PMID- 11003762 TI - Evaluation of hemostatic and coagulation factor abnormalities in patients undergoing the Fontan operation. AB - OBJECTIVE: Low-velocity and nonlaminar flow patterns in the Fontan circulation, as well as abnormal liver function in some patients, may partly account for the coagulation abnormalities seen. We examined (1) coagulation factor abnormalities before and after the Fontan procedure and (2) regional coagulation factor abnormalities in the Fontan circulation. METHODS: Levels of factors V, VII, VIII, X, antithrombin III, prothrombin fragment F1+2, protein C, and protein S were measured in 2 groups of patients: In 14 patients undergoing the Fontan procedure, blood was analyzed before the operation and 5 days after the operation (group 1). The median age in this group was 3.2 years. In 10 patients who had undergone the Fontan procedure, cardiac catheterization was performed and samples were taken from the femoral vein, inferior vena cava, right atrium, and pulmonary artery (group 2). The median age in this group was 6.2 years and the median follow-up from the Fontan procedure was 4.1 years. RESULTS: In group 1 a significant increase was noted postoperatively in the concentration of factor VIII (P<.001), factor X (P<.001), and prothrombin fraction F1+2 (P <.001). A significant decrease in the levels of antithrombin III (P <.001), protein C (P<.004), and protein S (P<.02) was also found. The increase in factors VIII and X persisted at 4 years' follow-up in group 2 patients. In group 2, no significant regional differences were observed between the coagulation factors measured at different sites. CONCLUSIONS: There is an increased tendency toward coagulation after the Fontan procedure. A prothrombotic state is supported by thrombin generation associated with reduced antithrombin III concentration. This increase in coagulation may contribute to the early and late risks of thromboembolism observed after the Fontan procedure. We did not find any regional differences in coagulation abnormalities in patients late after the Fontan procedure. Therefore, the mechanisms and causes of the coagulation abnormalities remain unclear. PMID- 11003763 TI - Preoperative autologous donation of blood for a simple cardiac anomaly: analysis of children weighing under twenty kilograms. AB - OBJECTIVE: Preoperative autologous donation of blood has been expanded to cardiac operations in children. However, because of problems such as lack of cooperation and hemodilution during cardiopulmonary bypass, its efficacy in small children is unclear. This study clarifies the clinical significance of preoperative autologous donation of blood in small children. METHODS: Thirty-seven patients weighing under 20 kg (age range, 3-9 years; weight range, 13-20 kg) underwent preoperative autologous donation and cardiac operations to treat a simple anomaly. Twenty-five age- and weight-matched patients who were not cooperative or refused preoperative autologous donation served as control subjects. Autologous blood was collected by the simple or leapfrog method and stored as blood components. Each collecting volume was 5 to 10 mL/kg. RESULTS: The donation was performed 6+/-2 times during 50+/-16 days, and the whole storage volume was 48+/ 17 mL/kg. There was no serious complication. The minimum hematocrit level negatively correlated with the priming volume of cardiopulmonary bypass (preoperative donation patients: P<.01, r(2) = 0.4; control subjects: P =.5, r (2) = 0.03). Blood loss did not significantly differ between preoperative donation patients and control subjects, and the transfused blood volumes were 43+/-13 mL/kg and 29+/-22 mL/kg, respectively. All of the autologous blood products but fresh frozen plasma were reinfused. Use of homologous blood was significantly less in preoperative donation patients than in control subjects (0% vs 80%, P <.01). In preoperative donation patients postoperative recovery in hemoglobin level was significantly better, which is concurrent with a higher reticulocyte level. CONCLUSION: Preoperative autologous donation can be performed safely with clinical efficacy, even in children under 20 kg. This can be improved further through coupling with another procedure. PMID- 11003764 TI - Effects of diltiazem prophylaxis on the incidence and clinical outcome of atrial arrhythmias after thoracic surgery. AB - OBJECTIVES: We sought to determine whether early prophylaxis with an L -type calcium channel blocker reduces the incidence and morbidity associated with atrial fibrillation/flutter and supraventricular tachyarrhythmia after major thoracic operations. METHODS: In this randomized, double-blind, placebo controlled study, 330 patients were given either intravenous diltiazem (n = 167) or placebo (n = 163) immediately after lobectomy (> or =60 years) or pneumonectomy (> or =18 years) and orally thereafter for 14 days. The primary end point with respect to efficacy was a sustained (> or =15 minutes) or clinically significant atrial arrhythmia during treatment. RESULTS: Postoperative atrial arrhythmias (atrial fibrillation/flutter = 60; supraventricular tachyarrhythmias = 5) occurred in 25 (15%) of the 167 patients in the diltiazem group and 40 (25%) of the 163 patients in the placebo group (P = .03). When compared with placebo, diltiazem nearly halved the incidence of clinically significant arrhythmias (17/167 [10%] vs. 31/163 [19%], P = .02). The 2 groups did not differ in the incidence of other major postoperative complications or overall duration or costs of hospitalization. No serious adverse effects caused by diltiazem were seen. CONCLUSIONS: After major thoracic operations, prophylactic diltiazem reduced the incidence of clinically significant atrial arrhythmias in patients considered at high risk for this complication. PMID- 11003765 TI - Results of surgical treatment of lung cancer involving the diaphragm. AB - OBJECTIVES: Lung cancers with diaphragmatic invasion are categorized as T3 lesions, but the surgical results have not been well known. We retrospectively surveyed patients with resected primary lung cancers involving the diaphragm. METHODS: A total of 16,771 patients underwent surgical resection for lung carcinoma between 1986 and 1995 at 31 institutions of the Lung Cancer Surgical Study Group in Japan. By investigating the database, we identified 63 patients (0. 38%) who underwent resection of T3 lung cancer invading the diaphragm. These patients constituted the study population, and their clinical and pathologic records were retrospectively analyzed. RESULTS: Tumor invasion to the diaphragm was diagnosed before operation only in 17 patients (27.0%). Complete resections of the primary lung tumors with the invaded diaphragm were performed in 55 patients (87.3%), of whom 26 had T3 N0 M0 diseases and 29 had T3 Nl-2 M0 diseases. The operative mortality was 1.6% in all patients. The 5-year survival of patients with complete resection was 22.6%, but there was no 4-year survivor in patients with incomplete resection (P =.024). The survivals of patients with completely resected T3 N0 M0 and T3 N1-2 M0 tumors were 28.3% and 18.1%, respectively (P =.013). In those patients, the depth of diaphragmatic involvement significantly affected the prognosis. The 5-year survival of the patients with shallow invasion (parietal pleura or subpleural tissue involvement) was 33.0%, whereas that of the patients with deep invasion (muscle or peritoneal infiltration) was 14.3% (P =.036). CONCLUSIONS: In selected patients with lung carcinoma and diaphragmatic invasion, combined resection of the lung and diaphragm offers the prospect of cure with acceptable mortality. However, primary lung tumors with diaphragmatic invasion, especially invasion of the muscle layer or deeper tissue, are not considered to be T3 lesions, because these cancers are generally technically resectable but oncologically almost incurable. PMID- 11003766 TI - Endoscopic coronary artery bypass grafting-the first steps on a long journey. PMID- 11003767 TI - Closed-chest coronary artery bypass grafting on the beating heart with the use of a computer-enhanced surgical robotic system. PMID- 11003768 TI - Closed-chest coronary artery surgery on the beating heart with the use of a robotic system. PMID- 11003769 TI - Three-dimensional computed tomographic angiography as preoperative evaluation of a patent internal thoracic artery graft. PMID- 11003770 TI - Occult active giant cell aortitis necessitating surgical repair. PMID- 11003771 TI - Phenoxybenzamine prevents spasm in radial artery conduits for coronary artery bypass grafting. PMID- 11003772 TI - Endoluminal replacement of the entire aorta for acute type A aortic dissection in a patient with Marfan syndrome. PMID- 11003773 TI - Double-barreled conduit for right atrioventricular connection in tricuspid atresia: a new technique. PMID- 11003774 TI - Mitral homograft replacement of tricuspid valve in children. PMID- 11003775 TI - Rapid enlargement of a distal arch aneurysm after endovascular stent-grafting for pseudoaneurysm: case report. PMID- 11003776 TI - Coronary artery bypass grafting without cardiopulmonary bypass in pheochromocytoma. PMID- 11003777 TI - Are adenosine myocardial protective effects mediated through cAMP-independent effects or through cAMP inhibition/production? PMID- 11003779 TI - S-100beta protein: yet uncertain role as a marker of cerebral injury in cardiac surgery. PMID- 11003780 TI - Genetic heterogeneity in hereditary sensory and autonomic neuropathies: The need for improved ascertainment. PMID- 11003781 TI - Molecular basis of muscular dystrophies. AB - Muscular dystrophies represent a heterogeneous group of disorders, which have been largely classified by clinical phenotype. In the last 10 years, identification of novel skeletal muscle genes including extracellular matrix, sarcolemmal, cytoskeletal, cytosolic, and nuclear membrane proteins has changed the phenotype-based classification and shed new light on the molecular pathogenesis of these disorders. A large number of genes involved in muscular dystrophy encode components of the dystrophin-glycoprotein complex (DGC) which normally links the intracellular cytoskeleton to the extracellular matrix. Mutations in components of this complex are thought to lead to loss of sarcolemmal integrity and render muscle fibers more susceptible to damage. Recent evidence suggests the involvement of vascular smooth muscle DGC in skeletal and cardiac muscle pathology in some forms of sarcoglycan-deficient limb-girdle muscular dystrophy. Intriguingly, two other forms of limb-girdle muscular dystrophy are possibly caused by perturbation of sarcolemma repair mechanisms. The complete clarification of these various pathways will lead to further insights into the pathogenesis of this heterogeneous group of muscle disorders. PMID- 11003782 TI - Electrophysiological features of inherited demyelinating neuropathies: A reappraisal in the era of molecular diagnosis. AB - The observation that inherited demyelinating neuropathies have uniform conduction slowing and that acquired disorders have nonuniform or multifocal slowing was made prior to the identification of mutations in myelin-specific genes which cause many of the inherited disorders involving peripheral nerve myelin. It is now clear that the electrophysiological aspects of these disorders are more complex than previously realized. Specifically, certain mutations appear to induce nonuniform slowing of conduction which resemble the findings in acquired demyelinating neuropathies. It is clinically important to recognize the different electrodiagnostic patterns of the various inherited demyelinating neuropathies. In addition, an understanding of the relationship between mutations of specific genes and their associated neurophysiological findings is likely to facilitate understanding of the role of these myelin proteins in peripheral nerve function and of how abnormalities in myelin proteins lead to neuropathy. We therefore review the current information on the electrophysiological features of the inherited demyelinating neuropathies in hopes of clarifying their electrodiagnostic features and to shed light on the physiological consequences of the different genetic mutations. PMID- 11003783 TI - Electrodiagnostic studies in amyotrophic lateral sclerosis and other motor neuron disorders. AB - The clinical electrodiagnostic medicine (EDX) consultant asked to assess patients with suspected amyotrophic lateral sclerosis (ALS) has a number of responsibilities. Among the most important is to provide a clinical assessment in conjunction with the EDX study. The seriousness of the diagnoses and their enormous personal and economic impact require a high-quality EDX study based on a thorough knowledge of and experience with motor neuron diseases (MNDs) and related disorders. Clinical evaluation will help determine which of the EDX tools available to the EDX consultant should be applied in individual patients. Although electromyography (EMG) and nerve conduction study are the most valuable, each of the following may be helpful in the assessment of selected patients based on their clinical findings: repetitive nerve stimulation, motor unit number estimate, single-fiber EMG, somatosensory evoked potential, autonomic function test, and polysomnography. The pertinent literature on these is reviewed in this monograph. The selection and application of these EDX tools depend on a thorough knowledge of the MNDs and related disorders. PMID- 11003784 TI - Ethical issues in DNA testing. AB - Molecular tests for single gene disorders have become the "gold standard," surpassing in sensitivity and specificity other ancillary tests such as imaging or neurophysiologic tests. Although direct technical problems are rare for DNA tests, few outcomes studies have been reported that examine sensitivity, specificity, and usage of tests outside the setting of academic medical centers. Ethical problems of DNA testing are mainly discussed in a theoretical framework and few published studies or case reports deal with real-life situations. Among neurologists, ethical dilemmas and attempts at their resolution are communicated mainly on a verbal basis, and even then relatively little follow-up is available. The potential use of information obtained through DNA testing by other family members, employers, insurance companies, and governmental agencies deserves scientific investigation and societal discussion. A major educational effort is needed to inform patients, providers, and third parties that the presence of a particular allele in an individual is only one piece in a complex puzzle of environmental and genetic interactions that may ultimately result in disease. PMID- 11003785 TI - Phenotypic and genetic study of a family with hereditary sensory neuropathy and prominent weakness. AB - We report the clinical and electrophysiological features of six members of a French family with a dominantly inherited motor and sensory neuropathy. Mean age at onset was 33.6 +/- 9.1 years. Mean age at examination was 55.5 +/- 13.3 years. Clinical presentation combined symptoms of hereditary sensory and autonomic neuropathy type I (HSAN-I) with prominent distal muscle weakness. Five male patients presented with sensory symptoms involving the distal part of the limbs, especially the legs. All but one had histories of trophic alterations, consisting of poorly healing foot ulcers. Muscle weakness and wasting were always present, often severe, and mainly affected dorsiflexion of the toes and feet. One obligate female carrier aged 65 was clinically asymptomatic. Electrophysiological findings were consistent with a distal axonal motor and sensory neuropathy. Results of linkage analysis excluded the Charcot-Marie-Tooth 2A (CMT2A) and CMT2B loci and suggested the possibility of a linkage to HSAN-I locus on 9q22.1-q22.3. PMID- 11003786 TI - Hydrogen peroxide decelerates recovery of action potential after high-frequency fatigue in skeletal muscle. AB - Effects of reactive oxygen species (ROS), especially hydrogen peroxide (H(2)O(2)), on recovery of action potential by resting for 30 min after high frequency fatigue were studied using frog skeletal muscle fibers. After stimulation at a frequency of 50 HZ for 2 min, the action potential amplitude was decreased by 14.5 mV from controls, and resting membrane was depolarized by 15.4 mV. Action potential duration was also prolonged by high-frequency stimulation (1.5 ms in controls to 2.6 ms). The high-frequency stimulation used here caused no muscle damage. The action potential was partially improved after a 30-min rest. Addition of catalase at 500 units/ml or H(2)O(2) at 0.5 mM to sartorius muscle did not alter any of the parameters of the action potential after high frequency stimulation. Treatment with catalase accelerated post-fatigue recovery of the action potential. Application of H(2)O(2) delayed post-fatigue recovery of resting and action potentials. When added to detubulated toe muscle fibers, catalase no longer improved the attenuation of action potential induced by high frequency stimulation, even after a 30-min rest. These findings suggest that removal of H(2)O(2) from transverse tubules is effective for post-fatigue recovery of action potential in skeletal muscle. PMID- 11003787 TI - Comparison of multiple point and statistical motor unit number estimation. AB - This study compares two common techniques for motor unit number estimation, multiple point stimulation and statistical method, to determine which is more reproducible. Surface recorded motor unit action potentials (SMUPs) of the left hypothenar muscle group were measured on 20 controls and 10 ALS patients. For multiple point, 10 different threshold SMUPs were recorded. For statistical method, mean SMUP amplitude was measured at several stimulus levels, typically spanning >40% of CMAP amplitude range. Both techniques were performed twice, results averaged, electrodes changed, and all recording repeated. For controls, mean of two motor unit number estimation (MUNE) (+/- standard deviation) was 60 (+/-5) for statistical method, and 108 (+/-38) for multiple point. For ALS patients, these values were 21 (+/-16) for statistical method and 55 (+/-39) for multiple point. Test-retest correlation coefficients and coefficients of variation for mean of two MUNE were 0.98 and 7% for statistical method, and 0.90 and 12% for multiple point, respectively. Statistical method was more reproducible and faster than multiple point, supporting its utility in monitoring rates of MUNE change. PMID- 11003788 TI - Contractile properties and fatigue of quadriceps muscles in multiple sclerosis. AB - Functional characteristics of electrically stimulated quadriceps muscles of patients with multiple sclerosis (MS) were determined to investigate whether adaptations in muscle properties contribute to the higher fatigability of these patients. The estimated maximal isometric force generating capacity of MS patients was only 11.2% (P < 0.05) lower than control subjects. However, the patients were only able to voluntarily exert 75 +/- 22% (n = 12) of their maximal capacity, against 94 +/- 6% (n = 7) for the control subjects. There were no differences in muscle speed, suggesting that muscle fiber distribution was not different in the MS patients due to reduced muscle usage. During a series of repeated contractions, greater decrements occurred in isometric force and in maximal rate of force rise in the MS patients (by 31.3 +/- 10.3% and 50.1 +/- 10.0%, respectively; n = 13) than control subjects (23.8 +/- 6.6% and 39.0 +/- 8.1%, n = 15), suggesting a lower oxidative capacity. The results indicate that increasing the mass of their muscles by training may help to reduce the excessive muscle fatigue of MS patients. PMID- 11003789 TI - Mapping motor cortex projections to single motor units in humans with transcranial magnetic stimulation. AB - We devised a method to investigate the cortical organization of corticomotoneurons (CMs) to upper limb muscles. A spike-triggering technique was used, in which a tonically discharging single motor unit (SMU) triggered transcranial magnetic stimulation (TMS) of motor cortex, and the probability of producing short-latency discharges (primary excitatory responses [PERs]) was measured. PER probabilities were mapped in 34 SMUs, using a 16 cm(2) scalp grid with the central reference point having a probability of 0.5. Maps showed a single optimum point of scalp stimulation and significant decreases in PER probability with shifts of 2 cm from this point, for all subjects. These findings suggest that the colony of CMs projecting to an individual SMN is contained within a small volume of motor cortex. Changes in PER probability with shifts in stimulation site may reflect the organization of other intracortical neurons mediating TMS activation of these CMs. PMID- 11003790 TI - Nuclear accumulation of expanded PABP2 gene product in oculopharyngeal muscular dystrophy. AB - Autosomal dominant oculopharyngeal muscular dystrophy (OPMD) is an adult-onset disease caused by (GCG) repeat expansions in exon 1 of the poly(A) binding protein 2 gene (PABP2). To elucidate the molecular mechanism underlying the disease, we raised an antiserum against a synthetic peptide fragment predicted from PABP2 cDNA. The peptide corresponded to amino acids 271-291 where a cluster of posttranslational arginine methylation occurs. We examined the subcellular localization of PABP2 in muscle specimens from five patients with OPMD, 14 patients with various neuromuscular disorders, and three normal controls. All Japanese patients with OPMD have been shown to have expanded (GCG)(8, 9, or 11) mutations in PABP2, as well as intranuclear tubulofilamentous inclusions (ITFI) of 8.5 nm. None of 50 separate Japanese control individuals were shown to have expanded (GCG) repeat in PABP2. Positive immunoreaction for polyclonal PABP2 was confined to the intranuclear aggregates of muscle fibers exclusively in patients with OPMD. Frequency of the nuclei positive for PABP2 (2%) was similar to that of ITFI detected by electron microscopy (2.5%). There was no apparent relationship between the frequency of PABP2-positive intranuclear aggregates and the severity of muscle fiber damage. In contrast, nuclear immunoreaction was not detected in any samples from normal controls or from other neuromuscular diseases. These results suggest the presence of molecular modification of the product of expanded (GCG) repeat in PABP2, since the synthetic antigen peptide may not recognize a highly dimethylated cluster of arginine residues of the native PABP2, but may recognize the mutated form. Nuclear accumulation of expanded PABP2 product implies a causative role for ITFI. PMID- 11003791 TI - Median nerve recovery in carpal tunnel syndrome. AB - We studied recovery of median nerve function in operated elderly and younger patients with carpal tunnel syndrome, compared to nonoperated patients, using neurophysiological methods. Three groups were analyzed separately: elderly operated (n = 16, aged 70 to 89); younger operated (n = 83, aged 30 to 69); and nonoperated (n = 52, aged 25 to 83). The elderly operated patients had significant improvement after surgery in all sensory nerve variables and a decrease in motor distal latency. The results were comparable to the results in the younger operated patients. In the nonoperated hands, the same neurophysiological variables were also significantly improved, but to a lesser extent than in the operated hands. Overall symptom relief was better in the operated hands. PMID- 11003792 TI - Metabolism of rat skeletal muscle after spinal cord transection. AB - We investigated the energy metabolism of the gastrocnemius muscle of the rat after spinal cord transection, using in vivo (31)P magnetic resonance spectroscopy (MRS). Spectra were obtained at rest and during exercise and recovery before, and at different time-points after, spinal cord transection. At rest, the adenosine triphosphate (ATP) level was not altered and the intracellular pH became permanently more alkaline. In electrically stimulated muscle, cord transection caused a greater phosphocreatine depletion than in control animals, and the maximum rate of oxidative ATP synthesis was significantly diminished; at days 30 and 60 after transection, an intracellular acidification was observed at the end of exercise. These effects indicate that, as in humans, spinal cord transection in rats leads to a decrease in mitochondrial oxidative metabolism and probably to an increase in anaerobic metabolism. This experimental model may prove useful for evaluating various approaches to improve muscle function in paraplegia. PMID- 11003793 TI - Clinical and pathologic features of focal myositis. AB - To clarify the nosology of focal myositis (FM), we report the clinical and pathologic features of eight patients presenting with focal enlargement of one muscle. Most patients improved without immunosuppressive therapy, and none developed polymyositis. Pathologic features were those of an inflammatory myopathy, with muscle fiber hypertrophy and moderate to severe inflammation. In most cases, a clustering of tightly packed muscle fibers, enveloped by a thick bundle of fibrosis, was associated with the diagnosis of FM. Immunohistochemistry showed T cell predominance within the interstitial infiltrates in all cases. No evidence of vasculitis was present. Our findings suggest that FM is a benign condition that has certain clinical features separating it from other inflammatory myopathies. Pathologic changes, such as large clusters of nesting muscle fibers surrounded by thick fibrosis, are more characteristic of FM than polymyositis. PMID- 11003794 TI - Effects of resistance training on neuromuscular junction morphology. AB - The aim of this study was to determine the impact of resistance exercise on neuromuscular junction (NMJ) architecture. Eighteen Sprague-Dawley rats either participated in a 7-week resistance training program or served as untrained controls. Following the experimental period, the NMJs of soleus muscles were visualized with immunofluorescent techniques, and muscle fibers were stained histochemically. Results indicate that resistance training significantly (P < 0.05) increased endplate perimeter length (15%) and area (16%), and significantly enhanced the dispersion of acetylcholine receptors within the endplate region. Pre- and post-synaptic modifications to resistance exercise were well-coupled. No significant alterations in muscle fiber size or fiber type were detected. The data presented here indicate that the stimulus of resistance training was sufficiently potent to remodel NMJ structure, and that this effect cannot be attributed to muscle fiber hypertrophy or fiber type conversion. PMID- 11003795 TI - Acquired slow-channel syndrome. AB - We report the case of a 37-year-old man with clinical and electrophysiological features of hereditary slow-channel syndrome (SCS) and antibodies against acetylcholine receptors (AChR-Abs). He presented with weakness of shoulder and hand muscles. A supramaximal single stimulus to the motor nerves disclosed a double compound muscle action potential (CMAP). Repetitive stimulation of ulnar, suprascapular, and median nerves showed a CMAP decrement greater than 10%. The patient responded to pyridostigmine. This report confirms the importance of AChR Ab titers in suspected cases of hereditary SCS because patients with positive AChR-Abs may have a better response to available treatments. PMID- 11003796 TI - Functional properties of regenerating skeletal muscle following LIF administration. AB - We tested the hypothesis that periodic systemic administration of the myogenic cytokine leukemia inhibitory factor (LIF) enhances the functional recovery of regenerating skeletal muscle following bupivacaine-induced degeneration. LIF had no effect on functional capacity or regenerating myofiber size in rat muscles at 7, 14, or 21 days post-injury. The results do not support exogenous administration of LIF as a treatment for acute muscle injury, but a more frequent dosing regimen should be tested. PMID- 11003797 TI - Estimation of cervical cord dysfunction by somatosensory evoked potentials. AB - The purpose of this study was to examine the relationship of abnormal short latency somatosensory evoked potentials (SSEPs) recorded by a noncephalic reference montage with clinical variables in cervical myelopathy patients and to reexamine the diagnostic utility of SSEPs in such patients. We studied cervical SSEPs elicited by stimulating the median and ulnar nerves in 87 patients. Our grade classification of spinal N13, which is based on the normal limits of latencies or amplitudes, corresponded well with the clinical variables and is of value when trying to localize the cervical lesion segmentally. The N9-P14 interpeak latency in response to ulnar nerve stimulation correlated well with lower extremity function (r = -0.440, P <0.0001). We suggest a combined assessment of N13 amplitude, and N9-N13 and N9-P14 interpeak latencies to estimate dorsal column and dorsal horn function separately in patients with cervical myelopathy. PMID- 11003798 TI - Reproducibility of intracortical inhibition and facilitation using the paired pulse paradigm. AB - We have evaluated the reproducibility of intracortical inhibition (ICI) and facilitation (ICF) studied with paired-pulse focal transcranial magnetic stimulation. Three investigators studied the same subjects (n = 4) in three different sessions. A high variability was shown across subjects [coefficient of variation, (cv) 67.3% for ICI and 21.2% for ICF]. Intersession variability was up to 37.1% for ICI and 22.7% for ICF. Interinvestigator variability was 17.3% for ICI and negligible for ICF. Our results may have implications for planning future studies. PMID- 11003799 TI - Mild ciguatera poisoning: Case reports with neurophysiological evaluations. AB - Ciguatera poisoning causes mainly gastrointestinal and neurological effects of variable severity. However, symptoms of peripheral neuropathy with paresthesias and paradoxical disturbance of thermal sensation are the hallmark. Electrophysiological studies are often normal, except in severe cases. We report four people who developed mild ciguatera poisoning after barracuda ingestion. Electrophysiological studies documented normocalcemic latent tetany. These findings are consistent with ciguatoxin's mechanism of toxicity, which involves inactivation of voltage-gated Na(+) channels and eventually increases nerve membrane excitability. PMID- 11003800 TI - Clinicopathological and molecular biological studies in a patient with neurolymphomatosis. AB - We describe a patient with a clinical disorder that resembled vasculitic neuropathy in which peripheral nerves were successively affected over several months, but without systemic involvement. An initial muscle biopsy near the involved nerves showed signs of nonspecific inflammation around the muscle and nerve fibers. Immunosuppressive treatment resulted in a dramatic reduction in pain, but relapses of the disease eventually occurred, and the patient died 22 months after onset of the first symptoms. Pathologically, a malignant non Hodgkin's B-cell lymphoma, restricted to the intra- and extradural peripheral nervous system, was found. The demonstration by Southern blotting of immunoglobulin heavy chain gene rearrangement confirmed the monoclonal nature of the lymphomatous cells. In situ hybridization tests for Epstein-Barr and herpes virus subtypes were negative. Our case underlines i) how difficult diagnosis can be despite extensive investigations, ii) the usefulness of immunosuppressive treatment in the early stage of the disease, iii) the importance of immunostaining and genome analysis for distinguishing between different types of human neurolymphomatosis, and iv) the fact that the initial inflammatory process in the muscle biopsy may be interpreted either as a paraneoplastic effect of the lymphoma or as a viral inflammatory neuromyopathy that triggers the development of the malignant lymphoma. PMID- 11003801 TI - Monofocal motor neuropathy responsive to intravenous immunoglobulins. PMID- 11003803 TI - The anti-reflux barrier and mechanisms of gastro-oesophageal reflux. AB - The lower oesophageal sphincter (LOS) is the major component of the anti-reflux barrier. The majority of reflux episodes occur because of intermittent brief complete lower oesophageal sphincter relaxations, transient LOS relaxations, rather than from chronic absence of LOS pressure. Recent advances in the understanding of the neural mechanisms and the receptors involved in the triggering of transient LOS relaxations have provided new insights into their control and offer the potential for the development of pharmacological therapy of reflux based on control of these events. Extrinsic support by the crural diaphragm is also important and loss of this support through development of hiatus hernia significantly compromises LOS function. This chapter reviews the components of the anti-reflux barrier, the patterns and mechanisms of LOS dysfunction underlying reflux episodes, and the interplay between sphincteric and non-sphincteric factors. PMID- 11003804 TI - Mechanisms of oesophageal mucosal defence. AB - The integrity of the oesophageal mucosa depends upon an equilibrium between aggressive factors, predominantly acid and pepsin, and protective mechanisms. Protective mechanisms operate within the oesophageal mucosa as pre-epithelial, epithelial and post-epithelial defences. Only the protective components of the oesophageal pre-epithelial defence can be tested in vivo in humans. It has been recently demonstrated that human oesophageal submucosal glands elaborate mucous secretion rich in bicarbonate and non-bicarbonate buffers, mucin, prostaglandin E(2), epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha). This oesophageal secretion, accompanied by similarly protective factors within the swallowed saliva, defines the protective potential of the oesophageal pre-epithelial defence that exists in the form of a mucus-buffer layer covering the oesophageal mucosa and which retards the back-diffusion of hydrogen ions. It has also been demonstrated that patients with severe erosive reflux oesophagitis exhibit qualitative impairment in both the salivary and oesophageal components of the oesophageal pre-epithelial defence. Furthermore, patients with endoscopically negative gastro-oesophageal reflux disease have a significantly stronger oesophageal pre-epithelial defence than patients with erosive reflux oesophagitis. On the other hand, African-Americans, who are less likely to develop erosive reflux oesophagitis, have a stronger oesophageal pre epithelial defence than do Caucasians. The salivary component of the oesophageal pre-epithelial defence can be enhanced by mastication and the administration of cisapride, whereas oesophageal secretion can only be significantly augmented by cisapride. PMID- 11003805 TI - Duodenogastro-oesophageal reflux. AB - The role of duodenogastro-oesophageal reflux (DGOR), once erroneously termed 'bile reflux', in causing oesophageal mucosal damage has been an area of interest in both animal and human studies. However, due to the lack of appropriate techniques for accurately measuring DGOR, extrapolation of findings from animal studies to humans was difficult to make. The recent advent of the Bilitec system, an ambulatory bilirubin monitoring device, is increasing our knowledge of the specific role of DGOR in oesophageal diseases. Studies suggest that the DGOR without acid reflux may result in symptoms but unless acid reflux is present simultaneously, it does not cause oesophagitis. Therefore, our therapies should aim at reducing both DGOR and acid reflux. Studies show that this may be accomplished by anti-reflux surgery or the use of proton pump inhibitors, which, by reducing gastric volume, decrease the damaging potential of both acid and DGOR. PMID- 11003806 TI - Helicobacter pylori infection and gastro-oesophageal reflux disease: coincidence or association? AB - Concerning the relationship between Helicobacter pylori infection and gastro oesophageal reflux disease (GORD), the debate is ongoing whether the infection confers protection, is harmful or whether both entities are independent. Epidemiological evidence is given for an increased prevalence of GORD and a decreased prevalence of H. pylori infection in the western world. The assumpton derived from it is that H. pylori protects from GORD. Pathophysiological aspects need to consider the type and expression of gastritis which is associated with varying changes of gastric function. Depending on the type of gastritis, acid secretion may either increase or decrease and thereby impact on acid exposure of the oesophagus. Other changes related to the role of H. pylori in pathophysiology of GORD are still hypothetical. Clinical data are controversial whether or not GORD increases after H. pylori eradication. Prospective studies including characterization of strains and gastric physiology will clarify this issue. An accelerated induction of gastric mucosal atrophy in patients on long-term proton pump inhibitors is reported in most available studies. An increase of inflammatory activity in fundic and corpus mucosa is a consistent phenomenon. Therefore, in the authors' opinion, eradication appears advisable. PMID- 11003807 TI - Epidemiology and natural history of reflux disease. AB - Gastro-oesophageal reflux disease (GORD) is a highly prevalent condition in Western countries; at least 20% of the population have weekly symptoms. The incidence appears to be rising in the West and in some developing countries. Heartburn, based on a carefully elicited history, is reasonably specific for identifying GORD if it is a predominant complaint. Symptoms, however, appear to correlate poorly with oesophagitis; hence, severe symptoms do not indicate there is greater oesophageal damage. Only one-third to one-half of patients with GORD undergoing endoscopy have oesophagitis. GORD is usually a chronic disease but one third may lose their symptoms over time. An ill-defined subset of patients over time may progress to develop abnormal acid exposure or oesophagitis, or both, when none existed at baseline. GORD has a significant negative impact on quality of life to the same degree as other chronic medical conditions, but impairment in quality of life is independent of oesophagitis. PMID- 11003808 TI - Diagnosis of reflux disease. AB - There are numerous tests for which a diagnostic value in the context of gastro oesophageal reflux disease has been claimed. Some of these tests (e.g. the acid perfusion test) have become obsolete after the advent of 24-hour oesophageal pH monitoring. With the latter test not only can excessive reflux be identified, but also, and more importantly, a temporal relationship can be demonstrated between a patient's symptoms and reflux episodes. Radiographical examination of the oesophagus has largely been replaced by endoscopy, although the use of the former test is still indicated in certain circumstances (e.g. in the differentiation of sliding from para-oesophageal hiatus hernia). In clinical practice, the so-called proton pump inhibitor test has gained considerable popularity. Despite several studies on the specificity and sensitivity of this test, its value has not yet been established with sufficient accuracy. Conventional manometric evaluation of lower oesophageal sphincter pressure has been over-emphasized as a diagnostic test in gastro-oesophageal reflux disease. PMID- 11003809 TI - Strategies for medical management of reflux disease. AB - Management of gastro-oesophageal reflux disease (GORD) patients must consider two issues: (i) how to optimize the treatment of a presenting symptom complex, and (ii) how to manage risk of adenocarcinoma associated with GORD. In most cases the need for, and potency of, pharmacological therapy used is decided by symptom assessment. Considering cost effectiveness, the three increments of pharmacological therapy are: (i) generic histamine(2)receptor antagonists, (ii) standard dose proton pump inhibitors, and (iii) higher dose proton pump inhibitors. Endoscopy is warranted if there is doubt regarding the diagnosis of GORD or if the patient relays alarm symptoms suggesting more ominous diagnoses (dysphagia, bleeding, weight loss, odynophagia). The other major indication for endoscopy is to screen for adenocarcinoma or Barrett's metaplasia in the patient with chronic symptoms. In most patients, the need for maintenance medical therapy is determined by the rapidity of symptom recurrence during a trial period off the medication. PMID- 11003810 TI - Anti-reflux surgery in the laparoscopic era. AB - The introduction of laparoscopic anti-reflux surgery has led to a renewed interest in the operative treatment of gastro-oesophageal reflux disease (GORD). Three groups of patients can be identified who are particularly suited to laparoscopic anti-reflux surgery. Failure to respond to medical treatment has been historically the main determinant for those referred for anti-reflux surgery. With the availability of modern anti-secretory drugs most patients with chronic GORD can control their symptoms adequately by these means. Even effective medical therapy, however, is not without problems. In many patients rapid and consistent relapse of symptoms and oesophagitis occurs on cessation of therapy. Some of these patients do not want to be reliant on a form of medication that has yet to firmly establish its record for safety over many years of continued use. A second readily identifiable group of patients are those who are often described as 'volume refluxers'. They are bothered by persistent fluid regurgitation despite adequate control of their heartburn with acid suppressive drugs. Third there are those individuals who develop oesophageal strictures and those with Barrett's oesophagus and concomitant reflux symptoms and also those with respiratory complications associated with presumed aspiration of gastric juice into the pharynx and into the respiratory tree. The low morbidity associated with laparoscopic surgery that has been achieved in the best modern series means that the pendulum may swing back to surgery and therefore it is even more important that the right operation (fundoplication) is done for the right patient. Failure to create an adequate crural repair behind the wrap is associated with a risk of early post-operative para-oesophageal herniation and proximal wrap migration. The question of tailored anti-reflux surgery based on the pre-operative motor function of the body of the oesophagus is widely applied, although the scientific basis for these selective approaches is rather weak. Partial fundoplication seems to be associated with very low rates of dysphagia and of gas bloat. Assessment of the post-operative result should ideally be done by an independent observer and should consider not only traditional outcome measures but also the impact of surgery on the quality of the patient's life. Investigations on the cost effectiveness of these surgical therapeutic strategies suggest important benefits of surgery, which should be incorporated into the clinical decision process when assessing different long-term management alternatives for patients with chronic GORD. PMID- 11003811 TI - Controversies in long-term management of reflux disease. AB - There are significant controversies about the long-term therapy of reflux disease that arise primarily from a lack of data on the risks of the disease and its therapies over the relevant time scale of several decades. Currently, there are no appropriately structured direct comparisons between the two current major treatment options of tailored long-term acid suppression and laparoscopic anti reflux surgery. Critical review of the available literature does not support the assertion that anti-reflux surgery has a superior risk/benefit profile when compared to long-term proton pump inhibitor (PPI) therapy, since non-controlled data indicate that although both therapies are relatively safe, morbidity and mortality rates are lowest with PPI therapy, whilst efficacy is comparable. The clinical significance of the reflux that continues to occur after both anti reflux surgery and during PPI therapy is uncertain, but probably over-estimated. As pathological duodenogastro-oesophageal reflux affects only a small minority of reflux disease patients, issues arising from it should not be regarded as mainstream influences on the choice of long-term therapy. Notwithstanding, this type of reflux is substantially reduced by both PPI therapy and anti-reflux surgery. The choice between laparoscopic surgery and long-term tailored acid suppression should be determined primarily by assessment of operative risk, the quality of surgery available to the patient and by patient preference, after balanced explanation of the risks and benefits of each option. Given that the cost of PPI therapy is likely to drop substantially in the next few years, drug cost should not be a major pressure for the choice of anti-reflux surgery. PMID- 11003812 TI - Endoscopy-negative reflux disease. AB - The majority of patients with symptoms of gastro-oesophageal reflux have no endoscopic evidence of oesophagitis. There has been remarkably little systematic gathering of information about this group of patients. It is commonly believed that they have a mild form of reflux disease, with low levels of dysfunction that usually respond to simple therapeutic measures. Emerging data from recent studies indicate that this is not the case. Endoscopy-negative patients have symptom severities comparable to those with erosive disease, and which significantly impair their quality of life. The limited data available on the pathophysiology of endoscopy-negative reflux disease suggest that, in the majority of patients, it is as much a disease of excessive gastro-oesophageal reflux as it is in patients with oesophageal lesions. The same principles that apply to successful treatment in patients with oesophagitis also hold true for patients with endoscopy-negative disease. PMID- 11003813 TI - Reflux in children. AB - Gastro-oesophageal reflux (GOR) is an extremely common paediatric problem that often runs a harmless and self-limited course. Physiological GOR however can lead to marked parental anxiety, many unnecessary investigations and often unwarranted and potentially harmful therapeutic interventions. Our ability to better define GOR and gastro-oesophageal reflux disease (GORD) has improved in the past 15 years with a better understanding of the pathophysiology in infants and children due to the development and wider use of flexible endoscopy, 24-hour oesophageal pH monitoring and, more recently, the use of micromanometric methods for studying oesophageal motility. This will be further enhanced in the future with the development of non-invasive breath testing to study gastrointestinal motility and the use of electrical impedance to study fluid movement. Our therapeutic interventions have also improved particularly in the areas of acid suppression, improved surgical techniques and most recently laparoscopic fundoplication. This chapter reviews these advances in the paediatric area especially with regard to pathophysiology, diagnostic testing and therapeutic intervention. PMID- 11003815 TI - Index PMID- 11003814 TI - Barrett's oesophagus: diagnosis and management. AB - Gastro-oesophageal reflux disease and its sequela, Barrett's oesophagus, are the major recognized risk factors for oesophageal adenocarcinoma, a tumour whose frequency has increased dramatically in Western countries over the past few decades. Barrett's oesophagus develops through the process of metaplasia in which one adult cell type replaces another. The metaplastic, intestinal-type cells of Barrett's oesophagus are predisposed to develop genetic changes that eventuate in cancer. This report reviews the recent controversy regarding diagnostic criteria for Barrett's oesophagus, and provides practical guidelines for identifying the condition. The risks and benefits of the proposed medical, surgical and endoscopic therapies for Barrett's oesophagus are discussed in detail, and the approach to management recently endorsed by the American College of Gastroenterology is summarized. PMID- 11003816 TI - Preface PMID- 11003817 TI - What it takes to fly: the structural and functional respiratory refinements in birds and bats. AB - In absolute terms, flight is a highly energetically expensive form of locomotion. However, with respect to its cost per unit distance covered, powered flight is a very efficient mode of transport. Birds and bats are the only extant vertebrate taxa that have achieved flight. Phylogenetically different, they independently accomplished this elite mode of locomotion by employing diverse adaptive schemes and strategies. Integration of functional and structural parameters, a transaction that resulted in certain trade-offs and compromises, was used to overcome exacting constraints. Unique morphological, physiological and biochemical properties were initiated and refined to enhance the uptake, transfer and utilization of oxygen for high aerobic capacities. In bats, exquisite pulmonary structural parameters were combined with optimal haematological ones: a thin blood-gas barrier, a large pulmonary capillary blood volume and a remarkably extensive alveolar surface area in certain species developed in a remarkably large lung. These factors were augmented by, for example, exceptionally high venous haematocrits and haemoglobin concentrations. In birds, a particularly large respiratory surface area and a remarkably thin blood-gas (tissue) barrier developed in a small, rigid lung; a highly efficient cross-current system was fabricated within the parabronchi. The development of flight in only four animal taxa (among all the animal groups that have ever evolved; i.e. insects, the now extinct pterosaurs, birds and bats) provides evidence for the enormous biophysical and energetic constraints that have stymied volancy. Bats improved a fundamentally mammalian lung to procure the large amounts of oxygen needed for flight. The lung/air sac system of birds is not therefore a prescriptive morphology for flight: the essence of its design can be found in the evolution of the reptilian lung, the immediate progenitor stock from which birds arose. The attainment of flight is a classic paradigm of the remarkable adaptability inherent in organismal and organic biology for countering selective pressures by initiating elegant morphologies and physiologies. PMID- 11003818 TI - Immunolocalization of Dpfp1, a byssal protein of the zebra mussel Dreissena polymorpha. AB - The zebra mussel is one of only a few freshwater bivalves known to produce a byssus. This fibrous, proteinaceous and highly cross-linked structure allows the mussel to attach to a variety of substrata and contributes to its notoriety as a major freshwater biofouling species. We have successfully expressed a full-length version of Dreissena polymorpha foot protein 1 (Dpfp1), a putative byssal thread precursor, and have used the recombinant protein as an antigen for polyclonal antibody production. Antisera obtained from rabbits immunized with recombinant Dpfp1 recognize the protein in western blots of extracts from foot tissue and byssal threads. On the basis of this evidence, we conclude that Dpfp1 is a byssal precursor protein manufactured and stored in the foot of the mussel. Immunohistochemical localization of Dpfp1 suggests that the protein is localized in secretory granules in a large gland surrounding the ventral groove of the foot. Only a subset of these glandular cells stockpiles the protein, implying that the zebra mussel foot is a complex organ capable of several distinct secretory activities involved in byssal thread formation. The uniform distribution of Dpfp1-containing cells suggests that the protein is a significant load-bearing component of zebra mussel byssal threads, although a more rigorous test of this hypothesis awaits ultrastructural localization of the protein in mature byssal threads. PMID- 11003819 TI - Ontogenetic changes in characteristics required for endothermy in juvenile black skipjack tuna (Euthynnus lineatus). AB - To characterize better the development of endothermy in tunas, we assessed how the abilities to generate heat and to conserve heat within the aerobic, slow twitch (red) myotomal muscle using counter-current heat exchangers (retia) change with size in juvenile black skipjack tuna (Euthynnus lineatus) above and below the hypothesized minimum size for endothermy of 207 mm fork length (FL). Early juvenile scombrids (10-77 mm FL) collected off the Pacific coast of Panama were raised to larger sizes at the Inter-American Tropical Tuna Commission Laboratory at Achotines Bay, Panama. Evidence of central and lateral rete blood vessels was found in E. lineatus as small as 95.9 mm FL and 125 mm FL, respectively. In larger E. lineatus juveniles (up to 244 mm FL), the capacity for heat exchange increased with fork length as a result of increases in rete length, rete width and the number of vessel rows. The amount (g) of red muscle increased exponentially with fork length in both E. lineatus (105-255 mm FL) and a closely related ectothermic species, the sierra Spanish mackerel Scomberomorus sierra (151-212 mm FL), but was greater in E. lineatus at a given fork length. The specific activity (international units g(-)(1)) of the enzyme citrate synthase in red muscle, an index of tissue heat production potential, increased slightly with fork length in juvenile E. lineatus (84. 1-180 mm FL) and S. sierra (122-215 mm FL). Thus, total red muscle heat production capacity (red muscle citrate synthase activity per gram times red muscle mass in grams) increased with fork length, primarily because of the increase in red muscle mass. Below 95.9 mm FL, E. lineatus cannot maintain red muscle temperature (T(m)) above the ambient water temperature (T(a)) because juveniles of this size lack retia. Above 95.9 mm FL, the relationship between T(x) (T(m)-T(a)) and FL for E. lineatus diverges from that for the ectothermic S. sierra because of increases in the capacities for both heat production and heat retention that result in the development of endothermy. PMID- 11003820 TI - Maximum sustainable speeds and cost of swimming in juvenile kawakawa tuna (Euthynnus affinis) and chub mackerel (Scomber japonicus). AB - Tunas (Scombridae) have been assumed to be among the fastest and most efficient swimmers because they elevate the temperature of the slow-twitch, aerobic locomotor muscle above the ambient water temperature (endothermy) and because of their streamlined body shape and use of the thunniform locomotor mode. The purpose of this study was to test the hypothesis that juvenile tunas swim both faster and more efficiently than their ectothermic relatives. The maximum sustainable swimming speed (U(max), the maximum speed attained while using a steady, continuous gait powered by the aerobic myotomal muscle) and the net cost of transport (COT(net)) were compared at 24 degrees C in similar-sized (116-255 mm fork length) juvenile scombrids, an endothermic tuna, the kawakawa (Euthynnus affinis) and the ectothermic chub mackerel (Scomber japonicus). U(max) and COT(net) were measured by forcing individual fish to swim in a temperature controlled, variable-speed swimming tunnel respirometer. There were no significant interspecific differences in the relationship between U(max) and body mass or fork length or in the relationship between COT(net) and body mass or fork length. Muscle temperatures were elevated by 1.0-2.3 degrees C and 0.1-0.6 degrees C above water temperature in the kawakawa and chub mackerel, respectively. The juvenile kawakawa had significantly higher standard metabolic rates than the chub mackerel, because the total rate of oxygen consumption at a given swimming speed was higher in the kawakawa when the effects of fish size were accounted for. Thus, juvenile kawakawa are not capable of higher sustainable swimming speeds and are not more efficient swimmers than juvenile chub mackerel. PMID- 11003821 TI - Swimming kinematics of juvenile kawakawa tuna (Euthynnus affinis) and chub mackerel (Scomber japonicus). AB - The swimming kinematics of two active pelagic fishes from the family Scombridae were compared to test the hypothesis that the kawakawa tuna (Euthynnus affinis) uses the thunniform mode of locomotion, in which the body is held more rigid and undergoes less lateral movement in comparison with the chub mackerel (Scomber japonicus), which uses the carangiform swimming mode. This study, the first quantitative kinematic comparison of size-matched scombrids, confirmed significantly different swimming kinematics in the two species. Ten kawakawa (15.1-25.5 cm fork length, FL) and eight chub mackerel (14.0-23.4 cm FL), all juveniles, were videotaped at 120 Hz while swimming at several speeds up to their maximum sustained speed at 24 degrees C. Computerized motion analysis was used to digitize specific points on the body in sequential video frames, and kinematic variables were quantified from the progression of the points over time. At a given speed, kawakawa displayed a significantly greater tailbeat frequency, but lower stride length, tailbeat amplitude and propulsive wavelength, than chub mackerel when size effects were accounted for. Midline curvatures subdivided on the basis of X-rays into individual vertebral elements were used to quantify axial bending in a subset of the fish studied. Maximum intervertebral lateral displacement and intervertebral flexion angles were significantly lower along most of the body in kawakawa than in chub mackerel, indicating that the kawakawa undergoes less axial flexion than does the chub mackerel, resulting in lower tailbeat amplitudes. However, lateral movement at the tip of the snout, or yaw, did not differ significantly interspecifically. Despite these differences, the net cost of transport was the same in the two species, and the total cost was higher in the kawakawa, indicating that the tuna juveniles are not more efficient swimmers. PMID- 11003822 TI - Effects of feeding on arterial blood gases in the American alligator Alligator mississippiensis. AB - Reptiles habitually ingest large meals at infrequent intervals, leading to changes in acid-base status as the net secretion of acid to the stomach causes a metabolic alkalosis (the alkaline tide). In chronically cannulated and undisturbed amphibians and reptiles, the pH changes in arterial blood are, nevertheless, reduced by a concomitant respiratory acidosis (increased P(CO2) caused by a relative hypoventilation). Alligators (Alligator mississippiensis) have been reported to exhibit exceptionally large increases in plasma [HCO3(-)] following feeding, but these studies were based on blood samples obtained by cardiac puncture, so stress and disturbance may have affected the blood gas levels. Furthermore, crocodilian haemoglobin is characterised by a unique binding of HCO3(-) that act to reduce blood oxygen-affinity, and it has been proposed that this feature safeguards oxygen offloading by counteracting pH effects on blood oxygen-affinity. Therefore, to study acid-base regulation and the interaction between the alkaline tide and oxygen transport in more detail, we describe the arterial blood gas composition of chronically cannulated and undisturbed alligators before and after voluntary feeding (meal size 7.5+/-1% of body mass). Digestion was associated with an approximately fourfold increase in metabolic rate (from 0.63+/-0.04 to 2.32+/-0.24 ml O(2) min(-1)kg(-1)) and was accompanied by a small increase in the respiratory gas exchange ratio. The arterial P(O2) of fasting alligators was 60.3+/-6.8 mmHg (1 mmHg = 0.133 kPa) and reached a maximum of 81.3+/-2.7 mmHg at 96 h following feeding; there was only a small increase in lactate levels, so the increased metabolic rate seems to be entirely aerobic. Plasma [HCO3(-)] increased from 24.4+/-1.1 to 36.9+/-1.7 mmol l(-1) (at 24 h), but since arterial P(CO2) increased from 29.0+/-1.1 to 36.8+/ 1.3 mmHg, arterial pH remained virtually unaffected (changing from 7.51+/-0.01 to 7.58+/-0.01 at 24 h). The changes in plasma [HCO3(-)] were mirrored by equimolar reductions in plasma [Cl(-)]. The in vitro blood oxygen-affinity was reduced during the post-prandial period, whereas the estimated in vivo blood oxygen affinity remained virtually constant. This supports the view that the specific HCO3(-) effect prevents an increased blood oxygen-affinity during digestion in alligators. PMID- 11003823 TI - Aerodynamic characteristics of dragonfly wing sections compared with technical aerofoils. AB - During gliding, dragonfly wings can be interpreted as acting as ultra-light aerofoils which, for static reasons, have a well-defined cross-sectional corrugation. This corrugation forms profile valleys in which rotating vortices develop. The cross-sectional configuration varies greatly along the longitudinal axis of the wing. This produces different local aerodynamic characteristics. Analyses of the C(L)/C(D) characteristics, where C(L) and C(D) are the lift and drag coefficients, respectively (at Reynolds numbers Re of 7880 and 10 000), using a force balance system, have shown that all cross-sectional geometries have very low drag coefficients (C(D, min)<0.06) closely resembling those of flat plates. However, the wing profiles, depending upon their position along the span length, attain much higher lift values than flat plates. The orientation of the leading edge does not play an important role. The detectable lift forces can be compared with those of technical wing profiles for low Re numbers. Pressure measurements (at Re=9300) show that, because of rotating vortices along the chord length, not only is the effective profile form changed, but the pressure relationship on the profile is also changed. Irrespective of the side of the profile, negative pressure is produced in the profile valleys, and net negative pressure on the upper side of the profile is reached only at angles of attack greater than 0 degrees. These results demonstrate the importance of careful geometrical synchronisation as an answer to the static and aerodynamic demands placed upon the ultra-light aerofoils of a dragonfly. PMID- 11003824 TI - Sanderlings (Calidris alba) have a magnetic compass: orientation experiments during spring migration in Iceland. AB - The migratory orientation of sanderlings (Calidris alba) was investigated with cage experiments during the spring migration in southwest Iceland. Sanderlings were exposed to 90 degrees counterclockwise-shifted magnetic fields under both clear skies and natural overcast. Clear sky control tests resulted in a northerly mean direction, in agreement with predictions based on ringing recovery data and earlier visual observations of departing flocks. Sanderlings closely followed experimental deflections of magnetic fields when tested under clear skies. Control experiments under natural overcast resulted in a bimodal distribution approximately coinciding with the magnetic north-south axis. Overcast tests did not reveal any predictable response to the experimental treatment, but instead resulted in a non-significant circular distribution. The time of orientation experiments in relation to the tidal cycle affects the motivation of the birds to depart, as shown by the lower directional scatter of headings of individuals tested within the appropriate tidal window under clear skies. Sanderlings were significantly more likely to become inactive under overcast conditions than under clear sky conditions. The results demonstrate, for the first time, that a wader species such as the sanderling possesses a magnetic compass and suggest that magnetic cues are of primary directional importance. However, overcast experiments indicate that both celestial and geomagnetic information are needed for sanderlings to realize a seasonally appropriate migratory orientation. PMID- 11003825 TI - Sustaining olfaction at low salinities: mapping ion flux associated with the olfactory sensilla of the blue crab Callinectes sapidus. AB - To test the hypothesis of a diffusion-generated, ionic/osmotic microenvironment within the olfactory sensilla (aesthetascs), flux gradients of Ca(2+) and K(+) associated with the external surfaces of these sensilla were spatially mapped using self-referencing, ion-selective microelectrodes. Blue crabs (Callinectes sapidus) acclimated to low-salinity conditions (15% sea water and fresh water) showed a net efflux of ions from the aesthetascs. The region of maximum flux associated with each aesthetasc conformed to that predicted from structural data and corresponded to the permeable region of the cuticle separating the olfactory dendrites from the external environment. Estimates of net flux from the entire tuft of aesthetascs for both Ca(2+) and K(+) fell within the predicted range on the basis of comparisons with (22)Na(+) flux measured previously and assuming a passive diffusion model of ion movement from the hemolymph to the sensillar lymph and, ultimately, to the external environment. The maximum concentrations of these ions measured deep within the tuft are discussed in the light of a potential across the aesthetascs that may limit ion efflux at low salinities. PMID- 11003826 TI - Flexibility in basal metabolic rate and evaporative water loss among hoopoe larks exposed to different environmental temperatures. AB - The 'energy demand' hypothesis for short-term adjustments in basal metabolic rate (BMR) posits that birds adjust the size of their internal organs relative to food intake, a correlate of energy demand. We tested this hypothesis on hoopoe larks (Alaemon alaudipes), inhabitants of the Arabian desert, by acclimating birds for 3 weeks at 15 degrees C and at 36 degrees C, then measuring their BMR and total evaporative water loss (TEWL). Thereafter, we determined the dry masses of their brain, heart, liver, kidney, stomach, intestine and muscles of the pectoral region. Although mean body mass did not differ initially between the two groups, after 3 weeks, birds in the 15 degrees C group had gained mass (44.1+/-6.5 g), whereas larks in the 36 degrees C group had maintained a constant mass (36.6+/ 3.6 g; means +/- s.d., N=6). Birds in the 15 degrees C group had a mean BMR of 46.8+/-6.9 kJ day(-1), whereas birds in the 36 degrees C group had a BMR of 32.9+/-6.3 kJ day(-1), values that were significantly different when we controlled for differences in body mass. When measured at 35 degrees C, larks in the cold-exposure group had a TEWL of 3.55+/-0.60 g H(2)O day(-)(1), whereas TEWL for birds in the 36 degrees C group averaged 2.23+/-0.28 g H(2)O day(-1), a difference of 59.2%. Mass-independent TEWL differed significantly between groups. Larks in the 15 degrees C group had a significantly larger liver, kidney and intestine than larks in the 36 degrees C group. The total increase in organ mass contributed 14.3% towards the total mass increment in the cold exposure group. Increased food intake among larks in the cold group apparently resulted in enlargement of some of the internal organs, and the increase in mass of these organs required a higher rate of oxygen uptake to support them. As oxygen demands increased, larks apparently lost more evaporative water, but the relationship between increases in BMR and TEWL remains unresolved. PMID- 11003827 TI - Crushing motor patterns in drum (Teleostei: Sciaenidae): functional novelties associated with molluscivory. AB - This study explores the evolution of molluscivory in the marine teleost family Sciaenidae by comparing the motor activity patterns of the pharyngeal muscles of two closely related taxa, the molluscivorous black drum (Pogonias cromis) and the generalist red drum (Sciaenops ocellatus). Muscle activity patterns were recorded simultaneously from eight pharyngeal muscles. Electromyographic (EMG) activity was recorded during feeding on three prey types that varied in shell hardness. Canonical variate and discriminant function analyses were used to describe the distinctness of drum pharyngeal processing behaviors. Discriminant functions built of EMG timing variables were more accurate than muscle activity intensity at identifying cycles by prey type and species. Both drum species demonstrated the ability to modulate pharyngeal motor patterns in response to prey hardness. The mean motor patterns and the canonical variate space of crushing behavior indicated that black drum employed a novel motor pattern during molluscivory. The mollusc-crushing motor pattern of black drum is different from other neoteleost pharyngeal behaviors in lacking upper jaw retraction by the retractor dorsalis muscle. This functional modification suggests that crushing hard-shelled marine bivalves requires a 'vice-like' compression bite in contrast to the shearing forces that are applied to weaker-shelled fiddler crabs by red drum and to freshwater snails by redear sunfish. PMID- 11003828 TI - Ventilatory and metabolic responses to hypoxia and sulphide in the lugworm Arenicola marina (L.). AB - We examined the effects of hypoxia and sulphide levels on the ventilatory activity of Arenicola marina and determined whether ventilation compensates for oxygen deficiency and affects the mode of energy provision. A. marina ventilated intermittently, irrespective of ambient P(O2) and sulphide concentration. The ventilation rate was 28.5+/-16 ml h(-1) g(-1) wet mass during normoxia, but increased to 175+/-60% of this value during moderate hypoxia, during which aerobic energy metabolism was maintained. Below a P(O2) of 6.2 kPa, A. marina reduced the ventilated volume to 54+/-16% of the normoxic value and became anaerobic, as indicated by the accumulation of succinate and strombine. Incubation with 27 micromol l(-1) ambient sulphide had no effect on the normoxic and hypoxic ventilation rates or on the P(O2) below which anaerobiosis started (P(cM)). Increased sulphide concentrations reduced the ventilation rate and shifted the P(cM) towards a higher P(O2) below 10.7 kPa. Sulphide diffused into the body and was at least partially detoxified to thiosulphate when oxygen was present. Under normoxia, sulphide accumulated in the body wall tissue and coelomic fluid when ambient sulphide levels exceeded 117 micromol l(-1) and 216 micromol l(-1), respectively. A decrease in P(O2) in the presence of 27 or 117 micromol l(-1) ambient sulphide had no significant effect on sulphide accumulation. PMID- 11003829 TI - K(+) currents in cultured neurones from a polyclad flatworm. AB - Cells from the brain of the polyclad flatworm Notoplana atomata were dispersed and maintained in primary culture for up to 3 weeks. Whole-cell patch-clamp of presumed neurones revealed outwardly directed K(+) currents that comprised, in varying proportions, a rapidly activating (time constant tau =0.94+/-0.79 ms; N=15) and inactivating ( tau =26.1+/-1.9 ms; N=22) current and a second current that also activated rapidly ( tau =1.1+/-0.2 ms; N=9) (means +/- s.e.m.) but did not inactivate within 100 ms. Both current types activated over similar voltage ranges. Activation and steady-state inactivation overlap and are markedly rightward-shifted compared with most Shaker-like currents (half-activation of 16.9+/-1. 9 mV, N=7, half-inactivation of -35.4+/-3.0 mV, N=5). Recovery from inactivation was rapid (50+/-2.5 ms at -90 mV). Both currents were unaffected by tetraethylammonium (25 mmol l(-1)), whereas 4-aminopyridine (10 mmol l(-1)) selectively blocked the inactivating current. The rapidly inactivating current, like cloned K(+) channels from cnidarians and certain cloned K(+) channels from molluscs and the Kv3 family of vertebrate channels, differed from most A-type K(+) currents reported to date. These findings suggest that K(+) currents in Notoplana atomata play novel roles in shaping excitability properties. PMID- 11003830 TI - The mechanisms of urea transport by early life stages of rainbow trout (Oncorhynchus mykiss). AB - We tested the hypothesis that urea transport in rainbow trout (Oncorhynchus mykiss) embryos is dependent, in part, on a bidirectional urea-transport protein. Acute exposure to phloretin and urea analogs [acetamide, thiourea, 1,(4 nitrophenyl)-2-thiourea] reversibly inhibited urea excretion from the embryos to the external water. Unidirectional urea influx was inhibited by acetamide and thiourea, with IC(50) values of 0.04 and 0.05 mmol l(-1), respectively. Influx of urea from the external water to the embryo tended to saturate at elevated external urea concentrations (V(max)=10.50 nmol g(-1) h(-1); K(m)=2 mmol l(-1)). At very high urea concentrations (20 mmol l(-1)), however, a second, non saturable component was apparent. These results indicate that urea excretion in trout embryos is dependent, in part, on a phloretin-sensitive facilitated urea transporter similar to that reported in mammalian inner medullary collecting ducts and elasmobranch kidney. PMID- 11003831 TI - Hhex and scl function in parallel to regulate early endothelial and blood differentiation in zebrafish. AB - During embryogenesis, endothelial and blood precursors are hypothesized to arise from a common progenitor, the hemangioblast. Several genes that affect the differentiation of, or are expressed early in, both the endothelial and blood lineages may in fact function at the level of the hemangioblast. For example, the zebrafish cloche mutation disrupts the differentiation of both endothelial and blood cells. The transcription factor gene scl is expressed in both endothelial and blood lineages from an early stage and can regulate their differentiation. Here we report that in zebrafish the homeobox gene hhex (previously called hex) is also expressed in endothelial and blood lineages from an early stage. We find that hhex expression in these lineages is significantly reduced in cloche mutant embryos, indicating that hhex functions downstream of cloche to regulate endothelial and blood differentiation. Ectopic expression of hhex through injection of a DNA construct leads to the premature and ectopic expression of early endothelial and blood differentiation genes such as fli1, flk1 and gata1, indicating that Hhex can positively regulate endothelial and blood differentiation. However, analysis of a hhex deficiency allele shows that hhex is not essential for early endothelial and blood differentiation, suggesting that another gene, perhaps scl, compensates for the absence of Hhex function. Furthermore, we find that hhex and scl can induce each other's expression, suggesting that these two genes cross-regulate each other during early endothelial and blood differentiation. Together, these data provide the initial framework of a pathway that can be used to further integrate the molecular events regulating hemangioblast differentiation. PMID- 11003832 TI - Requirements of Lim1, a Drosophila LIM-homeobox gene, for normal leg and antennal development. AB - During Drosophila leg development, the distal-most compartment (pretarsus) and its immediate neighbour (tarsal segment 5) are specified by a pretarsus-specific homeobox gene, aristaless, and tarsal-segment-specific Bar homeobox genes, respectively; the pretarsus/tarsal-segment boundary is formed by antagonistic interactions between Bar and pretarsus-specific genes that include aristaless (Kojima, T., Sato, M. and Saigo, K. (2000) Development 127, 769-778). Here, we show that Drosophila Lim1, a homologue of vertebrate Lim1 encoding a LIM homeodomain protein, is involved in pretarsus specification and boundary formation through its activation of aristaless. Ectopic expression of Lim1 caused aristaless misexpression, while aristaless expression was significantly reduced in Lim1-null mutant clones. Pretarsus Lim1 expression was negatively regulated by Bar and abolished in leg discs lacking aristaless activity, which was associated with strong Bar misexpression in the presumptive pretarsus. No Lim1 misexpression occurred upon aristaless misexpression. The concerted function of Lim1 and aristaless was required to maintain Fasciclin 2 expression in border cells and form a smooth pretarsus/tarsal-segment boundary. Lim1 was also required for femur, coxa and antennal development. PMID- 11003833 TI - Spatial specification of mammalian eye territories by reciprocal transcriptional repression of Pax2 and Pax6. AB - We have studied the molecular basis of the Pax2 and Pax6 function in the establishment of visual system territories. Loss-of-function mutants have revealed crucial roles for Pax2 in the generation of the optic stalk and for Pax6 in the development of the optic cup. Ectopic expression of Pax6 in the optic stalk under control of Pax2 promoter elements resulted in a shift of the optic cup/optic stalk boundary indicated by the presence of retinal pigmented cells on the optic stalk. By studying mouse embryos at early developmental stages we detected an expansion of Pax2 expression domain in the Pax6(-/-) mutant and of Pax6 expression domain in the Pax2(-/-) embryo. These results suggest that the position of the optic cup/optic stalk boundary depends on Pax2 and Pax6 expression, hinting at a possible molecular interaction. Using gel shift experiments, we confirmed the presence of Pax2- and Pax6-binding sites on the retina enhancer of the Pax6 gene and on the Pax2 upstream control region, respectively. Co-transfection experiments revealed a reciprocal inhibition of Pax2 promoter/enhancer activity by Pax6 protein and vice versa. Based on our findings, we propose a model for Pax gene regulation that establishes the proper spatial regionalization of the mammalian visual system. PMID- 11003834 TI - Positive and negative interactions of GDNF, NTN and ART in developing sensory neuron subpopulations, and their collaboration with neurotrophins. AB - Glial cell line-derived neurotrophic factor (GDNF), neurturin (NTN) and neublastin/artemin (ART) are distant members of the transforming growth factor beta family, and have been shown to elicit neurotrophic effects upon several classes of peripheral and central neurons. Limited information from in vitro and expression studies has also substantiated a role for GDNF family ligands in mammalian somatosensory neuron development. Here, we show that although dorsal root ganglion (DRG) sensory neurons express GDNF family receptors embryonically, they do not survive in response to their ligands. The regulation of survival emerges postnatally for all GDNF family ligands. GDNF and NTN support distinct subpopulations that can be separated with respect to their expression of GDNF family receptors, whereas ART supports neurons in populations that are also responsive to GDNF or NTN. Sensory neurons that coexpress GDNF family receptors are medium sized, whereas small-caliber nociceptive cells preferentially express a single receptor. In contrast to brain-derived neurotrophic factor (BDNF) dependent neurons, embryonic nerve growth factor (NGF)-dependent nociceptive neurons switch dependency to GDNF, NTN and ART postnatally. Neurons that survive in the presence of neurotrophin 3 (NT3) or neurotrophin 4 (NT4), including proprioceptive afferents, Merkel end organs and D-hair afferents, are also supported by GDNF family ligands neonatally, although at postnatal stages they lose their dependency on GDNF and NTN. At late postnatal stages, ART prevents survival elicited by GDNF and NTN. These data provide new insights on the roles of GDNF family ligands in sensory neuron development. PMID- 11003835 TI - A role for GATA5 in Xenopus endoderm specification. AB - The endoderm gives rise to the gut and tissues that develop as outgrowths of the gut tube, including the lungs, liver and pancreas. Here we show that GATA5, a zinc-finger transcription factor, is expressed in the yolk-rich vegetal cells of Xenopus embryos from the early gastrula stage onwards, when these cells become committed to form endoderm. At mid-gastrula stages, GATA5 is restricted to the sub-blastoporal endoderm and is the first molecular marker for this subset of endodermal cells so far identified. We show that GATA4 and GATA5 are potent inducers of endodermal marker genes in animal cap assays, while other GATA factors induce these genes only weakly, if at all. When injected into the dorsal marginal zone, GATA5 respecifies prospective mesoderm towards an endodermal fate, thereby disrupting the convergence and extension movements normally undergone by the dorsal mesoderm. The resulting phenotype is very similar to those seen after injection of dominant negative versions of the FGF-receptor or the T-box transcription factor, Xbra and can be rescued by eFGF. The ability of GATA5 to respecify ectodermal and mesodermal cells towards endoderm suggests an important role for GATA5 in the formation of this germlayer. In animal cap assays, GATA5 is induced by concentrations of activin above those known to induce dorsal mesoderm and heart, in an FGF-independent manner. These data indicate that the emerging view for endodermal induction in general, namely that it is specified by high levels of TGF-beta in the absence of FGF signalling, is specifically true for sub blastoporal endoderm. PMID- 11003836 TI - Genetic control of dorsal-ventral identity in the telencephalon: opposing roles for Pax6 and Gsh2. AB - We have examined the genetic mechanisms that regulate dorsal-ventral identity in the embryonic mouse telencephalon and, in particular, the specification of progenitors in the cerebral cortex and striatum. The respective roles of Pax6 and Gsh2 in cortical and striatal development were studied in single and double loss of-function mouse mutants. Gsh2 gene function was found to be essential to maintain the molecular identity of early striatal progenitors and in its absence the ventral telencephalic regulatory genes Mash1 and Dlx are lost from most of the striatal germinal zone. In their place, the dorsal regulators, Pax6, neurogenin 1 and neurogenin 2 are found ectopically. Conversely, Pax6 is required to maintain the correct molecular identity of cortical progenitors. In its absence, neurogenins are lost from the cortical germinal zone and Gsh2, Mash1 and Dlx genes are found ectopically. These reciprocal alterations in cortical and striatal progenitor specification lead to the abnormal development of the cortex and striatum observed in Pax6 (small eye) and Gsh2 mutants, respectively. In support of this, double homozygous mutants for Pax6 and Gsh2 exhibit significant improvements in both cortical and striatal development compared with their respective single mutants. Taken together, these results demonstrate that Pax6 and Gsh2 govern cortical and striatal development by regulating genetically opposing programs that control the expression of each other as well as the regionally expressed developmental regulators Mash1, the neurogenins and Dlx genes in telencephalic progenitors. PMID- 11003837 TI - Correlation of diversity of leg morphology in Gryllus bimaculatus (cricket) with divergence in dpp expression pattern during leg development. AB - Insects can be grouped into mainly two categories, holometabolous and hemimetabolous, according to the extent of their morphological change during metamorphosis. The three thoracic legs, for example, are known to develop through two overtly different pathways: holometabolous insects make legs through their imaginal discs, while hemimetabolous legs develop from their leg buds. Thus, how the molecular mechanisms of leg development differ from each other is an intriguing question. In the holometabolous long-germ insect, these mechanisms have been extensively studied using Drosophila melanogaster. However, little is known about the mechanism in the hemimetabolous insect. Thus, we studied leg development of the hemimetabolous short-germ insect, Gryllus bimaculatus (cricket), focusing on expression patterns of the three key signaling molecules, hedgehog (hh), wingless (wg) and decapentaplegic (dpp), which are essential during leg development in Drosophila. In Gryllus embryos, expression of hh is restricted in the posterior half of each leg bud, while dpp and wg are expressed in the dorsal and ventral sides of its anteroposterior (A/P) boundary, respectively. Their expression patterns are essentially comparable with those of the three genes in Drosophila leg imaginal discs, suggesting the existence of the common mechanism for leg pattern formation. However, we found that expression pattern of dpp was significantly divergent among Gryllus, Schistocerca (grasshopper) and Drosophila embryos, while expression patterns of hh and wg are conserved. Furthermore, the divergence was found between the pro/mesothoracic and metathoracic Gryllus leg buds. These observations imply that the divergence in the dpp expression pattern may correlate with diversity of leg morphology. PMID- 11003838 TI - Developmental regulation of GDNF response and receptor expression in the enteric nervous system. AB - The development of the enteric nervous system is dependent upon the actions of glial cell line-derived neurotrophic factor (GDNF) on neural crest-derived precursor cells in the embryonic gut. GDNF treatment of cultured enteric precursor cells leads to an increase in the number of neurons that develop and/or survive. Here we demonstrate that, although GDNF promoted an increase in neuron number at all embryonic ages examined, there was a developmental shift from a mitogenic to a trophic response by the developing enteric neurons. The timing of this shift corresponded to developmental changes in gut expression of GFR alpha 1, a co-receptor in the GDNF-Ret signaling complex. GFR alpha-1 was broadly expressed in the gut at early developmental stages, at which times soluble GFR alpha-1 was released into the medium by cultured gut cells. At later times, GFR alpha-1 became restricted to neural crest-derived cells. GFR alpha-1 could participate in GDNF signaling when expressed in cis on the surface of enteric precursor cells, or as a soluble protein. The GDNF-mediated response was greater when cell surface, compared with soluble, GFR alpha-1 was present, with the maximal response seen the presence of both cis and trans forms of GFR alpha-1. In addition to contributing to GDNF signaling, cell-surface GFR alpha-1 modulated the specificity of interactions between GDNF and soluble GFR alphas. These experiments demonstrate that complex, developmentally regulated, signaling interactions contribute to the GDNF-dependent development of enteric neurons. PMID- 11003839 TI - Gli2 functions in FGF signaling during antero-posterior patterning. AB - Patterning along the anteroposterior (A-P) axis involves the interplay of secreted and transcription factors that specify cell fates in the mesoderm and neuroectoderm. While FGF and homeodomain proteins have been shown to play different roles in posterior specification, the network coordinating their effects remains elusive. Here we have analyzed the function of Gli zinc-finger proteins in mesodermal A-P patterning. We find that Gli2 is sufficient to induce ventroposterior development, functioning in the FGF-brachyury regulatory loop. Gli2 directly induces brachyury, a gene required and sufficient for mesodermal development, and Gli2 is in turn induced by FGF signaling. Moreover, the homeobox gene Xhox3, a critical determinant of posterior development, is also directly regulated by Gli2. Gli3, but not Gli1, has an activity similar to that of Gli2 and is expressed in ventroposterior mesoderm after Gli2. These findings uncover a novel function of Gli proteins, previously only known to mediate hedgehog signals, in the maintenance and patterning of the embryonic mesoderm. More generally, our results suggest a molecular basis for an integration of FGF and hedgehog inputs in Gli-expressing cells that respond to these signals. PMID- 11003840 TI - Wingless and its signalling pathway have common and separable functions during tracheal development. AB - The Drosophila tracheal tree consists of a tubular network of epithelial branches that constitutes the respiratory system. Groups of tracheal cells migrate towards stereotyped directions while they acquire specific tracheal fates. This work shows that the wingless/WNT signalling pathway is needed within the tracheal cells for the formation of the dorsal trunk and for fusion of the branches. These functions are achieved through the regulation of target genes, such as spalt in the dorsal trunk and escargot in the fusion cells. The pathway also aids tracheal invagination and helps guide the ganglionic branch. Moreover the wingless/WNT pathway displays antagonistic interactions with the DPP (decapentaplegic) pathway, which regulates branching along the dorsoventral axis. Remarkably, the wingless gene itself, acting through its canonical pathway, seems not to be absolutely required for all these tracheal functions. However, the artificial overexpression of wingless in tracheal cells mimics the overexpression of a constitutively activated armadillo protein. The results suggest that another gene product, possibly a WNT, could help to trigger the wingless cascade in the developing tracheae. PMID- 11003842 TI - Control of tracheal tubulogenesis by Wingless signaling. AB - The tubular epithelium of the Drosophila tracheal system forms a network with a stereotyped pattern consisting of cells and branches with distinct identity. The tracheal primordium undergoes primary branching induced by the FGF homolog Branchless, differentiates cells with specialized functions such as fusion cells, which perform target recognition and adhesion during branch fusion, and extends branches toward specific targets. Specification of a unique identity for each primary branch is essential for directed migration, as a defect in either the EGFR or the Dpp pathway leads to a loss of branch identity and the misguidance of tracheal cell migration. Here, we investigate the role of Wingless signaling in the specification of cell and branch identity in the tracheal system. Wingless and its intracellular signal transducer, Armadillo, have multiple functions, including specifying the dorsal trunk through activation of Spalt expression and inducing differentiation of fusion cells in all fusion branches. Moreover, we show that Wingless signaling regulates Notch signaling by stimulating delta expression at the tip of primary branches. These activities of Wingless signaling together specify the shape of the dorsal trunk and other fusion branches. PMID- 11003841 TI - MES-1, a protein required for unequal divisions of the germline in early C. elegans embryos, resembles receptor tyrosine kinases and is localized to the boundary between the germline and gut cells. AB - During Caenorhabditis elegans embryogenesis the primordial germ cell, P(4), is generated via a series of unequal divisions. These divisions produce germline blastomeres (P(1), P(2), P(3), P(4)) that differ from their somatic sisters in their size, fate and cytoplasmic content (e.g. germ granules). mes-1 mutant embryos display the striking phenotype of transformation of P(4) into a muscle precursor, like its somatic sister. A loss of polarity in P(2) and P(3) cell specific events underlies the Mes-1 phenotype. In mes-1 embryos, P(2) and P(3) undergo symmetric divisions and partition germ granules to both daughters. This paper shows that mes-1 encodes a receptor tyrosine kinase-like protein, though it lacks several residues conserved in all kinases and therefore is predicted not to have kinase activity. Immunolocalization analysis shows that MES-1 is present in four- to 24-cell embryos, where it is localized in a crescent at the junction between the germline cell and its neighboring gut cell. This is the region of P(2) and P(3) to which the spindle and P granules must move to ensure normal division asymmetry and cytoplasmic partitioning. Indeed, during early stages of mitosis in P(2) and P(3), one centrosome is positioned adjacent to the MES-1 crescent. Staining of isolated blastomeres demonstrated that MES-1 was present in the membrane of the germline blastomeres, consistent with a cell-autonomous function. Analysis of MES-1 distribution in various cell-fate and patterning mutants suggests that its localization is not dependent on the correct fate of either the germline or the gut blastomere but is dependent upon correct spatial organization of the embryo. Our results suggest that MES-1 directly positions the developing mitotic spindle and its associated P granules within P(2) and P(3), or provides an orientation signal for P(2)- and P(3)-specific events. PMID- 11003843 TI - The SPIRAL genes are required for directional control of cell elongation in Aarabidopsis thaliana. AB - Cells at the elongation zone expand longitudinally to form the straight central axis of plant stems, hypocotyls and roots, and transverse cortical microtubule arrays are generally recognized to be important for the anisotropic growth. Recessive mutations in either of two Arabidopsis thaliana SPIRAL loci, SPR1 or SPR2, reduce anisotropic growth of endodermal and cortical cells in roots and etiolated hypocotyls, and induce right-handed helical growth in epidermal cell files of these organs. spr2 mutants additionally show right-handed twisting in petioles and petals. The spr1spr2 double mutant's phenotype is synergistic, suggesting that SPR1 and SPR2 act on a similar process but in separate pathways in controlling cell elongation. Interestingly, addition of a low dose of either of the microtubule-interacting drugs propyzamide or taxol in the agar medium was found to reduce anisotropic expansion of endodermal and cortical cells at the root elongation zone of wild-type seedlings, resulting in left-handed helical growth. In both spiral mutants, exogenous application of these drugs reverted the direction of the epidermal helix, in a dose-dependent manner, from right-handed to left-handed; propyzamide at 1 microM and taxol at 0.2-0.3 microM effectively suppressed the cell elongation defects of spiral seedlings. The spr1 phenotype is more pronounced at low temperatures and is nearly suppressed at high temperatures. Cortical microtubules in elongating epidermal cells of spr1 roots were arranged in left-handed helical arrays, whereas the highly isotropic cortical cells of etiolated spr1 hypocotyls showed microtubule arrays with irregular orientations. We propose that a microtubule-dependent process and SPR1/SPR2 act antagonistically to control directional cell elongation by preventing elongating cells from potential twisting. Our model may have implicit bearing on the circumnutation mechanism. PMID- 11003844 TI - Modular long-range regulation of Myf5 reveals unexpected heterogeneity between skeletal muscles in the mouse embryo. AB - The myogenic factor Myf5 plays a key role in muscle cell determination, in response to signalling cascades that lead to the specification of muscle progenitor cells. We have adopted a YAC transgenic approach to identify regulatory sequences that direct the complex spatiotemporal expression of this gene during myogenesis in the mouse embryo. Important regulatory regions with distinct properties are distributed over 96 kb upstream of the Myf5 gene. The proximal 23 kb region directs early expression in the branchial arches, epaxial dermomyotome and in a central part of the myotome, the epaxial intercalated domain. Robust expression at most sites in the embryo where skeletal muscle forms depends on an enhancer-like sequence located between -58 and -48 kb from the Myf5 gene. This element is active in the epaxial and hypaxial myotome, in limb muscles, in the hypoglossal chord and also at the sites of Myf5 transcription in prosomeres p1 and p4 of the brain. However later expression of Myf5 depends on a more distal region between -96 and -63 kb, which does not behave as an enhancer. This element is necessary for expression in head muscles but strikingly only plays a role in a subset of trunk muscles, notably the hypaxially derived ventral body muscles and also those of the diaphragm and tongue. Transgene expression in limb muscle masses is not affected by removal of the -96/-63 region. Epaxially derived muscles and some hypaxial muscles, such as the intercostals and those of the limb girdles, are also unaffected. This region therefore reveals unexpected heterogeneity between muscle masses, which may be related to different facets of myogenesis at these sites. Such regulatory heterogeneity may underlie the observed restriction of myopathies to particular muscle subgroups. PMID- 11003845 TI - Mab-3 is a direct tra-1 target gene regulating diverse aspects of C. elegans male sexual development and behavior. AB - Sex determination is controlled by global regulatory genes, such as tra-1 in Caenorhabditis elegans, Sex lethal in Drosophila, or Sry in mammals. How these genes coordinate sexual differentiation throughout the body is a key unanswered question. tra-1 encodes a zinc finger transcription factor, TRA-1A, that regulates, directly or indirectly, all genes required for sexual development. mab 3 (male abnormal 3), acts downstream of tra-1 and is known to be required for sexual differentiation of at least two tissues. mab-3 directly regulates yolk protein transcription in the intestine and specifies male sense organ differentiation in the nervous system. It encodes a transcription factor related to the products of the Drosophila sexual regulator doublesex (dsx), which also regulates yolk protein transcription and male sense-organ differentiation. The similarities between mab-3 and dsx led us to suggest that some aspects of sex determination may be evolutionarily conserved. Here we find that mab-3 is also required for expression of male-specific genes in sensory neurons of the head and tail and for male interaction with hermaphrodites. These roles in male development and behavior suggest further functional similarity to dsx. In male sensory ray differentiation we find that MAB-3 acts synergistically with LIN-32, a neurogenic bHLH transcription factor. Expression of LIN-32 is spatially restricted by the combined action of the Hox gene mab-5 and the hairy homolog lin 22, while MAB-3 is expressed throughout the lateral hypodermis. Finally, we find that mab-3 transcription is directly regulated in the intestine by TRA-1A, providing a molecular link between the global regulatory pathway and terminal sexual differentiation. PMID- 11003846 TI - Lactational competence and involution of the mouse mammary gland require plasminogen. AB - Urokinase-type plasminogen activator expression is induced in the mouse mammary gland during development and post-lactational involution. We now show that primiparous plasminogen-deficient (Plg(-/-)) mice have seriously compromised mammary gland development and involution. All mammary glands were underdeveloped and one-quarter of the mice failed to lactate. Although the glands from lactating Plg(-/-) mice were initially smaller, they failed to involute after weaning, and in most cases they failed to support a second litter. Alveolar regression was markedly reduced and a fibrotic stroma accumulated in Plg(-/-) mice. Nevertheless, urokinase and matrix metalloproteinases (MMPs) were upregulated normally in involuting glands of Plg(-/-) mice, and fibrin did not accumulate in the glands. Heterozygous Plg(+/-) mice exhibited haploinsufficiency, with a definite, but less severe mammary phenotype. These data demonstrate a critical, dose-dependent requirement for Plg in lactational differentiation and mammary gland remodeling during involution. PMID- 11003847 TI - Developmental role of the SNF1-related kinase Hunk in pregnancy-induced changes in the mammary gland. AB - The steroid hormones 17 beta-estradiol and progesterone play a central role in the pathogenesis of breast cancer and regulate key phases of mammary gland development. This suggests that developmental regulatory molecules whose activity is influenced by ovarian hormones may also contribute to mammary carcinogenesis. In a screen designed to identify protein kinases expressed in the mammary gland, we previously identified a novel SNF1-related serine/threonine kinase, Hunk (hormonally upregulated Neu-associated kinase). During postnatal mammary development, Hunk mRNA expression is restricted to a subset of mammary epithelial cells and is temporally regulated with highest levels of expression occurring during early pregnancy. In addition, treatment of mice with 17 beta-estradiol and progesterone results in the rapid and synergistic upregulation of Hunk expression in a subset of mammary epithelial cells, suggesting that the expression of this kinase may be regulated by ovarian hormones. Consistent with the tightly regulated pattern of Hunk expression during pregnancy, mammary glands from transgenic mice engineered to misexpress Hunk in the mammary epithelium manifest temporally distinct defects in epithelial proliferation and differentiation during pregnancy, and fail to undergo normal lobuloalveolar development. Together, these observations suggest that Hunk may contribute to changes in the mammary gland that occur during pregnancy in response to ovarian hormones. PMID- 11003848 TI - Attractive and repulsive interactions between female and male gametophytes in Arabidopsis pollen tube guidance. AB - Sexual reproduction in plants, unlike that of animals, requires the action of multicellular haploid gametophytes. The male gametophyte (pollen tube) is guided to a female gametophyte through diploid sporophytic cells in the pistil. While interactions between the pollen tube and diploid cells have been described, little is known about the intercellular recognition systems between the pollen tube and the female gametophyte. In particular, the mechanisms that enable only one pollen tube to interact with each female gametophyte, thereby preventing polysperm, are not understood. We isolated female gametophyte mutants named magatama (maa) from Arabidopsis thaliana by screening for siliques containing half the normal number of mature seeds. In maa1 and maa3 mutants, in which the development of the female gametophyte was delayed, pollen tube guidance was affected. Pollen tubes were directed to mutant female gametophytes, but they lost their way just before entering the micropyle and elongated in random directions. Moreover, the mutant female gametophytes attracted two pollen tubes at a high frequency. To explain the interaction between gametophytes, we propose a monogamy model in which a female gametophyte emits two attractants and prevents polyspermy. This prevention process by the female gametophyte could increase a plant's inclusive fitness by facilitating the fertilization of sibling female gametophytes. In addition, repulsion between pollen tubes might help prevent polyspermy. The reproductive isolations observed in interspecific crosses in Brassicaceae are also consistent with the monogamy model. PMID- 11003849 TI - [Gratitude to the past and trust in the future] [In Process Citation] PMID- 11003850 TI - [Omega and alpha, preserving and thriving] [In Process Citation] PMID- 11003851 TI - [Treatment of benign vascular lesions with 585 nm pulsed dye laser]. PMID- 11003852 TI - [Laser treatment of cutaneous vascular lesions in childhood: prospective analysis of our experience with 95 children]. AB - BACKGROUND: Recently the flashlamp-pumped dye laser has improved treatment of cutaneous vascular lesions results since this laser acts much more vessel specific than other laser (argon, Nd:Yag). The objective of this manuscript is to analyze our experience in the treatment of vascular lesions in infancy with the pulsed dye laser. MATERIAL AND METHODS: Ninety-five patients 18 years with vascular lesions (85 nevus flammeus and 10 hemangiomas) were treated prospectively with the flashlamp-pumped dye laser. The patients received therapy until the lesion was almost clear, or until the lesion failed to respond. All patients were examined during the course of their treatment to assess the clinical evolution and the incidence of adverse effects associated with the use of the laser. RESULTS: The study consisted of 54 females and 41 males ranged in age from 3 months to 18 years, with the average age being 7.3+/-5.2 years. Only six children were treated with less than six months of age. Seventy-eight vascular lesions were located on the face, nine on the neck, eight on the extremities, five on the trunk, and two on other regions (perineal area and scalp). The number of treatment sessions needed for the resolution of the cutaneous vascular lesion (in 52 nevus flammeus and 10 hemangiomas, at the moment) was 9.8+/-7.5 (range 1 to 36), with variations in relation to the type of vascular lesion (more sessions in nevus flammeus than hemangiomas) and their anatomical location (more sessions in the face and less in shoulder, arms and lips). The adverse effects were slights and transitories. CONCLUSIONS: Laser therapy is useful and safe in the treatment of cutaneous vascular lesions in infancy. Major determinants of treatment response are type of cutaneous vascular lesion and location, and also size and patient's age. PMID- 11003853 TI - [Clinical and radiological characteristics of primary pulmonary tuberculosis in adolescents]. AB - OBJECTIVE: The aim of this study was to investigate the incidence, clinical and radiological particularities of primary pulmonary tuberculosis in children between 10 to 16 year-old in our community. PATIENTS AND METHODS: The medical records and chest X-ray of all children under 16 years old diagnosed of primary pulmonary tuberculosis between 1982 to 1996, and a similar characteristics control group diagnosed of bacterial pneumonia were reviewed. The difference between teenager with tuberculosis and pneumonia and between patients with tuberculosis under 10 years and order were analysed. RESULTS: Between January 1982 and December 1996, 83 children with age range from 10 to 16 years were diagnosed of primary pulmonary tuberculosis. The incidence has been 15,5 cases/100000 h < 16 years/year. The primary pulmonary tuberculosis whole incidence in children under 16 year-old has been 17 cases/100000 h/year (182 cases). The adolescents fits to 40.5% of the hold group. 51.2% were girls and 49.8% were boys. Middle age has been 14.2+/-0.18 years with a progressive increased with the age. Anorexia (38% vs. 16.6%) and asthenia (38% vs 19.4%) have been more frequents in teenagers with tuberculosis. Cough has been the most frequent symptom in both groups (61.9% and 77.7%), and respiratory distress has been the most characteristic of tuberculosis (17.8% vs 2.7%). Thoracic pain was more frequent in patients with tuberculosis 10 years or older (57.8% and 7.8%). Laboratory values are not specific. Pleural effusion (46.4% vs 22.2%) and mediastinal lymphadenophaty (60.1% vs. 40.4%) were the most characteristic X-ray finding in teenagers and children under 10 years with primary pulmonary tuberculosis respectively. CONCLUSIONS: Primary pulmonary tuberculosis has a high incidence in the teenager population in our community with a progressive increased with the age. The clinical and laboratory values are unspecific in this age group. The most characteristic X-ray finding is pleural effusion (46.4% of the patients). PMID- 11003854 TI - [Treatment of suprastomal tracheomalacia by anterior cricoid suspension]. AB - BACKGROUND: One of the reasons of failure to decannulate patients after airway surgery or long-term tracheostomy is suprastomal tracheomalacia. OBJECTIVES: Evaluate the results obtained in the treatment of suprastomal tracheomalacia by anterior cricoid suspension. MATERIAL AND METHODS: We present 8 patients with suprastomal tracheomalacia associated to long-term tracheostomy corrected by this technique. Prior to surgery, other causes of airway obstruction, as well as the presence of gastroesophageal reflux disease should be ruled out. Results 5 females and 3 males wer treated by this technique. Mean age at surgery was 4,1 +/ 0,97 years (range, 1,5-8,8 years). anteroposterior suprastomal collapse occluding 75% of the trachea was observed in 5 cases, and 75-90% in other three. All the patients were extubated successfully, although in one case stridor recurred because of residual suprastomal malacia that required a new surgical procedure. After a follow-up of 38,7+/-7,39 months (range, 1 month-4,6 years), the patients are decannulated and going well. Conclusions It is an safe surgical technique, easily reproducible and with low costs. PMID- 11003855 TI - [Technetium-99m-dimercaptosuccinic acid (DMSA) scintigraphy in the first febrile urinary tract infection in children]. AB - OBJECTIVE: The objective of this study was to evaluate the findings of 99mTc-DMSA renal scintigraphy in children with their first acute febrile urinary tract infection in relation with several clinical-biological parameters and other imaging studies and a long-term follow up. MATERIAL AND METHODS: 103 children between 1 month and 10 years old with their first acute febrile urinary tract infection were studied by means of clinical and laboratory assessment, renal ultrasonography and 99mTc DMSA renal scintigraphy in the acute illness period. Patients were divided into two groups, those under 2 years old (n = 63) and those over 2 years old (n = 40). RESULTS: Cortical scintigraphy showed renal changes in 56 patients (54,5%). Children over 2 years of age had a higher incidence of renal lesions than younger children (39,7% vs. 77,5%). Ultrasonography measuring renal volume showed a simple concordance with scintigraphy in 64% of the patients. Reflux was demonstrated in 23 patients (22,3%) without differences between patients with normal or abnormal cortical scintigraphy. After having studied the factors that were associated independently to the pathological results in the scintigraphy through a logistical multivariant regression, it was observed that the child's age, the elevation of the PCR and the alterations of the renal volume were significant according to statistics and also associated to a larger amount of pathological DMSA scintigraphy. CONCLUSIONS: DMSA scintigraphy associated with other diagnostic methods can improve the sensitivity and specificity to establish renal damage stage. During the follow-up the acute lesions disappeared or improved in all cases and the chronic lesions have not been modificated. PMID- 11003856 TI - [Suicide in pediatric patients: 30 cases]. AB - OBJECTIVE: The aim of this study was to evaluate suicide attempts in pediatric population and analyze the morbimortality. PATIENTS AND METHODS: We have reviewed thirty reports from twenty six patients hospitalized in UCIP by suicide attempts with an average age of 15.5 years (range 11-19) between January 1991 and April 1999. Results The following variables were analyzed: age, sex, precipitating suicidal event, number of suicide attempts, symptoms, method, treatment and mortality. The age group with more incidence was between 14 and 17 years with 73% (22 cases). The majority were girls (96%). There were prior suicide attempt in 40% (12 cases). Psychiatric disorder more frequently associated was anorexia nervosa in 70%. The most common method was self-poisoning (93%). Neurologic symptoms were seen in 20 cases. The more used treatment was activated coal. The clinical outcome was favourable and only two patients died. Conclusions The suicide attempts are happening more frequently in the adolescence in the last years. Pediatricians have to be prepared to recognize and provide prompt attention to acute medical, familiar and social problems. PMID- 11003857 TI - [Cardiac rhabdomyomas in tuberous sclerosis: clinical symptoms and course in 18 cases diagnosed in childhood]. AB - OBJECTIVE: Tuberous sclerosis is a neurocutaneous syndrome with autosomal dominant inheritance. The reported incidence is 1:10000. Recent echocardiographic studies showed a 50% to 64% incidence of cardiac rhabdomyomas in patients with tuberous sclerosis. The objective is to assess the history of cardiac rhabdomyomas in infants and children with tuberous sclerosis. METHODS: From 1970 to 1998, 39 patients were diagnosticated of tuberous sclerosis. Cardiac rhabdomyomas were present in 18 of them, 9 girls and 9 boys. Ecocardiografic follow up ranged from 1 month to 14 years. RESULTS: Echocardiographic studies showed rhabdomyomas located in the interventricular septum, ventricles, right and left atrium. A newborn, had subaortic obstruction related to a tumor who precised surgery excision. Standard electrocardiogram showed different disturbances: premature auricular and ventricular contractions, conduccion disturbances and pre excitation syndromes. Two patients, both neonates, had severe arrhythmias, one of them debuted with supraventricular tachycardia due to an accessory hidden left atrio-ventricular pathway who undergone successful radiofrequency ablation; and the other patient was diagnosticated prenatally of fetal complete heart block and precised permanent pacemaker at birth. After a follow up of 5.1+/-4.5 years, spontaneous regression of the tumors was shown in 12 of the 18 patients with complete resolution in two of them. All of them are asymptomatic nowadays. CONCLUSIONS: Our findings confirms that cardiac rhabdomyomas are most often a benign condition in which spontaneous regression is the rule and surgery is only recommended for patients with life threatening obstruction or refractory disrhythmias in the neonatal period when the tumor size is maximum. PMID- 11003858 TI - [Evaluation of a home care program for children with cancer]. AB - The home care team dependent from the pediatric oncology unit in our institution started working in April, 1997. We evaluate in this paper the medical activities accomplished in seventeen month experience. The team is constituted by a pediatric oncologist, two pediatric nurses and a clinical assistant with experience in the specialty. The geographic area we cover is la Communidad Valenciana. We directly attend children living in Valencia city and its metropolitan area. For the rest of patients, we coordinate the interventions of the local primary care teams and local hospitals. 127 patients have been admitted in the home care unit in 433 occasions. The immediate reasons for the admission were: early discharge from the hospital (61%), followed by the administration of antibiotics (18%) and chemotherapy (12%) at home. We attended 17 children in the terminal phase of their diseases. Five of them required opioid treatment for pain control. Six out of eight patients living in the area of direct intervention of the home care team died at home. The most common cause of discharge (73%) was the achievement of the goals planned when the patient was included in the program. Only in two cases (0.5%) we did not found enough cooperation from the parents and the treatment was completed in the hospital. This program has been well accepted by our patients and their parents and permits to shorten the stay in the hospital. PMID- 11003859 TI - [Multiple type I pseudohypoaldosteronism: neonatal management and outcome]. AB - Multiple type I pseudohypoaldosteronism (PHA-I) is an autosomal recessive condition with multiple target-organ unresponsiveness to aldosterone, manifested early after birth with severe salt-wasting and hyperkalemia. Case 1. Female infant born at term after an uneventful pregnancy. One female sibling died in the first week of life with hyperkalemia. The diagnosis of multiple PHA-I resulted from a picture of dehydratation, hyperkalemia and hyponatremia with increased plasma renin activity (PRA), plasma aldosterone and sweat electrolytes. The treatment consisted of salt and sodium bicarbonate supplements, restricted potassium intake, cation exchange resins and high fluid intake. During first year she was hospitalized for severe salt-losing crises. At 7 years of age, she needs salt and sodium bicarbonate supplements and cation exchange resins. She has a normal growth and neurodevelopment. Case 2. Seven-day female newborn with consanguinity in maternal family. Pregnancy and delivery were uncomplicated. On admission she was severely dehydrated with hyponatremia, hyperkalemia, metabolic acidosis and elevated PRA, plasma aldosterone and sweat electrolytes. She remained hospitalized for six months and she was dependent on high amounts of salt and sodium bicarbonate supplements, fluid intake and cation exchange resins. Growth and neurodevelopment are normal. CONCLUSIONS: Multiple PHA-I may be suspected in a newborn with salt-loss and hyperkalemia without glucocorticoid defect. The frequent episodes of dehydratation during the first year of life require long hospitalization. The improvement with age make possible an ambulatory control after the first year of life. PMID- 11003860 TI - [Neonatal cerebral thrombosis and deficit of factor V leiden]. AB - Background Cerebral venous thrombosis is an inusual disease in neonatal age. Increasing reports of this disorder had described since magnetic resonance angiography is used. Case report Newborn of apropriate seze for gestational age was delivered at 35 weeks of gestation. Refered a severe hipoxic-isquemic disease with multisistemic afectation. The second day of life presented disseminated intravascular coagulation with pulmonary bleeding. The third day, the infant developed seizures that required treatment with diazepam in continuous perfussion. MR angiography visualized superior sagital and transvers sinus thrombosis. Coagulation study detected factor V Leiden. Comments Frecuently venous cerebral thrombosis is presenting with lethargy and seizures. The most common vessels involved are sagital and transvers sinus. It is described in association with exogenous risk factors that increasing blood hyperviscosity and additional inhered coagulation dissorders such as defects on antihrombina III, protein C and S and activate protein C resistance. The last defect has a hight prevalence in subjects with trombosis events. PMID- 11003861 TI - [Adult respiratory distress in hemolytic-uremic syndrome secondary to pneumococcal meningitis]. PMID- 11003862 TI - [Periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis: report of three cases]. PMID- 11003863 TI - [Idiopathic arterial calcification in children: post-mortem diagnostic of a premature newborn]. PMID- 11003864 TI - [Acute parotitis caused by Epstein-Barr virus]. PMID- 11003865 TI - [Radiographic changes in cortical bone]. PMID- 11003866 TI - [Successful breast-feeding]. PMID- 11003867 TI - [Myocarditis caused by respiratory syncytial virus]. PMID- 11003868 TI - [Immunoglobulins to prevent varicella infection]. PMID- 11003869 TI - [Phytophotodermatitis]. PMID- 11003870 TI - [Rural gypsy population and breast-feeding. Is it an atypical population?]. PMID- 11003871 TI - [Streptococcus pyogenes invasive infection]. PMID- 11003873 TI - [II spanish congress on pediatric reumatology] [In Process Citation] PMID- 11003872 TI - [Eating disorders and their etiology. Not only fashion and media]. PMID- 11003874 TI - [Information] [In Process Citation] PMID- 11003875 TI - [Is smoking a pediatric epidemic?]. PMID- 11003876 TI - [Hormonal reference values for adrenocortical function in healthy children from Zaragoza]. AB - OBJECTIVE: Estimation of reference values for basal serum concentrations of adrenocorticotropic hormone (ACTH), cortisol, 11-deoxycortisol, 17-OH progesterone (17-OHP), plasma renin activity (PRA), aldosterone, -4 androstendione ( 4A) and dehydroepiandrosterone sulphate (DHA-S) in healthy children from Zaragoza. METHODS: Reference population were healthy children aged 0 to 14, with normal weight and height, living in the metropolitan area of Zaragoza (Spain). It is a transversal study. Reference values and ranges for ACTH, cortisol, 11-deoxycortisol, 17-OHP, PRA, aldosterone, 4A and DHA-S were estimated, and changes in concentrations were analyzed in relation to age, sex and puberal stage. RESULTS: Reference values have been classified by puberal stage and age in eleven groups for every sex: Tanner I (umbilical cordon, 3 days, 4-30 days, 1-6 months, 6 months-4 years, 4-7 years, 7-10 years, 10-14 years), Tanner II, Tanner III and Tanner IV-V. Sex did not influence ACTH, cortisol, 17 OHP and PRA concentrations, and there are punctual differences in 11 deoxycortisol, aldosterone, 4A and DHA-S levels. 17-OHP, 11-deoxycortisol and aldosterone concentrations significantly decreased from birth to 6 months-4 years and subsequently kept steady. The maximal concentration of ACTH, and ARP in blood cord significantly decreased until the period 6 months-4 years, and subsequent differences among different age groups, and between prepuberal and puberal groups are scarce. The highest concentration of 4A and DHA-S were observed in blood cord and third day of life, decreased until the lowest level in 6 months-4 years and progressively increased with age in prepuberty, and between prepuberty and puberty. The lowest concentration of cortisol was detected in 4-30 days, increased until 6 months-4 years and kept steady along the prepuberty and puberty. CONCLUSION: It is necessary that every population establish own reference values for ACTH, cortisol, 11-deoxycortisol, 17-OHP, PRA, aldosterone, 4A and DHA-S during infancy, childhood and adolescence, according to age, sex and puberal stage. PMID- 11003877 TI - [Secular trends of body adipose tissue in prepubertal children]. AB - OBJECTIVE: To determine whether there is a significant difference between anthropometric measurements of prepubertal children and the measurements of another sample with similar characteristics, studied fifteen years ago. MATERIAL AND METHODS: We studied 859 children, 453 males and 406 females, from the age of 5.0 to 10.0 years. Weight, height, arm circumference and skinfold thicknesses in the biceps, triceps, subscapular and suprailiac were determined. Body mass index, body density, total fat and percentage fat were calculated and their values compared with those obtained 15 years ago in 1,617 children with similar characteristics. Difference between means were calculated with Student's single sample t-test. RESULTS: Weight increased in all the ages, with significant differences in males at 7, 8 and 9 years (p<0.001) and in females after the age of 6 years (p< 0.05). Height significantly increased at all ages and in both sexes, except in females at the age of 5 years. Body mass index increased only at the ages of 8 and 9 years in males (p<0.01). Skinfold thicknesses significantly increased in both sexes and at all ages (p<0.05). Body density decreased (p<0.01) and body fat increased, both in kilograms and in percentage (p<0.01). CONCLUSIONS: During the 15 years studied, height and weight increased significantly. Body mass index did not increase significantly, whereas body fat (skinfold thicknesses and fat compartment) significantly increased. PMID- 11003878 TI - [Bone maturity evaluation by automatized metric measurements]. AB - OBJECTIVES: Precision and rapidity of computerised procedures facilitate bone maturity evaluation by measuring bone images. In this paper we analysed the adequacy of several of these measures to evaluate growth or bone maturity. MATERIAL AND METHODS: To determine growth or maturity indicating parameters, two hundred X-rays of the left hands and wrists of 2 to 18 years old children were examined by an automated image analysis procedure (IMAGO-2). Children were examined every six months (18 to 36 months) obtaining several auxological data (weight, height, bone maturity, TW2 methods, etc.). The relationship between the automated indicating parameters of maturity or growth and the auxological data were studied (linear correlation and multiple regression). RESULTS: All parameters correlate with the chronological age. The area parameter presents the highest coefficient of correlation with weight and Speed of Growth. The perimeter presents the best correlation with Height and bone maturity (TW2). Variations in some separation measurements from the circular ones relate well with auxological data changes. CONCLUSIONS: It is difficult to categorically catalogue one measurement as the only and exclusive indicator of maturity or growth. The studied parameters correlate best with bone maturity (TW2) when compared with those of the auxological data (weight, height) which could indicate their usefulness to evaluate bone maturity. PMID- 11003879 TI - [Opinion of heads of schools about the impact of school setting factors on the attitude of children and youngsters regarding smoking]. AB - AIM: Knowing the influence of school environment on children and teenagers' attitudes about tobacco consumption. METHODS: A survey was carried on 3.050 Spanish school headmasters (16% of the total in Spain), who were asked about the main variables related to the school environment able to influence the attitude of children and teenagers towards tobacco, such as the fulfilment of the official anti-tobacco legislation at educational centres, the education for nicotine poisoning prevention, or the teachers attitudes about tobacco, among others. Multiple regressions were made so as to value the variables that most influence the pupils' tasting tobacco and becoming usual smokers. RESULTS: In the opinion of the headmasters the variables that individually most influence the percentage of pupils that have tasted tobacco are the percentage of smoker teachers (beta 0.21) and the percentage of teachers who smoke in the presence of pupils (beta 0.091), as on the percentage of students usual smokers (beta 0.21 and 0.19 respectively). All the variables from school environment analysed jointly, explain some percentages of variance of the 9.65% and 10.7% of the estimated percentages of pupils who have tasted tobacco and pupils that are usual smokers, respectively. CONCLUSIONS: The efforts most be doubled, in order to reach the real fulfilment of the official anti-tobacco legislation in all educational centres and the pupils receive teachings for the prevention of tabaquisme in the context of Health Education, since these measures may make the schools play a more determinant role in the prevention of the nicotinism. PMID- 11003880 TI - [Clinical manifestations and biological markers in the natural history of HIV-1 infection in vertically infected children. Longitudinal study]. AB - OBJECTIVE: To study the relationship among clinical symptoms and biological markers as predictive value of progression to death in HIV-1 vertically infected infants. PATIENT AND METHODS: We carry out a prospective study in 43 HIV-1 infants with a mean age of 4.27 (range: 0-11.8 months). None of the infants' mothers had received any antiviral treatment during pregnancy. None of the infants were breastfed. They were routinely assessed for clinical symptoms during follow-up. RESULTS: Cox regression analysis was used to study the hazard ratio (HR) of progression to death. For the median viral load > 5 log10, the HR was 6.42 (95% CI, 1.28-32.03) (p = 0.023) and 6.84 (95% CI, 1.52-30,69) (p = 0.012) for biological phenotype of viral isolates with rapid replication and high titter (R/H-X4). We also study the predictive value of the clinical symptoms and we observe that the symptoms with more HR of progression to death were the progressive encephalopathy (3.60 [95% CI, 0.92-14.06; p = 0.065]) and the cardiopathy (6.29 [95% CI, 1.59-24.85; p = 0.008]). CONCLUSIONS: Our data indicate that viral load > 5 log10 and biological phenotype R/H-X4 of virus isolates along the study are predictive markers of progression to death. In addition, the progressive encephalopathy and cardiopathy were also markers of progression death. PMID- 11003881 TI - [Antibiotic prescribing patterns for pediatric inpatients with acute respiratory tract infection]. AB - BACKGROUND: Children with acute respiratory tract infection (ARTI) represent an important target group for efforts aimed at reducing unnecessary antibiotic use. OBJECTIVE: To present the epidemiological data and evaluate the effect of clinical, laboratory, radiological and microbiological data on the decision to prescribe antibiotics to pediatric patients with ARTI as well as to seek criteria that would justify antibiotic use. PATIENTS AND METHODS: A retrospective review was made of the clinical histories of 147 previously healthy children, consecutively admitted to our hospital with ARTI for 1 year (May 1996-April 1997). Patients were divided in two groups: those not treated with antibiotics (n = 92) and those treated (n = 55). Data from the two groups were compared with a statistical computer program (R-Sigma). RESULTS: Of the 147 patients studied, mean age was 2.5 years (range 0-14 years) and 85 (58%) males. One-hundred-and five patients (72%) had previously been attended to in the emergency room, and 45 patients (30%) had been treated with antibiotics. Upper respiratory tract infection was diagnosed in 81 patients (54%), bronchitis in 28 (18%), bronchiolitis in 23 (15%) and pneumonia in 15 (10%). Ninety-seven patients (66%) had viral infection and only two (1%) had bacterial infection. Syncytial respiratory virus was isolated in 41 patients (28%) and adenovirus in 30 (20%). In the untreated group, the longer duration of symptoms before admission, lymphocytosis, clinical diagnosis of bronchiolitis and normal thorax X-ray, were statistically significant. In the treated group, fever, leukocytosis, neutrophilia and a diagnosis of pneumonia were statistically significant. Length of stay was longer in this group than in the untreated group. CONCLUSIONS: It is difficult to prescribe antibiotics on the basis of bacteriologic data. Laboratory, analytic and radiological data can be helpful in the rational use of antibiotics. PMID- 11003882 TI - [Antibiotic prescribing patterns in primary health care. Do pediatricians use antibiotics rationally?]. AB - OBJECTIVES: To determine antibiotic prescribing patterns in the pediatric (infants and children) population attended to at a primary health care centre in the community of Madrid. We also wanted to determine the necessity or otherwise of antibiotic therapy and whether the selected antibiotic drug was appropriate for the pathology diagnosed. METHODS: Retrospective study of all infectious or respiratory processes diagnosed during 1 year and of the respective antibiotic cycles prescribed in all patients under the age of 4 years. The prescribing physician and the appropriateness of all therapeutic decisions, including those where the decision was not to treat with antibiotic drugs, were analyzed. RESULTS: We evaluated 910 children under the age of 4 years with a total of 3, 847 processes (mean of 4.55 +/-3.6 processes per child per year). Sixty-three percent of the children received at least one cycle of antibiotic drugs per year (mean 1.63+/-1.69 cycles of treatment per child per year). Of all therapeutic decisions, 85.2% were considered appropriate. In 36% of the processes antibiotics were prescribed (1,386 cycles), 46% of which were considered inappropriate either because no antibiotic therapy should have been given (71.6%) or because the chosen drug was not appropriate for the pathology (28.4%). There were significant differences among the evaluated physicians. The most correct decisions were taken by the pediatrician in the outpatient clinic, especially when compared with physicians in the emergency ward (p<0.0001). The most frequently prescribed antibiotic drugs were amoxicillin (41.2%) and amoxicillin combined with clavulanic acid (33%). Cephalosporin accounted only for 6.9% of the prescriptions. CONCLUSIONS: Antibiotic therapy is overprescribed in children, a situation that should be corrected. PMID- 11003883 TI - [Partial seizures in a newborn with tuberous sclerosis]. AB - Tuberous sclerosis is characterized by the potential for hamartomatous growth in multiple organs. Common manifestations are hypomelanotic spots, facial angiofibromas, subependymal hamartomas, cortical tubers, cardiac rhabdomyomas, retinal hamartomas, and so on. Seizures and mental retardation are frequent. It is an autosomal dominant disease but there is a high percentage of spontaneous mutations. Neonatal diagnosis is exceptional. We report a case of a female term newborn who presented partial motor seizures at the third day of life. Physical examination revealed only a disturbance of cardiac rhythm. Echocardiography showed ventricular intramural rhabdomyomas. Magnetic resonance imaging disclosed periventricular subependymal nodules and cortical tubers. A retinal hamartoma was found in the right eye. At the age of 1 month, hypomelanotic spots were evident on the back skin. The patient had infantile spasms, followed by poorly controlled partial complex seizures together with severe psychomotor retardation. Examination of both parents was normal. We discuss the uncommon diagnosis of tuberous sclerosis during the neonatal period, as well as the exceptional mode of presentation of our patient, with seizures in the early neonatal period, a phenomenon rarely reported in the literature. PMID- 11003884 TI - [Partial trisomy 1q due to maternal translocation]. PMID- 11003885 TI - [Neonatal hyperthyroidism]. PMID- 11003886 TI - [Chilaiditi's syndrome]. PMID- 11003887 TI - [Thanatophoric dwarfism in twin pregnancy]. PMID- 11003888 TI - [Bilateral coxalgia in a girl with Down syndrome]. PMID- 11003889 TI - [Detection of abnormalities in carbohydrate metabolism in cystic fibrosis]. PMID- 11003890 TI - [Detection of abnormalities in carbohydrate metabolism in cystic fibrosis] [In Process Citation] PMID- 11003891 TI - [Non-invasive mechanical ventilation for congenital central alveolar hypoventilation]. PMID- 11003892 TI - [Endopleural urokinase: adverse reaction]. PMID- 11003893 TI - [Remission of nocturnal enuresis in three patients with orthodontic advance mandibular appliance]. PMID- 11003894 TI - [Use of albumin in the critically ill and evidence-based medicine]. PMID- 11003895 TI - [Pneumococcal vaccines. One solution to several problems?]. PMID- 11003896 TI - [Rational use of antibiotics in pediatrics: impact of a rapid test for detection of beta-haemolytic group A streptococci in acute pharyngotonsillitis]. AB - OBJECTIVES: To assess the reliability and validity of a rapid test for the identification of Streptococcus pyogenes in the pharyngeal exudate of children presenting with pharyngotonsillitis. To evaluate the impact of its use in outpatient clinics on antibiotic use, on the incidence of second medical visits and complications, and on the degree of parental satisfaction. PATIENTS AND METHODS: After a clinical diagnosis of acute pharyngitis was established and written informed consent obtained from the parents, dual throat swabs were collected from 430 children who attended the emergency department of our hospital or the pediatric offices of three health centers in our area. The first specimen was examined by the rapid test, QuickVue Flex Strep A, and the second one was sent to the laboratory for conventional culture. As a rule, antibiotics were indicated only when the rapid test was positive. Special emphasis was placed on explaining to parents that treatment was not necessary when the test was negative. Telephone follow-up was provided to the family during the next four weeks, after which a satisfaction survey was carried out. RESULTS: The sensitivity of the investigated rapid test was 91.2% (negative predictive value: 96.5%) and specificity was 96.2% (positive predictive value: 90.4%). Antibiotics were given to 41.9% of the patients, approximately half the expected rate in the absence of the rapid test. There was no significant difference in the number of second visits or hospitalizations between the groups of treated and non-treated subjects. Clinical evolution was good in all cases. The degree of parental satisfaction was very high, independent of the treatment given to the patients. CONCLUSIONS: The rapid test for the detection of group A streptococci is a reliable tool for the selection of patients able to benefit from antibiotic treatment. It is easy to handle and apply and its use allows a significant reduction in the administration of antibiotics in pharyngotonsillitis. Most users accept and are satisfied with this novel diagnostic and therapeutic procedure. PMID- 11003897 TI - [Immunogenicity of the 23-valent pneumococcal vaccine in children with chronic renal disease]. AB - OBJECTIVE: To study the immunogenicity of the 23-valent pneumoccal vaccine in children with chronic renal disease and in those with nephrotic syndrome, and to compare it with the response in healthy children. METHODS: The vaccine was administered to 150 children aged 2-18 years: 113 with renal diseases (26 with nephrotic syndrome, 23 with severe grades of vesicoureteral reflux, 16 with chronic renal insufficiency, 6 renal transplant recipients, 6 requiring dialysis and 36 with other renal diseases) (group 1) and 37 healthy (group 2). Specific IgG antibodies concentrations were measured by ELISA before and 30 days after vaccination. The results obtained in both groups were compared. We compared too the response observed between the subgroup of children with renal diseases in which pneumococcal vaccine is indicated (nephrotic syndrome, chronic renal insufficiency, renal transplant and those requiring dialysis (group 3) and healthy children. RESULTS: 87.3% of children showed a 2-fold increase in antibody concentrations after vaccination. No significant differences were observed between the three groups. We considered that the vaccine was immunogenic in 78.4% of healthy children, in 77.9% of group 1 children and in 72.5% of those in the group 3 (p = 0,533). A lower response was observed in children with a kidney transplant and in those requiring dialysis. CONCLUSIONS: These results suggest that 23-valent pneumococcal vaccine is immunogenic in children with chronic renal diseases or nephrotic syndrome and may protect these patients from invasive pneumococcal disease. The importance of improved vaccine coverage is emphasized. PMID- 11003898 TI - [Hospital guidelines on breastfeeding in the north-center of Spain]. AB - OBJECTIVES: To investigate the influence of the maternity protocols on frequency and prevalence of breastfeeding in the 15 most important maternity units from the north-center region of Spain. METHODS: From October 1992 to December 1992 we collected 1175 newborn babies and we followed them up for 12 months. A protocol study was designed to collect the data in DBase IV format and the statistical study was done with SPSS version Window 6.0. chi 2 test was applied. RESULTS: The presence of the father at the delivery did not influence breastfeeding. The rooming-in was an adverse factor. Instrumental delivery and specially cesarean section act as an adverse factor for breastfeeding. To feed the baby with glucose or water and to delay the breast exposure was also negative to breastfeeding. CONCLUSIONS: The protocols used in the Spanish north-center maternity units are at a distance from those of Baby-friendly-Hospital Initiative. The early exposure to the breast and the restriction of bottle-feed with glucose in the early hours of life favours breast feeding. Instrumental delivery act as an adverse factor for breastfeeding. PMID- 11003899 TI - [Tuberculous meningitis: a disease in regression in our country?]. AB - OBJECTIVE: Our aim was to analyse clinical, diagnostic, therapeutical and evolutionary features in a pediatric population with tuberculous meningitis. PATIENTS AND METHODS: The medical records of thirteen children with this diagnosis admitted to Hospital Infantil Virgen del Rocio from Seville (Spain) between 1984 and 1999 were reviewed. RESULTS: The mean age was 2,35 +/- 2,3 years. The symptoms upon admission were: fever in 11 children, anorexia and vomiting in 8, disturbance of the consciousness in 7. Meningeal signs in 6, all of them older than 20 months, the remaining seven showed irritability and four of these ones hypertense fontanelles. Three patients were in the first stage of the disease, 9 in the second and 1 in the third, according to the Medical Research Council. CSF findings were indicative in all the cases. Five children had bacilloscopy positive and Mycobacterium tuberculosis was isolated in 6 patients, sometimes in CSF others in gastric juice. Mantoux skin test was positive in 11. Radiographic studies demonstrated abnormal chest findings in 8 patients (hiliar adenopathy, 1; miliary pattern, 2; and infiltrates, 5). Pathology cranial computed tomography showed in all the cases and the electroencephalogram was slowed down in the initial phases in 11. Two children died and the neurological complications were the most frequent, appearing in 9 patients. Without consequences cured 4 patients, the rest presented cognitive, visual and motor deficits, sensibility skin disturbance and late seizures. No case has been observed during the last 5 years. CONCLUSIONS: Fast diagnosis tests used for M. tuberculosis identification were useful to begin an antituberculous treatment in a high suspicion of meningeal affectation by this German patient. The early treatment will decrease complications and consequences by this disease. A decrease in the incidence looks to be in spite of the VIH infection increase nowadays. PMID- 11003900 TI - [Ureaplasma urealyticum and pertussis-like syndrome]. AB - BACKGROUND: Ureaplasma urealyticum is associated with respiratory pathology in the neonates and preterm neonates. However, this association has been poorly studied in infants and during early infancy. To address this issue, a clinic evaluation of patients with whooping cough and isolation of U. urealyticum in their nasopharyngeal aspirates has been done. METHODS: Over a period of 11 years, 1063 nasopharyngeal aspirates from 905 infants were studied. Clinical samples were cultured for Bordetella spp., other bacteria, viruses and mycoplasma. Data of patients with positive cultures for U. urealyticum were obtained from clinical records. RESULT: U. urealyticum was isolated from 26 patients with a median age of 5 months (range: 23 days-22 months). The gestational age of 9 patients (34.6%) was less than 37 weeks. All the patients were hospitalised because of pertussis like syndrome, which was associated with respiratory distress due to bronchospasm in 18 patients (69.2%). Twelve patients (46.1%) had fever and 15 (57.7%) showed lymphocytosis. The chest roentgenogram was abnormal in 18 patients (69.2%): pulmonary hyperaeration, with or without atelectasis. Clinical evolution was good in all patients. In 16 patients (61.5%) U. urealyticum was isolated together with other microorganisms: in 9 samples with bacteria (H. influenzae, S. pneumoniae, B. pertussis, M. catarrhalis), in 5 with viruses (respiratory syncytial virus, cytomegalovirus, adenovirus, enterovirus) and in 2 samples with respiratory syncytial virus and S. pneumoniae and B. pertussis respectively. CONCLUSIONS: Likely U. urealyticum cannot be considered clearly as the etiologic agent of whooping cough, mainly because in the 61.5% of patients U. urealyticum has been isolated together with other microorganisms considered pathogens or potentially pathogens. Future studies would be necessary in order to establish the pathogenic role of U. urealyticum after the neonatal period. PMID- 11003901 TI - [Analysis of treatment failure in pediatric subglottic stenosis]. AB - BACKGROUND: The good results obtained until nowadays in the surgery of subglottic stenosis is followed by an analysis of the failures. METHODS: We study retrospectively 48 cases treated in the last eight years in two institutions. Study variables were: age, previous diagnosis, grade os the stenosis, surgical treatment, complications and evolution. RESULTS: Mean age was 2.8 +/- 0.43 years (newborn-17 years). In 13 cases (27%) the result obtained was considered a failure. Failure indexes were: treatment selection (7,7%), surgical technique selection (23%), due to the technique itself (23%), iatrogenia (15,4%), associated diseases (30,7%). Mortality of the series was 8.3%. CONCLUSIONS: The learning curve of these techniques is difficult although once the necessary experience is achieved the results are excellent. PMID- 11003902 TI - [Experience with ventriculitis at a neonatology department]. AB - OBJECTIVE: To study the characteristics, treatment and follow-up of patients with ventriculitis in our neonatal unit. PATIENTS AND METHODS: Retrospective study of patients diagnosed with ventriculitis from January 1990 to December 1997. Diagnostic criteria were the identification of any bacteria in the ventricular fluid and pleocytosis (> or = 100 leukocytes per microl). Personal history, clinical and analytical findings and evolution after diagnosis were studied. RESULTS: We recorded ten cases of ventriculitis in nine patients. Six of them occurred as a complication of previous meningitis and four occurred after neurosurgical treatment. The mean age at diagnosis was of 38.8 days (range 8 130), and mean gestational age was 29.4 weeks (range 25-38). Clinical and ventricular fluid anomalies were seen in six cases and in four the diagnosis was made at autopsy. Treatment was systemic antibiotics. In two cases intraventricular antibiotics were added. Six patients died, ventriculitis being the direct cause of death in five. Of the three survivors, one had mental retardation and cerebral palsy and the other two had minor disabilities. CONCLUSIONS: During the neonatal period, a high degree of clinical suspicion and techniques for an early diagnosis and treatment are needed for ventriculitis. PMID- 11003903 TI - [Oxidative phosphorylation defects with neonatal presentation: review of our caseload]. AB - OBJECTIVES: To define the oxidative phosporilation deficit syndrome in the neonatal in terms of incidence and clinical, biochemical and genetic features. MATERIAL AND METHODS: We report 9 newborns diagnosed as oxidatic phosporilation deficit during the last 8 years in our hospital by means of clinical, metabolic, pathological and molecular studies, among other evaluations. The diagnosis was established based on ensymatic deficit of the respiratory chain, associated with alterations in the mtDNA in one case, and with mitochondrial ultrastructural anomalies in 5 cases. RESULTS: There was an incidence of 1/3.555 newborns and 1/832 newborns admitted in our Neonatal Unit. In four of them there were familial antecedents and polihidramnios in two. Most of them, 8 out of 9, were born at term after a normal pregnancy and delivery, with normal Apgar score and auxological examination. Symptomatology started immediately at the neonatal period as acute neurological damage in most of them. There was a severe evolution as 5 children died and 4 survived with severe damage. All of them had the classical phenotype of early severe encefalopathy, associated with dismorphic features, hypotomia, neurosensorial defects, brain dysgenesis and atrophy, anomalies in the EEG and in 5 of them there were also systemic anomalies, mainly cardiopathy. The most frequent biochemical alteration was a significative increment of the quotient lactate/piruvate. Five patients presented ultrastructural alterations of the mitochondria in thr muscle biopsy but Cox stain was not positive in any case. Three cases has a deficit of the complex IV, e of the complex I-IV, 2 of the complex I and one the complex I-III-IV. Only one patient had multiple deletions in the mtDNA. CONCLUSIONS: Oxidatic phosporilation deficit are frequent and severe diseases of prenatal onset with limited fetal effects, homogeneous clinical phenotype with frequent damage of the central nervous system and variable extraneurological alteration and inconsistent biochemical pattern. Enzymatic studies ar need for making the diagnosis in all suspected cases, PMID- 11003904 TI - [Adolescents with growth retardation]. PMID- 11003905 TI - [Toxic shock syndrome in children. Report of four cases]. AB - We report four children with toxic shock syndrome admitted in the pediatric intensive care unit of our hospital during the past year. All the children had the five criteria established by the Centers for Disease Control for the diagnosis of this syndrome. In all four there was a probable point of entry of the infection: maxillar sinusitis in one, pneumonia in two and surgical wound in the other. No bacteria that could have caused the infection were isolated in any of the children, which suggests a staphylococcal origin for this syndrome. Evolution was good in all of the children due to aggressive treatment that included inotropic support, volemic expansion and antibiotics. Two of the children, who suffered adult respiratory distress syndrome, required prolonged respiratory support. PMID- 11003906 TI - [Isolated hyperferritinemia in a healthy male infant: hereditary hyperferritinemia-cataract syndrome]. AB - The case of a male infant who was found to have hyperferritinemia was made at the age of 3 months is described. The patient and several members of his family from three generations were diagnosed with hereditary hyperferritinemia-cataract syndrome with a new point mutation in the iron-responsive element of the L ferritin gene. Differential diagnosis of hyperferritinemia is discussed with emphasis on the need for the knowledge of this entity to avoid unnecessary investigations. PMID- 11003907 TI - [Traumatic pulmonary pseudocyst]. AB - Traumatic pulmonary pseudocyst is an unusual complication that appears after a closed thoracic trauma. It is produced as a consequence of outburst and shear forces released by the impact on the elastic thoracic wall. We present a 14-year old boy who, after a motorcycle crash, presented a traumatic pulmonary pseudocyst, isolated on the right hemithorax. The patient's evolution was good. The antecedent of trauma, together with radiological examination and the tendency toward spontaneous resolution, suggested the diagnosis as well as the advisability of a conservative approach in most pediatric cases. Recognition of this unusual disorder would help to avoid unnecessary diagnostic and therapeutic procedures. PMID- 11003908 TI - [Acute interstitial nephritis of probable pharmacological origin]. AB - Acute interstitial nephritis in children is rare. We present a case of acute interstitial nephritis in a 10-year-old boy, which was probably drug-induced. Initial symptoms included fever, loss of appetite, weight loss, alterations in urine analysis and mild renal failure. Treatment with steroids produced a good clinical response and renal function returned to normal within a few months. PMID- 11003909 TI - [Focal nodular hepatic hyperplasia. A report of a new case]. AB - Focal nodular hyperplasia is a benign liver tumor, that is rare during childhood. We report a new case, exceptional by the short age of the patient, and by the way of this we review the epidemiology, the actual diagnosis criterion and the treatment options. PMID- 11003910 TI - [Use of intrapleural urokinase in children]. AB - OBJECTIVE: To present the use of intrapleural urokinase as an alternative to surgical management of complicated pleural effusions in children. To add new cases to the literature and set bases for the development of a standardized protocol. METHOD: Presentation of three cases in children under three years of age treated with intrapleural urokinase. RESULTS: Resolution of the effusions with lack of adverse effects. CONCLUSION: Treatment of pleural effusions when loculations develop. This invasive approach might be avoided in some cases by instillation of a fibrinolytic agent such as urokinase into the pleural space, this being an efficient and safe procedure. PMID- 11003911 TI - [Phenotype heterogeneity in Gitelman's syndrome]. AB - A family with three offspring affected of Giltelman's syndrome is reported. The phenotypic variability of this entity is emphasized. Moreover, the diagnosis criteria of the syndrome, phathophysiology, and genetics and clinical differences with Bartter's syndrome are stated. PMID- 11003912 TI - [Cohen's syndrome: non-causal association with vascular rings]. AB - Cohen syndrome is an autosomal recessive disorder characterized by hypotonia, mental retardation, microcephalia, typical craniofacial features, myopia and chorioretinal dystrophy. The responsible gene has been mapped to chromosome 8q 22 (COH 1). Since it was described more than 100 patients have been reported. However, none of them has been associated with vascular rings. Our hospital has studied eight pediatric cases and 25% of them were related with vascular rings. PMID- 11003913 TI - [Use of the International Classification of Epilepsies and Epileptic Syndromes in epidemiologic studies]. PMID- 11003914 TI - [International classification of the epilepsies and epileptic syndromes in epidemiologic studies] [In Process Citation] PMID- 11003915 TI - [Erythema nodosum associated with Salmonella enteritidis infection]. PMID- 11003917 TI - [ [In Process Citation] PMID- 11003916 TI - [Neonatal papular acrodermatitis (Gianotti-Crosti) and Bartonella henselae infection]. PMID- 11003918 TI - [Imported malaria in children]. PMID- 11003919 TI - [Imported malaria in children (1993-1998)]. AB - Malaria is the world's most important parasitic infection, accounting for an estimated 300 million cases and 1.5 to 3 million deaths annually. Although it has been eradicated from temperate countries, increasing numbers of travellers visit each year tropical countries, where malaria remains a major cause of morbidity and mortality. Children are particularly at risk from malaria since symptoms can be especially severe and can develop rapidly. Symptoms may differ from those in adults and, as children often have febrile illnesses, malaria could not be suspected. Eleven cases of imported mala-na in children which ages was between 15 month until 13 years, were seen in the Sant Joan de D|u Hospital, Barcelona (Spain) since August 1993 until August 1998. The aetiology was in 7 cases Plasmodium falciparum and in 4 cases Plasmodium vivax, in one case we found also with the P. falciparum some forms of P. malariae. All of them experience fever and anaemia at admission and nine had splenomegaly; the evolution after the treatment was fine. Cases of imported malaria in children are increasing and it is timely to review the prevention and the management. PMID- 11003920 TI - [Surgical treatment of supravalvular aortic stenosis]. AB - OBJECTIVE: Surgical correction for 10 supravalvular aortic stenosis since 1988. MATERIALS AND METHODS: Diagnosis was carried out by means of echocardiography and magnetic resonance. Seven patients showed features of Williams-Beuren syndrome, 5 patients showed in their genotype a delection of 7-chromosome. 5 showed membranous-localized type obstruction and the other 5 with hourglass type. The patients in the first group underwent circumferential resection of the stenosis ring followed by a diamond- shaped patch and the second group were treated with resection of the stenosing ring associated with an inverted Y- shaped patch with releasing of the coronary ostia in two of them. RESULTS: There was no hospital death. The gradient was substantially reduced from 60 +/- 8 to 5 +/- 1 mm Hg (range 0-14). The average staying was under 7 +/- 1 days. The average follow-up was 58 +/- 8 months (2-120). CONCLUSIONS: Postoperatively, all the patients were in NYHA class I-II. The residual gradient was less than 15 mmHg in the echocardiography study during the follow-up and none of them required an additional operation. PMID- 11003921 TI - [Fisher's index as indicator of severity and its course in the postoperative period following liver transplantation in children]. AB - OBJECTIVES: To study the evolution of Fisher index during the early postoperative period after liver transplantation in children. PATIENTS AND METHODS: We have studied 32 liver transplants performed on 26 pediatric patients, during the first week of the post-operative period. In all cases the BCAA/AAA quotient was determined before transplant, at the time of PICU admission, and on days 1, 2, 3, 4, 5, 6 and 7 after transplant. RESULTS: Compared to control group values, the Fisher index for these patients is significantly lower at pre-transplant (p < 0,0001), upon admission (p < 0,001), and days 1 (p < 0,0001), 2 (p < 0,0001) and 3 (p < 0, 0001). Comparison between non-survivors and survivors shows a significant reduction in the first group on the index in days 1 (p < 0,02), 3 (p < 0,02), 4 (p < 0,005), 5 (p < 0,002), 6 (p < 0,03) and 7 (p < 0,01) of post transplant. CONCLUSIONS: The evolution of the Fisher index can be useful as an indication of the severity of the condition of these kinds of patients, which would help to establish more aggressive treatment to improve prognosis. PMID- 11003922 TI - [Bone mineral density and bone mineral content variations in various communities]. AB - OBJECTIVE: To study the bone mineral density (DMO) and the bone mineral concentration (CMO) in lumbar spine (L2 and L4) by dual-energy x-ray absorptiometry with a lunar DPX (DEXA) in a children sample of the community of Madrid; to relate the values obtained with the age, sex and pubertal development; and to compare the values of DMO found with the publications of other autonomous communities. MATERIALS AND METHODS: 351 children, 184 boys and 167 girls selected at random in our environment. The age range oscillated between 6 months and 20 years. Grouped in intervals of a year according to the sex; and in accordance with the pubertal development. The bone mineral content was measured by dual energy x-ray absorptiometry with a lunar DPX in the lumbar spine at level of L2 L4. The statistic analysis has been accomplished with the SPSS version 6.0.1. They have been obtained the average and deviation standard for each group from studied age, and also according to pubertal development; the DMO and CMO of the boys and girls have been compared by groups of age; the DMO and CMO within each sex between a group of age and the immediately superior have been compared through an analysis of the variance; the effect that the age has on the DMO and the CMO has been evaluated and finally through multivariant analysis techniques the regression models have been estimated between the age and the DMO, and the age with the CMO. CONCLUSION: The DMO shows variations between the various communities and even in different samples of a same community, in Carrascosa study the DMO is highest, expressed in SD score, in many several groups of age studied with respect to our values and with the values of Moreno et al and Armada et al; the values of DMO in our study are greater than the ones found by Moreno et al and Armada et al; the first four years and the adolescence are the periods of maximum increase of the DMO, but it also increases in an oscillatory way in the intermediate stages; the girls present some highest levels of DMO in the groups of age of 12-13 and 14-15 years, probably in relationship to a most precocious beginning of the puberty; and finally the regression line of the DMO as compared to the age for both sexes are parallel and have equal court point, what means that they are coincident. PMID- 11003923 TI - [Peak expiratory flow rate in 4- to 15-year-old children. Comparison of 3 measuring models]. AB - OBJECTIVE: a) To establish the possible differences between three different models of PEF-meter, including PF-Control for the first time; b) to build a normogram for each of the three models, as a function of different variables (age, height, and inspiratory and espiratory thoracic circumference); c) to search for differences between both sexes, and d) to study the mathematical correlation between the results yielded by the different models. METHODS: PEF was measured in a sample of 476 school-children, in the range of 4 and 15 years (233 boys, 243 girls), with a mean age of 9,3 years and a height range between 96,8 cm and 178,8 cm (mean 137,9). PEF value was recorded as the best of three attempts when the difference them was less than 10%. Three devices of each model were used (Vitalograph, Mini-Wright, PF-Control). The utilisation was always done in a rotatory fashion at school time (15-17 hours) in the children's school. RESULTS: We found significant differences between the three models (p < 0,0004). The best predictive variable for PEF is height, with an r = 0,90-0,92 (depending on the model) when the exponential model is considered. Independently of the device, there were significant differences between sexes. CONCLUSIONS: a) The variable that expresses PEF the better is height; b) PEF values are different between girls and boys; c) there were significant differences between the various models; d) Monitoring of a given child must be done with the same model of PEF-meter, and e) The best correlation is obtained with the exponential model, although differences with the lineal model were not statistically significant. PMID- 11003924 TI - [Insulin Lispro (Lys B28, Pro B29) treatment in adolescents and young people with type 1 diabetes]. AB - BACKGROUND: The human insulin analogue, Lispro (Lys B28, Pro B29), is more similar to normal pancreatic insulin response to ingestion. Therefore, it could provide an alternative to the classical treatment of type-1 diabetes in childhood. The aim of this study was to analyze the response to insulin treatment with this analogue during 1 year. PATIENTS AND METHODS: In a study group of twenty puberal diabetic patients (nine male and 11 female) with a mean age of 15.6 years (+/- SD) and with diabetes of a mean of 8.3 years (+/- 4.3 SD), classical intensive treatment, a combination of regular insulin (HR) and NPH insulin (Humulin NPH) was substituted for a new treatment with Lispro and NPH insulin in 3-4 doses. All patients had received diabetic education and performed at least four blood glucose tests daily and self monitoring. We analyzed the 6 months prior to the change in treatment (Group A) and the 12 months after the change (Group B). The amount of insulin (u/kg/day), the fast/intermediate insulin ratio in each dose, dietary modifications, the level of metabolic control given by the HbA1C average measured every 2 months, severe hypoglycemia and the patient s level of satisfaction with the new treatment were compared between the two groups. RESULTS: The number of daily doses, as well as the daily insulin intake, was the same in both groups (3.7 [+/- 0.6] doses/day; 0.9 [+/- 0.2] u/kg/day). With Lispro treatment the ratio fast/intermediate insulin was reduced. This reduction was statistically significant for the pre-breakfast dose (Classical = 65. 4 +/- 30% vs Lispro = 47.1 +/- 19.6%), and for the pre-lunch dose (Classical = 58.1 +/- 29.3% vs Lispro = 39 +/- 12.8%). Most patients did not need neither mid-morning or mid-afternoon doses. There were no modifications in body mass index. Although metabolic control improved slightly (Classical X A(b) A(1)c = 7 +/- 1.2 vs Lispro X A(b) A(1c) = 6.6 +/- 1.1), the difference was not statistically significant. Three patients had a severe hypoglycemic episode in the first 6 months with the new treatment pattern. All of them were satisfied with the new insulin. CONCLUSIONS: Intensive therapy with Lispro insulin combined with appropriate doses of basal insulin (NPH) can provide a good alternative in the treatment of diabetic teenagers. PMID- 11003925 TI - [Use of a hospital pediatric emergency department during the night]. AB - OBJECTIVE: To determine the characteristics of children younger than 14 years visiting our pediatric emergency department between 0:00 and 8:00 hours. PATIENTS AND METHODS: Retrospective review of 300 children visiting our emergency department at night. RESULTS: Between March 1 and 22 1999 we recorded 300 episodes from 0:00-8:00 hours (1.7 patients/hour) and 2350 episodes from 8:00 24:00 hours (6. 6 patients/hour). Of the 300 episodes registered at night, 132 children (44%) came between 0:00 and 2:00. The most common complaints were: respiratory symptoms in 116 patients (38.6%), fever in 61 (20.3%) and digestive symptoms in 61 (20.3%). We carried out at least one complementary test in 111 patients (37.0%). Five children (1.7%) were admitted to the hospital (80 between 8:00 and 24:00, 3.4%, p = 0.15) and 25 (8.3%) were admitted for a few hours to the observation ward (123 of the 2350 who came between 8:00 and 24:00, 5.2%, p = 0.06). Final diagnoses were: ear nose and throat infection in 91 (30.3%), fever without source in 38 (12.6%), asthma in 29 (9.6%), acute gastroenteritis in 27 (9%), croup in 22 (7.3%), vomiting in 14 (4.6%), abdominal pain in 13 (4.3%), febrile convulsion in 6 (2%), pneumonia in 5 (1.6%), bronchiolitis in 5 (1. 6%), bacteremia in 1 (0.3%), and other diagnoses in 46 (16.3%). CONCLUSIONS: The number of visits to our emergency department diminished at night, particularly between 0:00 and 2:00. The night-time admission rate was less than the daytime rate, although this difference was not statistically significant. Admission for a few hours to the observation ward was more common at night. The percentage of patients with respiratory illnesses was higher during the night. PMID- 11003926 TI - [Survival at discharge of newborn infants weighing less than 1000 g (1991-1997). When are they too small?]. AB - OBJECTIVE: To describe survival rates at discharge in liveborn infants with birth weight under 1000 g by birth-weight and gestational age categories in a third level spanish hospital in order to improve health care planning and parental counselling about the threshold of viability. STUDY DESIGN: A follow-up study of 178 infants with birth-weight under 1000 g born alive between 1991 at 1997. Six newborns with lethal malformations were not included. RESULTS: Overall survival at discharge was 54%. During the study period survival rate improved in the birth weight group 750-999 g. In the group below 750 g survival rate was lower and improved more slowly. Survival rate for infants below 750 g and 26 weeks was extremely low. Over 749 g and 25 weeks survival was over 50%. CONCLUSIONS: Survival rate showed an improving trend during the study period. In the last years of the study period survival in infants with birth-weight over 749 g was above 85%. For infants under 750 g and 26 weeks was extremely low. Family and medical decision making about active treatment of these newborns should be an individualized process. PMID- 11003927 TI - [Limb reduction defects in Asturias (1986-1997): prevalence and clinical presentation]. AB - AIM: To determine the prevalence and clinical presentation of limb reduction defects in Asturias (Spain). PATIENTS AND METHODS: Among the 92300 newborns, stillbirths and induced abortions for congenital defects surveyed by the Registry of Congenital Defects in Asturias (population-based registry) during 1986-1997, we studied the children with limb reduction defects. Frequency is expressed as total prevalence per 10000 births. RESULTS: Seventy-three children with limb reduction defects were identified, with a total prevalence of 8 per 10000 births (CI 6.2-9.8): upper limbs were affected in 65.7%, lower limbs in 23.3% and both upper and lower limbs were affected in 11%. Transverse terminal defects were present in 49.3%, and were intercalary in 16.4%, preaxial longitudinal in 8.2%, postaxial longitudinal in 9.6%, split hand/foot in 9.6% and multiple in 6.8%. Limb deficiencies were found in 52%; of these, 48% were associated with other congenital defects. Four had chromosomal anomalies and 20 were syndromes. 31% had low birth weight for their gestational age. Alcohol exposure during pregnancy was recorded in three mothers. CONCLUSIONS: A detailed study of limb reduction defects could lead to a better understanding of clinical presentation and to an etio-pathogenic diagnosis to control risk factors. PMID- 11003928 TI - [Indications for anti-regurgitation formulas. Committee on Nutrition of the AEP]. PMID- 11003929 TI - [Facial and buttock dermatitis]. PMID- 11003930 TI - [A case of neonatal malaria in Spain]. AB - We report a case of neonatal malaria born in Spain. It is about a female newborn whose mother lived the first eight months of her pregnancy in Ecuatorial Guinea. Although our patient was well, in the third week of her life she developed fever mostly in mornings without any other symptoms except pallor. She kept a good physical state in any moment. In complementary proves we remark: anaemia and thrombocytopenia; as well Plasmodium falciparum ruin was found in blood smears. Treatment with mefloquine was successfully, blood smears was negative of parasites in the eighth day and hemogram was restoring normal. This article suggests neonatal malaria must be considered in those newborns suspected congenital infection born from mothers who have travelled to risk countries or immigrated from endemic areas. Also we remark that malaria clinic development in newborns is nonspecific and indistinguishable from other congenital infections. PMID- 11003931 TI - [Effectiveness of hemodynamic treatment guided by gastric intramucosal pH monitoring]. AB - OBJECTIVE: To determine whether gastric intramucosal pH can be used as an indicator for the treatment of critically ill children. METHODS: A protocol for hemodynamic treatment was tested in 34 critically ill children (medical and surgical patients). Endpoint was considered as normalization of gastric intramucosal pH. Hemodynamic complications were compared with another group of critically ill patients who received conventional treatment. RESULTS: Treatment designed to improve tissue oxygenation (hemodynamic stabilization, volume expansion, and dobutamine perfusion) raised pHi values from 7.25 +/- 0.04 to 7.32 +/- 0.13 (p = 0.014). Seventy-three percent of the children with pHi < 7.30 presented hemodynamic complications. These findings were compared with the results obtained in a control group of 30 critically-ill children given conventional treatment in the pediatric intensive care unit. In this group, 93.8% of the children with a pHi < 7.30 developed hemodynamic complications. CONCLUSIONS: We conclude that gastric intramucosal pHi can be useful as an indicator for the hemodynamic treatment of critically ill children. PMID- 11003932 TI - [Therapeutic efficacy of ketoconazole in Cushing syndrome]. PMID- 11003933 TI - [West's syndrome associated with calcified frontal dysplasia. Favorable treatment response]. PMID- 11003934 TI - [Cystic angiomatosis]. AB - A case of cystic angiomatosis is presented in a patient whose clinical sign were the presence of an inguinoescrotal mass due to a retroperitoneal lymphangioma, visceral splenic lesions and diffuse skeletal lesions. This rare entity is discussed and a brief review of the literature is made. PMID- 11003935 TI - [Severe neonatal mitochondrial cytopathy caused by isolated COX defect]. AB - We report a neonate with isolated cytochrome c oxidase (COX) defect and severe multisystemic involvement. The patient had severe encephalopathy, predominant since birth, and died due to hypoxic-ischemic myocardiopathy. He was the second son of non-consanguineous, healthy parents who also had a daughter with chronic encephalopathy. The neonate presented dysmorphic phenotype, hepatic and muscular involvement, and possibly tubular involvement. Metabolic studies revealed markedly increased lactic/pyruvic concentrations. Diagnosis was based on muscular enzymatic studies and ultrastructural mitochondrial anomalies, while the mitochondrial DNA and results of the COX technique were normal. Histological examination revealed a massive subendocardial infarction. Aspects of this entity with relevance for genetic counseling are discussed. PMID- 11003936 TI - [Median raphe cysts of the perineum in childhood]. AB - Median raphe cysts of the perineum are uncommon congenital lesions. Congenital lesions rarely encountered in daily clinical practice. Their diagnosis in childhood is particularly rare. We report two children with these cysts requiring surgical removal. We review the embryological, diagnostic and therapeutic aspects of these cysts, which could be of interest to pediatricians and pediatric surgeons when faced with this pathology in children. PMID- 11003937 TI - [Cervical spondylodiscitis in an infant]. AB - Spondylitis in childhood is rare, presenting a great variety of symptoms that are related to the child s age and to the site of the inflammatory process. We present the case of a nine-month-old infant, who presented to the emergency department with a torticollis of six weeks evolution. The diagnosis of spondylitis (C6-C7) was confirmed by MRI. Intravenous antibiotic therapy was administered. The patient showed complete regression of symptoms and imaging tests were normal. The rarity of this disease in patients in this age group, as well as the site and form of presentation are emphasised as is the importance of diagnosing this disease to prevent permanent neurological injuries or bone deformities. PMID- 11003938 TI - [Orofaciodigital syndrome associated with agenesis of the pituitary gland]. AB - Oral-facial digital syndrome (OFDS) consists of a group of heterogeneous genetic disorders with different patterns of inheritance. These disorders share facial, oral and digital abnormalities. At least 11 types of this syndrome have been described (I-IX) and the association between OFDS type II and absent pituitary has recently been reported in the case of two brothers. OBJECTIVE: 1. To describe the case of patients with OFDS associated with the absence of the hypophysis and panhypopituitarism. 2. To analyse whether the association between the agenesis of the hypophysis with OFDS represents a new type of this syndrome. PATIENT: An 11 month-old male infant presented with a malformation syndrome with oral (Gothic palate, lobulated tongue with lateral frenulae and hamartomas) facial (a small nose with flattened nasal root and hypoplastic nostrils, small facial skeleton, high forehead, low-implanted ears, stenosis of the choana) and digital malformations (poly- and clinodactyl hands and feet) typical of OFDS. In addition to these findings, at the age of 11 months he had growth retardation (length - 5.9 SD), obesity, mild psychomotor delay, a single upper incisor, micropenis and undescended testes. Endocrinologic studies showed combined GH, TSH, LH and FSH deficiency. Cranial magnetic resonance imaging showed a smooth sella turcica, lacking the pituitary gland, and the absence of the normal sign of neurohypophysis. Replacement therapy with GH and 1-thyroxine was started. SUMMARY: We report a patient with the clinical characteristics of OFDS type II, associated with the absence of hypophysis and panhypopituitarism. CONCLUSION: The infrequent association of agenesis of the pituitary gland and panhypopituitarism with OFDS, and the previous report of two brothers with this syndrome and the same association, suggests that this is a new type of OFDS. PMID- 11003939 TI - [Acute bulbar palsy due to myelomeningocele and Chiari II malformation]. AB - We describe a case of a 10-year-old girl with myelomeningocele and type II Chiari malformation. She presented with an acute bulbar palsy requiring mechanical ventilation. Surgical decompression of the brainstem was performed and, because of a phrenic nerve palsy, surgical plication of the right diaphragm was also required. The patient s evolution was favourable. We discuss the various forms of clinical presentation of this pathology as well as its management. The importance of early diagnosis and surgical correction are also stressed as a key factor for favourable evolution. PMID- 11003940 TI - [Renal biopsy and kidney diseases]. PMID- 11003941 TI - [Indications for renal biopsy in idiopathic nephrotic syndrome in children. Results of a national survey]. AB - BACKGROUND: The role of the renal biopsy in children with nephrotic syndrome is controversial, especially in patients with frequent relapses or corticosteroid dependent nephrotic syndrome. METHODS: We have sent a survey about the nephrotic syndrome to all the Spanish pediatric nephrologists who are members of the Spanish Association of Pediatric Nephrology. The survey is based in 7 questions about biopsy utility in the different circumstances of the illness. RESULTS: The survey reveals a scarce participation, 50%. The 85% of the inquired people uses the same diagnostical criterion. Referring the indication of renal biopsy in this disease, the 85% considers that it would be necessary to do it in case of corticosteroid resistance. The 57.7% of pediatric nephrologists would recommend a biopsy in children with frequently relapsing or corticosteroid-dependent nephrotic syndrome prior to cytotoxic therapy. The 33% doesn't practice biopsy before cytostatic administration, only when alterations in the renal function appear during the treatment. The 50% of the inquired considers that less than a year of age is an indication of biopsy in the nephrotic syndrome, and the 24.4% thinks that the age should never be an indication. CONCLUSIONS: The surveys aren't much used in our country as a resource of information. We think that the participation of expert people in the surveys gives validity to the obtained results. There are different opinions among pediatric nephrologists concerning the biopsy in children with frequently relapsing and corticosteroid-dependent nephrotic syndrome. PMID- 11003942 TI - [Burnout among Spanish pediatricians specialized in intensive care]. AB - BACKGROUND: The burnout syndrome is characterized by emotional exhaustion, depersonalization and decrease or loss of the feeling of personal accomplishment. It is frequent in the helping professions where an intense involvement with people needing assistance occurs. In its origin, the work policy factors seem to be important. METHODS: 68 pediatric intensive care physicians of the Intensive Care Section of the Spanish Pediatric Association participate in the study. A general questionnaire and a Spanish version of the Maslach Burnout Inventory was used. The statistical analysis includes: a descriptive analysis, the Student t test, and the analysis of the variance. RESULTS: Burnout scores were in the average range (41,9 +/- 7,5), with a more important grade in the decrease of personal accomplishment dimension (18,9 +/- 3,5). The 55,9% of the responders attributed to the conflict with the managing policy, their work unsatisfaction and the 58,8% planned to leave the job in the future. CONCLUSIONS: The importance of burnout and its three dimensions in the health professions and in the hospital setting is stated. It is enhanced that even though the burnout scores were in the average range more than half of the population studied were considering to leave their work in the future. PMID- 11003943 TI - [Dermatomyositis in childhood]. AB - AIM: The aim of this study is to review the presenting signs and symptoms, laboratory findings and therapeutic regimens of juvenile dermatomyositis in a tertiary hospital. METHODS: We reviewed retrospectively the available medical records of patients who met the clinicopathologic criteria of Bohan and Peter for definite juvenile dermatomyositis. They were followed between 1986 and july 1999 at the pediatric rheumatology section at our institution. RESULTS: The patient population included 3 male and 6 female patients. The mean age at diagnosis was 7 years. Clinical features demanding medical attention at the hospital were: muscle weakness and pain, with associated general symptoms in 4 cases; isolated muscle weakness in 2 cases; muscle weakness associated to general symptoms in 1 case; and monoarthritis in another case. One patient presented initially only with cutaneous rash. Three patients developed calcinosis. Serum CPK and LDH levels were elevated in 8 patients, aldolase in 7 and aminotransferases in 6. Autoantibodies were undetectable in all the patients. Electomyography demonstrated myopatic or mixed pattern in the 5 patients it was practiced. Muscle biopsy showed features of inflammatory myopathy in all the cases. Seven patients were treated exclusively with steroids and 2 patients with steroids, methotrexate and intravenous gammaglobuline. Actually, 6 patients are asymptomatic, 2 have mild muscle weakness, and 1 has died. CONCLUSIONS: The results of our review agree with other series reported. Juvenile dermatomyositis suspicious should be made upon muscle weakness and general symptoms. Treatment with steroids should be started promptly. PMID- 11003944 TI - [Proposed protocol for the study of cerebrovascular disease in childhood]. AB - AIM: The etiology of cerebrovascular disease in the paediatric population, remains unknown in up to 40% of the cases ("idiopathic"), but recent advances could improve this percentage. We devised a comprehensive study protocol for such investigation aimed at the identification of potentially modifiable risk factors for paediatric stroke. PATIENTS AND METHODS: From the 141 patients initially registered in our data base for stroke population (from January 1984 until December 1995), we invited all the patients with idiopathic cerebrovascular disease to complete the study protocol. New cases appeared from January 1996 until July 1999 were also included. RESULTS: A total of 68 cases were identified. We found an etiology in 38% and in 76% of the cases we found at least one risk factor for stroke. Mild hyperhomocysteinemia was the most frequent risk factor identified (36% of patients versus 5% of controls), one of them an infant with fatal haemorrhagic infarct with classic homocystinuria. 31% of the patients had thrombotic risk factors (protein S, protein C, antithrombine III deficiency, factor V Leiden, etc). 17.6% had unspecific febrile illness at the time of the cerebral infarction and 11.6% had minor head injuries before the stroke. CONCLUSIONS: The use of the protocol improves the identification of potentially modifiable risk factors for stroke in childhood and may serve as a practical guideline for clinicians. The stroke protocol is as important as management strategies for acute stroke or for recurrence prevention, currently under consideration in the adult population. PMID- 11003945 TI - [Lipid profile in children from Rivas-Vaciamadrid. Two-year follow-up]. AB - AIM: Evaluate the lipid profiles in children after two years of follow-up. METHODS: Longitudinal cohort study. A two years follow-up of a group of children since they were 6 years old. Blood analysis were carried in every children measuring total cholesterol, triglycerides, cLDL, cHDL, apoproteins A and B. It also included TD/cHDL, cLDL/cHDL, Apo B/A ratios and atherogenic index. RESULTS: 200 mg/dl, cLDL > 135 mg/dl and Apo B > 100 mg/dl in the follow-up was of 9.51, 5.18 and 8 for those children who had in the initial study these same values. The values of lipid profile and of the index studied improved in the two years of follow-up. This improvement has an statistical signification only in men. CONCLUSIONS: There is a significant correlation between the values of the lipid profile and the index measured in those children who were six years old in the beginning of the two years follow-up. PMID- 11003946 TI - [Blood pressure in childhood and adolescence. Its relationship with growth and maturation variables]. AB - AIM: To analyze the relationship between blood pressure in childhood and adolescence and different variables including gender, age, weight, height, body mass index and sexual maturation. METHODS: Cross-sectional study conducted in 1278 boys and girls aged 6 to 18 years old. Two blood pressure measurements were obtained. Mean value in the two measurements was obtained for each of the pressures: systolic (SBP), diastolic IV (DBPIV) and diastolic V (DBPV). Simple and multiple regression analysis were done to evaluate association between variables. RESULTS: 14 years 0.33; SBP boys > 14 years 0.34; DBPIV 0.25; DBPV 0.24) and height (SBP 6-13 years 0.33; SBP girls > 14 years 0.23; SBP boys > 14 years 0.29; DBPIV 0.25; DBPV 0.24) than for age (SBP 6-13 years 0.27; SBP girls > 14 years 0.17; SBP boys > 14 years 0.15; DBPIV 0.23; DBPV 0.22) for each of the blood pressures. Multiple regression analysis demonstrated a significant association between body mass index, height and SBP in the group of boys and girls aged 6-13 years old. No other significant associations were obtained. CONCLUSIONS: Blood pressure values in childhood and adolescence show a positive association with growth and maturation variables. It's of capital importance to considerate not only age, but also height, weight and body mass index when evaluating the blood pressure value of a child. PMID- 11003947 TI - [Aplasia cutis congenita in a newborn: etiopathogenic review and diagnostic approach]. AB - Aplasia cutis congenita is a rare condition characterized by the congenital absence of epidermis, dermis and, in some cases, subcutaneous tissues. It was first described by Cordon in 1767 and more than 500 cases have been reported since, with an estimate incidence of 3 in 10,000 births. The lesions may occur on any body surface although localised agenesis of the scalp is the most frequent pattern. In approximately 20% of cases underlying bone defects are also found. Aplasia cutis congenita occurs as an isolated defect or with other associated anomalies. There is no unifying theory for the pathogenesis and large scalp defects present a management dilemma. We report a newborn with a large scalp defect in the midline at the vertex without associated malformations. There was no significant family history. Skull and extremities radiographs, chromosome analysis, cerebral and abdominal sonography were normal. Two methods of treatment were used: a conservative approach consisting of daily antiseptic dressing to allow scalp epithelialization improved conditions for secondary surgery at 30 days of life, closing the defect with local rotational flaps. The postoperative course was uneventful and an excellent cosmetic result was achieved. PMID- 11003948 TI - [Delayed presentation of Bochdalek hernia associated with Hirschsprung disease]. PMID- 11003949 TI - [Congenital hepatic fibrosis and autosomal recessive polycystic kidney disease]. PMID- 11003950 TI - [Adrenocortical suppression after high doses of inhaled fluticasone]. PMID- 11003951 TI - [Reye-like syndrome as initial manifestation of mitochondrial disease]. PMID- 11003952 TI - [Magnetic resonance in a case of a newborn with pulmonary valve agenesis and deletion of chromosomal region 22q 11,2]. PMID- 11003953 TI - [Transhepatic cardiac catheterization in children. Report of one case]. PMID- 11003954 TI - [What is your diagnosis? Bloody diarrhea]. PMID- 11003955 TI - [What is your diagnosis? Causal image in an abdominal radiograph]. PMID- 11003956 TI - [Evidence-based paediatrics]. PMID- 11003957 TI - [Correlation between high plasma viral load and levels of TNF-alpha and cICAM-1 in HIV-1 infected children]. AB - AIM: To assess the relationship among plasma TNF-a and cICAM-1 levels and plasma viral load (VL) in HIV-infected children and to compare these values with those of healthy non-HIV infected children. PATIENTS AND METHODS: We studied 44 HIV infected children and 38 non-HIV-infected children. The VL was quantified using standard molecular assay. CD4 and CD8 lymphocyte subpopulations were determined by flow cytometry. TNF-a and cICAM-1 were quantified using commercially available specific enzyme-linked immunosorbent assay (ELISA). RESULTS: Levels of TNF-a and cICAM-1 were higher in HIV-infected children than in non-HIV infected children. HIV-infected children with VL > 50000 copies/ml had higher levels of TNF-a (12.83; 95% CI: 24.71 to 0.95 pg/ml) and cICAM-1 (248.94; 95% CI: 419.01 to 78.84 ng/ml) than HIV-infected children with VL < 50000 copies/ml. Interestingly, we found an increase of 6.57 pg/ml of TNF-a and 119.97 ng/ml of cICAM-1 levels for each log10 of VL. CONCLUSIONS: HIV-infected children had higher levels of TNF-a and cICAM-1 than healthy controls. Our data indicate a positive correlation among plasma TNF-a and cICAM-1 and VL levels. PMID- 11003958 TI - [Bone mineral density in juvenile-onset diabetes mellitus]. AB - OBJECTIVE: To determine bone mineral density (BMD) at axial and appendicular sites in patients with type 1 diabetes mellitus and evaluate its relationship with metabolic control and disease duration. PATIENTS AND METHODS: BMD was measured by dual-energy X-ray absorptiometry (Hologic QDR-1000) in the lumbar spine (L1-L4) and at the distal third forearm in 246 healthy non-diabetic children and adolescents (111 boys, 135 girls, aged 2.8-20.8 years) and in 45 diabetic patients (18 boys, 27 girls, aged 5.2-19.4 years). The results were expressed as the mean and standard deviation. The differences were tested by analysis of variance or Students t-test, as appropriate. The relationship between BMD and the remaining variables was studied by simple Pearsons coefficient and partial correlation coefficient. Significance was defined as p < 0.05. RESULTS: BMD Increased progressively from infancy to adulthood In both populations. Lumbar spine and forearm BMD were significantly lower in diabetic patients than in the healthy non-diabetic children, mainly during pubertal spurt. The greatest differences were found in males and in the trabecular bone. No relationship was found between metabolic control (mean glycosylated hemoglobin, insulin requirement) and duration of diabetes and the BMD in any region studied (p > 0.05). CONCLUSIONS: Pediatric patients with type 1 diabetes mellitus appear to constitute a population at risk of osteoporosis in adult-hood. Diagnosis of osteopenia diagnosis should be established according to densitometric criteria. In this study, metabolic control parameters and duration of diabetes did not enable predetermination of diabetic patients at risk of osteoporosis in adulthood. PMID- 11003959 TI - [Respiratory function in pediatric survivors of malignant neoplasms]. AB - AIM: To evaluate post-treatment pulmonary function in patients with malignant extrapulmonary neoplasia and its relationship with age, type of neoplasty and treatment received. METHODS: Cohort study of 95 pediatric patients after chemotherapy with or without surgery or extrapulmonary thoracic radiotherapy. The patients were in remission without treatment and able to undergo pulmonary function testing. Personal history and exposure to risk factors or toxic habits were evaluated. Clinical examination, chest radiographs, pulmonary gammography, basal and stress pulmometry, forced spirometry, whole body phletismography and carbonmonoxide transfer test were performed. RESULTS: Mean age at diagnosis was 5 +/- 3.3 years. Treatment duration was 2.4 +/- 1.3 years and time without treatment 4.3 +/- 3. 3 years. Thirty-six patients were reviewed two and a half years after the first control. Thirty-nine patients had acute lymphoblastic leukemia and 57 had solid tumors. Sixty percent showed functional restriction due to chest deformity after undergoing chest surgery. The association between functional restriction and radiotherapy (6 patients) did not produce poorer functional results. Forty-three percent showed initial change in TLCO. Nineteen percent showed basal hemoglobin saturation under 93% which in 16% fell after physical exercise. These alterations improved with time. However, restrictive change initially present in 11.5% persisted at the second evaluation. Thoracic surgery was the main cause of thoracic deformity and therefore of restrictive change. The children under 8 years old and those who received longer treatments tended to show the worst TLCO values. The patients with neuroblastoma showed greater restrictive change while the change in TLCO was more frequent in patients with Burkitt s lymphoma and in those treated with cyclophosphamide. CONCLUSIONS: Neither personal or family history of respiratory disease nor the presence of symptoms such as cough served to identify risk of functional change. Restrictive change in pulmonary function was greater in patients who had undergone thoracic surgery. Functional values were worse in patients with neuroblastoma. Pulmonary function should be followed up in pediatric survivors of malignant neoplasia in order to prevent restrictive alterations. PMID- 11003960 TI - [Food habits and dietary intake during the first year of life]. AB - OBJECTIVE: To determine feeding habits and dietetic ingestion du-ring the first year of life and to assess whether these are in accordance with dietetic recommendations. PATIENTS AND METHODS: A total of 171 healthy infants between 1 and 12 months old were studied. Two types of inquiry were carried out: 24-recall and weekly frequency of food. RESULTS: The percentage and duration of breastfeeding was low. Feeding with cows milk was more frequent than with breast milk from the age of 3 months. Thirty-eight per-cent of children had whole cows milk from the age of 6 months. Complementary feeding was introduced in accordance with international recommendations. The ingestion of energy and nutrients was also in accordance with the recommendations of the RDA during the first 6 months of life. In children aged 6-12 months, energy in-take was sufficient but diets were high in proteins and II-pIns and fairly high in carbohydrates. CONCLUSION: Ingestion of micronutrients and minerals are higher than is recommended except in the case of zinc. PMID- 11003961 TI - [Intellectual development in the second year of life in healthy breast-fed children compared with formula-fed children]. AB - BACKGROUND: The role of breast milk with regard to W3 long-chain polyunsaturated fatty acids and infant intellectual development remains controversial. PATIENTS AND METHODS: Thirty-nine children born at term and from homogeneous sociocultural status were enrolled in a blind-prospective trial. Children were divided in two non-randomized groups: a breast-fed group and a standard formula-fed group. Red blood cell phospholipid fatty acids were analyzed at 7 and 60 days of life. Cognitive development was evaluated at the end of the second year of life through Bailey s test. RESULTS: Concentrations of phosphatidylethanolamine and phosphatidylcholine docosahexaenoic acid were significantly lower in the formula fed group. No statistically significant differences between groups were found in cognitive function. Brain development index was significantly correlated with infant head circumference and educational status of the mother. CONCLUSIONS: Maternal nutrition provides higher red blood cell docosahexaenoic acid, but is not related to a higher developmental quotient at the age of 2 years. However, infant head circumference and maternal educational status were correlated with developmental cognition. PMID- 11003962 TI - [Admissions of patients with AIDS to pediatric intensive care units]. AB - OBJECTIVE: To describe the most significant clinical features of children with acquired immunodeficiency syndrome who required admission to a pediatric intensive care unit (PICU). METHODS: Retrospective study of 12 patients with AIDS who required 13 admissions, between January 1988 and December 1997. RESULTS: Mean age at admission was 15 months (1 month-6 years). Seven patients were under 1 year of age; four were diagnosed during their stay in the unit. The most common reason for admission was respiratory failure (six patients), followed by cardiac failure. Six patients needed mechanical ventilation (5 for respiratory failure). Two patients died during their stay, one of pneumonia due to Pneumocystis carinii infection and one of septic shock. CONCLUSIONS: One-third of patients was diagnosed with HIV Infection during their stay at the PICU. Opportunistic infection was the initial manifestation of the disease. Consequently, with this type of infection, clinical suspicion should be high. The survival rate of up to 84.6% of the admissions to our unit as well as the new, highly active antiretroviral therapy, generally make HIV-infected children suitable for treatment in intensive care units. PMID- 11003963 TI - [Children's unscheduled return visits to an emergency department]. AB - OBJECTIVE: To study the patients who make unscheduled return vi-sits in less than week to a pediatric emergency department (ED). PATIENTS AND METHOD: Retrospective review of 495 episodes in 233 children who were attended at our ED in November 1999. RESULTS: In November 1999, we registered 3667 episodes at our ED (495 corresponding to 233 children who made repeat visits, 13.4%). The most common chief complaints among children who revisited were fever and respiratory symptoms. The most common diagnoses were fever with-out apparent source, ORL infections, asthma and acute gastro-enteritis. The diagnoses made at the initial and fi-nal visits were the same in 131 cases (56.2%), a complication was detected in 18 (7.7%) and 84 (36.1%) were given a different diagnosis. The percentage of patients who received a symptomatic diagnosis was lower in the final visit. The admission rate of children who revisited was higher than that of children who visited our ED once (6.4% vs. 3.0%, p = 0.009), caused by the group of patients who received a different diagnosis when they revisited our ED. CONCLUSIONS: Patients who revisit a pediatric ED within a week are more likely to be admitted to the hospital, especially if the diagnosis they receive varies. PMID- 11003964 TI - [Abused children. Role of the pediatrician]. AB - Child maltreatment, in it's different forms (physical, negligence, Munchausen syndrome by proxy, sexual abuses.), represents an important morbidity cause, especially in the first years of the life. Nowadays, the battered child syndrome includes forms of abuse and different degrees of negligence (moderate, serious and light) in which the physical abuse could be absent. Determination of the forms of abuse, their diagnosis, intervention and prevention, corresponds to multidisciplinary teams, in which the pediatrician has a crucial role. PMID- 11003965 TI - [Treatment of very low birth weight infant: is it evidenced-based?]. AB - AIM: To determine what percentage of therapeutic interventions for very low birth weight infants undergoing neonatal intensive care is evidence based. METHODS: The management of 80 very low birth weight infants admitted to our neonatal unit during 1998 was retrospectively reviewed. For each clinical diagnosis e.g. respiratory distress syndrome, patent ductus arteriosus or chronic lung disease all interventions were recorder. Each intervention was then categorised according to the level of supporting evidence. Level I was supported by evidence from randomised controlled trials or meta-analysis of multiple trials. Level II included interventions backed by convincing non-experimental evidence where randomised controlled trials would be unnecessary or unethical. Level III were treatments in common use without substantial supporting evidence. These categorizations were made after extensive researching of Medline, The Cochrane Database and the Randomised Controlled Trial Register, detailed hand-searching of the literature as well as using local expertise and knowledge. RESULTS: 943 separate interventions were recorded in the charts of the 80 babies. Overall 91.3% were shown to be evidence-based of which 58.7% were level I, 32.6% were level II and only 8.7% were level III. CONCLUSIONS: 91.3% of interventions for very low birth weight infants in our neonatal intensive care unit were evidence based and only 8.7% had no substantial supporting evidence. Care of the very low birthweight infants is largely evidence-based. PMID- 11003966 TI - [Hereditary spherocytosis in neonates. Review of our caseload]. AB - AIM: Review of hereditary spherocytosis diagnosed in infants younger than two months and their follow up. PATIENTS AND METHODS: Retrospective study of 18 infants younger than two months diagnosed from 1973 to 1995. RESULTS: Diagnosis was established in the first week of life in 50% of the patients. Hereditary pattern was autosomic dominant in 94% of the cases. Anaemia was observed in all the patients and hyperbilirubinemia in only 44%, although the latter was the clinical presentation in patients diagnosed at younger age. Exchange transfusion was performed in 3 children (1 with the severe form and 2 with the typical form of the disease). During the first 6 months of age, 55% of infants presented hemolytic crises that required transfusion in 91% of them. Both periodicity of crises and transfusions decreased to 38 and 44% respectively after the first year. Splenectomy was performed in the 3 children with severe forms and in 6 with typical forms (mean age 8 years and 3 months). No cholecystectomy was required so far. CONCLUSIONS: The authors believe that neonatal spherocytosis does not implicate worse prognosis at follow up. Blood support is higher during the first year of life. Elective splenectomy depends on age and transfusional requirements. PMID- 11003967 TI - [Neurocutaneous melanosis]. AB - Neurocutaneous melanosis (NCM) is a rare congenital syndrome characterized by large or multiple congenital melanocytic nevi and excessive proliferation of melanotic cells in the leptomeninges. We report the case of a girl with a giant hairy nevus and numerous small nevi since birth. Within the first 2 years of life she developed clinical features of increased intracranial pressure and West s syndrome. At 2 years of age she presented a right facial palsy and myelopathy. Brain and spinal magnetic resonance imaging demonstrated meningeal infiltration. Diagnosis of NCM was established by a detailed cytologic analyses of the cerebrospinal fluid that revealed melanocytic cells. She received palliative treatment. The girl died 2 months after. Patients with large or multiple congenital melanocytic nevi should be carefully followed up with clinical examination and neuroimaging to detect NCM. At present there is no curative treatment. The association of NCM and West s syndrome has not been previously described. PMID- 11003968 TI - [Invasive disease caused by Streptococcus pyogenes. Report of a case with cutaneous and kidney metastasis]. AB - Few cases of invasive disease due to Streptococcus pyogenes in children have been described. Greater knowledge of this disease is needed due to its increasing incidence and seriousness. New serotypes have been discovered with greater virulence due to exotoxin production and to the development of resistance to phagocytosis. We present a case of invasive disease due to S. pyogenes with bacteremia and cutaneous and kidney metastasis, in a non-immunodeficient child. The case is of interest because kidney dissemination has not been described in the literature and because evolution was satisfactory with endovenous beta lactamica treatment during 15 days for only 48 hours. The importance of ruling out multi-organ metastatic foci, including use of urine cultures to rule out kidney involvement are highlighted. The epidemiological, pathogenic, clinical and diagnostic aspects of this entity, as reported in recent literature, are described. PMID- 11003969 TI - [Recurrent bowel intussusception and celiac disease]. AB - We report a 1-year-old girl with acute abdominal pain. Clinical examination revealed distended abdomen with increased intestinal peristalsis without other pathologic signs. Ultrasound of the abdomen showed a small bowel intussusception. Seven days later, after a new episode of sudden acute abdominal pain, ultrasound revealed a new bowel intussusception. Ig A and Ig G antigliadin-antibodies were elevated and intestinal biopsy revealed total villous atrophy. After being placed on a gluten-free diet the girl was free of colicky complaints and on repeated ultrasound there was no sign of intussusception. With recurrent intussusception, celiac disease should be considered as a cause. PMID- 11003970 TI - [Cervical mass. What is your diagnosis?]. PMID- 11003971 TI - In vitro effects of interleukin-10, prednisolone, and GM-CSF on the non-specific immune function of human polymorphonuclear leucocytes and monocytes. AB - A wide range of immune-modulating effects make IL-10 a potential therapeutic option in the treatment of numerous diseases pathophysiological based on a dysregulation of cytokine production. The background of this study was to investigate, whether the beneficial effects of a therapeutic immunosuppression with IL-10 may be countered by an increased risk for infections due to impaired effector cell functions of unspecific immunity. We demonstrated the in vitro effects of IL-10 on phagocytosis (P), intracellular killing (K), and chemotactic activity (C) by human neutrophils (PMN) and monocytes (MON) using Candida albicans as test strain and compared the results to the effects of prednisolone and GM-CSF. IL-10 reduced significantly the intracellular killing rate of PMN compared to untreated phagocytes (60 +/- 16% versus 68 +/- 13%, mean +/- SD, p = 0.0002). High dose IL-10 (100 ng/ml) had a stimulating effect on the percentage of phagocytizing MON (70.2 +/- 12.7% vs. 66.9 +/- 14.2%, p = 0.0436), without impairing intracellular killing. Prednisolone reduced significantly the Candida uptake by MON (57 +/- 18.1% vs. 66. 9 +/- 14.2%, p = 0.0019). In contrast to prednisolone, neither MON nor PMN chemotaxis was suppressed by IL-10. In conclusion, IL-10 had only marginal immunosuppressive effects on the unspecific immunity compared to prednisolone. PMID- 11003972 TI - Acute effects of 200 ppm 1,1,1-trichloroethane on the human EEG. AB - OBJECTIVES: Even low concentrations of organic solvents used at work may cause acute effects on the human central nervous system. We investigated the acute effects of 200 ppm 1,1,1-trichloroethane on the human EEG. METHODS: 12 healthy subjects were exposed for 4 hours to 200 ppm and to 20 ppm (control) in an exposure chamber in a cross-over design. EEG was recorded before (reference) and at the end of each exposure with eyes closed and open and during the Color Word Stress test. Spectral power was calculated by Fast Fourier transformation and related to reference values (per cent of baseline). Subjective symptoms and effects of blinding with 20 ppm 1, 1,1-trichloroethane were assessed by questionnaire. RESULTS: Blinding was not effective because of the strong smell of 1,1, 1-trichloroethane. The score for tiredness increased slightly during and after exposure to 200 ppm. In the closed eye condition, the median percentage of spectral power increased at all electrodes of the delta -band, significantly at temporo-occipital leads. In the theta-band, the percentage of the median spectral power was elevated at most of the electrodes but the parietal and some temporal ones. As to the alpha subset1-band, the percentage of the median spectral power was lower at the temporo-parieto-occipital electrodes, yielding significance at T subset4. In the alpha subset2-band, the percentage of the median spectral power was lower at all electrodes, significantly at T subset4 and T subset5. The percentage of the median spectral power of the temporo-parieto-occipital electrodes of the beta subset1 -band was lower during exposure to 200 ppm. There were no clear-cut changes in the beta subset2 -band, in the open eye condition and during the Color Word Stress test. CONCLUSION: The changes in EEG and the increased score for tiredness indicate a slight sedative effect of 200 ppm 1,1,1 trichloroethane. PMID- 11003973 TI - Effect of a fixed valerian-Hop extract combination (Ze 91019) on sleep polygraphy in patients with non-organic insomnia: a pilot study. AB - A pilot study with a fixed extract combination Ze 91019 of valerian and hop was conducted in 30 patients suffering from mild-moderate, non-organic insomnia. The diagnosis was confirmed by polysomnographic standard examinations. The patients were treated with 2 tablets in the evening. Each tablet contains 250 mg valerian extract and 60 mg hop extract. A polysomnographic re-examination after 2 weeks of treatment revealed declines in the sleep latency and the wake time. As a consequence the sleep efficiency increased. Sleep stage 1 (S1) was reduced and the slow wave sleep increased. In addition, the patients judged their being refreshed in the morning by assigning a rating of 1 to 6. They reported an improvement after 2 weeks of treatment. No adverse events were observed. Based on these findings a pivotal study can be designed. PMID- 11003974 TI - The therapeutic effect of theophylline in mild obstructive sleep Apnea/Hypopnea syndrome: results of repeated measurements with portable recording devices at home. AB - Theophylline has been recommended in the literature as a therapeutic option in mild OSAHS. In mild obstructive sleep apnea/hypopnea syndrome (OSAHS), night-to night variability of parameters of sleep-disordered breathing has been determined rarely, we therefore compared the results of serial measurements of sleep apnea/hypopnea parameters under the effects of placebo and theophylline. To this end, we measured the individual variability of the apnea-hypopnea index (AHI) for seven consecutive nights using a portable sleep apnea recording device, in 14 subjects (2 women, 12 men, mean age +/- [standard deviation] 50 +/- 8 years), treated with placebo or theophylline in a double blind, randomized crossover fashion, whose polysomnographically measured AHI was 13 +/- 5 /hour. Under theophylline treatment in comparison with placebo we observed a small but significant decrease in mean (of seven days) AHI at home (9.2 +/- 7.7 to 6.7 +/- 6.1 /hour), which was independent of body position. The night-to-night variability of AHI at home was high (9. 2 +/- 8.9 /hour; range: 0-31.4 /hour) and proved to be independent of body position and alcohol consumption. - In mild OSAHS, repeated measurements of sleep related breathing events should be performed. Theophylline showed a small but clinically insignificant potencial to reduce AHI. PMID- 11003975 TI - Intensified physiotherapy improves fitness to fly in cystic fibrosis patients. AB - OBJECTIVES: The life expectancy for patients with cystic fibrosis has risen constantly within the last 10 years to an average age over 20 years in western countries. However, most of these patients are partially respiratory insufficient. Frequently their physicians must decide whether it is acceptable to expose them to reduced air pressure on board of an aeroplane during long distance flights or how their fitness to fly can be improved prior to their annual vacation. We investigated the effect of intensified daily physiotherapy -carried out during a four weeks stay on Gran Canaria, Spain- on lung-function parameters, oxygen saturation and saliva neopterin as marker for ongoing inflammation in the respiratory tract. METHODS: 10 out of 12 patients (age 19 34 years, mean 28.6 years) were investigated two weeks prior and two weeks after their trip to the Canaries, Spain. Lung function on ground level and during flights to and from the island, oxygen saturation and saliva-flow adjusted neopterin in saliva were measured. RESULTS: Over the entire group vital capacity (VC) improved significantly from 2.89 l (range 1.37 3.95) to 3.36 l (range 1.23 5.24). Intrathoracic gas volume as a sign for pulmonary hyperinflation- declined slightly from 3.79 l (range 2.06 6.17) to 3.48 l (range 2. 18 5.85). The oxygen saturation increased from 89.5% (range 79 - 92%) during the flight to the Canaries to 94% (range 82 - 96%) during the return flight. Saliva neopterin dropped from 0.8 pmol/min. (range 0.2 6.8 pmol/min) to 0.6 ( range 0.1 2.6 pmol/min) pointing to diminished inflammation in the respiratory tract. CONCLUSION: Our results indicate that fitness to fly can be improved by intense physiotherapy as shown by improvement of vital capacity and oxygen saturation during flight. The influence of an additional climate change on the improvement of lung function in CF-patients needs further evaluation. PMID- 11003976 TI - Schistosomiasis and travel medicine. AB - The rise in off-track tourism and the spread of schistosomiasis in previously non endemic areas results in an increasing number of imported schistosomiasis cases. Detection of eggs in stool or urine and specific serology supports the diagnosis, however eosinophilia in early infection is low. In travel groups with a single index case all members of the group with similar risk behaviour should be examined since more than 50% of group members with similar risk behaviour are likely to be infected. Asymptomatic, atypical and late presentation of the disease might delay diagnosis. Praziquantel is not effective against immature worms, therefore treatment of returning travellers with single dose praziquantel is effective when started not earlier than 2 months after infection. A combination of praziquantel plus artemether is thought to be effective also in early stages of infection. Vaccination trials in endemic areas are in progress but a single vaccine for travellers covering all schistosoma species will not be available in the near future. Advising the traveller to avoid risk behaviour like wading, bathing and swimming in endemic fresh waters and about the risk of acquiring schistosomiasis through surface or spray water during boat tours will help to reduce imported schistosomiasis. PMID- 11003977 TI - Changes in cellular proliferation rate of lymphocytes after long-distance flights as a possible risk for patients with HIV-infection. AB - OBJECTIVES: Several studies showed that long-distance flights can influence cellular immunity. This might be due to a cortisol- and catecholamin- induced change in immunity with an impairment of T-lymphocyte dependent cellular immunity and an enhancement of B-lymphocyte dependent humoral immunity. Similar results can be found in patients with HIV. It is also known that progress of this disease and affection of T-helper-cells by the virus are induced by stimulation of the immune system, a phenomenon that also occurs during long distance flights. Therefore, a possible interaction between long-distance flights and the progression of HIV-infection should be discussed. METHODS: Cell cultures of 22 subjects after long-distance flights with and without rapid time zone shifts and of 16 patients with HIV (stage 2 3) were investigated. Mononuclear blood cells were stimulated with different lectins in culture and proliferation was measured by incorporation of bromodesoxyuridine. Moreover, all cultures were titrated with chromate concentrations between 0 to 700 ng/ml to measure the tolerance of the cells against chromate (VI) in vitro as a marker of the functional efficiency of the cellular part of the immune system. Maximal proliferation rate and tolerance against chromate were compared in both groups. RESULTS: After long distance flights tolerance against chromate decreased significantly during the first 24 h after flight. After 48 h levels were similar to those 1 week after flights. The decrease was similar to the results found in the stage 2 group of HIV-patients, but by far less to the decrease in stage 3 patients. Maximal proliferation rate dropped significantly during the second day after arrival compared to 1-week control values. CONCLUSION: Changes in the cellular immune system in healthy subjects after long-distance flights have been similar to the results of patients with stage 2 of HIV-infection. Mechanisms of changes in both groups are comparable in influencing T-cell-induced immunity. This could point to an additive effect on cellular immunity of HIV-patients by long distance-flights. Rosen neopterin concentrations and increases of apoptotic T-cells in both groups support this assumption. Therefore, further studies are urgently needed to investigate the interactions between HIV-infection and long-distance flights. PMID- 11003978 TI - Right ventricular systolic pressure load alters myocyte maturation in fetal sheep. AB - The effects of right ventricular (RV) systolic pressure (RVSP) load on fetal myocyte size and maturation were studied. Pulmonary artery (PA) pressure was increased by PA occlusion from mean 47.4 +/- 5.0 (+/-SD) to 71 +/- 13.6 mmHg (P < 0.0001) in eight RVSP-loaded near-term fetal sheep for 10 days. The maximal pressure generated by the RV with acute PA occlusion increased after RVSP load: 78 +/- 7 to 101 +/- 15 mmHg (P < 0.005). RVSP-load hearts were heavier (44.7 +/- 8.4 g) than five nonloaded hearts (31.8 +/- 0.2 g; P < 0.03); heart-to-body weight ratio (10.9 +/- 1.1 and 6.5 +/- 0.9 g/kg, respectively; P < 0.0001). RVSP RV myocytes were longer (101.3 +/- 10.2 microm) than nonloaded RV myocytes (88.2 +/- 8.1 microm; P < 0. 02) and were more often binucleated (82 +/- 13%) than nonloaded myocytes (63 +/- 7%; P < 0.02). RVSP-loaded myocytes had less myofibrillar volume than did nonloaded hearts (44.1 +/- 4.4% and 56. 1 +/- 2.6%; P < 0.002). We conclude that RV systolic load 1) leads to RV myocyte enlargement, 2) has minor effects on left ventricular myocyte size, and 3) stimulates maturation (increased RV myocyte binucleation). Myocyte volume data suggest that RV systolic loading stimulates both hyperplastic and hypertrophic growth. PMID- 11003979 TI - NF-kappaB mediates the protein loss induced by TNF-alpha in differentiated skeletal muscle myotubes. AB - Nuclear factor-kappaB (NF-kappaB) regulates the transcription of a variety of genes involved in immune responses, cell growth, and cell death. However, the role of NF-kappaB in muscle biology is poorly understood. We recently reported that tumor necrosis factor-alpha (TNF-alpha) rapidly activates NF-kappaB in differentiated skeletal muscle myotubes and that TNF-alpha acts directly on the muscle cell to induce protein degradation. In the present study, we ask whether NF-kappaB mediates the protein loss induced by TNF-alpha. We addressed this problem by creating stable, transdominant negative muscle cell lines. C2C12 myoblasts were transfected with viral plasmid constructs that induce overexpression of mutant I-kappaBalpha proteins that are insensitive to degradation via the ubiquitin-proteasome pathway. These mutant proteins selectively inhibit NF-kappaB activation. We found that differentiated myotubes transfected with the empty viral vector (controls) underwent a drop in total protein content and in fast-type myosin heavy-chain content during 72 h of exposure to TNF-alpha. In contrast, total protein and fast-type myosin heavy chain levels were unaltered by TNF-alpha in the transdominant negative cell lines. TNF-alpha did not induce apoptosis in any cell line, as assessed by DNA ladder and annexin V assays. These data indicate that NF-kappaB is an essential mediator of TNF-alpha-induced catabolism in differentiated muscle cells. PMID- 11003980 TI - Blunted arterial baroreflex causes "pathological" heart rate turbulence. AB - Sudden cardiac death is the leading cause of cardiovascular mortality in developed countries. Recently, two post-myocardial-infarction risk predictors were introduced that are superior to all other presently available indicators: turbulence onset (TO) and turbulence slope (TS). These parameters characterize the behavior of instantaneous heart rate after a ventricular premature beat, i.e., they describe the reestablishing of heart rate control after an acute perturbation. We propose that the dysfunction of an important cardiovascular control mechanism, the arterial baroreflex, is the mechanism behind these new potent markers. The hypothesis is tested by means of a physiological model involving the excitation generation in the heart, the hemodynamic situation in the aorta, and baroreceptor feedback mechanisms. The data show that a blunted baroreceptor response of the heart resembles patterns of heart rate turbulence that correspond to pathological values of TO and TS. The results of the model suggest that the recently established risk parameters TO and TS characterize baroreflex function, a known risk stratifier in patients. PMID- 11003981 TI - Hematological changes and athletic performance in horses in response to high altitude (3,800 m). AB - This study had two goals: 1) measure hematologic changes with high-altitude acclimatization in horses; and 2) assess the effect of 9 days at high altitude on subsequent athletic performance at low altitude. Six horses performed standardized exercise tests on a dirt track (before and during time at altitude) and treadmill (pre- and postaltitude exposure). Resting and immediate postexercise blood samples were measured for blood volume, lactate, red cell number, packed cell volume, and 2,3-diphosphoglycerate (DPG) concentrations at 225 m, over a 9-day period at 3,800 m, and shortly after returning to 225 m. Acclimatization produced increases in total red cell volume (38.2 +/- 2.4 to 48.1 +/- 2.9 ml/kg, P = 0.004) and DPG/hemoglobin concentrations (19.4 +/- 1.7 increased to 29.4 +/- 0. 4 micromol/g, P = 0.004). Two performance variables, heart rate recovery postexercise and lactate recovery, were faster after acclimatization. PMID- 11003982 TI - Testicular development in Siberian hamsters depends on frequency and pattern of melatonin signals. AB - We investigated the impact of frequency and pattern of melatonin signals on reproductive development in Siberian hamsters. Juvenile males gestated in short day lengths and housed in constant illumination to suppress melatonin secretion were infused with melatonin for 5 h either once or twice per day for 20 days. Melatonin infusions at either frequency produced equivalent increases in testes and body weights that exceeded those of animals infused with saline but were indistinguishable from those of hamsters transferred to long day lengths. The reproductive system appears to be maximally stimulated by a single short melatonin signal each day. Other animals kept from birth in a short photoperiod were treated 6 h after onset of darkness with the beta-adrenergic receptor antagonist DL-propranolol to shorten melatonin secretion on the night of injection but not on subsequent nights. This permitted interpolation of short nightly melatonin signals of 4-5 h duration against a background of long melatonin signals of 10-12 h duration on other nights. Treatment regimes that maintained a 1:1 ratio of short to long melatonin signals for 8 wk stimulated reproductive development; a 1:2 signal ratio, in each of three different patterns, was uniformly ineffective. The number of successive short melatonin signals had little influence on the interval across which successive melatonin signals were summated to influence photoperiodic traits. The neuroendocrine axis appears more responsive to short melatonin signal frequency than pattern for development of the summer phenotype. PMID- 11003983 TI - Panting in reindeer (Rangifer tarandus). AB - Two winter-insulated Norwegian reindeer (Rangifer tarandus tarandus) were exposed to air temperatures of 10, 20, 30, and 38 degrees C while standing at rest in a climatic chamber. The direction of airflow through nose and mouth, and the total and the nasal minute volumes, respectively, were determined during both closed- and open-mouth panting. The animals alternated between closed- and open-mouth panting, but the proportion of open-mouth panting increased with increasing heat load. The shifts from closed- to open-mouth panting were abrupt and always associated with a rise in respiratory frequency and respiratory minute volume. During open-mouth panting, the direction of airflow was bidirectional in both nose and mouth, but only 2.4 +/- (SD) 1.1% of the air was routed through the nose. Estimates suggest that the potential for selective brain cooling is markedly reduced during open-mouth panting in reindeer as a consequence of this airflow pattern. PMID- 11003984 TI - Red blood cell life span in the ovine fetus. AB - Red cell life span within the fetal circulation has not been reported, although erythrocyte life span has been studied in the adult and newborn. The present study quantified red cell life span in 12 chronically catheterized fetal sheep at 97-136 days gestation (term = 150 days) with the use of autologous red cells labeled with [(14)C]cyanate. Cyanate forms a permanent covalent bond with hemoglobin and acts as a permanent red cell label. In the fetuses, blood (14)C activity decreased in a curvilinear fashion with time and reached 50% of the initial activity at 16.4 +/- 1.6 (SE) days. In contrast, (14)C activity of autologous red cells in two adult ewes decreased linearly with time as expected, reached 50% of the initial (14)C activity in 59 days, and yielded life spans of 117 and 121 days. Computer modeling and parameter optimization taking into account growth and skewed life span distribution were used to analyze the (14)C disappearance curve in each fetus. The mean life span of all red cells in the fetal circulation was 63.6 +/- 5.8 days. Mean red cell life span increased linearly from 35 to 107 days as fetal age increased from 97 to 136 days (r = 0.83, P < 0.001). Life span of cells produced at the time of labeling was significantly greater than the mean life span. Fetal growth rate estimated from parameter optimization was 3.28 +/- 0.72%/day; this compared well with the rate of 3.40 +/- 0.14%/day calculated from fetal weights at autopsy. Mean corpuscular volume decreased as a function of gestational age, but the decrease was small compared with the large increase in red cell life span. We conclude the following: 1) red cell life span in the fetal circulation is short compared with the adult; 2) red cells in younger fetuses have shorter life spans than in near term fetuses; 3) the curvilinear disappearance of labeled red cells in the fetus appears to be due primarily to an expanding blood volume with fetal growth; and 4) red blood cell life span in a growing organism will be significantly underestimated unless the expansion of blood volume with growth is taken into account. PMID- 11003985 TI - Metabolic depression and enhanced O(2) affinity of mitochondria in hypoxic hypometabolism. AB - This study examined whether the steady-state hypometabolism seen in overwintering frogs (Rana temporaria) is reflected at the mitochondrial level either by a reduction in their resting (state 4) and active (state 3) respiration rates and/or by increases in O(2) affinity. We isolated mitochondria from the skeletal muscle of cold-submerged frogs at different stages during their hibernation in normoxic and hypoxic water. A modest metabolic depression at the whole animal level (normoxic submergence) was not associated with a reduction in mitochondrial state 4 and state 3 respiration rates. However, mitochondria isolated from frogs that were submerged for 1 mo manifested an increase in their O(2) affinity compared with controls and with animals submerged for 4 mo. Hypometabolism was more pronounced at the whole animal level during hypoxic submergence and was accompanied by 1) a reduction in mitochondrial state 4 and state 3 rates and 2) an increase in the O(2) affinity of mitochondria. These findings demonstrate that metabolic depression can be reflected at all levels of biological organization in hypoxia-tolerant animals. PMID- 11003986 TI - Substance P in the nucleus of the solitary tract augments bronchopulmonary C fiber reflex output. AB - Bronchopulmonary C fibers defend the lungs against injury from inhaled agents by a central nervous system reflex consisting of apnea, cough, bronchoconstriction, hypotension, and bradycardia. Glutamate is the putative neurotransmitter at the first central synapses in the nucleus of the solitary tract (NTS), but substance P, also released in the NTS, may modulate the transmission. To test the hypothesis that substance P in the NTS augments bronchopulmonary C fiber input and hence reflex output, we stimulated the C fibers with left atrial capsaicin (LA CAP) injections and compared the changes in phrenic nerve discharge, tracheal pressure (TP), arterial blood pressure (ABP), and heart rate (HR) in guinea pigs before and after substance P injections (200 microM, 25 nl) in the NTS. Substance P significantly augmented LA CAP-evoked increases in expiratory time by 10-fold and increases in TP and decreases in ABP and HR by threefold, effects prevented by neurokinin-1 (NK1) receptor antagonism. Thus substance P acting at NTS NK1 receptors can exaggerate bronchopulmonary C fiber reflex output. Because substance P synthesis in vagal airway C fibers may be enhanced in pathological conditions such as allergic asthma, the findings may help explain some of the associated respiratory symptoms including cough and bronchoconstriction. PMID- 11003987 TI - Intraportal glucose infusion and pancreatic islet blood flow in anesthetized rats. AB - The aim of the study was to evaluate whether a selective increase in portal vein blood glucose concentration can affect pancreatic islet blood flow. Anesthetized rats were infused (0.1 ml/min for 3 min) directly into the portal vein with saline, glucose, or 3-O-methylglucose. The infused dose of glucose (1 mg. kg body wt(-1). min(-1)) was chosen so that the systemic blood glucose concentration was unaffected. Intraportal infusion of D-glucose increased insulin release and islet blood flow; the osmotic control substance 3-O-methylglucose had no such effect. A bilateral vagotomy performed 20 min before the infusions potentiated the islet blood flow response and also induced an increase in whole pancreatic blood flow, whereas the insulin response was abolished. Administration of atropine to vagotomized animals did not change the blood flow responses to intraportal glucose infusions. When the vagotomy was combined with a denervation of the hepatic artery, there was no stimulation of islet blood flow or insulin release after intraportal glucose infusion. We conclude that a selective increase in portal vein blood glucose concentration may participate in the islet blood flow increase in response to hyperglycemia. This effect is probably mediated via periarterial nerves and not through the vagus nerve. Furthermore, this blood flow increase can be dissociated from changes in insulin release. PMID- 11003988 TI - GABAergic contribution to rat bladder hyperactivity after middle cerebral artery occlusion. AB - To evaluate the influences of gamma-aminobutyric acid (GABA) mechanisms on bladder hyperactivity after left middle cerebral artery occlusion, cystometric recordings were obtained from unanesthetized female rats. Intracerebroventricular administration of both muscimol (GABA(A) receptor agonist; 0.1-10 nmol) and baclofen (GABA(B) receptor agonist; 0.1-3 nmol) produced dose-dependent inhibitions of micturition with increases in bladder capacity (BC). The effects of high doses (1-10 nmol) were similar in sham-operated (SO) and cerebral infarcted (CI) rats. However, lower doses of muscimol (0.1 or 0.3 nmol) and baclofen (0.1 nmol) reduced BC in CI rats. After bicuculline (GABA(A) receptor antagonist; 1 or 3 nmol) administration, BC in both SO and CI rats first decreased and subsequently increased. An increase in urethral pressure was observed after administration of bicuculline (3 nmol) but not with either muscimol or baclofen. Infarct volumes in muscimol-, bicuculline-, or baclofen treated rats were not significantly different from those of vehicle-treated rats. These results suggest that GABAergic mechanisms inhibit the micturition reflex at the supraspinal level but that this can change as a result of CI. PMID- 11003989 TI - Gene expression in the rat supraoptic nucleus induced by chronic hyperosmolality versus hyposmolality. AB - Magnocellular neurons of the hypothalamo-neurohypophysial system play a fundamental role in the maintenance of body homeostasis by secreting vasopressin and oxytocin in response to systemic osmotic perturbations. During chronic hyperosmolality, vasopressin and oxytocin mRNA levels increase twofold, whereas, during chronic hyposmolality, these mRNA levels decrease to 10-20% of that of normoosmolar control animals. To determine what other genes respond to these osmotic perturbations, we have analyzed gene expression during chronic hyper- versus hyponatremia. Thirty-seven cDNA clones were isolated by differentially screening cDNA libraries that were generated from supraoptic nucleus tissue punches from hyper- or hyponatremic rats. Further analysis of 12 of these cDNAs by in situ hybridization histochemistry confirmed that they are osmotically regulated. These cDNAs represent a variety of functional classes and include cytochrome oxidase, tubulin, Na(+)-K(+)-ATPase, spectrin, PEP-19, calmodulin, GTPase, DnaJ-like, clathrin-associated, synaptic glycoprotein, regulator of GTPase stimulation, and gene for oligodendrocyte lineage-myelin basic proteins. This analysis therefore suggests that adaptation to chronic osmotic stress results in global changes in gene expression in the magnocellular neurons of the supraoptic nucleus. PMID- 11003990 TI - Mobilization of NK cells by exercise: downmodulation of adhesion molecules on NK cells by catecholamines. AB - The change of plasma catecholamine concentration correlates with the change of natural killer (NK) activity and NK cell number in peripheral blood mononuclear cells (PBMC) during and after moderate exercise. We studied the causal relation between exercise-induced catecholamine and expression of adhesion molecules on NK cells during and after exercise. The expression of CD44 and CD18 on CD3(-)CD56(+) NK cells was significantly reduced during exercise (P < 0.01). When PBMC were stimulated with 10(-8)M norepinephrine in vitro, the expression of these adhesion molecules on CD3(-)CD56(+) NK cells was downmodulated within 30 min. The binding capacity of NK cells to a CD44 ligand, hyaluronate, was reduced by the stimulation with norepinephrine (P < 0.01). The intravenous injection of norepinephrine in mice decreased the expression of CD44 and CD18 on CD3( )NK1.1(+) cells (P < 0.01) and increased the number of CD3(-)NK1.1(+) cells in PBMC (P < 0.01). These findings suggest that exercise-induced catecholamines modulate the expression of adhesion molecules on NK cells, resulting in the mobilization of NK cells into the circulation. PMID- 11003991 TI - Norepinephrine reuptake, baroreflex dynamics, and arterial pressure variability in rats. AB - This study examined the effect of norepinephrine reuptake blockade with desipramine (DMI) on the spontaneous variability of the simultaneously recorded arterial pressure (AP) and renal sympathetic nerve activity (SNA) in conscious rats. Acute DMI administration (2 mg/kg iv) depressed AP Mayer waves ( approximately 0.4 Hz) and increased low-frequency (<0.2 Hz) components of AP variability. DMI decreased renal SNA variability, especially due to the abolition of oscillations related to Mayer waves. To examine whether DMI-induced changes in AP and renal SNA variabilities could be explained by alterations in the dynamic characteristics of the baroreceptor reflex loop, the frequency responses of mean AP to aortic depressor nerve stimulation were studied in urethan-anesthetized rats. DMI accentuated the low-pass filter properties of the transfer function without significantly altering the fixed time delay. The frequency responses of iliac vascular conductance to stimulation of the lumbar sympathetic chain were studied in an additional group of anesthetized rats. DMI did not markedly alter the low-pass filter properties of the transfer function and slightly increased the fixed time delay. These results suggest that the DMI-induced decrease in the dynamic gain of the baroreceptor reflex is responsible for the decreased spontaneous renal SNA variability and the accompanying increased AP variability. The "slowing down" of baroreflex responses cannot be attributed to an effect of DMI at the vascular neuroeffector junction. PMID- 11003992 TI - Renal blood flow, early distal sodium, and plasma renin concentrations during osmotic diuresis. AB - Inconsistencies in previous reports regarding changes in early distal NaCl concentration (ED(NaCl)) and renin secretion during osmotic diuresis motivated our reinvestigation. After intravenous infusion of 10% mannitol, ED(NaCl) fell from 42.6 to 34.2 mM. Proximal tubular pressure increased by 12.6 mmHg. Urine flow increased 10-fold, and sodium excretion increased by 177%. Plasma renin concentration (PRC) increased by 58%. Renal blood flow and glomerular filtration rate decreased, however end-proximal flow remained unchanged. After a similar volume of hypotonic glucose (152 mM), ED(NaCl) increased by 3.6 mM, (P < 0.01) without changes in renal hemodynamics, urine flow, sodium excretion rate, or PRC. Infusion of 300 micromol NaCl in a smaller volume caused ED(NaCl) to increase by 6.4 mM without significant changes in PRC. Urine flow and sodium excretion increased significantly. There was a significant inverse relationship between superficial nephron ED(NaCl) and PRC. We conclude that ED(Na) decreases during osmotic diuresis, suggesting that the increase in PRC was mediated by the macula densa. The results suggest that the natriuresis during osmotic diuresis is a result of impaired sodium reabsorption in distal tubules and collecting ducts. PMID- 11003993 TI - Dissociation of leptin secretion and adiposity during prehibernatory fattening in little brown bats. AB - Hibernating animals deposit adipose tissue before hibernation to withstand long periods of reduced energy intake. Normally, adiposity is positively correlated with increased secretion from adipose tissue of the satiety hormone, leptin. During the prehibernatory phase of the little brown bat, Myotis lucifugus, body mass and adiposity increased to a maximum within 12 days. Leptin secretion from adipose tissue in vitro and plasma leptin, however, increased before the increase in adiposity, then significantly decreased when adiposity increased. Basal metabolic rate (BMR) decreased when plasma leptin was increasing. This was followed by an increase in nonshivering thermogenic capacity and brown adipose tissue mass. We conclude that in the early prehibernatory phase, BMR decreases despite increasing plasma leptin levels, suggesting a state of relative leptin resistance at that time. At later stages, adiposity increases as BMR continues to decrease, and plasma leptin becomes dissociated from adiposity. Thus, in M. lucifugus, hibernation may be achieved partly by removing the metabolic signal of leptin during the fattening period of prehibernation. PMID- 11003994 TI - Vascular reactivity and baroreflex function during hyperthermia in conscious rats. AB - The hemodynamic responses to vasoconstrictor agents are blunted during heating in anesthetized rats. It is unknown whether reflex neural responses to these agents are also altered during hyperthermia. Therefore, the purpose of this study was to determine the effect of hyperthermia on the hemodynamic and baroreflex-mediated sympathetic neural responses to vasoactive agents in conscious, unrestrained rats. The splanchnic sympathetic nerve activity (SpNA) and systemic and regional hemodynamic responses to injections of phenylephrine and sodium nitroprusside were measured during normothermia (37 degrees C) and hyperthermia (41.5 degrees C). The hemodynamic responses to phenylephrine and sodium nitroprusside were blunted with heating, whereas the SpNA responses to both agents were augmented or unchanged. At 41.5 degrees C, the baroreflex curves relating heart rate (HR) and SpNA to mean arterial blood pressure were shifted to the right. The operating range and gain of the blood pressure (BP)-HR reflex were significantly reduced during heating, whereas the operating range of the BP-SpNA reflex was augmented at 41.5 degrees C. These results indicate that heating alters the cardiovascular and sympathetic neural responses to vasoactive agents in vivo. Furthermore, the data suggest that heating differentially affects arterial baroreflex control of HR and SpNA, shifting both curves toward higher BP values but selectively attenuating baroreflex control of HR. PMID- 11003995 TI - Angiotensin II infusion to the midgestation ovine fetus: effects on the fetal kidney. AB - Renal and cardiovascular responses to an intravenous infusion of ANG II (1 microg/h) or saline for 3 days were examined in ovine fetuses at midgestation (75 85 days of gestation, term 150 days). ANG II caused an increase in fetal blood pressure (36 +/- 2 to 44 +/- 3 mmHg) and urine flow rate (8 +/- 2 to a maximum of 18 +/- 6 ml/h). Plasma renin concentrations decreased in ANG II-infused fetuses. Fetal fluids (amniotic and allantoic) did not differ in volume or composition between the groups when measured at postmortem. There was no difference in the expression levels of the mRNA for the angiotensin (AT(1) or AT(2)) receptors between the two groups when measured by an RNase protection assay. However, there was a significant decline in renin and AT(1) receptor gene expression when measured by a real-time polymerase chain reaction method. These results indicate that ANG II is diuretic and pressor when infused at midgestation. ANG II can feedback to decrease renin secretion by the fetal kidney, and this may occur by decreased renin gene expression. PMID- 11003996 TI - Impact of gender and endothelin on renal vasodilation and hyperfiltration induced by relaxin in conscious rats. AB - Chronic administration of the hormone relaxin elicits renal vasodilation that is dependent on nitric oxide (NO) in both conscious intact and ovariectomized female rats. Our first objective was to test whether the hormone, when administered to approximate serum concentrations found in midterm pregnant rats, induces renal vasodilation in males. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) increased significantly, on average, by 33 and 49% over baseline, respectively, after 5 days of recombinant human relaxin (rhRLX) administration to 12 conscious male rats by subcutaneous osmotic minipump. There were also significant decreases in hematocrit, plasma osmolality, and sodium concentration. Another objective was to determine whether endogenous endothelin (ET; via the endothelial ET(B) receptor) mediates the NO-dependent renal vasodilation produced by relaxin. rhRLX or vehicle was administered to conscious female rats (n = 9 and 8 rats, respectively). On the fifth day, baseline GFR and ERPF were both increased, on average, by 20-30% in the rats administered rhRLX (P < 0.05 vs. vehicle). Next, the specific ET(B)-receptor antagonist RES-701-1 was infused intravenously over 4 h in both groups of rats. In response to RES-701-1, there was a significant decline in both GFR and ERPF in the rats receiving rhRLX such that renal function converged in the two groups of animals. We conclude 1) relaxin induces marked changes in the renal circulation and in osmoregulation regardless of gender and 2) relaxin-induced renal vasodilation and hyperfiltration are mediated by endothelin through the endothelial ET(B) receptor subtype and NO. PMID- 11003997 TI - Impaired response of UCP family to cold exposure in diabetic (db/db) mice. AB - Impaired activity of the uncoupling protein (UCP) family has been proposed to promote obesity development. The present study examined differences in UCP responses to cold exposure between leptin-resistance obese (db/db) mice and their lean (C57Ksj) littermates. Basal UCP1 and UCP3 mRNA expression in brown adipose tissue was lower in obese mice compared with lean mice, but UCP2 expression in white adipose tissue (WAT) was higher. Basal skeletal muscle UCP3 did not change remarkably. The UCP family mRNAs, which were upregulated 12 and 24 h after cold exposure (4 degrees C), were returned to prior levels 12 h after rewarming exposure (21 degrees C) in lean mice. The accelerating effects of cold exposure on the UCP family were impaired in db/db obese mice. Together with these changes, WAT lipoprotein lipase mRNA was downregulated, and the concentration of serum free fatty acid was increased in response to cold exposure in the lean mice but not in db/db obese littermates. The impaired function of the UCP family and diminished lipolysis in response to cold exposure indicate that the reduced lipolytic activity may contribute to the inactivation of the UCP family in db/db obese mice. PMID- 11003998 TI - Conditioned suppression of contact sensitivity is independent of sympathetic splenic innervation. AB - The present study investigated the role of sympathetic innervation of the spleen in conditioned suppression of a contact hypersensitivity (CHS) reaction. Behavioral conditioning was achieved by pairing saccharin drinking solution (conditioned stimulus, CS) with injection of cyclosporin A (CsA, 20 mg/kg; unconditioned stimulus, UCS). Four days after sensitization of the animals by application of a 5% 2,4-dinitrochlorobenzene (DNCB) to abdominal skin, the animals were challenged by applying a 1% DNCB solution to the ear. The CHS response was monitored by measuring the degree of ear swelling. Saccharin re presentation reduced ear swelling to a magnitude that approached that achieved by CsA treatment. Histological examination demonstrated that the conditioned reduction of ear swelling was produced by a reduced leukocyte infiltration of the ear. Prior sympathetic denervation of the spleen did not alter the conditioned suppression of the CHS response. These data indicate that behavioral conditioning using CsA produces alterations of CHS that, unlike conditioned prolongation of heart allograft survival, are independent of sympathetically regulated conditioned alterations in the spleen. PMID- 11003999 TI - Circadian rhythm changes in core temperature over the menstrual cycle: method for noninvasive monitoring. AB - The purpose of this study was to determine whether core temperature (T(c)) telemetry could be used in ambulatory women to track changes in the circadian T(c) rhythm during different phases of the menstrual cycle and, more specifically, to detect impending ovulation. T(c) was measured in four women who ingested a series of disposable temperature sensors. Data were collected each minute for 2-7 days and analyzed in 36-h segments by automated cosinor analysis to determine the mesor (mean temperature), amplitude, period, acrophase (time of peak temperature), and predicted circadian minimum core temperature (T(c-min)) for each cycle. The T(c) mesor was higher (P < or = 0.001) in the luteal (L) phase (37.39 +/-0.13 degrees C) and lower in the preovulatory (P) phase (36.91 +/ 0.11 degrees C) compared with the follicular (F) phase (37.08 +/-0.13 degrees C). The predicted T(c-min) was also greater in L (37.06 +/- 0.14 degrees C) than in menses (M; 36.69 +/- 0.13 degrees C), F (36. 6 +/- 0.16 degrees C), and P (36.38 +/- 0.08 degrees C) (P < or = 0. 0001). During P, the predicted T(c-min) was significantly decreased compared with M and F (P < or = 0.0001). The amplitude of the T(c) rhythm was significantly reduced in L compared with all other phases (P < or = 0.005). Neither the period nor acrophase was affected by menstrual cycle phase in ambulatory subjects. The use of an ingestible temperature sensor in conjunction with fast and accurate cosinor analysis provides a noninvasive method to mark menstrual phases, including the critical preovulatory period. PMID- 11004000 TI - Treadmill running produces both positive and negative physiological adaptations in Sprague-Dawley rats. AB - Exercise training produces a vast array of physiological adaptations, ranging from changes in metabolism to muscle mitochondrial biogenesis. Researchers studying the physiological effects of exercise often use animal models that employ forced exercise regimens that include aversive motivation, which could activate the stress response. This study examined the effect of forced treadmill running (8 wk) on several physiological systems that are sensitive to training and stress. Forced treadmill running produced both positive and negative physiological adaptations. Indicative of positive training adaptations, exercised male Sprague-Dawley rats had a decrease in body weight gain and an increase in muscle citrate synthase activity compared with sedentary controls. In contrast, treadmill running also resulted in the potentially negative adaptations of adrenal hypertrophy, thymic involution, decreased serum corticosteroid binding globulin, elevated lymphocyte nitrite concentrations, suppressed lymphocyte proliferation, and suppressed antigen-specific IgM. Such alterations in neuroendocrine tissues and immune responses are commonly associated with chronic stress. Thus treadmill running produces both positive training adaptations and potentially negative adaptations that are indicative of chronic stress. Researchers employing forced activity need to be aware that this type of exercise procedure also produces physiological adaptations indicative of chronic stress and that these changes could potentially impact other measures of interest. PMID- 11004001 TI - Effect of gender on vestibular sympathoexcitation. AB - Studies have suggested that premenopausal women are more prone to orthostatic intolerance than men. Additionally, it has been postulated that the vestibulosympathetic reflex is important in regulating postural-related changes in sympathetic activity. The purpose of the present study was to determine whether men and women differ in their sympathetic and cardiovascular responses to stimulation of the otolith organs elicited by head-down rotation (HDR). Heart rate (HR), arterial pressure, calf blood flow (CBF), and leg muscle sympathetic nerve activity (MSNA) were measured during 3 min of HDR in the prone posture in 33 women and 30 men. With the exception of HR (71 +/- 2 and 63 +/- 1 beats/min for women and men, respectively; P < 0.01), all baseline variables were not different between genders. There were no gender differences in responses to HDR. MSNA increased 72 +/- 33 units (43%) in the men and 88 +/- 15 units (59%) in the women during HDR (P < 0.01). CBF decreased [-0.6 +/- 0.1 (15%) and -0.5 +/- 0.1 (19%) ml. min(-1). 100 ml(-1)] and calf vascular resistance increased [8 +/- 2 (21%) and 11 +/- 3 (25%) units during HDR for men and women, respectively (P < 0.01)]. Both in the men and women, HR increased 2 +/- 1 beats/min (P < 0.01). These results demonstrate that sympathetic activation during HDR in the prone posture is similar in men and women. Therefore, these findings suggest that the vestibulosympathetic reflex is not different between healthy men and women. PMID- 11004002 TI - Comparison of the effects of sucrose and fructose on insulin action and glucose tolerance. AB - The purpose of the present study was to determine whether fructose is the nutrient mediator of sucrose-induced insulin resistance and glucose intolerance. Toward this end, male rats were fed a purified starch diet (68% of total calories) for a 2-wk baseline period. After this, rats either remained on the starch (ST) diet or were switched to a sucrose (SU, 68% of total calories), fructose/glucose (F/G, 34/34% of total calories), or fructose/starch (F/ST, 34/34% of total calories) diet for 5 wk. Rats then underwent either an intravenous glucose tolerance test (n = 10/diet) or a euglycemic, hyperinsulinemic clamp (n = 8 or 9/diet). Incremental glucose and insulin areas under the curve in SU, F/G, and F/ST were on average 61 and 29% greater than ST, respectively, but not significantly different from one another. During clamps, glucose infusion rates (mg. kg(-1). min(-1)) required to maintain euglycemia were significantly lower (P < 0.05) in SU, F/G, and F/ST (13.4 +/- 0.9, 9. 5 +/- 1.7, 11.3 +/- 1.3, respectively) compared with ST (22.8 +/- 1. 1). Insulin suppression of glucose appearance (mg. kg(-1). min(-1)) was significantly lower (P < 0.05) in SU, F/G, and F/ST (5.6 +/- 0.5, 2.2 +/- 1.2, and 6.6 +/- 0.7, respectively) compared with ST (9.6 +/- 0.4). Insulin-stimulated glucose disappearance (mg. kg( 1). min(-1)) was significantly lower (P < 0.05) in SU, F/G, and F/ST (17. 9 +/- 0.6, 16.2 +/- 1.3, 15.3 +/- 1.8, respectively) compared with ST (24.7 +/- 1.2). These data suggest that fructose is the primary nutrient mediator of sucrose induced insulin resistance and glucose intolerance. PMID- 11004003 TI - Effect of neonatal hypoxia on the development of hepatic lipase in the rat. AB - Increases in plasma lipids occur during hypoxia in suckling but not in weaned rats and may result from altered hepatic enzyme activity. We exposed rats to 7 days of hypoxia from birth to 7 days of age (suckling) or from 28 to 35 days of age (weaned at day 21). Hypoxia led to an increase in hepatic lipid content in the suckling rat only. Hepatic lipase was decreased to approximately 45% of control in 7-day-old rats exposed to hypoxia but not in hypoxic 35-day-old rats. Hypoxic suckling rats also had a 50% reduction in lactate dehydrogenase activity, whereas transaminase activity and CYP1A and CYP3A protein content were not different between hypoxic and normoxic groups. Additional rats were studied 7 and 14 days after recovery from hypoxic exposure from birth to 7 days of age; hepatic lipase activity had recovered to 85% by 7 days and to 100% by 14 days in the rats previously exposed to hypoxia. Administration of dexamethasone to neonatal rats to simulate the hyperglucocorticoid state found in hypoxic 7-day-old rats led to a moderate decrease ( approximately 75% of control) in hepatic lipases. Developmentally, in the normoxic state, hepatic lipases increased rapidly after birth and reached levels more than twofold that of the newborn by 7 days of age. Hypoxia delays the maturation of hepatic lipases. We suggest that the decrease in hepatic lipase activity contributes to hyperlipemia in the hypoxic newborn rats. PMID- 11004004 TI - Amygdala-lesion obesity: what is the role of the various amygdaloid nuclei? AB - Anatomic descriptions of amygdaloid lesions resulting in hyperphagia and obesity in rats, cats, and dogs have been inconsistent and often contradictory, frequently resulting in failures to replicate. The present study attempted to reconcile these differences by examining common areas of overlap among differently placed lesions in female rats. Small bilateral lesions of the most posterodorsal aspects of the amygdala resulted in substantial weight gains (mean = 45.4 g/10 days). The smallest lesions caused damage limited to the posterodorsal medial amygdaloid nucleus and the bed nucleus of the stria terminalis and were directly in the area where axons are collecting to form the stria terminalis. Larger lesions that extensively damaged the central and/or anterodorsal medial amygdaloid nuclei sometimes resulted in excess weight gains, as did very large lesions of the basolateral nuclei, but substantial weight gains occurred only when the lesions extended (unilaterally or bilaterally) into the posterodorsal amygdala. Examination of previously published brain sections indicated that the hyperphagia and obesity that have been observed after widely differing lesion placements in cats and dogs were also the result of damage to a common area of overlap (i.e., the bed nucleus and/or stria terminalis). In rats, the critical area producing weight gain has extensive reciprocal relations with the medial hypothalamus. PMID- 11004005 TI - Differential stress responsivity in diet-induced obese and resistant rats. AB - The relationship between stress and obesity was assessed in male rats selectively bred to develop either diet-induced obesity (DIO) or diet resistance (DR) when fed a high-energy, 31% fat diet for 3 wk followed by 2 wk on a hyperphagic liquid diet (Ensure). One-half of the rats of each phenotype were subjected to moderate daily, unpredictable stress (cage changing, exposure to conspecific, swim, and immobilization stress, intraperitoneal saline injection) during the 5 wk. Both stressed and unstressed DIO rats were 26% heavier and ate 27% more than comparable DR rats at onset and had 48% lower basal morning plasma corticosterone levels. Stressed DR rats gained less weight and had significant elevations of basal morning corticosterone but reduced basal sympathetic activity (24-h urine norepinephrine) over 5 wk compared with their unstressed DR controls. Terminally, there was a 35% increase in the paraventricular nucleus corticotropin-releasing hormone mRNA expression. On the other hand, stressed DIO rats showed only a transient early increase in open-field activity and a terminal increase in basal corticosterone levels as the only effects of stress. Thus DIO rats are hyporesponsive to chronic stress compared with DR rats. This is in keeping with several other known differences in hypothalamopituitary and autonomic function in this model. PMID- 11004006 TI - Osteoblast tissue-nonspecific alkaline phosphatase antagonizes and regulates PC 1. AB - Tissue-nonspecific alkaline phosphatase (TNAP) is essential for bone matrix mineralization, but the central mechanism for TNAP action remains undefined. We observed that ATP-dependent (45)Ca precipitation was decreased in calvarial osteoblast matrix vesicle (MV) fractions from TNAP-/- mice, a model of infantile hypophosphatasia. Because TNAP hydrolyzes the mineralization inhibitor inorganic pyrophosphate (PP(i)), we assessed phosphodiesterase nucleotide pyrophosphatase (PDNP/NTPPPH) activity, which hydrolyzes ATP to generate PP(i). Plasma cell membrane glycoprotein-1 (PC-1), but not the isozyme B10 (also called PDNP3) colocalized with TNAP in osteoblast MV fractions and pericellular matrix. PC-1 but not B10 increased MV fraction PP(i) and inhibited (45)Ca precipitation by MVs. TNAP directly antagonized inhibition by PC-1 of MV-mediated (45)Ca precipitation. Furthermore, the PP(i) content of MV fractions was greater in cultured TNAP-/- than TNAP+/+ calvarial osteoblasts. Paradoxically, transfection with wild-type TNAP significantly increased osteoblast MV fraction NTPPPH. Specific activity of NTPPPH also was twofold greater in MV fractions of osteoblasts from TNAP+/+ mice relative to TNAP-/- mice. Thus TNAP attenuates PC 1/NTPPPH-induced PP(i) generation that would otherwise inhibit MV-mediated mineralization. TNAP also paradoxically regulates PC-1 expression and NTPPPH activity in osteoblasts. PMID- 11004007 TI - Circadian rhythms of body temperature and activity levels during 63 h of hypoxia in the rat. AB - The hypothermic response of rats to only brief ( approximately 2 h) hypoxia has been described previously. The present study analyzes the hypothermic response in rats, as well as level of activity (L(a)), to prolonged (63 h) hypoxia at rat thermoneutral temperature (29 degrees C). Mini Mitter transmitters were implanted in the abdomens of 10 adult Sprague-Dawley rats to continuously record body temperature (T(b)) and L(a). After habituation for 7 days to 29 degrees C and 12:12-h dark-light cycles, 48 h of baseline data were acquired from six control and four experimental rats. The mean T(b) for the group oscillated from a nocturnal peak of 38.4 +/- 0.18 degrees C (SD) to a diurnal nadir of 36.7 +/- 0.15 degrees C. Then the experimental group was switched to 10% O(2) in N(2). The immediate T(b) response, phase I, was a disappearance of circadian rhythm and a fall in T(b) to 36.3 +/- 0.52 degrees C. In phase II, T(b) increased to a peak of 38.7 +/- 0.64 degrees C. In phase III, T(b) gradually decreased. At reoxygenation at the end of the hypoxic period, phase IV, T(b) increased 1.1 +/- 0.25 degrees C. Before hypoxia, L(a) decreased 70% from its nocturnal peak to its diurnal nadir and was entrained with T(b). With hypoxia L(a) decreased in phase I to essential quiescence by phase II. L(a) had returned, but only to a low level in phase III, and was devoid of any circadian rhythm. L(a) resumed its circadian rhythm on reoxygenation. We conclude that 63 h of sustained hypoxia 1) completely disrupts the circadian rhythms of both T(b) and L(a) throughout the hypoxic exposure, 2) the hypoxia-induced changes in T(b) and L(a) are independent of each other and of the circadian clock, and 3) the T(b) response to hypoxia at thermoneutrality has several phases and includes both hypothermic and hyperthermic components. PMID- 11004008 TI - Contribution of central ANG II to acute stress-induced changes in baroreflex function in young rats. AB - The aim of the present investigation was to characterize the baroreflex in weaned 23- to 25-day-old rats when maternal influences were no longer present. The relationship between mean arterial pressure (MAP) and heart rate (HR) was determined during baroreceptor loading with phenylephrine and baroreceptor unloading with sodium nitroprusside in conscious rats, first in the freely moving state and subsequently during acute stress. In unstressed rats, the slope of the relationship between MAP and HR was greater during baroreceptor loading than baroreceptor unloading. Acute stress significantly attenuated the slope of the response to baroreceptor loading but increased the slope of the response to baroreceptor unloading. Pretreatment with intracerebroventricular or intravenous losartan, an AT(1) receptor antagonist, or intracerebroventricular alpha-helical corticotropin-releasing hormone (alpha-hCRH), a receptor antagonist, before the stress significantly reduced the stress-induced attenuation of slope during baroreceptor loading. Hence, young postweaning rats can alter baroreflex function during acute stress in a manner that would favor increases in MAP. Even at this young age, a central action of ANG II and CRH contributes to these stress-induced adaptations. PMID- 11004009 TI - Does angiotensin II have a significant tonic action on cardiovascular neurons in the rostral and caudal VLM? AB - The peptidic ANG II receptor antagonists [Sar(1),Ile(8)]ANG II (sarile) or [Sar(1),Thr(8)]ANG II (sarthran) are known to decrease arterial pressure and sympathetic activity when injected into the rostral part of the ventrolateral medulla (VLM). In anesthetized rabbits and rats, the profound depressor and sympathoinhibitory response after bilateral microinjections of sarile or sarthran into the rostral VLM was unchanged after prior selective blockade of angiotensin type 1 (AT(1)) and ANG-(1---7) receptors, although this abolished the effects of exogenous ANG II. Unlike the neuroinhibitory compounds muscimol or lignocaine, microinjections of sarile in the rostral VLM did not affect respiratory activity. Sarile or sarthran in the caudal VLM resulted in a large pressor and sympathoexcitatory response, which was also unaffected by prior blockade of AT(1) and ANG-(1---7) receptors. The results indicate that the peptidic ANG receptor antagonists profoundly inhibit the tonic activity of cardiovascular but not respiratory neurons in the VLM and that these effects are independent of ANG II or ANG-(1---7) receptors. PMID- 11004010 TI - Amiloride-sensitive signals and NaCl preference and appetite: a lick-rate analysis. AB - Rats prefer hypotonic and isotonic NaCl solutions to water in long-access drinking paradigms. To focus on the role of taste signals in NaCl preference, licking patterns of rats with 30-s exposure to NaCl solutions (0-0.5 M) were examined when they were either water deprived, sodium depleted, or not deprived (NaCl mixed in dilute sucrose). In all three conditions, rats displayed a preference for NaCl. The addition of 100 microM amiloride, a sodium channel blocker, to NaCl did not change rats' licking when they were sodium replete but dramatically reduced licking when they were deplete. Transection of the chorda tympani (CT) nerve, an afferent pathway for amiloride-sensitive Na(+) signals, had no effect on NaCl preference in nondeprived rats and only a modest effect on those that were Na(+) deplete. Amiloride was found to exert significant suppression of NaCl intake in Na(+)-depleted rats with transection of the CT, supporting the existence of other afferent pathways for transmission of amiloride sensitive Na(+) signalling. Together, these studies argue for the involvement of different neural signalling mechanisms in NaCl preference in the presence and absence of explicit Na(+) need. PMID- 11004011 TI - Detection of apical Na(+)/H(+) exchanger activity inhibition in proximal tubules induced by acute hypertension. AB - We previously showed that acute arterial hypertension induces an inhibition of fluid and NaCl reabsorption in proximal tubules of Sprague-Dawley rats, which is associated with a rapid reversible internalization of apical Na(+)/H(+) exchanger in brush border. To determine whether there is a corresponding inhibition of apical Na(+)/H(+) exchanger activity in proximal tubules to account for the reduced tubular reabsorption, an instrument capable of measuring intracellular pH (pH(i)) ratiometrically and repeatedly on the surface of kidney with high temporal resolution is required. We report the design and validation of such a fluorimetric system based on two ultraviolet nitrogen-pulsed lasers and a photomultiplier. pH(i) of proximal tubules in situ was measured with pH-sensitive fluorescence dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein at 5 Hz. Using the initial rate of change of pH(i) (dpH(i)/dt) during luminal Na(+) removal as an index of apical Na(+)/H(+) exchanger activity, the exchanger activity was found to be reduced by 52 +/- 11% (n = 14, P < 0.05) compared with the baseline after 20 min of induced acute hypertension. The inhibition of Na(+)/H(+) exchange activity was alleviated when the blood pressure was returned to prehypertensive level. These observations indicate that acute changes in arterial pressure can reversibly inhibit apical Na(+)/H(+) exchanger activity, which might contribute to pressure natriuresis in proximal tubule. PMID- 11004012 TI - Dual role of PKC in modulating pharmacomechanical coupling in fetal and adult cerebral arteries. AB - This study tested the hypothesis that protein kinase C (PKC) has dual regulation on norepinephrine (NE)-mediated inositol 1,4, 5-trisphosphate [Ins (1,4,5)P(3)] pathway and vasoconstriction in cerebral arteries from near-term fetal ( approximately 140 gestational days) and adult sheep. Basal PKC activity values (%membrane bound) in fetal and adult cerebral arteries were 38 +/- 4% and 32 +/- 4%, respectively. In vessels of both age groups, the PKC isoforms alpha, beta(I), beta(II), and delta were relatively abundant. In contrast, compared with the adult, cerebral arteries of the fetus had low levels of PKC-epsilon. In response to 10(-4) M phorbol 12,13-dibutyrate (PDBu; PKC agonist), PKC activity in both fetal and adult cerebral arteries increased 40-50%. After NE stimulation, PKC activation with PDBu exerted negative feedback on Ins(1,4,5)P(3) and intracellular Ca(2+) concentration ([Ca(2+)](i)) in arteries of both age groups. In turn, PKC inhibition with staurosporine resulted in augmented NE-induced Ins(1,4,5)P(3) and [Ca(2+)](i) responses in adult, but not fetal, cerebral arteries. In adult tissues, PKC stimulation by PDBu increased vascular tone, but not [Ca(2+)](i). In contrast, in the fetal artery, PKC stimulation was associated with an increase in both tone and [Ca(2+)](i). In the presence of zero extracellular [Ca(2+)], these PDBu-induced responses were absent in the fetal vessel, whereas they remained unchanged in the adult. We conclude that, although basal PKC activity was similar in fetal and adult cerebral arteries, PKC's role in NE-mediated pharmacomechanical coupling differed significantly in the two age groups. In both fetal and adult cerebral arteries, PKC modulation of NE-induced signal transduction responses would appear to play a significant role in the regulation of vascular tone. The mechanisms differ in the two age groups, however, and this probably relates, in part, to the relative lack of PKC-epsilon in fetal vessels. PMID- 11004013 TI - Baroreflex modulation of peripheral vasoconstriction during progressive hypothermia in anesthetized humans. AB - Mild hypothermia is a major concomitant of surgery under general anesthesia. We examined the hypothesis that baroreceptor loading/unloading modifies thermoregulatory peripheral vasoconstriction and, consequently, body core temperature in subjects undergoing lower abdominal surgery with general anesthesia. Thirty-six patients were divided into four groups: control group (C), applied positive end-expiratory pressure (PEEP; 10 cmH(2)O) group (P), applied leg-up position group (L), and a group of leg-up position patients with PEEP starting 90 min after induction of anesthesia (L + P). The esophageal temperature (T(es)) and the forearm-fingertip temperature gradient, as an index of peripheral vasoconstriction, were monitored for 3 h after induction of anesthesia. Mean arterial pressure and pulse pressure did not change during the study in any group. The change in right atrial transmural pressure from the baseline value was 0.3 +/- 0.1 mmHg in C, -3.0 +/- 0.5 mmHg in P, and 2.3 +/- 0.4 mmHg in L (P < 0.01). The change in T(es) at the end of the study was -1.7 +/- 0.1 (35.1 +/- 0.1) degrees C in C, -1.1 +/- 0.1 (35.7 +/- 0.1) degrees C in P, and -2.7 +/- 0.1 (34.1 +/- 0.1) degrees C in L, showing significant differences (P < 0.01). The T(es) threshold for thermal peripheral vasoconstriction was 35.6 +/- 0.1 degrees C in C, 36.2 +/- 0.2 degrees C in P, and 34.8 +/- 0.2 degrees C in L (P < 0.01). Excessive T(es) decrease in the leg-up-position operation was attenuated by applying PEEP (L + P group; P < 0.05). Our data indicate that baroreceptor loading augments and unloading prevents perioperative hypothermia in anesthetized and paralyzed subjects by reducing and increasing the body temperature threshold for peripheral vasoconstriction, respectively. PMID- 11004014 TI - Baroreflexes prevent neurally induced sodium retention in angiotensin hypertension. AB - Recent studies indicate that renal sympathetic nerve activity is chronically suppressed during ANG II hypertension. To determine whether cardiopulmonary reflexes and/or arterial baroreflexes mediate this chronic renal sympathoinhibition, experiments were conducted in conscious dogs subjected to unilateral renal denervation and surgical division of the urinary bladder into hemibladders to allow separate 24-h urine collection from denervated (Den) and innervated (Inn) kidneys. Dogs were studied 1) intact, 2) after thoracic vagal stripping to eliminate afferents from cardiopulmonary and aortic receptors [cardiopulmonary denervation (CPD)], and 3) after subsequent denervation of the carotid sinuses to achieve CPD plus complete sinoaortic denervation (CPD + SAD). After control measurements, ANG II was infused for 5 days at a rate of 5 ng. kg( 1). min(-1). In the intact state, 24-h control values for mean arterial pressure (MAP) and the ratio for urinary sodium excretion from Den and Inn kidneys (Den/Inn) were 98 +/- 4 mmHg and 1.04 +/- 0.04, respectively. ANG II caused sodium retention and a sustained increase in MAP of 30-35 mmHg. Throughout ANG II infusion, there was a greater rate of sodium excretion from Inn vs. Den kidneys (day 5 Den/Inn sodium = 0.51 +/- 0.05), indicating chronic suppression of renal sympathetic nerve activity. CPD and CPD + SAD had little or no influence on baseline values for either MAP or the Den/Inn sodium, nor did they alter the severity of ANG II hypertension. However, CPD totally abolished the fall in the Den/Inn sodium in response to ANG II. Furthermore, after CPD + SAD, there was a lower, rather than a higher, rate of sodium excretion from Inn vs. Den kidneys during ANG II infusion (day 5 Den/Inn sodium = 2.02 +/- 0.14). These data suggest that cardiac and/or arterial baroreflexes chronically inhibit renal sympathetic nerve activity during ANG II hypertension and that in the absence of these reflexes, ANG II has sustained renal sympathoexcitatory effects. PMID- 11004015 TI - Portal GLP-1 administration in rats augments the insulin response to glucose via neuronal mechanisms. AB - The incretin glucagon-like peptide-1 (GLP-1)-(7---36) amide is an important factor in prandial glucose homeostasis. Findings that GLP-1 is rapidly inactivated led to the hypothesis that the target of GLP-1 is close to the site of release. To investigate whether the target tissue is located in the hepatoportal system, we administered GLP-1 with glucose into the portal vein of rats and compared this with peripheral GLP-1 administration (jugular vein) and studied the effects of blockers of the nervous system. Portal GLP-1 augmented the insulin response to a portal glucose bolus by 81% (P < 0.01) and markedly improved the glucose disposal rate (P < 0.05). Peripheral administration of GLP-1 produced a similar augmentation of the insulin response (88%) and of the glucose disposal rate. However, only the effect of portal GLP-1 on insulin secretion was blocked by the ganglionic blocker chlorisondamine. The data suggest that prandial beta-cell stimulation by GLP-1 is evoked via a neural reflex triggered in the hepatoportal system. Because absorbed nutrients and GLP-1 first appear in the portal system, this mechanism may constitute a major pathway of GLP-1 action during meals. PMID- 11004016 TI - Synergy of L-arginine and growth hormone (GH)-releasing peptide-2 on GH release: influence of gender. AB - We test the hypotheses that 1) growth hormone (GH)-releasing peptide-2 (G) synergizes with L-arginine (A), a compound putatively achieving selective somatostatin withdrawal and 2) gender modulates this synergy on GH secretion. To these ends, 18 young healthy volunteers (9 men and 9 early follicular phase women) each received separate morning intravenous infusions of saline (S) or A (30 g over 30 min) or G (1 microg/kg) or both, in randomly assigned order. Blood was sampled at 10-min intervals for later chemiluminescence assay of serum GH concentrations. Analysis of covariance revealed that the preinjection (basal) serum GH concentrations significantly determined secretagogue responsiveness and that sex (P = 0.02) and stimulus type (P < 0.001) determined the slope of this relationship. Nested ANOVA applied to log-transformed measures of GH release showed that gender determines 1) basal rates of GH secretion, 2) the magnitude of the GH secretory response to A, 3) the rapidity of attaining the GH maximum, and 4) the magnitude or fold (but not absolute) elevation in GH secretion above preinjection basal, as driven by the combination of A and G. In contrast, the emergence of the G and A synergy is sex independent. We conclude that gender modulates key facets of basal and A/G-stimulated GH secretion in young adults. PMID- 11004017 TI - Synergy of L-arginine and GHRP-2 stimulation of growth hormone in men and women: modulation by exercise. AB - We investigated the ability of exercise, a multipathway, potent, physiological stimulus for GH release, to alter the synergistic interaction of L-arginine (A) and GH-related peptide (GHRP)-2 (G) observed at rest and the ability of gender to further modulate this putative interaction. Subjects (9 men and 9 early follicular phase women) completed 30 min of constant load aerobic exercise in combination with intravenous infusions of saline (S), A (30 g over 30 min), G (1 microg/kg bolus), or both (AG) in separate study sessions in randomly assigned order. Measures of GH release were logarithmically transformed for statistical analysis. Similar to rest, exercise maintained the rank order (AG > G > A > S) of effective stimulation of GH release for the key response measures in men or women, a gender disparity in the time to reach the maximal serum GH concentration, the calculated endogenous GH half-life, and the observed effect of preinfusion (basal) serum GH concentrations on determining secretagogue responsiveness. Exercise potentiated the individual stimulatory actions of A and G, while blunting the relative magnitude of the synergistic (supra-additive) interaction observed at rest. We infer from the present data that 1) exercise is likely to induce release of both GHRH and somatostatin, 2) L-arginine may facilitate the effect of exercise by limiting somatostatin release, 3) GHRP-2 could further enhance the stimulatory impact of exercise by opposing central actions of somatostatin and/or heightening endogenous GHRH release, and 4) gender strongly controls the relative but not absolute magnitude of A/G synergy both at rest and after exercise. PMID- 11004018 TI - Gastrointestinal motility during pregnancy: role of nitrergic component of NANC nerves. AB - This study evaluated whether increased release of nitric oxide (NO) from the nitrergic component of the nonadrenergic, noncholinergic (NANC) nerves may be partly responsible for the decrease in gastrointestinal motility observed during pregnancy. Segments of fundal strip, ileum, and colon were obtained from nonpregnant rats, rats in midpregnancy (days 9-11), and rats in late pregnancy (days 18-20). NANC activity was studied by assessing changes in tone after application of electric field stimulation (EFS). The role of NO was determined by observing the effects of EFS in the presence and absence of the NO synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) and the reversibility of the effects of L-NAME by L-arginine. The magnitude of change in cGMP levels in the tissues after application of EFS was also assessed. Our studies indicate that there was increased magnitude of relaxation of isolated strips of rat gastric fundus and rat colon, after application of EFS to tissues obtained only from animals in late pregnancy. These results paralleled the changes in cGMP levels in tissues. NOS activity in the gastric fundus was significantly increased in animals in late pregnancy compared with nonpregnant controls. Our studies suggest that the delay in gastric emptying and increase in colonic transit time observed in rats during pregnancy may be caused in part by increased activity of the nitrergic component of the NANC nerves innervating these organs. PMID- 11004019 TI - Cardiovascular and metabolic responses of hypertensive and normotensive rats to one week of cold exposure. AB - Challenges to energy homeostasis, such as cold exposure, can have consequences for both metabolic and cardiovascular functioning. We hypothesized that 1-wk cold exposure (4 degrees C) would produce concurrent increases in metabolic rate (VO(2); indirect calorimetry), heart rate (HR), and mean arterial blood pressure (MAP) measured by telemetry. In the initial hours of change in ambient temperature (T(a)), both spontaneously hypertensive rats (SHRs) and normotensive Sprague-Dawley rats showed rapid increases (in cold) or decreases (in rewarming) of VO(2), HR, and MAP, although the initial changes in MAP and HR were more exaggerated in SHRs. Throughout cold exposure, HR, VO(2), food intake, and locomotor activity remained elevated but MAP decreased in both strains, particularly in the SHR. During rewarming, all measures normalized quickly in both strains except MAP, which fell below baseline (hypotension) for the first few days. The results indicate that variations of T(a) produce rapid changes in a suite of cardiovascular and behavioral responses that have many similarities in hypertensive and normotensive strains of rats. The findings are consistent with the general concept that the cardiovascular responses to cold exposure in rats are closely related to and perhaps a secondary consequence of the mechanisms responsible for increasing heat production. PMID- 11004020 TI - Expression of members of the multidrug resistance protein family in human term placenta. AB - The placenta serves, in part, as a barrier to exclude noxious substances from the fetus. In humans, a single-layered syncytium of polarized trophoblast cells and the fetal capillary endothelium separate the maternal and fetal circulations. P glycoprotein is present in the syncytiotrophoblast throughout gestation, consistent with a protective role that limits exposure of the fetus to hydrophobic and cationic xenobiotics. We have examined whether members of the multidrug resistance protein (MRP) family are expressed in term placenta. After screening a placenta cDNA library, partial clones of MRP1, MRP2, and MRP3 were identified. Immunofluorescence and immunoblotting studies demonstrated that MRP2 was localized to the apical syncytiotrophoblast membrane. MRP1 and MRP3 were predominantly expressed in blood vessel endothelia with some evidence for expression in the apical syncytiotrophoblast. ATP-dependent transport of the anionic substrates dinitrophenyl-glutathione and estradiol-17-beta-glucuronide was also demonstrated in apical syncytiotrophoblast membranes. Given the cellular distribution of these transporters, we hypothesize that MRP isoforms serve to protect fetal blood from entry of organic anions and to promote the excretion of glutathione/glucuronide metabolites in the maternal circulation. PMID- 11004021 TI - Opioids affect acquisition of LiCl-induced conditioned taste aversion: involvement of OT and VP systems. AB - Aversive properties of lithium chloride (LiCl) are mediated via pathways comprising neurons of the nucleus of the solitary tract (NTS) and oxytocin (OT) and vasopressin (VP) cells in the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei. Because opioids act on brain regions that mediate effects of LiCl, we evaluated whether administration of opioids shortly before LiCl in rats influences 1) development of conditioned taste aversion (CTA) and 2) activation of NTS neurons and OT/VP cells. Neuronal activation was assessed by applying c-Fos immunohistochemical staining. Three opioids were used: morphine (MOR), a mu-agonist, butorphanol tartrate (BT), a mixed mu/kappa-agonist, and nociceptin/orphanin FQ (N/OFQ), which binds to an ORL1 receptor. BT and N/OFQ completely blocked acquisition of CTA. MOR alleviated but did not eliminate the aversive effects. Each of the opioids decreased LiCl-induced activation of NTS neurons as well as OT and VP cells in the PVN and SON. We conclude that opioids antagonize aversive properties of LiCl, presumably by suppressing activation of pathways that encompass OT and VP cells and NTS neurons. PMID- 11004022 TI - Angiotensin-converting enzyme inhibitor inhibits dehydration-enhanced fever induced by endotoxin in rats. AB - It has been reported that a host develops a marked fever under dehydrated conditions compared with normally hydrated conditions (11). The present study was carried out to investigate whether ANG II is involved in the enhancement seen in dehydrated rats of the fever induced by bacterial endotoxin. The results showed that intravenous injection of bacterial endotoxin produced a fever in dehydrated rats (rats deprived of water for 24 h) that was significantly greater than that seen in normally hydrated rats. In contrast, dehydration had no effect on the fever induced by intravenous interleukin-1beta (IL-1beta). Under dehydrated conditions, the enhanced endotoxin-induced fever was significantly inhibited by the angiotensin-converting enzyme inhibitor lisinopril, but the IL-1beta fever was not. These results suggest that the dehydration-induced enhancement of endotoxin fever is due, at least in part, to the action of ANG II, which elicits an increased production of pyrogenic cytokines such as IL-1. PMID- 11004023 TI - Don't try this at home! PMID- 11004024 TI - A century of regional analgesia in obstetrics. PMID- 11004025 TI - The lack of benefit of tracheal extubation in the operating room after coronary artery bypass surgery. AB - Although early tracheal extubation in cardiac anesthesia is safe and cost beneficial, questions still remain regarding how early after cardiac surgery patients should be tracheally extubated (TE). Our objective was to determine the effects on resource use if patients scheduled for coronary artery bypass grafting have TE in the operating room (OR). We studied 100 consecutive patients undergoing elective coronary artery bypass grafting, requiring extracorporeal circulation, and those eligible for a fast-track pathway. At the end of the procedure, the patients were evaluated for TE in the OR if they were hemodynamically stable, were without significant bleeding, and fulfilled clinical and blood gas analysis variables. Patients who did not meet the requirements had TE in the intensive care unit (ICU). Fifty patients had TE in the OR and 50 patients in the ICU. Time in the OR after skin closure, ICU length of stay, and postoperative length of stay were similar between the groups. Four patients (8%) in the OR group were tracheally reintubated secondary to respiratory depression (P = 0.11). Three patients (6%) in the OR group had postoperative myocardial infarction, and one postoperative myocardial infarction (2%) occurred in the ICU group (P = 0.61). All four patients recovered satisfactorily. The incidences of other complications were similar between groups. PMID- 11004026 TI - A comparison of superficial versus combined (superficial and deep) cervical plexus block for carotid endarterectomy: a prospective, randomized study. AB - Carotid endarterectomy may be performed by using cervical plexus blockade with local anesthetic supplementation by the surgeon during surgery. Most practitioners use either a superficial cervical plexus block or a combined (superficial and deep) block, but it is unclear which offers the best operative conditions or greatest patient satisfaction. We compared the two techniques in patients undergoing carotid endarterectomy. Forty patients undergoing carotid endarterectomy were randomized to receive either a superficial or a combined cervical plexus block. Bupivacaine 0.375% to a total dose of 1.4 mg/kg was used. The main outcome measure was the amount of supplemental lidocaine 1% used by the surgeon. Subsidiary outcome measures were postoperative pain score, sedative and analgesic requirements before and during surgery, and postoperative analgesic requirements. Median supplemental lidocaine requirements were 100 mg (range 30 180 mg) in the superficial block group and 115 mg (range 30-250 mg) in the combined block group. These differences were not statistically significant (Mann Whitney U-test). There was no significant difference in the number of patients needing postoperative analgesia between the groups (11 of 20 in the deep block group versus 8 of 20 in the superficial block group) in the 24 h after surgery. The median time to first analgesia in the superficial block group was 150 min, more than in the combined block group (median time 45 min) but this difference, although large, was not statistically significant (Mann-Whitney U-test). We found no significant differences between the anesthetic techniques studied. All patients reported satisfaction with the techniques. PMID- 11004027 TI - Left stellate ganglion block has only small effects on left ventricular function in awake dogs before and after induction of heart failure. AB - Left stellate ganglion block (LSGB) results in acute sympathetic denervation of the left ventricular (LV) posterobasal wall. We investigated the effects of LSGB in chronically instrumented awake dogs before and after the induction of pacing induced congestive heart failure. Twelve dogs were instrumented for measurement of global hemodynamics [LV pressure (LVP)], its first derivative (dP/dt), cardiac output (CO), and regional myocardial function (systolic posterobasal segment length shortening, mean velocity [SLmv]). Before the induction of heart failure (n = 12), LSGB did not affect CO [3.2+/-1.4 (control, mean +/- SD) vs. 3.3+/-1.6 L/min (LSGB, P = 0.45)] and SLmv (11.1+/-4.0 vs. 10.8+/-4.0 mm/s, P = 0.16), but slightly reduced LVP (130+/-12 vs. 125+/-14 mm Hg, P = 0.04), dP/dt(max) (3614+/ 755 vs. 3259+/-644 mm Hg/s, P = 0.003) and dP/dt(min) (-3153+/-663 vs. -2970+/ 725 mm Hg/s, P = 0.03). During heart failure (n = 8), global hemodynamics [CO (2.8+/-1.2 vs. 2.7+/-1.2 L/min, P = 0.04), LVP (119+/-6 vs. 112+/-9 mm Hg, P = 0.01), dP/dt(max) (1945+/-520 vs. 1824+/-554 mm Hg/s, P = 0.03) and dP/dt(min) ( 2402+/-678 vs. -2243+/-683 mm Hg/s, P = 0.04)], as well as regional myocardial function, were significantly different after LSGB [SLmv] (8.0+/-3.8 vs. 6.9+/-3.4 mm/s, P = 0.02)]. In conclusion, even during heart failure, the hemodynamic changes after LSGB are small, confirming its broad margin of safety. PMID- 11004028 TI - The effects of clonidine on human digital vasculature. AB - Large concentrations of alpha(2) agonists cause vasoconstriction. However, the threshold of the vasoconstrictive effect in humans is not known. We studied seven volunteers to determine the lower limit of the vasoconstrictive effect of clonidine. Subjects were studied while they were awake, and they were anesthetized with propofol/alfentanil/N(2)O. Arterial blood pressure was continuously monitored via radial arterial catheter and vasoconstriction via finger volume plethysmography measuring infrared light transmitted through a fingertip (LTF). Clonidine was administered, targeting plasma clonidine concentrations of 0.3, 0.45, 0.68, 1.0, 1.5, and 2.25 ng/mL. The maximum change from preclonidine values for systolic blood pressure (SBP) and LTF was analyzed by using repeated measures analysis of variance. In awake subjects, clonidine (2.25 ng/mL) decreased LTF by 14%+/-13% and SBP from 141+/-7 to 110+/-15 mm Hg (P<0.0001). In contrast, clonidine (2.25 ng/mL) increased LTF in anesthetized subjects by 21%+/-16% and SBP from 91+/-7 to 106+/-19 mm Hg (P<0.0001). We conclude that the same dose of clonidine that decreased blood pressure and caused vasodilation in awake subjects had the opposite effect in anesthetized subjects with reduced sympathetic tone, increasing blood pressure and causing vasoconstriction in human digital vasculature. Our findings suggest that the lower threshold for clonidine-induced vasoconstriction in human digital vasculature is 1.0 ng/mL. PMID- 11004029 TI - Optison (FS069) disrupts the blood-brain barrier in rats. AB - Optison is a new echocardiographic contrast agent, designed for IV injection, that is very useful in delineating cardiac structures during ultrasound examination. Because Optison could be a valuable adjunct in the diagnosis and evaluation of congenital heart disease, this study was undertaken to assess its effects on the blood-brain barrier when introduced directly in the cerebral circulation, as might occur with some congenital lesions. In this study, Sprague Dawley rats were anesthetized, and Optison, at various dosages, was injected into the carotid artery. After this, Evans blue dye, a marker for blood-brain barrier disruption, was injected at different time intervals. Gross and histologic examination of the animals' brains revealed disruption of the blood-brain barrier that appeared to be Optison-dosage-dependent. Although the mechanism for this disruption is unclear, it may be related to the use of octofluoropropane gas used in the Optison as a contrast medium. Further studies are necessary to determine the pathologic consequences of Optison's effects on the blood-brain barrier. PMID- 11004030 TI - Randomized safety studies of intravenous perflubron emulsion. I. Effects on coagulation function in healthy volunteers. AB - Previous perfluorocarbon (PFC) emulsions have been associated with transient adverse events (i.e., platelet activation, decreased platelet count, febrile responses, changes in hemodynamic function). The Phase I studies described in this report were parallel, randomized, double-blinded, placebo-controlled studies conducted in 48 healthy volunteers (n = 24 per study) with perflubron emulsion (Oxygent; Alliance Pharmaceutical Corp., San Diego, CA). Because of the decreased platelet counts observed with previous PFC emulsions and the intended use of perflubron emulsion in surgical patients, these studies assessed postdosing coagulation responses and hemostasis. PFC pharmacokinetic variables were also evaluated. The primary endpoint for examination of coagulation effects was prospectively defined as bleeding time. Subjects received either saline (3 mL/kg) control, or perflubron emulsion at 1.2 g PFC/kg or 1.8 g PFC/kg, and were evaluated for a 14-day period. No postinfusion changes in bleeding time or differences in ex vivo agonist-induced platelet aggregation were observed. A 17% reduction in platelet count was observed 3 days after dosing in the 1.8-g PFC/kg group; levels recovered to baseline by Day 7. The intravascular half-life of perflubron for the first 24 h was dose dependent: 9.4+/-2.2 h and 6.1+/-1.9 h in the 1.8- and 1.2-g PFC/kg groups, respectively. Results indicate that this perflubron emulsion did not affect coagulation function in healthy volunteers. PMID- 11004031 TI - Randomized safety studies of intravenous perflubron emulsion. II. Effects on immune function in healthy volunteers. AB - Particle size distribution is a major determinant of particle clearance by the mononuclear phagocytic system and the potential for concomitant activation of resident macrophages. To test the safety of a second-generation perflubron-based emulsion (60% perfluorocarbon [PFC] wt/vol; Oxygent [Alliance Pharmaceutical Corp., San Diego, CA]) with a small mean particle size, two parallel, randomized, double-blinded, placebo-controlled studies were conducted in 48 healthy volunteers (n = 24 per study). The study described herein focuses on safety concerning immune function. The primary endpoint was defined prospectively as delayed hypersensitivity skin test responses with lymphocyte proliferative responses to mitogenic stimulation providing a secondary measure for changes in cell-mediated immunity. Subjects received either perflubron emulsion IV (1.2 g PFC/kg or 1.8 g PFC/kg) or saline (3 mL/kg) control. Perflubron emulsion had no effect on delayed hypersensitivity skin reactions, lymphocyte proliferative potential, circulating immunoglobulins, complement activation, or plasma levels of the inflammatory cytokines, tumor necrosis factor-alpha, interleukin-1 alpha, and interleukin-1 beta. Perflubron emulsion was generally well tolerated, although there was a dose-dependent increase in minor flu-like symptoms in the perflubron treatment groups at 24 h after dosing. Increased serum levels of interleukin-6 were observed in those subjects exhibiting febrile responses. The clinical safety profile of perflubron emulsion supports its continued investigation as a temporary oxygen carrier in surgical patients to reduce exposure to allogeneic blood transfusion. PMID- 11004032 TI - The effect of hydroxyethyl starch 200 kD on platelet function. AB - We evaluated the effects of hydroxyethyl starch with a molecular weight of 200 kD (HES 200 kD) on platelets to gain insight into the potential mechanisms involved in the anticoagulant effects of HES 200 kD. Blood was obtained before and after an IV infusion (10 mL/kg) of either saline (n = 15) or HES 200 kD (n = 15) in otherwise healthy patients scheduled for minor elective surgery. Flow cytometry was used to assess the expression of glycoprotein (GP) IIb-IIIa, GP Ib, and P selectin on agonist-activated platelets. Overall platelet function was evaluated by assessing thromboelastographic maximum amplitude (MA) in celite-activated blood and platelet function analyzer-closure times by using collagen/adenosine diphosphate cartridges. Saline infusion had no effects on platelet variables, whereas HES 200 kD reduced GP IIb-IIIa expression and MA and prolonged platelet function analyzer-closure times, without affecting the expression of P-selectin and GP Ib. In vitro experiments extended these observations by a concentration related inhibiting effect of HES 200 kD on GP IIb-IIIa expression. This study demonstrates that cellular abnormalities with decreased availability of platelet GP IIb-IIIa are involved in the anticoagulant effects of HES 200 kD. PMID- 11004033 TI - The effect of clonidine premedication on hemodynamic responses to microlaryngoscopy and rigid bronchoscopy. AB - The usual hemodynamic response to laryngoscopy and bronchoscopy is an increase in heart rate and arterial blood pressure. Previous work has reported that 10%-18% of the patients develop ischemic ST segment changes during the procedure. Therefore, we performed a prospective, randomized, double-blinded study in 36 patients scheduled for elective microlaryngeal and bronchoscopic surgical procedures to evaluate the effects of 300-microg oral clonidine premedication (n = 18) or placebo (n = 18) on the hemodynamic alterations and the incidence of perioperative myocardial ischemic episodes. Myocardial ischemia was assessed by using continuous electrocardiographic monitoring, beginning 30 min before, and lasting until 24 h after the operation. During the procedure, patients receiving placebo exhibited a significant increase (mean +/- SD) in arterial blood pressure (the systolic increasing from 137+/-11 to 166+/-17 mm Hg, the diastolic increasing from 80+/-11 to 97+/-14 mm Hg) and heart rate (increasing from 79+/-15 to 97+/-12 bpm) compared with the baseline and with the clonidine group. A dose of 300-microg clonidine blunted the hemodynamic response to endoscopy. Ventricular arrhythmias were more frequent in patients who were not premedicated with clonidine. Two patients in the control group, but none in the clonidine group, had evidence of myocardial ischemia. These data should encourage routine premedication with clonidine in patients undergoing microlaryngoscopic and bronchoscopic procedures. PMID- 11004034 TI - Smaller-than-expected sevoflurane concentrations using the Sevotec 5 vaporizer at low fill states and high fresh gas flows. AB - Smaller-than-expected concentrations of sevoflurane were delivered by Sevotec 5 vaporizers (Datex-Ohmeda, Madison, WI) at low-fill states, 8% dialed concentrations, and high fresh gas flows. Clinically, this would lead to prolonged induction times. The risk of complications from prolonged inhaled induction could be increased when using a vaporizer with these characteristics. PMID- 11004035 TI - Specific genetic diseases at risk for sedation/anesthesia complications. AB - We reviewed of a number of genetic diseases known or at risk for sedation or anesthesia complications. Some of these conditions are relatively common (e.g., Down's syndrome) whereas others are rare or present with multiple congenital anomalies that have an impact on health care delivery. We listed complications, recommended presedation evaluations, and included checklist items to assist the health care provider administering sedation and anesthesia. A better recognition and awareness of risk factors associated with specific genetic diseases should lessen the likelihood of complications during these procedures. IMPLICATIONS: This article provides a brief description of potential problematic genetic disorders and associated complications that may manifest during sedation or anesthesia. Recommendations for presedation evaluation and checklist items are given that may impact on the delivery of care for these patients. PMID- 11004036 TI - Transient neurologic symptoms after epidural analgesia in a five-year-old child. AB - Transient neurologic symptoms occurred in a five-year-old child after discontinuation of epidural analgesia with bupivacaine and morphine, which had been established for two days after thoracotomy. The symptoms resolved within 12 hours with acetaminophen. PMID- 11004037 TI - Spinal anesthesia for gastrostomy in an infant with nemaline myopathy. AB - IMPLICATIONS: We report a case of spinal anesthesia in an infant with nemaline myopathy. Spinal anesthesia was administered to preempt the potential problems of difficult tracheal intubation and prolonged mechanical ventilation secondary to muscle weakness after the use of IV anesthetics in this patient. PMID- 11004038 TI - A comparison of spinal, epidural, and general anesthesia for outpatient knee arthroscopy. AB - We compared general, epidural, and spinal anesthesia for outpatient knee arthroscopy (excluding anterior cruciate ligament repairs). Forty-eight patients (ASA physical status I-III) were randomized to receive either propofol-nitrous oxide general anesthesia with a laryngeal mask airway with anesthetic depth titrated to a bispectral index level of 40-60, 15-20 mL of 3% 2-chloroprocaine epidural, or 75 mg of subarachnoid procaine with 20 microg fentanyl. All patients were premedicated with <0.035 mg/kg midazolam and <1 microg/kg fentanyl and received intraarticular bupivacaine and 15-30 mg of IV ketorolac during the procedure. Recovery times, operating room turnover times, and patient satisfaction were recorded by an observer using an objective scale for recovery assessment and a verbal rating scale for satisfaction. Statistical analysis was performed with analysis of variance and chi(2). Postanesthesia care unit discharge times for the general and epidural groups were similar (general = 104+/ 31 min, epidural = 92+/-18 min), whereas the spinal group had a longer recovery time (146+/-52 min) (P = 0.0003). Patient satisfaction was equally good in all three groups (P = 0.34). Room turnover times did not differ among groups (P = 0.16). There were no anesthetic failures or serious adverse events in any group. Pruritus was more frequent in the spinal group (7 of 16 required treatment) than in the general or epidural groups (no pruritus) (P<0.001). We conclude that epidural anesthesia with 2-chloroprocaine provides comparable recovery and discharge times to general anesthesia provided with propofol and nitrous oxide. Spinal anesthesia with procaine and fentanyl is an effective alternative and is associated with a longer discharge time and increased side effects. IMPLICATIONS: For outpatient knee arthroscopy, anesthesia can be provided adequately with regional or general anesthesia. Epidural and general anesthesia provide equal recovery times and patient satisfaction, whereas spinal anesthesia may prolong recovery and have increased side effects. The choice of anesthesia may depend primarily on the patient's interest in being alert or asleep during the procedure. PMID- 11004039 TI - A comparison of minidose lidocaine-fentanyl and conventional-dose lidocaine spinal anesthesia. AB - The syndrome of transient neurologic symptoms (TNS) after spinal lidocaine has been presumed to be a manifestation of local anesthetic neurotoxicity. Although TNS is not associated with either lidocaine concentration or dose, its incidence has never been examined with very small doses of spinal lidocaine. One hundred ten adult ASA physical status I and II patients presenting for arthroscopic surgery of the knee were randomly assigned to receive spinal anesthesia with either 1% hypobaric lidocaine 50 mg (Group L50) or 1% hypobaric lidocaine 20 mg + 25 microg fentanyl (Group L20/F25). Hemodynamic data, block height and regression, and time to first micturition and discharge were recorded. Follow-up phone calls were made by a blinded researcher at 48-72 h using a standardized questionnaire. Both groups had a median peak cephalad block level of T10. Lidocaine 50 mg was associated with a greater decrease in systolic blood pressure and a greater need for ephedrine. Time until block regression to the S2 dermatome (80 vs. 110 min) and outpatient time to void (130 vs 162 min) and discharge (145 vs. 180 min) were faster in the L20/F25 group. Complaints of TNS were found in 32.7% of the patients in the L50 group and in 3.6% of the patients in the L20/F25 group. We conclude that spinal anesthesia with lidocaine 20 mg + fentanyl 25 microg provided adequate anesthesia with greater hemodynamic stability and faster recovery than spinal anesthesia with lidocaine 50 mg. The incidence of TNS after spinal lidocaine 20 mg + fentanyl 25 microg was significantly less than that after spinal lidocaine 50 mg. PMID- 11004040 TI - Bisulfite-containing propofol: is it a cost-effective alternative to Diprivan for induction of anesthesia? AB - Propofol (Diprivan(TM); AstraZeneca, Wilmington, DE) is a commonly used drug for the induction of general anesthesia in the ambulatory setting. With the availability of a new bisulfite-containing generic formulation of propofol, questions have arisen regarding its cost effectiveness and safety compared with Diprivan(TM). Two hundred healthy outpatients were randomly assigned, according to a double-blinded protocol, to receive either Diprivan(TM) or bisulfite containing propofol 1.5 mg/kg IV as part of a standardized induction sequence. Maintenance of anesthesia consisted of either desflurane (4%-8% end-tidal) or sevoflurane (1%-2% end-tidal) in combination with a remifentanil infusion (0.125 microg x kg(-1) x min(-1) IV). Patient assessments included pain on injection, induction time, hemodynamic and bispectral electroencephalographic changes during induction, emergence time, and incidence of postoperative nausea and vomiting. The two propofol groups were comparable demographically, and the induction times and bispectral index values during the induction were also similar. However, the bisulfite-containing formulation was associated with less severe pain on injection (5% vs 11%), with fewer patients recalling pain on injection after surgery (38% vs. 51%, P<0.05). None of the patients manifested allergic-type reactions after the induction of anesthesia. The acquisition cost (average wholesale price in US dollars) of a 20-mL ampoule of Diprivan(TM) was $15 compared with $13 for the bisulfite-containing propofol formulation. Therefore, we concluded that the bisulfite-containing formulation of propofol is a cost effective alternative to Diprivan(TM) for the induction of outpatient anesthesia. IMPLICATIONS: Bisulfite-containing propofol and Diprivan(TM) (AstraZeneca, Wilmington, DE) were similar with respect to their induction characteristics; however, the generic formulation was associated with a smaller incidence of injection pain. Assuming that the drug costs are similar, these data suggest that the bisulfite-containing formulation of propofol is a cost-effective alternative to Diprivan(TM). PMID- 11004041 TI - Recovery profiles and costs of anesthesia for outpatient unilateral inguinal herniorrhaphy. AB - The use of an ilioinguinal-hypogastric nerve block (IHNB) as part of a monitored anesthesia care (MAC) technique has been associated with a rapid recovery profile for outpatients undergoing inguinal herniorrhaphy procedures. This study was designed to compare the cost-effectiveness of an IHNB-MAC technique with standardized general and spinal anesthetics techniques for inguinal herniorrhaphy in the ambulatory setting. We randomly assigned 81 consenting outpatients to receive IHNB-MAC, general anesthesia, or spinal anesthesia. We evaluated recovery times, 24-h postoperative side effects and associated incremental costs. Compared with general and spinal anesthesia, patients receiving IHNB-MAC had the shortest time-to-home readiness (133+/-68 min vs. 171+/-40 and 280+/-83 min), lowest pain score at discharge (15+/-14 mm vs. 39+/-28 and 34+/-32 mm), and highest satisfaction at 24-h follow-up (75% vs. 36% and 64%). The total anesthetic costs were also the least in the IHNB-MAC group ($132.73+/-33.80 vs. $172.67+/-29.82 and $164.97+/-31.03). We concluded that IHNB-MAC is the most cost-effective anesthetic technique for outpatients undergoing unilateral inguinal herniorrhaphy with respect to speed of recovery, patient comfort, and associated incremental costs. PMID- 11004042 TI - Percutaneous tracheostomy: ciaglia blue rhino versus the basic ciaglia technique of percutaneous dilational tracheostomy. AB - Percutaneous dilational tracheostomy (PDT), according to Ciaglia's technique described in 1985, has become the most popular technique for percutaneous tracheostomy and is demonstrably as safe as surgical tracheostomy. In 1999, an extensively modified technique of PDT was introduced, the Ciaglia Blue Rhino (CBR; Cook Critical Care, Bloomington, IL), that consists of one-step dilation by means of a curved dilator with hydrophilic coating. To compare CBR with the basic technique of PDT, we performed a prospective, randomized trial in 50 critically ill adults. Twenty-five of these patients had PDT, and 25 had CBR. Average operating times were <3 min for CBR (range: 50-360 s) and <7 min for PDT (range: 4-20 min; P<0.0001). Tracheostomy was successfully completed in all patients. When CBR was performed, 11 minor, nonlife-threatening complications were noted: nine fractures of tracheal cartilage and two short periods of intraoperative oxygen desaturation. During PDT, seven complications occurred, of which three were potentially life-threatening: two injuries to the posterior tracheal wall, one pneumothorax, two tracheal cartilage fractures (P< 0.05 vs CBR), one case of bleeding, and one short episode of intraoperative oxygen desaturation. Regardless of whether PDT or CBR was performed, oxygenation was not significantly affected, and there was no infection of the tracheostoma. Based on our data, we conclude that new CBR is more practicable than PDT. No life-threatening complications occurred during CBR. IMPLICATIONS: To assess practicability and safety of the Ciaglia Blue Rhino (Cook Critical Care, Bloomington, IL)-an extensively modified technique of percutaneous dilatational tracheostomy-50 critically ill adults on long-term ventilation underwent either new Ciaglia Blue Rhino or percutaneous dilatational tracheostomy in a prospective, randomized clinical trial. PMID- 11004043 TI - The good, the bad, and the ugly: should we completely banish human albumin from our intensive care units? AB - IMPLICATIONS: Human albumin is still widely used in critically ill patients for volume replacement therapy or for correcting hypoproteinemia. Most meta-analyses on the value of albumin administration are over 15 yr old and raise more questions than they answer. With the help of a MEDLINE analysis, we examined more recent studies in humans using albumin. Most of these studies have recommended a very cautious use of albumin in critically ill patients. PMID- 11004044 TI - Endotoxin augments cerebral hyperemic response to halothane by inducing nitric oxide synthase and cyclooxygenase. AB - We examined the cerebral hyperemic response to halothane after treatment with bacterial lipopolysaccharide (LPS). To determine the involvement of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2), we tested whether the effect of LPS on halothane-induced hyperemia was altered by pretreatment with the selective iNOS inhibitor, aminoguanidine (100 mg/kg), COX-2 inhibitor, NS-398 (5 mg/kg), or enzyme expression inhibitor, dexamethasone (4 mg/kg). Further, we examined whether the administration of a nitric oxide donor, diethylamine NONOate, would change the cerebral hyperemic response of halothane. Sprague Dawley rats were anesthetized with 0.5 minimum alveolar anesthetic concentration of halothane and artificially ventilated. Regional cerebrocortical blood flow (rCBF) was assessed by laser-Doppler flowmetry. LPS (1 mg/kg) was administered intracerebroventricularly; artificial cerebrospinal fluid was used in controls. Four hours after LPS infusion, iNOS and COX-2 messenger ribonucleic acid (mRNA) levels (reverse transcription-polymerase chain reaction) and enzyme activities (arginine-citrulline conversion and prostaglandin E(2) enzyme immunoassay) were significantly increased. LPS enhanced halothane-induced 3.9 and 1.6-fold increases in rCBF at 1.0 and 1.5 minimum alveolar concentration, respectively. Co treatment with NS-398 attenuated, but aminoguanidine or dexamethasone abolished the effect of LPS on halothane-induced rCBF increase. Diethylamine NONOate mimicked the enhanced rCBF response to halothane. These results suggest that LPS augmented halothane-induced cerebrocortical hyperemia by induction of iNOS and COX-2. PMID- 11004045 TI - A comparative study between a calcium channel blocker (Nicardipine) and a combined alpha-beta-blocker (Labetalol) for the control of emergence hypertension during craniotomy for tumor surgery. AB - We compared the efficacy of the combination of enalaprilat/labetalol with that of enalaprilat/nicardipine to prevent emergence postcraniotomy hypertension. A prospective, randomized open labeled clinical trial was designed to compare the incidence of breakthrough hypertension (systolic blood pressure [SBP] > 140 mm Hg) and adverse effects (hypotension, tachycardia, and bradycardia) between the two drug combinations. Secondarily, the effects of the drugs on SBP, mean blood pressure, and diastolic blood pressure were evaluated over the course of the study. Forty-two patients received enalaprilat 1.25 mg IV at dural closure followed by either multidose nicardipine 2 mg IV or labetalol 5 mg IV to maintain the SBP below 140 mm Hg. SBP was similarly controlled in both groups. There was a marginally smaller incidence of failures and adverse effects with labetalol. Blood pressure profiles were similar for both groups. PMID- 11004046 TI - Arm pain as an unusual presentation of postdural puncture intracranial hypotension. AB - IMPLICATIONS: We report a case of a patient experiencing severe arm pain after dural puncture. This complication has not been reported previously. The patient was successfully treated with an epidural patch. PMID- 11004047 TI - Crack cocaine, myocardial infarction, and troponin I levels at the time of cesarean delivery. AB - IMPLICATIONS: During the peripartum period, cocaine-abusing women are highly susceptible to myocardial infarction. This report describes a case of myocardial infarction diagnosed by increased troponin I levels in a pregnant patient with a history of recent crack cocaine use and severe preeclampsia. PMID- 11004048 TI - Postdural puncture headache: a randomized comparison of five spinal needles in obstetric patients. AB - This prospective, blinded, randomized study compares the incidence of postdural puncture headache (PDPH) and the epidural blood patch (EBP) rate for five spinal needles when used in obstetric patients. One thousand two women undergoing elective cesarean delivery under spinal anesthesia were recruited. We used two cutting needles: 26-gauge Atraucan and 25-gauge Quincke, and three pencil-point needles: 24-gauge Gertie Marx (GM), 24-gauge Sprotte, and 25-gauge Whitacre. The needle for each weekday was chosen randomly. Cutting needles were inserted parallel to the dural fibers. The incidences of PDPH were, respectively, 5%, 8.7%, 4%, 2.8%, and 3.1% for Atraucan, Quincke, GM, Sprotte, and Whitacre needles (P = 0.04, chi(2) analysis), and the corresponding EBP rates in those with PDPH were 55%, 66%, 12.5%, 0%, and 0% (P = 0.000). The Quincke needle had a more frequent PDPH rate than the Sprotte or the Whitacre needle (P = 0.02) and a more frequent EBP rate than the GM, Sprotte, or the Whitacre needle (P = 0.01). The Atraucan needle had a more frequent EBP rate than the Sprotte or Whitacre needle (P = 0.05). Neither the PDPH rate nor the EBP rates differed among the pencil point needles. The cost of EBP must be taken into consideration when choosing a spinal needle. We conclude that pencil-point spinal needles should be used for subarachnoid anesthesia in obstetric patients. PMID- 11004049 TI - Drug use inefficiency: a hidden source of wasted health care dollars. AB - A potential area for departmental savings is to minimize inefficient use of pharmaceuticals. We recorded drug waste data for multiple drugs for a fiscal year and surveyed providers' knowledge of departmental drug waste. Six large-cost or large-volume use drugs were chosen for study: thiopental, succinylcholine, rocuronium, atracurium, midazolam, and propofol. Amounts administered to patients were collected for one year by using a computerized anesthesia record keeper. Total drug distributed was the number of vials restocked by pharmacy for the year. An efficiency index, the percent administered to patients, was calculated for each drug. Drug administration to 25,481 patients was analyzed. Drug use efficiency indices were: atracurium 29%; thiopental, 31%; succinylcholine, 33%; propofol, 49%; midazolam, 53%; rocuronium, 61%. The total cost of unadministered study drugs was $165,667, 26% of the expenditure for all drugs. Most dollars wasted were for propofol, $80,863, and thiopental, $32,839. The reason most cited for drug waste was the disposal of full, or partially full, syringes. Drug wastage represents a significant portion of the entire anesthesia drug budget. Waste reduction strategies should allow a portion of the "avoidable" waste to be reduced. IMPLICATIONS: Unadministered drug amounts were measured for six study drugs over one fiscal year and found to be significant; the cost of unadministered drugs totaled $165,667. The reason most cited for waste was disposal of full, or partially full, syringes. PMID- 11004050 TI - A strategy to decide whether to move the last case of the day in an operating room to another empty operating room to decrease overtime labor costs. AB - We examined how to program an operating room (OR) information system to assist the OR manager in deciding whether to move the last case of the day in one OR to another OR that is empty to decrease overtime labor costs. We first developed a statistical strategy to predict whether moving the case would decrease overtime labor costs for first shift nurses and anesthesia providers. The strategy was based on using historical case duration data stored in a surgical services information system. Second, we estimated the incremental overtime labor costs achieved if our strategy was used for moving cases versus movement of cases by an OR manager who knew in advance exactly how long each case would last. We found that if our strategy was used to decide whether to move cases, then depending on parameter values, only 2.0 to 4.3 more min of overtime would be required per case than if the OR manager had perfect retrospective knowledge of case durations. The use of other information technologies to assist in the decision of whether to move a case, such as real-time patient tracking information systems, closed circuit cameras, or graphical airport-style displays can, on average, reduce overtime by no more than only 2 to 4 min per case that can be moved. IMPLICATIONS: The use of other information technologies to assist in the decision of whether to move a case, such as real-time patient tracking information systems, closed-circuit cameras, or graphical airport-style displays, can, on average, reduce overtime by no more than only 2 to 4 min per case that can be moved. PMID- 11004051 TI - A magnetic resonance imaging study of modifications to the infraclavicular brachial plexus block. AB - A previously described infraclavicular brachial plexus block may be modified by using a more lateral needle insertion point, while the patient abducts the arm 45 degrees or 90 degrees. In performing the modified block on patients abducting 45 degrees, we often had problems finding the cords of the brachial plexus. Therefore, we designed an anatomic study to describe the ability of the recommended needle direction to consistently reach the cords. Additionally, we assessed the risk of penetrating the pleura by the needle. Magnetic resonance images were obtained in 10 volunteers. From these images, a virtual reality model of each volunteer was created, allowing precise positioning of a simulated needle according to the modified block, without exposing the volunteers to actual needle placement. In both arm positions, the recommended needle angle of 45 degrees to the skin was too shallow to reach a defined target on the cords. Comparing the two arm positions, target precision and risk of contacting the pleura were more favorable with the greater arm abduction. We conclude that when the arm is abducted to 90 degrees, a 65 degrees -needle angle to the skin appears optimal for contacting the cords, still with a minimal risk of penetrating the pleura. However, this needs to be confirmed by a clinical study. PMID- 11004052 TI - Hyaluronidase as an adjuvant in bupivacaine-lidocaine mixture for retrobulbar/peribulbar block. AB - Hyaluronidase 7.5 IU/mL added to the local anesthetic improves peribulbar block, but smaller concentrations have not been shown to be effective. In this prospective, double-blinded study, 714 consecutive ocular surgery patients were randomized into three groups: no hyaluronidase (n = 241), hyaluronidase 3.75 IU/mL (n = 244), and hyaluronidase 7.5 IU/mL (n = 229). Retrobulbar/peribulbar block was performed with two injections of a 1:1 mixture of bupivacaine 0.75% and lidocaine 2%, 6-8 mL. Patient data were collected on demographics, initial volume of local anesthetic, need for supplementary block, and akinesia of the anesthetized eye. When hyaluronidase was used (3.75 or 7.5 IU/mL), the initial block was sufficient and the anesthetized eye was akinetic significantly more often than in the group without hyaluronidase. The hyaluronidase groups (3.75 and 7.5 IU/mL) did not differ significantly in any respect. We conclude that the addition of hyaluronidase 3.75 or 7.5 IU/mL improved the success of the initial retrobulbar/peribulbar block and akinesia and reduced the need for supplementary block. IMPLICATIONS: We conclude that the addition of hyaluronidase 3.75 or 7.5 IU/mL improved the success of the initial retrobulbar/peribulbar block and akinesia and reduced the need for supplementary block. PMID- 11004053 TI - A comparative study with oral nifedipine, intravenous nimodipine, and magnesium sulfate in postoperative analgesia. AB - We tested the ability of two L-type calcium channel blockers (nifedipine and nimodipine) and the N-methyl D-aspartate natural antagonist magnesium to decrease morphine requirements and pain in the postoperative period in 92 patients undergoing elective colorectal surgery. In a randomized, double-blinded study, patients were assigned to one of four groups. The control group received placebo. The nifedipine group received 60 mg of oral nifedipine. The magnesium group received an initial dose of 30 mg/kg followed by 10 mg x kg(-1) x h(-1) of magnesium sulfate over 20 h. The nimodipine group received 30 microg x kg(-1) x h(-1) of nimodipine over 20 h. Postoperative morphine consumption was assessed for 48 h. Pain at rest and pain on movement were assessed up to the fifth day postsurgery. There were no differences among groups in postoperative morphine consumption at 12 and 24 h. The nifedipine group consumed more morphine than the control and nimodipine groups during 24-48 h. Pain at rest scores were higher at 16 and 24 h in the nifedipine group than in the other three groups. Pain on movement scores were lower at 72 h in the nimodipine group than in the control and nifedipine groups. In conclusion, the perioperative application of oral nifedipine, IV nimodipine, or IV magnesium sulfate failed to decrease postoperative morphine requirements after colorectal surgery. IMPLICATIONS: The increase of intracellular calcium plays a key role in spinal transmission of pain and in the establishment of central sensitization. We examined the effects of nifedipine, nimodipine, and magnesium sulfate in postoperative analgesia after colorectal surgery. We found no differences in morphine consumption with the administration of each drug alone. PMID- 11004054 TI - Lidocaine disrupts axonal membrane of rat sciatic nerve in vitro. AB - Highly concentrated lidocaine has been reported to induce irreversible loss of membrane potential in crayfish nerve, which implies membrane disruption as one of the direct mechanisms of lidocaine-induced neurotoxicity. To confirm lidocaine induced membrane disruption in mammalian nerve, a lactate dehydrogenase (LDH) leakage from rat sciatic nerve was measured in vitro. Before applying lidocaine, the desheathed nerve was incubated for 60 min in Krebs-Ringer solution at 37 degrees C to examine basal LDH activity. It was then incubated in 80 mM lidocaine solution at pH 7.3 for 15, 30, 60, or 120 min. Other nerves were immersed in 800 mM choline solution for 120 min. Total LDH activity per wet weight of nerve tissue was assayed using spectrophotometry. It was also determined using nerves cut into 10 segments and incubated in distilled water for 60 min. The LDH activity in the lidocaine group showed a time-dependent increase. After the 60- and 120-min incubation with lidocaine, the amount of LDH activity was significantly increased compared with the choline group and was similar to that of the group incubated in distilled water. We conclude that 80 mM lidocaine may be sufficient to cause membrane damage and facilitate the leakage of enzymes from cytoplasm. IMPLICATIONS: This study demonstrates that exposing the rat myelinated nerve to lidocaine at a clinically used concentration for more than 30 min causes enough membrane damage to allow enzyme leakage. In clinical practice, the smallest effective dose should be used. PMID- 11004055 TI - Percutaneous neuromodulation therapy: does the location of electrical stimulation effect the acute analgesic response? AB - We studied the effect of the location of electrical stimulation on the acute analgesic response to percutaneous neuromodulation therapy in patients with nonradiating neck pain. Sixty-eight patients received three different nonpharmacologic modalities, namely "needles only" (neck), local (neck) dermatomal stimulation, and remote (lower back) dermatomal stimulation in a random sequence over the course of an 11-wk study period. All treatments were given for 30 min, 3 times per week for 3 wk, with 1 wk "off" between each modality. The assessment tools included the health status survey short form (SF 36) questionnaire, as well as 10-cm visual analog scales for assessing pain, physical activity, and quality of sleep. The pain visual analog scale was repeated 5-10 min after each treatment session. The daily oral nonopioid analgesic requirements were recorded in the patient diary during the entire study period. At the end of each 3-wk treatment block, the SF-36 questionnaire was repeated. Compared with needles only and remote dermatomal stimulation, local dermatomal stimulation produced a significantly greater decrease in pain (38%+/ 17% vs 9%+/- 16% and 13%+/-18%), increase in physical activity (41%+/-21% vs 11% +/-17% and 16%+/-15%), and improvement in the quality of sleep (34% +/-18% vs 7%+/-17% and 10%+/-18%) compared with baseline values (P<0.05). The need for oral analgesic medications was decreased by an average of 6%+/-15%, 37%+/-18%, and 9%+/-13% during the 3-wk treatment period with the needle only, local dermatomal, and remote dermatomal stimulation, respectively. The posttreatment SF-36 test results revealed that all three modalities produced improvements compared with the prestudy scores for both the physical component summary and mental component summary. However, the magnitude of the changes in the physical component summary and mental component summary with local dermatomal stimulation was significantly greater (+7.9 and +3.6, respectively) than needle only (+3.4 and +1.7, respectively) or remote dermatomal stimulation (+3.7 and +1.9, respectively). No side effects were reported at the needle insertion sites. We conclude that electrical stimulation at the specific dermatomal levels corresponding to the local pathology produces greater short-term improvements in pain control, physical activity, and quality of sleep in patients with chronic neck pain. PMID- 11004056 TI - The effects of KRN5500, a spicamycin derivative, on neuropathic and nociceptive pain models in rats. AB - We studied the effects of a spicamycin derivative, KRN5500, on two animal models of neuropathic pain (Chung and Bennett models) and a nociceptive pain model by using Complete Freund's adjuvant. After the establishment of mechanical allodynia by using the previously mentioned models, a single intraperitoneal injection of KRN5500 produced significant attenuation of mechanical allodynia in both neuropathic pain models. However, this effect was not observed in rats that had a nociceptive injury (Complete Freund's adjuvant). Furthermore, this experimental drug did not alter the mechanical pain threshold (by using von Frey filament test) on normal, uninjured rats. We have demonstrated that KRN5500 may have value in the treatment of neuropathic pain. PMID- 11004057 TI - Memantine (a N-methyl-D-aspartate receptor antagonist) in the treatment of neuropathic pain after amputation or surgery: a randomized, double-blinded, cross over study. AB - Evidence has accumulated that the N:-methyl-D-aspartate receptor system plays a role in continuous and particularly, in stimulus-evoked pain after nerve injury. We examined, in a randomized, double-blinded, cross-over fashion, the analgesic effect of memantine (a N:-methyl-D-aspartate receptor antagonist) in a group of patients with chronic pain after surgery. We randomized 19 patients to receive either memantine or placebo in the first 5-wk treatment period. A washout period of 4 wks was followed by another 5-wk treatment period with the opposite drug. The dosage of drug was increased from 5 to 20 mg/d. Pain was recorded daily, with the use of a 0-10 numeric rating scale. Before and at the end of each treatment period, pain and sensitivity were also assessed by using the McGill Pain Questionnaire, allodynia to touch, brush and cold, wind-up-like pain, and thresholds to mechanical stimuli (pressure and von Frey hair). A total of 15 patients (12 amputees and three patients with other nerve injuries) completed the study. There was no difference between memantine and placebo on any of the outcome measures. We conclude that memantine at a dosage of 20 mg/d does not reduce spontaneous or evoked pain in patients with nerve injury pain. IMPLICATIONS: In a randomized, double-blinded and cross-over study, the analgesic effect of memantine (a drug which reduces the excitability of sensitized neurons in the dorsal horn) was examined in 19 patients with chronic pain after nerve injury. PMID- 11004058 TI - The effect of systemic lidocaine on pain and secondary hyperalgesia associated with the heat/capsaicin sensitization model in healthy volunteers. AB - Although effective in neuropathic pain, the efficacy of systemic lidocaine in non neuropathic pain remains uncertain. We investigated the analgesic effect of systemic lidocaine on the heat/capsaicin sensitization model of experimental pain in 24 volunteers. Sensitization was produced by heating the skin to 45 degrees C for 5 min, followed by a 30-min application of 0.075% capsaicin cream, and maintained by periodically reheating the sensitized skin. Subjects received IV lidocaine (bolus 2 mg/kg, then infusion 3 mg. kg. h), or saline for 85 min. Areas of secondary hyperalgesia, heat pain detection thresholds, and painfulness of stimulation with 45 degrees C for 1 min (long thermal stimulation) were quantified. Systemic lidocaine reduced the area of secondary hyperalgesia to brush, but not to von Frey hair stimulation. Lidocaine did not alter heat pain detection thresholds or painfulness of long thermal stimulation in normal skin. We conclude that, at infusion rates in the low- to mid-antiarrhythmic range, lidocaine has no effect on acute nociceptive pain but does have a limited and selective effect on secondary hyperalgesia. IMPLICATIONS: The efficacy of systemic lidocaine in nonneuropathic pain remains uncertain. This study investigates the effect of systemic lidocaine on experimental-induced hyperalgesia in 25 volunteers. Hyperalgesia was induced by using an experimental pain model that uses heat and capsaicin in combination. Systemic lidocaine showed a selective effect on secondary hyperalgesia. PMID- 11004059 TI - Defining segments and phases of a time capnogram. AB - The division of a time capnogram into inspiratory and expiratory segments is arbitrary and results in the inability of a time capnogram to detect rebreathing instantaneously. Demarcation of a time capnogram into inspiratory and expiratory components using gas flow signals will not only facilitate prompt detection of rebreathing, but will also allow application of standardized and physiologically appropriate nomenclature for better understanding and interpretation of time capnograms. A Novametrix((R)) CO(2)-SMO plus respiratory profile monitor (Novametrix Medical Systems, Wallingford, CT) was used to obtain a simultaneous display of CO(2) and respiratory flow waveforms on a computer screen during spontaneous and controlled ventilation using a circle system with the inspiratory valve competent (no rebreathing) and with the valve displaced (rebreathing). Because the response time of the CO(2) analyzer was similar to the response time of the flow sensor, a comparison was made between the two waveforms to determine the inspiratory segment (Phase 0) and the expiratory segment of the time capnogram and its subdivisions (Phases I, II, and III). The end of expiration almost coincides with the downslope of the CO(2) waveform in the capnograms when there is no rebreathing. However, in the presence of rebreathing, the alveolar plateau is prolonged and includes a part of inspiration (Phase 0), in addition to the expiratory alveolar plateau (Phase III). IMPLICATIONS: Presently, the division of a time capnogram into inspiratory and expiratory segments is arbitrary. Demarcation of a time capnogram into various components using the gas flow signals facilitates prompt detection of the cause of abnormal capnograms that can widen the scope of future clinical applications of time capnography. PMID- 11004060 TI - Aggressive warming reduces blood loss during hip arthroplasty. AB - We evaluated the effects of aggressive warming and maintenance of normothermia on surgical blood loss and allogeneic transfusion requirement. We randomly assigned 150 patients undergoing total hip arthroplasty with spinal anesthesia to aggressive warming (to maintain a tympanic membrane temperature of 36.5 degrees C) or conventional warming (36 degrees C). Autologous and allogeneic blood were given to maintain a priori designated hematocrits. Blood loss was determined by a blinded investigator based on sponge weight and scavenged cells; postoperative loss was determined from drain output. Results were analyzed on an intention-to treat basis. Average intraoperative core temperatures were warmer in the patients assigned to aggressive warming (36.5 degrees +/- 0.3 degrees vs 36.1 degrees +/- 0.3 degrees C, P< 0.001). Mean arterial pressure was similar in each group preoperatively, but was greater intraoperatively in the conventionally warmed patients: 86+/-12 vs 80+/-9 mm Hg, P<0.001. Intraoperative blood loss was significantly greater in the conventional warming (618 mL; interquartile range, 480-864 mL) than the aggressive warming group (488 mL; interquartile range, 368 721 mL; P: = 0.002), whereas postoperative blood loss did not differ in the two groups. Total blood loss during surgery and over the first two postoperative days was also significantly greater in the conventional warming group (1678 mL; interquartile range, 1366-1965 mL) than in the aggressively warmed group (1,531 mL; interquartile range, 1055-1746 mL, P = 0.031). A total of 40 conventionally warmed patients required 86 units of allogeneic red blood cells, whereas 29 aggressively warmed patients required 62 units (P = 0.051 and 0.061, respectively). We conclude that aggressive intraoperative warming reduces blood loss during hip arthroplasty. IMPLICATIONS: Aggressive warming better maintained core temperature (36.5 degrees vs 36.1 degrees C) and slightly decreased intraoperative blood pressure. Aggressive warming also decreased blood loss by approximately 200 mL. Aggressive warming may thus, be beneficial in patients undergoing hip arthroplasty. PMID- 11004061 TI - Fractional distillation of acid contaminants from sevoflurane. AB - On two occasions, sevoflurane distributed for clinical practice has been found to be contaminated with compounds thought to include hydrogen fluoride (HF) and silicon tetrafluoride (SiF(4)). Both compounds can produce pulmonary injury. However, injury would require fractional distillation of the compounds during the course of sevoflurane vaporization. We hypothesized that such distillation would occur and that the compounds would vaporize more rapidly than would sevoflurane. Thus, we tested whether fractional distillation occurs during vaporization of sevoflurane containing HF or SiF(4), or from sevoflurane containing HF converted to other compounds by contact with glass. Vaporization of < 10% of the sevoflurane distilled 65%-99% of these compounds, SiF(4) distilling most rapidly, HF (converted to other acidic compounds, including SiF(4)) distilling nearly as rapidly, and HF slowest. Nuclear magnetic resonance studies indicated that HF interaction with glass changed all HF to three other compounds, one being SiF(4) and the others being unknown. HF and SiF4 distill from sevoflurane more rapidly than sevoflurane is vaporized. Measurement of acidity after sevoflurane administration may not reveal a previous presence of such contaminants. PMID- 11004062 TI - The epileptogenic properties of the volatile anesthetics sevoflurane and isoflurane in patients with epilepsy. AB - No study comparing epileptogenicity of sevoflurane to other volatile anesthetics has been performed. We compared the epileptogenic properties of sevoflurane to isoflurane in patients with epilepsy. In 24 mentally and/or physically disabled patients, 12 with epilepsy and 12 without epilepsy, electroencephalograms were recorded under anesthesia with 1.0 minimum alveolar anesthetic concentration (MAC), 1.5 MAC, and then 2.0 MAC sevoflurane or isoflurane under three ventilatory conditions: (A) 100% oxygen, and end-tidal CO(2) partial pressure (ETCO(2)) = 40 mm Hg, (B) 50% oxygen, 50% nitrous oxide, ETCO(2) = 40 mm Hg, and (C) 100% oxygen, ETCO(2) = 20 mm Hg. Spike activity was evaluated as a spike-and wave index (% durations of spike and wave). The spike-and-wave index increased (P<0.05) from 1.99%+/-0.96% during 1.0 MAC sevoflurane to 6.14% +/- 4.45% during 2.0 MAC sevoflurane in (A) in the epilepsy group, while no spike activity was observed in the nonepilepsy group. Only a few spikes were observed under isoflurane anesthesia, 0.04% +/- 0.04% in (A), with no spikes in (B) and (C). Supplementation with 50% nitrous oxide or hyperventilation (P<0.05) suppressed the occurrence of spikes. Sevoflurane has a stronger epileptogenic property than isoflurane, but nitrous oxide or hyperventilation counteracts this specific epileptogenic property. IMPLICATIONS: The stronger epileptogenicity of sevoflurane than isoflurane was confirmed in a controlled study in patients with epilepsy. Hyperventilation and supplementation of nitrous oxide under sevoflurane anesthesia suppressed epileptogenicity. A combination of sevoflurane and nitrous oxide may be a safer method for seizure-prone patients than the use of sevoflurane alone. PMID- 11004063 TI - Interaction between volatile anesthetics and hypoxia in porcine tracheal smooth muscle. AB - We investigated the direct interaction between the volatile anesthetics, isoflurane and sevoflurane, and hypoxia in porcine tracheal smooth muscle in vitro by simultaneously measuring muscle tension and intracellular concentration of free Ca(2+) ([Ca2+]i). Muscle tension was measured by using an isometric transducer, and [Ca2+]i was measured by using fura-2, an indicator of Ca2+. Under the condition of bubbling with 95% O2/5% CO2, [Ca2+]i was increased by 1 microM carbachol with a concomitant contraction. Volatile anesthetics significantly inhibited both carbachol-induced muscle contraction and increase in [Ca2+]i. Hypoxia bubbled with 95% N(2)/5% CO2 inhibited the muscle contraction by 30% with an increase in [Ca2+]i by 20%. Exposure to hypoxia substantially enhanced the inhibitory effects of these anesthetics on carbachol-induced muscle contraction, whereas the decreases in [Ca2+]i were significantly prevented by hypoxia. Under Ca2+-free conditions, hypoxia significantly decreased the muscle contraction by 20%; however, it still increased [Ca2+]i by 15%. Exposure to the anesthetics significantly enhanced the inhibitory effect of hypoxia on the muscle contraction; however, it appeared to have little effect on [Ca2+]i. Hypoxia inhibits airway smooth muscle contraction independently of intracellular Ca2+, and it substantially potentiates the inhibitory effects of volatile anesthetics on airway smooth muscle contraction. IMPLICATIONS: Hypoxia inhibits agonist induced tracheal smooth muscle contraction with an increase in free Ca2+ [Ca2+]i, which comes from intracellular Ca2+ stores. Hypoxia also potentiates the inhibitory effect of volatile anesthetics on airway smooth muscle contraction. Conversely, there is a possibility that the treatment of asthmatic patients with oxygen partially attenuates the inhibitory effect of volatile anesthetics on airway smooth muscle contractility. PMID- 11004064 TI - A pilot study of left tracheal pulse oximetry. AB - We tested the hypothesis that left tracheal pulse oximetry (SpO(2)) is more accurate than finger SpO(2) when compared with oxygen saturation from arterial blood samples (SaO(2)) in anesthetized patients with normal thoracic anatomy. We also tested the hypothesis that tracheal oximetry readings are primarily derived from the tracheal mucosa. We studied 20 hemodynamically stable, well oxygenated, anesthetized patients with normal anatomy (ASA physical status I-III, 18-80 yr old). A single-use pediatric pulse oximeter was attached to the left lateral surface of a tracheal tube cuff. Tracheal and finger SpO(2) (dominant index finger), and SaO(2) (nondominant radial artery) were taken with the intracuff pressure at 0-60 cm H(2)O. Tracheal SpO(2) was the same as SaO(2) at an intracuff pressure of 10-60 cm H(2)O, but was less when the intracuff pressure was zero (P<0.0001). Tracheal SpO(2) was higher than finger SpO(2) at an intracuff pressure of 10-60 cm H(2)O (all: P <0.001), but was lower when the intracuff pressure was zero (P< 0.0001). SaO(2) was always higher than finger SaO(2) (P<0.0001). Tracheal SpO(2) was lower at an intracuff pressure of zero (P< 0.0001), but was otherwise similar over the range of intracuff pressures. SaO(2) and finger SpO(2) did not vary with intracuff pressure. Tracheal SpO(2) agrees more closely with SaO(2) than finger SpO(2) at an intracuff pressure of 10-60 cm H(2)O (mean difference < 0.2%). We conclude that left tracheal SpO(2) is feasible and provides similar readings to arterial blood samples and more accurate readings than finger oximetry in hemodynamically stable, well oxygenated, anesthetized patients with normal thoracic anatomy. Tracheal oximetry readings are not primarily derived from the tracheal mucosa. The technique merits further evaluation. PMID- 11004065 TI - Effects of nonimmobilizers and halothane on Caenorhabditis elegans. AB - We studied the effects of two nonimmobilizers, a transitional compound, and halothane on the nematode, Caenorhabditis elegans, by using reversible immobility as an end point. By themselves, the nonimmobilizers did not immobilize any of the four strains of animals tested. Toluene appears to be a transitional compound for all strains tested. The additive effects of the nonimmobilizers with halothane were also studied. Similar to results seen in studies of mice, the nonimmobilizers were antagonistic to halothane in the wild type nematode. However, the nonimmobilizers did not affect the 50% effective concentrations of halothane for two other mutant strains. For halothane, the slopes of the dose response curves were smaller in more sensitive strains compared with the wild type. As in mammals, nonimmobilizers antagonize the effects of halothane on the nematode, C. elegans. The variation in slopes in the response to halothane in different strains is consistent with multiple sites of action. These results support the use of C. elegans as a model for the study of anesthetics. PMID- 11004066 TI - The nonimmobilizer 1,2-dichlorohexafluorocyclobutane does not affect thermoregulation in the rat. AB - Inhaled and other anesthetics profoundly affect the central nervous system, causing amnesia, immobility in the face of noxious stimulation, and depression of thermoregulation. Nonimmobilizers, inhaled compounds whose lipophilicity suggests that they should be anesthetics, do not produce immobility, but they do cause amnesia. Their effects on thermoregulation were the subject of the present study. We gave eight rats isoflurane on one occasion and the nonimmobilizer 2N (1,2 dichlorolhexafluorocyclobutane) on another. We measured the effect of various concentrations of each compound on thermoregulation provoked by body cooling. The specific outcome was increased metabolism, as reflected in increased output of carbon dioxide. Isoflurane decreased the temperature threshold for such increases and the maximum response intensity, doing so in a concentration-dependent manner, whereas 2N had a minimal or no effect at any concentration up to 0.9 minimum alveolar concentration (estimated from its lipophilicity). Thus, 2N may be a useful tool for studies of the mechanisms mediating the thermoregulatory depression produced by anesthetics: 2N should not affect such a mechanism. PMID- 11004067 TI - Does the ProSeal laryngeal mask airway prevent aspiration of regurgitated fluid? AB - In this randomized, cross-over cadaver study, we determined whether a new airway device, the ProSeal laryngeal mask airway (PLMA; Laryngeal Mask Company, Henley on-Thames, UK), prevents aspiration of regurgitated fluid. We studied five male and five female cadavers (6-24 h postmortem). The infusion set of a pressure controlled, continuous flow pump was inserted into the upper esophagus and ligated into place. Esophageal pressure (EP) was increased in 2-cm H(2)O increments. This was performed without an airway device (control) and over a range of cuff volumes (0-40 mL) for the classic laryngeal mask airway (LMA), the PLMA with the drainage tube clamped (PLMA clamped) and unclamped (PLMA unclamped). The EP at which fluid was first seen with a fiberoptic scope in the hypopharynx (control), above or below the cuff, or in the drainage tube, was noted. Mean EP at which fluid was seen without any airway device was 9 (range 8 10) cm H(2)O. EP at which fluid was seen was always higher for the PLMA clamped and LMA compared with the control (all, P<0.0001). The mean EP at which fluid was seen for the PLMA unclamped was similar to the control at 10 (range 8-13) cm H(2)O. For the PLMA unclamped, fluid appeared from the drainage tube in all cadavers at 10-40 mL cuff volume and in 8 of 10 cadavers at zero cuff volume. Mean EP at which fluid was seen above the cuff was similar for the PLMA clamped and LMA at 0-30 mL cuff volume, but was higher for PLMA clamped at 40-mL cuff volume (81 vs 48 cm H(2)O, P = 0.006). Mean EP at which fluid was seen below the cuff was similar at 0-10 mL cuff volume, but was higher for the PLMA clamped at 20, 30, and 40 mL cuff volume (62, 68, 73 vs. 46, 46, 46 cm H(2)O, respectively, P<0.04). For the PLMA clamped and the LMA, fluid appeared simultaneously above and below the cuff at all cuff volumes. We concluded that in the cadaver model, the correctly placed PLMA allows fluid in the esophagus to bypass the pharynx and mouth when the drainage tube is open. Both the LMA, and PLMA with a closed drainage tube, attenuate liquid flow between the esophagus and pharynx. This may have implications for airway protection in unconscious patients. PMID- 11004068 TI - A modified intubating laryngeal mask for endotracheal tube exchange. AB - IMPLICATIONS: It is often necessary to change a patient's breathing tube (endotracheal tube). This can be a risky procedure. This report describes a technique for changing an endotracheal tube by using a modified "intubating laryngeal mask" (a commonly used airway and breathing device) and a fiberoptic bronchoscope. PMID- 11004069 TI - The efficacy and safety of EMLA cream for awake fiberoptic endotracheal intubation. AB - EMLA Cream (EC; Astra, Westborough, MA) has been widely used as a local anesthetic. Limited safety information is available with respect to the application of EC to the oral mucous membranes. The purpose of this pilot study was to evaluate the efficacy and safety of EC when applied to oral mucosa for fiberoptic intubation. Twenty ASA physical status I-IV patients (11 women and 9 men), 28-57 yr old, who were scheduled for awake, fiberoptic, intubation participated in this open-label study. A total of 4 g of EC was used for 5 min until the patient showed no evidence of a gag reflex (this was evaluated clinically by the patient's acceptance of the William's airway and considered the endpoint for assessing adequate topicalization of the oropharynx). The measured peak plasma concentration of lidocaine or prilocaine did not reach toxic levels in any patient. Methemoglobin levels did not exceed normal values (1.5%) in any patient, and there was no relationship between methemoglobin levels and patient weight, amount of EC used, measured peak plasma concentration, or times to measured peak concentrations of prilocaine or lidocaine. We conclude that EC provided satisfactory topical anesthesia allowing for successful oral fiberoptic intubation in all patients and should be considered a safe alternative for anesthetizing the airway of patients requiring awake oral fiberoptic intubation. PMID- 11004070 TI - Bacitracin irrigation: a cause of anaphylaxis in the operating room. AB - IMPLICATIONS: We report a unique case of acute anaphylaxis after mediastinal irrigation with a dilute bacitracin solution. PMID- 11004071 TI - The anesthetic management of a patient with paroxysmal nocturnal hemoglobinuria. AB - IMPLICATIONS: This case report describes the anesthetic considerations for a patient with paroxysmal nocturnal hemoglobinuria. Specific strategies to be applied in the perioperative period to prevent hemolytic episodes and venous thrombosis are also discussed. PMID- 11004072 TI - An unusual case of malignant hyperthermia during desflurane anesthesia in an African-American patient. AB - IMPLICATIONS: Malignant hyperthermia is an uncommon, heritable condition triggered by anesthesia and is followed by an increase in temperature that may be fatal without prompt treatment. It is rare with desflurane and in black individuals of African descent. We present a case of malignant hyperthermia in an African-American patient during desflurane anesthesia. PMID- 11004073 TI - The effect of sedative drugs on diaphragmatic contractility in dogs: propofol versus midazolam. AB - IMPLICATIONS: a sedative dose (0.1 mg x kg(-1) x h(-1)) of midazolam, compared with a subhypnotic dose (1.5 mg x kg(-1) x h(-1)) of propofol, decreases the contractility of the diaphragm in dogs. PMID- 11004074 TI - Beware of air in the blood pump. PMID- 11004075 TI - Muscle relaxants and 5-HT(3) receptors. PMID- 11004076 TI - Inhibition of epidural morphine-induced pruritus by epidural droperidol. PMID- 11004077 TI - No need for claims: facts rule performance of jet ventilation. PMID- 11004078 TI - Sticker sedation. PMID- 11004079 TI - Nitric oxide delivery system. PMID- 11004080 TI - New cuff design prevents N2O diffusion. PMID- 11004081 TI - NMDA receptor blockade: from the laboratory to clinical application. PMID- 11004082 TI - Intravenous lidocaine and intrathecal opioids. PMID- 11004083 TI - More about the Hawthorne effect. PMID- 11004084 TI - Presence of placental tissue is necessary for TNG to provide uterine relaxation. PMID- 11004085 TI - An unusual cause of failed sedation. PMID- 11004086 TI - Teaching fiberoptic intubation in the pediatric patient. PMID- 11004087 TI - Hypercapnea is an important adjuvant factor of oculocardiac reflex during strabismus surgery. PMID- 11004088 TI - Matrix metalloproteinases and their natural inhibitors. PMID- 11004089 TI - The conundrum of sweet hyperopia. PMID- 11004090 TI - Matrix metalloproteinases and their natural inhibitors in fibrovascular membranes of proliferative diabetic retinopathy. AB - AIM: To examine epiretinal membranes of proliferative diabetic retinopathy (PDR) for the presence of selective matrix metalloproteinases (MMPs) and their natural inhibitors (TIMPs), in order to determine whether neovascularisation and fibrosis, characteristic of this complication of diabetes mellitus, are associated with specific anomalies of MMP or TIMP expression. METHODS: The presence of selected MMPs and TIMPs was investigated in 24 fibrovascular epiretinal membranes of PDR, and the findings compared with that observed in 21 avascular epiretinal membranes of proliferative vitreoretinopathy (PVR) and five normal retinas. Specimens were examined for deposition of interstitial collagenase (MMP-1), stromelysin-1 (MMP-3), gelatinase A (MMP-2), gelatinase B (MMP-9), and three tissue inhibitors of metalloproteinases (TIMP-1, TIMP-2, and TIMP-3). RESULTS: The results showed that unlike normal retina, which constitutively expresses MMP-1 and TIMP-2, a large proportion of PDR membranes (> 62%) stained for MMP-1, MMP-2, MMP-3, MMP-9, TIMP-1, TIMP-2, and TIMP-3. There were no differences in the expression of these molecules when compared with PVR membranes. A characteristic staining for MMP-9 was observed within the perivascular matrix of PDR membranes, and there was a significant increase in TIMP-2 expression by PDR membranes (p= 0.036) when compared with PVR membranes. CONCLUSIONS: The findings that MMPs involved in degradation of fibrovascular tissue matrix, as well as TIMP-1 and TIMP-2, are found in a large proportion of PDR membranes, and that their expression does not differ from that of PVR membranes, suggest the existence of common pathways of extracellular matrix degradation in pathological processes leading to retinal neovascularisation and fibrosis. PMID- 11004091 TI - Refractive changes in diabetic patients during intensive glycaemic control. AB - AIMS: To evaluate the clinical course and the characteristics of transient refractive error occurring during intensive glycaemic control of severe hyperglycaemia. METHODS: 28 eyes of patients with persistent diabetes were included in this prospective study. During the observation period, patients underwent general ophthalmological examination and A-mode scan ultrasonography was performed at each examination-at days 1, 3, and 7, and then once every week or every other week until recovery of hyperopia. RESULTS: A transient hyperopic change occurred in all patients receiving improved control after hyperglycaemia. Hyperopic change developed a mean of 3.4 (SD 2. 0) days after the onset of treatment, and reached a peak at 10.3 (6. 1) days, where the maximum hyperopic change in an eye was 1.47 (0. 87) D (range 0.50-3.75 D). Recovery of the previous refraction occurred between 14 and 84 days after the initial assessment. There was a positive correlation between the magnitude of the maximum hyperopic change and (1) the plasma glucose concentration on admission (p<0.01), (2) the HbA(1c) level on admission (p<0.005), (3) the daily rate of plasma glucose reduction over the first 7 days of treatment (p<0.001), (4) the number of days required for hyperopia to reach its peak (p<0.001), and (5) the number of days required for the development and resolution of hyperopic changes (p<0.0001). There was a negative correlation between the maximum hyperopic change of an eye and baseline value of refraction (p<0.01). During transient hyperopia, no significant changes were observed in the radius of the anterior corneal curvature, axial length, lens thickness, or depth of anterior chamber. CONCLUSIONS: The degree of transient hyperopia associated with rapid correction of hyperglycaemia is highly dependent on the rate of reduction of the plasma glucose level. A reduction of refractive index in intraocular tissues, especially in lens, appears to be responsible for this hyperopic change. PMID- 11004092 TI - Patient characteristics, diagnosis, and treatment of non-contact lens related Acanthamoeba keratitis. AB - AIM: To review the clinical characteristics, diagnosis, and visual outcome in patients with non-contact lens related Acanthamoeba keratitis and compare the findings with reported series of contact lens associated Acanthamoeba keratitis. METHODS: Medical and microbiology records of 39 consecutive patients with a diagnosis of Acanthamoeba keratitis, at a tertiary eyecare centre in India between January 1996 and June 1998, were analysed retrospectively. RESULTS: A majority of the patients presented with poor visual acuity and large corneal stromal infiltrates (mean size 38.20 (SD 26. 18) mm). A predisposing factor was elicited in 19/39 (48.7%) patients (trauma 15, dirty water splash three, leaf juice one). None of the patients had worn contact lenses. Most patients (26/39 (66. 6%)) came from a low socioeconomic background. Complaint of severe pain was not a significant feature and radial keratoneuritis was seen in 1/39 (2.5%) patients. A ring infiltrate was present in 41.1% of cases. A clinical diagnosis of fungal keratitis was made in 45% of the patients before they were seen by us. However, all patients were diagnosed microbiologically at our institute based on demonstration of Acanthamoeba cysts in corneal scrapings (34/39) and/or culture of Acanthamoeba (34/39). Treatment with biguanides (PHMB, 15/38 (39.4%), PHMB with CHx, 23/38 (60.5%), one patient did not return for treatment) resulted in healing with scar formation in 27 out of 31(87.0%) followed up patients (mean time to healing 106.9 days). Overall visual outcome was poor with no statistical difference between cases diagnosed within 30 days (early) or 30 days after (late) start of symptoms. The visual outcome in cases requiring tissue adhesive (five) and keratoplasty (three) was also poor. CONCLUSIONS: This is thought to be the largest series of cases of Acanthamoeba keratitis in non-contact lens wearers. In such cases, the disease is advanced at presentation in most patients, pathognomonic clinical features are often not seen, disease progression is rapid, and visual outcome is usually poor. Possible existence of Acanthamoeba pathotypes specifically associated with non-contact lens keratitis and unique to certain geographical areas is suggested. PMID- 11004093 TI - Contrast sensitivity and glare disability by halogen light after monofocal and multifocal lens implantation. AB - BACKGROUND: Standard examination of contrast sensitivity under conditions of glare disability is performed with incandescent light. A new halogen glare test that simulates glare as seen with oncoming vehicle headlights was used to measure glare disability in patients implanted with multifocal and monofocal intraocular lenses (IOLs). METHODS: 28 patients with an average age of 69 years (SD 12 years) were implanted with a monofocal IOL (SI-40NB, Allergan) and 28 patients with an average of 66 years (12 years) were implanted with a refractive multifocal IOL (Array-SA-40N, Allergan). All patients were followed for 5 months postoperatively. Contrast sensitivity at four spatial frequencies (3, 6, 12, and 18 cycles per degree, cpd) with and without a glare source were measured using the halogen glare test (CSV-1000 HGT). Statistical analysis was performed using the two sample Wilcoxon test. The local significance level was set at 0.05. RESULTS: When tested at the lowest spatial frequency (3 cpd) without halogen glare, contrast sensitivity was lower in the multifocal group than in the monofocal group (p=0.0292). With additional glare, there was no difference between both groups. At all other spatial frequencies (6, 12, and 18 cpd), when tested without halogen glare (6 cpd, p=0.5250; 12 cpd, p=0.8483; 18 cpd, p=0.9496) and with moderate (3 cpd, p=0.7993; 6 cpd, p=0.4639; 12 cpd, p=0.7456; 18 cpd, p=1.0) and high halogen glare (3 cpd, p=0. 1513; 6 cpd, p=0.2016; 12 cpd, p=0.3069; 18 cpd, p=0.9933), there was no statistically significant difference between groups. Patients in both groups of age 70 or older had reduced contrast sensitivity without halogen glare and with moderate and strong glare. When monofocal and multifocal patients older than 70 years of age were analysed separately, there was no statistically significant difference in contrast sensitivity with and without glare. Astigmatism >1 dioptre had no significant influence on contrast sensitivity and glare disability when monofocal and multifocal eyes were compared. CONCLUSION: Reduced contrast sensitivity was found in the multifocal group only at the lowest spatial frequency without halogen glare. The monofocal and multifocal groups had no statistically significant differences in contrast sensitivity with moderate and strong glare. These results suggest no difference in glare disability induced by halogen light similar to oncoming vehicle headlights for patients implanted with monofocal and multifocal IOLs. PMID- 11004094 TI - Anterior chamber depth measured by two methods in myopic and hyperopic phakic IOL implant. AB - AIM: To evaluate the accuracy and reliability of the optical versus ultrasonic measurement of anterior chamber depth (ACD) in a group of patients affected by high myopia or hyperopia, before phakic IOL implant. METHODS: 18 consecutive patients (34 eyes) were enrolled in this study, and asked to undergo phakic IOL implant to correct high myopia or hyperopia. The mean age was 29.5 (SD 3.4) years, the male/female ratio was 10/8. 13 patients (24 eyes) were myopic (mean myopia -16.17 (4.39) D, range -9 to -24 D), whereas five patients (10 eyes) were hyperopic (mean hyperopia 7.4 (2.01) D, range 5.5-11 D). For each patient, ACD was evaluated comparing an optical system (Orbscan topograph system) with a standard A-scan ultrasound system. To evaluate the reliability of the two methods, the average of three optical and 10 ultrasonic consecutive measurements were considered. Statistical analysis was performed by means linear regression. RESULTS: The average difference between optical and ultrasound values was 0.17 (0.1) mm (4.68% (2.52%)). There was a constant underestimation of the ACD with the optical system compared with the ultrasound measurement, more evident in the hyperopic eyes (5.20% (1. 95%)) than in myopic ones (4.46% (2.72%)). The reliability of measurements, as showed by the standard deviation values, was higher in the optical system (0.03 (0.02)) than in the ultrasound (0.1 (0. 05)), with a statistical difference (p<0.001). The linear regression analysis between optical and ultrasound measurements was 0.8992 (p<0. 0001). CONCLUSIONS: The optical measurement of ACD is an accurate and reliable technique in high myopic and hyperopic eyes. It gives a small underestimation of the ACD values, but it could be preferable to the ultrasound technique, because it demonstrates more repeatability and has the advantages of a non-contact technique. PMID- 11004095 TI - Quantitative measurement of the PCCC area in the postoperative period. AB - BACKGROUND/AIMS: The major complication of extracapsular cataract extraction (ECCE) is posterior capsule opacification (PCO). Posterior continuous circular capsulorhexis (PCCC) seems to be very promising in preventing PCO. This study was aimed at determining if the PCCC area changes as a function of time and if pearl formation could influence it. METHODS: 24 eyes of 23 patients underwent ECCE with PCCC. Retroillumination photographs were taken at 6 months and then yearly. To measure the PCCC area, the computerised program EPCO (evaluation of posterior capsule opacification) was used. The ratio of the PCCC area in relation to the IOL surface was calculated for the different time stages and the presence of pearl formation was noted. Firstly, proportional changes in diameter were compared in PCCC areas measured after 6 months and after 1 year (group I, n=13) and after 1 year and 2 years (group II, n=14). Secondly, PCCC areas were compared between two time stages in patients with (group III, n=19) and without pearl formation (group IV, n=8). RESULTS: No statistically significant difference was found in diameter change in either group. The PCCC area remains stabile between 6 months to 1 year and 1 year to 2 years. No differences are found between eyes with or without pearl formation. CONCLUSIONS: The PCCC area remains stable as a function of time and is not influenced by pearl formation. PMID- 11004096 TI - Anterior uveitis and its relation to stress. AB - BACKGROUND/AIMS: Idiopathic recurrent acute anterior uveitis (RAAU) is a common reason for attendance at ophthalmic casualty departments. Patients who suffer with this condition may have multiple recurrent episodes which are often debilitating and necessitate time off work. If recurrences are identified early and treatment initiated rapidly, the inflammation can be minimised and settles quickly on topical treatment with a speedy functional recovery by the patient. It has been our clinical impression that some patients with RAAU relate recurrences of inflammation to psychological factors. Therefore stress could be a trigger factor for recurrence, and at such times, patients should be alert to the early signs of uveitis and seek prompt examination by an ophthalmologist. This study tests the hypothesis that stressful life events are associated with the onset of RAAU. METHODS: The number of stressful life events in patients presenting to the casualty department with RAAU in the month immediately preceding symptom onset was compared with a control group of patients with other ocular problems taken from the casualty department using the Holmes and Rahe scale and the Spielberger state-trait anxiety inventory. RESULTS: 35 patients with RAAU did not report a higher number of stressful life events or higher levels of anxiety compared with 29 controls in this study on both Holmes and Rahe scale and Spielberger state trait anxiety inventory. Scores for both groups were comparable with those published for normal adults of the same age and sex. CONCLUSION: Stress was not shown in this study to be involved in triggering the recurrence of idiopathic acute anterior uveitis. PMID- 11004097 TI - Bartonella henselae associated uveitis and HLA-B27. AB - AIM: To investigate the frequency of HLA-B27 in patients with presumed Bartonella henselae associated uveitis and to describe the clinical characteristics of HLA B27 positive patients with uveitis and presumed ocular bartonellosis (POB). METHODS: The diagnosis of POB was considered in 19 patients with unexplained uveitis (except for the HLA-B27 association) and high positive IgG (titre >/=1:900) and/or IgM (titre >/=1:250) antibodies against B henselae. In addition to B henselae serology and HLA-B27 typing, all patients underwent an extensive standard diagnostic screening procedure for uveitis and in all cases the results were within the normal limits. The control group consisted of 25 consecutive patients with panuveitis and negative B henselae serology. RESULTS: HLA-B27 was positive in six of the 19 patients (32%) with POB in contrast to the 4% frequency of HLA-B27 in the control group (p=0.03) and 8% prevalence of HLA-B27 in the Dutch population (p=0.003). At the time of positive Bartonella serological testing five of six HLA-B27 positive patients with POB had severe posterior segment involvement with papillitis, macular oedema, and vitreitis. The duration of intraocular inflammatory activity was more than 6 months in five HLA-B27 positive patients. Although four of the six HLA-B27 positive patients had previous recurrent attacks of acute anterior uveitis, the clinical presentation at the time of positive Bartonella serology differed, as illustrated by the involvement of the posterior segment and chronic course of the ocular disease. CONCLUSIONS: The frequency of HLA-B27 in patients with uveitis and serological characteristics of acute infection with B henselae is higher than in the general Dutch population. The findings of this study also suggest a relation between infection with Bartonella species and HLA-B27. PMID- 11004098 TI - Soluble Fas ligand and soluble Fas in ocular fluid of patients with uveitis. AB - AIMS: To investigate the presence of soluble Fas ligand (sFasL) and soluble Fas (sFas) in ocular fluid of patients with uveitis. METHODS: Samples of aqueous humour (AH, n=17), vitreous fluid (n=9), and serum (n=60) were collected from patients with uveitis which included Behcet's disease, Vogt-Koyanagi-Harada disease, sarcoidosis, human T lymphotropic virus type 1 (HTLV-I) uveitis, sympathetic ophthalmia, HLA-B27 associated acute anterior uveitis, and ocular toxoplasmosis. The AH of patients with age related cataract without uveitis obtained during cataract surgery was used as controls (n=20). The amounts of sFasL and sFas were measured by enzyme linked immunosorbent assay. RESULTS: Significant amounts of sFasL were detected in AH of patients with age related cataract (non-uveitis group). sFasL was also detected in AH of patients with uveitis, though the amounts were slightly lower than those in the non-uveitis group. On the other hand, the levels of sFas in AH of patients with uveitis were significantly higher than those in controls. As for the disease activity, the levels of sFasL and sFas in the vitreous fluid of patients with active uveitis were significantly higher than those in inactive uveitis. sFasL in the serum of healthy donors and patients with uveitis was below detectable levels, except for patients with HTLV-I uveitis who had significant amounts of sFasL in the serum. The levels of sFas in the serum of patients with Behcet's disease, sarcoidosis, and HTLV-I uveitis were significantly higher than those of healthy donors. CONCLUSIONS: sFasL is present in the AH of non-uveitic eyes with age related cataract. Intraocular levels of sFasL and sFas are significantly increased in uveitis, particularly in active uveitis. These data suggest that intraocular sFasL and sFas may have a regulatory role in uveitis. PMID- 11004099 TI - Spontaneous cataract absorption in patients with leptospiral uveitis. AB - AIMS: To describe the occurrence of spontaneous cataract absorption in patients with leptospiral uveitis. METHODS: The records of patients with seropositive leptospiral uveitis seen in the uveitis clinic at Aravind Eye Hospital between January 1994 and December 1997 were reviewed retrospectively. RESULTS: During the 4 years of the study, 394 eyes of 276 patients with seropositive leptospiral uveitis were identified. Of these, 54 eyes (13.7%) of 41 patients (14.9%) had a final visual acuity of 20/40 or worse attributable to cataract formation. Of these 54 eyes, 41 eyes (75.9%) had visually significant cataract on their first visit to the uveitis clinic, and 13 eyes (24.1%) were noted to have cataract 1-6 months after presentation. Spontaneous absorption was observed in 10 eyes (18.5%) of eight patients (19.5%), and occurred from 6 weeks to 18 months, with a median of 5 months, after the onset of cataract. Of 12 035 consecutive, non-leptospiral, non-traumatic, uveitic, control patients seen during the same 4 years of the study, none showed spontaneous cataract absorption. CONCLUSION: Spontaneous cataract absorption occurs in a significant number of patients with leptospiral uveitis, and appears to be unique to this form of non-traumatic uveitis. PMID- 11004100 TI - Ultrasound biomicroscopic analysis of pseudophakic pupillary block glaucoma induced by Soemmering's ring. AB - AIM: To perform ultrasound biomicroscopic analysis of pseudophakic pupillary block glaucoma induced by lens capsule and a Soemmering's ring and its resolution, and to elucidate the pathophysiology of this glaucoma. METHODS: A woman with pseudophakic pupillary block glaucoma underwent successful neodymium:YAG (Nd:YAG) laser photodisruption of the lens capsule through a laser iridotomised coloboma with resolution of the pupillary block. The Humphrey ultrasonic biomicroscope model 840 was employed to observe the anterior segment before and after laser photodisruption. RESULTS: Ultrasound biomicroscopic examination revealed the intraocular lens (IOL) was displaced forward, shallowing the central anterior chamber. The anterior hyaloid face was relatively posterior to the posterior capsule. The iris was in apposition to the anterior capsule, which was in touch with the IOL optics. A massive Soemmering's ring, which extended from the IOL optics to the ciliary processes, was displaced anteriorly. The Soemmering's ring consisted of several tightly packed layers. The ciliary processes rotated anteriorly. After Nd:YAG laser photodisruption of the capsule, ultrasound biomicroscopic images showed the resolution of the pupillary block and the anterior rotation of the ciliary process. The Soemmering's ring moved posteriorly, and the layers in the ring became loose. CONCLUSIONS: A massive Soemmering's ring may one of potential causes of pupillary block after cataract surgery. Ultrasound biomicroscopy is potentially useful as a non-invasive diagnostic technique for clinical diagnosis and differentiation between Soemmering's ring induced pupillary block glaucoma and other forms of pupillary block. PMID- 11004101 TI - Pilot study of the multifocal electroretinogram in ocular hypertension. AB - AIMS: To investigate the variation of retinal function in ocular hypertension (OHT) using the multifocal electroretinogram (MERG), that can objectively assess regional retinal responses using kernel analysis. METHODS: Patients with OHT were recruited for 61 flash MERG recording. The first and second order kernel responses were analysed and compared with responses from a normal group. All the patients with OHT had full eye examinations and visual field analysis to ensure they had no ocular pathology, apart from high IOP (>/= 22 mm Hg). RESULTS: In OHT, both the first and the second order kernel responses showed a reduction in magnitude compared with control values. The second order kernel responses showed larger relative reduction of amplitude than the first order responses. In addition, the macula showed a greater reduction in response than the periphery. CONCLUSIONS: First order and second order kernel analyses are useful for detecting changes of retinal response in OHT. The second order kernel analysis is important in measuring the inner retinal activity and is an important factor in detecting the early glaucoma case. Diminished macular response may be a useful sign in early glaucomatous changes. PMID- 11004102 TI - Comparison of visual field progression in patients with normal pressure glaucoma between eyes with and without visual field loss that threatens fixation. AB - AIM: To compare the frequency and site of visual field progression and changes in visual acuity in patients with normal pressure glaucoma (NPG) with and without pre-existing visual field loss. METHOD: Patients with normal tension glaucoma were selected who had at least 10 visual fields over 5 or more years of follow up and no other condition that might influence the visual field or visual acuity. Alternate left and right eyes were selected from patients in random order. These eyes were then subdivided according to visual field defect threatening fixation, visual field defect not threatening fixation, and no visual field defect (fellow eyes). Eyes were defined as showing a threat to fixation according to the presence of a visual field defect involving one of more of four paracentral visual field locations. Pointwise linear regression analysis was applied to each visual field series using PROGRESSOR software. Progression of visual field loss was defined as the appearance of a regression slope 1 dB per year or more with a significance of p<0.01, which remained consistent with the addition of two of three successive visual fields to the series. The number of patients showing progression and the number where progression occurred in one of the four paracentral visual field locations was noted. The number of eyes losing two or more lines of Snellen visual acuity over the follow up period was also noted. RESULTS: 174 eyes of 174 patients were selected. 106 eyes had visual field loss threatening fixation, 46 eyes had visual field loss that did not threaten fixation, and 22 were fellow eyes with normal visual fields. The median follow up was 7.2 years. Eight eyes (36.4%) in the "normal visual fields" group, 31 eyes (67.4%) in the "visual field loss away from fixation" group, and 87 eyes (82.1%) in the "threat to fixation" group showed progression in any part of the visual field. Two eyes (9.1%) in the "normal visual fields" group, nine eyes (19.6%) in the "visual field loss away from fixation" group, and 45 eyes (42.5%) in the "threat to fixation" group showed progression at "threat to fixation". The Cox proportional hazards regression model showed an increased risk of progression at any part of the visual field for female sex and a decreased risk for eyes with normal visual fields. For progression at threat to fixation this model showed an increased risk with pre-existing threat to fixation. Eyes from older patients and those that went on to have progressive visual field loss at fixation were more likely to lose two lines of Snellen visual acuity over the follow up period. CONCLUSION: Since 20-30% of previously field damaged eyes and over 60% without prior field loss fail to demonstrate progressive visual field damage over a long follow up it is recommended that normal pressure glaucoma patients be monitored for progression and that potentially harmful therapy be withheld until progression is demonstrated. Although the presence of visual field loss that threatens fixation does not constitute an increased risk of visual field progression it does indicate an increased risk of further loss of visual field close to fixation which is in turn associated with loss of central acuity. In the light of this finding, patients with visual field loss that threatens fixation should be managed more aggressively. PMID- 11004103 TI - Utility of predictive equations for estimating the prevalence and incidence of primary open angle glaucoma in the UK. AB - AIMS: To assess the Tuck-Crick and the Quigley-Vitale predictive regression equations against fresh independent real world data for estimating the prevalence of primary open angle glaucoma (POAG) in the UK. To apply the equations to the elderly population of England and Wales, for which there is sample survey data on glaucoma, and demographic data. METHODS: Directly measured actual prevalence of POAG in a population sample of 1530 people was compared with the predicted prevalence derived by applying the Tuck-Crick and the Quigley-Vitale equations to the sample data. The two equations were applied to the demographic data of the population to project POAG prevalence and to derive 5 year cumulative incidence for the elderly population. These were compared with estimates derived from the local survey data. RESULTS: The actual directly measured prevalence of POAG in the local sample was 3.01%. The Tuck-Crick estimate was 2. 74% (difference 0.27%), and the Quigley-Vitale was 3.50% (difference -0.49%). The three methods projection from local survey data, Tuck-Crick, and Quigley-Vitale-gave point estimates of 2.85% (228 526 cases), 2.71% (217 375 cases), and 3.50% (280 364 cases) respectively for the prevalence of POAG in the elderly population of England and Wales (8 008 705 people aged 65 or older). Calculation of incidence from age specific prevalence gave the following results: the numbers of new cases of POAG expected (5 year cumulative incidence) in the elderly population were 71 146 and 94 485 for methods 2 and 3 respectively. CONCLUSIONS: The Tuck-Crick predictive equation performed well when applied to fresh (independent) actual data from a local sample survey using a particular definition for POAG, and its usefulness for estimation of prevalence of POAG in England and Wales has been demonstrated. The work on development of predictive equations has been very promising and further refinements could be made when more fresh survey data become available. PMID- 11004105 TI - At what age could screening for familial retinoblastoma be stopped? A register based study 1945-98. AB - AIM: To evaluate until what age children in families with retinoblastoma should be screened. METHODS: A register based cohort (n= 685) study of Dutch retinoblastoma patients (1945-1998). The records of all familial hereditary retinoblastoma patients from 1945 were reviewed and the age at diagnosis and either they were screened from birth determined. RESULTS: 75 patients had the familial hereditary form of retinoblastoma. The mean age at diagnosis in patients with fundus screening (n=50) from birth on was 4.9 months (median 1.9 months; range 1 day to 48 months). Thus, 4 years was the latest onset of familial retinoblastoma properly evaluated from birth. This mean age was significantly different (p<0.0001) from the mean age at diagnosis in patients without fundus screening (n=25) from birth (mean 17.2 months; median 10.0 months; range 1.5-63.0 months). CONCLUSIONS: Ophthalmological screening of children and sibs at risk for familial hereditary retinoblastoma is recommended until the age of 4 years in order to detect retinoblastoma as early as possible. PMID- 11004104 TI - Vasoproliferative tumours of the retina. AB - BACKGROUND: Vasoproliferative tumours of the retina (VPTR) are benign tumours of unknown origin, occurring mostly in otherwise healthy patients. VPTR may be associated with other chorioretinal diseases, such as uveitis. The tumours, which histologically represent reactive gliovascular proliferations, are characterised by a pink to yellow appearance on funduscopy and are accompanied by exudative and haemorrhagic changes of the retina. METHODS: 22 cases of VPTR in 21 patients were examined with a follow up period between 1 month and 6 years. Ophthalmological changes associated with VPTR were intraretinal and subretinal exudations (n=18), exudative detachments of the surrounding sensory retina (n=13), intraretinal and subretinal haemorrhages (n=10), exudative changes within the macula (n=10), hyperpigmentation of the retinal pigment epithelium at the border of the exudative retinal changes (n=9), and vitreous haemorrhages (n=4). Tumour biopsy was performed in two cases. Treatment consisted of plaque radiotherapy (n=14), plaque radiotherapy and cryotherapy (two), cryotherapy only (two), observation (three), and enucleation in one case of a blind and painful eye. RESULTS: Regression of the tumour and the associated exudative changes could be observed in all treated cases. Visual acuity at last follow up improved two lines or more in two cases, remained within two lines of the initial visual acuity in 15 cases, and worsened in the remaining five. Histopathological examination of the biopsy specimens and the tumour of the enucleated eye showed massive capillary proliferation with perivascular spindle-shaped glial cells of retinal origin. CONCLUSION: The correct diagnosis of VPTR is of importance as these lesions may lead to visual loss. Further, VPTR must be differentiated from angiomas associated with von Hippel-Lindau disease as well as from ocular and systemic malignancies. Regression of tumour thickness and associated retinal changes can be achieved with brachytherapy or cryotherapy. PMID- 11004106 TI - Factors affecting pupil size after dilatation: the Twin Eye Study. AB - BACKGROUND/AIMS: Well dilated pupils make eye surgery easier. A classic twin study was established to examine the relative importance of genes and environment in the variance of pupil size after mydriasis, and to examine the effects of other factors such as age, iris colour, and refractive error. METHODS: 506 twin pairs, 226 monozygotic (MZ) and 280 dizygotic (DZ), aged 49-79 (mean age 62.2 years, SD 5.7) were examined. Dilated pupil size was measured using a standardised grid superimposed over digital retroillumination images taken 50-70 minutes after mydriasis using tropicamide 1% and phenylephrine 10%. Univariate maximum likelihood model fitting was used to estimate genetic and environmental variance components. RESULTS: Dilated pupil size was more highly correlated in MZ compared with DZ twins (intraclass correlation coefficients 0.82 and 0.39 respectively). A model specifying additive genetic and unique environmental factors showed the best fit to the data, yielding a heritability of 78-80%. Individual environmental factors explained 18-19% of the variance in this population. Age only accounted for 2-3% of the variance and refractive error and iris colour did not significantly contribute to the variance. CONCLUSIONS: Pupil size after mydriasis is largely genetically determined, with a heritability of up to 80%. PMID- 11004107 TI - Ocular features in Rubinstein-Taybi syndrome: investigation of 24 patients and review of the literature. AB - AIMS: To delineate the nature and frequency of ocular pathology in Rubinstein Taybi syndrome (RTs). METHODS: Literature was searched for reports describing ocular symptoms in patients with RTs. 24 RTs patients (out of a total of 73 Dutch known RTs individuals) were selected for ophthalmological and electrophysiological examination, selection being based only on the distance between a patient's residence and the place of investigation. RESULTS: Most frequently reported eye anomalies in the literature were lacrimal duct obstruction, corneal abnormalities, congenital glaucoma, congenital cataract, and colobomata. Abnormalities of almost any eye segment have been published in case reports. Ophthalmological examination of 24 Dutch RTs patients showed a visual acuity 70 years, diabetes, smoking, unstable angina, diffuse coronaropathy, and peripheral vasculopathy were all predictors of UAA. Patients were treated with hypothermic ventricular fibrillation (no-touch technique n=129) or beating heart revascularization (no-pump technique n=82) depending on the possibility of founding an arterial cannulation site. The overall in-hospital mortality rate was 2.8% (6 of 211) with no differences between the 2 surgical strategies. The no-touch technique was associated with a greater incidence of neurological complications (stroke and transient ischemic attack), renal insufficiency, and stay in the intensive care unit and hospital. However, at midterm follow-up, more patients of the no-pump group had ischemia recurrence. CONCLUSIONS: A preoperative diagnosis of UAA is achievable only in a minority of patients, which highlights the necessity revising the current diagnostic protocols. The use of the no-touch technique is associated with an high perioperative risk but a superior possibility of complete revascularization, whereas adoption of the no-pump strategy ensures a smoother postoperative course at the expense of an higher incidence of ischemia recurrence. PMID- 11004140 TI - Effect of amlodipine on the progression of atherosclerosis and the occurrence of clinical events. PREVENT Investigators. AB - BACKGROUND: The results of angiographic studies have suggested that calcium channel-blocking agents may prevent new coronary lesion formation, the progression of minimal lesions, or both. METHODS AND RESULTS: The Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial (PREVENT) was a multicenter, randomized, placebo-controlled, double-masked clinical trial designed to test whether amlodipine would slow the progression of early coronary atherosclerosis in 825 patients with angiographically documented coronary artery disease. The primary outcome was the average 36-month angiographic change in mean minimal diameters of segments with a baseline diameter stenosis of 30%. A secondary hypothesis was whether amlodipine would reduce the rate of atherosclerosis in the carotid arteries as assessed with B-mode ultrasonography, which measured intimal-medial thicknesses (IMT). The rates of clinical events were also monitored. The placebo and amlodipine groups had nearly identical average 36-month reductions in the minimal diameter: 0.084 versus 0.095 mm, respectively (P:=0.38). In contrast, amlodipine had a significant effect in slowing the 36-month progression of carotid artery atherosclerosis: the placebo group experienced a 0.033-mm increase in IMT, whereas there was a 0. 0126-mm decrease in the amlodipine group (P:=0.007). There was no treatment difference in the rates of all-cause mortality or major cardiovascular events, although amlodipine use was associated with fewer cases of unstable angina and coronary revascularization. CONCLUSIONS: Amlodipine has no demonstrable effect on angiographic progression of coronary atherosclerosis or the risk of major cardiovascular events but is associated with fewer hospitalizations for unstable angina and revascularization. PMID- 11004141 TI - Explanation for the decline in coronary heart disease mortality rates in Auckland, New Zealand, between 1982 and 1993. AB - BACKGROUND: We sought to determine how much of the recent, substantial fall in coronary heart disease (CHD) mortality rates in New Zealand can be attributed to "evidence-based" medical and surgical treatments and how much can be attributed to cardiovascular risk factor reductions. METHODS AND RESULTS: A cell-based mortality model was developed and refined. This model combined (1) the published effectiveness of cardiological treatments and risk factor reductions with (2) data on all medical and surgical treatments administered to all CHD patients and (3) trends in population cardiovascular risk factors (principally smoking, cholesterol, and hypertension) from 1982 to 1993 in Auckland, New Zealand (population 996 000). Between 1982 and 1993, CHD mortality rates fell by 23.6%, with 671 fewer CHD deaths than expected from baseline mortality rates in 1982. Forty-six percent of this fall was attributed to treatments (acute myocardial infarction 12%, secondary prevention 12%, hypertension 7%, heart failure 6%, and angina 9%), and 54% was attributed to risk factor reductions (smoking 30%, cholesterol 12%, population blood pressure 8%, and other, unidentified factors 4%). These proportions remained relatively consistent after a robust sensitivity analysis. CONCLUSIONS: Approximately half the CHD mortality rate fall in Auckland, New Zealand, was attributed to medical therapies, and approximately half was attributed to reductions in major risk factors. These findings emphasize the importance of a comprehensive strategy that maximizes the population coverage of effective treatments and actively promotes a prevention program, particularly for smoking, diet, and blood pressure reduction. PMID- 11004142 TI - Differential pacing for distinguishing block from persistent conduction through an ablation line. AB - BACKGROUND: Because complete linear conduction block is necessary to minimize the recurrence of reentrant tachycardias such as typical atrial flutter, we investigated a simple technique to recognize a persistent gap or complete linear block. METHODS AND RESULTS: We prospectively evaluated cavotricuspid isthmus conduction in 50 patients (age 63+/-8 years, 43 men) after radiofrequency ablation. The distal and proximal bipoles of a quadripolar catheter placed close to the ablation line were successively stimulated during recording from the ablation line. We hypothesized that because the initial and terminal components of local potentials reflected activation at the ipsilateral and contralateral borders of the ablation lesion, a change to a more proximal pacing site without moving the catheter would prolong the stimulus to the initial component timing, whereas the response of the terminal component would depend on the presence of block or persistent conduction. A shortening or no change in timing of the terminal component would indicate block, whereas lengthening would indicate persistent gap conduction. The results were compared with previously described criteria for isthmus block. Ninety-two sites were assessed: 17 before and 75 after the achievement of complete isthmus block. The timing of the initial component was delayed by 19+/-9 ms, and the terminal component was advanced by 13+/-8 ms after block and delayed by 12+/-9 ms in case of persisting conduction. The sensitivity, specificity, and positive and negative predictive values for linear block were 100%, 75%, 94%, and 100%, respectively. CONCLUSIONS: An accurate assessment of isthmus block or persistent isthmus conduction is possible with this technique of differential pacing. PMID- 11004143 TI - Predicting outcome of defibrillation by spectral characterization and nonparametric classification of ventricular fibrillation in patients with out-of hospital cardiac arrest. AB - BACKGROUND: In 156 patients with out-of-hospital cardiac arrest of cardiac cause, we analyzed the ability of 4 spectral features of ventricular fibrillation before a total of 868 shocks to discriminate or not between segments that correspond to return of spontaneous circulation (ROSC). METHODS AND RESULTS: Centroid frequency, peak power frequency, spectral flatness, and energy were studied. A second decorrelated feature set was generated with the coefficients of the principal component analysis transformation of the original feature set. Each feature set was split into training and testing sets for improved reliability in the evaluation of nonparametric classifiers for each possible feature combination. The combination of centroid frequency and peak power frequency achieved a mean+/-SD sensitivity of 92+/-2% and specificity of 27+/-2% in testing. The highest performing classifier corresponded to the combination of the 2 dominant decorrelated spectral features with sensitivity and specificity equal to 92+/-2% and 42+/-1% in testing or a positive predictive value of 0.15 and a negative predictive value of 0.98. Using the highest performing classifier, 328 of 781 shocks not leading to ROSC would have been avoided, whereas 7 of 87 shocks leading to ROSC would not have been administered. CONCLUSIONS: The ECG contained information predictive of shock therapy. This could reduce the delivery of unsuccessful shocks and thereby the duration of unnecessary "hands-off" intervals during cardiopulmonary resuscitation. The low specificity and positive predictive value indicate that other features should be added to improve performance. PMID- 11004144 TI - Predictors of atrial fibrillation after conventional and beating heart coronary surgery: A prospective, randomized study. AB - BACKGROUND: Atrial fibrillation (AF) increases the morbidity of CABG. The pathophysiology is uncertain, and its prevention remains suboptimal. This prospective, randomized study was designed to define the role of cardiopulmonary bypass (CPB) and cardioplegic arrest in the pathogenesis of this complication. METHODS AND RESULTS: Two hundred patients were prospectively randomized to (1) on pump conventional surgery [(100 patients, 79 men, mean age 63 (40 to 77) years)] with normothermic CPB and cardioplegic arrest of the heart or (2) off-pump surgery [(100 patients, 82 men, mean age 63 (38 to 86) years)] on the beating heart. Heart rate and rhythm were continuously monitored with an automated arrhythmia detector during the first 72 hours after surgery. Thereafter, routine clinical observation was performed and continuous monitoring restarted in the case of arrhythmia. The association of perioperative factors with AF was investigated by univariate analysis. Significant variables were then included into a stepwise logistic regression model to ascertain their independent influence on the occurrence of AF. There were no significant baseline differences between groups. Thirty-nine patients in the on-pump group and 8 patients in the off-pump group had postoperative sustained AF (P:=0.001). Univariate analysis showed that CPB inclusive of cardioplegic arrest, postoperative inotropic support, intubation time, chest infection, and hospital length of stay were predictors of AF (all P:<0.05). However, stepwise multivariate regression analysis identified CPB inclusive of cardioplegic arrest as the only independent predictor of postoperative AF (OR 7.4; CI 3.4 to 17.9). CONCLUSIONS: CPB inclusive of cardioplegic arrest is the main independent predictor of postoperative AF in patients undergoing coronary revascularization. PMID- 11004145 TI - Relationship between circadian blood pressure patterns and progression of early carotid atherosclerosis: A 3-year follow-up study. AB - BACKGROUND: Arterial hypertension is a major risk factor for cardiovascular damage. The results of several studies suggest that target organ damage is greater in hypertensive persons with high blood pressure variability. METHODS AND RESULTS: During 3.3 years of follow-up, we studied the relationship between circadian blood pressure changes and the progression of early carotid atherosclerosis in 286 patients aged >55 years. Blood pressure patterns were evaluated with a long-term blood pressure monitor, and the extent of atherosclerosis was measured as the intima-media wall thickness (IMT) of the common carotid artery. Patients were subdivided according to blood pressure variability. The progression of IMT was significantly greater in the patients with increased systolic blood pressure variability (0.11 mm/y [95% CI 0.09 to 0.14] versus 0.05 mm/y [0.03 to 0.08]; P:<0.005) even after adjustment for other risk factors. Multivariate regression analysis revealed the daytime systolic blood pressure variability to be the best predictor for the progression of IMT. Raised daytime systolic blood pressure variability (>15 mm Hg) is associated with an increased relative risk of the development of early atherosclerosis (3.9 [1.4 to 11.1]; P:<0.01) and of cardiovascular events (1.87 [1.08 to 3.20]; P:<0.01). CONCLUSIONS: The daytime systolic blood pressure variability is a strong predictor of early carotid atherosclerosis progression and is useful to define the risk-benefit ratio of therapeutic approaches. PMID- 11004146 TI - Value of serum-soluble intercellular adhesion molecule-1 for the noninvasive risk assessment of transplant coronary artery disease, posttransplant ischemic events, and cardiac graft failure. AB - BACKGROUND: Adhesion molecules on arterial endothelium have been implicated in spontaneous atherosclerosis and transplant coronary artery disease (CAD). We studied whether elevated serum-soluble intercellular adhesion molecule-1 (sICAM 1) during the immediate posttransplant period was a risk factor for CAD, posttransplant ischemic events, or cardiac graft failure. METHODS AND RESULTS: We initially studied serum sICAM-1 in a subset of 16 cardiac allograft recipients (5.5+/-0.7 samples per patient) to determine a cutoff point that best correlated with presence of arterial and arteriolar endothelial ICAM-1 in matching endomyocardial biopsies. The cutoff value was 308 ng/mL. Subsequently, we prospectively evaluated serum sICAM-1 in serial samples (5.3+/-0.1 per patient) obtained during the first 3 months after transplantation in a validation subset of 130 recipients and correlated early sICAM-1 levels with long-term outcome. Serum sICAM-1 >308 ng/mL correlated significantly with ICAM-1 on arterial and arteriolar endothelium (P:=0.02). Cardiac allograft recipients with serum sICAM-1 >308 ng/mL had 2.67 (95% CI, 1.28 to 5.59, P:=0.009) times greater risk of CAD and 3.63 (95% CI, 1.05 to 12.5, P:=0.04) times greater risk of graft failure. Recipients with sICAM-1 >308 ng/mL also developed more severe CAD (P:=0.009) and more ischemic events (P:=0.03) after transplantation. CONCLUSIONS: Serum sICAM-1 levels can be used to noninvasively assess risk of transplant CAD, posttransplant ischemic events, and cardiac graft failure. PMID- 11004147 TI - Angiotensin-converting enzyme inhibition and angiotensin II type 1 receptor blockade prevent cardiac remodeling in pigs after myocardial infarction: role of tissue angiotensin II. AB - BACKGROUND: The mechanisms behind the beneficial effects of renin-angiotensin system blockade after myocardial infarction (MI) are not fully elucidated but may include interference with tissue angiotensin II (Ang II). METHODS AND RESULTS: Forty-nine pigs underwent coronary artery ligation or sham operation and were studied up to 6 weeks. To determine coronary angiotensin I (Ang I) to Ang II conversion and to distinguish plasma-derived Ang II from locally synthesized Ang II, (125)I-labeled and endogenous Ang I and II were measured in plasma and in infarcted and noninfarcted left ventricle (LV) during (125)I-Ang I infusion. Ang II type 1 (AT(1)) receptor-mediated uptake of circulating (125)I-Ang II was increased at 1 and 3 weeks in noninfarcted LV, and this uptake was the main cause of the transient elevation in Ang II levels in the noninfarcted LV at 1 week. Ang II levels and AT(1) receptor-mediated uptake of circulating Ang II were reduced in the infarct area at all time points. Coronary Ang I to Ang II conversion was unaffected by MI. Captopril and the AT(1) receptor antagonist eprosartan attenuated postinfarct remodeling, although both drugs increased cardiac Ang II production. Captopril blocked coronary conversion by >80% and normalized Ang II uptake in the noninfarcted LV. Eprosartan did not affect coronary conversion and blocked cardiac Ang II uptake by >90%. CONCLUSIONS: Both circulating and locally generated Ang II contribute to remodeling after MI. The rise in tissue Ang II production during angiotensin-converting enzyme inhibition and AT(1) receptor blockade suggests that the antihypertrophic effects of these drugs result not only from diminished AT(1) receptor stimulation but also from increased stimulation of growth-inhibitory Ang II type 2 receptors. PMID- 11004148 TI - Cardiomyocyte death induced by myocardial ischemia and reperfusion: measurement with recombinant human annexin-V in a mouse model. AB - INTRODUCTION: Phosphatidylserine (PS) externalization is regarded as one of the earliest hallmarks of cells undergoing programmed cell death. We studied the use of labeled human recombinant annexin-V, a protein selectively binding to PS, to detect cardiomyocyte death in an in vivo mouse model of cardiac ischemia and reperfusion (I/R). METHODS AND RESULTS: I/R was induced in mouse hearts by ligation and subsequent release of a suture around the left anterior descending coronary artery. Annexin-V (25 mg/kg) fused to a marker molecule was injected intra-arterially 30 minutes before euthanasia. After 15 minutes of ischemia followed by 30 minutes of reperfusion, 1.4+/-1. 2% (mean+/-SD) of the cardiomyocytes in the area at risk were annexin-V positive (n=6). This increased to 11.4+/-1.9% after 15 minutes of ischemia followed by 90 minutes of reperfusion (n=7) and to 20.2+/-3.3% after 30 minutes of ischemia followed by 90 minutes of reperfusion (n=7). In control mice, including those injected with annexin-V at the binding site of PS, no annexin-V-positive cells were observed. DNA gel electrophoresis showed typical laddering starting after 15 minutes of ischemia followed by 30 minutes of reperfusion, suggesting activation of the cell death program. Intervention in the cell death program by pretreatment with a novel Na(+)-H(+) exchange inhibitor substantially decreased annexin-V-positive cardiomyocytes from 20.2% to 2.2% in mice after 30 minutes of ischemia followed by 90 minutes of reperfusion. CONCLUSIONS: These data suggest that labeled annexin-V is useful for in situ detection of cell death in an in vivo model of I/R in the heart and for the evaluation of cell death-blocking strategies. PMID- 11004149 TI - Mechanisms of ventricular fibrillation induction by 60-Hz alternating current in isolated swine right ventricle. AB - BACKGROUND: The mechanisms by which 60-Hz alternating current (AC) can induce ventricular fibrillation (VF) are unknown. METHODS AND RESULTS: We studied 7 isolated perfused swine right ventricles in vitro. The action potential duration restitution curve was determined. Optical mapping techniques were used to determine the patterns of activation on the epicardium during 5-second 60-Hz AC stimulation (10 to 999 microA). AC captured the right ventricles at 100+/-65 microA, which is significantly lower than the direct current pacing threshold (0.77+/-0.45 mA, P:<0.05). AC induced ventricular tachycardia or VF at 477+/-266 microA, when the stimulated responses to AC had (1) short activation CLs (128+/ 14 ms), (2) short diastolic intervals (16+/-9 ms), and (3) short diastolic intervals associated with a steep action potential duration restitution curve. Optical mapping studies showed that during rapid ventricular stimulation by AC, a wave front might encounter the refractory tail of an earlier wave front, resulting in the formation of a wave break and VF. Computer simulations reproduced these results. CONCLUSIONS: AC at strengths less than the regular pacing threshold can capture the ventricle at fast rates. Accidental AC leak to the ventricles could precipitate VF and sudden death if AC results in a fast ventricular rate coupled with a steep restitution curve and a nonuniform recovery of excitability of the myocardium. PMID- 11004150 TI - Intrapericardial paclitaxel delivery inhibits neointimal proliferation and promotes arterial enlargement after porcine coronary overstretch. AB - BACKGROUND: Catheter-based intrapericardial (IPC) delivery of therapeutic agents has recently been demonstrated. Paclitaxel is known to inhibit vascular smooth muscle cell proliferation. This study examined the effect of IPC instillation of paclitaxel on neointimal proliferation after balloon overstretch of porcine coronary arteries. METHODS AND RESULTS: Overstretch injury of coronary arteries was followed by IPC administration of micellar paclitaxel at low dose (LD, 10 mg; n=6) or high dose (HD, 50 mg; n=7) or of control micelles (50 mg, n=5). Animals were euthanized 28 days after balloon dilation. Arterial injury indices were no different among the groups. The neointimal area, maximal intimal thickness, and adventitial thickness were significantly reduced in both LD (0.47+/-0.04 mm(2), 0.43+/-0.03 mm, and 0.35+/-0.02 mm, respectively) and HD (0.51+/-0.06 mm(2), 0.42+/-0.03 mm, and 0. 38+/-0.03 mm, respectively) paclitaxel groups compared with the control group (0.79+/-0.07 mm(2), 0.56+/-0.02 mm, and 0.47+/-0.02 mm, respectively; P:<0.001). Meanwhile, the vessel circumference measured at the external elastic lamina of paclitaxel-treated vessels was significantly larger than the control circumference. Apoptotic cells were found in the neointima. The apoptotic cell percentage was not different between the control (1.72%) and LD (2. 31%) groups but was higher in the HD group (7.07%, P:<0.0001 versus control and LD groups). Immunostaining for matrix metalloproteinase-2 revealed concurrent reduction in the HD group compared with the control and LD groups. CONCLUSIONS: IPC space delivery of a single dose of paclitaxel significantly reduces vessel narrowing in this balloon-overstretch model. This effect is mediated by reduction of neointimal mass as well as positive vascular remodeling. PMID- 11004151 TI - Cardiac endothelin system impairs left ventricular function in renin-dependent hypertension via decreased sarcoplasmic reticulum Ca(2+) uptake. AB - BACKGROUND: We evaluated the role of the cardiac endothelin (ET) system in compensated hypertensive left ventricular (LV) hypertrophy (LVH) and after the transition toward LV dysfunction. METHODS AND RESULTS: Hypertensive transgenic rats overexpressing the Ren2 gene (Ren2 rats) were investigated between the ages of 10 and 30 weeks (Ren2-10 and Ren2-30 groups, respectively) and compared with age-matched normotensive Sprague-Dawley (SD) rats (SD-10 and SD-30 groups, respectively). Systolic blood pressure and LV weight were elevated in both Ren2 groups compared with their age-matched SD control groups (P:<0.0001). In Ren2-30 rats, LV end-diastolic pressure increased and -dP/dt(max) decreased compared with the values in SD-30 and Ren2-10 rats (P:<0.05). This was paralleled by an activation of LV mRNA expression of preproET-1 and ET-converting enzyme-1 and ET subtype A (ETA) receptor binding in Ren2-30 compared with Ren2-10 rats (P:<0.001). Cardiac fibrosis was increased and sarcoplasmic reticulum (SR) Ca(2+) reuptake was reduced in Ren2-30 compared with SD-30 and Ren2-10 rats (P:<0.05). Treatment of Ren2 rats with the selective ETA receptor antagonist Lu135252 between 10 and 30 weeks of age did not lower systolic blood pressure, heart weight, or cardiac fibrosis but completely prevented the deterioration of LV end diastolic pressure and abolished alterations in -dP/dt(max) and SR Ca(2+) reuptake compared with no treatment in Ren2-30 and SD-30 rats (P:<0.05). CONCLUSIONS: Activation of the cardiac ET system accounts at least in part for the LV dysfunction that gradually develops in LVH. The protective effect of ETA antagonism can be attributed to the improvement of diastolic LV function that is due to normalization of impaired SR Ca(2+) uptake. PMID- 11004152 TI - Coronary artery angiography using multislice computed tomography images. PMID- 11004153 TI - Assessment of functional capacity in clinical and research applications: An advisory from the Committee on Exercise, Rehabilitation, and Prevention, Council on Clinical Cardiology, American Heart Association. PMID- 11004154 TI - Distal embolization: A threat to the coronary artery? PMID- 11004155 TI - C-reactive protein and hormone replacement therapy. PMID- 11004156 TI - Scared to death. PMID- 11004157 TI - Decreased apoptosis and tissue factor expression after lipid lowering. PMID- 11004158 TI - Dilated cardiomyopathy and the desmin gene. PMID- 11004159 TI - Mitral valve aneurysm in endocarditis. PMID- 11004160 TI - Keys to symbiotic harmony. PMID- 11004161 TI - ssgA is essential for sporulation of Streptomyces coelicolor A3(2) and affects hyphal development by stimulating septum formation. AB - The role of ssgA in cell division and development of streptomycetes was analyzed. An ssgA null mutant of Streptomyces coelicolor produced aerial hyphae but failed to sporulate, and ssgA can therefore be regarded as a novel whi gene. In addition to the morphological changes, antibiotic production was also disturbed, with strongly reduced actinorhodin production. These defects could be complemented by plasmid-borne ssgA. In the wild-type strain, transcription of ssgA was induced by nutritional shift-down and was shown to be linked to that of the upstream-located gene ssgR, which belongs to the family of iclR-type transcriptional regulator genes. Analysis of mycelium harvested from liquid-grown cultures by transmission electron microscopy showed that septum formation had strongly increased in ssgA overexpressing strains in comparison to wild-type S. coelicolor and that spore like compartments were produced at high frequency. Furthermore, the hyphae were significantly wider and contained irregular and often extremely thick septa. These data underline the important role for ssgA in Streptomyces cell division. PMID- 11004162 TI - Effects of nonpolar mutations in each of the seven Bacillus subtilis mrp genes suggest complex interactions among the gene products in support of Na(+) and alkali but not cholate resistance. AB - The Bacillus subtilis mrp (multiple resistance and pH) operon supports Na(+) and alkali resistance via an Na(+)/H(+) antiport, as well as cholate efflux and resistance. Among the individual mutants with nonpolar mutations in each of the seven mrp genes, only the mrpF mutant exhibited cholate sensitivity and a cholate efflux defect that were complemented by expression of the deleted gene in trans. Expression of mrpF in the mrp null (VKN1) strain also restored cholate transport and increased Na(+) efflux, indicating that MrpF does not require even low levels of other mrp gene expression for its own function. In contrast to MrpF, MrpA function had earlier seemed to depend upon at least modest expression of other mrp genes, i.e., mrpA restored Na(+) resistance and efflux to strain VK6 (a polar mrpA mutant which expresses low levels of mrpB to -G) but not to the null strain VKN1. In a wild-type background, each nonpolar mutation in individual mrp genes caused profound Na(+) sensitivity at both pH 7.0 and 8.3. The mrpA and mrpD mutants were particularly sensitive to alkaline pH even without added Na(+). While transport assays in membrane vesicles from selected strains indicated that MrpA-dependent antiport can occur by a secondary, proton motive force-dependent mechanism, the requirement for multiple mrp gene products suggests that there are features of energization, function, or stabilization that differ from typical secondary membrane transporters. Northern analyses indicated regulatory relationships among mrp genes as well. All the mrp mutants, especially the mrpA, B, -D, -E, and -G mutants, had elevated levels of mrp RNA relative to the wild type. Expression of an upstream gene, maeN, that encodes an Na(+)/malate symporter, was coordinately regulated with mrp, although it is not part of the operon. PMID- 11004163 TI - Covariance of complementary rRNA loop nucleotides does not necessarily represent functional pseudoknot formation in vivo. AB - We examined mutationally a two-hairpin structure (nucleotides 57 to 70 and 76 to 110) in a region of domain I of Escherichia coli 23S rRNA that has been implicated in specific functions in protein synthesis by other studies. On the basis of the observed covariance of several nucleotides in each loop in Bacteria, Archaea, and chloroplasts, the two hairpins have been proposed to form a pseudoknot. Here, appropriate loop changes were introduced in vitro by site directed mutagenesis to eliminate any possibility of base pairing between the loops. The bacterial cells containing each cloned mutant rRNA operon were then examined for cell growth, termination codon readthrough, and assembly of the mutant rRNAs into functional ribosomes. The results show that, under the conditions examined, the two hairpins do not form a pseudoknot structure that is required for the functioning of the ribosome in vivo and therefore that sequence covariance does not necessarily indicate the formation of a functional pseudoknot. PMID- 11004164 TI - Synergistic hydrolysis of carboxymethyl cellulose and acid-swollen cellulose by two endoglucanases (CelZ and CelY) from Erwinia chrysanthemi. AB - Erwinia chrysanthemi produces a battery of hydrolases and lyases which are very effective in the maceration of plant cell walls. Although two endoglucanases (CelZ and CelY; formerly EGZ and EGY) are produced, CelZ represents approximately 95% of the total carboxymethyl cellulase activity. In this study, we have examined the effectiveness of CelY and CelZ alone and of combinations of both enzymes using carboxymethyl cellulose (CMC) and amorphous cellulose (acid-swollen cellulose) as substrates. Synergy was observed with both substrates. Maximal synergy (1.8-fold) was observed for combinations containing primarily CelZ; the ratio of enzyme activities produced was similar to those produced by cultures of E. chrysanthemi. CelY and CelZ were quite different in substrate preference. CelY was unable to hydrolyze soluble cellooligosaccharides (cellotetraose and cellopentaose) but hydrolyzed CMC to fragments averaging 10.7 glucosyl units. In contrast, CelZ readily hydrolyzed cellotetraose, cellopentaose, and amorphous cellulose to produce cellobiose and cellotriose as dominant products. CelZ hydrolyzed CMC to fragments averaging 3.6 glucosyl units. In combination, CelZ and CelY hydrolyzed CMC to products averaging 2.3 glucosyl units. Synergy did not require the simultaneous presence of both enzymes. Enzymatic modification of the substrate by CelY increased the rate and extent of hydrolysis by CelZ. Full synergy was retained by the sequential hydrolysis of CMC, provided CelY was used as the first enzyme. A general mechanism is proposed to explain the synergy between these two enzymes based primarily on differences in substrate preference. PMID- 11004165 TI - Transcriptional regulation of the cpr gene cluster in ortho-chlorophenol respiring Desulfitobacterium dehalogenans. AB - To characterize the expression and possible regulation of reductive dehalogenation in halorespiring bacteria, a 11.5-kb genomic fragment containing the o-chlorophenol reductive dehalogenase-encoding cprBA genes of the gram positive bacterium Desulfitobacterium dehalogenans was subjected to detailed molecular characterization. Sequence analysis revealed the presence of eight designated genes with the order cprTKZEBACD and with the same polarity except for cprT. The deduced cprC and cprK gene products belong to the NirI/NosR and CRP-FNR families of transcription regulatory proteins, respectively. CprD and CprE are predicted to be molecular chaperones of the GroEL type, whereas cprT may encode a homologue of the trigger factor folding catalysts. Northern blot analysis, reverse transcriptase PCR, and primer extension analysis were used to elucidate the transcriptional organization and regulation of the cpr gene cluster. Results indicated halorespiration-specific transcriptional induction of the monocistronic cprT gene and the biscistronic cprBA and cprZE genes. Occasional read-through at cprC gives rise to a tetracistronic cprBACD transcript. Transcription of cprBA was induced 15-fold upon addition of the o-chlorophenolic substrate 3-chloro-4 hydroxyphenylacetic acid within 30 min with concomitant induction of dehalogenation activity. Putative regulatory protein binding motifs that to some extent resemble the FNR box were identified in the cprT-cprK and cprK-cprZ intergenic regions and the promoter at cprB, suggesting a role for FNR-like CprK in the control of expression of the cprTKZEBACD genes. PMID- 11004166 TI - Isolation of regulated genes of the cyanobacterium Synechocystis sp. strain PCC 6803 by differential display. AB - Global identification of differentially regulated genes in prokaryotes is constrained because the mRNA does not have a 3' polyadenylation extension; this precludes specific separation of mRNA from rRNA and tRNA and synthesis of cDNAs from the entire mRNA population. Knowledge of the entire genome sequence of Synechocystis sp. strain PCC 6803 has enabled us to develop a differential display procedure that takes advantage of a short palindromic sequence that is dispersed throughout the Synechocystis sp. strain PCC 6803 genome. This sequence, designated the HIP (highly iterated palindrome) element, occurs in approximately half of the Synechocystis sp. strain PCC 6803 genes but is absent in rRNA and tRNA genes. To determine the feasibility of exploiting the HIP element, alone or in combination with specific primer subsets, for analyzing differential gene expression, we used HIP-based primers to identify light intensity-regulated genes. Several gene fragments, including those encoding ribosomal proteins and phycobiliprotein subunits, were differentially amplified from RNA templates derived from cells grown in low light or exposed to high light for 3 h. One novel finding was that expression of certain genes of the pho regulon, which are under the control of environmental phosphate levels, were markedly elevated in high light. High-light activation of pho regulon genes correlated with elevated growth rates that occur when the cells are transferred from low to high light. These results suggest that in high light, the rate of growth of Synechocystis sp. strain PCC 6803 exceeds its capacity to assimilate phosphate, which, in turn, may trigger a phosphate starvation response and activation of the pho regulon. PMID- 11004167 TI - The LE1 bacteriophage replicates as a plasmid within Leptospira biflexa: construction of an L. biflexa-Escherichia coli shuttle vector. AB - We have discovered that LE1, one of the plaque-forming phages previously described as lytic for the Leptospira biflexa saprophytic spirochete (I. Saint Girons, D. Margarita, P. Amouriaux, and G. Baranton, Res. Microbiol. 141:1131 1138, 1990), was indeed temperate. LE1 was found to be unusual, as Southern blot analysis indicated that it is one of the few phages to replicate in the prophage state as a circular plasmid. The unavailability of such small endogenous replicons has hindered genetic experimentation in Leptospira. We have developed a shuttle vector with DNA derived from LE1. Random LE1 DNA fragments were cloned into a pGEM 7Zf(+) derivative devoid of most of the bla gene but carrying a kanamycin resistance marker from the gram-positive bacterium Enterococcus (Streptococcus) faecalis. These constructs were transformed into L. biflexa strain Patoc 1 by electroporation, giving rise to kanamycin-resistant transformants. A 2.2-kb fragment from LE1 was responsible for replication of the vector in L. biflexa. However, a larger region including an intact parA gene homologue was necessary for the stability of the shuttle vector. Direct repeats and AT-rich regions characterized the LE1 origin of replication. Our data indicate that the replicon derived from the LE1 leptophage, together with the kanamycin resistance gene, is a promising tool with which to develop the genetics of Leptospira species. PMID- 11004168 TI - Role of the Escherichia coli nucleotide excision repair proteins in DNA replication. AB - DNA polymerase I (PolI) functions both in nucleotide excision repair (NER) and in the processing of Okazaki fragments that are generated on the lagging strand during DNA replication. Escherichia coli cells completely lacking the PolI enzyme are viable as long as they are grown on minimal medium. Here we show that viability is fully dependent on the presence of functional UvrA, UvrB, and UvrD (helicase II) proteins but does not require UvrC. In contrast, delta polA cells grow even better when the uvrC gene has been deleted. Apparently UvrA, UvrB, and UvrD are needed in a replication backup system that replaces the PolI function, and UvrC interferes with this alternative replication pathway. With specific mutants of UvrC we could show that the inhibitory effect of this protein is related to its catalytic activity that on damaged DNA is responsible for the 3' incision reaction. Specific mutants of UvrA and UvrB were also studied for their capacity to support the PolI-independent replication. Deletion of the UvrC binding domain of UvrB resulted in a phenotype similar to that caused by deletion of the uvrC gene, showing that the inhibitory incision activity of UvrC is mediated via binding to UvrB. A mutation in the N-terminal zinc finger domain of UvrA does not affect NER in vivo or in vitro. The same mutation, however, does give inviability in combination with the delta polA mutation. Apparently the N terminal zinc-binding domain of UvrA has specifically evolved for a function outside DNA repair. A model for the function of the UvrA, UvrB, and UvrD proteins in the alternative replication pathway is discussed. PMID- 11004169 TI - glnE is an essential gene in Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis possesses a homologue of glnE, potentially encoding a regulator of glutamine synthetase activity. We attempted to construct glnE disrupted mutants using a two-step strategy, whereby a single-crossover strain was first isolated, followed by sacB counterselection to isolate the double crossover strain. Of 192 sucrose-resistant colonies tested, none were mutants, although the wild-type double crossover could be easily isolated. When a second copy of the wild-type glnE was integrated into the chromosome, we could isolate both wild-type and mutant double-crossover strains. Thus, the chromosomal gene could only be replaced with a disrupted copy when another functional copy of the gene was provided, demonstrating that this gene is essential under the conditions tested. PMID- 11004170 TI - Population studies of methicillin-resistant and -sensitive Staphylococcus aureus strains reveal a lack of variability in the agrD gene, encoding a staphylococcal autoinducer peptide. AB - The virulence of Staphylococcus aureus is controlled by the accessory gene regulator (agr) system, including an extracellular inducer encoded by agrD. Variable agr PCR restriction fragment length polymorphism (RFLP) patterns of unique S. aureus strains (n = 192) were determined for a region comprising agrD and parts of the neighboring agrC and agrB genes. Twelve unique RFLP patterns were identified among S. aureus strains in general; these patterns were further specified by sequencing. All sequences could be catalogued in the three current agr groups. A major proportion of the S. aureus strains belong to agr group 1, whereas only 6% of the methicillin-susceptible S. aureus strains and 5% of the methicillin-resistant S. aureus strains belong to agr groups 2 and 3, respectively. The homology between groups varied from 75 to 80%, and within groups it varied from 96 to 100%. Different levels of sequence variability were observed in the different agr genes. agr-related bacterial interference among colonizing S. aureus strains in the noses of persistent and intermittent human carriers was studied. S. aureus strains belonging to different agr groups were encountered in the same individual. This may suggest that the activity of the agrD gene product does not define colonization dynamics, which is further substantiated by the rarity of agr group 2 and 3 strains. PMID- 11004171 TI - A single-transformation gene function test in diploid Candida albicans. AB - The fungal pathogen Candida albicans is naturally diploid, and current gene disruption strategies require two successive transformations. We describe here a genetic construct (UAU1) for which two copies may be selected. Insertion of UAU1 into one genomic site, after a single transformation, allows selection for segregants with two copies of the insertion. Major classes of segregants are those carrying homozygous insertion mutations and allelic triplications, which have two insertion alleles and a wild-type allele. Thus nonessential and essential genes may be distinguished rapidly through PCR tests for homozygosis and triplication. We find that homozygous mutations may be isolated at three nonessential loci (ADE2, RIM20, and YGR189), while only allelic triplications were found at two essential loci (SNF1 and CDC28). We have unexpectedly isolated homozygous mutants with mutations at CDC25; they are viable but defective in filamentation on serum-containing medium. The UAU1 cassette is thus useful to assess rapidly the essentiality of C. albicans genes. PMID- 11004172 TI - Characterization and expression of HmuR, a TonB-dependent hemoglobin receptor of Porphyromonas gingivalis. AB - The gram-negative pathogen Porphyromonas gingivalis requires hemin for growth. Hemoglobin bound to haptoglobin and hemin complexed to hemopexin can be used as heme sources, indicating that P. gingivalis must have a means to remove the hemin from these host iron-binding proteins. However, the specific mechanisms utilized by P. gingivalis for the extraction of heme from heme-binding proteins and for iron transport are poorly understood. In this study we have determined that a newly identified TonB-dependent hemoglobin-hemin receptor (HmuR) is involved in hemoglobin binding and utilization in P. gingivalis A7436. HmuR shares amino acid homology with TonB-dependent outer membrane receptors of gram-negative bacteria involved in the acquisition of iron from hemin and hemoglobin, including HemR of Yersinia enterocolitica, ShuA of Shigella dysenteriae, HpuB of Neisseria gonorrhoeae and N. meningitidis, HmbR of N. meningitidis, HgbA of Haemophilus ducreyi, and HgpB of H. influenzae. Southern blot analysis confirmed the presence of the hmuR gene and revealed genetic variability in the carboxy terminus of hmuR in P. gingivalis strains 33277, 381, W50, and 53977. We also identified directly upstream of the hmuR gene a gene which we designated hmuY. Upstream of the hmuY start codon, a region with homology to the Fur binding consensus sequence was identified. Reverse transcription-PCR analysis revealed that hmuR and hmuY were cotranscribed and that transcription was negatively regulated by iron. Inactivation of hmuR resulted in a decreased ability of P. gingivalis to bind hemoglobin and to grow with hemoglobin or hemin as sole iron sources. Escherichia coli cells expressing recombinant HmuR were shown to bind hemoglobin and hemin. Furthermore, purified recombinant HmuR was demonstrated to bind hemoglobin. Taken together, these results indicate that HmuR serves as the major TonB-dependent outer membrane receptor involved in the utilization of both hemin and hemoglobin in P. gingivalis. PMID- 11004173 TI - Identification of RpoS (sigma(S))-regulated genes in Salmonella enterica serovar typhimurium. AB - The rpoS gene encodes the alternative sigma factor sigma(S) (RpoS) and is required for survival of bacteria under starvation and stress conditions. It is also essential for Salmonella virulence in mice. Most work on the RpoS regulon has been in the closely related enterobacterial species Escherichia coli. To characterize the RpoS regulon in Salmonella, we isolated 38 unique RpoS-activated lacZ gene fusions from a bank of Salmonella enterica serovar Typhimurium mutants harboring random Tn5B21 mutations. Dependence on RpoS varied from 3-fold to over 95-fold, and all gene fusions isolated were regulated by growth phase. The identities of 21 RpoS-dependent fusions were determined by DNA sequence analysis. Seven of the fusions mapped to DNA regions in Salmonella serovar Typhimurium that do not match any known E. coli sequence, suggesting that the composition of the RpoS regulon differs markedly in the two species. The other 14 fusions mapped to 13 DNA regions very similar to E. coli sequences. None of the insertion mutations in DNA regions common to both species appeared to affect Salmonella virulence in BALB/c mice. Of these, only three (otsA, katE, and poxB) are located in known members of the RpoS regulon. Ten insertions mapped in nine open reading frames of unknown function (yciF, yehY, yhjY, yncC, yjgB, yahO, ygaU, ycgB, and yeaG) appear to be novel members of the RpoS regulon. One insertion, that in mutant C52::H87, was in the noncoding region upstream from ogt, encoding a O(6) methylguanine DNA methyltransferase involved in repairing alkylation damage in DNA. The ogt coding sequence is very similar to the E. coli homolog, but the ogt 5' flanking regions were found to be markedly different in the two species, suggesting genetic rearrangements. Using primer extension assays, a specific ogt mRNA start site was detected in RNAs of the Salmonella serovar Typhimurium wild type strains C52 and SL1344 but not in RNAs of the mutant strains C52K (rpoS), SL1344K (rpoS), and C52::H87. In mutant C52::H87, Tn5B21 is inserted at the ogt mRNA start site, with lacZ presumably transcribed from the identified RpoS regulated promoter. These results indicate that ogt gene expression in Salmonella is regulated by RpoS in stationary phase of growth in rich medium, a finding that suggests a novel role for RpoS in DNA repair functions. PMID- 11004175 TI - A truncated soluble Bacillus signal peptidase produced in Escherichia coli is subject to self-cleavage at its active site. AB - Soluble forms of Bacillus signal peptidases which lack their unique amino terminal membrane anchor are prone to degradation, which precludes their high level production in the cytoplasm of Escherichia coli. Here, we show that the degradation of soluble forms of the Bacillus signal peptidase SipS is largely due to self-cleavage. First, catalytically inactive soluble forms of this signal peptidase were not prone to degradation; in fact, these mutant proteins were produced at very high levels in E. coli. Second, the purified active soluble form of SipS displayed self-cleavage in vitro. Third, as determined by N-terminal sequencing, at least one of the sites of self-cleavage (between Ser15 and Met16 of the truncated enzyme) strongly resembles a typical signal peptidase cleavage site. Self-cleavage at the latter position results in complete inactivation of the enzyme, as Ser15 forms a catalytic dyad with Lys55. Ironically, self-cleavage between Ser15 and Met16 cannot be prevented by mutagenesis of Gly13 and Ser15, which conform to the -1, -3 rule for signal peptidase recognition, because these residues are critical for signal peptidase activity. PMID- 11004174 TI - Transport of C(4)-dicarboxylates in Wolinella succinogenes. AB - C(4)-dicarboxylate transport is a prerequisite for anaerobic respiration with fumarate in Wolinella succinogenes, since the substrate site of fumarate reductase is oriented towards the cytoplasmic side of the membrane. W. succinogenes was found to transport C(4)-dicarboxylates (fumarate, succinate, malate, and aspartate) across the cytoplasmic membrane by antiport and uniport mechanisms. The electrogenic uniport resulted in dicarboxylate accumulation driven by anaerobic respiration. The molar ratio of internal to external dicarboxylate concentration was up to 10(3). The dicarboxylate antiport was either electrogenic or electroneutral. The electroneutral antiport required the presence of internal Na(+), whereas the electrogenic antiport also operated in the absence of Na(+). In the absence of Na(+), no electrochemical proton potential (delta p) was measured across the membrane of cells catalyzing fumarate respiration. This suggests that the proton potential generated by fumarate respiration is dissipated by the concomitant electrogenic dicarboxylate antiport. Three gene loci (dcuA, dcuB, and dctPQM) encoding putative C(4)-dicarboxylate transporters were identified on the genome of W. succinogenes. The predicted gene products of dcuA and dcuB are similar to the Dcu transporters that are involved in the fumarate respiration of Escherichia coli with external C(4) dicarboxylates. The genes dctP, -Q, and -M probably encode a binding-protein dependent secondary uptake transporter for dicarboxylates. A mutant (DcuA(-) DcuB(-)) of W. succinogenes lacking the intact dcuA and dcuB genes grew by nitrate respiration with succinate as the carbon source but did not grow by fumarate respiration with fumarate, malate, or aspartate as substrates. The DcuA( ), DcuB(-), and DctQM(-) mutants grew by fumarate respiration as well as by nitrate respiration with succinate as the carbon source. Cells of the DcuA(-) DcuB(-) mutant performed fumarate respiration without generating a proton potential even in the presence of Na(+). This explains why the DcuA(-) DcuB(-) mutant does not grow by fumarate respiration. Growth by fumarate respiration appears to depend on the function of the Na(+)-dependent, electroneutral dicarboxylate antiport which is catalyzed exclusively by the Dcu transporters. Dicarboxylate transport via the electrogenic uniport is probably catalyzed by the DctPQM transporter and by a fourth, unknown transporter that may also operate as an electrogenic antiporter. PMID- 11004176 TI - Polyprenyl phosphate biosynthesis in Mycobacterium tuberculosis and Mycobacterium smegmatis. AB - Mycobacterium smegmatis has been shown to contain two forms of polyprenyl phosphate (Pol-P), while Mycobacterium tuberculosis contains only one. Utilizing subcellular fractions from M. smegmatis and M. tuberculosis, we show that Pol-P synthesis is different in these species. The specific activities of the prenyl diphosphate synthases in M. tuberculosis are 10- to 100-fold lower than those in M. smegmatis. In M. smegmatis decaprenyl diphosphate and heptaprenyl diphosphate were the main products synthesized in vitro, whereas in M. tuberculosis only decaprenyl diphosphate was synthesized. The data from both organisms suggest that geranyl diphosphate is the allylic substrate for two distinct prenyl diphosphate synthases, one located in the cell membrane that synthesizes omega,E,Z-farnesyl diphosphate and the other present in the cytosol that synthesizes omega,E,E,E geranylgeranyl diphosphate. In M. smegmatis, the omega,E, Z-farnesyl diphosphate is utilized by a membrane-associated prenyl diphosphate synthase activity to generate decaprenyl diphosphate, and the omega,E,E,E-geranylgeranyl diphosphate is utilized by a membrane-associated activity for the synthesis of the heptaprenyl diphosphate. In M. tuberculosis, however, omega,E,E,E-geranylgeranyl diphosphate is not utilized for the synthesis of heptaprenyl diphosphate. Thus, the difference in the compositions of the Pol-P of M. smegmatis and M. tuberculosis can be attributed to distinct enzymatic differences between these two organisms. PMID- 11004177 TI - The torYZ (yecK bisZ) operon encodes a third respiratory trimethylamine N-oxide reductase in Escherichia coli. AB - The bisZ gene of Escherichia coli was previously described as encoding a minor biotin sulfoxide (BSO) reductase in addition to the main cytoplasmic BSO reductase, BisC. In this study, bisZ has been renamed torZ based on the findings that (i) the torZ gene product, TorZ, is able to reduce trimethylamine N-oxide (TMAO) more efficiently than BSO; (ii) although TorZ is more homologous to BisC than to the TMAO reductase TorA (63 and 42% identity, respectively), it is located mainly in the periplasm as is TorA; (iii) torZ belongs to the torYZ operon, and the first gene, torY (formerly yecK), encodes a pentahemic c-type cytochrome homologous to the TorC cytochrome of the TorCAD respiratory system. Furthermore, the torYZ operon encodes a third TMAO respiratory system, with catalytic properties that are clearly different from those of the TorCAD and the DmsABC systems. The torYZ and the torCAD operons may have diverged from a common ancestor, but, surprisingly, no torD homologue is found in the sequences around torYZ. Moreover, the torYZ operon is expressed at very low levels under the conditions tested, and, in contrast to torCAD, it is not induced by TMAO or dimethyl sulfoxide. PMID- 11004178 TI - sigma(54) Promoters control expression of genes encoding the hook and basal body complex in Rhodobacter sphaeroides. AB - Gene expression of the flagellar system is tightly controlled by external stimuli or intracellular signals. A general picture of this regulation has been obtained from studies of Salmonella enterica serovar Typhimurium. However, these regulatory mechanisms do not apply to all bacterial groups. In this study, we have investigated regulation of the flagellar genetic system in Rhodobacter sphaeroides. Deletion analysis, site-directed mutagenesis, and 5'-end mapping were conducted in order to identify the fliO promoter. Our results indicate that this promoter is recognized by the factor sigma(54). Additionally, 5'-end mapping of the flgB and fliK transcripts suggests that these mRNAs are also transcribed from sigma(54) promoters. Finally, we showed evidence that suggests that fliC transcription is not entirely dependent on the presence of a complete basal body hook structure. Our results are discussed in the context of a possible regulatory hierarchy controlling flagellar gene expression in R. sphaeroides. PMID- 11004179 TI - Myxococcus xanthus dif genes are required for biogenesis of cell surface fibrils essential for social gliding motility. AB - Myxococcus xanthus social (S) gliding motility has been previously reported by us to require the chemotaxis homologues encoded by the dif genes. In addition, two cell surface structures, type IV pili and extracellular matrix fibrils, are also critical to M. xanthus S motility. We have demonstrated here that M. xanthus dif genes are required for the biogenesis of fibrils but not for that of type IV pili. Furthermore, the developmental defects of dif mutants can be partially rescued by the addition of isolated fibril materials. Along with the chemotaxis genes of various swarming bacteria and the pilGHIJ genes of the twitching bacterium Pseudomonas aeruginosa, the M. xanthus dif genes belong to a unique class of bacterial chemotaxis genes or homologues implicated in the biogenesis of structures required for bacterial surface locomotion. Genetic studies indicate that the dif genes are linked to the M. xanthus dsp region, a locus known to be crucial for M. xanthus fibril biogenesis and S gliding. PMID- 11004180 TI - Characterization of the ccpA gene of Enterococcus faecalis: identification of starvation-inducible proteins regulated by ccpA. AB - Inactivation of ccpA in Enterococcus faecalis leads to reduction of the growth rate, derepression of the galKETR operon in the presence of a mixture of glucose and galactose, and reduction of transcription of ldh in the presence of glucose. Moreover, the E. faecalis ccpA gene fully complements a Bacillus subtilis ccpA mutant, arguing for similar functions of these two homologous proteins. Protein comparison on two-dimensional gels from the wild-type cells and the ccpA mutant cells revealed a pleiotropic effect of the mutation on gene expression. The HPr protein of the carbohydrate-phosphotransferase system was identified by microsequencing, and a modification of its phosphorylation state was observed between the wild-type and the mutant strains. Moreover, at least 16 polypeptides are overexpressed in the mutant, and 6 are repressed. Interestingly, 13 of the 16 polypeptides whose synthesis is enhanced in the mutant were also identified as glucose starvation proteins. The N-terminal amino acid sequences of four of them match sequences deduced from genes coding for L-serine dehydratase, dihydroxyacetone kinase (two genes), and a protein of unknown function from Deinococcus radiodurans. PMID- 11004181 TI - Characterization of bacteriophage lambda excisionase mutants defective in DNA binding. AB - The bacteriophage lambda excisionase (Xis) is a sequence-specific DNA binding protein required for excisive recombination. Xis binds cooperatively to two DNA sites arranged as direct repeats on the phage DNA. Efficient excision is achieved through a cooperative interaction between Xis and the host-encoded factor for inversion stimulation as well as a cooperative interaction between Xis and integrase. The secondary structure of the Xis protein was predicted to contain a typical amphipathic helix that spans residues 18 to 28. Several mutants, defective in promoting excision in vivo, were isolated with mutations at positions encoding polar amino acids in the putative helix (T. E. Numrych, R. I. Gumport, and J. F. Gardner, EMBO J. 11:3797-3806, 1992). We substituted alanines for the polar amino acids in this region. Mutant proteins with substitutions for polar amino acids in the amino-terminal region of the putative helix exhibited decreased excision in vivo and were defective in DNA binding. In addition, an alanine substitution at glutamic acid 40 also resulted in altered DNA binding. This indicates that the hydrophilic face of the alpha-helix and the region containing glutamic acid 40 may form the DNA binding surfaces of the Xis protein. PMID- 11004182 TI - The nrfA and nirB nitrite reductase operons in Escherichia coli are expressed differently in response to nitrate than to nitrite. AB - Escherichia coli possesses two distinct nitrite reductase enzymes encoded by the nrfA and nirB operons. The expression of each operon is induced during anaerobic cell growth conditions and is further modulated by the presence of either nitrite or nitrate in the cells' environment. To examine how each operon is expressed at low, intermediate, and high levels of either nitrate or nitrite, anaerobic chemostat culture techniques were employed using nrfA-lacZ and nirB-lacZ reporter fusions. Steady-state gene expression studies revealed a differential pattern of nitrite reductase gene expression where optimal nrfA-lacZ expression occurred only at low to intermediate levels of nitrate and where nirB-lacZ expression was induced only by high nitrate conditions. Under these conditions, the presence of high levels of nitrate suppressed nrfA gene expression. While either NarL or NarP was able to induce nrfA-lacZ expression in response to low levels of nitrate, only NarL could repress at high nitrate levels. The different expression profile for the alternative nitrite reductase operon encoded by nirBDC under high-nitrate conditions was due to transcriptional activation by either NarL or NarP. Neither response regulator could repress nirB expression. Nitrite was also an inducer of nirB and nrfA gene expression, but nitrate was always the more potent inducer by >100-fold. Lastly, since nrfA operon expression is only induced under low-nitrate concentrations, the NrfA enzyme is predicted to have a physiological role only where nitrate (or nitrite) is limiting in the cell environment. In contrast, the nirB nitrite reductase is optimally synthesized only when nitrate or nitrite is in excess of the cell's capacity to consume it. Revised regulatory schemes are presented for NarL and NarP in control of the two operons. PMID- 11004183 TI - The N-terminal region of the Oenococcus oeni bacteriophage fOg44 lysin behaves as a bona fide signal peptide in Escherichia coli and as a cis-inhibitory element, preventing lytic activity on oenococcal cells. AB - The function of the N-terminal region of the Oenococcus oeni phage fOg44 lysin (Lys44) as an export signal was investigated. We observed that when induced in Escherichia coli, Lys44 was cleaved between residues 27 and 28 in a SecA dependent manner. Lys44 processing could be blocked by a specific signal peptidase inhibitor and was severely reduced by modification of the cleavage site. The lethal effect of Lys44 expression observed in E. coli was ascribed to the presence of its N-terminal 27-residue sequence, as its deletion resulted in the production of a nontoxic, albeit active, product. We have further established that lytic activity in oenococcal cells was dependent on Lys44 processing. An active protein with the molecular mass expected for the cleaved enzyme was detected in extracts from O. oeni-infected cells. The temporal pattern of its appearance suggests that synthesis and export of Lys44 in the infected host progress along with phage maturation. Overall, these results provide, for the first time, experimental evidence for the presence of a signal peptide in a bacteriophage lysin. Database searches and alignment of protein sequences support the prediction that other known O. oeni and Lactococcus lactis phages also encode secretory lysins. The evolutionary significance of a putative phage lysis mechanism relying on secretory lytic enzymes is tentatively discussed, on the basis of host cell wall structure and autolytic capacity. PMID- 11004184 TI - Leafy gall formation is controlled by fasR, an AraC-type regulatory gene in Rhodococcus fascians. AB - Rhodococcus fascians can interact with many plant species and induce the formation of either leafy galls or fasciations. To provoke symptoms, R. fascians strain D188 requires pathogenicity genes that are located on a linear plasmid, pFiD188. The fas genes are essential for virulence and constitute an operon that encodes, among other functions, a cytokinin synthase gene. Expression of the fas genes is induced by extracts of infected plant tissue only. We have isolated an AraC-type regulatory gene, fasR, located on pFiD188, which is indispensable for pathogenesis and for fas gene expression. The combined results of our experiments show that in vitro expression of the fas genes in a defined medium is strictly regulated and that several environmental factors (pH, carbon and nitrogen sources, phosphate and oxygen content, and cell density) and regulatory proteins are involved. We further show that expression of the fas genes is controlled at both the transcriptional and the translational levels. The complex expression pattern probably reflects the necessity of integrating a multitude of signals and underlines the importance of the fas operon in the pathogenicity of R. fascians. PMID- 11004185 TI - Evidence of a role for LytB in the nonmevalonate pathway of isoprenoid biosynthesis. AB - It is proposed that the lytB gene encodes an enzyme of the deoxyxylulose-5 phosphate (DOXP) pathway that catalyzes a step at or subsequent to the point at which the pathway branches to form isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). A mutant of the cyanobacterium Synechocystis strain PCC 6803 with an insertion in the promoter region of lytB grew slowly and produced greenish-yellow, easily bleached colonies. Insertions in the coding region of lytB were lethal. Supplementation of the culture medium with the alcohol analogues of IPP and DMAPP (3-methyl-3-buten-1-ol and 3-methyl-2-buten-1 ol) completely alleviated the growth impairment of the mutant. The Synechocystis lytB gene and a lytB cDNA from the flowering plant Adonis aestivalis were each found to significantly enhance accumulation of carotenoids in Escherichia coli engineered to produce these colored isoprenoid compounds. When combined with a cDNA encoding deoxyxylulose-5-phosphate synthase (dxs), the initial enzyme of the DOXP pathway, the individual salutary effects of lytB and dxs were multiplied. In contrast, the combination of lytB and a cDNA encoding IPP isomerase (ipi) was no more effective in enhancing carotenoid accumulation than ipi alone, indicating that the ratio of IPP and DMAPP produced via the DOXP pathway is influenced by LytB. PMID- 11004186 TI - Genes involved in anaerobic metabolism of phenol in the bacterium Thauera aromatica. AB - Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium Thauera aromatica have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor ("phenylphosphate synthase") and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate ("phenylphosphate carboxylase"). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the ubiD gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the ubiX gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross hybridized with DNA from different Thauera and Azoarcus strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the mutT gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of Pseudomonas putida. DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator. PMID- 11004187 TI - Identification of a copper-responsive two-component system on the chromosome of Escherichia coli K-12. AB - Using a genetic screen we have identified two chromosomal genes, cusRS (ylcA ybcZ), from Escherichia coli K-12 that encode a two-component, signal transduction system that is responsive to copper ions. This regulatory system is required for copper-induced expression of pcoE, a plasmid-borne gene from the E. coli copper resistance operon pco. The closest homologs of CusR and CusS are plasmid-borne two-component systems that are also involved in metal responsive gene regulation: PcoR and PcoS from the pco operon of E. coli; CopR and CopS from the cop operon, which provides copper resistance to Pseudomonas syringae; and SilR and SilS from the sil locus, which provides silver ion resistance to Salmonella enterica serovar Typhimurium. The genes cusRS are also required for the copper-dependent expression of at least one chromosomal gene, designated cusC (ylcB), which is allelic to the recently identified virulence gene ibeB in E. coli K1. The cus locus may comprise a copper ion efflux system, because the expression of cusC is induced by high concentrations of copper ions. Furthermore, the translation products of cusC and additional downstream genes are homologous to known metal ion antiporters. PMID- 11004188 TI - The last RNA-binding repeat of the Escherichia coli ribosomal protein S1 is specifically involved in autogenous control. AB - The ssyF29 mutation, originally selected as an extragenic suppressor of a protein export defect, has been mapped within the rpsA gene encoding ribosomal protein S1. Here, we examine the nature of this mutation and its effect on translation. Sequencing of the rpsA gene from the ssyF mutant has revealed that, due to an IS10R insertion, its product lacks the last 92 residues of the wild-type S1 protein corresponding to one of the four homologous repeats of the RNA-binding domain. To investigate how this truncation affects translation, we have created two series of Escherichia coli strains (rpsA(+) and ssyF) bearing various translation initiation regions (TIRs) fused to the chromosomal lacZ gene. Using a beta-galactosidase assay, we show that none of these TIRs differ in activity between ssyF and rpsA(+) cells, except for the rpsA TIR: the latter is stimulated threefold in ssyF cells, provided it retains at least ca. 90 nucleotides upstream of the start codon. Similarly, the activity of this TIR can be severely repressed in trans by excess S1, again provided it retains the same minimal upstream sequence. Thus, the ssyF stimulation requires the presence of the rpsA translational autogenous operator. As an interpretation, we propose that the ssyF mutation relieves the residual repression caused by normal supply of S1 (i.e., that it impairs autogenous control). Thus, the C-terminal repeat of the S1 RNA binding domain appears to be required for autoregulation, but not for overall mRNA recognition. PMID- 11004189 TI - Characterization of tpp1(+) as encoding a main trehalose-6P phosphatase in the fission yeast Schizosaccharomyces pombe. AB - We have characterized an open reading frame of 2,454 bp on chromosome I of Schizosaccharomyces pombe as the gene encoding trehalose-6P phosphatase (tpp1(+)). Disruption of tpp1(+) caused in vivo accumulation of trehalose-6P upon heat shock and prevented cell growth at 37 to 40 degrees C. Accumulation of trehalose-6P in cells bearing a chromosomal disruption of the tpp1(+) gene and containing a plasmid with tpp1(+) under the control of the thiamine-repressible promotor correlated with tpp1(+) repression. The level of tpp1(+) mRNA rose upon heat shock, osmostress, or oxidative stress and was negatively controlled by cyclic AMP-dependent protein kinase activity. Expression of tpp1(+) during oxidative or osmotic stress, but not during heat shock, was under positive control by the wis1-sty1 (equivalent to phh1 and spc1) mitogen-activated protein kinase pathway. Analysis of Tpp1 protein levels suggests that the synthesis of trehalose-6P phosphatase may also be subjected to translational or posttranslational control. PMID- 11004190 TI - Transcriptional control of the sulfur-regulated cysH operon, containing genes involved in L-cysteine biosynthesis in Bacillus subtilis. AB - The molecular mechanisms of regulation of the genes involved in the biosynthesis of cysteine are poorly characterized in Bacillus subtilis and other gram-positive bacteria. In this study we describe the expression pattern of the B. subtilis cysH operon in response to sulfur starvation. A 6.1-kb polycistronic transcript which includes the cysH, cysP, ylnB, ylnC, ylnD, ylnE, and ylnF genes was identified. Its synthesis was induced by sulfur limitation and strongly repressed by cysteine. The cysH operon contains a 5' leader portion homologous to that of the S box family of genes involved in sulfur metabolism, which are regulated by a transcription termination control system. Here we show that induction of B. subtilis cysH operon expression is dependent on the promoter and independent of the leader region terminator, indicating that the operon is regulated at the level of transcription initiation rather than controlled at the level of premature termination of transcription. Deletion of a 46-bp region adjacent to the -35 region of the cysH promoter led to high-level expression of the operon, even in the presence of cysteine. We also found that O-acetyl-L-serine (OAS), a direct precursor of cysteine, renders cysH transcription independent of sulfur starvation and insensitive to cysteine repression. We propose that transcription of the cysH operon is negatively regulated by a transcriptional repressor whose activity is controlled by the intracellular levels of OAS. Cysteine is predicted to repress transcription by inhibiting the synthesis of OAS, which would act as an inducer of cysH expression. These novel results provide the first direct evidence that cysteine biosynthesis is controlled at a transcriptional level by both negative and positive effectors in a gram-positive organism. PMID- 11004191 TI - SarA represses agr operon expression in a purified in vitro Staphylococcus aureus transcription system. AB - Mutation and genetic complementation studies suggested that two chromosomal loci, agr and sar, are involved in the upregulation of several exotoxin genes and the downregulation of a number of surface protein genes in a growth phase-dependent manner in Staphylococcus aureus. We purified recombinant T7-tagged SarA from Escherichia coli and determined its effect on transcription from several S. aureus promoters by using purified RNA polymerase reconstituted with either sigma(A) or sigma(B) from S. aureus. Of the seven sigma(A)-dependent promoters that we tested, SarA repressed transcription from agrP2, agrP3, cna, sarP1, and sea promoters and did not affect sec and znt promoters. Furthermore, SarA had no effect on transcription from the sigma(B)-dependent sarP3 promoter. In vitro experimental data presented in this report suggest that SarA expression is autoregulated. PMID- 11004192 TI - Null mutation of the dam or seqA gene suppresses temperature-sensitive lethality but not hypersensitivity to novobiocin of muk null mutants. AB - Escherichia coli mukF, mukE, and mukB null mutants have common phenotypes such as temperature-dependent colony formation, anucleate cell production, chromosome cutting by septum closure, and abnormal localization of SeqA-DNA clusters. We show here that the associated muk null mutations cause hypersensitivity to novobiocin. Null mutation of either dam or seqA suppressed partially the temperature-sensitive lethality but failed to suppress the anucleate cell production and the hypersensitivity to novobiocin caused by muk null mutations. PMID- 11004193 TI - New virulence-activated and virulence-repressed genes identified by systematic gene inactivation and generation of transcriptional fusions in Bordetella pertussis. AB - An in silico scan of the partially completed genome sequence of Bordetella pertussis and analyses of transcriptional fusions generated with a new integrational vector were used to identify new potential virulence genes. The genes encoding a putative siderophore receptor, adhesins, and an autotransporter protein appeared to be regulated in a manner similar to Bordetella virulence genes by the global virulence regulator BvgAS. In contrast, the gene encoding a putative intimin-like protein appeared to be repressed under conditions of virulence. PMID- 11004194 TI - A large gene cluster for the Clostridium cellulovorans cellulosome. AB - A large gene cluster for the Clostridium cellulovorans cellulosome has been cloned and sequenced upstream and downstream of the cbpA and exgS genes (C.-C. Liu and R. H. Doi, Gene 211:39-47, 1998). Gene walking revealed that the engL gene cluster (Y. Tamaru and R. H. Doi, J. Bacteriol. 182:244-247, 2000) was located downstream of the cbpA-exgS genes. Further DNA sequencing revealed that this cluster contains the genes for the scaffolding protein CbpA, the exoglucanase ExgS, several endoglucanases of family 9, the mannanase ManA, and the hydrophobic protein HbpA containing a surface layer homology domain and a hydrophobic (or cohesin) domain. The sequence of the clustered genes is cbpA-exgS engH-engK-hbpA-engL-man A-engM-engN and is about 22 kb in length. The engN gene did not have a complete catalytic domain, indicating that engN is a truncated gene. This large gene cluster is flanked at the 5' end by a putative noncellulosomal operon consisting of nifV-orf1-sigX-regA and at the 3' end by noncellulosomal genes with homology to transposase (trp) and malate permease (mle). Since gene clusters for the cellulosome are also found in C. cellulolyticum and C. josui, they seem to be typical of mesophilic clostridia, indicating that the large gene clusters may arise from a common ancestor with some evolutionary modifications. PMID- 11004195 TI - Organization and expression of a Thermus thermophilus arginine cluster: presence of unidentified open reading frames and absence of a Shine-Dalgarno sequence. AB - A group of genes regulated by arginine was found clustered in the order argF-ORF1 argC-argJ-ORF4 between other, as yet uncharacterized, open reading frames (ORFs). Transcription starts were identified immediately upstream from argF and ORF4. Arginine repressed transcription that was initiated at argF but induced transcription of ORF4. The functions of ORF1 and ORF4 are unknown, but analysis of the sequence of ORF4 suggests that it is a membrane protein, possibly involved in transport of arginine or a related metabolite. Mobility shift and DNase I footprinting have revealed specific binding of pure Escherichia coli ArgR to the promoter region of Thermus thermophilus argF. These results suggest that argF transcription is controlled by a repressor homologous to those characterized in enteric bacteria and bacilli. Thermus argF mRNA is devoid of Shine-Dalgarno (SD) sequences. However, downstream from the ATG start codon of argF and many other Thermus genes (with or without an SD box), sequences were found to be complementary to nucleotides 1392 to 1409 of Thermus 16S rRNA, suggesting that an mRNA-rRNA base pairing in this region is important for correct translation initiation. PMID- 11004196 TI - Mobilization of poly(3-hydroxybutyrate) in Ralstonia eutropha. AB - Ralstonia eutropha H16 degraded (mobilized) previously accumulated poly(3 hydroxybutyrate) (PHB) in the absence of an exogenous carbon source and used the degradation products for growth and survival. Isolated native PHB granules of mobilized R. eutropha cells released 3-hydroxybutyrate (3HB) at a threefold higher rate than did control granules of nonmobilized bacteria. No 3HB was released by native PHB granules of recombinant Escherichia coli expressing the PHB biosynthetic genes. Native PHB granules isolated from chromosomal knockout mutants of an intracellular PHB (i-PHB) depolymerase gene of R. eutropha H16 and HF210 showed a reduced but not completely eliminated activity of 3HB release and indicated the presence of i-PHB depolymerase isoenzymes. PMID- 11004197 TI - The Streptococcus pneumoniae beta-galactosidase is a surface protein. AB - The beta-galactosidase gene of Streptococcus pneumoniae, bgaA, encodes a putative 2,235-amino-acid protein with the two amino acid motifs characteristic of the glycosyl hydrolase family of proteins. In addition, an N-terminal signal sequence and a C-terminal LPXTG motif typical of surface-associated proteins of gram positive bacteria are present. Trypsin treatment of cells resulted in solubilization of the enzyme, documenting that it is associated with the cell envelope. In order to obtain defined mutants suitable for lacZ reporter experiments, the bgaA gene was disrupted, resulting in a complete absence of endogenous beta-galactosidase activity. The results are consistent with beta galactosidase being a surface protein that seems not to be involved in lactose metabolism but that may play a role during pathogenesis. PMID- 11004198 TI - No genetic barriers between Salmonella enterica serovar typhimurium and Escherichia coli in SOS-induced mismatch repair-deficient cells. AB - Conjugational crosses trigger SOS induction in Escherichia coli F(-) cells mated with Salmonella enterica serovar Typhimurium Hfr donors. Using an epigenetic indicator of SOS induction, we showed that a strong SOS response occurring in a subpopulation of mated mismatch repair-deficient cells totally abolishes genetic barriers between these two genera. PMID- 11004199 TI - On the architecture of the gram-negative bacterial murein sacculus. AB - The peptidoglycan network of the murein sacculus must be porous so that nutrients, waste products, and secreted proteins can pass through. Using Escherichia coli and Pseudomonas aeruginosa as a baseline for gram-negative sacculi, the hole size distribution in the peptidoglycan network has been modeled by computer simulation to deduce the network's properties. By requiring that the distribution of glycan chain lengths predicted by the model be in accord with the distribution observed, we conclude that the holes are slits running essentially perpendicular to the local axis of the glycan chains (i. e., the slits run along the long axis of the cell). This result is in accord with previous permeability measurements of Beveridge and Jack and Demchik and Koch. We outline possible advantages that might accrue to the bacterium via this architecture and suggest ways in which such defect structures might be detected. Certainly, large molecules do penetrate the peptidoglycan layer of gram-negative bacteria, and the small slits that we suggest might be made larger by the bacterium. PMID- 11004200 TI - Identification in Listeria monocytogenes of MecA, a homologue of the Bacillus subtilis competence regulatory protein. AB - We identified in Listeria monocytogenes a gene encoding a protein homologous to MecA, a regulatory protein acting with ClpC and ComK in the competence pathway of Bacillus subtilis. In L. monocytogenes, MecA is involved, along with ClpC and ClpP, in the downregulation of a 64-kDa secreted protein. In B. subtilis, the MecA protein of L. monocytogenes behaves as a regulatory protein, controlling the transcription of comK and comG. Complete or disrupted ComK homologues were also found in L. monocytogenes. However, we failed to detect competence in various strains of L. monocytogenes, including those with intact ComK. Our results suggest that the functions of MecA in the saprophytes L. monocytogenes and B. subtilis have presumably diverged in response to their respective ecological niches. PMID- 11004201 TI - Regulation of exocytosis by protein kinases and Ca(2+) in pancreatic duct epithelial cells. AB - We asked if the mechanisms of exocytosis and its regulation in epithelial cells share features with those in excitable cells. Cultured dog pancreatic duct epithelial cells were loaded with an oxidizable neurotransmitter, dopamine or serotonin, and the subsequent release of these exogenous molecules during exocytosis was detected by carbon-fiber amperometry. Loaded cells displayed spontaneous exocytosis that may represent constitutive membrane transport. The quantal amperometric events induced by fusion of single vesicles had a rapid onset and decay, resembling those in adrenal chromaffin cells and serotonin secreting leech neurons. Quantal events were frequently preceded by a "foot," assumed to be leak of transmitters through a transient fusion pore, suggesting that those cell types share a common fusion mechanism. As in neurons and endocrine cells, exocytosis in the epithelial cells could be evoked by elevating cytoplasmic Ca(2+) using ionomycin. Unlike in neurons, hyperosmotic solutions decreased exocytosis in the epithelial cells, and giant amperometric events composed of many concurrent quantal events were observed occasionally. Agents known to increase intracellular cAMP in the cells, such as forskolin, epinephrine, vasoactive intestinal peptide, or 8-Br-cAMP, increased the rate of exocytosis. The forskolin effect was inhibited by the Rp-isomer of cAMPS, a specific antagonist of protein kinase A, whereas the Sp-isomer, a specific agonist of PKA, evoked exocytosis. Thus, PKA is a downstream effector of cAMP. Finally, activation of protein kinase C by phorbol-12-myristate-13-acetate also increased exocytosis. The PMA effect was not mimicked by the inactive analogue, 4alpha-phorbol-12,13-didecanoate, and it was blocked by the PKC antagonist, bisindolylmaleimide I. Elevation of intracellular Ca(2+) was not needed for the actions of forskolin or PMA. In summary, exocytosis in epithelial cells can be stimulated directly by Ca(2+), PKA, or PKC, and is mediated by physical mechanisms similar to those in neurons and endocrine cells. PMID- 11004202 TI - Time course and Ca(2+) dependence of sensitivity modulation in cyclic GMP-gated currents of intact cone photoreceptors. AB - We determined the Ca(2+) dependence and time course of the modulation of ligand sensitivity in cGMP-gated currents of intact cone photoreceptors. In electro permeabilized single cones isolated from striped bass, we measured outer segment current amplitude as a function of cGMP or 8Br-cGMP concentrations in the presence of various Ca(2+) levels. The dependence of current amplitude on nucleotide concentration is well described by the Hill function with values of K(1/2), the ligand concentration that half-saturates current, that, in turn, depend on Ca(2+). K(1/2) increases as Ca(2+) rises, and this dependence is well described by a modified Michaelis-Menten function, indicating that modulation arises from the interaction of Ca(2+) with a single site without apparent cooperativity. (Ca)K(m), the Michaelis-Menten constant for Ca(2+) concentration is 857 +/- 68 nM for cGMP and 863 +/- 51 for 8Br-cGMP. In single cones under whole-cell voltage clamp, we simultaneously measured changes in membrane current and outer segment free Ca(2+) caused by sudden Ca(2+) sequestration attained by uncaging diazo-2. In the presence of constant 8Br-cGMP, 15 micro, Ca(2+) concentration decrease was complete within 50 ms and membrane conductance was enhanced 2.33 +/- 0.95-fold with a mean time to peak of 1.25 +/- 0.23 s. We developed a model that assumes channel modulation is a pseudo-first-order process kinetically limited by free Ca(2+). Based on the experimentally measured changes in Ca(2+) concentration, model simulations match experimental data well by assigning the pseudo-first-order time constant a mean value of 0.40 +/- 0.14 s. Thus, Ca(2+)-dependent ligand modulation occurs over the concentration range of the normal, dark-adapted cone. Its time course suggests that its functional effects are important in the recovery of the cone photoresponse to a flash of light and during the response to steps of light, when cones adapt. PMID- 11004203 TI - Cysteine modification of a putative pore residue in ClC-0: implication for the pore stoichiometry of ClC chloride channels. AB - The ClC channel family consists of chloride channels important for various physiological functions. Two members in this family, ClC-0 and ClC-1, share approximately 50-60% amino acid identity and show similar gating behaviors. Although they both contain two subunits, the number of pores present in the homodimeric channel is controversial. The double-barrel model proposed for ClC-0 was recently challenged by a one-pore model partly based on experiments with ClC 1 exploiting cysteine mutagenesis followed by modification with methanethiosulfonate (MTS) reagents. To investigate the pore stoichiometry of ClC 0 more rigorously, we applied a similar strategy of MTS modification in an inactivation-suppressed mutant (C212S) of ClC-0. Mutation of lysine 165 to cysteine (K165C) rendered the channel nonfunctional, but modification of the introduced cysteine by 2-aminoethyl MTS (MTSEA) recovered functional channels with altered properties of gating-permeation coupling. The fast gate of the MTSEA modified K165C homodimer responded to external Cl(-) less effectively, so the P(o)-V curve was shifted to a more depolarized potential by approximately 45 mV. The K165C-K165 heterodimer showed double-barrel-like channel activity after MTSEA modification, with the fast-gating behaviors mimicking a combination of those of the mutant and the wild-type pore, as expected for the two-pore model. Without MTSEA modification, the heterodimer showed only one pore, and was easier to inactivate than the two-pore channel. These results showed that K165 is important for both the fast and slow gating of ClC-0. Therefore, the effects of MTS reagents on channel gating need to be carefully considered when interpreting the apparent modification rate. PMID- 11004204 TI - Calcium wave propagation in pancreatic acinar cells: functional interaction of inositol 1,4,5-trisphosphate receptors, ryanodine receptors, and mitochondria. AB - In pancreatic acinar cells, inositol 1,4,5-trisphosphate (InsP(3))-dependent cytosolic calcium ([Ca(2+)](i)) increases resulting from agonist stimulation are initiated in an apical "trigger zone," where the vast majority of InsP(3) receptors (InsP(3)R) are localized. At threshold stimulation, [Ca(2+)](i) signals are confined to this region, whereas at concentrations of agonists that optimally evoke secretion, a global Ca(2+) wave results. Simple diffusion of Ca(2+) from the trigger zone is unlikely to account for a global [Ca(2+)](i) elevation. Furthermore, mitochondrial import has been reported to limit Ca(2+) diffusion from the trigger zone. As such, there is no consensus as to how local [Ca(2+)](i) signals become global responses. This study therefore investigated the mechanism responsible for these events. Agonist-evoked [Ca(2+)](i) oscillations were converted to sustained [Ca(2+)](i) increases after inhibition of mitochondrial Ca(2+) import. These [Ca(2+)](i) increases were dependent on Ca(2+) release from the endoplasmic reticulum and were blocked by 100 microM ryanodine. Similarly, "uncaging" of physiological [Ca(2+)](i) levels in whole-cell patch-clamped cells resulted in rapid activation of a Ca(2+)-activated current, the recovery of which was prolonged by inhibition of mitochondrial import. This effect was also abolished by ryanodine receptor (RyR) blockade. Photolysis of d-myo InsP(3) P(4(5))-1-(2-nitrophenyl)-ethyl ester (caged InsP(3)) produced either apically localized or global [Ca(2+)](i) increases in a dose-dependent manner, as visualized by digital imaging. Mitochondrial inhibition permitted apically localized increases to propagate throughout the cell as a wave, but this propagation was inhibited by ryanodine and was not seen for minimal control responses resembling [Ca(2+)](i) puffs. Global [Ca(2+)](i) rises initiated by InsP(3) were also reduced by ryanodine, limiting the increase to a region slightly larger than the trigger zone. These data suggest that, while Ca(2+) release is initially triggered through InsP(3)R, release by RyRs is the dominant mechanism for propagating global waves. In addition, mitochondrial Ca(2+) import controls the spread of Ca(2+) throughout acinar cells by modulating RyR activation. PMID- 11004205 TI - Pore block versus intrinsic gating in the mechanism of inward rectification in strongly rectifying IRK1 channels. AB - The IRK1 channel is inhibited by intracellular cations such as Mg(2+) and polyamines in a voltage-dependent manner, which renders its I-V curve strongly inwardly rectifying. However, even in excised patches exhaustively perfused with a commonly used artificial intracellular solution nominally free of Mg(2+) and polyamines, the macroscopic I-V curve of the channels displays modest rectification. This observation forms the basis of a hypothesis, alternative to the pore-blocking hypothesis, that inward rectification reflects the enhancement of intrinsic channel gating by intracellular cations. We find, however, that residual rectification is caused primarily by the commonly used pH buffer HEPES and/or some accompanying impurity. Therefore, inward rectification in the strong rectifier IRK1, as in the weak rectifier ROMK1, can be accounted for by voltage dependent block of its ion conduction pore by intracellular cations. PMID- 11004207 TI - Protein translocation across planar bilayers by the colicin Ia channel-forming domain: where will it end? AB - Colicin Ia, a 626-residue bactericidal protein, consists of three domains, with the carboxy-terminal domain (C domain) responsible for channel formation. Whole colicin Ia or C domain added to a planar lipid bilayer membrane forms voltage gated channels. We have shown previously that the channel formed by whole colicin Ia has four membrane-spanning segments and an approximately 68-residue segment translocated across the membrane. Various experimental interventions could cause a longer or shorter segment within the C domain to be translocated, making us wonder why translocation normally stops where it does, near the amino-terminal end of the C domain (approximately residue 450). We hypothesized that regions upstream from the C domain prevent its amino-terminal end from moving into and across the membrane. To test this idea, we prepared C domain with a ligand attached near its amino terminus, added it to one side of a planar bilayer to form channels, and then probed from the opposite side with a water-soluble protein that can specifically bind the ligand. The binding of the probe had a dramatic effect on channel gating, demonstrating that the ligand (and hence the amino-terminal end of the C domain) had moved across the membrane. Experiments with larger colicin Ia fragments showed that a region of more than 165 residues, upstream from the C domain, can also move across the membrane. All of the colicin Ia carboxy-terminal fragments that we examined form channels that pass from a state of relatively normal conductance to a low-conductance state; we interpret this passage as a transition from a channel with four membrane-spanning segments to one with only three. PMID- 11004206 TI - Ion interactions in the high-affinity binding locus of a voltage-gated Ca(2+) channel. AB - The selectivity filter of voltage-gated Ca(2+) channels is in part composed of four Glu residues, termed the EEEE locus. Ion selectivity in Ca(2+) channels is based on interactions between permeant ions and the EEEE locus: in a mixture of ions, all of which can pass through the pore when present alone, those ions that bind weakly are impermeant, those that bind more strongly are permeant, and those that bind more strongly yet act as pore blockers as a consequence of their low rate of unbinding from the EEEE locus. Thus, competition among ion species is a determining feature of selectivity filter function in Ca(2+) channels. Previous work has shown that Asp and Ala substitutions in the EEEE locus reduce ion selectivity by weakening ion binding affinity. Here we describe for wild-type and EEEE locus mutants an analysis at the single channel level of competition between Cd(2+), which binds very tightly within the EEEE locus, and Ba(2+) or Li(+), which bind less tightly and hence exhibit high flux rates: Cd(2+) binds to the EEEE locus approximately 10(4)x more tightly than does Ba(2+), and approximately 10(8)x more tightly than does Li(+). For wild-type channels, Cd(2+) entry into the EEEE locus was 400x faster when Li(+) rather than Ba(2+) was the current carrier, reflecting the large difference between Ba(2+) and Li(+) in affinity for the EEEE locus. For the substitution mutants, analysis of Cd(2+) block kinetics shows that their weakened ion binding affinity can result from either a reduction in blocker on rate or an enhancement of blocker off rate. Which of these rate effects underlay weakened binding was not specified by the nature of the mutation (Asp vs. Ala), but was instead determined by the valence and affinity of the current-carrying ion (Ba(2+) vs. Li(+)). The dependence of Cd(2+) block kinetics upon properties of the current-carrying ion can be understood by considering the number of EEEE locus oxygen atoms available to interact with the different ion pairs. PMID- 11004208 TI - Effect of luminal atrial natriuretic peptide on chloride reabsorption in mouse cortical thick ascending limb: inhibition by endothelin. AB - Insofar as neutral endopeptidase inhibition has afforded evidence for a tubular luminal action of atrial natriuretic peptide (ANP), the present study was undertaken to investigate a possible effect of the peptide on chloride reabsorption (JCl) in thick ascending limb (TAL). Luminal addition of ANP to in vitro microperfused cortical TAL (CTAL) significantly decreased JCl with a threshold and a maximum concentration of 10(-12) M and 10(-9) M, respectively. A similar effect of 10(-9) M ANP was observed in medullary TAL (MTAL). The effect of luminal ANP was significantly reduced by HS-142-1, a specific inhibitor of guanylyl cyclase receptor, and by H-8, a protein kinase G inhibitor, but was not affected by the protein kinase C inhibitor bisindolylmaleimide I. Unexpectedly, the effect of ANP was not additive with that of endothelin (ET), a peptide that was previously shown to decrease JCl in TAL through a calcium-independent, protein kinase C-mediated pathway. Indeed, ET-1 (10(-8) M in the lumen) significantly decreased JCl and prevented a further effect of ANP on the same tubule. Similarly, the decrease of JCl induced by simultaneous addition of ET and ANP was not higher than that obtained with each agent alone. Conversely, the inhibitory effect of ANP was enhanced in the presence of cyclic guanosine monophosphate (cGMP; 10(-6) M in the lumen). ET-1 significantly attenuated the ANP-stimulated generation of cGMP in microdissected CTAL and failed to prevent a further decrease of JCl promoted by a permeant cGMP analogue. It is concluded that luminal ANP decreased Cl reabsorption in mouse CTAL and MTAL. This effect was abrogated by ET-1 as a result of the inhibition of ANP-stimulated cGMP generation. PMID- 11004209 TI - The luminal P2Y receptor in the isolated perfused mouse cortical collecting duct. AB - Extracellular nucleotides regulate renal ion transport. With the use of in vitro perfusion and [Ca(2+)](i) imaging, this study investigated whether mouse and rabbit cortical collecting ducts (CCD) respond to luminal nucleotides. In mouse CCD, luminal ATP (EC(50): 10 microM) and UTP (EC(50): 9.7 microM) increased [Ca(2+)](i) with an initial peak and a plateau. To make certain that basolateral P2 receptors were not activated by luminal nucleotides via leak diffusion, luminal trypsin (1 microM), a known agonist for basolateral proteinase-activated receptors, was perfused. Mouse CCD that were responsive to luminal ATP were nonresponsive to luminal trypsin but always showed [Ca(2+)](i) elevations by basolateral trypsin (10 or 100 nM). Luminal alpha,beta- and beta,gamma-methylene ATP, 2-methyl-S-ATP, ADP, UDP, and 2',3'-O-4-benzoylbenzoyl ATP had no effect (100 microM, n = 9). Without external Ca(2+), luminal ATP still stimulated a [Ca(2+)](i) increase. Mouse CCD also responded to basolateral ATP (EC(50): 23 microM) and UTP (EC(50): 23 microM) with smaller [Ca(2+)](i) elevations. Confocal microscopy of perfused CCD showed that luminal ATP (100 microM) rapidly increased [Ca(2+)](i) in nearly all cells (n = 6) and the same cells that responded to luminal ATP responded to basolateral ATP (100 microM). In contrast, rabbit CCD did not respond to luminal ATP/UTP (n = 8) despite ATP's known effect from the basolateral side (EC(50): 34 microM). These data indicate the expression of luminal P2Y receptors (probably P2Y(2)) in principal cells of mouse CCD but not in rabbit CCD. PMID- 11004210 TI - Inducible nitric oxide synthase attenuates endothelium-dependent renal microvascular vasodilation. AB - Previous studies have demonstrated that inducible nitric oxide synthase (iNOS) plays a key pathophysiologic role during sepsis. The present study was designed to delineate the consequences of iNOS activation on renal microvascular function. Male Sprague-Dawley rats were given intraperitoneal injections of lipopolysaccharide (LPS; 4 mg/kg) at 16 h and 4 h before experimentation. Afferent and efferent arteriolar diameters from LPS-treated and control rats were assessed in vitro with the use of the blood perfused juxtamedullary nephron technique. Basal afferent and efferent arteriolar diameters of LPS-treated rats averaged 19.7 +/- 0.9 (n = 7) and 18.3 +/- 1.0 microm (n = 5), respectively, and were similar to those of control rats (20.8 +/- 0.3 [n = 6] and 18.4 +/- 0.6 microm [n = 6], respectively). Superfusion with the selective iNOS inhibitor S,S' (1,3-phenylenebis[1,2-ethanediyl]) bisisothiourea (PBIT), at the doses of 0.01, 0.1, and 1 microM, significantly decreased afferent and efferent arteriolar diameters in a dose-dependent manner, whereas afferent or efferent arteriolar diameters of control rats were not altered in response to the same doses of PBIT. In the second series of experiments, superfusion with 10 microM acetylcholine (ACh) significantly increased afferent and efferent arteriolar diameters of LPS treated rats by 14.9 +/- 1.6% (n = 9) and 6.6 +/- 1.1% (n = 6), respectively. The ACh-induced afferent and efferent arteriolar dilator responses were inhibited by superfusion with the nonselective NOS inhibitor N:(omega)-nitro-L-arginine (100 microM). However, afferent and efferent arteriolar dilator responses to ACh were significantly enhanced during selective iNOS inhibition with 1 microM PBIT (40.1 +/- 0.7% and 25.2 +/- 1.3%, respectively). These results suggest that activation of iNOS by LPS increases the influence of nitric oxide on afferent and efferent arteriolar tone and impairs endothelium-dependent nitric oxide effects. PMID- 11004211 TI - Renin inhibition improves pressure natriuresis in essential hypertension. AB - Pressure natriuresis (PN), i.e., a rise in renal sodium excretion in response to a higher BP, is involved in long-term BP regulation. PN is blunted in essential hypertension, but the mechanism is unknown. This study assessed the role of the renin-angiotensin-aldosterone system (RAAS) in PN in eight essential hypertensive men from the individual correlations between spontaneous fluctuations in BP and time corresponding changes in sodium excretion (collected at 2- and 4-h intervals for 48 h), during strict sodium balance, without treatment, and during renin inhibition (remikiren, 600 mg oral compound). Without treatment, daily values for mean arterial pressure were 109.5 +/- 1.9 and 107 +/- 1.9 mmHg, for urinary sodium excretion were 37.2 +/- 2.8 and 42.0 +/- 2.8 mmol/24 h, and for plasma renin activity were 2.34 +/- 0.48 and 2.23 +/- 0.44 nmol/L per h, respectively, for two consecutive days. During remikiren treatment, mean arterial pressure was 101.9 +/- 1.7 and 100.8 +/- 1. 7 mmHg (P: < 0.05, versus baseline). Urinary sodium excretion was 39. 3 +/- 3.7 and 45.2 +/- 5.3 mmol/24 h (not significant versus baseline), and plasma renin activity was 0.79 +/- 0.11 and 0.82 +/- 0.13 nmol/L per h (P: < 0.05 versus baseline). During remikiren treatment, BP correlated positively with sodium excretion in all patients but in only three of eight patients without treatment. The slope of the regression equation was steeper during remikiren treatment in seven of eight patients. Thus, the relationship between BP and natriuresis was more readily apparent during RAAS blockade, suggesting that RAAS activity blunts PN in hypertensive patients. Improved PN may contribute to the hypotensive effect of RAAS blockade and to maintenance of sodium balance at a lower BP level without volume expansion. PMID- 11004212 TI - Stimulation of NADPH oxidase by oxidized low-density lipoprotein induces proliferation of human vascular endothelial cells. AB - Oxidized low-density lipoprotein (OxLDL) exerts proliferation and apoptosis in vascular cells, depending on its concentration and the duration of exposure. Recent studies indicate that [O(2)](-) is involved in cell cycle regulation and that OxLDL stimulates endothelial cells to produce [O(2)](-). This study examined the role of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase as a potential source for [O(2)](-) in the proliferation-inducing activity of OxLDL in cultured human umbilical vein endothelial cells (HUVEC). Human LDL was oxidized by Cu(++), and proliferation of HUVEC was detected by 3H-thymidine incorporation. OxLDL (5 microg/ml) caused an increase in proliferation of HUVEC of 250 to 300%. OxLDL-induced proliferation was blocked by addition of the antioxidants superoxide dismutase and catalase, suggesting that enhanced [O(2)](-) formation was involved. Diphenylene iodonium (DPI, 1 microM), an inhibitor of NADPH oxidase, also prevented OxLDL-induced proliferation of HUVEC, indicating that NADPH oxidase was the source for enhanced [O(2)](-) formation. The OxLDL effect was mimicked by lysophosphatidylcholine (LPC, 10 microM), a compound formed during oxidation of LDL. LPC-induced proliferation was also prevented by coincubation with DPI. Treatment of HUVEC with [O(2)](-) generated by the xanthine/xanthine oxidase reaction resulted in proliferation as did treatment with OxLDL. As expected, this stimulation could not be blocked by DPI. With the use of the cytochrome c-assay, it was demonstrated that OxLDL and LPC enhanced [O(2)](-) formation in HUVEC (by factor 3.2 and by factor 3.5, respectively). Supporting the assumption that NADPH oxidase was the enzyme responsible for [O(2)](-) formation, cells transfected with antisense oligonucleotides for NADPH oxidase showed a significantly reduced [O(2)](-) formation after stimulation with OxLDL and LPC. OxLDL and its compound LPC induce proliferation of HUVEC through activation of NADPH oxidase. The active NADPH oxidase generates [O(2)](-), which mediates the proliferative effects. PMID- 11004213 TI - Expression of LOX-1, an oxidized low-density lipoprotein receptor, in experimental hypertensive glomerulosclerosis. AB - Oxidized low-density lipoprotein (OxLDL) has been implicated in atherosclerosis and glomerulosclerosis. LOX-1 is a recently identified OxLDL receptor that is abundantly expressed in vascular endothelial cells. The aim of the present study was to investigate LOX-1 expression in the kidneys of hypertensive rats. Dahl salt-sensitive (DS) and salt-resistant (DR) rats were fed a 0.3% or 8% NaCl diet. Some DS 8% rats were treated with manidipine or hydralazine. LOX-1 gene expression was markedly elevated in the kidneys and glomeruli of hypertensive DS 8% rats compared with those of normotensive DR and DS 0.3% rats. Prolonged salt loading further increased the renal LOX-1 expression in DS rats. The LOX-1 upregulation in DS 8% rats was accompanied by renal overexpression of transforming growth factor-beta 1 and type I collagen, impaired renal function, and histologic glomerulosclerotic changes, all of which were ameliorated by antihypertensive treatment. LOX-1 was indeed expressed in the glomeruli in vivo and in cultured glomerular cells in vitro. However, LOX-1 expression was elevated in the aorta but not the kidneys of spontaneously hypertensive rats, which exhibited hypertension but minor glomerulosclerotic changes. In conclusion, the LOX-1 upregulation in the kidney of DS 8% rats was parallel to glomerulosclerotic changes and renal dysfunction, suggesting a possible pathogenetic role for renal LOX-1 in the progression to hypertensive glomerulosclerosis. PMID- 11004214 TI - Development of autosomal recessive polycystic kidney disease in BALB/c-cpk/cpk mice. AB - Autosomal recessive polycystic kidney disease (ARPKD) is a rare but devastating inherited disease in humans. Various strains of mice that are homozygous for the cpk gene display renal pathology similar to that seen in human ARPKD. The PKD progresses to renal insufficiency, azotemia, and ultimately a uremic death by approximately 3 wk of age. This study characterizes PKD in mice that are homozygous for the cpk gene on a BALB/c inbred mouse background. The BALB/c cpk/cpk murine model displays renal as well as extrarenal pathology similar to that found in human ARPKD. The renal pathology includes the well-characterized early proximal tubule and, later, massive collecting duct cysts. The extrarenal defects in this murine model include common bile duct dilation, intrahepatic biliary duct cysts with periductal hyperplasia, and pancreatic dysplasia with cysts. Renal mRNA expression of c-myc, a proto-oncogene, and clusterin (SGP-2), a marker associated with immature collecting ducts, decreases during normal development but is upregulated in murine ARPKD. Expression of epidermal growth factor (EGF) mRNA is significantly diminished, whereas EGF receptor mRNA is upregulated in the BALB/c-cpk/cpk kidney compared with phenotypically normal littermates. To determine whether the altered EGF expression contributes to the development of PKD, neonatal mice were treated with exogenous EGF (1 microg/g body wt injected subcutaneously on postnatal days 3 through 9). EGF treatment reduced the relative kidney weight and common bile duct dilation and downregulated renal expression of clusterin and EGF receptor. However, exogenous EGF did not affect the degree of renal failure, the pancreatic pathology, or the misregulated renal expression of c-myc. In summary, the present study characterizes the renal and extrarenal pathology in the BALB/c-cpk/cpk murine model of ARPKD. Renal mRNA expression of EGF is diminished in this mouse model. EGF treatment did not prevent renal failure but ameliorated pathologic changes in the kidney and the biliary ducts of the BALB/c-cpk/cpk mouse. PMID- 11004215 TI - Expression of endothelial nitric oxide synthase in human peritoneal tissue: regulation by Escherichia coli lipopolysaccharide. AB - Changes in the expression of endothelial nitric oxide synthase (eNOS) in the peritoneum could be involved in the peritoneal dysfunction associated with peritoneal inflammation. Demonstrated recently in bovine endothelial cells was the existence of cytosolic proteins that bind to the 3'-untranslated region (3' UTR) of eNOS mRNA and could be implicated in eNOS mRNA stabilization. The present work demonstrates that eNOS protein is expressed in human endothelial and mesothelial peritoneal cells. Escherichia coli lipopolysaccharide shortened the half-life of eNOS message, reducing eNOS protein expression in peritoneal mesothelial and endothelial cells. Moreover, under basal conditions, human peritoneal samples expressed cytosolic proteins that bind to the 3'-UTR of eNOS mRNA. The cytosolic proteins that directly bind to 3'-UTR were identified as a 60 kD protein. After incubation of human peritoneal samples with lipopolysaccharide, the binding activity of the cytosolic 60-kD protein increased in a time-dependent manner. Studies are now necessary to determine the involvement of this 60-kD protein in the regulation of eNOS expression in peritoneal cells and particularly its involvement in the peritoneal dysfunction associated with inflammatory reactions. PMID- 11004216 TI - 1,25-Dihydroxy-19-nor-vitamin D(2), a vitamin D analog with reduced bone resorbing activity in vitro. AB - 1,25-Dihydroxy-19-nor-vitamin D(2) (19-norD(2)), a new analog of 1,25(OH)(2)D(3), suppresses parathyroid hormone in renal failure patients and in uremic rats but has less calcemic activity than 1,25(OH)(2)D(3). Although 19-norD(2) has high affinity for the vitamin D receptor and similar pharmacokinetics to those of 1,25(OH)(2)D(3), it has much less bone resorbing activity in vivo. The intrinsic activity of 19-norD(2) on osteoclastogenesis and activation of bone resorption in mouse bone marrow cultures was examined to determine the mechanism involved. 19 norD(2) and 1,25(OH)(2)D(3) (10 nM) were equivalent in stimulating the formation and maintenance of large multinucleated, tartrate-resistant acid phosphatase positive cells. However, the amount of bone resorbed by osteoclasts stimulated by 10 nM 19-norD(2), as measured by pit-forming assays, was reduced 62% compared with 10 nM 1,25(OH)(2)D(3)-stimulated osteoclasts (P < 0. 05). This difference could not be attributed to enhanced catabolism or to downregulated vitamin D receptor. The rate of degradation of 19-norD(2) in cultures was approximately 20% greater than 1, 25(OH)(2)D(3), not enough to account for the different effects on bone resorption. The VDR levels were identical in cultures that were treated with 19-norD(2) and 1,25(OH)(2)D(3). In summary, 19-norD(2) is less effective than 1,25(OH)(2)D(3) in stimulating mouse marrow osteoclasts to resorb bone. The reason for this difference is not clear but seems to involve the late maturation and/or activation of osteoclasts as the number of pits produced by each tartrate resistant acid phosphatase-positive cell is reduced under stimulation by 19 norD(2) compared with 1,25(OH)(2)D(3). PMID- 11004217 TI - The in vitro effect of calcitriol on parathyroid cell proliferation and apoptosis. AB - Calcitriol treatment is used to reduce parathyroid hormone levels in azotemic patients with secondary hyperparathyroidism (HPT). Whether long-term calcitriol administration reduces parathyroid gland size in patients with severe secondary hyperparathyroidism is not clear. The aim of the study was to evaluate in vitro the effect of calcitriol on parathyroid cell proliferation and apoptosis in normal parathyroid glands and in adenomatous and hyperplastic human parathyroid glands. Freshly harvested parathyroid glands from normal dogs and hyperplastic and adenomatous glands from patients with secondary (2 degrees) and primary (1 degree) HPT undergoing parathyroidectomy were studied. Flow cytometry was used to quantify the cell cycle and apoptosis of parathyroid cells. Apoptosis was also evaluated by DNA electrophoresis and light and electron microscopy. In normal dog parathyroid glands, culture with calcitriol (10(-10) to 10(-7) M) for 24 h produced a dose-dependent inhibitory effect on the progression of cells into the cell cycle and into apoptosis. When glands from patients with 2 degrees HPT were cultured for 24 h, only high calcitriol concentrations (10(-7) M) inhibited the progression through the cell cycle and the induction of apoptosis. In parathyroid adenomas (1 degrees HPT), even a high concentration of calcitriol (10(-7) M) had no significant effect on the cell cycle or apoptosis. The present study shows that in vitro, calcitriol inhibits in a dose-dependent manner in normal parathyroid glands both parathyroid cell proliferation and apoptosis. However, in secondary hyperplasia, only high concentrations of calcitriol inhibited cell proliferation and apoptosis. In 1 degree HPT, even high concentrations of calcitriol had no effect. Because calcitriol simultaneously inhibits both cell proliferation and apoptosis, a reduction in the parathyroid gland mass may not occur as a direct effect of calcitriol treatment. PMID- 11004218 TI - Urinary aquaporin 2 and calciuria correlate with the severity of enuresis in children. AB - This study examined the hypothesis that nocturnal enuresis might be paralleled by aquaporin 2 (AQP2) urinary excretion. Eighty children who experienced nocturnal enuresis were studied and compared with 9 healthy children. The 24-h urine samples were divided into two portions: night collections and day collections. Creatinine equivalents of urine samples from each patient were analyzed by Western blotting. AQP2 levels were semiquantified by densitometric scanning and reported as a ratio between the intensity of the signal in the day urine sample versus the night urine sample (D/N AQP2 ratio). The D/N AQP2 ratio was 0.59 +/- 0.11 (n = 9) in healthy children and increased to 1.27 +/- 0.24 (n = 10) in a subpopulation of enuretic children who had low nocturnal vasopressin levels. In enuretic children who displayed hypercalciuria and had normal vasopressin levels, the D/N AQP2 ratio was 1.05 +/- 0.27 (n = 8). These data indicate that reduced secretion of vasopressin and absorptive hypercalciuria are independently associated with an approximately twofold increase in the urinary D/N AQP2 ratio. When low nocturnal vasopressin levels were associated with hypercalciuria, a nearly threefold increase in the D/N AQP2 ratio was observed (1. 67 +/- 0.41, n = 11). In addition, in all enuretic patients tested, the urinary D/N AQP2 ratio correlates perfectly with the severity of the disorder (nocturnal polyuria). The findings reported in this article indicate that urinary AQP2 correlates with the severity of enuresis in children. PMID- 11004219 TI - Urinary albumin excretion is associated with renal functional abnormalities in a nondiabetic population. AB - Microalbuminuria (MA) is an important early sign of diabetic nephropathy. Hyperfiltration and impaired filtration in relation to albuminuria has been well investigated in diabetic subjects. This study tested the hypothesis that an increased urinary albumin excretion (UAE) is associated with renal functional abnormalities also in nondiabetic subjects. The relation between UAE and creatinine clearances (Ccr) in 7728 nondiabetic subjects was studied. Subjects were divided in four groups according to UAE (mg/24 h): 0 to 15 (control), 15 to 30 (high-normal albuminuria [HNA]), 30 to 300 (MA), >300 (macroalbuminuria). An elevated filtration and a diminished filtration were defined as a Ccr exceeding or below 2x the SD of the control group corrected for age and gender. Ccr followed a parabolic trend, with a higher Ccr in the HNA as compared with control and a lower Ccr in the MA and macroalbuminuria group as compared with HNA. With each increasing UAE level, male sex, age, body mass index, minimal waist circumference, systolic and diastolic BP, plasma glucose, and a positive family history for diabetes all followed a significant linear increasing trend (P < 0.001). After adjustment for age, gender, body mass index, plasma glucose, a positive family history for diabetes, systolic and diastolic BP, antihypertensive medication, and smoking in a multivariate analysis, HNA and MA were independently associated with an elevated filtration (RR 1.8 [95% confidence interval, 1.30 to 2.51] and 1.7 [1.17 to 2. 45]). Macroalbuminuria was independently associated with a diminished filtration (4.3 [range, 1.97 to 9.36]). In conclusion, an elevated UAE might be an important and early sign for progressive renal function loss in a nondiabetic population. PMID- 11004220 TI - Role of lipoprotein (a) and TGF-beta 1 in atherosclerosis of hemodialysis patients. AB - Atherosclerotic vascular disease is a major cause of death for uremic patients who are on hemodialysis (HD). Recent evidence suggests that lipoprotein (a) [Lp(a)] may aggravate atherosclerosis by inhibiting activation of transforming growth factor-beta 1 (TGF-beta 1). Plasma Lp(a) and plasma TGF-beta 1 activation in HD patients (n = 51), chronic renal failure patients not subjected to hemodialysis (non-HD-CRF; n = 12), and healthy volunteers (control; n = 13) were investigated. Plasma Lp(a) was significantly higher in HD (18.75 +/- 1.62 mg/ml) and non-HD-CRF patients (25.0 +/- 8.4 mg/ml) than in control subjects (10.9 +/- 5.8 mg/ml). The degree of atherosclerosis in HD patients was assessed by measuring the intima-media thickness (IMT) and plaque score with the use of an ultrasound scanner. IMT and plaque score were higher in HD and non-HD-CRF patients than in controls. A significant positive correlation was found in HD patients between Lp(a) and IMT (r = 0. 377, P < 0.01) as well as between Lp(a) and plaque score (r = 0.43, P < 0.01). Plasma total TGF-beta 1 significantly increased in HD (119.8 +/- 53.5 ng/ml) and non-HD-CRF patients (93.2 +/- 25.0 ng/ml) compared with control subjects (17.7 +/- 6.4 ng/ml), whereas the plasma level of mature (active) TGF-beta1 did not differ among the groups. When plasma TGF-beta 1 and supernatant TGF-beta 1 from cultured peripheral mononuclear cells were compared before and after an HD session, neither total nor mature TGF-beta 1 showed a significant difference between the values before and after an HD session. There were no significant relationships between plasma total TGF-beta 1 and IMT or plaque score, between mature TGF-beta 1 and IMT or plaque score, or between mature TGF-beta 1 and Lp(a). In conclusion, Lp(a) may be an important atherogenic factor in CRF patients. However, it was not clarified whether Lp(a) exerts its effect by inhibiting TGF-beta 1 activation in CRF patients. PMID- 11004221 TI - Prognosis of anti-hepatitis C virus antibody-positive patients on regular hemodialysis therapy. AB - The prevalence of hepatitis C virus (HCV) infection is high in patients who are on chronic hemodialysis, but the role of HCV infection and HCV-related liver disease in the mortality of these patients has not been shown. Therefore, we conducted a prospective cohort study of 1470 patients who were on chronic hemodialysis (17 to 89 yr old) from 16 dialysis centers in Japan. Among them, 276 patients (18.8%) were positive for anti-HCV antibodies and 1194 patients were negative. The patients were followed for 6 yr from 1993 to 1999. Only one case, a patient from the anti-HCV-antibody-positive group, was lost to the follow-up during this period. The mortality was higher in the anti-HCV-antibody-positive group (91 of 276 patients died) than in the anti-HCV-antibody-negative group (277 of 1193 died) (33.0% versus 23.2%, P< 0.01). A Cox proportional hazard examination showed that positivity for anti-HCV antibodies was one of the risk factors for death with an adjusted relative risk of 1.57 (95% confidence interval, 1.23 to 2.00). As a cause of death, hepatocellular carcinoma and liver cirrhosis were significantly more frequent in the anti-HCV-antibody-positive patients than in the anti-HCV-antibody-negative patients (5.5% versus 0.0%, P< 0.001; 8.8% versus 0.4%, P< 0.001, respectively). These findings show that the mortality is increased in anti-HCV-antibody-positive patients who are on chronic hemodialysis. Hepatocellular carcinoma and liver cirrhosis are factors that may influence the mortality. PMID- 11004222 TI - Avoidance of cyclosporine in renal transplantation: effects of daclizumab, mycophenolate mofetil, and steroids. AB - Cyclosporine (CsA) has been implicated in both acute and chronic graft dysfunction. The addition of humanized IL-2 receptor antibody daclizumab (DZB) to CsA-based immunosuppression decreases the rate of acute renal transplant rejection. Therefore, 45 patients were evaluated in an immunosuppressive protocol that included DZB, mycophenolate mofetil (MMF), and steroids without CsA. This was a prospective, nonrandomized, open-label trial of the efficacy and safety of the treatment. DZB was given intravenously at 2 mg/kg before transplantation and then at 1 mg/kg every 2 wk for four doses, MMF was given orally at 3 g/d, and methylprednisolone/prednisone was given at 7 mg/kg per day and tapered to 15 mg/d at 6 mo. CsA was added to the regimen when patients developed acute rejection episodes or adverse effects to steroids or MMF; 49% of patients were spared CsA maintenance. Patients without CsA had lower serum creatinine at 6 mo and needed fewer medications to control BP. Incidence of biopsyproven rejections was 31% and occurred early (median, 10 d). These rejection episodes occurred earlier in cadaver transplants (median, 7 d) and later in living donor transplants (median, 62 days). Acute rejections occurred at a higher frequency (46% versus 34%) and earlier (6.5 versus 15 d) in patients with delayed graft function compared with patients without delayed graft function. Most of the rejections were moderate and easily reversible. The actuarial 1-yr graft survival was 95% with 100% patient survival. PMID- 11004223 TI - A meta-analysis of immunosuppression withdrawal trials in renal transplantation. AB - Since the publication of previous meta-analyses of cyclosporine (CsA) and prednisone withdrawal in renal transplant recipients, several additional randomized controlled trials with longer follow-up have been reported. Currently, in nine prednisone withdrawal trials (n = 1461), the proportion of patients with acute rejection was increased by 0.14 (95% confidence interval = 0.10 to 0. 17, P < 0.001). In nine prednisone withdrawal trials (n = 1899), the relative risk (RR; RR = 1.0 indicates no risk) of graft failure after withdrawal was also increased (RR = 1.40; range, 1.09 to 1.70, P = 0.012). There was no evidence of between study heterogeneity for either acute rejection or graft failure in the prednisone withdrawal trials by a chi(2) test (P > 0.05). In 10 CsA withdrawal trials (n = 1049), the proportion of patients with acute rejection was increased by 0.11 (0.07 to 0.15, P < 0.001). In 12 trials (n = 1151), the RR of graft failure after CsA withdrawal was 1.06 (95% confidence interval, 0.82 to 1.29, P = 0.646), but a chi(2) test indicated that there was study heterogeneity. However, there was no evidence of heterogeneity in the six studies (n = 632) with at least 4.0 yr (5.8 +/- 1.7) of follow-up (RR = 0.92; range, 0.64 to 1.20, P = 0.569) or in the seven trials (n = 962) published in peer-reviewed journals (RR = 0.95; range, 0.70 to 1.20 P = 0.682). Finally, in three trials (n = 259) that compared CsA and prednisone withdrawal, there was a nonsignificant trend for less graft failure with CsA withdrawal (RR = 0.63; range, 0.08 to 1.16, P = 0.190). Thus, unlike prednisone withdrawal, CsA withdrawal in select patients seems to impart little risk of long-term graft failure. PMID- 11004224 TI - Effect of MTHFR 1298A-->C and MTHFR 677C-->T genotypes on total homocysteine, folate, and vitamin B(12) plasma concentrations in kdiney graft recipients. AB - The effect of 5,10-methylenetetrahydrofolate reductase (MTHFR) 677C-->T and 1298A ->C on total homocysteine (tHcy), folate and vitamin B(12) levels was investigated in 733 kidney graft recipients. The six major genotype combinations were used as grouping variables, and age, gender, BMI, serum creatinine, and creatinine clearance and ln-folate, ln-vitamin B(12), or logarithmus naturalis tHcy (ln-tHcy) were used as covariates in three ANCOVA and multiple stepwise linear regression models. Hyperhomocysteinemia was present in 49.7% of the patients. The allele frequency of MTHFR 677T and 1298C was 0.319 and 0.326. MTHFR genotype and all other variables were significant predictors of ln-tHcy (higher tHcy plasma levels for MTHFR 677TT/1298AA versus all other five genotype groups: P < 0. 05). BMI, creatinine clearance, ln-tHcy, and MTHFR genotype influenced ln folate (lower folate levels for MTHFR 677TT/1298AA versus all other genotype groups: P < 0.05). Creatinine clearance and ln-tHcy were the only predictors of ln-vitamin B(12) levels. In a prespecified subgroup analysis (n = 496), the MTHFR genotype also influenced tHcy levels and compound heterozygous patients had significantly lower folate levels as compared with MTHFR 677CC/1298AA and 677CC/1298CC. This study shows that the MTHFR 677TT/1298AA and 677CT/1298AC genotypes are significant predictors of tHcy and folate plasma levels. PMID- 11004225 TI - Altered expression of type II sodium/phosphate cotransporter in polycystic kidney disease. AB - Renal phosphate (Pi) absorption is mediated via the type II sodium/Pi cotransporter (NaPi-2) in the brush border membrane (BBM) of proximal tubules. Simultaneous detection of NaPi-2 mRNA by in situ hybridization and of NaPi-2 immunoreactivity by immunohistochemistry was performed to investigate the distribution of the cotransporter in healthy control rats and during progression of autosomal dominant polycystic kidney disease (ADPKD). The purpose of the study was to disclose a relation between proximal tubular cell differentiation and NaPi 2 expression. In controls, NaPi-2 expression was present in the entire proximal tubule. In the Han:SPRD (cy/+) model for ADPKD, the proximal nephron is primarily affected by the cystic changes. Epithelial proliferation and impaired epithelial matrix interaction result in a loss of cell differentiation that eventually leads to cystic enlargement of the nephron. Normal expression of NaPi-2 in this model was found only in tubules with intact BBM. Loss of BBM and cellular interdigitation were paralleled by the loss of NaPi-2 in situ hybridization and immunoreactive signals. These changes were moderate and focal in 2-mo-old rats and generalized all over the cortex after 8 mo. Advanced renal damage in the older PKD group was associated with mild phosphaturia, which suggests functional insufficiency of tubular NaPi-2 reabsorption. These data show how proliferative changes and loss of tubular epithelial differentiation in ADPKD may prevent functional expression of the NaPi-2 system in the proximal tubule in a rapidly progressive manner. NaPi-2 in proximal tubule BBM is suggested to play an important role in impaired tubular absorption of Pi in renal disease. PMID- 11004226 TI - Cyclosporine avoidance. PMID- 11004227 TI - Inherited disorders of renal magnesium handling. AB - The genetic basis and cellular defects of a number of primary magnesium wasting diseases have been elucidated over the past decade. This review correlates the clinical pathophysiology with the primary defect and secondary changes in cellular electrolyte transport. The described disorders include (1) hypomagnesemia with secondary hypocalcemia, an earlyonset, autosomal-recessive disease segregating with chromosome 9q12-22.2; (2) autosomal-dominant hypomagnesemia caused by isolated renal magnesium wasting, mapped to chromosome 11q23; (3) hypomagnesemia with hypercalciuria and nephrocalcinosis, a recessive condition caused by a mutation of the claudin 16 gene (3q27) coding for a tight junctional protein that regulates paracellular Mg(2+) transport in the loop of Henle; (4) autosomal-dominant hypoparathyroidism, a variably hypomagnesemic disorder caused by inactivating mutations of the extracellular Ca(2+)/Mg(2+) sensing receptor, CASR: gene, at 3q13.3-21 (a significant association between common polymorphisms of the CASR: and extracellular Mg(2+) concentration has been demonstrated in a healthy adult population); and (5) Gitelman syndrome, a recessive form of hypomagnesemia caused by mutations in the distal tubular NaCl cotransporter gene, SLC12A3, at 16q13. The basis for renal magnesium wasting in this disease is not known. These inherited conditions affect different nephron segments and different cell types and lead to variable but increasingly distinguishable phenotypic presentations. No doubt, there are in the general population other disorders that have not yet been identified or characterized. The continued use of molecular techniques to probe the constitutive and congenital disturbances of magnesium metabolism will increase the understanding of cellular magnesium transport and provide new insights into the way these diseases are diagnosed and managed. PMID- 11004228 TI - Coronary heart disease in chronic renal insufficiency: some management considerations. PMID- 11004229 TI - New hope for the treatment of pulmonary hypertension: novel approaches to a complex disease. PMID- 11004230 TI - Fatty acid synthase inhibitors reduce food intake and body weight. PMID- 11004231 TI - Changes in expression of platelet-derived growth factor and its receptors in the lungs of newborn rats exposed to air or 60% O(2). AB - PDGF-related gene expression has been well characterized during fetal rat lung development and adult rat lung injury, but not during normal postnatal lung growth or injury. Lung expression of the mRNA for PDGF-A, -B, -alpha R, and -beta R and immunoreactive PDGF-AA, -BB, -alpha R, and -beta R were assessed in rat pups raised in air or 60% O(2) for up to 14 d after birth. Expression of mRNA and immunoreactive ligand did not correlate for pups raised in air. Immunoreactive PDGF-alpha R and -beta R, but not PDGF-AA and -BB, were evident throughout the lung at birth. Both PDGF-AA and -BB were evident in airway epithelium, PDGF-BB in alveolar epithelial cells and PDGF-AA was widely distributed in parenchymal tissue at 4 d. PDGF-alpha R was localized to airway epithelium, and PDGF-beta R to subendothelial perivascular regions and to airway and alveolar epithelium at 4 d. Immunoreactive PDGF ligands all declined after 4 d. Intraperitoneal injection of neutralizing antibodies or truncated soluble receptors to PDGF-BB reduced lung DNA synthesis in air. Exposure to 60% O(2) significantly increased mRNA for PDGF B, -beta R, and -alpha R, but not PDGF-A, relative to air-exposed lung at various time points after birth. PDGF-A, -B, and -alpha R immunoreactivities in these lungs were reduced and delayed, consistent with a global inhibition of lung growth. Pups exposed to 60% O(2) had a similar distribution of PDGF-beta R to that seen in air, except that at 14 d PDGF-beta R was distributed throughout the lung parenchyma. We conclude that PDGF ligands and receptors are important for normal postnatal lung growth and that their expression is delayed by O(2) exposure. PMID- 11004232 TI - Effects of retinoic acid on airspace development and lung collagen in hyperoxia exposed newborn rats. AB - Impaired septal formation and decreased alveolarization are often caused by hyperoxic injury to the developing lung and are characteristic features of bronchopulmonary dysplasia. Dexamethasone, frequently administered to infants during oxygen exposure, also inhibits septal formation in the newborn lung. Vitamin A administration reduces the incidence of bronchopulmonary dysplasia in vitamin A-deficient premature infants, and retinoic acid improves alveolarization in newborn rats treated with dexamethasone, indicating that retinoic acid may be useful in preventing hyperoxia-induced impaired septation in bronchopulmonary dysplasia. To investigate whether treatment with retinoic acid during exposure to hyperoxia would improve septal formation, newborn rats exposed to > or =90% O(2) from d 3 of life to d 14 were treated with retinoic acid (d 3-13 of life) and/or dexamethasone (d 4-13 of life). In contrast with the effects of retinoic acid on dexamethasone-induced inhibition of alveolarization, we found that retinoic acid did not improve septal formation or decrease airspace size in animals exposed to hyperoxia alone or to hyperoxia plus dexamethasone. Retinoic acid did, however, increase collagen in airspace walls as demonstrated by staining and immunohistochemistry. There was no increase in procollagen mRNA by Northern hybridization analysis, indicating that retinoic acid-associated increases in lung collagen are likely due to posttranscriptional regulation. There was a trend toward increased survival in hyperoxia in animals treated with retinoic acid to the extent that combined therapy with retinoic acid and dexamethasone resulted in the greatest improvement in animal survival. These results suggest that although retinoic acid may be of benefit in hyperoxia-induced lung injury and may have important effects on lung matrix, it does not prevent impairment of septation or induce alveolar formation during exposure to hyperoxia. PMID- 11004233 TI - Noninvasive detection of changes in cerebral blood flow by near-infrared spectroscopy in a piglet model of hydrocephalus. AB - Formulation of rational interventions in infantile hydrocephalus is limited by the inability to monitor cerebral hemodynamics quantitatively, continuously, and noninvasively. Near-infrared spectroscopy (NIRS) measures changes in cerebral concentration of oxygenated and deoxygenated hemoglobin (HbO(2) and Hb); HbD is the derived difference between HbO(2) and Hb. Our previous work showed that HbD reflected cerebral blood flow (CBF) measured by radioactive microspheres in a piglet model of systemic hypotension. This study was designed to determine whether NIRS detected important changes in cerebral perfusion and oxygenation in a piglet model of hydrocephalus and whether changes in HbD accurately reflected changes in CBF. Acute hydrocephalus was produced in neonatal piglets by intraventricular infusion of "mock cerebrospinal fluid." Intracranial pressure (ICP) was maintained for several minutes at approximately 10, 20, and 30 mm Hg above the baseline ICP. CBF was measured in cerebral cortex, white matter, and basal ganglia at each ICP by radioactive microspheres. Changes in HbO(2) and Hb were measured continuously by NIRS. Cerebral perfusion pressure declined with increasing ICP, and this decline was accompanied by significant decreases in HbD measured by NIRS and CBF measured by radioactive microspheres. There was a strong correlation between changes in HbD and individual changes in CBF in cerebral cortex, white matter, and basal ganglia (all p < 0.0001). This study demonstrates that changes in HbD reflect changes in CBF over a wide range of ICP in a model of acute hydrocephalus. This reproducible and easily obtained measurement by NIRS could facilitate considerably decisions concerning therapeutic interventions. PMID- 11004234 TI - Evolution in the hypervariable region of hepatitis C virus in infants after vertical transmission. AB - To elucidate the clonal evolution of hepatitis C virus (HCV) during mother-to infant transmission, we prospectively analyzed HCV clones of the hypervariable region in four HCV RNA-positive infants and compared them with those of the mother. Cord blood samples from three of the four infants were positive for the HCV RNA (< or =10(3) copies/mL), and all of the four infants had the HCV RNA titer of >10(6) copies/mL within 2 mo after birth. The hypervariable region clones detected in the infants were closely related to those in the respective mothers. The results suggest the perinatal transmission of HCV. The hypervariable region clones transmitted to infants were not a single selected clone or minor clones in the mother. None of the clones specific to the low-density fraction in the mother was transmitted to the infants. Moreover, the proportion of HCV in the low-density fraction was minimal in the first few months of life, but increased several months after birth in association with the elevation of alanine aminotransferase. These results suggest that the increase of HCV in the low density fraction reflect the evolution of immune response in infants. We also demonstrated that the emergence of quasispecies in infants precedes the infantile antibody response. PMID- 11004235 TI - Does low IgA in human milk predispose the infant to development of cow's milk allergy? AB - We sought a relationship between total and cow's milk-specific IgA levels in colostrum and human milk and subsequent development of cow's milk allergy (CMA) in the breast-fed infant. The study included 87 nursing mothers and their infants (age, 2 d to 7 mo), followed prospectively up to 1 y. At 1 y, 48 mothers (69% with an atopic constitution) had an infant with CMA, verified by clinical cow's milk challenge, eight (38% with an atopic constitution) had a baby who had had protracted infantile colic but no CMA (disease control group), and 31 (23% with an atopic constitution) had a healthy infant. Total breast-milk IgA was measured by radial immunodiffusion, and IgA antibodies to cow's milk were measured by ELISA during the breast-feeding period. The levels of total and cow's milk specific IgA antibodies in colostrum and human milk were significantly lower in the mothers whose baby later developed CMA [estimated third day value, 0.38 g/L (95% confidence interval, 0. 24-0.82)] than in the ones whose infant remained healthy or had had infantile colic but not CMA [0.82 g/L (95% confidence interval, 0. 99-1.51); p < 0.05]. The infants developed CMA significantly more often if the concentration of total IgA antibodies in milk was <0.25 g/L, when measured between 6 d and 4 wk postpartum [sensitivity, 0. 55; specificity, 0.92; odds ratio, 14.7 (95% confidence interval, 3. 1-70.2); p < 0.001]. The levels of cow's milk-specific IgA positively correlated with the levels of total IgA but not with the development of CMA in the infant. The levels of total or cow's milk specific IgA did not correlate with maternal atopy. IgA antibodies in colostrum and human milk may prevent antigen entry at the intestinal surface of the breast fed infant. A low IgA content in human milk may lead to defective exclusion of food antigens and thus predispose an offspring to develop food allergies. PMID- 11004236 TI - Inv(11)(p13p15) and myf-3(MyoD1) in a malignant extrarenal rhabdoid tumor of a premature newborn. AB - We present a premature newborn of 32 wk of gestation with a congenital malignant extrarenal rhabdoid tumor (MERT) on the right shoulder with generalized metastases. Standard histologic, immunohistochemical, molecular and cytogenetic methods were used in the evaluation of diagnostic material. Biopsy of a skin lesion showed the histologic features of a malignant rhabdoid tumor. Cytogenetic analysis of the tumor cells revealed an inv(11)(p13p15) and additionally, an increased expression of myf-3 (myogenic determination factor, MyoD1) and PAX3 was detected. These results suggest an origin of the neoplasm derived from a pluripotent cell with the potential of myogenic differentiation. Tumor suppressor genes located on chromosome 11p13 and 11p15 may play an important role for malignant rhabdoid tumor development and progression. PMID- 11004237 TI - Comparative analysis of different puberty inhibiting mechanisms of two GnRH agonists and the GnRH antagonist cetrorelix using a female rat model. AB - GnRH agonists are the established treatment of precocious puberty caused by premature stimulation of gonadotropin secretion. It has been reported that after an initial stimulation ("flare-up") they reduce LH secretion by desensitization of pituitary GnRH receptors. Little has been published about the use of GnRH antagonists such as cetrorelix to control the onset of puberty and whether they are potentially advantageous compared with GnRH agonists. We conducted two multigroup experiments (12 and 10 d, respectively) treating prepubertal/peripubertal female rats with either the GnRH agonist triptorelin or buserelin and compared them with rats treated with the GnRH antagonist cetrorelix and controls to assess the effects on pubertal progress and serum hormones. In the second experiment, the effects of buserelin and cetrorelix on gene expression of the GnRH receptor, LH-beta, FSH-beta, and the alpha subunit genes in the pituitary were also investigated. Cetrorelix, triptorelin, and buserelin retarded the onset of puberty as determined by delayed vaginal opening, lower ovarian weights, and lower serum estradiol levels. However, although LH and FSH levels were stimulated by both agonists, they were inhibited by cetrorelix. In the cetrorelix versus buserelin experiment, pituitary gene expression of the GnRH receptor and LH-beta subunit were significantly lower in cetrorelix treated rats compared with controls whereas buserelin had little effect. Expression of FSH beta and alpha subunit were stimulated by buserelin but not by cetrorelix. Even though all three of these GnRH analogues inhibited gonadal development and delayed the onset of puberty, the GnRH agonists had stimulating and inhibiting effects on the pituitary-gonadal axis whereas cetrorelix exerted only inhibiting effects. We conclude from this female rat model that cetrorelix may offer advantages for a more controlled medical treatment of precocious puberty compared with GnRH agonist treatment. PMID- 11004238 TI - Validated multivariate models predicting the growth response to GH treatment in individual short children with a broad range in GH secretion capacities. AB - The aim of the study was to develop and validate models that could predict the growth responses to GH therapy of individual children. Models for prediction of the initial one and 2-y growth response were constructed from a cohort of 269 prepubertal children (Model group) with isolated GH deficiency or idiopathic short stature, using a nonlinear multivariate data fitting technique. Five sets of clinical information were used. The "Basic model" was created using auxological data from the year before the start of GH treatment and parental heights. In addition to Basic model data, the other four models included growth data from the first 2 y of life, or IGF-I, or GH secretion estimated during a provocation test (AITT) or a spontaneous GH secretion profile. The performance of the models was validated by calculating the differences between predicted and observed growth responses in 149 new GH treated children (Validation group) who fulfilled the inclusion criteria used in the original cohort. The SD of these differences (SD(res)) in the validation group was compared with the SD(res) for the model group. For the 1st y, the SD(res) for the Basic model was 0.28 SDscores. The lowest SD(res) (0.19 SDscores), giving the most narrow prediction interval, was achieved adding the 24h GH profile and data on growth from the first 2 y of life to the Basic model. The models presented permit estimation of GH responsiveness in children over a broad range in GH secretion, and with an accuracy of the models substantially better than when using maximal GH response during an provocation test. The predicted individual growth response, calculated using a computer program, can serve as a guide for evidence-based decisions when selecting children to GH treatment. PMID- 11004239 TI - Physiologic predictors of lumbar spine bone mass in neonates. AB - Dual energy x-ray absorptiometry (DXA) of the lumbar spine (LS) was measured in 201 singleton infants with birth weights from 1152 to 3970 g and gestational ages from 27 to 42 wk. All infants were well and studied at a mean (+/-SD) of 2.1 (+/ 1.6) days after birth. There were 75 Caucasian (46 males, 29 females) and 126 African American infants (58 males, 68 females). Scan acquisition of the first to fourth lumbar vertebrae was performed with a single beam whole body scanner (Hologic QDR 1000/W densitometer, Hologic Inc, Waltham, MA, U.S.A.) using the infant spine mode. Scan analysis was performed with software version 4.57Q and consistent region of interest. The SD of difference for duplicate LS scans is <1.4% at a mean bone mineral content (BMC) of 2.14 g. Results show that LS BMC, area, bone mineral density (BMD) increased by approximately 550%, 280% and 180%, respectively, between 27 and 42 wk gestational age. Body mass accounted for about 70% and 55% of the variance in BMC and BMD respectively. In contrast, the infant's length appears to be the best determinant of LS area and accounts for about 75% of the variance in LS area. Race, gender or season has little or no effect on LS bone mass. There was progressive increase in BMC and area from first to fourth lumbar vertebra but BMD was significantly higher only at the fourth lumbar vertebra. We conclude that DXA LS can be performed even in small preterm infants. Its excellent precision, low radiation exposure and rapid scan acquisition offers promise as a useful tool for widespread use in pediatrics. Our data may be used as a basis for further studies in physiologic and pathologic situations that may affect bone mineralization in infants. PMID- 11004240 TI - Bone turnover and growth during and after continuing chemotherapy in children with acute lymphoblastic leukemia. AB - Children treated for acute lymphoblastic leukemia may develop reduced bone mineral density during treatment, but there is little information on the mechanisms involved. In a prospective, longitudinal study on 15 children with ALL, we undertook serial measurements of markers of bone and collagen turnover, insulin-like growth factor (IGF)-I and its binding proteins (IGFBPs)-3 and -2 during the second year of continuing chemotherapy. In eight patients we also measured lower leg length by knemometry. Height SD scores, lower leg length velocity, IGF-I, and markers of bone collagen turnover did not differ significantly from healthy children. However, bone alkaline phosphatase, a marker of the differentiated osteoblast, was lower (mean SD score, -0.64; p < 0.0001), whereas procollagen type III N-terminal propeptide (P3NP, a marker of soft tissue collagen turnover; mean SD score, +0.93, p < 0.05), IGFBP-3 (mean SD score, +0.76; p < 0.01), and IGFBP-2 (mean SD score, +1.24, p = 0.01) were all higher than in healthy children. IGFBP-3 decreased during episodes of afebrile neutropenia (p < 0.05). Within 3 mo after completion of treatment, bone ALP increased in all eight patients, but collagen markers showed little change. IGFBP 2 returned to normal posttreatment, but P3NP and IGFBP-3 remained significantly elevated compared with healthy children (mean SD scores, +1.51 and +1.36, respectively; p < 0.01). We conclude that continuing chemotherapy was associated with normal growth and bone collagen turnover but enhanced soft tissue collagen turnover. Bone bone alkaline phosphatase was low throughout treatment, which suggests impaired osteoblast differentiation resulting from a direct effect of chemotherapy on bone. Although the effect was reversible, the long-term implications for bone health in survivors remain uncertain. PMID- 11004241 TI - Investigation of three doses of oral insulin-like growth factor-I on jejunal lactase phlorizin hydrolase activity and gene expression and enterocyte proliferation and migration in piglets. AB - In a previous study, oral IGF-I at 65 nM increased lactase phlorizin hydrolase (LPH) activity and villus height in piglets, however, the mechanisms were unknown. Herein, the response to a range of doses of IGF-I was investigated and we hypothesized that LPH and villus height would respond to oral IGF-I in a dose dependent manner. Two 14-d experiments were conducted using cesarean-derived piglets. In experiment 1, piglets (n = 28) were fed formula containing 0, 33, 65, or 131 nmol/L (0, 0.25, 0.5, or 1.0 mg/L) recombinant human IGF-I. In experiment 2, 5'-bromodeoxyuridine was administered to piglets fed formula alone (n = 4) or containing 131 nmol/L IGF-I (n = 4). IGF-I did not affect body weight gain or intestinal weight or length. Jejunal villus height and LPH activity were significantly greater in piglets fed 131 nmol IGF-I/L than control piglets. Villus height and lactase activity in piglets fed the 33 and 65 nmol/L IGF-I doses were similar and intermediate between control and 131 nmol IGF-I/L. Jejunal mRNA expression and LPH polypeptide abundance were investigated in piglets receiving 0 or 131 nmol/L IGF-I. Steady state LPH mRNA abundance was significantly higher (p < 0.05) in IGF-I-treated piglets. The relative abundance of proLPH(h) was not significantly increased (p = 0.06) by IGF-I treatment. Mucosal DNA content and DNA synthesis were greater in piglets receiving 131 nmol IGF-I/L than control, however, enterocyte migration and mucosal protein content were unaffected. Thus, oral IGF-I increased jejunal LPH activity and LPH mRNA abundance and stimulated intestinal cell hyperplasia in normal piglets. PMID- 11004242 TI - Functional characterization of the keratinocyte growth factor system in human fetal gastrointestinal tract. AB - Keratinocyte growth factor (KGF) is a paracrine growth factor whose mRNA has been detected in human adult and rodent gut tissues together with its associated receptor. Our objectives were to assess the presence of immunoreactive KGF ligand and receptor proteins in human fetal gastrointestinal (GI) tract segments and to evaluate the role of exogenous KGF on cell proliferation and intestinal digestive functions. KGF (26-28 kD doublet) was identified in esophagus, stomach, small intestine, and colon by Western blot. Its receptor (135 kD) was ubiquitously detected in proliferative and differentiated epithelial cells of each GI segment by use of indirect immunofluorescence (anti-bek, anti-K-sam). The addition of KGF to explants cultured in serum-free conditions greatly stimulated DNA synthesis in all GI tract tissues. The growth factor up-regulated intestinal sucrase isomaltase and gamma-glutamyl-transpeptidase activities in jejunal explants, whereas it down-regulated these activities in colon explants. It is suggested that the KGF system likely represents an important paracrine pathway that is able to stimulate cell proliferation in all segments of the human fetal GI tract and to differentially regulate intestinal digestive functions. PMID- 11004243 TI - Nonnutritive factors in colostrum enhance myofibrillar protein synthesis in the newborn Pig. AB - Colostrum is a complex source of nutrients, immune factors, and bioactive substances consumed by newborn mammals. In previous work, we observed that protein synthesis in the skeletal muscle of newborn piglets is enhanced when they are fed colostrum rather than a nutrient-matched formula devoid of growth factors. To elucidate the mechanisms responsible for this response, we contrasted the fractional rates of sarcoplasmic and myofibrillar protein synthesis of newborn piglets that received only water with those fed for 24 h with colostrum, a nutrient-matched formula, or mature sow's milk. Compared with water, feeding resulted in a 2.5- to 3-fold increase in total skeletal muscle protein synthesis, and this increase was 28% greater in the colostrum-fed than either the formula- or mature milk-fed piglets. Feeding also stimulated muscle ribosome and total polyadenylated RNA accretion. Ribosomal translational efficiency, however, was similar across all fed groups. The greater stimulation of protein synthesis in colostrum-fed pigs was restricted entirely to the myofibrillar protein compartment and was associated with higher ribosome and myosin heavy chain mRNA abundance. Taken together, these data suggest that nonnutritive factors in colostrum enhance ribosomal accretion and muscle-specific gene transcription that, in turn, stimulate specifically the synthesis of myofibrillar proteins in the skeletal musculature of the newborn. PMID- 11004244 TI - Factors affecting the measurement of energy expenditure during energy balance studies in preterm infants. AB - Our objective was to examine factors that affect the accuracy of energy expenditure measurements (EE), when using flow-through indirect calorimetry (IC), to determine the minimum length of time needed to measure 24- and 48-h EE and to compare cross-over and parallel designs as methods of investigation during energy balance collections (EB) in preterm infants. A baby doll manikin was used to determine equilibration times and to compare VCO(2) and VO(2) as measured by flow meter and indirect calorimetry under different study conditions, one of which simulated an EB. "Continuous" EE was measured to determine the minimum length of time needed to accurately reflect 24- and 48-h EE and to compare parallel and cross-over studies as methods of study design in a group of "normal" enterally fed preterm infants. The mean (+/-SD) errors between flow meter and indirect calorimetry determinations for VCO(2) and VO(2) were -1.9 +/- 2.5 and -1.8 +/- 4.3% under conditions that simulated an EB. Cumulative 6-h EE accurately predicted 24- and 48-h EE. Expressed in absolute terms (kcal/d), EE did not change on a day-to-day basis but did increase over the 2-wk study period. Expressed on body weight basis (kcal/kg/d), EE did not change on a day-to-day or week-to-week basis. The variance in EE due to biologic variability; i.e. the parallel design, was approximately 6 times greater than that due to age, weight, and weight gain; i.e. the cross-over design. Indirect calorimetry, therefore, accurately measures EE in conditions simulating an energy balance collection. Six hour EE determinations are valid estimate of EE during a 48-h balance collection, while cross-over studies may be the preferred method of study design during short term studies of EE in preterm infants. PMID- 11004245 TI - Dietary 20:4n-6 and 22:6n-3 modulates the profile of long- and very-long-chain fatty acids, rhodopsin content, and kinetics in developing photoreceptor cells. AB - The objective of this study was to determine whether addition of dietary 20:4n-6 and 22:6n-3 to a conventional infant formula fat blend influences membrane long chain and very-long-chain fatty acid composition, rhodopsin content, and rhodopsin kinetics in developing rat photoreceptor cells. The dietary fats were formulated based on the fat composition of a conventional infant formula providing an 18:2n-6/18:3n-3 ratio of 7:1 (SMA, Wyeth Nutritionals), which served as the control fat blend. This dietary fat blend was modified to contain 20:4n-6 [arachidonic acid (AA)], 22:6n-3 [docosahexaenoic acid (DHA)], AA + DHA, or an 18:2n-6/18:3n-3 ratio of 4:1 (alpha-linolenic acid). Dams were fed diets from birth, and rat pups were fed the same diet after weaning. Retinas and rod outer segments were prepared in the dark from pups at 2, 3, and 6 wk of age for fatty acid analysis of individual phospholipids, rhodopsin content, and rhodopsin disappearance kinetics after light exposure. Feeding AA + DHA in the diet increased 22:6n-3 levels in phosphatidylcholine and phosphatidylethanolamine. In phosphatidylcholine, total n-6 tetraenoic very-long-chain fatty acids and total n 3 pentaenoic and n-3 hexaenoic very-long-chain fatty acids increased after feeding AA and DHA, respectively. Developmental changes were characterized by a decrease in 20:4n-6 in the major phospholipids, whereas 22:6n-3 increased with age in rod outer segments. The highest rhodopsin content occurred in the retina of rats fed diets containing AA and/or DHA. The kinetics of rhodopsin disappearance after light exposure was highest in rats fed DHA at 6 wk of age. This study demonstrates that small manipulations of the dietary level of 20:4n-6 and 22:6n-3 are important determinants of fatty acid composition of membrane lipid and visual pigment content and kinetics in the developing photoreceptor cell. PMID- 11004246 TI - Lipopolysaccharides do not alter metabolic disturbances in hippocampal slices of fetal guinea pigs after oxygen-glucose deprivation. AB - The aim of the present study was to clarify whether endotoxins [lipopolysaccharides (LPS)] have a toxic effect on fetal brain tissue after cerebral ischemia, while excluding their effect on the cardiovascular system. Experiments were therefore performed on hippocampal slices prepared from mature fetal guinea pigs. In particular, we studied the influence of LPS on nitric oxide production, energy metabolism, and protein synthesis after oxygen-glucose deprivation (OGD). Incubating hippocampal slices in LPS (4 mg/L) for as long as 12 h did not alter cGMP tissue concentrations significantly. However, 10 min after OGD of 40-min duration, cGMP tissue concentrations were substantially increased in relation to controls, and this increase was almost completely blocked by the application of 100 microM N:(omega)-nitro-L-arginine, indicating that nitric oxide synthase was activated after OGD in fetal brain tissue. Again, LPS did not have any effect on cGMP tissue concentrations after OGD. Furthermore, addition of LPS altered neither protein synthesis nor energy metabolism measured 12 h after OGD. We therefore conclude that, apart from their well-known influence on the cardiovascular system, LPS do not alter metabolic disturbances in hippocampal slices of fetal guinea pigs 12 h after OGD. A direct toxic effect of LPS on immature brain tissue within this interval does not therefore seem to be very likely. However, delayed activation of LPS-sensitive pathways that may be involved in cell death, or damage limited to a small subgroup of cells such as oligodendrocyte progenitors, cannot be fully excluded. PMID- 11004247 TI - Three novel PHEX gene mutations in Japanese patients with X-linked hypophosphatemic rickets. AB - X-linked hypophosphatemic rickets (XLH) is an X-linked dominant disorder characterized by renal phosphate wasting, abnormal vitamin D metabolism, and defects of bone mineralization. The phosphate-regulating gene on the X-chromosome (PHEX) that is defective in XLH has been cloned, and its location identified at Xp22.1. It has been recognized to be homologous to certain endopeptidases. So far, a variety of PHEX mutations have been identified mainly in European and North American patients with XLH. To analyze the molecular basis of four unrelated Japanese families with XLH, we determined the nucleotide sequence of the PHEX gene of affected members. We detected a new nonsense mutation (R198X) in exon 5, a new 3 nucleotides insertion mutation in exon 12 and a new missense mutation (L160R) in exon 5 as well as a previously reported nonsense mutation in exon 8 (R291X). These results suggest that: 1) PHEX gene mutations are responsible for XLH in Japanese patients, and 2) PHEX gene mutations are heterogeneous in the Japanese population similarly to other ethnic populations. PMID- 11004248 TI - Temperature-sensitive mutation of PEX6 in peroxisome biogenesis disorders in complementation group C (CG-C): comparative study of PEX6 and PEX1. AB - Peroxisome biogenesis disorders (PBD), including Zellweger syndrome, neonatal adrenoleukodystrophy, and infantile Refsum disease, are a group of genetically heterogeneous autosomal-recessive diseases caused by mutations in PEX genes that encode peroxins, proteins required for peroxisome biogenesis. Zellweger syndrome patients present the most severe phenotype, whereas neonatal adrenoleukodystrophy patients are intermediate and infantile Refsum disease patients have the mildest features. PEX6 is a causative gene for PBD of complementation group C (CG-C) and encodes the peroxin Pex6p, one of the ATPases associated with diverse cellular activities and a member of the same family of proteins as Pex1p, a causative protein for PBD of CG-E (CG1). Here, we identified the temperature sensitivity of peroxisomes in the fibroblasts of a patient with neonatal adrenoleukodystrophy in CG-C. Peroxisomes were morphologically and biochemically formed at 30 degrees C but not at 37 degrees C. This patient was homozygous for a missense mutation, T- >C at nucleotide 170 resulting in a change from leucine to proline at amino acid 57 (L57P) in Pex6p. CG-C cell mutants (ZP92) in the Chinese hamster ovary transfected with L57P in HsPEX6 revealed the same temperature-sensitive phenotype. However, PEX1-deficient Chinese hamster ovary cell mutants (ZP101) transfected with L111P in PEX1, the counterpart to L57P in PEX6, showed no temperature sensitivity. In addition, ZP92 transfected with G708D in PEX6, the counterpart to the temperature-sensitive mutation G843D in PEX1, revealed no temperature-sensitive phenotype. These results indicate that L57P in Pex6p is a temperature-sensitive mutation causing the milder phenotype in a patient with PBD in CG-C. They also indicate that the amino acid residues responsible for temperature sensitivity do not seem to be conserved between Pex6p and Pex1p. PMID- 11004250 TI - Lung water and proton magnetic resonance relaxation in preterm and term rabbit pups: their relation to tissue hyaluronan. AB - The present study was performed to investigate simultaneously total lung water, T(1) and T(2) relaxation times, and hyaluronan (HA) in preterm and term rabbits. Attempts were also made to establish the relationship of HA to total lung water and to T(2)-derived motionally distinct water fractions. Experiments were performed in fetal Pannon white rabbit pups at gestational ages of 25, 27, 29, and 31 d and at a postnatal age of 4 d. Lung tissue water content (desiccation method), T(1) and T(2) relaxation times (H(1)-NMR method), and HA concentration (radioassay) were measured, and free and bound water fractions were calculated by using multicomponent fits of the T(2) relaxation curves. Lung water content and T(1) and T(2) relaxation times were highest at a gestational age of 27 d and then declined steadily during the whole study period. Similar trends and time courses were seen for the fast and slow components of the T(2) relaxation curve. The T(2) derived free water fraction remained unchanged at a gestational age of 25-29 d ( approximately 67%), but increased progressively to a value of 78.5 +/- 7.9% at 31 d (p < 0. 001) and to 83.4 +/- 9.4% at the postnatal age of 4 d (p < 0.01). Opposite changes occurred in the bound water fraction. Lung HA concentration decreased with advancing gestation from 870.8 +/- 205.2 microg/g dry weight at 25 d to 162.6 +/- 32.4 microg/g dry weight at 31 d (p < 0.001), but it was increased 2-fold postnatally. HA correlated positively with total lung water (r = 0.39; p < 0.001) but not with the bound water fraction. It is suggested that the physiologic lung dehydration is associated with macromolecule-related reorganization of lung water and that the role of HA in this process needs to be further investigated. PMID- 11004249 TI - Relaxant effects of carbon monoxide compared with nitric oxide in pulmonary and systemic vessels of newborn piglets. AB - Nitric oxide (NO) has been implicated in a number of diverse physiologic processes, including regulation of vascular tone. Carbon monoxide (CO) is another endogenously generated diatomic gas that may play an important physiologic role in vascular smooth muscle homeostasis. The purpose of this study was to compare the responses to exogenous NO and CO in isolated vessels (pulmonary arteries, pulmonary veins, and mesenteric arteries) from 12- to 24-h-old and 2-wk-old piglets. Vessels precontracted with the thromboxane A(2) mimetic U46619 (10(-7) M) relaxed in response to CO (2 x 10(-6) to 2 x 10(-4) M) and NO (2 x 10(-9) to 2 x 10(-7) M); these effects were not affected by endothelium removal but were completely abolished by the soluble guanylate cyclase inhibitor ODQ (10(-5) M). In pulmonary arteries, the maximal relaxation to NO increased with postnatal age from 33 +/- 4% of the precontraction value to 56 +/- 5%, in 12- to 24-h-old and 2 week-old piglets, respectively (p < 0.01), but the response to CO decreased from 25 +/- 3% to 12 +/- 1%, respectively (p < 0.01). The maximal response to CO was greater in pulmonary veins than in pulmonary or mesenteric arteries for both age groups (p < 0.01). Vasorelaxation induced by endogenous NO (stimulated by acetylcholine) was also greater in pulmonary veins when compared with pulmonary arteries and increased with postnatal age in both vessels. In contrast, no age related differences were observed in the vasorelaxation induced by the cGMP analog 8-bromo cGMP in pulmonary arteries. When the response to NO was analyzed under three different extracellular O(2) concentrations (PO(2) 4.51 +/- 0.03, 19. 32 +/- 0.17, and 86 +/- 0.62, kPa), no significant differences were found. However, in the presence of superoxide dismutase (100 U/mL). the response to CO remained unchanged, and the response to NO improved in pulmonary arteries from 2 week-old but not from newborn piglets. In conclusion, both NO and CO relaxed neonatal vessels through soluble guanylate cyclase activation. However, when compared with NO, CO exhibited a poor vasorelaxant activity. Pulmonary vasorelaxation induced by NO increased with postnatal age, whereas that induced by CO decreased. Changes in extracellular oxygen concentration did not alter the pulmonary vascular response to NO. However, the presence of superoxide dismutase improved the response to NO, indicating that oxidant activity limits the vasorelaxant response to NO but not to CO. PMID- 11004251 TI - Effects of deferoxamine, a chelator of free iron, on NA(+), K(+)-ATPase activity of cortical brain cell membrane during early reperfusion after hypoxia-ischemia in newborn lambs. AB - Free iron chelation after hypoxia-ischemia can reduce free radical-induced damage to brain cell membranes and preserve electrical brain activity. We investigated whether chelation of free iron with deferoxamine (DFO) preserved cortical cell membrane activity of Na(+),K(+)-ATPase and electrocortical brain activity (ECBA) of newborn lambs during early reperfusion after severe hypoxia-ischemia. Hypoxia was induced in 16 lambs by decreasing the fraction of inspired oxygen to 0.07 for 30 min, followed by a 5-min period of hypotension (mean arterial blood pressure <35 mm Hg). ECBA (in microvolts) was measured using a cerebral function monitor. Immediately after hypoxia and additional ischemia, eight lambs received DFO (2.5 mg/kg, i.v.), and seven lambs received a placebo (PLAC). Two lambs underwent sham operation. One hundred eighty minutes after completion of hypoxia and ischemia, the brains were obtained and frozen. Na(+),K(+)-ATPase activity was measured in the P(2) fraction of cortical tissue. Na(+),K(+)-ATPase activity was 35.1 +/- 7.4, 42.0 +/- 7.6, and 40.7 +/- 1.4 micromol inorganic phosphate/mg protein per hour in PLAC-treated, DFO-treated, and sham-operated lambs, respectively (p < 0.05: DFO versus PLAC). ECBA was 11.2 +/- 6.1, 14.8 +/- 4.8, and 17.5+/-.0.5 microV in PLAC-treated, DFO-treated, and sham-operated lambs, respectively (p = 0.06: DFO versus PLAC). Na(+),K(+)-ATPase activity correlated with ECBA at 180 min of reperfusion (r = 0.85, p < 0.001). We conclude that Na(+),K(+)-ATPase activity of cortical brain tissue was higher in DFO-treated lambs compared with PLAC-treated animals during the early reperfusion phase after severe hypoxia ischemia, suggesting a reduction of free radical formation by DFO. Furthermore, a positive relationship was found between Na(+),K(+)-ATPase activity and ECBA. PMID- 11004252 TI - Esterification of vitamin A by the human placenta involves villous mesenchymal fibroblasts. AB - Vitamin A (retinol) and its active derivatives (retinoic acids) are essential for growth and development of the mammalian fetus. Maternally derived retinol must pass the placenta to reach the developing fetus. Despite its apparent importance, little is known concerning placental transfer and metabolism of retinol, and particularly of placental production and storage of retinyl esters. To elucidate this metabolic pathway, we incubated, in the presence of retinol, 1) human full term placental explants and 2) primary cultures of major cells types contributing to placental function: trophoblasts and villous mesenchymal fibroblasts. We used HPLC to determine the types and concentrations of retinyl esters produced by these explants and cells. About 14% of total cellular retinol in placental explants was esterified. The most abundant esters were myristate and palmitate. Primary cell cultures showed that fibroblasts efficiently produced retinyl esters, but trophoblasts did not. In both types of experiments, no retinyl esters were detected in the culture medium, suggesting that retinyl esters were produced for storage purpose. These results suggest that villous mesenchymal fibroblasts are primary sites of retinol esterification and storage in the placenta. PMID- 11004253 TI - Comparison of visual results in initial and re-treatment laser in situ keratomileusis procedures for myopia and astigmatism. AB - PURPOSE: To report the results of initial and re-treatment laser in situ keratomileusis procedures for myopia and astigmatism. METHODS: A total of 1161 consecutive inital and retreatment laser in situ keratomileusis procedures in 697 patients performed between September 19, 1996, and June 20, 1998, were retrospectively analyzed. Baseline and postoperative best spectacle-corrected visual acuity, uncorrected visual acuity, spherical and cylindrical refractions, computerized video keratography, and biomicroscopy were measured or performed on each eye. RESULTS: Of 1,071 eyes, 900 (84%) underwent a single-laser in situ keratomileusis procedure, and 171 of 1,071 eyes (16%) underwent one or two re treatment procedures. The preoperative mean +/- SD spherical equivalent was -5.09 +/- 3.15 diopters (range, -0.75 to -14.38 diopters) and the cylinder was 0.97 +/- 1.09 diopters (range, 0.00 to 3.25 diopters) in the group that underwent a single laser in situ keratomileusis treatment. The preoperative mean +/- SD spherical equivalent was -6.26 +/- 3.04 diopters (range, -1.75 to -12.88 diopters) and the cylinder was 1.53 +/- 1.11 diopters (range, 0.00 to 3.50 diopters) in the group that underwent re-treatment. Before re-treatment, 110 eyes (64.3%) in the group that underwent re-treatment achieved 20/40 or better uncorrected visual acuity and 23 eyes (13.5%) achieved 20/25 or better. Three months after re-treatment, 155 eyes (90.6%) achieved 20/40 or better, and 80 eyes (46.8%) achieved 20/25 or better. In the single-procedure group, uncorrected visual acuity at 3 months revealed 20/40 vision or better in 856 eyes (95.1%) and 20/25 vision or better in 572 eyes (63.6%). In this study group, 23.6% of eyes with a preoperative spherical equivalent greater than -6.0 diopters underwent re-treatment compared with 11.8% of eyes with a preoperative spherical equivalent of -6.0 diopters or less. CONCLUSIONS: Laser in situ keratomileusis appears to be an effective procedure for mild, moderate, and severe myopia. In eyes with residual refractive error, re-treatment laser in situ keratomileusis procedures can result in good visual outcomes. PMID- 11004254 TI - Photorefractive keratectomy for compound myopic astigmatism. AB - PURPOSE: To evaluate the safety and efficacy of photorefractive keratectomy for the treatment of primary compound myopic astigmatism. METHODS: In a prospective study, 93 eyes from 56 patients with a mean spherical equivalent of -4.98 +/- 1.80 diopters (range, -1.75 to -8.5) underwent photoastigmatic refractive keratectomy with the Summit Apex Plus excimer laser using erodible mask technology and were followed for 2 years. Primary outcome measures included an assessment of astigmatic correction through vector analysis, manifest refraction, uncorrected visual acuity, corneal clarity, and the presence of adverse symptoms. RESULTS: Eighty-five eyes (91.4%) were available for analysis at 6 months. Mean spherical equivalent refraction was reduced 85% (mean, -0.75 +/- 0.85 diopter) and the target-induced astigmatism was reduced 70% (mean, 0.98 +/- 1.88 diopters). Forty-eight eyes (56%) had an uncorrected visual acuity of 20/20 or greater, whereas 70 eyes (82%) had an uncorrected visual acuity of 20/40 or greater. Twenty-four eyes (26% ) required re-treatment because of undercorrection of the spherical equivalent and astigmatic components after the 6-month follow up. Fifty-nine of the remaining eyes were available at the 24-month visit. Mean spherical equivalent refraction was reduced to -0.39 +/- 0.72 diopter (91.8%). The target-induced astigmatism was reduced 64% from 1.74 diopters. Forty-one eyes (81.3%) were within +/-1.0 diopter of attempted spherical equivalent correction. Stability within a spherical equivalent of +/-0.5 diopter occurred after the first postoperative month. Fifty-six eyes (94.9%) had an uncorrected visual acuity of 20/40 or greater, whereas 34 eyes (57.6 %) demonstrated an uncorrected visual acuity of 20/20 or greater. One eye (1.7%) lost 2 or more lines of best spectacle-corrected visual acuity. CONCLUSION: Photoastigmatic refractive keratectomy with the Summit Apex Plus excimer laser is a safe and effective method of reducing compound myopic astigmatism. However, higher re-treatment rates may result from significant undercorrections because of current laser algorithms and variability in the mean angle of error. PMID- 11004255 TI - The effectiveness of a topical antibiotic irrigating solution in a model of staphylococcal keratitis after lamellar keratectomy. AB - PURPOSE: To create a model of Staphylococcus aureus keratitis after lamellar keratectomy; to assess the toxicity of an antibiotic irrigating solution on the corneal stromal bed; and to test the chemotherapeutic effectiveness of a topical antibiotic, both alone and with an antibiotic-containing irrigating solution in preventing S. aureus keratitis after lamellar keratectomy. METHODS: The right eye of each of 38 rabbits were used in this study. In 18 eyes, a lamellar flap was created with a microkeratome, and an inoculum of S. aureus (either 1,000, 5,000, or 50,000 CFUs) was instilled under each flap; the eyes were examined for signs of infection and inflammation at 24 and 48 hours. In another five eyes, a lamellar flap was created in the same manner and the stromal bed was irrigated with 0.3% ofloxacin; the eyes were assessed for ocular inflammatory changes and evidence of crystalline deposits. Finally, in each of 15 additional eyes, 1,000 CFUs of S. aureus were instilled under a lamellar flap to create experimental infectious keratitis. The keratitis was treated according to three regimens: irrigation of the stromal bed with sterile balanced salt solution; irrigation of the stromal bed with 0.3% ofloxacin, followed by application of topical ofloxacin four times a day; application of topical ofloxacin only, four times a day. Eyes were examined for infection and ocular inflammatory changes at 24 and 48 hours. RESULTS: Staphylococcus aureus keratitis can consistently be produced under the stromal flap by inoculation of relatively few organisms. Irrigation of the stromal bed with commercial-strength topical ofloxacin does not appear to be toxic to the stromal bed, with no evidence of crystalline precipitates of the antibiotic. In our model of infectious keratitis after lamellar keratectomy, both topical ofloxacin alone and the combination of topical ofloxacin and irrigation of the stromal bed with 0.3% ofloxacin were effective at preventing S. aureus keratitis. However, the combined treatment of antibiotic irrigation plus topical antibiotic was more effective at preventing inflammation than topical ofloxacin alone. CONCLUSIONS: In this model of S. aureus keratitis after lamellar keratectomy, irrigation of the stromal bed with antibiotic plus topical antibiotic appears to be both safe and effective for preventing infection. PMID- 11004256 TI - Gas-permeable scleral contact lens therapy in ocular surface disease. AB - PURPOSE: To describe the therapeutic benefits of nonfenestrated gas-permeable scleral contact lenses in the management of patients with ocular surface disease. METHODS: The charts of 49 consecutive patients (76 eyes) with ocular surface disease whose management included the use of gas-permeable scleral contact lenses were reviewed. We also developed a questionnaire to assess the impact of lens wear on subjective aspects of activities of daily living. RESULTS: The mean age of the 49 patients was 44.6 years (range, 3 to 87 years); 31 patients were female and 18 were male. The most common indication for fitting of the lenses was Stevens-Johnson syndrome (54 [71%] of the 76 eyes). Other indications included ocular cicatricial pemphigoid, exposure keratitis, toxic epidermal necrolysis, postherpetic keratitis, congenital deficiency of meibomian glands, superior limbal keratoconjunctivitis, Sjogren syndrome, and inflammatory corneal degeneration. The mean follow-up was 33.6 months (range, 2 to 144 months). Improvement in best-corrected visual acuity (defined as a gain of 2 or more Snellen lines) was observed in 40 (53%) of the eyes. In eight (53%) of the 15 eyes with active corneal epithelial defects at the time of lens fitting, the defects healed, whereas in the remaining seven eyes the corneal epithelial defects remained unchanged. Forty-five (92%) of the 49 patients reported improvement in their quality of life as a result of reduction of photophobia and discomfort. The mean wearing time of the gas-permeable scleral contact lenses was 13.7 hours per day (range, 4 to 18 hours). Many patients had preparatory surgical procedures before lens fitting (for example, punctal occlusion or mucous membrane grafting), and some had visual rehabilitation surgical procedures (for example, keratoplasty and/or cataract surgery) after lens fitting. CONCLUSIONS: Gas permeable scleral contact lens wear provides an additional effective strategy in the surface management and visual rehabilitation of patients with severe ocular surface disease. PMID- 11004257 TI - Treatment of persistent corneal epithelial defect with extended wear of a fluid ventilated gas-permeable scleral contact lens. AB - PURPOSE: To report treatment of persistent corneal epithelial defects unresponsive to other therapies by extended wear of a fluid-ventilated gas permeable scleral contact lens. METHODS: In this retrospective study, 14 eyes of 13 consecutive patients referred for the treatment of persistent corneal epithelial defects that failed to heal with conventional therapies or developed epithelial defects after penetrating keratoplasty for persistent corneal epithelial defects were fitted with an extended-wear gas-permeable scleral lens. These included seven eyes of six patients with Stevens-Johnson syndrome and seven eyes of seven patients who did not have Stevens-Johnson syndrome. Twelve eyes had undergone recent penetrating keratoplasty. All 14 eyes were fitted with a gas permeable scleral contact lens designed to avoid the intrusion of air bubbles under its optic. An antibiotic and corticosteroid were added to the lens fluid reservoir or instilled before each lens insertion in 12 of 14 eyes. The lenses were worn continuously except for brief periods of removal for purposes of cleaning, replacement of the lens fluid reservoir, and examination and photography of the cornea. RESULTS: Five of the seven persistent corneal epithelial defects associated with Stevens-Johnson syndrome healed. The persistent corneal epithelial defects of four of these eyes re-epithelialized within 7 days, and a fifth healed in 27 days of gas-permeable scleral lens extended wear. A sixth persistent corneal epithelial defect that failed to heal initially re-epithelialized after a subsequent penetrating keratoplasty and gas permeable scleral lens extended wear. The seventh eye healed after 3 days of gas permeable scleral lens extended wear, but the persistent corneal epithelial defect subsequently recurred. Three of seven non-Stevens-Johnson syndrome persistent corneal epithelial defects re-epithelialized within 36 hours, 6 days, and 36 days, respectively. Of the six (six of 14) persistent corneal epithelial defects that failed to heal with a gas-permeable scleral lens extended wear, one subsequently healed after multiple amniotic membrane grafts. Microbial keratitis occurred in four eyes (four of 14) and graft failure in one eye, all of which required repeat penetrating keratoplasty. CONCLUSION: Extended wear of an appropriately designed gas-permeable scleral contact lens was effective in promoting the healing of persistent corneal epithelial defects in some eyes that failed to heal after other therapeutic measures. Re-epithelialization appears to be aided by a combination of oxygenation, moisture, and protection of the fragile epithelium afforded by the scleral lens. However, microbial keratitis represents a significant risk. PMID- 11004258 TI - Outcome of probing for congenital nasolacrimal duct obstruction in older children. AB - PURPOSE: To evaluate the role of probing in congenital nasolacrimal duct obstruction in children age 2 years and older and to establish factors predictive of the outcome. METHODS: The study was a single-center, prospective, interventional case series. Sixty patients with congenital nasolacrimal duct obstruction aged 24 months or older (range, 24 to 186 months; median, 33 months) presenting consecutively to the authors' institutional referral practice were studied. Probing of the nasolacrimal system under general anesthesia was the surgical intervention. Success of probing was the main outcome measure. Success was predefined as complete resolution of symptoms and signs (tearing, crusting, discharge, regurgitation on pressure over the lacrimal sac) of congenital nasolacrimal duct obstruction within 3 weeks of the procedure and continued remission at 6 months. Two attempts at probing were necessary before the procedure was declared a failure. RESULTS: One attempt at probing resulted in resolution in 73.3% (44 of 60) patients. Sixteen patients needed a repeat procedure. The overall success rate was 80% (48 of 60). Two specific types of obstructions of the nasolacrimal duct were recognized on probing: membranous and firm. Factors predictive of failure of probing were age older than 36 months (P <.0001); bilateral affection (P =.012); failed conservative therapy (P =.015); failed earlier probing (P <.0001); dilated lacrimal sac (P <.0001); and firm obstruction (P <.0001). CONCLUSION: Results indicate that probing is a viable primary surgical option for congenital nasolacrimal duct obstruction in children who present between 2 and 3 years of age, and identify factors predictive of poor prognosis. PMID- 11004259 TI - Immune-recovery uveitis in patients with cytomegalovirus retinitis taking highly active antiretroviral therapy. AB - PURPOSE: To investigate the clinical features associated with immune recovery in human immunodeficiency virus (HIV)-infected patients with cytomegalovirus retinitis who are taking highly active antiretroviral therapy. METHODS: Sixteen patients were evaluated prospectively at the National Eye Institute, Bethesda, Maryland. Evaluation included a medical history and a complete ophthalmologic examination. The examination included best-corrected visual acuity score measured by means of logarithmic charts, slit-lamp biomicroscopy, dilated retinal examination, retinal photography, and fluorescein angiography. Immune-recovery uveitis was defined as the ocular inflammation associated with clinical immune recovery in patients taking potent antiretroviral regimens. The ophthalmic characteristics of immune-recovery uveitis were identified, and their effect on visual acuity was statistically analyzed. RESULTS: The mean CD4+ T-lymphocyte count for the 16 patients taking highly active antiretroviral therapy at the time of evaluation was 393 cells/microl (range, 97-1,338 cells/microl). Immune recovery uveitis was characterized by vitreitis and optic disk and macular edema. Clinically important complications of immune-recovery uveitis included cataract and epiretinal membrane formation. The visual acuity scores were significantly worse in the 23 eyes with cytomegalovirus retinitis (mean, 67.2 letters, 20/50) than in the nine eyes without cytomegalovirus retinitis (mean, 89.8 letters, 20/16) (P <.001). Regression analysis showed that a lower visual acuity score was associated with the presence of moderate to severe macular edema on fluorescein angiography and vitreous haze (P < or =. 001). CONCLUSIONS: Immune-recovery uveitis is an important cause of visual morbidity in HIV-infected patients with cytomegalovirus retinitis in the era of highly active antiretroviral therapy. Although immune recovery associated with highly active antiretroviral therapy has allowed some patients to discontinue specific anticytomegalovirus therapy, the rejuvenated immune response can be associated with sight-threatening inflammation. PMID- 11004260 TI - Long-term posterior and anterior segment complications of immune recovery uveitis associated with cytomegalovirus retinitis. AB - PURPOSE: To identify and describe long-term posterior and anterior segment complications of immune recovery uveitis in patients with inactive cytomegalovirus retinitis who are undergoing highly active antiretroviral therapy mediated recovery of immune function. METHODS: A prospective cohort study at a university medical center. Twenty-nine eyes of 21 patients with immune recovery uveitis and inactive cytomegalovirus retinitis were followed for 14.5 to 116 weeks (median, 43 weeks) after diagnosis of immune recovery uveitis. RESULTS: Nine eyes of nine patients developed visually important complications involving the posterior segment, anterior segment, or a combination of both. Posterior segment complications included severe proliferative vitreoretinopathy in three eyes and spontaneous vitreous hemorrhage from avulsion of a blood vessel secondary to contraction of the inflamed vitreous in one eye. Proliferative vitreoretinopathy recurred in all cases after surgery, severely compromising the visual outcome. Anterior segment complications included posterior subcapsular cataracts with vision decrease in five eyes and persistent anterior chamber inflammation after cataract extraction, resulting in posterior synechiae and large visually important lens deposits in three eyes. CONCLUSION: Persistent inflammation in immune recovery uveitis may lead to vision-threatening complications, such as proliferative vitreoretinopathy, posterior subcapsular cataracts, and severe postoperative inflammation. Immune recovery uveitis is a chronic inflammatory syndrome that may result in complications months to years after the onset of inflammation. PMID- 11004261 TI - Three-dimensional observations of developing macular holes. AB - PURPOSE: To describe the morphologic features of idiopathic macular holes and vitreous traction during macular hole evolution. METHODS: We prospectively examined 89 eyes of 82 patients with idiopathic macular holes (stage 1, 15 eyes; stage 2, 16 eyes; stage 3, 50 eyes; stage 4, eight eyes) using optical coherence tomography. In addition to optical coherence tomography, scanning laser ophthalmoscopy was performed in all 15 eyes with stage 1 hole, six of 16 eyes with stage 2, and 19 of 50 eyes with stage 3. RESULTS: In stage 1 eyes, optical coherence tomography revealed retinal split or cystic changes at the fovea in 11 of 15 eyes (73%) and foveal retinal detachment in four eyes (27%). Two eyes with foveal cysts progressed to stage 2, and one developed a stage 3 hole. In one eye with retinal detachment, the detached retina thinned and developed dehiscence. Optical coherence tomography showed a vitreous cortex that was detached in the perifoveal area but attached on the fovea in 11 of the 15 stage 1 eyes. In stage 2 macular holes, retinal tissue extending from the perifoveal retina formed a flap. Scanning laser ophthalmoscopy demonstrated intraretinal radiating striae, which corresponded to a foveal cyst or perifoveal cystic changes. CONCLUSIONS: Macular holes start as retinal splits or foveal cysts in most cases. The anterior wall of the cyst serves as a flap in stage 2 and an operculum in stage 3 holes. Radiating striae correspond to retinal splits or cysts and presumably represent an elevation of Henle fiber. In a few macular holes, foveal detachment is the initial change. The detached retina thins and eventually develops a hole. In both courses, anterior traction of the slightly detached vitreous cortex appears to be a major contributing factor to macular hole formation. PMID- 11004262 TI - Scleral outfolding for macular translocation. AB - PURPOSE: To report experimental study and clinical observation of scleral outfolding for macular translocation. METHODS: In six human cadaver eyes, circumferential mattress sutures to create sclera infoldings were compared with radial-interrupted mattress sutures to create scleral outfoldings. In a 75-year old man with macular degeneration and choroidal neovascular membrane, radial interrupted mattress sutures were used for macular translocation. RESULTS: In the human cadaver eyes, circumferential mattress sutures for scleral infolding created an average decrease in corresponding internal anteroposterior retinal contour of 0.36 mm. Radial-interrupted mattress sutures for scleral outfoldings created an average decrease in the corresponding internal anteroposterior retinal contour of 4. 61 mm. The statistical significance of the difference between infoldings of the sclera versus outfoldings of the sclera had a P value of.0001. CONCLUSIONS: Initial experimental and clinical study suggests that radial interrupted mattress sutures may generate more shortening of the internal scleral surface and greater macular translocation than circumferential mattress sutures. Additional studies are needed to evaluate the long-term effects of radial interrupted mattress sutures and macular translocation for treatment of subfoveal choroidal neovascularization secondary to age-related macular degeneration. PMID- 11004263 TI - Erbium:YAG laser vitrectomy: clinical results. AB - PURPOSE: To evaluate the intraoperative use of a new erbium:yttrium aluminum garnet (YAG) laser vitrectomy system. METHODS: An erbium:YAG laser combined with an infusion-suction system (Wavelight Laser Technology, Erlangen, Germany), equipped with a flexible fiber optic and a hand piece with a 20-gauge end tip and a side opening of 0.6 mm was used. Cutting rates were 2 to 30 Hz, and energies were 20 to 40 mJ. Between January 1998 and January 1999 the erbium:YAG laser system was used in 67 consecutive patients (68 eyes) where vitrectomy was indicated. The patients had a complete eye examination before surgery and postoperatively at 1 to 3 days, 1 week, 3 weeks, and 6 weeks. During surgery, the total time needed for vitrectomy, the laser time itself, amount of energy used, cutting rate, suction, and perfusion, as well as manipulative difficulties and/or complications, were documented. RESULTS: The average laser time was 4.5 minutes for a basic vitrectomy with a setting of 20 mJ, 20 Hz, and suction 100 to 200 mm Hg. Hemorrhages, loose intravitreal, and preretinal membranes of different densities were cut well with adjustment of energy. After a minimal observation time of 6 months (median, 12.7 months) no laser-associated complications were found. CONCLUSIONS: The erbium:YAG laser, combined with an infusion-suction system, offers a new technology for vitreous surgery. Its advantages, compared with mechanical vitrectomy cutters, are higher cutting rates and the modulation of energy. Modifications of the end tip are needed to develop this system further. PMID- 11004264 TI - Elevated vitreous nitric oxide levels in patients with proliferative diabetic retinopathy. AB - PURPOSE: The aim of this study was to determine nitric oxide levels in the vitreous of patients with proliferative diabetic retinopathy. METHODS: Using the spectrophotometric method based on Griess reaction, we measured levels of nitrite, the stable product of nitric oxide, in the vitreous of 21 eyes of 21 patients who underwent vitrectomy for the treatment of proliferative diabetic retinopathy with tractional retinal detachment, prospectively. Three samples were excluded from the study because of blood contamination. The control group was made up of vitreous samples from 15 eyes of 15 normal cadavers and five eyes of five patients who were undergoing vitrectomy for macular hole surgery. RESULTS: Nitrite levels were 0. 524 +/- 0.27 microM and 0.383 +/- 0.17 microM in the vitreous of patients with proliferative diabetic retinopathy of diabetes type I and type II, respectively. In 15 cadaver eyes and five vitreous samples from patients who underwent macular hole surgery, nitrite levels were below the detection limit (less than 0.08 microM). There was no significant difference between nitrite levels in patients with type I and type II diabetes (P =.56), whereas there was a significant difference between diabetes groups and controls (P <. 00001). CONCLUSION: Vitreous nitric oxide levels are elevated in patients with proliferative diabetic retinopathy with tractional retinal detachment. Nitric oxide may play a role in the pathogenesis of proliferative diabetic retinopathy. PMID- 11004265 TI - Appearance of the optic disk and retinal nerve fiber layer in atherosclerosis and arterial hypertension: an experimental study in rhesus monkeys. AB - PURPOSE: To evaluate changes in the appearance of the optic nerve head and retinal nerve fiber layer of rhesus monkeys with chronic arterial hypertension and atherosclerosis. METHODS: Color stereoscopic fundus photographs of 25 eyes of 25 rhesus monkeys (mean age +/- SD of 20.4 +/- 1.87 years) with chronic experimental systemic arterial hypertension and atherosclerosis (for a mean duration of 89.1 +/- 39.1 months and 104.6 +/- 62.2 months, respectively) were morphometrically evaluated. They were compared with color stereoscopic fundus photographs of 17 eyes of 17 normal monkeys (mean age +/- SD of 19.76 +/- 2.19 years) without any detectable systemic or ocular disease. There was no significant difference in age between the two study groups (P =.22). RESULTS: In the atherosclerotic-arterial hypertensive group, visibility of the retinal nerve fiber layer was significantly (35.691 +/- 5.95 units vs 28.72 +/- 9.18 units, P =.009) less and frequency of localized retinal nerve fiber layer defects was significantly (six of 25 or 24% vs zero of 17 or 0%, P =.01) more than in the normal control group. The two groups did not differ significantly in size of the neuroretinal rim (P =.66), shape of the neuroretinal rim (P >.15), size of alpha (P >.44) and beta (P >.65) zones of parapapillary chorioretinal atrophy, or regional distribution of alpha and beta zones (P >.40). CONCLUSIONS: Chronic experimental arterial hypertension and atherosclerosis do not markedly change the size and shape of the neuroretinal rim or parapapillary atrophy; however, they do lead to reduced visibility of the retinal nerve fiber layer, with localized retinal nerve fiber layer defects indicating optic nerve damage. Thus, unlike glaucomatous optic neuropathy, experimental arterial hypertension and atherosclerosis are not associated with a significant change in the parapapillary atrophy or the neuroretinal rim of the optic disk despite the loss of nerve fibers. PMID- 11004266 TI - Plaque radiotherapy for management of ciliary body and choroidal melanoma with extraocular extension. AB - PURPOSE: To report the results of plaque radiotherapy of ciliary body and choroidal melanoma with extraocular extension. METHODS: This retrospective study of 17 eyes of 17 patients with ciliary body and choroidal melanoma demonstrated extraocular extension that was clinically evident but less than 3 mm in thickness. In 14 eyes, the extraocular extension was located anteriorly and in three posteriorly. The outcome of plaque radiotherapy in terms of intraocular and extraocular tumor responses and the development of metastasis were analyzed. RESULTS: At a median follow-up of 63 months (range, 23 to 164 months) after plaque radiotherapy, all patients showed control of both the intraocular and extraocular components of ciliary body and choroidal melanoma. In no case did the intraocular or extraocular portion of the tumor relapse. The initial median thickness of the intraocular component was 4.9 mm, and the final median thickness was 2.4 mm. The initial median thickness of the extraocular component was 2.0 mm, and the final median thickness was 1.0 mm. The initial median basal diameter of the extraocular component was 3.5 mm, and the final median basal diameter was 2.0 mm. Of 17 patients, 11 (10 with anterior and one with posterior extraocular extension) showed partial regression in base and thickness of the extraocular extension, and in six there was little or no change. In those with anterior extrascleral extension, the sclera remained intact without clinically observable necrosis, and the scleral fibers were gradually more visible as the tumor regressed. Three patients (18%) died of metastatic melanoma at a median interval of 38 months after plaque radiotherapy. CONCLUSIONS: Plaque radiotherapy appears to be a reasonable treatment option for selected cases of ciliary body and choroidal melanoma with clinically visible extraocular extension less than 3 mm in thickness. PMID- 11004267 TI - Immune recovery uveitis. PMID- 11004268 TI - Axenfeld-Rieger syndrome in the age of molecular genetics. AB - PURPOSE: To review the molecular genetics of Axenfeld-Rieger syndrome and related phenotypes and to discuss how this information might affect the way that we classify these disorders. METHODS: A review of historical and recent literature on Axenfeld-Rieger syndrome and related disorders. The review includes clinical and molecular genetic literature relevant to these phenotypes. RESULTS: Three chromosomal loci have recently been demonstrated to link to Axenfeld-Rieger syndrome and related phenotypes. These loci are on chromosomes 4q25, 6p25, and 13q14. The genes at chromosomes 4q25 and 6p25 have been identified as PITX2 and FKHL7, respectively. Mutations in these genes can cause a wide variety of phenotypes that share features with Axenfeld-Rieger syndrome. Axenfeld anomaly, Rieger anomaly, Rieger syndrome, iridogoniodysgenesis anomaly, iridogoniodysgenesis syndrome, iris hypoplasia, and familial glaucoma iridogoniodysplasia all have sufficient genotypic and phenotypic overlap that they should be considered one condition. CONCLUSIONS: Axenfeld-Rieger syndrome is a term that can be used to describe a variety of overlapping phenotypes. To date, at least three known genetic loci can cause these disorders. The single most important feature of these phenotypes is that they confer a 50% or greater risk of developing glaucoma. Currently there is a fairly arbitrary grouping of disorders into small categories. Considering all of these phenotypes under the heading of Axenfeld-Rieger syndrome will allow easier communication between clinicians and scientists and eliminate arbitrary and confusing subclassification. PMID- 11004270 TI - Surgical debulking of eyelid and anterior orbital plexiform neurofibromas by means of the carbon dioxide laser. AB - PURPOSE: To present the carbon dioxide laser as an effective tool for surgical debulking of eyelid and orbital neurofibromas. METHOD: Two patients with neurofibromatosis underwent surgical debulking of their eyelid and orbital plexiform neurofibromas by means of the carbon dioxide laser. RESULTS: Acceptable cosmetic results were obtained with the removal of eyelid and orbital neurofibromas with improved hemostasis and minimal destruction of surrounding tissue when compared with conventional methods of removal. CONCLUSIONS: The carbon dioxide laser may allow significant improvement in the removal of plexiform neurofibromas. PMID- 11004269 TI - Blepharoptosis surgery complicated by late suture migration. AB - PURPOSE: To report late suture migration as a complication of blepharoptosis surgery. METHOD: Case reports. RESULTS: After upper eyelid blepharoptosis repair, two eyes of two patients developed unusual foreign body reactions and ulceration in the conjunctiva caused by migration of nonabsorbable suture from the levator aponeurosis to the upper conjunctival fornix. The foreign body caused eyelid edema, papillary changes in the upper tarsal conjunctiva, recurrent blepharoptosis, and a foreign body sensation that started more than 3 months after the surgery and persisted until the suture was removed. The sutures, hidden in edematous conjunctiva of the superior fornix, eluded detection. CONCLUSIONS: Foreign body reaction secondary to suture migration is an uncommon complication of blepharoptosis repair. PMID- 11004271 TI - Q118X mutation of M1S1 gene caused gelatinous drop-like corneal dystrophy: the P501T of BIGH3 gene found in a family with gelatinous drop-like corneal dystrophy. AB - PURPOSE: To analyze BIGH3 and M1S1 genes in two Japanese brothers with gelatinous drop-like corneal dystrophy and five unaffected family members. METHODS: DNA was extracted, and each part of the two genes was amplified and directly sequenced. RESULTS: On the BIGH3 gene, a heterozygous P501T mutation was found in the elder brother and three unaffected family members. On the M1S1 gene, both brothers with gelatinous drop-like corneal dystrophy showed a homozygous Q118X mutation, whereas all unaffected members were heterozygous. CONCLUSIONS: The Q118X mutation of M1S1 gene caused gelatinous drop-like corneal dystrophy. Although the P501T of the BIGH3 gene found in this pedigree was precisely the one reported for lattice corneal dystrophy IIIA, no clinical feature was shown, even in the 85-year-old father. This fact shows that the P501T mutation for LCDIIIA has low penetrance. PMID- 11004272 TI - Marginal keratitis: an uncommon form of topical dorzolamide allergy. AB - PURPOSE: To report a case of marginal keratitis resulting from topical dorzolamide hypersensitivity. METHOD: Case report. RESULTS: A 68-year-old woman presented with bilateral marginal keratitis 2 weeks after commencing bilateral topical dorzolamide. One week after discontinuation of topical dorzolamide, the patient was asymptomatic with complete resolution of corneal infiltrates. CONCLUSIONS: Topical dorzolamide may cause a hypersensitivity reaction in the form of marginal keratitis. Discontinuation of the offending medication should result in complete resolution. PMID- 11004273 TI - Surgical management of recurrent iris stromal cyst. AB - PURPOSE: To report the clinical history and surgical management of recurrent congenital iris stromal cyst. METHOD: Case report. Argon laser endophotocoagulation and cryotherapy were used to treat a recurrent iris stromal cyst in an 18-month-old boy. RESULTS: At 43 months of follow-up, there was no recurrence or evidence of epithelial ingrowth. CONCLUSION: Treatment with argon laser endophotocoagulation and cryotherapy resulted in complete resolution of a recurrent congenital iris cyst. PMID- 11004274 TI - Acute angle-closure glaucoma after albuterol nebulizer treatment. AB - PURPOSE: To report acute angle-closure glaucoma associated with albuterol nebulizer treatment. METHODS: Case report and review of the relevant literature. RESULTS: In a 75-year-old woman with asthma, acute angle-closure glaucoma in the right eye was probably caused by local absorption of albuterol after nebulizer administration. CONCLUSION: As albuterol is widely used in the management of patients with asthma, increased awareness by health care professionals of the potential for acute angle-closure glaucoma secondary to albuterol may decrease the incidence of this complication. PMID- 11004275 TI - Ultrasound biomicroscopy in the diagnosis and management of pars planitis caused by caterpillar hairs. AB - PURPOSE: To report the use of ultrasound biomicroscopy in the detection of caterpillar hairs in the pars plana in a patient with unilateral pars planitis. METHOD: Ultrasound biomicroscopic imaging of the anterior segment of the eye. RESULTS: Ultrasound biomicroscopy located a hair in the posterior chamber at the first visit and five more in the pars plana 1 month later. This finding was confirmed intraoperatively. CONCLUSION: Ultrasound biomicroscopy is useful in the diagnosis and management of unilateral pars planitis of uncertain cause. PMID- 11004276 TI - Ocular side-effects from systemic HPMPC (Cidofovir) for a non-ocular cytomegalovirus infection. AB - PURPOSE: To report a case of anterior uveitis and ocular hypotony during HPMPC treatment of a cytomegalovirus infection without ophthalmic involvement. METHODS: A 61-year-old bisexual white man with AIDS presented with moderate anterior uveitis and ocular hypotony after 8 weeks of intravenous HPMPC and oral probenecid co-treatment of a presumed recurrence of cytomegalovirus encephalitis. RESULTS: There was no evidence of an ocular cytomegalovirus infection. Cidofovir related uveitis was diagnosed, and the HPMPC-probenecid co-treatment was switched to ganciclovir monotherapy. The ocular inflammation was successfully managed with topical steroids and mydriatics. CONCLUSIONS: This description of ophthalmic side effects from systemic HPMPC in the absence of ocular cytomegalovirus infection supports the hypothesis that anterior uveitis and ocular hypotony result from a direct interaction between the drug and the ciliary body. PMID- 11004277 TI - Labial adenocarcinoma after treatment with cyclosporin a in a patient with panuveitis. AB - PURPOSE: To report a case of labial basal cell adenocarcinoma in a patient with uveitis on treatment with cyclosporin A. METHOD: Case report. A 73-year-old woman with panuveitis and retinal vasculitis presented with a lump on her lip after 52 months of treatment with cyclosporin A. RESULT: Excision biopsy showed a labial adenocarcinoma. CONCLUSION: Malignancy can occur after long-term cyclosporin A treatment for uveitis. PMID- 11004278 TI - Acute megakaryoblastic leukemia in Down syndrome: orbital infiltration. AB - PURPOSE: To describe an uncommon ocular presentation of acute megakaryoblastic leukemia in a child with Down syndrome. METHOD: Case report. Initial manifestation of disease was bilateral proptosis with secondary exposure keratitis caused by leukemic infiltration of the orbits. RESULTS: Bone marrow biopsy and immunophenotyping established the diagnosis of acute megakaryoblastic leukemia (FAB-M7). The leukemia was treated successfully with chemotherapy, with resolution of proptosis. The patient remained in remission more than 1 year after cessation of treatment. CONCLUSIONS: Bilateral proptosis can be a presenting sign of acute megakaryoblastic leukemia, a malignancy associated with Down syndrome. PMID- 11004279 TI - Dot-and-fleck retinopathy in Alport syndrome caused by a novel mutation in the COL4A5 gene. AB - PURPOSE: To describe an unusual form of dot-and-fleck retinopathy in a slower progressive form of X-linked Alport syndrome, caused by a novel missense mutation in the COL4A5 gene. METHOD: Ophthalmic examination, polymerase chain reaction, and single-strand conformational polymorphism analysis of genomic DNA were performed in the proband. RESULTS: Ophthalmoscopy revealed classic dot-and-fleck retinopathy but located in an unusual site. A novel COL4A5 gene mutation changing glycine to cysteine at 177 was identified. CONCLUSIONS: Although there is no correlation between mutation site and the resulting phenotype in Alport syndrome, our findings suggest that further novel mutations and different ocular manifestations may be associated with Alport syndrome. PMID- 11004280 TI - Optical coherence tomography patterns of choroidal osteoma. AB - PURPOSE: To determine the optical coherence tomographic images that are commonly observed in eyes with choroidal osteoma (choroidal ossification). METHODS: Three patients with choroidal osteoma were examined by optical coherence tomography. RESULTS: We found two optical coherence tomographic patterns in the eyes with choroidal osteoma. First, multiple tracks of high refractivity were present posterior to the tumor lesion. Second, thick and irregular plate-like, high signal intensity areas were present in the choroid in the region of the tumor. CONCLUSIONS: Optical coherence tomography can be useful in the diagnosis of choroidal osteoma. PMID- 11004282 TI - Abstracts PMID- 11004281 TI - Amniotic membrane transplantation in acute chemical and thermal injury. AB - PURPOSE: To present a case of chemical injury and a case of thermal injury treated by amniotic membrane transplantation in acute phase. METHODS: Case reports. An eye with sodium hydroxide injury, opaque cornea, and limbal ischemia of more than 180 degrees and an eye with hot tea injury, opaque cornea, stromal edema, and scarring were treated by amniotic membrane transplantation within the first few weeks of injury. RESULTS: In the eye with sodium hydroxide injury, 4 months after amniotic membrane transplantation, the ocular surface is stable, superficial corneal scarring with vascularization is present, and visual acuity is 20/25. In the eye with thermal injury, 6 months after amniotic membrane transplantation, the ocular surface is stable, but there is superficial scarring and vascularization, and visual acuity is 20/20. CONCLUSIONS: Amniotic membrane transplantation can be considered in chemical injury with severe limbal ischemia and in severe thermal injury in acute phase. Long-term studies are warranted to evaluate further the efficacy of amniotic membrane transplantation in these clinical situations. PMID- 11004284 TI - Abstracts PMID- 11004283 TI - Abstracts PMID- 11004286 TI - Abstracts PMID- 11004285 TI - Abstracts PMID- 11004287 TI - Long-term intraocular pressure-lowering efficacy and safety of timolol maleate gel-forming solution 0.5% compared with Timoptic XE 0.5% in a 12-month study. AB - PURPOSE: To evaluate the long-term intraocular pressure-lowering efficacy and safety of timolol maleate gel-forming solution 0.5% (Timolol GFS 0.5%, Alcon Research Ltd, Fort Worth, Texas) compared with Timoptic XE 0.5% (Merck, Inc, West Point, Pennsylvania) in patients with open-angle glaucoma or ocular hypertension. METHODS: Two hundred forty-one patients with open-angle glaucoma or ocular hypertension, who had intraocular pressure between 22 and 36 mm Hg in at least one eye, were randomly assigned in a 2:1 ratio to receive either Timolol GFS 0.5% once daily or Timoptic XE 0.5% once daily, in a 12-month randomized, multicenter, double-masked, prospective study. The primary efficacy variable was mean trough intraocular pressure measured at 8:00 AM, approximately 24 hours after dosing. RESULTS: The Timolol GFS 0.5% group produced significant trough intraocular pressure reductions from a baseline of 4.5 to 5.2 mm Hg (P =.0001), compared with reductions of 4.1 to 5. 3 mm Hg (P =.0001) in the Timoptic XE 0.5% group. The difference in mean intraocular pressure between the two treatments was 0.9 mm Hg or less, and the upper 95% confidence limit between groups was 0.92 mm Hg or less at all time points, demonstrating both clinical and statistical equivalence. A similar percentage of patients in the Timolol GFS 0.5% group (71%) and Timoptic XE group (72%) had clinically relevant reductions in intraocular pressure. There was no significant difference in the safety profiles of the two treatments. CONCLUSION: Both treatments were clinically effective in lowering intraocular pressure and maintaining the reductions over long-term use. Timolol GFS 0.5% is a safe and effective therapy for open-angle glaucoma or ocular hypertension and is both clinically and statistically equivalent to Timoptic XE 0.5% in reducing intraocular pressure. PMID- 11004288 TI - Effects of latanoprost and timolol-XE on hydrodynamics in the normal eye. AB - PURPOSE: To compare the effects of latanoprost and timolol-XE on ocular pressure and perfusion in healthy adults, with respect to episcleral venous pressure. METHODS: A double-masked, placebo-controlled crossover study of weeklong bedtime treatment with one drop of drug, with placebo contralaterally, followed by a 3 week washout and alternate-drug/contralateral-placebo repeat. Intraocular pressure was measured by applanation and by pneumotonometry, providing pulsatile ocular circulatory estimates. Measurements of episcleral venous pressure were obtained (Friberg method). RESULTS: Twenty subjects participated (five men, 15 women; mean age, 39 years (range, 21 to 55 years); mean baseline intraocular pressure, 13.4 mm Hg). A greater decrease in intraocular pressure was seen among these subjects the morning after initiating treatment with latanoprost (-2.0 mm Hg; P <.0001) than with timolol-XE (-0.9 mm Hg; P =.051) (latanoprost versus timolol DeltaP =.008). This ocular hypotensive effect remained significant that evening with latanoprost (-3.2 mm Hg; P <.0001) but not with timolol XE (-1.0 mm Hg; P =.2). By the morning of day 8, mean intraocular pressure remained 3.2 mm Hg below baseline with latanoprost and 2.3 mm Hg below baseline with timolol-XE (P <.0001 for both drugs). Neither drug altered episcleral venous pressure. Among a subgroup of nine subjects with comparable intraocular pressure reductions with the two drugs, latanoprost treatment was associated with a 16.7% increase in mean pulsatile ocular blood flow (P =.04) through the weeklong treatment interval, consistently higher than during timolol-XE treatment of the same subjects. CONCLUSION: Latanoprost caused an overnight decrease in intraocular pressure in normotensive normal eyes, and both drugs significantly reduced intraocular pressure within 1 week. Intraocular pressure remained higher than episcleral venous pressure throughout treatment with both drugs. Latanoprost was associated with enhanced pulsatile ocular perfusion not seen with timolol-XE treatment. PMID- 11004289 TI - Learning effect, short-term fluctuation, and long-term fluctuation in frequency doubling technique. AB - PURPOSE: To evaluate short-term and long-term fluctuations and learning effects in healthy subjects tested with the frequency doubling technique, which is based on a low spatial frequency and a high temporal frequency stimulus. METHODS: Twenty-three healthy adult subjects were consecutively recruited from volunteers, and 20 subjects finished the study. All the visual fields were assessed by the frequency doubling technique, program C-20, full threshold. The frequency doubling technique presents stimuli on a black-and-white video monitor with specialized control circuitry interfaced to a microprocessor. During program C 20, full threshold, 17 points are tested, one round point centrally and 16 square ones in the periphery up to 20 degrees of eccentricity. Each stimulus consists of a 0.25-cycle/degree sinusoidal grating undergoing a 25-Hz counterphase flicker. One eye of each patient was chosen at random. Each subject was tested once in the first session, three times in the second session, and once in the third and fourth sessions. Both short-term and long-term fluctuation were studied either as the average fluctuation value of all the points tested or as a point-to-point fluctuation. To study the learning effect, the results of the first session were compared with those of the second, the third, and the fourth sessions. RESULTS: The average mean sensitivity of the three examinations of the second session was 30.4 +/- 1.24 dB, and the average short-term fluctuation of all the subjects was 2.16 +/- 0.5 dB. The short-term fluctuation of each point tested ranged from 1.4 to 3.4 dB. The average mean sensitivity for all the sessions was 32.4 +/- 1.14 dB, with an average long-term fluctuation of 3.23 +/- 0.5 dB, and the long-term fluctuation of each tested point ranged from 2.5 to 4.4 dB. A mild learning effect was found between the first section and the others. CONCLUSION: Short-term and long-term fluctuations were similar to those known to occur with the conventional threshold perimetry when they were compared with the literature data. A learning effect was also observed and should be taken into account for the clinical use of this test. PMID- 11004290 TI - Age-dependent prevalence of mutations at the GLC1A locus in primary open-angle glaucoma. AB - PURPOSE: To screen a population with primary open-angle glaucoma for mutations in the gene that encodes the trabecular meshwork inducible glucocorticoid response protein (TIGR), also known as myocilin (MYOC). METHODS: Ophthalmologic information was collected for study subjects with primary open-angle glaucoma and their relatives. Mutation screening of 74 primary open-angle glaucoma probands was conducted by sequencing TIGR/MYOC coding sequence and splice sites. RESULTS: In 23 families we detected 13 nonsynonymous sequence changes, nine of which appear to be mutations likely to cause or contribute to primary open-angle glaucoma. Two mutations, Arg272Gly and Ile499Ser, and one nonsynonymous sequence variant, Asn57Asp, are novel. We found mutations in nine of 25 juvenile glaucoma probands (36%) and two of 49 adult-onset glaucoma probands (4%). Age classification of families rather than individual probands revealed mutations in three of nine families with strictly juvenile primary open-angle glaucoma (33%), and no mutations in 39 families with strictly adult-onset primary open-angle glaucoma (0%). In families with mixed-onset primary open-angle glaucoma containing both juvenile primary open-angle glaucoma and adult-onset primary open angle glaucoma cases, we found mutations in eight of 26 families (31%). CONCLUSIONS: Our data suggest that Gly252Arg, Arg272Gly, Glu323Lys, Gln368STOP, Pro370Leu, Thr377Met, Val426Phe, Ile477Asn, and Ile499Ser are likely to play roles that cause or contribute to the etiology of autosomal dominant primary open angle glaucoma. Our finding of more TIGR/MYOC mutations in families with mixed onset primary open-angle glaucoma than in the families with strictly adult-onset primary open-angle glaucoma implies that the presence of relatives with juvenile primary open-angle glaucoma in a family could be used as a basis for identifying a subset of the population with adult-onset primary open-angle glaucoma with higher prevalence of TIGR/MYOC mutations. To address this issue, and to refine estimations of mutation prevalence in these age-defined subpopulations, prospective study of a larger population ascertained entirely through adult-onset primary open-angle glaucoma probands will be needed. PMID- 11004291 TI - Vitrectomy for diffuse diabetic macular edema associated with a taut premacular posterior hyaloid. AB - PURPOSE: To evaluate the role of vitrectomy in eyes with diffuse diabetic macular edema associated with a taut posterior hyaloid. METHODS: Records of 55 eyes of 50 patients with diabetic retinopathy and diffuse clinically significant diabetic macular edema who underwent vitrectomy with stripping of the premacular posterior hyaloid were reviewed. In all 55 eyes, diffuse diabetic macular edema was present on contact lens examination and confirmed with fluorescein angiography. On fundus examination, the premacular posterior hyaloid was attached and appeared taut. RESULTS: The mean preoperative best-corrected visual acuity was 20/160, and the mean final best-corrected visual acuity was 20/80 (P <.0001, Wilcoxon signed rank test), with 27 (49.1%) of the 55 eyes demonstrating improvement in best-corrected visual acuity of 2 or more lines. Fifty-two (94.5%) of the 55 vitrectomized eyes showed improvement in clinically significant macular edema and in 45 eyes (81.8%) the macular edema resolved completely during a mean period of 4.5 months (range, 1 to 13 months). Eyes with macular ischemia and preoperative best-corrected visual acuity of 20/200 or less tended to respond less favorably to vitrectomy than eyes lacking these characteristics. All eyes had at least 6 months of follow up after surgery, with a mean follow-up of 23.2 months. CONCLUSION: In eyes with persistent diffuse diabetic macular edema with a taut premacular posterior hyaloid face unresponsive to laser therapy, vitrectomy with removal of the posterior hyaloid appears to be beneficial in some cases. Careful selection of eyes with favorable preoperative clinical characteristics may improve surgical outcomes. PMID- 11004292 TI - Submacular surgery to remove choroidal neovascularization associated with central serous chorioretinopathy. AB - PURPOSE: To report the results of submacular surgery for removal of choroidal neovascularization associated with central serous chorioretinopathy. METHODS: Ten eyes of nine consecutive patients with central serous chorioretinopathy and subfoveal or juxtafoveal choroidal neovascularization underwent pars plana vitrectomy with removal of the choroidal neovascular membrane between January 1994 and January 1999. RESULTS: All 10 eyes (nine patients) were followed postoperatively for at least 6 months. The mean postoperative follow-up was 23 months (range, 6 to 56.5 months). The patients were followed for an average of 6.2 months from the time of symptoms to the removal of the choroidal neovascularization. Preoperative mean best-corrected visual acuity was 20/100 (range, 20/25 to 20/400), and postoperative best-corrected mean visual acuity was 20/60 + 2 (range, 20/20 to 20/400). Seven eyes had improved postoperative visual acuity, with an average of 3.4 lines gained. Of the three eyes that had worse acuity, an average of 1.3 lines of visual acuity was lost; final mean postoperative visual acuity was 20/80 + 1 (range, 20/25 to 20/400). Of the six eyes with symptoms of less than 3 months' duration, four had a final visual acuity of 20/50 or better. All three eyes with 20/300 or worse final visual acuity had loss of foveal retinal pigment epithelium after surgery; the remaining eyes had preserved retinal pigment epithelium with a visual acuity of 20/70 or better. Two eyes had intraoperative peripheral retinal tears, and two eyes had recurrence of the choroidal neovascular complex. CONCLUSIONS: The anatomic and visual results in eyes with choroidal neovascularization associated with central serous chorioretinopathy are modestly encouraging and suggest that submacular surgery for choroidal neovascularization in patients with central serous chorioretinopathy is a treatment option that may salvage good macular function in some eyes. PMID- 11004293 TI - Tomographic features of early macular hole closure after vitreous surgery. AB - PURPOSE: To report the optical coherence tomographic features of macular hole closure in the first months after vitreous surgery. METHODS: We studied prospectively the tomographic features of 28 eyes (28 patients) with idiopathic macular holes before and after vitreous surgery by optical coherence tomography. We compared the best-corrected visual acuity levels with the postoperative tomographic features. RESULTS: The 25 eyes with successfully sealed macular holes had one of two tomographic features within 1 month postoperatively: simple closure (normal foveal configuration) in 14 eyes (56%) or a bridge formation at the fovea that mimicked a foveal retinal detachment in 11 eyes (44%). It took an average of 2.0 months (range, 0.8 to 3.5 months) for the bridge tissue to attach to the retinal pigment epithelium. Best-corrected visual acuity quickly improved in the former group; visual improvement began 1 month after attachment of the bridge tissue in the latter group. A closed hole reopened 4 months postoperatively in one eye with a bridge formation. CONCLUSIONS: Idiopathic macular holes have one of two patterns early after surgical closure, simple closure or a bridge formation. Visual improvement starts after the fovea assumes a normal configuration. The bridge formation appears to reflect an early phase and fragile condition in the anatomic closure of macular holes. PMID- 11004294 TI - A new locus for dominant drusen and macular degeneration maps to chromosome 6q14. AB - PURPOSE: To report the localization of a gene causing drusen and macular degeneration in a previously undescribed North American family. METHODS: Genetic mapping studies were performed using linkage analysis in a single family with drusen and atrophic macular degeneration. RESULTS: The clinical manifestations in this family ranged from fine macular drusen in asymptomatic middle-aged individuals to atrophic macular lesions in two children and two elderly patients. We mapped the gene to chromosome 6q14 between markers D6S2258 and D6S1644. CONCLUSIONS: In a family with autosomal dominant drusen and atrophic macular degeneration, the gene maps to a 3.2-cM region on chromosome 6q14. This locus appears to be distinct from, but adjacent to, the loci for cone-rod dystrophy 7 (CORD7) and North Carolina macular dystrophy (MCDR1). Future identification of the gene responsible for the disease in this family will provide a better understanding of macular degeneration. PMID- 11004295 TI - Clinical spectrum of chromosome 6-linked autosomal dominant drusen and macular degeneration. AB - PURPOSE: To describe the clinical phenotype and the intrafamilial variation in retinal findings in a North American family with an autosomal dominant drusen disorder that maps to chromosome 6q14. METHODS: Ophthalmic examinations were carried out on participating family members. Fundus photographs were obtained whenever possible. Electroretinography was performed on the proband and her father. Blood was drawn for DNA analysis. RESULTS: Twelve family members had drusen and/or atrophic macular degeneration. The disease in asymptomatic young adults is characterized by fine drusen that are most conspicuous in the macula. The proband presented at 3 years of age with atrophic maculopathy and drusen. Her cousin was found to have atrophic macular lesions and drusen in the first year of life. Two older affected individuals have reduced vision from cicatricial and atrophic macular changes. The gene for the disease was mapped to chromosome 6q14 and appears to be adjacent to but distinct from the locus for North Carolina macular dystrophy. CONCLUSIONS: There is extreme variability in the clinical expression of this dominant form of drusen and macular degeneration. Most young adults have fine macular drusen and good vision. Affected infants and children may have congenital atrophic maculopathy and drusen. There is historical evidence of progression of the disease in late adulthood with moderate visual loss. PMID- 11004296 TI - Lymphocytic choriomeningitis virus: an underdiagnosed cause of congenital chorioretinitis. AB - PURPOSE: To elucidate the role and clinical spectrum of congenital lymphocytic choriomeningitis virus infection as a cause of chorioretinopathy, congenital hydrocephalus, and macrocephaly or microcephaly in the United States. METHODS: We performed complete ophthalmologic surveys of all residents at Misericordia, a home for the severely mentally retarded in Chicago, and prospectively evaluated all patients with chorioretinitis or chorioretinal scars during a 36-month period at Children's Memorial Hospital, also located in Chicago. Sera for patients demonstrating chorioretinal scars (a sign of intrauterine infection) were tested for Toxoplasma gondii, rubella virus, cytomegalovirus, and herpes simplex virus and lymphocytic choriomeningitis virus antibodies. RESULTS: Four of 95 patients examined at the home had chorioretinal scars, and two of these patients had normal T. gondii, rubella virus, cytomegalovirus, and herpes simplex virus titers and dramatically elevated titers for lymphocytic choriomeningitis virus. Three of 14 cases of chorioretinitis at the hospital had normal T. gondii, rubella virus, cytomegalovirus, and herpes sim-plex virus titers and elevated lymphocytic choriomeningitis virus antibody titers. (A fourth case, diagnosed in 1996, was reported 2 years ago.) CONCLUSIONS: Lymphocytic choriomeningitis virus was responsible for visual loss in two of four children secondary to chorioretinitis in a population of severely retarded children. The six new cases of lymphocytic choriomeningitis virus chorioretinitis identified in these two populations over the last 3 years, compared with the total number ever reported in the United States (10 cases), suggests that lymphocytic choriomeningitis virus may be a more common cause of congenital chorioretinitis than previously believed. Because its consequences for visual and psychomotor development are devastating, we conclude that the workup for congenital chorioretinitis should include lymphocytic choriomeningitis virus serology, especially if T. gondii, rubella virus, cytomegalovirus, and herpes simplex virus titers are negative. PMID- 11004297 TI - Isolated comitant esotropia and Chiari I malformation. AB - PURPOSE: To report four patients with isolated comitant esotropia and Chiari I malformation and discuss the most appropriate management. METHODS: Case reports and literature review. RESULTS: All four patients (5, 14, 16, and 37 years of age) presented with an isolated comitant esotropia that led to the diagnosis of Chiari I malformation. The first two patients underwent uncomplicated neurosurgical decompression of their malformation, followed by complete resolution of their esotropia. The third patient underwent strabismus surgery and experienced initial resolution of the esotropia, but eventual recurrence resulted in the strabismus surgery being repeated 5 years later. The fourth patient had strabismus surgery with resolution of the esotropia but only 2 months of follow up. CONCLUSION: Although management of patients with Chiari I malformation and severe neurologic findings typically includes surgical decompression, management is less straightforward in cases with subtle findings or in which ocular findings are isolated. The decision to perform neurosurgical decompression or strabismus surgery should still be made on a case-by-case basis, with the understanding that strabismus surgery may provide only temporary ocular alignment. PMID- 11004298 TI - Monitoring patients taking oral carbonic anhydrase inhibitors. AB - Although no prospective studies have been done and no recommendations can be reasonably evidenced based, we believe that our "alternative views" are reason able, taking into account the serious nature of aplastic anemia and its more favorable prognosis when this disorder is treated early. Because aplastic anemia represents only about one half of possible CAl blood dyscrasias, early recognition of non-aplastic anemia dyscrasias alone Justifies hematologic screening. PMID- 11004299 TI - International Federation of Ophthalmological Societies statutes and regulations. AB - PURPOSE: To provide an understanding of the structure, role, and responsibility of the International Federation of Ophthalmological Societies, the International Council of Ophthalmology, and the International Congress of Ophthalmology. These established entities have recently codified their statutes and regulations and registered them in Zurich, Switzerland. METHODS: The International Council of Ophthalmology, which serves as the executive body of the International Federation of Ophthalmological Societies, used historical operating documents to prepare the statutes and regulations, which were reviewed and adopted by the International Federation of Ophthalmological Societies at the 28th International Congress of Ophthalmology in Amsterdam on June 23, 1998. RESULTS: The statutes and regulations of the International Federation of Ophthalmological Societies, the International Council of Ophthalmology, and the International Congress of Ophthalmology are available to supranational ophthalmological organizations, national ophthalmological societies, other ophthalmological organizations, and individual ophthalmologists. CONCLUSIONS: The statutes and regulations of the International Federation of Ophthalmological Societies, the International Council of Ophthalmology, and the International Congress of Ophthalmology provide a basis for an organizational structure in international ophthalmology. The International Congress of Ophthalmology, first held in 1857, is the longest continuing international meeting in medicine. The International Council of Ophthalmology was established in 1927, and the International Federation of Ophthalmological Societies was formed in 1933. These organizations coordinate the International Congress of Ophthalmology, which convenes with the International Federation of Ophthalmological Societies every 4 years. The International Council, as the executive body, meets annually. The International Council of Ophthalmology has for decades worked to coordinate and facilitate interchange, education, and standards in international ophthalmology. Currently, the International Council of Ophthalmology, working with other international organizations, is creating an international ophthalmology strategic plan, which includes focus on ophthalmic training, continuing education, advocacy for the preservation and restoration of vision, clinical guidelines, and research. PMID- 11004300 TI - Blepharoptosis-induced superior keratoconus. AB - PURPOSE: This clinical case report demonstrates blepharoptosis-induced corneal steepening and its subsequent resolution after blepharoptosis surgery. METHODS: A 62-year-old man complaining of blurred vision without apparent cause on clinical examination underwent keratometry and computerized corneal topography, which revealed superior corneal steepening in both eyes. Bilateral upper eyelid blepharoptosis surgery was performed. RESULTS: Three months after blepharoptosis surgery, repeat computerized corneal topography revealed normal corneal contours with improved vision in both eyes. CONCLUSIONS: Blepharoptosis is a common condition that may induce superior corneal ectasia that is not evident by manifest refraction, slit-lamp examination, or keratometry. Computerized corneal topography can help detect such subtle corneal abnormalities and guide therapy. PMID- 11004301 TI - Air bag-induced corneal flap folds after laser in situ keratomileusis. AB - PURPOSE: We describe a case of air bag-induced ocular trauma resulting in folds in the corneal flap 3 weeks after laser in situ keratomileusis. METHODS: Case report. Three weeks after laser in situ keratomileusis, a 20-year-old man was involved in a motor vehicle accident and sustained blunt trauma to the right eye, which caused corneal flap folds, corneal edema, anterior chamber cellular reaction, and Berlin retinal edema. RESULTS: Six weeks after laser in situ keratomileusis, persistent flap folds necessitated re-operation with lifting of the flap and repositioning. One week after the procedure, the visual acuity improved to 20/20-2, and the folds had cleared. CONCLUSION: Trauma after laser in situ keratomileusis may produce folds in the corneal flap. With persistence of these folds, management by lifting and repositioning the corneal flap may be necessary to permit recovery of visual acuity. PMID- 11004302 TI - Compound nevus of the cornea simulating a foreign body. AB - PURPOSE: To report a corneal nevus, without conjunctival involvement, that clinically simulated a metallic foreign body. METHODS: Case report with clinicopathologic correlation. RESULTS: A 22-year-old man developed a superficial pigmented lesion of his left cornea. It was suspected to be a chronically retained foreign body, but a melanocytic neoplasm could not be excluded. The lesion was removed by lamellar keratectomy and proved histopathologically to be a compound nevus of the cornea, without conjunctival involvement. CONCLUSIONS: Melanocytic nevus can rarely occur in the cornea. Involvement of the cornea by a melanocytic lesion does not necessarily imply a malignant melanoma. PMID- 11004303 TI - Iris color and cataract: the Blue Mountains Eye Study. AB - PURPOSE: To investigate the relationship between eye color and cataract. METHODS: A population-based cross-sectional study (N = 3654) was conducted near Sydney, Australia. Lens photographs were graded for cortical, nuclear, and posterior subcapsular cataract. Iris color was assessed at slit-lamp examination by comparison with four photographic standards. RESULTS: Eyes with dark brown irises were more likely to have nuclear (adjusted odds ratio, 1.59; 95% confidence interval [CI], 1.03 to 2.28) or posterior subcapsular cataract (adjusted odds ratio, 2.50; 95% CI, 1.57 to 3.98) than eyes with lighter-colored irises. CONCLUSIONS: People with dark brown eyes are at increased risk of cataract and should be encouraged to protect their eyes from direct exposure to sunlight. PMID- 11004304 TI - Tonic pupil associated with congenital neuroblastoma, Hirschsprung disease, and central hypoventilation syndrome. AB - PURPOSE: To report the association of tonic pupil, congenital neuroblastoma, Hirschsprung disease, and central hypoventilation syndrome. METHODS: Case reports. RESULTS: Two infants with central hypoventilation syndrome and Hirschsprung disease were evaluated for dilated, nonreactive pupils present since birth. In both cases, pharmacologic testing with dilute pilocarpine confirmed denervation supersensitivity consistent with tonic pupil. The diagnosis of congenital neuroblastoma was subsequently established in both infants. CONCLUSIONS: The close association of these conditions in these two young children supports the concept of a common neural crest abnormality being present. A careful systemic evaluation to rule out congenital neuroblastoma should be performed in any young child who presents with tonic pupil in the setting of central hypoventilation syndrome and Hirschsprung disease. PMID- 11004305 TI - Systemic toxicity of topical and periocular corticosteroid therapy in an 11-year old male with posterior uveitis. AB - PURPOSE: To report a case of systemic corticosteroid toxicity resulting from topical and periocular therapy. METHODS: Treatment and follow-up of an 11-year old male with uveitis are illustrated. Initial presentation of the patient was bilateral iridocyclitis, for which he was treated with prednisolone acetate 1% every 2 hours for 6 months. Subsequently, posterior uveitis developed, necessitating posterior subtenon injections. RESULTS: After initial topical corticosteroid therapy, the patient developed a cushingoid habitus accompanied by increased lanugo hair, acanthosis nigricans, posterior subcapsular lens opacities, and increased intraocular pressure. Cushingoid stigmata worsened after administration of posterior subtenon injection of corticosteroids. The patient's truncal obesity worsened, and his linear growth stopped. CONCLUSIONS: Systemic toxic effects may develop as a result of topical and local use of ophthalmic corticosteroid preparations in susceptible patients. PMID- 11004306 TI - Reversible bull's-eye maculopathy associated with intravitreal fomivirsen therapy for cytomegalovirus retinitis. AB - PURPOSE: To report two cases in which a bull's eye maculopathy developed after intravitreal injection of fomivirsen. METHODS: Case reports. RESULTS: A 50-year old man with acquired immunodeficiency syndrome (AIDS) and refractory cytomegalovirus retinitis developed bull's-eye pigmentary changes in the macula of the right eye after initiating therapy with fomivirsen (Vitravene; CIBA Vision, Atlanta, Georgia) intravitreal injections. These pigmentary changes resolved upon cessation of treatment. A 36-year-old man with AIDS and refractory bilateral cytomegalovirus retinitis developed bull's-eye pigmentary changes in both eyes during bilateral intravitreal treatment with fomivirsen. Vision was not affected. These changes resolved after treatment with fomivirsen was stopped. CONCLUSION: Fomivirsen, a new medication for the treatment of refractory cytomegalovirus retinitis, may cause a bull's-eye maculopathy in some patients. The bull's-eye maculopathy is reversible and does not appear to affect vision. PMID- 11004307 TI - Clonally accumulating T cells in the anterior chamber of Behcet disease. AB - PURPOSE: To clarify the nature of infiltrating T cells into the anterior chamber of a patient with Behcet disease. METHODS: Aqueous humor was obtained from a patient with ocular Behcet disease by paracentesis. RNA isolated from the cells in aqueous humor was reverse transcribed into complementary DNA. Complementary DNA encoding the variable (V) diversity (D) joining (J) (junctional) region of T cell receptor beta chain V domain (TCR BV) chain was amplified by T-cell receptor BV family polymerase chain reaction. RESULTS: Polymerase chain reaction Southern blot analysis showed that T-cell receptor BV3, BV5, and BV7 were dominantly expressed on ocular T cells of this patient. In addition, DNA sequencing revealed that monoclonal or oligoclonal T-cell accumulation was found in T-cell receptor BV3(+), BV5(+), and BV7(+) T cells. CONCLUSION: These findings suggest that some T cells infiltrating into the anterior chamber of a patient with ocular Behcet disease expand by antigen-driven stimulation, indicating the pivotal role of T cells in the pathogenesis of ocular Behcet disease. PMID- 11004309 TI - Choroidal neovascularization with granulomatous inflammation in ocular histoplasmosis syndrome. AB - PURPOSE: To report pathologic examination of an excised choroidal neovascular membrane in a patient with ocular histoplasmosis syndrome that demonstrated granulomatous inflammation. METHOD: Case report. A 50-year-old woman with sudden vision loss in her left eye demonstrated clinical and fluorescein angiographic findings characteristic of choroidal neovascularization secondary to ocular histoplasmosis syndrome. RESULTS: Histopathologic examination of the surgically excised choroidal neovascular membrane disclosed granulomatous inflammation. CONCLUSIONS: This case suggests an important role of mononuclear phagocytic cells as primary mediators of angiogenesis or modifiers of choroidal neovascularization. This association of choroidal neovascularization with granulomatous inflammation did not respond to treatment with systemic corticosteroids. PMID- 11004308 TI - Lymphocytic choriomeningitis virus chorioretinitis mimicking ocular toxoplasmosis in two otherwise normal children. AB - PURPOSE: To report unilateral macular lesions, mimicking toxoplasmic scars, in two children with serological evidence for lymphocytic choriomeningitis virus infection. METHODS: Case reports. RESULTS: Patients were 4 and 5 years old, with negative toxoplasma serologies and no sign of rubella, cytomegalovirus, or herpes simplex infection (TORCH evaluation). Lymphocytic choriomeningitis virus infection was detected in both cases by enzyme-linked immunosorbent assay and confirmed by Western immunoblotting. The modes of infection were unknown; no history of symptomatic systemic lymphocytic choriomeningitis virus infection was reported, and lymphocytic choriomeningitis virus serologies were negative in the mothers of the patients. Neurological examinations and brain magnetic resonance imaging were normal. CONCLUSION: Our observations suggest that chorioretinal scars can be an isolated manifestation of lymphocytic choriomeningitis virus infection. PMID- 11004310 TI - Chorioretinal involvement in primary systemic nonfamilial amyloidosis. AB - PURPOSE: To report a case of primary systemic nonfamilial amyloidosis studied by fluorescein angiography and indocyanine green angiography. METHODS: Case report. A 59-year-old woman with primary systemic nonfamilial amyloidosis presented bilateral diffuse deep hemorrhages and pigmentary mottling at the posterior pole. RESULTS: On fluorescein angiography bilateral diffuse areas of hypofluorescence were present. Indocyanine green angiography showed large hypofluorescent areas with hypofluorescent lines in the midperiphery and hyperfluorescent streaks in the peripapillary area. CONCLUSIONS: In this case of primary systemic nonfamilial amyloidosis, diffuse bilateral chorioretinal abnormalities included hemorrhages and pigmentary mottling at the posterior pole, with hypofluorescent areas on fluorescein angiography and indocyanine green angiography, as well as hypofluorescent lines in the midperiphery. PMID- 11004311 TI - Traumatic displacement of the globe into the ethmoid sinus. AB - PURPOSE: To describe displacement of the globe into the ethmoid sinus after an orbital trauma. METHOD: Case report. A 58-year-old man sustained trauma of the left eye and orbit, which resulted in displacement of the globe into the ethmoid sinus. One day after injury, surgery was performed to restore the intact globe into position within the orbit. RESULTS: After operation, the globe held in its anatomical site, and 10 months after surgery the visual acuity was 20/100, slight pallor of the optic disk was present, and there was no limitation of the horizontal and vertical ductions. CONCLUSIONS: To our knowledge, this is the first case of traumatic displacement of the globe into the ethmoid sinus with satisfactory restoration of normal globe position and preservation of vision. PMID- 11004312 TI - Bilateral papilledema from a massive intracranial epidermoid cyst. AB - PURPOSE: To report an unusual case of bilateral papilledema from a large intracranial epidermoid cyst. METHODS: Case report. RESULTS: A 26-year-old man presented with visual loss, bilateral papilledema, and only a few neurological symptoms. Magnetic resonance imaging disclosed such a large lesion that his right cerebral hemisphere was compressed to one half its normal size. Histopathologic examination of the completely removed tumor revealed an epidermoid cyst. CONCLUSION: This unique case involved a patient with bilateral papilledema caused by a huge intracranial epidermoid cyst. Epidermoid tumors should be considered in the differential diagnosis of papilledema. PMID- 11004314 TI - Hallucinations in Parkinson's disease. prevalence, phenomenology and risk factors. Fenelon G* mahieux F, huon R, Ziegler M. Brain 2000;123:733-745 PMID- 11004313 TI - Physicians and the pharmaceutical industry: is a gift ever just a gift? wazana A.* JAMA 2000;283:373-380 PMID- 11004316 TI - Recovery from optic neuritis is associated with a change in the distribution of cerebral response to visual stimulation: a functional magnetic resonance imaging study. Werring DJ, bullmore ET, toosy AT, miller DH, barker GJ, MacManus DG, brammer MJ, giampietro VP, brusa A, brex PA, moseley IF, plant GT, McDonald WI, thompson AJ. * J neurol neurosurg psychiatry 2000;68:441-449 PMID- 11004315 TI - Visual loss after spine surgery: a survey. Cheng MA, sigurdson W, tempelhoff R, lauryssen C.* neurosurgery 2000;46:625-631 PMID- 11004318 TI - Is CS gas dangerous? fraunfelder FT.* Br med J 2000; 320:458-459 PMID- 11004317 TI - Foveal relocation by redistribution of the neurosensory retina. Wong D* lois N. Br J ophthalmol 2000;84:352-357 PMID- 11004320 TI - Supplemental perioperative oxygen to reduce the incidence of surgical-wound infection. Grief R, Akca O, horn E-P, kurz A, sessler DI* for the outcomes research group. N engl J med 2000;342:161-167 PMID- 11004319 TI - Black diaphragm aniridia intraocular lens for congenital aniridia: long-term follow-up. Reinhard T* engelhardt S, sundmacher R. J cataract refract surg 2000;26:375-381 PMID- 11004322 TI - A prospective search for ocular lesions in hospitalized patients with significant bacteremia. Bouza E* cobo-soriano R, Rodriquez-Creixems M, Munoz P, Suarez-leoz M, Cortes C. Clin infect dis 2000;30:306-312 PMID- 11004321 TI - The elusive causes of keratoconus: a working hypothesis. Kenny MC* brown DJ, rajeev B. CLAO J. 2000;26: 10-13 PMID- 11004323 TI - Risk factors for cortical, nuclear, and posterior subcapsular cataracts: the POLA study. Delcourt C* cristol J-P, tessier F, Leger CL, michel F, papoz L, and the POLA study group. Am J epidemiol 2000;151:497-504 PMID- 11004324 TI - Toxicity of preserved and unpreserved antiglaucoma topical drugs in an in vitro model of conjunctival cells. De saint jean M, debbasch C, brignole F, rat P, warnet J-M, baudouin C.* curr eye res 2000;20:85-94 PMID- 11004325 TI - Faith, hope, and ant piles. PMID- 11004326 TI - Intermittent and continuous ceftazidime infusion for critically ill trauma patients. AB - BACKGROUND: The adequacy of intermittent and continuous infusion ceftazidime for the treatment of nosocomial pneumonia in critically ill trauma patients was assessed by analyzing ceftazidime pharmacokinetics in relation to the minimum inhibitory concentration (MIC) and treatment outcome. METHODS: Serial blood samples were obtained during ceftazidime therapy in 31 trauma patients. Ceftazidime pharmacokinetics were compared with that of previously studied healthy volunteers. Ceftazidime pharmacokinetics were analyzed according to the time above the MIC and treatment outcome. RESULTS: Critically ill trauma patients had a significantly increased volume of distribution and clearance (0.32 +/- 0.14 L/kg and 2.35 +/- 0.89 mL. min(-1). kg(-1), respectively) compared with healthy volunteers (0.21 +/- 0.03 and 1.58 +/- 0.23 mL. min(-1). kg(-1)). The time above the MIC was >/=92% of the dosing interval for all patients and treatment outcomes were similar between the two treatment groups. CONCLUSIONS: Ceftazidime pharmacokinetics are significantly altered in critically ill trauma patients. Both intermittent and continuous ceftazidime regimens were equally effective for the treatment of nosocomial pneumonia caused by less virulent bacteria. PMID- 11004327 TI - Cost analysis of breast conservation surgery compared with modified radical mastectomy with and without reconstruction. AB - BACKGROUND: Breast conservation surgery (BCS), consisting of lumpectomy, axillary lymph node dissection, and radiation therapy, is as effective as modified radical mastectomy (MRM) for the treatment of early stage breast cancer. The costs of these treatment options have not been adequately addressed in the current era of increasing utilization of BCS and breast reconstruction. The purpose of this study is to determine differences in treatment costs among BCS, MRM alone, and MRM with reconstruction. METHODS: Patients with stage I and II breast cancer receiving inpatient treatment at a private university-affiliated hospital between January 1996 and July 1997 were analyzed (n = 230). Charges were determined as follows: inpatient and radiotherapy charges from the hospital billing department, surgeon fees from group practice billing codes, and radiotherapy physician fees from the radiation oncology group practice. Inpatient length of stay was obtained from hospital medical records. RESULTS: Average hospital inpatient charge for BCS was $4,748 (n = 74), $6,280 for MRM alone (P <0.001, n = 132), and $11,946 for MRM with reconstruction (P <0.001, n = 24). Surgeons' fees for BCS were $2,840, $3,500 for MRM alone, and $10,774 for MRM with reconstruction. The average radiotherapy charge was $18,742. Average length of stay was 1.03 days for BCS, 2.44 days for MRM alone (P <0.001), and 3.71 days for MRM with reconstruction (P <0. 001). Average total cost of BCS ($26,330) was significantly greater than the average total cost of either MRM alone ($9,780, P <0.001) or MRM with reconstruction ($22,720, P <0.001). CONCLUSIONS: BCS is more expensive than MRM with or without reconstruction. It is the addition of radiotherapy that results in the higher total cost of CS. PMID- 11004328 TI - The correlation of axillary ultrasonography with histologic breast cancer downstaging after induction chemotherapy. AB - BACKGROUND: The goal of this study was to examine the role of ultrasonography in detecting axillary lymph node metastases in stage II breast cancer patients after induction chemotherapy (IC). METHODS: Of 172 consecutive patients with T1-3, N0 1, M0 breast cancer registered in a prospective IC trial, a subset of 130 evaluable patients were chosen, with (1) both physical and ultrasonographic examinations of the axilla before and after IC; (2) exactly four cycles of IC; (3) no presurgical radiation therapy; and (4) an axillary lymph node dissection. RESULTS: Before IC, 32 patients (25%) were negative for axillary involvement by both physical and ultrasonographic examinations. After IC, this number increased to 64 (49%). Of these, 31 (48%) were positive by pathology examination. In most cases, however, the residual tumor was minimal. CONCLUSIONS: Stage II breast cancer patients who were or became node negative by both ultrasonographic and physical examinations after IC had a 48% incidence of nodal metastases. Because the residual tumor was minimal, irradiation may be sufficient for adequate local control of the axilla. PMID- 11004329 TI - Defining a role for endoscopic ultrasound in staging periampullary tumors. AB - BACKGROUND: The goal of the preoperative workup in patients with suspected periampullary carcinoma is to establish the diagnosis with a high degree of certainty. In this study we compared endoscopic ultrasonography (EUS) and computed tomography (CT) scans for the detection of tumor, lymph node metastasis, and vascular invasion in patients with suspected periampullary carcinoma in order to define a role for EUS in the preoperative staging of these patients. METHODS: Thirty-seven consecutive patients received EUS and CT scanning followed by operation for presumed periampullary carcinoma during a 30-month period. Both imaging modalities were reviewed in a blinded fashion and the results compared with pathology and operative reports on all patients. RESULTS: Sensitivity, specificity, positive predictive value, and negative predictive value for tumor detection by EUS were 97%, 33%, 94%, and 50%, respectively, compared with 82%, 66%, 97%, and 25% for CT scan. For lymph nodes the values were 21%, 80%, 57%, and 44%, respectively, for EUS compared with 42%, 73%, 67%, and 50% for CT. For vascular invasion, the values were 20%, 100%, 100%, and 89%, respectively, for EUS, compared with 80%, 87%, 44%, and 96% for CT. CONCLUSIONS: CT is the initial study of choice in patients with suspected periampullary tumors. EUS is superior for detecting tumor and for predicting vascular invasion. Therefore, EUS should be used for patients in whom CT does not detect a mass and for those with an identifiable mass on CT in whom vascular invasion cannot be ruled out. PMID- 11004330 TI - Utility of fluorine-18-fluorodeoxyglucose positron emission tomography in differentiated thyroid carcinoma with negative radioiodine scans and elevated serum thyroglobulin levels. AB - BACKGROUND: This study aimed to determine the role of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) in the follow-up of patients who underwent total thyroidectomy and iodine-131 ((131)I) ablation therapy for differentiated thyroid cancer and presented increased thyroglobulin levels with negative (131)I and thallium-201 ((201)Tl) scans. METHODS: Two patients with follicular carcinoma and eight with papillary tumors underwent total thyroidectomy and (131)I therapy until the (131)I scan was negative. (131)I and (201)Tl scans were performed with negative results in all cases, while serum thyroglobulin measurements were all positive with negative thyroglobulin autoantibodies. One week after the (131)I scans, all the patients underwent FDG PET whole-body scans. RESULTS: The FDG-PET scan detected in 4 patients, a single focal increase of FDG uptake in one lymph node metastasis (subsequently confirmed histologically); in 1 patient, multiple pathological focal uptakes in brain, neck, and chest; and in 1 patient, two mild focal uptakes in the mediastinum, close to the tracheal branch. In 2 other patients, pathological FDG uptakes in cervical spine and mediastinum were not confirmed by other imaging techniques, and in the 2 remaining patients the scan results were inconclusive. The sensitivity of FDG-PET whole-body scan for detecting metastatic thyroid cancer was 60%. CONCLUSIONS: This study indicates that the FDG-PET whole-body scan is a useful tool in the follow-up of patients with differentiated thyroid cancer, negative (131)I and (201)Tl scans and elevated serum thyroglobulin levels. The FDG-PET scan detects metastatic disease in 60% of patients with differentiated thyroid cancer, enabling surgical therapy to be performed on accessible lesions. PMID- 11004331 TI - Physician leadership is essential to the survival of teaching hospitals. AB - BACKGROUND: Academic medical centers (AMCs) face severe financial constraints because they must now compete directly with private providers that focus exclusively on cost-effective healthcare delivery. Educational and research capacities developed at AMCs have been supported by government and third party payers, but government support is diminishing. Physicians are ill-equipped to respond to market pressures. DATA SOURCES: Analyses of cultural change and restructuring in corporate giants such as Greyhound, IBM and FedEx are relevant to teaching hospitals. To succeed, organizations must flatten hierarchy, empower staff, train leaders, and mobilize intellectual capital. Effective leadership is essential. CONCLUSION: Physicians must educate themselves on forces impacting the AMC, understand changes needed in the structure and processes of AMC governance and acquire competencies for leadership and management if AMCs are to survive and thrive. Surgeons should acquire competencies that will enable them to become leaders in the process of AMC transformation. PMID- 11004332 TI - Visual-spatial abilities in surgical training. AB - As surgery continues to advance, we will need to better understand the role visual-spatial abilities play in the acquisition of technical skills. Many universities have established surgical skills centers with specific curricula to teach residents technical skills as adjuncts to operating room learning. Yet, as educators we do not fully understand the role visual-spatial abilities plays in the acquisition of surgical skills. This paper summarizes the research to date on the relevance of visual-spatial abilities to surgical training. PMID- 11004333 TI - Availability of content information does not improve performance on computerized case simulations. AB - BACKGROUND: A previous study evaluated repeated, serial administrations of computer-based simulations. The data demonstrated an increase in scores across rotations during the academic year, but no difference between scores in successive years. METHODS: The initial study only indirectly assessed the effect of information sharing on measured performance. To directly assess the effect of information transfer, 8 computer-based case simulations were administered over 2 consecutive years to 220 third-year students at the conclusion of 12 surgical clerkship rotations (6 per year). During the second year of administration information regarding content area, in the form of the case stem or introductory lead-in material, was provided to each rotation of students prior to the examination based on a sequential algorithm. RESULTS: The data demonstrate no increase in overall mean score for the examination over the 2 years. Scores were significantly different for 2 of the 8 cases, increasing in the one and decreasing in the other. CONCLUSIONS: The data demonstrate no evidence that prior knowledge of content area influences the scores of successive classes on computerized models of performance assessment. PMID- 11004334 TI - Initial results with a stapled gastrojejunostomy for the laparoscopic isolated roux-en-Y gastric bypass. AB - BACKGROUND: The gastric pouch to jejunum anastomosis is a critical step in the performance of an isolated Roux-en-Y gastric bypass. When performing this procedure laparoscopically, intracorporeal suturing of the gastric pouch to Roux en-Y jejunum anastomosis is technically demanding, time consuming, and perhaps the most prohibitive part of the operation. We devised a unique, effective, and simple method to perform this anastomosis using an EEA stapler. This report describes this technique and its follow-up in our series of patients undergoing a laparoscopic isolated Roux-en-Y gastric bypass utilizing this technique. METHODS: A prospective analysis was performed identifying the technical success, leak rate, and postoperative incidence of anastomotic stenosis and its management in a consecutive series of patients undergoing a laparoscopic isolated Roux-en-Y gastric bypass with a gastrojejunal anastomosis constructed with a 21-mm or 25-mm EEA stapler. RESULTS: Forty-eight patients underwent laparoscopic isolated Roux en-Y gastric bypass. Mean age was 40.9 years (range 22 to 64) and mean body mass index was 52.3 kg/m(2) (range 31 to 76 kg/m(2)). There were no mortalities. Three patients (6.3%) were converted to an open procedure, but only 1 because of an inability to perform the gastrojejunal anastomosis (short jejunal mesentery). There was 1 leak (2.1%) from the gastrojejunal anastomosis. It was successfully managed nonoperatively. Thirteen patients (27.1%) patients developed an anastomotic stenosis requiring endoscopic balloon dilatation. Seven of the 13 patients required only a single dilatation and have had no recurrence of dysphagia. Six of the 13 patients needed 2 to 4 dilatations, and all are swallowing normally. None have required surgical revision. After 12 months of follow-up, the mean weight loss was 115 pounds and mean decrease in body mass index was 18.5 kg/m(2). CONCLUSIONS: The stapled EEA gastrojejunal anastomosis for the laparoscopic isolated Roux-en-Y gastric bypass is safe and effective. Anastomotic stenosis occurs in approximately one quarter of patients, but it can be managed well with endoscopic balloon dilatation. PMID- 11004335 TI - Inferior head resection of the pancreas for intraductal papillary mucinous tumors. AB - BACKGROUND: Patients with intraductal papillary mucinous tumor have a favorable prognosis after surgical treatment. When this neoplasm is located in the head of the pancreas, resection has conventionally required pancreatoduodenectomy. Although pancreatoduodenectomy can now be performed with a low mortality rate, morbidity still occurs frequently. METHODS: Between November 1982 and January 1999, 38 intraductal papillary mucinous tumors of the pancreas were resected at the Chiba University Hospital. Seven patients (18%) underwent inferior head resection of the pancreas. In this preliminary study, the operative technique is presented, and its efficacy in improvement of quality of life is evaluated. RESULTS: Patients with intraductal papillary mucinous tumor underwent resection with no perioperative mortality. After discharge from hospital, 6 patients who underwent inferior head resection were still alive without recurrent disease after a median follow-up of 3 years. However, 1 patient developed peritoneal dissemination and died 18 months after inferior head resection. Patients had regained 98% of preoperative weight 1 year after inferior head resection. N benzoyl-L-tyrosyl-p-amino-benzoic acid (BT-PABA) excretion test showed the same value before (73%) and after (73%) inferior head resection (n = 7). Pancreatic fistulas occurred more frequently after inferior head resection (38%), but the incidence of major complications was similar between inferior head resection and other types of pancreatic head resection. CONCLUSIONS: Pancreatic function was well preserved, and patients regained 98% of preoperative weight after inferior head resection of the pancreas. The authors concluded that the limited involvement of intraductal papillary mucinous tumors enables the surgeons to perform inferior head resection of the pancreas. PMID- 11004336 TI - Right colonic intussusception. PMID- 11004337 TI - Visceral artery aneurysms as seen in a community hospital. AB - BACKGROUND: Visceral artery aneurysm (VAA) is a rare but important form of vascular pathology. METHODS: A retrospective chart review was performed for all patients treated for VAAs from 1980 to 1998 at the Northern California Kaiser Hospitals. RESULTS: Thirty-one patients with VAAs were treated, 11 men and 20 women with an average age of 60 years. The arteries involved were 15 splenic, 8 hepatic, 5 superior mesenteric branch, 1 left gastric, 1 gastroduodenal, and 1 left colic. Seventy-four percent of patients presented with acute abdominal pain and/or shock secondary to aneurysm rupture. Angioembolization was performed in 9, and 25 were treated surgically. There was no morbidity, and 1 death (3.2%). CONCLUSIONS: Patients with VAAs often present to the community hospital general surgeon as acute abdominal emergencies. Symptomatic VAAs can be managed successfully by simple ligation in the primary care setting. PMID- 11004338 TI - Perfusion and functional anatomy of the splenic remnant supplied by short gastric vessels. AB - BACKGROUND: Surgeons have described many methods to preserve the injured spleen for the maintenance of host defence. The volume and the perfusion of remaining splenic tissue are important for better functional results. Imaging the functioning tissue with radionuclide provides information about the physiology of the concerning organ. METHODS: Thirty rats were separated into three groups: control, explorative laparotomy alone; partial splenectomy, upper part of the spleen supplied by short gastric vessels was preserved after partial resection; and devascularized spleen, the entire spleen was preserved after ligation of splenic artery. The size, functional anatomy, and perfusion status of splenic tissue were assessed by liver-spleen scintigraphy using radiolabeled heat denatured red blood cells. RESULTS: In the partial splenectomy group, splenic radioactivity count, spleen/liver ratio, and radionuclide uptake were mildly reduced, and found to be 87% (P = 0.012), 91% (P = 0.16), and 88% (P <0.001) of the normal spleen, respectively. The area of functional tissue in the upper splenic remnant was 51% (P <0.001) of the normal spleen. In the devascularized spleen group, the radioactivity count, the ratio, and the uptake were 38% (P <10( 6)), 36% (P <10(-6)), and 49% (P <10(-6)) of the normal spleen respectively. The area of functional tissue in the devascularized spleen was calculated as 47% (P <10(-6)) of the normal spleen. CONCLUSIONS: The collateral circulation is insufficient for proper function of the entire spleen after disconnection of the main arterial blood supply. The functional tissue is markedly impaired. On the other hand, the upper part of the spleen is remained well perfused via the short gastric vessels. The collateral circulation is satisfactory for this splenic tissue after reduction of its volume. That size splenic tissue seems to have satisfactory functional ability. Proper functional results mostly depend on the balance between the volume and blood supply of the remaining splenic tissue. PMID- 11004339 TI - Ligating the cystic duct in laparoscopic cholecystectomy. AB - BACKGROUND: In laparoscopic cholecystectomy, bile leak from the cystic duct stump is a recognised postoperative complication. This could be due to dislodgement of the metal clips. Migration of the clip is an added problem. METHODS: Since 1995, 70 patients were included in the study of intracorporeal cystic duct ligation in laparoscopic cholecystectomy. Construction of a ligature is a single handed procedure using a free end of the thread. It is quite different from the other two methods described in the literature. RESULTS: There was not a single case of postoperative morbidity in these patients, but in a retrospective study, postoperative biliary dyspepsia with fatty food was evident in a few patients among those cases in which the liga-clips were applied to the cystic duct. CONCLUSION: The cystic duct ligation with absorbable thread should be a gold standard in laparoscopic cholecystectomy. It reduces the risk of postoperative morbidity. PMID- 11004340 TI - Bilateral Jejuno-mesenteric flap for reconstruction of complicated pharyngoesophageal defect. AB - The surgical management of an infectious and fistulous wound with a pharyngoesophageal tumor is one of the greatest challenges for head and neck and plastic surgeons. The free jejunal transfer has been the standard technique for pharyngoesophageal reconstruction, and the free omental flap has been one of the most reliable methods for reconstructing contaminated wounds. A jejuno-mesenteric flap is suitable for such complicated wounds. Pharyngoesophageal defects are reconstructed by the jejunum, and contaminated and heavily irradiated neck wounds are covered with the mesenteric flaps connected with a revascularized jejunum. The technique described here possesses the advantages of both a free jejunal flap and an omentum flap. Therefore, it is a reliable method for reconstructing the pharyngoesophageal defects of complicated wounds. PMID- 11004341 TI - Laparoscopic colorectal cancer surgery. AB - BACKGROUND: A number of controversies exist in the laparoscopic treatment of colorectal cancer, and thus the technique has so far failed to gain widespread acceptance throughout the United Kingdom. This review aims to discuss these issues in the context of ongoing published trials, assessing both purported advantages and disadvantages. METHODS: The United States National Library of Medicine Medline database, and the Bath Information Data Service (BIDS) were searched using keywords related to laparoscopic colorectal cancer surgery. Recent surgical journals were also reviewed for relevant publications. Attempts have been made to quote only the most recent work from institutions with multiple publications using the same group of patients, in order to present the most coherent picture. The data are presented as randomized controlled trials, nonrandomized controlled studies, and series comprising more than 10 patients. CONCLUSIONS: This review confirms that laparoscopic colorectal cancer surgery is technically feasible. In addition patients lose less blood, have less immunosuppression, and have shorter postoperative ileus, in-patient stay, and require less analgesia. However, concerns still remain as to the development of port-site metastases, the longer operating times, and the overall cost of the equipment. In view of these concerns, the place of laparoscopically assisted colorectal cancer surgery is likely to remain controversial for some years yet. Randomized, controlled trials are as yet too few to provide definitive answers to all these issues. PMID- 11004342 TI - The role of multimodality therapy for resectable esophageal cancer. AB - BACKGROUND: There is an increasing interest in the role of combined therapy to achieve long-term survival for patients with resectable esophageal neoplasms. Surgery provides excellent palliation with relatively low morbidity and mortality rates, but cure remains elusive. MATERIAL AND METHODS: From January 1988 to January 1998, a total of 137 patients met eligibility criteria for a combined multimodal therapy, prospective, nonrandomized protocol of induction chemoradiation therapy followed by surgical resection, based on radiological and endoscopic assessment of the extension (all patients were initially considered to be at clinical stages I to III, locoregional). Consequently, patients with high grade Barrett's dysplasia or any squamous carcinoma in situ (stage 0) and those with distant metastatic disease (stage IV) were excluded. Among this group, 48 operable patients with biopsy-proven esophageal cancer finally entered and completed the protocol and are the sample of the present study. Multivariate logistic regression models were used to identify risk factors for death or recurrence. Actuarial survival was calculated since the beginning of the induction protocol by the Kaplan-Meier method, and comparisons between groups were made by the log-rank test. RESULTS: Mean age was 61.6 (range 45 to 71), and 72.9% were male. The majority of the tumors (70.8%) were located at the lower third/cardia and as many as 18.8% were adenocarcinoma. After a mean of 7.5 weeks (range 5 to 12) after the completion of the induction phase, 68.7% underwent a transthoracic esophagectomy and 31.3% a transhiatal esophagectomy. The in hospital mortality rate was 10.4% (5 patients). A complete response (no evidence of tumor within the specimen: pT0) was achieved in 25% (12 patients). After a mean follow-up of 20.2 months, mean survival for the entire group was 18.2 months (95% confidence interval 14 to 22). At the end of the study, 25% (12) remained alive. Actuarial survival rates at 12, 23, and 37 months were 56.2%, 36.9%, and 21.9%, respectively. CONCLUSIONS: Esophageal resection after induction therapy seems to be related to a slightly higher mortality rate compared with historical series, and for this reason, neoadjuvant therapy must be considered still experimental. However, no statistical significant difference in survival is showed in those cases with complete pathological response (pT0). Factors influencing survival are recurrence and age. Surgery alone remains the standard therapy for esophageal cancer. PMID- 11004344 TI - A prognostic score for gastric cancer. AB - BACKGROUND: Scoring systems are generally used for predicting prognosis in the intensive care unit, but there is no score being used frequently for predicting prognosis in gastric cancer. The aim of this study was to develop a prognostic score for gastric cancer patients. METHODS: Demographic, clinical, laboratory, radiologic, histopathologic data, and operative findings for 128 patients who had curative or palliative resection for gastric cancer were analyzed for their effect on overall and disease-free survival. Ten variables-invasion depth of tumor, node status (American Joint Committed on Cancer, 1992), metastasis, node status (Union Internationale Contre le Cancer, 1997), metastatic lymph node ratio, resectability, tumor location, extent of lymphadenectomy, Borrmann type, Lauren type-that have independent significant effect or borderline significance on both overall and disease-free survival according to multivariate analysis were chosen. Coefficients were calculated for these variables by using Cox regression analysis, and thus the Prognostic Score for Gastric Cancer (PSGC) was designed. All patients were scored using the PSGC and also staged clinically (AJCC 1992) and histopathologically (AJCC 1992 and UICC 1997). RESULTS: Patients were grouped according to their scores: group 1, patients with scores 20 to 50 (probability of 5-year overall survival 50% to 95%); group 2, patients with scores 51 to 80 (probability of 5-year overall survival 10% to 50%); and group 3, patients with scores 81 and higher (probability of 5-year overall survival <10%). Overall survival and disease-free survival decreased significantly with increasing scores. The association of PSGC and staging systems with survival was analyzed by stepwise logistic regression and Cox regression analyses. PSGC was proved to have the most significant association with overall and disease-free survival. CONCLUSIONS: Inclusion of more variables in PSGC seems to make it superior than staging. It is easy to adapt PSGC to different patient populations, which may make it accepted as a practical and useful scoring system in clinical practice. PMID- 11004343 TI - Is MIB-1 proliferation index a predictor for response to neoadjuvant therapy in patients with esophageal cancer? AB - BACKGROUND: The overall survival rate for patients with an esophageal cancer remains poor. As a consequence, preoperative chemoradiation was introduced for patients with tumor stage T >1 M0 regardless of tumor histology or localization. However, factors predicting response to this therapy pretherapeutically are largely unknown. METHODS: Clinical results of preoperative chemoradiation were investigated. The rates of proliferation and apoptosis were determined in pretherapeutic tumor samples and correlated with tumor response and long-term survival after surgery. RESULTS: A complete tumor response due to chemoradiation (n = 42; cervically localized tumors excluded) was achieved in 11 patients (26%) after resection. Five-year survival rate was significantly improved in these patients compared with those who did not respond to chemoradiation (48% versus 5.5%; P = 0.003). Chemoradiation was performed without benefit in 43%. Perioperative hospital mortality rate was 14.3% in all patients. No correlation of apoptosis with response to chemoradiation or postoperative long-term survival was observed. However, there was a clear correlation between the proliferation rate as determined by MIB-1 immunohistology. Five-year survival rate of patients with a proliferation index (PI) >/=39% was 38% compared with 0% in tumors with a PI <39%. Tumors with a PI >/=39% responded to chemoradiation in 71.4%, but 100% of tumors with a PI <39% did not. Mean survival time of these patients was 33 months and 11 months, respectively (P = 0.015). CONCLUSIONS: The results indicate that the PI may be used for stratification of patients treatment prior surgery. However, these results need further validation in larger patient numbers in the search for factors indicating response pretherapeutically to preoperative chemoradiation in esophageal cancer. PMID- 11004345 TI - Continued follow-up of pregnancy outcomes in diethylstilbestrol-exposed offspring. AB - OBJECTIVE: To evaluate long-term pregnancy experiences of women exposed to diethylstilbestrol (DES) in utero compared with unexposed women. METHODS: This study was based on diethylstilbestrol-exposed daughters, the National Collaborative Diethylstylbistrol Adenosis cohort and the Chicago cohort, and their respective nonexposed comparison groups. Subjects who could be traced were sent a detailed questionnaire in 1994 that contained questions on health history, including information on pregnancies and their outcomes. We reviewed 3373 questionnaires from exposed daughters and 1036 questionnaires from unexposed women. RESULTS: The response rate was 88% among exposed and unexposed women. Diethylstilbestrol-exposed women were less likely than unexposed women to have had full-term live births and more likely to have had premature births, spontaneous pregnancy losses, or ectopic pregnancies. Full-term infants were delivered in the first pregnancies of 84.5% of unexposed women compared with 64. 1% of exposed women identified by record review (relative risk [RR] 0.76, confidence interval [CI] 0.72, 0.80). Preterm delivery of first births occurred in 4.1% of unexposed compared with 11.5% of exposed women, and ectopic pregnancies in 0.77% of unexposed compared with 4.2% of exposed women. Spontaneous abortion was reported in 19.2% of DES-exposed women compared with 10.3% in control women (RR 2.00, CI 1.54, 2.60). According to complete pregnancy histories (many women had more than one pregnancy), preterm births were more common in DES-exposed women (19.4% exposed versus 7.5% unexposed (RR 2.93 CI 2.23, 3.86). Second-trimester spontaneous pregnancy losses were more common in DES-exposed women (6.3% versus 1.6%; RR 4.25, CI 2.36, 7.66). More first trimester spontaneous abortions occurred in DES-exposed women than in controls (RR 1.31, CI 1.13, 1.53), and DES-exposed women had at least one ectopic pregnancy more often than unexposed women (RR 3.84, CI 2.26, 6.54). CONCLUSION: Pregnancy outcomes in DES-exposed women were worse than those in unexposed women. PMID- 11004346 TI - Serial salivary estriol to detect an increased risk of preterm birth. AB - OBJECTIVE: To evaluate serial measurements of salivary estriol (E3) to detect increased risk of spontaneous preterm labor and preterm birth. METHODS: A masked, prospective, multicenter trial of 956 women with singleton pregnancies was completed at eight United States medical centers. Saliva was collected weekly, beginning at the 22nd week of gestation until birth, and tested for unconjugated E3 by enzyme-linked immunosorbent assay. Women were separated into high-risk and low-risk groups using the Creasy scoring system. RESULTS: A single, positive (at or above 2.1 ng/mL) salivary E3 test predicted an increased risk of spontaneous preterm labor and delivery in the total population (relative risk [RR] 4.0, P <.005), in the low-risk population (RR 4.0, P < or =.05), and in the high-risk population (RR 3.4, P =.05). Two consecutive positive tests significantly increased the RR in all study groups, with a dramatic improvement in test specificity and positive predictive value but only a modest decrease in sensitivity. In women who presented with symptomatic preterm labor, salivary E3 identified 61% of those who delivered within 2 weeks, using a threshold of 1.4 ng/mL. CONCLUSION: Elevated salivary E3 is associated with increased risk of preterm birth in asymptomatic women and symptomatic women who present for evaluation of preterm labor. PMID- 11004347 TI - Group B streptococcal colonization and serotype-specific immunity in pregnant women at delivery. AB - OBJECTIVE: To describe the relationship between serum concentration of group B streptococcal capsular polysaccharide-specific immunoglobulin (Ig) G, colonization status, race or ethnicity, and age in pregnant women. METHODS: Pregnant women (n = 3307) were enrolled from geographically and ethnically diverse populations. At the time of admission for delivery, swabs of the lower vagina and rectum were obtained for isolation of group B streptococci. In a subset of women whose sera were available, capsular polysaccharide-specific IgG concentrations were quantified by serotype-specific (Ia, Ib, II, III, and V) enzyme-linked immunosorbent assay and compared by group B streptococcal colonization status. RESULTS: Group B streptococcal colonization was detected in 856 women (26%), and the rate was significantly higher among black women (37%) than in other racial or ethnic groups (odds ratio 1.7, 95% confidence interval 1.4, 2.1). Colonization status did not differ by study site or age. Colonization with serotypes Ia, II, III, or V was associated with significantly higher serum concentrations of IgG specific for the capsular polysaccharide of the colonizing serotype compared with noncolonization. However, 48% of colonized women had low capsular polysaccharide-specific IgG levels (less than 0.5 microg/mL) in their delivery sera. Colonized teenagers had the lowest median concentration. CONCLUSION: Colonization with group B streptococcus can elicit a systemic immune response, with a cumulative increase in the prevalence of capsular polysaccharide specific IgG with increasing age. Conversely, low antibody levels in colonized teenagers might account in part for the reported increased risk of group B streptococcal disease in neonates born to these patients. PMID- 11004348 TI - Wet smear compared with gram stain diagnosis of bacterial vaginosis in asymptomatic pregnant women. AB - OBJECTIVE: To compare wet smear and Gram stain diagnoses of bacterial vaginosis among asymptomatic pregnant women. METHODS: Between November 1, 1996 and December 31, 1997, asymptomatic women who initiated prenatal care in our obstetric clinics were invited to participate. Exclusion criteria included antimicrobial use within 2 weeks, cervical cerclage, vaginal bleeding, placenta previa, spermicide use, douching, or intercourse within 8 hours. Clinical diagnosis that required two of three positive criteria for bacterial vaginosis (vaginal pH, whiff test, and clue cells on wet smear) was compared with Gram stain diagnosis (Bacterial vaginosis score 7-10 by Nugent criteria). RESULTS: Population characteristics (n = 69) included an average (+/- standard deviation [SD]) maternal age of 27. 3 +/- 6.6 years, 26 nulliparas (38%), 28 black women (41%), 23 white women (38%), 15 Hispanic women (22%), and three Asian women (4%). The mean (+/-SD) gestational age at entry was 15.6 +/- 7.6 weeks. Twenty-seven percent (18 of 67) of the study population was diagnosed with bacterial vaginosis by definitive Gram stain. Two slides were lost or were of poor quality and not included. Using Gram stain diagnosis of bacterial vaginosis as the standard, clinical diagnosis had sensitivity of 56% (95% confidence interval [CI] 32%, 78%), a specificity of 96% (95% CI 90%, 100%), a positive predictive value of 83%, and a negative predictive value of 85%. CONCLUSION: In asymptomatic pregnant women, bacterial vaginosis can be diagnosed reliably by Gram stain. PMID- 11004349 TI - Relation between serum uric acid and plasma adenosine levels in twin pregnancies. AB - OBJECTIVE: To examine the relationship between plasma adenosine and serum uric acid levels in women with singleton and twin pregnancies. METHODS: We sampled maternal arterial blood and measured serum uric acid and plasma adenosine levels in 22 singleton pregnancies and nine twin pregnancies at 33 to 38 weeks' gestation. RESULTS: The average plasma adenosine levels were 0.31 +/- 0.12 micromol/L in the singleton pregnancy group and 0.45 +/- 0.09 micromol/L in the twin pregnancy group (P <.001). The mean serum uric acid level in women with twin pregnancy was 5.7 +/- 0.44 mg/dL which was higher than that in the singleton pregnant women (4.4 +/- 0.69 mg/dL, P <.001). Positive correlations were found between serum uric acid and plasma adenosine levels in both the singleton (r(2) = 0.54, P <.001) and the twin pregnancy groups (r(2) = 0.65, P =.009). Moreover, there was also a significant correlation between serum uric acid and plasma adenosine levels overall (r(2) = 0.66, P <.001). CONCLUSION: Our results suggest that higher adenosine levels are a contributing source of hyperuricemia in twin pregnancies. PMID- 11004350 TI - Procedure-related miscarriages and Down syndrome-affected births: implications for prenatal testing based on women's preferences. AB - OBJECTIVE: To determine how pregnant women of varying ages, races, ethnicities, and socioeconomic backgrounds value procedure-related miscarriage and Down syndrome-affected birth. METHODS: We studied cross-sectionally 534 sociodemographically diverse pregnant women who sought care at obstetric clinics and practices throughout the San Francisco Bay area. Preferences for procedure related miscarriage and the birth of an infant affected by Down syndrome were assessed using the time trade-off and standard gamble metrics. Because current guidelines assume that procedure-related miscarriage and Down syndrome-affected birth are valued equally, we calculated the difference in preference scores for those two outcomes. We also collected detailed information on demographics, attitudes, and beliefs. RESULTS: On average, procedure-related miscarriage was preferable to Down syndrome-affected birth, as evidenced by positive differences in preference scores for them (time trade-off difference: mean = 0.09, median = 0.06; standard gamble difference: mean = 0.11, median = 0.02; P <.001 for both, one-sample sign test). There was substantial subject-to-subject variation in preferences that correlated strongly with attitudes about miscarriage, Down syndrome, and diagnostic testing. CONCLUSION: Pregnant women tend to find the prospect of a Down syndrome-affected birth more burdensome than a procedure related miscarriage, calling into question the equal risk threshold for prenatal diagnosis. Individual preferences for those outcomes varied profoundly. Current guidelines do not appropriately consider individual preferences in lower-risk women, and the process for developing prenatal testing guidelines should be reconsidered to better reflect individual values. PMID- 11004351 TI - Complications of hysteroscopic surgery: predicting patients at risk. AB - OBJECTIVE: To determine the frequency of operative complications and whether they can be predicted by specific patient characteristics or type of hysteroscopic procedure. METHODS: We collected demographic and medical history information on 925 women who had hysteroscopies from 1995 through 1996. We compared differences in rates of operative complications of specific hysteroscopic procedures. Operative complications were defined as uterine perforation, excessive glycine absorption (1 L or more), hyponatremia, hemorrhage (500 mL or more), bowel or bladder injury, inability to dilate the cervix, and procedure-related hospital admissions. RESULTS: Operative complications occurred in 25 (2.7%) of 925 hysteroscopies. Excessive fluid absorption was the most frequent complication. Hysteroscopic myomectomy and resection of uterine septum were associated with greater odds of complications (odds ratio [OR] 7.4, 95% confidence interval [CI] 3.3, 16.6 and OR 4.0, 95% CI 0.9, 19.6, respectively). Hysteroscopic polypectomy and endometrial ablation were associated with lower odds of complications (OR 0.1, 95% CI 0.0, 0.7 and OR 0.4, 95% CI 0.1, 3.3, respectively). Hysteroscopies done by reproductive endocrinologists and preoperative GnRH agonist therapy were associated with 4-7 times higher odds for operative complications. CONCLUSION: Complications during hysteroscopic surgery are rare. Among hysteroscopic procedures, myomectomies and resections of uterine septa have significantly higher rates of complications, especially excessive fluid absorption. Meticulous fluid management might limit the number of serious complications of these higher risk procedures. PMID- 11004352 TI - Eicosapentaenoic acid effect on hyperlipidemia in menopausal Japanese women. The Niigata Epadel Study Group. AB - OBJECTIVE: To assess the efficacy and safety of eicosapentaenoic acid for the treatment of hyperlipidemia in symptomatic menopausal Japanese women. METHODS: We performed a prospective observational 48-week study in hyperlipidemic menopausal women. We randomly assigned 141 women, whose levels of serum total cholesterol were 220 to 280 mg/dL or whose serum triglycerides were 150 to 400 mg/dL at baseline to groups treated with estriol (E3) 2 mg daily (control group, n = 72) or ethyl icosapentate 1800 mg daily and E3 2 mg (eicosapentaenoic acid group, n = 69). RESULTS: Serum levels of total cholesterol decreased significantly from 249.4 to 238.6 mg/dL (-4.3%, P =.003) in the control group and from 252.3 to 234.0 mg/dL (-7.3%, P =.001) in the study group at week 48 in the women whose total cholesterol was not less than 220 mg/dL at baseline. Serum levels of triglycerides decreased significantly from 194.5 to 141.5 mg/dL (-27. 2%, P =.001) in the study group but increased slightly from 192.9 to 207.4 mg/dL (+7.5%) in the control group at week 48 in the women whose level of triglycerides was not less than 150 mg/dL. There were significant differences between these two groups at weeks 12, 24, and 48. Serum levels of total cholesterol and triglycerides were significantly decreased at week 48 in the study group regardless of whether the women were obese. There were no severe adverse effects. CONCLUSION: Combination therapy with eicosapentaenoic acid and E3 was effective and safe for menopausal women with hypertriglyceridemia. PMID- 11004353 TI - Safety of extraovular catheter insertion for second-trimester abortion. AB - OBJECTIVE: To evaluate the efficacy and safety of second-trimester abortions using transcervical catheter insertion and extraovular prostaglandin (PG) administration. METHODS: Ninety women admitted for terminations of pregnancy at 17-24 weeks' gestation had transcervical catheters inserted and extraovular PGE(2) administered. Success rates were recorded, measured by induction of abortion within 24 hours, need for a complement uterine curettage, and complications. RESULTS: The technique induced abortion in 67 women (74.4%). The induction-to-abortion median interval was 12 hours (7 and 22 hours, fifth and 95th percentiles, respectively). Thirty-seven women needed uterine curettages because of incomplete abortions or excessive uterine bleeding after fetal and placenta expulsion. One woman had shivering, weakness, and nausea attributed to systemic absorption of PG, and nine women developed systemic inflammatory response syndrome associated with transcervical catheter insertion. Two of those women had septic shock, one of whom deteriorated to a life-threatening situation. CONCLUSION: Transcervical catheter insertion for extraovular PG administration is effective for inducing second-trimester abortions. Although the method is considered safe, with generally few mild, treatable complications, we observed a high rate of systemic inflammatory response syndrome, bacteremia, and sepsis caused by transcervical catheter insertion before PG administration. A reconsideration of this method's safety is warranted. PMID- 11004354 TI - Prelabor rupture of the membranes at term: expectant management at home or in hospital? The TermPROM Study Group. AB - OBJECTIVE: To determine whether adverse effects of expectant management for premature rupture of membranes (PROM) at term and patient satisfaction were greater if women were managed at home rather than in a hospital. METHODS: We undertook a secondary analysis of data from the International TermPROM Study for women managed expectantly at home or in a hospital. Using multiple logistic regression analyses, we determined the effect of home and hospital management and controlled for differences in baseline characteristics, in measures of maternal and neonatal infections and rates of cesarean. RESULTS: Six hundred fifty-three women (39.1%) were managed at home, and 1017 (60.9%) in a hospital. Management at home, compared with in a hospital, increased risk of nulliparas needing antibiotics before delivery (odds ratio [OR] 1.52 95% confidence interval [CI] 1.04, 2.24, P =.03), those not colonized with group B streptococcus having cesareans (OR 1.48 95% CI 1.03, 2. 14, P =.04), and neonatal infections (OR 1.97 95% CI 1.00, 3.90, P =. 05). More multiparas managed at home said they would participate in the study again (OR 1.80 95% CI 1.27, 2.54, P <.001). CONCLUSION: Expectant management at home, rather than in a hospital, might increase the likelihood of some adverse outcomes. PMID- 11004355 TI - Membrane sweeping in conjunction with labor induction. AB - OBJECTIVE: To determine whether cervical membrane sweeping (stripping) during induction of labor is beneficial. METHODS: We compared outcomes of labor after induction in pregnant women at term in a randomized trial. Women were assigned to having their membranes swept or not during induction. Outcome measures included duration of labor, maximum dose of oxytocin used, induction-labor interval, and mode of delivery. RESULTS: We recruited 130 nulliparas (64 sweep, 66 nonsweep) and 118 multiparas (60 sweep, 58 nonsweep). Among nulliparas who received intravaginal prostaglandin (PG) E(2) and oxytocin, those who had simultaneous sweeping had significantly shorter mean (+/- standard error of mean) induction labor interval (13.6 +/- 1.4 versus 17.3 +/- 1.2 hours, P =.048), lower mean maximum dose of oxytocin (6.8 +/- 0.8 versus 10.35 +/- 1.1 mU/minute, P =.01), and increased normal delivery rates (vaginal delivery 83. 3% versus 58.2%, P =.01). Sweeping also had a favorable effect on nulliparas who received oxytocin alone (mean induction-labor interval 5.8 +/- 3.1 versus 11.2 +/- 3.6 hours, P =.04; mean maximum dose 8.8 +/- 1.3 versus 16.3 +/- 1.9 mU/min, P =.01). Those differences were limited to women with unfavorable cervices. There were no differences in any outcome measures in multiparous women. CONCLUSION: Sweeping of the membranes during induction of labor had a beneficial effect on labor and delivery, which appeared to be limited to nulliparas with unfavorable cervices who needed cervical priming with PGE(2). PMID- 11004356 TI - Mifepristone for preinduction cervical ripening beyond 41 weeks' gestation: a randomized controlled trial. AB - OBJECTIVE: To compare the effect of mifepristone with placebo on cervical ripening before labor induction in prolonged pregnancies. METHODS: One hundred eighty women with pregnancies beyond 41 weeks and undilated, uneffaced cervices were assigned randomly to receive mifepristone 200 mg or placebo and observed for 24 hours. We then gave intravaginal misoprostol 25 microg every 4 hours or intravenous oxytocin. We expected 60% of placebo-treated and 80% of mifepristone treated women to deliver vaginally within 48 hours. RESULTS: Among 180 subjects, 97 received mifepristone and 83 received placebo. The mean interval (+/- standard deviation [SD]) from start of induction to delivery was 2209 +/- 698 minutes for mifepristone-treated subjects and 2671 +/- 884 minutes for placebo-treated subjects (P <.001, log-transformed data). Twelve (13. 6%) mifepristone-treated women and seven (10.8%) placebo-treated women delivered vaginally on day 1 (P =.60). After 24 hours, the median Bishop score for both groups was 3 (0-11) (P =.51). One hundred thirty-one subjects required misoprostol, 65 (67.0%) were mifepristone-treated women, and 66 (79.5%) placebo-treated women (P =.06). The median (range) oxytocin dose was 871.5 (0-22,174) mU for mifepristone-treated women and 2021.0 (0-24,750) mU for placebo-treated women (P =.02). Seventy-seven (87.5%) mifepristone-treated women and 46 (70.8%) placebo-treated women delivered vaginally 48 hours after the start of treatment (P =.01). There were nine cesareans in the mifepristone group and 18 in the placebo group (P =.02). More nonreassuring fetal heart rate patterns and uterine contractile abnormalities occurred in mifepristone-treated subjects. There were no statistically significant differences in neonatal outcomes between groups. CONCLUSION: Mifepristone had a modest effect on cervical ripening when given 24 hours before labor induction, appearing to reduce the need for misoprostol and oxytocin compared with placebo. PMID- 11004357 TI - Randomized comparison of glyceryl trinitrate and prostaglandin E2 for cervical ripening at term. AB - OBJECTIVE: To estimate the adverse effects of glyceryl trinitrate compared with prostaglandin (PG) E2 vaginal tablet for cervical ripening in term pregnancy. METHODS: One hundred ten women with term pregnancies referred for induction of labor with Bishop scores of 6 or less were randomly assigned to receive a 500 microg glyceryl trinitrate tablet vaginally (n = 54) or a 3-mg PGE2 tablet vaginally (n = 56), every 6 hours for maximum of two doses. Subjects were sent to the labor ward for amniotomy or oxytocin if their Bishop scores were more than 6 or their cervices were not ripe 24 hours after treatment. Adverse effects, changes in the Bishop scores, progress, and outcomes of labor were assessed. RESULTS: Glyceryl trinitrate was associated with fewer episodes of uterine tachysystole (0% versus 9%; P =.02). The median Bishop score after 12 hours was lower in women given glyceryl trinitrate compared with those given PGE2. Adverse effects, including headache and palpitations, were more frequent with glyceryl trinitrate than with PGE2. The cesarean rate was not significantly different between groups. CONCLUSION: Cervical ripening with glyceryl trinitrate resulted in fewer episodes of tachysystole, but there were significantly more minor side effects. It can be used for cervical ripening at term, but it was not as effective as PGE2. PMID- 11004358 TI - Ultrasonography for cervical length measurement: agreement between transvaginal and translabial techniques. AB - OBJECTIVE: To assess agreement between transvaginal and translabial ultrasonography for measuring cervical length in the second and third trimesters. METHODS: Eighty-four obstetric patients at 14-40 weeks' gestation were enrolled. Three images of the cervix were obtained by translabial (3.5-MHz curvilinear transducer) and transvaginal (6.5-MHz endovaginal probe) ultrasonography. The mean cervical length obtained by using each method was used for data analysis. The McNemar chi(2) test was used to assess the difference between techniques in their ability to obtain a measurement. The paired t-test was used to evaluate the differences between the measurements. The mean difference and SD for the differences were used to calculate the limits of agreement. An acceptable difference was defined as less than 0.5 cm. RESULTS: Cervical length measurements were obtained in 84 patients (100%) by using the transvaginal technique and in 80 patients (95%) by using the translabial technique (P =.1). Eighty patients had both transvaginal and translabial measurements for comparison. The mean difference in cervical length was 0.37 cm (P <.001; 95% confidence interval [CI] 0. 21, 0.52). The upper and lower limits of agreement were 1.75 cm (95% CI 1.48, 2.02) and -1.01 cm (95% CI -0.74, -1.28), respectively. The differences between the two measurements were within these limits 95% of the time. These limits of agreement were greater than the acceptable difference of 0.5 cm. CONCLUSION: Transvaginal and translabial techniques should not be used interchangeably for clinical assessment of cervical length because agreement between the methods is not within an acceptable range. PMID- 11004359 TI - One-stage screening for pregnancy complications by color Doppler assessment of the uterine arteries at 23 weeks' gestation. AB - OBJECTIVE: To estimate the value of screening for preeclampsia and fetal growth restriction by performing color Doppler assessment of uterine arteries at 23 weeks' gestation in predicting adverse pregnancy outcome. METHODS: Women with singleton pregnancies who attended routine ultrasonography at 23 weeks had color Doppler uterine artery imaging. Bilateral uterine artery notches were noted and left and right uterine artery pulsatility indices (PI) were measured. A mean PI of more than 1.45 was considered increased. Screening characteristics for predicting preeclampsia and delivery of small-for-gestational-age infants were calculated. RESULTS: Of 1757 pregnancies, increased PI was present in 89 (5.1%) and bilateral notches were noted in 77 (4.4%). Twenty-three of 65 women (35.3%; 95% confidence interval [CI] 23.9, 48.2) had increased PI and later developed preeclampsia, and 8 of 10 (80%; 95% CI 44.4, 97. 5) with preeclampsia required delivery before 34 weeks. The respective values for women with bilateral notches were 21 of 65 (32. 3%; 95% CI 21.2, 45.1) and 8 of 10 (80%; 95% CI 44.4, 97.5). The sensitivity of increased PI was 30 of 143 (21%; 95% CI 14.6, 28.6) for delivery of an infant with birth weight below the tenth percentile and 7 of 10 (70% 95% CI 34.8,93.3) for birth weight below the tenth percentile delivered before 34 weeks. The respective values for bilateral notches were 19 of 143 (13.3%; 95% CI 8.2, 20) and 5 of 10 (50%; 95% CI 18.7, 81.3). CONCLUSION: A one stage color Doppler screening program at 23 weeks identified most women who subsequently developed serious complications of impaired placentation associated with delivery before 34 weeks. The screening results were similar when the high risk group was defined as women with increased PI or bilateral notches. PMID- 11004360 TI - Maximal exercise testing in late gestation: fetal responses. AB - OBJECTIVE: To determine the fetal response to and safety of maximal maternal exercise in the third trimester. METHODS: Twenty-three active women with uncomplicated pregnancies (singleton gestations) underwent maximal exercise testing in late gestation using a progressive maximal cycle ergometer protocol. Fetal heart rate (FHR) responses were monitored and classified using National Institute of Child Health and Human Development guidelines. Statistical analyses involved use of the Student t test, repeated measures analysis of variance with Tukey-Kramer multiple comparisons posttest, and the chi(2) test. RESULTS: There was an increase in baseline FHR in the 20-minute posttest period compared with the 20-minute pretest period. There were significantly fewer accelerations in the second posttest 10-minute segment compared with the second pretest 10-minute segment. Variability was reduced in both posttest periods compared with the first 10-minute pretest period. Time to reactivity increased after testing. Mild tachycardia was noted in two tracings and bradycardia occurred in a fetus with previously undiagnosed growth restriction. There were no abnormal neonatal outcomes. CONCLUSION: Maximal exercise testing in late gestation led to minimal changes in FHR. Fetal bradycardiac responses were not seen in appropriate for gestational age fetuses, suggesting that brief maximal maternal exertion for research or diagnostic purposes is safe in this group. PMID- 11004361 TI - Randomized trial of intermittent or continuous amnioinfusion for variable decelerations. AB - OBJECTIVE: To determine whether continuous or intermittent bolus amnioinfusion is more effective in relieving variable decelerations. METHODS: Patients with repetitive variable decelerations were randomized to an intermittent bolus or continuous amnioinfusion. The intermittent bolus infusion group received boluses of 500 mL of normal saline, each over 30 minutes, with boluses repeated if variable decelerations recurred. The continuous infusion group received a bolus infusion of 500 mL of normal saline over 30 minutes and then 3 mL per minute until delivery occurred. The ability of the amnioinfusion to abolish variable decelerations was analyzed, as were maternal demographic and pregnancy outcome variables. Power analysis indicated that 64 patients would be required. RESULTS: Thirty-five patients were randomized to intermittent infusion and 30 to continuous infusion. There were no differences between groups in terms of maternal demographics, gestational age, delivery mode, neonatal outcome, median time to resolution of variable decelerations, or the number of times variable decelerations recurred. The median volume infused in the intermittent infusion group (500 mL) was significantly less than that in the continuous infusion group (905 mL, P =.003). CONCLUSION: Intermittent bolus amnioinfusion is as effective as continuous infusion in relieving variable decelerations in labor. Further investigation is necessary to determine whether either of these techniques is associated with increased occurrence of rare complications such as cord prolapse or uterine rupture. PMID- 11004362 TI - Management strategy for fetal tachycardia. AB - OBJECTIVE: To develop a management strategy for fetal tachycardia. METHODS: Forty four fetuses (20-40 weeks' gestation) with nonsinus tachycardia were divided into three groups based on duration of tachycardia and degree of heart failure. Fetuses with intermittent tachycardia were treated expectantly. Fetuses with sustained tachycardia were treated with transplacental antiarrhythmic agents alone if heart failure was mild to moderate, and with direct intramuscular therapy if heart failure was severe. Degree of heart failure was determined by echocardiographic variables of ventricular function, atrioventricular valve insufficiency, and hydrops. Fetal well-being and response to treatment were evaluated by daily heart rate surveillance and frequent fetal echocardiograms and ultrasounds. RESULTS: Fifteen fetuses with intermittent tachycardia (n = 15, group 1) did not progress to sustained tachycardia or heart failure. Fetuses with sustained tachycardia and mild-to-moderate heart failure (n = 14, group 2) were cardioverted or rate controlled with transplacental agents (n = 9); three term fetuses were delivered electively without treatment and two progressed to severe heart failure and were treated in group 3. Seventeen fetuses (15 initially, two progressing) with severe heart failure were cardioverted (in 0. 25-21 days; mean 4.3 days) with fetal intramuscular plus transplacental antiarrhythmic therapy (group 3). Overall, 43 of 44 fetuses were delivered at 32 to 41 (mean 37) weeks with minimal morbidity and a mortality rate of 2.2% (95% confidence interval 0. 06%, 12.0%). CONCLUSION: Perinatal mortality and morbidity were low after following a management strategy based on duration of tachycardia, degree of heart failure, and biophysical profile combined with vigilant ongoing fetal surveillance. PMID- 11004363 TI - Downregulation of a mitogen-activated protein kinase signaling pathway in the placentas of women with preeclampsia. AB - OBJECTIVE: To investigate whether the activity of the three mitogen-activated protein kinases (Jun aminoterminal kinase, extracellular regulated kinase, and p38) is altered in placental tissue of women with preeclampsia and hemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. METHODS: Placental activity (measured by immunoprecipitation-kinase assay) and protein expression (measured by western blot) of Jun aminoterminal kinase, extracellular regulated kinase, and p38 mitogen-activated protein kinase were measured in four groups of eight women each with preeclampsia, HELLP syndrome, and normal vaginal or cesarean deliveries. To further characterize the Jun aminoterminal kinase signal transduction pathway, phosphorylation of c-Jun, a downstream effector of Jun aminoterminal kinase- mitogen-activated protein kinase, was analyzed by western blotting, and the activity of Rac1, an upstream activator of the Jun aminoterminal kinase signaling pathway, was determined by pull-down assay. RESULTS: The activity of Jun aminoterminal kinase was significantly lower in placentas of women with preeclampsia or HELLP syndrome compared with those who had normal vaginal or cesarean delivery, whereas levels of Jun aminoterminal kinase protein expression were similar. Phosphorylation of the transcription factor c-Jun and Rac1 activity also were significantly lower in women with preeclampsia and HELLP than in controls. p38 mitogen-activated protein kinase activity was significantly higher in women with preeclampsia than with HELLP syndrome. There was no change in extracellular regulated kinase activity or protein expression between subgroups. CONCLUSION: In placentas of women with preeclampsia or HELLP syndrome, a Rac1-Jun aminoterminal kinase-c-Jun-dependent signal transduction pathway was downregulated. PMID- 11004365 TI - Accuracy of the pelvic examination in detecting adnexal masses. AB - OBJECTIVE: To estimate sensitivity, specificity, positive predictive value, Youden J statistic, and likelihood ratio of pelvic examination in adnexal assessment under ideal circumstances. METHODS: One hundred forty women consented to have pelvic examinations under general anesthesia before laparoscopy or laparotomy. They were assigned to examiners masked to indications for surgery, including attending gynecologists, gynecology residents, and medical students. Surgeons' findings were compared with examiners' findings. Variables assessed included adnexal diameter, presence of adnexal masses, and effect modifiers such as examiner experience and body mass index. RESULTS: Forty-nine left adnexal and 33 right adnexal masses were found during surgery. Examiners tended to underestimate adnexal size. Sensitivity of pelvic examinations for detecting left adnexal masses was 0.23-0.36 and for right adnexal masses was 0.15-0.28. Positive predictive value was low for left adnexal masses (0.50-0. 69) and right adnexal masses (0.26-0.39). Differences among examiner groups were not statistically significant. Patient obesity noticeably reduced detection of adnexal masses on either side. CONCLUSION: Bimanual pelvic examination has marked limitations for evaluating adnexa, even with ideal circumstances. Experience during postgraduate training in gynecology did not seem to improve examination accuracy. Patient characteristics such as obesity, uterine size, and abdominal scars limit the accurate palpation of the adnexa. PMID- 11004364 TI - Alpha-fetoprotein and hematopoietic growth factors in amniotic fluid. AB - OBJECTIVE: To determine whether a relationship exists between alpha-fetoprotein (AFP) and hematopoietic growth factors in amniotic fluid. METHODS: Forty-one women at 15 weeks' gestation were included in the study. Gestational age was assessed by obtaining a reliable menstrual history and scanning. Amniocentesis was performed, and each woman subsequently delivered anatomically and chromosomally normal infants. The level of AFP was determined using a standard automated procedure. The concentrations of stem cell factor, interleukin 3, interleukin 6, erythropoietin, and granulocyte colony-stimulating factor (G-CSF) were measured using a commercially available immunoassay. The relationships between AFP and the studied cytokines were evaluated using the Pearson linear correlation test. Significant correlations were studied further by linear and nonlinear regression to obtain the best predictive model. RESULTS: There was a significant correlation between AFP and stem cell factor (r =.47, P =.002). No significant correlations between AFP and the rest of the studied cytokines were found (r = -.07, r =.02, r = -.02, and r = -.11 for erythropoietin, G-CSF, interleukin 3, and interleukin 6, respectively). CONCLUSION: Alpha-fetoprotein is significantly correlated with stem cell factor in early pregnancy and might play a role in fetal hematopoiesis. PMID- 11004366 TI - Clinical presentation of enterocele. AB - OBJECTIVE: To characterize preoperative signs and symptoms in women with and without enteroceles. METHODS: Three hundred ten women completed preoperative questionnaires and had prolapses graded according to the International Continence Society system. Signs and symptoms in 77 women (25%) with enteroceles confirmed at surgery were compared with those in 233 women without enteroceles. Comparisons were tested for statistical significance with chi(2) tests, Fisher exact tests, Wilcoxon rank-sum tests, and analysis of covariance. RESULTS: Women with enteroceles were statistically significantly older (median 67 versus 59 years, P <.001) and more likely to be postmenopausal (88% versus 76%, P =.04). More women with enteroceles had histories of hysterectomies (76% versus 39%, P =.001) and vaginal prolapse repairs (24% versus 11%, P =.008). Women with enteroceles had more advanced prolapses at points Ap, Bp, and C (all P <.001) but not point D. There were no significant differences in symptoms related to bowel function (infrequent bowel movements, straining, manual evacuation, and fecal incontinence) in women with and without enteroceles. Women with enteroceles were more bothered by symptoms caused by vaginal prolapse than women without enteroceles, but not after we controlled for stage of prolapse. CONCLUSION: Women with enteroceles have more advanced apical and posterior vaginal prolapses than women without enteroceles, but do not differ from them in bowel function. PMID- 11004367 TI - Early feeding and the incidence of gastrointestinal symptoms after major gynecologic surgery. AB - OBJECTIVE: To compare early feeding with traditional postoperative dietary management for development of postoperative gastrointestinal symptoms, including ileus after major gynecologic surgery for benign conditions. METHODS: Women who had major gynecologic surgery for benign conditions were randomly allocated to early feeding of low residue diets 6 hours postoperatively or traditional dietary management of clear liquids with normal bowel sounds, and regular diet with passage of flatus. Demographic and perioperative data were collected, and patients answered questionnaires on their perception of bowel function and pain using the McGill Pain Scale. Power analysis found that 130 women were needed to find a twofold greater incidence of ileus in the early feeding group with 80% power and alpha =.05. RESULTS: Complete data were available for 139 women, 67 allocated to the early feeding group and 72 to the late feeding group. The incidence of postoperative ileus for the study population was 4.4% and did not differ between groups (early 3% versus late 5. 8%, P =.68). There were no differences in patient demographics, surgical procedures, anesthesia used, and intraoperative complications between groups. With the exception of more complaints of nausea in the late feeding group (23% versus 13%, P =.04), there were no differences in other postoperative variables, including other perioperative complications, pain medicine requirements, fluid and caloric intake, median pain scores, and gastrointestinal function. The low incidence of perioperative complications made the power to detect differences between groups low. CONCLUSION: Low residue diet 6 hours after major gynecologic surgery for benign indications was not associated with increased postoperative gastrointestinal complaints, including ileus. PMID- 11004368 TI - Physical activity and reduced risk of ovarian cancer. AB - OBJECTIVE: To examine the relation between leisure-time physical activity and ovarian cancer. METHODS: We used data from a population based case-control study. Cases (n = 767) were women 20-69 years of age in whom epithelial ovarian cancer was diagnosed during 1994-1998 and who resided in a defined region of Pennsylvania, New Jersey, or Delaware. Controls (n = 1367) were ascertained by using random-digit dialing and Health Care Financing Administration files and were frequency-matched to cases for age and county of residence. Information on lifetime leisure-time physical activity was obtained during in-person interviews. RESULTS: Leisure-time physical activity was significantly associated with reduced occurrence of ovarian cancer (P =.01). After adjustment for age, parity, oral contraceptive use, tubal ligation, family history of ovarian cancer, race, and body mass index, women with the highest level of activity had an odds ratio of 0.73 (95% CI 0.56, 0.94) for ovarian cancer compared with women with the lowest level of activity. When the relation was analyzed by various recalled time periods during life, the odds ratios for the highest versus the lowest category of activity at ages 14-17, 18-21, 22-29, 30-39, 40-49, and >50 years ranged from 0.64-0.78. CONCLUSION: Leisure-time physical activity is associated with reduced occurrence of epithelial ovarian cancer. PMID- 11004369 TI - Angiogenesis of endometrial carcinomas assessed by measurement of intratumoral blood flow, microvessel density, and vascular endothelial growth factor levels. AB - OBJECTIVE: To evaluate the relationship between blood flow in the tumor assessed by color Doppler ultrasound, microvessel density, and vascular endothelial growth factor levels in endometrial carcinoma. METHODS: Forty-nine patients undergoing surgery for endometrial carcinoma were enrolled. Transvaginal color Doppler ultrasound was performed preoperatively and the lowest resistance index (RI) in the tumor was recorded for analysis. Vascular endothelial growth factor in the tumor was quantified by enzyme immunoassay. The microvessel density of the excised tumor was assessed immunohistochemically. The relationships between the corresponding RI, microvessel density, and vascular endothelial growth factor level of the tumor tissues and clinical and pathologic parameters were analyzed. RESULTS: Significantly lower RIs were noted in tumors of stage II or greater (0.37 compared with 0.50, P <.001), of high histologic grade (grade 3) (0.34 compared with 0.49, P =.004), with deep myometrial invasion (one-half depth or greater) (0.39 compared with 0.49, P =.002), with lymphovascular emboli (0.38 compared with 0.49, P <.001), or with lymph node metastasis (0.30 compared with 0.49, P <.001) compared with stage I tumors and tumors of histologic grade 1 or 2, with superficial myometrial invasion, without lymphovascular emboli, or with no lymph node metastasis. Increased vascular endothelial growth factor levels and microvessel density (x200 field) also were detected in tumors of stage II or greater (975 compared with 129 pg/mg, P =.014; and 88 compared with 61, P =.018, respectively), with lymphovascular emboli (1138 compared with 120 pg/mg, P =.002; and 86 compared with 63, P =.023), or with lymph node metastasis (1011 compared with 95 pg/mg, P <.001; and 98 compared with 61, P =. 019). Resistance index, microvessel density, and vascular endothelial growth factor levels in the tumor showed linear correlations (RI compared with microvessel density: r = -.32, P =. 03; RI compared with vascular endothelial growth factor levels: r = -.40, P =.004; microvessel density compared with vascular endothelial growth factor levels: r =.36, P =.011). CONCLUSION: Blood flow assessed by color Doppler ultrasound has histologic and biologic correlations with angiogenesis and vascular endothelial growth factor levels and might play an important role in predicting tumor progression and metastasis in endometrial carcinoma. PMID- 11004370 TI - Differences in perineal lacerations in black and white primiparas. AB - OBJECTIVE: To test the null hypothesis that there are no differences in incidence of perineal and vaginal lacerations in primiparous black and white women. METHODS: We reviewed University of Michigan Hospital delivery records, from July 1996 to December 1998, of black and white women 18 years and older and at least 35 weeks' gestation who had their first vaginal delivery. Birth weight, episiotomy, gestational age, laceration, length of second stage, oxytocin use, epidural use, and operative vaginal delivery were analyzed by univariable and multivariable tests. RESULTS: We analyzed 176 black women (mean age +/- standard deviation 23.7 +/- 4.7 years; range 18-41 years) and 1633 white women (27.8 +/- 5.4 years; 18-49 years; P <.001). Black women were less likely to have second, third, or fourth degree lacerations (43% compared with 59%; P <.001). The mean length of second stage of labor was shorter in the black women (73 +/- 69 minutes; range 3-494 minutes compared with 106 +/- 78 minutes; range 2-642 minutes; P <.001). Infants of black women weighed less (3292 +/- 490 g; 1990-5190 g compared with 3429 +/- 470 g; 1860-4950 g; P <.001). Multivariable analysis showed that black women were twice as likely to deliver with intact perineums than white women (P <.001). CONCLUSION: Black primiparas were less likely to deliver with second-degree or greater lacerations and more likely to deliver with their perineums intact. PMID- 11004372 TI - Do we need a subspecialty of female pelvic medicine and reconstructive surgery? PMID- 11004371 TI - Validity of adolescent and young adult self-report of Papanicolaou smear results. AB - OBJECTIVE: To assess the validity of adolescent and young adult report of Papanicolaou smear results and to determine sociodemographic, cognitive, and behavioral factors associated with incorrect reporting. METHODS: We conducted a cross-sectional study of 477 female subjects aged 12 to 24 years who attended an adolescent clinic and had a previous Papanicolaou smear. Subjects completed a self-administered survey assessing self-report of Papanicolaou smear results, knowledge about Papanicolaou smears and human papillomavirus (HPV), attitudes about Papanicolaou screening and follow-up, and risk behaviors. The sensitivity, specificity, positive predictive value, and negative predictive value of self reported results were calculated using the cytology report as the standard. Variables significantly associated with incorrect reporting were entered into logistic regression models controlling for age and race to determine independent predictors for incorrect reporting. RESULTS: Of the 477 participants, 128 (27%) had abnormal cytology reports and 66 (14%) had incorrect self-reports. Sensitivity of self-report was 0.79, specificity 0.89, positive predictive value 0.72, negative predictive value 0.92, and kappa (kappa) 0.66. The adjusted odds ratios (OR) and 95% confidence intervals (CI) of the variables comprising a logistic regression model predicting incorrect reporting were an HPV knowledge source of zero (OR 2.4, CI 1.0, 5.8), low perceived communication with the provider (OR 2.1, CI 1.1, 4.0), and no contraception at last intercourse (OR 5.5, CI 2.7, 11.0). CONCLUSION: The validity of adolescent and young adult self reported Papanicolaou smear result is high, except among those who lack knowledge of HPV, perceive poor communication with the provider, and use contraception inconsistently. PMID- 11004373 TI - Laparoscopic secondary port conversion using a reusable blunt conical trocar. AB - BACKGROUND: During operative laparoscopy, large (10 mm or more) ancillary ports are often used for instrumentation and tissue removal. Although sharp pyramidal trocars can be used to place these ports, their use appears to increase the risk of vessel injury and herniation. We describe a simple and cost-effective technique for converting a 5-mm port to a 10- or 12-mm port using a blunt conical trocar. TECHNIQUE: When a larger port is required, a previously placed 5-mm port is removed, and the skin incision is lengthened. A reusable 10- or 12-mm blunt conical trocar with a threaded sleeve is placed through the incision. The fascial defect is located by probing and is dilated gently with the blunt tip. Once the tip is through the fascia, it is advanced through the peritoneal defect with a clockwise, twisting motion. Afterwards, the fascial defect is closed with a single, interrupted absorbable suture. EXPERIENCE: We have had no complications or difficulty when using this technique in 26 cases, either during or after surgery. CONCLUSION: A reusable blunt conical trocar is a simple, safe, and cost effective instrument for converting a 5-mm laparoscopic port into a 10- or 12-mm port. PMID- 11004375 TI - In this issue PMID- 11004374 TI - Amniotic fluid volume estimation and the biophysical profile: a confusion of criteria. AB - The biophysical profile (BPP) can be used as an initial test of fetal health and as a secondary back-up assessment of fetuses at risk of adverse outcomes when preliminary evaluations are not reassuring. The BPP evaluates five characteristics: fetal movement, tone, breathing, heart reactivity, and amniotic fluid (AF) volume estimation. Three of the most frequently used obstetric textbooks define adequate AF volume differently. In two of the three, the stated method of evaluating AF volume differs from that actually used by the referenced authors. We reviewed articles by Manning and found that his methodology changed from a 1-cm pocket in one plane to a 1-cm pocket in two perpendicular planes, and finally to a 2-cm vertical pocket with a 1-cm horizontal measurement. The 2 x 2 cm pocket is a fourth methodology that has been introduced recently. It is not known how often and in which groups each of the four methods has been used to evaluate abnormal AF volumes. The relevance and importance of determining precisely the ultrasound measurement actually used for investigations are emphasized by looking at women with AF indices < or = 5. Fifty-three percent of those women had a 2 x 2 pocket, 72% had a 2 x 1 pocket, and 95% had a 1 x 1 pocket. The diagnosis of low fluid can lead to additional testing, hydration, and intervention, so the importance of a universal definition linked with pregnancy outcomes cannot be overemphasized. PMID- 11004376 TI - Paracelsus and mechanical ventilation. PMID- 11004377 TI - Automatic and manual mechanical external chest compression devices for cardiopulmonary resuscitation. PMID- 11004378 TI - Chronicity of memory impairment in long-term out-of-hospital cardiac arrest survivors. AB - As a result of out-of-hospital defibrillation initiatives, many cities have an increasing population of out-of-hospital cardiac arrest survivors. We previously identified that one third of these patients suffer memory problems in the months after resuscitation. The pattern of memory impairment (impaired recall memory and intact recognition memory) is suggestive of hippocampal damage. In this study we followed up ten subjects who had previously been found to have memory impairment after their cardiac arrest. To assess the chronicity of this memory impairment, we re-tested memory function approximately 3 years after the index events. These subjects were compared with age and sex matched control subjects with previous myocardial infarction and no cardiac arrest. Memory was assessed using the Rivermead Behavioural Memory test (RBMT). To further assess recall and recognition memory we used the Doors and People test (DPT), which is specifically designed to identify deficits in these functions. RBMT scores declined significantly in both groups compared with the original assessment 8 months after cardiac arrest, possibly an effect of ageing-control group: mean (S.D.) 22.2 (1. 4)-18.4 (2.9); cardiac arrest group: 16.1 (2.7)-14.6 (4.4). The inter-group difference in RBMT score remained significant (P=0.001). DPT scores were poor in the cardiac arrest group (mean (S.D.) total 5.8 (2.8)), compared with the control group (10.8 (3.4)) who scored normally. Both recall and recognition memory were poor in the cardiac arrest group. We conclude that the memory deficits that we previously observed in cardiac arrest victims are persistent. Both recall and recognition memory are affected, implying that non-selective brain injury may be the mechanism. PMID- 11004379 TI - Vasopressin pressor effects in critically ill children during evaluation for brain death and organ recovery. AB - BACKGROUND: Vasopressin (VP) shows promise as a pressor agent in animals and adult human cardiac arrest and resuscitation, but has not been studied for pressor effect in critically ill or arrested children. VP infusion is routine treatment for diabetes insipidus during brain death evaluation and organ recovery. We hypothesized that low dose VP infusion during organ recovery in critically ill children exerts a pressor effect, without major organ toxicity. METHODS: 34 VP-treated and 29 age-matched critically ill controls (C) < or =18 years were retrospectively reviewed during brain death evaluation and organ recovery. VP infusion protocol titrated VP dose clinically to urine output, with high variability. Pressor and inotrope management was titrated clinically to BP, cerebral perfusion and central venous pressures (when available) and peripheral perfusion with similar protocol targets for pre-load in VP and C groups. Outcome measures include dose, type and number of pressors and inotropes. Organ function was assessed at recovery and 48 h post-transplant by independent surgeon and transplant program organ function criteria. Analysis by Odds Ratio (OR), and chi square. RESULTS: VP dose averaged 0.041+/-0.069 U/kg/h. Average baseline mean arterial pressure (MAP) before VP infusion was 79+/-17 mmHg VP and 76+/-14 mm Hg C (P=0.6). Subsequent average MAP were: 82+/-21 mmHgVP after VP infusion versus 71+/-16 mmHg C (P=0.01) and 80+/-14 mmHg VP versus 68+/-22 mmHg C (P=0.01). Ability to wean/stop pressors and inotropes was: dopamine (14/23) 42% VP versus (10/26) 38% C (P=0.75), dobutamine (4/7) 57% VP versus (0/6) 0% C (P=0.026), epinephrine (4/5) 80% VP versus (0/6) 0% C (P=0.006), norepinephrine/phenylephrine (4/4) 100% VP versus (2/5) 40% C (P=0. 057). Alpha agonist pressor dependence was successfully weaned from 7/9 (78%) VP versus 0/9 (0%) C: odds ratio=7.3, (P<0.01). There was no VP induced dysrhythmia, hypertension, anuria or toxicity reported. Good organ recovery function was not significantly different at recovery or 48 h post-transplant for kidney (79% VP versus 69% C, P=0.068), liver (87% VP versus 95% C, P=0.533), or heart (90% VP versus 71% C, P=0.11). CONCLUSIONS: Low dose vasopressin infusion exerts a pressor effect in critically ill children treated for diabetes insipidus during brain death and organ recovery. VP treated patients were 7.3 times more likely to wean from alpha agonists than comparably managed age matched controls, without adverse affect on transplant organ function. We speculate that further prospective assessment of VP safety and efficacy as a pressor adjunct for resuscitation of critically ill children is warranted. PMID- 11004380 TI - Excellent coronary perfusion pressure during cardiopulmonary resuscitation is not good enough to ensure long-term survival with good neurologic outcome: a porcine case report. AB - PURPOSE: To report a case of cerebral ischemia confirmed by magnetic resonance imaging after successful cardiopulmonary resuscitation (CPR) complicated by acute respiratory injury. MATERIALS AND METHODS: After 4 min of cardiac arrest, followed by 3 min of basic life support CPR, a female pig weighing 38 kg received every 5 min vasopressin (0.4, 0.4 and 0.8 U/kg). After 22 min of cardiac arrest, including 18 min of CPR, one defibrillation attempt employing 100 J resulted in return of spontaneous circulation. Neurological evaluation was performed 24 and 96 h after successful CPR. Magnetic resonance imaging was carried out 4 days after CPR using a clinical 1.5 T scanner. The magnetic resonance imaging protocol consisted of fast spinecho T2-weighted, as well as spinecho T1-weighted imaging of the brain. RESULTS: CPR with vasopressin resulted in excellent coronary perfusion pressure ranging between 35 and 60 mm Hg throughout CPR. Eight minutes after initiation of chest compressions, bleeding out of the tracheal tube occurred. This was later confirmed as originating from bilateral bloody pulmonary infiltrations, resulting in acute respiratory injury in the post-resuscitation phase. Ninety-six hours after successful CPR, magnetic resonance imaging revealed bilateral diffuse cerebral vasogenic edema. CONCLUSION: Although excellent coronary perfusion pressure renders a return of spontaneous circulation more likely, complications such as acute respiratory injury in the post-resuscitation phase have to be managed carefully in order to ensure good neurological recovery from cardiac arrest. PMID- 11004381 TI - Transthoracic monophasic and biphasic defibrillation in a swine model: a comparison of efficacy, ST segment changes, and postshock hemodynamics. AB - OBJECTIVE: Biphasic waveforms for transthoracic defibrillation (DF) have been tested extensively after brief (15 s) episodes of VF in animal models and in patients undergoing electrophysiologic testing. The purpose of this study was to compare the effects mono- and biphasic waveforms for DF on postdefibrillation ST segments and left ventricular pressure, markers of myocardial injury, after more extended periods of VF (30 and 90 s). METHODS: 21 anesthetized and instrumented swine were randomized to truncated exponential monophasic or biphasic waveform DF. VF was induced electrically and 30 s later, DF with the designated waveform was attempted with a shock dose of 200 J. If unsuccessful, 300 J and then 360 J were administered if necessary. Following return to control hemodynamic values and normalization of the surface ECG, VF was again induced and, after 90 s, DF was attempted as in the 30 s VF period. CPR was not performed during VF and each animal was countershocked with only one waveform for both VF episodes. Waveforms were compared for frequency of first shock defibrillation success, surface ECG indicators of myocardial injury (ST segment changes at 10, 20, and 30 s after countershock) and time to return to pre-VF hemodynamics after successful DF, an indicator of postshock ventricular function. RESULTS: Successful first shock conversion rates at 30 and 90 s were 60 and 63% for monophasic and 64 and 82% for biphasic (NS). Biphasic DF after 30 s produced ST segment changes (measured 10 s after DF) in 1/10 animals while six of eight animals in the monophasic group showed ST segment changes (P=0.013). After 90 s of VF, ST segment changes were observed in 6/8 in the monophasic group and 2/10 in the biphasic group (P=0.054). Differences in the time to hemodynamic recovery (return to control peak left ventricular pressure) were not observed between biphasic and monophasic waveforms after 30 or 90 s of VF. CONCLUSIONS: Monophasic and biphasic transthoracic defibrillation are equally effective in terminating VF of 30 and 90 s duration and restoring a perfusing rhythm. The biphasic waveform produced less ECG evidence of transient myocardial injury. However, there was no difference in the rate of return to control hemodynamics. ST segment changes following countershock of VF of brief duration are transient and of questionable significance. PMID- 11004382 TI - Effect of bystander cardiopulmonary resuscitation in out-of-hospital cardiac arrest patients in Sweden. AB - BACKGROUND: Information from the Swedish Cardiac Arrest Registry was used to investigate: (a) The proportion of patients suffering an out-of-hospital cardiac arrest who were given bystander cardiopulmonary resuscitation (B-CPR). (b) Where and by whom B-CPR was given. (c) The effect of B-CPR on survival. METHOD: a prospective, observational study of cardiac arrests reported to the Swedish Cardiac Arrest Registry. Analyses were based on standardised reports of out-of hospital cardiac arrests from ambulance organisations in Sweden, serving 60% of the Swedish population. From 1983 to 1995 approximately 15-20% of the population had been trained in CPR. RESULTS: Of 9877 patients, collected between January 1990 and May 1995, B-CPR was attempted in 36%. In 56% of these cases, the bystanders were lay persons and in 25% they were medical personnel. Most of the arrests took place at home (69%) and only 23% of these patients were given B-CPR in contrast to cardiac arrest in other places where 53% were given CPR. Survival to 1 month was significantly higher in all cases that received B-CPR (8.2 vs. 2.5%). The odds ratio for survival to 1 month with B-CPR was in a logistic regression analysis 2.5 (95% CI 1.9-3.1). CONCLUSIONS: In Sweden, the willingness and ability to perform B-CPR appears to be relatively widespread. More than half of B-CPR was performed by laypersons. B-CPR resulted in a two to threefold increase in survival. PMID- 11004383 TI - How bystanders perceive their cardiopulmonary resuscitation intervention; a qualitative study. AB - The importance of bystander cardiopulmonary resuscitation (CPR) prior to arrival of the emergency medical service is well documented. In Sweden, CPR is initiated prior to emergency medical services (EMS) arrival in about 30% of cardiac arrests out-of-hospital, a figure which should be improved urgently. To do so, it is of interest to know more about the bystanders' perceptions of their intervention. A qualitative method inspired by the phenomenographic approach was applied to 19 bystanders who had performed CPR. In the analysis, five main categories and 14 subcategories emerged. The main categories were: to have a sense of humanity, to have competence, to feel an obligation, to have courage and to feel exposed. Interviews described how humanity and concern for another human being were the foundation of their intervention. CPR training offers the possibility to give appropriate help in this emergency. If the aim of CPR training was extended beyond teaching the skill of CPR to include preparation of the rescuer for the intervention and his/her reactions, this might increase the number of people able to take action in the cardiac arrest situation. PMID- 11004384 TI - In-hospital resuscitation: association between ACLS training and survival to discharge. AB - CONTEXT: No data have been published on the relationship between advanced cardiac life support (ACLS) training of the individual who initiates resuscitation efforts and survival to discharge. OBJECTIVE: To determine whether patients whose arrests were discovered by nurses trained in ACLS had survival rates different from those discovered by nurses not trained in ACLS. DESIGN: Cohort case comparison. SETTING: A 550-bed, tertiary care center in central Georgia. SUBJECTS: Patients whose cardiopulmonary arrest was discovered by a nurse who activated the in-hospital resuscitation mechanism. MAIN OUTCOME MEASURE: Patient survival to discharge. RESULTS: Initial rhythm was strongly related to survival to discharge and individually associated with 57% of the variability in survival. Nurse's training in advanced cardiac life support was also strongly related to survival and individually associated with 29% of the variability. Combining both the variables determined 62% of the variability in survival to discharge. Patients discovered by an ACLS-trained nurse (n=88) were about four times more likely to survive (33 survivors, 38%) than were patients, discovered by a nurse without training in ACLS (n=29, three survivors, 10%). CONCLUSION: Arrest discovery by nurses trained in ACLS is significantly and dramatically associated with higher survival-to-discharge rates. PMID- 11004385 TI - Endobronchial application of high dose epinephrine in out of hospital cardiopulmonary resuscitation. PMID- 11004386 TI - A simple model of the thermal prebiotic oligomerization of amino acids. AB - We construct a probabilistic model with the aid of the Markov chain formalism to describe and give a physico-chemical justification to an oligomerization process of a set of amino acids under certain prebiotic conditions. Such chemical process shows a remarkable bias in the polymer products that our model can explain. Some predictions and limitations are also discussed. PMID- 11004387 TI - Complex calcium oscillations and the role of mitochondria and cytosolic proteins. AB - Intracellular calcium oscillations, which are oscillatory changes of cytosolic calcium concentration in response to agonist stimulation, are experimentally well observed in various living cells. Simple calcium oscillations represent the most common pattern and many mathematical models have been published to describe this type of oscillation. On the other hand, relatively few theoretical studies have been proposed to give an explanation of complex intracellular calcium oscillations, such as bursting and chaos. In this paper, we develop a new possible mechanism for complex calcium oscillations based on the interplay between three calcium stores in the cell: the endoplasmic reticulum (ER), mitochondria and cytosolic proteins. The majority ( approximately 80%) of calcium released from the ER is first very quickly sequestered by mitochondria. Afterwards, a much slower release of calcium from the mitochondria serves as the calcium supply for the intermediate calcium exchanges between the ER and the cytosolic proteins causing bursting calcium oscillations. Depending on the permeability of the ER channels and on the kinetic properties of calcium binding to the cytosolic proteins, different patterns of complex calcium oscillations appear. With our model, we are able to explain simple calcium oscillations, bursting and chaos. Chaos is also observed for calcium oscillations in the bursting mode. PMID- 11004388 TI - Computing with DNA by operating on plasmids. AB - A new method of computing using DNA plasmids is introduced and the potential advantages are listed. The new method is illustrated by reporting a laboratory computation of an instance of the NP-complete algorithmic problem of computing the cardinal number of a maximal independent subset of the vertex set of a graph. A circular DNA plasmid, specifically designed for this method of molecular computing, was constructed. This computational plasmid contains a specially inserted series of DNA sequence segments, each of which is bordered by a characteristic pair of restriction enzyme sites. For the computation reported here, the DNA sequence segments of this series were used to represent the vertices of the graph being investigated. By applying a scheme of enzymatic treatments to the computational plasmids, modified plasmids were generated from which the solution of the computational problem was selected. This new method of computing is applicable to a wide variety of algorithmic problems. Further computations in this style are in progress. PMID- 11004389 TI - Canalization as a non-genetic source of adaptiveness during morphogenesis: experimental evidence from analysis of reproductive development in Sorghum bicolor. AB - In Sorghum bicolor, perturbations in reproductive development observed following salt-treatment also influence progeny grown in the absence of NaCl. However, a developmental reversion of these modifications may be observed throughout two successive generations. This response, termed canalization, does not spontaneously occur following growth in the absence of NaCl, but is triggered by the level of perturbation in parental expression of reproductive characters. Moreover, canalization is not specific to the perturbed character, but it includes modifications in reproductive development as a whole. A decrease in developmental variability coincides with amplitude of the developmental reversion. This phenomenon is interpreted as an evidence for orientation of the developmental process towards the lowest free-energy state of the 'epigenetic landscape'. Involvement of this phenomenon of canalization in developmental stability, adaptiveness, and evolution is discussed. Moreover, these results point to the need for a posteriori methods of investigations in order to analyze self-organized transformations in biological systems. PMID- 11004390 TI - Probing quantum coherence in a biological system by means of DNA amplification. AB - As a result of rapid decoherence, quantum effects in biological systems are usually confined to single electron or hydrogen delocalizations. In principle, molecular interactions at high temperatures can be guided by quantum coherence if embedded in a dynamics preventing decoherence. This was experimentally investigated by analyzing the thermodynamics, kinetics, and quantum mechanics of the primer/template duplex formation during DNA amplification by polymerase chain reaction. The structures of the two oligonucleotide primers used for amplification of a cDNA template were derived either from a repetitive motif or a fractal distribution of nucleotide residues. Contrary to the computer-based calculation of the primer melting temperatures (T(m)) that predicted a higher T(m) for the non-fractal primer due to nearest-neighbor effects, it was found that the T(m) of the non-fractal primer was actually 2 degrees C lower than that of its fractal counterpart. A thermodynamic analysis of the amplification reaction indicated that the primer annealing process followed Bose-Einstein instead of Boltzmann statistics, with an additional binding potential of mu=500 J/mol or 10(-21) J/molecule due to a superposition of binding states within the primer/template duplex. The temporal evolution of the Bose-Einstein state was determined by enzyme kinetic analysis of the association of the primer/template duplex to Taq polymerase. Assuming that collision with the enzyme interrupted the superposition, it was found that the Bose-Einstein state lasted for t(dec)=0.7x10(-12) s, corresponding to the energy dispersion (DeltaE) of quantum coherent states (mu=DeltaE>/=h/t(dec)). A quantum mechanical analysis revealed that the coherent state was stabilized by almost vanishing separation energies between distinct binding states during a temperature-driven shifting of the two DNA strands in the primer/template duplex. The additional binding potential is suggested to arise from a short-lived electron tunneling as the result of overlapping orbitals along the axis of the primer/template duplex. This effect was unique to the fractal primer due to the number of binding states that remained almost constant, irrespective of the size of shifting. It is suggested that fractal structures found in proteins or other macromolecules may facilitate a short-lived quantum coherent superposition of binding states. This may stabilize molecular complexes for rapid sorting of correct-from-false binding, e.g. during folding or association of macromolecules. The experimental model described in this paper provides a low-cost tool for simulating and probing quantum coherence in a biological system. PMID- 11004391 TI - The iron dependent regulatory protein IdeR (DtxR) of Rhodococcus equi. AB - This paper reports the presence of an ideR gene, which encodes an iron-dependent regulatory protein, in Rhodococcus erythropolis and in the intracellular pathogen Rhodococcus equi. The ideR gene of the latter encoded a protein of 230 amino acids with a molecular mass of 25619. The alpha-helices forming the helix-turn helix motif of the R. equi protein were identical to those of the DtxR protein of Corynebacterium diphtheriae, which is an IdeR homologue. This indicates that the two proteins bind to the same DNA binding site. This was confirmed following expression of IdeR in Escherichia coli, which showed that the IdeR protein could repress transcription of the tox promoter of C. diphtheriae in an iron dependent manner. An open reading frame specifying a 283-amino acid polypeptide similar to galE encoding UDP-galactose 4-epimerase was present downstream of the ideR gene. PMID- 11004392 TI - Inducible stx2 phages are lysogenized in the enteroaggregative and other phenotypic Escherichia coli O86:HNM isolated from patients. AB - We characterized two Shiga toxin-producing Escherichia coli (STEC) O86:HNM isolates from a patient with hemolytic uremic syndrome (HUS) or bloody diarrhea. Both of them did not possess the eaeA gene. However, the isolate from a HUS patient carried genetic markers of enteroaggregative E. coli (EAEC) and showed aggregative adherence pattern to HEp-2 cells. The other isolate from bloody diarrhea, which was negative with EAEC markers, was diffusely adhered to HEp-2 cells. The stx2 gene in both E. coli O86:HNM strains was encoded in each infectious phage, which was partially homologous to that of strain EDL933, a STEC O157:H7. These results will help to explain the genotypic divergences of STEC. PMID- 11004393 TI - The colanic acid gene cluster of Salmonella enterica has a complex history. AB - The colanic acid gene cluster of Salmonella enterica LT2 was sequenced and compared with that of Escherichia coli K-12. The two clusters are similar with divergence slightly higher than average for genes of the two species. The cluster was divided into four blocks by GC content and seems likely to have transferred from a higher GC content species to the ancestor of E. coli and S. enterica. All 19 genes of K-12 and 13 genes of LT2 appear to have undergone random genetic drift with amelioration of the GC content. However, in the case of S. enterica, we believe that the six genes of the GDP-fucose pathway group were replaced relatively recently by genes closely related to those of the original donor species. Two repetitive elements were observed: a bacterial interspersed mosaic element in the intergenic region between wzx and wcaK in K-12 only and a RSA (repetitive sequence element) sequence between wcaJ and wzx in LT2 only. PMID- 11004394 TI - Molecular differentiation of Bifidobacterium species with amplified ribosomal DNA restriction analysis and alignment of short regions of the ldh gene. AB - The differentiation of Bifidobacterium species was performed with specific primers using the PCR technique, the amplified ribosomal DNA restriction analysis (ARDRA) technique based on reports on the sequence of the 16S rRNA gene and speciation based on a short region of the ldh gene. Four specific primer sets were developed for each of the Bifidobacterium species, B. animalis, B. infantis and B. longum. The use of the ARDRA method made it possible to discriminate between B. infantis, B. longum and B. animalis with the combination of BamHI, TaqI and Sau3AI restriction enzymes. The ldh gene sequences of 309-312 bp were determined for 19 Bifidobacterium strains. Alignment of these short regions of the ldh gene confirmed that it is possible to distinguish between B. longum and B. infantis but not between B. lactis and B. animalis. PMID- 11004395 TI - Effects of tubulin assembly inhibitors on cell division in prokaryotes in vivo. AB - The bacterial cell division protein FtsZ is a structural analogue of tubulin. Bacterial mutants in which the ftsZ gene is inactivated are unable to divide. Numerous inhibitors of tubulin assembly are known, some of which are used as fungicides. The strong structural homology between FtsZ and tubulin raises the possibility that some of these inhibitors could affect bacterial cell division. Here we report that the tubulin assembly inhibitors thiabendazole and 2 methylbenzimidazole cause cell elongation in Escherichia coli and cyanobacteria. PMID- 11004396 TI - The relationship between glycogen synthesis, biofilm formation and virulence in salmonella enteritidis. AB - Salmonella enteritidis accumulated large quantities of intracellular polysaccharide when grown in unrestricted nutrient conditions. Dense, abundant cytoplasmic granules were observed by electron microscopy in sections stained by the periodic acid-chlorite technique, indicating that the polysaccharide was of the glycogen type. When biofilm-producing S. enteritidis was pre-incubated in media containing increasing levels of glucose concentration, the levels of both cytoplasmic glycogen and biofilm rose correlatively to a point where a ceiling effect was observed. Studies carried out with activators and inhibitors of glycogen biosynthesis confirmed that biofilm was formed from glycogen cell stores. On the other hand, the virulence of the biofilm-producing strain in infected chickens increased proportionally to the amount of stored glycogen, suggesting a possible role of the glycogen depot in the virulence of S. enteritidis. PMID- 11004397 TI - The cloning of a new peroxidase found in lignocellulose cultures of Pleurotus eryngii and sequence comparison with other fungal peroxidases. AB - We report cloning and sequencing of gene ps1 encoding a versatile peroxidase combining catalytic properties of lignin peroxidase (LiP) and manganese peroxidase (MnP) isolated from lignocellulose cultures of the white-rot fungus Pleurotus eryngii. The gene contains 15 putative introns, and the deduced amino acid sequence consists of a 339-residue mature protein with a 31-residue signal peptide. Several putative response elements were identified in the promoter region. Amino acid residues involved in oxidation of Mn(2+) and aromatic substrates by direct electron transfer to heme and long-range electron transfer from superficial residues as predicted by analogy with Phanerochaete chrysosporium MnP and LiP, respectively. A dendrogram is presented illustrating sequence relationships between 29 fungal peroxidases. PMID- 11004398 TI - Peptide mimics elicit antibody responses against the outer-membrane lipooligosaccharide of group B neisseria meningitidis. AB - As an alternative approach towards the development of a meningococcal vaccine, the potential of peptide mimics of lipooligosaccharide (LOS) to elicit cross reactive immune responses against LOS was investigated. The heptapeptides SMYGSYN and APARQLP were identified by enrichment from a coliphage display library with a LOS-specific monoclonal antibody. Mice immunised with these peptides conjugated to diphtheria toxoid elicited a total IgG response to LOS with geometric mean titres 2-4 times higher compared with non-immunised controls. There was an increase in LOS-specific IgG1 immunoglobulin, whereas specific IgG2a and IgG3 decreased slightly in response to immunisation. The data demonstrated that peptide mimics can elicit immune responses against meningococcal LOS. PMID- 11004399 TI - Acetaldehyde metabolism by wine lactic acid bacteria. AB - Acetaldehyde is a volatile flavor compound present in many fermented foods and is important in the production of red and white wines. Nine strains of the genera Lactobacillus and Oenococcus were able to metabolize acetaldehyde in a resting cell system, whereas two Pediococcus strains were not. Acetic acid and ethanol were produced from its degradation. A Lactobacillus and an Oenococcus were able to degrade SO(2)-bound acetaldehyde, as well. A coincubation of resting cells of Saccharomyces bayanus Premiere Cuvee and Oenococcus oeni Lo111 showed that strain Lo111 metabolized acetaldehyde produced by the yeast. The ability of malolactic bacteria to degrade free and SO(2)-bound acetaldehyde has implications for sensory and color qualities and the use of SO(2) in wine. PMID- 11004400 TI - The immunosuppressive drug leflunomide affects mating-pheromone response and sporulation by different mechanisms in Saccharomyces cerevisiae. AB - Leflunomide (LFM) is a novel anti-inflammatory and immunosuppressive drug, and inhibits the growth of cytokine-stimulated lymphoid cells in vitro. The effect of LFM on haploid and diploid cells of Saccharomyces cerevisiae was investigated to elucidate the molecular mechanism of action of the drug. Using a halo assay, LFM was shown to enhance the cell cycle arrest of haploid cells induced by mating pheromone alpha-factor. LFM also inhibited sporulation of diploid cells completely. S. cerevisiae genes which were cloned to suppress the anti proliferative effect when present in increased copy number were introduced and examined for their activity to suppress the effect of LFM. Out of them, MLF4/SSH4, was found to suppress the sporulation-inhibitory effect of LFM. However, MLF4 failed to suppress the enhancing effect of LFM on pheromone response. Thus, LFM is suggested to act on haploid and diploid cells by different mechanisms. PMID- 11004401 TI - Elevated zinc induces siderophore biosynthesis genes and a zntA-like gene in Pseudomonas fluorescens. AB - Zinc-regulated genes were analyzed in Pseudomonas fluorescens employing mutagenesis with a reporter gene transposon. Six mutants responded with increased gene expression to elevated concentrations of zinc. Genetic and biochemical analysis revealed that in four of the six mutants the transposon had inserted into genes essential for the biosynthesis of the siderophore pyoverdine. The growth of one of the mutants was severely impaired in the presence of elevated concentrations of cadmium and zinc ions. In this mutant, the transposon had inserted in a gene with high similarity to P-type ATPases involved in zinc and cadmium ion transport. Four mutants reacted with reduced gene expression to elevated concentrations of zinc. One of these mutants was sensitive to zinc, cadmium and copper ions. The genetic region targeted in this mutant did not show similarity to any known gene. PMID- 11004402 TI - Survival of free DNA encoding antibiotic resistance from transgenic maize and the transformation activity of DNA in ovine saliva, ovine rumen fluid and silage effluent. AB - To assess the likelihood that the bla gene present in a transgenic maize line may transfer from plant material to the microflora associated with animal feeds, we have examined the survival of free DNA in maize silage effluent, ovine rumen fluid and ovine saliva. Plasmid DNA that had previously been exposed to freshly sampled ovine saliva was capable of transforming competent Escherichia coli cells to ampicillin resistance even after 24 h, implying that DNA released from the diet could provide a source of transforming DNA in the oral cavity of sheep. Although target DNA sequences could be amplified by polymerase chain reaction from plasmid DNA after a 30-min incubation in silage effluent and rumen contents, only short term biological activity, lasting less than 1 min, was observed in these environments, as shown by transformation to antibiotic resistance. These experiments were performed under in vitro conditions; therefore further studies are needed to elucidate the biological significance of free DNA in the rumen and oral cavities of sheep and in silage effluent. PMID- 11004403 TI - Purification, characterisation, cloning and sequencing of the gene encoding oligopeptidase PepO from Streptococcus thermophilus A. AB - The oligopeptidase PepO from Streptococcus thermophilus A was purified to protein homogeneity by a five-step chromatography procedure. It was estimated to be a serine metallopeptidase of 70 kDa, with maximal activity at pH 6.5 and 41 degrees C. PepO has endopeptidase activity on oligopeptides composed of between five and 30 amino acids. PepO was demonstrated to be active and stable at the pH, temperature and salt concentrations found in Swiss-type cheese during ripening. Using a battery of PCR techniques, the pepO gene was amplified, subcloned and sequenced, revealing an open reading frame of 1893 nucleotides. The amino acid sequence analysis of the pepO gene-translation product shows homology with PepO enzymes from other lactic acid bacteria and contains the signature sequence of the metallopeptidase family. PMID- 11004404 TI - Cloning of the sth gene from Azotobacter vinelandii and construction of chimeric soluble pyridine nucleotide transhydrogenases. AB - The gene encoding the soluble pyridine nucleotide transhydrogenase (STH) of Azotobacter vinelandii was cloned and sequenced. This is the third sth gene identified and further defines a new subfamily within the flavoprotein disulfide oxidoreductases. The three STHs identified all lack one of the redox active cysteines that are characteristic for this large family of enzymes, and instead they contain a conserved threonine residue at this position. The recombinant A. vinelandii enzyme was purified to homogeneity and shown to form filamentous structures different from those of Pseudomonas fluorescens and Escherichia coli STH. Chimeric STHs were constructed which showed that the C-terminal region is important for polymer formation. The A. vinelandii STH containing the C-terminal region of P. fluorescens or E. coli STH showed structures resembling those of the STH contributing the C-terminal portion of the protein. PMID- 11004405 TI - A novel technique for monitoring the development of bacterial biofilms in human periodontal pockets. AB - A new technique is presented for analyzing subgingival bacterial plaque. Different materials (polytetrafluoroethylene, gold, dentin) kept for several days in periodontal pockets of patients suffering from periodontitis were analyzed by electron microscopy and fluorescence in situ hybridization (FISH). Those parts of the carriers extending into the deepest zone of the pockets were predominantly colonized by spirochetes and Gram-negative bacteria whereas those segments in contact with a shallower region were colonized by streptococci. Independent of the material used, the bacterial colonization of the carriers appears to be similar. FISH using eubacteria- and species-specific oligonucleotides on semi thin cross-sections of the carriers in combination with confocal laser scanning microscopy allowed detailed analysis of the architecture of biofilms and identification of putative periodontal pathogens with single cell resolution. PMID- 11004406 TI - The alternative sigma factor sigma(28) of the extreme thermophile Aquifex aeolicus restores motility to an Escherichia coli fliA mutant. AB - Sigma factor sigma(28) (sigma(F), FliA, SigD) directs RNA polymerase to transcribe the genes required for flagellar biosynthesis and chemotaxis in many bacteria, including Bacillus subtilis, Legionella pneumophila, Salmonella typhimurium, Escherichia coli, Yersinia enterolytica, Treponema maltophilum and Pseudomonas aeruginosa. Remarkably the fliA gene from the extreme thermophile Aquifex aeolicus restored motility to the E. coli mutant at relatively low temperature, albeit partially. This clearly demonstrates that A. aeolicus sigma(28) is able to direct RNA polymerase to E. coli sigma(28)-dependent promoters and take part in the complex interactions required to support transcription of the flagellar apparatus in vivo. The ability of A. aeolicus sigma(28) to function with mesophilic components shows that critical functional interactions made by these sigma factors are well conserved, and are not dependent upon high temperature. We over-produced and purified the sigma(28) protein and demonstrated binding to E. coli core RNA polymerase in vitro. In common with SigD from B. subtilis, but unlike most sigma factors, A. aeolicus sigma(28) showed DNA binding activity in vitro but there was no evidence of sequence specificity. We note that A. aeolicus sigma(28) is a good candidate for structural studies. PMID- 11004407 TI - Membrane vesicles released by Actinobacillus pleuropneumoniae contain proteases and Apx toxins. AB - Actinobacillus pleuropneumoniae serotype 1 releases vesicles containing proteases and Apx toxins into the culture medium. Vesicles were concentrated by ultracentrifugation and analyzed by electron microscopy and electrophoresis; their size ranged from 20 to 200 nm. A polyclonal antiserum raised against a purified high molecular mass secreted protease of serotype 1 recognized this protease on the surface of the vesicles by immunogold electron microscopy. Higher molecular mass polypeptides from vesicle extracts were recognized by the antiserum by Western immunoblot, indicating that the protease could form oligomers. However, these oligomers were not active against gelatin until secreted. Additionally, Apx toxins were also present in vesicles, and were recognized by Western immunoblot by an anti-serotype 1 toxins polyclonal serum. A. pleuropneumoniae antigens in vesicles were recognized by convalescent-phase pig sera from animals infected with serotype 1 or 5. The release of vesicles containing virulence factors could be a tissue damage mechanism in swine pleuropneumonia. PMID- 11004408 TI - Identification of novel metabolites in the degradation of phenanthrene by Sphingomonas sp. strain P2. AB - Sphingomonas sp. strain P2, which is capable of utilizing phenanthrene as a sole carbon and energy source, was isolated from petroleum-contaminated soil in Thailand. Gas chromatography-mass spectrometry and (1)H and (13)C nuclear magnetic resonance analyses revealed two novel metabolites from the phenanthrene degradation pathway. One was identified as 5,6-benzocoumarin, which was derived by dioxygenation at the 1- and 2-positions of phenanthrene, and the other was determined to be 1,5-dihydroxy-2-naphthoic acid. Other metabolites from phenanthrene degradation were identified as 7, 8-benzocoumarin, 1-hydroxy-2 naphthoic acid and coumarin. From these results, it is suggested that strain P2 can degrade phenanthrene via dioxygenation at both 1,2- and 3,4-positions followed by meta-cleavage. PMID- 11004409 TI - Identification of DNA amplifications near the center of the Streptomyces coelicolor M145 chromosome. AB - Linear streptomycete chromosomes frequently undergo spontaneous gross DNA rearrangements at the terminal regions. Large DNA deletions of the chromosome ends are in many cases associated with tandemly reiterated DNA amplifications, found at the border of the deletable areas. In contrast to previous reports, we have discovered amplifications near the center of the Streptomyces coelicolor M145 chromosome. The detected amplified units of DNA are 19.9 kb and 16 kb in length and exist in copy numbers of 30 and 40, respectively. Both amplifications were located in the same region and share at least 3.6 kb. PMID- 11004410 TI - Isolation of the dxr gene of Zymomonas mobilis and characterization of the 1 deoxy-D-xylulose 5-phosphate reductoisomerase. AB - The gene encoding the second enzyme of the 2C-methyl-D-erythritol 4-phosphate (MEP) pathway for isopentenyl diphosphate biosynthesis, 1-deoxy-D-xylulose 5 phosphate (DXP) reductoisomerase, was cloned and sequenced from Zymomonas mobilis. The deduced amino acid sequence showed the highest identity (48.2%) to the DXP reductoisomerase of Escherichia coli. Biochemical characterization of the purified DXP reductoisomerase showed a strict dependence of the enzyme on NADPH and divalent cations (Mn(2+), Co(2+) or Mg(2+)). The enzyme is a dimer with a molecular mass of 39 kDa per subunit and has a specific activity of 19.5 U mg protein(-1). Catalysis of the intramolecular rearrangement and reduction of DXP to MEP is competitively inhibited by the antibiotic fosmidomycin with a K(i) of 0.6 microM. PMID- 11004412 TI - Detection and characterisation of integrons in Salmonella enterica serotype enteritidis. AB - Integrons have been widely described among the Enterobacteriaceae including strains of multi-resistant Salmonella enterica serotype Typhimurium DT104; however, information with respect to the presence of integrons among S. enterica serotype Enteritidis strains is limited. Multi-resistant isolates of Enteritidis were screened for the presence of integrons using a PCR protocol. One integron was detected in all isolates that were resistant to sulfonamide and streptomycin. Characterisation of these isolates indicated an integron which ranged in size between 1000 and 2000 bp and which harboured a gene cassette encoding the ant(3") Ia gene specifying streptomycin and spectinomycin resistance. Further studies revealed the integrons to be located on large conjugative plasmids. This appears to be the first report of plasmid-borne integrons in Enteritidis. PMID- 11004411 TI - The predicted structure of photopexin from Photorhabdus shows the first haemopexin-like motif in prokaryotes. AB - The insect pathogenic bacterium Photorhabdus luminescens secretes several insecticidal high molecular mass 'toxin complexes'. Analysis of the putative pathogenicity island surrounding the toxin complex a (tca) locus revealed two open reading frames (ORFs) of unknown function. The predicted protein sequences of these ORFs show a repeated motif similar to those found in the vertebrate haem scavenging molecule haemopexin, limunectin (a phosphocholine binding protein from Limulus) and the C-terminal domains of matrix metalloproteinases (MMPs) (where they are thought to be important for cell attachment and adhesion). We have therefore named the operon photopexin AB and the putative encoded proteins 'photopexins' A and B (PpxA and PpxB). The predicted amino acid sequence of PpxA was modelled onto the crystal structure of a MMP. Our model predicts not only that PpxA and PpxB have beta-propeller domains but also that each haemopexin-like repeat corresponds to one blade of a propeller, suggesting the limunectin structure itself may also contain two or three such haemopexin-like propellers. The overall structure of PpxA has striking similarity to that of haemopexin suggesting that it may be used by the bacterium to scavenge iron containing compounds from insects. The implications for the potential role of Ppx proteins in pathogenicity are discussed. This is the first finding of a haemopexin-like repeat outside plants and animals. PMID- 11004413 TI - Phagocytosis, oxidative burst, and killing of Candida dubliniensis and Candida albicans by human neutrophils. AB - Candida dubliniensis is a phylogenetically closely related species to Candida albicans. So far virtually nothing is known about the virulence factors of C. dubliniensis. Cell surface hydrophobicity (CSH) plays a critical role in adhesion of microorganisms to phagocytic cells; hydrophobic cells of C. albicans have been reported to be less sensitive to phagocytic killing than hydrophilic cells. C. dubliniensis displays CSH at 37 degrees C in contrast to C. albicans. To elucidate this issue, we determined levels of phagocytosis, oxidative burst and killing by human neutrophils of C. dubliniensis (n=10) compared to C. albicans (n=10) both cultured at 37 degrees C. Obtained test results revealed no statistically significant differences between these two yeast species for the level of phagocytosis (77.3 vs. 76.2% after 60 min), evoked oxidative burst (64.5 vs. 67.3% after 30 min) and killing (72.7 vs. 73.1% after 240 min). Therefore, human neutrophils can be considered to be equally efficient against these two yeast species. PMID- 11004414 TI - Erratum to: "Stimulation of adhesiveness, infectivity, and competitiveness for nodulation of bradyrhizobium japonicum by its pretreatment with soybean seed lectin". PMID- 11004415 TI - Do potential patients prefer tissue plasminogen activator (TPA) over streptokinase (SK)? An evaluation of the risks and benefits of TPA from the patient's perspective. AB - BACKGROUND: In patients with acute myocardial infarction, TPA (compared to SK), has been shown to reduce the 30-day mortality rate at the expense of an increased rate of stroke. The assumption in the literature is that were it not for cost issues, all patients presenting with a myocardial infarction would choose TPA. Our hypothesis is that, for many informed individuals, regardless of cost, the increased risk of stroke may deter them from selecting TPA over SK. OBJECTIVE: To assess which thrombolytic drug informed patients would prefer and to explore the clinical and economic implications of such preferences. DESIGN: Prospective survey. SETTING: Tertiary care hospital. PATIENTS: 120 hospitalized patients with cardiac disease who would be "at risk" for a myocardial infarction. INTERVENTIONS: Face-to-face interviews utilizing a decision instrument. MEASUREMENTS: To minimize bias in soliciting patients' preferences and to standardize the presentation of information we developed a decision instrument which portrays a case scenario of a myocardial infarction, describes treatment outcomes (survival and stroke rate), and displays the likelihood of these outcomes with SK and TPA using three scenarios: a base stroke risk (all patients data), a lower stroke risk (<75 years old data), a higher stroke risk (>75 years old data). Outcome data were derived from the published literature (GUSTO study). RESULTS: When presented the overall results of the GUSTO study, 60/120 (50%) expressed a preference for SK. When presented the outcome data for the subgroups of patients <75 years old (lower stroke rate), 37/120 (31%) preferred SK. When presented the subgroup data for patients >75 years old (higher stroke risk), 67/120 (56%) preferred SK. CONCLUSIONS: Regardless of the scenario that individuals were presented with, a substantial proportion of individuals (31-56%) who could potentially require thrombolytic therapy chose SK over TPA. This study should be repeated in other settings to establish the generalizability of our results. Assuming that these results will be consistent, considering the patient's perspective has significant implications on clinical decision making as well as from an economic perspective. PMID- 11004416 TI - Which chronic conditions are associated with better or poorer quality of life? AB - The objective of the present study is to compare the QL of a wide range of chronic disease patients. Secondary analysis of eight existing data sets, including over 15,000 patients, was performed. The studies were conducted between 1993 and 1996 and included population-based samples, referred samples, consecutive samples, and/or consecutive samples. The SF-36 or SF-24 were employed as generic QL instruments. Patients who were older, female, had a low level of education, were not living with a partner, and had at least one comorbid condition, in general, reported the poorest level of QL. On the basis of rank ordering across the QL dimensions, three broad categories could be distinguished. Urogenital conditions, hearing impairments, psychiatric disorders, and dermatologic conditions were found to result in relatively favorable functioning. A group of disease clusters assuming an intermediate position encompassed cardiovascular conditions, cancer, endocrinologic conditions, visual impairments, and chronic respiratory diseases. Gastrointestinal conditions, cerebrovascular/neurologic conditions, renal diseases, and musculoskeletal conditions led to the most adverse sequelae. This categorization reflects the combined result of the diseases and comorbid conditions. If these results are replicated and validated in future studies, they can be considered in addition to information on the prevalence of the diseases, potential benefits of care, and current disease-specific expenditures. This combined information will help to better plan and allocate resources for research, training, and health care. PMID- 11004417 TI - Measuring quality of sedation in adult mechanically ventilated critically ill patients. the Vancouver Interaction and Calmness Scale. Sedation Focus Group. AB - There are no reliable, valid, and responsive scales to measure the quality of sedation in adult critically ill patients. Our objective was to develop a summated rating scale with these properties and to define the minimal clinically important difference (MCID). We developed and tested the scale in an 18-bed medical-surgical intensive care unit (ICU) (12-bed acute and 6-bed subacute unit). Following identification of relevant domains and item derivation, 116 observations were made on 38 patients; psychometric properties and interrater reliability were assessed to allow item reduction. The final scale consisted of two five-item subscales quantifying calmness and interaction along a continuum from 5 to 30 points. Interrater reliability was 0.89 and 0.90; internal consistency was 0. 95 for both subscales. To test construct validity, MCID, and responsiveness 302 observations were made on 54 patients. Construct validity: calmness score vs. need for further intervention to make the patient calm (R = 0.82, P < 0.001); interaction score discriminated between acute vs. subacute units, mean scores 15.28 +/- 8.26 vs. 23.54 +/- 7.42, mean difference 8.27 (95% CI - 10.32 to -6.22); MCID - 2.2 and 2.5 for the calmness and interaction subscales; Guyatt's responsiveness statistics - 1.4 and 2.3. The Vancouver Interaction and Calmness Scale (VICS) is reliable, valid, and responsive. PMID- 11004418 TI - Construction of the Patient-Oriented Prostate Utility Scale (PORPUS): a multiattribute health state classification system for prostate cancer. AB - Health status indexes, such as the EuroQol, consist of a health state classification system and a set of utility weights. Indexes measure quality of life using a 0-1 utility score. Utilities for outcomes in prostate cancer (PC) are of unique importance, but generic indexes do not represent PC outcomes (e.g., sexual, urinary, bowel dysfunction) well, and may not capture their full impact. As a step toward improved utility measurement, we constructed a classification system for PC. We generated items for each of six health domains and rated their importance using interviews with 10 clinical experts and 80 patients. Key concepts were selected for each domain using item importance weightings, and a set of predetermined criteria. Text was developed to express levels of severity within each domain. Experts and two additional groups of patients (n = 40, n = 96) evaluated textual clarity and endorsed the content validity of the instrument. The final system consists of 10 domains with 4-6 levels each. The content validity of the system was endorsed by patients and experts. In conjunction with a set of utility weights, it may be used to develop a health status index, to improve utility measurement in patients, and to serve as a short psychometric (nonutility) instrument. PMID- 11004419 TI - Choice of effect measure for epidemiological data. AB - The debate concerning the choice of effect measure for epidemiologic data has been renewed in the literature, and it suggests some continuing disagreement between the pertinent clinical and statistical criteria. In this article, some defining characteristics of the main choices of effect measure [risk difference (RD), relative risk (RR), and odds ratio (OR)] for binary data are presented and compared, with consideration of both the clinical and statistical perspectives. Relationships of these measures to the relative risk reduction (RRR) and number needed to treat (NNT) are also discussed. A numerical comparison of models of constant RD, RR, and OR is made to assess when and by how much they might differ in practice. Typically the models show only small numerical differences, unless extreme extrapolation is involved. The RD and RR models can predict impossible event rates, either less than zero or greater than 100%. Each measure has potential theoretical justification. RD and RR may enjoy some advantages for communication of risk, but OR may be preferred for data analysis. A clear distinction should be maintained between the objectives of data analysis and subsequent risk communication, and different effect measures may be needed for each. PMID- 11004420 TI - Comparison of ultrasound, radiography, and clinical examination in the diagnosis of acute maxillary sinusitis: a systematic review. AB - The objective of this study was to assess the discriminative properties of the methods for diagnosing acute maxillary sinusitis (AMS) in unselected patients. The study design was a systematic review of evaluation studies identified by using Medline, by searching reference lists, by hand searches, and by contacting investigators. Evaluation studies were conducted anywhere in the world. Subjects were adults with suspected AMS. Main outcome measures were: sensitivity, specificity, positive and negative likelihood ratios of the primary studies, weighted means of these parameters in each comparison (clinical examination, radiography, and ultrasound compared to a reference standard in diagnosing AMS), and summary ROC curves and their Q* points where sensitivity equals specificity. For the years from 1962 to present, 49 study reports were found; 11 articles on studies that included a total of 1144 patients were eligible. Compared to sinus puncture, radiography was the most accurate method for diagnosing AMS: the Q* point on the summary ROC curve was 0.82 (95% confidence interval, CI, 0.78-0.85). Ultrasound was slightly less accurate than radiography compared to sinus puncture (Q* 0.80, 95% CI 0.76-0.83). Only two articles reported clinical examination compared to sinus puncture and the Q* for them was 0.75 (95% 0.58-0.86). Clinical examination is a rather unreliable method for diagnosing AMS, even in the hands of experienced specialists. Using radiography or ultrasound improves the accuracy of diagnosis. The diagnosis of AMS is rarely studied in primary care settings. Future comparative trials should preferably combine diagnosis and treatment, evaluating the two aspects of clinical management as unit. PMID- 11004421 TI - Are clinical practice guidelines valid for primary care? AB - Development of evidence-based clinical practice guidelines has emphasized treatment efficacy under methodologically rigorous conditions; internal validity (i.e., efficacy) has been assessed well. Despite their experimental foundations, guidelines originally were considered physician education tools relevant to singular, idealized encounters. Now, in rushing to generalize, quantify, and regulate quality, guidelines are being applied to whole populations as quality review criteria. Clinical guidelines do not make operational how, or even make clear whether they should be so used. We studied the external validity, the generalizability and measurability for primary care, of a guideline for treatment of systolic heart failure. Patient and clinical factors reducing generalizability were examined. Imprecise definition and reliance on single measures of recommendations were studied as limits to measurability. Patient, physician, organizational, and system factors influenced guideline validity. Attention to myriad invalidity sources undermining implementation and evaluation of efforts to improve quality and outcomes of primary care is critical. PMID- 11004422 TI - Education, aging, and health: to what extent can the rise in educational level relieve the future health (care) burden associated with population aging in the Netherlands? AB - This article describes to what extent the expected rise in the educational level of the Dutch population can counterbalance the increases in the prevalence of ill health and health care utilization based on the aging of the population for the period 1996-2020. Logistic regression models are used to estimate current differences in health (care utilization) by age, sex, and educational level, using data from the Netherlands Health Interview Survey. The current differences in health (care utilization) are applied to national projections of the composition of the population by age, sex, and educational level. Also, scenarios have been made in which the health differences by educational level are assumed to converge and diverge. The rise in the educational level counteracts the expected increases in ill-health based on population aging to a substantial degree (10-100%). We therefore recommend that in projections of ill-health also changes in educational level are taken into account. PMID- 11004423 TI - What contributions do languages other than English make on the results of meta analyses? AB - Including only a portion of all available evidence may introduce systematic errors into the meta-analytic process and threaten its validity. We set out to examine whether language restricted meta-analyses, compared to language inclusive meta-analyses, provide different estimates of the effectiveness of interventions evaluated in randomized trials. We identified and retrieved all 79 meta-analyses from several disease areas in which explicit eligibility criteria regarding trial selection were reported. General characteristics and quality of reporting of the meta-analyses were assessed using a validated instrument. We explored the effects of language of publication of the randomized trials on the quantitative results using logistic regression analyses. Language restricted meta-analyses, compared to language inclusive meta-analyses, did not differ with respect to the estimate of benefit of the effectiveness of an intervention (ROR = 0.98; 95% CI: 0.81 1.17). These results were also robust after a series of sensitivity analyses. This study provides no evidence that language restricted meta-analyses lead to biased estimates of intervention effectiveness. We encourage others to replicate this study using different sampling frames, clinical topics and interventions. PMID- 11004424 TI - New insights into metabolic and vascular management of type 2 diabetes. Proceedings of a meeting. Brussels, Belgium, September 1999. PMID- 11004425 TI - Therapeutic objectives and their practical achievement in type 2 diabetes. AB - This paper offers a broad but selective account of the context of type 2 diabetes at the start of the new millennium. It outlines the major long-term threats to health and life of people with type 2 diabetes and indicates that, while their relative impacts may differ, the burden of the specific "microvascular" and the atherosclerotic "macrovascular" complications of diabetes weigh as heavily on people with type 2 as on those with type 1 diabetes. Correction of hyperglycemia still has a leading role among therapeutic objectives in type 2 diabetes, but the concept of the "defence in depth" and the crucial importance of control of arterial pressure and correction of dyslipidemia - the "bad companions" in diabetes - is stressed. Other defences against tissue and organ failure in diabetes are described, highlighting the importance of regular, systematic screening for risk factors and early manifestations of tissue damage. Broader organizational, social and economic factors in diabetes care are touched upon and the need for strong alliances of all concerned parties - care providers, managers, health politicians, and above all informed and motivated people with diabetes themselves - is underlined with reference to the World Health Organization/International Diabetes Federation St. Vincent Declaration Initiative in Europe and its recent 10-year anniversary "Istanbul Commitment" to full implementation of the Declaration. PMID- 11004426 TI - Diamicron MR once daily is effective and well tolerated in type 2 diabetes: a double-blind, randomized, multinational study. AB - In a double-blind, multicenter, multinational study, the long-term efficacy and safety of Diamicron(R)MR, a new gliclazide formulation taken once daily at lower dose (30-120 mg/day) was compared with Diamicron(R) (80-320 mg/day) taken twice daily in type 2 diabetic outpatients. After a 2-week run-in period, 800 patients with poor blood glucose control were randomized to Diamicron(R)MR (n=401) or Diamicron(R) (n=399). After a 4-month titration period, the efficacy and safety of Diamicron(R)MR was compared with Diamicron(R) over a 6-month fixed-dose treatment period. The ability to switch from Diamicron(R) to Diamicron(R)MR was then assessed during an additional 2-month follow-up period. Equivalence between treatment with Diamicron(R)MR and Diamicron(R) was compared by a non-inferiority test; the limit of equivalence was set at 0.5% for HbA(1c) and 1 mmol/l for fasting plasma glucose (FPG). The treatment groups were comparable at baseline. After 10 months of treatment, Diamicron(R)MR was as efficient as Diamicron(R) in controlling blood glucose, with a mean end point difference in HbA(1c) of -0.08 (0. 08)%, significantly lower than the equivalence limit (p<0.001). Similar results were obtained for FPG.The safety of Diamicron(R)MR and Diamicron(R) was equally high. The incidence of hypoglycemia was particularly low (0.2 hypoglycemia/100 patient months) in the elderly population, which represented almost 40% of the included patients. This study demonstrates that 30 to 120 mg of Diamicron(R)MR taken once daily is at least as efficient as 80 to 320 mg of Diamicron(R) taken in divided doses with respect to HbA(1c) and FPG levels, with a similar safety profile. PMID- 11004427 TI - Tissue-specific effects of sulfonylureas: lessons from studies of cloned K(ATP) channels. AB - Sulfonylureas stimulate insulin secretion in type-2 diabetic patients by blocking ATP-sensitive (K(ATP)) potassium channels in the pancreatic beta-cell membrane. This effect is mediated by the binding of the drug to the sulfonylurea receptor (SUR) subunit of the channel. K(ATP) channels are also present in other tissues, but often contain different types of SUR subunits (e.g., SUR1 in beta-cells, SUR2A in heart, SUR2B in smooth muscle). The sensitivity of these different types of K(ATP) channels to sulfonylureas is variable: gliclazide and tolbutamide block the beta-cell, but not the cardiac or smooth muscle, types of K(ATP) channel. In contrast, glibenclamide blocks all three types of channel with similar affinity. The reversibility of the drugs also varies, with tolbutamide and gliclazide being reversible on all three types of K(ATP) channel, while glibenclamide is reversible on cardiac, but not beta-cell, K(ATP) channels. This review summarizes current knowledge of how sulfonylureas act on the different types of K(ATP) channel found in beta-cells and in extrapancreatic tissues, and discusses the implications of these findings for their use as therapeutic agents. PMID- 11004428 TI - Possible pathophysiological mechanisms for diabetic angiopathy in type 2 diabetes. AB - The expression of large and small vessel disease in type 2 diabetes differs from that observed in type 1, with a higher prevalence of atherosclerosis and hypertension, maculopathy rather than proliferative retinopathy, and nephropathy of a more complex nature. Such differences are mirrored by differences in vascular pathophysiology with an early impairment of microvascular vasodilatory reserve being a prominent feature. The defect appears to be endothelium dependent and in conjunction with evidence of endothelium activation suggests that the endothelium plays a crucial role in the pathogenesis of vascular disease in type 2 diabetes and may even be an intrinsic feature or common antecedent of the insulin resistance syndrome. Several cellular mechanisms may be proposed linking insulin resistance and endothelial dysfunction including (i) abnormalities of common signal transduction mechanisms, (ii) alterations in cell membrane fluidity altering the expression and/or presentation of a wide range of receptors, or (iii) changes in oxidative stress. It is intuitively unlikely that the alteration of a single signal transduction mechanism could be a common cause, particularly as aspects of endothelial dysfunction implicate different mechanisms. Accordingly, changes in oxidative stress, either stemming from glucose-mediated increased free-radical generation and/or reduction of antioxidant capacity, are strong contender mechanisms. Not only may increased oxidative stress result in the quenching of nitric oxide, neutralizing its many protective functions, but it may also damage DNA, protein structure, and membrane properties. Elucidating the links between oxidative stress, endothelial function, and insulin resistance has important implications for the prevention of diabetic angiopathy and perhaps for the prevention of diabetes itself. PMID- 11004429 TI - In vitro and in vivo antioxidant properties of gliclazide. AB - Diabetes is a state of increased oxidant stress and there is evidence that oxidation may play a role in the genesis of complications. Gliclazide, a sulfonylurea hypoglycemic drug, has been shown to possess free radical scavenging properties. This study examined the effects of in vitro supplementation with gliclazide and other sulfonylureas as on low-density lipoprotein (LDL) oxidation and the total plasma antioxidant capacity (TPAC). In a separate study, the effects of 10 months of oral gliclazide therapy on oxidative parameters were assessed in 44 type 2 diabetic patients. Gliclazide, but not glibenclamide, glimepiride, glipizide or tolbutamide, inhibited LDL oxidation and enhanced TPAC. With the addition of 1 microM gliclazide, oxidation lag time increased from 53.6+/-2.6 to 113.6+/-5.1 min (p<0.001), and TPAC increased from 1. 09+/-0.11 to 1.23+/-0.11 mM (p<0.01). Administration of either modified release or standard gliclazide to type 2 diabetic patients resulted in a fall in 8-isoprostanes, a marker of lipid oxidation, and an increase in the antioxidant parameters TPAC, SOD and thiols. These studies show that gliclazide possesses antioxidant properties that produce measurable clinical effects at therapeutic doses. PMID- 11004430 TI - Correction of glycosylated oxyhemoglobin-induced impairment of endothelium dependent vasodilatation by gliclazide. AB - We have investigated whether gliclazide, a second-generation sulfonylurea hypoglycemic agent, interferes with the impairment of endothelium-dependent nitric-oxide-mediated relaxation produced by 14%-glycosylated human oxyhemoglobin (GHHb). For comparative purposes, other agents, like glibenclamide, aminoguanidine, ascorbic acid or superoxide dismutase (SOD), were also tested. GHHb (10 nM) caused a reduction in endothelium-dependent relaxation induced by acetylcholine (1 nM to 10 microM) in both isolated aortic segments and mesenteric microvessels from normoglycemic nondiabetic rats. Preincubation of the vessels with gliclazide (100 nM to 10 microM) prevented the impairment of endothelial relaxation, the threshold concentration of gliclazide being 300 nM. In addition, 10 microM gliclazide also prevented the reduction by 10 nM GHHb of the relaxation induced by exogenous nitric oxide (NO, 10 nM to 100 microM). Determination of superoxide anion release measured by the reduction in ferricytochrome c indicated that GHHb produced significant amounts of these free radicals that were concentration-dependently inhibited by gliclazide. The impairment of endothelium mediated responses was also prevented by 100 U/ml SOD or 10 microM ascorbic acid, but not by 10 microM glibenclamide or 100 microM aminoguanidine. We conclude that gliclazide can reduce the impairment of nitric-oxide-mediated endothelium dependent relaxation produced by GHHb. This reduction is likely related to the antioxidant properties of the drug, a mechanism suggested by these studies which demonstrate the inactivation of superoxide anions produced by the glycosylated protein by gliclazide. PMID- 11004431 TI - Effect of gliclazide on monocyte-endothelium interactions in diabetes. AB - Enhanced monocyte-endothelial cell interactions have been documented in diabetes. Because adherence of monocytes to the endothelium is one of the earliest events in the development of atherosclerosis, its alteration may represent one of the mechanisms leading to accelerated atherosclerosis in diabetic patients. Previous studies have suggested that lipoprotein oxidation and protein glycation may contribute to the increased monocyte binding to the diabetic vasculature. Based on the recent finding that gliclazide has free-radical scavenging activity, we examined the ex vivo and in vitro effects of this drug on human monocyte binding to endothelial cells. Our results demonstrate that short-term administration of gliclazide to patients with type 2 diabetes lowers the enhanced adhesion of diabetic monocytes observed before gliclazide treatment (163+/-24% over control values, p<0.005) to levels similar to those observed in controls. They also show that gliclazide (10 microg/ml) reduces in vitro by approximately 35% both oxidized low-density lipoprotein (LDL)- and glycated albumin-induced monocyte adhesion to endothelial cells. Based on these results, we next investigated the molecular mechanisms responsible for the inhibitory effect of gliclazide on glycated albumin-induced monocyte adhesion to endothelium. In glycated albumin treated endothelial cells, we observed induction of cell-associated expression of E-selectin (ELAM-1; 170+/-10% over control values, p<0.005), intercellular cell adhesion molecule-1 (ICAM-1; 131+/-8% over control values, p<0.005) and vascular cell adhesion molecule-1 (VCAM-1; 134+/-8% over control values, p<0.005), augmentation in the levels of the transcripts of these molecules, and an increase in the DNA binding of NF-kappaB in the promoters of these antigens. Gliclazide markedly inhibited the induction of all these parameters. Because the oxidative stress-sensitive transcription factor NF-kappaB is implicated in endothelial cell activation, the observed inhibitory effect of gliclazide on NF-kappaB activation and glycated albumin-induced expression of DNA binding activity for the NF-kappaB site in the ELAM-1, ICAM-1 and VCAM-1 promoters seems to be due to its antioxidant properties. These results suggest that gliclazide, by its ability to reduce endothelial activation, may exert potential beneficial effects in the prevention of atherosclerosis associated with type 2 diabetes. PMID- 11004432 TI - Prevention of endothelial dysfunction in streptozotocin-induced diabetic rats by gliclazide treatment. AB - The aim of the present work was to analyze whether the oral hypoglycemic drug gliclazide affects diabetic endothelial dysfunction in streptozotocin-induced diabetic rats. Gliclazide was compared with glibenclamide, ascorbic acid, and aminoguanidine. An insulin-dependent model of diabetes was selected to exclude insulin-releasing effects of the drugs. Both in isolated aortic segments and mesenteric microvessels, endothelium-dependent relaxation evoked by acetylcholine (ACh, 1 nM to 10 microM) was significantly reduced in vessels from diabetic animals. This impairment was reversed when the segments were previously incubated with 100 U/ml superoxide dismutase. When streptozotocin-induced diabetic rats were orally treated from the time of diabetes induction with gliclazide (10 mg/kg) or ascorbic acid (250 mg/kg), ACh-induced endothelium-dependent relaxation was well preserved both in aortic segments and mesenteric microvessels. In addition, the impaired vasodilatation to exogenous nitric oxide (NO) in aortic segments was also improved in gliclazide-treated diabetic rats. On the other hand, oral treatment with glibenclamide (1 and 10 mg/kg) or aminoguanidine (250 mg/kg) did not produce significant improvements in diabetic endothelial dysfunction. We conclude that gliclazide reverses the endothelial dysfunction associated with diabetes. This effect appears to be due not to the metabolic actions of the drug but rather to its antioxidant properties, as it can be mimicked by other antioxidants. We propose that the mechanism involved is the inactivation of reactive oxygen species, which are increased in diabetes probably as a result of increased early protein glycosylation products, such as glycosylated hemoglobin (HbA(1c)). These effects of gliclazide are not shared by other oral hypoglycemic agent such as glibenclamide, or by blockade of advanced glycosylation end product (AGE) generation with aminoguanidine. PMID- 11004433 TI - Excitation energy transfer from phycobilisomes to photosystems: a phenomenon associated with the temporal separation of photosynthesis and nitrogen fixation in a cyanobacterium, Plectonema boryanum(1). AB - Plectonema boryanum shows temporal separation of photosynthesis and nitrogen fixation under diazotrophic conditions. Low temperature fluorescence studies have shown that in vivo the nitrogen fixing and photosynthesizing cells are adapted to 'state 2' and 'state 1', respectively. During nitrogen fixation phycobilisomes seem to transfer excitation energy to photosystem I whereas during oxygenic photosynthesis the energy is transferred to photosystem II. The state 2 adapted N phase cells failed to undergo transition to state 1 while P-phase cells exhibited state 1 to state 2 transition. The nitrogen fixing cells showed a decreased level of psbC transcript, lack of CP47 in thylakoid membrane, and presence of the F685 peak but absence of the F695 peak in 77 K fluorescence spectra. These results suggest that the metabolic and molecular changes associated with nitrogen fixation may favor direct energy transfer from the phycobilisomes to photosystem I. This should help the organism to achieve low photosystem II and high photosystem I activity to set temporal separation of nitrogen fixation and photosynthesis for photoautotrophic growth under diazotrophic conditions. PMID- 11004434 TI - Conformational relaxation following reduction of the photoactive bacteriopheophytin in reaction centers from Blastochloris viridis. Influence of mutations at position M208. AB - The photochemically trapped bacteriopheophytin (BPh) b radical anion in the active branch (Phi(A)(&z.rad;-)) of reaction centers (RCs) from Blastochloris (formerly called Rhodopseudomonas) viridis is characterized by 1H-ENDOR as well as optical absorption spectroscopy. The two site-directed mutants YF(M208) and YL(M208), in which tyrosine at position M208 is replaced by phenylalanine and leucine, respectively, are investigated and compared with the wild type. The residue at M208 is in close proximity to the primary electron donor, P, the monomeric bacteriochlorophyll (BChl), B(A), and the BPh, Phi(A), that are involved in the transmembrane electron transfer to the quinone, Q(A), in the RC. The analysis of the ENDOR spectra of Phi(A)(&z.rad;-) at 160 K indicates that two distinct states of Phi(A)(&z.rad;-) are present in the wild type and the mutant YF(M208). Based on a comparison with Phi(A)(&z.rad;-) in RCs of Rhodobacter sphaeroides the two states are interpreted as torsional isomers of the 3-acetyl group of Phi(A). Only one Phi(A)(&z.rad;-) state occurs in the mutant YL(M208). This effect of the leucine residue at position M208 is explained by steric hindrance that locks the acetyl group in one specific position. On the basis of these results, an interpretation of the optical absorption difference spectrum of the state Phi(A)(&z.rad;-)Q(A)(&z.rad;-) is attempted. This state can be accumulated at 100 K and undergoes an irreversible change between 100 and 200 K [Tiede et al., Biochim. Biophys. Acta 892 (1987) 294-302]. The corresponding absorbance changes in the BChl Q(x) and Q(y) regions observed in the wild type also occur in the YF(M208) mutant but not in YL(M208). The observed changes in the wild type and YF(M208) are assigned to RCs in which the 3-acetyl group of Phi(A) changes its orientation. It is concluded that this distinct structural relaxation of Phi(A) can significantly affect the optical properties of B(A) and contribute to the light-induced absorption difference spectra. PMID- 11004435 TI - Covalent modification of the catalytic sites of the H(+)-ATPase from chloroplasts with 2-nitreno-ADP. Modification of the catalytic site 1 (tight) and catalytic sites 1 and 2 together impairs both uni-site and multi-site catalysis of ATP synthesis and ATP hydrolysis. AB - After isolation and purification, the H(+)-ATPase from chloroplasts, CF(0)F(1), contains one endogenous ADP at a catalytic site, and two endogenous ATP at non catalytic sites. Incubation with 2-azido-[alpha-(32)P]ADP leads to tight binding of azido-nucleotides. Free nucleotides were removed by three consecutive passages through centrifugation columns, and upon UV-irradiation most of the label was covalently bound. The labelled enzyme was digested by trypsin, the peptides were separated by ion exchange chromatography into nitreno-AMP, nitreno-ADP and nitreno-ATP labelled peptides, and these were then separated by reversed phase chromatography. Amino acid sequence analysis was used to identify the type of the nucleotide binding site. After incubation with 2-azido-[alpha-(32)P]ADP, the covalently bound label was found exclusively at beta-Tyr-362. Incubation conditions with 2-azido-[alpha-(32)P]ADP were varied, and conditions were found which allow selective binding of the label to different catalytic sites, designated as 1, 2 and 3 in order of decreasing affinity for ADP, and either catalytic site 1 or catalytic sites 1 and 2 together were labelled. For measurements of the degree of inhibition by covalent modification, CF(0)F(1) was reconstituted into phosphatidylcholine liposomes, and the membranes were energised by an acid-base transition in the presence of a K(+)/valinomycin diffusion potential. The rate of ATP synthesis was 50-80 s(-1), and the rate of ATP hydrolysis was 15 s(-1) measured under multi-site conditions. Covalent modification of either catalytic site 1 or catalytic sites 1 and 2 together inhibited ATP synthesis and ATP hydrolysis equally, the degree of inhibition being proportional to the degree of modification. Extrapolation to complete inhibition indicates that derivatisation of catalytic site 1 leads to complete inhibition when 1 mol 2-nitreno-ADP is bound per mol CF(0)F(1). Derivatisation of catalytic sites 1 and 2 together extrapolates to complete inhibition when 2 mol 2 nitreno-ADP are bound per CF(0)F(1). The rate of ATP synthesis and the rate of ATP hydrolysis were measured as a function of the substrate concentration from multi-site to uni-site conditions with derivatised CF(0)F(1) and with non derivatised CF(0)F(1). ATP synthesis and ATP hydrolysis under uni-site and under multi-site condition were inhibited by covalent modification of either catalytic site 1 or catalytic sites 1 and 2 together. The results indicate that derivatisation of site 1 inhibits activation of the enzyme and that cooperative interactions occur at least between the catalytic sites 2 and 3. PMID- 11004436 TI - Revealing the structure of the photosystem II chlorophyll binding proteins, CP43 and CP47. AB - A review of the structural properties of the photosystem II chlorophyll binding proteins, CP47 and CP43, is given and a model of the transmembrane helical domains of CP47 has been constructed. The model is based on (i) the amino acid sequence of the spinach protein, (ii) an 8 A three-dimensional electron density map derived from electron crystallography and (iii) the structural homology which the membrane spanning region of CP47 shares with the six N-terminal transmembrane helices of the PsaA/PsaB proteins of photosystem I. Particular emphasis has been placed on the position of chlorophyll molecules assigned in the 8 A three dimensional map of CP47 (K.-H. Rhee, E.P. Morris, J. Barber, W. Kuhlbrandt, Nature 396 (1998) 283-286) relative to histidine residues located in the transmembrane regions of this protein which are likely to form axial ligands for chlorophyll binding. Of the 14 densities assigned to chlorophyll, the model predicted that five have their magnesium ions within 4 A of the imidazole nitrogens of histidine residues. For the remaining seven histidine residues the densities attributed to chlorophylls were within 4-8 A of the imidazole nitrogens and thus too far apart for direct ligation with the magnesium ion within the tetrapyrrole head group. Improved structural resolution and reconsiderations of the orientation of the porphyrin rings will allow further refinement of the model. PMID- 11004437 TI - Solution structure of the NADP(H)-binding component (dIII) of proton translocating transhydrogenase from Rhodospirillum rubrum. AB - Transhydrogenase is a proton pump found in the membranes of bacteria and animal mitochondria. The solution structure of the expressed, 21.5 kDa, NADP(H)-binding component (dIII) of transhydrogenase from Rhodospirillum rubrum has been solved by NMR methods. This is the first description of the structure of dIII from a bacterial source. The protein adopts a Rossmann fold: an open, twisted, parallel beta-sheet, flanked by helices. However, the binding of NADP(+) to dIII is profoundly different to that seen in other Rossmann structures, in that its orientation is reversed: the adenosine moiety interacts with the first betaalphabetaalphabeta motif, and the nicotinamide with the second. Features in the structure that might be responsible for changes in nucleotide-binding affinity during catalysis, and for interaction with other components of the enzyme, are identified. The results are compared with the recently determined, high-resolution crystal structures of human and bovine dIII which also show the reversed nucleotide orientation. PMID- 11004438 TI - Application of the obligate aerobic yeast Yarrowia lipolytica as a eucaryotic model to analyse Leigh syndrome mutations in the complex I core subunits PSST and TYKY. AB - We have used the obligate aerobic yeast Yarrowia lipolytica to reconstruct and analyse three missense mutations in the nuclear coded subunits homologous to bovine TYKY and PSST of mitochondrial complex I (proton translocating NADH:ubiquinone oxidoreductase) that have been shown to cause Leigh syndrome (MIM 25600), a severe progressive neurodegenerative disorder. While homozygosity for a V122M substitution in NDUFS7 (PSST) has been found in two siblings with neuropathologically proven Leigh syndrome (R. Triepels et al., Ann. Neurol. 45 (1999) 787), heterozygosity for a P79L and a R102H substitution in NDUFS8 (TYKY) has been found in another patient (J. Loeffen et al., Am. J. Hum. Genet. 63 (1998) 1598). Mitochondrial membranes from Y. lipolytica strains carrying any of the three point mutations exhibited similar complex I defects, with V(max) being reduced by about 50%. This suggests that complex I mutations that clinically present as Leigh syndrome may share common characteristics. In addition changes in the K(m) for n-decyl-ubiquinone and I(50) for hydrophobic complex I inhibitors were observed, which provides further evidence that not only the hydrophobic, mitochondrially coded subunits, but also some of the nuclear coded subunits of complex I are involved in its reaction with ubiquinone. PMID- 11004439 TI - Nitric oxide reductases in bacteria. AB - Nitric oxide reductases (NORs) that are found in bacteria belong to the large enzyme family which includes cytochrome oxidases. Two types of bacterial NORs have been characterised. One is a cytochrome bc-type complex (cNOR) that receives electrons from soluble redox protein donors, whereas the other type (qNOR) lacks the cytochrome c component and uses quinol as the electron donor. The latter enzyme is present in several pathogens that are not denitrifiers. We summarise the current knowledge on bacterial NORs, and discuss the evolutionary relationship between them and cytochrome oxidases in this review. PMID- 11004440 TI - Diversity and origin of alternative NADH:ubiquinone oxidoreductases. AB - Mitochondria from various organisms, especially plants, fungi and many bacteria contain so-called alternative NADH:ubiquinone oxidoreductases that catalyse the same redox reaction as respiratory chain complex I, but do not contribute to the generation of transmembrane proton gradients. In eucaryotes, these enzymes are associated with the mitochondrial inner membrane, with their NADH reaction site facing either the mitochondrial matrix (internal alternative NADH:ubiquinone oxidoreductases) or the cytoplasm (external alternative NADH:ubiquinone oxidoreductases). Some of these enzymes also accept NADPH as substrate, some require calcium for activity. In the past few years, the characterisation of several alternative NADH:ubiquinone oxidoreductases on the DNA and on the protein level, of substrate specificities, mitochondrial import and targeting to the mitochondrial inner membrane has greatly improved our understanding of these enzymes. The present review will, with an emphasis on yeast model systems, illuminate various aspects of the biochemistry of alternative NADH:ubiquinone oxidoreductases, address recent developments and discuss some of the questions still open in the field. PMID- 11004441 TI - The transmembrane domain and the proton channel in proton-pumping transhydrogenases. AB - Proton-pumping nicotinamide nucleotide transhydrogenases are composed of three main domains, the NAD(H)-binding and NADP(H)-binding hydrophilic domains I (dI) and III (dIII), respectively, and the hydrophobic domain II (dII) containing the assumed proton channel. dII in the Escherichia coli enzyme has recently been characterised with regard to topology and a packing model of the helix bundle in dII is proposed. Extensive mutagenesis of conserved charged residues of this domain showed that important residues are betaHis91 and betaAsn222. The pH dependence of betaH91D, as well as betaH91C (unpublished), when compared to that of wild type shows that reduction of 3-acetylpyridine-NAD(+) by NADPH, i.e., the reverse reaction, is optimal at a pH essentially coinciding with the pK(a) of the residue in the beta91 position. It is therefore concluded that the wild-type transhydrogenase is regulated by the degree of protonation of betaHis91. The mechanisms of the interactions between dI+dIII and dII are suggested to involve pronounced conformational changes in a 'hinge' region around betaR265. PMID- 11004442 TI - Mitochondrial electron transfer in the wheat pathogenic fungus Septoria tritici: on the role of alternative respiratory enzymes in fungicide resistance. AB - Certain phytopathogenic fungi are able to express alternative NADH- and quinol oxidising enzymes that are insensitive to inhibitors of the mitochondrial respiratory Complexes I and III. To assess the extent to which such enzymes confer tolerance to respiration-targeted fungicides, an understanding of mitochondrial electron transfer in these species is required. An isolation procedure has been developed which results in intact, active and coupled mitochondria from the wheat pathogen Septoria tritici, as evidenced by morphological and kinetic data. Exogenous NADH, succinate and malate/glutamate are readily oxidised, the latter activity being only partly (approx. 70%) sensitive to rotenone. Of particular importance was the finding that azoxystrobin (a strobilurin fungicide) potently inhibits fungal respiration at the level of Complex III. In some S. tritici strains investigated, a small but significant part of the respiratory activity (approx. 10%) is insensitive to antimycin A and azoxystrobin. Such resistant activity is sensitive to octyl gallate, a specific inhibitor of the plant alternative oxidase. This enzyme, however, could not be detected immunologically. On the basis of the above findings, a conceptual mitochondrial electron transfer chain is presented. Data are discussed in terms of developmental and environmental regulation of the composition of this chain. PMID- 11004443 TI - Characterization of the complex I-associated ubisemiquinone species: toward the understanding of their functional roles in the electron/proton transfer reaction. AB - NADH-ubiquinone oxidoreductase (called complex I for mitochondrial enzyme and NDH 1 for bacterial counterparts) is an energy transducer, which utilizes the redox energy derived from the oxidation of NADH with ubiquinone to generate an electrochemical proton gradient (Deltamu(H(+))) across the membrane. The complex I/NDH-1 contain one non-covalently bound flavin mononucleotide and as many as eight iron-sulfur clusters as electron transfer components in common. In addition, electron paramagnetic resonance (EPR) spectroscopic studies have revealed that three ubisemiquinone (SQ) species with distinct spectroscopic and thermodynamic properties are detectable in complex I and function as electron/proton translocators. Thus, the understanding of molecular properties of the individual quinone species is prerequisite to elucidate the energy-coupling mechanism of complex I. We have investigated these SQ species using EPR spectroscopy and found that the three SQ species have strikingly different properties. We will report characteristics of these SQ species and discuss possible functional roles of individual quinone species in the electron/proton transfer reaction of complex I/NDH-1. PMID- 11004444 TI - Characterization of two novel redox groups in the respiratory NADH:ubiquinone oxidoreductase (complex I). AB - The proton-pumping NADH:ubiquinone oxidoreductase is the first of the respiratory chain complexes in many bacteria and mitochondria of most eukaryotes. The bacterial complex consists of 14 different subunits. Seven peripheral subunits bear all known redox groups of complex I, namely one FMN and five EPR-detectable iron-sulfur (FeS) clusters. The remaining seven subunits are hydrophobic proteins predicted to fold into 54 alpha-helices across the membrane. Little is known about their function, but they are most likely involved in proton translocation. The mitochondrial complex contains in addition to the homologues of these 14 subunits at least 29 additional proteins that do not directly participate in electron transfer and proton translocation. A novel redox group has been detected in the Neurospora crassa complex, in an amphipathic fragment of the Escherichia coli complex I and in a related hydrogenase and ferredoxin by means of UV/Vis spectroscopy. This group is made up by the two tetranuclear FeS clusters located on NuoI (the bovine TYKY) which have not been detected by EPR spectroscopy yet. Furthermore, we present evidence for the existence of a novel redox group located in the membrane arm of the complex. Partly reduced complex I equilibrated to a redox potential of -150 mV gives a UV/Vis redox difference spectrum that cannot be attributed to the known cofactors. Electrochemical titration of this absorption reveals a midpoint potential of -80 mV. This group is believed to transfer electrons from the high potential FeS cluster to ubiquinone. PMID- 11004445 TI - Catalysis in fumarate reductase. AB - In the absence of oxygen many bacteria are able to utilise fumarate as a terminal oxidant for respiration. In most known organisms the fumarate reductases are membrane-bound iron-sulfur flavoproteins but Shewanella species produce a soluble, periplasmic flavocytochrome c(3) that catalyses this reaction. The active sites of all fumarate reductases are clearly conserved at the structural level, indicating a common mechanism. The structures of fumarate reductases from two Shewanella species have been determined. Fumarate, succinate and a partially hydrated fumarate ligand are found in equivalent locations in different crystals, tightly bound in the active site and close to N5 of the FAD cofactor, allowing identification of amino acid residues that are involved in substrate binding and catalysis. Conversion of fumarate to succinate requires hydride transfer from FAD and protonation by an active site acid. The identity of the proton donor has been open to question but we have used structural considerations to suggest that this function is provided by an arginine side chain. We have confirmed this experimentally by analysing the effects of site-directed mutations on enzyme activity. Substitutions of Arg402 lead to a dramatic loss of activity whereas neither of the two active site histidine residues is required for catalysis. PMID- 11004446 TI - Haem-polypeptide interactions during cytochrome c maturation. AB - Cytochrome c maturation involves the translocation of a polypeptide, the apocytochrome, and its cofactor, haem, through a membrane, before the two molecules are ligated covalently. This review article focuses on the current knowledge on the journey of haem during this process, which is known best in the Gram-negative bacterium Escherichia coli. As haem always occurs bound to protein, its passage across the cytoplasmic membrane and incorporation into the apocytochrome appears to be mediated by a set of proteinaceous maturation factors, the Ccm (cytochrome c maturation) proteins. At least three of them, CcmC, CcmE and CcmF, are thought to interact directly with haem. CcmE binds haem covalently, thus representing an intermediate of the haem trafficking pathway. CcmC is required for binding of haem to CcmE, and CcmF for releasing it from CcmE and transferring it onto the apocytochrome. The mechanism by which haem crosses the cytoplasmic membrane is currently unknown. PMID- 11004447 TI - A novel protein transport system involved in the biogenesis of bacterial electron transfer chains. AB - The Tat system is a recently discovered bacterial protein transport pathway that functions primarily in the biosynthesis of proteins containing redox active cofactors. Analogous transport systems are found in plant organelles. Remarkably and uniquely the Tat system functions to transported a diverse range of folded proteins across a biological membrane, a feat that must be achieved without rendering the membrane freely permeable to protons and other ions. Here we review the operation of the bacterial Tat system and propose a model for the structural organisation of the Tat preprotein translocase. PMID- 11004448 TI - What fuels polypeptide translocation? An energetical view on mitochondrial protein sorting. AB - Protein sorting into mitochondria is achieved by the concerted action of at least four translocation complexes. Vectorial transport of polypeptide chains by these complexes requires different driving forces. In particular, Deltapsi, matrix adenosine triphosphate and the free energy of the binding to other protein components are used in series to achieve sorting of proteins to the various mitochondrial subcompartments. The processes providing the translocation energy are presented in this review and their impact for protein sorting into and within mitochondria is discussed. PMID- 11004449 TI - Crystallographic studies of the conformational changes that drive directional transmembrane ion movement in bacteriorhodopsin. AB - Recent advances in the determination of the X-ray crystallographic structures of bacteriorhodopsin, and some of its photointermediates, reveal the nature of the linkage between the relaxation of electrostatic and steric conflicts at the retinal and events elsewhere in the protein. The transport cycle can be now understood in terms of specific and well-described displacements of hydrogen bonded water, and main-chain and side-chain atoms, that lower the pK(a)s of the proton release group in the extracellular region and Asp-96 in the cytoplasmic region. Thus, local electrostatic conflict of the photoisomerized retinal with Asp-85 and Asp-212 causes deprotonation of the Schiff base, and results in a cascade of events culminating in proton release to the extracellular surface. Local steric conflict of the 13-methyl group with Trp-182 causes, in turn, a cascade of movements in the cytoplasmic region, and results in reprotonation of the Schiff base. Although numerous questions concerning the mechanism of each of these proton (or perhaps hydroxyl ion) transfers remain, the structural results provide a detailed molecular explanation for how the directionality of the ion transfers is determined by the configurational relaxation of the retinal. PMID- 11004450 TI - Electron transfer during the oxidation of ammonia by the chemolithotrophic bacterium Nitrosomonas europaea. AB - The combined action of ammonia monooxygenase, AMO, (NH(3)+2e(-)+O(2)-->NH(2)OH) and hydroxylamine oxidoreductase, HAO, (NH(2)OH+H(2)O-->HNO(2)+4e(-)+4H(+)) accounts for ammonia oxidation in Nitrosomonas europaea. Pathways for electrons from HAO to O(2), nitrite, NO, H(2)O(2) or AMO are reviewed and some recent advances described. The membrane cytochrome c(M)552 is proposed to participate in the path between HAO and ubiquinone. A bc(1) complex is shown to mediate between ubiquinol and the terminal oxidase and is shown to be downstream of HAO. A novel, red, low-potential, periplasmic copper protein, nitrosocyanin, is introduced. Possible mechanisms for the inhibition of ammonia oxidation in cells by protonophores are summarized. Genes for nitrite- and NO-reductase but not N(2)O or nitrate reductase are present in the genome of Nitrosomonas. Nitrite reductase is not repressed by growth on O(2); the flux of nitrite reduction is controlled at the substrate level. PMID- 11004451 TI - Mammalian mitochondrial inner membrane cationic and neutral amino acid carriers. PMID- 11004452 TI - Identification and functions of new transporters in yeast mitochondria. AB - The genome of Saccharomyces cerevisiae encodes 35 putative members of the mitochondrial carrier family. Known members of this family transport substrates and products across the inner membranes of mitochondria. We are attempting to identify the functions of the yeast mitochondrial transporters via high-yield expression in Escherichia coli and/or S. cerevisiae, purification and reconstitution of their protein products into liposomes, where their transport properties are investigated. With this strategy, we have already identified the functions of seven S. cerevisiae gene products, whose structural and functional properties assigned them to the mitochondrial carrier family. The functional information obtained in the reconstituted system and the use of knock-out yeast strains can be usefully exploited for the investigation of the physiological role of individual transporters. Furthermore, the yeast carrier sequences can be used to identify the orthologous proteins in other organisms, including man. PMID- 11004453 TI - Biogenesis of iron-sulfur proteins in eukaryotes: a novel task of mitochondria that is inherited from bacteria. AB - Fe/S clusters are co-factors of numerous proteins with important functions in metabolism, electron transport and regulation of gene expression. Presumably, Fe/S proteins have occurred early in evolution and are present in cells of virtually all species. Biosynthesis of these proteins is a complex process involving numerous components. In mitochondria, this process is accomplished by the so-called ISC (iron-sulfur cluster assembly) machinery which is derived from the bacterial ancestor of the organelles and is conserved from lower to higher eukaryotes. The mitochondrial ISC machinery is responsible for biogenesis iron sulfur proteins both within and outside the organelle. Maturation of the latter proteins involves the ABC transporter Atm1p which presumably exports iron-sulfur clusters from the organelle. This review summarizes recent developments in our understanding of the biogenesis of iron-sulfur proteins both within bacteria and eukaryotes. PMID- 11004454 TI - How do uncoupling proteins uncouple? AB - According to the proton buffering model, introduced by Klingenberg, UCP1 conducts protons through a hydrophilic pathway lined with fatty acid head groups that buffer the protons as they move across the membrane. According to the fatty acid protonophore model, introduced by Garlid, UCPs do not conduct protons at all. Rather, like all members of this gene family, they are anion carriers. A variety of anions are transported, but the physiological substrates are fatty acid (FA) anions. Because the carboxylate head group is translocated by UCP, and because the protonated FA rapidly diffuses across the membrane, this mechanism permits FA to behave as regulated cycling protonophores. Favoring the latter mechanism is the fact that the head group of long-chain alkylsulfonates, strong acid analogues of FA, is also translocated by UCP. PMID- 11004455 TI - Effects of nitric oxide and peroxynitrite on the cytochrome oxidase K(m) for oxygen: implications for mitochondrial pathology. AB - This review summarises current knowledge about the effect of oxygen on cytochrome oxidase activity in vitro and in vivo. Cytochrome oxidase normally operates above its K(m) for oxygen in vivo. However, decreases in the intracellular oxygen concentration (hypoxia) under physiological extremes, or during pathophysiology, can cause mitochondrial respiration to become oxygen limited. Inhibitors that raise the enzyme's K(m) will induce oxygen limitation under apparently normoxic conditions. It is known that the concentrations of nitric oxide and peroxynitrite are raised in a number of pathophysiological conditions. These compounds are capable of reversibly and irreversibly raising the cytochrome oxidase K(m) for oxygen. Therefore, measurements of cell and mitochondrial respiration in vitro that fail to systematically vary oxygen through the range of physiological concentrations are likely to underestimate the effects of nitric oxide and peroxynitrite in vivo. PMID- 11004456 TI - Mitochondrial bioenergetics in aging. AB - Mitochondria are strongly involved in the production of reactive oxygen species, considered as the pathogenic agent of many diseases and of aging. The mitochondrial theory of aging considers somatic mutations of mitochondrial DNA induced by oxygen radicals as the primary cause of energy decline; experimentally, complex I appears to be mostly affected and to become strongly rate limiting for electron transfer. Mitochondrial bioenergetics is also deranged in human platelets upon aging, as shown by the decreased Pasteur effect (enhancement of lactate production by respiratory chain inhibition). Cells counteract oxidative stress by antioxidants; among lipophilic antioxidants, coenzyme Q is the only one of endogenous biosynthesis. Exogenous coenzyme Q, however, protects cells from oxidative stress by conversion into its reduced antioxidant form by cellular reductases. PMID- 11004457 TI - Reversal of nitric oxide-, peroxynitrite- and S-nitrosothiol-induced inhibition of mitochondrial respiration or complex I activity by light and thiols. AB - Nitric oxide (NO) and its derivatives peroxynitrite and S-nitrosothiols inhibit mitochondrial respiration by various means, but the mechanisms and/or the reversibility of such inhibitions are not clear. We find that the NO-induced inhibition of respiration in isolated mitochondria due to inhibition of cytochrome oxidase is acutely reversible by light. Light also acutely reversed the inhibition of respiration within iNOS-expressing macrophages, and this reversal was partly due to light-induced breakdown of NO, and partly due to reversal of the NO-induced inhibition of cytochrome oxidase. NO did not cause inhibition of complex I activity within isolated mitochondria, but 0.34 mM peroxynitrite, 1 mM S-nitroso-N-acetylpenicillamine or 1 mM S-nitrosoglutathione did cause substantial inhibition of complex I activity. Inhibition by these reagents was reversed by light, dithiothreitol or glutathione-ethyl ester, either partially or completely, depending on the reagent used. The rapid inhibition of complex I activity by S-nitroso-N-acetylpenicillamine also occurred in conditions where there was little or no release of free NO, suggesting that the inhibition was due to transnitrosylation of the complex. These findings have implications for the physiological and pathological regulation of respiration by NO and its derivatives. PMID- 11004458 TI - Re-emerging structures: continuing crystallography of the bacterial reaction centre. AB - The reaction centre is nature's solar battery, and is found in a number of variations on a common theme in plants, algae and photosynthetic bacteria. During the last 20 years, a combination of X-ray crystallography, spectroscopy and mutagenesis has provided increasingly detailed insights into the mechanism of light energy transduction in the bacterial reaction centre. This mini-review looks at the application of X-ray crystallography to the bacterial reaction centre, focussing in particular on recent information on the structural consequences of site-directed mutagenesis, the roles played by water molecules in the reaction centre, the mechanism of ubiquinone reduction, and studies of the phospholipid environment of the protein. PMID- 11004459 TI - Succinate: quinone oxidoreductases: new insights from X-ray crystal structures. AB - Membrane-bound succinate dehydrogenases (succinate:quinone reductases, SQR) and fumarate reductases (quinol:fumarate reductases, QFR) couple the oxidation of succinate to fumarate to the reduction of quinone to quinol and also catalyse the reverse reaction. SQR (respiratory complex II) is involved in aerobic metabolism as part of the citric acid cycle and of the aerobic respiratory chain. QFR is involved in anaerobic respiration with fumarate as the terminal electron acceptor, and is part of an electron transport chain catalysing the oxidation of various donor substrates by fumarate. QFR and SQR complexes are collectively referred to as succinate:quinone oxidoreductases (EC 1.3.5.1), have very similar compositions and are predicted to share similar structures. The complexes consist of two hydrophilic and one or two hydrophobic, membrane-integrated subunits. The larger hydrophilic subunit A carries covalently bound flavin adenine dinucleotide and subunit B contains three iron-sulphur centres. QFR of Wolinella succinogenes and SQR of Bacillus subtilis contain only one hydrophobic subunit (C) with two haem b groups. In contrast, SQR and QFR of Escherichia coli contain two hydrophobic subunits (C and D) which bind either one (SQR) or no haem b group (QFR). The structure of W. succinogenes QFR has been determined at 2.2 A resolution by X-ray crystallography (C.R.D. Lancaster, A. Kroger, M. Auer, H. Michel, Nature 402 (1999) 377-385). Based on this structure of the three protein subunits and the arrangement of the six prosthetic groups, a pathway of electron transfer from the quinol-oxidising dihaem cytochrome b to the site of fumarate reduction and a mechanism of fumarate reduction was proposed. The W. succinogenes QFR structure is different from that of the haem-less QFR of E. coli, described at 3.3 A resolution (T.M. Iverson, C. Luna-Chavez, G. Cecchini, D.C. Rees, Science 284 (1999) 1961-1966), mainly with respect to the structure of the membrane-embedded subunits and the relative orientations of soluble and membrane embedded subunits. Also, similarities and differences between QFR transmembrane helix IV and transmembrane helix F of bacteriorhodopsin and their implications are discussed. PMID- 11004460 TI - The mitochondrial cyanide-resistant oxidase: structural conservation amid regulatory diversity. AB - Mitochondria from all plants, many fungi and some protozoa contain a cyanide resistant, alternative oxidase that functions in parallel with cytochrome c oxidase as the terminal oxidase on the electron transfer chain. Characterization of the structural and potential regulatory features of the alternative oxidase has advanced considerably in recent years. The active site is proposed to contain a di-iron center belonging to the ribonucleotide reductase R2 family and modeling of a four-helix bundle to accommodate this active site within the C-terminal two thirds of the protein has been carried out. The structural features of this active site are conserved among all known alternative oxidases. The post translational regulatory features of the alternative oxidase are more variable among organisms. The plant oxidase is dimeric and can be stimulated by either alpha-keto acids or succinate, depending upon the presence or absence, respectively, of a critical cysteine residue found in a conserved block of amino acids in the N-terminal region of the plant protein. The fungal and protozoan alternative oxidases generally exist as monomers and are not subject to organic acid stimulation but can be stimulated by purine nucleotides. The origins of these diverse regulatory features remain unknown but are correlated with sequence differences in the N-terminal third of the protein. PMID- 11004461 TI - Crystallographic location of two Zn(2+)-binding sites in the avian cytochrome bc(1) complex. AB - The chicken mitochondrial ubiquinol cytochrome c oxidoreductase (bc(1) complex) is inhibited by Zn(2+) ions, but with higher K(i) ( approximately 3 microM) than the corresponding bovine enzyme. When equilibrated with mother liquor containing 200 microM ZnCl(2) for 7 days, the crystalline chicken bc(1) complex specifically binds Zn(2+) at 4 sites representing two sites on each monomer in the dimer. These two sites are close to the stigmatellin-binding site, taken to be center Q(o) of the Q-cycle mechanism, and are candidates for the inhibitory site. One binding site is actually in the hydrophobic channel between the Q(o) site and the bulk lipid phase, and may interfere with quinone binding. The other is in a hydrophilic area between cytochromes b and c(1), and might interfere with the egress of protons from the Q(o) site to the intermembrane aqueous medium. No zinc was bound near the putative proteolytic active site of subunits 1 and 2 (homologous to mitochondrial processing peptidase) under these conditions. PMID- 11004462 TI - Fusion protein approach to improve the crystal quality of cytochrome bo(3) ubiquinol oxidase from Escherichia coli. AB - Crystals of cytochrome bo(3) ubiquinol oxidase from E. coli diffract X-rays to 3.5 A and the structure determination is in progress. The limiting factor to the elucidation of the structural detail is the quality of the crystals; the diffraction spots from the crystals are diffused which leads to difficulties in processing the data beyond 4.0 A. Weak protein-protein contacts within the crystal lattice is assumed to be the cause of this problem. To improve these contacts, we have introduced protein Z to the C-terminal end of the subunit IV of cytochrome bo(3) and expressed both proteins as a single fusion. We have successfully obtained crystals of this fusion protein. The spot shape problem has clearly been solved in the crystals of the fusion protein although further optimization is necessary to obtain higher resolution. We also discuss the potential applications of this approach to the crystallization of membrane proteins in general. PMID- 11004463 TI - Proton-coupled electron transfer at the Q(o) site: what type of mechanism can account for the high activation barrier? AB - In Rhodobacter sphaeroides, transfer of the first electron in quinol oxidation by the bc(1) complex shows kinetic features (a slow rate (approx. 1.5 x 10(3)/s), high activation energy (approx. 65 kJ/mol) and reorganization energy, lambda (2.5 V)) that are unexpected from Marcus theory and the distances shown by the structures. Reduction of the oxidized iron-sulfur protein occurs after formation of the enzyme-substrate complex, and involves a H-transfer in which the electron transfer occurs through the approx. 7 A of a bridging histidine forming a H-bond with quinol and a ligand to 2Fe-2S. The anomalous kinetic features can be explained by a mechanism in which the electron transfer is constrained by coupled transfer of the proton. We discuss this in the context of mutant strains with modified E(m,7) and pK for the iron-sulfur protein, and Marcus theory for proton coupled electron transfer. We suggest that transfer of the second proton and electron involve movement of semiquinone in the Q(o) site, and rotation of the Glu of the conserved -PEWY- sequence. Mutational studies show a key role for the domain proximal to heme b(L). The effects of mutation at Tyr-302 (Tyr-279 in bovine sequence) point to a possible linkage between conformational changes in the proximal domain, and changes leading to closure of the iron-sulfur protein access channel at the distal domain. PMID- 11004464 TI - The cytochrome b(6)f complex: structural studies and comparison with the bc(1) complex. AB - Electron crystallography of the chloroplastic b(6)f complex allowed the calculation of projection maps of crystals negatively stained or embedded in glucose. This gives insights into the overall structure of the extra- and transmembrane domains of the complex. A comparison with the structure of the bc(1) complex, the mitochondrial homologue of the b(6)f complex, suggests that the transmembrane domains of the two complexes are very similar, confirming the structural homology deduced from sequence analysis. On the other hand, the extramembrane organisation of the c-type cytochrome and of the Rieske protein seems quite different. Nevertheless, the same type of movement of the Rieske protein is observed in the b(6)f as in the bc(1) complex upon the binding of the quinol analogue stigmatellin. Crystallographic data also suggest movements in the transmembrane domains of the b(6)f complex, which would be specific of the b(6)f complex. PMID- 11004465 TI - Protonation reactions in relation to the coupling mechanism of bovine cytochrome c oxidase. AB - Identification of the locations of protonatable sites in cytochrome c oxidase that are influenced by reactions in the binuclear centre is critical to assessment of proposed coupling mechanisms, and to controversies on where the pumping steps occur. One such protonation site is that which governs interconversion of the isoelectronic 607 nm 'P(M)' and 580 nm 'F' forms of the two-electron-reduced oxygen intermediate. Low pH favours protonation of a site that is close to an electron paramagnetic resonance (EPR)-silent radical species in P(M), and this induces a partial electronic redistribution to form an EPR detectable tryptophan radical in F. A further protonatable group that must be close to the binuclear centre has been detected in bacterial oxidases by Fourier transform infrared spectroscopy from pH-dependent changes in the haem-bound CO vibration frequency at low temperatures. However, in bovine cytochrome c oxidase under similar conditions of measurement, haem-bound CO remains predominantly in a single 1963 cm(-1) form between pH 6.5 and 8.5, indicating that this group is not present. Lack of pH dependence extends to the protein region of the CO photolysis spectra and suggests that both the reduced and the reduced/CO states do not have titratable groups that affect the binuclear centre strongly in the pH range 6.5 8.5. This includes the conserved glutamic acid residue E242 whose pK appears to be above 8.5 even in the fully oxidised enzyme. The results are discussed in relation to recent ideas on coupling mechanism. PMID- 11004466 TI - Towards understanding the chemistry of photosynthetic oxygen evolution: dynamic structural changes, redox states and substrate water binding of the Mn cluster in photosystem II. AB - This mini-review summarizes my postdoctoral research in the labs of T. Wydrzynski/C.B. Osmond, J.H.A. Nugent/M.C.W. Evans and V.K. Yachandra/K. Sauer/M.P. Klein. The results are reported in the context of selected data from the literature. Special emphasis is given to the mode of substrate water binding, Mn oxidation states and the structures of the Mn cluster in the four (meta)stable redox states of the oxygen evolving complex. The paper concludes with a working model for the mechanism of photosynthetic water oxidation that combines mu-oxo bridge oxidation in the S(3) state (V.K. Yachandra, K. Sauer, M.P. Klein, Chem. Rev. 96 (1996) 2927-2950) with O-O bond formation between two terminal Mn-hydroxo ligands during the S(3)-->(S(4))-->S(0) transition. PMID- 11004467 TI - Cloning and expression of cDNAs encoding plant V-ATPase subunits in the corresponding yeast null mutants. AB - Complementation of yeast null mutants is widely used for cloning of homologous genes from heterologous sources. We have used this method to clone the relevant V ATPase genes from lemon fruit and Arabidopsis thaliana cDNA libraries. The pH levels are very different in the vacuoles of the lemon fruit and the A. thaliana, yet both are the result of the activity of the same enzyme complex, namely the V ATPase. In order to investigate the mechanism that enables the enzyme to maintain such differences in pH values, we have compared the subunit composition of the V ATPase complex from both sources. Towards this end, we have constructed a cDNA library from lemon fruit and cloned it into a similar shuttle vector to the one of the A. thaliana cDNA library, which is commercially available. In this work, we report the cloning and expression of VMA10 from both sources, two isoforms of the lemon proteolipid (VMA3) and the lemon homologue of yeast VPH1/STV1 subunit, LEMAC. PMID- 11004468 TI - Biological nano motor, ATP synthase F(o)F(1): from catalysis to gammaepsilonc(10 12) subunit assembly rotation. AB - Proton translocating ATPase (ATP synthase), a chemiosmotic enzyme, synthesizes ATP from ADP and phosphate coupling with the electrochemical ion gradient across the membrane. This enzyme has been studied extensively by combined genetic, biochemical and biophysical approaches. Such studies revealed a unique mechanism which transforms an electrochemical ion gradient into chemical energy through the rotation of a subunit assembly. Thus, this enzyme can be defined as a nano motor capable of coupling a chemical reaction and ion translocation, or more simply, as a protein complex carrying out rotational catalysis. In this article, we briefly discuss our recent work, emphasizing the rotation of subunit assembly (gammaepsilonc(10-12)) which is formed from peripheral and intrinsic membrane subunits. PMID- 11004469 TI - Critical evaluation of the one- versus the two-channel model for the operation of the ATP synthase's F(o) motor. AB - The mechanism of converting an electrochemical gradient of protons or Na(+) ions across the membrane into rotational torque by the F(o) motor of the ATP synthase has been described by a two-channel model or by a one-channel model. Experimental evidence obtained with the F(o) motor from the Propionigenium modestum ATP synthase is described which is in accordance with the one-channel model, but not with the two-channel model. This evidence includes the ATP-dependent occlusion of one (22)Na(+) per ATP synthase with a mutated Na(+)-impermeable a subunit or the Na(+)(in)/(22)Na(+)(out) exchange which is not affected by modifying part of the c subunit sites with dicyclohexylcarbodiimide. PMID- 11004470 TI - The role of the D- and K-pathways of proton transfer in the function of the haem copper oxidases. AB - The X-ray structures of several haem-copper oxidases now at hand have given important constraints on how these enzymes function. Yet, dynamic data are required to elucidate the mechanisms of electron and proton transfer, the activation of O(2) and its reduction to water, as well as the still enigmatic mechanism by which these enzymes couple the redox reaction to proton translocation. Here, some recent observations will be briefly reviewed with special emphasis on the functioning of the so-called D- and K-pathways of proton transfer. It turns out that only one of the eight protons taken up by the enzyme during its catalytic cycle is transferred via the K-pathway. The D-pathway is probably responsible for the transfer of all other protons, including the four that are pumped across the membrane. The unique K-pathway proton may be specifically required to aid O-O bond scission by the haem-copper oxidases. PMID- 11004471 TI - A re-examination of the structural and functional consequences of mutation of alanine-128 of the b subunit of Escherichia coli ATP synthase to aspartic acid. AB - The effects of mutation of residue Ala-128 of the b subunit of Escherichia coli ATP synthase to aspartate on the structure of the subunit and its interaction with the F(1) sector were analyzed. Determination of solution molecular weights by sedimentation equilibrium ultracentrifugation revealed that the A128D mutation had little effect on dimerization in the soluble b construct, b(34-156). However, the mutation caused a structural perturbation detected through both a 12% reduction in the sedimentation coefficient and also a reduced tendency to form intersubunit disulfide bonds between cysteine residues inserted at position 132. Unlike the wild-type sequence, the A128D mutant was unable to interact with F(1) ATPase. These results indicate that the A128D mutation caused a structural change in the C-terminal region of the protein, preventing the binding to F(1) but having little or no effect on the dimeric nature of b. PMID- 11004472 TI - Light-induced FTIR difference spectroscopy of the S(2)-to-S(3) state transition of the oxygen-evolving complex in Photosystem II. AB - We have applied flash-induced FTIR spectroscopy to study structural changes upon the S(2)-to-S(3) state transition of the oxygen-evolving complex (OEC) in Photosystem II (PSII). We found that several modes in the difference IR spectrum are associated with bond rearrangements induced by the second laser flash. Most of these IR modes are absent in spectra of S(2)/S(1), of the acceptor-side non heme ion, of Yradical(D)/Y(D) and of S(3)'/S(2)' from Ca-depleted PSII preparations. Our results suggest that these IR modes most likely originate from structural changes in the oxygen-evolving complex itself upon the S(2)-to-S(3) state transition in PSII. PMID- 11004473 TI - Proton transfer from glutamate 286 determines the transition rates between oxygen intermediates in cytochrome c oxidase. AB - We have investigated the electron-proton coupling during the peroxy (P(R)) to oxo ferryl (F) and F to oxidised (O) transitions in cytochrome c oxidase from Rhodobacter sphaeroides. The kinetics of these reactions were investigated in two different mutant enzymes: (1) ED(I-286), in which one of the key residues in the D-pathway, E(I-286), was replaced by an aspartate which has a shorter side chain than that of the glutamate and, (2) ML(II-263), in which the redox potential of Cu(A) is increased by approximately 100 mV, which slows electron transfer to the binuclear centre during the F-->O transition by a factor of approximately 200. In ED(I-286) proton uptake during P(R)-->F was slowed by a factor of approximately 5, which indicates that E(I-286) is the proton donor to P(R). In addition, in the mutant enzyme the F-->O transition rate displayed a deuterium isotope effect of approximately 2.5 as compared with approximately 7 in the wild-type enzyme. Since the entire deuterium isotope effect was shown to be associated with a single proton-transfer reaction in which the proton donor and acceptor must approach each other (M. Karpefors, P. Adelroth, P. Brzezinski, Biochemistry 39 (2000) 6850), the smaller deuterium isotope effect in ED(I-286) indicates that proton transfer from E(I-286) determines the rate also of the F-->O transition. In ML(II 263) the electron-transfer to the binuclear centre is slower than the intrinsic proton-transfer rate through the D-pathway. Nevertheless, both electron and proton transfer to the binuclear centre displayed a deuterium isotope effect of approximately 8, i.e., about the same as in the wild-type enzyme, which shows that these reactions are intimately coupled. PMID- 11004474 TI - Steady-state tyrosine fluorescence to study the lipid-binding properties of a wheat non-specific lipid-transfer protein (nsLTP1). AB - The binding properties of a wheat non-specific lipid-transfer protein (nsLTP1) for different mono- and diacylated lipids was investigated. Lipids varied by their chain length, unsaturation and/or polar head group. In the case of fatty acid or lysophospholipid with a C10 chain length, no interaction can be measured, while poor affinity is reported for a C12 chain length. The dissociation constant (K(d)) is about 0.5 uM independent of chain length from C14 to C18. The same affinity is obtained for C18 fatty acids with one or two unsaturations, whatever the cis-trans double bond isomery. In all cases, the number of binding sites, n, by protein ranges between 1.6 and 1.9, suggesting that two lipids can fit within the protein. omega-Hydroxy-palmitic acid, a natural monomer of cutin polymer, is found to interact with nsLTP1 with a K(d) of 1 uM and n=2. In contrast with previous data that reported the binding of the anionic diacylated phospholipid, DMPG (Sodano et al., FEBS Lett. 416 (1997) 130-134), nsLTP1 is not able to bind dimyristoylphosphatidylcholine, dimyristoylphosphatidic acid, palmitoyl oleoylphosphatidylcholine or palmitoyl-oleoylphosphatidylglycerol added as liposomes or solubilized in ethanol. However, when both nsLTP1 and lipids are first solubilized in methanol, and then in the buffer, it was evidenced that the protein can bind these lipids. These results suggest that lipid-lipid interactions play an essential role in the binding process of plant nsLTP1 as previously mentioned for other lipid-transfer proteins. PMID- 11004475 TI - Plasma membrane coating with cationic silica particles and osmotic shock alters the morphology of bovine aortic endothelial cells. AB - We have used a published method of membrane preparation based on the precoating of the apical membrane of aortic endothelial cells with cationic silica microbeads (with or without polyacrylic acid) in combination with an osmotic shock and mechanical shearing to isolate the apical from the basal plasma membranes of these cells, in vitro. After labeling of the plasma membrane of adherent endothelial cells with a fluorescent derivative of phosphatidylcholine and by using laser confocal fluorescence scanning microscopy, we found that this method of membrane isolation rapidly induced invaginations of the basal plasma membrane to an extent which makes this method unsuitable for further membrane lipid analysis. Morphological analysis of the cells and fluorescence recovery after photobleaching experiments on the plasma membranes were performed at each step of the purification procedure and showed that only hypotonic shock and mechanical shearing of the cells enabled the basal plasma membranes to be purified without significant morphological changes. PMID- 11004477 TI - The novel anti-tumour agent oxamflatin differentially regulates urokinase and plasminogen activator inhibitor type 2 expression and inhibits urokinase-mediated proteolytic activity. AB - Cell surface, urokinase (u-PA)-mediated, plasminogen activation has recently been recognised as a process integral to extracellular matrix degradation. The primary inhibitor of u-PA activity in the extracellular matrix is plasminogen activator inhibitor type 2 (PAI-2), a serine protease inhibitor. The malignant metastatic phenotype is associated with excessive and uncontrolled, tumour cell-associated, u-PA-mediated, extracellular matrix degradation. Inhibition of the malignant metastatic phenotype via induction of PAI-2 expression and/or inhibition of u-PA expression may represent a novel means via which the metastatic phenotype can be arrested. Agents capable of inducing PAI-2 and/or inhibiting u-PA activity may restrict u-PA-mediated tumour cell proteolysis and facilitate in the development of therapeutic strategies to combat malignant disease. We have identified the hydroxamic acid derivative oxamflatin, previously noted to revert the malignant phenotype in K-ras-transformed NIH-3T3 cells, as capable of upregulating PAI-2 and simultaneously suppressing u-PA expression in two different cell systems. In addition, zymographic analysis indicated that oxamflatin treatment results in a significant reduction in u-PA proteolytic activity in both HT-1080 fibrosarcoma and U-937 histiocytic lymphoma cells. We postulate that oxamflatin represents a novel means by which induction of PAI-2 and concomitant inhibition of u-PA gene and protein expression can be achieved and may be of benefit in inhibiting the malignant metastatic phenotype. PMID- 11004478 TI - Isolation and characterization of a novel liver-specific gene, hepassocin, upregulated during liver regeneration. AB - By differential cDNA cloning coupled with Xenopus oocyte expression screening, we isolated a cDNA encoding a novel protein, termed 'hepassocin', the expression of which is upregulated in the regenerating rat liver. The cDNA contained a single open reading frame encoding a protein of 314 amino acids (ca. 34 kDa), including 24 amino acids of signal sequence. The protein expressed from the cDNA in Verots cells had activity to stimulate DNA synthesis in primary rat hepatocytes and was of 66 kDa or 34 kDa, under non-reducing or reducing conditions, respectively. Using an affinity column conjugated with the antibody raised against a peptide in a hydrophilic region, we purified hepassocin from the rat liver: it had a DNA synthesis-stimulating activity in hepatocytes. The hepassocin obtained here was 66 kDa, and the 34 kDa protein obtained under reducing conditions contained five cysteine residues, indicating that hepassocin is active as a homodimer. Northern blot analysis revealed that hepassocin mRNA (1.4 kb in length) occurred only in the liver, and in situ hybridization studies revealed its presence in parenchymal hepatocytes but not in endothelial cells. Furthermore, the expression of hepassocin mRNA was upregulated during compensatory hyperplasia after partial hepatectomy and regeneration after galactosamine treatment in the rat liver. These results suggest that hepassocin plays an important role in stimulating liver cell growth, through an autocrine mechanism. PMID- 11004479 TI - Transforming growth factor-beta(1) regulation of surfactant protein B gene expression is mediated by protein kinase-dependent intracellular translocation of thyroid transcription factor-1 and hepatocyte nuclear factor 3. AB - The transforming growth factor-beta (TGF beta) polypeptides control a variety of cellular processes including organogenesis and cellular proliferation and differentiation. In the developing lung, TGF beta(1) treatment inhibits airway branching and expression of the genes for surfactant proteins (SP). Many effects of TGF beta are mediated at the level of gene transcription but there is limited information regarding signaling pathways and target transcription factors. In this study with human pulmonary adenocarcinoma H441 cells, we investigated TGF beta(1) effects on SP-B, a protein which is essential for normal function of pulmonary surfactant. TGF beta(1) (10 ng/ml) reduced SP-B mRNA content in a time dependent fashion, and transient transfection studies localized responsiveness to the region of the SP-B promoter (-112/-72 bp) containing binding sites for thyroid transcription factor-1 (TTF-1) and hepatocyte nuclear factor 3 (HNF3), transcription factors that are important enhancers of SP gene expression. Using electrophoretic mobility shift assay and immunofluorescence, we demonstrated rapid accumulation of these transcription factors in the cytoplasm and subsequent loss from the nucleus on TGF beta(1) treatment of both adenocarcinoma cells and cultured human fetal lung. TGF beta(1) treatment caused intracellular translocation of protein kinase C and effects of TGF beta(1) were mostly abrogated in the presence of the protein kinase inhibitor calphostin C. We conclude that TGF beta(1), acting via protein phosphorylation, blocks nuclear translocation of TTF-1 and HNF3 which results in down-regulation of the SP-B gene and presumably other pulmonary genes which are transactivated by these factors. PMID- 11004480 TI - Localization, expression and genomic structure of the gene encoding the human serine protease testisin. AB - Testisin is a recently identified human serine protease expressed by premeiotic testicular germ cells and is a candidate tumor suppressor for testicular cancer. Here, we report the characterization of the gene encoding testisin, designated PRSS21, and its localization on the short arm of human chromosome 16 (16p13.3) between the microsatellite marker D16S246 and the radiation hybrid breakpoint CY23HA. We have further refined the localization to cosmid 406D6 in this interval and have established that the gene is approximately 4. 5 kb in length, and contains six exons and five intervening introns. The structure of PRSS21 is very similar to the human prostasin gene (PRSS8) which maps nearby on 16p11.2, suggesting that these genes may have evolved through gene duplication. Sequence analysis showed that the two known isoforms of testisin are generated by alternative pre-mRNA splicing. A major transcription initiation site was identified 97 nucleotides upstream of the testisin translation start and conforms to a consensus initiator element. The region surrounding the transcription initiation site lacks a TATA consensus sequence, but contains a CCAAT sequence and includes a CpG island. The 5'-flanking region contains several consensus response elements including Sp1, AP1 and several testis-specific elements. Analysis of testisin gene expression in tumor cell lines shows that testisin is not expressed in testicular tumor cells but is aberrantly expressed in some tumor cell lines of non-testis origin. These data provide the basis for identifying potential genetic alterations of PRSS21 that may underlie both testicular abnormalities and tumorigenesis. PMID- 11004481 TI - Distribution of mRNA encoding B78/apj, the rat homologue of the human APJ receptor, and its endogenous ligand apelin in brain and peripheral tissues. AB - The human APJ receptor is a G protein-coupled receptor which functions as an efficient alternative co-receptor for a number of human immunodeficiency virus type 1 and simian immunodeficiency virus strains. We have cloned the rat APJ receptor, which we term B78/apj, and have mapped the mRNA distribution of both the receptor and its natural ligand apelin in rat tissues. Northern blot analysis showed a similar pattern of expression for B78/apj and apelin mRNAs with hybridising transcripts seen in the lung, heart, skeletal muscle, kidney, brain and liver. In situ hybridisation histochemistry studies revealed intense B78/apj gene expression in the parenchyma of the lung, a sub-population of glomeruli in the kidney, the corpora lutea of the ovary and isolated cells of the anterior lobe of the pituitary. B78/apj mRNA had a striking and unique distribution within the central nervous system (CNS) where receptor expression was found in cells within the meninges around the brain, in the posterior magnocellular and medial parvocellular areas of the hypothalamic paraventricular nucleus and in the supraoptic nucleus. This hypothalamic distribution offers a possible specific role of this receptor in mediating neuroendocrine responses in the CNS. PMID- 11004482 TI - Cloning of cDNAs encoding retinoic acid receptors RAR gamma 1, RAR gamma 2, and a new splicing variant, RAR gamma 3, from Aambystoma mexicanum and characterization of their expression during early development. AB - To analyze retinoic acid (RA) receptor (RAR) expression during early development in the urodele embryo, we have isolated cDNAs for four members of the axolotl (Ambystoma mexicanum) RAR family, namely RAR alpha (NR1B1), aRAR gamma 1 (NR1B3a), aRAR gamma 2 (NR1B3b), and a new splicing variant of aRAR gamma 2, aRAR gamma 3 (NR1B3c), which contains an insertion of five hydrophobic amino acids in the C-terminal region of the DNA binding domain. The temporal expression pattern of the RAR gamma isoforms was established by RT-PCR using total RNA from embryos of different stages. The expression of aRAR gamma 2 coincides with neurulation and is enhanced in the extremities of the embryo's anteroposterior axis. The aRAR gamma 3 is specifically expressed during gastrulation and early neurulation, whereas aRAR gamma 1 is expressed later during organogenesis. Global aRAR gamma 2 mRNA levels, as well as their spatio-temporal expression pattern in the neurula, were not affected by treatment with RA. These results show that several RARs are expressed in the axolotl embryo during early development, and reveal the existence of a new RAR gamma variant. PMID- 11004483 TI - Characterization of two putative histone deacetylase genes from Aspergillus nidulans. AB - In eukaryotic organisms, acetylation of core histones plays a key role in the regulation of transcription. Multiple histone acetyltransferases (HATs) and histone deacetylases (HDACs) maintain a dynamic equilibrium of histone acetylation. The latter form a highly conserved protein family in many eukaryotic species. In this paper, we report the cloning and sequencing of two putative histone deacetylase genes (rpdA, hosA) of Aspergillus nidulans, which are the first to be analyzed from filamentous fungi. Hybridization with a chromosome specific cosmid library of A. nidulans allowed the localization of rpdA to chromosome III and hosA to chromosome II, respectively. PCR analyses and Southern hybridization experiments revealed that no further members of the RPD3 family are present in the genome of the fungus. Although sequence alignment displays significant amino acid similarity to other eukaryotic RPD3-type deacetylases, the deduced RPDA sequence reveals an unusual 200-amino acid extension at the C terminus. Expression of both genes was determined by RNA blot analysis. Treatment of the cells with trichostatin A (TSA), a potent inhibitor of HDACs, was found to stimulate expression of rpdA of A. nidulans. PMID- 11004484 TI - Expression of purinergic receptors (ionotropic P2X1-7 and metabotropic P2Y1-11) during myeloid differentiation of HL60 cells. AB - The expression of human purinergic P2 receptors (P2X1-7 and P2Y1-11) as well as the ecto-enzymes apyrase (CD39) and 5'-nucleotidase (CD73) was investigated on the nucleic acid level during granulocytic and monocytic differentiation of HL60 cells and on peripheral human blood leukocytes. RT-PCR and dot-blot hybridization assays indicated that mRNA transcripts of all analyzed P2 receptors apart from the P2X3 receptor were expressed during myeloid development of HL60 cells, showing a distinct regulation during the course of differentiation. In blood leukocytes, transcripts of P2X5, P2X7 and all P2Y receptors, except for P2Y6, receptor were found. CD39 and CD73 showed a marked upregulation during myeloid maturation. Functional analysis of P2 receptor-mediated intracellular Ca(2+) increase after stimulation with ATP revealed no change during granulocytic differentiation, but showed a strong attenuation in both potency and efficacy during monocytic development of HL60 cells. PMID- 11004485 TI - cDNA structure, genomic organization, and promoter analysis of the mouse intestinal peptide transporter PEPT1. AB - We describe in this report the cDNA structure, functional characteristics, genomic organization, and promoter analysis of the mouse H(+)-coupled low affinity peptide transporter PEPT1. The mouse PEPT1 cDNA cloned from a kidney cDNA library is approximately 3.1 kb long and encodes a protein of 709 amino acids. When expressed heterologously in mammalian cells and in Xenopus laevis oocytes, mouse PEPT1 mediates H(+)-coupled electrogenic transport of the dipeptide glycylsarcosine. The mouse pept1 gene, cloned from a genomic DNA library in bacterial artificial chromosome, is approximately 38 kb long and consists of 23 exons and 22 introns. 5'-Rapid amplification of cDNA ends with poly(A)(+) RNA from mouse intestine has identified the transcription start site that lies 31 bp upstream of the translation start site. The promoter region upstream of the transcription start site does not contain the TATA box but possesses three GC boxes which are the binding sites for the transcription activator SP1. Functional analysis of the promoter region using the luciferase reporter assay in Caco-2 cells (a human intestinal cell line that express PEPT1 constitutively) and five different 5'-deletion fragments of the promoter has shown that essential promoter/enhancer elements are present within 1140 bp upstream of the transcription start site. PMID- 11004487 TI - Analysis of the promoter region of the human FcRn gene. AB - The 5'-flanking region of the human FcRn alpha-chain gene was analyzed for its ability to directly express the chloramphenicol acetyltransferase (CAT) reporter gene in NIH3T3 and Lu106 cells. Transient transfection of the CAT constructs revealed that there was promoter activity in the region -660 to +300 of the 5' flanking sequence. Electrophoretic mobility-shift assays showed that there are functional binding sites for Sp1 or Sp1-like factors, AP1 or a related factor, and additional unidentified proteins in the promoter region. PMID- 11004486 TI - Interleukin-6 modulates interferon-regulated gene expression by inducing the ISGF3 gamma gene using CCAAT/enhancer binding protein-beta(C/EBP-beta). AB - Although interleukin-6 (IL-6) alone does not induce the expression of IFN stimulated genes (ISG), a low dose priming of cells with IL-6 strongly enhances the cellular responses to interferon-alpha (IFN-alpha). This effect of IL-6 is not due to superstimulation of the JAK-STAT pathway. Rather, IL-6 induces expression of ISGF3 gamma (p48), a subunit of the multimeric transcription factor ISGF3. As a result IFN-alpha robustly activates gene transcription in IL-6 primed cells. We have shown earlier that the transcription of ISGF3 gamma gene is regulated through a novel element GATE (gamma-IFN activated transcriptional element). We show here IL-6 induces the ISGF3 gamma gene through GATE. Transcription factor C/EBP-beta is required for inducing ISGF3 gamma gene expression through GATE. A mutant C/EBP-beta inhibits the IL-6 inducible ISGF3 gamma gene expression through GATE. Together, these results establish a molecular basis for the synergy between IFNs and IL-6. PMID- 11004488 TI - Structural organization and expression of the gaegurin 4 gene of Rana rugosa. AB - Gaegurin 4 (GGN4) is a member of the antimicrobial peptide subfamily isolated from the skin of Rana rugosa. We cloned gDNA encoding GGN4 to study its gene organization and regulation of expression. The GGN4 gene occurs in single copy in the R. rugosa genome and contains a single intron of about 3.4 kb. The transcription start site is located 68 bases upstream of the translation initiation codon. The GGN4 gene was expressed both in Xenopus kidney epithelial cells (A6) and in Xenopus oocytes using the chloramphenicol acetyltransferase reporter gene system. The 5' flanking region of the GGN4 gene contains a dl binding site that is known to regulate acute phase immune response related gene expression in mammals and insects. The dl protein bound specifically to the GGN4 gene promoter region. Mutants that serially delete the 5' flanking region show that removal of the dl binding site inhibited GGN4 gene expression in both A6 cells and Xenopus oocytes. From these results, we propose that expression of the GGN4 gene may be regulated by the region containing the dl element which plays a key role in the regulation of antimicrobial peptide genes in Drosophila and mammals. PMID- 11004489 TI - Cloning and characterization of a novel RNA-binding protein SRL300 with RS domains. AB - AT-rich element binding factor 1 (ATBF1) mRNA encodes a transcription factor implicated in neuronal differentiation. A cDNA for the protein that can bind the 5'-noncoding sequence of the ATBF1 mRNA was cloned. The deduced protein, termed SRL300, contains a unique RNA-binding region, two large RS domains and many phosphorylation sites. SRL300 protein was detected in both human and rat cells. PMID- 11004490 TI - Identification of a novel platelet-derived growth factor-like gene, fallotein, in the human reproductive tract. AB - We isolated the cDNA of a novel platelet-derived growth factor-like gene from human endometrium. The gene was named fallotein; it was 3007 bases in length, and encoded a protein of 345 amino acids. Antiserum against the fallotein protein can recognize a specific protein in the fallopian tube, with a molecular size in accordance with the anticipated size of fallotein. Fallotein mRNA is expressed in two molecular sizes, 3.8 and 2.9 kb, with the former being more abundant. High expression of the gene was found in the prostate, testis, and uterus. A weaker expression signal was found in the spleen, thymus, and small intestine, but expression of fallotein in the colon and peripheral blood leukocytes was negligible. PMID- 11004491 TI - Structure and stress-related expression of two cDNAs encoding proteinase inhibitor II of Nicotiana glutinosa L. AB - Two cDNAs, pNGPI-1 and pNGPI-2, encoding Nicotiana glutinosa proteinase inhibitor II (PI-II) have been cloned, sequenced and identified. The deduced amino acid sequences are 54-82% identical to those of other plant PI-II. The NGPI-1 protein is composed of eight repeated domains, while NGPI-2 contains six repeated regions, each with a putative reactive site. The expression of NGPI-1 is highly regulated in a developmental- and tissue-specific manner, with the transcript being detected in young leaves and floral organs of N. glutinosa plants. In mature leaves, the NGPI-1 gene is rapidly activated by distinct temporal induction patterns in response to pathogen-related (biotic) and wound-related (abiotic) stresses. PMID- 11004492 TI - Genomic organization and transcriptional regulation of maize ZmRR1 and ZmRR2 encoding cytokinin-inducible response regulators. AB - Maize genomic clones encoding cytokinin-inducible response regulators, ZmRR1 and ZmRR2, have been isolated. In comparison with the corresponding cDNAs, ZmRR2 was found to be interrupted in the translated region by an intron whereas ZmRR1 was not. The 5'-flanking regions of the two genes shared conserved regions and putative cis-elements, which had been identified in maize or other plant species. The run-on transcription assay and the analysis of stable maize transformants of ZmRR1 promoter-beta-glucuronidase fusion gene revealed that the accumulation of the transcripts in response to cytokinins is, at least in parts, attributed by transcriptional activation. PMID- 11004493 TI - Nucleotide sequence of cDNA clones coding for a brain-type fatty acid binding protein and its tissue-specific expression in adult zebrafish (Danio rerio). AB - We have determined the nucleotide sequence for two cDNA clones coding for a fatty acid binding protein (FABP) from zebrafish (Danio rerio). Comparison of the sequence with GenBank entries revealed extensive amino acid identity between this zebrafish FABP and brain FABPs (B-FABP) from other species. The zebrafish B-FABP cDNA hybridized to single restriction fragments of total zebrafish genomic DNA digested with the restriction endonucleases BglII or EcoRI suggesting that a single copy of the B-FABP gene is present in the zebrafish genome. Northern blot analysis demonstrated that the zebrafish B-FABP mRNA is approximately 850 nucleotides in length. In situ hybridization revealed that the B-FABP mRNA was expressed in the periventricular gray zone of the optic tectum of the adult zebrafish brain. PMID- 11004494 TI - cDNA sequence and tissue-specific expression of a basic liver-type fatty acid binding protein in adult zebrafish (Danio rerio). AB - We have determined the nucleotide sequence for a cDNA clone derived from zebrafish (Danio rerio) that codes for a fatty acid binding protein (FABP). Amino acid sequence similarity of the zebrafish FABP was highest to FABPs isolated from the livers of catfish, chicken, salamander and iguana. The open-reading frame of the zebrafish FABP cDNA codes for a protein of 14.0 kDa with a calculated isoelectric point of 8.8. The zebrafish liver-type FABP (L-FABP) cDNA hybridized to single restriction fragments of total zebrafish genomic DNA digested with the restriction endonucleases HaeIII or EcoRI suggesting that a single copy of the L FABP gene is present in the zebrafish genome. Northern blot analysis demonstrated that the zebrafish L-FABP cDNA hybridized to a transcript of 700 nucleotides in total zebrafish RNA. In situ hybridization and emulsion autoradiography revealed that the L-FABP was expressed exclusively in the liver of the adult zebrafish. Based on amino acid sequence similarity, the isoelectric point and its tissue specific pattern of expression, we conclude that this zebrafish FABP belongs to the basic liver-type FABPs only found, thus far, in non-mammalian vertebrates. PMID- 11004495 TI - Characterisation of a cysteine protease cDNA from Lolium multiflorum leaves and its expression during senescence and cytokinin treatment. AB - A cysteine protease cDNA clone (See1) highly homologous to barley aleurain was isolated from Lolium multiflorum leaves. During leaf senescence, expression of the See1 mRNA and protein was strongly enhanced. In dark-incubated leaf segments, cytokinin delayed senescence and reduced expression of both See1 mRNA and protein. PMID- 11004496 TI - New isoforms of cytochrome c oxidase subunit IV in tuna fish. AB - In the present study, the cDNA sequences of cytochrome c oxidase subunit IV isoforms from tuna fish are reported. The cDNAs share 57% identity among each other and the deduced amino acid sequences of the mature proteins 56% identity. Until now, only in yeast are two isoforms of the corresponding subunit V known, which are expressed in response to the oxygen supply. The hypothetical function of the new isoforms in fish for adaptation to different oxygen partial pressures in tissues of higher organisms is discussed. PMID- 11004497 TI - Molecular characterization of Drosophila melanogaster dihydropteridine reductase. AB - Dihydropteridine reductase (DHPR) catalyzes the NAD(P)H-mediated reduction of quinonoid dihydropteridine as a part of pterin-dependent aromatic amino acid hydroxylation. We isolated a fragment of Drosophila DHPR gene by PCR using degenerate primers. By screening a cDNA library, we obtained full-length clones. The predicted amino acid sequence of the Drosophila DHPR protein was highly homologous to other species including human and mouse. In particular, the Tyr (Xaa)(3)-Lys motif, known as the NAD(P)H binding domain, and most amino acids relevant to quinonoid dihydropteridine binding site are identical to human DHPR. The recombinant DHPR protein expressed in Escherichia coli showed DHPR enzyme activity. Northern blot analysis revealed two transcripts of 1.1 and 0.9 kb. Genomic DNA sequencing revealed that the gene consists of two exons interrupted by a single 96-bp intron. The two transcripts have alternative promoters, both having no putative TATA box or CAAT box, but sharing a common poly(A)(+) signal. The existence of two alternative promoters suggests that each transcript be regulated independently through different stimuli. Further study is needed to examine the expression and function of the two alternative transcripts. PMID- 11004498 TI - Isolation and sequence of the human cytochrome c oxidase subunit VIIaL gene. AB - The gene for human cytochrome c oxidase subunit VIIa liver isoform (COX7AL) was isolated and its sequence determined and analyzed. The three introns of the gene are considerably larger than those of the heart isoform of subunit VIIa (COX7AH), but the position of the introns relative to the cDNA sequences is homologous between the two genes. Comparison with other isolated COX7AL genes suggests that the promoter region binding motifs for transcription factors have evolved along with the coding region. In fibroblasts cultured originally from a Leigh's disease patient, a shortened COX7AL cDNA was identified by RT-PCR, consisting of exon I joined to exon IV, omitting exons II and III. No mutation could be identified in COX7AL of the patient, suggesting that the shortened cDNA is due to an alteration of the genome during cell culture. A surprising transcription of COX7AH was observed in cultured fibroblasts, suggesting a potential utility of these cells for study of its gene expression. PMID- 11004499 TI - Molecular cloning of cDNA encoding a ubiquitin-activating enzyme (E1) from goldfish (Carassius auratus) and expression analysis of the cloned gene. AB - Destruction of cyclin B is required to the mitotic and meiotic cycles. A cyclin specific ubiquitinating system, including ubiquitin-activating enzyme (E1), is thought to be responsible for cyclin B destruction. Here we present the cloning, sequencing and expression analysis of goldfish, Carassius auratus, E1 from goldfish ovary. The cloned cDNA is 4069 bp long and encodes 1059 amino acids. The deduced amino acid sequence is highly homologous to E1 from other species. Recombinant goldfish E1 could transfer ubiquitin to cyclin-selective ubiquitin conjugating enzyme. Tissue distribution revealed a single 4.0-kb message ubiquitous among tissues. PMID- 11004500 TI - Identification of a novel human tRNA(Ser(CGA)) functional in murine leukemia virus replication. AB - We have identified a human tRNA(Ser) isoacceptor matching the UCG codon. The tRNA was discovered via its ability to act in reverse transcription of a murine leukemia virus vector containing a complementary tRNA primer binding site (Lund et al., Nucleic Acids Res., 28 (2000) 791-799). The tRNA(Ser(CGA)) was detected in cell lines of human, monkey and mouse origin. The UCG codon is the most rarely used codon in human genes. The cloned human tRNA(Ser(CGA)) gene encodes an 85 nucleotide, intron-less tRNA, contains a consensus split intragenic promoter and is located at region p21.3-22.2 on chromosome 6. The integrity and functionality of the cloned tRNA(Ser(CGA)) gene was verified by in vitro transcription analysis in HeLa nuclear extracts. PMID- 11004501 TI - Cloning, sequencing and expression of the tetraheme cytochrome c(3) from Desulfovibrio gigas. AB - The gene encoding the tetraheme cytochrome c(3) from Desulfovibrio gigas was cloned and sequenced from a 2.7-kb EcoRI-PstI insert of D. gigas DNA. The derived amino acid sequence showed that the D. gigas cytochrome c(3) is synthesized as a precursor protein with an N-terminal signal peptide sequence of 25 residues and allowed the correction of the previous reported amino acid sequence (Matias et al. Protein Science 5 (1996) 1342-1354). Expression in D. vulgaris (Hildenborough) was possible by conjugal transfer of a recombinant broad-host range vector pSUP104 containing a SmaI fragment of the D. gigas cytochrome c(3) gene. Biochemical, immunological and spectroscopic analysis of the purified protein showed that the recombinant cytochrome is identical to that isolated from D. gigas. PMID- 11004502 TI - Molecular cloning and expression of rat betacellulin cDNA. AB - The cDNA encoding an entire open reading frame of rat betacellulin has been cloned from rat kidney. Expression of this cDNA in COS7 cells showed a significant amount of mitogenic activity in the culture media. Western blotting of the cell lysates suggested that the membrane-anchored precursor was cleaved to release its ectodomain very efficiently. PMID- 11004503 TI - Differential developmental expression and cell type specificity of Dictyostelium catalases and their response to oxidative stress and UV-light. AB - Cells of Dictyostelium discoideum are highly resistant to DNA damaging agents such as UV-light, gamma-radiation and chemicals. The genes encoding nucleotide excision repair (NER) and base excision repair (BER) enzymes are rapidly upregulated in response to UV-irradiation and DNA-damaging chemicals, suggesting that this is at least partially responsible for the resistance of this organism to these agents. Although Dictyostelium is also unusually resistant to high concentrations of H(2)O(2), little is known about the response of this organism to oxidative stress. To determine if transcriptional upregulation is a common mechanism for responding to DNA-damaging agents, we have studied the Dictyostelium catalase and Cu/Zn superoxide dismutase antioxidant enzymes. We show that there are two catalase genes and that each is differentially regulated both temporally and spatially during multicellular development. The catA gene is expressed throughout growth and development and its corresponding enzyme is maintained at a steady level. In contrast, the catB gene encodes a larger protein and is only expressed during the final stages of morphogenesis. Cell type fractionation showed that the CatB enzyme is exclusively localized to the prespore cells and the CatA enzyme is found exclusively in the prestalk cells. Each enzyme has a different subcellular localization. The unique developmental timing and cell type distribution suggest that the role for catB in cell differentiation is to protect the dormant spores from oxidative damage. We found that exposure to H(2)O(2) does not result in the induction of the catalase, superoxide dismutase, NER or BER mRNAs. A mutant with greatly reduced levels of catA mRNA and enzyme has greatly increased sensitivity to H(2)O(2) but normal sensitivity to UV. These results indicate that the natural resistance to oxidative stress is not due to an ability to rapidly raise the level of antioxidant or DNA repair enzymes and that the response to UV-light is independent from the response to reactive oxygen compounds. PMID- 11004504 TI - The genomic structure and promoter analysis of the human ABF-1 gene. AB - The human ABF-1 gene is expressed in activated B-cells and Epstein-Barr virus immortalized lymphoblastoid cell lines. ABF-1 represents the only member belonging to the basic helix-loop-helix (bHLH) family of transcription factors whose expression pattern is restricted to B-cells. ABF-1 forms heterodimeric complexes with E2A to modulate gene transcription. We report the cloning and characterization of the human ABF-1 gene and the promoter region. The gene spans more than 3 kb and contains two exons. Exon 1 contains 274 bp of a 5' untranslated sequence (UTR) while exon 2 contains 1097 bp of 3'-UTR. Promoter analysis of the 5'-flanking region revealed no apparent B-cell-restricted control elements within approximately 700 bp, but clearly demonstrated the presence of a functional minimal promoter residing immediately upstream of the transcription start site. Analysis of the region containing the minimal promoter activity identified no CCAAT or TATA sequence. Lastly, we have assigned the ABF-1 gene to human chromosome 8q21.1 using fluorescence in situ hybridization (FISH). The cloning of the human ABF-1 gene will facilitate further biochemical and genetic studies of its function in the regulation of B-cell differentiation. PMID- 11004505 TI - Novel responses of ZRF, a variant of human MTF-1, to in vivo treatment with heavy metals. AB - Heavy metal-dependent transcriptional activation of metallothionein (MT) genes is mediated by multiple enhancer sequences, metal responsive element (MRE), located in the upstream region of the genes. Previously, we have reported purification of a zinc-dependent MRE-binding protein, zinc regulatory factor (ZRF), from HeLa cells, and have pointed to the close relationship between ZRF and mouse MRE binding transcription factor-1 (MTF-1) according to the analysis of partial amino acid sequences. By means of cDNA cloning and the product analyses, we show that ZRF is a variant of human MTF-1 (hMTF-1), which carries a single amino acid exchange in the zinc finger domain. Accordingly, ZRF is renamed hMTF-1b. Expression of hMTF-1b in HeLa cells is constitutive at both mRNA and protein levels, and is unaffected by treatment with cadmium (Cd). On the other hand, when cells were fractionated into nuclear extract and cytosol, a large part of the hMTF-1b was recovered in the cytosol fraction. A significant increase in the amount of nuclear hMTF-1b occurs when cells are treated with various heavy metals, including Cd, Zn, Cu and Ag, which is associated with concomitant decrease in the amount recovered in the cytosol fraction. Since immunocytochemical analysis revealed that intracellular distribution of hMTF-1b is restricted to the nucleus irrespective of the heavy metal treatment, such an increment in the nuclear extracts apparently results from promotion of nuclear retention of hMTF-1b by the heavy metal treatment. Analysis by native gel electrophoresis shows that the mobility of hMTF-1b significantly changes in association with Cd treatment, raising the possibility that a conformational change of hMTF-1b occurs in response to treatment with heavy metals in vivo. PMID- 11004506 TI - Transcription of the catalytic 180-kDa subunit gene of mouse DNA polymerase alpha is controlled by E2F, an Ets-related transcription factor, and Sp1. AB - We have isolated a genomic DNA fragment spanning the 5'-end of the gene encoding the catalytic subunit of mouse DNA polymerase alpha. The nucleotide sequence of the upstream region was G/C-rich and lacked a TATA box. Transient expression assays in cycling NIH 3T3 cells demonstrated that the GC box of 20 bp (at nucleotides -112/-93 with respect to the transcription initiation site) and the palindromic sequence of 14 bp (at nucleotides -71/-58) were essential for basal promoter activity. Electrophoretic mobility shift assays showed that Sp1 binds to the GC box. We also purified a protein capable of binding to the palindrome and identified it as GA-binding protein (GABP), an Ets- and Notch-related transcription factor. Transient expression assays in synchronized NIH 3T3 cells revealed that three variant E2F sites near the transcription initiation site (at nucleotides -23/-16, -1/+7 and +17/+29) had no basal promoter activity by themselves, but were essential for growth-dependent stimulation of the gene expression. These data indicate that E2F, GABP and Sp1 regulate the gene expression of this principal replication enzyme. PMID- 11004508 TI - Telomeric profiles and telomerase activity in turkey satellite cell clones with different in vitro growth characteristics. AB - The satellite cell population in postnatal skeletal muscle is heterogeneous because individual satellite cells isolated from a single muscle have differing abilities to proliferate under the same in vitro conditions. Telomeres are structures found at the ends of all eukaryotic chromosomes that are characterized by repetitive DNA sequences, and they are important in determining cellular proliferation potential. The relationship between satellite cell proliferative heterogeneity and telomeric DNA was examined by digesting genomic DNA from large colony-forming and small-colony-forming turkey satellite cell clones with HinfI, separating the restriction fragments on an agarose gel, and hybridizing the gels with an oligonucleotide probe specific for telomeric DNA. Turkey satellite cells generated telomeric restriction fragments up to approximately 180 kB. The large colony-forming satellite cell clones had a larger proportion (P<0.05) of total telomeric restriction fragments below 33 kB than the small-colony-forming satellite cell clones. However, telomerase expression was detected in cultures from large-colony-forming and small-colony-forming turkey satellite cells suggesting that the differences in telomeric restriction fragments may not be related to the differences in in vitro proliferative behavior and that telomerase may contribute to the high in vitro growth capacity of turkey satellite cells. PMID- 11004507 TI - Structure and genetic polymorphism of the mouse KCC1 gene. AB - The KCC1 K-Cl cotransporter is a major regulator of erythroid and non-erythroid cell volume, and the KCC1 gene is a candidate modifier gene for sickle cell disease and other hemoglobinopathies. We have cloned and sequenced the mouse KCC1 (mKCC1) gene, defined its intron-exon junctions, and analyzed (AC)/(TG) intragenic polymorphisms. A highly polymorphic (AC) repeat of mKCC1 intron 1 was characterized in musculus strains, and used to prove lack of linkage between the mKCC1 gene and the rol (resistant to osmotic lysis) locus. The intron 1 (AC) repeat in CAST/Ei and SPRET/Ei was not only more divergent in length but also underwent additional sequence variation. A dimorphic (TG) repeat in intron 2 distinguished CAST/Ei from other strains, and an intron 17 B1 Alu-like SINE present in all musculus strains was found to be absent from intron 17 in SPRET/Ei. These and additional described strain-specific polymorphisms will be useful mapping and genetic tools in the study of mouse models of sickle cell disease. PMID- 11004509 TI - Functional cloning and mutational analysis of the human cDNA for phosphoacetylglucosamine mutase: identification of the amino acid residues essential for the catalysis. AB - In Saccharomyces cerevisiae, phosphoacetylglucosamine mutase is encoded by an essential gene called AGM1. The human AGM1 cDNA (HsAGM1) and the Candida albicans AGM1 gene (CaAGM1) were functionally cloned and characterized by using an S. cerevisiae strain in which the endogenous phosphoacetylglucosamine mutase was depleted. When expressed in Escherichia coli as fusion proteins with glutathione S-transferase, both HsAgm1 and CaAgm1 proteins displayed phosphoacetylglucosamine mutase activities, demonstrating that they indeed specify phosphoacetylglucosamine mutase. Sequence comparison of HsAgm1p with several hexose-phosphate mutases yielded three domains that are highly conserved among phosphoacetylglucosamine mutases and phosphoglucomutases of divergent organisms. Mutations of the conserved amino acids found in these domains, which were designated region I, II, and III, respectively, demonstrated that alanine substitutions for Ser(64) and His(65) in region I, and for Asp(276), Asp(278), and Arg(281) in region II of HsAgm1p severely diminished the enzyme activity and the ability to rescue the S. cerevisiae agm1Delta null mutant. Conservative mutations of His(65) and Asp(276) restored detectable activities, whereas those of Ser(64), Asp(278), and Arg(281) did not. These results indicate that Ser(64), Asp(278), and Arg(281) of HsAgm1p are residues essential for the catalysis. Because Ser(64) corresponds to the phosphorylating serine in the E. coli phosphoglucosamine mutase, it is likely that the activation of HsAgm1p also requires phosphorylation on Ser(64). Furthermore, alanine substitution for Arg(496) in region III significantly increased the K(m) value for N acetylglucosamine-6-phosphate, demonstrating that Arg(496) serves as a binding site for N-acetylglucosamine-6-phosphate. PMID- 11004510 TI - Identification of cAMP analogue inducible genes in RAW264 macrophages. AB - RNA was isolated from RAW264 cells treated with or without 8-Br-cAMP and the differential display and subtractive hybridization methods were performed. One hundred and twenty-five differentially displayed bands were identified. Upon Northern blot analysis, only three of these bands were confirmed as cAMP inducible mRNAs, named cI-1, cI-2, and cI-3 (for cAMP inducible genes 1-3). The cI-3 probe was identical to a previously known gene, gly96. Using the novel cI-1 and cI-2 partial cDNAs as probes, a mouse macrophage cDNA library was screened and the two full length genes were cloned, sequenced, and characterized as encoding large hydrophobic proteins. One hundred and fifteen partial cDNA clones from a subtractive hybridization library were also screened by Northern blot and 64 were found to be cAMP inducible. Of these, 45 represented 31 known unique genes in the GenBank nr database (cI-4-34), and 19 clones representing 15 unique sequences were not in the nr database (cI-35-49). One of the previously known genes was ABC1, the Tangier disease gene, which was identified from four independent partial cDNAs. ABC1 was upregulated in RAW cells by cAMP, concurrent with the cAMP induction of lipid efflux to apolipoprotein A1. PMID- 11004511 TI - Nucleotide sequence, genomic organization and cell-cycle-dependent expression of a Chlamydomonas 14-3-3 gene. AB - Members of the 14-3-3 protein family have been identified as regulatory elements in intracellular signalling pathways and cell cycle control. Previously we reported the nucleotide sequence of a 14-3-3 cDNA cloned from the unicellular green alga Chlamydomonas reinhardtii. In this communication, we describe the nucleotide sequence, the genomic organization and the cell-cycle-dependent expression of the corresponding gene. The coding sequence of this gene was found to be interrupted by four introns of 124, 116, 81, and 659 bp, respectively. Introns 2-4 were found in conserved positions as compared to the Arabidopsis 14-3 3 genes. A counterpart to intron 1 absent in the Arabidopsis 14-3-3 genes was found in the human 14-3-3 epsilon gene. PMID- 11004512 TI - Spatiotemporal regulation of hnRNP M and 2H9 gene expression during mouse embryonic development. AB - Using the HeLa cell model along with an in vitro splicing system, we have previously shown that hnRNP M and 2H9 are involved in the pre-mRNA splicing process and most interestingly also in heat shock-induced transient splicing arrest by transiently leaving the hnRNP complexes. Due to this unique regulatory function in a mechanism that turns splicing on and off, these two hnRNPs appear as important proteins for controlling gene expression. Here we investigated by in situ hybridization and immunohistochemical staining techniques the expression level of specific mRNA and protein during mouse embryonic development. HnRNP M and 2H9 are found to be expressed at all examined stages (6.5-18.5 days post coitum), in a differential manner, and at various levels depending on tissues, cell types and also embryonic stages; fairly high levels of both hnRNPs are always observed in the central nervous system. Furthermore, levels of colocalizing protein and transcript are not always present in the same proportion, thus suggesting a post-transcriptional regulation of hnRNP M and 2H9 gene expression. The complex spatiotemporal variations we observed might well anticipate a role for these two hnRNPs also in modulating splicing, thereby influencing gene expression and further many physiological processes. PMID- 11004513 TI - Human CKIalpha(L) and CKIalpha(S) are encoded by both 2.4- and 4. 2-kb transcripts, the longer containing multiple RNA-destablising elements. AB - Casein kinase I (CKI) are a family of conserved second messenger-independent serine/threonine protein kinases found in all eukaryotes. The avian and mammalian CKI alpha isoform has four splice variants differing in the presence or absence of 28 amino acids ('(L)' insertion) in the catalytic domain and/or 12 amino acids ('(S)' insertion) in the regulatory domain. Here we report the isolation of cDNAs encoding human CKIalpha(L) and CKIalpha(S). We find human CKIalpha(L) has a preference to phosphorylate phosvitin over casein, with a higher K(m) for casein than phosvitin, the reverse being the case for human CKIalpha(S). Both human CKIalpha(L), and CKIalpha(S) are derived from 4.2-kb mRNA transcripts and 2.4-kb transcripts, the latter probably generated by use of an alternate polyadenylation signal identified in the longer transcripts. The 4. 2-kb transcripts contain six RNA-destabilising AU-rich element (ARE) motifs in the 3'-untranslated region (UTR), while the 2.4-kb transcripts contain a single ARE motif. In vitro analysis of CKI alpha 3'-UTR RNA sequences suggests that in HeLa cells, the longer 3'-UTR transcripts are likely to degrade approximately 13 times faster than the shorter 3'-UTR transcripts. This is the first report of a kinase mRNA containing multiple RNA-destabilising AREs in the longer of two mRNA transcripts. PMID- 11004514 TI - Characterisation of the rat heparin-binding epidermal growth factor-like growth factor gene promoter. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) gene expression is strongly activated by a variety of extracellular stimuli, acting through the Raf/MEK/MAP kinase pathway. To study the elements that respond to this pathway, we have isolated and sequenced a fragment of the rat HB-EGF gene promoter. By transfection of a series of promoter/reporter constructs into cells, a minimal promoter element was demonstrated to lie between 448 bp upstream of the transcriptional start site and 103 bp into the first exon of the gene. However co transfection of the promoter constructs with a plasmid directing expression of RafCAAX, an activated c-Raf-1 protein, gave a fold-stimulation of activity no greater than that seen for the parental pGL3-Basic plasmid alone. In addition, agonist stimulation of cell lines stably transfected with a HB-EGF promoter/luciferase construct produced little or no increase in reporter enzyme activity. These results suggest that the c-Raf-1 responsive elements lie outside the tested region of the rat HB-EGF gene. However, it has been reported that a c Raf-1 responsive element is present within the equivalent region of the mouse gene. A comparison of the 5'-flanking regions of the mouse, rat and human HB-EGF genes indicated that the mouse sequence diverges abruptly from that of the other two species approximately 260 bp upstream of the transcriptional start site. PCR analysis of mouse genomic DNA suggests that this sequence divergence is due to DNA rearrangement during the cloning of the mouse gene. Additional studies are therefore required to identify Raf/MAP kinase responsive elements in the HB-EGF gene. PMID- 11004515 TI - Genomic cloning, mapping, structure and promoter analysis of HEADPIN, a serpin which is down-regulated in head and neck cancer cells. AB - Headpin is a novel serine proteinase inhibitor (serpin) that is down-regulated in squamous cell carcinoma of the oral cavity and in squamous cell carcinoma cell lines of the head and neck. Using a panel of 18q21.3 YAC clones, we mapped and cloned the HEADPIN gene. The gene spans 10 kb and is composed of eight exons and seven introns. The genomic structure is identical with some other ovalbumin serpins (ov-serpins) in terms of the numbers, position and phasing of the intron/exon boundaries. HEADPIN was mapped within the serpin cluster in 18q21.3 between MASPIN and SCCA2 as follows: cen-MASPIN-HEADPIN-SCCA2-SCCA1-tel. The transcription start site was determined and the promoter activity of the 5' flanking region was analyzed. Luciferase promoter assays in HaCaT cells showed that the -432 to -144 nucleotide region has functional promoter activity. The activity of the promoter/enhancer was not observed in head and neck cancer cell lines TU167 and UMSCC1 which lack headpin expression. These data suggest that the differential expression of headpin in normal and carcinoma-derived cells is regulated at the transcriptional level. Understanding the genomic organization and transcriptional regulation of the ov-serpins clustered within 18q21. 3 provides a critical framework for assessing their potential role in cancer. PMID- 11004516 TI - Cloning and gene expression of a novel human ribonucleoprotein. AB - This paper reports on the cloning and characterization of a novel human ribonucleoprotein, RBM8, containing a single RNA binding domain comprising the two RNP-CS and RNP-2 consensus motifs. The protein has 55% identity to a segment of a C. elegans ribonucleoprotein of unknown function. The RBM8 gene shows ubiquitous tissue expression, predominantly as a 0.9 kb transcript. An interesting feature of the RBM8 transcript is an homology of 42% in the 3' untranslated region, in the antisense orientation, to the human gonadotropin releasing hormone receptor polypeptide. RBM8 maps to human chromosome 14 in the 14q21-q23 region. PMID- 11004517 TI - Sjalpha elements, short interspersed element-like retroposons bearing a hammerhead ribozyme motif from the genome of the oriental blood fluke Schistosoma japonicum. AB - Smalpha is a short interspersed element (SINE)-like retroposon that occurs in high copy number of the genome of the human blood fluke Schistosoma mansoni. The sequence of the consensus Smalpha element includes the hallmark features of SINE like elements including a promoter region for RNA polymerase III, an AT-rich stretch at its 3'-terminus, a short length of 500 bp or less, and short direct repeat sequences flanking the insertion site. Interestingly, the sequence of Smalpha also encodes an active ribozyme bearing a hammerhead domain. Contrary to the recent findings of Ferbeyre et al. (Mol. Cell. Biol. 18 (1998) 3880-8) that indicated that Smalpha-like elements were absent from the genome of the Oriental blood fluke Schistosoma japonicum, we report here that the genome of S. japonicum does contain a family of Smalpha-like retroposons, elements that we have named the Sjalpha family. Like Smalpha, Sjalpha elements are SINE-like in structure and sequence, are present at high copy number interspersed throughout the S. japonicum genome, and contain an ostensibly functional, hammerhead ribozyme motif. The presence of these elements in all species of Schistosoma so far examined suggests that the hammerhead domain was acquired by vertical transmission from a common schistosome ancestor. PMID- 11004518 TI - Characterization of mouse cathepsin R, a new member of a family of placentally expressed cysteine proteases. AB - A new mouse cysteine protease, termed cathepsin R, has been identified. The complete nucleotide sequence of this gene was derived from a set of cDNAs generated from 15.5-day mouse placenta. Sequence analysis revealed an open reading frame encoding a 334 amino acid long polypeptide closely related to placentally expressed cathepsins P, Q, and M. RT-PCR analysis indicated that cathepsin R is only expressed in placenta and thus is a new member of the emerging family of cathepsins whose expression is regulated during mouse embryonic development. Modeling and structural analysis suggests that cathepsin R will have a restricted substrate specificity when compared to that of cathepsin L. PMID- 11004519 TI - Mouse connexin37: gene structure and promoter analysis. AB - Connexin37 (Cx37) is a subunit gap junction protein which exhibits limited expression in only a few cell types, predominantly in endothelial cells and in the lung. To begin to analyze Cx37 expression, we isolated a 1.6 kb mouse Cx37 cDNA from a mouse lung cDNA library and isolated corresponding mouse genomic clones from a bacterial artificial chromosome library. Sequencing and comparison of these clones showed that the Cx37 gene contained a short first exon, an 1.0 kb single intron and a second exon containing the complete coding region and 3' untranslated region (UTR). The 5'-UTR of the mouse cDNA showed 70% identity to that of human Cx37. Primer extension experiments performed using mouse lung RNA gave two bands of sizes consistent with the transcription start site predicted from the cDNA. Sequence analysis showed that the regions flanking exon I contained a consensus 'TATA box' 43 bp 5' from the transcription start site preceded by several putative transcription factor binding sites and a 282 bp truncated L1Md interspersed element. Luciferase reporter gene transfections suggested that an area of 268 bp 5' from the first exon acted as a basal promoter for Cx37 and that there was a strong negative regulatory element in the intron. PMID- 11004520 TI - Identification of a composite enhancer of the human tyrosinase-related protein 2/DOPAchrome tautomerase gene. AB - The human tyrosinase-related protein 2 (TRP-2) gene promoter contains a cis regulatory element (positions -447 to -416), termed DOPAchrome tautomerase distal enhancer 1 (DDE1). DDE1 functions as an enhancer in cultured melanoma cells and its core element includes a potential binding site for transcription factors containing a high-mobility-group domain. This core element is bound in vitro by multiple nuclear proteins, which are preferentially expressed in melanoma cells. DDE1 represents a composite enhancer that may be involved in melanocyte-specific transcription of the human TRP-2 gene. PMID- 11004521 TI - A novel proteinase inhibitor gene transiently induced by tobacco mosaic virus infection. AB - A gene (NgPI) encoding a novel proteinase inhibitor (PI) has been isolated from tobacco leaves. Protein encoded by the gene consists of 241 amino acid residues having a predicted molecular mass of 26.7 kDa and a calculated pI of 8.7. A predicted N-terminal signal sequence followed by a vacuolar targeting signal and a peptide conserved in the Kunitz type PIs were identified. The deduced NgPI protein has sequence homology with aspartic and cysteine protease inhibitors. The gene is present as double copies in the Nicotiana glutinosa genome. Expression of the NgPI gene is rapidly and transiently induced by tobacco mosaic virus infection at a time earlier than apparent lesions of hypersensitive responses appear on the leaves. PMID- 11004522 TI - Cloning and characterization of a novel human ninein protein that interacts with the glycogen synthase kinase 3beta. AB - Using human glycogen synthase kinase 3beta (GSK-3beta) as bait in the yeast two hybrid system, we identified a novel human centrosome associated protein, hNinein. When the full length cDNA of hNinein was sequenced, it showed that an open reading frame encoded a protein consisting of 2047 amino acids with a predicted molecular mass of 239 kDa. The features of this protein include a potential GTP binding site, a large coiled-coil domain together with four leucine zipper domains and a GSK-3beta binding site. Fluorescence microscopy experiment showed that hNinein is localized in the pericentriolar matrix of the centrosome. In addition, hNinein also showed to react with centrosomal autoantibody sera. Our findings suggest that hNinein may be involved in the formation of centrosome matrix and interacts with the GSK-3beta, implying that it may also be regulated by GSK-3beta phosphorylation signaling. PMID- 11004523 TI - Cloning of Z39Ig, a novel gene with immunoglobulin-like domains located on human chromosome X. AB - The cDNA sequence and expression profile of a novel human gene, encoding a new member of the immunoglobulin superfamily, is reported. The gene is localized in the pericentromeric region of human X chromosome between the markers DXS1213 and DXS1194. Abundant expression of transcripts was detected in several human fetal tissues, whereas among adult tissues lung and placenta express highest levels of Z39Ig mRNA. PMID- 11004524 TI - Molecular cloning of the murine cMOAT ATPase. AB - cMOAT encodes an ATPase within the family of cMOAT/MRP ATPases that functions as an ATP dependent, multispecific anion transporter within the canalicular surface of hepatocytes that has pharmacologic significance. We describe here the cloning of a murine cMOAT cDNA isolated from mouse liver. The open reading frame of this cDNA incorporates 4627 nucleotides encoding 1309 amino acids with 77.5% and 86.7% identity with the human and rat encoded amino acids, respectively. Northern blotting showed that the expression of cMOAT mRNA occurs primarily in mouse liver in the form of two variants with approximately 5.6 and 7.8 kb of sequence each. cMOAT mRNA was also detected in mouse kidney at a low level but was not detected in other mouse organs or tumors except the Hep 1-6 murine hepatoma where expression was also in the form of the same two mRNA variants. PMID- 11004525 TI - Cis element 'decoy' against the upstream promoter of the human estrogen receptor gene. AB - It is well known that breast carcinomas without estrogen receptor (ER) have a poor prognosis and do not respond to endocrine therapy. In analyzing the question of the lack of ER gene expression, we have considered the possibility that specific negative transcription factors are present in ER-negative breast cancers. Inside the P3 upstream promoter of human ER gene we identified a transcriptional regulatory sequence able to bind protein factors expressed in ER negative MDA-MB-231 breast cancer cells. This sequence, lying between nucleotides -3258 to -3157, seems to be critical for inhibition of ER gene transcription. In fact, the selected sequence in the form of double-stranded DNA has been introduced into ER-negative breast cancer cells as 'decoy' cis elements showing the ability to remove the putative negative transcription factor(s) and to induce the reactivation of ER gene transcription. In addition, in transient transfection assays the selected sequence decreased the SV-40 promoted luciferase activity. Gel shift assays identified multiple DNA-protein interactions which specifically form in this region, and data from Southwestern experiments strongly suggested the presence of a specific protein expressed in MDA-MB-231 ER-negative, but not in MCF7 ER-positive cells. PMID- 11004526 TI - Posttranslational processing and differential glycosylation of Tractin, an Ig superfamily member involved in regulation of axonal outgrowth. AB - Tractin is a novel member of the Ig-superfamily which has a highly unusual structure. It contains six Ig domains, four FNIII-like domains, an acidic domain, 12 repeats of a novel proline- and glycine-rich motif with sequence similarity to collagen, a transmembrane domain, and an intracellular tail with an ankyrin and a PDZ domain binding motif. By generating domain-specific antibodies, we show that Tractin is proteolytically processed at two cleavage sites, one located in the third FNIII domain, and a second located just proximal to the transmembrane domain resulting in the formation of four fragments. The most NH(2)-terminal fragment which is glycosylated with the Lan3-2, Lan4-2, and Laz2-369 glycoepitopes is secreted, and we present evidence which supports a model in which the remaining fragments combine to form a secreted homodimer as well as a transmembrane heterodimer. The extracellular domain of the dimers is mostly made up of the collagen-like PG/YG-repeat domain but also contains 11/2 FNIII domain and the acidic domain. The collagen-like PG/YG-repeat domain could be selectively digested by collagenase and we show by yeast two-hybrid analysis that the intracellular domain of Tractin can interact with ankyrin. Thus, the transmembrane heterodimer of Tractin constitutes a novel protein domain configuration where sequence that has properties similar to that of extracellular matrix molecules is directly linked to the cytoskeleton through interactions with ankyrin. PMID- 11004527 TI - Thioxo amino acid pyrrolidides and thiazolidides: new inhibitors of proline specific peptidases. AB - Aminopeptidase P (APP), dipeptidyl peptidase II (DP II), dipeptidyl peptidase IV (DP IV) and prolyl oligopeptidase (POP) are proline specific peptidases. Hence, they are able to cleave peptide bonds containing the imino acid proline. Amino acid pyrrolidides (Pyrr) and thiazolidides (Thia) are well-known product analogue inhibitors of DP IV and POP. For the first time we describe the influence of a thioxo amide bond, incorporated into these compounds, on the inhibition of the proline specific peptidases. Taking into account the substrate specificity of these peptidases, we have synthesized Xaa-psi[CS-N]-Pyrr and Xaa-psi[CS-N]-Thia of the amino acids Ala, Phe, Val and Ile. The inhibition constants were determined for the above mentioned proline specific peptidases isolated from different sources. As a result, the serine proteases DP II, DP IV and POP were inhibited competitively, whereas metal-dependent APP displayed a linear mixed type inhibition with inhibition constants up to 10(-4) M. Thioxylation of Xaa Pyrr and Xaa-Thia led to a slight decrease of inhibition of DP IV and POP compared to Xaa-Pyrr and Xaa-Thia, though the inhibition constants were still in the range up to 10(-7) M. As Xaa-Thia exist as two isomers, we investigated isomer specific inhibition with regard to DP IV. Thus, our studies have revealed that DP IV was only inhibited by the Z isomer of the Xaa-psi[CS-N]-Thia. For the first time, Xaa-Pyrr and Xaa-Thia were characterized as inhibitors of DP II with inhibition constants in the micromolar range. In contrast to DP IV inhibition, the Xaa-psi[CS-N]-Pyrr and Xaa-psi[CS-N]-Thia have proven to be more potent inhibitors of DP II than the corresponding Xaa-Pyrr and Xaa-Thia. Thus, these Xaa psi[CS-N]-Thia are new potent inhibitors especially suitable for DP II with K(i) values ranging in the upper nanomolar concentration. PMID- 11004528 TI - Distinct physical and structural properties of the ovine uterine serpin. AB - Experiments were performed to examine the relationship between the structure and function of ovine uterine serpin (OvUS). Limited proteolytic digestion of OvUS caused cleavage of the 55-57 kDa OvUS to a 42 kDa product nearly identical in molecular weight to a naturally-occurring breakdown product of OvUS. N-terminal amino acid sequencing and MALDI-MS revealed that, unlike other serpins, OvUS was preferentially cleaved at about 70 amino acids upstream of the putative reactive center loop. Analysis of the partially-digested protein by gel filtration chromatography suggested that the C-terminal fragment of the protein was still associated under nondenaturing conditions. Partial digestion of OvUS had no effect on the protein's secondary structure, thermal stability, ability to bind lymphocytes or pepsin, or inhibitory activity towards pepsin or mitogen-induced lymphocyte proliferation. In contrast, mild denaturation of OvUS with 0.5 M guanidine HCl increased thermal stability. Unlike for other serpins, the increase in thermal stability was lost upon removal of the denaturant. Incubation of OvUS with 100 fold molar excess of a peptide corresponding to the putative P(14)-P(2) region of the RCL for 24 h at 37 degrees C to induce binary complex formation had no effect on its secondary structure and did not alter the biological activity of the protein. Synthetic peptides corresponding to the putative P(14)-P(2) region and the P(7)-P(15') region of the RCL were not inhibitory to pepsin activity or lymphocyte proliferation. Taken together, these results indicate that the conformation of OvUS is distinct from the prototypical serpin because conditions that lead to the large-scale conformational change in other serpins such as antithrombin III and alpha(1)-antitrypsin do not cause similar changes in OvUS. Moreover, the putative RCL does not seem to contain the activity required to inhibit lymphocyte proliferation or pepsin activity. PMID- 11004529 TI - Evidence for alternative binding modes in the interaction of benzylamine analogues with bovine liver monoamine oxidase B. AB - The interaction of purified bovine liver MAO B with the benzylamine analogues N,N dimethylbenzylamine and alpha-methylbenzylamine has been investigated. Both classes of analogues are competitive inhibitors of benzylamine oxidase activity. The K(i) values were determined for nine different para-substituted N, N dimethylbenzylamine analogues. Analysis of the binding affinities demonstrate the deprotonated forms of the tertiary amines are preferentially bound to MAO B and the affinity decreases with increasing van der Waals volume of the para substituent. The correlation for this relation is:Log K(i)=-0.97+/ (0.28)sigma+(0. 75+/-0.11)(0.1xV(w))-4.24+/-(0.16)alpha-Methyl benzylamine analogues are also found to be competitive inhibitors of MAO B-catalyzed benzylamine oxidation. Similar K(i) values were determined using either the S or R stereoisomers. Analysis of the binding affinities of five para-substituted alpha-methylbenzylamine analogues to MAO B shows the deprotonated form also to be preferentially bound and the affinity is marginally increased with increasing van der Waals volume of the para-substituent:Log K(i)=-0.71sigma-(0.32)(0. 1xV(w)) 3.50Comparison of these data with that previously published for para-substituted benzylamine binding to MAO B (Walker and Edmondson, Biochemistry 33 (1994) 7088 7098) demonstrates that these benzylamine analogues exhibit differing modes of binding to the active site of MAO B. The presence of an electronic substituent effect in the binding of these two classes of analogues compared with the lack of an observable electronic effect in the binding of benzylamine to MAO B is consistent with the proposal that orientation of the benzyl ring of the bound substrate is responsible for the absence of an electronic substituent effect on the rate of the reductive half reaction (Miller and Edmondson, Biochemistry 38 (1999) 13670-13683). PMID- 11004530 TI - Channeling efficiency in the bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase domain: the effects of site-directed mutagenesis of NADP binding residues. AB - The three-dimensional structure of the dehydrogenase-cyclohydrolase bifunctional domain of the human trifunctional enzyme indicates that Arg-173 and Ser-197 are within 3 A of the 2'-phosphate of bound NADP. Site-directed mutagenesis confirms that Arg-173 is essential for efficient binding and cannot be substituted by lysine. R173A and R173K have detectable dehydrogenase activity, but the K(m) values for NADP are increased by at least 500-fold. The S197A mutant has a K(m) for NADP that is only 20-fold higher than wild-type, indicating that it plays a supporting role. Forward and reverse cyclohydrolase activities of all the mutants were unchanged, except that the reverse cyclohydrolase activity of mutants that bind NADP poorly, or lack Ser-197, cannot be stimulated by 2',5'-ADP. The 50% channeling efficiency in the forward direction is not improved by the addition of exogenous NADPH and cannot be explained by premature dissociation of the dinucleotide from the ternary complex. As well, channeling is unaffected in mutants that exhibit a wide range of dinucleotide binding. Given that dinucleotide binding is unrelated to substrate channeling efficiency in the D/C domain, we propose that the difference in forward and reverse channeling efficiencies can be explained solely by the movement of the methenylH(4)folate between two overlapping subsites to which it has different binding affinities. PMID- 11004531 TI - Hydrolysis by plasma kallikrein of fluorogenic peptides derived from prorenin processing site. AB - Human plasma kallikrein (HPK) activates plasma prorenin to renin, and the physiological significance of this activation is still unknown. In this paper we investigated the efficiency and the cleavage pattern of the hydrolysis by HPK of the internally quenched fluorescent peptides (qf-peptides) derived from the amino acid sequence of human prorenin cleavage site. The peptide Abz-F-S-Q-P-M-K-R-L-T L-G-N-T-T-Q-EDDnp (Abz=ortho-aminobenzoic acid, and EDDnp=N-[2,4-dinitrophenyl] ethylene diamine), that corresponds to the amino acid sequence P(7) to P(7)' of human prorenin cleavage site, is hydrolyzed at the correct processing site (R-L bond) with k(cat)/K(m)=85 mM(-1) s(-1). Alanine was scanned in all positions from P(5) to P(5)' in order to investigate the substrate specificity requirements of HPK. The qf-peptides derived from the equivalent segment of rat prorenin, that has Lys-Lys as basic amino acid pair, and the peptide Abz-NVTSPVQ-EDDnp that contains the proposed cleavage site of rat prorenin have very low susceptibility to hydrolysis by rat plasma kallikrein. These data are according to the previously reported absence of rat plasma prorenin activation by rat plasma kallikrein (RPK), and with the view that prorenin activation in rat requires alternative enzymes and/or mechanism. All the obtained peptides described in this paper were also assayed with bovine trypsin that was taken as a reference protease because it is commonly used to activate prorenin. PMID- 11004532 TI - Glutamine synthetase, hemoglobin alpha-chain, and macrophage migration inhibitory factor binding to amyloid beta-protein: their identification in rat brain by a novel affinity chromatography and in Alzheimer's disease brain by immunoprecipitation. AB - Proteins binding to amyloid beta-protein (Abeta) may modulate the accumulation of Abeta in Alzheimer's disease (AD) brain. We developed a monomeric Abeta column for isolation of the proteins binding to Abeta from rat brain. By amino acid sequence analysis and immunoreactivity with specific antibodies, we identified three new Abeta-binding proteins, glutamine synthetase, hemoglobin alpha-chain, and macrophage migration inhibitory factor as well as serum albumin, beta tubulin, and glyceraldehyde-3-phosphate dehydrogenase already identified as proteins bound to amyloid beta-protein precursor. In addition, the retained fraction contained both apolipoprotein E and alpha(1)-antichymotrypsin already known as Abeta binding proteins. Furthermore, we detected the complexes of these new binding proteins with Abeta in a soluble fraction of the cerebral cortex of AD brain by immunoprecipitation. Our results suggest that these binding proteins also associate with Abeta, leading to the clearance or the accumulation of Abeta and the neuronal cell damage in human brain. PMID- 11004533 TI - Role of salt bridge(s) in the binding and photoconversion of bilirubin bound to high affinity site on human serum albumin. AB - The role of salt bridge(s) (between epsilon-NH(2) groups of lysine residues of human serum albumin (HSA) and carboxyl groups of bilirubin) in the binding and photoconversion of bilirubin bound to high affinity site on HSA was investigated by covalent modification of approximately 20% internal (buried) lysine residues of HSA with acetic anhydride, succinic anhydride and O-methylisourea and white light irradiation of their complexes with bilirubin. The different HSA derivatives, namely, acetylated HSA (aHSA), succinylated HSA (sHSA) and guanidinated HSA (gHSA), thus obtained, were found to be homogeneous with respect to charge and size and characterized in detail in terms of mean residue ellipticity, Stokes radius, tryptophan fluorescence, bilirubin binding and the photochemistry of their complexes with bilirubin. All the three derivatives retained helical contents and molecular size (Stokes radius) similar to HSA except for sHSA which showed a slight increase in the Stokes radius from 3.56 to 3.64 nm. Further, fluorescence properties of aHSA and sHSA were also found to be different from HSA and gHSA. Based on difference spectral change, fluorescence quenching and fluorescence enhancement results of bilirubin bound to HSA and its derivatives, nearly 46 and 48% reduction in bilirubin binding was observed in the case of aHSA and sHSA, respectively. Both aHSA and sHSA showed a decrease of 8- and 10-fold, respectively, in association constant compared to native HSA. Although the bisignate circular dichroism (CD) spectra of an equimolar (1:1) bilirubin-HSA complex was retained by all three HSA derivatives, the intensity of both positive and negative CD Cotton effects decreased significantly in both aHSA and sHSA. gHSA which retained net charge identical to native HSA, showed little decrease in bilirubin binding and the intensity of bisignate CD Cotton effects. The photochemical reaction of bilirubin bound to aHSA and sHSA produced opposite results to those observed with HSA and gHSA. A brief (2 min) irradiation of an equimolar complex of bilirubin with both aHSA and sHSA accompanied a rapid shift (14-15 nm) in the absorption spectrum of the bound pigment towards the blue region and almost complete elimination of negative CD Cotton effects while only moderately affecting the magnitude of positive CD Cotton effects. On the other hand, similar treatment of the complexes of bilirubin with HSA and gHSA did not show any change in the absorption spectrum, only a slight decrease in the intensity of both positive and negative CD Cotton effects was observed. The fluorescence intensity of bilirubin bound to HSA and gHSA was increased upon irradiation with white light and after 30 min it was nearly twice the value observed at 0 min irradiation. Interestingly, no change in the fluorescence intensity of bilirubin bound either to aHSA or sHSA was observed upon irradiation, even on increasing the duration of irradiation to 1 h. Taken together, the results on fluorescence quenching, fluorescence enhancement, CD spectral changes and visible absorption spectroscopy suggest that salt bridge(s) of the type (-COO(-).(+)H(3)N-) in which the epsilon-NH(2) group(s) contributed by lysine residues, are not only involved in the enantioselective binding of bilirubin but also in the stereospecific photoisomerization of bilirubin bound to a high affinity site on HSA. PMID- 11004534 TI - Engineering of substrate mimetics as novel-type substrates for glutamic acid specific endopeptidases: design, synthesis, and application. AB - This account reports on the development and function of novel substrate mimetics as artificial substrates for Glu-specific endopeptidases. Firstly, in an empirical way, various aliphatic and aromatic analogs of the already established carboxymethyl thioester-substrate mimetics were designed from simple structure function relationship studies. The specificity of the newly developed substrates for Staphylococcus aureus V8 protease-catalyzed reactions have been examined by steady-state hydrolysis kinetic studies. Additionally, these studies were expanded to the use of the equally Glu-specific endopeptidase from Bacillus licheniformis (BL-GSE) which can easily be purified from alcalase in high yields. Finally, the novel substrate mimetics were used as acyl donor components in BL GSE- and V8 protease-catalyzed model acyl transfer reactions. The results clarify the newly developed substrate mimetics as efficient acyl donors as well as BL-GSE as an attractive alternative to V8 protease for enzymatic peptide synthesis. PMID- 11004535 TI - Expression and characterisation of a highly repetitive peptide derived from a wheat seed storage protein. AB - The high molecular weight (HMW) subunit group of wheat seed storage proteins impart elasticity to wheat doughs and glutens. They consist of three domains: non repetitive N- and C-terminal domains, which contain cysteine residues for covalent cross-linking, and a central domain consisting of repeated sequences. The circular dichroism and infrared (IR) spectra of an intact HMW subunit were compared with those of a peptide corresponding to the central repetitive domain expressed in Escherichia coli. This allowed the structure of the central domain to be studied in the absence of the N- and C-terminal domains and the contributions of these domains to the structure of the whole protein to be determined. In solution the peptide showed the presence of beta-turns and polyproline II-like structure. Variable temperature studies indicated an equilibrium between these two structures, the polyproline II conformation predominating at low temperatures and the beta-turn conformation at higher temperatures. IR in the hydrated solid state also indicated the presence of beta turns and intermolecular beta-sheet structures. In contrast, spectroscopy of the whole subunit showed the presence of alpha-helix in the N- and C-terminal domains. The content of beta-sheet was also higher in the whole subunit, indicating that the N- and C-terminal domains may promote the formation of intermolecular beta-sheet structures between the repetitive sequences, perhaps by aligning the molecules to promote interaction. PMID- 11004536 TI - PrP(c) mRNA, but not PrP(Sc) is found in the salivary glands of scrapie-infected sheep. AB - Transmission studies in transmissible spongiform encephalopathies (TSEs) have become increasingly important due to the possible transmission of bovine spongiform encephalopathy to humans resulting in new variant Creutzfeldt-Jacob disease. The horizontal transmission of scrapie, a TSE of sheep, is poorly understood. Possible sources of horizontal transmission are the submandibular and parotid salivary glands. TSEs like natural sheep scrapie are characterized by the conversion of a normal protease sensitive prion protein, PrP(c), to an abnormal protease resistant prion protein, PrP(Sc). Since the presence of PrP(Sc) is an indicator of disease, the salivary glands of scrapie-infected sheep were examined for the presence of PrP(Sc). Although PrP(c) mRNA was detected in the salivary glands, PrP(Sc) was not found in the salivary glands of scrapie-infected sheep. These data suggest that the salivary glands are unlikely sources of horizontal transmission of natural sheep scrapie. PMID- 11004537 TI - A new alternative method to quantify residual structure in 'unfolded' proteins. AB - Pig (pCSD1) and human (hCSD1) calpastatin domain 1 proteins were studied to characterize common features of the denatured state of proteins. These proteins were chosen for the present investigation, because pCSD1 was suggested previously to be unstructured in water even at 25 degrees C (1) [T. Konno et al., Biochim. Biophys. Acta 1342 (1997) 73-82]. hCSD1 could be expected to exhibit similar features on the basis of preliminary spectroscopic studies. In the present study, the experimental grounds for the estimate of residual structure in the unfolded state were differential scanning calorimetry heat capacity and circular dichroism (CD) measurements over the temperature range 10-80 degrees C. At selected temperatures, we studied also the effect of guanidinium hydrochloride (GdnHCl) which is known to promote further unfolding of the polypeptide chain. All other measurements were performed at pH 6 in pure water. The present results support the conclusion that the comparison of the experimentally obtained heat capacity data with theoretical heat capacity values calculated on the basis of a newly established increment system gives insight into the degree of hydration of the unfolded polypeptide chain. The percentage by which the experimental heat capacity of the unfolded polypeptide chain differs from the calculated heat capacity permits a quantitative estimate of the residual structure. This estimate is in good agreement with that based on CD absorption. The heat capacity approach has the advantage of comparing fully hydrated and partially hydrated residues in the same aqueous environment, whereas for example spectroscopic measurements, such as CD, are generally referred to the fully unfolded chain in concentrated urea or GdnHCl solutions. As the unfolded chains of pCSD1 and hCSD1 exhibit a smaller heat capacity than that calculated on the new peptide-based increment system [M. Hackel et al., J. Mol. Biol. 291 (1999) 197-213], we conclude that the residues in the unfolded polypeptide chain are less hydrated than the same residues in oligopeptides. This suboptimal hydration is the result of residual structure in the chain as observed in both CD and heat capacity measurements. PMID- 11004538 TI - Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyltransferase: pre steady-state kinetic studies. AB - Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyltransferase (DMAPP tRNA transferase) catalyzes the first step in the biosynthesis of the hypermodified A37 residue in tRNAs that read codons beginning with uridine. The mechanism of the enzyme-catalyzed reaction was studied by isotope trapping, pre steady-state rapid quench, and single turnover experiments. Isotope trapping indicated that the enzyme.tRNA complex is catalytically competent, whereas the enzyme.DMAPP complex is not. The results are consistent with an ordered sequential mechanism for substrate binding where tRNA binds first. The association and dissociation rate constants for the enzyme.tRNA binary complex are 1. 15+/-0.33x10(7) M(-1) s(-1) and 0.06+/-0.01 s(-1), respectively. Addition of DMAPP gives an enzyme.tRNA.DMAPP ternary complex in rapid equilibrium with the binary complex and DMAPP. Rapid quench studies yielded a linear profile (k(cat)=0.36+/-0.01 s(-1)) with no evidence for buildup of enzyme-bound product. Product release from DMAPP-tRNA transferase is therefore not rate-limiting. The Michaelis constant for tRNA and the equilibrium dissociation constant for DMAPP calculated from the individual rate constants determined here are consistent with values obtained from a steady-state kinetic analysis. PMID- 11004539 TI - Arrangement of functional units within the Rapana thomasiana hemocyanin subunit RtH2. AB - For the determination of the number and linear sequential arrangement of functional units (FUs) within the polypeptide chain of the Rapana hemocyanin subunit RtH2, a panel of mono-, di-, tri- and penta-FU fragments was generated by limited proteolysis of the purified subunit with four different enzymes. The individual cleavage products were isolated, characterized by SDS-PAGE and N terminally sequenced. Most of the information about the FU sequential arrangement within RtH2 was obtained after limited proteolysis of the subunit with plasmin. Overall correlation of the data revealed the sequential order of the eight FUs within the polypeptide chain of RtH2, termed RtH2-a to RtH2-h. The sites, most sensitive to proteolytic cleavage with plasmin, are located at the C-terminus, between the FUs ef, fg and gh. A second main cleavage site was observed between the FUs cd. Endoproteinase GluC hydrolyzes these sites, too, but produces exclusively a mixture of mono-, di- and tri-FU fragments. The most stable fragments, the trimer abc and the dimer gh, are found in all cleavage mixtures of RtH2 studied. RtH2-h is compared with the corresponding h-FUs of the gastropodan hemocyanins of Pila leopoldvillensis, Helix pomatia, Megathura crenulata and Haliotis tuberculata, and a remarkable similarity is observed between them: an increased M(r) of approximately 65000 instead of approximately 50000, estimated for an average FU, suggesting that the sequence of RtH2-h is elongated by about 95 amino acid residues at the C-terminal part of the molecule, as found for beta(c)-HpH, HtH1 and HtH2. PMID- 11004540 TI - Kinetic and hydrodynamic studies of the NodL O-acetyl transferase of Rhizobium leguminosarum: a random-order ternary complex mechanism for acetyl transfer by a roughly spherical trimeric protein. AB - The nodL gene product of Rhizobium leguminosarum is required for O-acetylation of diffusible lipo-oligosaccharide signalling factors which are involved in the host specific nodulation of legume roots. Kinetic studies of the forward reaction, using the substrate analogues chitosan pentaose and chitosan tetraose and the acyl donors acetyl-CoA and propionyl-CoA, and the dead-end inhibitor EtCoA are consistent with a steady-state random-order ternary complex mechanism in which the off rate of the O-acetyl chitosan oligomer appears to be partially rate determining. Moreover, the linearity of primary double-reciprocal plots favours the view that the interconversion of the ternary complex of NodL and its substrates with that of enzyme and bound products is not significantly faster than k(cat). Dissociation constants for coenzyme A and acetyl-CoA were determined by titration microcalorimetry to be 16.5 and 7.2 microM respectively, the latter in agreement with the kinetically derived value of 7.0 microM. The physical state of purified NodL, as determined by equilibrium centrifugation, velocity sedimentation and quasi-elastic light scattering, is that of a roughly spherical, trimeric protein with little tendency to self-associate. PMID- 11004541 TI - Conformational correlation and coupled motion between residue A21 and B25 side chain observed in crystal structures of insulin mutants at position A21. AB - The C-terminal residue of the insulin A chain is invariant and kept as asparagine in all known insulin molecules from hagfish through birds to mammals. To get information on the role of this conserved residue, which is still unclear, the three-dimensional structures of four human insulin mutants, A21 Asn-->Gly, A21 Asn-->Asp, A21 Asn-->Ala, and A21 Asn-->Gln DesB30, were determined by X-ray crystallography. The four mutants crystallize separately into two kinds (rhombohedral and cubic) of crystals. In the refined structures, conformational correlation and coupled motion between the A chain C-terminal residue A21 and the B25 side chain was observed, in contrast to the nearly unchanged general structures as compared with the native insulin structures in their respective crystals. A detailed analysis suggests that residue A21 can affect insulin receptor binding by interaction with the B25 side chain and the B chain C terminal segment to assist the B25 side chain rearranging into the 'active' conformation. PMID- 11004542 TI - Calcium-induced flexibility changes in the troponin C-troponin I complex. AB - The contraction of vertebrate striated muscle is modulated by Ca(2+) binding to the regulatory protein troponin C (TnC). Ca(2+) binding causes conformational changes in TnC which alter its interaction with the inhibitory protein troponin I (TnI), initiating the regulatory process. We have used the frequency domain method of fluorescence resonance energy transfer (FRET) to measure distances and distance distributions between specific sites in the TnC-TnI complex in the presence and absence of Ca(2+) or Mg(2+). Using sequences based on rabbit skeletal muscle proteins, we prepared functional, binary complexes of wild-type TnC and a TnI mutant which contains no Cys residues and a single Trp residue at position 106 within the TnI inhibitory region. We used TnI Trp-106 as the FRET donor, and we introduced energy acceptor groups into TnC by labeling at Met-25 with dansyl aziridine or at Cys-98 with N-(iodoacetyl)-N'-(1-sulfo-5 naphthyl)ethylenediamine. Our distance distribution measurements indicate that the TnC-TnI complex is relatively rigid in the absence of Ca(2+), but becomes much more flexible when Ca(2+) binds to regulatory sites in TnC. This increased flexibility may be propagated to the whole thin filament, helping to release the inhibition of actomyosin ATPase activity and allowing the muscle to contract. This is the first report of distance distributions between TnC and TnI in their binary complex. PMID- 11004543 TI - Effect of pH on the structure and aggregation of human glycodelin A. A comparison with beta-lactoglobulin A. AB - The effect of pH on the structure of glycodelin A (GdA) and of beta-lactoglobulin A (beta-LgA) has been investigated by means of circular dichroism, steady state fluorescence, synchrotron radiation small angle X-ray scattering (SR-SAXS) and gel permeation chromatography. The comparison between GdA and beta-LgA shows that, at pH 7.0, both proteins are dimers with an extended content of beta-sheet conformation, but pH 2.0 and 9.0 yield a different secondary, tertiary and quaternary structural organisation. Whilst beta-LgA is a monomer, that conserves beta-sheet conformation at pH 2.0 and 9.0, GdA has a stable dimeric structure at alkaline pH, but at pH 2.0 increases its alpha-helix content and it aggregates soon. SR beam has been used to perform SAXS comparative measurements of the two proteins. SR-SAXS data provide the radius of gyration and the radii of the cross section and of the thickness. GdA aggregation at acid pH has been characterised by calculating the distance distribution function (P(r)). Isoelectric focusing and chromatofocusing data show a different charge distribution on the surfaces of the two proteins, supporting the hypothesis that the presence of oligosaccharides deeply influences the conformational state and the aggregation process of GdA at different pH values. In particular, the presence of sialic acid residues, within the oligosaccharide moiety of the GdA, might be responsible for the differences observed between the two proteins. PMID- 11004544 TI - Purification, physico-chemical characterization and sequence of a heat labile alkaline metalloprotease isolated from a psychrophilic Pseudomonas species. AB - The psychrophilic alkaline metalloprotease (PAP) produced by a Pseudomonas bacterium isolated from Antarctica has been purified and characterized. The gene encoding PAP has been cloned and sequenced and the derived amino acid sequence shows 66% identity with the mesophilic alkaline metalloprotease from Pseudomonas aeruginosa IFO 3455 (AP). Compared to the purified AP, PAP is three times more active at 20 degrees C, is very sensitive to chelating agents and is rapidly inactivated at 45 degrees C. The lower thermostability of PAP can tentatively be explained by a loss of a stabilizing Ca(2+), a decrease in the content of hydrophobic residues and a smaller aliphatic index. PMID- 11004545 TI - Conformational study of a custom antibacterial peptide cecropin B1: implications of the lytic activity. AB - Cecropin B1 (CB1) with two amphipathic alpha-helical segments is a derivative of the natural antibacterial peptide, cecropin B. The assays of cell lysis show that, compared with cecropin A (CA), CB1 has a similar ability to lyse bacteria with a higher potency (two- to six-fold higher) in killing cancer cells. The difference may be due to the fact that the peptides possess different structures and sequences. In this study, the solution structure of CB1 in 20% hexafluoroisopropanol was determined by two-dimensional nuclear magnetic resonance (NMR) spectroscopy. The (1)H NMR resonances were assigned. A total of 350 inter-proton distances were used to calculate the solution structure of CB1. The final ensemble structures were well converged, showing the minimum root mean square deviation. The results indicate that CB1 has two stretches of helices spanning from residues 3 to 22 and from residues 26 to 33, which are connected by a hinge section formed by Gly-23 and Pro-24. Lys-25 is partially incorporated in the hinge region. The bent angle between two helical segments located in two planes was between 100 and 110 degrees. With comparisons of the known NMR structure of CA and its activities on bacteria and cancer cells, the structure function relationship of the peptides is discussed. PMID- 11004546 TI - Stoichiometry and inter-subunit interaction of the wedge initiation complex, gp10 gp11, of bacteriophage T4. AB - Association of gp10 and gp11 (gp=gene product) is the first step in the assembly pathway of the wedge part of the baseplate of bacteriophage T4. The gp10-gp11 complex constitutes the six tail pins at the corners of the baseplate hexagon on the distal side. The stoichiometry of the subunits, gp10 and gp11, of this complex was determined in combination with sedimentation equilibrium, Edman degradation of the complex and sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). From the results of Edman degradation and SDS-PAGE, the molar ratio of gp10 and gp11 was approximately 1. On the other hand, the molecular weight of the purified gp10-gp11 complex was determined by sedimentation equilibrium to be 284000+/-7000, which is in good agreement with the expected value of 269840 if the stoichiometry is 3:3. Furthermore, comparison of the results in the presence and in the absence of reducing reagent, 2 mercaptoethanol (2-ME), in SDS-PAGE revealed that two molecules of gp10 in the complex formed a disulfide bond, while the third gp10 molecule does not participate in the disulfide bond formation. PMID- 11004547 TI - A catalytic site of protein disulfide isomerase probed with adenosine-5' triphosphate analogs. AB - Anthraniloyl adenosine-5'-triphosphate (Ant-ATP) and etheno-adenosine-5' triphosphate (epsilon-ATP) complexed to Mg(2+) ions are substrates of protein disulfide isomerase (PDI). epsilon-ATP, coordinated to Tb(3+) ions, was used as a probe of the ATPase binding site. Sensitized luminescence arising from resonance energy transfer from epsilon-adenine to Tb(3+) is quenched by PDI. The luminescence results are discussed in reference to a model in which the distance of separation between epsilon-adenine (donor) and Tb(3+) (acceptor) is increased upon binding of PDI. The interaction of a small peptide of 14 amino acid residues with the b/b' domain of the protein does not influence the ATPase activity. The phosphorescence, fluorescence and fluorescence anisotropy of bound epsilon-ATP are not perturbed by the binding of the small molecular weight peptide to PDI. It is suggested that the peptide and ATP do not share a common binding site on the b/b' domain. PMID- 11004548 TI - Quantitative characterization of homo- and heteroassociations of muscle phosphofructokinase with aldolase. AB - Dissociation of purified phosphofructokinase accompanied with inactivation was analyzed in the absence and presence of aldolase and the data were compared with those obtained with muscle extract. The kinetics of the decrease in enzymatic activity was highly dependent on the dilution factor in both cases, but the inactivation appeared to be biphasic only with extract. The inactivation of the phosphofructokinase was impeded by addition of excess of aldolase. Time courses of kinase inactivation were fitted by alternative kinetic models to characterize the multiple equilibria of several homo- and hetero-oligomers of phosphofructokinase. The combination of modeling data obtained with purified and extract systems suggests that aldolase binds to an intermediate dimer of phosphofructokinase and within this heterocomplex the kinase is completely active. The intermediate dimer is stabilized by association with microtubules and the kinase activity decreased due to dilution can be recovered by addition of excess aldolase. In extract, the phosphofructokinase is of sigmoidal character (Hill coefficient of 2.3); the addition of excess exogenous aldolase to phosphofructokinase resulted in heterocomplex formation displaying Michaelian kinetics. The possible physiological relevance of heterocomplex formation of phosphofructokinase in muscle extract is discussed. PMID- 11004549 TI - The adhesive protein of Choromytilus chorus (Molina, 1782) and Aulacomya ater (Molina, 1782): a proline-rich and a glycine-rich polyphenolic protein. AB - The adhesive polyphenolic proteins from Aulacomya ater and Choromytilus chorus with apparent molecular masses of 135000 and 105000, respectively, were digested with trypsin and the peptides produced resolved by reversed phase liquid chromatography. About 5 and 12 major peptides were obtained from the protein of A. ater and C. chorus, respectively. The major peptides were purified by reverse phase chromatography and the amino acid sequence indicates that both polyphenolic proteins consisted of repeated sequence motifs in their primary structure. The major peptides of A. ater contain seven amino acids corresponding to the consensus sequence AGYGGXK, whereas the tyrosine was always found as 3, 4 dihydroxyphenylalanine (Dopa), the X residue in position 6 was either valine, leucine or isoleucine, and the carboxy terminal was either lysine or hydroxylysine. On the other hand, the major peptides of C. chorus ranged in size from 6 to 21 amino acids and the majority correspond to the consensus sequence AKPSKYPTGYKPPVK. Both proteins differ markedly in the sequence of their tryptic peptides, but they share the common characteristics of other adhesive proteins in having a tandem sequence repeat in their primary structure. PMID- 11004550 TI - Thermochemistry of the specific binding of C12 surfactants to bovine serum albumin. AB - The specific binding to bovine serum albumin (BSA) of anionic and non-ionic surfactants with C12 acyl chains has been studied by high sensitivity isothermal titration calorimetry. This method proved particularly effective in resolving the binding of anionic surfactants into separate classes of sites with different affinity. For sodium dodecylsulfate (SDS) the measured binding curves could be rationalized as association to two classes (high affinity/low affinity) of sites comprising, respectively, three and six similar (i.e. thermodynamically equivalent), independent sites. Changes in the thermodynamic functions enthalpy, standard free energy, standard entropy and heat capacity could be discerned for each class of binding site, as well as for micelle formation. These data suggest that binding to low affinity sites (in analogy with micelle formation) exhibits energetic parameters; in particular, a large negative change in heat capacity, which is characteristic of hydrophobic interactions. The thermodynamics of high affinity binding, on the other hand, is indicative of other dominant forces; most likely electrostatic interactions. Other anionic ligands investigated (laurate and dodecyl benzylsulfonate) showed a behavior similar to SDS, the most significant difference being the high affinity binding of the alkylbenzyl sulfonate. For this ligand, the thermodynamic data is indicative of a more loosely associated complex than for SDS and laurate. BSA was found to bind one or two of the non-ionic surfactants (NIS) hepta- or penta(ethylene glycol) monododecyl ether (C12EO7 and C12EO5) with binding constants about three orders of magnitude lower than for SDS. Hence, the free energy of the surfactant in the weakly bound BSA-NIS complex is only slightly favored over the micellar state. The binding process is characterized by very large exothermic enthalpy changes (larger than for the charged surfactants) and a large, positive increment in heat capacity. These observations cannot be reconciled with a molecular picture based on simple hydrophobic condensation onto non-polar patches on the protein surface. PMID- 11004551 TI - Effect of P(2)' site tryptophan and P(20)' site deletion of Momordica charantia trypsin inhibitor II on inhibition of proteinases. AB - Momordica charantia trypsin inhibitor II (MCTI-II) inhibits the amidolytic activity of factor Xa with a K(i) value 10-100-fold smaller than those of other squash family inhibitors. It also inhibits factor X activation mediated by factor VIIa-tissue factor complex or factor IXa. Comparison of other squash family inhibitors reveal Trp at position 7 (P(2)') and a deletion at position 25 (P(20)') are characteristics of MCTI-II. In order to elucidate the effect of these positions on the inhibitory activity, we chemically synthesized three inhibitors: S-MCTI-II whose amino acid sequence is identical to natural MCTI-II, S-MCTI-II(7L) whose P(2)'(Trp) is substituted with Leu, and S-MCTI-II(25N) whose P(20)'(deletion) is filled with Asn. The dissociation constants of the complexes of human factor Xa with S-MCTI-II, S-MCTI-II(7L), and S-MCTI-II(25N) were 1.3x10( 6) M, 2.8x10(-5) M, and 7.3x10(-6) M, respectively. They inhibited factor X activation mediated by factor VIIa with the same degree. As in the case of natural MCTI-II, S-MCTI-II suppressed factor X activation mediated by factor IXa, while S-MCTI-II(7L) and S-MCTI-II(25N) did not. Both the Trp at the P(2)' position and deletion at the P(20)' position are thus likely required for the inhibition of factor Xa, trypsin, and factor IXa, while these two positions do not affect factor X activation initiated by the factor VIIa-tissue factor complex. PMID- 11004552 TI - pH-induced domain interaction and conformational transitions of lipoxygenase-1. AB - The multidomain structure of soybean LOX1 was examined over the pH range 1-12. Lipoxygenase-1 activity was reversible over broad pH range of 4-10 due to the reversibility of conformational states of the molecule. Below pH 4.0, due to collapse in hydrophobic interactions, the enzyme unfolded to an irreversible conformation with the properties of molten globule state with a mid point of transition at pH 2.4. This intermediate state lost iron irreversibly. In alkaline pH at 11.5, LOX1 underwent partial unfolding with the exposure of cysteine residues with subsequent oxidation of a pair of cysteine residues in the C terminal domain and this intermediate showed some properties of molten globule state and retained 35% of activity. Beyond pH 12.0, the enzyme was completely inactivated irreversibly due to irreversible conformational changes. The pH dependent urea-induced unfolding of LOX1 suggested that LOX1 was more stable at pH 7.0 and least stable at pH 9.0. Furthermore, the urea-induced unfolding of LOX1 indicated that the unfolding was biphasic due to pH-dependent domain interactions and involved sequential unfolding of domains. The loss of enzyme activity at pH 4. 0 and 7.0 occurred much earlier to unfolding of the C-domain at all pHs studied. The combination of urea-induced unfolding measurements and limited proteolysis experiments suggested that at pH 4.0, the domains in LOX1 were less interactive and existed as tightly folded units. Furthermore, these results confirmed the contribution of ionic interactions in the interdomain contacts. PMID- 11004553 TI - Arginine to citrulline replacement in substrates of phosphorylase kinase. AB - Synthetic peptides based on residues 9 to 18 of glycogen phosphorylase were prepared containing citrulline at position 10 or 16, or at both positions 10 and 16. The peptides were compared as substrates for a recombinant, truncated form of the catalytic subunit of phosphorylase kinase (residues 1-300). The peptide having citrulline at position 10 was phosphorylated the same as the parent peptide. Both the peptides with a single citrulline at position 16 and with two citrullines were phosphorylated less effectively than the parent peptide; k(cat)/K(m) values were approximately 20% the value with the parent peptide. Incorporation of the second citrulline had little change in the effectiveness of the peptide as a substrate although the kinetic parameters with the citrulline peptides did show differences. The change in peptide phosphorylation did not seem to result from a change in peptide structure. Two-dimensional nuclear magnetic resonance studies of di-citrulline peptide are reported and showed no change in the solution structure of the peptide compared to the parent peptide. Thus, the change in kinetic parameters with the modified peptides seemed an effect of arginine replacement and was likely a consequence of the loss of charge inasmuch as the size of arginine and citrulline are similar. Arginine-16 was concluded to be more important for phosphorylase kinase recognition than arginine-10. These findings were consistent with the earlier studies using alanine replacement of arginine in synthetic peptides as substrates for the holoenzyme form of phosphorylase kinase. PMID- 11004554 TI - Structure and function of alpha-fetoprotein: a biophysical overview. AB - alpha-Fetoprotein (AFP) is a large serum glycoprotein belonging to the intriguing class of onco-developmental proteins. AFP has attracted considerable attention since it was shown that the change in its serum level during pregnancy is a hallmark of the development of numerous embryonic disorders, while the increase in its content in the plasma of adults correlates with the appearance of several pathological conditions. Over the past 30 years, some 11000 papers have been published concerning AFP, an average rate of over a publication a day since 1969. The majority of publications are about the application of the protein in diagnostics, or about other uses of AFP in biomedicine; though some of them describe the biochemical and functional properties of AFP, two aspects have been extensively reviewed. However, surprisingly little is currently known about structural properties of this protein as well as about the molecular mechanism of its function. The present review pursues the aim to describe the current state of the art in studies of structural properties and conformational stability of AFP. An attempt to establish the relationship between conformational transformations in AFP and its function is also made. PMID- 11004555 TI - Structural changes in cytochrome c oxidase induced by cytochrome c binding. A resonance raman study. AB - Electrostatically stabilized complexes of fully oxidized cytochrome c oxidase from Paracoccus denitrificans and horse heart cytochrome c were studied by resonance Raman spectroscopy. The experiments were carried out with the wild-type oxidase and a variant in which a negatively charged amino acid in the binding domain (D257) is replaced by an asparagine. It is shown that cytochrome c induces structural changes at heme a and heme a(3) which are reminiscent to those found in mammalian cytochrome c oxidase-cytochrome c complex. The spectral changes are attributed to subtle changes in the heme-protein interactions implying that there is a structural communication from the binding domain even to the remote catalytic center. Only for the heme a modes minor spectral differences were found in the response of the wild-type and the D257N variant oxidase upon cytochrome c binding indicating that electrostatic interactions of aspartate 257 are not crucial for the perturbation of the catalytic site structure in the complex. On the other hand, in none of the complexes, structural changes were detected in the bound cytochrome c. These findings are in contrast to previous results obtained with beef heart cytochrome c oxidase which triggers the formation of a new conformational state of cytochrome c assumed to be involved in the biological electron transfer process. PMID- 11004556 TI - Characterisation of two protein phosphatase 2A holoenzymes from maize seedlings. AB - Two holoenzymes of protein phosphatase 2A (PP2A), designated PP2AI and PP2AII, were purified from maize seedlings. The subunit composition of maize holoenzymes generally resembled those of animal PP2A. Using SDS/PAGE and Western blots with antibodies generated against peptides derived from animal PP2A, we established the subunit composition of plant protein phosphatase 2A. In both maize holoenzymes, a 38000 catalytic (PP2Ac) and a 66000 constant regulatory subunit (A) constituting the core dimer of PP2A were present. In addition, PP2AI (180000 200000) contained a protein of 57000 which reacted with antibodies generated against the peptide (EFDYLKSLEIEE) conserved in all eukaryotic Balpha regulatory subunits. In contrast, none of the proteins visualised in PP2AII (140000-160000) by double staining reacted with these antibodies. The activity of PP2AI measured with (32)P-labelled phosphorylase a in the presence of protamine and ammonium sulfate is about two times higher than that of PP2AII. PP2AI and PP2AII displayed different patterns of activation by protamine, polylysine and histone H1 and exhibit high sensitivity toward inhibition by okadaic acid. The data obtained provide direct biochemical evidence for the existence in plants of PP2A holoenzymes composed of a catalytic subunit complexed with one or two regulatory subunits. PMID- 11004557 TI - Is cellobiose dehydrogenase from Phanerochaete chrysosporium a lignin degrading enzyme? AB - Cellobiose dehydrogenase (CDH) is an extracellular redox enzyme of ping-pong type, i.e. it has separate oxidative and reductive half reactions. Several wood degrading fungi produce CDH, but the biological function of the enzyme is not known with certainty. It can, however, indirectly generate hydroxyl radicals by reducing Fe(3+) to Fe(2+) and O2 to H2O2. Hydroxyl radicals are then generated by a Fenton type reaction and they can react with various wood compounds, including lignin. In this work we study the effect of CDH on a non-phenolic lignin model compound (3,4-dimethoxyphenyl glycol). The results indicate that CDH can affect lignins in three important ways. (1) It breaks beta-ethers; (2) it demethoxylates aromatic structures in lignins; (3) it introduces hydroxyl groups in non-phenolic lignins. The gamma-irradiated model compound gave a similar pattern of products as the CDH treated model compound, when the samples were analyzed by HPLC, suggesting that hydroxyl radicals are the active component of the CDH system. PMID- 11004558 TI - Effects of ionic surfactants used in reversed micelles on cutinase activity and stability. AB - The effects of aqueous surfactant solutions on the kinetics and stability of cutinase from Fusarium solani pisi were studied. The surfactant sodium bis[2 ethylhexyl]ester sulfosuccinic acid (AOT) acts as a pseudo-competitive inhibitor within a limited concentration range relative to the hydrolysis of short-chain p nitrophenyl esters. For higher concentrations a hyperbolic mixed inhibition takes place. A pseudo-activation of hydrolysis in presence of AOT and hexadecyltrimethyl-ammonium bromide (CTAB) was observed. CTAB has similar effects on kinetics of cutinase. Cutinase revealed to be stable in CTAB solutions, with activity retention as high as 80%. AOT has a deleterious effect on the enzyme in the time course, resulting in acute loss of activity possibly related with unfolding of the protein structure. A relation between deactivation rate constants and AOT/cutinase concentration ratios is suggested. The presence of the linear alcohol, 1-hexanol, was included in these solutions, in the attempt to interpret the deactivation of cutinase when encapsulated in reversed micelle systems in the absence of this co-surfactant. PMID- 11004559 TI - Enzymic characteristics of secreted aspartic proteases of Candida albicans. AB - Candida yeasts are rarely infectious, but frequently cause life-threatening systemic infections in patients immunocompromised by AIDS or by immunosuppressive therapeutics. The secreted aspartic proteases (Saps) are known virulence factors of pernicious Candida species. The most virulent, Candida albicans, possesses at least nine SAP genes, some of which are specifically expressed from cells with morphologies associated with virulence. Only one of these proteases, Sap2, has been previously purified from yeast in sufficient quantities for enzymic studies. The other enzymes are present in low amounts in yeast culture and are difficult to purify. As a consequence, enzyme properties, including the substrate specificities, of all Saps are poorly studied. Therefore, four Saps that are known to be expressed in C. albicans, Sap1, Sap2, Sap3 and Sap6, were produced in Escherichia coli as recombinant zymogens and purified in large quantities. These proenzymes were autoactivated and purified as active proteases. The enzymic properties including the substrate specificities at the P(1) and P(1)' sites were determined using a competitive hydrolysis method employing synthetic substrate mixtures. All four Saps cleave peptide bonds between larger hydrophobic amino acids, but these somewhat broad specificities differ in detail among the four enzymes at both sites. At the P(1) site, Sap1, Sap2 and Sap6 prefer Phe while Sap3 prefers Leu. Positively charged amino acids are also accommodated, especially by Sap2 and Sap3. The specificities at P(1)' are broader than at P(1) for all four enzymes. Sap6 prefers Ala, whereas other Saps prefer Tyr. Acidic side chains are also accommodated at this site. Analysis of substrates with a hydrophobic amino acid in P(1)' reveals that all the Saps possess a unique preference for Ala at this site. The observed differences of residue preferences among Saps may be utilized for the design of specific substrates and inhibitors. PMID- 11004560 TI - Specificity of DNA binding and methylation by the M.FokI DNA methyltransferase. AB - The M.FokI adenine-N(6) DNA methyltransferase recognizes the asymmetric DNA sequence GGATG/CATCC. It consists of two domains each containing all motifs characteristic for adenine-N(6) DNA methyltransferases. We have studied the specificity of DNA-methylation by both domains using 27 hemimethylated oligonucleotide substrates containing recognition sites which differ in one or two base pairs from GGATG or CATCC. The N-terminal domain of M.FokI interacts very specifically with GGATG-sequences, because only one of the altered sites is modified. In contrast, the C-terminal domain shows lower specificity. It prefers CATCC-sequences but only two of the 12 star sites (i.e. sites that differ in 1 bp from the recognition site) are not accepted and some star sites are modified with rates reduced only 2-3-fold. In addition, GGATGC- and CGATGC-sites are modified which differ at two positions from CATCC. DNA binding experiments show that the N terminal domain preferentially binds to hemimethylated GGATG/C(m)ATCC sequences whereas the C-terminal domain binds to DNA with higher affinity but without specificity. Protein-protein interaction assays show that both domains of M.FokI are in contact with each other. However, several DNA-binding experiments demonstrate that DNA-binding of both domains is mutually exclusive in full-length M.FokI and both domains do not functionally influence each other. The implications of these results on the molecular evolution of type IIS restriction/modification systems are discussed. PMID- 11004561 TI - Characterization of yeast seryl-tRNA synthetase active site mutants with improved discrimination against substrate analogues. AB - The involvement of amino acids within the motif 2 loop of Saccharomyces cerevisiae seryl-tRNA synthetase (SerRS) in serine and ATP binding was demonstrated previously [B. Lenhard et al., J. Biol. Chem. 272 (1997) 1136-1141]. In our attempt to analyze the structural basis for the substrate specificity and to explore further the catalytic mechanism employed by S. cerevisiae SerRS, two new active site mutants, SerRS11 and SerRS12, were constructed. The catalytic effects of amino acid replacement at positions Lys287, Asp288 and Ala289 with purified wild-type and mutant seryl-tRNA synthetases were tested. The alteration of these semi-conserved amino acids interferes with tRNA-dependent optimization of serine recognition. Additionally, mutated enzymes SerRS11 (Lys287Thr, Asp288Tyr, Ala289Val) and SerRS12 (Lys287Arg) are less sensitive to inhibition by two competitive inhibitors: serine hydroxamate, an analogue of serine, and 5'-O [N-(L-seryl)-sulfamoyl]adenosine, a stable analogue of aminoacyl adenylate, than the wild-type enzyme. SerRS mutants also display different activation kinetics for serine and serine hydroxamate, indicating that specificity toward the substrates is modulated by amino acid replacement in the motif 2 loop. PMID- 11004562 TI - The K(+) affinity of gastric H(+),K(+)-ATPase is affected by both lipid composition and the beta-subunit. AB - It is generally assumed that negatively charged residues present in the alpha subunit of gastric H(+),K(+)-ATPase are involved in K(+) binding and transport. Despite the fact that there is no difference between various species regarding these negatively charged residues, it was observed that the apparent K(+) affinity of the pig enzyme was much lower than that of the rat H(+),K(+)-ATPase. By determining the K(+)-stimulated dephosphorylation reaction of the phosphorylated intermediate K(0.5) values for K(+) of 0.12+/-0.01 and 1.73+/-0.03 mM were obtained (ratio 14.4) for the rat and the pig enzyme, respectively. To investigate the reason for the observed difference in K(+) sensitivity, both enzymes originating from the gastric mucosa were either reconstituted in a similar lipid environment or expressed in Sf9 cells. After reconstitution in K(+) permeable phosphatidylcholine/cholesterol liposomes K(0.5) values for K(+) of 0.16+/-0.01 and 0.35+/-0.05 mM for the rat and pig enzyme respectively were measured (ratio 2.2). After expression in Sf9 cells the pig gastric H(+),K(+) ATPase still showed a 4.1 times lower K(+) sensitivity than that of the rat enzyme. This means that the difference in K(+) sensitivity of the rat and pig gastric H(+), K(+)-ATPase is not only due to a different lipid composition but also to the structure of either the alpha- or beta-subunit. Expression of hybrid enzymes in Sf9 cells showed that the difference in K(+) sensitivity between the rat and pig gastric H(+),K(+)-ATPase is primarily due to differences in the beta subunit. PMID- 11004563 TI - Modifications of aclacinomycin T by aclacinomycin methyl esterase (RdmC) and aclacinomycin-10-hydroxylase (RdmB) from Streptomyces purpurascens. AB - The genes rdmB and rdmC of Streptomyces purpurascens encoding aclacinomycin modifying enzymes RdmB and RdmC were expressed in Streptomyces lividans TK24. In contrast to the earlier suggestion that RdmC may be an esterase that causes the removal of the carbomethoxy group from the 10 position of aclacinomycins, RdmC functions as an aclacinomycin methyl esterase and catalyzes the removal of the methoxy group from the C-15 position of aclacinomycin T producing 15 demethoxyaclacinomycin T. RdmB acts upon C-10 of 15-demethoxyaclacinomycin T and is able to remove the carboxylic group from the C-10 position. It functions also as an aclacinomycin-10-hydroxylase being able to add a hydroxyl group at the same, C-10 position in vitro. Aclacinomycin methyl esterase was purified to apparent homogeneity from S. lividans carrying the rdmC and aclacinomycin-10 hydroxylase as a glutathione S-transferase fusion construct from Escherichia coli carrying the rdmB gene, respectively. Aclacinomycin methyl esterase functions as a monomer and aclacinomycin-10-hydroxylase as a tetramer. Aclacinomycin methyl esterase has an exceptionally high temperature stability and has an apparent K(m) for aclacinomycin T of 15.5 microM. The introduction of rdmC and rdmB in a Streptomyces galilaeus mutant HO38 produced the same modifications of aclacinomycin T in vivo as aclacinomycin methyl esterase and aclacinomycin-10 hydroxylase in vitro. PMID- 11004564 TI - Trichloroacetic acid and trifluoroacetic acid-induced unfolding of cytochrome c: stabilization of a native-like folded intermediate(1). AB - A systematic investigation of trichloroacetic acid (TCA) and trifluoroacetic acid (TFA)-induced equilibrium unfolding of native horse cytochrome c has been carried out using a combination of optical spectroscopy and electrospray ionization mass spectroscopy (ESI MS). In the presence of an increasing concentration of TCA the native cytochrome c does not undergo significant unfolding but stabilization of a partially folded intermediate is observed. This TCA-induced partially folding intermediate of cytochrome c had an enhanced secondary structure and slightly disrupted tertiary structure compared to native protein and undergoes extensive unfolding in the presence of TFA. However, in the presence of an increasing concentration of TFA, cytochrome c was found to undergo extensive unfolding characterized by a significant breakdown of the secondary and tertiary structure of protein. The TFA-unfolded cytochrome c was found to undergo folding in the presence of TCA and low guanidine hydrochloride (GdmCl) resulting in the stabilization of the partially folded intermediate. The effectiveness of TCA as compared to TFA in the stabilization of intermediates was further supported by the observation that low concentrations of TCA were found to induce refolding of HCl-denatured cytochrome c whereas, under similar concentrations of acid, no significant effect on the unfolded structure of protein was observed in the presence of TFA. ESI MS studies indicated that the trichloroacetate anion has a greater affinity for cytochrome c compared to trifluoroacetate anion, which might be the reason for the stabilization of the native-like folded intermediate during TCA-induced denaturation of cytochrome c as compared to extensive unfolding observed in the presence of TFA. PMID- 11004565 TI - Colloidal properties of human transferrin receptor in detergent free solution. AB - The colloidal properties of transferrin receptor, isolated from human placenta, in detergent free solution has been investigated by light scattering techniques and analytical ultracentrifugation. In detergent free solution at 293.2 K, hTfR forms stable aggregates with an apparent hydrodynamic radius of 17 nm. The molecular mass was determined by ultracentrifugation to lie between (1722+/-87) kDa (sedimentation equilibrium) and (1675+/-46) kDa (sedimentation velocity). This implies that the aggregates are build up from nine hTfR dimers. Based on model calculations, which are in good agreement with the experimental data, we propose a torus-like structure for the aggregates. Upon pH shift from pH 7.5 to 5.0 or removal of the N-linked carbohydrate chains, formation of larger aggregates is induced. These aggregates can be described in terms of porous fractal structures. We propose a simple model, which accounts for that behaviour assuming that the aggregation is mainly due to the reduction of negative surface charge. PMID- 11004566 TI - Active site studies of bovine alpha1-->3-galactosyltransferase and its secondary structure prediction. AB - The catalytic domain of bovine alpha1-->3-galactosyltransferase (alpha3GalT), residues 80-368, have been cloned and expressed, in Escherichia coli. Using a sequential purification protocol involving a Ni(2+) affinity column followed by a UDP-hexanolamine affinity column, we have obtained a pure and active protein from the soluble fraction which catalyzes the transfer of galactose (Gal) from UDP-Gal to N-acetyllactosamine (LacNAc) with a specific activity of 0.69 pmol/min/ng. The secondary structural content of alpha3GalT protein was analyzed by Fourier transform infrared (FTIR) spectroscopy, which shows that the enzyme has about 35% beta-sheet and 22% alpha-helix. This predicted secondary structure content by FTIR spectroscopy was used in the protein sequence analysis algorithm, developed by the Biomolecular Engineering Research Center at Boston University and Tasc Inc., for the assignment of secondary structural elements to the amino acid sequence of alpha3GalT. The enzyme appears to have three major and three minor helices and five sheet-like structures. The studies on the acceptor substrate specificity of the enzyme, alpha3GalT, show that in addition to LacNAc, which is the natural substrate, the enzyme accepts various other disaccharides as substrates such as lactose and Gal derivatives, beta-O-methylgalactose and beta-D thiogalactopyranoside, albeit with lower specific activities. There is an absolute requirement for Gal to be at the non-reducing end of the acceptor molecule which has to be beta1-->4-linked to a second residue that can be more diverse in structure. The kinetic parameters for four acceptor molecules were determined. Lactose binds and functions in a similar way as LacNAc. However, beta O-methylgalactose and Gal do not bind as tightly as LacNAc or lactose, as their K(ia) and K(A) values indicate, suggesting that the second monosaccharide is critical for holding the acceptor molecule in place. The 2' and 4' hydroxyl groups of the receiving Gal moiety are important in binding. Even though there is large structural variability associated with the second residue of the acceptor molecule, there are constraints which do not allow certain Gal-R sugars to be good acceptors for the enzyme. The beta1-->4-linked residue at the second position of the acceptor molecule is preferred, but the interactions between the enzyme and the second residue are likely to be non-specific. PMID- 11004567 TI - Phosphohexose isomerase/autocrine motility factor/neuroleukin/maturation factor is a multifunctional phosphoprotein. AB - Phosphohexose isomerase (PHI) is a member of the ectoenzyme/exoenzyme family and plays a key role in both glycolysis and gluconeogenesis pathways. Upon secretion PHI acts as a cytokine with tumor autocrine motility factor (AMF), neuroleukin (NLK) and maturation factor (MF) functions. Signaling is initiated by its binding to a cell surface 78 kDa glycoprotein (gp78). However, since PHI protein is a 'leaderless' secretory protein, released from cells via a non-classical route(s), we questioned whether the molecule undergoes post-translation modification while retaining proper folding and maintaining intact enzymatic and motogenic activities. To address this, we have generated, expressed and isolated a recombinant human AMF (rhAMF). The rhAMF retained the biological activities of the native AMF, i.e., catalyzes phosphohexose isomerization and stimulated cell motility. Additionally, we show here that human PHI is phosphorylated at serine 185 by casein kinase II (CK II) and we provide experimental evidence suggesting that this phosphorylation is associated with secretion, thus providing insights for elucidating the intracellular signal transmission of cell response to stimulation by AMF/NLK/MF. PMID- 11004568 TI - Diverse mechanisms of inhibition of pyruvate dehydrogenase kinase by structurally distinct inhibitors. AB - The mechanism of action of structurally distinct pyruvate dehydrogenase kinase (PDK) inhibitors was examined in assays with experimental contexts ranging from an intact pyruvate dehydrogenase complex (PDC) with and without supplemental ATP or ADP to a synthetic peptide substrate to PDK autophosphorylation. Some compounds directly inhibited the catalytic activity of PDKs. Some of the inhibitor classes tested inhibited autophosphorylation of recombinant PDK1 and PDK2. During these studies, PDC was shown to be directly inhibited by a novel mechanism; the addition of supplemental recombinant PDKs, an effect that is ADP dependent and partly alleviated by members of each of the compound classes tested. Overall, these data demonstrate that small molecules acting at diverse sites can inhibit PDK activity. PMID- 11004569 TI - Structure-function studies on Taiwan cobra long neurotoxin homolog. AB - A novel long neurotoxin homolog was purified from Naja naja atra (Taiwan cobra) venom using the combination of ion exchange chromatography and reverse phase high performance liquid chromatography. The determined protein sequence was essentially the same as that deduced from the cDNA amplified by reverse transcriptase-polymerase chain reaction. The long neurotoxin homolog exhibited an activity that inhibited acetylcholine-induced muscle contractions, as with N. naja atra cobrotoxin. The degree of inhibition caused by the addition of long neurotoxin homolog was approximately 70% of that observed with the addition of cobrotoxin. Unlike the well-known short and long neurotoxins, this neurotoxin homolog contained two additional cysteine residues forming a disulfide linkage in the N-terminal region. Circular dichroism measurement and computer models of the neurotoxin reveal that its secondary structure was not abundant in beta-sheet as noted with short and long neurotoxins. This less ordered structure may be associated with the lower activity noted with the long neurotoxin homolog. Together with the finding that the known long neurotoxin homologs exclusively appear in the venoms of the Naja and Bungarus genera, the long neurotoxin homologs should represent an evolutionary branch from the long and short neurotoxins in the Elapidae family. PMID- 11004570 TI - Mechanism of the inhibition of milk xanthine oxidase activity by metal ions: a transient kinetic study. AB - The nature and mechanism of the inhibition of the oxidoreductase activity of milk xanthine oxidase (XO) by Cu(2+), Hg(2+) and Ag(+) ions has been studied by steady state and stopped flow transient kinetic measurements. The results show that the nature of the inhibition is noncompetitive. The inhibition constants for Cu(2+) and Hg(2+) are in the micromolar and that for Ag(+) is in the nanomolar range. This suggests that the metal ions have strong affinity towards XO. pH dependence studies of the inhibition indicate that at least two ionisable groups of XO are involved in the binding of these metal ions. The effect of the interaction of the metal ions on the reductive and oxidative half reactions of XO has been investigated, and it is observed that the kinetic parameters of the reductive half reaction are not affected by these metal ions. However, the interaction of these metal ions with XO significantly affects the kinetic parameters of the oxidative half reaction. It is suggested that this may be the main cause for the inhibition of XO activity by the metal ions. PMID- 11004571 TI - Characterisation of a homogeneous plant aminoaldehyde dehydrogenase. AB - According to our knowledge, this is the first purification method developed, enabling isolation of a homogeneous aminoaldehyde dehydrogenase (AMADH) from etiolated pea seedlings. The procedure involved initial purification with precipitants followed by three low pressure chromatographic steps. Partially purified enzyme was further subjected to fast protein liquid chromatography on a Mono Q column and to affinity-interaction chromatography on 5'-AMP Sepharose. Purity of the final enzyme preparation was checked by sodium dodecyl sulfate polyacrylamide gel electrophoresis and chromatofocusing. Pea AMADH exists as a tetramer of 230 kDa in the native state, a molecular mass of one subunit was determined as 57 kDa. The enzyme was found to be an acidic protein with pI 5.4. AMADH showed a broad substrate specificity utilising various aminoaldehydes (C3 C6) as substrates. The best substrate of pea AMADH was 3-aminopropionaldehyde, the enzyme also efficiently oxidised 4-aminobutyraldehyde and omega guanidinoanalogues of the aminoaldehydes. Pea AMADH was inhibited by SH reagents, several elementary aldehydes and metal-binding agents. Although AMADH did not oxidise betaine aldehyde at all, the N-terminal amino acid sequence of the enzyme shows a high degree of homology with those of plant betaine aldehyde dehydrogenases (BADHs) of spinach, sugar beet and amaranth. Several conserved amino acids were found in comparison with BADH from cod liver of known crystal structure. PMID- 11004572 TI - The role of tryptophan residues in substrate binding to catalytic domains A and B of xylanase C from Fibrobacter succinogenes S85. AB - Oxidation of the isolated catalytic domain B of xylanase C (XynC-B) from Fibrobacter succinogenes with N-bromosuccinimide (NBS) resulted in the modification of five of the seven Trp residues present in the enzyme. Hydrolytic activity of the enzyme was rapidly lost upon initiation of oxidation as a molar ratio of about two NBS molecules per molar equivalent of protein was sufficient to cause 50% inhibition of enzyme activity, and the addition of five molar equivalents of NBS resulted in less than 10% activity. Pre-incubation of XynC-B with the competitive inhibitor D-xylose resulted in the apparent protection of two Trp residues from oxidation. Xylose protection of the enzyme also resulted in a maintenance of activity, with 60% activity still evident after addition of 8-9 molar equivalents of NBS. This protection from inactivation was enhanced by the inclusion of xylohexaose in reaction mixtures. Under these conditions, however, a further Trp residue was protected from NBS oxidation. The three protected Trp residues were identified as Trp135, Trp161 and Trp202 by differential labelling and peptide mapping of NBS-oxidized preparations of the xylanase employing a combination of electrospray mass spectroscopic analysis and N-terminal sequencing. By analogy to the known structures of the family 11 xylanases, the fully conserved Trp202 residue is located on the only alpha-helix present in the enzymes, at the interface between it and the back of the beta-sheet which forms the active site cleft. Trp135 represents a highly conserved aromatic residue in family 11, but it is replaced with Thr in domain A of F. succinogenes xylanase C. To investigate the role of Trp135 in conferring the different activity profile of domain B relative to domain A, the Trp135Thr and Trp135Ala derivatives of domain B were prepared by site-directed mutagenesis. However, the kinetic parameters of the two domain B derivatives were not significantly different compared to the wild-type enzyme as reflected by K(M) and k(cat) values and product distribution profiles. Similar results were obtained with the Trp161Ala derivative of domain B, indicating that these two residues do not directly participate in the binding of substrate but likely form the foundation for binding subsite 2. PMID- 11004573 TI - High-pressure denaturation of apomyoglobin. AB - The pressure denaturation of wild type and mutant apomyoglobin (apoMb) was investigated using a high-pressure, high-resolution nuclear magnetic resonance and high-pressure fluorescence techniques. Wild type apoMb is resistant to pressures up to 80 MPa, and denatures to a high-pressure intermediate, I(p), between 80 and 200 MPa. A further increase of pressure to 500 MPa results in denaturation of the intermediate. The two tryptophans, both in the A helix, remain sequestered from solvent in the high-pressure intermediate, which retains some native NOESY cross peaks in the AGH core as well as between F33 and F43. High-pressure fluorescence shows that the tryptophans remain inaccessible to solvent in the I(p) state. Thus the high-pressure intermediate has some structural properties in common with the apoMb I(2) acid intermediate. The resistance of the AGH core to pressures up to 200 MPa provides further evidence that the intrinsic stability of these alpha-helices is responsible for their presence in a number of equilibrium intermediates as well as in the earliest kinetic folding intermediate. Mutations in the AGH core designed to disrupt packing by burying a charge or increasing the size of a hydrophobic residue significantly perturbed the unfolding of native apoMb to the high-pressure intermediate. The F123W and S108L mutants both unfolded at lower pressures, while retaining some resistance to pressures below 50 MPa. The charge burial mutants, A130K and S108K, are not stable at very low pressures and both denature to the intermediate by 100 MPa, half of the pressure required for wild type apoMb. Thus a similar intermediate state is created independent of the method of perturbation, and mutations have similar effects on native state destabilization for both methods of denaturation. These data suggest that equilibrium intermediates that can be formed through different means are likely to resemble a kinetic intermediate. PMID- 11004574 TI - Crystallization and preliminary X-ray crystallographic analysis of NADPH: azodicarbonyl/quinone oxidoreductase, a plant zeta-crystallin. AB - Arabidopsis thaliana P1 protein was crystallized using the hanging drop vapor diffusion method in 0.1 M piperazine-1, 4-bis(2-ethanesulfonic acid) buffer, containing 14% polyethylene glycol 6000 and 0.2 M magnesium acetate at pH 6.5 and 20 degrees C. The crystals are orthorhombic and belong to the space group P2(1)2(1)2(1) with unit cell dimensions of a=49.8, b=122.4 and c=149. 9 A. The diffraction data up to 2.9 A were collected by a multiwire area detector. PMID- 11004575 TI - Purification, crystallisation and preliminary X-ray study of haemoglobin from Crocodilis palustris and Crocodilis porosus. AB - The unsolved three-dimensional structure of crocodile haemoglobin and its prospects as a blood substitute have led us to initiate the purification and crystallisation of haemoglobin molecules from crocodile species (Crocodilis palustris or mugger and Crocodilis porosus or salt water crocodile). The work has resulted in the prevention of polymerisation of naked haemoglobin molecules using N-ethylmaleimide or iodoacetamide. The purified monomeric haemoglobin molecule of C. porosus was crystallised in two different forms and X-ray diffraction data were collected up to 2 A resolution for both forms. Form I: a=53.62, b=53.55, c=103.77 A; beta=93.35 degrees, space group P2(1), Z=2. Form II: a=71.30, b=54.70, c=80.00 A; beta=106.4 degrees, space group P2(1), Z=2. Structure solution and rigid body refinement of form I data resulted in a model with R(free)=0.42 and R=0.35. PMID- 11004576 TI - Escherichia coli is able to produce heterologous tetraheme cytochrome c(3) when the ccm genes are co-expressed. AB - The production of Desulfovibrio vulgaris Hildenborough cytochrome c(3) (M(r) 13000), which is a tetraheme cytochrome, in Escherichia coli was examined. This cytochrome was successfully produced in an E. coli strain co-expressing the ccmABCDEFGH genes involved in the cytochrome c maturation process. The apocytochrome c(3) was matured in either anaerobic or aerobic conditions, but aerobic growth in the presence of delta-aminolevulinic acid was found to be best for cytochrome c(3) production. Site-directed mutagenesis was performed to investigate the effect of the presence of four amino acids in between the two cysteines of the heme binding sites 2 and 4 on the maturation of holocytochrome c(3) in E. coli. PMID- 11004577 TI - Contributions of protein disulfide isomerase domains to its chaperone activity. AB - Protein disulfide isomerase (PDI), a member of the thioredoxin (Trx) superfamily, consists of five consecutive domains, a-b-b'-a'-c. Domain combinations, AB, A'C, B'A'C and AB-C, and hybrids of PDI domains with Trx, Trx-C and Trx-B'A'C, have been constructed and expressed in Escherichia coli to examine the contributions of PDI domains to its enzyme and chaperone activities. All the combination and hybrid products are considerably less active than intact PDI in their enzyme activities. Recombinant products containing C, at low concentrations, inhibit the reactivation of lysozyme in HEPES buffer, while those without C do not. Only the intact PDI molecule and the hybrid molecule, Trx-B'A'C, but to a much lower level, show general chaperone activity in assisting the reactivation of denatured D-glyceraldehyde-3-phosphate dehydrogenase. It is suggested that all domains of PDI contribute to the binding of target protein for its chaperone activity. PMID- 11004578 TI - Enthalpic destabilization of glycosylated lysozymes constructed by genetic modification. AB - To understand the role of polyglycosylation in protein stability, the thermodynamic changes in the denaturation of various polymannosyl lysozyme mutants (R21T, G49N, R21T/G49N) constructed by genetic modification were analyzed using differential scanning calorimetry (DSC). The denaturation temperature and the enthalpy change for unfolding of the lysozymes were reduced with an increase in the length of the polymannose chain and the number of binding sites to a protein, although the polymannosyl lysozymes revealed apparent heat stability in that no aggregation was observed and the enzymatic activity was conserved under conditions in which the wild-type lysozyme coagulated [S. Nakamura et al., J. Biol. Chem. 268 (1993) 12706-12712]. The reversibility of the denaturation of polymannosyl lysozymes was observed in the DSC curves obtained by reheating after heat denaturation, while it was not observed for the wild-type lysozyme. Based on these results, the polymannosyl lysozyme seems to easily refold due to the excellent reversibility of denaturation, despite the decreases in the enthalpic stabilization due to the strain in the protein molecule by the introduction of a polysaccharide chain. PMID- 11004579 TI - The native metastable fold of C1-inhibitor is stabilized by disulfide bonds. AB - C1-inhibitor is a member of the serpin family of proteinase inhibitors and is an important inhibitor of complement and contact system proteinases. The native protein has the characteristic serpin feature of being in a kinetically trapped metastable state rather than in the most stable state it could adopt. A consequence of this is that it readily forms loop-sheet dimers and polymers, by a mechanism believed to be the same as observed with other serpins. An unusual feature of C1-inhibitor is that it has a unique amino-terminal domain, of unknown function, held to the serpin domain by two disulfide bonds not found in other serpins. We report here that reduction of these bonds by DTT, causes a conformational change such that the reactive center loop inserts into beta-sheet A. This form of C1-inhibitor is less stable to heat and urea than the native protein, and is more susceptible to extensive degradation by trypsin. These data show that the disulfide bonds in C1-inhibitor are required for the protein to be stabilized in the metastable state with the reactive center loop expelled from beta-sheet A. PMID- 11004580 TI - Specific interaction of lipoate at the active site of rhodanese. AB - Dihydrolipoate is an acceptor of the rhodanese-bound sulfane sulfur atom, as shown by analysis of the elementary steps of the reaction catalyzed by rhodanese. The crystal structure of sulfur-substituted rhodanese complexed with the non reactive oxidized form of lipoate has revealed that the compound is bound at the enzyme active site, with the dithiolane ring buried in the interior of the cavity and the carboxylic end pointing towards the solvent. One of the sulfur atoms of the ligand in the unproductive complex is relatively close to the sulfane sulfur bound to Cys-247, the sulfur that is transferred during the catalytic reaction. This mode of binding of lipoate is likely to mimic that of dihydrolipoate. The results presented here support the possible role of dihydrolipoate as sulfur acceptor substrate of rhodanese in an enzymatic reaction that might serve to provide iron-sulfur proteins with inorganic sulfide. PMID- 11004581 TI - Comparison of HOCl traps with myeloperoxidase inhibitors in prevention of low density lipoprotein oxidation. AB - In this study, the production of the highly toxic oxidant hypochlorous acid (HOCl) by the phagocytic enzyme myeloperoxidase (MPO) was quantitated and the concomitant alterations of low density lipoprotein (LDL) were analyzed in view of the potential role of LDL in atherosclerosis. Using the monochlorodimedone assay, it was found that HOCl is produced in micromolar concentrations. The kinetics of the decrease of tryptophan fluorescence appeared to be a sensitive method to monitor LDL alterations under near in vivo conditions. Therefore, this method was used to subsequently compare the effectiveness of MPO inhibitors that block production of HOCl with compounds that act as HOCl traps. The efficiency of MPO inhibitors to prevent LDL damage increased in the series benzohydroxamic acid < salicylhydroxamic acid < 3-amino-1,2,4-triazole < sodium azide < potassium cyanide < p-hydroxy-benzoic acid hydrazide, while for the HOCl traps the protective efficiency increased in the series glycine < taurine < methionine. We conclude that HOCl traps may have high potential therapeutic impact in vivo due to their low toxicity, although high concentrations of them would have to reach sites of inflammation. In contrast, only low concentrations of a specific MPO inhibitor would be required to irreversibly inhibit the enzyme. PMID- 11004582 TI - Characterization of a heme c nitrite reductase from a non-ammonifying microorganism, Desulfovibrio vulgaris Hildenborough. AB - A cytochrome c nitrite reductase (NiR) was purified for the first time from a microorganism not capable of growing on nitrate, the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. It was isolated from the membranes as a large heterooligomeric complex of 760 kDa, containing two cytochrome c subunits of 56 and 18 kDa. This complex has nitrite and sulfite reductase activities of 685 micromol NH(4)(+)/min/mg and 1.0 micromol H(2)/min/mg. The enzyme was studied by UV-visible and electron paramagnetic resonance (EPR) spectroscopies. The overall redox behavior was determined through a visible redox titration. The data were analyzed with a set of four redox transitions, with an E(0)' of +160 mV (12% of total absorption), -5 mV (38% of total absorption), -110 mV (38% of total absorption) and -210 mV (12% of total absorption) at pH 7.6. The EPR spectra of oxidized and partially reduced NiR show a complex pattern, indicative of multiple heme-heme magnetic interactions. It was found that D. vulgaris Hildenborough is not capable of using nitrite as a terminal electron acceptor. These results indicate that in this organism the NiR is not involved in the dissimilative reduction of nitrite, as is the case with the other similar enzymes isolated so far. The possible role of this enzyme in the detoxification of nitrite and/or in the reduction of sulfite is discussed. PMID- 11004583 TI - Incorporation of nitrotyrosine into alpha-tubulin by recombinant mammalian tubulin-tyrosine ligase. AB - Tubulin-tyrosine ligase (TTL, EC 6.3.2.25) from porcine brain, which catalyses the readdition of tyrosine to the C-terminus of detyrosinated alpha-tubulin, was cloned and expressed in Escherichia coli as a glutathione S-transferase-fusion protein. Upon cleavage of the immobilised fusion protein, an electrophoretically homogeneous enzyme was obtained. Recombinant TTL, which exhibited similar catalytic properties as the mammalian enzyme purified from brain tissue, was capable of using nitrotyrosine as an alternative substrate in vitro. Incorporation of tyrosine into tubulin was competitively inhibited by nitrotyrosine with an apparent K(i) of 0.24 mM. The TTL-catalysed incorporation of nitrotyrosine as sole substrate into alpha-tubulin was clearly detectable at concentrations of 10 microM by immunological methods using nitrotyrosine specific antibodies. However, in competition with tyrosine 20-fold higher concentrations of nitrotyrosine were necessary before its incorporation became evident. Analysis of the C-terminal peptides of in vitro modified alpha-tubulin by MALDI-MS confirmed the covalent incorporation of nitrotyrosine into tubulin by TTL. In contrast to the C-terminal tyrosine, pancreatic carboxypeptidase A was incapable of cleaving nitrotyrosine from the modified alpha-tubulin. PMID- 11004584 TI - Eosin interaction of alpha-synuclein leading to protein self-oligomerization. AB - Among various dyes including congo red, thioflavin S, thioflavin T, eosin, rhodamine 6G, and phenol red, the eosin was the only dye that induced self oligomerization of alpha-synuclein in the presence of a chemical coupling reagent of N-(ethoxycarbonyl)-2-ethoxy-1, 2-dihydroquinoline. To analyze chemical nature of the eosin interaction with alpha-synuclein, the phenomenon of self oligomerization was further examined with eosin congeners such as ethyl eosin, eosin B, phloxine B, erythrosin B, and rose bengal. The followings are the conclusions we have reached. First of all, intactness of the benzoate moiety of eosin and the negative charge on the carboxylic group of the dye are important factors leading to the specific interaction with alpha-synuclein. Secondly, the localized negative charge on the xanthene moiety of eosin is another critical factor for the interaction. As far as substituting halides are concerned, bromides and iodides on the xanthene moiety of the dyes do not make any difference on the alpha-synuclein interaction because both eosin and erythrosin B have induced the common phenomenon of self-oligomerization. The binding curve between eosin and alpha-synuclein was sigmoidal as the dye concentrations were increased. A double reciprocal plot of the saturation curve showed that the maximum number of eosin binding sites on alpha-synuclein was 1.85 with a dissociation constant of 390 microM. The dye binding to the protein appeared to occur via a positive cooperativity. The eosin binding site(s) was suggested to be located predominantly on the NAC region and partly related to the acidic C terminus of alpha-synuclein. It has been, therefore, expected that this information might be useful to develop alpha-synuclein interactive molecules, which could provide eventual preventive or possible therapeutic means against various alpha-synuclein related disorders including Parkinson's disease. PMID- 11004585 TI - STIM1: a novel phosphoprotein located at the cell surface. AB - STIM1 is a novel candidate growth suppressor gene mapping to the human chromosome region 11p15.5 that is associated with several malignancies. STIM1 overexpression studies in G401 rhabdoid tumour, rhabdomyosarcoma and rodent myoblast cell lines causes growth arrest, consistent with a potential role as a tumour growth suppressor. We used highly specific antibodies to show by immunofluorescence and cell surface biotinylation studies that STIM1 is located at the cell surface of K562 cells. Western blot analysis revealed that the 90-kDa STIM1 protein is ubiquitously expressed in various human primary cells and tumour cell lines. STIM1 is not secreted from cells and does not appear to undergo proteolytic processing. We show evidence of post-translational modification of STIM1, namely phosphorylation and N-linked glycosylation. Phosphorylation of STIM1 in vivo occurs predominantly on serine residues. Thus, STIM1, the putative tumour growth suppressor gene is ubiquitously expressed and has features of a regulatory cell surface phosphoprotein. PMID- 11004605 TI - Adapting to multiple personalities: Cbl is also a RING finger ubiquitin ligase. PMID- 11004606 TI - A growing role for Rho family GTPases as intermediaries in growth factor- and adhesion-dependent cell cycle progression. PMID- 11004607 TI - Fish oil diet affects on oxidative senescence of red blood cells linked to degeneration of spleen cells in mice. AB - The effect of dietary polyunsaturated fatty acids and alpha-tocopherol supplementation on erythrocyte lipid peroxidation and immunocompetent cells in mice was studied comparatively using seven dietary oils (15% oil/diet, w/w) including fish oil rich in eicosapentaenoic acid (EPA, 20:5, n-3) and docosahexaenoic acid (DHA, 22:6, n-3). A 43% increase in spleen weight, about twice as many spleen cells and no change in the subpopulations of spleen cells, as well as a significant depression of mitogen-induced blastogenesis of both T and B cells in the spleen were observed in mice fed fish oil for 30 days in comparison with soybean oil diet-fed mice. In the fish oil diet-fed mice, membranous lipid hydroperoxide (hydroperoxides of phosphatidylcholine and phosphatidylethanolamine) accumulation as a marker of oxidative senescence in red blood cells (RBC) was 2.7-3.5 times higher than that in mice fed soybean oil, although there was no difference in the plasma phosphatidylcholine hydroperoxide concentration. In spite of the supplementation of alpha-tocopherol to up to 10 times the level in the basal diet, the degeneration of spleen cells and the stimulated oxidative senescence of RBC found by the fish oil feeding could not be prevented. The results suggest that oral intake of excess polyunsaturated fatty acids, i.e. EPA and DHA, in a fish oil diet can lead to acceleration of membrane lipid peroxidation resulting in RBC senescence linked to the lowering of immune response of spleen cells, and that supplementation of alpha-tocopherol as antioxidant does not always effectively prevent such oxidative degeneration as observed in spleen cells and RBC in vivo. PMID- 11004608 TI - 1H-NMR lipid profiles of human blood platelets; links with coronary artery disease. AB - Blood platelets are closely involved in the early development of atherosclerosis and in the events that lead to thrombosis, both of which are dominating factors in coronary artery disease (CAD). The aim of the present study was to evaluate the platelet lipid profiles of patients suffering from CAD and explore the possibility of a link between platelet lipids and CAD, using high-resolution high field proton nuclear magnetic resonance spectroscopy as the analytical tool. The total platelet lipid profiles of healthy volunteers were compared with those of patients presenting with chest pain requiring coronary angiography. Two lipid groups changed significantly: cholesterol increased by 16.5% and total diacylglycerophospholipids decreased by 15.7%. There was also a significant decrease of the ethanolamine-containing phospholipids, by 4.7%; the extent of unsaturation of the fatty acid chains, by 0.2, and increase of the linoleate content of the fatty acid chains, by 1.9%. Our results suggest that platelet lipid abnormalities occur in patients with CAD and these changes may predate the development of overt atherosclerosis. PMID- 11004609 TI - Arachidonic acid-containing phosphatidylcholine inhibits lymphocyte proliferation and decreases interleukin-2 and interferon-gamma production from concanavalin A stimulated rat lymphocytes. AB - The proliferation of concanavalin A (Con A)-stimulated rat lymphocytes was markedly inhibited by phosphatidylcholine containing arachidonic and stearic acids (PC(A-S)), but not by phosphatidylcholine containing oleic and stearic acids or phosphatidylinositol containing arachidonic and stearic acids. The concentration of PC(A-S) which inhibited Con A-stimulated proliferation by 50% was 31 microM and near total inhibition was observed at 154 microM . Phosphatidylserine containing only oleic acid enhanced proliferation by 37% at a concentration of 31 microM , but phosphatidylethanolamine and phosphatidylcholine containing only oleic acid did not affect proliferation at this concentration. It is concluded that both the head group and the fatty acid composition contribute to the influence of phospholipids on lymphocyte proliferation. The effects of PC(A-S) on T-lymphocyte responses were investigated further. In parallel with the inhibition of proliferation PC(A-S) caused a concentration-dependent decrease in the production of the Th1-type cytokines interleukin (IL)-2 and interferon (IFN) gamma; inhibition of cytokine production was >85% at the highest concentration of PC(A-S) used (154 microM ). Production of the Th2-type cytokines IL-4 and IL-10 was not affected. The possible role of prostaglandins in mediating the effects of PC(A-S) was examined by adding indomethacin into the medium and the participation of lipid peroxidation was examined by adding vitamin E and vitamin C. Indomethacin and vitamin E did not affect the inhibition caused by PC(A-S) but vitamin C caused a partial reversal. It is concluded that inhibition of T lymphocyte proliferation by phospholipids involves both the head group and the fatty acyl chains, that this inhibition is not mediated by prostaglandins but may involve some form of oxidant stress and that some phospholipids (e.g., PC(A-S)) can markedly influence cytokine profiles. PMID- 11004610 TI - Assay of microsomal oxysterol 7alpha-hydroxylase activity in the hamster liver by a sensitive method: in vitro modulation by oxysterols. AB - A method of assaying hepatic cytochrome P-450, oxysterol 7alpha-hydroxylase (CYP7B), was developed by combining the use of 25-[26,27-(3)H]hydroxycholesterol as a substrate and hydroxypropyl-beta-cyclodextrin as a substrate vehicle. When these assay conditions were tested, an undesirable transformation was observed of the reaction product, 7alpha,25-dihydroxycholesterol, into 3-oxo-7alpha,25 dihydroxy-4-cholesten by the activity of 3beta-hydroxy-Delta(5)-C(27) steroid oxydoreductase, a microsomal NAD(+) and NADP(+) dependent enzyme of bile acid metabolism. A great improvement was reached by using a continuous NADPH generating system which constantly re-transforms NADP(+) into NADPH, thus inhibiting this activity. This improved CYP7B assay, comparable to our previously described assay for cholesterol 7alpha-hydroxylase (CYP7A), allowed a 3-fold increase of the apparent enzyme activity. The possibility to simultaneously measure CYP7A and CYP7B activities on the same microsomal preparation was investigated. A marked decrease (-33%) in the CYP7B activity was noticed, while that of CYP7A remained unchanged. The CYP7B activity was observed to be inhibited by cholesterol (-30%) and also by the oxysterols 7alpha-hydroxycholesterol ( 21%), 7beta-hydroxycholesterol (-25%) and epicoprostanol (-20%), and by cyclosporin A (-26%). It can be concluded that this sensible and easy to perform CYP7B assay allows to observe, at least in vitro, a modulation of the enzyme activity by oxysterols. PMID- 11004611 TI - Cloning and promoter analysis of the cotton lipid transfer protein gene Ltp3(1). AB - A cotton Ltp3 gene and its 5' and 3' flanking regions have been cloned with a PCR based genomic DNA walking method. The amplified 2.6 kb DNA fragment contains sequences corresponding to GH3 cDNA which has been shown to encode a lipid transfer protein (LTP3). The gene has an intron of 80 bp which is located in the region corresponding to the C-terminus of LTP3. The Ltp3 promoter was systematically analyzed in transgenic tobacco plants by employing the Escherichia coli beta-glucuronidase gene (GUS) as a reporter. The results of histochemical and fluorogenic GUS assays indicate that the 5' flanking region of the Ltp3 gene contains cis-elements conferring the trichome specific activity of Ltp3 promoter. PMID- 11004612 TI - Cicatricial pemphigoid. AB - Cicatricial pemphigoid (CP) is a heterogeneous group of rare, chronic, subepithelial blistering disorders of the mucous membranes and, occasionally, the skin, which can have serious and rarely fatal consequences. The most common clinical features are desquamative gingivitis, oral erosions, and conjunctival fibrosis. Skin lesions occur less frequently and may present as widespread vesicles and bullae, as in bullous pemphigoid (BP). In some patients, the scarring can be a source of significant morbidity because it can result in odynophagia, strictures of the upper aerodigestive tract, or corneal opacities leading to eventual blindness. This article is a comprehensive review and discusses clinical, pathologic, and pathophysiologic aspects of this group of disorders collectively known as CP. (J Am Acad Dermatol 2000;43:571-91.) LEARNING OBJECTIVE: At the conclusion of this learning activity, participants should be familiar with the clinical spectrum of CP, the histopathologic and immunopathologic characteristics, the differential diagnosis, the treatment, and the natural history of the disease. Furthermore, this learning activity should facilitate early diagnosis of CP and should promote the idea that the involvement of other specialists, including ophthalmologists, otolaryngologists, gastroenterologists, and oral medicine specialists, as appropriate, will aid in providing these patients with the highest quality of care. PMID- 11004613 TI - Anti-CD4 monoclonal antibody treatment of moderate to severe psoriasis vulgaris: results of a pilot, multicenter, multiple-dose, placebo-controlled study. AB - BACKGROUND: OKTcdr4a (IMUCLONE) is a humanized anti-CD4 IgG4 monoclonal antibody that retains the binding and in vitro immunosuppressive properties of the parent murine antibody. Psoriasis is a chronic disease for which treatment with multiple doses of monoclonal antibodies is likely to be required for adequate control. OBJECTIVE: This study was performed to test the efficacy and safety of OKTcdr4a, given in sequential courses over a period of several weeks, in the treatment of moderate to severe psoriasis vulgaris. METHODS: Twenty-eight patients (45.6 +/- 10.1 years of age) were studied, with a mean pretreatment Psoriasis Area and Severity Index (PASI) score of 18.3. In the first double-blind phase of the study, patients were randomized to receive OKTcdr4a as a 225 mg/course (low dose), 750 mg/course (high dose), or placebo divided into 3 identical infusions over a 5-day period. After 42 days, patients who met the criteria for re treatment with OKTcdr4a were re-treated with the 750 mg/course in an open phase of the study. RESULTS: After the double-blind course of treatment, the mean PASI decreased by 11% in the placebo group, by 4% in the low-dose group, and by 17% in the high-dose group at 15 days. Twenty patients met the criteria for re-treatment (ie, did not experience a decrease in PASI score of 50% at 42 days). They were re treated with OKTcdr4a at 43 days with the 750 mg/course in the open phase of the study. By day 99, the mean PASI score decreased from 19.9 at baseline to 17 in those patients who had received either placebo or low-dose OKTcdr4a followed by high-dose OKTcdr4a. In contrast, the mean PASI score decreased from 17.4 at baseline to only 7.7 in those patients who had received high-dose OKTcdr4a for both courses. Sustained CD4 saturation was not necessary for sustained clinical response. No patients had significant changes in circulating CD4(+) T-cell counts. The infusions were well tolerated. CONCLUSION: Targeting CD4 using sequential treatments with a humanized monoclonal antibody (OKTcdr4a) may offer another therapeutic option for the treatment of moderate to severe psoriasis. PMID- 11004614 TI - Late metastases of cutaneous melanoma: an analysis of 31 patients. AB - Analysis of data of 6298 patients with cutaneous melanoma seen at the Department of Dermatology and Allergology at the Ludwig-Maximilians University of Munich between 1977 and 1998 identified 31 patients who first experienced metastatic disease 10 or more years after surgical treatment of the primary melanoma. The mean tumor thickness in patients with late progression of disease was 1.4 mm. There was no sex predominance. Of 31 patients, 15 (48. 3%) were female and included both premenopausal and postmenopausal women at the time of initial diagnosis without any predominance. Sixteen of the 31 patients (51.6%) with late onset of progression had primary tumors located on the trunk. The prognosis after relapse was related to the site of recurrence; survival after local or regional node recurrence was often prolonged, whereas survival after distant metastases usually was limited. Survival after late recurrence did not correlate with disease-free survival. A high-risk group for late recurrence could not be identified. The data emphasize the value of continued clinical follow-up and self examination for early detection of regional recurrences. We advocate a lifelong follow-up, also to identify other melanocytic lesions at risk (eg, dysplastic nevi, secondary or multiple melanomas). PMID- 11004616 TI - Sensitivity and specificity of clinical, histologic, and immunologic features in the diagnosis of paraneoplastic pemphigus. AB - BACKGROUND: Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease characterized by the production of autoantibodies mainly directed against proteins of the plakin family. An overlapping distribution of autoantibody specificities has been recently reported between PNP, pemphigus vulgaris (PV), and pemphigus foliaceus (PF), which suggests a relationship between the different types of pemphigus. OBJECTIVE: Our purpose was to evaluate the sensitivity and the specificity of clinical, histologic, and immunologic features in the diagnosis of PNP. METHODS: The clinical, histologic, and immunologic features of 22 PNP patients were retrospectively reviewed and compared with those of 81 PV and PF patients without neoplasia and of 8 PV and 4 PF patients with various neoplasms. RESULTS: One clinical and 2 biologic features had both high sensitivity (82%-86%) and high specificity (83%-100%) whatever the control group considered: (1) association with a lymphoproliferative disorder, (2) indirect immunofluorescence (IIF) labeling of rat bladder, and (3) recognition of the envoplakin and/or periplakin bands in immunoblotting. Two clinicopathologic and two biologic features had high specificity (87%-100%) but poor sensitivity (27% 59%): (1) clinical presentation associating erosive oral lesions with erythema multiforme-like, bullous pemphigoid-like, or lichen planus-like cutaneous lesions; (2) histologic picture of suprabasal acantholysis with keratinocyte necrosis, interface changes, or lichenoid infiltrate; (3) presence of both anti epithelial cell surface and anti-basement membrane zone antibodies by IIF; and (4) recognition of the desmoplakin I and/or BPAG1 bands in immunoblotting. Interestingly, 45% of patients with PNP presented initially with isolated oral erosions that were undistinguishable from those seen in PV patients, and 27% had histologic findings of only suprabasal acantholysis, which was in accordance with the frequent detection of anti-desmoglein 3 antibodies in PNP sera. CONCLUSION: The association with a lymphoproliferative disorder, the IIF labeling of rat bladder, and the immunoblotting recognition of envoplakin and/or periplakin are both sensitive and specific features in the diagnosis of PNP. PMID- 11004615 TI - Effect of daily versus intermittent sunscreen application on solar simulated UV radiation-induced skin response in humans. AB - BACKGROUND: Acute and chronic skin damage occurs as a consequence of solar UV radiation exposure. To diminish such skin damage, the dermatologic community advocates the daily use of sunscreens as part of a sun avoidance strategy. OBJECTIVE: We determined the effectiveness of a sunscreen product with a sunscreen protection factor (SPF) of 15 applied daily in preventing UV-induced histologic damage in human skin compared with the protection afforded by sunscreens with equal or higher SPF applied intermittently. METHODS: Twenty-four subjects were exposed to 2 minimal erythema doses of solar-simulated UV on 4 consecutive days. Three sunscreen products were applied to the buttock of each subject. One SPF 15 product was applied daily before exposure to UV and, to simulate intermittent product use, an SPF 15 or SPF 29 product was applied on 3 of 4 days, with one missed application on days 2, 3, or 4. Skin biopsy specimens were taken and processed for routine and immunohistochemical staining. Changes in number of sunburn cells and Langerhans cells as well as degree of inflammatory infiltrate and lysozyme immunostaining were determined. RESULTS: There was a statistically significant increase in the number of sunburn cells, degree of inflammation, and intensity of lysozyme staining, and there was a decrease in the number of Langerhans cells at sites where sunscreen application was missed as compared with unirradiated control and daily SPF 15 sunscreen-treated sites. CONCLUSION: Our data suggest that daily use of a sunscreen reduces the skin damage produced by UV exposure compared with intermittent use of equal or higher SPF products. The daily application of sunscreens in appropriate quantities reduces the harmful effects of solar UV radiation on skin. Compliance is essential for maximal benefit of sunscreens. PMID- 11004617 TI - Prognostic indicators in venous ulcers. AB - BACKGROUND: Venous ulcers can be difficult to heal, and prognostic factors for healing have not been fully elucidated. OBJECTIVE: The objective of this study was to analyze the results of a large multicenter venous ulcer trial to retrospectively establish prognostic factors for venous ulcer healing. METHODS: This study examined data from a previously published prospective randomized placebo-controlled trial of an oral medication versus placebo treatment for venous ulcers. Local leg ulcer care involved the use of a moisture-retentive dressing and sustained graduated compression with a paste bandage and a self adherent wrap. The oral medication or placebo was administered on a daily basis with the same dressings and bandage system in both groups for 12 weeks. A total of 165 patients completed the full 12-week treatment period; 83 received ifetroban, 82 received placebo. RESULTS: There was no statistically significant difference in outcome between the two groups. The study showed that consistent local ulcer treatment with a clearly defined system of care was associated with an unexpectedly high percentage (55%) of long-standing large venous ulcers (mean duration, 27 months; mean area, 15.9 cm(2)) being healed in both groups. Baseline ulcer area and duration of leg ulcer were found to be important in predicting outcome. Ulcers of short duration were found to be most likely to heal. Percent healing and ulcer area at week 3 were good predictors of 100% healing. Ulcers that had at least 40% healing by week 3 predicted more than 70% of the outcomes correctly. CONCLUSION: From this large study it was determined that baseline ulcer area and ulcer duration are significant predictors of 100% healing and time to heal. Percent healing and ulcer area at week 3 are good predictors of complete ulcer healing. Ulcers that are large, long-standing, and slow to heal after 3 weeks of optimal therapy are unlikely to heal rapidly, and might benefit from alternative therapies. PMID- 11004618 TI - Raynaud's phenomenon, anticentromere antibodies, and digital necrosis without sclerodactyly: an entity independent of scleroderma? AB - We describe 4 women of 43, 73, 84, and 86 years with Raynaud's phenomenon, severe digital necrosis, and high serum levels of anticentromere antibodies without skin thickening or internal organ sclerosis. Investigations revealed no diabetes or arterial vascular disease leading to arterial obstruction. Histologic examination did not show any dermal sclerosis or calcinosis. The intravenous infusions of prostaglandin reversed the ischemic lesions in 3 patients. These cases suggest that the triad Raynaud's phenomenon, anticentromere antibodies and necrosis of digits without sclerodactyly and sclerosis of internal organ should be considered as an entity distinct from scleroderma with sclerosis. For this entity we propose the name RACAND syndrome. PMID- 11004619 TI - Rapidly progressing mycosis fungoides presenting as follicular mucinosis. AB - Follicular mucinosis can occur as a primary idiopathic disorder or can arise in association with benign or malignant disease, most notably mycosis fungoides. We describe a patient with an aggressive folliculotropic variant of mycosis fungoides that initially presented as follicular mucinosis with alopecia. One month after the diagnosis of follicular mucinosis, a diagnosis of mycosis fungoides was made, and 3 months later inguinal lymph node involvement with mycosis fungoides developed. A skin biopsy specimen demonstrated prominent follicular mucinosis with folliculotropism of atypical cells and intrafollicular Pautrier's microabscesses. As demonstrated in this case, follicular mucinosis can be a presenting sign of rapidly progressive mycosis fungoides. In our review of follicular mucinosis and its association with mycosis fungoides, we found that the folliculotropic variant of mycosis fungoides appears more commonly to have an aggressive course than classic mycosis fungoides. PMID- 11004620 TI - A large-scale North American study of fungal isolates from nails: the frequency of onychomycosis, fungal distribution, and antifungal susceptibility patterns. AB - BACKGROUND: Onychomycosis, a fungal infection of the nail bed, is responsible for up to 50% of nail disorders. Although several surveys have been conducted in different parts of the world, there have been no multicenter epidemiologic surveys of onychomycosis in North America. OBJECTIVE: A 12-center study was undertaken to (1) determine the frequency of onychomycosis, (2) identify organisms recovered from the nails, and (3) determine the antifungal susceptibility of isolates. METHODS: A total of 1832 subjects participated in this study and completed a comprehensive questionnaire, and nail clippings were collected for potassium hydroxide examination and culturing. RESULTS: The frequency of onychomycosis, as defined by the presence of septate hyphae on direct microscopy and/or the recovery of a dermatophyte, was found to be 13.8%. In general, the dermatophyte isolates were susceptible to the antifungals tested. CONCLUSION: Because of the limited number of large-scale studies, the baseline incidence is not firmly established. However, the higher frequency of onychomycosis in this study may confirm the suspected increase in incidence of disease in North America. PMID- 11004621 TI - The prevalence of atopic dermatitis in Oregon schoolchildren. AB - BACKGROUND: Although surveys from many parts of the world have shown that the prevalence of atopic dermatitis (AD) in schoolchildren has increased greatly in the past 40 years, there is no current prevalence information from the United States. OBJECTIVE: Our objective was to investigate the utility of a recently developed European questionnaire to estimate the prevalence of AD in urban and rural Oregon schoolchildren. METHODS: The self-administered Schultz-Larsen questionnaire (SLQ) of AD symptoms and history was completed by the parents of a broad socioeconomic and ethnic mix of 5- to 9-year-old schoolchildren from 6 urban and 2 rural elementary schools in Oregon. Validation assessments included comparisons of the questionnaire scores with clinical examination in a group of age-matched children and with specific history components known to correlate with AD. RESULTS: Data showed a prevalence of 17.2% using standard scoring criteria for the SLQ and with a lower limit of 6.8% according to highly stringent criteria derived from the validation study using dermatologic examination. A single question ("Has a doctor ever said that your child has eczema?") was highly concordant with the questionnaire determination, yielding very high predictive accuracy (91.2%). CONCLUSION: This study of childhood AD frequency indicates a high prevalence of AD in the United States, comparable to that recently observed from studies in Europe and Japan. (J Am Acad Dermatol 2000;43:649-55.). PMID- 11004622 TI - Photodamage pilot study: a double-blind, vehicle-controlled study to assess the efficacy and safety of tazarotene 0.1% gel. AB - BACKGROUND: Tazarotene, a potent acetylenic retinoid for topical use, might be expected to benefit photodamaged skin, including improving the classical signs of fine wrinkles, mottled hyperpigmentation, and roughness. OBJECTIVE: Our purpose was to determine the efficacy and safety of tazarotene 0.1% gel in the treatment of photodamaged dorsal forearm skin. METHODS: Ten healthy female volunteers, aged 45 to 65 years, with moderately photodamaged forearm skin applied tazarotene 0.1% gel to one arm and vehicle gel to the other once daily for 12 weeks. The study was a double-blind, randomized, paired-comparison evaluation conducted at a single site. RESULTS: Tazarotene showed beneficial effects for several efficacy variables. It was more efficacious than vehicle in reducing skin roughness and fine wrinkling based on objective measurements. Tazarotene also corrected epidermal atrophy and atypia and improved skin hydration properties. CONCLUSION: In this 12-week pilot study tazarotene redressed abnormalities associated with photo-damaged skin. PMID- 11004623 TI - Basal cell carcinoma in chronic arsenicism occurring in Queensland, Australia, after ingestion of an asthma medication. AB - BACKGROUND: Ingestion of trivalent inorganic arsenic has long been recognized as a cause of basal cell carcinomas (BCCs) and has been reported most often in Taiwan and Singapore. OBJECTIVE: Our purpose was to study the clinical and histologic characteristics of BCCs occurring in Australian Caucasians as a consequence of chronic arsenicism due to ingestion of an arsenic-containing medication. METHODS: Self-referred persons with a history of ingestion of Bell's Asthma Medication were interviewed, and skin examinations were performed. Local age- and sex-matched patients with BCCs were used to compare the distribution and histologic subtypes of BCCs in arsenic-exposed and sporadic cases. RESULTS: Thirty-six persons (21 male, 15 female; mean age, 57 years) participated, all of whom had been exposed to the asthma medication early in life (mean age, 13 years) for a mean duration of 5 years. Each person had at least one cutaneous sign of chronic arsenicism, either self-reported or on examination, and all except one had a history of either BCC or squamous cell carcinoma of the skin, with self reports of 20 to 2000 skin lesions removed per person. The mean age at first presentation with a BCC was 33 years, but neither latency nor number of skin lesions appeared to be related to duration of exposure to arsenic. BCCs in persons exposed to arsenic occurred more often on sun-protected sites compared with BCCs in age- and sex-matched sporadic cases (P <.001), but the distribution and histologic subtypes between these two groups were similar. CONCLUSION: We have described BCCs in arsenic-exposed Australians and shown that they occur predominantly in sun-protected locations. Although the reported number of skin lesions is very high, the latency and number do not appear to be related to the duration of arsenic exposure. The histologic types of the BCCs occurring in arsenic-exposed persons are not different from sporadic BCCs. PMID- 11004624 TI - Ultraviolet A1 (340-400 nm) phototherapy for scleroderma in systemic sclerosis. AB - BACKGROUND: The presence of an inflammatory infiltrate consisting of helper T cells and a dysregulated matrix metabolism leading to excessive deposition of collagen are two pathogenetic factors responsible for the developments of fibrosis and sclerosis in patients with systemic sclerosis. In previous studies, ultraviolet A1 (UVA1) radiation phototherapy was shown to deplete skin infiltrating T cells through the induction of T-cell apoptosis and to up-regulate the expression of matrixmetalloproteinase-1 (collagenase-1) in dermal fibroblasts. OBJECTIVE: Our purpose was to determine whether UVA1 phototherapy is effective for systemic sclerosis. METHODS: Lesional skin on the forearms of patients with systemic sclerosis (diffuse type, n =3; limited type, n =1) was exposed to medium-dose UVA1 radiation (60 J/cm(2)) daily. RESULTS: In all patients studied, UVA1 phototherapy-treated skin lesions were markedly softened after 9 to 29 exposures. Clinical improvement was associated with an increase in (1) joint passive range of motion values (P <.05), (2) skin temperature (thermography, P <.05), and (3) cutaneous elasticity (cutaneous elastometry, P <.05). Histologic evaluation of skin specimens obtained before and after UVA1 phototherapy revealed loosening of collagen bundles and the appearance of small collagen fibers. CONCLUSION: These studies indicate that UVA1 phototherapy is effective for patients with systemic sclerosis. PMID- 11004625 TI - PUVA-cream photochemotherapy for the treatment of localized scleroderma. AB - BACKGROUND: The efforts to treat localized scleroderma, including therapies with potentially hazardous side effects, are often unsatisfactory. Recently, PUVA-bath photochemotherapy has been proven highly effective in the treatment of localized scleroderma. Another form of topical PUVA therapy, 8-methoxypsoralen (8-MOP) containing cream or gel preparations, has been proven to be as effective as PUVA bath therapy for palmoplantar dermatoses. OBJECTIVE: We sought to assess the efficacy of PUVA-cream photochemotherapy in patients with localized scleroderma. METHODS: Four patients with localized scleroderma were included in the study. Diagnosis was confirmed by 20 MHz ultrasound assessment as well as pretreatment skin biopsy specimens from lesional skin. PUVA-cream therapy was performed 4 times a week; all patients received 30 treatments. RESULTS: PUVA-cream photochemotherapy induced significant clinical improvement or clearance of localized scleroderma in all patients. Clearance was documented by clinical features as well as by 20 MHz ultrasound and histopathologic analysis. CONCLUSION: PUVA-cream phototherapy can be highly effective in patients with localized scleroderma even if previous therapy was unsuccessful. PMID- 11004626 TI - Peristomal dermatoses: a novel indication for topical steroid lotions. AB - BACKGROUND: Dermatoses that interfere with the normal use of a stoma appliance are common. When preventable causes, such as infection or allergy, are not identified, barrier preparations or topical steroids have been used. However, topical medicaments formulated in a cream or ointment base will cause stoma bags to detach, resulting in leaks. OBJECTIVE: Our purpose was to investigate the efficacy and suitability of corticosteroids in aqueous/alcohol lotions in the management of peristomal dermatoses. METHODS: A clinic run by a dermatologist and 2 stoma nurses was created. Patients with a variety of noninfective, inflammatory dermatoses were treated with topical corticosteroid lotions up to a maximum of 4 weeks, with occasional use thereafter in some cases. RESULTS: Topical, aqueous/alcohol, corticosteroid lotions have been used in 60 patients and have proved particularly useful for the treatment of irritant dermatitis, pyoderma gangrenosum, psoriasis, and constitutional eczema. After the initial treatment course, occasional applications, approximately every 2 weeks, may be necessary to control the skin disorder. This low frequency of application minimizes the risk of side effects so that we have not identified local or systemic side effects in any of the patients treated so far. CONCLUSION: Topical corticosteroids formulated in aqueous alcohol lotion are effective and acceptable treatments for peristomal dermatoses. If used appropriately, the risk of side effects is low. PMID- 11004628 TI - Silicone auricular prosthesis. AB - Skin cancer surgery of the ear may result in a large defect of skin and cartilage requiring a staged flap procedure for reconstruction. An alternative to surgical reconstruction is the creation of a silicone auricular prosthesis. These prostheses provide a cost-effective and cosmetically acceptable means of camouflage for patients who decline or postpone surgical reconstruction. The process of creating a silicone auricular prosthesis is discussed. PMID- 11004627 TI - Therapeutic update: use of risperidone for the treatment of monosymptomatic hypochondriacal psychosis. AB - Monosymptomatic hypochondriacal psychoses such as delusions of parasitosis are often difficult to treat. For the past two decades, pimozide has been considered the drug of choice. Although a few controlled studies have been done to support the efficacy of pimozide, the recommendation of treatment of choice is largely based on case reports. Pimozide has significant side effects, including cardiac and extrapyramidal abnormalities, which are of most concern in the treatment of the elderly. Although pimozide does appear to be effective in some cases of somatic delusional disorders, newer antipsychotics such as risperidone have also been shown to be beneficial in treating this perplexing cluster of disorders. Risperidone is generally considered safer than pimozide and should be considered as first-line therapy for monosymptomatic hypochondriacal psychoses. PMID- 11004629 TI - Potassium iodide in dermatology: a 19th century drug for the 21st century-uses, pharmacology, adverse effects, and contraindications. AB - Potassium iodide (KI) is a useful drug in the dermatologic armamentarium. It is successfully used for inflammatory dermatoses, most notably erythema nodosum, subacute nodular migratory panniculitis, nodular vasculitis, erythema multiforme, and Sweet's syndrome. KI is also successfully used for cutaneous and lymphocutaneous sporotrichosis. The precise mechanism by which KI acts is unknown. Although many minor side effects are common with this drug, major side effects can occur in pregnant patients and those with a history of kidney or thyroid disease. This article reviews the pharmacology, mechanism of action, indications, contraindications, and adverse effects of KI as a therapeutic agent. PMID- 11004630 TI - Widespread cutaneous vascular papules associated with peripheral blood eosinophilia and prominent inguinal lymphadenopathy. AB - A new section entitled "Dermatology Grand Rounds at the NIH" appears this month. The idea behind it is to provide a unique look at instructive dermatologic cases being presented at the weekly Dermatology Branch Grand Rounds at the National Cancer Institute, National Institutes of Health (NIH), Bethesda, Maryland. Patients seen at the NIH Clinical Center (which includes the NIH inpatient wards and outpatient clinics) are indeed unusual. To be evaluated here, they must be referred by an outside physician and must be placed in a research protocol. In other words, by this criterion, all NIH patients are considered research subjects. The diseases studied at the NIH range from the common to the obscure. Similarly, the dermatologic manifestations of these diseases are equally as diverse. Thus the purpose of this new section is to share some of these fascinating cases with the dermatologic community at large, as well as to provide a glimpse of exciting translational research that has relevance to clinical dermatologists. The format will be simple. Cases will be presented as unknowns, including relevant results from the history, physical examination, and histopathologic analyses. This will be done to challenge the clinicopathologic skills of the readers; authors will make every attempt to make the color clinical and pathologic photographs of the highest quality possible to assist readers in making the correct diagnoses. The discussion will be brief, focusing on the key teaching points of each case. These teaching points will vary from cases to case, covering clinical and histologic information, treatment issues, and/or novel laboratory advances with clinical relevance. Authors will emphasize brevity and clarity in all subsections of the case report, which I hope will promote a good learning experience for dermatology residents and busy practitioners alike. This section will initially be published 4 times annually. Finally, because the section is new to the JAAD, I welcome comments and suggestions to improve the quality of the articles.-Andrew Blauvelt, MD Feature Editor PMID- 11004631 TI - Pilomatricomas contain activating mutations in beta-catenin. PMID- 11004632 TI - Directional migration of human keratinocytes in direct current electric fields requires growth factors and calcium and is regulated by epidermal growth factor receptor. PMID- 11004633 TI - Endothelin-1 down-regulates E-cadherin in melanocytic cells by apoptosis independent activation of caspase-8. PMID- 11004634 TI - The histopathologic misdiagnosis of melanoma: sources and consequences of "false positives" and "false negatives". PMID- 11004635 TI - Identifying the missing link between UV and p53: the role of phosphatidyl inositol kinase-related kinases. PMID- 11004636 TI - The use of interferon alfa as adjuvant therapy for advanced cutaneous melanoma: the need for more evidence. AB - Interferon alfa is rapidly gaining acceptance as the standard of care for patients with advanced but nonmetastatic cutaneous malignant melanoma. The randomized trials of interferons for melanoma are reviewed with attention to any survival benefits demonstrated by these studies. Because none of these studies are placebo controlled, questions regarding the placebo effects interferons may possess are addressed, as is an analogous clinical scenario in which interferons appeared to be beneficial in nonplacebo controlled trials but were shown to be ineffective in placebo-controlled trials. Moreover, given the significant toxicities and financial costs of interferons, the argument is advanced that interferon alfa should not become the standard of care for melanomas until the results of randomized, placebo-controlled trials evaluating the survival advantages of interferon alfa for melanoma become available. PMID- 11004637 TI - Surgical pearl: digital imaging for mapping Mohs surgical specimens. PMID- 11004638 TI - Paraneoplastic pemphigus with circulating antibodies directed exclusively against the pemphigus vulgaris antigen desmoglein 3. AB - A patient with follicular B-cell lymphoma presented with erythroderma associated with cutaneous and mucosal blisters. Histologic and direct immunofluorescence analysis of lesional skin showed a typical pattern of paraneoplastic pemphigus (PNP). Interestingly, indirect immunofluorescence on rat bladder was negative and immunoblot analysis of the patient's serum on epidermal extracts demonstrated antiepidermal antibodies that only recognized the pemphigus vulgaris antigen desmoglein 3, with no antibodies directed against the different proteins of the plakin family. To our knowledge this has never been reported in the literature. It exemplifies the overlap between pemphigus vulgaris and PNP and the pathogenic role of anti-desmoglein 3 antibodies in PNP. Moreover, it underscores the need to consider clinical, histologic, and immunologic features for the diagnosis of PNP. PMID- 11004639 TI - Constipation presenting as recurrent vulvovaginitis in prepubertal children. AB - Vulvovaginitis is the most common gynecologic problem in premenarcheal girls. We describe 3 patients with recurrent vulvovaginitis caused by bacterial infection who responded to treatment of constipation with resolution of the vulvovaginitis. There were no recurrences during the follow-up period of 15 to 36 months. We propose that underlying constipation should be added to the list of possible causes of recurrent vulvovaginitis. PMID- 11004640 TI - Disseminated superficial actinic porokeratosis associated with topical PUVA. AB - Disseminated superficial actinic porokeratosis (DSAP) is associated with sun exposure and tanning bed usage and has been rarely reported in association with systemic PUVA. We report the first case of DSAP occurring after topical foot PUVA. PMID- 11004641 TI - Dermoscopic features of cutaneous local recurrent melanoma. AB - We describe for the first time the dermoscopic features of cutaneous recurrent melanoma including a globular pattern and a diffuse nonhomogeneous pigmentation. These findings greatly differ from those observed in primary cutaneous melanoma and should be considered in the dermoscopic differential diagnosis of pigmented skin lesions. PMID- 11004642 TI - Photochemotherapy for systemic and localized scleroderma. PMID- 11004643 TI - Evidence-based medicine: the epistemology that isn't. PMID- 11004644 TI - Paradoxical effect of interferon alfa on lichen myxedematosus. PMID- 11004645 TI - Pruritic urticarial papules and plaques of pregnancy and multiple pregnancies. PMID- 11004646 TI - Long latency of human herpesvirus type 8 infection and the appearance of classic Kaposi's sarcoma. PMID- 11004648 TI - Neurophysiologic response to intraoperative lumbosacral spinal manipulation. AB - BACKGROUND: Although the mechanisms of spinal manipulation are poorly understood, the clinical effects are thought to be related to mechanical, neurophysiologic, and reflexogenic processes. Animal studies have identified mechanosensitive afferents in animals, and clinical studies in human beings have measured neuromuscular responses to spinal manipulation. Few, if any, studies have identified the basic neurophysiologic mechanisms of spinal manipulation in human beings or animals. OBJECTIVES: The purpose of this clinical investigation was to determine the feasibility of obtaining intraoperative neurophysiologic recordings and to quantify mixed-nerve root action potentials in response to lumbosacral spinal manipulation in a human subject undergoing lumbar spinal surgery. METHODS: An L4-L5 laminectomy was performed in a 62-year-old man. Short-duration (<0.1 ms) mechanical force, manually assisted spinal manipulative thrusts (150 N) were delivered to the lumbosacral spine with an Activator II Adjusting Instrument. With the spine exposed, spinal manipulative thrusts were delivered internally to the L5 mammillary process, L5-S1 joint, and the sacral base with various force vectors. This protocol was repeated by contacting the skin overlying respective anatomic landmarks. Mixed-nerve root recordings were obtained from gas-sterilized platinum bipolar hooked electrodes attached to the S1 nerve root at the level of the dorsal root ganglion during the spinal manipulative thrusts and during a 30 second baseline period during which no spinal manipulative thrusts were applied. RESULTS: During the active trials, mixed-nerve root action potentials were observed in response to both internal and external spinal manipulative thrusts. Differences in the amplitude and discharge frequency were noted in response to varying segmental contact points and force vectors, and similarities were noted for internally and externally applied spinal manipulative thrusts. Amplitudes of mixed-nerve root action potentials ranged from 200 to 2600 mV for internal thrusts and 800 to 3500 mV for external thrusts. CONCLUSIONS: Monitoring mixed nerve root discharges in response to spinal manipulative thrusts in vivo in human subjects undergoing lumbar surgery is feasible. Neurophysiologic responses appeared sensitive to the contact point and applied force vector of the spinal manipulative thrust. Further study of the neurophysiologic mechanisms of spinal manipulation in humans and animals is needed to more precisely identify the mechanisms and neural pathways involved. PMID- 11004649 TI - Do cerebral potentials to magnetic stimulation of paraspinal muscles reflect changes in palpable muscle spasm, low back pain, and activity scores? AB - OBJECTIVE: Previous studies have shown that cortical-evoked potentials on magnetic stimulation of muscles are influenced by muscle contraction, vibration, and muscle spasm. This study was carried out to determine whether these potentials correlate with palpatory muscle spasm, patient symptoms, and disability in patients with low back pain. METHODS: A prospective observational study was performed on 13 subjects with a history of low back pain visiting an orthopedic hospital-based clinic. Patients were screened for serious pathologic conditions by an orthopedic surgeon. The patients were then evaluated for the presence of muscle spasm by one of the investigators who was blinded to the results of the evoked potential studies. Patients were asked to complete a low back pain visual analogue scale (VAS) and a Roland-Morris Activity Scale (RMAS). Cortical-evoked potentials were recorded with a magnetic stimulator placed over the lumbar paraspinal muscles with the patient in the prone position. The palpatory examination, VAS, RMAS, and the cortical potentials were repeated after 2 weeks of therapy commonly used to reduce muscle spasm. RESULTS: The patients demonstrated a significant decrease in low back pain VAS and RMAS scores after treatment compared with before treatment. There was a reduction in the amount of palpatory muscle spasm in 11 of 13 cases. The cortical potentials before treatment were attenuated compared with previously reported controls and showed a significant increase before and after treatment in the amplitude of these potentials with multivariate analysis of variance. There was significant correlation between the changes in cortical potentials after treatment and the changes noted in paraspinal muscle spasm and VAS and RMAS scores. CONCLUSIONS: This study confirms the previous report that the amplitude of cerebral-evoked potentials on magnetic stimulation of paraspinal muscles is depressed in the presence of palpable muscle spasm. The close correlation among these potentials, paraspinal muscle spasm, and clinical symptoms suggests that the measurement of muscle activity may be more important in the assessment of low back pain than is commonly accepted. PMID- 11004650 TI - Short-term effects of cervical manipulation on edge light pupil cycle time: a pilot study. AB - BACKGROUND: Edge light pupil cycle time (ELPCT) is one of the eye's light reflexes. Studies have shown ELPCT to be a measurable constant, unaffected by visual acuity, refractive error, eye color, pupil size, or sex. Control of this reflex occurs through the autonomic nervous system. Various authors suggest that spinal manipulative techniques can produce distant effects mediated in part by alterations in autonomic tone after intervention. OBJECTIVE: To investigate the effects of a C1-2 high-velocity, low-amplitude manipulation on ELPCT. DESIGN: A single-group, randomized pilot study without a control group. METHODS: Thirteen men (mean age 24.2 years) without a history of eye disease or central or autonomic nervous system pathologic conditions had their ELPCT measured before and after manipulation. The manipulation comprised a high-velocity, low-amplitude rotatory thrust, localized to the C1-2 joint on the left (n = 6) or right (n = 7) eye, determined randomly. RESULTS: ELPCT measures demonstrated a significant difference for both eyes before and after manipulation (P = .002; the right eye, P = .027; the left eye, P = .046). CONCLUSION: This suggests that ELPCT, which is mediated through the autonomic nervous system, can be directly influenced by high velocity manipulation to the upper cervical spine. PMID- 11004651 TI - MRI of the spine and spinal cord: imaging techniques, normal anatomy, artifacts, and pitfalls. AB - BACKGROUND: Magnetic resonance imaging (MRI) is widely used to evaluate the spine and spinal cord. OBJECTIVE: In this article, MRI of the spine is discussed in terms of normal anatomy, standard and advanced imaging techniques, general indications, limitations, and potential for the future. DISCUSSION: Although MRI does not provide the high bony detail possible with computed tomography, the appropriate combination of the sequences takes advantage of the different tissue characteristics to discriminate the various bony-and soft-tissue structures of the spine. CONCLUSION: MRI enables the imaging specialist to evaluate a large anatomic region in multiple planes and can better examine the spinal cord. PMID- 11004652 TI - Chiropractic biophysics digitized radiographic mensuration analysis of the anteroposterior cervicothoracic view: a reliability study. AB - OBJECTIVE: To investigate the reliability of a radiographic measurement procedure that uses a computer and sonic digitizer to determine projected spinal displacements from an ideal, normal position. DESIGN: A blind, repeated-measure design was used. Anteroposterior cervicothoracic spine radiographs were presented in random order to each of 3 examiners. Each film was digitized, and the films were randomized for a second examination. SETTING: Private, primary care chiropractic clinic. MAIN OUTCOME MEASURES: Intraclass correlation coefficients for intraexaminer and interexaminer reliability for measures on radiographs comparing the perpendicular distance (T(x)) from a vertical axis line drawn through the center of T4 and the center of C2, the linear distance (vertebra(apex)) from the center of the vertebra most displaced from a line connecting the centers of C2 and T4, the angle (Rz) formed by the intersection of the vertical axis line and the upper thoracic line, and the angle of intersection (CDA) between the upper thoracic line and the cervical line. RESULTS: Intraexaminer reliability for T(x) distance was 0.99 to 1.00, with confidence intervals from 0.98-1.00; for vertebra(apex) was 0.96 to 0.97, with confidence intervals from 0.92-0.98; for Rz was 0.94 to 0.98, with confidence intervals from 0. 89-0.99; and for CDA was 0.92 to 0.95, with confidence intervals from 0.84 0.97. Interexaminer reliabilities for the 3 examiners ranged from 0.97 to 0.99. CONCLUSIONS: Measures similar to those described in this study are commonly used to quantify and categorize spinal displacements from true vertical alignment (i.e., scoliosis measurements). Intraclass correlation coefficient values >0.70 are considered accurate enough for use in clinical and research applications. The measures tested here would fit within these guidelines of reliability. Establishing reliability is an important first step in evaluating these measures so that future studies of validity may be undertaken. PMID- 11004653 TI - Lumbosacral transitional segments: classification, prevalence, and effect on disk height. AB - OBJECTIVE: To determine the rate of lumbosacral transitional segments among chiropractic practice settings and to determine if this anomaly would affect the height of the lumbosacral disk. STUDY DESIGN: Retrospective review of radiographs. SETTING: Los Angeles College of Chiropractic outpatient clinic and a private chiropractic office. SAMPLES: A total of 20 lumbar series with lumbosacral transitional segments from a private chiropractic office, 47 lumbar series with lumbosacral transitional segments, and 60 age- and sex-matched control series from a college clinic. RESULTS: A total of 2.3% of 882 lumbar series at the private chiropractic office and 6.5% of 786 lumbar series at the Whittier Health Center showed lumbosacral transitional segments of types II, III, or IV. The L5-S1 intervertebral disk height was significantly smaller in patients with lumbosacral transitional segments compared with those without (ie, 11% vs 19% of total lumbar disk height, respectively). When bilateral bony fusion of L5 to the sacrum was present, the L5-S1 disk height was significantly smaller than that without bony fusion (ie, 8% vs. 12% to 14% of total lumbar disk height, respectively). CONCLUSIONS: We conclude that the rate of occurrence of lumbosacral transitional segments is low in chiropractic practice. In the presence of lumbosacral transitional segments, especially when there was bony fusion, the lumbosacral intervertebral disk was significantly narrower than the upper lumbar disks, which should not be considered as disk degeneration or displacement. The type of lumbosacral transitional segment present also showed a significant effect on the height of the lumbosacral disk. PMID- 11004654 TI - Complex regional pain syndrome and chiropractic. AB - OBJECTIVE: Complex regional pain syndromes (CRPS) represent curious and difficult syndromes for both patient and clinician. CRPS presents as a triad of signs and symptoms, usually after a seemingly trivial injury to a peripheral joint or appendage. The clinical triad includes severe pain, vasomotor changes in and around the affected area, and trophic changes in the affected limb. Many of the acute symptoms are similar to those seen after many acute injuries, which makes an early diagnosis often times difficult. Current treatment protocols revolve around aggressive physical therapy plus pharmacologic interventions aimed at limiting sympathetic nervous system activity. OBJECTIVE: To review the literature on CRPS regarding symptoms, diagnosis, treatment, and causal mechanisms and to discuss alternative treatment approaches and the possible role of chiropractic care in patient rehabilitation. DATA SOURCES: Texts, review articles, and randomized clinical trials investigating treatments, causes, and epidemiology. CONCLUSIONS: Recent research calls into question the predominant theories that view excessive sympathetic nervous system activity as the cause of CRPS. No evidence of an increase in sympathetic nervous system activity has been found, and new theories suggest that an increase in the sensitivity of neurotransmitter receptors may be the cause of CRPS. Alternatively, other research has suggested that a local inflammatory process may in fact cause CRPS. Although no research has been completed examining the role of chiropractic care in the treatment of CRPS, there is reason to believe that spinal manipulation may be beneficial to patients with CRPS. PMID- 11004655 TI - Clinical considerations in the chiropractic management of the patient with Marfan syndrome. AB - OBJECTIVE: To describe the chiropractic management of a patient with whiplash associated disorder and a covert, concomitant dissecting aneurysm of the thoracic aorta caused by Marfan syndrome or a related variant. CLINICAL FEATURES: A 25 year-old man was referred by his family physician for chiropractic assessment and treatment of neck injuries received in a motor vehicle accident. After history, physical examination, and plain film radiographic investigation, a diagnosis of whiplash-associated disorder grade I was generated. INTERVENTION AND OUTCOME: The whiplash-associated disorder grade I was treated conservatively. Therapeutic management involved soft-tissue therapy to the suspensory and paraspinal musculature of the upper back and neck. Rotary, manual-style manipulative therapy of the cervical and compressive manipulative therapy of the thoracic spinal column were implemented to maintain range of motion and decrease pain. The patient achieved full recovery within a 3-week treatment period and was discharged from care. One week after discharge, he underwent a routine evaluation by his family physician, where an aortic murmur was identified. Diagnostic ultrasound revealed a dissecting aneurysm measuring 78 mm at the aortic root. Immediate surgical correction was initiated with a polyethylene terephthalate fiber graft. The pathologic report indicated that aortic features were consistent with an old (healed) aortic dissection. There was no evidence of acute dissection. Six month follow-up revealed that surgical repair was successful in arresting further aortic dissection. CONCLUSION: The patient had an old aortic dissection that pre-dated the chiropractic treatment (which included manipulative therapy) for the whiplash-associated disorder. Manipulative therapy, long considered an absolute contraindication for abdominal and aortic aneurysms, did not provoke the progression of the aortic dissection or other negative sequelae. The cause, histology, clinical features, and management considerations in the treatment of this patient's condition(s) are discussed. PMID- 11004656 TI - Residency programs at the Canadian Memorial Chiropractic College. PMID- 11004657 TI - Reflex effects of vertebral subluxations: the peripheral nervous system. An update. PMID- 11004658 TI - Reflex effects of vertebral subluxations: the peripheral nervous system. An update PMID- 11004659 TI - Motion palpation: it's time to accept the evidence. PMID- 11004660 TI - Motion palpation: It's time to accept the evidence PMID- 11004661 TI - A new era of cancer research: cutting and pasting of chromosomes. PMID- 11004662 TI - K-ras point mutation is associated with enhancement by deoxycholic acid of colon carcinogenesis induced by azoxymethane, but not with its attenuation by all-trans retinoic acid. AB - The effects of deoxycholic acid (DCA) with and without all-trans-retinoic acid (ATRA) on the incidence of colon tumors induced by azoxymethane, the incidence of K-ras point mutation in colon tumors and the labeling index of colon mucosa were investigated in male Wistar rats. Rats received 5 weekly injections of 7.4 mg/kg body weight of azoxymethane. From the start of the experiment, all rats in 3 groups also received chow pellets containing 0.3% DCA with and without s.c. injections of 0.75 or 1.5 mg/kg body weight of ATRA every other day until the end of week 45. Oral administration of DCA significantly increased the incidence of colon tumors in week 45. Concomitant use of DCA and ATRA at either dose significantly attenuated the enhancement by DCA of colon tumorigenesis. Administration of DCA significantly increased the incidence of K-ras point mutation in colon tumors and the labeling index in the colon mucosa. Combined administration of DCA and ATRA significantly reduced the labeling index of colon mucosa, which was increased by DCA, but did not affect the incidence of K-ras point mutation in colon tumors. These findings suggest that DCA enhances development of colon tumors and that this enhancement is attenuated by ATRA. A possible mechanism of this enhancement is induction of K-ras point mutation. However, decreased cell proliferation in the colon mucosa may be closely related to the attenuation of DCA-enhanced colon tumorigenesis, but not suppression of K ras point mutation. PMID- 11004663 TI - Tumor-derived p53 mutant C174Y is a gain-of-function mutant which activates the fos promoter and enhances colony formation. AB - SV40 large T antigen-induced primitive neuroectodermal tumors of the rat provide a model system to study induction and progression of primitive neuroectodermal tumors at the molecular level. A cell line derived from such a tumor reproducibly gave rise to malignant derivatives that ceased large T-antigen expression but harbored a mutant p53 allele with a common mutation at Cys(174) to Tyr (C174Y). To determine whether this p53 mutation contributes to tumor progression, we analyzed mutant C174Y functionally. Co-transfection experiments in Saos-2 cells with mutant or wild-type p53 and reporter genes linked to various p53-responsive promoters revealed that mutant C174Y failed to transcriptionally transactivate the Mdm2, Waf1, Cyclin G and Bax promoters. Loss of transcriptional activation correlated with loss of DNA-binding activity. Moreover, mutant C174Y exhibited a dominant negative effect on co-expressed wild-type p53. The ability of mutant p53 to repress the viral RSV, LTR or SV40 early promoters or the cellular fos promoter was likewise impaired. In contrast, it showed even induction of the fos promoter. Consistent with these observations, mutant C174Y was non-functional in the suppression of Saos-2 cell growth and even conferred a growth advantage to the cells. Surprisingly, mutant C174Y was also impaired in nuclear transport, as revealed by immunofluorescence analyses. Taken together, our results demonstrate that mutant C174Y possesses features that can positively contribute to cancer progression. PMID- 11004664 TI - Gene expression of neuronal nitric oxide synthase and adrenomedullin in human neuroblastoma using real-time PCR. AB - The objective of our study was to assess the gene expression of the antiproliferative systems neuronal nitric oxide synthase (nNOS) and adrenomedullin (AM) in human neuroblastoma. A novel real-time PCR method was evaluated using neuropeptide Y (NPY) for validation. Glyceraldehyd-3-phospate dehydrogenase (GAPDH) and NPY gene expression in neuroblastomas of 50 patients were measured in parallel by competitive quantitative and TaqMan real-time RT PCR. AM and nNOS mRNA were determined by real-time PCR. Our results showed a linear relationship between competitive quantitative and real-time RT-PCR measurements of NPY and GAPDH (r = 0.87 and r = 0.92, respectively). AM and nNOS mRNA was found in all tumor samples. AM/GAPDH mRNA increased with higher differentiation according to Shimada (p = 0.013). There was no relation between MYCN amplification nor with the tumor stage (p = 0.78 and p = 0.30, respectively). AM/GAPDH did not relate to recurrence or death in a 5-year follow up period. Neuronal NOS/GAPDH expression did not relate to any biological or clinical parameter of prognosis or differentiation. Similar results were obtained when the neuronal marker protein gene product 9.5 (PGP9.5) was used to normalize mRNA concentration. In conclusion, TaqMan real-time PCR appears to be a reliable method to quantify gene expression in neuroblastomas. Adrenomedullin mRNA in neuroblastoma is linked to tumor differentiation but not to prognostic markers. PMID- 11004665 TI - Selective activation of the fatty acid synthesis pathway in human prostate cancer. AB - A substantial subset of breast, colorectal, ovarian, endometrial and prostatic cancers displays markedly elevated expression of immunohistochemically detectable fatty acid synthase, a feature that has been associated with poor prognosis and that may be exploited in anti-neoplastic therapy. Here, using an RNA array hybridisation technique complemented by in situ hybridisation, we report that in prostate cancer fatty acid synthase expression is up-regulated at the mRNA level together with other enzymes of the same metabolic pathway. Contrary to the observations that in many cell systems (including androgen-stimulated LNCaP prostate cancer cells) fatty acid and cholesterol metabolism are co-ordinately regulated so as to supply balanced amounts of lipids for membrane biosynthesis, storage or secretion, no changes in the expression of genes involved in cholesterol synthesis were found. These findings point to selective activation of the fatty acid synthesis pathway and suggest a shift in the balance of lipogenic gene expression in a subgroup of prostate cancers. PMID- 11004666 TI - Alterations in the gene expression profile of MCF-7 breast tumor cells in response to c-Jun. AB - MCF7 breast tumor cells overexpressing human c-Jun exhibit a transformed phenotype characterized not only by increased tumorigenicity but also by enhanced motility and invasion. The cellular phenotypic response to c-Jun overexpression is likely due, at least in part, to altered patterns of gene expression. In order to begin to understand the complexities by which elevated production of c-Jun alters the state of the cell, we have profiled the expression of 588 different genes by comparative hybridization. By using this approach, we have identified a total of 21 upregulated or downregulated gene targets responsive to c-Jun overexpression. Interestingly, 8 of these genes have been previously found associated with c-Jun or AP-1 activity and therefore provide internal validation for this approach to target gene discovery. The remaining 13 genes represent potential new c-Jun regulated target genes. Genomic sequence information was available for 15 of the 21 genes identified in this screen. Analysis of these genomic sequences revealed the presence of AP-1 or AP-1-like sequences in 12 of the 15 genes examined. Consistent with a direct mechanism of target regulation by c-Jun, gel shift analysis of selected AP-1-containing promoter regions revealed elevated and specific binding by proteins present in nuclear extracts of c-Jun expressing MCF7 cells. PMID- 11004667 TI - p53-dependent induction of heat shock protein 27 (HSP27) expression. AB - Transcriptional activation of the p53 target genes plays a critical role in the cellular response to DNA damage, hypoxia, cellular stress and other signals regulating the cell cycle and apoptosis. The discovery of new p53 target genes continues to reveal novel mechanisms of action of this multifaceted protein. We used cDNA arrays to search for p53-regulated genes in prostate cancer cells. In this report, we describe robust induction of heat shock protein 27 (hsp27) in prostate cancer cells (DU145, LNCaP, PC3) following wild-type p53 expression from an adenoviral p53 expression vector (AdWTp53). A mutant p53 (R175H)-containing adenoviral expression vector did not induce hsp27. hsp27 expression was not altered in prostate cancer cells following expression of cyclin-dependent kinase inhibitors: p21(waf1/cip1) and p27(kip1) from adenoviral expression vectors. Treatment of cells with staurosporine, an apoptosis-inducing agent, did no affect hsp27 expression. These observations provide evidence that induction of hsp27 expression was wild-type p53-specific and was not due to non-specific effects of cell growth arrest and/or apoptosis. Previous studies and the experiment reported here show induction of hsp27 expression in response to androgen ablation, a physiological state that induces apoptosis in prostatic epithelial cells. The nature of p53 and hsp27 interactions in the regulation of apoptosis and/or cell growth needs to be further defined. PMID- 11004668 TI - Protein kinase C isoforms involved in regulation of cell shape and locomotion of human fibrosarcoma HT1080 cells. AB - The role of protein kinase C (PKC) isoforms in the regulation of cell shape [switch between fibroblast-like and crescent shape (CS)] and of locomotion of human fibrosarcoma HT1080 cells has been investigated. The PKC activator phorbol myristate acetate (PMA) induced the transition of elongated fibroblast-like cells into CS cells and stimulated locomotion. Both responses to PMA were inhibited by the PKC inhibitor Ro 31-8220. Analysis of the time course showed that stimulation of shape changes (formation of CS cells) and locomotor activity (increase in the proportion and speed of locomoting cells) was maximal in the early phase of the response (up to 2.5 hr) and significantly decreased later (15 to 21 hr). CS formation and stimulated locomotion correlated closely with a marked redistribution from the cytosol to the membrane of PKC isoforms alpha, beta1 and gamma in the early phase (0.5 to 2 hr) following activation with PMA. The subsequent reduction of the proportion of CS cells and of cell locomotion correlated with down-regulation of these isoforms in the second phase (16 to 21 hr). In contrast, the total amount and distribution of PKC beta2 remained almost unchanged with 10(-8) M PMA up to 21 hr. Furthermore, changes in shape and locomotion did not correlate with the responses of PKC delta to PMA. Inhibition of PMA-stimulated locomotion by the more specific inhibitor Go 6976 is consistent with a role of PKC alpha and beta1 in this response. Ro 31-8220 alone induced a moderate down-regulation of PKC isoforms alpha and delta, but it also inhibited the more pronounced down-regulation of these isoforms by PMA. Our results indicate that activation of PKC isoforms alpha, gamma and beta1, but not beta2 or delta, stimulates locomotion and formation of CS cells in human fibrosarcoma HT1080 cells. PMID- 11004669 TI - Association between loss of heterozygosity of BRCA1 and BRCA2 and morphological attributes of sporadic breast cancer. AB - Germline mutations in the breast cancer-associated genes BRCA1 and BRCA2 confer a lifetime risk of malignancy. Distinctive morphological features have been attributed to these familial tumours; however, in sporadic breast cancer, the inter-relationship between loss of heterozygosity (LOH) of these loci and tumour morphology remains to be fully elucidated. We studied a series of 120 sporadic breast carcinomas using microsatellite markers to identify LOH of BRCA1, BRCA2, p53 and PTEN. The associations between loss at each of the loci were examined and related to tumour morphology. LOH of the 4 loci did not occur independently; there were highly significant associations between LOH of BRCA1 and both BRCA2 (p < 0.001) and p53 (p < 0.001). LOH at all 4 loci was significantly associated with a high degree of nuclear pleomorphism. Tumours with LOH of BRCA1 also had high mitotic indices, few tubules and a paucity of DCIS, all of which are morphological features similar to those described for familial cases. Following Bonferroni's correction for multiple tests, we found that the tumours with LOH of BRCA1 were still significantly associated with a high mitotic index (p = 0.0006) and a high degree of nuclear pleomorphism (p = 0.001). PMID- 11004670 TI - Variant estrogen receptor alpha mRNAs in human breast cancer specimens. AB - A panel of human breast cancer specimens was examined for single base change mutations by DNA sequencing and for larger deletions using a PCR-based assay. In the cancer specimens examined, no sequencing variants were detected other than a previously characterized polymorphism. Although most of the specimens contained estrogen receptor (ER) variants at a low level, 2 of 118 specimens exhibited variants which, after amplification, constituted most of the amplified ER cDNA. One specimen contained a single variant form, and there was little evidence of the wild-type ER mRNA by PCR, Northern blotting or immunocytochemistry. The second specimen, despite the presence of a normal-sized mRNA by Northern blotting and normal immunocytochemical staining for ER, contained at least 5 different variant forms as well as the wild-type ER. All but 1 of the variant forms were processing variants, and 3 of these processing variants have not been described before. One variant, although lacking exons 2-4, has break points in exons 1 and 5 that do not correspond to intron-exon boundaries. This variant might reflect more widespread damage to the genome in this breast cancer specimen. The low level of occurrence of variants suggests that ER variant forms, at least in the coding region, do not contribute generally to the progression of breast cancer. PMID- 11004671 TI - Primary cervical carcinomas show 2 common regions of deletion at 3P, 1 within the FHIT gene: evaluation of allelic imbalance at FHIT, RB1 and TP53 in relation to survival. AB - Chromosome arm 3p is re-arranged in many tumor types, including cervical carcinomas. Putative tumor-suppressor genes on 3p have been proposed, including the FHIT gene, which maps to chromosome band 3p14.2. We have analyzed 79 primary cervical carcinomas for allelic imbalance (AI) at 17 chromosome 3 loci, including 3 within the FHIT gene. Expression of the FHIT gene was evaluated after immunohistochemistry with an antibody against the pFHIT protein. Previously determined human papillomavirus status, defined after in situ hybridization, showed type 16 or 18 in 56/77 tumors. Tumors were also analyzed for AI at loci within the RB1 (chromosome band 13q14.2) and the TP53 (17p13) genes for AI. AI was found at 1 or more 3p loci in 50/79 tumors, at frequencies ranging from 30% to 52% at the individual loci. Two smallest regions of overlapping deletion (SROs) were found, 1 including parts of the FHIT gene (SRO flanked by D3S1481 and D3S1313) and another more distal SRO between D3S32 and D3S1286. FHIT protein expression was reduced in 57/69 (83%) tumors but not associated with AI at FHIT loci (p = 0.56). AI was found in TP53 and RB1 in 18% and 29% of the samples, respectively. Relapse-free survival was associated with AI in the TP53 gene in both a univariate (p = 0.0003) and a multivariate (p = 0.004) analysis. This study confirms a high frequency of AI at chromosome arm 3p in primary cervical carcinomas. The AI results and the reduced FHIT protein staining indicate that FHIT alterations are important in cervical carcinogenesis. PMID- 11004672 TI - TP53 mutations and MDM2 gene amplification in squamous-cell carcinomas of the esophagus in south Thailand. AB - Squamous-cell carcinoma of the esophagus (SCCE) shows geographic variations in incidence that are thought to reflect the etiological involvement of environmental or dietary risk factors. Mutations of TP53 are frequent in SCCE, and there is evidence that both the frequency and type of these mutations may differ from one geographic area to the other. Although SCCE is relatively rare in most parts of Thailand, the province of Songkhla (south Thailand) has been described as a high-risk area for SCCE. We have analyzed 56 SCCE cases from this area for TP53 mutations by denaturing gradient gel electrophoresis (DGGE, exons 5 8) and direct DNA sequencing. The same tumors were also analyzed for MDM2 gene amplification by differential PCR. TP53 mutations were detected in 23 cases (41%). In contrast, clear amplification of MDM2 was detected in only 2 cases (4%), both of which contained wild-type TP53. Comparison with published results from other geographic areas of high SCCE incidence revealed that the spectrum of TP53 mutations in south Thailand is similar to that observed in central China (Henan Province) but clearly differs from that of SCCE from western Europe (Normandy, France; northern Italy), with more G:T transversions and fewer mutations affecting A and T base pairs. These results suggest that SCCE from south Thailand and from central China may involve similar risk factors. PMID- 11004673 TI - Allelic loss in human intrahepatic cholangiocarcinoma: correlation between chromosome 8p22 and tumor progression. AB - Intrahepatic cholangiocarcinoma (ICC) is the second most common malignant primary tumor of the liver in Japan. Despite progress in operative techniques and adjuvant therapy, the prognosis of ICC remains very poor. Therefore, it is important to investigate the mechanism of carcinogenesis and progression of ICC. We screened allelic losses at 6 loci, including that of novel tumor-suppressor gene FEZ1 on chromosome 8p, and at 5 microsatellite loci to define the association with tumor-suppressor genes (HNPCC, APC, RB1, p53, DCC) in tumors from 18 unrelated ICC patients by PCR-loss of heterozygosity (LOH) assay and correlated the alterations with clinicopathological parameters. As a result, 61.1% (11 of 18) of patients showed LOH at 1 of the loci at least, and microsatellite instability was observed in 16.7% (3 of 18). At locus D8S258, relatively frequent LOH was detected (17.6%) compared with other loci on chromosome 8p. Among the other 5 chromosomal arms tested, the highest frequency of LOH (23.5%) was observed at D17S153. Fifty percent of cases with the mass forming + periductal infiltrating type were frequently detected by LOH at D8S258 compared to cases of the mass-forming or intraductal growth type. In conclusion, we show that 1 putative tumor-suppressor gene on 8p22 may relate to progression of ICC and suggest that the p53 tumor-suppressor gene may be associated with carcinogenesis of ICC. PMID- 11004674 TI - Immunization of cancer patients with autologous cancer-derived heat shock protein gp96 preparations: a pilot study. AB - Heat shock protein (HSP)-peptide complexes isolated from murine cancers elicit protective immunity and T lymphocytes specific for the cancer from which the HSPs are isolated. A pilot study was designed to test the feasibility, immunogenicity and toxicity of such treatment in cancer patients. Sixteen patients with assorted advanced malignancies, which had become refractory to established therapies, were recruited. The gp96 vaccine was prepared for each patient from tumor obtained from that patient. Anti-tumor immune responses were evaluated using Elispot assays of T cells in peripheral blood after minimal in vitro stimulation. No unacceptable vaccine-related toxicities or auto-immune reactions were observed. Immunization with autologous gp96 elicited MHC I-restricted, tumor-specific CD8(+) T lymphocytes in 6/12 patients immunized. In addition, expansion of the NK cell population was seen in 8/13 of patients immunized. These observations are entirely consistent with the murine experience and form a firm basis for future trials with clinical end points, using autologous, patient-specific HSP-peptide vaccines. PMID- 11004675 TI - Defective antigen presentation resulting from impaired expression of costimulatory molecules in breast cancer. AB - Previous experiments from our laboratory have shown that immune mechanisms aiming at the destruction of tumour cells including the recognition of target cells and their elimination via the expression of intercellular adhesion molecule-1 (ICAM 1; CD54), the production of tumour necrosis factor-alpha (TNF-alpha) by monocytes and appropriate function of lymphocyte subpopulations were defective in breast cancer. Previous observations were extended to assess expression levels and regulatory mechanisms of costimulatory molecules CD54, CD80 and CD86 on monocytes derived from patients with early breast cancer (EBC). In addition, antigen presentation by antigen-presenting cells (APC) was analyzed within this context. We report that monocytes derived from patients with EBC exhibited significantly decreased expression levels of CD54 (p = 0.0002), CD80 (p = 0.009) and CD 86 (p = 0.002) compared with monocytes derived from healthy females. Simultaneously, lipopolysaccharide (LPS)-induced TNF-alpha production of monocytes was found to be defective in patients with EBC. Finally, T-cell proliferation in response to tetanus toxoid (TT) was significantly decreased in patients with EBC compared with healthy control females (p < 0.0001). Furthermore, T-cell proliferation in response to TT-pulsed APC derived from healthy controls was significantly inhibited in the presence of anti-CD54 and/or anti-CD80 antibodies in a dose dependent manner, thus corroborating the necessity of the presence of CD54 and CD80 as costimulatory molecules in the present setting. We conclude that monocytes derived from patients with EBC showed a simultaneous defect of expression of CD54 and its regulation via TNF-alpha, CD80 and CD86 as well as T cell proliferation following exposure to TT-pulsed APC. Based upon these findings, it is speculated that defects in costimulatory molecule expression might contribute to tolerance of the immune system towards the presence of malignant cells in patients with EBC. PMID- 11004676 TI - Tributyrin induces differentiation, growth arrest and apoptosis in androgen sensitive and androgen-resistant human prostate cancer cell lines. AB - Progression to androgen independence remains the main problem that impacts on survival and quality of life in prostate cancer patients. We have investigated the potency of tributyrin, an orally available prodrug of butyrate, to induce growth arrest, differentiation and apoptosis in LNCaP, PC-3 and TSU-PR1 human prostate cancer cell lines. Cells were treated with 0.1 to 5 mM tributyrin or sodium butyrate. Growth inhibition, cell cycle arrest and apoptosis induction was assessed using standard methods. Both agents induced a more differentiated, fibroblast-like phenotype in androgen-sensitive as well as androgen-resistant cell lines. Expression of prostate-specific antigen was increased in LNCaP cells by tributyrin as a indicator of differentiation. The IC(50) for sodium butyrate was 2.5 mM in PC-3 and TSU-PR1 cells. LNCaP cells exhibited <50% growth inhibition at 5 mM sodium butyrate. However, the IC(50) for tributyrin was 0.8 mM in PC-3 cells, 1.2 mM in TSU-PR1 cells and 3.1 mM in LNCaP cells. Flow cytometry revealed a strong G1-arrest after exposure to tributyrin or sodium butyrate. Both agents resulted in a strong increase of apoptosis rates compared with mock treated cells. Overall, tributyrin had a 2.5- to 3-fold growth inhibitory and apoptosis-inducing potency compared with equimolar concentrations of sodium butyrate. Our results demonstrate that tributyrin is more potent than butyrate in regard to cell growth inhibition and apoptosis induction at pharmacologically relevant concentrations. Hence, tributyrin may be a promising candidate for clinical protocols in prostate cancer. PMID- 11004677 TI - Apoptotic responsiveness of the Ewing's sarcoma family of tumours to tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). AB - We investigated the cytotoxic responsiveness of 40 cell lines derived from representatives of the Ewing's sarcoma family of tumours (ESFT), i.e., Ewing's sarcoma (ES), peripheral primitive neuroectodermal tumour (pPNET) and Askin tumour (AT), to tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). Incubation with TRAIL at 100 ng/ml induced cell death at 24 hr in 19 of 26 ES, 11 of 12 pPNET and 2 of 2 AT cell lines. Half-maximal cell death concentrations (IC(50) values) varied from 0.1 to 20 ng/ml. TRAIL displayed potent cytotoxic activity against freshly derived ESFT cell isolates. Cytotoxicity was associated with phosphatidylserine expression and internucleosomal DNA fragmentation, features characteristic of apoptosis. The apoptotic programme in the sensitive ESFT VH-64 cell line revealed TRAIL-induced activation of FLICE/MACH1 (caspase-8) and CPP32/Yama/apopain (caspase-3) and processing of the prototype caspase substrate poly(ADP-ribose) polymerase. In addition, TRAIL provoked a collapse of the mitochondrial transmembrane potential (DeltaPsi(m)), parallelled by a reduction in ATP levels and release of cytochrome c from mitochondria into the cytosol. Inhibition of caspase-8 and caspase-3 by zIETDfmk and zDEVDfmk, respectively, substantially prevented TRAIL-induced apoptosis. However, zIETDfmk, but not zDEVDfmk, reduced TRAIL-mediated DeltaPsi(m) dissipation, indicating that TRAIL causes mitochondrial dysfunction through caspase-8 acting upstream of mitochondria. While macromolecule synthesis inhibitors (actinomycin D, cycloheximide) augmented susceptibility to TRAIL in TRAIL-responsive cell lines, these agents did not render TRAIL-resistant cell lines susceptible to TRAIL. However, the proteasome inhibitor MG132 sensitised to TRAIL in resistant cell lines. Collectively, these results show that TRAIL initiates effective death in the vast majority (80%) of cell lines derived from ESFT. Since TRAIL provoked cell death in ESFT ex vivo, this cytokine may be a promising drug for the treatment of ESFT in vivo. PMID- 11004678 TI - New highly lipophilic camptothecin BNP1350 is an effective drug in experimental human cancer. AB - BNP1350, 7-[(2-trimethylsilyl)ethyl]-20(S)-camptothecin, is a novel semi synthetic, highly lipophilic, silicon-containing camptothecin and an inhibitor of topoisomerase I. It has been supercomputer engineered for superior oral bioavailability, superior lactone stability, broad anti-tumor activity, increased potency and insensitivity to Pgp/MRP/LRP drug resistance. We determined the efficacy of BNP1350 in experimental human colon cancer and compared its anti tumor effects with those of CPT-11/SN-38. We also determined a possible influence of Pgp, MRP and LRP on the efficacy of BNP1350. The in vitro anti-proliferative capacity of the compounds using various exposure times was assessed in five colon cancer cell lines and indicated that BNP1350 was similarly effective or slightly more potent than SN-38. Four cell lines of other origin with sublines expressing Pgp, MRP and/or LRP showed that BNP1350 was significantly more effective than SN 38 (p < 0.05) and that the activity of BNP1350 was not reduced in multidrug resistant cells. For in vivo experiments, BNP1350 was given 1.0 mg/kg i.p. or 1.5 mg/kg p.o. daily x 5 and CPT-11 20 mg/kg i.p. daily x 5 being equitoxic schedules in nude mice bearing s.c. human tumor xenografts. The schedules were studied in colon cancer xenografts COLO320, COLO205 or WiDr as well as in two Pgp-positive xenografts 2780AD and BRO/mdr1.1 and the parental Pgp-negative A2780 ovarian cancer xenografts and BRO melanoma xenografts. Growth inhibition of >50% was obtained for BNP1350 given i.p. in six out of the seven xenografts studied. BNP1350 was similarly effective when given i.p. or p.o. CPT-11 was as effective as BNP1350, except in BRO and BRO/mdr1.1 xenografts. Pgp expression in xenografts in vivo confirmed that there was no negative influence on the efficacy of BNP1350. In conclusion, BNP1350 shows a broad spectrum of activity in experimental human tumors and is a suitable candidate for oral treatment of cancer. PMID- 11004679 TI - A novel recombinant fusion toxin targeting HER-2/NEU-over-expressing cells and containing human tumor necrosis factor. AB - Over-expression of the proto-oncogene HER2/neu in breast cancer and certain other tumors appears to be a central mechanism that may be partly responsible for cellular progression of the neoplastic phenotype. Transfection of mammalian cells and over-expression of HER2/neu appears to result in reduced sensitivity to the cytotoxic effects of tumor necrosis factor (TNF) and reduced sensitivity to immune effector killing. The single-chain recombinant antibody sFv23 recognizes the cell-surface domain of HER2/neu. The cDNA for this antibody was fused to the cDNA encoding human TNF, and this sFv23/TNF fusion construct was cloned into a plasmid for expression in Escherichia coli. The fusion protein was expressed and purified by ion-exchange chromatography. SDS-PAGE demonstrated a single band at the expected m.w. (43 kDa). Western analysis confirmed the presence of both the antibody component and the TNF component in the final fusion product. The fusion construct was tested for TNF activity against L-929 cells and found to have biological activity similar to that of authentic TNF (SA 420 nM). The scFv23/TNF construct bound to SKBR-3 (HER2-positive) but not to A-375 human melanoma (HER2 negative) cells. Cytotoxicity studies against log-phase human breast carcinoma cells (SKBR-3-HP) over-expressing HER2/neu demonstrate that the sFv23/TNF fusion construct was 1, 000-fold more active than free TNF. Tumor cells expressing higher levels of HER2/neu (SKBR-3-LP) were relatively resistant to both the fusion construct and native TNF. These studies suggest that fusion constructs targeting the HER2/neu surface domain and containing TNF are more effective cytotoxic agents in vitro than native TNF and may be effective against tumor cells expressing intermediate, but not high, levels of HER2/neu. PMID- 11004680 TI - Cancer cell-targeted drug delivery utilizing oligopeptide transport activity. AB - To study the drug delivery to tumor by utilization of an oligopeptide transport activity, we examined the accumulation of dipeptides and the peptide-mimetic anti cancer drug, bestatin, a substrate of oligopeptide transporter PepT1. Firstly, we established HeLa cells stably expressing human peptide transporter (hPepT1) (HeLa hPepT1). Secondly, we constructed an experimental model by inoculation of HeLa hPepT1 cells subcutaneously into Balb/c nu/nu mice to demonstrate the contribution of PepT1 to the tissue-selective drug delivery. The accumulations of a hydrolysis-resistant dipeptide [(3)H]carnosine and bestatin in solid tumors formed by HeLa-hPepT1 or HeLa-pcDNA3, which are transfected with vector DNA (pcDNA3) were measured. After I.V. administration, tissue-to-plasma concentration ratios (K(p)) of both compounds, in HeLa-hPepT1 tumor was significantly greater than that of [(14)C]inulin, a marker for extracellular fluid space, those of dipeptides in muscle, or those in HeLa-pcDNA3 tumor. Furthermore, bestatin exhibited growth inhibition of HeLa-hPepT1 in vitro. In vivo, repeated oral administration of bestatin for 28 days suppressed the growth of HeLa-hPepT1 tumor specifically. When HT-1080 cells, which may naturally express oligopeptide transport activity, were transplanted, K(p) of [(3)H]carnosine was significantly increased in comparison with that in muscle. In addition, oligopeptide transport activities among various human cell lines were examined. These results provide the first demonstration for the selective delivery of oligopeptides to tumors by specific oligopeptide transport activity. PMID- 11004681 TI - Expression of SIALYL-Le(x) antigen defined by MAb AM-3 is an independent prognostic marker in colorectal carcinoma patients. AB - Expression of mucin-bound sialyl-Le(x) antigen during the progression of colorectal carcinoma and its potential prognostic value were analysed in sections of tumours from 182 patients with a documented follow-up by immunohistochemistry using the monoclonal antibody (MAb) AM-3. Two groups of colonic carcinomas with weak (n = 79) and strong (n = 103) sialyl-Le(x) expression were discerned. The percentage of strongly expressing tumours increased with the progression of the disease (UICC stage I = 10%, stage II = 46%, stage III = 63%, stage IV = 68%, p < 0.0001). Seventy-four percent of patients with carcinomas exhibiting a strong sialyl-Le(x) expression but only 34% of patients with weak sialyl-Le(x) expression died of the disease (p = 0.0026). In multivariate analysis, strong sialyl-Le(x) expression increased the relative risk of cancer-related death 3.8 fold (95% CI = 1.8-7.9, p = 0.00034). The separate analyses of patients in UICC stage II (n = 56), III (n =5 9) and IV (n = 57) revealed that strong sialyl-Le(x) expression was associated with a reduction of the 5-year overall survival rate in UICC stage II (84% vs. 54%, p = 0.0013) and in stage III patients (86% vs. 35%, p = 0.0008) after curative resection but was not relevant in patients with distant metastases. In conclusion, the strong expression of sialyl-Le(x) antigen defined by the MAb AM-3 in colorectal carcinomas is an independent unfavourable prognostic factor after curative resection in stage II and III patients. The predictive power of the sialyl-Le(x) expression may be helpful to define subgroups of patients at high risk for whom preventive adjuvant therapy can be selectively applied before the occurrence of detectable metastases. PMID- 11004682 TI - Overcoming cisplatin resistance in vitro by a free and liposome-encapsulated bile acid derivative: BAMET-R2. AB - Low water solubility and development of resistance are important drawbacks in the use of cisplatin as a cytostatic agent. A novel bile acid-cisplatin complex, Bamet-R2 [cis-diamminechlorocholylglycinateplatinum (II)], with liver vectoriality, has been synthesized. Our aim was to investigate the usefulness of this compound to overcome cisplatin resistance and to determine whether its encapsulation into liposomes increases its water solubility, uptake by liver tumor cells and cytostatic activity. Highly efficient incorporation of Bamet-R2 into liposomes permitted an increase in the concentration of the drug compared with that in the initial free solution by more than 6 x 10(6)-fold, which is 10(3)-fold higher than the encapsulation obtained for cisplatin. A partially cisplatin-resistant (87-fold) monoclonal cell line (Hepa 1-6/10R) was obtained by 2 subcloning steps of a population of mouse hepatoma Hepa 1-6 cells grown in step wise increasing cisplatin concentrations up to 10 microM. Decreased sensitivity to cisplatin was accompanied by a 3.2-fold lower drug accumulation compared to wild-type cells. Uptake was markedly increased by the binding of cisplatin to glycocholic acid in both Hepa 1-6 and Hepa 1-6/10R cells. This probably accounts for the partial overcoming (-82%) of resistance when used on Hepa 1-6/10R cells. Inclusion of Bamet-R2 into liposomes further increased the amount of the drug accumulated in both cell types and, hence, enhanced its cytostatic activity. Since both plain liposomes and Bamet-R2 have little toxicity, the formulation of this compound in liposomes may offer a substantial advantage over cisplatin in the treatment of tumors resistant to this anti-neoplastic agent. PMID- 11004683 TI - Expression of the major vault protein LRP in human non-small-cell lung cancer cells: activation by short-term exposure to antineoplastic drugs. AB - Non-small-cell lung cancer (NSCLC) cells are characterised by resistance to the toxic impact of antineoplastic drugs both in vivo and in vitro. The lung resistance-related protein (LRP), identical with the human major vault protein, is over-expressed in a variety of tumour cells characterised by intrinsic or acquired chemoresistance. We investigated the expression and cellular localisation of LRP in 16 unselected NSCLC cell lines, immortalised bronchial epithelial cells and embryonic lung fibroblasts. All cell lines analysed expressed LRP mRNA, while protein expression ranged from undetectable up to high levels. Cell fractionation and immunofluorescence staining in selected cell lines localised LRP almost exclusively to the cytoplasm. LRP was contained in the 100, 000 g pellet and absent in the soluble, cytosolic fraction and nuclei. A small proportion of LRP, however, was shown to be loosely associated with the outside of the nuclei. Sucrose gradient equilibrium centrifugation revealed presence of LRP exclusively in the fraction known to accumulate purified vault particles. Short-term exposure (16 hr) to subtoxic daunomycin (DM)-, and bleomycin (BM) concentrations significantly (up to 4-fold) enhanced LRP expression in 2/4 cell lines tested. Cisplatin (CDDP) had a minor effect while vinblastine (VBL) was ineffective. At cytotoxic conditions all drugs rather decreased than increased LRP expression. When basic LRP expression was compared with chemosensitivity, a significant correlation was detected for resistance to CDDP but not DM, doxorubicin (DOX), etoposide (VP-16), VBL and BM. Summing up, our data suggest a role of vaults both in basic CDDP resistance and, additionally, in an short-term defensive response of NSCLC cells against several other drugs. PMID- 11004684 TI - Cancer occurrence after cosmetic breast implantation in Denmark. AB - Most studies on cancer incidence after breast implantation have focused on breast cancer, while the risk of cancers at other sites has been less well investigated. We examined cancer incidence among 1,653 women who underwent cosmetic breast implant surgery at private clinics of plastic surgery in Denmark and 1,736 women attending the same clinics for other reasons during the period 1973-1995. Furthermore, we updated previously reported results among 1,114 women who received implants for cosmetic indications at public hospitals. All women were followed for cancer through the Danish Cancer Registry. In comparison with the general female population, the overall standardized incidence ratio (SIR) for cancer among women who received implants in private clinics was 1.65 [95% confidence interval (CI) = 1.17-2.27]. This elevated SIR reflected increased incidence ratios for almost all major cancer sites; however, only for non melanoma skin cancer was there an excess of more than 2 cases. No significant excess of cancer was observed among women who received implants in public hospitals (SIR = 1.10, 95% CI = 0.76-1.52) or among women attending the private clinics for other problems (SIR = 1.10, 95% CI = 0.78-1.52). The SIRs for breast cancer after breast implantation were 1.1 (95% CI = 0.5-2.2) among private clinic patients and 0.9 (95% CI = 0.4-1.7) among public hospital patients. The overall findings of these 2 implant cohorts and results from other investigations suggest that cancer risk is probably not increased among women receiving cosmetic breast implants. The inconsistent results for private clinics and public hospitals are likely related to selection bias and confounding among the private clinic patients, but our data did not permit exploration of these possibilities. Further research into the determinants of these inconsistencies is warranted. PMID- 11004685 TI - Nationwide cancer family ascertainment using Finnish Cancer Registry data on family names and places of birth for 35,761 prostate cancer patients. AB - Identification of predisposition loci to complex diseases, such as prostate cancer, requires high-quality family material, the ascertainment of which is often laborious, time-consuming and inaccurate with conventional methods. Here, we describe a new method for rapid, nationwide cancer family ascertainment using Finnish Cancer Registry data on 35,761 prostate cancer cases over a 40-year period. As members of a prostate cancer family are likely to share the same family name and place of birth, we stratified all prostate cancer cases by these 2 parameters (10,721 different names and 596 municipalities). Data were compared with the distribution of family names and places of birth for all 3.3 million Finnish men to derive standardized prevalence ratios (SPRs). A significantly elevated SPR of prostate cancer was detected for 468 (1.6%) of the 28,459 evaluable combinations of family name and place of birth. Of the 20 highest SPR values, 19 corresponded to true nuclear families, most of these having 3 or more affected cases. Two-thirds of our 50 previously established Finnish prostate cancer families were classified among this 1.6% fraction of the highest SPR values. Finally, many of the highest SPR values originated from municipalities in southern and south-western Finland. To explore whether such clusters could highlight local founder effects, we applied genealogical research to link together several families with elevated SPRs and identified an extended family with 20 prostate cancer cases with common ancestors in the early seventeenth century. In summary, a rapid novel method was developed and validated for identification of prostate cancer families from nationwide cancer registry data and for the identification of putative regional founder effects. PMID- 11004686 TI - Population based study of coffee, alcohol and tobacco use and risk of ovarian cancer. AB - Coffee, alcohol and tobacco use have been examined in many epidemiologic studies of ovarian cancer but findings have generally been inconclusive. To explain prior inconsistent findings, we sought to determine whether associations with these exposures might vary by histologic subtype of ovarian cancer or menopausal status at diagnosis. We conducted a population-based case-control study in eastern Massachusetts and New Hampshire involving 549 women with newly-diagnosed epithelial ovarian cancer and 516 control women selected either by random digit dialing or through lists of residents. Coffee and alcohol consumption was assessed through a semi-quantitative food-frequency questionnaire, and information on tobacco smoking was collected through personal interview. Consumption of coffee and caffeine was associated with increased risk for ovarian cancer but only among premenopausal women. There was no increase in risk for ovarian cancer overall associated with tobacco or alcohol use in either pre- or post-menopausal women. Association of borderline significance for tobacco and invasive serous cancers and alcohol and mucinous cancers were observed but reduced after adjustment for coffee consumption. We conclude that coffee and caffeine consumption may increase risk for ovarian cancer among premenopausal women and are findings that have some epidemiologic and biologic support. PMID- 11004687 TI - Loss of heterozygosity (LOH) at p53 is correlated with LOH at BRCA1 and BRCA2 in various human malignant tumors. PMID- 11004688 TI - Malignant hypertension - the role of the paracrine renin-angiotensin system. AB - Malignant hypertension remains one of the life-threatening complications of blood pressure elevation. It is a clinico-pathological syndrome of severe blood pressure elevation combined with malignant vascular injury. This is a characteristic form of vascular damage, with two elements: fibrinoid necrosis and endarteritis proliferans. Although the morphology of these has been well described, the molecular events are not fully understood. This review summarizes the evidence from transgenic animals for a role for the activation of a local paracrine renin-angiotensin system in the pathogenesis of malignant vascular injury. These animal models provide pathological, pharmacological, and genetic evidence supporting the hypothesis that intra-renal generation of angiotensin 2 and exposure of the microcirculation to elevated blood pressure co-operate in causing tissue damage in malignant hypertension. PMID- 11004689 TI - 'Invading edge vs. inner' (edvin) patterns of vascularization: an interplay between angiogenic and vascular survival factors defines the clinical behaviour of non-small cell lung cancer. AB - Neo-angiogenesis during neoplastic growth involves endothelial mitogenic and migration stimuli produced by cancer or tumour stromal cells. Although this active angiogenesis takes place in the tumour periphery, the process of vessel growth and survival in inner areas and its clinical role remain largely unexplored. The present study compared the microvessel score (MS) as well as the single endothelial cell score (ECS) in the invading edge and in inner areas of non-small cell lung carcinomas (NSCLCs). Three different patterns of vascular growth were distinguished: the edvin (edge vs. inner) type 1, where a low MS was observed in both peripheral and inner tumour areas; the edvin type 2, where a high MS was noted in the invading front but a low MS in inner areas; and the edvin type 3, where both peripheral and inner tumour areas had a high MS. The ECS was high in the invading edge in edvin type 2 and 3 cases and was sharply decreased in both types in inner areas, suggesting that endothelial cell migration is unlikely to contribute to the angiogenic process in areas away from the tumour front. Expression of the vascular endothelial growth factor (VEGF) and of thymidine phosphorylase (TP) was associated with a high MS in the invading edge. VEGF was associated with a high MS in inner areas (edvin 3), while TP expression was associated with edvin type 2, showing that VEGF (and not TP) contributes to the preservation of the inner vasculature. Both edvin type 2 and 3 cases showed an increased incidence of node metastasis, but edvin type 3 cases had a poorer prognosis, even in the N1-stage group. The present study suggests that tumour factors regulating angiogenesis and vascular survival are not identical. A possible method is reported to quantify these two parameters by comparing the MS in the invading edge and inner areas (edvin types). This observation may contribute to the evaluation of the effectiveness of different therapeutic approaches, namely vascular targeting vs. anti-angiogenesis. PMID- 11004690 TI - Expression of vascular endothelial growth factor by macrophages is up-regulated in poorly vascularized areas of breast carcinomas. AB - Angiogenesis is essential to the growth and metastasis of solid tumours. Vascular endothelial growth factor (VEGF) is a potent pro-angiogenic cytokine that is overexpressed in malignant tumours such as invasive carcinoma of the breast. The low oxygen tensions (hypoxia) present in these tumours are known to up-regulate the expression of VEGF by tumour cells. Human macrophages also respond to hypoxia by increasing their release of VEGF in vitro, although the effect of hypoxia on VEGF expression by macrophages in vivo has yet to be demonstrated. The present study compared the expression of VEGF by macrophages in areas of low and high vascularity in 24 invasive breast carcinomas (12 lobular, 12 ductal). The cellular distributions of VEGF protein, CD31 (vessels), and CD68 (macrophages) were compared in sequential sections for each tumour. In ten tumours, both tumour cells and macrophages were immunoreactive for VEGF protein. Use of non-isotopic in situ hybridization to localize VEGF mRNA showed that these cell types also expressed VEGF mRNA. No significant differences in the cellular distribution of VEGF protein were found between lobular and ductal carcinomas. In all tumours, macrophages accumulated in higher numbers in poorly vascularized than in highly vascularized areas. In VEGF-positive tumours, macrophages were immunoreactive for VEGF only in avascular areas where tumour cells also expressed VEGF. This suggests that VEGF expression by these two cell types may be regulated by the same microenvironmental stimuli in breast carcinomas. In addition, significantly more macrophages were present in poorly vascularized areas of VEGF-positive than VEGF-negative tumours. This suggests that VEGF may exert a chemotactic action on macrophages in vivo and guide their migration into avascular tumour sites. PMID- 11004691 TI - Expression of oestrogen receptor alpha variants in non-malignant breast and early invasive breast carcinomas. AB - Oestrogen receptor (ER) alpha variants have been described in normal breast and breast carcinomas, but their presence in a range of benign conditions and in small early invasive breast carcinomas has not been considered. Cryostat tissue sections from 19 normal and proliferative breast lesions and 44 carcinomas 15 mm and less in size detected by mammographic screening were screened for ERalpha splice variants using reverse transcriptase-nested PCR. The carcinomas were assessed for mutation by single-stranded conformational polymorphism analysis and variant forms/band shifts were sequenced. ERalpha was detected in all 19 non malignant cases and exon 7-deleted variants were found in 16 of them. Three cases showed weak expression of exon 5, and two of exon 3 variants. There was no relationship between the presence of variants and the extent of proliferative change, ER status or age. ERalpha mRNA was not detected in two carcinomas; exon 3 deletions were found in four (9. 5%) of the other carcinomas, exon 5 in two (4.8%), and exon 7 in 11 (26.2%), with two variants in four carcinomas and a total of 29.5% of all cases having detectable variants. Two point mutations were found in one, which was a tubular carcinoma. Variant forms were identified in carcinomas of all sizes (bar<10 mm) but were more frequent in those of 15 mm. There was no relationship with type, grade or receptor status. The main difference between non-malignant breast and early invasive cancers related to exons 3 and 5. The findings suggest that ERalpha variants are not involved in breast cancer development but occur with tumour progression and may be a consequence rather than a cause. PMID- 11004692 TI - Apoptosis, proliferation, and Fas (APO-1, CD95)/Fas ligand expression in medullary carcinoma of the breast. AB - Medullary carcinoma (MC) of the breast is a unique subtype of infiltrating ductal carcinoma that is characterized by a prominent lymphoid infiltrate and improved prognosis. Activated granzyme B(+)/CD8(+) cytotoxic T-lymphocytes (CTLs) infiltrating tumour cell nests constitute a major subset within the lymphoid infiltrate. As CTLs destroy target tumour cells by triggering apoptosis, it would be of interest to determine whether the apoptotic rate in MC is increased. This study evaluates the extent of apoptosis in relation to Fas (APO-1, CD95)/Fas ligand (FasL) expression in MC. Fourteen cases of typical MC (TMC) and 15 cases of atypical MC (AMC) classified according to the Ridolfi criteria, as well as 19 cases of poorly differentiated infiltrating ductal carcinoma (PDC) were evaluated. The apoptotic index (AI) was assessed by the TUNEL method on paraffin embedded tissue. Cell proliferation was evaluated immunohistochemically by PCNA staining. The level of Fas/FasL expression was determined semiquantitatively by immunohistochemistry using a four-grade scoring system. The AI was significantly increased in TMC and AMC as opposed to the PDC subgroup (2.2+/-0.8, 2.1+/-0.8, and 1.3+/-0.6, respectively; p<0.05). A significant proportion (31.8+/-7.9% in TMC and 25.8+/-9.7% in AMC) of the apoptotic tumour cells within tumour nests were in close contact with CD3(+) lymphocytes. Increased apoptosis was not accompanied by increased proliferation of tumour cells. The extent of Fas expression did not differ between the three subgroups. FasL was expressed both by tumour infiltrating lymphocytes in MC and by tumour epithelium in all three subgroups. The observation that the majority of MCs express Fas and are infiltrated by lymphocytes expressing FasL suggests that increased apoptosis in MC is mediated by Fas/FasL. However, our observation that the majority of MCs also express FasL and the fact that tumours co-expressing Fas and FasL did not show increased apoptosis suggest that there may be additional cytotoxic pathways that lead to tumour apoptosis in MC. PMID- 11004693 TI - Relationship between expression of topoisomerase II isoforms and chemosensitivity in choroidal melanoma. AB - Choroidal melanoma has a high mortality rate and responds poorly to existing chemotherapy, but unexpected ex vivo sensitivity of a subset of these tumours to topoisomerase II inhibitors has been noted. Since chemoresistance may be mediated by the molecular phenotype of tumours, immunohistochemistry has been used to study the expression of both isoforms of topoisomerase II (alpha and beta) in 29 choroidal melanomas for which chemosensitivity assay data for doxorubicin or mitoxantrone are also available. Of these, eight tumours were topoisomerase II beta-positive and 11 were topoisomerase II alpha-positive. Recent studies showing genetic abnormality (often monosomy of chromosome 3) in choroidal melanoma suggest that loss of immunostaining could be due to genomic loss rather than down regulation of topoisomerase II beta in these tumours. There was no convincing excess of anthracycline resistance in the topoisomerase II beta-negative group. Addition of topoisomerase II alpha, MDR1 (11/17 positive), LRP (16/28 positive), and MRP (5/29 positive) data in multivariate analysis did not reliably predict sensitivity or resistance. Vincristine chemosensitivity showed no relation to MDR1, LRP or MRP in 18 tumours tested. While it is possible that some tumours which do express topoisomerase II beta may respond to anthracyclines, the molecular basis of resistance or sensitivity to anthracyclines or vincristine in uveal melanoma is complex and remains incompletely understood. PMID- 11004694 TI - The restricted expression pattern of the Hodgkin's lymphoma-associated cytokine receptor CD30 is regulated by a minimal promoter. AB - One of the most peculiar immunohistological characteristics of the tumour cells of Hodgkin's lymphoma, anaplastic large cell lymphoma (ALCL), and embryonal carcinoma of the testis is the expression of the CD30 antigen. Physiologically, CD30 expression is restricted to a few activated lymphocytes in normal lymphoid tissue and a small population of decidual cells. To clarify the reasons behind this highly restricted expression pattern and to learn about the combination of transcription factors involved in this regulation in Hodgkin's lymphoma and other CD30(+) malignancies, the 5'-flanking regulatory region of the cd30 gene was analysed. The major transcription start site was determined to be 270 bases upstream of the translational start codon in the Hodgkin's lymphoma-derived cell lines L591 and L428. Reporter gene assays revealed that the CD30 promoter (-413 to 84) induces a 50- to 1000-fold higher luciferase expression in CD30(+) human lymphoid cell lines (Co, Jurkat, and the Hodgkin's lymphoma-derived cell line L540) than in CD30(-) human lymphoid cell lines (DG75, SUP-T1, and U698M), CD30( ) human carcinoma cell lines (HeLa and MCF-7), or COS1 cells. Deletion analysis defined a TATA-less, minimal promoter sequence from -164 to 84. The transcription factor Sp1 and members of the Ets family induce CD30 expression, whereas the transcription factor Sp3 diminishes its induction. These data suggest that a high Sp1/Sp3 expression ratio and a peculiar expression pattern of the Ets transcription factors are involved in the overexpression of CD30 and might contribute to the transformation of CD30(+) tumour cells. PMID- 11004695 TI - Frequent nuclear localization of ICAD and cytoplasmic co-expression of caspase-8 and caspase-3 in human lymphomas. AB - Lymphoma cells often display in vitro resistance to FAS-induced apoptosis, in which caspases act as crucial cell death effectors. Following FAS stimulation, caspase-8 activates caspase-3, which in turn activates the caspase-activated DNAse (CAD) by proteolysis of its inhibitor (ICAD). To investigate the mechanism of FAS resistance, the expression of caspase-8 was analysed by immunohistochemistry, together with that of the substrates caspase-3 and ICAD, in 52 representative samples from non Hodgkin's lymphoma (NHL), 12 from Hodgkin's disease (HD), and eight benign lymphoid tissues. In benign tissues, caspase-8 was co-expressed with caspase-3 in the cytoplasm in germinal centre (GC) cells and was co-expressed with ICAD in the nuclei of the mantle and marginal zone cells. ICAD expression was weak or absent in GC cells. Cytoplasmic staining for both caspase-8 and caspase-3 was present in 11/12 cases of diffuse large cell B-NHL. Caspase-8 positivity was nuclear and cytoplasmic in 9/9 follicular NHLs, in 5/5 mantle cell NHLs and in 6/6 marginal zone NHLs. Five out of six peripheral T-cell NHLs expressed cytoplasmic caspase-8. Ten out of the 12 HD cases lacked significant cytoplasmic staining for caspase-3 and caspase-8 in the majority of Reed-Sternberg cells. All lymphoma cases exhibited predominant nuclear ICAD positivity. Subcellular fractionation analysis of three lymphoma samples and normal mantle zone cells confirmed that ICAD and caspase-8 were at least partly localized in the nucleus. These results show that the profile of caspase-8 expression is correlated with histological lymphoma subtypes; that caspase-8 is co-expressed with caspase-3 in GC cells and their neoplastic counterparts; that ICAD has an immunohistochemical nuclear localization in vivo; and that caspase-8 and ICAD can be co-expressed in the nuclei of mantle zone and marginal zone cells; their unexpected nuclear localization allows a reappraisal of the biochemical cascade of caspase activation. PMID- 11004696 TI - In situ analysis of LKB1/STK11 mRNA expression in human normal tissues and tumours. AB - The LKB1/STK11 serine/threonine kinase is mutated in Peutz-Jeghers' syndrome and acts as a tumour suppressor. Using northern blotting and RT-PCR, LKB1 has been reported to be expressed widely in human adult tissues, although in Xenopus the expression of its homologue, XEEK1, is apparently restricted to early embryogenesis. In situ hybridization has been used to detect and localize LKB1 mRNA in a variety of adult and fetal tissues and tumours. The results show that LKB1 expression is widespread, but predominant in epithelia and in the seminiferous tubules of the testis. Expression is higher in fetal than in adult tissues. Expression also appears to be higher in many malignant tumours than in normal tissues or benign lesions, although some cancers have lost LKB1 expression, quite possibly as part of the process of tumourigenesis. These data are consistent with a widespread functional role for LKB1 in tissues of most types, and with a role for LKB1 in the pathogenesis of some sporadic cancers. LKB1 expression may primarily be related to the rate of cell replication. PMID- 11004697 TI - Molecular cytogenetic evaluation of virus-associated and non-viral hepatocellular carcinoma: analysis of 26 carcinomas and 12 concurrent dysplasias. AB - The worldwide incidence of hepatocellular carcinoma (HCC) is approximately one million cases a year. This makes HCC one of the most frequent human malignancies, especially in Asia and Africa, although the incidence is increasing also in the western world. HCC is a complication of chronic liver disease, with cirrhosis as the most important risk factor. Viral co-pathogenesis makes cirrhosis due to hepatitis B (HBV) and hepatitis C virus (HCV) infection a very important factor in the development of HCC. As curative therapy is often ruled out due to the late detection of HCC, it would be attractive to find parameters which predict malignant transformation in HBV- and HCV-infected livers. This study has used comparative genomic hybridization (CGH) to analyse 26 HCCs (11 non-viral, nine HBV, six HCV) and 12 concurrent dysplasias (five non-viral, five HBV, two HCV). Frequent gain (> or =25% of all tumours) was detected, in decreasing order of frequency, on 8q (69%), 1q (46%), 17q (46%), 12q (42%), 20q (31%), 5p (27%), 6q (27%), and Xq (27%). Frequent loss (> or =25% of all tumours) was found, in decreasing order of frequency, on 8p (58%), 16q (54%), 4q (42%), 13q (39%), 1p (35%), 4p (35%), 16p (35%), 18q (35%), 14q (31%), 17p (31%), 9p (27%), and 9q (27%). Minimal overlapping regions could be determined at multiple locations (candidate genes in parentheses). Minimal regions of overlap for deletions were assigned to 4p14-15 (PCDH7), 8p21-22 (FEZ1), 9p12-13, 13q14-31 (RB1), 14q31 (TSHR), 16p12-13.1 (GSPT1), 16q21-23 (CDH1), 17p12-13 (TP53), and 18q21-22 (DPC4, DCC). Minimal overlapping amplified sites could be seen at 8q24 (MYC), 12q15-21 (MDM2), 17q22-25 (SSTR2, GH1), and 20q12-13.2 (MYBL2, PTPN1). A single high level amplification was seen on 5q21 in an HBV-related tumour. Aberrations appeared more frequent in HBV-related HCCs than in HCV-associated tumours (p=0.008). This was most prominent with respect to losses (p=0.004), specifically loss on 4p (p=0.007), 16q (p=0.04), 17p (p=0.04), and 18q (p=0.03). In addition, loss on 17p was significantly lower in non-viral cancers than in HBV-related HCC (p<0.001). Furthermore, loss on 13q was more prevalent in HCCs in non-cirrhotic livers (p=0.02), thus suggesting a different, potentially more aggressive, pathway in neoplastic progression. A tendency (p=0.07) was observed for loss on 9q in high stage tumours; no specific changes were found in relation to tumour grade. A subset of the HCC-associated genetic changes was disclosed in the preneoplastic stage, i.e. liver cell dysplasia. This group of dysplasias showed frequent gain on 17q (25%) and frequent loss on 16q (33%), 4q (25%), and 17p (25%). The majority of the dysplasias with alterations revealed genetic changes that were also present in the primary tumour. In conclusion, firstly, this study has provided a detailed map of genomic changes occurring in HCC of viral and non viral origin, and has suggested candidate genes. Loss on 17p, including the TP53 region, appeared significantly more prevalent in HBV-associated liver cancers, whereas loss on 13q, with possible involvement of RB1, was distinguished as a possible genetic biomarker. Secondly, CGH analysis of liver cell dysplasia, both viral and non-viral, has revealed HCC-specific early genetic changes, thereby confirming its preneoplastic nature. Finally, genes residing in these early altered regions, such as CDH1 or TP53, might be associated with hepatocellular carcinogenesis. PMID- 11004698 TI - p120 expression provides a reliable indication of the rapidity of cell duplication in cancer cells independently of tumour origin. AB - p120 is a nucleolar protein that has been immunocytochemically detected in rapidly proliferating cells of a variety of human malignancies. In the present study, the relationship between p120 expression and the rapidity of cell proliferation was evaluated in 48 human tumours of different origins. The cell proliferation rate of cancer cells was determined by quantitative analysis of AgNOR proteins. p120 immunostaining and AgNOR protein quantity were measured by image cytometry and a highly significant correlation was found between the two variables, as evaluated by linear regression analysis (r=0.98, p<0.0001). The relationship between p120 expression and the rapidity of cell duplication was also studied in vitro, in six human cancer cell lines derived from different tumour types, characterized by various doubling times (ranging from 20 to 77 h). p120 expression was determined on western blots using specific anti-p120 monoclonal antibodies. Densitometric analysis revealed a highly significant inverse correlation between the integrated optical density values of the chemoluminescence bands at 120 kD and the cell line doubling times (r=-0.93; p=0.007). The same result was obtained in situ by correlating p120 immunostaining of the cytological preparations obtained from the six cancer cell lines with the corresponding doubling time (r=-0.98, p<0.0001). These results indicate that in cancer cells, the quantitative expression of p120 is directly related to the rapidity of cell duplication, independently of the tumour origin. PMID- 11004699 TI - Clonal patterns in phaeochromocytomas and MEN-2A adrenal medullary hyperplasias: histological and kinetic correlates. AB - The relationship among histological features, cell kinetics, and clonality has not been studied in adrenal medullary hyperplasias (AMHs) and phaeochromocytomas (PCCs). Thirty-four PCCs (23 sporadic and 11 MEN-2A (multiple endocrine neoplasia type 2A)-related tumours, the latter associated with AMH) from females were included in this study. Representative samples were histologically evaluated and microdissected to extract DNA and evaluate the methylation pattern of the androgen receptor alleles. At least two tissue samples (from the peripheral and internal zones in each tumour) were analysed with appropriate tissue controls run in every case. The same areas were selected for MIB-1 staining and in situ end labelling (ISEL). Malignant PCCs were defined by histologically confirmed distant metastases. All monoclonal AMH nodules from the same patient showed the same X chromosome inactivated. Six sporadic PCCs revealed liver metastases (malignant PCC) and eight additional sporadic PCCs showed periadrenal infiltration (locally invasive PCC). All informative PCCs were monoclonal, except for five locally invasive PCCs and one benign PCC that revealed polyclonal patterns. Those cases also showed a fibroblastic stromal reaction with prominent blood vessels, focal smooth muscle differentiation, and significantly higher MIB-1 (126.8+/-29.9) and ISEL (50.9+/-12.8) indices. Concordant X-chromosome inactivation in nodules from a given patient suggests that MEN-2A AMH is a multifocal monoclonal condition. A subgroup of PCCs characterized by balanced methylation of androgen receptor alleles, high cellular turnover, and stromal proliferation also shows locally invasive features. PMID- 11004700 TI - Errors in histological grading by prostatic needle biopsy specimens: frequency and predisposing factors. AB - Sampling error is an inherent problem of prostate biopsy. Consequently, there are problems in determining whether a given carcinoma is clinically significant on the basis of biopsy results. This study assesses the factors that predispose to errors in biopsy grading, as well as the dimensions of sampling error due to these factors. Among 187 cases, biopsy grading error was retrospectively related to grade heterogeneity in the prostate and to biopsy-related factors. Clinically relevant biopsy grading errors occurred in a quarter of the cases. Of all grading errors, at least 17% resulted from misinterpretation by the pathologist only. Overall, prostates with grade heterogeneity revealed grading errors twice as frequently as specimens without grade heterogeneity. In most cases, however, grading error resulted from multiple factors, such as the number and length of cores obtained (p<0.05). This was an important finding because the mean core length was only 9.4 mm, whereas the biopsy needle is designed to obtain cores of 15 mm. Moreover, clinically relevant biopsy grading error had occurred in almost half of the cases when the Gleason score was based on a tumour deposit measvring less than 0.5 mm (p<0.05). The clinical consequences of these findings are important. Clinicians should try to obtain at least six biopsies, each 15 mm in length, to minimize grading error. Pathologists should be cautious in reporting Gleason scores based on tumour lesions smaller than 400x total magnification field. Interpretation could be refined, when necessary, by warning the urologist of the Limitations of the biopsy report. PMID- 11004701 TI - Expression of angiogenic factor thymidine phosphorylase and angiogenesis in human atherosclerosis. AB - In atherosclerosis, leukocyte migration into the plaque is thought to occur across the endothelium of the arterial lumen. However, intraplaque microvessels have been noted. While the significance of, and stimuli for these are uncertain, it seems possible that they may represent a second portal of entry for leukocytes into the plaque. This study performed a basic characterization of intraplaque microvessels and tested the hypothesis that the novel angiogenic factor thymidine phosphorylase (TP) is expressed in atherosclerosis. Immunocytochemistry was performed on aortic and coronary plaques and morphometry on coronary plaques. In plaques from both sites, macrophages, foam cells, and giant cells were immunoreactive for the angiogenic factors TP and vascular endothelial growth factor. Venule-like intraplaque microvessels expressed endothelial leukocyte adhesion molecules HLA-DR and ICAM-1, in contrast to the endothelium overlying the plaque. In coronary plaques, there was a correlation between severity of stenosis and plaque microvascular density. These results are consistent with a role for plaque macrophage/foam cell TP in stimulating plaque angiogenesis. While attention has focused on dysfunction of the endothelium overlying the plaque, microvascular endothelium may also represent a portal of entry for leukocytes into established plaques. PMID- 11004702 TI - The relationship between persistent secretion of RANTES and residual infiltration of eosinophils and memory T lymphocytes after Helicobacter pylori eradication. AB - Helicobacter pylori (HP)-infected gastric mucosa displays a conspicuous infiltration of mononuclear cells as well as neutrophils. RANTES is a potent chemoattractant peptide for memory T lymphocytes and eosinophils. RANTES protein concentration and the numbers of RANTES-, CD45RO-, and major basic protein (MBP) positive cells were therefore evaluated in the gastric mucosa from 51 patients with HP-positive chronic gastritis before and after HP eradication and from 22 HP negative healthy volunteers. RANTES protein concentration was significantly elevated in HP-positive cases and remained high after HP eradication. The numbers of RANTES-, CD45RO-, and MBP-positive cells were significantly increased in HP positive cases and were well correlated with RANTES protein levels. All tended to decrease after HP eradication, but did not reach the level of HP-negative cases, even at 24 months after HP eradication. It was concluded that persistent expression and secretion of RANTES were closely related to residual infiltration of memory T lymphocytes and eosinophils, for a prolonged period after HP eradication. This seems to be an important mechanism of prolonged gastric mucosal immune response against HP infection, even after HP eradication, and of persistent mucosal damage and atrophy. PMID- 11004703 TI - Increased expression of cytoplasmic tail-containing form of gp600/megalin in active Heymann nephritis. AB - Active Heymann nephritis of rat, an autoimmune glomerular disease, is a model of human membranous glomerulonephropathy. The major target autoantigen of this disease is a 600 kD renal glycoprotein, variously named gp600/gp330/megalin/LRP2. It has been shown previously that there is a marked increase in the gp600/megalin mRNA level in glomeruli of rats with this disease. A rabbit antibody was prepared against a peptide located at the very end of the cytoplasmic tail of rat gp600/megalin. Affinity purified anti-peptide antibody was monospecific and reacted only with gp600/megalin on the immunoblots of total kidney proteins. Immunofluorescence microscopy on frozen kidney sections probed with this antibody revealed the presence of the cytoplasmic tail-containing form of gp600/megalin in both proximal tubules and glomeruli. Expression in glomeruli of normal kidneys was less than in tubules but was clearly detectable. In active Heymann nephritis kidneys, expression of this form was increased most remarkably in glomeruli. These results, showing increased expression of the cytoplasmic tail-containing form in active Heymann nephritis glomeruli, contradict previous reports of its absence. This is the first study to show that there is increased expression of the cytoplasmic tail-containing form of gp600/megalin in the glomeruli of rats with this disease and its role in active Heymann nephritis is discussed. PMID- 11004704 TI - The effect of environmental cues on the differentiation of myofibroblasts in peritoneal granulation tissue. AB - This study investigated the effect of haemodynamic stress, active stretch, and neuronal input on the differentiation of myofibroblasts in peritoneal granulation tissue. Lengths of silastic tubing (10 mm long x 3 mm diameter) were placed in the peritoneal cavity of the rat. By 2 weeks, a capsule of granulation tissue had formed around the tubing. This capsule consisted of several layers of myofibroblasts and the matrix that they had produced, overlaid by a single layer of mesothelial cells. The silastic tubing was removed and at the same time, the living tube of tissue was everted so that the mesothelium now lined its inner surface. To examine the effect of haemodynamic factors on myofibroblast differentiation, the 10 mm long tubes of mesothelial-lined granulation tissue were transplanted into the severed abdominal aorta of the same rat in which the granulation tissue was grown. End-to-end anastomoses were performed to extend the existing aorta. At 1, 2, and 3 months post-transplantation, the grafts were removed and a progressive increase in the percent volume fraction of myofilaments (% V(v)myo) was observed (from 35.7+/-1.6% to 58.7 3+/-1.4%; p<0.05). To determine whether the active stretching that occurs in vivo could account for differentiation of the constituent myofibroblasts, tubes of granulation tissue were placed into a mechanical device in which they underwent continuous stretching of 5-10% elongation from the resting position at 50 cycles per minute for 3, 24 or 72 h. This caused a significant (p<0. 05) increase in %V(v)myo after 72 h. Granulation tissue was also transplanted into the rat anterior eye chamber, where it became surrounded by adrenergic nerves supplying the host iris. Two months after implantation, there was no significant change in the %V(v)myo of the myofibroblasts (35.7+/-1.6% to 33.3+/-2.7%). These studies show that myofibroblasts of the granulation tissue encapsulating free-floating foreign bodies in the peritoneal cavity further differentiate towards a smooth muscle phenotype when transplanted into a smooth muscle environment, namely the abdominal aorta. Similar changes are seen when the granulation tissue is subjected to active, intermittent stretch in vitro, while the presence of nerves has no effect. PMID- 11004706 TI - [The previous law concerning infectious diseases and the new infectious diseases control law - the problems for the enforcement of the new infectious diseases control law in medical situation]. PMID- 11004705 TI - Role of macrophage scavenger receptor in endotoxin shock. AB - Lipopolysaccharide (LPS) is known to bind to several receptors on macrophages, including CD14 and macrophage scavenger receptor class A types I and II (MSR-A), and stimulates macrophages to release various inflammatory mediators. MSR-A recognizes a broad range of polyanionic ligands such as chemically modified lipoproteins, LPS of Gram-negative bacteria, and lipoteichoic acid of Gram positive bacteria, suggesting a role in host defence. In this study, mice lacking MSR-A were used to elucidate the role of MSR-A in endotoxin shock. Peritoneal macrophages from MSR-A-deficient (MSR-A(-/-)) mice bound less remarkably to LPS than those from wild-type (MSR-A(+/+)) mice and the binding activity of MSR A(+/+) macrophages to LPS was reduced by the addition of an anti-MSR-A antibody. Clearance of LPS in serum was retarded in MSR-A(-/-) mice after intraperitoneal administration of LPS. LPS-induced expression of cytokines in the liver was similar in MSR-A(+/+) and MSR-A(-/-) mice, but levels of interleukin (IL)-1beta expression and serum IL-1beta were lower in MSR-A(-/-) mice. Administration of large doses of LPS resulted in a higher mortality of MSR-A(+/+) mice and pretreatment with an IL-1 receptor antagonist reduced the mortality. Thus, MSR-A mediated macrophage activation plays a negative role in protecting mice from endotoxin shock by enhancing IL-1beta production by macrophages. PMID- 11004707 TI - [Comparative evaluation of BACTEC MGIT 960 system with MB/BacT and egg-based media for recovery of mycobacteria]. AB - Fully automated, nonradiometric mycobacteria culture systems, BACTEC MGIT 960 (Becton Dickinson Microbiology Systems, Sparks, Md, U.S.A.) and MB/BacT (Organon Teknika, Durham, NC, U.S.A.) were evaluated in comparison with three different eggbased media (3% Ogawa, Ogawa K, and Vite) for the ability to detect mycobacteria in clinical sputum specimens. Sputum specimens were processed by semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) for the automated systems, and by cetylpyridium chloride-succinic acid-NaCl for the egg-based media. A total of 954 sputum specimens were processed, and the recovery of mycobacteria by the BACTEC MGIT 960 was performed in a commercial laboratory. Overall, the frequency of breakthrough contamination was <1% for the three egg based media, ranging from 0. 42% to 0.63%. Whereas, the frequency of false positives due to breakthrough contamination was 1.89% for MB/BacT and 20.1% for BACTEC MGIT 960. A total of 237 isolates of Mycobacterium tuberculosis complex and 167 isolates of nontuberculous mycobacteria (NTM) were recovered. The highest recovery ratio was obtained by MB/BacT (95.8%), followed by the egg-based media, Vite (74.3%), Ogawa K (65.8%), and 3% Ogawa (58.9%). The recovery ratio by BACTEC MGIT 960 was the lowest, and estimated at 43.1%, mainly due to a high frequency of breakthrough contamination. However, even after omission of these false positives reported by BACTEC MGIT 960, the recovery ratio by this system was comparable to that of 3% Ogawa media. The time to detection of 50% of positive cultures of M. tuberculosis complex by BACTEC MGIT 960 and MB/BacT was 20 days and 17 days, respectively.>From these results, it may be concluded that MB/BacT is superior to BACTEC MGIT 960 and egg-based media for the recovery of mycobacteria from sputum specimens. Furthermore, based on the outcome of this study, we think that considerable improvements are necessary for the clinical application of BACTEC MGIT 960. These improvements should particularly be focused on reducing the false positive ratio caused by contamination, and culture media, which effectively support mycobacterial growth. PMID- 11004708 TI - [Evaluation of "Helicobacter-Selective-Medium (Nissui Seiyaku)" for rapid detection of Helicobacter pylori]. AB - We evaluated Helicobacter Selective Agar (HSA) medium (Nissui Pharmaceuticals Co., Ltd., Tokyo, Japan), which has been newly developed and has been commercially available since June in 1998. HSA medium strongly inhibited the growth of possible contaminants from gastric biopsy-specimens, such as alpha streptococci, Neisseria species, Enterococcus gallinarum, Pseudomonas aeruginosa, Corynebacterium species, Capnocytophaga species, and Candida albicans. Cultures run in parallel with Helicobacter Pylori Agar (HPA) medium (Eiken Chemical Co., Ltd., Tokyo, Japan) disclosed no discordance in the detective rates of H. pylori strains between the media examined. Inaddition, the numbers of the colonies and the colony-sizes grown on the plates were also in good agreement with each other. However, it would be strikingly favorable that the appearing colonies on the HSA medium turned purple. This coloration enabled us to detect the emergence of the colonies much earlier and easier when compared with the conventional other media including HPA medium (Eiken). These findings led us to conclude that HSA medium (Nissui) was superior to HPA medium (Eiken) in the prompt detection and the rapid identification of H. pylori in routine clinical microbiology. PMID- 11004709 TI - Preventive procedures against GBS infection by means of antibody measurement. AB - OBJECTIVE: Screening group B Streptococcus (GBS) in the vagina of pregnant women and measuring serum level of its type-specific antibody would be useful for cost benefit of the prevention against GBS infection. STUDY DESIGN: The subjects included a total of 1,150 pregnant women who consented to the study. Serotypes of GBS detected were classified with antiserum. Serum type-specific antibody titers were measured by a bacterial agglutination method. RESULTS: Of a total of 1,150 pregnant women, 250 cases (21.7%) had GBS in the vagina. The turn of GBS serotype occurrence was types VI (NT6) (27. 2%), VIII (JM9) (25.2%), III (11.2%), Ia (8.8%), and Ib (8.0%). None or low type-specific antibody titer was 41.0% of Ia, 20.0% of Ib, 22. 0% of II, 15.0% of III, 65.0% of VI, and 69.0% of VIII incarriers. Noneonatal GBS infection occurred under the empirically treatment. CONCLUSION: The measurement of serum type-specific antibody against GBS would be informative for the cost-benefit treatment of the vaginal GBS in pregnant women. PMID- 11004710 TI - Basic studies on group B streptococcal (GBS) culture medium. AB - Yeast extract and Serum are said to be necessary for the pigmentation of Group B Streptococcus (hereafter, GBS). We're examined the necessity of these for the GBS pigmentation. Firstly, we examined the following four types of culture media. (1) The medium in which serum was excluded from the New Granada medium (hereafter, called Basic medium). (2) New Granada medium (hereafter, S medium). (3) The medium in which Yeast extract was added to (1) (hereafter, Y medium). (4) The medium in which added Serum and Yeast extract were added to (1) (hereafter, SY medium). As for GBS tested (7 serotypes, 27 strains), Basic medium showed the strongest pigmentation potency, but, S and SY medium showed the weakest. About the quickness of pigmentation initiation, similar to the potency above, Basic medium was the quickest, followed by Y medium, and then S medium and SY medium. Among the media examined, there were no differences in the relationships between the quantity of GBS applied and the positive ratio of each media.>From these results, among four types of the media, Basic medium is thought to be the best one for the GBS detection. PMID- 11004711 TI - [Application of DNA-DNA hybridization, DDH MYCOBACTERIA 'Kyokuto' to species identification of mycobacteria grown in Middlebrook 7H9 broth]. AB - A colorimetric DNA-DNA hybridization for the genetic identification of mycobacteria, DDH MYCOBACTERIA 'KYOKUTO' (Kyokuto Pharmaceuticals, Tokyo) was evaluated for the clinical isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT (Organon Teknika, Durham, NC, U.S.A.). When the MB/BacT gave the positive interpretation, 10 ml of Middlebrook 7H9 broth was collected from the bottle. After centrifugation at 3,000 rpm for 10 min, two drops of acetone were added to the pellet, then let it stand for one hour at the room temperature. The air-dried pellet was resuspended in a small volume of distilled water, and proceeded to the identification described. Of 136 clinical isolates of mycobacteria, comprising of 76 M. tuberculosis complex and 60 nontuberculous mycobacteria (NTM), ninety-five (70%) were correctly identified when compared to the reference identification. Thirty (22%) isolates resulted in the unidentified due to negative reaction throughout the test wells, and the remaining 11 (8%) were also unidentified due to low likelihoods. According to the package insert, the DDH MYCOBACTERIA may not be applicable to the isolates grown in Middlebrook broth. However, our test procedure using acetone prior to the extraction of bacterial DNA enables us to directly identify the isolates of mycobacteria grown in Middlebrook 7H9 broth of MB/BacT. PMID- 11004712 TI - Role of the hypothalamic pituitary adrenal axis in the control of the response to stress and infection. AB - The release of adrenocorticotropin (ACTH) from the corticotrophs is controlled principally by vasopressin and corticotropin-releasing hormone (CRH). Oxytocin may augment the release of ACTH under certain conditions, whereas atrial natriuretic peptide acts as a corticotropin release-inhibiting factor to inhibit ACTH release by direct action on the pituitary. Glucocorticoids act on their receptors within the hypothalamus and anterior pituitary gland to suppress the release of vasopressin and CRH and the release of ACTH in response to these neuropeptides. CRH neurons in the paraventricular nucleus also project to the cerebral cortex and subcortical regions and to the locus ceruleus (LC) in the brain stem. Cortical influences via the limbic system and possibly the LC augment CRH release during emotional stress, whereas peripheral input by pain and other sensory impulses to the LC causes stimulation of the noradrenergic neurons located there that project their axons to the CRH neurons stimulating them by alpha-adrenergic receptors. A muscarinic cholinergic receptor is interposed between the alpha-receptors and nitric oxidergic interneurons which release nitric oxide that activates CRH release by activation of cyclic guanosine monophosphate, cyclooxygenase, lipoxygenase and epoxygenase. Vasopressin release during stress may be similarly mediated. Vasopressin augments the release of CRH from the hypothalamus and also augments the action of CRH on the pituitary. CRH exerts a positive ultrashort loop feedback to stimulate its own release during stress, possibly by stimulating the LC noradrenergic neurons whose axons project to the paraventricular nucleus to augment the release of CRH. PMID- 11004713 TI - Proliferative signaling initiated in ACTH receptors. AB - This article reviews recent results of studies aiming to elucidate modes of integrating signals initiated in ACTH receptors and FGF2 receptors, within the network system of signal transduction found in Y1 adrenocortical cells. These modes of signal integration should be central to the mechanisms underlying the regulation of the G0-->G1-->S transition in the adrenal cell cycle. FGF2 elicits a strong mitogenic response in G0/G1-arrested Y1 adrenocortical cells, that includes a) rapid and transient activation of extracellular signal-regulated kinases-mitogen-activated protein kinases (ERK-MAPK) (2 to 10 min), b) transcription activation of c-fos, c-jun and c-myc genes (10 to 30 min), c) induction of c-Fos and c-Myc proteins by 1 h and cyclin D1 protein by 5 h, and d) onset of DNA synthesis stimulation within 8 h. ACTH, itself a weak mitogen, interacts with FGF2 in a complex manner, blocking the FGF2 mitogenic response during the early and middle G1 phase, keeping ERK-MAPK activation and c-Fos and cyclin D1 induction at maximal levels, but post-transcriptionally inhibiting c Myc expression. c-Fos and c-Jun proteins are mediators in both the strong and the weak mitogenic responses respectively triggered by FGF2 and ACTH. Induction of c Fos and stimulation of DNA synthesis by ACTH are independent of PKA and are inhibited by the PKC inhibitor GF109203X. In addition, ACTH is a poor activator of ERK-MAPK, but c-Fos induction and DNA synthesis stimulation by ACTH are strongly inhibited by the inhibitor of MEK1 PD98059. PMID- 11004714 TI - The neuroimmune-endocrine axis: pathophysiological implications for the central nervous system cytokines and hypothalamus-pituitary-adrenal hormone dynamics. AB - Cytokines are molecules that were initially discovered in the immune system as mediators of communication between various types of immune cells. However, it soon became evident that cytokines exert profound effects on key functions of the central nervous system, such as food intake, fever, neuroendocrine regulation, long-term potentiation, and behavior. In the 80's and 90's our group and others discovered that the genes encoding various cytokines and their receptors are expressed in vascular, glial, and neuronal structures of the adult brain. Most cytokines act through cell surface receptors that have one transmembrane domain and which transduce a signal through the JAK/STAT pathway. Of particular physiological and pathophysiological relevance is the fact that cytokines are potent regulators of hypothalamic neuropeptidergic systems that maintain neuroendocrine homeostasis and which regulate the body's response to stress. The mechanisms by which cytokine signaling affects the function of stress-related neuroendocrine systems are reviewed in this article. PMID- 11004715 TI - Familial hyperaldosteronism. AB - Aldosterone, the major circulating mineralocorticoid, participates in blood volume and serum potassium homeostasis. Primary aldosteronism is a disorder characterised by hypertension and hypokalaemia due to autonomous aldosterone secretion from the adrenocortical zona glomerulosa. Improved screening techniques, particularly application of the plasma aldosterone:plasma renin activity ratio, have led to a suggestion that primary aldosteronism may be more common than previously appreciated among adults with hypertension. Glucocorticoid remediable aldosteronism (GRA) was the first described familial form of hyperaldosteronism. The disorder is characterised by aldosterone secretory function regulated chronically by ACTH. Hence, aldosterone hypersecretion can be suppressed, on a sustained basis, by exogenous glucocorticoids such as dexamethasone in physiologic range doses. This autosomal dominant disorder has been shown to be caused by a hybrid gene mutation formed by a crossover of genetic material between the ACTH-responsive regulatory portion of the 11ss hydroxylase (CYP11B1) gene and the coding region of the aldosterone synthase (CYP11B2) gene. Familial hyperaldosteronism type II (FH-II), so named to distinguish the disorder from GRA or familial hyperaldosteronism type I (FH-I), is characterised by autosomal dominant inheritance of autonomous aldosterone hypersecretion which is not suppressible by dexamethasone. Linkage analysis in a single large kindred, and direct mutation screening, has shown that this disorder is unrelated to mutations in the genes for aldosterone synthase or the angiotensin II receptor. The precise genetic cause of FH-II remains to be elucidated. PMID- 11004716 TI - Plasma metanephrines: a novel and cost-effective test for pheochromocytoma. AB - Pheochromocytomas are rare chromaffin cell tumors that nevertheless must be excluded in large numbers of patients who develop sustained or episodic hypertension as well as in many others with suggestive symptoms or with a familial history of pheochromocytoma. Diagnosis of pheochromocytoma depends importantly on biochemical evidence of excess catecholamine production by a tumor. Imperfect sensitivity and specificity of commonly available biochemical tests and the low incidence of the tumor among the tested population mean that considerable time and effort can be expended in confirming or ruling out pheochromocytoma in patients where the tumor is suspected. Measurements of plasma free metanephrines provide a superior test compared to other available tests for diagnosis of pheochromocytoma. In particular, the high sensitivity of plasma free metanephrines means that a normal test result reliably excludes all but the smallest of pheochromocytomas so that no other tests are necessary. Measurements of plasma free metanephrines, when systematically combined with other diagnostic procedures outlined in this review, provide a more efficient, reliable and cost effective approach for diagnosis of pheochromocytoma than offered by previously available approaches. PMID- 11004718 TI - Adrenal incidentaloma. AB - Incidentally discovered adrenal masses, or adrenal incidentalomas, have become a common clinical problem owing to wide application of radiologic imaging techniques. This definition encompasses a heterogeneous spectrum of pathologic entities, including primary adrenocortical and medullary tumors, benign or malignant lesions, hormonally active or inactive lesions, metastases, and infections. Once an adrenal mass is detected, the clinician needs to address two crucial questions: is the mass malignant, and is it hormonally active? This article provides an overview of the diagnostic clinical approach and management of the adrenal incidentaloma. Mass size is the most reliable variable to distinguish benign and malignant adrenal masses. Adrenalectomy should be recommended for masses greater than 4.0 cm because of the increased risk of malignancy. Adrenal scintigraphy has proved useful in discriminating between benign and malignant lesions. Finally, fine-needle aspiration biopsy is an important tool in the evaluation of oncological patients and it may be useful in establishing the presence of metastatic disease. The majority of adrenal incidentalomas are non-hypersecretory cortical adenomas but an endocrine evaluation can lead to the identification of a significant number of cases with subclinical Cushing's syndrome (5-15%), pheochromocytoma (1.5-13%) and aldosteronoma (0-7%). The first step of hormonal screening should include an overnight low dose dexamethasone suppression test, the measure of urinary catecholamines or metanephrines, serum potassium and, in hypertensive patients, upright plasma aldosterone/plasma renin activity ratio. Dehydroepiandrosterone sulfate measurement may show evidence of adrenal androgen excess. PMID- 11004717 TI - Salivary cortisol as a tool for physiological studies and diagnostic strategies. AB - Salivary cortisol is an index of plasma free cortisol and is obtained by a noninvasive procedure. We have been using salivary cortisol as a tool for physiological and diagnostic studies, among them the emergence of circadian rhythm in preterm and term infants. The salivary cortisol circadian rhythm in term and premature infants was established between 8 and 12 postnatal weeks. In the preterm infants the emergence of circadian rhythm was parallel to the onset of sleep rhythm. We also studied the use of salivary cortisol for screening for Cushing's syndrome (CS) in control and obese outpatients based on circadian rhythm and the overnight 1 mg dexamethasone (DEX) suppression test. Salivary cortisol was suppressed to less than 100 ng/dl after 1 mg DEX in control and obese patients. A single salivary cortisol measurement at 23:00 h and again after 1 mg DEX above the 90th percentile of the obese group values had sensitivity and specificity of 93 and 93% (23:00 h), and 91 and 94% (after DEX), respectively. The sensitivity improved to 100% when we combined both parameters. We also studied 11 CS children and 21 age-matched primary obese children for whom salivary cortisol sensitivity and specificity were 100/95% (23:00 h), and 100/95% (1 mg DEX), respectively. Similar to adults, sensitivity and specificity of 100% were obtained by combining 23:00 h and 1 mg DEX. The measurement of salivary cortisol is a useful tool for physiological studies and for the diagnosis of CS in children and adults on an outpatient basis. PMID- 11004719 TI - Clinical results of the use of mitotane for adrenocortical carcinoma. AB - Mitotane (o,p'-DDD) acts mainly as an inhibitor of intramitochondrial pregnenolone and cortisol synthesis. Its adrenolytic effect depends on metabolic activation due to conversion to o,p'-DDA and o,p'-DDE. The drug has been used for 40 years in the treatment of adrenocortical carcinoma, mainly its regional and metastatic stage, as an adjuvant to surgical resection of the tumor. In the medical literature there are controversial opinions about its efficacy for the treatment of adrenocortical carcinoma. In our experience, mitotane administered immediately after surgery appeared to be much more efficient than when administered later. We have administered this drug in all cases of microscopically confirmed adrenocortical carcinoma, irrespectively of stage at the time of surgery, for fear of a false too optimistic classification. In our series of 82 patients with adrenocortical carcinoma, 59 patients have been treated with mitotane, 32 of them immediately after surgery, and 27 with a delay of 2 to 24 months. Today there are 18 survivors in the group of patients treated with mitotane soon after the operation and only 6 survivors in the group receiving mitotane with a delay. All patients were simultaneously given replacement therapy. Undesired effects of mitotane administration included increased aminotransferase and alkaline phosphatase activity, decreased white cell, platelet or red cell number, and myasthenia. Furthermore, we used mitotane with good results in Cushing's syndrome of non-malignant origin as pre-treatment before surgery or in long-term treatment for patients with poor tolerance of other adrenal inhibitors. PMID- 11004720 TI - Conventional and novel strategies in the treatment of adrenocortical cancer. AB - Adrenocortical carcinoma is a highly malignant neoplasm with an incidence of two per million people per year. Several treatment strategies have resulted in temporary or partial tumor regression but very few cases have attained long survival. Surgical resection of the primary tumor and metastases is most effective. Several chemotherapeutic protocols have been employed with variable success. Mitotane (o,p'-DDD) is an adrenalytic drug effective in inducing a tumor response in 33% of patients treated. Mitotane requires metabolic transformation for therapeutic action. Tumors may vary in their ability to metabolize mitotane and the ability of tumors to transform mitotane may predict the clinical response to the drug. Preliminary data show a possible correlation between metabolic activity of neoplastic adrenocortical tissue and response to mitotane. We have attempted to develop mitotane analogs with enhanced adrenalytic effect. Compared to mitotane, a di-chloro compound, the bromo-chloro and di-bromo analogs appear to have a greater effect. Future approaches to the treatment of adrenocortical carcinoma are likely to be based on blocking or reversing the biological mechanisms of tumorigenesis. Angiogenic and chemotactic mechanisms may play a role in adrenal tumor growth and inhibition of these mechanisms may result in inhibition of tumor growth. New mitotane analogs with greater adrenalytic potential could be a promising approach to developing more effective and selective therapies for adrenal cancer. Alternative approaches should attempt to suppress tumor growth by means of compounds with anti-angiogenic and anti chemotactic activity. PMID- 11004721 TI - The diversity of abnormal hormone receptors in adrenal Cushing's syndrome allows novel pharmacological therapies. AB - Recent studies from several groups have indicated that abnormal or ectopic expression and function of adrenal receptors for various hormones may regulate cortisol production in ACTH-independent hypercortisolism. Gastric inhibitory polypeptide (GIP)-dependent Cushing's syndrome has been described in patients with either unilateral adenoma or bilateral macronodular adrenal hyperplasia; this syndrome results from the large adrenal overexpression of the GIP receptor without any activating mutation. We have conducted a systematic in vivo evaluation of patients with adrenal Cushing's syndrome in order to identify the presence of abnormal hormone receptors. In macronodular adrenal hyperplasia, we have identified, in addition to GIP-dependent Cushing's syndrome, other patients in whom cortisol production was regulated abnormally by vasopressin, ss adrenergic receptor agonists, hCG/LH, or serotonin 5HT-4 receptor agonists. In patients with unilateral adrenal adenoma, the abnormal expression or function of GIP or vasopressin receptor has been found, but the presence of ectopic or abnormal hormone receptors appears to be less prevalent than in macronodular adrenal hyperplasia. The identification of the presence of an abnormal adrenal receptor offers the possibility of a new pharmacological approach to control hypercortisolism by suppressing the endogenous ligands or by using specific antagonists for the abnormal receptors. PMID- 11004722 TI - 21-Hydroxylase deficiency in Brazil. AB - We determined the frequency of large rearrangements and point mutations in 130 Brazilian patients with 21-hydroxylase deficiency and correlated genotype with phenotype. The frequency of CYP21 deletions was lower (4.4%) than in most of the previous series described, whereas the frequency of large gene conversions was similar to the frequency reported in the literature (6.6%). The most frequent point mutations were I2 splice (41.8% in salt wasting - SW), I172N (32.6% in simple virilizing - SV) and V281L (40.2% in the late onset form - LO). The frequency of the nine most common point mutations was similar to that reported for other countries. The 93 fully genotyped patients were classified into 3 mutation groups based on the degree of enzymatic activity (A<2%, B approximately 2%, C>20%). In group A, 62% of cases presented the SW form; in group B, 96% the SV form, and in group C, 88% the LO form. We diagnosed 80% of the affected alleles after screening for large rearrangements and 15 point mutations. To diagnose these remaining alleles we sequenced the CYP21 gene of one patient with the SV form and identified a heterozygous G-->A transition in codon 424. This mutation leads to a substitution of glycine by serine in a conserved region and was also found in a compound heterozygous state in 4 other patients. The mutation G424S presented a linkage disequilibrium with CYP21P and C4A gene deletions and HLA DR17, suggesting a probable founder effect. Search for the G424S mutation in other populations will reveal if it is restricted to the Brazilian patients or if it has a wider ethnic distribution. PMID- 11004723 TI - Amplification of 9q34 in childhood adrenocortical tumors: a specific feature unrelated to ethnic origin or living conditions. AB - Adrenocortical tumors (ACT) in children under 15 years of age exhibit some clinical and biological features distinct from ACT in adults. Cell proliferation, hypertrophy and cell death in adrenal cortex during the last months of gestation and the immediate postnatal period seem to be critical for the origin of ACT in children. Studies with large numbers of patients with childhood ACT have indicated a median age at diagnosis of about 4 years. In our institution, the median age was 3 years and 5 months, while the median age for first signs and symptoms was 2 years and 5 months (N = 72). Using the comparative genomic hybridization technique, we have reported a high frequency of 9q34 amplification in adenomas and carcinomas. This finding has been confirmed more recently by investigators in England. The lower socioeconomic status, the distinctive ethnic groups and all the regional differences in Southern Brazil in relation to patients in England indicate that these differences are not important to determine 9q34 amplification. Candidate amplified genes mapped to this locus are currently being investigated and Southern blot results obtained so far have discarded amplification of the abl oncogene. Amplification of 9q34 has not been found to be related to tumor size, staging, or malignant histopathological features, nor does it seem to be responsible for the higher incidence of ACT observed in Southern Brazil, but could be related to an ACT from embryonic origin. PMID- 11004724 TI - Adrenocortical tumors in children. AB - Childhood adrenocortical tumors (ACT) are rare. In the USA, only about 25 new cases occur each year. In Southern Brazil, however, approximately 10 times that many cases are diagnosed each year. Most cases occur in the contiguous states of Sao Paulo and Parana. The cause of this higher rate has not been identified. Familial genetic predisposition to cancer (p53 mutations) and selected genetic syndromes (Beckwith-Wiedemann syndrome) have been associated with childhood ACT in general but not with the Brazilian counterpart. Most of the affected children are young girls with classic endocrine syndromes (virilizing and/or Cushing). Levels of urinary 17-ketosteroids and plasma dehydroepiandrosterone sulfate (DHEA S), which are abnormal in approximately 90% of the cases, provide the pivotal clue to a diagnosis of ACT. Typical imaging findings of pediatric ACT consist of a large, well-defined suprarenal tumor containing calcifications with a thin capsule and central necrosis or hemorrhage. The pathologic classification of pediatric ACT is troublesome. Even an experienced pathologist can find it difficult to differentiate carcinoma from adenoma. Surgery is the single most important procedure in the successful treatment of ACT. The role of chemotherapy in the management of childhood ACT has not been established although occasional tumors are responsive to mitotane or cisplatin-containing regimens. Because of the heterogeneity and rarity of the disease, prognostic factors have been difficult to establish in pediatric ACT. Patients with incomplete tumor resection or with metastatic disease at diagnosis have a dismal prognosis. In patients with localized and completely resected tumors, the size of the tumor has predictive value. Patients with large tumors have a much higher relapse rate than those with small tumors. PMID- 11004726 TI - The expression of the ACTH receptor. AB - Adrenal glucocorticoid secretion is regulated by adrenocorticotropic hormone (ACTH) acting through a specific cell membrane receptor (ACTH-R). The ACTH-R is a member of the G protein superfamily-coupled receptors and belongs to the subfamily of melanocortin receptors. The ACTH-R is mainly expressed in the adrenocortical cells showing a restricted tissue specificity, although ACTH is recognized by the other four melanocortin receptors. The cloning of the ACTH-R was followed by the study of this gene in human diseases such as familial glucocorticoid deficiency (FGD) and adrenocortical tumors. FGD is a rare autosomal recessive disease characterized by glucocorticoid deficiency, elevated plasma ACTH levels and preserved renin/aldosterone secretion. This disorder has been ascribed to an impaired adrenal responsiveness to ACTH due to a defective ACTH-R, a defect in intracellular signal transduction or an abnormality in adrenal cortical development. Mutations of the ACTH-R have been described in patients with FGD in segregation with the disease. The functional characterization of these mutations has been prevented by difficulties in expressing human ACTH-R in cells that lack endogenous melanocortin receptor activity. To overcome these difficulties we used Y6 cells, a mutant variant of the Y1 cell line, which possesses a non-expressed ACTH-R gene allowing the functional study without any background activity. Our results demonstrated that the several mutations of the ACTH-R found in FGD result in an impaired cAMP response or loss of sensitivity to ACTH stimulation. An ACTH-binding study showed an impairment of ligand binding with loss of the high affinity site in most of the mutations studied. PMID- 11004725 TI - Biosynthesis and metabolism of steroid hormones by human adrenal carcinomas. AB - Over a 15-year period, our university-based laboratory obtained 125 adrenal tumors, of which 15 (12%) were adrenal cortical carcinomas. Of these, 6 (40% of the carcinomas) occurred in patients with clear clinical manifestations of steroid hormone excess. Adrenal cortical carcinoma cells derived from the surgically resected tumors in 4 of these patients were isolated and established in primary culture. Radiotracer steroid interconversion studies were carried out with these cultures and also on mitochondria isolated from homogenized tissues. Large tumors had the lowest steroidogenic activities per weight, whereas small tumors had more moderately depressed enzyme activities relative to cells from normal glands. In incubations with pregnenolone as substrate, 1 mM metyrapone blocked the synthesis of corticosterone and cortisol and also the formation of aldosterone. Metyrapone inhibition was associated with a concomitant increase in the formation of androgens (androstenedione and testosterone) from pregnenolone. Administration of metyrapone in vivo before surgery in one patient resulted in a similar increase in plasma androstenedione, though plasma testosterone levels were not significantly affected. In cultures of two of four tumors examined, dibutyryl cAMP stimulated 11ss-hydroxylase activity modestly; ACTH also had a significant stimulatory effect in one of these tumors. Unlike results obtained with normal or adenomatous adrenal cortical tissues, mitochondria from carcinomatous cells showed a lack of support of either cholesterol side-chain cleavage enzyme complex or steroid 11ss-hydroxylase activity by Krebs cycle intermediates (10 mM isocitrate, succinate or malate). This finding is consistent with the concept that these carcinomas may tend to function predominantly in an anaerobic manner, rather than through the oxidation of Krebs cycle intermediates. PMID- 11004727 TI - Role of ACTH receptor in adrenocortical tumor formation. AB - Adrenocorticotrophin (ACTH) is the major regulatory hormone of steroid synthesis and secretion by adrenocortical cells. The actions of ACTH are mediated by its specific membrane receptor (ACTH-R). The human ACTH-R gene was recently cloned, allowing systematic determination of its sequence, expression and function in adrenal tumorigenesis. The presence of oncogenic mutations of the ACTH-R gene in adrenocortical tumors has been reported. Direct sequencing of the entire coding region of the ACTH-R gene of sporadic adrenocortical adenomas and carcinomas did not reveal constitutive activating mutations, indicating that this mechanism is not frequent in human adrenocortical tumorigenesis. Recent studies demonstrated allelic loss of the ACTH-R gene in a subset of sporadic adrenocortical tumors using a PstI polymorphism located in the promoter region of the ACTH-R gene. Loss of heterozygosity of the ACTH-R was analyzed in 20 informative patients with a variety of benign and malignant adrenocortical tumors. Three of them showed loss of heterozygosity of the ACTH-R gene. In addition, Northern blot experiments demonstrated reduced expression of ACTH-R mRNA in these three tumors with loss of heterozygosity, suggesting the functional significance of this finding at the transcriptional level. Deletion of the ACTH-R gene seems to be involved in a subset of human adrenocortical tumors, contributing to cellular dedifferentiation. PMID- 11004728 TI - Suicide attempts in the Hungarian adult population. Their relation with DIS/DSM III-R affective and anxiety disorders. AB - Prevalence of suicide attempts and their relationship with DIS anxiety and affective disorder diagnoses were investigated in a Hungarian adult community sample. Despite the high suicide mortality rate, the rate of suicide attempts was similar to that reported in other studies using similar methods. Suicide attempts occurred more frequently among women and previously married persons. Although the presence of any lifetime anxiety and/or affective disorder increased the rate of reported suicide attempts, the effect of co-morbidity, recurrence and chronicity might be considered significant predictors. The highest odds of an attempt were related to the diagnoses of dysthymic or bipolar disorders. Agitation was the only depressive symptom, which increased the odds of a suicide attempt. PMID- 11004729 TI - Deliberate self-poisoning presenting at a rural hospital in Northern Ireland 1976 1996: relationship to prescribing. AB - PURPOSE: This study reports on a project to monitor deliberate self-poisoning in a rural area of Northern Ireland over a 20-year period. Comparison is made with reports from large urban centres. In addition, a local prescribing database allows assessment of any association between psychotropic drug prescription and use for deliberate self-poisoning. MATERIALS AND METHODS: Frequency of self poisoning, demographic details and drugs used were recorded for all episodes of deliberate self-poisoning occurring at Craigavon Area Hospital for the years 1976, 1986, 1991 and 1996. It was possible to compare prescriptions of psychotropic drugs with their use for deliberate self-poisoning between the years 1991 and 1996 in the region served by the hospital, using the Defined Daily Dose (DDD) system. RESULTS: In this rural area the pattern of deliberate self poisoning has changed, as in urban centres, with a rise in frequency and the male/female ratio approaching unity. The pattern of drug use has altered, with paracetamol overtaking benzodiazepines as the most commonly used agent. More recently, antidepressants have become the second most frequently used drug class for this purpose. Psychotropic medications used for self-poisoning altered in proportion to their prescription between the years 1991 and 1996. CONCLUSIONS: In the face of a continuing rise in deliberate self-poisoning, which is effecting both urban and rural areas, care should be taken to prescribe the least toxic agent available as this is associated with likely frequency of self-poisoning for most classes of psychotropic drug. PMID- 11004730 TI - Prevalence of subthreshold forms of psychiatric disorders in persons making suicide attempts in Hungary. AB - BACKGROUND: Suicide and suicide attempts have been associated to psychiatric illnesses; however, little is known about the role in suicide risk of those symptoms that do not meet the full criteria for a DSM-IV disorder. The aim of this study was to examine the prevalence of subthreshold psychiatric disorders among suicide attempters in Hungary. METHODS: Using a modified structured interview (Mini International Neuropsychiatric Interview) determining 16 Axis I psychiatric diagnoses and their subthreshold forms defined by the DSM-IV and a semistructured interview collecting background information, the authors examined 140 consecutive suicide attempters, aged 18-65 years. RESULTS: Eighty-three-point six percent of the attempters had one or more current threshold diagnoses on Axis I and in addition more than three-quarters (78.6%) of the subjects had at least one subthreshold diagnosis. Six-point-four percent of the subjects (N = 9) had neither subthreshold nor threshold diagnoses at the time of their suicide attempts. Ten percent of the subjects (N = 14), not meeting the full criteria for any DSM-IV diagnoses, had at least one subthreshold diagnosis. In 68.6% of the subjects (N = 96), both subthreshold and threshold disorders were diagnosed at the time of their suicide attempts. The number of subthreshold and threshold diagnoses were positively and significantly related (chi2 = 5.12, df = 1, P < 0.05). Sixty-three-point-six percent of the individuals received two or more current threshold diagnoses on Axis I and 44.3% of the individuals (N = 62) had two or more subthreshold diagnoses at the time of their suicide attempts. LIMITATIONS: The subthreshold definitions in this study included only those forms of the disorders which required the same duration as the criteria DSM-IV disorder with fewer symptoms. Conclusions - Suicide attempts showed a very high prevalence of subthreshold disorders besides psychiatric disorders meeting the full criteria required according to the DSM-IV. Subthreshold forms of mental disorders need to be taken into account in suicide prevention. PMID- 11004731 TI - Evolution of the management of depression in Spain from the psychiatrist's perspective. A comparative analysis: 1997 vs 1982. AB - The main problem of depression is not only the high prevalence of the disorder but also its serious consequences on the patient's quality of life and the associated social costs in terms of health care resource utilization and productivity losses. In recent years, there has been a considerable improvement in the knowledge of depression from the pathogenic, clinical and therapeutic perspectives. The present study analyzes whether such advances are reflected in a positive evolution of the treatment of depression in Spain. To this effect we have contrasted the results of two socio-sanitary studies published in this country: the White Book editions of 1982 and 1997 (WB82 and WB97, respectively). From the methodological perspective, the physician selection criteria employed were very uniform (structured questionnaires delivered to 128 (WB82) and 300 (WB97) randomly selected psychiatrists). The origin of patients consulting for specialized care has varied over this 15-year period. In effect, WB82 patients were essentially referred by friends (87.5%) and from the primary care setting (44.5%), whereas in the WB97 study referral from primary care predominated (50.1%), followed by the patient's personal decision (24.8%). In turn, 40.7% and 51.7% of the psychiatrists in WB97 respectively considered the diagnostic and therapeutic means available in primary care to be insufficient. The priorities for improving patient quality of life, as reflected by both editions of the study, were the training of primary care physicians and the adequate provision of means in the mental health care centers. On the other hand, fewer problems for establishing a correct diagnosis were referred in the 1997 edition of the study (28.7%) than in 1982 (48.4%). In this sense, the main problem reported in WB82 was the lack of specialized training, whereas the masking of depression by some other disease process or symptoms was the main problem in WB97 (67.6% vs 21.1% according to WB82). The main symptoms upon which the diagnosis of depression are based do not seem to have evolved much in the past 15 years. The most frequently cited manifestations were a worsening of mood, loss of interest and leisure capacity, sleep alterations and diminished vitality. A comparative analysis of the therapeutic resources used was not possible, for prior to 1982 the only drugs available to physicians were the classical tricyclic agents and some MAO inhibitors; the selective serotonin reuptake inhibitors (SSRIs) - possibly the greatest advance in the treatment of depression in these 15 years - had not yet been introduced. Nevertheless, it should be pointed out that 98% of the psychiatrists consulted in WB97 considered pharmacologic treatment to be the most widely adopted form of management once depression has been diagnosed. PMID- 11004732 TI - P300 event-related brain potential and personality in depression. AB - P300 is an event-related brain potential (ERP) particularly interesting to the study of cognitive processes in normal subjects and in psychopathology. P300 has been applied in depression with controversial results. A major source for these controversial results could result from the diversity of depressed patients included in the different studies. Supporting this assumption, impulsivity, blunted affect, suicidal behavior and psychotic features significantly influence P300 amplitude. However, no data are available on the possible influences of the personality of depressed patients on P300. Since personality is related to P300 in normal subjects, the aim of the present study is to investigate the relationship between ERPs (P200, N200, and P300) and the Temperament and Character Inventory (TCI) in 54 depressed patients. The main results of the study concern the absence of major correlations between personality dimensions as assessed by the TCI and ERP parameters among depressed patients. Only weak partial positive correlations relate N200 latency with harm avoidance, and P300 amplitude (Pz) with the self-directedness dimension. N200 amplitude is also negatively correlated to persistence. However, the preliminary nature of the presented results with respect to the weak statistical significance should be underlined. PMID- 11004733 TI - Economic impact of using mirtazapine compared to amitriptyline and fluoxetine in the treatment of moderate and severe depression in the UK. AB - This study modelled the economic impact of mirtazapine, compared to amitriptyline and fluoxetine, in the management of moderate and severe depression in the UK, as well as the costs related to discontinuation of antidepressant treatment. Decision models of the management of moderate and severe depression were developed from clinical trial data, resource use obtained from interviews with general practitioners and psychiatrists, and published literature, and were used to estimate the expected direct National Health Service (NHS) costs of managing a patient with moderate or severe depression. The expected cost of healthcare resource use attributable to managing a patient suffering from moderate or severe depression who discontinues antidepressant treatment, irrespective of the initial treatment, was estimated to be pounds sterling 206 (range pounds sterling 50 to pounds sterling 504) over five months. Using mirtazapine instead of amitriptyline for seven months increases the proportion of successfully treated patients by 21% (from 19.2 to 23.2%) and reduces the expected direct NHS cost by pounds sterling 35 per patient (from pounds sterling 448 to pounds sterling 413). Using mirtazapine instead of fluoxetine for six months increases the proportion of successfully treated patients by 22% (from 15.6 to 19.1%), albeit for an additional cost to the NHS of pounds sterling 27 per patient (from pounds sterling 394 to pounds sterling 420). In conclusion, this study suggests that mirtazapine is a cost-effective antidepressant compared to amitriptyline and fluoxetine in the management of moderate and severe depression in the UK. PMID- 11004734 TI - Beneficial effect of the addition of the 5-HT 2A/2C and alpha2 antagonist mianserin to ongoing haloperidol treatment in drug-resistant chronically hospitalized schizophrenic patients. AB - Atypical neuroleptics that block serotonin 2A (5-HT 2A) and dopamine 2 (D2) receptors have been shown to possess efficacious antipsychotic activity. We assessed the efficacy of the addition of the 5-HT 2A/2C and alpha2 antagonist mianserin to ongoing haloperidol treatment in chronically hospitalized (> 10 years) drug-resistant schizophrenic patients (N = 12). The patients were assessed at baseline and every three months for one year with the Brief Psychiatric Rating Scale and the Clinical Global Impression. Results showed a significant (but < 10%) improvement in the core symptoms of schizophrenia; however, only the reduction (by 43%) in anxiety was clinically relevant (P < 0.0001). The beneficial effect of mianserin may be related to the combined blockade of 5-HT 2A and histamine (H1) receptors. PMID- 11004735 TI - Are washout periods useful in antidepressant trials? PMID- 11004736 TI - Protection from extinction in human fear conditioning. AB - Two experiments examined the ability of an added stimulus to interfere with extinction of a target excitatory fear stimulus (a predictor of shock) in human autonomic conditioning. Both experiments demonstrated disruption of extinction when the added stimulus was inhibitory (a predictor of no shock, or safety signal). Subjects showed a return of fear when the target stimulus was tested alone, on both self-reported shock expectancy and skin conductance measures. The second experiment also demonstrated disruption of extinction when the added stimulus was excitatory. This results suggests that protection from extinction may occur even when the added stimulus is not inhibitory. Additional factors that may contribute to protection from extinction include context-specificity, occasion-setting and external inhibition. The results highlight the role that concurrent stimuli play in extinction, and emphasise the need to keep concurrent stimuli as similar as possible to the desired transfer context in practical applications of extinction such as exposure therapy for anxiety. PMID- 11004737 TI - Does the social concerns component of the anxiety sensitivity index belong to the domain of anxiety sensitivity or the domain of negative evaluation sensitivity? AB - The present study utilized an exploratory factor-analytic approach (i.e. principal-components analysis; PCA) to investigate whether the Social Concerns component of the Anxiety Sensitivity Index (ASI [Peterson, R. A., & Reiss, S. (1992). Anxiety Sensitivity Index manual (2nd ed.). Worthington, OH: International Diagnostic Systems.]) is best conceptualized as belonging to the domain of anxiety sensitivity (AS) and/or the domain of negative evaluation sensitivity (NES). A sample of university students (N = 216) was administered measures of both NES (i.e. Brief Fear of Negative Evaluation scale; Leary, 1983) and AS (i.e. ASI). Participants' responses to the items comprising these measures were subjected to a PCA with oblique rotation. Factors representing the NES construct and the three lower-order AS constructs (i.e. AS Physical, Psychological and Social Concerns) were obtained. Subscales derived from these four factors were positively and significantly correlated with one another and loaded on a single higher-order factor labeled Threat Sensitivity. Thus, the present findings suggest that the AS Social Concerns factor is distinct from NES and the other lower-order components of AS. However, correlational analyses and higher-order PCA indicated that the AS Social Concerns factor taps a blend of AS and NES as well as something unique and distinct from both global AS and NES. PMID- 11004738 TI - Challenging the omnipotence of voices: group cognitive behavior therapy for voices. AB - The present study examines the impact of group based cognitive behavior therapy (CBT) for drug resistant auditory hallucinations, or voices. In particular it assesses treatment effect on beliefs in a voice's omnipotence and control. Twenty two participants entered one of five 8-session CBT groups. Measures of omnipotence, control, process measures, and symptoms of anxiety and depression were completed at assessment, and first and last group sessions. The groups achieved a significant reduction in conviction in beliefs about omnipotence (df 2, P = 0.002) and control (df 2, P = 0.001). There were no affective changes. Certain participants showed important spontaneous changes in behavior. Process measures suggested that participants valued the groups and benefited from them. These results are promising and the treatment may prove a useful addition to existing psychological interventions. PMID- 11004739 TI - Excessive breathlessness through emotional imagery in asthma. AB - Breathlessness and negative emotions during asthma attacks interact in complex patterns. This study tested the influence of emotional imagery on breathlessness during voluntary breath holding. Adolescents with and without asthma (n = 36 + 36) were assigned to positive imagery, negative imagery, or no imagery. There were four trials with close to thresholds for breath holding combined with imagery. Breathlessness and quality of imagery were measured by the end of breath holding. Additional measures were lung function and anxiety. The results showed that positive and negative imagery were only influencing breathlessness in participants with asthma. Although threshold duration for the groups were not significantly different, participants with asthma reported more breathlessness. The intensity of imagery enhanced breathlessness but diminished the accuracy of symptom perception. Positive imagery diminished breathlessness in participants with asthma, but also the difference in breathlessness between 75% and 95% of threshold duration. Breathlessness did not correlate with lung function, anxiety or other variables. It was concluded that emotional imagery during asthma attacks distracts from accurate introspection or enhances breathlessness, irrespective of anxiety. PMID- 11004740 TI - Posttraumatic stress disorder in breast cancer patients following autologous bone marrow transplantation or conventional cancer treatments. AB - We assessed 17 women who had undergone autologous bone marrow transplants (BMT) for their breast cancer and 20 other women who had been treated for breast cancer (but not with BMT) by structured clinical interviews examining each stage of the breast cancer experience (e.g. initial diagnosis, initial treatment, recurrence of cancer (if applicable) and BMT (if applicable)) and at follow-up points; 3, 6 and 12 months (if applicable) posttreatment. The two groups did not differ on incidence of posttraumatic stress disorder (PTSD), major depressive disorder (MDD) or generalized anxiety disorder at any stage. We found a high rate of PTSD over the cancer experience, 35% for the combined sample, with cancer diagnosis being the most likely point for developing PTSD, 27% for the combined sample. None of the 17 women who had undergone BMT developed PTSD as a result of the treatment. PMID- 11004741 TI - The relationship between inflated personal responsibility and exaggerated danger expectancies in obsessive-compulsive concerns. AB - The association between responsibility for a negative outcome, perceived severity of the outcome and perceived likelihood of the outcome was examined in a sample of 70 undergraduate students. Participants were asked to rate the likelihood and severity of 10 negative outcomes, five related to contamination and five related to checking. Thirty-eight participants completed a version of the questionnaire that presented the subject as responsible for the action that may lead to a negative outcome ('personally responsible' group). The remaining 32 completed a version of the questionnaire that presented someone else performing the actions that may lead to a negative outcome ('other responsible' group). Significant differences emerged between the personally responsible and other responsible groups for severity of outcome ratings but not for likelihood of outcome ratings. Specifically, for both washing and checking concerns, participants in the personally responsible group rated the severity of the potential negative outcome as greater than did those in the other responsible group. The results support the claimed general tendency for individuals to regard an outcome as more aversive if they are personally responsible for that outcome, rather than someone else being responsible. The results suggest that, in general, increasing perceptions of personal responsibility will increase cost or severity estimates in subjective danger calculations, and that responsibility may influence OCD phenomena in this way. Finally, the results suggest that attempts to manipulate responsibility in the laboratory may be confounded by necessarily impacting on cost estimates, and therefore on danger expectancies. PMID- 11004742 TI - The cardiac anxiety questionnaire: development and preliminary validity. AB - Heart-focused anxiety (HFA) is the fear of cardiac-related stimuli and sensations because of their perceived negative consequences. Although HFA is common to a wide variety of persons who experience chest pain and distress, it often is unrecognized and misdiagnosed, particularly in cardiology and emergency room patients without and with heart disease. To address these concerns, this article reports on the development and preliminary psychometric evaluation of the Cardiac Anxiety Questionnaire (CAQ) designed to measure HFA. In Study 1, 188 cardiology patients completed the CAQ. Item and factor analyses indicated a three-factor solution pertaining to heart-related fear, avoidance, and attention. Reliability analysis of the 18-item CAQ revealed good internal consistency of the total and subscale scores. In Study 2, 42 patients completed the CAQ and several other anxiety-related questionnaires to assess its convergent and divergent properties. Although preliminary validity results are promising, further psychometric study is necessary to cross-validate the CAQ, examine its test-retest reliability, and confirm the stability of the factor structure. Taken together, the CAQ appears to assess HFA, and may therefore be a useful instrument for identifying patients with elevated HFA without and with heart disease. PMID- 11004743 TI - [Study of the brain involuntary syntactic processing by positron emission tomography]. PMID- 11004744 TI - [Effect of glycine on EEG parameters and the sensorimotor activity in normal and alcoholic subjects]. PMID- 11004745 TI - [Amplitude modulation of EEG in humans]. PMID- 11004746 TI - [Right and left hemispheres in the emotional evaluation of visual stimuli]. PMID- 11004747 TI - [Dependence of the threshold of sinusoidal grating displacement on its spatial frequency characteristics]. PMID- 11004748 TI - [Study of latent periods of visual object differentiation using various significant characteristics]. PMID- 11004749 TI - [Regulation of blood circulation in hypertonic crises using mathematical analysis of the heart rhythm]. PMID- 11004750 TI - [Analysis of the heart rhythm variability in negative functional states during sessions of psychological relaxation]. PMID- 11004751 TI - [Effect of extralow frequency variations of atmospheric pressure on voluntary attention parameters]. PMID- 11004752 TI - [Electrical activity of the triceps sural muscle under static load in patients with chronic peripheral circulatory insufficiency]. PMID- 11004753 TI - [Effect of various regimes of interval hypoxic training on cardiorespiratory and hematologic functions]. PMID- 11004754 TI - [The role of thoracic and abdominal components of the respiratory system during hyperventilation combined with chemoreceptor stimulation of various intensity]. PMID- 11004756 TI - [Circadian rhythms of salivary cortisol level during long-term space flight]. PMID- 11004755 TI - [Physical working capacity and its determining factors during 120 day antiorthostatic hypokinesia]. PMID- 11004757 TI - [Microrheological properties of various erythrocyte populations in patients with elevated blood pressure and in physically active subjects]. PMID- 11004758 TI - [Constitutional characteristics of response of hemostasis to the physical load]. PMID- 11004759 TI - [Does the respiratory system limit the human aerobic work capacity?]. PMID- 11004760 TI - [Does the 48-hour biological rhythm exist?]. PMID- 11004761 TI - [Reaction time during perception of acoustic models of sound approaching and withdrawing under backward masking conditions]. PMID- 11004762 TI - [Interhemisphere interactions during bimanual backward masking]. PMID- 11004763 TI - [Sex difference in the electrodermal activity under various wakefulness conditions]. PMID- 11004764 TI - [Constitutional aspects of psychoemotional stresses in adolescents]. PMID- 11004765 TI - [Living donor liver transplantation in Kyushu University]. AB - We performed living donor liver transplantation (LDLT) for 40 patients at Kyushu University Hospital, Fukuoka Japan during the period from October 1996 to April 2000. The patients consisted of 32 adults and 8 children with a mean age of 35.8 years (range: 1 year and 10 months to 65 years old). The underlying liver diseases of the 40 patients included the fulminant hepatic failure (n = 14), biliary atresia (n = 7), liver cirrhosis (HCV) (n = 6), primary biliary cirrhosis (n = 5), primary sclerosing cholangitis (n = 2), familiar amyloidotic polyneuropathy (n = 2), Alagille syndrome (n = 1), glycogen storage disease (n = 1), huge hepatic hemangiomas (n = 1), and Wilson's disease (n = 1). All liver grafts were obtained from each patient's family members except for one domino transplant donor's case, comprised of 13 parents, 13 sons and daughters, 11 brothers and sisters, and 3 wives. The donors are presently all doing well. The patient survival rate is presently 92.5%. PMID- 11004766 TI - [Lung cancer risk and genetic polymorphism at the glutathione S-transferase P1 locus in male Japanese]. AB - The glutathione S-transferases (GSTs) are widely expressed in mammalian tissues and involved in Phase II detoxification reactions. The GSTs form a supergene family consisting of four distinct families, named alpha (GSTA), mu (GSTM) theta (GSTT) and pi (GSTP). Several of the GST genes are polymorphic in humans and are currently being investigated as possible cancer-risk modifiers. Among the GST genes, we examined GSTP1 polymorphism in exon 5 among male lung cancer patients (n = 86, male Japanese) and male healthy controls (n = 80, male Japanese) by restriction fragment length polymorphism-polymerase chain reaction method. The cancer patients showed frequency of the GSTP1 mutated genotype (individuals having at least one mutant allele, 29.1%) very similar to that of the control subjects (28.8%). After adjusting for smoking status, no association was found between the GSTP1 mutated genotype and lung cancer risk (odds ratio: 0.95; 95% confidence interval: 0.48-1.90). When study subjects were divided into two subgroups based on smoking status, the GSTP1 mutated genotype was not associated with an increased risk of lung cancer among smokers and non-smokers. These results suggest that GSTP1 polymorphism in exon 5 alone may not increase the risk of lung cancer in male Japanese. PMID- 11004767 TI - Life's many pieces: the identity of the group psychotherapist. AB - The American Group Psychotherapy Association (AGPA) and its members are experiencing a trauma in response to the changes in health care imposed by the managed care industry. Although the economic threat is frightening, the threat to professionals' and AGPA's identities is even more damaging. The increasingly rapid, society-wide rate of change and factors related to the aging of AGPA's membership complicate its ability to cope with the trauma. The author recounts personal trauma and professional experiences in treating traumatized individuals, and proposes that when certain needs are met, growth can occur as part of a healing process: getting through the trauma, finding safety, self-soothing, and achieving mastery. AGPA resources for being safe, self-soothing, and achieving mastery are described. AGPA has been successful in reaching out via new accomplishments in regard to visibility, although the understandable impulse to withdraw may be impeding efforts to diversify the membership body. The author urges fighting against the pull to withdraw and expresses hope that the AGPA will cope well with the trauma by using its significant resources and will ultimately experience a beneficial change in its identity. PMID- 11004768 TI - A sociohistorical view of group psychotherapy in the United States: the ideology of individualism and self-liberation. AB - A deep strain of individualism permeates American culture, rooted in the political-economic ideology of capitalism. The ideal of a self-governing individual is promoted, independent from social, historical, and cultural forces, whose thoughts and emotions are located within the construct of a masterful self, ready to be filled and expanded. This article traces the influence of self liberatory ideology in American group psychotherapy through the religious revivalist and Mental Hygiene movements. The "progressivism" of pioneering group theorists is examined in terms of revisionist psychology and self-liberatory practice. Psychodrama and T-groups are demonstrated to be precursors to the encounter group movement in which the belief in self-liberation reached its zenith. Early group analytic approaches are seen to eschew transpersonal and group dynamic processes. PMID- 11004769 TI - The future of the group. AB - Group oppositional and destructive processes have been insufficiently understood and integrated into the fabric of group psychotherapy. To strengthen our field for the future, this area requires further elucidation. The author's concept of the Anti-Group is used as a basis for exploring antagonistic group processes in the clinical setting in parallel to group disruption and threat in the social domain. Further, it is suggested that the legacy of our training organizations is tied to problematic and institutional group processes that tend to be repeated rather than acknowledged and resolved. These gaps in our understanding limit the power of the group because creativity and growth come from the recognition and transformation of destructive impulses. PMID- 11004770 TI - Community meeting: review, update, and synthesis. AB - The community meeting is a frequent regular meeting in a psychiatric ward or institution that is attended by all staff and patients. It meets for the purpose of communication, ward management, or psychiatric treatment. The conditions of the current hospital psychiatric treatment environment are very different from the treatment environment where the therapeutic community concept was first established, particularly in terms of the shortened length of hospital stay and the emphasis on pharmacologic rather than psychotherapeutic methods of treatment. However, even with these changes, an admixture of psychopharmacologic treatment and therapeutic community principles has been advocated as an effective, realistic, and desirable treatment modality in today's hospital environment. A synthesis of an extensive literature review from both a theoretical and research perspective on the community meeting is presented. A schema has been developed listing elements for practice in today's hospital climate. PMID- 11004771 TI - An integrated model of group treatment for cancer patients. AB - Considerable evidence suggests that group interventions are a valuable resource for cancer patients, but few conceptual frameworks are available to guide decisions about which approaches might be most useful for which patients at what phases of illness. This article presents an integrative treatment model for group services. It describes different group interventions geared toward patients at different phases of illness to accommodate the shifting needs and concerns that evolve over the course of the disease. Recommendations are offered concerning further research and model development. PMID- 11004772 TI - Further comments on understanding group psychotherapy by Irvin Yalom. PMID- 11004773 TI - Growth through shame and humiliation. PMID- 11004774 TI - Feasibility of controlling Ixodes scapularis ticks (Acari: Ixodidae), the vector of Lyme disease, by parasitoid augmentation. AB - A theoretical analysis of the feasibility of controlling tick populations (Ixodidae) by the release of reared Ixodiphagus parasitoids in tick ecosystems yielded promising results. The analysis suggested that if reasonable progress could be made in mass-rearing the parasitoids, it would be possible to control the blacklegged tick, Ixodes scapularis (Say), the vector of Lyme disease, by this biological control procedure. Lyme disease has become the most important vector-borne disease in the United States. In a field-release experiment conducted in Africa by members of the International Center for Insect Physiology and Ecology, effective control of Amblyomma variegatum (F.) was obtained by the release of Ixodiphagus parasitoids in tick habitats. Encouraging theoretical results along with the encouraging results of a field-release experiment indicate the need for civil and political leaders in countries where ticks are a major problem to sponsor strong and well-coordinated research initiatives focused on the development of this new method of dealing with tick problems. PMID- 11004775 TI - Coadaptation of male aedeagal filaments and female spermathecal ducts of the old world phlebotomine sand flies (Diptera: Psychodidae). AB - The morphology of insect genitalia is often highly species-specific, and its variation has been suggested as an important impetus for evolution. Structural variation of the male genitalia and the female spermathecae in Phlebotomine sand flies is unique among the blood sucking Diptera. We describe the fine structures involved in mating for Phlebotomus papatasi (Scopoli). Relationships among the length of the spermathecal duct and aedeagal filament were studied in 26 species of Old World phelebotomine sand flies comprising 12 subgenera. In most taxa the aedeagal filaments were very long allowing direct insemination into the spermathecae, indicating that intrasexual competition among males occurs as sperm precedence. There was a positive correlation between the lengths of the spermathecal ducts and aedeagal filaments, indicating that an evolutionary "arms race" occurs between males and females over the control of fertilization. The pattern of genitalia variation in phlebotomine sand flies also indicated that differences in the lengths of the spermathecal ducts and aedeagal filaments are distributed unevenly among closely related species. Genital differences do not seem to occur in populations of the Phlebotomus argentipes Annandale & Brunetti complex, whereas differences showed strongly in closely related Phlebotomus papatasi and P. bergeroti Parrot, and in sympatric Phlebotomus martini Parrot and P. celiae Minter. PMID- 11004776 TI - Vector competence of Aedes aegypti, Culex sitiens, Culex annulirostris, and Culex quinquefasciatus (Diptera: Culicidae) for Barmah Forest virus. AB - Aedes aegypti (L.), Culex sitiens Weidemann, Culex annulirostris Skuse, and Culex quinquefasciatus Say mosquitoes colonized at the Queensland Institute of Medical Research, Brisbane Australia, were fed on blood containing Barmah Forest virus (BF). Only Cx. annulirostris was susceptible to infection, with a median cell culture infectious dose (CCID50) of 10(3.36) per mosquito. Ae. aegypti and Cx. quinquefasciatus were infected experimentally, but at rates of < 9%. Cx. sitiens did not become infected. Infection rates for Cx. annulirostris fed 10(3.5) CCID50 of virus per mosquito, varied from 9 to 50% between 2 and 13 d after infection. Virus transmission to suckling mice by Cx. annulirostris occurred from 2 d after infection. Transmission of BF virus by Cx. annulirostris was 10% at 2 d after infection and did not exceed 8% thereafter. Although Cx. annulirostris may be infected and is able to transmit BF virus to suckling mice, it is nonetheless a relatively inefficient vector of the virus. PMID- 11004777 TI - Early warning of malaria epidemics in African highlands using Anopheles (Diptera: Culicidae) indoor resting density. AB - Several highland regions of Africa recently have suffered malaria epidemics. Because malaria transmission is unstable and the population has little or no immunity, these highlands are prone to explosive outbreaks when densities of Anopheles exceed critical levels and conditions favor transmission. If an incipient epidemic can be detected early enough, control efforts may reduce morbidity, mortality, and transmission. Here we present three methods (direct, minimum sample size, and sequential sampling approaches) that could be used to determine whether the household indoor resting density of Anopheles gambiae s.I. has exceeded critical levels associated with epidemic transmission. Data on Anopheles density before, during, and after a malaria epidemic (December 1997 July 1998) in the highlands of southwestern Uganda were evaluated to demonstrate the application of these three approaches. During this epidemic, a density of 0.25 Anopheles mosquitoes per house was associated with epidemic transmission, whereas 0.05 mosquitoes per house was chosen as a normal level expected during nonepidemic months. The direct approach to calculating mean Anopheles density with an allowable error of 20-50% of the mean would require the sampling of 102 16 houses, respectively. In contrast, with only seven houses, the minimum sample size approach could be used to determine whether Anopheles density had exceeded the critical level. This method, however, would result in an overestimation of the risk of an epidemic at low Anopheles density. Finally, a sequential sampling plan could require as many as 50 houses to conclude that risk of an epidemic existed, but this disadvantage is offset by the ability to preset the probabilities of concluding that risk of an epidemic exists at both the critical and normal Anopheles densities. Our study illustrated that it is feasible, and probably expedient, to include monitoring of Anopheles density in highland malaria epidemic early warning systems. PMID- 11004778 TI - Seasonal abundance and survivorship of Culicoides sonorensis (Diptera: Ceratopogonidae) at a southern California dairy, with reference to potential bluetongue virus transmission and persistence. AB - Seasonal abundance and survivorship of Culicoides sonorensis Wirth & Jones were examined at a dairy in southern California from January 1995 to December 1997. Insects were collected one to two times per week using five CDC-type suction traps (without light) baited with CO2 at a constant release rate of 1,000 ml/min. Female and male abundance was greatest during late summer and early fall and was directly correlated with mean monthly air temperature. Parity of females was lowest during late summer and early fall. The gonotrophic cycle was estimated to require 3-4 during hot summer months and up to 14 d during cool winter months. Estimated extrinsic incubation of bluetongue virus (BLU) was 9-10 d during August and September. The estimated daily survival ranged from < 60% in the summer to > 95% in the winter, resulting in an expectation of life of only 2-3 d in summer and > 10 d in winter. The probability of females surviving the extrinsic incubation period for BLU virus, and the expectation of infective life were both lowest during late summer and early fall. During 1997, midge abundance during late summer was not high enough to overcome very low survivorship, and the absolute number of females expected to survive the extrinsic incubation period was relatively low. However, in 1995 and 1996, very high midge abundance compensated for low survivorship during summer and the number of females expected to survive the extrinsic incubation period was relatively high. Although abundance was generally very low during the cool winter and spring, host-seeking females were captured throughout the year, and their winter survival was high. Overwintering of BLU virus by continued transmission of the virus by active midges appears possible. PMID- 11004779 TI - Effects of temperature and pH on hexokinase from the flight muscles of Dipetalogaster maximus (Hemiptera: Reduviidae). AB - Effects of temperature and pH on the catalytic properties of hexokinase (HK, EC 2.7.1.1) from the flight muscles of Dipetalogaster maximus (Uhler) were studied. The enzyme showed a hyperbolic behavior with its two substrates (glucose and ATP). There was no inhibition by glucose. Apparent Km and Vmax increased as pH increased from 7.0 to 8.5. Catalytic efficiency was lowest at pH 7.0. Km, Vmax, and catalytic efficiency were higher at 37 degrees C than at 30 and 20 degrees C. There was marked inhibition by ATP, which was dependent on pH and temperature. Km values for ATP were reduced and catalytic efficiency increased as pH increased. Lowest Vmax was observed at pH 7.0. At this pH there was 87.3% inhibition by ATP, whereas it was only 5.7% at pH 8.5 (at 30 degrees C). Km, Vmax, and catalytic efficiency were higher at 37 degrees C than at 30 and 20 degrees C. The strong inhibition by ATP detected at 20 degrees C (pH 7.6) almost disappeared at 37 degrees C. Therefore, temperature could regulate hexokinase activity by modulating the inhibition produced by ATP. Glucose utilization and ATP production would be promoted when temperature rises from 30 to 37 degrees C. Because insect thoracic muscles increase their temperature over 30 degrees C during flight, this phenomenon elucidates a mechanism enhancing energy supply for muscle activity. PMID- 11004780 TI - Density and distribution of Dirofilaria immitis (Nematoda: Filarioidea) third stage larvae in Aedes sollicitans and Aedes taeniorhynchus (Diptera: Culicidae). AB - Numbers and the distribution of third-stage larvae (L3) were investigated in Aedes sollicitans (Walker) and Aedes taeniorhynchus (Wiedemann) female mosquitoes fed Dirofilaria immitis (Leidy) infectious-blood with densities of microfilariae (mf) ranging from approximately 7,100-43,400 mf/ml. At each microfilarial density, a maximum of 63-66 infective larvae were recovered from an Ae. sollicitans alive on day 15 after infection. In comparison with Ae. taeniorhynchus, Ae. sollicitans averaged greater numbers of L3 and from 1.4 to 2.4 times more L3 in the head and labium per infected female. The trend was for greater numbers of L3 to be found in the labium than in the head of Ae. sollicitans, but there were no significant differences between numbers of L3 recovered from these sites in differentially infected females. However, numbers of L3 recovered from the head versus the labium of differentially infected Ae. taeniorhynchus varied significantly with the infectious blood microfilarial density. At the two lowest and highest microfilarial densities, greater numbers of L3 were recovered from the head and labium, respectively. Variations among species in the female body size, blood meal size, and retention of L3 may be factors responsible for differences observed between the total numbers and percentage distribution of D. immitis L3 recovered from the simultaneous blood fed Ae. sollicitans and Ae. taeniorhynchus. PMID- 11004781 TI - Effect of eastern equine encephalomyelitis virus on the survival of Aedes albopictus, Anopheles quadrimaculatus, and Coquillettidia perturbans (Diptera: Culicidae). AB - The effect of eastern equine encephalomyelitis (EEE) virus on the survivorship of Aedes albopictus (Skuse), Anopheles quadrimaculatus Say, and Coquillettidia perturbans (Walker) was determined experimentally. Female mosquitoes were allowed to feed on EEE viremic chicks, and survival rates were compared for infected and uninfected mosquitoes. Additionally, the survival of female Cq. perturbans and An. quadrimaculatus intrathoracically (i.t.) inoculated with EEE was compared with controls receiving diluent inoculations. Infection with EEE significantly reduced survival in Cq. perturbans compared with uninfected individuals in per os infection experiments. I.t. infections of Cq. perturbans did not reduce survival when compared with diluent inoculated groups. In contrast, infection with EEE did not affect the survival of Ae. albopictus after per os infection or An. quadrimaculatus after either i.t. or per os infections. PMID- 11004782 TI - Characterization of a new spotted fever group rickettsia detected in Ixodes ricinus (Acari: Ixodidae) collected in Slovakia. AB - Two previously undescribed rickettsiae were detected in Ixodes ricinus Ricketts by polymerase chain reaction. Ixodes ricinus Slovakia (IRS) 3 and IRS4 were identified in ticks collected in northeastern and southwestern Slovakia, respectively. Sequences of the 16S rRNA citrate synthase (gltA) and outer membrane protein rOmpA (ompA) encoding genes of both strains were nearly identical but were distinct from those of all other known rickettsiae. Phylogenetic relationships inferred from the comparison of these sequences with those of other members of the genus Rickettsia indicate that IRS3 and IRS4 constitute a new rickettsial genotype and form a separate cluster among the spotted fever group rickettsiae. PMID- 11004783 TI - Life cycle of Ixodes (Ixodes) loricatus (Acari: Ixodidae) under laboratory conditions. AB - The life cycle of Ixodes (Ixodes) loricatus Neumann, reared in the laboratory, is described. Engorged females collected from opossums trapped in the states of Minas Gerais and Sao Paulo, Brazil, which were used to start the laboratory colonies, were designated as BMG and CSP, respectively. Larval and nymphal ticks from both colonies fed separately on Rattus norvergicus Berkenhout or Calomys callosus Rengger, whereas Didelphis marsupialis L and Didelphis albiventris Lund were used as hosts for BMG and CSP adults, respectively. Biological and developmental data obtained from ticks of both the BMG and CSP colonies that were reared separately for two consecutive generations were compared. The percentage of fed or molted ticks reared on C. callosus was higher than that recorded for ticks fed on R. norvergicus in the majority of the observations. Despite significant differences among several of the biological parameters, the pattern of the life cycles of the two tick colonies was similar. Results indicated that the mean life cycle duration of I. (I.) loricatus was approximately 7 mo from parental oviposition to the occurrence of F1 eggs, regardless of geographic origin or host species. PMID- 11004784 TI - Resistance to insecticides and effect of synergists on permethrin toxicity in Pediculus capitis (Anoplura: Pediculidae) from Buenos Aires. AB - Permethrin-resistant colonies of Pediculus capitis (De Geer) from Buenos Aires were used to establish a resistance profile and to examine resistance mechanisms. All permethrin-resistant head lice (resistance ratio from 52.8 to > 88.7) were also resistant to d-phenothrin (resistance ratio from 40.86 to > 48.39) and deltamethrin (resistance ratio from 16.24 to 38.06). No cross-resistance to carbaryl was found in any of the pyrethroid-resistant P. capitis tested. Otherwise, all resistant colonies showed low to high levels of resistance to beta cypermethrin. This pyrethroid had never been applied as a pediculicide in Argentina; however, the high level of resistance found in these permethrin resistant colonies (resistance ratio from 9.74 to 50.97) demonstrated that pyrethroid cross-resistance occurred to this novel insecticide. Treatment with piperonyl butoxide (PBO) or triphenylphosphate (TPP) significantly decreased the toxicity of permethrin in the four colonies tested. The esterase inhibitor TPP produced lower enhancement of toxicity than the multifunction oxidase inhibitor PBO in the colonies having the highest resistance levels. Results presented here concerning the cross-resistance profile and synergism by enzyme inhibitors in permethrin-resistant head lice demonstrated that enhanced metabolism was involved in the pyrethroid resistance. However, the substantial degree of resistance that remained after synergism suggested the presence of another resistance mechanism. Cross-resistance to pyrethroid and susceptibility to the carbamate carbaryl suggested a common action mechanism. PMID- 11004785 TI - Lipid utilization for ovarian development in an autogenous mosquito, Culex pipiens molestus (Diptera: Culicidae). AB - During the first ovarian cycle, autogenous female mosquitoes develop their ovaries in the absence of blood feeding. In autogenous Culex pipiens molestus (Forskal), complete yolk deposition was observed 2 d after emergence, even when no feeding was allowed (starved). Neutral lipids in Cx. p. molestus increased during the pupal stage, abruptly declined after emergence, and again increased on day 3. In contrast, neutral lipids decreased in anautogenous Anopheles stephensi (Liston) and Cx. p. pallens (Coquillett) and starved females died within 2-3 d after emergence. High ratios of two major neutral lipids, free fatty acid and triglyceride, were isolated by thin-layer chromatography (TLC) from the lipid contents of both Cx. p. molestus and An. stephensi fourth-instars and newly emerged females. Fatty acid analyses using gas liquid chromatography (GLC) and GLC-mass spectrometry (GLC-MS) showed higher proportions of unsaturated than saturated fatty acids in Cx. p. molestus at both stages and two major neutral lipids: free fatty acids and triglycerides. The percentage composition of linoleic acid (C18:2), which is a precursor of arachidonic acid, was higher in Cx. p. molestus than in An. stephensi. Our results indicated that elevated lipid content before emergence may play a role of inducing ovarian development in autogenous mosquitoes. PMID- 11004786 TI - Effects of density and larval competition on selected life history traits of Culex pipiens quinquefasciatus (Diptera: Culicidae). AB - The effects of larval densities of one to four individuals in standard Drosophila vials (diameter 25 by 95 mm) on the age at pupation, starved dry weight, and wing length of Culex pipiens quinquefasciatus Say were studied. This approach required relatively few larvae per replicate and included a control treatment, where individual larvae developed in the absence of competition. This design also tested for competitive interactions between male and female larvae. Mosquitoes pupated later, and emerged with lighter starved dry adult weight and shorter wings as larval density increased. The size of adult female mosquitoes, particularly their starved dry weight, was sensitive to larval density and also was influenced by the presence or absence of competition with another female larva. In contrast, the life history traits of males did not vary as a function of competition with female larva. Female larvae were also more likely to die in the highest density treatment. This design confirmed previous results and offered a potentially useful experimental approach to investigate the effects of density dependent competition among mosquito larvae. PMID- 11004787 TI - Definition of species in the Culex sitiens subgroup (Diptera: Culicidae) from Papua New Guinea and Australia. AB - Japanese encephalitis is a disease of major importance in Asia, which from 1995 has extended its range into the north of Australia. Culex annulirostris Skuse of the Culex sitiens subgroup of Culex was incriminated as the most likely mosquito vector. Because misidentifications previously have occurred within this subgroup, our paper defines electrophoretic methods to identify Culex specimens from Queensland and the Northern Territory in Australia and from Western Province in Papua New Guinea. In collections from 1996 to 1998 Culex sitiens Wiedemann, Cx. palpalis Taylor, and Cx. annulirostris comprised 93.1% of the total banded proboscis Culex mosquitoes collected; 6.9% remained unidentified to species by morphological or electrophoretic examination. The prevalence of Cx. palpalis was variable and appeared to be associated with below average rainfall during 1997. In Papua New Guinea, this species comprised up to 97.4% of banded proboscis Culex, with smaller numbers collected from the Torres Strait Islands and Cape York; none was collected from other Queensland localities. Cx. annulirostris was more prevalent during years of average or above average rainfall and was predominant in most localities, especially in the Torres Strait and Queensland. These data confirmed the problems in separating Cx. sitiens from Cx. palpalis and Cx. annulirostris using morphological characters. PMID- 11004788 TI - Simulium vittatum (Diptera: Simuliidae) and Lutzomyia longipalpis (Diptera: Psychodidae) salivary gland hyaluronidase activity. AB - Hyaluronidase activity in the salivary gland homogenates of Simulium vittatum (Zetterstedt) is described, and its optimal pH determined. Salivary activity was reduced significantly after a blood meal, indicating that it was secreted after blood feeding. Phlebotomus papatasi (Scopoli) also exhibited salivary hyaluronidase activity. These results indicate that hematophagous pool-feeding insects may secrete this enzyme to help the spread of salivary antihemostatic agents in the vicinity of the feeding lesion, and perhaps to increase the size of the feeding lesion itself. Additionally, this enzyme may affect local host immune reactions and promote arboviral transmission. PMID- 11004789 TI - Comparative study of the mouthparts of males and females of Lutzomyia migonei (Diptera: Psychodidae) by scanning electron microscopy. AB - The mouthparts of both males and females of Lutzomyia migonei Franca were compared by scanning electron microscopy (SEM) and light microscopy (LM), and their structures are herein described. Mandibles are present in both sexes but in males they are reduced and, in contrast to females, no teeth were observed. As in females, the laciniae in males are highly sclerotized, but the number of lateral teeth are reduced and retrorse teeth are absent. Only females possess teeth at the tip of the hypopharynx, whereas in males they are substituted by spicula. Except for the length from the junction with the hypopharynx to the tip, which is shorter in males than in females, the labrum is similar in both sexes in terms of form and sensilla. On the labrum of males sensilla could be detected similar to the apical sensilla of females which play a role in the identification of blood. PMID- 11004790 TI - Effects of local geographic barriers and latitude on population structure in Anopheles punctipennis (Diptera: Culicidae). AB - We sampled Anopheles punctipennis (Say) from 11 localities throughout Vermont to examine the effects of latitude and two local geographical boundaries, Lake Champlain and the Green Mountains, on the population genetic structure of this species. Thirty-five mitochondrial haplotypes were detected in 104 individuals using a variable region of the COI gene. When latitude was examined, we detected significant structure within localities and among localities within latitudinal regions. For geographic analysis, significant genetic structure was detected only within localities. Estimates of gene flow across geographic regions indicate that the Green Mountains, but not Lake Champlain, is a barrier to dispersal for this species. We found no correlation between genetic and geographic distances for An. punctipennis. PMID- 11004791 TI - Production of pheromones by artificially fed males of the tick Amblyomma maculatum (Acari: Ixodidae). AB - Aggregation-attachment pheromones are produced only by male ticks of the genus Amblyomma that have imbibed blood for at least 8 d from their mammalian hosts. This report demonstrates that production and release of aggregation-attachment pheromones by Amblyomma maculatum (Koch) males can be induced in vitro by using nonblood media, artificially introduced through capillary tubes, into the male ticks. The presence of these pheromones is demonstrated by using both biological observations on bovine hosts and in vitro experimentation with petri dishes. The attraction of unfed female ticks to the artificially fed males (using the petri dish method) was significantly higher (P < 0.001) than the attraction of unfed females to unfed males. Similarly, attraction and attachment were recorded on bovine hosts. Only 62.8 +/- 17.5% of the females released attached to the bovine host. Of these, 61.9 +/- 19.37% attached around artificially fed males that were placed on the bovine 24 h earlier. This percentage did not differ significantly from the 81.4 +/- 7.1% of the total that attached around males that fed naturally for 8 d. In comparison, only 33.3 +/- 21.9% of the total number of females placed on the host did so after being fed 1 d (control group). The possibility of using capillary feeding as a new tool for investigating the physiology and reproductive behavior of blood-sucking arthropods is discussed. PMID- 11004792 TI - Vector competence of Ixodes muris (Acari: Ixodidae) for Borrelia burgdorferi. AB - The vector competence of Ixodes muris (Bishopp & Smith) was determined for Borrelia burgdorferi, the etiologic agent of Lyme disease. Larval I. muris were fed on ICR outbred mice infected with the B-31 laboratory strain of B. burgdorferi. Replete larvae, at 5 d after feeding, were assayed for infection by culture in Barbour-Stoner-Kelly (BSK-H) media. Infection frequency in I. muris replete larvae was 66%. Resultant nymphs were fed on naive ICR outbred mice to determine the ability of I. muris to transmit infection. Infection frequency in fed nymphs declined to 38% and only 1/5 mice was positive for B. burgdorferi on ear biopsy culture. We demonstrated that I. muris is capable of acquiring and transmitting B. burgdorferi but is a relatively poor vector compared with I. scapularis (Say). PMID- 11004793 TI - Spectrofluorometers are not adequate for aging Aedes and Culex (Diptera: Culicidae) using pteridine fluorescence. AB - Attempts were made to use pteridine fluorescence levels as a tool for aging individual Aedes polynesiensis Marks and Culex quinquefasciatus Say mosquitoes. Fluorescent pigments were extracted and quantified with techniques and apparatus (i.e., a spectrofluorometer) that can be used in developing countries or in field laboratories, and that has already given consistent results in similar studies on other insect groups. However, for Ae. polynesiensis as well as Cx. quinquefasciatus, individual fluorescence measurements were mostly below the spectrofluorometer white noise level. With batches of Aedes or Culex of the same age, significant fluorescence levels were recorded but not in relation to their calendar ages. The low content of pteridines in mosquitoes suggests that standard spectrofluorometry is not sufficiently sensitive for such studies. PMID- 11004794 TI - Ticks (Acari: Ixodidae) associated with rural dogs in Uruara, eastern Amazon, Brazil. AB - A survey of 124 dogs for ticks was conducted from August to November 1998 in 68 small farms of Uruara municipality in Eastern Amazon-Brazil. We collected 55 ticks from 18 dogs (14.5%). Amblyomma oblongoguttatum Koch, Amblyomma ovale Koch, and Amblyomma cajennense (F.) were found on nine, six, and one dog, respectively. Fourteen small farms (20.5%) had at least one dog with ticks. PMID- 11004795 TI - A case of gynandromorphism in Amblyomma oblongoguttatum (Acari: Ixodidae). AB - During a survey conducted from August to November 1998 to ticks associated with dogs in the rural area of Uruara, in the state of Para, Eastern Amazon region of Brazil, we collected one gynandromorph specimen out of 18 Amblyomma oblongoguttatum Koch specimens collected from dogs. It is a perfect bipartite gynandromorph, reported in this species for the first time. PMID- 11004796 TI - Vector competence of Culex pipiens molestus (Diptera: Culicidae) from Taiwan for a sympatric strain of Japanese encephalitis virus. AB - Laboratory strains of Culex pipiens molestus Forskal and Culex tritaeniorhynchus Giles from northern Taiwan were compared for their susceptibility to the Sanhsia MQ1-2 (SH) strain of Japanese encephalitis (JE) virus isolated from Taiwan. After feeding on a sweetened blood-virus mixture, viral titers in Cx. p. molestus during the 14-d incubation period ranged from a minimum of 2.9 log10PFU (plaque forming units) per mosquito on day 3 after ingestion to a maximum of 4.65 log10PFU at day 8 and in Cx. tritaeniorhynchus from 2.6 on day 10-5.18 log10PFU per mosquito on day 13. Although virus titer in Cx. p. molestus was lower than in Cx. tritaeniorhynchus at the end of the experiment, this difference was not statistically significant. The median infective dose (ID50) for Cx. p. molestus was 2.83 log10PFU and for Cx. tritaeniorhynchus was 1.02 log10PFU per mosquito, and this difference also was not significant. There also was no significant difference between the median infective dose for transmission (TID50) per mosquito for Cx. p. molestus (5.34 log10PFU) and Cx. tritaeniorhynchus (4.59 log10PFU). We concluded that Cx. p. molestus is an effective laboratory vector of JE virus. PMID- 11004797 TI - Oral infection of Culicoides sonorensis (Diptera: Ceratopogonidae) by vesicular stomatitis virus. AB - Vesicular stomatitis virus serotype New Jersey (VSNJV) was mixed with bovine blood or fetal bovine serum (FBS) and fed across silicone membranes to laboratory populations of Culicoides sonorensis Wirth & Jones. In an initial study, virus was detected after 13 d in 21% of the midges that received an FBS/VSNJV mixture. In subsequent time-course experiments, engorged females were collected and maintained at 20.0 degrees C and assayed for VSNJV immediately after feeding and at 1, 3, 7, 10 and 13 d after feeding. Virus was detected after 13 d in 3% of the midges that received a bovine blood/VSNJV mixture and in 9% of the midges that received an FBS/VSNJV mixture. The results indicate that C. sonorensis should be considered as a potential biological vector of VSNJV. PMID- 11004798 TI - [Two-step tuberculin skin testing in our hospital employees]. AB - To evaluate the baseline values of tuberculin reaction, two-step tuberculin skin testing was carried out in 365 employees of our hospital. We defined strongly response group when the size of erythema showed more than 30 mm or who showed strongly positive reaction. Two-step tuberculin skin testing was carried out in 165 hospital employees excluding those who were defined as the strongly response group in the first testing. 80 hospital employees (48.5%) became strongly response group by the second tuberculin skin testing. Altogether, 76.7% of all employees were strongly response group either by the first time or the second tuberculin skin testing. The size of erythema and that of induration showed 13.2 +/- 12.6 mm (mean +/- SD), 6.9 +/- 9.2 mm increase, respectively, in the two-step tuberculin skin testing, so-called Booster phenomenon. We could not know the true tuberculin reaction status by the single tuberculin skin testing. Therefore, two step tuberculin skin testing is important as one of an infection prevention countermeasures in the hospital workers. PMID- 11004799 TI - [Transitional pattern of the clinical features of patients with pulmonary tuberculosis in a community hospital]. AB - To determine changes in the clinical features of recent patients with pulmonary tuberculosis in a community hospital without restricted tuberculosis wards, the clinical findings of 112 patients with pulmonary tuberculosis (containing miliary tuberculosis) during the past 15 years were compared by dividing the patients into three groups, each encompassing a five-year period. Recently, the number of patients with pulmonary tuberculosis was found to be increasing in a community hospital. In particular, the percentages of elderly patients and smear positive patients have increased. However, because of the improving awareness on tuberculosis, we have diagnosed TB cases correctly on admission and tended to perform the appropriate treatment. The comparative study between pulmonary tuberculosis patients diagnosed at the outpatients department and diagnosed after admission indicated that the patients diagnosed after admission showed pneumonia like infiltrative shadows without cavity formation and lower smear positivity for tubercle. Fortunately, resistance to antituberculous drugs of isolated tubercle bacilli in our community hospital has not yet increased and the prognosis of the cases proved to be good when the appropriate treatment was performed at an early stage. PMID- 11004800 TI - [Management of pulmonary tuberculosis patients complicated with diabetes mellitus before diagnosis as pulmonary tuberculosis and feasibility of chemoprophylaxis]. AB - We studied whether diabetics who are one of the high risk groups of developing pulmonary tuberculosis had undergone chest X-ray examination periodically. The feasibility of chemoprophylaxis in diabetics was also studied by investigating whether fibrotic lesions of tuberculosis can be found on previous chest X-ray films of these patients. Of the pulmonary tuberculosis patients admitted to our hospital for treatment, 78 patients complicated with diabetes mellitus were enrolled in this study. As to the mode of detection, the majority, 63 cases, are discovered, by undergoing medical examination because of respiratory symptoms, followed by 8 patients in whom pulmonary tuberculosis was found by health examination and only 1 patient was found by the periodic observation of diabetes mellitus. Of the 57 patients receiving the original treatment for pulmonary tuberculosis preceded by the discovery of diabetes mellitus, only 15 (26%) had undergone chest X-ray examination periodically. This fact shows that physicians treating diabetes mellitus have only a little concern on tuberculosis, thus the re-training of physicians dealing with diabetics on tuberculosis is considered to be necessary. According to chest X-ray films of 21 patients who had undergone chest X-ray examination and in whom the previous films were available, there were 6 patients without any lesion of pulmonary tuberculosis, 8 patients with fibrotic lesions and 7 patients with active lesions. The 8 patients showing fibrotic lesions have developed pulmonary tuberculosis on the average 15 years after they were diagnosed with diabetes mellitus, and the fact suggests that the prevention of the development of pulmonary tuberculosis among diabetics could be possible by chemoprophylaxis. PMID- 11004801 TI - [Study of the two-step tuberculin tests and gamma-interferon assays in elderly persons]. AB - Recently, an outbreak of lung tuberculosis among elderly residents happened in the home for aged, and it has become a social problem. To analyze the problem of tuberculosis in the elderly persons, we performed two-step tuberculin tests at two weeks interval and gamma-interferon assays for diagnosis of human tuberculosis to residents in the home for aged. 45 males (age; 77.3 +/- 8.9 SD) and 118 females (age; 81.9 +/- 8.6 SD) elderly persons (> or = 60) were examined. Standard dose of PPD was injected intracutaneously, and read at 48 hours. Erythema of 10 mm diameter or more was considered positive. The results were as follows. (1) The percentages of positive reactors in the 1st (T1) and the 2nd (T2) tuberculin tests were 50.3% and 68.1%, respectively. (2) The diameters of erythema and percentage of positive reactors (T1) did not very with age, but the size of erythema increased from 13.5 mm (T1) to 21.2 mm (T2) among those below 80 years, and from 13.0 mm (T1) to 16.4 mm (T2) among those 80 years or higher (p < 0.01). (3) The percentage of positive reactors (T1) varied by sex, namely males react stronger than females. (4) A dose of interferon-gamma released specifically in response to PPD stimulation of whole blood cultures showed no close relation with the diameters of erythema. In conclusion, tuberculin reactivity waned among elderly persons, and the fact suggests that they would have lower resistance against new infections, and as a result, they might have higher risk of developing tuberculosis. Elderly residents should be performed two-step tuberculin tests at the time of their entrance to the home for aged, and negative reactors should carefully be followed up. PMID- 11004802 TI - [Study on recombinant BCG]. AB - The progress of study on recombinant BCG was stated briefly. And then our studies on recombinant BCG were mentioned. Recombinant BCG secreting alpha antigen-fused merozoite surface protein 1 (MSP 1) was prepared and tested for its ability to control infections of Plasmodium yoelii. Result turned out it controlled the infection better than recombinant MSP 1 mixed with Freund incomplete adjuvant did. Recombinant BCG secreting excess amounts of antigen 85 complex A controlled infection of Mycobacterium leprae. Addition of recombinant BCG secreting alpha antigen-fused IL-2 to peritoneal exudate cells induced IFN-gamma resulting in killing bladder cancer cells more efficiently than parental BCG did. PMID- 11004803 TI - [Pathophysiology of reflux esophagitis]. AB - Reflux esophagitis is more frequent in developed countries, and its prevalence increased in Japan in these 1/4 centuries. These increase might be based on the increase in the population of the elderly, advances in endoscopic technology, increase in the interest on esophagitis, and declined prevalence of Helicobacter pylori infection. Main pathogenesis on the reflux esophagitis are classified as 2 components. One is the motility factor of the stomach and the esophagus. Distention of the stomach causes transient LES relaxation(TLESR), leading acid reflux into the esophagus. Delayed gastric emptying enhances gastric distention, causing TLESR. Resting LES pressure might be lower than normal. Hiatal hernia is also an exaggerating factor, causing repeated reflux from the hernia sac. The other main cause is the intact acid secretion causing enough to injure the esophageal mucosa. Gastric atrophy due to Helicobacter pylori infection would be protective factor for reflux esophagitis. PMID- 11004804 TI - [Clinical update of GERD]. AB - In our country, it is reported that reflux esophagitis(GERD) has been increasing like in European countries. One of the reasons for this tendency may be the arrival of the aging society. Another explanation is the lowering of Helicobacter pylori infection rate in the younger generation. Though the Los Angeles classification is mainly used at present, the Savary-Miller classification is also clinically important. For the initial treatment of GERD, proton pump inhibitor(PPI) has been mainly used, while there is large problems in the ideal way of maintenance therapy. The guideline on the GERD treatment is also being considered, based on EBM. PMID- 11004805 TI - [Gastric acid secretion in gastroesophageal reflex disease]. AB - The gastric acid secretion was evaluated by serum pepsinogen I/II ratio and serum Helicobacter pylori IgG antibody titer in gastroesophageal reflux disease(GERD) patient. GERD patients was 81 patients. Los-Angeles classification (LA)O, so called endoscopy negative reflux disease, was 6 patients, and LA-O-B patients was 68 patients. Helicobacter pylori infection ratio in GERD patients was 50.6%(41/81 patients). Pepsinogen I/II ratio in Helicobacter pylori positive GERD patients was no significant different from Helicobacter pylori negative GERD patients. We concluded that the gastric acid secretion in GERD patients was normal secretion. PMID- 11004806 TI - [Helicobacter pylori infection and eradication]. AB - H. pylori infection is associated with various gastroduodenal diseases such as gastritis, peptic ulcer, gastric cancer, gastric MALT lymphoma. H. pylori infection is suggested that it plays a role as protective factor not promoting factor for reflux esophagitis and GERD. Epidemiological studies showed lower prevalence of H. pylori infection in reflux esophagitis and Barrett's esophagus comparing the control. Increased occurrence of reflux esophagitis after curing of H. pylori infection was reported. However, the relationship between H. pylori infection and reflux esophagitis has not been actually made clear. Also the mechanism of reflux esophagitis occurrence after H. pylori eradication is not obscure. PMID- 11004807 TI - [Endoscopic classification of reflux esophagitis]. AB - A new endoscopic classification of reflux esophagitis was proposed in Los Angeles in 1994(LA classification). A mucosal break was defined as 'an area of slough or an area of erythema with a discrete lined demarcation from the adjacent or normal looking mucosa'. Reflux esophagitis is divided into four grades according to the severity of its mucosal break. We can find a minimal change during endoscopy which is not a mucosal break but is histologically or chromographically reflux esophagitis. I think Los Angeles classification must include the minimal change and I propose a modification of LA classification which includes the minimal change as grade M and the esophagus without any minimal change and mucosal break as grade N. PMID- 11004808 TI - [Histopathological diagnosis in reflux esophagitis]. AB - We reviewed the histopathological features for the diagnosis of reflux esophagitis and gastroesophageal reflux disease(GERD) including carcinogenesis of the esophagus. Histologically, the presence of capillary dilatation, elongation of papillary, hyperplasia of basal layer, inflammatory cells-infiltration, balloon cells in the epithelium, and ulceration were evaluated in GERD cases. Although, histopathological changes were not clear in endoscopic-negative GERD cases, immunohistochemical examination with cell cycle protein(PCNA, p21, and p27) revealed the same abnormalities with GERD cases. In Japan, the majority cases of GERD are evaluated in grade according to Los Angeles system, therefore the prevalence of Barrett's esophagus and cancer is very low. We hypothesize that esophageal squamous cell carcinoma arising from GERD different from Barrett's cancer sequence, and clinicopathological long-term follow up will be required to assess the carcinogenesis including gene analysis. PMID- 11004809 TI - [Clinical significance of 24-hour intraesophageal pH monitoring in GERD patients]. AB - 24-hour intraesophageal pH monitoring(24-pH-M) is a method of choice to measure the esophageal acid exposure, and evaluate a patient's lifestyle who has reflux symptoms. The 24-pH-M is indicated to the cases who have at least one of the following signs/symptoms; endoscopy negative GERD(gastroesophageal reflux disease), atypical clinical course for reflux disease, atypical symptoms of GERD, and those who are subjected to have surgical treatment. In the 24-pH-M, not only the duration of gastroesophageal reflux(GER), but also reflux and symptom association should be evaluated. The 24-pH-M is useful to investigate the diagnosis and treatment of GERD including reflux esophagitis. PMID- 11004810 TI - [Clinical significance of ambulatory intraesophageal bilirubin monitoring in diagnosis of gastroesophageal reflux]. AB - To evaluate the usefulness of ambulatory intraesophageal bilirubin monitoring in the diagnosis of GER, 38 patients with reflux symptoms and 7 controls were studied, using Bilitec 2000. The % time of bilirubin and acid reflux was less than 5% in all of the controls. In 3 patients of NUD, % time of acid reflux was less than 5%, but that of bilirubin was more than 8% in 2 patients. Acid reflux was found only in 1 of 4 patients who had a history of subtotal gastrectomy, but abnormal bilirubin reflux was found in all of 4 patients. In the other 31 patients, a significant difference was found in the % time of bilirubin reflux by the severity of reflux esophagitis, whereas this difference was not found in that of acid reflux. Intraesophageal bilirubin monitoring is useful in the diagnosis of GER and is essential in patients with hypochlorhydria or minimal reflux. PMID- 11004811 TI - [Esophageal peristalsis, lower esophageal function, and the methods of their evaluation]. AB - Esophageal peristalsis and lower esophageal sphincter(LES) function have an influence on gastroesophageal reflux disease(GERD). Incomplete contraction during primary and secondary peristalsis leads to poor clearance of refluxed gastric acid. Failure of LES function can result in a low basal LES pressure, absent or incomplete LES relaxation after swallowing, or an inadequate increase of LES pressure accompanying gastric activity. In addition, transient LES relaxation(TLESR) has been suggested as an important factor in GERD. Recent studies have indicated that TLESR has a relationship to nitric oxide(NO) and cholecystokinin(CCK). PMID- 11004812 TI - [The clinical evaluation of structured questionnaire for reflux esophagitis]. AB - Reflux esophagitis is suspected on clinical grounds and confirmed by a variety of endoscopical, roentgenological and physiological investigations, which are obviously inapplicable to an epidemiological survey. Questionnaire for the diagnosis of reflux disease(QUEST) can reduce bias introduced by observer variability, is practical, inexpensive, and noninvasive. QUEST also has good sensitivity and specificity for reflux esophagitis, so it's one of the useful diagnostic tools for reflux esophagitis. It, however, needs to be modified for the diagnosis of gastroesophageal reflux disease (GERD). Questionnaires to measure Quality of Life(QOL) quantitatively can be applied to the assessment of the severity of diseases or drug efficacy, and they are useful especially for the evaluation of endoscopy-negative GERD. PMID- 11004813 TI - [Guideline for treatment of gastroesophageal reflux disease]. AB - The endpoints should be settled before the treatment of GERD. These are symptom relief, healing and maintenance of erosive or ulcerative lesions, and also probably prevention of several complications of GERD. In medical treatment for GERD, we can take some different strategies, that is, single-agent, step-up or step-down therapy. The authors reviewed the effects of many drugs, being used for GERD therapy, and decided the levels of therapeutic effects from the view point of EBM. Using these results, we recommended that the therapy of GERD should be started with PPI of a regular dose and stepped-down in usual cases, but it should be begun with PPI and stepped-up in refractory cases. PPI may be necessary also for maintenance treatment in most latter cases. The guideline of the treatment for GERD were discussed. PMID- 11004814 TI - [Comparison between histamine H2-blocker and proton pump inhibitor on the effects of initial treatment in reflux esophagitis]. AB - I have reviewed the comparison between histamine H2 blocker(H2-RA) and proton pump inhibitor(PPI) on the effects of initial treatment in patients with reflux esophagitis. Pathophysiology of reflux esophagitis has been thought to be gastroesophageal reflux of the gastric acid, due to motor dysfunctions of the esophagus and the stomach. Endoscopic healing rate of reflux esophagitis is significantly higher 84% on PPI treatment than 52% on H2-RA treatment at 8 weeks period. Because suppression of the gastric acid secretion is significantly stronger and longer on PPI than H2-RA, especially this suppression is markedly found in day time and also night time. On the initial treatment of reflux esophagitis, PPI is choice so-called "step down therapy" after healing, H2-RA and/or prokinetic drugs are treated as maintenance therapy. PMID- 11004815 TI - [Drug therapy for nocturnal gastric acid breakthrough]. AB - Nocturnal gastric acid breakthrough(NAB) is defined as nighttime periods of over 1 hour duration, with a gastric pH < 4.0, during administration of a proton pump inhibitor(PPI). Although the causative mechanism of NAB is unclear, this phenomenon is a serious problem for patients with high grade reflux esophagitis. NAB is more frequently observed in patients with reflux esophageals, since this phenomenon is mainly observed in cases without H. pylori infection. Bedtime administration of H2 receptor antagonist rather than additional administration of PPI on the standard dose of PPI is effective for the control of NAB. PMID- 11004816 TI - [Maintenance therapy of mild form of GERD by H2 receptor antagonists]. AB - Because of high relapse rate after the healing by proton pump inhibitor(PPI) or H2 receptor antagonist(H2RA), GERD usually needs long time maintenance therapy. PPI is superior to H2RA in the first line as well as maintenance therapy. PPI is necessary for severe cases of GERD. However, H2RA is sufficient for milder form of GERD patients. Among the H2RA using in Japan, nizatidine has known to stimulate gastric emptying and elevate LES pressure. Nizatidine may be superior to other H2RAs in the treatment of GERD. Recently, nocturnal acid breakthrough which night time acid is secreted even PPI is administered twice daily has been documented. H2RAs are stronger than PPI to inhibit nocturnal acid breakthrough and may be better than night time acid reflux. PMID- 11004818 TI - [Surgical treatment for reflux esophagitis]. AB - Recently, PPI and laparoscopic Nissen operation were introduced to clinical field for the treatment of reflux esophagitis. The indication of surgical treatment for reflux esophagitis is as follows: (1) patients resistant to medical treatments, (2) free reflux patient, (3) patients with short esophagus or stenosis, (4) patients with respiratory complications. As for surgical treatment, there are many operative methods were reported. Recently, laparoscopic Nissen operation is the most popular procedure, and Toupet method, Dor method, and Tokai University method are employed for elderly patients or patients with sever esophagitis. Collis or Deitel operation is indicated for short esophagus patients. We should select appropriate operative method for individual patients considering about their pathophysiological condition. PMID- 11004817 TI - [Long-term maintenance treatment of reflux esophagitis resistant to H2-RA with PPI (lansoprazole)]. AB - 97 patients in 49 hospitals with erosive esophagitis unhealed after at least 8 weeks treatment with H2-RA were given primary treatment with 30 mg lansoprazol once daily. After 8 weeks of treatment with lansoprazol, 75(77%) patients were endoscopically healed. Healed patients were then given maintenance treatment with either 30 mg lansoprazol once daily, 15 mg lansoprazol once daily or 20 mg famotidine twice daily for 24 weeks. 86% of patients randomized to 30 mg lansoprazol, 70% of patients randomized to 15 mg lansoprazol were maintained in endoscopic healing throughout 24 weeks as compared with only 12% of patients randomized to famotidine in endoscopic healing. After 24 weeks of lansoprazol treatment basal gastrin levels were moderately increased. However no significant histopathologic lesion was found in the oxyntic gland mucosa. PPI(lansoprazol) was far superior to H2-RA(famotidine) in preventing recurrence of healed erosive esophagitis. A goal achieved without adverse events and significant abnormalities in the oxyntic mucosal exocrine or endocrine cell but a moderate increase in basal gastrine levels. PMID- 11004819 TI - [Simple antireflux measures for management of gastroesophageal reflux disease]. AB - It is widely accepted that proton pump inhibitors are the most effective therapy for management of gastroesophageal reflux disease. H2-receptor antagonists, prokinetic agents, and antacids are used in treatment of gastroesophageal reflux disease. In addition to these medical treatments, beneficial effects of simple anti-reflux measures on management of gastroesophageal disease are evaluated. Simple anti-reflux measures include elevation of the head of the bed, weight loss, reduced food intake, and the avoidance of precipitating foods and drugs. In this chapter, we summarized the utility of these simple anti-reflux measures for management of gastroesophageal reflux disease. PMID- 11004820 TI - [Cost-effectiveness of the treatment of reflux esophagitis: proton pump inhibitor versus histamine-2-receptor antagonist]. AB - Gastroesophageal reflux disease(GERD) is a common condition and acid-suppressing agents are the mainstays of treatment. A clinical decision analysis comparing a proton pump inhibitor(PPI), lansoprazole and a histamine H2-receptor antagonist (H2RA), ranitidine for the treatment of reflux esophagitis in Japan was performed using a Markov chain approach. The PPI was consistently superior to the H2RA with regard to both clinical effectiveness and cost-effectiveness. Prescription of PPIs for a one-month period would further enhance the cost-effectiveness of PPI treatment. The PPI first strategy is the preferred therapeutic approach for medical treatment of reflux esophagitis. We also recommend that prescription of PPIs for a one-month period be approved by the Japanese health insurance scheme. PMID- 11004821 TI - [Epidemiology of gastro-esophageal reflux disease]. AB - Gastroesophageal reflux disease(GERD) is believed to be common and chronic gastrointestinal disorder. Quantitative estimates of the actual prevalence of GERD are difficult to obtain, because those who seek health care probably represent only a tip of the iceberg of people with GERD. The prevalence of heartburn and/or regurgitation, considered to be reasonably specific symptoms for the diagnosis of GERD, ranged from 10% to 48% for heartburn, from 9% to 45% for regurgitation and 21% to 59% for both/either in systematic review of GERD and reflux esophagitis(RE). It is widely accepted that the prevalence is lower in Asian countries, including Japan, compared with that in Western countries. However, the prevalence of RE in Japan may be increasing. In the case of heartburn and regurgitation in GERD and endoscopic findings in RE, different understandings of these terms by different investigators and subjects may have contributed to the range of results. Symptom definitions and what is understood by them should be among the most important considerations when undertaking population prevalence studies on reflux like symptoms, to allow comparisons to be made between studies. PMID- 11004822 TI - [Aging-related physical and life style changes in the patients with reflux esophagitis]. AB - Incidence of reflux esophagitis(RE) has increased over the past 10 years in Japan, where aging of the population is rapidly progressing. The majority(73.6%) of the patients with RE consisted of non-elderly males having life style problems and of elderly females suffering from posture alterations. This suggested the importance of the disease onset among elderly female population in addition to that of the non-elderly male population. The risk factors specific to the elderly patients include not only persistent acid secretion and presbyesophagus, but also complication with orthopedic regression diseases with posture changes and osteoporosis. The association with the latter orthopedic regression diseases has been significantly increasing and is suggested to promote development of RE. Further increase in the prevalence of RE is foreseen in this aging-accelerating country. PMID- 11004823 TI - [Reflux esophagitis with hiatal sliding hernia]. AB - Hiatal sliding hernia is often found in aged patient and some of them contribute to the development of reflux esophagitis. We described the etiology of gastroesophageal reflux disease(GERD) briefly and mentioned the relationship between hiatal hernia and reflux esophagitis. The old patients suffers from GERD are accompanying hiatal hernia at the high rate. Our successful results of laparoscopic Nissen's operation were reported shortly. Furthermore, we reviewed the reflux esophagitis after distal gastrectomy as most important complication after surgery. We emphasize that we should not hesitate to perform the surgical treatment to the patients suffering from GERD with hiatal hernia or after gastrectomy because it is safe and bring them the better Q.O.L. after treatment. PMID- 11004824 TI - [Barrett's esophagus]. AB - The Barrett's esophagus showing columnar metaplasia upward to the esophagus from the esophago-gastric junction is one of the final appearance of reflux esophagitis and important as a precancerous state of esophageal adenocarcinoma. Especially the Barrett's mucosa with intestinal metaplasia has high potential risk for adenocarcinoma. Although the clinical definition of the esophago-gastric junction is not easy, the criteria of the esophago-gastric junction and the Barrett's mucosa proposed by the Japanese Society for Esophageal Diseases is useful. The gastro-esophageal reflux is important in the development of Barrett's mucosa not only in the classical Barrett's esophagus but also in the short segment Barrett's. PMID- 11004825 TI - [Collagen disease and reflux esophagitis]. AB - The incidence of reflux esophagitis in cases of collagen disease is about 10%. However, reflux esophagitis is seen in about 50% of scleroderma cases. Moreover, in scleroderma the severity is correlated with the time since onset of the disease. Endoscopy is performed more often in the severe cases, and little therapeutic effect can be expected without use of a proton pump inhibitor. PMID- 11004826 TI - [Gastroesophageal reflux and respiratory diseases]. AB - There is a relationship between gastroesophageal reflux and respiratory diseases. Reflux may trigger and aggravate respiratory diseases especially bronchial asthma. Both clinical and experimental observations suggest that the pathogenic mechanisms may be a vagal reflex and/or microaspiration of gastric acid into the trachea. General measures to avoid reflux and proton pomp inhibitor together with inhaled corticosteroid are the choice for treatment of asthma associated with gastroesophageal reflux. Xanthine derivatives and beta 2 adrenergic agonist might be withheld in these patients. A laparoscopic surgical treatment may be an option for patients with persistent symptoms. PMID- 11004827 TI - [Neurology]. PMID- 11004828 TI - [Eradication model employing Mongolian gerbils treated with chemical carcinogen and infected with Helicobacter pylori--implications for human cancer prevention]. AB - To explore the 'causal link' between Helicobacter pylori(Hp) infection and stomach carcinogenesis, we have established experimental models of stomach carcinogenesis in Mongolian Gerbils(MGs) using the chemical carcinogen, N-methyl N'-nitro-N-nitrosoguanidine(MNNG) and N-methyl-N-nitrosourea(MNU). The incidence of adenocarcinoma was significantly higher in animals treated with Hp than in the controls. Eradication of Hp was effective in reducing the incidence of adenocarcinoma. No gastric carcinoma was found without any chemical carcinogen. Hp infection enhances glandular stomach carcinogenesis not only of well differentiated type but also poorly differentiated type and signet ring cell type. Eradication of Hp, apparently a gastric promoter rather than gastric carcinogen, may be useful as a measure for prevention of stomach cancer. PMID- 11004830 TI - [Videofluorographic study of swallowing in patients with severe motor and intellectual disabilities--I. Recurrent pulmonary infections]. AB - We studied swallowing movements of 8 patients with severe motor and intellectual disabilities and recurrent pulmonary infectious diseases. All the patients were orally fed with no problems, but had frequent episodes of pyrexia, mostly due to pulmonary infections. Videofluorographic studies revealed severe impairments in the transitional and pharyngeal phases. The patients aspirated liquids, most frequently during swallowing. Prior to swallowing pooling of liquid barium was frequently observed in the hypopharynx, and that of purees in the pharynx after swallowing. The delay of cricopharyngeal relaxation was also frequently observed. Based on the videofluorographic findings, we made some efforts to provide patients with evident or latent aspiration with better management: changes in food textures and posture during drinking and eating. These efforts significantly reduced the frequency of pyrexic episodes. Since latent aspiration is not uncommon among patients with severe motor and intellectual disabilities, swallowing disturbance should be evaluated in detail by videofluorographic examination to improve QOL. PMID- 11004829 TI - [The protocol of clinical trial and basic experiments for esophageal cancer using gene transduction]. AB - We examined whether antitumor effect could be produced by retrovirally expressed interleukin-2(IL-2) gene, glanulocyte macrophage-colony stimulating factor(GM CSF) gene, herpes simplex virus-thymidine kinase(HSV-tk) gene and p53 gene in human esophageal cancer cells using nude mice. Loss of tumorigenicity of IL-2 or GM-CSF producing cancer cells were observed. The antitumor effect was also evidenced by the injection of these cells into established tumors of wild-type cells. In suicide gene therapy on esophageal cancer, the growth suppression of esophageal cancer cells transducing HSV-tk gene tumors in nude mice induced by ganciclovir treatment and all the tumors disappeared. The wild-type p53 transduced tumor cells became markedly susceptible to irradiation and anticancer agents. Administration of cisplatine noticeably suppressed the growth of p53 transduced tumors inoculated in nude mice. We established the clinical protocol of gene therapy for esophageal cancer using wild-type p53 gene with adenovirus vector. In this autumn we are going to start this clinical trial. PMID- 11004831 TI - [Videofluorographic study of swallowing in patients with severe motor and intellectual disabilities--II. Impairment of oral phase]. AB - We studied swallowing movements in 14 patients with severe motor and intellectual disabilities and development of the impaired oral phase. The oral phase in the patients did not develop beyond the late weaning phase. They had impairment of the oral phase: poor or absent lip seal, infantile swallowing, slowed or stalled bolus transit, and oral residue after swallowing. Videofluorographic studies revealed severe impairment in the transitional and pharyngeal phases: pooling of liquid barium in the hypopharynx prior to swallowing, the delay of cricopharyngeal relaxation, aspirated liquids (silent aspiration 77.8%), aspirated purees, pooling of purees in the pharynx after swallowing. After videofluorographic studies we made some efforts to provide patients with better management; changes in food textures and posture during drinking and eating. These efforts eased their feeding difficulties. Videofluorographic studies could be helpful in evaluating swallowing disturbance in the oral phase and thereby preventing chronic aspiration, malnutrition and feeding difficulties in patients with severe motor and intellectual disabilities. PMID- 11004832 TI - [Functional hemispherectomy for children aged 2 years or less for the treatment of intractable epilepsy caused by cortical dysgenesis]. AB - Thirteen children aged 2 years or less who had intractable epilepsy caused by cortical dysgenesis underwent functional hemispherectomy. The cerebral malformations were hemimegalencephaly in 8 cases and focal cortical dysplasia in 5 cases. Among 11 children who were followed for at least 6 months after the operation (6 to 54 months with a median of 26 months), 5 were seizure-free, 4 achieved > 90% seizure reduction, and 2 achieved 50-90% reduction. Ventriculo peritoneal shunt was placed in 3 children with hemimegalencephaly. After hemispherectomy, all the children showed improvement in psychomotor development. Development was accelerated in 3 seizure-free children. In children with cortical dysgenesis, functional hemispherectomy may result in remarkable seizure reduction and steady developmental progress. PMID- 11004833 TI - [A case of subacute sclerosing panencephalitis showing various cerebral perfusion on 99mTc-ECD SPECT]. AB - We reported serial neuroradiological findings of a 20-year-old male with subacute sclerosing panencephalitis, who deteriorated from Jabbour stage 2 to stage 4 rapidly in spite of oral administration of inosine pranobex. At the stage 2, brain magnetic resonance imaging (MRI), computed tomography (CT) and [99mTc]-L, L ethyl cysteinate dimer (99mTc-ECD) single photon emission computed tomography(SPECT) findings were normal. As the disease progressed, brain MRI and CT revealed diffuse cerebral white matter lesions as well as cerebral atrophy. 99mTc-ECD SPECT showed serially various perfusion of cerebrum. These findings in 99mTc-ECD SPECT may reflect the acute inflammatory process and consequent destruction of brain tissue. PMID- 11004834 TI - [Longitudinal study of cognitive function in two patients with focal cortical dysplasia]. AB - To clarify the relationship between epileptic attacks and cognitive dysfunction, we examined the serial findings of 123I-IMP single photon emission computed tomography (SPECT) in relation to the intelligence quotient (IQ), assessed by Wechsler Intelligence Scale of Children-Revised, in two female patients with focal cortical dysplasia (FCD) over a 10-year period. The age of patient 1 at the initial assessment was 2 years, and the age of patient 2 was 9 months. They developed complex partial epilepsy in infancy, and were treated with antiepileptic drugs, which remained effective until 11 years of age, when their epileptic attacks recurred. Patient 1, a 14-year-old girl with FCD of the left parietal lobe suffered from dyscalculia, right-left disorientation, and finger agnosia even when she was free of epileptic attacks. Following the recurrence of seizures which occurred every night, she became unable to understand what was said to her. A hypoperfusion area detected by 123I-IMP SPECT was restricted to the left parietal lobe during the seizure-free period, but spread to the temporo parietal lobes following the recurrence. Her verbal IQ declined from 94 (at 9 years of age) to 63 (at 11 years and 8 months). After her seizures were controlled again (at 14 years and 4 months), the 123I-IMP SPECT findings improved. Patient 2, a 12-year-old girl with FCD of the left frontal lobe, showed cognitive dysfunction. Her verbal IQ declined from 91 (at 7 years and 5 months) to 76 (at 11 years and 8 months) following a recurrence of epileptic attacks. 123I-IMP SPECT showed hypoperfusion in the left frontal lobe, where the accumulation count ratio (left/right ratio) declined from 0.86 (at 3 years) to 0.64 (at 11 years). These findings suggest that epileptic attacks are related to cognitive dysfunction in FCD patients. This cognitive dysfunction appears to correlate with the appearance of hypoperfusion areas, as detected by 123I-IMP SPECT. PMID- 11004835 TI - [Serum anti-arrestin antibody and disease activity of multiple sclerosis--a case report of 4-year-old child]. AB - We report a 4-year-old girl with multiple sclerosis (MS) and increased serum anti arrestin antibody. She was admitted to our hospital because of sustained fever, headache and anorexia. Serial magnetic resonance imaging (MRI) revealed diffuse high intensity of cerebral white matter and basal ganglia on T2-weighted imaging. We treated her with high-dose intravenous methylprednisolone under a diagnosis of ADEM. She recovered without any neurological sequelae in three months and the MRI lesion almost disappeared. However, she developed optic neuritis and two episodes of neurologic involvement associated with multiple white matter lesions in the next two years. She was then diagnosed as having clinically definite MS. Anti arrestin antibody is reportedly present specifically in adult patients with MS and uveitis. We performed an immunoblot assay using bovine retina, and our case revealed the presence of anti-arrestin antibody. Serial examinations showed that the serum anti-arrestin antibody titer correlated well with the disease activity. These results indicate that anti-arrestin antibody may be a good parameter of the disease activity of MS even in childhood. PMID- 11004836 TI - [Two cases of auditory disturbance caused by carbamazepine]. AB - Two cases of auditory disturbance caused by carbamazepine are reported. Patient 1, a 9-year-old boy with benign Rolandic epilepsy, developed impairment of pitch perception one day after administration of carbamazepine. Patient 2, a 33-year old female with glossopharyngeal neuralgia, developed impairment of pitch perception several hours after administration of carbamazepine, complaining that the perceived sounds became lower than previously by a semitone. Her pitch perception recovered soon after the cessation of carbamazepine. The carbamazepine induced disturbance in pitch perception may be more frequent than we guess. PMID- 11004837 TI - [Shuddering attacks in four children]. AB - Shuddering attacks (SA), an uncommon benign disorder occurring during infancy or early childhood, should be distinguished from epileptic seizures. The attacks are shivering movements occurring daily for several seconds without impairment of consciousness. SA are regarded as an early premature manifestation of essential tremor (ET), and the provoking mechanism is the same between them. There are case reports of SA, but none from Japan. We studied the pathophysiology of SA in four children aged between 8 and 14 months using a video-EEG monitoring system. In one patient, the shuddering movements, as indicated by contaminating electromyogram during electroencephalography, was almost as frequent as that of ET. SA decreased in frequency or disappeared in all our patients. One of them showed immature brain development on MRI and had relatives with epilepsy. Another showed flattened sella turcica. Although previous reports suggest that SAs are benign and require no investigation, children with SA could have borderline problems related to the development of the nervous system. PMID- 11004838 TI - [Three infants who suddenly became unwilling to stand up presumably due to acute cerebellar ataxia]. AB - We reported here three infants aged 11 to 12 months who, according to their parents, suddenly became unwilling to stand up. All the patients had barely achieved the ability to stand or to walk at the time of the illness. The symptom developed relatively acutely in the winter months from 1997 to 1998 with a probable preceding infection. On examination, no paralysis of the lower extremities was observed, and muscle weakness was minimal. There were neither violent tremors, nystagmus, nor sensory disturbance. Orthopaedic disorders were considered unlikely. Biochemical examinations of the serum were not diagnostic, and imagings of the central nervous system were normal. Spinal tap revealed mild cerebrospinal fluid pleocytosis in two of the patients. All the children showed almost complete recovery without any medication during the following period of 2 days to 2 months. Although no definite signs of ataxia were recognized in the three patients, a diagnosis of acute cerebellar ataxia was considered to be appropriate. Mild, infantile acute cerebellar ataxia may be overlooked because of the obscure complaints of the parents and the difficulty in performing a thorough neurological examination of infants. PMID- 11004839 TI - [Supplementary motor area epilepsy associated with ADHD in an abused history]. AB - A 6-year-old girl with attention-deficit hyperactivity disorder (ADHD) who had been abused by her mother in infancy developed supplementary motor area (SMA) epilepsy. The seizure was characterized by bilateral tonic seizure of the upper and lower extremities, speech arrest, preserved consciousness and a lack of postictal confusion. The duration of the seizure was usually 10-60 seconds. The seizures sometimes clustered. She was diagnosed as having SMA epilepsy based on the characteristic clinical symptoms, interictal EEG, ictal video-EEG and ictal SPECT. Though her seizure was initially improved by anti-epileptic drugs, the symptoms appeared again after discharge. Since her clinical course indicated that her seizure was aggravated by her mental state, treatment included both medication with anti-epileptic drugs and the adjustment of her living environment in cooperation with a child guidance clinic. Thereafter both her epileptic seizure and ADHD symptoms improved. These changes may be related to each other, because both conditions are associated with frontal lobe dysfunction. It was interesting that the adjustment of the environment improved frontal lobe epilepsy, which in turn ameliorated ADHD symptoms. PMID- 11004840 TI - [Historical review of pediatric brain death in Japan]. AB - Although brain death of children has recently been hotly discussed in Japan, there still remain uncertainty and confusion. A pediatrician's diagnosis that a child is brain-dead entails delicate and emotional issues. Circumstances of the patients, families and institutions are so variable that many questions cannot be answered. It is most important to be aware of these problems and to seek consensus in the community. Pediatricians should provide their best care to both the patients and their families. PMID- 11004841 TI - [Brain death in pediatric patients--from the viewpoint of pediatric intensive care]. AB - In Japan, "brain death" has been discussed exclusively in connection with organ transplantation. However, the concept of brain death, which emerged with the progress in intensive care medicine, should be discussed in the context of palliative care in the ICU. It should be recognized that intensive care medicine includes not only life-saving high-tech therapeutic modalities, but also ethical and psycho-social aspects of modern medicine. In order to find out a decent solution to pediatric brain death issues, it is essential to develop pediatric intensive care in Japan. PMID- 11004842 TI - [Role of nurses for donor's families: current issue of organ transplant in the United States]. AB - To define the roles of donor coordinators (certified procurement transplant coordinators; CPTCs), we visited University of Florida Colleges of Nursing and Medicine and certified procurement transplant interviewed (CPTCs) and clinical specialists in pediatric critical care. Recipients were dealt with certified clinical transplant coordinators (CCTCs). These coordinators are certified by taking examinations to see if they are equipped with specialized knowledge and communication skills. The small number of organs available for transplant poses a serious problem. In many cases, medical personnel are not proficient enough to recommend organ donations to the family members of potential donors. Thus, the role of CPTC includes education of health professionals. In one study, 95% of the donor family members felt that hospital staff members explained about transplantation well, while 67% of the nondonor family members felt the same. The concept of "brain death" was not fully understood by some of the latter. The timing and place of discussion as well as the skill levels of medical staffs (preferably CPTCs) were important in receiving the consent. It is necessary for the donor family members to understand the concept of "death". Communications between the hospital staff and family members are critical. In Japan, there is no solid foundation for pediatric organ transplantation. Specialized nurses in pediatrics are limited. Further, there is meager certification programs for CPTCs or CCTCs. Under the present state, nurses in ICU and pediatric wards may act as potential coordinators. Education of these nurses will be critical to increase the number of pediatric organ transplantation. PMID- 11004843 TI - [New possibilities for treatment of allergic inflammation in asthma]. PMID- 11004844 TI - [Pseudomonas aerogunosa pneumonia in patients treated at the Hospital for Chest Diseases]. AB - Retrospective analysis of pneumonia caused by Pseudomonas aeruginosa was made in 66 patients, treated in hospital. Nosocomial pneumonia was diagnosed in 11 (17%) patients. In 51 patients coexisting lung diseases were present: mainly COPD and bronchiectasis. Strains of Pseudomonas aeruginosa were susceptible mostly to imipenem, meropenem, aztreonam, ticarcillin-clavulanic acid, ceftazidime, ciprofloxacin, amikacin, piperacillin-tazobactam, netilmicin. Duration of treatment in hospital was very long--59% were treated over 30 days. Combined antibacterial therapy was applied in 35 (53%) patients and monotherapy, often with different antibiotics--in 31 (47%) patients. Treatment was successful in 45 (68%) patients. In 9 patients the results of treatment was not successful: mainly because of empyema in 7 pts. Twelve (18%) patients (with coexisting COPD--6 and lung cancer--6) died. We can support current recommendations for treatment of Pseudomonas aeruginosa infection with combination of aminoglycosides or fluoroquinolones plus one of remaining antipseudomonal antibiotics. Treatment failures occurred mainly in patients with severe coexisting diseases and/or empyema. PMID- 11004845 TI - [Cytoimmunologic changes in material obtained from bronchoalveolar lavage (BAL) in asymptomatic individuals chronically exposed to silica dust]. AB - The aim of this study was to determine and to evaluate silica induced lung cell reactivity--if any--in bronchoalveolar space, before clinical changes develop. Bronchoalveolar lavage (BAL) was carried out in 15 nonsmoking individuals with chronic professional silica exposure, free of lung signs and symptoms. Controls were healthy nonsmokers. Routine BAL cytology (HE, MGG) was completed by mast cell staining (toluidine blue). BAL lymphocyte subsets were phenotyped by direct two- and three-color immunofluorescence (applied DAKO A/S monoclonal antibodies: anti-CD3, CD4, CD8, CD11b, CD14, CD15, CD16 + 56, CD19, CD25, CD45, HLA-DR). Parallel staining was performed in peripheral blood. In individuals with chronic silica exposure we found: significant increase in alveolar macrophage (362 +/- 45 vs 160 +/- 33 x 10(3) cells/ml, p < 0.05), lymphocyte (61 +/- 9 vs 24 +/- 5 x 10(3) cells/ml, p < 0.05) and BAL total cell (415 +/- 76 vs 187 +/- 34 x 10(3) cells/ml, p < 0.05) numbers; significant increase in mast cell (0.4 +/- 0.1 vs 0.2 +/- 0.1, p < 0.05), NK cell (7.0 +/- 1.8 vs 3.6 +/- 1.0, p < 0.05) and Th early activated lymphocyte percent (CD4 + CD25+ calculated as percentage of CD4+ cells: 15.1 +/- 1.5 vs 7.8 +/- 1.6, p < 0.01). All results were presented as median +/- SEM. Bronchoalveolar space of people with chronic silica exposure usually shows pathological reaction (especially macrophagic alveolitis), although they are free of manifested pulmonary disease. Th early activated lymphocytes, NK cells and mast cells seem to play important role in the early interstitial lung tissue reaction to silica. PMID- 11004846 TI - [Proangiogenic cytokines (bFGF and VEGF) in BALF from two different lung segments examined by high resolution computed tomography (HRCT) in patients with sarcoidosis]. AB - The aim of the study was to evaluate the concentrations of bFGF and VEGF in double BAL (2 x 120 ml) from two different lung segments: (s.A) from upper lobe with the most and (s.B) from lower lobe with the least extensive involvement estimated by high resolution computed tomography (HRCT). Examined group consisted of 28 sarcoid patients with homogeneous, regular distribution of nodular opacities in conventional chest X-ray (14 F, 14 M aged 19-54). Eleven healthy volunteers served as controls. In patients with sarcoidosis we observed the significantly higher levels (p < 0.01) of bFGF (1.79 pg/ml, 1.48 pg/ml) and VEGF (107.5 pg/ml, 109.7 pg/ml) in BAL from s.A and s.B respectively in comparison with BAL from lung segments Abis and Bbis in control group (bFGF: 0.75 pg/ml, 0.47 pg/ml and VEGF: 33.7 pg/ml, 43.9 pg/ml respectively). bFGF in BAL from s.A in active sarcoidosis was higher than in s.A and s.B in non-active sarcoidosis. Concentrations of bFGF in BAL from both s.A and s.B correlated positively with CD4/CD8 ratio and absolute number of lymphocytes, CD4 cells and lymphocytes HLA DR estimated in BAL from these lung segments. We conclude that bFGF and VEGF may be involved in sarcoidosis pathogenesis and bFGF may be useful in estimation of sarcoidosis activity. PMID- 11004847 TI - [Detection of tubercle bacilli in clinical samples using genetic methods(PCR)compared with Lowenstein-Jensen culture methods]. AB - The aim of this work was to compare data obtained using PCR assay for detection of Mycobacterium tuberculosis with culture. We report a test for detection of tubercle bacilli by PCR and identification at species level by capture plate hybridisation and enzyme-linked immunoassay. 222 clinical samples obtained from patients with confirmed and suspected tuberculosis were analysed. These specimens were tested parallelly by conventional culture on Lowenstein-Jensen slants and PCR test. For 205 samples a complete agreement between these methods was observed. PMID- 11004848 TI - [Risk factors for airway obstruction in school children with respiratory symptoms assessed in a five year prospective study]. AB - The aim of this prospective study was to assess age-related changes of the respiratory status and to evaluate risk factors for persistent airflow obstruction in symptomatic school children. One hundred fifty children aged 7-12 years (mean age 9.5 yr.) were followed-up. In 1994 parents--administered questionnaire, skin prick tests and three dynamic spirometries in 4-6 week intervals were performed. Airways flow limitation was defined as FEV1%, MEF50 or MEF25 below minimal value. In 1999 questionnaires and spirometry were repeated by the same methods. The response rate was 74.7%. Children's respiratory status generally improved. Frequency of respiratory symptoms and obstructive disturbances (37.5% in 1994 versus 22.3% in 1999) decreased. Significant risk factors for airways flow limitation in 1999 were nocturnal dry cough, doctor diagnosed asthma, female gender and parental smoking. Advanced or persisting airway obstruction during several month in 1994 was valuable to predict lung function impairment in 1999. No relation between skin tests results and persistent airway obstruction was observed. PMID- 11004849 TI - [Sarcoidosis with central nervous system involvement]. AB - A case of pulmonary sarcoidosis with central nervous system involvement was described. Diagnosis was based on hilar lymph nodes biopsy and OUN MRI scan. Patient was treated with glicocorticosteroids with improvement. PMID- 11004850 TI - [Respiratory failure during the course of congenital myopathy effectively treated with nocturnal noninvasive nasal positive pressure ventilation]. AB - Respiratory muscle weakness in myopathy may result in respiratory failure. 41 year old male with congenital myopathy was successfully treated with nocturnal noninvasive nasal positive pressure ventilation. NPPV resulted in improvement of arterial blood gases and allowed to close tracheostomy. Patient was able to return to full time employment. PMID- 11004851 TI - [Bone tuberculosis in a patient with pulmonary tuberculosis]. AB - A case of bone tuberculosis in 83 years old farmer was described. He was hospitalized because of pain, oedema and purulent fistula in carpal regio. He hadn't neither systemic symptoms neither from respiratory tract. Chest X-ray and X-ray of carpal bones revealed tuberculous lesions. In the pus from cutaneous fistula AFB were found in culture. Antituberculous treatment was successful. PMID- 11004852 TI - [Genetic susceptibility to lung cancer]. PMID- 11004853 TI - [Lymphangioleiomyomatosis]. PMID- 11004854 TI - [Abnormal regulation of coagulation and fibrinolysis in lung cancer]. PMID- 11004855 TI - [Management of coagulation and fibrinolysis disorders in patients with cancer]. PMID- 11004856 TI - [Know the age of your lungs]. PMID- 11004857 TI - [A test for early detection of COPD]. AB - Free spirometry was offered to inhibitants of Bydgoszcz who presented with chronic cough. 1072 subjects were investigated, 638 males and 433 females, mean age 54 years. Eighty percent were smokers or exsmokers. In 78.6% subjects spirometry was normal. 200 persons (18.7%) had spirometric signs of airway obstruction, 19.5% mild, 51.5% moderate and 29% severe. Mass spirometry of smokers with chronic cough is a good screenings method for detection of COPD. PMID- 11004858 TI - [A test for initial diagnosis of COPD in patients with chronic cough and exercise dyspnea (population study)]. AB - The purpose of the present investigation was to evaluate the usefulness of spirometry as a screening test in the initial diagnosis of COPD in patients with chronic cough and exercise dyspnea. We encouraged people living in Lublin to the participation in this trial using local media. We examined 647 subjects, 329 (51%) females and 318 (49%) males between the ages of 14 and 89 years (average 55 y.). Among females there were: 73 smokers (22%), 51 former smokers (15%) and 205 non smokers (63%), among males respective numbers were: 112 (35.2%), 119 (37.4% and 87 (27.4%). 230 women (70%) and 230 men (72%) had clinical symptoms of COPD. The severity of bronchial obstruction was classified according to ERS recommendations. In females mild obstruction was found in 24 (7.3%), moderate in 80 (24.3%) and severe in 20(6.1%). In men mild obstruction was found in 15 subjects (4.7%), moderate in 80 (24.3%) and severe in 38 (11.9%). CONCLUSIONS: 1. Spirometry revealed that 35% of all examined subjects had an airway obstruction in majority of moderate and severe severity. 2. Spirometry is a very good and sensitive screening test in the diagnosis of COPD. PMID- 11004859 TI - [Early detection of COPD by high risk population spirometric screening]. AB - COPD is the most frequent chronic lung disease in Poland. The disease is however under-diagnosed, especially at the early stages. The aim of the study was to assess the efficacy of spirometric screening for COPD in middle aged smokers. Informations on causes and symptoms of COPD were disseminated in mass media in 14 large cities. Subject aged over 39 and with smoking history of > 10 packyears were invited for a free spirometry in local chest clinic. However, everyone attending had the spirometry performed. Spirometry was performed according to ATS recommendations. Airway obstruction (AO) was diagnosed when FEV1/FVC < 85% of N and categorised as mild (FEV1 > 70% of N), moderate (FEV1 50-69% of N) or severe (FEV1 < 50% of N). Spirometry was accompanied by an antismoking advice. RESULTS: 12.781 subjects were screened (mean age 52 +/- 12 years, 57% males). In 8.269 subjects who complied with inclusion criteria AO was diagnosed in 29.8% (mild in 10.9%, moderate in 12% and severe in 6.9%). In smokers < 40 years of age and a history of < 10 packyears AO was found in 8.8% (mild in 6.0%, moderate in 1.8% and severe in 1.0%). CONCLUSION: Mass spirometry is an effective and easy method for early detection of COPD. PMID- 11004860 TI - [Evaluation of the influence of tobacco smoking on pulmonary function in youg men]. AB - According to WHO estimates in the early 1990s there were around 1.100 million smokers in the world, about one third of the global population over 15 years of age. The aim of the study was to evaluate tobacco influence on pulmonary function tests in young healthy men. The study was performed on 3004 subjects 18-23 years old. The subjects were divided into two groups, smokers (S n = 1726) and non smokers (NS n = 1278). Each subject had spirometry and body pletysmography done. The analysis of influence of smoking on pulmonary function tests showed statistically significant decrease of TLC, FEV1, FEV1%/VC, PEF and FEF50 in the smokers' group. Percentage of people with airflow limitation (FEV1%VC < 85% N, FEV1 < 80% N, FEF50 < 70% N) was two times higher than in the non-smokers' group. IN CONCLUSION: smoking significantly increases the number of subjects with airflow limitation among healthy young males in Poland. PMID- 11004861 TI - [Comparison of automatic (AUTO-CPAP)and "manual" CPAP pressure titration in patients with obstructive sleep apnea]. AB - Auto-CPAP gives an opportunity to decrease costs of evaluating patient with OSA, replacing manual titration of pressure during PSG. The aim of this study was to compare automatic (auto-CPAP) and manual CPAP pressure titration in patients with OSA. We studied 50 obese patients (BMI--35 +/- 6 kg/m2), mean age 52.4 +/- 9.4 years with severe OSA, mean: AHI--62.9 +/- 22.1, mean overnight SaO2--89.1 +/- 3.7%, T90--54.4 +/- 29.6%. Two polysomnographies were performed: first when patient slept with CPAP and pressure was titrated manually by a technician and second on auto-CPAP device. Both methods had similar efficacy in reduction of AHI (< 10/h) and hypoxaemia, despite lower pressure established during auto-CPAP mode preventing apnoeas and hypopnoes during 90% of sleep time (8.2 +/- 1.7 cm H2O) compared to manual CPAP titration (9.2 +/- 1.7 cm H2O) (p < 0.05). CONCLUSION: Auto-CPAP seems to be a reliable alternative to manual titration of the therapeutic pressure in patients with OSA. This may help to cut a waiting list for PSG of patients suspected of OSA. PMID- 11004862 TI - [Obstructive sleep apnea and risk factors for coronary artery disease]. AB - STUDY AIM: Obstructive sleep apnoea (OSA) is strongly associated with obesity, especially abdominal obesity. Obesity in turn is a well-known risk factor for coronary artery disease (CAD). The aim of our study was to evaluate the relationship between OSA severity and CAD risk factors. MATERIAL AND METHODS: The sample consisted of 73 subjects (mean age +/- SE, 46.7 +/- 1 years) referred to a sleep laboratory. Subjects were either: 1. obese with OSA (O-OSA group n = 35; body mass index, BMI l 30 kg/m2; apnoea/hypopnoea index, AHI > 35), 2. non-obese with OSA (BO-OSA group n = 14; BMI < 27 kg/m2; AHI > 35), or 3. obese without OSA (O-Z group n = 24; BMI l 30 kg/m2; AHI < 5). All subjects underwent full overnight polysomnography. Blood samples were taken from all subject, for fasting levels of insulin (INS), glucose (GLU), total, HDL and LDL cholesterol, triglyceride (TG) and uric acid (UA). RESULTS: O-OSA had significantly higher INS and UA levels (p < 0.05) compared to BO-OSA and O-Z. GLU and lipid levels were comparable in the studied groups. GLU level correlated (p < 0.05) negatively to minimum oxyhemoglobin saturation (SAT-MIN) and positively to neck circumference. TG and UA levels were correlated (p < 0.05) positively to AHI and negatively to SAT-MIN. UA level was also positively correlated (p < 0.05) to BMI, waist/hip circumference ratio (WHR), and INS level. INS level correlated (p < 0.05) positively to AHI, T90, WHR and UA, and negatively to SAT-MIN and mean oxyhemoglobin saturation. After adjusting for the influence of OSA and obesity (multiple regression analysis), we found independent negative correlations (p < 0.05) between: GLU level and SAT-MIN, UA level and SAT-MIN, and INS level and SAT MIN. An independent, positive correlation (p < 0.05) was found between TG level and AHI. CONCLUSIONS: Results of our study suggest that OSA increases the risk of coronary artery disease by increasing plasma levels of glucose, triglyceride and insulin, independent of obesity. PMID- 11004863 TI - [Malfunction of some immunologic functions in patients with obstructive sleep apnea]. AB - Hypoxia and sleep disorders may result in an impairment of immune system in patients with obstructive sleep apnea (OSA). Therefore we studied mitogenic stimulation of peripheral blood lymphocytes by PHA (1 microgram/ml) by measurement of 3H-thymidine incorporation, metabolic activity of neutrophils measured by chemiluminescence (CL), immunoglobulins (IgG, IgA and IgM) concentration in serum (radial immunodiffusion) and TNF-alpha concentration in serum (ELISA), and E-rosette tests (with theophylline) in 57 obese patients with severe OSA. In OSA patients severe impairment of f-MLP-induced granulocyte chemiluminescent activity was observed. PMID- 11004864 TI - [Chronic Chlamydia pneumoniae infection in patients with asthma]. AB - The aim of our study was to evaluate the frequency of Chlamydia pneumoniae infection (especially chronic infection) in 41 asthma patients. They have been assigned to 3 groups, according to disease severity. Control group consisted of 35 age matched healthy volunteers (without cardiac and pulmonary diseases). The levels of specific IgG, IgA and IgM in patients' serum have been estimated using indirect microimmunofluorescence. According to serologic criteria, 23(56.1%) asthma patients and 4(11.4%) healthy controls have appeared to be chronically infected with Chlamydia pneumoniae (p < 0.001). Acute Chlamydia pneumoniae infection was present in 3(7.3%) asthma patients and in 2(5.7%) healthy controls. Taking in account asthma severity, persistent Chlamydia pneumoniae infection has occurred more frequently in patients with moderate and severe asthma than in ones with mild asthma. Acute Chlamydia pneumoniae infection was present in 9.1% and 12.5% of patients with mild and severe asthma respectively. PMID- 11004865 TI - [Chronic Chlamydia pneumonia infection in patients with chronic obstructive pulmonary disease]. AB - The aim of our study was to evaluate the frequency of Chlamydia pneumoniae infection (especially chronic infection) in COPD patients. Microimmunofluorescence method has been applied Chlamydia pneumoniae Micro-IF test (Labsystems) has been used. The levels of specific IgG, IgA and IgM have been estimated in patients' serum. According to serologic criteria, 64.1% of COPD patients and 20.5% of healthy controls appeared to be chronically infected with Chlamydia pneumoniae (p < 0.001). Taking in account COPD severity, persistent Chlamydia pneumoniae infection has been present in 68.2%, 57.1%, and 50% of patients with severe, moderate and mild COPD, respectively. Our study has revealed that chronic Chlamydia pneumoniae infection occurs more frequently in COPD patients than in healthy controls and in patients with severe COPD than in ones with mild and moderate disease. It is possible that persistent Chlamydia pneumoniae infection can initiate or amplify inflammatory reactions in the respiratory tract. The results suggest a need to diagnose chronic Chlamydia pneumoniae infection in COPD patients and, if confirmed, to take an attempt of antimicrobial therapy. PMID- 11004866 TI - [Lung changes during the course of non-Hodgkin's lymphoma in a patient with rhematoid arthritis with secondary Sjogren's syndrome]. AB - A case of 51 years old woman with 12-years history of rheumatoid arthritis and secondary Sjoegren's syndrome, who developed EBV-associated diffuse large B-cell lymphoma (Lymphomatoid granulomatosis type) involving lungs, lymph nodes and bone marrow was described. PMID- 11004867 TI - [Granulomatous lung lesions after occupational exposure to glass fibers]. AB - 39 years old man with granulomatous lesions in both lungs caused by occupational contact with glass fibers was described. He has been working as an bricklayer plasterer for 18 years and was in contact with lime, cement, plaster, asbestos, dust of coal and wood and with glass fibers. For the last two years before admission in 1993 he has had frequent bronchial infections. On admission he was in good general condition, his spirometric examination and blood gases were within normal limits. On chest x-ray disseminated lesions were found. Those lesions were of the round shapes on chest CT. Many sputum cultures for tubercle bacilli were negative. ANA and ANCA were not found in the serum. ACE was within normal limits. No precipitins to environmental antigens were found. Cancer metastases were suspected and lung biopsy during videothoracoscopy was done. Many foreign body type granulomas were found throughout the specimen. The character of the lesions was not typical for tuberculosis, sarcoidosis, extrinsic allergic alveolitis, silicosis or asbestosis. There are some reports concerning the possibility of development of such lesions after the exposition to glass fibers. We suspect that case is an example of such pathology. His occupational exposition was stopped in 1993 and he was observed without treatment. During the 5 years of observation (up till 1998) he was in good health with stable chest x-ray picture and results of respiratory system function. PMID- 11004868 TI - [The role of infection in chronic obstructive pulmonary disease]. PMID- 11004869 TI - [The role of anticholinergics in treatment of diffuse airway obstruction]. PMID- 11004870 TI - The proof of the pudding: an illustration of the relative strengths of null hypothesis, meta-analysis, and Bayesian analysis. AB - Some methodologists have recently suggested that scientific psychology's over reliance on null hypothesis significance testing (NHST) impedes the progress of the discipline. In response, a number of defenders have maintained that NHST continues to play a vital role in psychological research. Both sides of the argument to date have been presented abstractly. The authors take a different approach to this issue by illustrating the use of NHST along with 2 possible alternatives (meta-analysis as a primary data analysis strategy and Bayesian approaches) in a series of 3 studies. Comparing and contrasting the approaches on actual data brings out the strengths and weaknesses of each approach. The exercise demonstrates that the approaches are not mutually exclusive but instead can be used to complement one another. PMID- 11004872 TI - Coefficient alpha and related internal consistency reliability coefficients. AB - The author studied the conditions under which coefficient alpha and 10 related internal consistency reliability coefficients underestimate the reliability of a measure. Simulated data showed that alpha, though reasonably robust when computed on n components in moderately heterogeneous data, can under certain conditions seriously underestimate the reliability of a measure. Consequently, alpha, when used in corrections for attenuation, can result in nontrivial overestimation of the corrected correlation. Most of the coefficients studied, including lambda 2, did not improve the estimate to any great extent when the data were heterogeneous. The exceptions were stratified alpha and maximal reliability, which performed well when the components were grouped into two subsets, each measuring a different factor, and maximized lambda 4, which provided the most consistently accurate estimate of the reliability in all simulations studied. PMID- 11004871 TI - Effects of directionality of significance tests on the bias of accessible effect sizes. AB - The proportion of studies that use one-tailed statistical significance tests (pi) in a population of studies targeted by a meta-analysis can affect the bias of the sample effect sizes (sample ESs, or ds) that are accessible to the meta-analyst. H. C. Kraemer, C. Gardner, J. O. Brooks, and J. A. Yesavage (1998) found that, assuming pi = 1.0, for small studies (small Ns) the overestimation bias was large for small population ESs (delta < or = 0.2) and reached a maximum for the smallest population ES (viz., delta = 0). The present article shows (with a minor modification of H. C. Kraemer et al.'s model) that when pi = 0, the small-N bias of accessible sample ESs is relatively small for delta < or = 0.2, and a minimum (in fact, nonexistent) for delta = 0. Implications are discussed for interpretations of meta-analyses of (a) therapy efficacy and therapy effectiveness studies, (b) comparative outcome studies, and (c) studies targeting small but important population ESs. PMID- 11004873 TI - Large-sample confidence intervals for validity and reliability coefficients. AB - Large-sample confidence intervals (CI) for reliability, validity, and unattenuated validity are presented. The CI for unattenuated validity is based on the Bonferroni inequality, which relies on one CI for test-retest reliability and one for validity. Covered are four reliability-validity situations: (a) both estimates were from random samples; (b) reliability was from a random sample but validity was from a selected sample; (c) validity was from a random sample but reliability was from a selected sample; and (d) both estimates were from selected samples. All CIs were evaluated by using a simulation. CIs on reliability, validity, or unattenuated validity are accurate as long as selection ratio is at least 20% and selected sample size is 100 or larger. When selection ratio is less than 20%, estimators tend to underestimate their parameters. PMID- 11004874 TI - How important is transient error in estimating reliability? Going beyond simulation studies. AB - This article introduces a procedure for estimating reliability in which equivalent halves of a given test are systematically created and then administered a few days apart so that transient error can be included in the error calculus. The procedure not only estimates complete reliability (taking into account both specific-factor error and transient error) but also can estimate partial reliability (taking into account only specific-factor error). Scores from 6 different measuring instruments were analyzed with the procedure. The results indicate that the magnitude of transient error in real data can range from nonexistent to very large. It follows that traditional reliability estimates, using nonstaggered procedures, are inflated to the extent that transient error is present. PMID- 11004875 TI - Modeling stage-sequential change in ordered categorical responses. AB - Although few would dispute the usefulness of looking at behavioral change from a stage-sequential perspective, until recently the lack of appropriate modeling techniques has hampered rigorous empirical tests of stage theories. In particular, for behavioral measurements that are ordinal, there is a need for methods that represent the underlying change processes in the form of qualitative and discontinuous shifts. This article introduces a stage-sequential ordinal model by postulating that at any point in time there are a finite number of latent stages. Panel members may shift among these stages over time. The authors show that the stage-sequential model provides a general approach for both the analysis of ordinal time-dependent data and tests of various competing theories and hypotheses about psychological change processes. An analysis of a 5-year study concerning attitudes toward alcohol consumption by teenagers is presented to illustrate the modeling approach. PMID- 11004876 TI - Executive processing. PMID- 11004877 TI - Task switching and the measurement of "switch costs". AB - The measurement of "switch costs" is held to be of interest because, as is widely believed, they may reflect the control processes that are engaged when subjects switch between two (or more) competing tasks. [In task-switching experiments, the reaction time (RT) switch cost is typically measured as the difference in RT between switch and non-switch (repeat) trials.] In this report we focus on the RT switch costs that remain even after the subject has had some time to prepare for the shift of task, when the switch cost may be approximately asymptotic (so called residual switch costs). Three experiments are presented. All three experiments used Stroop colour/word, and neutral stimuli. Participants performed the two tasks of word-reading and colour-naming in a regular, double alternation, using the "alternating runs" paradigm (R. D. Rogers & S. Monsell, 1995). The experiments were designed to test the hypothesis that RT switch costs depend on a form of proactive interference (PI) arising from the performance of a prior, competing task. A. Allport, E. A. Styles and S. Hsieh (1994) suggested that these PI effects resulted from "task-set inertia", that is, the persisting activation suppression of competing task-sets, or competing task-processing pathways. The results confirmed the existence of long-lasting PI from the competing task as a major contributor to switch costs. Non-switch trials, used as the baseline in the measurement of switch costs, were also shown to be strongly affected by similar PI effects. However, task-set inertia was not sufficient to account for these results. The results appeared inconsistent also with all other previous models of task switching. A new hypothesis to explain these between-task interference effects was developed, based on the stimulus-triggered retrieval of competing stimulus-response (S-R) associations, acquired (or strengthened) in earlier trials. Consistent with this retrieval hypothesis, switch costs were shown to depend primarily on the S-R characteristics of the preceding task (the task that was switched from) rather than the upcoming task. Further, the effects of the other, competing task were found to persist over many successive switching trials, affecting switch costs long after the stimulus overlap (and hence the principal S-R competition) between the current tasks had been removed. Switch costs were also found to be affected by recent, item-specific experience with a given stimulus, in either the same or the competing task. Finally, the results showed that switch costs were massively affected by the ratio of the number of prior trials, in response to the same stimuli, that had implemented either the currently intended or the competing S-R mappings. None of these effects are predicted by current models of residual switch costs, which appeal to the differences in control processes assumed to be engaged in switch versus non switch trials. PMID- 11004878 TI - Modeling cognitive control in task-switching. AB - This article describes a quantitative model, which suggests what the underlying mechanisms of cognitive control in a particular task-switching paradigm are, with relevance to task-switching performance in general. It is suggested that participants dynamically control response accuracy by selective attention, in the particular paradigm being used, by controlling stimulus representation. They are less efficient in dynamically controlling response representation. The model fits reasonably well the pattern of reaction time results concerning task switching, congruency, cue-target interval and response-repetition in a mixed task condition, as well as the differences between mixed task and pure task conditions. PMID- 11004879 TI - Reconfiguration of task-set: is it easier to switch to the weaker task? AB - Switching between two tasks afforded by the same stimuli results in slower reactions and more errors on the first stimulus after the task changes. This "switch cost" is reduced, but not usually eliminated, by the opportunity to prepare for a task switch. While there is agreement that this preparation effect indexes a control process performed before the stimulus, the "residual" cost has been attributed to several sources: to a control process essential for task-set reconfiguration that can be carried out only after the stimulus onset, to probabilistic failure to engage in preparation prior to the stimulus, and to two kinds of priming from previous trials: positive priming of the now-irrelevant task set and inhibition of the now-relevant task-set. The main evidence for the carry-over of inhibition is the observation that it is easier to switch from the stronger to the weaker of a pair of tasks afforded by the stimulus than vice versa. We survey available data on interactions between task switching and three manipulations of relative task strength: pre-experimental experience, stimulus response compatibility, and intra-experimental practice. We conclude that it is far from universally true that it is easier to switch to the weaker task. Either inhibition of the stronger task-set is a strategy used only in the special case of extreme inequality in strength, or its consequences for later performance may be masked by slower post-stimulus control operations for more complex tasks. Inhibitory priming may also be stimulus specific. PMID- 11004880 TI - Rehabilitation of executive deficits in closed head injury and anterior communicating artery aneurysm patients. AB - This paper reports a study that was aimed to rehabilitate executive functions in closed head injury (CHI) and anterior communicating artery (ACoA) aneurysm patients. The groups tested comprised 10 CHI patients, 9 ACoA aneurysm patients and 19 controls. We employed a dual-task paradigm that is known to tap the ability to co-ordinate two actions. The treatment consisted of five experimental sessions, in which the dual-task paradigm was used. In the CHI study, the dual task cost was measured before the treatment (assessment), immediately after the treatment (retest), and 3 months after the treatment (follow-up). In the ACoA aneurysm study, the dual-task cost was also assessed 12 months after the treatment. A significant reduction of the dual-task cost from assessment to retest was found. This reduction remained stable in the follow-up sessions. The results are discussed with reference to the absence of spontaneous recovery of this specific executive function and to the possibility that the beneficial effect of the treatment generalises to other executive functions and/or daily living activities. PMID- 11004881 TI - Strategy application disorder: the role of the frontal lobes in human multitasking. AB - Strategy application disorder is a term used to describe a pattern of deficits, usually associated with frontal lobe dysfunction, where people show disorganisation, absentmindedness and problems with planning and decision making in everyday life despite normal performance on traditional neuropsychological tests. It is argued that the prototypical situation which presents problems for these cases are those which require multitasking, and although good cases are rare in the literature, those that do exist show a characteristic neuropsychological pattern. Moreover, this pattern is confirmed in recent group studies of multitasking and of the relationship between multitasking tests (such as the Six Element Test), failures in everyday life and other neuropsychological measures. At present the evidence suggests that the potential frontal brain regions most implicated in multitasking are the anterior cingulate; B.A. 10 and immediately adjacent areas; and the right dorsolateral prefrontal cortex, with each making a unique contribution to different aspects of performance. Furthermore, recent studies show striking dissociations between performances on multitasking tests and two of the most commonly administered measures of executive function: the verbal fluency test and the Wisconsin Card Sorting Test, which sets a minimum level for a fractionation of the executive syndrome in humans. PMID- 11004882 TI - Executive functions and the frontal lobes: a conceptual view. AB - Several problems in understanding executive functions and their relationships to the frontal lobes are discussed. Data are then presented from several of our studies to support the following statements: (1) the examination of patients with focal frontal lobe lesions is a necessary first step in defining the relation of executive functions to the frontal lobes; (2) there is no unitary executive function. Rather, distinct processes related to the frontal lobes can be differentiated which converge on a general concept of control functions; (3) a simple control-automatic distinction is inadequate to explain the complexity of control-automatic processes; (4) the distinction between complex and simple tasks cannot explain the differences in functions between the frontal lobes and other brain regions; and (5) the most important role of the frontal lobes may be for affective responsiveness, social and personality development, and self-awareness and unconsciousness. PMID- 11004883 TI - Neural control of behavior: countermanding eye movements. AB - Understanding the self-control of action entails knowledge about how actions are initiated, how planned actions are canceled and how the consequences of actions are registered. We have investigated neural correlates of these processes using the countermanding paradigm--a task that required subjects to occasionally cancel a planned speeded response, and an analysis that provides an estimate of the time needed to cancel a planned movement. By monitoring the activity of single neurons in the frontal cortex of macaque monkeys performing this task we have distinguished signals responding to the visual stimuli, other signals that control the production of movements, and still other signals that seem to monitor behavior. PMID- 11004884 TI - Patterns of cognitive ageing. AB - Neuroanatomical evidence suggests that normal ageing affects some brain areas, and the "local" functions they support, earlier and more severely than others. Changes appear to be especially marked in the hippocampus, temporal association and prefrontal cortex. Evidence from classical neuropsychological studies suggests that these brain areas are associated with memory and "executive" functions, respectively. We may, therefore, expect that tests purported to measure these functions may be disproportionately affected in old age and that there may be evidence for some separation of these functions even within neurologically normal populations. What we also know, however, is that measures reflecting general fluid ability also decline with increasing age, so any hypothesis relating to specific "local" deficits must acknowledge and account for any "globar" changes in performance. Volunteers (n = 162) aged between 60-80 years who had completed the Cattell and Cattell Culture Fair Intelligence Test (CCF) completed the Cambridge Automated Neuropsychological Test Battery (CANTAB). The CANTAB has been administered to several patient populations and tests from the battery have been shown to be sensitive to damage in both temporal and prefrontal areas (Owen et al., 1996). Results from the test battery showed that both the Paired Associate Learning and Spatial Recognition tests were the most sensitive to normal ageing even when CCF is accounted for. In contrast, this performance on the "executive" tests, shown to be sensitive to frontal lobe damage was not related to age, and CCF scores predicted performance on these tests. These results are discussed in relation to theories of cognitive ageing and patterns of change and in relation to several important methodological and theoretical considerations for the study of executive function. PMID- 11004885 TI - [The "international" of psychodynamic psychotherapy researchers]. PMID- 11004886 TI - [Illness-coping in a population-based representative sample. Situational, sociodemographic and social influences]. AB - In spite of the considerable volume of current coping research there is a noticeable lack of studies based on large representative samples. In this study a sample of 2179 subjects of 16 to 96 years of age that was representative of the entire population of Germany, was asked as to how they now cope, how they have coped in the past or how they would cope with: 1. an existing illness, 2. a past illness and 3. an imagined (non-existent) illness. The instrument used was the questionnaire version of the "Berner Bewaltigungsformen" (BEFO), a coping questionnaire with 28 systematically derived coping dimensions that are defined independently of the usual empirical-topographical dimensions (behavioural, cognitive and emotional) of the type of illness. The enquiry was concerned with the influence of varying situational (proximity of illness, subjective evaluation of personal health, psychological symptoms), socio-demographic (age, gender) and social (social support) variables on the choice of coping mechanisms. The results showed that each of these factors influences the repertoire of coping mechanisms to varying degrees. This systematic variability in a representative sample should be taken into account in any evaluation of coping with illness in clinical sub groups. PMID- 11004887 TI - [Results of an evaluation of psychotherapeutic care from the viewpoints of patients and therapists in Saxony-Anhalt]. AB - With the enforcement of the German law for psychotherapeutics at the beginning of 1999, the field of psychotherapy detached from a widespread "delegation procedure" through the possibility of first-access for patients to a psychotherapist. In the form of an experiment, Saxony-Anhalt practiced as the only land in the Feral Republic of Germany a modified form of delegation procedure in the legal health insurance since 1992. The most important modification consisted in the possibility of first-access for the patient to the psychotherapist. Using selected results of the investigation in the view of psychotherapy patients and from medical and psychological psychotherapists, the situation after 5 years since introduction of this model in the out-patient, psychotherapeutic care in Saxony-Anhalt will be shown, as well as experience gained from this model. PMID- 11004888 TI - [Self concept and object relations of patients with affective disorders- individual centered diagnosis with the repertory-grid technique]. AB - Significant dimensions of the self concept and the object relations of 127 depressive patients and 34 orthopaedic patients were investigated with the repertory-grid-technique. Self concept and object relations were compared by means of nomothethically used idiographic results after recovery from manifest depression. The object relations of the depressive sample were characterised by the dimension "symbiotic near"; "ambivalent" and "indifferent" partnership relationships were found much more frequently in the controls. The idiographic results contribute to a differentiation of the spectrum of affective disorders. They underline the importance of the interpersonal dimension of depression and may be used as a basis of a therapeutic orientation. PMID- 11004889 TI - [Catamnestic study of ecological brief therapy]. AB - Ecological brief psychotherapy has been practised at the Psychiatric Outpatient Clinic at the University Hospital Zurich since 1990. The aim is to use the development-oriented challenges posed by important others in the therapeutic process. The effectiveness of this form of therapy is investigated by reviewing the outcome of 28 completed treatment cases. While results at completion of therapy come up to the expectations, the long-term results were surprisingly positive. 96.4% of those surveyed reported they felt considerably better at final assessment 1 to 6 years after the therapy than they did at the beginning. Positive changes continued after the completion of therapy. We presume that our coevolutive focus brought resolution of blocked developmental steps in relationships that further stimulated changes that continued after the conclusion of therapy, even without further therapeutic support. Furthermore, the treatment effects provided by ecological brief psychotherapy apply to personality disorders as well. PMID- 11004890 TI - [Rabies-free status of Switzerland following 30 years of rabies in foxes]. AB - The European fox rabies epizootic starting in 1939 at the eastern border of Poland reached Switzerland on March 3, 1967. Rabies spread over large parts of the country until 1977, the year it caused three human deaths. In 1978 the first field trial world-wide for the oral immunization of foxes against rabies was conducted in Switzerland. Initially, the expansion of the vaccination area led to a rapid reduction in rabies cases. However, the 1990s were characterized by a recrudescence of rabies in spite of regular oral immunization of foxes. The last endemic case of rabies was diagnosed in 1996 after an adaptation of the vaccination strategy. A total of 17,109 rabies cases, of which 73% in foxes and 14% in domestic animals were diagnosed, leading to an estimated number of some 25,000 postexposure treatments in humans. To eliminate rabies, a total of 2.8 million baits containing a modified live virus were distributed--mostly by hand- in the field. PMID- 11004891 TI - [The development of rabies in Switzerland--landscape determines the spread of a wildlife epidemic]. AB - The rabies epidemic that reached Switzerland in 1967 developed in response to landscape factors as long as no efficient control strategies were available. The landscape acted either as barrier to the spread of rabies, or it influenced the density of red foxes and thus the habitat of the epidemic. Following the first cases in the canton of Schaffhausen, the whole northwestern Switzerland was infected followed by the eastern Alps, large parts of the Plateau and the Jura mountains. In 1978, in the canton of Valais, the first campaigns of oral immunization of foxes against rabies started. The design of vaccination campaigns during the next two decades was always closely linked to landscape features. Thus, it was possible to free first the Alps and then the Plateau from rabies and finally, at the end of the 1990s, to eliminate it completely within the country. We describe the entire development of the epidemic within the period of 30 years from the first infection up to the last registered case and the final vaccination campaign. PMID- 11004892 TI - [The development of strategies for the field application of oral immunization of foxes against rabies]. AB - Preconditions for successful field application of the idea to orally immunize foxes against rabies were a) the development of an efficacious and safe vaccine and an attractive bait and b) a suitable strategy to reach a high proportion of the vector population with vaccine-ladden baits. A geographical and epidemiological concept on how to implement the vaccination strategy and how to achieve a sufficiently high level of surveillance were further cornerstones contributing to the success of the method. Finally, political decisions and a legal basis for the application of the method of oral immunization were required to eradicate fox rabies in Switzerland. PMID- 11004893 TI - [The final stage of rabies in Switzerland]. AB - Since summer 1989, rabies in Switzerland has been restricted to the Jura Mountains in the north-west of the country. Even there, the last endemic focus disappeared in 1990, but a re-infection in the same year caused a new flare-up of the epizootic. Until 1994, the number of rabies cases increased again to 225. Control measures were intensified with doubled vaccination campaigns, increased bait densities, and additional vaccination campaigns to immunize young foxes at the den. As a consequence, the number of cases dropped to 25 in 1995 and to 6 in 1996. On December 21, 1996, the last endemic case of rabies in Switzerland was registered. After two years of continuing vaccination campaigns and surveillance, Switzerland became officially rabies-free at the beginning of 1999. In the present paper, we analyse the final stage of the epizootic. The re-infection in 1990 was caused by infected foxes immigrating from France, but as the immunization of the fox population in Switzerland was insufficient, the disease became again endemic immediately. The lacking herd immunity was partly a consequence of problems related to the vaccination system and even more of the rapid increase of the fox population. PMID- 11004894 TI - Oral immunization against rabies: afterthoughts and foresight. AB - The article contains personal views on some issues that are frequently addressed in discussions about rabies control, and on some related topics that are often overlooked. The first field applications of oral wildlife rabies immunization in the Swiss Rhone Valley were preceded by many years of international cooperative studies on efficacy and safety. They were significant "faits accomplis" that facilitated similar endeavors in other countries. Some aspects of the residual pathogenicity of oral rabies vaccines are discussed. The field efficacy of oral wildlife immunization is the outcome of complex interactions between vaccine and bait attributes, bait distribution procedures, and habitat properties. Significant difficulties hinder the interpretation of field observations on efficacy. Though oral wildlife immunization is not an animal welfare act and not a conservationist achievement, it is an attempt at zoonosis control intended to protect human health and prevent economic losses. PMID- 11004895 TI - [The Caucasus: recurrence of rabies in Georgia]. PMID- 11004896 TI - [The last case of canine rabies in Switzerland]. PMID- 11004897 TI - MedBytes. PMID- 11004898 TI - Certifying death. PMID- 11004899 TI - Death by chocolate? PMID- 11004900 TI - Negotiating with insurers. PMID- 11004901 TI - The gene machine. PMID- 11004902 TI - A physician's guide to the human genome. PMID- 11004903 TI - Presidential health. PMID- 11004904 TI - Differential diagnosis and management of an infant presenting in shock with a history of sickle cell anemia and a recent fall. AB - The presentation of shock in an infant can be subtle, yet must be recognized and treated very quickly to prevent decompensation and cardiopulmonary arrest. Treatment must begin as soon as shock is noted and before or along with the evaluation to establish the etiology of the shock. This case report illustrates these principles by describing an infant with sickle cell anemia who presented to the emergency department in shock after sustaining a fall. PMID- 11004905 TI - [The immunological basis of the administration of DTP-polio vaccine]. AB - The main subject of this essay is the analysis of the immunizing properties and possible side effects of DTP (whole cell)-polio vaccine, which since 1964 is the backbone of the immunization programme in the Netherlands. The concept, that the basis of the immunity conferred by diphtheria and tetanus toxoid and inactivated poliovirus is humoral, is now generally accepted. Assessment of the potency of these three components is based on their binding properties to specific immunoglobulines in standard-antisera. An example is the testing of neutralising antibodies against the three types of poliovirus in sera obtained from different groups of infants injected with increasing doses of this vaccine. Side-effects after injecting diphtheria- and tetanus toxoids and three types inactivated poliovaccine are negligible. For the pertussis component of the vaccine, this is not the case. The standardization of the potency of cellular pertussis vaccine is still based on the analysis of the protective capacity of a number of plain vaccine lots in the intercerebral mouse-protection test relative to the protection they confer in children. An international standard vaccine is available for this purpose, and the potency of individual lots can be expressed in International Units. At present acellular pertussis vaccines are also available for immunization against this disease. The components of such vaccines are at least for a number of components, arbitrarily chosen. Recently in a trial in Sweden, a five-component pertussis vaccine has given a similar protection of infants as a cellular vaccine of British origin. In addition this acellular vaccine induced less febrile reactions. However, acellular vaccines do not protect mice against intracerebral infection. because producers have decided to omit the "Outer Membrane Complex (OMC) as a component of the vaccine. In addition it has recently been shown that the presence of the cellular pertussis component in DPT vaccine will almost completely suppress tetanus antitoxins in the IgE fraction of the serum. This observation could indicate that this suppression is the result of a cellular immunity reaction by components in the bacteria. PMID- 11004906 TI - [Vaccination policy in Flanders]. AB - In the recent past (1997) the Flemish authorities have turned the vaccination policy into one of the top priorities of their health policy. Following the WHO guidelines they have brought forward a health objective anticipating a significant increase of the vaccination rate for a number of infectious diseases by 2002. Recent research shows that attention should be paid especially to measles, mumps and rubella (83% vaccination rate at 18-24 months), hepatitis B (74-69%) and type b haemophilus influenzae (78%). The vaccination rate in Flanders for polio, diphtheria, tetanus and whooping cough is good or adequate at the age of 18-24 months. In 2000, the Flemish authorities reserve about BEF 80 million for the purchase of basic vaccines distributed for free through their health inspections, just like in the previous years. In order to realise this objective, all medical bodies carrying responsibility in this field are appealed to. Moreover, the local regional consultative bodies, financially supported by the Flemish Community, may be expected to play a stimulating role in, so far, four out of the five Flemish provinces. A strong management from a central Flemish vaccination umbrella organisation is considered to be indispensable in order to have the vaccinating bodies deliberate, have them build a consensus and motivate them. This should result in clear and generally accepted vaccination schedules and a well-informed and sensitised population. A project in view of a rapid and efficient registration of vaccination data in a central Flemish vaccination database, accessible for vaccinating doctors is now running with 'Child and Family' and can subsequently be extended to the entire target group. PMID- 11004907 TI - [Diabetes mellitus type 1 and associated organ-specific autoimmunity]. AB - Autoimmune type 1 diabetic patients show a high prevalence of thyroid peroxidase (TPO), parietal cell (PCA), anti-adrenal (AAA) and anti-endomysium antibodies (EmA-IgA), which may be accompanied with clinical disease. We studied the grade of associated organ-specific autoimmunity and the pattern of prevalence of TPO and PCA by age, gender, duration, age at onset of diabetes, and HLA DR haplotype in 783 type 1 diabetic patients, consisting of 286 children and 497 adults (M/F: 389/394), with a mean diabetes duration of 11.8 +/- 10.1 years. The relationship between islet cell (ICA), glutamic acid decarboxylase-65 (GADA) and thyro-gastric auto-antibodies was also investigated. TPO were present in 21.6%, PCA in 18.3%, AAA in 2.2% and EmA-IgA in 2.1% of the patients. The presence of TPO is determined by gender (p < 0.0001), age (P = 0.0008), and PCA status (p = 0.029). The presence of PCA is only influenced by age (p = 0.0027) and TPO status (p = 0.0155). Patients with ICA+ > or = 3 years had a higher prevalence of thyro gastric auto-antibodies (p = 0.045) than ICA- subjects. Also, PCA were more prevalent in GADA+ than GADA- patients (p = 0.005). We observed an association between HLA DR5 and PCA (p = 0.0012). Dysthyroidism was more prevalent in TPO+ than TPO- subjects (p < 0.0001). PCA+ subjects had a higher prevalence of iron deficiency anaemia (p = 0.0099) and pernicious anaemia (p < 0.0001) than PCA- patients. In conclusion, particularly type 1 diabetic patients with persisting ICA > or = 3 years or with GADA, show a high prevalence of thyro-gastric auto antibodies. Based on antibody-positivity we observed a high prevalence of thyroid disease, iron deficiency anaemia and pernicious anaemia, which can compromise the health of the diabetic patient. PMID- 11004909 TI - [DMW special topic issue: Nephrology]. PMID- 11004908 TI - [Targeting of chlorine E6 by EGF increasing its photodynamic activity in selective ways]. AB - Certain tumor cells, such as squamous carcinoma cells, express an increased number of epidermal growth factor (EGF) receptors. The goal of this study was the targeted delivery of the photocytotoxic compound Sn(IV)chlorine e6 monoethylenediamine++ (SnCe6(ED)) to tumors that overexpress the EGF receptor. Therefore EGF was conjugated to SnCe6(ED) through a carrier, such as dextran (Dex) and human serum albumin (HSA), followed by the evaluation of the photocytotoxicity on the EGF receptor overexpressing MDA-MB-468 cell line. The photobiologic activity of these conjugates was then compared to a conjugate of the photosensitizer to HSA or dextran, or to the photosensitizer alone. In contrast to EGF-HSA-SnCe6(ED), the affinity of EGF for its receptor was substantially impaired upon conjugation in EGF-Dex-SnCe6(ED). In correlation with these results, EGF-HSA-SnCe6(ED) displayed a high cytotoxicity (IC50, 63 nM) on MDA-MB-468 cells at a light dose of 27 kJ/m2, whereas EGF-Dex-SnCe6(ED) showed very limited photocytotoxicity. In the presence of a competing EGF concentration (10 microM), EGF-HSA-SnCe6(ED) was not cytotoxic anymore. The high photocytoxicity of EGF-HSA-SnCe6(ED) was shown to be a result of a high intracellular concentration in MDA-MB-468 cells, which could be lowered dramatically by incubating the conjugate with a competing EGF concentration. In contrast, EGF-Dex-SnCe6(ED) displayed very poor accumulation in MDA-MB-468 cells, in agreement with its low EGF receptor affinity and photocytoxicity. Besides, it could be demonstrated that EGF-HSA-SnCe6(ED) produced intracellularly ROS (reactive oxygen species) upon light irradiation, more than EGF-Dex-SnCe6(ED) did. It was concluded that, in contrast to EGF-Dex-SnCe6(ED) the photodynamic activity of the EGF-HSA conjugate of SnCe6(ED) on MDA-MB-468 breast adenocarcinoma cells is EGF-specific and more potent than free SnCe6(ED). PMID- 11004910 TI - [Treatment of IGA nephritis]. PMID- 11004911 TI - [Therapy options in systemic AL-amyloidosis with renal involvement]. AB - BACKGROUND AND OBJECTIVE: Despite significant efficacy of melphalan and prednisone in the therapy of systemic AL(light chain amyloid)amyloidosis the prognosis of the disease is poor. In patients with severe renal manifestation the reported results of low-dose melphalan therapy are inconclusive with respect to relief of clinical symptoms and overall prognosis. PATIENTS AND METHODS: We report our results of therapy in a group of 22 patients (8 women, 14 men, mean age 60 years) with renal involvement as the main manifestation of systemic AL amyloidosis without overt myeloma. RESULTS: Ten patients were treated with low doses of melphalan and prednisone. No significant clinical improvement was observed in any case: the patients died an average of 12 months after diagnosis of the disease. Three patients were treated with high doses of melphalan followed by autologous stem cell transplantation. One patient died due to septicaemia after high-dose chemotherapy. Two of the patients experienced significant remission and live virtually free of clinical symptoms 12 and 18 months after therapy. Nine patients were treated only symptomatically: four of them were alive an average of 30 months after diagnosis of systemic AL-amyloidosis. CONCLUSIONS: Only high-dose melphalan therapy offered a realistic chance of amelioration of clinical symptoms in our group of patients, although therapy-associated risks seem to be high. In patients with severe renal amyloidosis, who are not considered for high-dose therapy, particularly careful consideration of potential benefits and possible risks of conventional melphalan therapy is necessary, because the results of this approach are in doubt. PMID- 11004912 TI - [Hypocomplementemic urticarial vasculitis syndrome]. AB - HISTORY AND ADMISSION FINDINGS: A 40-year-old woman was admitted with recurrent membranoproliferative glomerulonephritis. For many years she had been suffering from joint pains and attacks of angio-oedema of the upper and lower lip. Physical examination was unremarkable except for marked oedema and urticaria of the trunk and limbs. INVESTIGATIONS: Skin biopsy revealed urticaria with vasculitis. Complement fractions C3, C4 and C1q were reduced, and C1q antibodies were demonstrated. DIAGNOSIS, TREATMENT AND COURSE: The findings of urticaria with vasculitis and reduced complement levels as cardinal symptoms together with joint pains, glomerulonephritis and C1q antibodies as minor symptoms were diagnostic of hypocomplementaemic vasculitic urticaria syndrome. The urticaria disappeared immediately when immunosuppressive treatment with prednisolone and cyclophosphamide was started, and the nephritic signs regressed. CONCLUSION: Differentiation of a vasculitic and a non-vasculitic form of chronic urticaria provides an initial step in delineating the underlying disease. Severe systemic disease such as collagen disease or vasculitis is not rare in vasculitic urticaria and requires long-term immunosuppression. PMID- 11004913 TI - [Early treatment of IgA nephritis is indicated by prognostic factors]. PMID- 11004914 TI - [Treatment of IgA nephropathies: a critical viewpoint]. PMID- 11004915 TI - [Vaccinations and medical recommendations for patients traveling after organ transplantation]. PMID- 11004916 TI - [Xenotransplantation and social risks]. PMID- 11004917 TI - [Renal artery stenosis]. PMID- 11004918 TI - [Nietzsche's medical legacy. An account on the centennial of his death]. PMID- 11004919 TI - [Emergency admissions of patients with stroke in the Munster area. A cross sectional study of the quality of regional health care]. PMID- 11004920 TI - [Delivery of hospital discharge reports to contract physicians. I]. PMID- 11004921 TI - [Delivery of hospital discharge reports to contract physicians. II]. PMID- 11004922 TI - [Publishing of negative results in experimental research]. PMID- 11004924 TI - Algorithms: let's give them back. PMID- 11004923 TI - [Human genome project: medicine in the light of evolution]. PMID- 11004925 TI - Irregular rhythm in a 67-year-old man. PMID- 11004926 TI - Comprehensive management of type 2 diabetes. PMID- 11004927 TI - Hysterectomy or an alternative? PMID- 11004928 TI - Chronic pain: 2. The case for opioids. PMID- 11004929 TI - A 55-year-old woman with a sore throat. PMID- 11004930 TI - Antimicrobial prophylaxis in surgery. PMID- 11004931 TI - Prevention of bacterial endocarditis. PMID- 11004932 TI - Medical education in the 'postgenomic era'. AB - The sequence of the human genome is very nearly in hand; the first draft has been completed, and the finished sequence will be available years ahead of schedule. Already, advances in medical genetics have affected the day-to-day practice of medicine by providing more powerful approaches to diagnosis of genetic disorders and cancer. But the full impact of the integration of genetics into medical practice lies before us. PMID- 11004933 TI - The dark side of the digital revolution. How to assess technology for potential side effects and contraindications. PMID- 11004934 TI - Menopause and mood. Is depression linked with hormone changes? AB - For most women, the perimenopause is not associated with depression. In those who do become depressed, mood symptoms tend to decrease at the end of menopause, regardless of previous history of depression. Many of the changes in brain function that affect mood are secondary to changing estrogen levels in the brain. Treatment with estrogen alone may improve mood in women with mild depressive symptoms, but those unresponsive to estrogen and women who have moderate to severe depression need antidepressant therapy. PMID- 11004935 TI - Weight gain during menopause. Is it inevitable or can it be prevented? AB - The years surrounding the menopause are associated with weight gain, increased central adiposity, and decreased physical activity. While weight change occurs independent of menopausal status, adverse changes in body fat distribution and body composition may be due to hormonal changes occurring during the menopausal transition. The one factor most consistently related to weight gain is physical activity. To avoid weight gain, women should make regular physical activity a priority. Although HRT use is widely believed to cause weight gain, data from the PEPI trial do not support this belief. Moreover, HRT may have a protective effect in reducing central adiposity, although more long-term studies using CT or MRI to measure visceral fat are needed to confirm this hypothesis. Data from the Women's Healthy Lifestyle Project provide clear evidence that weight gain and increased waist circumference, along with elevations in lipid levels and other CHD risk factors, are preventable through use of lifestyle intervention in healthy menopausal-aged women. Given the prevalence and chronic course of obesity, weight gain prevention should be recognized as an important health goal for women before they approach menopause. PMID- 11004936 TI - HRT to prevent cardiovascular disease. What studies show, how to advise patients. AB - How should you advise postmenopausal patients who are considering estrogen therapy? Do the potential cardiovascular benefits outweigh the risks? Does hormone therapy improve the cardiovascular risk profile in women with established coronary artery disease? In this article, Drs Wood and Cox help clarify the current clinical data for and against hormone replacement therapy for prevention of cardiovascular disease. In addition, they discuss current treatment recommendations and future directions in hormone therapy. PMID- 11004937 TI - Postmenopausal osteoporosis. Strategies for preventing bone loss, avoiding fracture. AB - Implementing osteoporosis prevention strategies through lifestyle modification and pharmacologic treatment reduces patient morbidity and mortality, as well as the cost to the healthcare system. All women should be advised about their risk factors and educated about options to reduce the risk. The primary care physician plays a crucial role in identifying women at risk who are most likely to benefit from intervention. Treatment decisions should be based on each woman's medical history and personal preferences. The optimal duration of treatment, especially with the newer agents, such as the SERMs and bisphosphonates, requires further investigation. PMID- 11004938 TI - Beta blockade in patients with congestive heart failure. Why, who, and how. AB - In patients with CHF, physicians should aim to treat the LV dysfunction, not just the CHF symptoms. LV dysfunction is a chronic disease that is usually progressive, even when it seem compensated. The risk of sudden death or progressive CHF is very real. Adding a beta blocker to the treatment regimen while the disease is still compensated or after resolution of an acute exacerbation can stabilize or reverse the LV dysfunction and improve survival. Beta blockade is now a vital part of the standard of care for most patients with LV dysfunction. PMID- 11004940 TI - Genetics beyond Mendel. Understanding nontraditional inheritance patterns. AB - Many medical conditions that clearly have a strong genetic component are not transmitted in a straightforward dominant, recessive, or X-linked pattern. Recent progress in understanding other modes of inheritance, such as imprinting, trinucleotide repeat expansion, mitochondrial inheritance, and mosaicism, has allowed us to solve many of these hereditary puzzles. Such advances have led to improvements in diagnosis and genetic counseling for patients affected with these disorders and should be valuable in development of effective therapies for some of these disorders in the future. PMID- 11004939 TI - Providing relief from headache pain. Current options for acute and prophylactic therapy. AB - The sometimes debilitating pain of migraine and other types of primary headache can be difficult to control with acute drug therapy alone. Often patients need both acute and prophylactic treatment to keep headache pain at bay. Dr Ward discusses the current treatment options for migraine with and without aura, tension-type headache, and cluster headache. He also provides practical pointers for use of the various formulations of specific drugs, including the newer "triptan" medications. PMID- 11004941 TI - Gallstones, from gallbladder to gut. Management options for diverse complications. AB - Gallstones may be incidental and asymptomatic or painful and accompanied by life threatening obstruction or infection. A thorough knowledge of potential complications is therefore critical, especially because some asymptomatic stones require prompt treatment. In this article, Drs Agrawal and Jonnalagadda provide valuable instructions for recognizing and treating the various manifestations of gallstone disease. PMID- 11004942 TI - The enlarging pearly nodule. Palisaded encapsulated neuroma. PMID- 11004943 TI - Menopause and mood. PMID- 11004944 TI - Darwin's beautiful notion: sexual selection and the plurality of moral codes. AB - One of the explicit objectives of Darwin's Descent of Man, and Selection in Relation to Sex was to explain cultural differences seen in human beings. Such an explanation, Darwin believed, was to rest upon an understanding of sexual selection. I examine the role that the beautiful plays within the mechanism of sexual selection as it works to differentiate isolated groups. It is suggested that an examination of the relationship between sexual selection and artificial selection--a relationship mediated by the beautiful--will illuminate key issues of the Descent as well as contemporary debates regarding the relationship between evolution and ethics. The beautiful, originally rooted in sexual selection, can become de-coupled from sexual selection and utilized in some selective process which is conscious. PMID- 11004945 TI - Darwin's species category realism. AB - Ever since Charles Darwin's On the Origin of Species was published, the received view has been that Darwin literally thought of species as not extra-mentally real. In 1969 Michael Ghiselin upset the received view by interpreting Darwin to mean that species taxa are indeed real but not the species category. In 1985 John Beatty took Ghiselin's thesis a step further by providing a strategy theory to explain why Darwin would say one thing (his repeated nominalistic definition of species) and do another (hold that species taxa are real). In the present paper I attempt to take this line of interpretation to a new level. Guided by the principle of charity, I provide and analyze a considerable amount of evidence from Darwin's mature writings (both private and published) to show that (contra Ghiselin and Beatty) Darwin did not simply accept the species delimitations of his fellow naturalists but actually employed, repeatedly and consistently, a species concept in a thoroughly modern sense, albeit with an implicit definition, a concept uniquely his own and fully in accord with his theory of evolution by natural selection. This implicit concept and definition is carefully reconstructed in the present paper. A new strategy theory is then provided to account for why Darwin would define species (both taxa and category) nominalistically on the one hand but delimit species realistically on the other. PMID- 11004946 TI - The evolution of complexity. Essay review. PMID- 11004947 TI - [Gastrointestinal metastases of lobular mammary carcinoma]. AB - Three women, two aged 63 and one aged 47 years, developed serious abdominal pains and nausea during and/or years after treatment for metastasized lobular mammary carcinoma. These symptoms were due to jejunal tumour with perforation, sigmoid and hepatic metastases and perforated gastric metastases, respectively. After surgical treatment of the affected part of the gastrointestinal tract, the patients survived for a number of months without abdominal symptoms. Infiltrating lobular mammary carcinoma is more often associated with gastrointestinal metastases than infiltrating ductal mammary carcinoma. Progressive abdominal symptoms attributable to such metastases constitute an indication for a change of the systemic treatment. In case of insufficient effect of this treatment, and in acute situations, surgical treatment may result in protracted palliation. PMID- 11004949 TI - [Immunology in the medical practice. XXXIII. Hereditary immune deficiencies: from genotype to phenotype]. AB - The advances of molecular biological techniques of the last decade have made possible identification of a rapidly increasing number of congenital (or primary) immune deficiencies at the genetic level. As soon as an immune deficiency is becoming a likely conclusion from a series of diagnostic laboratory tests, these recent developments in molecular techniques allow us in principle to make a conclusive diagnosis on the basis of a well-defined genetic defect. The hereditary immune deficiencies can be divided in 4 groups according to the functional immune aberration: antigen recognition, communication between immune cells, adhesion and directional motility, and killing as an effector mechanism. PMID- 11004948 TI - [Carrier state of x-linked adrenoleukodystrophy]. AB - X-linked adrenoleukodystrophy (X-ALD) is a sex-linked, inherited, metabolic disorder affecting the nervous system and endocrine organs. At least 20 to 50% of female carriers develop neurological deficits. Identification of female carriers is important, among other reasons because unnecessary new cases of this disorder, which is frequently lethal in boys, can be prevented by prenatal diagnosis. Furthermore, affected male offspring can be screened for adrenocortical insufficiency, which is treatable, or for early signs of cerebral involvement in which case bone marrow transplantation may be considered. Whether or not someone is a carrier can be investigated by determining the concentrations of saturated very-long-chain fatty acids in the plasma or cultured skin fibroblasts, by looking for the presence of X-ALD protein in cultured fibroblasts and by carrying out mutation analysis. Spasticity, painful muscular cramps, lumbago and arthralgias can be treated symptomatically with the same agents used for other aetiologies. A clinical geneticist can provide advice on heredity and the possibilities of prenatal diagnosis and pre-implantation techniques. PMID- 11004950 TI - [Pharmacotherapeutic compass 2000/2001]. AB - The Commission Pharmaceutical Help of the Health Care Insurance Board uses the Pharmacotherapeutical compass to inform the medical profession annually about medical drugs. In the 2000/2001 edition the most important alterations are changes in the introductions of: antidepressants, medicines for peptic disorders, medicines for rheumatoid arthritis and osteoarthritis, haematopoietic growth factors, anticonvulsants, medicines for Parkinson, medicines for congestive heart failure, antiretroviral medicines, medicines for osteoporosis, short acting insuline-analogues and oral antihyperglycaemics. The introductions about Horton's neuralgia and erectile dysfunction are new. For some (new) medicines recommendations are made, for the antidepressants and medicines for peptic disorders some recommendations have been changed. PMID- 11004951 TI - [Diagnostic image (2). (Gas gangrene of diabetic foot)]. AB - A 63-year-old woman with diabetes mellitus type 2 and an ulcer on the foot developed gas gangrene and was treated with antibiotics and surgical excision. She died before she could be treated with hyperbaric oxygen. PMID- 11004952 TI - [Increased proportion of elementary school children with asthmatic symptoms in the Netherlands, 1984/85-1994/95; a literature review]. AB - OBJECTIVE: To investigate whether the prevalence of asthmatic symptoms among children in the Netherlands has changed. DESIGN: Literature study. METHOD: Investigations into the prevalence of children with asthmatic symptoms were collected from Medline, Embase and various libraries if they had been performed between 1984/'85 and 1994/'95 and had used the so-called WHO or Region list. 23 studies were selected with data on 76,353 elementary school pupils. Linear regression analysis, weighed for the number of children, was used to determine if the proportions of children with asthmatic symptoms had increased over the years. RESULTS: In 10 years the prevalence of recent shortness of breath had increased by 107% (from 4.16 to 8.63%), that of recent wheeze by 16% (from 10.64 to 12.35%), that of recent attacks of breathlessness with wheezing by 17% (from 5.29 to 6.19%), and for chronic cough by 259 (from 1.05 to 3.77%) to 272% (from 3.14 to 11.68%). The prevalence of doctor-diagnosed asthma had increased by 120% (from 2.82 to 6.19%). CONCLUSION: There had been a pronounced increase in the percentage of primary schoolchildren with asthmatic symptoms between 1984/'85 and 1994/'95. The prevalence of doctor-diagnosed asthma increased faster then did key symptoms of asthma. PMID- 11004953 TI - [Relation between ethnicity and reason for referral of new patients to the Internal Medicine outpatient clinic of the Academic Hospital Rotterdam-Dijkzigt, 1997]. AB - OBJECTIVE: To inventory the ethnic composition of the patients referred to an internal medicine outpatient clinic of a Dutch academic hospital and to determine to what extent ethnic minorities differ from Dutch patients in terms of referral reasons, taking relevant background characteristics into account. DESIGN: Cross sectional analysis. METHOD: Data were collected on all new patients referred in 1997 for the first time to the internal medicine outpatient clinic of the Academic Hospital Dijkzigt, Rotterdam, the Netherlands, using the hospital information system (n = 3205). Patients were categorised into ethnic groups based on country of birth or name. Ethnic differences in referral reasons were tested for the 4 largest ethnic groups by means of logistic regression analysis with adjustment for age, sex, mean income of the zipcode area of the patients' residence and type of health insurance. RESULTS: The percentage of ethnic minorities amongst all referred patients was 22% (696/3205). The percentage of ethnic minorities among the patients referred from the catchment area of the outpatient clinic was 48% (209/440). Compared with Dutch patients Turkish patients were referred more often with stomach ache (odds ratio (OR): 4.26) and joint problems (OR: 7.16) as reasons. Moroccans were more often referred with stomach ache (OR: 4.10) and diabetes (OR: 4.51). Ethnic minorities were referred less often with dyslipidemia (Turks: OR: 0.11; Surinamese: OR: 0.17; Moroccans: 0 patients). PMID- 11004954 TI - [Cyanosis and cirrhosis of liver: hepatopulmonary syndrome]. AB - A 66-year-old man with Child-B alcoholic cirrhosis presented with melaena due to hemorrhagic gastritis. Clinical examination revealed cyanosis and clubbing accompanied by severe hypoxaemia without signs of obstructive or restrictive lung disease. Contrast-enhanced echocardiography showed right to left shunt due to intrapulmonary shunting leading to the diagnosis of hepatopulmonary syndrome. Hypoxaemia in patients suffering from chronic liver disease can have different causes but in the presence of cyanosis and clubbing without signs of lung disease the hepatopulmonary syndrome should be suspected. When presence of intrapulmonary shunts has been proven by contrast-enhanced echocardiography, a 100% oxygen test can determine the need for pulmonary angiography and thus determine the indication for interventional therapy or liver transplantation. The patient described was treated supportively with long-term oxygen therapy 6 l/min. Three months later his clinical condition was stable. An attempt to reduce the need for nasal oxygen failed. PMID- 11004955 TI - [Complementary therapies in the hospital:'if you can't beat them, join them'?]. AB - Some major cancer hospitals in the United States have established a department for complementary medicine as a service to their patients. Surveys in the Netherlands have shown that notably nursing staff and consumers consider the availability of complementary therapy in health care to be very important. Nevertheless, this appears to be an undesirable development as the effectiveness of these therapies has not been demonstrated and the therapists involved might interfere with regular patient therapy. PMID- 11004956 TI - [Publishing in biomedical journals in the year 2000 and after; report of a symposium]. AB - On the occasion of the oration by prof. dr. A.J.P.M. Overbeke, executive editor of the Nederlands Tijdschrift voor Geneeskunde, a symposium was organized on 16 June 2000. The lecturers were (chief) editors of medical journals. Many changes in medical publishing are to be expected in the next 10 years. The new technology will play an important part in this, but the details remain to be seen. Scientific journals can continue to exist in the electronic era if they produce legible, i.e. brief and clear articles in the printed version and offer extra information on the internet: details of methods used and statistical analyses and extensive tables. Also, the contributions of the various authors are printed. Not all, but only a few authors should make certain that the work described has indeed been performed and these would be responsible for the contents of the entire article. It appeared that the main part played by physicians in mass media is that of occasional informants. There still exist possibilities to use the media in distributing and supporting the messages of medicine and in elucidating the dilemmas. While 'archivist' journals mostly record scientific work presented, the task of the 'newspaper journals' consists of informing, interpreting, criticizing and stimulating. In this 'colouring' of the contents of the journal, the independence of the chief editors is of great importance. The innovations in medicine of the past century appear to be based mostly on the work of predecessors, of research physicians opposing established authorities, and on coincidence. The interval between discovery and publication has grown much shorter. PMID- 11004957 TI - [Familial hypercholesterolemia: recognition and prevention of cardiac complications at a young age]. PMID- 11004958 TI - Building teams, synergy, and your resources. PMID- 11004959 TI - Treating glaucoma. PMID- 11004960 TI - Deep brain stimulation for advanced Parkinson's disease. AB - Deep brain stimulation (DBS) is a new and promising technique for the treatment of movement disorders. Medically intractable Parkinson's disease (PD) is one of the most common indications for DBS. There are three possible subcortical targets for PD, depending on the symptomatology (i.e., the motor subdivision of the thalamus, the globus pallidus internus, the subthalamic nucleus [STN]). Thalamic stimulation has been well established as a safe and effective treatment for essential tremor and the tremor associated with PD. Globus pallidus internus and STN DBS are being investigated for the treatment of all the cardinal signs of PD. This article describes the pathophysiology of PD, the surgical treatment history of PD, surgical techniques used for DBS implants, and the role the perioperative nurse has in the care of the patients undergoing these procedures. PMID- 11004961 TI - Chronic regional pain syndrome, type 1: Part I. AB - Chronic regional pain syndrome refers to a class of disorders thought to involve common neuropathic and clinical features. These disorders usually are caused by injury, and they manifest in pain and sensory changes that are disproportionate in intensity, distribution, and duration to the underlying pathology. The result of these injuries is significant impairment of motor function over time. This article is divided into two parts. Part I discusses background information such as pain, pathophysiology, diagnosis, clinical stages, and the most common treatment modality, sympathetic nerve blocks. Part II, discusses alternate treatment modalities, such as sympathectomy, physical therapy, stimulators, trigger point injections, acupuncture, tourniquet effects, placebo effects, and amputation. PMID- 11004962 TI - Reducing percutaneous injuries in the OR by educational methods. AB - Exposure to bloodborne pathogens (e.g., HIV, hepatitis B, hepatitis C) through percutaneous injuries is an occupational risk for health care workers, especially those in the OR. The incidence of disease continues to rise, although occupational exposures often go unreported. Percutaneous injury prevention methods have included use of safety devices, practice changes, and educational programs. An educational intervention to increase awareness of risk, provide suggestions for injury reduction, and encourage reporting of exposures was performed at a university teaching hospital. Preliminary qualitative results show increased exposure reporting, increased use of personal protective equipment, and increased awareness of disease exposure risk among OR personnel. PMID- 11004963 TI - Perioperative nursing and animal-assisted therapy. AB - Interacting with animals has been shown to reduce blood pressure and cholesterol, decrease anxiety, and improve a person's sense of well being. Animal-assisted therapy (AAT) can be incorporated into the care of perioperative patients. Some of the goals that can be met by using trained and certified therapy animals are reducing stress preoperatively, motivating patients to have a positive attitude, promoting postoperative activity, and reducing the need for pain medication. Infection control, immunosuppressed patients, and research are issues regarding AAT in the hospital setting that must be covered by policy and procedure. Suggestions for setting up an AAT program are discussed. PMID- 11004964 TI - The art of fitting into our lives. AB - When there is a long-standing disparity between our reality and goals, internal friction grows unchecked. The incongruency of being out of step with ourselves creates major stress. Achieving congruency in our lives leads to personal and professional success. The process of achieving congruency brings us closer to our life goals, and it necessarily entails self-examination and insight. We must examine our likes, dislikes, strengths, weaknesses, and desires and set realistic and specific goals. By aligning our words and actions to match our goals, we are more likely to achieve those goals and begin fitting into the lives we want to lead. PMID- 11004965 TI - Storytelling can be a valuable teaching aid. PMID- 11004966 TI - [ELAN study]. PMID- 11004967 TI - [Characteristics of a cohort of 4,000 French patients with angina. The ELAN study]. AB - The ELAN (Etude Longitudinale dans l'ANgor) study was carried out to evaluate factors influencing the occurrence of death, myocardial infarction and revascularization procedures in patients with known angina pectoris. Analysis of baseline data collected in January 1997 involves 4,035 patients throughout France, which were recruited by 613 cardiologists practising on a private, hospital or mixed basis. The study population comprised 75% of men with a mean age of 65 years and 25% of women with a mean age of 70 years. Eighty eight percent of the patients had at least one cardiovascular risk factor, and nearly half of them had two or more factors; hypercholesterolemia and hypertension were the two most frequent ones. Reported cardiovascular past events included myocardial infarction in 47% of patients, PTCA in 33% and aorto-coronary bypass in 24%. Angina pectoris had been diagnosed within the previous year in 39% of patients. Exertional angina was the most common type (66%), with grade I/II angina being most frequently found (more than 70% of all cases). Management strategies are especially described for angina patients diagnosed within the previous year. More than half of the patients had undergone exercise testing within the previous 12 months, while scanning and coronary arteriography had been performed in 15% and 72%, respectively. Ninety five percent of patients were under antianginal drug therapy, with combined therapies being used in 58% of them. The most frequently prescribed drugs were betablockers (63%) and nitrates (53%). In 74% of patients, aspirin was given in addition to conventional antianginal agents. These data will be reviewed in a one-year cohort analysis as potential predictive factors for the occurrence of cardiovascular events. PMID- 11004968 TI - [One-year follow-up of a population of patients with angina. Factors influencing mortality and occurrence of cardiovascular events. Results of the ELAN study]. AB - The ELAN (Etude longitudinale dans l'angor) study was carried out both to acquire better knowledge of the occurrence of major cardiovascular events (myocardial infarction, revascularization surgery, death) in patients followed up for angina pectoris, and to determine the factors influencing such events. A cohort of angina patients was formed in January 1997, and 3,284 patients were followed up by 488 French cardiologists during a one-year period. Of these 3,284 patients, 96 (29/1000) died; causes of death included underlying coronary heart disease in 31, sudden death in 8, other cardiac aetiologies in 35, and noncardiac causes in 22. Sixty-nine (21/1000) patients developed myocardial infarction, 240 (73/1000) underwent PTCA, and 119 (36/1000) underwent coronary bypass surgery. Factors associated with an increased risk of death were age, diabetes mellitus, heart failure and angina type, mixed and rest angina being associated with poorer prognosis compared to exertional angina. Infarction risk increased with age and a history of previous infarction. Analysis of therapeutic factors after adjustment for the above risk factors showed a beneficial effect of betablockers on both cardiovascular and all-cause mortality and of aspirin on all-cause mortality. PMID- 11004969 TI - [Percutaneous angioplasty of unprotected left main coronary disease with implantation of systematic stenting. Immediate and mid-term results]. AB - The authors report the immediate and medium term results of percutaneous angioplasty of unprotected left main coronary disease with systematic stenting. Between March 1994 and December 1998, out of 6,006 patients undergoing coronary angioplasty, 92 had significant unprotected left main stem disease. The majority of patients was male (74 men, 80%) with an average age of 74.3 +/- 8.1 years. Between March 1994 and October 1996, only patients with a surgical contraindication were treated by angioplasty (n = 39). After October 1996, the indications were extended to patients who did not have surgical contraindications (n = 53). During the hospital phase, 4 patients (4%) died (ventricular arrhythmia: 1, cardiogenic shock: 2, gastro-intestinal haemorrhage: 1). No non fatal infarction with or without Q waves were observed, and no emergency coronary bypass surgical procedures were required. The angiographic success rate was 100%. During follow-up (7.3 +/- 5.8 months), 6 other patients died, 13 required a repeat coronary angioplasty, 4 for restenosis of the left main coronary artery, and 2 underwent coronary bypass surgery. The actuarial survival rate was 89 +/- 5% at 1 year and 85 +/- 17% at 3 years. Percutaneous angioplasty for unprotected left main coronary disease with systematic stenting was performed with acceptable hospital and medium term results. PMID- 11004970 TI - [Coronary stents in patients with severe left ventricular dysfunction. Hospital and long-term results]. AB - The aim of this study was to evaluate the risks and benefits, immediate and at long term, of coronary angioplasty associated with stent implantation in patients with severe left ventricular dysfunction. It was a retrospective study in which all patients with left ventricular ejection fractions 35% who underwent angioplasty between December 1994 and January 1998 were included. Seventy eight patients with an average ejection fraction of 29 +/- 6% who were haemodynamically stable were retained, excluding acute myocardial infarctions and cases of cardiogenic shock. The population was mainly masculine (6(men and 13 women) with a mean age of 65 +/- 11 years. The primary success rate was 97%. The loss of a collateral branch during the procedure, causing a non-Q wave infarction and the impossibility of implanting the stent at the desired site in another patient, were the only two failures. Hospital mortality was nil. The mean follow-up period was 450 +/- 290 days; long-term mortality was 17%. All deaths were of cardiovascular origin. The probability of survival at 6 months, 1 year and 800 days, was 88, 85 and 75% respectively. The good initial results were not maintained at long-term, but this could not be attributed to restenosis or to the pre-existing left ventricular dysfunction. PMID- 11004971 TI - [Cardiac abnormalities in a prospective series of 40 patients with type 2 diabetes]. AB - Cardiovascular mortality, the principal cause of early death in diabetics, is multifactorial. A prospective study was undertaken to analyse the different factors of excess cardiac complications in 40 patients with type 2 diabetes, whatever the symptomatology, by making an inventory of the cardiac abnormalities (systolic and diastolic left ventricular function, left ventricular hypertrophy, abnormalities of myocardial perfusion, heart rate variability and arrhythmias). Patients underwent 24 hour Holter monitoring, high amplification signal averaged electrocardiography, echocardiography, Thallium scintigraphy with a dipyridamole test followed by coronary angiography when positive. Patients were aged 60 +/- 8 years, diabetics for 11.8 +/- 6.8 years, and had associated cardiovascular risk factors: 85% were obese, 75% were hypertensive, 62.5% had hypercholesterolaemia and 60% were smokers. The HbA1C was 9.2 +/- 19%. An increased left ventricular mass was observed in 34.2% of patients. The left ventricular ejection fraction was normal (59.1 +/- 6.8%); 69.7% of patients had left ventricular diastolic dysfunction. Reduced heart rate variability was observed in 51.8% of cases. Late ventricular potentials were recorded on high amplification signal averaging in 39.5% of patients; 25.6% had significant ventricular extrasystoles and 52.2% had atrial extrasystoles. Twelve patients (45%) underwent Thallium myocardial scintigraphy with a positive dipyridamole test, 8 of whom had coronary lesions on angiography. The excess cardiac complications of diabetes is mainly due to ischaemic heart disease aggravated by autonomic neuropathy, left ventricular diastolic dysfunction, arrhythmias and left ventricular hypertrophy. In future, larger series are required to demonstrate that this detection can guide therapeutic intervention and reduce cardiac morbidity and mortality of diabetics. PMID- 11004972 TI - [Specific factors of diabetes in the rehabilitation of coronary patients]. AB - This prospective controlled trial included two groups of subjects with coronary artery disease: one of 33 patients with non-insulin-dependent diabetes mellitus (6 females and 27 males, mean age 57 +/- 9 years, left ventricular ejection fraction of 56 +/- 14%) and the control group of 33 subjects without diabetes (3 females and 33 males, mean age of 57 +/- 11 years, LVEF of 58 +/- 11%). Before training heart rate was faster for patients with diabetes at rest (72 +/- 11 vs 66 +/- 81 bpm, p = 0.03) and at the end of the stress test (127 +/- 15 versus 118 +/- 18 bpm, p = 0.03). After rehabilitation, resting heart rate and peak heart rate were similar. However cardiovascular capacities improvement was better in subjects without diabetes mellitus, especially concerning peak VO2 (28.98 +/- 8.88 versus 22.78 +/- 6.28 mL/min/kg, p < 0.01) and mechanical power (138 +/- 48 versus 118 +/- 23 watts, p < 0.01). Two groups were retrospectively distinguished among diabetic patients: one group of 17 patients showing a VO2 improvement superior to 5% and a second group non improved (VO2 < 5%). For the second group the two selective factors were higher fasting glycemia (1.83 +/- 0.75 versus 1.31 +/- 0.38 g/L, p = 0.01) and higher hemoglobin A1C (8.05 +/- 2.04 vs 6.62 +/- 1.03%, p = 0.02). Heart rate variability was not significantly different for these two groups: changes in autonomic nervous system cannot explain resistance of diabetic subjects to training. On the other hand, principal aim must be the equilibrium of glycemia in the management of diabetes. PMID- 11004973 TI - [Action of sotalol on high-amplification electrocardiogram]. AB - D,l sotalol is an antiarrhythmic widely used for treating ventricular excitability, especially ventricular tachycardia (VT). The means of assessing its efficacy is 24 hour Holter monitoring and programmed ventricular stimulation. High amplification ECG has also been proposed for predicting the effects of the drug on the induction of VT. The aim of this study was to assess the results of high amplification ECG before and after taking sotalol and to compare them with those of programmed ventricular stimulation. This study was performed in 24 patients with spontaneous and inductible ventricular tachycardia due to ischaemic heart disease. The two investigations were performed under basal conditions and after treatment with 160 to 320 mg/day of d,l-sotalol for 8 days. Nine patients had VT which could not be induced after treatment. In the other 15 cases, the VT remained inducible and was not significantly slowed. Analysis of different parameters of high amplification ECG, the QRS duration, RMS 40 and LAS did not show any difference after treatment in cases with inducible VT or VT suppressed by treatment. In conclusion, d,l sotalol, even at antiarrhythmic doses, does not seem to change the parameters of high amplification ECG, and it is therefore not necessary to stop treatment to carry out this investigation. Moreover, it was impossible to predict the effect of d,l sotalol on VT by high amplification ECG. PMID- 11004974 TI - [Use of echocardiography in the diagnosis of carcinoid tumors. Report of 4 cases]. AB - Carcinoid tumours are the most common neuro-endocrine tumours but cardiac involvement is rarely symptomatic although often observed at post-mortem and rarely revelatory of the disease. The authors report 4 cases in which echocardiographic detection of characteristic right ventricular involvement led to the confirmation of the diagnosis of carcinoid tumour leading to the secondary diagnosis of the primary carcinoid tumour. The clinical, physiopathological echocardiographic and therapeutic characteristics of this condition are discussed. PMID- 11004975 TI - [Paralysing hypokalemic thyrotoxicosis]. AB - Thyrotoxicosis is a rare cause of hypokalaemia. The clinical presentation associates signs of thyrotoxicosis which may be discrete, with severe hypokalaemia and peripheral muscular weakness with paralysis of the lower limbs. It is important not to miss this diagnosis because of the therapeutic implications. Treatment of this form of hypokalaemia is that of hyperthyroidism. PMID- 11004976 TI - [Chronic ischemic left ventricular dysfunction: myocardial hibernation?]. AB - Hibernating myocardium is a term which covers chronic ischaemic left ventricular dysfunction which is potentially reversible after revascularisation. Hibernating myocardium is classically associated with chronic hypoperfusion responsible for hypocontraction and cellular degeneration. This "classical" conception has been questioned as some workers emphasise that the reduction in coronary reserve responsible for repeated episodes of ischaemia and stunning could be the main causes of myocardial dysfunction. Position emission tomography (PET), and, most of all, myocardial scintigraphy and dobutamine echocardiography are the most commonly used techniques for detecting hibernating myocardium. Their sensitivity is good but the specificity and positive predictive value of dobutamine echocardiography seems to be better than the isotopic techniques. Structural abnormalities of hibernating myocardium and the delay, which is often long, between revascularisation and improvement, may explain some of the discordances between these techniques. Irrespective of the term used, hibernation or chronic ischaemic left ventricular dysfunction with myocardial viability, the reported data is in favour of coronary revascularisation with improved long-term quality of life and reduced mortality in patients with positive viability tests. PMID- 11004977 TI - [Tissue characterization by study of myocardial response to ultrasound. Review of the literature and perspectives]. AB - Echocardiography is a routine daily cardiological investigation. Recent technological developments suggest that it may become possible to quantify myocardial texture and thereby achieve histological and functional definitions of cardiac diseases. Two approaches are under evaluation: "videodensitometry" uses tools of statistical quantification from the echocardiographic image; more recently, analysis of the radiofrequency signal has been proposed. This is based on direct exploitation of a continuous signal at the transducer head of wide dynamic range containing all the information received by the piezo-electric crystals. These two approaches have given encouraging preliminary results in the recognition of different myocardial hypertrophic reactions, in diabetic cardiac disease and in the identification of myocardial viability. Despite the many remaining problems, the perspectives are promising and a new field of echocardiography should see the light of day. PMID- 11004978 TI - [Legionella myopericarditis]. AB - Cardiac involvement in legionella infection is rare but it is the most common extra-pulmonary complication. It usually takes the form of pericarditis, but a case of legionella myoparicarditis with global left ventricular hypokinesia on echocardiography has been described. The authors report a case of myopericarditis with massive pulmonary oedema and respiratory distress which regressed clinically and on echocardiography with reduction in chamber dilatation and complete recovery of left ventricular function. Legionellosis was confirmed on serology. The infection was probably contracted during a previous hospital admission, therefore, probably a nosocomial infection. Following the description of this case, a review of the literature is proposed. PMID- 11004979 TI - [Partial tricuspid homograft: a new technique for tricuspid valve repair. Report of a case]. AB - The authors report the case of a drug abuser treated successfully for fungal tricuspid endocarditis complicated by massive pulmonary embolism. Partial valvular replacement with a segment of a tricuspid homograft associated with disobliteration of a pulmonary artery was performed. Peroperative transoesophageal echocardiography showed satisfactory tricuspid valve function. The postoperative course was uneventful. After 5 months' follow-up, the patient was asymptomatic and in good general condition. Control echocardiography showed a stable operative result. PMID- 11004980 TI - [Valvular tumor: diagnostic trap. Report of a case of marastic endocarditis]. AB - Marastic endocarditis is a rare clinical condition described in cases of cancer or other severe inflammatory diseases. The authors report the case of a young patient in good general condition, admitted after a cerebro-vascular accident. Investigations showed an isolated mitral valvular mass on transoesophageal echocardiography which, after unsuccessful medical therapy, was operated. It was, in fact, a case of marastic endocarditis, and a pulmonary tumour was discovered one month after surgery. The bronchopulmonary adenocarcinoma had remained infraclinical beforehand. The advances in echocardiographic imaging will probably lead to an increase in such cases of early diagnosed thrombotic non-bacterial endocarditis (ETNB. This case suggests that it is justified to carry out an aetiological investigation of thrombotic non-bacterial endocarditis in all cases of isolated mitral valve masses. PMID- 11004981 TI - [Closure of the Aquitaine days of the French Society of Cardiology]. PMID- 11004982 TI - [Implications of latex allergy]. PMID- 11004983 TI - [Value of transthoracic aspiration puncture in the etiologic diagnosis of nosocomial pneumonia in patients not admitted to the ICU]. AB - The aim of this study was to determine the etiologic diagnosis of nosocomial pneumonia in hospitalized patients outside the intensive care unit. The study was carried out prospectively at the University Hospital Arnau de Vilanova in Lerida (Spain) from 1 May 1994 through 28 February 1996, during which 103 cases were diagnosed. Transthoracic needle aspiration (TNA) was chosen as the most useful diagnostic procedure, given its high specificity and low rate of complications. Sixty-seven TNA procedures were carried out; 10 (14.9%) were positive. Sputum was cultured for 62 patients and true positives were observed in 6 (9.7%). Pleural liquid was cultured for 35 patients and germs were isolated for 2 (5.7%). Bronchial brushing with a telescoping catheter was performed in 4 patients and brush culture provided 2 additional positives. Blood samples from 51 patients were cultured provided 4 additional diagnoses (7.8%). The most commonly found germs were P. aeruginosa (13.33%), S. aureus (13.3%), C. pneumoniae (10%) and L. pneumophila (10%). Disease was polymicrobial in 33.3%. Complications were observed, in the form of pneumothorax, in 2 cases (3%), one of which required drainage. We conclude that TNA, which is highly specific and has a low rate of complication, is a useful procedure for diagnosing nosocomial pneumonia in patients who are not being mechanically ventilated. PMID- 11004984 TI - [Assessment of excessive day-time sleepiness in professional drivers with suspected obstructive sleep apnea syndrome]. AB - Excessive daytime sleepiness is a common symptom of obstructive sleep apnea syndrome (OSAS) and can be a cause of traffic accidents, creating a problem of particular importance for professional drivers given the associated death, disability and professional repercussions. We assessed whether the Epworth sleepiness scale (ESS), which is a subjective measure of daytime sleepiness, correlates well with multiple sleep latency (MSL) testing, which gives an objective measure of daytime sleepiness. We also compared each method with the results of polysomnography (apnea-hypopnea index, arousal index and minimum oxygen saturation). We studied 55 professional drivers suspected of OSAS. All answered the ESS questionnaire and underwent polysomnographic and MSL testing. We found a significant, though not relevant, correlation between the degree of excessive daytime sleepiness estimated by the ESS and by MSL testing (r = -0.41; p = 0.002). A significant, though weak, correlation was found between the ESS score and the arousal index (r = 0.26; p < 0.05). Our results do not clarify which method is best for measuring excessive daytime sleepiness in professional drivers suspected of OSAS. PMID- 11004985 TI - [Usefulness of the IUATLD respiratory symptoms questionnaire for the differential diagnosis of bronchial asthma and chronic bronchitis]. AB - Our objective was to determine the diagnostic value of the questionnaire devised by the International Union against Tuberculosis and Lung Disease (IUATLD) for distinguishing between bronchial asthma and chronic bronchitis. We therefore compared clinical diagnoses established independently by two pneumologists for 211 patients to the patients' responses to the IUATLD questionnaire. The questions were analyzed for their ability to discriminate using the responses as independent variables and the diagnosis as the dependent variable. The individual predictive capacity of each question and the discriminating functions that identified the best clusters of questions were calculated using bayesian analysis. Finally, we compared IUATLD results to tests that assessed lung function (spirometry), obstruction variability (bronchodilator test, peak flow, bronchial challenge test), atopy (prick test, serum IgE), and clinical and biological markers (eosinophilia). The questionnaire correctly diagnosed 91% of the patients and a cluster of five questions registered a sensitivity of 85.6%, specificity of 91.4%, a positive predictive value of 93.1% and a negative predictive value of 82.2%, thus proving superior to the other tests. These results, along with the ease of administering the questionnaire allows us to consider the IUATLD instrument to be a good tool for the differential diagnosis of bronchial asthma and chronic bronchitis. PMID- 11004986 TI - [Morphometric assessment of rat lungs insufflated with liquid fixative at different pressures close to total lung capacity]. AB - INTRODUCTION: Liquid lung fixing through the trachea to a pressure of 25 cmH2O is currently accepted to be ideal. However, some studies do not seem to confirm that assumption. MATERIAL AND METHODS: The lungs of Fischer rats were filled with fixing liquid to four different pressures: 20 cm, 25 cm, 30 cm and 35 cmH2O. The fixed lungs were processed for inspection under a light microscope for morphometric study. The following variables were recorded: lung volume, tissue volume, air volume, internal alveolar surface (IAS), alveolar chord to measure the size of the distal air space, and the number of alveoli. Statistical comparisons were performed. RESULTS: Lung volume increased with insufflation pressure, with significant differences related to pressure increases from 20 cm to 25 cm and from 30 cm to 35 cmH2O. Air volume did not change, although tissue volume changed when pressure increased from 20 cm to 30 cmH2O and from 30 cm to 35 cmH2O. The increase of tissue volume was related to extravasation of interstitial fixer. The number of alveoli increased with pressure from 20 to 30 cm and from 30 to 35 cmH2O. IAS increased with pressure from 20 cm and all the other pressures. Alveolar chord, which is related to size of alveoli, decreased significantly as pressure increased from 20 cm to 25 cm. CONCLUSION: A pressure of 25 cmH2O is ideal for liquid fixing of lung volumes. With lower pressures the lung is partially distended and with higher pressures the fluid can pass into the interstitial space. PMID- 11004987 TI - [Assessment of mediastinal tumor involvement (T factor) in lung cancer with transesophageal ultrasonography]. AB - Transesophageal sonography was originally used to assess esophageal-gastric neoplasm. The technique may also be useful in the evaluation of pulmonary neoplasms for possible mediastinal involvement, with regard to both T and N factors. OBJECTIVE: To consider the validity of this minimally-invasive technique for assessing possible mediastinal involvement by direct tumoral invasion (T4) and to help obtain the most accurate staging of lung cancer. PATIENTS AND METHODS: Sixteen patients with confirmed histopathological diagnoses of pulmonary neoplasm were examined by transesophageal sonography in order to evaluate possible mediastinal involvement. They had previously been classified by computed tomography as T4 (12 patients) or possible T4 (4 patients). Mediastinal involvement was also assessed by mediastinotomy or thoracotomy in 15 of the 16 patients. RESULTS: Transesophageal sonography revealed mediastinal involvement in eight of the 16 patients; the rest had no such involvement. Surgical exploration of the mediastinum confirmed involvement in seven of those who had been so classified by sonography (with the remaining patient not having been assessed surgically). Among the eight patients who were considered free of mediastinal involvement, there was in fact none, although we found previously undetected infiltration of the posterior surface of the right pulmonary artery in one patient. Sensitivity was 87.5%, specificity 100% and accuracy 93.3%. CONCLUSION: Transesophageal ultrasound is a diagnostic tool that can provide additional information to complement other diagnostic strategies. PMID- 11004988 TI - [Rehabilitation of respiratory muscles in COPD]. PMID- 11004989 TI - [Beta-agonists in the treatment of bronchial asthma]. PMID- 11004990 TI - [Acute lipoid pneumonia caused by accidental aspiration of vaseline used in nasogastric intubation]. AB - Exogenous lipoid pneumonia is a rare disease which is often underdiagnosed. Exogenous lipoid pneumonia has traditionally been considered a chronic process secondary to continued aspiration or inhalation of fatty substances. In its acute form exogenous lipoid pneumonia sometimes coincides with accidental massive aspiration of lipidic material, as has been traditionally described for fire eaters. We review the literature and report a case of acute lipoid pneumonia secondary to accidental aspiration of vaseline used to place a nasogastric tube, which had been inserted for gastric lavage after a suicide attempt by flurazepam ingestion. PMID- 11004991 TI - [Hemothorax as complication of transjugular hepatic biopsy]. PMID- 11004992 TI - [Pleural effusion as presentation form of vertebral osteomyelitis of bacterial origin]. PMID- 11004993 TI - [Diagnosis of idiopathic pulmonary hemosiderosis]. PMID- 11004994 TI - Patient safety and clinical engineering. PMID- 11004995 TI - Patient safety and clinical engineering. PMID- 11004996 TI - Skin preparation to reduce ECG artifact. PMID- 11004997 TI - Successful PACS implementation: the importance of planning and maintenance. PMID- 11004998 TI - A feasibility study of multispectral image analysis of skin tumors. AB - To develop a noninvasive, early-detection method for skin cancers, a feasibility study of multispectral image analysis was investigated. The three most frequently occurring skin cancer types, ten basal-cell carcinomas (BCCs), ten squamous-cell carcinomas (SCCs) and five malignant melanomas (MMs) were studied, along with ten normal moles. Images were acquired by a charge-coupled device camera using eight narrow-band filters ranging from 450 nm to 800 nm, at 50-nm intervals. To extract main features of these tumors, principal components analysis (PCA) was performed, because it projects the multidimensional (here, eight-dimensional) data in the direction of maximum data variance. Then, the primary PCA components for red, green, and blue subset images were analyzed in terms of hue-saturation-intensity (HSI). By hue distributions, the BCCs and SCCs were differentiated from the MMs and normal moles. Texture information was used to further classify tumor types after the HSI analysis. The texture analysis, performed using a spatial gray level co-occurrence matrix (SGCM), could separate MMs from normal moles. The BCCs and SCCs were further studied by Fisher's linear discriminant analysis. Distribution was described as a Gaussian mixture model. By this classification procedure, seven BCCs, eight SCCs, five MMs, and ten NMs were correctly classified. Three BCCs and two SCCs were unseparable. Thus, multispectral skin cancer image analysis has potential to diagnose skin cancers. PMID- 11004999 TI - A novel device for measuring the effect of cholesterol on the release of oxygen from red blood cells into myocardial tissue. AB - A novel method has been devised to measure the effect of cholesterol on the release of oxygen (O2) from the red blood cell (RBC) into a tailored environment, which can be made to mimic myocardial tissue. Cholesterol affects the cell membrane of the RBC and thus the release of O2 into tissue. While this is true of all tissue, the myocardium is especially sensitive because of its critical nature, its high O2 requirements, and the shortness of time that arterial blood spends in the muscle. Calculations are presented that show that the release time for O2 from RBCs is close to the residence time of the RBC in the coronary system. Sequential measurements of blood oxygen saturation (SO2) are made when oxygenated blood is subjected to conditions similar to those in the myocardium. The natural logarithm of the relative value of the SO2 at time t compared with the initial value of the SO2 can be fitted to a straight line whose slope is proportional to the parameters of the RBC membrane, the sample size, the hematocrit, and the diffusion parameters of the apparatus. This value is used to estimate the effects of cholesterol-lowering treatments on O2 release. This test will serve as a valuable adjunct to or replacement for stress tests in the evaluation of coronary artery disease, especially in patients whose physical conditions make standard stress testing painful or risky. PMID- 11005000 TI - Implementing the FDA performance standard on electrode lead wires and patient cables in hospitals. AB - The U.S. Food and Drug Administration (FDA) Performance Standard on Electrode Lead Wires and Patient Cables became mandatory for all relevant devices on May 9, 2000. The standard requires that any lead wire or patient cable that has contact, temporary or permanent, with a patient, should not allow the connection of the patient to the earth or possibly hazardous voltages. This article advises those hospitals and other healthcare facilities that have not completed the upgrades of wires and cables on how to complete this task. PMID- 11005001 TI - Improving team performance: understanding the differences between teams and work groups. PMID- 11005002 TI - Network masks and Internet protocol addressing. PMID- 11005003 TI - Epidemiology of juvenile violence. AB - It is difficult to review the epidemiology of juvenile violence because few studies focus specifically on this topic as opposed to childhood aggression or delinquency in general. More research is needed specifically on juvenile violence, which is generally measured using official records or self-reports. Self-report research shows that a substantial fraction of the male juvenile population commits violence, and that very few violent acts are followed by arrests or convictions. Racial differences in violence may be explainable by reference to racial differences in community contexts. There is a great deal of versatility in juvenile violence. Juveniles who commit one type of violent offense also tend to commit other types and nonviolent offenses. Violent offenders tend to be persistent or frequent offenders, and there is little difference between violent offenders and nonviolent but equally frequent offenders. Nevertheless, there is some degree of specialization in violence. More research is needed to investigate whether risk factors exist for violence that are not risk factors for serious nonviolent delinquency (e.g., biologic factors). Violent juveniles tend to have co-occurring problems such as victimization, substance abuse, and school failure. Often, they might be described as multiple problem youth. There is considerable continuity from childhood aggression to juvenile violence. An early age of onset of violence predicts a large number of violent offenses. The major long-term risk factors for juvenile violence are individual (high impulsiveness and low intelligence, possibly linked to the executive functions of the brain), family (poor supervision, harsh discipline, child physical abuse, a violent parent, large family size, poverty, a broken family), peer delinquency, gang membership, urban residence, and living in a high crime neighborhood (characterized by gangs, guns, and drugs in the United States). More research is needed on interactions among risk factors, and especially on interactions between biologic and psychosocial risk factors. Important short-term situational factors include motives of potential offenders (e.g., anger, a desire to hurt), alcohol consumption, and actions leading to violent events (e.g., the escalation of a trivial altercation). More specific research is needed on protective factors against youth violence, for example, by investigating why aggressive children do not become violent juveniles. More research is also needed on the development and validation of risk assessment instruments. To investigate developmental and risk factors for juvenile violence, longitudinal studies are needed. Such studies should include multiple cohorts, to draw conclusions about different age groups, and should include both boys and girls and the major racial and ethnic groups. They should measure a wide range of risk and especially protective factors. They should be based on large, high-risk samples, especially in inner-city areas, incorporating screening methods to maximize the yield of violent offenders while simultaneously making it possible to draw conclusions about the total population. They should include long-term follow-up studies to permit conclusions about developmental pathways. They should make a special effort to study careers of violence and to link developmental and situational data. It will not be easy to mount new longitudinal studies focusing specifically on juvenile violence, but such studies are needed to advance knowledge about the epidemiology of juvenile violence, including risk factors and developmental pathways. PMID- 11005004 TI - Sociologic perspectives on juvenile violence. AB - In sum, there are four sets of social factors that help us understand why juvenile violence appears when, and where, it does, and why some communities and entire societies are persistently wracked by youth violence whereas others are largely spared its worst expressions. When it comes to the first three factors in particular--deprivation, disorganization, and brutalization--the evidence for these links is as strong as anything in social science, and that evidence is supported by a variety of sources and a variety of methods of investigation. Such investigation includes the knowledge we gain through social intervention. Some of the most effective violence prevention programs are successful precisely because they confront and deflect the social forces that otherwise often lead to violence. Consider, for example, the home-visiting programs that work with poor parents in disorganized communities to lower the risks of child abuse; and some of the more "holistic" or "multisystemic" efforts to work with violent juvenile offenders. The best of these programs work by tackling the problems of social isolation and lack of supports in the community, as well as immediate issues of economic survival for vulnerable families and children. More generally, we know that the availability of steady and rewarding work in the future, of the kind that can reliably sustain a family, is one of the most important factors allowing some youths to "desist" from violence as they mature. These conclusions give us much to be encouraged about, and much to be alarmed about. On the one hand, understanding that youth violence often is rooted in a set of adverse social conditions that are identifiable, and potentially modifiable, is a fundamentally optimistic message. It reminds us that the level of juvenile violence we suffer in America today is neither fated nor inevitable. Other societies that are in many respects much like us suffer far less of it; so could we, and we increasingly understand some of the ways in which we could make that happen. We know that some programs designed to increase opportunities and supports for high risk youth and families do work, and we know that there are broader social and economic policies--policies that are not abstract visions, but have actually been put into place in societies much like our own--that can significantly diminish the pressures toward violence that are bred by poverty and social insecurity among the young. But knowing what to do is one thing, and actually doing it is another. In this there is much reason for concern. The 7-year economic boom that began in the early 1990s has had salutary effects on youth violence, mainly by providing improved economic opportunities for low-income youth and, accordingly, diminishing the appeal of illegal activities. Despite that extraordinary burst of sustained prosperity, however, too many of America's youth remain impoverished, sometimes desperately so: the boom has had only limited effect on our disturbingly high rates of family poverty, which continue to tower above those of comparable industrial societies. And if the boom should come to an end, and we enter again into a period of rising joblessness for youth and young adults, then much, or all, of the recent gains could be quickly lost. The positive economic trends that have helped take the edge off serious juvenile violence in the past few years, in short, are both partial and fragile. Worse, some of our recent social and economic policies, at both state and federal levels, are working in the opposite direction. Some variants of welfare reform, for example, have tossed many vulnerable families off of public assistance without offering solid economic opportunities in its place. Many more families face this future as federal time limits on public welfare increasingly come into play. Given what we know about the links between deprivation and youth violence, this is not a development we can celebrate. (ABSTRACT TRUNCATED) PMID- 11005005 TI - Genetics and violence. AB - As is evident from this brief review, the genetic study of violence is maturing at an ever-increasing rate; much more work is needed to examine specific molecular genetic markers and their associated phenotypes, particularly in subjects with juvenile onset. It is also becoming clear that more work is needed to help delineate the specific phenotypes that are being transmitted within families. It is unlikely that we will find a gene or genes that are both necessary and sufficient for the manifestation of aggression and violence. It is more likely that specific genes will be associated with discrete factors that either increase the risk for the expression of violence or that are components of the violent phenotype. Thus, in addition to continuing research that examines the role of specific genetic factors in the manifestation of violence, more work is needed that will help further develop the nosology of violence and related behaviors. As noted previously, an understanding of the inherited phenotype is critical in the study of any disorder. It is especially important in the study of complex behaviors such as violence in which there may be several behavioral components that comprise the complete diagnostic category. PMID- 11005006 TI - Neurologic evaluation of violent juveniles. AB - Any juvenile who has acted violently requires a systematic, meticulous neurologic evaluation. There is no substitute for an adequate medical, family, and social history, the latter focusing on abuse and neglect. Evaluation should include a full physical and a conventional neurologic examination. Additionally, the physical, neurologic examination must include tests of cerebral-cortical function. The history and physical examination usually will indicate the diagnosis. A neuropsychologic evaluation of higher cerebral functions, including tests of frontal functions, should also be done in most cases. Some specific causes of brain disease can be identified by appropriate blood and urine tests, waking and sleep EEGs, and neuroimaging investigations. In general, positive findings are more significant than negative findings. If the physical examination and psychological tests are negative and the MR imaging shows a brain tumor, the patient has a brain tumor. If the neurologic or neuropsychological examinations demonstrate brain dysfunction, the patient has brain dysfunction, even if MR imaging is normal. PMID- 11005007 TI - Mental illness, neuropsychologic deficits, child abuse, and violence. AB - This review of the psychiatric, neuropsychological, and familial contributions to aggressive behavior makes clear that conduct disorder is not a single diagnostic entity. It is, rather, the final common pathway of the interaction among a variety of different kinds of intrinsic vulnerabilities and environmental stressors. In every aggressive child all of these vulnerabilities (none of which necessarily meets full criteria for a specific DSM-IV diagnosis) and stressors must be considered and, if present, addressed systematically. We know that psychotic symptomatology, especially paranoia, combined with neuropsychological vulnerabilities and a history of severe abuse become a recipe for violence, and the more impaired the child, if abused, the more violent the child will become as an adolescent and adult. The clinician must, therefore, think of himself or herself as the only knowledgeable adult who will ever take the time to discover these ingredients and deal with the violent child positively and therapeutically. PMID- 11005008 TI - Diagnostic evaluation of the violent child and adolescent. AB - The diagnostic evaluation of the violent child and adolescent is a collaborative exercise, requiring diverse expertise. Ironically, after systematic, comprehensive psychiatric, neurologic, psychologic, neuropsychologic, psychoeducational, family, and social evaluations have been completed, the clinician probably will not be able to make a hard and fast DSM-IV diagnosis; that is, unless the clinician is satisfied with fitting the behavioral pieces into the Conduct Disorder category and ignoring the rest of the clinical data. From time to time the clinician will be able to diagnose a bipolar mood disorder underlying a violent youngster's behavior. Occasionally the clinician will recognize paranoid schizophrenia motivating a particularly heinous or bizarre violent act. Most often the clinician will be faced with a variety of different kinds of psychiatric, neurologic, cognitive, and environmental vulnerabilities that have come together and created a violent child or adolescent. That's fine because almost invariably each discovered vulnerability has implications for treatment. The clinician evaluating a violent child or adolescent must overcome his or her initial disgust or anger at the youngster's behaviors and resist the impulse to dismiss the child as conduct disordered, as an incipient psychopath. That diagnosis leads nowhere. The identification of vulnerabilities leads to specific interventions and ultimately to the prevention of future violence. PMID- 11005009 TI - Treatments for aggressive and antisocial children. AB - Many different types of treatment have been applied to conduct-disordered youths. Unfortunately, little outcome evidence exists for most of the techniques. Three treatments with the strongest evidence to date were highlighted in this article: PMT, PSST, and MST. Parent management training is directed at altering parent child interactions in the home, particularly those interactions related to child rearing practices and coercive interchanges. Cognitive problem-solving skills training focuses on cognitive processes that underlie social behavior. Multisystemic therapy focuses on the individual, family, and extrafamilial systems and their interrelations as a way to reduce symptoms and to promote prosocial behavior. Each of these treatments has multiple controlled studies on its behalf, and some of the techniques (e.g., PMT) have been extraordinarily well evaluated. Significant issues remain to be addressed to accelerate treatment advances. We cannot yet say that one intervention can ameliorate conduct disorder and overcome the poor long-term prognosis. On the other hand, much can be said. Much of what is practiced in clinical settings is based on psychodynamically oriented treatment, general relationship counseling, generic family therapy, and group therapy (with antisocial youths as members). These and other procedures, alone and in various combinations in which they are often used, have not been evaluated carefully in controlled trials. Of course, absence of evidence is not tantamount to ineffectiveness. At the same time, promising treatments have advanced considerably, and a very special argument might be needed to administer treatments that have neither basic research on their conceptual underpinnings in relation to conduct disorder nor outcome evidence from controlled clinical trials on their behalf. Promising treatments, at best, leave important questions unanswered. Further development of treatments clearly is needed. Apart from treatment studies, further progress in understanding the nature of conduct disorder is likely to have very important implications for improving treatment outcome. Improved triage of patients to treatments that are likely to work will require understanding of characteristics of children, parents, and families that will make them more or less amenable to current treatments. PMID- 11005011 TI - Ethical implications of what we know about violence. AB - This article explores the moral implications of the previous articles in this volume. It looks at our knowledge regarding the neuropsychiatric and environmental influences on violence in light of concepts of justice, virtue, and responsibility. PMID- 11005012 TI - The centrosome and parthenogenesis. PMID- 11005010 TI - Forensic aspects of juvenile violence. AB - The juvenile justice system was created because it was recognized that youthful offenders needed to be managed differently from adults. They were to receive habilitation services instead of punishment. It is now more than a century since the creation of the first juvenile court. After 67 years, the US Supreme Court, in Kent v United States stated that the model was not working because juveniles in the criminal justice system received no treatment and they had no rights. Because the issue that had been appealed was the lack of rights (not lack of treatment), the Court mandated that juveniles, like adults, be given certain rights. The following year, in In re Gault, the Court expanded these rights. Subsequent Supreme Court cases have dealt with these kinds of issues--that is, whether juvenile offenders are entitled to the same rights as adults and subject to the same penalties. The Supreme Court has never heard a "right to treatment" case, which is the other part of the juvenile court system. Cases have been brought in lower courts (e.g., Nelson v. Heyne, 1972) alleging inadequate treatment services, but no national impact has resulted. Thus, in general, children in the juvenile court system do not have an enforceable right to treatment and can obtain only what services are available in their jurisdictions. The services often are woefully inadequate. Sentencing a youth to probation, with the requirement that he or she participate in counseling or mental health treatment, is meaningless if services are not available. Community-based, model programs that provide effective treatment do exist. They are, as yet, the rare exception rather than the norm and, therefore, are not available to most youthful offenders. Incarcerated juveniles, obviously, cannot avail themselves of community programs. Litigation to give these youth the same rights as adults in penal institutions is not the answer because incarcerated adults don't have a right to treatment, only a right to be free from "deliberate indifference" to their medical needs. It is hoped that a way will be found to provide effective services for the juvenile delinquent population. Federal laws have been enacted to mandate special services for educationally handicapped children. Those statutes, and litigation based on them, have led to some improvements for that group of children. Perhaps the same can be accomplished for the youthful offender population. Mandatory early intervention will serve them far better than mandatory waiver to adult court or incarceration in adult prison. PMID- 11005013 TI - gamma-Tubulin. PMID- 11005014 TI - gamma-Tubulin complexes and their role in microtubule nucleation. PMID- 11005015 TI - gamma-Tubulin of budding yeast. PMID- 11005016 TI - The spindle pole body of Saccharomyces cerevisiae: architecture and assembly of the core components. PMID- 11005017 TI - The microtubule organizing centers of Schizosaccharomyces pombe. PMID- 11005018 TI - Comparative structural, molecular, and functional aspects of the Dictyostelium discoideum centrosome. PMID- 11005019 TI - Are there nucleic acids in the centrosome? PMID- 11005021 TI - Centriole duplication and maturation in animal cells. PMID- 11005020 TI - Basal bodies and centrioles: their function and structure. PMID- 11005022 TI - Centrosome replication in somatic cells: the significance of G1 phase. AB - Proper cell division requires that the cell be able to form a bipolar spindle during mitosis. To achieve this, the centrosome must be replicated accurately during interphase. Our understanding of the mechanisms that allow centrosome doubling to be coordinated with other cell cycle progression processes is advancing at a rapid pace. Several different experimental systems have been developed that are allowing detailed studies of centrosome replication. For example, the identification of mutants in yeast that are unable to duplicate the SPB accurately during interphase has provided important insights concerning centrosome duplication. In addition, intact embryonic cells and extracts prepared from unfertilized eggs are powerful tools for investigating the molecular regulation of centrosome doubling during the cell cycle. Many of the observations from these embryonic systems are directly applicable to understanding centrosome doubling in somatic cells. Finally, transgenic mouse models and cultured mammalian cell systems have been developed for analyzing the regulation of centrosome doubling in cells with more complex cell cycles. As our knowledge of the cell cycle advances, particularly our understanding of the intricate series of events that must occur for somatic cells to traverse G1 phase, it should be possible to use the systems that have been developed to determine how the replication of the centrosome is coordinated with other cell cycle progression processes. The next few years should see rapid advances in our understanding of this critical cell biological process. PMID- 11005023 TI - The coordination of centrosome reproduction with nuclear events during the cell cycle. PMID- 11005024 TI - Regulating centrosomes by protein phosphorylation. PMID- 11005025 TI - The role of the centrosome in the development of malignant tumors. PMID- 11005026 TI - The centrosome-associated Aurora/Ipl-like kinase family. AB - Because of the well-known role of the centrosome and mitotic apparatus in genome partitioning in normal cells, defects in pathways essential for mitotic regulation are likely implicated in the cascade of events leading to aneuploidy and neoplasia. Exogenous overexpression of AIM-1, for example, produces multinuclearity in human cells and increased ploidy as well as aneuploidy (Tatsuka et al., 1998). Overexpression in colorectal tumor cell lines is thought to have a causal relationship with multinuclearity and increased ploidy. Cytokinesis error caused by AIM-1 overexpression is a major factor in the predisposition to cancer. As previously mentioned, the involvement of BTAK/aur2/AIK in centrosome amplification and its oncogenic activity are compelling. Aur2 has also been implicated in oncogenesis, and defects in kinetochore function leading to chromosome instability in human tumors should not be minimized (Farruggio et al., 1999). Further studies are needed to provide a clearer definition of how these kinetic proteins are linked and regulated in normal mitosis and cancer. Thus, Boveri appears to have been correct in formulating his early hypothesis that a defective mitotic apparatus and centrosome number were central and causative in chromosome missegregation and cancer. One hundred years later, at the onset of a new millennium and with light years of advanced technology in our favor, we are just now beginning to piece together the enzymes, substrates, and signaling pathways that support and explain his long-ignored but prophetic claim. PMID- 11005027 TI - Centrosome reduction during mammalian spermiogenesis. PMID- 11005028 TI - The centrosome of the early C. elegans embryo: inheritance, assembly, replication, and developmental roles. PMID- 11005029 TI - The centrosome in Drosophila oocyte development. AB - The Drosophila oocyte is a highly specialized cell type whose development utilizes MTOCs in various contexts. Figure 4 (see color insert) summarizes the characteristics of the MTOCs at different stages of oogenesis. Polarized mitoses are required to achieve oocyte determination. In the asymmetric germ-cell divisions that culminate in the egg chamber, the mitotic centrosomes are anchored to the spectrosome or fusome in order to produce the regular branching pattern of the cyst cells. It appears that the primary role of the fusome is to orchestrate the polarity and synchrony of oogenic mitoses. In the absence of fusomes or anchored spindles, the regular interconnected cyst network is lost and the oocyte does not differentiate. It is not known if the spindle itself is asymmetric, or whether either centrosome has equal potential to interact with the fusome. Several models can explain the need for polarized mitoses for oocyte differentiation. In one, an unequal distribution of unknown oocyte differentiation factors occurs from as early as the first cystoblast division. Here, the fusome may be required for the distribution of the factors. In another model, there is a mechanism that measures the number of ring canals in the cell, limiting the choice of oocyte to two potential pro-oocytes. In this model, polarized, synchronous divisions must occur to produce only two cells with the highest number of ring canals. In both of these models the centrosome plays an indirect role. A critical event in the determination of the oocyte is the formation of the MTOC. The oocyte MTOC forms shortly after completion of the germ cell mitoses and establishes a microtubule array along which factors required for oocyte determination are transported. It is unclear how this single MTOC forms in the 16-cell cyst, how the centrosomes become inactivated in the adjoining 15 nurse cells, or why the inactivated centrioles are transported into the oocyte. No molecular components of the MTOC are known except for centrosomin, which accumulates at the MTOC relatively late, at approximately stage 5 or 6 of oogenesis. The MTOC plays a central role in establishing the oocyte's polar coordinates. The oocyte microtubule array is required for the polar localization of axis-determining factors. At midoogenesis the MTOC appears to mediate the reversal of the microtubule array and the migration of the nucleus in the oocyte. The posterior follicle cells signal this reversal after receiving the gurken signal. What changes occur at the MTOC to trigger this cytoskeletal rearrangement? A better understanding of the MTOC's molecular components is necessary before we can begin to unravel the mechanisms underlying these events. The morphology of the MTOC changes after it shifts to the oocyte anterior. Staining with anti-centrosomin antibodies shows that the MTOC changes from discrete nucleus-associated bodies into a broad structure associated with the anterior cortex. The molecular mechanisms underlying this structural rearrangement of the MTOC at midoogenesis are presently unknown. Meiosis I occurs in the absence of centrosomes, but meiosis II spindles are linked by a shared, acentriolar, astral MTOC. The organization of the meiosis I spindle poles requires the NCD motor protein; however, the meiosis I spindle poles are acentriolar and contain no known centrosomal core proteins. The meiosis II astral spindle pole has a unique ring-shaped morphology and contains centrosomal proteins, such as gamma-tubulin. Strong mutations in the maternal gamma Tub37C gene do not block meiosis I, but prevent the progression of meiosis II. PMID- 11005030 TI - The centrosome in early Drosophila embryogenesis. PMID- 11005031 TI - Centrosome maturation. AB - In the past, centrosome maturation has been described as the change in microtubule nucleation potential that occurs as cells pass through specific phases of the cell cycle. It is suggested that the idea of centrosome maturation be expanded to include gain of functions that are not necessarily related to microtubule nucleation. Some of these functions could be transient and dependent on the temporary association of molecules with the centrosome as cells progress through the cell cycle. Thus, the centrosome may best be viewed as a site for mediating macromolecular interactions, perhaps as a central processing station within the cell. The centromatrix, a relatively stable lattice of polymers within the centrosome's PCM, could serve as a scaffold for the transient binding of mediator molecules, as well as allow the dynamic exchange of centrosome constituents with a soluble cytoplasmic pool. New evidence adds support to the idea that centrioles are crucial for the maintenance of PCM structure. However, significant evidence indicates that aspects of centrosome structure and function can be maintained in the absence of centrioles. In the case of paternal centrosome maturation, sperm centrioles may not contain an associated centromatrix. It is proposed that regulation of paternal centrioles or centriole associated proteins could mediate centriole-dependent centromatrix assembly following fertilization. Thus, regulation of centromatrix-centriole interactions could be involved in maintaining the integrity of the centrosome's PCM and play an important role in centrosome disassembly during cell differentiation and morphogenesis. PMID- 11005032 TI - Syndromes of disseminated intravascular coagulation in obstetrics, pregnancy, and gynecology. Objective criteria for diagnosis and management. AB - This article presents current understanding of the causes, pathophysiology, clinical, and laboratory diagnosis, and management of fulminant and low-grade DIC, as they apply to obstetric, pregnant, and gynecologic patients. General medical complications leading to DIC, which may often be seen in these patients, are also discussed. Considerable attention has been given to interrelationships within the hemostasis system. Only by clearly understanding these pathophysiologic interrelationships can the obstetrician/gynecologist appreciate the divergent and wide spectrum of often confusing clinical and laboratory findings in patients with DIC. Objective clinical and laboratory criteria for diagnosis of DIC have been outlined to eliminate unnecessary confusion and the need to make empiric decisions regarding the diagnosis. Particularly in the obstetric patient, if a condition is observed that is associated with DIC, or if any suspicion of DIC arises from either clinical or laboratory findings, it is imperative to monitor the patient carefully with clinical and laboratory tools to assess any progression to a catastrophic event. In most instances of DIC in obstetric patients, the disease can be ameliorated easily at early stages. Many therapeutic decisions are straightforward, particularly in obstetric and gynecologic patients. For more serious and complicated cases of DIC in these patients, however, efficacy and choices of therapy will remain unclear until more information is published regarding response rates and survival patterns. Also, therapy must be highly individualized according to the nature of DIC, patient's age, origin of DIC, site and severity of hemorrhage or thrombosis, and hemodynamic and other clinical parameters. Finally, many syndromes that are often categorized as organ-specific disorders and are sometimes identified as independent disease entities, such as AFE syndrome, HELLP syndrome, adult shock lung syndrome, eclampsia, and many others, either share common pathophysiology with DIC or are simply a form of DIC. These entities represent the varied modes of clinical expression of DIC and illustrate the diverse clinical and anatomic manifestations of this syndrome. PMID- 11005033 TI - Oral contraceptive pills and hormonal replacement therapy and thromboembolic disease. AB - The risk of thromboembolic complications with the use of second and third generation oral contraceptives is minimal and probably related to underlying congenital or acquired thrombophilic states. Estrogen dose-dependency leads to increased thrombin generation and increased plasmin generation. There is no convincing evidence that the balance between clotting and fibrinolysis is disturbed. The risk of venous thromboembolism with pregnancy is greater than with oral contraceptives. Hormone replacement therapy is safe for healthy women, and the benefits far outweigh the potential risks. PMID- 11005034 TI - Iron deficiency in pregnancy, obstetrics, and gynecology. AB - Iron deficiency remains a major health risk in the United States, despite the apparent availability of a high-quality diet. In the United States, at least 7.8 million adolescent girls and premenopausal women are iron-deficient. Worldwide, the challenge of identifying and treating iron deficiency is enormous. Physicians involved in the primary care and in the obstetric and gynecologic care of women of all ages must be aware of the nature of the problem and the correct approach to screening, diagnosis, and treatment. The potential benefit to newborns and infants and to their mothers is substantial. Furthermore, a thorough diagnostic evaluation has considerable potential for uncovering a potentially lethal disease, such as gastrointestinal malignancy, in a curable phase. PMID- 11005035 TI - Clinical and laboratory features and sequelae of deficiency of folic acid (folate) and vitamin B12 (cobalamin) in pregnancy and gynecology. AB - Classically, deficiency of folic acid (folate) or vitamin B12 (cobalamin) was recognized by the presence of a macrocytic anemia resulting from megaloblastic changes in the bone marrow. A markedly changing paradigm has identified both new mechanisms for altered folate and cobalamin status and new sequelae and clinical interrelationships that include altered mechanisms of absorption, a changing pattern of neurologic deficits, an increased risk of vascular occlusive lesions, and an important relationship with the mechanisms of neoplastic transformation. Several of these newer characterizations relate to issues of neoplasia in the nonpregnant woman and to issues in pregnancy, such as the potential for developmental abnormalities of the fetal nervous system. PMID- 11005036 TI - Gestational thrombocytopenia and immune thrombocytopenias in pregnancy. AB - Appropriate management of thrombocytopenia in the pregnant patient is important for the well-being of both mother and fetus. The healthy-appearing mother with mild thrombocytopenia may have either gestational benign thrombocytopenia, which does not produce fetal thrombocytopenia, or immune-mediated thrombocytopenia, which can produce fetal thrombocytopenia. These two types of pregnancy-associated thrombocytopenias can be differentiated. Gestational benign thrombocytopenia is initially discovered during pregnancy, and in these patients a reliable test for antiplatelet antibody is usually negative. Conversely, patients with immune mediated thrombocytopenia may have a history of thrombocytopenia before the pregnancy, and these patients usually have a detectable antiplatelet antibody. The pregnancy patient who presents with a normal platelet count and a history of neonatal alloimmune thrombocytopenia in a prior pregnancy or with a history of an infant of a close relative with NAT must be carefully monitored. Antiplatelet antibody assays performed on mother's and baby's blood will help determine if an antiplatelet antibody is present in maternal plasma, if the antibody reacts with the baby's platelets, and (with appropriate typing plasma) the antigenic specificity of the maternal and fetal platelets. In addition, antigenic typing of the father's platelets will help determine the risk of NAT in the current pregnancy. If a fetus is at risk for severe immune-mediated thrombocytopenia from either an autoantibody or an alloantibody, the fetal platelet count should be measured, if possible, from blood obtained by umbilical cord puncture. If the fetal platelet count is less than 50,000/microL or cannot be measured but is thought to have a high probability of being less than 50,000/microL, strong consideration should be given to a cesarean delivery. PMID- 11005037 TI - Recurrent miscarriage syndrome and infertility caused by blood coagulation protein or platelet defects. AB - Recurrent miscarriage syndrome and infertility are common problems in the United States. Recurrent miscarriage affects more than 500,000 women annually. If properly screened through a cost-effective protocol, the cause will be found in almost all women. The most common singular defect in women with RMS is a hemostasis defect, and if a thorough APLS evaluation is performed, the most common of these is found to be APLS. Other hereditary and acquired procoagulant defects are also commonly found, if looked for. It is important to evaluate women with RMS appropriately, because if a cause for the RMS is found, most women will achieve normal-term delivery. Hemorrhagic defects are rare hemostasis causes of RMS, but these defects also are treatable in many instances and should be considered in appropriate women. Treatment of the common procoagulant defects consists of preconception low-dose ASA at 81 mg/day followed by immediate postconception low-dose unfractionated porcine heparin. LMWH may be a suitable alternative. PMID- 11005038 TI - Antithrombotic therapy in high-risk pregnancy. AB - Venous thromboembolism remains a major cause of morbidity and mortality associated with pregnancy and puerperium. Specific risk factors for this disorder can be identified before or during pregnancy and delivery. The heritable defects believed to be associated with venous thrombosis are factor V Leiden mutation; elevated antiphospholipid antibodies; and deficiencies of antithrombin, protein C, and protein S. Women with a history of thromboembolism and thrombophilia should receive antenatal and postpartum thrombosis prophylaxis. PMID- 11005039 TI - Antithrombotic therapy in gynecologic surgery and gynecologic oncology. AB - The European Consensus Conference has assessed the risk for thrombotic complications for most women undergoing gynecologic surgery and found it to be moderate. Nonetheless, it is important to analyze a patient's individual risk before surgery so that appropriate thrombosis prophylaxis can be given if increased risk is determined. Malignancy accounts for most thrombotic complications among gynecologic patients. Patients with known malignancies should receive prophylaxis during surgery, and some patients with breast cancer should receive prophylaxis during chemotherapy. Heparin, and low-molecular-weight heparin in particular, may favorably influence the outcome of cancer in some patients and treatment with these agents is currently under investigation in a number of trials as a new approach to anticancer therapy. PMID- 11005040 TI - Hemorrhagic problems in obstetrics, exclusive of disseminated intravascular coagulation. AB - During pregnancy many physiologic changes occur that result in an increase in coagulation factors and a decrease in fibrinolytic activity. Because hemorrhage during pregnancy is a major cause of maternal morbidity, it is important to recognize and understand the pathophysiology of hereditary and acquired bleeding disorders. This article reviews von Willebrand's disease types 1, 2, and 3 and acquired hemophilia. PMID- 11005041 TI - Human platelet thrombin receptors. Roles in platelet activation. AB - Platelets are essential participants in hemostasis and thrombosis. Platelets normally circulate in blood as discoid resting cells that become critical constituents of hemostatic plugs or arterial thrombi only after specific receptors on platelet membranes interact with their ligands (agonists) to initiate the reactions that lead to platelet activation. The well-characterized events associated with platelet activation include activation of membrane receptors, shape change, granular secretion, cytoskeletal reassembly, platelet cohesion, and aggregation. The plasma protease alpha-thrombin is the most potent physiologic platelet agonist; this enzyme has other key roles in hemostasis, in the genesis of arterial thrombi, and in embryonic development, inflammation, wound healing, and cell proliferation. PMID- 11005042 TI - High-dose right unilateral ECT. PMID- 11005043 TI - Quantitative EEG during seizures induced by electroconvulsive therapy: relations to treatment modality and clinical features. I. Global analyses. AB - This study examined the effects of electroconvulsive therapy (ECT) treatment conditions, patient individual difference factors, and clinical outcome on global electroencephalogram (EEG) power during and immediately following ECT-induced seizures. Sixty-two patients were randomized to ECT conditions differing in electrode placement (right unilateral versus bilateral) and stimulus dosage (just above seizure threshold versus 2.5 times seizure threshold). At the second and penultimate treatments, global total power (1.5-28.5 Hz) and global power in specific frequency bands were quantified in 19-lead EEG recordings of the generalized seizure and the immediate postictal period. Seizures induced with high dosage, and to lesser extent, with bilateral electrode placement, resulted in greater global power. Patient age, initial seizure threshold, and baseline depression severity were inversely related to global power during seizures. While superior clinical outcome following ECT was associated with greater global power during seizures, this effect was small. The factors associated with more robust seizure expression also resulted in greater postictal bioelectric suppression. Associations with treatment parameters and patient variables were stronger at the second than penultimate treatment. We conclude that manipulations of ECT technique strongly determine the magnitude of seizure expression, but relations with clinical outcome are weak. The findings raise doubt about the clinical utility of algorithms based on analysis of EEG features to guide ECT parameter selection. PMID- 11005044 TI - Quantitative EEG during seizures induced by electroconvulsive therapy: relations to treatment modality and clinical features. II. Topographic analyses. AB - This study tested three alternative theories of the mechanisms of therapeutic action of electroconvulsive therapy (ECT). The theories differed in predictions about the global and topographic effects of effective and ineffective forms of ECT on electroencephalogram (EEG) seizure expression. At the second treatment, 19 lead EEG recordings were obtained in 57 depressed patients randomized to conditions that differed in ECT electrode placement and stimulus dosage. Power in the delta frequency band was quantified during the seizure and analyzed with traditional multivariate methods and the Scaled Subprofile Model. Electrical dosage of the ECT stimulus had a powerful effect on ictal global delta power and, more so, than electrode placement. Greater ictal global delta power was associated with superior therapeutic outcome, but the magnitude of this effect was small. Effective forms of ECT resulted in a topography where delta power was accentuated in prefrontal EEG sites. High dosage right unilateral ECT also resulted in stronger asymmetry in prefrontal regions than the ineffective, low dosage right unilateral ECT. Greater bilateral generalization of seizure expression does not appear to be a prerequisite for therapeutic effects. Instead, more intense seizure expression in prefrontal regions may be critical for efficacy. PMID- 11005045 TI - The effect of repeated bilateral electroconvulsive therapy on seizure threshold. AB - Seizure threshold was measured by empirical titration in 28 patients referred for bilateral electroconvulsive therapy to treat depressive illness at the outset of treatment and after another six treatments. No patient was given antiepileptic drug treatment, and anesthetic technique and concomitant psychotropic drug treatment were fixed. The average (+/- SD) initial seizure threshold measured by set charge was 79.5 mC (+/- 33.4 mC), and this increased to 95.5 mC (+/- 37.9 mC). The average percentage increase was 22.8% (95% confidence interval, 13.7% to 31.8%). The seizure threshold measured by set charge did not change in 15 patients (54%), and there was no significant relation between change in seizure threshold and patient sex, change in seizure duration measured by cuff technique, or global clinical improvement during the course of treatment. PMID- 11005046 TI - Electroconvulsive therapy and falls in the elderly. AB - Electroconvulsive therapy (ECT) is commonly used in the practice of geriatric psychiatry. Although it has been found to be both safe and effective, one of the most common complications associated with ECT is falls. A logistic regression model was created to identify independent risk factors for falls in the elderly. Two risk factors were identified: the number of ECT treatments and the diagnosis of Parkinson disease. Further studies are needed to develop strategies aimed at reducing the number of falls. In the meantime, clinicians need to be aware that falls represent an important complication associated with ECT. PMID- 11005047 TI - Seizure threshold estimation by formula method: a prospective study in unilateral ECT. AB - Formula methods of estimating seizure threshold in bilateral electroconvulsive therapy (ECT) have been successful in 75% (at the first ECT) and 80% (at the sixth ECT) of treatments (Gangadhar et al., 1998). This study showed the same results for unilateral (UL) ECT patients. Its aim was to compare formula and titration methods for threshold determination. The seizure threshold (dependent variable) was determined by the titration method used at the first ECT in consecutive consenting patients (n = 80) prescribed UL ECT under general anesthesia. The independent variables were age, gender, diagnosis, illness severity, concurrent drugs, head circumference, and inion-nasion distance. Forward, step-wise, linear regression analysis showed age as the only significant predictor of seizure threshold (15% of variance). A formula based on regression analysis was prospectively applied in an independent sample (n = 30) of patients receiving UL ECT using the titration method for threshold determination. The results calculated a higher threshold than the actual threshold used in 14 patients, a threshold level in 8 patients, and below threshold in 8 patients. Formula-based estimates would have been successful in 22 (73%) patients, but the majority of them would have received higher than the recommended stimulus dose. Titration is the method preferred for clinical use. However, if a patient's doctor wishes to use the formula-based method, he or she should do so with specific considerations. PMID- 11005048 TI - Post-ECT agitation and plasma lactate concentrations. AB - This prospective study evaluated the hypothesis that emergence agitation after electroconvulsive therapy (ECT) could be caused by lactate-induced panic secondary to insufficient neuromuscular blockade. Plasma lactate levels were measured before and after 245 consecutive ECT sessions in 37 patients monitored for evidence of post-ECT agitation. ECT was administered using a brief-pulse, rectangular, constant-current device through bilaterally placed electrodes under general anesthesia and neuromuscular blockade. Agitation was observed in 7% of all ECT sessions. No significant difference could be found in pre-ECT lactate levels. However, mean post-ECT lactate levels in agitated sessions were significantly greater than those in nonagitated sessions (4.77 versus 2.54 mmol/l, p < 0.05). An increase (+27%) in the pre-ECT succinylcholine dose for those patients who previously had repeated post-ECT agitation resulted in cessation of post-ECT agitation and return of the formerly high post-ECT lactate levels to normal (1.61 versus 2.07 mmol/l). Although the number of patients who had post-ECT agitation was small, the data support the hypothesis that post-ECT agitation might be a manifestation of lactate-induced panic. PMID- 11005049 TI - Electroconvulsive therapy and the alpha-2 noradrenergic receptor: implications of treatment schedule effects. AB - Six factorially designed studies evaluated the effects of different schedules of electroconvulsive shocks (ECSs) on alpha-2 adrenoceptor function in the rat brain. Attenuation of the hypomotility response to a clonidine challenge was taken to indicate alpha-2 adrenoceptor downregulation, a putative mediator of antidepressant action. Six daily and six alternate-day ECSs were shown separately to produce this receptor change. Three alternate-day ECSs produced comparable downregulation for a comparable period as six daily ECSs; this suggests that ECS produces time-dependent effects. No changes were elicited with a single ECS, which indicates that a single ECS may not influence alpha-2 receptor function. Three daily ECSs produced brief downregulation, which has implications for receptor dynamics as a function of the ECS schedule. Finally, maintenance ECSs sustained alpha-2 adrenoceptor downregulation over 6 weeks, which suggests a possible neurochemical basis for maintenance electroconvulsive therapy. The clinical relevance and scope for further research are discussed. PMID- 11005050 TI - A statewide survey of ECT policies and procedures. AB - The 1990 American Psychiatric Association (APA) Electroconvulsive Therapy (ECT) Task Force Recommendations include facility policy and procedure guidelines. The objectives of this study were to determine and to improve the adherence to the 1990 APA ECT Task Force Recommendations on policies and procedures among the providers of ECT in Louisiana. Completed surveys on ECT policy and procedures were obtained from the seven major Louisiana ECT providers from the last quarter of 1996. Project coordinators distributed copies of the survey results and a comprehensive set of ECT policies and procedures at a statewide meeting of participating hospitals during the spring of 1997. Most facilities had policies for electrical safety of ECT equipment, testing of new ECT equipment, pre-ECT work-up, ECT informed consent, patient instruction sheets, outpatient ECT, documentation of ECT procedures, clinical privileging, and ECT quality assurance monitoring. Subsequent telephone follow-up found that all participants changed their policies and procedures as a result of the project. Louisiana ECT providers showed general compliance with the facility policy and procedure aspects of the 1990 APA ECT Task Force Recommendations. The awareness model of guideline compliance was applicable to improving facility policies and procedures. PMID- 11005051 TI - ECT in a patient with a deep brain-stimulating electrode in place. AB - A 68-year-old woman had a deep brain-stimulating electrode placed for worsening of a long-standing essential tremor. There is currently no data available on the use of electroconvulsive therapy (ECT) in the presence of the deep brain stimulator (DBS). We report here the successful use of ECT in this patient with no adverse effects to the patient or to the DBS. No special provisions were made with the ECT administration except to ensure that the stimulator was turned off prior to administration of the ECT series, and remained off through the eight treatments administered to this patient. Following the course of treatment, her mood was markedly improved, and she displayed only minor postictal confusion. PMID- 11005052 TI - ECT in a patient with psychotic retarded depression, metastatic hepatic cancer, and esophageal varices. AB - We report a patient with liver disease and esophageal varices who responded to intensive psychiatric intervention on a medical unit. Her case illustrates the successful use of electroconvulsive therapy in the presence of varices, and highlights some precautions that could be taken in treating similar patients. PMID- 11005053 TI - ECT administration in a patient after craniotomy and gamma knife surgery: a case report and review. AB - The safe administration of electroconvulsive therapy (ECT) in a man with steroid induced depression and a history of craniotomy and gamma knife surgery for two separate foci of metastatic laryngeal cancer is reported. This is the first reported case of ECT given to a patient with a history of gamma knife surgery. The literature on ECT administration to patients with brain masses or a history of craniotomy is reviewed. The current case report indicates that with careful medical evaluation, including consultation with neurosurgeons and internal medicine physicians, ECT may be safely administered to this patient population. PMID- 11005054 TI - Lithium and maintenance ECT. AB - A 78-year-old man with a long history of major depression responded well to a course of ECT but relapsed before maintenance treatments could be started. Addition of lithium carbonate resulted in a sustained remission with maintenance treatments. The combination of maintenance ECT and lithium may be safe and effective in selected cases of refractory depression. PMID- 11005055 TI - Severe self-injurious behavior associated with treatment-resistant schizophrenia: treatment with maintenance electroconvulsive therapy. AB - A 35-year-old man with schizophrenia associated with a 10-year history of repetitive and often severe self-injurious behaviors was treated successfully with maintenance electroconvulsive therapy. Initially his condition did not respond to multiple psychotropic agents (including a year-long trial of clozapine), 27 hospitalizations, two courses of bilateral electroconvulsive therapy, and supportive psychotherapy with partial hospitalization. After the initiation of maintenance electroconvulsive therapy, this patient was able to tolerate residential placement and begin a work therapy program with only one instance of self-injurious behavior in the past 17 months. PMID- 11005056 TI - Ventricular ectopy associated with low-dose intravenous haloperidol and electroconvulsive therapy. PMID- 11005057 TI - The rate of ECT use in young people. PMID- 11005058 TI - Certification in ECT: a rush to judgment. PMID- 11005059 TI - Transient dissociative identity disorder after electroconvulsive therapy. PMID- 11005060 TI - Effectiveness of ECT against delirium during an episode of bipolar disorder: a case report. PMID- 11005061 TI - The map of dying. PMID- 11005062 TI - Infectious diseases and medical microbiology: a new training programme. PMID- 11005063 TI - The control of pain in palliative care. PMID- 11005064 TI - The ethics of clinical research in developing countries. PMID- 11005065 TI - The Academy of Medical Sciences. PMID- 11005066 TI - Unravelling the heterogeneity of non insulin dependent diabetes. PMID- 11005067 TI - Proteinuria in diabetes. PMID- 11005068 TI - Clinical management of the painful diabetic neuropathies. PMID- 11005069 TI - Diabetes in African Caribbean, and Indo-Asian ethnic minority people. PMID- 11005070 TI - Principles of pain control in palliative care for adults. Guidance prepared by a Working Group of the Ethical Issues in Medicine Committee of the Royal College of Physicians. AB - Specialist palliative care is now widely available and there are many good textbooks of palliative medicine. But some patients still suffer pain needlessly. Clinicians may not know how to use analgesia appropriately or may be anxious about giving an adequate dose of strong opioids such as morphine. This brief outline is intended to make the principles of pain control readily available to all clinicians who look after terminally ill patients. It applies to patients with non-malignant disease as well as to those with advanced cancer. PMID- 11005071 TI - Discriminating for the ageing population--the positive approach. PMID- 11005072 TI - Towards an improved career structure for non-consultant career grade doctors. PMID- 11005073 TI - Imaging in medicine through the 20th century. PMID- 11005074 TI - A new joint training programme in infectious diseases and medical microbiology. AB - The increasing overlap between the disciplines of medical microbiology and infectious diseases prompted the Joint Royal Colleges Committee on Infection and Tropical Medicine to set up a working party to examine how trainees could obtain certification in both subjects. Following widespread consultations, a scheme was developed that entails six years of training and leads to the award of CCSTs in both microbiology and infectious diseases. Both Royal Colleges and the Specialist Training Authority have approved the scheme. Joint training will be demanding and will not be suitable for everyone; it represents an alternative approach to training in the infection disciplines that will run alongside the existing monospecialty training programmes. PMID- 11005075 TI - The potential role of IT in supporting the work of junior doctors. AB - OBJECTIVES: To develop an information system using hand-held personal computers to support the work of junior doctors. DESIGN: A user-driven design process was used. Functionality reflected the core work of house officers (HOs) and senior house officers (SHOs); i.e. ward round lists, immediate discharge summaries, patient handover, laboratory and guidelines reference information and a personal record of clinical experience. SETTING: Salford Royal Hospital NHS Trust is a university hospital with a large acute medical case load. SUBJECTS: Several 'generations' of HOs and SHOs working in two of the four medical firms. Predominantly acute medical case load. MAIN OUTCOME MEASURES: Robustness, acceptability and benefits, assessed by observations and questionnaires. RESULTS: The system has proved robust. It is time neutral for the juniors and minimal time is needed for training/adaptation. Most felt the system to have either made their lives easier or to have had no detrimental impact. Ward round lists and printed immediate discharge summaries have been particularly successful. CONCLUSIONS: A hand-held IT system designed for house doctors can provide immediate, low-cost support for their activities. PMID- 11005076 TI - Coping with pressures in acute medicine--the second RCP consultant questionnaire survey. AB - The second questionnaire survey of consultant physicians involved in acute unselected takes in 1999 achieved a 76% response rate, and the results have been compared with those from the first survey of 1997. The proportion of consultants whose trainees worked partial shifts had increased from 42% to 61%, although these patterns of duty were adjudged to have detrimental effects on the quality and continuity of care, and on junior staff education and training. The benefits of ward-based systems were counterbalanced by their disadvantages, but introductions of admission wards and assessment units were considered a resounding success. The number of hospitals with 'physician of the week' schemes had increased from 12 to 23, but opinion of their value was sharply divided. The provision and competence of all grades of locums was identified as an increasing problem. Seventy per cent of respondents stated that they would never participate in 'hands-on' emergency care, although 86% thought that future consultants might have to do so. Seventy-nine per cent reported increases in the pressures of their posts and in their working hours, and the tensions between general and specialist duties were highlighted. Most consultants considered that the only long-term solution to the staffing crisis was a marked expansion in the numbers of all grades of medical staff. PMID- 11005077 TI - Controversy in thyroid disease. AB - Effective management for hypothyroidism has been available for a century, thanks to the pioneering efforts of George Murray, who first tested injection of thyroid gland extracts in Newcastle. Radioiodine for the diagnosis and then treatment of Graves' hyperthyroidism was introduced by Hertz and Roberts in Boston, and by Leblond in Paris, in 1943, the same year that Astwood in the USA began to use the predecessors of modern antithyroid drugs. Despite this lengthy history, debate continues as to the most effective management of these common disorders. More use of radioiodine, including its use in euthyroid goitre, is being advocated as its safety is now well established. Recent developments in optimising these treatments will be considered in this brief review. PMID- 11005078 TI - Virus hepatitis update. AB - Currently seven viruses, A, B, C, D, E, G and transfusion transmitted virus (TTV), are recognised in the hepatitis virus alphabet. Hepatitis G virus and TTV probably do not cause liver disease in humans. Hepatitis A and E usually cause a self-limiting hepatitis followed by complete recovery but occasionally cause fulminant hepatic failure. Hepatitis B and C are major public health problems worldwide due to their sequelae of chronic hepatitis, cirrhosis and primary liver cancer. Chronic hepatitis C is a particular health issue for Western Europe already, accounting for 40% of end-stage cirrhosis and 30% of liver transplants. The contribution of hepatitis C to chronic liver disease is predicted to rise in the future. Vaccines can prevent hepatitis A and B. Interferon alpha is effective treatment in 25-30% of patients with chronic hepatitis B or C. The prospects for treating chronic hepatitis B have been improved by the introduction of reverse transcriptase inhibitors. Lamivudine is the first drug of this class to be licensed. The optimal use of these new drugs is currently being studied. The success rate for treating chronic hepatitis C can be raised to about 40% with combination therapy of interferon alpha and ribavirin. A large research effort to discover new antiviral agents against hepatitis C is already giving the prospect of more effective therapies in the next few years. PMID- 11005079 TI - Medicine in the millennium: confronting the issues. AB - This joint conference of the Royal College of Physicians of London, the Royal College of Surgeons of England and the Royal Society of Medicine took place on 1 November 1999 at the Royal Society of Medicine in London. PMID- 11005080 TI - A NICE winter conference. PMID- 11005081 TI - Insurance medicine. PMID- 11005082 TI - The consultation in art. PMID- 11005083 TI - Balzac and cretinism. PMID- 11005084 TI - Alcohol abuse and the burden on the NHS. PMID- 11005085 TI - The National Clinical Guidelines for Stroke. PMID- 11005086 TI - Withdrawing and withholding life-prolonging medical treatment. PMID- 11005087 TI - Health care cost in Africa. PMID- 11005088 TI - [Surgery of the external auditory canal in familial bilateral auditory canal atresia]. AB - BACKGROUND: Inherited isolated bilateral atresia of the external auditory canal is rare. Ear canal surgery procedure is difficult. METHODS: Basing on a case report of a family with inherited isolated bilateral atresia of the external auditory canal in 4 cases the way of preoperative diagnostics including human genetics and our surgical concept is described. RESULTS: Resulting clinical findings showed complete epithelialization of the ear canal and ear drum with slight conductive hearing loss. CONCLUSIONS: Canaloplasty in atresia can be easily and successfully accomplished by our modified technique. PMID- 11005089 TI - [Prediction of verbal and numerical comprehension in patients with cochlear implants]. AB - BACKGROUND: Cognitive abilities determine the outcome of cochlear implantation. Neuropsychological tests of intellectual, memory, attentional and emotional functions were applied pre-operatively in 33 deaf patients receiving a cochlear implant. The outcome of verbal and numerical comprehension was measured in a subgroup of 14 patients post-operatively and correlated with pre-operative neuropsychological performance. METHODS: Neuropsychological performance was recorded using standardized tests for intellectual abilities (abbreviated Wechsler Adult Intelligence Scale [WAIS], crystallized intelligence scale), memory performance (Benton Visual Retention Test), attentional functions (d2 letter-cancellation-test), reaction time (Vienna Reaction Test) and emotional state (personality inventories). Verbal and numerical comprehension was measured post-operatively following adaptation of the speech processor. Performance and comprehension data were correlated. RESULTS: Cognitive performance parameters did not differ significantly from those of a healthy group. There was a correlation of r = +0.65 between numerical comprehension and the "mosaic-test", a subtest of the WAIS and of r = +0.78 between numerical comprehension and the crystallized intelligence scale. CONCLUSIONS: Pre-operative cognitive parameters may predict the outcome of cochlear implants. PMID- 11005090 TI - [Differential diagnosis of sinugenic enophthalmos]. AB - BACKGROUND: Spontaneous enophthalmos without recent trauma is a rare condition. Its origin is difficult to evaluate. METHOD: Specific properties of this symptom complex are presented based on a literature review and on case reports. Possible connections between enophthalmos and paranasal sinus diseases as well as differential diagnoses are analysed. PATIENTS: 1. 31 year old female patient with right spontaneous enophthalmos and no history of trauma. CT-imaging disclosed tissue formation in the maxillary sinus, partially destroyed medial orbital wall and floor, descended orbital contents following Caldwell-Luc procedure several years previously. After endonasal surgery of ethmoidal and maxillary sinus with removal of a large cyst good functional and cosmetical result. 2. 25 year old male patient complaining of pain in the periorbital region, presenting with left enophthalmos and superonasal deviation of the eyeball. MRI and CT revealed a tumor in the orbital floor region with total destruction of the roof of the maxillary sinus. Removal of the tumor by a combined lateral rhinotomy and subciliary approach. DIAGNOSIS: Leiomyoma. Orbital reconstruction with PDS-sheet. Postoperatively, improved globe position and motility. No recurrence during three year follow-up. CONCLUSION: When evaluating the causes of enophthalmos, chronic diseases of the paranasal sinuses or their walls must be considered. Surgical therapy is promising. PMID- 11005091 TI - [Long-term outcome of frontal sinus surgery: comparison of extranasal and endonasal surgical techniques]. AB - BACKGROUND: During the last decades rhinosurgery largely developed up to endonasal accesses. Meanwhile many of the new techniques are established, so that the outcome of traditional extranasal methods should be compared with those of endonasal accesses, in order to estimate the status and for critical consideration. METHODS: A retrospective analysis recorded the results of 236 patients, operated on primary frontal sinus disease 1985-1993. The records were consulted concerning inpatient treatment and outpatient aftercare during standardized paranasal sinus consulting hours. RESULTS: During the follow-up period (3-10 years) 8% of the patients underwent resurgery. The lowest revision rate was found after using endonasal technique (5.9%), compared with osteoclastic methods (Ritter-Jansen, Riedel: 10.6%). The domain of osteoplastic operations with a revision rate of 9.4% were fractures, osteomas and mucoceles, because of an impossible endonasal access. In decision on osteoclastic procedures, the already osteoclastically operated site was of prime importance, especially since 1990. CONCLUSION: Already during the early years endonasal techniques of paranasal sinus surgery showed better outcome than osteoclastic accesses. The analysis indicates that the change to endonasal functional paranasal sinus surgery is justified and should be kept up. The traditional methods still have their rare authorization in case of special indications, especially if an osteoclastic procedure has been carried out already. PMID- 11005092 TI - [hBD-2 gene expression in nasal mucosa]. AB - BACKGROUND: Chronic sinusitis is one of the frequent inflammatory diseases and has a complex pathogenesis. A substantial factor seems to be recurrent bacterial infections. Pseudomonas aeruginosa (PA) frequently can be found in nasal smears of patients with persistent sinus symptoms after sinus surgery. Lately a new antimicrobial peptide of epithelial origin, human Beta-Defensin-2 (hBD-2), with a strong antibacterial effect against PA could be identified within lesional skin scales of patients suffering psoriasis. Aim of this study was to investigate hBD 2-mRNA expression in nasal cells and tissue. METHODS: Total RNA was extracted from nasal polyps and turbinates following TRIzol protocol. Epithelial cells and fibroblasts of nasal human tissue were isolated and cultivated. The cells were stimulated with PA using different time points and different concentrations. Total RNA was isolated as mentioned above, reverse transcribed and amplified in a Semi-quantitative Reverse Transcriptase PCR (SQRT-PCR) with genespecific hBD-2 primers. RESULTS: PA induces time- and dose-dependently hBD-2 gene expression in nasal epithelial cells. Unstimulated epithelial nasal cells were able to express hBD-2 mRNA constitutively, whereas nasal fibroblasts showed no hBD-2 mRNA expression. Nasal polyps showed a comparable less hBD-2 gene expression then nasal turbinates. CONCLUSIONS: hBD-2 possibly mediates a specific, early starting antimicrobial defense strategy of the nasal mucosa. This hypothesis would explain persistent infections with PA through diminished hBD-2 gene expression. PMID- 11005093 TI - [Neuronavigation in the region of the skull base]. AB - Neuronavigation (computer-aided surgery planning and performance) has proven to be helpful in performing neurosurgical operations. The experiences of our department gained on more than 500 patients operated upon will be presented, with special focus on more than 100 operations for lesions on the skull base. Navigation is one of several modern surgical tools, its value cannot be defined without observing other improvements e.g. in the area of preoperative diagnostics, intraoperative monitoring, microsurgical instruments and microsurgical approaches. PMID- 11005094 TI - [The joint capsule of the cricoarytenoid joint: biomechanical and clinical aspects]. AB - BACKGROUND: Impaired movement of the cricoarytenoid joint with hoarseness and immobility of the vocal ligament may occur as a consequence of laryngeal trauma and joint disease. Little is known to date about the cricoarytenoid joint capsule and its role in joint pathology. METHODS: The present study analyses the structure of the cricoarytenoid joint capsule by means of histological, immunohistochemical, and scanning electron microscopical methods. Investigations are performed on larynges of 17 male and 16 female. RESULTS: The cricoarytenoid joint was found to be lined by a wide and lax joint capsule consisting of a fibrous and a synovial membrane. The capsule was strengthened posteriorly by the cricoarytenoid ligament. As like the fibrous membrane the cricoarytenoid ligament consisted mainly of collagen types I and III. Moreover the ligament was found to be rich in elastic fibers. Unexpected large and intensively vascularized synovial folds projected into the joint cavity. CONCLUSION: The capsule of the cricoarytenoid joint can be compared with the joint capsules of the limbs despite its structure and its involvement in joint pathology. Based on the laxity of the joint capsule it was concluded that invasive interventions at the respiratory tract with dislocation of the arytenoid cartilage can lead to incarceration of the synovial folds. After a trauma aero-synovitis or formation of hemarthrosis may occur, with subsequent fixation of the arytenoid in an abnormal position. PMID- 11005095 TI - [Transoral treatment of severe laryngomalacia. Review and presentation of a modified surgical technique]. AB - BACKGROUND: Laryngomalacia is the most common cause of stridor in children. So far various authors have described surgical techniques in a sense of a supraglottoplasty or an epiglottoplasty to handle the severe form of this disease. We present a modified technique of the so called epiglottopexy. METHODS: Under the different types of laryngomalacia this study focuses on the treatment of three patients (5 to 10 months) with a posterior displacement of the epiglottis during inspiration. The first step of our technique was to denude a small portion of the base of the tongue from mucosa. A corresponding lesion was created on the lingual surface of the epiglottis. The vaporisation of the mucosa was performed with the CO2 laser at a power setting of 1 Watt and a 0.25 mm spot size. The epiglottis was then fixed transorally to the base of the tongue with 2 single stitch sutures and 1 inverse mattress suture, using a resorbable material. RESULTS: No intra- or postoperative complications were observed. All three patients demonstrated a significant airway improvement without stridor. CONCLUSION: The presented technique of transoral laser surgical epiglottopexy seems to be a method suitable for the treatment of laryngomalacia caused by posterior displacement of the epiglottis. PMID- 11005096 TI - [Precancer stages of the oral mucosa: a review]. AB - According to the WHO collaborating centre precancerous lesions and precancerous conditions have to be distinguished. Precancer: BACKGROUND: Erythroplakia is the most dangerous precancerous lesion. It is rare, but may often remain undetected. It will transform into cancer within five years and therefore, has to be excised in every case. Leukoplakias show malignant transformation in 3-45% of the cases. In spite of modern molecular biological and immunohistochemical techniques the clinical appearance and the histological grading of the dysplasia are still most important prognostic factors. Until 1992 every lesion showing signs of moderate and severe dysplasia was excised in our department. Despite this treatment strategy 6.2% of the leukoplakias (n = 161) transformed into cancer. Therefore, we recommend to remove every lesion which does not disappear after eliminating the etiological factors. METHODS: Since 1992 168 leukoplakias were completely removed using the CO2 Laser and underwent histological examination. RESULTS: In 3% of these cases a carcinoma was detected in the leukoplakia; 5% of the lesions recurred. Precancerous condition: The most important precancerous condition, the oral lichen planus is treated in cases of erosive lesions only or if the patient is suffering from the symptoms. Malignant transformation is seen in 1.5% of the patients within 10 to 15 years. Histologically the oral lichen planus does not differ from the oral lichenoid reactions, lesions in contact with amalgam restorations mostly. In these cases a causative treatment with replacement of the amalgam is recommended. PMID- 11005097 TI - [Surgical and adjuvant drug therapy in head and neck cutaneous melanoma]. AB - The rapid incidence rise of cutaneous melanoma resulted in an increasing interest in this particular tumor. During the last years public prevention campaigns enlarged the awareness of melanoma, subsequently as a direct effect the mean tumor thickness of melanoma, the most predictable prognostic factor, decreased. Moreover, the biology of melanoma initiation and metastasis has been studied extensively with special interest in molecular biology. Controlled clinical studies answered several critical questions in respect to the standard care of surgery in melanoma. Yet, the guidelines for the surgical treatment of head and neck melanoma are in accordance to that of other localisations with reduced safety margins around the primary tumor. Elective (prophylactic) lymph node dissection (ELND) of regional lymph nodes is no more considered as a standard tool. Moreover, ELND has been given up by most melanoma centers, since it is known that prospective-randomized trials were not able to demonstrate an increase of overall survival for patients with ELND compared with untreated patients. Instead of this potentially aggressive treatment modality the examination of the first draining regional lymph node, sentinel node biopsy (SNB), has been introduced some years ago. Recently, a large clinical trial demonstrated that the SNB status reflects the most valuable prognostic factor for primary melanoma known so far. First studies in head and neck melanoma figured out that this technique is more complex in this special localisation, but produced comparable results. Systemic adjuvant (prophylactic) therapy of high-risk melanoma should preferentially be applied within controlled clinical trials. Most attractive candidates for an effective treatment are interferons. Several studies ruled out that interferon alpha-treated melanoma patients demonstrate an extended disease free survival. Adjuvant chemotherapy has not shown a clinically relevant benefit. Thus, patients should preferentially be treated within controlled clinical trials. PMID- 11005098 TI - [Analysis of the p53 gene status of lymph node metastasis in the head and neck region in occult primary cancer]. AB - BACKGROUND: Lymph node metastasis in the head and neck region with occult primary cancer are a fairly rare tumor entity in head and neck cancer. Tumor biological parameters as well as mechanism and cause for the development of this so called "cancer of unknown primary" (CUP) are not well investigated in head and neck cancer. Mutations of the p53 tumor suppressor gene are the most prevalent genetic alteration of human malignancies and show an incidence of up to 50% in head and neck cancer. For a further tumor biological definition of CUP the p53 status was determined. METHODS: Twenty-three archival formalin fixed paraffined CUP of an occult squamous cell carcinoma of the head and neck region were examined. In all cases a primary cancer was never diagnosed. The DNA extracted from the tumor material was amplified with specific primers for exon 4-9 of the p53 gene and subsequently sequenced. RESULTS: None of the 23 CUP cases showed a mutation or polymorphism in exon 4-9 of the p53 gene. CONCLUSIONS: The total absence of p53 mutations in the so called mutational "hot spots" shows a significant difference to the frequency of primary tumors of the head and neck region. May be this tumor biological characterisation helps to further elucidate the growth behaviour of CUP in the head and neck region and the reason for their development. PMID- 11005099 TI - [Detection of overexpression of insulin receptor gene in laryngeal carcinoma cells by using differential display method]. AB - BACKGROUND: The insulin receptor (IR) is an essential protein localized on the surface of almost all cell types. IR belongs to the tyrosine-kinase growth factor receptor family. Insulin mediates proliferative responses in a variety of tumor cells, however, the role of the IR molecule in carcinogenesis has not yet exactly been established. METHODS: Messenger RNA from mucosal keratinocytes and laryngeal carcinoma cells were transcribed in cDNA using reverse transcriptase and amplified by PCR by means of a number of oligonucleotides. PCR products were analyzed electrophoretically. RESULTS: Comparing the electrophoretic pattern of both cell types the overexpression of a 127 bp fragment could be detected in laryngeal carcinoma cells in contrast to benign keratinocytes. Cloning and sequencing this fragment exact homologous match was found with exon-2 of the IR gene. The overexpression of the IR gene in laryngeal carcinoma cells was confirmed by Northern hybridization. CONCLUSIONS: The results show up-regulation of IR-mRNA in laryngeal carcinoma cells suggesting that the number of IR is enhanced in these cells. Hence it follows, that the overexpression of IR plays a possible role in laryngeal cancer initiation and/or progression. PMID- 11005100 TI - [Feeding of patients with ENT diseases using an enteral tube system (PEG/PEJ tubes). A review]. AB - In patients with benign or malignant diseases of the larynx/pharynx causing inadequate oral food intake, maintenance of individual adequate enteral nutrition is a major medical goal. This is particularly important in patients with malignant ENT tumours, where therapeutical measures as radiation or chemotherapy result in further deterioration of nutritional status and consecutively of quality of life and tolerance of therapy. Due to its safe and technically simple route of placement, the PEG has become the preferred enteral route of choice for enteral long-term nutrition world-wide. The technical success rate is > 99% with a procedure-related lethality rate of about 0%. Prospective clinical studies prove the excellent individual subjective acceptance of this method by patients. Indications, contraindications, complications, technical procedures and clinical aspects of PEG placement are described in detail in the present review. Long-term enteral feeding via PEG is accepted as a safe, effective, easy to practise and highly acceptable method with excellent long-term results and thus distinct improvement of nutritional status. Individual decision for PEG placement should be considered much earlier and more frequently in appropriate patients to save and further improve clinical and nutritional status and consecutively quality of life. PMID- 11005101 TI - [The interesting case No. 36. Fibrous dysplasia of the temporal bone (osteodystrophia fibrosa Jaffe-Lichtenstein)]. PMID- 11005102 TI - [Plastic reconstructions in the neck region. Neck surface soft tissue and skin defects. I]. PMID- 11005103 TI - Use of an emboli containment and retrieval system during percutaneous coronary angioplasty in native coronary arteries. AB - BACKGROUND: Prevention of distal embolisation during percutaneous coronary revascularisation may be necessary to reduce postinterventional morbidity and mortality. METHODS AND RESULTS: We employed a newly developed emboli containment and retrieval system in native coronary arteries during percutaneous coronary angioplasty and stenting in 39 selected patients (mean age 58.9 +/- 10.1 years, 11 females) presenting with acute (n = 22; 8 LAD, 3 LCX, 11 RCA), subacute (n = 7; 2 LAD, 2 LCX, 3 RCA) or chronic (n = 6; 2 LAD, 4 RCA) total or subtotal occlusion of an infarct-related vessel, or with severe stenosis and symptoms of unstable angina (n = 4; 2 LAD, 2 RCA). Protection device-assisted angioplasty with stent implantation was uneventful in all patients with good angiographic results and normal postprocedural flow. Intermittent aggravation of anginal pain during inflation of the occlusive balloon (from 2.5 to a maximum of 25 minutes cumulative inflation time) was observed in 19 of the 36 conscious patients (7 with acute, 7 with subacute and 3 with chronic occlusion, and 2 with unstable angina), but caused neither interruption of distal occlusion nor haemodynamic instability. In 31 patients the aspirates contained visible debris. Histological analysis showed particles up to 12 mm in size, consisting of necrotic core, inflammatory cells, cholesterol debris, and old and fresh thrombi. In 8 patients the aspirated particles were too small to allow microscopic diagnosis or debris was absent. CONCLUSIONS: This preliminary report demonstrates the feasibility of using a protection device in native coronary arteries to prevent distal embolisation of particulate matter that is mobilised during percutaneous interventions. To the extent that this material contributes to the mechanisms of distal embolisation, noreflow and infarction, this device may help to reduce such complications. Appropriately designed trials are required to assess the clinical benefit of this system. PMID- 11005104 TI - [Cost-effectiveness of primary PTCA and thrombolysis in the treatment of acute myocardial infarction]. AB - Clinical studies have shown a favourable outcome for primary PTCA compared with thrombolysis in the treatment of acute myocardial infarction. No data are available in Switzerland on the logistic and economic implications of treating more acute myocardial infarction patients by PTCA. The present paper sets out to assess all published studies comparing the cost-effectiveness of the two treatment modalities. A Medline search identified seven original cost and cost effectiveness studies conducted between 1989 and 1999. According to these studies emergency PTCA generates costs similar to thrombolysis in the treatment of acute myocardial infarction if the infrastructure is available and there is high volume output. Better clinical results, as suggested by the literature, would result in a favourable cost-effectiveness ratio for primary PTCA. PMID- 11005105 TI - Manifold manifestations of ergotism. AB - Vasospastic side effects leading to organic manifestations are rare in ergotamine therapy. To our knowledge, combinations of more than two signs of ergotism have rarely been described in the literature so far. We present a 65-year-old male patient who as a consequence of severe migraine had developed ergotamine abuse. He was admitted to our hospital after one week of increasing abdominal pain. During laparotomy, necrotic areas of the small intestine and the sigmoid colon were resected, which on histopathologic examination revealed severe hypertrophy of the smooth musculature of mesenteric arteries, resulting from chronic vasospasms. Postoperatively, the patient developed ischaemia of the limbs which was confirmed by angiography. Before death, the patient also showed ischaemic signs in the acrae and necrosis of the tongue. PMID- 11005106 TI - [Cholesterol and stroke risk: a role for statins?]. AB - Atherosclerosis is the most common cause of vascular diseases, but the relevance of cholesterol has only been definitely associated with coronary artery disease and peripheral vascular disease. In comparison, the role of cholesterol in stroke is, while a tempting assumption, subject to controversy in the literature. The crucial question--is cholesterol a risk factor for stroke?--remains open. Recent trials with statin drugs, such as 4 S, CARE, LIPID and WOSCOP, have created a new wave of enthusiasm by showing decreased risk of stroke in the statin-treated patients. However, these trials are most often designed for patients with a known history of coronary artery disease. In contrast, studies investigating the impact of statins in the secondary prevention of stroke are still lacking. Moreover, the beneficial effects of statins on clinical events may involve non-cholesterol mechanisms. In regard to stroke prevention, there is no absolute evidence to recommend the use of statin drug therapy. PMID- 11005107 TI - [Tropical air in Zurich]. PMID- 11005108 TI - [Reference: Franke C. Value of radiosynovectomy in rheumatology and orthopedics]. PMID- 11005109 TI - Should sonographic screening for fetal Down syndrome be applied to low risk women? PMID- 11005110 TI - Assessment of hemodynamics using Doppler ultrasound. PMID- 11005111 TI - The frequency of the detection of fetal echogenic intracardiac foci with respect to maternal race. AB - OBJECTIVE: To determine if there is a racial difference in the frequency of identification of echogenic intracardiac foci (EIF) seen sonographically in the hearts of second-trimester fetuses. METHODS: Over a 2-month period (June 1998 August 1998), all fetuses scanned between 15 and 20 completed weeks' gestation were evaluated prospectively for the presence or absence of EIF. Pregnancies specifically referred for the presence of EIF were excluded. The sonographer performing the scan indicated maternal race as Asian, black, white, or (if maternal race was not clear) unknown. Maternal race, gestational age, and the presence or absence of EIF were prospectively documented. Follow-up of those fetuses with EIF was obtained from the referring physicians' offices. The groups were compared with respect to maternal race and presence or absence of EIF. RESULTS: There were 46, 34, 400, and nine fetuses of the Asian, black, white, and unknown mothers, respectively. The mean gestational age +/- 1 SD at examination was 18.2 +/- 1.6, 17.5 +/- 1.4, 17.7 +/- 1.5, and 17.8 +/- 1.1 weeks, for the Asian, black, white, and unknown mothers, respectively. The incidence of sonographically detected EIF was 30.4, 5.9, 10.5 and 11.1% for the Asian, black, white, and unknown mothers, respectively, P = 0.001. In a multivariate logistic regression model, Asian mothers had an odds ratio of 3.8 (95% CI, 1.8, 7.6) for having a fetus identified as having EIF, as compared with white mothers. CONCLUSIONS: The Asian patient is more likely than patients of other races to have a fetus with identified EIF. The counseling implications for Asian mothers undergoing midtrimester sonography when EIF is identified should be tempered, due to the increased frequency of EIF as a normal finding in the Asian population. PMID- 11005112 TI - Color Doppler imaging in the diagnosis and management of chorioangiomas. AB - Color Doppler imaging was used to investigate nine consecutive cases of placental tumors referred for evaluation to a tertiary referral unit. Gray-scale ultrasound findings were of a chorioangioma which was subsequently confirmed on pathologic examination. On color Doppler imaging (CDI), three tumors appeared avascular and the pregnancies were uncomplicated, two had only a few vessels in their periphery, and four contained numerous vessels. These vascular chorioangiomas were complicated by polyhydramnios and premature labor (n = 3), isolated polyhydramnios (n = 1), fetal growth restriction (n = 1) and generalized non immune fetal hydrops (NIHF) (n = 1). The maximum diameter of the tumors ranged between 3 and 10 cm. There was no direct association between the size and location of the tumor and the development of complications. Successful amnioreduction was performed in the three cases of polyhydramnios and premature onset of labor. Vascular sclerosis was attempted unsuccessfully in the case complicated by severe NIHF. Our data indicate that the vascularization of the tumor is a pivotal determinant factor of pregnancy outcome. Where the tumor is avascular, no specific complications should be expected. Where the tumor is vascularized, and in particular if it contains numerous large vessels, serial ultrasound and Doppler examinations are warranted to detect polyhydramnios and early features of fetal congestive heart failure. PMID- 11005113 TI - The coronary sinus in the fetus. AB - INTRODUCTION: The normal coronary sinus drains venous blood from the cardiac veins to the right atrium. In some instances, the coronary sinus may be dilated due to volume or more rarely pressure overload. AIMS: To assess the feasibility of detecting the coronary sinus in the fetus and to establish the normal values of the coronary sinus dimensions throughout gestation. SUBJECTS AND METHODS: Fetal echocardiography was performed in 78 normal fetuses from the 16th to the 40th week of gestation (median 25 week). The coronary sinus was measured in four chamber view (87% of cases) or in parasternal short axis view equivalent (13%). A second group of nine fetuses with a dilated coronary sinus was compared to the normal group. RESULTS: Adequate imaging of coronary sinus was obtained in 97.4% of the normal fetuses. The diameter of the coronary sinus ranged from 1 to 3.2 mm (2 mm +/- 0.13 mm, mean +/- 5% confidence interval) and correlated well with the age of pregnancy (r = 0.86). The length-to-diameter ratio of 24% (+/- 6%) did not vary throughout pregnancy. All nine fetuses with a dilated coronary sinus had a persistent left superior vena cava which drained into it. The diameter of the coronary sinus was approximately three times larger in the abnormal group with a diameter-to-length ratio of approximately 83% (P < 0.0001). CONCLUSION: The coronary sinus is readily identified in the fetus. It gradually increases during pregnancy. An abnormal coronary sinus is easily diagnosed and should prompt the sonographer to look for a persistent left superior vena cava. PMID- 11005114 TI - Routine ultrasound fetal examination in pregnancy: the 'Alesund' randomized controlled trial. AB - OBJECTIVE: To evaluate the possible benefits of the routine use of ultrasound screening in pregnancy. DESIGN: A randomized controlled trial was designed to detect a 50% difference in the incidence of induction for apparent post-term pregnancies between women who were screened with ultrasound and unscreened women. A total of 1628 pregnant women from the general population were included. Eight hundred and twenty-five were allocated to an ultrasound examination at the 18th and 32nd week of pregnancy in addition to receiving routine antenatal care. The remaining 803 women received standard antenatal care, but could only be referred for ultrasound examination on clinical indication. RESULTS: The incidence of induced labor due to apparent post-term pregnancies was approximately 70% lower in the ultrasound-screened group. Inductions from all causes were also less frequent among ultrasound-screened women. There were six perinatal deaths among the screened and seven among the controls after excluding three lethal malformations among the controls. There was no difference in Apgar score after 1 min, but the proportion with Apgar score less than 8 after 5 min was lower among the screened group (P = 0.04). The need for positive pressure ventilation for more than 1 min was lower among the screened group (P = 0.02). Birth weight was slightly higher in the screened group (39 g), but the difference was not statistically significant. Among the controls three pairs of twins remained undiagnosed until the mothers were admitted to the hospital in labor at between 36 and 38 weeks gestation. CONCLUSION: These results suggest that for women who were screened with ultrasound, obstetricians were less likely to induce labor due to apparent post-term pregnancy, than for women who were not screened. All 10 pairs of twins in the screened group were diagnosed at the routine examination. These data also suggest that perinatal morbidity might be slightly lower in the screened group. PMID- 11005115 TI - Human fetal pulmonary artery velocimetry: repeatability and normal values with emphasis on middle and distal pulmonary vessels. AB - OBJECTIVES: To establish the nature and gestational age dependency of flow velocity waveforms from fetal middle and distal arterial pulmonary branches in the second half of normal pregnancy and to determine repeatability and inter relationship of flow velocity waveform recordings from proximal, middle and distal arterial pulmonary branches. DESIGN: Cross-sectional study. SUBJECTS/METHODS: A total of 111 singleton normal pregnancies between 20 and 40 weeks of gestation were studied using a color-coded Doppler ultrasound system. Pulmonary waveforms were obtained at the level of the fetal cardiac four-chamber view. Repeatability was tested from two recordings at 15 min time-intervals in 25 separate normal pregnancies. RESULTS: Acceptable repeatability of flow velocity waveforms from fetal arterial pulmonary branches was established with coefficients of variation below 15%. The nature of middle arterial pulmonary flow velocity waveforms was similar to that of proximal waveforms and showed a gestational age-related change for diastolic velocity parameters, peak systolic/peak diastolic ratio and pulsatility index. The distal arterial pulmonary branch displayed a monophasic forward flow velocity profile throughout the cardiac cycle. All velocity parameters of the distal branch remained unchanged with advancing gestation, with the exception of the pulsatility index. Significant inter-pulmonary changes were found for all pulmonary arterial waveform parameters. CONCLUSIONS: Alteration in pulmonary vascular resistance may play a role in gestational age-related changes, whereas changes in vessel branching/diameter and in the distance between the heart and more distal arterial pulmonary vessels may cause inter-pulmonary differences. PMID- 11005116 TI - Maternal cardiac systolic and diastolic function: relationship with uteroplacental resistances. A Doppler and echocardiographic longitudinal study. AB - OBJECTIVE: To test the hypothesis of the existence of a relationship between central and peripheral hemodynamic parameters by the longitudinal evaluation of maternal echocardiographic and uteroplacental resistance modifications during normal pregnancy. METHODS: Forty-three healthy normotensive primigravidae were evaluated at 12 +/- 1, 21 +/- 1, and 33 +/- 1 weeks of gestation with uterine artery color Doppler and maternal echocardiographic examinations to identify morphologic, systolic, and diastolic variables. RESULTS: Cardiac output and stroke volume significantly increased during pregnancy. Uterine resistance index (RI) decreased from the first to the second trimesters (0.72 +/- 0.10 versus 0.54 +/- 0.09, P < 0.001). Left atrial dimensions increased during pregnancy (33.8 +/- 1.9 cm, 38.1 +/- 1.8 cm, 39.3 +/- 2.1 cm, P < 0.001). Left atrial function also increased. Left ventricular mass increased (132 +/- 18 g, 162 +/- 16 g, 174 +/- 27 g, P < 0.001). Diastolic function parameters showed significant modifications: E wave velocity and E/A ratio decreased; A wave velocity and deceleration time of the E wave (DtE) increased; the left ventricular isovolumetric relaxation time (IVRT) decreased significantly (88.7 +/- 6.7 ms, 75.6 +/- 7.7 ms, 71.1 +/- 5.0 ms, P < 0.001) showing a correlation with left atrial dimensions and RI (r = 0.38, r = 0.47, respectively; P < 0.001). CONCLUSIONS: Diastolic cardiac function varies during pregnancy. A relationship between preload (left atrial enlargement), afterload (RI reduction), morphologic, and diastolic function modifications (IVRT reduction, DtE prolongation) appears to exist as a consequence of the hemodynamic modifications which occur during physiologic pregnancy. Diastolic function analysis maybe useful to identify women who fully adapt to pregnancy, and to understand the mechanisms that might be involved in women who show abnormal uterine artery Doppler waveforms. PMID- 11005117 TI - The influence of maternal exercise on placental blood flow measured by Simultaneous Multigate Spectral Doppler Imaging (SM-SDI) AB - OBJECTIVE: To evaluate the effect of maternal isometric exercise on the placental blood flow as reflected by the velocimetric indices PI and RI derived from placental arteries. SUBJECTS: Thirty-four healthy women with normal singleton pregnancies between 22 and 35 weeks of gestation. METHODS: All subjects underwent an isometric handgrip exercise test. Maternal blood pressure and heart rate together with placental PI and RI were measured at rest, during the exercise and in the post-exercise recovery phase. All Doppler measurements were obtained using the Simultaneous Multigate Spectral Doppler Imaging (SM-SDI) technique, a new ultrasound modality that enables a Doppler study of multiple locations to take place within a very short time. RESULTS: There was a significant increase in the mean values of the maternal blood pressure and heart rate during the exercise and a significant decline in the recovery phase. There was no significant change in the mean values of the Doppler indices throughout the examination. CONCLUSION: Isometric handgrip exercise test during pregnancy does not affect the impedance of the placental circulation. PMID- 11005118 TI - Non-invasive assessment of endothelial function in normal pregnancy. AB - OBJECTIVE: To assess endothelial function in normal pregnancy by non-invasive methods. METHODS: Flow-mediated dilatation of the brachial artery was measured by ultrasonography in 157 women with normal singleton pregnancies between 10 and 40 weeks' gestation and 19 non-pregnant controls. RESULTS: Flow-mediated dilatation in the non-pregnant controls was 6.42 +/- 2.45%. In pregnant women, between 10 and 30 weeks, the mean flow-mediated dilatation (8.84 +/- 3.18%) was significantly higher than the non-pregnant controls (P = 0.002), but after 30 weeks of gestation there was a decrease to prepregnancy levels. Resting vessel diameter and blood flow were significantly increased in pregnancy, mainly after 30 weeks' gestation (P < 0.001, P < 0.001, respectively). Flow-mediated dilatation was significantly correlated to resting vessel diameter and reactive hyperemia. CONCLUSION: Normal pregnancy is associated with enhanced endothelial function which is apparent from at least 10 weeks' gestation. PMID- 11005119 TI - Automated cardiac output measurements by ultrasound are inaccurate at high cardiac outputs. AB - OBJECTIVE: The sonographic technique of automated cardiac output measurement (ACM) is a promising new method to measure cardiac output and could be of use in a high-risk obstetric unit in the treatment of pre-eclamptic patients. The aim was to determine the accuracy of the ACM method. DESIGN: Comparative study of the sonographic technique of ACM versus cardiac output measured by thermodilution (TD). METHODS: The study included 39 intensive care patients, 21 men, 13 non pregnant women and five severely pre-eclamptic pregnant patients, with a wide range of cardiac outputs, in whom TD catheters had been inserted for clinical reasons. Two separate experienced observers, blinded to the results obtained with the other method, performed four successive measurements in each patient with either the ACM or TD technique. The averaged cardiac output value per patient and method was used for comparison. RESULTS: Cardiac output was successfully measured with ACM and TD in 85 and 100% of patients, respectively. Mean cardiac output measured by ACM (6.77 +/- 1.90 L/min) was significantly lower than that measured by TD (9.12 +/- 3.06 L/min). Although cardiac output values obtained with ACM were significantly correlated with those measured by TD, the ACM values were consistently lower than TD values in the higher cardiac output range; the relationship was represented by ACM = 0.35 TD + 3.55 L/min (r = 0.57, P < 0.001). The (ACM - TD) difference increased significantly with cardiac output, through a difference in stroke volume, not in heart rate. CONCLUSION: The ACM is not an accurate tool to measure cardiac output in patients with a high cardiac output, including treated pre-eclamptic women. PMID- 11005120 TI - The effect of methyldopa on retinal artery circulation in pre-eclamptic gravidae. AB - OBJECTIVE: To evaluate the effect of methyldopa on retinal artery circulation in pre-eclamptic gravidae using color Doppler imaging and spectral analysis. METHODS: Fifty-three pre-eclamptic singleton gravidae of gestational age greater than 22 weeks were examined. Patients with sustained hypertension after 1-2 days hospital rest were treated with oral antihypertensive medication, 250-500 mg methyldopa, three to four times a day for a minimum of 5-7 days. The right central retinal arteries were insonated and Doppler waveform values were analysed before and after medication. RESULTS: The change of the maternal heart rate after methyldopa treatment was -3.96 +/- 7.88 beats per min (P = 0.0006). The change of fetal heart rate was not significantly altered. The change of the diastolic arterial blood pressure after treatment was -4.19 +/- 12.36 mmHg (P = 0.0169). In 36 gravidae, in whom hypotensive effects were noted after treatment with methyldopa, the increase in peak velocity, end-diastolic velocity and mean velocity of the retinal artery were 2.41 +/- 2.20 (P < 0.0001); 1.48 +/- 1.23 (P < 0.0001) and 1.70 +/- 1.42 (P < 0.0001), respectively. The decrease in pulsatility index of the retinal artery after treatment with methyldopa was -0.17 +/- 0.22 (P < 0.0001). In the remaining 17 gravidae, in whom no hypotensive effects were noted after treatment with methyldopa, the decrease in end-diastolic velocity and mean velocity were -1.50 +/- 1.70 (P = 0.0022) and -0.98 +/- 1.90 (P = 0.0488), respectively. The increase in pulsatility index was 0.34 +/- 0.30 (P = 0.0003). CONCLUSIONS: In pre-eclamptic gravidae in whom the hypotensive effects were noted after treatment with methyldopa, the mean velocity of the retinal arteries was significantly higher and the mean pulsatility index lower after treatment. We conclude that the hypotensive effect of methyldopa in pre-eclamptic gravidae is associated with a significant decrease in retinal artery vascular resistance. PMID- 11005121 TI - The influence of fetal position on nuchal translucency thickness. AB - OBJECTIVE: To determine whether nuchal translucency thickness is influenced by the fetal position at ultrasound examination. SUBJECTS: Transabdominal ultrasound examination for pregnancy dating and measurement of nuchal translucency thickness was performed at 10-14 weeks' gestation in all women attending the antenatal clinic of our hospital. During the examination special attention was paid to a change in fetal position from prone to supine or vice versa. METHODS: For each fetus the nuchal translucency measurement was repeated when a positional change from prone to supine or vice versa was recorded. All measurements were recorded on hard copy. An image-scoring method was used and evaluated by three independent reviewers. RESULTS: Eighty-five fetuses were included in this study. The mean nuchal translucency for supine fetuses was 1.91 mm compared with 1.93 mm for prone fetuses. The mean quality-score was 6.54 for supine fetuses and 6.55 for prone fetuses. This difference was not statistically significant. CONCLUSION: Fetal position has no influence on the measurement of nuchal translucency. PMID- 11005122 TI - Sonographic visualization of normal-size ovaries during pregnancy. AB - OBJECTIVE: To assess the ability of ultrasound to detect ovaries of normal size during pregnancy METHODS: A prospective study of 329 women with a normal pregnancy course was undertaken; 68 were excluded from analysis because of an enlarged, cystic ovary. Of the remainder, 60 pregnancies were examined in the first trimester and 201 in the second or third trimester. The first group underwent transvaginal (TVS) and transabdominal (TAS) scanning. The second group underwent TAS examination only. RESULTS: In the first-trimester group, TVS identified both ovaries in 57 patients (95%) and transabdominal ultrasound in 20 (33.3%). In the second- and third-trimester patients, TAS visualized both ovaries in 33 patients (16.4%), and neither ovary in 120 (59.7%). Both ovaries were less visible with advancing gestational age. The right ovary showed a significant change in position during pregnancy, from about 1 cm (at 15-24 weeks) to 2.5 cm (at 30-41 weeks) cranial to the iliac spine. The left ovary was found 1 cm above the iliac spine throughout pregnancy. CONCLUSIONS: Transvaginal sonography is adequate for the visualization of both ovaries in the first trimester of pregnancy. With advanced gestational age, the ovaries were significantly less visible by TAS. Sonographic scanning of the ovaries in second and third trimester should be concentrated mainly at the level of the iliac spine. Poor sonographic visualization of both ovaries in late gestation may mandate the use of other imaging modalities. PMID- 11005124 TI - Increased fetal nuchal translucency: possible association with esophageal atresia. AB - We describe a case in which in-utero diagnosis of an esophageal atresia with a tracheo-esophageal fistula in the third trimester followed the finding of an increased nuchal translucency in the first trimester and suggest a mechanism by which these two findings might be associated. PMID- 11005123 TI - Uterine artery impedance to blood flow on the day of embryo transfer does not predict obstetric outcome. AB - OBJECTIVE: To evaluate the influence of uterine artery impedance to blood flow on the day of embryo transfer for prediction of early pregnancy loss and obstetric outcome. METHODS: The uterine artery pulsatility index (PI) and resistance index (RI) were evaluated prospectively by transvaginal Doppler in 102 infertile women, who conceived as the result of fresh or frozen embryo transfer. Uterine artery impedance to blood flow was compared to the obstetric outcome. RESULTS: The 111 treatment cycles studied resulted in 31 spontaneous abortions, four ectopic pregnancies, and 76 deliveries. There were no differences in uterine artery PI and RI (mean +/- SD) between cycles resulting in normal delivery (2.69 +/- 0.71 and 0.88 +/- 0.06) and those resulting in spontaneous abortion (2.71 +/- 0.67 and 0.88 +/- 0.05) or ectopic pregnancy (2.36 +/- 0.54 and 0.85 +/- 0.06). There were no differences in PI and RI between uncomplicated singleton pregnancies (2.74 +/- 0.78 and 0.88 +/- 0.06) and those developing intra-uterine growth restriction (IUGR), pregnancy-induced hypertension (PIH), or preterm birth (2.54 +/- 0.47 and 0.87 +/- 0.04, pooled data). CONCLUSIONS: Uterine artery PI and RI on the day of embryo transfer were unrelated to the risk of the pregnancy ending in spontaneous abortion or ectopic pregnancy. These values were of no value in the prediction of IUGR, PIH or preterm birth. PMID- 11005125 TI - Diagnosis of a fetal mesoblastic nephroma by 3D-ultrasound. AB - We report a case of a fetal renal tumor detected prenatally on 3D-ultrasound. As the lesion was well encapsulated the initial sonographic diagnosis was that of a nephroblastoma. Volume calculation by the 3D technique gave a reliable estimation of the tumor size. Contrary to all published case reports concerning antenatally diagnosed mesoblastic nephromas, there was no polyhydramnios. Elective delivery was performed by Cesarean section at 38+ weeks gestation. The neonate underwent left nephrectomy on the second day of life. The subsequent course was uneventful without recurrence of the tumor. PMID- 11005126 TI - Klippel-Trenaunay-Weber syndrome presenting as massive lymphangiohemangioma of the thigh: prenatal diagnosis. AB - We report a case of Klippel-Trenaunay-Weber syndrome presenting prenatally as a massive congenital lymphangiohemangioma of the thigh. Routine ultrasonographic examination revealed multiple distorted cystic areas extending from the right flank through the right lower extremity of a 30-week fetus. A diagnosis of cystic lymphangioma of the thigh was suspected prenatally. Neonatal evaluation confirmed the prenatal findings. Neonatal color Doppler imaging revealed blood vessels within the tumor. The differential diagnosis is discussed together with available therapeutic procedures. PMID- 11005127 TI - Acute bowel perforation in a fetus with gastroschisis. AB - Gastroschisis is a congenital anomaly with a reported incidence of 1 in 10,000 live births. Although prenatal diagnosis is more common with the widespread use of biochemical markers and obstetric ultrasound, the role of ultrasound in the identification of the fetus that might need early intervention has not been established. Acute bowel perforation was diagnosed by ultrasound at 34 weeks gestation in a fetus with gastroschisis. An immediate Cesarean section was performed, followed by repair with primary closure. The neonatal outcome was favorable. The post-partum findings, including bowel pathology, confirmed the antenatal diagnosis. Acute bowel perforation can be diagnosed antenatally. Immediate intervention, before further bowel injury occurs, might enhance the ability of the surgeon to perform primary closure and obtain a favorable outcome. PMID- 11005128 TI - Prenatal sonographic features of vesicoallantoic cyst. PMID- 11005129 TI - Ultrasound--frequently asked questions. PMID- 11005130 TI - Intrapericardial teratoma. PMID- 11005131 TI - [Human amniotic fluid transferrin stimulates progesterone production by human trophoblast cells in vitro]. AB - OBJECTIVE: During pregnancy transferrin plays a key role as an iron transport protein to serve the increased fetal demands of iron. Transferrin is also present in relatively high concentrations in amniotic fluid [6], showing a different glycosylation compared with serum transferrin. The biological function of human amniotic fluid transferrin (hAFT) is still unknown. In addition trophoblast cells also synthesise transferrin. Transferrin synthesised by the trophoblast shows a special glycosylation. We found identical carbohydrate structure of hAFT and trophoblast transferrin. We investigated the influence of hAFT on the progesterone-, cortisol- and hCG-release of trophoblasts in culture compared with the influence of human holo- and apo-serum transferrin on the release of these hormones. MATERIAL AND METHODS: Cytotrophoblast cells were prepared from human term placentae by standard trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation step. When placed in culture, the trophoblasts were incubated with varying concentrations (50-300 micrograms/ml) of human amniotic fluid- and serum-transferrin. Unstimulated cells of each placenta used as controls. Culture supernatants were assayed for progesterone, hCG and cortisol by enzyme-immunometric methods. RESULTS: Our results show, that the release of progesterone increased in hAFT-treated cell cultures compared to untreated cell cultures. Holo- and apo-serumtransferrin did not show any effect on the progesterone release by trophoblast cells in vitro. Neither hAFT nor holo- and apo-serum transferrin had any effect on the cortisol- and hCG-release in vitro. CONCLUSIONS: Progesterone is a marker for differentiation of trophoblasts in syncytiotrophoblasts. Only hAFT stimulates the progesterone production. We suggest, that hAFT can modulate the endocrine function of trophoblast cells in culture by regulating progesterone production. PMID- 11005132 TI - Determination of cytokine mRNA-expression in term human placenta of patients with gestational hypertension, intrauterine growth retardation and gestational diabetes mellitus using polymerase chain reaction. AB - OBJECTIVE: Our objective was to test the hypothesis, that pregnancy-related diseases are going along with changes in cytokine mRNA-expression at the placental site, either as a part of a pathological process or in connection with regulatory mechanisms induced by disturbances at the feto-maternal interface resulting from previous pathological changes--in the sense of counterregulation. MATERIAL AND METHODS: The cytokines chosen for this investigation are known to 1.) be expressed in the human placental tissue, 2.) to be involved in immunological processes and 3.) the regulation of growth and differentiation processes of different cell types of the placenta or decidua, 4.) to play a role in the angiogenesis at the feto-placental interface and 5.) to be involved in pathological processes in other human diseases. 32 samples derived from term human placentas were examined for messenger RNA levels of interleukin 1 alpha (II 1 alpha), tumor necrosis factor-alpha (TNF-alpha), platelet derived growth factor A chain (PDGF-A), platelet derived growth factor-B chain (PDGF-B), and platelet derived growth factor receptor (PDGF-R) using a semiquantitative reverse transcriptase (RT) polymerase chain reaction (PCR) protocol. To calibrate samples in our procedure, beta-actin mRNA (messenger ribonucleic acid) known as a "house keeping" gene was proven to be constantly expressed. The sample-groups consisted of normal pregnancies (n = 8), gestational hypertension (GH, n = 7), intrauterine growth retardation (IUGR, n = 6), gestational diabetes mellitus (GDM, n = 5), and gemini (n = 3 x 2). RESULTS: Throughout the 32 samples, a significant correlation between PDGF-A and PDGF-R expression, PDGF-A and TNF-alpha expression was stated (p = 0.007). Compared with the pattern of expression in normal placentas, placentas of growth retarded pregnancies had higher Il-1 alpha mRNA (p = 0.016), PDGF-A (p = 0.029) and PDGF-B (p = 0.001) levels. The samples of the gestational hypertension group and placentas of patients with gestational diabetes displayed a significantly stronger PDGF-R mRNA signal (p = 0.0029 and p = 0.008). CONCLUSIONS: Though these marked differences in cytokine mRNA levels between clinical groups were statistically proven, clear correlation of these differences with clinical data was not found. PMID- 11005133 TI - [Secondary malignant neoplasms in patients with breast cancer]. AB - OBJECTIVE: This report analyzes the occurrence of secondary malignancies among patients with breast cancer. MATERIAL AND METHODS: We evaluated all women diagnosed with breast cancer who were reported to the National Cancer Institute of the former GDR in 1976. There was a follow up of 5,485 patients from 1976 to 1988 (38,231 person-years at risk). Cancer incidence rates of the whole female population specific for age and calendar year were used for calculation of the standardized incidence ratio (SIR). RESULTS: Significantly increased risks were observed for neoplasms of the bilateral breast (SIR 2.44; 95% CI 2.04-2.93), colon (SIR 1.53; 95% CI 1.06-2.12), rectum (SIR 1.65; 95% CI 1.10-2.40), endometrium (SIR 1.55; 95% CI 1.02-2.27), and ovary (SIR 1.71; 95% CI 1.09-2.57). Neoplasms of the lung (SIR 1.65; 95% CI 0.93-2.73), kidney (SIR 1.51; 95% CI 0.69 2.87), bladder (SIR 1.32; 95% CI 0.48-2.87), connective tissue (SIR 3.54; 95% CI 0.73-10.34), and multiple myeloma (SIR 1.98; 95% CI 0.54-5.06) were also increased, not reaching statistical significance. Risk reduction was observed for malignant tumors of the gallbladder (SIR 0.36; 95% CI 0.12-0.83). CONCLUSION: In proposing recommendations for the follow-up and management of women with breast cancer, it is important to recognize their long-term predisposition to an array of secondary cancers. PMID- 11005134 TI - [Efficiency of conventional glass wool and SpermFertil columns with respect to ROS-reduction, leukocyte reduction, and CASA-generated sperm counts in semen]. AB - OBJECTIVE: Glass wool filtration has proved to be useful to separate motile from defective spermatozoa and leukocytes in semen from patients with oligozoospermia and azoospermia. With regard to that, we set out to investigate the efficiency of two widely used filter systems. MATERIAL AND METHODS: Semen from 10 patients attending the Dept. of Andrology, FSU Jena, were investigated with home-made coarse glass wool filters and SpermFertil columns regarding elimination of reactive oxygen species (ROS) and enrichment of semen with motile and progressive spermatozoa. RESULTS: Filtration with home-made glass wool columns did not affect the percentage of motile spermatozoa but increased that of progressive spermatozoa. The whole cell count was decreased by a third. Leukocytes were reduced by half and ROS by more than half. SpermFertil columns increased the percentage of both motile and progressive spermatozoa and the whole cell count was decreased severely. Leukocytes were eliminated almost completely, as were ROS. CONCLUSIONS: Our results show the higher potential of SpermFertil columns to enrich semen specimens with motile and progressive spermatozoa and to almost completely eliminate leukocytes and ROS in comparison to home-made glass wool columns. However, SpermFertil columns reduce the whole cell count to about 10% of the original cell count. PMID- 11005135 TI - [Desmoid tumor of the breast]. AB - The desmoid tumor of the breast with infiltrative and destructive growth tendencies is a rare benign lesion. Fibromatosis management favors wide local excision with clear margins, adjuvant hormonal- and radiotherapy can improve recurrence-free survival rate. We report on a patient with desmoid tumor of the breast. PMID- 11005136 TI - [Atypical process of acute disturbance of liver function with severe thrombocytopenia in the third trimester]. AB - The authors diagnosed disturbance of liver-function associated with severe thrombopenia in a pregnant woman in the third trimester. Principally, acute fatty liver of pregnancy can be characterized by existing symptoms, e.g. nausea, vomiting, epigastric pain, jaundice, hyperbilirubinemia, moderately elevated SGOT and SGPT levels, thrombopenia, leukocytosis, low fibrinogen level and disseminated intravascular coagulopathy, but hepatomegaly, purpura and petechia on lower and upper extremities, and high ALP and GGT levels during postpartum period do not confirm suspicion of this diagnosis. The present report draws attention to the difficulties of differential diagnosis of pregnancy-induced elevated liver enzymes diseases associated with low platelets, as there are several identical pathophysiological processes. Although causes and exact pathophysiology of disorders are unknown, similar symptoms during the process of diseases leave the question open whether they are different diseases or whether they are different manifestations of the same disease, and what kind of relationship exists between these diseases and preeclampsia. This case suggests careful evaluation of the whole clinical picture, moreover it is emphasized that prompt, aggressive treatment of hemostatic disturbance and the expeditious delivery can save maternal life. PMID- 11005137 TI - [Laparoscopic single patient use instruments: expensive outsourcing of product quality?]. AB - The supply of medical goods is an important critical success factor in German hospitals. One major managerial area in the procurement concerns the decision between single patient use (SPU) and multiple patient use (MPU) products. Especially laparoscopic instruments which are generally expensive are a field of interest for decision makers. Due to a lack of quantifiable factors describing the two different forms of supply alternatives with their effects on effectivity and efficiency of the procurement process and the final use are often not taken into account. Since it is expected that in the future more and more laparoscopic instruments will be needed there is a necessity for finding a concept allowing the identification of the "right" product. The Center for Hospital Management (CKM) has the aim to develop a corresponding approach but needs the help of the reader. PMID- 11005138 TI - [Electronic documentation in medicine; flexible concepts versus isolated solutions]. AB - Computer-based medical documentation so far proved advantageous especially through standardization of data entry and increased access speed. Additional benefits can be achieved through the implementation of integrated, cross-project documentation tools and their integration into the clinical work-flow, which allow data to be used for a wide variety of applications (e.g. quality management, clinical research, clinic management). The presence of incompatible documentation software often complicates the realization of these goals. Implementation of new documentation tools therefore should consider flexibility and multiple-use of data as primary design goals. In the presented paper requirements for flexible documentation tools are introduced. The Entity Attribute-Value-Model is described as a possible means of implementation. Practical experiences made with a prototype application are reported. PMID- 11005139 TI - [Interdisciplinary workshop "Hormone substitution and cardiovascular system". Stuttgart, April 13-l4, 2000]. PMID- 11005140 TI - Hereditary colorectal cancer: risk assessment and management. AB - There are at least nine major cancer susceptibility syndromes that infer an increased risk for colorectal cancer and/or colorectal polyposis; hereditary nonpolyposis colorectal cancer syndrome, Muir-Torre syndrome, Turcot syndrome, the I1307K polymorphism of the APC gene, familial adenomatous polyposis, attenuated familial adenomatous polyposis, Peutz Jeghers syndrome, juvenile polyposis, and the PTEN hamartoma tumor syndrome. As a result, the differential diagnosis of hereditary colorectal cancer can be complex. In addition, there has been a dramatic increase in the knowledge available regarding risk assessment and management of hereditary colorectal cancer syndromes. The literature was reviewed to develop this concise review of the hereditary colorectal cancer syndromes to facilitate the accurate diagnosis of each syndrome and the appropriate medical care for individuals with these diagnoses. Referral to a qualified Clinical Cancer Genetics program is appropriate if any of these syndromes is suspected and they will ensure the most up-to-date information is available to the patient, their family, and their health care professionals. PMID- 11005141 TI - Low-density lipoprotein receptor gene mutations in a Southeast Asian population with familial hypercholesterolemia. AB - The aim of this study was to detect mutations in the genes coding for the low density lipoprotein receptor and apolipoprotein B in patients of Southeast Asian origin with clinically diagnosed familial hypercholesterolemia (FH) and to relate these findings with the observed lower incidence of coronary heart disease in this part of the world. A total of 86 unrelated patients with FH were selected on clinical grounds, and complete DNA analysis of the low-density lipoprotein (LDL) receptor and apolipoprotein B (apoB) genes by DGGE and DNA-sequencing was performed. In the majority (73%) of the cohort studied, no mutations could be detected, even after extensive analysis of the LDL-receptor and apoB genes. However, the 22 patients with a mutation had significantly more xanthomas and a higher incidence of coronary heart disease and levels of low-density lipoproteins were also significantly different. There was no correlation between the type of the mutation and lipoprotein levels or clinical signs of atherosclerosis. The fact that the majority of the FH patients studied had no detectable mutation and that this group had a significant milder phenotype, suggests the presence of a third gene in the Southeast Asian population, predominantly leading to a disorder resembling a milder form of FH. A similar, but less frequent, trait has recently been described in a number of European families. PMID- 11005142 TI - Trends in the frequencies of consanguineous marriages in the Israeli Arab community. AB - To assess the trends in the frequency of consanguineous marriages in the Israeli Arab population in the last 40 years, we conducted a two-part study. For the first part, we re-analyzed data from a nationwide study carried out in 1992, and for the second part, we undertook a new survey in 1998 in four locations: Taibe, Tira, Kalansuwa and Kafr Bara. Data regarding the frequency of consanguineous marriage in these four locations for the years 1961-1985 was extracted from the original survey, and for the years 1986-1998, from new questionnaires. The frequency of consanguineous marriage was highest in the period 1961 1965 (50.6%), but by the period 1981-1985 it had decreased to 40.6%. Over the whole time span of the 1992 study, a significant decrease was observed between the periods 1961 1975 and 1976-1985 (p < 0.0001). In the four-location study, there was a significant decrease in the frequency of consanguineous marriage from 52.9%, in the period 1961-1970 to 32.8% in the period 1991-1998 (p = 0.0006). We conclude that the custom of consanguineous marriage in the studied population is still extremely high, and preventive measures should be taken to decrease its frequency and associated complications. PMID- 11005143 TI - Prenatal diagnosis of fragile X syndrome and the risk of expansion of a premutation. AB - The aim of the present study was to evaluate prospectively the dynamics of the FMR1 gene. The risk of full mutation among pregnant women and the carriers, and the risk of expansion of a premutation allele to a full mutation were estimated. We identified 89 pregnant women with an expanded FMR1 gene seeking prenatal diagnosis. Amniocentesis or chorion villus sampling (CVS) was offered and a DNA test of the FMR1 gene was carried out in such pregnancies. The overall risk of full mutation among women (N = 21) with a repeat size between 60 and 80 was 4.8% (one fetus with mosaicism), and the risk of expansion of the premutation allele to a full mutation was 14% in those offspring to whom the premutation allele was transmitted. The risk of full mutation among the carriers (N = 13) with a repeat size between 81 and 100 was 61.5% (8/13), and the risk of expansion of a premutation allele to a full mutation was 89%. Only one case fell into the category of 101-200 repeats, and expansion to a full mutation was recorded. Fetuses of full mutation mothers inherited the larger allele in 64% (14/22) of the cases. The range of 40-59 repeats was safe: there were no fetal full mutations. The risk of full mutation was also low among the subjects with a repeat size between 60 and 80, whereas the risk increased significantly after 80 repeats. Maternal premutation size was positively correlated with the risk of having a full mutation offspring. PMID- 11005144 TI - Molecular cytogenetic analysis of a duplication Xp in a female with an abnormal phenotype and random X inactivation. AB - We describe a female infant with severe abnormal phenotype with a de novo partial duplication of the short arm of the X chromosome. Chromosome painting confirmed the origin of this X duplication. Molecular cytogenetic analysis with fluorescence in situ hybridization (FISH) was performed with YAC probes, further delineating the breakpoints. The karyotype was 46, X dup(X)(p11-p21.2). Cytogenetic replication studies showed that the normal and duplicated X chromosomes were randomly inactivated in lymphocytes. In most females with structurally abnormal X chromosomes, the abnormal chromosome is inactivated and they are phenotypically apparently normal relatives of phenotypically abnormal males having dupX. Therefore, in this case, there is functional disomy of Xp11 p21.2 in the cells with an active dup(X), most likely resulting in abnormal clinical findings in the patient. PMID- 11005145 TI - Prevalence of the C282Y mutation for haemochromatosis on the Island of Majorca. AB - The C282Y mutation of the HFE gene has been reported to be present in most of the patients with hereditary haemochromatosis (HH) of Northern European ancestry. HH affects approximately 1/300 individuals, but it is not evenly distributed in the different European countries. In the present study, polymerase chain reaction (PCR) and restriction-enzyme digestion were used to analyse the frequency of the most important mutation in haemochromatosis (C282Y) in subjects from Majorca (Balearic Islands, Spain) and patients with haemochromatosis. The results were compared with other studies from Spain and Europe. A total of 420 Majorcan chromosomes were analysed and the C282Y mutation was observed at a frequency of 2.62%+/-0.8 (11 heterozygotes: eight men and three women). In the group of hereditary haemochromatosis probands, 13 out of 14 were homozygous for the C282Y mutation. In the distribution of the C282Y mutation, a north-west to south-east cline was detected, supporting the Celtic origin of this mutation. PMID- 11005146 TI - Telomere-telomere (end to end) fusion of chromosomes 7 and 22 with an interstitial deletion of chromosome 7p11.2-->p15.1: phenotypic consequences and possible mechanisms. AB - We report a rare case of a de novo end to end fusion of chromosomes 7 and 22 in conjunction with an interstitial deletion of chromosome 7p11.2p15.1 in a newborn with congenital anomalies. The proband presented for chromosome analysis with bilateral cataracts, dysmorphic facies and distal limb abnormalities. Chromosome analysis showed a 45,XY,der(22)psu dic(22;7)(p13;p22.3)del(7)(p11.2p15.1) karyotype. This short arm to short arm fusion of chromosomes 7 and 22 resulted in a pseudodicentric chromosome. The interstitial deletion in the short arm of chromosome 7 was likely a result of breakage and reunion related to instability of the dicentric chromosome. Loss of genetic material in this region of chromosome 7p has been implicated in the pathophysiology of craniosynostosis and cephalopolysyndactyly syndromes. PMID- 11005147 TI - Intrachromosomal triplications: molecular cytogenetic and clinical studies. AB - A newborn boy had meconium aspiration syndrome, hypospadias, a supernumerary digit on the left hand, hyperbilirubinemia, a fractured right clavicle, osteopenia, liver calcification, and mild pulmonary hyperplasia. Cytogenetic studies showed a chromosome 13 with additional material in 33% of the metaphases. The add(13) was considered to be a probable duplication of 13q12q22. The 13 paint probe hybridized to the add(13) from end to end. Fluorescence in situ hybridization (FISH) studies using retinoblastoma probe (RB)-1 that maps to 13q14 and D13S585 that maps to 13q32-q33 gave one signal for RB and three signals for D13S585. The pattern of the three signals from the 13q32q33 region and the G banding pattern was best explained as a triplication of 13q22q33, with an inverted middle repeat resulting in tetrasomy for this segment. Mosaicism was confirmed by FISH using a D13S585 probe on a buccal smear. Three triplications detected in our laboratory were compared 13q22q33, 15q11q13, and 2q11.2q21. FISH was critical in identifying triplications 13q22q33 and 15q11q13. The hybridization pattern also indicated an inverted middle repeat. We conclude that intrachromosomal triplications may be more prevalent than previously assumed and they probably share a common mechanism in their formation. When the G-bands do not correspond exactly to a duplication or to a tandem triplication, an important consideration is that the majority of triplications have an inverted middle repeat. Triplications can be mistaken for duplications. Therefore, in assessing duplications, FISH confirmation is recommended. PMID- 11005148 TI - Chronic myelomonocytic leukaemia in a child with constitutional partial trisomy 8 mosaicism. AB - We present a 3-year-old boy with constitutional partial trisomy 8 mosaicism (karyotype 47,XY, + del(8)(p12)/46,XY) who developed chronic myelomonocytic leukaemia and we review the few reported cases of constitutional trisomy 8 mosaicism (CT8M) associated with malignancy. This case highlights the association between CT8M and the development of malignancies, haematological malignancies in particular. PMID- 11005149 TI - Mild clinical phenotype associated with R1158X/S549R(T-->G) CFTR genotype. PMID- 11005150 TI - Genetic homogeneity of the Camurati-Engelmann disease. PMID- 11005151 TI - Risk estimates for carriers of chromosome reciprocal translocation t(4;9)(p15.2;p13). PMID- 11005152 TI - A novel closed-tube quantitative--PCR method for enumerating Porphyromonas gingivitis, Prevotella intermedia and Actinobacillus actinomycetemcomitans. AB - Enumeration of specific periodontopathogens in subgingival plaque samples has been problematic because of either lack of sensitivity, specificity or the time taken to identify the organisms. These problems can be overcome using PCR, but quantification by this technique is more difficult. We report a simple quantitative PCR method developed for enumerating Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans in clinical samples. The method relies upon inclusion of ethidium bromide in a closed-tube PCR reaction and measurement of resultant amplicons by quantifying the fluorescence generated when UV-light is passed through the walls of the amplification tube. Hence, both PCR and detection are performed in the same tube. The technique was compared with a quantitative competitive PCR and a commercial colorimetric quantitative PCR and proved to be at least as sensitive, specific and reliable for enumeration of the target bacteria. However, its speed and convenience make it particularly useful for large-scale analyses in both clinical laboratories and epidemiological studies. PMID- 11005153 TI - Salivary matrix metalloproteinase (MMP-8) levels and gelatinase (MMP-9) activities in patients with type 2 diabetes mellitus. AB - We studied the salivary levels and activities of the matrix metalloproteinases (MMP) -8 and -9 in 45 type 2 diabetic patients and 77 control subjects. The patients' mean glycosylated haemoglobin (HbA1c) was 8.7%, indicating an unsatisfactory metabolic control of the disease. The MMP levels were further related to the clinical and microbiological periodontal findings as well as to salivary flow rate and other factors. The salivary flow rate, albumin and amylase concentrations were similar in type 2 diabetic patients to those in the control group. The mean gingival and periodontal pocket indexes were higher in the diabetes group. The number of potential periodontopathogenic bacteria was lower, however, in the diabetic than in the control group. Zymography and immunoblotting revealed that the major MMPs in the type 2 diabetic patients' saliva were MMP-8 and MMP-9. Salivary MMP levels and activities in type 2 diabetic patients were in general similar to those in the control group. However, the correlation coefficients using multiple regression analysis revealed that gingival bleeding, pocket depths and HbA1c were associated with increased MMP-8 levels which, in turn, were negatively predicted by elevated plasma lipid peroxide levels in the diabetic group. Our data on salivary MMP-8 and -9 do not support the concept of generalized neutrophil dysfunction in unbalanced diabetes. Moreover, plasma lipid peroxidation levels reflecting the increased oxidative burden, which is generated mainly by triggered neutrophils, do not indicate neutrophil dysfunction due to diabetes, but may rather be related to the increased tissue damage in an uncontrolled disease. However. advanced periodontitis in type 2 diabetes seems to be related to elevated salivary MMP-8 levels which might be useful in monitoring periodontal disease in diabetes. PMID- 11005154 TI - In vitro evaluation of Er:YAG laser scaling of subgingival calculus in comparison with ultrasonic scaling. AB - The purpose of the present study was to evaluate the effectiveness of Er:YAG laser scaling and the morphological and histological changes of the laser-scaled root surface in comparison with the effectiveness and root surface changes produced by conventional ultrasonic scaling. Fifty-three periodontally involved human extracted teeth with a band of subgingival calculus were used. The teeth were divided randomly into 2 groups for laser scaling and ultrasonic scaling. Laser irradiation was performed at an energy output of 40 mJ/pulse and 10 pulses/s under water spray, with the probe tip contacted obliquely to the root surface. Ultrasonic scaling was performed at a clinically standard power setting. The time required for scaling, the scaled area and the temperature changes were determined using both methods of treatment. The features of the scaled surfaces were examined by histological and scanning electron microscope (s.e.m.) observations. The Er:YAG laser provided subgingival calculus removal on a level equivalent to that provided by the ultrasonic scaler, without major thermal elevation. Macroscopically, the laser-treated root surface was somewhat rougher than or similar to the ultrasonically scaled root. However, the efficiency of the laser scaling was lower than that of the ultrasonic scaling. In addition, histological examination revealed a thin deeply stained zone on the lased root surface, and s.e.m. analysis revealed a characteristic microroughness on the lased surface. The laser scaling provided a level of calculus removal that was similar to that provided by the ultrasonic scaling. However, the Er:YAG laser produced superficial, structural and thermal microchanges on the root cementum. PMID- 11005155 TI - Herpesviruses 6, 7 and 8 in HIV- and non-HIV-associated periodontitis. AB - Human herpesviruses, especially cytomegalovirus and Epstein Barr virus type-1, occur with higher frequency in subgingival specimens from periodontitis lesions than from healthy/gingivitis sites. Little or no information is available on the relationship between herpesvirus 6 (HHV-6), herpesvirus 7 (HHV-7) and herpesvirus 8 (HHV-8) and periodontal disease. This study determined the periodontal occurrence of HHV-6, HHV-7 and HHV-8 in 21 HIV-seropositive and 14 HIV-negative adults affected by periodontitis. Gingival biopsy specimens and paper-point samples of subgingival plaque were collected from sites showing 5 mm or more in probing depth. Nested polymerase chain reaction methodology was employed in herpesvirus identification. In the HIV-seropositive periodontitis group, 90% of gingival biopsies and 62% of subgingival plaque samples revealed at least one of the test viruses. HHV-6 occurred in 71%, HHV-7 in 67% and HHV-8 in 24% of gingival biopsies. In the HIV-negative adult periodontitis group, 43% of gingival biopsies showed at least 1 of the test viruses, with HHV-6 present in 21% and H HV-7 in 29% of gingival biopsies and with no detection of HHV-8. The combined occurrence of the 3 test herpesviruses was significantly higher in HIV seropositive than in HIV-negative adult periodontitis patients (p = 0.008). The human periodontium might constitute a site of infection or reservoir for HHV-6, 7, -8. PMID- 11005156 TI - Glucocorticoid receptor antagonist RU 486 treatment reduces periodontitis in Fischer 344 rats. AB - Inappropriate hypothalamic pituitary adrenal (HPA) axis regulation of immune responses to bacterial challenges has been found to play an important role in infections and inflammatory disease susceptibility and progression. In the present study we investigated the tissue effects of experimental periodontitis in Fischer 344 rats, which were subcutaneously (s.c.) injected with 20 mg/kg of the glucocorticoid receptor antagonist and active antiglucocorticoid agent RU 486 every second day over a period of 14 d. Periodontitis was induced by placing a bacterial plaque retentive silk ligature in the gingival sulcus around the neck of maxillary right 2nd molar teeth 1 d after the first injection in 10 RU 486 treated and 10 vehicle (1,2-propanediol)-treated control animals. The contralateral maxillary left 2nd molars served as internal control teeth for naturally occurring periodontitis. Disease progression was evaluated radiographically and histometrically. The average level of corticosterone in blood at sacrifice was significantly lower in the RU 486-treated animals as compared to controls. The experimental animals also developed significantly less periodontal breakdown at both experimental and control teeth compared to the vehicle-treated control animals. The results support our recent findings showing that HPA hyper-reactivity, either genetically determined or experimentally induced, stimulates periodontal disease susceptibility. These findings suggest that central nervous regulation of inflammatory responses to dental plaque microorganisms in the gums may modulate periodontal disease susceptibility and progression. PMID- 11005157 TI - Cytostatic action of enamel matrix derivative (EMDOGAIN) on human oral squamous cell carcinoma-derived SCC25 epithelial cells. AB - During surgical treatment of periodontal disease, enamel matrix derivative (EMD) is topically applied as a substitute for extracellular matrix in order to facilitate regeneration of damaged periodontal tissue. However, the mechanism for EMD action is poorly understood. We have now examined the effects of EMD on the proliferation of oral epithelial (SCC25) cells in vitro. After 3 days of treatments, EMD (25 100 microg/ml) dose-dependently inhibited cell division and concomitantly arrested cell cycle at the G1 phase. Prior to this inhibition, EMD significantly up-regulated p21WAF1/cip1, a cyclin-dependent kinase inhibitor, induced G1-arrest, and inhibited DNA synthesis. In addition, EMD down-regulated expression of cytokeratin-18 (CK18) protein, which was most due to decreased production, but less to increased degradation. However, EMD did not discernibly increase the number of apoptotic cells over 8 days of treatment. These findings indicate (1) that EMD acts as a cytostatic agent, rather than a cytotoxic agent, on epithelial cells, and (2) that this anti-proliferative action is probably due to p21WAF1/cip1-mediated G1-arrest. Furthermore, our in vitro cellular data clearly verify and provide an explanation for the clinical observation that EMD application suppresses the down-growth of junctional epithelium onto dental root surfaces, a process that frequently interferes with the formation of new connective tissue attachments. PMID- 11005158 TI - Altered expression level of calbindin D28k in the periodontal ligament of rat molar in response to changes in occlusal force. AB - The present immunohistochemical study was designed to investigate the alteration in the expression level of calbindin D28k in the periodontal ligament of the rat molar in response to changes in occlusal force to clarify the physiological role(s) of this protein in the ligament. In normal periodontal ligament of the lower first molar, immunoreactivity for calbindin D28k was found in the spindle shaped cells, presumably fibroblasts, at the alveolar portion of the ligament at the distal side of the mesial root and mesial side of the distal root. Following the overload of occlusal force to the upper first molar by bite-raising, the number and immunoreactivity of the positive cells in the periodontal ligament of the lower first molar increased gradually. A more significant increase was detected at 7 d following the bite-raising compared to the normal animals. When occlusal force was removed by the extraction of the upper first molar, the expression level of calbindin D28k in the periodontal ligament of the lower first molar rapidly decreased, however a subsequent gradual increase was recognized. Statistical analysis of the spatial immunoreactivity of calbindin D28k in the periodontal ligament was performed and showed statistically significant differences. The present results suggest that calbindin D28k may play important roles in the homeostasis and cytoprotection of the periodontal fibroblasts against occlusal force. PMID- 11005160 TI - Floating the data acquisition chains to improve the quality of measurements. AB - Guarding sensors and measuring equipment is a smart operation which decouples the apparatus or systems from common mode electrical disturbances. In this context, the floating techniques are revisited and the usual measurement problem in noisy environment is outlined. It is shown that the use of differential inputs to pickup sensor continuous signals, as well as simple shielding and grounding techniques lead to increase the quality of data acquisition systems. At the other size, optical couplers allow to pass the digital version of data to an external bus or to a network by preserving the apparatus floating features. PMID- 11005159 TI - Improvement of macromolecular clearance via lymph flow in hamster gingiva by topical warming and massage. AB - The lymphatic system is very important for macromolecular clearance in various tissues, especially in the gingiva. However, the kinetics of macromolecular clearance via the lymph flow in the gingiva are poorly understood. The aim of this study was to investigate whether thermal or mechanical stimulation affects macromolecular clearance via the lymph flow in the gingiva. Carbon black suspension was injected into the mandibular gingiva of anesthetized hamsters and its drainage into cervical lymph nodes was examined. Clearance of 14C-methylated bovine albumin and tritiated water from the gingiva and their drainage into submandibular lymph nodes and blood was quantified. The effect of topical warming or massage on clearance of 14C-methylated albumin from the gingiva during a 15 min period was examined. In addition, the influence of neurochemical antagonists on the stimulatory effect of topical warming on albumin clearance was investigated. Submandibular lymph nodes were clearly delineated by carbon black 10 min after the injection. More radiolabeled albumin appeared in submandibular lymph nodes than in serum, while more tritiated water appeared in serum. Topical warming (45 degrees C, 2 min) and warming plus massage (with a silicon rubber brush, 20 s) decreased the radiolabeled albumin in the gingiva 15 min after the injection. There was less radiolabeled albumin in the gingiva after gingival warming plus massage than after warming. Previous injection of HOE140 or propranolol into the gingiva diminished the stimulatory effect of topical warming on albumin clearance. It was concluded that topical warming plus massage improves macromolecular clearance via the lymph flow in hamster gingiva. PMID- 11005161 TI - Pattern classification by a neurofuzzy network: application to vibration monitoring. AB - An innovative neurofuzzy network is proposed herein for pattern classification applications, specifically for vibration monitoring. A fuzzy set interpretation is incorporated into the network design to handle imprecise information. A neural network architecture is used to automatically deduce fuzzy if-then rules based on a hybrid supervised learning scheme. The neurofuzzy classifier proposed is equipped with a one-pass, on-line, and incremental learning algorithm. This network can be considered a self-organized classifier with the ability to adaptively learn new information without forgetting old knowledge. The classification performance of the proposed neurofuzzy network is validated on the Fisher's Iris data, which is a well-known benchmark data set. For the generalization capability, the neurofuzzy network can achieve 97.33% correct classification. In addition, to demonstrate the efficiency and effectiveness of the proposed neurofuzzy paradigm, numerical simulations have been performed using the Westland data set. The Westland data set consists of vibration data collected from a US Navy CH-46E helicopter test stand. Using a simple fast Fourier transform technique for feature extraction, the proposed neurofuzzy network has shown promising results. Using various torque levels for training and testing, the network achieved 100% correct classification. PMID- 11005162 TI - The development of reactor coolant pump vibration monitoring and a diagnostic system in the nuclear power plant. AB - Monitoring abnormalities within reactor coolant pumps due to vibration, we introduce a Wigner distribution (WD) to a vibration monitoring system for analyzing vibration signals, and develop an on-line diagnostic method using the neural network employing back propagation algorithm. A rotor kit is used to simulate the abnormal conditions in the pump such as bearing rubbing, shaft bending, misalignment, etc. The data collected from the kit are analyzed using both a conventional system based on the Fourier transform (FT) and the software developed with the WD in this study, and they are compared. The study shows that spectrum represented by WD makes it easy to analyze the vibration signals of a rotating machine and the developed diagnostic method will help the operators grasp the cause of vibrations. PMID- 11005163 TI - PID gain scheduling using fuzzy logic. AB - A simple, yet robust and stable alternative to proportional, integral, derivative (PID) gain scheduling is developed using fuzzy logic. This fuzzy gain scheduling allows simple online duplication of PID control and the online improvement of PID control performance. The method is demonstrated with a physical model where PID control performance is improved to levels comparable to model predictive control. The fuzzy formulation is uniquely characterized by; (i) one fuzzy input variable involving the PID manipulated variable, (ii) two parameters to be tuned, while previously tuned PID parameters are retained, and (iii) a gain scheduling differential equation which relates the fuzzy and conventional PID manipulated variables and enables fuzzy gain scheduling. PMID- 11005164 TI - Intelligent control of a cryogenic cooling plant based on blackboard system architecture. AB - Intelligent system techniques have been rapidly assimilating into process control engineering, with many applications reported in the last decade. Intelligent control is bringing a new perspective as well as new challenges to process control. In this paper, a software architecture for a Blackboard for Integrated Intelligent Control Systems (BIICS) is described. The system is designed to simultaneously support multiple heterogeneous intelligent methodologies, such as neural networks. expert systems, fuzzy logic, neural networks and genetic algorithms. It will be shown how such methodologies can be readily assimilated into the software architecture. The BIICS system represents a multi-purpose platform for design and simulation of intelligent control paradigms for different kinds of processes. Currently the system utilizes intelligent control techniques (neuro-fuzzy and genetic optimization) for controlling a cryogenic plant used for superconductor testing at temperatures below 100 K. PMID- 11005165 TI - Soft-testing of industrial control systems programmed in IEC 1131-3 languages. AB - This paper presents a technique and the means for testing PLC-based control software outside the actual plant environment with the purpose of increasing the confidence level on the compliance of the software to functional and temporal requirements. The need to obtain a high confidence level on the correct software operation arises from the fact that in most of the cases it is quite dangerous and expensive to test unproved PLC operation by linking it with the actual facilities that it is going to control. The proposed technique relies on a combined simulation of the controlled plant and the PLC system and an analysis of the plant responses. The PLC system simulation imitates the way software is executed on a PLC that is programmed in the languages of the IEC1131-3 standard. It is based on the programming model of the IEC standard and analytical formulae for estimating the program runtime. The simulation of the plant is based on a discrete convolution model that is solved at the same rate with the rate determined by the control algorithms. A tool realizing these concepts has been developed and its use in testing the control software of three critical outputs of a distillation column is demonstrated. PMID- 11005166 TI - Expanding the applicability of ISA TR84.02 in the field. AB - The ANSI/ISA S84.01 standard was released in 1996. The companion technical report TR84.02 is in the process of being completed. The latter document is intended to provide the methodology to implement the safety performance requirements of the standard for the safety system. In this document, three techniques are presented; these being Simplified Equations, Fault Tree Analysis and Markov Modeling. Of the three, only the Simplified Equations approach would reasonably be utilized in the field by plant personnel. The Simplified Equations provided as part of TR84.02 comprehend common cause failures, systematic failures, and second failure prior to repair scenarios. They do not however comprehend the use of redundant field devices which are dissimilar, and as such have different failure rates. This situation is quite common in practice, and simple to manage using enhanced equations for the computation of PFDavg. A set of these equations for typical redundant architectures in the field, and several examples of their applications in safety loop analysis are derived and presented in this paper. PMID- 11005167 TI - A method to prevent excessive numbers of Markov states in Markov models for quantitative safety and reliability assessment. AB - This paper presents a method that will drastically reduce the calculation effort required to obtain quantitative safety and reliability assessments to determine safety integrity levels for applications in the process industry. The method described combines all benefits of Markov modeling with the practical benefits of reliability block diagrams. PMID- 11005168 TI - Consecutive matching striation criteria: a general critique AB - In the forensic science of firearms and toolmark identification, examiners traditionally have drawn conclusions of identity from subjective criteria. This paper critically explores the general validity of one proposed objective-criteria regime--that of counting consecutive matching striations on fired bullets. Practical considerations are discussed, as well as theoretical ones, with both discussions viewed from the perspective of Bayesian logic. It is concluded that drawbacks exist for this particular objective-criteria regime, but that research and logical analysis should continue. PMID- 11005169 TI - Identification of heroin in street doses using 1D-TOCSY nuclear magnetic resonance AB - Heroin street doses are complex mixtures commonly analyzed in forensic laboratories. Identification of the illicit substance in these street doses is among the primary analytical tasks of a forensic laboratory. We demonstrate that the one-dimensional ID-TOCSY NMR experiment permits identification of heroin in standard mixtures containing up to ten or more different components. This method produces an easily-identified and effective "fingerprint" for heroin within a mixture of other substances. The method has been successfully tested as a tool for identification of heroin in street doses from police casework in Israel. This NMR technique is robust and quick (a measurement can be carried out in 10-15 min), and it does not require any preliminary physical or chemical treatments of the sample to be examined, due to the effective spectroscopic "filtering" of the interfering components. The ID-TOCSY NMR method can potentially be used in combination with additional analytical methods as a routine tool in forensic laboratories to positively identify heroin for court purposes. PMID- 11005170 TI - The petroleum-laced background AB - Using passive headspace concentration (ASTM E-1412) and gas chromatographic/mass spectrometric (GC/MS) analysis as described in ASTM E-1618, the authors have studied the volatile components detectable in several kinds of otherwise uncontaminated substrata, including clothing, shoes, household products, building materials, paper products, cardboard, and adhesives. Due to the use of petroleum derived liquids in the manufacture of these materials, it is frequently possible to detect the liquids, even when the products are several years old. These results point out the need for the use of comparison samples whenever possible. PMID- 11005171 TI - A novel method for forensic DNA investigations: repeat-type sequence analysis of tandemly repeated mtDNA in domestic dogs. AB - A highly variable and heteroplasmic tandem repeat region situated in the mitochondrial mt DNA control region (CR) in domestic dogs and wolves was studied to evaluate its suitability as a forensic genetic marker for analysis of single hairs. The tandem repeat array is composed of three 10-bp repeat types that are distributed so that a secondary DNA sequence is formed. Thus, the region presents two levels of variation: variation in the number of repeats and variation in the secondary DNA sequence of repeat types. Two analysis methods were therefore tested; fragment length analysis and analysis of the sequence of repeat types. Fragment analysis produced unique profiles that could be used to discriminate between blood samples from maternally closely related individuals. However, different hairs from one individual did not have the same fragment profile, and the method is, therefore, not suitable for analysis of single hairs. In contrast, analysis of the repeat type sequences (array types) is highly informative. When different hairs from one individual were studied, identical array types were found. The repeat-type sequence variation was studied among individuals having identical nonrepetitive CR mtDNA sequence variants. Seven, six, and two individuals, representing three different sequence variants, respectively, were analyzed. All these individuals had different array types, which implies a very high genetic variation between individuals in this region. The analysis method considerably improves the exclusion capacity of mtDNA analysis of domestic dogs compared with sequence analysis of non-repetitive DNA. PMID- 11005172 TI - Improved method for shooting distance determination. Part 2--bullet holes in objects that cannot be processed in the laboratory AB - An improved method for firing distance determination on exhibits that cannot be processed in the laboratory such as cars, doors, windows, or furniture is described. The novel part of the method includes transfer of total nitrite (nitrite ions and smokeless powder residues) from the target to an adhesive lifter. After the transfer, vaporous lead and copper deposits around the bullet entrance hole are visualized on the target by sodium rhodizonate and rubeanic acid, respectively. The Modified Griess Test is carried out after alkaline hydrolysis of the smokeless powder residues on the adhesive lifter. PMID- 11005173 TI - Analysis of genotype frequencies and interlocus association for the PM, DQA1, and D1S80 loci in four populations. AB - Allele frequencies of the LDLR, HBGG, GYPA, D7S8, GC, DQA1, and D1S80 loci are presented and genotypes are analyzed for each of four ethnic groups: African Americans (n = 200), US Caucasians (n = 200), US Hispanics (n = 200), and Japanese (n = 89). Hardy-Weinberg genotypic proportions were observed in all but two of the 28 population-locus tests undertaken. Those two instances are attributable to type I statistical error. Gametic equilibrium among loci is an assumption invoked for application of the product rule to utilize the discriminatory power from two or more loci simultaneously. Two statistical methods, a genotype matching statistic and log-linear modeling, were used to evaluate gametic disequilibrium. The match statistic, comparing observed to expected likelihood of genotypic identity for seven loci among pairs of individuals within the database, revealed only one statistically significant deviation among 20 tests. As expected, the probability of match was generally lowest in the test on all ethnic groups combined, indicating that allele frequencies differ among ethnic groups for some of the loci. This was confirmed with the statistic theta to measure ethnic stratification, in which about 0.10 of the genetic variation is apportioned among the four ethnic groups for four of the structural loci (LDLR, HBGG, GC, and DQA1), while for GYPA, D7S8, and D1S80, variation is more uniformly distributed among ethnic groups. Log-linear modeling was also applied to the five PM loci. The most parsimonious log-linear model included only three higher order terms: the two-way interactions of three of the PM loci with ethnic group. These three instances (LDLR, HBGG, and GC) indicated differences in allele frequencies between ethnic groups. No two or higher way interaction (disequilibrium) was observed among loci. In summary, the assumptions of Hardy-Weinberg and gametic equilibrium that facilitate the use of the five PM loci, DQA1 and D1S80 in forensic applications are consistent with the allele and genotype frequencies observed in these populations. PMID- 11005174 TI - A rare mutation in the amelogenin gene and its potential investigative ramifications. AB - Over the past few years, the Australian forensic science community has adopted a common methodology and technology in the application of DNA profiling for investigative and forensic purposes. The ultimate objective of this initiative is the establishment of a national DNA database similar to that used in the UK. An integral part of this methodology is the use of "Profiler Plus," a nonaplex of STRs combined with amelogenin, a locus utilized for sex determination. This paper reports the results from a case where a mutation in the annealing region of the amelogenin primers appears to have resulted in the failure to amplify the amelogenin Y-homolog from a phenotypically normal male. The result was confirmed using two different primer sets that amplify different regions of the amelogenin gene. This situation suggests that the genetic determination of sex based on the amelogenin sequences from specimens of unknown origin, such as crime scene samples, should not be considered infallible. PMID- 11005175 TI - Hate crimes on the internet. AB - The Internet serves as a channel for electronic communication on an international level. While communication on the Internet has grown exponentially, the proliferation of crimes in cyberspace has become rampant. Hate crimes, in particular, have become increasingly prevalent on the Internet. In this past decade, the United States government has taken significant measures to combat the proliferation of hate crimes. This paper reports six cases of "cyberhate" crimes and emphasizes pertinent legal issues surrounding them. Current modes of intervention are discussed, ranging from local to national levels. The forensic psychiatrist may undertake a challenging role in the interpretation of the hateful criminal mind at the interface of psychiatry and the law. PMID- 11005176 TI - Scanning electron microscopic analysis of skin resolution as an aid in identifying trauma in forensic investigations. AB - The forensic investigator is frequently confronted with cases that present with wounds and blunt force trauma. Presently, the forensic investigator depends upon previous experience and further investigative deduction of the crime scene to analyze these injuries. Although not readily apparent to the naked eye, many skin tissue injuries can be visualized with scanning electron microscopy (SEM). This study was designed to establish skin trauma resolution using SEM in various skin preparations. Tissue trauma was induced on leather, preserved skin, fresh skin, and living skin using dies of varying thread size. Calibrated pressure forces in pounds per square inch (psi) were applied and impressions made using vinyl polysiloxane. Positive replicas of the tissues were prepared for SEM using isocyanate resin. After sputter coating the cast with 35 nm of gold-palladium, electron micrographs were generated using a Jeol JSM-5310LV scanning electron microscope. To establish resolution, thread widths of 52, 104, and 208 threads per inch (tpi) and trauma forces of 150, 200, and 250 psi were used to produce the impressions. Microgrooves that were identified on the die threads were analyzed. The optimum pressure for resolution studies was 150 psi using the 52 tpi die on the leather sample (4.67 +/- 0.88 microm, p = 0.046 and 0.025, respectively, by ANOVA). The resolution was compared to that of leather using preserved, fresh, and living skin. The resolution in preserved and fresh skin was less than for leather (9.00 +/- 1.73 and 10.5 +/- 4.5 versus 4.67 +/- 0.88 microm, p = 0.09 and p = 0.20, respectively). Living skin resolution was 3 microm at 52 tpi and 100 psi. Various implements of blunt force trauma were also examined using the leather sample. Time after trauma resolution was examined at 0 (3 microm), 5 (6 microm), 10 (8 microm), and 20 (9 microm) min in living tissue. A comparison between the microgrooves on the die replicas and the tissue trauma impressions revealed striking agreement for both linearity and resolution. Analysis of the microgrooves suggests that discrete morphological characteristics are seen in skin tissue traumas. This method could expand the tools available for the forensic investigation of blunt force trauma. PMID- 11005177 TI - Time trends in a study of 440 mass slayings/rampages occurring in public places. AB - We looked for variations in the timing of mass slayings/rampages (MS/R) committed in public places by solitary perpetrators. News reports on 440 MS/R from 1920 to 1996 yielded 379 onset times which were examined for temporal patterns. There was a dramatic increase in the number of MS/R from 1966 to 1995. We observed a significant monthly variation with peaks in July-August and December, and a bimodal daily rhythm for MS/R onset with a major peak at 11:00 a.m. and a minor peak at 8:00 p.m. Observed time trends for MS/R could help develop protective programs to reduce stress and violence in public places. PMID- 11005178 TI - Elimination of 7-aminoflunitrazepam and flunitrazepam in urine after a single dose of Rohypnol. AB - The hypnotic benzodiazepine flunitrazepam (Rohypnol) has been identified as the drug of choice for the purposes of "drugging" unsuspecting victims and raping them while they are under the influence of this substance. The objective of this paper was to study elimination of flunitrazepam and 7-aminoflunitrazepam in urine collected from ten healthy volunteers who received a single 2 mg oral dose of Rohypnol, to determine how long after drug administration 7-aminoflunitrazepam can be detected. A highly sensitive NCI-GC-MS method for the simultaneous quantitation of flunitrazepam (LOQ 100 pg/mL) and 7-aminoflunitrazepam (LOQ 10 pg/mL) in urine was developed. All samples were screened for benzodiazepines using optimized micro-plate enzyme immunoassay. The highest concentrations of 7 aminoflunitrazepam (70-518 ng/mL) and flunitrazepam (0.7-2.8 ng/mL) in urine were observed 6 h after drug administration in nine subjects and after 24 h in one subject. In six subjects 7-aminoflunitrazepam was detected up to 14 days after flunitrazepam administration, in one subject up to 21 days and in three subjects up to 28 days. In urine samples collected from six volunteers, flunitrazepam was detected three days after Rohypnol intake, in three subjects 24 h, and in one subject 5 days later. Benzodiazepine micro-plate enzyme immunoassay kit allowed the detection of flunitrazepam and metabolities 5 to 21 days after drug administration. PMID- 11005179 TI - Regional distribution of cocaine in postmortem brain of chronic human cocaine users. AB - We measured concentrations of cocaine and its major metabolites (benzoylecgonine, ecgonine methylester, norcocaine, and cocaethylene) in 15 autopsied brain regions of 14 human chronic cocaine users. Only slight differences were observed in concentrations of cocaine and its metabolites amongst the examined brain areas. Although it is likely that some postmortem redistribution of the drug must have occurred, our data are consistent with the possibility that behaviorally relevant doses of cocaine are widely distributed throughout the brain of humans who use the drug on a chronic basis. Consideration should therefore be given to the possible pharmacological and toxicological actions of cocaine in both striatal and extra-striatal brain areas in human users of the drug. PMID- 11005181 TI - A study on the use of photopolymer reflex plate in the forgery of Chinese seals AB - This study compares the identifying characteristics between the impressions made with original Chinese seals and photopolymer replicate seals, and between the impressions made with two photopolymer replicate seals that originate from one single negative. Various seal materials common to the Chinese community such as cow's horns and ivory are employed. Other than the common factors such as different incident angle upon imprinting or different degree of inking that constitute the natural variation in impressions, the physical changes of the seal materials with temperature will be considered. This study concludes that photopolymer replicate seal impressions can be differentiated from the originals in respect to print density and stroke morphology, but it is very difficult to differentiate between two photopolymer replicate seal impressions with the same origin. This study also proposes a simple method for the examination of forged Chinese seal impressions. PMID- 11005180 TI - Strontium isotope composition of skeletal material can determine the birth place and geographic mobility of humans and animals. AB - The Sr isotope composition measured in skeletal elements (e.g., bone, teeth, or antlers) can be used to infer the geographic region that an animal or human inhabited, because different regions tend to have distinct Sr isotope compositions, and natural variations in the relative abundance of Sr isotopes are not changed as Sr is processed through the food chain. Therefore, an organism that ingests Sr from one region can have a Sr isotope composition that is different than that of an organism that ingests Sr from another region. The Sr isotope composition of skeletal elements is a reflection of the concentration weighted average of dietary Sr that was ingested while that skeletal element was produced. Because different skeletal elements grow and exchange Sr at different stages during the life times of organisms, Sr isotope analysis of different skeletal elements can be used to infer changes in geographic location at different stages in an organism's life. The Sr isotope composition measured in human teeth will reflect the average Sr isotope composition that was ingested as a child, due to the immobile nature of Sr and Ca in teeth after formation, whereas the Sr isotope composition of bone will reflect the average isotopic composition over the last ten years of life, due to continuous biological processing of Sr and Ca in bone. Inferring the average isotopic composition of dietary Sr is best done by analyzing skeletal fragments from control groups, which might be animals that have the same feeding habits as the animal in question, or, in the case of humans, analysis of close family relatives. In cases where it is not possible to construct a Sr isotope database from control groups, it becomes necessary to estimate the isotopic composition of dietary Sr based on geologic principles. We present three case studies from our research that illustrate a range of approaches: (1) results from a criminal case where a deer was illegally harvested and the location of the deer was important to establish, (2) a pilot study of commingled human remains from a burial in Vietnam, associated with the Vietnam Conflict, and (3) a study of 13th and 14th century migration of peo ple from an archeological site in the Southwest United States. PMID- 11005182 TI - Age estimation from sternal ends of ribs by phase analysis in South African Blacks. AB - The sternal ends of ribs are used in age determination of unknown adult remains. Standards for American populations have been described and tested. The method described by Iscan et al. is reviewed and compared with other age markers of the human skeleton. Three hundred and thirty-nine (265 male, 74 female) sternal ends of right fourth ribs, belonging to black individuals deceased in the Gauteng Province of South Africa, were collected during 1994, 1995, and 1996. Three South African investigators applied the existing method and found the repeatability among them acceptable, although the method was less accurate to predict age in this population. New phases with adjusted criteria and age ranges were developed specifically for the South African black population. A tendency toward delayed maturation was found, as well as a diversion of the appearance of female ribs perimenopausally. Future studies involving more individuals in the older age ranges, and females of all ages, could broaden the representativeness of these phases. PMID- 11005183 TI - Allele and genotype frequencies for D1S80 and 3'APOB in Recanati, Central Italy. AB - The VNTR 3'APOB and D1S80 loci were studied in a sample of 179 individuals living in the Recanati Area (Central Italy). For 3'APOB, we found 34 genotypes and 11 alleles. The system was in Hardy-Weinberg equilibrium. The observed and expected heterozygosity were 0.788 and 0.798 respectively. The discrimination power was 0.96, the a-priori paternity exclusion power was 0.619 and the polymorphism information content was 0.773. For D1S80, we found 45 genotypes and 18 alleles. The system deviated significantly from Hardy-Weinberg equilibrium. The observed and expected heterozygosity were 0.696 and 0.790 respectively. The discrimination power was 0.96, the a-priori paternity exclusion power was 0.617 and PIC was 0.767. The Recanati sample was compared with the general Italian frequencies for the 3'APOB locus. A difference of borderline significance was detected (P = 0.04). For D1S80, the sample was compared with a sample from Southern Italy and no significant difference was detected. PMID- 11005184 TI - Population frequency distribution of HumF13A01, HumFXIIIB, and HumLIPOL loci in the Basque Country (Northern Spain). AB - A population study of unrelated individuals from the Basque Country (Northern Spain) was carried out using the GenePrint STR System. The PCR products were separated on denaturing polyacrylamide gels and visualized by silver staining. Three tetrameric loci were evaluated: HumF13A01, HumFXIIIB, and HumLIPOL. All loci fit Hardy-Weinberg expectations, and independence of alleles was found between these STR loci. A comparison with other population groups indicated allele frequencies are well conserved in Caucasians, but differ from other racial groups. The calculated parameters a priori probability of exclusion (Pex) and "power of discrimination" (PD) show how informative these loci are for the determination of identity and relatedness of individuals. PMID- 11005185 TI - Loss of heterozygosity detected in a short tandem repeat (STR) locus commonly used for human DNA identification. AB - Short tandem repeat (STR) markers are commonly used in basic genetic research and in human identification testing. Clinically, STRs can be used to study genetic alterations in tumors. A genetic deletion common to many types of cancer is referred to as the loss of heterozygosity (LOH). Numerous examples of LOH in cancer have been described and some have been mapped to areas located in close proximity to markers employed in human identity testing. Despite this fact, LOH has rarely been observed for STR loci commonly employed in forensic testing. Recently, for medico-legal purposes, we were asked to determine whether a tissue biopsy originated from a particular individual. For a reference source we assessed two specimens, one from normal tissue and one from cancerous tissue. When both reference specimens were used to generate DNA profiles, we observed LOH at one STR locus, D13S317. As demonstrated in other cancers only the cancerous biopsy demonstrated LOH. The forensic community should be cognizant of these unusual circumstances because, as identification of human DNA continues to be used more extensively, certain instances will arise in which reference material will not be readily available. In these situations, archived specimens may be employed as a reference source. Clinical specimens such as tissue biopsies should be used with caution if they have not been confirmed to contain normal tissue. PMID- 11005186 TI - The presumptive reagent fluorescein for detection of dilute bloodstains and subsequent STR typing of recovered DNA. AB - A presumptive reagent for dilute blood detection other than luminol is fluorescein. The sensitivity of fluorescein approaches the sensitivity of detection levels of luminol. The fluorescein detection method offers the advantages of working in a lighted environment, and the reaction persists longer than luminol. A series of diluted bloodstains, ranging from neat to 1:1,000,000, was placed on a variety of substrates. Three sets were made per substrate. One set was exposed to fluorescein, one set was exposed to luminol, and one set served as an uncontaminated control. The fluorescein signal persisted longer than luminol. However, background staining for fluorescein was observed on some substrates within 30 s to 1 min, and no background staining was observed for luminol. Stains on non-absorbent surfaces were detectable at 1:100,000 dilutions, and stains on absorbent surfaces were detectable usually at no more than 1:100. The sensitivity of detection of fluorescein was comparable to that of luminol in this study. In all cases, where sufficient DNA was recovered, typeable results at all 13 core CODIS STR loci were obtained from treated bloodstains and controls. The results from STR typing indicate that there was no evidence of DNA degradation. PMID- 11005187 TI - RFLP band size standards: NIST standard reference material 2390. AB - The procedural standard for DNA profiling developed by the U.S. advisory board on DNA quality assurance methods mandates annual confirmation of forensic DNA measurement systems against an appropriate reference material supplied by or traceable to the National Institute of Standards and Technology (NIST). NIST Standard Reference Material (SRM) 2390 is a suitable and appropriate standard for HaeIII restriction enzyme-based restriction fragment length polymorphism (RFLP) profiling systems. Originally issued in 1992, an among-laboratory SRM 2390 recertification study was initiated in 1997. Using data provided by the 20 state, local, or commercial forensic laboratory participants, quantitative band sizes values (expected mean values and associated bivariate tolerance intervals) are established for two different-source DNAs (female cell line K562 and healthy male "TAW") for genetic loci D1S7, D2S44, D4S139, D5S110, D1OS28, and D17S79. Methods for validating an RFLP measurement system, validating a control material or other secondary standard, and for tracing a particular set of RFLP measurements to NIST SRM 2390 are described in detail. PMID- 11005188 TI - RFLP band size standards: cell line K562 values from 1991 to 1997 proficiency studies. AB - Cell line K562 is the defacto forensic control material for forensic restriction fragment length polymorphism (RFLP) DNA profiling in the U.S. Fifty-one proficiency tests conducted from 1991 through 1997 enable a detailed description of RFLP measurement performance during this period. Sufficient data are available to define reference distributions for all commonly utilized and many less commonly reported genetic loci, for both HaeIII- and HinfI-based RFLP systems. The average measured size of HaeIII locus D1S7 and D5S110 bands has varied slightly over time; while relatively small, these temporal changes add to the overall interlaboratory measurement uncertainty. The characteristic standard deviation for HinfI RFLP system measurements has a nearly identical dependence on expected band size as does the standard deviation for HaeIII measurements. The ellipsoidal distance, K, is suggested for use as an RFLP data quality metric; the critical threshold value that on average excludes 1% of plausibly valid proficiency data for a given polymorphic locus is K1% = 14.2. PMID- 11005189 TI - The identification of 2-chloro-4,5-methylenedioxymethylamphetamine in an illicit drug seizure AB - This work outlines the unequivocal identification of the "ecstasy" analog, 2 chloro-4,5-methylenedioxymethylamphetamine, using combined gas chromatography/mass spectroscopy (GC/MS) and proton magnetic resonance spectroscopy (1H-NMR). This compound was identified along with 3,4 methylenedioxymethylamphetamine (MDMA) in an illicit tablet seizure, which included 26 off-white tablets. PMID- 11005190 TI - The effects of microwave irradiation on occluded solvents in illicitly produced cocaine hydrochloride AB - The current clandestine methodology for the manufacture of illicit cocaine hydrochloride utilizes microwave heating in order to dry the finished product. This study addresses the effects this step has on the occluded solvents present in newly prepared cocaine hydrochloride. Nine 1-kilogram-sized batches of cocaine hydrochloride were prepared from cocaine base using a variety of solvents or solvent mixtures commonly utilized in clandestine laboratories, pressed into bricks, and submitted to microwave heating. Residual solvents were qualitatively and quantitatively monitored before, during, and following the microwaving step by static headspace-gas chromatography-mass spectrometry. All solvents used in the conversion process were easily detected in the bricks even after extensive irradiation, confirming that occluded solvents are extremely resistant to removal by microwave heating. Qualitative and quantitative data corresponding to the residual solvents in the prepared cocaine hydrochloride bricks are presented. PMID- 11005191 TI - Analysis of biofluids for flunitrazepam and metabolites by electrospray liquid chromatography/mass spectrometry. AB - A rapid and sensitive liquid chromatography/electrospray ionization mass spectrometry (LC/MS) procedure has been developed for the analysis of biofluids containing flunitrazepam and its metabolites. Specimens were spiked with deuterated analogs of the analytes. Urine specimens were enzymatically hydrolyzed and blood specimens were untreated. Extractions were carried out using CleanScreen DAU SPE cartridges. The drugs were separated on a C18 column using a methanol-water-ammonium hydroxide (60:40: 0.03 v/v) mobile phase. After determination of base peaks using full scan mass spectrometry, the mass spectrometry method was optimized to operate in selected-ion monitoring (SIM) mode for the base peak of each analyte. Positive findings were confirmed by LC/MS/MS using the same mobile phase and column. This analytical procedure allows for the detection of low levels of flunitrazepam and metabolites in biofluids. It is useful for ascertaining the role of flunitrazepam in cases of drug-facilitated sexual assault. PMID- 11005192 TI - Parentage testing on blood crusts from firearms projectiles by DNA typing settles and insurance fraud case. AB - We describe a case of a fraudulent insurance claim. The family of an adult white male (DLF) notified the police of their son's disappearance. After a few weeks, a corpse that presented characteristics similar to those of the DLF was found in advanced stages of decay and was identified by the family as being DLF. The family then filed a claim for the life insurance that DLF had taken out just before he disappeared. Suspicions were raised about the identification of the corpse, because it had been done only visually, and because the insurance policy had been taken out just prior to DLF's disappearance. The insurance company requested a postmortem examination for identification. As the corpse had been cremated immediately after identification by the family, the biological material that was encrusted on the two projectiles removed from the body was used for analysis. The blood crusts provided enough genomic DNA for us to carry out PCR base typing of HLA-DQA1, D1S80, HUMCSF1PO, HUMTPOX, HUMTH01, D3S1744, D12S1090, D18S849, and amelogenin. Results from all loci typing from the corpse presumed to be that of DLF were then compared with that of his alleged biological parents, revealing genetic incompatibility. PMID- 11005193 TI - A cause celebre: the so-called "ballpoint murder". AB - Transorbital intracranial injuries are uncommon but classic ophthalmologic traumas. This report describes a case of a woman who was found dead. Postmortem examination revealed a Bic ballpoint which had penetrated her head through her right eye. Detective forces believed a murder to be the most likely cause; however, medical expert consultants indicated that a tragic accident was more likely. The case and the results of crossbow test-firing on human cadavers are presented. PMID- 11005194 TI - Acute cerebellar hemorrhage in a patient with Klinefelter syndrome: XXY karyotype obtained postmortem from cells from pericardial fluid. AB - A case of Klinefelter syndrome and a spontaneous cerebellar hemorrhage in a 12 year-old boy is presented. Autopsy revealed that the hemorrhage was due to the rupture of a dilated artery in an arteriovenous malformation in the right cerebellar hemisphere. The small, undescended testes exhibited partial atrophy of the seminiferous tubules. Postmortem chromosome analysis of cells from the pericardial fluid demonstrated a 47, XXY karyotype. He had previous surgical treatment for bilateral thumb polydactyly and patent ductus arteriosus. In juvenile cases of sudden death with overlapping morphological dysgenesis, postmortem karyotyping may provide important diagnostic information. PMID- 11005195 TI - Periadventitial extracranial vertebral artery hemorrhage in a case of shaken baby syndrome. AB - We report a case of a two-month-old boy who became unresponsive in the sole custody of his father. Resuscitation efforts on route to the hospital were able to restore the infant's heart beat. However, neurologic function never recovered. Autopsy revealed massive cerebral edema, recent subdural, and subarachnoid hemorrhages, bilateral retinal hemorrhages, and cervical spine ligament hemorrhages. Separation of individual cervical vertebrae showed extensive, bilateral, periadventitial vertebral artery hemorrhages between C1 and C4, with corresponding luminal compression of the vertebral arteries. The importance of this previously unreported phenomena of periadventitial vertebral artery hemorrhage in the setting of shaken baby syndrome is discussed. PMID- 11005196 TI - A non-fatal case of intoxication with foxglove, documented by means of liquid chromatography-electrospray-mass spectrometry. AB - The non-fatal self-poisoning of a 36-year-old female patient, who ingested a concoction of foxglove (Digitalis Purpurea), is presented. On the admission, initial symptoms were nausea and vomiting, abdominal pain, and cardiovascular shock with sinus bradycardia. Blood and urine were assayed for 17 cardiotonic hetorosides, using a highly specific LC-MS procedure. Serum and urine specimens were collected over five days and analyzed by liquid chromatography-electrospray mass spectrometry (LC-ES-MS). This accurate procedure allowed the determination of the digitalis glycosides and their metabolites in serum and urine. The serum concentrations of digitalis glycosides were maximum on the first day (gitoxin 13.1 ng/mL, digitoxin 112.6 ng/mL, digitoxigenin 3.3 ng/mL, and digitoxigenin mono-digitoxoside 8.9 ng/mL) and decreased over five days. We observed a peak gitaloxin level (112.6 ng/mL) on the fifth day only. After administration of atropine as well as dimeticone, alginic acid, and metoclopramide, health status improved. The peak urine concentrations were reached at hour 30 and were respectively 91.3 and 69.9 ng/mL for gitaloxin and digitoxin, while those of digitoxigenin, digitoxigenin mono-digoxoside and gitoxin were lower (respectively 0.7, 1, and 5.6 ng/mL). The patient was discharged on the fifth day when there were no residual symptoms. PMID- 11005197 TI - Distribution of amplified fragment length polymorphism D1S80 alleles in a Saudi Arabian population. PMID- 11005198 TI - Gene frequencies for four hypervariable DNA loci in a Chilean population of mixed ancestry. PMID- 11005199 TI - Mitochondrial DNA HVI and HVII variation in a north-east Spanish population. PMID- 11005200 TI - Commentary on Kayumov L, Pandi Perumal SR, Federoff P, Shapiro CM. Diagnostic values of polysomnograph in forensic medicine. J Forensic Sci 2000 Jan;45(1):191 4. PMID- 11005201 TI - Phenotypic differences at the HUMvWA locus amplified with different STR kits. PMID- 11005202 TI - Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. AB - Sucrose (Suc) plays a central role in plant growth and development. It is a major end product of photosynthesis and functions as a primary transport sugar and in some cases as a direct or indirect regulator of gene expression. Research during the last 2 decades has identified the pathways involved and which enzymes contribute to the control of flux. Availability of metabolites for Suc synthesis and 'demand' for products of sucrose degradation are important factors, but this review specifically focuses on the biosynthetic enzyme sucrose-phosphate synthase (SPS), and the degradative enzymes, sucrose synthase (SuSy), and the invertases. Recent progress has included the cloning of genes encoding these enzymes and the elucidation of posttranslational regulatory mechanisms. Protein phosphorylation is emerging as an important mechanism controlling SPS activity in response to various environmental and endogenous signals. In terms of Suc degradation, invertase-catalyzed hydrolysis generally has been associated with cell expansion, whereas SuSy-catalyzed metabolism has been linked with biosynthetic processes (e.g., cell wall or storage products). Recent results indicate that SuSy may be localized in multiple cellular compartments: (1) as a soluble enzyme in the cytosol (as traditionally assumed); (2) associated with the plasma membrane; and (3) associated with the actin cytoskeleton. Phosphorylation of SuSy has been shown to occur and may be one of the factors controlling localization of the enzyme. The purpose of this review is to summarize some of the recent developments relating to regulation of activity and localization of key enzymes involved in sucrose metabolism in plants. PMID- 11005203 TI - Ascorbic acid in plants: biosynthesis and function. AB - Ascorbic acid (vitamin C) is an abundant component of plants. It reaches a concentration of over 20 mM in chloroplasts and occurs in all cell compartments, including the cell wall. It has proposed functions in photosynthesis as an enzyme cofactor (including synthesis of ethylene, gibberellins and anthocyanins) and in control of cell growth. A biosynthetic pathway via GDP-mannose, GDP-L-galactose, L-galactose, and L-galactono-1,4-lactone has been proposed only recently and is supported by molecular genetic evidence from the ascorbate-deficient vtc 1 mutant of Arabidopsis thaliana. Other pathways via uronic acids could provide minor sources of ascorbate. Ascorbate, at least in some species, is a precursor of tartrate and oxalate. It has a major role in photosynthesis, acting in the Mehler peroxidase reaction with ascorbate peroxidase to regulate the redox state of photosynthetic electron carriers and as a cofactor for violaxanthin de-epoxidase, an enzyme involved in xanthophyll cycle-mediated photoprotection. The hypersensitivity of some of the vtc mutants to ozone and UV-B radiation, the rapid response of ascorbate peroxidase expression to (photo)-oxidative stress, and the properties of transgenic plants with altered ascorbate peroxidase activity all support an important antioxidative role for ascorbate. In relation to cell growth, ascorbate is a cofactor for prolyl hydroxylase that posttranslationally hydroxylates proline residues in cell wall hydroxyproline rich glycoproteins required for cell division and expansion. Additionally, high ascorbate oxidase activity in the cell wall is correlated with areas of rapid cell expansion. It remains to be determined if this is a causal relationship and, if so, what is the mechanism. Identification of the biosynthetic pathway now opens the way to manipulating ascorbate biosynthesis in plants, and, along with the vtc mutants, this should contribute to a deeper understanding of the proposed functions of this multifaceted molecule. PMID- 11005204 TI - Distribution of Tsc1 protein detected by immunohistochemistry in various normal rat tissues and the renal carcinomas of Eker rat: detection of limited colocalization with Tsc1 and Tsc2 gene products in vivo. AB - We and others previously demonstrated that hereditary mutation and a subsequent second hit in the rat homolog of tuberous sclerosis gene (Tsc2) are responsible for Eker renal carcinomas (RC). In humans, alteration in the TSC2 gene is known to cause the tuberous sclerosis complex (TSC) that results in hamartomatous lesions in multiple organs, but the function of TSC2 is not fully understood. In recent years, a second gene (TSC1) responsible for human TSC has been cloned, and binding between TSC1 and TSC2 proteins was reported. In this study, to clarify associations between Tsc proteins in vivo, the expression of Tsc1 protein was detected by immunohistochemistry, and compared with Tsc2 expression. Tsc1 protein was expressed in the nervous system and in many endocrine tissues, including pancreatic islets, the parathyroids, testis, and ovary. Tsc1 was also detected in the many epithelial tissues of organs, such as kidney, uterus, small and large intestine, and liver. Our results indicate overlapping, but not identical, organ distributions of Tsc1 and Tsc2 proteins. At the intracellular distribution, double fluorescent immunolabeling allowed the determination that only a partial portion of Tsc1 signals overlapped with Tsc2 in some organs. These results suggest the existence of co-localizing and independent forms of Tsc proteins in endogenous expressions. Additionally, relatively high expression of Tsc1 protein was detected in RC in the Tsc2 mutant (Eker) rat. PMID- 11005205 TI - Establishment of a novel endothelial target mouse model of a typhus group rickettsiosis: evidence for critical roles for gamma interferon and CD8 T lymphocytes. AB - A mouse model of typhus rickettsiosis that reproduces the hematogenous dissemination to the critical target organs, including brain, lungs, heart, and kidneys, primary endothelial and, to a lesser degree, macrophage intracellular rickettsial infection, and typical vascular-based lesions of louse-borne typhus and murine typhus was established. Intravenous inoculation of C3H/HeN mice with Rickettsia typhi caused disease with a duration of the incubation period and mortality rate that were dependent on the infective dose of rickettsiae. Lethal infection was associated with high concentrations of R. typhi in the lungs and brain, despite a brisker humoral immune response to the rickettsiae than in the sublethal infection. Gamma interferon and CD8 T lymphocytes were demonstrated to be crucial to clearance of the rickettsiae and recovery from infection in experiments in which specific monoclonal antibodies were administered to deplete these components. Death of animals depleted of gamma interferon or CD8 T lymphocytes was associated with overwhelming rickettsial infection demonstrated by titers of infectious rickettsiae and by immunohistochemistry. An effective antirickettsial immune response was associated with elevated serum concentrations of IL-12 on Day 5 and increased secretion of IL-12 by concanavalin-A-stimulated spleen cells on Day 5. Evidence for transient suppression of the immune response consisted of marked reduction in the secretion of IL-2 and IL-12 by concanavalin A-stimulated spleen cells on Days 10 and 15. This model offers excellent opportunities for study of attenuation and pathogenetic mechanisms of typhus rickettsiae, which are established biologic weapons of potential use in bioterrorism. PMID- 11005206 TI - Cytomegalovirus infection of the central nervous system stem cells from mouse embryo: a model for developmental brain disorders induced by cytomegalovirus. AB - Cytomegalovirus (CMV) is the most frequent infectious cause of developmental disorders of the central nervous system (CNS) in humans. Infection of the CNS stem cells seems to be primarily responsible for the generation of the brain abnormalities. In this study, we evaluated the infectivity of murine CMV (MCMV) in epidermal growth factor (EGF)-responsive CNS stem cells prepared from fetal mouse brains, and studied the effect of infection on growth and differentiation of the stem cells. The CNS stem cells were permissive for MCMV infection, although MCMV replication was slower than in mouse embryonic fibroblasts. MCMV infection inhibited the growth and DNA replication of the stem cells. A clonogenic assay revealed that MCMV infection suppressed generation of colonies from single stem cells. When uninfected stem cells were induced to differentiate, a decrease in expression of the primitive neuroepidermal marker nestin was observed by immunocytochemistry and flow cytometry, whereas expression of neurofilament and glial fibrillary acidic protein (GFAP) were induced. In virus infected CNS stem cells, nestin expression was retained, whereas the expression of neurofilament was more severely inhibited than that of GFAP in these cells. Two-color flow cytometry showed that differentiated glial precursor cells were preferentially susceptible to MCMV infection. MCMV-infected and uninfected CNS stem cells were transplanted into the neonatal rat brains. The reduced number of infected stem cells were engulfed into the subventricular zone and expressed GFAP, but did not migrate further, in contrast to the uninfected stem cells. These results suggest that suppression of the growth of the CNS stem cells and inhibition of the neuronal differentiation by CMV infection may be primary causes of disorders of brain development in congenital CMV infection. PMID- 11005207 TI - Lethal Mycobacterium bovis Bacillus Calmette Guerin infection in nitric oxide synthase 2-deficient mice: cell-mediated immunity requires nitric oxide synthase 2. AB - The role of nitric oxide (NO) in Mycobacterium bovis Bacillus Calmette Guerin (BCG) infection was investigated using nitric oxide synthase 2 (nos2)-deficient mice, because NO plays a pivotal protective role in M. tuberculosis infection. We demonstrate that nos2-deficient mice were unable to eliminate BCG and succumbed within 8 to 12 weeks to BCG infection (10(6) CFU) with cachexia and pneumonia, whereas all infected wild-type mice survived. The greatest mycobacterial loads were observed in lung and spleen. Nos2-deficient mice developed large granulomas consisting of macrophages and activated T cells and caseous necrotic lesions in spleen. The macrophages in granulomas from nos2-deficient mice had reduced acid phosphatase activities, suggesting that NO is required for macrophage activation. The absence of NOS2 affected the cytokine production of the Th1 type of immune response, except IL-18. Serum amounts of IL-12p40 were increased and IFN-gamma was decreased compared with wild-type mice. The lack of NOS2 resulted in an overproduction of TNF, observed throughout the infection period. Additionally, TNFR1 and TNFR2 shedding was altered compared with wild-type mice. Up-regulation of TNF may be compensatory for the lack of NOS2. The late neutralization of TNF by soluble TNF receptors resulted in heightened disease severity and accelerated death in nos2-deficient mice but had no effect in wild-type mice. In conclusion, the inability of nos2-deficient mice to kill M. bovis BCG resulted in an accumulation of mycobacteria with a dramatic activation of the immune system and overproduction of pro-inflammatory cytokines, which resulted in death. PMID- 11005208 TI - Analysis of the IGF axis in preneoplastic hepatic foci and hepatocellular neoplasms developing after low-number pancreatic islet transplantation into the livers of streptozotocin diabetic rats. AB - Preneoplastic hepatic foci have been demonstrated in liver acini, which drain the blood from intraportally transplanted pancreatic islets in streptozotocin-induced diabetic rats with mild persisting diabetes. In long-term studies of this animal model, hepatocellular adenomas and carcinomas (HCC) developed after a sequence of characteristic preneoplastic hepatic foci. In this experimental model, the local hyperinsulinism is thought to have a causative role. Because insulin and the insulin-like growth factor (IGF) axis are closely linked, an altered gene expression of the IGF axis components is likely. Therefore, preneoplastic hepatic foci and HCC were studied for the expression of IGF axis components. Glycogen storing "early" preneoplastic hepatic foci were detectable several days after pancreatic islet transplantation. Northern blot analysis, in-situ hybridization, and immunohistochemical studies of these "early" lesions demonstrated increased expressions of IGF-I and IGF binding protein-4 (IGFBP-4) in altered parenchymal cells, and a decreased expression of IGFBP-1. IGF-II was not detected in these preneoplastic foci. HCC arising in this model had decreased expressions of IGF-I and IGFBP-4 but IGFBP-1 expression was not significantly altered. Some HCC showed a more than 100-fold overexpression of IGF-II, whereas other tumors were completely negative for IGF-II expression. Low IGF-I receptor expression was detected in preneoplastic foci and adjacent nonaltered liver tissue. However, HCC tissue consistently showed an increased IGF-I receptor expression, rendering these tissues susceptible to the mitogenic effects of IGF. The altered gene expression in glycogen-storing preneoplastic hepatic foci, especially the up regulation of IGF-I and IGFBP-4 with the down-regulation of IGFBP-1, resemble the insulin-dependent regulation of these components in normal rat hepatocytes. These data agree with previous studies demonstrating a correspondence of the focal character, morphology, and enzyme pattern of preneoplastic hepatic foci with insulin effects on hepatocytes. The development from preneoplastic foci to HCC may be driven by insulin itself and/or an altered IGF axis component or yet unidentified factors. PMID- 11005209 TI - Suppression of Fas expression and down-regulation of Fas ligand in highly aggressive human thyroid carcinoma. AB - The Fas-FasL system seems to mediate thyrocyte death in Hashimoto's thyroiditis. In thyroid cancer, down-regulation of bcl-2 seems to alter apoptosis control. We compared the expression of immunoreactive Fas and FasL in normal thyroid with that of tumors ranging from benign to highly aggressive. Fas is essentially not expressed in normal thyrocytes, whereas FasL is expressed in approximately one third of cases. Expression of both markers is significantly up-regulated in adenoma and in well-differentiated papillary and follicular carcinoma. In contrast, Fas is suppressed and FasL is strongly reduced in the most aggressive histological variants (poorly differentiated and undifferentiated carcinoma). Immunohistochemistry findings have been confirmed by analysis of Fas-FasL mRNA transcripts. In vitro studies showed that the Fas receptor of thyroid tumor cells was functional, because apoptosis was induced by an agonistic Fas antibody. Fas expressing and Fas-resistant mammary cell lines were used as specificity controls. Together with our previous data inversely relating bcl-2 expression and thyroid tumor grade, the present findings further indicate that apoptotic pathways are altered in thyroid neoplasia. Thus, the Fas-FasL system may represent a marker of tumor aggressiveness. PMID- 11005210 TI - Expression and function of X chromosome-linked inhibitor of apoptosis protein in Sjogren's syndrome. AB - Apoptotic cell death in acinar and ductal epithelial cells is thought to play an important role in the development of salivary gland dysfunction in patients with Sjogren's syndrome (SS). We examined the expression of anti-apoptotic molecules in salivary glands from patients with SS. The labial salivary glands from six human T-cell leukemia virus (HTLV)-I-seronegative and eleven HTLV-I-seropositive SS patients were analyzed by immunohistochemistry. In vitro experiments were performed with a human salivary gland cell line (HSG cells). Immunohistologic analyses revealed that Bcl-2 and Bcl-x were preferentially expressed in salivary infiltrating mononuclear cells more than acinar and ductal epithelial cells. In contrast, strong X chromosome-linked inhibitor of apoptosis protein (XIAP) expression was evident in both acinar and ductal epithelial cells. The pattern of expression of these anti-apoptotic molecules was similar in both HTLV-I seropositive and HTLV-I -seronegative SS patients. Western blot analysis confirmed expression of XIAP in cultured HSG cells. The expression of XIAP in HSG cells was increased by IL-1beta, TGF-beta1, or IL-10. However, XIAP expression was down-regulated by TNF-alpha, which induced apoptotic cell death of HSG cells with an increase in caspase-3 activity. These effects of TNF-alpha in HSG cells were antagonized by IL-1beta, TGF-beta1, or IL-10. Our results suggest that XIAP is important in regulating apoptotic cell death of acinar and ductal epithelial cells in patients with SS. PMID- 11005211 TI - An immunocytochemical and in situ hybridization analysis of annexin 1 expression in rat mast cells: modulation by inflammation and dexamethasone. AB - The presence and localization of the anti-inflammatory protein annexin 1 (also known as lipocortin 1) in perivenular rat mast cells was investigated here. Using the rat mesenteric microvascular bed and a combination of morphologic techniques ranging from immunofluorescence to electron microscopy analyses, we detected the presence of annexin 1 in discrete intracellular sites, both in the nucleus and in the cytoplasm. In resting mast cells, most of the protein pool (approximately 80% of the cytosolic portion) was localized to cytoplasmic granules. In agreement with other cell types, treatment of rats with dexamethasone (0.2 mg/kg, ip) increased annexin 1 expression in mast cells, inducing a remarkable appearance of clusters of protein immunoreactivity. This effect was most likely the result of de novo protein synthesis as determined by an increase in mRNA seen by in situ hybridization. Triggering an ongoing experimental inflammatory response (0.3 mg of carrageenin, ip) increased annexin 1 mRNA and protein levels. In conclusion, we report for the first time the localization of annexin 1 in connective tissue mast cells, and its susceptibility not only to glucocorticoid hormone treatment, but also to an experimental acute inflammatory response. PMID- 11005213 TI - FISH analysis of gene aberrations (MYC, CCND1, ERBB2, RB, and AR) in advanced prostatic carcinomas before and after androgen deprivation therapy. AB - Genetic mechanisms leading to androgen-independent growth in advanced prostatic carcinomas (PC) are still poorly understood. Analysis of genes potentially involved in the regulation of tumor cell proliferation and apoptosis might confer better insight into this process and might lead to improved therapeutic strategies. Fluorescence in situ hybridization (FISH) analysis of dissociated nuclei with DNA probes for MYC (8q24)/#8, cyclin D1 gene (CCND1; 11q13)/#11, ERBB2 (17q13)/#17, the androgen receptor gene (AR; Xq12)/#X, and the retinoblastoma gene (RB; 13q14) was applied to formalin-fixed tissue from 63 patients with advanced PC after androgen deprivation therapy (ADT); matched tumor tissue before ADT was also available in 22 of these cases. The cut-points used were: "increased copy number," > or = 30% of all nuclei with increased FISH signals (centromere and/or gene); "amplification," > or = 15% of nuclei with "increased gene copy number." CCND1 and MYC gene "amplifications" were present before ADT in 25% and 33% of the cases, respectively; the frequency of these "amplifications" increased to 37% and 57% after ADT. Loss of the RB gene was nearly four times more frequent after ADT than before therapy (22% versus 6%). AR and ERBB2 gene "amplifications" occurred only after ADT in 36% and 30% of cases, respectively. With the exception of the AR gene, the copy number increase was low. After treatment, MYC and AR gene "amplifications" correlated with the proliferation rate (Ki-67/MIB1 index; p = 0.01 and p = 0.04), whereas ERBB2 "amplifications" were associated with increased apoptotic index (PCD/TUNEL; p = 0.016). However, no correlation between FISH results and clinical follow-up could be established. FISH analysis of genes putatively involved in PC progression revealed characteristic patterns of aberrations in advanced PC before and after ADT. Distinct changes in gene copy number before and after therapy suggests possible involvement of these genes in the escape from androgen control. PMID- 11005212 TI - The tyrosine kinase inhibitor tyrphostin AG 126 reduced the development of colitis in the rat. AB - Inflammatory bowel disease is characterized by oxidative and nitrosative stress, leukocyte infiltration, up-regulation of the expression of intercellular adhesion molecule-1 (ICAM-1), and up-regulation of P-selectin in the colon. Here we investigate the effects of the tyrosine kinase inhibitor, Tyrphostin AG 126, in rats subjected to experimental colitis. Colitis was induced in rats by intracolonic instillation of dinitrobenzene sulfonic acid (DNBS). Rats experienced hemorrhagic diarrhea and weight loss. Four days after administration of DNBS, the mucosa of the colon exhibited large areas of necrosis. Neutrophil infiltration (determined by histology as well as an increase in myeloperoxidase activity in the mucosa) was associated with up-regulation of ICAM-1 and P selectin, as well as high tissue levels of malondialdehyde. Immunohistochemistry for nitrotyrosine and poly(ADP-ribose) polymerase showed an intense staining in the inflamed colon. Staining with an anti-COX-2 antibody of sections of colon obtained from DNBS-treated rats showed a diffuse staining of the inflamed tissue. Furthermore, expression of inducible nitric oxide synthase was found mainly in macrophages located within the inflamed colon of DNBS-treated rats. Tyrphostin AG 126 (5 mg/kg daily ip) significantly reduced the degree of hemorrhagic diarrhea and weight loss caused by administration of DNBS. Tyrphostin AG 126 also caused a substantial reduction of (1) the phosphorylation of tyrosine residues of proteins (immunoblots of inflamed colon), (2) the degree of colonic injury, (3) the rise in myeloperoxidase activity (mucosa), (4) the increase in the tissue levels of malondialdehyde, (5) the increase in staining (immunohistochemistry) for nitrotyrosine and poly(ADP-ribose) polymerase, as well as (6) the up-regulation of ICAM-1 and P-selectin caused by DNBS in the colon. Thus, we provide the first evidence that the tyrosine kinase inhibitor Tyrphostin AG126 reduces the degree of colitis caused by DNBS. PMID- 11005214 TI - Expression of structurally unaltered androgen receptor in extramammary Paget's disease. AB - Extramammary Paget's disease (EMPD) is an uncommon neoplasm of the skin that shows differentiation to an apocrine sweat gland. Although we previously showed that erbB-2 overexpression may play a part in the progression of EMPD, molecular genetic defects underlying the development of EMPD are poorly understood. In the study described here, we examined androgen receptor expression and gene alterations in 30 cases of EMPD without internal malignancy. Immunohistochemistry revealed that 24 of 30 (80%) cases of EMPD variably expressed nuclear androgen receptor. Semi-quantitation of receptor content by scoring immunostained sections showed no difference between in situ (n = 17) and invasive (n = 13) EMPD tumors. Androgen receptor expression was also observed in four of six lymph node metastases. In these lymph nodes, expression of androgen receptor mRNA was confirmed by reverse transcriptase-polymerase chain reaction. Direct sequencing of exon 2 through exon 8, which encodes DNA- and hormone-binding domains of the androgen receptor gene, revealed no mutation in any of the 10 advanced stage tumors. Neither amplification nor deletion of the androgen receptor gene locus was detected by dual color fluorescence in situ hybridization analysis in 14 tumors. The present findings showing frequent expression of structurally unaltered androgen receptor in an advanced stage of EMPD may provide a rational basis for hormone therapy, which is widely used in the treatment of metastatic prostate cancer and androgen receptor-positive breast cancer recurrence. PMID- 11005215 TI - A novel method for the establishment of a pure population of nontransformed human intestinal primary epithelial cell (HIPEC) lines in long term culture. AB - A novel method for generating nontransformed human intestinal primary epithelial cell (HIPEC) lines in an in vitro culture system is reported here. Although several groups have reported the development of nontransformed intestinal epithelial cells (IEC) lines (Deveney et al, 1996; Latella et al, 1996; Pang et al, 1996; Perreault and Beaulieu, 1998), it still had been difficult to find an optimal condition to generate a pure population of nontransformed IEC in long term cultures. It was hypothesized that an appropriate growth factor/cytokine milieu that would mimic the physiological microenvironment might favor the survival of the isolated cells and might play a critical role in epithelial cell growth. To test this hypothesis, isolated progenitor/crypt cells were cultured in collagen-coated petri dishes in the presence of mucosal tissue-derived growth factor containing culture supernatants (14-18 hours) and a combination of hormonal supplements. Cell attachment and growth was observed within 24 hours and confluent monolayers were seen between 7 and 12 days. Immunofluorescence staining and flow cytometric analysis of the cells demonstrated positive staining with anti-cytokeratin-18 antibody confirming their epithelial origin. The reproducibility of the method has been confirmed by establishing a number of HIPEC lines from various segments of the gastrointestinal tract. This novel method of HIPEC line generation, which maximizes the similarity of the ex vivo culture system to in vivo conditions, will serve as a valuable tool for the establishment of a large number of HIPEC lines (intestinal epithelial cell bank) and for subsequent use in studies of the immunological/physiological epithelial function in the intestine. PMID- 11005216 TI - RT-PCR from archival single cells is a suitable method to analyze specific gene expression. PMID- 11005217 TI - Sociocultural perspectives on physical activity in the lives of older African American and American Indian women: a cross cultural activity participation study. AB - Illuminating the diversity and sociocultural specificity of women's experiences may be important if healthy lifestyles and quality of life are to be achieved. The incidence of cardiovascular disease linked to physical inactivity is high among African American and American Indian women. If more is understood about the experience of physical activity involvement, healthier living might be encouraged. Therefore, the purpose of this study was to explore the sociocultural meanings of physical activity for older (over the age of 40 years) African American and American Indian women who participated in the Cross Cultural Activity Participation Study (CAPS). Through qualitative in-depth interviews, we explored how sociocultural perspectives are related to perceptions about physical activity. Gender and other sociocultural factors influenced physical involvement on a continuum from negligible to significant. Both groups interviewed showed evidence that opportunities for physical activity in their free time did not always exist for them. For African American women, history and daily living issues were important factors limiting their involvement. Marginality limited American Indian women, but their cultural pride was often a source of physical activity. The juxtaposition of cultural and personal values emerged as a determinant of physical activity involvement among the women in this study. A further expansion of cultural and personal life situation perspectives is recommended to help understand the complex dimensions of physical activity as it relates to healthy living. PMID- 11005218 TI - Assessment of intimate partner violence among high-risk postpartum mothers: concordance of clinical measures. AB - Intimate partner violence (IPV) may be a more common problem than some conditions for which pregnant women are routinely screened and evaluated. Screening for IPV necessitates competence in assessment using reliable and valid instruments. The present study examines the concordance in the prevalence of abuse reported by 436 high-risk postpartum mothers using (1) a single item hospital screen, (2) trained paraprofessional interviewer-interview format, and (3) the Conflict Tactics Scale (CTS), a standardized research instrument. The hospital screen identified 4% of the sample as scared, threatened or hurt, the Family Stress Checklist (FSC) identified 16.9% of the fathers as having severe potential for violence, and the CTS identified 18.6% of the sample as victims of physical abuse at the hands of the baby's father. There was minimal overlap between the ratings of severe potential for violence and the occurrence of violence as assessed by the CTS. Study findings illustrate the inadequacy of current procedures for detecting IPV with high-risk populations and highlight the need to develop reliable and valid assessments that can be easily utilized in the hospital setting. PMID- 11005219 TI - Mothers with HIV/AIDS and their children: disclosure and guardianship issues. AB - For many mothers living with HIV/AIDS, whether, when, and how to disclose their HIV diagnosis to their children and arranging for future care are important although agonizing issues. Due to the increasing number of children who lose their mothers to AIDS and the dearth of empirical information about them, these issues are increasingly important to research. This study of 188 HIV-positive mothers and their 267 children of minor age in New York City revealed that only half the mothers had disclosed their HIV diagnosis to at least one of their children and only 57% had made formal plans for the children's care. As expected, older children were more likely to be informed than younger children. Contrary to some previous research, maternal disclosure was not related to ethnicity, advanced illness, improved psychological well-being, or greater or more satisfying social support resources. Implications for future research and provision of services to this group of women are discussed. PMID- 11005220 TI - Differences in fertility patterns between urban and rural women in Washington State, 1983-1984 to 1993-1994. AB - We use data on Washington State abortions and births for 1983-1984 and 1993-1994 to analyze trends for urban and rural women, using the demographic measures total abortion and total fertility rates. These express pacing of childbearing in a single number which is simple to calculate and interpret, and is age standardized. We find significant urban-rural differences. Total abortion rates decline and total fertility rates increase in both areas. However, the relative magnitudes of pacing decreases in abortions for rural women and increases in births for urban women are striking. The demographic measures are useful interpretive tools, and can be applied to a broad range of questions. PMID- 11005221 TI - The relationship of tamoxifen with dementia, depression, and dependence in activities of daily living in elderly nursing home residents. AB - OBJECTIVES: To evaluate the association between tamoxifen treatment and the prevalence of Alzheimer's disease, other dementias, depression, and the levels of dependence in activities of daily living (ADL) in elderly, nursing home residents. PARTICIPANTS: 93,031 women, at least 65 years old, whose data were part of the 1993 New York State MDS, and for whom there was documentation of treatment with at least one medication. SETTING: New York State long-term care facilities. DESIGN: Cross-sectional study via secondary analysis of 1,385 matched sets of residents. Each set included one resident who received tamoxifen treatment and up to four residents who did not. MEASUREMENTS: Included age, ethnicity, tamoxifen treatment, hormone replacement therapy, depression, levels of dependence in ADL and in cognitive skills for daily decision-making, and documentation of Alzheimer's disease and other dementias. RESULTS: Women receiving tamoxifen were less likely to have a diagnosis of Alzheimer's disease documented on the MDS (odds ratio: 0.674; 95% confidence interval: 0.551-0.823), were significantly more independent in bed mobility, eating, toileting (in each case P < 0.0001), personal hygiene (P = 0.0155), dressing (P = 0.0015), transferring (P = 0.0006), locomotion (P = 0.0016), and they had better cognitive skills for daily decision-making (P < 0.0001). They were, however, 42% more likely to have a MDS diagnosis of depression (P < 0.0001). All tests of statistical significance are two-sided. CONCLUSION: In this population, tamoxifen was associated with a higher level of independence in ADL and decision-making. While the MDS also suggested a relationship of tamoxifen with a lower prevalence of Alzheimer's disease, there seemed to be an increased prevalence of depression among treated women. The findings of this cross-sectional study could be evaluated in the randomized clinical trial that has just begun, and that will compare tamoxifen and raloxifene treatments. PMID- 11005222 TI - Exotic dancing and health. AB - The health and safety of women who work as exotic dancers are firmly embedded within the social organization of the strip club and the broader social, economic and political context of the work of exotic dancing. Exotic dancers in this study expressed health concerns associated with: the effects of costuming and appearance requirements; dirty work environments; problems due to stigmatization, sexual harassment and assault; and police disinterest or victim blaming. The balance between benefits and hazards related to exotic dancing is influenced not only by the personal choices made by dancers, but also by the organization of the strip club and the broader context within which exotic dancing takes place. PMID- 11005223 TI - Cell proliferation in the developing rat pineal gland. A bromodeoxyuridine immunohistochemical study. AB - The immunohistochemical detection of bromodeoxyuridine (BrdU) was used to study the cell proliferation in the developing rat pineal gland, from the appearance of pineal primordium in the embryonic day 15 (E15) until 30 days after birth. The results showed three different proliferative phases. From E15 to E21, the pineal gland shows a phase of rapid proliferation. The second phase corresponds to the first postnatal week, in which the number of labeled cells per surface unit decreases suddenly to values between 20% to 10% of those of embryonic period. From the second postnatal week onwards, the number of BrdU-positive cells progressively decreases. PMID- 11005224 TI - Glycogen autophagy in newborn rat hepatocytes. AB - Glycogen autophagy in newborn rat hepatocytes was studied by using enzyme determinations and electron microscopy. Cyclic AMP induced glycogen autophagy in these cells. Glycogen-hydrolyzing acid glucosidase activity increased whereas acid mannose 6-phosphatase activity decreased in the liver of these animals. Parenteral glucose, which prevents postnatal glucagon secretion and tissue cyclic AMP elevation, and propranolol which antagonizes cyclic AMP, inhibited glycogen autophagy. Glucosidase activity decreased and phosphatase activity increased. These findings raise the possibility that cyclic AMP-induced autophagic mechanisms in newborn rat hepatocytes are associated with changes in the activity of acid mannose 6-phosphatase. PMID- 11005225 TI - Calbindin D-28k immunoreactive nerve fibers in the carotid body of normoxic and chronically hypoxic rats. AB - The distribution and ultrastructural characteristics of calbindin D-28k immunoreactive nerve fibers were examined in the carotid body of the normoxic control rats by light and electron microscopy, and the abundance of calbindin D 28k fibers in the carotid body was compared in normoxic and chronically hypoxic rats (10% O2 and 3.0-4.0% CO2 for 3 months). Calbindin D-28k immunoreactivity was recognized in nerve fibers within the carotid body. Calbindin D-28k immunoreactive nerve fibers appeared as thin processes with many varicosities. They were distributed around clusters of glomus cells, and around blood vessels. Immunoelectron microscopy revealed that the calbindin D-28k immunoreactive nerve terminals are in close apposition with the glomus cells, and membrane specialization is visible in some terminals. Some dense-cored vesicles in the glomus cells were aggregated in this contact region. The chronically hypoxic carotid bodies were found to be enlarged several fold, and a relative abundance of calbindin D-28k fibers was lesser than in the normoxic carotid bodies. When expressed by the density of varicosities per unit area of the parenchyma, the density of calbindin D-28k fibers associated with the glomus cells in chronically hypoxic carotid bodies was decreased by 70%. These immunohistochemical findings indicate a morphological basis for involvement of calcium binding protein in the neural pathway that modulates carotid body chemoreception. PMID- 11005226 TI - Expression of a neu/c-erbB-2-like product in neuroendocrine cells of mammals. AB - The neu/c-erbB-2 oncogene encodes a 185 kDa protein closely homologous to the epidermal growth factor receptor. The protein product (p185) is a glycoprotein with an external domain and an internal domain with tyrosine kinase activity. Amplification and/or overexpression of p185 is related to several human adenocarcinomas. Subsequent studies demonstrated its presence in certain neuroendocrine (NE) neoplasms, including phaeochromocytomas, insulinomas and medullary thyroid carcinomas. However, relatively little is known about its role in normal cell growth regulation and development. Therefore, our objective was to determine whether neu/c-erbB-2 was expressed in normal NE tissues of different mammals, specially in humans, as it was in their neoplasms. We have examined by immunohistochemistry different endocrine glands (thyroid, pancreas, suprarrenal and hypophysis) and the small intestine of human beings, rats and guinea pigs, using two polyclonal antibodies raised against the intracytoplasmic part of the protein, and specific antigen absorption controls. We have found that a neu/c erbB-2-like product occurs in all normal NE tissues examined: C cells of the thyroid gland, chromaffin cells of the adrenal medulla, pancreatic islets, enteroendocrine cells of the small intestine and, finally, scattered cells of the adenohypophysis, according to a typical granular immunohistochemical pattern. Our results indicate that normal NE cells share a new common antigen in their cytoplasms, a neu/c-erbB-2-like product, with a similar immunostaining pattern to that presented by the neoplasms derived from them. PMID- 11005227 TI - Immunohistochemical detection of metallothionein in carcinomatous and normal human gastric mucosa. AB - Utilising a specific monoclonal mouse antibody (E9), metallothionein (MT) expression has been immunohistochemically investigated in 112 formalin-fixed paraffin-embedded surgical gastric samples, 38 of which were early carcinomas (EGC) and 74 advanced ones (AGC); clinico-pathological details and follow-up data (ranging from 3 to 197 months, mean 60.5 months) were available. Eighty-nine portions of gastric mucosa adjacent to examined carcinomas (transitional mucosa) were also analysed; in addition, 22 biopsies of normal gastric mucosa were studied as tissue control. The MT immunoreactivity was evaluated by staining and intensity-distribution scores. A various MT positivity was appreciable in the cytoplasm and nucleus of antrum or body gastric epithelial cells in 100% of normal control biopsies. 75/112 (67%) gastric carcinomas showed MT immunoreactivity with a significant lower expression in AGC. No relationships were encountered between MT immunostaining and clinico-pathological data; in addition, no difference in the Kaplan-Meier survival curves of patients with various MT expression was achieved. When the transitional mucosa was examined, 84/89 (94%) samples were stained although the immunoreaction was not always concordant with that encountered in adjacent carcinomatous elements. The significant statistical decrease of MT scores observed by us moving from normal to neoplastic gastric mucosa allows us to exclude the hypothesis of an overexpression of MT in gastric carcinomas. PMID- 11005228 TI - Changes of superoxide dismutase, catalase and glutathione peroxidase in the corneal epithelium after UVB rays. Histochemical and biochemical study. AB - In this study, the effects of UVA and UVB rays on antioxidant enzymes (superoxide dismutase, glutathione peroxidase, catalase) were examined in the corneal epithelium. The corneas of albino rabbits were irradiated with a UV lamp generating UVA (365 nm wavelength) or UVB rays (312 nm wavelength), 1 x daily for 5 min, from a distance of 0.03 m, over 4 days (shorter procedure) or 8 days (longer procedure). In contrast to UVA rays, which did not evoke significant disturbances, UVB rays changed the activities of antioxidant enzymes. The longer repeated irradiation with UVB rays was performed, the deeper the observed decrease in antioxidant enzymes. The shorter procedure evoked a more profound decrease of glutathione peroxidase and catalase (the enzymes cleaving hydrogen peroxide) than of superoxide dismutase, an enzyme scavenging superoxide radical and producing hydrogen peroxide during the dismutation reaction of a superoxide free radical. This may contribute to an insufficient hydrogen peroxide cleavage at the corneal surface and danger to the cornea from oxidative damage. After the longer procedure (UVB rays), the activities of all antioxidant enzymes were very low or completely absent. In conclusion, repeated irradiation of the cornea with UVB rays evokes a deficiency in antioxidant enzymes in the corneal epithelium, which very probably contributes to the damage of the cornea (and possibly also deeper parts of the eye) from UVB rays and the reactive oxygen products generated by them. PMID- 11005229 TI - Potential use of spectral image analysis for the quantitative evaluation of estrogen receptors in breast cancer. AB - Evaluation of estrogen receptor (ER) content is an important factor in the choice of therapy and prognosis of breast cancer patients. In this study, we demonstrate a new spectral image analysis technique for objective and quantitative evaluations of stained specimens. The SpectraCube system was used to analyze nuclear antigens in thirteen cases of breast cancer stained by the immunoperoxidase method with hematoxylin counterstain. Spectral imaging segregated the spectrum of diaminobenzidine (DAB) from the background color of hematoxylin and a spectral index was calculated. The spectral index essentially agreed with the pathologist's index (on a scale of 0 to 3) in seven out of the thirteen cases. A substantial number of ER positive pixels was detected in the two cases scored as 0 by the pathologist's index. In a test case scored as 1 by the pathologist's index we detected a significant number of pixels, representing 47% of the nuclei, with DAB-intensity values higher than the cut-off value of 1.2. These data suggest that spectral image analysis is a sensitive method providing intensive information with high reproducibility. Our spectral imaging method is highly flexible, enabling the user to define the spatial resolution of the analyzed specimen by choosing the number of pixels per one nucleus. PMID- 11005230 TI - Topographical difference of cytoskeletal organization in smooth muscle cells of rat duodenum revealed by quick-freezing and deep-etching method. AB - The sarcolemmal domain of rat duodenal smooth muscle cells includes caveolae and associated cytoskeletal or filamentous elements. We have used the quick-freezing, deep-etching method to examine the three dimensional relationships between these components. Replica membranes for separated strips of rat duodenal muscle layers were routinely prepared after extraction soluble proteins from cytoplasm and extracellular matrix. As results, 1) cytoskeletal elements in smooth muscle cells consisted mainly of striated thin filaments; 2) thin filaments were connected with some plasma membranes through filaments associated with the sarcolemma, which formed fine network structures beneath the sarcolemma; 3) many bridging structures between the filaments associated with the sarcolemma and the extracellular matrix were frequently detected in the plasma membrane; and 4) compact filaments associated with the sarcolemma almost disappeared near the caveolae, and only thin filaments were anchored to their neck parts. The special arrangement of the cytoskeletal components, which is probably necessary for the intestinal motility, characterizes the topographical difference of the smooth muscle sarcolemma. PMID- 11005231 TI - Suppressive effect of rebamipide, an antiulcer agent, against activation of human neutrophils exposed to formyl-methionyl-leucyl-phenylalanine. AB - Rebamipide, an antiulcer agent, has been shown to be able to prevent gastric mucosal injury resulting in part from activation of neutrophils. The mechanism of its suppressive action, however, remains to be established. The present study aimed to determine the effect of rebamipide on activation of isolated human neutrophils and to identify the signal transduction pathway involved in its regulation. In unstimulated cells, alkaline phosphatase activity was found residing in short rod-shaped intracellular granules. Upon stimulation with a chemotactic peptide formyl-methionyl-leucyl-phenylalanine, the granules fused to form elongated tubular structures and spherical vacuoles. Rebamipide inhibited reorganization of alkaline phosphatase-containing granules along with upregulation of alkaline phosphatase activity and CD16, a marker of the granules. It also suppressed chemotaxis, an increase in intracellular calcium ion concentration, and NADPH oxidase activation in cells stimulated with formyl methionyl-leucyl-phenylalanine. In contrast, the drug showed no inhibitory action toward upregulation of alkaline phosphatase activity and CD16, and activation of NADPH oxidase in cells stimulated with phorbol myristate acetate, an activator of protein kinase C. These findings demonstrate that rebamipide exerts a broad spectrum of suppressive actions toward biological functions of human neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine, but not with phorbol myristate acetate, and suggest that the upstream point of protein kinase C is the signal transduction pathway involved in its regulation. PMID- 11005232 TI - Dendritic cells and interleukin-2: cytochemical and ultrastructural study. AB - The aim of the present study was to verify the effect of IL-2 on dendritic cell (DC) differentiation. Various cytokines have been indicated as factors inducing DC differentiation, but no data about the interleukin-2 (IL-2) effect on DC differentiation have been reported. Monocytes isolated from peripheral blood were treated in vitro with the following factors: IL-2, IL-4, GM-CSF and G-CSF alone or in combination. Morphological (also ultrastructural) and cytochemical observations were carried out starting from 3 to 21 days of treatment. The results indicate that the differentiation of cells showing dendritic pattern is related to the presence of IL-2. Moreover a synergic effect of IL-2 and GM-CSF was observed. The enzymatic features changed with the culture time: before the differentiation into DC, the stimulated cells expressed the typical pattern of monocytes. On the contrary, at advanced stage of differentiation, some enzyme activities changed and in terminally differentiated dendritic cells the reactions for peroxidase and serine esterase were negative. Considering the morphological features, the ability to interact with lymphocytes and the enzymatic pattern observed, we suppose that IL-2 may act as a maturative factor rather than as a growth factor in the DC differentiation. PMID- 11005233 TI - Age-related morphometric changes in the pineal gland. A comparative study between C57BL/6J and CBA mice. AB - Relatively little is known about the effects of melatonin on the aging of the pineal, the organ which is the main place for synthesis of this hormone. Using simple morphometric methods, some parameters of the pineal gland, such as total volume, number of pinealocytes and pinealocyte volume were estimated in two mice strains: normal CBA and melatonin-deficient C57BL/6J. Two age groups, 6 weeks and 10 months, were studied in order to evaluate possible differential age-related changes between both strains. Pineals of both strains have similar morphometric and morphological features at 6 weeks of age. This suggests that pineal development, which has already concluded at 6 weeks of age, is not affected by the absence of melatonin synthesis in the pinealocytes. Later on, CBA pineal showed an increase in size caused by cellular hypertrophy. In contrast, the C57BL/6J pineal volume decreased by loss of pinealocytes in the same period of time. Semithin sections analysed by light microscopy did not show that this cell death was evident in the C57BL/6J strain at any of the ages studied. Thus, a gradual loss of pinealocytes could be hypothesised in these pineals. These results suggest that pineal melatonin could have a role in the maintenance of pinealocyte viability and the increase of pineal size which takes place after development. The abnormal pattern observed in the C57BL/6J pineal should be taken into account in future studies on this gland. PMID- 11005234 TI - Intrinsic innervation in the intestine of the lizard Podarcis hispanica. AB - The aim of this study was the description of the morphology and distribution of nerve structure elements in the intestine of the lizard Podarcis hispanica using different histochemical methods; namely acetylcholinesterase (AChE), formol induced fluorescence for catecholamines (FIF), nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d), and immunohistochemistry for vasoactive intestinal peptide (VIP), as well as substance P (SP) and electron microscopy. The AChE method showed fibres in the myenteric and submucosal plexus, with a higher fibre density in the large intestine. The highest number of related neurons was located in the myenteric plexus ganglia. Noradrenergic innervation was distributed through the myenteric and submucosal plexus, and also around blood vessels, with the highest fibre density in the large intestine. VIP immunohistochemistry showed a wide distribution of positive fibres throughout the intestine, although the highest density was again detected in the large intestine. Small positive cells for VIP were located at internodal segments in the plexus. SP labeling, although subtle, was present all along the intestine. It showed delicate varicose nets and few fibres innervating blood vessels. Small positive cells for SP were located in the large intestine. The indirect method to detect nitric oxide (NO)-producing system showed neural cells in the myenteric plexus ganglia of the large intestine. Electron microscopy showed ganglion neurons with scattered chromatin condensations, glial cells with higher electron density, and axons with varicosities occupied by different vesicles. We also identified certain cells as interstitial cells of Cajal due to their ultrastructural features. They were mostly located in the region of the myenteric plexus. PMID- 11005235 TI - Nuclear and cytoplasmic lectin receptor sites in rat Py1a osteoblasts. AB - The intracellular distribution of lectin receptor sites was studied in the rat Pyla osteoblasts using immunofluorescence at the confocal microscopy level. This immortalized cell line was found to represent a satisfactory model to study the occurrence and distribution of sugar moieties. Our data showed distinct affinity patterns of lectins recognizing different terminal or internal sugar residues. For some lectins, the binding patterns appeared to be cell cycle-independent, whereas for PNA the cell cycle greatly influenced the nuclear binding. By combining lectin affinity with sialidase degradation and alcoholic saponification the sialic acid acceptor sugars and derivatives were also visualized. In particular, glycoconjugates with sialic acids linked to beta-galactose, and mainly C4 acetylated, were located in the cytoplasm, while glycoconjugates characterized by sialic acids linked to alpha-N-acetylgalactosamine, and devoid of acetyl groups at C4, were almost exclusively found in the nucleus. The comparison of lectin affinities, with and without prior glycosidase digestions, allowed us to gain further insight into the chemical composition of glycoconjugates that act as the lectin receptor sites that appeared to belong to O- and N-linked glycoconjugates. The use of additional enzymatic treatments were useful to better establish the localization of nuclear receptor sites and results were compared with previous studies about endogenous and exogenous lectins in an attempt to reconcile the association of lectins and sugars within the nucleus and their possible involvement in modulation of cell proliferation and/or response to chemical signals. The above digestions also provided information about the cytoplasmic binding patterns. PMID- 11005236 TI - Confocal evaluation of native and induced lectin binding contributes to discriminate between lingual gland glycocomponents in quail. AB - A confocal analysis was performed on the quail (Coturnix coturnix japonica) lingual salivary glands where the carbohydrate chains were studied by lectin histochemistry. For this purpose, appropriate FITC- and TRITC-conjugates were used for double binding also accomplished with sialidase digestion. The glycosidic components of the quail lingual salivary glands were found to be heterogeneously distributed on the different secretory structures as well as on the single secretory elements of each adenomere. The rostral portion of the anterior lingual gland was found to only secrete neutral glycocomponents, characterized by terminal beta-galactose, N-acetylgalactosamine and fucose residues in contrast to the caudal portion that was shown to be extremely heterogeneous and to produce sialylated glycoconjugates characterized by the terminal sequences sialic acid-beta-galactose-N-acetylgalactosamine, sialic acid beta-galactose-N-acetylglucosamine, and sialic acid-alpha-N-acetylgalactosamine partly codistributed within secretory adenomeres. The posterior lingual gland was observed to be the major contributor to the secretion of salivary mucins containing sialoglycoconjugates with terminal sialic acid residues linked to beta galactose-N-acetylgalactosamine or alpha-N-acetylgalactosamine often located in distinct secretory elements. PMID- 11005237 TI - Selective expression of lysyl oxidase (LOX) in the stromal reactions of broncho pulmonary carcinomas. AB - Lysyl oxidase (LOX) is the extracellular enzyme that initiates the main pathway of collagen and elastin cross-linking. LOX has also been correlated with the ras recision gene, a putative tumour suppressor isolated from revertants of ras transformed fibroblasts. The present study investigates the potential correlation of LOX-dependent matrix protein cross-linking in the stromal reaction of lung carcinomas, with reference to the architecture of the main stromal reactions accompanying the neoplastic breast tissues. A strong LOX expression was associated with the hypertrophic scar-like stromal reaction found at the front of tumour progression in squamous carcinomas, adenocarcinomas, large cell carcinomas, or at sites of initial extense in bronchiolo-alveolar carcinomas. In contrast, little or no LOX expression was found within the stromal reaction of invasive carcinomas, small cell carcinomas, and neuro-endocrine carcinomas. The significance of LOX expression and of the stromal reaction are discussed, in light of data that associate LOX expression with tumours displaying a rather good prognosis. PMID- 11005238 TI - Morpho-histochemical study of the biological effects of sodium dodecyl sulphate on the digestive gland of the Portuguese oyster. AB - In the present work we have studied the effects on survival as well as histopathological and histochemical alterations on the digestive gland of the Portuguese oyster (Crassostrea angulata, Lmk.) induced by acute action of the anionic tensioactive Sodium Dodecyl Sulphate (SDS) at different concentrations (50, 100, 200 and 400 mg/L). Firstly, the LC 50 at 96 h was found to be 136 mg/L SDS. Secondly we calculated the exposure time required for 50% mortality of the specimens exposed to different concentrations, and thirdly we examined histopathological alterations (degenerative processes ranging from inflammatory responses to extreme vacuolation) and histochemical changes in the distribution of carbohydrates and proteins that appeared at each employed concentration. The degree of these alterations and the effects on survival were dependent upon the SDS concentration. The above established modifications indicate that exposure to SDS has a detrimental effect on oyster digestive gland, perturbing metabolic and nutritional functions, what may have some influence on oyster survival. PMID- 11005239 TI - Melanization stimulating factors in the integument of the Mugil cephalus and Dicertranchus labrax. AB - The pigment pattern expression resides in the chromatoblasts of the embryonic skin. The differentiation of these chromatoblasts is influenced by specific local factors such a melanization inhibiting factor (MIF) and a melanization stimulating factor (MSF). We reveal the presence of these factors by means of a series of experiments on the skin of the marine species of fish Dicertranchus labrax and Mugil cephalus, each with different pigment pattern, the former having a light skin and the latter a darker one. Media conditioned by exposure to dorsal and/or ventral skin, stimulates the melanization of Xenopus laevis neural crest cells throughout a 3 day assay period. Similarly conditioned culture media tested on B16-F10 murine malignant melanocytes, revealed a considerable influence in enzymatic activities: dopachrome tautomerase (DCT), tyrosine hydroxylase and dopa oxidase. The use of media in a dose response basis suggests that the conditioned media may contain both melanophore stimulating and inhibiting factors. The results obtained may actually reflect the resultant activity of the two factors present. PMID- 11005240 TI - Molecular actions of nitric oxide in mesangial cells. AB - Nitric oxide (NO) is a widely recognized mediator of physiological and pathophysiological signal transmission. Its generation through L-arginine metabolism is relevant in the mesangium of the kidney where NO is produced by constitutive and inducible NO-synthase isoenzymes. Signaling is achieved through target interactions via redox and additive chemistry. In mesangial cells (MC), the outcome of these modifications promote on one side activation of soluble guanylyl cyclase while on the other side cytotoxicity is elicited. These contrasting situations are characterized by: 1) cGMP formation and signal propagation towards myosin light chain kinase, the effector system that regulates F-actin assembly, thereby affecting reversible relaxation/contraction of mesangial cells; and 2) initiation of morphological and biochemical alterations that are reminiscent of apoptosis such as chromatin condensation, p53 or Bax accumulation as well as caspase-3 activation. Off note, NO formation with concomitant initiation of apoptosis is efficiently antagonized by the simultaneous presence of superoxide (O2-). We will recall the consequences that stem from a diffusion controlled NO/O2- interaction thereby redirecting the apoptotic initiating activity of either NO or O2- towards protection. The crosstalk between cell destructive and protective signaling pathways, their activation or inhibition under the modulatory influence of NO will be discussed. Here we give examples of how NO elicits physiological and pathophysiological signal transmission in rat MC. PMID- 11005241 TI - Liver fibrosis, the hepatic stellate cell and tissue inhibitors of metalloproteinases. AB - Liver fibrosis occurs as a consequence of net accumulation of matrix proteins (especially collagen types I and III) in response to liver injury. The pathogenesis of liver fibrosis is underpinned by the activation of hepatic stellate cells (HSC) to a myofibroblast like phenotype with a consequent increase in their synthesis of matrix proteins such as interstitial collagens that characterise fibrosis. In addition to this there is increasing evidence that liver fibrosis is a dynamic pathologic process in which altered matrix degradation may also play a major role. Extracellular degradation of matrix proteins is regulated by matrix metalloproteinases (MMPS)- produced by HSC--which in turn are regulated by several mechanisms which include regulation at the level of the gene (transcription and proenzyme synthesis), cleavage of the proenzyme to an active form and specific inhibition of activated forms by tissue inhibitors of metalloproteinases (TIMPS). Insights gained into the molecular regulation of HSC activation will lead to therapeutic approaches in treatment of hepatic fibrosis in the future, and could lead to reduced morbidity and mortality in patients with chronic liver injury. PMID- 11005242 TI - Integrin activation by chemokines: relevance to inflammatory adhesion cascade during T cell migration. AB - The adhesive function of integrins is regulated through cytoplasmic signaling induced by several stimuli, whose process is designated "inside-out signaling". A large number of leukocytes are rapidly recruited to the sites of inflammation where they form an essential component of the response to infection, injury, autoimmune disorders, allergy, tumor invasion, atherosclerosis and so on. The recruitment of leukocytes into tissue is regulated by a sequence of interactions between the circulating leukocytes and the endothelial cells. Leukocyte integrins play a pivotal role in leukocyte adhesion to endothelial cells. During the process, the activation of integrins by various chemoattractants, especially chemokines, is essential for integrin-mediated adhesion in which a signal transduced to the leukocyte converts the functionally inactive integrin to an active adhesive configuration. We have proposed that H-Ras-sensitive activation of phosphoinositide 3 (PI 3)-kinase and subsequent profilin-mediated actin polymerization, can be involved in chemokine-induced integrin-dependent adhesion of T cells. The present review documents the relevance of cytoplasmic signaling and cytoskeletal assembly to integrin-mediated adhesion induced by chemoattractants including chemokines during inflammatory processes. In contrast, various adhesion molecules are known to transduce extracellular information into cytoplasm, which leads to T cell activation and cytokine production from the cells, designated "outside-in signaling". Such a bi-directional "cross-talking" among adhesion molecules and cytokines is most relevant to inflammatory processes by augmenting immune cell migration from circulation into inflamed tissue such as rheumatoid arthritis, tumor invasion, Behcet's disease and atherosclerosis. PMID- 11005243 TI - Molecular genetic approaches to microtubule-associated protein function. AB - Protein function in vivo can be studied by deleting (knock-out) the gene that encodes it, and search for the consequences. This procedure involves different technologies, including recombinant DNA procedures, cell biology methods and histological and immunocytochemical analysis. In this work we have reviewed these procedures when they have been applied to ascertain the function of several microtubule-associated proteins. These proteins have been previously involved, through in vitro experiments, in having a role in the microtubule stabilization. Here, we will summarize the generation and characterization of different microtubule-associated protein knock-out mice. Special attention will be paid to MAP1B knock-out mice. Amongst the different MAPs knock-out mice these show the strongest phenotype, the most likely for being MAP1B, the MAP that is expressed earliest in neurogenesis. Molecular genetics could be considered as a valid and useful procedure to truly establish the in vivo functions of a protein, although it is necessary to be aware of possible artifacts such as the generation of some kinds of RNA alternative splicing. To avoid this the best strategy to be used must consider the deletion of the exon that contains the functional domains of the protein. PMID- 11005244 TI - Clinical applications of image cytometry to human tumour analysis. AB - Image cytometry (ICM) is widely applied to the automated screening, the detection, the diagnosis, the classification, the prognosis and the therapeutic follow-up of different types of cancers (breast, bladder, cervix,...). This review describes the analysis methods and the applications of nuclear image analysis, the determination of DNA content and the analysis of morphometry and of nuclear texture. DNA content analysis can contribute to a prognostic information in addition to other prognostic factors for breast, renal and prostate cancers. For ovarian cancer, aneuploidy seems to be related to prognosis. Bladder tumours with DNA aneuploidy were frequently of high malignancy while ploidy was significantly correlated to relapse risk. For digestive cancers, patients presenting DNA diploid tumours show a better survival than patients with aneuploid ones. Morphometry seems to be a more important criterion than other conventional prognostic factors of invasive breast and digestive carcinomas. A differential diagnosis between normal and neoplastic thyroids is more precise when based on a quantitative evaluation of texture associated to morphometry. Textural parameters permit the discrimination of two populations of patients having a different prognosis and could thus be an aid for prognosis in prostatic cancers. Morphonuclear parameters contribute to separate low and high grade bladder carcinomas. Although ICM was frequently reported, results from the reported examples were not always obvious. In conclusion, the measurements obtained with ICM could be helpful for a decision in several cancers but could not be a substitute for the classical approach of the pathologist. PMID- 11005245 TI - Adoptive cellular immunotherapy: NK cells and bone marrow transplantation. AB - Allogeneic bone marrow transplantation (BMT) has been increasingly used for the treatment of both neoplastic and non-neoplastic disorders. However, serious obstacles currently limit the efficacy and thus more extensive use of BMT. These obstacles include: graft-versus-host disease (GVHD), relapse from the original tumor, and susceptibility of patients to opportunistic infections due to the immunosuppressive effects of the conditioning regimen. Overcoming these obstacles is complicated by dual outcome of existing regimens; attempts to reduce GVHD by depleting T cells from the graft, result in increased rates of tumor relapse and failure of engraftment. On the other hand, efforts to increase graft-versus-tumor (GVT) effects of the transplant also promote GVHD. In this review, the use of natural killer (NK) cells to overcome some of these obstacles of allogeneic BMT is evaluated. Adoptive immunotherapy using NK cells after allogeneic BMT has several potential advantages. First, NK cells can promote hematopoiesis and therefore engraftment by production of hematopoietic growth factors. Second, NK cells have been shown to prevent the incidence and severity of GVHD. This has been shown to be at least partially due to TGF-beta, an immunosuppressive cytokine. Third, NK cells have been shown to augment numerous anti-tumor effects in animals after BMT suggesting a vital role of NK cells in mediating GVT effects. Finally, NK cells have been demonstrated to affect B cell recovery and function in mice. Therefore, understanding the mechanisms of beneficial effects of NK cells after BMT may lead to significant increases in the efficacy of this procedure. PMID- 11005246 TI - Cell proliferation and apoptosis in prostate cancer: significance in disease progression and therapy. AB - Recent biochemical and genetic studies have substantially increased our understanding of death signal transduction pathways, making it clear however, that apoptosis is not a single-lane, one-way street. Rather, multiple parallel pathways have been identified. For instance, analysis of bcl-2, bax, p53, and caspase knockout mice while establishing distinct roles for each of these apoptotic players, they also provided valuable information for the design of specific inhibitors of apoptosis. Thus blocking one pathway, as in caspase knockout mice, what we observe is not a complete suppression of apoptosis but rather a delay in apoptosis induction (Hakem et al., 1998; Kuida et al., 1998). In view of nature's means of ensuring activation of a compensatory apoptotic response, when one pathway fails in developing prostate cancer therapeutic interventions, the challenge remains to further dissect individual apoptotic pathways. Advances in our understanding of the integrated functions governing prostate cell proliferation and cell death, clearly suggest that effective prostate cancer therapies are not only molecularly targeted, but that are also customized to take into account the delicate balance of opposing growth influences in the ageing gland. In this review we discuss the evidence on the significance of molecular deregulation of the key players of this growth equilibrium, apoptosis and cell proliferation in prostate cancer progression, and the clinical implications of changes in the apoptotic response in disease detection and therapy. PMID- 11005247 TI - Use of interphase cytogenetics in demonstrating specific chromosomal aberrations in solid tumors--new insights in the pathogenesis of malignant melanoma and head and neck squamous cell carcinoma. AB - The detection of structural and numerical chromosomal aberrations is an important part of the characterization of tumors and genetic diseases. The direct demonstration of DNA sequences in interphase nuclei and metaphases by fluorescence in situ hybridization (FISH) has been termed interphase cytogenetics. It has been proven as a powerful technique to detect specific aberrations in a wide variety of cell types, including paraffin-embedded tissue. Nowadays a standard method in leukemia and lymphoma, interphase cytogenetics contributes mainly to the diagnosis in these tumors and helps to classify soft tissue tumors. Therefore FISH is mandatory for the choice of therapy in these tumors. In contrast to the aforementioned, up to now, the value of FISH in solid tumors is mostly limited to pure research and contributes in this way to our understanding of tumor biology. But with the use of paraffin-embedded tissue and the first results obtained, it seems very likely that a direct correlation between histological classification and cytogenetic characteristics of solid tumors can be achieved in the near future. This information might not only provide insights into tumor biology, but could also contribute to a different tumor classification, a sort of risk estimation, where we might predict the possible biological behavior of solid tumors. This could greatly influence further therapeutic decisions thus establishing the FISH technique as an indisputable part in the diagnosis of solid tumors. PMID- 11005248 TI - Gene therapy strategies for intracranial tumours: glioma and pituitary adenomas. AB - Intracranial tumours such as brain gliomas and pituitary adenomas pose a challenging area of research for the development of gene therapy strategies, both from the point of view of the severity of the diseases, to the physiological implication of gene delivery into the central nervous system and pituitary gland. On the one hand, brain gliomas are very malignant tumours, with a life expectancy of six months to a year at the most after the time of diagnosis, in spite of advances in treatment modalities which involve chemotherapy, surgery and radiotherapy. Gene therapy for these tumours is therefore a very attractive therapeutic modality which due to the severity of the disease is already in clinical trials. On the other hand, pituitary tumours are usually benign, and in most cases, treatment is successful. Nevertheless, there are some instances, especially with the macroadenomas and some invasive tumours in which treatment fails. Gene therapy strategies for these adenomas therefore needs to progress substantially in terms of safety, adverse side effects and physiological impact on the normal pituitary gland before clinical implementation. In this paper, we will review gene delivery systems both viral and non-viral and several therapeutic strategies which could be implemented for the treatment of these diseases. These include cytotoxic approaches both conditional and direct, immune stimulatory strategies, anti-angiogenic strategies and approaches which harness pro-apoptotic and tumour suppressor gene targets. We will also review the models which are currently available in which these gene therapy strategies can be tested experimentally. This new therapeutic modality holds enormous promise, but we still need substantial improvements both from the delivery, efficacy and safety stand points before it can become a clinical reality. PMID- 11005249 TI - Gene therapy using herpes simplex virus-based vectors. AB - Gene therapy involves the use of specific genes to treat human diseases and is thus critically dependent on efficient gene delivery systems. Although a variety of systems for such gene delivery are under development, HSV has unique advantages in terms of its large genome size and for gene delivery in the nervous system because of its ability to enter a latent state in neuronal cells. Considerable progress has been made in the effective disablement of this virus whilst retaining its ability to deliver genes and in producing long term expression of the foreign gene. Although much remains to be achieved in the further disablement of the virus and its testing in rodent and primate models of human diseases, it is likely that these viruses may ultimately be of use in human gene therapy procedures particularly for otherwise intractable neurological diseases. PMID- 11005250 TI - Localization of androgen and estrogen receptors in rat and primate tissues. AB - There is now evidence that estrogens and androgens are exerting their effects in different tissues throughout the body. In order to determine the sites of action of these steroids, studies have been performed to identify at the cellular level the localization of androgen receptor (AR) and the two estrogen receptor (ER) subtypes, ERalpha and ERbeta, specially in the rat, monkey and human. In the prostate, AR was observed in the secretory and stromal cells. In the testis, Sertoli, Leydig and myoid cells were labelled. In the epididymis and seminal vesicles, both epithelial and stromal cells contained AR. In the ovary, AR was detected in granulosa and interstitial cells. In the uterus, epithelial, stromal and muscle cells were all immunopositive for AR. In the central nervous system, AR-containing neurons were found to be widely distributed throughout the brain. In the mammary gland, epithelial cells in acini and ducts and stromal cells were demonstrated to express AR. In the skin, AR was detected in keratinocytes, sebaceous and sweat glands, and hair follicles. In addition, AR was also found in anterior pituitary, thyroid, adrenal cortex, liver, kidney tubules, urinary bladder, cardiac and striated muscle, and bone. The ER subtypes are in general differentially expressed. While ERalpha has been predominantly found in anterior pituitary, uterus, vagina, testis, liver and kidney, ERbeta is predominant in thyroid, ovary, prostate, skin, bladder, lungs, gastro-intestinal tract, cartilage and bone. In tissues which contain both receptor subtypes, such as ovary, testis and various regions of the brain, a cell-specific localization for each ER subtype has been generally observed. Altogether, the recent results on the cellular localization of sex steroid receptors will certainly contribute to a better understanding of the specific role of these steroids in different target organs. PMID- 11005251 TI - Identification and characterization of genes responsive to apoptosis: application of DNA chip technology and mRNA differential display. AB - Apoptosis (programmed cell death) is a genetically programmed active cell death process for maintaining homeostasis under physiological conditions and for responding to various stimuli. Many human diseases have been associated with either increased apoptosis (such as AIDS and neurodegenerative disorders) or decreased apoptosis (such as cancer and autoimmune disorders). In an attempt to understand apoptosis signaling pathway and genes associated with apoptosis, we established two cell model systems on which apoptosis is induced either by DNA damaging agent, etoposide or by redox agent, 1,10-phenanthroline (OP). DNA chip profiling or mRNA differential display (DD) was utilized to identify genes responsive to apoptosis induced by these two agents. In etoposide model with chip hybridization, we defined signaling pathways that mediate apoptosis in p53 dependent manner (through activation of p53 target genes such as Waf-1/p21, PCNA, GPX, S100A2 and PTGF-beta) as well as in p53-independent manner (through activation of ODC and TGF-beta receptor, among others). In OP model with DD screening, we cloned and characterized two genes: glutathione synthetase, encoding an enzyme involved in glutathione synthesis and Sensitive to Apoptosis Gene (SAG), a novel evolutionarily conserved gene encoding a zinc RING finger protein. Both genes appear to protect cells from apoptosis induced by redox agents. Further characterization of SAG revealed that it is a growth essential gene in yeast and belongs to a newly identified gene family that promotes protein ubiquitination and degradation. Through this activity, SAG regulates cell cycle progression and many other key biological processes. Thus, SAG could be a valid drug target for anti-cancer and anti-inflammation therapies. PMID- 11005252 TI - Considerations on the thalamostriatal system with some functional implications. AB - The thalamostriatal projections are largely neglected in current reviews dealing with basal ganglia function. In the past few years, however, several studies have re-evaluated these projections and have postulated their implication in more complex tasks within the basal ganglia organization. In this review, we try to focus on the morphological and functional importance of this system in the basal ganglia of the rat, cat and monkey. Special attention is paid to the thalamus as an important place for interaction between the input and the output systems of the basal ganglia through the thalamostriatal projections. Thus, we stress on the overlapping thalamic territories between the thalamic projection of the output nuclei of the basal ganglia and the thalamostriatal neurons. Our experimental data support the existence of several thalamic feedback circuits within the basal ganglia functional design. Finally, some considerations are provided upon the functional significance of these thalamic feedback circuits in the overall organization of the basal ganglia in health and disease. PMID- 11005253 TI - Biological and clinical review of stromal tumors in the gastrointestinal tract. AB - Submucosal tumors of the gastrointestinal tract (GI tract) mainly consist of gastrointestinal mesenchymal tumors (GIMTs) that are distributed in the GI tract from the esophagus through the rectum. GIMTs include myogenic tumors, neurogenic tumors and gastrointestinal stromal tumors (GISTs). The term "GIST" is now preferentially used for the tumors that express CD34 and KIT. GIMTs are composed of spindle or epithelioid cells, and 20% to 30% show malignant behavior, including peritoneal dissemination and hematogenous metastasis. KIT expression and mutations in the c-kit gene are found only in GISTs, but not in myogenic or neurogenic tumors. Mutation in the c-kit gene is associated with aggressive features and poor prognosis, and malignant GISTs frequently have mutations in the c-kit gene. The clinicopathological features of GISTs with or without c-kit mutations are markedly different. Therefore, GIMTs may be divided into four major categories based on histochemical and genetic data: myogenic tumors; neurogenic tumors; GISTs with c-kit mutation; and GISTs without c-kit mutation. The origin of GISTs is not fully understood. However, phenotypical resemblance to the interstitial cells of Cajal (ICCs) and gain-of-function mutations in the c-kit gene may suggest origin from ICCs and/or multipotential mesenchymal cells that differentiate into ICCs. PMID- 11005254 TI - Signaling pathways mediated by tumor necrosis factor alpha. AB - Tumor necrosis factor alpha (TNFalpha) has been shown to trigger many signaling pathways. Following oligomerization by TNFalpha, the receptors TNF-RI and TNF-RII associate with adapter molecules via specific protein-protein interactions. The subsequent recruitment of downstream molecules to the receptor complex enables propagation of the TNFalpha signal. Two cellular responses to TNFalpha have been well documented, the induction of cell death and the activation of gene transcription for cell survival. TNFalpha-induced apoptosis involves the activation of caspase cascades, which culminate in the cleavage of specific cellular substrates to effect cell death. TNFalpha has also been implicated in various caspase-independent cell death processes. Two transcription factors activated by TNFalpha are nuclear factor kappaB (NFkappaB) and activating protein 1 (AP-1). Pathways that promote the activation of these transcription factors involve signaling molecules such as kinases, phospholipases, and sphingomyelinases. In addition to increased survival (anti-apoptotic) gene expression, NFkappaB and AP-1 also induce the expression of genes involved in inflammation, cell growth, and signal regulation. The past decade has witnessed the identification of numerous signaling intermediates implicated in TNFalpha cellular responses. This article reviews the molecular mechanisms of TNFalpha signal transduction. In particular, pathways involved in cell death and transcription factor activation are discussed. PMID- 11005255 TI - Apolipoprotein E and Alzheimer disease: an update on genetic and functional analyses. AB - Exceptional advances have been made in understanding the genetics of how common polymorphisms of the apolipoprotein E gene influence the risk and age of onset of Alzheimer disease (AD). The major genetic susceptibility locus for the common forms of AD, there are 3 common alleles, designated epsilon2, epsilon3, and epsilon4. The inheritance of each dose of APOE4 increases the risk of disease and decreases the age of onset; conversely, the APOE2 allele appears to be protective, by lowering the risk of disease and increasing the age of onset. Testing for the APOE4 allele can be a clinically useful tool in the early diagnosis of cognitively impaired patients suspected of having AD. The APOE4 allele also negatively influences functional recovery following a variety of brain insults. What remains in the study of apolipoprotein E is an explanation of how minor changes in a protein can produce such striking differences in risk and age of onset. In vitro and animal model studies strongly suggest that brain apolipoprotein E is a multifunctional molecule, with potential roles in amyloid deposition and clearance, microtubule stability, intracellular signaling, immune modulation, glucose metabolism, oxidative stress, and other cellular processes. While the relevance of these proposed functions to the etiology of AD remains a mystery, these and other hypotheses will be tested as the field of apoE neurobiology grows, adding relevant new data to the functions of apoE in health and in the pathogenic mechanisms leading to AD. PMID- 11005256 TI - Loss of neurofibromin is associated with activation of RAS/MAPK and PI3-K/AKT signaling in a neurofibromatosis 1 astrocytoma. AB - Neurofibromatosis 1 (NF1) is a common autosomal dominant cancer predisposition syndrome, in which 15% to 20% of affected individuals develop astrocytomas. Neurofibromin, the protein product of the NF1 gene, functions as a tumor suppressor, largely by inhibiting Ras activity. While loss of neurofibromin has been implicated in the molecular pathogenesis of other NF1-associated tumors, there is no formal evidence demonstrating loss of neurofibromin function in NF1 associated astrocytomas. In this report, we describe an NF1 patient from whom both astrocytoma tumor tissue as well as corresponding non-neoplastic white matter were available for analysis. Loss of neurofibromin expression was observed in the tumor and was associated with elevated levels of Ras-GTP. However, elevated Ras-GTP levels were not the result of oncogenic Ras mutations, altered p120-GAP function, growth factor receptor activation, or abnormal p53, Rb, or p16 expression. Furthermore, increased Raf-MAPK and PI3-K/Akt activity was detected in the NF1 astrocytoma compared with the corresponding normal white matter. These results support a role for neurofibromin as the critical GAP in the molecular pathogenesis of NF1 astrocytomas. PMID- 11005257 TI - Cerebral beta amyloid angiopathy is a risk factor for cerebral ischemic infarction. A case control study in human brain biopsies. AB - Cerebral amyloid angiopathy (CAA) is conspicuous for its association with Alzheimer disease (AD) and as a cause of lobar hemorrhages in the elderly, but its role in cerebral infarction is less clear. There is evidence that CAA may also be a risk factor for ischemic infarction in AD. To further investigate CAA as a risk factor for infarction, we studied 108 cases of recent cerebral or cerebellar infarction diagnosed in tissue samples obtained from surgical material. There were 69 males and 39 females with a mean age of 52 yr (range 1 86). The majority of biopsies were obtained from the frontal and parietal lobes. Radiological studies demonstrated a lesion confined to a vascular distribution in 12 of the 17 (71%) cases examined. Microscopic sections stained with hematoxylin and eosin revealed complete, organizing infarction in 107 cases with areas of coagulative necrosis, anoxic-ischemic neuronal injury, inflammation, macrophages, vascular proliferation, gliosis, and swollen axons. One case showed an incomplete infarct. Most cases also exhibited a minor hemorrhagic component with hemosiderin and hematoidin pigments. CAA, defined as amyloid deposition in the walls of leptomeningeal and parenchymal arteries, was found by immunohistochemical stains for beta amyloid in 14 (13%) cases of complete cerebral infarct. Cortical beta amyloid plaques were found by immunohistochemistry in 19 (17%) cases. Cerebral or cerebellar tissues containing cortex and leptomeninges obtained from 136 patients with a mean age of 52 yr (range 1-85) during surgical procedures for diagnosis of primary or metastatic neoplasms and demyelinating lesions were used as age matched controls. In this control group, CAA was found in 5 (3.7%) and beta amyloid plaques in 19 (14%). The results indicate that CAA, but not beta amyloid plaque formation, is significantly more common in patients with ischemic cerebral infarction than in age-matched controls with nonvascular lesions (odds ratio 3.8; 95% confidence interval 1.3-10.9; p < 0.01). Our results indicate that CAA is a risk factor for ischemic cerebral infarction in the population studied. PMID- 11005258 TI - Enhanced CD9 expression in the mouse and human brains infected with transmissible spongiform encephalopathies. AB - A tetraspan protein CD9, normally expressed in the myelin sheath of the central and peripheral nervous system, was identified to be up-regulated in mouse brains infected with transmissible spongiform encephalopathy (TSE), by mRNA differential display screening. To elucidate its role in the neurodegeneration process observed in TSE, CD9 expression was examined in the murine disease model and in the human disease materials. Up-regulation of CD9 gene expression in the TSE infected mouse brains was detected as early as a preclinical stage, when abnormal prion protein deposition and vacuolation were obviously manifested in the internal capsule and thalamus. In contrast, other myelin protein genes showed a reverse pattern of CD9 gene expression. Enhanced CD9 expression was immunohistochemically detected in the astrocytes of such pathological regions. In human specimens of TSE, enhanced CD9 immunoreactivity was observed in the astrocytes and some oligodendrocytes in the brains, but no relevant alteration in CD9 immunoreactivity was observed in the other organs or tissues. Positive CD9 immunoreactivity in astrocytes was also manifest in other neurological disorders in a less prominent manner. The findings indicate that up-regulated CD9 plays a role in glial cells in pathological conditions, especially in such a devastating condition as TSE. PMID- 11005259 TI - Disulfiram produces a non-carbon disulfide-dependent schwannopathy in the rat. AB - Disulfiram is a dithiocarbamate drug used for alcohol aversion therapy that produces a distal sensorimotor peripheral neuropathy in certain individuals. Because carbon disulfide, a disulfiram metabolite, produces a peripheral neuropathy clinically similar to disulfiram, it has been postulated that disulfiram neuropathy results from CS2 release in vivo. The current study evaluated the morphological changes produced by disulfiram and the contribution of CS2-mediated protein cross-linking to disulfiram-induced neuropathy. Male Sprague-Dawley rats were administered 1% w/w disulfiram in their feed for 2, 4, 5, or 7 wk, and erythrocyte spectrin, hemoglobin, and neurofilament preparations were isolated and the extent of cross-linking assessed by SDS-PAGE, RP-HPLC, and Western blotting, respectively. Spinal cord and peripheral nerve sections were obtained from separate treated animals and assessed by light and electron microscopy. Significant protein cross-linking was only detected in neurofilament preparations obtained after 7 wk of exposure. Morphological changes were observed after 4 wk exposure and consisted of vacuoles within the Schwann cell cytoplasm and segmental demyelination. No neurofilamentous axonal swellings were detected and no significant changes were observed in the CNS. Because disulfiram neuropathy lacks both the morphological changes and intermolecular cross-linking characteristic of CS2, we conclude that disulfiram neuropathy is not mediated by the axonal toxicant CS2; instead, disulfiram appears to be a primary Schwann cell toxicant. Recognition of a diethylcarbamoyl adduct on globin and axonal proteins presents a novel putative neurotoxic mechanism for disulfiram. PMID- 11005260 TI - Nitric oxide synthase activity and expression in experimental diabetic neuropathy. AB - The changes of nitric oxide synthase (NOS) activity and expression in experimental diabetic neuropathy have not been examined. Increases in ganglia NOS might be similar to those that follow axotomy, whereas declines in endothelial NOS (eNOS) and immunological NOS (iNOS) might explain dysfunction of microvessels or macrophages. In this work, we studied NOS activity in lumbar dorsal root ganglia (DRG) of rats with both short- and long-term experimental streptozotocin induced diabetes and correlated it with expression of each of the 3 NOS isoforms. NOS enzymatic activity in DRG increased after 12 months of diabetes. This increase, however, was not accompanied by an increase in neuronal NOS immunohistochemistry or mRNA. Immunohistochemical and RT-PCR studies did not identify changes of eNOS expression in 12-month sciatic nerves or DRG from diabetics. Two-month diabetic DRG had increased eNOS mRNA and there was novel eNOS labeling of capsular DRG and perineurial cells. iNOS mRNA levels were lower in diabetics at both time points in peripheral nerves but were unchanged in DRG. Diabetic ganglia showed an increase in NOS activity not explained by novel NOS isoform synthesis. The increases may compensate for NO "quenching" by endproducts of glycosylation. Declines in iNOS may indicate impaired macrophage function. PMID- 11005261 TI - Further evidence for the involvement of epidermal growth factor in the signaling pathway of vitamin B12 (cobalamin) in the rat central nervous system. AB - In order to get further evidence for a mandatory involvement of epidermal growth factor (EGF) in the neutrophic action of vitamin B12 (cobalamin (Cbl)) in the central nervous system (CNS) of the rat, we observed the effects of repeated intracerebroventricular (ICV) microinjections of EGF in rats made Cbl-deficient through total gastrectomy. Morphometric analysis demonstrated a significant reduction in both intramyelinic and interstitial edema in the white matter of the spinal cord (SC) of totally gastrectomized (TGX) rats after treatment. Intramyelinic and interstitial edema are characteristic of Cbl-deficient central neuropathy in the rat. Similar lesions were also present in SC white matter of rats treated with repeated ICV microinjections of specific anti-EGF antibodies without any modification in their Cbl status. These results, together with those of a previous study showing the cessation of EGF synthesis in the CNS of TGX rats, demonstrate that: a) EGF is necessarily involved in the signaling pathway of Cbl in the rat CNS; and b) the lack of a neurotrophic growth factor EGF, and not the mere withdrawal of Cbl, causes or at least contributes to neurodegenerative Cbl-deficient central neuropathy. PMID- 11005262 TI - Comparative genomic hybridization in pineal germ cell tumors. AB - Fifteen primary pineal germ cell tumors (8 germinomas, 4 mixed teratomas germinomas, 2 immature teratomas, and 1 yolk sac tumor) and 2 recurrences of the yolk sac tumor were studied by comparative genomic hybridization (CGH). An average of 1.8 chromosomal changes per germinoma (0.5 gains vs 1.3 losses), 5.5 per mixed teratoma-germinoma (3.0 gains vs 2.5 losses), 3.5 per immature teratoma (2.0 gains vs 1.5 losses), and 2.0 in the yolk sac tumor (2 gains vs 0 losses) were found; the first recurrence showed 7 (4 gains vs 3 losses), the second 13 imbalances (8 gains vs 5 losses). The most frequent imbalances were gains on 12p (40%), 8q (27%), and 1q (20%) as well as losses on 13q (47%), 18q (33%), 9q and 11q (20% each). Among germinomas, the most common chromosomal changes were -13q and -18q (38% each), in mixed teratomas-germinomas +8q (100%), +12p (75%), -13q (75%) and -9q (50%). Seven high-level gains were identified: 5 in mixed teratomas germinomas (+8q: 3 cases, + 12p: 2 cases), 1 each in a germinoma (+2p) and an immature teratoma (+12p). Minimal common regions of over- and underrepresentation were found on +8q11.22-21.1, +12p11.1-12.1, -9q32-qter, -11q23.2-qter, -13q32 qter and -18q22-qter. Our findings suggest, that imbalances in cerebral germ cell tumors affect the same chromosomes as among their extracerebral counterparts, albeit in a considerably lower frequency among cerebral germinomas where +12p does not seem to play a major role. PMID- 11005263 TI - Tracing cranial nerve pathways (glossopharyngeal, vagus, and hypoglossal) in SIDS and control infants: a DiI study. AB - It has been proposed that Sudden Infant Death Syndrome (SIDS) might occur as a consequence of a developmental deficit associated with the cardiorespiratory and arousal control centers located within the brainstem. In this study 1.1' dioctadecyl-3,3,3',3-tetramethylindocarbocyanine perchlorate (DiI) was used to investigate the trajectories of the glossopharyngeal and vagus nerves which carry essential afferent and efferent fiber tracts associated with cardiac and respiratory control and of the hypoglossal nerve which innervates the tongue, in SIDS (n = 14) and control (n = 7) infants. The postnatal development of the trajectories of these nerves was examined in non-SIDS brains and comparisons were then made with age-matched SIDS brains. The mean profile area of hypoglossal and dorsal motor neurons were also assessed. In controls, no major alterations were observed in the trajectories of axon bundles with increasing age (7 wk to 2 yr) in each of the nerves investigated although axon bundles appeared to increase in thickness with age. In SIDS cases (2 wk to 44 wk), the trajectories of the cranial nerves were not different from those seen in age-matched control cases. The mean profile area of hypoglossal and dorsal motor neurons was not significantly different between control and SIDS infants. We conclude that the DiI tracing technique can be used successfully to trace the pathways of cranial nerves in human infant fixed-tissue. Furthermore, if functional differences exist between SIDS and non-SIDS brains in the control of respiration, circulation, or arousal they do not appear to be related to markedly reduced or aberrant projections of the glossopharyngeal, vagus, or hypoglossal nerves. PMID- 11005264 TI - Immunohistochemical and biochemical studies demonstrate a distinct profile of alpha-synuclein permutations in multiple system atrophy. AB - Although alpha-synuclein (alpha-syn) has been implicated as a major component of the abnormal filaments that form glial cytoplasmic inclusions (GCIs) in multiple system atrophy (MSA), it is uncertain if GCIs are homogenous and contain full length alpha-syn. Since this has implications for hypotheses about the pathogenesis of GCIs, we used a novel panel of antibodies to defined regions throughout alpha-syn in immunohistochemical epitope mapping studies of GCIs in MSA brains. Although the immunostaining profile of GCIs with these antibodies was similar for all MSA brains, there were significant differences in the immunoreactivity of the alpha-syn epitopes detected in GCIs. Notably, carboxy terminal alpha-syn epitopes were immunodominant in GCIs, but the entire panel of antibodies immunostained cortical Lewy bodies (LBs) in dementia with LBs brain with similar intensity. While the distribution of alpha-syn labeled GCIs paralleled that previously reported using silver stains, antibodies to carboxy terminal alpha-syn epitopes revealed a previously undescribed burden of GCIs in the MSA hippocampal formation. Finally, Western blots demonstrated detergent insoluble monomeric and high-molecular weight alpha-syn species in GCI rich MSA cerebellar white matter. Collectively, these data indicate that alpha-syn is a prominent component of GCIs in MSA, and that GCIs and LBs may result from cell type specific conformational or post-translational permutations in alpha-syn. PMID- 11005265 TI - Evidence for a positive correlation between serum cortisol levels and IL-1beta production by peripheral mononuclear cells in anorexia nervosa. AB - A hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis has been reported in anorexia nervosa (AN), together with some immunological abnormalities, involving citokine - and particularly Tumor Necrosis-Factor-alpha (TNF-alpha) - production by polymorphonuclear cells. The ability of pro inflammatory cytokines to activate the HPA axis is well known; however, there are no data demonstrating an interdependence between immunological and endocrine response in AN. To investigate the presence of a correlation between immune response and pituitary-adrenal function, plasma ACTH and serum cortisol concentrations were measured in 13 AN patients and in the same number of controls. TNF-alpha and interleukin (IL)-1beta production by ex-vivo unstimulated and LPS-stimulated peripheral mononuclear cells was also assessed. Circulating cortisol concentrations were higher (p<0.01) in AN (156.7 +/- 45.1 microg/l, mean +/- SD) than in controls (105.9 +/- 25.7 microg/l). Unstimulated IL-1beta release in supernatants of mononuclear cell cultures was slightly but not significantly higher in AN than in controls, while TNF-alpha release was similar in the two groups. A positive correlation was found between IL-1beta concentrations in unstimulated culture supranatants and serum cortisol levels in AN (r=0.782, p=0.002), while in normal subjects there was a trend toward a negative correlation; a slight positive correlation, while not significant, between IL 1beta and plasma ACTH, as well as between TNF-alpha and serum cortisol was also found in AN. These data suggest that the normal relationship between pro inflammatory cytokines release, particularly IL-1beta, and cortisol secretion is deranged in AN. PMID- 11005266 TI - A randomized cross-over study comparing cabergoline and quinagolide in the treatment of hyperprolactinemic patients. AB - Quinagolide (QUI) and cabergoline (CAB) are dopamine agonists recently introduced for the treatment of hyperprolactinemia. In the present study, these drugs have been compared in terms of effectiveness and tolerability. Twenty patients (18 females and 2 males) with hyperprolactinemia (8 with microprolactinomas, 6 with idiopathic hyperprolactinemia and 6 with empty sella turcica syndrome) were treated with oral QUI (75 microg once daily) and CAB (0,5 mg twice weekly), in a randomized cross-over trial with placebo between both drugs. Each drug was administered for 12 weeks, separated by other 12 weeks with placebo. PRL levels decreased with both drugs at 2 or 4 weeks of starting the treatment, without differences between both drugs at weeks 4, 8 and 12. At week 12, normal PRL levels (<20 ng/ml) were attained in 90% patients with CAB and only in 75% patients with QUI (p<0.05). After discontinuation of treatment, significant increase in serum PRL was higher after QUI withdrawal than after CAB. Clinical efficacy of both treatments was similar in terms of improvement amenorrhea, oligomenorrhea, galactorrhea, and impotence. All patients completed both cycles of treatment, and the most frequent side-effects were nausea, headache and dizziness, without significant differences between CAB (30%) and QUI (55%). Our study indicates that, at the doses employed here, CAB showed a high percentage of patients with normal PRL at the end of treatment and long-lasting efficacy in the levels of PRL. Clinical response and side-effects were similar in both drugs. PMID- 11005267 TI - Urinary cortisol to cortisone metabolites in hypertensive obese children. AB - Childhood obesity is accompanied by a variety of cardiovascular risk factors (hypertension, insulin resistance, dyslipidaemia) which tend to aggregate (syndrome X). 11beta-hydroxysteroid dehydrogenase (11beta-HSD) is supposed to play a role in the pathogenesis of hypertension and the development of syndrome X. There are two isoforms of 11beta-HSD. 11beta-HSD-2 is responsible for the inactivation of cortisol to inactive cortisone. In the case of impaired enzyme activity the ratio of urinary tetrahydrocortisol (THF)+ its isomer allotetrahydrocortisol (5alpha-THF)/tetrahydrocortisone (THE) is elevated. 11beta HSD-1 is an oxo-reductase, which type catalyses the conversion of cortisone to cortisol. The aim of the present study was to investigate if there was any alteration in the urinary cortisol metabolites reflecting 11beta-HSD activity in hypertensive obese children (no.=15) as compared to normotensive obese (no.=11) and normotensive non-obese children (no.=15). We found an increased excretion of cortisol metabolites in hypertensive obese children compared to obese and normal weight children having normal blood pressure. The ratio of THF+5alpha(THF/THE had a significant correlation with systolic blood pressure. On the basis of our study the ratio of THF+5alpha-THF/ THE reflecting on altered enzyme activity seems to be an independent factor influencing especially systolic blood pressure in hypertensive obese children. PMID- 11005268 TI - Sexual dimorphism in the hypothalamo-pituitary-adrenal (HPA) axis and TNFalpha responses to phospholipase A2-related neurotoxin (from crotalus durissus terrifcus) challenge. AB - Neuroendocrine-immune interactions are vital for the individual's survival in certain physiopathological conditions such as sepsis and tissular injury. It is known that several snake venoms (SV) are potent neurotoxic compounds and that their main component is a specific phospholipase type 2 (PLA2). It has been recently described that the venom from crotalus durissus terrificus (SV) possesses a cytotoxic effect in different in vitro and in vivo animal models. In the present study we investigated whether SV is able to stimulate both TNFalpha and neuroendocrine functions in a sexual dimorphic fashion. For this purpose the modulatory role of endogenous sex steroids during neurotoxemia was evaluated. Our results indicate that SV (25 microg/animal) stimulates the hypothalamo-pituitary adrenal axis and TNFalpha secretion when administered (ip) to adult male mice, such responses were characterized by a time-related enhance in plasma glucose, ACTH, corticosterone and TNFalpha levels. SV-stimulated glycemia, corticosteronemia and adrenal glucocorticoid were sexually dimorphic. Twenty-day gonadectomized mice showed a similar sexual dimorphism to that found in intact animals, however, they additionally showed a sexual dimorphic pattern in cytokine release in plasma 30 min post-SV. Estradiol (E2) treatment, in gonadectomized mice, abolished some characteristics of the sexual dimorphism, such as hyperglycemia, hypercorticosteronemia and hypercytokinemia. Finally, in vitro experiments indicate that: a) gonadectomy increased spontaneous and SV-stimulated cytokine output by incubated peripheral mononuclear cells (PMNC), regardless of the sex; and b) despite E2 treatment, in gonadectomized, did not modify the pattern of basal and SV-elicited TNFalpha secretion induced by orchidectomy, fully reversed the enhance in basal and SV-stimulated cytokine release found after ovariectomy alone. Our results further indicate that neurotoxemia, due to SV challenge, induces several symptoms common to those of inflammatory stress; they also strongly support that both gender and endogenous sex steroids are responsible for neuroendocrine-immunological sexual dimorphism. PMID- 11005269 TI - Intersubject responsiveness of high-affinity growth hormone (GH)-binding protein (GHBP) to long-term GH replacement therapy. AB - In adult growth hormone deficiency (GHD) syndrome responsiveness to GH replacement therapy is reported to vary considerably. The underlying mechanisms, however, are not well understood. The aim of this study was to investigate which baseline variables determine the reported variable intersubject responsiveness of high-affinity GH-binding protein (GHBP) to GH replacement therapy. In the setting of a double blind study over 12 months with placebo control over the first 6 months, we analyzed the interrelationship between a number of baseline variables, which vary considerably amongst subjects, and the GHBP response to GH replacement in 31 GHD adults (21 males and 10 females). The following variables were investigated: age, gender, duration of GHD, body composition, serum levels of high-affinity GHBP, insulin-like growth factor-1 (IGF-1), and IGF-binding protein 3 (IGFBP-3). The results showed that in the 6 months treated group of 16 patients (11 males, 5 females), serum IGF-1 increased from 87 ng/ml (range: 26 to 173) to 250 (range: 62 to 467) (p<0.01) and GHBP increased from 1,302 pmol/l (range: 845 to 1,m960) to 1418 (range: 941 to 2,025) (p=0.04). Both parameters showed a significant time effect (within-subjects) (p<0.001). In the 12 months treated group of 15 patients (10 males, 5 females), serum IGF-1 increased from 92 ng/ml (range: 20 to 180) to 272 (range: 45 to 491) (p<0.01), whereas GHBP did not show a significant change: from 1,186 pmol/l (range: 660 to 1,690) to 1,252 (range: 580 to 1,890) (p=0.87). Also no significant time effect (within-subjects) was observed for GHBP (p=0.06). Step-wise multiple regression analyses revealed that during the 6 months placebo period baseline GHBP explained 83% of the variance in post-placebo GHBP, whereas the variance in post-treatment GHBP could be accurately predicted (adjusted R2=0.93) from baseline GHBP and body fat mass, irrespective of the duration of GH treatment. No other baseline variables contributed independently to the GHBP response, with the exception of IGFBP-3, which showed a small, but significant contribution in females, but not in males. These findings indicate that the variable intersubject responsiveness of GHBP to GH replacement therapy is mainly due to differences in baseline body fat mass amongst adult GHD patients, and that in female patients a relatively low baseline IGFBP-3 contributes to a rise in serum GHBP after GH treatment. The clinical relevance of measuring GHBP in adult GHD patients is limited to the first screening step to diagnose GHD, because long-term GH therapy tends to restore serum GHBP to pretreatment levels. PMID- 11005270 TI - Does kidney transplantation normalise cortisol metabolism in apparent mineralocorticoid excess syndrome? AB - The syndrome of apparent mineralocorticoid syndrome (AME) results from defective 11beta-hydroxysteroid dehydrogenase 2 (11beta-HSD2). This enzyme is co-expressed with the mineralocorticoid receptor (MR) in the kidney and converts cortisol to its inactive metabolite cortisone. Its deficiency allows the unmetabolized cortisol to bind to the MR inducing sodium retention, suppression of PRA and hypertension. Thus, the syndrome is a disorder of the kidney. We present here the first patient affected by AME cured by kidney transplantation. Formerly, she was considered to have a mild form of the syndrome (Type II), but progressively she developed renal failure which required dialysis and subsequent kidney transplantation. To test the ability of the transplanted kidney to normalise the patient's cortisol metabolism, we gave, in two different experiments, 25 and 50 mg/day of cortisone acetate or 15 and 30 mg/day of cortisol after inhibition of the endogenous cortisol by synthetic glucocorticoid (methylprednisolone and dexamethasone). The AME diagnostic urinary steroid ratios tetrahydrocortisol+5alphatetrahydrocortisol/tetrahydrocortisone and cortisol/cortisone were measured by gas chromatography/mass spectrometry. Transplantation resulted in lowering blood pressure and in normalization of serum K and PRA. After administration of a physiological dose of cortisol (15 mg/day), the urinary free cortisol/cortisone ratio was corrected (in contrast to the A ring reduced metabolites ratio), confirming that the new kidney had functional 11beta-HSD2. This ratio was abnormally high when the supra-physiological dose of cortisol 30 mg/day was given. After cortisone administration, the tetrahydrocortisol+5alphatetrahydrocortisol/tetrahydrocortisone ratio resulted normalised with both physiological and supra-physiological doses, confirming that the hepatic reductase activity is not affected. As expected, the urinary free cortisol/cortisone ratio was normal with physiological, but increased after supra physiological doses of cortisone. The described case indicates a normalisation of cortisol metabolism after kidney transplantation in AME patient and confirms the supposed pathophysiology of the syndrome. Moreover, it suggests a new therapeutic strategy in particularly vulnerable cohorts of patients inadequately responsive to drug therapy or with kidney failure. PMID- 11005271 TI - Cabergoline modulation of alpha-subunits and FSH secretion in a gonadotroph adenoma. AB - Most non-functioning pituitary adenomas respond poorly to medical therapy. We describe the case of a 62-year-old man who presented with clinical features of an invasive macroadenoma. Baseline hormonal evaluation revealed increased FSH and alpha-subunit (alpha-SU) levels. Transsphenoidal exeresis followed by radiotherapy (RT) was performed. Almost all neoplastic cells were intensely immunoreactive for alpha-SU. On PCR analysis, specific amplification products were observed for somatostatin 2, 3 and 5 receptors as well as for both short and long isoforms of the dopamine D2 receptor. In vitro, alpha-SU and FSH were released into the medium by adenoma cells and increased after TRH stimulation. After surgery, alpha-SU and FSH levels were still elevated. Short-term slow release lanreotide treatment did not modify either alpha-SU or FSH levels. Cabergoline was started and a fast and long-lasting decrease in alpha-SU and, to a lesser extent, in FSH was observed. The tumor remnant was unmodified on magnetic resonance imaging 3 years after surgery and RT. This case report shows that the in vitro expression of somatostatin receptors may not be directly associated to the in vivo response of alpha-SU and FSH to lanreotide, probably because of a functional uncoupling of the receptors. Cabergoline should be considered as an effective therapy for hormonal, and perhaps proliferative, control of gonadotroph adenoma remnants before the effects of RT are fully effective. PMID- 11005272 TI - Extremely high levels of estradiol and testosterone in a case of polycystic ovarian syndrome. Hormone and clinical similarities with the phenotype of the alpha estrogen receptor null mice. AB - A 19-year-old nulliparous hirsute woman was evaluated for the very high serum levels of testosterone (T) and estradiol (E2) measured in an outside laboratory. Menarche had occurred at 11 years and was followed by regular menses. We confirmed the high levels of T (9-16 ng/ml, nv 0.2-0.8) and E2 (>1,000 pg/ml, nv 30-120). LH and FSH were consistently high (73-118 mU/l and 18-29 mU/l, respectively; LH/FSH ratio=4.1-4.7) and responsive to iv GnRH (LH baseline=118 mU/I, 30 min=290; FSH baseline=25 mU/l, 30 min=46). The unstimulated values contrasted with those (LH=12, FSH=8 mU/I) measured in the outside laboratory, suggesting antigenically anomalous gonadotropins. 17-OH-progesterone was normal (0.5 ng/ml). After 1 mg dexamethasone, serum cortisol was normally suppressed (24 ->0.4 microg/dl), T declined minimally (9-->8.6 ng/ml) and E2 remained high (>1,000 pg/ml). An exploratory laparotomy was performed, and two enlarged ovaries with multiple cysts as in a typical polycystic ovarian syndrome (PCOS) were seen. Before the wedge resection of the ovaries, hormones were assayed in the ovary veins (right ovary: T=30 ng/ml, Pg=17 ng/ml, E2=>5,000 pg/ml; left: T=14 ng/ml, Pg=14 ng/ml, E2=>5,000 pg/ml). Histologically, the follicle cysts showed luteinization of the theca interna; there was no evidence for ovary tumor in either ovary. After 21 days of 35 microg ethynyl-E2+2 mg cyproterone acetate (CA), E2=3,000 pg/ml, T=1.4 ng/ml, LH=10.5 mU/l and FSH=4.1 mU/I. After three cycles of the said therapy (but with 50 mg CA in the first 10 days of each cycle), E2 was 1,600 pg/ml, T 1.7 ng/ml, LH 7.1 and FSH 4.6 mU/I. Based on similarities with the phenotype of the alpha estrogen receptor knockout female mice (alphaERKO), one possible explanation for the puzzling clinical and biochemical picture of our patient is resistance of (alphaER to estrogens. This is the first case of PCOS with extremely high E2 and T. Thus, the differential diagnosis of high levels of E2 +/- T should include PCOS. PMID- 11005273 TI - The use of recombinant human TSH in a patient with metastatic follicular carcinoma and insufficient endogenous TSH production. AB - We present a case of a patient suffering from metastatic differentiated thyroid carcinoma (DTC) and insufficient endogenous TSH production suspicious of secondary hypothyroidism. The use of recombinant human TSH (rhTSH) enabled us to administer a therapeutic activity of radioactive iodine (RAI) under maximal TSH stimulation, achieving a marked decrease in thyroglobulin accompanied by a clinical improvement. PMID- 11005274 TI - Adrenal bilateral incidentaloma by reactivated histoplasmosis. AB - We report a case of bilateral adrenal incidentaloma caused by the capsulatum variety of Histoplasma capsulatum diagnosed in a 74 years old man born in and a life time resident of Treviso, Italy, with the exception of two years spent in Pakistan (1964-1966) as a well-driller. The patient was hospitalized in 1995 for alcoholic chronic hepatitis, chronic Helicobacter pylori gastritis and post infarction ischemic cardiomyopathy. Abdominal ultrasound incidentally showed bilateral adrenal masses (the right one 6.3 cm in diameter) confirmed by computed tomography, with adrenal function within normal limits. After three months, the patient was again hospitalized due to evening fever, asthenia, anorexia, weight loss and occasional hyperhidrosis. Abdominal ultrasound showed an increase of the right adrenal lesion with normal adrenal function. Ultrasound-guided fine needle aspiration did not prove useful for diagnosis. Accordingly, a laparotomy with bilateral biopsy was performed; histology showed the presence of numerous tissue form cells of H. capsulatum variety capsulatum. Serum anti-H. capsulatum antibodies were negative. Since March, 1996, the patient was given itraconazole and his symptoms quickly regressed but the computed tomography findings, however, have not changed and the patient has adrenal hypofunction that is being treated with cortisone acetate. PMID- 11005275 TI - Reflections on growth hormone. PMID- 11005277 TI - Hyperprolactinemia preceding Cushing's disease. PMID- 11005276 TI - The metabolic effects of cyclosporin and tacrolimus. AB - The introduction of cyclosporin and, more recently, tacrolimus in the immunosuppression of transplanted patients has lead to prolonged graft survival and increased patients' life expectancy. It has been therefore possible to evaluate the effects of long-term treatment with these drugs and metabolic alterations in patients on cyclosporin or tacrolimus have been reported by several authors. In particular, the use of these drugs is associated with abnormalities of glucose and lipid metabolism. Post-transplant diabetes is more common with tacrolimus, probably due to more marked effects on the pancreatic beta-cells, whereas increased levels of cholesterol and triglycerides are more frequently associated with cyclosporin treatment, even though, in this latter case, steroid treatment seems to play a major role. Comparison and intervention studies must be planned to evaluate the best therapeutical approaches to control these abnormalities and to assess the possibility to further increase graft and patient survival by appropriate treatment of diabetes and hyperlipidemia. PMID- 11005278 TI - Evolutionary developmental biology and the problem of variation. AB - One of the oldest problems in evolutionary biology remains largely unsolved. Which mutations generate evolutionarily relevant phenotypic variation? What kinds of molecular changes do they entail? What are the phenotypic magnitudes, frequencies of origin, and pleiotropic effects of such mutations? How is the genome constructed to allow the observed abundance of phenotypic diversity? Historically, the neo-Darwinian synthesizers stressed the predominance of micromutations in evolution, whereas others noted the similarities between some dramatic mutations and evolutionary transitions to argue for macromutationism. Arguments on both sides have been biased by misconceptions of the developmental effects of mutations. For example, the traditional view that mutations of important developmental genes always have large pleiotropic effects can now be seen to be a conclusion drawn from observations of a small class of mutations with dramatic effects. It is possible that some mutations, for example, those in cis-regulatory DNA, have few or no pleiotropic effects and may be the predominant source of morphological evolution. In contrast, mutations causing dramatic phenotypic effects, although superficially similar to hypothesized evolutionary transitions, are unlikely to fairly represent the true path of evolution. Recent developmental studies of gene function provide a new way of conceptualizing and studying variation that contrasts with the traditional genetic view that was incorporated into neo-Darwinian theory and population genetics. This new approach in developmental biology is as important for microevolutionary studies as the actual results from recent evolutionary developmental studies. In particular, this approach will assist in the task of identifying the specific mutations generating phenotypic variation and elucidating how they alter gene function. These data will provide the current missing link between molecular and phenotypic variation in natural populations. PMID- 11005279 TI - The effects of subdivision on the genetic divergence of populations and species. AB - An island model of migration is used to study the effects of subdivision within populations and species on sample genealogies and on between-population or between-species measures of genetic variation. The model assumes that the number of demes within each population or species is large. When populations (or species), connected either by gene flow or historical association, are themselves subdivided into demes, changes in the migration rate among demes alter both the structure of genealogies and the time scale of the coalescent process. The time scale of the coalescent is related to the effective size of the population, which depends on the migration rate among demes. When the migration rate among demes within populations is low, isolation (or speciation) events seem more recent and migration rates among populations seem higher because the effective size of each population is increased. This affects the probability of reciprocal monophyly of two samples, the chance that a gene tree of a sample matches the species tree, and relative likelihoods of different types of polymorphic sites. It can also have a profound effect on the estimation of divergence times. PMID- 11005280 TI - Coevolutionary clines across selection mosaics. AB - Much of the dynamics of coevolution may be driven by the interplay between geographic variation in reciprocal selection (selection mosaics) and the homogenizing action of gene flow. We develop a genetic model of geographically structured coevolution in which gene flow links coevolving communities that may differ in both the direction and magnitude of reciprocal selection. The results show that geographically structured coevolution may lead to allele-frequency clines within both interacting species when fitnesses are spatially uniform or spatially heterogeneous. Furthermore, the results show that the behavior and shape of clines differ dramatically among different types of coevolutionary interaction. Antagonistic interactions produce dynamic clines that change shape rapidly through time, producing shifting patterns of local adaptation and maladaptation. Unlike antagonistic interactions, mutualisms generate stable equilibrium patterns that lead to fixed spatial patterns of adaptation. Interactions that vary between mutualism and antagonism produce both equilibrium and dynamic clines. Furthermore, the results demonstrate that these interactions may allow mutualisms to persist throughout the geographic range of an interaction, despite pockets of locally antagonistic selection. In all cases, the coevolved spatial patterns of allele frequencies are sensitive to the relative contributions of gene flow, selection, and overall habitat size, indicating that the appropriate scale for studies of geographically structured coevolution depends on the relative contributions of each of these factors. PMID- 11005281 TI - Fixation of clonal lineages under Muller's ratchet. AB - For clonal lineages of finite size that differ in their deleterious mutational effects, the probability of fixation is investigated by mathematical theory and Monte Carlo simulations. If these fitness effects are sufficiently small in one or both lineages, then the lineage with the less deleterious effects will become fixed with high probability. If, however, in both lineages the deleterious effects are larger than a threshold s(c), then the probability of fixation is independent of the fitness effects and depends only on the initial frequencies of the lineages. This threshold decreases with decreasing genomic mutation rate U and increases with population size N. (For N = 10(5), we have s(c) approximately = 0.1 if U = 1, and s(c) approximately = 0.015 if U = 0.1). Above the threshold, the competition is not driven by the ratio of mean fitnesses of the lineages, but by the relative sizes of the zero-mutation classes, which are independent of the fitness effects of the mutations. After the loss of the zero-mutation class of a lineage, the other lineage will spread to fixation with high probability and within a short time span. If the mutation rates of the lineages differ substantially, the lineage with the lower mutation rate is fixed with very high probability unless the lineage with the larger mutation rate has very slightly deleterious mutational effects. If the mutation rates differ by not more than a few percent, then the lineage with the higher mutation rate and the more deleterious effects can become fixed with appreciable probability for a certain range of parameters. The independence of the fixation probability on the fitness effects in a single population leads to dramatic effects in metapopulations: lineages with more deleterious effects have a much higher fixation probability. The critical value s(c), above which this phenomenon occurs, decreases as the migration rate between the subpopulations decreases. PMID- 11005282 TI - Patterns of parapatric speciation. AB - Geographic variation may ultimately lead to the splitting of a subdivided population into reproductively isolated units in spite of migration. Here, we consider how the waiting time until the first split and its location depend on different evolutionary factors including mutation, migration, random genetic drift, genetic architecture, and the geometric structure of the habitat. We perform large-scale, individual-based simulations using a simple model of reproductive isolation based on a classical view that reproductive isolation evolves as a by-product of genetic divergence. We show that rapid parapatric speciation on the time scale of a few hundred to a few thousand generations is plausible even when neighboring subpopulations exchange several individuals each generation. Divergent selection for local adaptation is not required for rapid speciation. Our results substantiates the claims that species with smaller range sizes (which are characterized by smaller local densities and reduced dispersal ability) should have higher speciation rates. If mutation rate is small, local abundances are low, or substantial genetic changes are required for reproductive isolation, then central populations should be the place where most splits take place. With high mutation rates, high local densities, or with moderate genetic changes sufficient for reproductive isolation, speciation events are expected to involve mainly peripheral populations. PMID- 11005283 TI - Overlapping generations can promote altruistic behavior. AB - We use an inclusive fitness model to study the evolution of altruism in a patch structured population in which there is positive probability of breeder survival from one generation to the next. We find first that breeder survival promotes altruism and second that there is a marked difference between benefits of fecundity and benefits of survival. Under the first altruism is more strongly favored, and under the second altruism is less strongly favored than in a randomly mixing population. PMID- 11005284 TI - The coevolution of parasites with host-acquired immunity and the evolution of sex. AB - Here I present a deterministic model of the coevolution of parasites with the acquired immunity of their hosts, a system in which coevolutionary oscillations can be maintained. These dynamics can confer an advantage to sexual reproduction within the parasite population, but the effect is not strong enough to outweigh the twofold cost of sex. The advantage arises primarily because sexual reproduction impedes the response to fluctuating epistasis and not because it facilitates the response to directional selection-in fact, sexual reproduction often slows the response to directional selection. Where the cost of sexual reproduction is small, a polymorphism can be maintained between the sexuals and the asexuals. A polymorphism is maintained in which the advantage gained due to recombination is balanced by the cost of sex. At much higher costs of sex, a polymorphism between the asexual and sexual populations can still be maintained if the asexuals do not have a full complement of genotypes available to them, because the asexuals only outcompete those sexuals with which they share the same selected alleles. However, over time we might expect the asexuals to amass the full array of genotypes, thus permanently eliminating sexuals from the population. The sexuals may avoid this fate if the parasite population is finite. Although the model presented here describes the coevolution of parasites with the acquired immune responses of their hosts, it can be compared with other host parasite models that have more traditionally been used to investigate Red Queen theories of the evolution of sex. PMID- 11005285 TI - Population differentiation in an annual legume: genetic architecture. AB - The presence or absence of epistasis, or gene interaction, is explicitly assumed in many evolutionary models. Although many empirical studies have documented a role of epistasis in population divergence under laboratory conditions, there have been very few attempts at quantifying epistasis in the native environment where natural selection is expected to act. In addition, we have little understanding of the frequency with which epistasis contributes to the evolution of natural populations. In this study we used a quantitative genetic design to quantify the contribution of epistasis to population divergence for fitness components of a native annual legume, Chamaecrista fasciculata. The design incorporated the contrast of performance of F2 and F3 segregating progeny of 18 interpopulation crosses with the F1 and their parents. Crosses were conducted between populations from 100 m to 2000 km apart. All generations were grown for two seasons in the natural environment of one of the parents. The F1 often outperformed the parents. This F1 heterosis reveals population structure and suggests that drift is a major contributor to population differentiation. The F2 generation demonstrated that combining genes from different populations can sometimes have unexpected positive effects. However, the F3 performance indicated that combining genes from different populations decreased vigor beyond that due to the expected loss of heterozygosity. Combined with previous data, our results suggest that both selection and drift contribute to population differentiation that is based on epistatic genetic divergence. Because only the F3 consistently expressed hybrid breakdown, we conclude that the epistasis documented in our study reflects interactions among linked loci. PMID- 11005286 TI - Population differentiation in an annual legume: local adaptation. AB - Studies of many plants species have demonstrated adaptive genetic differentiation to local environmental conditions. Typically these studies are conducted to evaluate adaptation to contrasting environments. As a consequence, although local adaptation has been frequently demonstrated, we have little information as to the spatial scale of adaptive evolution. We evaluated adaptive differentiation between populations of the annual legume Chamaecrista fasciculata using a replicated common-garden design. Study sites were established in three field locations that are home to native populations of C. fasciculata. Each location was planted for two years with seed from the population native to the study site (home population) and populations located six distances (0.1-2000 km) from each site (transplanted populations). Seeds were planted into the study sites with minimum disturbance to determine the scale of local adaptation, as measured by a home-site fitness advantage, for five fitness components: germination, survival, vegetative biomass, fruit production, and the number of fruit produced per seed planted (an estimate of cumulative fitness). For all characters there was little evidence for local adaptation, except at the furthest spatial scales. Patterns of adaptive differentiation were fairly consistent in two of the three sites, but varied between years. Little genetic variation was expressed at the third site. These results, combined with previous estimates of limited gene flow, suggest that metapopulation processes and temporal environmental variation act together to reduce local adaptation, except over long distances. PMID- 11005287 TI - Fitness differences among diploids, tetraploids, and their triploid progeny in Chamerion angustifolium: mechanisms of inviability and implications for polyploid evolution. AB - Theoretical models indicate that the evolution of tetraploids in diploid populations will depend on both the relative fitness of the tetraploid and that of the diploid-tetraploid hybrids. Hybrids are believed to have lower fitness due to imbalances in either the ploidy (endosperm imbalance) or the ratio of maternal to paternal genomes in their endosperm (genomic imprinting). In this study we created diploids, tetraploids, and hybrid triploids of Chamerion angustifolium from crosses between field-collected diploid and tetraploid plants and evaluated them at six life stages in a greenhouse comparison. Diploid offspring (from 2x x 2x crosses) had significantly higher seed production and lower biomass than tetraploid offspring (from 4x x 4x crosses). Relative to the diploid, the cumulative fitness of tetraploids was 0.67. In general, triploids (from 2x x 4x, 4x x 2x crosses) had significantly lower seed production, lower pollen viability, and higher biomass than diploid individuals. Triploid offspring derived from diploid maternal parents had lower germination rates, but higher pollen production than those with tetraploid mothers. Relative to diploids, the cumulative fitness of 2x x 4x triploids and 4x x 2x triploids was 0.12 and 0.06, respectively, providing some support for effect of differing maternal:paternal ratios and endosperm development as a mechanism of hybrid inviability. Collectively, the data show that tetraploids exhibit an inherent fitness disadvantage, although the partial viability and fertility of triploids may help to reduce the barrier to tetraploid establishment in sympatric populations. PMID- 11005288 TI - The evolution of reproductive isolation in the ectomycorrhizal Hebeloma crustuliniforme aggregate (Basidiomycetes) in northwestern Europe: a phylogenetic approach. AB - To reconstruct the evolution of reproductive isolation in the ectomycorrhizal Hebeloma crustuliniforme aggregate (Basidiomycetes), phylogenetic relationships were determined between strains that belong to a clade consisting of nine intercompatibility groups (ICGs, biological species). Four of these nine ICGs are partially compatible and belong to the H. crustuliniforme aggregate. Different levels of partial compatibility have been found between these four ICGs. Between ICGs 3 and 4, 15% of the combinations were compatible. One strain was compatible with all isolates of both ICGs 3 and 4 and also with one isolate of ICG 2. Both a nuclear phylogeny, based on ribosomal IGS sequence data, and a mitochondrial phylogeny, based on a group-I intron located in the large subunit ribosomal RNA gene (LrRNA), were reconstructed. The level of incompatibility was compared with the phylogenetic history of individuals belonging to this clade. Different relationships were found between the level of compatibility and the relative age of the most recent common ancestor (MRCA) for different ICGs. On the one hand, the evolution of incompatibility between ICGs 2 and 3/4 is most consistent with the class of "divergence- first" models because a positive correlation was found between the relative age of the MRCA and the level of incompatibility for ICG 2 versus 3/4. On the other hand, the lack of such a correlation for ICGs 3 and 4 shows that (partial) incompatibility between these ICGs has arisen without strong divergence. The ecological (and to a lesser extent geographical) differences found between ICGs 3 and 4 suggest that selection for incompatibility, associated with host tree preference, has been important in the evolution of incompatibility between these two ICGs. The incongruence between the nuclear and mitochondrial trees for ICG 1 could be explained by a hybrid origin of this ICG, with different donors of the mitochondrial and nuclear sequences. PMID- 11005289 TI - Parental effects in Plantago lanceolata L. III. Measuring parental temperature effects in the field. AB - To determine the evolutionary importance of parental environmental effects in natural populations, we must begin to measure the magnitude of these effects in the field. For this reason, we conducted a combined growth chamber-field experiment to measure parental temperature effects in Plantago lanceolata. We grew in the field offspring of controlled crosses of chamber-grown parents subjected to six temperature treatments. Each treatment was characterized by a unique combination of maternal prezygotic (prior to fertilization), paternal prezygotic, and postzygotic (during fertilization and seed set) temperatures. Offspring were followed for three years to measure the effects of treatment on several life-history traits and population growth rate, our estimate of fitness. Parental treatment influenced germination, growth, and reproduction of newborns, but not survival or reproduction of offspring at least one year old. High postzygotic temperature significantly increased germination and leaf area at 17 weeks by approximately 35% and 2%, respectively. Probability of flowering and spike production in the newborn age class showed significant parental genotype x parental treatment interactions. High postzygotic temperature increased offspring fitness by approximately 50%. The strongest contributors to fitness were germination and probability of flowering and spike production of newborns. A comparison of our data with previously collected data for chambergrown offspring shows that the influence of parental environment on offspring phenotype is weaker but still biologically meaningful in the field. The results provide evidence that parental environment influences offspring fitness in natural populations of P. lanceolata and does so by affecting the life-history traits most strongly contributing to fitness. The data suggest that from the perspective of offspring fitness, natural selection favors parents that flower later in the flowering season in the North Carolina Piedmont when it is warmer. Genotypic-specific differences in response of offspring reproductive traits to parental environment suggest that parental environmental effects can influence the rate of evolutionary change in P. lanceolata. PMID- 11005290 TI - Chloroplast evolution in the Pinus montezumae complex: a coalescent approach to hybridization. AB - This study addresses the evolutionary history of the chloroplast genomes of two closely related pine species, Pinus hartwegii Lindl. and P. montezumae Lamb (subsect. Ponderosae) using coalescent theory and some of the statistical tools that have been developed from it during the past two decades. Pinus hartwegii and P. montezumae are closely related species in the P. montezumae complex (subsect. Ponderosae) of Mexico and Central America. Pinus hartwegii is a high elevation species, whereas P. montezumae occurs at lower elevations. The two species occur on many of the same mountains throughout Mexico. A total of 350 individuals of P. hartwegii and P. montezumae were collected from Nevado de Colima (Jalisco), Cerro Potosi (Nuevo Leon), Iztaccihuatl/Popocatepetl (Mexico), and Nevado de Toluca (Mexico). The chloroplast genome of P. hartwegii and P. montezumae was mapped using eight restriction enzymes. Fifty-one different haplotypes were characterized; 38 of 160 restriction sites were polymorphic. Clades of most parsimoniously related chloroplast haplotypes are geographically localized and do not overlap in distribution, and the geographically localized clades of haplotypes include both P. hartwegii and P. montezumae. Some haplotypes in the clades occur in only one of the two species, whereas other haplotypes occur in both species. These data strongly suggest ancient and/or ongoing hybridization between P. hartwegii and P. montezumae and a shared chloroplast genome history within geographic regions of Mexico. PMID- 11005291 TI - The fitness effects of spontaneous mutations in Caenorhabditis elegans. AB - Spontaneous mutation to mildly deleterious alleles has emerged as a potentially unifying component of a variety of observations in evolutionary genetics and molecular evolution. However, the biological significance of hypotheses based on mildly deleterious mutation depends critically on the rate at which new mutations arise and on their average effects. A long-term mutation-accumulation experiment with replicate lines of the nematode Caenorhabditis elegans maintained by single progeny descent indicates that recurrent spontaneous mutation causes approximately 0.1% decline in fitness per generation, which is about an order of magnitude less than that suggested by previous studies with Drosophila. Two rather different approaches, Bateman-Mukai and maximum likelihood, suggest that this observation, along with the observed rate of increase in the variance of fitness among lines, is consistent with a genomic deleterious mutation rate for fitness of approximately 0.03 per generation and with an average homozygous effect of approximately 12%. The distribution of mutational effects for fitness appears to have a relatively low coefficient of variation, being no more extreme than expected for a negative exponential, and for one composite fitness measure (total progeny production) approaches constancy of effects. These results are derived from assays in a benign environment. At stressful temperatures, estimates of the genomic deleterious mutation rate (for genes expressed at such temperatures) is sixfold lower, whereas those for the average homozygous effect is approximately eightfold higher. Our results are reasonably compatible with existing estimates for flies, when one considers the differences between these species in the number of germ-line cell divisions per generation and the magnitude of transposable element activity. PMID- 11005292 TI - Evolution of poecilogony and the biogeography of North American populations of the polychaete Streblospio. AB - Invertebrate interspecific developmental patterns can be highly variable and, taxonomically, are considered only weakly constrained. Intraspecifically, some invertebrate species possess multiple developmental modes-a condition known as poecilogony. Closer examination of most putative poecilogenous species, however, has not supported poecilogony, but rather has uncovered hidden or cryptic species. The polychaete Streblospio benedicti is a well-known, poecilogenous species found along the coast of North America. We collected mitochondrial cytochrome subunit I DNA sequence data from 88 individuals taken from 11 locations along the Atlantic, Gulf, and Pacific Coasts of the United States to provide a phylogenetic framework from which to interpret intraspecific variation in larval life history and brooding structure morphology in this species. Our results are consistent with a recent revision of the species into two separate species: S. benedicti, a pouched brooding form distributed along the Atlantic and Pacific Coasts, and S. gynobranchiata, a branchiate brooding form in the Gulf of Mexico. Contrary to the redescription, S. benedicti is paraphyletic because the pouched brooding population in Vero Beach, Florida shows strong genetic affinity with Gulf of Mexico populations (S. gynobranchiata). However, S. benedicti is a true poecilogenous species, with both lecithotrophic and planktotrophic individuals possessing identical mitochondrial DNA haplotypes. Crossbreeding experiments further support the molecular phylogeny with reproductive isolation demonstrated between, but not within, the major phylogenetic clades consistent with the previously described species. The genetic break near Vero Beach, Florida, corresponds to a well-known phylogeographic boundary, but the estimated time of separation for the Streblospio spp., approximately 10 million years before present, predates all other known phylogeographic subdivisions in this area. This suggests that biogeographic sundering in this region is a recurrent event. Divergence times within the major Streblospio spp. clades are recent and indicate that changes in larval life history as well as brooding structure morphology are highly plastic and can evolve rapidly. PMID- 11005293 TI - Life-history correlates of evolution under high and low adult mortality. AB - Life-history theory predicts evolutionary changes in reproductive traits and intrinsic mortality rates in response to differences in extrinsic mortality rates. Trade-offs between life- history traits play a pivotal role in these predictions, and such trade-offs are mediated, at least in part, by physiological allocations. To gain insight into these trade-offs, we have been performing a long-term experiment in which we allow fruitflies, Drosophila melanogaster, to evolve in response to high (HAM) and low (LAM) adult mortality rates. Here we analyze the physiological correlates of the life-history trade-offs. In addition to changing development time and early fecundity in the direction predicted, high adult mortality affected three traits expressed early in life-body size, growth rate, and ovariole number-but had little or no effect on body composition (relative fat content), viability, metabolic rate, activity, starvation resistance, or desiccation resistance. Correlations among lines revealed trade offs between early fecundity, late fecundity, and starvation resistance, which appear to be mediated by differential allocation of lipids. PMID- 11005294 TI - Morphological intergration between development compartments in the Drosophila wing. AB - Developmental integration is the covariation among morphological structures due to connections between the developmental processes that built them. Here we use the methods of geometric morphometrics to study integration in the wing of Drosophila melanogaster. In particular, we focus on the hypothesis that the anterior and posterior wing compartments are separate developmental units that vary independently. We measured both variation among genetically diverse individuals and random differences between body sides of single individuals (fluctuating asymmetry, FA). For both of these sources of variation, the patterns of variation identified by principal component analyses all involved landmarks in both the anterior and posterior compartments simultaneously. Analyses focusing exclusively on the covariation between the anterior and posterior compartments, by the partial least-squares method, revealed pervasive integration of the two compartments, for both individual variation and FA. These analyses clearly indicate that the anterior and posterior compartments are not separate units of variation, but that the covariation between compartments is sufficient to account for nearly all the variation throughout the entire wing. We conclude that variation among individuals as well as the developmental perturbations responsible for FA generate shape variation primarily through developmental processes that are integrated across both compartments. In contrast, much less of the shape variation in our sample can be attributed to the localized processes that establish the identity of particular wing veins. PMID- 11005295 TI - Courtship song recognition in the Drosophila melanogaster complex: heterospecific songs make females receptive in D. melanogaster, but not in D. sechellia. AB - The courtship song emitted by male wing vibration has been regarded as one of the most important signals in sexual isolation in the species of the Drosophila melanogaster complex. Inter- and intraspecific crosses were observed using males whose wings were removed (mute) or females whose aristae were removed (deaf). Females of D. melanogaster, D. simulans, and D. mauritiana mated with heterospecific males in the song-present condition (cross between normal females and winged males) more often than in the no-song condition (cross between normal females and wingless males or between aristaless females and winged males) or they showed no preference between the two conditions. It is possible that in these females heterospecific courtship songs play a role as if they were conspecific. In contrast, the females of D. sechellia mated with D. melanogaster or D. simulans males in the no-song condition more often than in the song-present condition, suggesting that they reject males with heterospecific song. Female mate recognition depending on the courtship song in D. melanogaster, D. simulans, and D. mauritiana is considered to be relatively broader and that in D. sechellia narrower. PMID- 11005296 TI - The role of the use of different host plants in the maintenance of the inversion polymorphism in the cactophilic Drosophila buzzatii. AB - Inversion polymorphisms often have been associated with fitness variation. Cactophilic Drosophila buzzatii has been used widely for the study of the maintenance of chromosomal variation. The purpose of this paper is to address the relative importance of variable selection regimes associated with the use of three different host cacti and antagonistic pleiotropy in the maintenance of chromosomal variation. Using homokaryotypic stocks derived from several lines homozygous for four second-chromosome arrangements, we show that inversions significantly affect first-instar larva to adult viability (VT), developmental time (DT) and adult thorax length (TL). We also show that the effects of inversions on DT and VT are dependent on the cactus rearing media. The effects of polymorphic gene arrangements on life-history traits suggest the existence of trade-offs between early and late fitness components. The dosage of arrangement 2st, the ancestral gene order, was negatively correlated with DT and TL, whereas flies carrying the derived arrangements 2j and 2jq7 had longer DTs and larger TLs. Arrangements 2st and 2jq7 increased viability, at least in one of the cactus media tested. Our results suggest that environmental heterogeneity, as represented by the use of different cactus hosts and the trade-off between DT and TL, may be involved in the maintenance of the polymorphism. In addition, our data suggest that the chromosomal phylogeny may be decoupled from the evolution of the genes affecting life-history traits linked to the inversion system. PMID- 11005297 TI - Interspecific genetics of mate recognition: inheritance of female acoustic preference in Hawaiian crickets. AB - Female mating behavior plays a fundamental role in the divergent evolution of mate recognition systems that may lead to speciation. Despite this important role, the phenotypic and genetic bases of female mating behavior remain poorly understood. In this study, I examine the shape of the female acoustic preference function and estimate values for pulse rate preference in two species of Hawaiian crickets, Laupala kohalensis and L. paranigra. In addition, I examine how preference differences are inherited in hybrid crosses between these species. Females expressed unimodal preference functions and were generally more attracted to pulse rates characterizing their own species. Unimodal preference functions also characterized F1 and backcross generations, with hybrid females expressing preferences for intermediate pulse rates. Pulse rate preferences segregated in the backcross generation. Mean pulse rate preference matched mean pulse rate in both parental and hybrid generations. Based on F1 hybrids and segregation patterns in backcross females, I show that changes in both signal and receiver components of the mate recognition system are consistent with a multilocus model of change through incremental steps. The results therefore suggest that ancestors of the current species also expressed unimodal preference functions and that changes in acoustic communication signals occurred through shifts in mean pulse rates and pulse rate preferences among populations. PMID- 11005298 TI - Reproductive character displacement and speciation in periodical cicadas, with description of new species, 13-year Magicicada neotredecem. AB - Acoustic mate-attracting signals of related sympatric, synchronic species are always distinguishable, but those of related allopatric species sometimes are not, thus suggesting that such signals may evolve to "reinforce" premating species isolation when similar species become sympatric. This hypothesis predicts divergences restricted to regions of sympatry in partially overlapping species, but such "reproductive character displacement" has rarely been confirmed. We report such a case in the acoustic signals of a previously unrecognized 13-year periodical cicada species, Magicicada neotredecim, described here as a new species (see Appendix). Where M. neotredecim overlaps M. tredecim in the central United States, the dominant male call pitch (frequency) of M. neotredecim increases from approximately 1.4 kHz to 1.7 kHz, whereas that of M. tredecim remains comparatively stable. The average preferences of female M. neotredecim for call pitch show a similar geographic pattern, changing with the call pitch of conspecific males. Magicicada neotredecim differs from 13-year M. tredecim in abdomen coloration, mitochondrial DNA, and call pitch, but is not consistently distinguishable from 17-year M. septendecim; thus, like other Magicicada species, M. neotredecim appears most closely related to a geographically adjacent counterpart with the alternative life cycle. Speciation in Magicicada may be facilitated by life-cycle changes that create temporal isolation, and reinforcement could play a role by fostering divergence in premating signals prior to speciation. We present two theories of Magicicada speciation by life cycle evolution: "nurse-brood facilitation" and "life-cycle canalization." PMID- 11005299 TI - Genetic evidence for assortative mating between 13-year cicadas and sympatric "17 year cicadas with 13-year life cycles" provides support for allochronic speciation. AB - Thirteen-year cicadas of brood XIX from northern Arkansas, Missouri, and southern Illinois (lineage A) are known to be genetically different at two marker loci (mitochondrial DNA and abdominal color) from 13-year cicadas to the south (lineage B) that emerge in the same year. Because 17-year cicadas from all broods (year classes) are indistinguishable from lineage A at these two marker loci, previous workers suggested that the lineage A cicadas of 13-year brood XIX were derived from 17-year cicadas by life-cycle switching (allochrony). Data presented here show that, over the same northern geographic range, lineage A is also present in 13-year cicadas belonging to brood XXIII (which always emerges four years later than brood XIX). Detailed sampling along the putative life-cycle switching boundary in 13-year brood XXIII revealed a previously unsuspected broad zone of overlap where populations contained individuals of both lineages A and B. Despite this sympatry, and previous reports of a lack of behavioral barriers to interbreeding, a strong correlation between mitochondrial haplotype and abdominal color suggests that assortative mating has taken place. Lineage A 13-year cicadas from both broods XIX and XXIII are only found within a gap in the spatial distribution of 17-year cicadas. This, in combination with the lack of differentiation between lineage A 13- and 17-year cicadas at the marker loci and new behavioral data for 13-year brood XIX, suggests a recent derivation of all northern 13-year cicadas from the 17-year cicadas via life-cycle switching. We discuss the implications of these allochronic shifts for speciation. PMID- 11005300 TI - Tests of pleistocene speciation in montane grasshoppers (genus Melanoplus) from the sky islands of western North America. AB - There has a been a resurgence of debate on whether the Pleistocene glaciations inhibited speciation. This study tests a model of Pleistocene speciation, estimating the phylogenetic relationships and divergence times of 10 species of montane grasshoppers, genus Melanoplus, using 1300 bp of the mitochondrial gene cytochrome oxidase I (COI). Based on average pairwise distances (corrected for multiple substitutions using Kimura's two-parameter model), all species appear to have originated within the Pleistocene. Sequence divergences between species are less than 4%, corresponding to divergence times less than 1.7 million years ago. Branching patterns among the species suggest that speciation was associated with more than one glacial-interglacial cycle. A likelihood-ratio test rejected a model of simultaneous species origins, the predicted branching pattern if species arose from the fragmentation of a widespread ancestor. These grasshoppers live in an area that was previously glaciated and, as inhabitants of the northern Rocky Mountain sky islands, underwent latitudinal and probably altitudinal shifts in distribution in response to climatic fluctuations. Given the repeated distributional shifts and range overlap of the taxa, there most likely has been ample opportunity for population mixing. However, despite periodic glacial cycles, with more than 10 major glaciations over the past million years and climatic fluctuations over as short a time scale as 10(3) to 10(4) years, the dynamic history of the Pleistocene did not preclude speciation. Although relationships among some taxa remain unresolved, these grasshopper species, even with their recent origins, exhibit genetic coherence and monophyletic or paraphyletic gene trees. The frequency of glacial cycles suggests that the speciation process must have been extremely rapid. These species of grasshoppers are morphologically very similar, differing primarily in the shape of the male genitalia. These characters are posited to be under sexual selection, may play an important role in reproductive isolation, and are known to diverge rapidly. This suggests the rapidity of evolution of reproductive isolation may determine whether species divergences occurred during the Pleistocene glaciations. PMID- 11005301 TI - Reinterpreting recapitulation: systematics of needlefishes and their allies (Teleostei: Beloniformes). AB - As needlefishes (Belonidae) grow, their jaws pass through a "halfbeak" stage that resembles the adult jaw condition of the closely related family of halfbeaks (Hemiramphidae). Based on this pattern, some authors have suggested that halfbeaks are "developmentally arrested" or paedomorphic needlefish derivatives, whereas others have supported the notion that needlefishes are descended from halfbeak-like ancestors and that needlefish ontogeny thereby recapitulates phylogeny. To test these ideas and to better understand evolutionary changes in jaw ontogeny, phylogenetic relationships among genera of needlefishes, sauries (Scomberesocidae), halfbeaks, and flyingfishes (Exocoetidae) were assessed using mitochondrial (cytochrome b and 16S), nuclear (Tmo-4C4), and morphological characters. The resultant tree provides several novel taxonomic findings: (1) flyingfishes appear to be nested within halfbeaks; (2) sauries appear to be nested within needlefishes; and (3) the Indo-West Pacific freshwater halfbeaks appear to be most closely related to the needlefish/saury clade. The structure of the tree falsifies the idea that halfbeaks are paedomorphic needlefishes. Instead, halfbeaks are basal relative to needlefishes, fitting the pattern predicted by the hypothesis of recapitulation. I discuss limitations to phylogenetic perspectives on recapitulation based on discrete character data by comparing aspects of von Baerian and Haeckelian views of the relation between ontogeny and phylogeny. PMID- 11005302 TI - Spatiotemporal reorganization of growth rates in the evolution of ontogeny. AB - Heterochrony, evolutionary changes in rate or timing of development producing parallelism between ontogeny and phylogeny, is viewed as the most common type of evolutionary change in development. Alternative hypotheses such as heterotopy, evolutionary change in the spatial patterning of development, are rarely entertained. We examine the evidence for heterochrony and heterotopy in the evolution of body shape in two clades of piranhas. One of these is the sole case of heterochrony previously reported in the group; the others were previously interpreted as cases of heterotopy. To compare ontogenies of shape, we computed ontogenetic trajectories of shape by multivariate regression of geometric shape variables (i.e., partial warp scores and shape coordinates) on centroid size. Rates of development relative to developmental age and angles between the trajectories were compared statistically. We found a significant difference in developmental rate between species of Serrasalmus, suggesting that heterochrony is a partial explanation for the evolution of body shape, but we also found a significant difference between their ontogenetic transformations; the direction of the difference between them suggests that heterotopy also plays a role in this group. In Pygocentrus we found no difference in developmental rate among species, but we did find a difference in the ontogenies, suggesting that heterotopy, but not heterochrony, is the developmental basis for shape diversification in this group. The prevalence of heterotopy as a source of evolutionary novelty remains largely unexplored and will not become clear until the search for developmental explanations looks beyond heterochrony. PMID- 11005303 TI - Evolution of temporal isolation in the wild: genetic divergence in timing of migration and breeding by introduced chinook salmon populations. AB - The timing of migration and breeding are key life-history traits; they are not only adaptations of populations to their environments, but can serve to increase reproductive isolation, facilitating further divergence among populations. As part of a study of divergence of chinook salmon, Oncorhynchus tshawytscha, populations, established in New Zealand from a common source in the early 1900s, we tested the hypotheses that the timing of migration and breeding are under genetic control and that the populations genetically differ in these traits despite phenotypic overlap in timing in the wild. Representatives of families from two populations were collected within a day or two of each other, reared in a common environment, and then released to sea from each of two different rivers, while other family representatives were retained in fresh water to maturity. The date of maturation of fish held in fresh water and the dates of return from the ocean and maturation of fish released to sea all showed significant differences between the two populations and among families within populations. The very high heritabilities and genetic correlations estimated for migration and maturation date indicated that these traits would respond rapidly to selection. Combined with the results of related studies on these chinook salmon populations, it appears that spawning time may not only evolve during the initial phases of divergence, but it may play an important role in accelerating divergence in other traits. PMID- 11005304 TI - Reproductive burden, locomotor performance, and the cost of reproduction in free ranging lizards. AB - A reduction in the locomotor capacity of gravid females is considered to be a cost of reproduction if it leads to an increased risk of mortality. In this study, we measured the change in endurance between gravid and postgravid female side-blotched lizards (Uta stansburiana) as a test of the cost of reproduction. We also altered reproductive investment in some females by direct ovarian manipulation (yolkectomy), which decreased reproductive burden by 30%. Regardless of experimental treatment, all females had lower endurance when gravid. Endurance was 28% lower in gravid females from the yolkectomy treatment and 31% lower in the unmanipulated females relative to postoviposition females. The experimental reduction in clutch mass resulted in a 21% increase in endurance of gravid yolkectomy females relative to control females. Postovipositional endurance was significantly higher in the yolkectomized females than unmanipulated females, which suggests that the cost of reproduction carries over to postoviposition performance. Unmanipulated females exhibited a significant negative association between endurance and size-specific burden. Endurance was not correlated with clutch size or size-specific burden in the yolkectomy females. Survivorship to the second clutch was higher in the yolkectomy females. The results from a logistic regression showed the probability of survival to the second clutch was significantly and positively associated with endurance after controlling for the effects of treatment. Our analyses demonstrated that the decrement in performance associated with current reproductive investment represents a cost of reproduction expressed as diminished locomotor performance and lowered survivorship to the next clutch. PMID- 11005305 TI - Experimental excursions on adaptive landscapes: density-dependent selection on egg size. AB - Theories of density-dependent natural selection suggest that intraspecific competition will favor juveniles of high competitive ability. Empirical evidence has been provided from laboratory selection experiments, but field studies are lacking due to the logistical difficulties of experimentally manipulating population densities in natural settings. Here, we present data from a decade long experimental field study of side-blotched lizards, Uta stansburiana that overcomes these difficulties. We tested the hypothesis that density-dependent natural selection causes egg size to increase from early to late clutches in this and many other species. Using a novel combination of environmental manipulations of hatchling density and phenotypic manipulations of egg size, we demonstrate that the nature of selection on egg size changes dramatically in the absence of older competitors. The strength of selection on egg size among later-clutch hatchlings released in areas without competitors from early clutches became almost doubled in magnitude, compared to that among hatchlings released in the presence of older competitors. These experimental findings demonstrate density dependent natural selection on egg size; however, they contradict the classical idea that egg size increases during the reproductive season because of competition between early and late hatchlings. The results indicate that competitive age or size asymmetries between early and late hatchlings can override within-cohort asymmetries due to egg size. We suggest that competition could be an important mediator of oscillating selection pressures in this and other systems. Finally, we discuss the utility of "double-level," simultaneous experimental manipulation of both phenotypic traits that are targets of selection (e.g., egg size) as well the environmental agents of selection (e.g., population density). PMID- 11005306 TI - Sexual size dimorphism in shorebirds, gulls, and alcids: the influence of sexual and natural selection. AB - Charadrii (shorebirds, gulls, and alcids) have an unusual diversity in their sexual size dimorphism, ranging from monomorphism to either male-biased or female biased dimorphism. We use comparative analyses to investigate whether this variation relates to sexual selection through competition for mates or natural selection through different use of resources by males and females. As predicted by sexual selection theory, we found that in taxa with socially polygynous mating systems, males were relatively larger than females compared with less polygynous species. Furthermore, evolution toward socially polyandrous mating systems was correlated with decreases in relative male size. These patterns depend on the kinds of courtship displays performed by males. In taxa with acrobatic flight displays, males are relatively smaller than in taxa in which courtship involves simple flights or displays from the ground. This result remains significant when the relationship with mating system is controlled statistically, thereby explaining the enigma of why males are often smaller than females in socially monogamous species. We did not find evidence that evolutionary changes in sexual dimorphism relate to niche division on the breeding grounds. In particular, biparental species did not have greater dimorphism in bill lengths than uniparental species, contrary to the hypothesis that selection for ecological divergence on the breeding grounds has been important as a general explanation for patterns of bill dimorphism. Taken together, these results strongly suggest that sexual selection has had a major influence on sexual size dimorphism in Charadrii, whereas divergence in the use of feeding resources while breeding was not supported by our analyses. PMID- 11005307 TI - Microsatellites can be misleading: an empirical and simulation study. AB - It has been long recognized that highly polymorphic genetic markers can lead to underestimation of divergence between populations when migration is low. Microsatellite loci, which are characterized by extremely high mutation rates, are particularly likely to be affected. Here, we report genetic differentiation estimates in a contact zone between two chromosome races of the common shrew (Sorex araneus), based on 10 autosomal microsatellites, a newly developed Y chromosome microsatellite, and mitochondrial DNA. These results are compared to previous data on proteins and karyotypes. Estimates of genetic differentiation based on F- and R-statistics are much lower for autosomal microsatellites than for all other genetic markers. We show by simulations that this discrepancy stems mainly from the high mutation rate of microsatellite markers for F-statistics and from deviations from a single-step mutation model for R-statistics. The sex linked genetic markers show that all gene exchange between races is mediated by females. The absence of male-mediated gene flow most likely results from male hybrid sterility. PMID- 11005308 TI - Population genetic structure of two ecologically distinct Amazonian spiny rats: separating history and current ecology. AB - Population history and current demographic and ecological factors determine the amount of genetic variation within and the degree of differentiation among populations. Differences in the life history and ecology of codistributed species may lead to differences in hierarchical population genetic structure. Here, we compare patterns of genetic diversity and structure of two species of spiny rats in the genus Proechimys from the Rio Jurui of western Amazonian Brazil. Based on the ecological and life-history differences between the two species, we make predictions as to how they might differ in patterns of genetic diversity and structure. We use mitochondrial sequence data from the cytochrome b gene to test these predictions. Although both species maintain nearly the same number of mitochondrial haplotypes across the sampled range, they differ in levels of genetic diversity and geographic structure. Patterns of gene flow are also different between the two species with average M-values of nearly three in P. steerei and less than one in P. simonsi. Our initial predictions are largely upheld by the genetic data and where conflicting hypotheses arise, we suggest further studies that may allow us to distinguish among evolutionary scenarios. Separating the effects of history and ongoing demography on patterns of genetic diversity is challenging. Combining genetic analyses with field studies remains essential to disentangling these complex processes. PMID- 11005309 TI - Heritable variation in a family-diagnostic trait. AB - Derived characters that have not changed during the diversification of a clade provide traits that are diagnostic at higher taxonomic levels. The tetradynamous stamen condition (four long and two short stamens) of the Brassicaceae is an example of a diagnostic trait that has not changed during the diversification of this large flowering plant family. We investigated one hypothesis that might explain the long-term stasis of this trait-that tetradynamous stamens have persisted because of an absence of genetic variation underlying the trait. Through a sib-analysis with Raphanus raphanistrum and an artificial selection experiment with Brassica rapa, we demonstrate that significant genetic variation is present for the tetradynamous condition in both species and that the trait is therefore not constrained from evolutionary change by a lack of heritable genetic variation. PMID- 11005310 TI - Habitat-related adaptive properties of plant cuticular lipids. PMID- 11005311 TI - Effect of stressful and nonstressful growth temperatures on variation of sternopleural bristle number in Drosophila melanogaster. AB - The effect of stressful (31 degrees C) and nonstressful (25 degrees C) growth temperatures on quantitative variation and developmental stability (fluctuating asymmetry) of Drosophila melanogaster was examined in a short-term selection experiment on sternopleural bristle number. Realized heritabilities based on 10 generations of selection in an upward direction did not differ between the two temperature regimes, which indicated that additive genetic variation was not affected by a high, stressful temperature. Phenotypic variability and fluctuating asymmetry of sternopleural bristles were significantly higher under stressful conditions when averaged over generations, although most pairwise comparisons in separate generations showed nonsignificant differences between temperatures. PMID- 11005312 TI - Biogeography of the southeastern United States: a comparison of salamander phylogeographic studies. AB - Most phylogeographic studies of species from the southeastern United States have shown a simple east-west division of mtDNA variation. However, a study of the salamander Ambystoma maculatum resulted in a more complex pattern that includes a close affinity between populations from the Central Highlands of Missouri and Arkansas and the Coastal Plain separated by a genetically distinct central group of populations. We test the generality of this observation by surveying mitochondrial DNA (mtDNA) variation in the closely related species A. talpoideum. An Ambystoma-specific intergenic spacer was amplified and sequenced. The 26 resulting haplotypes varied from 380 to 800 base pairs, and alignments, including the outgroup, required 101 insertions/deletions. Sequence divergence among haplotypes ranged from 0.001 to 0.758. Population subdivision was extensive (theta = 0.64). Phylogenetic analysis of A. talpoideum mtDNA sequence reveals a close relationship between the populations from the Central Highlands and the Coastal Plain. This result is similar to that obtained for A. maculatum, although the A. talpoideum clade is not as well differentiated from its sister clades. We discuss the differences and similarities between the two Ambystoma species and previous studies and call for increased focus on multiple species with similar ecologies as a way to detect subtle biogeographic events. PMID- 11005313 TI - Modulation of the T cell receptor beta chain repertoire after heart transplantation. AB - BACKGROUND: In a previous study it was shown that pre-transplant blood transfusion was associated with a better clinical outcome after heart transplantation (HTx). In this study the effect of heart transplantation (HTx) on the T cell receptor V beta chain (TCRVbeta) repertoire was investigated. Therefore, we analyzed the TCRVbeta repertoire of patients after HTx to see whether a correlation with clinical outcome could be observed. METHODS: Patients were analyzed at four different time points: pre-HTx, less than 1 month post-Htx, between 1 month and 2.5 month post-Htx and more than 2.5 months post-HTx. CD4+ and CD8+ T cells were purified from patient peripheral blood mononuclear cells (PBMC). TCR beta chain usage was analyzed semiquantitatively by Southern blot analysis. RESULTS: HTx affected the TCRVbeta repertoire in both the CD4+ and CD8+ T cell compartments in all patients. Changes in the TCRVbeta repertoire were most pronounced within the CD8+ T cell subset. Interestingly, one patient showed modulation in TCRVbeta chain usage predominantly in the CD4+ T cell compartment. CONCLUSIONS: Modulation of TCRVbeta chain usage was detected in all patients analyzed. No clear-cut relation was observed between TCRVbeta modulation after transplantation and clinical outcome. In some cases modulations appeared to concur with observed immunological events (clinically and/or in-vitro). PMID- 11005314 TI - Clusterin may be involved in rat liver allograft tolerance. AB - Little is known about the possible role of complement inhibitors on tolerance induced by liver allografts. Clusterin, which is a plasma glycoprotein, inhibits cytolytic membrane attack complex (MAC) of complement by binding to soluble C5b-7 complex. The role of clusterin in relation to the naturally achieved tolerance in a rat orthotopic liver transplantation (OLT) has not been investigated before. Here we determined the kinetics of clusterin expression at different post transplantation time points in a tolerogenic model (DA-PVG) where rejection was naturally overcome without any immunosuppressive drugs in comparison with the syngenic OLT model (DA-DA). Peripheral blood and liver tissues were taken from OLT at various post-operative time points. A strong expression of soluble clusterin was observed on post-transplantation day 7, which occurred at the peak of the rejection in this tolerogenic OLT model. The expression of clusterin remained strong even after tolerance was achieved. The intensity of clusterin expression was much stronger when compared with the syngenic OLT (DA-DA) model after OLT. A strong expression of clusterin mRNA was also observed in the tolerogenic model on post-OLT day (POD) 7 and the expression persisted when compared with the syngenic model on post-OLT day 60. Our data have shown that the strongest levels of clusterin during the reaction phase in tolerogenic OLT may be involved in tolerance induction. PMID- 11005315 TI - Anti-CD4 induced rat heart tolerance: no presence of primed T cells and regulatory mechanisms for cytotoxic T cells. AB - Treatment with anti-CD4 monoclonal antibody (mAb) (OX38) induces heart, but not skin graft tolerance in WF (RT1u) to Lewis (RT1l) rat strain combinations. We examined differences in cellular responses between heart-bearing and skin rejected hosts that were both treated with anti-CD4 mAb. In the tolerant LEW rats bearing WF heart transplants, the secondary WF heart but not skin grafts were accepted. On the other hand, in anti-CD4 treated WF skin-rejected hosts, both secondary WF heart and skin grafts were rapidly rejected. Spleen cells from anti CD4 treated WF skin-rejected LEW rats but not from WF heart-bearing LEW rats received the same treatment generated CTL after in vitro stimulation with paraformaldehyde (PFA) treated donor WF stimulator spleen cells. Adoptive transfer of spleen cells from WF skin-rejected LEW rats with or without anti-CD4 therapy into the tolerant LEW rats at the secondary WF heart transplantation blocked the secondary heart graft acceptance. However, transfer of spleen cells from WF heart-rejected rats without immunosuppression failed to block acceptance of the secondary heart graft. Our results indicated the lack of primed T cells and presence of regulatory mechanisms for tissue specific T cells in anti-CD4 treated heart bearing hosts. PMID- 11005316 TI - Cytokine secretion in mixed lymphocyte culture: a prognostic indicator of renal allograft rejection in addition to HLA mismatching. AB - We have previously demonstrated significant inter-individual variations in cytokine protein secretion between normal individuals and patients prior to renal transplantation. In this study, pre-transplant patient vs. donor mixed lymphocyte cultures (MLC) were set up between 57 renal allograft patient/donor pairs, and secretion of cytokine protein (IL-2, IL-4, IL-6, IL-10 and IFN-gamma) into the culture supernatant measured by ELISA. Significant inter-individual variations in protein secretion in MLC were observed for all cytokines studied. Univariate analysis demonstrated that high levels of IFN-gamma and IL-10 in MLC and spontaneous IL-4, together with female donor sex and a high degree of HLA mismatching (especially HLA-DR) were significantly associated with rejection. However, multivariate analysis revealed the greatest risk of rejection (RR = 25.5, P = 0.003) was associated with a combination of high IL-10 secretion in MLC and mismatching for at least four HLA antigens (HLA-A, -B and -DR). It remains to be determined whether cytokine secretion in MLC is linked to cytokine gene polymorphisms. In future, assays for measuring either cytokine secretion or genetic polymorphisms may prove to be useful in aiding donor selection and tailoring immunosuppressive therapy. PMID- 11005318 TI - Flow-PRA evaluation for antibody screening in patients awaiting kidney transplantation. AB - Flow-PRA is a flow cytometric method for both anti-HLA class I and class II antibody (Ab) detection. We evaluated this technique for Ab screening in patients awaiting kidney transplantation. After having established a rigorous threshold for positivity, a three-dilution difference in sensitivity between Flow-PRA and complement-dependent cytotoxicity (CDC) persisted. The sensitivity of the method was satisfactory since all CDC-positive sera were also found to be positive with the Flow-PRA method. Discrimination between anti-HLA class I and class II Abs was excellent. Furthermore, all sera responsible for a positive flow cytometry crossmatch (FCXM) and a negative CDC-crossmatch (CDCXM) at the time of a putative transplant were found to be positive with Flow-PRA beads. The specificity was excellent for anti-class I Ab detection since no false positive serum was found. On the other hand, the specificity was lower for anti-class II detection, since 8.3% (2/24) false positive results were detected among all the negative sera tested. Overall, our results suggested that Flow-PRA should be of value for anti HLA Ab screening prior to kidney transplantation. PMID- 11005317 TI - The peripheral lymphocyte count predicts graft survival in DA to Lewis heterotopic heart transplantation treated with FTY720 and SDZ RAD. AB - OBJECTIVE: The new immunomodulator 2-amino-2-(2-[4-octylphenyl]ethyl)-1,3 propanediol hydrochloride (FTY720) lowers the peripheral lymphocyte count (PLC) by inducing migration of circulating lymphocytes to secondary lymphoid organs. This effect is dose-dependent at low (up to 0.1 mg/kg per day) doses in rats. We investigated the correlation between PLC and the later rejection, when FTY720 was combined with RAD. METHODS: Heterotopic cardiac grafting was performed using the DA-Lewis strain combination. FTY720 and RAD were administered as single daily doses by gavage alone and in combination starting 3 days before to 28 days after transplantation. Graft survival was monitored daily by palpation. PLC was determined at 1 and 4 weeks, body weight (BW) weekly. Histologic evaluation of grafted hearts was performed after rejection. MAIN FINDINGS: FTY720 at doses of 0.03, 0.1 and 0.3 mg/kg per day prolonged graft survival dose-dependently from 6 (placebo) to 7, 9.5 and 15 days median survival time (MST). RAD at doses of 0.3, 1 and 3 mg/kg per day delayed rejection to 8.5, 18 and 37.5 days MST. Very small FTY720 doses added to the lower RAD doses were effective in maintaining grafts throughout the treatment period and with normal weight gain, as opposed to regimens with 1 mg/kg or more per day RAD, which resulted in delayed weight gain. FTY720 lowered the PLC significantly and dose-dependently. The PLC correlated well with graft survival [Spearman rank correlation (n = 30, rs = -0.75)]. CONCLUSIONS: Fully effective FTY720 + RAD combination regimens caused no side effects with respect to the rats' general well-being or weight gain and were better tolerated than equiactive RAD monotherapy, suggesting a broader therapeutic window for the combinations. Under the experimental conditions, the PLC decrease showed an interesting correlation with the anti-rejection effects in these two-drug regimens. Thus, in rats the PLC is helpful for monitoring the biological activity of FTY720 at low doses (< 0.1 mg/kg per day), i.e. in the range of the steep part of its dose-response relationship. PMID- 11005320 TI - Genetic conservation of the immunophilin-binding domains of human calcineurin A1 and A2. AB - Calcineurin a calmodulin-dependent phosphatase plays a critical role in calcium dependent activation of T-lymphocytes and is the major target for the inhibitory actions of the immunosuppressive drugs Tacrolimus (FK506) and Cyclosporin A (CsA). Calcineurin is a dimeric protein consisting of distinct A (catalytic) and B (regulatory) subunits. In humans two separate genes, CNA1 and CNA2, encode the calcineurin A (CNA) subunit. The region of CNA that interacts with Calcineurin B, calmodulin, and immunosuppressive drugs bound to their receptors--the immunophilins--has been identified to amino acids 281-414 (Greengard P, Allen PB, Nairin AC. Beyond the dopamine receptor: the DARPP-32/protein phosphatase-1 cascade. Neuron 1999;23:435). Our working hypothesis was that the differences in patient response to calcineurin inhibitors could be a consequence of inherited variations within their CNA genes. Single-strand conformational polymorphism (SSCP) analysis of cDNAs derived from the coding region for amino acids 281-414 of CNA1 and CNA2 in 32 healthy Caucasians did not detect polymorphic variations within these genes. These results suggest that this region is highly conserved and cannot account for individual variation in response of patients to FK506 and CsA treatment. PMID- 11005319 TI - Anti-galactose-alpha(1,3) galactose antibody production in alpha1,3 galactosyltransferase gene knockout mice after xeno and allo transplantation. AB - Antibodies (Abs) that mediate the hyperacute rejection and acute vascular rejection/delayed xenograft rejection of pig organs in humans and Old World primates are predominantly directed at a single carbohydrate epitope, galactose alpha1,3-galactose (alpha1,3Gal). The T-cell dependence of elicited anti alpha1,3Gal Ab responses in humans and Old World primates is controversial. In this study we have characterized anti-alpha1,3Gal Ab production in mice with disrupted alpha1,3-galactosyltransferase genes (GT-Ko mice) and determined the T cell dependence of anti-alpha1,3Gal Ab responses, following xenograft and allograft transplantation. GT-Ko mice produce natural anti-alpha1,3Gal IgM and IgG in an age-dependent manner, however, these Abs could not elicit hyperacute rejection nor affect the rate of cardiac xenograft (3-5 days) or allograft rejection (7-9 days). Transplantation of xenogeneic Lewis rats hearts elicited modest anti-alpha1,3Gal Ab, but vigorous xenoAb responses. The anti-alpha1,3Gal Ab response was restricted to the IgM and IgG3 subclass while the xenoAb response comprised IgM and all four IgG subclasses. Transplantation of allogeneic C3H hearts elicited weak anti-alpha1,3Gal Ab responses that were primarily IgM, but vigorous alloAb responses. Despite the restriction of elicited anti-alpha 1,3Gal Ab responses to the IgM and IgG3 isotypes, these responses are T-cell dependent. The ability of allografts to elicit weak anti-alpha1,3Gal but strong allo-Ab responses, can be explained by the dependence of alpha1,3Gal-specific B cells on cognate help from T cells. PMID- 11005321 TI - Levels of mycophenolic acid and its glucuronide derivative in the plasma of liver, small bowel and kidney transplant patients receiving tacrolimus and cellcept combination therapy. AB - In order to help assess the usefulness of mycophenolate mofetil (MMF) as an immunosuppressive agent in recipients of organs other than kidneys, we measured the trough levels of the active metabolite of MMF, mycophenolic acid (MPA), and its inactive glucuronide derivative (MPAG), in the plasma of liver (n = 83) and small bowel transplant patients (n = 15) receiving MMF in combination with tacrolimus. These levels were compared with a group of renal transplant patients (n = 25) receiving the same drug regimen. All patient groups were otherwise comparable except the small bowel patient group which contained more pediatric patients (average age 18.7 +/- 3.9 years), and, therefore, received a higher average drug dose (in mg/kg). Despite this, these patients displayed the lowest levels of MPA of any group (0.39 +/- 0.08 microg/ml, P < 0.001 vs. 1.10 +/- 0.17 microg/ml for liver transplant patients, P < 0.001 or 2.46 +/- 0.37 microg/ml for renal transplant patients, P < 0.001). There were no statistically significant differences in MPAG levels between any of the groups. Although preliminary, these data demonstrate significant transplanted organ-specific differences in MMF pharmacology and/or bioavailability, and suggest the need for separate evaluation of MMF dosing for each transplant type. PMID- 11005322 TI - Cyclosporine A sensitivity in vitro and P-glycoprotein expression in patients on dialysis and after kidney transplantation. AB - BACKGROUND: In allogeneic kidney transplantation the response to cyclosporine A (CsA) is important for graft outcome. Although CsA therapy is controlled by drug monitoring to ensure therapeutic CsA levels, the sensitivity to the effects of CsA varies among individuals. Since CsA is an antagonist of cytostatic drugs in P glycoprotein (Pgp)-mediated transport, increased Pgp expression might contribute to an increased resistance to CsA. METHODS: The sensitivity of lymphocytes at three different concentrations of CsA was tested in a non-radioactive lymphocyte transformation test and related to Pgp expression as determined by flow cytometry on mononuclear cells. Five groups, including healthy donors (CON; n = 25), patients on dialysis (DIAL; n = 25), patients before transplantation (PTX; n = 5) and after transplantation [short-term (ATX; n = 5) and long-term (LTX; n = 25)] were investigated. RESULTS: In LTX, the sensitivity to CsA at 400 and 1000 ng/ml was significantly different from CON and DIAL. Overall a higher sensitivity to CsA was seen in patients after transplantation. In ATX, sensitivity to CsA was significantly higher than in PTX at a concentration of 1000 ng/ml CsA. However, comparing all groups no significant changes in Pgp expression were noted. Analysing the relationship between CsA sensitivity and Pgp expression, no significant heterogeneity could be observed between the different groups. CONCLUSION: In conclusion, our data suggest that in vitro testing of CsA sensitivity prior transplantation and Pgp expression monitoring yield independent results and cannot substitute for each other as predictors of graft outcome. The differential role of each test for the evaluation of CsA sensitivity or resistance remains to be determined. PMID- 11005323 TI - Treatment of chronic contained spontaneous esophageal perforations. AB - Spontaneous esophageal perforations are associated with a high mortality and morbidity without surgery. The treatment mortality for early (<24) and late (>24 h) spontaneous esophageal perforations is reviewed as well as all recent cases of chronic spontaneous esophageal perforations. Chronic esophageal perforations with mediastinal cavities may be best treated by internal drainage of the cavity into the esophagus in order to convert the transmural perforation into an intramural esophageal dissection. PMID- 11005324 TI - Prevalence of Barrett's esophagus by endoscopy and histologic studies: a prospective evaluation of 306 control subjects and 376 patients with symptoms of gastroesophageal reflux. AB - The classic endoscopic diagnosis of a Barrett's esophagus (BE) is based on the finding of > or =3 cm, of distal esophagus covered by specialized columnar epithelium. However, currently, it is based on the finding of intestinal metaplasia (IM) at the squamous-columnar mucosal junction, independent of its extent. The aim of this study was to determine the prevalence of Barrett's esophagus by endoscopic and histological findings in control subjects and in patients with symptoms of gastroesophageal reflux (GER). Three hundred and six control subjects and 376 patients with symptoms of gastroesophageal reflux were included in this prospective study. Patients with Barrett's esophagus were classified in three groups as follows. 1. Intestinal metaplasia at the cardia. When endoscopy showed non-Barrett's esophagus, but histological intestinal metaplasia was found. 2. Short-segment Barrett's esophagus. When <3 cm, was covered with tongues or finger-like or creeping substitution of distal esophagus. 3. Long-segment Barrett's esophagus. When > 3 cm, of distal esophagus was covered by specialized columnar epithelium. Two biopsies at the antrum, four biopsies at the squamous-columnar junction and one or two at the distal esophagus were taken. In control subjects, 1.6% showed histological IM at the esophagogastric junction. In patients with GER without esophagitis or with erosive esophagitis, IM was found in 18% and 10.7% respectively. 'Short-segment' Barrett's esophagus was three times more frequent than 'long-segment' Barrett's esophagus. Patients with Barrett's esophagus were significantly older than the other groups. The presence of complications or erosions, peptic ulcer or stricture were significantly more frequent among patients with 'long-segment' Barrett's esophagus (p < 0.0001). The prevalence of dysplasia was similar in all groups of patients with Barrett's esophagus. Complications such as ulcers, stricture and dysplasia were exclusively seen among patients with BE, whereas non-Barrett's patients did not exhibit these complications. In control subjects, IM can be found in a low percentage of cases. Among patients with symptoms of GER, the classic endoscopic diagnosis of a Barrett's esophagus can underestimate this condition in 80% of the cases. Patients with intestinal metaplasia at the cardia already present 17% of the cases with low-grade dysplasia. In all patients with symptoms of GER, systematic biopsies at the squamous-columnar junction should be taken. PMID- 11005325 TI - Antireflux surgery, highly selective vagotomy and duodenal switch procedure: post operative evaluation in patients with complicated and non-complicated Barrett's esophagus. AB - Antireflux surgery, highly selective vagotomy (HSV) and Roux-en-Y duodenojejunostomy have been suggested for control of pathophysiological factors involved in patients with Barrett's esophagus (BE). The aim of this study was to evaluate prospectively the results of this technique in patients with complicated (n = 21) and noncomplicated (n=45) BE. Complete evaluation of esophageal function, endoscopic histologic and clinical control was carried out before and 2 years after surgery. Post-operative results show recurrence of ulcer in patients with complicated BE, but no recurrence in patients with non-complicated BE. Preoperative esophageal ulcer and stricture were present in 85.3% and 14.3%, respectively, of patients with complicated BE. In this group, recurrence of these complications was 38.1% and 9.5% respectively. The technique offers excellent results in patients with non-complicated BE. However, in patients with complicated BE, the recurrence rate is higher, mainly because of the persistence of acid reflux into the esophagus. PMID- 11005326 TI - Aminolevulinic acid-induced photodynamic therapy: safe and effective ablation of dysplasia in Barrett's esophagus. AB - Photodynamic therapy (PDT) is a treatment in which cell damage is achieved by the action of light on a photosensitizer. The aim of this study was to assess the value of PDT in the treatment of Barrett's esophagus with low-grade dysplasia (LGD). Forty patients with dysplastic Barrett's esophagus were given aminolevulinic acid (ALA) 30 mg/kg orally followed 4 h later by laser endoscopy. Follow-up endoscopy and biopsies were performed at 1, 6 and 12 months. A macroscopic response was seen in 33 out of 40 patients, with a median decrease in columnar epithelial area of 30% (range 0-90%). Post-treatment biopsies showed no dysplasia in 39 out of 40 patients, with LGD remaining in one case. This was maintained at 6 and 12 months. There were no significant side-effects. This study demonstrates that ALA-induced PDT provides safe and effective ablation therapy for Barrett's esophagus and is particularly useful against dysplasia. PMID- 11005327 TI - Barrett's mucosa, Barrett's dysplasia and Barrett's carcinoma: diagnostic endoscopy without biopsy-taking does not suffice. AB - Many gastroenterologists are of the opinion that endoscopic diagnosis of gastro oesophageal reflux disease (GORD) suffices and that additional biopsies are not necessary. The data obtained from 1068 consecutive patients with histologically confirmed Barrett's oesophagus were analysed retrospectively. In 37.9% of the patients, the histological diagnosis of Barrett's oesophagus was an incidental finding, whereas 32.7% of Barrett's carcinomas were diagnosed only at histology but not during endoscopy. Of the Patients with dysplasia, 92.4% were diagnosed only by the pathologist. Our analysis shows that an endoscopic diagnosis suspicious for Barrett's mucosa is made in 62.1% of the cases, carcinoma in 70%, and dysplasia in only 7.6% of the cases. Also, because neoplasia is detected for the most part at the invasive carcinoma state, but not in the dysplasia stage, the diagnosis of Barrett's oesophagus, with and without dysplasia, needs to be improved by additional biopsies for histopathological investigation. PMID- 11005328 TI - Incidence of adenocarcinoma and mortality in patients with Barrett's oesophagus diagnosed between 1976 and 1986: implications for endoscopic surveillance. AB - The objective of this study was to investigate the incidence of oesophageal adenocarcinoma and its contribution to mortality in patients with Barrett's oesophagus, and to identify a subgroup of patients who may benefit from endoscopic surveillance. This was a retrospective study of a cohort of 70 patients diagnosed in the endoscopy unit of a Scottish teaching hospital as having Barrett's oesophagus between 1976 and 1986. Information was obtained from case notes, endoscopy records, histology reports and death certificates. Patients were included if they had: (a) columnar-lined oesophagus of at least 3 cm; (b) histological confirmation; and (c) absence of cancer when endoscopically diagnosed as having Barrett's oesophagus. The main outcome for the patients was development of adenocarcinoma or death. Information was available for 59 of 70 patients (84%). Forty-four patients were confirmed to have Barrett's oesophagus and were followed up for 418 patient-years. Only two patients developed oesophageal adenocarcinoma, resulting in an incidence of one case in 209 patient years, a 55-fold risk compared with age- and sex-matched population in Scotland. Both these patients had intestinal metaplasia and Barrett's ulcer. The majority (90%) of patients died as a result of causes unrelated to adenocarcinoma of oesophagus. In patients under 70 years with intestinal metaplasia, 189 annual endoscopies would have been required to detect one cancer. Adenocarcinoma is an uncommon cause of mortality in patients with Barrett's oesophagus. Endoscopic surveillance is unlikely to alter overall mortality in patients with Barrett's oesophagus. However, by limiting endoscopic surveillance to patients under 70 years with intestinal metaplasia, the estimated cost per cancer detected can be reduced to l23000. PMID- 11005329 TI - Recurrence and survival after resection of adenocarcinoma of the gastric cardia. Rotterdam Esophageal Tumor Study Group. AB - In a retrospective study, the results after resection of carcinoma of the gastric cardia in the era without neoadjuvant therapy or extended lymph node dissection were evaluated. All 184 patients who underwent resection between January 1983 and December 1993 were included. Recurrence of disease, survival and prognostic factors were determined. The overall cumulative 5-year recurrence rate was 71% and the survival rate 23%. Multivariate analysis identified locoregional lymph node and distant metastases as the crucial prognosticators of recurrence of disease and survival. These results were similar to those from a previous study concerning our patients operated during the years 1983-88. The prognosis of a resected cardiacarcinoma has remained unchanged in our hands over the past 10 years. These results stress the importance of exploring new ways, such as the use of new diagnostic tools, to optimize preoperative patient selection and more aggressive treatment regimens to improve final outcome. PMID- 11005330 TI - Evaluation of the intestinal microflora in the rat model for esophageal adenocarcinoma. AB - Surgically induced duodenal reflux results in cancer development in the rat esophagus. One proposed mechanism of carcinogenesis relies on the production of carcinogens in the presence of bacterial overgrowth. Against this background, intestinal microflora in the rat jejunum was analyzed before and after reflux inducing surgery. Total gastrectomy and esophagojejunostomy were performed on Sprague-Dawley rats to produce esophageal reflux of duodenal juice (n = 12). Three days before surgery they were randomized into three groups: animals which received tap water; animals which received acidified water at pH 1.8; and animals subjected to oral decontamination with triple antibiotics. During surgery and at autopsy after 2 weeks, intestinal juice was aspirated and analyzed immediately for bacterial content. The physiologic microflora of the rat jejunum contained Lactobacillus spp. and Bacteroides spp., both of which were resistant to the antibiotic regimen. Bacterial overgrowth with fecal bacteria was found following surgery. Acidified water did not alter the intestinal microflora. Triple antibiotics eliminated Escherichia coli and Proteus spp. and reduced the concentration of Enterococcus spp. Bacterial overgrowth by bacteria of the fecal flora occurs in the rat model of esophageal adenocarcinoma with the potential to catalyze the production of carcinogens. PMID- 11005331 TI - Detection of bile reflux: in vivo validation of the Bilitec fibreoptic system. AB - The Bilitec ambulatory bile reflux monitor is advocated for the assessment of bile reflux. However, it has only been validated by bench top (in vitro) studies. This in vivo study used controlled 'iatrogenic' episodes of bile reflux to determine the ability of the Bilitec system to detect bile acids in the oesophageal lumen. At least 10 ml of fluid was aspirated from the stomach of each of 32 patients, and analysed quantitatively for total bile acids (TBA) and total bilirubin (TB). An in vitro assessment of this fluid with the Bilitec probe was performed initially. The Bilitec probe was then sited in the distal oesophagus, and the gastric fluid was introduced into the mid-oesophagus, through a naso oesophageal tube, as a series of 10-ml volumes of varying dilutions (1:8, 1:4, 1:2, undiluted) to simulate gastro-oesophageal reflux events. Intraoesophageal bilirubin absorbance was compared with TBA and TB for each dilution. Both TBA and TB values correlated strongly with bilirubin absorbance in vitro (r = 0.83, p < 0.0001 and r = 0.82, p < 0.0001 respectively). However, this relationship was weaker in vivo (r = 0.64, p = 0.0001 and r = 0.68, p < 0.0001 respectively). Of the dilutions which contained potentially injurious concentrations of bile acid (>1 mmol/L), 77% were associated with absorbance >0.14 (currently accepted absorbance threshold for oesophageal bile reflux). Although in vitro sensitivity of the Bilitec probe to bile acids was excellent, in vivo sensitivity was less reliable, and 23% of significant 'bile reflux' episodes did not exceed the currently recommended Bilitec absorbance threshold for bile reflux. In contrast, false-positive results were unlikely. These findings suggest that the overall accuracy of the Bilitec system is probably sufficient for clinical use, although this method is not sufficiently reliable to be regarded as the gold standard for the evaluation of duodeno-oesophageal reflux. PMID- 11005332 TI - Reflux and pH: 'alkaline' components are not neutralized by gastric pH variations. AB - The ability of the 'alkaline' components of reflux to cause harm in vivo is still open to debate, although these components have been shown in vitro to be capable of damaging the mucosa. The precipitation of bile acids and lysolecithin that occurs at low pH values is the main reason for questioning in vivo mucosal damage. This study was undertaken to determine the composition of gastric aspirates at different original pH values and the degree of solubility of the alkaline components when pH modifications are artificially induced. The samples for chemical analysis were collected from indwelling nasogastric tubes after surgical procedures that did not involve the upper gastrointestinal tract. Bile acid and lysolecithin concentrations were assessed by means of dedicated methods. Thirty-five samples were available for bile acid evaluation and 27 for lysolecithin evaluation. Bile acid and lysolecithin assessments were repeated after pH adjustment at 2, 3.5, 5.5 and 7. For easier assessment of the results, three ranges of the original pH were selected (pH < 2, 2 < or = pH < 5, pH > or = 5). For each pH range, results were pooled together and compared with those in the other pH ranges. Bile acid concentrations were 113+/-48, 339+/-90 and 900+/ 303 (mean +/- s.e.m. micromol/L), respectively, in the three groups selected on account of the different original pH values. Differences were significant (p < 0.001). Both taurine- and glycine-conjugated bile acids were represented even at pH < 2. No major differences were observed in bile acid concentration with the artificially induced pH variations. Lysolecithin concentrations were 5.99+/-3.27, 30.80+/-8.43 and 108.37+/-22.17 (mean +/- SEM microg/ml), respectively, in the three groups selected on account of the different original pH ranges. Differences were significant (p < 0.001). No significant differences in lysolecithin concentration were detected with the artificially induced pH variations. In conclusion, both bile acids and lysolecithin are naturally represented in the gastric environment even at very low pH values, although their concentrations decrease on lowering of the naturally occurring pH. Given the concentration variability of bile acids and lysolecithin, further studies are needed to assess the minimal concentration capable of mucosal damage in vivo. PMID- 11005333 TI - Fundal gastritis as a potential cause of reflux oesophagitis. AB - The transient lower oesophageal sphincter relaxations which allow reflux may be due to altered afferent pathways from the fundus. We aimed to determine whether fundal inflammation is the underlying cause. Two endoscopic biopsies were taken from each of the gastric antrum and fundus in 25 asymptomatic controls with a normal endoscopy (median age 54 range 13-83 years), and 33 patients with erosive oesophagitis (median age 52, 11-78 years). No patient had taken acid suppression therapy or antibiotics for at least 1 month. Sections were stained with haematoxylin and eosin and Giemsa stain and examined in a blinded fashion by one pathologist for the presence of gastritis (Sydney classification) and Helicobacter pylori. Chronic gastritis was common in both groups, but was usually mild. In Helicobacter pylori-negative subjects, there was significantly less chronic gastritis in the antrum and the fundus in oesophagitis patients than in controls (p < 0.05). When present, gastric atrophy was usually antral and mild in severity. There was no difference in the incidence of gastric atrophy in patients with oesophagitis compared with controls (24% compared with 40%; p > 0.05). Chronic gastritis is not more common in patients with oesophagitis, and is unlikely to play a part in the pathogenesis of this disease. PMID- 11005334 TI - Comparison of clinical, endoscopic and functional findings in patients with intestinal metaplasia at the cardia, carditis and short-segment columnar epithelium of the distal esophagus with and without intestinal metaplasia. AB - In recent years, the diagnosis of short segments of intestinal metaplasia lining the distal esophagus has increased. The aim of the present study was to determine the clinical, endoscopic, histologic and functional results in patients with intestinal metaplasia at the cardia (IMC), carditis and short-segment columnar epithelium (CE) lining the distal esophagus with and without intestinal metaplasia. Four groups were studied: 48 patients with carditis, 105 patients with IMC, 78 patients with short-segment CE (SSCE) without IM and 69 patients with short-segment CE with IM. All had clinical questionnaire, endoscopic and histological evaluation, manometric studies and measurements of acid and bilirubin exposition of the distal esophagus over 24 h. Patients without IM were found to be younger than those with IM. Erosive esophagitis was observed in similar proportions, but hiatal hernia was present in patients with SSCE with or without IM. Patients without IM had mainly cardial mucosa more than fundic mucosa. However, patients with IM had almost exclusively cardial mucosa. Low grade dysplasia was observed only in patients with IM. Manometric evaluation demonstrated a structural defective lower esophageal sphincter in all groups. Acid and duodenal exposures of the distal esophagus over 24 h were significantly greater in patients with SSCE with IM. In the presence of pathologic gastroesophageal reflux (GER), there are several histological changes at the mucosa distal to the squamous columnar junction. The first metaplastic change is one from fundic to cardial mucosa and, when duodenal reflux occurs, a second metaplastic change to intestinal metaplasia from cardial mucosa occurs. Therefore, in all patients with symptoms of GER, biopsies specimens distal to the squamous columnar junction should be taken routinely. PMID- 11005335 TI - The effect of cola consumption on oral mucosa in rats. AB - Drinks that contain phosphoric acid have been shown to have erosive effects and cola drinks are strongly acidic (pH 2.5). Gingivitis may be caused by dietary acids. Therefore, this study analyses the interaction of Coca Cola consumption and oral mucosal damage. Thirty rats were divided into three groups of 10. The animals received saline (pH 7.0) or HCl acid buffered to pH 2.6 or Coca Cola (pH 2.6) per os with 24-h free access to these solutions. A biopsy was taken from the front of the gingiva and the tongue. Histopathological analysis showed no specific lesion and there were no differences among saline, Coca Cola and HCl groups. Flow cytometric analysis was used to assess proliferative activity. In the HCl acid and Coca Cola groups, cell cycle analysis showed that the effects of Coca Cola and HCl acid in inducing oral mucosal damage are similar. In both Coca Cola [G0/G1, 70.38+/-7.9; S, 28.06+/-10.13; G2/M, 1.62+/-2.80; proliferative index (PI), 28.68+/-7.981 and HCI (G0/G1, 67.7+/-18.9; S, 27.8+/-17.5; G2/M, 4.4+/-3.8; PI, 30.9+/-20.98), the rat cell population G0/G1 and G2/M phases were found to be low (p < 0.05) and the cell population S and PI phases were found to be significantly elevated compared with the control group (p < 0.05) (G0/G1, 86.92+/-8.69; S, 9.8+/-1.21; G2/M, 3.25+/-2.87; P1, 13.2+/-8.7). This result was reflected in the proliferative index, which is used as a measure of the regeneration index. The data show that Coca Cola and HCl acid have similar proliferative and regenerative effects on oral mucosa, and it is possible that their regenerative effects are caused as a result of an irritant effect. PMID- 11005336 TI - Esophagogastric fistula secondary to teflon pledget: a rare complication following laparoscopic fundoplication. AB - Laparoscopic fundoplication has become the standard operation for gastroesophageal reflux disease. In our service, a laparoscopic fundoplication is performed as a 2-cm floppy 360 degrees wrap with division of the short gastric vessels and the fundoplication is sutured using a prolene 2/0 mattress suture (Ethicon, USA) and buttressed laterally with two teflon pledgets (PTFE 1.85 mm; low porosity, Bard, USA). We report a patient with post-operative dysphagia due to an esophagogastric fistula caused by erosion of a teflon pledget. This is the first such case in 734 laparoscopic fundoplications performed between January 1991 and December 1998. Reoperation was required, resulting in a prolonged convalescence. A review of current literature has not revealed any similar cases. Causes for this rare complication are postulated. PMID- 11005337 TI - Inflammatory fibroid polyp of the esophagus. AB - The case of a 76-year-old woman with a submucosal tumor of the esophagus, whose principal symptoms were dysphagia and epigastric/retrosternal pain, is reported here. Endoscopy, barium swallow and a CAT scan all pointed to extramucosal localization. The lesion was located in the lower esophagus lying on the stomach fundus. An ulcer in the region of the cardia complicated the tumor. Two sets of conventional biopsies failed to detect malignancy, only inflammation and intestinal metaplasia were seen in the specimens of the mucosa surrounding the ulcer. The endoscopic ultrasonographic findings were an indistinct margin, hypoechogenicity, homogeneous appearance and location within the second and third echographic layer. The surgical resection of the tumor was complemented by an anterior partial fundoplication. The histologic study revealed an inflammatory fibroid polyp, which is a rare, benign, non-capsulated submucosal lesion composed mainly of loose connective tissue and vessels, with an eosinophilic inflammatory component. This lesion is seldom found in the esophagus. PMID- 11005338 TI - Mismatched clinicopathological response after concurrent chemoradiotherapy for thoracic esophageal cancer. AB - We have been treating patients with operable thoracic esophageal cancer according to our own protocol. It includes the initial concurrent chemoradiotherapy (CRT) followed by continuous CRT or surgery. Patients with good response to initial chemoradiotherapy were allowed to continue chemoradiotherapy, whereas the others were treated with surgery. However, there were two cases which showed discrepancies in the clinicopathological response. Both patients received initial chemoradiotherapy, including two courses of cisplatin (100-120 mg), 5 fluorouracil (750-1000 mg for 4 days) and radiation (44-50 Gy). On completion of the initial chemoradiotherapy, all diagnostic imaging modalities including barium swallow, esophagoscopy, endoscopic ultrasonography and thoracic computed tomography strongly implicated residual tumor with a reduction rate of 40-50%. The patients underwent radical esophagectomy 15-20 days after initial chemoradiotherapy. Pathological specimens only revealed thickening of the esophageal wall due to inflammatory change without residual carcinoma. These facts suggest the current limitations of diagnostic images in evaluating the response to chemoradiotherapy. PMID- 11005339 TI - Recurrent gastric carcinoma causing pseudoachalasia: case report. AB - We report the case of a patient with a gastric remnant relapse of an antral carcinoma resected 5 years before and presenting with the clinical feature of a secondary achalasia (pseudoachalasia). In spite of the patient's 4-month history of dysphagia and weight loss that suggested a malignant lesion, barium swallow, repeated endoscopic biopsies and computed tomography (CT) scan of the upper abdomen did not reveal any abnormalities to indicate a recurrence. However, in the following months, because of worsening symptoms, a further CT scan was performed and revealed thickening of the cardia and gastric wall. The patient underwent an exploratory laparotomy that showed an unresectable lesion involving the gastric fundus, the diaphragm and penetrating into the mediastinum, and therefore a palliative jejunostomy was performed. PMID- 11005340 TI - Confronting the problem of increasing antibiotic resistance. AB - Significant increases in prevalence of resistance to antibiotics have been observed in common pathogens of humans in the United States and worldwide. The consequences of the appearance and spread of antibiotic resistance have included increasing morbidity, mortality, and cost of health care. The fundamental cause for the appearance and spread of antimicrobial resistance has been increasing antimicrobial use. However, other factors contribute in both inpatient and outpatient settings. Recognizing the important causes of increasing antibiotic resistance in these settings has led to practical recommendations, which health care facilities and outpatient practitioners will need to review, adapt, and apply for maximum local effectiveness for progress to be made in addressing one of the most challenging problems facing modern medicine. PMID- 11005341 TI - Gambling in the south: implications for physicians. AB - Three historical cycles of legalized gambling have occurred in the South. Currently, every southern state has legalized some form of gaming. Adult past year prevalence rates of problem gambling in southern states are within the national range. Higher prevalence rates occur in states with casinos and multiple forms of legalized gambling. States with lotteries have higher prevalence rates of adolescent problem gambling. Problem gambling can cause stress-induced physical diseases and psychiatric symptoms in gamblers and their families. Physicians can reduce personal, family, and social costs of problem gambling through increased awareness, strategic screening, and early intervention. Treatment approaches include inpatient treatment centers, self-help fellowship groups, and cognitive-behavioral and addiction-based psychotherapies. Although no standard pharmacologic treatments for gambling disorders exist, use of selective serotonin re-uptake inhibitors is under investigation. Referral resources are available to physicians in states with state-funded treatment programs for problem gamblers and/or state councils for problem gambling. PMID- 11005342 TI - The intrauterine device: still misunderstood after all these years. AB - The intrauterine device (IUD) is one of the most effective, safe, and economic methods of contraception today. It is used by more women worldwide than any other reversible method of birth control, yet less than 1% of women in the United States use the IUD. Still remembering the tragedy of the notorious Dalkon Shield, American pharmaceutical companies and clinicians have been wary about promoting any IUD, leaving most physicians inadequately trained in the two IUDs available in the United States. Most significantly, misconceptions about the IUD's safety, mechanism of action, complication rates, and liability have severely limited the utilization of the IUD by many practitioners and women in the United States who may greatly benefit from it. PMID- 11005343 TI - Combined blunt thoracic aortic and abdominal trauma: diagnostic and treatment priorities. AB - Combined blunt trauma to the thoracic aorta and abdomen challenges the surgeon from a diagnostic and therapeutic standpoint. Appropriately prioritizing diagnostic workup and treatment is critical to assuring patient survival. A management approach that considers the patient's injuries and clinical condition as well as the availability of aortography and cardiac surgery are essential. Patients with blunt aortic injury who are hemodynamically unstable with signs of intra-abdominal injury should have immediate abdominal exploration. Further assessment of the aortic injury and surgical repair can be delayed until after the critical intra-abdominal bleeding has been addressed. The stable patient who has both blunt abdominal trauma and blunt thoracic aortic injury but has no signs of ongoing abdominal hemorrhage should initially have arch aortography. Additional abdominal diagnostic studies may be done but should not delay indicated surgical repair of the aortic injury. PMID- 11005344 TI - Dueling doctors. AB - Formal "duels of honor" ruled upper-class conduct for centuries. That social pressure could compel a choice of "death before dishonor" seems nearly incomprehensible today. Physicians might have been expected to disdain dueling as contrary to professional ethics, but a few rather colorful physicians did enter the lists. Details of some of the most famous physician duels are summarized in this article. PMID- 11005345 TI - Striae gravidarum as a predictor of vaginal lacerations at delivery. AB - BACKGROUND: Abdominal stretch marks found during pregnancy may be indicative of poor skin elasticity. One who does not have stretch marks may have better skin elasticity and may be less likely to tear perineal and vaginal tissue during vaginal delivery. This study was conducted to determine whether striae gravidarum could predict lacerations and their severity. METHODS: This prospective observational study included 168 women having vaginal delivery of infants who weighed more than 2,000 g. The absence or presence and degree of lacerations involving the perineum, vagina, labia, and periurethral regions were studied with a step-wise multivariate logistic regression analysis. RESULTS: Episiotomy was found to prevent spontaneous lacerations. Abdominal stretch marks were found to be statistically significant predictors of lacerations when controlling for episiotomy. CONCLUSIONS: Patients with striae gravidarum are at higher risk for lacerations at the time of vaginal delivery than patients who do not have abdominal stretch marks. PMID- 11005346 TI - Tuberculosis exposure in a day-care center: recommended management. AB - BACKGROUND: This article describes the contact investigation, clinical evaluation, and prophylactic treatment of the children and staff members in a day care center after exposure to a teaching assistant with tuberculosis (TB). METHODS: The investigation included baseline and follow-up tuberculin skin testing, history and physical examinations, and chest radiographs. Directly observed prophylactic therapy with isoniazid was recommended for all exposed children with a negative evaluation and all children with positive skin test reactions and normal chest radiographs. Treatment was discontinued in children with negative skin test results at follow-up. Adult staff members were screened with baseline and follow-up skin tests and were referred for chest radiographs if skin test reactions were positive. RESULTS: In 3 of the 141 exposed children and 3 of the 41 adult staff members, tuberculin skin tests yielded positive results. No active TB disease was found. CONCLUSIONS: Isoniazid prophylaxis, administered by directly observed prophylactic therapy in the children, was effectively administered and tolerated in the day-care setting. PMID- 11005347 TI - Use of misoprostol for cervical ripening. AB - BACKGROUND: Misoprostol, the prostaglandin E1 analog, is increasingly used for cervical ripening and induction of labor. We evaluated our experience with misoprostol in an open-label setting. METHODS: Patients were selected for cervical ripening based on clinical profile. At 3 cm cervical dilation, misoprostol was discontinued and other means of labor augmentation were used. Over 13 months, 470 inductions of labor occurred, and 455 charts were available; 254 patients (56%) received misoprostol for cervical ripening, and 144 (32%) received dinoprostone (prostaglandin E2). RESULTS: With misoprostol, mean time from beginning of contractions until delivery was 7 hours, 30 minutes; vaginal birth occurred in 85% of cases, and spontaneous labor occurred in 38%. Hyperstimulation occurred in 4 cases (1.6%) and precipitate labor in 7 (3%). All infants were discharged in excellent condition; one had a 5-minute Apgar score <7, and 33 (13%) had meconium, none with aspiration. Twenty-three patients who had had a previous cesarean section received misoprostol and delivered vaginally. CONCLUSION: Misoprostol was found to be a safe and effective agent for cervical ripening as part of labor induction. PMID- 11005348 TI - Transcutaneous lumbar diskectomy for internal disk derangement: a new indication. AB - BACKGROUND: Percutaneous diskectomy has been used effectively to treat lumbar disk herniation. The purpose of this review was to determine whether transcutaneous diskectomy is an effective surgical option for treating lumbar internal disk derangement. METHODS: Comprehensive retrospective review via personal interview included 103 consecutive patients available for long-term follow-up. For determining results as excellent, good, fair, or poor, the criteria included subjective relief of back and/or leg pain, ability to perform preinjury job functions, residual physical restrictions, use of medications, and need for subsequent surgery. Factors affecting results were statistically analyzed. RESULTS: The overall success rate was 83%, with no statistically significant differences in results based on sex, workers' compensation status, or levels of diskectomy. CONCLUSIONS: The favorable results from this series illustrate the high success rate of transcutaneous lumbar diskectomy in a heterogeneous patient population. Coupling the rate of success with a low rate of complications, transcutaneous lumbar diskectomy appears to be an effective, minimally invasive treatment for internal disk derangement. PMID- 11005349 TI - Pseudomyxoma peritonei manifesting as intestinal obstruction. AB - Pseudomyxoma peritonei is an appendiceal tumor with distinct clinical and pathologic features. It frequently presents a problem in diagnosis and management. We report a case of pseudomyxoma peritonei, which initially appeared with intestinal obstruction. This is a rare initial manifestation in patients who have not had multiple surgical procedures. We review the literature and discuss the unique clinical features and misconceptions surrounding pseudomyxoma peritonei. PMID- 11005351 TI - Secondary abdominal pregnancy in a Jehovah's Witness. AB - A 35-year-old woman, gravida 2, para 1, aborta 0, arrived at our emergency department with abdominal pain of more than 2 weeks' duration. Diagnostic pelvic ultrasonography confirmed a 16-week intra-abdominal pregnancy. Hemoglobin level was 6.9 mg/dL, and hematocrit value was 20.1%. The patient refused blood transfusion on religious grounds. Laparotomy revealed 2,000 mL of blood in the abdomen and a live fetus, with the placenta attached to the omentum and the serosal surface of the right fallopian tube. Postoperative hemoglobin level was 2.8 mg/dL. The patient was transferred to another facility for hyperbaric oxygen therapy, where she subsequently died. Abdominal pregnancy is rare, but has high fetal and maternal mortality rates. Our patient's case was complicated, since she was a Jehovah's Witness and refused lifesaving treatment on religious grounds. Serious medical decisions were made, while respecting the autonomy of the patient. PMID- 11005350 TI - Hypercalcemia and T-cell lymphoma with acquired immunodeficiency syndrome: occurrence without human T-cell leukemia virus-I. AB - We describe the case of a patient with acquired immunodeficiency syndrome (AIDS) who had a CD4 cell count of 60/microL, bilateral hilar adenopathy, and hypercalcemia. Transbronchial biopsy showed T-cell anaplastic large cell lymphoma. Serology was negative for human T-cell leukemia virus-I (HTLV-I). This appears to be the first case of T-cell anaplastic large cell lymphoma occurring in an AIDS patient with hypercalcemia who was seronegative for HTLV-I. PMID- 11005352 TI - Reemergence of sucrose nephropathy: acute renal failure caused by high-dose intravenous immune globulin therapy. AB - Sucrose nephropathy was first described more than 50 years ago. The disorder is characterized by acute renal failure caused by the uptake of sucrose by renal proximal tubule cells with subsequent cellular swelling and occlusion of the tubule lumen. Approximately 114 cases of renal failure have been associated with high-dose intravenous immune globulin (IVIG) therapy. Almost all cases were caused by preparations containing large amounts of sucrose. Clinicians should consider using IVIG preparations containing no sucrose in patients who are at high risk for renal failure. Risk factors include older age, baseline renal failure, and volume contraction. PMID- 11005353 TI - Traumatic asphyxia complicated by unwitnessed cardiac arrest. AB - We report a case of traumatic asphyxia complicated by unwitnessed cardiac arrest in which the patient has made a good, functional recovery. Traumatic asphyxia is an uncommon clinical syndrome usually occurring after chest compression. Associated physical findings include subconjunctival hemorrhage and purple-blue neck and face discoloration. These facial changes can mimic those seen with massive closed head injury; however, cerebral injury after traumatic asphyxia usually occurs due to cerebral hypoxia. When such features are observed, the diagnosis of traumatic asphyxia should be considered. Prompt treatment with attention to the reestablishment of oxygenation and perfusion may result in good outcomes. PMID- 11005354 TI - Cocaine-associated ischemic colitis. AB - Cocaine use can result in various gastrointestinal complications, including gastric ulcerations, retroperitoneal fibrosis, visceral infarction, intestinal ischemia, and gastrointestinal tract perforation. We report cocaine-associated colonic ischemia in three patients and review the literature. Including ours, 28 cases have been reported, with a mean patient age of 32.6 years (range, 23 to 47 years); 53.5% were men and 46.5% were women. The interval between drug ingestion and onset of symptoms varied from 1 hour to 2 days. Cocaine is a potentially life threatening cause of ischemic colitis and should be included in the differential diagnosis of any young adult or middle-aged patient with abdominal pain and bloody diarrhea, especially in the absence of estrogen use or systemic disorders that can cause thromboembolic events, such as atrial fibrillation. PMID- 11005355 TI - Agenesis of the gallbladder. AB - Agenesis of the gallbladder is rare. Three groups have been identified: those with multiple fetal anomalies, asymptomatic cases, and symptomatic cases. Right upper quadrant pain is present in 90% of the cases, nausea and vomiting in 60%, and jaundice in 35%. Operative strategy is aimed at thorough exploration to locate an aberrant gallbladder. We treated a 72-year-old woman with right upper quadrant pain, nausea, and vomiting but no fever or jaundice. Physical examination revealed right upper quadrant tenderness without rebound. The white blood cell count was 10,300/mm3. Total bilirubin level was 1.6 mg/dL. Ultrasonography revealed one gallstone and an enlarged common bile duct. Laparoscopic cholecystectomy was converted to open technique after failure to locate the gallbladder. On intraoperative cholangiogram, no gallbladder was identified. A T-tube was placed. PMID- 11005356 TI - Acute tumor lysis syndrome with choriocarcinoma. AB - A 52-year-old man with retroperitoneal nodal, lung, and liver metastases from choriocarcinoma received chemotherapy with etoposide, cisplatin, and bleomycin. Within 48 hours of starting treatment, he had hypotension, hypoxemia, and anuria. Laboratory values showed hyperuricemia, hyperkalemia, hyperphosphatemia, hypocalcemia, and metabolic acidosis. He was placed on mechanical ventilation, and hemodialysis was instituted, with marked improvement in renal function. A second, shortened course of chemotherapy with carboplatin and etoposide was given 21 days later. However, on hospital day 48, the patient died of progressive pulmonary insufficiency and cardiac arrest. This represents the first reported case of acute tumor lysis syndrome after systemic chemotherapy for advanced nonseminomatous germ cell cancer. PMID- 11005357 TI - Primary nodal neuroendocrine (Merkel cell) tumor in a patient with HIV infection. AB - Lymphadenopathy in the human immunodeficiency virus (HIV) can be of diverse etiology, ranging from infection to cancer. A neoplasm of epithelial origin manifested as inguinal lymphadenopathy without a primary lesion is rare. We report a case of Merkel cell tumor confined only to a lymph node in a patient with the acquired immunodeficiency syndrome (AIDS). We believe this is the first report of primary nodal Merkel cell tumor in a patient with HIV. Because Merkel cell tumor is a rare skin neoplasm with features suggestive of high malignant potential, it is important to distinguish a primary nodal Merkel cell tumor from malignant metastatic processes on the one hand and relatively benign causes of adenopathy on the other. PMID- 11005358 TI - Pyoderma gangrenosum and myelodysplastic syndrome. AB - Pyoderma gangrenosum (PG) is a painful, often rapidly progressive, ulcerating skin disorder frequently associated with systemic diseases. We report the case of a patient with PG and an anemia. A bone marrow biopsy showed changes consistent with one of the myelodysplastic syndromes, refractory anemia with ringed sideroblasts. Patients with PG and anemia should have bone marrow biopsy if no cause of anemia is readily apparent. PMID- 11005359 TI - Subacute thyroiditis manifesting as fever of unknown origin. AB - Subacute thyroiditis (SAT) usually occurs in women in middle age with a viral prodrome, thyroid or neck tenderness, classic symptoms of thyrotoxicosis, and elevated erythrocyte sedimentation rate (ESR). We report a case in an 81-year-old man who initially had 2 days of fever to 101.2 degrees F, confusion, and bilateral lower extremity weakness. Extensive evaluation was remarkable only for the following laboratory values: thyrotropin (TSH) 0.02 microIU/mL, free thyroxine (FT4) 3.1 ng/dL, free triiodothyronine (FT3) 6.0 pg/mL, and ESR 98 mm/hr. One week later, the patient had persistent fevers to 102 degrees F; no source was found. The fever resolved, and 3 months later the patient had profound hypothyroidism (TSH >44.0 microIU/mL, FT4 0.4 ng/dL, ESR 13 mm/hr). A painless thyroid gland and atypical manifestations of hyperthyroidism are unusual in SAT. When fever is of unknown origin, SAT should be considered even if classic features are absent. PMID- 11005360 TI - Cyclooxygenase-2 inhibitor celecoxib: a possible cause of gastropathy and hypoprothrombinemia. AB - Gastrointestinal side effects from nonsteroidal anti-inflammatory drugs (NSAIDs) result mainly from inhibition of the enzyme cyclooxygenase (COX)-1; it is responsible for the synthesis of prostaglandin E2, which leads to increased mucosal blood flow, increased bicarbonate secretion, and mucus production, thus protecting the gastrointestinal mucosa. In inflammation, COX-2 is induced, causing synthesis of the prostaglandins in conditions such as osteoarthritis and rheumatoid arthritis. Two NSAIDs (celecoxib and rofecoxib) with very high specificity for COX-2 and virtually no activity against COX-1 at therapeutic doses have been approved for clinical use. In trials of celecoxib and rofecoxib, only 0.02% of patients had clinically significant gastrointestinal bleeding, compared to a 1% to 2% yearly incidence of severe gastrointestinal side effects with NSAIDs. Our patient had arthritis of the hips and chronic atrial fibrillation and was on warfarin therapy for stroke prevention; less than a week after starting celecoxib therapy, gastrointestinal bleeding and hypoprothrombinemia occurred. PMID- 11005361 TI - Cutaneous nephrocolonic fistula as a consequence of a kidney stone. AB - We report an unusual case of cutaneous nephrocolonic fistula caused by a renal calculus with perirenal infection. The diagnosis was made by fistulography and computed tomography, after which nephrectomy and resection of the descending colon were successful. We also review the literature on cutaneous nephrocolonic fistulas. PMID- 11005362 TI - Hazardous hedgehogs. AB - The African pygmy hedgehog has recently become a fashionable exotic pet in the United States, particularly in the South. As illustrated by the three cases reported, this mammalian insectivore can be a carrier of fungi that cause human dermatomycoses. The African pygmy hedgehog has also been associated with contact urticaria and human salmonellosis. PMID- 11005363 TI - Body temperature of newborns: what is normal? AB - Medical records of 203 healthy full-term infants were reviewed to determine the range of axillary temperatures for newborn infants, factors that affect temperature and nursery management of infants with temperatures outside published normal ranges. The mean birth temperature was 36.5 degrees C (S.D. = 0.6 degrees C). Temperature was associated with birth weight (p<0.0005) and the presence of maternal fever (p<0.0001) but not with type of environment or time of birth. The mean temperature increased with age, rising 0.2 degrees C by 2-3 hours after birth (p<0.0001) and 0.3 degrees C by 15-20 hours (p<0.0001). Among a subset of 114 eligible neonates the mean temperature dropped 0.2 degrees C after bathing (p<0.0001). Although 17% of all temperatures measured were in the hypothermic (< or =36.3 degrees C) range, the only response recorded by nursery staff consisted of warming by modifying the environment, e.g., bundling. Blood cultures were drawn from 51 infants (25%), 43 because of maternal intrapartum antibiotic treatment for maternal fever or prolonged duration of ruptured amniotic membranes (>24 hours) and none for evaluation of abnormal temperatures. No infants had systemic infections and all were discharged in stable condition. Newborn axillary temperatures in our nursery were considerably lower than what has been previously described as "normal." Given the frequency of "hypothermia" and absence of associated illness, we believe the reference range for newborn temperatures should be expanded to include lower temperatures. PMID- 11005364 TI - Significance of a cardiac murmur as the sole clinical sign in the newborn. AB - A cardiac murmur is a very common finding in the first few days of life. It is traditionally believed that lesions creating left-to-right shunts do not present so early. This study was aimed to define and to classify the causes of a murmur in a newborn with an otherwise normal examination. All echocardiograms performed on newborns aged 1-5 days who were referred for evaluation of a murmur in a 3 year period were reviewed. Newborns with additional clinical signs or antenatal diagnosis of a cardiac disease were excluded. Of 20,323 live births, there were 170 newborns referred for echocardiogram solely because of a murmur. Of these, 147 (86%) were found to have structural heart defects. The most common lesions found were those creating left-to-right shunts (66%). Ventricular septal defect was the most common single lesion (54/147, 37%), followed by patent ductus arteriosus (34 newborns, 23%). The combination of both was found in 10 newborns (7%). Six newborns (4%) had pulmonary valve stenosis and three (2%) aortic valve stenosis. Seven newborns (5%) had unforeseen complex heart disease. For five of them, delayed diagnosis would have resulted in potentially life-threatening conditions. There was no correlation between the category of lesion and the age of presentation. The audible threshold of a murmur correlated with a maximum instantaneous gradient of 25 mm Hg (range 11-46 mm Hg). Thirteen percent of newborns with isolated heart murmur had no identifiable structural heart disease. These data suggest that most asymptomatic newborns presenting with a murmur in the first days of life have structural heart disease. Some of the more serious defects would not have been diagnosed without early echocardiography. A left-to right shunt, particularly a ventricular septal defect, may cause a murmur even the first day of life and is probably more common that has been acknowledged. PMID- 11005365 TI - Risk behaviors and the prevalence of Chlamydia in a juvenile detention facility. AB - The prevalence of Chlamydia trachomatis, a sexually transmitted disease, and the association of selected behavioral risk factors were assessed in a population of adolescents in two juvenile detention facilities. Urine was collected for Chlamydia testing and a brief interview conducted for risk and demographic information. Almost all of the 200 participants were sexually active, many before the age of 13 years. The prevalence of Chlamydia infection was 22.2% among female and 8.7% among male participants. Injection drug use was the only statistically significant variable associated with infection. Education and community services are necessary to break the cycle of infection for this high-risk population. PMID- 11005366 TI - Syndrome of hypokalemic metabolic alkalosis and hypomagnesemia associated with gentamicin therapy: case reports. AB - Nephrotoxicity, as evidenced by renal insufficiency is a well-known consequence of gentamicin therapy. We report two patients with gentamicin-induced syndrome of hypokalemic metabolic alkalosis and hypomagnesemia. Both had complete recovery of renal tubular function after cessation of antibiotic therapy. These cases emphasize the need to routinely monitor patients receiving gentamicin therapy for electrolyte abnormalities to avoid potential morbidity. PMID- 11005367 TI - The value of book distribution in a clinic-based literacy intervention program. AB - The purpose of this study was to determine whether anticipatory guidance at well child visits (WCV) that included early literacy development and the provision of books by the examining physician changed family literacy practices. It was conducted in an inner-city pediatric clinic that serves as the continuity practice site for pediatric and pediatric/internal medicine residents. There were 352 children (181 treatment: 171 control), aged 2 to 24 months, enrolled in this prospective, controlled study. The health care providers underwent training on literacy and on how to incorporate this information during WCV. Anticipatory guidance on safety, development, and early literacy was given to all parents. Additionally, the treatment group received an age-appropriate book at each WCV. There were 1,263 visits made (686 treatment, 577 control). Questionnaires were completed by parents on physician helpfulness and by physicians on parental receptiveness. Parental ratings on physician helpfulness were higher in the treatment group than in the control group (p<0.05). Physician's rating of parental receptiveness was also higher in the treatment group than in the control group (p<0.05). Two years after enrollment, mother-child pairs who received guidance and a book were two times more likely to report enjoyment in reading together than the controls who received guidance but no book. We conclude that anticipatory guidance that included early literacy development and distribution of books at WCV resulted in increased family literacy orientation, parental receptiveness, and perception of physician helpfulness. PMID- 11005368 TI - For Michael: those who can, teach. PMID- 11005369 TI - Cough, rash, and dysuria in a preadolescent female. PMID- 11005370 TI - Congenital cholesteatoma of the middle ear. PMID- 11005371 TI - Fever in burned children and its association with infectious complications. PMID- 11005372 TI - Intractable hypoglycemia in a patient with diabetes mellitus, bilateral nephrectomy, and chronic active hepatitis. PMID- 11005373 TI - Introducing Professor John H. Crowe, Anhydrobiosis and Biostabilization Section Editor. PMID- 11005375 TI - Expression of antisense hsp70 is a major determining factor in heat-induced cell death of P-19 carcinoma cells. AB - Overexpressed heat shock protein 70 (Hsp70) is known to be associated with thermoprotection in a number of cell lines and transgenic animals. We hypothesized that because overexpression of Hsp70 protects cells from lethal heat stress, inhibition of expression should make cells susceptible to heat stress. The model used for this study was a stably transfected P-19 carcinoma cell line expressing antisense hsp70 under the control of the hsp70b promoter. The results showed marked inhibition of Hsp70 expression after heat shock correlated with heat-induced cell death. Hsp90 and Hsc70 protein expression were not affected by the antisense construct. Unexpectedly, heme oxygenase (HO-1), another highly inducible heat shock protein, was not induced after heat shock in the antisense hsp70 cell line. Heat shock transcription factor-1 (HSF-1) was in a highly phosphorylated state in the antisense cell line before and after heat shock. This was in contrast to the untransfected control P-19 cells where HSF-1 was primarily highly phosphorylated after heat shock. A control cell line expressing only the vector, pMAMneo, without the antisense construct also showed partial loss of Hsp70 induction but not increased cell death after heat shock. The findings support the role of Hsp70 in thermoresistance. PMID- 11005374 TI - GroEL binds a late folding intermediate of phage P22 coat protein. AB - GroEL recognizes proteins that are folding improperly or that have aggregation prone intermediates. Here we have used as substrates for GroEL, wildtype (WT) coat protein of phage P22 and 3 coat proteins that carry single amino acid substitutions leading to a temperature-sensitive folding (tsf) phenotype. In vivo, WT coat protein does not require GroEL for proper folding, whereas GroEL is necessary for the folding of the tsf coat proteins; thus, the single amino acid substitutions cause coat protein to become a substrate for GroEL. The conformation of WT and tsf coat proteins when in a binary complex with GroEL was investigated using tryptophan fluorescence, quenching of fluorescence, and accessibility of the coat proteins to proteolysis. WT coat protein and the tsf coat protein mutants were each found to be in a different conformation when bound to GroEL. As an additional measure of the changes in the bound conformation, the affinity of binding of WT and tsf coat proteins to GroEL was determined using a fluorescence binding assay. The tsf coat proteins were bound more tightly by GroEL than WT coat protein. Therefore, even though the proteins are identical except for a single amino acid substitution, GroEL did not bind these substrate polypeptides in the same conformation within its central cavity. Therefore, GroEL is likely to bind coat protein in a conformation consistent with a late folding intermediate, with substantial secondary and tertiary structure formed. PMID- 11005376 TI - Cycloheximide- and puromycin-induced heat resistance: different effects on cytoplasmic and nuclear luciferases. AB - Inhibition of translation can result in cytoprotection against heat shock. The mechanism of this protection has remained elusive so far. Here, the thermoprotective effects of the translation inhibitor cycloheximide (CHX) and puromycin were investigated, using as reporter firefly luciferase localized either in the nucleus or in the cytoplasm. A short preincubation of O23 cells with either translation inhibitor was found to attenuate the heat inactivation of a luciferase directed into the cytoplasm, whereas the heat sensitivity of a nuclear-targeted luciferase remained unaffected. After a long-term CHX pretreatment, both luciferases were more heat resistant. Both the cytoplasmic and the nuclear luciferase are protected against heat-induced inactivation in thermotolerant cells and in cells overexpressing heat shock protein (Hsp)70. CHX incubations further attenuated cytoplasmic luciferase inactivation in thermotolerant and in Hsp70 overexpressing cells, even when Hsp70-mediated protection was saturated. It is concluded that protection by translation inhibition is unlikely due to an increase in the pool of free Hsps normally engaged in translation and released from the nascent polypeptide chains on the ribosomes. Rather, a decrease in nascent chains and thermolabile polypeptides may account for the heat resistance promoted by inhibitors of translation. PMID- 11005377 TI - Attenuation of sepsis-induced apoptosis by heat shock pretreatment in rats. AB - Apoptosis is a process by which cells undergo a form of non-necrotic cellular suicide. Although it is a programmed process, apoptosis can be induced by various stressors. During sepsis, apoptosis has been regarded as an important cause of cell death in the immune system, leading to unresponsiveness to treatment. This study was designed to investigate how prior heat shock induction can influence the rate of apoptosis in animals that have experienced sepsis. Sprague-Dawley rats were used, and experimental sepsis was induced by cecal ligation and puncture (CLP). Animals in the heated group were anesthetized and received heat shock by whole-body hyperthermia. They were sacrificed 9 h and 18 h after CLP as early and late sepsis, respectively. Apoptosis was evaluated by "DNA ladder" detection in agarose electrophoresis and Tdt-mediated dUTP nick end-labeling (TUNEL) assay. Hsp72 was detected by Western blot analysis. The results showed that the DNA ladder was detected most clearly in the thymus at the late phase of sepsis with time course dependence, while it showed less clearly in heat shock treated animals. Histopathological study by TUNEL assay obtained similar results in the thymus, where the cortex was more susceptible to apoptosis than the medulla. The Western blot analysis showed that the heat shock induced Hsp72 concomitant with an increase in Bcl-2:Bax ratio. In conclusion, heat shock pretreatment prevents rats from sepsis-induced apoptosis that may account for the better outcome of experimental sepsis. An increase in the Bcl-2:Bax ratio may in part explain the molecular mechanism of the effect of heat shock pretreatment. PMID- 11005378 TI - Characterization of the mouse gene for the heavy metal-responsive transcription factor MTF-1. AB - MTF-1 is a zinc finger transcription factor that mediates the cellular response to heavy metal stress; its targeted disruption in the mouse leads to liver decay and embryonic lethality at day E14. Recently, we have sequenced the entire MTF-1 gene in the compact genome of the pufferfish Fugu rubripes. Here we have defined the promoter sequences of human and mouse MTF-1 and the genomic structure of the mouse MTF-1 locus. The transcription unit of MTF-1 spans 42 kb (compared to 8.5 kb in Fugu) and is located downstream of the gene for a phosphatase (INPP5P) in mouse, human, and fish. In all of these species, the MTF promoter region has the features of a CpG island. In both mouse and human, the 5' untranslated region harbors conserved short reading frames of unknown function. RNA mapping experiments revealed that in these two species, MTF-1 mRNA is transcribed from a cluster of multiple initiation sites from a TATA-less promoter without metal responsive elements. Transcription from endogenous and transfected MTF-1 promoters was not affected by heavy metal load or other stressors, in support of the notion that MTF-1 activity is regulated at the posttranscriptional level. Tissue Northern blots normalized for poly A+ RNA indicate that MTF-1 is expressed at similar levels in all tissues, except in the testes, that contain more than 10 fold higher mRNA levels. PMID- 11005379 TI - Heat shock and arsenite induce expression of the nonclassical class I histocompatibility HLA-G gene in tumor cell lines. AB - The nonclassical histocompatibility class I gene HLA-G has a tissue-restricted expression. To explore mechanisms involved in HLA-G transcriptional regulation, we have investigated the effect of stress, including heat shock and arsenite treatment, on HLA-G expression in tumor cell lines. We show that stress induces an increase of the level of the different HLA-G alternative transcripts without affecting other MHC class I HLA-A, -B, -E, and -F transcripts. A heat shock element (HSE) that binds to heat shock factor 1 (HSF1) on stress conditions was further identified within the HLA-G promoter. Considering the ability of HLA-G to modulate the function of immunocompetent cells, we hypothesize a new feature of HLA-G as a signal regulating the immune response to stress. PMID- 11005380 TI - Formation of nuclear HSF1 granules varies depending on stress stimuli. AB - In concert with the stress-induced activation of human heat shock factor 1 (HSF1), the factor becomes inducibly phosphorylated and accumulates into nuclear granules. To date, these processes are not fully understood. Here, we show that although stress caused by the proteasome inhibitors MG132 and clasto-lactacystine beta-lactone induces the expression of Hsp70, the formation of HSF1 granules is affected differently in comparison to heat shock. Furthermore, proteasome inhibition increases serine phosphorylation on HSF1, but to a lesser extent than heat stress. Our results suggest that, depending on the type of stress stimulus, the multiple events associated with HSF1 activation might be affected differently. PMID- 11005381 TI - Potential targets for HSF1 within the preinitiation complex. AB - Protein-protein interactions between human heat shock transcription factor 1 (hHSF1) and general transcription factors TFIIA-gamma, TFIIB, TBP, TAF(II)32, and TAF(II)55 and positive coactivator PC4 were characterized in order to identify potential targets of contact in the transcriptional preinitiation complex. These contacts represent one of the final steps in the signal transfer of heat stress to the transcriptional apparatus. TATA-binding protein (TBP) and transcription factor IIB (TFIIB) were identified as major targets for HSF1 transcriptional activation domains AD1 and AD2 based on in vitro interaction assays. TBP showed affinity for AD2 and a fragment containing AD1, while the core domain of TFIIB interacted primarily with the AD1 fragment. Interactions were also detected between full-length HSF1 and the small subunit (gamma) of TFIIA. PC4 interacted weakly with HSF2 and showed even less affinity for HSF1. Coimmunoprecipitation of transiently expressed TBP in HeLa cells demonstrated that HSF1 AD2 and AD1+AD2 are able to bind TBP in vivo. Assays based on transcriptional interference confirmed predictions that both TBP and TFIIB can interact with HSF1 activation domains in HeLa cells. The negative regulatory region (NR) of HSF1 did not interact with any general factors tested in vitro but did bind TFIID in nuclear extracts through contacts that probably involve TATA associated proteins (TAFs). These results suggest a model for transcriptional regulation by HSF1 that involves a shift between formation of dysfunctional TFIID complexes with the NR and transcriptionally competent complexes with the C-terminal activation domains. PMID- 11005383 TI - The Escherichia coli heat shock protein ClpB restores acquired thermotolerance to a cyanobacterial clpB deletion mutant. AB - In both prokaryotes and eukaryotes, the heat shock protein ClpB functions as a molecular chaperone and plays a key role in resisting high temperature stress. ClpB is important for the development of thermotolerance in yeast and cyanobacteria but apparently not in Escherichia coli. We undertook a complementation study to investigate whether the ClpB protein from E coli (EcClpB) differs functionally from its cyanobacterial counterpart in the unicellular cyanobacterium Synechococcus sp. PCC 7942. The EcClpB protein is 56% identical to its ClpB1 homologue in Synechococcus. A plasmid construct was prepared containing the entire E coli clpB gene under the control of the Synechococcus clpB1 promoter. This construct was transformed into a Synechococcus clpB1 deletion strain (deltaclpB1) and integrated into a phenotypically neutral site of the chromosome. The full-length EcClpB protein (EcClpB-93) was induced in the transformed Synechococcus strain during heat shock as well as the smaller protein (EcClpB-79) that arises from a second translational start inside the single clpB message. Using cell survival measurements we show that the EcClpB protein can complement the Synechococcus deltaclpB1 mutant and restore its ability to develop thermotolerance. We also demonstrate that both EcClpB-93 and 79 appear to contribute to the degree of acquired thermotolerance restored to the Synechococcus complementation strains. PMID- 11005384 TI - Commentaries on Coburn 'The role of neo-liberalism'. PMID- 11005382 TI - Aryl hydrocarbon (Ah) receptor levels are selectively modulated by hsp90 associated immunophilin homolog XAP2. AB - The aryl hydrocarbon receptor (AhR) is a ligand-inducible transcription factor that mediates biological responses to halogenated aromatic hydrocarbons. The unliganded AhR is a cytoplasmic, tetrameric complex consisting of the AhR ligand binding subunit, a dimer of hsp90, and the hepatitis B virus X-associated protein 2 (XAP2). The role of XAP2 as a member of the AhR core complex is poorly understood. XAP2 shares significant homology with the immunophilins FKBP12 and FKBP52, including a highly conserved, C-terminal, tetratricopeptide repeat (TPR) domain. XAP2 forms a complex with hsp90 and the AhR but can also bind to both independently. This binding is mediated by the conserved TPR domain. Single-point mutations in this region are sufficient to disrupt the association of XAP2 with both the AhR and hsp90 in cells. Cotransfection of the AhR and XAP2 in COS-1 cells results in increased AhR levels compared with cells transfected with the AhR alone. In contrast, coexpression of the AhR with the TPR containing proteins FKBP52, protein phosphatase 5 (PP5), or XAP2 TPR-mutants deficient in binding to the AhR and hsp90 does not affect AhR levels and coexpression of the AhR with the TPR domain of PP5 results in AhR down-regulation. These results demonstrate that XAP2 is apparently unique among hsp90-binding proteins in its ability to enhance AhR levels. A yellow fluorescent protein (YFP)-XAP2-FLAG was constructed and biochemically characterized, and no loss of function was detected. YFP-XAP2-FLAG was transiently transfected into NIH 3T3 and was found to localize in both the nucleus and the cytoplasm when visualized by fluorescence microscopy. Treatment of Hepa-1 cells with the hsp90-binding benzoquinone ansamycin, geldanamycin, and the macrocyclic antifungal compound radicicol resulted in AhR but not XAP2 or FKBP52 turnover. Taken together, these results suggest that XAP2/hsp90 and FKBP52/hsp90 complexes are similar yet exhibit unique functional specificity. PMID- 11005385 TI - Coburn's thesis: plausible, but we need more evidence and better measures. PMID- 11005386 TI - Deeper than "neoliberalism". A reply to David Coburn. PMID- 11005387 TI - Income inequality and health: expanding the debate. PMID- 11005388 TI - Social change, market forces and health. PMID- 11005389 TI - Unintended pregnancy and women's use of prenatal care in Ecuador. AB - This paper assesses the relationship between unintended pregnancy--both unwanted and mistimed and several dimensions of use of prenatal care among women in Ecuador, where the level of unintended pregnancy has risen considerably in recent years. Data were collected from a nationally representative sample of 3988 women interviewed in the 1994 Demographic and Maternal-Child Health Survey. Multivariate logistic regression was used to assess jointly the effect of pregnancy intention status (unwanted, mistimed, planned) on three aspects of prenatal care use while controlling for potential confounders. Women with unwanted pregnancies were 32% less likely than women with planned pregnancies to seek out prenatal care. Women with unwanted pregnancies were also 25% less likely to initiate care in the first trimester and 29% less likely to receive at least an adequate number of visits. Mistimed pregnancy was not associated with receiving care, timely initiation of care or receiving an adequate number of visits. PMID- 11005390 TI - Social class and self-rated health: can the gradient be explained by differences in life style or work environment? AB - The purpose of the present paper is to describe differences in work environment and life style factors between social classes in Denmark and to investigate to what extent these factors can explain social class differences with regard to changes in self-rated health (SRH) over a 5 year period. We used data from a prospective study of a random sample of 5001 Danish employees, 18-59 years of age, interviewed at baseline in 1990 and again in 1995. At baseline we found higher prevalence in the lower classes of repetitive work, low skill discretion, low influence at work, high job insecurity, and ergonomic, physical, chemical, and climatic exposures. High psychological demands and conflicts at work were more prevalent in the higher classes. With regard to life style factors, we found more obese people and more smokers among the lower classes. The proportion with poor SRH increased with decreasing social class at baseline. The follow-up analyses showed a clear association between social class and worsening of SRH: The lower the social class, the higher the proportion with deterioration of SRH. There was no social gradient with regard to improved SRH over time. Approximately two thirds of the social gradient with regard to worsening of SRH could be explained by the work environment and life style factors. The largest contribution came from the work environment factors. PMID- 11005391 TI - "The lizard in the green bottle": "aging out" of problem drinking among Navajo men. AB - The Navajo exhibit a number of indicators suggesting the extent of significant problems associated with drinking and alcohol abuse. Measures of alcohol-related mortality and morbidity provide stark testimony regarding the shape and magnitude of problem drinking among the Navajo. While these measures highlight patterns of drinking that often result in social, physical, and psychological pathology, there are other, less noted patterns of Navajo drinking. This paper describes a salient, if often overlooked, pattern of Native American drinking by examining the aging out phenomenon among Navajo men. Using narratives collected from former problem drinkers, this paper describes the factors and motivations associated with this sometimes dramatic change in drinking behavior. Several themes emerge from these narratives that help explain the aging out process. These themes include health concerns, religious involvement, living a traditional Navajo way of life, and the responsibilities and life changes associated with child rearing. This paper not only provides detail on a category of drinker and drinking phenomena that is largely ignored in accounts of Native American drinking, but also illustrates some of the values and meanings attached to drinking cessation and highlights the relation between changes in drinking behavior and stages in life course among Navajo men. PMID- 11005392 TI - New Zealand children's health camps: therapeutic landscapes meet the contract state. AB - This paper surveys the history and current status of children's health camps in New Zealand, and places these sites within the theoretical context of therapeutic landscapes. The first health camp was established in 1919, and the seven current camps provide respite, education and health care for approximately 4000 children each year. We analyse the health-place relations inherent in the health camp concept and suggest that the 'therapeutic landscape' idea developed by Gesler provides a useful framework to explain the development of camps as sites for enhancing child and family welfare. Specifically, we contend that changing understandings of health and children have been closely linked with changing perceptions of what is therapeutic about the camps. Survey data demonstrate that contemporary restructuring of the welfare state has recast the role of health camps and placed them in a precarious position in terms of both financial viability and public acceptability. We conclude that the current status of health camps is ambiguous given the pressures of deinstitutionalisation philosophies and the regulatory environment of formal contracts between funders and providers. PMID- 11005393 TI - Adult life experiences and health in early old age in Great Britain. AB - In Britain and other developed countries older people comprise a large majority of all those reporting long term illness or disability. However, most studies of socio-demographic variations in health have focussed on those in younger age groups. Moreover approaches to the study of health variations are often fragmented. In this study we have adopted a life course approach to analyse differentials in health in early old age. The data comes from the Retirement and Retirement Plans Survey and follow-up, a two-wave study of persons aged 55-69 in 1988/9. As well as information on current circumstances, the data set includes occupational, marital, and fertility history information. At baseline a nationally representative sample of the population of Great Britain were interviewed at home by trained interviewers (n = 3543). The sample was followed up and in 1994, 2247 survivors were re-interviewed, a response rate of 70% (of survivors). The data were weighted to adjust for non-response bias. Two outcome measures were used: self rated health and presence or absence of disability assessed from a scale derived from detailed questions on thirteen domains of disability. The severity score used was that developed for the 1985/6 ONS Surveys of Disability. The findings indicate that health and disability status at baseline and at follow up were associated with socioeconomic and geographic variables, such as proportion of adult life spent unemployed and residence outside the Southeast of England; demographic factors, such as early age at marriage and high parity; and experience of adverse events, such as the death of a child and being dismissed from work. The results show that socio-economic, demographic, and geographical and life events' factors are all associated with health status in early old age and that integrated, rather than bifurcated, approaches to the study of health differentials are needed. PMID- 11005395 TI - Patient-centredness: a conceptual framework and review of the empirical literature. AB - A 'patient-centred' approach is increasingly regarded as crucial for the delivery of high quality care by doctors. However, there is considerable ambiguity concerning the exact meaning of the term and the optimum method of measuring the process and outcomes of patient-centred care. This paper reviews the conceptual and empirical literature in order to develop a model of the various aspects of the doctor-patient relationship encompassed by the concept of 'patient centredness' and to assess the advantages and disadvantages of alternative methods of measurement. Five conceptual dimensions are identified: biopsychosocial perspective; 'patient-as-person'; sharing power and responsibility; therapeutic alliance; and 'doctor-as-person'. Two main approaches to measurement are evaluated: self-report instruments and external observation methods. A number of recommendations concerning the measurement of patient centredness are made. PMID- 11005394 TI - Factors associated with non-urgent utilization of Accident and Emergency services: a case-control study in Hong Kong. AB - Accident and Emergency Departments (A&E) have been a popular source of primary care, and studies have shown that up to two thirds of patients attending A&E have problems that could be managed by general practitioners (GPs). Although many studies have found that patients of lower socio-economic class with less social support have a higher utilization rate of A&E, some recent studies have revealed contrary evidence. In this study 2410 patients were randomly selected from four A&E at different times. The gold standard in differentiating true emergency cases and GP cases was based on a retrospective record review conducted independently by a panel of emergency physicians. Two emergency physicians reviewed each case independently, and if their independent ratings were in agreement, this became the gold standard. Patients classified as GP cases were given a telephone interview, and a sample was selected and matched with cases from general out patient clinics (GOPC) in the public sector by morbidity. Reasons for not attending a private GP included closure of clinic, deterioration of symptoms, GPs' inability to diagnose properly, and patients' wish to continue medical treatment in the same hospital. Reasons why non-urgent patients did not choose to attend the nearby public GOPC included affordability, closure of the GOPC, patients' wish to continue treatment at the same hospital, GOPC too far away, no improvement shown after visits to GOPC doctors, and GOPC doctors' inability to make proper diagnoses. The reasons for high level of utilization of A&E services are complex and reflect problems of delivery of GP services. There is an urgent need for GPs to set up a network system to provide out of hours services, and also for a better interfacing between primary and secondary care, and between public and private sectors, so that patients can be referred back to GPs. Interim clinical services provided to those non-urgent cases by nursing practitioners or by GPs working in A&E could also facilitate discharge of patients to primary care facilities. PMID- 11005396 TI - The inertia of self-regulation: a game-theoretic approach to reducing passive smoking in restaurants. AB - Two alternate regulatory approaches can be used to reduce exposure to environmental tobacco smoke in workplaces. The first is voluntary, self regulation introduced by management, which is supported by common law and occupational health legislation that emphasises the employers' 'duty of care'. The second is public health legislation that bans smoking outright in enclosed places. In Australia, self-regulation has succeeded in restricting tobacco smoking in most indoor workplaces but has been a relative failure in the hospitality industry. Claims that this reflects consumer preference by diners, club and hotel patrons are not backed by survey evidence, typically showing large majority support for non-smoking establishments. Insights from game theory show why reliance on the duty of care is unlikely to succeed even when establishment operators collectively support a non-smoking policy. Using plausible assumptions about the net costs of unilaterally introducing smoking restrictions, what makes good sense for society as a whole is likely to be the least profitable option for an individual operator acting alone. Operators find themselves in the classic prisoner's dilemma. If the aim of policy is to restrict smoking in public places in order to protect the health of employees then game-theory predicts that public health legislation banning smoking in enclosed places will be more effective than self-regulation and reliance on the duty of care. PMID- 11005397 TI - Social class differences in mortality using the National Statistics Socio economic Classification--too little, too soon: a reply to Chandola. AB - Chandola's (2000; Social Science and Medicine, 50(5), 641-649) claims that the UK National Statistics Socio-economic Classification (NS-SEC) is not significantly related to mortality are examined. It is shown that this result is likely to be an artefact of the data and methods he uses. Other findings and interpretations in his paper are also critically discussed. In the process, various theoretical, measurement and analysis issues relating to the use of classifications such as the NS-SEC in health research are reviewed. PMID- 11005398 TI - Casualty, accident and emergency, or emergency medicine, the evolution. PMID- 11005399 TI - A problem-based learning resource in emergency medicine for medical students. AB - Emergency medicine is a relatively new specialty area within medicine, however medical schools, students and standard setting bodies have recognised that learning emergency medicine is integral to the training of medical students. There are, however, significant problems with the delivery of emergency medicine teaching including low teacher numbers, severely limited teaching time and lack of suitable learning resources. This paper describes the process of development of a learning resource, its format and content and summarises student feedback. PMID- 11005400 TI - Acute occlusion of the retinal arteries: current concepts and recent advances in diagnosis and management. AB - PURPOSE/BACKGROUND: Central retinal artery occlusion (CRAO) is usually a blinding event, and is not an infrequent presentation to the accident and emergency (A&E) department. The evidence-base in support of current treatment options is weak. METHODS: This paper reviewed the literature germane to the diagnostic, therapeutic and prognostic aspects of retinal arterial occlusive disease. RESULTS: The visual prognosis associated with CRAO remains poor, and current therapeutic practices are of unproven benefit. The non-ophthalmologist in the A&E department should lie the patient flat and give a stat dose of intravenous acetazolamide in an attempt to improve the retinal perfusion pressure. CONCLUSION: The management of acute occlusion of the central retinal artery has not changed over the past 30 years, although the potential benefits of superselective intra-arterial fibrinolytic therapy warrant evaluation in a randomised controlled trial. The identification of underlying pathology is an essential component of medical care, and all cases should be followed up by an ophthalmologist because of the possibility of ocular rubeosis. PMID- 11005401 TI - The focused trauma ultrasound examination. Can, and should, accident and emergency physicians in the UK acquire this skill? PMID- 11005402 TI - Neonatal head injuries. AB - A retrospective case note review of head injuries in neonates admitted to the Neonatal Surgical Unit in Glasgow between 1990 and 1996 (n=25) was carried out. Most injuries were caused by a fall (68%) and resulted in scalp haematomata and associated skull fractures in the majority of patients. Three neonates were involved in high speed road traffic accidents, and these infants all had intracranial pathology identified by computed tomography. Isolated skull fractures were common and did not appear to be associated with any neurological deficit. Nonaccidental injury was uncommon in this age group. Outcome was excellent in the majority of patients (92%). PMID- 11005403 TI - Next day telephone follow up of the elderly: a needs assessment and critical incident monitoring tool for the accident and emergency department. AB - BACKGROUND: Patients over the age of 75 years comprise an increasing proportion of accident and emergency (A&E) department attendances. Within this group there is a high incidence of comorbidity, which mandates effective discharge coordination from the A&E department. OBJECTIVES: The aims of this study were to assess the needs of these patients the day after discharge, target patients for appropriate interventions and identify critical incidents. SETTING: The study was undertaken in a district general hospital A&E department that has 62000 new patient attendances per year. INCLUSION CRITERIA: Patients aged 75 years or over who were discharged from the A&E department. EXCLUSION CRITERIA: Nursing home patients. Patients without a telephone. STUDY DESIGN: Pre-discharge information was collected from the medical notes. A community liaison nurse (CLN) then contacted patients by telephone. A semistructured questionnaire was used to assess patients. Patients were risk stratified and appropriate interventions made. Interventions initiated by the CLN were scored from 1 to 6 based on the level of input required. RESULTS: 551 patients or their carers were contacted by telephone. Existing home support was felt to be insufficient in 44 (8%) cases and in need of immediate intervention in a further 45 (8%) cases. Sixty five (11%) Category 1 patients required no intervention, 223 (42%) Category 2 patients required advice only, 107 (19%) Category 3 patients were referred to their GP, 127 (23%) Category 4 patients required a domicillary visit by a GP or a nurse, 26 (5%) Category 5 patients were at risk requiring urgent home assessment and three Category 6 patients had to re-attend A&E. Advice was given by the CLN on a broad range of issues and a wide range of health care services was accessed. Five hundred and fifty nine referrals were made by the CLN after telephone assessment. CONCLUSIONS: Telephone follow up of patients over 75 attending our A&E department identified a number of areas where care could be improved before and after discharge. This low cost, high quality intervention has the potential for decreasing inappropriate return visits to the department by a vulnerable group of patients as well as improving overall quality of care. PMID- 11005404 TI - Biological tissue adhesive for multiple use in the accident and emergency department. AB - OBJECTIVE: To assess the strength of the glue and microbial contamination over 28 days from opening a vial of tissue adhesive in the accident and emergency setting, and to quantify cost savings of repeated use of the vials. METHOD: (1) Strips of reinforced nylon and a specially constructed piece of apparatus designed to measure the force at which the glue gave way were used to measure the strength of the tissue adhesive at various times after the glue was opened to assess if the glue strength deteriorated over time. (2) Microbial contamination of the glue was assessed. RESULTS: There was no deterioration in the glue strength over time. There was no evidence of microbial contamination of the glue. CONCLUSION: Cyanoacrylate tissue adhesive can safely be reused for a period of 28 days after opening with no risk of degradation of glue strength or contamination with micro-organisms. In our department this represents a potential saving of l5400 per year. PMID- 11005405 TI - The capability of accident and emergency departments to safely decontaminate victims of chemical incidents. AB - OBJECTIVES: To evaluate the capability of accident and emergency (A&E) departments in six health regions of England to safely decontaminate casualties exposed to hazardous chemicals. METHODS: In January 1999 a postal questionnaire was sent to the clinical director of all A&E departments in Trent, North and South Thames, South and West, North West and, Anglia and Oxford Health Regions. The questionnaire inquired about characteristics of the department, decontamination facilities and equipment, and staff training. Nonresponders were sent a second questionnaire and contacted by telephone if they failed to respond to the second mailing. RESULTS: 308 of 326 departments identified (94%) returned a questionnaire. There was no significant difference in response rate by region (p = 0.99). Analysis was restricted to 154 major departments seeing more than 20000 new attendances per year. Of these 154 departments, 109 (71%) had a written chemical incident plan but only 55 (36%) maintained a list of nearby industrial chemical sites. Fifty nine departments (38%) stated that members of staff had received training in the management of chemically contaminated casualties in the preceding year. Eighteen departments (12%) possessed the level of personal protective equipment (PPE) recommended for decontamination by the Ambulance Services Association. Ninety six departments (62%) had a designated decontamination room but only seven (7%) of them incorporated all the features generally considered necessary for safe decontamination. Forty one units (27%) had the capability to decontaminate casualties outside of the department either with warm water from a shower attachment or with a mobile decontamination unit. Thirty six departments (23%) had neither a decontamination room nor the ability to decontaminate casualties outside the department. Only 16 units (10%) had both adequate PPE and either a decontamination room or the capability to decontaminate outside the department. CONCLUSIONS: This study has identified deficiencies in the current NHS capability to respond to chemical incidents. To resolve this, nationally recognised standards for decontamination facilities, equipment and training should be formulated, agreed and implemented. PMID- 11005406 TI - Referral to the accident and emergency department following the use of community alarms. AB - OBJECTIVES: To assess the degree of appropriate referral to the accident and emergency (A&E) department following the use of a community alarm where a mobile warden works in conjunction with the community alarm control centre. METHODS: 611 consecutive calls using community alarms underwent assessment and the appropriateness of referral to the A&E department was considered. RESULTS: Of 542 requests for help 44 patients were transported to the A&E department (8.1%). Twenty nine patients were admitted (5.3%) and 15 patients (2.8%) discharged home from the A&E department after assessment or treatment, or both. Only three patients (0.55%) had been referred to the A&E department inappropriately. CONCLUSIONS: This study shows that where a mobile warden works in conjunction with the community alarm control centre the number of inappropriate referrals to the A&E department should be minimal. PMID- 11005407 TI - Regular attenders to the accident and emergency department. AB - OBJECTIVES: To examine the profile of regular attenders to an accident and emergency (A&E) department, and to estimate the percentage of the overall departmental workload attributed to this group of patients, together with the resultant cost to the department of these attendances. METHODS: A retrospective study of regular attenders to the A&E department at Hull Royal Infirmary was conducted between 1 January 1998 and 30 June 1998. The information gathered included age, sex, marital status, accommodation, investigations performed, concurrent alcohol use, presenting complaints and disposal. RESULTS: The A&E department at Hull Royal Infirmary sees approximately 87 000 new patients per year. Forty regular attenders presented 475 times in six months accounting for 1.1% of the departmental workload. The most common presenting complaints were overdose (27.4%), minor injuries (19%), alcohol intoxication (14%) and seizures (10.5%). Eighty per cent of patients were single and 7.5% were of no fixed abode. A total of 191 admissions resulted and the cost to the department for investigations performed was between l2709.59 and l3739.85. The cost of inhospital admissions was in excess of l34000. CONCLUSION: Improved management of these patients together with a reduction in their alcohol intake may lead to a significant reduction in both workload for accident departments and hospitals and in the number of regular attenders. PMID- 11005408 TI - Questionnaires of accident and emergency departments: are they reproducible? AB - BACKGROUND: Questionnaires are commonly sent to accident and emergency (A&E) departments to determine common practice and are often extrapolated to best practice. AIMS: To determine if questionnaire based studies have a defined population of A&E departments and whether studies are reproducible. METHODS: All questionnaires in the Journal of Accident and Emergency Medicine were reviewed and assessed for inclusion criteria, departments studied and study design. RESULTS: 30 questionnaires were detected, 22 were postal, six telephone and two did not state method of contact. Sample sizes ranged from 15 to 740 and inclusion of A&E departments was highly variable according to geographical area, size of department or consultant status. Seventeen (54.8%) did not state the source of A&E department listings. Response rates ranged from 55-100%. Only three studies undertook subset analysis according to either size or locality. CONCLUSIONS: Questionnaire of studies A&E departments have poor methodology descriptions, which means that many are not reproducible. Inclusion criteria are highly variable and failure to analyse important subsets may mean that individual departments cannot apply recommendations. Questionnaire studies relating to A&E do not use a consistent well defined population of A&E departments. Information in the studies is usually inadequate to allow them to be repeated. PMID- 11005409 TI - An introduction to statistical inference--3. AB - Statistics inference is used to make comments about a population based upon data from a sample. In a similar manner it can be applied to a population to make an estimate about a sample. It is commonly seen in medical publications when the null hypothesis is being tested. This calculates the probability (p value) of a type I error--that is, that a particular finding is attributable to chance. It is also important to be aware of the chances of a type II error--that is, accepting the null hypothesis when it does not apply. Sample size, point estimate and variability are common factors that will affect the chances of making these two types of errors. Interpreting results therefore needs to take these factors into account as well as the clinical relevance of the findings. Statistical significance does not necessarily mean clinical significance. PMID- 11005410 TI - Towards evidence based emergency medicine:Best BETs from the Manchester Royal Infirmary. Use of the McCoy laryngoscope in patients with suspected cervical spine fracture. PMID- 11005411 TI - Towards evidence based emergency medicine:Best BETs from the Manchester Royal Infirmary. White cell count and diagnosing appendicitis in pregnancy. PMID- 11005412 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Oral acyclovir in acute cutaneous herpes zoster. PMID- 11005413 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Urinary trypsinogen to rule out acute pancreatitis in patients with abdominal pain. PMID- 11005414 TI - Journal scan. PMID- 11005415 TI - Injuries sustained by aircrew on ejecting from their aircraft. AB - This paper describes some of the injuries sustained by the aircrew who ejected from their aircraft after a mid-air collision, and discusses the types of injury that such patients may suffer. PMID- 11005416 TI - Adult spinal cord injury without radiological abnormality. AB - Spinal cord injury without radiological abnormality is rare in adults. A case is described of a 61 year old man who fell 15 feet from a ladder striking his head on a wall who presented with neck pain and with motor and sensory neurological abnormalities in his limbs. Plain radiographs of the neck revealed no fractures or dislocations. Further imaging with computed tomography and magnetic resonance imaging revealed an osteophyte fracture with associated cord contusion at the C5 level. Careful neurological examination is essential in all cases of potential spinal injury. PMID- 11005417 TI - Hyperventilation: cause or effect? AB - A young person presenting with shortness of breath is common to the accident and emergency department. Usually this hyperventilation is anxiety related or a panic attack, but sometimes it can be caused by a serious underlying condition like pulmonary embolus. Acute shortness of breath in any patient should never be dismissed lightly. It is important to realise that pulmonary embolus can present without chest pain and with shortness of breath as the major symptom. Such patients can be distinguished by close attention to history and examination, risk factors for thromboembolic disease and the use of basic investigations (electrocardiogram, chest radiography and arterial blood gas analysis). A serious cause for shortness of breath must be excluded before labelling it as "hysteria" or "panic". PMID- 11005418 TI - Combined brachial plexus and vascular injury in the absence of bony injury. AB - Neurovascular injury to the axillary vessels is well described in association with fracture or dislocation involving the shoulder joint or the humerus. Such injury however can also occur in the absence of bony injury. A case is presented of damage to the axillary artery and brachial plexus following blunt trauma. This case demonstrates that complex neurovascular damage can occur in the absence of fracture or dislocation. The importance of a thorough clinical assessment is highlighted and priorities with regard to diagnosis and management are discussed. PMID- 11005419 TI - Acute cerebrovascular accident after minor trauma in a 1 year old. AB - Acute cerebrovascular accident in an otherwise well child is a rare presentation. A case is described where the diagnosis was delayed because of association with minor trauma and a misleading diagnosis of soft tissue injury. PMID- 11005420 TI - Trauma induced testicular torsion: a reminder for the unwary. AB - Trauma induced testicular torsion is a well recognised entity, the incidence being 4-8% in most studies reporting on testicular torsion. The signs and symptoms of testicular torsion may easily be mistakenly attributed to preceding testicular trauma if there was such an event. A patient is described with trauma induced testicular torsion who presented on three occasions before a decision was made to perform scrotal exploration. Unfortunately, an orchidectomy was the outcome. The message that trauma can and not infrequently does precipitate torsion, needs to be reiterated. Awareness of the entity and constant vigilance is required of clinicians to avoid a delay in definitive treatment. PMID- 11005421 TI - Retrieval of a metallic foreign body in the neck with a rare earth magnet. PMID- 11005422 TI - Circ.com-plications. PMID- 11005423 TI - An unusual diagnosis for a swollen limb. PMID- 11005424 TI - An unusual cause of stridor. PMID- 11005425 TI - A case of bilateral Lisfranc fracture--dislocations in a patient following polytrauma. PMID- 11005426 TI - Rabies in the accident and emergency department. PMID- 11005427 TI - Consent in epidemiology: implications of history for public policy. PMID- 11005428 TI - Strontium-90 in newborns and childhood disease. AB - Radioactive strontium-90 concentrations in baby teeth obtained from Suffolk County, New York, rose steadily during the 1980s. Recent levels of strontium-90 are similar to those reported for babies born in the late 1950s-at the height of atmospheric nuclear weapons testing in Nevada. Strontium-90 concentrations increased concomitantly with increases in cancer incidence among Suffolk children under the age of 5 y, a result that mimicked parallel trends observed in the 1950s and early 1960s. Given that effects of strontium-90 on developing cells are most pronounced during the fetal and infant periods, escalating levels should be viewed as a factor in the recent decline in various child health status measures. PMID- 11005429 TI - Blood lead secular trend in a cohort of children in Mexico City. II. 1990-1995. AB - The authors determined the secular trend in blood lead levels in a cohort of 91 children born in Mexico City between 1987 and the beginning of 1993. The authors grouped children by calendar year in which they reached 36 mo of age (i.e., 1990 1995), and their blood lead levels were measured every 6 mo during a 66-mo period. The overall geometric mean blood lead level was 8.6 microg/dl (range = 1.0-61.0 microg/dl). A repeated measures analysis of variance revealed a significant downward linear trend in blood lead levels by year (p < .001)--from an estimated marginal geometric mean of 14.2 microg/dl in 1990 to 6.3 microg/dl in 1995. There was also a significant linear age effect (p < .001); blood lead levels generally fell during the 36th-66th mo. Family use of lead-glazed pottery significantly elevated blood lead levels (p = .006), and the effect magnified as age increased (Age x Pottery Interaction [p = .014]). Although the overall downward trend in blood lead levels during the time period described corresponded to a reduction in various sources of lead exposure, there was no alteration in production, distribution, or use of leaded pottery. Currently, use of lead-glazed ceramic pottery is one of the most profound sources of lead exposure in the Mexican population. PMID- 11005430 TI - A new technology to measure skin absorption of vapors. AB - Skin vapor absorption is one of the major exposure routes for some widely used chemicals (e.g., 2-methoxy ethanol), but a good apparatus with which exposure can be measured is currently unavailable. In this study, a polished stainless-steel chamber-combined with computer-controlled auto-feedback software and hardware, real-time gas sensors, and an auto-injection microsyringe-was proposed as new technology. In addition, the machines had activated-charcoal tubes and cold traps, both of which simulated the skin uptake and validated the reliability of the proposed system. The exposure concentrations, relative humidity, and temperature were effectively controlled at 25+/-0.5 ppm (or 300+/-10 ppm), 80+/ 2%, and 27.5+/-0.5 degrees C, respectively. The relative errors between the quantity of 2-methoxy ethanol collected in either the charcoal tubes or the cold traps and the quantity of ME injected to maintain a constant exposure were less than 5%. The authors also used this new technology to successfully measure skin absorption of ME vapor in 6 volunteers. The authors concluded that this new technology is a direct, continuous, noninvasive, and simple tool with which to measure skin absorption of vapors. PMID- 11005431 TI - Biomonitoring study of people living near or working at a municipal solid-waste incinerator before and after two years of operation. AB - The authors conducted a biomonitoring study in the town of Mataro, Spain, of 104 subjects who lived near (i.e., within 0.5-1.5 km) an incinerator, 97 subjects who lived far (i.e., within 3.5 km-4.0 km) from an incinerator, and 17 workers at a new municipal solid-waste incinerator. The study commenced before the incinerator started functioning in 1995, and 2 y later (1977) the authors undertook the final part of the study. Dioxins, furans, and polychlorinated biphenyls were studied in pooled blood samples (n = 22), and individual blood and urine samples were analyzed for the detection of lead, chromium, cadmium, and mercury. In 1995, dioxin blood levels were low-both among those living close to the incinerator (mean = 13.5 ng international-dioxin toxic equivalents/kg fat) and among those living far away (mean = 13.4 ng international-dioxin toxic equivalents/kg fat). In 1997, dioxin and polychlorinated biphenyl levels had increased in both groups of residents by approximately 25% and 12%, respectively. (The increase in dioxin levels was about 10% when the authors took into account the mean of two repeated quality-control analyses.) Blood lead levels decreased, but no difference was observed for chromium, cadmium, and mercury. Minimal changes were seen among workers. Given the low dioxin stack emissions from this plant (mean = 2.5-0.98 ng international-dioxin toxic equivalents/m3) and that the blood dioxin levels did not depend on distance of residence from the incinerator, it would appear unlikely that the small increase in dioxin blood levels resulted from the incinerator's emissions. PMID- 11005432 TI - Study of the genotoxicity of toluene. AB - Chromosome analysis was conducted for peripheral lymphocytes of 23 printers exposed to toluene concentrations of 590 mg/m3 in a rotary machine workshop and to rotogravure printing inks. The percentages of aberrant cells were 2.30 in the printers and 1.46 in the control group (n = 22) (p < .05). The concentration of hippuric acid in printers was significantly higher than in the control group (p < .01), and the level of blood toluene at the end of the workshift was 0.500 mg/l. The authors also examined rotogravure printing inks-considered a potential source of genotoxic polycyclic aromatic hydrocarbons because they contained carbon black their use in printing plants, and previous documentation of increased chromosomal aberrations in rotogravure printers. Only milligrams of fluorene and phenanthrene per gram of the printing inks were found; no polycyclic aromatic hydrocarbons with carcinogenic properties were discovered in the inks. The authors used Salmonella typhimurium indicator strains TA 98, TA 100, TA 1537, and YG 1041 in spot tests and indicator strains TA 98 and TA 100 in plate-incorporation assays to determine that there was no bacterial mutagenicity of all four colors of rotogravure inks. Urinary mutagenicity, which was evaluated with a microsuspension assay containing YG 1041 indicator strain both in the presence and absence of metabolic activation, was also studied. No significant difference in bacterial mutagenicity was found between the exposed and control groups. The increased percentage of aberrant cells in printers can be explained by exposure to genotoxicants that are not excreted in urine. Toluene was the most likely cause of the aberration. PMID- 11005433 TI - Concentrations of organochlorine pesticides in milk of Nicaraguan mothers. AB - Breast-milk samples from 101 mothers from the basin of Rio Aloya, Nicaragua, were collected on two occasions within the first trimester of lactation. Milk samples were analyzed for 13 organochlorine pesticides: (1) p,p' dichlorophenyldichloroethylene; (2) p,p'-dichlorophenyltrichloroethane; (3) p,p' dichlorophenyldichlorodiene; (4) alpha-hexachlorocyclohexane; (5) beta hexachlorocyclohexane; (6) gamma-hexachlorocyclohexane; (7) delta hexachlorocyclohexane; (8) toxaphene; (9) dieldrin; (10) endrin; (11) aldrin; (12) heptachlor; and (13) heptachlor-epoxide. Organochlorines of the dichlorodiphenylethane class (i.e., p,p'-dichlorodiphenylethane and p,p' dichlorodiphenylethane) were found in all samples and at the highest mean concentrations observed in the study. Chemicals in the hexachlorocyclohexane family (i.e., alpha- and delta-hexachlocyclohexane) were not found at all (0%), and the other hexachlorocyclohexane compounds (i.e., beta > gamma) were found in less than 6% of the samples. Twenty percent or less of the sample contained chlorInated cyclodienes (i.e., dieldrin > endrin > heptachlor-epoxide > heptachlor). No measurable concentrations of alpha-hexachlorocyclohexane, aldrin, p,p'-dichlorophenyldichlorodiene, and toxaphene were found in the breast milk samples. Analysis of variance demonstrated that only the concentration of p,p' dichlorophenyldichloroethylene p,p'-dichlorophenyltrichloroethane, and endrin were affected significantly by maternal age. Overall, with the exception of p,p' chlorophenyldichloroethylene, and p,p'-dichlorophenyltrichloroethane, the mean concentrations of the analyzed pesticides were low. Total p,p' dichlorophenyltrichloroethane concentrations that exceeded the allowed daily intake set by the World Health Organisation were found in 5.9% of the samples. PMID- 11005434 TI - Effect of ventilation systems on prevalence of symptoms associated with "sick buildings" in Brazilian commercial establishments. AB - In this questionnaire study, the authors compared the prevalence of certain symptoms and signs associated with Sick Building Syndrome and perceptions relative to environmental discomfort of employees in a central-air-conditioned dropping center and in natural-ventilation commercial shops located on the streets of Niteroi, Rio de Janeiro, Brazil. There was a statistically significant higher prevalence of symptoms (e.g., water and itching of eyes, sore throat, nose irritation, difficulty breathing, skin irritation, sneezing) that were characteristic of Sick Building Syndrome in the air-conditioned building than in the naturally ventilated stores. The results indicated that there exists a continuing need for further research in Brazilian work environments. PMID- 11005435 TI - Mast cell disorder to be ruled out in MCS. PMID- 11005436 TI - Arsenic body burden and morbidity and mortality. PMID- 11005437 TI - Laparoscopic radical prostatectomy: feasibility studies or the future standard technique? PMID- 11005438 TI - Paediatric voiding dysfunction and enuresis. AB - Urinary incontinence and nocturnal enuresis are frequently encountered problems in children. In this review some aspects will be highlighted. Especially the different types of monosymptomatic nocturnal enuresis as well as some recent developments in children with dysfunctional voiding will be discussed. PMID- 11005439 TI - Non-bacterial cystitis. AB - The concept of non-bacterial cystitis (NBC) combines sterile urine and cystitic symptoms as well as inflammatory changes, in particular in the mucosa and submucosa of the bladder. It includes a multiplicity of vicious circles along the entire continence reflex. An understanding of NBC presupposes knowledge of the origin of the normal urinary urge and its successful control. Against the background of the steadily increasing incidence of interstitial cystitis (often irreversible end-stage NBC), it is suggested here that in the face of a failure of first-line therapeutics (anticholinergics, cyclic antidepressants or oestrogens), one must consider without delay the possible presence of NBC. PMID- 11005440 TI - Urethral pathology. AB - In the present review the eligible data on urethral pathologies are critically analysed, with special attention given to unresolved controversies in this topic. The entities discussed are as follows: female circumcision as a ritualistic tradition, with many associated complications; prostatourethritis and urethral syndrome; urethral diverticula as an overlooked rare pathology in urological practice; urethrorectal fistula; pelvic fracture urethral injuries and its current standard treatment algorithm; and urethral strictures and new endoscopic or surgical reconstructive techniques. PMID- 11005441 TI - Urinary diversion and reconstruction. AB - Orthotopic bladder augmentation and substitution has been established as the standard procedure for urinary diversion in many institutions, with current studies reporting mainly on continence rates and procedure-associated complications, such as the risk of impairment of neobladder function by local tumor recurrences in the small pelvis. Similarly, in other types of continent diversion, such as continent cutaneous diversion and rectal reservoirs, current interest is primarily directed towards minimization of surgery-associated complications. PMID- 11005442 TI - Defining the biologic role of genes that regulate prostate cancer metastasis. AB - Metastasis is the most lethal attribute of a cancer. There is a critical need for markers that will accurately distinguish those histologic lesions and disseminated cells that have a high probability of causing clinically important metastatic disease from those cells that will remain indolent. Despite the explosion in new information regarding the genetics of cancer, only six human genes have thus far been shown to functionally suppress metastasis. The present review and perspective describes the evolving view of the mechanisms that regulate metastasis, and the importance of metastasis-suppressor genes in this process. Specifically, the clinical problem of metastatic prostate cancer, the identification of metastatic colonization as a therapeutic target, and the identification and functional characterization of prostate cancer metastasis suppressor genes are discussed. PMID- 11005443 TI - The timing of hormonal treatment for prostate cancer. AB - This article reviews publications investigating the optimal timing of androgen ablative therapy for prostate cancer. We regard with some concern retrospective series, the use of endpoints other than overall survival, the presentation of actuarial and not actual survival, and the omission of appropriate controls. Treatment of minimal residual disease and other clinical scenarios are explored. PMID- 11005444 TI - Cryosurgery: is it an effective option for patients failing radiation? AB - Cryotherapy of the prostate represents a potential treatment for localized recurrent prostate cancer after radiation therapy. Current salvage cryotherapy of the prostate can result in undetectable serum PSA levels with low morbidity. Further refinements in technique and equipment may enhance cryosurgical results. PMID- 11005445 TI - The effects of androgen deprivation on the prostate gland: cell death mediated by vascular regression. AB - Androgenic steroids are required to maintain the prostate gland in the adult state. Consistent with this requirement, androgen deprivation therapies typically induce a drastic regression of mature prostate tissue that is accompanied by the extensive loss of prostate cells through the programmed cell death process referred to as apoptosis. Whereas, in the past, the loss of prostate cells associated with androgen deprivation has generally been perceived to be a direct response of the androgen receptor-expressing prostate cells to an androgen depleted environment, more recent studies of the prostate regression process suggest that it might instead be initiated by an indirect response of the prostatic parenchyma to an ischemic/hypoxic environment caused by a drastic reduction of blood flow to the tissue that occurs when androgens are withdrawn. This article reviews evidence that the prostatic vascular system is a primary target of androgen action and other evidence suggesting that the regression of the prostate parenchyma occurs secondarily to the regression of the prostate vascular system through cell death mediated by tissue ischemia/hypoxia. PMID- 11005446 TI - Imaging renal masses. AB - Imaging plays a crucial role in the detection, characterization, and post operative follow-up of renal masses. With rapidly advancing technology, imaging techniques are continuously evolving. This review will discuss the current modalities and techniques available for renal imaging, and recent developments. PMID- 11005447 TI - Should radical nephrectomy be performed in the face of surgically incurable disease? AB - The role of cytoreductive nephrectomy in the management of metastatic renal cancer remains controversial. Recent trials, like SWOG 8949 have suggested the usefulness of this approach at least in selected patients with good performance status and other favorable indicators. The timing of cytoreductive nephrectomy has also been controversial and remains so to this time. PMID- 11005448 TI - Immunotherapy: are we making a difference? AB - There is a 10-20 year history of immunotherapeutic approaches in metastatic renal cell cancer, which have produced a consistent demonstration of anti-tumour effect in a small percentage of patients. Clarification of dose and schedule of current agents continues to be modified. New technologies for immune system activation are attempting to enhance the response rate and improve outcome. Anti proliferative effects of immunotherapy produce prolongation of stable disease, and new agents are being developed to enhance this approach. PMID- 11005449 TI - Can biological markers predict recurrence and progression of superficial bladder cancer? AB - Biological markers that are predictive of recurrence and progression of superficial bladder tumors must provide additional information to that provided by multiplicity, size and grade. Field anomalies in normal appearing urothelium of patients with papillary superficial transitional cell carcinoma have been associated with tumor antigens and chromosome 9 deletions. Also, primary tumors with chromosome 9 deletions are associated with a higher risk of recurrence. Abnormal expression of p53, p21 and Ki-67 cell cycle markers have little predictive value for recurrence. However, p53 overexpression or mutation and decreased expression of p27 are associated with cancer progression and survival. New markers, such as mutations in the fibroblast growth factor receptor 3 gene (found in 30% of tumors), anomalies of the PTEN gene and vascular endothelial growth factor expression, may have potential and require further evaluation. Molecular fingerprints of superficial tumors with distinct clinical behavior are being rapidly unravelled. Large-scale clinical studies are urgently needed to provide supportive evidence for their incorporation in clinical management. PMID- 11005450 TI - Management of superficial bladder cancer: what is new? AB - The management of superficial bladder cancer has not changed much during the last years. Transurethral resections with adjuvant intravesical instillation to risk groups have been standard therapy. Cystoscopy and cytology have been used for follow-up. Recently combinations of drugs for instillation and new urinary markers for diagnosis and follow-up have been tested. The role of these new modalities has not been easily judged. Thus the timely advent of guidelines with policy recommendations will play an important part in improving the care of patients in the future. PMID- 11005451 TI - Advances in chemotherapy of invasive bladder cancer. AB - A survey of the literature published during 1999 identified the following advances in the chemotherapy of advanced bladder cancer. At present there are no new chemotherapy regimens superior to MTX + VBL + ADR + CDDP (M-VAC), but improvements in multidrug therapies and the development of new drugs suggest that a new first-line chemotherapy will be established. The efficacy of neoadjuvant chemotherapy could not be validated by meta-analysis, but neoadjuvant chemotherapy can be expected to be a useful tool for individualizing therapy. If it was possible to select accurately those patients who show complete response to a certain chemotherapy regimen, then this selected group would show a 100% complete response rate on that regimen, regardless of the complete response rate of this regimen in an unselected population. Such individualization of therapy would aid in future establishment of bladder-preserving treatment methods for invasive bladder cancers. PMID- 11005452 TI - Outcome of radical cystectomy for invasive bladder cancer. AB - Even though radical cystectomy still remains the 'gold standard' for the treatment of invasive bladder cancer, newer insights and developments are entering the urological arena: a 'tailored' surgical approach combining a less extensive procedure and a better quality of life seems feasible for selected patients without compromising the outcome; the type of urinary diversion has no impact on the risk of complications, the ability to receive postoperative salvage treatments and the natural history of the disease; the depth of extension of the tumour and the nodal involvement are the only independent 'classical' predictors of survival after radical cystectomy; pelvic node dissection is curative in patients with limited nodal involvement; the clinical application of newer molecular prognostic factors still remains controversial. PMID- 11005453 TI - Prognostic parameters for the management of advanced testis tumours. AB - The need for prognostic parameters in testicular germ cell tumours is sometimes questioned based on an overall cure rate of more than 80% of the patients regardless of tumour stage. However, the trend for an earlier and more accurate diagnosis amenable to curative treatment as well as the high effectiveness of standard Cisplatinum containing chemotherapy has masked the continuing need for intensifying therapy in patients with adverse risk factors. This intense treatment is often associated with worrysome morbidity and the assessment of prognostic factors, stage by stage, is warranted on which patient at risk can be identified and treated accordingly. Traditional prognostic factors, on which most classification systems are based, include large tumour volume, the presence of liver, bone or brain metastasis, grossly elevated tumour markers and an extragonadal primary site, particularly in the mediastinum. Novel prognostic factors are either (1) independent from the patient and his disease, (2) inherent on the patient's characteristics or (3) based on tumour biology. Clearly, the infrastructure and the experience of the treating uro-oncology unit (see 1) is decisive for treatment outcomes, and -at least-'difficult to treat' patients should be referred to properly resourced cancer centres. Patients with higher socio-economic status, willing to travel and well educated enough to be worried about their diseases status apparently gain access to expert centres more easily (see 2), translating into an upgrade on prognosis. Finally, biologic factors (see 3) such as beta-human chorionic gonadotrophin or MAGE epitopes in seminoma or the percentage of embryonal carcinoma components orvascular invasion mayor may not inversely influence the prognosis and need further assessment in prospective trials. However, the search for even better (molecular) biologic factors is speeding up because more complex treatment decisions such as in advanced testicular cancers rely on a more precise determination of prognosis, enabling a more tailored selection of individualized therapeutic options. PMID- 11005454 TI - Sexual function and fertility after treatment of testicular cancer. AB - As a result of the introduction of effective cisplatin-based chemotherapeutic regimens into the clinical routine, even patients with metastatic testicular cancer at initial diagnosis can be cured of their disease. Sexual dysfunction and infertility are common long-lasting sequelae in testicular cancer survivors, affecting approximately 20% of patients after the application of the different treatment modalities currently available for the treatment of early and advanced clinical stages, including retroperitoneal surgery and systemic chemotherapy. Accordingly, it has been demonstrated that fertility distress and sexual disturbances, the latter occurring in only a minority of patients after surgical or chemotherapeutic treatment of testicular germ cell tumours, substantially alter the patients' quality of life. It is even worse because testicular cancer mostly affects men in the prime of their physical, sexual and reproductive function. Although semen quality is frequently poor at initial diagnosis and further deteriorates after orchiectomy, probably because of structural abnormalities in the remaining contralateral testicle, the advent of intracytoplasmatic sperm injection promises a fertile future to most patients, even if only a few sperms are present in the ejaculate. Further long-term investigations should be initiated to clarify the impact of the different treatment modalities on fertility and sexual life. The main objective should be the identification of patients who are at increased risk of developing therapy related physical and psychological problems. PMID- 11005455 TI - Bibliography. Current world literature. Voiding dysfunction and female urology. PMID- 11005456 TI - Bibliography. Current world literature. Oncology: prostate and renal. PMID- 11005457 TI - Bibliography. Current world literature. Oncology: bladder and testis. PMID- 11005458 TI - Overview: pulmonary thromboembolism. PMID- 11005459 TI - Incidence of pulmonary embolism: is it declining? AB - Several factors make analysis of time trends in the incidence of pulmonary embolism difficult. Apart from the methodologic problems, a number of factors could explain an increasing as well as a decreasing incidence. The few studies on pulmonary embolism discovered after death are discussed, and the evidence shows a decrease in incidence, especially after the mid 1970s. This can be explained by many factors, of which the 2 most important are the increased use of thromboprophylaxis and better treatment of established deep vein thrombosis. PMID- 11005460 TI - Current status of pulmonary embolism and venous thrombosis prophylaxis. AB - Prophylaxis for venous thromboembolism is an area that has received intense study in certain conditions, but less than adequate coverage in other areas. In considering who needs prophylaxis, patients are categorized into levels of risk. Clinical venous thromboembolism can be correlated to these levels of risk. Methods of prophylaxis include pharmacologic, mechanical, and combinations of these. Each category of surgical and medical patient requires specific types of prophylaxis. In certain orthopedic indications, the length of prophylaxis outlasts the inpatient hospital stay and may be as long as 1 month after discharge. Even with the best prophylaxis today, the incidence of deep venous thrombosis (DVT) and pulmonary embolism (PE) is decreased by only approximately 70% to 80%. Further developments should allow for greater declines in the rates of venous thromboembolism, with its subsequent short-term consequence of pulmonary embolism and lower extremity morbidity and long-term consequence of the disabling syndrome of chronic venous insufficiency (CVI). PMID- 11005461 TI - Current diagnostic techniques for pulmonary embolism. AB - The diagnosis of venous thromboembolism (VTE) includes deep venous thrombosis (DVT) and pulmonary embolism (PE) and requires objective testing. The clinician uses a combination of risk factors and nonspecific clinical findings to identify patients who warrant such an evaluation. The recommended approach begins with ventilation/perfusion (V/Q) lung scans or lower extremity noninvasive studies by compression ultrasonography. Nondiagnostic V/Q scans or negative noninvasive studies require further testing. A high-probability V/Q scan or a positive noninvasive study warrant treatment. A normal V/Q scan excludes the diagnosis of PE. Helical computed tomography (CT) can diagnose PE of major vessels but is not sufficiently sensitive to exclude PE because of its poor sensitivity for subsegmental pulmonary vessels. Newer D-dimer assays have a high negative predictive value, but results vary with the specific assay and do not perform well in patients with cancer. Future studies are needed to validate magnetic resonance imaging (MRI) or magnetic resonance angiography (MRA). PMID- 11005462 TI - Low-molecular-weight heparin in the treatment of pulmonary embolism. AB - Unfractionated heparin is the traditional treatment of choice for patients presenting with submassive pulmonary embolism. However, newer low-molecular weight heparins have been developed and tested in several clinical trials in the last 2 decades because of the many limitations of unfractionated heparin. The pharmacologic properties of the low-molecular-weight heparins allow them to be administered subcutaneously in fixed, weight-adjusted doses without need for laboratory monitoring. Different studies have recently proved their efficacy in the treatment of patients with deep vein thrombosis. Because there is good evidence that deep vein thrombosis and pulmonary embolism are 2 expressions of a single disease, the use of low-molecular-weight heparins has been extended to patients presenting with clinically stable pulmonary embolism. In particular, the results of 2 important clinical trials, the Columbus and the Thesee, have conclusively proved that the low-molecular-weight heparins are as safe and effective as unfractionated heparin. In some centers, approximately 80% of patients with deep vein thrombosis are treated at home. This suggests that carefully selected patients with clinically stable pulmonary embolism can receive home treatment with low-molecular-weight heparins. PMID- 11005463 TI - Indications for filter placement. AB - Indications for placement of a vena caval filter reflect the perception of efficacy and safety of these devices. The traditional indications include contraindications to or complications of anticoagulation. Over time, physicians have expanded these indications to include prophylactic placement in patients at high risk of pulmonary embolism. The decision to place a filter should be made only after weighing the risks and benefit in each situation. Long-term outcomes should be carefully evaluated. PMID- 11005464 TI - Filter placement by ultrasound technique at the bedside. AB - Vena cava filtration devices are commonly inserted for the prevention of fatal pulmonary emboli. Several imaging methods are used today to direct device placement. Bedside insertion has several advantages, particularly in the critically ill patient. Imaging techniques include bedside fluoroscopy, transperitoneal ultrasound, and intravascular ultrasound. We have now successfully placed 10 transfemoral vena cava filters by use of intravascular ultrasound at the patient's bedside. This appears to be a feasible method for filter insertion. PMID- 11005465 TI - Current vena cava filter devices and results. AB - The critical focus in the acute management of deep vein thrombosis (DVT) continues to be the effective prevention of associated life-threatening pulmonary embolism (PE) and the achievement of survival for those in whom such emboli occur. As such, primary treatment for DVT or PE demands adequate anticoagulation with intravenous heparin followed by maintenance oral warfarin for 3 to 6 months. This centered approach successfully prevents 90% to 95% of cases of associated pulmonary embolism. However, under circumstances in which anticoagulation cannot be used, or in which it fails, there is clear indication for placement of a mechanical filter into the inferior vena cava to provide a protective barrier against the passage of clinically significant emboli from pelvic or lower extremity veins. Considerable technical ingenuity and continued evolution of materials and design have propelled the development and number of vena cava filters available for clinical use. Variable data on safety and effectiveness require physicians to match the best filter to each patient's particular situation and anatomy. However, many interventional radiologists and surgeons often base their filter selection on ease of insertion and device cost. Thus, it is paramount that physician users of these devices remember that the primary objective of vena cava filtration is to provide a safe and effective device for permanent implantation. If this objective is not met, the quality of care in the management of deep venous thrombosis or pulmonary embolism will diminish. PMID- 11005466 TI - Filter complications and their management. AB - Complications from the use of vena caval filters are rare but can occur at placement, during other procedures, or late in follow-up. Placement problems such as bleeding from the insertion site or embolism from thrombus manipulation are largely avoidable. Technical difficulties with introduction, positioning, or misplacement have been minimized by use of flexible carriers and guidewires. However, later insertion of guidewires without fluoroscopy for line changes can lead to problems of entrapment. Filter capture of emboli can result in further thrombus propagation at that site or in filter obstruction. Management of these events requires adequate visualization to determine whether to use lytic therapy or a second filter. Late complications include filter strut fracture in 0.05% of cases and filter penetration, of even rarer functional significance. Pain that can be attributed to the filter has been found only in a single case of pelvic misplacement. The limited consequences of mechanical changes in the filter argue strongly against unnecessary interventions, which frequently result in an increase in morbidity. PMID- 11005467 TI - The current role of thrombolytic therapy for pulmonary embolism. AB - Thrombolytic therapy for pulmonary embolism can rapidly reverse right ventricular failure and reduce mortality and morbidity among appropriately selected patients. Individuals being considered for this treatment should be screened for potential major bleeding problems, which, if present, should lead to alternative management with catheter or surgical embolectomy. There is no ideal thrombolytic agent; nor have indications for thrombolysis been precisely defined. Available data indicate that patients with moderate or severe right ventricular dysfunction gain the most from this pharmacologic strategy. PMID- 11005468 TI - Catheter technique for pulmonary embolectomy or thrombofragmentation. AB - The management of acute massive pulmonary embolism (PE) constitutes a major clinical problem because of the associated derangement of hemodynamic and respiratory functions from obstruction to pulmonary blood flow. Despite advances in management with thrombolytic therapy or open embolectomy, the mortality rate remains high. To improve the chance of survival, catheter techniques that are capable of removing or fragmenting the clot have been developed. These include catheter pulmonary embolectomy and thrombofragmentation. The former involves the introduction of a suction catheter from a femoral or jugular venotomy through the right heart into the appropriate pulmonary artery under fluoroscopic guidance. The technique for the latter involves the percutaneous introduction of a fragmentation catheter from a femoral vein through a guiding catheter into the appropriate pulmonary artery. The success of the catheter technique in removing pulmonary emboli varies with different devices. The overall success rate is approximately 76%, with a mortality rate of 25%. Transvenous pulmonary embolectomy and thrombofragmentation are safe and effective techniques for treating patients with massive PE. The success of each of the techniques depends on a thorough understanding of the mechanism of action of each of the devices used and a facile catheterization technique. PMID- 11005469 TI - Indications for and the results of pulmonary thromboendarterectomy for thromboembolic pulmonary hypertension. AB - Most episodes of acute pulmonary embolism completely resolve, through thrombolytic mechanisms. When adequate thrombolysis fails, however, residual obstruction to pulmonary vascular flow by fibrotic remnants may result in pulmonary hypertension. The genesis of this pulmonary hypertension is complex. Apart from the simple mechanical obstruction to blood flow that may occur from occlusion of significant areas of the pulmonary vascular bed, secondary vascular changes eventually may result from increased flow and pressure in nonobstructed areas of the lung, or factors generated by localized hypoxia in occluded segments. Chronic pulmonary hypertension from thromboembolism, regardless of the mechanism, is much more prevalent than is generally appreciated. Patients develop increasing shortness of breath, and the diagnosis is often missed. Pulmonary endarterectomy offers a cure for the condition, and wider recognition of the efficacy of the operation and the entity are therefore important. Pulmonary endarterectomy is a technically demanding procedure, now performed with success at only a handful of centers. However, with proper patient selection, meticulous surgical technique, and careful postoperative management, excellent results can be obtained. A true endarterectomy (not an embolectomy) of all affected parts of the lung is performed, and cardiopulmonary bypass, systemic cooling, and circulatory arrest are essential for the visibility necessary to clear all affected areas of the pulmonary vasculature. Medical treatment is ineffective for this condition, and the alternative surgical treatment is lung transplantation. Pulmonary endarterectomy appears to be permanently curative, though an inferior vena caval filter should be placed in all patients to prevent recurrence, and the patients must have life-long anticoagulation. PMID- 11005470 TI - Transvaginal approach to Cooper's ligament during incontinence surgery. PMID- 11005471 TI - Intraurethral ultrasonography: correlation of urethral anatomy with functional urodynamic parameters in stress incontinent women. AB - To determine whether differences in functional urodynamic parameters can be explained by changes in urethral anatomy, 39 patients underwent intraurethral ultrasonography to obtain a 360 degree view of the urethra. The point of maximal rhabdosphincter thickness was identified in all patients. The thickness, circumference and area of the urethral smooth and skeletal muscle layers were calculated. Data from patient histories and urodynamic evaluations were compared with this anatomical survey. The urodynamic diagnoses were as follows: 10 patients were normal, 24 had genuine stress incontinence and 5 had intrinsic sphincter deficiency. These patients had decreasing rhabdosphincter thicknesses of 3.91, 3.35 and 2.70 mm (P = 0.048). A weak linear relationship was found between maximal urethral closure pressure and rhabdosphincter (r = 0.40, P = 0.013) and longitudinal smooth muscle (r = 0.35, P=0.027) thickness. It was concluded that a loss of urethral resistance as measured by maximal urethral closure pressure is associated with changes in urethral anatomy identified by intraurethral ultrasonography. PMID- 11005472 TI - Pelvic muscle exercise effect on pelvic muscle performance in women. AB - The aim of this study was to compare pelvic muscle (PM) characteristics (strength, endurance and contractability) before and after 12 weeks of pelvic muscle exercises in two groups of older women: the first composed of women with genuine stress incontinence, and the second made up of women with no symptoms of urinary incontinence or pelvic organ prolapse. This research also investigated the extent to which PM pressure and health-related characteristics could help discriminate between women with and without a clinical sign of PM dysfunction. Within a framework of skeletal muscle fitness, outcome measures were defined and compared. There was no significant difference in the baseline (P = 0.09) and post PME (P = 0.63) strength, endurance and contractability of the two groups of women. The two groups did differ significantly on change scores (P = 0.05) following PME. A greater improvement in strength for women without a clinical sign of dysfunction was demonstrated. There was a probability of 91% that those with a history of gynecological surgery belonged to the group of women with SUI. PMID- 11005473 TI - Bacterial vaginosis increases in pessary users. AB - The purpose of this study was to examine the association between pessary use, smoking and changes in the vaginal flora. Patients using pessaries were age matched with non-pessary using controls. All candidates examined were women attending the Mount Sinai Hospital, Toronto, for genitourinary problems. Vaginal cultures were routinely performed on all women attending the unit, irrespective of symptoms. Forty-four pessary users were age matched with 176 controls (4 controls per case). The mean age was 60.1 +/- 12.6 years, and 15% of these were premenopausal. The duration of pessary use ranged from 0.5 to 8 years (mean 3.3 +/- 1.7). Weight, parity, smoking status, diabetes mellitus, thyroid disease, UTI and postvoid residual urine volume were not significantly different between pessary users and controls. Bacterial vaginosis (BV) was noted in 32% of pessary users, versus 10% of controls. The relative risk of developing BV in pessary users was 3.3 (OR, 4.37; 95% CI, 2.15-9.32), P = 0.0002. Smoking independently affected the vaginal flora, increasing the relative risk of developing BV to 2.9 (OR, 3.78; 95% CI, 2.05-8.25), P = 0.0013. It was concluded that pessary use is a very effective and conservative method for the treatment of genital prolapse. However, we found that the presence of a foreign body was associated with changes in the vaginal flora, thereby increasing the odds of developing bacterial vaginosis to 4.37; this was further compounded by smoking. PMID- 11005474 TI - Orthotopic ileal neobladder in females: impact of the urethral resection line on functional results. AB - We present our functional experience with orthotopic bladder replacement in female patients dependent on the urethral resection line. Between November 1986 and July 1998 42 women underwent orthotopic urinary tract reconstruction with an ileal neobladder at our institution: 26 patients underwent radical cystectomy (RCx) with subsequent ileal anastomosis to the urethra, and 16 underwent simple cystectomy (SCx) with preservation of the bladder neck. Fourteen of 22 patients following RCx and 3 of 14 patients following bladder neck-sparing cystectomy void naturally. Clean intermittent catheterization is necessary in 8 of 22 and 11 of 14 patients, respectively. Perfect continence with no pads at 1 year postoperatively was achieved in 15 of 18 evaluable patients following RCx and 10 of 11 patients following bladder neck sparing. Incontinence requiring one or more pads is present in 3 of 18 patients and 1 of 11 patients, respectively. Subjectively satisfactory continence was achieved in 16 of 18 patients following RCx and in all patients following bladder neck-sparing surgery. Our conclusions are that radical cystectomy as well as a bladder neck-sparing cystectomy does provide satisfactory functional results in the majority of patients. However, the urethral resection line does slightly influence the rate of incontinence as well as the requirement for intermittent catheterization. RCx does translate into a lesser requirement for CIC, whereas bladder neck sparing results in slightly better continence rates. PMID- 11005475 TI - Patient satisfaction and complications following sacral nerve stimulation for urinary retention, urge incontinence and perineal pain: a multicenter evaluation. AB - The aim of the study was to determine the success rate, the complications, the failures and the solutions found in troublesome cases. A retrospective study was performed in three university centers in Belgium. Between March 1994 and April 1998, a quadripolar electrode and a pulse generator were implanted in 53 patients (8 men, 45 women, 43 +/- 12 years, mean follow-up 24 +/- 8 months, range 13-39 months). During the first few months, 45 (85%) of the 53 patients had an objective response. Eight late failures occurred, with a mean failure delay of 9 +/- 5 months. We performed 15 revisions in 12 patients. Major complications were pain and current-related troubles. The outcome was significantly better (P = 0.001) in post-stress incontinence surgery patients. Device-related pain was found more frequently in patients with dysuria and/or retention or perineal pain, and the test stimulation was less reliable (P = 0.025) in patients with a psychiatric history. Sacral nerve stimulation is efficient in treating patients with refractory lower urinary tract symptoms and/or perineal pain. PMID- 11005476 TI - 'Nerve-sparing' cystectomy in women. AB - Although the indications for cystectomy other than for cancer are few, it is occasionally necessary for severe interstitial cystitis or hemorrhage due to radiation cystitis. The use of substitution cystoplasty after cystectomy has increased in men in the last decade, and this has resulted in the development of 'nerve-sparing' approach to cystectomy to improve continence and potency. The use of substitution cystoplasty in women after cystectomy has lagged behind that in men because it was considered necessary to remove the entire urethra, making orthotopic substitution impossible. Recently the need to remove the entire urethra has been questioned, with the finding that if the bladder neck is free of tumour, recurrence of disease in the urethra has not so far been found to occur. In addition, it appears possible to substitute on to the urethra without incontinence. This has led to the development of a 'nerve-sparing' technique of cystectomy, and renewed interest in the anatomy of the urethra. This paper reviews the current literature on 'nerve-sparing' cystectomy and describes our technique. PMID- 11005477 TI - Posthysterectomy vault prolapse. AB - The incidence of vault prolapse is uncertain but appears to be increased five fold after vaginal hysterectomy. A precise scoring system has now been devised to overcome the diagnosis and classification of this condition. Conservative measures may be used to treat this condition in women unfit for surgery or those who require symptomatic relief while awaiting surgery. The surgical options lie between a vaginal sacrospinous fixation, or abdominal procedures such as sacrocolpopexy or vault suspension operations. The success rate for these operations is over 90%. Pre-operative urodynamic evaluation is mandatory since these patients frequently have concomitant stress urinary incontinence which may require correction at the same time. The choice of operation will undoubtedly depend upon the experience of the surgeon but greater awareness and alteration of technique at the time of the original hysterectomy may be the better solution to reducing the incidence of vault prolapse. PMID- 11005478 TI - Bladder substitution in women. AB - Although numbers are still small and follow-up is often limited, female orthotopic bladder reconstruction has been shown to provide excellent, near normal voiding and storage function in selected patients, and its popularity has been growing over the last decade. The true role of this form of surgery, however, is not yet fully established. This paper discusses the indications for patient selection, operative technique and outcome of female bladder substitution. Outcome is equivalent to that seen in male patients, although in some series there is a higher rate of voiding dysfunction requiring intermittent self-catheterization. Although further data regarding urethral recurrence rates are awaited, it would seem that with careful patient selection and operative technique, cancer surgery may not be compromised. Functional results have been excellent and patient satisfaction is very high. Provided there is a functional external sphincter and tumor margins can be safely cleared, this form of surgery offers patients the best opportunity to preserve quality of life following cystectomy. PMID- 11005479 TI - Urodynamic assessment of voiding dysfunction and dysfunctional voiding in girls and women. AB - Voiding dysfunction is defined as impaired bladder emptying, and presents with a mixture of lower urinary tract symptoms. Dysfunctional voiding is a condition in which there is a lack of coordination between the sphincter and detrusor during emptying in a patient without overt uropathy or neuropathy. Assessment of voiding dysfunction is important in women and girls in the prevention and treatment of urinary incontinence, retention, urinary tract infection and subsequent kidney damage. Accurate diagnosis is essential in order to select the correct treatment. Screening can be done by history-taking: symptom scores can help to guide the screening. More objective measures are uroflowmetry, ultrasonography and video urodynamics. The latter is the gold standard for the diagnosis of voiding dysfunction and consists of simultaneous registration of pressure in the bladder and rectum and external sphincter behavior, either by electromyographic recording of pelvic floor activity or by pressure recording at the external sphincter, during the whole bladder cycle of filling and emptying. On fluoroscopy the bladder can be visualized throughout the filling and emptying phase. In dysfunctional voiding, hypertonicity and instability of the external urethral sphincter during filling cystometry and impaired external sphincter relaxation during emptying are pathognomonic findings. Pressure-flow analysis reveals no obstruction and the detrusor contractility is low. PMID- 11005480 TI - The use of mesh in gynecologic surgery. PMID- 11005481 TI - Practice parameters for antibiotic prophylaxis to prevent infective endocarditis or infected prosthesis during colon and rectal endoscopy. The Standards Task Force. The American Society of Colon and Rectal Surgeons. PMID- 11005483 TI - A bold message for the next millennium. PMID- 11005482 TI - Practice parameters for antibiotic prophylaxis--supporting documentation. The Standards Task Force. The American Society of Colon and Rectal Surgeons. PMID- 11005484 TI - Patient satisfaction after surgical treatment for fistula-in-ano. AB - PURPOSE: The surgical treatment of fistula-in-ano frequently results in recurrence of the fistula or postoperative anal incontinence. Despite these problems, most patients are satisfied with the results of their surgery. To clarify this apparent discrepancy, we attempted to identify factors that affect patient's lifestyles and may contribute to their satisfaction. METHODS: A questionnaire was mailed to 624 patients surgically treated for cryptoglandular fistula-in-ano at the University of Minnesota during a five-year period. Three hundred seventy-five patients returned their questionnaires. Patients who were followed up for a minimum of one year were included in this retrospective study. Associations between postoperative complications and patient satisfaction were identified by chi-squared tests and multiple logistic regression. Attributable fractions for patient dissatisfaction were calculated using study population dissatisfaction rates. RESULTS: Patient satisfaction was strongly associated with fistula recurrence, difficulty holding gas, soiling of undergarment, and accidental bowel movements. Effects of incontinence on patient quality of life were also significantly associated with patient satisfaction as was the number of lifestyle activities affected by incontinence. Patients with fistula recurrence reported a higher dissatisfaction rate (61 percent) than did patients with anal incontinence (24 percent), but the attributable fraction of dissatisfaction for incontinence (84 percent) was greater than that for fistula recurrence (33 percent). Patient satisfaction was not significantly associated with age, gender, history of previous fistula surgery, type of fistula, surgical procedure, time since surgery, or operating surgeon. CONCLUSION: Patient satisfaction after surgical treatment for fistula-in-ano is associated with recurrence of the fistula, the development of anal incontinence, and with the effects of anal incontinence on patient lifestyle. In our series of patients treated mainly with laying open of the fistula tract, patients with fistula recurrence had a higher dissatisfaction rate than did patients with anal incontinence. However, because anal incontinence was more prevalent than fistula recurrence, a higher fraction of dissatisfaction was attributable to anal incontinence. PMID- 11005485 TI - Restoring control: the Acticon Neosphincter artificial bowel sphincter in the treatment of anal incontinence. AB - PURPOSE: Anal incontinence is a socially disabling problem affecting 1 to 2 percent of the population. Anal sphincter replacement is a treatment option if the problem is severe and not amenable to direct repair. The artificial bowel sphincter is an innovative approach. We report the technique for placement and the outcomes which have occurred in an initial series of 13 patients. METHODS: The Acticon Neosphincter artificial bowel sphincter consists of an inflatable cuff of silicone elastomer placed around the anal canal and connected to a pressure-regulating balloon in the iliac fossa via a control pump placed in the labium or scrotum. Thirteen patients with severe anal incontinence not amenable to other methods were treated. Causes of incontinence included obstetric damage in eight patients, surgical damage in two patients, imperforate anus in two patients, and spina bifida in one patient. RESULTS: Surgical placement of the device was straightforward, mean operating time was 65 minutes, and median length of stay was 3.6 days. One infection of the perineal wound occurred in the early postoperative period necessitating removal of the device. In two further patients the artificial bowel sphincter was removed because of late infection in one at seven months and because of erosion through the skin in another at three months. The artificial bowel sphincter has been activated in ten patients resulting in full continence to solids and liquids except in one patient with postvagotomy diarrhea who had some leakage of liquids during episodes of diarrhea. The mean (+/- standard deviation) continence score (Cleveland Clinic system; maximal incontinence = 20) changed from 18.7 +/- 1.6 preoperatively to 2.1 +/- 2.6 after activation (P < 0.0001). Quality of life measured using a continence-specific series of up to 39 questions changed from 77 +/- 16 percent of maximal reduction of quality preoperatively to 12 +/- 19 percent postoperatively (P < 0.001). CONCLUSIONS: The artificial bowel sphincter can be placed without technical difficulty and with low morbidity. Preliminary experience shows full restoration of continence in most patients and ease of use. Longer follow-up is needed to determine the extent of problems with infection, erosion, and mechanical failure. PMID- 11005486 TI - Outcome of patients with rectal adenocarcinoma and localized pelvic non-nodal metastatic foci. AB - PURPOSE: The aim of this study was to evaluate the outcome of patients with primary rectal adenocarcinoma and soft tissue metastatic foci restricted to the pelvis and to determine whether this entity, which is considered N1 disease in the American Joint Committee on Cancer staging system, behaves like completely replaced nodal disease or the first sign of M1 disease. The clinical course for patients with this finding is not well-described in the literature. METHODS: The authors retrospectively reviewed the medical records of 395 patients with rectal adenocarcinoma who received radiation treatment. Eighteen patients had pelvic soft tissue metastatic foci. Exclusions from this study included 1) cases without metastatic pelvic foci; 2) cases of recurrent cancer; 3) cases with known distant metastatic disease as defined by American Joint Committee on Cancer criteria; and 4) cases with extrapelvic metastatic foci. All patients received adjuvant radiotherapy. Thirteen cases received preoperative radiotherapy. Four cases received postoperative radiotherapy. One case received both preoperative and postoperative radiotherapy. Eight cases received chemotherapy. RESULTS: All eighteen patients had T3 or T4 lesions. Thirteen patients had lymph nodes that contained metastatic disease and would therefore have been scored N1 or N2 even without the pelvic tumor implants. Sixteen of 18 (89 percent) patients died of disease after a survival time of 12 to 37 (mean, 25) months. Only 1 of 18 (6 percent) patients was disease free at five years. The other remaining survivor was undergoing palliative therapy for metastatic disease to the lung. This is significantly worse than our institution's experience with T3,4N+ disease after preoperative radiation (5-year survival, 11 vs. 56 percent; P = 0.0002, Generalized Wilcoxon of Breslow). There was a high incidence of local (9/18) and distant (14/18) failure. No other factor, including radiation dose, margin status, chemotherapy, T stage, and number of involved nodes or soft tissue implants, correlated independently with outcome. CONCLUSIONS: Pelvic metastatic foci confer a significantly worse prognosis than other T3,4N+ disease. Such cases should be excluded from prospective trials for localized disease. Although this entity probably represents M1 disease for most patients, survival can be long, and aggressive locoregional and systemic treatment is warranted. PMID- 11005487 TI - Colorectal cancer before the age of 40: a case-control study. AB - PURPOSE: Colorectal adenocarcinoma before the age of 40 is uncommon, and its prognosis is controversial, with many studies reporting a worse prognosis than in older patients and others showing no difference. The current study compared two groups of patients who had surgical resection for colorectal adenocarcinoma. METHODS: The case group was composed of 34 patients younger than 40 (34 +/- 4) years. Detailed pathologic prognosis factors, tumor cell proliferation measured by proliferating cell nuclear antigen, survival, family history, and predisposing conditions were analyzed. Results were compared with a control group constituted of 34 patients older than 65 (75 +/- 6) years matched by gender, cancer site, and Dukes stage. RESULTS: Tumor differentiation, presence of vascular and perineural neoplastic invasion, tumor growth pattern, tumor cell proliferation measured by proliferating cell nuclear antigen count, and survival according to the Kaplan Meier method were not significantly different between younger and older patients. The only difference between the two groups was a higher prevalence of family history and predisposing conditions for colorectal cancer in younger patients (23 vs. 3 percent; P = 0.03). CONCLUSION: This case-control study documents that pathologic features and prognosis of colorectal adenocarcinoma are comparable in patients younger than 40 years compared with older patients for identical stages. The higher prevalence of positive family history in younger patients suggests a different genetic background compared with older patients. PMID- 11005488 TI - Apoptosis in rectal cancer: prognostic significance in comparison with clinical histopathologic, and immunohistochemical variables. AB - PURPOSE: The aim of this study was to evaluate the prognostic value of the apoptotic index for recurrence and disease-free survival after curative surgery for rectal cancer, particularly in relation to clinicopathologic variables, p53- and bcl-2 expression. METHODS: Formalin-fixed, paraffin-embedded tissue samples of rectal carcinomas resected curatively within a five-year period were used (N = 160). Apoptotic cells with fragmented DNA were detected by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphatase-biotin nick-end labeling method. The ratio of apoptotic tumor cells (in percent) was classified into low apoptotic index (less than 10 percent) and high apoptotic index (10 percent or more). Immunohistochemical analysis was performed using monoclonal antibodies (DO-1 for p53 and clone 124 for bcl-2). Statistics included univariate and multivariate analysis, and survival was calculated using the Kaplan-Meier method. RESULTS: Seventy-five percent of tumors showed a low apoptotic index, and 25 percent had a high apoptotic index. No correlation was found between apoptotic index and International Union Against Cancer stage (P > 0.05). However, significant correlations were documented with histologic differentiation (mean apoptotic index, 5.74 percent in moderately vs. 3.98 percent in poorly differentiated carcinomas; P = 0.0173), lymph node involvement (mean apoptotic index, 6.11 percent in pN1 vs. 3.72 percent in pN2; P = 0.0074), p53 status (mean apoptotic index, 6.26 percent in p53- vs. 4.42 percent in p53+; P = 0.0085), and bcl-2 expression (mean apoptotic index, 5.13 percent in bcl-2- vs. 6.51 percent in bcl-2+; P = 0.0418). Tumors of the lower rectum had a lower apoptotic index than those of the upper rectum (P = 0.0277). Neither univariate nor multivariate analysis assessed apoptotic index as predictor of prognosis: Recurrence rates did not differ between tumors related to apoptotic index (22 percent with low apoptotic index vs. 15 percent with high apoptotic index; P > 0.05), and no significant differences were found regarding survival (P > 0.05). On multivariate analysis, International Union Against Cancer stage (P = 0.0002), p53 (P = 0.0002), gender (P = 0.0136), and bcl-2 (P = 0.0243) were independent predictors of recurrence. These variables, except for bcl-2, were also independently related to disease-free survival. CONCLUSIONS: Reflecting tumor biology, apoptotic index as single variable showed no prognostic significance, whereas p53 was an independent predictor for both recurrence and survival, and bcl-2 was independently related to recurrence, but not to survival. Clinically, International Union Against Cancer stage and gender were independent prognostic factors after curative surgery for rectal cancer. PMID- 11005489 TI - Selective total mesorectal excision for rectal cancer. AB - PURPOSE: Total mesorectal excision has been advocated for rectal cancer, but its use in upper rectal and rectosigmoid tumors remains a point of debate. METHODS: One hundred seventeen patients with rectal cancers were subjected to a prospective policy of total mesorectal excision for mid and low rectal cancers and a wide (5 cm) distal margin mesorectal excision for upper rectal and rectosigmoid cancers. RESULTS: Forty-one patients underwent ultralow anterior resection, 10 underwent abdominoperineal excision, 64 had anterior resection and 2 had Hartmann's procedure. The median follow-up was 39 months. Forty-three patients had a defunctioning ileostomy. Three patients (7.3 percent) had anastomotic leaks after ultralow anterior resection with total mesorectal excision. Ninety-three patients had palliative resections. There were four locoregional recurrences in this group, giving an actuarial locoregional recurrence rate of 9.3 percent at five years. The actuarial locoregional recurrence rate after anterior resection was 6.5 percent at five years. The actuarial five-year cancer-specific survival rate was 81.4 percent at five years. CONCLUSION: These results demonstrate that a policy of wide excision of the mesorectum for upper rectal and rectosigmoid cancer and total mesorectal excision for mid and low rectal cancer is associated with a low locoregional recurrence rate and may be as efficacious as routine total mesorectal excision for all rectal cancers. PMID- 11005490 TI - Does technique of anastomosis play any role in developing late perianal abscess or fistula? AB - PURPOSE: This study examines the risk factors for developing perianal abscess or fistula formation after ileal pouch-anal anastomosis procedure for chronic ulcerative colitis or familial adenomatous polyposis. METHODS: A total of 1,457 patients with J-pouch, 1,304 (89.5 percent) with chronic ulcerative colitis and 153 (10.5 percent) with familial adenomatous polyposis who had a two-stage procedure without any evidence of previous perianal disease were included in the study. The effect of pouch-to-anal anastomosis type on perianal abscess or fistula formation was evaluated. RESULTS: A total of 108 patients (7.4 percent) had a perianal abscess or fistula after the ileal pouch-anal anastomosis procedure after at least one year of follow-up. No statistically significant difference was identified in fistula formation regarding the age and gender of the patients (P > 0.05), nor did the risk of fistula formation differ significantly between the patients with handsewn vs. stapled anastomoses (P > 0.05). However, patients with a diagnosis of chronic ulcerative colitis, compared with patients with familial adenomatous polyposis, had a statistically higher risk of developing abscess or fistula (P = 0.012). CONCLUSION: The most important risk factor in developing perianal sepsis in long-term patients with ileal pouch anal anastomosis is the initial disease type. After excluding patients without Crohn's disease, the risk of developing an abscess or fistula was found to be significantly greater in patients with chronic ulcerative colitis compared with patients with familial adenomatous polyposis, and this risk is independent of anastomotic technique. PMID- 11005491 TI - Laparoscopic-assisted colonoscopic polypectomy: the Texas Endosurgery Institute experience. AB - PURPOSE: The advent of laparoscopic surgery has altered the manner by which surgical specialties address pathologies of the abdominal cavity. This advance in technology has also changed colorectal surgery. One of the more common procedures of colorectal surgery is segmental resection for polyps that are large, broad based, or inaccessible for colonoscopic removal. We present a technique combining colonoscopy and laparoscopy to remove troublesome polyps without the need for segmental resections. METHODS: From May 1990 to September 1999 laparoscopic monitored colonic polypectomies were performed in 47 patients, with a total of 60 polyps being removed. After laparoscopic mobilization of the involved segment of the colon, the proximal bowel is cross-clamped and the colonoscope passed to the involved portion of the colon. The polyp is then presented to the colonoscopist by the laparoscopist facilitating removal. The serosal surface is monitored for any indications of transluminal injury, and the area is repaired if needed. All polyps undergo immediate frozen section analysis. If the pathologic evaluation indicates malignancy then a segmental resection may be performed, otherwise the patients are decompressed and fed within a short time before discharge. RESULTS: The polyps were located most commonly in the ascending colon (18 polyps), transverse colon (12 polyps), and cecum (12 polyps). The most common histopathologic diagnosis was tubulovillous adenoma in 28 polyps followed by villous adenoma in 11 polyps. In three cases histopathologic diagnosis revealed malignancy necessitating segmental resection (1 low anterior resection and 2 right hemicolectomies), which were performed laparoscopically. Patients received a liquid diet within 6 hours, were discharged in an average of 21 hours, and returned to full activity, usually within days. The only complication presented in this group of patients was an umbilical port seroma. Virtually all patients (97 percent) behaved as if only a colonoscopy had been performed. Pain at the trocar sites was managed with acetaminophen 600 mg by mouth as needed. CONCLUSION: Laparoscopic-monitored colonoscopic polypectomy allows patients to undergo removal of colonic polyps without a segmental resection. This less invasive procedure yields recovery times similar to that of colonoscopy alone, and the potential complications of a segmental resection are avoided. All polyps are examined by frozen section, and if a malignancy is encountered, a laparoscopic resection can be performed. PMID- 11005492 TI - Loss of standard type of CD44 expression in invaded area as a good indicator of lymph-node metastasis in colorectal carcinoma. AB - PURPOSE: Recent advances have made possible the treatment of small invasive colorectal cancer by means of polypectomy or endoscopic mucosal resection. CD44 expression in cancer cells was identified as an indicator of lymph-node metastasis, which could be evaluated in specimens removed by colonoscopy. METHODS: The correlation between lymph-node metastasis and the expression of standard-type CD44 in cancer cells was examined immunohistologically using the invaded cancer cells of 61 tissue samples of superficially invasive colorectal cancer. We defined the above as invasive cancer restricted within the colorectal wall. Of the 61 samples, 31 had submucosal invasion and 30 had muscular invasion. RESULTS: Standard-type CD44 expression in the area of invasion in cases with lymph-node metastasis was remarkably down-regulated. In 43 cases with no lymph node metastasis, 36 (83.7 percent) of patients had CD44 expression in invaded cells, whereas only two of 18 cases (11.1 percent) with lymph-node metastasis had expression of standard-type CD44 in the same area (P < 0.0001). A total of 69.6 percent (16/23) of patients with loss of standard-type CD44 expression in invaded sites were found to have positive metastasis in the lymph nodes. These results suggest that standard-type CD44 in invasive colon cancer cells could suppress metastasis to the regional lymph nodes. CONCLUSION: In cases of invasive colorectal cancer, the loss of standard-type CD44 expression in the invaded area is a sensitive marker for metastasis to the lymph nodes. Further investigation with larger patient groups is required to clarify the reliability of loss of standard-type CD44 expression as an indicator for additional surgery after endoscopic resection of submucosal invasive colorectal carcinoma. PMID- 11005493 TI - Results of neurophysiologic evaluation in fecal incontinence. AB - PURPOSE: Several methods of neurophysiologic assessment exist in the investigation of patients with fecal incontinence. However, the clinical significance of the information gained is uncertain. The aim of this prospective study was to evaluate the results of pudendal nerve terminal motor latency and fiber density in relation to clinical variables and manometric measurements. METHODS: Seventy-two patients with fecal incontinence (63 women; mean age, 62; range, 24-81 years) responded to a bowel questionnaire and underwent anorectal manovolumetry, anal ultrasonography, defecography, and electromyography, including pudendal nerve terminal motor latency and fiber density. RESULTS: Pudendal neuropathy (pudendal nerve terminal motor latency > 2.5 ms) was found in 46 percent and increased fiber density (> 1.7) in 82 percent. Pudendal neuropathy and increased fiber density were most common in patients with rectal prolapse or intra-anal intussusception. No difference was seen concerning anal resting and incremental pressures, rectal compliance, rectal sensibility or severity of incontinence in patients with unilateral, bilateral, or marked (> 4 ms) pudendal neuropathy vs. patients with normal pudendal nerve terminal motor latency. In contrast, patients with increased fiber density had lower incremental pressures (P < 0.05) and stated decreased rectal sensibility (P < 0.05) compared with those with normal fiber density. These differences were most pronounced in patients with neurogenic or idiopathic incontinence. CONCLUSIONS: Pudendal neuropathy and increased fiber density are common in patients with fecal incontinence. Fiber density but not pudendal nerve terminal motor latency was correlated with clinical and manometric variables. The severity of nerve injury correlated with anal motor and sensory function in patients with neurogenic or idiopathic incontinence. The routine use of pudendal nerve terminal motor latency in the assessment of patients with fecal incontinence can be questioned. PMID- 11005494 TI - Long-term results of electromyographic biofeedback training for fecal incontinence. AB - PURPOSE: The aim of this study was to examine the long-term results of electromyographic biofeedback training in fecal incontinence. METHODS: Thirty seven patients (1 male) received a customised program of 2 to 11 (median, 3) biofeedback training sessions with an anal plug electromyometer. Nine patients had persistent incontinence after anal sphincter repair, a further 8 patients had postsurgical or partial obstetric damage of the sphincter but no sphincter repair, 9 patients had neurogenic sphincter damage, and 11 patients were classified as having idiopathic fecal incontinence. Duration of voluntary sphincter contraction was measured by anal electromyography (endurance score) before and after treatment. A postal questionnaire was used to investigate the following variables: 1) subjective rating on a four-grade Likert-scale of the overall result of the biofeedback training; 2) incontinence score (maximum score is 18, and 0 indicates no incontinence); and 3) rating of bowel dissatisfaction using a visual analog scale (0 to 10). RESULTS: Twenty-two patients (60 percent) rated the result as very good (n = 8) or good (n = 14) immediately after the treatment period. Median endurance score improved from 1 to 2 minutes (P < 0.0001). Median incontinence score improved from 11 to 7, and bowel dissatisfaction rating improved from 5 to 2.8 (both P < 0.0001). After a median follow-up of 44 (range, 12-59) months, 15 patients (41 percent) still rated the overall result as very good (n = 3) or good (n = 12). The incontinence score did not change during follow-up. Median bowel dissatisfaction rating deteriorated from 2.8 to 4.2 but remained better than before treatment. Poor early subjective rating and the need for more than three biofeedback sessions were predictive of worsening during follow-up. CONCLUSION: We think it is encouraging that in this study biofeedback treatment for fecal incontinence with an intra-anal plug electrode resulted in a long-term success rate in nearly one-half of the patients. PMID- 11005495 TI - Treatment of impaired defecation associated with rectocele by behavorial retraining (biofeedback). AB - PURPOSE: Large rectoceles have been associated with symptoms of impaired rectal evacuation, often leading to rectocele repair. However, these symptoms, or the anatomic abnormality, may be caused, at least in part, by a primary disturbance of rectoanal coordination. This study aimed to determine the efficacy of biofeedback therapy in such patients. METHODS: Thirty-two female patients (median age, 52 years) complaining of impaired rectal evacuation and with a rectocele greater than 2 cm at proctography were evaluated by structured questionnaire before, immediately after treatment, and at follow-up. Physiologic and proctographic findings were related to outcome. RESULTS: Immediate results were available in 32 patients and medium-term follow-up (median, 10; range, 2-30 months) in 25 patients. At follow-up 14 (56 percent) patients felt a little and 4 (16 percent) patients felt major improvement in symptoms, including 3 (12 percent) with complete symptom relief. Immediately after biofeedback there was a modest reduction in need to strain (from 72 to 50 percent), feeling of incomplete evacuation (from 78 to 59 percent), need to assist defecation digitally (from 84 to 63 percent), and need to use an evacuant (from 47 to 28 percent), and this was maintained at follow-up. Bowel frequency was significantly normalized at follow up (P = 0.02). Pretreatment presence of symptoms of digitally assisting defecation, pelvic floor incoordination, and proctographic rectocele size and contrast trapping, did not predict outcome. CONCLUSIONS: Behavioral therapy, including biofeedback, leads to major symptom relief in a minority, and partial symptom relief in a majority, of patients with a feeling of impaired defecation and the presence of a large rectocele. Residual symptoms are common. Biofeedback may be a reasonable first-line treatment for such patients. PMID- 11005496 TI - Recurrent rectal prolapse: what is the next good option? AB - PURPOSE: The aim of this study was to assess the clinical and functional outcome of surgery for recurrent rectal prolapse and compare it with the outcome of patients who underwent primary operation for rectal prolapse. METHODS: All patients who underwent surgery for rectal prolapse were evaluated for age, gender, procedure, anorectal manometry and electromyography findings, and morbidity. The results for patients who underwent surgery for recurrent rectal prolapse were compared with a group of patients matched for age, gender, surgeon, and procedure who underwent primary operations for rectal prolapse. RESULTS: A total of 115 patients underwent surgery for rectal prolapse. Twenty-seven patients, 10 initially operated on at this institution and 17 operated on elsewhere, underwent surgery for recurrent rectal prolapse. These 27 patients were compared with 27 patients with primary rectal prolapse operated on in our department. In the recurrent rectal prolapse group, prior surgery included rectopexy in 7 patients, Delorme's procedure in 7 patients, perineal rectosigmoidectomy in 7 patients, anal encirclement procedure in 4 patients, and resection rectopexy in 2 patients. Operations performed for recurrence were perineal rectosigmoidectomy in 14 patients, resection rectopexy in 8 patients, rectopexy in 2 patients, pelvic floor repair in 2 patients, and Delorme's procedure in 1 patient. There were no statistically significant differences between the groups in preoperative incontinence score (recurrent rectal prolapse, 13.6 +/- 7.8 vs. rectal prolapse, 12.7 +/- 7.2; range, 0-20) or manometric or electromyography findings, and there were no significant differences in mortality (0 vs. 3.7 percent), mean hospital stay (5.4 +/- 2.5 vs. 6.9 +/- 2.8 days), anastomotic complications (anastomotic stricture (0 vs. 7.4 percent), anastomotic leak (3.7 vs. 3.7 percent) and wound infection (3.7 vs. 0 percent), postoperative incontinence score (2.8 +/- 4.8 vs. 1.5 +/- 2.7), or recurrence rate (14.8 vs. 11.1 percent) between the two groups at a mean follow-up of 23.9 (range, 6-68) and 22 (range, 5-55) months, respectively. The overall success rate for recurrent rectal prolapse was 85.2 percent. CONCLUSION: The outcome of surgery for rectal prolapse is similar in cases of primary or recurrent prolapse. The same surgical options are valid in both scenarios. PMID- 11005497 TI - Primary colorectal lymphoma. AB - PURPOSE: The purpose of this study was to review the clinical presentation and characteristics of primary colorectal lymphoma, analyze the prognostic factors, and assess the results of treatment with adjuvant chemotherapy. METHODS: We identified 37 cases at our institution between 1980 and 1996. They comprised 0.48 percent of all cases of colon malignancies (37/7,658) during this period. The following clinical information was obtained: age, gender, signs and symptoms, tumor site, tumor size, histology grade, pathology, and adjuvant chemotherapy. RESULTS: The most common presenting signs and symptoms were abdominal pain (62 percent), abdominal mass (54 percent), and weight loss (43 percent). The most frequent site of involvement was the cecum (45 percent). Histologically, 29 (78 percent) were classified as high-grade, and 8 (22 percent) as intermediate-grade to-low-grade lymphoma. Nine (24.3 percent) of the cases were Stage EI, 23 (62.2 percent) were Stage EII, and 5 (13.5 percent) were Stage EIV. Twenty-one (57 percent) cases received adjuvant chemotherapy. The five-year survival rate was 33 percent for all patients and 39 percent for patients treated with combination chemotherapy. Overall median survival time was 24 months and 36 months for those with adjuvant chemotherapy. Only histology grade, among the factors examined, was a significant prognostic factor for survival. The mean survival time of the patients with Stage II disease who received chemotherapy was 117.4 months, and it was 47.9 months for the patients with Stage II disease who did not received chemotherapy. CONCLUSIONS: In our retrospective study high-grade lymphoma was the only significant adverse prognostic factor for survival. Receiving adjuvant chemotherapy significantly improved survival in patients with Stage II disease. Patients with diffuse large-cell type had better survival than patients with small noncleaved-cell type in Stage II high-grade lymphoma. PMID- 11005498 TI - Effects of rectal mobilization and lateral ligaments division on colonic and anorectal function. AB - PURPOSE: Colonic and anorectal function are altered after posterior rectopexy. The aim of this randomized, prospective study was to evaluate the effects of rectal mobilization and division of the lateral ligaments on colonic and anorectal function. METHODS: Posterior rectopexy was performed in 18 patients with complete rectal prolapse. Anal manometry and measurement of rectal compliance, total and segmental colonic transit time, constipation score, and defecation frequency were performed preoperatively and three months postoperatively. Ligaments were divided in ten patients. RESULTS: Mean preoperative total transit time was similar between the two patient groups and doubled postoperatively (P = 0.03). Mean postoperative segmental transit time increased by a factor of 1.7 in segments I (ascending colon) and II (descending colon) and by a factor of 2.3 in segment III (rectosigmoid). The same pattern was found in both groups. Mean resting pressure decreased after division of the lateral ligaments and increased after preservation. Mean rectal compliance decreased after division of the ligaments and increased when they were preserved. Mean postoperative constipation score differed little from the preoperative score. Mean defecation frequency was decreased in the group with the ligaments preserved and increased in the group with the ligaments divided. None of the effects of rectal mobilization or division of the lateral ligaments on anorectal function reached statistical significance. CONCLUSION: Rectal mobilization had a statistically significant effect on colonic function. Total and segmental colonic transit times doubled. The effects on anorectal function were not significant. Division of the lateral ligaments did not significantly influence postoperative functional outcome. PMID- 11005499 TI - Transanally injected triamcinolone acetonide in levator syndrome. AB - PURPOSE: Several treatments are used for the treatment of levator syndrome, such as rectal massage, biofeedback, and galvanic stimulation. But their effects are inconsistent, and multiple treatment sessions are usually required. Triamcinolone acetonide and lidocaine mixture was injected locally into the tenderest point in levator syndrome under the hypothesis that levator syndrome is caused by tendinitis of pelvic floor musculature. METHODS: A mixture of 40 mg of triamcinolone acetonide and 1 ml of 2 percent lidocaine was injected into the tenderest point transanally in 104 patients (33 males; mean age, 51 years) with levator syndrome from December 1996 to May 1998 at Daehang Clinic. Additional injections were repeated at two-week intervals to a maximum of three injections in cases of poor response. Follow-up was performed prospectively concerning patient's perception of pain level using a visual analog scale. Depending on the response, the patients were classified into four groups: pain-free, good, fair, and no response. More than 50 percent pain reduction was classified as "good," and less than 49 percent reduction as "fair." RESULTS: The injection regions, where the tenderest points were identified on digital rectal compression, were left anterior anal canal in 71.2 percent of patients, right anterior in 3.8 percent of patients, and posterior in 25 percent of patients. The results of treatment were as follows: at three months after injection, response was classified as pain-free in 36.8 percent of patients, good in 35 percent of patients, fair in 19.5 percent of patients, and no response in 8.7 percent of patients; at six months the response was pain-free in 30.1 percent of patients, good in 46.5 percent of patients, fair in 18.2 percent of patients, and no response in 5.2 percent of patients. Most patients, except 8.7 percent at three months and 5.2 percent at six months, experienced treatment benefits. There were no complications during the follow-up periods. CONCLUSION: Transanal injection of triamcinolone acetonide and lidocaine mixture into the tenderest point is such a simple, safe, and very effective modality that it can be recommended as a primary therapy for levator syndrome. PMID- 11005500 TI - Bile carcinoembryonic antigen levels and occult hepatic metastases from colorectal cancer. AB - PURPOSE: Up to 30 percent of patients will have occult hepatic metastases at the time of curative surgery for colorectal cancer. The ability to predict this group of patients would allow better targeting of appropriate therapy. It has been shown previously that patients with overt hepatic metastases have significantly high levels of carcinoembryonic antigen in gallbladder bile compared with serum levels. The aim of this study was to assess the accuracy of bile carcinoembryonic antigen levels taken at the time of operation in predicting patients with occult hepatic metastases. METHODS: Bile and serum carcinoembryonic antigen samples were collected from 37 patients undergoing surgery for colorectal cancer, 26 of whose procedures were deemed curative and who were followed up for a median of 63.5 months. RESULTS: Twelve patients were alive with no evidence of recurrent disease, and two had recurrent disease, whereas 12 died of disease. The median (interquartile range) serum carcinoembryonic antigen in the disease-free group was 2.8 (1.1-6.1) ng/ml, and in the recurrent group it was 6.35 (4.3-30) ng/ml (P = 0.006), whereas bile carcinoembryonic antigen in the disease-free group was 7 (5-39) ng/ml as compared with 31 (5-383.7) ng/ml in the recurrent group (P = 0.210). The accuracy of serum carcinoembryonic antigen in predicting occult hepatic metastases was 77 percent compared with 72 percent for bile carcinoembryonic antigen. CONCLUSION: Intraoperative bile carcinoembryonic antigen levels are no more accurate than serum carcinoembryonic antigen levels in predicting occult hepatic metastases in patients undergoing potentially curative colorectal cancer surgery. PMID- 11005501 TI - Impact of previous surgery on time taken for incision and division of adhesions during laparotomy. AB - PURPOSE: Adhesions from previous surgery constitute a major problem in general surgery. Much is known of the consequences of adhesions in terms of morbidity, but there has been little documentation of the effect of previous surgery on the time taken to reoperate. METHODS: Information on incision time and division of adhesion time was recorded on 120 patients (89 elective cases and 31 emergencies) undergoing midline laparotomy under the care of a single colorectal surgeon. RESULTS: Fifty-one percent of elective and 71 percent of emergency cases had previously had abdominal surgery. Previous surgery prolonged the median incision time from 5 (range, 3-10) to 8 (range, 4-39) minutes (P < 0.001) and the median division of adhesion time from 0 (range, 0-30) to 15 (range, 0-12) minutes (P < 0.0001). CONCLUSIONS: Previous surgery significantly increases the operating time during subsequent surgery by a median of 18 minutes. This information may be of use in the planning of surgical workload and highlights the needs for cost effective adhesion prevention strategies. PMID- 11005502 TI - Fournier's gangrene: changing face of the disease. AB - PURPOSE: Our experience with ten cases of Fournier's gangrene prompted us to review the related literature to highlight the current status of the disease. METHODS: Data from ten patients with the diagnosis of Fournier's gangrene treated at our center from January 1997 until December 1998 were analyzed. These patients were treated by aggressive resuscitation, triple antibiotics, and urgent surgery. The English-language medical literature for the past 30 years was reviewed. RESULTS: The epidemiologic features of our patients were similar to those reported in other recent studies. Mortality rate was 20 percent. Currently, the disease affects both genders and a wide range of ages, has a more insidious onset than in the past, and is not idiopathic. Associated systemic disorders (diabetes, alcoholism, and immunosuppression) are common. Perianal infection is the commonest cause and is associated with more moribund features. CONCLUSION: The epidemiology of Fournier's gangrene is changing from its original description. Population aging worldwide--as a result of improving health care--and therefore the increasing prevalence of associated medical disorders may explain these changes. These factors may also explain the consistently high mortality rate during more recent years, masking any survival benefits from improved medical care. Better understanding of the pathophysiology has reduced the ratio of idiopathic cases to a minimum. PMID- 11005503 TI - Hyperplastic polyps: "more than meets the eye"? Report of sixteen cases. AB - The vast majority of hyperplastic polyps are small, left-sided, and inconsequential in nature. However, hyperplastic polyps that are large, right sided, mixed, and found in association with a family history of carcinoma may represent an "atypical" group, and their clinical significance is uncertain. We believe that these atypical lesions should not be lumped together with the common variety of diminutive hyperplastic polyps. Rather, when such hyperplastic polyps are encountered, they should be excised and the patient should be placed on regular colonoscopic surveillance. PMID- 11005504 TI - Synchronous colorectal and renal carcinomas: a noteworthy clinical entity. Report of five cases. AB - PURPOSE: This study was designed to highlight the incidence of synchronous colorectal and renal carcinomas and to review the literature on that issue. The case reports of five patients who presented with synchronous colorectal and renal cell carcinomas are presented. METHODS: A retrospective study, using systematic medical chart review, analyzed the cases of all patients who underwent large bowel resection for colorectal cancer in our department between December 1996 and December 1998. RESULTS: Among 103 patients who underwent colorectal surgery during that period, five cases of synchronous colorectal and renal carcinomas were detected (4.85 percent). CONCLUSIONS: Based on our findings, we recommend the routine use of preoperative imaging studies to exclude synchronous asymptomatic renal lesions in patients presenting with colorectal cancer. PMID- 11005505 TI - Gastrointestinal stromal tumor of the rectum with activating mutation of c-kit: report of a case. AB - Nonepithelial malignancies of the large bowel are rare. A new disease entity, gastrointestinal stromal tumors, has attracted attention among primary mesenchymal tumors of the gastrointestinal tract. Here we present a case of spindle-cell sarcoma of the rectum, lacking either smooth muscle cells or neural elements. Immunohistochemical findings and sequencing of the c-kit proto-oncogene diagnosed this tumor as a malignant gastrointestinal stromal tumor of the rectum. PMID- 11005506 TI - Thermal imaging in the detection of bowel ischemia. AB - PURPOSE: The aim of this study was to introduce thermal imaging in the intraoperative detection of bowel ischemia by comparing thermal imaging with conventional techniques in detecting acutely ischemic bowel, using histologic evidence for intestinal necrosis as the standard. METHODS: A prospective study was performed using a porcine model. Laparotomy was performed on four pigs under general anesthesia. A 25-cm segment of mid jejunum was tagged with proximal and distal sutures, and its mesentery was ligated and divided. Thermal imaging, visual inspection, Doppler ultrasound, and fluorescence with Wood's lamp after fluorescein were used to estimate the extent of bowel ischemia five minutes after ligation of the mesentery. Measurements were taken in reference to both the proximal and distal tags to obtain two data points per animal for each method. After two hours of warm ischemia, the jejunum was harvested and sectioned longitudinally. Comparisons were made between the estimated region of necrosis for each method and microscopic evidence of necrosis. RESULTS: Visual inspection was the only method unable to detect a difference between vascularized and devascularized bowel for each of the eight data points. Fluorescein dye missed 3 cm of ischemic bowel. Doppler ultrasound and thermal imaging were 100 percent sensitive for necrotic bowel, with thermal imaging overestimating necrosis to a greater extent than Doppler ultrasound. The positive predictive value of fluorescein dye, Doppler ultrasound, and thermal imaging for determining nonviable bowel was 91.8, 80.8, and 69.5 percent, respectively. CONCLUSIONS: Thermal imaging has the potential to be a useful adjunct in the intraoperative determination of bowel ischemia. Further studies are indicated to study this technique. PMID- 11005507 TI - Laparostomy for severe intra-abdominal infection complicating colorectal disease. PMID- 11005508 TI - Escape mechanisms in tumor immunity: a year 2000 update. AB - The current consensus of opinion has it that most or possibly all tumors, spontaneous as well as induced, are immunogenic, expressing antigens in a form recognizable by the host immune system. Accordingly, in order to progress, tumors have to evolve strategies for evading immune responses. The purpose of this review is to consider the current status of knowledge concerning these different tumor escape strategies. It represents an update of an article originally published in this journal in 1997 (Pawelec, Zeuthen, and Kiessling, 1997). Therefore, it focuses mostly on publications that have appeared since then, illustrating the impressive accumulation of new data since that time and the importance currently attributed to studies of tumor escape from the immune response. PMID- 11005509 TI - Vascular endothelial growth factor and osteopontin in tumor biology. AB - Tumor growth and metastasis are angiogenesis-dependent and tumor angiogenesis is a result of complex interplay of positive and negative regulators. Vascular endothelial growth factor (VEGF) occupies a particular place among the positive regulators of angiogenesis due to its potency and specificity for endothelial cells. VEGF upregulates several molecules such as growth factors, adhesion molecules, proteases, and protease receptors and it actually induces microvascular hyperpermeability, resulting in activation of thrombin from prothrombin. Osteopontin (OPN) is a secreted arginine-glycine-asparic acid (RGD) containing phosphoprotein and it contains a predicted thrombin cleavage site. OPN binds to several integrins and CD44 variants. OPN has diverse functions such as cell adhesion, chemoattraction, and immunomodulation, and it induces endothelial cell migration and upregulates endothelial cell migration induced by VEGF. OPN expression is upregulated in human carcinomas. This review documents the functional roles of VEGF and OPN in angiogenesis and their clinical significance in tumor biology. PMID- 11005510 TI - Differentiation and metastasis in melanoma. AB - The incidence of melanoma has been rising steadily during the last four decades and is now among the highest of all human cancers. As for most tumors, malignancy and metastatic spreading represent the deadly aspects of a tumor that, if eradicated before becoming invasive, can be easily cured. In fact, melanoma metastatic to regional lymph nodes carries a poor prognosis, and distant metastatic melanoma becomes incurable. Because traditional forms of chemotherapy have little effect on this type of tumor, differentiation therapy has been considered as a possible alternative, based on the consideration that malignancy and differentiation are usually inversely related. However, the relationship between these two factors turned out to be more complex for melanoma cells, and in some murine model systems it has been found that differentiated cells, although less tumorigenic, could be even more metastatic. No clear correlation has been reported between (epi)genetic changes induced by differentiating drugs and the increased malignant phenotype. This review examines what is known to date about differentiation state and metastatic ability of melanomas, and also reports some novel data with the B16 mouse melanoma model system. PMID- 11005511 TI - Impact of viral oncogenesis on responses to anti-cancer drugs and irradiation. AB - The view that chemical or physical oncogenesis and tumor therapy resistance represent different parts of common cellular alterations gained considerable attractiveness, because it explains the inherent unreponsiveness of many tumors. Viruses are potent oncogenes and are causally linked to approximately one-fifth of all human malignancies. Whether viral oncogenesis exerts comparable effects was less clear. Recent progress in experimental research provided ample evidence that viruses affect response of tumor cells toward anti-cancer drugs and irradiation. Resistance to cytostatic drugs and radiation develops by alterations at the drug-target sites (i.e., DNA or specific target proteins), upstream (i.e., detoxification mechanisms), or downstream of them (i.e., programmed cell death). Viruses interfere with specific cellular genes at these three levels. Viral proteins induce the expression and expression of drug resistance genes, that is, MDR1, DHFR, or CAD. Viral interactions with the tumor suppressor genes (p53, pRB) abrogate cell cycle arrests and disturb DNA repair of drug- and radiation-induced DNA lesions. The readiness to commit cellular suicide (apoptosis) is also affected by viral genes. The connection between viral oncogenesis and the response of tumor cells to treatment adds a new dimension to tumor biology and may have important consequences for oncological treatment modalities in the future. PMID- 11005512 TI - Extended slide trochanteric osteotomy for revision total hip arthroplasty. AB - BACKGROUND: The purpose of this study was to assess the rate of union, time to union, and complications associated with the extended slide trochanteric osteotomy. We also evaluated how outcomes were influenced by the preoperative cortical-bone thickness, the preoperative cancellous-bone quality of the greater trochanter, the number of cables used to reattach the trochanteric osteotomy fragment, and the use of cortical strut augmentation. METHODS: We reviewed the results for forty-six hips in forty-five patients who underwent a revision total hip arthroplasty with an extended slide trochanteric osteotomy between December 1991 and December 1996. Twenty-three patients were men, and twenty-two were women; the mean age at the time of the operation was 66.3 years. Two hips had an isolated acetabular revision, fifteen had an isolated femoral revision, and twenty-nine had acetabular and femoral revisions. One patient (one hip) was lost to follow-up. RESULTS: At a mean of forty-four months after the operation, the rate of union of the distal osteotomy site was 98 percent (forty-four of forty five hips), with no change in the femoral component position. The time to union was not significantly correlated with the number of cables, the preoperative cortical-bone thickness, or the preoperative cancellous-bone quality of the greater trochanter. Interestingly, the time to bridging-callus union was significantly longer in the hips with a strut allograft than in the hips without a strut allograft (p = 0.04, t test for independent samples). Two fractures of the osteotomy fragment occurred, but neither necessitated another revision. CONCLUSIONS: The extended slide trochanteric osteotomy allows extensive acetabular and femoral exposure, facilitates removal of distal cement or a well fixed porous-coated stem, and allows reliable reattachment and healing of the trochanteric fragment. PMID- 11005513 TI - Capsulolabral augmentation for the the management of posteroinferior instability of the shoulder. AB - BACKGROUND: Posteroinferior instability of the shoulder has been associated with capsular laxity. The purposes of the present study were to describe the pathological morphology of the posteroinferior aspect of the glenolabral fossa in patients with primary posteroinferior instability and to prospectively examine the efficacy of managing this instability with use of an arthroscopic posteroinferior capsulolabral augmentation procedure. METHODS: Forty-one patients who had posteroinferior instability of the shoulder were managed with an arthroscopic shift of the posteroinferior aspect of the capsule to the adjacent labrum and were followed for a minimum of twelve months. Thirty-two patients had a primary procedure, and nine had a revision procedure. The mean duration of follow-up was twenty-eight months (range, twelve to sixty-nine months). All of the patients had presented with a symptomatic, positive finding on the jerk test and had participated in a minimum of six months of rehabilitation that had failed to relieve the symptoms. The patients were evaluated prospectively with a motion and stability examination and the Simple Shoulder Test. In addition, they completed the Short Form-36 Health Survey (SF-36) and a questionnaire on the outcome of treatment. RESULTS: Lesions affecting the posteroinferior aspect of the glenolabral concavity were seen in thirty-four patients (83 percent): five had labral detachment, seven had chondral or labral erosion, nine had capsular and synovial stripping, and thirteen had a labral split or tear. The mean score (and standard deviation) on the Simple Shoulder Test improved from 5.5 +/- 3.4 points to 8.1 +/- 3.3 points (p = 0.0023), and two of the eight SF-36 parameters improved significantly (p < 0.05). Conversely, nineteen patients who were receiving Workers' Compensation did not show any improvement in either of the two parameters. Thirty-five patients had improved stability of the shoulder, and the findings on all physical examinations had improved significantly (p < 0.0001). Twenty-eight patients had a perception of residual stiffness; this finding was in contrast to the mean score on the flexibility examination, which had not changed significantly at the time of the latest follow-up. CONCLUSIONS: Posteroinferior instability of the shoulder is associated not only with capsular laxity but also with well defined lesions of the glenolabral concavity. Arthroscopic capsulolabral augmentation to reduce posterior capsular laxity and to restore the depth of the glenolabral concavity has been shown to be effective treatment of this condition after a mean duration of follow-up of twenty-eight months. PMID- 11005514 TI - Arthroplasty of the hip in patients with aplastic anemia. AB - BACKGROUND: Patients with aplastic anemia are now living longer and therefore are at increased risk for the development of osteonecrosis of the hip. However, studies on the results of arthroplasty for the treatment of osteoarthritis of the hip in patients with aplastic anemia are lacking. METHODS: Twenty-six primary hip prostheses (one bipolar prosthesis fixed with cement, two bipolar prostheses fixed without cement, three hybrid total hip prostheses, and twenty total hip prostheses fixed without cement) were implanted, between March 1990 and May 1992, in nineteen patients who had been diagnosed with aplastic anemia. A specific prospective protocol was followed for the perioperative transfusion of platelets and blood. Twenty-five hips were replaced because of osteonecrosis of the femoral head, and one was replaced because of a femoral neck fracture. The patients were followed prospectively, with preoperative and serial postoperative Harris hip ratings as well as radiographs, for a minimum of six years or until death. RESULTS: No patient had excessive perioperative bleeding or a postoperative infection. After a mean duration of follow-up of seventy-nine months (range, seventy-two to ninety-five months), two patients had died with the original implant in place. No patients were lost to follow-up. The mean Harris hip score was 55 points (range, 42 to 68 points) preoperatively and 87 points (range, 56 to 95 points) at the time of the latest follow-up. At the time of this writing, no hip had been revised. One patient with a bipolar prosthesis had radiographic evidence of femoral loosening and will probably require revision. A second patient had some medial protrusion of a bipolar prosthesis, with mild symptoms. All of the acetabular components that had been fixed without cement and all of the other femoral components appeared to be stable on radiographs after a minimum of seventy-two months of follow-up. CONCLUSIONS: Total hip arthroplasty can be performed safely in patients with aplastic anemia. In the present intermediate term study, the durability of implant fixation was maintained and the clinical results demonstrated a sustained increase in function of the hip. PMID- 11005515 TI - Noninvasive imaging predicts failure load of the spine with simulated osteolytic defects. AB - BACKGROUND: The clinical management of lytic tumors of the spine is currently based on geometric measurements of the defect. However, the mechanical behavior of a structure depends on both its material and its geometric properties. Quantitative computed tomography and dual-energy x-ray absorptiometry were investigated as noninvasive tools for measuring the material and geometric properties of vertebrae with a simulated lytic defect. From these measures, yield loads were predicted with use of composite beam theory. METHODS: Thirty-four fresh-frozen cadaveric spines were segmented into functional spinal units of three vertebral bodies with two intervertebral discs at the thoracic and lumbar levels. Lytic defects of equal size were created in one of three locations: the anterior, lateral, or posterior region of the vertebra. Each spinal unit was scanned with use of computed tomography and dual-energy x-ray absorptiometry, and axial and bending rigidities were calculated from the image data. Each specimen was brought to failure under combined compression and forward flexion, and the axial load and bending moment at yield were recorded. RESULTS: Although the relative defect size was nearly constant, measured yield loads had a large dispersion, suggesting that defect size alone was a poor predictor of failure. However, image-derived measures of structural rigidity correlated moderately well with measured yield loads. Furthermore, with use of composite beam theory with quantitative computed tomography-derived rigidities, vertebral yield loads were predicted on a one-to-one basis (concordance, r(c) = 0.74). CONCLUSIONS: Although current clinical guidelines for predicting fracture risk are based on geometric measurements of the defect, we have shown that the relative size of the defect alone does not account for the variation in vertebral yield loads. However, composite beam theory analysis with quantitative computed tomography-derived measures of rigidity can be used to prospectively predict the yield loads of vertebrae with lytic defects. CLINICAL RELEVANCE: Image-predicted vertebral yield loads and analytical models that approximate loads applied to the spine during activities of daily living can be used to calculate a factor of fracture risk that can be employed by physicians to plan appropriate treatment or intervention. PMID- 11005516 TI - Total knee arthroplasty after high tibial osteotomy. A comparison study in patients who had bilateral total knee replacement. AB - BACKGROUND: The outcome of total knee replacement after high tibial osteotomy remains uncertain. We hypothesized that the results of total knee replacement with or without a previous high tibial osteotomy are similar. METHODS: The results of a consecutive series of thirty-nine bilateral total knee arthroplasties performed with cement at an average of 8.7 years after unilateral high tibial osteotomy were reviewed. There were twenty-seven men and twelve women. Preoperatively, the knee scores according to the system of the Knee Society were similar for all of the knees; however, valgus alignment and patella infera were more common in the knees with a previous high tibial osteotomy. Bilateral total knee replacement was staged in seven patients and was simultaneous in thirty-two patients. The results of the total knee arthroplasties were retrospectively reviewed with respect to the knee and function scores according to the system of the Knee Society, the radiographic findings, and the complications. RESULTS: Intraoperatively, no notable differences were identified in the number of medial, lateral, or lateral patellar releases required. However, less lateral tibial bone was resected in the group with a previous high tibial osteotomy (average, 3.3 millimeters) than in the group without a high tibial osteotomy (average, 7.5 millimeters). The average duration of follow-up was 7.5 years (range, three to sixteen years) in the group with a previous high tibial osteotomy and 6.8 years (range, two to ten years) in the group without a high tibial osteotomy. At the time of the final follow-up, the knee and function scores were similar for the two groups (89.0 and 81.0 points, respectively, for the group with a previous high tibial osteotomy, and 89.6 and 83.9 points, respectively, for the group without a high tibial osteotomy). Although more knees were free of pain in the group without a previous high tibial osteotomy (thirty six) than in the group with a previous osteotomy (thirty-three), this difference was not found to be significant with the numbers available (p = 0.4810). Knee alignment and stability, femoral and tibial component alignment, and range of motion also were similar in both groups postoperatively. One allpolyethylene tibial component was revised in the high tibial osteotomy group. Two knees in each group required manipulation. There were no deep infections. CONCLUSIONS: While patients with a previous high tibial osteotomy may have important differences preoperatively, including valgus alignment, patella infera, and decreased bone stock in the proximal part of the tibia, the present study suggests that the clinical and radiographic results of primary total knee arthroplasty in knees with and without a previous high tibial osteotomy are not substantially different. In our relatively small group of patients, the previous high tibial osteotomy had no adverse effect on the outcome of the subsequent total knee replacement. PMID- 11005517 TI - Semiconstrained total elbow arthroplasty for ankylosed and stiff elbows. AB - BACKGROUND: Total elbow arthroplasty can be a valuable option for the treatment of ankylosed or very stiff elbows. METHODS: A semiconstrained total elbow arthroplasty was performed in thirteen patients (fourteen elbows) with a preoperative range of elbow motion of 30 degrees or less. Nine elbows were fused or ankylosed preoperatively. The mean age at the time of the surgery was fifty years (range, twenty-four to seventy-nine years). The etiology of the stiffness was trauma for eleven elbows, juvenile rheumatoid arthritis for two, and rheumatoid arthritis for one. RESULTS: After a mean duration of follow-up of sixty-three months, the result was excellent for four elbows, good for four, fair for one, and poor for five, according to the Mayo elbow performance score. The mean arc of flexion improved from 7 degrees (range, 0 to 30 degrees) preoperatively to 67 degrees (range, 10 to 115 degrees) after the surgery. The most important factor that influenced the final result was the presence of ectopic bone surrounding the elbow joint. There were seven complications. Infection developed in five elbows. Three elbows had a superficial infection, which did not compromise the final result in two and which was treated with a myocutaneous flap in one with skin necrosis, with an excellent result. Deep infection developed in two other elbows. Both had an unsatisfactory result, one after implant removal and one after several debridements and retention of the prosthesis. Two patients sustained a fracture because of a loose component, and the prosthesis was revised. Four patients who lost motion within the first month following the surgery had a manipulation under anesthesia. CONCLUSIONS: Semiconstrained total elbow arthroplasty is a useful option for patients with an ankylosed or a very stiff elbow and results in a considerable improvement of motion. Because of the nature of the underlying pathology, complications, including reoperation, are frequent, but the risk can be lessened by careful preoperative planning and surgical technique. Replacement is the preferred option in patients who are more than sixty years of age, but it is also a good choice in younger patients if there is no other viable option. PMID- 11005518 TI - Osteochondromas of the distal aspect of the tibia or fibula. Natural history and treatment. AB - BACKGROUND: There is little information on the natural history or treatment of osteochondromas arising from the distal aspect of either the tibia or the fibula. It is believed that there is a risk of deformation of the ankle if these exostoses are left untreated or if the physis or neurovascular structures are injured during operative intervention. METHODS: We reviewed the records of twenty three patients who had been treated for osteochondroma of the distal aspect of the tibia or fibula between 1980 and 1996. Four of the patients had hereditary multiple cartilaginous exostoses. There were seventeen male and six female patients, and the average age at the time of presentation was sixteen years (range, eight to forty-eight years). RESULTS: Preoperative radiographs showed evidence of plastic deformation of the fibula in eleven patients who had a large osteochondroma. Four patients elected not to have an operation. The tumor was excised in nineteen patients. Postoperatively, all nineteen patients had a Musculoskeletal Tumor Society score of 100 percent for function of the lower extremity with pain-free symmetrical and unrestricted motion of the ankle at the latest follow-up examination. Partial remodeling of the tibia and fibula gradually diminished the asymmetry of the ankles in all nineteen operatively managed patients; however, the remodeling was most complete in the younger patients. Pronation deformities of the ankle did not change after excision of the tumor. Complications of operative treatment included four recurrences (only three of which were symptomatic), one sural neuroma, one superficial wound infection, and one instance of growth arrest of the distal aspects of the tibia and fibula. CONCLUSIONS: Osteochondromas of the distal and lateral aspects of the tibia were more often symptomatic than those of the distal aspect of the fibula; they most commonly occurred in the second decade of life with ankle pain, a palpable mass, and unrestricted ankle motion. Untreated or partially excised lesions in skeletally immature patients may become larger and cause plastic deformation of the tibia and fibula and a pronation deformity of the ankle. Ideally, operative intervention should be delayed until skeletal maturity, but, in symptomatic patients, partial excision preserving the physis may be necessary for the relief of symptoms and the prevention of progressive ankle deformity. However, partial excision is associated with a high rate of recurrence, so a close follow-up is required. Skeletally mature patients who are symptomatic may require excision of the tumor. PMID- 11005519 TI - Atraumatic osteonecrosis of the knee. AB - BACKGROUND: The purposes of this study were to define the clinical, demographic, and radiographic patterns of atraumatic osteonecrosis of the distal part of the femur and the proximal part of the tibia at presentation and to report the outcome of treatment of this condition. METHODS: Two hundred and forty-eight knees in 136 patients who were younger than the age of fifty-five years were treated at our institution between July 1, 1974, and September 15, 1998, for atraumatic osteonecrosis of the distal part of the femur or the proximal part of the tibia, or both. Demographic and radiographic features were characterized. The results of nonoperative treatment, core decompression, arthroscopic debridement, and total knee arthroplasty were evaluated. RESULTS: There were 106 female patients and thirty male patients, and their mean age was thirty-six years (range, fifteen to fifty-four years) at the time of diagnosis. One hundred and one patients (74 percent) had involvement of other large joints, with eighteen (13 percent) presenting initially with knee symptoms. One hundred and one patients (74 percent) had a disease that affected the immune system; sixty-seven of them had systemic lupus erythematosus. One hundred and twenty-three patients (90 percent) had a history of corticosteroid use. Technetium-99m bone-scanning missed lesions in sixteen (29 percent) of fifty-six knees. Eight (20 percent) of forty-one initially symptomatic knees treated nonoperatively had a successful clinical outcome (a Knee Society score of at least 80 points and no additional surgery) at a mean of eight years. The knees that remained severely symptomatic for three months were treated with either core decompression (ninety-one knees) or total knee arthroplasty (seven knees). Seventy-two (79 percent) of the ninety one knees treated with core decompression had a good or excellent clinical outcome at a mean of seven years. Efforts to avoid total knee arthroplasty with repeat core decompression or arthroscopic debridement led to a successful outcome in fifteen (60 percent) of twenty-five knees. Thirty-four (71 percent) of forty eight knees treated with total knee arthroplasty had a successful clinical outcome at a mean of nine years. CONCLUSIONS: Atraumatic osteonecrosis of the knee predominantly affects women, and in our study it was associated with corticosteroid use in 90 percent of the patients. Evaluation should include standard radiographic and magnetic resonance imaging of all symptomatic joints. Prognosis was negatively related to large juxta-articular lesions. Nonoperative treatment should be reserved for asymptomatic knees only. Core decompression was successful (a Knee Society score of at least 80 points and no additional surgery) in 79 percent of the knees in which the disease was in an early stage. Total knee arthroplasty was successful in only 71 percent of the knees. PMID- 11005520 TI - Hybrid total hip arthroplasty with a precoated offset stem. Four to nine-year results. AB - BACKGROUND: Use of modern cementing techniques for fixation of femoral components in total hip arthroplasty has had excellent clinical and radiographic results in most patients. However, several authors have described early loosening of femoral components with roughened and precoated finishes. The purpose of this study was to examine the performance of the precoated Iowa stem, which has increased offset, and to compare the results with those of another cemented precoated femoral component with standard offset used at our institution. METHODS: We carried out a prospective analysis of 102 primary hybrid total hip arthroplasties (a cementless acetabular component and a cemented femoral component) performed with use of the Iowa femoral component in ninety-five patients at our institution. The Iowa stem was used in hips that required greater offset than is available with standard stems as determined by preoperative templating. The average age of the patients at the time of the index procedure was sixty-nine years. Sixteen patients (seventeen hips) died before the forty-eight-month minimum follow-up period had elapsed. Two patients were lost to follow-up, and radiographic follow-up was incomplete for one. The mean duration of clinical and radiographic follow-up of the remaining eighty-two hips in the seventy-six surviving patients was sixty-five months (range, forty-eight to 104 months). RESULTS: The average preoperative Harris hip score of 47 points (range, 16 to 69 points) improved to an average of 87 points (range, 24 to 100 points) at the time of the review. Two hips underwent femoral component revision. Four femoral stems were radiographically loose at an average of thirty-four months. Femoral osteolysis was seen in five hips (6 percent) at an average of fifty-four months postoperatively. No acetabular component was revised because of aseptic loosening. According to Kaplan-Meier analysis, the seven-year survival rate, with an end point of femoral revision, osteolysis, or stem debonding, was 90.6 percent (95 percent confidence interval, 0.87 to 0.94). CONCLUSIONS: The prevalence of revision, osteolysis, and loosening after total hip arthroplasty with the Iowa femoral component at our institution was higher than that seen in our series of Harris Precoat stems, which had a survival rate of 98.4 percent (95 percent confidence interval, 0.97 to 1.00) at ten years with the same end points. The design of the Iowa stem may make it difficult to achieve a good cement mantle, and, in combination with the geometry and increased offset of the stem, may compromise the long-term survival of this cemented femoral component. PMID- 11005521 TI - The effect of femoral component head size on posterior dislocation of the artificial hip joint. AB - BACKGROUND: Posterior dislocation continues to be a relatively common complication following total hip arthroplasty. In addition to technical and patient-associated factors, prosthetic features have also been shown to influence stability of the artificial hip joint. In this study, a dynamic model of the artificial hip joint was used to examine the influence of the size of the head of the femoral component on the range of motion prior to impingement and posterior dislocation following total hip replacement. METHODS: Six fresh cadaveric specimens were dissected, and an uncemented total hip prosthesis was implanted in each. Each specimen was mounted in a mechanical testing machine and loaded with use of a system of seven cables attached to the femur and pelvis that simulated the action of the major muscle groups crossing the hip joint. The hip was taken through a range of motion similar to that experienced when rising from a seated position. The three-dimensional position of the femur at the points of impingement and dislocation was recorded electronically. The range of joint motion was tested with prosthetic femoral heads of four different diameters (twenty-two, twenty-six, twenty-eight, and thirty-two millimeters). RESULTS: Significant associations were noted between the femoral head size and the degree of flexion at dislocation in ten (p = 0.001), twenty (p < 0.001), and thirty (p = 0.003) degrees of adduction. Increasing the femoral head size from twenty-two to twenty-eight millimeters increased the range of flexion by an average of 5.6 degrees prior to impingement and by an average of 7.6 degrees prior to posterior dislocation; however, increasing the head size from twenty-eight to thirty-two millimeters did not lead to more significant improvement in the range of joint motion. The site of impingement prior to dislocation varied with the size of the femoral head. With a twenty-two-millimeter head, impingement occurred between the neck of the femoral prosthesis and the acetabular liner, whereas with a thirty two-millimeter head, impingement most frequently occurred between the osseous femur and the pelvis. CONCLUSIONS: With the particular prosthesis that was tested, increasing the diameter of the femoral head component increased the range of motion prior to impingement and dislocation, decreased the prevalence of prosthetic impingement, and increased the prevalence of osseous impingement. CLINICAL RELEVANCE: These results suggest that femoral heads with a twenty-eight millimeter diameter increase the range of motion after total hip replacement. This may be beneficial when additional factors compromising joint stability are encountered. PMID- 11005522 TI - Calcaneal osteosarcoma associated with Werner syndrome. A case report with mutation analysis. PMID- 11005523 TI - Repetitive stress injury: diagnosis or self-fulfilling prophecy? AB - * The vague definitions of so-called repetitive stress injuries are indicative of the fact that scientific studies have failed to show that repetitive motion causes injury. * Given the uncertainty about causation, work-related musculoskeletal disorders (WRMSDs) is a more readily accepted term to describe these phenomena. * There is little doubt that most ergonomic interventions increase comfort in the work environment, which is of great benefit to the worker. Many proponents of ergonomics assert that the elimination of certain risk factors related to force, repetition, and posture can prevent or even cure work related musculoskeletal disorders of the upper extremity. However, there is little scientific support for this position. * Undue reliance on ergonomics to treat musculoskeletal disorders, to the exclusion of proper diagnosis and attention to medical and health risk factors, can have adverse consequences for the patient. * Science rather than politics and public policy should determine what causes injury and disease. * The failure of numerous plaintiffs in litigation regarding repetitive stress injury due to use of computer keyboards is important because, when judges and lay jurors were presented with both sides of the issue, they rejected these claims in a forum (the judicial system) that traditionally compensates individuals bringing so-called mass-tort cases. PMID- 11005524 TI - Glucosamine and chondroitin were found to improve outcomes in patients with osteoarthritis. PMID- 11005525 TI - Manual physical therapy and exercise improved function in osteoarthritis of the knee. PMID- 11005526 TI - Tourniquet use during arthroscopy did not adversely affect patient outcomes. PMID- 11005527 TI - Passive flexion sign: a simple tool for diagnosis of anterior interosseous nerve injury in children. PMID- 11005528 TI - Residency training. AB - R. C. is an orthopaedic resident in a teaching program. At the orthopaedic clinic, he examines an elderly, otherwise healthy patient who requires a total hip replacement. He presents the patient to his covering attending physician, who agrees to supervise the joint replacement surgery. The resident discusses the surgery with the patient. The procedure, risks, goals, benefits, and alternatives are presented. The patient agrees to proceed with the surgery. The resident performs the surgical procedure with the attending physician's assistance. The surgery lasts forty minutes longer than the attending physician's usual surgical time, and the blood loss is 300 milliliters greater. Postoperative radiographs demonstrate a well positioned press-fit acetabular component and a cemented femoral component in 6 degrees of varus. PMID- 11005529 TI - Bone regrowth after partial amputation for diabetes. PMID- 11005530 TI - The orthopaedic forum. PMID- 11005531 TI - The orthopaedic forum. PMID- 11005532 TI - Effect of disability compensation on patient outcomes. PMID- 11005533 TI - Giant-cell tumors of long bones. PMID- 11005534 TI - Sample size and statistical power. PMID- 11005535 TI - Peer review. The millennium engima: more is less. PMID- 11005536 TI - Hematoma as a complication of enoxaparin use. PMID- 11005537 TI - Vesalius's inconstant structures. PMID- 11005538 TI - Missed posterior fracture-dislocation of the humeral head. PMID- 11005539 TI - Transglenoid suture technique. PMID- 11005540 TI - Low propensity of conventional antiepileptic drugs for interaction with felbamate against maximal electroshock-induced seizures in mice. AB - The objective of this study was to evaluate an interaction of the novel antiepileptic drug felbamate (2-phenyl-1,3-propanediol dicarbamate) with conventional antiepileptic drugs against maximal electroshock-induced convulsions in mice. Electroconvulsions were produced by means of an alternating current (ear clip electrodes, 0.2-s stimulus duration, tonic hindlimb extension taken as the endpoint). Adverse effects were evaluated in the chimney test (motor performance) and passive avoidance task (long-term memory). Brain and plasma levels of antiepileptic drugs were measured by immunofluorescence. Felbamate (7.5-30 mg/kg) significantly increased the electroconvulsive threshold. Felbamate at the subprotective dose of 5 mg/kg did not affect the anticonvulsive action of antiepileptics studied. On the other hand, this drug used at the lowest protective dose of 7.5mg/kg remained without effect upon the activity of valproate, carbamazepine or phenobarbital, but significantly potentiated the protective potential of diphenylhydantoin. No adverse effects were observed with combinations of felbamate with these antiepileptics. Neither brain nor free plasma levels of antiepileptic drugs were changed by felbamate. The results indicate that the combination of felbamate with conventional antiepileptic drugs is not promising from the experimental point of view. PMID- 11005541 TI - Nitric oxide synthase does not mediate neurotoxicity after an i.c.v. injection of streptozotocin in the rat. AB - In the present study we evaluated the possible role of nitric oxide (NO) in mediating neuronal damage in middle-aged rats after an i.c.v. injection of streptozotocin (STREP). An i.c.v. injection of STREP has been reported to decrease the central metabolism of glucose. This inhibition of the energy metabolism after STREP treatment might induce an excitotoxic mechanism, which may lead to the stimulation of NO synthase and, consequently to the synthesis of NO. On the other hand, STREP might induce oxidative stress directly by liberation of NO from its nitroso moiety. To investigate whether NO synthase is involved in a possible excitotoxic mechanism after STREP treatment, some of the rats treated with STREP (1.25 mg/ kg in 4 microl, bilaterally 2 microl/injection site) were also treated with the NO synthase inhibitor N-nitro-L-arginine methyl ester (L NAME, 20 mg/kg i.p. 10 min, 6, 24 and 96 h after STREP injection). To investigate whether NO liberated from STREP may be responsible for neurotoxic effects, one additional group of control rats received an i.c.v. injection of the NO donor sodium nitroprusside (SNP, 10 microg in 4 microl). We found that STREP affected the behavioral performances in the open field and two-way active avoidance task. In addition, immunostaining for glial fibrillary acidic protein, an indicator of reactive astroglial changes to neuronal damage, showed that this was mainly located in peri- and paraventricular regions of the third and lateral ventricles, like for instance in the septum, caudate putamen and hippocampus. L-NAME treatment had no protective effect on the behavioral impairments and neuronal damage of STREP-treated rats. This suggests that the neuronal damage of STREP may still be a result of the decrease in the central energy metabolism, but without the involvement of NO synthase. This was supported by measuring, using immunostaining, the NO-mediated cyclic GMP production by the enzyme soluble guanylyl cyclase in cortical slices, i.e. L-NAME did not prevent NO production after STREP administration in vitro. In addition, it was found that SNP liberated NO in vitro, whereas in vivo SNP administration did not lead to any behavioral and neuronal deficits at all. However, the present study cannot exclude the involvement of NO liberated from STREP in neuronal damage. PMID- 11005542 TI - Serotonin and sexual behavior in the male rabbit. AB - Sexual behavior was evaluated in sexually experienced male rabbits after the administration of different serotonergic drugs. The serotonin1A receptor agonist 8-OH-DPAT, 1 mg/kg, inhibited male rabbit sexual behavior when animals were tested 15 min after subcutaneous (SC) administration of this compound. Lower doses, 0.25 and 0.5mg/kg, were ineffective at a test 30 min after drug injection. Furthermore, 8-OH-DPAT, 0.25mg/kg, failed to revert the inhibitory effects upon sexual behavior produced by lidocaine application to the rabbit penis. Stimulation of 5-HT1B/2C receptors by TFMPP, at doses between 0.625 and 2.5 mg/kg, produced a drastic inhibition of sexual behavior when the drug was administered SC 30 min before behavioral observation. Doses below 5mg/kg were ineffective when given intraperitoneally 15 min before test. When the 5-HT1D/2C receptors were stimulated by the agonist mCPP a reduced number of mounts and ejaculations was observed after the SC administration of 1.25 and 2.5 mg/kg. Similarly, the mixed 5-HT agonist/antagonist lisuride reduced the percentage of rabbits displaying mounting behavior at doses of 0.25 and 0.5 mg/kg SC. All compounds tested produced a clear inhibition of male rabbit sexual behavior independently of the receptor subtype activated. These results are at variance with previous observations in rats where 8-OH-DPAT and lisuride produced a drastic facilitation of masculine coital behavior. Moreover, while the inhibition of male sexual behavior in rats produced by TFMPP and mCPP is associated with a disruption of the execution of this behavior, in rabbits these compounds reduced sexual motivation. These results indicate that the effects of serotonergic drugs on sexual behavior are species specific. PMID- 11005544 TI - P300 component of visual event-related potentials distinguishes patients with idiopathic parkinson's disease from patients with essential tremor. AB - A considerable rate of misdiagnosis has recently been reported in movement disorders. One of the most difficult clinical problems is the differentiation of Parkinson's disease and essential tremor. In this study, we have examined whether event-related potentials (ERPs) could aid in the solution of this differential diagnostic problem. Visual ERPs were obtained by using an "oddball" paradigm in 20 patients with idiopathic Parkinson's disease (IPD), 20 patients with essential tremor (ET), and 20 age-matched control subjects. We found that the P300 amplitudes were selectively reduced in the IPD group at the midline (Fz, Cz, Pz) electrode sites, whereas the ET group exhibited selectively elevated P300 amplitudes at the left parietal and frontal (F3, P3) recording sites. The patients with IPD, but not ET, showed a significant prolongation of the P300 latency at each electrode site. Similarly, only the IPD patients were significantly slower in responding to the target stimulus. The primary visual evoked potential (P100) did not distinguish among the IPD, ET, and control subjects. These results suggest that visual ERPs might provide a useful diagnostic method for differentiating IPD from ET. PMID- 11005543 TI - The neuroprotective effects of (-)deprenyl in the gerbil hippocampus following transient global ischemia. AB - (-)Deprenyl (selegeline) is a monoamine oxidase B (MAO-B) inhibitor, but it also exerts several effects independent of MAO-B inhibition. For example, it has been shown to improve neuronal survival in different neurodegenerative models. In the present study, we have tested whether (-)deprenyl attenuates the neuronal damage in the hippocampus that is induced in a model of transient global ischemia in gerbils. (-)Deprenyl was administered 1) at a low daily dose starting two weeks before occlusion, 2) at a single high dose administered 3h after occlusion, or 3) at a low daily dose for one or two weeks after occlusion. A nonsignificant trend of reduced neuronal damage in the hippocampal CA1 area was seen in all experimental groups treated with (-)deprenyl, regardless of the timing of treatment. The results together with previous evidence suggest that (-)deprenyl may protect CA1 neurons from ischemia-induced delayed death by several possible mechanisms, including the suppression of oxidative stress and apoptotic processes. PMID- 11005545 TI - Naturally occurring and experimentally induced beta-amyloid deposits in the brains of marmosets (Callithrix jacchus). AB - Cerebral beta-amyloid occurs in elderly animals of some species and in Alzheimer's disease. Previously, we injected 3 young marmosets intracerebrally with brain tissue from a patient with Alzheimer's disease. Six years later, when the monkeys were middle aged, we found moderate numbers of intracerebral plaques and cerebrovascular deposits containing beta-amyloid. We have re-examined these brains and those of 10 other marmosets injected with brain homogenate containing beta-amyloid, and have found some beta-amyloid in animals injected more than 4 years previously. We have found beta-amyloid in 4 of 26 elderly control marmosets, but not in 9 young to middle-aged control marmosets. The beta-amyloid found in middle aged marmosets injected with Alzheimer brain tissue was, therefore, not a consequence of their age. Deposits in large cerebral vessels in elderly marmosets, and in marmosets previously injected with brain tissue containing beta-amyloid, reacted with antibodies to Abeta and Abeta1-40; plaques and microvessel deposits reacted with antibodies to Abeta and Abeta1-42. PMID- 11005546 TI - Sustained improvements in patients with dementia of Alzheimer's type (DAT) 6 months after termination of Cerebrolysin therapy. AB - The present study is an extension of the work of Ruther et al. (1994). 101 patients suffering from DAT were evaluated 6 months after completion of a 4 week (5 days per week) therapy with either 30 ml Cerebrolysin or placebo. The significant and clinically relevant improvements in the global rating (CGI), clinical symptomatology (SCAG), cognitive performance (ZVT-G) as primary efficacy variables, as well as the improvements in the secondary efficacy variables activities of daily living (NAI) and wellbeing (Bf-S), achieved in the Cerebrolysin group after only 4 weeks of active therapy, were maintained to a large extent during the follow-up period. Although there was a moderate tendency in the drug group towards loss of improvement, the differences between baseline and follow-up examination, as well as the differences between the verum and the placebo group, clearly document a sustained improvement in patients treated with Cerebrolysin in the first 4 weeks of the study period. It can be speculated that relatively short treatment courses with Cerebrolysin in patients suffering from neurodegenerative dementia can lead to long term influence on disease progression, which is in accordance with the proposed neurotrophic - nerve growth factor like - mode of action. PMID- 11005547 TI - Salivary, total plasma and plasma free cortisol in panic disorder. AB - BACKGROUND: Research on basal HPA axis activity in patients with panic disorder showed inconsistent results. METHODS: Basal total plasma, plasma free and salivary cortisol levels were compared in patients with panic disorder (n = 47) and in healthy individuals (n = 23). Correlations between these fractions were calculated. RESULTS: All three basal cortisol fractions were significantly elevated in patients compared to controls. There were significant correlations between all three cortisol fractions. CONCLUSIONS: Nonsignificant differences between cortisol levels of patients and healthy controls in previous studies may have been due to inclusion of less severely ill patients or to small sample sizes (96 words). PMID- 11005548 TI - Vitamin D in schizophrenia, major depression and alcoholism. AB - 25-Hydroxyvitamin D3, 1,25-dihydroxyvitamin D3, calcium, phosphate and parathyreoidal hormone levels were assessed in 34 patients with schizophrenia (DSM-III-R, 44% female, mean age 38.9 +/- 2.1 years), 30 patients with alcohol addiction (16% female, mean age 48.7 +/- 2.2 years), 25 patients with major depression (56% female, mean age 57.6+/- years) and 31 healthy controls. Only 25 hydroxyvitamin D3 and 1,25-dihydroxvitamin D3 levels were significantly lower in all groups of psychiatric patients than in normal controls, but not phosphate, calcium and parathyreoidal hormone levels. Significant differences in the vitamin D levels could not be found between the three psychiatric groups. These findings do not support the idea that vitamin D is specifically involved in the pathophysiology of depression. The difference in patients as compared to the healthy controls might be related to a different social background resulting in differing habits e.g. of nutrition. PMID- 11005549 TI - Stability of CSF metabolites measured by proton NMR. AB - Analysis of cerebrospinal fluid (CSF) metabolites can provide data regarding CNS involvement in neurologic and psychiatric illness. However, there is lack of research into the effect of processing and storage of CSF specimens on the levels of metabolites analyzed. CSF specimens from 10 depressed patients were analyzed by proton NMR before and after 72 hours exposure to room temperature. No effect of exposure was found on myoinositol, glucose, acetate, and alanine CSF levels and there was a substantial decrease of citrate (>50%) and increase in lactate, glutamine, creatine, and creatinine levels. PMID- 11005550 TI - Adjuvant radiation after lymph node dissection for melanoma. PMID- 11005551 TI - Breast conservation without irradiation for invasive disease: legendary quest or definable reality? PMID- 11005552 TI - Is adjuvant radiotherapy necessary after positive lymph node dissection in head and neck melanomas? AB - INTRODUCTION: Postoperative radiotherapy (PR) has been recommended in patients with advanced head and neck melanomas to improve regional control. This study examined the incidence of cervical recurrence among patients who did not receive PR after surgical management of node-positive head and neck melanomas. METHODS: A computerized search of a database listing more than 10,000 patients with melanoma prospectively acquired between 1971 and 1998 identified 217 patients with pathologically positive nodes who had undergone regional lymph node dissection (RLND). Of these patients, 21 had received PR and 196 had not. RESULTS: Median follow-up after RLND was 20 months for nonsurvivors and 32 months for survivors. The overall incidence of cervical recurrence was 14% (27/196). The 5-year cervical recurrence-free survival rate was 83%. Five-year cervical recurrence free survival rates were 69% vs. 87% for patients with vs. without extranodal disease (P = .004), 96% vs. 81% for patients with nonpalpable vs. palpable nodes (P = .0761), and 82% vs. 91% for patients with one to three positive nodes vs. more than three positive nodes (P = .256). Multivariate analysis, which included the timing of nodal disease presentation and the effect of systemic adjuvant therapy, identified extranodal disease as the only independent predictor of cervical recurrence (P = .034). Cervical recurrence was significantly related to the subsequent occurrence of distant relapse. CONCLUSIONS: The low incidence of cervical recurrence after RLND in patients with node-positive head and neck melanomas does not justify the routine use of PR. The only subset of patients who may benefit from PR are those with extranodal disease. PMID- 11005553 TI - Factors associated with local breast cancer recurrence after lumpectomy alone: postmenopausal patients. AB - BACKGROUND: We have been following a cohort of patients who underwent a lumpectomy without receiving adjuvant radiotherapy or adjuvant systemic therapy. We now report the experience of a postmenopausal subgroup. METHODS: The postmenopausal subgroup included 244 patients accrued between 1977 and 1986 and followed up. The end point was ipsilateral local breast cancer recurrence. The factors studied were the patient's age in years; tumor size (in mm); nodal status (N-, Nx, N+); estrogen and progesterone receptor status (< 10, - 10 fmol/mg protein); presence or absence of lymphovascular/perineural invasion; presence or absence, and type, of DCIS (none, non-comedo, comedo); percentage of DCIS; histological grade (1,2,3); and nuclear grade (1,2,3). Univariate analyses consisted of Kaplan-Meier plots and the Wilcoxon (Peto-Prentice) test statistic; the multivariate analyses were step-wise Cox and log-normal regressions. RESULTS: The median follow-up of those patients still alive was 9.1 years, and the overall relapse rate was 24% (59/244). The univariate results indicated that the characteristics of smaller tumor size, negative nodes, positive ER status, and no lymphovascular or perineural invasion were associated with significantly (P <.05) lower relapse. From the multivariate analyses, the factors lymphovascular or perineural invasion, age, and amount of DCIS were all significantly associated with local relapse with both Cox and log-normal regressions. Additionally, there was weak evidence of an association between ER (P = .08 in the Cox regression and in the log-normal) and nodal status (P = .09 in the log-normal regression) with local relapse. We also are able to define a low-risk subgroup (N-, age -65, no comedo, ER positive, no emboli) with a crude 10-year local recurrence rate of 9%. CONCLUSION: With longer follow-up, and for postmenopausal patients, there continues to be support for the theory that local relapse is affected by the factors lymphovascular or perineural invasion, age, amount of DCIS, ER, and nodal status. A low risk subgroup has been identified. PMID- 11005554 TI - Symposium overview: estrogens and antiestrogens in managing the patient with breast cancer. AB - The prevalence of breast cancer (a hormonally driven neoplasm) in the United States, the potential health benefits of estrogen replacement therapy for postmenopausal women, and the burgeoning research focusing on selective estrogen receptor modulators (SERMs) have resulted in additional complexity in managing breast cancer. In an attempt to clarify existing data, the Society of Surgical Oncology sponsored a symposium entitled "Estrogens and Antiestrogens in Managing the Patient with Breast Cancer" at its 52nd Annual Cancer Symposium. This conference was held in March 1999 and was chaired by Dr. S. Eva Singletary, Professor of Surgery and Chief of the Surgical Breast Section at The University of Texas M. D. Anderson Cancer Center in Houston, Texas. The following is a review of the material presented by the symposium participants. PMID- 11005555 TI - Impact of sentinel lymph node mapping on relative charges in patients with early stage breast cancer. AB - BACKGROUND: The introduction of SLNB has allowed accurate staging in early-stage breast carcinomas and has minimized the number of unnecessary ALNDs. Intraoperative frozen-section analysis is a fundamental component of the sentinel lymph node biopsy (SLNB) procedure. Some patients have positive nodes on frozen section analysis and thus undergo a conventional axillary lymph node dissection (ALND) at the time of the SLNB. A few patients have negative nodes on frozen section analysis but have subsequent evidence of metastases on final pathologic examination. The purpose of our study was 2-fold: to compare the hospital-related charges of patients undergoing staging by SLNB with those of patients undergoing conventional ALND and to assess whether the different outcomes associated with SLNB adversely affect the charges incurred with this procedure. METHODS: Our study group consisted of 100 patients with T1 breast cancer and breast conservation therapy who underwent either SLNB or ALND from July 1, 1997, to June 30, 1998. We identified the first 50 consecutive patients to undergo SLNB during this period. We chose a similar cohort of 50 patients for ALND. Mean hospital related charges for the SLNB patients were categorized and compared with those for the ALND patients. RESULTS: Results for the two groups were analyzed using a two-sample Wilcoxon rank-sum test. Charges for the OR and hospital stay were less for the SLNB group (P < .05). Frozen-section analysis in the SLNB group contributed to the significant difference in charges for pathologic evaluation. Overall, the two groups showed no significant difference in total hospital related charges. CONCLUSIONS: When SLNB is used for T1 tumors, a small percentage of patients (10% in our study) will return to the operating room for an ALND. This small percentage does not increase the charges related to SLNB, however, as the reduced stay for most patients offsets this subgroup's contribution to the total hospital-related charges. Thus, in patients with clinical stage I breast cancer, SLNB does not cause significantly higher hospital-related charges compared with conventional ALND. PMID- 11005556 TI - Impact of multicentricity on clinical outcome in patients with T1-2, N0-1, M0 breast cancer. AB - BACKGROUND: The objective was to determine the impact of multicentric breast cancer on recurrence and survival and to evaluate the current tumor, node, metastasis staging system recommendations for multicentricity in the breast. METHODS: This study included 284 nonpregnant patients with T1-2, N0-1, M0 breast cancer, without previous cancer, who were treated by modified radical mastectomy followed by doxorubicin-based adjuvant chemotherapy. Clinical and pathological data were collected retrospectively and survival was calculated from the date of initial diagnosis using the Kaplan-Meier method. RESULTS: The median follow-up time was 8 years (range, 0.3-24.0), and the median age was 47 years (range, 23 76). The median clinical size of the index tumor was 2.5 cm. In 17% of patients, the clinical nodal status was N1. In 84% of patients, pathology of the index lesion was invasive ductal +/- in situ. Multicentric breast cancer was detected in 60 patients (21%): 30 patients with two lesions, 13 patients with three lesions, and 17 patients with four or more lesions. Locoregional recurrence, contralateral breast cancer, distant metastasis, and survival (disease-specific and disease-free) were similar in both groups of multicentric versus unicentric breast tumors. There was a significant difference between groups in estrogen receptor and axillary lymph node positivity, but these did not contribute significantly to outcome on multivariate analysis. CONCLUSIONS: Multicentricity does not increase the risk of poor outcomes in patients with early-stage breast cancer. This supports the current recommendations of the tumor, node, metastasis staging system that tumor size should be based on the diameter of the largest lesion in patients with multicentric breast cancer. PMID- 11005557 TI - A prospective phase II trial of ONYX-015 adenovirus and chemotherapy in recurrent squamous cell carcinoma of the head and neck (the Baylor experience). AB - BACKGROUND: The E1-b attenuated adenovirus, ONYX-015 (Onyx Pharmaceuticals, Richmond, CA), has demonstrated antitumoral activity in patients with recurrent squamous cell carcinoma of the head and neck. This study evaluated the effects of intratumoral ONYX-015 injection combined with systemic chemotherapy. METHODS: Inclusion criteria included: (1) recurrent squamous cell carcinoma of the head and neck, not surgically salvageable, (2) target tumor amenable to direct injection, and (3) no prior chemotherapy for recurrent disease. Patients received ONYX-015 (10(10) plaque-forming units) intratumorally for 5 days, cisplatin (80 mg/m2) on day 1, and 5-fluorouracil (800-1000 mg/m2) on days 1-5. This cycle was repeated every 3 weeks. Serial physical examination and computed tomography were used to assess tumor size and treatment response. RESULTS: Fourteen patients were enrolled, and nine patients were evaluable for response at the time of enrollment. The mean age of the evaluable patients was 60.8 years (range, 46-71 years). Mean maximum tumor diameter was 4.8 cm (range, 1.9-10.5 cm). Treatment related toxicity included nausea (n = 7, 77.8%), vomiting (n = 5, 55.6%), mucositis (n = 5, 55.6%), pain at the injection site (n = 5, 55.6%), constipation (n = 4, 44.4%), and fatigue (n = 4, 44.4%). Locoregional tumor control was obtained in all nine patients (100%) (mean observation time, 157 days). Complete clinical response was seen in three patients (33.3%), partial response was seen in three patients (33.3%), minor response was seen in one patient (11.1%), and two patients (22.2%) had stable disease. Median time to local progression of disease has not been reached (range, 35-356 days). CONCLUSIONS: ONYX-015 adenovirus plus systemic cisplatin and 5-fluorouracil provides antitumor activity and local tumor control in patients with recurrent squamous cell carcinoma of the head and neck. This novel treatment approach offers hope for patients with limited treatment alternatives and provides the foundation for a phase III clinical trial. PMID- 11005558 TI - Radiofrequency ablation of 231 unresectable hepatic tumors: indications, limitations, and complications. AB - BACKGROUND: Radiofrequency ablation (RFA) is increasingly used for the local destruction of unresectable hepatic malignancies. There is little information on its optimal approach or potential complications. METHODS: Since late 1997, we have undertaken 91 RFA procedures to ablate 231 unresectable primary or metastatic liver tumors in 84 patients. RFA was performed via celiotomy (n = 39), laparoscopy (n = 27), or a percutaneous approach (n = 25). Patients were followed with spiral computed tomographic (CT) scans at 1 to 2 weeks postprocedure and then every 3 months for 2 years. RESULTS: Intraoperative ultrasound (IOUS) detected intrahepatic disease not evident on the preoperative scans of 25 of 66 patients (38%) undergoing RFA via celiotomy or laparoscopy. In 38 of 84 patients (45%), RFA was combined with resection or cryosurgical ablation (CSA), or both. RFA was used to treat an average of 2.8 lesions per patient, and the median size of treated lesions was 2 cm (range, 0.3-9 cm). The average hospital stay was 3.6 days overall (1.8 days for percutaneous and laparoscopic cases). Ten patients underwent a second RFA procedure (sequential ablations) and, in one case, a third RFA procedure for large (one patient), progressive (seven patients), and/or recurrent (three patients) lesions. Seven (8%) patients had complications: one skin burn; one postoperative hemorrhage; two simple hepatic abscesses; one hepatic abscess associated with diaphragmatic heat necrosis following sequential percutaneous ablations of a large lesion; one postoperative myocardial infarction; and one liver failure. There were three deaths, one (1%) of which was directly related to the RFA procedure. Three of the complications, including one RFA-related death, occurred after percutaneous RFA. At a median follow-up of 9 months (range, 1-27 months), 15 patients (18%) had recurrences at an RFA site, and 36 patients (43%) remained clinically free of disease. CONCLUSIONS: Celiotomy or laparoscopic approaches are preferred for RFA because they allow IOUS, which may demonstrate occult hepatic disease. Operative RFA also allows concomitant resection, CSA, or placement of a hepatic artery infusion pump, and isolation of the liver from adjacent organs. Percutaneous RFA should be reserved for patients at high risk for anesthesia, those with recurrent or progressive lesions, and those with smaller lesions sufficiently isolated from adjacent organs. Complications may be minimized when these approaches are applied selectively. PMID- 11005559 TI - Lymph node micrometastases do not predict relapse in stage II colon cancer. AB - BACKGROUND: Over one third of patients with stage II colonic adenocarcinoma experience tumor recurrence. Because effective adjuvant therapy is now available, it is important to identify subsets of patients at higher risk for relapse who may benefit from early treatment. Immunohistochemistry has been used to detect microscopic metastases in histologically uninvolved mesenteric lymph nodes, but the prognostic significance of minimal nodal involvement has not been established. METHODS: Hematoxylin and eosin (H&E)-stained recuts of 900 mesenteric lymph nodes from 55 patients (range, 2-47; mean, 16.4 nodes per case) with resected pT3 or pT4, N0, M0 (TNM stage II) colonic adenocarcinomas were re examined for the presence of metastases and then stained immunohistochemically for keratin using the AE1:AE3 antibody. Twenty-seven patients did not experience recurrence of tumor within 5 years following resection (no evidence of disease [NED]); 28 patients relapsed during the same time frame. Lymph nodes from 10 patients having colonic resections for nonneoplastic disorders also were stained as controls. Keratin-positive cells and cell clusters were quantified in the lymph nodes, and comparisons were made between patients with and without tumor relapse. RESULTS: In the relapse group, four patients had positive nodes already identified on the H&E-stained recuts and had to be excluded from further analysis. Sixteen additional patients had keratin-positive cells; thus, 16 of 24 (67%) had micrometastases. In the NED group, one patient had a positive node on H&E staining and 22 additional patients had keratin-positive cells, so 22 of 26 (84%) patients had micrometastases. In the patients who had micrometastases, there was a mean of 3.5 and 4.6 positive nodes in the relapse and NED groups, respectively, and a mean of 11.3 and 12.4 keratin-positive cells or clusters in the relapse and NED groups, respectively. No keratin-positive cells were found in the 1 to 21 (mean, 9.1) nodes per case studied in the control patients. CONCLUSIONS: Micrometastases to histologically uninvolved mesenteric lymph nodes commonly are detected in patients with pT3 or pT4 colonic adenocarcinomas on recuts stained immunohistochemically for keratin. Nodal micrometastases detected by immunohistochemical staining are not useful for identifying stage II patients at higher risk for relapse. PMID- 11005560 TI - Influence of multimodality therapy on the cellular immunity of patients with esophageal cancer. AB - BACKGROUND: Cancer patients have often been reported to have impaired immune function, and the effect of treatment modalities, such as surgery, irradiation, and chemotherapy, in depressing patients' immunity has also been reported. In this investigation, the effect of treatment on the cellular immunity of esophageal cancer patients was evaluated. METHODS: Immunological parameters, such as natural killer (NK) activity and lymphocyte subsets in peripheral blood, were measured in 32 esophageal cancer patients on 5 occasions (on the day of admission, 2 days before surgery, and 1 week, 1 month, and 2 months after surgery). RESULTS: NK activity was greatly impaired shortly after the operation, and the percentages of lymphocytes as a whole, and CD8+, CD16+, and CD57+ lymphocytes were significantly decreased, on the other hand, a postoperative increase in the CD4+/CD8+ ratio was observed. No significant depression of immune function by postoperative irradiation was observed. CONCLUSIONS: The results of this study suggest that cellular immunity, especially cytotoxicity, shortly after esophagectomy may be greatly impaired by the surgical stress of esophagectomy and an added effect of chemotherapy. PMID- 11005561 TI - Preoperative plasma plasminogen activator inhibitor type-1 and serum C-reactive protein levels in patients with colorectal cancer. The RANX05 Colorectal Cancer Study Group. AB - BACKGROUND: Preoperative plasma plasminogen activator inhibitor-1 (PAI-1) is a prognostic variable in patients with colorectal cancer. It has been suggested, however, that plasma PAI-1 is a nonspecific prognostic parameter similar to the acute-phase reactant C-reactive protein (CRP). In the present study we analyzed the association between plasma PAI-1 and serum CRP in patients scheduled for elective resection of colorectal cancer. In addition, the prognostic value of PAI 1 and CRP was studied in this patient cohort. METHODS: PAI-1 and CRP were analyzed in citrated plasma and serum, respectively, obtained preoperatively from 594 patients. Patients who required preoperative blood transfusion received SAGM blood, in which soluble PAI-1 is not present. None of the patients received pre- or postoperative adjuvant chemotherapy, and all were followed in the outpatient clinic for at least 5 years or until death. The association of PAI-1 and CRP, respectively, with survival was tested using the median value of PAI-1 and the upper normal limit for CRP. Analyses were performed by inclusion of all patients, and in the subgroup of patients, who underwent curative resection. RESULTS: The median follow-up period was 6.8 (5.4-7.9) years. The median value of plasma PAI-1 was 35.8 ng/ml, and values greater than 94 nmol/L identified patients with increased CRP levels. Comparison of the molecules showed that PAI-1 was weakly correlated with CRP (r = .26; P <.0001). Both molecules showed a Dukes independent distribution. In univariate survival analyses high levels of PAI-1 were found associated with poor prognosis and low levels with good prognosis (P = .02, HR: 1.3). Similarly, high levels of CRP were found associated with poor prognosis and low levels with good prognosis (P <.0001, HR: 1.9). In a multivariate statistical analysis including Dukes classification, gender, age, tumor location, perioperative blood transfusion, PAI-1 and CRP, plasma PAI-1 was a dependent prognostic variable, while serum CRP (P <.0001; HR: 1.4; 95% CI: 1.3 1.5) was found to be a Dukes independent prognostic variable. Similar analyses, excluding patients with Dukes' D disease showed serum CRP to be an independent prognostic variable (P <.0001; HR: 1.3: 95% CI: 1.2-1.5). CONCLUSIONS: This study did not show a strong correlation between plasma PAI-1 and serum CRP in patients with colorectal cancer. Serum CRP was found to be a Dukes independent prognostic variable in this patient cohort, and was found to identify a subgroup of curatively resected patients at risk for short survival. PMID- 11005562 TI - Reliability of lymphoscintigraphy in indicating the number of sentinel nodes in melanoma patients. AB - BACKGROUND: This study was undertaken to establish the reliability of lymphoscintigraphy in indicating the number of sentinel nodes in patients with melanoma. METHODS: Lymphoscintigraphy was performed with dynamic imaging after injection of 60 MBq 99mTc-nanocolloid (1.6 mCi) and static imaging after 2 hours in 200 patients with clinically localized primary melanoma of the skin. The following day, sentinel nodes were retrieved with the blue dye technique and a gamma detection probe (Neoprobe 1000/1500). The discrepancies between the number of sentinel nodes indicated by lymphoscintigraphy and the actual number of sentinel nodes as established by the surgeon were evaluated. RESULTS: Lymphoscintigraphy showed drainage to 393 sentinel nodes in 255 lymphatic fields in 199 patients. In 48 lymphatic fields (19%) in 46 patients (23%), the number of sentinel nodes was different from the number that was visualized with scintigraphy. Additional sentinel nodes were found by the surgeon because a lymphatic vessel was not seen on the lymphoscintigraphy (43%), because a sentinel node was not visualized separately from other hot nodes or vessels or the injection site (36%), or because a sentinel node was blue and not hot (4%). Fewer sentinel nodes were found than suggested by scintigraphy because a lymphangioma was mistaken for a sentinel node (4%) or because a single elongated node was depicted as two hot spots (6%). CONCLUSIONS: Although lymphoscintigraphy is indispensable for lymphatic mapping, the predicted number of sentinel nodes is accurate in only 81% of lymph node fields. The limited discriminating power of the gamma camera is an important cause of discrepancies. PMID- 11005563 TI - Cell cycle and apoptosis. AB - In multicellular organisms, cell proliferation and death must be regulated to maintain tissue homeostasis. Many observations suggest that this regulation may be achieved, in part, by coupling the process of cell cycle progression and programmed cell death by using and controlling a shared set of factors. An argument in favor of a link between the cell cycle and apoptosis arises from the accumulated evidence that manipulation of the cell cycle may either prevent or induce an apoptotic response. This linkage has been recognized for tumor suppressor genes such as p53 and RB, the dominant oncogene, c-Myc, and several cyclin-dependent kinases (Cdks) and their regulators. These proteins that function in proliferative pathways may also act to sensitize cells to apoptosis. Indeed, unregulated cell proliferation can result in pathologic conditions including neoplasias if it is not countered by the appropriate cell death. Translating the knowledge gained by studying the connection between cell death and cell proliferation may aid in identifying novel therapies to circumvent disease progression or improve clinical outcome. PMID- 11005564 TI - Mutation and expression of the DCC gene in human lung cancer. AB - Chromosome 18q is frequently deleted in lung cancers, and a common region of 18q deletions was mapped to chromosome 18q21. Since the DCC candidate tumor suppressor gene has been mapped in this region, mutation and expression of the DCC gene were examined in 46 lung cancer cell lines, consisting of 14 small cell lung carcinomas (SCLCs) and 32 non-small cell lung carcinomas (NSCLCs), to elucidate the pathogenetic significance of DCC alterations in human lung carcinogenesis. A heterozygous missense mutation was detected in a NSCLC cell line, Ma26, while homozygous deletion was not detected in any of the cell lines. The DCC gene was expressed in 11 (24%) of the 46 cell lines, and the incidence of DCC expression was significantly higher in SCLCs (7/14, 50%) than in NSCLCs (4/32, 13%) (P = .01, Fisher's exact test). Therefore, genetic alterations of DCC are infrequent; however, the levels of DCC expression vary among lung cancer cells, in particular, between SCLCs and NSCLCs. The present result does not implicate DCC as a specific mutational target of 18q deletions in human lung cancer; however, it suggests that DCC is a potential target of inactivation by genetic defects including intron or promoter mutations and/or epigenetic alterations. The present result also suggests that DCC expression is associated with some properties of SCLCs, such as a neuroendocrine (NE) feature. PMID- 11005565 TI - Anti-VEGF antibody treatment of glioblastoma prolongs survival but results in increased vascular cooption. AB - Vascular endothelial growth factor (VEGF) is an important mediator of the intense angiogenesis which is characteristic of glioblastoma. While genetic manipulation of VEGF/VEGF receptor expression has previously been shown to inhibit glioblastoma growth, to date, no study has examined the efficacy of pharmacologic blockade of VEGF activity as a means to inhibit intracranial growth of human glioblastoma. Using intraperitoneal administration of a neutralizing anti-VEGF antibody, we demonstrate that inhibition of VEGF significantly prolongs survival in athymic rats inoculated in the basal ganglia with G55 human glioblastoma cells. Systemic anti-VEGF inhibition causes decreased tumor vascularity as well as a marked increase in tumor cell apoptosis in intracranial tumors. Although intracranial glioblastoma tumors grow more slowly as a consequence of anti-VEGF treatment, the histologic pattern of growth suggests that these tumors adapt to inhibition of angiogenesis by increased infiltration and cooption of the host vasculature. PMID- 11005566 TI - Solitary lung tumors and their spontaneous metastasis in athymic nude mice orthotopically implanted with human non-small cell lung cancer. AB - We examined the tumorigenic and metastatic potentials of three human non-small cell lung cancer (NSCLC) cell lines, PC-14, A549 or Lu-99 cell lines suspended in Matrigel-containing phosphate-buffered saline were orthotopically implanted into the lungs of nude mice. The formation of a solitary tumor nodule in the lung was observed after the implantation of all cell lines. Intrapulmonary implantation of PC-14 or Lu-99 cells resulted in spontaneous distant metastases. In contrast, A549 cells caused multiple intrapulmonary metastases to the right and left lobes of the lung without producing visible lymphatic metastasis. We also investigated the expression of matrix metalloproteinases (MMPs), urokinase-type plasminogen activator (u-PA), u-PA receptor (u-PAR) and c-MET in these cell lines in vitro and in vivo. Reverse transcription polymerase chain reaction (RT-PCR) analysis showed that the expression of MMP-2 and membrane-type 1 MMP (MT1-MMP) was elevated in PC-14 as compared with the other two cell lines. In contrast, stronger expression of c-MET was observed in A549 than in PC-14 or Lu-99. These results indicate that differential patterns of metastasis of lung cancer might be associated with differential expression of metastasis-associated molecules. Our orthotopic implantation models display clinical features resembling those of NSCLC, and may provide a useful basis for lung cancer research. PMID- 11005567 TI - A mathematical model for comparison of bolus injection, continuous infusion, and liposomal delivery of doxorubicin to tumor cells. AB - Determining the optimal mode of delivery for doxorubicin is important given the wide use of the drug against many tumor types. The relative performances of bolus injection, continuous infusion, liposomal and thermoliposomal delivery are not yet definitely established from clinical trials. Here, a mathematical model is used to compare bolus injection, continuous infusion for various durations, liposomal and thermoliposomal delivery of doxorubicin. Effects of the relatively slow rate, and saturability, of doxorubicin uptake by cells are included. Peak concentrations attained in tumor cells are predicted and used as a measure of antitumor effectiveness. To measure toxicity, plasma area under the curve (AUC) and peak plasma concentrations of free doxorubicin are computed. For continuous infusion, the duration of infusion significantly affects predicted outcome. The optimal infusion duration increases with dose, and is in the range 1 to 3 hours at typical doses. The simulations suggest that continuous infusion for optimal durations is superior to the other protocols. Nonthermosensitive liposomes approach the efficacy of continuous infusion only if they release drug at optimal rates. Predictions for thermosensitive liposomes indicate a potential advantage at some doses, but only if hyperthermia is applied locally so that the blood is not significantly heated. PMID- 11005568 TI - Caspase-dependent apoptosis induced by telomere cleavage and TRF2 loss. AB - Chromosomal abnormalities involving telomeric associations (TAs) often precede replicative senescence and abnormal chromosome configurations. We report here that telomere cleavage following exposure to proapoptotic agents is an early event in apoptosis. Exposure of human and murine cancer cells to a variety of pro apoptotic stimuli (staurosporine, thapsigargin, anti-Fas antibody, and cancer chemotherapeutic agents) resulted in telomere cleavage and aggregation, and finally their extrusion from the nuclei. Telomere loss was associated with arrest of cells in G2/M phase and preceded DNA fragmentation. Telomere erosion and subsequent large-scale chromatin cleavage were inhibited by overexpression of the anti-apoptotic protein, bcl-2, and two peptide caspase inhibitors (BACMK and zVADfmk), indicating that both events are regulated by caspase activation. The results demonstrate that telomere cleavage is an early chromatin alteration detected in various cancer cell lines leading to drug-induced apoptosis, and suggest that this event contributes to mitotic catastrophe and induction of cell death. Results also suggest that the decrease of telomeric-repeat binding factor 2 (TRF2) may be the earliest event in the ara-C-induced telomere shortening, induction of endoreduplication and chromosomal fragmentation leading to cell death. PMID- 11005569 TI - Indomethacin-induced apoptosis in esophageal adenocarcinoma cells involves upregulation of Bax and translocation of mitochondrial cytochrome C independent of COX-2 expression. AB - The prolonged use of nonsteroidal anti-inflammatory drugs (NSAIDs) has been shown to exert a chemopreventive effect in esophageal and other gastrointestinal tumors. The precise mechanism by which this occurs, however, is unknown. While the inhibition of COX-2 as a potential explanation for this chemopreventive effect has gained a great deal of support, there also exists evidence supporting the presence of cyclooxygenase-independent pathways through which NSAIDs may exert their effects. In this study, immunohistochemical analysis of 29 Barrett's epithelial samples and 60 esophageal adenocarcinomas demonstrated abundant expression of the COX-2 protein in Barrett's epithelium, but marked heterogeneity of expression in esophageal adenocarcinomas. The three esophageal adenocarcinoma cell lines, Flo-1, Bic-1, and Seg-1, also demonstrated varying expression patterns for COX-1 and COX-2. Indomethacin induced apoptosis in all three cell lines, however, in both a time- and dose-dependent manner. In Flo-1 cells, which expressed almost undetectable levels of COX-1 and COX-2, and in Seg-1, which expressed significant levels of COX-1 and COX-2, indomethacin caused upregulation of the pro-apoptotic protein Bax. The upregulation of Bax was accompanied by the translocation of mitochondrial cytochrome c to the cytoplasm, and activation of caspase 9. Pre-treatment of both cell lines with the specific caspase 9 inhibitor, z-LEHD-FMK, as well as the broad-spectrum caspase inhibitor, z-VAD FMK, blocked the effect of indomethacin-induced apoptosis. These data demonstrate that induction of apoptosis by indomethacin in esophageal adenocarcinoma cells is associated with the upregulation of Bax expression and mitochondrial cytochrome c translocation, and does not correlate with the expression of COX-2. This may have important implications for identifying new therapeutic targets in this deadly disease. PMID- 11005572 TI - Polymer-supported and chemoenzymatic synthesis of the Neisseria meningitidis pentasaccharide: a methodological comparison. AB - Neisseria meningitidis trisaccharide [GlcNAc[(1-->3)Galbeta(1-->4)Glc-R], tetrasaccharide [Galbeta(1-->4)GlcNAcbeta(1--> 3)Galbeta(1-->4)Glc-R], and a pentasaccharide [Neu5Acalpha(2-->3)Galbeta(1-->4)GlcNAcbeta(1-->3)G albeta(1- >4)Glc-SPh] were prepared via conventional chemical synthesis, polymer-supported synthesis, and chemoenzymatic methods, starting from D-lactose. The polymer polyethyleneglycol monomethylether (MPEG) and the linker dioxyxylene (DOX) were used with a lactose-bound acceptor to improve the purification process. Several enzymes (LgtA, GalE-LgtB fusion, and CMP-Neu5Ac synthetase/sialyltransferase fusion) were used for syntheses of these oligosaccharides. Excellent stereo- and regioselectivities as well as high yield (> 90% from Gal(1-->4)Glc-SPh) of the pentasaccharide were obtained. Both of the convenient processes are suitable for efficient preparation of target oligosaccharides. PMID- 11005570 TI - Ras-mediated suppression of TGFbetaRII expression in intestinal epithelial cells involves Raf-independent signaling. AB - Ras-transformed intestinal epithelial cells are resistant to the growth inhibitory actions of TGFbeta and have a marked decrease in expression of the TGFbeta type II receptor (TGFbetaRII). Rat intestinal epithelial cells (RIE) were stably transfected with activated Ras, Sos and Raf constructs and tested for expression of TGFbetaRII and sensitivity to growth inhibition by TGFbeta. The parental RIE line and the RIE-Raf cells were non-transformed in morphology and were sensitive to TGFbeta (70-90% inhibited). In contrast, the RIE-Ras and RIE Sos lines were transformed, resistant to TGFbeta and expressed 5- to 10-fold decreased levels of the TGFbetaRII mRNA and protein. Cyclin D1 protein expression was repressed by TGFbeta treatment in parental RIE and RIE-Raf cells, whereas levels of cyclin D1 in RIE-Ras and RIE-Sos cells remained unchanged. Treatment of RIE-Ras cells with 25 microM farnesyl transferase inhibitor, FTI L739,749, for 48 hours restored expression of TGFbetaRII to levels equivalent to control cells. In addition, treatment of RIE-Ras cells for 48 hours with PD-98059, a specific MAPKK inhibitor, also increased expression of TGFbetaRII to control levels. Collectively these results suggest that downregulation of TGFbetaRII and loss of sensitivity to growth inhibition by TGFbeta in Ras-transformed intestinal epithelial cells is not mediated exclusively by the conventional Ras/Raf/MAPKK/MAPK pathway. However, activation of MAPK, perhaps by an alternate Ras effector pathway, appears to be necessary for Ras-mediated downregulation of TGFbetaRII. PMID- 11005571 TI - HGF/SF activates glycolysis and oxidative phosphorylation in DA3 murine mammary cancer cells. AB - Hepatocyte growth factor/scatter factor (HGF/SF) is a paracrine growth factor which increases cellular motility and has also been implicated in tumor development and progression and in angiogenesis. Little is known about the metabolic alteration induced in cells following Met-HGF/SF signal transduction. The hypothesis that HGF/SF alters the energy metabolism of cancer cells was investigated in perfused DA3 murine mammary cancer cells by nuclear magnetic resonance (NMR) spectroscopy, oxygen and glucose consumption assays and confocal laser scanning microscopy (CLSM). 31P NMR demonstrated that HGF/SF induced remarkable alterations in phospholipid metabolites, and enhanced the rate of glucose phosphorylation (P < .05). 13C NMR measurements, using [13C1]-glucose enriched medium, showed that HGS/SF reduced the steady state levels of glucose and elevated those of lactate (P < .05). In addition, HGF/SF treatment increased oxygen consumption from 0.58+/-0.02 to 0.71+/-0.03 micromol/hour per milligram protein (P < .05). However, it decreased CO2 levels, and attenuated pH decrease. The mechanisms of these unexpected effects were delineated by CLSM, using NAD(P)H fluorescence measurements, which showed that HGF/SF increased the oxidation of the mitochondrial NAD system. We propose that concomitant with induction of ruffling, HGF/SF enhances both the glycolytic and oxidative phosphorylation pathways of energy production. PMID- 11005573 TI - Syntheses of four D- and L-hexoses via diastereoselective and enantioselective dihydroxylation reactions. AB - An expeditious approach to various protected hexoses has been developed by the use of the Sharpless catalytic asymmetric dihydroxylation reaction. Applying the Sharpless catalytic asymmetric dihydroxylation reaction on vinylfuran, diols with high enantioexcess are produced. The resulting diols can be stereoselectively transformed into either protected D- or L-mannose in five steps and approximately 39% yield from furfural. Similarly, both D- and L-talose and gulose have been synthesized in 19% overall yields, respectively. Using a modified strategy, both protected D- and L-gulo- and allo-sugar-delta-lactones were synthesized in eight steps and approximately 20%, overall yield from furfural. PMID- 11005574 TI - Synthesis of novel 3'-C-methyl-apionucleosides: an asymmetric construction of a quaternary carbon by Claisen rearrangement. AB - The synthesis of 2,3-dideoxy-3-C-(hydroxymethyl)-3-C-methyl-D-glycero tetrofuranosyl++ + nucleosides was accomplished in high enatiomeric purity (98.5% ee) via [3,3]-sigmatropic Claisen rearrangement of (E)(S)-5-benzyloxy-1-tert butyldimethylsilanyloxy-4-methyl-pent-3- en-2-ol prepared from 2,3-O isopropylidene-D-glyceraldehyde. The synthesized nucleosides were assayed against human immunodeficiency virus (HIV) and hepatitis B virus in human peripheral blood mononuclear (PBM) and 2.2.15 cells, respectively. 6-Amino-9-[2,3-dideoxy-3 C-(hydroxymethyl)-3-C-methyl-beta-D-glycero- tetrofuranosyl]-2-fluoropurine shows moderate antiviral activity (EC50 = 2.55 microM) against HIV-1 strains and 6 amino-9-[3-deoxy-3-C-(hydroxymethyl)-3-methyl-alpha-D-glycero-tetro furanosyl]-2 fluoropurine exhibits potent anti-HIV activity (EC50 = 0.073 microM) with significant cytotoxicity (IC50 = 1.0 microM). PMID- 11005575 TI - Synthesis of 2',3'-dideoxy-3'-fluoro-L-ribonucleosides as potential antiviral agents from D-sorbitol. AB - 2',3'-Dideoxy-3'-fluoro-L-ribonucleosides were synthesized as potential antiviral agents. The key intermediate, methyl 5-O-benzoyl-2,3-dideoxy-3-fluoro-L ribofuranoside, which was prepared from D-sorbitol, was condensed with pyrimidine and purine bases to obtain the respective nucleosides. Among them, the cytosine analogue 2',3'-dideoxy-3'-fluoro-alpha-L-cytidine showed a moderate anti-HBV activity. PMID- 11005576 TI - Selective formation of C-2 azidodeoxy-D-glucose derivatives from D-glucal precursors using the azidonitration reaction. AB - A series of glucals, protected by cyclic acetal protecting groups to conformationally constrain the C-4 and C-6 hydroxyl groups, were subjected to the azidonitration reaction to furnish the corresponding C-2 azidodeoxy-D-glucoses. 4,6-O-Isopropylidene-3-O-triisopropylsilyl-D-arabino-hex-1-enit ol afforded 2 azido-2-deoxy-4,6-O-isopropylidene-3-O-triisopropylsilyl-D-gluco pyranosyl nitrate and its D-manno isomer in a 20:1 ratio. These findings allow the azidonitration reaction to be now used for the preparation of a variety of glucosamine building blocks from differentially protected glucal precursors. PMID- 11005577 TI - Synthesis of the HSA-conjugate of the S-linked thiomimetic of the branched tetrasaccharide repeating unit of the immunostimulant polysaccharide, schizophyllan. Evaluation as potential immunomodulator. AB - The conjugate with human serum albumin (HSA) of the S-linked thioanalogue of the branched tetrasaccharide repeating unit of the polysaccharide, schizophyllan, was synthesized from 1,2,4,6-tetra-O-acetyl-3-S-[2,4-di-O-acetyl-3,6-di-S-(2,3,4,6 tetra-O-ac etyl-beta-D-glucopyranosyl)-3,6-dithio-beta-D-glucopyranosyl]-3-thio bet a-D-glucopyranose [M.O. Contour-Galcera et al., Carbohydr. Res., 281 (1996) 119-128] in five steps, and its potential immunomodulatory activity was evaluated in human blood mononuclear cells. The protein glycoconjugate did not effect proliferation or production of IL-4, IL-5 and IFN-g in a significant way. PMID- 11005578 TI - The use of the 2-aminobenzoic acid tag for oligosaccharide gel electrophoresis. AB - Gel electrophoresis of fluorophore labeled saccharides provides a rapid and reliable method to screen enzymatic and/or chemical treatments of polysaccharides and glycoconjugates, as well as a sensitive and efficient microscale method to separate and purify oligosaccharides for further analysis. A simple and inexpensive method of derivatization and analysis using 2-aminobenzoic acid (anthranilic acid, AA) is described and applied to the extracellular polysaccharide released by the desiccation tolerant cyanobacterium Nostoc commune DRH-1. The results of these analyses suggest a possible protective functionality of two pendent groups, as well as a potential relationship between these groups and the desiccation tolerance of the organism. PMID- 11005579 TI - Ischemic preconditioning. PMID- 11005580 TI - Do mitochondrial K(ATP) channels serve as triggers rather than end-effectors of ischemic preconditioning's protection? PMID- 11005581 TI - Opening mitochondrial K(ATP) in the heart--what happens, and what does not happen. AB - There is considerable evidence that opening the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) is cardioprotective in ischemia-reperfusion. Two prominent questions surround the role of mitoK(ATP) in the cardiomyocyte: How does opening mitoK(ATP) protect? What is the normal physiological role of mitoK(ATP) in the heart? Before these questions can be addressed, it is necessary to agree on the bioenergetic consequences of opening mitoK(ATP), and this distills down to a single question--does opening mitoK(ATP) cause significant uncoupling or not? The evidence strongly indicates that it does not and that reports of uncoupling and inhibition of Ca2+ uptake are the result of using toxic concentrations of K(ATP) channel openers. Thus, opening mitoK(ATP) results in increased K+ flux that is sufficient to change mitochondrial volume but is insufficient to cause significant depolarization of membrane potential. The volume changes, however, have significant bioenergetic consequences for energy coupling in the cell. PMID- 11005582 TI - The role of mitochondrial KATP channels in cardioprotection. PMID- 11005583 TI - The role of mitochondrial K(ATP) channels in cardioprotection. PMID- 11005584 TI - Potential role of endothelin-1 and endothelin antagonists in cardiovascular diseases. AB - The endothelins comprise a family of three isopeptides ET-1, ET-2 and ET-3, whereby ET-1 appears to be the most relevant in humans. They act in a paracrine manner on ETA and ETB receptors. ET-1 plays an important role in the cardiovascular system. In addition, it modulates vasomotion and growth processes, and it participates in thrombogenesis and neutrophil adhesion. This review summarizes some of the current literature pertaining to the physiological and pathophysiological significance of ET-1, focusing the assets and drawbacks of elevated ET-1 levels. In this regard, modulation of the endothelin system by either receptor blockade or by inhibition of endothelin converting enzyme is expected to provide novel therapeutic drug strategies. PMID- 11005585 TI - Influence of intercellular junctions on endothelin secretion of human umbilical vein endothelial cells in vitro. AB - The endothelium plays a pivotal role in the rheological regulation of blood flow by the secretion of vasoactive factors. The interaction between shear forces and the endothelium is determined by the mechanical properties of the endothelial cell layer which are associated with intercellular junctions. Cell-cell contacts could therefore modulate the secretion of vasocative factors in response to rheological stimuli. We investigated the relationship between intercellular junctions and the secretion of the vasoconstrictor peptide endothelin and the coagulation co-factor von Willebrand factor (vWF). Human umbilical vein endothelial cells (HUVECs) were used as in vitro endothelial model system. Intercellular junctions were reversibly disrupted by calcium chelation or hypertonic stress; alternatively, the formation of intercellular junctions was inhibited by culturing the cells in suspension or by plating them in the presence of an inhibitory anti-VE-cadherin antibody. The opening of intercellular junctions was verified by assessing transmonolayer electrical resistance (TMR) and immunofluorescence morphology. The concentration of endothelin and vWF was measured in the cell culture supernatants using specific ELISAs. The secretion of endothelin was inhibited by EGTA (5 mM) and stimulated by incubation with tumor necrosis factor alpha (TNFalpha, 40 ng/ml). Treatment with hypertonic medium (glycerol, 1,200 mosmol/l) for 10 minutes opened intercellular junctions and markedly reduced the secretion of endothelin. HUVECs in suspension culture did not secrete endothelin and failed to respond to TNFalpha, but readily resumed these functions upon forming a new monolayer on plastic. The reconstitution of intercellular junctions after suspension culture could be inhibited using a specific anti-VE-cadherin antibody. This antibody, but not a non-specific anti human-IgG antibody reduced endothelin secretion. The secretion of von Willebrand Factor was less dependent on intercellular junctions. The opening of intercellular junctions did not induce cell death, since the cells continued to exclude trypan blue. The results of this study suggest a novel and potentially pathophysiologically/clinically relevant correlation between intercellular junctions and the secretion of endothelin in endothelial cells. PMID- 11005586 TI - Lack of nuclear apoptosis in cardiomyocytes and increased endothelin-1 levels in a rat heart model of myocardial stunning. AB - OBJECTIVE: Reperfusion injury may affect the cardiac NO and endothelin production. We investigated whether 20 min of total ischemia followed by 40 min of reperfusion can induce apoptosis in a Langendorff model of retrogradely perfused rat hearts (37 degrees C; paced at 300/'), and we attempted to correlate these findings with measured tissue NO and ET-1 levels. METHODS: An apoptosis detection system was utilized which catalytically incorporates fluorescein-12 dUTP at the 3'-OH DNA ends using the principle of the TUNEL assay, with direct visualization of the labeled DNA. ET-1 was measured by radioimmunoassay and NO3/NO2 by ion pairing HPLC on C18 reverse phase columns. RESULTS: None of the postischemic (n = 6) nor of the control perfused (90 min, n = 6) hearts showed signs of apoptosis, while those exposed to longer ischemia (40 min) and reperfusion (2 h) confirmed the presence of apoptotic cells. Myocardial ET-1 concentrations were 4.8 +/- 1.0 versus 8.3 +/- 2.5 pg/100 mg (control vs. ischemic hearts, respectively; mean +/- SD; p < 0.05). Myocardial NO contents showed no differences. CONCLUSION: These data suggest that the time window of apoptosis with detectable DNA fragmentation exceeds 20 min of global total ischemia and 40 min of reperfusion, a model frequently used for inducing myocardial stunning. While NO was not increased in postischemic hearts, increased ET-1 levels indirectly argue for a role of ET-1 as inducer of apoptosis, but only at a later stage of reperfusion. PMID- 11005587 TI - Differences in time course of myocardial mRNA expression in non-infarcted myocardium after myocardial infarction. AB - In non-infarcted myocardium after myocardial infarction, the change of cardiac phenotypic modulation of contractile protein, extracellular matrix and intracellular Ca2+ transport protein, such as sarcoplasmic reticulum Ca2+(SR Ca2+)-ATPase, Na+-Ca2+ exchanger, have a important role during cardiac remodeling. However, the time course in this gene expression in the adjacent and remote left ventricular, or right ventricular myocardium after myocardial infarction has not been well examined. The purpose of this study was to examine the left ventricular function and regional cardiac gene expression after myocardial infarction. Myocardial infarction was produced in Wistar rats by the ligation of the left anterior descending coronary artery. After 3 weeks, 2 months and 4 months from myocardial infarction, we performed Doppler echocardiography and measured the systolic and diastolic function. Then, we analyzed the contractile protein, extracellular matrix and intracellular Ca2+ transport protein mRNAs of cardiac tissues in the adjacent and the remote noninfarcted myocardium, and right ventricular myocardium by Northern blot hybridization. Fractional shortening of infarcted heart progressively decreased. Peak early diastolic filling wave (E wave) velocity increased, and the deceleration rate of the E wave velocity was more rapid in myocardial infarction areas. Atrial filling wave (A wave) velocity decreased, resulting in a marked increase in the ratio of E wave to A wave velocity. Expression of myocardial alpha-skeletal actin, beta MHC and ANP mRNA, or collagen I and III mRNA were higher at 3 weeks after myocardial infarction. SR Ca2+-ATPase mRNA in the adjacent non-infarcted myocardium was decreased at 2 months, and that in remote myocardium was decreased at 4 months after infarction. Na+-Ca2+ exchanger mRNA levels were increased at 3 weeks, but was decreased at 2 months in the adjacent non-infarcted myocardium and at 4 months in the remote myocardium. These findings suggest that the compensation for myocardial infarction by myocardial gene expression in non infarcted myocardium may occur at an early phase after myocardial infarction, and myocardial dysfunction may begin from adjacent to remote non-infarcted myocardium during progressive cardiac remodeling. PMID- 11005588 TI - Electrophysiologic effects of the new I(Ks)-blocking agent chromanol 293b in the postinfarction canine heart. Preserved positive use-dependence and preferential prolongation of refractoriness in the infarct zone. AB - OBJECTIVES: Aim of the present study was to investigate site- and rate dependent effects of the IKs-blocking agent chromanol 293b on conduction and refractoriness in normal, infarcted, and transitional regions of in-situ canine hearts. METHODS: In five dogs with subacute myocardial infarction, three-dimensional mapping was performed after insertion of 6 x 6 needle electrodes in the left ventricle. Before and after application of chromanol 293b (10 mg/kg), activation patterns and local refractory periods (ERPs) at pacing intervals of 300, 500 and 850 ms were obtained for the surviving epicardial muscle layer of the infarct zone (IZ) and for epi-, endo-, and midmyocardial muscle layers of both the normal zone (NZ) and the border zone (BZ) separating normal and infarcted areas. RESULTS: At baseline, both the NZ and the BZ exhibited uniform ERPs throughout the ventricular wall. Epicardial ERPs were longer in the IZ than in the NZ, and intermediate in the BZ. Chromanol 293b did not affect total activation times. However, at fast heart rates regional areas of slow conduction occurred. Chromanol 293b ubiquitously prolonged local ERPs, most markedly in the IZ. A preferential effect on individual muscle layers of the NZ or BZ and, thus, drug induced transmural dispersion of ERP could not be observed. Again ubiquitously, the effect on ERP was more pronounced at faster than at slower heart rates, that is, positive use-dependent. At a basic cycle length of 300 ms, chromanol 293b prolonged local ERPs in the IZ by 46 +/- 24 %, in the BZ by 34 +/- 26%, and in the NZ by 20 +/- 17% (p < or = 0.05). CONCLUSIONS: At least in theory, the electrophysiologic properties of chromanol 293 b, that is, preferential prolongation of refractoriness in ischemic myocardium, more pronounced at faster than at slower heart rates, but homogeneously throughout the intact ventricular wall, appear to be favorable. Whether this translates into a clinical benefit, particularly in the treatment of ischemia-related ventricular tachyarrhythmias, remains to be determined. PMID- 11005589 TI - Inefficient ventilation and reduced respiratory muscle capacity in congestive heart failure. AB - The extent and time-course of changes in lung volumes, ventilatory efficiency at rest and during exercise, and respiratory muscle function and their influence on exercise limitation in congestive heart failure (CHF) are unclear. It is unknown whether respiratory muscle function may predict changes in exercise limitation or may be impaired in patients with poor outcome. 145 male patients (54 +/- 1 years) suffering from CHF (NYHA class I-III, mean 2.3 +/- 0.1), with a LVEF of 23 +/- 1%, and a mean peak O2 uptake (VO2peak) 15.0 +/- 0.5 mL X min(-1) X kg(-1), were studied. They were grouped in Weber functional classes A to D according to their VO2peak. Significant increases in ventilatory equivalents for O2 and CO2 (VE/VCO2peak) and in dead space ventilation at rest and during exercise were found with increasing exercise limitation. Moreover, there was a correlation of static and dynamic lung volumes (inspiratory vital capacity, IVC, r = 0.43, P < 0.01), as well as of maximal inspiratory pressure (MIP; r = 0.46, P < 0.01) with VO2peak. Patients who died (n = 26) or were heart transplanted (n = 20) during a follow-up (mean 800 +/- 10 days) had a reduced MIP (6.4 +/- 0.4 kPa) as compared with survivors (n = 82; 9.3 +/- 0.7 kPa, P < 0.01). In a subgroup of 33 patients re-evaluated after six months, individual changes in IVC and VE/CO2peak, but not in MIP, correlated to changes in VO2peak (r = 0.69 and r = 0.72 respectively; P < 0.01). In CHF, exercise limitation is associated with reversible lung restriction and inefficient ventilation at rest and during exercise. Patients with severe CHF have a significant reduction in MIP, a finding that is associated with poor outcome. PMID- 11005590 TI - Robotically enhanced "Dresden technique" with bilateral internal mammary artery grafting. AB - BACKGROUND: A tendency to reduce the operative trauma seems to be the prominent sign of development in cardiac surgery. The combination of the previously introduced minimally invasive "Dresden Technique" with the robotically enhanced endoscopic system enables further progress in coronary artery surgery. METHODS: From May 1999, 25 patients (19 male, 6 female, mean age 63 +/- 8.0 years) suffering from multivessel coronary artery disease (CAD) were treated surgically using arterial revascularization through the bilateral internal mammary arteries (BIMA). Both mammaries were harvested endoscopically using the da Vinci robotic system. Further, the "Dresden Technique" was applied for anastomosis of these vessels with the coronaries. RESULTS: All patients survived the surgery. Mean duration of surgery amounted (244 +/- 82.1 min). BIMA dissection took (87 +/- 18.1 min). An average of 2.2 anastomoses were performed per patient. Postoperatively, the patients remained on ICU for 19 +/- 18 hrs. One patient needed reexploration for bleeding (4%). CONCLUSIONS: The addition of robotically enhanced endoscopic mammary artery harvesting method to the "Dresden Technique" for coronary artery surgery enables the introduction of a new treatment method for CAD into surgical practice, helping to perform an arterial revascularization with a distinctly reduced surgical trauma. PMID- 11005591 TI - Robotic coronary artery surgery--the evolution of a new minimally invasive approach in coronary artery surgery. AB - BACKGROUND: With the introduction of a wrist-enhanced robotic surgical system into minimally invasive cardiac surgery, the outlook of performing closed chest coronary artery bypass operations became a reality. METHODS: Since May, 1999, this new wrist-enhanced instrumentation has been used in 109 (79 male, 30 female, median age 63 +/- 9.9 years) patients. Seven suffering from single vessel coronary artery disease (SVD) and double vessel disease (DVD) were treated as totally endoscopic coronary artery bypass (TECAB). 60 with SVD underwent a wrist enhanced, minimally invasive direct coronary artery bypass procedure. 25 with DVD were treated using the robotically enhanced Dresden Technique coronary artery bypass. RESULTS: Survival was 100%. In all patients the IMAs were safely harvested totally endoscopically. In the TECAB group, all patients were operated via a three-point stab incision. Data were observed during and after the operation. CONCLUSIONS: Our preliminary experiences with this new surgical technique for the robotically enhanced, minimally invasive treatment of coronary artery disease promote an optimistic way of thinking about the further development of these procedures. PMID- 11005592 TI - Clinical use of a computer-enhanced surgical robotic system for endoscopic coronary artery bypass grafting on the beating heart. AB - The aim of the study was to perform endoscopic coronary artery bypass grafting on the beating heart using a surgical robotic system. In the study, the surgical system ZEUS was used in combination with 3D visualization for endoscopic coronary artery bypass grafting in 25 patients. In a total of 10 cases, the coronary artery anastomosis was done on the beating heart using endoscopic stabilizers without cardiopulmonary bypass. In all cases, total OR time ranged from 4.0 to 8.0 hours (median 5.5 h); the times for endoscopic coronary artery anastomoses ranged from 14 to 50 minutes (median 25 minutes) with no difference between arrested-heart or beating-heart procedures. All patients had an uneventful angiographic control result. An endoscopic coronary artery anastomosis is possible on the arrested as well as on the beating heart. PMID- 11005593 TI - Morphological aspects of Ebstein's anomaly in adults. AB - The essence of the Ebstein's malformation is that the tricuspid valve leaflets do not attach normally to the valve annulus, and the effective orifice is displaced downward into the right ventricular cavity at the junction of the inlet and trabecular components of the right ventricle. Only the septal and posterior leaflets are displaced and divide the right ventricle into two portions. The inlet portion is usually integrated functionally with the right atrium ("atrialized portion"), while the other, including the trabecular and outlet portions, constitutes the functional right ventricle. The proximal atrialized right ventricle often has a wall thinner than the distal functional right ventricle, due to partial congenital absence of myocardium. An atrial septal defect is present in more than one-third of hearts, and the majority of the remainder has a patent foramen ovale resulting in a right-to-left shunt. The downward displacement of the septal tricuspid valve leaflet is associated with discontinuity of the central fibrous body and septal atrioventricular ring, thus creating a potential substrate for accessory atrioventricular connections and ventricular pre-excitation making the patient at risk of sudden death. Angiography has demonstrated that a significant number of patients with Ebstein's anomaly also have morphofunctional abnormalities of the left ventricle, which may be explained by increased fibrosis in the left ventricular wall and ventricular septum as demonstrated by histological studies. Regarding embryology, the leaflets and tensile apparatus of the tricuspid valve are believed to be formed mostly by a process of delamination of the inner layers of the inlet zone of the right ventricle. The downward displacement of the leaflets in Ebstein's anomaly suggests that delamination from the inlet portion failed to occur. PMID- 11005594 TI - Ebstein's anomaly in adults: Doppler-echocardiographic evaluation. AB - A systematic approach and segmental analysis are required for comprehensive assessment including both morphologic and functional abnormalities associated with Ebstein's anomaly. The essence of the disease is an apical displacement of both the septal and the posterior tricuspid leaflets, exceeding 20 mm or 8 mm/m2 in adults. As a consequence, the right heart consists of three components including the true right atrium, the functional right ventricle (RV) and an intervening zone that is anatomically ventricular but functionally right atrial (atrialized RV). The thin wall of the atrialized RV may result in an aneurysm between the anatomic tricuspid annulus and the apically displaced posterior leaflet. The annular attachment of the anterior leaflet is normal, which may be dysplastic and adherent to the RV wall. Tricuspid regurgitation is usually moderate to severe. Size, shape and function of the functional RV must be described. The paradoxical motion of the interventricular septum causes alterations in left ventricular geometry and function. An interatrial communication is frequently present. Echocardiography is the method of choice to diagnose Ebstein's anomaly on its own or in association with other heart defects. PMID- 11005595 TI - Ebstein's anomaly in adults. Arrhythmias: diagnosis and therapeutic approach. AB - Patients with Ebstein's anomaly (M.E.) are known to have a high potential for developing arrhythmia, in the vast majority, of the tachycardia type. Most of these tachycardias are based on accessory pathways (AP) located along the anomalous atrioventricular valve, found in up to 30% of this patient cohort. Next to this main representative for congenital arrhythmogenic substrates, various types of acquired tachycardia were found in patients with M.E., such as atrial ectopic tachycardia (AET), atrial flutter (A. Flutt.), atrial reentry tachycardia (ART), atrial fibrillation and ventricular tachyarrhythmia. Refractoriness to medical treatment and a higher potential for side effects in these patients resulted in an increasing referral to interventional electrophysiology recent years. A total of 37 patients with M. E. and tachycardia underwent an attempt for radiofrequency current (RFC) treatment at our institution, 30 of whom were adults aged 18 to 61 yrs (33 +/- 12.2. yrs). Just 4 patients under went surgery for reconstruction of the anomalous valve and closure of an ASD (2 patients), in whom a total of 6 cases of atrial tachycardia were found, 3 common A. Flutt. and 3 atriotomy-scar related ART. Of the 26 unoperated patients, one presented with typical AV-nodal reentry tachycardia and the remaining patients with atrioventricular reentry tachycardia, based on a total of 39 APs. 27/39 Aps had bidirectional conduction properties, causing the WPW syndrome in 16 patients, 8 APs were concealed and the remaining 4 were Mahaim fibers. Within 39 sessions, 26 of the 30 patients were successfully treated by RFC ablation of the arrhythmogenic substrates. Mean session duration lasted for 313 +/- 151 min (range 95 to 660 min), and a fluoroscopy time of 56.0 +/- 38.4 min (2.2 to 146.8 min) was required. There were no severe acute complications. CONCLUSIONS: In patients with M. E. not only a high potential for the development of tachycardia, but also a tendency towards multiple arrhythmogenic substrates in the single patient can be ascertained. RFC ablation can be used safely and effectively for treatment for various types of tachycardias in patients with M. E.. Such therapy carries the potential for a definitive treatment and should be taken early into consideration as it meets the requirements this patient cohort. PMID- 11005596 TI - Ebstein's malformation. surgical treatment and results. AB - Ebstein's malformation can be defined as an anomaly of the tricuspid valve existing in the setting of a right ventricular dysfunction. The technique introduced by Carpentier in 1980 is based on the concept of mobilization of the restrictive anterosuperior leaflet associated with a longitudinal plication of the inlet component of the right ventricle. From January 1980 to December 1999, 142 patients underwent surgery. The mean age was 25 +/- 15 years (1-65). Cyanosis was present in 48% and associated lesions in 64% of the patients. Patients were classified using a functional approach according to the severity of the lesions. Mild displacement of the septal leaflet, along with small size of the atrialized chamber was seen in 5% (referred to as Type A). Massive displacement of the septal leaflet, but with normal motion of the anterosuperior leaflet and an extensive atrialized chamber, was seen in 35% (Type B). In 51%, the mural (inferior) leaflet was absent, the anterosuperior leaflet was severely restricted by muscular trabeculations and very short tendinous cords, and the anterolateral papillary muscle was incorporated in the right ventricular wall. In these patients (Type C), the atrialized chamber was markedly enlarged and had dyskinetic walls. In such cases, the contractility of the distal (functional) right ventricle was also impaired, and some degree of stenosis of the tricuspid valve was present in one-fifth of them. In the most severe cases (8%), the leaflet tissue of the valve was extremely reduced and the right ventricular walls were thin and contracted poorly. This resulted in the so-called tricuspid sack arrangement (Type D). Valve replacement was needed in only 4 cases, with conservative surgery being achieved in 138 patients by means of mobilization of the anterosuperior leaflet and longitudinal plication of the inlet component of the right ventricle. Additional procedures included the use of a prosthetic ring (94 patients) and partial Glenn anastomosis (30 patients). The hospital mortality was 10%, mainly due to acute postoperative right ventricular failure. Actuarial survival was 75% at 10 years. After operation, 94% of the patients were in functional class I or II of the New York Heart Association, and 88% had no or mild tricuspid valve insufficiency as judged by echocardiography. The rate of reoperation was 9% with a mean delay of 3 years. A second repair was performed in 5 patients. Freedom from reoperation was 87% at 10 years. Sinus rhythm was present in 81%, and 8 pacemaker devices were implanted, 5 for surgically induced atrioventricular block, and 3 because of preoperative conduction disturbances. The use of the partial Glenn anastomosis was introduced recently in cases where the right ventricular contractility was severely impaired, and/or tricuspid valve repair was difficult, and/or permanent atrial fibrillation was present. In those patients with high risk, adding partial Glenn anastomosis reduced the operative mortality from 24% to 6%. Another benefit of the cavo-bipulmonary anastomosis was better functional tolerance of mild residual tricuspid valve incompetence. Those patients with the tricuspid sack arrangement had a high rate of reoperation (2/11) and valve replacement (3/11), but suffered no operative deaths. We conclude that tricuspid valvoplasty associated with longitudinal right ventricular plication is superior to valve replacement. The arrangement producing a tricuspid sack is not suitable for conservative surgery. An associated cavo pulmonary anastomosis decreases the operative mortality in patients at high risk, and seems to preserve right ventricular function. PMID- 11005597 TI - Complete transposition of the great arteries: surgical concepts for patients with systemic right ventricular failure following intraatrial repair. AB - One of the most serious late complications of the intraatrial baffle procedure (Mustard or Senning correction) in patients suffering from transposition of the great arteries, (TGA) is the late systemic right ventricular failure. Nearly all patients presenting with right ventricular dysfunction have severe associated tricuspid regurgitation. The surgical options for these patients include tricuspid valve reconstruction or replacement, staged conversion to the arterial switch operation and orthotopic heart transplantation. Review of 189 operative survivors who underwent the Mustard or Senning operation for TGA between 1970 and 1993 in our institution revealed 12 patients (6.3%) who died from severe systemic right ventricular dysfunction (mean follow-up 16+/-3.5 years), which was the most common cause of late death. All of them had concomitant severe tricuspid regurgitation. 7 patients (3.7%) died from sudden cardiac death. The actuarial survival at 10 years is 93% for simple TGA and 85% for TGA associated with ventricular septum defect or coarctation. At our institution, 4 adolescent or adult patients underwent tricuspid valve surgery; tricuspid valve replacement was performed in 2 patients and valve repair in 2 patients. In the mid-term follow up, 2 of these patients died. Two additional patients underwent orthotopic heart transplantation, and one died on the waiting list. Staged conversion from the Senning/Mustard atrial repair to the arterial switch operation was initially reported by Mee. The procedure for pulmonary artery banding starts with inducing left ventricular reconditioning with subsequent arterial switch. The mortality of this two-staged procedure was as high as 20% to 30% in our early experience, and some of the candidates underwent heart transplantation. Tricuspid valve repair or replacement do not improve right ventricular function in patients with a failing right ventricle following the Mustard/Senning operation. Staged conversion to arterial switch may improve right ventricular function by decreasing the work load of the right ventricle and provides anatomic repair with left ventricle-to aorta continuity. Orthotopic heart transplantation is the only alternative if the left ventricle does not respond to pulmonary artery banding. PMID- 11005598 TI - 25 years after the first arterial switch procedure: mid-term results. AB - BACKGROUND: 25 years after the first successful arterial switch operation (ASO) for complete transposition of the great arteries (TGA), it is possible to evaluate the mid-term results after this procedure. PATIENTS AND METHODS: From March 1986 through December 1993, 188 patients with TGA underwent ASO (primary in 181 patients and secondary after previous banding of the pulmonary artery in 7 patients). Of the 181 patients with primary ASO, 152 had simple TGA or TGA with a small ventricular septal defect, 90 patients had TGA with a large ventricular septal defect, and 10 had a Taussig-Bing complex. Unselected subgroups of the patients were specifically examined with respect to general health, cognitive and motor development, exercise tolerance and cardiological status. Follow-up time was over 5 years except for the cardiac catheterizations, which were performed one year after ASO. RESULTS: Early mortality was 6.4% in the total group and 5.5% in the 181 patients with primary ASO. 5 patients (2.7%) died late and 5 patients needed reoperation during the follow-up period. Cumulative survival for the whole group was 91% at 5 and 10 years after ASO. 96% of the patients were unlimited in their physical activity, and 99% received no medication. Intelligence was not different from a normal population, but motor and neurological impairment was more frequent (23% and 9%, respectively). The Bruce treadmill test was normal in 47 of 50 patients, but provoked signs of myocardial ischemia or salvos of ventricular tachycardia in 3. ECG and Holter ECG showed normal sinus rhythm in 94.5%, important ventricular ectopy in 2.8% and ischemic changes in 1.4%. Right heart catheterization in 67 patients revealed flattening of the pulmonary trunk and frequently mild or rarely moderate pulmonary artery branch stenosis. Left heart catheterization in 71 patients demonstrated significant enlargement of the aortic annulus and root as well as frequent mild aortic insufficiency. One child had unexpected coronary artery occlusion,and 2 showed a hypoplastic left anterior descending coronary artery. In all patients except one, left ventricular ejection fraction was normal, but 6 patients had regional wall motion abnormalities. CONCLUSION: These results and those of other groups are encouraging, but prospective studies over a much longer follow-up time are necessary before definite conclusions can be drawn. PMID- 11005599 TI - 40 years after the first atrial switch procedure in patients with transposition of the great arteries: long-term results in Toronto and Zurich. AB - The atrial switch procedure dramatically improved the prognosis of children with complete transposition of the great arteries (TGA). Overall actuarial survival was approximately 75% after 25 years and was better in patients with simple TGA than in those with complex TGA. Mortality by any cause (16%) and cardiovascular mortality (12% and 13%) were comparable in both centers. Progressive congestive heart failure and sudden death were the principal modes of death. Most of the survivors denied any symptoms or had mild limitations in their daily activities. However, long-term problems in this growing population of adults are challenging and include late arrhythmias (up to two thirds of the patients), systemic ventricular (SV) failure, systemic atrioventricular valve regurgitation and reoperations, such as baffle reconstruction, being the most frequent. Objective assessment of SV function obtained by echocardiography is difficult. It may include fractional area change and tricuspid annular motion. Survivors after an atrial switch procedure are unique and have a good quality of life. However, the definitive and true history of the RV supporting the systemic circulation is not as yet known. PMID- 11005600 TI - Primary myxoid sarcoma of the pleura: 5 years follow-up. AB - Primary sarcomas of the pleura are rare intrathoracic tumours. We describe a case of a recurring myxoid pleural sarcoma in a 51-year-old male patient with coronary heart disease, who initially was admitted for coronary revascularization surgery. Tumour diagnosis was based on pathological examination of a biopsy after radiological detection of a nodule in the right upper lobe. The nodule was excised. The first recurrence was diagnosed after 15 months. Pleuropneumonectomy was done. Four years after the first excision further tumour masses appeared above the diaphragm and were surgically removed and adjuvant irradiation carried out. A year later the patient is doing well. PMID- 11005601 TI - Extended left pneumonectomy combined with off-pump coronary revascularization (CABG). AB - Here, we will report on a 63-year-old patient with a centrally located squamous cell carcinoma of the left lung with complete atelectasis and tumor infiltration into mediastinal pleura and fat. A known coronary two-vessel disease, which became highly symptomatic during the operation made a change of the planned surgical strategy necessary. PMID- 11005602 TI - Reinforcement of a bronchial stump in VATS lobectomy. AB - Bronchial stump reinforcement is sometimes required for patients who have a high risk of developing a bronchial fistula. A lobectomy by video-assisted thoracic surgery (VATS) is widely used for the treatment of both stage I primary lung cancer and metastatic central lung cancer, but there has been no report on reinforcement of a bronchial stump in a VATS lobectomy. We report an aged patient with stage I primary lung cancer concomitant to diabetes mellitus who was successfully treated by VATS lobectomy with reinforcement of the bronchial stump using a pericardial fat pedicle flap. A Harmonic scalpel was very useful and effective in harvesting the fat pedicle. PMID- 11005603 TI - Omental flap for mediastinitis after median sternotomy in asplenia syndrome and gut malrotation. AB - We describe a 12-year-old patient with asplenia syndrome and gut malrotation who, after an interim step before a modified Fontan operation, developed life threatening mediastinitis. A flap of the omentum arising from the transverse colon, longitudinally located in the left of the abdomen, was created and transferred to the mediastinum after a division of the arterial arcade at its most caudal extent. Her postoperative course was uneventful, and 12 months later, the modified Fontan operation was successfully completed. Although visceral heterotaxy results in an omental deformation, it does not preclude the use of an omental flap. PMID- 11005604 TI - Bilobectomy and amphotericin B in a case of endobronchial mucormycosis. AB - Lung mucormycosis is a rare fungal infection that has been described mainly in oncologic and diabetic patients. We here report the case of an endobronchial lesion caused by this fungus in an immunocompetent person. Prompt diagnosis, and therapy with Amphotericin B and surgery, permitted the complete resolution. PMID- 11005605 TI - Optimizing cerebral oxygenation during cardiac surgery: a review of experimental and clinical investigations with near infrared spectrophotometry. AB - OBJECTIVES: Near-infrared spectrophotometry (NIRS) is a promising method for non invasive monitoring of cerebral oxygenation and hemodynamics. This paper reviews studies in which we aimed to validate NIRS in an experimental model of cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) (validation study), use the method in experimental settings to optimize cerebral oxygenation during CPB (oxygenation study), and test its utility during routine cardiac surgery (clinical study). METHODS: Validation study: Forty 8-10 kg piglets underwent 60 min of DHCA at 15 degrees C, were extubated on the first postoperative day, and sacrificed on the 4th postoperative day for histologic investigations. During CPB and DHCA, the animals were investigated by NIRS (monitoring of cerebral oxygenated hemoglobin (HbO2) and oxidized cytochrome aa3 (Cytox)) and magnetic resonance spectroscopy (MRS) (monitoring of cerebral adenosine triphosphate (ATP) and phosphocreatine (PCr). Oxygenation study: A normoxic (n = 5) and a hyperoxic group (n = 5) of piglets underwent 120 min of DHCA and 6 h of reperfusion with NIRS monitoring. Neuronal damage was evaluated by histology. Clinical study: Patients (n = 41) undergoing routine cardiac surgery were investigated by NIRS and neuropsychological testings. RESULTS: Validation study: Reductions of CytOx and HbO2 values were closely correlated with decreases in ATP, PCr, and pHi. The changes in CytOx and PCr showed the strongest correlation (r = 0.623). Maximal CytOx reduction during DHCA predicted brain damage. Oxygenation study: Histology revealed a significant increase in brain damage in the normoxic group. Cytox and HbO2 tended to be lower during DHCA (p = 0.16). Clinical study: During CPB, arterial PCO2, pH and temperature were closely related to CytOx and HbO2. Patients who suffered from neuropsychological deficits had a lower CytOx minimum compared to those without these. CONCLUSIONS: NIRS measurements, especially the CytOx signal, correlate well with high energy phosphates and have a high sensitivity to predict histologic brain damage. NIRS is an important and useful tool in studies investigating cerebral oxygenation during CPB. The CytOx signal predicted impaired neuropsychological outcome in patients. Therefore, the CytOx signal holds much interest for future studies. PMID- 11005606 TI - Cardiac surgery in Germany during 1999. PMID- 11005607 TI - Development of liposome encapsulated clindamycin for treatment of acne vulgaris. AB - The enhancement of topical delivery of hydrophilic substances by use of multilammelar liposomes was measured ex vivo on pig ear skin and in vivo on hairless mice by electron paramagnetic resonance method (EPR). Multilamellar liposomes with different lipid composition (final concentration of membrane components is 48 mg/ml) were loaded with a hydrophilic spin probe GluSL, which does not penetrate the liposome membrane easily. They were characterized with respect to their stability, entrapped volume and enhancement characteristics. We observed significant differences in the properties of different types of liposomes with respect to their stability when in contact with the skin and their penetration into the skin. The results measured in vivo are consistent with those obtained ex vivo. On the basis of these findings the liposomes with appropriate stability and intradermal penetration characteristics were chosen for the development of liposome-encapsulated 1% clindamycin preparation for therapy of acne vulgaris. A double-blind clinical study was conducted to assess the safety and efficiency of liposome-encapsulated 1% clindamycin solution versus 1% clindamycin solution (Klimicin T, Lek). On the basis of the clinical trial it may be concluded that liposome-encapsulated 1% clindamycin solution was therapeutically superior over conventional 1% clindamycin solution in the treatment of acne vulgaris. PMID- 11005608 TI - Vitamin C induced apoptosis in human articular chondrocytes. AB - Chondrocytes present in articular cartilage survive as a resident cell population throughout the lifespan of the individual organism. However, articular chondrocytes as other cells also undergo apoptosis and there is an ever increasing list of diverse stimuli that can induce this phenomenon in vitro. Our main interest was to investigate potential cytotoxic effects of vitamin C (L ascorbic acid) on human articular chondrocytes. The present study suggests that vitamin C can induce apoptosis in a cell culture of chondrocytes after 18 h of cultivation. Apoptosis-inducing activity of L-ascorbic acid is dose dependent and significantly affected by the presence of serum. The increased number of vitamin C induced apoptotic cells was associated with DNA fragmentation and morphological changes of the cells. PMID- 11005609 TI - In vitro functional tests for evaluation of stimulating capacity of cultured human dendritic cells. AB - Basic functional test for evaluation of in vitro cultured human dendritic cells (DC) is primary allogeneic one-way mixed lymphocyte reaction (MLR). In this way, one can evaluate stimulating capacity, which is a basic characteristic of DC. The proliferation of cells is measured through incorporation of 3H-thymidine. Normally proliferation is measured at days 5-7. We studied kinetics of proliferative responses initiated with different stimulating cell suspensions to evaluate differences and possibly reduce time needed to perform this test. Gradual increase in response from days 1 to 7 and a significant difference from controls (peripheral blood mononuclear cells) seen from day 4 was noted if macrophages were used as stimulators. A consistently higher proliferation, compared to controls, was always found already on day 2 when mature DC were used as stimulators. The reaction peaked 2 to 3 days earlier and was also more than two times more intense. This maximal and significantly higher response, consistently seen already after 48 hours, allows us to confirm the presence of mature DC in stimulating suspensions much earlier than previously. PMID- 11005610 TI - Asymmetrical labeling of giant phospholipid vesicles. AB - The method for labeling of inner membrane leaflet in unilamellar giant POPC vesicles was developed and characterised. Symmetrically NBD-PC labeled vesicles were treated by sodium dithionite, which undergoes an irreversible chemical reaction with NBD-PC molecule making it non-fluorescent. After the addition of dithionite the fluorescence on single vesicles as well as on vesicle suspension showed a 50% decrease of its initial value corresponding to marker quenching in the outer leaflet. Hence, fluorimetry as well as fluorescence microscopy prove that dithionite quenching is a suitable method to induce an asymmetrical labeling of the NBD-PC marked giant vesicles. PMID- 11005611 TI - The Str mouse as a model for incontinentia pigmenti. AB - Striated mouse has been proposed as a model for incontinentia pigmenti (IP) based on the similarities in genetic predisposition and syntenic gene localisation in mouse. IP is considered an ectodermal dysplasia with all four characteristic structures involved: sweat glands, hair, teeth and nails. Recently mutations have been found in the Nsdhl, encoding an NAD(P)H steroid dehydrogenase-like protein in Str and Bpa mice. We analysed the phenotype of the Str mouse to evaluate the involvement of ectodermally derived tissues. Our results demonstrated that in Str mouse in addition to abnormal coat texture, sweat glands were severely dystrophic or missing. Retinal degeneration and skeletal abnormalities were also found. We conclude that Str mouse is a good model to get new insights in the pathogenesis of ectodermal dysplasias and X linked male lethality in humans. PMID- 11005612 TI - Animal model in the study of colorectal carcinogenesis. AB - Experimental animal models of neoplastic diseases are important in understanding etiological and pathophysiological processes also in humans. In order to investigate whether the mechanism of genomic instability is associated with chemically induced colorectal tumorigenesis in rat we performed the following study: One hundred and fifty Wistar rats (males 220-280 g and females 140-180 g) were used in the study. Colorectal tumors were induced by means of 15 s.c. applications (20 mg/kg) of 1,2-dimethylhydrazine (DMH). On autopsy, all intestinal lesions were assessed by histological criteria used in human pathology. Forty five tumors were found in the large intestine--30 of these in males and 15 in females, i.e. in 27% of all animals. In four animals multiple primary tumors were found. Histologically 24 tumours were adenocarcinomas, 14 signet-cell carcinoma and 7 adenomas. DNA was extracted from rat neoplastic lesions and adjoining microscopically normal tissues from the same slide and amplified by PCR, using 10 different microsatellite markers from chromosomes 1, 3, 5, 7 and 8. PCR amplicon were analyzed for microsatellite instability with non isotopic method. In 13 adenocarcinomas (29%) microsatellite instability was found at a minimum of 1 locus. Seven tumors (15.5%) showed microsatellite instability at multiple loci. The results of our experiment suggest that genomic instability is an important molecular event in the pathophysiology of DMH induced colorectal carcinogenesis in rats. PMID- 11005614 TI - Soluble tumor necrosis factor receptor I (sTNFRI) as a prognostic factor in melanoma patients in Slovene population. AB - Tumor necrosis factor alpha (TNF-alpha) and its receptors (TNFRI and TNFRII) which exist in soluble form as a product of cleavage of the extracellular domain of membrane integrated receptors, still rise debate about their importance. It was reported that TNF-alpha has numerous actions in diseases such as inflammation, autoimmunity, infectious diseases, septic shock and many types of cancer [1, 2]. Several authors have reported the significance of sTNFRI level in serum of cancer patients [3, 4]. This study was performed in collaboration with the Institute of Oncology of Slovenia. At least two different mouse monoclonal antibodies (MAbs) against human sTNFRI have been prepared to obtain a sensitive and reliable sandwich ELISA. It was compared with commercially available R&D and Endogen ELISAs for the determination of sTNFRI. Groups of patients with different stages of melanoma and epithelial ovarian carcinoma were tested and their clinical records were reexamined. Levels of sTNFRI were measured and compared with the normal serum levels of sTNFRI. PMID- 11005613 TI - Human peripheral blood lymphocytes sensitised to PPD respond to in vitro stimulation with increased expression of CD69 and CD134 activation antigens and production of Th-1 type cytokines. AB - Individuals sensitised to Mycobacterium tuberculosis antigens by infection, vaccination or Mantoux test generate specific memory cells. The response to in vitro restimulation with PPD is observed as the lymphoid cell proliferation and production of Th-1 type cytokines. Cell-mediated immune response was measured by Mantoux test, lymphocyte blast transformation test, estimation of IFN-gamma production (Quantiferon, ELISPOT), and expression of CD69 and CD134 molecules on the T-helper lymphocytes (CD4+). All the methods used were compared for parity of the results. According to Mantoux test results, the patients could be distributed into two groups: responder and non-responder group. Induration in Mantoux test after a new contact with PPD in non-responders was smaller than 5 mm, they produced only small amounts of IFN-gamma, lymphocyte blast transformation was poor, and expression of CD69 and CD134 was low. In responders reaction was much more intensive in all tests measured. We conclude that the reactivity of memory cells to M. tuberculosis antigens can be effectively detected by Mantoux test. The same was true also for the in vitro tests presented but in addition the in vitro tests give more information about the mechanism involved in the immune response against M. tuberculosis. PMID- 11005615 TI - Modulation of tumour necrosis factor production with desmuramyldipeptide analogues. AB - Some synthetic analogues of the immunomodulatory agent muramyl dipeptide (MDP), i.e. phthalimido- (LK-511, LK-413, LK-512, LK-423, LK-508), adamantyl- (LK-415, LK-517), 7-oxoalkyl-(LK-409) desmuramylpeptides were assessed for the tumour necrosis factor (TNF) inducing activity and the ability to modulate TNF production in in vitro phorbol 12-myristate 13-acetate (PMA) & ionomycin stimulated cultures of human peripheral blood mononuclear cells. A kinetic study over a 40-hour period indicated that desmuramyldipeptides were weak TNF inducers compared to romurtide, PMA & ionomycin or lipopolysaccharide. By contrast, they showed the potential to up- or down-regulate the production of TNF evoked by PMA & ionomycin, which was strongly dependent on the time of the stimulation. After 4h of stimulation, the TNF secretion was augmented by LK-508, LK-409 and LK-511, after 18 h by LK-409 and LK-423, and after 40 h by LK-423, LK-511, LK-415 and LK 512. However, LK-517 and LK-512 inhibited the secretion of TNF after the 18-h period. PMID- 11005616 TI - Immune response in lymphocyte cultures stimulated by oral bacteria preparations. AB - Lymphocyte cultures were used as an in vitro experimental model to get a deeper insight into immune response to oral bacteria in periapical granulomas. Lymphocytes isolated from leucocyte concentrate were in lymphocyte cultures stimulated by antigen preparations of oral bacteria. Lymphocyte subsets that have developed in lymphocyte cultures after a week of stimulation were analysed by flow cytometry. A significant increase in expression of INF-gamma molecules in CD3+ cells stimulated by antigen preparations of oral streptococci was found, compared with negative control. On the other hand we observed a significant increase in expression of IL-4 in CD3+ cells stimulated by antigens of anaerobic bacteria, compared with negative control. Our results show that antigens of oral streptococci in in vitro lymphocyte cultures induce the differentiation of T helper cells into Th2 cells and that antigen preparations of anaerobic bacteria induce the differentiation of T helper cells into Th1 cells. Furthermore, an increased expression of HLA-DR molecules on CD8+ T cells stimulated by antigens of oral streptococci was found, compared with negative control. PMID- 11005617 TI - Shift from Th2 to Th1 response in immunotherapy with venoms. AB - Cytokines released from T lymphocytes regulate antibody production by B lymphocytes and releasability of mast cells and basophils. Immune tolerance in specific allergen (Ag) immunotherapy (SIT) might be a consequence of decreased Th2 or increased Th1 response of Ag specific T lymphocytes. We compared function of T lymphocytes in 11 patients with Hymenoptera venom allergy and in 7 healthy volunteers. We followed function of T lymphocytes in 6 patients during SIT. We measured released IL-2, IFN-gamma, IL-4 and IL-5 from Ag and mitogens (either with phorbol-myristate-acetate (PMA) and ionomycin or anti-CD3 antibodies) stimulated peripheral blood mononuclear cells (PBMC). After stimulation with Ag PBMC of patients released 86 +/- 133 pg/ml IFN-gamma and 19.7 +/- 20.1 pg/ml IL-4 (Th2 response), PBMCs of healthy controls released 184 +/- 116 pg/ml IFN-gamma and 4.8 +/- 8 pg/ml IL-4 (Th1 response). Spontaneous release of IFN-gamma in cultures of PBMC of patients increased after rush SIT (before: 25 +/- 31 pg/ml, after 241 +/- 281 pg/ml, p<0.05). After 6 months of SIT response of PBMCs of patients became similar to response of PBMCs of healthy controls. Time pattern of cytokines secreted from PBMCs after stimulation with PMA and ionomycin was different than after stimulation through T-cell receptor/CD3 complex. PMID- 11005618 TI - Interleukin-8 and procalcitonin in early diagnosis of early severe bacterial infection in critically ill neonates. AB - We studied the value of serum interleukin-8 (IL-8) and procalcitonin (PCT) in the early diagnosis of early severe bacterial infection in 58 critically ill ventilated neonates. ELISA was used for determining IL-8 and immunoluminometric assay for PCT. IL-8 and PCT were compared with routinely used serum C-reactive protein (CRP). Neonates were divided into four groups: Ia--proven severe bacterial infection (n = 9), Ib--clinical sepsis (n = 16), II--respiratory distress without bacterial infection (n = 12), and III--various types of neonatal distress (n = 21). Sera were collected on admission, at 24 h and 48 h after admission. There was no significant difference between groups Ia and Ib for either parameter at any time interval. Significant difference was found between group Ia+b (septic neonates) and group II for PCT and CRP at 24 and 48 h, but not for IL-8. There was no difference between group Ia+b and group III except for CRP at 24 h. Diagnostic accuracy was best for PCT on admission and for CRP at 24 h. Serum PCT and IL-8 are not specific markers for early severe bacterial infection in critically ill neonates and are not better than CRP. PMID- 11005619 TI - Interleukin-6 (IL-6) and soluble receptors for interleukin-2 (sIL-2R) in the diagnosis of early severe infection in the critically ill newborns. AB - The aim of our study was to evaluate the diagnostic accuracy of serial determination of interleukin-6 (IL-6) and soluble receptors of interleukin-2 (sIL 2R) in the diagnosis of early infection in the critically ill newborns and compare it with the routinely used C-reactive protein (CRP). Fourty-six critically ill newborns (median age 8 h, range 1-96 h), treated at the multidisciplinary intensive care unit, Division for Paediatric Surgery and Intensive Care, University Medical Centre Ljubljana, were included in the study. Newborns were divided into three groups: group I microbiologically confirmed severe infection (n = 14), group II suspected but not confirmed infection (n = 12) and group III respiratory distress syndrome without laboratory signs of infection. Serum concentrations of IL-6, sIL-2R and CRP were determined on admission and at 12 and 24 h after admission. On admission the concentrations of IL-6 and sIL-2R were significantly higher in group I than in group III, but there was no difference between groups I and II. On admission area under receiver operating characteristic (ROC) curve for IL-6 was 0.756, for IL-2R 0.821 and for CRP 0.799. Repeated determination at 12 h improved diagnostic accuracy for sIL-R and CRP but not for IL-6. PMID- 11005620 TI - Murine monoclonal antibodies directed against human recombinant Macrophage Migration Inhibitory Factor. AB - Macrophage Migration Inhibitory Factor (MIF) is a crucial component of the immune system acting together with glucocorticosteroids to regulate immunity and inflammation. Understanding of its many putative functions and action mechanisms is still ambiguous. Due to the newest findings that a local MIF expression is up regulated in allograft rejection and in glomerulonephritis, an interest in MIF research is increasing and is focused on possibilities of anti-MIF treatment. In the present work new murine monoclonal antibodies (MAbs) directed against human recombinant MIF (hrMIF) are described. hrMIF protein used for the immunisation was tested for its biological activity and has evident macrophage migration inhibitory activity. The selected MAbs were purified and further characterised. They recognised MIF in a Western blot experiment after a native IEF. Anti-MIF MAb designated as M1 inhibited MIF activity in the test, which was performed in the 48 well Boyden chamber system. It is presumed that M1 MAb could be used as a potential therapeutic agent. PMID- 11005621 TI - Effects of molgramostim, filgrastim and lenograstim in the treatment of myelokathexis. AB - Myelokathexis is a very rare form of chronic hereditary neutropenia resulting from impaired neutrophil releasing mechanism in the bone marrow. The recombinant human granulocyte-macrophage (molgramostim) and granulocyte (filgrastim, lenograstim) colony stimulating factors release the mature granulocytes from the bone marrow. We describe a 43-year-old woman suffering from myelokathexis, with the absolute neutrophil count ranging between 0.03 and 1.35 x 10(9)/L. In the period before the introduction of cytokines, the patient had more than 80 major infectious episodes. Since 1991, infections in this patient have been treated with cytokines, given in conjunction with antibiotics. Initially, she received molgramostim in a daily dose of 5 microg/kg subcutaneously, which stimulated the release of granulocytes from her bone marrow, thereby allowing successful treatment of infection. After the development of hypersensitivity, molgramostim was substituted with filgrastim. Finally, lenograstim was given a trial. With all three cytokines, the patient's neutrophil count always attained normal values already 4 hours after subcutaneous application of the drug in a dose of 5 microg/kg, the highest neutrophil levels were measured at 24 hours post injection, and the neutrophil count was again close to the baseline value 72 hours after the treatment. A slight neutropenia was present 48 hours after the application of filgrastim. We believe that all three cytokines are equally effective in increasing the neutrophil count in venous blood of patients with myelokathexis. PMID- 11005622 TI - Expression of human lymphotoxin alpha in Aspergillus niger. AB - A gene-fusion expression strategy was applied for heterologous expression of human lymphotoxin alpha (LTalpha) in the Aspergillus niger AB1.13 protease deficient strain. The LTalpha gene was fused with the A. niger glucoamylase GII form as a carrier-gene, behind its transcription control and secretion signals. Special attention was paid to the influence of different codon usage on secretion of protein. In the case of human tumor necrosis factor alpha (TNFalpha) a dramatic change of secretion has been observed when human cDNA sequence was used instead of synthetic E. coli biased codons. In the case of LTalpha such a change of codon usage brought improvement at the RNA level, however, no increase in the quantity of secreted protein was observed, due to the proteolitic activity of the host organism. The estimated yield of secretion of LTalpha from A. niger into the soya medium was 50 pg l(-1) of culture. PMID- 11005623 TI - Immune response in chronic periapical parodonititis. AB - The purpose of our study was to investigate the immune response in chronical periapical parodontitis (CPP) by using multidisciplinary approach. 30 CPP samples were obtained after surgical removal--apicoectomy. Each CPP sample was examined by histological, bacteriological and flow cytometrical (FC) analysis of lymphocytes infiltrating CPP samples. Ten percent of bacteriological samples were sterile, others had significant aerobic and anaerobic growth. We used pathohistologic and microbiologic findings and compared them to the results of immunological analysis. By FC we found a significant increase in proportions of T lymphocytes expressing interleukin-2 receptors and ICAM-1 compared to peripheral blood lymphocytes. Proportions of T helper cells that produce interferon-gama (IFN-gamma) was higher in CPP samples predominantly colonized by anaerobic bacteria. There were no differences in IL-4 expression by T cells in both groups (anaerobic and streptococcal). Among anaerobic CPP samples differences in proportion of T cells that express IL-2 receptors expression was also found between samples colonised by P. acnes and Bacteroides sp. Oral streptococci cause relatively limited tissue destruction and induce Th2 type of immune response accompanied by non-cytotoxic inflammatory reaction. On the contrary, anaerobic bacteria induce Th1 type of immune response that cause more severe inflammatory reaction (type 4) of hypersensitivity that damage the tissue by the action of cytotoxic T cell activation. PMID- 11005624 TI - Diminished Th1-type cytokine production in gastric mucosa T-lymphocytes after H. pylori eradication in duodenal ulcer patients. AB - Helicobacter pylori infects an estimated 50% of the world population, however only a small proportion of individuals develop clinical symptoms of gastritis, peptic ulceration or gastric cancer. The variations in disease presentation may be due to differences in bacterial virulence and/or immune response to the pathogen. In the previous study we reported an increased expression of the IL-2 receptor in duodenal ulcer (DU) patients infected with H. pylori. This study examined intracellular lymphokine production in gastric mucosa infiltrating T lymphocytes in DU patients before and after H. pylori eradication. T lymphocytes were isolated from gastric mucosa biopsies by using mechanical and enzymatic tissue desegregation. Ficoll-purified lymphocytes were incubated with monoclonal antibodies and analysed by using 3-colour flow cytometry analysis for intracellular interferon gamma (IFNgamma) and interleukin 4 (IL-4) expression in order to define Th1 and Th2 cell population. We demonstrated a significant decrease in the proportion of Th1 cells infiltrating gastric mucosa 6 and 12 months after H. pylori eradication. Our results suggest the importance of the local immune response in the development of H. pylori related gastritis. PMID- 11005625 TI - A mouse model of chronic bacterial lesions (a cotton trap) for studying oral bacteria-lymphocyte interactions. AB - We established a mouse model of chronic bacterial infection (cotton trap) to get a deeper insight into interactions between immune cells and bacterial strains, that are most commonly isolated from periapical processes. We have used flow cytometry to identify the presence of intracellular cytokines of activated T cells collected from cotton traps, previously infected with different strains of bacteria and implanted subcutaneously into the back of the mice. We provide an evidence that anaerobic bacteria (Bacteroides sp.) and nocardiae are more effective in inducing cytotoxic immunity and Th1 response compared to oral streptococci. Differences in immune response against anaerobic bacteria when compared to streptococci are probably dependent on some non-specific immune cell stimulation (e.g. by bacterial cell wall components), nevertheless the role of specific antigen-dependent immune mechanism can not be excluded. PMID- 11005626 TI - Changes in Th1/Th2 cytokine balance in Graves' disease. AB - Graves' disease (GD) is characterised by hyperthyroidism, caused by stimulatory thyrotropin receptor (TSHR) antibodies. Recent research shows that an important factor in the pathogenesis of autoimmune diseases is the change in the balance between Th1 cytokines, which promote cell mediated immunity, and Th2 cytokines, which promote humoral immunity. There are contradictory data about this balance shift in GD. Our objective was to determine the Th1/Th2 cytokine balance shift in patients with newly diagnosed GD, when compared to the same balance in healthy controls. We isolated mononuclear cells (MNC) from the peripheral blood of healthy donors and from patients with newly diagnosed GD before treatment. The MNC were activated with ionomycin in combination with phorbol 12-myristate 13 acetate (PMA). After 40-hour incubation, the concentrations of the cytokines produced (IFN-gamma, IL-4, IL-10, IL-12) in the culture supernatants were measured by ELISA (Endogen, USA). The MNC cultures from patients with GD produced significantly less IL-12 and significantly more IL-10 and IL-4 than MNC cultures from healthy controls. All calculated ratios of Th1 against Th2 cytokines in MNC cultures from patients with GD were significantly lower than in MNC cultures from healthy controls. Our results show a systemic shift of cytokine production in patients with GD toward the Th2 cytokine response, thus confirming the key role of TSHR antibodies and humoral immunity in the pathogenesis of GD. PMID- 11005627 TI - Cytokine-regulated secretion of nerve growth factor from cultured rat neonatal astrocytes. AB - Interactions involved in the regulation of nerve growth factor (NGF) release by inflammatory cytokines: interleukin-1beta (IL-1beta), interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) were examined in rat neonatal cortical astrocytes in primary culture. Exposure of cultured astrocytes to IL-1beta, IL-6, and TGF-beta1 resulted in the stimulation of NGF secretion. Treatment of cells for 24 h with IL-1beta (10 U/ml), IL-6 (5 ng/ml) and TGF-beta1 (5 ng/ml) caused 3-, 1.8-, and 2.8-fold increase in NGF secretion as compared to the control cells. In contrast, TNF alpha (30 ng/ml) by itself had no stimulatory action on NGF release whereas co stimulation of astrocytes with IL-1beta and TNF-alpha showed a synergistic interaction. Co-treatment of astrocytes with IL-1beta and TGF-beta1 increased NGF secretion in an additive way, whereas simultaneous application of IL-1beta and IL 6 resulted in the inhibitory effect on NGF secretion. Our results suggest that interactions between cytokines used cause the stimulation of NGF secretion through different regulatory mechanisms. PMID- 11005628 TI - The effects of histamine and interleukin-6 on NGF release from cortical astrocytes in primary culture. AB - The influence of neurotransmitter histamine and cytokine interleukin-6 (IL-6) on nerve growth factor (NGF) release was studied in rat neonatal cortical astrocytes in primary culture. Exposure of astrocytes to either histamine or IL-6 resulted in the stimulation of NGF release. Maximal stimulation of NGF release was obtained using histamine at concentration 100 nM after 24 h of treatment (2.3 fold increase over the basal secretion from the control cells). IL-6 (30 ng/ml) induced NGF secretion was 1.66 fold over the basal level. Time course of NGF release, after histamine or IL-6 treatment, showed elevation of NGF level in the culture medium after 8 h or 24 h, respectively. IL-6 antibody effectively blocked the IL-6 stimulatory effect on NGF release, but did not influence NGF release, evoked by histamine. IL-6 antibody alone did not show any influence on NGF release. Our results suggest that IL-6 and histamine stimulate release of NGF by two different and independent molecular pathways. PMID- 11005629 TI - Effects of ammodytin L on miniature and endplate potentials in neuromuscular junction of frog m. cutaneus pectoris. AB - Neurotoxic effects of ammodytin L (AtnL), a potent phospholipase A2 homologue, was studied in frog neuromuscular preparation m. cutaneous pectoris by measuring the influence of the toxin on the amplitude and the frequency of miniature and endplate potentials (MEPPs, EPPs). AtnL, in 100 nM concentration, significantly increases spontaneous quantal acetylcholine release from the motor nerve endings, observed as the increase in MEPPs frequency. At 100 nM or higher concentration the toxin decreases EPPs amplitude and the membrane potential (MP) simultaneously. No significant effect of AtnL on EPPs was observed at any concentration bellow 100 nM. Our results indicate that in frog AtnL shows the typical myotoxic effects, but it also exerts presynaptic effects. PMID- 11005630 TI - Subchronic liver injuries caused by microcystins. AB - The subchronic effects of cyanobacterial lyophilizate (CL) containing microcystins on liver were investigated in female New Zealand rabbits. Sterilised CL containing microcystins was injected i.p. Liver toxicity was assessed by histological examination of liver samples. Non-invasive magnetic resonance imaging (MRI) of liver was also performed in order to assess changes in the homogeneity of liver tissue. Subchronical intoxication with microcystins caused morphological changes of liver tissue that were also detected by use of MRI. Histological analysis showed that changes seen on MRI represent liver injury characterised with fatty infiltration and periportal fibrosis. This demonstrates that subchronic exposure to microcystins can lead to liver degeneration, which can easily be detected in vivo by use of MRI. PMID- 11005631 TI - The influence of quantal content on the time course of the endplate current in frogs. AB - The aim of this study was to find out whether control and facilitated endplate current (EPC) of a curarized frog nerve-muscle preparation stimulated with paired impulses decay at the same time constant. The experiments revealed that the decay of the greater second EPC was slower in nearly half of the cases. This half of the results suggests that a larger quantity of the released transmitter and, consequently, a higher density of open channels promote a cooperative effect among them. The end effect can be a longer mean lifetime of open acetylcholine channels. In the other half of the experiments, the mean lifetime of open channels seems to be independent of the agonist concentration. PMID- 11005632 TI - Sensitivity of peripheral nerve fibres to sodium pentobarbital anaesthesia in rat. AB - The sensitivity of different classes of peripheral nerve fibres to anaesthesia with sodium pentobarbital in rat was tested. The basic approach was to examine changes in compound action potential (CAP) in a rat sural nerve induced by sodium pentobarbital. Rats were either sacrificed by cervical dislocation (control) or anaesthetized with sodium pentobarbital (100 mg/kg, i.p.), and a 30 mm long sural nerve segment excised and placed on electrodes in a thermostatically controlled recording chamber. CAPs were evoked by electrically stimulating the sural nerve segment with supra-threshold stimuli. CAP in control sural nerves consisted of the early component (A axons: 32.73 +/- 2.91 m/s) and the late component (C axons: 0.92 +/- 0.05 m/s) with peak voltage amplitudes 4.9 +/- 1.0 mV and 0.1 +/- 0.03 mV, respectively. Anaesthesia with sodium pentobarbital had no effect on CAP latency. However, the amplitude of CAP of both A and C axons was reduced by approximately 40% and 50%, respectively. The depressant effect of pentobarbital on CAPs was statistically significant for both groups of axons (p<0.01). Non selective sensitivity of A and C axons to pentobarbital suggests even distribution of receptors for GABA in these two populations of axons in the rat peripheral nerve. PMID- 11005633 TI - Phosphate metabolism in red blood cells of critically ill neonates. AB - Very few data exist on phosphate metabolism in critically ill neonates. Therefore we studied the incidence of hypophosphataemia, the intracellular metabolism of phosphate by measuring adenosine 5'-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) in red blood cells, and excretion of phosphate in urine. The aims of the study were early detection of changes in phosphate metabolism as possible diagnostic markers of sepsis and defining the cause of hypophosphataemia. Neonates, treated in multidisciplinary paediatric intensive care unit (PICU), included in the study, were less than three days of age. Eighteen of them had respiratory distress syndrome (RDS) and 16 had microbiologically confirmed or clinical sepsis. The overall incidence of hypophosphataemia in critically ill neonates was over 80%, and was more common (88%) and more profound in those with sepsis than in those with RDS (79%). Therefore the septic neonates needed significantly larger amounts of phosphate to maintain normophosphataemia. In septic neonates ATP concentration in red blood cells was significantly lower than in neonates with RDS and controls, while the 2,3-DPG concentration was increased as a result of compensation. In septic neonates urinary losses of inorganic phosphate (Pi) were significantly higher than in neonates with RDS. Hypophosphataemia in critically ill neonates is at least partly due to higher urinary losses of phosphate. PMID- 11005634 TI - Effect of mechanical load on acetylcholinesterase mRNA levels in the slow soleus muscle of the rat. AB - The influence of mechanical load on the levels of acetylcholinesterase (AChE) mRNA was studied in order to examine to which extent different loading conditions may be responsible for differences in AChE regulation between the soleus, which is an antigravity muscle, and the fast EDL muscle. Forty-eight female rats were randomly assigned to three groups: a group with hindlimb suspension producing soleus muscle unloading, a group with ablation of synergistic gastrocnemius muscle causing overload of the soleus muscle, and the control group. The soleus muscles were isolated after 8 days of treatment AChE mRNA levels were analyzed by Northern blots and evaluated densitometrically. The values were normalized with beta-actin mRNA level, and then a value of 100% was assigned to the mRNA level in the control EDL muscle. Muscle unloading did not produce a significant increase of the AChE transcript levels, but the levels were rather variable. However, a statistically significant increase of AChE mRNA levels was observed in overloaded soleus muscles. These results corroborate the hypothesis that the slow and fast patterns of activity appear more important then muscle loading for the differences in regulation of AChE mRNA levels in fast and slow muscles. PMID- 11005635 TI - Subunit interface selective toxins as probes of nicotinic acetylcholine receptor structure. AB - The pentametric assembly of the nicotinic acetylcholine receptor with two of the five subunit interfaces serving as a ligand binding sites offers an opportunity to distinguish features on the surfaces of the subunits, and their ligand specificity characteristics. The receptor from mammalian muscle, with its circular order of homologous subunits (alphagamma alphadelta beta), assembles in a unique arrangement. The residues governing assembly can be ascertained through mutagenesis. Selectivity of certain natural toxins is sufficient to distinguish between sites at the alphagamma and alphadelta subunit interfaces. By interchanging residues on the gamma and delta subunits through mutagenesis, and ascertaining how they interact with the alpha subunit, determinants forming the binding sites can be delineated. The alpha-conotoxins show a 10,000-fold preference for the alphadelta over alphagamma subunit interface with alphaepsilon falling in between. The waglerins show a 2,000-fold preference for alphaepsilon over the alphagamma and alphadelta interfaces. Finally, the alpha-neurotoxin from N. mossambica mossambica shows a 10,000-fold preference for the alphagamma and alphadelta interfaces over alphaepsilon. Identification of interactive residues through mutagenesis, when coupled with homology modeling of domains and site directed residue modification, has revealed important elements of receptor structure. PMID- 11005636 TI - Cholinesterase: substrate inhibition and substrate activation. AB - The relationship between activities and substrate concentrations (pS-curves) was analysed for reactions of acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8). Catalytic constants Km, Kss, Vm, n and b were calculated from the Michaelis, Haldane, Hill and Webb equations in order to assess whether a given substrate also acts as an inhibitor or activator. It is suggested that the term substrate inhibition should only be attributed to substrates revealing bell-shaped pS-curves, while the terms apparent substrate inhibition or apparent substrate activation should relate to calculated values of the catalytic constants. PMID- 11005637 TI - Role of alpha-adrenoceptors in the cutaneous postocclusive reactive hyperaemia. AB - Laser-Doppler (LD) flow measurements reveal typical flow oscillations in the descending part of the cutaneous postocclusive reactive hyperaemia (PRH). The origin of these oscillations is still poorly understood. We tested the hypothesis that the high frequency (HF) components within the frequency band 0.06-0.2 Hz are due to the local myogenic mechanism, whereas the low frequency (LF) components within frequency band 0.01-0.05 Hz reflect sympathetic vasomotor activity. LD flow was monitored on fingertips of 11 healthy volunteers before and after an 8 minute occlusion of digital arteries. We studied the effect of intradermal microinjection (1 microl) of alpha-antagonists (prazosine, yohimbine) and alpha agonists (phenilephrine, clonidine) on PRH oscillations. We analysed the magnitude of peak flow and its duration and performed spectral power analysis to obtain fundamental HF and LF frequencies (defined as components with the highest amplitude in the part of the respective frequency band). The results (mean +/- SE) for each substance were compared to the control values obtained after microinjection (1 microl) of 0.9% NaCl. The fundamental HF significantly decreased from 0.11 +/- 0.008 Hz after an injection of saline solution to 0.08 +/ 0.006 Hz after an application of antagonists (p<0.05). The local application of alpha-antagonists did not abolished HF oscillations what suggests that activity of local sympathetic nervous system is not essential in generation of these oscillations. However, the significant decrease of fundamental frequency in HF band after application of alpha-antagonists supports the hypothesis of their local origin and could be explained by the effect on pacemaker myogenic mechanism. PMID- 11005638 TI - Spatial resolution of epicardial pace mapping using body surface potentials. AB - Body surface potential maps (BSPMs) recorded during pace mapping provide an important non-invasive means for identifying local cardiac events; recent clinical studies demonstrated that endocardial pacing sites can be resolved within less than 10 mm. We sought to determine whether similar spatial resolution could be achieved during epicardial pacing. Four patients who were undergoing either heart valve replacement (one), aortocoronary bypass graft (one), or both (two) were studied. In each patient, a pair of epicardial electrodes was placed intraoperatively at the middle aspect of the right ventricular free wall. The distance between the neighbouring electrodes was 10 mm. Five days after the surgery, ECGs were acquired from 35 leads during pacing from each epicardial electrode. We determined the distributions of QRS integrals (the net area under the ECG signal) and compared integrals corresponding to pacing from each of the adjacent electrodes using statistical indices. Student's t-test was applied to these indices and in all the patients revealed that differences in distributions of QRS integral maps were statistically significant (p < 0.01). Results of our study indicate that the non-invasive acquisition of body surface ECGs could resolve epicardial breakthrough sites within 10 mm, which may be useful in facilitating therapeutic ablations in patients with ventricular tachycardias. PMID- 11005639 TI - Improvement of early functional atherosclerotic changes in males with hypercholesterolemia after vitamin E supplementation. AB - Vitamin E as an antioxidant vitamin reduces the susceptibility of low-density lipoprotein (LDL) cholesterol to oxidation and may have antiatherosclerotic effects. We tested the hypothesis that six months of 400 mg vitamin E supplementation favourably affects early functional changes in atherosclerotic process in subjects with hypercholesterolemia. The diameter of the brachial artery at rest, after reactive hyperemia (representing endothelium-dependent vasodilatation) and after sublingual glyceryl-trinitrate (representing endothelium-independent vasodilatation), were determined by ultrasonographic method (B mode) before and after the intervention period. After the intervention period the brachial endothelium-dependent vasodilatation increased significantly in the vitamin E group while it did not change in the placebo group. In conclusion, six months of oral vitamin E supplementation results in improvement of the endothelium-dependent vasodilatation in men with hypercholesterolemia. PMID- 11005640 TI - Protein-polyphenol interactions and in vivo digestibility of buckwheat groat proteins. AB - A rat model system, based on a diet with (Nb+) or without (Nb-) the inclusion of 0.7% of a 2:1 (w/w) mixture of Bacitracin and Neomycin sulfate (Nebacitin) was used to elucidate interactions of polyphenols with proteins in buckwheat. The treatment with the antibiotic Nebacitin suppresses the microflora in the large intestine and is used to obtain an estimate of the digestion occurring in the small intestine. Experimental buckwheat samples were obtained in order to contain different levels of polyphenol and were subjected to different hydrothermal treatments. It was found that a considerable interaction between polyphenols and proteins appeared during the hydrothermal treatment, and this interaction reduces the digestion of proteins through the small and large intestine. Microbial processes in the colon enhance the digestibility of protein, blocked by polyphenols in hydrothermally processed buckwheat. PMID- 11005641 TI - Measurement of porcine heart temperatures. AB - During cardiac surgery local hypothermia of the heart is used to slow down cellular metabolism. It has been shown by computer simulation that heart muscle is not uniformly cooled to the desired temperatures. The aim of our work was to find out a way for measuring the temperatures of the ventricular wall and the septum with +/-0.1 degrees C accuracy. The measurement protocol should be applicable during operations on the human heart, during which disturbing factors should be minimized. We have tested the protocol and the equipment on porcine hearts. Two types of probes were tested, the contact-probe, which can be attached to the heart-wall surface, and the needle-probe, which can be inserted into the heart tissue. Before experiments the probes were calibrated and their heat transfer characteristics determined. We conclude that the probes and the protocol are suitable for studying the effects of cardioplegia, and for checking the results previously obtained by computer simulation. PMID- 11005642 TI - Biochemical and biophysical conditions for blood clot lysis. AB - We have studied how pharmacological dissolution of blood clots was affected by clot retraction, the mode of transport of fibrinolytic agents into the clot and the thickness of the composite fibrin fibers. Retracted clots were resistant to fibrinolysis in a milieu without dissolved plasminogen, because the amount of fibrin-bound plasminogen in retracted clots was insufficient for successful clot lysis. In plasma containing plasminogen, retracted clots were successfully lysed with fibrin-specific plasminogen activators, but not with non-fibrin-specific activators. Preincubation of retracted clots in plasma increased their plasminogen content as well as their sensitivity to fibrinolysis. The rate of lysis was increased up to 100-times when plasminogen activator and plasminogen were introduced into cylindrical clots by pressure-induced bulk flow in comparison with diffusion alone. The magnitude of the increase was similar in retracted and nonretracted clots, but the absolute rate of lysis was faster in non-retracted clots. The influence of fibrin fiber thickness on fibrinolysis was studied by atomic force microscopy. The time to complete lateral section of fibers did not differ between thick and thin composite fibers, and the rate of diameter reduction was faster in thick fibers than in thin ones. Taken together our results suggest that lysis of retracted clots proceeds in circular stages: (a) activation of bound plasminogen followed by partial degradation of fibrin, (b) opening of new plasminogen-binding sites on partly degraded fibrin, (c) binding of plasminogen to the new binding sites which enhances the susceptibility of clots to lysis. Lysis is accelerated by bulk flow of plasminogen activator and plasminogen into clots in comparison to diffusion alone. Fibrinolysis of thick composite fibrin fibers proceeds more efficiently than lysis of thin fibers. PMID- 11005643 TI - Possibilities for clinical use of prostacyclin in vascular disease. AB - Currently available drug therapies for patients suffering severe ischaemia with rest pain and trophic lesions of the limbs remain unsatisfactory. Also vascular reopening procedures are suitable in only about half of the patients. In atherosclerotic disease when the vascular endothelium is damaged prostacyclin synthesis is decreased and thromboxane A2 production increases. Prompted by this knowledge of the importance of prostacyclin in pathogenesis of atherosclerotic disease an attempt was made to employ PGI2 clinically--for treatment of advanced forms of peripheral arterial atherosclerotic disease. Favourable effects of the stable analogue of prostacyclin (Iloprost), were reported in various studies, which included patients with peripheral atherosclerotic arterial disease, thromboangiitis obliterans and Raynaud's phenomenon. The use of Iloprost resulted in a significantly superior response than other drugs and placebo in terms of alleviation of rest pain, ulcer healing and decrease of amputation rate of ischaemic limbs. Therefore prostacyclin provides a therapeutic option in patients with advanced forms of arterial disease--including critical ischaemia. PMID- 11005644 TI - Lacidipine decreases the honeybee venom-induced vasoconstriction of the isolated porcine coronary artery. AB - The venom of the honeybee Apis mellifera induces cardiovascular dysfunction. As its effects on coronary arteries have not yet been described, we studied the effects of the whole honeybee venom (non-volatile part) in the isolated porcine left anterior descending coronary artery (LAD) and the influence of L-type Ca2+ channel blocker, lacidipine, upon the venom effects in LAD. The venom produced concentration dependent contractions (7-70 microg/ml) of the porcine LAD; maximal effect of the venom was approximately the same as the effect of 30 mM KCl. Lacidipine concentration dependently (0.1-10 microM) and significantly (P < or = 0.05) decreased the venom-induced vasoconstriction. The results indicate the involvement of L-type Ca2+ channels in coronary contraction, induced by bee venom. PMID- 11005645 TI - Effects of mibefradil and verapamil on ischemic-reperfusion in the hearts of guinea pigs with acute renal failure. AB - Two parameters indicating the ischemic-reperfusion myocardial injury, coronary flow and lactate dehydrogenase (LDH) release rate, were evaluated in guinea pigs with gentamicine-induced acute renal failure (ARF) and compared with those of healthy animals. Isolated Langendorffs hearts were exposed to 50 min of zero-flow global ischemia and 60 min of reperfusion. The influences of calcium channel antagonists (of T- and L-type antagonist mibefradil and of L-type antagonist verapamil) in reperfusion solution were evaluated. Our results showed coronary dilatation and higher LDH release rate in ARF than in control hearts before ischemia. Recovery of coronary flow during reperfusion was better and LDH release rate lower in ARF vs. control hearts. Perfusion with mibefradil and verapamil did not additionally increase coronary flow, however 0.1 microM mibefradil and verapamil decreased LDH release rate during reperfusion in ARF hearts in comparison to control hearts. Our results showed some protective effects of 0.1 microM mibefradil and verapamil on LDH release rate during reperfusion, but not on coronary flow in guinea pigs hearts with ARF. PMID- 11005646 TI - Thermal conductivity of the porcine heart tissue. AB - The porcine heart was used as a model for studying the thermal changes in myocardium at cooling and re-warming during open heart surgery. A section of the heart septum was excised and tissue was cut into two similar square slices. The same shape of the tissue, cut from the surface from the upper lateral wall of the left ventricle, covered with epicardium and fat, was taken for another measurement. A thin (<0.5 mm) square thermal source of the same length of the side as the tissue samples was put between the two slices of tissue. This set was placed in the middle of two identical copper cylinders (2r=50 mm, height=55 mm) used to keep the outer side of the specimen at controlled room temperature. Thermal conductivity of the heart tissue was determined at controlled thermal power, and known difference of the temperature at the edge of the tissue and at the middle of the heater, when steady state was reached. Thermal conductivity calculated from the temperature difference and the geometry of heater and samples was 0.75 W/m.K for septal heart tissue, and 0.60 W/m.K for the lateral wall ventricle tissue with epicardium and fat. PMID- 11005647 TI - Nicardipine dose-dependently reduces the effect of equinatoxin II on coronary flow in isolated porcine heart. AB - Death after i.v. administration of equinatoxin II (EqT II) has been attributed to the circulatory failure resulting from cardiotoxic effects. The mechanism of action is unknown. The aim of the present work was to study the effects of the toxin on vascular tone in the isolated porcine coronary artery and on coronary flow in the isolated pig heart. EqT II caused concentration-dependent contractions of rings of the isolated epicardial porcine coronary artery with an EC50 value of 89+/-5 nM (n=5-6) and maximal effect of about 140% of the contraction induced by 20 nM KCl. On Langendorffs porcine heart preparation EqT II caused a dose-dependent decrease of coronary flow. At EqT II doses lower than 0.05 micromol/100 g of heart weight there were no measurable effects of the toxin. At dose 0.5 micromol/100 g the toxin decreased coronary flow to less than 9.8+/-2.5% of the control value. The constrictory effect of the toxin on isolated porcine coronary arteries was diminished by the L-type calcium channel antagonist nicardipine (NC). NC in 1 microM concentration almost completely abolished the effect of EqT II on coronary flow. Our results confirmed involvement of L-type calcium channels in the vasoconstrictory effects of EqT II on epicardial coronary arteries. PMID- 11005648 TI - Action of mibefradil and lacidipine on the isolated human anterior tibial artery. AB - In isolated human anterior tibial artery the effects of two different types of calcium channel antagonists, mibefradil (a selective T-type Ca2+ channel antagonist) and lacidipine (a 1,4 dihydropyridine Ca2+ channel antagonist, acting at L- and T-type, but binds preferentially at L-type Ca2+ channels) were compared. Both drugs reduced the contractions of isolated arteries induced by 60 mM KCl. The potency (IC50) of mibefradil was 6.5 microM and of lacidipine 82.4 nM. The potencies of both Ca2+ channel antagonists differed significantly (p<0.001 at 0.1 and 1 microM; p<0.01 at 10 microM). Lacidipine was 79-times more effective than mibefradil in reducing the vasoconstriction in isolated human anterior tibial artery. One of the reasons for higher potency of lacidipine could be a higher density of L-type than of T-type Ca2+ channels in tissue of the human anterior tibial artery. PMID- 11005649 TI - Comparison of effects of nitrendipine, lacidipine and mibefradil on postischaemic myocardial damage in isolated rat hearts. AB - During ischaemia and reperfusion increased cytosolic Ca2+ is one of the important causes for ischaemic-reperfusion myocardial injury. In the present study we compared effects of preferentially L-type Ca2+ antagonists nitrendipine (NT) and lacidipine (LP), and of mibefradil (MB) a Ca2+ antagonist with higher affinity to T- than to L-type channels on myocardial function during reperfusion. Coronary flow (CF), heart rate (HR), left ventricular pressure (LVP), lactate dehydrogenase (LDH) release rate and ECG were registered during 40 min of reperfusion following 30 min of global zero flow ischaemia in Langendorff's isolated rat hearts. Either NT (100 nmol/L) or LP (10 nmol/L) or MB (100 nmol/L) was added to Krebs-Henseleit solution 10 min before ischaemia till the end of experiments. All three drugs influenced CF, HR and LVP. All of them decreased LDH release rate (P < 0.05, in microkat/g x min) when compared with control hearts (53.2 +/- 5.1): MB (19.4 +/- 4.3) > LP (30.7 +/- 6.6) > NT (43.3 +/- 2.8). NT reduced the duration of continuous arrhythmias at the beginning of reperfusion (to 59.1 +/- 6.1% of ischaemic controls) as well as the number of single arrhythmic events arising during the whole period of reperfusion (to 26.1 +/- 6.0% of ischaemic controls). MB diminished only single arrhythmic events during reperfusion to 39.1 +/- 17.3% of ischaemic controls. LP did not affect the onset of arrhythmias. Results of our experiments indicate a relatively greater importance of T-type than of L-type Ca2+ channels in the arising of postischaemic myocardial damage. PMID- 11005650 TI - The importance of hemolysis in the lethal effects of equnatoxin II, a protein from the sea anemone Actinia equina (L.). AB - Equinatoxin II (EqT II) is one of the three isotoxins isolated from the sea anemone Actinia equina (L). The cause of death due to i.v. application of Eq T II was attributed to its hemolytic activity and hiperkaliemia, and the direct cardio respiratory effects of the toxin. The toxin also binds to plasma lipids and forms toxic conglomerates with them. In our study we tried to evaluate the importance of interaction of the toxin with blood components in the toxin lethality. Anaesthetized male Wistar rats were used. The respiratory activity, ECG and animal serum ionic composition were monitored. To simulate isolated effects of the toxin hemolytic activity, hemolysed erythrocytes, KCl and mixture of rat serum with the toxin were injected i.v.. It was shown that neither the lysed erythrocytes nor the mixture of rat serum with the toxin were lethal for the experimental animals. Even KCl, in concentrations detected in serum of rats killed by the toxin, did not cause the death of experimental animals. Our results indicate that either a combination of the tested effects of the toxin on blood components or more probably the direct effects of the toxin on vital organs are responsible for the toxin lethality. PMID- 11005651 TI - Assessment of kainate toxicity using contrast enhanced magnetic resonance imaging. AB - The purpose of this study was to test the capability of contrast enhanced magnetic resonance imaging (MRI) in assessing lesion formation in rat brain after systemic (i.v.) administration of kainate. MRI was performed with T1-weighted spin echo sequence before and after the administration of kainate and contrast media. Contrast media used were based on paramagnetic gadolinium (III) ion: Gd DTPA (gadoliniumdiethylenetriaminepentaacetic acid) and prototype agents for blood-pool enhancement. Gadomer-17 and polylysine-Gd-DTPA. Enhancement of lesion rims and other brain tissue abnormalities due to kainate with Gd-DTPA, Gadomer-17 and polylysine-Gd-DTPA were observed mainly in the region of hippocampus and in the areas not protected by the blood-brain-barrier (BBB). PMID- 11005652 TI - Alpha-melanocyte stimulating hormone desensitizes the responsiveness of carbon fibres within seconds. AB - We monitored electrooxidation of noradrenaline and alpha-melanocyte stimulating hormone (alpha-MSH) at a carbon-fibre microelectrode (CFME). The solution of noradrenaline (1 mM) or alpha-MSH (1 mM) was applied by a pressure pulse (2 s) from a micropipette to a voltage-clamped (850 mV) CFME immersed into bathing solution of an inverted microscope chamber. The distance between the CFME and micropipette was 2 to 12 microm. The maximal currents recorded for these two agents were 8.0 +/- 0.5 pA (N = 9) and 3.0 +/- 1.1 pA (N = 9), respectively. Pressure application of control solution did not affect the measured current. The noradrenaline-evoked anodic current was characterized by a monotonic increase that attained the maximum at the end of the pressure pulse. In contrast, the time course of the alpha-MSH-evoked current was biphasic. The maximum amplitude of this current was attained in 0.59 +/- 0.15 s (N = 9) and then it declined with a time constant of 7.5 +/- 4.0 s (N = 9) until the pressure pulse was terminated. We explain this phenomenon to be due to an interaction between the peptide oxidation products and the CFME which results in its desensitization. PMID- 11005653 TI - Selective recording of neuroelectric activity from the peripheral nerve. AB - Electroneurograms (ENGs) from superficial regions of the sciatic nerve of a dog, innervating the tibialis anterior (TA) and gastrocnemius muscles (GM), arising mainly from muscle spindles and Golgi tendon organs were recorded selectively with an implanted 33-electrode spiral cuff (cuff). Relative positions of superficial regions within the cuff were defined by delivering stimulating pulses on groups of three electrodes (GTEs) within the cuff which were in contact with them. It was found that GTEs eliciting maximum contractions of muscles were GTE No. 3 for the TA muscle and GTE No. 8 for the GM muscle. In the first experiment the implanted leg was mounted into a special electronic brace. Extending forces were applied to the ankle rotating it by up to +/-37 degrees according to the neutral position, thus eliciting torques in the TA muscle of up to 1.2 Nm. Channel 1 of the 4-channel preamplifier was connected to GTE No. 8, channel 2 to GTE No. 2, channel 3 to GTE No. 11 and channel 4 to GTE No. 5. Results show that only ENG recorded with GTE No. 8, being close to the region innervating the TA muscle, correspond to the mechanical load. In the second experiment the calcanean tendon (CT) of an implanted leg was dissected. The proximal end of the CT was connected to a force transducer and repetitive pull forces (about 12 N) were applied to the CT. Channel 1 of the preamplifier was connected to GTE No. 5, channel 2 to GTE No. 1, channel 3 to the GTE No. 11 and channel 4 to GTE No. 8. Results show that only ENG recorded with GTE No. 5, being close to the region innervating the GM muscle, correspond to the mechanical load applied on CT. PMID- 11005654 TI - Seizure-induced changes in energy metabolites and effects of N-tert-butyl-alpha phenylnitrone (PNB) and vitamin E in rats. AB - Impaired energy metabolism may play a critical role in the neuronal injury caused by kainic acid (KA) induced status epilepticus (SE). Following an acute dose of KA (15 mg/kg, s.c.) rats developed SE within 1 h. Rats were sacrificed 1 or 72 h after the onset of SE using a head focused microwave technique and the brain regions (pyriform cortex, amygdala, and hippocampus) were assayed for energy metabolites: ATP, ADP, AMP, phosphocreatine (PCr) and creatine (Cr) using reversed-phase HPLC (RP-HPLC). Control values were significantly higher in cortex (23-32%) than in other brain regions. Within 1 h, SE caused a marked decline in ATP (44-56%), PCr (49-64%), total adenine nucleotides (TAN, 45-50%) and total creatine compounds (TCC, 32-51%). Within three days, the hippocampus showed the greatest recovery, as the reduced values returned to normal. Pretreatment of rats with an antioxidant (PBN, 200 mg/kg, i.p., 30 min prior to KA; or vitamin E (Vit E), 100 mg/kg, i.p./day for 3 days), which did not prevent seizure activity, attenuated depletion of high-energy phosphates caused by KA. These findings suggest that the depletion of energy metabolites caused by KA-induced seizures may be linked to oxidative stress mediated toxicity. PMID- 11005655 TI - Neurotransplantation-induced plasticity in the recipient CNS: focusing on the recipient response. AB - In the last two decades, neurotransplantation has gained the status of a potentially valuable treatment option in various central nervous system (CNS) disorders. This technique has provided considerable functional improvement in animal models of neurodegenerative diseases, stroke and trauma. In order to make the best therapeutic use of this treatment option, mechanisms of neurotransplantation-induced recovery need to be better understood. Specific interactions of transplants with the recipient brain, which are prominent in embryonic neural cell grafts, include formation of synapses between grafted neurons and recipient neuronal population with controlled release of neurotransmitters. Production and release of specific trophic substances for the recipient neurons by the grafted cells also play a considerable role in some transplants. In the above cases, the functional recovery seems to correlate well with the number of surviving cells in the transplant. There is, however, another component to the graft-induced recovery, best revealed in those graft recipients who display functional improvement although only few or no grafted cells can be found at the post mortem analysis. While psychological factors (placebo effect) have been proposed to play a central role in human graft recipients with functional recovery in the absence of surviving grafts, animal models of neurodegenerative disease have consistently shown the same phenomenon. Our recent results point to the local inflammatory and immune response to transplantation as a key element which induces a trophic response in the CNS parenchyma and stimulates plastic changes of the recipient neural connections. Findings by other investigators, who studied the connections between the inflammatory and neurotrophic responses in vitro and in vivo, and glial reaction to CNS trauma and trophic factor synthesis in vivo, support such conclusions. Accumulated evidence point to the need for further studies that would elucidate the role of the immune response in connection with CNS transplantation outcome. PMID- 11005656 TI - Biochemical aspects of Salter's osteotomy for treatment of acetabular dysplasia. AB - The center-edge (CE) angle was measured on serial antero-posterior radiographs of 44 hips treated by Salter's innominate osteotomy for developmental dysplasia of the hip. Radiographic examination according to Severin was performed and the peak hip joint pressure was calculated using geometrical parameters determined from radiographs at the most recent follow-up examination, 7 to 13 years postoperatively. At the latest follow-up assessment, there was a positive correlation between the postoperative CE angle and radiographic results. Negative correlation was found between the postoperative CE angle and peak hip joint pressure at the latest follow up. In conclusion, Salter's osteotomy performed to treat developmental dysplasia of the hip should involve radical acetabular correction resulting in maximum increase of the CE angle. PMID- 11005657 TI - The development of the method for the determination of terbinafine in cat's plasma and hair. AB - Clinical investigations of terbinafine indicate its high treatment activity against infections by several dermatophytes. Its efficiency was tested also in the treatment of microsporosis in cats. The distribution of terbinafine in cat's plasma and hair is important for the identification of the drug efficiency. A fast and reliable reversed-phase high performance liquid chromatographic method with appropriate sample preparation has been developed. Reliability, good reproducibility and low detection limit (LOD 0.25 ng/ml) of the method enable determination of terbinafine in hair and also in plasma of cats infected with Microsporum canis treated by Lamisil tablets. PMID- 11005658 TI - Serum T3 and T4 concentrations in the adult rats treated with herbicide 2,4 dichlorophenoxyacetic acid. AB - The influence of sublethal doses of 2,4-dichlorophenoxyacetic acid (2,4-D) on serum T3 and T4 concentrations in Hsd Cpb: Wistar rats of both sexes was studied. The trial was performed on 24 males and females respectively, each divided into three groups of 8 animals (control, groups 1 and 2). Aqueous solution of the compound (11 mg/kg body weight--group 1 and 110 mg/kg body weight--group 2) or clean tap water (control group) was used. Aliquots of 2.4 ml/kg body weight were administered with a stomach tube from the 1st to 10th day of the experiment. Three days before the first treatment and on the 6th and 13th day of the experiment the serum T3 and T4 concentrations were determined by commercial radioimmunoassay kits (Byk-Sangtec Diagnostica), validated for rats. A significant decrease of serum T4 (P < 0.01) and T3 (P < 0.001) was determined in males of groups 1 and 2 during the experiment. On the 6th day of experiment serum T4 and T3 values were significantly lower (P < 0.001 and 0.01 respectively) in group 2 than in the controls and group 1 of both males and females. During the whole experiment serum T4 levels were lower in females than in males (P < 0.05). PMID- 11005659 TI - In vivo effects of head-to-tail 3-alkylpiridinium polymers isolated from the marine sponge Raniera sarai. AB - Water soluble polymeric 3-alkylpyridinium salts (poly APS; MW 18900 and 5520 Da) were isolated from the marine sponge Raniera sarai. In vitro it strongly inhibited acetyl cholinesterase (AChE) from different species (electric eel, horse serum, human erythrocytes). In our experiments the importance of anti AChE activity in the toxin lethality was evaluated. In vivo experiments were performed on male Wistar rats and ECG, blood pressure and breathing pattern were monitored. After i.v. application of lethal doses of the toxin ECG showed signs of hypo perfusion. Arterial blood pressure fell to mid-circulatory pressure, and breathing stopped after a few breaths At sublethal doses the toxin caused an increase of residual volume, prolongation of expiration, and bradycardia. Patho anatomical examination revealed that the plugs in lung circulation may cause the death of experimental animals due to cardiorespiratory failure. PMID- 11005660 TI - Different operative treatment of slipped capital femoral epiphysis: a comparative study of biomechanical status of the hip. AB - The biomechanical status of the hips subject to two different operative treatments was studied. The biomechanical status was determined by the peak stress on the weight bearing area using the 3-d mathematical model. It was found that the procedure according to Imhauser yields considerably and statistically significantly higher peak stress than the procedure according to Dunn-Fish. PMID- 11005661 TI - Peak contact stress in human hip during gait. AB - The contact stress in a human hip is not uniform and it changes with different body positions. The changing location of the peak contact stress during gait may indicate the predilection sites for further development of osteoarthritis in the hip. On the basis of laboratory measurements and by using mathematical models of forces and stresses in human hip we determined the points of the peak contact stress in successive phases of gait. Results show that the peak stress points are mostly located in the posterior-medial portion of the weight bearing area, which corresponds well to the clinical observations. It is also shown that in the pathological conditions of hip dysplasia the peak contact stress trajectory is located more laterally and anteriorly. PMID- 11005662 TI - G-protein coupled progesterone receptors in the plasma membrane of fungus Rhizopus nigricans. AB - We have demonstrated simultaneous existence of progesterone receptors and GTPase activity in the membranes prepared from the filamentous fungus Rhizopus nigricans. The results obtained with pertussis toxin treated fungal mycelium suggest that these receptors do not couple to Gi-Go-proteins and play a role in the induction of steroid hydroxylating enzyme system by steroid substrates in the fungus. PMID- 11005663 TI - The effect of lamina propria on the growth and differentiation of urothelial cells in vitro. AB - The effect of lamina propria on the growth and differentiation of the mouse urinary bladder urothelial cells in vitro has been studied by light and electron microscopy using morphological and immunohistochemical methods. Three different types of urothelial cultures were maintained on a porous membrane in serum-free medium for 10 days. Our results showed that urothelial cells in culture types I and II were organised as the multilayer epithelium showing terminal differentiation, from the porous membrane toward the surface. On the surface of culture type I and II, superficial urothelial cells were highly differentiated. They formed tight junctions and had plaques of an asymmetric unit membrane, a hallmark of terminal differentiation in bladder superficial cells. These cultured superficial cells were also cytokeratin 20 positive. On the contrary, urothelial cells in culture type III have shown a limited capacity for survival and have grown as poorly differentiated monolayer. These results revealed that reciprocal intercellular signalling between the lamina propria and urothelial cells is essential to achieve terminal differentiation of urothelial cells in vitro. PMID- 11005664 TI - Mouse urothelial cells in early postnatal development--proliferation and apical plasma membrane specialization. AB - The purpose of this work was to investigate proliferation and differentiation of the mouse urothelial cells from the day of birth until the 5th postnatal day. The expression of proliferating cell nuclear antigen (PCNA) was studied immunocytochemically and the differentiation of apical plasma membrane of superficial cells was analysed by scanning microscopy. It was established that proliferation activity is very high during all five days since PCNA positive cells are seen in the superficial and in basal cell layer of the urothelium. Results of scanning microscopy show that the differentiation of superficial urothelial cells is a nonsynchronous process, which gradually progresses from the day of birth and leads into homogeneous population of terminally differentiated superficial cells on the 5th postnatal day. PMID- 11005665 TI - Reticulocyte analysis using light microscopy and two different flow cytometric procedures. AB - The reticulocyte count is a clinically important indirect indicator of erythropoietic activity of the bone marrow. Reticulocyte enumeration by light microscopy is rather inaccurate and has poor reproducibility. Automation of the reticulocyte count by means of flow cytometry has considerably improved the quality of this investigation. In our study, we compared three methods of establishing the blood reticulocyte number: the microscopic brilliant cresyl blue method and two flow cytometric procedures using thiazole orange (TO), namely FACSort (Becton-Dickinson) and EPICS Profile (Coulter). The aims of the study were (1) to select the most suitable TO concentration to be used with the EPICS Profile cytometer, (2) to determine the correlation between the microscopic method and the two flow cytometric procedures, and (3) to appraise the suitability of flow cytometry for reticulocyte analysis in routine clinical work. According to our results, the most appropriate TO concentration for the EPICS Profile counter is 0.1 mg/L. We observed a good correlation between the three methods tested; the correlation coefficients ranged from 0.82 to 0.87. The mean intra-assay coefficients of variation for the microscopic method and the EPICS Profile and FACSort procedures were 27.5%, 8.4% and 6.3%, respectively. PMID- 11005666 TI - The contribution of HLA-Class II antigens in humoral non-response and delayed response to HBsAG vaccination. AB - The variability in the immune response modulated by HLA alleles may be an important factor for the induction of the protective effect of HBsAg vaccines. We present here the analysis of HLA-DRB1, DQB1 and DQA1 alleles and their combinations in the group of 36 individuals with poor humoral immmune response to HBsAg vaccination. Comparison with the control group, consisted of 60 randomly choosen healthy subjects, revealed that the DRB1*1601, DQB1*0502, DQA1*0102 haplotype is overrepresented in the group of hyporesponders and may therefore be regarded as a factor influencing poor antibody responsiveness. We observed that after revaccination two of three individuals who failed to develop anti-HBs antibodies carry the same phenotype DRB1*0101,DRB1*0301;DQB1*0501,DQB1*0201;DQA1+ ++*0101,DQA1*0501, which supports the conjecture that immunogenicity of the HBsAg vaccine depends on specific combination of HLA DR and DQ molecules on antigen presenting cells. PMID- 11005667 TI - Hypomethylation of alphoid DNA and classical satellite DNA on chromosome 1, 9, 16 and Y in extraembryonic tissue. AB - The pattern of DNA methylation can be analyzed on methaphase chromosomes with fluorescein labeled antibodies against 5-methylcytosine. In human extraembryonic tissue lower overall intensity of immunofluorescence in centromeric chromosomal regions correspond to hypomethylation of the DNA when compared with normal human lymphocytes. Pericentromeric regions on chromosomes 1,9,16 and heterochromatin on chromosome Y, which reveal lower levels of immunofluorescence, are rich in classical satellite DNA type II and III. In our experiment methylation-sensitive restriction enzymes, alphoid and classical satellite DNA probes specific for chromosomes 1,9,16 and Y were used. Southern blot analysis on cells from extraembryonic tissue revealed different extent of hypomethylation in different chromosomal regions. Our results confirm overall and sequence-specific hypomethylation of DNA in cells from extraembryonic tissue in comparison with somatic cells. PMID- 11005668 TI - Effect of pH on red blood cell deformability. AB - The effect of pH on the red blood cell (RBC) deformability, which is a consequence of a change of cell membrane elastic properties is studied experimentally. With the intention to reduce the effects on deformability of cell geometry and cytoplasmic viscosity, we measured the deformability of the cells with the same volume at various pH of cell suspension from 6.2 to 8.0. Constant cell volume was achieved by varying osmolarity. Deformability was quantified by measuring the elongation of RBCs subjected to velocity gradient in a transparent cone-plate rheoscope. Observed significant decrease of deformability at lower pH leads to the conclusion that membrane elastic properties could be affected by pH changes in the range from 6.2 to 8.0. PMID- 11005669 TI - Weak D and partial D in Slovenian population through serology and genotyping. AB - Weak D red cell phenotype (formerly D(II)) exhibits weaker serological reaction with anti-D antibodies. Weak D occurs in 0.2% to 1% of whites and is caused by qualitatively altered RhD proteins called partial D or normal, only weakly expressed RhD proteins that are called weak D. Partial D genes are hybrid alleles between RHD an RHCE genes. 23 partial RHD alleles are described. Weak D phenotypes with reduced expression are likely to possess the normal RHD gene, but the latest findings indicate that weak D alleles carry at least one point mutation. The aim of the present work was to answer an important question how to approach partial and weak D identification in diagnostic use and if it is possible to distinguish between partial D and weak D using commercially available anti-D reagents for routine use. We also wanted to evaluate D-screen kit for partial D identification. We compared phenotypes identified by serological testing and genotypes identified by RHD Multiplex PCR and D(VII) specific ASPA PCR. Our results showed that it is not possible to distinguish between partial and weak D using commercially available anti-D reagents for routine use. D-screen proved to be useful for D(VI) and D(VII) identification, whereas for partial D(DFR) identification we must look for another set of monoclonal antibodies or simply use genotyping methods. In 44 samples with not interpretable serological results out of 80 we found all RHD specific exons present and we classified the samples as weak D. Fourteen types of weak D with at least one point mutation were recently proposed. Designing of allele specific PCRs for identification of proposed types of weak D is in progress. PMID- 11005670 TI - Determining the parameters of concentration-response relationship for histamine on the isolated portion of guinea pig stomach and guinea pig right atrium. AB - In our previous experiments measuring acid secretion in the isolated mouse stomach we found higher EC50 values (agonist concentration, evoking half maximal effect) for histamine than in the experiments on the isolated guinea pig right atrium (contraction frequency). The difference could be due to nonequilibrium conditions in the isolated mouse stomach preparation or to species differences. In order to check this possibility and to test the consistency of concentration response relationship (CRR) parameter determination on an isolated secretory organ, we compared EC50 for histamine and pA2 for famotidine on the two isolated preparations from the same species, guinea pigs. Both, EC50 and pA2 values were significantly different when measuring gastric acid secretion (9.7 microM and 6.07) from the parameters, found in the isolated right atrium measuring frequency of contractions (1.6 microM and 7.06). From these results it can be concluded that the isolated portion of guinea pig stomach is suitable for CRR determination; but the parameters obtained are in the range of higher concentrations than the real ones because of kinetic particularities of the preparation. PMID- 11005671 TI - The oxygen uptake threshold during incremental exercise test. AB - The linear relationship between oxygen consumption (VO2) and exercise intensity is a well established phenomenon observed during incremental exercise. Recently, a non-linear increase in VO2 has been reported by Zoladz et al., who used a relatively complicated method to describe the phenomenon. In this study, we tried to ascertain whether the same phenomenon, which we named the oxygen uptake threshold (OUT), could be described by a simple method, using the two best fitting lines adopted for the less and more steep parts of the VO2 increase. Our hypothesis was that the non-linear VO2 increase was the result of a continuous VO2 increase (oxygen drift) occurring during the more intense steps only. Therefore, we analysed the VO2 time course during each step. Six cyclists performed an incremental exercise test on a cyclo-ergometer. The lactate threshold (LT) was calculated by using the intersection point of the two best fitting lines in the diagram of log LA (lactate concentration) dependence on log P (Power). The time course of VO2 during each step was analysed by an exponential rise to the maximum model. The results showed that OUT could be determined in five of the six subjects, whereas LT could be determined in all six subjects. The power output determined by OUT (168 +/- 13 W) was similar to that determined by LT (180 +/- 25 W). The VO2 time course during each step showed steady values during low intensity exercise. At intensities above LT and OUT, however, VO2 increased continuously, showing oxygen drift. It may be concluded that OUT is a realistic phenomenon, which is based on oxygen drift. PMID- 11005672 TI - Evaluation of biomechanical status of dysplastic human hips. AB - The evaluation of the biomechanical status of human hip joint was performed. Required parameters were assessed from standard antero-posterior rentgenographs and the value of contact stress on the weight bearing area is calculated. We analysed 45 dysplastic human hips and compared measured values to 81 healthy hips. The peak contact stress on the weight bearing area is considerably and statistically significantly higher in dysplastic hips compared to healthy hips. The Wiberg centre-edge angle is statistically significantly smaller in group of dysplastic hips compared to healthy hips. PMID- 11005673 TI - The influence of the successive depolymerization and polymerization of cytoskeleton components on the fibroblast shapes. AB - Monitoring the influence of the cytoskeleton polymers on the shape of fibroblasts, performing the experiments of repeated degradation and polymerization of microtubules and microfilaments, we found out that the presence of microtubules is necessary in order to regenerate the proper functional structure of microfilaments, and vice versa. PMID- 11005674 TI - Comparative toxicity of trivalent and pentavalent inorganic and methylated arsenicals in rat and human cells. AB - Biomethylation is considered a major detoxification pathway for inorganic arsenicals (iAs). According to the postulated metabolic scheme, the methylation of iAs yields methylated metabolites in which arsenic is present in both pentavalent and trivalent forms. Pentavalent mono- and dimethylated arsenicals are less acutely toxic than iAs. However, little is known about the toxicity of trivalent methylated species. In the work reported here the toxicities of iAs and trivalent and pentavalent methylated arsenicals were examined in cultured human cells derived from tissues that are considered a major site for iAs methylation (liver) or targets for carcinogenic effects associated with exposure to iAs (skin, urinary bladder, and lung). To characterize the role of methylation in the protection against toxicity of arsenicals, the capacities of cells to produce methylated metabolites were also examined. In addition to human cells, primary rat hepatocytes were used as methylating controls. Among the arsenicals examined, trivalent monomethylated species were the most cytotoxic in all cell types. Trivalent dimethylated arsenicals were at least as cytotoxic as trivalent iAs (arsenite) for most cell types. Pentavalent arsenicals were significantly less cytotoxic than their trivalent analogs. Among the cell types examined, primary rat hepatocytes exhibited the greatest methylation capacity for iAs followed by primary human hepatocytes, epidermal keratinocytes, and bronchial epithelial cells. Cells derived from human bladder did not methylate iAs. There was no apparent correlation between susceptibility of cells to arsenic toxicity and their capacity to methylate iAs. These results suggest that (1) trivalent methylated arsenicals, intermediary products of arsenic methylation, may significantly contribute to the adverse effects associated with exposure to iAs, and (2) high methylation capacity does not protect cells from the acute toxicity of trivalent arsenicals. PMID- 11005676 TI - Prediction of in vivo metabolic clearance of 25 different petroleum hydrocarbons by a rat liver head-space technique. AB - In vitro rates of metabolism and Michaelis-Menten constants were determined for 25 different C6 to C10 hydrocarbons using rat liver slices in a vial head-space equilibration system. The rates of metabolism were compared with steady-state levels obtained in vivo in the same strains of rats after inhalation. Aromates were metabolized at a higher rate than naphthenes n-alkanes, isoalkanes and 1 alkenes. The aromates showed, in contrast to the other hydrocarbons investigated, increased metabolism with increasing number of carbon atoms up to C8 (o-xylene, the most extensively metabolized compound). The in vivo steady-state concentrations of the aromates in blood were inversely related to the in vitro efficiency of their metabolism. This explains the pattern of blood levels observed for the C6 to C10 aromates in the rat after inhalation, with o-xylene demonstrating the lowest concentration. In general, the extent of tissue metabolism of the investigated hydrocarbons might be of greater importance for their body distribution than their lipophilicity, especially for the highly metabolized compounds. The high in vitro intrinsic liver clearances found for the aromates indicate a flow-dependent metabolism of these hydrocarbons in vivo. The head-space liver slice equilibration system seems to work adequately for metabolic studies of hydrocarbons with different volatility and water solubility. PMID- 11005675 TI - Effects of a cisplatin-chondroitin sulfate A complex in reducing the nephrotoxicity of cisplatin. AB - To assess the effects of a macromolecular prodrug in reducing the nephrotoxicity of cisplatin (CDDP), chondroitin sulfate A (CSA) with a mean molecular weight of 23,000 Da was used to form a complex with CDDP, and the pharmacokinetics and toxicology of the resulting complex were examined in rats in comparison with those of CDDP. The total plasma platinum levels and urinary accumulation were determined up to 3 h following a bolus injection of 2 mg/kg. The results of the pharmacokinetic analysis showed that the complex suppressed the rapid distribution of CDDP, decreased the renal clearance and resulted in over fivefold higher AUC values within 3 h in comparison with CDDP treatment. In addition, the plasma levels of the drug following administration of the complex decreased greatly with time throughout the experimental period (3-24 h), whereas a slow elimination was observed following CDDP administration, which was due to the irreversible protein binding of CDDP. The tissue-to-plasma partition ratio at 10 min also indicated that the CDDP-CSA complex controlled the perfusion of CDDP to tissues, especially to the kidney. The accumulation in various tissues was evaluated at 3 h and 24 h following the injection of 5 mg/kg. Marked differences in renal accumulation were found within 3 h. Significant reductions in accumulation in the kidney, lung, muscle and whole blood were found within 24 h of administration of the complex. The renal toxicity of the CDDP-CSA complex was evaluated by measuring blood urea nitrogen (BUN), serum creatinine (Cr) and the ratio of terminal kidney weight to body weight at doses of 2 mg/kg and 5 mg/kg. The complex displayed a much lower nephrotoxicity at 5 mg/kg in comparison to CDDP, and similar results were obtained at 2 mg/kg. This suggests that the complex changed the toxicodynamics of CDDP. Moreover, the anticancer activity of the CDDP-CSA complex, tested against SW 4800 human colon cancer cells and HeLa human cervix cancer cells in vitro, showed no decrease as compared with that of free CDDP. We conclude that the CDDP-CSA complex had the same activity as the parent drug but showed reduced nephrotoxicity at high doses of CDDP through an improvement in the pharmacokinetics of CDDP, which resulted from both the minimization of entry into normal tissues and renal clearance. In addition, it is also possible that different intracellular interactions in renal cells play a role in protection against the nephrotoxicity of high doses of CDDP. PMID- 11005677 TI - Invasion and distribution of methanol. AB - After the enzyme systems responsible for methanol oxidation were blocked by ethanol, five test persons were given methanol at a dose of approximately 10 mg/kg weight, once orally and once parenterally. Taking into account the endogenous blood methanol levels detectable before the administration of methanol, C0 concentrations of 11.1-15.9 mg/kg were reached. This corresponds to a distribution volume of approximately 0.77 +/- 0.07 l/kg, which is comparable to the 0.78 +/- 0.09 l/kg obtained for ethanol. After parenterally administering methanol as a bolus, the distribution half-life was on average 8 min (range: 3.8 13.8 min). After oral administration of methanol diluted in 100 ml water on an empty stomach, invasion took place with a half-life of approximately 5 min (3.8 6.9 min). In one case, however, due to vegetative disturbances the invasion half life was 23.1 min. PMID- 11005679 TI - Induction of N-nitrosodimethylamine metabolism in liver and lung by in vivo pyridine treatments of rabbits. AB - N-Nitrosodimethylamine is a procarcinogen that is activated by cytochrome P450 dependent N-nitrosodimethylamine N-demethylase to labile alpha-carbon hydroxylated products further resulting in active methylating agents. In vivo intraperitoneal administration of pyridine to rabbits significantly increased N nitrosodimethylamine N-demethylase activity by 6.9- and 5.2-fold in liver and lung microsomes, respectively. Although, p-nitrophenol hydroxylase and aniline 4 hydroxylase activities were markedly enhanced by pyridine treatment in liver about 4.4- and 5.8-fold, respectively, no change was observed in the activities of these enzymes in lung microsomes. Pyridine treatment also elevated P450 contents of liver and lung by 2.04- and 1.4-fold, respectively. SDS-PAGE of pyridine-induced liver microsomes revealed a protein band of enhanced intensity having Mr of 51,000 migrating in the region of cytochrome P4502E1. The results obtained in this study demonstrated for the first time, a significant 5.2-fold induction of NDMA N-demethylase activity in the rabbit lung over the controls. Pyridine is readily absorbed by inhalation and is a constituent of tobacco and tobacco smoke. Thus induction of NDMA N-demethylase suggests that in the lung, as in the liver, pyridine may stimulate the metabolic activation of this nitrosamine significantly. PMID- 11005678 TI - Association of azinphos-methyl with rat erythrocytes and hemoglobin. AB - To investigate whether hemoglobin might serve as a biomarker of exposure to azinphos-methyl (AZM) encountered by agricultural workers, we exposed rats to [14C]azinphos-methyl ([14C]AZM). We administered single doses of 1.5 mg/kg, 3 mg/kg, or 6 mg/kg of [14C]AZM by gavage to rats and collected blood 3 days later. We found a dose-dependent association between radioactivity and erythrocytes and hemoglobin (measured by liquid scintillation spectrometry). In another experiment, we administered a single dose of 3 mg/kg [14C]AZM by gavage to rats and collected blood 3, 11, 15, and 22 days after administration. Radioactivity continued to be associated with erythrocytes and hemoglobin at all time-points. Brain and plasma acetylcholinesterase (AChE) activities were not significantly inhibited, nor did we observe signs of acute toxicity in any of the treated animals. Loss of radioactivity associated with erythrocytes and hemoglobin over the study period was consistent with the expected kinetics of erythrocyte turnover, indicating adduct stability. Approximately 0.49%, 0.43%, 0.39%, and 0.32% of the original radioactivity was recovered in the hemoglobin 3 11, 15, and 22 days, respectively, after administration. These data support our hypothesis that AZM may form a hemoglobin adduct and may be useful as a biomarker of AZM exposure. PMID- 11005680 TI - CYP1A-dependent activation of xenobiotics in endothelial linings of the chorioallantoic membrane (CAM) in birds. AB - Metabolic activation of the heterocyclic amine 3-amino -1,4-dimethyl-5 H pyrido[4,3-b]indole (Trp-P-1) and 7-ethoxyresorufin O-deethylase (EROD) activity were examined in the chorioallantoic membrane (CAM) of 15-day-old chicken and 18 day-old eider duck embryos. The embryos were pretreated with an Ah receptor agonist, i.e. beta-naphthoflavone (BNF) or 3,3',4,4',5-pentachlorobiphenyl (PCB 126), or vehicle in ovo. BNF and PCB 126 induced EROD activity and covalent binding of [3H]Trp-P-1 seven- to tenfold in the CAM of chicken embryos. In the CAM of eider duck embryos, which are known to be nonresponsive to coplanar PCBs, PCB 126 treatment had no effect on EROD activity or covalent binding of [3H]Trp-P 1 whereas BNF treatment increased these activities five- and threefold, respectively. Light microscopic autoradiography was used to identify the cellular localization of covalent binding of [3H]Trp-P-1 in the CAM. Preferential binding was observed in endothelial cells in intraepithelial capillaries in the chorionic epithelium and in blood vessels in the mesenchymal layer. The addition of the CYP1A inhibitor ellipticine abolished the covalent binding of [3H]Trp-P-1 in the CAM of BNF- and PCB 126-treated chicken and eider duck embryos. The results suggest that CYP1A-dependent metabolic activity can be induced in blood vessel endothelia in the CAM of bird embryos following exposure to Ah receptor agonists and that the CAM may be a target tissue for CYP1A-activated environmental pollutants. Furthermore, the highly vascularized CAM could be used as a model for studies of Ah receptor-mediated alterations in the vasculature. PMID- 11005681 TI - Polymorphism of the N-acetyltransferase (NAT2), smoking and the potential risk of periodontal disease. AB - Periodontal disease is a common multifactorial process that leads to bone destruction and tooth loss. Interactions of environmental and genetic factors determine the extent and severity of periodontal disease. Smoking is one of the risk factors for periodontal disease, and the risk may be influenced by the polymorphism of N-acetyltransferase (NAT2) via metabolism of smoke-derived xenobiotics. We therefore hypothesized that a NAT2 genotype would be a risk factor for periodontal disease. A total of 154 Caucasian subjects were assigned to one of two groups (1) no or mild and (2) severe periodontal disease based on radiographic (bone destruction) and clinical criteria (probing depth, attachment loss) and the number of teeth. In all subjects genotyping for mutations on NAT2 was performed by means of PCR and RFLP analysis. In the less-affected group genotyping showed a fraction of predicted slow and rapid acetylators (53.6% and 46.4%, respectively) corresponding to the normal distribution in Caucasians. Severely affected patients were predominantly slow acetylators, the odds ratios being between 2.38 and 5.02 for the NAT2-related risk depending on the outcome parameters chosen. Adjustment for age had no influence on these findings. Our data indicate that the slow acetylator phenotype is associated with a higher risk of periodontitis, especially with respect to the severity of the disease. Possible implications with respect to the risk associated with smoking are discussed. PMID- 11005682 TI - Statistics and clinical oncology. PMID- 11005683 TI - Committee on publication ethics (COPE): guidelines on good publication practice. PMID- 11005684 TI - Randomized trials in early prostate cancer. I: Requiem or renaissance? PMID- 11005685 TI - Randomized trials in early prostate cancer. II: hormone therapy and radiotherapy for locally advanced disease: a question is still unanswered. MRC PR07 Trial Management Group. PMID- 11005686 TI - Complications of treatment with local field external beam radiotherapy for localized prostate cancer. AB - We performed this analysis to document the rate and severity of complications both during and after local field irradiation for localized prostate cancer and to assess the influence of three-dimensional conformal radiation therapy (3DCRT) on these complications. Bowel, urinary and cutaneous toxicities were noted in all patients treated with primary radiotherapy (RT) for prostate cancer using Radiation Therapy Oncology Group/European Organization for Research on Treatment of Cancer scores. Evaluations were performed weekly during RT, 6 weeks after RT and 3 monthly thereafter. Data on 111 were analysed. We also compared the complications of conventional and 3DCRT. The serious complication rates identified in this study compare favourably with those reported in the literature. Only 3.6%, 0.8% and 0% had acute grade 3 or 4 urinary, bowel or skin reactions respectively. Only two patients had chronic grade 3 or 4 urinary complications. Thus far no patients have developed chronic bowel or skin complications greater than grade 2. Despite dose escalation from 66 Gy to 70 Gy (with 3DCRT), there was a trend towards reduced toxicity when 3DCRT was compared with conventional radiation. RT is a well-tolerated treatment for early stage carcinoma of the prostate and our complication rates are compatible with international experiences. Further follow-up is required to determine the efficacy of treatment, the incidence of impotence and the final number of late complications. PMID- 11005687 TI - Radiation therapy in localized prostate cancer: long-term results and late toxicity. AB - This study was performed to determine the long-term results of radiotherapy in localized prostate cancer. The frequency and severity of late gastrointestinal and genitourinary toxicities were also investigated. Between 1980 and 1991, 206 men with localized prostate cancer were treated with radiotherapy alone at our hospital. Biopsy, transurethral resection of the prostate, or both, confirmed the diagnosis. Overall survival, cancer-specific survival, disease-free survival, local recurrence-free survival and metastasis-free survival were determined and compared with age, stage, grade and diagnostic intervention. A change in treatment policy occurred in 1987. The treatment results of the two periods are compared. Late toxicity was registered according to the Radiation Therapy Oncology Group scale. The 8-year overall survival rates were 60%, 44% and 29% for Stages T1, T2 and T3 respectively (P=0.028). The 8-year cancer-specific survival rates were 86%, 66% and 34% for Stages T1, T2 and T3 respectively (P=0.002). Transurethral resection of the prostate had a negative influence on cancer-free and disease-free survivals, owing especially to an increased incidence of metastases. In the bladder and rectum, late toxicity of grade 2 or more occurred in 26 (13%) of the 199 patients who were alive at 6 months after radiotherapy. One of these patients developed late toxicity 5 years after the radiotherapy. Radiation dose and field size had no detectable influence on the development of late toxicity. The difference between overall survival and cancer-specific survival confirms that many of these patients die without any clinical signs of prostate cancer or metastases. This observation puts a question mark over whether a biochemical endpoint alone is of clinical relevance. Although not statistically significant, local recurrence-free, disease-free and cancer-specific free survivals from 1987 onwards improved for larger tumours, which was most likely due to the administration of higher doses of radiation. PMID- 11005688 TI - Seminoma metastatic to the prostate resulting in a rectovesical fistula. AB - We report an unusual presentation of metastatic seminoma within the prostate gland. Histological diagnosis was obtained using trans-rectal ultrasound guided prostatic biopsy. The patient developed a rectovesical fistula after ten weeks of chemotherapy, which healed following a complete radiological response to treatment PMID- 11005689 TI - Rare cancers: how should we organize expertise? PMID- 11005690 TI - Radiation-induced malignant melanoma of the cervix. AB - We report the occurrence of a malignant melanoma of the uterine cervix in a patient previously irradiated for a squamous cell carcinoma of the cervix. The management of malignant melanoma of the cervix is reviewed. The risk of second cancers in general, malignant melanoma in particular, after radiotherapy for cervical cancer is also discussed. PMID- 11005691 TI - Extraosseous osteosarcoma of the penis. AB - Osteosarcomas have been reported as arising in a number of extraosseous primary sites, most commonly in the retroperitoneum and the muscles of the thighs and limb girdles, but also in a variety of other organs. We present a case arising in the penis, which we believe to be only the fifth reported in the literature and the only documented long-term survivor. Careful histopathological analysis and surgical management remain of key importance in the management of all sarcomas arising in soft tissue. PMID- 11005692 TI - Oral fluoropyrimidines in the treatment of colorectal cancer. PMID- 11005693 TI - Radiotherapy is a tolerable and potentially useful therapy in gastric cancer. AB - The role of radiotherapy in gastric cancer remains controversial. Most previous clinical trials have not shown any convincing benefit. However, current imaging is probably sufficiently sophisticated to allow the stomach to be treated adequately with radiotherapy. Three case histories are presented, which illustrate the use of radiotherapy in both palliative and radical settings. PMID- 11005694 TI - Antitumour activity of heparin on metastatic colon cancer. AB - We present the case history of a 47-year-old patient with metastatic colon cancer, whose anticoagulant treatment led to a prolonged disease-free survival. PMID- 11005695 TI - Phase I dose escalation study of gemcitabine and paclitaxel plus colony stimulating factors in previously treated patients with advanced breast and ovarian cancer. AB - Gemcitabine and paclitaxel (PTX) are among the most active new drugs in advanced breast and ovarian cancer. In this Phase I study, we used fixed doses of gemcitabine administered on days 1 and 8 and escalating doses of paclitaxel on day 1 of a 21-day cycle in patients with pretreated metastatic breast or ovarian cancer. The dose of gemcitabine was fixed at 1,000 mg/m2; PTX was commenced in the first small patient group at a dose of 90 mg/m2, which was then escalated in subsequent groups by 30 mg/m2 per step. From the third dose level onwards, all patients received granulocyte colony-stimulating factor 300 microg by subcutaneous injection on days 5 and 6, and granulocyte macrophage colony stimulating factor on days 15-18. Cohorts of at least 3 patients were treated at each dose level. Dose escalation was stopped if at least a third of the patients in a given cohort had dose-limiting toxicity (DLT), which was defined as grade 4 neutropenia or thrombocytopenia, or grade 3-4 non-haematological toxicity. The maximum tolerated dose (MTD) was defined as the dose level immediately below that causing DLT in one-third of the patients or more. Evaluation of the tumour response was performed every three cycles. Forty-five patients (31 with breast cancer, 14 with ovarian cancer) were treated at seven different dose levels. Only at the seventh PTX dose level was DLT observed after the first course of therapy: three grade 4 neutropenia, one grade 4 thrombocytopenia, and one grade 4 anaemia. DLT occurred in 5/6 patients at at PTX dose of 270 mg/m2; therefore dose escalation was stopped at that level and the dose immediately before it (PTX 240 mg/m2) was considered as the MTD and recommended for further studies. No toxic deaths occurred. Grade 3-4 uncomplicated neutropenia was observed in four patients. Three had uncomplicated grade 3-4 thrombocytopenia. One patient had grade 3 and one grade 4 anaemia. Nonhaematological side effects were generally mild. Among 30 evaluable patients with metastatic breast cancer, four complete responses (CR) (13%) and 12 partial responses (PR) (40%) were observed, for an overall response rate of 53% (95% confidence interval (CI) 34-72). The median duration of response was 31 weeks. Among 13 evaluable patients with advanced ovarian cancer, one CR (8%) and five PRs (38%) were observed, for an overall response rate of 46% (95% CI 19-78). The median duration of response was 32 weeks. Our study shows that gemcitabine and PTX can be administered in combination in patients with breast and ovarian cancer without unexpected toxicities and with encouraging therapeutic results. PMID- 11005696 TI - A multicentre phase II pilot study of epirubicin and Taxol (paclitaxel) in patients with advanced breast cancer. AB - Anthracyclines are the gold standard monotherapy for metastatic breast cancer. Higher response rates are seen with drug combinations, especially with newer agents such as taxanes. The purpose of this study was to evaluate the toxicity and activity of the combination of paclitaxel and epirubicin in patients with advanced breast cancer. Thirty-five women with locally advanced or metastatic breast cancer (first and second relapse) were treated with epirubicin 75 mg/m2 and paclitaxel 200 mg/m2 3-weekly. Six centres recruited 35 patients; 34 (97%) were assessable for response. Eighteen had undergone prior chemotherapy, including six (17%) with anthracycline-containing regimens. Grade 4 neutropenia was found in 33 patients (94%), which was of 4 days' average duration; however, infective complications were rare, with only nine cycles (6%) complicated by neutropenic sepsis. There were two sepsis-related deaths. Symptomatic cardiotoxicity was infrequent, although a >15% decline in cardiac function was recorded in five patients (14%). Grade 3 peripheral neuropathy occurred in three patients (9%). The overall response rate was 50% (95% confidence interval 33-67) (complete response 12%; partial response 38%), with a median duration of response of 31 weeks. The median time to progression was 27 weeks, with a median survival of 48 weeks. This regimen appears to be a relatively safe, tolerable and effective treatment for advanced breast cancer. A United Kingdom Co-ordinating Committee for Cancer Research Phase III trial (AB-01) comparing this combination of epirubicin and paclitaxel with cyclophosphamide and paclitaxel completed accrual in November 1999. PMID- 11005697 TI - Unresectable adenoid cystic carcinoma of the trachea treated with chemoradiotherapy. PMID- 11005698 TI - Docetaxel-induced radiation recall dermatitis and successful rechallenge without recurrence. PMID- 11005699 TI - Evaluation of a nurse-led telephone clinic in the follow-up of patients with prostate cancer. PMID- 11005700 TI - Metastatic breast cancer presenting with uveitis. PMID- 11005701 TI - Is generic prescribing acceptable in epilepsy? AB - There is considerable debate about the role of generic prescribing for people with epilepsy. The arguments go beyond simple considerations of cost on one hand and the possibility of toxicity or loss of seizure control on the other. The concepts of bioavailability and bioequivalence require further consideration. The measures that are currently used may not apply equally well to all situations. For example, additional measures may be needed for controlled-release preparations and in the other special cases. There is an extensive literature on the bioequivalence of various phenytoin preparations. This anticonvulsant drug is poorly soluble in water, has nonlinear kinetics and has a narrow therapeutic range, implying that problems with bioequivalence are likely to occur. This is borne out by clinical experience. There are a few published investigations on carbamazepine. The systematic studies, on the whole, fail to show major differences in bioequivalence between the various formulations. There is sparse information on the comparison between generic and proprietary formulations of other anticonvulsant drugs. Whatever arguments might be put forward supporting brand name or generic prescribing, there are strong reasons for recommending tight control on the consistency of anticonvulsant drugs, both generic and proprietary. There is also a strong case for ensuring that the physician who signs the prescription remains in control of the situation and that any decisions that the physician makes should be based on accurate and reliable information. PMID- 11005702 TI - A risk-benefit assessment of risperidone for the treatment of behavioural and psychological symptoms in dementia. AB - The importance of behavioural and psychological symptoms in dementia (BPSD) is increasingly being recognised. Symptoms such as verbal and physical aggression, agitation, sleep disturbances and wandering are common, cause great distress to caregivers and are likely to lead to institutionalisation of patients. At present, these symptoms are also more amenable to treatment compared with the progressive intellectual decline caused by dementing illnesses. The care of individuals with BPSD involves a broad range of psychosocial treatments for the patient and his or her family. If pharmacotherapy is deemed necessary to manage BPSD, a careful balance must be struck between the benefits of symptom control and the inherent risks associated with most psychotropic agents in the elderly. Elderly patients in general, and patients with dementia in particular, are more sensitive to medication adverse effects, including anticholinergic effects, orthostatic hypotension, sedation, parkinsonism, tardive dyskinesia and cognitive impairment than younger patients with dementia or individuals without dementia. To date, treatment of symptoms of aggression and psychosis has relied on the empirical use of antidepressants, anxiolytics, typical antipsychotics (neuroleptics) and other agents. Treatment-limiting adverse effects are frequently reported with all of these agents. However, it is the typical antipsychotics and the atypical antipsychotic clozapine that are associated with the greatest risk of adverse effects in the elderly. The present review highlights the issues that limit the use of older psychotropic agents in the elderly, and presents an assessment of the available evidence concerning the efficacy, safety and tolerability of the atypical antipsychotic risperidone, in the treatment of BPSD in elderly patients with dementia. The extensive clinical development programme for risperidone has shown the drug to be effective and well tolerated in many fragile patients. As a result of its efficacy and safety profile, risperidone can be used for the treatment of behavioural and psychological symptoms in patients with dementia. Risperidone therefore represents a significant addition to the armamentarium for BPSD. While efforts continue in the development of treatment for the cognitive decline associated with dementia, treatment is now available for the noncognitive symptoms. By treating the latter, risperidone has the potential to be of substantial benefit to patients with dementia, their carers and the costs of healthcare. PMID- 11005703 TI - Comparative tolerability of the HMG-CoA reductase inhibitors. AB - The availability of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors has revolutionised the treatment of lipid abnormalities in patients at risk for the development of coronary atherosclerosis. The relatively widespread experience with HMG-CoA therapy has allowed a clear picture to emerge concerning the relative tolerability of these agents. While HMG-CoA reductase inhibitors have been shown to decrease complications from atherosclerosis and to improve total mortality, concern has been raised as to the long term safety of these agents. They came under close scrutiny in early trials because ocular complications had been seen with older inhibitors of cholesterol synthesis. However, extensive evaluation demonstrated no significant adverse alteration of ophthalmological function by the HMG-CoA reductase inhibitors. Extensive experience with the potential adverse effect of the HMG-CoA reductase inhibitors on hepatic function has accumulated. The effect on hepatic function for the various HMG-CoA reductase inhibitors is roughly dose-related and 1 to 3% of patients experience an increase in hepatic enzyme levels. The majority of liver abnormalities occur within the first 3 months of therapy and require monitoring. Rhabdomyolysis is an uncommon syndrome and occurs in approximately 0.1% of patients who receive HMG-CoA reductase inhibitor monotherapy. However, the incidence is increased when HMG-CoA reductase inhibitors are used in combination with agents that share a common metabolic path. The role of the cytochrome P450 (CYP) enzyme system in drug-drug interactions involving HMG-CoA reductase inhibitors has been extensively studied. Atorvastatin, cerivastatin, lovastatin and simvastatin are predominantly metabolised by the CYP3A4 isozyme. Fluvastatin has several metabolic pathways which involve the CYP enzyme system. Pravastatin is not significantly metabolised by this enzyme and thus has theoretical advantage in combination therapy. The major interactions with HMG-CoA reductase inhibitors in combination therapy involving rhabdomyolysis include fibric acid derivatives, erythromycin, cyclosporin and fluconazole. Additional concern has been raised relative to overzealous lowering of cholesterol which could occur due to the potency of therapy with these agents. Currently, there is no evidence from clinical trials of an increase in cardiovascular or total mortality associated with potent low density lipoprotein reduction. However, a threshold effect had been inferred by retrospective analysis of the Cholesterol and Recurrent Events study utilising pravastatin and the role of aggressive lipid therapy is currently being addressed in several large scale trials. PMID- 11005704 TI - Cardiovascular adverse effects of antipsychotic drugs. AB - Minor cardiovascular adverse effects from antipsychotic drugs are extremely common. They include effects such as postural hypotension and tachycardia due to anticholinergic or alpha1-adrenoceptor blockade, and may occur in the majority of patients at therapeutic dosages. There are a number of pharmacological effects that are of uncertain clinical significance, such as blockade of calmodulin, sodium and calcium channels and alpha2-adrenoceptors in the central nervous system. The most serious consequences of treatment, arrhythmias and sudden death, are probably uncommon and are most likely to be caused primarily by blockade of cardiac potassium channels such as HERG. Incomplete evidence suggests that arrhythmias and sudden death are a particular problem with certain drugs (thioridazine and droperidol), high risk populations (elderly, pre-existing cardiovascular disease, inherited disorders of cardiac ion channels or of antipsychotic drug metabolism) or people taking interacting drugs (such as drugs that prolong the QT interval, e.g. tricyclic antidepressants, drugs that inhibit antipsychotic drug metabolism, or diuretics). Clozapine may be unique in also causing death from myocarditis and cardiomyopathy. Much further research is required to more clearly identify high risk drugs and the populations that are at risk of sudden death, as well as the mechanisms involved and the extent of the risk. PMID- 11005706 TI - Risk classification systems for drug use during pregnancy: are they a reliable source of information? AB - BACKGROUND: In several countries, risk classification systems have been set up to summarise the sparse data on drug safety during pregnancy. However, these have resulted in ambiguous statements that are often difficult to interpret and use with accuracy when counselling patients on drug use in pregnancy. OBJECTIVES: The objective of this study was to compare and analyse the consistency between and the criteria for risk classification for medications used during pregnancy included in 3 widely used international risk classification systems. All 3 systems use categories based on risk factors to summarise the degree to which available clinical information has ruled out the risk to unborn offspring, balanced against the drug's potential benefit to the patient. METHODS: Drugs included in the risk classification systems from the US Food and Drug Administration (FDA), the Australian Drug Evaluation Committee (ADEC) and the Swedish Catalogue of Approved Drugs (FASS), were reviewed and compared on basis of the risk factor category to which they had been assigned. Agreement between the systems was calculated as the number of drugs common to all 3 and assigned to the same risk factor category. In addition, evidence on teratogenicity and adverse effects during pregnancy was retrieved using a MEDLINE search (from 1966 up to 1998) for common drugs classified as teratogenic. RESULTS: Differences in the allocation of drugs to different risk factor categories were found. Risk factor category allocation for 645 drugs classified by the FDA, 446 classified by ADEC and 527 classified by FASS was compared. Only 61 (26%) of the 236 drugs common to all 3 systems were placed in the same risk factor category. Analysis of studies on the safety of common drugs during pregnancy of drugs classified as X by the FDA indicated that the variability in category allocation was not only attributable to the different definitions for the categories, but also depended on how the available scientific literature was handled. CONCLUSIONS: Differences in category allocation for the same drug can be a source of great confusion among users of the classification systems as well as for those who require information regarding risk for drug use during pregnancy, and may limit the usefulness and reliability of risk classification systems. PMID- 11005705 TI - Drug therapy for hyperthyroidism in pregnancy: safety issues for mother and fetus. AB - Hyperthyroidism (thyrotoxicosis) in pregnancy and the child bearing years is usually attributable to Graves' disease. This is an autoimmune condition in which thyroid-stimulating immunoglobulins (TSI) cause hyperthyroidism. As a rule, pregnancy complicates the management of hyperthyroidism, rather than vice versa. However, patients who remain thyrotoxic during pregnancy are at increased risk of maternal and fetal complications, particularly miscarriage and stillbirth. Therefore, bodyweight loss, eye signs and a bruit over the thyroid gland in a pregnant woman warrant thyroid investigation. Investigations should include measurement of serum free thyroid hormone levels [free thyroxine (T4) and free triiodothyronine (T3)] rather than total T4 and T3 levels, because total T4 and T3 levels may be raised in euthyroid pregnancies due to the presence of increased levels of thyroxine binding globulin (TBG). By 20 weeks' gestational age, the fetal thyroid is fully responsive to TSI and to antithyroid drugs. Maternal T4 and T3 and thyrotropin pass across the placenta in small and decreasing amounts as gestation progresses, but thyrotropin releasing hormone, TSI, antithyroid drugs, iodides and beta-blockers are readily transferred to the fetus from the mother. Hyperthyroidism is usually treated throughout pregnancy with an antithyroid drug, preferably propylthiouracil. The smallest dose which controls the disease is given with careful monitoring of free T4 and T3 levels to minimise the risk of fetal hypothyroidism and goitre. Bilateral subtotal thyroidectomy may be an option for a small number of patients with hyperthyroidism in pregnancy. PMID- 11005707 TI - If bone is the answer, then what is the question? AB - In the 19th century, several scientists attempted to relate bone trabecular morphology to its mechanical, load-bearing function. It was suggested that bone architecture was an answer to requirements of optimal stress transfer, pairing maximal strength to minimal weight, according to particular mathematical design rules. Using contemporary methods of analysis, stress transfer in bones was studied and compared with anatomical specimens, from which it was hypothesised that trabecular architecture is associated with stress trajectories. Others focused on the biological processes by which trabecular architectures are formed and on the question of how bone could maintain the relationship between external load and architecture in a variable functional environment. Wilhelm Roux introduced the principle of functional adaptation as a self-organising process based in the tissues. Julius Wolff, anatomist and orthopaedic surgeon, entwined these 3 issues in his book The Law of Bone Remodeling (translation), which set the stage for biomechanical research goals in our day. 'Wolff's Law' is a question rather than a law, asking for the requirements of structural optimisation. In this article, based on finite element analysis (FEA) results of stress transfer in bones, it is argued that it was the wrong question, putting us on the wrong foot. The maximal strength/minimal weight principle does not provide a rationale for architectural formation or adaptation; the similarity between trabecular orientation and stress trajectories is circumstantial, not causal. Based on computer simulations of bone remodelling as a regulatory process, governed by mechanical usage and orchestrated by osteocyte mechanosensitivity, it is shown that Roux's paradigm, conversely, is a realistic proposition. Put in a quantitative regulatory context, it can predict both trabecular formation and adaptation. Hence, trabecular architecture is not an answer to Wolff's question, in the sense of this article's title. There are no mathematical optimisation rules for bone architecture; there is just a biological regulatory process, producing a structure adapted to mechanical demands by the nature of its characteristics, adequate for evolutionary endurance. It is predicted that computer simulation of this process can help us to unravel its secrets. PMID- 11005708 TI - Regression of blood vessels in the ventral velum of Xenopus laevis Daudin during metamorphosis: light microscopic and transmission electron microscopic study. AB - Structural changes of the ventral velum of Xenopus laevis tadpoles from late prometamorphosis (stage 58) to the height of metamorphic climax (stage 62) were examined by light and transmission electron microscopy. Special emphasis was given to the blood vessel regression. Early changes of velar capillaries were formation of luminal and abluminal endothelial cell processes, vacuolation, and cytoplasmic and nuclear chromatin condensation. At the height of metamorphic climax, transmission electron microscopy revealed apoptotic endothelial cells with nuclear condensation and fragmentation, intraluminal bulging of rounded endothelial cells which narrowed or even plugged the capillary, and different stages of endothelial cell detachment ('shedding') into the vessel lumen. These changes explain the 'miniaturisation' of the velar microvascular bed as well as the typical features found in resin-casts of regressing velar vessels which have been observed in a previous scanning electron microscopy study of the ventral velum. PMID- 11005709 TI - Structural and spatial organisation of brain parenchymal vessels in the lizard, Podarcis sicula: a light, transmission and scanning electron microscopy study. AB - The structure and 3-dimensional pattern of the intraparenchymal microvessels in the brain of the lizard, Podarcis sicula, were studied by a combination of light and transmission electron microscopy as well as scanning electron microscopy of vascular corrosion casts. The angioarchitecture pattern consists of narrow hairpin-shaped microvascular loops of different length originating from the meningeal surface. In each loop, descending and ascending vessels are closely apposed to one another throughout their length and are connected by a narrow U shaped terminal loop at their tips. The 2 limbs of the vessel pairs show a slightly different diameter but lack other structural differences. While some paired vessels give rise to a secondary hairpin-shaped loop with 2 possible branching patterns, there are no anastomotic intraparenchymal connections with analogous neighbouring structures. The cerebral vascular pattern of Podarcis sicula resembles that found in a few representatives of other vertebrate classes. All cerebral vessels structurally appear to be capillaries. Also the observations carried out on semithin and thin sections strongly support the capillary loop model in the Podarcis brain vasculature and, in accordance with studies carried out on various vertebrates, the general submicroscopic features of the brain capillary wall suggest the presence of an endothelial type blood-brain barrier. PMID- 11005710 TI - Innervation of the cavernous body of the human efferent tear ducts and function in tear outflow mechanism. AB - The lacrimal sac and nasolacrimal duct are surrounded by a wide cavernous system of veins and arteries comparable to a cavernous body. The present study aimed to demonstrate the ultrastructure of the nervous tissue and the localisation of neuropeptides involved in the innervation of the cavernous body, a topic not previously investigated. Different S-100 protein antisera, neuronal markers (neuron-specific enolase, anti-200 kDa neurofilament), neuropeptides (substance P, neuropeptide Y, calcitonin gene-related peptide, vasoactive intestinal polypeptide) and the neuronal enzyme tyrosine hydroxylase were used to demonstrate the distribution pattern of the nervous tissue. The ultrastructure of the innervating nerve fibres was also examined by means of standard transmission electron microscopy. The cavernous body contained specialised arteries and veins known as barrier arteries, capacitance veins, and throttle veins. Perivascularly, the tissue was rich in myelinated and unmyelinated nerve fibres in a plexus-like network. Small seromucous glands found in the region of the fundus of the lacrimal sac were contacted by nerve fibres forming a plexus around their alveoli. Many nerve fibres were positive for S-100 protein (S 100), neuron specific enolase (NSE), anti-200 kDa neurofilament (RT 97), calcitonin gene related peptide (CGRP), substance P (SP), tyrosine hydroxylase (TH), and neuropeptide Y (NPY). Vasoactive intestinal polypeptide (VIP) immunoreactivity was only demonstrated adjacent to the seromucous glands. Both the density of nerve fibres as well as the presence of various neuropeptides emphasises the neural control of the cavernous body of the human efferent tear ducts. By means of this innervation, the specialised blood vessels permit regulation of blood flow by opening and closing the lumen of the lacrimal passage as effected by the engorgement and subsidence of the cavernous body, at the same time regulating tear outflow. Related functions such as a role in the occurrence of epiphora related to emotional responses are relevant. Moreover, malfunction in the innervation of the cavernous body may lead to disturbances in the tear outflow cycle, ocular congestion or total occlusion of the lacrimal passages. PMID- 11005711 TI - Differential expression of proteoglycan epitopes by ovine intervertebral disc cells. AB - The alginate bead culture system has been utilised by several groups to examine the in vitro proteoglycan (PG) metabolism of chondrocytes and intervertebral disc cells, but the nature of the PGs produced has not been examined in detail. This is largely due to the difficulty of separating the anionically charged sodium alginate support matrix from PGs which are similarly charged. In the present study ovine annulus fibrosus, transitional zone and nucleus pulposus cells were dissociated enzymatically from their respective matrices by sequential digestion with pronase/clostridial collagenase and DNAase and then cultured in alginate beads for 10 d. The beads were solubilised and subjected to DEAE Sepharose CL6B anion exchange chromatography to separate the sodium alginate bead support matrix material quantitatively from the disc cell PGs. The alginate free bead PGs were then subjected to composite agarose polyacrylamide gel electrophoresis to resolve PG populations and the PGs were transferred to nitrocellulose membranes by semidry electroblotting. The PGs were identified by probing the blots with a panel of antibodies to defined PG core protein and glycosaminoglycan side chain epitopes. Alginate beads of disc cells were also embedded in paraffin wax and 4 microm sections cut to immunolocalise decorin, biglycan, versican, and the 7-D-4 PG epitope within the beads. Decorin and biglycan had similar distributions in the beads, being localised on the cell surface whereas versican and the 7-D-4 PG epitope were immunolocalised interterritoriarly. This study is the first to demonstrate that ovine disc cells synthesise versican in alginate bead culture. Furthermore the immunoblotting studies also showed that a proportion of the 7-D-4 PG epitope was colocalised with versican. PMID- 11005712 TI - Dynamic ultrastructure of mouse pulmonary alveoli revealed by an in vivo cryotechnique in combination with freeze-substitution. AB - A morphological approach to cell dynamics is usually difficult, since routine preparative techniques for electron microscopy always induce artifacts due to cessation of the blood supply into organs. An in vivo cryotechnique followed by the freeze-substitution method probably reduces such problems. It was applied for examining the pulmonary alveoli of BALB/c mice in vivo. The following ultrastructural features were revealed. (1) A surfactant layer provided a continuous covering to the alveolar epithelium. (2) Pleural epithelial cells, alveolar cells and endothelial cells contained many small vesicles and pits. In the alveolar epithelium, they were often localised near microtubules. (3) Typical lamellar structures in large alveolar epithelial cells were rarely detected. (4) Circulating erythrocytes with various shapes were observed in branching blood capillaries. (5) A close association between erythrocytes and the endothelium was seen at the peripheral alveolar septum. Such ultrastructural arrangements may be appropriate for the physiological functions of the pulmonary alveoli, such as exchanges of gases or materials in vivo. PMID- 11005714 TI - Anatomical variations in the human paranasal sinus region studied by CT. AB - A precise knowledge of the anatomy of the paranasal sinuses is essential for the clinician. Conventional radiology does not permit a detailed study of the nasal cavity and paranasal sinuses, and has now largely been replaced by computerised tomographic (CT) imaging. This gives an applied anatomical view of the region and the anatomical variants that are very often found. The detection of these variants to prevent potential hazards is essential for the use of current of endoscopic surgery on the sinuses. In the present work, we have studied the anatomical variants observed in the nasal fossae and paranasal sinuses in 110 Spanish subjects, using CT in the coronal plane, complemented by horizontal views. We have concentrated on the variants of the nasal septum, middle nasal concha, ethmoid unciform process and ethmoid bulla, together with others of lesser frequency. The population studied showed great anatomical variability, and a high percentage (67%) presented one or more anatomical variants. Discounting agger nasi air cells and asymmetry of both cavities of the sphenoidal sinus, which were present in all our cases, the variations most often observed were, in order, deviation of the nasal septum, the presence of a concha bullosa, bony spurs of the nasal septum and Onodi air cells. PMID- 11005713 TI - The growth promoting effects of bFGF, PD-ECGF and VEGF on cultured postimplantation rat embryos deprived of serum fractions. AB - Serum components in which embryos are cultured in vitro are very important for normal embryonic development. In this study, rat serum was fractionated using Macrosep filters to study the effect of a single growth factor. The fractionated serum, both that containing only material greater than 30 kDa molecular weight (> 30 kDa) and that from which material between 30 kDa and 50 kDa had been removed (< 30 kDa+ > 50 kDa), caused significant embryonic growth retardation. Addition of different concentrations of basic fibroblast growth factor (bFGF, 18 kDa), vascular endothelial growth factor (VEGF, 45 kDa) and platelet-derived endothelial growth factor (PD-ECGF, 45 kDa), to fractionated serum (bFGF to > 30 kDa serum and VEGF or PD-ECGF to < 30 kDa+ > 50 kDa serum) partially restored embryonic growth and development according to a morphological scoring system and protein assay. This restoration was clear by all criteria, as well as in yolk sac vascularisation and heart development. The growth promoting effects of all 3 factors were significant but did not reach the level seen in embryos grown in whole rat serum. The effect of these growth factors was also investigated on anembryonic yolk sac development using a concentration for which maximum whole embryonic growth was seen (128 ng/ml bFGF, 1.6 ng/ml VEGF and 4 ng/ml PD-ECGF), and significant anembryonic yolk sac development was found. These findings suggest that the angiogenic factors may have a growth promoting effect on total embryonic development and vascularisation. PMID- 11005715 TI - Ultrastructure of submucosal glands in human anterior middle nasal turbinates. AB - The abundant glands situated in the lamina propria of the human anterior middle nasal turbinate were complex tubules that consist of serous, seromucous, and mucous cells, either singly or in combination. Serous granules were homogeneously dense, but could have a small lighter core. Seromucous granules had a dense rim and a large compartment of appreciably lighter density. Gradation between serous and seromucous granules made precise identification of these secretory cell types difficult. Mucous cells were of conventional morphology. The secretory tubules, which possessed a complement of myoepithelial cells, gradually transformed into ducts or the changeover was relatively sudden. The ductular portions of the tubules consisted either of tall prismatic cells or of shorter columnar cells, both of which lacked secretory granules, but had many mitochondria in their supranuclear cytoplasm. In many cases the ducts, for most of their length, consisted of secretory cells. These glands clearly participate in the elaboration of the glycoconjugate coat that serves to protect the nasal mucosa and keeps it from drying out. PMID- 11005716 TI - The morphometric changes in the gills of the estuarine crab Chasmagnathus granulatus under hyper- and hyporegulation conditions are not caused by proliferation of specialised cells. AB - Chasmagnathus granulatus is a hyper-hyporegulating crab that inhabits changing habitats of salinity in Brazil, Uruguay and Argentina. Since the gills are the main sites for active ion transport in crabs, the adaptive changes in the gill epithelium occurring under different conditions of salinity were studied by means of morphological and morphometric analysis, and immunohistochemical identification of cell proliferation (BrdU technique). In anterior (1-3) gills the epithelium thickness from crabs acclimatised to 12, 34 and 44 g/l ranged from 1.27 to 2.46 microm, with no significant change during acclimatisation, thus denoting a respiratory function. Medial (4-5) gill epithelium was slightly thicker in extreme salinities, but these differences were not statistically significant. In contrast, epithelial thickness of the posterior (6-8) gills increased significantly up to 8.10 microm (dorsal zone of gill 8) both in hyper- and hyposaline media compared with seawater. The dark areas measured in gill 8 treated with AgNO3 revealed putative ion transporting tissue, especially at 12 and 44 g/l, corresponding to the zones of higher epithelial thickness. Hence these areas seem to participate both in hyper- and hyporegulation. Proliferating cells labelled with BrdU almost never occurred in the gills/salinity combinations studied during the initial 48 h of transfer from seawater to hyperconcentrated or diluted media, thus suggesting an increase in cell size rather than cell proliferation. PMID- 11005717 TI - Main trajectories of nerves that traverse and surround the tympanic cavity in the rat. AB - To guide surgery of nerves that traverse and surround the tympanic cavity in the rat, anatomical illustrations are required that are topographically correct. In this study, maps of this area are presented, extending from the superior cervical ganglion to the otic ganglion. They were derived from observations that were made during dissections using a ventral approach. Major blood vessels, bones, transected muscles of the tongue and neck and supra and infrahyoid muscles serve as landmarks in the illustrations. The course of the mandibular, facial, glossopharyngeal, vagus, accessory and hypoglossal nerves with their branches, and components of the sympathetic system, are shown and discussed with reference to data available in the literature. Discrepancies in this literature can be clarified and new data are presented on the trajectories of several nerves. The course of the tympanic nerve was established. This nerve originates from the glossopharyngeal nerve, enters the tympanic cavity, crosses the promontory, passes the tensor tympani muscle dorsally, and continues its route intracranially to the otic ganglion as the lesser petrosal nerve after intersecting with the greater petrosal nerve. Auricular branches of the glossopharyngeal and of the vagus nerve were noted. We also observed a pterygopalatine branch of the internal carotid nerve, that penetrates the tympanic cavity and courses across the promontory. PMID- 11005718 TI - Stereological comparison of 3D spatial relationships involving villi and intervillous pores in human placentas from control and diabetic pregnancies. AB - In human placenta, 3D spatial relationships between villi and the maternal vascular bed determine intervillous porosity and this, in turn, influences haemodynamics and transport. Recently-developed stereological methods were applied in order to examine and quantify these relationships. Placentas were collected after 37 wk from control pregnancies and those associated with maternal diabetes mellitus classified according to duration and severity (White classification scheme). Two principal questions were addressed: (1) are normal spatial arrangements maintained in well-controlled diabetes mellitus? and (2) do arrangements vary between diabetic groups? To answer these questions, tissue sections cut at random positions and orientations were generated by systematic sampling procedures. Volume densities of villi (terminal + intermediate), intervillous spaces and perivillous fibrin-type fibrinoid deposits were estimated by test point counting and converted to global volumes after multiplying by placental volumes. Design-based estimates of the sizes (volume- and surface weighted volumes) of intervillous 'pores' were obtained by measuring the lengths of point- and intersection-sampled intercepts. From these, theoretical numbers of pores were calculated. Model-based estimates (cylinder model) of the hydraulic diameters and lengths of pores were also made. Second-order stereology was used to examine spatial relationships within and between villi and pores and to test whether pair correlation functions deviated from the value expected for 'random' arrangements. Estimated quantities did not differ significantly between diabetic groups but did display some departures from control values in non-insulin dependent (type 2) diabetic placentas. These findings support earlier studies which indicate that essentially normal microscopical morphology is preserved in placentas from diabetic subjects with good glycaemic control. Therefore, it is likely that fetal hypoxia associated with maternal diabetes mellitus is due to metabolic disturbances rather than abnormalities in the quantities or arrangements of maternal vascular spaces. PMID- 11005719 TI - Characterisation of human soft palate muscles with respect to fibre types, myosins and capillary supply. AB - Four human soft palate muscles, and palatopharyngeus, the uvula, the levator and tensor veli palatini were examined using enzyme-histochemical, immunohistochemical and biochemical methods and compared with human limb and facial muscles. Our results showed that each palate muscle had a distinct morphological identity and that they generally shared more similarities with facial than limb muscles. The palatopharyngeus and uvula muscles contained 2 of the highest proportions of type II fibres ever reported for human muscles. In contrast, the levator and tensor veli palatini muscles contained predominantly type I fibres. A fetal myosin heavy chain isoform (MyHC), not usually found in normal adult limb muscles, was present in a small number of fibres in all palate muscles. The mean muscle fibre diameter was smaller than in limb muscles and the individual and intramuscular variability in diameter and shape was considerable. All palate muscles had a high capillary density and an unusually high mitochondrial enzyme activity in the type II fibres, in comparison with limb muscles. No ordinary muscle spindles were observed. The fibre type and MyHC composition indicate that the palatopharyngeus and uvula muscles are functionally involved in quick movements whereas the levator and tensor veli palatini muscles perform slower and more continuous contractions. The high aerobic capacity and the rich capillarisation suggest that the palate muscles are relatively fatigue resistant. Absence of ordinary muscle spindles indicates a special proprioceptive control system. The special morphology of the palate muscles may be partly related to the unique anatomy with only one skeletal insertion, a feature consistent with muscle work at low load and tension and which may influence the cytoarchitecture of these muscles. Other important factors determining the special morphological characteristics might be specific functional requirements, distinct embryological origin and phylogenetic factors. PMID- 11005720 TI - Positional relationships between the masticatory muscles and their innervating nerves with special reference to the lateral pterygoid and the midmedial and discotemporal muscle bundles of temporalis. AB - For an accurate assessment of jaw movement, it is crucial to understand the comprehensive formation of the masticatory muscles with special reference to the relationship to the disc of the temporomandibular joint. Detailed dissection was performed on 26 head halves of 14 Japanese cadavers in order to obtain precise anatomical information of the positional relationships between the masticatory muscles and the branches of the mandibular nerve. After complete removal of the bony elements, the midmedial muscle bundle in all specimens and the discotemporal muscle bundle in 6 specimens, derivatives of the temporalis, which insert into the disc were observed. On the anterior area of the articular capsule and the disc of the temporomandibular joint, the upper head of the lateral pterygoid, the midmedial muscle bundle of temporalis and the discotemporal bundle of temporalis were attached mediolaterally, and in 3 specimens the posterosuperior margin of the zygomaticomandibularis was attached to the anterolateral area of the disc. It is suggested that these muscles and muscle bundles contribute to various mandibular movements. Although various patterns of the positional relationships between the muscles and muscle bundles and the their innervating nerves are observed in the present study, relative positional relationships of the muscles and muscle bundles and of nerves of the mandibular nerve are consistent. A possible scheme of the developmental formation of the masticatory muscles based on the findings of the positional relationships between the muscles and the nerves is presented. PMID- 11005722 TI - Stereological estimation of volume-weighted mean glomerular volume from arbitrary sections of the equine kidney. AB - Mean glomerular volume has previously been estimated, using stereological techniques, specifically the point-sampled intercept (PSI), either from isotropic or from vertical sections. As glomeruli are approximately spherical structures, the same stereological technique was carried out on vertical and arbitrary sections to determine whether section orientation had any effect on mean glomerular volume estimation. Equine kidneys from 10 individuals were analysed using the PSI method of estimating volume-weighted mean glomerular volume (MGV); for each kidney, arbitrary and vertical sections were analysed. MGVs were not significantly different between arbitrary and vertical sections (P = 0.691) when analysing the data with the paired t test; when plotting MGV estimates from arbitrary sections against those from vertical sections the intercept was found not to be significantly different from zero (P > 0.8) and the slope of the regression line not to be significantly different from 1.0 (P > 0.4). For the estimation of MGV in equine kidneys using PSI, arbitrary sections may be used if it is not possible to use isotropic or vertical sections, but some caution must be exercised in the interpretation of results so gained. PMID- 11005721 TI - Macrophages and apoptosis in the stellate reticulum of the rat enamel organ. AB - Tooth germs of upper first molars of 1, 3, and 5-d-old rats, fixed in formaldehyde, were stained for the detection of apoptosis by the TUNEL method, and by the azo-dye method for the demonstration of acid phosphatase. For conventional light and electron microscopy the specimens were fixed in glutaraldehyde-formaldehyde and embedded in glycol methacrylate and Araldite. Results showed that macrophages, present in the stellate reticulum, contained basophilic bodies and TUNEL-positive globules, i.e. apoptotic bodies, in their interior. Macrophages also possessed strong acid phosphatase activity. Electron microscopy showed the presence of large vacuoles inside the macrophages containing dense fragmented material. Taken together these results suggest that the intra-epithelial macrophages of the stellate reticulum engulf apoptotic bodies. PMID- 11005723 TI - Variation in shape of the lingula in the adult human mandible. AB - The lingulae of both sides of 165 dry adult human mandibles, 131 males and 34 females of Indian origin, were classified by their shape into 4 types: 1, triangular; 2, truncated; 3, nodular; and 4, assimilated. Triangular lingulae were found in 226 (68.5%) sides, truncated in 52 (15.8%), nodular in 36 (10.9%) and assimilated in 16 (4.8%) sides. Triangular lingulae were found bilaterally in 110, truncated in 23, nodular in 17 and assimilated in 7 mandibles. Of the remaining 8 mandibles with different appearances on the 2 sides, 6 had a combination of triangular and truncated and 2 had nodular and assimilated. The incidence of triangular and assimilated types in the male and female mandibles are almost equal. In the truncated type it was double in the male mandibles while the nodular type was a little less than double in the female mandibles. PMID- 11005724 TI - The world of oral research: how do we improve it? PMID- 11005725 TI - Necessary correction of bacterial names? PMID- 11005726 TI - Biodegradation of Bis-GMA. PMID- 11005727 TI - A Philadelphia story--featuring Ned Williams: Microbiology at the University of Pennsylvania School of Dental Medicine. PMID- 11005728 TI - Interfacial chemistry of the dentin/adhesive bond. AB - To date, the dentin/adhesive (d/a) bond has primarily been studied by morphologic analysis in conjunction with bond strength measurement. Although these analyses have enhanced our understanding, numerous questions about the chemistry have not been answered. The purpose of this study was to determine, at the molecular level, quantitative differences in the composition of the d/a interface formed under "wet" bonding conditions. The occlusal one-third of the crown was removed from 10 extracted, unerupted human third molars. The prepared dentin surfaces were treated, per manufacturers' instructions, with either Single Bond (3M) or One-Step adhesive (Bisco). Three-micron-thick sections of the d/a interface were cut and stained with Goldner trichrome for light microscopy. Companion slabs were analyzed with micro-Raman spectroscopy; the sample was placed at the focus of a 100x microscope objective, and spectra were acquired at 1-microm intervals across the d/a interface. Reference spectra were collected on model compounds of type I collagen and adhesive; the relative ratios of the integrated intensities of spectral features from adhesive and collagen were determined and plotted as a function of wt% adhesive. The same ratios were determined for the interface samples; by comparing these ratios with the calibration curve generated from the model compounds, we determined the percent of adhesive as a function of spatial position across the d/a interface. The relative percent of Single Bond adhesive was < 50% throughout more than half of the hybrid layer; One Step adhesive was > or = 50% throughout most of the hybrid. The results from this study provide the first direct chemical evidence of phase separation in a dentin adhesive and its detrimental effect on the dentin/adhesive bond. PMID- 11005729 TI - Temperature dependence of the electrical resistance of sound and carious teeth. AB - Temperature variations are expected to influence measurement error in electrical resistance of teeth. It was the aim of this study to determine the changes in electrical behavior of extracted human teeth due to temperature changes in the range of room temperature to intra-oral temperature. Nine extracted teeth were selected, and the occlusal or an approximal surface was chosen for measurement. Carious involvement of the surfaces ranged from sound to cavitated. Electrical impedance spectroscopy sweeps in a frequency range of about 100 kHz to 10 Hz were completed at selected temperatures between 22 degrees C and 40 degrees C. After fitting the data to equivalent circuits that yielded parameter values for components of the equivalent circuit, we calculated the dc bulk resistance (Rh). The temperature dependence of Rb of the surfaces with different carious involvement was very similar, and the mean drop of Rb from 20 to 35 degrees C was 45% (SD 2%). It was concluded that the electrical resistance of sound and carious tooth surfaces is inversely related to temperature. PMID- 11005730 TI - Clinical, radiographic, and genetic evaluation of a novel form of autosomal dominant oligodontia. AB - A frameshift mutation recently identified within the paired domain of the transcription factor, PAX9, has been linked to a unique form of oligodontia in a single, multigenerational family (Stockton et al., 2000). We now describe the phenotypic and segregation analyses of this remarkable kindred, the initial approach taken to identify a candidate gene involved in this form of oligodontia, and the power of this single-family pedigree to generate significant linkage in a genome search. Of the 43 family members enrolled in this study, 21 individuals were affected with several congenitally missing permanent teeth. The pattern of inheritance of the oligodontia trait suggested the involvement of a single gene bearing a dominant mutation. To various degrees, affected members lacked permanent first, second, and third molars in all four quadrants. Several individuals with missing molars also lacked second premolars- most commonly, maxillary second premolars and mandibular central incisors. To the best of our knowledge, this pattern of non-syndromic, familial tooth agenesis has not been previously described in the literature. Since a missense mutation in the homeobox gene, MSX1, was previously linked to tooth agenesis in a single family lacking second premolars and third molars, we performed a mutational analysis of MSX1 by PCR. The absence of a mutation in exons 1 and 2 of MSX1 suggested that allelic mutations in the coding region of MSX1 are not associated with this phenotypically distinct form of oligodontia. Computer simulation of linkage analysis further proved that this pedigree alone was sufficient to generate a significant result for a total genome scan. PMID- 11005731 TI - Unique enamel phenotype associated with amelogenin gene (AMELX) codon 41 point mutation. AB - Different mutations in the amelogenin gene (AMELX) result in the markedly different enamel phenotypes that are collectively known as amelogenesis imperfecta (AI). We hypothesize that unique phenotypes result from specific genetic mutations. The purpose of this study was to characterize the enamel compositional and structural features associated with a specific AMELX mutation in three families with X-linked AI. We performed mutational analysis by amplifying AMELX exons and sequencing the products. Permanent and primary affected (N = 6) and normal (N = 3) teeth were collected and examined by light, scanning, and transmission electron microscopy. Enamel proteins were evaluated by immunolocalization of amelogenin and amino acid analysis. AI-affected individuals all shared a common AMELX point mutation (C to A change at codon 41). The dental phenotypic findings were remarkably consistent in all affected individuals. The AI enamel was opaque, with numerous prism defects or holes encompassing the entire prism width. Affected crystallites appeared more radiolucent and morphologically less uniform, compared with that of normal enamel. Immunogold labeling with anti-amelogenin antibodies localized amelogenin to the crystallites but not to the inter-crystalline spaces. No immunogold labeling was seen in normal enamel. There was an increased and amelogenin-like protein content in AI enamel (0.95%) compared with normal enamel (0.13%). We conclude that this codon 41 C to A missense point mutation, in a highly conserved region of the AMELX gene, results in a remarkably consistent phenotype. PMID- 11005732 TI - Cementum attachment protein enriches putative cementoblastic populations on root surfaces in vitro. AB - We tested the capacity of cementum attachment protein (CAP) to recruit putative cementoblastic populations to root surfaces in vitro by determining the phenotypic expression of periodontal ligament cloned cell populations. The clones were derived from cells that attached to either CAP-coated (experimental) or uncoated (control) root slices. Root slices were co-cultured with primary human periodontal ligament cells. Cloned and parent populations were analyzed for their capacity to express alkaline phosphatase (AP), osteopontin, bone sialoprotein (BSP), and CAP and to form mineralized tissue in vitro. The percentage of CAP- and BSP-positive clones was significantly higher in the experimental clones than in the controls. The percentage of cells positive for AP, BSP, and CAP was higher in the experimental clones than in their control counterparts. Mineralized tissue formation was observed only in the cell populations derived from the CAP-coated root slices. These results indicate that CAP is capable of recruiting putative cementoblastic populations on root slices in vitro and therefore might play an important role in cementogenesis during periodontal homeostasis and wound healing. PMID- 11005733 TI - Reactive oxygen species participation in experimentally induced arthritis of the temporomandibular joint in rats. AB - In the temporomandibular joint (TMJ), it has been hypothesized that mechanical stresses lead to the oxidative stress of articular tissues. It has also been postulated that cells pertinent to arthritis-including endothelial cells and synovial cells-when stimulated by mechanical stresses and/or pro-inflammatory cytokines, promote oxidative damage. To determine the involvement of reactive oxygen species (ROS) in the diseased joint, we studied the generation of ROS in synovial fluid (SF) from interleukin-1alpha (IL-1alpha)-induced TMJ arthritis by electron spin resonance (ESR) spectroscopy, using the spin trap 5,5-dimethyl-1 pyrroline N-oxide (DMPO). The TMJ arthritis was experimentally induced in rats by the injection of human recombinant IL-1alpha into the TMJ; control rats were treated with normal saline solution. We found that the detected radicals in the collected SF were identified as a 1:2:2:1 quartet, characteristic of the hydroxyl radical-DMPO spin adduct. The ESR signal intensity of the hydroxyl radical-DMPO spin adduct in the SF from IL-1-treated rats was significantly higher than that from the control rats (P < 0.01). The results of ESR study also showed that hydroxyl radical (HO*) was increased in a time-dependent fashion in the presence of superoxide anion radical (O2*-) scavenger superoxide dismutase (SOD); the formation of DMPO-HO* was strongly inhibited by the iron chelater deferoxamine. We could measure higher levels of free iron (Fe2- and Fe3-) in the SF from TMJ arthritis than in that from controls (P < 0.05). Analysis of the data obtained from the present study suggests that the HO* radical detected in SF from IL-1 induced TMJ arthritis is generated via a modified Haber-Weiss reaction (biological Fenton reaction) in which O2*- can subsequently result in the production of H2O2 through dismutation reaction by SOD. Thus, HO* may be generated from the reaction of resultant H2O2 with free iron ions. The results presented here provide the first evidence of involvement of ROS in IL-1-induced TMJ arthritis. PMID- 11005734 TI - Proviral HIV-1 DNA in gingival crevicular fluid of HIV-1-infected patients in various stages of HIV disease. AB - The oral cavity is rarely reported to be a site of human immunodeficiency virus (HIV) transmission, despite detectable virus in saliva and relatively frequent prevalence of periodontal disease in HIV-infected persons yielding increased excretion of mononuclear-cell-enriched gingival fluid. To search for possible sources of HIV in saliva, and using the polymerase chain-reaction technique, we sought the presence and shedding patterns of proviral HIV-1 DNA in gingival crevicular fluid in a group of patients previously determined as HIV-1 seropositive. Periodontal status at the collection sites was monitored by several clinical parameters, including Plaque Index, Gingival Index, probing depth, and clinical attachment loss. Gingival crevicular fluid samples were collected by means of paper points. Proviral HIV-1 DNA was detected in the gingival fluid of 17 out of 35 HIV-1-infected patients. Its detection correlated significantly with higher plasma HIV-1 RNA viral load (p = 0.03) and not with peripheral blood CD4+ cell count, the presence of blood in gingival fluid, or oral lesions. There was a significant correlation between clinical attachment loss at the sites of fluid collection and plasma HIV-1 RNA viral load (p = 0.002), and borderline correlation between the latter and probing depth (p = 0.54) in the group of patients harboring proviral HIV-1 DNA in gingival crevicular fluid. The results of our study suggest that mononuclear cells present in gingival crevicular fluid and harboring proviral HIV-1 DNA could represent a potential source of HIV-1 in the presence or absence of local bleeding, especially in persons with advanced HIV infection and increased loss of clinical attachment. PMID- 11005735 TI - The prevalence of xerostomia and salivary gland hypofunction in a cohort of HIV positive and at-risk women. AB - The association of xerostomia and salivary gland hypofunction with HIV infection has been established for men but not for women. We investigated the prevalence of these conditions in a national cohort (n = 733) of HIV-positive and at-risk HIV negative women. Participants in this prospective cross-sectional study were recruited from the Women's Interagency HIV Study (WIHS) at five outpatient USA clinics. Xerostomia was assessed based on "yes" responses to a dry-mouth questionnaire. Samples of unstimulated whole and chewing-stimulated whole saliva were collected under standardized conditions. The major salivary glands were also evaluated clinically. The prevalence of dry-mouth complaint, the absence of saliva upon palpation, and zero unstimulated whole saliva (flow rate = 0 mL/min) were significantly (p = 0.001) higher in HIV-positive women. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher in HIV positive women vs. HIV-negative women (OR = 2.86; 95% CI, 1.23 to 6.63). Significant (p = 0.03) univariate association was found between zero unstimulated whole saliva and CD4 counts. Adjusted odds of zero unstimulated whole saliva were significantly (p = 0.02) higher for HIV-positive women with CD4 < 200 compared with those with CD4 > 500 (OR = 2.61; 95% CI, 1.17 to 5.85). Chewing-stimulated flow rates were not significantly different between seropositive and seronegative women. The prevalence of xerostomia and salivary gland hypofunction appears to be significantly higher in HIV-positive women relative to a comparable group of at risk seronegative women. Immunosuppression levels measured by CD4 cell counts are significantly associated with xerostomia and salivary gland hypofunction in a population of HIV-positive women. PMID- 11005736 TI - Intra-oral distribution of dental fluorosis in Newburgh and Kingston, New York. AB - Previous studies of intra-oral distribution of dental fluorosis in low-fluoride areas reported that teeth that formed later in life were more frequently affected compared with the early-forming teeth. The steady increase of plasma fluoride with age, even under constant fluoride exposure, has been suggested as a possible mechanism for this clinical manifestation. To determine the intra-oral distribution of Dean's Index scores and the effect of fluoride exposure on early- and late-forming teeth, we analyzed data collected on 2193 seven- to 14-year-old lifelong residents of fluoridated or non-fluoridated areas. Logistic regression procedures were used to determine the effects of fluoridation, early brushing, daily supplements, and other socio-demographic variables on early- and late forming teeth. The results show that the occurrence of very mild or greater levels of fluorosis in the upper anterior teeth was 7 to 10% in the fluoridated area and 5 to 9% in the nonfluoridated area. In the fluoridated area, the occurrence of fluorosis increased from anterior to posterior teeth. Both early- and late-forming teeth were affected by exposure to fluoridation, daily fluoride supplement use, or brushing before the age of two years. This analysis showed that the esthetic consequence of exposure to multiple sources of fluoride was less dramatic, as evidenced by the lower frequency in upper anterior teeth compared with posterior teeth. The longer maturation process of the posterior teeth and the thicker enamel appear to be the likely explanation for the higher occurrence of dental fluorosis in posterior teeth. PMID- 11005737 TI - Cortical potentials associated with voluntary mandibular movements. AB - Movement-related cortical potentials (MRCPs) are negative potentials over the scalp, which gradually increase prior to voluntary movements, and might be applied to elucidate the cortical efferent function of the mandibular movements. We compared the MRCPs accompanying various mandibular movements to study the motor control mechanism underlying these movements. Electroencephalograms (EEGs) were recorded from 11 electrodes placed over the scalp (F3, Fz, F4, T3, C3, Cz, C4, T4, P3, Pz, and P4), according to the International 10-20 System, and electromyograms (EMGs) were obtained from surface electrodes over the masseter muscle and the anterior belly of the digastric muscle. Ten healthy subjects were requested to make brisk and self-paced mandibular movements in 4 different directions (mouth-opening and -closing, and left and right lateral movements). We obtained MRCPs by averaging the EEG, using the visually determined EMG onset as a trigger signal. In all the movements, a slowly increasing, bilaterally widespread negativity starting 1.5 to 2.0 sec before the EMG onset (Bereitschaftspotential, or BP proper) was observed, with the maximum over the vertex region. The negative slope (NS') occurred about 300 to 700 msec before the EMG onset. The cortical maps of BP/NS' (BP and NS' combined), immediately prior to the mouth-opening and closing, showed a symmetrical distribution, whereas that for the lateral movements showed a tendency of predominance over the hemisphere ipsilateral to the direction of the movement. BP/NS' amplitudes at the onset of movement differed significantly or tended to do so between open, close, and lateral movements, suggesting that MRCP recordings may thus provide a means to explore the role of the cerebral cortex in the control of mandibular movements. PMID- 11005738 TI - Biting and chewing in overdentures, full dentures, and natural dentitions. AB - It has been suggested that the provision of dental implants can improve the oral function of subjects with severely resorbed mandibles, possibly restoring function to the level experienced by satisfied wearers of conventional complete dentures. Nevertheless, a quantitative comparison has never been made and can be drawn from the literature only with difficulty, since studies differ greatly in methodology. To make such a comparison, we measured bite force and chewing efficiency by using identical methods in subjects with overdentures, complete full dentures, and natural dentitions. Our results indicated that bite forces achieved with overdentures on dental implants were between those achieved with artificial and natural dentitions. Chewing efficiency was significantly greater than that of subjects with full dentures (low mandible), but was still lower than that of subjects with full dentures (high mandible) and overdentures on bare roots. Differences in the height of the mandible revealed significant differences in chewing efficiency between the two full-denture groups. Furthermore, subjects with a shortened dental arch exerted bite forces similar to those of subjects with a complete-natural dentition, but their chewing efficiency was limited due to the reduced occlusal area. For all groups combined, a significant correlation was found between maximum bite force and chewing efficiency. Nearly half of the variation in chewing efficiency was explained by bite force alone. PMID- 11005739 TI - Effect of food size on the movement of the mandibular first molars and condyles during deliberate unilateral mastication in humans. AB - To date, the effect of food size on the movement of the mandibular first molars and condyles during chewing has not been fully examined due to methodological problems. The purpose of the present study was to examine the previously unknown effect of food size on masticatory jaw movement. Using a face bow, light-emitting diodes, and optical cameras, we recorded, in 16 young adults with good occlusion, mandibular movement for the first 10 strokes during the unilateral chewing of similarly shaped hard gummy jellies weighing 5 g and 10 g, respectively. The chewing cycle time for the 10-g jelly was significantly longer than that for the 5-g jelly. The jaw-closing and -opening maximum velocities, gapes at the maximum velocities, and maximum gape were significantly faster and larger when 10-g gummy jellies were chewed, compared with results with 5-g jellies, at the mandibular first molar on the chewing side and the condyle on the non-chewing side. With the exception of the velocity, similar tendencies were observed at the molar on the non-chewing side. However, such significant differences were not detected at the condyle on the chewing side. The mandibular first molar on the chewing side was that most affected by food size, and the mean value of the maximum gape coincided approximately with the height of each jelly. These results suggest that humans chew hard coherent food such that the mandibular teeth that come into contact with the food open to a height equivalent to that of the food bolus, and that the changes in movement of the other parts of the mandible are minimized, ensuring efficient mastication. PMID- 11005740 TI - Use of radiology practice guidelines and compliance with accreditation standards in US and Canadian dental schools. AB - In 1992 and 1997, all US and Canadian dental schools were surveyed by mail regarding the preferred initial radiographic examination prescribed for non emergency, comprehensive-care patients (dentulous adults, edentulous adults, and children). In both survey years, a minority of US and Canadian dental schools reported using selection criteria for dentulous adults and children, while nearly all schools reported doing so for edentulous adults. The purpose of this secondary analysis was to investigate the relationship between the use of radiology selection criteria (vs. predetermined routine examinations) in US and Canadian dental school clinics and three factors: (1) the credentials of the chief-of-service, (2) institutional funding, and (3) geographic region. "Credentials of the chief-of-service" is the single factor significantly related to the distributions of radiographic examinations prescribed for dentulous adults in both years (Fisher exact test, p < or = 0.02). There are no statistically significant relationships for edentulous adults or children in either year. Multivariate analyses (logistic regression) of the 1997 data reveal that institutions with a credentialed chief-of-service are 2.39 times more likely to report using selection criteria than institutions with a noncredentialed chief-of service; private institutions are 1.13 times more likely than public institutions, and Canadian schools are 3.65 times more likely than US schools. A similar trend was identified for children. Analysis of the 1992 data revealed similar trends for the credentials of the chief-of-service and the geographic region, but showed no association between institutional funding source and the use of selection criteria. Contrary to accreditation standards, most US and Canadian dental schools obtained pre-determined routine radiographic examinations on most new patients. However, the presence of a credentialed chief-of-service had a positive effect on the use of selection criteria for dentulous adults and children. PMID- 11005741 TI - "DentalResearch2000.org". PMID- 11005742 TI - The Leeds Assessment Scale of Handicap: its operationalisation, reliability, validity and responsiveness in in-patient rehabilitation. AB - PURPOSE: This paper describes the application of the handicap dimension of the International Classification of Impairments, Disabilities and Handicaps. METHOD: A method of assessing four of the six roles (or areas of expected competency) was developed and applied to an in-patient population, aged 16 to 65 years, receiving specialist rehabilitation following neurological damage. Handicap assessment was a collaborative process by a multidisciplinary team. Difficulties were experienced in measuring inter-rate reliability in the busy, clinical setting. RESULTS: We conclude that handicap assessment in an in-patient rehabilitation unit which uses a multidisciplinary approach to treating patients with complex physical, sensory and cognitive problems is both feasible and useful. CONCLUSION: The developed scale, The Leeds Assessment Scale of Handicap, provides a valid and valuable summary of human disadvantage, complementing the common disability assessments. PMID- 11005743 TI - Coping with illness after brain diseases--a comparison between patients with malignant brain tumors, stroke, Parkinson's disease and traumatic brain injury. AB - PURPOSE: This study investigates coping styles in patients suffering from different brain disorders (malignant brain tumors, stroke, Parkinson's disease and traumatic brain injury). METHOD: In a combined analysis of four prospective studies we investigated 21 patients with malignant glioma at the end radiochemotherapy, 30 patients one year after ischemic stroke, 54 patients suffering from various stages of Parkinson's disease, and 58 patients 6 to 8.5 months after traumatic brain injury. The assessment of coping with illness was based on the Freiburg Questionnaire on Coping with illness. RESULTS: With few exceptions, coping styles did not differ across the various brain pathologies. Differences occurred with respect to 'active, problem-oriented coping' (decreased in stroke patients), and coping by search for religious relief or quest for sense (increased in patients with Parkinson's disease). CONCLUSION: Differences in coping styles could be mainly related to age and social factors. Individual coping strategies seem only to be little related to the type of brain pathology. PMID- 11005744 TI - Rehabilitation of stroke patients with apraxia: the role of additional cognitive and motor impairments. AB - PURPOSE: The present study investigated which additional cognitive and motor impairments were present in stroke patients with apraxia and which of these factors influenced the effects of treatment. METHOD: A group of 33 patients with apraxia were treated according to the guidelines of a therapy programme based on teaching patients strategies to compensate for the presence of apraxia. Patients were treated at occupational therapy departments in general hospitals, rehabilitation centres and nursing homes. The outcome of the strategy training was studied in a pre-post test design; measurements were conducted at baseline and after 12 weeks of therapy. The pretreatment scores of the patients with apraxia were compared to normscores and scores of a control group of patients without apraxia (n = 36) to investigate which impairments are present. The following variables were analysed in order to determine which factors influence outcome: additional neuropsychological deficits (comprehension of language, cognitive impairments due to dementia, neglect and short term memory), level of motor functioning, severity of apraxia and performance on activities of daily living (ADL), and some relevant patient characteristics (gender, age, type of stroke, time since stroke, and location of treatment). RESULTS: The results showed that the presence of apraxia is associated with the presence of additional cognitive and motor impairments. The successful outcome of strategy training was not negatively influenced by cognitive comorbidity. The outcome seemed to be more prominent in patients who were more severely impaired at the start of rehabilitation in terms of the degree of motor impairments, the severity of apraxia and the initial ADL dependence. The ADL observations, however, displayed a ceiling effect, which was taken into account in discussing the results. Demographic variables, especially age, did not predict the outcome of treatment. CONCLUSIONS: We suggest that the effect of this training is stronger in more severely disabled patients. However, neither the presence of additional cognitive impairments nor the severity of motor problems nor old age should be an indication for refraining from treating apraxia. PMID- 11005745 TI - Spiritual issues associated with traumatic-onset disability. AB - PURPOSE: The present study used a qualitative approach to discover the concepts used by people with a sudden-onset disability to express issues of spirituality. Using a cross-sectional time frame, the study sought to understand how spiritual issues were experienced at the onset of a disability from the perspective of the disabled person him or herself. The study also compared the perceptions of brain injury respondents to spinal cord injury respondents. METHOD: The qualitative approach was chosen to capture a detailed understanding of perceptions about spirituality, and the relationship of the disability to the disabled person's spiritual beliefs. The sample consisted of 16 individuals, all of whom had experienced a spinal cord injury or a brain injury. RESULTS: The main findings of the study were as follows: (1) Participants recovering from a disability described spiritual issues relating to five themes which arose directly from the data (awareness, closeness, trust, purpose, vulnerability) and relating to three relationships found throughout the literature on spirituality (intrapersonal, interpersonal and transpersonal). These eight dimensions can be combined to form a matrix, which serves as a framework for considering spiritual issues associated with disability; and (2) There were differences in the spiritual concepts described by the participants with brain injury compared to those with spinal cord injury. In particular, those with brain injuries seemed to place greater emphasis on the importance of their families, and to be more aware of the need for trust in view of their memory deficits. CONCLUSIONS: These results represent one of the earliest attempts to take an empirical approach to the development of theory in the area of spirituality and disability. Building on theory developed in other populations (palliative care and ageing), the study offers a theoretical model for clinicians, educators and researchers to better understand spirituality in the context of disability. PMID- 11005746 TI - Response of prolonged flaccid paralysis to FNS rehabilitation techniques. AB - PURPOSE: The purpose of this study was to investigate the response of muscles with prolonged flaccid paralysis (a year after stroke) to two types of treatment: (1) functional neuromuscular stimulation (FNS) with surface electrodes; and (2) FNS with intramuscular (IM) electrodes (FNS-IM). A second purpose was to compare FNS-gait versus volitional gait (no FNS activation). METHOD: We used a single case study design; our patient was age 72, with flaccid paralysis of knee flexors and ankle dorsiflexors. RESULTS: Following four months of treatment with surface stimulation, there was no change in muscle function or gait. Following treatment with FNS-IM, the patient regained partial volitional control of knee flexors and dorsiflexors; untreated muscles did not change. CONCLUSION: FNS-gait provided more normal knee and ankle dorsiflexion during swing phase versus volitional gait swing phase (no FNS activation). PMID- 11005747 TI - Severe anaemia: implications for functional recovery during rehabilitation. AB - PURPOSE: This case report examines the hospital course and functional recovery of a geriatric patient with severe anaemia undergoing rehabilitation following total hip replacement. It serves to highlight the need for further study of the impact of anaemia on recovery in the rehabilitation setting as well as the importance of developing clinical guidelines for the appropriate medical management of anaemia in this patient population. METHODS: Single case report. RESULTS: The course of recovery during rehabilitation in a geriatric patient with severe post-operative anaemia was notably different from other geriatric patients undergoing rehabilitation following total hip replacement. The patient demonstrated a markedly reduced tolerance for therapies and prolonged length of stay, but did not experience cardiopulmonary complications during intensive rehabilitation. Her hematocrit responded nicely to treatment with human recombinant erythropoietin. Despite a prolonged recovery period, the patient ultimately progressed well and achieved a good functional outcome. CONCLUSIONS: The impact of anaemia on functional recovery in the acute inpatient rehabilitation setting as well as the theoretical risk of increased morbidity and mortality during prescribed therapeutic exercise has not been closely examined in the literature. Further study is indicated to examine the implications for anaemia on functional recovery and cardiopulmonary complications during rehabilitation. General guidelines should be developed for the management of anaemia in the rehabilitation setting. PMID- 11005748 TI - [Smoking in childhood and adolescence. Role of the pediatrician in control and prevention]. PMID- 11005749 TI - [Pediatric cardiopulmonary resuscitation equipment in the resuscitation cart or trolley. Spanish Group of Pediatric and Neonatal Cardiopulmonary Resuscitation]. PMID- 11005750 TI - Fibrogenesis. IV. Fibrosis and inflammatory bowel disease: cellular mediators and animal models. AB - The cellular mediators of intestinal fibrosis and the relationship between fibrosis and normal repair are not understood. Identification of the types of intestinal mesenchymal cells that produce collagen during normal healing and fibrosis is vital for elucidating the answers to these questions. Acute injury may cause normal mesenchymal cells to convert to a fibrogenic phenotype that is not maintained during normal healing but may lead to fibrosis when inappropriately sustained. Proliferation of normal or fibrogenic mesenchymal cells may lead to muscularis overgrowth associated with fibrosis. The presence of increased numbers of vimentin-positive cells within fibrotic, hypertrophied muscularis in Crohn's disease suggests that changes in mesenchymal cell phenotype and number may indeed be associated with fibrosis. Fibrosis is induced in rats by peptidoglycan polysaccharides or trinitrobenzene sulfonic acid-ethanol administration, but inducing fibrosis in mice has been technically challenging. The development of current mouse models of colitis, such as dextran sodium sulfate or trinitrobenzene sulfonic acid-ethanol administration, into models of fibrosis will allow us to use genetic manipulation to study molecular mediators of fibrosis. PMID- 11005751 TI - Molecular physiology and pathophysiology of tight junctions III. Tight junction regulation by intracellular messengers: differences in response within and between epithelia. AB - Tight junction permeability differs with the type of permeants, their size, and their charge. Selective changes in permeability do occur, and they illustrate the diversity in functional reactions of tight junctions. This suggests that special structures in the tight junctions are involved. More and more structural components of the tight junctions are becoming known. The divergence in behavior of native tissue and filter-grown epithelial monolayers with respect to the effects of intracellular messengers offers the possibility to relate structure and function. In addition to the tools for conventional permeability studies, probes have become available to detect changes in activation of intracellular effector proteins such as the protein kinase C isotypes, and with in situ imaging techniques the way is open for a functional approach in the study of tight junctions. PMID- 11005752 TI - Cerulein upregulates ICAM-1 in pancreatic acinar cells, which mediates neutrophil adhesion to these cells. AB - Neutrophil infiltration into the pancreas is a key event in pancreatitis. Here we show that intercellular adhesion molecule-1 (ICAM-1), which regulates neutrophil adhesion, is present on rat pancreatic acinar cells, is upregulated by a hormone (cerulein) and mediates direct binding of neutrophils to acinar cells. ICAM-1 was upregulated in pancreas of rats with experimental pancreatitis induced by supramaximal doses of cerulein. Furthermore, cerulein time and dose dependently stimulated expression of ICAM-1 mRNA and protein in isolated pancreatic acinar cells. Inhibitory analysis showed that activation of transcription factor nuclear factor-kappaB (NF-kappaB) was involved in ICAM-1 upregulation by cerulein, but NF kappaB did not mediate basal expression of ICAM-1 mRNA in acinar cells. With an adhesion assay, we found that neutrophils bind to isolated pancreatic acinar cells and that cerulein upregulates the extent of adhesion. Neutralizing ICAM-1 antibody blocked neutrophil binding to both control and cerulein-stimulated acinar cells, suggesting ICAM-1 involvement in this adhesion. Thus the acinar cell is capable of targeting neutrophils to its surface, a process that may be important for inflammatory and cell death responses in pancreatitis and other pancreatic disorders. PMID- 11005753 TI - Effects of gamma-aminobutyric acid on secretagogue-induced exocrine secretion of isolated, perfused rat pancreas. AB - Because GABA and its related enzymes have been determined in beta-cells of pancreas islets, effects of GABA on pancreatic exocrine secretion were investigated in the isolated, perfused rat pancreas. GABA, given intra-arterially at concentrations of 3, 10, 30, and 100 microM, did not exert any influence on spontaneous or secretin (12 pM)-induced pancreatic exocrine secretion. However, GABA further elevated CCK (10 pM)-, gastrin-releasing peptide (100 pM)-, or electrical field stimulation-induced pancreatic secretions of fluid and amylase dose dependently. The GABA (30 microM)-enhanced CCK-induced pancreatic secretions were completely blocked by bicuculline (10 microM), a GABA(A) receptor antagonist, but were not affected by saclofen (10 microM), a GABA(B) receptor antagonist. The enhancing effects of GABA (30 microM) on CCK-induced pancreatic secretions were not changed by tetrodotoxin (1 microM) but were partially reduced by cyclo-(7-aminoheptanonyl-Phe-D-Trp-Lys-Thr[BZL]) (10 nM), a somatostatin antagonist. In conclusion, GABA enhances pancreatic exocrine secretion induced by secretagogues, which predominantly induce enzyme secretion, via GABA(A) receptors in the rat pancreas. The enhancing effect of GABA is partially mediated by inhibition of islet somatostatin release. PMID- 11005754 TI - Endothelin-mediated vasoconstriction in postischemic newborn intestine. AB - We previously suggested that the profound, sustained vasoconstriction noted in 3 day-old swine intestine after a moderate episode of ischemia-reperfusion (I/R) reflects the unmasking of underlying constrictor tone consequent to a loss of endothelium-derived nitric oxide (NO). In this study, we sought to determine whether endothelin-1 (ET-1) was the unmasked constrictor and whether selective loss of endothelial ET(B) receptors, which mediate NO-based vasodilation, participated in the hemodynamic consequences of I/R in newborn intestine. Studies were performed in innervated, autoperfused intestinal loops in 3- and 35-day-old swine. Selective blockade of ET(A) receptors with BQ-610 had no effect on hemodynamics under control conditions; however, when administered before and during I/R, BQ-610 significantly attenuated the post-I/R vasoconstriction and reduction in arteriovenous O(2) difference in the younger group. In 3-day-old intestine, reduction of intestinal O(2) uptake to a level similar to that noted after I/R by lowering tissue temperature had no effect on the response to BQ-610 or ET-1, indicating that the change in response to BQ-610 noted after I/R was not simply consequent to the reduction in tissue O(2) demand. In studies in mesenteric artery rings suspended in myographs, we observed a leftward shift in the dose-response curve for ET-1 after selective blockade of ET(B) receptors with BQ-788 in 3- but not 35-day-old swine. Rings exposed to I/R in vivo behaved in a manner similar to control rings treated with BQ-788 or endothelium-denuded non I/R rings. PMID- 11005755 TI - Rectal sensitivity assessed by a reflexologic technique: further evidence for two types of mechanoreceptors. AB - We previously showed that slow-ramp rectal distensions induce graded inhibitions of the somatic nociceptive RIII reflex recorded from the lower limb, which correlated with both distension volume and visceral sensation. In contrast, rapid phasic rectal distensions induced facilitatory or biphasic effects (i.e., facilitations followed by inhibitions) depending on the level of distension. To examine the role of mucosal and serosal rectal mechanoreceptors in these viscerosomatic interactions, we analyzed, in six healthy volunteers, the effects of both types of rectal distension on the RIII reflex after topical application of lidocaine or placebo administered in a double-blind and crossover fashion. Inhibitions of the RIII reflex induced by both slow-ramp and rapid distensions were strongly reduced after administration of lidocaine but not after placebo. In contrast, facilitations of the RIII reflex observed during the initial phase of rapid distensions were not modified after lidocaine or placebo applications. These results suggest that inhibitions, but not facilitations, of the nociceptive RIII reflex triggered by rectal distensions depend preferentially on the activation of superficial mucosal receptors. This reflexologic technique might thus represent an interesting tool for studying the role of the different rectal mechanoreceptors involved in visceral sensations. PMID- 11005756 TI - GH decreases hepatic amino acid degradation after small bowel resection in rats without enhancing bowel adaptation. AB - Growth hormone (GH) treatment in short bowel syndrome is controversial, and the mechanisms of a possible positive effect remain to be elucidated. Rats were randomly subjected to either an 80% jejunoileal resection or sham operation and were given either placebo (NaCl) or biosynthetic rat GH (brGH). The in vivo capacity of urea nitrogen synthesis (CUNS) and the expression of urea cycle enzymes were measured and related to changes in body weight and adaptive growth in ileal segments on days 7 and 14. Ileal segments were examined by unbiased stereological techniques. brGH treatment decreased CUNS among the resected rats by 19% (P<0.05) and 36% (P<0.05) on days 7 and 14, respectively. The mRNA levels of urea cycle enzyme genes were not influenced by brGH treatment. brGH treatment did not increase the adaptive growth in the ileal segments. In conclusion, we found that GH treatment decreased the accelerated postoperative hepatic amino acid degradation in experimental short bowel syndrome without enhancing the morphological intestinal adaptation. PMID- 11005757 TI - Intestinal ion transport in NKCC1-deficient mice. AB - The Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) located on the basolateral membrane of intestinal epithelia has been postulated to be the major basolateral Cl(-) entry pathway. With targeted mutagenesis, mice deficient in the NKCC1 protein were generated. The basal short-circuit current did not differ between normal and NKCC1 -/- jejuna. In the -/- jejuna, the forskolin response (22 microA/cm(2); bumetanide insensitive) was significantly attenuated compared with the bumetanide sensitive response (52 microA/cm(2)) in normal tissue. Ion-replacement studies demonstrated that the forskolin response in the NKCC1 -/- jejuna was HCO(3)(-) dependent, whereas in the normal jejuna it was independent of the HCO(3)(-) concentration in the buffer. NKCC1 -/- ceca exhibited a forskolin response that did not differ significantly from that of normal ceca, but unlike that of normal ceca, was bumetanide insensitive. Ion-substitution studies suggested that basolateral HCO(3)(-) as well as Cl(-) entry (via non-NKCC1) paths played a role in the NKCC1 -/- secretory response. In contrast to cystic fibrosis mice, which lack both basal and stimulated Cl(-) secretion and exhibit severe intestinal pathology, the absence of intestinal pathology in NKCC1 -/- mice likely reflects the ability of the intestine to secrete HCO(3)(-) and Cl(-) by basolateral entry mechanisms independent of NKCC1. PMID- 11005758 TI - Effect of galanin and galanin antagonists on peristalsis in esophageal smooth muscle in the opossum. AB - Galanin, a neuropeptide that is widely distributed in the esophageal nerves, is known to exert a neuromodulatory action in the gut. These studies examined the effect of galanin and galanin antagonists on esophageal peristalsis in anesthetized opossums in vivo. Intraluminal esophageal pressures were recorded at 1, 3, 5, 7, and 9 cm above the lower esophageal sphincter. Esophageal peristaltic contractions were induced by swallow and short- (1-s) and long-train (10-s) vagal stimulation (VS). Galanin (1 nmol/kg) inhibited the amplitude of swallow-induced peristaltic contractions and increased peristaltic velocity by enlarging the latency periods in the upper part of the esophagus and reducing them in the lower part. Galinin nearly abolished esophageal contractions caused by short-train VS at 5 Hz and inhibited the contractions at 10 Hz. Galanin increased latency periods induced by short-train VS with little change in the velocity of peristalsis and reduced the amplitude of both A (cholinergic) and B (noncholinergic) contractions due to long-train VS. However, the decrease in amplitude of B contractions was more marked. Galantide (3 nmol/kg) antagonized the inhibitory action of exogenous galanin on esophageal contractions elicited by short-train VS, but by itself galantide had no significant effect on esophageal contractions. In conclusion, exogenous galanin inhibits the amplitude of swallow induced peristaltic contractions and converts them into nonperistaltic contractions by inhibiting both the cholinergic and noncholinergic components. PMID- 11005759 TI - NK-1 receptor desensitization and neutral endopeptidase terminate SP-induced pancreatic plasma extravasation. AB - Substance P (SP) induces plasma extravasation and neutrophil infiltration by activating the neurokinin-1 receptor (NK1-R). We characterized the mechanisms regulating this response in the rat pancreas. Anesthetized rats were continuously infused with SP, and plasma extravasation was quantified using Evans blue (EB) dye. Continuous infusion of SP (8 nmol. kg(-1). h(-1)) resulted in a threshold increase in EB at 15 min, a peak effect at 30 min (150% increase), and a return to baseline by 60 min. The NK1-R antagonist CP-96,345 blocked SP-induced plasma extravasation. After 60 min, the NK1-R was desensitized to agonist challenge. Resensitization was first detected at 20 min and increased until full recovery was seen at 30 min. Inhibition of the cell-surface protease neutral endopeptidase (NEP) by phosphoramidon potentiated the effect of exogenous SP; therefore endogenous NEP attenuates SP-induced plasma extravasation. Thus the continuous infusion of SP stimulates plasma extravasation in the rat pancreas via activation of the NK1-R, and these effects are terminated by both desensitization of the NK1 R and the cell-surface protease NEP. PMID- 11005760 TI - Identification of gamma-aminobutyric acid receptor subunit types in human and rat liver. AB - GABA is a potent inhibitory neurotransmitter that binds to heterooligomeric receptors in the mammalian brain. In a previous study, we documented specific GABA binding to isolated rat hepatocytes that resulted in inhibition of hepatocyte proliferation. The purpose of the present study was to define the nature of hepatic GABA(A) receptors and to document their expression during rapid liver growth (after partial hepatectomy). PCRs with gene-specific primers derived from published sequences were performed with Marathon-ready human and rat liver cDNA. Two GABA(A) receptor subunit types (beta3 and epsilon) were expressed in human liver and one subunit type (beta3) in rat liver. PCR amplification of the human GABA(A) receptorbeta3-subunit produced a single product (molecular mass 53 59 kDa). In the case of the epsilon-subunit, two PCR products were identified. After partial hepatectomy, GABA(A) receptorbeta3-subunit expression inversely correlated with regenerative activity (r = -0.527, P = 0.006). In conclusion, these results indicate that in the human liver GABA(A) receptors consist of the beta3- and epsilon-subunit types, whereas in the rat liver only the beta3-subunit type is expressed. The results also support the hypothesis that GABAergic activity serves to maintain hepatocytes in a quiescent state. PMID- 11005761 TI - Nitrergic and purinergic regulation of the rat pylorus. AB - The role of nitric oxide (NO) and ATP in the regulation of nonadrenergic, noncholinergic (NANC) inhibitory transmission in the pylorus remains unclear. In the presence of atropine and guanethidine, electric field stimulation induced NANC relaxations in a frequency-dependent manner (1-20 Hz) in the rat pylorus. NANC relaxations were significantly inhibited by N(G)-nitro-L-arginine methyl ester (L-NAME; 10(-4) M). P(2X) purinoceptor antagonist pyridoxal phosphate-6 azophenyl-2',4'-disulfonic acid (PPADS; 3 x 10(-5) M) and P(2Y) purinoceptor antagonist reactive blue 2 (2 x 10(-5) M) had no effect on NANC relaxations. However, the combined administration of L-NAME and PPADS, but not reactive blue 2, evoked greater inhibitory effects on NANC relaxation than that evoked by L NAME alone. alpha-Chymotrypsin and vasoactive intestinal polypeptide antagonist did not affect NANC relaxations. ATP (10(-5)-10(-3) M) and P(2X) purinoceptor agonist alpha, beta-methyleneadenosine 5'-triphosphate (10(-7)-10(-5) M), but not P(2Y) purinoceptor agonist 2-methylthioadenosine 5'-triphosphate (10(-7)-10(-5) M), induced muscle relaxations in a dose-dependent manner, and relaxations were significantly reduced by PPADS and unaffected by TTX. These studies suggest that NO and ATP act in concert to mediate NANC relaxation of the rat pylorus. ATP induced relaxation appears to be mediated by P(2X) purinoceptors located on smooth muscle cells. PMID- 11005762 TI - Cubilin expression and posttranslational modification in the canine gastrointestinal tract. AB - Cubilin is an endocytic receptor of the apical brush border membrane that is essential for intrinsic factor-mediated cobalamin absorption in small intestine. However, cubilin is more highly expressed in kidney and yolk sac, and recent molecular characterization of the receptor has focused on these tissues. The aim of this investigation was to examine tissue-specific cubilin expression and posttranslational modifications with an emphasis on the gastrointestinal tract. Intrinsic factor-cobalamin binding activity, cubilin immunoreactivity, and cubilin mRNA levels were determined in multiple segments of canine gastrointestinal mucosa and other tissues. These aspects of cubilin expression varied in parallel, suggesting that the major determinant of regional cubilin expression in the gastrointestinal tract is modulation of cubilin mRNA. Cell fractionation indicated that ileal cubilin is not strongly membrane associated. An approximately 185-kDa brush border specific and two >400-kDa precursor forms of cubilin were identified. Asparagine-linked oligosaccharide modifications characterized by differential glycosidase digestion of affinity-purified cubilin from ileal mucosa and renal cortex differed, but ileal and renal intracellular cubilin comigrated on SDS-PAGE at approximately 400 kDa after oligosaccharide removal, thus reconciling previous conflicting size estimates of the cubilin polypeptide. PMID- 11005763 TI - Rotavirus alters paracellular permeability and energy metabolism in Caco-2 cells. AB - Rotaviruses infect epithelial cells of the small intestine, but the pathophysiology of the resulting severe diarrhea is incompletely understood. Histological damage to intestinal epithelium is not a consistent feature, and in vitro studies showed that intestinal cells did not undergo rapid death and lysis during viral replication. We show that rotavirus infection of Caco-2 cells caused disruption of tight junctions and loss of transepithelial resistance (TER) in the absence of cell death. TER declined from 300 to 22 Omega. cm(2) between 8 and 24 h after infection and was accompanied by increased transepithelial permeability to macromolecules of 478 and 4,000 Da. Distribution of tight junction proteins claudin-1, occludin, and ZO-1 was significantly altered during infection. Claudin 1 redistribution was notably apparent at the onset of the decline in TER. Infection was associated with increased production of lactate, decreased mitochondrial oxygen consumption, and reduced cellular ATP (60% of control at 24 h after infection), conditions known to reduce the integrity of epithelial tight junctions. In conclusion, these data show that rotavirus infection of Caco-2 intestinal cells altered tight junction structure and function, which may be a response to metabolic dysfunction. PMID- 11005765 TI - Mechanism(s) of butyrate transport in Caco-2 cells: role of monocarboxylate transporter 1. AB - The short-chain fatty acid butyrate was readily taken up by Caco-2 cells. Transport exhibited saturation kinetics, was enhanced by low extracellular pH, and was Na(+) independent. Butyrate uptake was unaffected by DIDS; however, alpha cyano-4-hydroxycinnamate and the butyrate analogs propionate and L-lactate significantly inhibited uptake. These results suggest that butyrate transport by Caco-2 cells is mediated by a transporter belonging to the monocarboxylate transporter family. We identified five isoforms of this transporter, MCT1, MCT3, MCT4, MCT5, and MCT6, in Caco-2 cells by PCR, and MCT1 was found to be the most abundant isoform by RNase protection assay. Transient transfection of MCT1, in the antisense orientation, resulted in significant inhibition of butyrate uptake. The cells fully recovered from this inhibition by 5 days after transfection. In conclusion, our data showed that the MCT1 transporter may play a major role in the transport of butyrate into Caco-2 cells. PMID- 11005764 TI - Separate pathways for cellular uptake of ferric and ferrous iron. AB - Separate pathways for transport of nontransferrin ferric and ferrous iron into tissue cultured cells were demonstrated. Neither the ferric nor ferrous pathway was shared with either zinc or copper. Manganese shared the ferrous pathway but had no effect on cellular uptake of ferric iron. We postulate that ferric iron was transported into cells via beta(3)-integrin and mobilferrin (IMP), whereas ferrous iron uptake was facilitated by divalent metal transporter-1 (DMT-1; Nramp 2). These conclusions were documented by competitive inhibition studies, utilization of a beta(3)-integrin antibody that blocked uptake of ferric but not ferrous iron, development of an anti-DMT-1 antibody that blocked ferrous iron and manganese uptake but not ferric iron, transfection of DMT-1 DNA into tissue culture cells that showed enhanced uptake of ferrous iron and manganese but neither ferric iron nor zinc, hepatic metal concentrations in mk mice showing decreased iron and manganese but not zinc or copper, and data showing that the addition of reducing agents to tissue culture media altered iron binding to proteins of the IMP and DMT-1 pathways. Although these experiments show ferric and ferrous iron can enter cells via different pathways, they do not indicate which pathway is dominant in humans. PMID- 11005766 TI - Systemic lipopolysaccharide influences rectal sensitivity in rats: role of mast cells, cytokines, and vagus nerve. AB - Intraperitoneal lipopolysaccharide (LPS) produces somatic hyperalgesia, releases interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha), and activates vagal afferents. The aim of this study was to evaluate the effect of peripheral LPS on rectal sensitivity and to specify the mechanisms involved. Abdominal muscle contractions were recorded in conscious rats equipped with intramuscular electrodes. Rectal distension (RD) was performed at various times after LPS or experimental treatments. In controls, RD significantly increased the number of abdominal contractions from a threshold volume of distension of 0.8 ml. At the lowest volume (0.4 ml), this number was increased after administration of LPS (3, 9, and 12 h later), recombinant human IL-1beta (from 3 to 9 h), recombinant bovine TNF-alpha (from 6 to 9 h), and BrX-537A (from 6 to 12 h), a mast cell degranulator. The effect of LPS was reduced by doxantrazole, Lys-D-Pro-Thr, and soluble recombinant TNF receptor. Vagotomy selectively amplified the response to LPS. We conclude that, in vivo, intraperitoneal LPS lowers visceral pain threshold (allodynia) through a mechanism involving mast cell degranulation and IL-1beta and TNF-alpha release and that the vagus nerve may exert a tonic protective role against LPS-induced rectal allodynia. PMID- 11005767 TI - Role of cyclooxygenase-2 in Helicobacter pylori- induced gastritis in Mongolian gerbils. AB - Cyclooxygenase (COX)-2 expression is induced in the gastric mucosa of Helicobacter pylori-infected patients, but its role remains unclear. We examined the effects of NS-398 and indomethacin on gastric pathology in H. pylori-infected Mongolian gerbils. COX-1 was detected in both normal and H. pylori-infected mucosa, whereas COX-2 was expressed only in the infected mucosa. PGE(2) production was elevated by H. pylori infection, and the increased production was reduced by NS-398, which did not affect PGE(2) production in normal mucosa. Indomethacin inhibited PGE(2) production in both normal and infected mucosa. Hemorrhagic erosions, neutrophil infiltration, lymphoid follicles, and epithelium damage were induced by H. pylori infection. NS-398 and indomethacin aggravated these pathological changes but did not increase viable H. pylori number. H. pylori-increased production of neutrophil chemokine and interferon-gamma was potentiated by NS-398 and indomethacin. Neither NS-398 nor indomethacin caused any pathological changes or cytokine production in normal animals. These results indicate that COX-2 as well as COX-1 might play anti-inflammatory roles in H. pylori-induced gastritis. PMID- 11005768 TI - Transport of procainamide via H(+)/tertiary amine antiport system in rabbit intestinal brush-border membrane. AB - Transport characteristics of procainamide in the brush-border membrane isolated from rabbit small intestine were studied by a rapid-filtration technique. Procainamide uptake by brush-border membrane vesicles was stimulated by an outward H(+) gradient (pH(in) = 6.0, pH(out) = 7.5) against a concentration gradient (overshoot phenomenon), and this stimulation was reduced when the H(+) gradient was subjected to rapid dissipation by the presence of a protonophore, FCCP. An outward H(+) gradient-dependent procainamide uptake was not caused by H(+) diffusion potential. The initial uptake of procainamide was inhibited by other tertiary amines with N-dimethyl or N-diethyl moieties in their structures, such as triethylamine, dimethylaminoethyl chloride, and diphenhydramine, but not by tetraethylammonium and thiamine. Furthermore, procainamide uptake was stimulated by preloading the vesicles with these tertiary amines (trans stimulation effect), indicating the existence of a specific transport system for tertiary amines. These findings indicate that procainamide transport in the intestinal brush-border membrane is mediated by the H(+)/tertiary amine antiport system that recognizes N-dimethyl or N-diethyl moieties in the structures of tertiary amines. PMID- 11005769 TI - Role of gut-enriched Kruppel-like factor in colonic cell growth and differentiation. AB - Cancer cells differ from normal cells in many aspects, including hyperproliferation and loss of differentiation. Recent research has focused on the role of transcription factors in regulating abnormal cell growth. Gut enriched Kruppel-like factor (GKLF) is a newly identified eukaryotic zinc finger protein expressed extensively in the gastrointestinal tract. In the current study, we demonstrated that GKLF mRNA levels were significantly decreased in the dysplastic epithelium of the colon, including adenomatous polyp and cancer. GKLF immunostains in the normal colon were higher at the surface epithelium and gradually decreased toward the crypt, but this gradient was not present in the adenomatous and cancerous mucosa. Constitutive overexpression of GKLF DNA in a human colonic adenocarcinoma cell line (HT-29) decreased [(3)H]thymidine incorporation, whereas suppression of GKLF gene increased DNA synthesis, indicating that downregulation of the GKLF gene might contribute to cellular hyperproliferation. Cyclin D1 (CD1) protein level and CD1-associated kinase activity were decreased in HT-29 cell overexpressed GKLF cDNA, and CD1 promoter activity was profoundly suppressed by GKLF. When HT-29 cells were cultured in the presence of sodium butyrate, GKLF mRNA levels increased as cells acquired more differentiated phenotypes. These results suggest that GKLF plays an important role in regulating cell growth and differentiation in the colonic epithelium and that downregulation of GKLF expression may cause colonic cells to become hyperproliferative. Furthermore, GKLF appears to be a transcriptional repressor of the CD1 gene. PMID- 11005770 TI - Cell transplantation causes loss of gap junctions and activates GGT expression permanently in host liver. AB - Cell transplantation into hepatic sinusoids, which is necessary for liver repopulation, could cause hepatic ischemia. To examine the effects of cell transplantation on host hepatocytes, we transplanted Fisher 344 rat hepatocytes into syngeneic dipeptidyl peptidase IV-deficient rats. Within 24 h of cell transplantation, areas of ischemic necrosis, along with transient disruption of gap junctions, appeared in the liver. Moreover, host hepatocytes expressed gamma glutamyl transpeptidase (GGT) extensively, which was observed even 2 years after cell transplantation. GGT expression was not associated with alpha-fetoprotein activation, which is present in progenitor cells. Increased GGT expression was apparent after transplantation of nonparenchymal cells and latex beads but not after injection of saline, fragmented hepatocytes, hepatocyte growth factor, or turpentine. Some host hepatocytes exhibited apoptosis, as well as DNA synthesis, between 24 and 48 h after cell transplantation. Changes in gap junctions, GGT expression, DNA synthesis, and apoptosis after cell transplantation were prevented by vasodilators. The findings indicated the onset of ischemic liver injury after cell transplantation. These hepatic perturbations must be considered when transplanted cells are utilized as reporters for biological studies. PMID- 11005771 TI - p53-dependent acinar cell apoptosis triggers epithelial proliferation in duct ligated murine pancreas. AB - The mechanisms linking acinar cell apoptosis and ductal epithelial proliferation remain unknown. To determine the relationship between these events, pancreatic duct ligation (PDL) was performed on p53(+/+) and p53(-/-) mice. In mice bearing a wild-type p53 allele, PDL resulted in upregulation of p53 protein in both acinar cells and proliferating duct-like epithelium. In contrast, upregulation of Bcl-2 occurred only in duct-like epithelium. Both p21(WAF1/CIP1) and Bax were also upregulated in duct-ligated lobes. After PDL in p53(+/+) mice, acinar cells underwent widespread apoptosis, while duct-like epithelium underwent proliferative expansion. In the absence of p53, upregulation of p53 target genes and acinar cell apoptosis did not occur. The absence of acinar cell apoptosis in p53(-/-) mice also eliminated the proliferative response to duct ligation. These data demonstrate that PDL-induced acinar cell apoptosis is a p53-dependent event and suggest a direct link between acinar cell apoptosis and proliferation of duct like epithelium in duct-ligated pancreas. PMID- 11005772 TI - Evaluation of multiple-point measurement of sphincter of Oddi motility in the Australian brush-tailed possum. AB - Manometric assembly diameter is a major limitation on the number of perfused manometric recording points for recordings from the sphincter of Oddi (SO). We evaluated novel polyimide manometric assemblies whereby four recording channels were incorporated in an overall assembly diameter of 0.8 mm. Over the very low range of perfusion rates tested (0.005-0.04 ml/min), the assemblies had pressure offsets attributable to water perfusion from 2 to 23 mmHg and pressure rise rates from 20 to 163 mmHg/s. In six anesthetized Australian brush-tailed possums, manometric recordings from the SO showed a significant reduction in the recorded peak amplitude of pressure waves with perfusion rates below 0.02 ml/min. The pressure profile of the sphincter was found to be asymmetric, and phasic wave propagation patterns were complex (antegrade 35.6%, "mixed" 64.4%). In conclusion, accurate multipoint SO manometry in the possum can be performed with micromanometric assemblies at very low perfusion rates to give a more complete understanding of SO mechanics. These methods are also potentially applicable to perfusion manometry in other small laboratory animals such as mice. PMID- 11005773 TI - Systematic review of the cost effectiveness of prophylactic treatments in the prevention of gastropathy in patients with rheumatoid arthritis or osteoarthritis taking non-steroidal anti-inflammatory drugs. AB - A systematic review on the cost effectiveness of prophylactic treatments of non steroidal anti-inflammatory drug (NSAID) induced gastropathy in patients with osteoarthritis or rheumatoid arthritis was conducted. Two reviewers conducted the literature search and the review. Both full and partial economic evaluations published in English, Dutch, or German were included. The criteria list published in the textbook of Drummond was used to determine the quality of the economic evaluations. The methodological quality of three randomised controlled trials (RCTs) in which the economic evaluations obtained probability estimates of NSAID induced gastropathy and adverse events was assessed by a list of internal validity criteria. The conclusions were based on a rating system consisting of four levels of evidence. Ten economic evaluations were included; three were based on RCTs. All evaluations studied misoprostol as prophylactic treatment: in one evaluation misoprostol was studied as a fixed component in a combination with diclofenac (Arthrotec). All economic evaluations comprised analytical studies containing a decision tree. The three trials were of high methodological quality. Nine economic evaluations were considered high quality and one economic evaluation was considered of low methodological quality. There is strong evidence (level "A") that the use of misoprostol for the prevention of NSAID induced gastropathy is cost effective, and limited evidence (level "C") that the use of Arthrotec is cost effective. Although the levels of evidence used in this review are arbitrary, it is believed that a qualitative analysis is useful: quantitative analyses in this field are hampered by the heterogeneity of economic evaluations. Existing criteria to evaluate the methodological quality of economic evaluations may need refinement for use in systematic reviews. PMID- 11005774 TI - Teaching the teachers: ways of improving teaching and identifying areas for development. PMID- 11005776 TI - Standing committee on epidemiology and health services research: past, present, and future PMID- 11005777 TI - Monocyte chemoattractant protein 1 (MCP-1) in temporal arteritis and polymyalgia rheumatica. AB - OBJECTIVE: To examine the localisation of monocyte chemoattractant protein 1 (MCP 1) in the inflamed vessel wall in temporal arteritis (TA) and to measure MCP-1 in plasma both in patients with TA and patients with polymyalgia rheumatica (PMR). METHODS: By immunohistochemical techniques MCP-1 was localised to the vessel wall in patients with TA. In TA, PMR, and healthy controls MCP-1 was quantified by enzyme linked immunosorbent assay (ELISA) in plasma. RESULTS: MCP-1 was localised to the majority of mononuclear cells, some smooth muscle cells, and giant cells in the arterial biopsy specimens from 12 patients with histologically verified TA. In all sections, including the vasa vasorum, the endothelium stained positive. In the intima 73% (range 57-91%), in the media 49% (range 32-67%), and in the adventitia 74% (range of 62-91%) of all cells stained positive. In plasma MCP-1 was significantly raised in untreated TA (n=33) and untreated PMR (n=27) compared with healthy controls (n=12). Untreated TA plasma levels of MCP-1 (mean 391 pg/ml (range 82-778 pg/ml)) were similar to untreated PMR plasma levels (mean 402 pg/ml (range 29-1153 pg/ml)), and no significant difference was found between the two groups of patients. In both patients with TA and patients with PMR no correlation was found between the plasma level of MCP-1 and the erythrocyte sedimentation rate, haemoglobin concentration, and CD4/CD8 ratio. CONCLUSIONS: These results show that MCP-1 plays a part in the disease processes of TA and PMR. PMID- 11005775 TI - Background for studies on the treatment of male osteoporosis: state of the art. AB - Male osteoporosis represents an important, although long underestimated, public health problem. Both in men and in women aging is accompanied by continuous bone loss and by an exponential increase in the incidence of osteoporotic fracture, with a female to male incidence ratio of about 2 to 3 to 1 in the elderly for hip and vertebral fractures. Morbidity after osteoporotic fractures appears to be more serious and mortality more common in men than in women. To date, no single treatment has been proved to be effective and safe in published prospective studies. The present report, based on a systematic search of the literature on male osteoporosis, summarises the state of the art on the clinical consequences of male osteoporosis and its risk factors, in relation to the present state of knowledge about female osteoporosis. This constitutes the background for the design of rational clinical development strategies for therapeutic interventions in male osteoporosis. From this review of the literature it is apparent that notwithstanding the existing sex differences in pathophysiology of osteoporosis and the difference in age-specific incidence of osteoporotic fractures, there are also important similarities between osteoporosis in women and men. The higher incidence of fracture in women than in men results from quantitative differences in risk factors rather than from different risk factors. Even though there are sex differences in bone geometry, incidence of fracture seems to be similar in men and women for a same absolute areal bone mineral density. However, the lack of data on the changes in fracture rates in men resulting from pharmacological intervention, leading to changes in bone mineral density or bone turnover, remains the main limitation for extrapolation of established treatment outcomes from women to men. PMID- 11005778 TI - Effect of thrombin inhibition on synovial inflammation in antigen induced arthritis. AB - OBJECTIVE: To determine the effect of the thrombin inhibitor, hirudin, on the pathogenesis of murine antigen induced arthritis (AIA). METHODS: AIA was induced by intra-articular injection of methylated bovine serum albumin in the knee joints of previously immunised mice. Hirudin (injected subcutaneously 3 x 200 microg/mouse/day) was given over 13 days, starting three days before arthritis onset, and its anticoagulant effect monitored by clotting times. Arthritis severity was evaluated by technetium-99m ((99m)Tc) uptake in the knee joints and by histological scoring. In addition, intra-articular fibrin deposition was examined by immunohistochemistry, and synovial cytokine mRNA expression measured by RNase protection. RESULTS: Joint inflammation, measured by (99m)Tc uptake, was significantly reduced in hirudin treated mice at days 7 and 10 after arthritis onset. Histologically, synovial thickness was markedly decreased in hirudin treated mice compared with untreated ones. By contrast, no difference in articular cartilage proteoglycan content was found between both groups. Intra articular fibrin deposition and synovial interleukin 1beta mRNA levels, were slightly reduced ( approximately 20%) in arthritic joints from hirudin treated mice compared with untreated ones at day 10 of AIA. CONCLUSION: Hirudin reduces joint inflammation associated with AIA by fibrin-dependent and independent mechanisms. PMID- 11005779 TI - A population study of factors associated with general practitioner consultation for non-inflammatory musculoskeletal pain. AB - OBJECTIVE: To investigate factors associated with visiting a general practitioner (GP) for non-inflammatory musculoskeletal pain, and to examine whether these factors were affected by duration (chronic v non-chronic) or location (widespread v regional) of pain. METHODS: From a cross sectional postal survey of 20 000 (response rate 59%) randomly selected adults in two counties of Norway, 6408 subjects who had experienced musculoskeletal pain during the past month were included. Patients who reported inflammatory rheumatic diagnoses made by a doctor were excluded. RESULTS: 2909 (45%) had consulted a GP for their musculoskeletal pain during the past 12 months. The odds of consulting were significantly increased by being a woman, by having a higher age and lower education, and by being a pensioner or on sick leave. Patients with widespread pain were more likely to consult than those with regional pain, as were patients with chronic compared with non-chronic pain. Greater than median pain intensity was the factor most prominently associated with consultation for men (odds ratio (OR)=2.4; 95% confidence interval (95% CI) 2.0 to 2.9) and for women (OR=2.6; 95% CI 2.3 to 2.9). Overall, consultation was significantly associated with mental distress for women but not for men. Subgroup analyses showed that consultation for chronic pain was significantly associated with greater than median mental distress for both women (OR=1.3; 95% CI 1.1 to 1.6) and men (OR=1.2; 95% CI 1. 0 to 1.4), whereas consultation for non-chronic pain was not. CONCLUSION: The results show that about half of the patients with musculoskeletal pain consult a general practitioner (GP) each year, that demographic factors are associated with consulting, and that the role of mental distress for consulting a GP varies with duration of pain. PMID- 11005781 TI - Pregnancy in patients with Wegener's granulomatosis: report of five cases in three women. AB - Five cases of pregnancy occurring in three women with previously diagnosed Wegener's granulomatosis are described. The disease was diffuse in one case and localised in the other. Initial treatment consisted of a combination of corticosteroids and intravenous cyclophosphamide in two women, and methotrexate in one. Four pregnancies ended in live births despite pre-eclampsia in two cases. One therapeutic abortion was induced because of encephalocele. Comparable reported cases were reviewed to examine the implications of immunosuppressive treatment on the fetus. A relapse occurred during pregnancy in 40% of the cases, but in 25% if only pregnancies beginning during inactive disease were taken into account. No other indicator for maternal and fetal outcome was obvious. Pregnancy should be planned after complete disappearance of disease activity. In the case of a relapse a combination of immunosuppressive drugs and corticosteroids should be chosen rather than corticosteroids alone because the outcome of pregnancy is poor in cases of undertreatment. Prematurity remains common. PMID- 11005780 TI - Socioeconomic deprivation and rheumatoid disease: what lessons for the health service? ERAS Study Group. Early Rheumatoid Arthritis Study. AB - OBJECTIVE: To assess how socioeconomic deprivation influences the presentation, treatment, and outcome of patients with rheumatoid arthritis (RA). METHODS: Three year follow up of 869 consecutive patients with RA from nine hospital rheumatology clinics, with patients categorised by the Carstairs deprivation score of their enumeration district of residence. Outcomes included Health Assessment Questionnaire (HAQ), joint and pain scores, grip strength, functional grade, radiological evidence of bony erosions, and medical/surgical interventions. RESULTS: Patients from more deprived enumeration districts presented with more severe disease as judged by the HAQ score and joint scores. An increase from the 5th to the 95th centile of the Carstairs distribution was associated with an odds ratio of 1.87 (95% confidence interval (95% CI) 1.31 to 2.66) for an above-median HAQ score and 1.77 (95% CI 1.23 to 2.54) for an above median joint score. Statistically non-significant deprivation trends were seen with erythrocyte sedimentation rate, pain score, and grip strength. By three years, despite no important differences in clinical management, socioeconomic differentials had worsened or remained unchanged such that clear deprivation trends were then seen in HAQ (p=0.002) and joint scores (p=0.001), in grip strength (p=0. 008), and in functional grade (p=0.003). The association between deprivation and HAQ at three years was present after adjustment for age, sex, treatment centre, and HAQ at presentation (adjusted odds ratio 1.74, 95% CI 1.1 to 2.74). CONCLUSIONS: Socioeconomic deprivation was associated with a worse clinical course of rheumatoid disease, and this effect was already apparent at presentation, but not with systematic differentials in its treatment. This suggests that individual susceptibility and lifestyle factors contribute to socioeconomic differentials in outcome, an observation that has implications for clinical management. PMID- 11005782 TI - Data driven attempt to create a clinical algorithm for identification of women with rheumatoid arthritis at high risk of osteoporosis. AB - OBJECTIVES: To examine relations between osteoporosis and low bone mass and demographic and clinical variables in patients with rheumatoid arthritis (RA), in an attempt to develop a data driven clinical tool for identification of patients at high risk of osteoporosis. METHODS: All patients were recruited from a county based register and were examined cross sectionally with a variety of clinical and health status measures as well as bone density measures (anteroposterior spine L2 4, total hip, and femoral neck). Associations between osteoporosis (T score < or = -2.5SD) and low bone mass (T score < or = -1SD), on the one hand, and demographic and clinical measures, on the other, were examined bivariately and by logistic regression analyses. RESULTS: 394 patients with a mean age of 54.8 years were examined. The percentages having osteoporosis/low bone mass were 16.8/45.8, 14.7/54.5 and 14.7/55.5 in spine L2-4, total hip, and femoral neck, respectively. Osteoporosis and low bone mass were bivariately related to age, body mass index (BMI), disease duration, disease process measures, presence of deformed joints, physical disability, current use of corticosteroids, and history of non-vertebral fracture. In multivariate analyses, age >60 years, low BMI, and current use of corticosteroids were consistently related to osteoporosis and to low bone mass at all sites. The presence of deformed joints was associated with osteoporosis at the total hip, and a history of previous non-vertebral fracture with osteoporosis at the femoral neck. The Modified Health Assessment Questionnaire (MHAQ) > or = 1.5 and non-vertebral fracture were also independently associated with low bone mass at the hip. The logistic regression analyses models could, however, only predict osteoporosis with a sensitivity of about 50-60% and a specificity of 80 90% at the various measurement sites, and low bone mass with a sensitivity and specificity of about 70%. CONCLUSION: Consideration of demographic and disease markers may be of some help in predicting presence of osteoporosis or low bone mass, but a combination of markers cannot be used as a clinical tool with sufficient sensitivity and specificity for the identification of osteoporosis or low bone mass in patients with RA. PMID- 11005783 TI - Motor performance of the hand in patients with rheumatoid arthritis. AB - OBJECTIVES: To examine the motor performance of the hand in a sample of patients with rheumatoid arthritis (RA). SUBJECTS: The patient group comprised 21 (two men, 19 women) patients with RA. Twenty one control subjects matched for age and sex were selected from a larger reference group, which had been drawn from the local population. METHODS: The measured motor performance aspects were simple reaction time, choice reaction time, speed of movement, finger tapping speed, and coordination (that is, speed of movement/accuracy). Results were compared for age and sex matched pairs. The measurements were made with the Human Performance Measurement/Basic Elements of Performance system, which is a multifunctional system designed to measure different motor aspects of the hands, including reaction time, movement speed, tapping speed, and coordination. RESULTS: A comparison of the results for the patient and control groups indicated that the motor functions of patients with RA were impaired in all the measured aspects (with the exception of the index finger tapping test). The difference between the groups varied between 11% and 21% for the reaction time tasks, between 12% and 18% for the speed of movement tasks, and between 15% and 17% for the coordination task. CONCLUSION: Based on the results of our research, it seems that RA decreases some motor performance functions of the hand expressed as simple reaction time, choice reaction time, speed of movement, and coordination. The changes were emphasised in movements performed with several joints. PMID- 11005784 TI - Bilateral hip osteonecrosis: influence of hip size on outcome. AB - OBJECTIVE: To measure the volume of osteonecrosis in 30 patients with disease in both hips (one side with collapse and the other without at Ficat stage I). METHOD: The volume of the osteonecrosis was measured by magnetic resonance imaging. RESULTS: The progression to collapse was influenced by the size of the lesion in each patient. The first collapsed hip was that with the largest volume of osteonecrosis. CONCLUSION: The volume of the osteonecrotic lesion on the second hip (stage I without collapse) is a good predictor of the time to collapse of this second hip. PMID- 11005785 TI - Expression of epidermal growth factor and transforming growth factor alpha in interfacial membranes retrieved at revision total hip arthroplasty. AB - BACKGROUND: The interfacial membrane between bone and implant has been shown to be a key tissue in the process of aseptic loosening of total hip arthroplasty. The cells within the interfacial membrane produce numerous inflammatory mediators which, through complex mechanisms, cause periprosthetic osteolysis and aseptic loosening. Both epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha) have similar biological functions. They have been found to stimulate bone resorption. OBJECTIVE: To investigate the presence, cellular localisation, and extent of expression of EGF and TGFalpha in interfacial membrane retrieved from revision total hip arthroplasty and compare it with that in synovial membrane from primary total hip arthroplasty. METHODS: Ten interfacial membranes and 10 synovial membranes were stained with avidin-biotin peroxidase complex for EGF and TGFalpha. The staining process was done using the Lab Vision Autostainer. The results were measured by a semiautomatic VIDAS image analysis system. RESULTS: Immunoreactivity for both EGF and TGFalpha was found in the endothelial cells of blood vessels, macrophages, and fibroblasts, both in interfacial membranes and synovial membranes. However, the number of EGF (980 (370)) and TGFalpha (1070 (360)) positive cells per mm(2) was greater in interfacial membranes than in the synovial membranes (220 (200), 270 (100); p<0.01). CONCLUSION: It is suggested that owing to their increased expression in interfacial membrane, EGF and TGFalpha may have an important pathogenetic role in stimulating periprosthetic bone resorption in aseptic loosening of total hip arthroplasty. PMID- 11005786 TI - Serum levels of YKL-40 and C reactive protein in patients with hip osteoarthritis and healthy subjects: a cross sectional study. AB - BACKGROUND: YKL-40 is a 40 kDa glycoprotein secreted by chondrocytes and synoviocytes. It has been suggested that it is a surrogate marker of synovial inflammation and joint destruction in rheumatoid arthritis (RA) and osteoarthritis (OA) and related to C reactive protein (CRP) serum levels in RA. OBJECTIVE: To study serum levels of YKL-40 in patients with hip OA and its relation with CRP. METHODS: YKL-40 and CRP were assayed in serum samples from 45 patients (24 women, 21 men, mean age 65) with symptomatic OA of the hip and 33 healthy controls. YKL-40 was assayed by immunoassay and CRP by ultrasensitive immunonephelometry. OA severity was assessed by the measurement of joint space width with a computer analysis system of digitised hip radiographs. Statistical analysis was performed to determine correlations between serum markers and radiological joint space width. RESULTS: The mean (standard error) YKL-40 level was 90.3 (8.2) ng/ml in patients with hip OA and 66.9 (8.2) ng/ml in controls (p=0.03). The mean CRP level was 2.93 (3.03) mg/l in OA and 1.40 (1.61) mg/l in controls (p=0.006). The serum levels of YKL-40 and CRP increased with age and were significantly correlated (Spearman test: r(s)=0.42, p=0.005) in patients but not in controls. Neither YKL-40 nor CRP correlated with radiographic joint space width. CONCLUSIONS: Serum YKL-40 was significantly increased in patients with hip OA. The correlation between YKL-40 and CRP suggests that YKL-40 may be a marker of joint inflammation in OA. Longitudinal studies are required to assess the usefulness of YKL-40 in the monitoring of patients with hip OA. PMID- 11005787 TI - Do present damage and health perception in patients with systemic lupus erythematosus predict extent of future damage?: a prospective study. AB - OBJECTIVE: To study whether either initial damage, disease activity, disease duration, age, a drug score, or health status would predict an increase in damage in patients with systemic lupus erythematosus (SLE) within the next three years. METHODS: A three year prospective longitudinal study of a cohort of 141 consecutive patients with SLE attending a specialist lupus outpatient clinic from their first assessment between July 1994 and February 1995. Disease activity was assessed using the BILAG system, health status by the Medical Outcome Survey Short Form 20 with an extra question about fatigue (SF-20+), and damage by the SLICC/ACR Damage Index (SDI). Damage was reassessed three years later. Statistical analysis was carried out using logistic regression analysis (logXact). RESULTS: 133 female and 8 male patients with SLE (97 white subjects, 16 Afro-Caribbeans, 22 Asians, and 6 others) were included. Their mean (SD) age at inclusion was 41.1 (12.5) years and their disease duration 10.2 (6. 3) years. The mean measures at inclusion were: total BILAG 5.2 (range 0-17), total SDI 1.2 (0-7), drug score 1.2 (0-3); SF-20+: physical 58 (0-100), role 54 (0-100), social functioning 71 (0-100), mental health 64 (16-100), health perception 44 (0-100), pain 53 (0-100), fatigue 59 (0-100). Four patients were lost to follow up because they had moved. At three years in 33 patients the total SDI had increased to a mean of 1.5 (0-7) (n=130). Moreover, seven patients had the maximum damage as they had died during the follow up period. The only variables with an independent and significant contribution in predicting damage at three years were the total damage score (odds ratio (OR)=1.46; 95% CI 1.04 to 2.05), and health perception (OR=0.96; 95% CI 0.93 to 0.99) at inclusion. CONCLUSIONS: Of all the variables at inclusion only the total damage score and SF-20+: health perception, significantly predicted an increase in damage, for patients with SLE, three years later. PMID- 11005788 TI - Unusual presentation of familial Mediterranean fever: role of genetic diagnosis. AB - OBJECTIVE: To describe the role of molecular analysis in the diagnosis of an unusual presentation of familial Mediterranean fever (FMF). CASE REPORT: Two patients presenting with prolonged fever without signs and symptoms of serositis are described. FMF was diagnosed by genetic analysis, which disclosed that both patients were homozygous for the M694V mutation of the Mediterranean fever (MEFV) gene. CONCLUSION: Molecular analysis of FMF should complement the investigation of patients with fever of unknown origin. This test enables a definite diagnosis of the disease and may promote the diagnosis and treatment of patients with an unusual or incomplete clinical picture of FMF. PMID- 11005789 TI - Decoding the human genome sequence. AB - The year 2000 is marked by the production of the sequence of the human genome. A 'working draft' of high quality sequence covering 90% of the genome has been determined and a quarter is in finished form, including the first two completed chromosomes. All sequence data from the project is made freely available to the community via the Internet, for further analysis and exploitation. The challenge which lies ahead is to decipher the information. Knowledge of the human genome sequence will enable us to understand how the genetic information determines the development, structure and function of the human body. We will be able to explore how variations within our DNA sequence cause disease, how they affect our interaction with our environment and ultimately to develop new and effective ways to improve human health. PMID- 11005790 TI - Genetics of asthma and allergic disease. AB - Atopic (allergic) asthma is the most common disease of childhood and is strongly genetic in origin. Many genome-wide screens for asthma and its associated traits have now been carried out, and genetic linkage has been consistently identified in several regions. It is probable that these loci contain major genes influencing atopy and asthma. Candidate genes have already been identified from the cytokine cluster on chromosome 5 and the MHC on chromosome 6. These complex regions contain more than one susceptibility locus for allergic disease. Other regions do not contain obvious candidate genes, and positional cloning of these loci is likely to identify novel disease pathways. Parent-of-origin effects are prominent at some of the loci and some also show linkage to other inflammatory immune diseases. Several single gene disorders are associated with allergic disease and on occasion are also linked to the same chromosomal regions. The positional cloning of asthma genes is now feasible. PMID- 11005791 TI - Rett syndrome: a surprising result of mutation in MECP2. AB - The identification of mutations in the gene encoding methyl CpG binding protein 2 (MeCP2) in Rett syndrome represents a major advance in the field. The current model predicts that MeCP2 represses transcription by binding methylated CpG residues and mediating chromatin remodeling. A physical interaction between MeCP2, histone deacetylases and the transcriptional co-repressor Sin3A has been demonstrated, as well as an association of MeCP2 with the basal transcription apparatus. It is unclear, however, whether MeCP2-mediated chromatin remodeling is necessary for transcriptional repression in vivo. Eight recurrent missense and nonsense mutations account for >65% of the mutations identified in Rett syndrome probands, and as predicted from the sporadic nature of the disorder, most mutations are de novo. The severity of the phenotype is likely to reflect the pattern of X chromosome inactivation in relevant tissues, although the type and position of the mutation may also play a role. Although much is known about the biochemical function of MeCP2, the phenotype of Rett syndrome suggests that it plays an unexplored but critical role in development and maintenance of the nervous system. PMID- 11005792 TI - Haemochromatosis: novel gene discovery and the molecular pathophysiology of iron metabolism. AB - The application of molecular genetics to haemochromatosis and experimental mutagenesis in animals has transformed our capacity to investigate the unique physiology of iron homeostasis-a key problem in biology and medicine. The identification of HFE, the principal determinant of adult haemochromatosis (HFE1; OMIM 235200) and TfR2, recently implicated in a rarer form of the inherited disorder (HFE3; OMIM 604250), and the promise of candidate genes for juvenile haemochromatosis (HFE2; OMIM 602390) and neonatal haemochromatosis (OMIM 231100) provide the foundation for important studies into the control mechanism of iron balance in humans. The rare conditions atransferrinaemia (OMIM 209300) and acaeruloplasminaemia (OMIM 604290), each associated with tissue iron overload, have already implicated the iron transport ligand transferrin and the copper transporter caeruloplasmin in the control of iron homeostasis. Gene mapping studies in animal mutants with anaemia due to defects in the uptake or tissue transfer of iron have yielded novel proteins involved in iron transport: DMT1 (brush border transporter of ferrous iron) in the mk/mk mouse, hephaestin (basolateral multi-copper ferroxidase) in the sex-linked anaemic mouse (sla) and ferroportin1 (basolateral iron exporter) in zebrafish weh mutants. The discovery of genes that determine heritable defects of iron absorption and regulation in animals and humans thus holds promise for a complete mechanistic understanding of the molecular pathophysiology of iron metabolism. PMID- 11005793 TI - Transcriptional regulation of Alzheimer's disease genes: implications for susceptibility. AB - In recent years, important progress has been made in uncovering genes implicated in Alzheimer's disease (AD). Three causal genes have been identified in which mutations cause familial presenile AD: the amyloid precursor protein gene and the presenilin 1 and 2 genes. Additionally, the epsilon 4 allele of the apolipoprotein E gene was shown to be a major risk factor for AD. Despite the genetic heterogeneity, all of these genes work through a common mechanism, i. e. increasing the amount and deposition of the amyloid beta peptide (A beta) in brain triggering AD-related neuronal degeneration. Therefore, the levels of A beta and of the factors involved in its production and deposition are important in the neuropathogenesis of AD. Regulation of transcription of AD genes might therefore be an important player in the neurodegenerative process. In this review, we describe the major features of transcriptional regulation of the known AD genes and the implications of variable expression levels on susceptibility to AD. PMID- 11005794 TI - The DNA methyltransferases of mammals. AB - The biological significance of 5-methylcytosine was in doubt for many years, but is no longer. Through targeted mutagenesis in mice it has been learnt that every protein shown by biochemical tests to be involved in the establishment, maintenance or interpretation of genomic methylation patterns is encoded by an essential gene. A human genetic disorder (ICF syndrome) has recently been shown to be caused by mutations in the DNA methyltransferase 3B (DNMT3B) gene. A second human disorder (Rett syndrome) has been found to result from mutations in the MECP2 gene, which encodes a protein that binds to methylated DNA. Global genome demethylation caused by targeted mutations in the DNA methyltransferase-1 (Dnmt1) gene has shown that cytosine methylation plays essential roles in X-inactivation, genomic imprinting and genome stabilization. The majority of genomic 5 methylcytosine is now known to enforce the transcriptional silence of the enormous burden of transposons and retroviruses that have accumulated in the mammalian genome. It has also become clear that programmed changes in methylation patterns are less important in the regulation of mammalian development than was previously believed. Although a number of outstanding questions have yet to be answered (one of these questions involves the nature of the cues that designate sites for methylation at particular stages of gametogenesis and early development), studies of DNA methyltransferases are likely to provide further insights into the biological functions of genomic methylation patterns. PMID- 11005795 TI - Single nucleotide polymorphisms as tools in human genetics. AB - The development of detailed single nucleotide polymorphism (SNP) maps of the human genome coupled with high-throughput genotyping technologies may allow us to unravel complex genetic traits, such as multifactorial disease or drug response, over the next few years. Here we describe the current efforts to identify and characterize the large numbers of SNPs required and discuss the practicalities of association studies for the identification of genes involved in complex traits. PMID- 11005796 TI - Counting cross-overs: characterizing meiotic recombination in mammals. AB - Until recently, most of our understanding of meiotic recombination has come from studies of lower eukaryotes. However, over the past few years several components of the mammalian meiotic recombination pathway have been identified, and new molecular and cytological approaches to the analysis of mammalian meiosis have been developed. In this review, we discuss recent advances in three areas: the application of new techniques to study genome-wide levels of recombination in individual meioses; studies analyzing temporal aspects of the mammalian recombination pathway; and studies linking the genesis of human trisomies to alterations in meiotic exchange patterns. PMID- 11005797 TI - The 22q11 deletion syndromes. AB - DiGeorge syndrome, velocardiofacial syndrome and various other malformations have been described in association with deletions and translocations involving human chromosome 22q11. Many of the structural malformations observed are also seen in animal models of neural crest disruption suggesting that the haplo-insufficiency resulting from the deletion somehow affects this group of cells or their interactions. Over the past few years it has been shown that the deletion predisposes to a range of psychotic conditions prompting the hypothesis that the deleted region may contain a predisposition locus for psychotic illness. The DiGeorge chromosomal region has been entirely sequenced and many of the genes mapping to the deletion interval have been studied in some detail. Despite these efforts, no gene has yet been proved to play a defined role in the pathogenesis of the syndrome. Current efforts are directed at the study of engineered chromosome mouse models which offer the potential to dissect at least some of the developmental pathways disrupted in this intriguing group of malformation syndromes. PMID- 11005798 TI - Homologous recombination as a mechanism for genome rearrangements: environmental and genetic effects. AB - Novel findings over the last 2 years have led to an increased emphasis on homologous recombination (HR) as both a pathway for DNA repair and a cause for genomic rearrangements. Indeed, environmental carcinogens increase the frequency of HR, as can be observed when two copies of a duplicated sequence recombine to delete the intervening sequences. Such HR events between dispersed homologous sequences may result in not only deletions, but also gene duplications or translocations. These types of genomic rearrangement have been observed to be the cause of several different genetic diseases, including cancer. In reflection of this, several genes have been identified that, when mutant, predispose an individual to an increased frequency of cancer. These genes have been shown to be either directly or indirectly involved in HR. In addition, HR is induced by a wide variety of carcinogens, preferentially in proliferating cells. This fits the most current models of recombination and its involvement in reinitiating stalled replication forks. Thus, 'correct' HR repair may act with high fidelity, an important issue for proliferating cells, but in the context of alternative homologous partner sequences, 'aberrant' HR can cause genomic rearrangements with dire consequences. PMID- 11005799 TI - An update on genetic, structural and functional studies of arylamine N acetyltransferases in eucaryotes and procaryotes. AB - Arylamine N:-acetyltransferase (NAT) was first identified as the inactivator of the anti-tubercular drug isoniazid. The enzyme was shown to catalyse the transfer of an acetyl group from acetyl-CoA to the terminal nitrogen of the hydrazine drug. The rate of inactivation of isoniazid was polymorphically distributed in the population and was one of the first examples of pharmacogenetic variation. NAT was identified recently in Mycobacterium tuberculosis and is a candidate for modulating the response to isoniazid. Genome sequences have revealed many homologous members of this unique family of enzymes. The first three-dimensional structure of a member of the NAT family identifies a catalytic triad consisting of aspartate, histidine and cysteine proposed to form the activation mechanism. So far, all procaryotic NATs resemble the human enzyme which acetylates isoniazid (NAT2). Human NAT2 is characteristic of drug-metabolizing enzymes: it is found in liver and intestine. In humans and other mammals, there are up to three different isoenzymes. If only one isoenzyme is present, it is like human NAT1. Human NAT1 and its murine equivalent specifically acetylate the folate catabolite p aminobenzoylglutamate. NAT1 and its murine homologue each have a ubiquitous tissue distribution and are expressed early in development at the blastocyst stage. During murine embryonic development, NAT is expressed in the developing neural tube. The proposed endogenous role of NAT in folate metabolism, and its multi-allelic nature, indicate that its role in development should be assessed further. PMID- 11005800 TI - Zebrafish: bridging the gap between development and disease. AB - The zebrafish has been the model of choice amongst developmental biologists for many years. This small freshwater species offers many advantages to the study of organ and tissue development that are not provided by other model systems. Against this background, modern molecular genetic approaches are being applied to expand the physical and genetic mapping of the zebrafish genome. These approaches complement the large-scale mutagenic screens that have led to the isolation of mutant phenotypes. Some of the phenotypes have been found to resemble human disease states, while mapping and sequencing data have revealed zebrafish genes with significant homology to human disease-causing genes. It is the realization that the zebrafish offers an amenable system for understanding disease, as opposed to development, that underpins this review. The adventitious identification of disease phenotypes amongst zebrafish mutants and the important area of deliberate disease modelling using transgenesis and gene targeting should lead to a better application of the zebrafish as a vertebrate model of human diseases. PMID- 11005801 TI - Animal models of spinal muscular atrophy. AB - Proximal spinal muscular atrophy (SMA) is the second most common autosomal recessive inherited disorder in humans. It is the most common genetic cause of infant mortality. As yet, there is no cure for this neuromuscular disorder which affects the lower motor neurons and proximal muscles of the limbs and trunk. In the last decade, significant advances have been made in understanding this disease, from linkage analysis to isolating the defective gene and identifying its protein product. This review summarizes the most recent advance in SMA research: the development of animal models of the disease, in particular mouse models of SMA. The SMA mice that we describe here present with symptoms similar to those seen in SMA patients. They promise to further the understanding of the molecular basis of this disease and demonstrate the feasibility of using the intact SMN2 gene, found in all SMA patients, as a means of treating this disorder. PMID- 11005802 TI - Animal models for muscular dystrophy: valuable tools for the development of therapies. AB - Since the identification of dystrophin as the causative factor in Duchenne muscular dystrophy, an increasing amount of information on the molecular basis of muscular dystrophies has facilitated the division of these heterogeneous disorders into distinct groups. As more light is being shed on the genes and proteins involved in muscular dystrophy, diagnosis of patients has improved enormously. In addition to naturally occurring animal models, a number of genetically engineered murine models for muscular dystrophy have been generated. These animal models have provided valuable clues to the understanding of the pathogenesis of these disorders. Furthermore, as therapeutic approaches are being developed, mutant animals represent good models in which they can be tested. The present review focuses on the recent advancements of gene transfer-based strategies, with a special emphasis on animal models for Duchenne and limb-girdle muscular dystrophies. PMID- 11005803 TI - Functional interaction of proliferating cell nuclear antigen with MSH2-MSH6 and MSH2-MSH3 complexes. AB - Eukaryotic DNA mismatch repair requires the concerted action of several proteins, including proliferating cell nuclear antigen (PCNA) and heterodimers of MSH2 complexed with either MSH3 or MSH6. Here we report that MSH3 and MSH6, but not MSH2, contain N-terminal sequence motifs characteristic of proteins that bind to PCNA. MSH3 and MSH6 peptides containing these motifs bound PCNA, as did the intact Msh2-Msh6 complex. This binding was strongly reduced when alanine was substituted for conserved residues in the motif. Yeast strains containing alanine substitutions in the PCNA binding motif of Msh6 or Msh3 had elevated mutation rates, indicating that these interactions are important for genome stability. When human MSH3 or MSH6 peptides containing the PCNA binding motif were added to a human cell extract, mismatch repair activity was inhibited at a step preceding DNA resynthesis. Thus, MSH3 and MSH6 interactions with PCNA may facilitate early steps in DNA mismatch repair and may also be important for other roles of these eukaryotic MutS homologs. PMID- 11005804 TI - Insertion or deletion of a single residue in the strut sequence of Dictyostelium myosin II abolishes strong binding to actin. AB - The strut loop, one of the three loops that connects the upper and lower 50K subdomains of myosin, plays a role as a "strut" to keep the relative disposition of the two subdomains. A single residue was either inserted into or deleted from this loop. The insertion or deletion mutation abolished the in vivo motor functions of myosin, as revealed by the fact that the mutant myosins did not complement the phenotypic defects of the myosin-null cells. In vitro studies of purified full-length myosins and their subfragment-1s (S1s) revealed that the insertion mutants virtually lost the strong binding to actin although their motor functions in the absence of actin remained almost normal, showing that only the hydrophobic actin-myosin association was selectively affected by the insertion mutations. Unlike the insertion mutants, the deletion mutant showed defects both in the strong-binding state and the rate-limiting step of ATPase cycle. These results indicate the functional importance of the strut loop in establishing the strong-binding state of myosin and thereby achieving successful power strokes. PMID- 11005805 TI - Both the structure and DNA binding function of the barrier-to-autointegration factor contribute to reconstitution of HIV type 1 integration in vitro. AB - Retroviral integration is mediated by viral preintegration complexes (PICs), and human immunodeficiency virus type 1 (HIV-1) PICs treated with high salt lose their in vitro integration activity. Barrier-to-autointegration factor (BAF) is a host protein that efficiently restores PIC activity, but the mechanism(s) by which BAF participates in HIV-1 integration remains largely unknown. Here we developed a gel shift assay to study BAF DNA binding, and analyzed 14 mutant proteins containing substitutions of conserved residues for binding and PIC reconstitution activities. Although wild-type BAF efficiently bound double stranded DNA, binding to single-stranded DNA, RNA, or an RNA/DNA hybrid was not detected, suggesting that BAF associates with retroviral cDNA relatively late during reverse transcription. Although some of the BAF mutant proteins efficiently bound DNA, others were defective for binding. Mutants that bound DNA efficiently reconstituted HIV-1 integration, even though in one case binding was just 0.2% of wild-type BAF. Although misfolded mutants did not reconstitute integration, a structurally intact DNA binding-defective mutant displayed partial activity at high BAF concentration. We therefore conclude that both BAF protein structure and its DNA binding activity play roles in reconstituting HIV-1 integration in vitro. PMID- 11005806 TI - Conformational and dynamic differences between actin filaments polymerized from ATP- or ADP-actin monomers. AB - Conformational and dynamic properties of actin filaments polymerized from ATP- or ADP-actin monomers were compared by using fluorescence spectroscopic methods. The fluorescence intensity of IAEDANS attached to the Cys(374) residue of actin was smaller in filaments from ADP-actin than in filaments from ATP-actin monomers, which reflected a nucleotide-induced conformational difference in subdomain 1 of the monomer. Radial coordinate calculations revealed that this conformational difference did not modify the distance of Cys(374) from the longitudinal filament axis. Temperature-dependent fluorescence resonance energy transfer measurements between donor and acceptor molecules on Cys(374) of neighboring actin protomers revealed that the inter-monomer flexibility of filaments assembled from ADP-actin monomers were substantially greater than the one of filaments from ATP-actin monomers. Flexibility was reduced by phalloidin in both types of filaments. PMID- 11005807 TI - Substrate-assisted catalysis of the PAR1 thrombin receptor. Enhancement of macromolecular association and cleavage. AB - Platelet activation and aggregation are mediated by thrombin cleavage of the exodomain of the PAR1 receptor. The specificity of thrombin for PAR1 is enhanced by binding to a hirudin-like region (Hir) located in the receptor exodomain. Here, we examine the mechanism of thrombin-PAR1 recognition and cleavage by steady-state kinetic measurements using soluble PAR1 N-terminal exodomains. We determined that the primary role of the PAR1 Hir sequence is to reduce the kinetic barriers to formation of the docked thrombin-PAR1 complex rather than to form high affinity ground-state interactions. In addition, the exosite I-bound Hir motif facilitates the productive interaction of the PAR1 (38)LDPR/SFL(44) sequence with the active site of thrombin. This locking process is the most energetically unfavorable step of the overall reaction. The subsequent irreversible steps of peptide bond cleavage are rapid and allosterically enhanced by the presence of the docked Hir sequence. Furthermore, the C-terminal exodomain product of thrombin cleavage, corresponding to the activated receptor, binds tightly to thrombin. This would suggest that an additional role of the Hir sequence in the thrombin-activated receptor is to sequester thrombin to the platelet surface and modulate cleavage of other platelet receptors such as the PAR4 thrombin receptor, which lacks a functional Hir sequence. PMID- 11005808 TI - The mechanism of heat shock activation of ERK mitogen-activated protein kinases in the interleukin 3-dependent ProB cell line BaF3. AB - We have investigated heat shock stimulation of MAPK cascades in an interleukin 3 dependent cell line, BaF3. Following exposure to 42 degrees C, the stress activated JNK MAPKs were phosphorylated and activated, but p38 MAPKs remained unaffected. Surprisingly, heat shock also activated ERK MAPKs in a potent (>60 fold), delayed (>30 min), and sustained (>/=120 min) manner. These characteristics suggested a novel mechanism of ERK MAPK activation and became the focus of this study. A MEK-specific inhibitor, PD98059, inhibited heat shock ERK MAPK activation by >75%. Surprisingly, a role for Ras in the heat shock response was eliminated by the failure of a dominant-negative Ras(Asn-17) mutant to inhibit ERK MAPK activation and the failure to observe increases in Ras.GTP. Heat shock also failed to stimulate activation of A-, B-, and c-Raf. Instead, a serine/threonine phosphatase inhibitor, okadaic acid, activated ERK MAPK in a similar manner to heat shock. Furthermore, pretreatment with suramin, generally recognized as a broad range inhibitor of growth factor receptors, inhibited both okadaic acid-stimulated and heat shock-stimulated ERK MAPK activity by >40%. Inhibiting ERK MAPK activation during heat shock with PD98059 enhanced losses in cell viability. These results demonstrate Ras- and Raf-independent ERK MAPK activation maintains cell viability following heat shock. PMID- 11005809 TI - Opposite functions for E2F1 and E2F4 in human epidermal keratinocyte differentiation. AB - Proteins of the retinoblastoma family (pRb, p107, and p130) modulate cell proliferation, a function related to their capacity to control the activity of the E2F transcription factor family. The Rb proteins also control cell differentiation in different tissues. We have recently described their involvement in human epidermal keratinocyte differentiation (Paramio, J. M., Lain, S., Segrelles, C., Lane, E. B. , and Jorcano, J. L. (1998) Oncogene 17, 949 957). Here we show that E2F proteins are also involved in this process. We found that E2F1 and E2F4 are expressed differentially during the in vitro differentiation of human epidermal keratinocytes, with the former uniformly present throughout the process, whereas the second is predominantly expressed at the onset of differentiation. This pattern is also observed in human skin by confocal microscopy. Electrophoretic mobility shift assays and immunoprecipitation experiments demonstrated that the complexes formed by E2F1 and E2F4 and Rb family proteins vary throughout in vitro keratinocyte differentiation. In agreement with this observation, several E2F-responsive genes are differentially regulated during this process. To test the functional implications of these observations, we transfected HaCaT keratinocytes with plasmids coding for E2F1 and E2F4. Transfected cells display opposite in vitro differentiation properties. Although E2F1-transfected cells are unable to differentiate, E2F4-transfected cells show an increased differentiation rate compared with Neo-transfected control cells. Our data demonstrate that the differential and coordinated expression and interaction of E2F and Rb proteins modulate the process of epidermal differentiation and provide clear evidence that members of the E2F family of transcription factors play specific and opposite roles during cell differentiation. PMID- 11005810 TI - Cks1 mediates vascular smooth muscle cell polyploidization. AB - Vascular smooth muscle cells (VSMC) at capacitance arteries of hypertensive individuals and animals undergo dramatic polyploidization that contributes toward their hypertrophic phenotype. We report here the identification of a defective mitotic spindle cell cycle checkpoint in VSMC isolated from capacitance arteries of pre-hypertensive rats. These cells demonstrated a high predisposition to polyploidization in culture and failed to maintain cyclin B protein levels in response to colcemid, a mitotic inhibitor. Furthermore, this altered mitotic spindle checkpoint status was associated with the overexpression of Cks1, a Cdc2 adapter protein that promotes cyclin B degradation. Cks1 up-regulation, cyclin B down-regulation, and VSMC polyploidization were evidenced at the smooth muscle of capacitance arteries of genetically hypertensive and Goldblatt-operated rats. In addition, angiotensin II infusion dramatically increased Cks1 protein levels at capacitance arteries of normotensive rats, and angiotensin II treatment of isolated VSMC abrogated their ability to down-regulate Cks1 and maintain cyclin B protein expression in response to colcemid. Finally, transduction of VSMC from normotensive animals with a retrovirus that drives the expression of Cks1 was sufficient to alter their mitotic spindle cell cycle checkpoint status and promote unscheduled cyclin B metabolism, cell cycle re-entry, and polyploidization. These data demonstrate that Cks1 regulates cyclin B metabolism and ploidy in VSMC and may contribute to the understanding of the phenomena of VSMC polyploidization during hypertension. PMID- 11005811 TI - Mapping the binding site of colchicinoids on beta -tubulin. 2-Chloroacetyl-2 demethylthiocolchicine covalently reacts predominantly with cysteine 239 and secondarily with cysteine 354. AB - 2-Chloroacetyl-2-demethylthiocolchicine (2CTC) and 3-chloroacetyl-3 demethylthiocolchicine (3CTC) resemble colchicine in binding to tubulin and react covalently with beta-tubulin, forming adducts with cysteine residues 239 and 354. The adducts at Cys-239 are less stable than those at Cys-354 during formic acid digestion. Extrapolating to zero time, the Cys-239 to Cys-354 adduct ratio is 77:23 for 2CTC and 27:73 for 3CTC. Using energy minimization modeling to dock colchicinoids into the electron crystallographic model of beta-tubulin in protofilaments (Nogales, E. , Wolf, S. G., and Downing, K. H. (1998) Nature 391, 199-203), we found two potential binding sites. At one, entirely encompassed within beta-tubulin, the C2- and C3-oxygen atoms of 2CTC and 3CTC overlapped poorly with those of colchicine and thiocolchicine, but distances from the reactive carbon atoms of the analogs to the sulfur atoms of the cysteine residues were qualitatively consistent with reactivity. The other potential binding site was located at the alpha/beta interface. Here, the oxygen atoms of the analogs overlapped well with those of colchicine, but relative distances of the reactive carbons to the cysteine sulfur atoms did not correlate with the observed reactivity. A significant conformational change must occur in the colchicine binding site of tubulin in the transition from the unpolymerized to the polymerized state. PMID- 11005812 TI - Multiple amino acid substitutions allow DNA polymerases to synthesize RNA. AB - DNA and RNA polymerase exhibit similarities in structures and catalytic mechanisms, suggesting that both classes of enzymes are evolutionarily related. To probe the biochemical and structure-function relationship between the two classes of polymerases, a large library (200,000 members) of mutant Thermus aquaticus DNA polymerase I (Taq pol I) was created containing random substitutions within a portion of the dNTP binding site (motif A; amino acids 605 617), and a fraction of all selected active Taq pol I (291 of 8000) was tested for the ability to incorporate successive ribonucleotides; 23 unique mutants that added rNTPs into a growing polynucleotide chain were identified and sequenced. These mutants, each containing one to four substitutions, incorporate ribonucleotides at a efficiency approaching 10(3)-fold greater than that of wild type Taq pol I. Several mutants added successive ribonucleotides and thus can catalyze the synthesis of RNA. Sequence analysis of these mutants demonstrates that at least two amino acid residues are involved in excluding ribonucleotides from the active site. Interestingly, wild type DNA polymerases from several distinct families selectively discriminate against rUTP. This study suggests that current DNA and RNA polymerases could have evolved by divergent evolution from an ancestor that shared a common mechanism for polynucleotide synthesis. PMID- 11005813 TI - Mutational analysis of the Pyrococcus furiosus holliday junction resolvase hjc revealed functionally important residues for dimer formation, junction DNA binding, and cleavage activities. AB - The Holliday junction cleavage protein, Hjc resolvase of Pyrococcus furiosus, is the first Holliday junction resolvase to be discovered in Archaea. Although the archaeal resolvase shares certain biochemical properties with other non-archaeal junction resolvases, no amino acid sequence similarity has been identified. To investigate the structure-function relationship of this new Holliday junction resolvase, we constructed a series of mutant hjc genes using site-directed mutagenesis targeted at the residues conserved among the archaeal orthologs. The products of these mutant genes were purified to homogeneity. With analysis of the activity of the mutant proteins to bind and cleave synthetic Holliday junctions, one acidic residue, Glu-9, and two basic residues, Arg-10 and Arg-25, were found to play critical roles in enzyme action. This is in addition to the three conserved residues, Asp-33, Glu-46, and Lys-48, which are also conserved in the motif found in the type II restriction endonuclease family proteins. Two aromatic residues, Phe-68 and Phe-72, are important for the formation of the homodimer probably through hydrophobic interactions. The results of these studies have provided insights into the structure-function relationships of the archaeal Holliday junction resolvase as well as the universality and diversity of the Holliday junction cleavage reaction. PMID- 11005814 TI - Characterization of proteoglycans of human placenta and identification of unique chondroitin sulfate proteoglycans of the intervillous spaces that mediate the adherence of Plasmodium falciparum-infected erythrocytes to the placenta. AB - In pregnant women infected with Plasmodium falciparum, the infected red blood cells (IRBCs) selectively accumulate in the intervillous spaces of placenta, leading to poor fetal outcome and severe health complications in the mother. Although chondroitin 4-sulfate is known to mediate IRBC adherence to placenta, the natural receptor has not been identified. In the present study, the chondroitin sulfate proteoglycans (CSPGs) of human placenta were purified and structurally characterized, and adherence of IRBCs to these CSPGs investigated. The data indicate that the placenta contains three distinct types of CSPGs: significant quantities of uniquely low sulfated, extracellular CSPGs localized in the intervillous spaces, minor amounts of two cell-associated CSPGs, and major amounts of dermatan sulfate-like CSPGs of the fibrous tissue. Of the various CSPGs isolated from the placenta, the low sulfated CSPGs of the intervillous spaces most efficiently bind IRBCs. Based on IRBC adherence capacities and localization patterns of various CSPGs, we conclude that the CSPGs of the intervillous spaces are the receptors for placental IRBC adherence. The identification and characterization of these CSPGs provide a valuable tool for understanding the precise molecular interactions involved in placental IRBC adherence and for the development of therapeutic strategies for maternal malaria. In the accompanying paper (Alkhalil, A., Achur, R. N., Valiyaveettil, M., Ockenhouse, C. F., and Gowda, D. C. (2000) J. Biol. Chem. 275, 40357-40364), we report the structural requirements for the IRBC adherence. PMID- 11005815 TI - Structural requirements for the adherence of Plasmodium falciparum-infected erythrocytes to chondroitin sulfate proteoglycans of human placenta. AB - Plasmodium falciparum infection during pregnancy results in the accumulation of infected red blood cells (IRBCs) in the placenta, leading to poor pregnancy outcome. In the preceding paper (Achur, R. N., Valiyaveettil, M., Alkhalil, A., Ockenhouse, C. F., and Gowda, D. C. (2000) J. Biol. Chem. 275, 40344-40356), we reported that unusually low sulfated chondroitin sulfate proteoglycans (CSPGs) in the intervillous spaces of the placenta mediate the IRBC adherence. In this study, we report the structural requirements for the adherence and the minimum chondroitin 4-sulfate (C4S) structural motif that supports IRBC adherence. Partially sulfated C4Ss with varying sulfate contents were prepared by solvolytic desulfation of a fully sulfated C4S. These and other nonmodified C4Ss, with different proportions of 4-, 6-, and nonsulfated disaccharide repeats, were analyzed for inhibition of IRBC adherence to the placental CSPG. C4Ss containing 30-50% 4-sulfated and 50-70% nonsulfated disaccharide repeats efficiently inhibited IRBC adherence; C6S had no inhibitory activity. Oligosaccharides of varying sizes were prepared by the partial depolymerization of C4Ss containing varying levels of 4-sulfation, and their ability to inhibit the IRBC adherence was studied. Oligosaccharides with six or more disaccharide repeats inhibited IRBC adherence to the same level as that of the intact C4Ss, indicating that a dodecasaccharide is the minimum structural motif required for optimal IRBC adherence. Of the C4S dodecasaccharides, only those with two or three sulfate groups per molecule showed maximum IRBC inhibition. These data define the structural requirements for the IRBC adherence to placental CSPGs with implications for the development of therapeutics for maternal malaria. PMID- 11005816 TI - Aminooxypentane addition to the chemokine macrophage inflammatory protein-1alpha P increases receptor affinities and HIV inhibition. AB - To enter its target cells, human immunodeficiency virus (HIV) must interact with CD4 and one of a family of chemokine receptors. CCR5 is widely used by the virus in this context, and its ligands can prevent HIV entry. Amino-terminal modified chemokine variants, in particular AOP-RANTES (aminooxypentane-linked regulated on activation normal T cell expressed and secreted), exhibit enhanced HIV entry inhibition. We have previously demonstrated that a non-allelic isoform of macrophage inflammatory protein (MIP)-1alpha, termed MIP-1alphaP, is the most active naturally occurring inhibitor of HIV entry known. Here we report the properties of a variant of MIP-1alphaP with an AOP group on the amino terminus. We show that, like RANTES, the addition of AOP to MIP-1alphaP enhances its interactions with CCR1 and CCR5, allows more effective internalization of CCR5, and increases the ligand's potency as an inhibitor of HIV entry through CCR5. Importantly, AOP-MIP-1alphaP is about 10-fold more active than AOP-RANTES at inhibiting HIV entry, making it the most effective chemokine-based inhibitor of HIV entry through CCR5 described to date. Surprisingly, the enhanced receptor interactions of AOP-MIP-1alphaP do not translate into increased chemotaxis or coupling to calcium ion fluxes, suggesting that this protein should be viewed as a partial, rather than a full, agonist for CCR1 and CCR5. PMID- 11005817 TI - Glucocorticoid receptor interaction with 14-3-3 and Raf-1, a proposed mechanism for cross-talk of two signal transduction pathways. AB - The glucocorticoid receptor (GR) functions as a ligand-dependent transcription factor. In the present study we describe a specific immunoaffinity chromatography purification of GR from liver cytosol from adrenalectomized rats that may be used to identify hitherto unknown cytosolic GR interacting proteins. We have identified the ubiquitously expressed 14-3-3 as well as Raf-1, a downstream effector of Ras, as GR co-purifying proteins. In our semi-quantitative analysis liganded/activated GR showed the strongest interaction with 14-3-3 and Raf-1, but 14-3-3 was also found to co-purify with GR in a nonliganded/nonactivated state. By extensive salt washes we were also able to demonstrate that the glucocorticoid induced interaction between GR, 14-3-3, and Raf-1, respectively, is remarkably stable and withstood 2.4 m salt. The interaction between GR and 14-3-3 was also verified by 14-3-3 co-immunoprecipitation studies. Our observations that GR and Raf-1 are found within the same protein complex ("receptosome") in the cytoplasm of rat liver cells could provide a mechanistic explanation for glucocorticoid effects on the Raf-1-Ras signaling pathway. PMID- 11005819 TI - Gene conversion (recombination) mediates expansions of CTG[middle dot]CAG repeats. AB - Genetic recombination is a robust mechanism for expanding CTG.CAG triplet repeats involved in the etiology of hereditary neurological diseases (Jakupciak, J. P., and Wells, R. D. (1999) J. Biol. Chem. 274, 23468-23479). This two-plasmid recombination system in Escherichia coli with derivatives of pUC19 and pACYC184 was used to investigate the effect of triplet repeat orientation on recombination and extent of expansions; tracts of 36, 50, 80, and 36, 100, and 175 repeats in length, respectively, in all possible permutations of length and in both orientations (relative to the unidirectional replication origins) revealed little or no effect of orientation of expansions. The extent of expansions was generally severalfold the length of the progenitor tract and frequently exceeded the combined length of the two tracts in the cotransformed plasmids. Expansions were much more frequent than deletions. Repeat tracts bearing two G-to-A interruptions (polymorphisms) within either 171- or 219-base pair tracts substantially reduced the expansions compared with uninterrupted repeat tracts of similar lengths. Gene conversion, rather than crossing over, was the recombination mechanism. Prior studies showed that DNA replication, repair, and tandem duplication also mediated genetic instabilities of the triplet repeat sequence. However, gene conversion (recombinational repair) is by far the most powerful expansion mechanism. Thus, we propose that gene conversion is the likely expansion mechanism for myotonic dystrophy, spinocerebellar ataxia type 8, and fragile X syndrome. PMID- 11005818 TI - Evidence for a two-electron transfer using the all-ferrous Fe protein during nitrogenase catalysis. AB - The nitrogenase-catalyzed H(2) evolution and acetylene-reduction reactions using Ti(III) and dithionite (DT) as reductants were examined and compared under a variety of conditions. Ti(III) is known to make the all-ferrous Fe protein ([Fe(4)S(4)](0)) and lowers the amount of ATP hydrolyzed during nitrogenase catalysis by approximately 2-fold. Here we further investigate this behavior and present results consistent with the Fe protein in the [Fe(4)S(4)](0) redox state transferring two electrons ([Fe(4)S(4)](2+)/[Fe(4)S(4)](0)) per MoFe protein interaction using Ti(III) but transferring only one electron ([Fe(4)S(4)](2+)/[Fe(4)S(4)](1+)) using DT. MoFe protein specific activity was measured as a function of Fe:MoFe protein ratio for both a one- and a two electron transfer reaction, and nearly identical curves were obtained. However, Fe protein specific activity curves as a function of MoFe:Fe protein ratio showed two distinct reactivity patterns. With DT as reductant, typical MoFe inhibition curves were obtained for operation of the [Fe(4)S(4)](2+)/[Fe(4)S(4)](1+) redox couple, but with Ti(III) as reductant the [Fe(4)S(4)](2+)/[Fe(4)S(4)](0) redox couple was functional and MoFe inhibition was not observed at high MoFe:Fe protein ratios. With Ti(III) as reductant, nitrogenase catalysis produced hyperbolic curves, yielding a V(max) for the Fe protein specific activity of about 3200 nmol of H(2) min(-1) mg(-1) Fe protein, significantly higher than for reactions conducted with DT as reductant. Lag phase experiments (Hageman, R. V., and Burris, R. H. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 2699-2702) were carried out at MoFe:Fe protein ratios of 100 and 300 using both DT and Ti(III). A lag phase was observed for DT but, with Ti(III) product formation, began immediately and remained linear for over 30 min. Activity measurements using Av Cp heterologous crosses were examined using both DT and Ti(III) as reductants to compare the reactivity of the [Fe(4)S(4)](2+)/[Fe(4)S(4)](1+) and [Fe(4)S(4)](2+)/[Fe(4)S(4)](0) redox couples and both were inactive. The results are discussed in terms of the Fe protein transferring two electrons per MoFe protein encounter using the [Fe(4)S(4)](2+)/[Fe(4)S(4)](0) redox couple with Ti(III) as reductant. PMID- 11005820 TI - Determinants for calmodulin binding on voltage-dependent Ca2+ channels. AB - Calmodulin, bound to the alpha(1) subunit of the cardiac L-type calcium channel, is required for calcium-dependent inactivation of this channel. Several laboratories have suggested that the site of interaction of calmodulin with the channel is an IQ-like motif in the carboxyl-terminal region of the alpha(1) subunit. Mutations in this IQ motif are linked to L-type Ca(2+) current (I(Ca)) facilitation and inactivation. IQ peptides from L, P/Q, N, and R channels all bind Ca(2+)calmodulin but not Ca(2+)-free calmodulin. Another peptide representing a carboxyl-terminal sequence found only in L-type channels (designated the CB domain) binds Ca(2+)calmodulin and enhances Ca(2+)-dependent I(Ca) facilitation in cardiac myocytes, suggesting the CB domain is functionally important. Calmodulin blocks the binding of an antibody specific for the CB sequence to the skeletal muscle L-type Ca(2+) channel, suggesting that this is a calmodulin binding site on the intact protein. The binding of the IQ and CB peptides to calmodulin appears to be competitive, signifying that the two sequences represent either independent or alternative binding sites for calmodulin rather than both sequences contributing to a single binding site. PMID- 11005821 TI - SUMO-1 modification of bovine papillomavirus E1 protein is required for intranuclear accumulation. AB - The E1 protein is a multifunctional, origin-binding helicase that is essential for replication of papillomaviruses. Recently, bovine papillomavirus E1 was shown to be post-translationally modified by the addition of the SUMO-1 polypeptide. Here we show that the site of sumoylation maps to lysine residue 514. This lysine and the flanking sequences are well conserved in human papillomavirus (HPV) E1 proteins. Both HPV1a and HPV18 E1 proteins are substrates for sumoylation in vitro, which is consistent with this modification being a general property of E1 proteins. Mutations, which impair the sumoylation of bovine papillomavirus E1, prevent normal nuclear accumulation of E1 with a concomitant loss of replication capacity. These results suggest that sumoylation plays a role in nuclear transport and could regulate the E1 replication function by controlling access to the nuclear replication domains. PMID- 11005822 TI - The putative coiled coil domain of the phi 29 terminal protein is a major determinant involved in recognition of the origin of replication. AB - The linear double-stranded genome of phage phi29 contains a terminal protein (TP) covalently linked at each 5' DNA end, called parental TP. Initiation of phi29 DNA replication starts with the recognition of the origins of replication, constituted by the parental TP-containing DNA ends, by a heterodimer containing phi29 DNA polymerase and primer TP. It has been argued that origin recognition involves protein-protein interactions between parental and primer TP. Analysis of the TP sequence revealed that the region between amino acids 84 and 118 has a high probability to form an amphipatic alpha-helix that could be involved in the interaction between parental and primer TP. Therefore, this TP region may be important for origin recognition. To test this hypothesis we introduced various mutations in the predicted amphipatic alpha-helix and analyzed the functionality of the corresponding purified TP mutants. The results obtained show that the identified putative amphipatic alpha-helix of TP is an important determinant involved in origin recognition. PMID- 11005823 TI - Copper- and zinc-containing superoxide dismutase can act as a superoxide reductase and a superoxide oxidase. AB - The copper- and zinc-containing superoxide dismutase can catalyze the oxidation of ferrocyanide by O(2) as well as the reduction of ferricyanide by O(2). Thus, it can act as a superoxide dismutase (SOD), a superoxide reductase (SOR), and a superoxide oxidase (SOO). The human manganese-containing SOD does not exert SOR or SOO activities with ferrocyanide or ferricyanide as the redox partners. It is possible that some biological reductants can take the place of ferrocyanide and can also interact with human manganese-containing superoxide dismutase, thus making the SOR activity a reality for both SODs. The consequences of this possibility vis a vis H(2)O(2) production, the overproduction of SODs, and the role of copper- and zinc-containing superoxide dismutase mutations in causing familial amyotrophic lateral sclerosis are discussed, as well as the likelihood that the biologically effective SOD mimics, as described to date, actually function as SORs. PMID- 11005824 TI - Molecular modeling and site-directed mutagenesis define the catalytic motif in human gamma -glutamyl hydrolase. AB - Human gamma-glutamyl hydrolase (hGH) is a central enzyme in folyl and antifolylpoly-gamma-glutamate metabolism, which functions by catalyzing the cleavage of the gamma-glutamyl chain of substrates. We previously reported that Cys-110 is essential for activity. Using the sequence of hGH as a query, alignment searches of protein data bases were made using the SSearch and TPROBE programs. Significant similarity was found between hGH and the glutamine amidotransferase type I domain of Escherichia coli carbamoyl phosphate synthetase. The resulting hypothesis is that the catalytic fold of hGH is similar to the folding of this domain in carbamoyl phosphate synthetase. This model predicts that Cys-110 of hGH is the active site nucleophile and forms a catalytic triad with residues His-220 and Glu-222. The hGH mutants C110A, H220A, and E222A were prepared. Consistent with the model, mutants C110A and H220A were inactive. However, the V(max) of the E222A hGH mutant was reduced only 6-fold relative to the wild-type enzyme. The model also predicted that His-171 in hGH may be involved in substrate binding. The H171N hGH mutant was found to have a 250-fold reduced V(max). These studies to determine the catalytic mechanism begin to define the three dimensional interactions of hGH with poly-gamma-glutamate substrates. PMID- 11005825 TI - Evidence for sequential action of cdc7 and cdk2 protein kinases during initiation of DNA replication in Xenopus egg extracts. AB - To investigate how the protein kinase cdc7 stimulates DNA replication in metazoans, a soluble cell-free replication system derived from Xenopus eggs was used. DNA was incubated in egg cytosol to form prereplication complexes and then in nucleoplasmic extract to initiate DNA synthesis. We find that cdc7 is greatly enriched in nucleoplasmic extract and that this high concentration is essential for efficient DNA replication, supporting previous models that the nucleus activates replication indirectly by sequestering essential components. cdc7 binds to chromatin at the G(1)/S transition before initiation occurs, and it dissociates from chromatin as S phase progresses. The chromatin association of cdc7 requires chromatin-bound MCM. In turn, cdc7 is required to load the initiation factor cdc45 onto the DNA. Finally, efficient replication is observed when chromatin is exposed first to cdc7 and then to cdk2 but not when it is exposed to cdk2 before cdc7. Therefore, the cdc7- and cdk2-dependent initiation steps can be separated, indicating the existence of a novel, stable initiation intermediate. Moreover, the data suggest that cdk2 can only act after cdc7 has executed its function. PMID- 11005826 TI - Structural basis of the drastically increased initial electron transfer rate in the reaction center from a Rhodopseudomonas viridis mutant described at 2.00-A resolution. AB - It has previously been shown that replacement of the residue His L168 with Phe (HL168F) in the Rhodopseudomonas viridis reaction center (RC) leads to an unprecedented drastic acceleration of the initial electron transfer rate. Here we describe the determination of the x-ray crystal structure at 2.00-A resolution of the HL168F RC. The electron density maps confirm that a hydrogen bond from the protein to the special pair is removed by this mutation. Compared with the wild type RC, the acceptor of this hydrogen bond, the ring I acetyl group of the "special pair" bacteriochlorophyll, D(L), is rotated, and its acetyl oxygen is found 1.1 A closer to the bacteriochlorophyll-Mg(2+) of the other special pair bacteriochlorophyll, D(M). The rotation of this acetyl group and the increased interaction between the D(L) ring I acetyl oxygen and the D(M)-Mg(2+) provide the structural basis for the previously observed 80-mV decrease in the D(+)/D redox potential and the drastically increased rate of initial electron transfer to the accessory bacteriochlorophyll, B(A). The high quality of the electron density maps also allowed a reliable discussion of the mode of binding of the triazine herbicide terbutryn at the binding site of the secondary quinone, Q(B). PMID- 11005827 TI - Counting probability distributions: differential geometry and model selection. AB - A central problem in science is deciding among competing explanations of data containing random errors. We argue that assessing the "complexity" of explanations is essential to a theoretically well-founded model selection procedure. We formulate model complexity in terms of the geometry of the space of probability distributions. Geometric complexity provides a clear intuitive understanding of several extant notions of model complexity. This approach allows us to reconceptualize the model selection problem as one of counting explanations that lie close to the "truth." We demonstrate the usefulness of the approach by applying it to the recovery of models in psychophysics. PMID- 11005828 TI - Cooperative regulation of light-harvesting complex II phosphorylation via the plastoquinol and ferredoxin-thioredoxin system in chloroplasts. AB - Light induces phosphorylation of photosystem II (PSII) proteins in chloroplasts by activating the protein kinase(s) via reduction of plastoquinone and the cytochrome b(6)f complex. The recent finding of high-light-induced inactivation of the phosphorylation of chlorophyll a/b-binding proteins (LHCII) of the PSII antenna in floated leaf discs, but not in vitro, disclosed a second regulatory mechanism for LHCII phosphorylation. Here we show that this regulation of LHCII phosphorylation is likely to be mediated by the chloroplast ferredoxin thioredoxin system. We present a cooperative model for the function of the two regulation mechanisms that determine the phosphorylation level of the LHCII proteins in vivo, based on the following results: (i) Chloroplast thioredoxins f and m efficiently inhibit LHCII phosphorylation. (ii) A disulfide bond in the LHCII kinase, rather than in its substrate, may be a target component regulated by thioredoxin. (iii) The target disulfide bond in inactive LHCII kinase from dark-adapted leaves is exposed and easily reduced by external thiol mediators, whereas in the activated LHCII kinase the regulatory disulfide bond is hidden. This finding suggests that the activation of the kinase induces a conformational change in the enzyme. The active state of LHCII kinase prevails in chloroplasts under low-light conditions, inducing maximal phosphorylation of LHCII proteins in vivo. (iv) Upon high-light illumination of leaves, the target disulfide bond becomes exposed and thus is made available for reduction by thioredoxin, resulting in a stable inactivation of LHCII kinase. PMID- 11005829 TI - Genes identified by an expression screen of the vector mosquito Anopheles gambiae display differential molecular immune response to malaria parasites and bacteria. AB - We performed a gene expression screen of the entire transcriptome of the major African malaria vector Anopheles gambiae for immune response genes in adult female mosquitoes, which is the developmental stage infected by malaria parasites. Mosquitoes were immune-stimulated for subtractive cloning by treatment with bacterial lipopolysaccharide, a potent and general elicitor of the innate immune response, and by injury. The screen yielded a highly enriched cDNA library in which more than half of the clones were immune responsive. In this paper, we describe 23 immune-regulated genes, including putative protease inhibitors, serine proteases, regulatory molecules, and a number of genes without known relatives. A molecule related to the protease inhibitor alpha-2-macroglobulin responded strongly to malaria parasite infection, but displayed little or no response to bacteria, whereas other genes exhibited the inverse pattern. These results indicate that the insect immune system discriminates between molecular signals specific to infection with bacteria and malaria parasites. PMID- 11005830 TI - A biosensor assay for studying ligand-membrane receptor interactions: binding of antibodies and HIV-1 Env to chemokine receptors. AB - The HIV envelope (Env) protein mediates entry into cells by binding CD4 and an appropriate coreceptor, which triggers structural changes in Env that lead to fusion between the viral and cellular membranes. The major HIV-1 coreceptors are the seven transmembrane domain chemokine receptors CCR5 and CXCR4. The type of coreceptor used by a virus strain is an important determinant of viral tropism and pathogenesis, and virus-receptor interactions can be therapeutic targets. However, Envs from many virus strains interact with CXCR4 and CCR5 with low affinity such that direct study of this important interaction is difficult if not impossible using standard cell-surface binding techniques. We have developed an approach that makes it possible to study ligand binding to membrane proteins, including Env-coreceptor interactions, using an optical biosensor. CCR5, CXCR4, and other membrane proteins were incorporated into retrovirus particles, which were purified and attached to the biosensor surface. Binding of conformationally sensitive antibodies as well as Env to these receptors was readily detected. The equilibrium dissociation constant for the interaction between an Env derived from the prototype HIV-1 strain IIIB for CXCR4 was approximately 500 nM, explaining the difficulty in measuring this interaction using standard equilibrium binding techniques. Retroviral pseudotypes represent easily produced, stable, homogenous structures that can be used to present a wide array of single and multiple membrane-spanning proteins in a native lipid environment for biosensor studies, thus avoiding the need for detergent solubilization, purification, and reconstitution. The approach should have general applicability and can be used to correlate Env-receptor binding constants to viral tropism and pathogenesis. PMID- 11005832 TI - Proliferation is necessary for both repair and mutation in transgenic mouse cells. AB - Proliferating cells are often presumed to be more mutable than quiescent cells because they have less time to repair DNA damage before DNA replication. Direct tests of this hypothesis have been confounded by the need for cell division before a mutation can be detected. We have avoided this problem by showing that the Big Blue mouse cell line permits the dynamic quantification of both lesions and mutations in the complete absence of cell division. These cells carry the bacterial lacI gene in a lambda shuttle vector. Mutant plaques recovered by in vitro packaging of the mouse DNA can arise from mutations sustained either in mouse cells or in the bacteria. The proportion of mutant phage contained within a mutant plaque can distinguish these two types of mutation. Mutations formed in mouse cells yield >90% mutant phage because both DNA strands are mutant. On the other hand, mutations formed in the bacteria from adducted DNA yield 50 copies/ml and virus env gene sequence changes. Each had received a suboptimal regimen before starting HAART. Antiretroviral-resistant HIV-1 can be selected from residual virus replication during HAART in the absence of sustained rebound of plasma HIV-1 RNA. PMID- 11005868 TI - Microglial activation precedes acute neurodegeneration in Sandhoff disease and is suppressed by bone marrow transplantation. AB - Sandhoff disease is a lysosomal storage disorder characterized by the absence of beta-hexosaminidase and storage of G(M2) ganglioside and related glycolipids in the central nervous system. The glycolipid storage causes severe neurodegeneration through a poorly understood pathogenic mechanism. In symptomatic Sandhoff disease mice, apoptotic neuronal cell death was prominent in the caudal regions of the brain. cDNA microarray analysis to monitor gene expression during neuronal cell death revealed an upregulation of genes related to an inflammatory process dominated by activated microglia. Activated microglial expansion, based on gene expression and histologic analysis, was found to precede massive neuronal death. Extensive microglia activation also was detected in a human case of Sandhoff disease. Bone marrow transplantation of Sandhoff disease mice suppressed both the explosive expansion of activated microglia and the neuronal cell death without detectable decreases in neuronal G(M2) ganglioside storage. These results suggest a mechanism of neurodegeneration that includes a vigorous inflammatory response as an important component. Thus, this lysosomal storage disease has parallels to other neurodegenerative disorders, such as Alzheimer's and prion diseases, where inflammatory processes are believed to participate directly in neuronal cell death. PMID- 11005869 TI - Enrichment for murine keratinocyte stem cells based on cell surface phenotype. AB - The identification and physical isolation of epithelial stem cells is critical to our understanding of their growth regulation during homeostasis, wound healing, and carcinogenesis. These stem cells remain poorly characterized because of the absence of specific molecular markers that permit us to distinguish them from their progeny, the transit amplifying (TA) cells, which have a more restricted proliferative potential. Cell kinetic analyses have permitted the identification of murine keratinocyte stem cells (KSCs) as slowly cycling cells that retain [(3)H]thymidine ([(3)H]Tdr) label, termed label-retaining cells (LRCs), whereas TA cells are visualized as rapidly cycling cells after a single pulse of [(3)H]Tdr, termed pulse-labeled cells (PLCs). Here, we report on the successful separation of KSCs from TA cells through the combined use of in vivo cell kinetic analysis and fluorescence-activated cell sorting. Specifically, we demonstrate that murine dorsal keratinocytes characterized by their high levels of alpha(6) integrin and low to undetectable expression of the transferrin receptor (CD71) termed alpha(6)(bri)CD71(dim) cells, are enriched for epithelial stem cells because they represent a minor ( approximately 8%) and quiescent subpopulation of small blast-like cells, with a high nuclear:cytoplasmic ratio, containing approximately 70% of label-retaining cells, the latter being a well documented characteristic of stem cells. Conversely, TA cells could be enriched in a phenotypically distinct subpopulation termed alpha(6)(bri)CD71(bri), representing the majority ( approximately 60%) of basal keratinocytes that are actively cycling, and importantly contain approximately 70% of [(3)H]Tdr pulse-labeled cells. Importantly, immunostaining of dorsal skin revealed the presence of CD71(dim) cells in the hair follicle bulge region, a well documented location for KSCs. PMID- 11005870 TI - Rapid induction of senescence in human cervical carcinoma cells. AB - Expression of the bovine papillomavirus E2 regulatory protein in human cervical carcinoma cell lines repressed expression of the resident human papillomavirus E6 and E7 oncogenes and within a few days caused essentially all of the cells to synchronously display numerous phenotypic markers characteristic of cells undergoing replicative senescence. This process was accompanied by marked but in some cases transient alterations in the expression of cell cycle regulatory proteins and by decreased telomerase activity. We propose that the human papillomavirus E6 and E7 proteins actively prevent senescence from occurring in cervical carcinoma cells, and that once viral oncogene expression is extinguished, the senescence program is rapidly executed. Activation of endogenous senescence pathways in cancer cells may represent an alternative approach to treat human cancers. PMID- 11005871 TI - The essential protein encoded by the UL31 gene of herpes simplex virus 1 depends for its stability on the presence of UL34 protein. AB - To pursue an earlier observation that the protein encoded by the U(L)34 gene binds to intermediate chain of dynein, we constructed a series of mutants from which sequences encoding the entire protein (DeltaU(L)34) or amino-terminal [U(L)34Delta(3-119)] or carboxyl-terminal [U(L)34Delta(245-275)] domains were deleted. The mutant lacking the sequence encoding the carboxyl-terminal domain grew in all cell lines tested. The two other mutants replicated only in cell type dependent manner and poorly. Rescue of DeltaU(L)34 mutant with a fragment that does not encompass the U(L)31 ORF restored wild-type phenotype. U(L)34 protein interacts physically with U(L)31, and the U(L)31 deletion mutant appears to have a phenotype similar to that of U(L)34 deletion mutant. Experiments designed to determine whether the phenotypes of the deletion mutants have a common base revealed that cells infected with the DeltaU(L)34 mutant accumulate U(L)31 RNA but not the corresponding protein. The U(L)31 protein accumulated, however, to near wild-type virus-infected cell levels in cells infected with DeltaU(L)34 mutant and treated with the MG132 proteosomal inhibitor at 6 h after infection. This is evidence that the stability of an essential viral protein requires the presence of another protein. The observation raises the bar for identification of gene function on the basis of analyses of the phenotype of mutants in which the gene has been deleted or rendered inoperative. PMID- 11005872 TI - Cerebral protein synthesis in a genetic mouse model of phenylketonuria. AB - Local rates of cerebral protein synthesis (lCPS(leu)) were measured with the quantitative autoradiographic [1-(14)C]leucine method in a genetic mouse model (Pah(enu2)) of phenylketonuria. As in the human disease, Pah(enu2) mice have a mutation in the gene for phenylalanine hydroxylase. We compared adult homozygous (HMZ) and heterozygous (HTZ) Pah(enu2) mice with the background strain (BTBR). Arterial plasma concentrations of phenylalanine (Phe) were elevated in both HMZ and HTZ mutants by 21 times and 38%, respectively. In the total acid-soluble pool in brain concentrations of Phe were higher and other neutral amino acids lower in HMZ mice compared with either HTZ or BTBR mice indicating a partial saturation of the l-amino acid carrier at the blood brain barrier by the elevated plasma Phe concentrations. In a series of steady-state experiments, the contribution of leucine from the arterial plasma to the tRNA-bound pool in brain was found to be statistically significantly reduced in HMZ mice compared with the other groups, indicating that a greater fraction of leucine in the precursor pool for protein synthesis is derived from protein degradation. We found reductions in lCPS(leu) of about 20% throughout the brain in the HMZ mice compared with the other two groups, but no reductions in brain concentrations of tRNA-bound neutral amino acids. Our results in the mouse model suggest that in untreated phenylketonuria in adults, the partial saturation of the l-amino acid transporter at the blood brain barrier may not underlie a reduction in cerebral protein synthesis. PMID- 11005873 TI - An approach to probe some neural systems interaction by functional MRI at neural time scale down to milliseconds. AB - In this paper, we demonstrate an approach by which some evoked neuronal events can be probed by functional MRI (fMRI) signal with temporal resolution at the time scale of tens of milliseconds. The approach is based on the close relationship between neuronal electrical events and fMRI signal that is experimentally demonstrated in concurrent fMRI and electroencephalographic (EEG) studies conducted in a rat model with forepaw electrical stimulation. We observed a refractory period of neuronal origin in a two-stimuli paradigm: the first stimulation pulse suppressed the evoked activity in both EEG and fMRI signal responding to the subsequent stimulus for a period of several hundred milliseconds. When there was an apparent site-site interaction detected in the evoked EEG signal induced by two stimuli that were primarily targeted to activate two different sites in the brain, fMRI also displayed signal amplitude modulation because of the interactive event. With visual stimulation using two short pulses in the human brain, a similar refractory phenomenon was observed in activated fMRI signals in the primary visual cortex. In addition, for interstimulus intervals shorter than the known latency time of the evoked potential induced by the first stimulus ( approximately 100 ms) in the primary visual cortex of the human brain, the suppression was not present. Thus, by controlling the temporal relation of input tasks, it is possible to study temporal evolution of certain neural events at the time scale of their evoked electrical activity by noninvasive fMRI methodology. PMID- 11005874 TI - Prenatal stress produces learning deficits associated with an inhibition of neurogenesis in the hippocampus. AB - Early experiences such as prenatal stress significantly influence the development of the brain and the organization of behavior. In particular, prenatal stress impairs memory processes but the mechanism for this effect is not known. Hippocampal granule neurons are generated throughout life and are involved in hippocampal-dependent learning. Here, we report that prenatal stress in rats induced lifespan reduction of neurogenesis in the dentate gyrus and produced impairment in hippocampal-related spatial tasks. Prenatal stress blocked the increase of learning-induced neurogenesis. These data strengthen pathophysiological hypotheses that propose an early neurodevelopmental origin for psychopathological vulnerabilities in aging. PMID- 11005875 TI - Regional and strain-specific gene expression mapping in the adult mouse brain. AB - To determine the genetic causes and molecular mechanisms responsible for neurobehavioral differences in mice, we used highly parallel gene expression profiling to detect genes that are differentially expressed between the 129SvEv and C57BL/6 mouse strains at baseline and in response to seizure. In addition, we identified genes that are differentially expressed in specific brain regions. We found that approximately 1% of expressed genes are differentially expressed between strains in at least one region of the brain and that the gene expression response to seizure is significantly different between the two inbred strains. The results lead to the identification of differences in gene expression that may account for distinct phenotypes in inbred strains and the unique functions of specific brain regions. PMID- 11005877 TI - High phosphorylation efficiency and depression of uncoupled respiration in mitochondria under hypoxia. AB - Mitochondria are confronted with low oxygen levels in the microenvironment within tissues; yet, isolated mitochondria are routinely studied under air-saturated conditions that are effectively hyperoxic, increase oxidative stress, and may impair mitochondrial function. Under hypoxia, on the other hand, respiration and ATP supply are restricted. Under these conditions of oxygen limitation, any compromise in the coupling of oxidative phosphorylation to oxygen consumption could accentuate ATP depletion, leading to metabolic failure. To address this issue, we have developed the approach of oxygen-injection microcalorimetry and ADP-injection respirometry for evaluating mitochondrial function at limiting oxygen supply. Whereas phosphorylation efficiency drops during ADP limitation at high oxygen levels, we show here that oxidative phosphorylation is more efficient at low oxygen than at air saturation, as indicated by higher ratios of ADP flux to total oxygen flux at identical submaximal rates of ATP synthesis. At low oxygen, the proton leak and uncoupled respiration are depressed, thus reducing maintenance energy expenditure. This indicates the importance of low intracellular oxygen levels in avoiding oxidative stress and protecting bioenergetic efficiency. PMID- 11005876 TI - Experimental diabetes in rats causes hippocampal dendritic and synaptic reorganization and increased glucocorticoid reactivity to stress. AB - We report that 9 d of uncontrolled experimental diabetes induced by streptozotocin (STZ) in rats is an endogenous chronic stressor that produces retraction and simplification of apical dendrites of hippocampal CA3 pyramidal neurons, an effect also observed in nondiabetic rats after 21 d of repeated restraint stress or chronic corticosterone (Cort) treatment. Diabetes also induces morphological changes in the presynaptic mossy fiber terminals (MFT) that form excitatory synaptic contacts with the proximal CA3 apical dendrites. One effect, synaptic vesicle depletion, occurs in diabetes as well as after repeated stress and Cort treatment. However, diabetes produced other MFT structural changes that differ qualitatively and quantitatively from other treatments. Furthermore, whereas 7 d of repeated stress was insufficient to produce dendritic or synaptic remodeling in nondiabetic rats, it potentiated both dendritic atrophy and MFT synaptic vesicle depletion in STZ rats. These changes occurred in concert with adrenal hypertrophy and elevated basal Cort release as well as hypersensitivity and defective shutoff of Cort secretion after stress. Thus, as an endogenous stressor, STZ diabetes not only accelerates the effects of exogenous stress to alter hippocampal morphology; it also produces structural changes that overlap only partially with those produced by stress and Cort in the nondiabetic state. PMID- 11005878 TI - Reactivation of encoding-related brain activity during memory retrieval. AB - Neuronal models predict that retrieval of specific event information reactivates brain regions that were active during encoding of this information. Consistent with this prediction, this positron-emission tomography study showed that remembering that visual words had been paired with sounds at encoding activated some of the auditory brain regions that were engaged during encoding. After word sound encoding, activation of auditory brain regions was also observed during visual word recognition when there was no demand to retrieve auditory information. Collectively, these observations suggest that information about the auditory components of multisensory event information is stored in auditory responsive cortex and reactivated at retrieval, in keeping with classical ideas about "redintegration, " that is, the power of part of an encoded stimulus complex to evoke the whole experience. PMID- 11005880 TI - Leadership: Challenges in the changing health care environment. PMID- 11005881 TI - No-spill sippy cups. PMID- 11005879 TI - Memory's echo: vivid remembering reactivates sensory-specific cortex. AB - A fundamental question in human memory is how the brain represents sensory specific information during the process of retrieval. One hypothesis is that regions of sensory cortex are reactivated during retrieval of sensory-specific information (1). Here we report findings from a study in which subjects learned a set of picture and sound items and were then given a recall test during which they vividly remembered the items while imaged by using event-related functional MRI. Regions of visual and auditory cortex were activated differentially during retrieval of pictures and sounds, respectively. Furthermore, the regions activated during the recall test comprised a subset of those activated during a separate perception task in which subjects actually viewed pictures and heard sounds. Regions activated during the recall test were found to be represented more in late than in early visual and auditory cortex. Therefore, results indicate that retrieval of vivid visual and auditory information can be associated with a reactivation of some of the same sensory regions that were activated during perception of those items. PMID- 11005882 TI - Primary health care for Hispanic children of migrant farm workers. AB - Providing primary care to children of culturally diverse populations is a challenge for pediatric nurse practitioners and educators. The challenge is intensified when providing care to Hispanic children who are uprooted because their parent(s) are migrant farm workers. The creation of health-focused academic community partnerships is one unique strategy to improve primary care to these children. One such partnership is the ongoing Migrant Family Health Program in which practitioner nursing students and their faculty members provide primary health care to children who are enrolled in a summer education program for migrant children. PMID- 11005883 TI - An added dimension to the pediatric health maintenance visit: the spiritual history. AB - The initial pediatric health maintenance visit gives the nurse practitioner (NP) an opportunity to gather information about a family that can be used to devise an effective and culturally sensitive plan of care. NPs have a history of exploring spirituality as a segment of a family's culture to help children and their parents cope with an acute or chronic illness. However, spirituality should also be discussed with a family because it may affect their normal, everyday lives. Religious traditions, values, and ethical concerns may provide rich insight into a family's dynamic, which can increase an NP's understanding. The mnemonic B-E-L I-E-F is used to guide NPs through a discussion of spirituality within a pediatric health care maintenance context. This framework, which incorporates an added dimension of spirituality into the health history, will further advance efforts to assist families in fostering optimal childhood growth and development during a routine health care maintenance visit. PMID- 11005885 TI - Oral contraceptive pills: considerations for the adolescent patient. AB - Combined oral contraceptive pills (OCPs) are the most commonly prescribed method of birth control for adolescents. This article presents an overview of OCP pharmacology and summarizes the different types of OCPs. The initial patient evaluation and subsequent care are described, with a focus on management plans specific to adolescents. Emergency contraception, an alternative use of OCPs, is described as well. A thorough knowledge of OCPs and an appreciation of adolescent specific management plans will enhance nurse practitioners' skills in preventing pregnancy in their adolescent patients. PMID- 11005884 TI - School-based support for urban adolescent mothers. AB - INTRODUCTION: The purpose of this research was to examine infant birth weights and high school completion rates for two cohorts of adolescent mothers attending an urban school-based program, the Polly T. McCabe Center. METHOD: A retrospective record review was conducted with two convenience samples of 47 adolescent mothers (study I) aged 16.1 +/- 1.4 years and 60 adolescent mothers (study II) aged 15.9 +/- 1.4 years who were enrolled in the McCabe Center during the 1992-1993 and 1996-1997 academic years. RESULTS: The incidence of low birth weight infants born to students was 4% in study I and 13.6% in study II. High school continuation/completion rates for study I were 79% at 3 years after leaving the program and 80% for study II at 1 year after leaving the program. DISCUSSION: The young mothers and infants appear to have benefitted from the McCabe Center's intervention. Findings of the study suggest that continued implementation of supportive school-based programs similar to the McCabe Center may help prevent lower birth weight infants born to adolescent mothers and may help decrease the high school drop-out rate among this population. PMID- 11005886 TI - Perceptions of children with HIV infection when not told for so long: implications for diagnosis disclosure. AB - INTRODUCTION: The perceptions of school-aged children with HIV infection about their illness were evaluated in light of parental/guardians communication about the child's diagnosis to clarify guidelines for disclosure. METHOD: An innovative, qualitative research design using methods of grounded theory and projective drawings involved 12 children and 13 parents/guardians from a West Coast Pediatric AIDS Clinical Trial treatment center. RESULTS: A specific process of interaction between parents/guardians and children, when not told for so long, emerged as the central phenomenon of the study. Parents/guardians kept the children's HIV infection secret for 2 to 8 years after diagnosis. Regardless of disclosure status, the children's drawings and conversations suggested severe emotional distress, disturbed self-image, and social isolation. Parents and guardians were unaware of their children's concerns. DISCUSSION: Although other circumstances of the children's lives contributed to their poor psychosocial adjustment, the long interval of silence about the illness also played a part in their poor adjustment. Parents/guardians did not recognize when their children needed more supportive information. Pediatric clinicians should therefore continually assess children's psychosocial adjustment to guide families through the disclosure process. The use of projective drawing techniques can facilitate this process. PMID- 11005887 TI - Neurofibromatosis: an elephant by another name. PMID- 11005888 TI - Fluoride supplementation and caries prevention. PMID- 11005889 TI - Case of a school-aged child with a limp and hip pain. PMID- 11005890 TI - Politics set the agenda for children--and for pediatric nurse practitioners. PMID- 11005891 TI - Getting ready for "the big talk" and beyond. PMID- 11005892 TI - Questions & answers. PMID- 11005894 TI - The Swiss Society for Prosthetic Dentistry-founded in 1980. PMID- 11005893 TI - Indian Prosthodontic Society - founded in 1973. PMID- 11005895 TI - Sectional collapsed denture for a partially edentulous patient with microstomia: a clinical report. PMID- 11005896 TI - Trial anterior artificial tooth arrangement for an immediate denture patient: a clinical report. PMID- 11005897 TI - Management of unfavorable implant placement: a clinical report. PMID- 11005898 TI - Prosthodontic management of limited oral access after ablative tumor surgery: a clinical report. PMID- 11005899 TI - Corrosion-fatigue life of commercially pure titanium and Ti-6Al-4V alloys in different storage environments. AB - STATEMENT OF PROBLEM: Removable partial dentures are affected by fatigue because of the cyclic mechanism of the masticatory system and frequent insertion and removal. Titanium and its alloys have been used in the manufacture of denture frameworks; however, preventive agents with fluorides are thought to attack titanium alloy surfaces. PURPOSE: This study evaluated, compared, and analyzed the corrosion-fatigue life of commercially pure titanium and Ti-6Al-4V alloy in different storage environments. MATERIAL AND METHODS: For each metal, 33 dumbbell rods, 2.3 mm in diameter at the central segment, were cast in the Rematitan system. Corrosion-fatigue strength test was carried out through a universal testing machine with a load 30% lower than the 0.2% offset yield strength and a combined influence of different environments: in air at room temperature, with synthetic saliva, and with fluoride synthetic saliva. After failure, the number of cycles were recorded, and fracture surfaces were examined with an SEM. RESULTS: ANOVA and Tukey's multiple comparison test indicated that Ti-6Al-4V alloy achieved 21,269 cycles (SD = 8,355) against 19,157 cycles (SD = 3, 624) for the commercially pure Ti. There were no significant differences between either metal in the corrosion-fatigue life for dry specimens, but when the solutions were present, the fatigue life was significantly reduced, probably because of the production of corrosion pits caused by superficial reactions. PMID- 11005900 TI - Differences in bonding to acid-etched or Er:YAG-laser-treated enamel and dentin surfaces. AB - STATEMENT OF PROBLEM: Er:YAG (erbium-doped yttrium aluminium garnet) lasers have been effective in the removal of dental tissues. It has been suggested that they are also useful for preparing dental surfaces for adhesion, but results to date have been controversial. PURPOSE: This study compared the tensile strength of bracket-tooth bonds obtained after preparation of the surface for adhesion (dentin or enamel) by conventional acid-etching or by Er:YAG laser etching and investigated microstructure of resin-tooth interfaces using the 2 procedures. MATERIAL AND METHODS: Eighty healthy human premolars were used. Brackets were cemented to acid-etched enamel, laser-etched enamel, acid-etched dentin, or laser etched dentin (20 teeth per group). Dentin was previously exposed using a high speed handpiece. Acid-etching was with 37% orthophosphoric acid (15 seconds for enamel, 5 seconds for dentin). Laser etching was with Er:YAG laser (four 200 mJ pulses per second for enamel; four 160 mJ pulses per second for dentin). Brackets were bonded with autocuring resin paste, having first applied a primer (dentin only) and then light-cured bonding resin. Tensile strength was determined with a universal testing machine. Data were analyzed with 2-way ANOVA and subsequent t test with Bonferroni correction. Fracture patterns were compared by the Wilcoxon test with Bonferroni correction. For SEM studies of the resin-tooth interface, a total of 12 premolars were used (3 for each tissue per treatment combination). RESULTS: Mean tensile bond strength for acid-etched enamel (14.05 +/- 5.03 MPa) was significantly higher (P<.05) than for laser-etched enamel (8.45 +/- 3.07 MPa), and significantly higher (P<.05) for acid-etched dentin (4.70 +/- 2.50 MPa) than laser-etched dentin (2.48 +/- 1.94 MPa). Bond failure after laser etching was due to microcohesive fracture of tooth tissue. SEM studies of both resin enamel and resin-dentin interfaces indicated extensive subsurface fissuring after laser etching. CONCLUSION: Adhesion to dental hard tissues after Er:YAG laser etching is inferior to that obtained after conventional acid etching. Enamel and dentin surfaces prepared by Er:YAG laser etching show extensive subsurface fissuring that is unfavorable to adhesion. PMID- 11005901 TI - Three-year clinical evaluation of direct and indirect composite restorations in posterior teeth. AB - STATEMENT OF THE PROBLEM: Objective long-term clinical data are necessary to assess the performance of modern posterior composites as direct and indirect restorations. PURPOSE: This prospective, long-term clinical trial evaluated direct and indirect composite restorations for clinical acceptability as posterior restoratives in single or multisurface cavities and provided a survey on the 3-year results. MATERIAL AND METHODS: Under the supervision of an experienced dentist, 9 dental students placed 88 composite restorations (Tetric, blend-a-lux, Pertac-Hybrid Unifil), 43 direct composite restorations, and 45 indirect inlays. Clinical evaluation was performed at baseline and in yearly intervals after placement by 2 other experienced dentists, using modified USPHS criteria. A third follow-up of 60 restorations took place within 33 to 36 months after placement. RESULTS: A total of 93% of indirect and 87% of direct restorations were assessed to be clinically excellent or acceptable. During the third year, 1 direct restoration in a molar failed because of margin opening. Indirect inlays exhibited a significantly better anatomic form of the surface than direct composite restorations. Premolars revealed a significantly better marginal integrity and anatomic form of the surface than molars. Restorations in molars exhibited a significantly higher failure rate compared with premolars. CONCLUSION: Posterior composite restorations provided a satisfactory clinical performance over a 3-year period, even if placed by relatively inexperienced but supervised students. PMID- 11005902 TI - Fracture resistance of Class II approximal slot restorations. AB - STATEMENT OF PROBLEM: Determination of the fracture resistance of various restorative materials in Class II approximal slot restorations has not been studied. PURPOSE: This study evaluated the effects of retention grooves and different restorative materials in Class II approximal slot restorations. To explore the possibilities for further research, the probable effects of preparation size and loading angle were investigated in a limited manner. MATERIAL AND METHODS: Ninety sound, caries-free human maxillary premolars were divided into 9 groups. The cavities were prepared either by hand or in a computer controlled CNC machine with or without retention grooves. Four were restored with adhesive amalgam, another 4 with composite, and a single group with Compomer resin. The gingival floor depth was 1.5 mm. The specimens were loaded at an angle of 13. 5 degrees to their longitudinal axes by using a computer-controlled material testing machine until failure occurred. For one specific preparation of adhesive amalgam, loading was applied at 0 and 30 degrees to determine the probable effects of the loading angle. For a specific composite, resin application, the effects of the change in gingival floor depth were analyzed by assigning the depth to 2.0 mm. RESULTS: Composite and Compomer resin and composite exhibited better performance than amalgam. The existence of the retention grooves proved to be effective for adhesive amalgam restorations but did not have any advantageous effect in composite and Compomer restoration. CONCLUSION: For improved fracture resistance in small approximal restorations, the use of composite was the appropriate choice. Compomer also gave satisfactory results. Use of amalgam restoration should be accompanied with retention grooves and an adhesive system to improve its performance. PMID- 11005903 TI - Fracture resistance of posterior metal-free polymer crowns. AB - STATEMENT OF PROBLEM: Improved mechanical properties of contemporary composites has resulted in the extended use of composites for the restoration of posterior teeth. Although the indication of polymers was extended to metal-free individual crowns, the influence of tooth preparation design and cementation methods on the stability of these artificial crowns remains unknown. PURPOSE: This in vitro study evaluated the effect of axial tooth preparation design, occlusal dimension, and cementation technique on the fracture resistance of metal-free posterior Artglass crowns. MATERIAL AND METHODS: Seventy-two extracted human third molars, assigned to experimental groups by size, received standardized tooth preparation. Axial tooth preparation included an invasive approach with 1-mm deep shoulder and a less invasive 0.5-mm chamfer preparation, whereas occlusal reduction was either 0.5 mm or 1.3 mm. Artglass crowns that restored the original tooth contour were cemented with 3 cements: zinc phosphate cement (ZnP), glass ionomer cement (GIC), or a resinous cement in combination with a dentinal bonding agent. After 10,000 thermal cycles between 5 degrees C and 55 degrees C, artificial crowns were vertically loaded until compression to failure. Significant differences of fracture loads between experimental groups were assessed by paired Mann-Whitney U tests. RESULTS: Minimal fracture resistance for all combinations excluded 500 N. However, 9 of 24 Artglass crowns cemented with ZnP loosened after thermocycling. Adhesive cementation resulted in a significantly greater fracture resistance compared with GIC and ZnP (P=.02). Increased occlusal thickness (0.5 to 1.3 mm) resulted in greater stability, whereas a 1-mm deep shoulder tooth preparation did not improve durability compared with a 0.5-mm chamfer finishing line. CONCLUSION: A minimally invasive 0.5-mm axial chamfer tooth preparation combined with sufficient occlusal reduction and adhesive cementation recorded the greatest stability for posterior metal-free Artglass crowns. PMID- 11005904 TI - Influence of metal surface finishing on porcelain porosity and beam failure loads at the metal-ceramic interface. AB - STATEMENT OF THE PROBLEM: Investigators suggest that metals should be finished in 1 direction before porcelain application to minimize metal irregularities and trapped contaminants. These irregularities are thought to be focal points for porosity and crack propagation. PURPOSE: This study investigated the influence of metal finishing and sandblasting on (1) porosity production at the porcelain metal interface, and (2) porcelain-metal beam failure load. MATERIAL AND METHODS: Eighty cast metal samples were divided into 4 test groups: (A) bidirectional finish/sandblasting; (B) unidirectional finish/sandblasting (C) bidirectional finish only; and (D) unidirectional finish only. The porcelain applied was 1.5 mm thick. Samples were sectioned longitudinally. Half of the samples were subjected to a 3-point flexural test. The remaining samples were sectioned into 4 slices and were examined with a light microscope (x500). Number and diameter of porosities at the metal-porcelain interface were recorded. RESULTS: Mean loads at failure (lbs) were as follows: A, 11.1 +/- 1.3 (5.03 +/- 0.58 Kg); B, 11.2 +/- 1.7 (5. 08 +/- 0.77 Kg); C, 4.0 +/- 1.8 (1.81 +/- 0.81 Kg); and D, 5.0 +/- 2. 1 (2.26 +/- 0.95 Kg). Groups A and B were significantly different from groups C and D (P<.0001). Nonsandblasted samples (C and D) exhibited a separation at the ceramometal interface, which prevented quantification of porosity size and number. Average interface porosity sizes (microm) (A, 8.99 +/- 1.92; B, 10.03 +/- 1.86) showed no significant difference. The mean interface porosity number (A, 62. 3 +/- 16.02; B, 67.4 +/- 10.01) showed no significant difference. CONCLUSION: Direction of metal finishing did not affect the porosity number and size at the ceramometal interface or the beam failure loads. Sandblasting increased the beam failure loads. Nonsandblasted samples showed detachment of the porcelain from the metal. PMID- 11005905 TI - Experimentally induced abutment strains in three types of single-molar implant restorations. AB - STATEMENT OF PROBLEM: The choice of single-molar implant design is difficult because of a lack of controlled, quantitative biomechanical analyses. PURPOSE: This study determined the effect of 3 single-molar implant designs on implant strains under a variety of homologous loading conditions. MATERIAL AND METHODS: On each implant abutment, 4 strain gauges were placed axially at 90 degrees to each other on the buccal, lingual, mesial, and distal surfaces. Effects of implant design, load location, direction, and magnitude were tested on axial and bending (buccolingual and mesiodistal) strains of 3 single-molar implant designs: (1) single, 3.75-mm (regular) diameter implant, (2) single, 5-mm (wide) diameter implant, and (3) two 3.75-mm diameter (double) implants connected through a single-molar crown. Results were analyzed with ANOVA. RESULTS: Variations in loading conditions induced 3-dimensionally complex abutment strains on the tested implant designs. Peak absolute strains in mesiodistal direction were 6493 microepsilon for design 1 and 3958 microepsilon on design 2, and 3160 microepsilon in buccolingual direction on design 3. For all loading conditions, the single 3.75-mm diameter implant consistently experienced the largest strains compared with wide-diameter and double implant designs. Changes in centric contact location affected implant abutment strains differently among the 3 designs. Angulated force direction resulted in larger bending strains. CONCLUSION: For single-molar implant designs, an increase in implant number and diameter may effectively reduce experimental implant abutment strains. PMID- 11005906 TI - Effect of anchorage systems and extension base contact on load transfer with mandibular implant-retained overdentures. AB - STATEMENT OF PROBLEM: Controversy exists regarding the effect of anchorage systems and extension base contact on stress transfer to multiple implants by mandibular overdentures. PURPOSE: This simulation study measured photoelastically the biologic behavior of 4 implants retaining a mandibular overdenture. The purpose of the investigation was to compare the load transfer characteristics of different mandibular-retained overdenture designs, with and without edentulous ridge contact. MATERIAL AND METHODS: A photoelastic model of a human edentulous mandible was fabricated having 4 screw-type implants (3.75 x 10 mm) embedded in the parasymphyseal area. Substructure designs utilizing a cantilevered bar, spark erosion framework, noncantilevered bar, and solitary anchors were fabricated. A vertical load of 30 lb was applied to the first molar unilaterally on each of the 4 standardized overdenture prostheses, with and without a silicone tissue spacer, for a total of 8 tested conditions. Stresses that developed in the supporting structure were monitored photoelastically and recorded photographically. RESULTS: Without the simulated tissue contact on the posterior edentulous ridge, the cantilevered bar framework caused the highest stresses to the terminal implant, followed by spark erosion framework, non-cantilevered bar, and solitary anchor design. With simulated tissue contact under the extension base, stress transfer to the distal implant was uniformly reduced to a low level. CONCLUSION: Without intimate extension base contact with the posterior edentulous ridge, the cantilevered anchorage systems generated the highest stresses, under load, to the ipsilateral terminal implant and the solitary anchor design transferred the least. With simulated intimate extension base contact, all anchorage systems transferred low stress to the distal implant region. For all conditions and designs, low stress was transferred to the contralateral side of the arch. PMID- 11005908 TI - Custom retainer for the Blom-Singer adjustable tracheostoma valve. AB - This article presents a procedure that allows the laryngectomy patient to be fitted with a well-sealed custom device. The custom device provides near-normal speech during a variety of activities. The custom retainer for the Blom-Singer adjustable tracheostoma valve can be fabricated with medical grade silicone using routine prosthodontic techniques. PMID- 11005907 TI - Effect of adhesive retention on maxillofacial prostheses. Part I: skin dressings and solvent removers. AB - STATEMENT OF PROBLEM: The success of most maxillofacial prostheses depends on retention by medical adhesives. Products such as Smith-Nephew's Skin-Prep (SP) are available that can be used on the skin that could improve prosthesis adhesion protective dressing. The removal of adhesive from the skin is also problematic, so solvents, such as Uni-Solve adhesive remover (US), are often used. PURPOSE: This study measured the removal force of silicone elastomer strips with 2 adhesives from the skin of human subjects during the day, as affected by the use of SP and US, and determined the site of adhesive failure. MATERIAL AND METHODS: Silicone rubber strips were applied in a predetermined random order to the ventral arm surfaces of 20 human subjects. US was applied to half the sites 1 day before testing. SP was also applied to half the sites just before Epithane-3 (E3) or Secure(2) Medical Adhesive (SMA) were used to adhere the strips. They were peeled from the skin 6 hours later in an Instron at a rate of 10 cm/min. RESULTS: A 3-way within-group MANOVA revealed significant differences without interactions between adhesives (SMA=96.3 N.m, E3=24.1 N.m; P<.0005) and between use or nonuse of SP (SP=65.8 N.m, no SP=54.6 N.m; P<.0005). The use of US was not significant (no US=61.8 N.m, with US=58.6 N.m; P=.197). SMA adhered to the prostheses, whereas E3 adhered to the skin, leaving a residue (Fisher exact test; P<.0003). CONCLUSION: The combination of SMA and SP showed the highest adhesive bond strength. Overall, SMA was 3 to 5 times more retentive than E3. SP improved adhesion of both SMA (15%) and E3 (27%). SMA was still far more retentive. US had no effect on retention. SMA remained on the prostheses, whereas E3 left a difficult-to-remove residue on the skin. PMID- 11005909 TI - Use of an ultrahigh-speed laser scanner for constructing three-dimensional shapes of dentition and occlusion. AB - STATEMENT OF PROBLEM: Acquisition of the 3-dimensional shape of dental casts is useful for quantitative evaluation of the diagnosis and treatment of occlusion. PURPOSE: This study demonstrated the acquisition of data through the use of an ultrahigh-speed laser scanner and an originally developed goniometer, the use of these tools to measure the entire 3-dimensional shape of the dental cast from multiple directions, and the connection of the data measurements to reconstruct the cast's occlusion with the use of a computer. MATERIAL AND METHODS: A commercially available apparatus that emits a line laser and completes 1 scan in only 0.6 seconds was used. To compensate for the unmeasured region, the cast was measured from 4 directions on the original goniometer, and the connection of data to obtain the entire image was carried out. Further, the reconstruction of the occlusion between the upper and lower casts was attempted by transferring the upper cast data to the lower cast. RESULTS: By the data connection, the entire shape of the upper and lower casts was constructed, and the characteristic structure was reproduced. The data connections were satisfactory for the flat surfaces but less accurate for the inclined surfaces. Reconstruction of the occlusion between the upper and lower casts was accomplished. It was then possible to visualize the occlusion from arbitrary directions and sections with computer graphics. CONCLUSION: The construction of the entire 3-dimensional shape of a dental cast and the reconstruction of the occlusion were accomplished using an ultrahigh-speed measurement system and original goniometer. These results will be useful for clinical applications such as computerized diagnoses and treatment of occlusion, and for the replacement of the stone casts in the dental office by computerized data. PMID- 11005910 TI - In vivo evaluation of the surface of posterior resin composite restorations: a pilot study. AB - STATEMENT OF PROBLEM: Several methods have been used to determine the surface characteristics of resin composites in vivo and compare composite wear rates with enamel wear rates. PURPOSE: This pilot study describes the surface characteristics of resin composites and the wear of resin composites and enamel during 1 year of in vivo service. MATERIAL AND METHODS: A single Class II posterior resin composite restoration (Z100) was placed in 10 patients. Restored teeth and unrestored adjacent control teeth were measured for wear 4 times within the first year. A null point contact stylus profiler and fitting software were used to measure epoxy casts. Maximum depth of wear, average depth of wear, and characteristics of the restoration margin were determined. Paired t tests were used to compare the control and restored teeth, and ANOVA was used to assess the progression of wear over time (P<.05). RESULTS: After 1 year, maximum depth of wear over the entire preparation region was on average 204.8 microm (+/- 129.8), significantly greater than the 36. 8 microm (+/- 10.1) average maximum depth of wear of enamel at occlusal contact areas on control teeth (P=.009). Maximum depth of wear progressed over time (P=.009). Fracture of excess composite, commonly called flash fracture, occurred in 50% of the restored teeth extending over the preparation margin. CONCLUSION: Composite restorations wore significantly faster than enamel contact areas on control teeth. Also of concern were the marginal flash fractures that could facilitate secondary caries. PMID- 11005911 TI - In vitro and in vivo corrosion evaluation of nickel-chromium- and copper-aluminum based alloys. AB - STATEMENT OF PROBLEM: The low resistance to corrosion is the major problem related to the use of copper-aluminum alloys. PURPOSE: This in vitro and in vivo study evaluated the corrosion of 2 copper-aluminum alloys (Cu-Al and Cu-Al-Zn) compared with a nickel-chromium alloy. MATERIAL AND METHODS: For the in vitro test, specimens were immersed in the following 3 corrosion solutions: artificial saliva, 0.9% sodium chloride, and 1.0% sodium sulfide. For the in vivo test, specimens were embedded in complete dentures, so that one surface was left exposed. The 3 testing sites were (1) close to the oral mucosa (partial self cleaning site), (2) surface exposed to the oral cavity (self-cleaning site), and (3) specimen bottom surface exposed to the saliva by means of a tunnel-shaped perforation (non-self-cleaning site). RESULTS: Almost no corrosion occurred with the nickel-chromium alloy, for either the in vitro or in vivo test. On the other hand, the 2 copper-aluminum-based alloys exhibited high corrosion in the sulfide solution. These same alloys also underwent high corrosion in non-self-cleaning sites for the in vivo test, although minimal attack was observed in self-cleaning sites. CONCLUSION: The nickel-chromium alloy presented high resistance to corrosion. Both copper-aluminum alloys showed considerable corrosion in the sulfide solution and clinically in the non-self-cleaning site. However, in self cleaning sites these 2 alloys did not show substantial corrosion. PMID- 11005912 TI - Fabrication of an indirect-direct provisional fixed partial denture. AB - This article describes a procedure for fabricating an optimal provisional restoration before tooth preparation. The procedure involves the fabrication of an indirect-direct provisional fixed partial denture. There are several advantages to this procedure, such as reduced chair time, less heat generated in the mouth, reduced contact between resin monomer and soft tissues, and optimal esthetics. PMID- 11005913 TI - Overdenture abutments for fixed partial dentures. AB - Despite universal acceptance for retaining endodontically treated and filled roots for vertical support of removable prostheses, there is little evidence for the use of overdenture abutments with fixed prostheses. This article presents a review of clinical situations for using these teeth to support fixed partial dentures. These teeth provide vertical support during function and preserve alveolar bone levels. PMID- 11005914 TI - Simplified duplicating procedure for dental casts. PMID- 11005915 TI - Simple technique for making custom impression trays. PMID- 11005916 TI - Treatment outcomes of fixed or removable implant-supported prostheses in the edentulous maxilla. PMID- 11005917 TI - Reply PMID- 11005918 TI - Transglycosylation by Streptococcus mutans GS-5 glucosyltransferase-D: acceptor specificity and engineering of reaction conditions. AB - The acceptor specificity of Streptococcus mutans GS-5 glucosyltransferase-D (GTF D) was studied, particular the specificity toward non-saccharide compounds. Dihydroxy aromatic compounds like catechol, 4-methylcatechol, and 3 methoxycatechol were glycosylated by GTF-D with a high efficiency. Transglycosylation yields were 65%, 50%, and 75%, respectively, using 40 mM acceptor and 200 mM sucrose as glucosyl donor. 3-Methoxylcatchol was also glycosylated, though at a significantly lower rate. A number of other aromatic compounds such as phenol, 2-hydroxybenzaldehyde, 1,3-dihydroxybenzene, and 1, 2 phenylethanediol were not glycosylated by GTF-D. Consequently GTF-D aromatic acceptors appear to require two adjacent aromatic hydroxyl groups. In order to facilitate the transglycosylation of less water-soluble acceptors the use of various water miscible organic solvents (cosolvents) was studied. The flavonoid catechin was used as a model acceptor. Bis-2-methoxyethyl ether (MEE) was selected as a useful cosolvent. In the presence of 15% (v/v) MEE the specific catechin transglucosylation activity was increased 4-fold due to a 12-fold increase in catechin solubility. MEE (10-30% v/v) could also be used to allow the transglycosylation of catechol, 4-methylcatechol, and 3-methoxycatechol at concentrations (200 mM) otherwise inhibiting GTF-D transglycosylation activity. PMID- 11005919 TI - Microbial sulfate reduction in a liquid-solid fluidized bed reactor. AB - A liquid-solid fluidized bed reactor was used to carry out sulfate reduction with a mixed culture of sulfate reducing bacteria. The bacteria were immobilized on porous glass beads. Stable fluidized bed operation with these biofilm-coated beads was possible. The low specific gravity of the hydrated beads allowed operation at low liquid recirculation rates. H(2)S level in the reactor was controlled by N(2) sparging, which also served as the location for liquid feed and removal. Ethanol was used as the electron donor/carbon source for the bacteria. Sulfate reduction rates up to 6.33 g sulfate L(-1) day(-1) were attained in the reactor at a hydraulic retention time of 5.1 h. The effect of hydraulic retention time and biomass loading on the beads, on reactor performance, and efficiency were examined. The efficiency of sulfate reduction increases considerably as the hydraulic retention increases, until the bacteria became very strongly substrate-limited at 55h HRT. The effect of bead biomass loading on bed expansion at various liquid superficial velocities was studied. A model for the reactor was developed. Simulations of the continuous flow experiments indicate that the model can describe the system well, and thus could be used in the design/scale-up of such reactors. The model suggests that a significant increase in the sulfate reduction capacity of the system is possible by increasing the volume of the bed relative to the total liquid volume of the system. PMID- 11005920 TI - Studies related to the scale-up of high-cell-density E. coli fed-batch fermentations using multiparameter flow cytometry: effect of a changing microenvironment with respect to glucose and dissolved oxygen concentration. AB - Multiparameter flow cytometric techniques developed in our laboratories have been used for the "at-line" study of fed-batch bacterial fermentations. These fermentations were done at two scales, production (20 m(3)) and bench (5 x 10(-3) m(3)). In addition, at the bench scale, experiments were undertaken where the difficulty of achieving good mixing (broth homogeneity), similar to that found at the production scale, was simulated by using a two-compartment model. Flow cytometric analysis of cells in broth samples, based on a dual-staining protocol, has revealed, for the first time, that a progressive change in cell physiological state generally occurs throughout the course of such fermentations. The technique has demonstrated that a changing microenvironment with respect to substrate concentration (glucose and dissolved oxygen tension [DOT]) has a profound effect on cell physiology and hence on viable biomass yield. The relatively poorly mixed conditions in the large-scale fermentor were found to lead to a low biomass yield, but, surprisingly, were associated with a high cell viability (with respect to cytoplasmic membrane permeability) throughout the fermentation. The small-scale fermentation that most clearly mimicked the large-scale heterogeneity (i.e., a region of high glucose concentration and low DOT analogous to a feed zone) gave similar results. On the other hand, the small-scale well-mixed fermentation gave the highest biomass yield, but again, surprisingly, the lowest cell viability. The scaled-down simulations with high DOT throughout and locally low or high glucose gave biomass and viabilities between. Reasons for these results are examined in terms of environmental stress associated with an ever increasing glucose limitation in the well-mixed case. On the other hand, at the large scale, and to differing degrees in scale-down simulations, cells periodically encounter regions of relatively higher glucose concentration. PMID- 11005921 TI - Degradation of phenol and benzoic acid in a three-phase fluidized-bed reactor. AB - Degradation of phenol and benzoic acid was studied in a fluidized-bed reactor (liquid volume 2.17 L) under nonsterile conditions with special emphasis on maximizing the flow through the reactor and investigating reactor performance at fluctuating feeds. Reactor response to substrate pulses was investigated by applying substrate square-wave inputs at a liquid flow of 1.00 L h(-1). A twofold increase of the phenol and benzoic acid feed concentrations for 2.5 h did not lead to accumulation and breakthrough. The cells were able to survive four to fivefold increases of the feed concentration for 1 h without loss of viability, although the phenol pulse lead to phenol accumulation in the reactor. Reactor performance at constantly fluctuating loads was investigated by varying the feed concentrations using sine wave functions. No accumulation of phenol or benzoic acid was observed. Influence of induction was studied using shift experiments. After 35 days of operation (369 hydrodynamic residence times) with phenol as sole substrate (carbon source) the reactor was able to mineralize benzoic acid without any adaptation or lag phase. The capability of phenol degradation, on the other hand, was lost by most cells after only 3 days operation with benzoic acid as the sole substrate. The experiments underline the importance of induction. In order to maximize the flow through the reactor, the liquid flow was increased stepwise while the feed concentrations were reduced correspondingly, keeping the volumetric conversion rates of phenol (0.24 g L(-1) h(-1)) and benzoic acid (0.17 g L(-1) h(-1)) constant. By this means, liquid flow could be increased up to 13.32 L h(-1), which was more than 20-fold higher than the maximum liquid flow achievable in a chemostat using the same conditions. PMID- 11005922 TI - Degradation of organophosphorous nerve agents by enzyme-polymer nanocomposites: efficient biocatalytic materials for personal protection and large-scale detoxification. AB - The biocatalytic destruction of organophosphates has become an important focus area, as efficient "clean" technologies are sought for chemical weapons decommissioning, counteracting nerve agent attacks, and protecting against organophosphate pesticide poisoning. A novel method is advanced for immobilizing the broad-spectrum enzyme organophosphorous hydrolase (OPH) from Pseudomonas diminuta, based on the formation of nanocomposite protein-silicone polymers. The resulting materials are highly active, stable, and versatile biocatalysts for the liquid and gas phase detoxification of organophosphates, and can be fabricated as monoliths, sheets, thick films, granulates, or macroporous foams. This approach offers an efficient avenue to robust, high-performance biocatalytic OPH containing polymers that outperform immobilized OPH catalysts reported to date. The method provides for the first time a route to biocatalytic materials that may be suitable for "active" protective wear, as well as bulk catalysts for the destruction of large volumes of organophosphates. The preparation of OPH-silicone biocomposites, their performances in the liquid and gas phase detoxification of paraoxon, dichlorvos, and diisopropyl fluorophosphate, and their features are discussed. PMID- 11005923 TI - Enzymatic synthesis of oligosaccharides: product removal during a kinetically controlled reaction. AB - In this article, the enzymatic production of oligosaccharides, which is an example of a kinetically controlled reaction, is studied. The aim is to show that the product yield can be enhanced by continuous removal of oligosaccharides from the reaction mixture. The oligosaccharides were removed by adsorption on activated carbon. The absorption could be described by the multicomponent Langmuir isotherm with different maximum saturation constants for mono-, di-, and trisaccharides. The affinity for trisaccharides was larger (k(tri) = 3.52 l/g) than for di- (k(di) = 0.94 l/g) and monosaccharides (k(mono) = 0.11 l/g). A model combining kinetics, adsorption on activated carbon, and mass transfer in an adsorption column was developed. Model calculations for the batch process with removal showed a yield improvement of 23% compared to the batch process without removal. Experimentally, a yield improvement of 30% was obtained. Model calculations for the continuous process studied did not result in an increase of the yield. The advantages of removal were masked by the negative influence of recirculation and the relative large time between formation and removal. PMID- 11005924 TI - Effect of insulin stimulation on the proliferation and death of Chinese hamster ovary cells. AB - The effect of environmental and genetic parameters on cell death was studied in Chinese hamster ovary cell cultures. Experiments were performed using an anchorage-dependent CHO cell line expressing gamma-IFN, and a second cell line obtained by transfection of the previous one with bcl-2. In serum-free medium the two cell lines showed a considerable degree of growth control entering quiescence while maintaining high viabilities. The addition of transferrin did not have any effect but insulin addition allowed cells arrested by serum withdrawal to reenter the cell cycle. However, insulin supplementation also resulted in cell death, which was possible to avoid through bcl-2 overexpression or in the presence of serum. We propose that the expression of c-myc, shown to be induced by insulin, plays an important role in the cell death observed after insulin addition in an inappropriate environment, deficient in protective factors. This hypothesis is supported by measurements of c-myc expression and cell cycle distribution. PMID- 11005925 TI - Modeling substrate interactions during the biodegradation of mixtures of toluene and phenol by Burkholderia species JS150. AB - The biodegradation kinetics of toluene, phenol, and a mixture of toluene and phenol by Burkholderia species JS150 was measured and modeled. Both of these compounds can serve as the sole source of carbon and energy for this microorganism. The single-substrate biodegradation kinetics was described well using the Monod model, with model constants of mu(max,T) = 0.39 h(-1) and K(S,T) = 0.011 mM for growth on toluene and mu(max,P) = 0.309 h(-1) and K(S,P) = 0.0054 mM for growth on phenol. Degradation of the mixture of toluene and phenol followed simultaneous utilization kinetics with toluene being the preferred substrate. Toluene was found to inhibit the rate of utilization of phenol while the presence of phenol had little effect on the rate of degradation of toluene. Of the kinetic models that were tested, one developed for microbial degradation of multiple substrates was able to describe substrate interactions and to model the mixture utilization by strain JS150. Simple competitive, noncompetitive, or uncompetitive substrate kinetics were not sufficient to describe the observed inhibitory interactions. PMID- 11005926 TI - Use of 16S-rRNA to investigate microbial population dynamics during biodegradation of toluene and phenol by a binary culture. AB - Interspecies interactions and changes in the rate and extent of biodegradation in mixed culture-mixed substrate studies were investigated. A binary mixed culture of Pseudomonas putida F1 and Burkholderia sp. JS150 degraded toluene, phenol, and their mixture. Both toluene and phenol can serve as sole sources of carbon and energy for both P. putida F1 and strain JS150. To investigate the population dynamics of this system, a fluorescent in-situ hybridization method was chosen because of its ability to produce quantitative data, its low standard error, and the ease of use of this method. When the binary mixed culture was grown on toluene or phenol alone, significant interactions between the species were observed. These interactions could not be explained by a pure-and-simple competition model and were substrate dependent. Strain JS150 growth was slightly inhibited when grown with P. putida F1 on phenol, and P. putida F1 grew more rapidly than expected. Conversely, when the two species were grown together on toluene alone, P. putida F1 was inhibited while strain JS150 was unaffected. During growth of the mixed culture on a combination of toluene and phenol, the interactions were similar to that observed during growth on phenol alone; P. putida F1 growth was enhanced while strain JS150 was unaffected. Because of the observed interspecies interactions, monoculture kinetic parameters were not sufficient to describe the mixed culture kinetics in any experiment. This is one of the first reports of microbial population dynamics in which molecular microbial ecology and mathematical modeling have been combined. The use of the 16S-rRNA-based method allowed for observation and understanding of interspecies interactions that were not observable with standard culture-based methods. These results suggest the need for more investigations that account for both substrate and microbial interactions when predicting the fate of organic pollutants in real systems. PMID- 11005927 TI - Error-prone PCR of Vitreoscilla hemoglobin (VHb) to support the growth of microaerobic Escherichia coli. AB - Expression of the gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been previously used to improve recombinant cell growth and enhance product formation under microaerobic conditions. It is very likely that the properties of VHb are not optimized for foreign hosts; therefore, we used error-prone PCR to generate a number of randomly mutated vhb genes to be expressed and studied in Escherichia coli. In addition, the mutated VHb proteins also contained an extension of eight residues (MTMITPSF) at the amino terminus. VHb mutants were screened for improved growth properties under microaerobic conditions and 15 clones expressing mutated hemoglobin protein were selected for further characterization and cultivated in a microaerobic bioreactor to analyze the physiological effects of novel VHb proteins on cell growth. The expression of four VHb mutants, carried by pVM20, pVM50, pVM104, and pVM134, were able to enhance microaerobic growth of E. coli by approximately 22%, 155%, 50%, and 90%, respectively, with a concomitant decrease of acetate excretion into the culture medium. The vhb gene in pVM20 contains two mutations substituting residues Glu19(A17) and Glu137(H23) to Gly. pVM50 expresses a VHb protein carrying two mutations: His36(C1) to Arg36 and Gln66(E20) to Arg66. pVM104 and pVM134 express VHb proteins carrying the mutations Ala56(E10) to Gly and Ile24(B5) to Thr, respectively. Our experiments also indicate that the positive effects elicited by mutant VHb-expression from pVM20 and pVM50 are linked to the peptide tail. Removal of the N-terminal sequence reduced cell growth approximately 23% and 53%, respectively, relative to wild-type controls. These results clearly demonstrate that it is possible to obtain mutated VHb proteins with improved characteristics for improving microaerobic growth of E. coli by using combined mutation techniques, addition of a peptide tail, and random error-prone PCR. PMID- 11005928 TI - Construction and structural modeling of a single-chain Fv-asparaginase fusion protein resistant to proteolysis. AB - In this study, we construct a fusion protein composed of L-asparaginase (ASNase; from Escherichia coli AS 1.357) and a protective single-chain Fv (scFv), which was selected from a phage-display scFv library from our previous studies. The antibody moiety of the fusion protein was fused to the N-terminus of the enzyme moiety via a linker peptide, (Gly(4)Ser)(6). Recombinant plasmid pET-SLA was constructed to express scFv-ASNase fusion to high levels in E. coli and the expressed product was found to form inclusion bodies. We obtained a soluble fusion protein by refolding and purification. The soluble fusion protein exhibited about 82% of the enzymatic activity of the native ASNase at the same molar concentration, and had a K(m) value similar to that of the native enzyme for the substrate L-asparagine. Importantly, the fusion protein was more stable than native ASNase. In addition: (1) following treatment with trypsin, alpha chymotrypsin, and rennet, at 37 degrees C for 30 min, scFv-ASNase fusion retained 94.0%, 88.8%, and 84.5% of its original activity, respectively, whereas native ASNase became inactive; and (2) ScFv-ASNase fusion had a much longer in vitro half-life (9 h) in serum than the native enzyme (2 h). The three-dimensional structure of the fusion protein was obtained by modeling with the Homology and Discover modules of the INSIGHT II software package. On the basis of the structural evidence and biochemical properties, we propose that the scFv moiety of the fusion protein may confer ASNase moiety resistance to proteolysis as a result of both steric hindrance and a change in the electrostatic surface of the enzyme. PMID- 11005929 TI - High density cultivation of two strains of iron-oxidizing bacteria through reduction of ferric iron by intermittent electrolysis. AB - Electrolytic cultivation was applied to Leptospirillum ferrooxidans strains P3A and CF27, which use ferrous iron to respire aerobically. Ferrous iron was supplied to the bacteria by intermittent electrolytic reduction of ferric iron as electron shuttle using an electrode. The yield of L. ferrooxidans and strain CF27 reached 20- and 50-fold, respectively, higher density than were achievable yields without electrolysis. The time required to obtain high density depended not on the growth ratio, but rather on the original growth rate of each strain. PMID- 11005930 TI - The inhibition of Escherichia coli lac operon gene expression by antigene oligonucleotides-mathematical modeling. AB - Gene transcription is regulated by transcription factors that can bind to specific regions on DNA. Antigene oligonucleotides (oligos) can bind to specific regions on DNA and form a triplex with the double-stranded DNA. The triplex can competitively inhibit the binding of transcription factors and, as a result, transcription can be inhibited. A genetically structured model has been developed to quantitatively describe the inhibition of the Escherichia coli lac operon gene expression by triplex-forming oligos. The model predicts that the effect of triplex-forming oligos on the lac operon gene expression depends on their target sites. Oligonucleotides targeted to the operator are much more effective than those targeted to other regulatory sites on the lac operon. In some cases, the effect of oligo binding is similar to that of a mutation in the lac operon. The model provides insight as to the specific binding site to be targeted to achieve the most effective inhibition of gene expression. The model is also capable of predicting the oligo concentration needed to inhibit gene expression, which is in general agreement with results reported by other investigators. PMID- 11005931 TI - All solid-state GFP sensor. AB - An all-solid-state green fluorescent protein (GFP) sensor for GFP measurement was developed. It is immune to interference from ambient light and works with standard flow-through cuvettes. The sensor is practically insensitive to the scattered excitation light encountered in microbial suspensions. It has a range of 0.0002-1 g/L (7.4 x 10(-9) - 3.7 x 10(-5) M) with limit of detection 0.00019 g/L (7.0 x 10(-9) M). The sensor could be used with a UV or blue light emitting diode (LED) as a light source, depending on required sensitivity, selectivity, and background levels. Its very low cost makes it useful in a variety of applications. This article describes the construction and validation of the sensor both off- and on-line in fermentation processes. PMID- 11005932 TI - Oral glycoprotein IIb/IIIa antagonists: new insights from the SYMPHONY trial. AB - Platelet aggregation plays a central role in the pathophysiology of acute coronary syndromes, and the platelet glycoprotein IIb/IIIa receptor has been identified as the critical final mediator of this process. Antagonists of this receptor used parenterally during both acute coronary syndromes and percutaneous coronary interventions reduce the likelihood of subsequent major cardiac complications. However, after the treatment period little further benefit accrues. Based on these observations and that of the significant benefit of aspirin in cardiovascular secondary prevention, oral glycoprotein IIb/IIIa receptor antagonists are being evaluated with the goal of extending the benefit of glycoprotein IIb/IIIa inhibition into chronic secondary prevention. This paper will review the results of the SYMPHONY study of one such oral agent, sibrafiban, and the current state of the oral glycoprotein IIb/IIIa inhibitor field. PMID- 11005933 TI - Clopidogrel and coronary stenting: what is the next question? AB - Today, following coronary stenting, clopidogrel has largely replaced ticlopidine as part of combination antiplatelet therapy following coronary stenting primarily due to its better tolerability. While there is no randomized, blinded, efficacy trial of ticlopidine versus clopidogrel, there are ample data from a number of observational studies, randomized non-blinded trials, and a randomized blinded safety trial to prove that clopidogrel is not only safer than ticlopidine, but also at least as efficacious following stenting. With over 10,000 treated patients, pooled data suggest similar rates of stent thrombosis (clopidogrel 0.98% vs. ticlopidine 0.98%) and lower rates of major adverse cardiac events with clopidogrel (clopidogrel 1.63% vs. ticlopidine 4.52%, p<0.001), with a clear advantage for clopidogrel regarding adverse events (clopidogrel 5.91% vs. ticlopidine 9.75%, p<0.001). With clopidogrel's superior safety and at least equivalent efficacy, the question of "which thienopyridine?" post-stenting has been answered. Now the questions "how much?", "how soon?" and "how long?" must be addressed. The Clopidogrel for the Reduction of Events During Observation (CREDO) trial is a multi-center, double-blind, randomized trial designed to answer these remaining questions. CREDO will evaluate the efficacy and safety of clopidogrel pretreatment versus no pretreatment, and prolonged (1 year), versus short-term (1 month) dual antiplatelet therapy in 2,000 patients undergoing planned or highly probable coronary intervention with a stent. PMID- 11005934 TI - A pharmacodynamic study of clopidogrel in chronic hemodialysis patients. AB - Combination antiplatelet agents, particularly aspirin and ticlopidine, have found increased use in the prevention of arterial thrombosis. Clopidogrel, a thienopyridine derivative, like ticlopidine was recently approved by the U.S. Food and Drug Administration (FDA) for the reduction of ischemic events in patients with myocardial infarction, stroke, or peripheral arterial disease and appears to have much less hematologic toxicity than ticlopidine has. Thrombosis of hemodialysis access grafts is a major cause of morbidity in this patient population. Combination antiplatelet agents may be particularly useful in the prevention of hemodialysis access graft thrombosis. In preparation for such a study, we have performed a pharmacodynamic study of the platelet inhibitory effects of clopidogrel in patients on maintenance hemodialysis. Nine chronic hemodialysis patients were studied. Baseline platelet aggregation studies were performed, after which the subjects were begun on clopidogrel 75 mg daily. Platelet aggregation studies were repeated after 14 days of therapy. Drug was stopped and a final set of platelet aggregation studies were performed 7 days later. Because clopidogrel acts by inhibiting adenosine diphosphate (ADP)-induced platelet aggregation, we used ADP as the agonist in the platelet aggregation studies. We also measured the time required to achieve hemostasis after removing the dialysis needles at the termination of a dialysis session. Patients were carefully monitored for any adverse reaction to clopidogrel. Fourteen days' treatment with clopidogrel inhibited ADP-induced platelet aggregation from 48 to 23% with ADP 2 microM (P=0.0113), from 59 to 38% with ADP 5 microM (P=0. 0166), and from 66 to 44% with ADP 10 microM (P=0. 0172). This inhibition of platelet aggregation was reversed 7 days after stopping clopidogrel. Clopidogrel administration did not affect the time required to achieve hemostasis after removal of the dialysis needles. No adverse reactions were noted. No patient had evidence of bleeding, rash or gastro-intestinal (GI) upset. Clopidogrel inhibits ADP-induced platelet aggregation in subjects receiving chronic maintenance hemodialysis. The magnitude of inhibition is similar to that reported in nonuremic subjects with atherosclerosis. This inhibition is reversible within 7 days of discontinuing the drug. No adverse reactions to the drug were noted in this short-term (14-day) trial. PMID- 11005935 TI - Prevalence of early left ventricular thrombus after primary coronary intervention for acute myocardial infarction. AB - The prevalence of left ventricular (LV) thrombus after acute myocardial infarction (AMI) has been reported high at 20-60%. Current reperfusion therapies such as thrombolysis have shown a trend toward reducing the incidence of LV thrombosis. However, the prevalence of LV thrombus after primary percutaneous coronary intervention (PCI) for AMI has not been systematically studied. At Massachusetts General Hospital 71 consecutive patients who underwent primary PCI for acute ST elevation MI were reviewed for the prevalence of LV thrombus evaluated by echocardiography. Echocardiography was performed within 5 days of infarction. PCI was successful in all patients. The time delay from symptom onset to intervention was 191 minutes. Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow was achieved in more than 80% of cases. Only 3 patients (4%) had echocardiographic evidence of LV thrombus. All 3 patients had anterior infarctions. The incidence among patients with anterior MI was 10% (3 of 30 patients). The prevalence of LV thrombus in patients treated with primary PCI for AMI is low (4%). PMID- 11005936 TI - The use of the electrocardiogram to identify epicardial coronary and tissue reperfusion in acute myocardial infarction. AB - The standard 12-lead electrocardiogram (ECG) gives us crucial information concerning myocardial perfusion and the success of reperfusion therapy for ST elevation acute myocardial infarction. Continuous monitoring has advantages over repeated snapshot recordings. There are four electrocardiographic markers for prediction of the perfusion status of the ischemic myocardium: (1) ST-segment measurements, (2) T-wave configuration, (3) QRS changes, and (4) reperfusion arrhythmias. Complete and stable (> or = 70%) resolution of ST-segment elevation is associated with better outcome and preservation of left ventricular function than partial (30 to 70%) or no (<30%) ST-segment resolution. Early inversion of the T waves after initiation of reperfusion therapy is another marker of myocardial reperfusion and a good prognostic sign. Using standard 12-lead ECG, dynamic changes in Q-wave number, amplitude, and width; R-wave amplitude; and S wave appearance are detected during reperfusion therapy. However, the significance of these changes has not been clarified. Reperfusion arrhythmias, especially bradycardia and accelerated idioventricular rhythm, are detected occasionally during reperfusion therapy, but the value of reperfusion arrhythmias as a marker of coronary artery patency is still debatable. Dynamic changes in the QRS complexes, ST segments and T waves occur during reperfusion therapy and the days after. Whereas changes in ST-segment amplitude have been extensively studied, the significance of QRS-complex and T-wave changes is less clear, and especially whether changes in the QRS complex and T wave may be complementary and additive to ST-segment monitoring. It has remained unclear whether electrocardiographic signs of reperfusion and reischemia should be used for therapeutic decision making in the clinical setting. PMID- 11005937 TI - Temporary reversal of anticoagulation using oral vitamin K. AB - Brief reversal of oral anticoagulant therapy is frequently necessary prior to minor surgery or invasive procedures. We sought to determine the effect of an oral dose of 2.0 mg of vitamin K(1) on the international normalized ratio (INR) among patients with a stable therapeutic INR who were maintained on their daily dose of warfarin. We prospectively studied a convenience cohort of patients attending an anticoagulation clinic who had either just completed treatment for venous thromboembolism or were receiving prophylaxis for atrial fibrillation, cardiomyopathy, or peripheral vascular disease. Each patient received an oral dose of 2.0 mg of aqueous vitamin K(1). Serial INR measurements were taken over 1 week. There was wide variation in the INR response between patients, from no change to complete reversal of anticoagulation. The effect also varied widely over time. There was a significant inverse correlation between the fall in logarithm of the INR and the daily warfarin dose required to achieve an INR value of 2.5 (r=-0.52, p=0.011). Use of a 2.0 mg oral dose of vitamin K(1) does not reliably reverse (correct) a therapeutic INR in patients maintained on their daily dose of warfarin. PMID- 11005939 TI - Epidemiology, diagnosis, and management of cardiac disease in chronic renal disease. AB - There is an extremely high burden of cardiovascular disease (CVD) in patients with renal disease. Both traditional as well as uremia-related factors are contributory. Diagnosis of CVD has limitations in patients with renal disease, and suspicion for the presence of CVD needs to be high even in the absence of classic symptoms. Prevention and management of CVD is similar to the general population but important differences need to be noted. PMID- 11005938 TI - Drug-induced thrombocytopenia in the coronary care unit. AB - Drug-induced thrombocytopenia is a phenomenon that causes significant morbidity and mortality among patients. Practitioners should be able to recognize the clinical manifestations of drug-induced thrombocytopenia, differentiate it from other causes, and manage it appropriately. Numerous case reports have documented drug-induced causes of thrombocytopenia. The following article focuses on the characteristics and management of drug-induced thrombocytopenia secondary to medications commonly encountered in the coronary care unit. Pharmacotherapeutic agents that are most commonly implicated in this setting include ticlopidine, unfractioned heparin, glycoprotein (GP)IIb/IIIa inhibitors, H(2)-receptor antagonists, quinidine and antibiotics. Case reports were obtained through a comprehensive search of the Medicine database and subsequently complemented by bibliographic reviews of the agents just specified. Reports that exhibited possible, probable, and definite associations with drug-induced thrombocytopenia are included in the article. PMID- 11005940 TI - What is the required reperfusion period for assessment of myocardial infarct size using triphenyltetrazolium chloride staining in the rat? AB - Measurements of infarct size by use of tripenyltetrazoliumchloride (TTC) is a widely accepted method used to delineate the extent of myocardial necrosis following coronary occlusion and reperfusion in various animal experiments. There is controversy, however, regarding the optimal reperfusion time for estimating the maximally infarcted area by TTC staining in the rat. We tested six different reperfusion times following 90 minutes of regional myocardial ischemia. Group 1 had 5 minutes of reperfusion (n=6), group 2 had 30 minutes of reperfusion (n=6), group 3 had 1 hour of reperfusion (n=6), group 4 had 2 hours of reperfusion (n=6), group 5 had 3 hours of reperfusion (n=6), group 6 had 4.5 hours of reperfusion (n=6). Risk areas, measured by the use of blue dye, were similar among the 6 study groups. Infarct size as a percent of risk area was 57+/-11% in group 1, 74+/-7% in group 2, 61+/-9% in group 3, 71+/-5% in group 4, 70+/-5% in group 5, and 64+/-9% in group 6 (x+/-standard error [SE]). There was no significant difference in infarct size between the groups. However, prior to 60 minutes of reperfusion, patches of pink and white areas were observed within the risk regions, indicating a more difficult assessment of proper delineation of outer regions of necrotic tissue. For acute assessment of infarct size, reperfusion for 60 minutes or more is optimal because the infarct does appear homogeneous at that time and does not become larger with longer reperfusion periods. PMID- 11005942 TI - Effect of coronary thrombolysis on the plasma concentration of osteonectin (SPARC, BM40) in patients with acute myocardial infarction. AB - Osteonectin is a phosphoglycoprotein exclusively located in bone and platelet alpha-granules. Human platelet-derived osteonectin is released into plasma after thrombin-induced activation. Recognizing the unique distribution of the osteonectin pool, we first sought to investigate whether osteonectin could serve as a sensitive marker of platelet activity, and identify patients with acute myocardial infarction (AMI). The second objective was to define the effects of thrombolytic therapy in these patients on the plasma concentrations of osteonectin at prespecified time points following attempted reperfusion. Osteonectin levels by ELISA were determined in AMI patients before thrombolysis and at 3, 6, 12, and 24 hours thereafter and compared with 12 healthy controls. At baseline, soluble osteonectin plasma levels were similar between controls (447. 7+/-20.6 ng/ml) and AMI patients (425.7+/-43.3 ng/mL; p=NS). A significant increase of the soluble osteonectin was observed at 3 hours after thrombolysis (519.4+/-26.9 ng/mL; p=0.03), and was followed by a decrease to baseline levels at 6 hours after attempted reperfusion. Contrary to expectations, the plasma osteonectin level in our pilot study was not a sensitive marker distinguishing patients with AMI. The early peak of soluble osteonectin at 3 hours after thrombolytic therapy is most likely not related to coronary thrombolysis per se but rather to the phasic changes of platelet activity during myocardial ischemia reperfusion. The unquestionable platelet origin of this protein and the lack of elevated plasma levels of this alpha-granule constituent, challenge the postulate of uniform platelet activation in AMI patients. PMID- 11005941 TI - Combination of a direct thrombin inhibitor and a platelet glycoprotein IIb/IIIa blocking peptide facilitates and maintains reperfusion of platelet-rich thrombus with alteplase. AB - We sought to determine the efficacy of the combination of argatroban, a direct thrombin inhibitor, and G4120, a platelet glycoprotein (GP) IIb/IIIa blocker, to enhance thrombolysis with alteplase. Platelet-rich thrombus in the rabbit arterial thrombosis model is relatively resistant to alteplase despite the addition of aspirin and heparin. The adjunctive use of either direct thrombin inhibitors or GP IIb/IIIa inhibitors in thrombolysis has been investigated with encouraging, but limited, success. The usefulness of combining both agents as adjunctive therapy to thrombolysis has not been fully explored. Following platelet-rich thrombus formation in the rabbit, argatroban (3 mg/kg), G4120 (0.5 mg/kg), G4120 plus heparin (200 U/kg), or G4120 plus argatroban were intravenously infused over 60 minutes. Alteplase was given as intravenous boluses (0.45 mg/kg) at 15-minute intervals up to 4 doses or until reperfusion. Blood flow and bleeding time were monitored for 2 hours. The combination of G4120 plus argatroban resulted in a persistent patency in 5 of 7 animals compared with 0 of 6 for argatroban alone (p=0.02), 1 of 6 for G4120 alone (p=0.08), and 2 of 6 for G4120 plus heparin (p=0.2). Although during the infusion the bleeding times were longer in the groups that received G4120 (26+/-7.7 minutes vs. 14+/-10 minutes, p<0.05), by the end of the experiment there were no statistically significant differences. Similarly, during the infusion the activated partial thromboplastin times (aPTT) was higher in groups that received heparin or argatroban (99+/-51 seconds vs. 32+/-7.6 seconds, p<0.001), but by the end of the experiment the aPTTs had returned to close to baseline in all groups except the G4120 plus heparin group. These results suggest that lysis of platelet-rich thrombus with alteplase requires the addition of both potent platelet and thrombin inhibitors. Specifically designed agents, G4120 and argatroban, are effective without additional increased risk for bleeding. PMID- 11005943 TI - Changes in movement variables associated with transient overshoot of the final position. AB - Transient overshoot (TO), which is assessed as the distance between the movement amplitude and the final position, was measured in a series of rapid, discrete elbow flexion movements performed under different distance and loading conditions by 7 participants. A positive relationship was found between kinematic variables (peak velocity, peak acceleration and deceleration, and the symmetry ratio) and the magnitude of TO, particularly in short movements performed against a light load. The relationships between TO and electromyographic (EMG) variables were low and mainly insignificant. Thus, TO contributes to the variability of rapid, discrete movements and therefore should be taken into account as an additional parameter in studies of the scaling of movement variables with movement mechanical conditions. TO could also represent a consequence of mechanical properties of the single-joint system rather than an independently programmed primary submovement. PMID- 11005944 TI - Developmental features of rapid aiming arm movements across the lifespan. AB - Using a lifespan approach, the authors investigated developmental features of the control of ballistic aiming arm movements by manipulating movement complexity, response uncertainty, and the use of precues. Four different age groups of participants (6- and 9-year-old boys and girls and 24- and 73-year-old men and women, 20 participants in each age group) performed 7 types of rapid aiming arm movements on the surface of a digitizer. Their movement characteristics such as movement velocity, normalized jerk, relative timing, movement linearity, and intersegment intervals were profiled. Analyses of variance with repeated measures were conducted on age and task effects in varying movement complexity (Study 1), response uncertainty (Study 2), and precue use (Study 3) conditions. Young children and senior adults had slower, more variant, less smooth, and less linear arm movements than older children and young adults. Increasing the number of movement segments resulted in slower and more variant responses. Movement accuracy demands or response uncertainty interacted with age so that the 6- and 74-year-old participants had poorer performances but responded similarly to the varying treatments. Even though older children and young adults had better performances than young children and senior adults, their arm movement performance declined when response uncertainty increased. The analyses suggested that young children's and senior adults' performances are poorer because less of their movement is under central control, and they therefore use on-line adjustments. In addition, older children and young adults use a valid precue more effectively to prepare for subsequent movements than do young children and senior adults, suggesting that older children and young adults are more capable of organizing motor responses than are young children and senior adults. PMID- 11005945 TI - Variability and noise in continuous force production. AB - In the present 3 experiments, the authors examined the hypothesis, derived from information theory, that increases in the variability of motor responses result from increases in perceptual-motor noise. Three different groups of participants (Ns = 10, 9, and 10, respectively, in Experiments 1, 2, and 3) produced continuous isometric force under either low, intermediate, or high target force levels. When considered together, the results showed that force variability (SD) increased exponentially as a function of force level. However, an index of information transmission (M/SD), as well as measures of noise in both the time (approximate entropy) and the frequency (power spectrum) domains, changed according to an inverted-U-shaped function over the range of force levels. The findings provide further evidence that increased information transmission is related to increases, and not to decreases, in the noisiness of the structure of force output. PMID- 11005946 TI - Performance and learning of generalized motor programs: relative (GMP) and absolute (parameter) errors. AB - The effects of practice (Experiment 1) and parameter variability (Experiment 2) on the learning of generalized motor programs (GMPs) and movement parameterization were investigated. In each experiment, 2 tasks with different relative force-time structures were tested. Participants (N = 32, Experiment 1; N = 40, Experiment 2) attempted to exert a pattern of force that resembled in force and time a waveform that was displayed on a computer monitor. In both experiments, the analysis suggested that the GMP, although refined over practice, was relatively stable (i.e., resistant to decay and interference), even early in practice (after 20 trials). In addition, the results indicated that constant and variable parameter practice did not differentially affect GMP learning but did degrade the learning of the parameter that was not varied. The data provided additional evidence for the dissociation of the GMP and the parameterization processes proposed in GMP theory. Contrary to schema theory, the present data suggest an interdependence between the force and the time parameters: The manipulation of 1 of the parameters has a negative effect on the learning of the other parameter. PMID- 11005947 TI - Bimanual finger tapping: effects of frequency and auditory information on timing consistency and coordination. AB - The authors' goal in this study was to probe the basis for an earlier, unexpected finding that preferred-frequency finger tapping tends to have higher frequencies and to be less stable for in-phase than for antiphase tasks. In follow-up experiments, 3 protocols were employed: a preferred-frequency replication in both coordination modes, a metronome-driven matching of the preferred frequencies to each of the coordination modes, and a frequency scaling of both modes. The original findings were affirmed for preferred frequency. Tapping to a metronome had a differential effect on in-phase and antiphase: A more stable coupling across frequencies was exhibited during in-phase. Under frequency scaling, the antiphase pattern decomposed at lower frequencies than did in-phase, but no phase transitions were observed. The loss of stable coordination in both modes was attended by sudden increases in frequency differences between fingers and by phase wandering. The emergence of those effects is discussed in light of asymmetric modifications to the Haken-Kelso-Bunz model (H. Haken, J. A. S. Kelso, & H. Bunz, 1985) and the task constraints of tapping. PMID- 11005948 TI - Temporal precision in tapping and circle drawing movements at preferred rates is not correlated: further evidence against timing as a general-purpose ability. AB - Recently, researchers have discovered that individuals who are consistent timers in a tapping task are not necessarily consistent timers when they perform a continuous drawing task. In other words, nonsignificant correlations were found among tapping and drawing movements for timing precision (S. D. Robertson et al., 1999). In the present experiment, the authors investigated whether or not consistency in timing for tapping and drawing was correlated when participants (N = 24) were allowed to move at their preferred rate of movement. There were no significant correlations between tapping and drawing in terms of timing precision. That result lends further support to the notion that timing behavior is specific to the nature of the task, and thus further weakens the idea that timing is a generalized ability that can be imposed on a variety of different types of tasks. PMID- 11005949 TI - Adaptation of neuromuscular synergies during intentional constraints of space time relationships in human gait. AB - Tight frequency-to-amplitude relationships are observed in spontaneous human steady gait. They can be modified, if required; that flexibility forms a fundamental basis of the intentional adaptive capabilities of locomotion. In the present experiments, the processes underlying that flexibility were investigated at both the level of joint kinematics and the level of neuromuscular synergies. Subjects (N = 4) walked at the same speed either with a preferred or a nonpreferred frequency-to-amplitude relationship (i.e., constrained, short steps at a high frequency [COS condition] or constrained, long steps at a low frequency [COL condition]); their swing and stance phases were separately analyzed. In the COS condition, increases in EMG activity were specifically required during the swing phase. In the COL condition, several muscles required increases in EMG activity during the stance phase, but decreases of the hamstring muscles were needed during the swing phase. Whereas, in preferred walking, modification of the frequency affects the EMG patterns globally (the gain increasing with the frequency in both the stance and swing phases), the present results show that changing the frequency in a constrained manner either affects the swing phase specifically or affects both phases, but in the opposite direction. That finding indicates that a separate control is needed in both the swing and the stance phases. PMID- 11005950 TI - Geometric phases for mixed states in interferometry. AB - We provide a physical prescription based on interferometry for introducing the total phase of a mixed state undergoing unitary evolution, which has been an elusive concept in the past. We define the parallel transport condition that provides a connection form for obtaining the geometric phase for mixed states. The expression for the geometric phase for mixed state reduces to well known formulas in the pure state case when a system undergoes noncyclic and unitary quantum evolution. PMID- 11005951 TI - Exact particle and kinetic-energy densities for one-dimensional confined gases of noninteracting fermions. AB - We propose a new method for the evaluation of the particle density and kinetic pressure profiles in inhomogeneous one-dimensional systems of noninteracting fermions, and apply it to harmonically confined systems of up to N = 1000 fermions. The method invokes a Green's function operator in coordinate space, which is handled by techniques originally developed for the calculation of the density of single-particle states from Green's functions in the energy domain. In contrast to the Thomas-Fermi approximation, the exact profiles show negative local pressure in the tails and a prominent shell structure which may become accessible to observation in magnetically trapped gases of fermionic alkali atoms. PMID- 11005952 TI - Analytical results for the interaction energy of a trapped, weakly interacting Bose-Einstein condensate. AB - We examine the problem of a repulsive, weakly interacting and harmonically trapped Bose-Einstein condensate under rotation. We derive a simple analytic expression for the energy incorporating the interactions when the angular momentum per particle is between zero and one and find that the interaction energy decreases linearly as a function of the angular momentum in agreement with previous numerical and limiting analytical studies. PMID- 11005953 TI - Vortex precession in Bose-Einstein condensates: observations with filled and empty cores. AB - We have observed and characterized the dynamics of singly quantized vortices in dilute-gas Bose-Einstein condensates. Our condensates are produced in a superposition of two internal states of 87Rb, with one state supporting a vortex and the other filling the vortex core. Subsequently, the state filling the core can be partially or completely removed, reducing the radius of the core by as much as a factor of 13, all the way down to its bare value of the healing length. The corresponding superfluid rotation rates, evaluated at the core radius, vary by a factor of 150, but the precession frequency of the vortex core about the condensate axis changes by only a factor of 2. PMID- 11005954 TI - Supercooling molecular hydrogen down through the superfluid transition. AB - Recent calculations by Vorobev and Malyshenko [JETP Lett. 71, 39 (2000)] show that molecular hydrogen may stay liquid and superfluid in strong electric fields of the order of 4x10(7) V/cm. I demonstrate that strong local electric fields of similar magnitude exist beneath a two-dimensional layer of electrons localized in the image potential above the surface of solid hydrogen. Even stronger local fields exist around charged particles (ions or electrons) if the surface or bulk of a solid hydrogen crystal is statically charged. Measurements of the frequency shift of the 1 --> 2 photoresonance transition in the spectrum of a two dimensional layer of electrons above a positively or negatively charged solid hydrogen surface performed in the temperature range 7-13.8 K support the prediction of electric field induced surface melting. The range of surface charge density necessary to stabilize the liquid phase of molecular hydrogen at the temperature of superfluid transition is estimated. PMID- 11005955 TI - Fracture and second-order phase transitions. AB - Using the global fiber bundle model as a tractable scheme of progressive fracture in heterogeneous materials, we define the branching ratio in avalanches as a suitable order parameter to clarify the order of the phase transition occurring at the collapse of the system. The model is analyzed using a probabilistic approach suited to smooth fluctuations. The branching ratio shows a behavior analogous to the magnetization in known magnetic systems with second-order phase transitions. We obtain a universal critical exponent beta approximately = 0.5 independent of the probability distribution used to assign the strengths of individual fibers. PMID- 11005956 TI - Measurement of Newton's constant using a torsion balance with angular acceleration feedback. AB - We measured Newton's gravitational constant G using a new torsion balance method. Our technique greatly reduces several sources of uncertainty compared to previous measurements: (1) It is insensitive to anelastic torsion fiber properties; (2) a flat plate pendulum minimizes the sensitivity due to the pendulum density distribution; (3) continuous attractor rotation reduces background noise. We obtain G = (6.674215+/-0.000092) x 10(-11) m3 kg(-1) s(-2); the Earth's mass is, therefore, M = (5.972245+/-0.000082) x 10(24) kg and the Sun's mass is M = (1.988435+/-0.000027) x 10(30) kg. PMID- 11005957 TI - Fields over unsharp coordinates. AB - It has been shown that space-time coordinates can exhibit only very few types of short-distance structures, if described by linear operators: they can be continuous, discrete, or "unsharp" in one of two ways. In the literature, various quantum gravity models of space-time at short distances point towards one of these two types of unsharpness. Here, we investigate the properties of fields over such unsharp coordinates. We find that these fields are continuous--but possess only a finite density of degrees of freedom, similar to fields on lattices. As a special case we recover the Shannon sampling theorem of information theory. PMID- 11005958 TI - Search for lepton flavor violation in K+ decays into a charged pion and two leptons. AB - A search for lepton flavor violating decays, K+ --> mu+ mu+ pi-, K+ --> e+ e+ pi , K+ --> pi+ e+ mu-, K+ --> mu+ e+ pi-, and pi0 --> e+ mu-, was performed using the data collected in Experiment E865 at the Brookhaven Alternating Gradient Synchrotron. No signal was found in any of the decay modes. At the 90% confidence level, the branching ratios are less than 3.0x10(-9), 6.4x10(-10), 5. 2x10(-10), 5.0x10(-10), and 3.4x10(-9), respectively. PMID- 11005961 TI - Dynamical symmetry breaking in the sea of the nucleon. AB - We derive the nonanalytic chiral behavior of the flavor asymmetry d - u. Such behavior is a unique characteristic of Goldstone boson loops in chiral theories, including QCD, and establishes the unambiguous role played by the Goldstone boson cloud in the sea of the proton. Generalizing the results to the SU(3) sector, we show that strange chiral loops require that the s - s distribution be nonzero. PMID- 11005959 TI - Study of charmless hadronic B meson decays to pseudoscalar-vector final states. AB - We report results of searches for charmless hadronic B meson decays to pseudoscalar( pi(+/-), K+/-, pi(0), or K(0)(S))-vector( rho, K(*), or omega) final states. By using 9.7x10(6) BB pairs collected with the CLEO detector, we report the first observation of B(-)--->pi(-)rho(0), B(0)-->pi(+/-)rho(-/+), and B(-)-->pi(-)omega, which are expected to be dominated by hadronic b-->u transitions. The measured branching fractions are (10.4(+3.3)(-3.4)+/-2.1)x10( 6), (27.6(+8.4)(-7.4)+/-4.2)x10(-6), and (11.3(+3.3)(-2.9)+/-1. 4)x10(-6), respectively. Branching fraction upper limits are set for all of the other decay modes investigated. PMID- 11005964 TI - Chiral three-nucleon forces from p-wave pion production. AB - Production of p-wave pions in nucleon-nucleon collisions is studied according to an improved power counting that embodies the constraints of chiral symmetry. Contributions from the first two nonvanishing orders are calculated. We find reasonable convergence and agreement with data for a spin-triplet cross section in pp-->pppi(0), with no free parameters. Agreement with existing data for a spin singlet cross section in pp-->pnpi(+) constrains a short-range operator shown recently to contribute significantly to the three-nucleon potential. PMID- 11005965 TI - Shape of the 8B alpha and neutrino spectra. AB - The beta-delayed alpha spectrum from the decay of 8B has been measured with a setup that minimized systematic uncertainties that affected previous measurements. Consequently the deduced neutrino spectrum presents much smaller uncertainties than the previous recommendation [J. N. Bahcall et al., Phys. Rev. C 54, 411 (1996)]. The 8B nu spectrum is found to be harder than previously recommended with about (10-20)% more neutrinos at energies between 12-14 MeV. The integrated cross sections of the 37Cl, 71Ga, 40Ar, and Super-Kamiokande detectors are, respectively, 3.6%, 1.4%, 5.7%, and 2.1% larger than previously thought. PMID- 11005966 TI - Giant resonances in the doubly magic nucleus 48Ca from the (e, e'n) reaction. AB - The 48Ca(e,e(')n) reaction has been investigated for excitation energies 11-25 MeV and momentum transfers 0.22-0.43 fm(-1) at the superconducting Darmstadt electron linear accelerator S-DALINAC. Electric dipole and quadrupole plus monopole strength distributions are extracted from a multipole decomposition of the spectra. Their fragmented structure is described by microscopic calculations allowing for coupling of the basic particle-hole excitations to more complex configurations. Comparison of the excitation spectrum of the residual nucleus 47Ca with statistical model calculations reveals a 39(5)% contribution of direct decay to the damping of the giant dipole resonance. PMID- 11005963 TI - Transverse asymmetry AT' from the quasielastic 3He(e,e') process and the neutron magnetic form factor. AB - We have measured the transverse asymmetry A(T') in 3He(e,e(')) quasielastic scattering in Hall A at Jefferson Laboratory with high precision for Q2 values from 0.1 to 0.6 (GeV/c)(2). The neutron magnetic form factor G(n)(M) was extracted based on Faddeev calculations for Q2 = 0.1 and 0.2 (GeV/c)(2) with an experimental uncertainty of less than 2%. PMID- 11005962 TI - Three-pion interferometry results from central Pb+Pb collisions at 158A GeV/c. AB - Three-particle correlations have been measured for identified pi(-) from central 158A GeV Pb+Pb collisions by the WA98 experiment at CERN. A substantial contribution of the genuine three-body correlation has been found as expected for a mainly chaotic and symmetric source. PMID- 11005967 TI - Spectroscopy of double-beta and inverse-beta decays from 100Mo for neutrinos. AB - Spectroscopic studies of two beta rays from 100Mo are shown to be of potential interest for investigating both the Majorana nu mass by neutrinoless double beta decay (0nubetabeta) and low energy solar nu's by inverse beta decay. With a multiton 100Mo detector, coincidence studies of correlated betabeta from 0nubetabeta, together with the large Q value ( Q(betabeta)), permit identification of the nu-mass term with a sensitivity of approximately 0.03 eV. Correlation studies of the inverse beta decay and the successive beta decay of 100Tc, together with the large capture rates for low energy solar nu's, make it possible to detect, in real time, individual low energy solar nu in the same detector. PMID- 11005968 TI - Surface plasmon dynamics in silver nanoparticles studied by femtosecond time resolved photoemission. AB - Multiphoton photoelectron spectroscopy reveals the multiple excitation of the surface plasmon in silver nanoparticles on graphite. Resonant excitation of the surface plasmon with 400 nm femtosecond radiation allows one to distinguish between photoemission from the nanoparticles and the substrate. Two different previously unobserved decay channels of the collective excitation have been identified, namely, decay into one or several single-particle excitations. PMID- 11005969 TI - Better schemes for quantum interrogation in lossy experiments. AB - We show that several schemes exist for improving the efficiency of quantum interrogations (or "interaction-free" measurements), which in principle are limited by lossy experimental components. We show how one new scheme (the "Helstrom" scheme) can efficiently detect the presence of a bomb which has been designed to foil standard quantum interrogations by arming itself when there is high probability of a photon being in the region. This particular new scheme can probe a region of space with a greater probability of success than a person in that region determining they are being probed. PMID- 11005970 TI - Experimental real-time phase synchronization of a paced chaotic plasma discharge. AB - Experimental phase synchronization of chaos in a plasma discharge is studied using a phase variable lift technique (i.e., phase points separated by 2pi are not considered as the same). Real-time observation of synchronized and unsynchronized states is made possible through a real-time sampling procedure. Parameter space regions of synchronization and unsynchronization are identified, and a set of equations is suggested to model the real plasma circuit. PMID- 11005971 TI - Negative refractive index in left-handed materials. AB - The real part of the refractive index n(omega) of a nearly transparent and passive medium is usually taken to have only positive values. Through an analysis of a current source radiating into a 1D "left-handed" material (LHM)-where the permittivity and permeability are simultaneously less than zero-we determine the analytic structure of n(omega), demonstrating frequency regions where the sign of Re[n(omega)] must, in fact, be negative. The regime of negative index, made relevant by a recent demonstration of an effective LHM, leads to unusual electromagnetic wave propagation and merits further exploration. PMID- 11005972 TI - Pulsating, creeping, and erupting solitons in dissipative systems. AB - We present three novel pulsating solutions of the cubic-quintic complex Ginzburg Landau equation. They describe some complicated pulsating behavior of solitons in dissipative systems. We study their main features and the regions of parameter space where they exist. PMID- 11005973 TI - Resonant formation and control of 2D symmetric vortex waves. AB - It is shown that m-fold symmetric vortex waves in two dimensions ( V states) preserve their functional form in a weak straining flow having appropriate symmetry, but arbitrary time dependence. This phenomenon is used in driving the V states into a highly nonlinear excitation by subjecting a circular vortex patch to rotation and strain with oscillating strain rate and varying the rotation angular velocity. The effect is due to autoresonance in the system as the excited vortex state boundary self-adjusts its aspect ratio to synchronize with the external flow. PMID- 11005974 TI - Intense high-energy proton beams from Petawatt-laser irradiation of solids. AB - An intense collimated beam of high-energy protons is emitted normal to the rear surface of thin solid targets irradiated at 1 PW power and peak intensity 3x10(20) W cm(-2). Up to 48 J ( 12%) of the laser energy is transferred to 2x10(13) protons of energy >10 MeV. The energy spectrum exhibits a sharp high energy cutoff as high as 58 MeV on the axis of the beam which decreases in energy with increasing off axis angle. Proton induced nuclear processes have been observed and used to characterize the beam. PMID- 11005975 TI - Stabilization of neoclassical tearing modes by an externally applied static helical field. AB - The effect of a static helical magnetic field on the nonlinear growth of the neoclassical tearing mode (NTM) is investigated. The NTM is found to be stabilized by an externally applied helical field of a different helicity if the field magnitude is sufficiently large, suggesting a very simple method for stabilizing the NTM. The mechanism responsible for this stabilization is the decreased fundamental harmonic pressure perturbation of the NTM in the presence of the helical field. PMID- 11005976 TI - Visualizing interfacial structure at non-common-atom heterojunctions with cross sectional scanning tunneling microscopy. AB - We describe how cross-sectional scanning tunneling microscopy (STM) may be used to image the interfacial bonding across the nearly lattice-matched, non-common atom GaSb/InAs heterojunction with atomic-scale precision. The method, which takes advantage of the length difference between interfacial and bulk bonds, appears equally applicable to AlSb/InAs and suggests how one might recover the complete structure of either heterojunction from atomic-resolution STM data. PMID- 11005977 TI - Shear instabilities of freely standing thermotropic smectic- A films. AB - In this Letter we discuss theoretically the instabilities of thermotropic freely standing smectic- A films under shear flow [3]. We show that, in Couette geometry, the centrifugal force pushes the liquid crystal toward the outer boundary and induces smectic layer dilation close to the outer boundary. Under strong shear, this effect induces a layer buckling instability. The critical shear rate is proportional to 1/sqrt[d], where d is the thickness of the film. PMID- 11005978 TI - Pressure induces major changes in the nature of Americium's 5f electrons. AB - Americium occupies a pivotal position in the actinide series with regard to the behavior of 5f electrons. High-pressure techniques together with synchrotron radiation have been used to determine the structural behavior up to 100 GPa. We have resolved earlier controversial findings regarding americium and find that our experimental results are in discord with recent theoretical predictions. We have two new findings: (1) that there exists a critical, new structural link between americium under pressure and its near neighbor, plutonium; and (2) that the 5f electron delocalization in americium occurs in two rather than one step. PMID- 11005960 TI - Measurement of J/psi and psi(2S) polarization in pp collisions at sqrt[s] = 1.8 TeV. AB - We have measured the polarization of J/psi and psi(2S) mesons produced in pp collisions at sqrt[s] = 1.8 TeV, using data collected at the Collider Detector at Fermilab during 1992-1995. The polarization of promptly produced J/psi [psi(2S)] mesons is isolated from those produced in B-hadron decay, and measured over the kinematic range 4 [5.5] axis. The 1794-cm(-1) line is assigned to bond center H in the positive charge state, whereas the 745-cm(-1) line is ascribed to negatively charged H located on a <111> axis close to the tetrahedral site. PMID- 11005980 TI - Effect of inelastic deformation on crystallite size in post-shock 6H polytype SiC. AB - Shock-recovery experiments have been conducted on 6H polytype silicon carbide to 135 GPa and post-shock samples have been analyzed by Raman spectroscopy, x-ray diffraction, and electron microscopy. Results indicate that at up to 25-30 GPa, crystallite size is reduced faster than above this pressure, when reduction slows. Shock-induced inelastic deformation of 6H-SiC is known to develop fully for stress above 25 GPa, coinciding with the pressure range observed where crystallite size reduction decreases. Inelastic deformation appears to be responsible for this slowed crystallite size reduction. PMID- 11005981 TI - Four regimes of decaying grid turbulence in a finite channel. AB - Attenuation of second sound in helium II has been used to observe up to 6 orders of magnitude of decaying vorticity displaying four distinctly different regimes of decaying grid turbulence in a finite channel. A purely classical spectral model for homogeneous and isotropic turbulence describes most of the decay of helium II vorticity in the temperature range 1.2/=31 (chi(2) trend = 10.1). Breast-feeding for more than 1 year was associated with an OR of 0.5 (95% CI, 0.4-0.8). Forty-two (9.5%) cases and 164 (18.9%) controls reported ever oral contraceptive use: in comparison with never users, the multivariate OR was 0.4 (95% CI 0.3-0.6) for ever users, and the risk decreased with duration of use. The OR for ovarian cancer was 2.9 (95% CI, 1.5-5.8) for women with a family history of the disease. CONCLUSION: This study, conducted on a relatively low-risk population, confirms the role of oral contraceptive on ovarian cancer risk and the direct association with family history of ovarian cancer. It also indicates that a later age at first birth is directly, and induced abortion and breast feeding are inversely, related to the risk of the disease. PMID- 11006031 TI - Retrograde seeding of malignant cells during hysteroscopy in presumed early endometrial cancer. AB - OBJECTIVE: The aim of this study was to determine the effect that preoperative hysteroscopy has on the frequency of positive cytology at the time of definitive surgical management in endometrial cancer. METHODS: Charts of 222 patients with endometrial cancer were reviewed. Patients were divided according to whether (n = 64) or not (n = 158) they had hysteroscopy with saline infusion. Each group was stratified into low or high risk for positive peritoneal cytology. Logistic regression analysis was used to compare the prevalence of positive peritoneal cytology with and without hysteroscopy, before and after the stratification, adjusting for the confounding risk factors. RESULTS: After adjusting for confounding variables there was a statistical difference in the frequency of positive peritoneal cytology in those who had hysteroscopy versus those who did not (odds ratio (OR) = 2.6, 95% confidence interval (CI) = 1.02-6.63, P = 0.05). Even after stratifying patients into a low-risk group (OR = 2.12, 95% CI = 0.13 35.9, P = 0.6) and a high-risk group (OR = 3.46, 95% CI = 1.3-9.12, P = 0.01) the difference in the high-risk group was statistically significant. CONCLUSION: Hysteroscopy seems to affect the prevalence of positive peritoneal cytology, especially in those patients with high-risk cell types. Its use in patients with suspicion of endometrial cancer should be reconsidered. PMID- 11006032 TI - Combined laparoscopic and vaginal radical surgery in cervical cancer. AB - OBJECTIVE: The purpose of our study was to review our experience with laparoscopic staging and vaginal radical surgery in the treatment of early stage cervical cancer. STUDY DESIGN: We reviewed the charts of 102 patients who had a laparoscopic pelvic lymphadenectomy followed by vaginal radical hysterectomy (VRH) or vaginal radical trachelectomy (VRT). RESULTS: Patients' age ranged from 25 to 68 years (median: 36). Squamous and adenocarcinoma histology occurred in 68 and 32%, respectively. Stage Ib1 occurred in 77% of cases and the rest were stage Ia1 (1%), 1a2 (16%), and IIa (6%). Patients were divided into three groups: VRH (57), VRT (34), and node only (NO) (11), when positive nodes were identified on frozen section. Median operative time for VRH and VRT were 270 and 260 min compared to 200 min in the NO group (half also had bilateral paraaortic node dissection, which lengthened the OR time). Hospital stay was shorter in the NO group (2 days). For each group (VRH, VRT, and NO) the median pelvic node count was 27, 26, and 23 and the median paraaortic node count was 3, 4, and 9. Two VRH were converted to an abdominal procedure because of technical difficulties and one VRT was converted to a VRH because of positive endocervical margins. Intraoperative complications related to laparoscopy included two iliac and one epigastric vessel injuries. Complications related to the radical surgeries included three cystostomies, managed vaginally, and a laparotomy for parametrial bleeding after VRT. Postoperative complications occurred in 6% of patients and only one was considered major (an abscess which required surgical drainage). Overall, there were only four recurrences in the vaginal surgery groups and one in the NO group. There were no ureteral or intestinal injuries and there have been no trocar site recurrences. CONCLUSION: Our data show that approaching cervical cancer with a combined laparoscopic and vaginal surgery is feasible. The overall morbidity and complication rate are low and the lymph node count is satisfactory. Staging the nodes laparoscopically first to identify positive nodes is advantageous, particularly since we favor the use of chemoradiation therapy in those cases. The laparoscopic node staging thus avoids an unnecessary laparotomy in patients with positive nodes, reduces morbidity, and allows for early radiation therapy. PMID- 11006033 TI - Cisplatin and pentoxifylline in advanced or recurrent squamous cell carcinoma of the cervix: a phase II trial of the Gynecologic Oncology Group. AB - OBJECTIVE: The aim of this study was to determine the antitumor activity and toxicity of cisplatin and pentoxifylline in previously treated patients with squamous cell carcinoma of the cervix. METHODS: A Gynecologic Oncology Group (GOG) Phase II trial of recurrent squamous cell cervical cancer using standard GOG response and toxicity criteria was performed. RESULTS: A total of 47 patients with advanced or recurrent squamous cell carcinoma of the cervix were entered. The starting dose was 75 mg/m(2) of cisplatin every 21 days and 1600 mg of pentoxifylline PO every 8 h for nine doses during each course. Forty patients were evaluable for response and 44 were evaluable for toxicity. Of the 40 evaluable patients, 37 had received prior radiotherapy and 35 had received prior chemotherapy. A median of three courses were given (range: 1-7). Among evaluable patients, 1 had a complete response (2.5%) and 3 had a partial response (7.5%) for an overall objective response rate of 10%. The complete responder had not previously had chemotherapy. Grade 3 or 4 toxicity was predominantly nausea and vomiting (32%) and hematologic toxicity (23%). CONCLUSIONS: The combination of cisplatin and pentoxifylline at the dose and schedule tested has limited activity in previously treated advanced or recurrent cervical cancer. PMID- 11006034 TI - Constitutive activation of stat 3 oncogene product in human ovarian carcinoma cells. AB - OBJECTIVE: Stat 3 functions in transducing signals from the cell's surface to its nucleus and activation of gene transcription. Aberrations of Stat 3 in breast cancer have raised the possibility of its contribution to oncogenesis. Our goal was to examine ovarian cancer cell lines to determine whether Stat 3 plays a relevant role in ovarian carcinogenesis. METHODS: Protein lysates were extracted from normal ovarian surface epithelial cells and malignant cells. Western blotting techniques were performed with phosphorylation-independent or phosphorylation-specific Stat 3 (tyrosine 705) antibody. Confirmation of Stat 3 activation was determined by a luciferase reporter driven by a promoter containing Stat 3-specific binding sites. Bcl-x(L) and cyclin D(1) were also analyzed by Western blotting. RESULTS: MDAH 2774, OV-1063, Caov-3, and O.C. 22819 expressed high levels of phosphorylated Stat 3. In contrast, A2780 and normal ovarian surface epithelial cells had little Stat 3 phosphorylation recognized. Confirmation of persistent activation of Stat 3 activity was shown by transfection of cells with a Stat 3 luciferase reporter. Potential downstream mediators of Stat 3 including Bcl-x(L) and cyclin D(1) were also evaluated. In cells expressing activated Stat 3, high levels of both Bcl-x(L) and cyclin D(1) were detected, whereas in A2780 cells, which did not express activated Stat 3, only low levels of Bcl-x(L) and cyclin D(1) were expressed. CONCLUSIONS: Constitutive activation of Stat 3 is present in ovarian cancer lines but not in normal ovarian surface epithelial cells. Activation of Stat 3 is a common event during oncogenic transformation upstream to both Bcl-x(L) and cyclin D(1). The relationship of this aberrancy of ovarian carcinoma harboring activated Stat 3 deserves further investigation. PMID- 11006035 TI - Patients with double primary tumors in the breast and ovary- clinical characteristics and BRCA1-2 mutations status. AB - OBJECTIVE: The aim of this study was to define the prevalence, clinical characteristics, and BRCA1-2 mutation carrier status of ovarian cancer (OvC) patients with a previous primary malignancy in the breast (PPMBr). METHODS: The study population comprised 1240 consecutive Jewish Israeli women with pathologically confirmed epithelial OvC diagnosed between March 1, 1994, and December 31, 1997. Demographic and clinical data were obtained from medical files and from a detailed questionnaire taken through a nationwide epidemiological case control study on OvC. Blood samples and tumor tissues were collected for analysis of the three predominant germline BRCA1-2 Jewish founder mutations (185delAG, 5382insC, and 6174delT). RESULTS: Fifty nine (4.7%) patients with OvC had a PPMBr. The median age at diagnosis of OvC was 60 years. The mean interval between the two diagnoses was 104 months (range 0-363 months). In the majority of the patients (n = 53), the diagnosis of breast cancer (BrC) preceded the OvC by more than 1 year. The ovarian tumors were diagnosed in 47% of the cases following investigation of patients' symptoms. In 41%, diagnosis was made as a consequence of check-up exams performed during the routine follow-up of BrC survivors. Patients with PPMBr were more likely to present with FIGO ovarian stage III-IV, compared to women with solitary OvC (73% vs 60. 3%, P < 0.05), and less likely to have borderline tumors (3.4% vs 17. 9%, P = 0.007). Family history of OvC/BrC was recorded in 26% of this group of patients compared to 10.5% among patients with solitary OvC (P = 0.003). Patients with PPMBr had an exceptionally high prevalence of BRCA1-2 mutations (57%), irrespective of family history. CONCLUSIONS: Patients with PPMBr present with more advanced disease and invasive type epithelial ovarian tumors when compared to cases associated with solitary OvC. The rate of BRCA1-2 mutations in Jewish women with OvC who had PPMBr is at least twice as high as in Jewish women with OvC as the solitary disease. PMID- 11006036 TI - Age as a prognostic factor for recurrence in patients with endometrial carcinoma. AB - PURPOSE: The aim of this study was to evaluate age as a prognostic factor for recurrence in endometrial cancer patients treated with primary surgery. METHODS: Between 1983 and 1998, 455 endometrial cancer patients underwent primary surgery at our institution. Patients were divided into three age groups based on age at diagnosis: Group A (age <60, n = 156), B (age 60-69, n = 147), and C (age >/=70, n = 152). Clinicopathologic, treatment factors, and outcome were compared among the three groups. Prognostic factors were evaluated by univariate and multivariate analysis. RESULTS: The three age groups had a similar distribution of most pathologic features including stage, histology, cervical involvement, positive cytology, adnexal involvement, nodal metastases, serosal involvement, and lymphovascular invasion (LVI). Older women had a higher rate, however, of deep (>1/2) myometrial invasion (P < 0.0001) and grade 3 tumors (P < 0.0001). The extent of surgical staging and use of adjuvant radiation therapy were similar. Five-year disease-free survivals (DFS) of Groups A, B, and C were 74.3, 70.2, and 60.3%, respectively (P = 0.08). A significant difference in DFS was seen when Groups A and B were combined and compared with Group C (72.0 vs 60.3%, P = 0.03). Multivariate analysis confirmed the significance of race, stage, grade, and LVI. Age was not found to be associated with recurrence (HR 1.1, 95% C.I. 0.91-1.5, P = 0.21). CONCLUSION: Our results reveal that, in a large cohort of comparably staged and treated endometrial carcinoma patients, age is not a prognostic factor for recurrence. PMID- 11006037 TI - Age-specific survival of women with endometrioid adenocarcinoma of the uterus. AB - OBJECTIVE: The purpose of this paper was to evaluate the age-specific survival for women diagnosed with endometrioid adenocarcinoma of the uterus. METHODS: A retrospective analysis was conducted of 328 patients diagnosed with endometrioid adenocarcinoma of the uterus between January 1990 and December 1997. Patients were followed for 3 to 96 months with a mean of 43 months. The impact of age on survival was assessed using Cox proportional hazard regression and multivariate analysis for age, stage, and grade. Stage and grade were analyzed using log-rank tests, and survival curves were generated by the Kaplan-Meier method. RESULTS: A total of 328 patients were evaluated. Multivariate analysis revealed age, stage, and grade were all significant independent predictors of survival (P < 0.0001). Age-specific survival varied from a high of 90% at age 40 to a low of 55% at age 80. Interval age-specific survival decreased below 86% at age 50. Subset analysis of patients younger than 50 compared with older patients revealed no difference in surgical stage or grade of tumors among these patients. Patients older than 50, however, were 41% more likely to receive adjuvant radiation therapy. CONCLUSION: Age is a specific, significant predictor of outcome in endometrioid adenocarcinoma of the uterus. Survival decreases significantly in patients older than 50. This decreased survival associated with age is unrelated to surgical stage or grade of adenocarcinoma. Decreased survival could involve molecular differences in the developing endometrial cancer or an increased risk of death from other non-cancer-related factors. PMID- 11006038 TI - Chromosome 4 deletions are frequent in invasive cervical cancer and differ between histologic variants. AB - OBJECTIVE: Patterns of discontinuous deletion of chromosome 4 have been described in histologic variants of lung carcinomas and may represent different "hotspot" targets for gene-environment interactions. Since similar environmental risks exist for cervical cancer, we investigated patterns of discontinuous deletion in two major histologic variants. METHODS: Thirteen archival cases of squamous cell cancer (SCCA) and 11 cases of adenocarcinoma (AC) were precisely microdissected. Matched normal and tumor DNA were used for polymerase chain reaction (PCR) based loss of heterozygosity (LOH) analyses using 19 polymorphic markers spanning chromosome 4. Human papillomavirus (HPV) detection was determined by PCR using general and type-specific primers (HPV 16, 18). Differences in LOH between histologic tumor types and chromosomal regions were determined using Fisher's exact test. RESULTS: Loss at any chromosome 4 locus occurred in 92% of all tumors studied, with the majority of deletions occurring on the long arm of the chromosome. Four discrete minimal regions of discontinuous deletion (R) were identified. For these regions, LOH frequencies were 76% (R1, 4q34-q35), 48% (R2, 4q25-q26), 36% (R3, 4p15.1-p15.3), and 26% (R4, 4p16). Loss in SCCA predominated at 4q (4q34-q35; 83%) and in AC at 4p (4p15.3; 50%). Overall LOH on the p arm was significant in AC (82%) compared to SCCA (31%) (P = 0.02). HPV detection was similar in SCCA (85%) and AC (73%), and HPV 16/18 subtypes were similarly represented in both histologies. CONCLUSIONS: Chromosome 4 deletions are frequent in cervical carcinomas. Different patterns of deletion between SCCA and AC may represent gene regions targeted by different gene-environment interactions in these tumor subtypes. PMID- 11006039 TI - Cisplatin-paclitaxel-cyclophosphamide with G-CSF in primary advanced epithelial ovarian cancer. AB - INTRODUCTION: In an attempt to increase survival, we performed a prospective trial of high-dose cisplatin-paclitaxel-cyclophosphamide with granulocyte colony stimulating factor (G-CSF) for three cycles followed by carboplatin-paclitaxel for three cycles after cytoreduction of primary advanced epithelial ovarian cancer. MATERIALS AND METHODS: Thirty consecutive women with Stage 3 or 4 invasive primary epithelial ovarian cancer were treated with cytoreductive surgery. Postoperatively patients received 100 mg/m(2) of cisplatin, 200 mg/m(2) of paclitaxel, and 500 mg/m(2) of cyclophosphamide IV q 21 days x 3 cycles with 300 microg of G-CSF daily x5 beginning the first day following chemotherapy. This was followed by carboplatin AUC-5 and 135 mg/m(2) of paclitaxel IV q 21 days x3. All administration was outpatient and paclitaxel was administered over 3 h. RESULTS: Eighty percent of tumors were Stage 3C, 77% were serous, and 60% were Grade 3. Maximum cytoreduction to <2 cm was performed in 96%. Median follow-up is 30 months. Sixty-three percent of patients developed recurrence. Currently 50% of patients are alive with no evidence of disease. Estimated mean survival is 61 months and estimated mean progression-free survival is 29 months. No patient developed thrombocytopenia, neutropenic sepsis, significant neuropathy, or renal toxicity. CONCLUSION: This treatment regimen resulted in minimal toxicity and, following aggressive cytoreduction, produced good progression-free and overall survival. PMID- 11006040 TI - Absence of PTEN repeat tract mutation in endometrial cancers with microsatellite instability. AB - OBJECTIVE: PTEN, a tumor suppressor gene shown to be frequently mutated in endometrial cancers, has been suggested to be a target of microsatellite instability (MSI)-driven mutagenesis. We set out to investigate the relationship between MSI and PTEN mutation in a large series of primary endometrial carcinomas. METHODS: Thirty-nine MSI-positive endometrial cancers were evaluated by single-strand conformational variant analysis and direct sequencing to screen all nine PTEN exons for mutation. RESULTS: Fifteen specimens (38%) demonstrated 16 PTEN mutations. We observed only one alteration in the poly-adenine repeat of exon 8 that is suggested to be a target for mutation in endometrial cancers with MSI. Seven of 16 (44%) mutations in our series were deletions of >/=3 bp, a class of mutation not usually associated with tumors with defective DNA mismatch repair. To determine the significance of this high frequency of deletion, 26 additional endometrial cancers without MSI were matched with the 39 MSI-positive cancers for the prognostic factors of tumor histology, stage, grade, and patient race. The MSI-positive tumors had a significantly higher frequency of deletions involving >/=3 bp when compared with the MSI-negative group (5/11 versus 0/10, P = 0.035). CONCLUSIONS: Repeat tract mutation in PTEN is an uncommon event in MSI positive cancers. Deletion of >/=3 bp in this gene is more common in MSI-positive cancers when compared with tumors without MSI. PMID- 11006041 TI - Cervical cancer outcomes analysis: impact of age, race, and comorbid illness on hospitalizations for invasive carcinoma of the cervix. AB - OBJECTIVE: The aim of this study was to evaluate the association of age, race, and comorbid illness with procedures and complications in hospitalized patients with invasive carcinoma of the cervix in a statewide population-based database over a 3-year period. METHODS: Hospitalizations were classified into homogeneous subgroups based on a diagnosis of invasive cervical cancer. Cancer-related complications and comorbid diseases were evaluated. chi(2) and t tests determined differences in means or proportions. Linear regression techniques were applied to build models for hospitalization charges and lengths of stay (LOS). RESULTS: There were 1009 admissions. The mean age was 49.5, with a median age of 46 (21 100, SD 15.4). Of the total, 606/1009 (60%) were white, 354/1009 (35%) were African-American (AA), and 5% were "other" races. AAs were more likely to have Medicaid or be uninsured (44% vs 23%, P = 0. 001) and were more likely to be admitted for an emergency (unadjusted odds ratio (OR) = 1.6; 1.2-2.2), to have a comorbid illness (P = 0.001), to be admitted for a cancer-related complication (P = 0.036), to be admitted for a transfusion (P = 0. 01), and to be admitted for radiation therapy rather than surgery (P = 0.001). The following were associated with LOS and higher hospital costs: emergency admissions for complications of cancer, comorbid illness, and older age. CONCLUSIONS: Racial differences exist in patterns of admission, type of therapy, and severity of illness; however, there were no differences in charges or LOS for similar procedures. The large percentage of African-Americans uninsured or insured by government-supported programs indicates the potential impact of public policy on the care of these patients. Socioeconomic status rather than phenotypic appearance may be a more important determinant of outcome. PMID- 11006042 TI - Phase 2 evaluation of topotecan administered on a 3-day schedule in the treatment of platinum- and paclitaxel-refractory ovarian cancer. AB - PURPOSE; The aim of this study was to investigate the toxicity and efficacy of a more convenient topotecan administration schedule (in contrast to the "standard" 1.5 mg/m(2)/day x 5 days q 21 days) in the management of platinum- and paclitaxel refractory ovarian cancer. METHODS: Patients with clinically defined platinum- and paclitaxel-refractory ovarian cancer participating in this phase 2 trial conducted by the Gynecologic Cancer Program of the Cleveland Clinic Taussig Cancer Center received topotecan at a dose of 1.5 mg/m(2)/day x 3 days on a 21 day schedule. Both dose escalations and reductions were permitted in the protocol design. RESULTS: A total of 29 patients (median age: 61; range: 43-80) were treated with this modified topotecan schedule. These individuals had received a median of two prior regimens (range: 1-4) (retreatment with a platinum agent or paclitaxel considered a single regimen). The median number of topotecan courses delivered was 3 (range: 1-7). Major toxicity included grade 4 neutropenia (24% of patients); neutropenic fever (10%); grade 3 thrombocytopenia (10%); and requirement for blood transfusion (14%). Dose escalation was possible, and dose reductions required, in 14 and 28% of patients, respectively. Two patients exhibited evidence of a clinically relevant response to treatment. CONCLUSION: This 3-day topotecan program is more convenient and less toxic than the standard 5-day regimen. The limited level of activity observed is not inconsistent with that previously reported for the 5-day topotecan infusion schedule in platinum/paclitaxel-refractory ovarian cancer. Further investigation will be required to document the clinical utility of a 3-day topotecan schedule in a less heavily pretreated and more chemosensitive patient population. PMID- 11006043 TI - Low-grade endometrial stromal sarcoma of the ectocervix after therapy for breast cancer. AB - OBJECTIVE: Low-grade endometrial stromal sarcoma is an uncommon, indolent uterine sarcoma that can arise in extrauterine locations. The objective of this study was to report on a previously unpublished site of origin for a low-grade endometrial stromal sarcoma. METHODS: A case of a low-grade endometrial stromal sarcoma arising in the ectocervix after goserelin hormonal therapy for breast cancer was studied. RESULTS: Low-grade endometrial stromal sarcoma can arise in the ectocervix even in the absence of endometriosis. CONCLUSION: Low-grade endometrial stromal sarcoma should be included in the differential diagnosis of sarcomas of the ectocervix. PMID- 11006044 TI - Desmoplastic small round cell tumor with primary ovarian involvement: case report and review. AB - BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare, aggressive, malignant neoplasm that has recently been characterized. It has not been associated with a primary visceral organ. In women, cases are even more rare and often have some ovarian involvement. CASE: An 11-year-old girl presented with abdominal pain, nausea, and vomiting. A CT scan revealed a large heterogeneous pelvic mass with cystic components and an 8-cm midabdominal mass. During exploratory laparotomy, the patient was found to have a pelvic mass measuring 12. 9 cm replacing normal ovarian tissue. The midabdominal mass was also removed. Pathology, cytology, and immunohistochemistry confirmed a desmoplastic small round cell tumor. Even with aggressive surgical and medical intervention, the patient died 11 months after initial diagnosis. CONCLUSION: We present a rare small cell tumor that is associated with ovarian involvement. The prognosis in these patients is extremely poor and very few survivals have been reported. PMID- 11006045 TI - Endometrial carcinoma remaining after term pregnancy following conservative treatment with medroxyprogesterone acetate. AB - BACKGROUND: Successful pregnancies after conservative progestin treatment to young women with endometrial carcinoma have recently been reported. However, it is not known for certain whether the lesion is completely eradicated in such patients. We present a case of residual endometrial carcinoma after term pregnancy which had been treated conservatively before the pregnancy began. CASE: A 28-year-old woman with endometrial carcinoma received conservative treatment with high-dose medroxyprogesterone acetate (MPA) and then conceived. After delivery at term, atypical cells were found in the endometrial curettage specimen. A hysterectomy was performed 6 months after delivery and revealed the presence of a small focus of intramucosal, grade 1, endometrioid-type adenocarcinoma. Immunohistochemically, the tumor cells were positive for estrogen and progesterone receptors. CONCLUSION: We concluded that while MPA treatment had been effective, it had not completely eradicated the carcinomatous lesion, which remained during and after the term pregnancy. PMID- 11006046 TI - Cortical bone distribution in the femoral neck of strepsirhine primates. AB - The thickness of the inferior and superior cortices of the femoral neck was measured on X-rays of 181 strepsirhine primate femora representing 24 species. Neck length, neck depth and neck-shaft angle were also measured. The strength of the femoral neck in frontal bending was estimated by modeling the neck as a hollow cylinder, with neck depth as the outer diameter and cortical thickness representing the superior and inferior shell dimensions. Results indicate that the inferior cortex is always thicker than the superior cortex. The ratio of superior to inferior cortical thickness is highly variable but distinguishes two of the three locomotor groups in the sample. Vertical clingers and leapers have higher ratios (i.e., a more even distribution of cortical bone) than quadrupeds. The slow climbers tend to have the lowest ratios, although they do not differ significantly from the leapers and quadrupeds. These results do not confirm prior theoretical expectations and reported data for anthropoid primates that link greater asymmetry of the cortical shell to more stereotypical hip excursions. The ratio of superior to inferior cortical thickness is unrelated to body mass, femoral neck length, and neck-shaft angle, calling into question whether the short neck of strepsirhine primates acts as a cantilever beam in bending. On the other hand, the estimated section moduli are highly correlated with body mass and neck length, a correlation that is driven primarily by body mass. In conclusion, we believe that an alternative interpretation to the cantilever beam model is needed to explain the asymmetry in bone distribution in the femoral neck, at least in strepsirhine primates (e.g., a thicker inferior cortex is required to reinforce the strongly curved inferior surface). As in prior studies of cross sectional geometry of long bones, we found slightly positive allometry of cortical dimensions with body mass. PMID- 11006047 TI - Mass change, environmental variability and female fertility in wild Propithecus verreauxi. AB - Accurate estimates of mass and size are important in a wide range of research questions in population and evolutionary biology, and yet such data are still rare for wild primates. This study presents detailed longitudinal data from a large population of wild indriids, and demonstrates links between fluctuations in body mass, environmental cycles, and reproduction. Understanding these links is a necessary step toward explaining the function and evolution of distinctive features of lemur biology and behavior. During the first 12 years of an ongoing study of the sifaka, Propithecus verreauxi verreauxi, at Beza Mahafaly in southwest Madagascar, 320 animals were captured and weighed throughout the year. Adult males and females exhibit seasonal cycles of mass loss, with females losing significantly more mass than males. In 2 drought years this pattern was especially pronounced. Compared to lighter females, females who were heavier at the time of the mating season were more likely to give birth in the following birth season. By showing (1) seasonally greater mass loss in reproductive females compared to males, particularly in drought years, (2) a close link between female mass and fertility, and (3) an uncoupling of the periods of highest body mass and of gestation and lactation, these results suggest that energy acquisition and storage are critically important in the life history strategies of female sifaka, and that "capital breeding" may be a feature of sifaka reproductive strategies. PMID- 11006048 TI - The Iberomaurusian enigma: north African progenitor or dead end? AB - Data obtained during an ongoing dental investigation of African populations address two long-standing, hotly debated questions. First, was there genetic continuity between Late Pleistocene Iberomaurusians and later northwest Africans (e.g., Capsians, Berbers, Guanche)? Second, were skeletally-robust Iberomaurusians and northeast African Nubians variants of the same population? Iberomaurusians from Taforalt in Morocco and Afalou-Bou-Rhummel in Algeria, Nubians from Jebel Sahaba in Sudan, post-Pleistocene Capsians from Algeria and Tunisia, and a series of other samples were statistically compared using 29 discrete dental traits to help estimate diachronic local and regional affinities. Results revealed: (1) a relationship between the Iberomaurusians, particularly those from Taforalt, and later Maghreb and other North African samples, and (2) a divergence among contemporaneous Iberomaurusians and Nubian samples. Thus, some measure of long-term population continuity in the Maghreb and surrounding region is supported, whereas greater North African population heterogenity during the Late Pleistocene is implied. PMID- 11006049 TI - Paleosols, stable carbon isotopes, and paleoenvironmental interpretation of Kanapoi, Northern Kenya. AB - This study uses the interpretation of paleosol features at Kanapoi, Kenya (4.2 3.4 Ma) to reconstruct the ecosystem occupied by Australopithecus anamensis. The paleosols at Kanapoi provide a unique and fortuitous opportunity, in that the bulk of the hominid specimens derive from paleosols, providing direct evidence of the environment that the Kanapoi hominids occupied. Seven named types of paleosols are recognized at Kanapoi, each representing a trace fossil of the local ecosystem during soil formation. The hominid-bearing Dite paleosols provide evidence that A. anamensis inhabited areas of semi-arid, seasonal climate regimes with mean annual precipitation ranging from about 350-600 mm. The in situ hominid collections from Dite paleosols show that A. anamensis at least occasionally occupied relatively open low tree-shrub savanna vegetation formed in well drained settings, and may have preferred these conditions over other poorly drained soils. The relatively open conditions of Dite paleosols existed within a spatially variable ecosystem, characterized by a mosaic of environments, ranging from forb-dominated edaphic grassland to gallery woodland, providing a larger view of the mixed ecosystem in which A. anamensis lived. Synthesis of paleoenvironmental indicators of A. anamensis at Kanapoi and Allia Bay, Kenya suggests that as early as 4 Ma hominids thrived in varied ecosystems. PMID- 11006050 TI - A corrected reconstruction and interpretation of the Homo erectus calvaria from Ceprano, Italy. PMID- 11006051 TI - A re-appraisal of Ceprano calvaria affinities with Homo erectus, after the new reconstruction. PMID- 11006053 TI - Pesticides. A message from Preventive Medicine and your physician. PMID- 11006052 TI - Tobacco use among Massachusetts youth: is tobacco control working? AB - BACKGROUND: This paper examines whether the Massachusetts Tobacco Control Program is affecting the rates of smoking and smokeless tobacco use among Massachusetts' youth. METHODS: School survey data from the Massachusetts Prevalence Study were analyzed to estimate differences between 1993 and 1996 rates of youth cigarette and smokeless tobacco use, attitudes toward smoking, and awareness of cigarette ads and promotions of antismoking messages. RESULTS: Lifetime and Current Smoking rates declined significantly among middle school males, contrasting with stable national trends. Among girls in this age group, Lifetime and Current Smoking did not change significantly. Hispanic middle school students exhibited a significant decline in Lifetime Use. There were no significant changes in Lifetime or Current Smoking rates among high school students. Lifetime use of smokeless tobacco declined among middle school students while Current Use declined among both middle and high school students. Students reported declines in awareness of cigarette ads or promotions and increases in awareness of antismoking messages. CONCLUSIONS: These results provide evidence for cautious optimism regarding the impact of tobacco control, but indicate that these efforts should begin earlier and that additional research is needed to understand and address the problems of tobacco use by girls. PMID- 11006054 TI - Smokers with multiple behavioral risk factors: how are they different? AB - OBJECTIVE: The first aim of the present investigation was to examine cross sectional differences between smokers who engage in additional health risk behaviors (i.e., high-fat diets and low physical activity levels) and those who do not that could affect readiness for smoking cessation treatment and treatment prognosis. The second aim was to examine prospective associations between risk factor status and smoking outcomes (i.e., cessation and quit attempts). DESIGN: Data were derived from baseline and 1-year follow-up surveys for the SUCCESS project, a randomized trial of worksite smoking interventions conducted in 24 worksites in Minneapolis/St. Paul, Minnesota. Included in the analyses were 2,149 study participants who reported smoking at baseline. METHODS: Current smokers were categorized into one of three "risk groups": the "1 additional risk factor" group (i.e., either low physical activity level or high dietary fat intake), the "2 additional risk factor" group (i.e., both low physical activity and high dietary fat intake), and the "smoker only" group (i.e., neither low physical activity nor high dietary fat intake). Mixed model regression analyses examined cross-sectional associations between risk group status and baseline demographic variables, smoking dependency, social environments for smoking, and health problems. Prospective associations between baseline risk group status and 1-year follow-up cessation attempts and quits were also examined. RESULTS: At baseline, risk factor status was associated with smoking dependency for both men and women. Women smokers with at least one additional risk factor reported a greater number of cigarettes smoked per day, higher Fagerstrom Nicotine Dependence scores, and lower self-efficacy for refraining from smoking in a variety of situations compared with smokers with no additional risk factors. Men smokers with at least one additional risk factor reported higher Fagerstrom Nicotine Dependence scores compared with smokers with no additional risk factors. Women smokers with at least one additional risk factor were more likely to report being encouraged to quit by co-workers compared with smokers with no other risk factors. No relationship between risk factor status and social pressure to quit was observed among men. Prospective analyses indicated that baseline risk factor status was marginally related to smoking outcome at 1-year follow-up; however, these relationships were attenuated considerably when controlling for smoking dependence. Relationships between risk factor status and smoking outcomes were stronger for men. CONCLUSION: Results indicated that the presence of multiple health risk behaviors was related to more serious problems with smoking. However, the presence of additional risk factors did not strongly affect prognosis for smoking cessation. PMID- 11006055 TI - Hypocholesterolemic effect of soymilk supplementation with usual diet in premenopausal normolipidemic Japanese women. AB - BACKGROUND: The hypocholesterolemic effect of soy protein concentrates on normolipidemic subjects still remains unclear. Our objective is to assess the effect of soymilk supplementation, a whole soy product, with usual diet on serum lipids in normolipidemic subjects. METHODS: We conducted a randomized controlled trial on 60 premenopausal normolipidemic Japanese women. After excluding 8 subjects whose initial serum concentration of total cholesterol or triacylglycerol was higher than 220 mg/dL (5.69 mmol/L) or 160 mg/dL (1.81 mmol/L), respectively, we encouraged the subjects in the soymilk-supplemented group (n = 27) to consume 400 mL (408 g) of commercial regular soymilk daily during two menstrual cycles. There were no significant differences in variables, including nutrient intake, between the soymilk-supplemented and control (n = 25) groups before the intervention. RESULTS: After the trial, we observed a significant decrease of 10.9 mg/dL, or 5.3%, in serum concentration of total cholesterol in the soymilk-supplemented group. During the intervention, nutrient intake assessment showed significant increases in nutrient densities of vitamin E, polyunsaturated fatty acids, isoflavones, and P/S ratio and decreases in total energy and nutrient densities of vitamin C and green tea in this group. A statistically significant decrease in serum total cholesterol could still be observed even after excluding the estimated hypocholesterolemic effects of soymilk's polyunsaturated fatty acids. CONCLUSIONS: Our results suggest the hypocholesterolemic effect of soymilk, a traditional whole soy product, in Asian countries in normolipidemic subjects. PMID- 11006056 TI - Patient reminder letters to promote annual mammograms: a randomized controlled trial. AB - BACKGROUND: This study assessed the effects of a reminder letter from a physician (relative to a mammography facility letter or no letter) on appointment compliance among women 50-74 years of age due for an annual screening mammogram. METHODS: A total of 1,562 women were randomly as signed to the groups. Each Group 1 subject received a reminder letter from her physician, each Group 2 subject received a reminder letter from her mammography facility, and Group 3 served as a control group. RESULTS: The return rates for Groups 1, 2, and 3 were 47.7, 46.6, and 28.3%, respectively; the overall difference was significant using a chi(2) analysis (P < 0.001). Bonferroni pairwise comparisons indicated no difference between Groups 1 and 2 but significant differences (P < 0.001) between Group 3 and the other two groups. Logistic regression indicated that relative to Group 3, the adjusted odds of returning for Groups 1 and 2 were 2.37 and 2.24, respectively. CONCLUSIONS: Mammography providers and their patients likely will benefit from in-reach reminder systems. Physicians who do not use reminder systems should refer their patients to facilities that use these systems. PMID- 11006057 TI - Psychosocial influences on older adults' interest in participating in bowel cancer screening. AB - BACKGROUND: As part of a multicenter, randomized controlled trial of the efficacy of flexible sigmoidoscopy for the prevention of bowel cancer, an investigation of the predictors of screening interest was carried out in a subsample of older adults. METHOD: The aim of the study was to establish the predictive power of the Health Belief Model (HBM) and to evaluate the contribution of HBM elements in mediating the effect of other demographic and health variables which have been found to be associated with screening interest and participation. A total of 5,099 participants were sent a postal questionnaire which examined screening interest, attitudes toward screen ing (benefits and barriers), perceived bowel cancer risk, bowel cancer worry, bowel symptoms, health status, state anxiety, and optimism. A total of 3,648 questionnaires were returned completed, giving a response rate of 71.5%. RESULTS: The results showed that threat, barriers, and benefits explained 47% of the variance in interest. Demographic and health variables were also associated with screening interest, although most of their effect was mediated by the HBM constructs. DISCUSSION: This community study in older adults showed a high level of interest in participating in screening. The large sample size provided the opportunity to test the value of the HBM model and to examine mediation of demographic and health variables. The HBM proved to be a good model of screening interest. These results further our understanding of the decision processes in participating in cancer screening and point to directions to increase the level of participation in community samples. PMID- 11006058 TI - Population frequency distributions of HDL, HDL(2), and HDL(3) cholesterol and apolipoproteins A-I and B in healthy men and women and associations with age, gender, hormonal status, and sex hormone use: the Stanford Five City Project. AB - BACKGROUND: The purpose of this study is to present population-based frequency distribution data for several lipoprotein-related variables and to examine their associations with gender, age, menopausal status, and sex hormone use. METHODS: High-density lipoprotein cholesterol (HDL-C), HDL(2)-C, HDL(3)-C, apolipoprotein (Apo) A-I, and Apo B were measured in a population-based sample of 1, 027 healthy adults from four California cities who participated in the 1989-1990 survey of the Stanford Five City Project. These data were examined cross-sectionally with sociodemographic and other related variables. RESULTS: Relative to men, all of the HDL-related parameters-HDL-C, HDL(2)-C, HDL(3)-C, Apo A-I-were significantly higher and Apo B levels were significantly lower among women (P < 0. 001). Menopausal status was not associated with HDL-related parameters, but Apo B levels were higher in post- versus premenopausal women (P < 0.001). Among women, HDL-C and Apo A-I levels were higher in oral contraceptive and estrogen replacement therapy users (P = 0.003). Most of the significant findings remained statistically significant after adjusting for age, body mass index, smoking, energy expenditure, and alcohol intake. CONCLUSIONS: These population-based data indicate that gender, menopausal status, and the use of sex hormones among women are associated with differential levels of one or more of HDL-C, HDL(2)-C, HDL(3) C, Apo A-I, and Apo B, independent of age and a broad set of lifestyle factors. PMID- 11006059 TI - Associations of HDL, HDL(2), and HDL(3) cholesterol and apolipoproteins A-I and B with lifestyle factors in healthy women and men: the Stanford Five City Project. AB - BACKGROUND: Measures of the two major high-density lipoprotein (HDL) subfractions, HDL(2) and HDL(3), and the major apolipoproteins of HDL and low density lipoprotein (LDL), Apo A-I and Apo B, may be etiologically important factors in the development of coronary artery disease. The association of lifestyle factors with these lipoprotein-related variables remains unclear. METHODS: HDL-C, HDL(2)-C, HDL(3)-C, Apo A-I, and Apo B levels were determined in a population-based sample of 1,027 healthy women and men aged 25-64 years, from four California cities who participated in the 1989/1990 survey of the Stanford Five City Project. In this cross-sectional study we examined the independent associations of these lipoprotein-related variables with body mass index (BMI), cigarette smoking, daily energy expenditure, alcohol intake, dietary intake, and hormone use (oral contraceptives and estrogen replacement therapy). RESULTS: In general, BMI and alcohol intake were the strongest independent predictors of the lipoprotein-related variables. The negative association of BMI with HDL-C was attributable primarily to the association with the HDL(2)-C subfraction, while for alcohol intake the positive association with HDL-C was attributable primarily to the association with HDL(3)-C, particularly in men. Among men, but not women, energy expenditure was a significant independent predictor of each of the lipoprotein-related variables, with positive associations observed for HDL-C, HDL(2)-C, HDL(2)-C, and Apo A-I and a negative association observed for Apo B (P < 0.005). CONCLUSIONS: Data from this population-based sample suggest that specific lifestyle factors are more strongly associated with some lipoprotein related variables than with others, with notable gender differences. PMID- 11006060 TI - Telephone counseling as adjuvant treatment for nicotine replacement therapy in a "real-world" setting. AB - BACKGROUND: Physicians prescribing nicotine replacement therapy (NRT), or health plans covering NRT, often want their patients to receive adjuvant behavioral treatment. However, how to do that in a "real world" is unclear. This paper reports results from a public health program that uses proactive telephone counseling as support for physician advice and provides adjuvant treatment for NRT users. METHODS: Participants were NRT users (N = 8,832) who called the California Smokers' Helpline, a statewide cessation service that provides proactive counseling, one session before NRT use and multisessions after the smokers received NRT. After receiving NRT, some participants discontinued the counseling while others continued with follow-up sessions. A subset of the 8,832 participants (n = 664) was interviewed 13 months later for quitting status. RESULTS: After receiving NRT, 79% of the participants continued with counseling and received 4.2 sessions on average, while 21% of them received only one session. Overall, 82.8% of all participants made a quit attempt. Nicotine patch users were more likely to make an attempt than nicotine gum users (85.2% vs 66.3%), but the relapse probability was the same for these attempts. Those who received multiple counseling were more likely to make an attempt than those receiving single counseling (84.4% vs 77.1%) and were more likely to stay quit for 1 year (25.6% vs 16.1%). CONCLUSIONS: Proactive telephone counseling is a promising adjuvant treatment for NRT users in a "real-world" setting: a convenient referral service for supporting health plans or physicians who advise their patients to quit smoking. PMID- 11006061 TI - Smokers ages 50+: who gets physician advice to quit? AB - BACKGROUND: Smoking-related morbidity and mortality, and benefits associated with quitting, extend across the life span. Health care provider interventions enhance quitting. The present study examined perceived influence of physician advice to quit and characteristics of subjects receiving this advice. METHODS: Subjects were 1,454 smokers ages 50+ with at least one physician visit in the past year. Subjects were surveyed at baseline for receipt of and reactions to physician advice to quit and for smoking, health, and demographic characteristics. RESULTS: Over half of subjects welcomed physician advice to quit, about half said the advice influenced their quitting decision "extremely" or "quite a lot," and about one-third indicated that it increased their confidence in quitting. Physicians were more likely to advise sicker patients, indicated by poorer health status, at least one past year hospitalization, and presence of cardiovascular, cerebrovascular, or respiratory diseases. CONCLUSIONS: Midlife and older smokers reacted generally favorably to physician advice to quit. Physicians were more likely to advise patients with commonly recognized smoking-related diseases. Discrepancies were noted in advice given to sicker vs healthier patients. Additional physician training in less commonly recognized smoking-related illnesses, intervening with healthier patients to prevent disease, and enhancing patients' confidence in quitting may improve outcomes. PMID- 11006062 TI - Cardiovascular disease risk factor intervention in low-income women: the North Carolina WISEWOMAN project. AB - OBJECTIVES: The North Carolina WISEWOMAN project was initiated to evaluate the feasibility of expanding an existing cancer screening program to include a cardiovascular disease (CVD) screening and intervention program among low-income women. METHODS: Seventeen North Carolina county health departments were designated as minimum intervention (MI), and 14 as enhanced intervention (EI). The EI included three specially constructed counseling sessions spanning 6 months using a structured assessment and intervention program tailored to lower income women. RESULTS: Of the 2,148 women screened, 40% had elevated total cholesterol (> or = 240 mg/dL), 39% had low high-density lipoprotein cholesterol (HDL-C) levels (< 45 mg/dL), and 63% were hypertensive (systolic blood pressure 140 and/or diastolic blood pressure > or = 90 mm Hg or on hypertensive medication). The majority of women (86%) had at least one of these three risk factors. Seventy six percent were either overweight or obese. After 6 months of follow-up in the EI health departments, changes in total cholesterol levels, HDL-C levels, diastolic blood pressure, and BMI were observed (-5.8 mg/dL, -0.9 mg/dL, -1.7 mm Hg, and -0.3 kg/m(2), respectively), but were not significantly different from MI health departments. A dietary score that summarized fat and cholesterol intake improved by 2.1 units in the EI group, compared with essentially no change in the MI group. CONCLUSIONS: Expanding existing cancer screening programs to include CVD intervention was feasible and may be an effective means for promoting healthful dietary practices among low-income women. PMID- 11006063 TI - A randomized trial of a tailored, self-help dietary intervention: the Puget Sound Eating Patterns study. AB - BACKGROUND: This study evaluated a tailored, multiple-component self-help intervention designed to promote lower fat and higher fruit and vegetable consumption. METHODS: Participants were 1,459 adults selected at random, stratified by sex and age (18-34, 35-54, 55-69), from enrollees of a large health maintenance organization. After completing a baseline telephone survey, participants were randomized to receive the intervention (consisting of a computer-generated personalized letter, a motivational phone call, a self-help manual, a package of supplementary materials, computer-generated behavioral feedback based on a self-administered food frequency questionnaire, and newsletters) or to receive no materials. Evaluation was based on 1,205 (86.5%) participants who completed both a 3- and a 12-month follow up survey. RESULTS: The intervention effect +/- SE for fat, based on a diet habits questionnaire, was -0.10 +/- 0.02 (P < 0.001), corresponding to a reduction of approximately 0.8 percentage points of percentage energy from fat. For fruits and vegetables, the intervention effect was 0.47 +/- 0.10 servings/day (P < 0.001). Intervention effects were similar across age and sex groups. CONCLUSIONS: Tailored, self-help interventions can effectively promote dietary change among both men and women and among younger as well as older adults. PMID- 11006064 TI - Hypercholesterolemia and 5-year risk of development of coronary heart disease among university and school workers in Jeddah, Saudi Arabia. AB - BACKGROUND: A study was performed among university and school employees as a step in detecting the prevalence of coronary heart disease (CHD) risk factors among school and university staff and the relation of the various risk factors to hypercholesterolemia and to predict the 5-years probability of development of CHD. METHODS: All university staff and a sample of school workers in Jeddah were included in the study. Data were collected by interview and self-administered questionnaires, as well as by direct observation and measurement of weight, height, blood pressure, and total blood cholesterol. Logistic regression analysis was performed to identify the impact of various risk factors on hypercholesterolemia. The 5-year risk of development of CHD was calculated to identify the proportion at highest risk. RESULTS: Some CHD risk factors were detected among the participants as 18.8% were current cigarette smokers, around half were classified as overweight, and 19.9% were classified as hypertensive. Hypercholesterolemia was present in 10.1% and was significantly related to older age (40 years and over) and place of work. The estimated risk of CHD in the coming half-decade for those classified in the fifth quintile was 0.068. The Q(5)/Q(1) ratio has shown that those classified in the fifth quintile were at 3.6 times more risk of developing CHD in the coming 5 years than those classified in the first quintile. CONCLUSION: CHD risk factors are becoming prevalent in our society. The risk of development of CHD in the coming decades is not trivial. Short- and long-term health strategies are recommended to decrease the risk of CHD and improve the quality of life. PMID- 11006065 TI - Evaluation of a population-based screening for type 2 diabetes: a community-based screening project in Puli, Taiwan. AB - BACKGROUND: A Markov method incorporating the relationships between prevalence, incidence, and mortality with respect to type 2 diabetes was used to assess a population-based screening for this disease. METHODS: Data from a population based screening project for residents of Puli, Taiwan, over 30 years of age (n= 1,219) were used to estimate the annual incidence of asymptomatic type 2 diabetes, the prevalence to incidence (P/I) ratio, and the hazard rate of death due to type 2 diabetes. These parameters were employed to develop a Markov process to evaluate the effects of early detection of type 2 diabetes on the risk of death from this disease in a simulated population (n= 10,000) receiving biennial, 5-year interval, or no screening. RESULTS: The estimated annual incidence, average duration from asymptomatic to symptomatic type 2 diabetes (P/I ratio), and hazard rate for death from this disease were 0.86% (95% CI 0.50 1.48), 10 years (95% CI 7.69-14.01), and 1.1% per year, respectively. This yields an optimal screening interval of 5 years. Simulation of a 5-year interval screening regimen versus no screen ing yielded a relative risk reduction of 31% (95% CI 12-46%). A similar value was found for a biennial screening regime. CONCLUSIONS: The results suggest that early detection of type 2 diabetes via a community-based screening project in developing countries with high prevalence is worthwhile. PMID- 11006066 TI - 7-year stability of blood pressure in the Canadian population. AB - BACKGROUND: The purpose of the study was to examine the 7-year stability of systolic (SBP) and diastolic (DBP) blood pressures in the Canadian population. METHODS: The sample included 1,503 participants 7-69 years of age from the 1981 Canada Fitness Survey who were remeasured in Campbell's Survey of 1988. Both SBP and DBP were adjusted for the effects of body mass index (BMI) using regression procedures. RESULTS: Interage correlations from baseline to follow-up ranged from -0.17 to 0.61 for SBP and from -0.22 to 0. 51 for DBP. With few exceptions, correlations were positive and significant, and were highest and most consistent in adulthood. Further, between 27 and 39% of participants in the upper or lower quintiles in 1981 remained there in 1988. There were few differences in adiposity between those who remained in the upper or lower quintiles and those who did not. One exception was that males who remained in the upper quintile of SBP had greater values for BMI, sum of skinfolds, and waist circumference at baseline. Among adults, the best predictor of future blood pressure was baseline blood pressure, which accounted for between 12 and 34% of the variance in follow-up blood pressure, followed by age, follow-up BMI, and, in females, baseline physical activity levels. CONCLUSIONS: Blood pressure demonstrated low to moderate stability over 7 years in Canada, and baseline level of adiposity was related to the stability of SBP in males. PMID- 11006067 TI - Predictors of stage of adoption for colorectal cancer screening. AB - BACKGROUND: Although colorectal cancer is the third most common cancer in women, little is known about predictors of adherence to screening. METHODS: A randomly selected sample of 202 predominantly low-income and African-American women were interviewed. Knowledge of, attitudes and beliefs about, and practices related to flexible sigmoidoscopy (FS) screening were assessed. RESULTS: The majority of participants were in the precontemplation stage of adoption (56%). There were significant differences by stage of adoption for FS beliefs, FS barriers, risk of developing colorectal cancer, worry about getting colorectal cancer, and physician recommendation to get a FS. Predictors of adherence to FS guidelines were perceiving fewer barriers to getting a FS and having a physician recommend a FS. CONCLUSION: Seventy-two percent of the women in this study were nonadherent to FS screening guidelines. Psychosocial factors play an important role in screening for colorectal cancer. Ways of reducing barriers and increasing physician recommendations should be explored. PMID- 11006068 TI - Attitudes, beliefs, and knowledge as predictors of nonattendance in a Swedish population-based mammography screening program. AB - BACKGROUND: The effectiveness of mammography screening could be improved if factors that influence nonattendance were better understood. METHODS: We examined attitudes, beliefs, and knowledge in relation to nonattendance in a population based mammography screening program, using a case-control design. Data were collected from November 1997 to March 1998 through telephone interviews with 434 nonattenders and 515 attenders identified in a population-based mammography register in central Sweden. The questions asked drew primarily upon the components constituting the Health Belief Model. RESULTS: Multivariate analysis showed that nonattendance was most common among women within the highest quartile of perceived emotional barriers, compared to women within the lowest quartile (OR = 4.81; 95% CI 2.96-7.82). Women who worried most about breast cancer were more likely to attend than those who worried least (OR = 0.09; 95% CI 0.02-0.31). Women with the highest scores of perceived benefits were more likely to attend than women with the lowest ones (OR = 0.35; 95% CI 0.08-0.75). Other factors associated with nonattendance were less knowledge about mammography and breast cancer, lack of advice from a health professional to participate, and very poor trust in health care. CONCLUSIONS: Our findings suggest that increased participation in outreach mammography screening programs can be achieved through enhancement of breast cancer awareness and possibly by reducing some of the modifiable barriers. mammography; mass screening; breast cancer; attitudes; Sweden. PMID- 11006069 TI - Effectiveness of interventions to improve follow-up after abnormal cervical cancer screening. AB - OBJECTIVE: The purpose of this study was to determine the effectiveness of interventions designed to improve follow-up after an abnormal Pap smear. METHODS: We performed a qualitative meta-analysis of interventions designed to improve follow-up after an abnormal Pap smear and included studies that met the following criteria: randomized or concurrently controlled study design, defined outcomes, and data available for abstraction. Interventions were classified as behavioral, cognitive, sociologic, or combined strategies (e.g., behavioral and cognitive). Abnormal Pap smears were defined as any test result requiring additional follow up. Effectiveness was measured by the rate of compliance with recommended follow up. RESULTS: Twenty-two interventions in 10 studies met the inclusion criteria. Cognitive interventions utilizing interactive telephone counseling were the most effective, improving compliance by 24 to 31%. Behavioral interventions, such as patient reminders, were also effective, increasing follow-up by up to 18%. Not all of these results achieved statistical significance. The single sociologic intervention we identified used video-taped peer discussions to provide a message about abnormal Pap smears and appropriate follow-up. This intervention was not associated with increased follow-up after an abnormal test. The effectiveness of interventions using multiple types of strategies to improve follow-up was inconsistent. CONCLUSIONS: Cognitive strategies led to the greatest improvement in compliance with follow-up of abnormal Pap smear screening tests. Extension of similar interventions to follow-up of abnormal breast and colon cancer screening, development of physician- and system-targeted interventions, and evaluation of the cost-effectiveness of these strategies are important priorities for future research. PMID- 11006070 TI - Feasibility of a tailored intervention to improve preventive care use in women. AB - BACKGROUND: Women age 50 years and older are in need of multiple preventive health care services. Despite recent improvements in rates of delivery of preventive care services, especially within managed care organizations, substantial numbers of women are still being underscreened. Efforts to improve delivery of preventive care services have often focused on one outstanding service despite the fact that patients often are in need of many services. METHODS: A total of 893 women age 50 to 55 years were mailed a self-administered survey to identify outstanding preventive health care service needs. Patients in need of three or more outstanding preventive health care services were identified from survey respondents to participate in a feasibility study evaluating a tailored, customized intervention called Tic Tac Health. RESULTS: Five-hundred ninety-one women returned the survey (67%). Four-hundred forty-eight (76%) women were in need of one or more preventive health services; 92 (16%) were in need of three or more. Twenty-two patients (24%) completed the Tic Tac Health card. The women who completed the card were similar to those who did not. CONCLUSIONS: Despite documented physician visits, presence of managed care health insurance, and a designated primary care provider, a significant number of women are still in need of multiple preventive health services. An intervention targeting multiple preventive health services was demonstrated to be both feasible and effective. Further evaluation via a randomized controlled trial should be conducted to determine if an intervention like Tic Tac Health would be an effective modality for improving rates of receipt of multiple preventive health care services. PMID- 11006071 TI - Cancer of the colon and rectum in California: trends in incidence by race/ethnicity, stage, and subsite. AB - BACKGROUND: Incidence and mortality from cancer of the colon and rectum have declined in recent years in the United States and California, but reasons for the decline are unknown. METHODS: Age-adjusted site-specific and stage-specific incidence rates were calculated for approximately 9,000 cases of in situ cancer and 120,000 cases of invasive cancer of the colon and rectum diagnosed between 1988 and 1996 among California residents and reported to the California Cancer Registry. Trends in incidence over time were measured using the estimated annual percent change. RESULTS: Among non-Hispanic whites there was a decline in all sites and stages, but the decrease was most pronounced for rates of in situ and regional/distant tumors in the rectum and sigmoid which declined by about 4 to 7% a year. For tumors in the proximal colon, the decrease was statistically significant only for regional/distant tumors which declined about 2% a year. Among blacks, there was an approximately 7% annual decline in the incidence of regional/distant tumors of the rectum in women and a nearly 3% a year decrease in regional/distant tumors of the proximal colon in men. The decline in rates for Hispanics and Asian/Pacific Islanders was smaller and less consistent than for non-Hispanic whites. CONCLUSIONS: The results confirm a overall decline in all stages of cancer of the colon and rectum in California, particularly among non Hispanic white men and women. The decrease was most pronounced for tumors in the rectum and sigmoid colon and may be attributable to screening. PMID- 11006072 TI - The role of regucalcin in nuclear regulation of regenerating liver. AB - Regucalcin was discovered in 1978 as a Ca(2+)-binding protein that does not contain EF-hand motif of Ca(2+)-binding domain [Yamaguchi, M., and Yamamoto T., Chem. Pharm. Bull. 26, 1915-1918, 1978]. The name regucalcin was proposed for this Ca(2+)-binding protein, which can regulate liver cell functions related to Ca(2+). Regucalcin has been demonstrated to play a multifunctional role in liver and kidney cells, for which regucalcin mRNA expression and its protein content are pronounced. Hepatic regucalcin mRNA expression has been shown to be mediated through signaling pathway of Ca(2+)/calmodulin-dependent protein kinase, protein kinase C, and tyrosine kinase. AP-1- and NF-1-like factors can bind to the promotor region of the rat regucalcin gene to mediate the Ca(2+) response for transcriptional activation. Growing evidence supports the view, moreover, that regucalcin plays an important role in the regulation of Ca(2+) signaling from the cytoplasm to nuclei in the proliferative cells of regenerating rat liver. Also, regucalcin has been demonstrated to be transported to liver nucleus, and it can inhibit nuclear protein kinase, protein phosphatase, and DNA and RNA synthesis in regenerating liver. Regucalcin plays a physiologic role in the control for overexpression of proliferative cells. Regucalcin has been proposed to be an important regulatory protein in nuclear signaling system. PMID- 11006073 TI - Synthesis of stereoisomeric analogues of endomorphin-2, H-Tyr-Pro-Phe-Phe-NH(2), and examination of their opioid receptor binding activities and solution conformation. AB - All sixteen stereoisomeric analogues of endomorphin-2 (H-Tyr-Pro-Phe-Phe-NH(2)) were synthesized by Fmoc-strategy using solid phase methods. Although synthetic endomorphin-2 exhibited similar mu- and delta-opioid receptor-binding activity to the natural compound, endomorphin-2 analogues containing d-amino acid isomers exhibited lower interaction with mu-receptors depending on the particular combination. The data clearly indicated that the three dimensional structure of endomorphin-2 with the natural l-configuration was the most suitable for binding within the mu receptor, but specific residues are important for activity. Circular dichroism studies verified that changes in chirality of amino acids in the endomorphin-2 sequence resulted in structural conformation. These alterations significantly reduced the specificity for mu-receptor-binding sites. PMID- 11006074 TI - A novel gene (retinovin) expressed selectively in the early stage of chick retinal development. AB - To understand molecular mechanisms of retinal development, genes expressed selectively only in the early stage of retinal development were isolated by subtractive hybridization based on suppression polymerase chain reaction. The retina has no layered structure in 7-day chick embryos, in contrast with the fully developed multilayered structure of neurons in 15-day embryos. The subtraction between cDNA derived from retinal tissues at these different stages, followed by repeat rounds of 5'-RACE (rapid amplification of cDNA ends) and 3' RACE, led to isolation of a novel gene with an open reading frame encoding a putative protein with 753 amino acids. Its specific expression in the 7-day embryonic retina was confirmed by Northern blot analysis. The gene, named "retinovin," would be used as a marker for identifying retinal stem cells present at the early stage of retinal development. PMID- 11006075 TI - AMP-Activated protein kinase is activated by the stimulations of G(q)-coupled receptors. AB - The AMP-activated protein kinase (AMPK) functions as a metabolic sensor that monitors cellular AMP and ATP levels. Platelet-activating factor (PAF) activates endogeneous AMPKalpha1 in Chinese hamster ovary cells expressing the PAF receptor coupled with both G(i) and G(q), but its activity was not inhibited after treatment with islet-activating protein. Norepinephrine and bradykinin also activated AMPKalpha1 in cells expressing the G(q)-coupled alpha(1b)-adrenergic receptor and bradykinin receptor, respectively. Stimulations of the G(i)-coupled alpha(2A)-adrenergic receptor, fMet-Leu-Phe receptor, prostaglandin EP3alpha receptor, and G(s)-coupled beta(2)-adrenergic receptor did not activate AMPKalpha1. AMPKalpha1 thus is activated specifically by stimulation of G(q) coupled receptors. G(q)-coupled receptors transmit the signal for GLUT4 translocation and glucose uptake through an insulin-independent pathway. However, direct activation of AMPKalpha1 with treatment of 5-aminoimidazole-4-carboxamide 1-beta-d-ribofuranoside did not trigger GLUT4 translocation nor stimulate glucose uptake in our cells. Thus, activation of AMPKalpha1 via G(q) is not sufficient to trigger GLUT4 translocation or stimulate glucose uptake. PMID- 11006076 TI - Complete amino acid sequence of Japanese chestnut agglutinin. AB - The complete amino acid sequence of Japanese chestnut (Castanea crenata Sieb. et Zucc.) agglutinin (CCA) was determined. Analysis by SIMS of the acidic peptide obtained by pepsin digestion revealed that the N-terminal amino acid sequence should be Acetyl-Met-Glu-Glu. Prior to sequence analysis, redetermination of cysteine residues indicated the presence of one cysteine residue per subunit. The complete sequence was determined by endoproteinase Arg-C and Achromobacter protease I digestion, and CNBr cleavage. CCA consists of 309 amino acid residues with a high content of glycine (16.5 mol%) and one cysteine residue. The calculated molecular mass was 33, 387 Da including the N-terminal acetyl group. C terminal sequence analysis of intact CCA gave only one sequence, HMEYF, indicating that no heterogeneous CCA formed by posttranslational cleavage at the C-terminal region, as occurs in some legume lectins. Analysis of the sequence of CCA itself revealed that CCA could be divided into two structural domains, the N domain and the C-domain, almost at the center. These domains share about 35% identical residues, so CCA has a repeat sequence. Also, both domains show a homology to jacalin-related lectins with 27-38% identity. These results suggest that the structure of CCA resembles two molecules of jacalin-related lectin. PMID- 11006077 TI - Edema induction by the disintegrin-like/cysteine-rich domains from a Bothrops atrox hemorrhagin. AB - Viperine and crotaline snake venoms contain one or more hemorrhagic metalloproteases called hemorrhagins. The most potent hemorrhagins belong to the P-III class and have, in addition to the protease domain, disintegrin-like and cysteine-rich domains. Although proteolytic degradation of vascular endothelium basement membrane has been established to be the main factor responsible for hemorrhage, several studies reveal other factors that actually do facilitate this process. Recent evidence has shown that the nonprotease domains of the P-III class hemorrhagins are able to inhibit the platelet aggregation by blocking essential procoagulant integrins on platelets. In this study we report the identification of a hemorrhagin from Bothrops atrox venom. This enzyme, a P-III class metalloprotease, undergoes an apparent spontaneous degradation, releasing a proteic fragment containing the disintegrin-like/cysteine-rich domains. This fragment shows the capability to induce an edematogenic process, suggesting the existence of a still unknown nonenzymatic mechanism of vascular permeability increase. PMID- 11006078 TI - ATP synthesis and heat production during Ca(2+) efflux by sarcoplasmic reticulum Ca(2+)-ATPase. AB - Vesicles derived from the sarcoplasmic reticulum of rabbit white skeletal muscle were loaded with Ca(2+) and used to measure the rates of Ca(2+) efflux, heat production, and ATP synthesis from ADP and P(i). It was found that the Ca(2+) ATPase can function in three different forms: (i) it absorbs heat from the medium (5 Kcal/mol Ca(2+)) when the efflux was coupled with ATP synthesis; (ii) it converts the energy derived from the gradient into heat (30 Kcal/mol Ca(2+)) when Mg(2+) is removed from the medium and the synthesis of ATP is impaired; and (iii) the ATPase becomes uncoupled when the different ligands of the enzyme are removed from the medium. As a result, there is no ATP synthesis and no heat production or absorption during Ca(2+) efflux. The Ca(2+) efflux, ATP synthesis, and heat production were inhibited by thapsigargin, a specific inhibitor of the Ca(2+) ATPase. PMID- 11006079 TI - Suppression of CFTR-mediated Cl(-) secretion by enhanced expression of epithelial Na(+) channels in mouse endometrial epithelium. AB - The present study investigated the effect of enhanced expression of epithelial Na(+) channels (ENaC) on the cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion in the mouse endometrium using the short-circuit current technique. The amiloride sensitivity of the basal current of the cultured endometrial epithelia was found to vary with the magnitude of the basal current, the higher the basal current the greater its sensitivity to amiloride, indicating possible elevation of ENaC expression. However, the magnitude of the forskolin induced Isc, previously demonstrated to be mediated by CFTR, decreased as the amiloride sensitivity of the basal current increased, suggesting a possible inhibitory effect of elevated expression of ENaC on CFTR-mediated Cl(-) secretion. The Matrigel concentration for culturing the endometrial epithelia was found to affect the amiloride sensitivity of the basal current as well as the forskolin-induced Isc in opposite directions. However, competitive RT-PCR demonstrated that the expression of both ENaC and CFTR was enhanced in Matrigel treated culture, suggesting that the reduced forskolin-induced Isc with enhanced amiloride sensitivity was not due to a reduction in CFTR expression, but rather suppression of CFTR function by enhanced ENaC expression. In addition to the previously demonstrated inhibition of ENaC by activation of CFTR, the present results reveal possible regulation of CFTR by ENaC. The interaction between the two may be one of the underlying mechanisms for balancing Na(+) absorption and Cl(-) secretion across epithelia. PMID- 11006080 TI - Molecular cloning of testis-abundant finger Protein/Ring finger protein 23 (RNF23), a novel RING-B box-coiled coil-B30.2 protein on the class I region of the human MHC. AB - We have identified a genomic DNA fragment, using the PCR method with degenerate oligonucleotide primers which contain the conserved sequence of the RING finger domain. Using the DNA fragment as a probe, a novel cDNA was cloned from human and mouse testis. The cDNA had a domain structure of the typical RING-B box-coiled coil (RBCC)-B30.2 domain and therefore was named testis-abundant finger protein (tfp). Indeed, the transcript was highly expressed in the testis, although it was also found ubiquitously in various organs by Northern blot analysis. The tfp gene was mapped at the class I region of the human MHC (major histocompatibility complex), within which some known RBCC-B30.2 proteins such as RFP, RFB30/HERF1, AFP, and HZF had been localized. These findings demonstrate that several RBCC B30.2 proteins including tfp, which are non-HLA proteins, are clustered within the class I region of the human MHC. PMID- 11006081 TI - Multiple transcripts of the human Cu,Zn superoxide dismutase gene. AB - We have identified five alternatively spliced transcripts of the gene for human Cu,Zn superoxide dismutase (SOD1), a causative gene for autosomal dominant amyotrophic lateral sclerosis (ALS). The splice variants of wild-type or mutant SOD1 were expressed in a tissue-specific manner; therefore, their expression may be regulated to modify SOD1 function. In addition, the expression in the brain implies that variants may play a role in the nervous system, the region involved in ALS. Immunoblot study of HeLa cells transfected with two variants encoding C terminal truncated proteins did not show the proteins of expected size. However, this observation is consistent with the previous study of C-terminal truncated mutant proteins that cause ALS, suggesting that both variant and mutant proteins may share certain properties, such as instability or insolubility in the cytosol. These findings suggest that the splice variants may contribute to a physiological function of SOD1 or to a pathological mechanism in ALS. PMID- 11006082 TI - Inorganic Polyphosphate/ATP-NAD kinase of Micrococcus flavus and Mycobacterium tuberculosis H37Rv. AB - An enzyme with both inorganic polyphosphate [poly(P)]- and ATP-dependent NAD kinase activities was isolated from Micrococcus flavus. The enzyme was a dimer consisting of 34 kDa subunits, and was named poly(P)/ATP-NAD kinase. Internal amino acid sequences of the enzyme showed homologies with some function-unknown proteins released on the GenBank database. Among such proteins, hypothetical Rv1695 protein (Accession No. Z98268-16), which was encoded by a gene named "Rv1695" on genomic DNA of Mycobacterium tuberculosis H37Rv, was proposed to be poly(P)-dependent NAD kinase. By cloning and expression in Escherichia coli, Rv1695 was shown to encode poly(P)/ATP-NAD kinase and named ppnk. The ppnk product, recombinant-poly(P)/ATP-NAD kinase (Ppnk) was purified and characterized. The enzyme was a tetramaer consisting of 35 kDa subunits when expressed in E. coli. Poly(P)/ATP-NAD kinases of M. flavus and Ppnk of M. tuberculosis H37Rv specifically and completely phosphorylated NAD by utilizing commercially available poly(P)s and nucleoside triphosphates as phosphoryl donors. PMID- 11006083 TI - ppGpp-dependent leuO expression in bacteria under stress. AB - Despite the known potential transcription regulatory role of leuO gene product, LeuO, the condition when leuO expresses during bacterial growth cycle remains unclear. Mechanistically, leuO expression was shown to be part of promoter relay mechanism, however, the factor(s) responsible for the regulation of leuO expression is not known. Combining Northern and Western results, we demonstrate in the present communication that leuO expression is normally low and enhanced when bacteria are in transition from exponential growth to stationary phase. The stationary phase-associated leuO expression is ppGpp dependent and rpoS (sigma(s) factor) independent. PMID- 11006084 TI - Enzymes leading to the nucleotide sugar precursors for exopolysaccharide synthesis in Burkholderia cepacia. AB - Based on the chemical composition of the exopolysaccharide produced by the cystic fibrosis bacterial isolate Burkholderia cepacia IST408, we postulated and confirmed, based on the specificity of enzymes detected in crude cell-free extracts, the pathway leading to the presumptive activated sugar precursors: UDP D-glucose, UDP-D-galactose, UDP-D-glucuronic acid, GDP-D-mannose, and GDP-D rhamnose. Results also suggest that regulation of the expression of the mucoid phenotype in B. cepacia does not occur at the level of synthesis of any of these enzymes. PMID- 11006085 TI - Induction of NAD(+)-linked 15-hydroxyprostaglandin dehydrogenase expression by androgens in human prostate cancer cells. AB - Prostate cancer cells are known to express cyclooxygenases (COXs) and synthesize prostaglandins. Catabolism of prostaglandins in these cells remains to be determined. Induction of NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH), a key metabolic inactivation enzyme, was investigated in androgen sensitive LNCaP cells and in hormone-independent PC3 cells. 15-PGDH was found to be induced by dihydrotestosterone or testosterone in a time- and dose-dependent manner in LNCaP but not in PC3 cells as shown by activity assay and immunoblot analysis. However, prostaglandin synthetic enzymes, COX-1 and COX-2, were not found to be induced by androgens. Induction was also achieved by 17beta-estradiol and progesterone, although to a lesser extent. Induction of 15-PGDH was not blocked by steroid receptor antagonist, RU 486, nor by antiandrogen, flutamide. However, induction was inhibited by tyrosine kinase inhibitor, genistein, and by ERK kinase inhibitor, PD 98059, but not by protein kinase C inhibitor, GF109203X. These results suggest that androgens induce 15-PGDH gene expression through an unconventional nongenomic pathway. PMID- 11006086 TI - Effects of carvedilol on isolated heart mitochondria: evidence for a protonophoretic mechanism. AB - Carvedilol (?1-[carbazolyl-(4)-oxy]-3-[2-methoxyphenoxyethyl)amino]-pro panol-(2) ?) is a novel compound used in clinical practice for the treatment of congestive heart failure, mild to moderate hypertension, and myocardial infarction. Carvedilol was also shown to protect cardiac mitochondria from oxidative stress events. Because mitochondria are the main suppliers of ATP for cardiac muscle work, a study of the effects of carvedilol in mitochondrial bioenergetics is necessary to fully understand the basis of its protective role in myocardial energetics. In this work we show that carvedilol acts as an uncoupler of oxidative phosphorylation, decreasing mitochondrial electric potential (DeltaPsi) by a weak protonophoretic mechanism. Theoretical studies were carried out to determine the relevance of conformation and proton affinity of the protonable amino side-chain group in the proton-shuttling activity across the inner mitochondrial membrane. BM910228, a hydroxylated metabolite of carvedilol, was also studied for comparison with the parent compound. Implications for the protective role of carvedilol in heart mitochondrial bioenergetics are discussed. PMID- 11006088 TI - Expression of the Na(+)/Ca(2+) exchanger ameliorates ionomycin-induced cell death. AB - PC12 cells were stably transfected with cDNA encoding the Na(+)/Ca(2+) exchanger (NCX1.4). A robust Na(+)-dependent Ca(2+) uptake confirmed the functional expression of the protein. When NCX1. 4 expressing cells (NO) and vector transfected control cells (VC) were exposed to 0.5-20 microM ionomycin for 6 h, a dose-dependent increase in LDH release was observed. LDH release was significantly reduced in NO when compared with VC. When either VC and NO were treated with 3 microM ionomycin and 1.1 mM EGTA, the increase in LDH release was nearly abolished. However, when VC and NO were treated with ionomycin and then EGTA was added 2 min later, LDH release remained elevated. These data suggest ionomycin-induced cell death was Ca(2+) dependent and expressing NCX1.4 may have ameliorated cell death by reducing elevated [Ca(2+)](I). PMID- 11006087 TI - Pyrrolidine dithiocarbamate inhibits TNF-alpha-dependent activation of NF-kappaB by increasing intracellular copper level in human aortic smooth muscle cells. AB - Pyrrolidine dithiocarbamate (PDTC) is a metal-chelating compound that acts as antioxidant or pro-oxidant and is widely used to study redox regulation of cell function. In the present study, we investigated effects of PDTC and another antioxidant, N-acetyl-l-cysteine (NAC), on TNF-alpha-dependent activation of NF kappaB in human aortic smooth muscle cells (HASMC). Treatment of the cells with TNF-alpha induced the activation of p65/p50 heterodimer NF-kappaB and increased the mRNA levels of monocyte chemoattractant protein (MCP)-1. Pretreatment with PDTC markedly suppressed the NF-kappaB activation and expression of MCP-1 by inhibiting IkappaB-alpha degradation. In contrast, NAC had no effect. PDTC concomitantly increased the intracellular levels of copper, and bathocuproinedisulfonic acid, a non-cell-permeable chelator of Cu(1+), inhibited the PDTC-induced increase in intracellular copper level and reversed the PDTC effects on IkappaB-alpha, NF-kappaB, and MCP-1. These results indicate that TNF alpha-dependent expression of MCP-1 in HASMC is tightly regulated by NF-kappaB and that intracellular copper level is crucial for the TNF-alpha-dependent activation of NF-kappaB in HASMC. PMID- 11006090 TI - Three-hybrid strategy reveals a peptide segment that specifically binds to the 3' untranslated region of NF-IL6 mRNA. AB - The 3'UTR of eukaryotic mRNA is an important regulation region, on which many trans factors act. In recent years, a series of 3'UTRs were shown to have tumor suppressor function, including the 3'UTR of the human nuclear factor for interleukin-6 (NF-IL6 3'UTR). To understand molecular basis for this function, we have tried to isolate genes encoding protein factors acting on the RNA of NF-IL6 3'UTR. Here we show that, by using a yeast three-hybrid system, a cDNA fragment was successfully isolated. This cDNA was allowed to express in E. coli, and its expression product, a polypeptide of ca. 70 amino acids long, was shown to specifically bind to the NF-IL6 3'UTR RNA. A search in GenBank did not reveal homologous sequences. Therefore, this cDNA fragment may be a part of the gene of a novel NF-IL6 3'UTR specific binding protein. PMID- 11006089 TI - Calcium-modulating cyclophilin ligand desensitizes hormone-evoked calcium release. AB - The Ca(2+)-modulating cyclophilin ligand (CAML) protein causes stimulation of transcription factors via activation of a store-operated Ca(2+) entry pathway. Since CAML is widely expressed in mammalian tissues, it may be an important regulator of Ca(2+) store function. In the present study, we investigated the consequence of CAML overexpression on Ca(2+) signaling using rapid confocal imaging of Fluo3-loaded NIH3T3 fibroblasts. Control and CAML-expressing cells gave concentration-dependent responses to the Ca(2+) mobilizing agonist ATP. CAML expression reduced the sensitivity of the cells so that higher concentrations of ATP were needed to achieve global Ca(2+) waves. The amplitudes of Ca(2+) waves were significantly reduced in CAML expressing cells, consistent with earlier suggestions that CAML causes depletion of internal Ca(2+) stores. With low ATP concentrations, only local Ca(2+) release events were observed. CAML did not affect the characteristics of these local Ca(2+) signals, suggesting that it does not directly affect Ca(2+) release channels. PMID- 11006091 TI - Alternative splicing forms of the human CD1D gene in mononuclear cells. AB - CD1d is a critical molecule for the presentation of lipid antigens to natural killer (NK) T cells. To investigate the molecular complexity of CD1d, alternatively spliced transcripts in peripheral blood mononuclear cells from three healthy subjects were analyzed by PCR and sequencing methods. We found eight alternatively spliced variants of the CD1D gene (V1-V8), seven of which are newly established variants (V2-V8). V1 and V4 are in-frame; however, the other six variants (V2, V3, V5-V8) are out-of-frame. V1, V2, V4, and V5 lack a beta(2) microglobulin binding site (alpha3 domain), indicating the unstable presentation of the CD1d molecule on the surface. In V2 and V5, the transmembrane region is absent, supporting a soluble CD1d. In the V3-V8 variants, the antigen binding region (alpha1 and alpha2 domains) is partially defective, suggesting incomplete functional products. In contrast, the V1 and V2 transcripts bear the complete antigen binding site, resulting in functional proteins. Especially, the V2 splicing variant might function as an inhibitory soluble CD1d molecule and regulate the presentation of antigens on APC to NKT cells. PMID- 11006092 TI - Characterization of NAADP(+) binding in sea urchin eggs. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP(+)) is a pyridine nucleotide which has been shown to release Ca(2+) from intracellular membranes in echinoderms, Ascidiae, mammals, and plants. NAADP releases Ca(2+) via a mechanism independent of ryanodine and inositol 1,4,5-trisphosphate (IP(3)) receptors and the NAADP(+) receptor is likely to be located on a separate organelle. We have investigated the binding characteristics of NAADP(+) to its receptor in sea urchin egg homogenates. NAADP(+) binds to a saturable membrane-bound site with high affinity (K(d) = 193 +/- 35. 7 pM). NAADP(+) associates to its receptor with a t(1/2) of approximately 7 min while dissociation does not occur during the time course of the experiment. Furthermore, NAD(+), NAAD(+), ADP, or ATP cannot displace NAADP(+) binding. The structurally related molecules NADP(+) and NADPH displayed a markedly lower affinity for the binding site with K(d)'s 500- and 25,000-fold higher than NAADP(+), respectively. This discrepancy between oxidized and reduced forms of NADP(+) might suggest that NAADP(+) signaling is itself regulated by the redox state of the cell. PMID- 11006093 TI - Molecular cloning and genomic organization of the mouse AE2 anion exchanger gene. AB - The molecular organization of the AE2 (SLC4A2) gene, a member of the multigene family encoding sodium-independent chloride/bicarbonate anion exchangers, has previously been described in both humans and rats. In these two species, AE2 shows alternate promoter usages and tissue-specific expression of isoforms in a similar, but not identical, fashion. Here we report the molecular cloning and organization of the entire mouse AE2 gene. The gene consists of 23 exons and 22 introns and spans about 17 kb. Moreover, it drives transcription of N-terminal truncated isoforms from alternate promoter sequences in a way analogous to that described for rat and/or human orthologs. Thus, sequences within intron 2 function as overlapping alternate promoters for truncated isoforms AE2b(1) and AE2b(2), and sequences of intron 5 drive transcription of isoforms AE2c(1) and AE2c(2). Each of these variants has a specific alternative first exon, while remaining exons are common to the complete form of the message AE2a, the diversity at 5' leading to different N-termini in corresponding encoded proteins. As expected, mouse AE2 promoter sequences and the patterns of tissue expression of AE2 isoforms resemble rat counterparts more closely than human ones. PMID- 11006094 TI - Genomic organization of the human kallikrein gene family on chromosome 19q13.3 q13.4. AB - Kallikreins are a subgroup of serine proteases with diverse physiological functions. Recently, growing evidence indicates that many kallikrein genes are involved in malignancy. In rodents, kallikreins are encoded by a large multigene family, but in humans only three kallikreins were thought to exist. Based on the homology between the human and rodent kallikrein loci, we studied a 300 kb region of genomic sequences around the putative KLK1 gene locus on chromosome 19q13.3 q13.4. By using linear sequence information, restriction analysis, end sequencing, PCR and blotting techniques, as well as bioinformatic approaches, we were able to construct the first detailed map of the human kallikrein gene family. Comparative analysis of genes located in this area, provides strong evidence that the human kallikrein gene family locus on chromosome 19 is considerably larger than previously thought, containing at least fifteen genes. We have established, for the first time, the common structural features that apply to all members of the expanded kallikrein multigene family. Our map specifies the distance between genes to one base pair accuracy, the relative location, and the direction of transcription of all 15 genes. Determination of the true size of the kallikrein family in humans is important for our understanding of the contribution of the kallikreins to human biology and pathophysiology. PMID- 11006095 TI - Hypoxia/hypoglycemia-induced amino acid release is decreased in vitro by preconditioning. AB - The aim of this study was to investigate the effects of preconditioning on amino acid neurotransmitter release, induced by hypoxia/hypoglycaemia, from rat brain cortical slices. Tissue, perfused with artificial cerebrospinal fluid (aCSF) at 37 degrees C with zero glucose and gassed with 95% nitrogen and 5% carbon dioxide, showed a fivefold increase in glutamate release with little effect on gamma-aminobutyric acid (GABA) release. Preconditioning, with three 5-min periods of hypoxia/hypoglycaemia preceding continuous hypoxia/hypoglycaemia, significantly decreased glutamate release whilst significantly elevating GABA release. These results suggest that GABA may reduce the release of glutamate and consequently decrease the neurotoxic effects of glutamate. PMID- 11006096 TI - Identification of five new isoforms of murine thrombopoietin mRNA. AB - Thrombopoietin (Tpo) is the major physiologic regulator of platelet production. Its gene is expressed in many organs and appears constitutive in liver and kidney. However, inducible gene expression in the bone marrow and spleen have been reported as well as the presence of a number of isoforms, presumably arising from alternative splicing. We have identified five new murine Tpo isoforms, designated Tpo 5 to Tpo 9. Alternative splicing of these isoforms, in addition to the already-reported four isoforms, occurs around exon 7, the last exon, with insertion of some intron sequences or deletion of exon sequences. Studies of tissue distribution indicate that Tpo 4 is the major isoform in lymph nodes and bone marrow. The roles of these isoforms in hematopoietic regulation are unknown, but the presence of inducible Tpo mRNA in the marrow microenvironment may contribute to platelet or stem cell homeostasis. PMID- 11006097 TI - Inhibiting proteasomes in human HepG2 and LNCaP cells increases endogenous androgen receptor levels. AB - Treating HepG2 cells with MG132 for 4 h to inhibit proteasomal activity increased androgen receptor immunoreactivity in two major bands with molecular weights of 102 and 110 kDa by 77% each (P < 0. 05). MG132 treatment also increased the overall level of polyubiquitinated proteins between 66 and 220 kDa by 140% (P < 0.05). Antiubiquitin immunoreactivity comigrating with the androgen receptor bands was also increased by MG132 treatment. Two other proteasome inhibitors, lactacystin and epoxomycin, caused similar increases in the androgen receptor in HepG2 cells. Proteosome-inhibition studies conducted in LNCaP cells also showed that the two major androgen receptor bands with molecular weights of 102 and 110 kDa were increased by 85 and 115%, respectively (P < 0. 05 for both) by MG132 treatment. Overall levels of polyubiquitinated proteins with molecular weights between 66 and 220 kDa increased 365%. Ubiquitin immunoreactivity comigrating with the androgen receptor bands was also significantly increased. Thus inhibiting proteasomes in two human androgen-responsive cell lines increases endogenous androgen receptor levels as well as androgen receptor-associated ubiquitin-modified immunoreactivity. The regulation of steady-state levels of endogenous androgen receptor by proteasomal degradation could be involved in its rapid turnover in the absence of ligand and would provide a mechanism for limiting androgen responses. A PEST sequence similar to one in the vitamin D receptor is present in the hinge region of all known mammalian androgen receptors, suggesting that it may function in proteasome-mediated androgen receptor turnover. PMID- 11006099 TI - Probing the topology of the glutamate receptor GluR1 subunit using epitope-Tag insertions. AB - At least two different models for the transmembrane topology of the glutamate receptor subunits have been proposed. We investigated some features of these two models for the GluR1 subunit by inserting epitope tags between residues Lys(502) Pro(503), Ala(632)-Glu(633), Lys(712)-Pro(713), or after the C-terminal residue Leu(889). The accessibility of the tags then was detected using a tag-specific antibody before and after detergent-permeabilizing oocytes expressing the tagged subunits. The epitope tag inserted between residues Lys(712)-Pro(713) is extracellular and after Leu(889) intracellular. Epitope tags inserted between residues Lys(502)-Pro(503) and residues Ala(632)-Glu(633) were not detectable. Collectively, these results provide supporting evidence for a previously proposed topological model of GluR subunits containing an N-terminal extracellular domain, three transmembrane domains, the first two of which are bridged by a reentrant membrane pore-lining loop, and an intracellular C-terminal domain. PMID- 11006098 TI - Genistein induces apoptosis of RPE-J cells by opening mitochondrial PTP. AB - Although previous studies demonstrated that genistein-induced apoptosis of various cell types including RPE-J cells, the involvement of mitochondrial events in such types of apoptosis has not been demonstrated to date. In this investigation of genistein-induced apoptosis of RPE-J cells, genistein induced the reduction of the mitochondrial membrane potential and the release of cytochrome c to cytosol. A mitochondrial permeability transition pore (PTP) blocker bongkrekic acid prevented the reduction of the mitochondrial membrane potential and cytochrome c release, and consequently abolished caspase-3 activation, nuclear condensation, and DNA fragmentation. On the other hand, zVAD fmk did not inhibit the mitochondrial event such as the reduction of the mitochondrial membrane potential and cytochrome c release although it prevented caspase-3 activation, nuclear condensation, and DNA fragmentation. Taken together, genistein induces apoptosis of RPE-J cells by opening the mitochondrial PTP, and the mitochondrial event in this type of apoptosis is caused independently of caspase. PMID- 11006100 TI - Regulation of proteins involved in insulin signaling pathways in differentiating human adipocytes. AB - In the present study we have examined the proteins involved in the insulin signaling cascade during and after differentiation of human adipocyte precursor cells and their correlation with glucose uptake. The differentiation of human adipocytes was characterized by a two- to threefold stimulation of glucose transport in response to insulin and a marked increase protein expression for the insulin receptor, IRS-1, GLUT-4, PI 3-kinase, and PKB, with respect to undifferentiated cells. In contrast, there were small changes in the protein expression of IRS-2, and no changes in PKC zeta and MAP kinases, although basal MAP kinase activity and GLUT-1 protein were reduced during differentiation. In conclusion, there are quantitative differences in the regulation of IRS-1 and other proteins during differentiation which may contribute to more efficient insulin signaling leading to glucose uptake in mature fat cells. Alterations in this pattern may reflect or contribute to an insulin-resistant state. PMID- 11006101 TI - Wavelength-programmed solute release from photosensitive liposomes. AB - Liposomes of dipalmitoylphosphatidylcholine containing a photochromic lipid "Bis Azo PC" release entrapped solutes on exposure to UV light. We have now demonstrated that on addition of cholesterol (up to 25 mol%) to the liposomal membrane the liposomes also release their contents in response to visible light in the region of 470 nm, to which liposomes lacking steroid are insensitive. In a mixed population of liposomes prepared with and without cholesterol, this enables wavelength-dependent release of entrapped solutes on sequential exposure to visible and UV light. Furthermore, the cholesterol-containing liposomes allow stepped partial release of entrapped solute following multiple periods of short visible illumination. It is suggested that the cholesterol-containing liposomes may be potentially useful for drug delivery and for "caging" of reagents. PMID- 11006102 TI - Hepatic copper accumulation induces DNA strand breaks in the liver cells of Long Evans Cinnamon strain rats. AB - Effects of accumulation of copper and iron on the production of DNA strand breaks were investigated in Long-Evans Cinnamon (LEC) strain rats that spontaneously develop fulminant hepatitis. Copper and iron accumulated in the liver of LEC rats in an age-dependent manner from 4 to 15 weeks. Low-copper food prevented the accumulation of copper in the liver, but did not prevent accumulation of iron. When the amounts of DNA single strand breaks were estimated by comet assay, the number of DNA strand breaks in the liver cells of rats fed standard food increased with age from 4 to 15 weeks. The number of DNA strand breaks in the liver cells from rats fed low-copper food were the same as those of rats at 4 weeks of age. Thus, the copper accumulation in the liver of LEC rats induced DNA strand breaks, but accumulation of iron did not. PMID- 11006103 TI - The subcellular location of nucleoside analog phosphorylation is a determinant of synergistic effects of hydroxyurea. AB - The ribonucleotide reductase inhibitor hydroxyurea exhibits synergistic pharmacological activity with several nucleoside analogs used in antiviral and anticancer chemotherapy. We have used a cell model system where a deoxycytidine kinase (dCK)-deficient cell line was reconstituted with genetically engineered dCK targeted to the cytosol, the nucleus, or the mitochondria to investigate how the subcellular location of nucleoside analog phosphorylation affected the synergistic effects of a ribonucleotide reductase inhibitor. Hydroxyurea showed synergistic cytotoxicity with the nucleoside analogs 1-beta-d arabinofuranosylcytosine and 2-chloro-2'-deoxyadenosine when dCK was expressed in the cytosol or in the nucleus, but not when dCK was expressed in the mitochondria. These data indicate that the synergistic effect of ribonucleotide reductase inhibition is limited to nucleoside analogs phosphorylated in the cytosol or the cell nucleus. PMID- 11006104 TI - Myristoylation alters retinoic acid-induced down-regulation of MARCKS in immortalized hippocampal cells. AB - The myristoylated alanine-rich C kinase substrate (MARCKS) is a prominent PKC substrate in the brain, which has been implicated in brain development, cytoskeletal remodeling, calcium/calmodulin signaling, and neuroplasticity. The sequence of the Macs gene codes for a protein that has three highly conserved domains including a 5' myristoylation region and a 25-amino-acid phosphorylation site domain (PSD), which are involved in anchoring MARCKS to the cellular membrane. In this study, we examined the role of the myristoylation signal in the regulation of MARCKS in transfected rat hippocampal cells (H19-7) following retinoic acid (RA) treatment. A mutant MARCKS lacking the myristoylation signal was engineered by substitution of alanine for glycine at position 2 of the Macs gene and was found to be exclusively expressed in the cytosol fraction of transfected cells. Exposure of the wild-type MARCKS-transfected cells to RA resulted in an apparent shift of MARCKS from the membrane to the cytosol, while the total protein of wild-type MARCKS was not significantly changed. In contrast, RA-exposed cells transfected with the mutant MARCKS revealed a dramatic reduction of expression of MARCKS protein in both cytosol and total protein fractions. These data suggest that the absence of the myristoyl moiety may not only alter the anchoring of the protein to the membrane but also play a novel role in modulating cellular levels of MARCKS protein in response to RA. PMID- 11006106 TI - Characterization of l-asparaginase fused with a protective ScFv and the protection mechanism. AB - A fusion protein of the protective scFv linked to the C-terminus of ASNase via (Gly(4)Ser)(6) peptide was constructed. The ASNase-scFv fusion protein expressed in Escherichia coli exists mainly in the form of inclusion bodies, and a small amount of it was soluble. The soluble form was purified by four-step purification and it has been demonstrated that ASNase-scFv fusion exists as a dimer. By assay of the stability against proteolysis, the ASNase-scFv fusion was found to be more stable than native ASNase but less stable than scFv-ASNase fusion. The results of immunological assay indicated that the immunogenicity of the fusion proteins increased while their binding capacity with the anti-ASNase serum decreased by comparison to the native ASNase. Moreover, here the comparison of the basic physical and chemical properties of the ASNase-scFv fusion, scFv-ASNase fusion, and native ASNase is presented. Based on the structural evidence and the biochemical analysis described in this paper, the protection mechanism proposed in our previous study was further supported. The scFv moiety of the fusion protein may confer the ASNase moiety resistance to proteolysis as a result of both steric hindrance such as blocking the cleavage sites of trypsin and a change in the electrostatic potential surface of the enzyme. PMID- 11006105 TI - Glucocorticoid resistance in thymocytes from mice expressing a T cell receptor transgene. AB - A majority of thymocytes undergo apoptosis during differentiation due to lack of survival signals provided by T cell receptor (TCR) activation. As glucocorticoids (GC) have been suggested to be involved in this process, we have investigated the GC sensitivity in thymocytes from mice expressing a transgenic selecting TCR. We now report that immature CD4(+)CD8(+) double-positive thymocytes from these mice are comparatively more resistant to corticosterone-induced apoptosis. This is associated with reduced glucocorticoid receptor (GR) expression, increased levels of membrane CD28, increased NF-kappaB DNA binding activity, and increased binding to the CD28 response element in the interleukin-2 gene promoter. Analysis of NF kappaB/Rel proteins from nuclear extracts demonstrated altered levels of some of these proteins. Our results suggest that TCR recognition of self major histocompatibility antigens generates intracellular signals which alter the thymocyte GC sensitivity and thereby protect them against apoptosis induced by endogenous GC. PMID- 11006107 TI - Functional properties of sodium channels do not depend on the cytoskeleton integrity. AB - Several observations suggest an interaction of the sodium channel alpha-subunit with the cytoskeletal structures. However, there is a wide variability in the results of experiments of heterologous expression in Xenopus oocytes and studies on mammalian cells are sometimes contradictory. In general, there has been no direct demonstration that ad hoc large perturbations of the cytoskeleton modify the intrinsic properties of the sodium channels expressed endogenously or heterologously in plasma membranes. We have studied in CHO cells transfected with the rat muscle sodium channel alpha-subunit the effects of two substances expected to produce drastic perturbations of the cytoskeletal structure: Cytochalasin-D, which depolymerizes microfilaments, and Colchicine, which inhibits the microtubules polymerization. We observed no significant differences in the voltage dependence, kinetic parameters and surface density of the expressed sodium channels after treatment of the cells with these substances. We conclude that the two known main components of the cytoskeleton do not interfere directly with the sodium channel function or with the heterologous expression of channels in the cell membrane. PMID- 11006109 TI - Cysteine is the initial site of modification of alpha-crystallin by kynurenine. AB - Tryptophan metabolites, such as kynurenine, are spontaneously unstable at neutral pH. They undergo side-chain deamination yielding reactive alpha, beta unsaturated ketones. In the lens, where these compounds act as UV filters, reaction of the breakdown products with lens proteins (crystallins) may be largely responsible for age-dependent colouration of this tissue. In previous research, where high pH (pH 9) was used to promote deamination and conjugation with lens protein, histidine, lysine, and cysteine residues were found to be modified. In this study we show that, at pH 7, site of reaction with the major lens chaperone alpha crystallin, is the single cysteine residue of the alphaA subunit. This apparent selectivity has important ramifications because the cysteine-kynurenine adduct is itself unstable under physiological conditions. PMID- 11006108 TI - Evidence for the cluster model of mitochondrial steroid hydroxylase system derived from dissociation constants of the complex between adrenodoxin reductase and adrenodoxin. AB - Using biotinylated adrenodoxin and avidin-Sepharose 4B, dissociation constants for the complex between adrenodoxin reductase and adrenodoxin in the oxidized and reduced states were determined as 50 +/- 11 and 296 +/- 44 nM, respectively. Concentrations of adrenodoxin reductase/adrenodoxin in the matrix fraction from bovine adrenal cortex, liver, and kidney mitochondria were determined to be 20.2 +/- 10.6 microM/120 +/- 23 microM, 0.17 +/- 0.06 microM/1.79 +/- 0.24 microM, and 0.40 +/- 0.23 microM/1.33 +/- 0.26 microM, respectively. The calculation of the percentage of adrenodoxin reductase in the complex form in the reduced state showed that it is higher than 99% in adrenal cortex mitochondria, providing clear evidence for the cluster model for the mitochondrial steroid hydroxylase system. PMID- 11006110 TI - Site-directed mutagenesis of the substrate-binding cleft of human estrogen sulfotransferase. AB - The sulfonation of estrogens by human estrogen sulfotransferase (humSULT1E1) plays a vital role in controlling the active levels of these hormones in the body. To understand more fully the structural and functional characteristics of humSULT1E1, we have carried out site-directed mutagenesis of critical amino acids found in the substrate-binding cleft. Three single amino acid mutations of humSULT1E1 (V145E, H107A, and K85A) were created in this study. Kinetic studies were used to provide information about the importance of these residues in substrate specificity and catalysis, using a variety of substrates. Lysine at position 85 has been proposed to be within hydrogen bonding distance to the 3alpha-phenol group of beta-estradiol, thereby stabilising the substrate in the active site. However, substitution to a neutral alanine at this position improved substrate specificity of humSULT1E1 for beta-estradiol, estrone, and dehydroepiandrosterone (DHEA). The exchange of valine 145 for negatively charged glutamic acid markedly improved the ability of humSULT1E1 to sulfonate dopamine, but caused a reduction in specificity constants toward steroids tested, in particular DHEA. The presence of a histidine residue at position 107 was shown to be essential for the production of a functional protein, as substitution of this amino acid to alanine resulted in complete loss of activity of humSULT1E1 towards all substrates tested. PMID- 11006111 TI - Equivalent death of P-glycoprotein expressing and nonexpressing cells induced by the protein kinase C inhibitor staurosporine. AB - P-glycoprotein (P-gp) is an ATP-dependent drug pump that confers multidrug resistance. In addition to its ability to efflux toxins P-gp can also inhibit apoptosis induced by a wide array of cell death stimuli that rely on activation of intracellular caspases for full function. We have previously demonstrated that stimuli including drugs such as hexamethylene bisacetamide (HMBA), the cytotoxic lymphocyte granule protein granzyme B, and pore-forming proteins such as perforin, kill P-gp positive cells in a caspase-independent manner. We therefore hypothesised that drugs that are not effluxed by P-gp and which induce cell death in the absence of caspase activation could induce death of P-gp expressing cells. Staurosporine has been previously shown to kill cells in the absence of caspase activation. Consistent with our hypothesis, we demonstrate here that staurosporine can equivalently kill P-gp(+ve) and P-gp(-ve) tumor cell lines in a caspase-independent manner. PMID- 11006112 TI - Increase in X-ray-induced mutations by exposure to magnetic field (60 Hz, 5 mT) in NF-kappaB-inhibited cells. AB - It is established that extremely low frequency magnetic fields (ELFMF) at the flux densities, i.e., 5 mT and less, are not mutagenic. However, exposure to ELFMF enhances mutations induced by X-rays. In this study, we examined the effects of long-term exposure to 5 mT ELFMF on mutation induction and X-ray induced mutations in human malignant glioma cells (MO54) with different mutant IkappaB-alpha (a critical inhibitor of NF-kappaB) genes. Cells were exposed or sham-exposed to 5 mT ELFMF for up to 8 days with or without initial X-rays (4 Gy), and the mutant frequency of hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene was analyzed. An obvious increase in X-ray-induced mutations was observed after treatment with ELFMF in combination with X-irradiation in MO54 cells with tyrosine mutant IkappaB-alpha gene other than with serine mutant IkappaB-alpha gene or vector alone. Exposure to ELFMF alone increased mutations significantly in MO54 cells with tyrosine mutant IkappaB-alpha gene. In addition, X-ray-induced apoptoic cells were increased in MO54-V cells after exposure to ELFMF, while an anti-apoptotic effect of magnetic field was found in MO54-SY4 cells. Our data suggest that exposure to 5 mT ELFMF may induce mutations and enhance X-ray-induced mutations, resulting from the inactivation of NF-kappaB through the inhibition of tyrosine phosphorylation. PMID- 11006113 TI - Molecular cloning and chromosomal mapping of a novel five-span transmembrane protein gene, M83. AB - In an attempt to identify novel transmembrane molecules expressed on hematopoietic cells, we identified a novel transmembrane protein gene, M83. Cloning of the full-length cDNAs of human and mouse M83 revealed that M83 encodes a type I transmembrane protein with a region containing five hydrophobic segments within the C-terminal part of the protein, suggesting that M83 is a five-span transmembrane molecule. The M83 protein was expressed on the cell surface as a glycosylated protein with a molecular mass of 84 kDa. The M83 gene was localized to human chromosome 16p13.3, mouse chromosome 17B1, and rat chromosome 10q12.3 distal. In human, M83 mRNA was highly expressed in placenta, pancreas, and lymphohematopoietic tissues including peripheral blood, spleen, and bone marrow. Among hematopoietic cells, it was highly expressed in resting T lymphocytes and was downregulated by cell activation, suggestive of its biological role related to the T cell resting status. PMID- 11006114 TI - Peptide-mediated transcytosis of phage display vectors in MDCK cells. AB - Delivery of therapeutic macromolecules and gene vectors to certain tissues is hampered by endothelial or epithelial barriers. We show here that the transport of phage particles across epithelial cells can be facilitated by peptide ligands selected from a phage library of random peptides. Using MDCK cells, we identified a polycationic peptide sequence, RYRGDLGRR, containing a putative integrin binding (RGD) motif that enhanced basal-to-apical transcytosis of peptide-bearing phage 1000- to 10,000-fold compared with phage with no peptide insert. Both the synthetic peptide RYRGDLGRR and the integrin-binding peptide GRGDSP inhibited phage transcytosis suggesting the involvement of integrins. Confocal immunofluorescence microscopy showed that following internalization at the basal cell surface, phage particles were delivered to the apical cytoplasm and released at the apical cell surface. These data suggest the feasibility of using short peptides for targeting transcytotic pathways and facilitating delivery of macromolecules across cellular barriers. PMID- 11006115 TI - Cloning, functional characterization, and expression of thyrotropin receptors in the thyroid of amago salmon (Oncorhynchus rhodurus). AB - Two thyrotropin receptor cDNAs (sTSH-Ra and sTSH-Rb) were cloned from thyroid tissue of the amago salmon, Oncorhynchus rhodurus. sTSH-Ra and sTSH-Rb showed the highest degrees of sequence homology to mammalian TSH receptors. Functional characterization in COS-7 cells transiently transfected with sTSH-Ra or sTSH-Rb showed the largest increase in cAMP when exposed to bovine TSH. RT-PCR analysis demonstrated that sTSH-Ra and sTSH-Rb were expressed in the basibranchial region, but not in the ovary, testis, liver, kidney or brain. In situ hybridization revealed that sTSH-Ra and sTSH-Rb were exclusively expressed in thyroid follicular epithelial cells of amago salmon undergoing smoltification. These results indicated that the cloned cDNAs encode functional TSH receptor proteins. This is the first report of isolation of TSH receptor molecules from nonmammalian vertebrates. PMID- 11006116 TI - Amplification and overexpression of TGIF2, a novel homeobox gene of the TALE superclass, in ovarian cancer cell lines. AB - Homeodomain transcription factors play important roles in directing cellular proliferation and differentiation. A TALE-superclass homeodomain protein, multifunctional repressor of TGFbeta-induced transcription. Here we report identification of TGIF2, a novel TALE-superclass homeodomain protein that shows distinct homology with TGIF, especially in its DNA-binding domain. TGIF2 is expressed ubiquitously in human tissues, with the highest levels being found in heart, kidney, and testis. The TGIF2 product contains a putative nuclear localization signal; translocation of the protein to the nucleus was confirmed by transfection of epitope-tagged cDNA. TGIF2 lies on chromosome 20q11.2-12. Since amplification of 20q is often observed among ovarian cancers, we determined the status of DNA copy-number and expression of TGIF2 in 14 ovarian-cancer cell lines. This gene was over-expressed in all lines that showed amplification by FISH analysis. The results suggested that TGIF2 may play an important role in the development and/or progression of some ovarian tumors through a mechanism of gene amplification. PMID- 11006117 TI - Human CDK10 gene isoforms. AB - The CDK10/PISSLRE gene has been shown to encode two different CDK-like putative kinases. The function(s) of the gene products are unknown, although a role at the G2/M transition has been suggested. We characterised two novel cDNAs. CDK10 mRNA quantity was not found to be correlated with cell proliferation status in HeLa or WI38 cell cultures or in human tissues. Relative levels of the four CDK10 isoforms were studied by RT-PCR, of which three were principally expressed. The two initially cloned isoforms predominated in human tissues, except in brain and muscle. Relative isoform levels did not vary during the cell cycle in culture, except when cells entered into the cell cycle. Finally, the predominant isoforms were shown to have different translation initiation sites and to have different subcellular distribution, due to an alternatively spliced nuclear localisation signal. PMID- 11006118 TI - Norcantharidin-induced post-G(2)/M apoptosis is dependent on wild-type p53 gene. AB - Norcantharidin (NCTD), a synthetic analogue of phosphatase type 2A inhibitors, cantharidin, was shown to have limited effects in treating human and animal tumors. The tumor cell killing mechanisms by norcantharidin, however, remain unclear. In this report, we wished to investigate the mechanisms of norcantharidin-mediated cytotoxicity. Effort was made to investigate whether norcantharidin exerted its cytotoxicity through a p53-dependent or -independent mechanism. RT-2 (wtp53) and U251 (mutant p53) glioblastoma cell lines were exposed to norcantharidin at different dosages. Time-course fluorescent-activated cell sorting (FACS) analysis showed that high doses of norcantharidin arrested the cells at the G(2)/M phase and subsequent post-G(2)/M apoptosis in RT-2 cell line. In comparison, the U251 cell line was found resistant to norcantharidin induced cytotoxicity. Restoring wild-type p53 gene function in the U251 cell line after adenoviral infections induced tumor cell cytotoxicity after exposure to norcantharidin. These results showed that norcantharidin kills tumor cells efficiently corresponding to their endogenous p53 gene status. The results also showed the feasibility of using adenoviral p53 gene therapy to enhance chemosensitivity of tumor cells to norcantharidin. PMID- 11006119 TI - Molecular cloning of the crustacean DD4 cDNA encoding a Ca(2+)-binding protein. AB - A cDNA, named DD4, was identified in the prawn Penaeus japonicus in a search for genes that were expressed during calcification of the crustacean exoskeleton. DD4 transcripts were detected in the epidermal cells underlying the exoskeleton specifically during the postmolt stage, when the calcification takes place. In the DD4 cDNA an open reading frame of 542 amino acids was found. The deduced protein was acidic and proline-rich, and exhibited similarity to the Drosophila Ca(2+)-binding protein calphotin in the amino acid sequence and composition. The DD4 cDNA was expressed in Escherichia coli to characterize Ca(2+)-binding of the encoded protein, and Ca(2+) was found to bind to a central segment of 186 amino acids. The DD4 protein is suggested to play a role in the calcification of the crustacean exoskeleton. PMID- 11006120 TI - Enzymatic modification of heparan sulfate on a biochip promotes its interaction with antithrombin III. AB - A heparan sulfate glycosaminoglycan chain, biotinylated at its reducing-end, was bound to a streptavidin-coated biochip. Surface plasmon resonance spectroscopy showed a low affinity interaction with antithrombin III (ATIII) when it was flowed over a surface containing heparan sulfate. ATIII bound tightly with high affinity when the same surface was enzymatically modified to using 3-O sulfotransferase isoform 1 (3-OST-1) in the presence of 3'-phosphoadenosine 5' phosphosulfate (PAPS). The 3-OST-1 enzyme is involved in heparan sulfate biosynthesis and introduces a critical 3-O-sulfo group into this glycosaminoglycan affording the appropriate pentasaccharide sequence capable of high affinity binding to ATIII. This experiment demonstrates the specific structural modification of a glycosaminoglycan bound to a biochip using a biosynthetic enzyme, suggesting a new approach to rapid screening glycosaminoglycan-protein interactions. PMID- 11006121 TI - A novel form of gonadotropin-releasing hormone in the medaka, Oryzias latipes. AB - The present study has identified three molecular forms of gonadotropin-releasing hormone (GnRH) in the brain of a teleost, the medaka, by isolation of their cDNAs. This species has a novel GnRH, which is here named medaka-type GnRH (mdGnRH), in addition to two characterized forms, chicken-II-type GnRH (cGnRH-II) and salmon-type GnRH (sGnRH). Phylogenetic analysis showed that mdGnRH is a medaka homolog of and seabream-type GnRH (sbGnRH) and mammalian-type GnRH (mGnRH) in other species, and suggested that all vertebrates have three distinct GnRHs. Furthermore, in situ hybridization revealed that the mdGnRH gene is expressed only in neurons clustered within the preoptic area as sbGnRH and mGnRH genes in other species are, while the genes for cGnRH-II and sGnRH are only in the midbrain tegmentum and nucleus olfactoretinalis, respectively. This result suggested that mdGnRH is a hypophysiotropic factor and the other two forms are involved in other physiological events as neuromodulators or neurotransmitters. PMID- 11006122 TI - Biological characterization of uncleavable plasma membrane-anchored human macrophage colony-stimulating factor. AB - The cell-surface form of human macrophage colony-stimulating factor (CSF-1(256), M-CSFalpha) is a plasma membrane-anchored transmembrane protein from which the soluble CSF-1 is released by ectodomain proteolytic cleavage. We have previously generated two forms of cell surface CSF-1 which failed to undergo the cleavage by deleting residues 161-165 or residues 159-165 in the extracellular juxtamembrane region (1). To determine the biologic significance of the ectodomain cleavage, we compared the biosynthesis and biologic activities of uncleavable mutant CSF-1 forms with those of the cleavable wild-type (WT) CSF-1. We found that the uncleavable CSF-1 forms were able to accumulate on cell surface at about threefold higher level than the cleavable WT CSF-1 did. We further demonstrated that the uncleavable plasma membrane-anchored forms of CSF-1 were biologically active in mediating the proliferation of CSF-1-dependent cells as well as the intercellular adhesion between CSF-1 receptor-bearing cells and CSF-1 expressing cells. Furthermore, the adhesive activity of uncleavable CSF-1 forms was about twofold stronger than that of WT CSF-1, which indicated that the ectodomain cleavage system plays an important role in regulating the biologic activities of membrane-anchored CSF-1. PMID- 11006123 TI - An anion channel in guinea pig gallbladder epithelial cells is highly permeable to HCO(-)(3). AB - In guinea pig gallbladder epithelium, a secretion of fluid, secondary to an electrogenic secretion of Cl(-) and HCO(-)(3), is elicited in the presence of a high intracellular concentration of adenosine 3'-5'-cyclic monophosphate (cAMP). The aim of this study was to analyze the effects of secretagogues on the activity of anionic channels in isolated epithelial cells using the patch-clamp technique and measuring the electrical potential difference of the cellular membrane (pd(cm)). In cell-attached configuration, with the microelectrode filled with a solution of N-methylglucamine-Cl, or in inside-out configuration (symmetrical solution), it was possible to demonstrate the presence of an 18-pS Cl(-) channel with linear current/voltage (I/V) relationship and voltage independence; this channel is not activated by cAMP (cell-attached configuration). In inside-out configuration (symmetrical solution), another anionic channel with a conductance of 2.8 pS, voltage independence, and a linear I/V relationship was also identified. This channel was stimulated by cAMP (cell-attached configuration) and by PKA + ATP + cAMP (inside-out configuration). The channel was inhibited by NPPB (10(-5) M), but not by other anionic inhibitors. Measurements of the pd(cm) value suggested that in isolated cells, as in whole tissue, cAMP activates conductance for both Cl(-) and HCO(-)(3). The selectivity of the channel was gluconate < SO(2 )(4) < Cl(-) < Br(-) < I(-) < HCO(-)(3) < SCN(-) and the P(HCO(3))/P(Cl) was 2.6. Some features of the channel resemble those of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel and RT-PCR performed on mRNA from isolated epithelial cells detected the presence of a CFTR homologue mRNA. The results obtained indicate that this channel is responsible for the HCO(-)(3) conductance activated by cAMP. PMID- 11006124 TI - Hypoxia induces the expression of a 43-kDa protein (PROXY-1) in normal and malignant cells. AB - This study was designed to determine the expression of cellular factors that may participate in phenotypic changes that occur under conditions of hypoxia. Using the RT-PCR differential display method, we isolated a cDNA fragment corresponding to a gene whose expression was induced in trophoblast and breast carcinoma cells cultured under 1 or 2% oxygen vs 4% oxygen or higher. This gene encodes a 43-kDa protein initially identified in homocysteine-treated endothelial cells and later shown to be upregulated in various human and mouse cell types (termed RTP, Drg1, Cap43, rit42, Ndr1). Herein we refer to this gene product as PROXY-1, for Protein Regulated by OXYgen-1. Elevated mRNA and protein levels were first observed in cells cultured in 1% oxygen for 8 h. Although PROXY-1 mRNA levels returned to near-control values within 2 h of reexposure to 20% oxygen, protein levels remained high 72 h after reexposure to 20% oxygen. Treatment of cells with hypoxia mimics such as cobalt or iron chelators also increased PROXY-1 expression. Moreover, presence of 30% carbon monoxide in the hypoxic atmosphere abrogated the upregulation of PROXY-1 expression. These findings suggest that hypoxia upregulates PROXY-1 levels through a heme protein-dependent pathway and that assessment of PROXY-1 expression may be of potential use in evaluating tissue hypoxia. PMID- 11006125 TI - Neuroendocrine-specific protein C, a marker of neuronal differentiation, is reduced in brain of patients with Down syndrome and Alzheimer's disease. AB - Neuroendocrine-specific protein C (NSP-C) is found in neural and neuroendocrine cells and associated with the endoplasmic reticulum. Its expression was found to correlate with the degree of neuronal differentiation. As the neuropathological findings in Down syndrome (DS) includes deficits of differentiation, and we detected a downregulated sequence with 100% homology with NSP-C homolog mRNA in temporal cortex of patients with DS as well as Alzheimer's disease (AD) using differential display-polymerase chain reaction (DD-PCR), we decided to examine the protein levels of NSP-C in temporal, frontal cortex and cerebellum of DS and AD. To normalize NSP-C versus neuronal density, we also determined neuron specific enolase (NSE) levels and calculated the ratios. NSP-C was significantly reduced in DS (temporal and frontal cortex) and AD (frontal cortex) compared to controls. The significant decrease of NSP-C in DS was even more pronounced when related to NSE levels. Impaired differentiation in DS brain may well be due to absolutely and relatively decreased NSP-C levels in temporal and frontal cortex. As NSP-C was also reduced in AD frontal cortex, NSP-C deficits in these disorders may be reflecting neurodegenerative changes rather than a primary and specific finding of DS or AD pathogenesis. PMID- 11006126 TI - Identification and characterization of rat AILIM/ICOS, a novel T-cell costimulatory molecule, related to the CD28/CTLA4 family. AB - Activation-inducible lymphocyte immuno-mediatory molecule (AILIM) is an inducible cell surface glycoprotein expressed on thymocytes and activated lymphocytes. Specific monoclonal antibody to rat AILIM induced the cell aggregation of a rat thymoma cell line and ConA-activated splenocytes. In the present study, we identified the primary structure of two species of rat AILIM by expression cloning. We also cloned mouse and human AILIM homologues and the predicted amino acid sequences were identical to those of the inducible costimulator ICOS/CRP-1, which belongs to the CD28/CTLA4 family. Although the human and mouse AILIM/ICOS molecule is localized on T-cells, the major population of AILIM/ICOS-positive cells in rat splenocyte was CD45RA-positive B-cells. The expression level of AILIM/ICOS on T-cells was relatively low; however, its expression was drastically induced by the treatment with PMA plus Ca-ionophore or the engagement of CD3 and these costimulatory molecules. Almost all T-cells exhibited potency as to its expression. Functional analysis of AILIM/ICOS demonstrated that AILIM-mediated costimulation was relatively weak compared to that of human. PMID- 11006127 TI - Enhancement of the activity of l-aspartase from Escherichia coli W by directed evolution. AB - l-Aspartase was modified by directed evolution. After four rounds of error-prone PCR and three rounds of DNA shuffling, an evolved enzyme purified from the final round showed a 28-fold increased k(cat)/K(m) and 4.6-fold decreased K(m). The thermostability and stable pH range were also enhanced. The DNA sequence of the evolved aspartase gene showed seven base changes, resulting in three amino acid changes from the native enzyme: N217K, T233R, V367G. The mechanism of the enhancement of activity was analyzed. PMID- 11006128 TI - Receptor isoform-specific interaction of prostaglandin EP3 receptor with muskelin. AB - By using the yeast two-hybrid system, muskelin was found to bind with the carboxy terminal tail of the prostaglandin EP3 receptor alpha isoform but not with either the beta or gamma isoform. A direct interaction between the carboxy-terminal tail of the alpha isoform and muskelin was confirmed in vitro using recombinant fusion proteins. Analysis by confocal microscopy indicated that the isoform and muskelin were distributed at the plasma membrane in transfected cells. When the isoform was stimulated by agonist, the receptor was internalized in the cells expressing the receptor alone, but this internalization was partially inhibited by the cotransfection with muskelin. Furthermore, muskelin enhanced the Gi activity of the isoform. Thus, muskelin appears to be an isoform-specific anchoring protein for the EP3 receptor. PMID- 11006129 TI - Drosophila von Hippel-Lindau tumor suppressor complex possesses E3 ubiquitin ligase activity. AB - Mutations of the von Hippel-Lindau (VHL) tumor suppressor gene predispose individuals to a variety of human tumors, including renal cell carcinoma, hemangioblastoma of the central nervous system, and pheochromocytoma. Here we report on the identification and characterization of the Drosophila homolog of VHL. The predicted amino acid sequence of Drosophila VHL protein shows 29% identity and 44% similarity to that of human VHL protein. Biochemical studies have shown that Drosophila VHL protein binds to Elongins B and C directly, and via this Elongin BC complex, associates with Cul-2 and Rbx1. Like human VHL, Drosophila VHL complex containing Cul-2, Rbx1, Elongins B and C, exhibits E3 ubiquitin ligase activity. In addition, we provide evidence that hypoxia inducible factor (HIF)-1alpha is the ubiquitination target of both human and Drosophila VHL complexes. PMID- 11006130 TI - Upregulation of the apoptosis-associated protein Grb3-3 in HIV-1-infected human CD4(+) lymphocytes. AB - The mechanism(s) by which HIV-1 infection contributes to depletion of CD4(+) T cell is not well understood. In this report, we investigated whether a recently identified isoform of growth factor receptor bound protein (Grb2), named Grb3-3, a signaling molecule that is associated with the MAP kinase pathway and with apoptosis could be involved. We find that Grb3-3 is markedly up-regulated following HIV-1 infection of CD4(+) peripheral blood mononuclear cells undergoing apoptosis. Although IL-2 deprived CD4(+) cells also undergo apoptosis to a similar extent, Grb3-3 upregulation is not detected under these experimental conditions. Transient overexpression of Grb3-3 in Jurkat T-cells also causes apoptosis. Upon staurosporine stimulation, Grb3-3 predisposes Sup-T1 cell to apoptosis. Finally, analysis of the HIV-1 genes responsible for Grb3-3 expression demonstrates that Tat and Nef can independently induces its expression, suggesting these two earliest viral gene products of HIV-1 may share some common pathway(s) in up-regulating Grb3-3 expression. PMID- 11006131 TI - Anti-angiogenic potential of a cancer chemopreventive flavonoid antioxidant, silymarin: inhibition of key attributes of vascular endothelial cells and angiogenic cytokine secretion by cancer epithelial cells. AB - In recent studies, we have shown that silymarin, a naturally occurring flavonoid antioxidant, exhibits anti-cancer effects against several epithelial cancers. Here, we assessed its potential as an anti-angiogenic agent employing human umbilical vein endothelial cells (HUVEC) and human prostate and breast cancer epithelial cells. When sub-confluent HUVEC were treated for 48 h, adherent cell number decreased by 50 and 90% at 50 and 100 microg/ml doses, respectively. Apoptotic cell death principally accounted for cell loss at >50 microg/ml doses. In biochemical analysis, silymarin treatment of HUVEC for 6 h resulted in a concentration-dependent decrease in the secretion and cellular content of matrix metalloproteinase (MMP)-2/gelatinase A. Silymarin also inhibited HUVEC tube formation (in vitro capillary differentiation) on a reconstituted extracellular matrix, Matrigel. In other studies, 5 to 6 h exposure of DU145 prostate, and MCF 7 and MDA-MB-468 breast cancer cells to silymarin resulted in a dose-dependent decrease in the secreted vascular endothelial growth factor (VEGF) level in conditioned media without any visible change in cell morphology. The inhibitory effect of silymarin on VEGF secretion occurred as early as 1 h. These observations indicate a rapid inhibitory action of silymarin on the secretion of this primary angiogenic cytokine by cancer epithelial cells. Taken together, the results of this study support the hypothesis that silymarin possesses an anti angiogenic potential that may critically contribute to its cancer chemopreventive efficacy. PMID- 11006132 TI - The endogenous oxindoles 5-hydroxyoxindole and isatin are antiproliferative and proapoptotic. AB - Oxindole-core synthetic molecules are currently being developed as anticancer drugs that target protein tyrosine kinases associated with growth factor receptors. Oxindole, 5-Hydroxyoxindole, and 2, 3-dioxindole [isatin] are natural molecules found in mammalian body fluids and tissues and we addressed the question of similar properties of endogenous oxindoles. 5-Hydroxyoxindole and isatin, but not oxindole, inhibited N1E-115, BALB/c3T3, BBC, PC12, and HL60 proliferation at submicromolar concentrations. Acute treatment with 5 hydroxyoxindole and isatin reduced the activity of extracellular signal regulated protein kinases (ERKs) by 35% at 100 microM and ERK1 activity was strongly inhibited by 5-Hydroxyoxindole at 10 microM. Survival of PMA-differentiated HL60 and FGF(2)-differentiated PC12 cells was not affected by 5-Hydroxyoxindole and isatin treatment, suggesting that endogenous oxindoles interact with growth factors signaling. The physiological implications of these data and the potential utility of 5-Hydroxyoxindole and isatin as antitumor agents are discussed. PMID- 11006133 TI - Physical interaction of Delta1, Jagged1, and Jagged2 with Notch1 and Notch3 receptors. AB - The Delta/Serrate/LAG-2 (DSL) domain-containing proteins, Delta1, Jagged1, and Jagged2, are considered to be ligands for Notch receptors. However, the physical interaction between the three DSL proteins and respective Notch receptors remained largely unknown. In this study, we investigated this issue through the targeting of Notch1 and Notch3 in two experimental systems using fusion proteins comprising their extracellular portions. Cell-binding assays showed that soluble forms of Notch1 and Notch3 proteins physically bound to the three DSL proteins on the cell surface. In solid-phase binding assays using immobilized soluble Notch1 and Notch3 proteins, it was revealed that each DSL protein directly bound to the soluble Notch proteins with different affinities. All interactions between the DSL proteins and soluble Notch proteins were dependent on Ca(2+). Taken together, these results suggest that Delta1, Jagged1, and Jagged2 are ligands for Notch1 and Notch3 receptors. PMID- 11006134 TI - Farnesyl transferase inhibitors: current developments and future perspectives. AB - Ras oncogenes play an important role in carcinogenesis and are frequently found in various human tumour types. Cellular activity of Ras oncoprotein, regulated through the enzyme farnesyl transferase, is crucial in the process of ras dependent carcinogenesis, and therefore, specific inhibition of this enzyme is an attractive goal in anticancer treatment. Specific inhibitors of farnesyl transferase have been developed in recent years, many of them showing in vitro and in vivo growth inhibitory or cytostatic activity. Recently, results of the first clinical studies with various farnesyl transferase inhibitors have been presented. In the design of phase I and II studies, either single-agent or combination studies, new endpoints have to be defined in order to properly assess feasibility, antitumour activity and clinical valuability. PMID- 11006135 TI - Antisense therapy in cancer. AB - This review discusses laboratory and clinical studies of antisense oligodeoxynucleotides as potential treatments for haematological malignancies and solid tumours. Mechanisms of action, pharmacokinetics, toxicities and potential clinical applications of these agents are described. PMID- 11006136 TI - A review of angiogenesis and antiangiogenic therapy with thalidomide in multiple myeloma. AB - Angiogenesis is the formation of new blood vessels and occurs physiologically during embryonal growth, wound healing and during the menstrual cycle. It is essential for the proliferation and metastases of most malignant neoplasms. Recent evidence suggests that angiogenesis is increased in multiple myeloma and has prognostic value in the disease. Angiogenic cytokines such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor are expressed by myeloma cells and appear to play a role in the increased angiogenesis seen in myeloma. In addition, VEGF may serve as a paracrine growth factor for myeloma cells. Based on the increased angiogenesis observed in myeloma, thalidomide has been studied as antiangiogenic therapy. Although its mechanism of action in myeloma is still unclear, thalidomide appears to be active in 25-30% of patients with refractory myeloma. Major toxicities include constipation, sedation, skin rash, fatigue, and peripheral neuropathy. Studies are ongoing to determine its role as initial treatment for myeloma. This paper reviews the available data on angiogenesis in myeloma, and summarizes the role of thalidomide therapy in this disease. The pharmacology and toxicity of thalidomide are also discussed. PMID- 11006137 TI - The effect of chemotherapy on the growing skeleton. AB - With the increasing use of high dose (poly)chemotherapy schedules in the treatment of childhood cancer it is particularly important to know the adverse effects of these treatments. Growth is a complex mechanism affected not only by chemotherapy but also by the malignancy itself as well as nutritional status, the use of corticosteroids and (cranial) radiation. In vitro and animal studies are often the most useful in determining the effect of a single chemotherapeutic agent on the growing skeleton. In vitro studies have shown doxorubicin, actinomycin D and cisplatin to have a direct effect on growth plate chondrocytes that in animals results in decreased growth and final height. Clinical studies with multiagent chemotherapy have demonstrated that antimetabolites decrease bone growth and final height. Childhood cancer survivors are at risk of a reduced bone mineral density, mainly due to methotrexate, ifosfamide and corticosteroids. This reduced bone mineral density persists into adult life and may increase bone fracture risk at an older age. PMID- 11006139 TI - Articles to appear in forthcoming issues of cancer treatment reviews PMID- 11006138 TI - Treatment-related leukaemia--a clinical and scientific challenge. AB - The development of a second tumour, including treatment-related leukaemia (TRL), is the most devastating complication of intensive cancer chemotherapy. This is especially relevant in the paediatric population as over 70% of children diagnosed with a malignancy will now live at least 5 years. Most TRLs are myeloid leukaemias and carry an overall poor prognosis when compared with their de novo counterparts. Despite the well known association with specific cytotoxic agents, improved understanding of the pathogenesis and risk factors of TRL is ultimately essential if we are to develop successful strategies for prevention and treatment. Here we review these aspects, together with the clinical and diverse biological features of this complication and the efficacy of current therapy. PMID- 11006140 TI - Metabolic factors affecting the reproductive axis in male sheep. AB - Changes in food intake affect the reproductive axis in both sexes, and the nutritional signals involved and the sites that receive those signals are now beginning to be unravelled. Our studies have focussed on the mature male sheep, a model in which high food intake stimulates GnRH-LH pulse frequency for only 10-20 days but continues to promote testicular growth over several months. Different signals and different target organs seem to be responsible for these short- and long-term responses. Short-term dietary treatments lead to changes in blood concentrations of glucose, fatty acids, insulin and leptin, and concentrations of glucose, insulin, leptin and some amino acids in cerebrospinal fluid. It seems unlikely that amino acids affect GnRH-LH secretion directly in sheep. Intracerebroventricular infusions of insulin specifically increase LH pulse frequency, but intravenous, intra-abomasal or intracerebroventricular infusions of glucose have no effect, despite their effects on cerebrospinal fluid insulin concentrations. The addition of fatty acids to the diet also increases LH pulse frequency, but does not affect the concentrations of insulin or leptin in the cerebrospinal fluid. It appears that acute responses to changes in nutrition involve a range of alternative pathways, possibly including interactions among insulin, leptin and energy substrates. Effects of long-term dietary treatments on testicular size are only partly dependent on the GnRH-LH system (that is, on brain control) and so must also depend on other, as yet unknown, pathways. Concepts of 'metabolic sensing and integration' are being developed from the basis of existing knowledge of the central control of appetite and reproduction. PMID- 11006141 TI - Effect of efferent duct ligation on the function of the blood-testis barrier in rats. AB - The function of the blood-testis barrier has been assessed from the ratio of the Cr-EDTA space in the parenchyma to the measured interstitial volume in the testes of rats at various times after unilateral ligation of the efferent ducts. The barrier remained effective during the phase of fluid accumulation and testicular mass gain, which was linear for at least 24 h, but the testis mass began to decrease between 32 and 40 h after efferent duct ligation, and the Cr-EDTA space at 40 and 48 h after efferent duct ligation exceeded the volume of the interstitial tissue. This finding indicated that, at these times, the barrier to Cr-EDTA, which is normally excluded from the tubules, had broken down and the marker was entering the tubules. Thereafter, the Cr-EDTA space decreased again to be less than the interstitial tissue volume, indicating a restoration of the barrier function, although degeneration of the seminiferous epithelium continued to become more obvious. The present study is the first report of a reversible breakdown of the barrier, but the relevance of the breakdown to the effects on spermatogenesis requires further study. PMID- 11006142 TI - Effectiveness of zona pellucida protein ZPB as an immunocontraceptive antigen. AB - Immunization of female mammals with native zona pellucida (ZP) proteins is known to cause infertility. Since each human ZP protein is now available as a purified recombinant protein, is it possible to compare the immunocontraceptive potential of each ZP protein. A breeding study was conducted in cynomolgus monkeys (Macaca fasicularis) after immunization with recombinant human ZP (rhZP) proteins (ZPA, ZPB, ZPC) separately and in combinations. This study demonstrated that immunization with recombinant human ZPB (rhZPB) protein caused cynomolgus monkeys to become infertile for 9-35 months. A second study was conducted in baboons (Papio cynocephalus), which yielded a similar result. The baboons immunized with rhZPB became infertile for 9 to > 20 months. During the time of maximum antibody titre, some animals experienced disruption of the menstrual cycle, but eventually all of the animals resumed normal menstrual cycles. Control animals and animals immunized with other rhZP proteins all became pregnant before any of the rhZPB treated animals. This is the first study in which a recombinant ZP protein has consistently induced infertility in a primate without permanent disruption of the normal menstrual cycle. PMID- 11006143 TI - Immunohistochemical localization of prostaglandin G/H synthase 1 and 2 in sheep placenta after glucocorticoid-induced and spontaneous labour. AB - Enhanced prostaglandin production and release by the placenta is an essential element in the normal transition to labour in many animal species. In sheep, expression of prostaglandin G/H synthase (PGHS) is the central enzyme regulating this process. In this study immunohistochemistry was used to examine the distribution of cells expressing PGHS-1 and PGHS-2 in ovine placenta in association with spontaneous parturition (n = 6) and glucocorticoid-induced labour (n = 5). Labour was induced in ewes after the intrafetal injection of betamethasone on day 131 of gestation. Animals administered an intrafetal injection of isotonic saline (n = 5) acted as non-labour controls. In placentomes collected from all groups, immunoreactive PGHS-1 was present in the mononuclear trophoblast cells of the fetal placenta. Cells in the maternal mesenchyme and epithelial syncytium were weakly immunopositive for this enzyme. PGHS-1 immunoreactivity was also demonstrated in the endothelial cells of the chorionic vessels. The PGHS-2 isozyme was localized exclusively to the trophoblast epithelial cells. Immunoreactive PGHS-2 was not detectable in the maternal epithelial syncytium or the stroma of the cotyledons. The binucleate cells of the fetal placenta were consistently immunonegative for both PGHS isozymes. These results indicate that the cellular localization of PGHS-1 and PGHS-2 in ovine placenta does not change during the last 15 days of pregnancy. Co-localization of these isozymes indicates that the source of arachidonic acid and the site of prostanoid formation are the same. Quantitation of the percentage area of positive staining for PGHS-1 and PGHS-2 using image analysis software demonstrated a significant increase in PGHS-2 in the fetal trophoblast after glucocorticoid-induced labour and spontaneous parturition. This finding indicates that increased formation of the PGHS-2 isozyme is responsible for the large increase in prostaglandin production by the ovine placenta at term labour. PMID- 11006144 TI - Successful capacitation and homologous fertilization in vitro in Calomys musculinus and Calomys laucha (Rodentia - sigmodontinae). AB - Small South American rodents of the genus Calomys have been used extensively for virology and ecological research. Previous studies have demonstrated that Calomys musculinus and Calomys laucha have a relatively short oestrous cycle and that superovulation and parthenogenetic activation can be induced. The purpose of this study was to determine the requirements for in vitro manipulation of the male gamete and in vitro fertilization. Two culture media and different concentrations of spermatozoa were tested for their ability to support sperm motility, hyperactivation and the acrosome reaction. The ability of capacitated Calomys spermatozoa to penetrate zona-free hamster eggs was also evaluated. In vitro fertilization was assessed by examining attachment and binding to the zona pellucida, second polar body extrusion, pronucleus formation and the fertilizing sperm tail. The results of the study showed that: (i) Tyrode's albumin lactate pyruvate (TALP) medium was more effective than T6 medium for maintaining sperm motility in vitro; (ii) hyperactivation was achieved with TALP but not with T6; (iii) the acrosome reaction was easily distinguished by light microscopy and depends on time and sperm concentration; (iv) capacitated spermatozoa are able to penetrate zona-free hamster eggs; and (v) superovulated oocytes can be fertilized in vitro. This is the first report of capacitation and in vitro fertilization for Calomys sp. These results provide opportunities to use C. musculinus and C. laucha as new laboratory animals for research into reproductive biology. PMID- 11006145 TI - Studies of the oestrous cycle, oestrus and pregnancy in the koala (Phascolarctos cinereus). AB - As an integral part of the development of an artificial insemination programme in the captive koala, female reproductive physiology and behaviour were studied. The oestrous cycle in non-mated and mated koalas was characterized by means of behavioural oestrus, morphology of external genitalia and changes in the peripheral plasma concentrations of oestradiol and progestogen. The mean (+/- SEM) duration of the non-mated oestrous cycle and duration of oestrus in 12 koalas was 32.9 +/- 1.1 (n = 22) and 10.3 +/- 0.9 (n = 24) days, respectively. Although the commencement of oestrous behaviour was associated with increasing or high concentrations of oestradiol, there were no consistent changes in the morphology or appearance of the clitoris, pericloacal region, pouch or mammary teats that could be used to characterize the non-mated cycle. As progestogen concentrations remained at basal values throughout the interoestrous period, non mated cycles were considered non-luteal and presumed anovulatory. After mating of the 12 koalas, six females gave birth with a mean (+/- SEM) gestation of 34.8 +/- 0.3 days, whereas the remaining six non-parturient females returned to oestrus 49.5 +/- 1. 0 days later. After mating, oestrous behaviour ceased and the progestogen profile showed a significant increase in both pregnant and non parturient females, indicating that a luteal phase had been induced by the physical act of mating. Progestogen concentrations throughout the luteal phase of the pregnant females were significantly higher than those of non-parturient females. Parturition was associated with a decreasing concentration of progestogen, which was increased above that of basal concentrations until 7 days post partum. PMID- 11006146 TI - Natural and artificial methods for inducing the luteal phase in the koala (Phascolarctos cinereus). AB - An experiment was conducted in which female koalas were mated for different durations of intromission and ejaculation to confirm that the luteal phase of the oestrous cycle in koalas is induced by the physical act of mating. Results showed that induction of a luteal phase in the koala usually required a complete duration of penile thrusting behaviour from the male. It is proposed that induction of a luteal phase in koalas may involve a copuloceptive reflex, triggered by the thrusting of the male's penis into the female's urogenital sinus. Although interrupted mating in koalas may be used to induce a luteal phase in preparation for an artificial insemination programme, this study showed that there is a 12.5% probability that pregnancy will result from semen prematurely emitted by the teaser male. A dose of 250 iu hCG was administered intramuscularly to eight oestrous females to determine whether it was possible to induce a luteal phase artificially. In contrast to control females, which received sterile saline injections, all females injected with hCG showed a significant increase in progestogen concentration above that of basal values, indicating that a luteal phase had been induced successfully. PMID- 11006147 TI - Potential role of alphav and beta1 integrins as oocyte adhesion molecules during fertilization in pigs. AB - Integrin molecules are cell adhesion molecules that are thought to be involved in sperm-oocyte interaction in rodents and humans. The objective of this study was to evaluate whether integrin molecules were present on the surface of pig oocytes, consistent with involvement in sperm-oocyte interaction in this species. Immunocytochemistry and confocal microscopy were used to evaluate the presence of beta1, and alpha1, alpha2, alpha3, alpha4, alpha5, alpha6 and alphav integrin subunits on the plasma membrane of pig oocytes. The beta1 and alphav integrin subunits were present consistently at the surface of pig oocytes; however, the remaining alpha integrin subunits evaluated were not routinely detected. The antibodies to the beta1 and alphav integrin subunits recognized appropriately sized protein bands on western blots of partially purified oocyte plasma membrane. These two antibodies also recognized oocyte plasma membrane protein isolated from a sperm plasma membrane affinity column. Sperm plasma membrane proteins of 137 and 93 kDa appeared to be the ligands for the beta1 integrin subunit as revealed by a western sandwich blot. Antibody to an extracellular domain of the beta1 integrin subunit reduced pig sperm-oocyte binding (P < 0.05), also indicating an assisting role for a beta1 oocyte integrin subunit in sperm oocyte interaction in pigs. These results are consistent with an alphavbeta1 pig oocyte integrin interacting with a ligand on the sperm plasma membrane during fertilization. PMID- 11006148 TI - Production of tissue inhibitors of metalloproteinases (TIMPs) by pig ovarian cells in vivo and the effect of TIMP-1 on steroidogenesis in vitro. AB - Precisely which ovarian cells produce tissue inhibitors of metalloproteinases (TIMPs) is unclear. Although granulosa cells are reported to produce TIMPs, thecal TIMP production has not been investigated nor has the influence of TIMPs on theca cells. Furthermore, although periovulatory follicles have been examined, little is known about smaller ovarian follicles. Follicles >/= 2 mm in diameter were collected from Large White hybrid gilts on the day before predicted oestrus (n = 3) or after hCG treatment (n = 3) and divided into 1 mm size classes. Small (2 to < 5 mm) follicles were kept intact, whereas follicles >/= 5 mm were separated into follicular fluid, granulosa and theca cell compartments. After homogenization, TIMP-1, -2 and -3 were detected by reverse zymography. Theca cells (50 x 10(3) per well) were cultured with TIMP-1 (10, 100 or 200 ng ml(-1) with or without long-R3 insulin-like growth factor I (IGF-I)) in a serum-free system to investigate the effect on steroidogenesis and the number of cells. Both large and small pig follicles produced TIMPs and TIMP-1, -2 and -3 were detected in follicular fluid, granulosa and theca cell samples. There was a phase x tissue type interaction for the presence of both TIMP-1 and -2 (P < 0.03, P < 0.05, respectively), and TIMPs were detected in more granulosa and theca cell samples after hCG than during the follicular phase. The concentrations were influenced by the type of tissue (TIMP-1, P < 0.005; TIMP-2, P < 0.005, TIMP-3, P > 0.05), and the highest concentrations occurred in the theca tissue. There were tissue type x follicle size interactions for the presence of both TIMP-1 and -2 (P < 0.001). In vitro, TIMP-1 increased thecal steroidogenesis after 144 h (oestradiol, P < 0.05, progesterone, P < 0.001) but reduced the number of viable cells (P < 0.001). In conclusion, TIMP-1, -2 and -3 were present in large and small pig follicles and were produced by both granulosa and theca cells, although concentrations differed with the type of tissue. Production was regulated by factors including follicle size and phase of the oestrous cycle. In addition to controlling tissue remodelling, TIMP-1 may also regulate steroidogenesis. PMID- 11006149 TI - Immediate and delayed effects of heat stress on follicular development and its association with plasma FSH and inhibin concentration in cows. AB - The aim of this study was to characterize the immediate effects of heat stress on plasma FSH and inhibin concentrations, and its involvement in follicular dynamics during a complete oestrous cycle, and to examine a possible delayed effect of heat stress on follicular development. Holstein dairy cows were oestrous synchronized and randomly assigned to either cooled (n = 7) or heat-stressed (n = 6) treatment groups. During a complete oestrous cycle, control cows, which were cooled, maintained normothermia, whereas heat-stressed cows, which were exposed to direct solar radiation, developed hyperthermia. At the end of this oestrous cycle (treated cycle), both groups were cooled and maintained normothermia for the first 10 days of the subsequent oestrous cycle. Throughout this period, follicular development was examined by ultrasonography, and plasma samples were collected. During the second follicular wave of the treated oestrous cycle, a significantly larger cohort of medium sized follicles (6-9 mm) was found in heat stressed cows than in cooled cows (P < 0.05). The enhanced growth of follicles in this wave in heat-stressed cows was associated with a higher plasma FSH increase which lasted 4 more days (days 8-13 of the oestrous cycle; P < 0.05), and coincided with a decrease in the plasma concentration of immunoreactive inhibin (days 5-18 of the oestrous cycle; P < 0.05). During the follicular phase (days 17 20 of the treated cycle), heat-stressed cows showed an increase in the number of large follicles (>/= 10 mm), and the preovulatory plasma FSH surge was significantly higher in heat-stressed cows than in cooled cows (P < 0.01). The effect of heat stress was also observed during the first follicular wave of the subsequent cycle: the postovulatory plasma FSH concentration was higher (P < 0.01), but fewer medium follicles developed, and the first follicular wave decreased at a slower rate in previously heat-stressed cows than in cooled cows (0.40 and 0.71 follicles per day, respectively). This study shows both immediate and delayed effects of heat stress on follicular dynamics, which were associated with high FSH and low inhibin concentrations in plasma. These alterations may have physiological significance that could be associated with low fertility of cattle during the summer and autumn. PMID- 11006150 TI - Loss of myometrial oxytocin receptors during oxytocin-induced and oxytocin augmented labour. AB - Oxytocin is used widely for the induction and augmentation of labour, but there is little information about the dynamics of oxytocin receptors in human myometrium during parturition, and the possible effect of oxytocin infusion. This information is important because G protein-coupled receptors, such as the oxytocin receptor, undergo desensitization after prolonged or repeated stimulation. The concentration of myometrial oxytocin receptors and the steady state of its mRNA were measured in patients undergoing Caesarean sections before or during spontaneous or induced labour. The concentration of receptors before labour was 477 (175-641) fmol mg(-1) protein (median, quartile range), and decreased to 140 (72-206; P < 0.05) and 118 (69-75; P < 0.01) fmol mg(-1) protein during prolonged oxytocin-augmented and oxytocin-induced labour, respectively. The corresponding oxytocin receptor mRNA concentrations decreased by 60- and 300 fold, respectively. The decrease in receptor binding and mRNA in women receiving oxytocin infusion indicates that homologous receptor desensitization occurs in vivo. PMID- 11006151 TI - A staging scheme for assessing development in vitro of organogenesis stage embryos of the stripe-faced dunnart, Sminthopsis macroura (Marsupialia: dasyuridae). AB - The inaccessibility of mammalian organogenesis stage embryos has precluded their widespread use in embryological and teratological studies. As organogenesis occurs during the last 1.5 days of the 10. 7 days of gestation in the stripe faced dunnart (Sminthopsis macroura), the aim of the present study was to investigate whether day 9 and day 10 embryos and fetuses could be grown to term in vitro. High glucose Dulbecco's modified Eagle's medium with 10% fetal calf serum (FCS) supported embryonic growth for various periods of time, some to within 5 h of the predicted time of parturition. A roller culture system maintained at 35 degrees C was used to incubate organogenesis stage embryos (n = 43). Nine unincubated (control) embryos were either fixed for microscopic analysis or frozen for microprotein determination. The results of the present study indicate that with some optimization of the culture conditions (increasing oxygen in the gas phase in the culture tubes, replacing FCS with rat serum), it might be possible for organogenesis stage S. macroura embryos to be grown to term. A scoring scheme for assessing morphological development was devised for use as a standard in staging organogenesis stage embryos. This scheme reflects the highly compressed schedule of developmental events that occurs mainly during day 9 of gestation in S. macroura embryos. In comparison, during embryogenesis in Didelphis virginiana these developmental events occur from day 8 to day 10.5 of gestation, and birth occurs on day 13. PMID- 11006152 TI - Insulin-like growth factor I receptor mRNA and protein expression in pig corpora lutea. AB - Insulin-like growth factor I (IGF-I) is believed to play a luteotrophic role in the pig corpus luteum during the oestrous cycle. Since the actions of IGF-I in target tissues are mediated by the type I IGF receptor, the concentrations of IGF I receptor mRNA and protein were examined in pig corpora lutea at different stages of the oestrous cycle. Corpora lutea were collected from normally cyclic gilts on days 4, 7, 10, 13, 15 and 16 of the oestrous cycle (n = 4 animals per day). Corpora lutea on days 7, 10 and 13 were dissociated with collagenase, and large and small luteal cell sub-populations were separated by elutriation. Northern and slot blots were used to examine mRNA, and western blots were used to measure the concentrations of IGF-I receptor protein in the pig corpus luteum. On northern blots, luteal IGF-I receptor mRNA was present as a single 11 kb transcript. The slot blots showed that the steady state expression of IGF-I receptor mRNA increased significantly (P < 0.05) from its lowest value on day 4, to reach a maximum on days 13-16. IGF-I receptor mRNA was also expressed to a greater extent in large compared with small luteal cells (P < 0.05). On western blots, IGF-I receptor appeared as a 95 kDa protein band (beta-subunit) and IGF-I receptor protein concentrations were significantly higher (P < 0.05) on days 4-10 than on days 13-16. Finally, large luteal cells appeared to contain more IGF-I receptor protein than the small luteal cells. In conclusion, since IGF-I receptor was detected in the pig corpus luteum, it is a likely target tissue for IGF-I, especially during the early luteal phase. Furthermore, IGF-I receptor was localized primarily on large luteal cells, thus it is hypothesized that IGF-I may play a paracrine role in the pig corpus luteum. PMID- 11006153 TI - Expression of intercellular adhesion molecule 1 (ICAM-1) on the human oviductal epithelium and mediation of lymphoid cell adherence. AB - The epithelium of the human oviduct expresses the major histocompatibility complex (MHC) class II and shows endocytic properties towards luminal antigens. Therefore, the epithelial cells might behave as antigen-presenting cells, inducing a local immune response. The activation of antigen-specific T cells not only requires presentation of the peptide antigen by MHC class II, but also the presence of co-stimulatory molecules in the antigen-presenting cells. Therefore, the expression of the intercellular adhesion molecule 1 (ICAM-1) was examined in the epithelium of the human oviduct. Most oviducts showed epithelial ICAM-1 expression, as assessed by immunocytochemistry, western blot analysis and RT-PCR assay, and the expression was restricted to the luminal border of ciliated and secretory cells. Interferon gamma, interleukin 1 and lipopolysaccharide treatments increased the percentage of ICAM-1-positive cells in primary cultures, indicating that the expression of ICAM-1 in the oviduct might be upregulated in vivo by inflammatory cytokines or bacterial infections. Binding assays between allogenic phytohaemagglutinin-activated lymphocytes and epithelial monolayers expressing ICAM-1 demonstrated that this molecule stimulated lymphocyte adherence. The presence of ICAM-1, in addition to MHC class II, supports the putative role of the oviductal epithelium in antigen presentation. The exclusive apical distribution of ICAM-1 indicates that T-cell activation would occur in a polarized manner. Binding of lymphoid cells to the surface of the oviductal epithelium may help to retain these immune cells that are required for the clearance of pathogens. PMID- 11006154 TI - Expression of matrix metalloproteinases 2 and 9 in the mouse uterus during implantation and oil-induced decidualization. AB - During implantation, matrix metalloproteinases are believed to play roles in the tissue remodelling that accompanies decidualization in the endometrium and in embryo invasion. The objective of this study was to characterize further the expression of matrix metalloproteinases 2 and 9 in the mouse uterus during early pregnancy and oil-induced decidualization. mRNA encoding matrix metalloproteinase 2 was detected in pregnant uteri and uteri undergoing oil-induced decidualization by northern blot analyses. The steady-state concentrations of mRNA encoding matrix metalloproteinase 2 did not change significantly in implantation compared with inter-implantation areas on days 5-8 of pregnancy but were significantly lower in stimulated compared with non-stimulated uterine horns during artificially induced decidualization. mRNA encoding matrix metalloproteinase 9 was also detected in uteri undergoing oil-induced decidualization but not in pregnant uteri. Its concentration was significantly greater in uterine horns undergoing oil-induced decidualization compared with control horns. Immunoreactive matrix metalloproteinases 2 and 9 were detected in the uterus during early pregnancy and oil-induced decidualization by immunohistochemistry, localized to the endometrial stroma, but the staining progressively became weaker and was absent in areas that had undergone decidualization. By day 8 of pregnancy and 72 h after the induction of decidualization, matrix metalloproteinase 2 and 9 proteins remained mainly in the region of non-decidualized stromal cells adjacent to the myometrium. In implantation segments, they were also localized to the region of the trophoblast giant cells. The second objective of the present study was to determine whether endometrial stromal cells isolated from uteri sensitized for decidualization express matrix metalloproteinases 2 and 9. Northern blot analyses and gelatin zymography showed that these cultured cells expressed matrix metalloproteinase 2 and 9, and that transforming growth factor beta1 significantly increased matrix metalloproteinase 9 expression. The results of the present study further characterize matrix metalloproteinases 2 and 9 expression in the uterus during implantation and artificially induced decidualization. PMID- 11006155 TI - Effects of angiotensin II on the acrosome reaction in equine spermatozoa. AB - Angiotensin II is a hormone with a wide array of physiological effects that exerts its effect via interaction with two major subtypes of receptor. The results of this study show that angiotensin II (from 1 to 100 nmol l(-1)) initiates acrosomal exocytosis in equine spermatozoa that have undergone capacitation in vitro in a TALP-TEST (Tyrode's albumin lactate pyruvate; 188.7 mmol TES l(-1), 84.8 mmol Tris l(-1)) buffer with cAMP. The acrosome reaction and sperm viability were assessed with fluorescein isothiocyanate-Pisum sativum agglutinin (FITC-PSA) and Hoechst 33258, respectively. The initiation of the acrosome reaction by angiotensin II was strongly inhibited by losartan, a specific angiotensin II type 1 receptor antagonist. Although angiotensin II as well as progesterone both initiated the acrosome reaction in equine spermatozoa, there was no synergistic effect when both agonists were added simultaneously. Initiation of acrosomal exocytosis by angiotensin II was accompanied by a rapid and transient calcium influx that was assessed in capacitated spermatozoa loaded with Fura-2AM. In addition, the angiotensin II-mediated calcium influx was inhibited when spermatozoa were preincubated with losartan. Western blotting with an antibody against angiotensin II type 1 receptor detected a major sperm protein of 60 kDa. Indirect immunofluorescence of non-capacitated spermatozoa with the angiotensin II type 1 receptor antibody revealed labelling in the midpiece and tail. In capacitated spermatozoa, the angiotensin II type 1 receptor was localized mainly over the anterior region of the sperm head, the equatorial segment and occasionally on the postacrosomal region in addition to the sperm tail. In conclusion, this study demonstrated the ability of angiotensin II to stimulate the acrosome reaction in capacitated equine spermatozoa. This effect is mediated via the angiotensin II type 1 receptor and is accompanied by an increase in intracellular calcium. PMID- 11006156 TI - Reproductive characteristics of the african pygmy hedgehog, atelerix albiventris. AB - To obtain further perspective on reproduction and particularly gamete function among so-called primitive mammals presently grouped in the Order Insectivora, we have examined the African hedgehog, Atelerix albiventris, in light of unusual features reported in shrews and moles. Atelerix proves to share many but not all of the characteristics seen in these other insectivores. The penis of Atelerix has a 'snail-like' form, but lacks the surface spines common in insectivores and a number of other mammals. Hedgehog spermatozoa display an eccentric insertion of the tail on the sperm head, and they manifest the barbs on the perforatorium that, in shrews, probably effect the initial binding of the sperm head to the zona pellucida. As a possible correlate, the structural matrix of the hedgehog acrosome comprises only two main components, as judged by immunoblotting, rather than the complex of peptides seen in the matrix of some higher mammals. The Fallopian tube of Atelerix is relatively simple; it displays only minor differences in width and in the arborized epithelium between the isthmus and ampulla, and shows no evidence of the unusual sperm crypts that characterize the isthmus or ampulla, depending on the species, in shrews and moles. In common with other insectivores, Atelerix appears to be an induced ovulator, as judged by the ovulation of some 6-8 eggs by about 23 h after injection of hCG. The dense cumulus oophorus appeared to have little matrix, in keeping with the modest dimensions of the tubal ampulla and, while it was not quite as discrete as that of soricids, it did show the same insensitivity to 0.5% (w/v) ovine or bovine hyaluronidase. PMID- 11006157 TI - Control of ovarian follicular growth and maturation by the corpus luteum and the placenta during pregnancy in sheep. AB - Ovarian follicular growth and maturation and its control throughout pregnancy have not been described fully in sheep. Experiment 1 characterized the size and maturation (steroid production in vitro and aromatase activity) of ovarian follicles obtained at days 20, 50, 80 and 110 of pregnancy compared with those obtained at day 12 of the oestrous cycle. There was no difference in the number of small follicles (< 3 mm in diameter) between cyclic and pregnant ewes, regardless of the stage of pregnancy. There was a marked reduction (P < 0.01) in the number of medium follicles (3-5 mm) starting at day 80 of pregnancy. Large follicles (> 5 mm) were not detected at day 110 of pregnancy. In vitro testosterone output by follicles was constant throughout pregnancy. Oestradiol output remained steady until day 80, but decreased markedly at day 110 of pregnancy. This decrease was associated with a reduction in aromatase activity in follicles obtained at this stage. Experiment 2 examined the effect of administration of high concentrations of progesterone between day 100 and day 120 after mating on resumption of follicular growth in ewes that underwent Caesarean section at day 99 of pregnancy. In ewes that underwent Caesarean section, progesterone supplementation was successful in mimicking the profile found in pregnant ewes, but did not prevent re-initiation of follicular growth, as demonstrated by the presence of large follicles (> 5 mm) at day 120 after mating. Experiment 3 examined the effects of PGF(2alpha)-induced regression of the corpus luteum of day 100 of pregnancy on resumption of follicular growth. High concentrations of PGF(2alpha) (0.28 mg kg(-1) body weight) administrated at day 100 of pregnancy were required to initiate regression of the corpus luteum. At day 120 after mating, the mean (+/- SEM) diameter of the largest follicle in PGF(2alpha)-treated ewes (3.40 +/- 0.47 mm) was significantly greater (P < 0.05) than that in control pregnant ewes (2.52 +/- 0.34 mm). Experiment 4 examined the effect of removal of the fetus and of the corpus luteum at day 100 of pregnancy on resumption of ovulation. Removal of the corpus luteum by PGF(2alpha) treatment at the time of removal of the fetus resulted in earlier occurrence of short luteal phases (27.8 versus 40.6 days, PGF(2alpha)-treated versus non-treated) but did not alter the timing of the first normal luteal phases (41 days). In conclusion, the results from these experiments indicate that placental compounds play a major role in inhibiting follicular growth and maturation during late pregnancy in sheep. PMID- 11006158 TI - Gonadotrophin secretion in prepubertal bull calves born in spring and autumn. AB - The reproductive development of bull calves born in spring and autumn was compared. Mean serum LH concentrations in calves born in spring increased from week 4 to week 18 after birth and decreased by week 24. In bull calves born in autumn, mean LH concentrations increased from week 4 to week 8 after birth and remained steady until week 44. LH pulse amplitude was lower in bull calves born in autumn than in calves born in spring until week 24 of age (P < 0.05). There was a negative correlation between LH pulse frequency at week 12 after birth and age at puberty in bull calves, irrespective of season of birth, and LH pulse frequency at week 18 also tended to correlate negatively with age at puberty. Mean serum FSH concentrations, age at puberty, bodyweight, scrotal circumference, testes, prostate and vesicular gland dimensions, and ultrasonographic grey scale (pixel units) were not significantly different between bull calves born in autumn and spring. However, age and body-weight at puberty were more variable for bull calves born in autumn (P < 0.05). In a second study, bull calves born in spring received either a melatonin or sham implant immediately after birth and at weeks 6 and 11 after birth. Implants were removed at week 20. Mean LH concentrations, LH pulse frequency and amplitude, mean FSH concentrations and age at puberty did not differ between the two groups. No significant differences between groups in the growth and pixel units of the reproductive tract were observed by ultrasonography. In conclusion, although there were differences in the pattern of LH secretion in the prepubertal period between bull calves born in autumn and spring, the postnatal changes in gonadotrophin secretion were not disrupted by melatonin treatment in bull calves born in spring. Reproductive tract development did not differ between calves born in spring and autumn but age at puberty was more variable in bull calves born in autumn. LH pulse frequency during the early prepubertal period may be a vital factor in determining the age of bull calves at puberty. PMID- 11006159 TI - Effects of recent sexual experience and melatonin treatment of rams on plasma testosterone concentration, sexual behaviour and ability to induce ovulation in seasonally anoestrous ewes. AB - The aim of this study was to determine whether advancing the seasonal changes associated with rams by treatment with exogenous melatonin and allowing the rams previous sexual experience would increase the proportion of anoestrous ewes ovulating in early July. North Country Mule ewes (n = 225) were grouped by live body weight and body condition score and allocated randomly to the following treatments: (i) isolated from rams (control; n = 25); (ii) introduced to rams (treatment 2); (iii) introduced to rams that had mated with ewes during the previous 2 days (treatment 3); (iv) introduced to rams implanted with melatonin (treatment 4); and (v) introduced to rams that were implanted with melatonin and had mated with ewes during the previous 2 days (treatment 5). Treatments 2-5 were replicated (2 x 25 ewes) and two rams were introduced to each replicate group. Introductions began on 4 July and were completed by 11 July. The rams were withdrawn from the ewes after 8 days. Melatonin was administered as a subcutaneous implant (Regulin((R))) on 22 May and again on 20 June. Blood samples were taken from all rams to determine plasma melatonin and testosterone concentrations (19 samples in 6 h). The behaviour of the sheep was videotaped continuously during the first 3 h after the ram was introduced. Ovulation was detected by an increase in plasma progesterone concentrations from < 0.5 ng ml( 1) to > 0.5 ng ml(-1). Mean +/- SE plasma melatonin concentrations were 649.7 +/- 281.4 and 18.3 +/- 2.4 pg ml(-1) in rams with and without melatonin implants, respectively (P < 0.001). Melatonin implants also increased plasma testosterone concentrations from 4.30 +/- 1.88 to 10.10 +/- 1.10 ng ml(-1) (P < 0.01), the libido of the rams and the proportion of ewes that ovulated in response to the rams (43 and 56% (treatments 4 and 5) versus 24% (treatments 2 and 3)). In conclusion, implanting rams with melatonin before introducing them to seasonally anoestrous ewes increases the proportion of ewes that ovulate in response to introduction of a ram, but previous sexual experience of rams appears to have little or no effect. PMID- 11006160 TI - Effects of bromocriptine administration during the follicular phase of the oestrous cycle on prolactin and gonadotrophin secretion and follicular dynamics in merino monovular ewes. AB - Two experiments using Spanish Merino ewes were conducted to investigate whether the secretion of prolactin during the follicular phase of the sheep oestrous cycle was involved in the patterns of growth and regression of follicle populations. In both experiments, oestrus was synchronized with two cloprostenol injections which were administered 10 days apart. Concurrent with the second injection (time 0), ewes (n = 6 per group) received one of the following treatments every 12 h from time 0 to 72 h: group 1: vehicle injection (control); group 2: 0.6 mg bromocriptine (0.03 mg per kg per day); and group 3: 1.2 mg bromocriptine (0.06 mg per kg per day). In Expt 1, blood samples were collected every 3 h from 0 to 72 h, and also every 20 min from 38 to 54 h to measure prolactin, LH and FSH concentrations. In Expt 2, transrectal ultrasonography was carried out every 12 h from time 0 until oestrus, and blood samples were collected every 4 h to measure prolactin, LH and FSH concentrations. Ovulation rates were determined by laparoscopy on day 4 after oestrus. Bromocriptine markedly decreased prolactin secretion, but did not affect FSH concentrations, the mean time of the LH preovulatory surge or LH concentrations in the preovulatory surge. Both doses of bromocriptine caused a similar decrease in LH pulse frequency before the preovulatory surge. The highest bromocriptine dose led to a reduction (P < 0.01) in the number of 2-3 mm follicles detected in the ovaries at each time point. However, bromocriptine did not modify the total number or the number of newly detected 4-5 mm follicles at each time point, the number of follicles > 5 mm or the ovulation rate. In conclusion, the effects of bromocriptine on gonadotrophin and prolactin secretion and on the follicular dynamics during the follicular phase of the sheep oestrous cycle indicate that prolactin may influence the viability of gonadotrophin-responsive follicles shortly after luteolysis. PMID- 11006161 TI - Regulation of oxytocin receptor gene expression in sheep: tissue specificity, multiple transcripts and mRNA editing. AB - The increase in uterine oxytocin receptor concentrations over the late luteal phase of the oestrous cycle in sheep is thought to play an important role in the regulation of the duration of the cycle by facilitating the effect of oxytocin on uterine prostaglandin release. Experiments indicated that oxytocin receptor mRNA expression in the endometrium was high at oestrus compared with at days 2, 7 and 12 of the oestrous cycle. The amount of oxytocin receptor mRNA expression in the pituitary gland did not show any significant differences during the oestrous cycle. Oxytocin receptor cDNA was obtained and characterized from ovine uterine endometrium on day 15 of the oestrous cycle, using RT-PCR techniques, to study the mechanisms underlying the resolution of oxytocin receptor expression. The cDNA sequence for the oxytocin receptor gene in sheep was found to be similar to that described previously, except for a difference of seven nucleotides. These nucleotide differences resulted in changes in four of the deduced amino acids in the oxytocin receptor sequence. The heterogeneity of the different sized oxytocin receptor transcripts in sheep is due, at least in part, to the alternative use of polyadenylation sites. Northern hybridization confirmed that the oxytocin receptor gene is expressed in ovine corpus luteum. The investigations on oxytocin receptor gene expression indicate that the patten of oxytocin receptor gene expression in sheep is not only tissue-specific, but also highly function related. Evidence was obtained of mRNA editing in both the coding and the 3' untranslated (3'UTR) regions of oxytocin receptor gene transcripts in ovine endometrium; this was the first demonstration of this phenomenon for oxytocin receptor mRNA. The present results indicate that the observed differences in oxytocin receptor mRNA sequences for the different oxytocin receptor populations in endometrium are due to mRNA editing. mRNA editing of oxytocin receptor transcripts may be reflected in changes in the amino acid composition of the carboxyl terminus of the receptor, which would explain the differences in the observed responses to an oxytocin challenge. PMID- 11006162 TI - Is the action of inhibin mediated via a unique receptor? AB - The receptor system and the molecular mechanisms by which inhibin acts on its target cells are poorly understood, in contrast to the situation for the structurally related molecule, activin. On the basis of evidence that the biological action of inhibin in a number of systems resembles that of an activin antagonist, it has been contended that inhibin operates by competition for the activin receptor rather than through a specific inhibin receptor. However, mounting evidence indicates that inhibin also interacts with high affinity and specificity with membrane-binding proteins that are likely to be the putative inhibin receptor. PMID- 11006163 TI - Function of steroidogenic factor 1 during development and differentiation of the reproductive system. AB - Intact reproductive development depends on steroids and other endocrine signals. Although recent discoveries have elucidated important steps of sexual differentiation, the intricate mechanisms that regulate the development of steroid-producing tissues remain elusive. In adults, complex feedback mechanisms determine the hypothalamic and pituitary regulation of steroid hormone biosynthesis. Steroidogenic factor 1 (SF-1), an orphan member of the nuclear receptor superfamily of proteins, plays a critical role in development and differentiation of the endocrine and reproductive systems. This review provides an overview of the function of SF-1, its mechanism of action, and a perspective on the interaction of SF-1 with other determinants of sexual differentiation. Whereas SF-1 is essential for the expression of numerous steroidogenic enzymes, its presence may not correlate with steroidogenic function. Moreover, diverse co regulators modulate the influence of SF-1 on gene transcription. The significance of these interactions is discussed in the context of reproductive development and function. Other orphan or ligand-dependent nuclear proteins may share similar mechanisms. PMID- 11006165 TI - Role of mother-young interactions in the survival of offspring in domestic mammals. AB - The defining characteristic of mammals is that females nurse and care for their young; without this, the neonate has no chance to survive. Studies on wild and domestic species show that the neonatal period is the most critical step in the lifetime of a mammal. This review compares three well-studied species (the rabbit, pig and sheep) that differ in their parental strategies and in the problems that neonates have to overcome. As a general trend, mother-young interactions vary according to the maturity of the newborn, and the size of the litter. Neonatal survival relies to a great extent on an environment that is ecologically appropriate for the developmental stage of the neonate, and on optimum interactions with the mother. Adaptive maternal care supposes that the mother provides the basic needs of the neonate: warmth (in pigs and rabbits) or shelter, food, water and immunological protection (via colostrum) and, in some instances, protection from predators and other conspecifics. A major risk facing all neonates, other than the birth process itself, is inadequate colostrum intake owing to delayed suckling or competition with siblings, which leads to starvation, hypothermia or even crushing, as has been observed in pigs. PMID- 11006164 TI - Roles of KIT and KIT LIGAND in ovarian function. AB - Evidence from mouse mutants indicates that the Kit gene encoding KIT, a receptor present on the oocyte and theca cells, and the Mgf gene encoding KIT LIGAND, the ligand of KIT, are important regulators of oogenesis and folliculogenesis. Recently, in vitro cultures of fetal gonads, of follicles and of oocytes have identified specific targets for the KIT-KIT LIGAND interaction. In fetal gonads, an anti-apoptotic effect of KIT-KIT LIGAND interactions on primordial germ cells, oogonia and oocytes has been demonstrated. In postnatal ovaries, the initiation of follicular growth from the primordial pool and progression beyond the primary follicle stage appear to involve KIT-KIT LIGAND interactions. During early folliculogenesis, KIT together with KIT LIGAND controls oocyte growth and theca cell differentiation, and protects preantral follicles from apoptosis. Formation of an antral cavity requires a functional KIT-KIT LIGAND system. In large antral follicles, the KIT-KIT LIGAND interaction modulates the ability of the oocyte to undergo cytoplasmic maturation and helps to maximize thecal androgen output. Hence, many steps of oogenesis and folliculogenesis appear to be, at least in part, controlled by paracrine interactions between these two proteins. PMID- 11006166 TI - Control of the immunological environment of the uterus. AB - The uterine immune axis holds the key to solving major problems in female reproductive health, including infertility, many pathologies of pregnancy, and sexually transmitted disease. The molecular determinants of tolerance and immunity in the reproductive tract are now being identified, and the governing principles are similar to those in other mucosal tissues. Cytokines are implicated as pivotal regulators at important 'decision-making' points in each phase of the induction and elicitation of a response. Indeed, the flexibility to deal appropriately with antigens as disparate as infectious micro-organisms, spermatozoa and the conceptus is likely to be attributable to the sophistication of the cytokine network in driving immune deviation. A better understanding of the factors controlling the development of immune activity in the uterus, particularly the significance of the inductive cytokine environment in determining the destiny of T-lymphocyte responses, will assist the rational design of new therapeutic strategies to treat immune-based reproductive disorders. PMID- 11006167 TI - Growth hormone: a reproductive endocrine-paracrine regulator? AB - Growth hormone (GH) is not classically considered as a reproductive hormone, although a vast literature indicates that it has roles in reproductive function. It is required for sexual differentiation and pubertal maturation and it participates in gonadal steroidogenesis, gametogenesis and ovulation. GH is also required for fetal nutrition and growth during pregnancy and for mammary development and lactation. Although some of these roles reflect the action of GH on the secretion and action of LH and FSH (Chandrashekar and Bartke, 1998), they also reflect direct actions of GH and indirect actions mediated through the local production of insulin-like growth factor I. Moreover, as GH is produced in gonadal and mammary tissues, these actions may reflect local autocrine or paracrine actions of extrapituitary GH, as well as the endocrine actions of pituitary GH. The roles of GH in reproductive function are considered in this review. PMID- 11006168 TI - Advances in spermatogonial stem cell transplantation. AB - Spermatogonial stem cell transplantation was first reported by Ralph Brinster's laboratory in 1994. It has proven to be a technological breakthrough in the study of both stem cells and Sertoli cell-germ cell interactions. This technique can be used to transfer testicular stem cells successfully from one animal to another of the same species (referred to as syngeneic transplants) and sometimes to an animal of a different species (xenogeneic transplants). This transfer technique, combined with developments in cryopreservation, long-term culture, and the enrichment of stem cell populations makes more significant breakthroughs likely in the near future. Ultimately, the application of spermatogonial stem cell transfer will allow transplantation of cultured stem cells manipulated genetically in vitro to give rise to functional male gametes with an altered genotype. This achievement will have applications in basic science, human medicine, and domestic and wild animal reproduction. Although progress toward this goal has been swift, potentially significant barriers, such as the stable incorporation of genetic material into stem cells and immunological responses to the introduced germ cells, remain to be overcome. This article is a review of the scientific advances made since the initial report of successful transplantation in 1994. PMID- 11006169 TI - Role of progesterone in peripheral nerve repair. AB - Progesterone is synthesized in the peripheral nervous system in glial cells. The functions of progesterone are indicated by the findings that it stimulates neurite outgrowth from dorsal root ganglia sensory neurones in explant cultures, accelerates the maturation of the regenerating axons in cryolesioned sciatic nerve, and enhances the remyelination of regenerated nerve fibres. The formation of myelin sheaths around axons is a sexually dimorphic process, as the sheaths are thicker in female than in male regenerating nerves. The progesterone-induced myelination is probably mediated by progesterone receptors, as it is impaired by mifepristone (RU486), a progesterone antagonist. The stimulation of neurite growth in the peripheral nervous system may be mediated by a progesterone metabolite, 5alpha-tetrahydroprogesterone, through GABA(A) receptors. PMID- 11006170 TI - [Recommendations for the prevention of respiratory syncytial virus infections. Standards Committee of the Spanish Society of Neonatology. Board of Directors of the Spanish Society of Neonatology]. AB - Premature babies < = 35 weeks gestation, with or without chronic lung disease (CLD), should be considered high risk population for RSV infection and rehospitalization. RSV monoclonal antibodies (palivizumab) have been found useful in decreasing rates of RSV hospitalization in this patients. Guidelines for their administration include: 1. Recommend their use in premature born between 29-32 weeks gestation without CLD and less than 6 months at entry of RSV station. 2. Strongly recommend their use in premature babies < = 28 weeks gestation or affected with CLD in treatment during last 6 months. These patients should be prophylaxed for two RSV seasons. 3. Prophylaxis among premature babies between 32 35 weeks gestation is not recommended on routine bases. Each case has to be individually analyzed considering risk factors. PMID- 11006171 TI - [Recommendations for basic, advanced and neonatal cardiopulmonary resuscitation. Post-resuscitation stabilization and transport. Spanish Group for Pediatric and Neonatal Cardiopulmonary Resuscitation]. PMID- 11006172 TI - [Recommendations for basic, advanced and neonatal pediatric cardiopulmonary resuscitation. Ethics and cardiopulmonary resuscitation. Spanish Group for Pediatric and Neonatal Cardiopulmonary Resuscitation]. PMID- 11006173 TI - Publication bias in reproductive research. AB - Publication bias is defined as any tendency on the part of investigators or editors to fail to publish study results on the basis of the direction or strength of the findings. This may lead to overestimation of treatment effects in published work. Inappropriate decisions about patient management may result. We investigated what proportion of abstracts at the European Society of Human Reproduction and Embryology (ESHRE) annual meeting eventually reached full publication, what was the time to publication, and which factors might have affected publication. Among the 2691 abstracts of six ESHRE annual meetings, 151 (5.6%) reporting randomized controlled trials (RCT) were identified. Comprehensive searches of electronic databases and handsearching of the two major journals in the field yielded 79 full publications pertaining to these abstracts. Kaplan-Meier analysis estimated 56% of RCT abstracts to be eventually published in full, the median time to publication being 32.5 months. Positive outcome (i.e. significant results) did not affect the publication rate, and neither did sample size, the subject category, or the native language (English/non-English) of the country of origin. Oral presentations resulted in eventual full publication significantly more frequently (69%) than posters (42%). It is concluded that a considerable publication deficit, but not a publication bias, exists for RCT in reproductive research. PMID- 11006174 TI - Clinical andrology: from evidence-base to ethics. The 'E' quintet in clinical andrology. AB - The management of the infertile man should be founded on consensus-based medicine, i.e. the consensual opinion of experts considering evidence-based as well as empirical or experience-based medicine, the effective cumulative rate of successful deliveries, ethical and economic considerations. The apparent contradictions between conclusions from experience-based medicine and evidence based medicine regarding the efficacy of varicocele treatment and tamoxifen treatment can be explained by scientific reasons. It is argued that the suggestion not to implement these treatments is ill founded because of flawed meta-analyses. The effective cumulative rate of successful deliveries and time to pregnancy as observed in cohort studies should be considered the ultimate touchstone of treatment efficacy. Based on the data of effective cumulative delivery rate, cost per successful delivery, and the known prevalence of aetiological diagnoses in infertile men, it is possible to estimate the number of deliveries that can be attained thanks to an investment of, e.g. 1 million Euro. This number is approximately 70-80 if IVF (including intracytoplasmic sperm injection) is chosen as first line treatment, and four times higher if conventional treatment (including intrauterine insemination) is applied. It is concluded that the well thought out approach recommended by the World Health Organization should generally be implemented for the management of couples in whom infertility is (mainly) due to a male factor. PMID- 11006175 TI - Hydrosalpinx and ART. Salpingectomy prior to IVF can be recommended to a well defined subgroup of patients. AB - The result of the Scandinavian multicentre study on salpingectomy prior to IVF has promoted a discussion on whether there is a risk of unnecessary salpingectomies being performed. We agree that physicians have to discriminate carefully between a hydrosalpinx that should be removed and one that is suitable for surgical repair. Tubal endoscopy is the most advanced tool for that purpose, while transvaginal ultrasound is not appropriate in selecting patients to undergo either salpingectomy or salpingostomy. The Scandinavian study showed that patients with hydrosalpinges which are large enough to be visible on ultrasound before ovarian stimulation, benefit from salpingectomy in terms of improved fertility outcome after IVF. The result of the study does not interfere with the management of distal tubal infertility in general, since it was concluded that not every woman with a large hydrosalpinx should undergo salpingectomy. Hydrosalpinges with preserved mucosa may be better treated with reconstructive surgery as primary treatment instead of salpingectomy and IVF. The latter option may, however, be the secondary treatment after failed conception and re-occlusion of the tubes. Unnecessary salpingectomies should, of course, not be performed and they may easily be avoided by appropriate evaluation of the tubal mucosa at laparoscopy before any final decision of salpingectomy is made. We do not see a major conflict: functional surgery to the tubes with healthy mucosa and salpingectomy to the dilated tubes that are visible on ultrasound and have a severely damaged mucosa. PMID- 11006176 TI - Effect of different cyclical sequential progestins on endometrial vascularity in postmenopausal women compared with the natural cycle: a morphometric analysis. AB - The effect of trimegestone-based and norethisterone-based hormone replacement therapy (HRT) regimens on the endometrial vascularity compared with that of the endometrium of the natural cycle were evaluated using immunohistochemical techniques. Endometrial vascular space area, diameter and number were defined in the functionalis layer of the endometrial samples from postmenopausal women who either completed a randomized, double blind, dose-ranging study of continuous oral micronized oestradiol 2 mg daily with trimegestone 0.05, 0.1, 0.25 and 0.5 mg/day from day 15-28 for six treatment cycles or were given cyclical sequential norethisterone (NET)-based HRT together with continuous 2 mg oral oestradiol valerate for three cycles. The control samples were LH-dated endometrial biopsies. NET-based HRT was associated with a higher number of smaller vascular spaces compared with the trimegestone-treated endometrium or that of the natural cycle. There was no dose-dependent effect in the four dose groups of trimegestone. In conclusion, norethisterone may exert a different effect on angiogenesis to that of trimegestone on endometrial vascular development. PMID- 11006177 TI - Serum oestradiol and progesterone concentrations during prolonged coasting in 15 women at risk of ovarian hyperstimulation syndrome following ovarian stimulation for assisted reproduction treatment. AB - Serum oestradiol and progesterone concentrations were examined for up to 7 days after withholding gonadotrophins whilst continuing pituitary down-regulation in 15 women at serious risk of severe ovarian hyperstimulation syndrome (OHSS) (serum oestradiol >6000 pg/ml and >15 follicles per ovary). Serum oestradiol concentrations rose on day 1 of coasting in all but two of the 15 women before falling, the decrement being in the order of 40% on each day. This observation permits a rational basis for the estimation of frequency of serum oestradiol measurements and duration of coasting. The trends and rates of fall of serum oestradiol do not seem to predict the occurrence of moderate and severe OHSS, being similar in the six women who developed OHSS compared with nine women without OHSS. The trends in progesterone concentrations were unrelated to any aspects of the clinical outcome. PMID- 11006178 TI - Effects of alendronate and hormone replacement therapy, alone or in combination, on bone mass in postmenopausal women with osteoporosis: a prospective, randomized study. AB - The effectiveness of hormone replacement therapy (HRT) and alendronate, alone and in combination, was evaluated in 120 postmenopausal patients with osteoporosis with bone mineral density (BMD) measurements at least 2 SD below the mean value for young premenopausal subjects. They had no contra-indications to HRT or alendronate use and were randomized to three different treatment groups. Group I was treated with micronized 17 beta-oestradiol 2 mg and norethisterone acetate 1 mg/day per os, group II received alendronate 10 mg/day per os and group III received micronized 17 beta-oestradiol 2 mg, norethisterone acetate 1 mg/day per os and alendronate 10 mg/day per os for 1 year. Elementary calcium 1500 mg/day was supplied to patients in all three groups. Spinal and femoral neck BMD and markers of bone mineral metabolism were measured on each patient before treatment and 6 and 12 months after treatment in 95 patients. At the end of the 12th month, significant increases in spinal and femoral neck BMD were found in all groups. Increases in spinal BMD were significantly higher in patients treated with alendronate and alendronate with HRT when compared with patients treated with HRT only. No significant difference was found in femoral neck BMD changes between the groups. Significant decreases in bone resorption and markers of bone formation were observed in all groups. Alendronate was found to be more effective than HRT and could have a very beneficial effect when added to the HRT regimen in patients with postmenopausal osteoporosis. Alendronate might also be used in postmenopausal patients with osteoporosis when HRT is contra-indicated or when there is reluctance to use hormonal treatment. PMID- 11006179 TI - Prospective randomized trial to evaluate the efficacy of a vaginal ring releasing progesterone for IVF and oocyte donation. AB - A polysyloxane vaginal ring containing 1g of natural progesterone was developed as luteal supplementation for women treated with IVF-embryo transfer and for agonadal women participating in an oocyte donation programme. The ring provides continuous release of progesterone (10-20 nmol/l) for 90 days. The efficacy of this form of progesterone supplementation was evaluated in two multicentre prospective randomized trials. IVF-embryo transfer trial: After oocyte aspiration, 505 women were randomly allocated to progesterone supplementation with vaginal ring or i.m. progesterone (50 mg/day). The clinical pregnancy rate was 36.6% in both groups. Implantation rate was 15.9% in the vaginal ring and 16.0% in i.m. progesterone. Oocyte donation trial: After endometrial proliferation with micronized oestradiol, 153 women were allocated to progesterone replacement with a vaginal ring or i.m. progesterone (100 mg/day). Clinical pregnancy rate was 39.8 and 28.6% respectively. Implantation rate was significantly higher with the vaginal ring compared with i.m. progesterone (19.9 and 11.6% respectively, P = 0.006). The vaginal ring is a novel development which provides continuous release of progesterone for 90 days. In IVF-embryo transfer, its effectiveness is similar to daily i.m. injections. In oocyte donation the ring provides a progestative milieu which improves the implantation rate and eliminates the discomfort of daily i.m. injections. PMID- 11006180 TI - Prediction models for insulin resistance in the polycystic ovary syndrome. AB - Women with the polycystic ovary syndrome (PCOS) have a high prevalence of insulin resistance, with consequent increased risk of metabolic diseases later in life. An early metabolic screening would therefore be of clinical relevance. By using stepwise regression analysis on several variables obtained in 72 women with PCOS, we constructed simple and reliable mathematical models predicting insulin sensitivity, as measured by the euglycaemic hyperinsulinaemic clamp. The normal ranges of insulin sensitivity were calculated from 81 non-hirsute, normally menstruating women with normal ovaries, and similar body mass index (BMI) and age as the women with PCOS. Measured variables included BMI, waist and hip circumferences, truncal-abdominal skin folds, circulating concentrations of gonadotrophins, androgens, sex hormone-binding globulin (SHBG), triglycerides, total cholesterol and cholesterol subfractions, fasting insulin, C-peptide and free fatty acids. The three best prediction models included waist circumference, together with insulin (model I: R(2) = 0.77), serum triglycerides (model II: R(2) = 0.65), and the subscapularis skin fold (model III: R(2) = 0. 64). Using reference limits for insulin sensitivity obtained in the 81 normal pre-menopausal women, the models identify insulin resistant women with PCOS. These simple and inexpensive models are potentially useful in clinical practice as an early screening in women with PCOS. PMID- 11006181 TI - Reproductive hormones, cancers, and conditions in relation to a common genetic variant of luteinizing hormone. AB - A variant of the beta-subunit of luteinizing hormone (v-LH) is more common among populations also at higher risk for breast and ovarian cancer. To explore the possible relationship between these cancers and v-LH, we examined its frequency in premenopausal women, including 100 with a family history of ovarian cancer, 94 with carcinoma- in-situ of the breast, and 153 age and residence-matched controls. Reproductive histories were assessed and v-LH status measured by immunological assays from plasma drawn during the early follicular phase of cycles. For the entire study population, 283 (81.5%) were wild type; 61 (17.6%) were heterozygous; and three (0.9%) were homozygous for v-LH. Carrier frequency was not elevated among women with a family history of ovarian cancer or personal history of carcinoma-in-situ of the breast compared with controls. Women with the v-LH variant were less likely to report menstrual weight gain or ovarian cysts, more likely to report infertility, and have higher early follicular phase LH concentrations compared with women who were wild type. While there is no evidence from this study that v-LH increases risk for ovarian or breast cancer, we conclude that possession of v-LH may impact on some aspects of reproductive history and LH concentrations. PMID- 11006182 TI - Why are boys more likely to be preterm than girls? Plus other related conundrums in human reproduction. AB - The present note offers two quite different points of interest. The first concerns the cause(s) of preterm labour. This is an important practical problem because of the associated morbidity and mortality. The second point of interest is theoretical. Some logically inter-related empirical propositions are formulated: none has been established but evidence for each is apparently evidence for all. However, as far as I know, there is no formal method of treating such material. It would be useful if methodologists would offer an opinion on whether the practical problem of the causes of preterm birth could be helpfully approached by the accumulation of data on the propositions outlined here. In my opinion, critics should not assess these propositions piecemeal, but should consider the whole edifice of propositions simultaneously. Obstetricians who confine their attention to the delivery of babies should extend it to the circumstances surrounding the conception of those babies. For there may lie the solution to the problem of why boys are more likely to be preterm than girls. PMID- 11006183 TI - Hysteroscopic appearance of the mid-secretory endometrium: relationship to early phase pregnancy outcome after implantation. AB - A total of 172 patients who underwent hysteroscopic assessment of the endometrium and then became pregnant, was analysed retrospectively to explore the relationship between endoscopic findings and early phase pregnancy outcome after implantation. Histological examination of the endometrium and assay of serum progesterone and oestrogen were carried out simultaneously with hysteroscopy. Of 172 patients, 12 were excluded. Of the remaining 160 patients, 62 (38.8%) were classified endoscopically as having 'good' mid-secretory endometrium and 98 (61.3%) as 'poor', between one and four cycles prior to the conception cycle. There were no clinical differences between these two groups, except that the frequency of patients with a history of early abortion was significantly higher in the 'poor' group (25.5%) than in the 'good' group (8.1%) (P < 0.05). Of 160 pregnancies, 118 persisted successfully to live birth, but 42 ended in early pregnancy loss. The incidence of early abortion was significantly higher in the 'poor' group (33.7%) than in the 'good' group (14.5%) (P < 0.05). Significant differences were observed between the two groups for histological dating of the endometrium (P < 0.05) but not for serum progesterone and oestradiol concentrations or progesterone:oestradiol ratio. In conclusion, our data suggest that the hysteroscopic appearance of the mid-secretory endometrium at this stage of the menstrual cycle is a better prognostic factor for pregnancy outcome than hormonal data. PMID- 11006184 TI - Different patterns of structural luteolysis in the human corpus luteum of menstruation. AB - Structural luteolysis is a complex process responsible for the elimination of the corpus luteum (CL). The aim of this study was to analyse the luteolytic process of the CL of menstruation. For this, we have morphologically studied 654 ovaries from 340 cycling women. Apoptotic cells were observed almost exclusively during the perimenstrual period and were extremely scarce at advanced stages of involution. Steroidogenic luteal cells surviving to the perimenstrual apoptotic wave underwent characteristic degenerative changes, consisting of intense cytoplasmic vacuolation, expression of macrophage markers and accumulation of lipofuscin pigment, and they persisted for long periods of time. Accumulation of corpora albicantia (CA) was observed in only 25% of a subset of 168 women, whereas 28% showed involuting CL without hyalinization, consisting of clusters of pigment-filled cells, and 46.4% showed ovaries with a mixture of CA and involuting CL without hyalinization or involuting CL with intermediate features. Evolution of the CL towards CA seemed to be related to the presence of a large, blood-filled cavity. The data from this study suggested that different patterns of structural luteolysis exist during CL involution, and that the final fate of the involuting CL is dependent on the presence of a large, central, blood-filled cavity. PMID- 11006185 TI - Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders: a case series. AB - In patients with poor response to ovarian stimulation with gonadotrophins, growth hormone (GH) is sometimes used to increase paracrine insulin-like growth factor-1 (IGF-1) effect. We postulated that dehydroepiandrosterone (DHEA) administration to poor responders would augment gonado-trophin effect via a similar mechanism. Baseline ovarian stimulation response to a cycle with DHEA in five healthy non smoking women <41 years old was compared with day 3 FSH <20 mIU/ml. All had documented poor response to vigorous gonadotrophin administration. After day 2 ultrasounds, DHEA-sulphate (DHEA-S), FSH, human chorionic gonadotrophin (HCG), and testosterone were measured, and the women were given 80 mg/day of oral micronized DHEA for 2 months. While still on DHEA, they underwent ovarian stimulation with FSH given i.m. twice a day, and HCG (10 000 IU) at follicular maturity, followed by intrauterine insemination. Cycle parameters assessed were peak oestradiol, and peak oestradiol/ampoule. The DHEA/ovarian stimulation cycles occurred between 4 and 24 months after the control cycles. After 2 months DHEA treatment, DHEA-S increased to 544 +/- 55 microg/dl, and testosterone increased to 67.3 +/- 6.1 ng/dl. All five subjects (six cycles; one subject had two DHEA cycles) had increased responsiveness; peak oestradiol concentrations increased from 266.3 +/- 69.4 pg/ml to 939.8 +/- 418.9 pg/ml. The oestradiol/ampoule ratio increased in all six cycles, by a mean of 2.94 +/- 0.50 fold (P = 0.012). One of the cycles resulted in a delivered twin pregnancy. In this small series, DHEA improved response to ovarian stimulation even after controlling for gonadotrophin dose. Supplemental DHEA treatment during ovarian stimulation may represent a novel way to maximize ovarian response. PMID- 11006186 TI - Oocyte and semen donation: a survey of UK licensed centres. AB - In view of the difficulties encountered by licensed clinics in the recruitment of oocyte and semen donors, the present survey was commissioned by the National Gamete Donation Trust (NGDT) to provide systematic information on current practice as a basis for planning future recruitment strategies. Sixty-four clinics agreed to participate in the survey, of which 55 recruited oocyte donors, 30 recruited semen donors, and 24 recruited both oocyte and semen donors. Almost all of the clinics had experienced difficulty in obtaining a sufficient supply of donated oocytes, and many patients requiring treatment with donated oocytes were placed on a waiting list for more than one year. Three-quarters of potential donors changed their mind about donating after receiving information on the procedures involved. The difficulties in recruiting semen donors were generally less acute. In this case, the greatest problem was the high proportion of potential semen donors who were deemed unsuitable after completing the screening process. Clinics also reported a shortage of both oocyte and semen donors from specific ethnic groups. PMID- 11006187 TI - The outcome of IVF-embryo transfer treatment in patients who develop three follicles or less. AB - Among 828 patients undergoing IVF-embryo transfer treatment, the implantation and pregnancy rates of patients who developed < or = 3 follicles were compared prospectively with those patients who had a normal response. Patients who developed 1 to 3 follicles during ovarian stimulation elected to proceed with oocyte collection, have intrauterine insemination if appropriate, or to have their cycle cancelled. In the group of patients who developed < or = 3 follicles and who were aged <40 years, despite a significantly lower number of oocytes collected [2 versus 7; median difference (MD) = 9; confidence interval (CI) = 7 11, and lower number of embryos developed and transferred (1 versus 3; MD = 2; CI = 1-2), no difference in either implantation rate [27.8 versus 20.4%; odds ratio (OR) = 1.58; CI = 0.46-4.54] or pregnancy rate (27.8 versus 36.7%; OR = 0.7; CI = 0.2-2.0) was noted when compared with similarly aged patients who developed >3 follicles. However, in patients aged >40 years who developed < or = 3 follicles, a moderate, albeit non-significant decrease in implantation rate (3.8 versus 7.8%; OR = 1.91; CI = 0.4-57.0) and pregnancy rate (4.2 versus 18.3%; OR = 1.92; CI = 0.38-57.0) was observed when compared with patients of a similar age who developed >3 follicles. Patients aged <40 years, unlike older patients, maintain good implantation and pregnancy rates despite a poor response to ovarian stimulation. This study indicates that for this group of women, continuation of IVF treatment is a better option than cancellation. PMID- 11006188 TI - Addition of GnRH antagonist in cycles of poor responders undergoing IVF. AB - Concern about the use of gonadotrophin-releasing hormone (GnRH) agonists in ovarian stimulation of poor responder IVF patients has arisen from the claim that GnRH agonists might have a direct deleterious effect through their receptors on the ovary. In this study, we compared two ovarian stimulation protocols in which no GnRH agonists were used. In all, 40 patients with a poor response in previous treatment cycles were included. They were divided into two groups: group I (n = 20) received ovarian stimulation for 20 cycles, without the addition of either GnRH agonist or antagonist; while group II (n = 20) patients received ovarian stimulation for 20 cycles, including the administration of a GnRH antagonist (Cetrorelix, 0.25 mg daily) during the late follicular phase. There was no statistically significant difference between the groups for mean age, duration of infertility, baseline FSH concentration, cancellation rate, number of ampoules of gonadotrophin used, number of mature oocytes retrieved, oestradiol concentrations on the day of injection of human chorionic gonadotrophin (HCG), fertilization rate and number of embryos transferred. The clinical pregnancy and implantation rates in group II appeared higher than in group I, but were not significantly different (20 and 13.33% compared with 6.25 and 3.44% respectively). The addition of GnRH antagonists to ovarian stimulation protocols might be a new hope for poor responder IVF patients, but this report is preliminary and further controlled randomized prospective studies with larger sample sizes are required. PMID- 11006189 TI - Comparison of two different doses of lignocaine used in paracervical block during oocyte collection in an IVF programme. AB - We have recently demonstrated the efficacy of paracervical block (PCB) used in conjunction with conscious sedation during egg collection. The dosage of lignocaine used in various studies ranges from 50 mg to 200 mg. There are, however, no studies evaluating the efficacy of different doses of local anaesthetic agents used in PCB. In this prospective, double-blind and placebo controlled study, 150 women undergoing egg collection in their first IVF cycle were randomized to receive 200 mg and 150 mg lignocaine in PCB. No differences were seen in the demographic data, the ovarian responses, the duration of egg collection and the number of follicles punctured. The fertilization, implantation and pregnancy rates were similar when either 150 mg or 200 mg lignocaine was employed. The median pain levels during vaginal punctures were 14.0 (2.5th-97.5th centiles: 0-75.4) and 14.0 (2.5th-97.5th centiles: 0-86.5) in patients receiving 200 mg and 150 mg lignocaine respectively, whereas the corresponding median abdominal pain levels were 14.0 (2.5th-97.5th centiles: 0-85.6) and 14.0 (2.5th 97.5th centiles: 0-99.1). These pain levels during egg collection were not significantly different between the two groups. The use of 200 mg lignocaine in PCB is not justified, even in the absence of toxic effects. PMID- 11006190 TI - Successful pregnancy and birth after IVF in a woman with cystic fibrosis. AB - As survival increases, patients with cystic fibrosis (CF) are often confronted with reproductive issues. Initial reports gave conflicting advice regarding the outcome of pregnancy in CF. However a recent large longitudinal study of pregnancies in CF women suggested that pregnancy has little impact on morbidity or mortality. Reduced fertility in CF women has been described, possibly due to thickened cervical mucus, and intrauterine insemination (IUI) has been used to overcome this. We report the first woman with CF, to our knowledge, to be successfully treated with IVF after repeated failed attempts at IUI. PMID- 11006191 TI - Fertility preservation of boys undergoing anti-cancer therapy: a review of the existing situation and prospects for the future. AB - With the advancement of medical science, most cancers in children are now treatable, the cure rate being almost 85%. In boys, one side effect of treatment (chemotherapy and radiotherapy) is destruction of the sperm precursor cells in the testis, resulting in the failure of sperm formation after puberty, and consequent infertility. At the time of anti-cancer treatment, future fertility of the boy patient is at the very bottom of the relative quality of life (RQL) parameters list; however, in adults infertility is an important issue following cure from cancer. In this article we have first reviewed the existing situation with regard to the state of the art of fertility preservation in young boys with cancer, and have then raised clinical and ethical issues and suggested a way forward. The review concludes with the emphasis that certain important issues still need to be resolved and, until these are, then the different techniques available should be confined to approved, ethical clinical trials where efficacy and safety can be fully evaluated. PMID- 11006192 TI - Membrane fluidity predicts the outcome of cryopreservation of human spermatozoa. AB - Semen cryopreservation is an important procedure in the treatment of human infertility. However, the ability of spermatozoa to survive freeze/thaw processes varies between patients. Cryopreservation-induced stress may result in membrane injury with consequent loss of sperm motility and viability. We investigated the relationship between the physico-chemical state of the human sperm membranes and their tolerance to cryopreservation. Conventional characteristics of 20 semen samples were analysed before and after cryopreservation as well as their membrane fluidity assessed by measuring the fluorescence polarization anisotropy, which is inversely proportional to the fluidity. Correlation between fluidity and post thaw recoveries of motile and viable spermatozoa were examined. Results showed that membrane anisotropy markedly varies between patients. In cryopreserved spermatozoa, anisotropy values were significantly higher than in fresh spermatozoa. Furthermore, recovery of motile and viable spermatozoa after freeze/thaw was strongly correlated with anisotropy of fresh spermatozoa (P < 0.05). The higher the membrane fluidity was before freezing, the better was the response of spermatozoa to cryopreservation. The results indicate that the freeze/thaw process results in a rigidifying effect on the sperm membrane and suggest that sperm adaptability to freeze/thaw-induced stress could be dependent on their initial membrane fluidity. The latter finding has practical implications for predicting the response of spermatozoa following freezing and thawing and for improving the recovery of viable spermatozoa. PMID- 11006193 TI - Frequency of hyper-, hypohaploidy and diploidy in ejaculate, epididymal and testicular germ cells of infertile patients. AB - The hypothesis that sperm aneuploidy and diploidy increase as a function of spermatogenesis impairment was addressed. Ejaculated semen samples from a series of men (n = 22) with very low total normal motile count (1 x 10(6)) was analysed in terms of sperm aneuploidy and diploidy by in-situ hybridization and compared with controls (n = 10). Germ cell aneuploidy was also analysed in an additional series of infertile patients presenting unexplained infertility (n = 3), congenital absence of the vas deferens (CAVD) (n = 6) and non-obstructive azoospermia (n = 3) undergoing IVF, microsurgical epididymal sperm aspiration (MESA)/ICSI and testicular sperm extraction (TESE)/ICSI cycles respectively. In situ hybridization for chromosomes 1, 17, X and Y was performed on ejaculate, epididymal and testicular spermatozoa. Significantly higher sperm aneuploidy and diploidy rates where found (for the four chromosomes analysed) in spermatozoa from oligoasthenoteratozoospermia (OAT) over controls (18 versus 2.28% and 2.8 versus 0.13% respectively; P < 0.001). Testicular germ cells had even higher rates of sperm aneuploidy and diploidy. However, in this group it was difficult to determine whether the cells analysed were dysmorphic spermatozoa or spermatids. The data warrant further investigation on the cytogenetic abnormalities found in most germ cells identified in testicular tissue biopsies of azoospermic patients. PMID- 11006194 TI - Improvement in motion characteristics and acrosome status in cryopreserved human spermatozoa by swim-up processing before freezing. AB - The purpose of this study was to examine if selecting a sperm population with improved motion characteristics before freezing reduces the deleterious effects of cryopreservation. Semen specimens from 15 normal donors were divided into two equal aliquots. The first aliquot received no treatment (control), and the second was processed by swim-up from a washed sperm preparation to select a sperm population with better motility and motion characteristics (swim-up). Both aliquots were cryopreserved by the liquid nitrogen vapour method. Percentage motility and motion characteristics were evaluated by computer-assisted semen analysis. Acrosome integrity as well as spontaneous and calcium ionophore-induced acrosome reactions before freezing and after thawing were assessed by fluorescein isothiocyanate conjugated peanut agglutinin combined with a supra vital dye (Hoechst-33258). Swim-up processing enabled selection of a sperm population with better motion characteristics, percentage motility and viability before freezing (P < 0.001), but with no difference in percentage of acrosome-intact spermatozoa (P = 0.63). After thawing, the swim-up specimens exhibited faster velocity and progression than untreated specimens (P < 0.001). They also had higher percentages of spermatozoa with intact acrosomes and spermatozoa able to undergo acrosome reaction in response to calcium ionophore (P < 0.05). Selecting a highly motile sperm population before freezing enhances overall post-thaw spermatozoa quality. PMID- 11006195 TI - Differential expression and localization of de-novo synthesized endometriotic haptoglobin in endometrium and endometriotic lesions. AB - Endometriosis protein-I (ENDO-I) mRNA expression and protein localization were evaluated using in-situ hybridization and immunohistochemistry in endometriotic lesions and eutopic endometrium from women with endometriosis, and in eutopic endometrium from women without endometriosis (controls). When present, ENDO-I mRNA and protein were observed in the functionalis zone of endometrial stroma and the stroma of endometriotic lesions. Expression and localization differences were scored and statistically analysed. During the secretory stage, ENDO-I mRNA expression by endometriotic lesions and eutopic endometrium from women with disease was significantly greater than ENDO-I mRNA expression by proliferative stage eutopic endometrium from women with disease or eutopic endometrium from controls, regardless of cycle stage (P < 0.001). More ENDO-I protein was localized in endometriotic lesions and eutopic endometrium from women with disease than in eutopic endometrium from controls, regardless of cycle stage (P < 0.001). Differential expression and localization of ENDO-I may help develop minimally invasive diagnostic strategies for endometriosis. Further, as ENDO-I shares nucleotide sequence and amino acid sequence with hepatic haptoglobin-which in certain disease states is immunosuppressive and angiogenic-differences in ENDO I expression and localization in the peritoneal cavity may contribute to the pathogenesis of endometriosis and/or facilitate development of unprecedented diagnostic or therapeutic approaches for management of this enigmatic disease. PMID- 11006196 TI - Safety of cryopreservation straws for human gametes or embryos: a preliminary study with human immunodeficiency virus-1. AB - The aim of this preliminary experimental study was to test the stability of cryopreservation straws to human immunodeficiency virus-1 (HIV-1). Three kinds of straws were tested: four polyvinyl chloride (PVC), four polyethylene terephthalate glycol (PETG) and 20 high-security ionomeric resin (IR). The PVC and PETG straws were sealed ultrasonically, and the IR straw by thermosoldering. Each sealed straw was cut in half to produce two demi-straws and then filled with 100 microl of HIV-1-containing supernatant (reverse transcriptase activity: 15 000 c.p.m./50 microl). The unsealed cotton end of PVC and PETG straws and the two halves of the IR straws (cotton and plastic plug ends) were tested. Each demi straw was two- thirds submerged in RPMI medium at 37 degrees C, and RPMI samples were withdrawn on days 3, 7 and 11. Viral RNA was extracted from the medium and then amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) followed by nested PCR using primers specific to HIV-1 protease. On day 7, no HIV 1 RNA was detected in any of the different samples of medium that had surrounded the unsealed PVC and PETG straws with cotton ends, but three IR specimens were positive. On day 11, PVC and PETG remained negative but HIV-1 RNA was detected in RPMI samples for two more IR demi-straws (n = 5). In conclusion, under these experimental conditions (at 37 degrees C), the unsealed cotton end PVC, PETG and thermosoldered cotton end IR demi-straws appeared to be safe for HIV-1, while IR straws, sealed or unsealed with a plastic plug and with unsealed cotton ends, leaked. PMID- 11006197 TI - Morphological evaluation of human embryos and derivation of an embryo quality scoring system specific for day 3 embryos: a preliminary study. AB - A scoring system specific for day 3 embryos has not been extensively explored. Most IVF laboratories continue to grade embryos solely on the basis of cell number and percentage fragmentation as was traditionally done for day 2 embryos. Additional morphological features, some unique to day 3 embryos, may be useful in selecting embryos most likely to blastulate and implant. The objective of this study was to derive an embryo scoring system for day 3 transfers which is predictive of positive pregnancy outcomes. A total of 316 transferred embryos from 93 patients was recorded on videotape and evaluated. The following parameters were used to grade the embryos: cell number, fragmentation pattern (FP), cytoplasmic pitting, compaction, equal sized blastomeres, blastomere expansion and absence of vacuoles. The clinical pregnancy rate was 41.9%, with an implantation rate of 18% per embryo transferred. The mean number of embryos transferred per patient was 3.4. Three formulae were derived to score embryo quality in each transfer based on the average score of individual embryos transferred. In the first scoring system, cell number alone was used to predict pregnancy outcome. The second scoring system was based on blastomere number and the observed FP. The third scoring system utilized both blastomere number and FP but also combined this with five morphological criteria to yield a final day 3 embryo quality (D3EQ) score. We found the D3EQ score to be prognostic of pregnancy outcome. This study suggests that although cell number and FP are certainly predictors of positive pregnancy outcomes, additional parameters specific to day 3 embryos should be used to stratify a cohort of embryos further. PMID- 11006198 TI - The effect of propofol anaesthesia on oocyte fertilization and early embryo quality. AB - Propofol, frequently used for i.v. induction of anaesthesia in assisted reproduction procedures, has been suspected of damaging oocytes. Concentrations of propofol have recently been shown to increase in follicular fluid during oocyte retrieval. Our study was designed to assess whether exposure to increasing concentrations of propofol has a measurable effect on in-vitro fertilization, cleavage and embryo development. A cohort of 130 women underwent i.v. anaesthesia using propofol and fentanyl. Time of anaesthesia from i. v. injection of propofol was measured, as were the doses of the two drugs. In 32 women expected to have more than 15 oocytes retrieved, first, middle and last oocytes were cultured separately. The mean time from i.v. injection to first follicle aspiration was 200 s. The mean time for the aspiration of each additional oocyte was 17.6 s. In 10 out of 11 cases where follicular fluid concentrations of propofol were measured, there was an increase from the first to the last follicle, but no difference was found in the ratio of mature to immature oocytes. Nor were any differences found in fertilization, cleavage and embryo cell number. In so far as in-vitro development reflects embryo quality, we conclude that the time elapsed between retrieval of the first and last oocyte does not affect oocyte quality. PMID- 11006199 TI - The use of whole rat embryo culture as a technique for investigating potential serum toxicity in recurrent miscarriage patients. AB - Previously, the rat embryo model has been used as an experimental technique in investigations of the aetiology of idiopathic recurrent miscarriage. The aim of the present study was to validate it as a tool in the investigation of the aetiology of this condition. Subjects (n = 36) with a history of recurrent miscarriage were recruited from two dedicated recurrent miscarriage clinics and compared with control women with at least one previous pregnancy resulting in a live birth (n = 23). Serum from each woman was used as culture medium in the rat embryo model. Cultured embryos were scored for growth and differentiation. No statistical difference was found in any parameter between the two groups. Furthermore, patients from the recurrent miscarriage group whose serum demonstrated a trend towards lower scores, subsequently conceived and underwent uncomplicated pregnancies. PMID- 11006200 TI - Randomized comparison of vaginal (200 microg every 3 h) and oral (400 microg every 3 h) misoprostol when combined with mifepristone in termination of second trimester pregnancy. AB - It is known that when misoprostol is given at 200 microg every 3 h after mifepristone pretreatment, the vaginal route is more effective than the oral route. However, women prefer the oral route. This randomized study was to test our hypothesis that oral misoprostol 400 microg is as effective as vaginal misoprostol 200 microg when given every 3 h in termination of second trimester pregnancy after priming with mifepristone. A total of 142 patients was randomly assigned to group 1 (200 mg mifepristone + 400 microg oral misoprostol every 3 h up to five doses) or group 2 (200 mg mifepristone + 200 microg vaginal misoprostol every 3 h up to five doses). The incidence of side-effects and the preference study were assessed through a standardized questionnaire during and after the abortion. For the oral group, both the incidence of diarrhoea (40.0 versus 23.2%, P = 0.03) and the amount of drug used (1734 compared with 812 microg, P < 0.0001) were significantly higher than that of the vaginal group but the incidence of fever appeared to be lower (not significant). There was no significant difference in complete abortion rate: 81.4% in the oral group and 75.4% in the vaginal group. The median induction-abortion interval was similar in the two groups (10.4 versus 10.0 h). The percentage of women who aborted in 24 h was also similar: 57/70 (81.4%) in the oral group and 58/69 (87.0%) in the vaginal group. Overall, 82.0% of women preferred the oral route. Oral misoprostol (400 microg) given every 3 h up to five doses, when combined with mifepristone, was as effective as the vaginal (200 microg) route in second trimester termination of pregnancy. This regimen could also be offered to those women who found repeated vaginal administration unacceptable. PMID- 11006201 TI - Choriocarcinoma-like human chorionic gonadotrophin (HCG) and HCG bioactivity during the first trimester of pregnancy. AB - The objective of this study was to evaluate the distribution of choriocarcinoma like human chorionic gonadotrophin (HCG) isoforms during first trimester pregnancy and their relationship with in-vitro HCG bioactivity. This was done by means of a retrospective analysis of patients' sera with first trimester normal intrauterine and abnormal (ectopic) pregnancies. Serum samples were obtained from 38 women with an amenorrhoea of <10 weeks. From these, 19 had a normal intrauterine pregnancy (IUP) and 19 an ectopic pregnancy (EP). Total immunoreactive HCG (HCGi), free beta-HCGi and oestradiol were measured by enzyme immunoassays and bioactive HCG by the mouse Leydig cell bioassay. The alterations in HCG isoform content were measured by the combination of two immunometric assays, B152 for choriocarcinoma-like HCG and B109 for intact HCG detection and expressed as the B152/B109 ratio. Choriocarcinoma-like HCG isoforms ratio measured by B152 and B109 assays was significantly higher in the low subgroups of free beta-HCGi and gestational age (P = 0.0111 and 0.0036 respectively). Whereas bioactive to immunoreactive HCG ratios (b/i ratio) were significantly higher when free beta-HCGi concentrations were low (P = 0.0010), no correlation was found between the variation of bioactivity (b/i ratio) and the proportion of choriocarcinoma-like HCG isoforms (B159/B108). It is concluded that in first trimester pregnancies (i) the modulation of HCG in-vitro bioactivity is not related to the variation of choriocarcinoma-like HCG isoforms secretion and (ii) the amount of choriocarcinoma-like HCG isoforms secreted by the early trophoblast is predominant and may be the result of an early developmental regulation of glycosylation enzyme. PMID- 11006202 TI - A longitudinal study of maternal plasma insulin-like growth factor binding protein-1 concentrations during normal pregnancy and pregnancies complicated by pre-eclampsia. AB - Insulin-like growth factor binding protein-1 (IGFBP-1) is synthesized by the decidual stroma, and is thought to act locally to inhibit IGF activity and so limit trophoblast invasion. Cross-sectional studies have reported conflicting data on maternal circulating concentrations of IGFBP-1 in early pregnancy before the development of pre-eclampsia. A longitudinal study was performed in 10 women who went on to develop pre-eclampsia and a group of 12 normal pregnant controls, chosen to be similar for maternal age, booking body mass index (BMI) and gestational age. Maternal IGFBP-1 concentrations were measured in plasma obtained at 16, 20, 24, 28, 32 and 36 weeks. Plasma IGFBP-1 concentrations were unchanged over this period in normal pregnancy. In contrast, the concentrations in women who developed pre-eclampsia increased progressively. At 16, 20, and 24 weeks the concentrations were significantly lower compared to normal pregnancy, at 28 and 32 weeks, similar, but by 36 weeks the concentrations were significantly greater than the normal controls. The data show that circulating IGFBP-1 concentrations are lower in early pregnancy before the development of pre-eclampsia. Thus, it is suggested that IGFBP-1-induced inhibition of IGF activity is unlikely to be responsible for the impaired trophoblast invasion observed in pre-eclampsia. PMID- 11006203 TI - Recurrent miscarriage--an aspirin a day? AB - Recurrent miscarriage and later pregnancy complications are in some cases associated with placental thrombosis and infarction. The aim of this study was to assess the value of low dose aspirin (75 mg daily) in improving the subsequent livebirth rate amongst women with either unexplained recurrent early miscarriage (<13 weeks gestation; n = 805) or unexplained late pregnancy loss (n = 250). Amongst women with recurrent early miscarriages, there was no significant difference in the livebirth rate between those who took aspirin (251/367; 68.4%) compared with those who did not take aspirin [278/438; 63.5%; odds ratio (OR) 1.24; 95% confidence interval (CI) 0.93-1.67]. This relationship was independent of the number of previous early miscarriages. In contrast, women with a previous late miscarriage who took aspirin had a significantly higher livebirth rate (122/189; 64.6%) compared with those who did not take aspirin (30/61; 49.2%: OR 1.88; 95% CI 1.04-3.37). The empirical use of low dose aspirin amongst women with unexplained recurrent early miscarriage is not justified. We are currently investigating the role of incremental doses of aspirin in the treatment of women both with early miscarriages associated with thrombophilic abnormalities and in those with late pregnancy losses. PMID- 11006204 TI - Chemical ripening of the cervix with intracervical application of sodium nitroprusside: a randomized controlled trial. AB - Nitric oxide (NO) has been found to be involved in the processes of cervical ripening. In a randomized, placebo-controlled study, cervical softening by an intracervical application of sodium nitroprusside, one of the most clinically potent and effective NO donor agents, was evaluated. A total of 36 primigravid women undergoing pregnancy termination between 9 and 12.5 weeks were enrolled. In one series, 18 patients were randomized to receive intracervically either placebo or 1% nitroprusside gel (5 mg), followed by uterine evacuation 6 h after treatment. In another series, 18 patients received either placebo or 2% nitroprusside gel (10 mg) into the cervical canal followed by uterine evacuation 3 h later. The cervical resistance, i.e. the force required to dilate the cervix from 3 to 10 mm, was the main outcome variable. It was recorded using a force sensing apparatus (dynamometer). Blood pressure was measured. Adverse events were recorded until 2 h after surgery. Women treated with both doses of nitroprusside gel showed values of cervical resistance significantly lower than those treated with placebo gel, at any tested diameter. No differences were found between subjects treated with the two different doses of nitroprusside. No significant consistent changes in blood pressure were induced by either dose of nitroprusside. No headaches were found in subjects treated with the NO donor. This study demonstrates that sodium nitroprusside applied into the cervical canal induces a rapid and significant softening of the cervix, thus reducing the force required to dilate it, compared with placebo-treated subjects. The chemical ripening of the cervix with sodium nitroprusside intracervical gel is an efficacious procedure in first-trimester pregnancy. PMID- 11006205 TI - Embryo reduction of multifetal pregnancies following assisted reproduction treatment: a modification of the transvaginal ultrasound-guided technique. AB - First-trimester transvaginal embryo reduction is an effective alternative for the management of multifetal pregnancy in assisted reproduction. We have modified the transvaginal technique by performing an intracardiac embryo puncture until asystolia is verified, without the injection of any substances. Any aspiration of embryo tissues or amniotic fluid was avoided. A total of 149 multifetal pregnancies was reduced to twins (n = 134) or singletons (n = 15) at early gestational age (7.8 +/- 0.8 weeks). Eleven cases (7.3%) of miscarriage, two cases (1.3%) of chorioamnionitis, and 17 cases (11.4%) of transient spotting were recorded as postoperative complications. Vanishing of one embryo occurred in four cases (3.0%) of those reduced to twins. The baby take-home rate was 89.5% for twins and 80.0% for singletons. Pregnancy outcome was analysed and compared with a control group of women with non-reduced multiple pregnancies. The birth weight of singleton pregnancies after reduction was lower (2929 +/- 160 versus 3291 +/- 422 g; P < 0.02). These studies show that early transvaginal intracardiac embryo puncture is an effective and safe technique. PMID- 11006206 TI - Chorioamnionitis decreased incidence of respiratory distress syndrome by elevating fetal interleukin-6 serum concentration. AB - Respiratory distress syndrome (RDS) of newborns is one of the most important factors determining neonatal morbidity and mortality. The interleukin-6 (IL-6) titre in cord sera of RDS-free neonates born to mothers with histological chorioamnionitis was significantly higher than that in RDS-complicated neonates without chorioamnionitis. Maternal administration of glucocorticoid suppressed the IL-6 concentrations in the cord sera of fetuses with chorioamnionitis. The fetuses without chorioamnionitis who suffered from RDS even after maternal glucocorticoid administration showed a similar IL-6 titre to that of RDS-affected neonates without chorioamnionitis. Examination of the mechanism by which IL-6 decreased the incidence of fetal RDS revealed that H441-4, a human pulmonary adenocarcinoma cell line, stimulated with recombinant (r)-IL-6 started the synthesis of mRNA and protein of pulmonary surfactant protein (SP)-A. The present study shows that IL-6 elevation in fetuses with chorioamnionitis promotes fetal lung maturation by inducing SP-A synthesis, thereby decreasing the incidence of RDS in the preterm neonates. PMID- 11006207 TI - ESHRE guidelines for good practice in IVF laboratories. Committee of the Special Interest Group on Embryology of the European Society of Human Reproduction and Embryology. AB - Education has always been a priority for the European Society of Human Reproduction and Embryology (ESHRE). Many efforts have been dedicated to promoting knowledge of techniques, procedures and strategies in order to ensure use of the highest quality practices in reproductive medicine. The need to develop a set of guidelines was a logical consequence that found its first expression in 1990, when Focus on Reproduction (vol. 1, pp. 10-38) published the first guidelines which were distributed among the membership. Five years later a new, more complete edition with several novel techniques and developments appeared in Human Reproduction (vol. 10, pp. 1246-1271). Both have proved to be invaluable references. Five more years have now passed. The necessity to produce current guidelines for good IVF laboratory practice has provided the strongest motivation. This originated from the increasing awareness that embryologists have a duty to prevent unintentional incidents that might result from poor practice in the laboratory. Therefore, the Embryology Special Interest Group (SIG) undertook to draw up guidelines aimed at giving support and guidance to the laboratory staff. All the aspects required to provide a safe working system were taken into consideration by members of the SIG and their effort produced this document. We hope that it will assist staff in achieving the best clinical outcome for their patients. PMID- 11006208 TI - Recruitment of only cytomegalovirus (CMV) negative semen donors. PMID- 11006209 TI - The double-edged ocular immune response: the Cogan lecture. PMID- 11006210 TI - Adipogenesis in thyroid eye disease. AB - PURPOSE: Adipogenesis contributes to the pathogenesis of thyroid eye disease (TED). Thyrotropin receptor (TSHR) transcripts are present in orbital fat. This study was conducted to determine whether they are expressed as functional protein, and if so, whether this is restricted to TED orbits or to a particular stage in adipocyte differentiation. METHODS: Samples of fat were obtained from 18 TED-affected orbits and 4 normal orbits, and 9 were obtained from nonorbital locations. Frozen sections were examined by immunocytochemistry using monoclonal antibodies specific for the human TSHR. Samples were disaggregated and the preadipocytes separated from the mature by differential centrifugation and cultured in serum-free or DM and examined for morphologic changes, oil red O and TSHR staining, and TSH-induced cyclic adenosine monophosphate (cAMP) production. RESULTS: Marked immunoreactivity was observed in frozen sections from all three TED samples and faint staining in both normal orbital fat samples. In vitro, 1% to 5% of preadipocytes displayed TSHR immunoreactivity in five of six TED and two of three normal orbital samples and in three of five nonorbital samples. Differentiation, was induced in all 14 orbital samples. Three of four nonorbital samples contained occasional differentiated cells. Fifty percent to 70% of differentiating cells demonstrated receptor immunoreactivity. Two of three TED and four of four nonorbital preadipocytes in DM and/or mature adipocytes displayed a TSH-mediated increase in cAMP. CONCLUSIONS: The results indicate that orbital fat TSHR transcripts are expressed as protein, which can be functional. This is not aberrant in TED orbits, although expression may be upregulated. The majority of preadipocytes undergoing differentiation express the receptor, indicating a key role for this population in one mechanism for increasing orbital volume. PMID- 11006211 TI - MRI dynamic color mapping: a new quantitative technique for imaging soft tissue motion in the orbit. AB - PURPOSE: To investigate both feasibility and clinical potential of magnetic resonance imaging-dynamic color mapping (MRI-DCM) in measuring the motion of soft tissues in the orbit and in the diagnosis of orbital disorders by detecting changes in motion. METHODS: Sequences of MRI scans were acquired (acquisition time, 5 seconds) in a shoot-stop manner, while the patient fixated at a sequence of 13 gaze positions (8 degrees intervals). Motion was quantified off-line (in millimeters per degree of gaze change) using an optical flow algorithm. The motion was displayed in a color-coded image in which color saturation of a pixel shows the displacement and the hue the displacement's orientation. Six healthy volunteers and four patients (two with an orbital mass and two with acrylic ball implant after enucleation) were studied. RESULTS: The technique was found to be clinically feasible. For a gaze change of 1 degrees, orbital tissues moved between 0.0 and 0.25 mm/deg, depending on the type of tissue and location in the orbit. In the patients with an orbital mass, motion of the mass was similar to that of the medial rectus muscle, suggesting disease of muscular origin. In the enucleated orbits, soft tissue motion was decreased. One eye showed attachment of the optic nerve to the implant, which could be verified by biopsy. CONCLUSIONS: MRI-DCM allows noninvasive and quantitative measurement of soft tissue motion and the changes in motion due to pathologic conditions. In cases in which the diagnosis of a tumor in the apex is in doubt, it may reduce the need for biopsy. In contrast to static computed tomographic (CT) scans and MRIs, it can differentiate between juxtaposition and continuity and may be a new and promising tool in the differential diagnosis of intraorbital lesions. PMID- 11006212 TI - Expression regulation of hyaluronan synthase in corneal endothelial cells. AB - PURPOSE: Our previous study showed that hyaluronan synthase (HAS), the enzyme protein of hyaluronan (HA) biosynthesis, is expressed in ocular tissues including the corneal endothelium. In the current study, the mechanism that regulates HAS expression in bovine corneal endothelial cells (BCECs) was investigated. METHODS: Cultured BCECs were used. HAS expression in BCECs at the mRNA level was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis. The effects of transforming growth factor (TGF)-beta and platelet derived growth factor (PDGF)-BB on HAS expression were examined by quantitative RT-PCR. The involvement of the Smad family (intracellular signal transducer of TGF-beta) was also investigated. The expression of HAS in BCECs at the protein level was confirmed by immunocytochemistry and Western blot analysis. RESULTS: Three HAS isoforms in BCECs were expressed at the mRNA level. The transcriptional sizes of each HAS in BCECs were 4.9 kb for HAS1, 2.8 kb for HAS2, and 1.6 kb for HAS3. The expression of HAS2 at the mRNA level was stimulated by TGF-beta1 and/or PDGF-BB treatment. In contrast, HAS1 and HAS3 expression was not affected by these growth factors. The additive effects of TGF-beta1 and PDGF-BB were observed in the stimulation of the expression levels of HAS2. HAS2 upregulation by these growth factors was also detected by Western blot analysis. The stimulation of the expression of HAS2 at the mRNA level by TGF-beta was accelerated by the overexpression of Smad2, Smad3, and Smad4 and inhibited by that of Smad7, all of which were confirmed to be involved in the signal transduction from TGF-beta through HAS expression. CONCLUSIONS: Although three HAS isoforms were expressed in the corneal endothelial cells, the expression of HAS2 was upregulated by TGF beta1 and/or PDGF-BB. HAS2 expression was regulated by TGF-beta through Smad family members. PMID- 11006213 TI - HRG4 (UNC119) mutation found in cone-rod dystrophy causes retinal degeneration in a transgenic model. AB - PURPOSE: To investigate the function and pathogenicity of HRG4, a photoreceptor synaptic protein homologous to the Caenorhabditis elegans neuroprotein UNC119. METHODS: HRG4 was screened for mutations in patients with various retinopathies, and a transgenic mouse model was constructed and analyzed based on a mutation found. RESULTS: A heterozygous premature termination codon mutation was found in a 57-year-old woman with late-onset cone-rod dystrophy. In some transgenic mice carrying the identical mutation, age-dependent fundus lesions developed accompanied by electroretinographic changes consistent with defects in photoreceptor synaptic transmission (depressed b-wave, normal c-wave), and retinal degeneration occurred with marked synaptic and possible transsynaptic degeneration. CONCLUSIONS: HRG4, the only synaptic protein known to be highly enriched in photoreceptor ribbon synapses, is now shown to be pathogenic when mutated. PMID- 11006214 TI - A new betaA1-crystallin splice junction mutation in autosomal dominant cataract. AB - PURPOSE: To map the locus for autosomal dominant cataracts (ADCs) in a Brazilian family using candidate gene linkage analyses, describe the clinical variability, and identify potential mutations in the human betaA1-crystallin gene (CRYBA1), a candidate gene identified through linkage studies demonstrating cosegregation with markers on chromosome 17. METHODS: Members of a Brazilian family with ADC were studied. Clinical examinations and linkage analyses with polymerase chain reaction (PCR) polymorphisms of 22 anonymous markers and 2 within the neurofibromatosis type 1 gene were performed; two-point lod scores were calculated. DNA sequences of all 6 exons and 12 exon-intron boundaries of the betaA1-crystallin gene, a proximal candidate gene mapped to 17q11.1-q12 in one unaffected and two affected individuals, were screened and new variants assessed for cosegregation with the disease. RESULTS: Affected individuals exhibited variable expressivity of pulverulent opacities in the embryonal nucleus and sutures; star-shaped, shieldlike, or radial opacities in the posterior embryonal nucleus; and/or midcortical opacities. All known loci for ADC in this family on chromosomes 1 and 13 were excluded. A positive lod score on chromosome 17 was calculated. This ADC locus was mapped to two potential regions on the long arm with an intervening recombination. The only known candidate gene in these regions was betaA1-crystallin. Three previously unreported single nucleotide variants were found in this gene, one in the donor splice junction site of intron C. This variant was found in all affected members and is presumed to be the causative mutation. CONCLUSIONS: An ADC locus was mapped in a Brazilian family with variable expressivity to either 17q23.1-23.2 or 17q11.1-12 based on linkage analyses. Analyses of DNA sequences of the betaA1-crystallin gene in this family revealed three new variants, one of which is within a donor splice junction and cosegregates with affected members. PMID- 11006215 TI - Ultrastructural and molecular analysis of Bowman's layer corneal dystrophies: an epithelial origin? AB - PURPOSE: Two mutations (R555Q and R124L) in the BIGH3 gene have been described in anterior or Bowman's layer dystrophies (CDB). The clinical, molecular, and ultrastructural findings of five families with CDB was reviewed to determine whether there is a consistent genotype:phenotype correlation. METHODS: Keratoplasty tissue from each patient was examined by light and electron microscopy (LM and EM). DNA was obtained, and exons 4 and 12 of BIGH3 were analyzed by polymerase chain reaction and single-stranded conformation polymorphism/heteroduplex analysis. Abnormally migrating products were analyzed by direct sequencing. RESULTS: In two families with type I CDB (CDBI), the R124L mutation was defined. There were light and ultrastructural features of superficial granular dystrophy and atypical banding of the "rod-shaped bodies" ultrastructurally. Patients from three families with "honeycomb" dystrophy were found to carry the R555Q mutation and had characteristic features of Bowman's dystrophy type II (CDBII). CONCLUSIONS: There is a strong genotype:phenotype correlation among CBDI (R124L) and CDBII (R555Q). LM and EM findings suggest that epithelial abnormalities may underlie the pathology of both conditions. The findings clarify the confusion over classification of the Bowman's layer dystrophies. PMID- 11006216 TI - Ascorbic acid concentration and total antioxidant activity of human tear fluid measured using the FRASC assay. AB - PURPOSE: To evaluate a novel method (FRASC) for total ferric reducing (antioxidant) activity and ascorbic acid concentration applied to human tears, to investigate the stability of ascorbic acid, and to determine the antioxidant status of human reflex tears. METHODS: Linearity, sensitivity, and precision of FRASC and ascorbic acid loss during 7 days' storage were assessed; total antioxidant activity and ascorbic acid and uric acid concentrations of reflex tears from 47 healthy subjects were measured. RESULTS: FRASC has good precision, linearity, and sensitivity. Ascorbic acid is stable for at least 7 days at moderately acidic pH (pH 3.6) and low temperature. Total antioxidant activity and ascorbic acid and uric acid concentrations (mean +/- SD) in reflex tears were 409 +/- 162, 23 +/- 9.6, and 68 +/- 46 microM, respectively. Ascorbic acid and uric acid constituted around half the total antioxidant activity measured. There was a significant correlation between uric acid and total antioxidant activity (r = 0.754; P: < 0.0001). Men had significantly (P: = 0.0045) higher tear ascorbic acid concentrations than women. CONCLUSIONS: FRASC is suitable for measuring total antioxidant activity and ascorbic acid in human tears. Further clinical study is needed to investigate the male-female difference seen, to characterize the remaining 50% antioxidant activity, and to investigate the effects of environmental conditions, antioxidant supplementation, age, and ocular disease on tear antioxidant status. PMID- 11006217 TI - Elevated levels of proteolytic enzymes in the aging human vitreous. AB - PURPOSE: To identify whether aging of human vitreous is accompanied by an elevation in degradative enzymes within the tissue. METHODS: Human vitreous samples from donors aged 10 to 88 years were placed in two groups based on donor age of less than or more than 50 years. Homogenized samples were analyzed by gelatin substrate zymography for matrix metalloproteinases (MMP). Serine proteinases were detected by casein substrate zymography, and a specific antibody was used to confirm the identity of, and to quantify, the serine proteinase, plasmin. RESULTS: Progelatinase A (ProMMP-2) was present in all the vitreous samples but did not show an age-related increase. MMP-2 was also present at low levels. Progelatinase B (ProMMP-9) was found in approximately 80% of samples analyzed, but neither its presence nor level of activity was age dependent. Of the serine proteinase activities detected, an enzyme of approximately 80 kDa was identified by Western blot analysis as plasmin(ogen). Quantitative analysis revealed a significant increase in plasmin(ogen) with age. CONCLUSIONS: This study shows there is an age-related increase in potential degradative activity in human vitreous that may be responsible for degenerative changes such as vitreous liquefaction. The data suggest that increased levels of these enzymes precede, or are indicative of, underlying ocular disease in some individuals. PMID- 11006218 TI - A novel resource for the study of genes expressed in the adult human retina. AB - PURPOSE: To reconstruct the transcriptional profile of the human adult retina and the genomic map of the genes expressed in this tissue. METHODS: Original software was used for the retrieval and analysis of records from UniGene (http://www.ncbi.nlm.nih. gov/UniGene/) pertaining to selected cDNA libraries from adult human retina. RESULTS: The 4974 genes reported so far to be expressed in retina were included in a catalog available on the Internet. For each entry, an estimation of the level of expression of the corresponding gene in the retina was provided. A high-resolution genomic map of the human retina was built up by inclusion of 3152 genes showing a precise and unique map assignment. The correspondence was established between 53 gene-orphan retinal diseases and clusters of genes expressed in the retina. CONCLUSIONS: The in silico reconstruction of the transcriptional profile of the adult human retina provides preliminary information on the pattern of genomic expression in this tissue. The chromosomal location of many retinal genes, combined with their expression data, should speed up the identification of genes involved in retinal diseases. PMID- 11006219 TI - Changing views on open-angle glaucoma: definitions and prevalences--The Rotterdam Study. AB - PURPOSE: To create a quantitative basis for diagnostic criteria for open-angle glaucoma (OAG), to propose an epidemiologic definition for OAG based on these, and to determine the prevalence of OAG in a general white population. METHODS: Of the 7983 subjects 55 years of age or older participating in the population-based Rotterdam Study, 6756 subjects participated in the ophthalmic part of this study (6281 subjects living independently and 475 in nursing homes). The criteria for the diagnosis of OAG were based on ophthalmoscopic and semiautomated Imagenet estimations of the optic disc such as vertical cup-to-disc ratio (VCDR), minimal width of neural rim, or asymmetry in VCDR between both eyes, and visual field testing with kinetic Goldmann perimetry. All criteria for the diagnosis of OAG were assessed in a masked way independently of each other. RESULTS: Mean VCDR on ophthalmoscopy was 0.3 and with Imagenet 0.49, and the 97.5th percentile for both was 0.7. The prevalence of glaucomatous visual field defects was 1.5%. Overall prevalence of definite OAG in the independently living subjects was 0.8% (95% confidence interval [CI] 0.6, 1.0; 50 cases). Prevalence of OAG in men was double that in women (odds ratio 2.1; 95% CI 1.2, 3.6). Different commonly used criteria for diagnosis of OAG resulted in prevalence figures ranging from 0.1% to 1.2%. CONCLUSIONS: The overall prevalence of OAG in the present study was comparable to most population-based studies. However, prevalence figures differed by a factor of 12 when their criteria for OAG were applied to this population. A definition for definite OAG is proposed: a glaucomatous optic neuropathy in eyes with open angles in the absence of history or signs of secondary glaucoma characterized by glaucomatous changes based on the 97.5 percentile for this population together with glaucomatous visual field loss. In the absence of the latter or of a visual field test, it is proposed to speak of probable OAG based on the 99.5th or possible OAG based on the 97.5th percentiles of glaucomatous disc changes for a population under study. PMID- 11006220 TI - Influence of lutein supplementation on macular pigment, assessed with two objective techniques. AB - PURPOSE: Macular pigment (MP) may protect against age-related macular degeneration. This study was conducted to determine the extent of changes in the macular pigment density as a consequence of oral supplementation with lutein. A second purpose was to compare two objective measurement techniques. METHODS: In the first technique, reflectance maps were made with a scanning laser ophthalmoscope. Digital subtraction of log reflectance maps and comparison between the foveal area and a 14 degrees temporal site provided MP density estimates. In the second technique, spectral fundus reflectance of the fovea was measured with a fundus reflectometer and analyzed with a detailed optical model, to arrive at MP density values. Eight subjects participated in this study. They took 10 mg lutein per day for 12 weeks. Plasma lutein concentration was measured at 4-week intervals. RESULTS: After 4 weeks, mean blood level of lutein had increased from 0.18 to 0.90 microM. It stayed at this level throughout the intake period and declined to 0.28 microM 4 weeks after termination. Measurement of the density of MP showed a within-subject variation of 10% with MP maps and 17% with spectral reflectance analysis. MP density showed a mean linear 4-week increase of 5.3% (P: < 0.001) and 4.1% (P: = 0. 022), respectively. CONCLUSIONS: Supplementation with lutein significantly increased the density of the MP. Analyzing reflectance maps with a scanning laser ophthalmoscope provided very reliable estimates of MP. PMID- 11006221 TI - Monocular versus binocular visual acuity as measures of vision impairment and predictors of visual disability. AB - PURPOSE: To examine the relationship between monocular and binocular visual acuities as predictors of visual disability in a population-based sample of individuals 65 years of age and older. METHODS: Two thousand five hundred twenty community-dwelling residents of Salisbury, Maryland, between the ages of 65 and 84 years of age were recruited for the study. Corrected visual acuity was measured monocularly and binocularly using ETDRS charts. Reading speed, face discrimination, and self-reported difficulty with visual tasks were also determined. RESULTS: Binocular acuity is predicted with reasonable accuracy by acuity in the better eye alone, but not by the widely used American Medical Association (AMA) weighted-average algorithm. The AMA algorithm significantly underestimates binocular acuity when the interocular acuity difference exceeds one line. Monocular acuity and binocular acuity were significantly better predictors of reading speed than the AMA weighted score or a recently proposed Functional Vision Score (FVS). Monocular acuity in the better eye, binocular acuity, and the AMA and FVS algorithms were equally good predictors of self reported vision disability. None of the acuity measures were good predictors of face recognition ability. CONCLUSIONS: The binocular acuities of older individuals can be inferred from measures of monocular acuity. There is little evidence for binocular inhibition when the monocular acuities in the two eyes are unequal, as opposed to the widely used AMA algorithm for computing binocular visual impairment. For tasks that are strongly associated with visual acuity, such as reading, this association can be captured from measures of monocular acuity and does not require separate assessment of binocular acuity. PMID- 11006222 TI - Photochemical keratodesmos for repair of lamellar corneal incisions. AB - PURPOSE: To determine the efficacy of photochemical keratodesmos (PKD) for closing surgical incisions in the cornea of enucleated rabbit eyes compared with that achieved using sutures and self-sealing incisions. METHODS: A 3.5-mm incision, at an angle parallel to the iris, was made in the cornea of enucleated New Zealand White rabbit eyes. The intraocular pressure required to cause leakage (IOP(L)) from the untreated incision was then recorded. Photochemical keratodesmos treatment was then performed by application of a dye, Rose Bengal (RB), in saline solution to the surfaces of the incision wound, followed by laser irradiation at 514 nm from an argon ion laser. Immediately after treatment, the IOP(L) was measured. Both dose and laser irradiance dependencies were studied in five or more eyes for each condition and appropriate control eyes. The IOP(L)s were compared with those obtained using conventional interrupted 10-0 nylon sutures. Other dyes were tested in a similar fashion. RESULTS: The IOP(L) of 300 mm Hg was obtained using a fluence of 1270 J/cm(2) with an irradiance of 1.27 W/cm(2) (laser exposure time, 16 minutes 40 seconds). No sealing was observed using dye or light alone where control pressures of approximately 30 mm Hg were found. At higher dose (1524 J/cm(2)) and irradiance (3.82 W/cm(2); 6 minutes 35 seconds), PKD was less effective, which may be attributable to thermal effects. PKD produced IOP(L)s similar to those in closure by sutures. Other dyes such as riboflavin-5-phosphate and N:-hydroxy-pyridine thione also produced efficient bonding after PKD. Nonphotochemically active dyes did not produce significant increases in the IOP(L) at which leakage occurred. CONCLUSIONS: The increase in IOP(L) after PKD treatment, comparable with that with sutures, in enucleated rabbit eyes demonstrates the feasibility of this technique ex vivo. PMID- 11006223 TI - The role of cytotoxic T lymphocytes in corneal allograft rejection. AB - PURPOSE: Immunologic rejection constitutes a major barrier to the success of allogeneic corneal transplants, but the specific mediators and mechanisms of graft rejection are poorly understood. Several studies have implicated cytotoxic T-lymphocyte (CTL) responses, typically associated with CD8(+) T cells, in promoting corneal graft rejection. This study sought to test the hypothesis that CTLs are essential in promoting corneal graft rejection. METHODS: BALB/c donor corneas were grafted orthotopically onto C57BL/6, perforin knockout, or CD8(+) T cell knockout mice. The tempo and incidence of graft rejection were observed for each group. In separate experiments, donor-specific CTL and delayed-type hypersensitivity (DTH) responses were tested at the time of graft rejection by a standard chromium release assay and an ear swelling assay, respectively. RESULTS: Perforin knockout and CD8(+) T-cell knockout mice were as effective as wild-type C57BL/6 control mice in rejecting BALB/c donor corneas. Furthermore, animals in all three groups were found to develop robust donor-specific DTH, not CTL, responses at the time of graft rejection. Histopathologically, the rejected corneas from all three groups contained a predominantly mononuclear cellular infiltrate. CONCLUSIONS: This study rejects the hypothesis that CD8(+) CTLs are essential in promoting corneal graft rejection and instead further implicates donor-specific DTH reactions as the relevant immune response during graft failure. PMID- 11006224 TI - The thickness of the human precorneal tear film: evidence from reflection spectra. AB - PURPOSE: Interferometric methods have considerable potential for studying the thickness of layers of the human tear film and cornea because of their ability to make noninvasive, accurate, and rapid measurements. However, previous interferometric studies by Prydal and Danjo yielded tear thickness values near 40 and 11 microm, respectively, considerably greater than estimates made by invasive methods of 4 to 8 microm. Using a modified version of Danjo's method, interference effects from the tear film and cornea were studied, with the aim of correlation with known structure and optical properties of the cornea and hence determining the most probable value of tear film thickness. METHODS: Reflectance spectra from the human cornea were measured at normal incidence. These spectra show oscillations whose maxima correspond to constructive interference between light reflected from the air surface and from some deeper surface. The frequency of these spectral oscillations is proportional to the thickness of the layer between the air surface and the second surface. Therefore, Fourier analysis of reflectance spectra can be used to determine the thickness of layers of the tear film and cornea. In the main experiment, 36 low-resolution spectra were obtained from six normal eyes for measuring thickness up to 100 microm. Control experiments included measurements of the time course of thickness changes and high-resolution spectra for measuring thickness up to 1000 microm. RESULTS: For the main experiment, in the thickness range 1 to 100 microm, the strongest peak in the Fourier transform was near 3 microm (range, 1.5-4.7 microm) beneath the air surface. In the range 20 to 100 microm, the strongest peak was near 55 microm (range, 50-59 microm) for all 36 spectra; none were in Prydal's range near 40 microm. This 55-microm peak is consistent with a reflection from the basement membrane of the epithelium. Time course measurements after a blink show that the 3-microm peak is not an artifact. High-resolution spectra gave a peak near 510 microm, corresponding to the complete thickness of the cornea (plus tear film). This peak had a contrast similar to that of the 3-microm peak. CONCLUSIONS: These studies did not confirm Prydal's estimate of approximately 40 microm. Nor were there prominent peaks near Danjo's value of approximately 11 microm, except in cases of probable reflex tears. Because the reflection at the aqueous-mucus boundary would be expected to be weaker than that from the epithelial surface, the 3-microm peak is unlikely to correspond to the aqueous layer (rather than the complete tear film). The proposal that the 3-microm peak corresponds to a reflection from the front of the cornea is supported by the demonstration of a peak of similar contrast from the back of the cornea. Thus, the current evidence consistently supports a value of approximately 3 microm for the thickness of the human precorneal tear film. PMID- 11006225 TI - Apoptosis in shed human corneal cells. AB - PURPOSE: To determine whether shear forces applied to the corneal epithelium by the repeated insertion and removal of a hydrogel contact lens alter the size and number of cells removed and to determine the contribution of apoptosis to this process. METHODS; Human corneal cells were collected from eight healthy subjects by sequential contact lens cytology (20 lens insertions and removals). Collected cells were stained with acridine orange for counting and measurement of cell size. In a separate experiment, collected cells were fixed and stained with TdT mediated dUTP nick-end labeling (TUNEL) or labeled immediately after collection using annexin V. Hoechst stain and propidium iodide (PI) were used as nuclear counterstains. The proportion of cells labeled with acridine orange, TUNEL, and annexin V was quantified by fluorescence microscopy. RESULTS; The number of cells increased in later collections, and cells were smaller. The mean number of positively stained cells using TUNEL was 57%. Annexin V labeling on unfixed fresh samples showed a mean of 64%, with an increase in later collections. Apoptotic bodies were observed in very few cells. In most cells the nucleus and cytoplasmic membrane were intact. Structures were observed in which nuclei were missing (Hoechst negative) but in which cytoplasm had the size and appearance of whole, nucleated cells. These structures (cell ghosts) increased in number along with the increase in nucleated cells in later collections. CONCLUSIONS: The sequential removal of a soft contact lens caused a progressive increase in the number of cells collected from the surface and a progressive decrease in their size. The majority of nucleated cells removed by a contact lens were apoptotic in the sense of being positively labeled by TUNEL and annexin V. Morphologically they differed from classically apoptotic cells, in that cells showed an intact nuclear structure and no discernible apoptotic bodies. They could represent a last stage in a pathway of cell differentiation in which frictional forces induced by the removal of the contact lens activate the apoptotic program and cause the cell to be shed. There is also a pathway in which cells lose their nuclei before leaving the epithelial surface. PMID- 11006226 TI - Corneal opacity in lumican-null mice: defects in collagen fibril structure and packing in the posterior stroma. AB - PURPOSE: Gene targeted lumican-null mutants (lum(tm1sc)/lum(tm1sc)) have cloudy corneas with abnormally thick collagen fibrils. The purpose of the present study was to analyze the loss of transparency quantitatively and to define the associated corneal collagen fibril and stromal defects. METHODS: Backscattering of light, a function of corneal haze and opacification, was determined regionally using in vivo confocal microscopy in lumican-deficient and wild-type control mice. Fibril organization and structure were analyzed using transmission electron microscopy. Biochemical approaches were used to quantify glycosaminoglycan contents. Lumican distribution in the cornea was elucidated immunohistochemically. RESULTS; Compared with control stromas, lumican-deficient stromas displayed a threefold increase in backscattered light with maximal increase confined to the posterior stroma. Confocal microscopy through-focusing (CMTF) measurement profiles also indicated a 40% reduction in stromal thickness in the lumican-null mice. Transmission electron microscopy indicated significant collagen fibril abnormalities in the posterior stroma, with the anterior stroma remaining relatively unremarkable. The lumican-deficient posterior stroma displayed a pronounced increase in fibril diameter, large fibril aggregates, altered fibril packing, and poor lamellar organization. Immunostaining of wild type corneas demonstrated high concentrations of lumican in the posterior stroma. Biochemical assessment of keratan sulfate (KS) content of whole eyes revealed a 25% reduction in KS content in the lumican-deficient mice. CONCLUSIONS: The structural defects and maximum backscattering of light clearly localized to the posterior stroma of lumican-deficient mice. In normal mice, an enrichment of lumican was observed in the posterior stroma compared with that in the anterior stroma. Taken together, these observations indicate a key role for lumican in the posterior stroma in maintaining normal fibril architecture, most likely by regulating fibril assembly and maintaining optimal KS content required for transparency. PMID- 11006227 TI - Erk and PI-3 kinase are necessary for collagen binding and actin reorganization in corneal epithelia. AB - PURPOSE: It was recently shown that phosphatidylinositol-(PI)3 kinase is upregulated in wounded rabbit corneal epithelia. Extracellular signal-regulated kinase (erk)-1 and -2 proteins and PI-3 kinase were activated in embryonic corneal epithelia after 1-hour stimulation by type I collagen. In the current investigation specific inhibitors of PI-3 kinase and mitogen-activated kinase kinase (MEK-1 kinase) were used to determine the role of these signaling molecules in actin reorganization and collagen binding to isolated sheets of corneal epithelial tissue. METHODS: Effects of specific PI-3 kinase and MEK-1 inhibitors (LY294002, PD98059, respectively) were investigated in embryonic corneal epithelial tissues. Avian embryonic corneal epithelia were isolated as tissue sheets, organ cultured in the presence of these specific inhibitors, and stimulated with type I collagen. The tissues were evaluated for collagen stimulated actin reorganization, erk-1 and -2 and PI-3 kinase activity, total filamentous actin accumulation, and collagen binding. RESULTS: The MEK-1 inhibitor PD98059 decreased erk-1 and -2 phosphorylation and blocked actin reorganization in a dose-dependent manner. The PI-3 kinase 85-kDa subunit was decreased 25% in LY294002-treated tissue, and collagen binding also decreased significantly in tissues treated with MEK-1 and PI-3 kinase inhibitors compared with control tissues. In addition, both inhibitors blocked actin cortical mat reorganization. CONCLUSIONS; PI-3 kinase and erk-1 and -2 signaling pathways are activated and necessary for collagen binding and integrin-mediated actin reorganization in embryonic avian corneal epithelium. PMID- 11006228 TI - Localization of thrombomodulin in the anterior segment of the human eye. AB - PURPOSE: To localize thrombomodulin (TM) in the anterior segment of the human eye. TM is a vascular endothelial cell surface glycoprotein that acts as a cofactor for the thrombin-catalyzed activation of the anticoagulant protease zymogen, protein C. METHODS: Immunohistochemical methods were used to detect TM expression in corneal epithelial cells, the lens epithelial cells, and other cells in the anterior segment of the eye. The expression of TM was also examined in cultured human corneal epithelial cells. RESULTS: TM was expressed in corneal epithelial cells, corneal endothelial cells, and nonpigmented ciliary epithelial cells, which are in direct contact with the aqueous humor. TM was also expressed in cultured corneal epithelial cells and showed cofactor activity. The amount of the antigen in the cultured corneal cells was approximately one tenth of that in human umbilical vein endothelial cells, but its specific cofactor activity (75%) was comparable to that of TM in human umbilical vein endothelial cells. The trabecular meshwork and endothelial cells lining Schlemm's canal also showed positive staining for TM. CONCLUSIONS: The TM in the cells that are in contact with the aqueous humor appears to be involved in maintaining the fluidity of the aqueous humor. In contrast, TM in cells that are not in contact with the aqueous humor may function in regulating cell proliferation and/or differentiation. PMID- 11006229 TI - The distribution of myosin heavy chain isoforms among rat extraocular muscle fiber types. AB - PURPOSE: To determine the distribution of myosin heavy chain isoforms in each extraocular muscle (EOM) fiber type. METHODS: Serial sections of adult rat EOMs were stained with isoform-specific monoclonal antibodies against an array of myosin heavy chains. Immunofluorescent antibody staining of whole adult rat EOMs, examined by confocal microscopy, demonstrated the longitudinal variations of isoforms along individual fibers. RESULTS: Each global fiber type reacted predominantly with a single isoform-specific antibody and showed no longitudinal variation. Two major orbital fibers were defined, and both contained multiple myosin heavy chains. Both orbital singly and multiply innervated fibers stained proximal and distal to the neuromuscular junction with antibody to embryonic myosin heavy chain, but this isoform was sharply and completely excluded from the domain of the neuromuscular junction. Orbital singly innervated fibers also contained the EOM-specific isoform at the neuromuscular junction. Orbital multiply innervated fibers did not contain the EOM-specific isoform, but additionally contained a slow isoform along their entire length. CONCLUSIONS: Adult rat EOMs show unique fiber types with arrangements of myosin heavy chain isoforms not seen in other skeletal muscles. Moreover, unique cellular mechanisms must exist to target each isoform to its proper domain along individual orbital fibers. PMID- 11006230 TI - Histologic effect of mitomycin C on strabismus surgery in the rabbit. AB - PURPOSE: To evaluate the efficiency of mitomycin C (MMC) in limiting the postoperative inflammatory response and scarring after strabismus surgery. METHODS: A prospective, two-stage, masked, controlled trial was conducted. In the first stage, the inflammatory response at the extraocular muscle reattachment site was increased after inferior rectus recession in eight rabbits. In the second stage, MMC (0.4 mg/ml) was applied during surgery to the eyes of 22 rabbits with inferior rectus recession. As a control, contralateral eyes were treated with saline solution. Seven weeks later, exenteration was performed, and the sites of muscle reattachment were processed for histologic examinations. The sums of the areas of the granulomas in the extraocular muscle reattachment sites of control and treated eyes were compared. RESULTS: There was no significant inhibitory effect of MMC on the inflammatory response of treated eyes compared with that of control eyes. CONCLUSIONS: The intraoperative use of MMC (0.4 mg/ml) was not effective in controlling the postoperative inflammatory response in rabbit eyes after extraocular muscle surgery. These data do not support the hypothesis that MMC reduces postoperative adhesions after strabismus surgery. PMID- 11006231 TI - Acute effects of the skeletal muscle-specific immunotoxin ricin-mAb 35 on extraocular muscles of rabbits. AB - PURPOSE: To determine the acute histologic and ultrastructural effects of a recently developed muscle-specific immunotoxin, ricin-mAb 35. METHODS: Graduated doses of ricin-mAb 35, composed of ricin conjugated to a monoclonal antibody against the nicotinic acetylcholine receptor of skeletal muscle, were injected into one superior rectus muscle in rabbits. After 3, 7, and 14 days, both superior rectus muscles were removed and prepared for electron microscopy and histologic examination, by using a number of immunohistochemical markers to identify inflammatory cell infiltration, muscle fiber loss, and muscle regeneration. RESULTS: Myotoxicity of the ricin-mAb 35 was focal and dose related. At the highest dose tested, there was substantial inflammatory cell infiltrate by 3 days, which largely disappeared by 7 days. Significant muscle loss was apparent by 7 days after ricin-mAb 35 treatment. Both the inflammatory reaction and muscle fiber loss were confined to the immediate injection site. Surrounding muscle appeared to be normal. At 14 days after treatment, early signs of muscle regeneration were evident within the tissue sections. No evidence of orbital or systemic toxicity was seen in any animal. CONCLUSIONS: Direct injection of ricin-mAb 35 into the extraocular muscles of rabbits results in a dose-related focal injury to the muscles, with a self-limited inflammatory component and significant muscle fiber loss. This novel immunotoxin may be useful in the treatment of strabismus if chronic studies show a sustained histologic and electrophysiologic effect. PMID- 11006232 TI - Infant hyperacuity for radial deformation. AB - PURPOSE: Poor response rates and/or the confounding of motion and offset responses make it difficult to interpret results of previous studies of infant hyperacuity. The aim of the present study was to design a protocol that overcomes these limitations and to investigate the normal maturation of hyperacuity. METHODS: Hyperacuity of 31 healthy term infants aged 4 to 12 months was measured using radial deformation of static circular D4 patterns with a two-alternative, forced-choice, preferential-looking (FPL) protocol and maximum likelihood threshold estimation. FPL grating resolution acuity was assessed on the same visit. RESULTS: Both hyperacuity and resolution acuity were 1.1 to 1.2 logMAR (12 16 minutes arc) at 4 months of age. Hyperacuity improved rapidly to approximately 0.3 logMAR (2.0 minutes arc) by 9 to 12 months of age. This 0.9 log unit improvement in the hyperacuity still leaves the 12-month-old infant at a level 0.4 log unit poorer than adults' thresholds. Resolution acuity improved more gradually to approximately 0.7 logMAR (5 minutes arc) by 9 to 12 months of age. This 0.4 log unit improvement leaves the 12-month-old infant at a level 0.6 log unit poorer than adults' resolution acuity. CONCLUSIONS: Hyperacuity measured via radial deformation thresholds matures very rapidly between 4 and 6 months of age and continues to mature more slowly throughout infancy and into early childhood. The radial deformation protocol may provide a sensitive index for detecting and monitoring abnormalities in spatial vision in cases of infantile esotropia. PMID- 11006233 TI - Effect of general anesthetics on IOP in rats with experimental aqueous outflow obstruction. AB - PURPOSE: To determine the effect of several common general anesthetics on intraocular pressure (IOP) after experimental aqueous outflow obstruction in the rat. METHODS: A single episcleral vein injection of hypertonic saline was used to sclerose aqueous humor outflow pathways and produce elevated IOP in Brown Norway rats. Animals were housed in either standard lighting or a constant low-level light environment. Awake IOPs were determined using a TonoPen (Mentor, Norwell, MA) immediately before induction of anesthesia by either isoflurane, ketamine, or a mixture of injectable anesthetics (xylazine, ketamine, and acepromazine). For each anesthetic, IOPs were measured immediately after adequate sedation (time 0) and at 5-minute intervals, up to 20 minutes. RESULTS; Awake IOPs ranged from 18 to 52 mm Hg. All anesthetics resulted in a statistically significant (P: < 0.01) reduction in measured IOP at every duration of anesthesia when compared with the corresponding awake IOP. With increasing duration of anesthesia, measured IOP decreased approximately linearly for both the anesthetic mixture and isoflurane. However, with ketamine, IOP declined to 48% +/- 11% (standard lighting) and 60% +/- 7% (constant light) of awake levels at 5 minutes of anesthesia, where it remained stable. In fellow eyes, the SD of the mean IOP in animals under anesthesia was always greater than the corresponding SD of the awake mean. Anesthesia's effects in normal eyes and eyes with elevated IOP were indistinguishable. CONCLUSIONS: All anesthetics resulted in rapid and substantial decreases in IOP in all eyes and increased the interanimal variability in IOPs. Measurement of IOP in awake animals provides the most accurate documentation of pressure histories for rat glaucoma model studies. PMID- 11006234 TI - Localization of MYOC transcripts in human eye and optic nerve by in situ hybridization. AB - PURPOSE: To evaluate MYOC (myocilin) gene expression at the RNA level in normal intact human eyes and optic nerve using in situ hybridization. METHODS: Normal human eyes and optic nerves from donors 62 to 83 years of age with no history of glaucoma were fixed, embedded in paraffin, and sectioned. Sections were hybridized with (35)S-labeled sense and antisense riboprobes derived from a full length MYOC cDNA. RESULTS: High levels of MYOC expression were observed throughout the trabecular meshwork as well as in the most anterior nonfiltering meshwork (Schwalbe's line), in the scleral spur, and in the endothelial lining of Schlemm's canal. MYOC transcripts were also detected in the anterior corneal stroma, in the ciliary muscle, beneath the anterior border of the iris, in the iris stroma, and in the sclera. Expression in the retrolaminar region of the optic nerve was present in the pial septa that divide the nerve fiber bundles, in the perivascular connective tissue surrounding the central retinal vessels, and in the dura mater, arachnoid, and pia mater of the meningeal sheath surrounding the optic nerve. CONCLUSIONS: MYOC gene expression in the trabecular meshwork, Schlemm's canal, scleral spur, and ciliary muscle indicates a structural or functional role for myocilin in the regulation of aqueous humor outflow that may influence intraocular pressure. MYOC expression in the optic nerve suggests that changes in the structural, metabolic, or neurotropic support of the optic nerve may influence its susceptibility to glaucomatous damage. PMID- 11006235 TI - The long-term fluctuation of the visual field in stable glaucoma. AB - PURPOSE: To determine, in stable glaucoma, the characteristics of the between examination variability of the visual field recorded with the Humphrey Field Analyser (HFA; Humphrey Systems, Dublin, CA) using the homogeneous, LF(Ho), and heterogeneous, LF(He), components of the long-term fluctuation (LF), thereby providing a technique for separating progressive loss from fluctuation in sensitivity. METHODS: The LF components were calculated using a two-factor analysis of variance (ANOVA) with replications and were determined between each pair of three successive HFA program 30-2 fields for each patient from two groups, each containing 30 patients with primary open-angle glaucoma. The interval between examinations for the first group was 6 to 9 months and for the second group was 3 weeks. RESULTS: The group mean values for LF(Ho) ranged from 1.50 to 2.19 dB and for LF(He) from 1.70 to 2.05 dB. The average difference between examinations was within +/-0.35 dB for each component, and the 95% limits of agreement for the two groups, respectively, were +/- 2.31 and +/- 2.39 dB for the LF(Ho) and +/- 2.36 and +/- 2.09 dB for the LF(He). The estimate of the 90% confidence limit for the LF(Ho) was 3.30 dB and for the LF(He), 3.60 dB. Little relationship was present between the LF components and the modulus differences in mean deviation (MD), the corrected pattern SD (CPSD), or the mean MD, mean short term fluctuation, and mean CPSD, of the two fields. CONCLUSIONS: Estimation of the LF components and of the corresponding confidence limits yields an expression of the normal between-examination variability of two consecutive fields that can be used as a reliability index. A value outside the confidence limits indicates the necessity for a confirmatory follow-up field. PMID- 11006236 TI - Modulation of pre-mRNA splicing and protein production of fibronectin by TGF beta2 in porcine trabecular cells. AB - PURPOSE: To determine the effect of transforming growth factor (TGF)-beta2 on the pre-mRNA splicing pattern of fibronectin, as well as on the synthesis and secretion of this glycoprotein by porcine trabecular cells. METHODS: First passage porcine trabecular cells were rendered quiescent and incubated in culture medium containing 15% newborn calf serum, in serum-free culture medium containing either activated TGF-beta2 (concentration range: 0.2-2.7 ng/ml) or activated TGF beta1 (1 ng/ml), or in serum-free medium alone (untreated control samples). For investigation of alternative splicing, total RNA was extracted, and reverse transcription-polymerase chain reaction (RT-PCR) was performed with primer pairs located in exons flanking the exon (extra domain [ED]A, or EDB) that undergoes alternative splicing. The polymerase chain reaction (PCR) products were verified by Southern hybridization and quantified by using laser densitometry. The percentage of EDA-positive (+) isoforms was compared with that of the EDB+ isoforms among the groups. To study the effect of TGF-beta2 on the synthesis and secretion of fibronectin, total protein was extracted from both cultured cells and conditioned medium, Western blot analysis was performed using an anti fibronectin antibody, and the products were quantified by laser densitometry. Immunocytochemical analysis was also performed on cultured trabecular cells to detect fibronectin. RESULTS: Fibronectin mRNA that was detected in untreated serum-starved control cells was EDA and EDB negative. Incubation of trabecular cells in medium containing 1 ng/ml TGF-beta2, 1 ng/ml TGF-beta1, or 15% newborn calf serum induced the expression of EDA+ and EDB+ mRNA to varying degrees. At concentrations of 0.2, 0.5, 1.5, and 2.7 ng/ml, TGF-beta2 increased the concentration of fibronectin by 2-, 3-, 3.8-, and 5-fold in the conditioned medium, and by 3-, 3.7-, 4-, and 4.3-fold in the cell extracts, respectively. The trabecular cells treated with TGF-beta2 exhibited strong immunoreaction for fibronectin, whereas the cells incubated in serum-free medium showed only minimal immunoreactivity. CONCLUSIONS: Our results demonstrate that TGF-beta2 and TGF beta1 modified the alternative splicing pattern of fibronectin pre-mRNA and enhanced the synthesis and secretion of this extracellular matrix molecule by trabecular cells in a dose-dependent fashion. These findings indicate a mechanism whereby TGF-beta2, the concentration of which is elevated in aqueous humor of patients with primary open-angle glaucoma, contributes to the increased deposition of extracellular matrix molecules in the outflow pathway. PMID- 11006237 TI - Muller cell protection of rat retinal ganglion cells from glutamate and nitric oxide neurotoxicity. AB - PURPOSE: Low concentrations of excitotoxic agents such as glutamate and nitric oxide decrease survival rates of purified retinal ganglion cells (RGCs). In the retina, RGCs are ensheathed by retinal Muller glial (RMG) cell processes. The purpose of this study was to determine whether RMG cells could protect RGCs from these excitotoxic injuries. METHODS: RGCs were purified from 7- or 8-day-old Long Evans rats and cultured on polylysine/laminin-coated coverslips in serum-free medium for 2 days. The coverslips were then moved to dishes containing either confluent RMG monolayers or no glial cells in glutamate-free medium. Some dishes with confluent RMG cells were exposed to D,L-threo-beta-hydroxyaspartate (THA), a blocker of glutamate uptake. Three days after exposure to various concentrations of glutamate or the NO donor, 2, 2'-(hydroxynitroso-hydrazino)bisethanamine, survival rates of RGCs were measured by calcein-acetoxymethyl ester staining. Glutamate concentrations in the medium were measured using amino acid analysis. RESULTS: Without RMG cells, the application of increasing concentrations (5-500 microM) of glutamate caused a dose-dependent increase in RGC death after 3 days. The neurotoxic effects of glutamate were blocked in the RMG cell cocultures, even when there was no direct contact between the cell types. The protective effect of RMG cells was weakened by THA treatment. NO also had toxic effects on RGC. RMG cells prevented this toxicity but only when in direct contact with the RGCs. CONCLUSIONS: RMG cells can protect RGCs from glutamate and NO neurotoxicity. We suggest that functional disorders of glutamate uptake in RMGs might be one of the etiologies of glaucoma. PMID- 11006238 TI - Retinal damage after 3 to 4 months of elevated intraocular pressure in a rat glaucoma model. AB - PURPOSE: To characterize a long-term elevated intraocular pressure (IOP) glaucoma model in the rat with respect to electroretinographic (ERG) changes and the pattern and mechanism of retinal ganglion cell (RGC) death. METHODS; An approximate doubling of IOP was induced in one eye (G) of female Wistar rats (150 180 g) by cautery of 3 episcleral/limbal veins. At intervals over 3 to 4 months, measurements of IOP and ERG changes (contact-lens electrode) were made in both the G and contralateral normal (N) eyes. At the end of 3 to 4 months of elevated IOP, RGCs were fluorescently labeled with Fluorogold (retrogradely from the superior colliculus), or retinas were labeled by intravitreal injection of a mitochondrial potential indicator dye and stained for apoptotic nuclei with a DNA dye. Flatmounts of fixed, dye-labeled retinas were examined by epifluorescence, confocal, or interference contrast microscopy. RESULTS: Elevated IOP was consistently maintained for up to 4 months in G eyes, but ERG a- and b-waves showed a statistically significant decline, of 30% to 40% in amplitude, after 3 months. Loss of RGCs in G retinas was primarily focal with no statistically significant loss demonstrable outside of the focal areas when assessed by an area sampling method for counting RGCs, which totaled 2% to 3% of the entire retinal area. Mitochondrial membrane potential of cells in the RGC layer was reduced by 17.5% (P: < 0.05) in regions surrounding areas of focal loss compared with comparable locations in control N eyes. After 3.5 months' elevated IOP the G retinas showed cell nuclei at various stages of apoptosis, from initial DNA condensation to fragmentation. CONCLUSIONS: The three-vein episcleral/limbal vein occlusion model for inducing glaucomatous pathology in the rat eye gives a consistent long-term elevation of IOP. After 3 to 4 months of approximately 100% increased IOP, the ERG responses begin to decline, there is a variable focal loss of RGCs, and some of the remaining RGCs show characteristics of stress and apoptosis. These changes seem consistent with retinal damage in human glaucoma (focal field defects), and this rat model appears to mimic some features of primary open-angle glaucoma. PMID- 11006239 TI - Retrograde axonal transport of BDNF in retinal ganglion cells is blocked by acute IOP elevation in rats. AB - PURPOSE: To determine whether acute experimental glaucoma in rats obstructs retrograde transport of brain-derived neurotrophic factor (BDNF) to retinal ganglion cells (RGCs). METHODS: Forty rats had unilateral injection of either (125)I-BDNF (20 animals) or a mixture of (125)I-BDNF and 100-fold excess nonradiolabeled BDNF (20 animals). In each group of 20 animals, eyes contralateral to injection had either normal intraocular pressure (IOP; 10 animals) or IOP elevated to 25 mm Hg below the systolic blood pressure of the eye (10 animals). In each group of 20 rats, ipsilateral eyes had IOP set at systolic blood pressure (4 eyes), had optic nerve transection (10 eyes), or had normal IOP (6 eyes). Six hours after injection, animals were killed and tissues were fixed, embedded, and sectioned for autoradiography. Grain counts were performed over retina and optic nerve using automated image analysis. RESULTS: IOP elevation to 25 mm Hg below systolic blood pressure (perfusion pressure [PP] 25) decreased median retinal nerve fiber layer (NFL) grains by 38% compared with controls (P: < 0.001). Competition by cold BDNF reduced NFL grains by 28% (P: = 0.013). Considering only the radioactivity representing specific retrograde transport of BDNF, IOP elevation to PP25 reduced transport by 74%, whereas elevation to PP0 (equaling systolic blood pressure) reduced specific transport by 83%. CONCLUSIONS: BDNF is transported retrogradely from the superior colliculus in adult rats, and this transport is substantially inhibited by acute IOP elevation. Deprivation of BDNF among RGCs may contribute to neuron loss in glaucoma. PMID- 11006240 TI - Evidence that retinal pigment epithelium functions as an immune-privileged tissue. AB - PURPOSE: Tissues derived from immune-privileged sites sometimes possess special characteristics that promote their own survival when transplanted to a nonprivileged site. This study was undertaken to evaluate whether retinal pigment epithelium (RPE) behaves as an immune-privileged tissue when transplanted extraocularly. METHODS: RPE grafts were prepared from eyes of neonatal C57BL/6 or C57BL/6 gld/gld (deficient in CD95 ligand expression) mice. These grafts (or conjunctival grafts as positive controls) were transplanted into the anterior chamber, the subretinal space, the subconjunctival space, and underneath the kidney capsule of histoincompatible BALB/c mice. Transplant survival was evaluated by histology at selected time points after engraftment. Recipients were tested for acquisition of C57BL/6-specific delayed-type hypersensitivity (DH) and for the ability to suppress DH. RESULTS: Allogeneic neonatal RPE grafts from normal donors showed significantly enhanced survival at all graft sites compared with conjunctival grafts. However, allogeneic RPE cell grafts from gld/gld mice were rapidly rejected after transplantation beneath the kidney capsule. Allogeneic RPE grafts placed in extraocular sites induced systemic DH directed at donor alloantigens, whereas RPE allografts placed intraocularly induced suppression of systemic DH. CONCLUSIONS: Allogeneic neonatal RPE grafts, through constitutive expression of CD95 ligand, promote their own survival at heterotopic sites. Paradoxically, these grafts also display immunogenicity. Thus, neonatal RPE tissue owes its immune privilege to the capacity to prevent immune rejection rather than to inhibit sensitization. PMID- 11006241 TI - PCR-based evidence of bacterial involvement in eyes with suspected intraocular infection. AB - PURPOSE: To assess the usefulness of polymerase chain reaction (PCR) in detection of bacteria in ocular samples. METHODS: Thirty-seven samples (aqueous and vitreous) were collected from 25 eyes showing typical symptoms and clinical signs of bacterial endophthalmitis. Ocular samples were also collected from 38 eyes that underwent routine surgery and from 15 eyes with intraocular inflammation due to nonbacterial causes. Panbacterial PCR was performed with a nested pair of 16S rRNA gene primers. Subsequent bacterial identification was completed for 18 paired samples (nine eyes) using restriction fragment length polymorphism (RFLP) and DNA sequencing. RESULTS: A 100% concordance was obtained between PCR and culture-positive samples. A PCR product was amplified from all 37 intraocular samples from eyes with suspected infection, whereas only 15 of 22 vitreous samples and 5 of 15 aqueous samples were culture positive. Culture-negative PCR positive samples contained a preponderance of gram-negative bacterial sequences. Cloning and DNA analysis revealed 30 DNA sequences and included eight bacterial 16S rDNA, which currently remain unidentifiable. The presence of bacterial DNA was associated with an inflammatory response suggestive of infection and not colonization. All 15 samples from inflamed eyes with diverse uveitis diagnoses were PCR negative. The false-positive rate, due to contamination during sampling, was 5%. CONCLUSIONS: Bacterial DNA was detected in all patients with typical clinical signs of endophthalmitis. Gram-negative organisms seem to play a much more important role in the pathogenesis of this disease than previously thought. PCR-based techniques have great value in the confirmation of the diagnosis of bacterial endophthalmitis especially in culture-negative eyes. PMID- 11006242 TI - Prospective determination of T-cell responses to S-antigen in Behcet's disease patients and controls. AB - PURPOSE: To prospectively determine, using two different assays, the lymphocyte proliferative response to a retinal autoantigen (S-antigen) in patients with Behcet's disease who are under treatment for ocular inflammation. METHODS: Patients were evaluated at each visit for signs of ocular inflammation. Peripheral blood leukocytes were harvested and cultured in the presence of bovine S-antigen in a standard culture assay, as well as by limiting dilution using multiple short-term T-cell lines. RESULTS: Five patients were observed for 2 to 10 months. During follow-up, three patients had episodes of ocular inflammation. No consistent change in proliferative response was observed in standard proliferation assays. However, an increase in established T-cell lines was correlated to the presence of ocular inflammation in all three patients. Ocular activity was associated with an increase of 9- to 30-fold in the frequency of short-term T-cell lines. This increase returned to baseline within 1 to 3 months. CONCLUSIONS: An increase in S-antigen-responsive lymphocytes is found in the peripheral blood of patients with Behcet's disease during episodes of ocular inflammation. This increase cannot be measured using standard proliferation assays but requires the use of techniques exploiting the principles of limiting dilution analysis. PMID- 11006243 TI - Role and expression of CD40 on human retinal pigment epithelial cells. AB - PURPOSE: To examine the CD40 costimulatory molecule expression on normal resting or activated adult human retinal pigment epithelium (hRPE) cells and to evaluate its role as an activation molecule considering the potential antigen presentation functions of hRPE cells. METHODS: Expression of HLA-DR and costimulatory (CD40, B7.1, B7.2, CD54, and CD58) molecules on hRPE cells was analyzed by flow cytometry. CD40 triggering was performed using soluble CD40L or cocultures with CD40L transfected fibroblasts. Interleukin (IL)-6, -8, -10, and -12 secretions were measured by enzyme-linked immunosorbent assay. Antigen presentation function of hRPE cells was assessed by coculturing hRPE cells with allogeneic T cells. T cell proliferation was measured by [(3)H]-thymidine incorporation, and T-cell apoptosis by measurement of caspase-3 activity. RESULTS: Interferon (IFN)gamma activated hRPE cells expressed CD40, but not B7.1 or B7.2. Although interferongamma enhanced IL-6 and IL-8 production, CD40 triggering of IFNgamma activated hRPE cells did not induce IL-12 secretion. hRPE cells did not stimulate allogeneic resting T cells and downregulated phytohemagglutinin-activated allogeneic T cells via a cell-to-cell contact-dependent mechanism. Some induction of apoptosis was detected. CONCLUSIONS: CD40 is expressed on IFNgamma-activated hRPE cells. Its ligation leads to an increased production of IL-6 and IL-8 but fails to induce B7.1 or B7. 2 expression, or to induce IL-12 secretion. Accordingly, hRPE cells do not activate allogenic T cells but inhibit T-cell proliferation, partly through induction of apoptosis. These results suggest that hRPE cells could be implicated more in a deviant antigen presentation. If the exact molecular mechanisms are unclear, it is likely that CD40-CD40L interaction could play a role in this process. PMID- 11006245 TI - Differential expression of N- and B-cadherin during lens development. AB - PURPOSE: To analyze the dynamics of N- and B-cadherin cell adhesion molecule expression and cytoskeletal interaction during embryonic chick lens development. METHODS: Localization of N- and B-cadherin, F-actin, and connexin 56 were determined by immunohistochemistry of developing lenses or immunocytochemistry of differentiating primary lens cultures. Biochemical analysis of cytoskeletal linkage of N- or B-cadherin was assessed by differential detergent extraction, electrophoresis, and immunoblotting. RESULTS: The results indicate that although both cadherins are expressed throughout lens development, N-cadherin expression detected was similar in both lens epithelial and fiber cells, whereas B-cadherin was preferentially localized to the lens fiber cells. During differentiation, both cadherins become increasingly associated with the lens cytoskeleton, as indicated biochemically by a transition from largely Triton X-100-soluble to Triton X-100-insoluble pools and immunocytologically by cadherin localization to cell-cell borders and colocalization with the actin cytoskeleton. Although a significant fraction of N-cadherin remains Triton X-100-soluble as the lens cells differentiate, B-cadherin becomes resistant to extraction by both Triton X-100 as well as RIPA buffers. As detected immunocytochemically in lens cell cultures, the temporal localization of N-cadherin to cell-cell interfaces precedes that of B cadherin. Furthermore, temporal localization of B-cadherin, as opposed to N cadherin, to cell-cell borders more closely parallels that of connexin 56 in vitro as well as in vivo. CONCLUSIONS; These results suggest that while both N- and B-cadherin are expressed during lens cell differentiation, both their patterns of expression as well as their cytoskeletal association differ between epithelial and fiber cells. PMID- 11006244 TI - Chronic low level complement activation within the eye is controlled by intraocular complement regulatory proteins. AB - PURPOSE: To explore the role of the complement system and complement regulatory proteins in an immune-privileged organ, the eye. METHODS: Eyes of normal Lewis rats were analyzed for the expression of complement regulatory proteins, membrane cofactor protein (MCP), decay-acceleration factor (DAF), membrane inhibitor of reactive lysis (MIRL, CD59), and cell surface regulator of complement (Crry), using immunohistochemistry, Western blot analysis, and reverse transcription polymerase chain reaction (RT-PCR). Zymosan, a known activator of the alternative pathway of complement system was injected into the anterior chamber of the eye of Lewis rats. Animals were also injected intracamerally with 5 microl (25 microg) of neutralizing monoclonal antibody (mAb) against rat Crry (5I2) or CD59 (6D1) in an attempt to develop antibody induced anterior uveitis; control animals received 5 microl of sterile phosphate-buffered saline (PBS), OX-18 (25 microg), G-16 510E3 (25 microg), or MOPC-21 (25 microg). The role of complement system in antibody-induced uveitis was explored by intraperitoneal injection of 35 U cobra venom factor (CVF), 24 hours before antibody injection. Immunohistochemical staining and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Western blot analysis were used to detect the presence of membrane attack complex (MAC) and C3 activation products, respectively, in normal and antibody injected rat eyes. RESULTS: Complement activation product MAC was present in the normal rat eye, and intraocular injection of zymosan induced severe anterior uveitis. The complement regulatory proteins, MCP, DAF, CD59, and Crry, were identified in the normal rat eye. Soluble forms of Crry and CD59 were also detected in normal rat aqueous humor. Severe anterior uveitis developed in Lewis rats injected with a neutralizing mAb against Crry, with increased formation of C3 split products. Systemic complement depletion by CVF prevented the induction of anterior uveitis by anti-Crry mAb. Intracameral injection of anti-rat CD59 (6D1), anti-rat MHC class I antigen (OX-18), anti-rat Ig (G-16-510E3), or MOPC-21 caused no inflammatory reaction. CONCLUSIONS: The results suggest that the complement system is continuously active at a low level in the normal eye and is tightly regulated by intraocular complement regulatory proteins. PMID- 11006246 TI - A nonsense mutation (W9X) in CRYAA causes autosomal recessive cataract in an inbred Jewish Persian family. AB - PURPOSE: To identify the genetic defect causing autosomal recessive cataract in two inbred families. METHODS: Linkage analysis was performed with polymorphic markers close to 14 loci previously shown to be involved in autosomal dominant congenital cataract. In one of the families a gene segregating with the disease was analyzed by single-strand conformation polymorphism (SSCP) and eventually sequenced. RESULTS: Three polymorphic markers close to the CRYAA gene located on chromosome 21q segregated with the disease phenotype in one of the families, but not in the other. Sequencing of the CRYAA in this Jewish Persian family revealed a G-to-A substitution, resulting in the formation of a premature stop codon (W9X). CONCLUSIONS: A nonsense mutation in the CRYAA gene causes autosomal recessive cataract in one family. This constitutes the first description of the molecular defect underlying nonsyndromic autosomal recessive congenital cataract. That there was no linkage to this locus in another family provides evidence for genetic heterogeneity. PMID- 11006247 TI - Effects of experimentally induced ametropia on the morphology and optical quality of the avian crystalline lens. AB - PURPOSE: To examine the effects of refractive error on avian lens morphology and optical quality. METHODS: Hatchling white leghorn chicks were unilaterally goggled for 7 days with either a form-deprivation goggle (n = 12), a -10 D defocus goggle (n = 12), or a +10 D defocus goggle (n = 12) to induce myopia and hyperopia. Optical quality of lenses (focal length and focal length variability) from treated and contralateral control eyes was assessed using a scanning laser apparatus. Lens morphology was examined by light and electron microscopy. RESULTS: Although the induction of refractive errors did not significantly alter lens size, shape, paraxial focal length, or average focal length, average focal length variability increased. Lenses from eyes goggled with form-deprivation and +10 D defocus goggles demonstrated a twofold increase in average focal length variability, when compared with their contralateral controls. The morphology of the lens is not altered by these experimental manipulations. CONCLUSIONS: This study provides evidence that the refractive development of the chick lens is not independent of the refractive development of the ocular globe and that chick lenticular development is influenced by both genetics and visual experience. PMID- 11006249 TI - Influence of exposure time for UV radiation-induced cataract. AB - PURPOSE: It is believed that for a certain ultraviolet radiation (UVR) exposure, the biologic effect depends on the product of irradiance and exposure time (the reciprocity Bunsen-Roscoe law). The purpose of this study was to investigate the validity of the reciprocity law for UVR-induced cataract. METHODS: Two experiments were conducted. In the first one, 100 Sprague-Dawley rats were exposed to UVR divided into five groups according to exposure time: 7.5, 15, 30, 60, and 120 minutes. In the second experiment, 80 Sprague-Dawley rats were exposed to UVR divided into four groups according to exposure time: 5, 7.5, 11, and 15 minutes. All the animals were unilaterally exposed to the same dose of UVR (8 kJ/m(2)) in the 300-nm wavelength region. One week after exposure both lenses were removed to measure the intensity of forward light scattering and for microphotography. Groups were compared by evaluating the difference between exposed and nonexposed eyes. RESULTS: The group exposed to UVR for 5 minutes had the lowest intensity of forward light scattering. The highest intensity of forward light scattering was found in the group that was exposed for 15 minutes. With longer exposure intervals, the intensity of forward light scattering decreased as the exposure time increased. No difference in intensity of forward light scattering was found between the groups exposed for 60 and 120 minutes. CONCLUSIONS; Exposure time strongly influenced cataract formation after low-dose UVR. In this model of UVR-induced cataract, the photochemical reciprocity law was modulated by a biologic response. PMID- 11006248 TI - DeltaFosB-induced cataract. AB - PURPOSE: The objective of this study was to investigate a possible relationship between posterior subcapsular cataract (PSC) formation and expression of the transcription factor DeltaFosB. METHODS: Western blot analysis was performed on bitransgenic NSE-tTA, TetOp-DeltaFosB, and single-transgenic NSE-tTA control mice to determine the pattern of DeltaFosB expression within the eye. Light and scanning electron microscopy and biochemical analyses were also performed. RESULTS: In mice expressing DeltaFosB, cataract developed that initially appeared to be posterior subcapsular and gradually matured to involve the entire lens. The enlarged posterior ends of developing secondary fibers curved away from the visual axis to form an elevated opaque posterior plaque. As a result, posterior suture formation did not occur. At a later time, the attenuated posterior capsule overlying the plaque ruptured and the lens nucleus subluxated into the vitreous. Retinal damage was also observed but only from postnatal day 65, a time when extensive lens degeneration had already occurred. DeltaFosB expression was observed well before the detection of morphologic change in both the lens and the retina. Within the lens, DeltaFosB expression was found in both the epithelium and fibers. The development of cataracts was a direct consequence of DeltaFosB expression and was not due to the disruption of an endogenous gene by transgene integration since cataracts could be prevented by silencing expression of DeltaFosB by feeding bitransgenic animals doxycycline (Dox). Moreover, cataracts were observed in bitransgenic mice derived from two independent TetOp-DeltaFosB founder lines but not in single NSE-tTA transgenic controls. Cataractogenesis was not a consequence of abnormal development, because mice conceived and raised on Dox to prevent expression of DeltaFosB also were subject to formation of PSC when expression of DeltaFosB was turned on in adult animals by removing Dox. Examination of biochemical parameters indicated that the earliest change observed was the disruption of calcium homeostasis with a significant increase in Ca(2+) influx, followed by a gradual but marked decrease in protein content. Significant changes in certain metabolic parameters and protein composition were also observed. CONCLUSIONS: The DeltaFosB-induced cataract in which the major morphologic early event was the disruption of normal posterior fiber formation, may be a good model for PSC. By identifying DeltaFosB-regulated target genes, it should be possible to achieve a better understanding of the molecular mechanisms through which PSC is formed. PMID- 11006250 TI - Susceptibility to TGFbeta2-induced cataract increases with aging in the rat. AB - PURPOSE: Cataract is the most common cause of blindness in the world today, and yet there is no generally accepted treatment other than surgical intervention. Studies in rodent models designed to increase understanding of the molecular basis of cataract have shown that transforming growth factor (TGF)-beta induces morphologic and molecular changes similar to those associated with some forms of human cataract. Because aging is the most widely recognized risk factor for cataract, it is important that any animal model be examined in this context. This was a study of the effects of aging on susceptibility to TGFbeta-induced cataract. METHODS: Lenses from weanling, adult, and senile rats were cultured in defined serum-free medium with a range of concentrations of TGFbeta2. The lenses were cultured for up to 7 days, photographed daily, fixed, and prepared for histology and immunolocalization. Opacification was quantified by image analysis. RESULTS: Lenses from weanling, adult, and senile rats all underwent similar cataractous changes when exposed to TGFbeta. This included opacification, the formation of anterior subcapsular plaques, and accumulation of type I collagen and alpha-smooth muscle actin. Lenses from adult and senile animals, however, were generally more adversely affected by TGFbeta than lenses from weanlings. This study also showed that a low dose of TGFbeta administered over a prolonged period had an effect similar to that of a higher dose administered over a shorter period. CONCLUSIONS: An elevation of TGFbeta activity, either acute or chronic, and/or an age-related increase in lens cell susceptibility to TGFbeta may be triggering factors in the etiology of certain forms of cataract. PMID- 11006251 TI - Methazolamide 1% in cyclodextrin solution lowers IOP in human ocular hypertension. AB - PURPOSE: To formulate aqueous eye drops containing methazolamide 1% in cyclodextrin solution and to evaluate their effect on intraocular pressure (IOP) in a double-blind randomized trial in humans. Methazolamide, a carbonic anhydrase inhibitor (CAI), has been used in oral doses in the treatment of glaucoma but hitherto has not been successfully formulated in eye drops. In this study the effects of methazolamide are compared with those of dorzolamide (Trusopt). METHODS: Methazolamide 1% was formulated in a 2-hydroxypropyl-beta-cyclodextrin with hydroxypropyl methylcellulose in aqueous solution. Eight persons with ocular hypertension were treated with the methazolamide-cyclodextrin eye drops and eight persons with dorzolamide (Trusopt), both groups at dosages of three times a day for 1 week. IOP was measured before treatment was begun and on days 1, 3, and 8 at 9 AM (peak) and 3 PM (trough). RESULTS: After 1 week of treatment, the peak IOP in the methazolamide group had decreased from 24.4 +/- 2.1 mm Hg (mean +/- SD) to 21.0 +/- 2.0 mm Hg, which is a 14% pressure decrease (P: = 0.006). In the dorzolamide group, the peak IOP decreased from 23.3 +/- 2.1 mm Hg to 17.2 +/- 3.1 mm Hg, which is a 26% pressure decrease (P: < 0.001). On average, the IOP declined 3.4 +/- 1.8 mm Hg after methazolamide administration and 6.1 +/- 3.6 mm Hg after dorzolamide. CONCLUSIONS: Through cyclodextrin complexation, it is possible to produce topically active methazolamide eye drops that lower IOP. This is the first double-blind clinical trial that demonstrates the efficacy of the classic CAIs in eye drop formulation. PMID- 11006252 TI - A retinal-derived relaxing factor mediates the hypoxic vasodilation of retinal arteries. AB - PURPOSE: To investigate the mechanisms involved in hypoxic vasodilation using an in vitro setup. METHODS: Retinal arteries with and without retinal tissue were mounted on a wire myograph. The segments were contracted with prostaglandin (PG)F(2alpha) (30 microM) or 120 mM K(+). Hypoxia was induced by replacement of O(2) by N(2) in the gas used to bubble the Krebs-Ringer bicarbonate organ bath solution. RESULTS: Hypoxia induced complete relaxation of preparations with adherent retinal tissue contracted with PGF(2alpha). Preparations without retinal tissue were not affected by the change in oxygenation. When the retinal arteries were contracted with 120 mM K(+), hypoxia no longer induced relaxation of the preparation with adherent retinal tissue. The presence of an NO-synthase inhibitor (L-NA, 0.1 mM), a cyclooxygenase inhibitor (indomethacin, 50 microM), or an adenosine receptor antagonist (8-sulfophenyltheophylline, 1 mM) did not affect hypoxic vasodilation. Excitatory amino acids and lactate had no or only a limited effect on the PGF(2alpha)-induced contraction and are therefore unlikely mediators of hypoxic vasodilation. HCl (10 mM) reduced the pH to 6.1 +/- 0.08 (n = 4) and induced a pronounced but transient relaxation of the retinal artery contracted with PGF(2alpha) or 120 mM K(+), whereas hypoxia induced relaxation of the retinal artery contracted with PGF(2alpha) only in the presence of adherent retinal tissue. CONCLUSIONS: Adherent retinal tissue mediates the hypoxic vasodilatation of bovine retinal arteries in vitro. Neither NO, prostanoids, adenosine, excitatory amino acids lactate or changes in pH seem to be involved in this hypoxic response. PMID- 11006253 TI - Altered expression of retinal occludin and glial fibrillary acidic protein in experimental diabetes. The Penn State Retina Research Group. AB - PURPOSE: To investigate how diabetes alters vascular endothelial cell tight junction protein and glial cell morphology at the blood-retinal barrier (BRB). METHODS: The distribution of the glial marker, glial fibrillary acidic protein (GFAP), and the endothelial cell tight junction protein occludin were explored by immunofluorescence histochemistry in flatmounted retinas of streptozotocin (STZ) diabetic and age-matched control rats, and in BB/Wor diabetes-prone and age matched diabetes-resistant rats. RESULTS: GFAP immunoreactivity was limited to astrocytes in control retinas. Two months of STZ-diabetes reduced GFAP immunoreactivity in astrocytes and increased GFAP immunoreactivity in small groups of Muller cells. After 4 months of STZ-induced diabetes, all Muller cells had intense GFAP immunoreactivity, whereas there was virtually none in the astrocytes. BB/Wor diabetic rats had similar changes in GFAP immunoreactivity. Occludin immunoreactivity in normal rats was greatest in the capillary bed of the outer plexiform layer and arterioles of the inner retina but much less intense in the postcapillary venules. Diabetes reduced occludin immunoreactivity in the capillaries and induced redistribution from continuous cell border to interrupted, punctate immunoreactivity in the arterioles. Forty-eight hours of insulin treatment reversed the pattern of GFAP and occludin immunoreactivity in the STZ-diabetic rats. CONCLUSIONS: Diabetes alters GFAP expression in retinal glial cells, accompanied by reduction and redistribution of occludin in endothelial cells. These changes are consistent with the concept that altered glial-endothelial cell interactions at the BRB contribute to diabetic retinopathy. PMID- 11006254 TI - Safety and pharmacokinetics of an intraocular fluocinolone acetonide sustained delivery device. AB - PURPOSE: To determine the safety and pharmacokinetics of an intraocular fluocinolone acetonide sustained drug delivery device. METHODS: Nonbiodegradable drug delivery devices containing 2 or 15 mg of a synthetic corticosteroid, fluocinolone acetonide, were constructed. The long-term in vitro release rates of these devices were determined in protein-free buffer or buffer containing 50% plasma protein. Fifteen-milligram devices were also implanted into the vitreous cavities of rabbit eyes. Intravitreal drug levels, the amount of drug remaining in explanted devices, and the release rate of explanted devices were determined over a 1-year time period. Drug toxicity was assessed over this same time period by slit lamp examination, indirect ophthalmoscopy, electroretinography, and histologic examination. RESULTS: The drug release rates for the 2-mg device, 1.9 +/- 0.25 microg/d, and for the 15-mg device, 2.2 +/- 0.6 microg/d, remained linear over the 6-month and 45-day testing period, respectively. The release rate increased by approximately 20% when devices were transferred from protein-free buffer to buffer that contained protein (P: < 0.0001). Vitreous levels remained fairly constant (0.10-0.21 microg/ml) over a 1-year period. No drug was present in the aqueous humor during this time period. Based on the device release rates, the predicted life span of the 2- and 15-mg devices are 2.7 and 18.6 years, respectively. There was no evidence of drug toxicity by clinical examination, electroretinography, or histologic examination. CONCLUSIONS: It is feasible to construct a nontoxic fluocinolone acetonide drug delivery device that reproducibly releases fluocinolone acetonide in a linear manner over an extended period. These devices show great promise in the treatment of ocular diseases such as uveitis, which are often managed with chronic corticosteroid therapy. PMID- 11006255 TI - Elevation of vitreous leptin in diabetic retinopathy and retinal detachment. AB - PURPOSE: Leptin is a cytokine that regulates energy metabolism and is linked to diabetes mellitus through its metabolic actions. Leptin is angiogenic and promotes wound healing, and therefore this investigation was conducted to determine whether leptin is associated with neovascular and fibrotic complications of diabetes and other retinopathies. METHODS: Serum and vitreous samples were collected from patients classified by the presence and type of diabetic retinopathy or other ocular diseases. Leptin was measured in serum and vitreous by radioimmunoassay, and leptin and leptin receptor were localized in epiretinal membranes immunohistochemically. RESULTS: Leptin levels in serum and vitreous were higher in patients with diabetes than in those without, and vitreous leptin concentrations were especially elevated in patients with proliferative diabetic retinopathy or retinal detachment. Leptin and leptin receptor were detected in fibrovascular epiretinal membrane of patients with diabetes. CONCLUSIONS: Leptin in human vitreous is elevated in proliferative diabetic retinopathy, and retinal detachment and is present in fibrovascular epiretinal tissue. These data suggest an involvement of leptin in retinal disease. PMID- 11006256 TI - Choroidal neovascularization in the rat induced by adenovirus mediated expression of vascular endothelial growth factor. AB - PURPOSE: To determine the effects of an adenovirus vector encoding vascular endothelial growth factor(165) (Ad.VEGF) delivered to the subretinal space in the rat. METHODS: An E1-deleted adenoviral vector encoding VEGF was injected into the subretinal space of Long-Evans rats. Immunohistochemistry identified VEGF expression. Histopathologic changes in the retina were determined by light and electron microscopy, immunohistochemistry, fluorescein angiography, and examination of wholemounts of choroid and retina. RESULTS: Increased expression of VEGF only in the retinal pigment epithelium (RPE) was detected after Ad.VEGF injection. Histopathology of these eyes revealed minimal subretinal exudation at 1 week followed by the appearance of vascular structures in the subretinal space by week 2, which persisted up to 4 weeks. Shortening of photoreceptor outer segments and reduction of the outer nuclear layer were present overlying areas of neovascularization. Fluorescein angiography of animals injected with fluorescein dextran revealed a deep complex of new vessels. Choroidal flatmounts showed new vessel formation, verified by detection of endothelial cells via immunohistochemistry, arising from the choroid with absence of change in the overlying retinal vasculature. Electron microscopy confirmed the presence of sub RPE endothelial cells and pericytes and the loss of integrity of Bruch's membrane, and serial sectioning demonstrated choroidal vascular growth through Bruch's membrane. CONCLUSIONS: These results support the hypothesis that overexpression of VEGF from RPE cells is capable of inducing choroidal neovascularization in the rat and provide a framework for further examining angiogenic processes in the RPE-choroid complex. PMID- 11006257 TI - Coordination between production and turnover of interphotoreceptor retinoid binding protein in zebrafish. AB - PURPOSE: Interphotoreceptor retinoid-binding protein (IRBP), which is secreted by the photoreceptors of most vertebrates, is the major soluble protein component of the interphotoreceptor matrix (IPM). Recent studies suggest that IRBP is short lived in the IPM (half-life, approximately 11 hours). The mechanisms coordinating the production and removal of IRBP are not known. Zebrafish provide a useful system to study the regulation of these two processes, because its IRBP mRNA levels are under circadian regulation. In the present study, the relationship between the quantity of IRBP, the rate of its turnover, and the expression of its mRNA in the zebrafish retina were examined. METHODS: Full-length zebrafish IRBP was expressed in Escherichia coli and an antiserum generated against purified recombinant IRBP. Western and protein dot blot analyses and indirect immunofluorescence were used to define the temporal and spatial patterns of IRBP expression in the adult zebrafish. In vivo and in vitro metabolic labeling experiments were used to examine the regulation of IRBP turnover by both environmental light and the light-dark cycle. RESULTS: Despite the known rhythmicity in IRBP mRNA expression, neither the amount of IRBP nor its localization changes significantly during the light-dark cycle. IRBP is rapidly removed from the zebrafish eye (half life, approximately 7 hours). This rapid turnover is independent of environmental lighting conditions during subjective day and is more rapid during the day than at night. CONCLUSIONS: Because the amount of IRBP remains constant throughout the day, the enhanced daytime IRBP mRNA expression may function to compensate for an increased turnover of the protein during the day. These findings suggest that the processes of IRBP production and removal are coordinately regulated. PMID- 11006258 TI - Distribution and developmental regulation of AMPA receptor subunit proteins in rat retina. AB - PURPOSE: To learn more about a possible functional role of alpha-amino-3-hydroxy 5-methyl-4-isoxasole-propionate (AMPA) receptors in retinal development, the spatial distribution and temporal regulation of all AMPA receptor subunit proteins was studied in rats. METHODS: Immunohistochemistry was performed on retinal sections between embryonic days (E)20 and E21 and the adult stage by using specific antibodies against AMPA subunits GluR1 to 4. RESULTS: All AMPA subunits were expressed in the ganglion cell layer from E21 on. In the inner plexiform layer (IPL), discernible bands of labeling appeared at distinct retinal ages for the different subunits. GluR1 immunoreactivity (IR) was concentrated in two broad bands by postnatal day (P)3, whereas three bands were visible beginning on P9. Two bands were located in a region of the IPL where off-cells terminate, and one band was found in the innermost part of the IPL where on-cells terminate. In contrast, two bands of GluR2/3- and GluR4-IR in the IPL were only discernible beginning on P14 and seemed to be located between the bands of GluR1-IR. GluR2/3 and GluR4 were observed both in horizontal cells and in the outer plexiform layer from early developmental stages on. GluR1 was not found in the outer retina, indicating that horizontal and bipolar cell processes in the rat express AMPA receptors composed of subunits GluR2 to 4. Double-labeling experiments with cell specific markers revealed the expression of subunits GluR1 to 4 in cholinergic and AII amacrine cells. CONCLUSIONS: AMPA receptors are expressed before synapse formation, indicating a role not only in fast signal transmission but also in the establishment of inner retinal circuits. The differences in spatial and temporal subunit expression suggest that different retinal cell types selectively express distinct types of AMPA receptors during development of the rat retina. PMID- 11006259 TI - Lipid peroxidation and peroxynitrite in retinal ischemia-reperfusion injury. AB - PURPOSE: To investigate whether lipid peroxides play a role in retinal cell death due to ischemia-reperfusion injury, whether recombinant human thioredoxin (rhTRX) treatment reduces production of lipid peroxides of the retina, and whether such treatment reduces the number of cells expressing c-Jun and cyclin D1. METHODS: Retinal ischemia was induced in rats by increasing the intraocular pressure to 110 mm Hg for 60 minutes. After reperfusion, immunohistochemical staining for lipid peroxide, peroxynitrite, c-Jun, and cyclin D1 and propidium iodide (PI) staining were performed on retinal sections from animals treated intravenously with and without rhTRX, a free radical scavenger. Quantitative analyses of PI-, c Jun-, and cyclin D1-positive cells were performed after the ischemic insult. Concentration of lipid peroxides in the retina was determined by the thiobarbituric acid assay. RESULTS: Specific immunostaining for lipid peroxides was seen in the ganglion cell layer at 6 hours after reperfusion, in the inner nuclear layer at 12 hours, and in the outer nuclear layer at 48 hours. Time course studies for PI-positive cells in the three nuclear layers coincided with those of specific immunostaining for lipid peroxides. The specific immunostaining was weakened by pre- and posttreatment with 0.5 mg of rhTRX. The number of PI-, c Jun-, and cyclin D1-positive cells and the concentration of lipid peroxides were significantly decreased by treatment with rhTRX compared with those of vehicle treated control rats (P: < 0. 01). CONCLUSIONS: Lipid peroxides formed by free radicals may play a role in neuronal cell death in retinal ischemia-reperfusion injury. PMID- 11006261 TI - Retinal degeneration is slowed in transgenic rats by AAV-mediated delivery of FGF 2. AB - PURPOSE: We evaluated adeno-associated virus (AAV)-mediated gene transfer of basic fibroblast growth factor (FGF-2) as a therapy for photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa. METHODS: Recombinant adeno-associated virus vector (rAAV) incorporating a constitutive cytomegalovirus (CMV) promoter was used to transfer the bovine FGF-2 gene to photoreceptors. AAV was administered by subretinal injection to transgenic rats (TgN S334ter-4) at postnatal day 15 (P15). Control eyes were uninjected, injected with PBS, or AAV-LacZ. Eyes were examined by histopathology, morphometric analysis, and electroretinography at P60. RESULTS: Expression of recombinant FGF 2 slowed the rate of photoreceptor degeneration. Morphologic studies demonstrated significantly more photoreceptors surviving in eyes injected with AAV-FGF-2 than in controls. Insignificant rescue effects were seen in retinas injected with buffer only. No significant inflammatory response or neovascularization was detected. Electroretinographic (ERG) responses of eyes injected with AAV-FGF-2 were increased compared with uninjected eyes; however, these amplitudes were not significantly larger than eyes receiving an AAV-LacZ control vector. CONCLUSIONS: Transduction of retinal cells with AAV-FGF-2 reduces the rate of photoreceptor degeneration in an S334ter-4 animal model. Despite the lack of significantly increased ERG amplitudes from eyes expressing FGF-2, a greater number of surviving photoreceptors was demonstrated. Delivery of FGF-2 using recombinant AAV has potential as a therapy for retinal degeneration. PMID- 11006260 TI - Depression of retinal glutamate transporter function leads to elevated intravitreal glutamate levels and ganglion cell death. AB - PURPOSE: Elevated levels of extracellular glutamate have been implicated in the pathophysiology of neuronal loss in both central nervous system and ophthalmic disorders, including glaucoma. This increase in glutamate may result from a failure of glutamate transporters (molecules that ordinarily regulate extracellular glutamate; E:xcitatory A:mino A:cid T:ransporter; EAAT). Elevated glutamate levels can also lead to alterations in glutamate receptor expression. It was hypothesized that selective blockade of glutamate transporters would be toxic to retinal ganglion cells. METHODS: Glutamate transporters were blocked either pharmacologically or with subtype-specific antisense oligonucleotides against EAAT1. Glutamate levels, transporter levels and ganglion cell survival were assayed. RESULTS: Pharmacological inhibition of glutamate transporters with either an EAAT2 specific inhibitor or a nonspecific inhibitor of all the subtypes of transporters was toxic to ganglion cells. Treatment with oligonucleotides against the glutamate transporter EAAT1 decreased the levels of expression of the transporter, increased vitreal glutamate, and was toxic to ganglion cells. CONCLUSIONS: These results demonstrate that normal function of EAAT1 and EAAT2 is necessary for retinal ganglion cell survival and plays an important role in retinal excitotoxicity. Manipulation of retinal glutamate transporter expression may become a useful tool in understanding retinal neuronal loss. PMID- 11006262 TI - Effects of hypoxemia on the a- and b-waves of the electroretinogram in the cat retina. AB - PURPOSE: Slow components of the electroretinogram (ERG) are sensitive to even mild hypoxemia (60 < P(a)O(2) < 100 mm Hg) in the cat eye. However, the electrical responses of the inner retina remain unchanged until P(a)O(2) is below 40 mm Hg. In this study, the effects of hypoxemia on photoreceptors, on which both slow ERG components and inner retinal activity depend, were examined by recording the a-wave of the ERG. METHODS: The ERG of dark-adapted, anesthetized cats was recorded between an Ag-AgCl electrode in the vitreous humor and a reference electrode near the eye. Responses to bright flashes of diffuse white light were recorded at 3-minute intervals during hypoxemic episodes lasting 15 minutes to 2 hours. RESULTS: The cat a-wave was well described by the Lamb and Pugh a-wave model during normoxia and hypoxemia. During mild hypoxemia (P(a)O(2) of 50-60 mm Hg), small changes in a-wave amplitude were detected but did not become greater during severe hypoxemia. The mean decrease in the a-wave amplitude during severe hypoxemia (P(a)O(2) of 20-30 mm Hg) was 8.9% from the mean amplitude during air breathing. The effects of hypoxemia were more severe on the b-wave amplitude. The mean decrease in the b-wave was 35% at P(a)O(2) of 20-30 mm Hg. CONCLUSIONS: The a-wave is more resistant to severe hypoxemia than the b wave. This implies that photoreceptor transduction works almost normally during hypoxemia and that failure of inner retinal PO(2) regulation causes the decrease in the b-wave. Previously observed changes in the amplitudes of slow ERG components during hypoxemia may result from changes in the ionic environment, rather than a failure of photoreceptor energy metabolism. PMID- 11006263 TI - The nature and extent of retinal dysfunction associated with diabetic macular edema. AB - PURPOSE: To evaluate the nature and extent of retinal dysfunction in the macular and surrounding areas that occurs in patients with diabetes with clinically significant macular edema (CSME). METHODS: Eleven patients were evaluated before focal laser treatment. Multifocal electroretinogram (ERG) and full-field ERG techniques were used to assess the effects of diabetic retinopathy and CSME on macular, paramacular, and peripheral retinal function. A modified visual field technique was used to obtain local threshold fields. The relationship between local sensitivity changes and local ERG changes was determined. RESULTS: Local ERG responses were significantly delayed and decreased in amplitude, and timing changes were observed in a larger area of the retina than amplitude changes. Visual field deficits were similarly widespread with marked sensitivity losses occurring in retinal areas with normal ERG amplitudes and in areas that appeared to be free of fundus abnormalities. Despite this similarity and the finding that retinal areas with elevated thresholds have timing delays, timing delays were not good predictors of the degree of threshold elevation. CONCLUSIONS: The results demonstrate the widespread nature of timing deficits and visual field deficits that are associated with CSME. PMID- 11006265 TI - Defining ocular diseases PMID- 11006264 TI - Retinal function in diabetic macular edema after focal laser photocoagulation. AB - PURPOSE: To assess the effects of focal photocoagulation on retinal function in the macular and perimacular areas in patients with diabetes who have clinically significant macular edema. METHODS: Eleven patients were assessed after focal laser treatment. Multifocal electroretinogram (ERG) and full-field ERG techniques were used to evaluate the effects of treatment on macular, paramacular, and peripheral retinal function. A modified visual field technique was used to obtain local threshold fields. The posttreatment results were compared with pretreatment results. Changes in local ERG response amplitudes and implicit times were calculated for each patient and presented as difference fields. The changes in local ERG responses were compared with the changes in local field sensitivity. RESULTS: After treatment, the results of the psychophysical tests suggested little or no change in visual function, but changes in retinal function were observed with the multifocal ERG technique. Local ERG responses showed increases in implicit time and decreases in amplitude, compared with pretreatment values. Timing was affected more than amplitude. CONCLUSIONS: The results suggest that focal treatment produces changes in retinal function, and these changes are not restricted to the treated macular area. PMID- 11006266 TI - XIST RNA associates with specific regions of the inactive X chromatin. AB - Microscopy studies have shown that XIST RNA colocalizes with the inactive X chromosome (Xi). However, the molecular basis for this colocalization is unknown. Here we provide two lines of evidence from chromatin immunoprecipitation experiments that XIST RNA physically associates with the Xi chromatin. First, XIST RNA can be co-precipitated by antiserum against macroH2A, a histone H2A variant enriched in the Xi. Second, XIST RNA can be co-precipitated by antisera that recognize unacetylated, but not acetylated, isoforms of histones H3 and H4. The specificity of XIST RNA association with hypoacetylated chromatin, together with the previous finding that hypoacetylated histone H4 is enriched at promoters of X-inactivated genes, raises the possibility that XIST RNA may contribute to the hypoacetylation of specific regions of the Xi so as to alter the expression of X-linked genes. PMID- 11006267 TI - Chemical rescue of a mutant protein-tyrosine kinase. AB - Protein-tyrosine kinases contain a catalytic loop Arg residue located either two or four positions downstream of a highly conserved Asp residue. In this study, the role of this Arg (Arg-318) in the protein-tyrosine kinase C-terminal Src kinase (Csk) was investigated. The observed k(cat) for phosphorylation of the random copolymer poly(Glu,Tyr) substrate by Csk R318A is approximately 3000-fold smaller compared with that of wild type Csk, whereas the K(m) values for ATP and poly(Glu,Tyr) are only mildly affected. The k(cat) value for poly(Glu,Tyr) phosphorylation by the Csk double mutant A316R,R318A is 100-fold greater than the k(cat) value for the single R318A mutant, suggesting that an Arg positioned at the alternative location fulfills a similar function as in wild type. Csk R318A kinase activity can also be partially recovered by several exogenous small molecules including guanidinium and imidazole. These molecules contain key features whose roles in catalysis can be rationalized from a known x-ray structure of the insulin receptor tyrosine kinase. Imidazole is the best of these activators, enhancing phosphorylation rates by Csk R318A up to 100-fold for poly(Glu,Tyr) and significantly stimulating Csk R318A phosphorylation of the physiologic substrate Src. This chemical rescue of mutant protein kinase activity might find applications in cell signal transduction experiments. PMID- 11006268 TI - Thyroid-stimulating hormone and cyclic AMP activate p38 mitogen-activated protein kinase cascade. Involvement of protein kinase A, rac1, and reactive oxygen species. AB - p38 mitogen-activated protein kinases (p38-MAPKs) are activated by cytokines, cellular stresses, growth factors, and hormones. We show here that p38-MAPKs are activated upon stimulation by thyroid-stimulating hormone (TSH) or cAMP. TSH caused the phosphorylation of p38-MAPK in Chinese hamster ovary cells stably transfected with the human TSH receptor but not in wild-type Chinese hamster ovary cells. The effect of TSH was fully mimicked by the adenylyl cyclase activator, forskolin, and by a permeant analog of cAMP. The effect of forskolin was reproduced in FRTL5 rat thyroid cells. TSH also stimulated the phosphorylation of MAPK kinase 3 or 6, over the same time scale as that of p38 MAPKs. TSH and forskolin stimulated the activity of the alpha-isoform of p38-MAPK assayed by phosphorylation of the transcription factor ATF2. The activity of MAPK activated protein kinase-2 was stimulated by TSH and forskolin. This stimulation was abolished by SB203580, a specific inhibitor of p38-MAPKs. The protein kinase A inhibitor H89 inhibited the stimulation of phosphorylation of p38-MAPKs by forskolin, whereas inhibitors of protein kinase C, p70(S6k), and phosphatidylinositol 3-kinase were ineffective. Expression of the dominant negative form of Rac1, but not that of Ras, blocked forskolin-induced p38-MAPK activation. Diphenylene iodonium, a potent inhibitor of NADPH oxidase(s), and ascorbic acid, an effective free radical scavenger, suppressed TSH- or forskolin stimulated p38-MAPK phosphorylation, indicating that the generation of reactive oxygen species plays a key role in signaling from cAMP to p38-MAPKs. Inhibition of the p38-MAPK pathway with SB203580 partially but significantly, attenuates cAMP- and TSH-induced expression of the sodium iodide symporter in FRTL-5 cells. These results point to a new signaling pathway for the G(s)-coupled TSH receptor, involving cAMP, protein kinase A, Rac1, and reactive oxygen species and resulting in the activation of a signaling kinase cascade that includes MAPK kinase 3 or 6, p38-MAPK, and MAPK-activated protein kinase-2. PMID- 11006269 TI - Regions of prostate-specific antigen (PSA) promoter confer androgen-independent expression of PSA in prostate cancer cells. AB - Prostate-specific antigen (PSA) is expressed primarily by both normal prostate epithelium and the vast majority of prostate cancers. Increases in serum PSA during endocrine therapy are generally considered as evidence for prostate cancer recurrence or progression to androgen independence. The mechanisms by which PSA up-regulation occurs in androgen-refractory prostate cancer cells are unknown. In this study, by using LNCaP and its lineage-derived androgen-independent PSA producing subline, C4-2, we identified two cis-elements within the 5.8-kilobase pair PSA promoter that are essential for the androgen-independent activity of PSA promoter in prostate cancer cells. First, a previously reported 440-bp androgen responsive element enhancer core (AREc) was found to be important for the high basal PSA promoter activity in C4-2 cells. Both mutation analysis and supershift experiments demonstrated that androgen receptor (AR) binds to the AREs within the AREc and activate the basal PSA promoter activity in C4-2 cells under androgen deprived conditions. Second, a 150-bp pN/H region was demonstrated to be a strong AR-independent positive-regulatory element of the PSA promoter in both LNCaP and C4-2 cells. Through DNase I footprinting and linker scan mutagenesis, a 17-bp RI site was identified as the key cis-element within the pN/H region. Data from electrophoretic mobility shift analysis and UV cross-linking experiments further indicated that a 45-kDa (p45) cell-specific transcription factor associates with RI in prostate cancer cells and may be responsible for driving the PSA promoter activity independent of androgen and AR. Furthermore, by juxtaposing AREc and pN/H, we produced a chimeric PSA promoter (supra-PSA) that exhibits 2-3-fold higher activity than the wild type PSA promoter in both LNCaP and C4-2 cells. PMID- 11006270 TI - A mutant Escherichia coli tyrosyl-tRNA synthetase utilizes the unnatural amino acid azatyrosine more efficiently than tyrosine. AB - Alloproteins, proteins that contain unnatural amino acids, have immense potential in biotechnology and medicine. Although various approaches for alloprotein production exist, there is no satisfactory method to produce large quantities of alloproteins containing unnatural amino acids in specific positions. The tyrosine analogue azatyrosine, l-beta-(5-hydroxy-2-pyridyl)-alanine, can convert the ras transformed phenotype to normal phenotype, presumably by its incorporation into cellular proteins. This provided the stimulus for isolation of a mutant tyrosyl tRNA synthetase (TyrRS) capable of charging azatyrosine to tRNA. A plasmid library of randomly mutated Escherichia coli tyrS (encoding TyrRS) was made by polymerase chain reaction techniques. The desired TyrRS mutants were selected by screening for in vivo azatyrosine incorporation of E. coli cells transformed with the mutant tyrS plasmids. One of the clones thus isolated, R-6-A-7, showed a 17 fold higher in vivo activity for azatyrosine incorporation than wild-type TyrRS. The mutant tyrS gene contained a single point mutation resulting in replacement of phenylalanine by serine at position 130 in the protein. Structural modeling revealed that position 130 is located close to Asp(182), which directly interacts with tyrosyladenylate. Kinetic analysis of aminoacyl-tRNA formation by the wild type and mutated F130S TyrRS enzymes showed that the specificity for azatyrosine, measured by the ratios of k(cat)/K(m) for tyrosine and the analogue, increased from 17 to 36 as a result of the F130S mutation. Thus, the high discrimination against azatyrosine is significantly reduced in the mutant enzyme. These results suggest that utilization of F130S TyrRS for in vivo protein biosynthesis may lead to efficient production of azatyrosine-containing alloproteins. PMID- 11006272 TI - A novel hepatointestinal leukotriene B4 receptor. Cloning and functional characterization. AB - Leukotriene B(4) (LTB(4)) is a product of eicosanoid metabolism and acts as an extremely potent chemotactic mediator for inflammation. LTB(4) exerts positive effects on the immigration and activation of leukocytes. These effects suggest an involvement of LTB(4) in several diseases: inflammatory bowel disease, psoriasis, arthritis, and asthma. LTB(4) elicits actions through interaction with one or more cell surface receptors that lead to chemotaxis and inflammation. One leukotriene B(4) receptor has been recently identified (LTB(4)-R1). In this report we describe cloning of a cDNA encoding a novel 358-amino acid receptor (LTB(4)-R2) that possesses seven membrane-spanning domains and is homologous (42%) and genetically linked to LTB(4)-R1. Expression of LTB(4)-R2 is broad but highest in liver, intestine, spleen, and kidney. In radioligand binding assays, membranes prepared from COS-7 cells transfected with LTB(4)-R2 cDNA displayed high affinity (K(d) = 0.17 nm) for [(3)H]LTB(4). Radioligand competition assays revealed high affinities of the receptor for LTB(4) and LTB(5), and 20-hydroxy LTB(4), and intermediate affinities for 15(S)-HETE and 12-oxo-ETE. Three LTB(4) receptor antagonists, 14,15-dehydro-LTB(4), LTB(4)-3-aminopropylamide, and U 75302, had high affinity for LTB(4)-R1 but not for LTB(4)-R2. No apparent affinity binding for the receptors was detected for the CysLT1-selective antagonists montelukast and zafirlukast. LTB(4) functionally mobilized intracellular calcium and inhibited forskolin-stimulated cAMP production in 293 cells. The discovery of this new receptor should aid in further understanding the roles of LTB(4) in pathologies in these tissues and may provide a tool in identification of specific antagonists/agonists for potential therapeutic treatments. PMID- 11006271 TI - Mechanism of phosphorylation of protein kinase B/Akt by a constitutively active 3 phosphoinositide-dependent protein kinase-1. AB - Phosphorylation of Thr(308) in the activation loop and Ser(473) at the carboxyl terminus is essential for protein kinase B (PKB/Akt) activation. However, the biochemical mechanism of the phosphorylation remains to be characterized. Here we show that expression of a constitutively active mutant of mouse 3 phosphoinositide-dependent protein kinase-1 (PDK1(A280V)) in Chinese hamster ovary cells overexpressing the insulin receptor was sufficient to induce PKB phosphorylation at Thr(308) to approximately the same extent as insulin stimulation. Phosphorylation of PKB by PDK1(A280V) was not affected by treatment of cells with inhibitors of phosphatidylinositol 3-kinase or by deletion of the pleckstrin homology (PH) domain of PKB. C(2)-ceramide, a cell-permeable, indirect inhibitor of PKB phosphorylation, did not inhibit PDK1(A280V)-catalyzed PKB phosphorylation in cells and had no effect on PDK1 activity in vitro. On the other hand, co-expression of full-length protein kinase C-related kinase-1 (PRK1/PKN) or 2 (PRK2) inhibited PDK1(A280V)-mediated PKB phosphorylation. Replacing alanine at position 280 with valine or deletion of the PH domain enhanced PDK1 autophosphorylation in vitro. However, deletion of the PH domain of PDK1(A280V) significantly reduced PDK1(A280V)-mediated phosphorylation of PKB in cells. In resting cells, PDK1(A280V) localized in the cytosol and at the plasma membrane. However, PDK1(A280V) lacking the PH domain localized predominantly in the cytosol. Taken together, our findings suggest that the wild-type PDK1 may not be constitutively active in cells. In addition, activation of PDK1 is sufficient to phosphorylate PKB at Thr(308) in the cytosol. Furthermore, the PH domain of PDK1 may play both positive and negative roles in regulating the in vivo function of the enzyme. Finally, unlike the carboxyl-terminal fragment of PRK2, which has been shown to bind PDK1 and allow the enzyme to phosphorylate PKB at both Thr(308) and Ser(473), full-length PRK2 and its related kinase PRK1/PKN may both play negative roles in PKB-mediated downstream biological events. PMID- 11006273 TI - E1A12S-mediated activation of the adenovirus type 12 E2 promoter depends on the histone acetyltransferase activity of p300/CBP. AB - Activation of the transcription unit early region 2 (E2) promoter of the oncogenic adenovirus serotype 12 (Ad12), which regulates the expression of proteins essential for viral replication, requires the assembly of a ternary complex consisting of cAMP response element-binding protein (CREB)-1/activating transcription factor (ATF)-1, the Ad12 12S oncogene product of early region 1A (E1A(12S)), and the co-activator p300/CBP on the E2(Ad12) cAMP response element (E2-CRE). Here we show that the active E2(Ad12) promoter is associated with acetylated histone H4 whereas an E2-CRE point-mutated promoter which is transcriptionally inactive due to its inability to assemble this ternary complex is not bound by acetylated histone H4. The histone deacetylase 1 as well as Roscovitine, which blocks the activation of the histone acetyltransferase (HAT) activity of CBP by cyclin E-Cdk2, prevents E2(Ad12) promoter activation through E1A(12S). p300/CBP counteracts the repressive function of histone deacetylase 1 in a HAT domain-dependent manner whereas the p300/CBP-associated factor PCAF failed to rescue E2(Ad12) promoter activity. E1A(12S) bound p300/CBP displays strong HAT activity. Most interestingly, E1A(12S)-mediated activation of the E2(Ad12) promoter correlates well with the ability of the viral protein to associate with the HAT activity of p300/CBP in vivo. Taken together these data indicate that the recruitment of the HAT activity of p300/CBP by E1A(12S) plays an important role in E2(Ad12) promoter activation. PMID- 11006274 TI - A predicted alpha -helix mediates targeting of the proprotein convertase PC1 to the regulated secretory pathway. AB - The proprotein convertase PC1 is a protease whose activity is largely confined to the dense core secretory granules of neuroendocrine cells. Efficient processing of PC1 substrates in granules requires a mechanism that will both limit the activity of the enzyme to these organelles and promote its targeting to the nascent secretory granules. In the current study, we provide evidence that targeting of PC1 to secretory granules is mediated by alpha-helical structures in its C-terminal tail and, at least in part, is dependent on interactions with specific components of the secretory granule membrane. PMID- 11006275 TI - Receptor-interacting protein 140 directly recruits histone deacetylases for gene silencing. AB - Receptor-interacting protein 140 (RIP140) encodes a histone deacetylase (HDAC) inhibitor-sensitive repressive activity. Direct interaction of RIP140 with HDAC1 and HDAC3 occurs in vitro and in vivo as demonstrated in co-immunoprecipitation and glutathione S-transferase pull-down experiments. The HDAC-interacting domain of RIP140 is mapped to its N-terminal domain, between amino acids 78 and 303 based upon glutathione S-transferase pull-down experiments. In chromatin immunoprecipitation assays, it is demonstrated that histone deacetylation occurs at the chromatin region of the Gal4 binding sites as a result of Gal4 DNA binding domain-tethered RIP expression. The immunocomplexes of RIP140 from cells transfected with RIP140 and HDAC are able to deacetylate histone proteins in vitro. This study presents the first evidence for RIP140 as a negative coregulator for nuclear receptor actions by directly recruiting histone deacetylases and categorizes RIP140 as a novel negative coregulator that is able to directly interact with HDACs. PMID- 11006276 TI - Fidelity and processivity of DNA synthesis by DNA polymerase kappa, the product of the human DINB1 gene. AB - Mammalian DNA polymerase kappa (pol kappa), a member of the UmuC/DinB nucleotidyl transferase superfamily, has been implicated in spontaneous mutagenesis. Here we show that human pol kappa copies undamaged DNA with average single-base substitution and deletion error rates of 7 x 10(-3) and 2 x 10(-3), respectively. These error rates are high when compared to those of most other DNA polymerases. pol kappa also has unusual error specificity, producing a high proportion of T.CMP mispairs and deleting and adding non-reiterated nucleotides at extraordinary rates. Unlike other members of the UmuC/DinB family, pol kappa can processively synthesize chains of 25 or more nucleotides. This moderate processivity may reflect a contribution of C-terminal residues, which include two zinc clusters. The very low fidelity and moderate processivity of pol kappa is novel in comparison to any previously studied DNA polymerase, and is consistent with a role in spontaneous mutagenesis. PMID- 11006277 TI - Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G. AB - The novel pore-forming toxin hemolysin E (HlyE, ClyA, or SheA) consists of a long four-helix bundle with a subdomain (beta tongue) that interacts with target membranes at one pole and an additional helix (alpha(G)) that, with the four long helices, forms a five-helix bundle (tail domain) at the other pole. Random amino acid substitutions that impair hemolytic activity were clustered mostly, but not exclusively, within the tail domain, specifically amino acids within, adjacent to, or interacting with alpha(G). Deletion of amino acids downstream of alpha(G) did not affect activity, but deletions encompassing alpha(G) yielded insoluble and inactive proteins. In the periplasm Cys-285 (alpha(G)) is linked to Cys-87 (alpha(B)) of the four-helix bundle via an intramolecular disulfide. Oxidized HlyE did not form spontaneously in vitro but could be generated by addition of Cu(II) or mimicked by treatment with Hg(II) salts to yield inactive proteins. Such treatments did not affect binding to target membranes nor assembly into non covalently linked octameric complexes once associated with a membrane. However, gel filtration analyses suggested that immobilizing alpha(G) inhibits oligomerization in solution. Thus once associated with a membrane, immobilizing alpha(G) inhibits HlyE activity at a late stage of pore formation, whereas in solution it prevents aggregation and consequent inactivation. PMID- 11006278 TI - Arachidonic acid and nonsteroidal anti-inflammatory drugs induce conformational changes in the human prostaglandin endoperoxide H2 synthase-2 (cyclooxygenase-2). AB - By using the technique of site-directed spin labeling combined with EPR spectroscopy, we have observed that binding of arachidonic acid and nonsteroidal anti-inflammatory drugs induces conformational changes in the human prostaglandin endoperoxide H(2) synthase enzyme (PGHS-2). Line shape broadening resulting from spin-spin coupling of nitroxide pairs introduced into the membrane-binding helices of PGHS-2 was used to calculate the inter-helical distances and changes in these distances that occur in response to binding various ligands. The inter residue distances determined for the PGHS-2 holoenzyme using EPR were 1-7.9 A shorter than those of the crystal structure of the PGHS-2 holoenzyme. However, inter-helical distances calculated and determined by EPR for PGHS-2 complexed with arachidonic acid, flurbiprofen, and SC-58125 were in close agreement with those obtained from the cognate crystal structures. These results indicate that the structure of the solubilized PGHS-2 holoenzyme measured in solution differs from the crystal structure of PGHS-2 holoenzyme obtained by x-ray analysis. Furthermore, binding of ligands induces a conformational change in the holo-PGHS 2, converting it to a structure similar to those obtained by x-ray analysis. Proteolysis protection assays had previously provided circumstantial evidence that binding of heme and non-steroidal anti-inflammatory drugs alters the conformation of PGHS, but the present experiments are the first to directly measure such changes. The finding that arachidonate can also induce a conformational change in PGHS-2 was unexpected, and the magnitude of changes suggests this structural flexibility may be integral to the cyclooxygenase catalytic mechanism. PMID- 11006279 TI - Identification of the substrate interaction site in the N-terminal membrane anchor segment of thromboxane A2 synthase by determination of its substrate analog conformational changes using high resolution NMR technique. AB - The present studies describe an investigation for the interaction of N-terminal membrane anchor domain of thromboxane A(2) synthase (TXAS) with its substrate analog in a membrane-bound environment using the two-dimensional NMR technique. TXAS and prostaglandin I(2) synthase (PGIS), respectively, convert the same substrate, prostaglandin H(2) (PGH(2)), to thromboxane A(2) and prostaglandin I(2), which have opposite biological functions. Our topology studies have indicated that the N-terminal region of TXAS has a longer N-terminal endoplasmic reticulum (ER) membrane anchor region compared with the same segment proposed for PGIS. The differences in their interaction with the ER membrane may have an important impact to facilitate their common substrate, PGH(2), across the membrane into their active sites from the luminal to the cytoplasmic side of the ER. To test this hypothesis, we first investigated the interaction of the TXAS N terminal membrane anchor domain with its substrate analog. A synthetic peptide corresponding to the N-terminal membrane anchor domain (residues 1-35) of TXAS, which adopted a stable helical structure and exhibited a membrane anchor function in the membrane-bound environment, was used to interact with a stable PGH(2) analog,. High resolution two-dimensional NMR experiments, NOESY and TOCSY, were performed to solve the solution structures of in a membrane-mimicking environment using dodecylphosphocholine micelles. Different conformations were clearly observed in the presence and absence of the TXAS N-terminal membrane anchor domain. Through combination of the two-dimensional NMR experiments, completed (1)H NMR assignments of were obtained, and the data were used to construct three dimensional structures of in H(2)O and dodecylphosphocholine micelles, showing the detailed conformation change upon the interaction with the membrane anchor domain. The observation supported the presence of a substrate interaction site in the N-terminal region. The combination of the structural information of and was able to simulate a solution structure of the unstable TXAS and PGIS substrate, PGH(2). PMID- 11006280 TI - Spatial control of Ca2+ signaling by nicotinic acid adenine dinucleotide phosphate diffusion and gradients. AB - Intracellular Ca(2+) is able to control numerous cellular responses through complex spatiotemporal organization. Ca(2+) waves mediated by inositol trisphosphate or ryanodine receptors propagate by Ca(2+)-induced Ca(2+) release and therefore do not have an absolute requirement for a gradient in either inositol trisphosphate or cyclic ADP-ribose, respectively. In contrast, we report that although Ca(2+) increases induced by nicotinic acid adenine dinucleotide phosphate (NAADP) are amplified by Ca(2+)-induced Ca(2+) release locally, Ca(2+) waves mediated by NAADP have an absolute requirement for an NAADP gradient. If NAADP is increased such that its concentration is spatially uniform in one region of an egg, the Ca(2+) increase occurs simultaneously throughout this area, and only where there is diffusion out of this area to establish an NAADP gradient is there a Ca(2+) wave. A local increase in NAADP results in a Ca(2+) increase that spreads by NAADP diffusion. NAADP diffusion is restricted at low but not high concentrations of NAADP, indicating that NAADP diffusion is strongly influenced by binding to immobile and saturable sites, probably the NAADP receptor itself. Thus, the range of action of NAADP can be tuned by its concentration from that of a local messenger, like Ca(2+), to that of a global messenger, like IP(3) or cyclic ADP-ribose. PMID- 11006281 TI - Roles of two homotetrameric kinesins in sea urchin embryonic cell division. AB - To improve our understanding of the roles of microtubule cross-linking motors in mitosis, we analyzed two sea urchin embryonic kinesin-related proteins. It is striking to note that both of these proteins behave as homotetramers, but one behaves as a more compact molecule than the other. These observations suggest that these two presumptive motors could cross-link microtubules into bundles with different spacing. Both motors localize to mitotic spindles, and antibody microinjection experiments suggest that they have mitotic functions. Thus, one of these kinesin-related proteins may cross-link spindle microtubules into loose bundles that are "tightened" by the other. PMID- 11006282 TI - Ribonucleotide reductase, a possible agent in deoxyribonucleotide pool asymmetries induced by hypoxia. AB - While investigating the basis for marked natural asymmetries in deoxyribonucleoside triphosphate (dNTP) pools in mammalian cells, we observed that culturing V79 hamster lung cells in a 2% oxygen atmosphere causes 2-3-fold expansions of the dATP, dGTP, and dTTP pools, whereas dCTP declines by a comparable amount. Others have made similar observations and have proposed that, because O(2) is required for formation of the catalytically essential oxygen bridged iron center in ribonucleotide reductase, dCTP depletion at low oxygen tension results from direct or indirect effects upon ribonucleotide reductase. We have tested the hypothesis that oxygen limitation affects ribonucleotide specificity using recombinant mouse ribonucleotide reductase and an assay that permits simultaneous monitoring of the reduction of all four nucleotide substrates. Preincubation and assay of the enzyme in an anaerobic chamber caused only partial activity loss. Accordingly, we treated the enzyme with hydroxyurea, followed by removal of the hydroxyurea and exposure to atmospheres of varying oxygen content. The activity was totally depleted by hydroxyurea treatment and nearly fully regained by exposure to air. By the criterion of activities regained at different oxygen tensions, we found CDP reduction not to be specifically sensitive to oxygen depletion; however, GDP reduction was specifically sensitive. The basis for the differential response to reactivation by O(2) is not known, but it evidently does not involve varying rates of reactivation of different allosteric forms of the enzyme or altered response to allosteric effectors at reduced oxygen tension. PMID- 11006283 TI - The AROM gene, spliced mRNAs encoding new DNA/RNA-binding proteins are transcribed from the opposite strand of the melanin-concentrating hormone gene in mammals. AB - Melanin-concentrating hormone (MCH) mRNA expression is induced by nerve growth factor and lithium in PC12 cells, whereas three large MCH RNA species are found in untreated cells. In this study, we investigated the structures, regulations of expression, and putative functions of these transcripts. Northern blot, rapid amplification of cDNA ends-polymerase chain reaction, reverse transcriptase polymerase chain reaction, and sequencing experiments demonstrated that they are antisense RNAs complementary to the MCH gene. Two classes of antisense RNAs could be discriminated as follows: 1) non-coding unspliced RNAs that overlap mainly the coding part of the MCH gene; 2) spliced variant mRNAs complementary to the 3' flanking end of the MCH gene and that encode putative proteins containing DNA/RNA binding domains. We named this new transcriptional unit AROM for antisense-RNA overlapping-MCH gene. Spliced variant AROM mRNAs are expressed in a broad range of rat organs. Western blot and immunohistochemistry experiments revealed several proteins with cytoplasmic but also nuclear localization in PC12 cells. Time course studies during nerve growth factor and lithium treatment of PC12 cells indicated a reciprocal regulation of the MCH and AROM gene transcripts, reflected also at the level of AROM proteins. The major translational product is a 64-kDa protein (AROM-p64). Recombinant AROM-p64 displayed high binding to single stranded DNA and poly(A) homopolymers suggesting that this protein could play a role in mRNA maturation/metabolism. PMID- 11006284 TI - The protein-tyrosine kinase fer associates with signaling complexes containing insulin receptor substrate-1 and phosphatidylinositol 3-kinase. AB - In a screen for 3T3-F442A adipocyte proteins that bind SH2 domains, we isolated a cDNA encoding Fer, a nonreceptor protein-tyrosine kinase of the Fes/Fps family that contains a functional SH2 domain. A truncated splicing variant, iFer, was also cloned. iFer is devoid of both the tyrosine kinase domain and a functional SH2 domain but displays a unique 42-residue C terminus and retains the ability to form oligomers with Fer. Expression of both Fer and iFer proteins are strikingly increased upon differentiation of 3T3-L1 fibroblasts to adipocytes. Platelet derived growth factor treatment of the cultured adipocytes caused rapid tyrosine phosphorylation of Fer and its recruitment to complexes containing platelet derived growth factor receptor and the p85 regulatory subunit of phosphatidylinositol (PI) 3-kinase. Insulin treatment of 3T3-L1 adipocytes stimulated association of Fer with complexes containing tyrosine phosphorylated IRS-1 and PI 3-kinase but did not stimulate tyrosine phosphorylation of Fer. PI 3 kinase activity in anti-Fer immunoprecipitates was also acutely activated by insulin treatment of cultured adipocytes. These data demonstrate the presence of Fer tyrosine kinase in insulin signaling complexes, suggesting a role of Fer in insulin action. PMID- 11006285 TI - Modeling the late steps in HIV-1 retroviral integrase-catalyzed DNA integration. AB - Model oligodeoxyribonucleotide substrates representing viral DNA integration intermediates with a gap and a two-nucleotide 5' overhang were used to examine late steps in human immunodeficiency virus, type 1 (HIV-1) retroviral integrase (IN)-catalyzed DNA integration in vitro. HIV-1 or avian myeloblastosis virus reverse transcriptase (RT) were capable of quantitatively filling in the gap to create a nicked substrate but did not remove the 5' overhang. HIV-1 IN also failed to remove the 5' overhang with the gapped substrate. However, with a nicked substrate formed by RT, HIV-1 IN removed the overhang and covalently closed the nick in a disintegration-like reaction. The efficiency of this closure reaction was very low. Such closure was not stimulated by the addition of HMG (I/Y), suggesting that this protein only acts during the early processing and joining reactions. Addition of Flap endonuclease-1, a nuclease known to remove 5' overhangs, abolished the closure reaction catalyzed by IN. A series of base pair inversions, introduced into the HIV-1 U5 long terminal repeat sequence adjacent to and/or including the conserved CA dinucleotide, produced no or only a small decrease in the HIV-1 IN-dependent strand closure reaction. These same mutations caused a significant decrease in the efficiency of concerted DNA integration by a modified donor DNA in vitro, suggesting that recognition of the ends of the long terminal repeat sequence is required only in the early steps of DNA integration. Finally, a combination of HIV-1 RT, Flap endonuclease-1, and DNA ligase is capable of quantitatively forming covalently closed DNA with these model substrates. These results support the hypothesis that cellular enzyme(s) may catalyze the late steps of retroviral DNA integration. PMID- 11006286 TI - Differential targeting of beta -adrenergic receptor subtypes and adenylyl cyclase to cardiomyocyte caveolae. A mechanism to functionally regulate the cAMP signaling pathway. AB - Differential modes for beta(1)- and beta(2)-adrenergic receptor (AR) regulation of adenylyl cyclase in cardiomyocytes is most consistent with spatial regulation in microdomains of the plasma membrane. This study examines whether caveolae represent specialized subdomains that concentrate and organize these moieties in cardiomyocytes. Caveolae from quiescent rat ventricular cardiomyocytes are highly enriched in beta(2)-ARs, Galpha(i), protein kinase A RIIalpha subunits, caveolin 3, and flotillins (caveolin functional homologues); beta(1)-ARs, m(2)-muscarinic cholinergic receptors, Galpha(s), and cardiac types V/VI adenylyl cyclase distribute between caveolae and other cell fractions, whereas protein kinase A RIalpha subunits, G protein-coupled receptor kinase-2, and clathrin are largely excluded from caveolae. Cell surface beta(2)-ARs localize to caveolae in cardiomyocytes and cardiac fibroblasts (with markedly different beta(2)-AR expression levels), indicating that the fidelity of beta(2)-AR targeting to caveolae is maintained over a physiologic range of beta(2)-AR expression. In cardiomyocytes, agonist stimulation leads to a marked decline in the abundance of beta(2)-ARs (but not beta(1)-ARs) in caveolae. Other studies show co immunoprecipitation of cardiomyocytes adenylyl cyclase V/VI and caveolin-3, suggesting their in vivo association. However, caveolin is not required for adenylyl cyclase targeting to low density membranes, since adenylyl cyclase targets to low buoyant density membrane fractions of HEK cells that lack prototypical caveolins. Nevertheless, cholesterol depletion with cyclodextrin augments agonist-stimulated cAMP accumulation, indicating that caveolae function as negative regulators of cAMP accumulation. The inhibitory interaction between caveolae and the cAMP signaling pathway as well as domain-specific differences in the stoichiometry of individual elements in the beta-AR signaling cascade represent important modifiers of cAMP-dependent signaling in the heart. PMID- 11006287 TI - A single regulatory module of the carbamoylphosphate synthetase I gene executes its hepatic program of expression. AB - A 469-base pair (bp) upstream regulatory fragment (URF) and the proximal promoter of the carbamoylphosphate synthetase I (CPS) gene were analyzed for their role in the regulation of spatial, developmental, and hormone-induced expression in vivo. The URF is essential and sufficient for hepatocyte-specific expression, periportal localization, perinatal activation and induction by glucocorticoids, and cAMP in transgenic mice. Before birth, the transgene is silent but can be induced by cAMP and glucocorticoids, indicating that these compounds are responsible for the activation of expression at birth. A 102-bp glucocorticoid response unit within the URF, containing binding sites for HNF3, C/EBP, and the glucocorticoid receptor, is the main determinant of the hepatocyte-specific and hormone-controlled activity. Additional sequences are required for a productive interaction between this minimal response unit and the core CPS promoter. These results show that the 469-bp URF, and probably only the 102-bp glucocorticoid response unit, functions as a regulatory module, in that it autonomously executes a correct spatial, developmental and hormonal program of CPS expression in the liver. PMID- 11006288 TI - TATA-binding protein and the Gal4 transactivator do not bind to promoters cooperatively. AB - The yeast Gal4 protein, like many activators, binds TATA-binding protein (TBP) directly in vitro. It has been speculated that this protein-protein interaction is important for Gal4p-mediated activation of transcription, but little work has been done to test specific models involving this interaction. In this study, the effect of Gal4p on TBP-TATA binding is addressed. Specifically, it is asked if the Gal4p-TBP interaction can support cooperative binding of the two factors to promoters. It is easy to see how such an event could stimulate transcription, particularly from promoters with a non-consensus TATA box. In vitro, however, a derivative of Gal4p (Gal4-(1-93+768-881)) containing the DNA-binding, dimerization, and activation domains does not bind to promoter DNA cooperatively with either recombinant, purified TBP, or with protein from a yeast crude extract. In vivo, reporter gene experiments using promoters with differing TBP affinities reveal no major Gal4p-mediated stimulation of TBP function from weak TATA boxes, as would be predicted if the proteins bind cooperatively. Furthermore, native Gal4p and a potent Gal4p-based artificial activator lacking a TBP-binding activation domain support similar ratios of transcription from a series of promoters identical except for mutations in the TATA box. It is concluded that Gal4p and TBP do not bind cooperatively to promoters and that this mechanism does not contribute substantially to Gal4p-mediated transcriptional activation. PMID- 11006289 TI - Active DNA topoisomerase IIalpha is a component of the salt-stable centrosome core. AB - Recently, we reported that the monoclonal antibody specific for human DNA topoisomerase IIalpha, Ki-S1, stains not only the nuclei of human A431 cells but also extranuclear structures suggestive of centrosomes (Meyer, K. N., Kjeldsen, E., Straub, T., Knudsen, B. K., Kikuchi, A., Hickson, I. D., Kreipe, H., and Boege, F. (1997) J. Cell Biol. 136, 775-788). Here, we confirm colocalization of Ki-S1 with the centrosomal marker gamma-tubulin. In addition, we show labeling of centrosomes by peptide antibodies against the N and C termini of human topoisomerase IIalpha. Probing Western blots of isolated centrosomes with topoisomerase IIalpha antibodies, we demonstrate a protein band of 170 kDa. Moreover, isolated centrosomes exhibited DNA decatenation and relaxation activity correlated to the amount of topoisomerase IIalpha protein in the same way as seen in the pure recombinant enzyme. Topoisomerase IIalpha epitopes could not be removed from centrosomes by salt extraction, DNase treatment, or RNase treatment, procedures that completely removed the enzyme from nuclei. Taken together, these observations suggest that active topoisomerase IIalpha is bound tightly to the centrosome in a DNA-independent manner. Because such centrosomal topoisomerase IIalpha was also present in quiescent lymphocytes devoid of topoisomerase IIalpha in the nuclei, we assume that it might be a long-lived storage form. PMID- 11006290 TI - Age-related macular degeneration. The lipofusion component N-retinyl-N retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells. AB - 10-20% of individuals over the age of 65 suffer from age-related macular degeneration (AMD), the leading cause of severe visual impairment in humans living in developed countries. The pathogenesis of this complex disease is poorly understood, and no efficient therapy or prevention exists to date. A precondition for AMD appears to be the accumulation of the age pigment lipofuscin in lysosomes of retinal pigment epithelial (RPE) cells. In AMD, these cells seem to die by apoptosis with subsequent death of photoreceptor cells, and light may accelerate the disease process. Intracellular factors leading to cell death are not known. Here we show that the lipophilic cation N-retinyl-N-retinylidene ethanolamine (A2E), a lipofuscin component, induces apoptosis in RPE and other cells at concentrations found in human retina. Apoptosis is accompanied by the appearance of the proapoptotic proteins cytochrome c and apoptosis-inducing factor in the cytoplasm and the nucleus. Biochemical examinations show that A2E specifically targets cytochrome oxidase (COX). With both isolated mitochondria and purified COX, A2E inhibits oxygen consumption synergistically with light. Inhibition is reversed by the addition of cytochrome c or cardiolipin, a negatively charged phospholipid that facilitates the binding of cytochrome c to membranes. Succinate dehydrogenase activity is not altered by A2E. We suggest that A2E can act as a proapoptotic molecule via a mitochondria-related mechanism, possibly through site specific targeting of this cation to COX. Loss of RPE cell viability through inhibition of mitochondrial function might constitute a pivotal step toward the progressive degeneration of the central retina. PMID- 11006291 TI - Sequence requirements for protein-primed initiation and elongation of phage O29 DNA replication. AB - The double-stranded linear DNA of Bacillus subtilis phage O29 is replicated by a mechanism in which a terminal protein (TP) acts as a primer. The second 3' terminal nucleotide of the template directs the incorporation of the 5'-terminal nucleotide into the TP, giving rise to the initiation complex TP-dAMP. Elongation then proceeds by a sliding-back mechanism in which the dAMP covalently linked to the TP pairs to the 3'-terminal nucleotide of the template strand to recover full length DNA. We have studied the sequence requirements for efficient initiation of replication using mutated TP-free double-stranded DNA fragments. Efficient initiation only requires the terminal repetition 5'-AA. The 3'-terminal T, although not used as template, increases the affinity of DNA polymerase for the initiator nucleotide; in addition, although to a minor extent, the third 3' terminal position also directs the formation of the initiation complex and modulates the initiation rate at the second position. Efficient elongation requires a previous sliding-back, demanding again a repetition of two nucleotides at the 3' end; if the sliding-back is prevented, a residual elongation can proceed directly from the second position or after jumping back from the third to the first position. PMID- 11006292 TI - Human alpha (1,3)-fucosyltransferase IV (FUTIV) gene expression is regulated by elk-1 in the U937 cell line. AB - The alpha1,3-fucosyltransferase IV (FucTIV) encoded by its gene (FUTIV) is responsible for synthesis of Le(x) (Galbeta4[Fucalpha3]GlcNAcbeta3Galbeta1,R), which causes compaction in the morula stage of the preimplantation mouse embryo, as well as alpha1,3-fucosylation at multiple internal GlcNAc of unbranched poly-N acetyllactosamine, termed "myeloglycan," the physiological epitope of E-selectin. Since myeloglycan-type structure is also expressed in various types of human cancer and may mediate E-selectin-dependent metastasis, expression of FUTIV is oncodevelopmentally regulated. The mechanisms controlling FUTIV expression remain to be clarified. In this report, we further characterize FUTIV gene structure and define a non-TATA box-dependent transcriptional start region just upstream from the translational start. FUTIV promoter/reporter fusion constructs defined a "full-length" promoter and highly active fragments in the macrophage-derived U937 and myeloid HL60 cell lines. One highly active fragment contains a consensus binding site for the Ets-1 transcription factor (Withers, D. A., and Hakomori, S. (1997) Glycoconj. J. 14, 764). Gel shift analysis shows specific binding to this site in nuclear extracts from U937 cells. Mutation of the Ets consensus site significantly reduces FUTIV promoter activity in both cell lines. Gel supershift and dominant negative cotransfection experiments identified the Ets family member Elk-1 as one component binding and regulating the FUTIV promoter in U937 cells. The significance of FUTIV regulation by Elk-1 is discussed. PMID- 11006293 TI - A general approach for identification of RNA-protein cross-linking sites within native human spliceosomal small nuclear ribonucleoproteins (snRNPs). Analysis of RNA-protein contacts in native U1 and U4/U6.U5 snRNPs. AB - We describe a novel approach to identify RNA-protein cross-linking sites within native small nuclear ribonucleoprotein (snRNP) particles from HeLa cells. It combines immunoprecipitation of the UV-irradiated particles under semi-denaturing conditions with primer extension analysis of the cross-linked RNA moiety. In a feasibility study, we initially identified the exact cross-linking sites of the U1 70-kDa (70K) protein in stem-loop I of U1 small nuclear RNA (snRNA) within purified U1 snRNPs and then confirmed the results by a large-scale preparation that allowed N-terminal sequencing and matrix-assisted laser desorption ionization mass spectrometry of purified cross-linked peptide-oligonucleotide complexes. We identified Tyr(112) and Leu(175) within the RNA-binding domain of the U1 70K protein to be cross-linked to G(28) and U(30) in stem-loop I, respectively. We further applied our immunoprecipitation approach to HeLa U5 snRNP, as part of purified 25 S U4/U6.U5 tri-snRNPs. Cross-linking sites between the U5-specific 220-kDa protein (human homologue of Prp8p) and the U5 snRNA were located at multiple nucleotides within the highly conserved loop 1 and at one site in internal loop 1 of U5 snRNA. The cross-linking of four adjacent nucleotides indicates an extended interaction surface between loop 1 and the 220 kDa protein. In summary, our approach provides a rapid method for identification of RNA-protein contact sites within native snRNP particles as well as other ribonucleoprotein particles. PMID- 11006294 TI - Identification of ISC1 (YER019w) as inositol phosphosphingolipid phospholipase C in Saccharomyces cerevisiae. AB - Sphingolipids have emerged as novel bioactive mediators in eukaryotic cells including yeast. It has been proposed that sphingomyelin (SM) hydrolysis and the concomitant generation of ceramide are involved in various stress responses in mammalian cells. The yeast Saccharomyces cerevisiae has inositol phosphosphingolipids (IPS) instead of SM and glycolipids, and synthesis of IPS is indispensable to its growth. Although the genes responsible for the synthesis of IPS have been identified, the gene(s) for the degradation of IPS has not been reported. Here we show that ISC1 (YER019w), which has homology to bacterial neutral sphingomyelinase (SMase), encodes IPS phospholipase C (IPS-PLC). First, we observed that overexpression of ISC1 greatly increased neutral SMase activity, and this activity was dependent on the presence of phosphatidylserine. Cells deleted in ISC1 demonstrated negligible neutral SMase activity. Because yeast cells have IPS instead of SM, we investigated whether IPS are the physiologic substrates of this enzyme. Lysates of ISC1-overexpressing cells demonstrated very high PLC activities on IPS. Deletion of ISC1 eliminated endogenous IPS-PLC activities. Labeling yeast cells with [(3)H]dihydrosphingosine showed that IPS were increased in the deletion mutant cells. This study identifies the first enzyme involved in catabolism of complex sphingolipids in S. cerevisiae. PMID- 11006295 TI - Compromised energetics in the adenylate kinase AK1 gene knockout heart under metabolic stress. AB - Rapid exchange of high energy carrying molecules between intracellular compartments is essential in sustaining cellular energetic homeostasis. Adenylate kinase (AK)-catalyzed transfer of adenine nucleotide beta- and gamma-phosphoryls has been implicated in intracellular energy communication and nucleotide metabolism. To demonstrate the significance of this reaction in cardiac energetics, phosphotransfer dynamics were determined by [(18)O]phosphoryl oxygen analysis using( 31)P NMR and mass spectrometry. In hearts with a null mutation of the AK1 gene, which encodes the major AK isoform, total AK activity and beta phosphoryl transfer was reduced by 94% and 36%, respectively. This was associated with up-regulation of phosphoryl flux through remaining minor AK isoforms and the glycolytic phosphotransfer enzyme, 3-phosphoglycerate kinase. In the absence of metabolic stress, deletion of AK1 did not translate into gross abnormalities in nucleotide levels, gamma-ATP turnover rate or creatine kinase-catalyzed phosphotransfer. However, under hypoxia AK1-deficient hearts, compared with the wild type, had a blunted AK-catalyzed phosphotransfer response, lowered intracellular ATP levels, increased P(i)/ATP ratio, and suppressed generation of adenosine. Thus, although lack of AK1 phosphotransfer can be compensated in the absence of metabolic challenge, under hypoxia AK1-knockout hearts display compromised energetics and impaired cardioprotective signaling. This study, therefore, provides first direct evidence that AK1 is essential in maintaining myocardial energetic homeostasis, in particular under metabolic stress. PMID- 11006296 TI - Novel catalytic mechanism of nucleophilic substitution by asparagine residue involving cyanoalanine intermediate revealed by mass spectrometric monitoring of an enzyme reaction. AB - l-2-Haloacid dehalogenase from Pseudomonas sp. YL catalyzes the hydrolytic dehalogenation, in which Asp(10) acts as a nucleophile to attack the alpha-carbon of l-2-haloalkanoates to form an ester intermediate, which is subsequently hydrolyzed to produce d-2-hydroxyalkanoates. Surprisingly, replacement of the catalytic residue, Asp(10), by Asn did not result in total inactivation of the enzyme (Kurihara, T., Liu, J.-Q., Nardi-Dei, V., Koshikawa, H., Esaki, N., and Soda, K. (1995) J. Biochem. 117, 1317-1322). In this study, we monitored the D10N mutant enzyme reaction by ion-spray mass spectrometry, and found that the enzyme shows a unique structural change when it was incubated with the substrate, l-2 chloropropionate. LC/MS and tandem MS/MS analyses revealed that Asn(10) attacks the substrate to form an imidate, and a proton and d-lactic acid are eliminated to produce a nitrile (beta-cyanoalanine residue), followed by hydrolysis to reproduce Asn(10). This is the first report of the function of Asn to catalyze nucleophilic substitution through its conversion to beta-cyanoalanine residue as an intermediate structure. Also, these results demonstrate that mass spectrometry is remarkably useful in monitoring enzyme reactions. PMID- 11006297 TI - Regulation of protein kinase C by the cytoskeletal protein calponin. AB - Previous studies from this laboratory have shown that, upon agonist activation, calponin co-immunoprecipitates and co-localizes with protein kinase Cepsilon (PKCepsilon) in vascular smooth muscle cells. In the present study we demonstrate that calponin binds directly to the regulatory domain of PKC both in overlay assays and, under native conditions, by sedimentation with lipid vesicles. Calponin was found to bind to the C2 region of both PKCepsilon and PKCalpha with possible involvement of C1B. The C2 region of PKCepsilon binds to the calponin repeats with a requirement for the region between amino acids 160 and 182. We have also found that calponin can directly activate PKC autophosphorylation. By using anti-phosphoantibodies to residue Ser-660 of PKCbetaII, we found that calponin, in a lipid-independent manner, increased auto-phosphorylation of PKCalpha, -epsilon, and -betaII severalfold compared with control conditions. Similarly, calponin was found to increase the amount of (32)P-labeled phosphate incorporated into PKC from [gamma-(32)P]ATP. We also observed that calponin addition strongly increased the incorporation of radiolabeled phosphate into an exogenous PKC peptide substrate, suggesting an activation of enzyme activity. Thus, these results raise the possibility that calponin may function in smooth muscle to regulate PKC activity by facilitating the phosphorylation of PKC. PMID- 11006298 TI - Mechanism of chalcone synthase. pKa of the catalytic cysteine and the role of the conserved histidine in a plant polyketide synthase. AB - Polyketide synthases (PKS) assemble structurally diverse natural products using a common mechanistic strategy that relies on a cysteine residue to anchor the polyketide during a series of decarboxylative condensation reactions that build the final reaction product. Crystallographic and functional studies of chalcone synthase (CHS), a plant-specific PKS, indicate that a cysteine-histidine pair (Cys(164)-His(303)) forms part of the catalytic machinery. Thiol-specific inactivation and the pH dependence of the malonyl-CoA decarboxylation reaction were used to evaluate the potential interaction between these two residues. Inactivation of CHS by iodoacetamide and iodoacetic acid targets Cys(164) in a pH dependent manner (pK(a) = 5.50). The acidic pK(a) of Cys(164) suggests that an ionic interaction with His(303) stabilizes the thiolate anion. Consistent with this assertion, substitution of a glutamine for His(303) maintains catalytic activity but shifts the pK(a) of the thiol to 6.61. Although the H303A mutant was catalytically inactive, the pH-dependent incorporation of [(14)C]iodoacetamide into this mutant exhibits a pK(a) = 7.62. Subsequent analysis of the pH dependence of the malonyl-CoA decarboxylation reaction catalyzed by wild-type CHS and the H303Q and C164A mutants also supports the presence of an ion pair at the CHS active site. Structural and sequence conservation of a cysteine-histidine pair in the active sites of other PKS implies that a thiolate-imidazolium ion pair plays a central role in polyketide biosynthesis. PMID- 11006299 TI - Neuronal Ca2+ sensor-1/frequenin functions in an autocrine pathway regulating Ca2+ channels in bovine adrenal chromaffin cells. AB - NCS-1/frequenin belongs to a family of EF-hand-containing Ca(2+) sensors expressed mainly in neurons. Overexpression of NCS-1/frequenin has been shown to stimulate neurotransmitter release but little else is known of its cellular roles. We have constructed an EF-hand mutant, NCS-1(E120Q), as a likely dominant inhibitor of cellular NCS-1 function. Recombinant NCS-1(E120Q) showed an impaired Ca(2+)-dependent conformational change but could still bind to cellular proteins. Transient expression of this mutant, but not NCS-1, in bovine adrenal chromaffin cells increased non-L-type Ca(2+) channel currents. Cells expressing NCS-1(E120Q) no longer responded effectively to the removal of autocrine purinergic/opioid inhibition of Ca(2+) currents but still showed voltage-dependent facilitation. These data are consistent with the existence of both voltage-dependent and voltage-independent pathways for Ca(2+) channel inhibition in chromaffin cells. Our results suggest a novel function for NCS-1 specific for the voltage independent autocrine pathway that negatively regulates non-L-type Ca(2+) channels in chromaffin cells. PMID- 11006300 TI - Turgor, temperature and the growth of plant cells: using Chara corallina as a model system. AB - Rapid changes in turgor pressure (P:) and temperature (T:) are giving new information about the mechanisms of plant growth. In the present work, single internode cells of the large-celled alga Chara corallina were used as a model for plant growth. P was changed without altering the chemical environment of the wall while observing growth without elastic changes. When P: was measured before any changes, the original growth rate bore no relationship to the original P. However, if P of growing cells was decreased, growth responded immediately without evidence for rapid changes in wall physical properties. Growth occurred only above a 0.3 MPa threshold, and increasing P caused small increases in growth that became progressively larger as P rose, resulting in a curvilinear response overall. The small changes in growth close to the threshold may explain early failures to detect these responses. When T was lowered, the elastic properties of the cell were unaffected, but growth was immediately inhibited. The lower T caused P to decrease, but returning P to its original value did not return growth to its original rate. The decreased P at low T occurred because of T effects on the osmotic potential of the cell. At above-normal P, growth partially resumed at low T Therefore, growth required a P-sensitive process that was also T-sensitive. Because elastic properties were little affected by T, but growth was markedly affected, the process is likely to involve metabolism. The rapidity of its response to P and T probably excludes the participation of changes in gene expression. PMID- 11006301 TI - Water relations and leaf expansion: importance of time scale. AB - The role of leaf water relations in controlling cell expansion in leaves of water stressed maize and barley depends on time scale. Sudden changes in leaf water status, induced by sudden changes in humidity, light and soil salinity, greatly affect leaf elongation rate, but often only transiently. With sufficiently large changes in salinity, leaf elongation rates are persistently reduced. When plants are kept fully turgid throughout such sudden environmental changes, by placing their roots in a pressure chamber and raising the pressure so that the leaf xylem sap is maintained at atmospheric pressure, both the transient and persistent changes in leaf elongation rate disappear. All these responses show that water relations are responsible for the sudden changes in leaf elongation rate resulting from sudden changes in water stress and putative root signals play no part. However, at a time scale of days, pressurization fails to maintain high rates of leaf elongation of plants in either saline or drying soil, indicating that root signals are overriding water relations effects. In both saline and drying soil, pressurization does raise the growth rate during the light period, but a subsequent decrease during the dark results in no net effect on leaf growth over a 24 h period. When transpirational demand is very high, however, growth promoting effects of pressurization during the light period outweigh any reductions in the dark, resulting in a net increase in growth of pressurized plants over 24 h. Thus leaf water status can limit leaf expansion rates during periods of high transpiration despite the control exercised by hormonal effects on a 24 h basis. PMID- 11006302 TI - Spatial distributions of expansion rate, cell division rate and cell size in maize leaves: a synthesis of the effects of soil water status, evaporative demand and temperature. AB - The spatial distributions of leaf expansion rate, cell division rate and cell size was examined under contrasting soil water conditions, evaporative demands and temperatures in a series of experiments carried out in either constant or naturally fluctuating conditions. They were examined in the epidermis and all leaf tissues. (1) Meristem temperature affected relative elongation rate by a constant ratio at all positions in the leaf. If expressed per unit thermal time, the distribution of relative expansion rate was independent of temperature and was similar in all experiments with low evaporative demand and no water deficit. This provides a reference distribution, characteristic of the studied genotype, to which any distribution in stressed plants can be compared. (2) Evaporative demand and soil water deficit affected independently the distribution of relative elongation rate and had near-additive effects. For a given stress, a nearly constant difference was observed, at all positions of the leaf, between the relative elongation rates of stressed plants and those of control plants. This caused a reduction in the length of the zone with tissue elongation. (3) Methods for calculating cell division rate in the epidermis and in all leaf tissues are proposed and discussed. In control plants, the zone with cell division was 30 mm and 60 mm long in the epidermis and in whole tissues, respectively. Both this length and relative division rate were reduced by soil water deficit. The size of epidermal and of mesophyll cells was nearly unaffected in the leaf zone with both cell division and tissue expansion, suggesting that water deficit affects tissue expansion rate and cell division rate to the same extent. Conversely, cell size of epidermis and mesophyll were reduced by water deficit in mature parts of the leaf. PMID- 11006303 TI - Leaf development in Ricinus communis during drought stress: dynamics of growth processes, of cellular structure and of sink-source transition. AB - Dicot leaf growth is characterized by partly transient tip-to-base gradients of growth processes, structure and function. These gradients develop dynamically and interact with dynamically developing stress conditions like drought. In Ricinus communis plants growing under well-watered and drought conditions growth rates peaked during the late night and minimal values occurred in the late afternoon. During this diurnal course the leaf base always showed much higher rates than the leaf tip. The amplitude of this diurnal course decreased when leaves approached maturity and during drought stress without any significant alteration of the diurnal pattern and it increased during the first days after rewatering. Unique relationships between leaf size and cytological structure were observed. This provided the framework for the analysis of changes in assimilation, transpiration and dark respiration, chlorophyll, protein, carbohydrate, and amino acid concentrations, and of activities of sink-source-related enzymes at the leaf tip and base during leaf development in well-watered and drought-stressed plants. Gas exchange was dominated by physiological rather than by anatomical properties (stomatal density). Tip-to-base gradients in carbohydrate concentrations per dry weight and sink-source-related enzymes were absent, whereas significant gradients were found in amino acid concentrations per dry weight. During drought stress, growing leaves developed source function at smaller leaf size, before specific physiological adaptations to drought occurred. The relevance of the developmental status of individual leaves for the drought-stress response and of the structural changes for the biochemical composition changes is discussed. PMID- 11006304 TI - Water uptake by roots: effects of water deficit. AB - The variable hydraulic conductivity of roots (Lp(r)) is explained in terms of a composite transport model. It is shown how the complex, composite anatomical structure of roots results in a composite transport of both water and solutes. In the model, the parallel apoplastic and cell-to-cell (symplastic and transcellular) pathways play an important role as well as the different tissues and structures arranged in series within the root cylinder (epidermis, exodermis, cortex, endodermis, stelar parenchyma). The roles of Casparian bands and suberin lamellae in the root's endo- and exodermis are discussed. Depending on the developmental state of these apoplastic barriers, the overall hydraulic resistance of roots is either more evenly distributed across the root cylinder (young unstressed roots) or is concentrated in certain layers (exo- and endodermis in older stressed roots). The reason for the variability of root Lp(r), is that hydraulic forces cause a dominating apoplastic flow of water around protoplasts, even in the endodermis and exodermis. In the absence of transpiration, water flow is osmotic in nature which causes a high resistance as water passes across many membranes on its passage across the root cylinder. The model allows for a high capability of roots to take up water in the presence of high rates of transpiration (high demands for water from the shoot). By contrast, the hydraulic conductance is low, when transpiration is switched off. Overall, this results in a non-linear relationship between water flow and forces (gradients of hydrostatic and osmotic pressure) which is otherwise hard to explain. The model allows for special root characteristics such as a high hydraulic conductivity (water permeability) in the presence of a low permeability of nutrient ions once taken up into the stele by active processes. Low root reflection coefficients are in line with the idea of some apoplastic bypasses for water within the root cylinder. According to the composite transport model, the switch from the hydraulic to the osmotic mode is purely physical. In the presence of heavily suberized roots, the apoplastic component of water flow may be too small. Under these conditions, a regulation of radial water flow by water channels dominates. Since water channels are under metabolic control, this component represents an 'active' element of regulation. Composite transport allows for an optimization of the water balance of the shoot in addition to the well-known phenomena involved in the regulation of water flow (gas exchange) across stomata. The model is employed to explain the responses of plants to water deficit and other stresses. During water deficit, the cohesion-tension mechanism of the ascent of sap in the xylem plays an important role. Results are summarized which prove the validity of the coehesion/tension theory. Effects of the stress hormone abscisic acid (ABA) are presented. They show that there is an apoplastic component of the flow of ABA in the root which contributes to the ABA signal in the xylem. On the other hand, (+)-cis-trans-ABA specifically affects both the cell level (water channel activity) and water flow driven by gradients in osmotic pressure at the root level which is in agreement with the composite transport model. Hydraulic water flow in the presence of gradients in hydrostatic pressure remains unchanged. The results agree with the composite transport model and resemble earlier findings of high salinity obtained for the cell (Lp) and root (Lp(r)) level. They are in line with known effects of nutrient deprivation on root Lp(r )and the diurnal rhythm of root Lp(r )recently found in roots of LOTUS. PMID- 11006305 TI - Adaptation of roots to low water potentials by changes in cell wall extensibility and cell wall proteins. AB - It is common for the root/shoot ratio of plants to increase when water availability is limiting. This ratio increases because roots are less sensitive than shoots to growth inhibition by low water potentials. The physiological and molecular mechanisms that assist root growth under drought conditions are reviewed, with a focus on changes in cell walls. Maize seedlings adapt to low water potential by making the walls in the apical part of the root more extensible. In part, this is accomplished by increases in expansin activity and in part by other, more complex changes in the wall. The role of xyloglucan endotransglycosylase, peroxidase and other wall enzymes in root adaptation to low water potential is evaluated and some of the complications in the field of study are listed. PMID- 11006306 TI - Growth of Arabidopsis thaliana seedlings under water deficit studied by control of water potential in nutrient-agar media. AB - We have characterized the growth responses of Arabidopsis thaliana seedlings to water deficit. To manipulate the water potential, we developed a method whereby the nutrient-agar medium could be supplemented with polyethylene glycol (PEG 8000); PEG was introduced into gelled media by diffusion, which produced media with water potential as low as -1.6 MPa. For dark-grown plants, hypocotyl growth had a hyperbolic dependence on water potential, and was virtually stopped by -1 MPa. In contrast, primary root elongation was stimulated by moderate deficit and even at -1.6 MPa was not significantly less than the control. That these results did not depend on a direct effect of PEG was attested by obtaining indistinguishable results when a dialysis membrane impermeable to PEG was placed between the medium and the seedlings. For light-grown seedlings, moderate deficit also stimulated primary root elongation and severe deficit reduced elongation only partially. These changes in elongation were paralleled by changes in root system dry weight. At moderate deficit, lateral root elongation and initiation were unaffected and at higher stress levels both were inhibited. Primary root diameter increased steadily with time in well-watered controls and under water deficit increased transiently before stabilizing at a diameter that was inversely proportional to the deficit. Along with stimulated primary root elongation, moderate water deficit also stimulated the rate of cell production. Thus, A. thaliana responds to water deficit vigorously, which enhances its use as a model to uncover mechanisms underlying plant responses to water deficit. PMID- 11006307 TI - Control of abscisic acid synthesis. AB - The abscisic acid (ABA) biosynthetic pathway involves the formation of a 9-cis epoxycarotenoid precursor. Oxidative cleavage then results in the formation of xanthoxin, which is subsequently converted to ABA. A number of steps in the pathway may control ABA synthesis, but particular attention has been given to the enzyme involved in the oxidative cleavage reaction, i.e. 9-cis-epoxycarotenoid dioxygenase (NCED). Cloning of a gene encoding this enzyme in maize was first reported in 1997. Mapping and DNA sequencing studies indicated that a wilty tomato mutant was due to a deletion in the gene encoding an enzyme with a very similar amino acid sequence to this maize NCED. The potential use of this gene in altering ABA content will be discussed together with other genes encoding ABA biosynthetic enzymes. PMID- 11006308 TI - Endogenous ABA maintains shoot growth in tomato independently of effects on plant water balance: evidence for an interaction with ethylene. AB - To examine whether the reduced shoot growth of abscisic acid (ABA)-deficient mutants of tomato is independent of effects on plant water balance, flacca and notabilis were grown under controlled-humidity conditions so that their leaf water potentials were equal to or higher than those of well-watered wild-type plants throughout development. Most parameters of shoot growth remained markedly impaired and root growth was also greatly reduced. Additional experiments with flacca showed that shoot growth substantially recovered when wild-type levels of ABA were restored by treatment with exogenous ABA, even though improvement in leaf water potential was prevented. The ability of applied ABA to increase growth was greatest for leaf expansion, which was restored by 75%. The ethylene evolution rate of growing leaves was doubled in flacca compared to the wild type and treatment with silver thiosulphate to inhibit ethylene action partially restored shoot growth. The results demonstrate that normal levels of endogenous ABA are required to maintain shoot development, particularly leaf expansion, in well-watered tomato plants, independently of effects on plant water balance. The impairment of shoot growth caused by ABA deficiency is at least partly attributable to ethylene. PMID- 11006309 TI - The effects of ABA on channel-mediated K(+) transport across higher plant roots. AB - The transport and accumulation of K(+) in higher plant roots is regulated by ABA. Molecular and electrophysiological techniques have identified a number of discrete transporters which are involved in the translocation of K(+) from the soil solution to the shoots of higher plants. Furthermore, recent reports have shown that ABA regulates K(+) channel activity in maize and Arabidopsis roots which suggests that ABA regulation of K(+) transport in roots is, at least in part, ion channel-mediated. The signalling processes which underlie the ABA regulation of K(+) channels have been investigated. The effects of ABA on the membrane potential of intact maize root cells were also studied. It was found that ABA regulated the membrane potential of root cells and that this regulation is consistent with the hypothesis that ABA-induced K(+) accumulation in roots is mediated by K(+) channels. PMID- 11006310 TI - Sensitivity of growth of roots versus leaves to water stress: biophysical analysis and relation to water transport. AB - Water transport is an integral part of the process of growth by cell expansion and accounts for most of the increase in cell volume characterizing growth. Under water deficiency, growth is readily inhibited and growth of roots is favoured over that of leaves. The mechanisms underlying this differential response are examined in terms of Lockhart's equations and water transport. For roots, when water potential (psi) is suddenly reduced, osmotic adjustment occurs rapidly to allow partial turgor recovery and re-establishment of psi gradient for water uptake, and the loosening ability of the cell wall increases as indicated by a rapid decline in yield-threshold turgor. These adjustments permit roots to resume growth under low psi. In contrast, in leaves under reductions in psi of similar magnitude, osmotic adjustment occurs slowly and wall loosening ability either does not increase substantially or actually decreases, leading to marked growth inhibition. The growth region of both roots and leaves are hydraulically isolated from the vascular system. This isolation protects the root from low psi in the mature xylem and facilitates the continued growth into new moist soil volume. Simulations with a leaky cable model that includes a sink term for growth water uptake show that growth zone psi is barely affected by soil water removal through transpiration. On the other hand, hydraulic isolation dictates that psi of the leaf growth region would be low and subjected to further reduction by high evaporative demand. Thus, a combination of transport and changes in growth parameters is proposed as the mechanism co-ordinating the growth of the two organs under conditions of soil moisture depletion. The model simulation also showed that roots behave as reversibly leaky cable in water uptake. Some field data on root water extraction and vertical profiles of psi in shoots are viewed as manifestations of these basic phenomena. Also discussed is the trade-off between high xylem conductance and strong osmotic adjustment. PMID- 11006311 TI - Regulation of leaf and fruit growth in plants growing in drying soil: exploitation of the plants' chemical signalling system and hydraulic architecture to increase the efficiency of water use in agriculture. AB - In this paper the nature of root-to-shoot signals in plants growing in drying soil is considered in the context of their commercial exploitation in tomato (Lycopersicon esculentum L.) and other crops. Recent findings are presented on the effects of partial root drying (PRD) in the production of a glasshouse tomato crop. These findings show how an understanding of both root-to-shoot signalling mechanisms and fruit hydraulic architecture may explain observed increases in fruit quality, the differential effects of PRD on vegetative and reproductive production and the incidence of blossom end rot. Evidence is provided to support the hypothesis that the success of PRD may lie, at least in part, in the relative chemical and hydraulic isolation of the tomato fruit. PMID- 11006312 TI - Hormonal changes induced by partial rootzone drying of irrigated grapevine. AB - Partial rootzone drying (PRD) is a new irrigation technique which improves the water use efficiency (by up to 50%) of wine grape production without significant crop reduction. The technique was developed on the basis of knowledge of the mechanisms controlling transpiration and requires that approximately half of the root system is always maintained in a dry or drying state while the remainder of the root system is irrigated. The wetted and dried sides of the root system are alternated on a 10-14 d cycle. Abscisic acid (ABA) concentration in the drying roots increases 10-fold, but ABA concentration in leaves of grapevines under PRD only increased by 60% compared with a fully irrigated control. Stomatal conductance of vines under PRD irrigation was significantly reduced when compared with vines receiving water to the entire root system. Grapevines from which water was withheld from the entire root system, on the other hand, show a similar reduction in stomatal conductance, but leaf ABA increased 5-fold compared with the fully irrigated control. PRD results in increased xylem sap ABA concentration and increased xylem sap pH, both of which are likely to result in a reduction in stomatal conductance. In addition, there was a reduction in zeatin and zeatin riboside concentrations in roots, shoot tips and buds of 60, 50 and 70%, respectively, and this may contribute to the reduction in shoot growth and intensified apical dominance of vines under PRD irrigation. There is a nocturnal net flux of water from wetter roots to the roots in dry soil and this may assist in the distribution of chemical signals necessary to sustain the PRD effect. It was concluded that a major effect of PRD is the production of chemical signals in drying roots that are transported to the leaves where they bring about a reduction in stomatal conductance. PMID- 11006313 TI - Expression of the adrenomedullin gene in epithelial ovarian cancer. AB - Adrenomedullin (AM) gene expression was analysed in 60 cases of epithelial ovarian cancer, (29 serous, 14 mucinous, 13 endometrioid, three clear cell, and one undifferentiated) using reverse transcription-polymerase chain reaction (RT PCR); 10 of the cases were of low malignant potential; 25 were stage I; three were stage II; 27 were stage III; and five were stage IV. The level of AM gene expression was described in terms of the relative yield of the AM gene to the beta2-microglobulin gene. AM gene expression ranged from 0.04 to 1.57 (median 0.36). The association between histological grade and AM gene expression was significant (P: = 0.027), however, the association with other clinico pathological features, i.e. patients' age at diagnosis, stage of disease, residual tumour mass after initial surgery, and histological subtype were not significant. Survival data were available for all patients and univariate Cox regression analysis showed that the AM gene expression was significantly associated with a poor prognosis (P: = 0.019). Immunohistochemical studies showed that AM was localized in the outer cell membrane or the cytoplasm of the carcinoma cells and in the endothelial cells of the tumour stroma. The AM gene expression level may play a key role in the biology of epithelial ovarian cancer and may define a more aggressive tumour phenotype. PMID- 11006314 TI - No evidence for mutations of the leptin or leptin receptor genes in women with polycystic ovary syndrome. AB - Polycystic ovarian syndrome (PCOS) is often associated with obesity and insulin resistance, both of which are features that are linked to the leptin and leptin receptor (LEPR) genes. Analysis of the leptin gene by sequencing samples from 38 well-characterized patients with PCOS revealed no mutations of the coding exons. In single-stranded conformational polymorphism (SSCP) analysis and subsequent sequencing of the LEPR gene revealed previously identified amino acid variants in exons 2, 4 and 12 as well as the pentanucleotide insertion in the 3'-untranslated region (3'-UTR). The allele frequencies of these polymorphisms did not differ from those in the general population, as assessed in 122 female controls. Compared with non-carriers, serum insulin concentrations tended to be lower in the carriers of the variant LEPR exon 12 allele as well as in the carriers of the variant LEPR 3'-UTR allele, a marker previously suggested to be associated with serum insulin concentrations. In conclusion, PCOS is not commonly a consequence of mutations of the leptin or LEPR genes. However, our data support the hypothesis that variations in the LEPR gene locus have an effect on insulin regulation. PMID- 11006315 TI - The effect of FSH on male germ cell survival and differentiation in vitro is mimicked by pentoxifylline but not insulin. AB - High concentrations of FSH have been shown to boost in-vitro differentiation of germ cells from men with normal spermatogenesis and from some patients with in vivo maturation arrest. This study shows that the differentiation-promoting effect of FSH is connected to protection against germ cell apoptosis and that both effects can be mimicked by the intracellular cyclic AMP (cAMP)-elevating drug pentoxifylline. On the other hand, a high concentration of insulin, supposed to act at the insulin-like growth factor I receptor, did not exert any effect either on differentiation or apoptosis of germ cells in vitro. These data show that the in-vitro effects of supraphysiological concentrations of FSH on human spermatogenesis are mediated by the classical FSH signal transduction pathway involving cAMP as a second messenger. Pentoxifylline may thus be useful as an alternative means for intracellular cAMP elevation in men with high circulating FSH concentrations leading to desensitization of the FSH receptor. PMID- 11006316 TI - Prostasomes inhibit the NADPH oxidase activity of human neutrophils. AB - Prostasomes are particular lipid vesicles secreted by the prostate in human semen and involved in several physiological functions such as the improvement of sperm motility or immunomodulation. We have previously shown that they reduced the overall reactive oxygen species (ROS) production of seminal polymorphonuclear neutrophils (PMN). The present study was conducted to define the mechanism by which prostasomes inhibit the ROS production of blood and seminal PMN. The luminol chemiluminescence measuring total ROS production of blood PMN stimulated by either a phorbol ester (PMA) or a chemoattractant peptide, formyl-Met-Leu-Phe (fMLP) was significantly inhibited by prostasomes. The NADPH oxidase activity of the PMN was measured by 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1, 2 a]pyrazin-3-one (MCLA) chemiluminescence. Prostasomes inhibited the NADPH oxidase activity of blood or seminal PMN and increased the lag-phase of the enzyme after PMA stimulation. Prostasomes also inhibited significantly the NADPH oxidase activity of fMLP stimulated blood PMN, but the inhibition was not significant for seminal PMN. The lipid composition of blood PMN was analysed and compared to the lipid composition of prostasomes. This showed that prostasomes had a high cholesterol:phospholipid molar ratio and a high proportion of sphingomyelin. Together with the fact that prostasomes can rigidify the plasma membrane of blood PMN, these results led us to postulate that prostasomes inhibit the NADPH oxidase activity of PMN by lipid transfer from the prostasomes to the plasma membrane of the PMN. PMID- 11006317 TI - Acrosome reaction of human spermatozoa is mainly mediated by alpha1H T-type calcium channels. AB - The objectives of this study were: (i) to investigate the possible role of T-type Ca(2+) channels on the acrosome reaction (AR) of human spermatozoa; and (ii) to determine the sub-type of T-type calcium channels involved in the AR. The AR was induced in vitro by mannose-bovine serum albumin (BSA). The inhibitory effects of mibefradil (T-type Ca(2+) channel blocker), NiCl(2), or nifedipine (L-type Ca(2+) channel blocker) on the mannose-BSA induced AR were evaluated in capacitated human spermatozoa. The AR was sensitively inhibited by low micromolar concentrations of mibefradil (IC(50) = 1 micromol/l) in a dose-dependent manner. Low concentrations of Ni(2+) (IC(50) = 40 micromol/l) also inhibited the mannose BSA induced AR. On the contrary, higher concentrations of nifedipine were required to block AR (IC(50) = 60 micromol/l). Reverse transcription-polymerase chain reaction (RT-PCR) was performed to identify the sub-types of T-type channels present in human testes. Analysis of PCR products showed that only alpha1H subunits are expressed in testes. The expression of the alpha1H subunit may be tissue specific since its mRNA was not detected in the human ovary. The present study suggests that the AR of human spermatozoa is highly associated with T-type Ca(2+) channels and is mainly mediated by calcium influx through alpha1H T type Ca(2+) channels. PMID- 11006318 TI - In-vitro studies of the potential role of neutrophils in the process of menstruation. AB - Significant numbers of neutrophils are found extravascularly within the endometrium only during the immediate premenstrual and menstrual phases of the cycle. In this study we investigated the effect of neutrophil products on the synthesis and activation of matrix metalloproteinases (MMP), enzymes considered to play a crucial role in the degradation of endometrial tissue that occurs at menstruation. Latent MMP-2, MMP-3 and MMP-9 released by endometrial stromal fibroblasts and peripheral blood neutrophils were activated when the two cell types were cultured together. Tissue inhibitors of metalloproteinases (TIMP) 1 and 2 were also degraded in this system. Neutralization studies identified a role for the serine protease, elastase, in the observed activation of MMP. Although cultured endometrial neutrophils behaved similarly to peripheral blood neutrophils in their ability to release latent MMP-9 and elastase, no active forms of MMP-2. MMP-3 and MMP-9 were detected in supernatant from co-cultures containing endometrial neutrophils and stromal fibroblasts. This appeared to be due to an alteration in the neutrophil production of elastase and inhibitors. e.g. alpha1-antitrypsin, in these cultures so that active elastase was not available. Our results demonstrate that any involvement of neutrophils in the tissue destruction occurring at menstruation may be tightly regulated by the focal concentration of degradative enzymes and their respective inhibitors. PMID- 11006319 TI - Expression of interleukin-11 during the human menstrual cycle: coincidence with stromal cell decidualization and relationship to leukaemia inhibitory factor and prolactin. AB - Interleukin-11 (IL-11) is crucial in the decidualization response of the uterine stroma to the implanting blastocyst in the mouse. This study examined the localization and expression of IL-11 in human endometrium throughout the menstrual cycle and of prolactin and leukaemia inhibitory factor (LIF) in secretory phase endometrium. The mRNA expression of IL-11 receptor alpha and the signalling component, gp130, in endometrial tissue were also determined. Immunoreactive IL-11 was highest in the secretory phase and present in decidualized stromal cells, glandular epithelial cells, endothelial and smooth muscle cells, and the mRNA expression was verified by in-situ hybridization. Decidual cells showed the most intense staining. IL-11 receptor alpha and gp130 mRNA were detected throughout the cycle with minimal variation. Expression of IL 11 mRNA and protein preceded that of prolactin. While immunoreactive prolactin was found in stromal, decidual and glandular epithelial cells, prolactin mRNA was confined to decidual cells. In contrast, endometrial LIF expression preceded IL 11 but was largely confined to the glandular epithelium. The sequence of appearance of LIF, IL-11 and prolactin suggests a synchronized role for each in the differentiation of the endometrium. The cyclical changes and cell type specific expression of IL-11 suggests a potential role in the decidualization of stromal cells. PMID- 11006320 TI - Gene expression and tissue concentrations of IGF-I in human myometrium and fibroids under different hormonal conditions. AB - The expression of insulin-like growth factor-I (IGF-I) was measured at the mRNA and protein level in myometrium and fibroids from women with and without preoperative treatment with a gonadotrophin-releasing hormone (GnRH) agonist for 3 months, from post-menopausal women, from pregnant women and in myometrium from women without fibroid disease. Women with menstrual periods were classified according to the phase of the cycle. In tissues from non-treated premenopausal women, IGF-I mRNA expression was significantly higher in fibroids than in myometrium, with no differences related to phase of the menstrual cycle. In post menopausal women and in GnRH agonist-treated women responding to treatment, similar mRNA expression was seen in myometrium and fibroids but the concentrations were lower than in untreated premenopausal women. The IGF-I mRNA value in fibroids from pregnant women was higher than in any other group and myometrium from pregnant women exhibited higher mRNA expression than myometrium from non-treated premenopausal women. The IGF-I protein was more abundant in fibroids than in myometrium of non-treated premenopausal and of pregnant women and in both tissues the concentration was significantly higher in the group of pregnant women. The IGF-I protein concentrations in fibroids and myometrium from GnRH agonist-treated and post-menopausal women were similar to those from premenopausal non-treated women. High sex steroid concentrations in pregnant and non-pregnant women of fertile age seem to be associated with a higher expression of IGF-I in fibroids than in myometrium, suggesting that IGF-I contributes to the selective growth advantage of these tumours. PMID- 11006321 TI - Differential expression of the alpha(2)-macroglobulin receptor and the receptor associated protein in normal human endometrium and endometrial carcinoma. AB - Extracellular matrix degradation, mediated by the activation of receptor-bound proteolytic enzymes, is essential to the process of cellular invasion. Many normal physiological functions such as endometrial remodelling are reliant on the activation of these surface associated proteolytic enzymes, as are pathological functions such as cancer-cell invasion. The internalization of proteolytic complexes is mediated by the multi-functional clearance receptor, alpha(2) macroglobulin receptor/LRP. The role of LRP and its ligand binding inhibitor, the receptor-associated protein (RAP), in the advancement of invasive endometrial carcinoma is unknown. The aim of this study was to compare the expression of LRP and RAP mRNA in normal endometrium (n = 14) and endometrial carcinoma (n = 33) by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Expression of LRP mRNA in normal endometrium was significantly increased in the secretory phase when compared with proliferative phase endometrium (P: < 0.05). The expression of LRP in all carcinomas examined was significantly reduced to about 20% of the amount in normal endometrium (P: < 0.05), whereas RAP expression was not significantly different between endometrium and carcinoma. No significant difference in the level of LRP or RAP expression was observed between carcinoma grades or stages. In conclusion, we have shown that LRP expression is differentially regulated in the normal endometrium during the menstrual cycle and is decreased in invasive endometrial carcinomas. PMID- 11006322 TI - Increased concentrations of soluble tumour necrosis factor receptor (sTNFR) I and II in peritoneal fluid from women with endometriosis. AB - Tumour necrosis factor alpha (TNFalpha), a proapoptotic cytokine, is known to be present in peritoneal fluid from women with endometriosis. An emerging view is that soluble TNF receptors (sTNFR) can modulate the effects of TNFalpha by acting as TNFalpha antagonists. To assess the relevance of sTNFRs in the pathophysiology of endometriosis, concentrations of sTNFR I, sTNFR II and TNFalpha in peritoneal fluid from women with endometriosis (n = 53) and without endometriosis (n = 40) were measured. Concentrations of both sTNFR I and sTNFR II in peritoneal fluid from women with endometriosis were significantly higher than in peritoneal fluid from women without endometriosis, both in the follicular and the luteal phases. TNFalpha concentrations did not differ in patients with and without endometriosis in both phases. When stratified by the stage of the disease, women with both stages I/II and stages III/IV exhibited significantly higher concentrations of sTNFR I and sTNFR II in peritoneal fluid, compared with women without endometriosis, whereas no appreciable difference in the concentrations was detected between stages I/II and stages III/IV. A significant correlation was found between the concentrations of sTNFR I and sTNFR II; while the correlations between TNFalpha and sTNFR I or sTNFR II, were either not significant or were very weak. Furthermore, mRNA for the membrane-associated TNF receptor type 1 and TNF receptor type 2, both of which convey the effects of TNFalpha, were shown to be expressed in endometriotic tissues as well as eutopic endometrium. Together, these findings suggest a possible involvement of sTNFRs in the pathophysiology of endometriosis. PMID- 11006323 TI - Expression of vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) in conceptus and endometrium during implantation in the rhesus monkey. AB - The aim of this study was to analyse the expression of transcripts and proteins for vascular endothelial growth factor (VEGF) and placenta growth factor (PlGF) in different compartments of the early conceptus at primary implantation sites during lacunar (n = 6), early villous (n = 9) and villous placenta (n = 6) stages of gestation in the rhesus monkey. During the lacunar stage, VEGF expression was observed in the cytotrophoblast cells lining the extraembryonic cavity, but these cells did not express PlGF. With further development, cytotrophoblast cells lining villi, forming columns, and constituting anchoring villi, expressed both VEGF and PlGF during early villous and villous placenta stages. In addition, chorion, amnion and villous stromal cells expressed both VEGF and PlGF proteins and mRNA. During the lacunar stage, all epithelial cells in maternal endometrium generally expressed VEGF, while PlGF expression was observed in the plaque epithelium only. As gestation advanced, the expression of VEGF and PlGF from plaque cells decreased, and in surface and glandular epithelium the expression of VEGF increased, while the expression of PlGF remained unaltered. Decidual stromal cells expressed VEGF and PlGF only at low levels during the lacunar stage, while the expression of both increased during the early villous and the villous placenta stages of implantation. It appears from the present study that the expression of VEGF and PlGF are regulated in a temporal and spatial manner during early stages of implantation and that their concerted actions in placental and maternal compartments play a critical role in the evolving pregnancy in the rhesus monkey. PMID- 11006324 TI - Localization of E-cadherin in villous, extravillous and vascular trophoblasts during intrauterine, ectopic and molar pregnancy. AB - Previous reports have described down-regulation of E-cadherin in trophoblasts differentiating to an invasive phenotype. This study shows the localization of E cadherin in a prospective design with stereological sampling of fetal and maternal first, second and third trimester tissue. E-cadherin was observed in villous cytotrophoblasts, and in non-proliferating, intermediate trophoblasts (IT) within cell columns and islands in intrauterine, ectopic and partial molar placentas. Highly proliferating IT with cytological atypia in complete molar placentas were also E-cadherin-positive. E-cadherin was present in trophoblasts throughout the anchoring cell columns. Trophoblasts undergoing epithelial mesenchymal transformation (EMT) detaching from the distal cell columns and deeper located single extravillous interstitial trophoblasts (EVT) showed E cadherin-negative breaches in the cell membrane. Prior to the late second trimester, the relative number of E-cadherin-positive single EMT and EVT differed from the total number of cytokeratin-positive trophoblasts. Intraluminal, endovascular and perivascular trophoblasts adjacent to the maternal vessels were also E-cadherin-positive, but a highly varying pattern was observed at different ages of gestation. Our results indicate a temporary shift in E-cadherin expression in extravillous trophoblasts possessing a migrating and invasive potential. Functional E-cadherin may be restored as trophoblasts aggregate in the decidua and the vessel wall after completion of migration. PMID- 11006325 TI - Leptin modulates extracellular matrix molecules and metalloproteinases: possible implications for trophoblast invasion. AB - Leptin is a circulating hormone which plays an important role in the regulation of energy balance, haemopoiesis and reproduction. Leptin and its receptor (leptin R) are localized in human placental tissue but their function is not known. In this study we have investigated the expression of leptin and leptin-R in the human placenta with particular attention to extravillous cytotrophoblastic cell islands and cell columns which play a pivotal role in trophoblast invasion and placental growth. We demonstrate that leptin-R immunoreactivity shows a strong expression in the distal extravillous cytotrophoblastic cells of cell columns invading the basal plate, whereas leptin expression is homogeneously expressed in all the cellular components of cell columns. Since the invasive ability of the distally located extravillous cytotrophoblast of cell columns is known to be regulated by a variety of proteases and some extracellular matrix molecules, we tested the influence of leptin on the in-vitro production of matrix metalloproteinase (MMP)-2, MMP-9 and fetal fibronectin (fFN) by cytotrophoblastic cells. We demonstrate that leptin increases, in a dose-dependent manner, the secretion of immunoreactive MMP-2 and fFN and enhances the activity of MMP-9 in cultured cytotrophoblastic cells. Our results suggest that leptin and leptin-R could have a role in the invasive processes of the extravillous cytotrophoblastic cells by modulating the expression of MMPs. In addition, these results provide a foundation for studying pathological conditions characterized by insufficient or excessive trophoblast invasion. PMID- 11006326 TI - Local fetal signal is not required for maintaining IGFBP gene expression in the human decidua: evidence from extrauterine pregnancies. AB - Insulin-like growth factor-II (IGF-II) from the invading extravillous cytotrophoblasts (EVTs) and insulin-like growth factor binding proteins (IGFBPs) from the maternal decidua interact at the feto-maternal interface and regulate implantation and placentation. To determine whether a local stimulus from the fetus is important in the regulation of IGFBP gene expression in the human decidua, we compared the expression of IGFBP genes in intra- and extrauterine (tubal) pregnancies. The expression of IGF-II and IGFBP-1 to IGFBP-6 mRNAs was determined by in-situ hybridization in the Fallopian tubes of extrauterine pregnancies and concurrent decidua (n = 6), and in the placentae and Fallopian tubes of intrauterine pregnancies (n = 6). All six IGFBP mRNAs were identified in the decidualized endometrium and decidualized Fallopian tubes of intra- and extrauterine pregnancies, with IGFBP-1 mRNA being the predominant mRNA. IGFBP-4 was the second most predominant mRNA and was slightly more abundant in the decidua of extrauterine pregnancies than of intrauterine pregnancies. IGF-II mRNA was expressed mainly in cells of fetal origin. The fact that the IGFBP mRNAs were expressed similarly in both intra- and extrauterine pregnancies indicates that the local physical stimulus from an implanting fetus is not necessary to induce or maintain decidual IGFBP gene expression. PMID- 11006327 TI - Plant PtdIns 3-kinase goes nuclear. PMID- 11006328 TI - Cell biology of plant and fungal tip growth--getting to the point. PMID- 11006329 TI - Comparative genomics of plant chromosomes. PMID- 11006330 TI - Overexpression of a gene encoding a cytochrome P450, CYP78A9, induces large and seedless fruit in arabidopsis. AB - An activation tagging screen in which the cauliflower mosaic virus 35S enhancer was inserted randomly into an Arabidopsis genome homozygous for the floral homeotic mutation apetala2-1 (ap2-1) resulted in a line (28-5) with extraordinarily wide, heart-shaped ovaries. The ovary of the 28-5 ap2-1 mutant shows an oval shape because of increased numbers of enlarged cells. When the ap2 1 mutation is crossed out of the genetic background, more elongated rather than wider fruits are obtained. Normally, Arabidopsis fruits will develop to a normal size only when the ovules are present and fertilized. In the 28-5 single mutant, the siliques keep growing despite failure of fertilization and can reach nearly normal size. When wild-type pollen was used to pollinate the mutant pistil, the pollinated 28-5 silique became >10% longer and 40% wider than a wild-type silique, although producing very few seeds. The enhancer insertion in line 28-5 acts by hyperactivating a cytochrome P450 gene, CYP78A9. The pistil of 28-5 ap2-1 mutant flowers shows a structure similar to that of Capsella bursa-pastoris, a distant mustard relative of Arabidopsis, suggesting that the processes regulated by the CYP78A9-encoded protein may be involved in evolutionary control of carpel shape. PMID- 11006331 TI - Phenotypic instability and rapid gene silencing in newly formed arabidopsis allotetraploids. AB - Allopolyploid hybridization serves as a major pathway for plant evolution, but in its early stages it is associated with phenotypic and genomic instabilities that are poorly understood. We have investigated allopolyploidization between Arabidopsis thaliana (2n = 2x = 10; n, gametic chromosome number; x, haploid chromosome number) and Cardaminopsis arenosa (2n = 4x = 32). The variable phenotype of the allotetraploids could not be explained by cytological abnormalities. However, we found suppression of 20 of the 700 genes examined by amplified fragment length polymorphism of cDNA. Independent reverse transcription polymerase chain reaction analyses of 10 of these 20 genes confirmed silencing in three of them, suggesting that approximately 0.4% of the genes in the allotetraploids are silenced. These three silenced genes were characterized. One, called K7, is repeated and similar to transposons. Another is RAP2.1, a member of the large APETALA2 (AP2) gene family, and has a repeated element upstream of its 5' end. The last, L6, is an unknown gene close to ALCOHOL DEHYDROGENASE on chromosome 1. CNG DNA methylation of K7 was less in the allotetraploids than in the parents, and the element varied in copy number. That K7 could be reactivated suggests epigenetic regulation. L6 was methylated in the C. arenosa genome. The present evidence that gene silencing accompanies allopolyploidization opens new avenues to this area of research. PMID- 11006332 TI - UV radiation-sensitive norin 1 rice contains defective cyclobutane pyrimidine dimer photolyase. AB - Norin 1, a progenitor of many economically important Japanese rice strains, is highly sensitive to the damaging effects of UVB radiation (wavelengths 290 to 320 nm). Norin 1 seedlings are deficient in photorepair of cyclobutane pyrimidine dimers. However, the molecular origin of this deficiency was not known and, because rice photolyase genes have not been cloned and sequenced, could not be determined by examining photolyase structural genes or upstream regulatory elements for mutations. We therefore used a photoflash approach, which showed that the deficiency in photorepair in vivo resulted from a functionally altered photolyase. These results were confirmed by studies with extracts, which showed that the Norin 1 photolyase-dimer complex was highly thermolabile relative to the wild-type Sasanishiki photolyase. This deficiency results from a structure/function alteration of photolyase rather than of nonspecific repair, photolytic, or regulatory elements. Thus, the molecular origin of this plant DNA repair deficiency, resulting from a spontaneously occurring mutation to UV radiation sensitivity, is defective photolyase. PMID- 11006333 TI - A GAL4-like protein is involved in the switch between biotrophic and necrotrophic phases of the infection process of Colletotrichum lindemuthianum on common bean. . AB - Random insertional mutagenesis was conducted with the hemibiotrophic fungus Colletotrichum lindemuthianum, causal agent of common bean anthracnose. Nine mutants that were altered in their infection process on the host plant were generated. One of these, H433 is a nonpathogenic mutant able to induce necrotic spots on infected leaves rapidly. These spots are similar to those observed during the hypersensitive reaction. Cytological observations showed that the development of the mutant H433 is stopped at the switch between the biotrophic and the necrotrophic phases. This mutant carries two independent insertions of the transforming plasmid pAN7-1. Complementation studies using the wild-type genomic regions corresponding to the two insertions showed that one is responsible for the H433 phenotype. Sequencing analysis identified a single open reading frame that encoded a putative transcriptional activator belonging to the fungal zinc cluster (Zn[II](2)Cys(6)) family. The corresponding gene was designated CLTA1 (for C. lindemuthianum transcriptional activator 1). Expression studies showed that CLTA1 is expressed in low amounts during in vitro culture. Targeted disrupted strains were generated, and they exhibited the same phenotype as the original mutant H433. Complementation of these disrupted strains by the CLTA1 gene led to full restoration of pathogenicity. This study demonstrates that CLTA1 is both a pathogenicity gene and a regulatory gene involved in the switch between biotrophy and necrotrophy of the infection process of a hemibiotrophic fungus. PMID- 11006334 TI - Loss of function of a rice brassinosteroid insensitive1 homolog prevents internode elongation and bending of the lamina joint. AB - Brassinosteroids (BRs) are plant growth-promoting natural products required for plant growth and development. Physiological studies have demonstrated that exogenous BR, alone or in combination with auxin, enhance bending of the lamina joint of rice. However, little is known about the function of endogenous BR in rice or other grass species. We report here the phenotypical and molecular characterization of a rice dwarf mutant, d61, that is less sensitive to BR compared to the wild type. We cloned a rice gene, OsBRI1, with extensive sequence similarity to that of the Arabidopsis BRI gene, which encodes a putative BR receptor kinase. Linkage analysis showed that the OsBRI1 gene is closely linked to the d61 locus. Single nucleotide substitutions found at different sites of the d61 alleles would give rise to amino acid changes in the corresponding polypeptides. Furthermore, introduction of the entire OsBRI1 coding region, including the 5' and 3' flanking sequences, into d61 plants complemented the mutation to display the wild-type phenotype. Transgenic plants carrying the antisense strand of the OsBRI1 transcript showed similar or even more severe phenotypes than those of the d61 mutants. Our results show that OsBRI1 functions in various growth and developmental processes in rice, including (1) internode elongation, by inducing the formation of the intercalary meristem and the longitudinal elongation of internode cells; (2) bending of the lamina joint; and (3) skotomorphogenesis. PMID- 11006335 TI - Regulation of SUP expression identifies multiple regulators involved in arabidopsis floral meristem development. AB - During the course of flower development, floral homeotic genes are expressed in defined concentric regions of floral meristems called whorls. The SUPERMAN (SUP, also called FLO10) gene, which encodes a C2H2-type zinc finger protein, is involved in maintenance of the stamen/carpel whorl boundary (the boundary between whorl 3 and whorl 4) in Arabidopsis. Here, we show that the regulation of SUP expression in floral meristems is complex, consisting of two distinct phases, initiation and maintenance. The floral meristem identity gene LEAFY (LFY) plays a role in the initiation phase through at least two pathways, which differ from each other in the involvement of two homeotic genes, APETALA3 (AP3) and PISTILLATA (PI). AP3, PI, and another homeotic gene, AGAMOUS (AG), are further required for SUP expression in the later maintenance phase. Aside from these genes, there are other as yet unidentified genes that control both the temporal and spatial patterns of SUP expression in whorl 3 floral meristems. SUP appears to act transiently, probably functioning to trigger a genetic circuit that creates the correct position of the whorl 3/whorl 4 boundary. PMID- 11006336 TI - anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. AB - The petunia loci anthocyanin1 (an1), an2, an4, and an11 are required for the transcription of anthocyanin biosynthetic genes in floral organs. The an2 and an11 loci were recently cloned and shown to encode a MYB-domain transcriptional activator and a cytosolic WD40 protein, respectively. Here, we report the isolation of an1 by transposon tagging. an1 encodes a new member of the basic helix-loop-helix family of transcription factors that is functionally and evolutionarily distinct from JAF13, the apparent petunia ortholog of maize RED1 and snapdragon DELILA. We provide genetic evidence that the transcription factors encoded by an1, an2, and an4 operate in an unexpectedly complex regulatory hierarchy. In leaves, ectopic expression of AN2 induces an1 expression, whereas in anthers, an1 expression depends on an4, encoding (or controlling) a MYB protein that is paralogous to AN2. Experiments with transgenic plants expressing a post-translationally controlled AN1-GLUCOCORTICOID RECEPTOR fusion protein indicated that independent of protein synthesis, AN1 directly activates the expression of the dfrA gene encoding the enzyme dihydroflavonol 4-reductase and of Pmyb27 encoding a MYB-domain protein of unknown function. PMID- 11006337 TI - Jasmonic acid signaling modulates ozone-induced hypersensitive cell death. AB - Recent studies suggest that cross-talk between salicylic acid (SA)-, jasmonic acid (JA)-, and ethylene-dependent signaling pathways regulates plant responses to both abiotic and biotic stress factors. Earlier studies demonstrated that ozone (O(3)) exposure activates a hypersensitive response (HR)-like cell death pathway in the Arabidopsis ecotype Cvi-0. We now have confirmed the role of SA and JA signaling in influencing O(3)-induced cell death. Expression of salicylate hydroxylase (NahG) in Cvi-0 reduced O(3)-induced cell death. Methyl jasmonate (Me JA) pretreatment of Cvi-0 decreased O(3)-induced H(2)O(2) content and SA concentrations and completely abolished O(3)-induced cell death. Cvi-0 synthesized as much JA as did Col-0 in response to O(3) exposure but exhibited much less sensitivity to exogenous Me-JA. Analyses of the responses to O(3) of the JA-signaling mutants jar1 and fad3/7/8 also demonstrated an antagonistic relationship between JA- and SA-signaling pathways in controlling the magnitude of O(3)-induced HR-like cell death. PMID- 11006338 TI - Four genes of Medicago truncatula controlling components of a nod factor transduction pathway. AB - Rhizobium nodulation (Nod) factors are lipo-chitooligosaccharides that act as symbiotic signals, eliciting several key developmental responses in the roots of legume hosts. Using nodulation-defective mutants of Medicago truncatula, we have started to dissect the genetic control of Nod factor transduction. Mutants in four genes (DMI1, DMI2, DMI3, and NSP) were pleiotropically affected in Nod factor responses, indicating that these genes are required for a Nod factor activated signal transduction pathway that leads to symbiotic responses such as root hair deformations, expressions of nodulin genes, and cortical cell divisions. Mutant analysis also provides evidence that Nod factors have a dual effect on the growth of root hair: inhibition of endogenous (plant) tip growth, and elicitation of a novel tip growth dependent on (bacterial) Nod factors. dmi1, dmi2, and dmi3 mutants are also unable to establish a symbiotic association with endomycorrhizal fungi, indicating that there are at least three common steps to nodulation and endomycorrhization in M. truncatula and providing further evidence for a common signaling pathway between nodulation and mycorrhization. PMID- 11006339 TI - SOS3 function in plant salt tolerance requires N-myristoylation and calcium binding. AB - The salt tolerance gene SOS3 (for salt overly sensitive3) of Arabidopsis is predicted to encode a calcium binding protein with an N-myristoylation signature sequence. Here, we examine the myristoylation and calcium binding properties of SOS3 and their functional significance in plant tolerance to salt. Treatment of young Arabidopsis seedlings with the myristoylation inhibitor 2-hydroxymyristic acid caused the swelling of root tips, mimicking the phenotype of the salt hypersensitive mutant sos3-1. In vitro translation assays with reticulocyte showed that the SOS3 protein was myristoylated. Targeted mutagenesis of the N terminal glycine-2 to alanine prevented the myristoylation of SOS3. The functional significance of SOS3 myristoylation was examined by expressing the wild-type myristoylated SOS3 and the mutated nonmyristoylated SOS3 in the sos3-1 mutant. Expression of the myristoylated but not the nonmyristoylated SOS3 complemented the salt-hypersensitive phenotype of sos3-1 plants. No significant difference in membrane association was observed between the myristoylated and nonmyristoylated SOS3. Gel mobility shift and (45)Ca(2)+ overlay assays demonstrated that SOS3 is a unique calcium binding protein and that the sos3-1 mutation substantially reduced the capacity of SOS3 to bind calcium. The resulting mutant SOS3 protein was not able to interact with the SOS2 protein kinase and was less capable of activating it. Together, these results strongly suggest that both N-myristoylation and calcium binding are required for SOS3 function in plant salt tolerance. PMID- 11006340 TI - Association of phosphatidylinositol 3-kinase with nuclear transcription sites in higher plants. AB - The kinases responsible for phosphorylation of inositol-containing lipids are essential for many aspects of normal eukaryotic cell function. Genetic and biochemical studies have established that the phosphatidylinositol (PtdIns) 3 kinase encoded by the yeast VPS34 gene is essential for the efficient sorting and delivery of proteins to the vacuole; the kinase encoded by the human VPS34 homolog has been equally implicated in the control of intracellular vesicle traffic. The plant VPS34 homolog also is required for normal growth and development, and although a role for PtdIns 3-kinase in vesicle trafficking is likely, it has not been established. In this study, we have shown that considerable PtdIns 3-kinase activity is associated with the internal matrix of nuclei isolated from carrot suspension cells. Immunocytochemical and confocal laser scanning microscopy studies using the monoclonal antibody JIM135 (John Innes Monoclonal 135), raised against a truncated version of the soybean PtdIns 3 kinase, SPI3K-5p, revealed that this kinase appears to have a distinct and punctate distribution within the plant nucleus and nucleolus. Dual probing of root sections with JIM135 and anti-bromo-UTP antibodies, after in vitro transcription had been allowed to proceed in the presence of bromo-UTP, showed that SPI3K-5p associates with active nuclear and nucleolar transcription sites. These findings suggest a possible link between PtdIns 3-kinase activity and nuclear transcription in plants. PMID- 11006341 TI - Genetic manipulation of isoflavone 7-O-methyltransferase enhances biosynthesis of 4'-O-methylated isoflavonoid phytoalexins and disease resistance in alfalfa. AB - 4'-O-Methylation of an isoflavonoid intermediate is a key reaction in the biosynthesis of the phytoalexin medicarpin in legumes. However, isoflavone O methyltransferase (IOMT) from alfalfa converts the isoflavone daidzein to 7-O methyl daidzein (isoformononetin) in vitro as well as in vivo in unchallenged leaves of transgenic alfalfa ectopically expressing IOMT. In contrast, elicitation of IOMT-overexpressing plants with CuCl(2) or infecting these plants with Phoma medicaginis leads to greater accumulation of formononetin (4'-O-methyl daidzein) and medicarpin in the leaves than does elicitation or infection of control plants, and no isoformononetin is detected. Overexpression of IOMT results in increased induction of phenylpropanoid/isoflavonoid pathway gene transcripts after infection but has little effect on basal expression of these genes. IOMT-overexpressing plants display resistance to P. medicaginis. The apparently different regiospecificities of IOMT in vivo and in vitro are discussed in relation to potential metabolic channeling at the entry point into the isoflavonoid pathway. PMID- 11006342 TI - MAP kinase and protein kinase A-dependent mobilization of triacylglycerol and glycogen during appressorium turgor generation by Magnaporthe grisea. AB - Magnaporthe grisea produces an infection structure called an appressorium, which is used to breach the plant cuticle by mechanical force. Appressoria generate hydrostatic turgor by accumulating molar concentrations of glycerol. To investigate the genetic control and biochemical mechanism for turgor generation, we assayed glycerol biosynthetic enzymes during appressorium development, and the movement of storage reserves was monitored in developmental mutants. Enzymatic activities for glycerol generation from carbohydrate sources were present in appressoria but did not increase during development. In contrast, triacylglycerol lipase activity increased during appressorium maturation. Rapid glycogen degradation occurred during conidial germination, followed by accumulation in incipient appressoria and dissolution before turgor generation. Lipid droplets also moved to the incipient appressorium and coalesced into a central vacuole before degrading at the onset of turgor generation. Glycogen and lipid mobilization did not occur in a Deltapmk1 mutant, which lacked the mitogen activated protein kinase (MAPK) required for appressorium differentiation, and was retarded markedly in a DeltacpkA mutant, which lacks the catalytic subunit of cAMP-dependent protein kinase A (PKA). Glycogen and lipid degradation were very rapid in a Deltamac1 sum1-99 mutant, which carries a mutation in the regulatory subunit of PKA, occurring before appressorium morphogenesis was complete. Mass transfer of storage carbohydrate and lipid reserves to the appressorium therefore occurs under control of the PMK1 MAPK pathway. Turgor generation then proceeds by compartmentalization and rapid degradation of lipid and glycogen reserves under control of the CPKA/SUM1-encoded PKA holoenzyme. PMID- 11006343 TI - Identification and characterization of a novel microtubule-based motor associated with membranous organelles in tobacco pollen tubes. AB - Pollen tube growth depends on the differential distribution of organelles and vesicles along the tube. The role of microtubules in organelle movement is uncertain, mainly because information at the molecular level is limited. In an effort to understand the molecular basis of microtubule-based movement, we isolated from tobacco pollen tubes polypeptides that cosediment with microtubules in an ATP-dependent manner. Major polypeptides released from microtubules by ATP (ATP-MAPs) had molecular masses of 90, 80, and 41 kD. Several findings indicate that the 90-kD ATP-MAP is a kinesin-related motor: binding of the polypeptide to microtubules was enhanced by the nonhydrolyzable ATP analog AMP-PNP; the 90-kD polypeptide reacted specifically with a peptide antibody directed against a highly conserved region in the motor domain of the kinesin superfamily; purified 90-kD ATP-MAP induced microtubules to glide in motility assays in vitro; and the 90-kD ATP-MAP cofractionated with microtubule-activated ATPase activity. Immunolocalization studies indicated that the 90-kD ATP-MAP binds to organelles associated with microtubules in the cortical region of the pollen tube. These findings suggest that the 90-kD ATP-MAP is a kinesin-related microtubule motor that moves organelles in the cortex of growing pollen tubes. PMID- 11006344 TI - A lily stylar pectin is necessary for pollen tube adhesion to an in vitro stylar matrix. AB - Pollen tube cells adhere to the wall surface of the stylar transmitting tract epidermis in lily. This adhesion has been proposed as essential for the proper delivery of the sperm cells to the ovule. An in vitro adhesion bioassay has been used to isolate two stylar molecules required for lily pollen tube adhesion. The first molecule was determined to be a small, cysteine-rich protein with some sequence similarity to lipid transfer proteins and now called stigma/stylar cysteine-rich adhesin (SCA). The second, larger, molecule has now been purified from style fragments and characterized. Chemical composition, specific enzyme degradations, and immunolabeling data support the idea that this molecule required for pollen tube adhesion is a pectic polysaccharide. In vitro binding assays revealed that this lily stylar adhesive pectin and SCA are able to bind to each other in a pH-dependent manner. PMID- 11006345 TI - The classical arabinogalactan protein gene family of arabidopsis. AB - Arabinogalactan proteins (AGPs) are extracellular proteoglycans implicated in plant growth and development. We searched for classical AGPs in Arabidopsis by identifying expressed sequence tags based on the conserved domain structure of the predicted protein backbone. To confirm that these genes encoded bona fide AGPs, we purified native AGPs and then deglycosylated and deblocked them for N terminal protein sequencing. In total, we identified 15 genes encoding the protein backbones of classical AGPs, including genes for AG peptides-AGPs with very short backbones (10 to 13 amino acid residues). Seven of the AGPs were verified as AGPs by protein sequencing. A gene encoding a putative cell adhesion molecule with AGP-like domains was also identified. This work provides a firm foundation for beginning functional analysis by using a genetic approach. PMID- 11006346 TI - Biogenesis of the chloroplast-encoded D1 protein: regulation of translation elongation, insertion, and assembly into photosystem II. AB - Regulation of translation elongation, membrane insertion, and assembly of the chloroplast-encoded D1 protein of photosystem II (PSII) was studied using a chloroplast translation system in organello. Translation elongation of D1 protein was found to be regulated by (1) a redox component that can be activated not only by photosynthetic electron transfer but also by reduction with DTT; (2) the trans thylakoid proton gradient, which is absolutely required for elongation of D1 nascent chains on the thylakoid membrane; and (3) the thiol reactants N ethylmaleimide (NEM) and iodosobenzoic acid (IBZ), which inhibit translation elongation with concomitant accumulation of distinct D1 pausing intermediates. These results demonstrate that D1 translation elongation and membrane insertion are tightly coupled and highly regulated processes in that proper insertion is a prerequisite for translation elongation of D1. Cotranslational and post translational assembly steps of D1 into PSII reaction center and core complexes occurred independently of photosynthetic electron transfer or trans-thylakoid proton gradient but were strongly affected by the thiol reactants DTT, NEM, and IBZ. These compounds reduced the stability of the early PSII assembly intermediates, hampered the C-terminal processing of the precursor of D1, and prevented the post-translational reassociation of CP43, indicating a strong dependence of the D1 assembly steps on proper redox conditions and the formation of disulfide bonds. PMID- 11006348 TI - PPAR-alpha: a key to the mechanism of hepatoprotection by clofibrate. AB - The article highlighted in this issue is "Peroxisome Proliferator-Activated Receptor Alpha-Null Mice Lack Resistance to Acetaminophen Hepatotoxicity Following Clofibrate Exposure" by Chuan Chen, Gayle E. Hennig, Herbert E. Whiteley, J Christopher Corton, and Jose E. Manautou (pp. 338-344). PMID- 11006347 TI - The bacterial elicitor flagellin activates its receptor in tomato cells according to the address-message concept. AB - flg22, a peptide corresponding to the most conserved domain of bacterial flagellin, acts as a potent elicitor in plants. Here, we have used an iodinated derivative of flg22 ((125)I-labeled Tyr-flg22) as a molecular probe for the flagellin receptor in tomato cells. This radioligand showed rapid binding to a single class of specific, saturable, high-affinity receptor sites in intact cells and membrane preparations. Binding, although essentially nonreversible under physiological conditions, was not covalent, and chemical cross-linking was required to specifically label a single polypeptide of 115 kD. Intact flagellin and elicitor-active flagellin peptides but not biologically inactive analogs efficiently competed for binding of radioligand. Peptides lacking the C terminus of the conserved domain, previously found to act as competitive antagonists of elicitor action in tomato cells, also competed for binding of radioligand. Thus, this novel, high-affinity binding site exhibited all the characteristics expected of a functional receptor of bacterial flagellin. For a model of receptor activation, we propose a two-step mechanism according to the address-message concept, in which binding of the N terminus (address) is the first step and activation of responses with the C terminus (message) is the second step. PMID- 11006349 TI - Sheldon D. Murphy. PMID- 11006350 TI - Symposium overview: mechanism of action of nicotine on neuronal acetylcholine receptors, from molecule to behavior. AB - Nicotine has long been known to interact with nicotinic acetylcholine (ACh) receptors since Langley used it extensively to chart sympathetic ganglia a century ago. It has also been used as an effective insecticide. However, it was not until the 1990s that the significance of nicotine was increasingly recognized from the toxicological, pharmacological, and environmental points of view. This is partly because studies of neuronal nicotinic ACh receptors are rapidly emerging from orphan status, fueled by several lines of research. Since Alzheimer's disease is known to be associated with down-regulation of cholinergic activity in the brain, a variety of nicotine derivatives are being tested and developed for treatment of the disease. Public awareness of the adverse effects of nicotine has reached the highest level recently. Since insect resistance to insecticides is one of the most serious issues in the pest-control arena, it is an urgent requirement to develop new insecticides that act on target sites not shared by the existing insecticides. The neuronal nicotinic ACh receptor is one of them, and new nicotinoids are being developed. Thus, the time is ripe to discuss the mechanism of action of nicotine from a variety of angles, including the molecular, physiological, and behavioral points of view. This Symposium covered a wide area of nicotine studies: genetic, genomic, and functional aspects of nicotinic ACh receptors were studied, as related to anthelmintics and insecticides; interactions between ethanol and nicotine out the ACh receptor were analyzed, in an attempt to explain the well-known heavy drinker-heavy smoker correlation; the mechanisms that underlie the desensitization of ACh receptors were studied as related to nicotine action; selective pharmacological profiles of nicotine, and descriptions of some derivatives were described; and chronic nicotine infusion effects on memory were examined using animal models. PMID- 11006351 TI - Quantification of DNA adducts formed in liver, lungs, and isolated lung cells of rats and mice exposed to (14)C-styrene by nose-only inhalation. AB - Bronchiolo-alveolar tumors were observed in mice exposed chronically to 160 ppm styrene, whereas no tumors were seen in rats up to concentrations of 1000 ppm. Clara cells, which are predominant in the bronchiolo-alveolar region in mouse lungs but less numerous in rat and human lung, contain various cytochrome P450s, which may oxidize styrene to the rodent carcinogen styrene-7,8-oxide (SO) and other reactive metabolites. Reactive metabolites may form specific DNA adducts and induce the tumors observed in mice. To determine DNA adducts in specific tissues and cell types, rats and mice were exposed to 160 ppm [ring-U (14)C]styrene by nose-only inhalation for 6 h in a recirculating exposure system. Liver and lungs were isolated 0 and 42 h after exposure. Fractions enriched in Type II cells and Clara cells were isolated from rat and mouse lung, respectively. DNA adduct profiles differed quantitatively and qualitatively in liver, total lung, and enriched lung cell fractions. At 0 and 42 h after exposure, the two isomeric N:7-guanine adducts of SO (measured together, HPEG) were present in liver at 3.0 +/- 0.2 and 1.9 +/- 0.3 (rat) and 1.2 +/- 0.2 and 3.2 +/- 0.5 (mouse) per 10(8) bases. Several other, unidentified adducts were present at two to three times higher concentrations in mouse, but not in rat liver. In both rat and mouse lung, HPEG was the major adduct at approximately 1 per 10(8) bases at 0 h, and these levels halved at 42 h. In both rat Type II and non-Type II cells, HPEG was the major adduct and was about three times higher in Type II cells than in total lung. For mice, DNA adduct levels in Clara cells and non-Clara cells were similar to total lung. The hepatic covalent binding index (CBI) at 0 and 42 h was 0.19 +/- 0.06 and 0.14 +/- 0.03 (rat) and 0. 25 +/- 0.11 and 0.44 +/- 0.23 (mouse), respectively. The pulmonary CBIs, based on tissues combined for 0 and 42 h, were 0.17 +/- 0.04 (rat) and 0.24 +/- 0.04 (mouse). Compared with CBIs for other genotoxicants, these values indicate that styrene has only very weak adduct-forming potency. The overall results of this study indicate that DNA adduct formation does not play an important role in styrene tumorigenicity in chronically exposed mice. PMID- 11006352 TI - Influence of beta-naphthoflavone on 7,12-dimethylbenz(a)anthracene metabolism, DNA adduction, and tumorigenicity in rainbow trout. AB - Metabolism, DNA adduction, and tumor induction by 7, 12-dimethylbenz(a)anthracene (DMBA) were examined in cultured trout liver cells and in vivo in trout. Modulating CYP1A1 activity indicated this enzyme plays a significant role in metabolizing DMBA to water-soluble compounds in isolated trout liver cells. The major DMBA metabolites identified in trout liver cells were 10-, 11-, 8,9-, and 5,6-DMBA dihydrodiols, and DMBA, 2- or 3- or 4-phenol; 7-OH-methyl-12-methyl benz(a)anthracene and 12-OH-methyl-7-methyl-benz(a)anthracene were minor metabolites. A very small amount of DMBA-3,4-dihydrodiol was detected, and polar metabolites, which did not migrate with any DMBA metabolite standards, were observed. Incubating trout hepatocytes with DMBA-3, 4-dihydrodiol produced three prominent, nonpolar adducts indistinguishable from those in mouse embryo cells. However, DMBA-DNA adducts, formed in trout in vivo or in trout liver cells exposed to DMBA, were predominantly more polar than those formed in mouse embryo fibroblasts, and levels of DMBA-DNA adducts formed in trout liver cells were not significantly altered by modulating CYP1A1 activity. No significant repair of DMBA-DNA adducts was detected in cultured trout liver cells over a 48-h period, supporting previous studies indicating that fish are less efficient than mammals in repairing polyaromatic hydrocarbon DNA adducts. Compared to animals receiving DMBA alone, beta-naphthoflavone pretreatment in vivo did not affect hepatic CYP1A1, DMBA-DNA adducts, nor hepatic tumor response; but did significantly reduce tumor response in two other target organs. These results collectively indicate that DMBA bioactivation to DNA-binding metabolites in trout liver cells and mouse embryo cells predominantly involve different metabolic pathways to form the DNA-binding intermediates. PMID- 11006353 TI - Developmental and tissue-specific expression of AHR1, AHR2, and ARNT2 in dioxin sensitive and -resistant populations of the marine fish Fundulus heteroclitus. AB - Fundulus heteroclitus is a well-characterized marine fish model for studying aryl hydrocarbon toxicity. The F. heteroclitus population in New Bedford Harbor (NBH), a Superfund site in southeastern Massachusetts, exhibits heritable resistance to the toxic effects of planar halogenated aromatic hydrocarbons (PHAHs), including 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD) and polychlorinated biphenyls (PCBs). To investigate the role of the aryl hydrocarbon receptor (AHR) signal transduction pathway in PHAH resistance, we measured the relative levels of AHR1, AHR2, and ARNT2 mRNA in whole embryos at different developmental stages and in dissected tissues of adults, comparing expression of these genes in NBH fish with fish from a reference site (Scorton Creek, MA [SC]). Expression of both AHR1 and AHR2 mRNA increased during development, achieving maximum levels prior to hatching. Maximal embryonic expression of AHR1 was delayed relative to AHR2. Whole NBH and SC embryos exhibited no discernable differences in expression of these genes. As we have previously observed, adult SC fish expressed AHR2 and ARNT2 mRNA in all tissues examined, while AHR1 was expressed predominantly in brain, heart, and gonads. In contrast, AHR1 mRNA was widely expressed in NBH fish, appearing with unusual abundance in gill, gut, kidney, liver, and spleen. This AHR1 expression pattern was not observed in the lab-reared progeny of NBH fish, demonstrating that constitutive AHR1 expression in gill, gut, kidney, liver, and spleen is not a heritable phenotype. Furthermore, widespread AHR1 expression was not induced in reference-site fish by TCDD or PCB mixtures, suggesting that aberrant AHR1 expression is not simply a normal physiological response of contaminant exposure. These results identify ubiquitous AHR1 expression as an attribute unique to feral NBH F. heteroclitus, and they represent a first step in determining the regulatory mechanisms underlying this expression pattern and its possible role in TCDD resistance. PMID- 11006354 TI - Altered hepatic mRNA expression of apoptotic genes during dimethylnitrosamine exposure. AB - The role of TNFalpha in regulating apoptotic signaling was investigated during subacute, low-dose (5.0 mg/kg) dimethylnitrosamine (DMN)-induced hepatotoxicity. In TNFalpha receptor (TNFR) intact (wild-type, WT) mice following 4 and 7 DMN exposures, hepatic transcripts for TNFalpha and TNFR-1 were elevated as compared to vehicle controls. DMN hepatotoxicity in WT and TNFR-1/TNFR-2 double knockout (DKO) mice were then compared over a 7-d exposure period. Liver RNA was isolated to measure hepatic expression of TNFalpha/Fas-related genes and the Bcl-2 family of genes that impact apoptosis. Hepatic mRNA levels for Fas, the apoptosis promoting gene Bax, and the anti-apoptotic gene, Bcl-X(L), were up regulated following 4 and 7 DMN exposures in both WT and TNFR DKO mice as compared to vehicle controls. Notably, hepatic transcript levels for Bax were higher in TNFR DKO mice treated with DMN compared to identically treated WT mice. However, we detected approximately equal DMN-induced apoptotic degradation of liver DNA following 1, 4, and 7 exposures in WT and TNFR DKO mice. Taken together, these data show DMN-induced hepatic TNFalpha expression and suggest that TNFR-1 signaling may be up regulated following 4 and 7 daily DMN exposures. However, TNFalpha is not required for apoptotic signaling at the mRNA transcript level within the liver and instead may actually decrease Bax production. PMID- 11006355 TI - Lack of selective developmental neurotoxicity in rat pups from dams treated by gavage with chlorpyrifos. AB - Pregnant Sprague-Dawley rats were given chlorpyrifos (O:, O-diethyl-O:-[3,5,6 trichloro-2-pyridinyl] phosphorothioate; CPF) in corn oil by gavage from gestation day 6 (GD 6) through lactation day 10 (LD 10) at dosages of 0, 0.3, 1, or 5 mg/kg/day in a developmental neurotoxicity study that conformed to U.S. Environmental Protection Agency 1991 guidelines. GD 0 was the day when evidence of mating was observed and postnatal day 0 (PND 0) was the day of birth. Toxicity was limited to the highest dosage level (5 mg/kg/day) and, in the dams, consisted of muscle fasciculation, hyperpnea, and hyperreactivity. A nonsignificant overall trend toward weight gain and feed consumption was also observed in the high dosage dams, with a statistically significant Group x Time interaction for reduced weight gain in the 5-mg/kg/day group near the end of gestation. Although many developmental indices were normal, pups from high-dosage dams had increased mortality soon after birth, gained weight more slowly than controls, and had several indications of slightly delayed maturation. The early deaths and delayed maturation were attributed to maternal toxicity, though a possible contributing role of direct pup toxicity in delayed development cannot be eliminated. In spite of the apparent delay in physical development, high-dosage pups tested just after weaning had normal learning and memory as tested on a T-maze spatial delayed alternation task. Habituation, a primitive form of learning, was tested in 2 tasks (motor activity and auditory startle) and was not affected. No overt effects were noted in either dams or pups at 1 or 0.3 mg/kg/day. Based on these data, chlorpyrifos produced maternal and developmental toxicity in the 5 mg/kg/day-dosage group. There was no evidence of selective developmental neurotoxicity following exposure to chlorpyrifos. PMID- 11006356 TI - Stage- and species-specific developmental toxicity of all-trans retinoic acid in four native North American ranids and Xenopus laevis. AB - Within the last decade, there have been increasing reports of malformed amphibians across North America. Recently, it has been suggested that hind-limb malformations are a consequence of xenobiotic disruption of developmental pathways regulated by retinoids. To assess the validity of this hypothesis, the developmental toxicity of all-trans retinoic acid (RA) was examined in Xenopus laevis and four North American anurans, at several life stages. To determine the effects of RA on embryogenesis, mid-blastula stage embryos were exposed to 0, 6.25, 12.5, 25, or 50 ng RA/ml for 24 h. To evaluate the effects of RA on hind limb development, early- and mid-limb bud stage tadpoles were exposed to RA concentrations of 0, 250, 500, 750, 1000, or 1250 ng RA/ml for 24 h. Mid-blastula RA exposure resulted in a concentration-dependent increase in dysmorphogenesis and mortality in the three species examined (R. clamitans, R. septentrionalis and X. laevis). RA exposure at stage 51 in X. laevis and stage 28 in R. sylvatica resulted in concentration-dependent increases in reductions and deletions of the hind limb. However, RA was ineffective at inducing hind-limb abnormalities in stages 26 and 28 of R. pipiens, stage 28 in R. clamitans, or stage 48 in X. laevis tadpoles. These results indicate that mid-blastula stage embryos are more sensitive to RA-induced dysmorphogenesis and mortality than limb-bud stage tadpoles. The significance of these findings is discussed in the context of the possible occurrence of retinoid mimics in the environment. PMID- 11006357 TI - Tissue disposition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in maternal and developing long-evans rats following subchronic exposure. AB - Prenatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) produces alterations in the reproductive system of the developing pups. The objective of this study was to determine the disposition of TCDD in maternal and fetal Long Evans (LE) rats following subchronic exposure, since the adverse reproductive and developmental effects have been extensively characterized in this strain of rat. LE rats were dosed by gavage with 1, 10, or 30 ng [(3)H]TCDD/kg in corn oil, 5 days/week for 13 weeks. At the end of 13 weeks, females were mated and dosing continued every day throughout gestation. Dams were sacrificed on gestation day (GD) 9, GD16, GD21, and post-natal day 4 and analyzed for [(3)H]TCDD-derived activity in maternal and fetal tissues. Maternal body burdens were equivalent at different time points, indicating that the dams were at steady state. Maternal body burdens were approximately 19, 120, and 300 ng TCDD/kg following doses of 1, 10, and 30 ng TCDD/kg, respectively. Individual embryo concentrations on GD9 were 1.6, 7, and 16 pg TCDD/g after maternal exposure of 1, 10, and 30 ng/kg/d, respectively. On GD 16, fetal liver, urogenital tract, head, and body concentrations were similar and averaged 1.4, 7.8, and 16.4 pg TCDD/g after administration of 1, 10, or 30 ng TCDD/kg/d, respectively, indicating no preferential sequestration within the different fetal tissues. These concentrations of TCDD within fetal tissues after subchronic exposure are comparable to those seen after a single dose of 50, 200, or 1000 ng TCDD/kg administered on GD15, a critical period of gestation. PMID- 11006358 TI - Evaluation of the developmental toxicity of formamide in Sprague-Dawley (CD) rats. AB - Timed-pregnant CD(R) outbred albino Sprague-Dawley rats received formamide (50, 100, or 200 mg/kg/day) or vehicle (5 ml/kg deionized/distilled water, po) on gestational days (gd) 6 through 19. Maternal food and water consumption (absolute and relative), body weight, and clinical signs were monitored at regular intervals throughout gestation. At termination (gd 20), confirmed-pregnant females (21-23 per group) were evaluated for clinical status and gestational outcome; live fetuses were examined for external, visceral, and skeletal malformations and variations. There were no maternal deaths and no dose-related clinical signs. At 200 mg/kg/day, maternal body weight on gd 20, weight gain, and gravid uterine weight were significantly decreased. Maternal weight gain, corrected for gravid uterine weight, liver weight (absolute or relative), and food and water consumption (absolute or relative), were not affected. Formamide did not affect prenatal viability or incidences of fetal malformations or variations. Average fetal body weight/litter was decreased at 100 and 200 mg/kg/day. Fetal body weight was affected at lower daily doses than in previously published studies, possibly due to the longer total exposure period and/or lack of a recovery period between cessation of exposure and termination. In summary, the maternal toxicity no-observed-adverse-effect level (NOAEL) was 100 mg/kg/day and the low observed adverse effect level (LOAEL) was 200 mg/kg/day under the conditions of this study. Similarly, the developmental toxicity NOAEL was 50 mg/kg/day and the LOAEL was 100 mg/kg/day. PMID- 11006359 TI - Reduced levels of 1,25-dihydroxyvitamin D(3) in rat dams and offspring after exposure to a reconstituted PCB mixture. AB - Previous studies revealed effects of polychlorinated biphenyls (PCBs) and other polyhalogenated hydrocarbons on steroid hormone levels and hormone-dependent functions including behavior. In the present study serum concentrations of the vitamin D(3) metabolites 25-hydroxycholecalciferol (25-D) and 1,25 dihydroxycholecalciferol (1,25-D) were determined in rat dams and offspring after exposure to a PCB mixture that was reconstituted according to the congener pattern found in human breast milk. Unmated females were exposed to diets adulterated with 0; 5; 20; or 40 mg PCBs/kg diet. Exposure started 50 days prior to mating and was terminated at birth. Gestational exposure reduced serum concentrations of 1,25-D in dams in a dose-dependent manner. Concentration of 25 D was also decreased at the time of delivery, but not at weaning. Determination of 1,25-D in offspring at weaning revealed reductions in both high-exposure groups. Levels of 25-D were diminished only at the highest exposure level. Internal PCB concentrations in adipose tissue and brains exhibited a linear relation to dosages in diet. Concentrations of PCBs in brains were similar in dams and offspring at birth, but decreased at the end of lactation in dams. In offspring, values increased during this period because of continued exposure via the milk. In the adipose tissue, PCB levels were much lower in offspring than in dams. To our knowledge, this is the first report of PCB-induced effects on vitamin D(3) metabolites. In dams, reductions were seen even at the lowest exposure level used. Further studies are needed to evaluate the biological significance of these reductions in pregnant dams and possible consequences for the developing offspring. PMID- 11006360 TI - Antiestrogenic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin in mouse uterus: critical role of the aryl hydrocarbon receptor in stromal tissue. AB - The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the role of aryl hydrocarbon receptor (AhR) in estradiol (E(2))-induced uterine epithelial mitogenic activity and secretory protein mRNA expression were determined. Ovariectomized wild-type (wt) and AhR-knockout (AhRKO) mice received oil, E(2), or 5 microg/kg TCDD+E(2). E(2) stimulated similar large increases in the uterine epithelial labeling index (LI) and mRNA abundance for the E(2)-dependent epithelial secretory protein, lactoferrin (LF), in both wt and AhRKO mice. However, uterine epithelial LI and LF mRNA were significantly reduced by TCDD+E(2) in wt but not AhRKO mice. To determine the roles of stromal and epithelial AhR in the TCDD effect, uterine stroma and epithelium from AhRKO and wt mice were enzymatically separated and recombined into four types of tissue recombinants that either contained or lacked AhR in one or more tissue compartments. Tissue recombinants were grafted into nude mice, which were later ovariectomized and given oil, E(2), or TCDD+E(2). Epithelial LI was significantly reduced by TCDD in grafts containing stromal AhR, regardless of epithelial AhR status. However, LI was unaffected by TCDD in grafts lacking stromal AhR, even when epithelial AhR was present. Thus, TCDD inhibits E(2)-induced uterine epithelial mitogenic and secretory activity, and this requires AhR. Anti proliferative effects of TCDD on uterine epithelia appear to be mediated indirectly through stromal AhR, suggesting that liganded AhR alters epithelial function by disrupting normal E(2)-induced stromal activity. This is the first demonstration that TCDD impairs uterine epithelial function by altering normal stromal-epithelial interactions in vivo. PMID- 11006361 TI - Application of a hybrid CFD-PBPK nasal dosimetry model in an inhalation risk assessment: an example with acrylic acid. AB - The available inhalation toxicity information for acrylic acid (AA) suggests that lesions to the nasal cavity, specifically olfactory degeneration, are the most sensitive end point for developing a reference concentration (RfC). Advances in physiologically based pharmacokinetic (PBPK) modeling, specifically the incorporation of computational fluid dynamic (CFD) models, now make it possible to estimate the flux of inhaled chemicals within the nasal cavity of experimental species, specifically rats. The focus of this investigation was to apply an existing CFD-PBPK hybrid model in the estimation of an RfC to determine the impact of incorporation of this new modeling technique into the risk assessment process. Information provided in the literature on the toxicity and mode of action for AA was used to determine the risk assessment approach. A comparison of the approach used for the current U.S. Environmental Protection Agency (U.S. EPA) RfC with the approach using the CFD-PBPK hybrid model was also conducted. The application of the CFD-PBPK hybrid model in a risk assessment for AA resulted in an RfC of 79 ppb, assuming a minute ventilation of 13.8 l/min (20 m(3)/day) in humans. This value differs substantially from the RfC of 0.37 ppb estimated for AA by the U.S. EPA before the PBPK modeling advances became available. The difference in these two RfCs arises from many factors, with the main difference being the species selected (mouse vs. rat). The choice to conduct the evaluation using the rat was based on the availability of dosimetry data in this species. Once these data are available in the mouse, an assessment should be conducted using this information. Additional differences included the methods used for estimating the target tissue concentration, the uncertainty factors (UFs) applied, and the application of duration and uncertainty adjustments to the internal target tissue dose rather than the external exposure concentration. PMID- 11006362 TI - Metabonomics: evaluation of nuclear magnetic resonance (NMR) and pattern recognition technology for rapid in vivo screening of liver and kidney toxicants. AB - The purpose of this study was to evaluate the feasibility of metabonomics technology for developing a rapid-throughput toxicity screen using 2 known hepatotoxicants: carbon tetrachloride (CCl(4)) and alpha-naphthylisothiocyanate (ANIT) and 2 known nephrotoxicants: 2-bromoethylamine (BEA) and 4-aminophenol (PAP). In addition, the diuretic furosemide (FURO) was also studied. Single doses of CCl(4) (0.1 and 0.5 ml/kg), ANIT (10 and 100 mg/kg), BEA (15 and 150 mg/kg), PAP (15 and 150 mg/kg) and FURO (1 and 5 mg) were administered as single IP or oral doses to groups of 4 male Wistar rats/dose. Twenty-four-h urine samples were collected pretest, daily through Day 4, and on Day 10 (high dose CCl(4) and BEA only). Blood samples were taken on Days 1, 2, and 4 or 1, 4, and 10 for clinical chemistry assessment, and the appropriate target organ was examined microscopically. NMR spectra of urine were acquired and the data processed and subjected to principal component analyses (PCA). The results demonstrated that the metabonomic approach could readily distinguish the onset and reversal of toxicity with good agreement between clinical chemistry and PCA data. In at least 2 instances (ANIT and BEA), PCA analysis suggested effects at low doses, which were not as evident by clinical chemistry or microscopic analysis. Furosemide, which had no effect at the doses employed, did not produce any changes in PCA patterns. These data support the contention that the metabonomic approach represents a promising new technology for the development of a rapid throughput in vivo toxicity screen. PMID- 11006363 TI - Peroxisome proliferator-activated receptor alpha-null mice lack resistance to acetaminophen hepatotoxicity following clofibrate exposure. AB - The purpose of this study was to investigate whether activation of the nuclear receptor PPARalpha is needed for protection from acetaminophen (APAP) hepatotoxicity produced by repeated administration of the peroxisome proliferator clofibrate (CFB). Female wild-type and PPARalpha-null mice received corn oil vehicle or 500 mg CFB/kg, ip, daily for 10 days. They were then fasted overnight (18 h) and either killed at 4 or 24 h after challenge with 400 mg APAP/kg. Controls received 50% propylene glycol vehicle only. In this model of CFB hepatoprotection, liver injury was assessed by measuring plasma sorbitol dehydrogenase activity and by histopathology at 24 h after APAP challenge. Significant hepatocellular necrosis was evident in both corn oil-pretreated PPARalpha-null and wild-type mice at 24 h after APAP challenge. In agreement with previous studies, CFB-pretreated wild-type mice showed marked protection against APAP toxicity. In contrast, CFB did not provide protection against APAP hepatotoxicity in the PPARalpha-null mice. Similarly, at 4 h after APAP challenge, hepatic glutathione depletion and selective arylation of cytosolic proteins were reduced significantly in CFB-pretreated wild-type mice, but not in PPARalpha-null mice. The lack of changes in APAP binding and NPSH depletion in CFB-pretreated, PPARalpha-null mice is consistent with the presence of significant liver injury at 24 h in this treatment group. These findings demonstrate that the protection against APAP hepatotoxicity by peroxisome proliferator treatment is mediated by the activation of PPARalpha. PMID- 11006364 TI - CD4(+) T-cell activation and induction of autoimmune hepatitis following trichloroethylene treatment in MRL+/+ mice. AB - Exposure to relatively high levels of trichloroethylene has recently been shown to accelerate the development of an autoimmune response in the autoimmune prone MRL+/+ mice. The trichloroethylene-induced autoimmune response was associated with an increase in activated CD4(+) T cells, producing Th(1)-like cytokines. The present study was conducted to determine whether lower, more occupationally relevant doses of trichloroethylene could also promote autoimmunity, in MRL+/+ mice, and if so, to investigate the mechanism of this accelerated autoimmune response. In addition, histological studies were performed to determine if trichloroethylene was capable of producing pathological markers consistent with an autoimmune disease. Trichloroethylene was administered to mice in the drinking water at 0, 0.1, 0.5, and 2.5 mg/ml for 4 and 32 weeks. There was a significant increase above controls in serum antinuclear antibody (ANA) levels following 4 weeks of both 0.1 and 0.5 mg/kg/day of trichloroethylene. After 32 weeks of treatment, ANA levels were elevated and equal in all groups. The kinetics of the ANA response indicated that trichloroethylene accelerated the innate autoimmune response in the MRL+/+ mice. There was a dose-related increase in the percentage of activated CD4(+) T cells in both the spleens and lymph nodes of mice treated for 32 weeks with trichloroethylene when compared to controls. CD4(+) T cells isolated from MRL+/+ mice after either 4 or 32 weeks of treatment with trichloroethylene secreted inflammatory or Th(1)-like cytokines. Following 32 weeks of trichloroethylene treatment, there was a significant increase in hepatic mononuclear infiltration localized to the portal region, a type of hepatic infiltration consistent with autoimmune hepatitis. Taken collectively, these data suggest that exposure to occupationally relevant concentrations of trichloroethylene can accelerate an autoimmune response and can lead to autoimmune disease. The mechanism of this autoimmunity appears to involve, at least in part, activated CD4(+) T cells that then produced inflammatory cytokines. PMID- 11006365 TI - Intramuscular interferon beta-1a therapy initiated during a first demyelinating event in multiple sclerosis. CHAMPS Study Group. AB - BACKGROUND: Treatment with interferon beta has been shown to help patients with established multiple sclerosis, but it is not known whether initiating treatment at the time of a first clinical demyelinating event is of value. METHODS: We conducted a randomized, double-blind trial of 383 patients who had a first acute clinical demyelinating event (optic neuritis, incomplete transverse myelitis, or brain-stem or cerebellar syndrome) and evidence of prior subclinical demyelination on magnetic resonance imaging (MRI) of the brain. After initial treatment with corticosteroids, 193 patients were randomly assigned to receive weekly intramuscular injections of 30 microg of interferon beta-1a and 190 were assigned to receive weekly injections of placebo. The study end points were the development of clinically definite multiple sclerosis and changes in findings on MRI of the brain. The trial was stopped after a preplanned interim efficacy analysis. RESULTS: During three years of follow-up, the cumulative probability of the development of clinically definite multiple sclerosis was significantly lower in the interferon beta-1a group than in the placebo group (rate ratio, 0.56; 95 percent confidence interval, 0.38 to 0.81; P=0.002). As compared with the patients in the placebo group, patients in the interferon beta-1a group had a relative reduction in the volume of brain lesions (P<0.001), fewer new or enlarging lesions (P<0.001), and fewer gadolinium-enhancing lesions (P<0.001) at 18 months. CONCLUSIONS: Initiating treatment with interferon beta-1a at the time of a first demyelinating event is beneficial for patients with brain lesions on MRI that indicate a high risk of clinically definite multiple sclerosis. PMID- 11006366 TI - Irinotecan plus fluorouracil and leucovorin for metastatic colorectal cancer. Irinotecan Study Group. AB - BACKGROUND: The combination of fluorouracil and leucovorin has until recently been standard therapy for metastatic colorectal cancer. Irinotecan prolongs survival in patients with colorectal cancer that is refractory to treatment with fluorouracil and leucovorin. In a multicenter trial, we compared a combination of irinotecan, fluorouracil and leucovorin with bolus doses of fluorouracil and leucovorin as first-line therapy for metastatic colorectal cancer. A third group of patients received irinotecan alone. METHODS: Patients were randomly assigned to receive irinotecan (125 mg per square meter of body-surface area intravenously), fluorouracil (500 mg per square meter as an intravenous bolus), and leucovorin (20 mg per square meter as an intravenous bolus) weekly for four weeks every six weeks; fluorouracil (425 mg per square meter as an intravenous bolus) and leucovorin (20 mg per square meter as an intravenous bolus) daily for five consecutive days every four weeks; or irinotecan alone (125 mg per square meter intravenously) weekly for four weeks every six weeks. End points included progression-free survival and overall survival. RESULTS: Of 683 patients, 231 were assigned to receive irinotecan, fluorouracil, and leucovorin; 226 to receive fluorouracil and leucovorin; and 226 to receive irinotecan alone. In an intention to-treat analysis, as compared with treatment with fluorouracil and leucovorin, treatment with irinotecan, fluorouracil, and leucovorin resulted in significantly longer progression-free survival (median, 7.0 vs. 4.3 months; P=0.004), a higher rate of confirmed response (39 percent vs. 21 percent, P<0.001), and longer overall survival (median, 14.8 vs. 12.6 months; P=0.04). Results for irinotecan alone were similar to those for fluorouracil and leucovorin. Grade 3 (severe) diarrhea was more common during treatment with irinotecan, fluorouracil, and leucovorin than during treatment with fluorouracil and leucovorin, but the incidence of grade 4 (life-threatening) diarrhea was similar in the two groups (<8 percent). Grade 3 or 4 mucositis, grade 4 neutropenia, and neutropenic fever were less frequent during treatment with irinotecan, fluorouracil, and leucovorin. Adding irinotecan to the regimen of fluorouracil and leucovorin did not compromise the quality of life. CONCLUSIONS: Weekly treatment with irinotecan plus fluorouracil and leucovorin is superior to a widely used regimen of fluorouracil and leucovorin for metastatic colorectal cancer in terms of progression-free survival and overall survival. PMID- 11006367 TI - Multiple complex coronary plaques in patients with acute myocardial infarction. AB - BACKGROUND: Acute myocardial infarction is believed to be caused by rupture of an unstable coronary-artery plaque that appears as a single lesion on angiography. However, plaque instability might be caused by pathophysiologic processes, such as inflammation, that exert adverse effects throughout the coronary vasculature and that therefore result in multiple unstable lesions. METHODS: To document the presence of multiple unstable plaques in patients with acute myocardial infarction and determine their influence on outcome, we analyzed angiograms from 253 patients for complex coronary plaques characterized by thrombus, ulceration, plaque irregularity, and impaired flow. RESULTS: Single complex coronary plaques were identified in 153 patients (60.5 percent) and multiple complex plaques in the other 100 patients (39.5 percent). As compared with patients with single complex plaques, those with multiple complex plaques were less likely to undergo primary angioplasty (86.0 percent vs. 94.8 percent, P = 0.03) and more commonly required urgent bypass surgery (27.0 percent vs. 5.2 percent, P < or = 0.001). During the year after myocardial infarction, the presence of multiple complex plaques was associated with an increased incidence of recurrent acute coronary syndromes (19.0 percent vs. 2.6 percent, P < or = 0.001); repeated angioplasty (32.0 percent vs. 12.4 percent, P < or = 0.001), particularly of non-infarct related lesions (17.0 percent vs. 4.6 percent, P < or = 0.001); and coronary artery bypass graft surgery (35.0 percent vs. 11.1 percent, P < or = 0.001). CONCLUSIONS: Patients with acute myocardial infarction may harbor multiple complex coronary plaques that are associated with adverse clinical outcomes. Plaque instability may be due to a widespread process throughout the coronary vessels, which may have implications for the management of acute ischemic heart disease. PMID- 11006368 TI - Effect of growth hormone treatment on the adult height of children with chronic renal failure. German Study Group for Growth Hormone Treatment in Chronic Renal Failure. AB - BACKGROUND: Growth hormone treatment stimulates growth in short children with chronic renal failure. However, the extent to which this therapy increases final adult height is not known. METHODS: We followed 38 initially prepubertal children with chronic renal failure treated with growth hormone for a mean of 5.3 years until they reached their final adult height. The mean (+/-SD) age at the start of treatment was 10.4+/-2.2 years, the mean bone age was 7.1+/-2.3 years, and the mean height was 3.1+/-1.2 SD below normal. Fifty matched children with chronic renal failure who were not treated with growth hormone served as controls. RESULTS: The children treated with growth hormone had sustained catch-up growth, whereas the control children had progressive growth failure. The mean final height of the growth hormone-treated children was 165 cm for boys and 156 cm for girls. The mean final adult height of the growth hormone-treated children was 1.6+/-1.2 SD below normal, which was 1.4 SD above their standardized height at base line (P< 0.001). In contrast, the final height of the untreated children (2.1+/-1.2 SD below normal) was 0.6 SD below their standardized height at base line (P<0.001). Although prepubertal bone maturation was accelerated in growth hormone-treated children, treatment was not associated with a shortening of the pubertal growth spurt. The total height gain was positively associated with the initial target-height deficit and the duration of growth hormone therapy and was negatively associated with the percentage of the observation period spent receiving dialysis treatment. CONCLUSIONS: Long-term growth hormone treatment of children with chronic renal failure induces persistent catch-up growth, and the majority of patients achieve normal adult height. PMID- 11006369 TI - Liver disease in patients with hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia, or Rendu-Osler-Weber disease, is an autosomal dominant disorder characterized by angiodysplastic lesions (telangiectases and arteriovenous malformations) that affect many organs. Liver involvement in patients with this disease has not been fully characterized. METHODS: We studied the clinical findings and results of hemodynamic, angiographic, and imaging studies in 19 patients with hereditary hemorrhagic telangiectasia and symptomatic liver involvement. RESULTS: We evaluated 14 women and 5 men who ranged in age from 34 to 74 years. All but one of the patients had a hyperdynamic circulation (cardiac index, 4.2 to 7.3 liters per minute per square meter of body-surface area). In eight patients, the clinical findings were consistent with the presence of high-output heart failure. The cardiac index and pulmonary-capillary wedge pressure were elevated in the six patients in whom these measurements were performed. After a median period of 24 months, the condition of three of the eight patients had improved, four were in stable condition with medical therapy, and one had died. Six patients had manifestations of portal hypertension such as ascites or variceal bleeding. The hepatic sinusoidal pressure was elevated in the four patients in whom it was measured. After a median period of 19 months, the condition of two of the six patients had improved, and the other four had died. Five patients had manifestations of biliary disease, such as an elevated alkaline phosphatase level and abnormalities on bile duct imaging. After a median period of 30 months, the condition of two of the five had improved, the condition of one was unchanged, heart failure had developed in one, and one had died after an unsuccessful attempt at liver transplantation. CONCLUSIONS: In patients with hereditary hemorrhagic telangiectasia and symptomatic liver-involvement, the typical clinical presentations include high-output heart failure, portal hypertension, and biliary disease. PMID- 11006370 TI - Images in clinical medicine. Mesenteric ischemia. PMID- 11006371 TI - Multiple sclerosis. PMID- 11006372 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 30-2000. A 25-year-old man with asthma, cardiac failure, diarrhea, and weakness of the right hand. PMID- 11006373 TI - Moving beyond fluorouracil for colorectal cancer. PMID- 11006374 TI - Correction: The Effect of Previous Coronary-Artery Bypass Surgery on the Prognosis of Patients with Diabetes Who Have Acute Myocardial Infarction. PMID- 11006375 TI - Correction: Lack of Effect of a High-Fiber Cereal Supplement on the Recurrence of Colorectal Adenomas. PMID- 11006376 TI - A fibril reinforced nonhomogeneous poroelastic model for articular cartilage: inhomogeneous response in unconfined compression. AB - The depth dependence of material properties of articular cartilage, known as the zonal differences, is incorporated into a nonlinear fibril-reinforced poroelastic model developed previously in order to explore the significance of material heterogeneity in the mechanical behavior of cartilage. The material variations proposed are based on extensive observations. The collagen fibrils are modeled as a distinct constituent which reinforces the other two constituents representing proteoglycans and water. The Young's modulus and Poisson's ratio of the drained nonfibrillar matrix are so determined that the aggregate compressive modulus for confined geometry fits the experimental data. Three nonlinear factors are considered, i.e. the effect of finite deformation, the dependence of permeability on dilatation and the fibril stiffening with its tensile strain. Solutions are extracted using a finite element procedure to simulate unconfined compression tests. The features of the model are then demonstrated with an emphasis on the results obtainable only with a nonhomogeneous model, showing reasonable agreement with experiments. The model suggests mechanical behaviors significantly different from those revealed by homogeneous models: not only the depth variations of the strains which are expected by qualitative analyses, but also, for instance, the relaxation-time dependence of the axial strain which is normally not expected in a relaxation test. Therefore, such a nonhomogeneous model is necessary for better understanding of the mechanical behavior of cartilage. PMID- 11006377 TI - Tensegrity architecture explains linear stiffening and predicts softening of living cells. AB - The problem of theoretical explanation of the experimentally observed linear stiffening of living cells is addressed. This explanation is based on Ingber's assumption that the cell cytoskeleton, which enjoys tensegrity architecture with compressed microtubules that provide tension to the microfilaments, affects the mechanical behavior of the living cell. Moreover, it is shown that the consideration of the extreme flexibility of microtubules and the unilateral response of microfilaments is crucial for the understanding of the living cell overall behavior. Formal nonlinear structural analysis of the cell cytoskeleton under external mechanical loads is performed. For this purpose, a general computer model for tensegrity assemblies with unilateral microfilaments and buckled microtubules is developed and applied to the theoretical analysis of the mechanical response of 2D and 3D examples of tensegrity cells mimicking the behavior of real living cells. Results of the computer simulations explain the experimentally observed cell stiffening. Moreover, the theoretical results predict the possible existence of a transient softening behavior of cells, a phenomenon, which has not been observed in experiments yet. PMID- 11006378 TI - Knoop microhardness anisotropy of the ovine radius. AB - The Knoop indenter has been used to characterise fully the Knoop microhardness (H(K)) anisotropy of compact bone. 2120 indentations were performed on mature ovine radii and a linear relationship was found between H(K) and the angle between the major diagonal of the indenter and the lamella boundaries (p<<0.001). H(K) increased significantly with ash fraction (p<0.001), but decreased with atmospheric vapour pressure (p<0.05). A significant interaction was found between ash fraction and atmospheric vapour pressure (p<0.01). H(K) significantly varied with indentation position along the diaphysis and around the cortex (both p<<0.001), however radial variation in H(K) was not statistically significant. The variation of ash fraction showed similar trends. These data show that H(K) varies similarly to Vickers microhardness, but in addition, can provide clear information on the anisotropy of Haversian bone without the need for excising many different indentation planes. A large number of indentations are required to obtain low type I and type II errors in the statistical analysis. PMID- 11006379 TI - In vitro axial preload application during spine flexibility testing: towards reduced apparatus-related artefacts. AB - Presently, there is little consensus about how, or even if, axial preload should be incorporated in spine flexibility tests in order to simulate the compressive loads naturally present in vivo. Some preload application methods are suspected of producing unwanted "artefact" forces as the specimen rotates and, in doing so, influencing the resulting kinematics. The objective of this study was to quantitatively compare four distinct types of preload which have roots in contemporary experimental practice. The specific quantities compared were the reaction moments and forces resulting at the intervertebral disc and specimen kinematics. The preload types incorporated increasing amounts of caudal constraint on the preload application vector ranging from an unconstrained dead load arrangement to an apparatus that allowed the vector to follow rotations of the specimen. Six human cadaveric spine segments were tested (1-L1/L2, 3-L2/L3, 1 L3/L4 and 1-L4/L5). Pure moments were applied to the specimens with each of the four different types of compressive preload. Kinematic response was measured using an opto-electronic motion analysis system. A six-axis load cell was used to measure reaction forces and moments. Artefact reaction moments and shear forces were significantly affected by preload application method and magnitude. Unconstrained preload methods produced high artefact moments and low artefact shear forces while more constrained methods did the opposite. A mechanical trade off is suggested by our results, whereby unwanted moment can only be prevented at the cost of shear force production. When comparing spine flexibility studies, caution should be exercised to ensure preload was applied in a similar manner for all studies. Unwanted moments or forces induced as a result of preload application method may render the comparison of two seemingly similar studies inappropriate. PMID- 11006380 TI - Collagen fibril D-period may change as a function of strain and location in ligament. AB - The purpose of this study was to determine if the characteristic banding pattern (D-period) of collagen fibrils from rabbit medial collateral ligaments changes as a function of gross ligament strain and, if so, whether the changes are location dependent (insertion versus midsubstance). Femur-medial collateral ligament-tibia complexes were strained to 0, 8, or 12% and immediately chemically fixed in situ. Samples were taken from the medial collateral ligament midsubstance and bony insertions, and prepared for and observed under a transmission electron microscope. D-period length was measured and found to increase (albeit not significantly so, p=0. 1) as a function of gross strain for samples obtained from the insertion sites but not for samples obtained from the ligament midsubstance. Results suggested that ligament strains are inhomogeneous at the ultrastructural level. PMID- 11006381 TI - High-resolution finite element models with tissue strength asymmetry accurately predict failure of trabecular bone. AB - The ability to predict trabecular failure using microstructure-based computational models would greatly facilitate study of trabecular structure function relations, multiaxial strength, and tissue remodeling. We hypothesized that high-resolution finite element models of trabecular bone that include cortical-like strength asymmetry at the tissue level, could predict apparent level failure of trabecular bone for multiple loading modes. A bilinear constitutive model with asymmetric tissue yield strains in tension and compression was applied to simulate failure in high-resolution finite element models of seven bovine tibial specimens. Tissue modulus was reduced by 95% when tissue principal strains exceeded the tissue yield strains. Linear models were first calibrated for effective tissue modulus against specimen-specific experimental measures of apparent modulus, producing effective tissue moduli of (mean+/-S.D.) 18.7+/-3.4GPa. Next, a parameter study was performed on a single specimen to estimate the tissue level tensile and compressive yield strains. These values, 0.60% strain in tension and 1.01% strain in compression, were then used in non-linear analyses of all seven specimens to predict failure for apparent tensile, compressive, and shear loading. When compared to apparent yield properties previously measured for the same type of bone, the model predictions of both the stresses and strains at failure were not statistically different for any loading case (p>0.15). Use of symmetric tissue strengths could not match the experimental data. These findings establish that, once effective tissue modulus is calibrated and uniform but asymmetric tissue failure strains are used, the resulting models can capture the apparent strength behavior to an outstanding level of accuracy. As such, these computational models have reached a level of fidelity that qualifies them as surrogates for destructive mechanical testing of real specimens. PMID- 11006382 TI - Serum modulates the intracellular calcium response of primary cultured bone cells to shear flow. AB - We investigated the effect of newborn bovine serum on the intracellular calcium [Ca(2+)](i) response of primary cultured bone cells stimulated by fluid flow. As it has been previously established that these cells exhibit [Ca(2+)](i) responses to fluid flow shear stress in saline media without growth factors or other chemically stimulatory factors, we hypothesized that the addition of serum to the flow medium would enhance the mechanosensitivity of the cells. We examined the effect of a short-term (10-15min) exposure of the cells to 2 and 10% serum prior to flow stimulation (pretreated) compared to not exposing the cells prior to flow stimulation (unpretreated). The cells were subjected to a well-defined, 90-s flow stimulus with shear stress levels ranging from 0.02 to 3.5Pa in a laminar flow chamber using a saline medium supplemented with 2 or 10% serum. For pretreatment, the serum concentration was the same from pre-flow to flow exposure. We observed a differential effect in the magnitude of the peak [Ca(2+)](i) response modulated by the concentration of serum in the pre-flow medium. Additionally, ATP supplemented flow was examined as a comparison to the serum-supplemented flow and exhibited a similar trend in the peak [Ca(2+)](i) flow response that was dependent on ATP concentration and pre-flow exposure conditions. These findings demonstrate that under the conditions of this study, chemical agonist exposure can modulate the [Ca(2+)](i) response in bone cells subjected to fluid flow induced shear stress. PMID- 11006383 TI - Digital automated RSA compared to manually operated RSA. AB - The accuracy of digital Roentgen stereophotogrammetric analysis (RSA) was compared to the accuracy of a manually operated RSA system. For this purpose, we used radiographs of a phantom and radiographs of patients. The radiographs of the patients consisted of double examinations of 12 patients that had a tibial osteotomy and of double examinations of 12 patients that received a total hip prosthesis. First, the radiographs were measured manually with an accurate measurement table. Subsequently, the images were digitized by a film scanner at 150 DPI and 300 DPI resolutions and analyzed with the RSA-CMS software. In the phantom experiment, the manually operated system produced significantly better results than the digital system, although the maximum difference between the median values of the manually operated system and the digital system was as low as 0.013mm for translations and 0.033 degrees for rotations. In the radiographs of the patients, the manually operated system and the digital system produced equally accurate results: no significant differences in translations and rotations were found. We conclude that digital RSA is an accurate, fast, and user friendly alternative for manually operated RSA. Currently, digital RSA systems are being used in a growing number of clinical RSA-studies. PMID- 11006384 TI - Quantification of fingertip force reduction in the forefinger following simulated paralysis of extensor and intrinsic muscles. AB - Objective estimates of fingertip force reduction following peripheral nerve injuries would assist clinicians in setting realistic expectations for rehabilitating strength of grasp. We quantified the reduction in fingertip force that can be biomechanically attributed to paralysis of the groups of muscles associated with low radial and ulnar palsies. We mounted 11 fresh cadaveric hands (5 right, 6 left) on a frame, placed their forefingers in a functional posture (neutral abduction, 45 degrees of flexion at the metacarpophalangeal and proximal interphalangeal joints, and 10 degrees at the distal interphalangeal joint) and pinned the distal phalanx to a six-axis dynamometer. We pulled on individual tendons with tensions up to 25% of maximal isometric force of their associated muscle and measured fingertip force and torque output. Based on these measurements, we predicted the optimal combination of tendon tensions that maximized palmar force (analogous to tip pinch force, directed perpendicularly from the midpoint of the distal phalanx, in the plane of finger flexion extension) for three cases: non-paretic (all muscles of forefinger available), low radial palsy (extrinsic extensor muscles unavailable) and low ulnar palsy (intrinsic muscles unavailable). We then applied these combinations of tension to the cadaveric tendons and measured fingertip output. Measured palmar forces were within 2% and 5 degrees of the predicted magnitude and direction, respectively, suggesting tendon tensions superimpose linearly in spite of the complexity of the extensor mechanism. Maximal palmar forces for ulnar and radial palsies were 43 and 85% of non-paretic magnitude, respectively (p<0.05). Thus, the reduction in tip pinch strength seen clinically in low radial palsy may be partly due to loss of the biomechanical contribution of forefinger extrinsic extensor muscles to palmar force. Fingertip forces in low ulnar palsy were 9 degrees further from the desired palmar direction than the non-paretic or low radial palsy cases (p<0.05). PMID- 11006385 TI - Large-sliding contact elements accurately predict levels of bone-implant micromotion relevant to osseointegration. AB - Primary stability is recognised as an important determinant in the aseptic loosening failure process of cementless implants. An accurate evaluation of the bone-implant relative micromotion is becoming important both in pre-clinical and clinical studies. If the biological threshold for micro-movements is in the range 100-200 micrometer then, in order to be discriminative, any method used to evaluate the primary stability should have an accuracy of 10-20 micrometer or better. Additionally, such method should also be able to report the relative micromotion at each point of the interface. None of the available experimental methods satisfies both requirements. Aim of the present study is to verify if any of the current finite element modelling techniques is sufficiently accurate in predicting the primary stability of a cementless prosthesis to be used to decide whether the micromotion may or may not jeopardise the implant osseointegration. The primary stability of an anatomic cementless stem, as measured in vitro, was used as a benchmark problem to comparatively evaluate different contact modelling techniques. Frictionless contact, frictional contact and press-fitted frictional contact conditions were modelled using alternatively node-to-node, node-to-face and face-to-face contact elements. The model based on face-to-face contact elements accounting for frictional contact and initial press-fit was able to predict the micromotion measured experimentally with an average (RMS) error of 10 micrometer and a peak error of 14 micrometer. All the other models presented errors higher than 20 micrometer assumed in the present study as an accuracy threshold. PMID- 11006386 TI - Predicted region of stability for balance recovery: motion at the knee joint can improve termination of forward movement. AB - Earlier experimental studies on balance recovery following perturbation have identified two discrete strategies commonly employed by humans, i.e. hip and ankle strategies. It has hence been implied that the knee joint plays a relatively minor role in balance recovery. The purpose of this study was to determine whether the size of the feasible stability region (FSR) would be affected by allowing knee motion in sagittal plane movement termination. The FSR was defined as the feasible range of anterior velocities of the center of mass (COM) of a human subject that could be reduced to zero with the final COM position within the base of support (BOS) limits. The FSR was computed using a four-segment biomechanical model and optimization routine based on Simulated Annealing algorithm for three scenarios: unrestricted knee motion (UK), restricted knee motion (RK), and unrestricted knee motion with an initial posture that matches RK (UKM). We found that movement termination could benefit little from UK condition when the COM (x(COM)) was initially located in the forefoot region [0.00 (toe) >x(COM)>/=-0.50 (mid-foot)] with no more than a 17% increase in FSR compared to RK. The effect of knee motion increased in the rear foot region with a 25% increase in FSR at x(COM)=-1 (heel). Close to half of this difference (12%) was attributable to the knee-related restriction on initial posture and the rest to movement termination per se. These findings illustrated a theoretical role of knee motion in standing humans' repertoire of effective posture responses, which include hip and ankle strategies and their variants for balance recovery with stationary BOS. PMID- 11006387 TI - The glenohumeral joint rotation centre in vivo. AB - Within the framework of the current call for standardization in upper extremity research, three methods to determine the glenohumeral joint rotation centre in vivo were tested. Therefore, subjects performed humeral movements, while a 3D electromagnetic tracking device registered the motion of the humerus with respect to the scapula. For the first method to estimate the glenohumeral joint rotation centre five scapular bony landmarks served as input to regression equations. The second method fitted a sphere through the humeral position data and the third method calculated the rotation centre determining an optimal helical axis. The experiment consisted of two parts, at first one subject was measured 10 times, subsequently one observer measured 10 subjects twice and another observer measured these subjects once. The first part of the experiment demonstrated that all methods are capable to reproduce the rotation centre within 4mm, but the location of the centre differed significantly between methods (p<0.001). The second part, showed that inter- and intra-observer reliability was sufficiently for the sphere-fitting method and for the helical-axes method. The two observations of one observer differed significantly (p<0.008) using the regression method. The authors prefer the helical-axes method, it is a reliable and valid method which can be applied in movement registration of healthy subjects and patients with a shoulder endoprosthesis, it can be applied in hinge joints to determine a rotation axis instead of a rotation centre which is desirable in standardized upper extremity research, and calculation time is short. PMID- 11006388 TI - Numerical 3D analysis of oscillatory flow in the time-varying laryngeal channel. AB - Three-dimensional flow through an anatomically representative model of the human larynx has been numerically simulated. This model includes the vestibular folds, the vocal cords and the glottic and subglottic areas. Pseudo-time-varying glottic aperture and flow conditions have been considered during quiet breathing, with a peak volume flow rate of 0.75l/s and a frequency of 0.25Hz. Because of the severe constriction, jet-like configurations have been observed. Minor differences have been outlined between the inspiration and expiration profiles. Simulations demonstrated the presence of a backflow region which may extend to 60mm from the glottis at peak inspiration and occupy 20% of the tracheal cross section. Because of its rolling, this backflow region appears in the sagittal plane close to the anterior wall, only one diameter from the laryngeal constriction and extends over about 40mm. The evolution of the streamwise velocity contours and of the corresponding secondary vector plots at six critical stations, including the glottic section, has also been described. A double pair of counter-rotating vortices develops shortly downstream/upstream from the orifice respectively at inspiration/expiration and merges near the frontal plane about 25mm from the glottis. The effect of the incoming flow has been evaluated by including the pharyngeal channel; no major difference has been observed in the computed flow patterns. PMID- 11006389 TI - Acetabular morphology and resurfacing design. AB - The bony surfaces of 18 archaeological hemipelves were scanned using a 3D laser surface scanner and CyDir software on a Silicon Graphics workstation. The acetabular area was selected and point data from the approximately spherical bone surface saved. These data were input to a MATLAB routine that calculated the radius and centre of the best-fit sphere. The goodness of fit was estimated using the mean and standard deviation of the distance of the bone surface points from the sphere surface. Eight points, at approximately equal distances around the acetabular rim, were selected with reference to bony landmarks. A plane containing three of these points served as an orientation reference plane. The vectors joining the eight rim points to the centre of the best-fit sphere were found. The angles between these vectors and the normal to the reference plane were calculated. Paired angles were summed to give the angle subtended by the acetabular rim in four directions. The overall mean angle was 158 degrees (range of mean angles 145 degrees -173 degrees ). The largest individual angles, some exceeding 180 degrees, were in the superior-inferior direction, while the mean angle in the anterior-posterior direction, i.e. that controlling flexion extension, was 152 degrees. Males had larger subtended angles than females, although the difference was not statistically significant. Simulated reaming increased all angles by approximately 10 degrees. The subtended angles are important parameters in the design of the acetabular component of a hip replacement and particularly important in resurfacing hip replacement when the volume available is tightly constrained. PMID- 11006390 TI - Analysis of a femoral hip prosthesis designed to reduce stress shielding. AB - The natural stress distribution in the femur is significantly altered after total hip arthroplasty (THA). When an implant is introduced, it will carry a portion of the load, causing a reduction of stress in some regions of the remaining bone. This phenomenon is commonly known as stress shielding. In response to the changed mechanical environment the shielded bone will remodel according to Wolff's law, resulting in a loss of bone mass through the biological process called resorption. Resorption can, in turn, cause or contribute to loosening of the prosthesis. The problem is particularly common among younger THA recipients. This study explores the hypothesis that through redesign, a total hip prosthesis can be developed to substantially reduce stress shielding. First, we describe the development of a new femoral hip prosthesis designed to alleviate this problem through a new geometry and system of proximal fixation. A numerical comparison with a conventional intramedullary prosthesis as well as another proximally fixed prosthesis, recently developed by Munting and Verhelpen (1995. Journal of Biomechanics 28(8), 949-961) is presented. The results show that the new design produces a more physiological stress state in the proximal femur. PMID- 11006391 TI - The mechanical environment of the chondrocyte: a biphasic finite element model of cell-matrix interactions in articular cartilage. AB - Mechanical compression of the cartilage extracellular matrix has a significant effect on the metabolic activity of the chondrocytes. However, the relationship between the stress-strain and fluid-flow fields at the macroscopic "tissue" level and those at the microscopic "cellular" level are not fully understood. Based on the existing experimental data on the deformation behavior and biomechanical properties of articular cartilage and chondrocytes, a multi-scale biphasic finite element model was developed of the chondrocyte as a spheroidal inclusion embedded within the extracellular matrix of a cartilage explant. The mechanical environment at the cellular level was found to be time-varying and inhomogeneous, and the large difference ( approximately 3 orders of magnitude) in the elastic properties of the chondrocyte and those of the extracellular matrix results in stress concentrations at the cell-matrix border and a nearly two-fold increase in strain and dilatation (volume change) at the cellular level, as compared to the macroscopic level. The presence of a narrow "pericellular matrix" with different properties than that of the chondrocyte or extracellular matrix significantly altered the principal stress and strain magnitudes within the chondrocyte, suggesting a functional biomechanical role for the pericellular matrix. These findings suggest that even under simple compressive loading conditions, chondrocytes are subjected to a complex local mechanical environment consisting of tension, compression, shear, and fluid pressure. Knowledge of the local stress and strain fields in the extracellular matrix is an important step in the interpretation of studies of mechanical signal transduction in cartilage explant culture models. PMID- 11006392 TI - Intrinsic and extrinsic contributions to the passive moment at the metacarpophalangeal joint. AB - The purpose of this investigation was to determine whether the passive range of motion at the finger joints is restricted more by intrinsic tissues (cross a single joint) or by extrinsic tissues (cross multiple joints). The passive moment at the metacarpophalangeal (MP) joint of the index finger was modeled as the sum of intrinsic and extrinsic components. The intrinsic component was modeled only as a function of MP joint angle. The extrinsic component was modeled as a function of MP joint angle and wrist angle. With the wrist fixed in seven different positions the passive moment at the MP joint of eight subjects was recorded as the finger was rotated through its range at a constant rate. The moment-angle data were fit by the model and the extrinsic and intrinsic components were calculated for a range of MP joint angles and wrist positions. With the MP joint near its extension limit, the median percent extrinsic contribution was 94% with the wrist extended 60 degrees and 14% with the wrist flexed 60 degrees. These percentages were 40 and 88%, respectively, with the MP joint near its flexion limit. Our findings indicate that at most wrist angles the extrinsic tissues offer greater restraint at the limits of MP joint extension and flexion than the intrinsic tissues. The intrinsic tissues predominate when the wrist is flexed or extended enough to slacken the extrinsic tissues. Additional characteristics of intrinsic and extrinsic tissues can be deduced by examining the parameter values calculated by the model. PMID- 11006393 TI - Mechanical properties of compacted morselized cancellous bone graft using one dimensional consolidation testing. AB - Failures of orthopaedic procedures that use morselized cancellous bone (MCB) graft for load bearing are often due to gross displacement within the graft material. For this reason the mechanical behavior of MCB must be better understood. Our purpose is to present a detailed testing methodology for the mechanical characterization of MCB, and to illustrate how this methodology can be used to study the influence of water and fat content. Complete one-dimensional consolidation testing was performed on bovine cancellous bone processed to represent MCB typically used in surgery (52% water, 31% fat). The one-dimensional consolidation strain under a stress of 1.09MPa was 30.9% and the confined modulus was 8.0MPa. The coefficient of consolidation (rate of consolidation) was 2. 2x10( 5)cm(2)/s and the coefficient of secondary strain (steady-state creep rate) was 1.9%. While reducing the water content alone had some influence on properties, reducing the fat content improved both the static and dynamic behavior. A sample of MCB which had fat intentionally minimized and a lower overall moisture content (56% water, 5% fat) demonstrated 23.1% strain, a confined modulus of 9.6MPa, a coefficient of consolidation of 3.4x10(-3)cm(2)/s, and a coefficient of secondary strain of 0.9%. The test methods described in this technical note can be used to evaluate the influence of fluid content on the mechanical behavior of MCB. PMID- 11006394 TI - Chronic softening of cartilage without thickening of underlying bone in a joint trauma model. AB - We have recently developed a trauma model to study degradation of the rabbit patello-femoral joint. Our current working hypothesis is that alterations in retropatellar cartilage and underlying bone in our model are initiated independently by acute overstresses developed in each tissue during blunt insult to the joint, and that the processes of chronic degradation in each tissue are not related in a mechanical sense. The current study was conducted in an attempt to help validate our hypothesis by impacting the patello-femoral joint with a padded interface. Based upon earlier human cadaver experiments, we believe this would reduce the acute overstresses in patellar bone while the stresses developed in the overlying retropatellar cartilage would be sufficient enough to initiate a chronic softening of the tissue. Twenty-four animals received an impact to the patello-femoral joint and were sacrificed at either 0, 4.5, or 12 months post insult. Three acute animals were impacted to develop a simplified computational model to estimate the stresses in joint tissues. The study showed there was a significant softening of the retropatellar cartilage at 4.5 and 12 months post trauma, compared to unimpacted controls. However, no thickening of the underlying subchondral bone was documented at any timepoint. This was consistent with a reduction of stress in the bone compared to earlier studies, which document thickened subchondral bone post-insult at the same applied impact load. In conclusion, this study helped validate our hypothesis by documenting chronic softening of cartilage without remodeling of the underlying subchondral bone. Furthermore, this study, along with our earlier studies, suggest that impact load alone, which is currently used by the automobile industry to certify new automobiles, is not a good predictor of chronic injuries to a diarthrodial joint, and that simply the addition of padding to impact interfaces may not be adequate to protect occupants from chronic injuries. PMID- 11006395 TI - Comparison of different calculations of three-dimensional joint kinematics from video-based system data. AB - When skin-fixed marker trajectories are used to calculate 3D joint kinematics, the measurement errors (i.e. the difference between the trajectories of the external markers and those of the skeleton) influence to some extent the accuracy of the results, depending both on the calculation method and on the axes about which the rotations are expressed. The purpose of this paper is to compare several expressions of joint angular variations. Two kinematic concepts are used to calculate the changes in the orientation of the distal segment versus the proximal one: the first method consists of computing the components of the spatial attitude vector, the second one deals with the determination of elementary rotations about successive axes. For each of these methods, two sets of three axes are tested to express the results: the axes forming the reference frame affixed to the body segment adjacent to the joint (named fixed axes), and a set consisting of a first axis belonging to the proximal segment, a third axis belonging to the distal segment and a second (floating) axis defined as the cross product between the two other ones (named mobile axes). To compare these four distinct expressions on the knee joint, numerical simulations of perturbed skin marker trajectories are performed, based on experimental data recorded by a Motion Analysis system during a normal gait cycle. A significant difference is pointed out only for the internal-external rotation angle, for which the best expression - from the viewpoint of sensitivity to experimental errors - is obtained using the components of the attitude vector in a segment-embedded reference frame. PMID- 11006396 TI - An empirical approach to characterizing trunk muscle coactivation using simulation input modeling techniques. AB - Accurately describing trunk muscle coactivation is fundamental to quantifying the spine reaction forces that occur during lifting tasks and has been the focus of a great deal of research in the spine biomechanics literature. One limitation of previous approaches has been a lack of consideration given to the variability in these coactivation strategies. The research presented in this paper is an empirical approach to quantifying and modeling trunk muscle coactivation using simulation input modeling techniques. Electromyographic (EMG) data were collected from 28 human subjects as they performed controlled trunk extension exertions. These exertions included isokinetic (10 and 45 degrees /s) and constant acceleration (50 degrees /s/s) trunk extensions in symmetric and asymmetric (30 degrees ) postures at two levels of trunk extension moment (30 and 80Nm). The EMG data were collected from the right and left pairs of the erector spinae, latissimus dorsi, rectus abdominis, external obliques and internal obliques. Each subject performed nine repetitions of each combination of independent variables. The data collected during these trials were used to develop marginal distributions of trunk muscle activity as well as a 10x10 correlation matrix that described how the muscles cooperated to produce these extension torques. These elements were then combined to generate multivariate distributions describing the coactivation of the trunk musculature. An analysis of these distributions revealed that increases in extension moment, extension velocity and sagittal flexion angle created increases in both the mean and the variance of the distributions of the muscular response, while increases in the rate of trunk extension acceleration decreased both the mean and variance of the distributions of activity across all muscles considered. Increases in trunk asymmetry created a decrease in mean of the ipsi-lateral erector spinae and an increase in the mean of all other muscles considered, but there was little change in the variance of these distributions as a function of asymmetry. PMID- 11006397 TI - A method for measuring endpoint stiffness during multi-joint arm movements. AB - Current methods for measuring stiffness during human arm movements are either limited to one-joint motions, or lead to systematic errors. The technique presented here enables a simple, accurate and unbiased measurement of endpoint stiffness during multi-joint movements. Using a computer-controlled mechanical interface, the hand is displaced relative to a prediction of the undisturbed trajectory. Stiffness is then computed as the ratio of restoring force to displacement amplitude. Because of the accuracy of the prediction (< 1 cm error after 200 ms) and the quality of the implementation, the movement is not disrupted by the perturbation. This technique requires only 13 as many trials to identify stiffness as the method of Gomi and Kawato (1997, Biological Cybernetics 76, 163-171) and may, therefore, be used to investigate the evolution of stiffness during motor adaptation. PMID- 11006398 TI - The position of the rotation center of the glenohumeral joint. AB - To validate the assumption that the center of rotation in the glenohumeral (GH) joint can be described based on the geometry of the joint, two methods for calculation of the GH rotation center were compared. These are a kinematic estimation based on the calculation of instantaneous helical axes, and a geometric estimation based on a spherical fit through the surface of the glenoid. Four fresh cadaver arms were fixed at the scapula and fitted with electromagnetic sensors. Each arm was moved in different directions while at the same time the orientation of the humerus was recorded. Subsequently, each specimen was dissected and its glenoid and humeral head surfaces were digitized. Results indicate no differences between the methods. It is concluded that the method to estimate the GH center of rotation as the center of a sphere through the glenoid surface, with the radius of the humeral head, appears to be valid. PMID- 11006399 TI - High-precision satellite positioning system as a new tool to study the biomechanics of human locomotion. AB - New Global Positioning System (GPS) receivers allow now to measure a location on earth at high frequency (5Hz) with a centimetric precision using phase differential positioning method. We studied whether such technique was accurate enough to retrieve basic parameters of human locomotion. Eight subjects walked on an athletics track at four different imposed step frequencies (70-130steps/min) plus a run at free pace. Differential carrier phase localization between a fixed base station and the mobile antenna mounted on the walking person was calculated. In parallel, a triaxial accelerometer, attached to the low back, recorded body accelerations. The different parameters were averaged for 150 consecutive steps of each run for each subject (total of 6000 steps analyzed). We observed a perfect correlation between average step duration measured by accelerometer and by GPS (r=0.9998, N=40). Two important parameters for the calculation of the external work of walking were also analyzed, namely the vertical lift of the trunk and the velocity variation per step. For an average walking speed of 4.0km/h, average vertical lift and velocity variation were, respectively, 4.8cm and 0.60km/h. The average intra-individual step-to-step variability at a constant speed, which includes GPS errors and the biological gait style variation, were found to be 24. 5% (coefficient of variation) for vertical lift and 44.5% for velocity variation. It is concluded that GPS technique can provide useful biomechanical parameters for the analysis of an unlimited number of strides in an unconstrained free-living environment. PMID- 11006400 TI - Hysteresis measurements in intact human tendon. AB - Mechanical hysteresis in tendons has traditionally been quantified from tensile testing of isolated specimens. Limitations associated with tendon displacement measurement and clamping, and uncertainties as to whether in vitro material represents intact tendon function necessitate measuring hysteresis under in vivo conditions. In the present study such measurements were taken in the human tibialis anterior (TA) tendon. Having the foot fixed on a dynamometer footplate, the displacement of the TA tendon during stimulation and relaxation of the TA muscle was recorded by means of ultrasonography in six men. Combining moment data corresponding to 0, 20, 40, 60, 80 and 100% of maximum voltage moment and the respective tendon-displacement data, a hysteresis loop was obtained between the load-displacement curves during contraction and relaxation. Measurement of the hysteresis loop area yielded a value of 19%. This value agrees with results from in vitro tensile tests of low-stress tendons, suitable for tensile force transmission and joint displacement control. In fact, the human TA tendon has such functional characteristics. The methodology presented allows design of longitudinal and cross-sectional experimental protocols, and in vivo assessment of tendon function and propensity to overheat. PMID- 11006401 TI - On the independence of time and strain effects in the stress relaxation of ligaments and tendons. AB - The hypothesis of variables separation, namely the time and the strain separation in the relaxation function, is widely used in soft tissue biomechanics. Although this hypothesis is central to several biomechanical models, only few experimental works have tried to verify it. From these studies, contradictory results have been found. Moreover, it has recently been noted that no such experimental verification has been performed for ligament tissues. In this paper, an experimental method is developed to test the hypothesis of variables separation. This method is then used with human cruciate ligaments and patellar tendons. It is shown that the use of the variables separation hypothesis is justified at least for strain values lower than 16% for anterior cruciate ligament, lower than 12% for posterior cruciate ligament and lower than 6% for patellar tendon. The method presented in this paper could be used to verify the validity of variables separation for other tissues. PMID- 11006402 TI - Postural control of trunk during unstable sitting. AB - A method for quantifying postural control of the lumbar spine during unstable sitting was developed. The unstable seat apparatus was equipped with leg and foot supports to isolate the control of the lumbar spine and trunk from the adjustments in the lower body joints. Polyester resin hemispheres with decreasing diameters were attached to the bottom of the seat to achieve increasing levels of task difficulty. The seat was placed on a force plate at the edge of a table and the participating subjects were instructed to maintain their balance while sitting on the seat. Coordinates of center of pressure (CoP) were recorded and quantified with summary statistics and random walk analysis. The CoP movement increased significantly with increased seat instability (task difficulty) (p<0.01). Stabilogram plots of the CoP movement revealed short and long-term regions consistent with the hypothesis that the two regions reflect open and closed-loop postural control mechanisms. Repeatability of the CoP parameters was excellent for the summary statistics and the short-term random walk coefficients (0.77 or = 4-fold over vehicle control, in all studies. These data demonstrate the reproducibility of positive control response in MN assay in our laboratory and support the MN Assay Expert Panel's view that the use of positive control may not be necessary in every study. PMID- 11006412 TI - Mutation spectra in Salmonella of analogues of MX: implications of chemical structure for mutational mechanisms. AB - We determined the mutation spectra in Salmonella of four chlorinated butenoic acid analogues (BA-1 through BA-4) of the drinking water mutagen 3-chloro-4 (dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) and compared the results with those generated previously by us for MX and a related compound, MCF. We then considered relationships between the properties of mutagenic potency and mutational specificity for these six chlorinated butenoic acid analogues. In TA98, the three most potent mutagens, BA-3, BA-4, MX, and the organic extract, all induced large percentages of complex frameshifts (33-67%), which distinguish these agents from any other class of compound studied previously. In TA100, which has only GC sites for mutation recovery, >71% of the mutations induced by all of the agents were GC-->TA transversions. The availability of both GC and TA sites for mutation in TA104 resulted in greater distinctions in mutational specificity than in TA100. MX targeted GC sites almost exclusively (98%); the structurally similar BA-4 and BA-2 produced mutations at similar frequencies at both GC and AT sites; and the structurally similar BA-3 and BA-1 induced most mutations at AT sites (69%). Thus, large variations in structural properties influencing relative mutagenic potency appeared to be distinct from the more localized similar structural features influencing mutagenic specificity in TA104. Among a set of physicochemical properties examined for the six butenoic acids, a significant correlation was found between pK(a) and mutagenic potency in TA100, even when the unionized fraction of the activity dose was considered. In addition, a correlation in CLOGP for BA-1 to BA-4 suggested a role for bioavailability in determining mutagenic potency. These results illustrate the potential value of structural analyses for exploring the relationship between chemical structure and mutational mechanisms. To our knowledge, this is the first study in which such analyses have been applied to structural analogues for which both mutagenic potency and mutation spectra date were available. PMID- 11006413 TI - Minisatellite mutation frequency in human sperm following radiotherapy. AB - Screening pedigrees for inherited minisatellite length changes provides an efficient means of monitoring repeat DNA instability but has given rise to apparently contradictory results regarding the effects of radiation on the human germline. To explore this further in individuals with known radiation doses and to potentially gain information on the timing of mutation induction, we have used an extremely sensitive single molecule approach to quantify the frequencies of mutation at the hypervariable minisatellites B6.7 and CEB1 in the sperm of three seminoma patients following hemipelvic radiotherapy. Scattered radiation doses to the testicles were monitored and pre-treatment sperm DNA was compared with sperm derived from irradiated pre-meiotic, meiotic and post-meiotic cells. We show no evidence for mutation induction in any of the patients and discuss this finding in the context of previous population studies using minisatellites as reporter systems, one of which provided evidence for radiation-induced germline mutation. PMID- 11006414 TI - DNA breakage detection-FISH (DBD-FISH) in human spermatozoa: technical variants evidence different structural features. AB - Non-irradiated and X-irradiated (80 Gy) human spermatozoa were processed for in situ DNA breakage detection-FISH (DBD-FISH) of the whole genome, following two alternative variations of the basic technique. In the first, cells were initially incubated in the alkaline unwinding solution for transformation of DNA breaks into single-stranded DNA (ssDNA) to be hybridized, followed by the lysing solutions for protein removal. In the second, incubation in the lysing solutions was carried out before the denaturation step. The first approach yielded two subpopulations. While most sperm nuclei were faintly labeled and had chromocenters, a small subpopulation was strongly and homogeneously labeled, due to extensive DNA breakage. X-ray exposure increased the surface and mean fluorescence intensity. Otherwise, when the denaturation step was performed after protein extraction, all sperm nuclei yielded strong and dispersed FISH signals. Protein removal allows access of the unwinding solution to the DNA, which has abundant alkali-labile sites, and thus gives rise to large areas of ssDNA that are labeled by FISH. X-ray exposure increased the dispersion of FISH signals but decreased their mean fluorescence intensity. A linear dose-response was generated using the second experimental variant, being 30 Gy the lowest dose for detecting induction of damage by X-rays in mature sperm chromatin. These results indicate that DBD-FISH is not only useful for in situ detection of DNA breakage but also for revealing structural features of chromatin. PMID- 11006415 TI - DNA breakage detection-fish (DBD-FISH): effect of unwinding time. AB - DBD-FISH is a new procedure that allows detection and quantification of DNA breakage in situ within specific DNA target sites. Cells embedded in an agarose matrix on a slide are treated in an alkaline unwinding solution to transform DNA breaks into single-stranded DNA (ssDNA). After removal of proteins, DNA probes are hybridized and detected. DNA breaks increase the ssDNA and relax supercoiling of DNA loops, so more probe hybridizes, thereby increasing the surface area and fluorescence intensity of the FISH signal. The probe selects the chromatin area to be analysed. In order to restrict the extension of unwound ssDNA to a region closer to the origin of the DNA break, human leukocytes were processed for DBD FISH with a whole genome probe, after a 10 Gy dose of X-rays, for various unwinding times: 5, 2 min and 30s. Two cell populations were detected after 30s, but not with the 5 or 2 min unwinding times. One cell group had small to medium haloes corresponding to the relaxation of DNA supercoiling after DAPI staining, and strong DBD-FISH labelling of induced DNA breaks, whereas the other cell group showed big haloes of DNA loop unfolding and an absence of DBD-FISH labelling. The latter group was similar to cells processed by DBD-FISH without the unwinding step. Thus, they should correspond to cells unaffected by the alkaline unwinding solution, possibly because very brief unwinding times do not allow the diffusion of the alkali into the cells deep within the gel, thus biasing the results. Taking this into account, 2 min seems to be the minimum unwinding time required for an accurate detection of a signal by DBD-FISH. PMID- 11006416 TI - DNA damage in rat lymphocytes treated in vitro with iron cations and exposed to 7 mT magnetic fields (static or 50 Hz). AB - The present study was undertaken to verify a hypothesis that exposure of the cells to static or 50 Hz magnetic fields (MF) and simultaneous treatment with a known oxidant, ferrous chloride, may affect the oxidative deterioration of DNA molecules. The comet assay was chosen for the assessment of DNA damage. The experiments were performed on isolated rat lymphocytes incubated for 3h in Helmholtz coils at 7 mT static or 50 Hz MF. During MF exposure, part of the cell samples were incubated with 0.01 microM H(2)O(2) and another one with 10 microg/ml FeCl(2,) the rest serving as controls. Lymphocyte exposure to MF at 7 mT did not increase the number of cells with DNA damage in the comet assay. Incubation of lymphocytes with 10 microg/ml FeCl(2) did not produce a detectable damage of DNA either. However, when the FeCl(2)-incubated lymphocytes were simultaneously exposed to 7 mT MF, the number of damaged cells was significantly increased and reached about 20% for static MF and 15% for power frequency MF. In the control samples about 97% of the cells did not have any DNA damage. It is not possible at present to offer a reasonable explanation for the findings of this investigation - the high increase in the number of lymphocytes showing symptoms of DNA damage in the comet assay, following simultaneous exposure to the combination of two non-cytotoxic factors -10 microg/ml FeCl(2) and 7 mT MF. In view of the obtained results we can only hypothesise that under the influence of simultaneous exposure to FeCl(2) and static or 50 Hz MF, the number of reactive oxygen species generated by iron cations may increase substantially. Further studies will be necessary to confirm this hypothesis and define the biological significance of the observed effect. PMID- 11006418 TI - Corrigendum to "Congenital abnormalities and indicators of germinal mutations in the vicinity of an acrylonitrile producing factory" PMID- 11006417 TI - High plating density improves Big Blue system efficiency without loss of sensitivity. AB - To increase efficiency in the Big Blue system, the plating density was increased from 15000 to 30000 or 45000 plaque forming units (pfus) per plate by increasing the density of the E. coli lawn and decreasing individual plaque size. Small plaque size ensured minimal overlap of the plaques. Liver from one 3- and one 25 month-old mouse (low and high mutation frequencies, respectively) was analyzed and neither plating density nor plaque size affected mutant/mutation frequency and pattern. The color intensity of particular mutant plaques was not affected by plaque size or plating density. Optimal sensitivity is achieved by sequencing mutants to calculate the mutation frequency from the mutant frequency and to identify altered patterns of mutation. Detailed effort and cost accounting of the Big Blue system (including mouse handling, DNA extraction, plaque screening, plaque purification, and DNA sequencing) reveals that one-quarter of the total effort is devoted to plating and screening of plates. This effort is reduced two fold with high plating density. The total cost of the Big Blue system is reduced by 17%. The total cost of the High Plating Density Big Blue system is now only 12% more costly than a selectable assay and offers an extensively validated system with a large mutation database representing a decade of effort. PMID- 11006419 TI - Detection of novelty by perinatal rats. AB - This study investigated the development of fetal/neonatal rats' ability to distinguish between a novel and familiar taste. Here, we report that neonatal rats alter their orofacial movements (e.g., mouth movements and licks) upon tasting saccharin (SAC) if it was experienced previously. We also sought to determine the origins and duration of this response. Fetuses of embryonic ages E17, E18, or E19 received an oral injection of 10 microL 0.3% SAC while in utero. These animals were then reexposed to SAC on postnatal day 3, (P3) and observations of orofacial motor responses were recorded. Only neonates that first experienced SAC on E19 exhibited a SAC-induced stimulation of mouthing and licking on P3. These data suggested that a taste-recognition memory (TRM) is maintained for up to 5 days (i.e., E19 to P3). However, in this paradigm, the youngest fetuses also have the longest retention interval. Could these data also reflect the limitations of the E17 and E18 fetuses in retaining the TRM? In a second study, we shortened the taste exposure-reexposure interval to 2 days in an attempt to detect the TRM in younger fetuses. As expected, E19 rats exhibited a TRM when tested on E21. However, neither the E17 nor E18 fetuses showed SAC induced increases in mouthing and licking when tested 2 days after their initial exposure (E19 or E20). Finally, in order to determine whether a TRM could be detected in fetuses as well as neonates (see above), we conducted an additional study wherein E21 fetuses were tested before parturition. Like E21 neonates, E21 rat fetuses that had received SAC on E19 showed a differential response to SAC depending on whether it was novel or familiar. Thus, although E21 fetal orofacial movements were less frequent than those of the E21 neonate, the fetal-testing procedures were not sufficient to obscure the detection of a TRM. In summary, the data indicate that E19 rat fetuses can acquire a TRM and retain it for at least 2 5 days, whereas E17 and E18 fetuses cannot. PMID- 11006420 TI - Entrainment of the rat motor activity rhythm: effects of the light-dark cycle and physical exercise. AB - The circadian system is believed to be composed of a population of oscillators that couple together and generate a single rhythm. If this coupling is not strong enough, the circadian system can be dissociated into two or more groups of oscillators, and this is manifested in a dissociation of the overt rhythm into at least two circadian components. This study aims to examine the influence of factors, such as the difference in impact between T and tau, light intensity, and access to a running wheel, on the distribution of motor activity throughout the light-dark (LD) cycle and the dissociation of the rhythm. Rats were submitted to LD cycles of 23 h (T23) or 25 h. For each such cycle, half the rats were submitted to high light intensity and the other half to low light intensity. For each of these conditions, half the rats were kept in small cages, and the other half were in cages with a running wheel. Rats were maintained first under LD cycles and afterwards under constant darkness (DD). Motor activity was recorded throughout the whole experiment by means of activity meters with infrared beams. Results show that the distribution of motor activity throughout the cycle and the after effects observed in the rhythm under DD depended on light intensity and access to the wheel. Moreover, under T23, some rats showed two simultaneous circadian components whose manifestation also depended on the experimental conditions. The results indicate that the strength of circadian entrainment to LD cycles in the rat depends on three factors: the period length of the LD cycle, light intensity used during the light phase, and access to a running wheel. PMID- 11006421 TI - Evaluation of two rodent delayed-response memory tasks: a method with retractable levers versus a method with closing doors. AB - The purpose of this study was to evaluate and compare two similar rodent memory tasks developed in our laboratory that employ stimulus discrimination and delayed response (light and tone stimuli and variable length delays) and to determine their sensitivity to the muscarinic-acetylcholine receptor (mAChR) antagonist, scopolamine hydrobromide (SCOP HBr), and its quaternary (methylbromide) analog (SCOP MBr). Male Wistar rats were trained in either an open chamber that employed retracting levers (RLM) during the delays, or a method that utilized closing doors (CDM) that separated the rats from the levers during delays to reduce positional (nonmnemonic) strategies. When the rats were well trained, dose-effect studies (microg/kg doses, s.c., 30 min before test sessions) of SCOP HBr or MBr were performed. Baseline performance was characterized by delay-dependent decreases in accuracy in both methods except when the tone was the stimulus in the RLM. SCOP HBr impaired performance in both tasks at the higher doses tested, although the effects were more consistent in the CDM task and accuracy associated with each stimulus was affected similarly. Surprisingly, SCOP MBr also impaired performance of each task, especially when the tone was the stimulus, while accuracy associated with the light was not affected in the CDM task. Overall, the results indicated that the CDM was a somewhat more reliable task, appearing to reduce positional strategies with less variability in response to the mAChR antagonists, although some stimulus-modality specific effects were noted. It also appears important to consider the peripheral effects of mAChR antagonists (and potential central effects of quaternary mAChR antagonists) when interpreting results from behavioral studies, especially those involving conditional discrimination and delayed response. PMID- 11006422 TI - Microinfusion of nefazodone into the basolateral nucleus of the amygdala enhances defensive behavior induced by NMDA stimulation of the inferior colliculus. AB - The inferior colliculus is notably associated with defensive behavior. Electrical or pharmacological stimulation of the inferior colliculus induces aversive reactions such as running and jumping. Lesion of the basolateral nucleus of the amygdala decreases the threshold of aversive reactions induced by electrical stimulation of the inferior colliculus. The present work examined the influence of microinjections of nefazodone, a serotonin (5-HT(2)) antagonist, into the basolateral nucleus of amygdala on aversive reactions induced by N-methyl-D aspartate (NMDA) microinjected into the inferior colliculus. Rats implanted with cannulae in the inferior colliculus and in the basolateral nucleus of the amygdala were submitted to the open-field test where defensive behaviors were observed. Results indicated that microinjection of nefazodone into the basolateral nucleus of the amygdala increases aversive responses induced by NMDA injections into the inferior colliculus. This result suggests that the inferior colliculus and the basolateral nucleus of the amygdala have a functional relationship on the neural circuitry of defensive behavior. Moreover, 5-HT(2) receptors located at the basolateral nucleus of the amygdala seem to play an inhibitory role on defensive behaviors induced by inferior colliculus stimulation. PMID- 11006423 TI - Effect of photoperiod on body weight gain, and daily energy intake and energy expenditure in Japanese quail (Coturnix c. japonica). AB - Effect of photoperiod and food duration on body weight gain, energy intake, energy expenditure, and sexual development were investigated in two strains of Japanese quail (Coturnix c. japonica), bred for meat (broilers) or egg production (layers), from 7 to 71 days of age. In a first experiment chicks were subjected to 18L:6D, 15L:9D, 12L:12D, 9L:15D, or 6L:18D, with ad lib food during the light period. In a second experiment birds were exposed to a long photoperiod (18L:6D or 15L:9D) with ad lib food during part of the light period (first 6 or 9 h, respectively). Longer photoperiods were associated with larger weight gains. In 18L:6D broilers total body weight gain was 262 g compared to 213 g in 6L:18D broilers. In layers, corresponding values were 182 and 131 g. This effect of photoperiod on weight gain was primarily due to the effect of photoperiod on food availability. The photoperiod below which detrimental effects on weight gain occurred was 9L:15D for both strains. Chicks subjected to 9L:15D or 6L:18D exploited crop filling to enhance energy intake. They also decreased nocturnal metabolic rates to a greater extent compared to levels during the light phase than chicks subjected to light periods of 12 h or more. Sexual maturation was stimulated by photoperiod. At the age of 71 days, eight out of nine females subjected to 18L:6D were producing eggs, but none of the 6L:18D females. It is concluded that changes in feeding behavior and energy expenditure shown under short photoperiods are part of a strategy that allow chicks to gain weight continuously. PMID- 11006424 TI - Selective depletion of bacteria alters but does not eliminate odors of individuality in Rattus norvegicus. AB - To determine if odors of individuality are influenced by the removal of Gram negative or Gram-positive gut bacteria, Long-Evans rats were trained in an operant olfactometer to discriminate between the odors of two individual conspecifics and their operant responses to three different odors in randomly presented probe trials were analyzed. Significantly more responses were made to the probe odors from two known individuals than to the probe odors from known individuals with their Gram-negative bacteria eliminated (Experiment 1) or their Gram-positive bacteria eliminated (Experiment 2). Responses to the probe odors from known rats with bacterial selectively depleted did not differ significantly from responses to probe odors from unknown rats. These results support the hypothesis that the urinary odor of an individual rat is altered by the removal of specific gut bacteria. In Experiments 3 and 4, subjects made fewer errors in learning to discriminate between the odors of the familiar rats whose bacteria had been selectively depleted than between the odors of unknown rats. This "savings effect" indicates that some components of the individual urinary odors were retained after the removal of specific gut bacteria. Thus, the eliminated bacteria were not totally responsible for the odors of individuality. The outcome of Experiments 3 and 4 also indicates that conclusions regarding the recognition of odors by rats should not be made on the outcome of probe trial experiments alone. PMID- 11006425 TI - Restraint stress augments antibody production in cyclophosphamide-treated mice. AB - These studies evaluated the effects of a psychological stressor (restraint, RST) on antibody production in male BALB/cByJ mice. In Experiment 1, mice were immunized with keyhole limpet hemocyanin (KLH, 100 microg i.p.) 8 h prior to 15 h of RST or food and water deprivation (FWD). RST mice exhibited higher serum anti KLH IgM and IgG antibodies than FWD mice. In Experiment 2, mice were given either cyclophosphamide (CY, 15 mg/kg) or saline (SAL) prior to immunization with KLH and RST or FWD. ANOVA revealed serum anti-KLH IgG antibody titers in CY+RST animals to be significantly higher than in CY+FWD, SAL+FWD, and SAL+RST mice. Anti-KLH IgM titers of CY+RST mice were higher than those of other groups before and after a second immunization with KLH. In Experiment 3, we show that these changes in antibody production are not likely to be mediated via CY-induced alterations in the reactivity of the hypothalamo-pituitary-adrenal axis to RST. Together, these results indicate two potentially immunomodulatory parameters (RST and CY) can interact to alter a humoral immune response. In addition, these data support the hypothesis that humoral immune response of mice can be more reactive to stress when the mice are given a low dose of an immunomodulatory drug prior to stressor exposure. PMID- 11006426 TI - The impact of milk-derived unconditioned and conditioned negative feedback on the microstructure of ingestive behavior. AB - We describe at the microstructural level the impact of unconditioned and conditioned negative feedback on the licking behavior of the rat. Six groups of rats were trained to ingest one of six different dilutions of sweetened milk (0.5:1, 1:1, 2:1, 4;1, 8:1, and 16:1, water to milk ratios) under real feeding conditions until intake was stable. Then they were given five sham-feeding tests with the same solution. We compared the size of the clusters (SC) and their number (NC) during the 15-min period when intake rate was declining in the real feeding test with SC and NC during the corresponding 15-min period in the following sham-feeding test. Intake increased significantly over the five sham feeding tests with the two highest concentrations, indicating the extinction of conditioned negative feedback. With these two solutions, we compared the microstructure of licking behavior in the first with that in the fifth sham feeding test to determine if conditioned negative feedback affected SC or NC or both. The effect of both unconditioned and conditioned negative feedback on licking behavior was to decrease the number of clusters without affecting their size. We conclude that negative feedback derived from the accumulation of milk in the gastrointestinal tract decreases the probability of initiating a bout of licking during a pause. It has no effect on the ability to continue a bout of licking once it has begun. PMID- 11006427 TI - Attention to metabolic hunger and its effects on Helicobacter pylori infection. AB - A significant decrease in the bacterial count of small intestinal mucosa has been observed in children with recurrent diarrhea or abdominal pain in the time that has elapsed from the previous meal. Humans may be trained to recognize metabolic feelings of hunger that are associated with a steady and slightly lower glycemia than baseline, between 4.7 and 3.9 mmol/L (intervention). An eating habit associated with a decrease in preprandial glycemia prevented diarrhea relapses, and was expected to impair intestinal microflora growth, including Helicobacter pylori in the stomach. The development of Helicobacter pylori infection might be prevented during childhood, and recovery from infection may be expected with intervention. The improvement in attention to metabolic feelings consisted of acquiring a predictive ability of glycemia by distinction between unsolicited hunger feelings (metabolic hunger) and those associated with external cues. Matching intake to the inbetween energy needs served to predict the subsequent emergence of the metabolic hunger. The matching was further compensated for the early or late emergence of metabolic hunger at the subsequent meals. Fruit and vegetables were increased to avoid abrupt glycemia lowering. This intervention was trained in 5-month periods. Subjects (209, 44, and 58) completed their training during 4-year periods between 1982 and 1994, and were enrolled in a prospective, controlled, randomized, interventional, preventive, and cohort study. The "prevention" hypothesis was tested in a subgroup of 86 healthy infants who were recalled in the years 1996 to 1998. A "recovery" study of approximately a 1-year intervention was investigated in 47 healthy subjects between ages 5 and 25, who were positive for anti-H. pylori and had no need for an immediate antibiotic treatment at entry. The following behavioral factors were recorded in a 7-day home diary and calculated: the fraction of meals induced by metabolic hunger out of 21 main mealtimes; average preprandial glycemia (DAP glycemia); daily intakes, activity; and bedtime hours. The decrease in preprandial glycemia was the objective measure of compliance with the recognition of "metabolic" hunger. Anthropometric measures and blood tests were obtained for nutritional and functional verifications. Average preprandial glycemia was 8.5 and 8.6% lower in the intervention groups than the control groups in the "prevention" and "recovery" studies, respectively, at the end of follow-up (p<0.05 and <0.001, respectively). A 4.7% seroprevalence of H. pylori infection was observed in the intervention group, with 30.2% in the control group at a mean age of 10 years after approximately an 8-year follow-up in the "prevention" study (p<0.0005). The seroprevalence decreased to 9 of 24 (37.5%) under intervention as opposed to 20 of 23 controls (87%) in the recovery study (p<0.002). A significant positive correlation was found between DAP glycemia and the anti-H. pylori serum antibody concentration (r = 0.52; p = 0.0002). A decrease in the level of immune stimulation by H. pylori infection was observed due to the intervention, which may have a preventive and therapeutic role on the infection. PMID- 11006428 TI - The relationship between audiogenic seizure (AGS) susceptibility and forebrain tone-responsiveness in genetically AGS-prone Wistar rats. AB - The present study characterized the intensity-response functions of extracellular field responsiveness of different cortical/subcortical structures of the forebrain following the free-field presentation of tone stimuli, within a population of genetically audiogenic seizure (AGS)-prone KM-Wistar rats. The neural response properties of each case were compared to its propensity to exhibit AGSs during the continuous tone stimulation (15 kHz, 90 s at max.). The amplitudes or slope components of the evoked responses and their peak latencies showed significant positive (amplitude and slope) and negative (peak latency) Bolzmann's sigmoid relationships with the onset-latency of AGS. These relationships, with areal differences in the slopes of saturation functions, applied for the three different data sets recorded simultaneously from the stratum radiatum dendritic layer of the hippocampal CA1 area, primary auditory cortex layers II-IV, and frontal cortex surface. In addition, the similar type of functions between the evoked response variables and AGS onset latency held when all the areas were considered together. These data suggest that the neural responsiveness to acoustic stimulation of the primary sensory, multimodal and association cortices of the forebrain may altogether contribute to the seizure initiation by that modality in the genetically AGS-prone rats. It has been previously shown that there exist abundant and dispersed auditory projections from these forebrain areas to the brain stem and spinal cord, structures that are generally considered to be the key predisposing factors in the generation of AGS. Hence, the types of correlation found reflect the subject-specific stage of forebrain responsiveness, being either related or unrelated to genetic AGS specific changes, and possibly its triggering impact upon the lower brain AGS network. Accordingly, the mere comparison of forebrain response measures of these AGS-prone animals with those of the AGS-resistant ones could not reveal the result presented. PMID- 11006429 TI - In two species, females exhibit superior working memory and inferior reference memory on the water radial-arm maze. AB - Male and female mice and rats were tested on a water escape version of the radial arm maze designed to measure working and reference memory. In both species, females exhibited superior working memory during acquisition, and were better able to handle a higher memory load. However, male mice and rats exhibited better reference memory than females during the asymptotic portion of testing. Our data suggest that females may be better at working memory when both working and reference memory information must be learned simultaneously, and males better at reference memory when it has been differentiated from working memory. PMID- 11006430 TI - The effect of activity history and current activity on static and dynamic postural balance in older adults. AB - The purpose of this study was to investigate the effects of former athleticism and current activity status on static and dynamic postural balance in older adults. Fifty-six subjects participated in four study groups including former athletes, currently active (AA; n = 15; 69.1+/-4.4 years.; 77.8+/-9.8 kg), former athletes, currently inactive (AI; n = 12; 66.7 years.; 87.2+/-15.1 kg), controls currently active (CA; n = 14; 68.6 +/- 4.5 years.; 73.9+/-15 kg), and controls currently inactive (CI; n = 15; 72.8+/-4.8 years; 81.1+/-14.8). All subjects were tested for height, weight, flexibility, thigh circumference, and static (sharpened Romberg/unipedal stance), and dynamic (step length and width) balance tests. The sharpened Romberg (eyes open) test showed that AA (60.0+/-0 s) and CA (59.4+/- 0.5 s) balanced significantly longer than AI (41.5+/-7.2 s), and CI (41.8+/-6.1 s) (p<0.05). The unipedal (eyes open) test balance scores for AA, CA, AI, and CI were respectively 40.0+/-4.5, 55.1+/- 3.4, 33.0+/-7.1, and 27.5+/-6.1 s, with CA significantly better than CI (p<0.05). In dynamic balance AA and CA (746.1+/-28.0 and 724.6+/-24.3 mm) showed significantly longer step lengths (p<0.05) than CI (643.7+/-26.5 mm). The eyes closed test results for relative group comparisons were similar. Overall, two-way analysis of variance showed a significant activity main effect for all dependent variables measured (p<0.05). The results indicated that current activity status plays a key role on balance performance in older adults. Furthermore, former athletic activity history provides no protection for the age related onset of postural imbalance. PMID- 11006431 TI - Short-term behavioral and electrophysiological consequences of underwater trauma. AB - In a previous work we found that a 30-s underwater trauma, following 8 days of training for a spatial memory task in the water maze, resulted in poor performance in the spatial memory task at both 1 h and 3 weeks after the trauma. Here we found that compared with naive animals and animals that were trained for the spatial learning task but were not traumatized, the traumatized rats showed impaired performance in a spatial learning task in the water maze 20 min after the trauma and a reduced level of dentate gyrus long-term potentiation (LTP) 40 min after high-frequency stimulation to the perforant path. We also found a positive correlation between the behavioral performance and hippocampal plasticity. The reduced ability to induce LTP suggests that the trauma-related behavioral impairment is mediated by hippocampal-dependent processes. The underwater trauma may provide an important and potentially powerful model for understanding the mechanisms underlying the relationship between stress, cognition, and learning. PMID- 11006432 TI - Effects of food on cortisol and mood in vulnerable subjects under controllable and uncontrollable stress. AB - The aim of this study was to investigate whether in stress-prone subjects, carbohydrate-rich, protein-poor food (CR/PP) diminished depressive mood and a cortisol response under controllable as well as uncontrollable laboratory stress. Twenty-two subjects with high stress proneness (HS) and 23 subjects with low stress proneness (LS) participated in a controllable- and uncontrollable-stress experiment during either a CR/PP or protein-rich, carbohydrate-poor (PR/CP) diet. Both controllable and uncontrollable laboratory stress significantly increased pulse rate and skin conductance in HS and LS subjects, whereas uncontrollable stress increased feelings of depression, anger, tension, and fatigue and decreased feelings of vigor. Only in HS subjects, a cortisol response and feelings of depression became lower under the CR/PP diet condition, irrespective of the controllability of the laboratory stressor, suggesting an increased ability to cope with stress. Because the CR/PP diet compared with the PR/CP diet previously has been found to cause a 42% increase in plasma tryptophan/SigmaLNAA, seen as an indirect measure of increases in brain serotonin levels, the present results suggest that an enhanced serotonin function in HS subjects may be involved. PMID- 11006433 TI - Palatability and intake relationships in free-living humans. characterization and independence of influence in North Americans. AB - In order to investigate palatability influences on the ad lib eating behavior of free-living humans, 564 participants were paid to maintain food intake diaries for 7 days. They recorded their intake along with a global rating of the palatability of the entire meal on a seven-point scale. It was found that most meals that are self-selected are palatable and that only 9.3% are rated as unpalatable. Meals that were highest in palatability were 44% larger than meals that were low in palatability, but palatability only accounted for around 4% of the variance in meal sizes. Multiple regression demonstrated that palatability appears to act on intake independent of the levels of other influential factors. These results were very similar to those observed for the French and suggest that palatability operates similarly on intake regardless of culture. Palatability appears to be an influence on the amounts ingested by free-living humans in their natural environments but appears to be only one of many influential factors and accounts for only a small proportion of the variance in intake. PMID- 11006434 TI - Influence of the shape of the experimental room on spatial learning in rats. AB - Rats and other mammals can orient themselves in their habitat and locate various goals using different learning strategies. When a cartographic or local strategy is used, the animals construct and store in their long-term memories a complex representation of the extramaze space. However, exactly what constitutes the behaviorally key components of the extramaze space is still little known. The purpose of this study was to investigate whether the shape of the environment that surrounds the animal is an informative factor when a cartographic strategy is used. Two experiments were performed in which the rats were trained to find food in the goal arm of a four-arm, plus-shaped maze. Experiment 1 shows that lesions to the hippocampus impair the acquisition of this task, which suggests that in fact, a cartographic strategy is necessary to solve it. In Experiment 2, in which only one group of neurologically intact rats was used, we directly manipulated the shape of the space around the maze. The results showed that once the animals have learned the task, the hiding of all the extramaze single landmarks while keeping intact the general shape of the experimental room does not affect performance. Only when the shape of the surrounding space was modified did the performance deteriorate significantly. These results are discussed in relation to the different components of the extramaze environment and the contribution of each of them to the spatial learning. PMID- 11006435 TI - Effects of prenatal stress on defensive withdrawal behavior and corticotropin releasing factor systems in rat brain. AB - Exposure of pregnant rats to stress results in offspring that exhibit abnormally fearful behavior and have elevated neuroendocrine responses to novelty and aversive stimuli. This study examined the effects of prenatal stress on plasma corticosterone, adrenal weight, defensive withdrawal behavior, and the density of receptors for corticotropin releasing factor (CRF) in the amygdala. Pregnant Sprague-Dawley rats were stressed by daily handling and saline injection (s.c., 0.9%, 0.1 mL) during the last week of gestation. Male offspring were studied at adulthood (60-120 days of age). Adrenal hypertrophy and increased plasma corticosterone were observed in the prenatally stressed offspring. Defensive withdrawal, an ethological measure of the conflict between exploratory behavior and retreat, was quantified in naive offspring, and in offspring exposed to restraint stress (2 h). Restraint stress increased defensive withdrawal in both control and prenatally stressed offspring. Both naive and restraint-stressed prenatally stressed offspring exhibited increased defensive withdrawal compared to control offspring. There was a significant interaction between prenatal stress and restraint stress, suggesting increased vulnerability of prenatally stressed offspring. The effects of restraint in the defensive withdrawal test were reduced by intracerebroventricular administration of the CRF antagonists, alpha-helical CRF9-41 (20 microg every hour) or D-phe(12), Nle(21, 38), C(alpha)-MeLeu(37)] CRF((12-41)) (5 microg every hour) during the restraint period. The difference between control and prenatally stressed offspring was abolished by the CRF antagonists, suggesting that increased activation of CRF receptors may be a factor in the behavioral abnormalities of prenatally stressed rats. Measurement of CRF receptors in amygdala revealed a 2.5-fold increase in binding in prenatally stressed offspring. In light of previous work from this laboratory demonstrating increased content and release of CRF in amygdala from prenatally stressed offspring, the present study suggests that the increased fearfulness of prenatally stressed rats may be a consequence of increased activity of CRFergic systems in the amygdala. PMID- 11006436 TI - Role of IL-6 in cytokine-induced sickness behavior: a study with IL-6 deficient mice. AB - Interleukin-6 (IL-6) is synthesized and released in response to the cytokine inducer lipopolysaccharide (LPS) and IL-1, and acts as an endogenous pyrogen. Systemic administration of LPS and IL-1 to mice induces signs of sickness, including reduction of social exploration, immobility and body weight loss. To assess the role of IL-6 in the induction of sickness behavior, male IL-6 deficient mice (IL-6 -/-, Balb/cAn genetic background) were used and compared to IL-6 +/+ littermates. The depressing effects of intraperitoneal LPS (2.5 microg/mouse) and IL-1beta (1.0 microg/mouse) on behavior and change in body weight were more marked in IL-6 +/+ than in IL-6 -/- mice. The same difference was observed when mice were injected with LPS (5 ng/mouse) and IL-1beta (1 ng/mouse) into the lateral ventricle of the brain (i.c.v.). These results show that IL-6 released at the periphery and /or in the central nervous system plays a role in the behavioral response to LPS and IL-1. PMID- 11006437 TI - Diurnal rhythms of blood pressure, heart rate, and locomotor activity in adult and old male Wistar rats. AB - The diurnal rhythms of systolic blood pressure (SBP), diastolic BP (DBP), heart rate (HR), and spontaneous locomotor activity (SLA) were determined in adult (6 month-old) and old (24-month-old) male Wistar rats by using radiotelemetry. The rhythm parameters (mesor, amplitude, acrophase, and percent rhythm) were analyzed by Fast-Fourier Transform and Cosinor methods. We found that the 12-h mean values of SBP, DBP, HR, and SLA were significantly (p<0.001) higher in the dark phase than in the light phase in both adult and old rats. The nocturnal 12-h mean value of HR was significantly (p<0.01) lower in old than in adult rats. In addition, the differences between diurnal and nocturnal 12-h mean values of SBP and HR were reduced in old rats. There was no significant difference in the 12-h mean values of SBP, DBP, and SLA between old and adult rats. Otherwise, the diurnal HR rhythm amplitude was significantly (p<0.01) reduced in old rats (32+/-2 vs. 46+/-2 bpm). A nearly 1-h delay in SBP and DBP acrophases (03h55 +/- 00h19 vs. 02h57 +/- 00h14 and 03h17 +/- 00h13 vs. 01h53 +/- 00h22, respectively) was found in old rats comparing to adult rats. No significant change in SLA diurnal rhythm was observed in old rats. In conclusion, these results show a decrease in the nocturnal 12-h mean value of HR and an alteration in cardiovascular diurnal rhythms by a 1-h delay of SBP and DBP acrophases and a reduction of HR rhythm amplitude in old Wistar rats. PMID- 11006438 TI - Parabrachial lesions disrupt responses of rats to amino acid devoid diets, to protein-free diets, but not to high-protein diets. AB - Normal rats "reduce" intake of diets that lack an essential amino acid (THR-DEV), are protein free (PO%), or contain a high proportion of protein (P75%). We tested the importance of the parabrachial nuclei (PBN) in signaling such adjustments of food intake by placing electrophysiologically guided lesions in these nuclei at points that responded to gustatory stimuli. When fed the THR-DEV diet, rats with PBN lesions (PBNx) decreased their food intake significantly less than the controls (78.5 vs. 44.4%). When put on a P0% diet, PBNx animals decreased their intake only 8% compared with 23% for our CONT group. When put on a P75% diet, however, both groups decreased their intake in an equivalent amount. These experiments show that the PBN is involved in the learned aversion to an amino acid devoid diet. PMID- 11006439 TI - The way in which the data are combined affects the interpretation of short-term feeding behavior. AB - Short-term feeding behavior of pigs has been analyzed using random process models and log-normal models. Both were successful despite very different underlying assumptions relating to the theory of control. Feeder visits of growing pigs, housed individually from 17 to 52 kg live weight, were recorded electronically over a continuous period of 35 days. For the combined data, intervals between visits to the feeder greater than 30 min could be described well by the negative exponential model. The starting probability of a visit was constant at around 0.3, suggesting randomness. Disaggregating the data for individual pigs or for individual weeks did not change this conclusion. Intervals in the day were of a different nature to those at night, and disaggregation of the data into these two periods revealed that the negative exponential model was not satisfactory for either period. The starting probability for both periods increased with time since the last visit. This is consistent with the idea of satiety. Therefore, the apparent randomness in the data pooled across the day and night is an artefact caused by pooling itself, and is not in conflict with the satiety concept. The implications of data handling are discussed with reference to studies of the physiological control of food intake. PMID- 11006440 TI - Spontaneous meal patterns in female rats with and without access to running wheels. AB - Rats display strong behavioral rhythms during the ovarian cycle. During estrus, food intake is minimal due to a decrease in meal size, and locomotor activity is maximal. To investigate how activity influences feeding patterns across the ovarian cycle, we used a computerized system to monitor spontaneous meal patterns in intact, cycling female rats with and without access to running wheels. We found that running wheel access decreased dark meal frequency, increased dark meal size, and increased 24-h water intake during each phase of the ovarian cycle. In contrast, body weight, 24-h food intake, and the ovarian rhythms of reduced food intake, meal size, and body weight during estrus were not affected by running wheel access. In particular, the reduction in food intake during estrus was due to a selective reduction in dark meal size, not dark meal frequency, and this occurred independent of wheel access. These data indicate that estrus-related changes in spontaneous meal patterns and locomotor activity are independently controlled and that the reduction in food intake during estrus involves a selective change in the neurobiological controls of meal size. PMID- 11006441 TI - Failure of neonatal clomipramine treatment to alter forced swim immobility: chronic treadmill or activity-wheel running and imipramine. AB - We examined whether chronic running on a treadmill or activity wheel would attenuate the increased swim immobility that has been reported after neonatal clomipramine (CLI) treatment. Male Sprague-Dawley pups (N = 60) were injected with the monoamine reuptake inhibitor clomipramine hydrochloride (40 mg/kg per day i.p.) from 8 to 21 days of age. Another group (N = 12) received saline vehicle. At age 4 weeks, the CLI pups were randomly assigned to experimental conditions: (1) sedentary; (2) 24-h access to an activity wheel; (3) sedentary that received the antidepressant drug imipramine hydrochloride (10 mg/kg twice daily) during the last 10 days of the experiment; (4) activity wheel + imipramine; (5) treadmill running (30 m/min for 1 h at 0 degrees incline, 6 days/week). At age 16 weeks, rats underwent the Porsolt swim test 48 h after the last imipramine injection and/or the last exercise session. The increase in swim immobility among CLI-treated rats was small (one quarter of SD) and not statistically significant (p>0.10). The results are not consistent with our previous finding of antidepressant-like effects of activity-wheel running based on brain noradrenergic adaptations and enhanced male copulatory performance after neonatal CLI treatment. The lack of change in swim performance after clomipramine questions the generalizability of the CLI model of depression and the validity of the forced swim test as a behavioral measure of depression when it is used after neonatal CLI injection or chronic activity-wheel running. PMID- 11006442 TI - Release of hypothalamic norepinephrine during MSG intake in rats fed normal and nonprotein diet. AB - Effects of monosodium L-glutamate (MSG) solution (0.06 M) on interstitial levels of norepinephrine (NE) were measured in the lateral hypothalamus (LH). Wistar male rats, housed in standard operant boxes, were fed either normal or nonprotein diet for 3 days. Beside a daily bar-mediated drinking session (75 min), animals were without access to fluids. Microdialysates, collected from the LH during the 75 min of drinking, were analyzed using high-pressure liquid chromatography. No significant responses of the LH NE to the drinking of distilled water, MSG, NaCl (0.06 M), and glucose solution (0.6 M) were found in normally fed rats. However, a specific decline in LH NE release was detected during MSG solution-drinking in rats fed nonprotein diet. As MSG preference indicates protein intake, it is possible that LH NE is, at least partially, one of the brain signals that relate MSG preference to dietary protein intake. PMID- 11006443 TI - Different pathways of neurohormonal hypothalamic control of the adrenal cortex function in young and old rats. AB - The pathways of hypothalamic control of the adrenal cortex function were studied in the experiments on young and old male Wistar rats. Immunohistochemical, morphometric, densitometric and radioimmunoassay methods were used. It was shown that in young rats under stress vasopressin (VP) is released into the portal circulation and probably in this way stimulates ACTH and enhances secretion of corticosterone in the adrenal cortex. In old rats stress-reaction is delayed and less adequate compared to young animals. It seems likely that in old rats VP is secreted into the general circulation mainly via the posterior pituitary lobe (PP), exerting a direct influence on the adrenal cortex. The conclusion is made that in old rats regulation of the adrenal cortex function is realised via the pathway phylogenetically more ancient than in young animals. PMID- 11006444 TI - Changes with aging in the modulation of macrophages by norepinephrine. AB - The effect of aging on the norepinephrine (NE)-induced modulation of phagocytic and oxygen-dependent microbicidal processes of mouse peritoneal macrophages was studied. Phagocytosis of latex beads on culture plates and superoxide anion production was evaluated in young (12 weeks), adult (22 weeks), mature (48 weeks) and old (72 weeks) BALB/c mice after in vitro incubation with 10(-12), 10(-9), 10(-7), 10(-5) or 10(-3) M concentrations of NE. The results indicate that the phagocytic response to NE is quite similar in young and mature mice, with increased phagocytosis after incubation with 10(-7) and 10(-3) M, and decreased phagocytosis with 10(-5) M. Macrophages from adult mice increased their phagocytic capacity after incubation with the highest concentrations of NE (10( 5) and 10(-3) M) and macrophages from old animals only were stimulated with 10( 3) M. In addition, it was found that usually NE increased the extracellular superoxide anion production in the absence of phagocytosis in adult mice. No statistically significant changes were found in intracellular superoxide anion levels, but an increase was seen after phagocytosis in macrophages from adult, mature and old animals, especially after incubation with 10(-5) M. In conclusion, the results obtained indicate that the modulation of macrophages by NE does not only depend on the concentration of this neurotransmitter, but also on age. PMID- 11006445 TI - Sponge homologue to human and yeast gene encoding the longevity assurance polypeptide: differential expression in telomerase-positive and telomerase negative cells of Suberites domuncula. AB - Porifera show a characteristic Bauplan in spite of the fact that (almost) all cells are telomerase-positive and presumably provided with an unlimited potency for cell proliferation. Studies revealed that telomerase-positive cells can be triggered to telomerase-negative cells by dissociating them into single cells. Single cells from the demosponge Suberites domuncula, in contrast to cells present in primmorphs (a special form of cell aggregates), lack the property to proliferate and they undergo apoptosis. One gene, SDLAGL, was identified in primmorphs that showed high sequence similarity to the longevity assurance genes from other Metazoa. In single cells no transcripts of SDLAGL could be identified, while high expression was seen after re-aggregation of single cells and in proliferating cells of primmorphs. We concluded that SDLAGL is involved in the shift of telomerase-positive, proliferating cells to telomerase-negative, non proliferating cells. PMID- 11006446 TI - Age-related changes in interferon-alpha/beta receptor expression, binding, and induction of apoptosis in natural killer cells from C57BL/6 mice. AB - Natural killer (NK) cells are a critical first line of defense against viral infections and tumors. We showed previously that basal NK cytotoxicity was comparable in adult (6 month) and aged (24 month) C57BL/6 (B6) mice. However, NK activity was significantly higher in adult compared with aged B6 mice after either in vitro or in vivo stimulation with IFN-alpha/beta. The present study explored whether age-related decreases in inducible NK activity after stimulation with IFN-alpha/beta were due to differences in (1) IFN-alpha/beta receptor expression or IFN-alpha/beta binding to NK cells or (2) apoptosis of NK cells. Flow cytometry revealed that, despite significantly higher IFN-alpha/beta receptor expression (P60 dB HL. Among the deaf-blind pupils and deaf infant pupils, there were several cases with rare hereditary syndromes. The prevalence of acquired causes of deafness, especially congenital rubella, had decreased over the years, whereas perinatal causes of deafness had increased. Chromosomal anomalies were found in 15% of the infant pupils in 1998. Over the study period, the percentage of pupils with multiple handicaps increased from 25 to 38%. PMID- 11006454 TI - Bilateral ethmoidal mucocele in cystic fibrosis: report of a case. AB - The purpose of this study is to report a rare and interesting case of a 10-month old boy who presented a bilateral ethmoidal mucocele associated with cystic fibrosis and to discuss, according to the literature, the sinonasal involvement in this disease. Only nine pediatric patients with both disorders have been reported previously in literature, and all of these cases were older than 1 year 4 months and presented with a unilateral mucocele. PMID- 11006455 TI - Nasopharyngeal carcinoma in children. AB - Nasopharyngeal carcinoma (NPC) is rare in children. We report two cases from KK Women and Children's Hospital in Singapore, who presented with advanced locoregional disease. The various aspects of NPC in children are discussed. It is more common in males and black adolescents. The etiology is probably different from that in adult. Undifferentiated NPC or lymphoepithelioma is the commonest variety. Confusing symptoms like cervical lymphadenopathy and unilateral otitis media with effusion (OME) are among the commonest presentations. When children present with both conditions simultaneously, it is imperative to manage them like adult patients and perform nasendoscopy to rule out NPC. The disease stage at presentation is often more advanced compared with adults. These tumors are, however, associated with a higher cure rate as they are most radiosensitive. Radiotherapy with neo-adjuvant chemotherapy is currently the treatment of choice. Both cases achieved satisfactory control of the disease and are presently under close follow-up. PMID- 11006456 TI - Prefabrication provides synapses on demand. PMID- 11006457 TI - Does the head rule the heart? PMID- 11006458 TI - Genotype x environment interaction: an old problem in a new field. PMID- 11006459 TI - Adult versus embryonic stem cells: which is the way forward? PMID- 11006460 TI - Behavioral neuroscience: challenges for the era of molecular biology. AB - This is a great age to participate in biological inquiry. Behavioral neuroscience offers a rich perspective for molecular biologists. However, behavioral analysis is not simply an assay. Whereas molecular biology has become a unique tool in the armamentarium of behavioral neuroscience, the powerful methodology of molecular biology is no substitute for careful behavioral exploration. PMID- 11006461 TI - Cell death in early neural development: beyond the neurotrophic theory. AB - The important effect of cell death on projecting neurons during development is well established. However, this mainstream research might have diverted recognition of the cell death that occurs at earlier stages of neural development, affecting proliferating neural precursor cells and young neuroblasts. In this article, we briefly present observations supporting the occurrence of programmed cell death during early neural development in a regulated fashion that to some extent parallels the death of projecting neurons lacking neurotrophic support. These findings raise new questions, in particular the magnitude and the role of this early neural cell death. PMID- 11006462 TI - Control of dorsal raphe 5-HT function by multiple 5-HT(1) autoreceptors: parallel purposes or pointless plurality? AB - The serotonergic cells of the dorsal raphe nucleus innervate much of the forebrain and are thought to be involved in the mechanism of action of antidepressants. Dysfunction of these cells might be involved in the neural mechanisms underlying depression and suicide. The traffic in pathways emanating from the dorsal raphe nucleus is controlled by 5-HT(1) autoreceptors. Until recently it was thought that the autoreceptors in the dorsal raphe nucleus were solely of the 5-HT(1A) subtype. In this article, we discuss evidence that the situation is more complex and that multiple 5-HT(1) subtypes govern different aspects of 5-HT function in the dorsal raphe nucleus presenting new therapeutic opportunities. PMID- 11006463 TI - G-protein-independent signaling by G-protein-coupled receptors. AB - Two classes of receptors transduce neurotransmitter signals: ionotropic receptors and heptahelical metabotropic receptors. Whereas the ionotropic receptors are structurally associated with a membrane channel, a mediating mechanism is necessary to functionally link metabotropic receptors with their respective effectors. According to the accepted paradigm, the first step in the metabotropic transduction process requires the activation of heterotrimeric G-proteins. An increasing number of observations, however, point to a novel mechanism through which neurotransmitters can initiate biochemical signals and modulate neuronal excitability. According to this mechanism metabotropic receptors induce responses by activating transduction systems that do not involve G-proteins. PMID- 11006464 TI - Common regions of the human frontal lobe recruited by diverse cognitive demands. AB - Though many neuroscientific methods have been brought to bear in the search for functional specializations within prefrontal cortex, little consensus has emerged. To assess the contribution of functional neuroimaging, this article reviews patterns of frontal-lobe activation associated with a broad range of different cognitive demands, including aspects of perception, response selection, executive control, working memory, episodic memory and problem solving. The results show a striking regularity: for many demands, there is a similar recruitment of mid-dorsolateral, mid-ventrolateral and dorsal anterior cingulate cortex. Much of the remainder of frontal cortex, including most of the medial and orbital surfaces, is largely insensitive to these demands. Undoubtedly, these results provide strong evidence for regional specialization of function within prefrontal cortex. This specialization, however, takes an unexpected form: a specific frontal-lobe network that is consistently recruited for solution of diverse cognitive problems. PMID- 11006465 TI - Live or let die - retinal ganglion cell death and survival during development and in the lesioned adult CNS. AB - Programmed cell death or apoptosis is a common and widespread phenomenon that is important for proper development of the nervous system. In the adult CNS, however, apoptosis contributes to secondary cell loss after various types of lesions. The retino-tectal system has been successfully used as a convenient model system to study the molecular mechanisms of neuronal apoptosis and survival during development and in the lesioned adult CNS. This review describes the current knowledge about the interactions of cell death and survival pathways in general and for retinal ganglion cells specifically. PMID- 11006467 TI - Context, state and the receptive fields of striatal cortex cells. AB - Visual cortical cells are commonly characterized by their receptive-field structure. Originally, a visual receptive field was defined in a purely spatial way as that retinal area from which a change in spiking response of the regarded cell could be elicited by visual stimulation. The first attempts to understand receptive-field structure were based entirely on the anatomical connectivity of the primary visual pathway. More recently, however, it has been discovered that the spatial and temporal context in which a stimulus is presented to a cell can strongly influence its receptive field, and this in turn is dependent on the state of arousal and attention. Accordingly, new concepts recognize that cortical receptive fields are highly dynamic entities embracing more than the sum of the full spatial and temporal response properties of a cell. PMID- 11006466 TI - New perspectives on enigmatic vanilloid receptors. AB - In spite of the rapid advances in our understanding of vanilloid-receptor pharmacology in the PNS, the function of vanilloid receptors in the brain has remained elusive. Recently, the endocannabinoid anandamide has been proposed to function as an endogenous agonist at the vanilloid receptor VR1. This is an exciting hypothesis because the localization of VR1 overlaps with that of anandamide and its preferred cannabinoid receptor CB(1) in various brain areas. The interaction of anandamide and/or related lipid metabolites with these two completely separate receptor systems in the brain clearly places VR1 in a much broader role than pain perception. At a practical level, the overlapping ligand recognition properties of VR1 and CB(1) might be exploited by medicinal chemistry. For example, arvanil, a 'chimeric' ligand that combines structural features of capsaicin and anandamide, promises to be an interesting lead for new drugs that interact at both vanilloid and cannabinoid receptors. PMID- 11006469 TI - The plasmodium vivax and P. berghei gene sequence tag projects PMID- 11006468 TI - Molecular approaches to malaria: seeking the whole picture. AB - This year, Australia hosted its first major international conference on malaria - Molecular Approaches to Malaria in Lorne, Victoria, 2-5 February 2000 (MAM2000). The worldwide research effort toward a better understanding of the pathogenesis and control of malaria in the post-genomic era was discussed and debated by over 250 researchers from 18 countries during four days packed with molecular biology, cell biology, genomics, vaccines and pathogenic mechanisms. This special malaria edition of Parasitology Today is an attempt to capture and summarize the quality and breadth of work presented at the conference and place this in the context of the current global malaria research effort; eight of the nine Reviews in this issue have been written by session chairs or presenters at MAM2000. PMID- 11006470 TI - Rupture and drug combinations on the Web PMID- 11006471 TI - Host cell invasion by malaria parasites. AB - The complex life cycle of the malaria parasite includes three specialized invasive stages, distinct both in terms of their cellular architecture and in their choice of target host cell. Despite the dissimilarities between these forms, there are clear parallels in the manner by which they enter their respective host cells. Advances in the area of erythrocyte invasion by the malaria merozoite, outlined here by Chetan Chitnis and Mike Blackman and discussed at the Molecular Approaches to Malaria conference, Lorne, Australia, 2 5 February 2000, will undoubtedly impact on our understanding of mechanisms of cell entry by the other invasive forms. Similarly, recent progress in dissecting the functional role of surface proteins expressed by sporozoite and ookinete stages has provided fascinating insights into general aspects of invasion by all invasive stages of apicomplexan parasites. PMID- 11006472 TI - Falciparum malaria: sticking up, standing out and out-standing. AB - Cytoadherence is believed to be fundamental for the survival of Plasmodium falciparum in vivo and, uniquely, is a major determinant of the virulence of this parasite. Despite the widely professed importance of cytoadhesion in the development of severe disease, there are a number of aspects of this highly complex process that remain poorly understood. Recent progress in the understanding of cytoadhesive phenomena was discussed extensively at the Molecular Approaches to Malaria conference, Lorne, Australia, 2-5 February 2000. Here, Brian Cooke, Mats Wahlgren and Ross Coppel consider just how far we have progressed during the past 30 years and highlight what is still missing in our understanding of the mechanisms and clinical relevance of this apparently vital process. PMID- 11006473 TI - Traffic jams: protein transport in Plasmodium falciparum. AB - Protein targeting in malaria parasites is a complex process, involving several cellular compartments that distinguish these cells from more familiar systems, such as yeast or mammals. At least a dozen distinct protein destinations are known. The best studied of these is the vestigial chloroplast (the apicoplast), but new tools promise rapid progress in understanding how Plasmodium falciparum and related apicomplexan parasites traffic proteins to their invasion-related organelles, and how they modify the host by trafficking proteins into its cytoplasm and plasma membrane. Here, Giel van Dooren and colleagues discuss recent insights into protein targeting via the secretory pathway in this fascinating and important system. This topic emerged as a major theme at the Molecular Approaches to Malaria conference, Lorne, Australia, 2-5 February 2000. PMID- 11006474 TI - A brief illustrated guide to the ultrastructure of Plasmodium falciparum asexual blood stages. AB - Interpretation of the new information arising from the Plasmodium falciparum Genome Project requires a good working knowledge of the ultrastructure of the parasite; however many aspects of the morphology of this species remain obscure. Lawrence Bannister, John Hopkins and colleagues here give an illustrated overview of the three-dimensional (3-D) organization of the merozoite, ring, trophozoite and schizont stages of the parasite, based on available data that include 3-D reconstruc-tion from serial electron microscope sections. The review describes the chief organelles present in these stages, emphasizing the continuity of structure in addition to specialized, stage-specific features developed during the asexual erythrocytic cycle. PMID- 11006475 TI - Malaria research in the post-genomic era. AB - Within the next few years, the complete genomic sequences of Plasmodium falciparum, and potentially several other Plasmodium spp, will be available to researchers worldwide. These complete genomic sequence data are certain to provide the foundation for nearly all malaria research in the next decades, as discussed here by Dan Carucci. PMID- 11006476 TI - Antimalarial drug development and new targets. AB - The Molecular Approaches to Malaria (MAM2000) conference, Lorne, Australia, 2-5 February 2000, brought together world-class malaria research scientists. The development of new tools and technologies - transfection, DNA microarrays and proteomic analysis - and the availability of DNA sequences generated by the Malaria Genome Project, along with more classic approaches, have facilitated the identification of novel drug targets, the development of new antimalarials and the generation of a deeper understanding of the molecular mechanism(s) of drug resistance in malaria. It is hoped that combinations of these technologies could lead to strategies that enable the development of effective, efficient and affordable new drugs to overcome drug-resistant malaria, as discussed at MAM2000 and outlined here by Ian Macreadie and colleagues. PMID- 11006477 TI - Malaria vaccines. AB - Although the possibility of a live attenuated malaria vaccine has been considered, current malaria vaccine development activities are dominated by attempts to develop a subunit vaccine. Hence, it is entirely appropriate that a session of the Molecular Approaches to Malaria conference, Lorne, Australia, 2-5 February 2000, was devoted to vaccine development. The oral presentations in this session and the relevant poster presentations are outlined here by Robin Anders and Allan Saul. PMID- 11006478 TI - Molecular approaches to epidemiology and clinical aspects of malaria. AB - Malaria is a problem of global importance, responsible for 1-2 million deaths per year, mainly in African children, as well as considerable morbidity manifested as severe anaemia and encephalopathy in young children. Fundamental to the development of new tools for malaria control in humans is an increased understanding of key features of malaria infection, such as the diversity of outcome in different individuals, the understanding of different manifestations of the disease and of the mechanisms of immunity that allow clinical protection in the face of ongoing low-grade infection (concomitant immunity or premunition). Here, Graham Brown and colleagues review some of the ways in which molecular approaches might be used to increase our understanding of the epidemiology and clinical manifestations of malaria, as discussed at the Molecular Approaches to Malaria conference (MAM2000), Lorne, Australia, 2-5 February 2000. PMID- 11006479 TI - Pathogenesis of malaria. AB - As the mortality rate of 20-30% for severe falciparum malaria under even the best clinical conditions testifies, access to antimalarial drugs is not sufficient to prevent an appreciable mortality from this disease. Understanding the cause of death at a cellular level is essential if additional rational treatments are to be developed. Here, Ian Clark and Louis Schofield discuss recent work presented at the Molecular Approaches to Malaria conference, Lorne, Australia, 2-5 February 2000, that updates the cytokine-based concept of malarial disease. PMID- 11006480 TI - Application of (quantitative) structure-activity relationships to progestagens: from serendipity to structure-based design. AB - Progestagens are drugs, which are widely used in hormonal contraception and in hormone-replacement therapy. Since the natural hormone, progesterone, lacks oral activity, much effort has been devoted to finding analogues with improved oral activity and, preferably, higher potency and selectivity. A crystal structure of the hormone binding domain (HBD) region of the progesterone receptor (PR) could only be obtained recently. For more than forty years the process of designing new progestagens could therefore only be guided by the knowledge of the structure of the ligand and its corresponding in vitro/in vivo activities. While in early days chemical intuition and simple statistics (structure-activity relationship - SAR) were leading the drug design process, in later days more complex statistics and visualization tools have become routinely part of quantitative structure-activity relationship (QSAR) studies. The present review aims to provide a general overview of the strategies, efforts and achievements of synthetic and computational chemists in more than forty years of development of progestagens. PMID- 11006481 TI - Synthesis of ionones and carvone analogues: olfactory properties and preliminary toxicity assays. AB - Vilsmeier reagents react with alpha/beta-ionones and carvone to produce aldehydes 7-11 in a one-step procedure. The indene derivative 11, which came from the double iminoalkylation of carvone and ring closure with the elimination of dimethylamine, was practically odourless, while all the others had peculiar odours which were very different from the starting material. The cytotoxicity data of 9 and 10, which are the most promising potential perfume ingredients, are also reported. PMID- 11006482 TI - Synthesis and cytotoxicity of gossypol related compounds. AB - Gossypol, gossypolone, reduced gossypol and new Schiff's bases of racemic gossypol and gossypolone were extracted or synthesized. Their cytotoxic activities on KB human cancer cells were determined. Gossypolone and the ethylamine derivative of gossypolone were the most active compounds (IC(50) in the micromolar range in both cases). The cytotoxicity of gossypol and gossypolone was increased when the tests were performed in the absence of serum and decreased when catalase as well as mannitol were added to the culture medium. PMID- 11006483 TI - Synthesis and beta-blocking activity of (R,S)-(E)-oximeethers of 2, 3-dihydro-1,8 naphthyridine and 2,3-dihydrothiopyrano[2, 3-b]pyridine:potential antihypertensive agents - part IX. AB - The synthesis of oximeethers of 2,3-dihydro-1,8-naphthyridine and 2, 3 dihydrothiopyrano[2,3-b]pyridine is described. These compounds exhibit a selective beta-blocking activity, with a selectivity towards beta(2)-receptors. Groups in the N(1) position giving rise to a considerable steric hindrance led to a higher beta(2)-blocking selectivity, whereas groups creating a moderate hindrance caused a weak but significant decrease in beta(2)-antagonist potency. Substitution of the N(1)-R group with a sulfur atom led to compounds possessing beta(1)-, beta(2)- and beta(3)-blocking properties. Compounds 9c(1) and 10a(1) showed a beta(3)-antagonist activity slightly lower than that of propranolol. PMID- 11006484 TI - Synthesis and antiproliferative activity of some variously substituted acridine and azacridine derivatives. AB - A group of 9-substituted acridine and azacridine derivatives (m-AMSA analogues) were synthesised following classical procedures as potential antitumour agents with inhibitory effects on DNA topoisomerase II. Some were found to have noticeable cytotoxicity against human HL-60 and HeLa cells grown in culture. Their non-covalent interactions with calf thymus DNA have been studied using fluorescence quenching. We evaluated DNA damage produced by the tested compounds by means of DNA filter elution and protein precipitation techniques. Catalytic studies carried out with purified topoisomerase confirmed these agents as antitopoisomerase inhibitors. Chemotherapy of solid-tumour-bearing mice with tested compounds allowed an aza-analogue (compound IIIb), as potent as m-AMSA but less toxic towards the host, to be recognised. PMID- 11006485 TI - Synthesis of (1S,3aS)-8-(2,3,3a,4,5, 6-hexahydro-1H-phenalen-1-yl)-1-phenyl-1,3,8 triaza-spiro[4. 5]decan-4-one, a potent and selective orphanin FQ (OFQ) receptor agonist with anxiolytic-like properties. AB - The development of 8-(2,3,3a,4,5, 6-hexahydro-1H-phenalen1-yl)-1-phenyl-1,3,8 triaza-spiro[4. 5]decan-4-ones 3 starting from (RS)-8-acenaphten-1-yl-1-phenyl 1,3, 8-triazaspiro[4.5]decan-4-one 1 is reported. The synthesis and the binding affinities at human OFQ and opioid (micro, kappa, delta) receptors of the stereoisomers 3a-f are described. In vitro the most selective compound, (1S,3aS) 8-(2,3,3a,4,5, 6-hexahydro-1H-phenalen1-yl)-1-phenyl-1,3,8-triaza-spiro[4. 5]decan-4-one 3c, was found to act as a full agonist at the OFQ receptor in the GTPgamma(35)S binding test. It turned out to be selective versus a variety of other neurotransmitter systems. When tested in vivo following intraperitoneal injection, compound 3c was found to decrease neophobia in a novel environment and to exhibit dose-dependent anxiolytic-like effects in the elevated plus-maze procedure, thus confirming the effects observed following intracerebroventricular infusion of the OFQ peptide in rat. PMID- 11006486 TI - Synthesis and antibacterial activity of some 5-guanylhydrazone/thiocyanato-6 arylimidazo[2,1-b]-1,3, 4-thiadiazole-2-sulfonamide derivatives. AB - 6-Arylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamides 3 on Vilsmeier-Haak reaction produced 5-formyl-6-arylimidazo[2,1-b]-1,3, 4-thiadiazole-2-[N (dimethylaminomethino)]sulfonamides 4, while 3 on treatment with potassium thiocyanate in the presence of bromine in acetic acid produced 5-thiocyanato-2 sulfonamides 6. Interaction of 4 with aminoguanidine hydrochloride in ethanol produced the corresponding 5-guanylhydrazone derivatives 5. Compounds 5 and 6 showed a high degree of antibacterial activity against both Escherichia coli and Staphylococcus aureus comparable to that of sulfamethoxazole and Norfloxacin. However, they were found to show moderate activity against Salmonella typhi, Pseudomonas aeruginosa and Pneumococci. PMID- 11006487 TI - Carbonic anhydrase inhibitors. Part 86. A QSAR study on some sulfonamide drugs which lower intra-ocular pressure, using the ACE non-linear statistical method. AB - Quantum chemical QSAR expressions have been developed for a heterogeneous group of 36 sulfonamides which have been shown to lower intraocular pressure in in vivo tests on animals. It was found, using the ACE statistical technique, that the lowering of intraocular pressure correlated non-linearly with K(I) for carbonic anhydrase inhibition and with solubility. Non-linear transformations had to be applied to both the response variable and to solubility. Chemical variables found to be relevant to CA inhibition included the dipole moment vector, the frontier orbital energies, the solvation energy determined by the COSMO model, the electrostatic potential based charges on the atoms of the sulfonamide group, and the size and polarizability of the molecule. PMID- 11006488 TI - Diagnostic imaging facilities for the general practitioner. PMID- 11006489 TI - Survey of the management of deep vein thrombosis in general practice in the Netherlands. AB - BACKGROUND: To investigate the management of (suspected) deep vein thrombosis in general practice. METHODS: Self completing postal questionnaire among a random sample of 692 general practitioners in the Netherlands. RESULTS: The overall response rate was 58%. Eighty-nine percent of the respondents initiated objective evaluation. Less than 3% usually make the diagnosis on clinical grounds only. Ninety-two percent initiated adequate treatment for the last patient with deep vein thrombosis. No more than 4% usually treat patients with acenocoumarol alone. Respondents frequently referred a patient to a specialist, 41% to confirm the diagnosis and 85% for treatment. Already 44% feel that management of deep vein thrombosis is a mandate of the general practitioner. For those who do not, the availability of diagnostic and therapeutic facilities are the main obstacles. CONCLUSION: In general practice objective diagnostic methods to evaluate suspected deep vein thrombosis are routinely used and patients receive adequate treatment. Although patients are frequently referred to the hospital many general practitioners feel that they should be able to take care of these patients themselves. (See Editorial p. 133) PMID- 11006490 TI - Availability of diagnostic facilities in the Netherlands for patients with suspected pulmonary embolism. ANTELOPE Study Group. Advances in New Technologies Evaluating the Localisation of Pulmonary Embolism. AB - Pulmonary embolism remains a complex diagnostic problem. Although accurate and cost-effective, the 'Dutch consensus' strategy is not widely applied. We assessed the availability and investment plans of the different facilities used in this strategy. Furthermore, the current and future availability of new diagnostic modalities was investigated. A questionnaire was sent to all Dutch hospitals. The questionnaire contained separate sections with questions for the hospital management and the medical practitioners at the departments of radiology, nuclear medicine, internal medicine and pulmonology. Five hundred and eighty-four questionnaires were sent out (response rate 68%). Forty-three percent of the hospitals had no nuclear medicine facility, 11% had no pulmonary angiography facility, and 59% had no spiral CT scan (SCTA). Forty-six percent of the responding hospitals had a nuclear medicine facility; and 5% used Technegas for ventilation studies. Strategies with SCTA were available in about 27% of the hospitals. Due to future investments this number will increase to approximately 55%. Strategies with Technegas were available in 2.4% of the hospitals, this number might increase to 25% if Technegas is proven accurate. The 'Dutch consensus' strategy is available in two-thirds of the hospitals. All other strategies were less feasible. Several equivalent strategies for diagnosing pulmonary embolism should be developed. These strategies should be accurate, widely available and accepted. PMID- 11006491 TI - Positron emission tomography for the detection of metastases of differentiated thyroid carcinoma. AB - OBJECTIVE: To show the value of positron emission tomography (PET) with 18-F fluorodeoxyglucose (18-FDG) for the detection of metastases of differentiated thyroid carcinoma in selected patients. PATIENT HISTORIES: There were four patients, who had undergone total thyroidectomy for papillary (two) or follicular thyroid carcinoma (two). All patients had subsequent treatment with (131)iodine. Three patients had an increasing serum concentration of thyroglobulin, one patient had antibodies against thyroglobulin. A diagnostic (131)iodine scintigraphy was negative in two patients, and uncertain in two patients. Positron emission tomography was performed about 45 min after administration of 10 mCi 18-F-fluorodeoxyglucose. In three patients PET showed uptake in the cervical region, caused by lymph node metastases in two (confirmed by neck dissection) and recurrent tumor on the trachea in one patient (confirmed by surgery). In the fourth patient uptake of 18-FDG was seen in the neck and in both lungs. This led to discontinuation of treatment with (131)iodine because the lung metastases did not accumulate (131)iodine. DISCUSSION: In selected patients with differentiated thyroid carcinoma with an increasing serum concentration of thyroglobulin, PET is an important diagnostic option when scintigraphy with (131)iodine is negative or uncertain. In the four presented case histories, the results of PET led to a therapeutic decision: surgery in three patients and discontinuation of (131)iodine in one patient. The development of guidelines for the use of PET in the diagnosis of recurrent thyroid cancer is discussed. PMID- 11006492 TI - Normal perfusion scintigraphy in pulmonary embolism. Causes and diagnostic alternatives. AB - Diagnosing pulmonary embolism (PE) is a challenge for many physicians as it is a frequently occurring disease with nonspecific symptoms and signs. Ventilation perfusion (V/Q) scintigraphy is widely used as the first step in diagnosing PE since it is non-invasive and highly sensitive. With a normal perfusion scan, clinically relevant pulmonary thrombo-emboli are considered to be absent. In an ongoing study assessing the value of spiral CT in the diagnosis of PE, we encountered a patient who had a normal perfusion scan while a large partially occluding thrombus in the right lower lobe artery and its branches was depicted by spiral CT and pulmonary angiography. In this article, we discuss the significance of normal findings in perfusion scintigraphy, the causes of false negative perfusion scans and the role of alternative techniques such as spiral CT and pulmonary angiography. PMID- 11006493 TI - An Epstein-Barr virus-associated pulmonary lymphoproliferative disorder as complication of immunosuppression. AB - Inherited or acquired immunodeficiencies as well as autoimmune diseases treated with cytotoxic drugs are associated with an increased incidence of lymphoma. Non Hodgkin's lymphomas that occur in the context of drug-induced immunosuppression, acquired or congenital immunodeficiency, are frequently associated with Epstein Barr virus infection. This report describes the occurrence of an Epstein-Barr virus associated pulmonary B cell lymphoma in a patient with longstanding rheumatoid arthritis treated with methotrexate. PMID- 11006494 TI - Disseminated intravascular coagulation as an unusual presentation of an Epstein Barr virus infection. AB - Epstein-Barr viral (EBV)-infection usually presents as fever, sore throat, fatigue, lymphadenopathy and atypical lymphocytosis. We describe a patient with disseminated intravascular coagulation as the presenting symptom caused by a primary EBV infection. PMID- 11006495 TI - The (non-)sense of the methionine loading test for detecting hyperhomocysteinaemia. PMID- 11006496 TI - Oestrogens and lower urinary tract function. AB - There is increasing evidence from animal and human studies that sex steroids have an important effect on the female lower urinary tract during adult life. Oestrogen receptors have been identified throughout the brain, pontine micturition centre and in the bladder, urethra and pelvic floor. Fluctuations in the circulating level of oestrogens and progesterone occurring during the menstrual cycle and in pregnancy influence the prevalence of urinary symptoms and the results of urodynamic investigation. In addition, the menopause and subsequent oestrogen deficiency have been implicated in the aetiology of a number of urogenital complaints including incontinence, urgency and recurrent urinary tract infection (UTI). However, the use of hormone replacement therapy for these conditions has given conflicting and largely disappointing results. The aim of this paper is to discuss the role of oestrogen in the pathogenesis and treatment of lower urinary tract dysfunction. PMID- 11006497 TI - Self-defined menopausal status in a multi-ethnic sample of midlife women. AB - OBJECTIVES: This study aimed to identify factors associated with women's perceived menopausal status and to evaluate agreement between women's self designation and a menstrually-based classification in a multi-ethnic sample of women. METHODS: A cross-sectional survey was conducted as part of a large, seven site, multi-ethnic study, the Study of Women's Health Across the Nation (SWAN). All variables were assessed by self-report in 13952 women aged 40-55 years. Multiple linear regression was used to assess determinants of self-defined menopausal status, stratifying by race/ethnicity within three anatomical/hormone use strata. Kappa statistics were used to evaluate agreement between the self defined and menstrually-based classifications. RESULTS: For women with an intact uterus, at least one ovary and not using hormones, menstrual patterns explained about half the variance in self-defined menopause status with older women classifying themselves later in the transition. Disagreement between menstrually based and self-defined menopausal status was 39, 38, 36, 32 and 29% for Hispanic, African-American, Japanese, Caucasian, and Chinese women, respectively (kappa statistics=0.46, 0.41, 0.40, 0.53 and 0.58). Women with vasomotor symptoms tended to self-designate themselves as being in transition regardless of their menstrual patterns. Age and 12 months of amenorrhea explained about 40% of the variance in self-categorization among women using hormones with an intact uterus. Bilateral oophorectomy, age and time since surgery explained about 20% of the variance among post-surgical women. CONCLUSIONS: Menstrual characteristics are strong predictors of women's self-perceived menopausal status. However, additional factors, including symptoms and cultural differences in the meaning of specific bleeding patterns, are also relevant and require further investigation. PMID- 11006498 TI - Relationships between catastrophic thoughts, perceived control and distress during menopausal hot flushes: exploring the correlates of a questionnaire measure. AB - OBJECTIVES: Many studies have established that highly negative or catastrophic thoughts about chronic health problems such as pain are associated with greater distress, lower self-efficacy for dealing with the problem and depressed mood. This study examined whether highly negative (or 'catastrophic') appraisals of hot flush experiences were associated with greater distress and lower perceived control regarding this menopausal problem. DESIGN: A postal survey was carried out, with a follow-up 12 months later. METHODS: Two questionnaires were initially completed by a volunteer sample of 61 women currently experiencing hot flushes. A mixture of qualitative and quantitative data were collected, including a Catastrophic Thoughts Questionnaire (CTQ) measure of catastrophic thoughts, based in part on Rosenstiel AK and Keefe FJ. The use of coping strategies in chronic low back pain patients: relationship to patient characteristics and current adjustment. Pain 1983;17:33-44. Thirty nine women who continued to report hot flushes were followed up 12 months later. RESULTS: Ratings to each item of the CTQ were highly intercorrelated. Women reporting more catastrophic thoughts tended to be lower in perceived control over flushes. Respondents' reported distress during hot flushes was more closely related to their frequency of negative thoughts about the problem, than to perceived control, flush chronicity or daily frequency of flushes. Over 12 months, respondents showed a highly stable pattern of catastrophic thoughts, and continued links with levels of distress during flush episodes. CONCLUSIONS: Cognitive appraisal processes that undermine coping with other chronic health problems seem also relevant to understanding the distress triggered by intermittent, unpredictable flush episodes. The findings imply that women may develop improved tolerance for menopausal flushing through challenging negative interpretations of the experience. PMID- 11006499 TI - Effects of estrogen therapy on well-being in postmenopausal women without vasomotor complaints. AB - OBJECTIVE: To establish whether estrogen treatment affects well-being in postmenopausal women without current or previous vasomotor symptoms. DESIGN: Forty postmenopausal women, aged 45-59 years, without current or previous vasomotor complaints, were included. They were randomized to masked treatment with either transdermal 17beta-estradiol 50 microg/24 h or to placebo. At baseline and after 12 and 14 weeks of treatment, the women completed a questionnaire which reflects well-being, the Psychological General Well-Being (PGWB) Index. RESULTS: The women scored high on the PGWB Index, both at baseline and after 12 and 14 weeks of treatment. There was no significant difference in well-being according to PGWB Index between the groups treated with estrogen and placebo, neither at baseline, nor after therapy. Furthermore, there was no difference in change during therapy between the treatment groups. CONCLUSION: There is a gradual decline in estrogen during the climacteric, and it is controversial to which extent this affects women's mental health. The PGWB scores in this study were high before therapy, reflecting that these women without previous or current vasomotor complaints represented a selected sample. Neither short-term estrogen treatment over 12 weeks nor addition with medroxyprogesterone acetate during 2 weeks improved well-being in postmenopausal women without vasomotor symptoms who had high well-being at baseline. PMID- 11006500 TI - Expression of estrogen receptor (ER) in oral mucosa and salivary glands. AB - OBJECTIVES: To assess the expression of estrogen receptor (ER) in oral mucosa and salivary glands, buccal mucosal biopsies from ten postmenopausal women (taken before and during the hormone replacement therapy), as well as, single biopsies from 20 healthy 19-year-old women were analyzed for ER expression. Salivary gland biopsies were taken from the minor labial salivary glands (n=6), submandibular glands (n=5) and parotid gland (n=1) from women at different ages. METHODS: total RNA extracted from the tissue samples was reverse-transcripted (RT) to single stranded cDNA, and the RT-polymerase chain reaction (RT-PCR) product was subjected to nucleotide sequencing to confirm the match with ER cDNA. Immunohistochemistry (IHC) with a monoclonal ER antibody (ER-ICA, Abbott) and Western blot analysis with monoclonal antibody against ER-related antigen (ER-D5, Amersham) were performed on the biopsies taken from the postmenopausal women. RESULTS: ER mRNA was expressed in 18/20 (90%) and 20/20 (100%) of the mucosal biopsies in the postmenopausal and 19-year-old women, respectively. The expression of mRNA was detected in all the submandibular gland samples, in the single parotid gland, as well as, in 4/6 (67%) of the labial glands. ER expression could not be detected by IHC, indicating either a very low level of expressed protein or difficulties in recognizing the epitopes by IHC. However, Western blot demonstrated 8/8 (100%) of the mucosal biopsies of postmenopausal women positive for ER-related antigen. CONCLUSIONS: the presence of ER mRNA and immunoreactive ER protein suggests that estrogens have a biological role in oral mucosa and salivary glands. PMID- 11006501 TI - Effects of norethisterone acetate addition to estradiol in long term HRT. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy and tolerability of hormone replacement therapy (HRT) among postmenopausal women living in the Lund area of Southern Sweden and to analyze treatment effects in different types or routes of HRT administration, as well as to compare with unopposed estrogen therapy. METHOD: in an ongoing, large population-based, prospective cohort study, this interim analysis included 3900 women. Of them, 693 postmenopausal women were eligible in the present analyses as they continued to use one of the four commercial HRT products for at least 2-3 years, i.e. continuous oral estradiol (E(2)) 2 mg+norethisterone acetate (NETA) 1 mg (CON-O), sequential oral estradiol 2 mg + norethisterone acetate 1 mg (CYC-O), sequential transdermal estradiol 50 microg + norethisterone acetate 250 microg (CYC-TRANS) and estradiol monotherapy. These women completed one generic questionnaire and one specific 'hormonal' questionnaire, as well as a personal interview pertaining to socio-demographics, detailed status of HRT use, and therapeutic efficacy and untoward side-effects by HRT. RESULTS: comparing the three combined E(2)+NETA groups with E(2) monotherapy, the beneficial effects on sexual desire and emotional well-being were significantly less in the combined groups than in E(2) monotherapy group. There was no significant difference regarding the negative side-effects between the groups. No significant difference was found between CON-O and CYC-O groups either in positive effects or in negative side-effects. A higher prevalence of positive effects was found in CYC-TRANS group than that in CYC-O group, especially in amelioration of sleep and urinary symptoms. Higher odd ratios of negative effects by HRT, such as irregular bleeds, weight gain, food craving and skin disorders were also found in CYC-TRANS group. CONCLUSION: in long-term HRT administration, the addition of a progestogen in HRT could compromise the beneficial effects of estradiol, particularly, the effects on women's emotional well being and psychosexual functioning. Administration of NETA continuously and sequentially had similar therapeutic efficacy and tolerability. More marked positive effects, such as improving of sleep and urinary symptoms, as well as nuisance side-effects, i.e. irregular bleeds, weight gain, food craving and skin disorders were encountered by the women using sequential transdermal regimen PMID- 11006502 TI - Collagen: image processing and quantitative ultrastructural studies PMID- 11006503 TI - Type V collagen: heterotypic type I/V collagen interactions in the regulation of fibril assembly. AB - Type V collagen is a quantitatively minor fibrillar collagen with a broad tissue distribution. The most common type V collagen isoform is alpha1(V)(2) alpha2(V) found in cornea. However, other isoforms exist, including an [alpha1(V)alpha2(V)alpha3(V)] form, an alpha1(V)(3) homotrimer and hybrid type V/XI forms. The functional role and fibrillar organization of these isoforms is not understood. In the cornea, type V collagen has a key role in the regulation of initial fibril assembly. Type I and type V collagen co-assemble into heterotypic fibrils. The entire triple-helical domain of the type V collagen molecules is buried within the fibril and type I collagen molecules are present along the fibril surface. The retained NH(2)-terminal domains of the type V collagen are exposed at the surface, extending outward through the gap zones. The molecular model of the NH(2)-terminal domain indicates that the short alpha helical region is a flexible hinge-like region allowing the peptide to project away from the major axis of the molecule; the short triple-helical regions serve as an extension through the hole zone, placing the tyrosine-rich domain at the surface. The assembly of early, immature fibril intermediates (segments) is regulated by the NH(2)-terminal domain of type V collagen. These NH(2)-terminal domains alter accretion of collagen molecules onto fibrils and therefore lateral growth. A critical density would favor the initiation of new fibrils rather than the continued growth of existing fibrils. Other type V collagen isoforms are likely to have an important role in non-cornea tissues. This role may be mediated by supramolecular aggregates different from those in the corneal stroma or by an alteration of the interactions mediated by tissue-specific type V collagen domains generated by different isoforms or aggregate structures. Presumably, the aggregate structure or specific domains are involved in the regionalization of fibril-associated macromolecules necessary for the tissue-specific regulation of later fibril growth and matrix assembly stages. PMID- 11006504 TI - Fourier and power law analysis of structural complexity in cornea and lens. AB - The ordered pattern of type I collagen fibrils in the transparent cornea is an example of specialization in the formation of functional ultrastructure. In contrast, the disordered and amorphous distribution of cytoplasmic proteins in the transparent lens resembles the structure of most cells. While the organization of cytoplasmic proteins is often considered to be random, the compartmentalization of functional proteins in biological cells and the organization provided by cytoskeletal elements suggests that non-random patterns of organization are common. Attempts to quantify disordered, amorphous patterns of ultrastructure in cells and tissues have been unsuccessful, in part, because the cellular organization of structural proteins including collagen, keratin, cytoskeletal and crystallin proteins is complex. Characterization of the complex patterns observed in electron micrographs is a fundamental problem in structural biology. This paper reviews the use of Fourier and power law analyses of electron micrographs of cornea and lens as models for ordered and disordered ultrastructure of cells and tissues. PMID- 11006505 TI - Collagen structure and functional implications. AB - The bio-mechanical requirements to which the connective tissue is subjected suggest that a causal correlation exist between the substructure and the collagen fibril function. We discuss the relationship between the inner structure of collagen fibrils, their diameter, their spatial layout and the functional requirements they have to withstand, and suggest that collagen fibrils may belong to two different forms indicated as "T-type" and "C-type". The first class, consisting of large, heterogeneous fibrils, parallely tightly packed, subjected to tensile stress along their axis is found in highly tensile structures such as tendons, ligaments and bone. The other class, consisting of small, homogeneous fibrils, helically arranged, resisting multidirectional stresses, is mostly present within highly compliant tissues such as blood vessel walls, skin and nerve sheaths. What causes these architectures to appear is discussed in detail in this review. PMID- 11006506 TI - Corneal and scleral collagens--a microscopist's perspective. AB - This paper reviews our existing understanding of the distribution and organisation of collagen types within the corneal and scleral stroma from a microscopical perspective. The contribution of various types of light microscopy, electron microscopy and atomic force microscopy to this field are separately discussed. Light microscopy was used in the earliest studies of the cornea and lead to the first description of the lamellar structure of the stroma. More recently polarised light microscopy has been used to obtain specific information on fibril orientation within individual lamellae. Light microscope immunolabelling techniques have been utilised to determine the distribution of several collagen types within the cornea and sclera, while recent developments in confocal microscopy have allowed detailed observations to be made on live cornea. Scanning electron microscopy has proved useful in determining the 3D organisation of lamellae within both corneal and scleral stroma. The transmission electron microscope was responsible for first revealing the regular diameter and high degree of order of the collagen fibrils present in the corneal stroma and contrasting this with the irregular diameter of fibrils present in sclera. This finding lead directly to the formulation of a theory of corneal transparency based on the uniformity of fibril diameter and packing. The use of specialised stains such as cuprolinic blue allowed direct observation of the glycosaminoglycan chains on proteoglycan molecules in cornea and sclera. These images allowed the binding sites of the proteoglycans along the collagen fibrils to be identified and provided convincing evidence for the importance of the proteoglycan molecules in collagen fibril organisation. Immunogold labelling has been used to map the distribution of several collagen types within the corneal and scleral stroma at the ultrastructural level and provided critical evidence for the role of type V collagen in the regulation of fibril diameter within the cornea. Specialised freezing-etching techniques have revealed the surface features of the collagen fibrils in corneal stroma, indicating clearly the presence of crossbridge structures between fibrils. The technique of rotary shadowing has been used to determine the conformation of several collagen types. In more recent years atomic force microscopy has been applied to the study of the corneal stroma. It has largely confirmed the observations made by the transmission electron microscope and provided independent evidence of crossbridge structures between the collagen fibres in cornea and sclera. The full potential of this technique has yet to be realised. PMID- 11006507 TI - STEM/TEM studies of collagen fibril assembly. AB - Quantitative scanning transmission electron microscopy (STEM), implemented on a conventional transmission electron microscope with STEM-attachment, has been a primary tool in our laboratory for the quantitative analysis of collagen fibril assembly in vivo and in vitro. Using this technique, a precise measurement of mass per unit length can be made at regular intervals along a fibril to generate an axial mass distribution (AMD). This in turn allows the number of collagen molecules to be calculated for every transverse section of the fibril along its entire length. All fibrils show a near-linear AMD in their tip regions. Only fibrils formed in tissue environments, however, show a characteristic abrupt change in mass slope along their tips. It appears that this tip growth characteristic is common to fibrils from evolutionarily diverse systems including vertebrate tendon and the mutable tissues of the echinoderms. Computer models of collagen fibril assembly have now been developed based on interpretation of the STEM data. Two alternative models have so far been generated for fibril growth by accretion; one is based on diffusion limited aggregation (DLA) and the other based on an interface-limited growth mechanism. Inter-fibrillar fusion can also contribute to the growth of fibrils in vertebrate tissues and STEM data indicates the presence of a tight regulation in this process. These models are fundamental for the hypotheses regarding how cells synthesise and spatially organise an extracellular matrix (ECM), rich in collagen fibrils. PMID- 11006508 TI - Structural variations of collagen in normal and pathological tissues: role of electron microscopy. AB - The spectrum of ultrastructural appearances assumed by collagen in normal and pathological tissues is illustrated using techniques of thin section transmission electron microscopy and computer-assisted analysis. The normal fibrillar collagen types are described in order to provide a basis for comparing other normal and abnormal forms. In normal tissues, the anchoring fibril and basal lamina (basement membrane) represent tissue structures largely containing collagen but differing significantly in organisation from normal types I to III fibrillar collagen. In pathological tissue, deviations from normal fine structure are reflected in abnormal aggregates of collagen fibrils (amianthoid and skeinoid fibres) and abnormalities in fibril diameter and cross-sectional profile. Fibrous and segment long-spacing collagen represent two further organisational variants of collagen, the former found widely in pathological tissues, the latter very rarely. Much remains to be discovered about these abnormal collagen variants their mode of formation, the cells that produce them, and their roles. They also present a challenge for the collagen biologist formulating hypotheses of collagen fibril assembly and molecular organisation. PMID- 11006509 TI - Collagen fibril organisation in mammalian vitreous by freeze etch/rotary shadowing electron microscopy. AB - Mammalian vitreous gel contains two major network-forming polymeric systems: long, thin fibrils comprising predominantly type II collagen and a meshwork of hyaluronan. The gel structure is maintained primarily by the collagen component, but little is known about the mechanisms of spacing of the collagen fibrils and of interactions between fibrils to form a stable network. In this study we have applied the technique of freeze etching/rotary shadowing electron microscopy in order to reveal the fibrillar network in central, cortical and basal vitreous and to understand the structural relationship between the collagen fibrils. The fibrils were arranged side by side in narrow bundles that frequently branched to link one bundle to another. Only a minor part of the fibrillar network consisted of segments that had a diameter of a single fibril (16.4nm mean diameter). In addition, three morphologically distinct filamentous structures were observed that appeared to form links within the collagen fibrillar network: short, single interlinking filaments of 7.0nm mean diameter, network-forming filaments of 6.7nm mean diameter, and longer filaments of 8.2nm mean diameter. All three types of filamentous structure were removed by digestion of the vitreous gels with Streptomyces hyaluronan lyase prior to freeze etching, indicating that these structures contain or are stabilised by hyaluronan. These filamentous structures may contribute to the structural stability of the vitreous gel. PMID- 11006510 TI - Collagen as a model system to investigate the use of aspirin as an inhibitor of protein glycation and crosslinking. AB - Aspirin has been shown to be a powerful inhibitor of post-Amadori Maillard reactions, although the exact mechanism of this action remains unclear. We have used corneal and scleral collagen as a model system: (i) to assess how aspirin, either alone or in combination with sugars, affects the surface charge distribution along the collagen fibrils; (ii) to see how sugars and/or aspirin affect the swelling properties of the cornea; and (iii) to see if sugars and/or aspirin change the distribution of water within the corneal stroma as the tissue swells. Charge changes were detected by examining changes in the uptake of phosphotungstate ions as seen in the electron microscope. Swelling was measured by monitoring the uptake of water as a function of swelling time, and water distribution was determined by using synchrotron X-ray diffraction to follow changes in the interfibrillar Bragg spacing as the cornea swells. Aspirin has a marked effect on the positive staining pattern of scleral collagen. This is different to the changes in stain uptake produced by glycation. Incubation with both sugars and aspirin showed that the sugar binding takes precedence over the effects of aspirin which, in turn, suggests that protein acetylation by aspirin is unlikely under these circumstances. However, aspirin completely suppresses corneal swelling. Even when the aspirin is removed, swelling in distilled water is reduced, and this is accompanied by changes in the water distribution. The results suggest that water is more evenly distributed in aspirin-treated corneas that are subsequently swollen than in swollen glycated corneas. Fructation, glucation and ribation on their own have little effect on the uptake of water as the cornea swells. This suggests that any sugar-derived crosslinks formed at this stage do not limit swelling. PMID- 11006511 TI - The fibril structure of type V collagen triple-helical domain. AB - Although the triple-helical structure of fibrillar collagen is regarded in general as being quite similar, each type of collagen molecule has inherent characteristics in the triple-helical domain. Few studies have ever been performed in terms of the aggregate structure of the triple-helical domain of fibrillar collagen. Reconstituted aggregates from the purified triple-helical domain of each type of fibrillar collagen might amplify the subtle differences in the structural characteristics of each type of collagen molecule. In this study, the reconstituted aggregate structure of pepsin-treated type V collagen (type Vp collagen), that is, virtually its triple-helical domain was characterized by transmission electron microscopy. Pepsin-treated type I (type Ip) and type II (type IIp) collagen were compared with type Vp collagen. Unique features of the aggregate structure of the triple-helical domain of the type V collagen can be summarized as follows:These results suggested that the lateral packing of the triple-helical domain of type V collagen is determined by its molecular structure. The characteristics of type Vp collagen fibrils might be explained by their characteristic amino acid composition. A significant feature of the triple helical domain of type V collagen is the high content of glycosylated hydroxylysine residues. Molecular model building of the collagenous structure suggests that a change in surface roughness is conspicuous by incorporating the glycosylated hydroxylysine residues. More than a ten-fold content of bulky glycosylated hydroxylysine residues in type V collagen compared to that of type I might have a significant influence on both the intermolecular and interfibrillar interactions of the triple-helical domain of type V collagen molecule. PMID- 11006512 TI - Thermally labile domains in the collagen molecule. AB - We have proposed that the denaturation kinetics of the characteristic sharp melting point of the collagen molecules is an irreversible rate rather than an equilibrium process as previously believed. This leads to the concept of domains of variable thermal stability along the length of the molecule. We have identified the major thermally labile domains from which the denaturation process is initiated as hydroxyproline deficient sequences of 65, 65 and 59 residues near the carboxy terminus in fibrillar collagen types I, II and III, respectively. These domains differ in that there is a single hydroxyproline in the type II domain and two hydroxyprolines in the type III domain. Similar sized domains are conserved in these collagen types across species including amphibians and invertebrates. The effective size of the domain is reduced in the fibrillar aggregates to 26 residues due to the interaction with adjacent molecules and because of the precise quarter-staggered alignment of the molecules the domains are located in the gap region. This spatial confinement within the lattice of the fibre leads to the significant increase in denaturation temperature of the fibre compared to the molecule. These labile domains have also been located in molecules that form the non-fibrillar type IV basement membrane collagens and the fibril-associated aggregates such as type IX. Based on the location of the different domains in type IX we have proposed a different arrangement of the type IX on the type II fibril. The model stresses the importance of hydroxyproline in stabilising the triple helix and supports the concept of hydrogen-bonded water bridges originally proposed from X-ray diffraction studies in contrast to other studies indicating water-bridges do not play a role in stabilising the collagen molecule. PMID- 11006513 TI - Diameter distributions of collagenous tissues in relation to sex. A quantitative ultrastructural study. AB - A number of factors have been implicated in the regulation of collagen fibril diameter. Electron microscopy analysis was used to investigate the role of sex on fibril diameter. Female mouse skin collagen fibrils' mean diameter values were significantly smaller than those from the male, independent of age. In addition, the female rabbit collagen fibrils had a marked decrease in the mean diameter compared to male, in all the tissues examined (skin, liver, and bone). These data suggest that the collagen fibril diameter is related to sex. PMID- 11006514 TI - Measurement of the axial periodicity of collagen fibrils using an image processing method. AB - In this paper, a method is described based on a computer-aided analysis of electron-optical images of collagen fibrils from various tissues, in order to determine the axial periodicity of such fibrils. The method gives information at a level of 2-3nm. PMID- 11006515 TI - A study of fibrous long spacing collagen ultrastructure and assembly by atomic force microscopy. AB - Fibrous long spacing collagen (FLS) fibrils are collagen fibrils that display a banding with periodicity greater than the 67nm periodicity of native collagen. FLS fibrils can be formed in vitro by addition of alpha(1)-acid glycoprotein to an acidified solution of monomeric collagen, followed by dialysis of the resulting mixture. We have investigated the ultrastructure of FLS fibrils formed in vitro using the atomic force microscope (AFM). The majority of the fibrils imaged showed typical diameters of approximately 150nm and had a distinct banding pattern with a approximately 250nm periodicity. However, we have also observed an additional type of FLS fibril, which is characterized by a secondary banding pattern surrounding the primary bands. These results are compared with those obtained in past investigations of FLS ultrastructure carried out using the transmission electron microscope (TEM). The importance of the fibril's surface topography in TEM staining patterns is discussed. Images of FLS fibrils in various stages of assembly have also been collected, and the implications of these images in determining the mechanism of assembly and the formation of the characteristic banding pattern of the fibrils is discussed. PMID- 11006516 TI - Ultrastructure and assembly of segmental long spacing collagen studied by atomic force microscopy. AB - The in vitro formation of segmental long spacing (SLS) collagen as induced by the addition of ATP to acidified Type I collagen solutions has been examined with the atomic force microscope (AFM). AFM images obtained suggest that the assembly proceeds in a stepwise manner, through an intermediate stage of oligomers, which then associate laterally to form the so-called "SLS crystallites". Attempts to induce SLS formation by the addition of other polyanionic species to monomeric collagen solutions met with mixed success; ATP-gamma-S and GTP produced SLS crystallites, whereas inorganic phosphate and other polyanionic dyes did not. This indicates that the formation of SLS cannot simply be attributed to the negation of positive charges believed to be located on the end of the collagen monomer, but rather it is a complex function of the structure and charge of both the collagen monomer and polyanion. PMID- 11006517 TI - Changing trends PMID- 11006518 TI - Targeting TGF could counter diabetic nephropathy. PMID- 11006519 TI - Kaposi's sarcoma-associated virus mounts novel attack on immune system. PMID- 11006520 TI - Vaccine against breast and ovarian cancer still on the horizon. PMID- 11006521 TI - Another piece in the hypertension puzzle. PMID- 11006522 TI - Nurture leads the race in the control of cancer. PMID- 11006523 TI - Communication genes clustered on 7q31. PMID- 11006524 TI - Memory cells in the immune system: how to be efficient, even in small numbers. PMID- 11006525 TI - Peptic ulcer irony. PMID- 11006526 TI - Ageing and the brain, a first glimpse at age-induced altered gene expression. PMID- 11006527 TI - Protein aggregation in Huntington's and Parkinson's disease: implications for therapy. AB - The accumulation of highly insoluble intracellular protein aggregates in neuronal inclusions is a hallmark of Huntington's disease (HD) and Parkinson's disease (PD) as well as several other late-onset neurodegenerative disorders. The aggregates formed in vitro and in vivo generally have a fibrillar morphology, consist of individual beta-strands and are resistant to proteolytic degradation. Although the causal relationship between aggregate formation and disease remains to be proven, the gradual deposition of mutant protein in neurons is consistent with the late-onset and progressive nature of symptoms. Recently, circumstantial evidence from mouse and Drosophila model systems suggests that abnormal protein folding and aggregation play a key role in the pathogenesis of both HD and PD. Therefore, a detailed understanding of the molecular mechanisms of protein aggregation and its effects on neuronal cell death could open new opportunities for therapy. PMID- 11006528 TI - Secretases as targets for the treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is the major cause of dementia in most developed countries. Treatment to modify this disease is currently unavailable, but needed urgently. The amyloid-cascade hypothesis proposes that amyloid beta-peptide (Abeta), found in the plaques characteristic of AD, plays an early, critical role in the disease process. It follows that preventing the generation of Abeta could be therapeutically useful in all cases of AD. Inhibition of the secretases that produce Abeta from a large precursor protein is the main approach to achieve this goal. PMID- 11006529 TI - RAS inhibitors: potential for cancer therapeutics. AB - As RAS oncoproteins play a major role in human malignancy, inhibiting RAS function is a promising approach for developing anticancer therapies. Among these approaches are agents such as farnesyltransferase inhibitors (FTIs) and the nontoxic farnesylcysteine analogue farnesylthiosalicylic acid (FTS) that dislodges all RAS isoforms from the membrane, as well as methods to restore regulation of RAS-GTP levels and to alter the interaction of RAS-GTP with downstream targets. PMID- 11006530 TI - X-linked severe combined immunodeficiency: from molecular cause to gene therapy within seven years. AB - X-linked severe combined immunodeficiency (XSCID) is the most common form of SCID. The discovery of the genetic defect in this disease, namely mutations in the gene encoding the common cytokine receptor gamma chain, gammac, was reported just over seven years ago. In the subsequent period, a tremendous amount of knowledge about the biology and function of this protein has been generated. Moreover, gammac-knockout mice have been generated and their immune systems successfully reconstituted by gene therapy. Furthermore, initial attempts at using gene therapy to treat patients with XSCID have been successful for more than ten months, making this disease perhaps the most promising to date for treatment with such a strategy. PMID- 11006531 TI - Murine models of malignant melanoma. PMID- 11006533 TI - Analytical techniques - from proteome analysis to mechanical phenotypes of multiprotein assemblies. Editorial overview PMID- 11006532 TI - Web alert. AB - A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Chemical Biology. PMID- 11006534 TI - Mass spectrometry and proteomics. AB - Proteomics is the systematic analysis of the proteins expressed by a cell or tissue, and mass spectrometry is its essential analytical tool. In the past two years, incremental advances in standard proteome technology have increased the speed of protein identification with higher levels of automation and sensitivity. Furthermore, new approaches have provided landmark advances in determining functionally relevant properties of proteins, including their quantity and involvement within protein complexes. PMID- 11006535 TI - Advances in multiple wavelength anomalous diffraction crystallography. AB - In only a few years, multiple wavelength anomalous diffraction (MAD) phasing has advanced from an esoteric technique used in only a few favorable cases to the method of choice for solving new macromolecular structures. Before 1994, MAD phasing had been used for fewer than a dozen new structure determinations. In 1999 alone, well over 100 new structures were determined by MAD phasing. The meteoric rise in MAD applications resulted from the availability of new synchrotron beamlines, equipped with low bandpass optics, fast readout detectors, cryogenic cooling and user-friendly interfaces. The power of MAD phasing has been amplified by the availability of new computer programs for locating the positions of the anomalous scattering atoms and for calculating phases from the experimental data. Phasing by anomalous scattering techniques has been applied to structures as large as 640 kDa and 120 selenium atoms in the asymmetric unit. The practical size limitation for application of MAD phasing techniques has not yet been encountered. PMID- 11006536 TI - Solid-state NMR as a probe of amyloid fibril structure. AB - Amyloid fibrils are intrinsically noncrystalline, insoluble, high-molecular weight aggregates of peptides and proteins, with considerable biomedical and biophysical significance. Solid-state NMR techniques are uniquely capable of providing high-resolution, site-specific structural constraints for amyloid fibrils, at the level of specific interatomic distances and torsion angles. So far, a relatively small number of solid-state NMR studies of amyloid fibrils have been reported. These have addressed issues about the supramolecular organization of beta-sheets in the fibrils and the peptide conformation in the fibrils, and have concentrated on the beta-amyloid peptide of Alzheimer's disease. Many additional applications of solid-state NMR to amyloid fibrils from a variety of sources are anticipated in the near future, as these systems are ideally suited for the technique and are of widespread current interest. PMID- 11006537 TI - Fluorescence resonance energy transfer as a structural tool for nucleic acids. AB - Fluorescence resonance energy transfer is a spectroscopic method that provides distance information on macromolecules in solution in the range 20-80 A. It is particularly suited to the analysis of the global structure of nucleic acids because the long-range distance information provides constraints when modelling these important structures. The application of fluorescence resonance energy transfer to nucleic acid structure has seen a resurgence of interest in the past decade, which continues to increase. An especially exciting development is the recent extension to single-molecule studies. PMID- 11006538 TI - Vibrational spectroscopy as a tool for probing protein function. AB - Vibrational spectroscopy has become increasingly important as a tool for understanding the mechanisms of photosystem II, phytochrome and terminal oxidases. More general enzymatic or receptor systems have been studied, opening a new field of applications. Femtosecond infrared pump/probe studies of the important amide-I band seem to provide a basis for its molecular and structural interpretation. PMID- 11006540 TI - Mechanisms. Mechanisms Of catalysis - macroscopic and microscopic progress PMID- 11006539 TI - Force spectroscopy with single bio-molecules. AB - For many biological molecules, force is an important functional and structural parameter. With the rapidly growing knowledge about the relation between structure, function, and force, single-molecule force spectroscopy has become a versatile analytical tool for the structural and functional investigation of single bio-molecules in their native environments. Within the past year, detailed insights into binding potentials of receptor ligand pairs, protein folding pathways, molecular motors, DNA mechanics and the functioning of DNA-binding agents (such as proteins and drugs), as well as the function of molecular motors, have been obtained. PMID- 11006542 TI - Computational enzymology. AB - Recent advances in computational methods and the availability of fast, affordable computers have made the modeling of enzymatic reactions practical. The remaining challenges include achieving the accuracy level at which thermodynamic parameters and kinetic constants for different substrates, mutant enzymes, or in the presence of allosteric effectors can be predicted quantitatively. PMID- 11006541 TI - Native disulfide bond formation in proteins. AB - Native disulfide bond formation is critical for the proper folding of many proteins. Recent studies using newly identified protein oxidants, folding catalysts, and mutant cells provide insight into the mechanism of oxidative protein folding in vivo. This insight promises new strategies for more efficient protein production. PMID- 11006543 TI - Cyclooxygenase mechanisms. AB - Several advances have occurred in the past year in our understanding of cyclooxygenase catalysis. The role of specific heme oxidation states in the formation of catalytically competent tyrosyl radicals has been defined; the identity of physiological hydroperoxide activators has been established; and the participation of individual amino acids in substrate binding and oxygenation has been elucidated. PMID- 11006544 TI - Ribonuclease P: a ribonucleoprotein enzyme. AB - The ribonucleoprotein ribonuclease P catalyzes the hydrolysis of a specific phosphodiester bond in precursor tRNA to form the mature 5' end of tRNA. Recent studies have shed light on the structures of RNase-P-RNA-P-protein and RNase-P RNA-precursor-tRNA complexes, as well as on the positions of catalytic metal ions, emphasizing the importance of the structure to the catalytic function. PMID- 11006545 TI - Nitrogenase: standing at the crossroads. AB - Nitrogenase catalyzes the ATP-dependent reduction of dinitrogen to ammonia, which is central to the process of biological nitrogen fixation. Recent progress towards establishing the mechanism of action of this complex metalloenzyme reflects the contributions of a combination of structural, biochemical, spectroscopic, synthetic and theoretical approaches to a challenging problem with implications for a range of biochemical and chemical systems. PMID- 11006546 TI - Modular evolution of the purine biosynthetic pathway. AB - Structural studies, sequence alignments, and biochemistry have provided new insights into the evolution of the purine biosynthetic pathway. The importance of chemistry, the binding of ribose 5-phosphate (common to all purine biosynthetic intermediates), and transient protein-protein interactions in channeling of chemically unstable intermediates have all been examined in the past few years. PMID- 11006547 TI - Glycosidase mechanisms. AB - Insights into glycosidase mechanisms have come from X-ray crystallographic studies on complexes with substrate analogs and inhibitors, representing all the intermediate species along the reaction coordinate. Site-directed mutagenesis continues to play a significant role in understanding mechanisms, but is also proving important in generating glycosidases of modified mechanism or specificity. PMID- 11006548 TI - Mechanisms of DNA replication. AB - Advances in recent years have led to exciting new ideas about the initiation, regulation and coordination of DNA replication. Structural studies have yielded fascinating glimpses of replisome action. In addition, the involvement of replication proteins in other cellular processes has blurred the lines between replication, repair and recombination. PMID- 11006549 TI - Uequiv: its past, present and future. AB - The reporting of U(equiv) and its standard uncertainty has a chequered history. In spite of the recommendation of the IUCr Commission on Journals that authors use the definition of U(equiv) of their own choice, possibly without standard uncertainties, there still seems to be some confusion amongst referees and editors about the status of this derived parameter. It is shown that neither of the common definitions are very useful, and that the standard uncertainty computed from the refinement normal matrix is almost worthless. A potential alternative derived parameter is proposed. PMID- 11006550 TI - Architecture of crystal structures from square planes. AB - The crystal structures of ordered b.c.c. (body-centered cubic), f.c. c. (face centered cubic) or primitive cubic alloys AxBy and related NaCl, ZnS or CaF(2) derivative structures are characterized by the self-coordination numbers T(1), T(2) of the A atoms with A atoms. Structures with identical T(1) and T(2) values for all A atoms are at the corners of T(1) and T(2) structure maps, and can be analyzed for attractive or repulsive interactions of A atoms. Most observed structures are at the borders of the structure map and can be obtained by approximately 10 different combinations of structural units. The different combination mechanisms explain e.g. the shear structures of CuAu II or Nb(2)O(5) and the occurrence of vacancies in NaCl-related structures like NbO. PMID- 11006551 TI - Refinement of framework disorder in dehydrated CaA zeolite from single-crystal synchrotron data. AB - An accurate knowledge of zeolite structure is required for understanding their selective sorption capacities and their catalytic properties. In particular, the positions of the exchangeable cations and their interactions with the framework are essential. The present study deals with the accurate crystal structure determination of a fully exchanged and fully dehydrated CaA zeolite (Ca(48)Al(96)Si(96)O(384), Fm3c, a = 24.47 A) using single-crystal high resolution synchrotron X-ray diffraction [(sin straight theta/lambda)(max) = 1.4 A(-1)]. It is shown that cation exchange severely distorts the skeleton, especially around the O2 atom. The high-resolution synchrotron data reveal that this latter O atom is disordered and lies out of the mirror plane it occupies in other A-type zeolites. This feature is related to that observed for Ca(2+) cations. PMID- 11006552 TI - X-ray diffraction study of K3NdSi7O17: a new framework silicate with a linear Si O-Si bond. AB - Hydrothermal investigations in the high-silica region of the K(2)O-Nd(2)O(3) SiO(2) system, carried out in a search for novel fast-ion conductors (FICs), yielded the new compound tripotassium neodymium heptasilicate, K(3)NdSi(7)O(17). Single-crystal X-ray methods revealed that K(3)NdSi(7)O(17) crystallizes in space group P3, has lattice constants a = 16.131 (2) and c = 7.7146 (19) A, Z = 4, and 22 atoms in the asymmetric unit. Refinement was carried out to a residual, R(F), of 0.0253 and a weighted residual, wR(F(2)), of 0.0702 using anisotropic displacement parameters for all atoms. The silicate anion forms an interrupted framework, within which both Nd octahedra and K polyhedra are situated. The structure is unusual in that it contains a symmetry-constrained Si-O-Si bond angle of 180 degrees. No isomorphs to K(3)NdSi(7)O(17) are known. PMID- 11006553 TI - Twinning and defects in N-Nb2O5. AB - N-Nb(2)O(5) [C2/m (No. 12), a = 28.51, b = 3.830 and c = 17.48 A, and beta = 124.8 degrees ] has been investigated by means of selected-area electron diffraction (SAED) and high-resolution transmission electron microscopy (HRTEM). N-Nb(2)O(5) is domain twinned, with the twin plane perpendicular to the c* axis. The domains are rather small and the domain twinning can sometimes be best explained as stacking faults. A second type of coherent twinning at an angle of 90 degrees to the other two domain directions was also found. These domains are linked together by areas containing blocks of different sizes, similar to the disordered block arrangement observed in M-Nb(2)O(5). PMID- 11006554 TI - Synchrotron X-ray analysis of RbTiOAsO4. AB - Structure factors for rubidium oxotitanium arsenate, RbTiOAsO(4), were measured at 293 K with focused synchrotron X-radiation [lambda = 0.7500 (9) A] using a fast avalanche photodiode counter. The accurate synchrotron single-crystal data are of sufficient quality and resolution to detect the splitting positions of the Rb cations at room temperature. Strong accumulation of the Deltarho difference charge density near the Rb atoms at a distance of approximately 0.5 A in the -c direction can be attributed to the partial occupancy of additional sites related by pseudosymmetry. This type of static and/or dynamic disorder is temperature dependent and seems to be universal for the KTiOPO(4) family of compounds. The best modelling of the experimental data was obtained with the Rb atom in split positions described within the harmonic approximation and the multipole functions for the other atoms. The Deltarho density features in the Ti-O and As-O covalent bonds can be related to the linear and non-linear susceptibility using bond polarization theory. The charge-density maps reflect the anisotropy of non-linear susceptibility, which is larger for directions where locally antisymmetric components of Deltarho are strong. PMID- 11006555 TI - Systematic prediction of new ferroelectrics in space group P3. AB - The current release of the Inorganic Crystal Structure Database contains a total of 57 entries under space group P3 that correspond to 50 different materials. There are 21 structures reported with this space group that satisfy the criteria for ferroelectricity, at a confidence level that depends on the reliability of the underlying structural determination. One ferroelectric discovered earlier is also listed. In addition, the database contains 19 entries that probably should be assigned to a centrosymmetric space group, seven that are polar but probably not ferroelectric and two that are without atomic coordinates. Seven entries are either duplicates or present additional structural studies of the same material. Structures in space group P3 identified as potentially new ferroelectrics include LiAsCu(0.93), Na(2)UF(6), BiTeI, BaGe(4)O(9), alpha-UMo(2)O(8), Cu(2)SiS(3), Co(IO(3))(2), Sr(7)Al(12)O(25), KSn(2)F(5), YbIn(2)S(4), Na(5)CrF(2)(PO(4))(2), Sn(ClO(2))(2)(ClO(4))(6), Eu(3)BWO(9), Li(H(2)O)(4)B(OH)(4).2H(2)O, Mn(3)V(1/2)(SiO(4))O(OH)(2), Ca(6)(Si(2)O(7))(OH)(6), Na(6. 9(2))[Al(5.6(1))Si(6.4(1))O(24)](S(2)O(3))(1.0(1)).2H(2)O, BaCa(2)In(6)O(12), Ni(H(2)O)(6)[Sb(OH)(6)](2), Sr(4)Cr(3)O(9) and Cu(5)O(2)(VO(4))(2).CuCl(2). PMID- 11006556 TI - Crystal structure and charge distribution of YbFeMnO4. AB - The structure of synthetic YbFeMnO(4) has been refined by single-crystal X-ray diffraction. Space group R3m, a = 3.4580 (1), c = 25.647 (3) A, V = 265.59 (3) A(3), Z = 3. Yb is in octahedral coordination, whereas Fe and Mn are disordered on a single crystallographic type of trigonal bipyramid, in which the cation is off-centred from the basal plane. Assuming perfect stoichiometry, R(1) = 0.0195, but the charge distribution (CD) analysis suggests incomplete occupation of the Yb site. Refinement of the occupancy lowers R(1) to 0.0175, resulting in s.o.f.(Yb) = 0. 963 (3), with a significant improvement of the Fourier difference. The electroneutrality is likely preserved through incomplete occupancy of one of the two oxygen sites: the compound is thus non stoichiometric, with the formula Yb(0.963)FeMnO(3.945). Another mechanism for preserving the electroneutrality is the oxidation of a small amount of Mn(2+) to Mn(3+), which is, however, less probable because of the reduction conditions in which the sample was synthesized. Both models give a satisfactorily CD result, but they cannot be definitively distinguished by X-ray data. PMID- 11006557 TI - Two-dimensional incommensurately modulated structure of (Sr0.13Ca0. 87)2CoSi2O7 crystals. AB - The incommensurate structure of (Sr(0.13)Ca(0.87))(2)CoSi(2)O(7) at room temperature has been determined from single-crystal X-ray diffraction data. The compound has a non-centrosymmetric tetragonal basic cell of a = 7.8743 (4) and c = 5.0417 (2) A with the space group P4;2(1)m. The refinements of the basic structure converged to R = 0.038 for 757 main reflections. The two-dimensional incommensurate structure is characterized by the wavevectors q(1) = 0.286 (3)(a* + b*) and q(2) = 0.286 (3)(-a* + b*), where a*, b* are the reciprocal lattice vectors of the basic structure. With the (3 + 2)-dimensional superspace group P(p4mg)(P4;2(1)(m)), the refinements converged to R = 0.071 for 1697 observed reflections (757 main and 940 satellite reflections). The structure is described in terms of displacement of the atoms, rotation, distortion of CoO(4) and SiO(4) tetrahedra, and the partial ordering of the Sr and Ca atoms accompanied with the modulation. Correlated evolution of these features throughout the crystal gives rise to various oxygen coordination around Ca/Sr. Comparison of the derived modulated structure to that of Ca(2)CoSi(2)O(7) clarified that the partial substitution of Ca by large alkaline-earth atoms such as Sr should decrease the distortion of the polyhedra around the cations. PMID- 11006558 TI - Isomorphism and pseudosymmetry in 2,6-dichloro- and 2, 6-dibromobenzonitrile. AB - The title compounds are isomorphous, with two half-molecules in the asymmetric unit of a monoclinic unit cell in the space group C2/m. For the chloro compound: a = 18.0525 (8), b = 20.7374 (10), c = 3. 8334 (2) A, beta = 101.143 (1) degrees; for the bromo compound: a = 18.2269 (15), b = 21.349 (2), c = 3.9663 (3) A, beta = 101.446 (2) degrees. The dominant intermolecular interactions are between halogen atoms rather than between halogen and nitrile, as is the case in the corresponding 2,4,6-trihalobenzonitriles. The molecules pack into layers with a fourfold pseudosymmetry. The pseudosymmetry appears to be the consequence of halogen-halogen intermolecular interactions. PMID- 11006559 TI - Two-dimensional packing and pseudosymmetry. AB - It is possible to understand, or at least rationalize, the occurrence of pseusosymmetry elements in some crystals as a consequence of a higher symmetry in a two-dimensional layer being obscured by the three-dimensional packing of the layers. Six examples, including both simple compounds and co-crystals and involving six different symmetries, are described. PMID- 11006560 TI - Packing modes of (R,R)-tartaric acid esters and amides. AB - The molecular packing modes of a series of mono- and diamides of (R, R)-tartaric acid are discussed on the basis of their crystal structures. Derivatives include combinations of methylester, amide, N-methylamide and N,N-dimethylamide groups, both symmetrically and asymmetrically substituted. The symmetrically substituted derivatives do not utilize their C(2) symmetry in the crystal. The packing of primary tartramides seems to be driven by NH.O=C hydrogen bonds and supplemented by strong OH.O=C and weak NH.OH bonds. On the other hand, in derivatives containing methylester and/or methylamide groups OH.OH.O=C hydrogen-bond patterns seem to dominate. Types of aggregates, characteristic for the investigated derivatives, include cyclic dimers and ring systems analogous to the dimers, but formed by two different although complementary functional groups, as well as sets of chains aligned in a manner resembling the helical arrangement of peptides. The helices are formed along the screw axis with an identity period of approximately 5 A. In tartramic acids, containing in one molecule both carboxyl and amide functions, in competition between the two groups to control the molecular arrangement, the latter dominates, unless it is N-substituted tartramide, in which case the carboxyl group predominates. Problems with packing, which occur in some of the structures owing to the steric bulk of the methyl groups, are overcome by changes in conformation (esters) or by co-crystallization with solvent water molecules (methylamides and dimethylamides). These derivatives are also more likely to crystallize with multiple asymmetric units. PMID- 11006561 TI - Intramolecular hydrogen bonds: common motifs, probabilities of formation and implications for supramolecular organization. AB - A systematic survey of the Cambridge Structural Database (CSD) has identified all intramolecular hydrogen-bonded ring motifs comprising less than 20 atoms with N and O donors and acceptors. The probabilities of formation P(m) of the 50 most common motifs, which chiefly comprise five- and six-membered rings, have been derived by considering the number of intramolecular motifs which could possibly form. The most probable motifs (P(m) > 85%) are planar conjugated six-membered rings with a propensity for resonance-assisted hydrogen bonding and these form the shortest contacts, whilst saturated six-membered rings typically have P(m) < 10%. The influence of intramolecular-motif formation on intermolecular hydrogen bond formation has been assessed for a planar conjugated model substructure, showing that a donor-H is considerably less likely to form an intermolecular bond if it forms an intramolecular one. On the other hand, the involvement of a carbonyl acceptor in an intramolecular bond does not significantly affect its ability to act as an intermolecular acceptor and thus carbonyl acceptors display a substantially higher inclination for bifurcation if one hydrogen bond is intramolecular. PMID- 11006562 TI - Graph-set and packing analysis of hydrogen-bonded networks in polyamide structures in the Cambridge Structural Database. AB - The hydrogen-bond networks and crystal packing of 81 unique secondary di- and polyamides in the Cambridge Structural Database are investigated. Graph-set analysis, as implemented in the RPluto program, is used to classify network motifs. These have been rationalized in terms of the relative dispositions of the amide groups. Peptide and retropeptides exhibit significant conformational flexibility, which permits alternative hydrogen-bonding patterns. In peptides, dihedral angles of -psi approximately varphi approximately 105 degrees allow an antiparallel ladder arrangement, containing rings of either the same or alternating sizes. For retropeptides, and diamides with an odd number of CH(2) spacers, this conformation leads to a parallel ladder with rings of equal size. If varphi approaches -60 degrees and psi 180 degrees, ladders adopt a helical twist, and if the conformation is distorted further, a three-dimensional network is usually adopted. Diamides with aromatic or an even number of CH(2) spacers generally form either antiparallel ladders or sheets, although some exhibit both polymorphs. Symmetry relationships within and between hydrogen-bonded chains, ladders and sheets in the crystal packing have also been analysed. Polyamides form considerably more complex networks, although many of the structural motifs present in the diamides occur as components of these networks. PMID- 11006563 TI - Conformational transformation coupled with the order-disorder phase transition in 2-methyl-1,3-cyclohexanedione crystals. AB - The half-chair conformation of the dynamically disordered molecular ring of 2 methyl-1,3-cyclohexanedione, C(7)H(10)O(2), transforms to a sofa below Tc = 244 K, when the crystal undergoes a continuous phase transition induced by the onset of halting large-amplitude vibrations of methylene groups C(4)H(2) and C(5)H(2). The temperature dependence of the crystal structure has been investigated by X ray diffraction. The Ibam symmetry of the crystal reduces below Tc to space group Pccn. The mechanism of the phase transition and of the conversion of the ring conformation is discussed. PMID- 11006564 TI - N-(6-Amino-3,4-dihydro-3-methyl-5-nitroso-4-oxopyrimidin-2-yl) derivatives of glycine, valine, serine, threonine and methionine: interplay of molecular, molecular-electronic and supramolecular structures. AB - In each of N-(6-amino-3, 4-dihydro-3-methyl-5-nitroso-4-oxopyrimidin-2-yl)valine, C(10)H(15)N(5)O(4) (3) (orthorhombic, P2(1)2(1)2(1)), N-(6-amino-3, 4-dihydro-3 methyl-5-nitroso-4-oxopyrimidin-2-yl)serine monohydrate, C(8)H(11)N(5)O(5).H(2)O (4) (orthorhombic, P2(1)2(1)2(1)), and N-(6-amino-3, 4-dihydro-3-methyl-5-nitroso 4-oxopyrimidin-2-yl)threonine, C(9)H(13)N(5)O(5)(5) (monoclinic, P2(1)), the C nitroso fragments exhibit almost equal C-N and N-O bond lengths: the C-N range is 1. 315 (3)-1.329 (3) A and the N-O range is 1.293 (3)-1.326 (3) A. In each compound there are also very short intermolecular O-H.O hydrogen bonds, in which carboxyl groups act as hydrogen-bond donors to the nitrosyl O atoms: the O.O distances range from 2.440 (2) to 2. 504 (4) A and the O-H.O angles lie between 161 and 163 degrees. An interpretation of the relationship between the unusual intramolecular bond lengths and the very short intermolecular hydrogen bonds has been developed based on database analysis and computational modelling. In each of (3)-(5) there is an extensive network of intermolecular hydrogen bonds, generating three-dimensional frameworks in (3) and (5), and two-dimensional sheets in (4). PMID- 11006565 TI - Conformational preferences and supramolecular aggregation in 2 nitrophenylthiolates: disulfides and thiosulfonates. AB - In each of the asymmetrically substituted disulfides 2-nitro-4'-methyldiphenyl disulfide, C(13)H(11)NO(2)S(2) (1), 2-nitro-4'-chlorodiphenyl disulfide, C(12)H(8)ClNO(2)S(2) (2), 2, 4-dinitro-4'-methyldiphenyl disulfide, C(13)H(10)N(2)O(4)S(2) (3), and 2,4-dinitrophenyl-2'-methoxycarbonylmethyl disulfide, C(9)H(8)N(2)O(6)S(2) (4), and in both of the symmetrically substituted disulfides bis(2-nitrophenyl) disulfide, C(12)H(8)N(2)O(4)S(2) (5), and bis(2 nitro-4-trifluoromethylphenyl) disulfide, C(14)H(6)F(6)N(2)O(4)S(2) (6), the 2 nitro groups are essentially coplanar with the adjacent aryl ring and the S atom remote from the nitrated aryl ring is also essentially coplanar and transoid to the nitro group. In S-(2-nitrophenyl) 2-nitrobenzene thiosulfonate, C(12)H(8)N(2)O(6)S(2) (7), which contains three independent molecules in the asymmetric unit, all six of the independent nitro groups are twisted out of the plane of the adjacent aryl rings. The crystal structures of (1)-(3) contain isolated molecules, that of (4) contains centrosymmetric dimers held together by C-H.O hydrogen bonds, while in the structures of (5)-(7), respectively, the C-H.O hydrogen bonds generate one-, two- and three-dimensional arrays. The interplay between molecular conformation and supramolecular aggregation is discussed. PMID- 11006566 TI - Ferroelastic structures of n-pentyl-, n- hexyl- and n-nonylammonium dihydrogenphosphate crystals. AB - This study reports the structure redeterminations of C(5)H(11)NH(3)(+).H(2)PO(4)( ) (n-pentylammonium dihydrogenphosphate, C5ADP), C(6)H(13)NH(3)(+).H(2)PO(4)(-) (n-hexylammonium dihydrogenphosphate, C6ADP) and C(9)H(19)NH(3)(+).H(2)PO(4)(-) (n-nonylammonium dihydrogenphosphate, C9ADP). The structures are monoclinic (P2(1)/n), belonging to the series of previously studied structures C2ADP-C8ADP and C10ADP. The structures exhibit reproducible ferroelastic switching. There are hydrogen bonds between the dihydrogenphosphates and the n-alkylammonium groups. Among them there are two hydrogen bonds with hydrogens which hop from the donor to the acceptor oxygens during the ferroelastic switching. C5ADP as well as C3ADP differ from the other members of the series by packing of the double layers of the dihydrogenphosphates. Moreover, the packing of n-alkylammonium molecules in all these structures depends on the parity of the number of atoms in the n alkylammonium chains. All the samples contained two domains and their structures were refined as twins. PMID- 11006567 TI - Orthorhombic tau-Zn-Mg-Dy phase related to a Frank-Kasper type decagonal quasicrystal. AB - We report a novel Zn-Mg-Dy phase, denoted as the tau-phase, whose structure is related to that of the Frank-Kasper type decagonal Zn-Mg-Dy phase, based on electron diffraction and high-resolution transmission electron microscopy studies. A plausible atomic model is proposed, which is constructed by two types of atomic clusters related to those in the Zn(7)Mg(4) crystalline compound. PMID- 11006568 TI - X-ray structure of barium titanate - missed opportunities. AB - Anomalous X-ray scattering effects are quite extensive in the noncentrosymmetric ferroelectric structure of barium titanate, and typical estimates for three published X-ray diffraction experiments are computed. These data show that the Bijvoet pairs should not be averaged before least-squares refinement for this polar crystal with small atomic displacements from a higher symmetric space group. PMID- 11006573 TI - Changes in serum and tissue zinc levels in sex hormone-induced prostatic carcinogenesis in the noble rat. AB - We investigated the changes in serum and tissue zinc levels in the Noble rat prostate gland under different pathological conditions induced by the administration of a combination of testosterone and 17beta-estradiol. The results showed that there were significant differences in serum zinc values between normal and hormone-treated rats with prostatic hyperplasia, dysplasia and prostatic carcinoma (p < 0.05), although there was no significant difference among rats with different forms of prostatic lesions (i.e. hyperplasia, dysplasia and prostatic adenocarcinoma). There was also a difference in zinc content between the lateral prostate (LP), ventral prostate (VP) and dorsal prostate (DP) in normal rats. The zinc levels of LP were several times greater than those of either VP or DP (p < 0.01). There was also a great difference in zinc levels between the normal and the hyperplastic/dysplastic and carcinomatous LP and VP (p < 0. 05). The levels of zinc in both LP and VP were increased in hyperplasia/dysplasia and carcinoma. On the other hand, the zinc levels of LP were much higher than those of VP in hyperplasia/dysplasia and carcinoma (p < 0.01), which may be correlated with the incidence of prostate cancers in these lobes (i. e. higher in LP and much lower in VP). In contrast, in DP, the levels of zinc were not affected, which may be correlated with the very low incidence of carcinoma in this lobe. Our data suggest that the difference in zinc levels among these lobes reflect the heterogeneity in zinc content in various lobes of the rat prostate. The growth and development of prostatic lesions in LP and VP may be positively correlated with the significant increase in tissue zinc levels in these lobes. On the other hand, the lack of response of DP to carcinogenesis may be due to its relatively stable low zinc levels. It is suggested that tissue zinc content may be used as a marker for prostatic lesions, including hyperplasia, dysplasia and carcinoma, while serum zinc levels may be a useful indicator for abnormal prostatic growth. PMID- 11006572 TI - Ascitic growth of a spontaneous transplantable T cell lymphoma induces thymic involution. 2. Induction of apoptosis in thymocytes. AB - It has been observed that the progressive ascitic growth of a transplantable T cell lymphoma of spontaneous origin, designated as Dalton's lymphoma (DL), induces inhibition of various immune responses and is associated with an involution of the thymus accompanied by a massive depletion of the cortical region and alteration in the distribution of thymocytes, with a decrease in CD4+CD8+, CD4+CD8- and CD4-CD8+ thymocytes. Morphological evaluation of thymocytes from DL-bearing mice revealed that with the progression of DL, a majority of thymocytes exhibited morphological features characteristic of apoptotic cell death, which included contracted cell bodies, condensed, uniformly circumscribed and densely stained chromatin, and membrane-bound apoptotic bodies containing one or more nuclear fragments. Quantitative and qualitative analysis of the DNA extracted from the thymocytes of DL-bearing mice revealed DNA fragmentation that increased concomitantly with the progression of DL and showed an oligonucleosomal DNA ladder pattern upon agarose gel electrophoresis, a hallmark of apoptotic cell death. Attempts to identify apoptotic factor(s) showed that the serum of DL-bearing mice contained certain soluble factor(s) that augmented the induction of apoptotis in thymocytes in a time- and dose-dependent manner. Although DL cells or their products, such as DL-cell-conditioned medium or DL-cell-free ascitic fluid, could also induce apoptosis of thymocytes in vitro, the magnitude of the same was consistently lower than that induced by the serum of DL-bearing mice. Further, elucidation of the mechanism of apoptosis induction in thymocytes with respect to the involvement of apoptosis-related genes revealed that the death pathway followed an interleukin-1 beta-converting enzyme-dependent, Fas-mediated apoptotic cascade, with a concomitant increase in the protein products of the bax, bad, p53, fas and fasL genes and cleavage of the 23-kD N-terminal fragment of Bcl-2 that exhibited Bax-like death effector properties. PMID- 11006574 TI - Synergistic cytotoxic interactions between sodium butyrate, MG132 and camptothecin in human retinoblastoma Y79 cells. AB - This paper studies the effects caused in human retinoblastoma Y79 cells by treatment with combinations of sodium butyrate, the inhibitor of topoisomerase I camptothecin and the inhibitor of 26S proteasome MG132. The combination of sodium butyrate and camptothecin resulted in a strong synergistic cytotoxicity, as revealed by combination indices of 0.77 and 0.52 calculated at IC(50) and IC(75). Synergistic interactions were also demonstrated for combinations of sodium butyrate and MG132, camptothecin and MG132 and for a combination of all three compounds. The cytotoxic effects observed after the combined treatments can be considered a consequence of apoptosis, as suggested by the appearance of morphological signals of apoptosis and by the activation of caspase-3 with degradation of poly-ADP ribose polymerase and lamin B. Treatment of Y79 cells with sodium butyrate alone lowered the levels of p53, E2F-1 and Bcl-2. The addition of MG132 to sodium butyrate counteracted the effect on p53 only, while the addition of camptothecin to sodium butyrate counteracted the effect on both p53 and E2F-1. The treatment of Y79 cells with the triple combination increased the level of p53, decreased that of Bcl-2, while the level of E2F-1 was not modified. We suggest that the effects exerted on the levels of these regulatory proteins can explain the synergistic interactions demonstrated between sodium butyrate, camptothecin and MG132. PMID- 11006575 TI - Syndecan-1-dependent homotypic cell adhesion in HT58 lymphoma cells. AB - OBJECTIVES: Many cellular functions are controlled by cell-cell and cell-matrix interactions. It has recently been found that syndecans, transmembrane heparan sulphate (HS) proteoglycans, can act as receptors or co-receptors and modulate cell adhesion. Our aim was to study the role of syndecan-1 in the aggregation of human lymphoma cells, and to investigate its effect on cell survival. METHODS: Immunocytochemistry, confocal laser scanning microscopy, flow cytometry and aggregation/reaggregation bio-assays were used on HT58, BL41/95 and Raji lymphoma cell lines. RESULTS: Bio-assays showed that the aggregation of HT58 cells was inhibited by heparin, HS, removal of the HS chain and binding of the anti syndecan-1 monoclonal antibody. In the search for a counter-receptor of syndecan 1, several adhesion molecules were tested, but none of them proved to be the adhesion partner. In the case of heparitinase/trypsin digestion with long-term inhibition of HS synthesis (sodium chlorate treatment), the inhibited aggregation was accompanied by cell cycle arrest and the induction of apoptosis. CONCLUSIONS: The results obtained showed that surface syndecan-1 expression contributes to homotypic adhesion. In addition, HS chains, including those on syndecan-1, take part in the regulation of cell proliferation and active cell death in HT58 lymphoma cells. PMID- 11006576 TI - Epidermal growth factor receptor immunostaining and epidermal growth factor receptor-tyrosine kinase activity in proliferative and neoplastic human endometrium. AB - Epidermal growth factor receptor (EGFR) expression and EGFR-tyrosine kinase (EGFR TK) activity were measured in proliferative (n = 12) and neoplastic (n = 31) human endometrium. Immunoreactivity of EGFR was related to clinicopathological features, estrogen receptor (ER) and progesterone receptor (PR) status, and patient outcome. All proliferative and 27 neoplastic specimens expressed the EGFR. Expression of the EGFR was higher in proliferative endometrium than in endometrial cancer (p < 0.0001). ER immunostaining was observed in 19 of the endometrial carcinomas, while PR expression was demonstrated in only 12 neoplastic specimens. EGFR expression was not related to the ER/PR immunostaining in endometrial carcinomas. Clinicopathological features (age, stage, histological type, grade or depth of invasion) and clinical outcome were unrelated to EGFR immunoreactivity. EGFR-TK activity was detected in 29 of 31 endometrial neoplasms with a 9 times higher mean activity in neoplastic than in proliferative endometrial specimens. There was no relationship between the EGFR-TK activity and EGFR immunostaining in human neoplastic endometrium (p = 0.77). A trend towards a poorer outcome of patients with the EGFR-TK activity above 40 pmol/min/mg was observed, but was not statistically significant. These results support the view that the EGFR expression is downregulated in endometrial carcinomas compared to proliferative endometrial specimens. PMID- 11006577 TI - Antitumor activity of alpha fetoprotein and epidermal growth factor conjugates in vitro and in vivo. AB - Conjugates of carminomycin (Cm) with alpha-fetoprotein (AFP) and epidermal growth factor (EGF) were prepared and their cytotoxic activities were studied in vitro. Both conjugates showed cytotoxic activity which exceeded that of free Cm in tumor cell cultures of MCF-7, SKOV3, QOS, P388 and B16 cells. The antitumor effects of the conjugates were studied in vivo in mice with subcutaneous tumors of B16 and P388 cells. The Cm-AFP and Cm-EGF conjugates inhibited tumor growth and noticeably increased the mean life span in experimental animals. Our results suggest that the therapeutic activity of Cm can be significantly enhanced by conjugation to AFP or EGF. PMID- 11006578 TI - Clinical value of tumor markers for early detection of recurrence in patients with cervical adenocarcinoma and adenosquamous carcinoma. AB - OBJECTIVE: The clinical value of tumor markers for early detection of recurrence was investigated in 32 patients with cervical adenocarcinoma or adenosquamous carcinoma who had recurrent tumors. METHODS: Serum levels of CA 125, CA 19-9, carcinoembryonic antigen (CEA), and squamous cell carcinoma antigen (SCC), in addition to clinical status at the time of recurrence were investigated. RESULTS: Among the 32 patients, 26 had no symptoms at the time of recurrence. In 20 patients, elevated serum levels of tumor markers were the first sign of recurrence. In 21 patients with recurrent adenocarcinoma, the positive rates were 14% (CA 125), 62% (CA 19-9), 29% (CEA), and 5% (SCC). There were 71% of cases positive for CA 19-9 and/or CEA. In 11 patients with recurrent adenosquamous carcinoma, the corresponding positive rates were 37% (CA 125), 46% (CA 19-9), 64% (CEA), and 55% (SCC), with 100% positive for CA 19-9, CEA, and/or SCC. CONCLUSIONS: The combination of CA 19-9 and CEA is probably the most promising for detection of recurrent cervical adenocarcinoma. For adenosquamous carcinoma, the additional use of SCC is recommended. PMID- 11006579 TI - Determination of brain and plasma drug concentrations by liquid chromatography/tandem mass spectrometry. AB - A method is described for the evaluation of drug concentrations in plasma and brain from treated rats. The analyte is recovered from plasma or brain homogenate by liquid-liquid extraction and subsequently analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). A simple experimental protocol renders the procedure valuable for obtaining information rapidly on brain penetration and plasma exposure of specific classes of compounds. This methodology has been applied to evaluate brain penetration with 30 different compounds from the same discovery program. In an attempt to increase throughput in our screening efforts, mixture dosing was evaluated. Results from single compound administration were compared with results following administration of a mixture of four compounds. Preliminary results, with specific classes of compounds, show no major differences (ranking order) in brain or plasma concentrations between mixture dosing and single compound administration, suggesting that mixture dosing could be applicable to brain penetration studies in the drug discovery phase. PMID- 11006580 TI - Evaluation of the metal binding properties of the histidine-rich antimicrobial peptides histatin 3 and 5 by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) was used to investigate metal ion interactions with salivary peptides histatin 3 (H3) and histatin 5 (H5). Conformational changes of these peptides in the presence of metal ions were studied using circular dichroism spectroscopy. H3 and H5 formed high affinity complexes with Cu(2+) and Ni(2+) and, to a lesser extent, with Zn(2+). Both peptides show the potential for multiple binding sites for Cu(2+) and Ni(2+) and only a single strong binding site for Zn(2+). The binding of a third Cu(2+) ion to H3 seems to enable the binding of a fourth ion to H3. The binding of a second and third Ni(2+) ion to H5 has a similar effect in enabling the binding of a fourth ion. None of the metal ions examined stabilized a regular secondary structure for either peptide. Subtle changes in overall conformation are seen with the addition of Cu(2+) to both H3 and H5. PMID- 11006581 TI - Regioselective synthesis and electron impact mass spectral fragmentation differentiation of 2- or 3-isopropylthio, phenylthiosuccinic acid 4 methyl/isopropyl ethers. AB - 2- and 3-isopropylthio/phenylthiosuccinic acid 4-methyl/isopropyl esters were synthesized regioselectively by Michael additions of isopropylthiol/thiophenol to maleic anhydride, followed by alcoholysis with methanol/isopropanol. Their mass spectrometric behavior has been studied with the aid of mass-analyzed ion kinetic energy spectrometry and accurate mass measurements under electron impact ionization. All compounds show a tendency to eliminate an alkoxy group, an alcohol, or an alkoxycarbonyl group from the respective molecular ion. Some molecular ions also show a tendency to eliminate a molecule of water, propene or CO(2). The [M - MeCH = CH(2)](+) ions could further lose H(2)O to form substituted succinic anhydride ions, or lose CO(2) to form 3-substituted propionic acid ions. Both of these ions could further yield other small fragment ions by loss of CO(2)H, CO or other small fragments. It has been found that 2 isopropylthio- and 3-phenylthiosuccinic acid 4-esters show more abundant [M - H(2)O](+) peaks than their 3-isopropylthio and 2-phenylthio isomers in their mass spectra. PMID- 11006582 TI - A new method for the accurate determination of the isotopic state of single amide hydrogens within peptides using Fourier transform ion cyclotron resonance mass spectrometry. AB - A new method is presented to accurately determine the probability of having a deuterium or hydrogen atom on a specific amide position within a peptide after deuterium/hydrogen (D/H) exchange in solution. Amide hydrogen exchange has been proven to be a sensitive probe for studying protein structures and structural dynamics. At the same time, mass spectrometry in combination with physical fragmentation methods is commonly used to sequence proteins based on an amino acid residue specific mass analysis. In the present study it is demonstrated that the isotopic patterns of a series of peptide fragment ions obtained with capillary-skimmer dissociation, as observed with a 9.4 T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer, can be used to calculate the isotopic state of specific amide hydrogens. This calculation is based on the experimentally observed isotopic patterns of two consecutive fragments and on the isotopic binomial distributions of the atoms in the residue constituting the difference between these two consecutive fragments. The applicability of the method is demonstrated by following the sequence-specific D/H exchange rate in solution of single amide hydrogens within some peptides. PMID- 11006583 TI - The need for chromatographic and mass resolution in liquid chromatography/tandem mass spectrometric methods used for quantitation of lactones and corresponding hydroxy acids in biological samples. AB - Because of the potential in-source conversion between a lactone and the corresponding hydroxy acid, it has been recognized that a liquid chromatography/tandem mass spectrometric (LC/MS/MS) method developed for quantitation of a lactone drug in the presence of its hydroxy acid metabolite (or vice versa) must incorporate chromatographic separation between the two compounds, unless in-source conversion between the two compounds has been eliminated by the appropriate selection of the LC/MS/MS parameters. We now report that chromatographic separation between a lactone and its hydroxy acid will be required under certain LC/MS/MS conditions used even in the absence of in-source conversion. This is due to the fact that the 18-mass-unit difference between a lactone and its hydroxy acid is, by coincidence, different by only one mass unit from the 17-mass-unit difference between the [M + H](+) and [M + NH(4)](+) ions of the lactone or the hydroxy acid. Thus, the [M + H](+) ion of a hydroxy acid is higher than the [M + NH(4)](+) ion of its lactone by only one mass unit. Therefore, in a method developed for quantitation of a hydroxy acid drug utilizing a selected-ion-monitoring (SRM) scheme that incorporates its [M + H](+) ion as the precursor ion, the quantitation would be inaccurate due to the interference by the contribution of the A + 1 isotope response from the [M + NH(4)](+) ion of the lactone metabolite present in the sample, unless there is a chromatographic separation between the two compounds. This is true even if Q1 is operated under a unit-mass resolution. The implication of this type of interference, arising from the presence of both the [M + H](+) and [M + NH(4)](+) ions of a drug and its metabolite, to the selection of LC and MS conditions (including mass resolution) will be discussed using the data obtained with a model lactone drug and its hydroxy acid metabolite. PMID- 11006584 TI - Pyrolysis-gas chromatography/mass spectrometry of a coal extract and its fractions separated by planar chromatography: correlation of structural features with molecular mass AB - The structural characterisation of a coal liquefaction extract and its three fractions separated by planar chromatography has been described. Size exclusion chromatography showed the molecular mass distributions to become progressively larger with decreasing mobility on the plate. UV-fluorescence spectroscopy of the fractions indicated parallel increases in the sizes of polynuclear aromatic ring systems. Analysis by probe-mass spectrometry of the 'whole' coal extract showed the expected array of small polynuclear aromatic groups extending to m/z 450. The probe mass spectra of the lightest fraction ('mobile in pyridine and acetonitrile') showed similar features, except for effects due to vacuum drying to remove solvent. In sharp contrast, the two heaviest fractions ('mobile in pyridine and immobile in acetonitrile' and 'immobile in pyridine') showed no significant ions other than those from residual NMP solvent (m/z 98 and 99). Pyrolysis-gas chromatography/mass spectrometry of these two heaviest fractions showed only traces of aromatic compounds or fragments. The aromatic pyrolysis products of these fractions were too large and involatile to pass through the GC column. The major components observed in the pyrolysis-gas chromatography/mass spectrometry of the two heavy fractions were alkanes and alkenes, ranging between C10-C25. Since none of the samples contained free alkanes, alkenes or cycloalkanes before pyrolysis, they were generated during the pyrolysis step. The shifts of UV-fluorescence spectral intensity to shorter wavelengths with decreasing size indicated by size exclusion chromatography (SEC) provide direct evidence of differences in structure with changing molecular mass. This evidence strongly suggests that species identified as being of large molecular mass in this extract sample are not composed of molecular aggregates. It remains difficult to establish whether and when it would be legitimate to invoke molecular aggregates to explain the large molecular masses (MMs) identified here and in other work. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006585 TI - Mass spectra of new substituted 2-amino-4H-pyrans: a retro-Diels-Alder reaction pattern. AB - New substituted 2-amino-3-cyano-4H-pyrans have been studied by electron ionization (EI), chemical ionization (CI) and electrospray ionization (ESI) mass spectrometry. The retro-Diels-Alder reaction (RDA) is the main fragmentation pattern observed in the EI spectra forming an unsaturated ketone as the diene fragment. In contrast, a different RDA reaction takes place yielding an unsaturated amide as diene fragment together with the unsaturated ketone in the CI spectra. The MS/MS spectra obtained using an ESI source reveal that the favoured fragmentation by collision induced dissociation (CID) is the elimination of the substituent at the C4 position with formation of a stable pyrilium cation. PMID- 11006586 TI - Evaluation of automated single mass spectrometry to tandem mass spectrometry function switching for comprehensive drug profiling analysis using a quadrupole time-of-flight mass spectrometer. AB - A liquid chromatographic mass spectrometric strategy for systematic toxicological analysis (STA) is presented using the automatic 'on-the-fly' single mass spectrometry mode to tandem mass spectrometry mode (MS to MS/MS) switching abilities of a quadrupole time-of-flight (Q-TOF) instrument. During the chromatographic run, the quadrupole is initially set to transmit all masses until (an) ion(s) reaches a certain set threshold. Thereupon, the quadrupole automatically switches to the MS/MS mode, selecting the ion(s), which are subsequently fragmented in the high-efficiency hexapole collision cell, thus generating product ions that are further mass analyzed by the TOF. By limiting the TOF spectral accumulation time in the MS/MS mode to a statistically acceptable minimum, the quadrupole almost instantly switches back to the MS mode. Qualitative information, comprising the complementary MS ([M + H](+) ion mass) and MS/MS (informative product ion profile) data, as well as quantitative information obtained by integration of the MS extracted ion chromatogram(s), can be obtained in one single acquisition. Optimization of the automatic switching parameters, such as threshold, TOF spectral accumulation time, detection window and collision energy, was carried out by injection of a mix of 17 common drugs which were not necessarily baseline separated in the chromatographic system used. Indeed, the complete separation of the drugs is not deemed necessary since up to 8 different ions can 'simultaneously' be selected for MS/MS if they reach the preset criteria. In addition, the quantitative performance of the method was defined. In a second phase, the developed method was field-tested. To that end, the resulting data from extracts of urine samples were compared with and found to be in close concordance with those obtained by a standard toxicological analysis. This innovative approach clearly holds the potential for a substantial advance in the introduction of LC/MS in STA. PMID- 11006587 TI - Electron capture dissociation of singly and multiply phosphorylated peptides. AB - Analysis of phosphotyrosine and phosphoserine containing peptides by nano electrospray Fourier transform ion cyclotron resonance (FTICR) mass spectrometry established electron capture dissociation (ECD) as a viable method for phosphopeptide sequencing. In general, ECD spectra of synthetic and native phosphopeptides appeared less complex than conventional collision activated dissociation (CAD) mass spectra of these species. ECD of multiply protonated phosphopeptide ions generated mainly c- and z(.)-type peptide fragment ion series. No loss of water, phosphate groups or phosphoric acid from intact phosphopeptide ions nor from the c and z(.) fragment ion products was observed in the ECD spectra. ECD enabled complete or near-complete amino acid sequencing of phosphopeptides for the assignment of up to four phosphorylation sites in peptides in the mass range 1400 to 3500 Da. Nano-scale Fe(III)-affinity chromatography combined with nano-electrospray FTMS/ECD facilitated phosphopeptide analysis and amino acid sequencing from crude proteolytic peptide mixtures. PMID- 11006588 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of oligosaccharides derivatized by reductive amination and N,N-dimethylation. AB - Oligosaccharides were derivatized by reductive amination with benzylamine followed by N,N-dimethylation with methyl iodide and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and MALDI post-source decay (PSD) TOFMS. The resulting derivatives have a positive charge localized to the modified reducing end. The derivatization methodology was tested on maltoheptaose and three different human milk oligosaccharides. The approximate detection limit for the resulting carbohydrate derivatives was determined to be 50 fmol of the derivative loaded onto the target, corresponding to a tenfold increase in sensitivity compared with underivatized oligosaccharides. When the derivatives were analyzed by MALDI-PSD TOFMS the observed fragmentation pattern was dominated by fragment ions retaining the modified reducing terminus, thus simplifying the interpretation of the mass spectral data. PMID- 11006589 TI - Characterization of mesogen-jacketed liquid crystalline polymers by matrix assisted laser desorption/ionization time-of-flight mass spectrometry AB - For synthetic polymers, a proper sample preparation method is essential for successful characterization by matrix-assisted laser desorption/ionization time of-flight (MALDI-TOF) mass spectrometry. In this work, six synthetic mesogen jacketed liquid crystalline polymers (MJLCPs) with different main-chain, spacer and mesogenic units were investigated by MALDI-TOF mass spectrometry. Several factors that affect the analysis of these polymers were examined. These factors include matrices used, cationization salts used, the concentration of polymers, and the ratio of sample to matrix. After testing different conditions, we found a suitable sample preparation method for these six polymers. The number average molecular weight (M(n)), weight average molecular weight (M(w)) and polydispersity (PD) were calculated using data obtained in the linear mode. The end groups of the polymers were proposed using data obtained in reflectron mode. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006590 TI - Electrospray ionization mass spectra of amino acid phosphoramidates of adenosine. AB - Amino acid phosphoramidates of adenosine were synthesized and determined by positive and negative ion electrospray ionization mass spectrometry (ESI-MS) in conjunction with tandem mass spectrometry (MS/MS). The fragmentation pathways were investigated. In the positive ion mass spectra abundant characteristic fragment ions appeared, and many complementary ions were found. In the negative ion mass spectra only a few fragment ions were observed, and most of them contained phosphoryl groups. The results show that ESI-MS is a useful tool for structural determination of amino acid phosphoramidates of nucleosides. PMID- 11006591 TI - Application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to insoluble glucose oligomers in decomposed cellulose. AB - An analytical approach is described for the molecular weight characterization of glucose oligomers from cellulose which had been decomposed by hot-compressed water (HCW). Microcrystalline cellulose was decomposed to 18% of water insoluble fraction (WI) plus 82% of water soluble fraction (WS) by HCW (295 degrees C, 10 mL/min). The glucose oligomers in the WI were analyzed by pericullar anion exchange chromatography, and oligomers longer than pentamers were detected. These results were also confirmed by matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOFMS). Many peaks derived from glucose oligomers with molecular weights less than 4000 Da were detected in the spectra. Mass differences between peaks were 162 Da, as expected for polymers composed of glucose units. This appears to be the first report concerning molecular weight determination of water or organic solvent insoluble compounds like glucose oligomers from cellulose using MALDI-TOFMS. PMID- 11006592 TI - Advantages of using nested collision induced dissociation/post-source decay with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: sequencing of novel peptides from wasp venom. AB - We have examined the applicability of the 'nested' collision induced dissociation/post-source decay (CID/PSD) method to the sequencing of novel peptides from solitary wasps which have neurotoxic venom for paralyzing other insects. The CID/PSD spectrum of a ladder peptide derived from an exopeptidase digest was compared with that of the intact peptide. The mass peaks observed only in the CID/PSD spectrum of a ladder peptide were extracted as C-terminal fragment ions. Assignment of C-terminal fragment ions enabled calculation of N-terminal fragment masses, leading to differentiation between N-terminal fragment ions and internal fragment ions. This methodology allowed rapid and sensitive identification by removing ambiguity in the assignment of the fragment ions, and proved useful for sequencing unknown peptides, in particular those available as natural products with a limited supply. PMID- 11006593 TI - Measurement of 19-nortestosterone and its esters in equine plasma by high performance liquid chromatography with tandem mass spectrometry. AB - A high-performance liquid chromatographic-tandem mass spectrometric (HPLC/MS/MS) method for the determination of 19-nortestosterone and its esters (cyclopentanepropionate, phenylpropionate, and decanoate) in equine plasma is achieved using an atmospheric pressure chemical ionization (APCI) interface in selected reaction monitoring (SRM) mode. The two internal standards used were 16,16, 17-(2)H(3)-19-nortestosterone for 19-nortestosterone and methenolone acetate for its esters. The steroids studied were extracted from plasma samples with a mixture of diethyl ether/n-hexane (9:1, v/v). The quantification limits for 19-nortestosterone, 19-nortestosterone cyclopentanepropionate, 19 nortestosterone phenylpropionate, and 19-nortestosterone decanoate were 0.16, 5.0, 0.1, and 2.0 ng/mL, respectively, when 2 mL of plasma were used. The recoveries of most of the steroids were 71.6-101.0% except for the decanoate, which could be recovered to about 39.8%. The responses were linear, with correlation coefficients varying from 0.9897 to 0.9999 in the concentration range of 0.1 to 50.0 ng/mL for the steroids studied. When applied to equine (mare) plasma samples, the present method allowed detection of 19-nortestosterone up to 23 days after an intra-muscular injection of 400 mg as the decanoate. PMID- 11006594 TI - Effect of sample preparation methods on the analysis of dispersed polysaccharides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Effect of sample preparation methods on the matrix-assisted laser desorption/ionization (MALDI) analysis of dispersed polysaccharides is reported. By using the conventional drop-drying method, the measured number-averaged molecular weights were found to differ significantly from the values obtained from gel-permeation chromatography (GPC). These discrepancies were found to increase as the average masses of the polysaccharides increased. To understand the impact of the sample preparation method on the MALDI measurement, a dispersed dextran sample was separated into ten narrow-distribution fractions. Mixtures of different mass fractions were prepared and analyzed by using different mixing and preparation methods. By using the ratio of the signal intensity for the low-mass fraction to that of the high-mass fraction as an indicator, the impact of sample preparation conditions on the enhancement of low-mass components (or suppression of high-mass components) was determined. From the results obtained, it is postulated that the difference in solubilities between the low-mass and high-mass components for dispersed polymers might be large enough to induce an enhancement of low-mass components (or suppression of high-mass components) at the surface of the crystals during sample crystallization in the drop-drying method. PMID- 11006595 TI - A liquid chromatography tandem mass spectrometric method for in vivo dose monitoring of diepoxybutane, a metabolite of butadiene. AB - 1,3-Butadiene, a common air pollutant formed in the combustion of organic matter, has been assessed by the U.S. EPA to be a strongly carcinogenic compound. This risk assessment is very uncertain because of the lack of information on the dose of the powerful carcinogenic metabolite diepoxybutane (DEB). This report presents an analytical method for in vivo dose monitoring of a unique marker for DEB. For a large number of alkylating agents in vivo doses are monitored by measurement by gas chromatography/mass spectrometry (GC/MS) of adducts to N-terminal valine in hemoglobin (Hb), using a modified Edman degradation method. This method is applicable to monofunctional epoxides from butadiene. However, in reaction with N terminal valine, DEB forms an adduct which is ring-closed to a pyrrolidine, N,N (2,3-dihydroxy-1,4-butadiyl)valine, with a tertiary amino group that prevents detachment of the alkylated valine by the Edman reagent. Therefore a method has been developed based on the analysis by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) of the N-modified N-terminal peptides enriched after trypsin digestion of globin. In this study Hb samples from mice injected intraperitoneally with (+/-)-DEB were examined qualitatively and quantitatively with regard to the ring-closed adduct. The N-terminal pyrrolidine heptapeptide was identified in treated mice. The highest adduct levels were obtained in samples from animals given the highest dose of DEB and the adduct levels were below the detection level in control mice. PMID- 11006596 TI - Using a superconducting tunnel junction detector to measure the secondary electron emission efficiency for a microchannel plate detector bombarded by large molecular ions AB - An energy-sensitive superconducting tunnel junction (STJ) detector was used to measure the secondary electron emission efficiency, epsilon(e), for a microchannel plate (MCP) detector bombarded by large (up to 66 kDa), slow moving (<40 km/s) molecular ions. The method used is new and provides a more direct procedure for measuring the efficiency of secondary electron emission from a surface. Both detectors were exposed simultaneously to nearly identical ion fluxes. By exposing only a small area of the MCP detector to ions, such that the area exposed was effectively the same as the size of the STJ detector, the number of ions detected with each detector were directly comparable. The STJ detector is 100% efficient for detecting ions in the energy regime investigated and therefore it can be used to measure the detection efficiency and secondary electron emission efficiency of the MCP. The results are consistent with measurements made by other groups and provide further characterization of the loss in sensitivity noted previously when MCP detectors have been used to detect high-mass ions. Individual molecular ions of mass 66 kDa with 30 keV kinetic energy were measured to have about a 5% probability of producing one or more electrons when impacting the MCP. When ion energy was reduced to 10 keV, the detection probability decreased to 1 %. The secondary electron yield was calculated from the secondary electron emission efficiency and found to scale linearly with the mass of the impinging molecular ion and to about the fourth power of ion velocity. Secondary electrons were observed for primary ion impacts >5 km/s, regardless of mass, and no evidence of a velocity (detection) threshold was observed. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006597 TI - Effects of ultraviolet irradiation on melanogenesis from tyrosine, Dopa and dopamine: a matrix-assisted laser desorption/ionization mass spectrometric study. AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry experiments were applied to study the influence of ultraviolet (UV) irradiation in melanogenesis. Samples were prepared starting from three different precursors, tyrosine, Dopa and dopamine, in the presence or absence of tyrosinase, the enzyme responsible for the synthesis of melanin. Enzymatic reactions were carried out for 10, 30, 60 and 120 min under UV irradiation at 365 nm, and aliquots were then immediately ultrafiltered and lyophilized. Samples obtained by irradiation of tyrosine solution revealed the formation of 5,6-dihydroxyindole (DHI) oligomers up to pentamers at 120 min; the reaction kinetics were markedly enhanced in the presence of tyrosinase. In the case of Dopa, UV irradiation favored melanogenesis only in the presence of the enzyme; in this case, many reaction pathways were activated, originating various oligomeric species of Dopa, DHI and 5,6 dihydroxyindole-2-carboxylic acid (DHICA). Conversely, when dopamine was used as tyrosinase substrate under UV light, mechanisms of melanogenesis different from those generated by simple enzymatic reaction without irradiation were not activated, as the same oligomeric species were present. PMID- 11006598 TI - Characterization of zirconium complexes of interest in spent nuclear fuel reprocessing by electrospray ionization mass spectrometry AB - Liquid-liquid extraction of zirconium, one of the most important fission products, was followed using electrospray ionization mass spectrometry under conditions simulating reprocessing of nuclear spent fuel. Zr(IV) can precipitate from the organic phase after extraction by dibutylphosphoric acid (HDBP), the most common degradation product of tributylphosphate (TBP) radiolysis. Different complexes were detected with electrospray used in positive or negative ion modes, according to the extraction conditions such as the ligand/metal ratio. Stoichiometry of the Zr(IV) complexes was determined by combining isotopic labeling [H(15)NO(3)] of the aqueous phase in the extraction system and tandem mass spectrometry experiments. These results were compared with the species observed using other techniques reported in the literature. The mechanisms of ionization/desorption of these complexes are proposed depending on the organic ligand character (neutral (L) such as TBP, or acidic (HL') such as HDBP), and the ionization mode used. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006599 TI - Oxygen attachment to metal complex ions during their collision induced dissociation in ion trap PMID- 11006600 TI - Editorial PMID- 11006601 TI - Tandem mass spectrometry: dissociation of ions by collisional activation AB - This review presents a brief historical introduction to the development of tandem mass spectrometry and its principal applications. It is placed in the context of the general principles underlying mass spectrometry, particularly the relationships between internal energy and fragmentation kinetics. The center-of mass framework is presented as a convenient means of applying conservation of momentum to the energy transfer problem in tandem mass spectrometry as a means of deducing energy transfer in the collisional activation step and kinetic energy release as activated ions dissociate into fragment ions and neutrals. The principles of molecular beam methods are summarized and illustrative examples are given for which definitive information on reaction dynamics is available. The importance of scattering-very little appreciated in early discussions of tandem mass spectrometry-is shown to be the natural consequence of impulsive collisions, which appears to be a general mechanism for energy exchange in collisional activation. It is shown that the average energy transferred in single collisions is much less than the theoretical maximum given by the center-of-mass collision energy and the Massey criterion is presented as a simplistic rationale for understanding the essentially exponential decline in the energy transfer function above and below the relative velocity at which the probability for energy transfer is maximized. The issues of energy transfer in collisions of large molecular ions with low-mass neutrals are reviewed and a general description of energy transfer in multiple collisions is presented. It is shown that the center of-mass and Massey criterion limitations are pragmatically overcome by multiple collision activation in ion traps. Surface-induced dissociation is presented as a viable alternative to multiple collision activation which is especially attractive for activation of large molecular ions. Finally, a few of the emerging dynamics principles governing energy transfer and dissociation of peptides are summarized. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006602 TI - Determination of phenothiazines in human body fluids by solid-phase microextraction and liquid chromatography/tandem mass spectrometry. AB - Eleven phenothiazine derivatives with heavy side-chains were found to be extractable from human whole blood and urine samples by solid-phase microextraction (SPME) with a polyacrylate-coated fiber. The fiber was then injected into the desorption chamber of an SPME-liquid chromatography (LC) interface for LC/tandem mass spectrometry (MS/MS) with positive ion electrospray (ES) ionization. All compounds formed base peaks due to [M + 1](+) ions by LC/ES MS/MS. By use of LC/ES-MS/MS, the product ions produced from each [M + 1](+) ion showed base peaks due to side-chain liberation. Selected reaction monitoring (SRM) and selected ion monitoring (SIM) were compared for the detection of the 11 phenothiazine derivatives from human whole blood and urine. SRM showed much higher sensitivity than SIM for both types of sample. Therefore, a detailed procedure for the detection of drugs by SRM with SPME-LC/MS/MS was established and carefully validated. The extraction efficiencies of the 11 phenothiazine derivatives spiked into whole blood and urine were 0. 0002-0.12 and 2.6-39.8%, respectively. The regression equations for the 11 phenothiazine derivatives showed excellent linearity with detection limits of 0.2-200 ng ml(-1) for whole blood and 4-22 pg ml(-1) for urine. The intra- and inter-day precisions for whole blood and urine samples were not greater than 15.1%. The data obtained after oral administration of perazine or flupentixol to a male subject are presented. PMID- 11006603 TI - Negative ion chemical ionization for the determination of methylphenidate in human plasma by stable isotope dilution gas chromatography/mass spectrometry. AB - A sensitive and specific method for the determination of methylphenidate in human plasma is presented. Methylphenidate was extracted from plasma by solvent extraction with hexane at pH 9.3 and derivatized to its heptafluorobutyrate derivative. The derivative was measured by gas chromatography/negative ion chemical ionization mass spectrometry without any further purification. Using this detection mode, a diagnostically useful fragment ion at m/z 369 was obtained at high relative abundance. (18)O(2)-labelled methylphenidate was used as an internal standard and its rapid and facile preparation from the unlabeled compound is described. Calibration graphs were linear within the range 0.14-18.25 ng ml(-1). The inter-assay precision was 8.7% (0.14 ng ml(-1)) and 3.1% (4.56 ng ml(-1)) and the intra-assay variability was 1.3% (0.14 ng ml(-1)) and 0.4% (4.56 ng ml(-1)). Accuracy determinations showed deviations of +0.7% (0.14 ng ml(-1)) and -2.5% (4.56 ng ml(-1)). The method is rugged, rapid and robust and has been applied to the batch analysis of methylphenidate during pharmacokinetic profiling of the drug. PMID- 11006604 TI - Comparison of a jet separator and an open splitter as an interface between a multi-capillary gas chromatographic column and a time-of-flight mass spectrometer AB - A gas chromatographic/time-of-flight mass spectrometric (GC/TOFMS) interface is being developed for fast on-line analysis utilizing multi-capillary column technology. A variable gap-distance jet separator has been constructed and its performance compared with that of a commercially supplied post-column open splitter recommended for use between the multi-capillary column and a mass spectrometer. Both interfaces were found to be compatible with the GC/TOFMS system at high carrier gas flow-rates, facilitating high-speed and high resolution separations. The systems were investigated and tested with a mixture of volatile organic compounds (VOCs) with molecular masses from 85 to 166: dichloromethane, toluene, m-dichlorobenzene, o-dichlorobenzene and tetrachloroethylene. The optimum tip-to-tip gap distance corresponding to the highest efficiency of the jet separator was found to be 0.030 mm for each compound at carrier gas flow-rates of 20, 40 and 60 ml min(-1) giving, in the ion source housing, ion gauge pressure readings of 1.6 x 10(-6), 5.0 x 10(-6) and 5.8 x 10(-6) mbar, respectively. The efficiency of the jet separator (10-30% yields) was significantly higher than that of the open splitter (6-9% yields). The observation that the open splitter did not provide a constant flow-rate to the ion source was not in agreement with the manufacturer's specifications. A method for measuring the gas flow-rates in all parts of the equipment is described. The correlation between yield in the jet separator and molecular mass for the heterogeneous set of compounds studied was found to be less linear than usually reported for homologous series of compounds in jet separator studies. The result suggests that the pressure conditions in the jet may be sufficient for the separation process to be partly controlled by diffusion rather than predominately by effusion. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006606 TI - Characterization of isopoly metal oxyanions using electrospray time-of-flight mass spectrometry AB - Electrospray time-of-flight mass spectrometry was used to study dilute (10(-3) M) solutions of tungstate, vanadate and perrhenate oxyanions under acidic and basic conditions. The electrospray mass spectra of tungstate and vanadate solutions indicate the presence of protonated monomeric oxyanions in basic solution (pH approximately 10) and protonated polymeric species in acidic solutions (pH approximately 4). Monomeric perrhenate was the only species detected in both acidic and basic perrhenate solutions. For all three systems, the core metalate species detected by electrospray were consistent with reported speciation in aqueous solutions. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11006605 TI - Isolation from rat urine and human liver microsomes and identification by electrospray and nanospray tandem mass spectrometry of new malagashanine metabolites. AB - Malagashanine has been isolated from indigenous madagascan Strychnos myrtoides alkaloids used traditionally to treat malaria. This alkaloid was found to enhance the action of chloroquine against chloroquine-resistant strains of Plasmodium falciparum when combined with classical antimalarial drugs (chloroquine, quinine). The present study was carried out in order to investigate by electrospray mass and tandem mass spectrometry and NMR spectroscopy the structure of two new metabolites isolated from rat urine and human liver microsomes. We were able to demonstrate the presence of two new metabolites of malagashanine corresponding to a malagashanine N-demethylated metabolite and to the oxidation of malagashanine in the alpha-position of the N-methyl group to produce a carbinolamine function. The latter metabolite may be subject to ring and open chain tautomerism effects and dimeric species were detected in the electrospray mass spectrum. PMID- 11006607 TI - Use of an automated chromium reduction system for hydrogen isotope ratio analysis of physiological fluids applied to doubly labeled water analysis. AB - The doubly labeled water method is commonly used to measure total energy expenditure in free-living subjects. The method, however, requires accurate and precise deuterium abundance determinations, which can be laborious. The aim of this study was to evaluate a fully automated, high-throughput, chromium reduction technique for the measurement of deuterium abundances in physiological fluids. The chromium technique was compared with an off-line zinc bomb reduction technique and also subjected to test-retest analysis. Analysis of international water standards demonstrated that the chromium technique was accurate and had a within-day precision of <1 per thousand. Addition of organic matter to water samples demonstrated that the technique was sensitive to interference at levels between 2 and 5 g l(-1). Physiological samples could be analyzed without this interference, plasma by 10000 Da exclusion filtration, saliva by sedimentation and urine by decolorizing with carbon black. Chromium reduction of urine specimens from doubly labeled water studies indicated no bias relative to zinc reduction with a mean difference in calculated energy expenditure of -0.2 +/- 3.9%. Blinded reanalysis of urine specimens from a second doubly labeled water study demonstrated a test-retest coefficient of variation of 4%. The chromium reduction method was found to be a rapid, accurate and precise method for the analysis of urine specimens from doubly labeled water. PMID- 11006609 TI - Collision-induced dissociation spectra versus collision energy (collision-induced dissociation curve) using a quadrupole ion trap mass spectrometer. II.(1) loss of CO from ionized o-, m- and p-anisoyl fluoride, CH(3)OC(6)H(4)COF(+.) PMID- 11006608 TI - Quantitation of simvastatin and its beta-hydroxy acid in human plasma by liquid liquid cartridge extraction and liquid chromatography/tandem mass spectrometry. AB - A sensitive and reliable procedure for the simultaneous determination of simvastatin (SV) and its active beta-hydroxy acid metabolite (SVA) in human plasma was developed and validated. The analytes were extracted simultaneously from 0.5 ml aliquots of human plasma samples by methyl tert-butyl ether (MTBE) via Chem Elut cartridge extraction [also called liquid-solid extraction (LSE) or liquid-liquid cartridge extraction (LLCE)], separated through a Kromasil C(18) column (50 x 2 mm i.d. 5 microm) and detected by tandem mass spectrometry with a turbo ionspray interface. Stable isotope-labeled SV and SVA, (13)CD(3)-SV and (13)CD(3)-SVA, were used as internal standards. SV and SVA were detected in positive and negative ion modes, respectively, via within-run polarity switching. The use of Chem Elut cartridges not only provided a simple and efficient means of plasma sample extraction but also successfully reduced the interconversion between SV and SVA to an undetectable (for lactonization of SVA) or negligible (<0.07%, for hydrolysis of SV) level. The method showed excellent reproducibility, with intra- and inter-assay precisions <4.5% (RSD), and intra- and inter-assay accuracy between 94% and 107% of nominal values, for both analytes. The extraction recoveries were 78% and 87% on average for SV and SVA, respectively. The analyte was found to be stable in plasma through three freeze ( 70 degrees C)-thaw (4 degrees C) cycles and for at least 3 h under bench-top storage condition in an ice-bath (4 degrees C), and also in the reconstitution solution at 4 degrees C for at least 24 h. The method has a lower limit of quantitation (LOQ) of 50 pg ml(-1) with a linear calibration range of 0.05-50 ng ml(-1) for both analytes, and has proved to be very reliable for the analysis of clinical samples. PMID- 11006610 TI - Current literature in mass spectrometry. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of mass spectrometry. Each bibliography is divided into 11 sections: 1 Books, Reviews & Symposia; 2 Instrumental Techniques & Methods; 3 Gas Phase Ion Chemistry; 4 Biology/Biochemistry: Amino Acids, Peptides & Proteins; Carbohydrates; Lipids; Nucleic Acids; 5 Pharmacology/Toxicology; 6 Natural Products; 7 Analysis of Organic Compounds; 8 Analysis of Inorganics/Organometallics; 9 Surface Analysis; 10 Environmental Analysis; 11 Elemental Analysis. Within each section, articles are listed in alphabetical order with respect to author (3 Weeks journals - Search completed at 28th. June 2000) PMID- 11006611 TI - Prion diseases. PMID- 11006612 TI - Future employment of allopathic medical graduates. PMID- 11006613 TI - Doctor-population ratios. PMID- 11006614 TI - Thalassaemia control in developing countries--the Sri Lankan perspective. AB - Beta thalassaemia is a Mendelian recessive disorder. The economic and social cost of the disease is high due to patients' life long need for monthly blood transfusions and treatment with desferrioxamine, an iron chelating agent. Wider availability of cheaper drugs is on the horizon. If there is no concomitant reduction in the number of new thalassaemia major births, there will be a cumulative increase in numbers requiring treatment. The frequency and severity, and the economic and social costs of thalassaemia, support the case for the introduction of a carrier screening and counselling program in Sri Lanka. A three pronged plan emphasising professional, political and public education in outlined. PMID- 11006615 TI - Suicide in Sri Lanka: points to ponder. PMID- 11006616 TI - Suicide: a Socratic revenge. AB - 2400 years have passed since the occurrence in Athens, Greece of one of the famous suicides recorded in human history. This autobiographical essay provides a montage on the history of suicide, with snippets from the final hours of Socrates, as described by Plato. Suicide in contemporary Japanese culture is also explored briefly, with reference to the deaths of internationally acclaimed movie directors Akira Kurosawa and Juzo Itami. The author also questions why no researcher has yet been honoured for the past 99 years with a Medicine Nobel prize for his or her work on suicidology or thanatology. PMID- 11006617 TI - Paradoxical progression of intracranial tuberculous lesions during treatment. PMID- 11006618 TI - A case of hereditary sensory and autonomic neuropathy (HSAN) type II. AB - We describe a case of hereditary sensory and autonomic neuropathy (HSAN) type II in a child with a penetrating foot ulcer, acral sensory impairment, and anhidrosis. This is the first documentation of HSAN in Sri Lanka. PMID- 11006619 TI - Mixed medullary follicular carcinoma--an unusual thyroid neoplasm. PMID- 11006620 TI - Acute primary axonal Guillain-Barre syndrome. PMID- 11006621 TI - Sleeping sickness and teaching of medical students. PMID- 11006622 TI - Sleeping sickness in Sri Lanka. PMID- 11006623 TI - Sleeping sickness in Sri Lanka. PMID- 11006624 TI - Inhaled steroids for childhood asthma. PMID- 11006625 TI - Metformin in diabetes mellitus: are we choosing the right patients? PMID- 11006626 TI - Heat stroke in young adults: in reply. PMID- 11006627 TI - A cautionary tale: intra-amniotic injection of methylene blue. PMID- 11006628 TI - Impact of new diagnostic criteria for diabetes on its prevalence in a hospital population. PMID- 11006629 TI - The Neurospora am gene and NADP-specific glutamate dehydrogenase: mutational sequence changes and functional effects--more mutants and a summary. AB - A further series of mutant am alleles, encoding potentially active NADP-specific glutamate dehydrogenase (GDH) and capable of complementation in heterocaryons, have been characterized with respect to both GDH properties and DNA sequence changes. Several mutants previously studied, and some of their same-site or second-site revertants, have also been sequenced for the first time. We present a summary of what is known of the properties of all am mutants that have been defined at the sequence level. PMID- 11006630 TI - Temperature-induced alteration of the polytene X chromosome structure in male larvae of the strain In(1)BM2 (reinverted) of Drosophila melanogaster. AB - In Drosophila melanogaster, the polytene X chromosome of male third instar larva appears twice as wide as an unpaired female X chromosome or an autosome. This characteristic morphology of the male X chromosome is correlated with the increased rate of transcription of the sex-linked genes, which ensures gene dosage compensation. In male third instar larvae of the strain In(1)BM2 (reinverted), polytene nuclei manifest unusually puffy X chromosomes at 18 +/- 1 degrees C. Such 'puffy X' chromosomes are pompons, that is, despite the increased width of the chromosome, transcription remains at the wild-type level. This characteristic is a caveat to the invariable correlation between polytene chromosome puffs and transcription, and suggests that the mutant X chromosomes arise due to perturbation of a pathway that controls the structure but not the transcription of the polytene X chromosome. In this report we present evidence that the pompons of In(1)BM2 (reinverted) arise due to spiralization of the male X chromosome, which results in condensing of the chromosome. This unusual structural alteration can be induced only in male larvae of this strain, at the third instar larval stage, through temperature shifts from 24 +/- 1 degrees C to 18 +/- 1 degrees C and during recovery from cold shock. Furthermore, extract from male adult, pupae and third instar larvae can induce chromosome condensation in wild-type larvae in vitro. This new evidence not only explains the absence of correlation between chromosome width and transcription of the pompons of In(1)BM2 (reinverted), but also suggests that the chromosomal rearrangement perturbs a pathway that regulates the condensation of chromosomes. PMID- 11006631 TI - Polymorphism of M factors in populations of the housefly, Musca domestica L., in Turkey. AB - M factors, which determine maleness in Musca domestica, were found on the second, third, fourth and fifth linkage groups in housefly populations of Turkey. As in European populations, the male-determining factor was more frequently located on linkage group III (MIII). Some males homozygous or double heterozygous for M factors were identified. Deviations from a 1:1 sex ratio in favour of males, as well as mosaics for somatic marker mutations and sexual mosaics (gynandromorphs), were also observed. The results reveal an extensive polymorphism in the sex determining system. PMID- 11006632 TI - Quantitative trait loci for directional but not fluctuating asymmetry of mandible characters in mice. AB - Non-directional variation in right minus left differences in bilateral characters, referred to as fluctuating asymmetry (FA), often has been assumed to be largely or entirely environmental in origin. FA increasingly has been used as a measure of developmental stability, and its presumed environmental origin has facilitated the comparisons of populations believed to differ in their levels of stability. Directional asymmetry (DA), in which one side is consistently larger than the other, has been assumed to be at least partially heritable. Both these assumptions were tested with interval mapping techniques designed to detect any quantitative trait loci (QTLs) affecting FA or DA in 15 bilateral mandible characters in house mice resulting from a cross of the F1 between CAST/Ei (wild strain) and M16i (selected for rapid growth rate) back to M16i. For purposes of the analysis, all mandibles were triply measured and 92 microsatellite markers were scored in a total of 350 mice. No significant QTLs were found for FA, but three QTLs significantly affected DA in several characters, confirming both assumptions. The QTLs for DA were similar in location to those affecting the size of several of the mandible characters, although they accounted for an average of only 1% of the total phenotypic variation in DA. PMID- 11006634 TI - The effect of subdivision on variation at multi-allelic loci under balancing selection. AB - Simulations are used to investigate the expected pattern of variation at loci under different forms of multi-allelic balancing selection in a finite island model of a subdivided population. The objective is to evaluate the effect of restricted migration among demes on the distribution of polymorphism at the selected loci at equilibrium, and to compare the results with those expected for a neutral locus. The results show that the expected number of alleles maintained, and numbers of nucleotide differences between alleles, are relatively insensitive to the migration rate, and differentiation remains low even under very restricted migration. However, nucleotide divergence between copies of functionally identical alleles increases sharply when migration decreases. These results are discussed in relation to published surveys of allelic diversity in MHC and plant self-incompatibility systems, and to the possibility of inferring ancient population genetic events and processes. In addition, it is shown that, for sporophytic self-incompatibility systems, it is not necessarily true in a subdivided population that recessive alleles are more frequent than dominant ones. PMID- 11006633 TI - Characterization and differential expression of a human gene family of olfactomedin-related proteins. AB - Olfactomedin-related proteins are secreted glycoproteins with conserved C terminal motifs. Olfactomedin was originally identified as the major component of the mucus layer that surrounds the chemosensory dendrites of olfactory neurons. Homologues were subsequently found also in other tissues, including the brain and in species ranging from Caenorhabditis elegans to Homo sapiens. Most importantly, the TIGR/myocilin protein, expressed in the eye and associated with the pathogenesis of glaucoma, is an olfactomedin-related protein. The prevalence of olfactomedin-related proteins among species and their identification in different tissues prompted us to investigate whether a gene family exists within a species, specifically Homo sapiens. A GenBank search indeed revealed an entire human gene family of olfactomedin-related proteins with at least five members, designated hOlfA through hOlfD and the TIGR/myocilin protein. hOlfA corresponds to the rat neuronal AMZ protein. Phylogenetic analyses of 18 olfactomedin-related sequences resolved four distinct subfamilies. Among the human proteins, hOlfA and hOlfC, both expressed in brain, are most closely related. Northern blot analyses of 16 human tissues demonstrated highly specific expression patterns: hOlfA is expressed in brain, hOlfB in pancreas and prostate, hOlfC in cerebellum, hOlfD in colon, small intestine and prostate and TIGR/myocilin in heart and skeletal muscle. The link between TIGR/myocilin and ocular hypertension and the expression of several of these proteins in mucus-lined tissues suggest that they play an important role in regulating physical properties of the extracellular environment. Future studies can now assess whether other members of this gene family, like TIGR/myocilin, are also associated with human disease processes. PMID- 11006635 TI - The effect of hitch-hiking on genes linked to a balanced polymorphism in a subdivided population. AB - The effect of multi-allelic balancing selection on nucleotide diversity at linked neutral sites was investigated by simulations of subdivided populations. The motivation is to understand the behaviour of self-recognition systems such as the MHC and plant self-incompatibility. For neutral sites, two types of subdivision are present: (1) into demes (connected by migration), and (2) into classes defined by different functional alleles at the selected locus (connected by recombination). Previous theoretical studies of each type of subdivision separately have shown that each increases diversity, and decreases the relative frequencies of low-frequency variants, at neutral sites or loci. We show here that the two types of subdivision act non-additively when sampling is at the whole population level, and that subdivision produces some non-intuitive results. For instance, in highly subdivided populations, genetic diversity at neutral sites may decrease with tighter linkage to a selected locus or site. Another conclusion is that, if there is population subdivision, balancing selection leads to decreased expected FST values for neutral sites linked to the selected locus. Finally, we show that the ability to detect balancing selection by its effects on linked variation, using tests such as Tajima's D, is reduced when genes in a subdivided population are sampled from the total population, rather than within demes. PMID- 11006636 TI - Effects of population structures and selection strategies on the purging of inbreeding depression due to deleterious mutations. AB - Stochastic simulations were run to compare the effects of nine breeding schemes, using full-sib mating, on the rate of purging of inbreeding depression due to mutations with equal deleterious effect on viability at unlinked loci in an outbred population. A number of full-sib mating lines were initiated from a large outbred population and maintained for 20 generations (if not extinct). Selection against deleterious mutations was allowed to occur within lines only, between lines or equal within and between lines, and surviving lines were either not crossed or crossed following every one or three generations of full-sib mating. The effectiveness of purging was indicated by the decreased number of lethal equivalents and the increased fitness of the purged population formed from crossing surviving lines after 20 generations under a given breeding scheme. The results show that the effectiveness of purging, the survival of the inbred lines and the inbreeding level attained are generally highest with between-line selection and lowest with within-line selection. Compared with no crossing, line crossing could lower the risk of extinction and the inbreeding coefficient of the purged population substantially with little loss of the effectiveness of purging. Compromising between the effectiveness of purging, and the risk of extinction and inbreeding coefficient, the breeding scheme with equal within- and between-line selection and crossing alternatively with full-sib mating is generally the most desirable scheme for purging deleterious mutations. Unless most deleterious mutations have relatively large effects on fitness in species with reproductive ability high enough to cope with the depressed fitness and thus increased risk of extinction with inbreeding, it is not justified to apply a breeding programme aimed at purging inbreeding depression by inbreeding and selection to a population of conservation concern. PMID- 11006637 TI - Fine-mapping of quantitative trait loci in half-sib families using current recombinations. AB - Two groups of methods are being developed to fine-map quantitative trait loci (QTLs): identity-by-descent methods or methods using historical recombinations, and genetic chromosome dissection methods or methods utilizing current recombinations. Here we propose two methods that fall into the second group: contrast mapping and substitution mapping. A QTL has previously been detected via linkage mapping in a half-sib design (granddaughter or daughter design), and sires (grandsires) likely to be heterozygous at the QTL have been identified. A sire (grandsire) and its recombinant offspring are then genotyped for a series of ordered markers spanning the initial marker interval. Offspring are grouped by paternal multi-marker haplotype with haplotypes differing in the location of the recombination event. In the contrast method, contrasts between the phenotypic averages of haplotypes or offspring groups are calculated which correspond to marker intervals within the original interval. The expected value of the contrast for the true QTL interval is always maximum, hence the interval with maximum observed contrast is assumed to contain the QTL. Alternative statistics for determining the interval most likely to contain a QTL are presented for contrast mapping, as well as a bootstrap estimation of the probability of having identified the correct interval. For an initial marker bracket of 20 cM and 10 additional equidistant markers, the probability of assigning the QTL to the correct 2 cM marker interval or to a combined 4 cM interval was calculated. For substitution effects of 0.093, 0.232, 0.464, 0.696 and 0.928 (in additive genetic SD), power values near 0.14, 0.26, 0.48, 0.67 and 0.80 (0.25, 0.53, 0.86, 0.97 and 0.99) are achieved for a family of 200 (1000) sons, respectively. In substitution mapping, QTL segregation status of recombinant sons must be determined using daughter genotyping. Combinations of two haplotypes with their segregation status are required to assign the QTL to an interval. Probabilities of correct QTL assignment were calculated assuming absence of the mutant QTL allele in dams of sons. For a 2 cM interval and a QTL at the midpoint of an interval, power near 0.95 (0.90) is reached when the number of recombinant sons is 70 (60), or total number of sons is 424 (363). For QTL positions away from the midpoint, power decreases but can be improved by combining marker intervals. For a QTL located halfway to the midpoint, and 182 sons in a family resulting in 30 recombinant sons, probability is 0.94 for assignment to either a 2 cM or a combined 4 cM interval. Effect of type I and type II errors in segregation status determination on power of QTL assignment was found to be small. Errors in segregation status due to QTL segregation in dams have an impact if the frequency of the mutant QTL allele is intermediate to high. PMID- 11006638 TI - Mapping quantitative trait loci in tetraploid populations. AB - Knowledge of quantitative trait locus (QTL) mapping in polyploids is almost void, albeit many exquisite strategies of QTL mapping have been proposed and extensive investigations have been carried out in diploid animals and plants. In this paper we develop a simple algorithm which uses an iteratively reweighted least square method to map QTLs in tetraploid populations. The method uses information from all markers in a linkage group to infer the probability distribution of QTL genotype under the assumption of random chromosome segregation. Unlike QTL mapping in diploid species, here we estimate and test the compound 'gametic effect', which consists of the composite 'genic effect' of alleles and higher order gene interactions. The validity and efficiency of the proposed method are investigated through simulation studies. Results show that the method can successfully locate QTLs and separates different sources (e.g. additive and dominance) of variance components contributed by the QTLs. PMID- 11006639 TI - High copy numbers of multiple transposable element families in an Australian population of Drosophila simulans. AB - Sudden mobilization of transposable elements in Drosophila is a well-reported phenomenon but one that usually affects no more than a few elements (one to four). We report here the existence of a D. simulans natural population (Canberra) from Australia, which had high copy numbers for various transposable elements (transposons, LTR retrotransposons and non-LTR retrotransposons). The impact of transposable elements on the host genome and populations is discussed. PMID- 11006640 TI - [Frequency of colicin and hemolysins in Escherichia coli isolated from pregnant patients with urinary tract infection, symptomatic and asymptomatic]. AB - Colicin production was studied among 137 Escherichia coli strains isolated from pregnant patients with symptomatic or asymptomatic, urinary tract infection (IVU). The observed colicinogeny frequency was 72.92% (n = 96), among the symptomatic patients and 29.26% (n = 41) among the asymptomatic group. The most frequently identified colicins from symptomatic patients were: V (23.9%), A (18.7%) and E1 (17.7%) and within the patients with asymptomatic IVU the most frequently observed colicins were A (9.7%), E1 (17.0%) y V (2.4%). The major frequency of colicin V among the E. coli strains isolated from the symptomatic group against the asymptomatic one, was statistical significant (p > or = 0.025). The results on the observed frequencies of colicins E1 and A were not statistical significant. Among the 137 studied E. coli strains, 37.2% were hemolytic. 83.3% of the colicin V producing strains (n = 24) were hemolytic, among the strains producing other colicin different than colicin V 34% (n = 58) were hemolytic and 12.0% of non colicinogenic strains were hemolytic. These results were statistical significant (p > 0.05). The present data, suggest that colicins production is an important pathogenicity factor among the E. coli strains, specially for those strains producing colicin V. The observed association among hemolysin and colicin V production may be an interesting pathogenicity factor which suggests an increasing ability of uropathogenic strains to produce symptomatic urinary tract infections. PMID- 11006641 TI - [Non-immunologic hydrops. Report of 2 cases]. AB - Non immune hydrops is an uncommon entity, it is observed in one per 2,000 to 3,500 alive birth. It can be idiopathic or secondary to different pathologies, the most important examples are: congenital malformations, cromosompathies or viral infections. OBJECTIVE: To show our therapeutic and diagnostic experience in two cases of non immune hydrops. STUDY DESIGN: To present a case of spontaneous reversion of non immune hydrops secondary to parvovirus B 19 infection and other case secondary to congenital cardiopathy. RESULTS: We found spontaneous reversion of a case of non immune hydrops due to a parvovirus B 19 infection and infant was normal but the infant with congenital cardiopathy died ten minutes after birth. CONCLUSIONS: Two of the most important causes of non immune hydrops are cardiopathies and infections, specially parvovirus which is responsible of poor perinatal outcome but there is a remote possibility of spontaneous reversion because it is important to make a specific diagnose and to make the expected management. PMID- 11006642 TI - [Methods of pedicle hemostasis in laparoscopically-assisted vaginal hysterectomy (VAVH). Evaluation of results]. AB - The objective was to evaluate whether use of different techniques of hemostasia have effect on laparoscopically assisted vaginal hysterectomy (LAVH) morbidity. Was performed a review of 53 LAVH cases, parameters analyzed were type of hemostasia method and surgical complications. There were five patients with transoperative bleeding and two with incidental bladder lesion, that were managed successfully without complications. There were no significant differences between types of hemostasia with respect to transoperative bleeding. There were no mayor surgical complications in the studied patients. As conclusion LAVH allows patients a expedite recovery with low postoperative complications. All the methods used in this work to perform hemostasia in LAVH are safe and effective for patients with habitual hysterectomy indications. PMID- 11006643 TI - [Prenatal corticosteroids. Use and attitudes of the gynecology-obstetrics medical staff]. AB - OBJECTIVES: 1. To evaluate the frequency of use of antenatal corticosteroids (AC) and the reasons for withholding them. 2. To estimate the knowledge and attitudes, regarding AC, from obstetricians, in a large Women and Babies Hospital. METHODS: Through a prospective, observational and transversal study we evaluated: 1. The frequency of use of AC in pregnant with preterm labor (PL) and in mothers of preterm infants (P1). When AC were not given, the reasons were recorded. 2. With an anonymous inquiry we assessed the knowledge and attitude of obstetricians regarding AC, exploring different concepts such as usefulness, frequency of use, indications, risks and contraindications. Descriptive statistics was used. RESULTS: We studied 364 patients (240 mothers of Pl and 124 pregnant women). The mean gestational age (GA) was 28.9 + 2.4 weeks. The AC were used only in 136 out of 364 (37.4%) women, and diminished to 30.9% (102/330), if are excluded 34 patients who did not receive AC because imminent delivery. Another reasons for not using AC were irregular uterine activity (18.9%), prolonged rupture of membranes without chorioamnioitis (11.4%), and GA lower than 27 weeks (11.4%) and toxemia (9.6%). Only in 43 out of 136 patients (31.6%) who were on tocolytic drugs, received AC. The schedule and doses were correct. According to the inquiry, 48/68 (0.71) of obstetricians considered AC as definitively useful, although only 22/68 (0.32) as very effective. The knowledge level was acceptable, except in signaling with precision the neonatal problems that could be prevented, as well as the lowest gestational age for use the AC. CONCLUSIONS: We considered the use of AC in our institution as suboptimal. We found that the reasons for not using them were not valid in most cases. Physician's attitudes to AC are positive but it is not reflected on their clinical practice. It is necessary a strong promotion of the usefulness of the AC for the preterm infant. PMID- 11006644 TI - [Impact of prenatal steroids on morbimortality of eutrophic premature newborns]. AB - Although there is evidence that the use of prebirth steroids promote maturity of premature newborn, in the United States of America it has been estimated that only 50% of the women who have had a premature delivery (with babies with weight between 501 and 1500 g) have been treated with steroids. We analyzed the morbimortality and the need of assisted ventilation in 91 premature newborns with gestational age of less than 34 weeks and weight of 1,500 g or less. They were divided in two groups: one group included 50 newborns whose mothers received at least one full course of steroids and the other group included 41 newborns whose mothers did not receive it. Newborn pathologies, the need of assisted ventilation and the mortality were higher in the group that did not receive prebirth steroids. We conclude that the use of prebirth steroids diminishes the morbimortality of the premature newborn. PMID- 11006645 TI - [Flexible vaginal cast covered with oxidated cellulose. Surgical option in neovagina for patients with Rokitansky-Kuster-Hauser syndrome]. AB - The alterations in differentiation of the mullerian derivatives and urogenital sinus are a frequent cause of demanding attention in the gynecologist office. Vaginal agenesis and hypoplasia are the representative events of alterations of human sexual differentiation. Handling such conditions is in direct relationship to the magnitude of the anatomical damage and the biopsychosocial consequences of such pathologies. From historical perspective, several surgical techniques of neovagina to correct such alterations have been described. We are presenting our experience in surgical treatment of vaginal agenesis, as well as the aesthetic functional outcomes obtained from a highly specialized hospital of the Health Sector in Mexico City. Our patients attended the gynecologist office mainly because of primary amenorrhea, anatomical alterations and sexual dysfunction. Tbe age range varied between 19 and 28 years. Basic laboratory tests were conducted. Urography, genetic studies, and a laparoscopic procedure were included. All the women received psychological support. The diagnosis in all the cases was Rokitansky Kuster-Hauser Syndrome with agenesis of the 2 proximal thirds of vagina. A neovagina assisted by laparoscopy using a technique previously modified was conducted. In such technique we use a flexible vaginal mould and oxide cellulose to prevent fibrosis and surgical retraction. Cosmetic outcome was satisfactory and dysfunction sexual was all improved. We conclude that our procedure that includes the neovagina assisted by laparoscopy and the use of a vaginal mould and oxide cellulose is a surgical option in those patients presenting with vaginal agenesis and hypoplasia. This procedure improves the aesthetic and sexual performances that are profoundly altered in such patients. PMID- 11006646 TI - [Risk factors for cesarean section: epidemiologic approach]. AB - The increase in frequency of cesareans that has been noted through 70's, not diminished--like it was expected--perinatal morbidity and mortality. The most important indications to cesarean are distocias, previous cesarean and fetal stress. In 1998 frequency of cesarean deliveries in our hospital was 35% of the pregnancy attended. The claim of this study was to determine risks factors to cesarean in our hospital. A case-control study was performed, selecting 165 cases (cesareans) and 328 controls (via vaginal). It was determined OR of the risks factors and atribuible fraction. Data were analyzed by X2. The most important indications to cesarean delivery were: distocias (39%, n = 64); previous cesarean (23%, n = 41) and fetal stress (11%, n = 21). There was not significative differences in age, height and rupture membrane time in both groups. History of cesarean delivery gave major risk to another surgical intervention (OR = 12.7, p = < 0.0001, atribuible fraction 92%). Nuliparous (OR = 6.6, p < 0.00000, atribuible fraction 85%), second gestation (OR = 1.8, p = 0.002) or history of abortion (OR = 1.8, p = 0.04) were factors mainly associated to cesarean delivery. We concluded that the precise 'medications of this surgical intervention specially in nuliparous or previous cesarean delivery cases must be replanteated to diminish its elevated frequency. PMID- 11006647 TI - [Prognosis factors associated with the progression of preeclampsia to eclampsia]. AB - This study was undertaken to determine the prognosis value of laboratory and clinical findings in the progression of preeclampsia to eclampsia. Nausea and vomiting and glucose level > 105 mg/dL, serum creatinine level > 1.0 mg/dL, aspartate aminotransferase level > 35 IU/L, alanine aminotransferase level > 40 IU/L and lactate deshiydrogenase level > 450 IU/L can be used to estimate the risk for the advancement to eclampsia. This information could be helpful to the clinician for management purposes. PMID- 11006648 TI - [Obesity, arterial hypertension, metabolic disorders, and polycystic ovary syndrome]. AB - OBJECTIVE: To determine the prevalence of central and general obesity, arterial hypertension, and alterations in cholesterol, triglycerids, glycaemia and uric acid levels in patients with PCOS and to compare them with women who do not present this syndrome. To establish the possible association of this with the PCOS. MATERIALS AND METHODS: This is a transversal analytic study. It includes fifty-five cases of PCOS and fifty-five healthy controls from gynecologic and familiar medicine consultation during the period from May 2nd to November 30th, 1999. It excludes patients with the Cushing disease, congenital suprarenal hyperplasia, pregnant women or women under hormonal treatments until three months prior the beginning of the study. The diagnostic of PCOS was established when the patient presents three of the following characteristics: menstrual disturbances, hirsutism, a LH/FSH relation > or = than 1.5 and ultrasound with polycystic ovaries. Their weight and height was taken in order to calculate the body mass index (BMI) and the waist/hip index (WHI) Blood samples in fasting where obtained to determine the FSH, LH, cholesterol, triglycerids, glycaemia and uric acid levels. Descriptive statistics where applied as well as the SPSS software along with the Saphiro-Francia test, the Mann Whitney test and logistic regression. RESULTS: The age of the patients with PCOS varied between 18 and 39 years old, average 26.6 years old and that of the controls from 19 to 39 years old, average 28.2 years old, (p = 0.165). The prevalence of general obesity (BMI > or = 30) was of 50.9% in the group of PCOS and of 18.2% in the group of controls. The systolic hypertension (BP > or = 140 mm Hg) was of 9% against 7.3%. A diastolic hypertension (BP > or = 90 mm Hg) of 25.5% against 7.3%. A hypercholesterolemia (> or = 220 mg/dL) of 30.9% against 10.9%. A hypertriglyceridemia (> or = 160 mg/dL) of 30.9% against 16.4%. A hyperglycaemia(> or = 115 mg/dL) of 5.5% against 3.6% and a hyperuricemia(> or = 6.5 mg/L) of 23.6% against 3.6%. The OR calculated through cross products showed that patients with PCOS have the possibility of presenting obesity, hypercholesterolemia, hypertriglyceridemia, hyperuricemia and diastolic hypertension, 4.6, 3.7, 2.3, 8.2 and 4.4 times more than the controls. The comparison of the average of the variables of both groups showed significant differences (p < 0.5) for the BMI, WHI, systolic BP, cholesterol and uric acid. The multiple logistic regression showed a consistent association between PCOS and hyperuricemia (MR = 5.23, = 1.4-26.26), obesity (MR = 3.26, CI = 1.33-8.01) and hypercholesterolemia (RM = 3.04, CI = 1.02-9.08). CONCLUSIONS: The study showed a higher prevalence of obesity, hypercholesterolemia, hypertriglyceridemia, hyperuricemia and diastolic hypertension in patients with PCOS than in the controls. No such thing for hyperglycaemia and systolic hypertension. Nevertheless, there are significant differences between both groups related to the BMI, WHI, systolic hypertension, hypercholesterolemia and hyperuricemia. A consistent association among the PCOS and hypercholesterolemia, obesity and hyperuricemia was found. PMID- 11006649 TI - Physicians' greatest enemy--complacency! PMID- 11006650 TI - Epidemiology of neural tube defects, Hawaii, 1986-1997. AB - Neural tube defects (NTDs) in Hawaii between 1986 and 1997 were examined using data from a statewide birth defects surveillance system. The prevalence increased significantly over the twelve-year period. NTD prevalence did not appear to vary by place of residence. The relationship of type of defect, maternal age, and infant/fetus sex was similar to that reported in the literature. PMID- 11006652 TI - Emphysematous pyelonephritis. AB - Emphysematous pyelonephritis is a rare, severe, necrotizing form of renal infection characterized by the presence of gas within the renal parenchyma or perinephric space. In patients suspected of emphysematous pyelonephritis, computed tomography scan should be done promptly. Based on the available data and this case, surgical intervention appears to be the preferred treatment. PMID- 11006651 TI - Alcohol use in Hawaii. AB - This article provides a review of the existing literature on alcohol use in Hawaii (i.e., epidemiology, reasons for use, associated problems, and intervention) and offers clinical implications of the findings and suggestions for further areas of research. In general, Caucasians, Hawaiians, younger Filipinos, males, adolescents, young adults, and those with lower educational attainment were found to be at higher risk. Overall, Hawaii's rates were either comparable or lower than those for the entire United States. Factors associated with different rates of alcohol use included accessibility, ability to resist offers, parent use and sanctions, peer influence and use, attitudes and beliefs (e.g., perceived normal drinking, dangerousness), religious affiliation, social occasions, and school intervention. Variable rates and trends in help-seeking behaviors, treatment admissions, and treatment utilization reflected the socio cultural diversity in Hawaii. Perceived effectiveness of different treatments were generally consistent across ethnic groups, but did not necessarily represent actual efficacy. There is a clear need for additional prevention, screening, and intervention programs in Hawaii, including socio-culturally appropriate ones, as well as a need for further research. PMID- 11006653 TI - Symptomatic human rotavirus subgroups, serotypes & electropherotypes in Hyderabad, India. AB - A total of 352 stool specimens obtained from children under 2 yr of age with acute diarrhoea, between January 1998 and March 1999, were screened for the presence of rotavirus by RNA-PAGE. Symptomatic human rotaviruses were detected in 57 of 352 (16.19%) specimens by RNA-PAGE. These 57 samples were tested for rotavirus double stranded RNA pattern and among these, 46 samples were tested for subgroup and serotype specificities. Among the 46 strains tested, 29 strains were found to be subgroup II and remaining 17 strains were subgroup I, indicating that subgroup II strains are more predominant than subgroup I strains. Subgroup I and II strains were circulating concurrently throughout the study period. Seventeen strains with 'short' RNA pattern and subgroup I specificity could not be assigned as serotype 2 strains as they exhibited cross-reactivity to MAbs specific for more than one serotype. Of the 29 subgroup II strains with 'long' RNA pattern, 16 (55.17%) were serotype 1, 8 (27.58%) were serotype 4. Five (17.24%) showed dual reactivity to serotypes 1 and 3. Our results indicated that serotype 1 and G2 like strains are predominant in Hyderabad. None of the virus strains showed an unusual RNA pattern. PMID- 11006654 TI - Detection of faecal leucocytes & erythrocytes from stools of cholera patients suggesting an evidence of an inflammatory response in cholera. AB - BACKGROUND & OBJECTIVES: Detection of faecal leucocytes and RBCs in stool samples of cholera patients has been reported in a small number of studies. This study extends these observations by examining stool samples of cholera patients in Calcutta. METHODS: Out of 1562 diarrhoeal stool samples, Vibrio cholerae was isolated in 266 cases. Stool samples obtained were examined microscopically within two hours of collection. RBCs and faecal leucocytes were examined by normal saline and methylene blue stain. Stool culture was performed using selective and differential media for isolation of V. cholerae. RESULTS: Among 266 cholera patients, RBCs was detected in 58 per cent and faecal leucocytes in 88 per cent respectively. The extent of the changes correlated with clinical severity. INTERPRETATION & CONCLUSION: This study showed the presence of RBCs and faecal leucocytes in stools of patients of cholera caused by V. cholerae 01 and 0139 which indicates some inflammatory changes in the gut mucosa. Further study is required to elucidate the inflammatory mechanism involved in the underlying process(es). PMID- 11006655 TI - Role of the male partner in the lower genitourinary tract infection of female. AB - We studied the relationships of selected microbes and the role of consorts in the causation of vaginal discharge which may be due to cervicitis or vaginitis. A total of 93 consecutive patients in the reproductive age group with symptoms of vaginal discharge along with their sexual partners were studied. Samples were collected from the cervix and posterior fornix of the female patients and from the urethra and sub-prepucial area of the male partner for culture of Neisseria gonorrhoeae, Gardnerella vaginalis, Mycoplasma hominis, ureaplasma, candida, aerobic and anaerobic organisms. Apart from cultures, KOH and Gram stain of the discharge were made. Predominant pathogen isolated was Ureaplasma urealyticum from 40 (43.01%) females and 23 (24.7%) males. The next common pathogenic organisms isolated were candida species from 11 (11.8%) females and 5 (5.4%) males and Chlamydia trachomatis in 3 (3.2%) females and 1 (1.1%) male. Various organisms were more frequently isolated from the 29 of 43 (67.4%) couples who had had sexual intercourse 7 days prior to the recruitment as compared to 14 of 43 (32.6%) who had had coitus more than 7 days prior to their recruitment. This may be due to the spontaneous disappearance or decrease in the number of organisms to the level that they could be detected by culture. In our study, 6 (6.5%) of male partners carrying pathogenic organisms were asymptomatic, indicating that their screening and treatment is vital. PMID- 11006656 TI - The effects of clofazimine, niclosamide & amphotericin B, on electron transport of Leishmania donovani promastigotes. AB - BACKGROUND & OBJECTIVES: The study was undertaken to explore the locus of interaction of clofazimine and niclosamide which showed substantial growth inhibition property in Leishmania donovani promastigotes. METHODS: The uptake of final electron acceptor oxygen and 2,6-dichlorophenolindophenol (DCPIP) reduction in the electron transport chain were measured by constant volume Warburg respirometer and monitoring absorbance at 600 nm, respectively. Irreversibility of O2 uptake inhibition by clofazimine and niclosamide was determined by dilution of cell suspension followed by centrifugation. RESULTS: Clofazimine and niclosamide showed their minimum inhibitory concentration (MIC) at 33 and 150 micrograms/ml, respectively. Oxygen uptake inhibition by clofazimine and niclosamide was not reversed by removal of the drug by centrifugation. Rotenone, a potent inhibitor of mammalian electron transport chain showed no inhibition on the electron transport chain of L. donovani promastigotes. Cyanide at 1 mM concentration showed partial inhibition in L. donovani promastigotes. Oxygen uptake and DCPIP reduction by L. donovani promastigotes were highly sensitive to sulphhydryl group inhibitors. Strong inhibition of oxygen uptake (80-100%) by L. donovani promastigotes was achieved by clofazimine, niclosamide and amphotericin B. Amphotericin B failed to inhibit DCPIP reduction by L. donovani promastigotes, whereas DCPIP reduction was inhibited by clofazimine and niclosamide, respectively. INTERPRETATION & CONCLUSION: DCPIP reduction was mediated by transplasma membrane electron transport as evidenced by its inhibition with membrane impermeable quinone 1,2-naphthoquinone-4-sulphonic acid (NQSA). Transplasma membrane electron transport requires b-cytochromes and sulphhydryl groups for its function and was inhibited by clofazimine and niclosamide. PMID- 11006657 TI - Air pollution in Calcutta elicits adverse pulmonary reaction in children. AB - BACKGROUND & OBJECTIVE: Pulmonary responses of children chronically exposed to ambient air pollution in Calcutta have been investigated. METHODS: A total number of 153 children from Calcutta and 116 from rural West Bengal in the age group of 6-17 yr were included in this study. Respiratory symptom complex, sputum cytology and micronucleus (MN) count of buccal epithelial cells were evaluated. Blood smears were examined for WBC differential count and RBC morphology. RESULTS: Marked rise in respiratory symptoms (43% in urban vs 14% in rural) and sputum alveolar macrophage (AM) number was observed in urban children compared to their rural counterparts (14.2 +/- 1.4 AM/hpf vs 6.7 +/- 1.4 AM/hpf, mean +/- SE, P < 0.001). The urban group also demonstrated increased numbers of neutrophils, eosinophils and iron-laden AM in their sputum. Besides, buccal epithelial cells of urban children exhibited higher MN frequency than their rural counterparts (0.22 vs 0.17%, P < 0.05). While sputum neutrophilia and eosinophilia suggest inflammatory and allergic lung reactions, elevated MN count is indicative of greater genotoxic effect on the exposed tissues of urban children. Hypochromic red cells in peripheral blood smear was a common finding in both urban and rural groups, but eosinophils and monocytes were present in elevated frequencies in the rural children. INTERPRETATION & CONCLUSION: The study demonstrated that children inhaling grossly polluted air of Calcutta suffer from adverse lung reactions and genetic abnormality in the exposed tissues. PMID- 11006658 TI - Dimensions & shape of the trachea in the neonates, children & adults in northwest India. AB - Measurements of the trachea and the two main bronchi including the subcarinal angle and the angles of the bronchi with the vertical were taken in specimens obtained from 370 adults, 60 children and adolescents up to 17 yr and 27 dead newborns. These measurements showed a gradual increase with age up to a certain point (often 31-35 yr in males and 26-30 yr in females) and stabilised thereafter. There was no sexual dimorphism in measurements till the age of 17 yr and thereafter, the measurements were greater in males than in females (P < 0.001). In adults the body weight and supine body length had a significant correlation with certain measurements of the trachea. The subcarinal angle and the angles of the two main bronchi with the vertical were variable but the angle of the right bronchus with the vertical was always found to be smaller than that of the left. The incidence of different shapes of the trachea at transversely cut upper end was also studied. A U-shaped trachea was present in 33.2 per cent male adults but was not seen in females. This may have some medico-legal importance. PMID- 11006659 TI - Should infantile hypertrophic pyloric stenosis be managed in specialist centres? PMID- 11006660 TI - Preoperative anaesthesia information--what do patients need to know? PMID- 11006661 TI - Thrombolytic therapy in acute myocardial infarction: Third Irish Working Party Consensus. Irish Thrombolysis Concenus Group. PMID- 11006662 TI - Infantile hypertrophic pyloric stenosis in a regional centre. AB - BACKGROUND: It has been suggested that only specialist paediatric surgeons should manage infantile hypertrophic pyloric stenosis (IHPS). AIM: The aim of this retrospective study was to ascertain whether the majority of these infants might be managed in a well-equipped regional centre. METHODS: Using the Hospital Inpatient Enquiry database, all cases of IHPS within a single administrative health region were identified over a six-year period. A chart review was performed to obtain demographic and clinical information for each patient. Reports from the Central Statistics Office were used to obtain data on population and live births for the region. RESULTS: Seventy patients with IHPS were identified from this region from 1991 to 1996, 63 (90%) of which were treated in the region. Of the remaining seven, four were referred directly to specialised paediatric surgical hospitals because of prematurity (n = 1), low birth weight (n = 1), capillary haemangioma (n = 1) and severe metabolic derangement (n = 1) while three were assessed and treated in general hospitals outside the region. Of the 63 infants undergoing pyloromyotomy in this region, the duodenal mucosa was breached in four (6%); there were wound complications in three (5%); and one required a re-pyloromyotomy. The mean postoperative stay was eight days (range 2 42 days). CONCLUSION: The majority of infants with IHPS may be safely managed in regional centres with transfer to specialist paediatric centres reserved for 'high risk' cases. The management of IHPS at a regional level has important implications and presents opportunities for surgical training. PMID- 11006663 TI - Management of head injury in a regional hospital. AB - BACKGROUND: Concern about delay in the treatment of serious head injury may result in unnecessary transfer to neurosurgical units for scanning and neurosurgical assessment. AIMS: This study assessed the management of head injuries in a regional hospital with computed tomographic (CT) and 'image-link' facilities allowing instantaneous transmission of CT scans to the regional neurosurgical unit for specialist assessment. METHODS: A retrospective study was carried out of all head injuries presenting to Limerick Regional General Hospital in a single year. Data gathered included mechanism of injury, mode of transfer, requirement for admission and length of stay. Neurological status was assessed using the Glasgow Coma Scale (GCS) and functional status using the Glasgow Outcome Scale. RESULTS: Between January and December 1995, 1,564 patients presented with head injuries to the accident and emergency (A/E) department of Limerick Regional General Hospital. Twenty (1%) were dead on arrival, 12% required hospital admission and the remainder were discharged after assessment. Seventy-six per cent were males and 74% under 40 years of age. Among the 194 patients requiring hospital admission, 14% had a skull fracture and 22 had a severe head injury. CT brain scans were performed in 43 patients and were abnormal in 42%. On the basis of CT and clinical findings, six patients were transferred immediately for a neurosurgical procedure and one was transferred later following clinical deterioration. CONCLUSIONS: A CT scan and image-link facility permitting remote neurosurgical advice allows the majority of patients with head injury to be safely managed in well-equipped regional units without onsite neurosurgical expertise. PMID- 11006664 TI - Regional vascular surgical units: a practical means of providing specialist services. AB - BACKGROUND: A comprehensive regional emergency and elective vascular surgical service was established in Limerick Regional General Hospital between 1994 and 1996 following the appointment of three surgeons with vascular training. AIM: To compare vascular surgical activity before and after the establishment of a regional vascular service. METHODS: This study used local and national computerised databases, clinic letters, theatre registers and radiology records to compare vascular surgical activity before (1992) and after (1997) this service was established. RESULTS: Total outpatient activity increased almost six-fold and total number of inpatient vascular procedures (including radiological) increased from 146 to 432, but the venous proportion declined from 70% to 36%. The number of major arterial procedures increased from 37 to 165 in 1997 including 10 carotid endarterectomies, 24 aortic reconstructions and 54 lower limb reconstructions representing rates of 3/100,000, 8/100,000 and 17/100,000 population respectively. CONCLUSION: As substantial changes in disease patterns are unlikely, these data indicate that patients previously diverted elsewhere for therapy are now cared for within the health board region and that further increases in workload may be expected. We suggest that these data mandate the reallocation of resources to fund appropriately staffed and audited regional vascular units. PMID- 11006665 TI - Prevalence and aetiology of leg ulcers in Ireland. AB - BACKGROUND: The prevalence of leg ulcer disease in Ireland has been poorly documented. AIMS: This study aimed to investigate the aetiology and prevalence of leg ulcers in one health district. METHODS: All patients receiving healthcare for an active leg ulcer in the Mid-Western Health Board (MWHB) region of Ireland (population: 317,069) were identified in a defined two-month period. A cross sectional survey of all healthcare workers providing care to patients with leg ulceration was carried out. Patients with leg ulcers of uncertain cause were invited for follow-up assessment to establish the underlying cause. RESULTS: There were 389 patients with leg ulcers with a mean (standard deviation [SD]) age of 72.3 (11.1) years. The prevalence was 0.12% but it was 1.03% in patients aged 70 years and over. Women were twice as likely to be affected. Venous disease accounted for 81% of ulcers, and arterial disease for 16.3%, while ulceration due to diabetic neuropathy and rheumatoid vasculitis was unusual. CONCLUSION: Leg ulcers are an important source of morbidity in our ageing population. Effective treatment programmes could diminish the impact of this debilitating disease on the health service. PMID- 11006666 TI - Patients' knowledge of perioperative care. AB - BACKGROUND: Patient knowledge of perioperative care, particularly the role of the anaesthetist, appears limited. AIM: This study investigated patients' knowledge of common medical terms, of their surgical care, of the role of the anaesthetist, and their preoperative fears and concerns about postoperative pain. We examined the changes in their knowledge base as a result of the pre-anaesthetic visit and their inpatient stay. METHODS: Three hundred patients were surveyed on three separate occasions, before a routine pre-anaesthetic visit, two to three hours after this visit and on the day of discharge from hospital. RESULTS: Patients' knowledge of medical terms, their surgical procedure, and the role of the anaesthetist in their perioperative care were limited. There was little change following either the pre-anaesthetic visit or postoperative convalescence. Misconceptions, about such issues as postoperative pain, appeared to increase by the end of their hospital stay. Male patients, older patients and patients in lower socio-economic groups had poorer knowledge. A significant proportion of patients remained unaware that the anaesthetist was medically qualified. CONCLUSIONS: Patients' knowledge of perioperative care is limited, with very little change during hospital stay. Novel educational approaches may be required to increase basic medical knowledge. PMID- 11006667 TI - The management of ureteric calculi without extracorporeal shock-wave lithotripsy. AB - BACKGROUND: Extracorporeal shock-wave lithotripsy (ESWL) is a relatively new technological adjunct in the treatment of renal calculi, but availability is limited. AIM: The aim of this study was to assess the outcome of ureteroscopic procedures in a unit without on-site ESWL facility. METHODS: A retrospective study of all cases of ureteric calculi presenting to this hospital during the period 1991 to 1997 was performed. RESULTS: One hundred and thirty five patients with a mean age of 46.8 years were evaluated. Ninety-four (69.5%) had their stone successfully manipulated and/or extracted at ureteroscopy. Using strict criteria, 41 patients (30.5%) had failed ureteroscopies. Fourteen (10.4%) suffered complications related to their ureteroscopy, thirteen had their complications treated conservatively and three (2.3%) needed open procedures. CONCLUSION: Ureteroscopy has a high success rate and low morbidity. We continue to advocate ureteroscopy in the initial management of ureteric calculi. PMID- 11006668 TI - Profile of attendance at a maternity hospital emergency room. AB - BACKGROUND: Direct presentations to secondary care that could have been managed in primary care have been the subject of much study. Social factors may influence the decision to present directly to secondary care services. AIM: To determine the profile and primary care service use of patients attending a maternity hospital emergency room (ER). METHODS: A self-administered, questionnaire based, cross-sectional study was performed over a period of one month. RESULTS: Of 350 patients attending the ER, 343 (98%) completed a questionnaire. Mean age of patients attending was 28 years (range 16-65). Thirty seven per cent were eligible for free medical care and 39% belonged to socio-economic group six. Fifty nine per cent attended outside normal hours, 31% had a female general practitioner (GP) and 38% were referred by a GP to the ER. Twenty eight per cent had seen their GP in the previous three days, 31% had consulted a GP with their current problem and 58% felt "comfortable" or "very comfortable" attending their GP with a gynaecological complaint. CONCLUSIONS: This study indicates that the service is responsible for a considerable workload, with a low level of use of primary care services by those attending. PMID- 11006669 TI - Need-based waiting lists for hip and knee arthroplasty. AB - BACKGROUND: Need-based waiting lists have been shown to offer advantages over traditional time-based waiting lists. This study investigated the effect of their introduction in our orthopaedic practice. METHODS: Each patient on the waiting list for total hip replacement (n = 240) and total knee replacement (n = 98) was invited to attend a dedicated assessment clinic. At the clinic each patient had a joint score (Harris Hip Score or American Knee Society Score) calculated. Patients with the lowest joint scores were moved to the top of the waiting lists. Scores were repeated postoperatively. RESULTS: Validation of the hip replacement and knee replacement waiting lists reduced them by 20% and 11% respectively. The efficiency and transparency of the service were increased. CONCLUSION: Need-based waiting lists are easily introduced and offer significant advantages over time based waiting lists. We advocate their introduction in other centres. PMID- 11006670 TI - A comparison of outcome scores in os calcis fractures. AB - BACKGROUND: Of over 20 different scoring systems to evaluate outcome following calcaneal fracture, the Maryland Foot Score has broad current acceptance. A general health survey, the Short Form 36 (SF 36) has also been used. AIMS: The study compared two scoring systems for assessing the outcome of open reduction and internal fixation of displaced intra-articular calcaneal fractures. METHODS: Over a four-year period, 31 displaced intra-articular calcaneal fractures were categorised by the Sanders classification and treated by open reduction and internal fixation. Outcome was assessed by the Maryland Foot Score and by the Short Form 36 (SF 36) general health questionnaire. RESULTS: Thirty-five per cent of fractures were Sanders class 2, 57% were class 3 and 8% were class 4. The average follow-up was 2.5 years (range 1-4 years). Using the Maryland Foot Score, seven fractures had an excellent result, 13 had a good result, six had a fair result and one was a failure. There was a correlation between pain (coefficient = 0.780, p < 0.001) and physical function (coefficient = 0.638, p < 0.001) with the appropriate sections of the SF 36. CONCLUSION: The Maryland Foot Score measures what it attempts to measure and therefore it has content validity for pain and physical function. PMID- 11006671 TI - Assessment of hepatitis C infection in injecting drug users attending an addiction treatment clinic. AB - BACKGROUND: Injecting drug users represent a high risk group for hepatitis C (HCV) infection. Currently, screening of this group for HCV is inconsistently implemented. AIM: We designed a HCV assessment algorithm and sought to determine the frequency with which injecting drug users completed the assessment process. METHODS: Prospective study of a HCV assessment algorithm in the setting of a specialist outpatient addiction treatment clinic. Participants consisted of consecutive new attenders over a six-month period with a history of injecting. RESULTS: Only 21 (18%) of 119 patients reached a satisfactory endpoint of assessment. Forty-eight injectors were tested for antibody to HCV, of whom 26 (54%) tested positive. Thirteen of those with positive test results were no longer attending when the test result became available. Only four of the 19 patients who were referred, attended the on-site hepatology clinic. CONCLUSIONS: Thorough screening of injecting drug users for HCV within one treatment service is difficult. There is a need for explicit policies on this issue involving co operation between primary care providers and addiction services and hepatology services. PMID- 11006672 TI - Methadone maintenance in general practice: impact on staff attitudes. AB - BACKGROUND: The evaluation of a structured protocol for the discharge of stabilised patients on methadone maintenance to general practice provided an opportunity to evaluate the impact on the attitudes of general practitioners (GPs) and practice staff. AIM: To assess attitudes, expectations and experience among GPs and practice staff before the introduction of structured methadone maintenance and six months after its introduction. METHODS: A postal questionnaire was sent to 31 GPs and 23 receptionists in 23 Dublin general practices before the patient's first visit and six months later at the end of the study period. Outcome measures were staff attitudes, incidence of disruption, perceived difficulties in providing care and in prescribing methadone, and stress levels. RESULTS: There was a generally positive attitude to provision of methadone in general practice for stabilised patients, although it was not anticipated to be problem free. Following six months involvement attitudes were similar; stress levels were unchanged, but fewer GPs anticipated problems in delivering the service. All continued to participate in the scheme. CONCLUSION: GPs and receptionists in this sample had mixed views about methadone maintenance which were unchanged by six months experience of the service. The study illustrates important issues in the recruitment and support of general practice in meeting this need. PMID- 11006673 TI - Population characteristics of Irish Helicobacter pylori isolates: a tRNA associated locus. AB - BACKGROUND: A strain-variable transfer RNA-associated-locus (trl) was present in 50% of Irish Helicobacter pylori (H. pylori), isolates and did not correlate with the origin of the isolates. AIM: To associate a particular genotype or phenotype to trl status in H. pylori by further screening the isolates from the original study for the presence of known genotypic and phenotypic characteristics. METHODS: Forty two clinical isolates were screened for the presence of the cagA, vacA, iceA1 and vapD genes by Southern or DNA dot blot analysis. Western blot analysis was performed using antibodies to CagA, VacA, Lewis X (Le(x)) and Lewis Y (Le(y)). Plasmids were identified by the alkaline lysis method. RESULTS: The cagA gene was present in 29 (69%) of isolates screened and 21 (50%) produced the CagA protein. The vacA gene was detected in all of the isolates while VacA was expressed in 71.4%. The iceA1 and vapD loci were detected in 73.8% and 71.4% respectively. Le(x) was expressed in 42.9% and Le(y) in 38.1% of the isolates. Expression of both Lewis antigens was detected in 7.1% while in 30.9% neither antigen was detected. Plasmids were present in 47.6%. There was no association between the trl status of isolates and any of the above. There were no significant associations between the phenotypic and genotypic characteristics studied and peptic ulcer disease or non-ulcer dyspepsia. CONCLUSION: The strain variable tRNA-associated locus is independent of the vacA/VacA, cagA/CagA, Lewis X, Lewis Y, iceA1, vapD and plasmid status in the population of Irish H. pylori isolates studied. PMID- 11006674 TI - Acute forearm compartment syndrome: report of three cases and a review of the literature. PMID- 11006675 TI - The Irish Journal of Medical Science--a historical outline. PMID- 11006676 TI - Idiopathic mesenteric thrombosis following caesarean section. PMID- 11006678 TI - Intussusception in adults: a rare entity. PMID- 11006679 TI - The roots of youth violence. PMID- 11006677 TI - Unusual case of pilomatricoma. PMID- 11006680 TI - Reaching communities of children. PMID- 11006681 TI - Troubled kids, troubled times. PMID- 11006682 TI - Boys and their toys. PMID- 11006683 TI - Babes in arms. Juvenile access to guns in Minnesota. PMID- 11006684 TI - Striking back. PMID- 11006685 TI - Media violence. The harsh reality. PMID- 11006686 TI - Interactive media violence and children. PMID- 11006687 TI - Protecting youth from harm. PMID- 11006688 TI - Promising approaches to youth violence prevention. PMID- 11006689 TI - Beyond treating the wounds. The physician's role in preventing gun violence. PMID- 11006690 TI - Listening to rural Minnesota. PMID- 11006691 TI - Pediatric firearm injury in Minnesota, 1998. Fatal and nonfatal firearm injuries among Minnesota youth. AB - OBJECTIVE: Minnesota hospitals began to assign E-codes (external cause of injury) to hospital discharge data in 1998. It is now possible to describe the epidemiology of medically treated firearm injury (FI) cases in Minnesota by combining hospital discharge data with injury data from other sources. This population-based investigation provides a preliminary epidemiologic description of fatal and nonfatal firearm injuries in children and adolescents in Minnesota during 1998. METHODOLOGY: Pediatric firearm injury cases were identified from the Minnesota Department of Health's Minnesota Trauma Data Bank, a population-based data system for injury surveillance in Minnesota. To qualify for the study, the patients had to be Minnesota residents, younger than 20 years old at the time of injury, and injured by a firearm in 1998. RESULTS: The 175 cases identified yielded on overall FI rate of 12.5/100,000, a mortality rate of 2.4/100,000, and a nonfatal to fatal ratio of 4.1:1. Eighty-five percent of patients were male; 15% female. The largest proportion of firearm injuries were assault-related (45%), followed by unintentional (34%), and self-inflicted (15%). Adolescents aged 15-19 years accounted for 79% of the injuries, and children 14 or younger, 21%, yielding rates of 37.6/100,00 and 3.5/100,000, respectively. Twenty-two percent of 15-19-year-olds and 11% of children 0 to 14 died, yielding mortality rates of 8.1/100,000 and 0.4/100,000, respectively. Where race was documented, whites represented 53% of the cases; African Americans, 32%; Native Americans, 8%; and Asian/Pacific Islanders, 7%; yielding race-specific rates of 4.1/100,000, 54.3/100,000, 30.5/100,000, and 12.1/100,000, respectively. Sixty-six percent of the patients were residents of the Minneapolis/St. Paul metro area and 33% were residents of Greater Minnesota, yielding rates of 16.0/100,000 and 8.6/100,000, respectively. In the Twin Cities metro area 24% of injury cases were unintentional, 5% were self-inflicted, 65% were assaultive, and 6% were undetermined. In Greater Minnesota 53% were unintentional, 33% were self inflicted, 9% were assaultive, and 5% were undetermined. CONCLUSIONS: Minnesota's 1998 rates for firearm injury were less than half the 1997 national rate. The state's FI rates show significant disparities by race and region of residence. More FI data will have to be collected over more years to describe and identify trends and multifactor relationships. Policymakers need to ensure that firearm injury continues to be monitored and assessed in Minnesota. PMID- 11006692 TI - Firearm-related suicide in Minnesota. A picture of morbidity and mortality. PMID- 11006693 TI - Calculating the costs of gun injuries. A new methodology. AB - A new method for assessing the costs of gun injuries to a health system examines data on paid amounts, comprehensive medical expenses, and expenses over time. The authors extracted claims using injury diagnosis codes from billing forms and medical charts. The study demonstrates that a claims database can be used to accurately measure health care costs associated with gun injuries. The study is the first to include gun-related injuries treated in ambulatory care settings and to track actual payments over time. PMID- 11006694 TI - [Recurrent thoracic pain without findings. Let the endoscope evaluate the situation!]. PMID- 11006695 TI - [Space-occupying lesion in the liver? Don't immediately refer for expensive CT examination!]. PMID- 11006696 TI - [More rapid recovery after fracture. When can ultrasound accelerate healing?. Interview by Petra Eiden]. PMID- 11006697 TI - [Sports or hormones for aging men? When Adam suffers from PADAM (partial androgen deficit of the aging man)]. PMID- 11006698 TI - [After-care in breast carcinoma. Medical examination and mammography compared]. PMID- 11006699 TI - [New therapeutic approaches in advanced breast carcinoma. Palliative measures are increasingly more tolerable]. AB - Advances in palliative chemotherapy are based on the development of new substances or new treatment strategies, such as, for example, high-dose chemotherapy. New cytostatic agents are developed with the aim of improving therapeutic efficacy and tolerability. Taxanes rapidly became part of standard treatment, and their combination with the anthracyclines represents the most effective treatment currently available. Liposomal encapsulation of anthracyclines can reduce their cardiotoxicity without loss of efficacy. Recent antimetabolites and the vinca alkaloid, vinorelbine, combine a good antitumoral effect with a favorable toxicity profile. The use of the monoclonal antibody, trastuzumab, has made specific immunotherapy possible for the first time. The role of high-dose chemotherapy remains uncertain, and further randomized studies are urgently needed. PMID- 11006700 TI - [Resection of the sentinel lymph node in breast carcinoma. Substitution for axillary lymph node excision?]. PMID- 11006701 TI - [With hypobaric pressure to diagnosis. Vacuum punch biopsy allows for minimally invasive sample study]. PMID- 11006702 TI - [Internet addresses on the main topic. Breast carcinoma]. PMID- 11006703 TI - [New guidelines for resuscitation. Mouth-to-mouth resuscitation and 100 compressions per minute]. PMID- 11006704 TI - [Human granulocytic ehrlichiosis. New tick bite disease lies in wait also in German forests]. AB - HGE is an uncommon zoonosis, transmitted to humans by ticks (Ixodes ricinus). Heavily exposed persons, such as foresters and forestry workers, are most at risk of infection. HGE usually runs an asymptomatic and self-limiting course. Symptoms include acute pyrexial illness, frequently accompanied by muscle pain, headache, nausea and vomiting. Treatment is with doxycycline/tetracycline or rifampicin. In a study in Hesse of 128 heavily exposed subjects, 19.5% had serum antibodies (IFA test) against HGE versus 4.4% in a control group. This difference is statistically significant. The prevalence of both symptoms and the seropositivity differ significantly between locations. HGE should be considered in the differential diagnosis of fever of unknown origin following tick bite exposure, at least in the North Hesse region. PMID- 11006705 TI - [Guidelines for lowering lipids are too infrequently observed. Results of a retrospective study of coronary heart disease patients]. AB - The aim of this retrospective analysis was to investigate adherence to treatment guidelines in the secondary prevention of coronary heart disease. 3720 CHD patients treated in 591 doctor's offices throughout Germany were investigated. End points were serum lipid levels at week-6, i.e. screening investigation, and 5 weeks after discontinuation of the lipid-lowering medication (week-1). 3383 of the 3720 (90.9%) patients had LDL-C levels > = 115 mg/dl, and 3563 (95.8%) > = 100 mg/dl. At week-6 mean LDL-C was 167.7 +/- 43.5 and mean total cholesterol was 258.8 +/- 47.8 mg/dl. 5 weeks after discontinuation of lipid-lowering treatment, mean LDL-C increased by 5.6%, and mean total cholesterol by 3.7% in comparison with baseline at week-6. 2346 (69.3%) of the patients with LDL-C > = 115 at week 6 did not receive any prior lipid-lowering medication. Also, patients receiving lipid-lowering medication demonstrated an insufficient lipid decrease (only 14.4% of all treated patients had LDL-C levels < 115 mg/dl, and only 6.8% had levels < 100 mg/dl. CONCLUSION: So far, lipid-lowering guidelines for the secondary prevention of CHD are not being adequately implemented. Appropriate action to remedy this situation (e.g. establishment of Disease Management Programs) is needed. The aggressive use of lipid-lowering drugs is a must if the goals of the treatment guidelines are to be met, and morbidity and mortality of CHD lowered. PMID- 11006706 TI - [Thrombosis ABC, 19: Lung embolism. When thrombosis becomes an emergency]. PMID- 11006707 TI - [Diagnostic quiz. Thoracic pain in a young woman. Acute posterior wall infarct]. PMID- 11006708 TI - [Senile macular degeneration. Photodynamic therapy stabilizes vision]. PMID- 11006710 TI - [Therapy of allergic conjunctivitis]. AB - Among the several eye diseases (or diseases involving the eye) based on hypersensitivity reactions, the most frequent is allergic conjunctivitis. Recently six types of allergic conjunctivitis/keratoconjunctivitis are distinguished: 1. seasonal, 2. perennial, 3. vernal, 4. giant papillary, 5. atopic and 6. of contact origin. Their treatment is generally local. In the most frequent seasonal ("hay fever") and perennial forms the elimination of the allergen or when it is impossible antihistamines (with or without vasoconstrictors), "weak" steroids or hyposensitisation are offered. In vernal and atopic keratoconjunctivitis mast cell stabilizers are the most effective, with special effect of lodoxamide in the vernal type. In giant papillary and contact allergic inflammations the elimination of the causative agent is the first method of choice. In resistant cases "strong" steroids, in extreme forms immunosuppressive, cytostatic and systemic treatment may become necessary. The paper gives a review of currently applied medicines (mainly eyedrops) and other methods of treatment, and includes therapeutic principles applying to various forms of allergic conjunctivitis. PMID- 11006709 TI - [Pathogenesis and types of neonatal diabetes]. AB - Studies on pathogenesis of neonatal diabetes may lead to better understanding of pancreatic beta-cell maturation and regulation of insulin secretion. The purpose of this study was to examine the clinical course, immunologic and genetic background of the transient and permanent form of neonatal diabetes. Clinical data (onset and duration of diabetes, C peptide levels, duration of insulin treatment, associated disorders) were collected, islet cell specific antibodies (islet cell antibody and glutamate decarboxylase antibody) were determined and genetic analyses (HLA DQA1-DQB1 typing and microsatellite mapping on chromosome 6) were performed in three patients with permanent and three patients with transient neonatal diabetes. None of the six patients had HLA DQ diabetes susceptibility alleles and most infants were negative for islet cell autoantibodies indicating that no pancreatic islet-cell specific autoimmunity exists in the two forms of neonatal diabetes mellitus. Complete paternal uniparental isodisomy of chromosome 6 has been identified in a transient neonatal diabetes case with macroglossia. The permanent neonatal diabetes cases and the other two cases with transient form did not possess this chromosome anomaly. It is concluded, that none of the two forms neonatal diabetes is of autoimmune origin. They have different genetic background and represent different disease entities. Transient neonatal diabetes is probably caused by alteration of expression of an imprinted gene on chromosome 6 which might play role in fetal growth and pancreatic beta-cell maturation and insulin secretion. Permanent neonatal diabetes is probably a more heterogenous phenotype, the cause of which remains to be clarified in the future. PMID- 11006711 TI - [Effectiveness of Lisinopril in the treatment of heart failure]. AB - A prospective study was performed in patients (30 M, 16 F, mean age of 56.0 +/- 9.2 [42-73] years) with congestive heart failure to assess the efficacy of lisinopril during a 16 weeks treatment period. Changes in clinical signs, functional capacity, blood pressure, heart rate, echocardiographic parameters, exercise duration, laboratory data and quality of life were measured. After a 2 week run-in period starting daily dose of study drug was 5 mg, and an increase of medication was considered at 4 weeks. At the end of the study mean daily dose of lisinopril was 15.1 +/- 6.2 mg. Improvement of NYHA status by 2 grades was observed in 4 cases (9%), by 1 grade in 24 cases (51%), there was no change in 17 cases (38%), and worsening was observed in 1 case (2%). During the study both systolic (p = 0.001) and diastolic blood pressure (p = 0.0006) decreased significantly, the changes in pulse rate were not significant. Left ventricular end systolic (p = 0.001) and end diastolic (p = 0.003) dimensions decreased, ejection fraction rose by 4.4% (p = 0.0002). One patient was removed from the study because of drug-induced cough. Comparison of all the laboratory data for pre and post-study periods did not reveal any significant difference. Patients treated with lisinopril improved significantly for clinical, haemodynamic, echocardiographic and quality of life parameters, with few adverse experiences, good tolerability and once-daily dose. PMID- 11006712 TI - [22q11.2 chromosome deletion and velo-cardio-facial syndrome in a patient with tetralogy of Fallot]. AB - Velo-cardio-facial syndrome includes the following clinical features: congenital heart anomaly, velo-pharyngeal malformations and dysmorphic facial features. In 80% of the patients microdeletion of the long arm of chromosome 22 can be detected. Many patients with DiGeorge syndrome have the same underlying chromosomal etiology, however the same deletion results in severe dysmorphic features, immune defect, hypocalcaemia and in severe mental retardation. In isolated heart defects deletion can be detected in about 15% of the patients. We report the detection of the microdeletion in a patient with tetralogy of Fallot, minimal dysmorphic facial features and normal psychomotor development. Early diagnosis of 22q11.2 chromosomal deletion is important for genetic counseling and further rehabilitation. We emphasize the importance of genetic screening in patients with isolated congenital heart defects. PMID- 11006713 TI - [Anniversary of the first brain tumor removal in Hungary -- remembering Mor Lipscher]. PMID- 11006714 TI - [Remembering a medical historian, Ferenc Kolos Mayer, on his birth centennial]. PMID- 11006716 TI - [Ethical problems of continuing medical education and self-education]. PMID- 11006717 TI - [Hungarian medical writings -- Castleman disease]. PMID- 11006715 TI - [Treatment of laryngeal constriction. By Professor Mihaly Groszmann of Vienna, 1875]. PMID- 11006718 TI - Does the type of judgement required modulate cue competition? AB - According to the comparator process hypothesis (Matute, Arcediano, & Miller, 1996), cue competition in the learning of between-events relationships arises if the judgement required involves a comparison between the probability of the outcome given the target cue and the probability of the outcome given the competing cue. Alternatively, other associative accounts (the Rescorla-Wagner model: Rescorla & Wagner, 1972) conceive cue competition as a learning deficit affecting the target cue-outcome association. Consequently, the comparator process hypothesis predicts that cue competition occurs in inference judgements but not in contiguity ones, for only the first type of judgement implicitly involves such a comparison. On the other hand, the Rescorla-Wagner model predicts cue competition in both inference and contiguity judgements, because it establishes no relevant role for the type of judgement in producing cue competition. In Experiments 1 and 2 we manipulated the relative validity of cues and the type of question (inference vs. contiguity) in a predictive learning task. In both experiments we found a cue competition effect, but no interaction between the relative validity of cues and the type of question, suggesting that the Rescorla-Wagner theory suffices to explain cue competition. PMID- 11006719 TI - Stimulus representations in human Pavlovian conditioning: implications of missing negative transfer across response systems. AB - Three Pavlovian conditioning experiments with human participants are reported, which investigated whether common or separate stimulus representations are involved in solving nonlinear discrimination tasks in different response systems. In our experiments we made use of a negative transfer effect between positive and negative patterning. Experiment 1 specified the conditions under which such a negative transfer effect occurs in human eyelid conditioning. Experiments 2 and 3 investigated whether a similar effect also occurs if two response systems--the eyelid and the skin conductance response system--are trained with trials of both types being randomly interleaved. The presence or absence of a negative transfer effect indicates whether or not the stimulus representations involved in the two conditioning processes overlap. The findings are discussed within the framework of a neuropsychological model of hippocampal function. The results suggest that the representations are distinct and thus support the idea of acquired equivalence and distinctiveness of stimulus representations. PMID- 11006720 TI - Rapid visual learning in the rat: effects at the 5-HT1a receptor subtype. AB - The 5-hydroxytryptamine1a (5-HT1a) receptor agonist 8-hydroxy-2-(di-n propylamino) tetralin (8-OH-DPAT; 0.15 mg/kg) impaired rats' rapid visual learning on a computerized maze. This treatment also increased decision time (DT) but the learning impairment was not necessarily a side-effect of slower responding because, in this task, responses made at long DT are more accurate than those at short DT. The selective 5-HT1a receptor antagonist WAY-100635 (0.3 mg/kg) was itself without effect on accuracy, but was effective in reversing effects of 8-OH-DPAT (on both accuracy and DT). Within problems (i.e., over the 40-60 trials of a single discrimination), performance was reduced by treatment with 8-OH-DPAT at all stages of learning. We conclude that this effect is mediated through the 5-HT1a receptor site (rather than through some other serotonergic receptor site or non-specific mechanism) as it was reversible by treatment with WAY-100635. Although it could still arise from behaviourally non specific effects, the performance deficit finds its best account in terms of the psychological processes necessary to visual learning. Its reversal with WAY 100635 offers support to the hypothesis that 5-HT1a receptor antagonists could improve cognitive function, under conditions of pre-existing impairment due to overactive serotonergic inhibition, as is thought to occur in Alzheimer's disease. PMID- 11006721 TI - Is the context specificity of latent inhibition a sufficient explanation of learned irrelevance? AB - In three experiments, rats were pre-exposed either to uncorrelated presentations of a light and sucrose pellets (group CS/US) or to equivalent presentations of the light and pellets in separate sessions (control). In Experiment 1, subsequent conditioning to the light proceeded more slowly in group CS/US than in the control group, whether this conditioning was excitatory, with the light signalling the delivery of pellets, or inhibitory, with the light signalling their absence. Bonardi and Hall (1996) have argued that this learned irrelevance effect may be reducible to latent inhibition, which would be stronger in group CS/US because they are both pre-exposed and conditioned to the CS in the presence of traces of previous USs occurring in the same session. This analysis implies that group CS/US should have conditioned more rapidly to the CS than controls on the first trial of each session in Experiment 1, but this did not happen. It also implies that the learned irrelevance effect should be reversed if conditioning trials are given at a rate of one per day. Experiments 2 and 3 found no support for this prediction. We conclude that learned irrelevance effects cannot always be reduced to latent inhibition. PMID- 11006722 TI - Common coding in symbolic matching tasks with humans: training with a common consequence or antecedent. AB - The present experiments explored the importance of training with a common antecedent or consequence for common coding in symbolic matching-to-sample tasks with human subjects using reversal and transfer tests. Experiment 1 assessed common coding following training in many-to-one (MTO) and one-to-many (OTM) symbolic matching-to-sample tasks by comparing learning of partial and total reversals of these contingencies. Experiment 2 assessed common coding in MTO and OTM tasks by comparing performances in transfer tests when the common coding relations in the transfer phase were either consistent or inconsistent with those present during initial training. Evidence for common coding in the MTO and OTM discriminations was obtained in both experiments, although the amount of common coding was greater in the MTO discrimination in Experiment 1. These data are discussed in terms of associative mediational approaches to common coding phenomena. It is suggested that a basic requirement of an associative theory is that common coding should be more likely to occur when the stimuli are trained with a common consequence than when they are trained with a common antecedent, and that this requirement has been at least partly satisfied in the present studies. PMID- 11006723 TI - [Capital punishment]. PMID- 11006724 TI - [Challenges in forensic psychiatry]. PMID- 11006725 TI - [A votre sante!]. PMID- 11006726 TI - [Referral to a specialist]. PMID- 11006727 TI - [Should optometrists be able to refer patients to ophthalmologists?]. AB - BACKGROUND: Today optometrists in Norway can refer patients directly to ophthalmologists. Our article evaluates the quality of this practice. MATERIALS AND METHODS: 198 referrals from optometrists and 208 from general practitioners were evaluated. RESULTS: We found the referrals to be relevant to the ophthalmologist for nearly 95% in both groups. INTERPRETATION: The existing practice in which optometrists can refer their patients directly to ophthalmologists should be continued. PMID- 11006728 TI - [Hidden motor dysfunctions in children with epilepsy]. AB - BACKGROUND: At the National Center for Epilepsy, many years of clinical experience with children with difficult-to treat epilepsy has left the impression that many of these children may have motor difficulties in addition to their seizures. The aim of this prospective, uncontrolled study was to investigate if this impression could be confirmed. MATERIAL AND METHODS: In 41 children with "epilepsy only", i.e. without mental retardation or cerebral palsy, their motor performance was evaluated by means of the ABC test (Movement Assessment Battery for Children). RESULTS: More than half of the children had test results consistent with motor difficulties. The motor problems were most pronounced among those with MRI-detectable brain lesions and symptomatic localisation-related epilepsy. Gender, type and number of antiepileptic drugs or seizure frequency did not influence the test results. INTERPRETATION: Hidden motor dysfunctions, which may be caused by the aetiology of the epilepsy, the epilepsy itself, or the epilepsy treatment may easily be overlooked and may represent an extra burden for the children both at school and in their leisure time. We recommend that motor function should be tested in children with severe epilepsy as part of a comprehensive multiprofessional evaluation. Those with poor motor performance should be offered sensory motor stimulation. PMID- 11006729 TI - [Exposure therapy for anxiety disorders in a psychiatric ward]. AB - Exposure treatment is efficient as a treatment for some anxiety disorders. Briefly described, it consists in letting the patient expose himself to an anxiety-inducing situation without running away from it. In the author's experience, this mode of treatment is too seldom used. This article exemplifies conditions in which this form of therapy can be useful. The method is easily learnt by general practitioners. The article describes the treatment of five patients with these psychiatric diagnoses: agoraphobia, social phobia, compulsive disorder, and post-traumatic stress disorder. PMID- 11006730 TI - [Torsion of the Fallopian tube causing acute abdominal pain in a young virgin girl]. AB - BACKGROUND: Acute abdominal pain in female patients is a common condition often involving a gynaecological examination in order to establish an aetiological diagnosis. MATERIAL: The case of a 14-year-old girl with acute abdominal pain caused by isolated torsion of the right tube is presented. The literature is reviewed according to the discussion on whether an undiseased tube can undergo torsion. Theories concerning the pathogenesis of torsion and the premises for accepting the fallopian tube as previously undiseased, are presented. RESULTS AND INTERPRETATION: On the background of these premises we conclude that our patient might be the eighth case referred in the literature. PMID- 11006731 TI - [Forensic psychiatric risk assessment is ethically defensible. The dangerousness and risk of relapse are possible to judge with more precision than random assessment]. PMID- 11006732 TI - [Water channels of the cell--aquaporins]. AB - BACKGROUND: Several mechanisms in the body regulate the water transport within the cells up and down during physiological and pathological conditions. The discovery of aquaporins, water channel proteins, has brought more insight into and understanding of how water crosses plasma membranes within cells in body tissue. MATERIAL AND METHODS: The functions of aquaporins are not fully understood; this paper summarise current knowledge of various aquaporins. RESULTS: Known aquaporins are discussed in relation to some physiological, pathological and clinical conditions. INTERPRETATION: In the future, measurements of aquaporin concentrations in urine and possibly in other samples will give us a broader picture of water exchange and a better understanding of the mechanisms underlying water production in clinical states like renal and heart failure and liver insufficiency. Moreover, by manipulating water channel proteins by compounds developed for this purpose, one may have a tool for treating some clinical disorders in which water depletion or water overload is an important factor. PMID- 11006733 TI - [Drug therapy of patients with emotionally unstable personality]. AB - BACKGROUND: Most patients with borderline personality disorders are treated with psychotropic medication. There are, however, no clear clinical guidelines concerning optimal treatment of the disorder. MATERIAL AND METHODS: This article reviews the data on the efficacy of treatment with different classes of psychopharmacological drugs. The emphasis is on clinical trials published after 1981, as most placebo-controlled double-blind studies have been done in the last two decades. Furthermore, studies conducted before 1980 are more difficult to interpret because of changes in clinical diagnostic criteria. RESULTS: Neuroleptics have been studied most extensively followed by antidepressants. Neuroleptics have a modest, but broad therapeutic effect on symptoms in all domains. Doses are lower than those used for treating schizophrenia. Antidepressants have a more inconsistent effect. Tricyclics have been the least successful, whereas irreversible MAO inhibitors and selective serotonin reuptake inhibitors (SSRIs) have been effective in treating mood symptoms and impulsivity. Lithium has a possible effect in diminishing anger and suicidal symptoms. INTERPRETATION: As there is no "drug of choice" for the treatment of borderline personality disorder, a more rational clinical approach might be to treat different symptom clusters (cognitive/schizotypal, affective, impulsive) rather than the disorder itself. Finally, some practical guidelines for drug treatment of the disorder are proposed. PMID- 11006734 TI - [Relapse as quality indicator in psychiatric treatment]. AB - BACKGROUND: Efficiency and productivity expressed by length of hospitalisation, number of admissions, treated patients per time unit, and cost in NOK are incomplete measures for quality in a medical context. These measures should be supplemented by measures of results based on the course of illness for specific diagnostic categories. Few psychiatric institutions, including outpatient units, record the course of illness on a regular basis. MATERIAL AND METHODS: A review of the literature shows that rough measures of results, for instance readmission rates, often form the basis for evaluation of course/treatment results. The article gives examples of measures like readmission rates and more scientific measures such as Positive and negative syndrome scale (PANSS) used to measure results in a Norwegian psychiatric hospital. RESULTS: Of all first time admitted patients to Rogaland Psychiatric Hospital in 1985 and 1990, 40% of patients diagnosed with schizophrenia were readmitted after one year. For first time admitted patients from 1993 and 1994, the remission rate was 56% measured by PANSS. INTERPRETATION: The results of the treatment of first time admitted patients with a schizophrenia diagnosis were poorer than what is suggested possible through studies of optimal treatment, where the remission percentage is estimated to be 80% after one year. Rough measures of results, like readmission, do not seem to differ considerably from the results measured by PANSS. Norwegian psychiatric institutes are recommended to monitor the quality of treatment through a systematic recording of relapse among patients with a first time schizophrenia diagnosis. PMID- 11006735 TI - [Driving under the influence of medication and various substances other than alcohol]. PMID- 11006736 TI - [Screening for psychiatric disorders among prison inmates]. AB - BACKGROUND: Prison inmates have high frequencies of psychiatric disorders. Most prisons have little health-personnel resources, and methods to help focus resources towards those with serious health-care needs would be useful. The Global Symptom Index (GSI) of the Symptom Check List-90 (SCL-90) has performed well in other populations. MATERIAL AND METHODS: Prisoners at Kristiansand County Prison, Norway were asked to participate in the study, and 187 of 206 (91%) consented. All filled in the SCL-90 during the first four days of incarceration, and were examined clinically by a psychiatrist. RESULTS: Clinical examination revealed 40 persons with psychiatric disorder. Of these, 37 had a GSI score > or = 1. There were three false negatives and two false positives. Based on various cut-off levels for the GSI, we found a GSI cut-off value at 1.5 to perform best with sensitivity = 0.78, specificity = 0.87, and Number Needed to Diagnose = 1.55. INTERPRETATION: SCL-90 performs well as a screening instrument for psychiatric disorders among prison inmates. PMID- 11006737 TI - [Freedom as a medical problem--forensic psychiatry in a philosophical perspective]. PMID- 11006738 TI - [Preventive detention or custody--reflections over the new legal reactions]. PMID- 11006739 TI - [Serving a sentence on medical basis]. PMID- 11006740 TI - [Telemedicine--organizational consequences more than just talk?]. AB - In the telemedicine community, the possibility of organisational consequences of the technology have been mentioned frequently. But little has been published on the subject. We may speak of five categories of application of the new technology: clinical use, medical service functions, extramural applications, skills enhancement; and management and administration. Organisational consequences may be of four types: changes in distribution of tasks between levels of care and between organisations at the same levels, new division of work and change in skills; groups of individuals who are not co-located may co-operate in problem-solving, and the technology may become a tool of co-ordination for organisations. Empirical studies of the impact on organisation are few, but a Norwegian study confirmed that telemedicine already has such impact on the health service. PMID- 11006741 TI - [Telepathology at the Norwegian Radium Hospital]. AB - BACKGROUND: The article gives an overview of the telepathology activity at the Norwegian Radium Hospital from the service was launched in 1994 and up until today. We show the development during these years and discuss telepathology in general terms. We also discuss those aspects that determine how well a telepathology service functions. MATERIAL AND METHODS: 74 frozen section slides were diagnosed by two different telepathology systems. One of these systems was used for examining its appropriateness as a tool for second opinion in pathology. A new Internet-based system was developed that provided additional functionality. RESULTS: A telepathology system with a digital camera outperforms one with an analog camera with respect to diagnostic accuracy. INTERPRETATION: Image quality determines the precision of a telepathology service. Telepathology is a feasible tool for second opinion in pathology. PMID- 11006742 TI - [Telemedicine as means to recruit physicians to the districts]. PMID- 11006743 TI - [Stigma and stigmatization in psychiatry--a field which requires achievements?]. PMID- 11006744 TI - [European survey at the Center of Child and Adolescent Psychiatry]. PMID- 11006745 TI - [Hemovigilance--a new name or an old, unfulfilled need]. PMID- 11006746 TI - [Ovarian cancer screening of high risk groups]. PMID- 11006747 TI - [Unnuanced on cancer risk using snuff]. PMID- 11006748 TI - [Cardiologic procedures at a smaller Norwegian hospital--educational potential for assistant residents]. PMID- 11006749 TI - [New diagnostic criteria for diabetes mellitus]. PMID- 11006750 TI - [About the diagnostics of obturator hernia]. PMID- 11006751 TI - [Practice from Germany not sanctioned as specialist education in Norway]. PMID- 11006752 TI - [Latin in Norway and the Nordic countries]. PMID- 11006754 TI - Ecosystem evasion and health. PMID- 11006753 TI - [Pain relief and euthanasia]. PMID- 11006755 TI - A step toward putting a genie back in its bottle. PMID- 11006756 TI - Research ethics and a patient in her 70s. PMID- 11006757 TI - Jehovah's Witnesses and artificial blood. PMID- 11006758 TI - Comparing international infant mortality rates. PMID- 11006759 TI - The last trial of a Nazi doctor. PMID- 11006760 TI - Prevalence and predictors of human papillomavirus infection in women in Ontario, Canada. Survey of HPV in Ontario Women (SHOW) Group. AB - BACKGROUND: Human papillomavirus (HPV) is thought to be the primary cause of cervical intraepithelial neoplasia and cervical cancer. We determined the age specific prevalence of HPV infection and its risk factors in Ontario women. METHODS: We obtained 2 cervical specimens from randomly selected women (in 5-year age categories, from 15 to 49 years) who were being seen in 32 family practices for cytologic screening. The specimens were tested for carcinogenic HPV by the hybrid capture II assay (Digene Corp., Silver Spring, Md.) and by polymerase chain reaction (PCR) and genotyping. RESULTS: Of 1004 women eligible to participate, samples were obtained from 955 (95.1%). The prevalence of HPV (as determined by the hybrid capture II method) was highest, at 24.0% (95% confidence interval [CI] 16.5% to 31.5%), among women 20 to 24 years of age and was progressively lower in older age groups, reaching 3.4% (95% CI 0.1% to 6.7%) in women 45 to 49 years old. The prevalence of HPV (any type) as determined by PCR showed a similar pattern but was significantly higher (p = 0.01) among women 45 to 49 years old than among those 40 to 44 years old (13.0% [95% CI 6.4% to 19.6%] v. 3.3% [95% CI 0.1% to 6.5%]). Risk factors for positivity with the hybrid capture II method were never-married status, divorced or separated status, more than 3 lifetime partners, more than 1 partner in the preceding year, cigarette smoking and current use of oral contraceptives. The presence of squamous intraepithelial lesions on cytologic examination was strongly associated with positivity with the hybrid capture II assay (odds ratio 96.0, 95% CI 22.3 to 413.4; p < 0.01). INTERPRETATION: The highest prevalence of HPV was 24.0%, in women 20 to 24 years old. Risk factors supported a sexual mode of transmission, and there was a strong association between HPV and abnormal cervical cytologic results. PMID- 11006761 TI - Comparison of self-collected vaginal, vulvar and urine samples with physician collected cervical samples for human papillomavirus testing to detect high-grade squamous intraepithelial lesions. AB - BACKGROUND: Certain types of human papillomavirus (HPV) in cervical samples are strongly associated with squamous intraepithelial lesions (SIL) and invasive cervical carcinoma. We determined and compared the test characteristics of testing for HPV with samples obtained by patients and with samples obtained by their physicians. METHODS: In a consecutive series of women referred to a colposcopy clinic at a teaching hospital because of abnormalities on cervical cytologic screening, 200 agreed to collect vulvar, vaginal and urine samples for HPV testing. The physician then collected cervical samples for HPV testing, and colposcopy, with biopsy as indicated, was performed. Presence of HPV was evaluated using the hybrid capture II assay (Digene Corp., Silver Spring, Md.) with a probe cocktail for 13 carcinogenic types. Cervical specimens were also tested for HPV by polymerase chain reaction and hybridization with type-specific probes. Cervical smears for cytologic examination were obtained from all women. RESULTS: High-grade lesions (high-grade squamous intraepithelial lesions [HSIL], equivalent to cervical intraepithelial neoplasia [CIN] grade 2 or 3, and adenocarcinoma) were found in 58 (29.0%) of the 200 women. Carcinogenic types of HPV were detected in the self-collected vaginal samples of 50 (86.2%) of these 58 women, in the self-collected vulvar samples of 36 (62.1%) and in the self collected urine samples of 26 (44.8%). Carcinogenic types of HPV were detected in the cervical samples collected by physicians for 57 (98.3%) of these 58 women. The remaining 142 women (71.0%) had normal findings or low-grade squamous intraepithelial lesions (LSIL, CIN grade 1). Test results were negative or noncarcinogenic types of HPV were detected in the self-collected vaginal samples of 76 (53.5%) of these 142 women, in the self-collected vulvar samples of 89 (62.7%) and in the self-collected urine samples of 99 (69.7%). The sensitivity for self-collected samples ranged from 44.8% to 86.2%, and the specificity from 53.5% to 69.7%. For the samples collected by physicians, the sensitivity was 98.3% and the specificity 52.1%. The self-sampling methods were generally acceptable to the women: 98.4% of respondents (126/128) deemed urine sampling acceptable, 92.9% (118/127) found vulvar sampling acceptable, and 88.2% (112/127) found vaginal sampling acceptable. INTERPRETATION: Self-collection of samples for HPV testing was acceptable to women attending a colposcopy clinic for investigation of suspected cervical lesions and shows sufficient sensitivity to warrant further evaluation as a screening test for cervical cancer prevention programs. PMID- 11006762 TI - Evaluating the benefits of antimicrobial prophylaxis to prevent urinary tract infections in children: a systematic review. AB - BACKGROUND: The recurrence rate for urinary tract infections in children is estimated at between 30% and 40%. The use of low doses of antibiotics as prophylaxis for recurrent urinary tract infections is common clinical practice. However, prolonged antimicrobial therapy has the potential to contribute to problems of bacterial resistance and antimicrobial side effects. The aim of this review was to systematically examine the available evidence for the effectiveness of this intervention. METHODS: We conducted a literature search of 3 electronic databases for the period 1966 to 1999. We also searched bibliographies from conference proceedings and contacted content experts to ensure completeness of our database. Each trial was evaluated on the basis of the following inclusion criteria: target population (children), intervention (antibiotic v. no antibiotic), outcome (number of urinary tract infections) and study design (randomized controlled trial). Quality was assessed for the studies that met these criteria. RESULTS: Most of the studies identified were case series and cohort studies. Only 6 randomized trials fulfilled the inclusion criteria. All were of low quality (median 2, range 0 to 2 [maximum quality score 5]). Three trials dealt with children who had anatomically normal urinary tracts, and three included children with neurogenic bladder. The rate of infections for patients with normal urinary tracts ranged from 0 to 4.0 per 10 patient-years for the treatment groups and from 4.0 to 16.7 for the control groups. The recurrence rates for patients with neurogenic bladders in 2 trials were 2.9 and 17.1 per 10 patient-years for the treatment groups and 1.5 and 33.0 for the control groups. INTERPRETATION: The available evidence for using antimicrobial prophylaxis to prevent urinary tract infection in children with normal urinary tracts or neurogenic bladder is of low quality. This suggests that the magnitude of any benefit should at best be questioned. The surprising lack of data for children with reflux is of concern. Well-designed trials are needed to optimize the use of antimicrobials in children with recurrent urinary tract infection. PMID- 11006763 TI - Environment and health: an overview. PMID- 11006764 TI - Screening for cervical cancer: should we test for infection with high-risk HPV? PMID- 11006765 TI - Colorectal cancer screening: now is the time. PMID- 11006766 TI - Population-based fecal occult blood screening for colon cancer: will the benefits outweigh the harm? PMID- 11006767 TI - Environment and health: 1. Population, consumption and human health. PMID- 11006769 TI - Postoperative tetanus: a case report. PMID- 11006768 TI - Type 2 diabetes mellitus in Canada's first nations: status of an epidemic in progress. AB - This review provides a status report on the epidemic of type 2 diabetes mellitus that is affecting many of Canada's First Nations. We focus on the published literature, especially reports published in the past 2 decades, and incorporate data from the Aboriginal Peoples Survey and the First Nations and Inuit Regional Health Survey. We look at the extent and magnitude of the problem, the causes and risk factors, primary prevention and screening, clinical care and education, and cultural concepts and traditional knowledge. The epidemic of type 2 diabetes is still on the upswing, with a trend toward earlier age at onset. Genetic environmental interactions are the likely cause. Scattered intervention projects have been implemented and evaluated, and some show promise. The current health and social repercussions of the disease are considerable, and the long-term outlook remains guarded. A national Aboriginal diabetes strategy is urgently needed. PMID- 11006770 TI - FDA considering restricted access to "abortion pill". PMID- 11006771 TI - More net-savvy MDs surfing the Web. PMID- 11006772 TI - MDs get jail terms, fines as new police squad targets health fraud. PMID- 11006773 TI - RNs seek broader prescribing powers in quest for more autonomy. PMID- 11006774 TI - COX-2 inhibitors and renal function in elderly people. PMID- 11006775 TI - Utility of the clinical examination for carpal tunnel syndrome. PMID- 11006776 TI - Anthrax: of bison and bioterrorism. PMID- 11006777 TI - Benchmarking: a prescription for healthcare. PMID- 11006778 TI - Assurance services and healthcare. PMID- 11006779 TI - The nursing shortage. A difficult conversation. PMID- 11006780 TI - Is now the time to design new care delivery models? PMID- 11006781 TI - Linking practice and education. PMID- 11006782 TI - Nursing administrative practicum. Enhancing communication between staff nurses and nursing leadership. PMID- 11006783 TI - Organizational trust and empowerment in restructured healthcare settings. Effects on staff nurse commitment. AB - In today's dramatically restructured healthcare work environments, organizational trust is an increasingly important element in determining employee performance and commitment to the organization. The authors used Kanter's model of workplace empowerment to examine the effects of organizational trust and empowerment on two types of organizational commitment. A predictive, nonexperimental design was used to test Kanter's theory in a random sample of 412 Canadian staff nurses. Empowered nurses reported higher levels of organizational trust, which in turn resulted in higher levels of affective commitment. However, empowerment did not predict continuance commitment--that is, commitment to stay in the organization based on perceived lack of other job opportunities. Because past research has linked affective commitment to employee productivity, these results suggest that fostering environments that enhance perceptions of empowerment and organizational trust will have positive effects on organizational members and increase organizational effectiveness. PMID- 11006784 TI - Building community in the healthcare workplace. Part 2, Envisioning the reality. AB - Squeezed between mounting budget pressures and staffing demands, healthcare managers have little room to maneuver as they are buffeted by incessant new strategies. So what is the constant that they can use to keep a steady course through all the chaos? We suggest that the overarching constant is "community in the workplace." Although community building may have both short- and long-term benefits, it is not a strategy but a career-long philosophy, structure, and approach to help managers tie together all the facets of their daily work into a more meaningful and satisfying purpose. This article, part 2 in a 4-part series, explores the principles and characteristics of community and provides material for further study and discussion. Four conceptual cornerstones--philosophy, influence, culture, and inner transformation--are considered in relation to the community approach. Part 1 (April) described the negative dynamics that obstruct sustainable change and reasons for moving to community at work. Part 3 (October) will describe how to build community and will give options for implementing change in complex adaptive systems. The final article (November) presents case studies and discusses how community can be built in a unionized environment. PMID- 11006785 TI - Skills needed for promotion in the nursing profession. AB - OBJECTIVE: This research was designed to identify skills (interpersonal versus technical) important to promotion within the nursing profession. Also studied was the difference between actual and perceived importance of the skills. BACKGROUND: For many technical and academic professions, career advancement is attained through promotions. These promotions often take one further away from the front line; thus, the technical skills so important in one's early career become less important. Interpersonal skills, on the other hand, increase in importance when one is supervising other professionals. Though research has described this phenomenon, it is not known exactly where the shift in importance occurs or what the precise nature of the shift may be. Further research in this area is particularly important to management development programs in technical, scientific professions. METHODS: A policy-capturing approach was used to identify the skills important to managerial advancement in the nursing profession. Two hundred nineteen nurse administrators from a large Southeastern United States hospital rated hypothetical candidates for managerial positions. At the lowest level management position studied, a greater number of technical skills significantly influenced promotion decisions than at middle management. Significant differences were found between decision-makers' perceptions of skill importance and the skills' actual importance to promotion decisions. RESULTS: The present research indicate that a greater number of technical skills are important for promotion to CN3 than for promotion to ANM. The results also suggest decision makers are not always 100% accurate in their insight regarding the specific factors influencing their personnel decisions. PMID- 11006786 TI - Development of a nursing management practice atlas. Part 2, Variation in use of nursing and financial resources. AB - Developing mechanisms for making benchmark comparisons among hospital organization is a challenge that has been embraced by nurse executives. A methodologic approach for ensuring data congruency when using available secondary data bases for making benchmark comparisons was detailed in part one (July/August) of this two-part series. This second article analyzes nursing management data using a set of nursing and financial resource variables identified by senior nurse executives of the hospital sites involved in this study. PMID- 11006788 TI - Treatment of diffuse parenchymal lung disease. Part 2. PMID- 11006789 TI - Tinzaparin is newest entry into low-molecular-weight heparin market. PMID- 11006790 TI - Argatroban approved for heparin-induced thrombocytopenia. PMID- 11006791 TI - Iowa Medicaid to begin paying pharmacists in case management project. PMID- 11006792 TI - Better outcomes, higher costs associated with patient-centered hospital units. PMID- 11006787 TI - The diagnosis, assessment and treatment of diffuse parenchymal lung disease in adults. Introduction. PMID- 11006793 TI - Analysis finds modest effect for antimicrobials in acute otitis media. PMID- 11006794 TI - Pharmacy technician's role in an ambulatory care infusion clinic. PMID- 11006796 TI - Oral serotonin type 3-receptor antagonists for prevention of chemotherapy-induced emesis. AB - The theoretical basis for and clinical experience with using oral serotonin type 3 (5-HT3)-receptor antagonists for preventing chemotherapy-induced emesis are discussed. Evidence supports the idea that antineoplastic drugs and irradiation can initiate emesis by releasing serotonin from enterochromaffin cells in the gut mucosa, which activates peripheral vagal afferent nerves. In view of the GI site of serotonin release and vagal afferent activation, the proximity of neuronal 5 HT3 receptors, and the pharmacologic properties of 5-HT3-receptor antagonists, the oral use of these agents is rational. Oral granisetron 2 mg once daily or 1 mg twice daily has been evaluated in more than 4500 patients receiving highly or moderately emetogenic chemotherapy. Rates of total control of emesis ranged from 44% to 60%, and complete-response rates ranged from 70% to 94%. Oral ondansetron 8 mg three times daily has proven effective in patients receiving antineoplastics with moderate or moderately high emetogenic potential. Two double-blind studies demonstrated the efficacy of a single 24-mg oral dose of ondansetron administered approximately 30 minutes before cisplatin-based chemotherapy. Patients randomized to oral ondansetron had higher total-control and complete-response rates than patients receiving intravenous granisetron or ondansetron. Oral dolasetron 100 or 200 mg once daily also prevented emesis. Oral administration of 5-HT3-receptor antagonists for the prevention of acute emesis associated with chemotherapy is rational and appears to be effective. PMID- 11006795 TI - Raloxifene hydrochloride. AB - The pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and therapeutic role of raloxifene hydrochloride are reviewed. Raloxifene is a selective estrogen-receptor modulator (SERM) that has been approved for use in the prevention and treatment of osteoporosis in postmenopausal women. A SERM interacts with estrogen receptors, functioning as an agonist in some tissues and an antagonist in other tissues. Because of their unique pharmacologic properties, these agents can achieve the desired effects of estrogen without the possible stimulatory effects on the breasts or uterus. Raloxifene is rapidly absorbed from the gastrointestinal tract and undergoes extensive first-pass glucuronidation. Approximately 60% of a dose is absorbed; however, absolute bioavailability is only 2%. The volume of distribution is 2348 L/kg for a single oral dose of 30-150 mg, and the elimination half-life averages 32.5 hours. In clinical trials in postmenopausal women, raloxifene had an estrogen-like effect on bone turnover and increased bone mineral density. It reduced the risk of fractures in women with osteoporosis. Raloxifene also seemed to reduce the risk of breast cancer and positively influenced blood lipid markers of cardiovascular disease. Raloxifene is generally well tolerated; the most common adverse effects are hot flashes and leg cramps. A serious adverse effect is venous thromboembolism. The recommended dosage is 60 mg/day orally without regard to meals. Ultimately, it will be information on cardiovascular or breast cancer benefits that will determine the future role of raloxifene. Raloxifene is an alternative to traditional hormone replacement therapy for the prevention and treatment of osteoporosis in selected postmenopausal women. More study is needed to verify possible benefits related to heart disease and breast cancer. PMID- 11006797 TI - Comparison of three a priori methods and one empirical method in predicting lithium dosage requirements. AB - The precision and bias of three a priori methods and an empirical method for predicting lithium dosage requirements were studied. Data on serum lithium concentrations were collected from inpatient medical records at a state psychiatric hospital. Predicted daily lithium doses were calculated by using a priori methods proposed by Zetin et al., Jermain et al. and Pepin et al., and an empirical method and compared with the patients' actual dosages. Similar comparisons were made with respect to serum lithium concentrations at steady state. Absolute mean error and mean error were calculated to assess the precision and bias of each a priori method. The records of 47 patients were used in the study. Average mean error for dosage predictions was -130.41, -187.69, 170.80, and -357.23 mg/day for the Jermain, Pepin, Zetin, and empirical methods, respectively. Average mean error for serum lithium concentration predictions was 0.11, -0.09, and 0.37 meq/L for the Jermain, Pepin, and empirical methods, respectively. The Jermain and empirical methods significantly overpredicted concentration and underpredicted dosage. The Zetin method overpredicted dosage. The Pepin method underpredicted dosage but not concentration. The average difference in dosage error among the methods was only 73.3 mg/day. Three a priori dosing methods were similar to an empirical method in their ability to predict lithium dosages. All methods were biased. Although all a priori methods were more precise than the empirical method, the clinical significance is unclear. PMID- 11006798 TI - Conversion of patients from simvastatin to lovastatin in an outpatient pharmacy clinic. PMID- 11006799 TI - Variations in finishing time of 24-hour chemotherapy infusions. PMID- 11006800 TI - Thinking out of the box. PMID- 11006801 TI - Costs and bacterial susceptibility after therapeutic interchange of fluoroquinolones. PMID- 11006802 TI - Cost of cerivastatin in cost-effectiveness study. PMID- 11006803 TI - Muscle relaxants and renal failure. PMID- 11006804 TI - Under renovation: incorporating change. PMID- 11006805 TI - Evolving narratives in the course of retirement: a longitudinal study. AB - This paper presents results from the second phase of a longitudinal study of retirement. Data were gathered through interviews with 29 participants (65 to 66 years of age) who had previously been interviewed when they were 63 to 64 years of age and still working. Data were analyzed by characterizing each subject's narrative about retirement in terms of its narrative slope--progressive, stability, or regressive. These current narratives also were compared with the earlier narratives these participants told. It was found that, while anticipatory narratives may predispose persons toward action, they are not so much a fixed "script for action" as an orientation to act within circumstances. While participants' narratives anticipating retirement often turned out as expected, they were sometimes reshaped as a consequence of personal action, external events, and unexpected experiences within new retirement occupations. The study suggests that, while narratives play a role in shaping the direction of persons lives, they also interweave with and change directions as a result of ongoing life events and experiences. PMID- 11006806 TI - The predictive power of narrative data in occupational therapy evaluation. AB - OBJECTIVE: This study examined whether adding the Canadian Occupational Performance Measure (COPM) to existing occupational therapy evaluation measures used in a subacute skilled nursing facility unit enhanced the accuracy of therapists' predictions of the functional status of clients at discharge. METHOD: This study utilized a prospective comparison design. Two independent predictive variables were developed using the standard Functional Independent Measure (FIM), and an enhanced FIM that included narrative information from the Canadian Occupational Performance Measure (FIM/COPM). These variables were subsequently compared with the actual FIM discharge (DFIM) scores for 31 clients. The primary author (D.S.) gathered data from chart review and conducted the statistical analysis. The data were analyzed using descriptive correlations (Pearson r) and comparison statistics (Wilcoxon signed rank test). RESULTS: Comparison statistics (Wilcoxon signed rank test) revealed a statistically significant difference between the standard FIM predictive score and the discharge FIM score. No statistically significant difference was found between the FIM/COPM predictive score and the discharge FIM score. These findings suggest that predictive scores based solely on information attained from the standard FIM resulted in less accuracy in outcome predictions. Correlational analyses further supported these conclusions. CONCLUSION: The findings support the study hypothesis that use of the COPM in combination with the FIM enhances accuracy in prediction of outcomes for rehabilitative services for persons in adult physical disabilities settings. PMID- 11006807 TI - The effects of hands-on occupation versus demonstration on children's recall memory. AB - OBJECTIVE: Hands-on learning has been an important aspect of the profession of occupational therapy since its founding. The purpose of this study was to determine whether children engaged in hands-on learning would be able to recall more of the steps and more of the correct order of the steps of an occupation than children engaged in a demonstration teaching method. METHOD: After being randomly assigned, 73 healthy third-graders (42 girls and 31 boys) either participated in making a model of a volcano or observed the making of a model of a volcano. Following task completion, both groups were asked to recall and state as many of the 41 syntactical units as possible in their proper order. The children's responses were audiotaped and scored in a blind fashion according to predetermined criteria. Interrater reliability was excellent. RESULTS: A t-test revealed a significant difference between conditions in terms of free recall scores, with children in the hands-on condition having a greater recall score, t (71) = 2.63, p < .005. The effect size d equaled .62. A MannWhitney U Test revealed no significant difference between conditions in terms of remembering the steps in proper order (the lack of a significant difference may have been due to a problem of measurement). CONCLUSION: The results of this study demonstrated that participants were able to recall more information when engaged in a hands-on teaching method as compared with a demonstration method. It is suggested that the learning advantages of hands-on occupation are related to the enhanced sensory/perceptual experiences and the feelings of success that are characteristic of hands-on learning, as opposed to passive forms of learning. PMID- 11006808 TI - Reliability and validity of a parent questionnaire on childhood motor skills. AB - OBJECTIVE: As the consequences of clumsiness in children become better understood, the need for valid measurement tools is apparent. Parent report has the potential for providing historical knowledge of the child's motor skills, as well as perceptions of their children's motor difficulties. The objective was to develop a parent questionnaire to identify motor difficulties in children. METHOD: A sample of 306 children participated in the development of a 17-item parent questionnaire, called the Developmental Coordination Disorder Questionnaire (DCDQ). Internal consistency, concurrent and construct validity were examined. RESULTS: The DCDQ proved capable of distinguishing children who had motor problems (as measured by standardized tests) from children without motor problems. Correlations with standardized tests were significant. Two other studies confirmed the construct validity of the DCDQ. Factor analysis revealed four distinct factors, useful in defining the nature of the difficulties. CONCLUSION: The DCDQ is a succinct and useful measure for use by occupational therapists. PMID- 11006809 TI - The evaluation of sensory processing: a validity study using contrasting groups. AB - The purpose of this study was to examine the validity of a sensory history questionnaire using contrasting groups. Specifically, the goal was to identify items on the Evaluation of Sensory Processing that differentiate between parent ratings of children with sensory integrative dysfunction and parent ratings of typically developing children. Data were collected from 30 parents of children with sensory integrative dysfunction and 59 parents of typically developing children. Items were analyzed using a Wilcoxon signed rank test to detect the difference between the parent ratings for the 30 children with sensory integrative dysfunction and 30 of the typically developing children who were matched to the dysfunctional children on age, ethnicity, gender, geographic location, and socioeconomic status. Eighty-four of the 200 items significantly (p < .05) distinguished between parents' ratings of children with and without sensory integrative dysfunction. PMID- 11006810 TI - Relationship between choice and quality of life among residents in long-term-care facilities. AB - OBJECTIVE: This study tested the association between perceptions of personal control and quality of life among older persons. METHOD: Two self-report instruments. The Quality of Life Rating (QOLR) and the Duncan Choice Index (DCI), were administered to 21 residents in a long-term-care facility. The DCI was developed for this study to measure the amount of choice available in 29 self care and leisure activities. RESULTS: A significant positive correlation (r = .54; p = .01) between the amount of choice residents perceive they have and their quality of life was found. The DCI was shown to be reliable with preliminary evidence of construct validity. CONCLUSION: Enhancing personal control in everyday life may be associated with improved quality of life. Occupational therapy strategies to empower residents through increasing choice and control include increasing community in the facility emphasizing personal responsibility, and enabling choices in everyday tasks. PMID- 11006811 TI - Role perceptions of occupational therapists providing support and education for caregivers of persons with dementia. AB - OBJECTIVE: This pilot study explored occupational therapists' perceptions of their roles as interventionists providing education and support for caregivers of persons with dementia. The intervention was provided in caregivers' homes as part of a larger funded study. METHOD: Interviews were conducted with four occupational therapists to elicit their reflections on practice and their views on occupational therapy services on the basis of their experiences providing support and education for caregivers in the funded study. RESULTS: Key themes consisted of the contrasts between the therapists' roles in the study and their customary practices and the professional and personal impact of their role in the study. Their recommendations for occupational therapy services emphasized the need to (a) collaborate with patients, families, and other health care staff members to solve problems; (b) acknowledge others as experts; (c) include family perspectives; and (d) fully address the needs of patients and families in their home environments. CONCLUSION: Providing support and education for caregivers in the community can be a major transition for therapists accustomed to practicing in more traditional settings. Additional research is needed to explore the ways in which specific practice contexts influence delivery of occupational therapy services. PMID- 11006812 TI - The Joint Protection Behavior Assessment: a reliability study. AB - OBJECTIVE: Of the various measures developed for studying persons with rheumatoid arthritis, only one that focuses on joint protection has undergone extensive testing, the Joint Protection Behavior Assessment (JPBA). The purpose of the present study was to examine the interrater and intrarater reliability of the JPBA. METHOD: Six healthy participants performed the JPBA under three test conditions (uninformed, informed, completely guided joint protection behavior). The 18 test performances were videotaped and scored by nine independent raters. RESULTS: Analysis of these data showed that interrater reliability (intraclass correlation coefficient [ICC]) was .90 or higher, and intrarater reliability was .95 or higher (ICC). The correlation between the JPBA and its two shortened versions was .95 or higher (ICC). Internal consistency was also high, with a coefficient alpha of 0.95 for the complete JPBA. Kappa values showed that for most subtasks, there was fair to excellent agreement between raters and consistency of raters over time. CONCLUSION: Our data suggest that the complete JPBA has excellent clinimetric properties and that the shortened versions are adequate for clinical situations. Some improvements in the test manual suggested by the present study may further improve the measure. A repeat of this study under real-world circumstances would provide an estimate of JPBA reliability in clinical practice. PMID- 11006813 TI - Perceived risk as a constraint on occupational performance during hot and cold water pouring. AB - OBJECTIVE: Occupational therapists are interested in quality of movement under different environmental conditions. It has been shown that during reaching tasks, the physical aspects of the objects to be grasped can influence the quality of movement. This study investigated whether perceived risk (water temperature) affected the quality of movement during a pouring task. METHOD: In a counterbalanced, repeated measures design, 56 participants (M = 27.1 years, SD = 7.4 years) poured hot, then cold water to prepare hot and cold beverages. Dependent variables included movement time, displacement, peak velocity, percentage of movement time to peak velocity, and movement units. Data were analyzed with paired t tests. RESULTS: Participant performance in displacement and movement time was significantly different when considering the entire pouring task for both the hot and the cold conditions (ps < .05). In addition, significant differences were found in the discrete movement "sub-portions" of the pouring task (ps < .05). CONCLUSION: Perceived risk is an element of meaning that the occupational therapist can consider in providing the person with just enough challenge to facilitate successful performance. By varying the amount of perceived risk in the occupational form, the therapist can help the person experience and develop the range of movement strategies required by everyday occupations. Future research is needed to corroborate these findings in simple and more complex occupations as well as to examine perceived risk in special populations. PMID- 11006814 TI - Perspectives on faculty clinical practice: views of occupational therapy curriculum chairpersons and faculty members. AB - OBJECTIVE: This study investigated whether and how faculty clinical practice (FCP) was being implemented in occupational therapy professional education programs. METHOD: Chairpersons and faculty members from all accredited entry level occupational therapy curricula in the United States were asked to complete questionnaires about their involvement in FCP. The chairperson questionnaire consisted of primarily closed-ended questions that addressed the organization of faculty practice within their programs, whereas the faculty questionnaire was primarily open-ended and addressed faculty members' individual involvement in FCP as well as the perceived benefits and drawbacks. Responses were analyzed for 39 program chairpersons and 136 faculty members were analyzed. RESULTS: Twenty-five chairperson respondents reported that their educational program had FCP, and 44 faculty respondents indicated involvement in FCP during the past year. FCP was more prevalent in public-funded institutions and academic health centers. Faculty members primarily engaged in FCP in order to stay current, enhance teaching, and develop networks. They reported increased credibility with students as an important benefit of clinical practice. The primary reasons faculty members did not engage in clinical practice were insufficient time and institutional policies. CONCLUSION: Although many faculty members in occupational therapy education programs value clinical practice as a part of their educator role, it is necessary to negotiate responsibilities and rewards to prevent role overload and comply with institutional policies. PMID- 11006815 TI - Can we improve outcomes research by expanding research methods? AB - Therapeutic process occurs individually and in groups. The outcome of intervention, however, is demonstrated through changes in client performance in treatment sessions, that is, in individuals. Individual function, then, is both the target of intervention and the measure of effectiveness of occupational interventions. Reporting occupational therapy outcomes necessitates culling functional performances of individuals to describe the overall outcomes of an occupational therapy program (Johnson, 1996). In a summary report, a manager might combine outcomes from inpatient and outpatient services to depict the overall impact of occupational therapy services. For example, the manager might conclude that clients receiving occupational therapy services have an 85% rate of regaining independent living in the community. The challenge in reporting overall outcome is to find ways to demonstrate more clear links between individual treatment and function. Conclusions about groups of clients are often presented in the form of baseline and discharge comparisons that are easy to summarize. But relying on pre- and posttesting may deprive us of valuable information that can be gained by session-based analyses. Although it might appear cumbersome to examine individual session outcomes, collecting outcomes data is done informally by every therapist and client at the end of a treatment session. We suggest that this informal procedure become a formal part of outcomes research and a more routine part of occupational interventions. PMID- 11006816 TI - The use of nerve blocks in conjunction with occupational therapy for complex regional pain syndrome type I. PMID- 11006817 TI - Occupational therapy in early intervention: applying concepts from infant mental health. PMID- 11006818 TI - Test-retest reliability of the Purdue Pegboard Test. PMID- 11006819 TI - Teaching evidence-based practice. PMID- 11006820 TI - [Forensic medicine experiences with methadone substitution in the Geneva canton]. AB - Methadone treatment for heroin addiction has followed three distinct periods in Geneva, Switzerland. The first period (1970-1979) corresponds to the beginning of the heroin addiction epidemic. Treatment was restricted to detoxification and did not succeed in reducing fatal overdoses. During the second period (1980-1989), methadone maintenance program was favoured but access to this program was limited. This period has brought a decrease of illegal heroin consumption and criminality but not of fatal overdoses. Finally, during the third period (since 1990), legislation was changed to allow easier access to methadone maintenance program. As a consequence there was a significant drop in lethal heroin overdoses and in deaths attributed to HIV. PMID- 11006821 TI - [Alcohol stove as a source of CO poisoning in a camper]. AB - Although the number of deaths from fatal CO poisoning has strongly declined since the change from coal gas to natural gas, accidental and suicidal carbon monoxide intoxications still have to be expected. In motor vehicles the exhaust gases from the engine and a stationary heating are the major sources of intoxication. In closed campers and caravans the operation of gas and spirit cooking stoves etc. may also lead to an accumulation of carbon monoxide due to incomplete combustion. PMID- 11006822 TI - [Similar skin lesions in victim and perpetrator caused by a knife with a serrated blade]. AB - Reported in this paper is an attack of two adolescents on a man who was killed in the fight, with several kitchen knives being used, including two with grooved and wave-grooved blades. One of the offenders held the victim tight from behind and was injured by his attacking accomplice++. A grid mark on the left side of the victim's face and the left forearm of the second offender in the back supported the assumption of a knife with simple wave profile. Skin lesions of finer structure below the left ear and on the left forearm of the victim suggested involvement of a smaller kitchen knife with groove-milled wave profile. Offender victim position and course of offence were verified and confirmed by evaluation of these specific findings. PMID- 11006823 TI - [Fatal hunting injuries in North Rhine-Westphalia]. AB - The investigation covers 14 fatal hunting accidents which occurred in North Rhine Westphalia in the last 23 years, among them 13 firearm fatalities. All the accidents happened during the statutory hunting season reaching a peak in the main small game season. 86% of the persons causing the accidents and 73% of the victims were over 41 years and experienced hunters. 36% of the accidents were caused by the victims themselves. The fatal gunshot injuries were inflicted by rifle bullets in 7 cases (54%) and shotgun pellets in six cases (46%). No accidents were due to shotgun slugs or handgun ammunition. The main factors responsible for the fatalities were: covering the victim while swinging on the game (31%), failure to discharge the firearm while descending from the raised hide or transporting the weapon in vehicles (31%), mistaking the victim for game (15%), failure to discharge the weapon during a hunting break or while pressing down obstacles or careless handling of the weapon (23%). In no case was the accident caused by a defective weapon or ammunition. In conclusion aspects of preventing fatal hunting accidents are discussed. PMID- 11006824 TI - [Postmortem dog bites after sexually motivated homicide with multiple stab wounds -differential diagnostic aspects]. AB - The body of a 20-year-old woman that had undergone significant putrefaction was found lying on the bed of her apartment in a supine position. The legs were straddled and apart from a T-shirt, she was naked. The inner side of the right thigh showed a circumscribed 20 cm x 40 cm defect exposing the underlying muscles. The wound margin was ragged and superficial scratch-type abrasions were found in the vicinity of the injury margin. The investigating police officers classified the injuries to be caused postmortem by a crossbreed dog that was inside the apartment. No other signs of external force were found on the body at the death scene and due to the previous history of the deceased, a drug death was assumed. Autopsy revealed 13 vital stab wounds in the neck and aspiration of blood in the lungs as well as signs of manual strangulation. The observed coincidence of injuries due to postmortem animal depredation and stab wounds, although in different localization, can be regarded as a rare entity in the light of the literature. The morphological appearance of traumatic injuries due to other causes can be modified by the simultaneous presence of postmortem injuries caused by animals and difficulties can arise concerning the differential diagnosis especially under poor conditions (e.g. advanced putrefaction) at the death scene. PMID- 11006825 TI - [Distribution of morphine and morphine glucuronides in body tissue and fluids- postmortem findings in brief survival]. AB - An intoxication following administration of morphine, tramadol and atracurium in a suicide case is reported. The route of administration and the amount of the particular drug were known from the investigation of the death scene and the findings of the postmortem examination. Tramadol was present in the gastric contents as well as in blood, liver, kidney and brain samples, whereas the drug could not be detected in muscle. All body fluids and tissues investigated contained morphine as well as its 3- and 6-glucuronides with the exception of muscle tissue. The concentrations of morphine and its glucuronide metabolites were determined by LC/MS following solid phase extraction. Interestingly, the concentration of M6G in brain, liver and kidney were close to the concentration of M3G in the particular tissue. This phenomenon might be explained by a preferential hydrolysis of M3G or by a preferential formation of M6G postmortem. Measurement of morphine and M6G in femoral blood and cerebrospinal fluid may be a useful indicator in rapid deaths. PMID- 11006826 TI - [Significance of mitochondrial DNA for forensic stain analysis, identification and forensic lineage determination]. AB - DNA testing using conventional STR systems may produce insufficient results, if the genomic DNA in the specimen is either highly degraded or the available quantity is very small (e.g. skin particles, hair shafts or ancient bones). In some of these cases the examination of mitochondrial DNA, which is present in considerably larger copy numbers in the cytoplasm, is more successful than that of nuclear DNA. Identification of unknown corpses by conventional DNA typing sometimes remains doubtful, if only samples from presumably distant relatives or putative brothers or sisters are available for comparison. Since mitochondrial DNA is generally transmitted in maternal lineages, its sequence pattern can be directly compared with those of other individuals and, in case of the same maternal lineage, corresponding sequence chromatograms are to be expected. In connection with nuclear DNA typing methods certain sequence motives may furnish clues to ethnic groups. The report presents three cases illustrating the application possibilities of mtDNA typing in forensic practice. PMID- 11006827 TI - Dietary carbohydrates affect caecal fermentation and modify nitrogen excretion patterns in rats. I. Studies with protein-free diets. AB - In a two-factorial experiment on 96 young male rats, the effects of substituting 10% raw potato starch (PS), pectins (PEC), or cellulose (CEL) for corn starch (CS) were studied using an unsupplemented protein-free (PF) diet or a PF diet supplemented either with DL-methionine or urea. The pH and the short chain fatty acids (SCFA) content in caecal digesta, as well as caecal digesta and tissue weights were determined and used as the criteria of caecal fermentation intensity. Blood urea level, amount of N excreted via faeces and urine, DAPA content, and amino acid composition of faecal protein were analyzed as indices of protein metabolism. A 10-day adaptation period to the carbohydrates fed with the casein diet preceded the experimental period of feeding the respective carbohydrates with protein-free diets. Dietary carbohydrates significantly influenced total and individual SCFA content in caecal digesta, as well as other parameters related to the intensity of fermentation. Potato starch and pectins were more intensively fermented than cellulose. Supplementation of the PF diet with methionine and urea affected only caecal isobutyric and valeric acid content in a way dependent on the carbohydrates present in the diet. Carbohydrates significantly altered the routes of N excretion. Faecal excretion was increased by all carbohydrates studied compared to corn starch, pectins had the most marked effect. Urinary excretion was significantly increased by cellulose (as compared with the PEC and PS groups) and decreased by pectins as compared with all other groups. There was an interaction between the effects of carbohydrates and type of protein-free diet on faecal and urinary excretion. The sum of amino acids in faecal protein was the lowest on the PEC diet, but the amino acid composition expressed as a per cent of total amino acid content was similar in all groups. It can be concluded that dietary carbohydrates alter the excretion patterns of endogenous nitrogen in rats in different ways and that this effect is related to the intensity of their fermentation in the hind gut. PMID- 11006828 TI - Effects of vitamin B6 supplementation in rats during lactation on vitamin B6 concentration and transaminase activities in the offspring. AB - The aim of the present investigation was to study the effect of a varying maternal vitamin B6 supplementation during lactation period on vitamin B6 levels in blood, liver and total body, and on the activity of two transaminase enzymes in the offspring. Therefore, eighty female Sprague-Dawley rats were fed a semi synthetic diet (0.2 mg vitamin B6 per kg) which was supplemented during gravidity with 5 mg vitamin B6 per kg diet. During the following lactation period the rats were assigned to one of 10 vitamin B6 treatment groups (supplementation of 0, 3, 6, 9, 12, 15, 18, 36, 360, 3600 mg vitamin B6 per kg diet). At day 14 of lactation the pubs of all dams were decapitated and blood, liver, and carcass were used for analysis of vitamin B6 concentration, activities of two transaminases, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in plasma, erythrocytes, and liver, and of haematological parameters. While the liver and total body wet weights as well as the haematological parameters (red blood cells, haemoglobin concentration, hematocrit, middle corpuscular cell volume, middle corpuscular haemoglobin, middle corpuscular haemoglobin concentration) did not differ within the experimental groups, the present data clearly show that in blood, liver and total body of the offspring exists a slight dose-response relationship between the maternal dietary vitamin B6 supplementation and the vitamin B6 concentration. Concerning the activities of the transaminases a dietary supplementation above 3 mg vitamin B6 per kg diet had no influence on the AST and ALT activities in offspring plasma. In the erythrocytes no statistical significant influence of the vitamin B6 supplementation during lactation on the activities of AST and ALT was found. The activities of ALT and AST in liver were not consistently altered by the vitamin B6 supplementation of the dams during lactation. In conclusion these results indicate that a minimal maternal dietary vitamin B6 supply of 3.1 mg per kg diet is necessary with regard to health and development of their offspring. But not all of the analysed parameters as the liver and total body weights, the activities of AST and ALT in the erythrocytes, and the haematological parameters were influenced by a deficient maternal dietary vitamin B6 supply. PMID- 11006829 TI - Fermentation of carbohydrates and yield of microbial protein in mixed cultures of rabbit caecal microorganisms. AB - Fermentation pattern and yields of microbial protein were investigated in cultures of the rabbit caecal contents supplied with glucose, xylose, starch, pectin and xylan. Rabbits at the age of 4 weeks (before weaning) and 3 months were slaughtered, their caecal contents added at 1.1% to growth media and incubated anaerobically at 39 degrees C for 18 h. Caecal microorganisms of 4-week old rabbits produced no methane and caproate, less butyrate, but more propionate than microorganisms of 3-month-old rabbits. In both groups of rabbits, fermentation of xylose produced significantly more propionate and less butyrate than fermentation of glucose. More propionate and less acetate was formed from starch than from pectin. In caecal cultures from 4-week-old rabbits with pectin, the molar percentages of acetate was significantly higher and percentages of other short-chain fatty acids (SCFA) lower than in cultures with starch or xylan. In cultures from 3-month-old rabbits, fermentation of pectin and xylan produced similar SCFA profiles, different from SCFA molar composition in cultures with starch. Average production of microbial protein was 129 mg per 1 g of carbohydrate digested (range 110 to 141 mg/g). Protein yields were the same on glucose and xylose, but nonsignificantly higher on starch than on pectin and xylan. It can be concluded that the characteristics of substrate affected fermentation pattern in mixed cultures of rabbit caecal microorganisms. Substrate effects on protein yields were not statistically significant, due to high variation. PMID- 11006830 TI - Estimation of net nutrient oxidation and lipogenesis in growing pigs. AB - Data from previous series of trials were reevaluated in order to quantify the net nutrient oxidation and lipogenesis by combining data from balance experiments and indirect calorimetry measurements. The experiments were carried out with eight castrated males of Danish Landrace measured individually from 30 to 100 kg BW. All pigs were fed alternately on high feeding level, near ad libitum and on low feeding level, near maintenance. Oxidation of carbohydrate (OXCHO) and fat (OXF) was calculated from gas exchange measurements. On high feeding level the contribution from OXCHO and OXF to the total heat production was 55 and 30%, respectively, at 30 kg BW and fairly constant from 60 kg BW with OXCHO around 80% and no net OXF. At low feeding level the contribution from OXCHO and OXF was 28 and 63%, respectively, at 30 kg BW, increasing to 52% for OXCHO and decreasing to 37% for OXF at 95 kg BW. The lipogenesis on high feeding level increased from 2.4 to 11.9 MJ/d, while on low feeding level it increased from 2.5 to 3.6 MJ/d at 30 to 95 kg BW. The intake of carbohydrate was too low to cover energy requirement by OXCHO on low feeding level and in the first periods on high feeding level and OXF occurred. In spite of a deficit of carbohydrate for oxidation a part of carbohydrate (2.6 MJ/d, approximately 150 g/d) was used for fat synthesis, indicating an importance of lipogenesis in growing pigs. PMID- 11006831 TI - The influence of potato fibre on exocrine pancreatic secretions and on plasma levels of insulin, secretin and cholecystokinin in growing pigs. AB - The effect of a potato fibre preparation on exocrine pancreatic secretions and on gastrointestinal hormone levels in plasma was studied in three 8 weeks old piglets that were surgically fitted with a jugular vein catheter for blood sampling, a pancreatic duct catheter and a T-shaped duodenal cannula for collection of pancreatic juice. The animals were fed for 2 weeks a control diet (experimental period 1), thereafter for 2 weeks the control diet supplemented with 2% potato fibre (experimental period 2) and for another 2 weeks the control diet again (experimental period 3). Additionally, intraduodenal (i.d.) infusions of the experimental diet, the control diet and potato fibre as well as i.v. infusions of a solution containing cholecystokinin (CCK) and secretin were administered. Potato fibre in the diet evoked in tendency an increase in the volume of secretion of pancreatic juice and a significant increase both in the mean values of the total protein content and total activities of lipase, trypsin and alpha-amylase when compared to the control diet. The i.d. infusion of the control diet, experimental diet and fibre infusate as well as the i.v. administration of the hormone infusate led to a spontaneous secretory response of the exocrine pancreas. Besides gastrointestinal hormones, such as CCK, other factors such as short chain fatty acids may be involved in the regulation of the exocrine pancreas. PMID- 11006832 TI - Development of an in vitro incubation technique for the estimation of the utilizable crude protein (uCP) in feeds for cattle. AB - An in vitro incubation technique based on the first stage of the in vitro digestion technique published by Tilley and Terry (1963) was developed to estimate the utilizable crude protein (uCP) of single feeds and feed mixtures as non ammonia-N after 24 h of incubation. The results of 25 feed samples showed that there was a significant relationship between the uCP values calculated by regression based on in vivo data sets (Y, CP [g.kg-1 DM]) and those measured by the in vitro incubation technique (X, CP [g.kg-1 DM], 24 h incubation): y = 0.85x + 18.0, r2 = 0.84, P < or = 0.001. It was concluded that it can be possible to determine the uCP value of single feeds or feed mixtures by this in vitro incubation technique and to estimate the uCP value of feeds by this regression equation. PMID- 11006833 TI - Investigations on the influence of duodenal histidine infusion on nitrogen and amino acid turnover of growing German Holstein bulls. AB - The effect of a continuous duodenal infusion of L-histidine (His) (8 g/d) on the retention of nitrogen was investigated in two experiments (I, II), each of which was carried out using two young bulls. In Exps. I and II, the animals (150-250 kg BW) were fitted with a re-entrant cannula in the proximal duodenum and were fed diets containing 125 g CP/kg DM and 11.5 MJ ME/kg DM. A third experiment (III) using two young bulls (140-200 kg BW) fitted with a simple T-cannula was carried out infusing 6 g L-His. The animals were fed a low protein diet (94 g CP/kg DM and 11 MJ ME/kg DM). The study was done to find out whether or not L-His is the first limiting amino acid (AA) for growing ruminants. N retention was 28 and 31, 38 and 38, 22 and 24 g/d without L-His infusion and with L-His infusion for Exps. I, II and III, respectively. Both in the experiments with a standard protein supply (I, II) and in the experiment with reduced protein supply (III), no significant differences were found between periods with and without infusion of L His. The utilisation of duodenal NAN varied between 39% and 50% and was also not significantly influenced by the duodenal infusion of L-His. No significant effect was observed on the flow of AA into the duodenum. The faecal excretion of AA was also not significantly influenced by the infusion of L-His. The utilisation of individual amino acids as calculated by the ratio of retained AA to intestinal apparently digested AA, did not differ significantly following the duodenal infusion of L-His. As expected, the utilisation of His decreased. Of the different essential AA, L-His was the most utilised (80%) followed by Arg (72%), Met (60%), Leu (45%) and Lys (44%), during periods without supplementation of L His. It is concluded that the intestinal supply of L-His from the basal diet was sufficient for the potential growth level of animals under these experimental conditions. In all AA present at the proximal duodenum, L-His could have at first a limiting effect on the performance of growing young bulls with high body gain. Arg and Met, but not Lys, could be second or co-limiting AA. PMID- 11006834 TI - Use of health economics for decision-making in complex emergencies. Report of a WHO-FICOSSER meeting. Paris, 16-18 December, 1999. PMID- 11006835 TI - [Comparison of databases of the medical profession: an example in anatomy and pathological cytology]. AB - Medical demographic regulation policy is supported by statistical descriptive and prospective studies on the actual and future supply of medical practitioners. Two main data bases are available in France for these analyses one at the Ministry of Health and the other at the French Medical Council. While both are supposed to be exhaustive and reliable, the official publications of the two institutions provide different data. It is difficult to assess the actual number of French doctors. The task is even more problematic for each individual medical or surgical specialty. Using the medical specialty Pathology as an example, this article compares the quality of the different data bases and underlines some of the structural and dynamic problems which could interfere with the validity of the information. Better cooperation between the different institutions involved would be helpful to contribute to the quality of the data bases and the organization of the medical specialty. PMID- 11006836 TI - [Malaria at the dawn of the millennium: lessons taken from the literature]. AB - The entomological approach has dominated in the past the geographical- epidemiological study of malaria. Nowadays, new approaches are considering aspects such as representations, attitudes to adopt, change in the transmission process according to the ecosystems. The geographical approach here highlighted implies firstly the knowledge of the geo-epidemiology at the small areas love. This approach should also consider the entomological parameters, the environmental factors and the therapeutic behaviors in order to evaluate the efficiency of the action programmes. PMID- 11006837 TI - Phenotypic and genetic characterization of Paecilomyces lilacinus strains with biocontrol activity against root-knot nematodes. AB - Efficient selection of fungi for biological control of nematodes requires a series of screening assays. Assessment of genetic diversity in the candidate species maximizes the variety of the isolates tested and permits the assignment of a particular genotype with high nematophagous potential using a rapid novel assay. Molecular analyses also facilitate separation between isolates, allowing the identification of proprietary strains and trace biocontrol strains in the environment. The resistance of propagules to UV radiation is an important factor in the survival of a biocontrol agent. We have analyzed 15 strains of the nematophagous fungus Paecilomyces lilacinus using these principles. Arbitrarily primed DNA and allozyme assays were applied to place the isolates into genetic clusters, and demonstrated that some genetically related P. lilacinus strains exhibit widespread geographic distributions. When exposed to UV radiation, some weakly nematophagous strains were generally more susceptible than effective isolates. A microtitre tray-based assay used to screen the pathogenic activity of each isolate to Meloidogyne javanica egg masses revealed that the nematophagous ability varied between 37%-100%. However, there was no clear relationship between nematophagous ability and genetic clusters. Molecular characterizations revealed sufficient diversity to allow tracking of strains released into the environment. PMID- 11006838 TI - Stoichiometry of diauxic growth of a xylanase-producing Bacillus strain. AB - In this work, the establishment of material balances and stoichiometry of the growth of Bacillus sp. was undertaken. This strain produces high quantities of a xylanase suitable for use as bleach boost agent in chlorine-free bleaching sequences of paper pulp. As carbon dioxide plays an important role as a growth factor, bacterial growth in two fermentations, one fed with air and another fed with carbon-dioxide-enriched air, were compared. For this purpose, a method permitting the determination of the consumption of the two carbon sources, xylan and peptone, was proposed. The material balances revealed that in both cases, the bacteria first use peptone as their carbon source, and then xylan in the second part of the growth phase. The aerated culture showed diauxic growth on these two substrates, whereas carbon-dioxide-enriched air caused disappearance of the metabolic adaptation phase, and rendered biomass production more economic. The fermentation fed with air needed 30% more xylan than the fermentation fed with carbon-dioxide-enriched air for the same quantity of biomass produced. PMID- 11006839 TI - Effects of experimental conditions on mycorrhizal relationships between Pinus sylvestris and Lactarius deliciosus and unprecedented fruit-body formation of the Saffron milk cap under controlled soilless conditions. AB - The mycorrhizal relationships between pines and two edible species of Lactarius sect. Dapetes were investigated by optimizing the experimental conditions of mycelial growth and of mycorrhizal colonization of pine seedlings. In vitro mycelial growth of Lactarius deliciosus and L. sanguifluus was improved on a buffered medium containing glucose, amino acids, and vitamins. Two methods of mycorrhization of pines with Lactarius deliciosus were tested. The mycorrhizal colonization was rapid and intense under non-aseptic conditions with a low nutrient supply and without exogenous glucose. A positive influence of mycorrhizal colonization on Pinus sylvestris growth was subsequently observed. Under axenic conditions and with a high nutrient supply, mycorrhization was stimulated at 10 g/L of exogenous glucose, irrespective of the phosphorus concentration. At high phosphorus level (1 mM) and 0.1, 1.0, or 10.0 g/L glucose, growth of Pinus sylvestris was reduced by inoculation. Stability and development of Pinus spp./Lactarius deliciosus symbioses were assayed in a climatic chamber using containers filled with a synthetic substrate. Over a 2-year culture period, the root systems of the pine seedlings were heavily colonized by Lactarius deliciosus. One year following inoculation, Lactarius deliciosus fruit-body primordia appeared associated with Pinus sylvestris seedlings. Six months later, two mature basidiomata were obtained. This is the first report of soilless fruit body formation of this edible mushroom. PMID- 11006840 TI - Localization of fungal fimbriae by immunocytochemistry in pathogenic and nonpathogenic isolates of Venturia inaequalis. AB - Pathogenic and nonpathogenic isolates of Venturia inaequalis were grown in liquid culture. Hyphae were treated with two types of fimbrial antiserum (AU- and AV-1) and examined by immunofluorescent microscopy, in order to establish the distribution of fimbrial epitopes in whole cell mounts. The AV-1 antiserum was specific for the glycoprotein subunits while the AU-antiserum was specific for the protein moieties present on the fimbriae of Mycobotryum violaceum. The use of fimbrial antiserum with immunocytochemistry and transmission electron microscopy demonstrated a clear distinction between pathogenic and nonpathogenic isolates of V. inaequalis, based on the appearance of the fungal cell wall and the distribution of fimbrial epitopes labeled with AV-1 antiserum and immunogold complex. In actively growing hyphae of the pathogenic isolate, characterized by distinct cellular organelles, small vacuoles, and lipid bodies, fimbrial epitopes were concentrated in the fungal cell wall and were present minimally on the outer surface. In contrast, actively growing hyphae of the nonpathogenic isolate of V. inaequalis had extensive fine hair-like protrusions in the fungal cell wall which labeled with the AV-1 antiserum and immunogold. The distribution of fimbrial epitopes in V. inaequalis was highly dependent on the developmental growth stage of the fungal mycelium. Aging mycelia in both the pathogenic and nonpathogenic isolates of V. inaequalis were characterized by a large central vacuole and no label. In the pathogenic and nonpathogenic isolates of V. inaequalis grown in vitro, the distribution of fimbrial glycoprotein epitopes provided a more complex profile than that seen in M. violaceum. PMID- 11006842 TI - Physical degradation of wheat straw by the in-vessel and windrow methods of mushroom compost production. AB - Mushroom compost manufacturers in Ireland are moving away from the traditional outdoor phase I windrow method, favouring in-vessel production. Composters and growers have reported better quality compost with faster spawn run and higher yields produced by this process. In the present study, physical examination of samples highlighted differences when comparing the windrow and in-vessel methods of compost production. Observations using scanning electron microscopy suggest that the cuticle of wheat straw from in-vessel production is damaged during phase I, peeling away from the surface in fragments, and exposing the epidermis. Changes in silicon levels on the straw surface acted as a marker for cuticle damage when comparing both composting systems. Cuticle damage may be important during composting and afterwards, as substrate colonisation is faster, and consequently spawn run is shorter. The phase I compost microbial community is altered by the in-vessel technique, producing a predominantly thermophilic bacterial flora in contrast to the mesophilic and thermophilic bacteria and fungi found in windrow phase I compost. These differences may be significant in mushroom compost production. PMID- 11006841 TI - Development of formulations of biological agents for management of root rot of lettuce and cucumber. AB - The effect of various carrier formulations of Bacillus subtilis and Pseudomonas putida were tested on germination, growth, and yield of lettuce and cucumber crops in the presence of Pythium aphanidermatum and Fusarium oxysporum f.sp. cucurbitacearum, respectively. Survival of B. subtilis and P. putida in various carriers under refrigeration (about 0 degree C) and at room temperature (about 22 degrees C) was also studied. In all carrier formulations, B. subtilis strain BACT 0 survived up to 45 days. After 45 days of storage at room temperature (about 22 degrees C), populations B. subtilis strain BACT-0 were significantly higher in vermiculite, kaolin, and bacterial broth carriers compared with other carriers. Populations of P. putida were significantly higher in vermiculite, peat moss, wheat bran, and bacterial broth than in other carriers when stored either under refrigeration (about 0 degree C) or at room temperature (about 22 degrees C) for 15 or 45 days. Germination of lettuce seed was not affected in vermiculite, talc, kaolin, and peat moss carriers, but germination was significantly reduced in alginate and bacterial broth carriers of B. subtilis compared to the non-treated control. Germination of cucumber seed was not affected by any of the carriers. Significantly higher fresh lettuce and root weights were observed in vermiculite and kaolin carriers of B. subtilis compared with P. aphanidermatum-inoculated control plants. Lettuce treated with vermiculite, and kaolin carriers of B. subtilis, or non-inoculated control lettuce plants had significantly lower root rot ratings than talc, peat moss, bacterial broth, and P. aphanidermatum inoculated control plants. Growth and yield of cucumber plants were significantly higher in vermiculite-based carrier of P. putida than the other carriers and Fusarium oxysporum f.sp. cucurbitacearum-inoculated plants. PMID- 11006843 TI - Cholesterol is accumulated by mycobacteria but its degradation is limited to non pathogenic fast-growing mycobacteria. AB - In this report we show that fast-growing non-pathogenic mycobacteria degrade cholesterol from liquid media, and are able to grow on cholesterol as a sole carbon source. In contrast, slow-growing mycobacteria, including pathogenic Mycobacterium tuberculosis and bacillus Calmette-Guerin (BCG), do not degrade and use cholesterol as a carbon source. Nevertheless, pathogenic mycobacteria are able to uptake, modify, and accumulate cholesterol from liquid growth media, and form a zone of clearance around a colony when plated on solid media containing cholesterol. These data suggest that cholesterol may have a role in mycobacterial infection other than its use as carbon source. PMID- 11006844 TI - A physical and genetic map of the Mycoplasma hyopneumoniae strain J genome. AB - A macrorestriction map of the genome of Mycoplasma hyopneumoniae strain J, the type strain of the causative agent of enzootic pneumonia in pigs, was constructed using pulsed-field gel electrophoresis (PFGE) and DNA hybridization. The size of the genome as determined by PFGE was approximately 1070 kb. Assembly of the M. hyopneumoniae genomic map was facilitated and complimented by the simultaneous construction of an ordered cosmid library. Five contigs of overlapping cosmids were assembled, which together represent coverage of approximately 728 kb. Forty two genetic markers (including three types of repeated elements) were placed on the M. hyopneumoniae map. Closer examination of an ApaI restriction fragment contained entirely within a single cosmid insert suggests that the genome size may be overestimated by PFGE. PMID- 11006845 TI - High-frequency interconversion of turbid and clear plaque strains of bacteriophage f1 and associated host cell death. AB - Under normal cultivation conditions, a mixture of turbid and clear plaques is often apparent in cultures of bacterial cells infected with filamentous bacteriophages. Beginning with a culture of wild-type filamentous phage f1, which itself produces turbid plaques, a clear plaque strain (c1) was isolated. From c1, the turbid plaque strain t1 was isolated; from t1, the clear plaque strain c2 was isolated; and from c2, the turbid plaque strain t2 was isolated. Each of these strains was generated with a frequency of approximately 1 x 10(-4). Although filamentous phages have been thought not to induce host cell death, both turbid and clear plaque strains of f1 killed host bacteria. Plating of bacterial cells 1 h after infection revealed that colonies produced by cells infected with either wild-type f1 or strain c2 were smaller than those derived from uninfected cells, and that colony formation by infected cells was reduced by 15% and 38%, respectively. The time course of bacterial growth revealed that, at 4 h after infection, the number of CFU per milliliter of culture of cells infected with wild-type f1 or with strain c2 was reduced by 27% and 95%, respectively, compared with that for uninfected cells. Microculture analysis also revealed that the percentages of nondividing cells in f1 or c2 infected were 19% and 52%, respectively, 4 h after infection with wild-type f1 or with strain c2; no such cells were detected in cultures of uninfected cells. Negative staining and electron microscopy showed that 20% and 61% of cells infected with wild-type f1 or with strain c2 were dead 4 h postinfection. Finally, although the rates of DNA synthesis were similar for infected and uninfected cells, the rates of RNA and protein synthesis were markedly reduced in infected cells. PMID- 11006846 TI - Cloning of the histidine biosynthetic genes from Corynebacterium glutamicum: organization and analysis of the hisG and hisE genes. AB - The physically linked hisG and hisE genes, encoding for ATP phosphoribosyltransferase and phosphoribosyl-ATP-pyrophosphohydrolase were isolated from the Corynebacterium glutamicum gene library by complementation of Escherichia coli histidine auxotrophs. They are two of the nine genes that participate in the histidine biosynthetic pathway. Molecular genetics and sequencing analysis of the cloned 9-kb insert DNA showed that it carries the hisG and hisE genes. In combining this result with our previous report, we propose that all histidine biosynthetic genes are separated on the genome by three unlinked loci. The coding regions of the hisG and hisE genes are 279 and 87 amino acids in length with a predicted size of about 30 and 10 kDa, respectively. Computer analysis revealed that the amino acid sequences of the hisG and hisE gene products were similar to those of other bacteria. PMID- 11006847 TI - Purification and characterization of a 4-hydroxybenzoate decarboxylase from an anaerobic coculture. AB - The oxygen-sensitive 4-hydroxybenzoate decarboxylase (4OHB-DC) activity from a phenol-carboxylating coculture, consisting of Clostridium-like strain 6 and an unidentified strain 7, was studied. Assays done with cell extracts showed that the optimal pH was 5.0-6.5 and the Km was 5.4 mM. The activity decreased by 50% in the presence of 5 mM EDTA, and it was restored and even enhanced by the addition of Mg++, Mn++, Zn++, or Ca++. After purification, the molecular mass of the enzyme was estimated as 420 kDa by gel chromatography, and as 119 kDa by SDS PAGE, suggesting a homotetrameric structure. Its pI was 5.6. The N-terminal amino acid sequence showed 95% and 76% homology with the pyruvate-flavodoxin oxidoreductase (nifJ gene product) from Enterobacter agglomerans and Klebsiella pneumoniae, respectively. The purified enzyme also slowly catalyzed the reverse reaction, that is the phenol carboxylation. These characteristics suggest that this enzyme is different from other known decarboxylases. This includes the 4OHB DC from Clostridium hydroxybenzoicum, which is the only one that had been purified before. PMID- 11006848 TI - Production of cellulases and xylanases by low-temperature basidiomycetes. AB - Three of four isolates, representing phylogenetically distinct groupings of low temperature basidiomycetes (LTB), were capable of utilizing wheat straw, and to a lesser extent conifer wood at 15 degrees C. A cottony snow mould LTB (LRS 013) and a fruit rot LTB (LRS 241) grown on straw significantly degraded filter paper, carboxymethylcellulose (CMC), p-nitrophenyl beta-glucopyranoside (i.e., beta glucosidases), and xylan. Enzymes produced by Coprinus psychromorbidus (LRS 067) were limited to xylanases from straw and wood and beta-glucosidases from wood. A sclerotia-forming LTB (LRS 131) exhibited poor growth on both substrates, and did not produce detectable quantities of extracellular enzymes. None of the LTB isolates tested degraded avicel. The temperature optima of CMCases and xylanases in the filtrates from the straw medium ranged from 25 degrees C to 55 degrees C, and with the exception of LRS 067, significant activity was observed at 5 degrees C. Two cellulases (25 and 31 kDa) and two xylanases (24 and 34 kDa) were observed on zymograms for LRS 013 and 241. Reduction of enzymes with 2-mercaptoethanol adversely affected their activity on zymograms, and an additional cellulase band was observed for non-reduced samples. This study indicates that LTB produce an array of cellulolytic and xylanolytic enzymes, and that some of these enzymes possess low-temperature optima which may facilitate degradation of plant fibre under low-temperature conditions. PMID- 11006849 TI - Photo quiz: urticaria pigmentosa. PMID- 11006850 TI - What's eating you? Pulex irritans. PMID- 11006851 TI - Multiple fire ant stings: report of 3 cases and review of the literature. AB - Imported fire ant sting reactions are becoming an increasing problem in the United States. It is important for clinicians to be familiar with their possible cutaneous and noncutaneous presentations. We present 3 cases with multiple fire ant stings, followed by a review of the literature. PMID- 11006852 TI - Subacute cutaneous lupus erythematosus: report of a patient who subsequently developed a meningioma and whose skin lesions were treated with isotretinoin. AB - Cancer has been reported in patients with systemic lupus erythematosus (SLE). A possible association of the development of hematologic malignancies in patients with SLE has been suggested. In some patients, subacute cutaneous lupus erythematosus, a distinct subset of lupus erythematosus, has appeared, resolved, or both as a solid tumor-related paraneoplastic syndrome. A woman in whom a meningioma was diagnosed 44 years following the onset of subacute cutaneous lupus erythematosus is described; her skin lesions improved after starting isotretinoin therapy. The relationship between lupus erythematosus and neoplasia is summarized and the management of subacute cutaneous lupus erythematosus with retinoids is reviewed. PMID- 11006853 TI - Spiny keratoderma. AB - Spiny keratoderma is a descriptive term used to encompass a variety of unusual, disparate keratodermas. Spiny keratoderma has been associated with lipid abnormalities and has been limited to the palms and soles in some individuals. We describe an acquired case of spiny keratoderma in which an adult woman developed filiform lesions predominating on the trunk and proximal extremities. Treatment with topical emollients and keratolytic agents was unsuccessful, but topical tazarotene led to long periods of resolution. She has had no other associated abnormalities. The clinical features and differential diagnosis of spiny keratoderma are reviewed. PMID- 11006854 TI - Rapidly progressive fatal cutaneous T cell lymphoma with a trauma-related presentation. AB - A case of rapidly progressive cutaneous T cell lymphoma with a trauma-related presentation in a 73-year-old man is reported. Clinically, the patient presented with an ulcerated cutaneous mass at the site of trauma-related hematoma of the leg. The histopathology was that of tumor phase cutaneous T cell lymphoma with involvement of the skin and subcutis. The diagnostic challenge of this clinical presentation and the rapidly progressive course are highlighted. PMID- 11006855 TI - Kimura's disease presenting as subcutaneous facial plaque in an African American. AB - Kimura's disease is a benign, uncommon, chronic inflammatory condition that usually presents with painless subcutaneous nodules or plaques in the head and neck region. Although the disease is predominantly found in Asian populations, there are occasional cases reported among Caucasians and rare occurrences in African populations. The etiology and pathogenesis of the disease are unknown and the clinical presentation can mimic several benign and malignant disease states. The accurate diagnosis of Kimura's disease is based on clinical and histopathological findings. There is no evidence of malignant transformation and occasional spontaneous resolution occurs. Various treatment modalities have been suggested in the management of this condition. Oral corticosteroids have been the mainstay of therapy, even though steroid withdrawal can result in lesion recurrence. PMID- 11006856 TI - Disseminated cryptococcosis presenting as pseudofolliculitis in an AIDS patient. AB - We report the case of a 42-year-old man with AIDS and an unusual presentation of disseminated cutaneous cryptococcosis. The eruption was characterized by excoriated papules of the upper body and was initially diagnosed as folliculitis. A pseudofollicular eruption is a rare presentation for disseminated cryptococcosis. PMID- 11006857 TI - Multiple onychocryptosis following treatment of onychomycosis with oral terbinafine. AB - The authors report an unusual case of multiple onychocryptosis, which developed following treatment of onychomycosis with oral terbinafine. With new growth of the healthy nail plate, the distal aspect of multiple toenails became ingrown with periungual inflammation. This required several minor surgical procedures to alleviate the onychocryptosis. The authors present this case report as a potential complication of oral antifungal therapy. PMID- 11006858 TI - Superficial granulomatous pyoderma. AB - We report the case of a 30-year-old man who, from the age of 16, presented with abscess-type lesions that turned into ulcers with a torpid course and with histopathologic characteristics of superficial granulomatous pyoderma. The patient has been followed for 6 years, during which time the only treatment necessary to prevent recurrence of lesions has been the administration of different doses of minocycline. PMID- 11006859 TI - Herpes zoster in the medically healthy child and covert severe child abuse. AB - Herpes zoster is associated with depressed cell-mediated immunity and occurs rarely in the medically healthy nonimmunocompromised child. We report 4 cases of childhood-onset herpes zoster in the absence of a medical disorder. All 4 patients reported experiencing severe, chronic child abuse when the herpes zoster first appeared. It is possible that the severe chronic psychologic stress resulting from the abuse depressed the patients' cell-mediated immune status and thereby predisposed them to herpes zoster. Our findings suggest that the clinician's suspicion should be heightened for the possibility of covert child abuse and secondary stress when managing an otherwise apparently healthy child with herpes zoster. PMID- 11006860 TI - [Johann Nepomuk Czermak as a neurophysiologist in Leipzig]. AB - Johann Nepomuk Czermak (1828-1873) gained a lasting reputation in the fields of laryngology, rhinology and dentistry. However, his works in the discipline he liked most--neurophysiology--have almost been forgotten. This paper focuses on those articles, which Czermak wrote during his last years in Leipzig (from 1869). Also it was here that he was finally able to fulfill his dream of a "spectatorium", a perfect place for both research and teaching at that time. This institution was the perfect expression of Czermak's uncompromisingly scientific approach, his views in social as well as matters of teaching. It is part of his tragedy though, that Czermak died too early in Leipzig, the place where he could finally realize many of his scientific visions, and that his inheritance was soon forgotten. PMID- 11006861 TI - [Positron emission tomography in diagnosis and prognosis of postanoxic cerebral dysfunctions]. AB - Clinical signs, laboratory tests, EEG or evoked potentials only permit an indirect estimation of the extent of structural brain damage following severe global brain anoxia. Positron emission tomography (PET) permits additional insights into the extent of neuronal damage in acute and persistent postanoxic vegetative states (VS). PET documents a severe and irreversible damage of supratentorial cortical structures in postanoxic VS and allows its differential diagnosis from related disorders, e.g. the locked-in-syndrome. PET clearly distinguishes functional alterations in VS from those in non-REM sleep and documents that patients in VS are not in a sleep-like condition, but in a state closely related to deep anesthesia. Furthermore, the extent of impairment of the residual cortical glucose consumption yields information concerning the possible recovery of consciousness and neuronal function in VS. In combination with clinical, laboratory and neurophysiological findings, PET may be helpful to establish the individual prognosis in acute VS. PMID- 11006862 TI - [The EEG as an indicator of cholinergic deficit in Alzheimer's disease]. AB - There are tight relationships between the EEG and the activity of the cholinergic system. The excitability of cortical neurons is increased by the ascending cholinergic projections of the nucleus basalis of Meynert. Cholinergic deafferentiation of the cortex leads to an increase of slow-wave EEG activity. Cholinergic deficit is a typical feature of Alzheimer's disease. It is goes along with characteristic EEG alterations, mainly increased activity in the slow frequency bands. The amount of EEG alterations increases as the disease progresses. It is correlated with the severity of symptoms, neuropsychological performance, cerebral perfusion, cerebral glucose utilization and the extent of histopathological changes. EEG alterations are a valuable diagnostic tool in Alzheimer's disease. They may be suited for early diagnosis and may allow the prediction of the therapeutic response to cholinergic drugs. PMID- 11006863 TI - [The effect of the menstruation cycle on manifestations of pychiatric diseases]. AB - In the present study we investigated whether a correlation exists between menstrual cycle phase on the day of an acute psychiatric admission and diagnostic entities. Therefore we assessed the menstrual cycle phase in 155 women at the time of acute admission for any non-organic psychiatric disorder. A specific diagnosis according to ICD-10-criteria and to Leonhard's nosology was established without knowledge of the menstrual cycle phase. Independent of diagnosis and classification, the majority of patients (57%) was admitted during the pre menstrual/menstrual period. Comparing the frequencies of admission before (increasing blood-estrogen-level) and after ovulation (decreasing blood-estrogen level) we found using ICD-10 criteria there were no significant differences between affective psychoses (F3), acute polymorphous psychotic disorder (F23), schizophrenia/schizoaffective psychoses (F20 and F25) and patients suffering from neuroses or personality disorders (F4-F6). Applying Leonhard's criteria we found no significant differences between endogeneous psychoses and personality disorders and no significant differences between cycloid psychoses and affective psychoses or affective psychoses and schizophrenias as well. However, patients with cycloid psychoses were significantly more frequently admitted to hospital during the luteal-/menstrual phase than patients with schizophrenia (chi 2-Test, p = 0.02). These findings do not confirm a specificity of a pre-menstrual exacerbation of psychotic symptoms for schizophrenia. Rather we found cycloid psychoses to be significantly more frequently associated with premenstrual exacerbation of symptoms. PMID- 11006864 TI - [Normative needs for care of schizophrenics: a useful concept for community psychiatric planning?]. AB - One step in a public health research project focuses on the analysis of the individual (expert-based) normative needs for mental health care of chronic schizophrenic patients (n = 115) in the Dresden Region during the first year post hospital release and the extent to which this can be met by the current established level of complementary care. It is an exemplary contribution to the evaluation of community psychiatry as restructured in the Free State of Saxony following German reunification. The results of the study can be condensed to the following interpreting essential statements. Schizophrenics' normative needs for care are not a statistical issue. The single case analysis corroborates a high rate of relevant fluctuations, above all within the clinical sector (e.g. concerning "dyskinesias and other side effects"), that pose a particular challenge to the flexibility of a system of community psychiatry. This includes that the consequence for the practice of care implies then that when diagnosing course, attention must be paid to shifts in the content of the needs for mental health care (e.g. increasing importance of factors, which contain impairments of basic social competences) in order to orient to them any health care measures already initiated.--With the aid of the used research instrument (Needs for Care Assessment) deficits in meeting the needs for care can be identified. In the Dresden Region considerable deficits persist apparently in the subsections recreational activities and occupational and communication skills, which can be ascribed to the lack of appropriate institutions of care in the area.--The normative needs for care of schizophrenic patients cannot be determined simply on the basis of a few, quickly identifiable markers. Rather it demands individualized analysis incorporating variables pertaining to psychopathology, subjective coping, social competence and the course of the disorder. The development of the needs for care over the period of one year can apparently be predicted by trends in the social sector that are already visible within the first months. With regard to aspects of care planning this finding illustrates the limited ability of cross-section surveys to make definitive statements, as well as the predominance of social disabilities over the entire spectrum of the normative needs for psychiatric care. PMID- 11006866 TI - [Communications of the Viktor von Weizscker Society e.V]. PMID- 11006865 TI - [Political change in East Germany 1989-90 in the lives of chronic schizophrenic patients]. AB - Theoretical models suggest a higher vulnerability of chronic schizophrenic patients for critical life events and rapid change of objective living circumstances. On the basis of these models one may hypothesise that the political change in East Germany in 1989/90 was objectively or subjectively distressing to such patients and had a negative impact on their illness. In a retrospective longitudinal study, we investigated patients' assessment of political change, life events that were potentially related to the political changes, impact of the changes on subjective quality of life, and hospitalization rates during five year periods prior to and following the change. 120 patients with chronic schizophrenia in East Berlin and 70 in Chemnitz were examined using quantitative and qualitative methods. Patients reported more individual freedom and better care after 1989. They complained about stress due to loss of employment, financial disadvantages and rising crime rates. Despite the occurrence of potentially change-related life events after 1989, patients stated very little impact of the changes on their illness. Retrospectively, patients in East Berlin reported a significantly better subjective quality of life in 1994 than in 1984. Hospitalization rates did not increase after 1989. The findings are not consistent with the hypothesis and do not suggest that political changes in East Germany in 1989/90 had--subjectively or objectively--a substantial negative effect on the illness of patients with chronic schizophrenia. PMID- 11006867 TI - [Gastrointestinal oncology]. PMID- 11006868 TI - [Barrett esophagus: diagnosis and treatment]. PMID- 11006869 TI - [Gastric carcinoma]. PMID- 11006870 TI - [Gastrointestinal lymphoma: etiology, pathogenesis and therapy]. PMID- 11006871 TI - [Gallbladder and bile duct carcinoma]. PMID- 11006872 TI - [Pancreatic carcinoma]. PMID- 11006873 TI - [Colon carcinoma: early detection and endoscopic prevention]. PMID- 11006874 TI - [Primary prevention of sporadic colorectal carcinoma by diet modification and drugs?]. PMID- 11006876 TI - [Emergencies due to drug abuse]. PMID- 11006875 TI - [Colon carcinoma. Consensus of therapeutic strategies]. PMID- 11006877 TI - [Multiple muscle abscesses in a Tibetan woman]. PMID- 11006878 TI - [Initial diagnosis of Caroli disease in a 71-year old man]. PMID- 11006879 TI - [Seeding metastasis caused by fine needle biopsy?]. PMID- 11006880 TI - [High HbA1c level in presence of good blood glucose control]. PMID- 11006881 TI - [Angiotensin-II-receptor antagonists for cardiac insufficiency]. PMID- 11006882 TI - [Peripheral neuropathy in diabetes mellitus from the perspective of internist]. PMID- 11006883 TI - [28-year old patient requiring reanimation for heart arrest after hospitalization for deep vein thrombophlebitis]. PMID- 11006884 TI - Wall stress and the heart. PMID- 11006885 TI - Experimental studies on myocardial stretch and ventricular arrhythmia in hypertrophied and non-hypertrophied hearts. AB - Hypertension affects about 5% of western populations and in the majority of cases it is of unknown aetiology. It exposes the heart to greater levels of myocardial stretch as a result of increased systolic pressure and peripheral resistance. Under certain circumstances myocardial stretch may trigger arrhythmias but the mechanisms and clinical importance of this phenomenon are unclear. This article outlines the risks of sudden cardiac death conferred by hypertension and left ventricular hypertrophy, presents the results of experiments using an animal model of myocardial stretch and discusses some possible mechanisms underlying stretch-induced arrhythmias which may be important in hypertensive patients. PMID- 11006886 TI - Mechanoelectric coupling as a fundamental surrogate in ventricular arrhythmia? PMID- 11006887 TI - Wall stress and hypertension. AB - The precise role that abnormal wall stress may play in the pathophysiology of hypertensive heart disease is not known. Hypertension is almost unique in that it ultimately affects all parts of the law of Laplace equation, i.e. intraventricular pressure changes and with the advent of left ventricular hypertrophy both internal radius and wall thickness alter. If heart failure supervenes the components of the equation change once more. This article will discuss the implications of abnormal wall stress at these various stages in hypertensive heart disease. PMID- 11006888 TI - Combined seropositivity for H. pylori and C. pneumoniae is associated with age, obesity and social factors. AB - BACKGROUND: Manifestations of cardiovascular disease (CVD) have been associated with chronic infection by Helicobacter pylori and Chlamydia pneumoniae both in cross-sectional and in prospective follow-up cohort studies. This association may be partly due to an increase in metabolic risk factors for CVD, secondary to low grade inflammation caused by infections. OBJECTIVE: To investigate for subjects classified according to serology titres for infection with C. pneumoniae and H. pylori associations between seropositivity and the degree of obesity and fasting insulin levels, as well as social factors. METHODS: Using methods based on those in earlier investigations of hypertensive patients in the Dalby primary-health care district, southern Sweden, we investigated frozen samples from serum of 310 middle-aged treated hypertensives and 288 age-matched and sex-matched normotensive controls from a defined population. The baseline examination included the measurement of weight, height and blood pressure as a mean of two office readings with the subject supine. The body mass index (BMI) was calculated as kg/m2. Fasting blood samples were drawn for measurements of levels of serum lipids, blood glucose, plasma insulin and serum lipids, including total cholesterol and triglycerides. The serology titres for H. pylori were determined by an enzyme-linked immunosorbent assay. The titres for C. pneumoniae were determined by a micro-immunofluorescence method. Self-reported factors concerning social and lifestyle backgrounds were recorded. RESULTS: The group (n = 245) of subjects with combined positive serology for H. pylori and C. pneumoniae differed from the group without any positive serology (n = 57) in age (61.6 versus 57.4 years, P < 0.05) and BMI (27.3 versus 25.8 kg/m2, P < 0.05). The seropositive group also differed in terms of fasting levels of insulin (12.7 versus 11.6 pmol/l, P < 0.05), but this difference did not remain significant after adjustment for age and BMI. We detected no intergroup difference in blood pressure and levels of glucose and lipids. Members of the group with combined seropositivity reported having a lower social-class position (educational level) than that of members of the seronegative group. CONCLUSION: Subjects with combined positive serology for H. pylori and C. pneumoniae are characterized by greater age, lower social class and higher BMI, as well as higher fasting levels of insulin than those of seronegative subjects. Obesity might be a marker not only for lower social class but also for greater than normal susceptibility to such infections. PMID- 11006889 TI - A common polymorphism in methionine synthase reductase increases risk of premature coronary artery disease. AB - BACKGROUND: Methionine synthase reductase (MTRR) catalyzes the regeneration of methylcobalamin, a cofactor of methionine synthase, an enzyme essential for maintaining adequate intracellular pools of methionine and tetrahydrofolate, as well as for maintaining homocysteine concentrations at nontoxic levels. We recently identified a common A-->G polymorphism at position 66 of the cDNA sequence of MTRR; this variant was associated with a greater than normal risk for spina bifida in the presence of low levels of cobalamin. OBJECTIVE: To investigate whether the polymorphism was associated with alterations in levels of homocysteine, folate, and vitamin B12, and with risk of developing premature coronary artery disease (CAD), in a population of individuals presenting for cardiac catheterization procedures. METHODS: We screened 180 individuals aged < 58 years with angiographically documented coronary-artery occlusions or occlusion free major arteries for the presence of the 66A-->G MTRR polymorphism using a polymerase-chain-reaction-based assay. RESULTS: We identified a trend in risk of premature CAD across the genotype groups (P = 0.03) with a sex-adjusted relative risk of premature CAD equal to 1.49 (95% confidence interval 1.10-2.03) for the GG versus AA genotype groups. There was no difference in fasting levels of plasma total homocysteine, serum folate, and vitamin B12 among the three MTRR genotypes. CONCLUSIONS: Our findings suggest that the GG genotype of MTRR is a significant risk factor for the development of premature CAD, by a mechanism independent of the detrimental vascular effects of hyperhomocysteinemia. This association needs to be confirmed in other studies. PMID- 11006890 TI - Multifactorial approach to the prevention of coronary heart disease: from computer to paper and pencil? AB - BACKGROUND: In Europe the multifactorial clinical approach to the prevention of coronary heart disease is based on the Framingham equation presented in graphical form including age, sex, level of total serum cholesterol, systolic blood pressure and smoking. OBJECTIVE: To propose a straightforward paper-and-pencil score (Global Coronary Risk Score) including level of high-density lipoprotein cholesterol for the Belgian or more broadly western European population derived from 10-year follow-up mortality of a Belgian national population sample. RESULTS: This score has the same predictive power as the Framingham equation both for men aged 35-74 years and for women aged 50-74 years. It gives a ranking of subjects into four groups according to their relative risks. CONCLUSION: Coronary Risk Score is user friendly and probably has pedagogical virtues. PMID- 11006891 TI - High prevalence of seropositivity for antibodies to Chlamydia-specific lipopolysaccharide in patients with acute coronary syndrome. AB - BACKGROUND: Results of recent studies have demonstrated that there is an association between infection with Chlamydia pneumoniae and coronary artery disease (CAD). Inflammatory response caused by chlamydial infection has been considered to contribute to the development of atherosclerosis in coronary arteries. OBJECTIVE: The aim of this study was to investigate the specific relations between chlamydial infection and coronary events in patients with CAD. METHODS: We measured serum levels of immunoglobulin A and G antibodies against Chlamydia spp.-specific lipopolysaccharide in 155 patients with CAD and 60 age matched and sex-matched healthy controls by enzyme-linked immunosorbent assay. CAD patients were divided into groups of the patients with acute coronary syndrome [(ACS), n = 35], old myocardial infarction [(OMI), n = 60] and chronic coronary heart disease [(CCHD), n = 60]. RESULTS: Prevalence of both seropositive antibodies in the control group and CCHD group were not different. In contrast, in ACS group there were significantly higher prevalences of seropositive immunoglobulin A (46 versus 12%, P = 0.0001) and G (74 versus 45%, P = 0.005) antibodies and in OMI group there was a significantly higher prevalence of seropositive immunoglobulin A antibodies (28 versus 12%, P = 0.02). Furthermore, compared with CCHD group, in ACS group there were significantly higher prevalences of seropositive immunoglobulin A (P = 0.00006) and G (P = 0.002) antibodies and in OMI group there was a higher prevalence of seropositive immunoglobulin A (P = 0.01). Adjustment for confounding factors did not change these findings. CONCLUSIONS: Infection with Chlamydia is significantly associated with ACS and OMI, but not with CCHD. These findings suggest that chronic and reactive infection with Chlamydia can lead to disruption of vulnerable plaque in patients with ACS. PMID- 11006892 TI - Risk factors for atherosclerosis in young individuals. AB - Atherosclerosis starts in childhood, and is accelerated in some individuals. A cluster of clinical and biochemical factors constitute the risk profile for many of them, perhaps most important being metabolic insulin resistance syndrome. Insulin resistance and its components for children and adolescents, especially obesity and dyslipidemia, are generators of hypertension, glucose intolerance and complications of atherosclerosis in adulthood. Some individuals are genetically predisposed, particularly those with the family history of such disorders. For many subjects, there is 'tracking' of metabolic and lifestyle factors from early age to adulthood. Several new risk factors of atherosclerosis (e.g. level of lipoprotein (a), procoagulant state, hyperhomocysteinemia, low birth weight and adverse in-utero environment, and possibly inflammatory markers) are current and potentially future areas of research concerning children and young individuals. Definition of and research on new and hitherto not investigated factors and formulation of strategies to neutralize the known factors are of paramount importance for primary prevention of atherosclerosis. Simple and effective measures for prevention include increasing awareness of the diseases, maintenance of ideal body weight, regular physical exercise, avoidance of smoking and chewing of tobacco, eating a balanced diet, and early periodic monitoring of blood pressure and metabolic status. These measures, starting from childhood, should be applied to all and in particular to the susceptible offspring, predisposed individuals, and populations. PMID- 11006893 TI - Bibliography. Current world literature. PMID- 11006894 TI - Hyperlipidaemia and primary prevention of coronary heart disease: are the right patients being treated? AB - BACKGROUND: In 1997, the Standing Medical Advisory Committee report suggested that patients with a coronary heart disease risk of 3% per year or greater should be considered appropriate for lipid-lowering medication. The report stated that cholesterol concentration alone is a poor predictor of absolute risk of coronary heart disease and recommended the Sheffield table as a method of estimating the coronary heart disease risk. OBJECTIVE: To assess the impact of the Standing Medical Advisory Committee report on the management of patients with hyperlipidaemia in the primary prevention of coronary heart disease in primary care. METHOD: A survey questionnaire giving the clinical details of 20 patients with various coronary heart disease risk factors was sent to 200 general practitioners in the West Midlands, UK. RESULTS: Forty-eight percent of the respondents used clinical assessment/perception as the sole means of risk assessment and 26% used the Sheffield table. In patients who did not require treatment, 40.1% of the decisions were inappropriate and, in patients who required treatment, 35.1% of the decisions were inappropriate. Overall, inappropriate decisions were made in 37.9% of the responses. Despite the clear advice in the Standing Medical Advisory Committee report on the importance of incorporating multiple risk factors in estimating absolute coronary heart disease risk, only total cholesterol and triglycerides were significant in influencing treatment decisions. CONCLUSIONS: The Standing Medical Advisory Committee recommendations on the management of hyperlipidaemia in primary prevention of coronary heart disease are not widely used. Large savings could be made by correctly identifying and treating individuals at high risk. We recommend use of the full Framingham risk score in assessment of coronary heart disease risk in primary care. PMID- 11006895 TI - Re-survey of the Whitehall study of London civil servants: changes in risk factors for cardiovascular disease during 29 years of follow-up. AB - BACKGROUND: Substantial uncertainty persists about the relevance of blood pressure and cholesterol to the risk of cardiovascular disease in the elderly. OBJECTIVE: To investigate the determinants of cardiovascular risk in old age, and the relevance of such risk factors when recorded in middle and old age. METHODS: A re-survey in 1997 of 8537 survivors of a cohort of men who were originally examined in 1967-1970 when aged 40-69 years. RESULTS: Completed questionnaires were received from 7050 (82%) of the survivors, and blood pressure and blood samples from 5427 (64%). The response rate declined with increasing age, was inversely related to markers of socioeconomic status in 1967-70 and in 1997, and was lower in those who had been current smokers or had a higher blood pressure level in 1967-70. After excluding those with reported cardiovascular disease (25% of respondents), the mean levels of total cholesterol and apolipoprotein B were lower in older age groups, whereas apolipoprotein A1 levels did not vary much with age. Among those with risk factors recorded both in 1967-70 and 1997, the prevalence of smoking had declined by two-thirds (32% in 1970 and 12% in 1997), the prevalence of diabetes had increased (0.3% versus 4.5%), and the mean systolic blood pressure had increased by 16 mmHg (130 versus 146 mmHg), but the diastolic blood pressure had not changed materially (80 versus 81 mmHg), and the measured levels of total cholesterol had increased by 0.5 mmol/l (although that change may be artefactual). CONCLUSION: Follow-up of vital status in this cohort should permit an assessment of the relevance of risk factors recorded in middle and old age to cardiovascular disease in old age. PMID- 11006896 TI - Leisure-time physical activity and coronary risk factors in women. AB - BACKGROUND: Evidence that physical activity is cardioprotective in women is not as strong as that observed in men. Furthermore, the extent to which exercise protects against coronary heart disease via its influence on classical risk factors remains unclear. This study examines the relationship between reported physical activity, a range of coronary heart disease risk factors and a 10-year predicted coronary heart disease risk score. METHODS: A cross-sectional study of 14,077 female employees aged 30-64 years who were screened between 1988 and 1991 was employed. Measurements included systolic and diastolic blood pressure, body mass index, serum total cholesterol, high-density lipoprotein cholesterol, total cholesterol:high-density lipoprotein cholesterol ratio, low-density lipoprotein cholesterol, triglycerides, apolipoprotein A1, apolipoprotein B, lipoprotein a and fasting blood glucose. Participants were divided into three groups according to reported average frequency of vigorous leisure-time physical activity, that is zero, one to two, or three or more episodes per week. RESULTS: Increasing activity frequency was associated with lower systolic and diastolic blood pressure, total cholesterol, total cholesterol:high-density lipoprotein cholesterol ratio, triglycerides and body mass index (all P < 0.001), low-density lipoprotein cholesterol (P = 0.003), apolipoprotein B (P = 0.04) and blood glucose (P = 0.01) and higher high-density lipoprotein cholesterol (P < 0.001) and apolipoprotein A1 (P = 0.03). There was no association with lipoprotein a. After controlling for possible confounders, these relationships remained statistically significant except for apolipoprotein B and glucose. The odds ratios for being in the top quintile of predicted 10-year coronary heart disease risk for individuals in each category of activity were 1.0 (inactive), 0.70 (one to two episodes of activity per week) and 0.77 (three or more episodes of activity per week). CONCLUSION: Women engaging in vigorous, leisure-time physical activity have a less atherogenic coronary heart disease risk factor profile than those who do not, which translates into a potential reduction of approximately 30% in coronary heart disease risk. PMID- 11006897 TI - Respiratory decline in smokers and ex-smokers--an independent risk factor for cardiovascular disease and death. AB - BACKGROUND AND OBJECTIVE: Although smoking is associated with an increased incidence of cardiovascular disease and death, many smokers remain healthy after many years of smoking. Our objective was to assess whether this variation is related to rate of decline of respiratory function. DESIGN: This was a population based cohort study, its subjects being men born in 1914 from Malmo, Sweden. METHODS: All 291 smokers who since the baseline examination in 1969 had remained in Malmo were invited to a follow-up examination in 1982. Of the 242 participants, 199 men without history of myocardial infarction or stroke were included in the study. Eighty-four of them had quit smoking. The incidence of cardiovascular disease and death during 14 years was studied in relation to the decline in lung function [forced expiratory volume during 1 second (FEV1.0) and vital capacity] between 55 and 68 years of age. RESULTS: Fifty-nine (51%) smokers and 43 (51%) ex-smokers died. Forty-four (38%) smokers and 29 (35%) ex-smokers suffered a cardiovascular event. The mortality rate among smokers in the high, middle and low thirds with regard to the decline in FEV1.0 was 66.5, 44.0, and 37.6, respectively, per 1000 person-years (P for trend = 0.04). The corresponding figures in ex-smokers were 88.7, 42.0, and 35.1 (P for trend = 0.002). The cardiovascular event rate among smokers in these three groups was 56.0, 41.0, and 22.7 events, respectively, per 1000 person-years (P for trend = 0.01). The association remained significant after adjustments for potential confounders. A change in vital capacity was associated with a similar pattern of disease and death. CONCLUSION: Although smoking is associated with an accelerated respiratory decline, there are marked differences between smokers. The increased cardiovascular event and death rates among those whose lung function declined the most suggests that the change in respiratory function can be used as a measure of individual susceptibility. PMID- 11006898 TI - Work stress and coronary heart disease. AB - BACKGROUND: Coronary heart disease (CHD) has been often regarded as a psychosomatic illness; one of the psychosocial variables influencing the onset or course of disease is stress. Work stress in particular may therefore be a significant factor influencing CHD. METHOD: This descriptive review derives from those predictive studies of work stress and CHD published from 1990-2000 based on structured literature searches. RESULTS: There is reasonably robust and consistent empirical evidence indicating some causal relationship between work stress and CHD risk. CONCLUSIONS: Work stress has clear implications for the health and welfare of employees and medico legal implications for employers. PMID- 11006899 TI - Socioeconomic status, ABO phenotypes and risk of ischaemic heart disease: an 8 year follow-up in the Copenhagen Male Study. AB - OBJECTIVES: The association of socioeconomic status with the risk of ischaemic heart disease is only partly explained by the uneven distribution of conventional risk factors. We tested the hypothesis that an uneven socioeconomic distribution of ABO phenotypes could contribute to the explanation. DESIGN: A prospective study controlling for age and other relevant potential confounders: smoking, physical activity, wine consumption, height, weight, serum lipids, blood pressure, hypertension, type II diabetes, serum selenium concentration and soldering fumes exposure. SETTING: The Copenhagen Male Study, Denmark. STUDY PARTICIPANTS: Two thousand, nine hundred and ninety-three men aged 53-74 years without overt ischaemic heart disease. MAIN OUTCOME MEASURE: Incidence of ischaemic heart disease in an 8-year follow-up. RESULTS: Two hundred and forty two men (8.1%) had a first ischaemic heart disease event. There was no association between socioeconomic status and the ABO blood group phenotypes and, in accordance with this, ABO phenotype was not a confounder for the association of socioeconomic status with the risk of ischaemic heart disease. However, ABO blood group was a strong risk or effect modifier. Only among men with the O phenotype was socioeconomic status (social classes IV and V versus social classes I, II and III) associated with a significant excess risk (relative risk 1.7, 95% confidence interval 1.1-2.7 and P = 0.02 after adjustment for confounders; the corresponding relative risks among the A and B/AB phenotypes comparing low social classes with the higher social classes were 1.08 (P = 0.77) and 1.08 (P = 0.89), respectively). CONCLUSION: ABO phenotypes did not contribute directly to the explanation of socioeconomic inequalities in the risk of ischaemic heart disease. However, the finding of ABO phenotypes being effect modifiers for the association of socioeconomic status with the risk of ischaemic heart disease may open up new possibilities of clarifying the roles of socioeconomic status and ABO blood group as cardiovascular disease risk factors. PMID- 11006901 TI - Bibliography. Current world literature. PMID- 11006900 TI - Plasma-induced endothelial activation associated with incident atherosclerosis: prospective results from the Bruneck Study. AB - BACKGROUND: Whether systemic inflammation is an epiphenomenon of atherosclerosis or whether it is part of the atherosclerosis causal pathway requires further study. DESIGN: As part of a prospective population survey on the course and aetiology of atherosclerosis, we investigated the effects of plasma on the endothelial monolayers inducing activation for leukocyte transmigration. METHODS: An age- and sex-stratified random sample of inhabitants of Bruneck (Italy) with and without atherosclerotic disease aged 50-69 years was selected. Carotid arteries were evaluated by duplex sonography at baseline (1990). Carotid arteries were re-evaluated for the development of new plaques 5 years later (1995). Frozen plasma samples from baseline were available for a random sample of 152 men. Monolayers of endothelial cells cultured in micropore filter insets were pre treated with plasma, then normal human neutrophils were added to the endothelial cells and subsequent transmigration through the monolayers and micropore filters was measured. RESULTS: The endothelial monolayers were activated for transmigration of leukocytes more potently by plasma from participants with carotid artery plaques than participants without it. Increased endothelial activation with plasma at baseline was associated with the development of new atherosclerotic lesions during a period of 5 years. CONCLUSIONS: Plasma from individuals with prevalent atherosclerosis of the carotid arteries activates the endothelium for leukocyte transmigration, suggesting the presence of systemic pro inflammatory mediators. In an epidemiological survey, follow-up data on new lesion formation after 5 years indicated that plasma-mediated endothelium activation for interaction with leukocytes precedes the development of atherosclerotic lesions. PMID- 11006902 TI - Music perception and octave generalization in rhesus monkeys. AB - Two rhesus monkeys were tested for octave generalization in 8 experiments by transposing 6- and 7-note musical passages by an octave and requiring same or different judgments. The monkeys showed no octave generalization to random synthetic melodies, atonal melodies, or individual notes. They did show complete octave generalization to childhood songs (e.g., "Happy Birthday") and tonal melodies (from a tonality algorithm). Octave generalization was equally strong for 2-octave transpositions but not for 0.5- or 1.5-octave transpositions of childhood songs. These results combine to show that tonal melodies form musical gestalts for monkeys, as they do for humans, and retain their identity when transposed with whole octaves so that chroma (key) is preserved. This conclusion implicates similar transduction, storage, processing, and relational memory of musical passages in monkeys and humans and has implications for nature-nurture origins of music perception. PMID- 11006903 TI - Switching tasks and attention policies. AB - The determinants and costs of control were studied in 6 experiments examining the performance costs of changing stimulus dimension (digit value/number of elements) or attention strategies (speed/accuracy) on the first trial after task transition. Costs were compared for task shift and reconsideration only. Preparation ability was studied by presenting all transition information at the beginning of a 2-part block or only prior to each part. Results showed pronounced first-trial transition costs. Different factors were associated with stop-start and task-switching requirements. Transition costs were separate from those of basic task performance. Costs were sensitive to global control considerations and were larger for task dimension changes than for attention strategy shifts. Costs involving task dimension change, but not strategy shifts, were reduced with advanced preparation. These results are discussed in relation to contemporary models of control. A new distinction is proposed between activation and execution of control strategies. PMID- 11006904 TI - The task dependence of staged versus cascaded processing: an empirical and computational study of Stroop interference in speech production. AB - The authors investigated the on-line relationship between overt articulation and the central processes of speech production. In 2 experiments manipulating the timing of Stroop interference in color naming, the authors found that naming behavior can shift between exhibiting a staged or cascaded mode of processing, depending on task demands: An effect of Stroop interference on naming durations arose only when there was increased pressure for speeded responding. In a simple connectionist model of information processing applied to color naming, the authors accounted for the current results by manipulating a single parameter, termed "gain," modulating the rate of information accrual within the network. Results are discussed in relation to mechanisms of strategic control and the link between cognition and action. PMID- 11006905 TI - Language-dependent recall of autobiographical memories. AB - Two studies of autobiographical memory explored the hypothesis that memories become more accessible when the linguistic environment at retrieval matches the linguistic environment at encoding. In Experiment 1, Russian-English bilinguals were asked to recall specific life experiences in response to word prompts. The results supported the hypothesis of language-dependent recall: Participants retrieved more experiences from the Russian-speaking period of their lives when interviewed in Russian and more experiences from the English-speaking period of their lives when interviewed in English. In Experiment 2, the language of the interview was varied independently from the language of the word prompts. Both variables were found to influence autobiographical recall. These findings show that language at the time of retrieval, like other forms of context, plays a significant role in determining what will be remembered. PMID- 11006906 TI - Color categories are not universal: replications and new evidence from a stone age culture. AB - The authors sought to replicate and extend the work of E. Rosch Heider (1972) on the Dani with a comparable group from Papua, New Guinea, who speak Berinmo, which has 5 basic color terms. Naming and memory for highly saturated focal, non-focal, and low-saturation stimuli from around the color space were investigated. Recognition of desaturated colors was affected by color vocabulary. When response bias was controlled, there was no recognition advantage for focal stimuli. Paired associate learning also failed to show an advantage for focal stimuli. Categorical Perception effects for both English and Berinmo were found, but only at the boundaries of existing linguistic categories. It is concluded that possession of linguistic categories facilitates recognition and influences perceptual judgments. PMID- 11006907 TI - A sampling approach to biases in conditional probability judgments: beyond base rate neglect and statistical format. AB - Conditional probability judgments of rare events are often inflated. Early accounts assumed a general deficit in using statistical base rates. More recent approaches predict improvement when problems are presented in frequency format or refer to natural categories. The present theory focuses on sampling processes. Experiment 1 showed that a seeming advantage of frequency over probability formats is due to a confounded factor, the need to mentally transform stimulus samples. An information search paradigm was used in Experiment 2. When sampling by the predictor, the probability to be estimated, p(criterion/predictor), was conserved in the samples and judgments were quite accurate. However, when sampling by the criterion, the low base-rate event was strongly overrepresented, accounting for the entire bias. Judgments were quite sensitive to the sampled data, but failed to take sampling constraints into account, as shown in Experiments 3 and 4. PMID- 11006908 TI - [Hearing screening in newborn infants. Comparative studies and cost analysis with different instruments]. AB - BACKGROUND: The necessity of screening examinations in newborns today cannot be denied. Up to now, it has only been achieved in a few countries to introduce a general hearing screening for newborns. METHODS: We examined 100 newborns (200 ears) at their third day of life. All ears were evaluated with the TEOAE screening-device ECHOSCREEN. In addition, we carried out BERA screening examinations with the ALGO PORTABLE or with the EVOFLASH on 100 of these 200 ears. As a reference method, we utilized TEOAE-examinations with the ILO-88 for all ears. RESULTS: 4.5% of the 200 ears examined with the ECHOSCREEN were conspicuous due to accumulation of earwax problems. With the ALGO-system, none of the ears was conspicuous. Among the 100 ears examined with the EVOFLASH we found negative results with seven ears (4 children). These findings will be discussed in detail. Furthermore, we analyzed the costs for one-step- and two-step screenings. Costs for a one-step-screening of both ears would be DM 14.27 with Echoscreen, DM 44.69 with ALGO, DM 32.19 with Evoflash for one patient including material and personnel costs. Costs for a two-step-screening would be DM 16.28 with Echoscreen and ALGO, DM 15.72 with Echoscreen and Evoflash, calculated per child on the basis of 812,173 newborn children in the year 1997 in Germany. CONCLUSION: On the basis of all these results, we recommend a two-step-screening with TEOAE and BERA-devices. PMID- 11006909 TI - ["Vibrant Soundbridge" middle ear implant for auditory rehabilitation in sensory hearing loss. I. Clinical aspects, indications and initial results]. AB - BACKGROUND: The adequate therapy for patients suffering from a sensory hearing loss consists of fitting electronic hearing devices. Conventional hearing aids, however, present with significant inherent drawbacks such as insufficient amplification in the high frequency range, problems with the ear mold (feed back, occlusion, external otitis), or distortion of sound with an "unnatural" hearing impression. METHODS: The partially implantable middle ear device Vibrant Soundbridge provides a sound wave conversion into mechanical vibrations at the middle ear ossicles using the Floating Mass Transducer (FMT). The audiological advantages are due to a direct moving force to the perilymph via incus and stapes. The Vibrant Soundbridge system is indicated in patients with a medium to severe symmetrical sensory hearing loss and a normal middle ear. Candidates need previous experience with conventional hearing aids without satisfactory results. RESULTS: The eight operated patients report a "natural" quality of sound and speech, a better hearing perception at high frequencies and the absence of feed back phenomena. Audiological evaluation and questionnair results support the patients subjective hearing impression. CONCLUSIONS: The Vibrant Soundbridge improves hearing quality in patients with sensory hearing loss. The hearing implant is indicated in particular in patients that are unable to wear conventional hearing aids. PMID- 11006910 TI - [Studies of the histomorphology and function of the uvula]. AB - BACKGROUND: Alternations in pharyngeal structure and function are considered fundamental in the pathogenesis of snoring or obstructive sleep apnea (OSA). The physiological function of the uvula as a dynamic sealing of the nasopharynx prevents a "craniocaudal aspiration" during deglutition. The oropharyngeal soft tissues and the uvula are known to play an important role in affecting the oropharyngeal airflow resistance but studies about alterations in the histomorphological uvula structure are controversial. METHODS: We studied the histomorphological tissue composition of the uvula (midsagittal and transversal sections) in 142 patients who underwent uvulopalatopharyngoplasty (UPPP) or uvulopalatoplasty (UPP) for snoring and by autopsy in 30 normal subjects not known to have been snoring. Statistical comparisons were controlled for differences caused by age and body mass index. RESULTS: The uvula was found to be significantly longer in patients with snoring than in control subjects. Patients with snoring had a significantly greater percentage of fat content and connective tissue in combination with a muscle atrophy in the uvula than did normal subjects. CONCLUSIONS: The disturbance of the specific muscular composition and formation causes a reduction of muscular tonus and a loss of muscular contraction. The uvula is destabilized on the basis of a missing skeletal and cartilaginous brace. The uvula destabilization causes a narrowing of the pharyngeal airway that could lead to an increased oropharyngeal airflow resistance with an intensified passive uvula movement and vibration during mouth breathing. PMID- 11006911 TI - [Parotid involvement in cat scratch disease: a differential diagnosis with increased significance]. AB - BACKGROUND: Bartonella henselae is the causative agent of cat-scratch disease (CSD), an inflammatory infection of the lymph nodes. So far, only few cases of atypical manifestations in the head and neck, especially manifestations in the parotid gland have been reported. PATIENTS AND METHODS: Between January 1997 and June 1999 seven patients with manifestations of CSD in the parotid gland were observed at the ENT-department Freiburg. The positive diagnosis was confirmed serologically by an indirect immunofluorescence assay and by detection of Bartonella henselae-DNA with PCR-amplification and subsequent hybridization or sequencing. RESULTS: An intraglandular abscessed lymphadenitis was found in five patients, two of these cases were based on a Parinaud's oculoglandular syndrome. A diffuse affection of the parotid gland, initially misinterpreted as a parotid tumor, was seen in two patients. CONCLUSIONS: Antibiotic treatment is recommended in order to reduce the duration of the disease, in cases of pain and lymph node abscesses. In uncommon manifestations of CSD, the nosological assignment can be difficult on the basis of the heterogeneous symptomatic in the individual patient and the CSD diagnosis can only be confirmed by serology or PCR-based techniques. CSD should be considered in the differential diagnosis of all equivocal masses in the head and neck, even in unusual localisations like the parotid gland. PMID- 11006912 TI - [Second carcinomas in cancers of the mouth cavity, pharynx and larynx. Clinical, histopathologic and cell kinetic findings]. AB - BACKGROUND: The mortality because of malicious tumours of the oral cavity, of the pharynx and of the larynx has considerably increased in the Federal Republic of Germany during the last two decades. The prognosis not only shows a high ratio of recidivations and formation of metastases, but also a typical field canceration. There is a high rate of incidence of multiple primary, resp. secondary tumours that may occur synchronously or metachronously. Up to now we have learnt only little about the mechanisms concerning the development of tumours and their spread on the cellular molecular level. METHOD: The subject of the present retrospective study was the analysis of patients suffering from an epithelioma of the oral cavity, the pharynx and the larynx and the following secondary carcinomatas. The control group were patients suffering from primary tumours of the same location and subsequent recidivation or metachronous metastasis. By determination of the degree of malignity and keratinisation of the DNA ploidy, of the immunohistological expression of p53 and the immunohistological proliferation marker MIB1 conclusions had to be drawn with regard to the biological behaviour of tumours. RESULTS: The evaluation of the clinical parameters of the 122 patients on the whole revealed a clear increase of new disease as well as a shifting to the younger age within the last five years. Surprisingly, most common were secondary tumours in case of larynx carcinomatas. Patients suffering from secondary tumours show a bad prognosis, however, their maximum survival does not differ considerably from that of patients suffering from recidivations, resp. metastases. Because of the early diagnosis of the secondary carcinomatas the prognosis will depend on the primary carcinoma in most of the cases. The prognosis data indicate that independently from the fact whether there is a primary or a subsequent tumour, the therapy to be applied can only be a combined therapy consisting of operation and ray-therapy. The present examinations confirm reports with regard to disturbance of the p53 regulation that also play an important role in case of the head-neck area. The parallel analysis of MIB1 as a proliferation marker showed in case of the primary carcinomatas a correlation of positive p53-immunocolour with moderate and strong proliferation. Tumours in the hypopharynx and larynx showed a significantly smaller proliferation than that of the oral cavity and oropharynx. In case of the primary and secondary tumours the proliferation is more common in case of the G3-tumours. CONCLUSIONS: The in total modest differences between the primary and secondary carcinoma with regard to the DNA-ploidy; the proliferation and the p53-expression presumably originate in their formation within the scope of a so-called field canceration. Because of the field canceration supposed for the mucous membranes of the upper aerodigestive tract, the check-ups of these patients performed in regular intervals must not be limited in any case to the area of the primary tumour. They will have to consider the entire visible area of the upper respiratory and esophageal tract. PMID- 11006913 TI - [Lymph node metastases in the neck of unknown primary tumor]. AB - BACKGROUND: Cervical lymph nodes represent the most frequent manifestation of lymph node metastases of unknown primary. Nearly 3% of all malignant ENT-tumors are cervical lymph nodes metastases of unknown primary. This disease is a challenge for clinical working physician in diagnosis and therapy. MATERIAL AND METHOD: In a retrospective study we investigated 99 patients with the diagnosis cervical lymph node metastases of unknown primary, which were treated and observed in our department between 1975 and 1995. Within this group we observed the course of 83 patients completely. RESULTS AND CONCLUSIONS: The tumor dependent 5 year-survival-rate was 11%. This is very low, but similar to the literature. 40% of patients, that were operated on neck dissection with or without postoperative irradiation survived tumor-dependent 5 years. In 42 cases we could find a primary tumor. 14 of these primaries were located in the upper aero-digestive-tract, 28 in other regions of the body. The identification of the primary did not improve the prognosis of the patients. A good prognosis was associated with further occult primary, location of the lymph nodes in the upper or middle level of the neck or parotid region and a histology of squamous cell or undifferentiated carcinoma. Signs of poor prognosis were metastasis in the supraclavicular region, of adenocarcinoma and inoperability of the lymph node. The combination therapy of neck dissection and irradiation proved to be best. The extended field radiation of the complete upper aero-digestive-tract did not cause a improvement of tumor-dependent 5-year-survival. We discovered a primary in 5 of 27 patients in this group within the irradiated area. In conclusion extended field radiation must be discussed critically for patients with lymph node metastasis of unknown primary. PMID- 11006915 TI - [Interesting case no. 37. Jaffe-Lichtenstein fibrous dysplasia (osteodystrophia fibrosa)]. PMID- 11006914 TI - [Median sagittal mandibulotomy in head-neck tumors]. AB - BACKGROUND: Histologically proven radical resections are the goal in patients with head and neck cancer because of improved survival rates. The frequency of histologically radical resections using the median mandibulotomy and the morbidity of this approach for tumors of the oropharynx and the parapharyngeal space are described in a patients series. METHODS: The follow-up includes 16 consecutive patients who were operated on by a median mandibulotomy approach between 1995 and 1998. The oncological benefit (tumor free margins), complications and the functional results (ability of opening the mouth mastication, swallowing, speech, cosmesis and pain) were reviewed. RESULTS: In 15/16 cases a histological radical resection was achieved. 14 patients were irradiated postoperatively. In this group 3 patients had an osteoradionecrosis, one an osteomyelitis of the mandible. The functional results were worse in more advanced tumors. CONCLUSION: The indications for a median mandibulotomy are primarily T3 and T4 tumors of the oral cavity and oropharynx, rarely expansive benign parapharyngeal tumors. In most cases a histological radical resection is achieved even in advanced tumors, probably due to the wide exposure of the involved area. In contrast this approach is associated with a high morbidity. PMID- 11006916 TI - [Plastic surgery reconstructions in the neck area. Superficial neck soft tissue skin loss II]. PMID- 11006917 TI - Beyond the UKPDS: in search of the optimal strategy for the treatment of type 2 diabetes. PMID- 11006918 TI - Clinical effects of virgin olive oil polyphenols: between myth and reality. PMID- 11006919 TI - Evidence of postprandial absorption of olive oil phenols in humans. AB - BACKGROUND AND AIM: Olive oil phenols are potent antioxidants in vitro. If this were to be also demonstrated in vivo, it would help to explain the beneficial effects of this typical ingredient of the Mediterranean diet. This study was designed to determine the presence in lipoprotein fractions of two phenolic compounds peculiar to extra virgin olive oil, namely tyrosol and OH-tyrosol, and whether their absorption induces an antioxidant effect in vivo. METHODS AND RESULTS: Two trials were performed. In the first (Long-term), 14 healthy volunteers followed two diets, each for one month. The only difference between the diets was that the first supplied 50 g of extra virgin olive oil per day, where-as the second one supplied 50 g of refined olive oil with no simple phenols, as demonstrated by GC-MS analysis. There were no changes in LDL oxidizability and tyrosol and OH-tyrosol were not recovered in lipoproteins and plasma from fasting samples drawn at the end of each diet period. In the second study (Postprandial), eight healthy volunteers received an oral fat load consisting of 100 g of extra virgin olive oil. Blood was drawn at times 0', 30', 60', 120', 240', 360', and major plasma lipoprotein classes were separated. The concentration of tyrosol, OH-tyrosol and vitamin E was determined in lipoprotein fractions. Plasma antioxidant capacity was measured by a crocin-bleaching test and expressed as mM Trolox C equivalents. Tyrosol and OH-tyrosol were recovered in all lipoprotein fractions, except VLDL, with concentrations peaking between 60' and 120'. However, a very high variability in tyrosol and OH-tyrosol absorption was observed among subjects. Vitamin E content of LDL and HDL did not vary significantly throughout the study. Plasma antioxidant capacity increased significantly at time 120' (baseline 0.96 mM Trolox; 120' 1.19 mM Trolox; p = 0.02), and then returned almost to baseline values after 360' (1.1 mM Trolox). CONCLUSIONS: These findings suggest that phenolic compounds in olive oil are absorbed from the intestine, though not through a pathway dependent on chylomicron formation, and may exert a significant antioxidant effect in vivo, probably in the postprandial phase. PMID- 11006920 TI - The fatty acid composition of chylomicrons influences the rate of their lipolysis in vivo. AB - BACKGROUND AND AIMS: Previous work in our laboratory has shown that chylomicron triacylglycerol is lipolysed in vitro by lipoprotein lipase more rapidly when the particles are enriched with n-6 polyunsaturated as compared to saturated, monounsaturated or n-3 polyunsaturated fatty acids. It is possible, however, that this does not reflect the situation in vivo, where the active enzyme is bound to the vascular endothelium. The aim of the present study is to investigate the effect of the fatty acid composition of chylomicrons on their lipolysis in the rat in vivo. METHODS AND RESULTS: [3H]Oleate-labelled chylomicrons derived from palm, olive, corn or fish oil (enriched in saturated, monounsaturated, n-6 polyunsaturated and n-3 polyunsaturated fatty acids respectively) containing > 90% of the label in triacylglycerol were injected intravenously into functionally hepatectomised rats and blood samples were taken at time intervals up to 40 min. The radioactivity in serum triacylglycerol decreased significantly more rapidly when corn oil as compared to palm, olive or fish oil chylomicrons were used. Conversely, the radioactivity in serum free fatty acid derived from corn oil chylomicrons showed a faster increase than that derived from the other three types of particles. CONCLUSIONS: These results indicate that chylomicrons enriched with n-6 polyunsaturated fatty acids are converted to chylomicron remnants in vivo more rapidly than those enriched with saturated, monounsaturated or n-3 polyunsaturated fatty acids. This provides a partial explanation for the differential rate of removal from the blood of cholesterol carried in chylomicrons of different fatty acid composition demonstrated in previous work from this laboratory. PMID- 11006922 TI - Complement activation in hypercholesterolemia. AB - BACKGROUND AND AIM: Inflammatory and lipid factors share an important role in atherosclerosis. This study evaluates their relations in dyslipidemic subjects. METHODS AND RESULTS: We compared the complement system (serum hemolytic activity CH50, C3 and C4 fractions and terminal complex sC5b-9) in 30 hypercholesterolemic patients with elevated cholesterol and decreased HDL-cholesterol levels, 30 normolipemic patients with clinical atherosclerosis and 30 matched normal subjects. In addition we evaluated the circulating immune complexes containing cholesterol (chol-CIC) on the assumption that they might be important in complement activation, and the circulating levels of the adhesion molecule ICAM-1 (sICAM-1) as a sign of endhotelial dysfunction. We found a significant increase of sC5b-9 (but not of CH50 and C3, C4) in the hypercholesterolemics compared with the other groups. The plasma sC5b-9 level was inversely and significantly related to HDL-chol (regression analysis), whereas no direct significant relation was found between sC5b-9 and cholesterol. Chol-CIC were also significantly increased in this group. The atherosclerosis patients also presented a significant increase of sC5b-9. Lastly, both patient groups displayed a significant increase of sICAM 1. CONCLUSIONS: We suggest that complement activation in dyslipidemics may be induced by their increased immune complexes. However, the decrease of complement regulatory proteins carried by HDL is another important factor, while complement changes may be related to variations of other humoral and cell systems (endothelium, coagulative/fibrinolytic system), whose involvement is suggested in our study by the changes of sICAM-1. PMID- 11006921 TI - Dietary fatty acid composition influences the degree of human LDL oxidation, but has only minor effects on vascular tone in a bioassay system. AB - BACKGROUND AND AIM: Oxidized LDL has been detected in atherosclerotic vessels and presumed to be one of the major risk factors in atherosclerosis and cardiovascular diseases. The aims of the present study were to clarify whether the oxidation degree of LDL influences arterial tone and whether different long lasting dietary habits have effects on biological variables. METHODS AND RESULTS: The lag phase of LDL oxidation was shorter (117 +/- 6 min) in the fish diet group than in the vegetarian (153 +/- 5 min) or the control diet group (152 +/- 10 min). The rat mesenteric arterial rings, which were preincubated with LDL oxidized to 1-30%, from the vegetarian and the fish diet groups showed (p < 0.05) decreased NA-induced maximal contraction forces when compared to the control diet. The LDL oxidation degrees of 31-60% and 61-90% had no effect on NA- and KCl induced maximal contraction forces when compared to native LDL, nor were there differences between the diet groups. Endothelium-dependent and independent relaxation responses behaved similarly in all groups and were independent of the degree of oxidation. CONCLUSIONS: Dietary habits change the fatty acid composition of LDL, but have only minor effects on the vasoactive properties of oxidized LDL. PMID- 11006923 TI - Primary prevention of coronary heart disease: from controversy to consensus. AB - In recent years, interest has tended to focus on prevention of coronary events in high-risk groups, particularly those with established coronary heart disease. While this is understandable, it has led to a lack of emphasis on primary prevention. Yet it is only by means of primary or even pri-mordial prevention that a substantial reduction in coronary mortality on a population level will be achieved. This becomes clear when we consider that half of all persons who suffer a first myocardial infarction will die within the first month thereafter. Nevertheless, major progress has been made in primary prevention. Reliable risk algorithms have been constructed in Europe (PROCAM) and the U.S., and preliminary analyses on both sides of the Atlantic indicate that these algorithms can be useful applied to populations which are geographically and ethnically distinct from those in which they were derived. A notable trend in recent years is the increasing recognition of the metabolic syndrome with its key components of abdominal obesity, hypertriglyceridemia hypertension, low HDL-C, small, dense LDL, insulin resistance and hyperinsulinemia as being perhaps the most common and dangerous metabolic abnormality of all. Newer risk markers are being evaluated. The position of homocysteine remains unclear. Despite a strong association of elevated homocysteine with risk in case-control studies, prospective investigations have been less convincing. Evidence is beginning to accumulate from cross-sectional and prospective studies that markers of inflammation such as C-reactive peptide may improve our ability to predict risk of coronary events. While these data are encouraging, results of further studies must be awaited before the true place of these markers can be determined. The same can be said of many genetic markers of risk. Though a very large number of association studies have indicated links between a variety of genetic markers and coronary risk, these effects have tended to disappear after controlling for epigenetic and confounding factors and with increasing sample sizes. Finally, much attention is being devoted to non-invasive imaging of the coronary arteries. Such methods hold much promise as a screening test to exclude coronary stenosis in low-risk individuals. However, the measurement of calcium content of the arterial wall by EBCT has yet to prove its usefulness as a predictor of coronary events. PMID- 11006924 TI - Phyto-oestrogens and cardiovascular disease risk. AB - AIM: To present the currently available evidence on the cardiovascular benefits and risks associated with phyto-oestrogens. DATA-SYNTHESIS: Medline search from 1966-1999 updated with cross-check of references of papers with keywords such as phyto-oestrogens, isoflavones, lignans, genistein, daidzein, enterolactone, enterodiol, cardiovascular disease, cardiovascular disease risk factors. CONCLUSIONS: Phyto-oestrogens are plant chemicals divided into three main classes: isoflavones, coumestans, and lignans that display oestrogen-like activity due to their ability to bind to the oestrogen receptor. They are found in grains, beans, green vegetables, fruits, nuts, and grasses. Isoflavones are primarily found in soybeans and soy foods. For epidemiological studies of the relation between phyto-oestrogen intake and disease parameters, intake is estimated with several measures, such as biomarkers (concentrations in urine or blood) or dietary questionnaires, though the optimal method is not yet clear. Phyto-oestrogens are considered to act as selective oestrogen receptor modulators (SERM), exerting both oestrogen agonist and antagonist action. Supplementation with isolated soy protein containing the isoflavones genistein and daidzein reduces serum total and LDL-cholesterol and triglycerides in animals and in humans. Vascular reactivity might be improved by supplementation with isolated soy protein or isoflavones isolated from red clover. Studies on atherosclerosis in animals indicate a potential for risk reduction. Evidence in humans is still scanty. The little we know of the effects of regular dietary phyto-oestrogen intake comes from studies in which phyto-oestrogens were added to the usual diet. Most supplementation studies have been conducted with soy isoflavones, whereas the importance of lignans has not been determined, though they could be more important sources than isoflavones in Western populations. Research has been focused on risk factors. Studies of clinically manifest endpoints are urgently needed. PMID- 11006925 TI - The salt epidemic: old and new concerns. AB - Salt is the most suspected environmental factor which may influence arterial pressure. The recent demonstration that in the chimpanzee there is a strict dose response relationship between salt added to food and arterial pressure and that the large majority of these animals are "salt-sensitive" is solid proof that salt has a central importance in arterial hypertension. The behavioral complexity of mankind is such that the issue of salt sensitivity is very difficult to study in man. This is probably the main reason why all attempts to validate the arterial pressure response to tests of salt sensitivity based on changes in salt intake have produced negative or inconclusive results. However our inability to reliably identify salt-sensitive individuals in no way implies that the pressor effect of salt is evenly distributed in the population or that it is intrinsically so variable that the scientific efforts aimed at defining its nature are bound to fail. We envisage that in the near future the individual "salt-sensitive risk profile" will be calculated on the basis of new scientific knowledge about hypertension genes and environmental risk factors influencing the pressor response to salt. The link between target organ damage and salt is even stronger than that between arterial pressure and salt. Left ventricular mass is closely related to salt intake. In the kidney there is evidence that high salt intake induces hyperfiltration and raises glomerular pressure. Of note the damaging potential of salt to the heart and the kidney seems to be largely independent of the ongoing arterial pressure response. A widespread reduction in salt intake at population level might in theory produce major health benefits but it seems unlikely that major public health actions will be undertaken in the near future. PMID- 11006926 TI - [The pathologist as a discover of patterns. Perspectives in disease morphology]. PMID- 11006927 TI - [History of histopathology]. PMID- 11006928 TI - [Pituitary necrosis. Decreasing incidence due to modern intensive therapy?]. AB - Pituitary necroses develop from the interruption of circulation in the pituitary stalk, especially due to shock, trauma, etc. We studied the incidence, extent, and outcome of necroses in postmortem series from 1991 to 1998 and compared these with the findings in our own study in the 1970s. Necroses were classified into four degrees based on the proportion of anterior pituitary volume: grade I, up to 5%; grade II, 5-10%; grade III, 10-50%; grade IV, more than 50%. The data show a decreased incidence of necrosis, from 7.7% in the 1970s to 3.1% in the 1990s. Scars, indicating former necroses, decreased in incidence from 11.4% to 3.6%. The changes in the four grades of necrosis were: a decrease in grade I from 60% to 41%, a decrease in grade II from 24% to 22%, an increase in grade III from 9% to 17%, and an increase in IV from 8% to 20%. These figures reveal that pituitary necroses became less frequent but more extensive. Necroses of grade IV are generally caused by more than a single cause. The decreased incidence appears to be the consequence of progress in intensive care over the past 20 years, as patients can now survive severe shock longer. The increased frequency of intensive necroses is probably due to the longer period for greater extent of necrosis to develop during intensive care. PMID- 11006929 TI - [Diagnostic importance of flow cytometry in staging malignant lymphoma]. AB - Bone marrow biopsy plays an important role in the clinical diagnosis of malignant lymphoma. Further diagnostic methods need to be established to increase accuracy in the light of advances over recent years in the immunophenotypical characterization of discrete lymphatic bone marrow lesions and the continuing difficulty in classifying them. This study compared the diagnostic value of flow cytometry to that of conventional bone marrow biopsy in 156 patients with 191 marrow biopsy specimens or bone marrow aspirates. The most important findings were that up to one-third of lymphomas could not be diagnosed by flow cytometry, and that the degree of infiltration was estimated as less than two-thirds. However, flow cytometric results were more satisfactory in acute lymphoblastic leukemia, lymphoblastic lymphoma, hairy cell leukemia, and chronic lymphocytic leukemia. In summary, flow cytometry has a complementary role in the staging of lymphoma but cannot fully replace conventional trephine biopsy. PMID- 11006930 TI - [Spontaneous complete rupture of a thrombotic aneurysm of the tibiofibular trunk]. AB - True arterial aneurysms of the infrapopliteal vessels are a rare finding. During surgical treatment of convergent aneurysms of the popliteal artery and the tibiofibular trunk we discovered a complete spontaneous rupture of the thrombotic aneurysm of the right tibiofibular trunk. We describe the special features of diagnosis, histology, and the surgical treatment of aneurysms of the crural arteries. We recommend that even asymptomatic aneurysms be electively operated on to prevent further complications. PMID- 11006931 TI - [Acardius or "twin-reversed arterial prefusion" sequence. Report of four cases and review of current therapeutic possibilities]. AB - We describe four cases of acardius (one of hemiacardius, two of holoacardius acephalus, and one of the holacardius amorphus) and present the current range of therapeutic possibilities: control of amniotic fluid by repeated amniocentesis or indomethacin therapy, administration of digoxin to the mother, selective preterm delivery of the acardiac twin by hysterotomy, interruption of the anastomoses by injection of thrombogenic coils, sclerosing agents, umbilical cord ligation, fetoscopic laser coagulation, and thermocoagulation. PMID- 11006932 TI - [Serous cystadenofibroma of the epiploic appendix. A tumor of the secondary mullerian system: case report and review of the literature]. AB - We present a case of serous cystadenofibroma 2 cm in diameter in the epiploic appendix of the sigmoid as incidental finding in a 72-year-old patient who underwent hysterectomy and oophorectomy for endometrial carcinoma. The tumor showed the same histology as analogous tumors of the ovary and was associated with endosalpingiosis. Further findings were large adhesions between the epiploic appendices of the sigmoid and the parietal peritoneum and atypical cells in the peritoneal washings. Both may be explained by occult peritoneal endosalpingiosis. The histogenesis, histology, and locations of extraovarian mullerian tumors are reviewed. PMID- 11006933 TI - [Sudden cardiac death during sports due to coronariitis]. AB - A case of sudden and unexpected death of a 29-year-old man during a basketball game is reported. Six years before death a transient ischemic attack and a membranous glomerulonephritis were diagnosed. The autopsy revealed as cause of death to be a severe lymphoplasmocytic coronariitis and, in addition, a small single focus of a lymphoplasmocytic aortitis, indicating a systemic disease. A suspected syphilitic vasculitis could not be confirmed. In the literature there are few cases of similar coronary arteriitis are described, such as Takayasu's type of arteriitis in men of young and middle age. PMID- 11006934 TI - [Endemic syphilis in the recognized relics..... Letter to Der Pathologe (5/99) 20:292-296]. PMID- 11006935 TI - [In Process Citation] PMID- 11006936 TI - [Bone marrow involvement in hematologic neoplasms and solid tumors]. AB - Bone marrow involvement is often observed in patients with hematological malignancies and solid tumors. Sensitive immunological and molecular biological methods allow the detection of isolated transformed cells in bone marrow samples. Like normal hematopoietic cells, tumor cells interact with bone marrow stroma through adhesion molecules. Since adhesion molecules play an essential role in the invasion and proliferation of malignant cells, new therapeutic approaches involving the inhibition of cell-cell and cell-matrix interactions are conceivable. PMID- 11006937 TI - [Micrometastatic cells in the bone marrow of patients with breast carcinoma]. AB - BACKGROUND: The immunocytological detection of disseminated epithelial cells in bone marrow in patients with breast cancer has been performed at many hospitals and institutes since the early 1980s. Despite numerous publications in this field, it has not been possible to standardize the method and establish the ideal antibody, either nationally or internationally. Molecular biological methods using PCR technology could extend the diagnostic spectrum. However, one of the major problems in breast cancer is the lack of a disease-specific marker gene. As a result, immunocytology is still the standard procedure for tumour cell detection. METHODS: The detection of disseminated single cells in bone marrow in primary breast cancer (also known as minimal residual disease) is a new prognostic factor for disease-free and overall survival. This has been demonstrated in three large (N > 300) groups and several small to medium groups (N = 50-300). As a marker of dissemination in a target organ for metastasis this prognostic factor corresponds much more closely to the tendency of breast cancer to early haematogenic spread. Tumour cell detection may predict the course of the disease better than the axillary lymph node status. Bone marrow aspiration and detection of disseminated cells might replace lymph node dissection, at least in those patients with small tumours and no clinical signs of lymph node involvement. This strategy will soon be investigated in appropriate studies. Another possible clinical use might be deciding on whether or not to give adjuvant systemic therapy to node-negative patients. Patients with positive tumour cell detection are at a higher risk of subsequent metastasis, even if the axillary nodes are histologically normal. APPLICATION OF METHODS: The immunohistological or molecular biological detection of tumour cells in axillary lymph nodes might also be very useful, now that is has been shown that a considerable subset of patients determined to be node-negative by means of conventional methods, are positive according to these new techniques. These methods could be a useful supplement to sentinel node biopsy. A further potential use of this method is in monitoring therapy with new treatment modalities such as gene therapy and immunotherapy. Repeated bone marrow aspiration can provide information on the success of therapy in minimal residual disease (cytoreduction). Immunocytochemical investigation of individual cells may be useful in studying the pathogenesis of metastasis, in particular in the skeleton. Phenotyping of cells might allow statements to be made in the metastatic potential of cells and the question of cell dormancy. It remains to be hoped that this aspect of minimal residual disease will be granted more attention in future. PMID- 11006938 TI - [The normal bone marrow and its variations in MRT]. AB - Physiology and age-dependent changes of human bone marrow are described. The resulting normal distribution patterns of active and inactive bone marrow including the various contrasts on different MR sequences are discussed. PMID- 11006939 TI - [Proton-MR spectroscopy of the spinal bone marrow. An analysis of physiological signal behavior]. AB - BACKGROUND: Magnetic resonance imaging has shown to be a sensitive method for diagnostics of the red bone marrow, the composition of which changes physiologically and during pathological processes. However, the interpretation of MRI in patients with disorders of the red bone marrow is very difficult. The aim of this study was the characterization of the proton spectrum of healthy bone marrow and its age- and sex-dependent changes to obtain a data basis for measurements in patients. METHODS: 154 healthy volunteers have been examined. After imaging, a spectroscopic measurement was performed to determine the relative intensities of fat and water, and their respective T2 times. RESULTS: While T2 (water: 46.9 ms and fat: 75.4 ms) does not depend on age or sex, the relative signal intensity of fat increases by about 6% per decade. In the age groups between 31 and 50 years it diverges significantly between men (43.5%) and woman (32.5%) (p < or = 0.01, Mann-Whitney-Test. CONCLUSIONS: Proton spectroscopy can increase the reliability of diagnosis by offering information on composition of the marrow. The analysis of spectroscopic measurements requires exact knowledge about normal physiological values. PMID- 11006940 TI - [The MR characterization of the composition of the hematopoietic bone marrow. The findings in generalized neoplasms and the monitoring of therapy]. AB - PURPOSE: Methodological work was performed in the field of magnetic resonance imaging (MRI) and spectroscopy (MRS) in order to develop suitable tools for non invasive characterization of hematopoietic bone marrow. The methods were applied for the assessment of normal values in healthy persons and to examine patients with generalized hematological diseases or to monitor effects of therapies influencing the composition of bone marrow. METHODS: Besides standard techniques of MRI as T1- or T2-weighted methods, chemical shift techniques for selective visualization of water or lipid components were applied. The method of magnetization transfer (MT) contrast was used with the intention to differentiate between multiple water containing tissue compartments (intra- vs. extracellular space). A further approach was the determination of the magnetic field distribution within spongy bone marrow. Besides investigations in healthy volunteers, prospective clinical studies were carried out in patients suffering from acute leukemia during their initial treatment and in patients who underwent high-dose therapy with following peripheral blood stem cell transplantation (PBSCT). RESULTS: Especially MR techniques for selective imaging of water of fat signals and proton spectroscopy yielded a high sensitivity to primarily pathological or therapeutically induced changes of hematopoietic bone marrow. Application of MT allowed an improved differentiation of the tissue compartments under PBSCT, which might result in temporary edema. Storage of hemosiderin in bone marrow after blood transfusions and simultaneous hematopoietic insufficiency could be revealed by methods sensitive to magnetic field inhomogeneities. CONCLUSIONS: Methods of MRI and MRS allow to non-invasively characterize hematopoietic bone marrow in the course of hematological diseases and during therapy. Marked changes in the composition of hematological bone marrow are detectable for extensive marrow areas. The prognostic relevance of the findings has to be evaluated in future follow-up studies. PMID- 11006941 TI - [The diagnostic problems in magnetic resonance tomography of the bone marrow in patients with malignomas under G-CSF therapy]. AB - AIM: To study the effect of G-CSF therapy directly by MRI and 1H MRS in the lumbar and femoral bone marrow and differentiate between malignant bone marrow infiltration (MBMI) and reconversion of red marrow. METHODS: Thirteen patients could be examined twice, before and during G-CSF medication and another six only during treatment. T1 weighted spin-echo and opposed-phase gradient-echo images as well as the spectroscopic data (T2 values, water content) were analysed. RESULTS: After G-CSF a pathologic bone marrow signal intensity was seen in 8/13 (lumbar) and 11/13 (femoral) patients respectively. The majority of the signal alterations were diffuse (6 and 8), the minority focal (2 and 3). If a patient was successfully stimulated, a significant increase in water content occurred (21% lumbar, 34% femoral). T2 values did not change significantly, nor did they correlate with the stimulation success. CONCLUSIONS: MR tomography and spectroscopy are suitable to detect lumbar and femoral bone marrow stimulation by G-CSF quantitatively and qualitatively. The changes may simulate MBMI. The adequate judgement of G-CSF treated bone marrow without pretherapeutic images is not possible. PMID- 11006942 TI - [The diagnosis of plasmocytoma with MRT]. AB - BACKGROUND: In multiple myeloma 5 different infiltration patterns can be differentiated: 1. normal appearance of bone marrow, 2. focal involvement, 3. homogeneous diffuse infiltration, 4. combined diffuse and focal infiltration, 5. "salt- and pepper" pattern with inhomogeneous bone marrow with interposition of fat islands. METHODS: For the fast and total acquisition of all patterns a combination of a T1-weighted spin echo sequence and a fat suppression technique is superior. The focal involvement is clearly demonstrated as areas of high signal intensity on e.g. STIR images. Diffuse involvement can be quantified objectively by calculation of the percentage of signal intensity increase after contrast material injection. MRI is superior to X-ray in focal and diffuse involvement. With ultrafast sequences a "screening" of the whole red bone marrow as for myeloma infiltration is possible. PROGNOSIS: In prognosis studies diffuse infiltration is inferior to focal involvement. Patients without bone marrow infiltration have a significantly longer survival than patients with bone marrow infiltration in MRI at the time of diagnosis. However, even patients in stage one of disease (Durie and Salmon) and negative X-ray films can show bone marrow infiltration in MRI. Those patients often show an early disease progression. Good response to therapy in focal involvement are: reduction of signal intensity on T2 weighted spin echo images, lack or rim-like enhancement after contrast material injection or even a normalisation of bone marrow signal. In case of diffuse involvement a partly patchy reconversion to fatty marrow can be seen. PMID- 11006943 TI - [Functional magnetic resonance tomography in the diagnosis and therapy monitoring in multiple myeloma]. AB - AIM OF THE STUDY: Investigation of the quantitative microcirculation parameters amplitude A and exchange rate constant k21 determined by contrast-enhanced dynamic magnetic resonance imaging (d-MRI) in multiple myeloma (MM). METHODS: d MRT of lumbar spine and right spina iliaca superior posterior of 16 controls (ctr) and 35 patients with active MM. Generation of colour-coded images of microcirculation parameters superimposed onto static MRI images. RESULTS: Amplitude A and k21 parameters were significantly increased in patients with MM and down modulated by therapy in 7 of 8 MM cases in a follow-up investigation [p < 0.01; median Actr = 0.2 (0.09-0.4); median AMM = 0.93 (0.2-1.52); median k21ctr = 0.09 min-1 (0.03-0.9); median k21MM = 4.57 min-1 (0.21-23.8)]. Thirteen patients revealed a "diffuse" and 22 a "focal" pattern of distribution of microcirculation parameters. Bone marrow biopsies in 8 cases revealed an correlation between bone marrow plasma cell infiltration and increased microcirculation parameters. CONCLUSION: Identification of microcirculation changes by d-MRI is a novel imaging technique for the detection and monitoring of MM bone lesions. PMID- 11006945 TI - [Bone manifestations in non-Hodgkin's lymphoma in childhood and adolescence]. AB - PURPOSE: Skeletal manifestation of Non-Hodgkin's lymphoma is rare in pediatric patients. Objective of the study was to determine imaging features, before and after treatment, and to correlate these features with clinical outcome. METHODS: A retrospective analysis of 1246 patients from two therapy studies (NHL-BMF-90 and 95) was performed. Imaging studies of 63 patients with bone involvement of lymphoma were reevaluated. RESULTS: Incidence of initial bone involvement in Non Hodgkin's lymphoma was 6.8%. Distribution was best assessed by bone scan, MRI revealed larger areas of marrow involvement and detected additional lesions. Sites of predilection were long bones of the lower extremities with epiphyseal involvement in 39%. Residual signal alterations in MRI after successful therapy remained in 71%. Osteonecrosis after therapy was a common finding. Clinical outcome war not correlated to the presence of bone involvement. CONCLUSIONS: Since clinical outcome is not effected by bone involvement in childhood NHL, value of screening may be limited. Knowledge of imaging characteristics is mandatory for initial evaluation of primary osseous lymphomas and symptomatic lesions as well as for therapy controls. PMID- 11006944 TI - [The value of diffusion-weighted MRT in assessing the bone marrow changes in vertebral metastases]. AB - AIM OF THE STUDY: The aim of the study was the evaluation of the diffusion coefficient (ADC) of vertebral metastasis and regular vertebral bodies with diffusion weighted MRI (DWI). DWI evaluates the tissue-specific molecular diffusion of protons. In tissues with high cell densities (neoplasm) a decreased ADC can be expected due to restricted diffusion according to an exaggerated amount of intra- and intercellular membranes (i.e. diffusion barriers). METHODS: In 5 breast cancer patients the ADC of both known vertebral metastases and of adjacent regular vertebral bodies were measured with DWI (1.0 T; Phased-Array Body-Coil; b: 880 and 440 s/mm2). RESULTS: The ADC of regular vertebral bodies (1.3 +/- 0.23 x 10(-3) s/mm2) was significantly (p < or = 0.0002) higher than in vertebral metastases (0.39 +/- 0.11 x 10(-3) s/mm2). CONCLUSIONS: These data demonstrate that the ADC can be reliably measured in vertebral bodies. The quantitative evaluation of the ADC in vertebral bodies seems to be an objective and comparable parameter for differentiating malign from benign vertebral tissue. PMID- 11006946 TI - [Ischemia of the lower extremity due to a persistent a. ischiadica--a possible interventional therapeutic approach]. AB - The sciatic artery is a rarely seen variant of the vessels of the lower limb. Anatomically it is the prolongation of the inferior gluteal artery and accompanies the sciatic nerve from the sciatic foramen to the knee. More than other vessels, the sciatic artery shows vasculopathies like diffuse dilatation or aneurysms. This report describes an interesting case of embolic acute ischemia, caused by a persistent sciatic artery with an aneurysm. We treated the ischemia with intra-a. lysis and bridged the aneurysm with an covered stent. PMID- 11006947 TI - [Sellink is not "by" Sellink. A letter on the work of S. Feuerbach and J. Scholmerich, Radiologe (2000) 40: 324-38]. PMID- 11006948 TI - [Intraspinal, epidural space-occupying lesion]. PMID- 11006949 TI - [Interventions in hemodialysis shunts]. PMID- 11006950 TI - NMDA blockade attenuates caspase-3 activation and DNA fragmentation after neonatal hypoxia-ischemia. AB - The aim was to study the effects of an NMDA receptor antagonist on caspase-3 activation and DNA fragmentation after hypoxia-ischemia (HI) in 7-day-old rats. Animals were treated with vehicle or MK-801 (0.5 mg/kg) directly after HI and sacrificed 8, 24 or 72h later. MK-801 reduced injury (by 53%), cells positive for active caspase-3 (by 39%) and DNA fragmentation (by 79%) in the cerebral cortex. Furthermore, MK-801 significantly decreased caspase-3 activity, and Western blots revealed a tendency towards decreased proteolytic cleavage of the caspase-3 proform. The data imply that NMDA receptors are involved in the activation of apoptotic processes in the immature brain after HI. PMID- 11006951 TI - Growth regulation of astrocytes and C6 cells by TGFbeta1: correlation with gap junctions. AB - Transforming growth factor (TGF) beta1 enhanced in vitro [3H]thymidine incorporation into C6 cells and reduced that of astrocytes in the presence of a high serum concentration. It concomitantly raised the gap junction intercellular communication (GJIC) in normal astrocytes but reduced the coupling of C6 cells, and respectively increased or decreased the proportion of P2-phosphorylated connexin (Cx) 43 isoform in these cells. Finally, octanol, which inhibited GJIC in both cell types, increased the thymidine incorporation in C6 cells, but neither altered the proliferation of astrocytes nor their response to TGFbeta1. These data indicate that an inhibition of gap junction intercellular communication, due to an altered phosphorylation of connexin 43, may contribute to the proliferative response of C6 glioblastoma cells to TGFbeta1. PMID- 11006952 TI - Visually induced activity in human frontal motor areas during simple visuomotor performance. AB - Visuomotor tasks elicit neuronal activity in primate motor areas at relatively short latencies. Although this early activity embodies features of visual responses (short latency, stimulus-dependency), its sensory nature has been questioned. We investigated neural correlates of visuomotor performance in human motor areas using scalp and intracranial event-related potential measures. A simple visuomanual reaction-time task evoked early potentials at 133-145 ms post stimulus which occurred much earlier than the motor potentials of the same region. The amplitude of the early potentials covaried with stimulus location and was independent of parameters of the motor response. Because of their timing, stimulus-dependency and characteristics of our behavioral task, the early potentials are suggested to reflect neuronal responses of sensory nature rather than processing related to pure motor aspects of the task. PMID- 11006953 TI - Effects of serotonergic 5-HT1A and 5-HT1B ligands on ventral pallidal neuronal activity. AB - To clarify the role of the 5-HT system in limbic outputs, the present study compared the effects of the 5-HT1A agonist 8-OH-DPAT and the 5-HT1B agonist CP 94253 with the non-selective 5-HT agonist TFMPP on the firing rate of ventral pallidal (VP) neurons recorded in chloral hydrate-anesthetized rats. 8-OH-DPAT (0.25-256 microg/kg i.v.) dose-dependently enhanced (9/26 neurons) or suppressed (8/26) activity, and the 5-HT1A antagonist (+)WAY-100135 often attenuated these responses. TFMPP (0.011-1.453 mg/kg i.v.) dose-dependently reduced the firing rate of 7/8 VP neurons tested. In contrast, CP-94253 (0.013-12.8 mg/kg i.v.) had little or no effect. In sum, these data suggest that the 5-HT1A receptor appears to be particularly important in influencing limbic outputs mediated via the VP. PMID- 11006954 TI - Expression of intercellular adhesion molecule-1 in C6 glioma cells is up regulated by thyroid hormone. AB - ICAM-1 is a cell surface adhesion glycoprotein playing an essential role in inflammatory responses. We have investigated the effects of the thyroid hormone T3 on the expression of the ICAM-1 gene in C6 glioma cells. In these cells, T3 stimulated the ICAM-1 protein expression significantly after 24 h of treatment. The induction of ICAM-1 by cytokines such as interleukin 1beta or tumour necrosis factor TNF as well as by lipopolysaccharide or T3 can be suppressed by the two anti-inflammatory compounds dexamethasone and parthenolide. The C6 glioma cell line could then be a useful model for studying the effect of T3 hormone on the expression of specific genes in glial cells, especially genes involved in lymphocyte-glial cell interactions. PMID- 11006955 TI - PET shows that odors are processed both ipsilaterally and contralaterally to the stimulated nostril. AB - The olfactory nerve is the only cranial nerve with established ipsilateral primary cerebral anatomical projections. Whether these projections correspond to the functional pathways for monorhinal processing of odor perception is, however, unknown. We therefore studied cerebral blood flow (rCBF) with [15O]butanol-PET in 18 healthy females during monorhinal smelling of single odors (OS) and odorless air (AS). Compared with AS, OS activated right amygdala and piriform cortex (confluent cluster), right orbitofrontal cortex, left insula, right thalamus, and anterior cingulate. A post hoc analysis showed that the first three regions were activated independently of the stimulated side, but that right orbitofrontal rCBF was higher during the right nostril stimulations. Left insula was activated mainly by the right nostril stimuli, and right thalamus by the left nostril stimuli. Odors seem to be processed both ipsi and contralaterally, with a right hemisphere preponderance irrespective of the stimulated nostril. PMID- 11006956 TI - A study of the apparent diffusion coefficient of grey and white matter in human ischaemic stroke. AB - We examined whether there was any difference in the value, and temporal evolution, of the apparent diffusion tensor trace (ADC) in acute and subacute grey and white matter lesions. Thirty-seven patients underwent diffusion imaging once (up to 3 days), 15 patients were scanned twice (up to 7 days), and seven patients were scanned three times (up to 14 days) after stroke. Values of the ratio of ischaemic to contralateral ADC (ADCr) were reduced on average by 30% (p<0.001) in the whole hyperintense region up to 7 days post-ictus. No difference was seen between ADCr values of grey and white matter in individual subjects within the patient groups scanned up to 7 days. However, in the subgroup of patients scanned beyond 7 days, ADCr for grey matter rose significantly (p=0.02) from ADCr approximately 0.7 (< 7 days) to 0.95 (> or = 10 days). This increase did not occur in white matter whose ADCr remained fairly constant (ADCr approximately 0.7) over the time course of the study. PMID- 11006957 TI - Causal influences in primate cerebral cortex during visual pattern discrimination. AB - Anatomical studies of the visual cortex demonstrate the existence of feedforward, feedback and lateral pathways among multiple cortical areas. Yet relatively little evidence has previously been available to show the causal influences of these areas on one another during visual information processing. We simultaneously recorded event-related local field potentials (LFPs) from surface to-depth bipolar electrodes at six sites in the ventral region of the right hemisphere visual cortex in a highly trained macaque monkey during performance of a visual pattern discrimination task. Applying a new statistical measure, the short-time directed transfer function (STDTF), to the LFP data set, we charted the changing strength and direction of causal influence between these cortical sites on a fraction-of-a-second time scale. We present results showing, for the first time, the dynamics of distinct feedforward, feedback and lateral influences in the ventral portion of the primate visual cortex during visual pattern processing. PMID- 11006958 TI - Global and local processing of visual patterns in macaque monkeys. AB - Human subjects generally perceive the global form of hierarchically organized visual patterns faster than the local form. To test whether macaque monkeys show a similar precedence for global processing, two monkeys were trained to discriminate either the global or local form of hierarchical visual patterns. The response time to discriminate the global form was shorter than that to discriminate the local form. Consistent patterns, in which the global and local forms were identical, were discriminated faster than inconsistent ones, in which the two forms were different from each other, both in discrimination of the global form and of the local form. Similar results were obtained in two human subjects who were subjected to the identical tests. The results suggest that, both in monkeys and humans, the global form is processed faster than the local form with a temporal overlap which allows bidirectional interactions between the two processes. PMID- 11006960 TI - Simultaneous bilateral mismatch response to right- but not leftward sound lateralization. AB - Magnetoencephalography (MEG) was used to compare mismatch responses between hemispheres to changes in sound-source direction. Sixteen adults listened passively to two types of complex non-language sounds presented in separate blocks with midline standards and right- and left-lateralized deviants. Mismatch dipole amplitudes were larger contra- than ipsilaterally to the deviants. Both hemispheres processed right deviants simultaneously, whereas to left deviants, the left dipole peaked 20 ms later than the right dipole. A second experiment using the same standards but midline spectral deviants showed no interhemispheric differences. Here mismatch latencies were about 60 ms longer than in the location mismatch experiment. This suggested both fast, contralaterally dominant location mismatch responses and facilitated detection of auditory spatial deviance in the right hemifield. PMID- 11006959 TI - Involvement of the sigma1 receptor in the cocaine-induced conditioned place preference. AB - The sigma1 (sigma1) receptor constitutes a particular target of cocaine believed to be involved in some of its behavioral effects. In the present study, its involvement in the rewarding effect of cocaine was examined using the conditioned place preference (CPP) procedure. CPP was induced in C57Bl/6 mice injected repeatedly with cocaine (20 mg/kg, i.p.). The selective sigma1 receptor antagonists NE-100 and BD1047 (1-10 mg/kg, i.p.) significantly attenuated or blocked the cocaine-induced CPP. Animals treated centrally with a sigma1 receptor antisense oligodeoxynucleotide failed to develop cocaine-induced CPP, unlike mismatch controls. The sigma1 receptor thus appears to be critically involved in the development of the cocaine-induced CPP and, in consequence, may constitute a promising approach to blocking cocaine reward. PMID- 11006961 TI - Auditory cortex evoked magnetic fields and lateralization of speech processing. AB - Potential use of different auditory evoked brain responses for determining cerebral lateralization of speech function was evaluated. Cortical magnetic fields elicited by plosive syllables or complex non-speech sounds analogous to them were recorded with 122-channel magnetometer. We estimated parameters of magnetic P1, N1 and P2 responses to both stimuli in the two hemispheres and found no hemispheric asymmetry for any of the responses. No correlation between the right-ear advantage, determined with dichotic listening test, and any of asymmetry indexes, calculated for the speech-elicited responses, was observed. These results suggest that P1, N1 and P2 responses to speech signals do not indicate lateralization of speech function in the brain. The results are discussed in relation to previous studies suggesting that the mismatch negativity (MMN) seems to be the only early auditory cortex response sensitive to the lateralization of speech function. PMID- 11006962 TI - Mossy fiber sprouting is dissociated from kindling of generalized seizures in the guinea-pig. AB - Controversy surrounds whether aberrant mossy fiber sprouting in the hippocampus is necessary for the establishment of seizure states. We investigated the association between mossy fiber sprouting and kindling in guinea-pigs, using either single-site or alternate-site stimulation. Kindling with single-site amygdaloid stimulation did not induce significant sprouting, despite the development of partial seizures. In contrast, single-site septal and alternating amygdaloid-septal stimulation produced moderate but significant sprouting in about 60% of animals that failed to develop stage 5 generalized seizures. Since the magnitude of sprouting was similar despite striking differences in the intensity of seizures that developed, we conclude that mossy sprouting is not causally associated with seizure development. PMID- 11006963 TI - Rapidly turned over protein maintains metabotropic synaptic transmission in Purkinje cells. AB - It has generally been thought that protein synthesis is required for relatively slow cellular processes such as synaptogenesis and synaptic plasticity. In this study on rat cerebellar slices, we found that metabotropic glutamate receptor mediated synaptic transmission to cerebellar Purkinje cells was quickly and persistently depressed by brief (5 min) applications of translational inhibitors, which were confirmed to induce quick and persistent depression of protein synthesis in cerebellar tissues. Brief applications of transcriptional inhibitors also depressed metabotropic synaptic transmission, but progressively over 1 h, presumably due to depletion of mRNAs in the dendrites. Results of this study indicate the presence of a unique protein(s) that is dynamically involved in metabotropic synaptic transmission. PMID- 11006964 TI - Language functional neuro-imaging changes following focal left thalamic infarction. AB - The involvement of the left thalamus in language function has been largely demonstrated through the effects of thalamic lesion and/or stimulation upon language. However, the pathophysiological mechanisms underlying thalamic aphasias remain a matter of debate. We report here on changes in the pattern of brain activity in auditory word processing, verb generation and visual lexical decision in a patient who sustained left thalamic infarct following the surgery of intractable mesio-temporal epilepsy with left hippocampal sclerosis. To some extent, our findings exemplify the way the left thalamus brings on line the cortical networks involved in language processing, suggested in the term 'selective engagement'. PMID- 11006965 TI - Cytoplasmic residues influence the voltage-dependence of the gating of human K+ channels. AB - Shaker type potassium channels are strongly voltage dependent and potassium selective. Kv1.4 channels from a variety of sources exhibit a much shallower voltage-dependence of activation than other members of the family. We have made a chimeric construct consisting of the N-terminal chain of hKv1.1 spliced onto the transmembrane portion of hKv1.4 (IN/4). When expressed in Xenopus oocytes, the chimeric channel exhibits a voltage dependence that is similar to hKv1.1 although the voltage sensing and transduction machinery presumably reside in the transmembrane portion of the channel. Loss of the N-terminal ball and chain from hKv1.4 is not responsible for this as a truncation construct, starting close to the splice junction, has the same voltage-dependence as full length hKv1.4. We suggest that residues from the N-terminal chain of hKv1.1 interact with the machinery that transduces movement of the voltage sensor into channel opening. If so, this chimeric construct could provide a handle to the identification of elements of this transduction machinery. PMID- 11006966 TI - A systematic representation of interaural intensity differences in the auditory cortex of the pallid bat. AB - The current model of cortical processing of auditory spatial information is based on an orthogonal representation of frequency and binaural response properties, but how this arrangement leads to representation of space in the auditory cortex is unclear. This study describes the first evidence of a cortical substrate for the systematic representation of space in a region of primary auditory cortex of the pallid bat that subserves passive sound localization. The organizational feature of this region is a systematic shift in sensitivity to interaural intensity differences across the cortical surface, suggesting a topographic representation of horizontal space based on the distribution of activity within the neuron population. PMID- 11006967 TI - Chronic repetitive transcranial magnetic stimulation induces subsensitivity of presynaptic serotonergic autoreceptor activity in rat brain. AB - Repetitive transcranial magnetic stimulation (rTMS) is a novel procedure which has proven effective in the treatment of major depression. We administered rTMS chronically to rats in order to determine whether this procedure affected serotonergic neurotransmission in the prefrontal cortex. Basal 5-HT levels, and the effects of challenges with the 5-HT1A receptor agonist 8-OH-DPAT and the 5 HT1B antagonist GR 127935 on 5-HT levels were determined using in vivo microdialysis. Rats which had undergone chronic rTMS showed reduced responses to both challenges, indicating subsensitivity of both the presynaptic 5-HT1A autoreceptors situated somatodendritically in the raphe nuclei and the 5-HT1B autoreceptors situated on nerve terminals. Since such subsensitivity has been demonstrated after other antidepressant treatments, our results indicate that these treatments and rTMS may have a common mechanism of action. PMID- 11006968 TI - Convergent neuroanatomical and behavioural evidence of an amygdala hypothesis of autism. AB - In this study we report a convergence of behavioural and neuroanatomical evidence in support of an amygdala hypothesis of autism. We find that people with high functioning autism (HFA) show neuropsychological profiles characteristic of the effects of amygdala damage, in particular selective impairment in the recognition of facial expressions of fear, perception of eye-gaze direction, and recognition memory for faces. Using quantitative magnetic resonance (MR) image analysis techniques, we find that the same individuals also show abnormalities of medial temporal lobe (MTL) brain structure, notably bilaterally enlarged amygdala volumes. These results combine to suggest that developmental malformation of the amygdala may underlie the social-cognitive impairments characteristic of HFA. This malformation may reflect incomplete neuronal pruning in early development. PMID- 11006969 TI - Ca2+-dependent desensitization of AMPA receptors. AB - The effect of changes in the external concentrations (0.4-10 mM) of Ca2+ ions on AMPA receptors (AMPARs) of different subunit composition was studied on freshly isolated rat brain neurones. Ca2+ produces rapid and reversible voltage independent inhibition of AMPARs. Ca2+-permeable and Ca2+-impermeable AMPARs are equally sensitive to external Ca2+ suggesting that the effect is not addressed to the ion channel. The inhibition of responses evoked by AMPA is significantly larger than those evoked by kainate or glutamate. Cyclothiazide and aniracetam, which are known to prevent AMPAR desensitization, both greatly diminish inhibition of AMPARs by Ca2+. Cyclothiazide is more potent than aniracetam in both preventing of AMPAR desensitization and protecting against the Ca2+ inhibitory effect on hippocampal pyramidal cells. On giant cholinergic interneurones of striatum, aniracetam but not cyclothiazide significantly prevents inhibition by Ca2+. This agrees with available data on relative abundance of flip and flop splice variants in these cell types. The results suggest that Ca2+ may allosterically increase AMPA receptor desensitization independently on subunit composition and splice variants. PMID- 11006971 TI - Two distinct components of initial glutamate release synchronized with anoxic depolarization in rat global brain ischemia. AB - Numerous reports have suggested that anoxic depolarization is a critical event in the pathogenesis of cerebral ischemia. Extracellular glutamate concentration ([Glu]e) is closely related to the pathogenesis of ischemia. Therefore, these pathogenic mechanisms merit study, especially the relationship between [Glu]e elevation and the ionic basis of early changes in membrane potential after ischemic insult in vivo. It is often presumed from electrophysiological studies that a causal relationship exists between impaired glutamate uptake and/or progressive glutamate increase and anoxic depolarization, but few in vivo reports have found any sign of a progressive increase of [Glu]e elevation preceding anoxic depolarization. Recently, we reported the application of an oxygen independent real-time technique for monitoring glutamate levels in the extracellular space during in vivo ischemia, and demonstrated that the massive glutamate release during ischemia is biphasic. In the present study, using this real-time monitoring system, we carried out a more detailed analysis of the initial events in the first phase of glutamate release during ischemia-induced anoxic depolarization. The shape of the rising slope that forms the peak of the first phase suggested two components. The second component was approximately 10 times steeper than the first, with two different components of the rise on the way to the peak of the biphasic [Glu]e elevation. This is the first report to demonstrate these components of the initial glutamate peak, and suggests a progressive second component of the [Glu]e increase preceding Ca2+-dependent release from synaptic vesicles with anoxic depolarization, in vivo. PMID- 11006970 TI - nNOS inhibitors attenuate methamphetamine-induced dopaminergic neurotoxicity but not hyperthermia in mice. AB - Methamphetamine (METH)-induced dopaminergic neurotoxicity is associated with hyperthermia. We investigated the effect of several neuronal nitric oxide synthase (nNOS) inhibitors on METH-induced hyperthermia and striatal dopaminergic neurotoxicity. Administration of METH (5 mg/kg; q. 3 h x 3) to Swiss Webster mice produced marked hyperthermia and 50-60% depletion of striatal dopaminergic markers 72 h after METH administration. Pretreatment with the nNOS inhibitors S methylthiocitrulline (SMTC; 10 mg/kg) or 3-bromo-7-nitroindazole (3-Br-7-NI; 20 mg/kg) before each METH injection did not affect the persistent hyperthermia produced by METH, but afforded protection against the depletion of dopaminergic markers. A low dose (25 mg/kg) of the nNOS inhibitor 7-nitroindazole (7-NI) did not affect METH-induced hyperthermia, but a high dose (50 mg/kg) produced significant hypothermia. These findings indicate that low dose of selective nNOS inhibitors protect against METH-induced neurotoxicity with no effect on body temperature and support the hypothesis that nitric oxide (NO) and peroxynitrite have a major role in METH-induced dopaminergic neurotoxicity. PMID- 11006972 TI - A moderate but not total decrease of mitochondrial membrane potential triggers apoptosis in neuron-like cells. AB - The effects of various degrees of perturbation of the mitochondrial membrane potential (mt delta psi) on apoptosis was investigated by intensified fluorescence digital-imaging microscopy on neuron-like cells, ND7. Mt delta psi was either decreased by 40% by the protonophore carbonyl cyanide p trifluoromethoxyphenylhydrazone (FCCP 100 nM, 15 min) or completely collapsed (FCCP 10 microM, 60 min). A moderate decrease of mt delta psi induced a reduction of mitochondrial NADH, followed by exposure of phosphatidyl serine and then by chromatin condensation, 36% of nuclei being condensed 60 min after FCCP treatment. During these stages, mitochondrion morphology was fully preserved. In contrast, no chromatin condensation was observed after a rapid and total dissipation of mt delta psi. These results suggest that a partial decrease of mt delta psi would allow mitochondrial functions required to trigger apoptosis to be sustained. PMID- 11006973 TI - In vivo imaging of activated microglia using [11C]PK11195 and positron emission tomography in patients after ischemic stroke. AB - Neuroprotective strategies are currently being developed for stroke patients. Although the focus is on the development of early treatment the importance of late pathogenetic events is increasingly recognized. To investigate the microglial reaction in stroke we used a marker for activated microglia, [11C]PK11195, and PET in five patients with ischemic stroke 5-53 days after infarction. In one patient serial measurements were made. We demonstrated in each individual and at each point in time that a microglial reaction takes place in the area where T1 weighted MRI (magnetic resonance imaging) shows intensity changes. We consider this PET method as a promising tool to study the late pathogenetic consequences of cerebral infarction and to evaluate neuroprotective strategies with respect to the consequences of the microglial activation. PMID- 11006974 TI - Ischemic tolerance preserves propagation of membrane depolarization. AB - The functional integrity of the synaptic connections within the hippocampus in gerbils that had acquired ischemic tolerance was investigated. The propagation of membrane depolarization across the hippocampus in response to electrical stimulation of CA1 was monitored with the use of a high speed optical recording technique. In comparison to control slices, propagation was significantly depressed and depolarization was shortened in slices from gerbils subjected to 5 min of ischemia. Hippocampal slices from gerbils who were preconditioned with prior sublethal ischemia demonstrated only a slight reduction in propagation. The duration of depolarization was longer than that of ischemia group. These findings suggest that ischemia induces a functional disturbance of synaptic transmission and membrane depolarization. Ischemic preconditioning significantly reduced the extent of this functional disturbance. PMID- 11006975 TI - Immunohistochemical study of nucleoporin p62 in the hippocampus and hypothalamus of the rat brain. AB - We immunohistochemically studied the distribution of nucleoporin p62 in the hippocampus and hypothalamus of rat brain. Previous reports have shown the presence of p62-immunoreactivity (ir) in the nuclear rim in the non-neuronal cells, but the present study showed that of p62-ir within the nucleus in addition to the nuclear rim in the neuronal cells of the hippocampus and hypothalamic nuclei; in these areas the glucocorticoid receptor (GR) undergoes nucleocytoplasmic translocation determined by ligand. We analyzed the expression of p62-ir after adrenalectomy (ADX). ADX changed the localization of GR-ir from the nucleus to the cytoplasm, but did not change the localization or immunoreactivity of p62, suggesting that nucleoporin p62 is stable regardless of intracellular signal transduction between the cytoplasm and the nucleus. PMID- 11006976 TI - A cortical region sensitive to auditory spectral motion. AB - The functional architecture of human auditory cortex is still poorly understood compared with that of visual cortex, yet anatomical and electrophysiological studies in non-human primates suggest that the auditory cortex also might be functionally specialized, in a model of parallel and hierarchical organization. In particular, spectral changes such as the formant transitions of speech, or spectral motion (SM) by analogy with visual motion, could be processed in specialized cortical regions. In this study, positron emission tomography (PET) was used to identify which auditory cortical region are involved in SM analysis. We found that a bilateral secondary auditory cortical region, located in the caudal-lateral belt of auditory cortex, was more sensitive to auditory stimuli containing spectral changes than to matched stimuli with a stationary spectral profile. This result suggests that analogies between sensory systems could prove useful in the research into the functional organization of the auditory cortex. PMID- 11006977 TI - p53 and Bax implication in NMDA induced-apoptosis in mouse hippocampus. AB - Seven days after in vivo intrahippocampal administration of NMDA, 3'-OH DNA fragmentations and Bax protein expression were detected in hippocampal neurons of p53+/+ but not p53-/- transgenic mice. Interestingly, neurons showing pycnosis, an early apoptotic phenomena, were present in all genotypes. These results confirm that apoptotic 3'OH DNA fragmentations and Bax protein induction during NMDA-induced apoptosis in adult hippocampal neurons are p53 dependent. PMID- 11006978 TI - Activity patterns of human somatosensory cortex adapt dynamically to stimulus properties. AB - Long-term synchronous tactile stimulation of two sites of the body results in integrated, overlapping cortical representations whereas asynchronous stimulation leads to segregated representations. To investigate the cortical capacity to adapt dynamically to stimulation properties 22 subjects were stimulated at digits 1, 3 and 5 of both hands in either random or fixed order. Changes in the functional organization of the somatosensory cortex were inferred by neuromagnetic source analysis based on somatosensory evoked magnetic fields. Compared to the stimulation in random sequence, the stimulation in fixed order revealed a reduction in distance between the cortical representation of D1 and D3. We conclude that the pattern of activation in the somatosensory cortex adapts dynamically to the spatio-temporal characteristics of the stimuli. PMID- 11006979 TI - Fish inner ear otoliths stop calcium incorporation after vestibular nerve transection. AB - Previous investigations revealed that the growth of fish inner ear otoliths (otolith size and calcium incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In a search for the regulating unit, the vestibular nerve was unilaterally transected in neonatal swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation and thus otolith growth ceased on the operated head sides, indicating that the brain is significantly involved in regulating otolith growth. PMID- 11006981 TI - Role of nitric oxide in rat locus coeruleus in hypoxia-induced hyperventilation and hypothermia. AB - The locus coeruleus modulates the ventilatory and thermoregulatory response to hypoxia and contains nitric oxide synthase. Therefore, we examined the effects of L-NAME unilaterally microinjected into the locus coeruleus on hypoxic hyperventilation and hypothermia. Ventilation and body temperature were measured before and after microinjection of L-NAME (100 nmol/0.5 microl) into the locus coeruleus, followed by hypoxia. Control rats received microinjection of D-NAME (an inactive enantiomer of L-NAME). Under normoxia, L-NAME treatment did not affect ventilation or body temperature. D-NAME did not affect hypoxia-induced hyperventilation and hypothermia. L-NAME treatment reduced the ventilatory response to hypoxia but did not affect hypoxia-induced hypothermia. These data suggest that nitric oxide in the locus coeruleus is involved in the ventilatory response to hypoxia, exercising an inhibitory modulation on the locus coeruleus neurons, but plays no role in hypoxia-induced hypothermia. PMID- 11006980 TI - The representation of articulation in the primary sensorimotor cortex. AB - Bilabial, alveolar and velar sounds are produced at different locations around the oral cavity. fMRI was used to localize cortical representation of articulation in seven healthy subjects during repetitive lip and vertical tongue movements as well as repetitive articulation of /pa/ (bilabial), /ta/ (alveolar), /ka/ (velar consonants) and a combination of all three in /pataka/. Centers of gravity (COG) analysis of fMRI activation in the primary motor (M1) and sensory cortex (S1) revealed /pa/- adjacent to lip representation and /ta/- articulation to tongue representation. The articulation of /pataka/ showed an activation pattern with a combination of the two M1/S1-components and additional activation in the supplementary motor area. PMID- 11006982 TI - Remote discharges in the posterior language area during basal temporal stimulation. AB - A 23-year-old woman with refractory complex partial seizures underwent implantation of subdural grid electrodes over the left hemisphere to map epileptic foci and language function. Aphasic symptoms occurred during stimulation of the basal temporal area, which were always associated with intrastimulus remote discharges (ISRDs) in the classical posterior language area. No sequential language deficits occurred after anterior temporal lobectomy including the basal temporal area. These results suggest a close functional relationship between the basal temporal area and posterior language area in patients with temporal lobe epilepsy. ISRDs may explain the paradoxical observation that resection of the basal temporal language area results in no language deficits. PMID- 11006983 TI - Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation. AB - We tested the hypothesis that transplantation of bone marrow stromal cells (MSCs) into the spinal cord after a contusion injury promotes functional outcome. Rats (n = 31) were subjected to a weight driven implant injury. MSCs or phosphate buffered saline was injected into the spinal cord 1 week after injury. Sections of tissue were analyzed by double-labeled immunohistochemistry for MSC identification. Functional outcome measurements using the Basso-Beattie-Bresnehan score were performed weekly to 5 weeks post-injury. The data indicate significant improvement in functional outcome in animals treated with MSC transplantation compared to control animals. Scattered cells derived from MSCs expressed neural protein markers. These data suggest that transplantation of MSCs may have a therapeutic role after spinal cord injury. PMID- 11006984 TI - Neurite outgrowth in developing mouse spinal cord neurons is modulated by glycine receptors. AB - The effect of glycine receptor activation on neurite outgrowth and survival was studied in 5 DIV (days in vitro) spinal neurons. These neurons were depolarized by spontaneous synaptic activity and by glycine, but not by glutamate. These responses were accompanied by increases in intracellular calcium concentration measured with Indo-1 and Fluo-3. Glycine (100 microM, 48 h) increased (46 +/- 6%) the number of primary neurites and total neuritic length. This effect was mediated by synaptic activity and calcium influx because TTX (1 microM) and nimodipine (4 microM) blocked the stimulatory effect of glycine. Neuronal survival, on the other hand, was not affected. This study shows for the first time the modulatory effect of glycine receptors on spinal neuron development. PMID- 11006985 TI - The neural correlates of orienting: an integration of fMRI and skin conductance orienting. AB - In fMRI studies, the averaging of neural activity across multiple trials might obscure important psychophysiological subprocesses. The orienting response (OR) is a distinctive subprocess signalling the active orientation of attention towards potentially significant events. We sought to elucidate fMRI activity associated with visual stimuli that did or did not evoke simultaneously recorded electrodermal ORs (using customised skin conductance recording). 'With-OR' stimuli were associated with significant activity in the hippocampus, anterior cingulate and ventromedial prefrontal cortex. Averaged analysis revealed activity only in the expected visual circuits. Our results suggest that potentially significant stimuli (with-OR) activate different functional networks to familiar (without-OR) stimuli, and that orienting may therefore be an informative subprocess to consider in cognitive fMRI studies. PMID- 11006986 TI - PTX-sensitive and -insensitive synaptic modulation at the frog neuromuscular junction. AB - Pharmacological manipulations were used to examine the role of G proteins in modulating synaptic transmission at the frog neuromuscular junction. Pertussis toxin (PTX, a G protein antagonist) increased end-plate potential (epp) amplitude but had no effect on the amplitude or frequency of miniature end-plate potentials. Mastoparan (a G protein agonist) decreased epp amplitude, while suramin (an antagonist) increased epp amplitude. The results suggest that PTX sensitive G proteins tonically modulate synaptic transmission by reducing the amount of transmitter released in response to presynaptic action potentials. We also showed that endogenous ATP decreased transmitter release via P2 receptor in a PTX-insensitive manner. Thus, at least two distinct mechanisms regulate neuromuscular transmission; one is coupled to PTX-sensitive G proteins and the other is not. PMID- 11006987 TI - Two mirror-image sensory maps with domain organization in the mouse main olfactory bulb. AB - The glomerular sheet in the olfactory bulb (OB) provides an olfactory sensory map identifying which odorant receptors (ORs) in the nose are activated by inhaled odorants. How are the glomeruli spatially arranged in the OB? Using OCAM and neuropilin-1 (NP1) as molecular markers for target glomeruli of distinct subsets of olfactory axons, we demonstrate here that glomeruli are parceled into topographically distinct domains. Spatial arrangement of these domains suggests that each OB contains two mirror-image maps of the glomeruli. In situ hybridization shows that the glomeruli representing the same OR are symmetrically arranged; one in a domain in the lateral hemisphere and the other in a corresponding domain in the medial hemisphere of the OB. These results suggest that OB contains two symmetrical OR maps with similar domain organization. PMID- 11006988 TI - Conduction velocity of the human spinothalamic tract as assessed by laser evoked potentials. AB - To study the conduction velocity of the spinothalamic tract (STT) we delivered CO2 laser pulses, evoking pinprick sensations, to the skin overlying the vertebral spinous processes at different spinal levels from C5 to T10 and recorded evoked potentials (LEPs) in 15 healthy human subjects. These stimuli yielded large-amplitude vertex potentials consisting of a negative wave at a peak latency of about 200 ms followed by a positive wave at a peak latency of about 300 ms. The mean conduction velocity of the STT was 21 m/s, i.e. higher than the reported velocity of the corresponding primary sensory neurons (type II AMH). Because dorsal stimulation readily yields reproducible brain LEPs, we expect this technique to be useful as a diagnostic tool for assessing the level of spinal cord lesions. PMID- 11006989 TI - Age-related decline in the functional response of striatal group I mGlu receptors. AB - In order to verify whether striatal group I metabotropic glutamate (mGlu) receptors undergo functional alteration in ageing, the effects induced by the selective agonist 3,5-dihydroxyphenylglycine (DHPG) in the striatum of young (3 months) and aged (24-25 months old) rats were compared. The ability of DHPG to stimulate phosphoinositide (PI) hydrolysis (striatal slices), to influence striatal dopamine release (in vivo microdialysis) and to potentiate the effects of NMDA on extracellular field potential amplitude (extracellular recordings on striatal slices) was reduced in the striatum of old vs young rats. These results show an age-dependent reduction in the functional response of striatal group I mGlu receptors, which may be one of the factors underlying the reduced ability aged striatum to integrate information. PMID- 11006990 TI - Frontal lobe damage and thalamic volume changes. AB - The aim of this study was to investigate whether frontal lobe damage affects thalamic volume in humans. Ipsilateral and contralateral thalamic areas were measured in 0.5T T1-weighted sagittal magnetic resonance images in 12 patients, first at the time of their surgery for relief of a unilateral frontal lobe brain tumor and at follow-up approximately 2 years later. A 5% decrease in ipsilateral and 4.5% increase in contralateral thalamic area was found over time (F(1,11) = 6.15, p < 0.05). We conclude that unilateral frontal lobe damage results in a decrease in the ipsilateral thalamus and an increase in the contralateral thalamus in humans in vivo. The findings may have implications for the interpretation of the reported changes in thalamic volume in neuropsychiatric diseases. PMID- 11006991 TI - Inhibition of return and oculomotor control in the blind. AB - Our attentional systems orient reflexively to novel environmental stimuli. Such attentive orienting is typically followed by a prolonged period of inhibition, known as inhibition of return (IOR), thought to be linked to the eye movement system. It is widely believed that IOR may provide a tagging mechanism that prevents perseveration, and thus facilitates attentional search. Using a tactile variant of the peripheral spatial cuing paradigm, we show IOR in congenitally blind adults and in an individual who had no eyes. These results demonstrate for the first time that spatial IOR can occur in the absence of oculomotor control. PMID- 11006992 TI - Is there a distortion of body projection in extracorporeal space in neglect patients? AB - Neglect patients often present with a rightward shift of the direction straight ahead of body midline. We investigated whether directions ahead of other body parts were similarly shifted or were differentially affected. Right brain-damaged patients, with and without neglect, and normal subjects had to indicate by manual pointings, the directions straight-ahead of the navel, the nose, the left and right shoulders. In neglect patients, constant errors varied across directions with a decrease in magnitude from left to right, suggesting a compression of body projection in extracorporeal space. Moreover, the projection of the left hemibody was smaller than the right in all subjects, with and without neglect. This unexpected finding is discussed with regard to other spatial asymmetries observed in normal subjects. PMID- 11006993 TI - A case of musical agraphia. AB - Damage to the left upper parietal lobule causes pure agraphia. However, we experienced a patient who exhibited musical agraphia following such a lesion after the agraphia improved. The patient was a 53-year-old female piano teacher. After surgery, she exhibited agraphia and musical agraphia. There was no expressive amusia, receptive amusia, aphasia, agnosia or apraxia. Fifteen months post-surgery, when her agraphia had resolved, her abilities to read, write, and copy music were evaluated. She could read and write single notes and musical signs, but her ability to write a melody was seriously impaired. Furthermore, the salient impairment was in writing rhythm rather than pitch. She could copy music, but only slowly. We consider her a case of pure musical agraphia. PMID- 11006994 TI - What is extinguished in auditory extinction? AB - Extinction is a frequent sequel of brain damage, whereupon patients disregard (extinguish) a contralesional stimulus, and report only the more ipsilesional stimulus, of a pair of stimuli presented simultaneously. We investigated the possibility of a dissociation between the detection and the identification of extinguished phonemes. Fourteen right hemisphere damaged patients with severe auditory extinction were examined using a paradigm that separated the localization of stimuli and the identification of their phonetic content. Patients reported the identity of left-sided phonemes, while extinguishing them at the same time, in the traditional sense of the term. This dissociation suggests that auditory extinction is more about acknowledging the existence of a stimulus in the contralesional hemispace than about the actual processing of the stimulus. PMID- 11006995 TI - The antifungal activity of natamycin toward molds isolated from commercially manufactured poultry feed. AB - The antifungal activity of natamycin, a polyene antifungal compound, was evaluated on molds isolated from commercial poultry feed. The antifungal activity was measured by determination of the minimal inhibitory concentration (MIC) for natamycin on molds growing on semisolid microbiological medium (potato dextrose agar) containing pure natamycin at concentrations ranging from 0 to 200 mg/liter. Natamycin exhibited a high degree of antifungal activity against the 191 isolates of aspergilli used in this study, with average MIC values ranging from 5.08 to 40.1 mg/liter for Aspergillus fumigatus and Aspergillus parasiticus, respectively. Natamycin was also equally effective in inhibiting the growth of nonaflatoxigenic compared with aflatoxigenic isolates of Aspergillus flavus and A. parasiticus. Natamycin was also efficacious against molds other than aspergilli, with MIC values ranging from 2.15 to 5.80 mg/liter for Paecilomyces and Rhizopus spp., respectively. Natamycin exhibited apparent sporicidal activity against spores of toxigenic strains of Fusarium moniliforme and A. parasiticus but not Penicillium rubrum. This sporicidal activity was evident only when spores were exposed to an in vitro concentration of natamycin of 25 mg/liter or higher for a period of time of at least 12 hr. The growth inhibiting activity of natamycin was more pronounced compared with the sporicidal activity. PMID- 11006996 TI - Detection of antibody to turkey coronavirus by antibody-capture enzyme-linked immunosorbent assay utilizing infectious bronchitis virus antigen. AB - An antibody-capture enzyme-linked immunosorbent assay (ELISA) for detection of antibody to turkey coronavirus (TCV) utilizing infectious bronchitis virus (IBV) antigen was developed. Anti-TCV hyperimmune turkey serum and normal turkey serum were used as positive or negative control serum for optimization of the ELISA system. Goat anti-turkey immunoglobulin G (light plus heavy chains) conjugated with horseradish peroxidase was used as detector antibody. The performance of the ELISA system was evaluated with 45 normal turkey sera and 325 turkey sera from the field and the cutoff point was determined. Serum samples of turkeys experimentally infected with TCV collected sequentially from 1 to 63 days postinfection were applied to the established antibody-capture ELISA using IBV antigens. The optimum conditions for differentiation between anti-TCV hyperimmune serum and normal turkey serum were serum dilution at 1:40 and conjugate dilution at 1:1600. Of the 325 sera from the field, 175 were positive for TCV by immunofluorescent antibody (IFA) assay. The sensitivity and specificity of the ELISA relative to IFA test were 93.1% and 96.7%, respectively, based on the results of serum samples from the field turkey flocks using the optimum cutoff point of 0.18 as determined by the logistic regression method. The ELISA values of all 45 normal turkey sera were completely separated from that of IFA-positive sera. The ELISA results of serum samples collected from turkeys experimentally infected with TCV were comparable to that of the IFA assay. Reactivity of anti rotavirus, anti-reovirus, anti-adenovirus, or anti-enterovirus antibodies with the IBV antigens coated in the commercially available ELISA plates coated with IBV antigens could be utilized for detection of antibodies to TCV in antibody capture ELISA. PMID- 11006997 TI - Morphologic observations on respiratory tracts of chickens after hatchery infectious bronchitis vaccination and formaldehyde fumigation. AB - The histologic changes in the respiratory tracts of chickens were evaluated after hatchery fumigation with 40% formaldehyde vapors and vaccination against infectious bronchitis virus with live attenuated vaccine (Massachusetts serotype). One-day-old chickens were housed in four isolation units in controlled environmental conditions, fed and watered ad libitum, and separated into four groups: 1) fumigated and vaccinated birds (FV group); 2) nonfumigated and vaccinated birds (NFV group); 3) fumigated and nonvaccinated birds (FNV group); and 4) control group (C group). All birds were tested to be free from Mycoplasma gallisepticum and Mycoplasma synoviae. After necropsy on the first, eighth, and twenty-sixth days after birth, samples from tracheal upper portion and lungs were conventionally processed for light, scanning, and transmission electron microscopy. Tissue response was monitored by microscopic examination of trachea and lung. On the first day of observation, fumigated and vaccinated birds (FV group) showed extensively damaged tracheal epithelium with exfoliated areas and some active glands with electrodense granules, and in the lung, the primary bronchi epithelium had disorganized cilia and abundant lymphocytes, with emphysematous areas in tertiary bronchus. On day 8 after vaccination, cubical and cylindrical tracheal cell proliferation was observed, and on day 26, ciliated columnar epithelium was almost regenerated with heterophil corion infiltration, and hyaline cartilage nodules appeared in parabronchi. The nonfumigated and vaccinated birds (NFV) revealed less injury on the epithelial surface and a more rapid response to epithelial regeneration than the in only fumigated animals (FNV). The control group did not show remarkable morphologic changes. Postvaccinal and fumigation effects on the upper respiratory tract were temporary, whereas in lungs, increased emphysema, cartilage nodules in the interchange zone, and general lymphocyte infiltration had caused intensive injury. PMID- 11006998 TI - A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin. AB - In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples. PMID- 11006999 TI - Sequence analysis of the hemagglutinin gene of H9N2 Korean avian influenza viruses and assessment of the pathogenic potential of isolate MS96. AB - Sequence analysis of the hemagglutinin (HA) gene of five Korean H9N2 avian influenza virus (AIV) isolates showed that these viruses were closely related and possibly came from the same source. Phylogenetic analysis of the HA1 subunit of H9 subtype isolates revealed that Korean AIV isolates were different from isolates from the poultry markets in Hong Kong in 1997. None of the Korean AIVs had multiple basic amino acids at the HA cleavage site that confer high pathogenicity to some H5 and H7 AIVs. Phylogenetic analysis of the nucleoprotein and matrix gene demonstrated that Korean isolates cluster with Eurasian origin AIVs. The pathogenic potential of one of the isolates (MS96) was assessed after several passages in 14-day-old embryonated chicken eggs (ECE). Fourteen-day-old ECE derivatives of MS96 showed increased HA titer and embryo mortality in eggs; this was apparent after the third passage in 14-day-old ECE. Sequence analysis of the cleavage site of MS96 after the third and tenth passages in 14-day-old ECE revealed no changes in the amino acid sequence. The pathogenicity of MS96 after the tenth passage in 14-day-old eggs (MS96p10(ECE14)) was tested with 4-wk-old specific-pathogen-free chickens. The 14-day-old derivative, MS96p10(ECE14), showed wider tissue tropism and induced more severe clinical signs than the parent virus. Furthermore, after intranasal inoculation of 86-wk-old broiler breeders and 30-wk-old layers, the MS96p10(ECE14) derivative induced more severe signs of depression than the parent virus as well as a transient drop in egg production. PMID- 11007000 TI - Protective immunity to infectious bronchitis in broilers vaccinated against Marek's disease either in ovo or at hatch and against infectious bronchitis at hatch. AB - Two experiments were conducted using commercial broiler chickens to determine if Marek's disease (MD) vaccines HVT/SB-1 and HVT plus CVI-988 given either in ovo or at hatch adversely affected the efficacy of infectious bronchitis (IB) vaccines (Ark and Mass serotypes) given by eyedrop on the day of hatch. Nonvaccinated negative controls and controls that received only IB vaccines were included in each study. Birds were challenged with either infectious bronchitis virus (IBV) Mass-41 or IBV Ark-99 on either day 26 or 27 of age. Protection was assessed 5 days post-IBV challenged by virus isolation from the trachea. The day of hatch mean antibody titer to IBV was 12,668 +/- 4704 and 2503 +/- 3243 by enzyme-linked immunosorbent assay in experiments 1 and 2, respectively. In each study, nonvaccinated controls had a significantly higher (P < or = 0.05) incidence (88%-100%) of IBV challenge virus isolation than did controls vaccinated for IB but not for MD. Analysis of data from both studies showed that protection to IB in groups that received only IB vaccines at hatch ranged from 55.0% to 77.3%, whereas protection to IB in groups receiving both MD and IB vaccines ranged from 50.0% to 95.5%. In both experiments and within IBV challenge serotype, broilers given MD vaccines (in ovo or at hatch) and IB vaccines at hatch had protection rates to IBV challenges that were not significantly less (P < or = 0.05) than IB protection rates of groups that received only IB vaccines at hatch. Analysis of these data shows that administration of high-titered MD vaccines either in ovo or at hatch did not affect the efficacy of an IB vaccination (serotypes Ark and Mass) given by eyedrop at hatch. PMID- 11007001 TI - Importance of Escherichia coli infection in ascites in broiler chickens shown by experimental production. AB - Common commercial strain male broilers aged 14 days were intratracheally inoculated with 0.2 ml of 1.2 x 10(6) colony-forming units of Escherichia coli in nutrient broth and kept in a cool environment during the experiment. Ascites was produced in five surviving and two dead birds out of 50 but not in 50 mock infected control birds. Among the 40 survivors that were infected, the erythrocyte packed cell volume (PCV) of the 10 birds with pericarditis was the same as in 21 grossly normal birds, although that of the four birds with enlarged right ventricle (RV) was high. The pericarditis caused by E. coli septicemia was not the primary cause of ascites. However, the PCV was high in some of the survivors with an enlarged RV without pericarditis, indicating overload due to the lung lesion. These data suggested that some of the birds with an enlarged RV, caused by supplying blood that was insufficiently oxygenated for the body size, suffered from ascites. PMID- 11007002 TI - Ornithobacterium rhinotracheale infection in turkeys: immunoprophylaxis studies. AB - Ornithobacterium rhinotracheale has been shown to cause serious clinical illness and is a significant concern to the turkey industry because of its potential economic impact. In this study, 6-wk-old turkeys were vaccinated intranasally with a live or subcutaneously with a killed O. rhinotracheale vaccine. At 14 or 21 wk of age, the birds were challenged intratracheally with live O. rhinotracheale. Airsacculitis and pneumonia occurred less frequently in vaccinated birds than in unvaccinated birds after challenge with O. rhinotracheale. Ornithobacterium rhinotracheale was recovered from unvaccinated, challenged birds but not from vaccinated, challenged or from unchallenged birds. Thus, turkeys inoculated with live or killed O. rhinotracheale vaccine were protected from pathologic changes. PMID- 11007003 TI - A comparative study of the pathogenesis of malabsorption syndrome in broilers. AB - Five malabsorption syndrome (MAS) homogenates from The Netherlands and Germany were used to reproduce MAS in broilers. We studied the histopathology after inoculation of 1-day-old broiler chicks and the agents that might be involved. Generally, the MAS homogenates induced signs that differed in severity and pathobiology. We could distinguish and classify the inoculated groups best by histopathology: proventriculitis, lesions in the small intestines in combination with proventriculitis, or lesions of the small intestines only. Lesions in the small intestine had more impact on weight gain depression than lesions in the proventriculus. In three out of five inoculated groups, microscopic lesions of the pancreas were found. Reovirus was detected in the inoculated groups by virus isolation and seroconversion, and reoviral antigen was detected by immunohistochemistry of the small intestine. Also, enteroviruslike particles were detected in three of the five inoculated groups, although not in the most affected group. Additionally, bacteriophages and bacteria (hemolytic Escherichia coli, Pasteurella hemolytica, and Enterococcus durans) were isolated from inoculated chicks. The role these agents play in pathogenesis of MAS is still unsolved. PMID- 11007004 TI - Emergence of subtype strains of the Arkansas serotype of infectious bronchitis virus in Delmarva broiler chickens. AB - Infectious bronchitis virus (IBV) field isolates of the Arkansas (Ark) serotype were identified by reverse transcription-polymerase chain reaction (RT-PCR) as the most common serotype isolated from 1993 to 1997. These isolates were recovered from broiler flocks with respiratory disease raised on the Delmarva peninsula in spite of Ark vaccination in the region. For the purposes of investigating this apparently paradoxical finding, five RT-PCR Ark-positive field isolates recovered in 1995 and 1996 were selected for further characterization. The isolates were compared with Ark reference strains by reciprocal virus neutralization (VN) in embryonated eggs, S-1 gene sequence analysis, and challenge of immunity studies in specific-pathogen-free (SPF) chickens. Antigenic (VN) comparisons and S-1 gene analysis confirmed that the five RT-PCR Ark positive field isolates were of the Ark serotype but also revealed that the viruses could be readily distinguished from Ark reference strains. Four of the isolates (Ark/213/96, Ark/15C/96, Ark/1529/95, Ark/1534/95) were found to have higher antigenic relatedness percentages to each other (95%-100%) than to Ark reference strains DPI (52%-72%) and Georgia variant (Georgia var) (53%-68%) by VN. Another isolate, Ark/1535/95, was found to differ antigenically from the other four RT-PCR Ark-positive field isolates (34%-61%), Ark DPI (44%), and Georgia var (43%) strains. The trends in the S-1 gene sequencing results were similar to those observed for the VN findings. Isolates Ark/213/96, Ark/15C/96, Ark/1529/95, and Ark/1534/95 demonstrated a higher degree of predicted S-1 amino acid similarity to each other (96.5%-98.7%) than to Ark DPI (92.4%-93.7%), Ark 99 (93.2%-94.7%), and Georgia var (89.3%-90.8%). Ark/1535/95 S-1 amino acid similarity values were lower compared with those of the other four RT-PCR Ark positive field isolates (93.4%-94.8%), Ark DPI (91.9%), Ark 99 (93.0%), and Georgia var (88.7%). Furthermore, the isolates could be distinguished from the Ark reference strains by a characteristic sequence polymorphism, a six-nucleotide deletion encoding amino acids 57 (Asp) and 58 (Asp) in hypervariable region 1 of S-1. On the basis of the VN and sequencing findings, isolates Ark/213/96, Ark/15C/96, Ark/1529/95, and Ark/1534/95 were considered to be a single subtype of the Ark serotype. The fifth isolate, Ark/1535/95, may constitute another subtype of the Ark serotype. Vaccination of SPF chickens with a high-titering commercially available live vaccine containing the Ark DPI strain provided solid protection (>90%) against challenge with the RT-PCR Ark-positive field isolates. Immunization of SPF chickens with Ark/213/96 produced 100% protection against challenge with the homologous strain, as well as isolates Ark/1535/95 and Ark 99 but lower levels of protection against Ark DPI (58%) and Georgia var (55%). Primers for RT-PCR were designed to distinguish between the Ark subtypes and the Ark reference strains on the basis of the characteristic six-nucleotide deletion identified in the S-1 gene of the Ark subtypes. Retrospective analysis of RT-PCR Ark-positive isolates found that the Ark subtypes existed as early as 1992 in Delmarva broilers and became prevalent by 1995. With RT-PCR, restriction fragment length polymorphism analysis, and DNA sequencing techniques, the presence of Ark subtype viruses was demonstrated in two commercial Ark DPI strain vaccines and in our Ark DPI laboratory stocks that were the original source of the virus used for vaccine development. The demonstration of the Ark subtype and reference strains in the Ark DPI strain is evidence of the existence of IBV quasispecies. Factors possibly influencing the emergence of the Ark subtype in commercial broilers are discussed. PMID- 11007005 TI - Characterization of infectious bronchitis viruses isolated from outbreaks of disease in commercial flocks in Brazil. AB - Fifteen isolations of infectious bronchitis (IB) virus were made from a total of 126 Brazilian poultry flocks of all ages that were examined. These flocks (14 chicken and 1 quail) were experiencing a variety of IB-like conditions including respiratory disease, digestive and kidney problems, and drops in egg production. One of the isolates was of the Massachusetts serotype. The remainder were examined by means of cross-neutralization tests in tracheal organ cultures and were shown to belong to at least four antigenic groups, all different from ones described previously in other countries. Some, but not all, of the flocks from which they were isolated had been vaccinated against IB with vaccines of the Massachusetts serotype. In vivo protection studies showed that the MA5 vaccine (of the Massachusetts serotype) protected well against challenge with four of these isolates, representing the different serotypes reported in this study. PMID- 11007006 TI - Pathogenicity of a low-virulence duck virus enteritis isolate with apparent immunosuppressive ability. AB - Duck enteritis virus (DEV) was isolated from commercial 2-to-6-wk-old white Pekin ducks experiencing 25%-30% mortality and high morbidity. Secondary infections with Pasteurella multocida, Riemerella anatipestifer, and Escherichia coli were frequently seen in affected ducks. The isolated virus was identical to the prototype DEV by virus neutralization test but differed from the classic DEV by causing lymphoid organ atrophy and inconsistent hemorrhagic lesions in the intestinal annular bands. Attempts to reproduce the disease in white Pekin ducks were unsuccessful until the virulence of the virus was increased by three passages in Muscovy ducklings. Significant thymic atrophy (P < or = 0.001) was detected during the first 10 days postinfection (DPI), but thymus size returned to normal by 17-24 DPI. However, bursal atrophy increased significantly (P < or = 0.001) from 4 DPI until the end of the experiment (39 DPI). Reduction in body weight was significant (P < or = 0.05) between 4 and 6 DPI. There was massive depletion of thymic and bursal lymphocytes with lymphoid necrosis in the thymus, bursa, spleen, and Harderian gland. Eosinophilic intranuclear inclusions were observed in thymus, bursa, spleen, esophagus, cloaca, liver, conjunctiva, and Harderian gland. Occasional intracytoplasmic inclusions were also found scattered in the epithelial cells of conjunctiva, esophagus, bursa of Fabricius, and cloaca. Virus was recovered from experimentally infected ducks from thymus, bursa, spleen, liver, kidneys, trigeminal ganglion, and cloaca during the first 10 days of infection. These findings suggest that a low-virulent DEV can cause a massive lymphoid atrophy and can sustain immunosuppression as noted by the secondary bacterial infection. PMID- 11007007 TI - Characterization of a small round virus associated with the poult enteritis and mortality syndrome. AB - A small round virus (SRV) identified and isolated in our laboratory from intestinal samples of poults affected with the poult enteritis and mortality syndrome was further characterized. The SRV was propagated in turkey embryos and purified by differential and isopycnic ultracentrifugation. The size of the SRV was 30-32 nm in diameter. The buoyant density of the SRV in cesium chloride was between 1.34 and 1.36 g/cm3. It was resistant to chloroform treatment, stable at pH 3.0, and resistant to heat treatment. Attempts to propagate the SRV in turkey embryo kidney, turkey kidney, Caco-2, Vero, and BGM-70 cells were unsuccessful. Analysis of the SRV capsid proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed three polypeptides with molecular weights of 34.5, 31, and 28 kD. Genome analysis of the SRV showed that the SRV had a single-strand RNA genome about 7500 nucleotides in length. Reverse transcription-polymerase chain reactions (RT-PCRs) with primers specific to conserved sequences of enteroviruses yielded products with expected sizes. However, sequence analysis of the RT-PCR products showed that there was no similarity between the sequences and that of enteroviruses. RT-PCR with primers specific to the 3' end of a SRV RNA genome yielded products with expected sizes. These products were sequenced and found to contain 669 nucleotides, excluding the polyadenylated tail. Sequence analysis indicated that the SRV shared 38.18% amino acid identity in the C-terminal capsid precursor protein and 41.26% nucleotide identity of the 3' end of turkey astrovirus RNA genome (Genbank accession no. Y15936). We concluded that the SRV is a member of the astrovirus family. PMID- 11007008 TI - Significance of infectious bursal disease serology in an integrated quality control program under European epidemiologic conditions. AB - In this study performed between 1993 and 1997, infectious bursal disease virus (IBDV) antibody titers and performance data were recorded in a vertically integrated monitoring scheme in order to make a follow-up from day-old parents down to the broilers at slaughter. All measured data were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. It appeared that high and/or uniform antibody titers in the parents were correlated with increased daily weight gain and decreased mortality and slaughterhouse condemnation in the broilers. Antibody titers and their CVs were negatively correlated in broiler parents and their offspring at day-old and even at slaughter. Results indicate that high and uniform antibody titers against IBDV in broiler parents are important for good performance of the broiler offspring, at least under the epidemiologic conditions of this study, which included the presence of very virulent IBDV strains in the field and the sole use of live intermediate vaccines in broilers as well as broiler parents. PMID- 11007009 TI - The effects of ts-11 strain Mycoplasma gallisepticum vaccination in commercial layers on egg production and selected egg quality parameters. AB - Live Mycoplasma gallisepticum (MG) vaccines have been USDA approved and licensed for use in commercial layer chickens since 1988; however, egg production and egg quality data exist only for the F strain of MG. Information pertinent to the effects of ts-11 MG on egg and eggshell quality parameters, as well as egg size distribution, is lacking. In this study, pullets were inoculated at 10 wk of age with ts-11 strain MG and placed in biological isolation units at 10 birds/unit. Hen-day egg production, eggshell strength, Haugh unit score, pimpling incidence, and blood/meat spot incidence were monitored and recorded in each trial through a 45-wk production cycle. Further, eggs from all treatments were collected daily, Monday-Thursday, and individually weighed. Results of this study indicate that no significant difference was observed between the treatments for the parameters measured or for egg size distribution. Therefore, these data should lessen producers' concerns pertaining to the impact of ts-11 strain MG on egg production, egg and eggshell quality parameters, and egg size distribution. PMID- 11007010 TI - Comparison of virus isolation, immunohistochemistry, and reverse transcriptase polymerase chain reaction procedures for detection of turkey coronavirus. AB - A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure and two monoclonal antibody (MAb)-based immunohistochemical procedures were developed for detection of turkey coronavirus (TCV) in tissues and intestinal contents/dropping samples. The RT-PCR, MAb-based fluorescent antibody (FA), and MAb-based immunoperoxidase (IP) procedures were compared with virus isolation (VI) for detection of TCV in experimentally infected turkeys. TCV was detected in experimentally infected turkeys as early as day 1 postexposure (PE) by each of the four detection procedures. TCV was detected as late as day 35 PE by FA or IP and days 42 and 49 PE by VI and RT-PCR, respectively. With VI as a reference, sensitivity and specificity of RT-PCR were 93% and 92%, respectively; specificity of both FA and IP was 96%, and sensitivities were 69% and 61%, respectively. Each of the examined procedures was highly specific, but the RT-PCR procedure was also highly sensitive. These findings demonstrate the utility of both immunohistochemistry and RT-PCR for detection of TCV. In addition, the findings indicate that RT-PCR is a highly sensitive and specific alternative to conventional diagnostic procedures. PMID- 11007011 TI - The commercial production of specific-pathogen-free eggs and chickens: the evolution of an industry. PMID- 11007012 TI - The history of avian reovirus. PMID- 11007014 TI - Redesign of primer and application of the reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism test to the DE072 strain of infectious bronchitis virus. AB - Diagnosis of the DE072 strain of infectious bronchitis virus (IBV) by the reverse transcriptase-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) serotype identification test was not possible because the primer used in the RT-PCR did not amplify the S1 gene of the DE072 strain. The 3' end of the polymerase gene and the 5' end of the S2 gene of the DE072 strain were sequenced and compared with the forward and reverse RT-PCR primers, respectively. A 2-bp mismatch at the 3' end of the reverse primer was found. On the basis of these data, a degenerate primer that could amplify the S1 gene of the DE072 strain as well as eight other serotypes of the virus was synthesized. In addition, we were able to differentiate the DE072 strain from all of the other IBV strains examined by RFLP analysis of the RT-PCR product. PMID- 11007013 TI - Pathogenicity of West Nile virus in chickens. AB - In the fall of 1999, West Nile virus (WNV) was isolated for the first time in the Western Hemisphere during an outbreak of neurologic disease in humans, horses, and wild and zoo birds in the northeastern United States. Chickens are a potential reservoir for WNV, and little is known about the pathogenicity of WNV in domestic chickens. Seven-week-old chickens derived from a specific-pathogen free flock were inoculated subcutaneously with 1.8 x 10(3) 50% tissue culture infectious dose of a crow isolate of WNV in order to observe clinical signs and evaluate the viremic phase, gross and microscopic lesions, contact transmission, and immunologic response. There were no observable clinical signs in the WNV inoculated chickens during the 21-day observation period. However, histopathologic examination of tissues revealed myocardial necrosis, nephritis, and pneumonitis at 5 and 10 days postinoculation (DPI); moderate to severe nonsuppurative encephalitis also was observed in brain tissue from one of four inoculated birds examined at 21 DPI. WNV was recovered from blood plasma for up to 8 DPI. Virus titers as high as 10(5)/ml in plasma were observed at 4 DPI. Fecal shedding of virus was detected in cloacal swabs on 4 and 5 DPI only. The WNV also was isolated from myocardium, spleen, kidney, lung, and intestine collected from chickens euthanatized at 3, 5, and 10 DPI. No virus was isolated from inoculated chickens after 10 DPI. Antibodies specific to WNV were detected in inoculated chickens as early as 5 DPI by the plaque reduction neutralization test and 7 DPI by the indirect fluorescent antibody test. Chickens placed in contact with inoculated chickens at 1 DPI lacked WNV-specific antibodies, and no WNV was isolated from their blood plasma or cloacal swabs throughout the 21 days of the experiment. PMID- 11007015 TI - Newcastle disease and avian influenza A virus in wild waterfowl in South Africa. AB - In an intensive ostrich farming area in South Africa with a history of ostrich influenza outbreaks, we conducted a survey of avian influenza virus (AIV) and Newcastle disease virus (NDV) in wild aquatic birds. During late autumn and winter 1998, the time of year when outbreaks in ostriches typically start to occur, 262 aquatic birds comprising 14 species were sampled and tested for both virus infections. From eight samples, AIV, serotype H10N9, could be isolated. All isolates were apathogenic as determined by the intravenous pathogenicity index (0.00). Conversely, none of 33 sera of these wild birds showed antibodies against H10. However, one bird was found serologically positive for H6 AIV. This AIV serotype was later isolated from ostriches during an avian influenza outbreak in this area. No NDV was isolated although 34 of 46 serum samples contained NDV specific antibodies. This is the first H10N9 isolate to be reported from Africa. In addition, our data support the notion that wild aquatic birds may function as a reservoir for AIV and NDV in South Africa. PMID- 11007016 TI - Humoral immune responses to chicken infectious anemia virus in three strains of chickens in a closed flock. AB - This is a comparative study on seroconversion to chicken infectious anemia virus (CIAV) in a closed flock of specific-pathogen-free chickens undergoing a natural outbreak and after vaccination of some of these flocks with a commercial, live vaccine. The N2a strain (B21B21 haplotype) had the highest seroconversion after natural infection (94%) or vaccination (100%), followed by the P2a strain (B19B19) at 75%-82% seroconversion after natural infection and 85% seroconversion after vaccination. The S13 (B13B13) chickens were 26% seropositive after natural infection and 75% seropositive after vaccination. N2a chickens with polymerase chain reaction (PCR)-positive tissues were 97% seropositive compared to 80%-83% PCR-positive and seropositive for the P2a chickens and only 8% seropositive and PCR-positive for the S13 chickens. Seroconversion occurred at or near sexual maturity after natural infection in seven flocks studied. PMID- 11007018 TI - Bumblefoot and lack of exercise among wild and captive-bred falcons tested in the United Arab Emirates. AB - In the 1990s, bumblefoot posed a major health problem to the falcons in the United Arab Emirates. This retrospective study based on statistical field research showed that in captivity wild falcons need a training frequency of twice a day to reduce the bumblefoot morbidity rate. In this context, they responded very well to free flight in aviaries during the molting season. In contrast, captive-bred falcons did not show a highly significant difference regarding training frequencies. Wild falcons tested fed with pigeons, bustards, and ducks suffered significantly less from bumblefoot compared with those fed a diet of quail. Apart from the disease-reducing impact of beef and mice, captive-bred falcons tested did not show any significant difference regarding the influence of diet on the bumblefoot occurrence. PMID- 11007017 TI - Drag swab efficiency factors when sampling chicken manure. AB - This study examines drag swabbing distance, media for moistening the drag swabs, and site selection when sampling a laying facility by drag swabbing manure piles. Manure piles at a laying facility in California's San Joaquin Valley were sampled with drag swabs over various distances. Samples were cultured for Salmonella spp. with standard laboratory methods, and most probable number calculations. Salmonella spp. counts were expected to be highly variable because of reported clustering. Therefore, total bacteria and Escherichia coli, which were assumed to have a more uniform distribution on the surface of the manure, were additionally used as proxies for Salmonella. Media for moistening the swabs were compared by seeding postswabbing samples with Salmonella typhimurium, and culturing at different delay times. Total bacterial counts were compared between samples that were obtained from either wet or dry surfaces. Numbers of Salmonella spp. and total bacteria peaked within 120 feet of swabbing distance. Higher total bacteria counts were obtained by swabbing wet areas rather than dry areas, but the distance that could be swabbed effectively was shorter in wet areas. Moistening media selected for the swab resulted in statistically different culture counts, but did not show any important difference in maintaining Salmonella viability over a 48-hr period when the samples were kept at refrigerated temperatures. Once swabs became fully loaded with fecal material, bacterial numbers failed to increase with further use. Overuse of a swab may result in failure to detect Salmonella enteritidis on chicken manure if the distribution of this organism is clustered. PMID- 11007019 TI - The sensitivity and specificity of a reverse transcription-polymerase chain reaction assay for the avian pneumovirus (Colorado strain). AB - A reverse transcription-polymerase chain reaction (RT-PCR) assay for the detection of avian pneumovirus (APV), Colorado strain (US/CO), was evaluated for sensitivity and specificity. The single-tube RT-PCR assay utilized primers developed from the matrix (M) gene sequence of the US/CO APV. The RT-PCR amplified the US/CO APV but did not amplify other pneumoviruses, including the avian pneumoviruses subgroups A and B. The RT-PCR was capable of detecting between 10(0.25) mean tissue culture infective dose (TCID50) and 10(-0.44) TCID50 of the US/CO APV. These results have demonstrated that the single-tube RT-PCR assay is a specific and sensitive assay for the detection of US/CO APV. PMID- 11007020 TI - Characterization of a lentogenic Newcastle disease virus isolated from broiler chickens in Japan. AB - Newcastle disease virus (NDV), named MET95, was isolated from a non-vaccinated broiler flock in Japan in 1995. The MET95 strain was determined to be a lentogenic NDV. The strain has the properties of eluting rapidly at 4 C and has low thermostability in hemagglutinating activity with chicken erythrocytes. In these studies, no difference could be found between the MET95 strain and the Hitcher B1 vaccine strain. However, the chickens inoculated with the MET95 strain, as well as chickens that they were in contact with, had a much higher hemagglutination-inhibition antibody response than those inoculated with the B1 strain. Accordingly, the MET95 strain is thought to be a promising candidate as a live ND vaccine strain. In Japan, this is the first report on the isolation of lentogenic NDV from chickens since the paper on the Ishii strain isolated in 1966. PMID- 11007021 TI - Sequence of precursor polyprotein gene (segment A) of infectious bursal disease viruses isolated in Korea. AB - The coding regions of segment A of two recent Korean very virulent (vv) infectious bursal disease virus (IBDV) isolates (KK1 and KSH) and one atypical IBDV isolate (K310) were amplified by reverse transcriptase-polymerase chain reaction, sequenced, and compared with published sequences for IBDV. The overall amino acid sequence similarity of the KK1 and KSH strains compared with foreign vvIBDV strains was between 97.43% and 98.02%. The KK1 and KSH strains, like vvIBDV strains, share unique amino acid residues at positions 222(A), 256(I), 294(I), and 299(S). The sequence of K310 strain was markedly different from other IBDV strains. The K310 strain had 12, 2, and 1 unique amino acid substitutions in the VP2 hypervariable region, VP4, and VP3 gene, respectively, and 3 of 12 substitutions in a VP2 hypervariable region were found in two hydrophilic regions known to be involved in antigenic determination. Also, the K310 strain had 222(S) and 254(S), which were found in variant IBDV strains. The SWSASGS heptapeptide is conserved in all Korean IBDV isolates. By phylogenetic analysis, KK1 and KSH were categorized in one group with foreign vvIBDV isolates, but K310 isolate was categorized in a separate group that was differentiated from the other IBDV strains compared. The K310 strain seemed to be evolved from a separate lineage of IBDV strain. PMID- 11007022 TI - Preparation of a heterogeneous conjugate to detect Mycoplasma iowae by immunofluorescence. AB - Fluorescent antibody technique has been used as a diagnostic tool for the identification of pathogenic avian Mycoplasma species. Fluorescein-conjugated antisera for Mycoplasma iowae prepared from rabbit hyperimmune serum against one serotype (I) has not always been reliable, giving false-negative results. A fluorescein-conjugated antisera was prepared with antibody against the six identified serotypes (I, J, K, N, Q, and R) that comprise the species M. iowae. This heterogeneous conjugate was used to positively identify M. iowae in field isolates and lab strains, while maintaining specificity. Polymerase chain reaction specific for M. iowae was used to determine specificity. PMID- 11007023 TI - The effect of vitamin E on cellulitis in broiler chickens experiencing scratches in a challenge model. AB - Two experiments are described; each experiment contained five treatments with each treatment consisting of a specific diet and vitamin E at 8.82 mg, 41.89 mg, 74.96 mg, 108.03 mg, or 141.10 mg vitamin E per kilogram of feed. Birds were raised with continuous feed containing the various levels of vitamin E available throughout the experiment. At 4 wk of age, the birds were scratched on the breast and placed onto avian cellulitis Escherichia coli-seeded litter. One week later, the birds were euthanatized and lesion presence was noted. There appeared to be a positive correlation between vitamin E and the inhibition of cellulitis formation when the birds were fed a diet containing 74.96 mg vitamin E/kg feed. Conflicting results were seen in the two experiments when the birds were fed 41.89 and 108.03 mg vitamin E/kg feed. Both experiments had a high incidence of cellulitis in birds whose diets consisted of 141.10 mg vitamin E/kg feed. PMID- 11007024 TI - Deposition of phage type 4 and 13a Salmonella enteritidis strains in the yolk and albumen of eggs laid by experimentally infected hens. AB - Because egg yolk and albumen differ substantially in their abilities to support bacterial growth, the initial level and location of Salmonella enteritidis deposition are critical for determining whether proposed standards for refrigerating eggs are likely to protect public health by preventing extensive microbial multiplication. In the present study, three groups of laying hens were infected with oral doses of approximately 10(9) cells of different S. enteritidis strains (two were phage type 4 and one was phage type 13a) in two replicate trials. For all three S. enteritidis strains, the incidence of yolk contamination (approximately 2.5% overall) was significantly greater than the incidence of albumen contamination (approximately 0.5% overall). The phage type 13a strain was less often isolated from fecal samples at 2 wk post-inoculation than were the phage type 4 strains, but no significant differences between strains were observed in the incidence of egg contamination. Most freshly laid contaminated eggs contained fewer than 1 S. enteritidis cell/ml of egg yolk or albumen, and no sample contained more than 67 S. enteritidis cells/ml. PMID- 11007025 TI - Eimeria brunetti and Eimeria necatrix in chickens of Argentina and confirmation of seven species of Eimeria. AB - Ten poultry farms (broiler breeder pullets, layer pullets, and broilers) in the provinces of Entre Rios and Buenos Aires in Argentina were examined for presence of Eimeria spp. Litter samples obtained from flocks 7-11 wk old were taken to the laboratory for oocyst counting and sporulation, then concentrated for inoculation into coccidia-free chickens. Species were identified by prepatent period, oocyst size, location and appearance of lesions in the intestine, microscopic examination of mucosal smears, and histology (to confirm Eimeria brunetti). On this basis, Eimeria praecox was found in two samples, Eimeria mitis in two, Eimeria acervulina in nine, Eimeria maxima in seven, Eimeria necatrix in three, Eimeria tenella in seven, and E. brunetti in four. These results confirm the presence of all seven recognized species of Eimeria in chickens in the Republic of Argentina. PMID- 11007026 TI - Determination of the incidence of Salmonella spp., Campylobacter jejuni, and Clostridium perfringens in wild birds near broiler chicken houses by sampling intestinal droppings. AB - Several methods were evaluated for collecting fecal and intestinal samples from wild birds found near broiler chicken houses. A few intestinal samples and cloacal swabs were obtained from European starlings and house sparrows. Most of the samples collected consisted of wild bird droppings found on or near the houses. Samples were collected from each of four farms of a broiler integrator during a grow-out cycle: a cycle in the summer for farm A, fall for farm B, and spring, summer, fall, and winter for farms C and D. Of the 25 wild bird intestinal and fecal samples collected from a broiler house on farm A during a grow-out cycle in July-August 1997, 24% were positive for Salmonella spp., 4% for Campylobacter jejuni, and 28% for Clostridium perfringens. Of the nine fecal samples collected from broiler house B in a grow-out cycle in September-November 1997, 33% were positive for Salmonella spp., 11% for C. jejuni, and 22% for C. perfringens. For farms C and D, of the 23 samples collected in March-April 1998, 0 were positive for Salmonella spp., 11% for C. jejuni, and 52% for C. perfringens; of 27 samples collected in June-July 1998, 4% were positive for Salmonella spp., 0 for C. jejuni, and 13% for C. perfringens; of 24 samples collected in August-October 1998, 14% were positive for Salmonella spp., 5% for C. jejuni, and 4% for C. perfringens; of 14 samples collected December 1998 January 1999, 0 were positive for Salmonella, 50% for C. jejuni, and 14% for C. perfringens. The incidence of these bacterial enteropathogens in wild birds near the broiler chicken houses suggests that wild birds that gain entry to poultry grow-out houses have the potential to transmit these pathogens to poultry. PMID- 11007027 TI - An outbreak of erysipelas in 2-day-old poults. AB - Systemic erysipelas infection was seen in 2-to-4-day-old poults from three separate ranches owned by the same company. The affected poults were all from the same breeder source; poults from other breeder sources were seemingly unaffected. Mortality increased on days 2 and 3, ranging from 2% to 8.5%. Birds submitted were dehydrated and very weak, with one half of the poults submitted having died during transport to the lab. Gross lesions included swollen, congested livers and spleens, as well as hemorrhagic breast muscle in one case. Toes were swollen and reddish-purple in color. The poults had been toe-trimmed during hatchery processing using a commercial microwave. Histologically, periportal inflammation with heterophilic infiltration in the liver was noted. Spleens showed hyalinization of arteries, lymphoid depletion, and necrosis. Toe joints showed purulent synovitis and cellulitis. Gram stains done on impression smears of liver and spleen showed rare to moderate numbers of small gram-positive rods. Erysipelothrix rhusiopathiae was isolated from 18 of 22 livers cultured, five of six toe joints cultured, and from the yolk sac in two birds. PMID- 11007028 TI - Ornithobacterium rhinotracheale infection in commercial laying-type chickens. AB - Ornithobacterium rhinotracheale is a gram-negative, rod-shaped, pleomorphic bacterium that has been isolated from flocks of turkeys and broilers from around the world. Infections cause respiratory disease, mortality, and growth suppression, or clinical signs of infection may be absent. In layers, there have been few reports of disease caused by O. rhinotracheale. This is the first report of O. rhinotracheale infection in United States layer flocks. PMID- 11007029 TI - Turkey knockdown in successive flocks. AB - Turkey knockdown was diagnosed in three of five flocks of hen turkeys on a single farm within a 12-mo period. The age of birds in the flocks affected ranged from 6 wk 2 days to 7 wk 4 days. The attack rate ranged from 0.02% to 0.30% with a case fatality rate in affected birds ranging from 0 to 74%. The diagnosis was made on the basis of clinical signs and histopathologic lesions associated with knockdown. The feed in all flocks contained bacitracin methylene disalicylate and monensin (Coban). Affected birds were recumbent, demonstrated paresis, and were unable to vocalize. Postmortem examination revealed few significant lesions although pallor of the adductor muscles and petechiation in adductor and gastrocnemius muscles were noted. Birds that had been recumbent for extended periods were severely dehydrated. Consistent microscopic lesions included degeneration, necrosis, and regeneration of adductor, gastrocnemius, and abdominal muscles. No lesion in cardiac tissue was noted. Results of our investigation indicated that changes in water consumption, vitamin E status, and brooder to finisher movement correlated with the occurrence of knockdown. Turkey knockdown was defined in 1993 as any condition identified in a turkey flock that has affected the neuromuscular system to a degree that a turkey is unable to walk or stand. This definition was later modified to...neuromuscular or skeletal systems to a degree that a turkey is unable to walk or stand properly. Knockdown may be associated with numerous feed, management, or disease factors alone or in combination. Dosage of monensin, feed restriction/gorging, water restriction, heat stress, copper, mycotoxins, sodium chloride in feed, and sulfa drugs have all been suggested as contributing factors; however, laboratory studies to duplicate this have not been successful. This report presents observations from a single farm at which three of five hen flocks in a single year experienced knockdown. When a flock was reported as affected, a detailed investigation was initiated within 3 hr. The fifth flock was followed on a twice weekly basis from 0 to 8 wk of age to determine if initiating events were evident, but knockdown did not occur. PMID- 11007030 TI - Isolation of avian influenza virus (H10N7) from an emu (Dromaius novaehollandiae) with conjunctivitis and respiratory disease. AB - Avian influenza virus was isolated from the conjunctiva of a male emu chick. Clinical observations included ocular discharge, dyspnea, and mild respiratory signs. Lesions included conjunctivitis, tracheitis, bronchopneumonia, and airsacculitis. Escherichia coli was isolated from the conjunctiva and the sinus, and Staphylococcus sp. was isolated from the conjunctiva. Influenza A viral nucleoprotein was detected immunohistochemically in epithelial cells of the bronchi, lung parenchyma and tracheal mucosa, and mononuclear inflammatory cells within the exudate of the bronchial lumen; conjunctiva, air sacs, kidney, intestine, and liver were negative for the viral nucleoprotein. The isolated influenza virus was typed as H10N7 and was determined to be nonpathogenic for chickens. PMID- 11007031 TI - Expression of matrix metalloproteinase-1 in human colorectal carcinoma. AB - Matrix metalloproteinases are considered to play an important role in tumor invasion and metastasis. To elucidate the involvement of MMP-1 in human colorectal carcinoma, we performed immunohistochemical analysis on tissues from 20 colorectal adenomas and 142 colorectal adenocarcinomas, including 27 intramucosal carcinomas and 115 invasive carcinomas. MMP-1 was not expressed in any of the 20 cases of colorectal adenoma examined. In contrast, 108 of 142 cases (76.1%) with colorectal adenocarcinoma showed immunoreactivity for MMP-1 in the carcinoma cells themselves. Expression of MMP-1 was also identified in stromal cells around the carcinoma. We investigated the relationship between pathological features in colorectal carcinoma and MMP-1 immunoreactivity of the tumor cells. MMP-1 expression was less frequent in intramucosal carcinomas and weaker than that in invasive carcinomas (P < .0001). Among the 115 cases of invasive carcinomas, MMP-1 immunoreactivity was significantly correlated with the depth grading of tumor invasion (P < .05), tumor growth pattern (P < .05), the presence of lymphatic invasion (P < .05), venous invasion (P < .05), neural invasion (P < .05), lymph node metastasis (P < .005), hepatic metastasis (P < .05), and increasing stages of Dukes' classification (P < .05). In situ hybridization, using an MMP-1 oligonucleotide probe, confirmed the presence of MMP-1 mRNA in colorectal carcinoma cells themselves. Expression of MMP-1 mRNA was detected by the reverse transcription polymerase chain reaction method in cultured human colorectal carcinoma cell lines and colon carcinoma tissue obtained at surgery. These findings suggest that the expression of MMP-1 is one of the important factors related to tumor invasion and metastasis in colorectal carcinoma. PMID- 11007032 TI - Nodular amyloidoma and primary pulmonary lymphoma with amyloid production: a differential diagnostic problem. AB - Nodular amyloidomas (NA) of the lung are non-neoplastic inflammatory nodules containing eosinophilic amyloid deposits and a lymphoplasmacytic infiltrate. In some instances, the extensive amyloid deposits may obscure an underlying lymphoproliferative disorder. The histologic and immunohistologic features that discriminate these two differential diagnostic possibilities were studied in this series of six cases of NA and five cases of primary low-grade malignant lymphomas of lung with secondary amyloid deposits (ML). Two of lymphoma cases showed histopathologic and immunophenotypic features of B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-cell CLL/SLL), and three cases were low grade B-cell lymphoma derived from mucosa associated lymphoid tissue (MALT lymphoma). Key discriminating morphologic features between NA and ML included lymphatic tracking of the cellular infiltrate (3/5 ML; 1/6 NA), pleural infiltration (3/5 ML; 0/6 NA), sheet-like masses of plasma cells (5/5 ML; 0/6 NA) and reactive follicles (4/5 ML; 1/6 NA). Lesional circumscription, vascular and bronchial destruction, lymphoepithelial lesions, and granulomas were not helpful discriminators. Immunohistochemical features indicating a dominant CD20+, CD79a+ B-cell population (5/5 ML; 0/6 NA), light chain restriction (4/5 ML; 0/6 NA), and aberrant antigen expression of CD20/CD43 (2/5 ML; 0/6 NA) were helpful. Amyloid tumors with a reactive lymphoplasmacytic infiltrate can be separated from low grade malignant lymphomas utilizing both histologic and immunohistochemical features. PMID- 11007033 TI - What levels of agreement can be expected between histopathologists assigning cases to discrete nominal categories? A study of the diagnosis of hyperplastic and adenomatous colorectal polyps. AB - AIMS: To assess the levels of agreement between histopathologists for a two-class nominal categorization process--the discrimination between hyperplastic and adenomatous colorectal polyps. METHODS: Fifty hyperplastic and 50 adenomatous polyps received consecutively in the laboratory were categorized by nine histopathologists, and the level of agreement between all observers and the original diagnosis was assessed using kappa statistics. RESULTS: For the eight observers with 11 months or more experience in histopathology, there was a high level of agreement with kappa statistics ranging from 0.84 to 0.98. This process was performed rapidly with an average of 13 to 22 seconds spent on each case. One observer with only 6-weeks' experience of histopathology had a lower overall level of agreement with kappa statistics ranging from 0.46 to 0.54, but the performance on the later cases was much higher. CONCLUSIONS: The level of agreement in the distinction between hyperplastic and adenomatous colorectal polyps is high among histopathologists with at least moderate amounts of experience in histopathology. The one virtually naive observer showed a marked learning response during the study without feedback on case outcome. This suggests that histopathologists are very reliable in assigning cases to distinct nominal categories and that learning of these processes occurs early in a histopathologist's career. PMID- 11007034 TI - Abnormal expression of cell cycle regulatory proteins in ductal and lobular carcinomas of the breast. AB - In a previous study, we demonstrated that the G1 cell cycle checkpoint in carcinomas of the breast is frequently abrogated by loss of p16, the product of the CDKN2/INK4A gene, and, to a lesser extent, by loss of pRB, the product of the retinoblastoma gene. The purpose of the present study was to determine whether other mechanisms of cell cycle deregulation exist in breast cancers which have retained RB and p16 function. Paraffin sections of 81 invasive breast carcinomas (49 ductal, 26 lobular, 6 mixed) were reacted with monoclonal antibodies against cyclin D1 and p53, using optimized immunohistochemical staining protocols. The staining results were correlated with the expression of p16 and pRB, and with a variety of pathological parameters and DNA ploidy. Twenty-five tumors (31%) accumulated (presumably mutant) p53 and 28 (35%) overexpressed cyclin D1; 7 carcinomas (not including any pure lobular cancers) abnormally expressed both proteins. p53 accumulation correlated with nuclear, mitotic, and overall grade, but not with tumor size, lymph node involvement, or DNA ploidy. Overexpression of cyclin D1 was not associated with any of the patho-biological variables. There was an inverse correlation between loss of p16 and high levels of p53, but not cyclin D1. The G1 cell cycle checkpoint, which is controlled by RB, cyclin D1, and p16, was abrogated in 65% of carcinomas, and only p53 was abnormal in an additional 17%. The number of abnormally expressed genes correlated with mitotic activity and overall tumor grade, but not with tumor histology, size, or nodal status, suggesting that cell cycle deregulation is an early event in breast tumorigenesis. Only 18% of the carcinomas showed a normal level of expression of the four genes tested, and p16 appeared to be the most common target of cell cycle deregulation. These data point to the importance of cell cycle regulatory protein abnormalities in human breast cancer. PMID- 11007036 TI - Cytokeratin 7 and cytokeratin 20 expression in epithelial neoplasms: a survey of 435 cases. AB - Cytokeratin 7 (CK 7) and cytokeratin 20 (CK 20) are low molecular weight cytokeratins. Their anatomic distribution is generally restricted to epithelia and their neoplasms. We surveyed 435 epithelial neoplasms from various organ systems by immunohistochemistry using CK 7 and CK 20 monoclonal antibodies. Expression of CK 7 was seen in the majority of cases of carcinoma, with the exception of those carcinomas arising from the colon, prostate, kidney, and thymus; carcinoid tumors of the lung and gastrointestinal tract origin; and Merkel cell tumor of the skin. The majority of cases of squamous cell carcinoma of various origins were negative for CK 7, except cervical squamous cell carcinoma, in which 87% of cases were positive. Approximately two thirds of cases of malignant mesothelioma were CK 7-positive. CK 20 positivity was seen in virtually all cases of colorectal carcinomas and Merkel cell tumors. CK 20 positive staining was also observed in cases of pancreatic carcinomas (62%), gastric carcinoma (50%), cholangiocarcinomas (43%), and transitional cell carcinomas (29%). The expression of CK 20 was virtually absent in carcinomas from other organ systems and in malignant mesothelioma. CK 7- and CK 20-negative epithelial neoplasms included adrenal cortical carcinoma, germ cell tumor, prostate carcinoma, renal cell carcinoma, and hepatocellular carcinoma. PMID- 11007035 TI - Pathologic, cytogenetic and molecular assessment of acute promyelocytic leukemia patients treated with arsenic trioxide (As2O3). AB - Arsenic trioxide (As2O3) shows great promise as an effective therapy for patients with all-trans retinoic acid (ATRA)-resistant acute promyelocytic leukemia (APL). Little data is available addressing the pathology of As2O3 treated APL and whether the antileukemic mechanism of As2O3 is primarily cytolysis or through stimulation of cell differentiation. In this report, we made a morphologic, cytogenetic, and molecular evaluation of five ATRA-refractory APL patients who were treated with As2O3. Four of the five patients had morphologic responses after one or two cycles of As2O3 treatment. Of the four responders based on bone marrow morphology, two achieved molecular remission (negative RT-PCR for PML- RAR alpha fusion transcripts) by the end of the second and third cycles of As2O3 therapy. Two patients exhibited marked leukocytosis during the first cycle of As2O3, and at that time point the APL cells were largely replaced by the cells showing partial differentiation towards myelocytes with co-expression of CD11b and CD33. Nevertheless, these "myelocyte-like" cells that showed the t(15;17) translocation eventually disappeared with continuous As2O3 therapy. As2O3 treatment appears to be effective therapy for the patients with relapsed APL after the failure of conventional chemotherapy and ATRA therapy. The pathologic findings in these five cases suggest that at low doses As2O3 primarily induces differentiation of the APL cells, generating abnormal myelocytes resembling APL cells treated with ATRA, whereas at higher doses AS2O3 induces marrow necrosis. PMID- 11007037 TI - Missense mutation of the MET gene detected in human glioma. AB - Multiple mechanisms, such as gene mutations, amplifications, and rearrangements, as well as perturbed mitogen and receptor function, are likely to contribute to glioma formation. The MET (also known as c-met proto-oncogene located at 7q31-34 has been shown to be amplified in human gliomas, and activating mutations within the tyrosine kinase domain of MET have been causally related to tumorigenesis in hereditary papillary renal cell carcinoma. To elucidate the role of MET gene in glioma formation, sporadic gliomas from 11 patients were examined for MET gene mutations and allelic duplications or deletions by polymerase chain reaction single strand conformational polymorphism analysis and fluorescence in situ hybridization. Three of 11 sporadic gliomas showed a deletion of one copy of the MET gene, and a specific METgene missense mutation in the remaining gene copy was detected in one of those tumors. The corresponding sequence in non-tumor DNA was normal in all cases. Three of 11 sporadic gliomas showed duplication of one copy of the MET gene, but none of them contained mutations. One tumor showed METamplification without mutation. Three showed neither allelic change nor mutation. These data suggest that somatic MET gene mutation may play a role in the development of a subgroup of sporadic gliomas. However, MET mutations appear to be absent in the majority of sporadic gliomas. PMID- 11007038 TI - Splenic angiosarcoma: a clinicopathologic and immunophenotypic study of 28 cases. AB - Primary angiosarcoma of the spleen is a rare neoplasm that has not been well characterized. We describe the clinical, morphologic, and immunophenotypic findings of 28 cases of primary splenic angiosarcoma, including one case that shares features of lymphangioma/lymphangiosarcoma. The patients included 16 men and 12 women, aged 29 to 85 years, with a mean of 59 years and median of 63 years. The majority of patients (75%) complained of abdominal pain, and 25% presented with splenic rupture. The most common physical finding was splenomegaly (71%). Seventeen of 21 patients were reported to have anemia. Macroscopic examination showed splenomegaly in 85% cases. Sectioning revealed discrete lesions in 88% of cases, ranging from well-circumscribed firm nodules to poorly delineated foci of necrosis and hemorrhage associated with cystic spaces. Microscopically, the tumors were heterogenous; however, all cases demonstrated at least a focal vasoformative component lined by atypical endothelial cells. Solid sarcomatous, papillary, and epithelioid growth patterns were observed. The solid sarcomatous component resembled fibrosarcoma in two cases and malignant fibroushistiocytoma in one case. Hemorrhage, necrosis, hemosiderin, extramedullary hematopoiesis, and intracytoplasmic hyaline globules were frequently identified. A panel of immunohistochemical studies revealed that the majority of tumors were immunoreactive for at least two markers of vascular differentiation (CD34, FVIIIRAg, VEGFR3, and CD31) and at least one marker of histiocytic differentiation (CD68 and/or lysozyme). Metastases developed in 100% of patients during the course of their disease. Twenty-six patients died of disease despite aggressive therapy, whereas only two patients are alive at last follow-up, one with disease at 8 years and the other without disease at 10 years. In conclusion, primary splenic angiosarcoma is an extremely aggressive neoplasm that is almost universally fatal. The majority of splenic angiosarcomas coexpress histiocytic and endothelial markers by immunohistochemical analysis, which suggest that some tumors may originate from splenic lining cells. PMID- 11007039 TI - Myogenin is a specific marker for rhabdomyosarcoma: an immunohistochemical study in paraffin-embedded tissues. AB - Myogenin belongs to a group of myogenic regulatory proteins whose expression determines commitment and differentiation of primitive mesenchymal cells into skeletal muscle. The expression of myogenin has been demonstrated to be extremely specific for rhabdomyoblastic differentiation, which makes it a useful marker in the differential diagnosis of rhabdomyosarcomas (RMS) from other malignant small round cell tumors of childhood. Commercially available antibodies capable of detecting myogenin in routinely processed formalin-fixed paraffin-embedded (FFPE) tissue are now available. In this study, we evaluated myogenin expression using the monoclonal myf-4 antibody (Novocastra Labs) on FFPE in a large number of pediatric tumors in order to define the clinical utility of this marker. A total of 119 tumors were studied. These included 48 alveolar RMS (ARMS), 20 embryonal RMS (ERMS), one spindle cell RMS, 16 Ewing's sarcomas (ES), six nephroblastomas, two ectomesenchymomas, seven precursor hematopoietic neoplasms, five olfactory neuroblastomas, three neuroblastomas, six desmoplastic small round cell tumors, and five rhabdoid tumors. Distinct nuclear staining for myogenin was noted in all 69 RMS. Notably, the number of positive tumor cells differed between the ARMS and ERMS. In ARMS, the majority of tumor cells (75 to 100%) were positive, in contrast to ERMS, in which the positivity ranged from rare + to 25% in all but three tumors. Additionally, myogenin positivity was seen in two of two ectomesenchymomas and in two nephroblastomas with myogenous differentiation. All other tumors were clearly negative. Our results indicate that staining for myogenin is an extremely reliable and specific marker for rhabdomyoblastic differentiation. It gives consistent and easily interpretable results in routinely fixed tissues. PMID- 11007040 TI - Molecular abnormalities of p53, MDM2, and H-ras in synovial sarcoma. AB - Forty-nine cases of synovial sarcoma were evaluated for mutation of the p53 gene, amplification of the MDM2 gene and mutation of the H-ras gene, and for the relation of these factors to overall survival and clinicopathologic parameters. All investigations were carried out on formalin-fixed paraffin-embedded materials. Furthermore, we evaluated the expression of p53 protein, MDM2, and p21(WAF1/CIP1) immunohistochemically in these cases, together with an assessment of proliferative activities using monoclonal antibody MIB-1. Nine of the 49 cases (18.4%) had p53 gene alteration detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and direct sequencing. Eleven cases (24%) showed nuclear accumulation of p53 protein in more than 10% of the tumor cells. Among them, only three cases contained gene mutations. There was no correlation between p53 nuclear accumulation and p53 gene alteration. MDM2 gene amplification, as shown by differential PCR, was observed in 19 out of 47 cases (40%). Nineteen out of 49 cases (38.8%) showed immunoreactivity for MDM2. MDM2 gene amplification and the expression of MDM2 protein showed a significant positive relationship (P = 0.0004). Moreover, MDM2 immunoreaction was significantly correlated with nuclear accumulation of p53 protein (P = 0.023). Positive immunoreaction for p21(WAF1/CIP1) was observed in 21 out of 48 cases (43.8%). p21(WAF1/CIP1) expression was correlated with p53 protein expression. H ras gene mutations were seen in only three cases (6.1%). All mutations were in codon 12 (one GGC-to-AGC [Gly-to-Ser] mutation and two GGC-to-GAC [Gly-to-Ap] mutations). The gene alteration of p53, MDM2, and H-ras did not affect the patients' prognosis. Although the cases with positive immunoreaction for p53 tended to have a worse prognosis, the difference was not statistically significant (P = 0.13). No correlation was observed between MIB-1 LI and the immunohistochemical expression of p53, MDM2, and p21(WAF1/CIP1) or the mutation status of p53 and H-ras. On the other hand, high MIB-1 LI (more than 10) significantly correlated with poor prognosis (P < 0.0001). Our results suggest that p53 gene mutation does not appear to be a major prognostic factor and H-ras mutations are infrequent in synovial sarcoma. PMID- 11007041 TI - Beta-catenin in soft tissue sarcomas: expression is related to proliferative activity in high-grade sarcomas. AB - Besides its role in cell adhesion, beta-catenin exerts a function as an oncoprotein. The aim of this study was the characterization of its expression, possible mutation, and the assessment of beta-catenin as a prognostic indicator for soft tissue sarcomas. A total of 115 soft tissue sarcomas were analyzed using immunohistochemistry, immunogold-electron microscopy, and DNA analysis. Information from 56 patients was available for follow-up. A statistically significant correlation was found between intracellular distribution of beta catenin and the proliferative activity (MIB-1 expression) in high-grade sarcomas (P = .0008). Beta-catenin was identified with intracytoplasmic and nuclear accumulation, showing additional membranous staining in sarcomas with epithelioid pattern. Ultrastructurally, a colocalization between beta-catenin and nuclear heterochromatin was demonstrated. In 22 analyzed tumors, only one (yet undescribed) mutation of the beta-catenin gene (C-A transversion) could be detected. Prognostic validity of the cellular expression of beta-catenin, however, was not proven. Apart from its membranous function as an effective molecule for cell-adhesion in sarcomas with epithelioid pattern, beta-catenin may act as an oncoprotein in sarcomas with intracytoplasmic and nuclear localization with binding to nuclear DNA. A previously discussed stimulation of cell proliferation caused by an increased beta-catenin level can also be postulated for high-grade soft tissue sarcomas in correlation with the rate of proliferation. Mutations of the beta-catenin gene are probably of lesser importance for the accumulation of beta-catenin in soft tissue sarcomas. PMID- 11007042 TI - p27kip1 expression distinguishes papillary hyperplasia in Graves' disease from papillary thyroid carcinoma. AB - In most cases, the histopathologic and cytologic distinction between Graves' disease and papillary thyroid carcinoma is relatively easy, but on occasion Graves' disease may simulate a thyroid papillary carcinoma. For example, papillary fronds with fibrovascular cores may be present in both Graves' disease and papillary carcinoma. p27kip1 (p27) is a cyclin-dependent kinase inhibitory protein that has been shown to be an independent prognostic factor in a variety of human tumors. Our previous studies of p27 expression in hyperplastic and neoplastic endocrine lesions showed that the level of p27 was quite different in these two conditions. To determine if this distinction could also be made between Graves' disease and papillary carcinoma, we analyzed expression of p27 and other cell cycle proteins in a series of cases of Graves' disease with papillary hyperplasia and a series of papillary thyroid carcinomas. Formalin-fixed paraffin embedded tissues from 61 randomly selected patients with thyroid disease, including 29 cases of Graves' disease with papillary architectural features and 32 cases of papillary carcinoma, were analyzed for expression of p27, Ki-67, and DNA topoisomerase II alpha (topo II alpha) by immunostaining. The distribution of immunoreactivity was analyzed by quantifying the percentage of positive nuclei that was expressed as the labeling index (LI) plus or minus the standard error of the mean. The papillary hyperplasia of Graves' disease had a p27 LI of 68.2 +/- 3.1 (range, 24 to 88), whereas papillary carcinomas had a LI of 25.6 +/- 2.5 (range, 12 to 70) (P < .0001). No significant differences in Ki-67 or topo II alpha expression were identified between papillary hyperplasia in Graves' disease and papillary carcinoma. These results indicate that p27 protein expression is significantly higher in papillary hyperplasia of Graves' disease compared to papillary carcinoma, which may be diagnostically useful in difficult cases. PMID- 11007044 TI - Recommendations for the reporting of surgical specimens containing uterine cervical neoplasms. PMID- 11007043 TI - Increased density of interstitial mast cells in amyloid A renal amyloidosis. AB - Renal interstitial fibrosis is the final common pathway leading to end-stage renal disease in various nephropathies including renal amyloidosis. However, the role of mast cells (MCs) in the fibrotic process of renal amyloidosis is not fully understood. We compared the distribution of MCs in renal biopsies from 30 patients with AA type renal amyloidosis and 20 control cases. Immunoreactivity of renal MCs to anti-tryptase and anti-chymase was studied. Interstitial myofibroblasts were stained with anti-alpha-smooth muscle actin (alpha-SMA) antibody, and inflammatory cells were identified by anti-CD45, -CD20, and -CD68 mAbs. Positively stained cells were counted, and the relative interstitial and fractional areas of anti-alpha-SMA stained cells were measured. Anti-CD29 mAb was used to detect beta1 integrin and anti-basic fibroblast growth factor (bFGF) mAb for the growth factor on MCs. MCs were rarely found in control samples. In contrast, samples showing amyloid deposition contained numerous tryptase-positive (MCT) (940.17 +/- 5.4 versus 6.74 +/- 1.1/mm2) but fewer chymase-positive (MCTC) cells (20.7 +/- 2.86 versus 1.7 +/- 0.76/mm2) in the renal interstitium. There was a significant relationship between interstitial MCT and creatinine clearance (r = -0.72), and between interstitial MCT and glomerular amyloid-index (GAI) (r = 0.723) and interstitial amyloid area (r = 0.824). Accumulation of MCs correlated significantly with the number of T lymphocytes (MCT: r = 0.694). There was also a significant relationship between mast cell (MC) number and the fractional area of alpha-SMA positive interstitium (r = 0.733) and interstitial fibrotic area (r = 0.6). Double immunostaining demonstrated intracytoplasmic presence of beta1 integrin on 87% of MCT and correlated significantly with the interstitial amyloid area (r = 0.818, P = .001) and T-cell number (r = 0.639, P = .002). bFGF was also detected on 85.5% of MCTC correlating well with the interstitial alpha-SMA-area (r = 0.789). Our results indicate that MCs constitute an integral part of the overall inflammatory process and play a crucial role in interstitial fibrosis in renal amyloidosis. PMID- 11007045 TI - Recommendations for the reporting of resected esophageal carcinomas. PMID- 11007046 TI - Recommendations for the reporting of specimens containing oral cavity and oropharynx neoplasms. PMID- 11007047 TI - Recommended reporting format for thyroid carcinoma. PMID- 11007048 TI - Protocol for malignant and potentially malignant neoplasms of the testis and paratestis. PMID- 11007049 TI - The importance of documenting the appearance and status of breast implants at time of explantation. PMID- 11007050 TI - Acute clinical events in 299 homozygous sickle cell patients living in France. French Study Group on Sickle Cell Disease. AB - A subset of 299 patients with homozygous sickle cell anaemia, enrolled in the cohort of the French Study Group on sickle cell disease (SCD), was investigated in this study. The majority of patients were children (mean age 10.1 +/- 5.8 yr) of first generation immigrants from Western and Central Africa, the others originated from the French West Indies (20.2%). We report the frequency of the main clinical events (mean follow-up 4.2 +/- 2.2 yr). The prevalence of meningitis-septicaemia and osteomyelitis was, respectively, 11.4% and 12% acute chest syndrome was observed in 134 patients (44.8%). Twenty patients (6.7%) developed stroke with peak prevalence at 10-15 yr of age. One hundred and seventy two patients (58%) suffered from one or more painful sickle cell crises, while the others (42.5%) never suffered from pain. The overall frequency of acute anaemic episodes was 50.5%, (acute aplastic anaemia 46%; acute splenic sequestration 26%). A group of 27 patients were asymptomatic (follow-up > 3 yr). Epistatic mechanisms influencing SCD were studied. Coinherited alpha-thalassemia strongly reduced the risk of stroke (p <0.001) and increased that of painful crises (p < 0.02). There was a low prevalence of Senegal and Bantu (CAR) betas chromosomes in patients with meningitis (p <0.04) and osteomyelitis (p < 0.03). Prevalence of Senegal betas-chromosomes was lower in the asymptomatic group of 27 patients (p < 0.02). The patients come from a population of unmixed immigrants in whom the beta-globin gene haplotype strongly reflects the geographic origin and identifies subgroups with a homogenous genetic background. Thus the observed effects might result more from differences in as yet unidentified determinants in the genetic background than from the direct linkage with differences in the beta globin gene locus. PMID- 11007051 TI - Pregnancy in essential thrombocythaemia: treatment and outcome of 17 pregnancies. AB - OBJECTIVE: To evaluate treatment and outcome of 17 pregnancies in nine patients with essential thrombocythaemia (ET) seen at our institution from 1988 to 1998. METHODS: Treatment and outcome of 17 pregnancies in nine ET patients were retrospectively analyzed. RESULTS: Seventeen pregnancies in nine patients with ET resulted in 11 (65%) live births and ended in six (35%) spontaneous abortions. Abortion could not be predicted from ET-associated complications before (p= 0.23) or during (p = 0.39) pregnancy. Maternal complications occurred during six pregnancies (35%): Three major bleedings in two patients with an acquired von Willebrand disease and two minor bleedings in patients treated with low-dose acetylsalicylic acid (ASA) were observed during pregnancy or at term; one patient suffered from transient visual loss while pausing low-dose ASA. Platelet counts prior to pregnancy were significantly higher as compared to the platelet nadir observed during pregnancy (p = 0.0017). Postpartum clinical course was uneventful in all patients. No specific treatment was given during 11 pregnancies. Six women received low-dose ASA during pregnancy followed by low-molecular-weight heparin until the end of the sixth week postpartum in five cases. This treatment was correlated with a favourable outcome (live birth versus abortion) when compared to no treatment (p=0.04). CONCLUSION: Pregnancy in ET can be complicated by first trimester abortion and/or maternal haemorrhage. Our limited observation suggest a positive impact of low-dose ASA during pregnancy followed by low-molecular-weight heparin postpartum on pregnancy outcome in ET; nevertheless, confirmation by prospective documentation is mandatory. PMID- 11007052 TI - Megakaryocyte c-Mpl expression in chronic myeloproliferative disorders and the myelodysplastic syndrome: immunoperoxidase staining patterns and clinical correlates. AB - The objectives of this study were to expand on recent observations that have suggested decreased thrombopoietin receptor (c-Mpl) expression in megakaryocytes of patients with polycythemia vera (PV) and agnogenic myeloid metaplasia (AMM). We applied an immunoperoxidase method with anti-c-Mpl antibody to 55 bone marrow sections from previously untreated patients with chronic myeloproliferative disorder (CMPD) or myelodysplastic syndrome (MDS). These included 8 patients with PV, 15 with AMM, 9 with essential thrombocythemia, 5 with chronic myelocytic leukemia, 9 with the 5q-syndrome and 9 with MDS with fibrosis. The findings were compared with those in four patients with reactive erythrocytosis (RE), six with immune thrombocytopenic purpura (ITP) and five normal controls. Staining intensity (SI) was moderate to strong both in normal controls and in patients with RE or ITP. In contrast, SI was weak in variable proportions of the megakaryocytes in every one of the aforementioned clonal myeloid disorders. The staining pattern (SP) was relatively uniform in MDS and heterogeneous in CMPD. Neither SI nor SP was significantly correlated with certain clinical or laboratory parameters. We concluded that altered megakaryocyte c-Mpl expression may be a nonspecific phenomenon in various subtypes of both CMPD and MDS. However, the characteristic staining patterns may complement the morphological distinction between clonal and reactive myeloproliferation. PMID- 11007053 TI - Renal failure in multiple myeloma: reversibility and impact on the prognosis. Nordic Myeloma Study Group. AB - The purpose of the present study was to analyse the importance and prognostic value of renal failure in multiple myeloma patients. The frequency and reversibility of renal failure in 775 multiple myeloma patients diagnosed between 1984-86 and 1990-92 in the Nordic countries were studied. Renal failure, defined as plasma creatinine > 130 micromol/l, was observed in 29% of the cases at the time of diagnosis. During the first year after diagnosis 58% achieved normalisation of p-creatinine, and this was achieved mainly during the first 3 months. Reversibility of renal failure was more frequently observed in patients with moderate renal failure, hypercalcaemia and low Bence-Jones protein excretion. In a multivariate analysis renal failure, high age, stage III disease and hypercalcaemia were independent prognostic factors for survival. Patients who needed dialysis had a poor prognosis, with a median survival of 3.5 months. A 12 months landmark analysis showed that reversibility of renal failure was a more important prognostic factor than response to chemotherapy. It is concluded that renal failure in multiple myeloma is reversible in about half the cases, and reversibility of renal failure improves long-term survival. PMID- 11007054 TI - Triple Philadelphia chromosomes with major-bcr rearrangement in hypotriploid erythroleukaemia. AB - The Philadelphia (Ph) chromosome is observed in approximately 1% of patients with acute myeloblastic leukaemia (AML), especially subtypes M1 and M2 in the French American-British classification. We describe here a cytogenetic and molecular investigation of a rare case with Ph-positive AML M6 (erythroleukaemia). A 63-yr old woman was diagnosed as having erythroleukaemia. Leukaemic cells were positive for CD4 and CD7 as well as CD13, CD33, CD34 and HLA-DR. They were analyzed by G banding, fluorescence in situ hybridization (FISH), Southern blot and reverse transcriptase polymerase chain reaction analyses. The karyotypes at diagnosis were as follows: 61, XX, -X, -1, -2, -3, -4, -5, -7, t(9;22)(q34;q11)x 2, -15, 16, -17, -18, + 19, +21, +22 [3]/61, idem, -22, +der(22)t(9;22) [36]. FISH with BCR/ABL probes showed that 39% and 57% of interphase nuclei had double and triple BCR/ABL fusion signals, respectively. Chromosome analysis in complete remission showed a normal karyotype in all 20 metaphases, confirming the diagnosis as Ph positive-acute leukaemia. FISH at relapse showed that 92% of interphase nuclei had triple fusion signals. Rearrangement of major breakpoint cluster region (M bcr) in the BCR gene and coexpression of p210-type (b2a2) and p190-type (e1a2) BCR/ABL fusion transcripts due to alternative splicing were also detected. We conclude that clonal evolution from double to triple Ph chromosomes may be implicated in the disease progression. Considering other two reported cases, Ph positive erythroleukaemia appears to be correlated with coexpression of myeloid/T lymphoid markers and hyperdiploidy with double or triple Ph chromosomes, although breakpoints in the BCR gene are heterogenous. PMID- 11007055 TI - CyclOBEAP (cyclophosphamide, vincristine, bleomycin, etoposide, doxorubicin, prednisolone) regimen with granulocyte colony-stimulating factor (G-CSF) for patients with aggressive non-Hodgkin's lymphoma: a pilot study. The Adult Lymphoma Treatment Study Group (ALTSG). AB - We conducted a multi-institutional collaborative study to examine the usefulness and safety of third-generation chemotherapy CyclOBEAP (cyclophosphamide, vincristine, bleomycin, etoposide, doxorubicin, prednisolone) combined with granulocyte colony-stimulating factor (G-CSF) in the treatment of aggressive non Hodgkin's lymphoma (NHL). Subjects included patients with aggressive NHL who were 60 yr of age or younger and had been diagnosed as having a low-intermediate, high intermediate, or high risk using the International Prognostic Index (IPI). A total of 24 patients were enrolled in the study between May 1997 and March 1998, including 9 low-intermediate-risk cases, 13 high-intermediate-risk cases and 2 high-risk cases. Although all 24 patients were originally enrolled in the study, one adult T-cell leukemia/lymphoma case was subsequently excluded. Thus, in the end, 23 cases were evaluated. Evaluation of the efficacy of therapy revealed complete remission in 20 patients (87%). Of these 20 patients, 8 were low intermediate-risk cases (89%) and 12 were either high-intermediate- or high-risk cases (86%). Partial remission was achieved in 2 patients (8.7%). The 2-yr survival rate was 91.3%, and the 2-yr disease-free survival rate was 81.8%. Grade 3 or higher adverse reactions were granulocytopenia (87%), thrombocytopenia (17.4%) and liver dysfunction (4.3%). CyclOBEAP therapy has been associated with a high remission rate for aggressive NHL. When combined with G-CSF, a high relative dose intensity was maintained for each drug administered (0.94-0.97). Furthermore, although the observation period was short, both the survival rate and disease-free survival rate were good. Hence, we concluded that there were no problems associated with the procedure in terms of safety. PMID- 11007056 TI - Elevated serum levels of soluble CD44 variant 6 are correlated with shorter survival in aggressive non-Hodgkin's lymphoma. AB - A variant form of CD44 that has additional amino acids in the common protein backbone (CD44-v6) seems to play a role in the metastasis of malignancies. We measured soluble CD44-v6 (sCD44-v6) by ELISA in 201 patients with malignant lymphoma. The sCD44-v6 level was significantly elevated in patients with non Hodgkin's lymphoma (NHL) (n = 184). The sCD44-v6 level was correlated significantly with the standard sCD44 and soluble interleukin-2 receptor levels, but only weakly with serum lactate dehydrogenase (LDH). In 149 patients with aggressive NHL, the sCD44-v6 level was elevated in the subgroups with a high LDH level, stage III/IV disease, T-cell lymphoma, and high-intermediate or high risk group as identified by the International Prognostic Index (IPI). When the sCD44 v6 level was > or = 800 ng/ml the overall survival rate was significantly decreased (p = 0.0001). In the low + low-intermediate risk group (IPI) both overall survival rates (log-rank p = 0.0005, Wilcoxon p =0.002) were significantly decreased when the sCD44-v6 level was > or = 800 ng/ml. In multivariate analysis, sCD44-v6 was shown to be independent of the five prognostic factors in the IPI (age, performance status, number of extranodal sites, Ann Arbor stage and LDH level), so it may be useful for predicting the outcome of aggressive NHL. PMID- 11007057 TI - Molecular and cytogenetic remission in a case of subtype M4E acute myelogenous leukemia with minimal monochemotherapy: high sensitivity or spontaneous remission? AB - Complete remission was observed in an adult patient with acute myelogenous leukemia after minimal monochemotherapy. Remission occurred after a severe febrile pneumonia and was accompanied by cytogenetic and molecular remission. The hypothesis of spontaneous remission was raised, even if a high sensitivity to low dose cytostatics cannot be excluded. Such spontaneous complete remissions, often associated with bacterial infections and blood transfusions, are extremely rare, and are usually of short duration. Previous cases are summarized, and the role of etiologic factors is discussed. PMID- 11007058 TI - Interferon alpha is an effective therapy for congenital dyserythropoietic anaemia type I. AB - The efficacy of interferon-alpha (IFN) was reported in three patients with congenital dyserythropoietic anaemia (CDA) type I. We describe two additional cases treated with IFN, which normalized the haemoglobin level in both patients with a dramatic decrease in the size of the spleen in one. Haemoglobin remained stable more than 6 months after discontinuation of treatment. IFN induced more than 50% decrease in the number of BFU-E in both patients' bone marrow cultures, suggesting an indirect effect of IFN on erythropoiesis in vivo. We conclude that a trial of IFN therapy should be considered in patients with CDA type I. PMID- 11007059 TI - The pathogenetic mechanism of myeloid malignancies associated with deletions of the long arm of chromosome 20 can not be explained by a "one hit" model. An acute myeloid leukemia patient who developed with 20q- clone during complete remission for 9 years. PMID- 11007060 TI - Chronopharmaceutical drug delivery from a pulsatile capsule device based on programmable erosion. AB - We report the development of a chronopharmaceutical capsule drug delivery system capable of releasing drug after pre-determined time delays. The drug formulation is sealed inside the insoluble capsule body by an erodible tablet (ET). The release time is determined by ET erosion rate and increases as the content of an insoluble excipient (dibasic calcium phosphate) and of gel-forming excipient (hydroxypropylmethylcellulose; HPMC) increases. The time-delayed release of a model drug (propranolol HCI) was investigated by dissolution testing (USP XXIII paddle method). Both composition and weight of ET influence the time of drug release. Moreover it was found that drug release was controlled by the quantity of HPMC, irrespective of lactose content within the tablet weight range 80-160 mg, when above a threshold concentration of 20% HPMC. Programmable pulsatile release has been achieved from a capsule device over a 2-12-h period, consistent with the demands of chronotherapeutic drug delivery. The time of drug release can be controlled by manipulation of tablet formulation. PMID- 11007061 TI - Improvement of subcutaneous bioavailability of insulin by sulphobutyl ether beta cyclodextrin in rats. AB - The objective of this study was to examine and compare how hydrophilic beta cyclodextrin derivatives (beta-CyDs) improve the bioavailability of insulin following subcutaneous injection of insulin solution in rats. When insulin solutions in the absence of beta-CyDs were injected into the dorsal subcutaneous tissues of rats, the absolute bioavailability of insulin calculated from plasma immunoreactive insulin (IRI) levels was approximately 50%. When maltosyl-beta cyclodextrin was added to the solutions, there was no change in the plasma IRI levels and hypoglycaemia compared with those of the insulin-alone solution. Dimethyl-beta-cyclodextrin decreased the bioavailability of insulin, although it increased the maximal concentration of IRI in plasma and the capillary permeability of the fluorescein isothiocyanatedextran 40, a non-degraded permeation marker. When insulin solutions containing sulphobutyl ether-beta cyclodextrin with a degree of substitution of the sulphobutyl group of 3.9 (SBE4 beta-CyD) were injected, the IRI level rapidly increased and maintained higher IRI levels for at least 8 h. The bioavailability of the insulin/SBE4-beta-CyD system was about twice that of insulin alone and approached 96%. The enhancing effects of SBE4-beta-CyD may be in part due to the inhibitory effects of SBE4 beta-CyDs on the enzymatic degradation and/or the adsorption of insulin onto the subcutaneous tissue at the injection site, although this does not apparently facilitate capillary permeability. These results suggest that SBE4-beta-CyD in aqueous insulin injection for subcutaneous administration is useful for improving the bioavailability and the hence the pharmacological effects of insulin. PMID- 11007062 TI - Ion-pair formation as a strategy to enhance topical delivery of salicylic acid. AB - An in-vitro study was carried out to determine the possibility of improving the efficiency of transdermal delivery of salicylate through human epidermis by ion pair formers (alkylamines and quaternary ammonium ions). Further, the relationship between the physicochemical properties of the counter-ions and salicylate flux was examined. It was found that flux can be related to the conductivity associated with the penetrant solution, molecular size of the counter-ion and lipophilicity expressed as either octanol/water partition coefficient of the ion pairs or the carbon chain-length of the counter-ions. Equations have been developed to predict salicylate flux from these physicochemical parameters. PMID- 11007064 TI - An investigation into the low temperature thermal behaviour of vitamin E preparation USP using differential scanning calorimetry and low frequency dielectric analysis. AB - The thermal and dielectric responses of Vitamin E Preparation USP have been examined to further understand the melting and solidification of this material. A TA Instruments 2920 Differential Scanning Calorimeter was used to examine the thermal response of the sample at a range of scanning speeds. Isothermal dielectric studies were performed using a Novocontrol Dielectric Spectrometer over a range of temperatures down to -70 degrees C and a frequency range of 10(6) 10(-2) Hz. The differential scanning calorimetry (DSC) studies showed an anomalous response whereby at slow heating rates (2 degrees C min(-1)) a small exotherm followed immediately by an endotherm was observed. This response was considerably diminished in magnitude at higher rates (5 degrees C min(-1)) and was not observed at the fastest heating rate of 10 degrees C min(-1). No thermal events were seen on cooling the sample to -60 degrees C. It was suggested that the material formed a glass on cooling, with a predicted transition temperature of approximately -100 degrees C. Further studies using a liquid nitrogen cooling system indicated that the system did indeed exhibit a glass transition, albeit at a higher temperature than predicted (ca -63 degrees C). Low frequency dielectric analysis showed a clear relaxation peak in the loss component, from which the relaxation time could be calculated using the Havriliak-Negami model. The relationship between the relaxation time and the temperature was studied and was found to follow the Vogel-Tammann-Fulcher (VTF) modification of the Arrhenius equation. It is therefore concluded that Vitamin E Preparation USP is a glass forming material that exhibits kinetically-hindered recrystallisation and melting behaviour. The study has also indicated that DSC and low frequency dielectric analysis may be powerful complementary tools in the study of the low temperature behaviour of pharmaceuticals. PMID- 11007063 TI - Effect of ion pairing with alkylamines on the in-vitro dermal penetration and local tissue disposition of salicylates. AB - Hydrophilic ionic drugs can be rendered lipophilic by ion-pair formation with hydrophobic counter-ions. This study examines the value of forming ion pairs between anionic salicylate and a series of amines as model cationic counter-ions to facilitate topical delivery and skin penetration. The in-vitro translocation of salicylate ions from a nonaqueous vehicle through human epidermis was estimated in the presence or absence of amines. The distribution into, and accumulation of the salicylate ion in various tissues following topical application to anaesthetised rats were also investigated. Although the epidermal permeation constants of the salicylate-amine ion pairs were lower than that of salicylate itself (enhancement ratios: 0.74-0.87), salicylate retention and localisation in the underlying rat tissues increased in the presence of some of the counter-ions studied. Salicylate concentrations (microg (g tissue)(-1)) in the dermis were 877.2+/-78.6 for salicylate alone and 1098+/-121.9-2586+/-332.5 for salicylate-amine ion pairs. The levels of salicylate in tissues up to the top muscle layer were 1.2-3.7-fold higher in the presence of the counter-ions. It is concluded that, although amine counter-ions have the ability to influence the penetration of salicylate, in-vitro permeability studies do not reflect the in vivo increases in tissue concentrations resulting from probable changes in systemic clearance. PMID- 11007065 TI - Solid dispersion of hydroxypropyl beta-cyclodextrin and ketorolac: enhancement of in-vitro dissolution rates, improvement in anti-inflammatory activity and reduction in ulcerogenicity in rats. AB - Ketorolac, is a non-steroidal anti-inflammatory drug, with strong analgesic activity. It is practically insoluble in water and has been implicated in causing gastrointestinal ulceration. This study describes the formulation of solid dispersions of ketorolac using hydroxypropyl beta-cyclodextrin (HPbeta-CyD) and beta-cyclodextin (beta-CyD) as carriers, to improve the aqueous solubility of the drug, thus enhancing its bioavailability. Also, reduction in ulcerogenicity was anticipated. Differential scanning calorimetry and X-ray diffraction studies indicated loss of crystalline nature of the drug, in the dispersions prepared with HPbeta-CyD. NMR studies revealed a strong interaction between drug and HPbeta-CyD. Solid dispersions of drug with beta-CyD retained the crystalline nature of the drug. All the solid dispersions showed a remarkable improvement in the rate and extent of dissolution of ketorolac. The kneaded dispersion with HPbeta-CyD prepared using a 1:1 alcohol-water mixture showed promise in reducing the ulcer-inducing effect of ketorolac in rats. Oral administration of this dispersion was found to inhibit carrageenan-induced paw oedema in rats to a significantly greater extent compared with ketorolac or its trometamol salt. Though beta-CyD as a carrier for ketorolac gave faster release of the poorly soluble drug, HPbeta-CyD proved to be superior to beta-CyD, as a carrier in the kneaded dispersion prepared using 1:1 alcohol-water mixture. These results suggest that solid dispersions of ketorolac with HPbeta-CyD aid in faster dissolution and better bioavailability of the drug. The higher solubility of the drug in the presence of HPbeta-CyD also reduces local gastrointestinal side effects of the drug. PMID- 11007066 TI - Estimation of oral bioavailability in the rat by the accelerated infusion technique. PMID- 11007067 TI - Simultaneous determination of unbound cefoperazone in rat blood and brain using microdialysis. AB - A sensitive microbore HPLC method was developed for the simultaneous determination of unbound cefoperazone in rat blood and brain using microdialysis. Two microdialysis probes were inserted into the jugular vein/right atrium and brain striatum of Sprague-Dawley rats. Cefoperazone (50 mgkg(-1), i.v.) was then administered via the femoral vein. Blood and brain dialysates were collected and eluted with a mobile phase containing methanol-100 mM monosodium phosphoric acid (30:70, v/v, pH 5.5). The wavelength of the UV detector was set at 254 nm. The detection limit of cefoperazone was 20 ng mL(-1). Isocratic separation of cefoperazone was achieved within 10 min. The intra- and inter-assay accuracy and precision of the analyses were < or =10% in the range of 0.05-10 microg mL(-1). The ratio of the area under the concentration curve of cefoperazone in rat brain and blood was estimated to be about 7-8%. It is concluded that cefoperazone is capable of penetrating the blood-brain barrier. PMID- 11007068 TI - Ventricular function and cardiac hypertrophy after coronary thrombolysis with tissue-type plasminogen activator (t-PA) in dogs with coronary artery thrombi. AB - Subacute prognosis of cardiac function after thrombolysis with a modified tissue type plasminogen activator (t-PA) YM866 was determined in dogs with coronary artery thromboses induced by injection of a thrombin, fibrinogen and autogenous blood mixture. The left ventricular ejection fraction (LVEF) decreased 30 min after occlusion and had not improved 1 week later. Examination after sacrifice revealed myocardial infarction as well as increases in both the left ventricular myocardial area and heart mass. Occluded coronary arteries reperfused by YM866 (0.1 mg kg(-1) i.v.) treatment 30 min after occlusion, by contrast, had improved LVEF and inhibited myocardial infarction development. In addition, the left ventricular myocardial area and heart mass were significantly reduced compared with the vehicle control group 1 week after administration. Although occluded coronary arteries reperfused by YM866 (0.1 mg kg(-1) i.v.) treatment 3 h after occlusion did not show an improvement in the LVEF or inhibition of myocardial infarction development, the left ventricular myocardial area and heart mass decreased significantly compared with the vehicle control group 1 week after administration. In conclusion, early reperfusion by t-PA treatment 30 min after occlusion improved the ventricular function and cardiac hypertrophy, whereas late reperfusion by t-PA treatment 3 h after occlusion did not improve the ventricular function but did inhibit hypertrophy in dogs with coronary artery thrombi. PMID- 11007069 TI - Tiospirone and the reinforcing effects of cocaine in the conditioned place preference paradigm in rats. AB - Tiospirone (TSP) is an atypical antipsychotic drug. It has 5HT-2 antagonistic properties as well as affinity for D2, 5HT-1a, 5HT-6 and sigma receptors. Behavioural studies in our laboratory, which used a 24h free access to food and fluids paradigm, showed a decreased alcohol and increased food intake after twice daily administration of TSP; the maximal effect was obtained at a dose of 0.48 mg kg(-1). This study used the conditioned place preference paradigm to determine the effect of TSP on the reinforcing properties of cocaine. Intraperitoneal administration of 5.0 mg kg(-1) cocaine, but not saline, increased the time rats spent in the drug-paired compartment of a three-compartment shuttle box by 104.9%. Two doses of TSP, 0.143 and 0.48 mgkg(-1), were tested subcutaneously 60 min before saline or cocaine administration during the conditioning phase only. A dose-response effect was observed with a significant reduction in the time rats spent in the cocaine-paired compartment on the drug-free test day produced by the dose of 0.48 mg kg(-1) (an increase of only 38.1% when post-conditioned times were compared with preconditioned times). These findings suggest that TSP reduces the reinforcing properties of cocaine exhibited in the conditioned place preference paradigm. PMID- 11007070 TI - The effects of potassium channel blockers on progesterone-induced suppression of rat portal vein contractility. AB - The suppression of contractility of rat portal vein caused by progesterone appears to be due to the potassium (K+) channel opening effect of this hormone. The identity of the specific K+ channels involved has been investigated using a variety of K+ channel blockers. Incubation with 100 nM iberiotoxin antagonised the progesterone-induced inhibition of spontaneous and 20 mM K+-induced phasic activity of the portal vein such that the contractions resembled those of the non progesterone, non-iberiotoxin control tissues treated with the corresponding solvent vehicles. Incubation with barium chloride (20 and 100 microM), 4 aminopyridine (1 mM), tetraethylammonium chloride (1 mM), glibenclamide (1 microM) or apamin (1 microM) did not, however, have the same antagonistic effect. These results suggest that progesterone's selective suppression of rat portal vein contractility is mediated by the opening of BKCa channels. PMID- 11007071 TI - Effect of Aegle marmelos on biotransformation enzyme systems and protection against free-radical-mediated damage in mice. AB - The effect of hydroalcoholic (80% ethanol, 20% water) extract of leaves of Aegle marmelos was examined on carcinogen-metabolizing phase-I and phase-II enzymes, antioxidant enzymes, glutathione content, lactate dehydrogenase and lipid peroxidation, using two doses of dried extract (50 and 100 mg kg(-1) daily for 14 days), in the liver of mice. The modulatory effect of the extract was also examined on extrahepatic organs (lung, kidney and fore-stomach) for effects on the activity of glutathione S-transferase, DT-diaphorase, superoxide dismutase and catalase. Extract treatment significantly increased the basal levels of acid soluble sulphydryl (-SH) content, cytochrome P450, NADPH-cytochrome P450 reductase, cytochrome b5, NADH-cytochrome b5 reductase, glutathione S transferase, DT-diaphorase, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase in the liver. Aegle acted as a bifunctional inducer since it induced both phase-I and phase-II enzyme systems. Both doses significantly decreased the activity of lactate dehydrogenase and formation of malondialdehyde in liver, suggesting a role in cytoprotection as well as protection against pro-oxidant-induced membrane damage. Butylated hydroxyanisole (positive control) induced almost all the antioxidative parameters measured in this study. The extract was effective in inducing glutathione S-transferase, DT diaphorase, superoxide dismutase and catalase in lung, glutathione S-transferase, DT-diaphorase and superoxide dismutase in fore-stomach, and DT-diaphorase and superoxide dismutase in lung. These significant changes in the levels of drug metabolizing enzymes and antioxidative profiles are strongly indicative of the chemopreventive potential of this plant, especially against chemical carcinogenesis. PMID- 11007072 TI - The effect of acute ethanol exposure on the chronotropic and inotropic function of the rat right atrium. AB - Consumption of ethanol (CH2CH3OH), both acutely and chronically, is known to affect cardiac function and may alter the autonomic control of the heart. This study investigated the effects of two modes of acute exposure to ethanol on the chronotropy and inotropy of the rat right atrium with emphasis on alterations in the adrenergic responses. Atria from rats infused with an anesthetizing level of ethanol for 21 h showed a tendency for a greater increase of the unstimulated beating rate with isoproterenol (ISO), while both unstimulated inotropy and the inotropic response to ISO were significantly decreased compared with the control. Right atria in the presence of ethanol in-vitro demonstrated decreased basal active tension development and decreased inotropic responses to ISO. No alteration of the chronotropic response to ISO was evident with any concentration of ethanol. These results demonstrate both an immediate as well as a persistent effect of ethanol on right atrial chronotropy and inotropy. Alterations in the G stimulatory subunit of the adenylate cyclase system and alterations in myofilament binding of Ca2+ are consistent with these observed ethanol effects. PMID- 11007073 TI - Uptake and dispersion of metformin in the isolated perfused rat liver. AB - Although metformin is a widely used oral antihyperglycaemic, the exact mechanisms of its cellular uptake and action remain obscure. In this study the hepatic extraction and disposition kinetics of metformin were investigated by use of an isolated in-situ rat liver preparation. The liver was perfused in single-pass mode with protein-free Krebs bicarbonate medium at a flow rate of 20 mL min(-1). During constant infusion with 1 mg L(-1) metformin hydrochloride the hepatic uptake of metformin approached equilibrium within 10 min. The steady-state availability, F, determined from the ratio of outflow concentration to input concentration, was 0.99+/-0.02 (mean +/- s.d., n = 4). The outflow profile of metformin resulting from a bolus injection of 25 microg into the portal vein, had a sharp peak then a slower declining terminal phase. The mean transit time (MTT; 49.5+/-14.5, n = 6) and normalized variance (CV2; 4.13+/-0.05) of the hepatic transit times of metformin were estimated by numerical integration from the statistical moments of the outflow data. The volume of distribution of metformin in the liver (1.58+/-0.28 mL (g liver)(-1)) was estimated from its MTT. The volume of distribution is greater than the water space of liver, indicating that metformin enters the hepatic aqueous space and becomes distributed among cellular components. The magnitude of CV2 for metformin is greater than for the vascular marker sucrose, suggesting that distribution of metformin into hepatic tissue is not instantaneous. In conclusion, hepatic uptake of metformin is rate-limited by a permeability barrier. Although metformin is accumulated in the liver, the organ does not extract it. PMID- 11007074 TI - Does dihydrohonokiol, a potent anxiolytic compound, result in the development of benzodiazepine-like side effects? AB - The aims of this study were to assess whether dihydrohonokiol, 3'-(2-propenyl)-5 propyl-(1,1'-biphenyl)-2,4'-diol (DHH-B), a potent anxiolytic compound, developed benzodiazepine-like side effects. A 1 mg kg(-1) dose of diazepam, almost equivalent to the minimum dose for the anxiolytic effect, disrupted the traction performance, potentiated hexobarbital-induced sleeping and impaired learning and memory performance. DHH-B, even at a dose of 1 mg kg(-1) (i.e. five times higher than the minimum dose for significant anxiolytic effect) neither developed diazepam-like side effects nor enhanced the side effects of diazepam. Rather, the potentiation by diazepam of hexobarbital-induced sleeping was reduced by 1 mg kg( 1) DHH-B. Furthermore, mice treated with 10 daily administrations of 1 and 5 mg kg(-1) diazepam, but not 0.2-5 mg kg(-1) DHH-B, showed precipitated withdrawal symptoms characterized by hyper-reactivity, tremor and tail-flick reaction when they were challenged with flumazenil (10 mg kg(-1) i.p.). These results suggest that, unlike the benzodiazepine anxiolytic diazepam, DHH-B is less likely to induce motor dysfunction, central depression, amnesia or physical dependence at the effective dose required for the anxiolytic effect. PMID- 11007076 TI - Radish extract stimulates motility of the intestine via the muscarinic receptors. AB - The effects of radish (Brassica oleraceae, Cruciferae) on gastrointestinal motility were examined using rat intestinal segments with myenteric plexus in vitro and measuring the intestinal transit of charcoal in-vivo. Radish extract (10 microg mL(-1) to 2 mg mL(-1)) caused a dose-dependent increase in contractions of the duodenum, jejunum and ileum, and 1 mg mL(-1) was the maximum effective dose. The largest contraction by the extract was found in ileal segments. The extract-induced (0.5 mg mL(-1)) ileal contraction was remarkably inhibited by pretreatment of segments with atropine (10(-7) M) for 10 min, but not by hexamethonium (0.5 mM). Moreover, antagonists of the muscarinic receptor reduced the radish-induced ileal contraction by a different ratio. The rank order of inhibitory effects was 4-diphenylacetoxy-N-methyl-(2-chloroethyl)-piperidine methiodide (90.5% of control) > tropicamide (67.4%) > pirenzepine (42.8%) > methoctramine (16.7%). Oral administration of radish extract (300-500 mg kg(-1) body weight) to mice remarkably improved the intestinal transit of charcoal, and this was significantly attenuated by co-administration of atropine (50 mg kg( 1)). Taken together, these results suggest that radish extract stimulates gastrointestinal motility through activation of muscarinic pathways. PMID- 11007075 TI - Interaction of drugs and Chinese herbs: pharmacokinetic changes of tolbutamide and diazepam caused by extract of Angelica dahurica. AB - The inhibitory effects of Angelica dahurica root extract on rat liver microsomal cytochrome P450 and drug-drug interactions were studied. The 2alpha- and 16alpha hydroxylase activity of testosterone were most strongly inhibited, with 17.2% and 28-5% of their activity remaining, respectively, after oral administration of A. dahurica extract at a 1 g kg(-1) dose. 6beta-Hydroxylase activity was also inhibited, with 70% of its activity remaining, under the same conditions. In addition, treatment with the extract inhibited the metabolism of tolbutamide, nifedipine and bufuralol. These results showed that the extract inhibited the various isoforms of cytochrome P450 such as CYP2C, CYP3A and CYP2D1. The A. dahurica extract delayed elimination of tolbutamide after intravenous administration at a 10 mg kg(-1) dose to rats. Thus, the extract altered the liver intrinsic clearance. It had little effect, however, on the pharmacokinetic parameters of diazepam after intravenous administration at 10 mg kg(-1). Since diazepam showed high clearance, it underwent hepatic blood flow rate-limited metabolism. Therefore, the change of intrinsic clearance had little effect on hepatic clearance. However, the Cmax value after oral administration of diazepam with extract treatment was four times that with non-treatment. It was suggested that the first-pass effect was changed markedly by the extract. High-dose (1 g kg(-1)), but not low dose (0.3 g kg(-1)), administration of A. dahurica extract increased significantly the duration of rotarod disruption following intravenous administration of diazepam at 5 mg kg(-1). It was concluded that administration of A. dahurica extract has the potential to interfere with the metabolism, by liver cytochrome P450, of other drugs. PMID- 11007077 TI - Induction of apoptosis in human leukaemic cell lines K562, HL60 and U937 by diethylhexylphthalate isolated from Aloe vera Linne. AB - We investigated the effect of diethylhexylphthalate (DEHP) from Aloe vera Linne on the apoptosis of human leukaemic cell lines K562, HL60 and U937 to examine its pharmacological activity. At a level of 10 microg mL(-1) DEHP a significant anti leukaemic effect was observed for all three cell lines, as measured by clonogenic assay. After treatment with 10 microg mL(-1) DEHP for 4 h, agarose gel electrophoresis and flow cytometric analysis confirmed the occurrence of apoptosis. These results indicate that DEHP isolated from Aloe vera Linne has a potent antileukaemic effect, and thus represents a new type of pharmacological activity with respect to human leukaemic cells. PMID- 11007079 TI - Prevention of accidental childhood strangulation. A clinical study. AB - Accidental strangulation is a preventable problem, and there is limited scientific understanding of its mechanism in children. If the amount of external pressure that occludes the airway can be determined, design changes may be made to allow for production of household objects that would break apart at safe pressure levels. A force gauge was applied to the suprahyoid region in 90 children under standardized anesthesia. Three blinded observers performed the study. The anesthesiologist maintained the airway and used a stethoscope to auscultate for breath sounds and monitor the CO2 curves to evaluate obstruction. The recorder noted the numbers from the gauge. A single observer applied the force gauge. Age was the most significant variable in occluding the airway. Obstruction appears to occur at the level of the larynx. Increased knowledge regarding the external pressure required for airway occlusion would allow for the design and manufacture of products with a reduced potential for accidental strangulation. PMID- 11007078 TI - Experience with percutaneous dilational tracheostomy. AB - Percutaneous dilational tracheostomy (PDT) has gained popularity among critical care specialists in the past 10 years. The initial studies in our specialty resulted in essentially banning the procedure as a dangerous substitute for standard operative tracheostomy. Despite this action, more than 1,100 cases of percutaneous tracheostomy have been reported with details on complications. We reviewed all published data and studied 311 patients of our own. A prospective study was performed in 3 groups of patients: 1) 50 patients scheduled for PDT performed in the operating room by a head and neck surgeon (group 1); 2) 50 patients who underwent standard operative tracheostomy performed by the same surgeon (group 2); and 3) 211 patients who underwent bedside PDT by critical care physicians (group 3). The intraoperative complication rates were 0% in group 1, 2% in group 2, and 4% in group 3; the postoperative complication rates were 13%, 4%, and 12%, respectively. There were 2 deaths in group 3, and none in groups 1 or 2. The statistically significant differences among the groups were the superiority of group I over group 3 in intraoperative complications, as well as the lower postoperative complication rate of the standard tracheostomy group. These results show that PDT can be performed with acceptable morbidity rates in relation to published complication rates of standard tracheostomy, but it has no advantage over standard tracheostomy with respect to postoperative morbidity. When they are performed by a head and neck surgeon, the morbidity associated with both standard and percutaneous tracheostomies can be reduced. PMID- 11007080 TI - Dilatational versus standard tracheostomy: a meta-analysis. AB - The advent of percutaneous dilatational tracheostomy (PDT) was initially viewed by otolaryngologists with great skepticism. The purpose of this study was to compare the complications of PDT with those of standard tracheostomy (ST) by a meta-analysis of randomized studies. We found that ST had a fivefold higher rate of complications than did PDT, and these complications were often more severe. We conclude that PDT is a safer procedure for elective tracheostomy in carefully selected patients, ie, those with normal-sized necks. PMID- 11007081 TI - Perioperative airway complications following pharyngeal flap palatoplasty. AB - This study was performed to determine the incidence and types of perioperative airway complications after pharyngeal flap palatoplasty. We conducted a retrospective chart review of 88 patients who underwent correction of velopharyngeal insufficiency between April 30, 1983, and April 30, 1997, in a tertiary care hospital. Some degree of airway obstruction developed in 7 patients. One child developed laryngobronchospasm and required immediate endotracheal intubation. He was successfully extubated without sequelae. Another patient developed severe obstructive sleep apnea and required flap revision. A third patient was found asystolic and apneic. She was immediately intubated; however, she subsequently died. Two patients aspirated blood, presumably resulting in pneumonia. They were managed with parenteral antibiotics. Another child developed worsening sleep apnea and required flap revision. One patient developed nasal obstruction that resolved with time. Airway compromise in patients who undergo pharyngeal flap palatoplasty can be a potentially fatal complication. Careful surveillance should be maintained over patients with underlying neurologic, craniofacial, or cardiopulmonary disorders. PMID- 11007082 TI - Tracheobronchial extension of recurrent respiratory papillomatosis. AB - Endobronchial and pulmonary dissemination reportedly occurs in 5% of patients with laryngeal papillomatosis. It is more frequently observed in the juvenile form, and carries significant implications for treatment. In this study, we review our experiences with pulmonary and endobronchial extension of laryngeal papillomatosis. The records of the 52 patients with laryngeal papillomas treated at our institution since 1980 were reviewed. Twenty-nine percent of the patients (N = 15) developed tracheobronchial extension; 7% (N = 4) demonstrated pulmonary involvement. The birth history, race, sex, age at onset of symptoms, presenting symptoms, site and type of papillomas, surgical procedures, other methods of treatment, complications, and mortality data were analyzed. Eighty percent of the patients with tracheobronchial involvement were born to mothers with a positive history of vaginal condylomas. Pulmonary changes in 4 patients were observed on either chest radiographs or computed tomography and were manifested as multiple parenchymal nodules in 3 of the 4 patients. Pneumatoceles, cavitary empyema, and multiple recurrent pneumonias were the predominant complications of pulmonary involvement. Of the 15 patients with tracheobronchial extension, 80% (N = 12) required tracheotomy before their presentation to us. The number of multiple surgical procedures performed to remove papillomas ranged from 7 procedures in 1 patient with a solitary tracheobronchial lesion to more than 100 procedures in 2 patients with diffuse bronchopulmonary papillomatosis. As our study shows, tracheobronchial involvement continues to complicate surgical treatment in this challenging disease process. PMID- 11007083 TI - Outcomes assessment following treatment of spasmodic dysphonia with botulinum toxin. AB - Spasmodic dysphonia (SD), a disabling focal dystonia involving the laryngeal musculature, is most commonly treated by the intramuscular injection of botulinum toxin (BTX). Although the treatment is well tolerated and generally produces clinical voice improvement, it has never been statistically shown to alter the patient's perception of voice quality or general health. Declining resources for medical care mandate that treatment outcomes be documented. A prospective analysis of the effects of BTX on the patient's perception of voice and general health was undertaken. The Voice Handicap Index (VHI) and Short Form 36 (SF-36) surveys were administered to patients before treatment and 1 month after. Pretreatment and posttreatment scores were analyzed with a Student's t-test. On the VHI, improvements in the patients' perception of their functional, physical, and emotional voice handicap reached statistical significance (p < or = .0005). On the SF-36, patients had statistically significant improvements in mental health (p < or = .03) and social functioning (p < or = .04). Treatment of SD with BTX significantly lessened the patients' perception of dysphonia. In addition, it improved their social functioning and their perception of their mental health. These outcome measures justify the continued treatment of SD with BTX. PMID- 11007084 TI - Acoustic, aerodynamic, and videostroboscopic features of bilateral vocal fold lesions. AB - Successful treatment of bilateral vocal fold lesions depends on the accuracy of the diagnosis. For example, the preferred treatment for vocal fold nodules is voice therapy: in contrast. treatment for a unilateral vocal fold lesion with a contralateral reactive vocal fold lesion (UVFL/RL) usually involves phonosurgery and voice therapy. Differentiation between vocal fold nodules and a UVFL/ RL is often challenging. The purpose of this study was to facilitate diagnostic accuracy and improve treatment for patients with bilateral vocal fold lesions by attempting to identify distinct features of patients with either vocal fold nodules or a UVFL/RL with acoustic, aerodynamic, stroboscopic, and patient self perception measures. The objective voice analysis, Voice Handicap Index, and laryngovideostroboscopic examinations of 85 patients with bilateral vocal fold lesions were reviewed. The results indicated that the patients with a UVFL/RL presented a diagnostic profile that was significantly different from that of patients with vocal fold nodules. Statistically significant differences were found for 1) symmetry of vocal fold vibration, 2) amplitude perturbations, 3) estimated subglottic pressure, and 4) Voice Handicap Index. These results suggest that a composite assessment of acoustic, aerodynamic, and videostroboscopic phonatory features facilitates differentiation between patients with vocal fold nodules and those with a UVFL/RL. The improved diagnostic accuracy afforded by multiparametric assessment provides a comprehensive framework for the treatment of these two distinct vocal fold disorders. PMID- 11007085 TI - Photography of severe laryngeal obstruction. AB - Thirty-five-millimeter photography using rigid rod lens telescopes at direct laryngoscopy is the most versatile and reliable method of laryngeal documentation. Photography of severe laryngeal obstruction, whether in pediatric or adult patients, mandates a method of anesthesia chosen for maximum patient safety, a laryngoscope selected for optimal exposure, confidence in controlling the airway, and a technique that does not jeopardize the safety of the patient. PMID- 11007086 TI - Critical evaluation of neurolaryngological disorders. AB - Otolaryngological examinations, videostroboscopic image analysis, and laryngeal electromyography were used as a test battery for a critical evaluation in 80 patients. Vocal fold movements were categorized into mobility, restricted mobility, immobility with different positions, and overactive movement. Laryngeal electromyographic examinations were conducted in all patients, and the results were classified into normal, neuropathic, and myopathic patterns. The electromyographic data were integrated with videostroboscopic findings, interpreted with knowledge of biomechanical and electrophysiological mechanisms of the larynx, and correlated clinically with underlying diseases. It is suggested that neurolaryngological procedures are most clinically useful when dictated by a decision-making algorithm. PMID- 11007087 TI - Argon plasma coagulation for inferior turbinate reduction. AB - Surgical reduction of the inferior turbinate remains a widely used therapy for nasal obstruction caused by hyperplastic turbinates. Current methods are costly and time-consuming, require nasal packing, and cause extended postoperative swelling. Argon plasma coagulation (APC) is a new, innovative technique in otorhinolaryngological surgery that is based on high-frequency electrical current. The current is transmitted through ionized argon gas (plasma) contact free to the tissue surface and creates a coagulation and devitalization zone with limited penetration. Forty-five patients with nasal obstruction were treated under local anesthesia for reduction of the inferior turbinates with APC. Seventy six percent of the patients reported an improvement of postoperative swelling within the first week. After 3 months, 86% reported having better nasal airflow than they had before the operation. Macroscopically, after 4 weeks the turbinate was covered by normal mucosa in 51% of the patients. Crust formation was minimal, and no bleeding occurred. Turbinate reduction with APC is an alternative technique with many advantages, and the results are comparable to those of standard techniques. Ninety-five percent of the patients were satisfied with the operation. PMID- 11007088 TI - Postoperative radiotherapy in patients with positive nodes after functional neck dissection. AB - A study was designed to assess the usefulness of postoperative radiotherapy (RT) in patients with surgically treated laryngeal and hypopharyngeal cancer with histologically proven positive neck nodes. Patients underwent operation between 1984 and 1995, with functional neck dissection (FND) being part of the treatment in all cases. The selection criteria included squamous cell carcinoma, negative margins for the primary tumor, and no previous treatment. For evaluation purposes, patients were divided into 2 groups: surgery alone versus surgery with postoperative RT. Eighty-three patients fulfilled the inclusion criteria and entered the study. All but 1 of the patients were men. The mean age was 58 years (range, 35 to 77 years). A multivariate analysis was used to analyze the prognostic parameters selected by univariate analysis, eg, age, alcohol, tumor location, T and N stages, and presence or absence of extracapsular spread and a desmoplastic pattern. Postoperative RT was not selected by univariate analysis as a prognostic factor, but was included in the multivariate analysis in order to assess its impact on survival and recurrence rates. Using the statistical method of multivariate analysis, we could not find evidence of a benefit to survival or local recurrence rates with postoperative RT in this series. Patients younger than 55 years and those with extracapsular spread had a decreased survival rate and a higher neck recurrence rate, irrespective of the treatment method. PMID- 11007089 TI - Facial nerve: vascular-related anatomy at the stylomastoid foramen. AB - We dissected 30 facial nerves in fresh cadavers after arterial casting with red latex to provide specific information about the arterial-related anatomy of the trunk of the facial nerve from the stylomastoid foramen to its bifurcation. We found that a wide anatomic variability does exist. The trunk of the facial nerve was in proximity to the stylomastoid artery, which originated from the posterior auricular artery in 70% of the specimens (21/30), from the occipital artery in 20% (6/30), and directly from the external carotid artery in 10% (3/30). The stylomastoid artery passed medially to the trunk of the facial nerve in 63 of the specimens (19/30) and laterally in 37% (11/30). Among these 11 specimens, 8 were large-caliber stylomastoid arteries. During parotid surgery, the main trunk of the facial nerve may be difficult to identify, because a large-caliber stylomastoid artery can mask it. Therefore, it is important to dissect this artery with caution. PMID- 11007090 TI - Postoperative headache after surgery for vestibular schwannoma. AB - Postoperative headache was studied among 251 patients who underwent operation for vestibular schwannoma. A questionnaire based on the McGill Pain Questionnaire and the Finnish Pain Questionnaire was sent to the patients. Twenty-one expressions describing postoperative headache were extracted with a factor analysis. The pain intensity was expressed on a visual analog scale, and the risk factors for postoperative headache were evaluated. Immediately after the operation, 154 subjects reported headache. Eighty-nine of the patients had had headache before operation, whereas 65 patients experienced headache only after operation. An average of 8.9 years after surgery, 93 patients still reported headache. Headache was a major problem for 27 subjects; 18 of the 27 had suffered from headache before operation. A retrosigmoidal approach, postoperative gait problems, preoperative headache, and small tumor size predicted postoperative problems with headache. When headache is present before operation, it tends to continue after operation, and if headache continues for 1 year, it usually persists without being reduced. PMID- 11007091 TI - Artifactual thickening of the sinus walls on computed tomography: a phantom model and clinical study. AB - Measuring sinus wall thickness on computed tomography may be important for distinguishing between acute and chronic inflammation of the paranasal sinuses or in cases of a suspected neoplasm. The objective of this study was to investigate the effects of opacified and aerated sinuses on the appearance of sinus wall thickness. A phantom model consisting of a skull half-immersed in water was scanned, and various slice thicknesses and different windows were used. The sinus walls of the water-immersed side appeared to be thicker than those of the aerated side. Bone windows did not completely eliminate this partial volume effect. It was concluded that comparison between a fluid- or tissue-filled sinus and an air filled counterpart is not accurate enough for evaluating sinus wall thickness. Bone windows do not completely eliminate the artifactual thickening of the bony wall of a filled sinus. PMID- 11007092 TI - Management of large Mohs defects. AB - This study was performed to determine the appropriateness and relative merits of closing Mohs defects of the head and neck at the time of surgery or after surgery, in contrast with allowing defects to heal spontaneously. For 185 patients who had large Mohs wounds of the head and neck (scalp, nose, cheek, forehead, lip, chin, ear, or neck) that healed spontaneously and who underwent postoperative reconstruction as required, we recorded wound location, size, and depth. We then evaluated the cosmesis and functional result periodically and > or = 6 months after operation by chart review, interview, or both. We found that large wounds of the scalp, neck, and ear (except through-and-through defects) often heal spontaneously with acceptable cosmesis. Large defects of the nose usually require prompt reconstruction. Large defects of the central cheek, lip, and chin usually heal spontaneously with poor cosmesis. However, we often reserve surgery for an unacceptable cosmetic result, because repairing a small scar is often less complex than reconstructing a large Mohs defect. We conclude that spontaneous healing of selected Mohs wounds of the head and neck can result in satisfactory cosmesis and function, thus obviating or minimizing the need for complex surgical repair. Surgeons can select management of Mohs wounds effectively and efficiently by accurately predicting the final cosmetic and functional result of spontaneous healing, thus reserving surgery for unfavorable cosmetic or functional results. PMID- 11007093 TI - Treatment of nasal polyposis in Byzantine times. AB - The goal of this study was to describe the therapeutic methods and surgical techniques used during Byzantine times (AD 324-1453) for a disease that has occupied physicians since antiquity: nasal polyps. The original Greek-language texts of the Byzantine medical writers, most of which were published after the 17th century, were studied in order to identify the early knowledge of the definition, symptoms, conservative treatments, and surgical intervention in cases of this disease. A considerable number of conservative treatments, etiologic and local (with inunctions or blowing of caustic substances), with evident influence from Roman medicine, were identified even in the early Byzantine medical texts (4th century). Further, some surgical techniques were described that seem to constitute evolution of the Hippocratic tradition. From the study of the original texts of Byzantine medical writers, their interest in the rhinological diseases is evident; in the case of nasal polyps, new techniques were mentioned. The first meticulous intranasal surgical removal of polyps was described. These techniques, obviously developed during the Hellenistic period, initially influenced European medicine and later the rest of the world. PMID- 11007094 TI - Meningitis due to acute otitis media and arachnoid granulations. PMID- 11007095 TI - Malignant triton tumor of the ethmoid sinus and nasal cavity. PMID- 11007096 TI - Delayed gastric emptying in human immunodeficiency virus infection: correlation with symptoms, autonomic function, and intestinal motility. AB - Gastric emptying may be delayed in HIV infection. We aimed to characterize the pattern of gastric emptying in HIV seropositive subjects and correlate the findings with symptoms, as well as to identify possible etiological factors. Solid gastric emptying was measured using scintigraphy in 54 HIV seropositive subjects and 12 HIV seronegative controls. Gastrointestinal symptoms were evaluated using a standardized numerical score, and autonomic function was assessed using spectral analysis of heart rate variability. Fasting and postprandial duodenojejunal activity was recorded using strain gauge manometry catheters. Gastric emptying rate, but not lag phase, was significantly delayed in HIV-infected subjects, particularly those with enteric infections and more advanced disease. Delayed gastric emptying did not correlate with symptoms, autonomic dysfunction, or small intestinal motility. In conclusion, abnormalities found in autonomic function and gastric emptying in HIV infection are multifactorial in nature. The contribution of upper gastrointestinal motor dysfunction to gastric symptoms in such individuals is unclear. PMID- 11007097 TI - Histomorphometry and strain distribution in pig duodenum with reference to zero stress state. AB - The morphometry at no-load and zero-stress states and residual circumferential strains were determined along the pig duodenum in vitro in seven pigs. The no load state was obtained by cutting eleven 2-mm-wide rings at 10% intervals along the duodenum. The zero-stress state was obtained by cutting the rings radially. The zero-stress state provides a standard morphological state to describe tissue since internal and external forces do not affect the tissue. The morphometric measures were obtained from digitized images, and the layer thicknesses were measured from histological sections. The mucosal and serosal circumferences, the wall thickness, and the wall thickness-to-mucosal radius ratio were largest in the proximal end of the duodenum (f > 1.9, P < 0.05). The thickness of the submucosal stratum compactum layer and the opening angle increased in distal direction (f = 2.3, P < 0.05 and = 6.5, P < 0.001). The residual strain at the mucosal surface was negative, indicating that the mucosa-submucosa layers of duodenum in no-load state are in compression. Distension experiments showed that the residual strain makes the stress distribution through the wall more uniform in the pressurized state. In conclusion, the large circumferential residual strains must be taken into account in the study of physiological problems, in which the stresses and strains are important, eg, the bolus transport function. PMID- 11007098 TI - Determinants of occurrence and volume of transpyloric flow during gastric emptying of liquids in dogs: importance of vagal input. AB - The precise factors and their relative contributions that lead to individual flow pulses across the pylorus during liquid gastric emptying remain unclear. Our objective was to determine the factors leading to individual flow pulses, their relative contributions and the role of the vagus nerve in their modulation. Proximal gastric tone had a strong positive correlation with the volume of the corresponding transpyloric flow pulse whereas pyloric tone had an inverse correlation. Antral contractions were associated with the presence but not the volume of the pulse. Acute vagal blockade retarded emptying via loss of proximal gastric tone and increased outflow resistance and loss of propagating antral pressure waves. In conclusion, the major determinants of the volume of pulsatile transpyloric flow are proximal gastric and pyloric tone. The vagus nerve plays a key role in regulating both proximal gastric and pyloric tone as well as moderating propagating antral contractions. PMID- 11007099 TI - Alterations of GTP-binding proteins (Gsalpha and Gq/11alpha) in gastric smooth muscle cells from streptozotocin-induced and WBN/Kob diabetic rats. AB - We investigated possible impairment of the signal transduction system in gastric myocytes of streptozotocin-induced diabetic (STZ) and spontaneous diabetic WBN/Kob (WBN/Kob) rats. Gastric motility 10 weeks after the onset of diabetes mellitus was significantly reduced in both diabetic rats compared with control, and the decreased motility was not recovered by the administration of insulin to maintain normal blood glucose levels. There was no significant difference between both types of diabetic rats and control rats in total number of [3H]quinuclidinyl benzilate ([3H]QNB) binding sites (Bmmax: 545-587 fmol/mg protein) on gastric smooth muscle cell membranes or in the affinity of [3H]QNB for the binding sites (Kd: 0.06-0.07 nM). Immunoblot analysis using polyclonal anti-G-protein antibodies indicated increased expression of Gsalpha in gastric smooth muscle cell membranes, but no significant change in Gialpha or Gq/11alpha expression in STZ rats, and decreased expression of Gq/11alpha with no significant change in Gsalpha and Gialpha in WBN/Kob rats. The cAMP production in gastric smooth muscle cell membranes was augmented in the absence and presence of 100 microM isoproterenol, and 100 microM forskolin in STZ rats, whereas no significant change of cAMP production was observed in WBN/Kob rats irrespective of the presence of the stimulants. These findings suggest that long-standing diabetes may induce alterations in signal transduction at downstream receptors in gastric myocytes, resulting in the impairment of gastric motility, although the mechanism of reduced contractile activity may differ between STZ and WBN/Kob rats. PMID- 11007100 TI - Distal gastrectomy and Roux-Y limb in the rat: plasma motilin, pancreatic polypeptide concentrations, and duodenojejunal motility. AB - After gastrectomy, Roux-Y limb reconstruction leads to duodenojejunal motor disturbances. Because motilin and pancreatic polypeptide (PP) play a role in the regulation of digestive motility, their plasma concentrations were determined in rats after Roux-Y gastrectomy. Three months after a distal Roux-Y gastrectomy, coupling of electromyographic recordings and jugular samples were used to perform motilin and PP radioimmunoassays during and between activity fronts (AFs) occurring in the limb and in the duodenojejunum, 20, 40, and 60 min after intragastric instillation of a standard meal (5 ml Realmentyl). Animals that underwent a simple laparotomy, animals having isolated jejunal transection, and animals with Billroth I gastrectomy (BI group) served as control groups. After Roux-Y gastrectomy, the number of AFs in the limb (P < 0.01) and in the duodenum (P < 0.001) was reduced compared to laparotomized rats and the BI group, but did not differ from the number in the Tr group. In the limb, AFs were incompletely propagated or were retrograde in 9 and 3 of 20 animals, respectively. After Roux Y gastrectomy, motilin concentrations occurred at the same time as each duodenal AF, and as in controls, and were independent from AFs in the limb. Plasma motilin concentrations were higher after Roux-Y reconstruction than in control groups (P < 0.03), and PP level concentrations were not different. After the meal, the interruption of AFs was shorter in Roux-Y reconstruction than in laparotomized and transected animals (P < 0.05) and than in BI group, with no significant difference in the latter. In all groups, plasma motilin and PP concentrations were decreased (P < 0.05) and increased (P < 0.001), respectively, after the meal compared to the interdigestive period. After Roux-Y gastrectomy, plasma motilin and PP levels were higher (P < 0.05) and lower (P < 0.05), respectively, compared to controls. In conclusion, AFs in the Roux-Y limb were not associated with plasma motilin concentrations, suggesting a lack of influence of motilin on the interdigestive motor status. The decrease in postprandial plasma PP concentrations may play a role in the shorter interruption of AFs after a meal. PMID- 11007101 TI - Association of Zollinger-Ellison syndrome with pancreatitis: report of five cases. AB - In a retrospective analysis, five cases of Zollinger-Ellison syndrome were found in a typical urban inner-city teaching hospital. Chronic alcohol abuse and heavy smoking characterized these patients, and four of them also had pancreatitis, suggesting an association of gastrin-producing tumors and pancreatic inflammation. Ductal obstruction by neuroendocrine tumors has been reported to cause pancreatitis in a few cases. In this analysis, however, a nonobstructive gastrinoma was the surgical diagnosis in three patients, and it was suggested by imaging studies in the two other cases. The potential other pathomechanisms for a dual cause-effect relationship of gastrinoma and pancreatitis are discussed. PMID- 11007102 TI - P-selectin expression and Kupffer cell activation in rat acute pancreatitis. AB - This work studied the activation of hepatic macrophages during acute pancreatitis and the involvement of these cells in the lung inflammatory response. Pancreatitis was induced in Wistar rats by intraductal administration of 5% sodium taurocholate. Three hours after pancreatitis induction, the degree of pulmonary inflammation, TNF-alpha levels, and P-selectin expression were evaluated. The generation of TNF-alpha by Kupffer cells was also measured. Pancreatitis increases the serum concentration of TNF-alpha, neutrophil infiltration, and P-selectin expression in pancreas and lung. In addition, Kupffer cells generate increased levels of TNF-alpha. When Kupffer cells were inhibited, the increase in serum TNF-alpha levels and the infiltration of neutrophils in the lung were prevented, but P-selectin expression remained unmodified. We conclude that pulmonary inflammation induced by acute pancreatitis is mediated by Kupffer cell activation and that pancreatitis induces the expression of P-selectin on pulmonary endothelial cells but this effect is not mediated by Kupffer cells. PMID- 11007103 TI - Chronic hyperlipasemia caused by sarcoidosis. AB - A chronically elevated lipase is a rare biochemical finding and has only previously been described in patients with malignancy and macrolipasemia. We report a case of chronic hyperlipasemia caused by sarcoidosis. The literature on pancreatic sarcoidosis is reviewed and the significance of lipase isoforms is discussed. Sarcoidosis needs to be considered in patients presenting with chronic hyperlipasemia. PMID- 11007104 TI - Efficacy of lactulose in cirrhotic patients with subclinical hepatic encephalopathy. AB - To investigate the role of lactulose in the treatment of cirrhotic patients with subclinical hepatic encephalopathy (SHE), 40 cirrhotic patients, 33 males and 7 females, were included in the study. The diagnosis of SHE was made by quantitative psychometric tests including the number connection test (NCT), figure connection test (FCT) parts A and B, and two performance subtests of Wechsler adult intelligence scale, ie, picture completion (PC) and block design (BD) tests. SHE was diagnosed in 26 (65%) of 40 patients. Of these 26 patients, 14 patients were randomized to treatment group (lactulose 30-60 ml/day for three months, SHE-L) and 12 patients to no treatment group (no lactulose, SHE-NL). Psychometric tests were repeated in all patients in both groups and in six patients with no SHE (group NSHE, N = 14) after three months. The mean scores and number of the abnormal psychometric tests at entry were significantly higher in patients in groups SHE-L and SHE-NL than in patients in group NSHE; however, there was no significant difference between SHE-L and SHE-NL. The mean number of the abnormal psychometric tests decreased in patients in group SHE-L after three months of treatment with lactulose (2.9 +/- 0.9 vs 0.8 +/- 1.2; P = 0.004); however, there was no change in patients in group SHE-NL after three months (3.7 +/- 1.5 vs 3.5 +/- 1.3; P = NS). While SHE improved in 8 of 10 patients in group SHE-L, none of the patients in group SHE-NL improved after three months of follow up (P < 0.001). Two patients in group SHE-NL also developed overt encephalopathy during the study period. We conclude that lactulose treatment in cirrhotic patients with SHE is effective. PMID- 11007105 TI - Acetaminophen hepatoxicity. AB - To determine the influence of psychosocial factors in accidental and deliberate acetaminophen overdose, we reviewed the charts of 207 overdose patients, and 48 met our criteria for acetaminophen toxicity. Two patients died. A psychiatric history was present in 75%, and 25% had a previous or subsequent suicide attempt. A substance abuse history was elicited from 46% and 36% of adolescent teenagers had a teen pregnancy. The mean time to starting N-acetylcysteine was 18.5 hr. Delayed N-acetylcysteine administration led to higher transaminase levels. Alcohol abuse was associated with a longer hospital stay. Mean AST was 8,860 IU/liter in the accidental and 3,013 IU/liter in the suicide groups. We concluded that management of acetaminophen toxicity can be optimized by early identification, obtaining a complete drug screen, starting N-acetylcysteine early or whenever toxic acetaminophen levels or elevated transaminases are identified, and referring patients with acetaminophen toxicity to a liver center. PMID- 11007106 TI - Ursodeoxycholic acid treatment in abdominal sarcoidosis. PMID- 11007107 TI - Hyperplastic liver nodules associated with early-stage primary biliary cirrhosis mimicking hepatocellular carcinoma. PMID- 11007108 TI - Electrochemotherapy for colorectal cancer with commonly used chemotherapeutic agents in a mouse model. AB - We examined here the usefulness of electrochemotherapy against colorectal cancer (CRC) using a mouse model. Electropermeabilization profoundly increased the sensitivity of murine CRC, Colon 26, and MC38 cells to bleomycin (BLM) but not to 5-fluorouracil (5-FU) or to cisplatin (CDDP) in vitro. In vivo experiments revealed that electrochemotherapy with 5-FU, CDDP, or BLM was much more effective against CRC compared with the treatment of the drug alone. Electrochemotherapy with BLM or CDDP exhibited profound antitumor effects on subcutaneously established CRC in mice, and complete tumor regression was observed in five and four of eight animals, respectively. Electrochemotherapy with 5-FU also had an impact on CRC development, and complete cure was observed in one of eight animals. Subsequent analyses revealed that electropermeabilization significantly increased intratumoral amounts of BLM and CDDP but not 5-FU. These results indicate that electrochemotherapy may be a promising treatment modality against CRC. PMID- 11007109 TI - 5-aminosalicyclic acid and olsalazine inhibit tumor growth in a rodent model of colorectal cancer. AB - The ability of 5-aminosalicylic acid and olsalazine to inhibit colonic aberrant crypts and tumors was investigated in 1,2-dimethylhydrazine-treated rats. The effect of these drugs on the rates of tumor apoptosis and proliferation was studied as potential mechanisms for their action. 5-Aminosalicylic acid reduced the number of aberrant crypt foci by over one third, while olsalazine had no effect on this parameter. However, both agents effectively reduced tumor number and load, increased the rate of tumor apoptosis, and reduced the rate of tumor cell proliferation. In conclusion, 5-aminosalicylic acid and olsalazine are both ultimately effective chemopreventive agents in this model; however, only 5 aminosalicylic acid inhibited the formation of aberrant crypt foci. The inhibitory effect of these agents in tumors is related to the inhibition of proliferation and the induction of apoptosis. PMID- 11007110 TI - Palliative decompression of obstructive hilar malignancies utilizing an extrahilar biliary approach. AB - Hilar cancers carry a dismal prognosis. Palliation of obstructive jaundice in patients with hilar cancer can be achieved by either surgical or nonsurgical means. Selection of the appropriate palliative measures is a challenging problem. Segmental bilioenteric anastomosis procedures were performed on 19 patients with hilar cancer. Seventeen of the bypasses were done to the segment III duct, known as the ligamentum teres approach, and two bypasses were to the segment V duct. Five patients, who had already been stented percutaneously or endoscopically, were operated on after the stents were clogged and a duodenal obstruction ensued. There were two postoperative deaths (10.5%) and four postoperative complications (21%). All of the 17 surviving patients experienced improvement in the level of jaundice postoperatively and the levels of serum total and direct bilirubin decreased by 78.9% and 84.2%, respectively. Two patients developed late cholangitis before death and were treated by external biliary drainage; one developed duodenal obstruction and was treated by gastrointestinal anastomosis. The mean length of hospital stay was 15.2 days. Mean survival was 8.2 months and the mean period of well-being was 7.8 months. Median survival was 7 months and median period of well being was 7 months. Three patients are still alive at 8, 8, and 24 months. These data suggest that the ligamentum teres approach offers effective palliation for patients with unresectable hilar cancer. PMID- 11007111 TI - Influence of disease site and activity on peripheral neutrophil function in inflammatory bowel disease. AB - Reactive oxygen species, released by phagocytes, are involved in tissue injury in inflammatory bowel diseases. The aim of our study was to evaluate peripheral neutrophil function in patients with ulcerative colitis (N = 66) and Crohn's disease (N = 62) with respect to disease activity and extent, using chemiluminometry after three stimuli. Twenty-seven healthy subjects were enrolled as controls. Neutrophils from ulcerative colitis and Crohn's disease patients had a significantly higher response than those from controls following phorbol myristate acetate (86.6 +/- 6.5, 173.8 +/- 11.9, 167.5 +/- 12.2 mV, P < 0.0001), formyl-methionyl-leucyl-phenylalanine (39.5 +/- 3.4, 41.3 +/- 2.7, 58.6 +/- 4.7 mV, P < 0.001), and zymosan (142.6 +/- 10.4, 223.7 +/- 8.9, 231.2 +/- 9.5 mV, P < 0.0001) administration. The increased response was observed during both active disease and remission. The highest chemiluminescence values were found in patients with active ulcerative pancolitis and ileal Crohn's disease. The activation of circulating neutrophils may indicate persistent intestinal inflammation or may be triggered by luminal factors even in the absence of symptoms. PMID- 11007112 TI - Sequential group trial to determine gastrointestinal site of absorption and systemic exposure of azathioprine. AB - Azathioprine (AZA) is used in the treatment of patients with refractory inflammatory bowel disease; however, its use is limited because of systemic toxicity associated with long-term use. Ileocecal delivery of AZA might be advantageous if local intestinal therapeutic effects could be provided with decreased systemic side effects. Decreased cecal systemic absorption would allow higher dosages of AZA to be administered. A two-phase study was performed to compare the systemic exposure of AZA and 6-mercaptopurine (6-MP) following administration of AZA into the stomach, jejunum, and cecum and to compare the systemic exposure to AZA and 6-MP following administration of three different dosages of AZA into the cecum. In phase I, six healthy male volunteers received three 50 mg sequential doses of AZA via an oral tube directly placed into the stomach, jejunum, and cecum, respectively. In phase II, six healthy male volunteers received three different dosages (50, 300, 600 mg of AZA) into the cecum. Plasma concentrations of AZA and 6-MP at various times were quantified and area under the plasma concentration-time curve (AUC) and mean residence time (MRT) were determined. No significant differences in the AUC of AZA were seen at the different sites. The AUC of 6-MP following administration of AZA into the jejunum (67.0 +/- 30.1 ng x hr/ml) was higher compared to the stomach (39.9 +/- 38.1 ng/hr/ml) and cecum (29.2 +/- 10.9 ng x hr/ml). Jejunal absorption was 68% higher than absorption from the stomach and 129% higher than that of the cecum. Gastric absorption was 27% higher than that of the cecum. Increased dosages given into the cecum resulted in increased AUCs of AZA and 6-MP. The AUCs of AZA following 50, 300, and 600 mg dosages were 16.9 +/- 7.4, 52.3 +/- 67.2, and 132 +/- 151 ng x hr/ml, respectively, and the AUCs of 6-MP were 22.2 +/- 14.9, 63.4 +/- 50.6, and 104 +/- 115 ng x hr/ml, respectively. Systemic exposure to 6-MP is reduced following administration of AZA into the cecum, most likely secondary to reduced absorption of 6-MP from the colon. Higher dosages of AZA presented to the cecum do result in increased systemic absorption, but may still allow more drug to be administered with less toxicity than the same dose received orally. PMID- 11007113 TI - Rebamipide, an antiulcer drug, prevents DSS-induced colitis formation in rats. AB - This study was conducted to investigate the efficacy of rebamipide against experimental colitis induced by dextran sulfate sodium (DSS) in a rat model of inflammatory bowel disease. Experimental colitis was induced in male Wistar rats by oral administration of 3% DSS solution for one week. The rats were provided with standard diet containing 0.105% rebamipide (160 mg/kg/day) for 1 week. In rats treated with rebamipide, clinical (body weight loss, bloody diarrhea, reduced physical activity, severe anemia, shortened colonic length, and perianal injury) and histopathological (pathological lesion score) findings of DSS colitis were significantly less than in rats with DSS colitis not treated with rebamipide. Rebamipide thus inhibited the induction of colitis. Rebamipide significantly reduced concentrations of both interleukin-1alpha and GRO/CINC-1 (IL-8-like substance) and cell infiltrates in colonic wall, in parallel with decreased activity of myeloperoxidase. It also reduced expression of IL-1 mRNA but did not influence expression of GRO/CINC-1 mRNA. The attenuation of colonic indices of colitis by rebamipide in this rat model suggests that this drug might have beneficial effects in the treatment of human ulcerative colitis. These effects of rebamipide are attributable to its inhibition of inflammatory cytokine mediated granulocyte (neutrophil) infiltration into the colon. PMID- 11007114 TI - Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC 15708 and Lactobacillus acidophilus ATCC 4356. AB - The antioxidative effect of intact cells and intracellular cell-free extracts of intestinal lactic acid bacteria B. longum (ATCC 15708) and L. acidophilus (ATCC 4356) was investigated. Both intact cells and intracellular cell-free extracts of 10(9)cells of B. longum and L. acidophilus demonstrated antioxidative activity, inhibiting linoleic acid peroxidation by 28-48%. This indicated that these two strains demonstrated excellent antioxidative activity. B. longum and L. acidophilus also showed the ability to scavenge alpha,alpha-diphenyl-beta picrylhydrazyl (DPPH) free radical, scavenging 21-52%. The intact cells of these two intestinal bacteria demonstrated a high inhibitory effect on the cytotoxicity of 4-nitroquinoline-N-oxide (4NQO). Cytotoxicity of 4NQO was reduced by L. acidophilus by approximately half and by almost 90% by B. longum. Nevertheless, no inhibition of cytoxicity observed for intracellular cell-free extracts of 10(9) cells of B. longum and L. acidophilus. The effect of B. longum and L. acidophilus on inhibiting plasma lipid peroxidation was also evaluated. The results showed that both intestinal strains were able to protect plasma lipid from oxidation at different degrees. The inhibition rates on plasma lipid peroxidation ranged from 11 to 29% for 10(9) cells of B. longum and L. acidophilus. Generally speaking, B. longum demonstrated better antioxidative ability than L. acidophilus in this study. PMID- 11007115 TI - Short-chain fatty acids induce cytoskeletal and extracellular protein modifications associated with modulation of proliferation on primary culture of rat intestinal smooth muscle cells. AB - Short-chain fatty acids are the main end products of bacterial fermentation of carbohydrates. Their role on the metabolism and biology of colonocytes is now well characterized. However, the functional consequences of their presence on intestinal smooth muscle cells remain poorly studied. We aimed to assess the effect of different short-chain fatty acids on ileal and colonic smooth muscle cells in primary culture and on A7R5 line. Butyrate (above 0.1 mM) inhibited A7R5 cell proliferation, while at low concentration (0.05 to 0.5 mM) butyrate significantly stimulated the proliferation of ileal and colonic myocytes in primary culture. An inhibition was observed at higher concentrations. Collagenous and noncollagenous protein synthesis was stimulated by butyrate. Moreover, butyrate stimulated actin and myosin expression. Thus, butyrate, which is produced by dietary fiber fermentation, may affect intestinal muscles by directly acting at the molecular level on myocytes. PMID- 11007116 TI - Reaggregation of rat dissociated myenteric plexus in extracellular matrix gels. AB - The aim of this study was to investigate the growth behavior of freshly dissociated myenteric plexus in a three-dimensional extracellular matrix (ECM) environment with and without stimulation of glial cell line-derived neurotrophic factor (GDNF). Therefore, cell suspensions of the dissected myenteric plexus of newborn rats were cultured in freshly prepared gels of commercially available mixtures of collagen, laminin, and hepatoglycans as a first step towards mimicking the natural environment of the myenteric plexus. The cultures were kept either in chemically defined serum-free medium alone or supplemented with GDNF. Cultures on polylysinc-coated glass cover slips served as controls. Dissociated myenteric plexus grown on polylysine formed dense clusters of neurons with radially outgrowing nerve fibers, while the neurons cultured in the gel reaggregated to much smaller clusters. These contained, depending on the culture conditions, 2-10 neurons. The morphology of the network that was seen in the gels after a few days in vitro resembled very closely the in situ situation of the submucous plexus and the myenteric plexus in hypoganglionic children. Electron microscope investigations showed a high degree of organization with fiber bundles and vesicle-containing varicosities and growth cones. Independent of the method of culturing, GDNF obviously influenced the growth behavior of the dissociated plexus. The size of the ganglia was larger, and the secondary network denser when GDNF was supplemented. Moreover, the enteric neurons in the gel cultures tended to be larger in size when treated with GDNF. Three-dimensional cultures of dissociated myenteric plexus in an ECM gel might be a valuable tool towards the understanding of the formation of the enteric nervous system during development, especially considering pathological conditions such as Hirschsprung's disease or other dysganglionic diseases. PMID- 11007117 TI - Diagnosis and removal of cecal whipworm infection: case report and review. PMID- 11007118 TI - Acute on chronic intestinal pseudoobstruction responds to neostigmine. PMID- 11007119 TI - Influence of smoking and Helicobacter pylori on gastric phospholipids. AB - Smokers show higher rates of peptic ulcer disease (PUD) than nonsmokers, probably due to detrimental effects on the gastric mucosa. Surface-active phospholipids (SPL) are believed to play a key part in gastric cytoprotection. The aim of this study was to determine the chronic effects of smoking on the gastric SPL and to relate them to H. pylori (Hp) -induced effects. Gastric juice was aspirated in 52 patients, with normal findings at planned upper gastrointestinal endoscopy, and concentrations of seven phospholipid subclasses were analyzed. Concentrations of lysophosphatidylethanolamine (1-PE) were increased (P = 0.006) in smokers compared to nonsmokers in non-Hp-infected samples. Nonsmokers infected with Hp showed increased levels of 1-PE (P = 0.01) and phosphatidylinositol (PI) (P = 0.02) compared to subjects not infected. In human gastric juice PI seems to be the dominating PL subclass, in contrast to the composition in biopsy specimens. We also found both Hp-infected and smoking subjects to have higher concentrations of more polar phospholipid subclasses, ie, 1-PE, making the mucosa more vulnerable to acid attack as the gastric surfactant becomes less hydrophobic. PMID- 11007120 TI - Helicobacter pylori seroconversion in asymptomatic blood donors: a five-year follow-up. AB - Several techniques have been developed to diagnose Helicobacter pylori infection and two noninvasive methods are available: carbon 13-urea breath test (UBT) and serology. Measurement of IgG serum antibodies by enzyme-linked immunosorbent assay (ELISA) is a reliable and inexpensive method for detection of infection. The aim of this study was to assess the seroconversion by different techniques after five to eight years. In 1990, 588 of 1,010 asymptomatic donors were found to be seronegative by ELISA, based on an H. pylori whole-cell suspension lysate (sensitivity and specificity: 92% and 97%). In 1995 serum samples from 418 of 588 seronegative donors were collected and retested using the same antigen. 411 of 418 samples were frankly negative, and 7 donors were found to be seroconverted. This group of seven sera represents the object of the study. They were retested by ELISA and western blotting using a different antigen (NCTC). To standardize our techniques, sera from 43 H. pylori positive and 47 H. pylori negative patients according to culture, histology, urease test, and UBT were used. The cutoff for ELISA-NCTC was 0.53 AI (absorbance index) (mean value + 2 SD), and for western blotting was negativity for CagA or <10 bands (sensitivity and specificity: 95% and 96%; 98% and 81% for ELISA and western blotting respectively). According to the results obtained in 1990 and 1995, seven donors were found to be seroconverted by ELISA using sonicated antigen; in five the seroconversion was confirmed by ELISA using NCTC antigen and in two there was concordance with WB. Four of the seven donors were contacted and asked to undergo UBT and a further serum sample was drawn to be reassessed in 1998. A seroconversion was found in all four donors by ELISA, while WB and UBT confirmed the seroconversion in only three of four donors. In conclusion the in-house ELISA used performed well compared to other theoretically better serologic assays and confirmed the low seroconversion rate for H. pylori infection in adult populations living in developed countries. PMID- 11007121 TI - Use of simultaneous high-resolution endoluminal sonography (HRES) and manometry to characterize high pressure zone of distal esophagus. AB - The purpose of this investigation was to separate the high pressure zone (HPZ) of the distal esophagus into its two components, the intrinsic lower esophageal sphincter (LES) and the extrinsic crural diaphragm (CD), using simultaneous esophageal manometry and high-resolution endoluminal sonography. Five normal subjects were studied during end inspiration using a dual manometry/ultrasound catheter. The HPZ in the distal esophagus was characterized ultrasonographically as the CD distally and as an overlap of CD and LES proximally. In four of five volunteers, the initial distal rise in pressure at the HPZ corresponded to imaging of CD rather than imaging of the LES. In all subjects, peak pressure corresponded to an overlap of CD and LES. In conclusion, it is possible to divide the HPZ into its two components, the LES and CD using simultaneous high resolution endoluminal sonography and esophageal manometry. During end inspiration, the CD contributes to the initial distal rise in pressure at the HPZ. Peak pressure of the HPZ corresponds to an overlap of the LES with the CD. PMID- 11007122 TI - Method to obtain endoscopic esophageal samples for primary cell culture: focus on infectious contamination. AB - Cell culture techniques hold great importance for the development of molecular biology. However, when used to study oncology, most of the samples come from surgical specimens. Endoscopy is a interesting alternative to get samples for culture. We studied a protocol to allow the control of infectious contamination potentially related to endoscopy, which could preclude it as a method to obtain cells for culture. Esophageal biopsies from 30 patients were taken through upper gastrointestinal endoscopy, using a previously flamed forceps, and were cultivated with and without amphotericin. Our results showed contamination in 3.3% of the wells without the antifungal and in 0.8% of those with it. Regarding the 30 cases studied, the described protocol was able to provide samples free of contamination in all of them. PMID- 11007123 TI - Barrett's esophagus. AB - Barrett's esophagus occurs more frequently than previously anticipated. Detection of Barrett's esophagus is by endoscopic biopsy in which normal squamous epithelium of the esophagus is replaced by a specialized columnar epithelium of any length. Patients with more than five years of gastroesophageal reflux symptoms, particularly those 50 years of age or older, should have upper endoscopy to detect Barrett's esophagus. With recognition of Barrett's esophagus as a premalignant lesion, the crucial issue is surveillance for detection of dysplasia. Although the natural history of dysplasia is incompletely defined, it is clear that patients with dysplasia have a higher risk for adenocarcinoma than those without dysplasia. Dysplasia is not the ideal marker for selecting patients at high risk for adenocarcinoma, however; recent studies have shown that p53 protein accumulation appears to be earlier and more specific/sensitive marker of malignant potential in Barrett's esophagus. Management of Barrett's esophagus often involves a multidisciplinary evaluation and its current status is reviewed. PMID- 11007124 TI - Geranylgeranylacetone protects cultured guinea pig gastric mucosal cells from indomethacin. AB - One of the major side effects of nonsteroidal antiinflammatory drugs, such as indomethacin, is gastropathy. The purpose of this study was to search for a therapeutic drug to prevent this side effect in vitro. We found that geranylgeranylacetone, a unique antiulcer drug with a heat-shock protein-inducing ability, protected cultured guinea pig gastric mucosal cells from cell damage caused by indomethacin. This cytoprotective effect of geranylgeranylacetone required concentrations of more than 10(-6) M and incubation periods of longer than 2 hr. Pretreatment of cells with an inhibitor of protein synthesis completely abolished the cytoprotective effect of geranylgeranylacetone, suggesting that some proteins induced by the drug are responsible for the cytoprotection. Since pretreatment of cells with low concentrations of ethanol, which also induced the heat-shock proteins, made cells resistant to indomethacin, heat-shock proteins are candidates for the proteins that are involved in the cytoprotective effect of geranylgeranylacetone against indomethacin. PMID- 11007125 TI - Evaluation of individual symptoms cannot predict presence of gastric hypersensitivity in functional dyspepsia. AB - Recently, the concept of gastric hypersensitivity was introduced as an important factor in the pathophysiology of functional dyspepsia (FD), but it is unclear which symptoms can predict the presence of gastric hypersensitivity. Therefore, we evaluated the relationship between common symptoms of FD and various parameters measured by gastric barostat in FD patients. Gastric barostat tests were performed in 64 FD patients and 20 healthy control subjects without gastrointestinal symptoms. Individual symptoms such as early satiety, postprandial fullness, sense of delayed emptying, nausea, vomiting, and epigastric soreness were collected and graded as mild to severe. Basal tone, gastric compliance, and postprandial receptive relaxation were similar in controls and patients, the threshold of abdominal discomfort was lower in FD patients than in controls (8.9 +/- 3.6 mm Hg and 14.5 +/- 3.7 mm Hg, respectively, P < 0.05). However, there were no significant differences in the threshold of abdominal discomfort according to the severity of individual symptoms. In conclusion, a simple evaluation of individual symptoms could not predict the presence of gastric hypersensitivity. PMID- 11007126 TI - Cyclic nucleotide-gated channels in non-sensory organs. AB - Cyclic nucleotide-gated channels represent a class of ion channels activated directly by the binding of either cyclic-GMP or cyclic-AMP. They carry both mono and divalent cations, but select calcium over sodium. In the majority of the cases studied, binding of cyclic nucleotides to the channel results in the opening of the channel and the influx of calcium. As a consequence, cytosolic free calcium levels increase leading to the modifications of calcium-dependent processes. This represents and important link in the chain of events leading to the physiological response. Cyclic nucleotide-gated channels were discovered in sensory cell types, in the retina, and in olfactory cells, and were extensively studied in those cells. However, it is becoming increasingly evident that such channels are present not only in sensory systems, but in most, if not all, cell types where cyclic nucleotides play a role in signal transduction. A hypothesis is presented here which attributes physiological importance to these channels in non-sensory organs. Four examples of such channels in non-sensory cells are discussed in detail: those in the liver, in the heart, in the brain, and in the testis with the emphasis on the possible physiological roles that these channels might have in these organs. PMID- 11007127 TI - Histamine evoked sustained elevations of cytosolic Ca2+ in bovine adrenal chromaffin cells independently of Ca2+ entry. AB - Whole-cell patch-clamp experiments and optical measurements with the Ca2+ fluorescent dye fura-2 were performed to examine histamine induced cytosolic Ca2+ changes in bovine adrenal chromaffin cells. The purpose of this study was to find out whether the sustained plateau phase, which followed the rapid transient increase, was due to Ca2+ influx. The extracellular Ca2+ dependence appeared to be minor, because substitution of Ca2+ with EGTA or BAPTA did not cause obvious changes in the biphasic Ca2+ response. Application of histamine in a Mn2+ containing external solution did not quench the fura-2 signal. It was neither possible to detect a histamine induced depolarisation, nor a Ca2+ permeable current. Changing the driving force for Ca2+ during the plateau phase did not result in a correlating fura-2 signal. Metal ions like Cd2+, La3+ and Co2+ which are known to block Ca2+ influx were unable to abolish the typical histamine induced Ca2+ response. These results suggest that primarily intracellular Ca2+ was responsible for generating the characteristic biphasic Ca2+ response due to histamine in bovine adrenal chromaffin cells. PMID- 11007128 TI - Spatial and temporal aspects of ACh-induced [Ca2+]i oscillations in porcine tracheal smooth muscle. AB - This study evaluated the relationship between regional elevation in intracellular calcium concentration ([Ca2+]i) induced by acetylcholine (ACh) and the global cellular responses in porcine tracheal smooth muscle (TSM) cells. Regional (approximately 1.5 microm3) and global (whole cell) changes in [Ca2+]i were measured in fluo-3 loaded TSM cells using real-time confocal microscopy. Regional responses appeared as propagating [Ca2+]i oscillations whereas global responses reflected the spatiotemporal integration of these regional responses. Within a region, [Ca2+]i oscillations were 'biphasic' with initial higher frequencies, followed by slower steady-state oscillations. With increasing ACh concentration, the peak (maximum value relative to 0 nM) of regional [Ca2+]i oscillations remained relatively constant, whereas both frequency and propagation velocity increased. In contrast, the global spatiotemporal integration of the regional oscillatory responses appeared as a concentration-dependent increase in peak as well as mean cellular [Ca2+]i. We conclude that the significance of ACh-induced [Ca2+]i oscillations lies in the establishment of mean [Ca2+]i level for slower Ca2+-dependent physiological processes via modulation of oscillation frequency and propagation velocity. PMID- 11007129 TI - Norepinephrine acts on the KATP channel and produces different effects on [Ca2+]i in oscillating and non-oscillating HIT-T15 cells. AB - Norepinephrine (NE) is an inhibitor of insulin secretion that acts, in part, by decreasing intracellular free calcium ([Ca2+]i). We examined the effects of NE on [Ca2+]i in individual HIT-T15 cells loaded with indo 1. Cells were categorized as oscillators or non-oscillators on the basis of the pattern of the calcium response to glucose and the effect of NE on [Ca2+]i was subsequently measured in each cell. NE caused a simple decrease in [Ca2+]i in nonoscillators. In oscillators, NE decreased the amplitude and frequency of the oscillations. Furthermore, the duration of the NE effect in oscillators was longer than in non oscillators. NE did not affect the rise in [Ca2+]i elicited by depolarizing concentrations of 20 mM or 35 mM KCl alone, or in the presence of 20 mM KCl, 100 microM diazoxide, and 10 mM glucose. In other experiments, NE had no effect on [Ca2+]i when the KATP channels were fully clamped with diazoxide or tolbutamide. We conclude that the action of NE to decrease [Ca2+]i in both oscillators and non oscillators is mediated via activation of the KATP channel. Despite this common mechanism, NE exerts different effects on oscillating and non-oscillating cells. PMID- 11007130 TI - Effects of elevated cytoplasmic calcium and protein kinase C on endoplasmic reticulum structure and function in HEK293 cells. AB - In human embryonic kidney (HEK) cells stably transfected with green fluorescent protein targeted to the endoplasmic reticulum (ER), elevation of intracellular Ca2+ ([Ca2+]i) altered ER morphology, making it appear punctate. Electron microscopy revealed that these punctate structures represented circular and branched rearrangements of the endoplasmic reticulum, but did not involve obvious swelling or pathological fragmentation. Activation of protein kinase C with phorbol 12-myristate 13-acetate (PMA), prevented the effects of ionomycin on ER structure without affecting the elevation of [Ca2+]i. These results suggest that protein kinase C activation alters cytoplasmic or ER components underlying the effects of high [Ca2+]i on ER structure. Treatment of HEK cells with PMA also reduced the size of the thapsigargin-sensitive Ca2+ pool and inhibited Ca2+ entry in response to thapsigargin. Thus, protein kinase C activation has multiple actions on the calcium storage and signalling function of the endoplasmic reticulum in HEK cells: (1) reduced intracellular Ca2+ storage capacity, (2) inhibition of capacitative Ca2+ entry, and (3) protection of the endoplasmic reticulum against the effects of high [Ca2+]i. PMID- 11007131 TI - Evaluation of the effectiveness of a specialist nurse in the management of inflammatory bowel disease (IBD). AB - OBJECTIVE: To determine the effect of a specialist nurse on the management outcome of patients with inflammatory bowel disease (IBD). DESIGN: Audit of the management of a cohort of patients in the year prior to the employment of the specialist nurse and the year immediately after. SUBJECTS: 339 patients, both male and female, with either Crohn's disease or ulcerative colitis, resident in the Cambridge health district. SETTING: Addenbrooke's Hospital NHS Trust Outpatient Centre. MAIN OUTCOME MEASURE: Health status was measured by blood tests (C-reactive protein, albumin and haemoglobin) throughout the year, symptom indices, number of clinic attendances, admissions to hospital and length of stay. Quality of life was measured via a postal questionnaire. RESULTS: Hospital visits were reduced from 1377 to 853 (38% reduction) and in-patient length of stay measured in bed-days from 516 to 417 (19% reduction). The number of patients in remission increased from 63 to 69%. Patient satisfaction improved in key areas, in particular, access to information on IBD and advice on avoidance of illness and maintaining health. Of a total of 251 calls to the telephone helpline, only 19 patients were referred for a medical opinion and five patients required hospital admission. CONCLUSION: The IBD nurse specialist is a valuable and cost effective member of the gastroenterology team. PMID- 11007132 TI - Passive smoking in patients with inflammatory bowel disease: an Israeli multicentre case-control study. AB - BACKGROUND: The association between smoking and inflammatory bowel disease (IBD) is well established. There are, however, no large scale studies of passive smoking in inflammatory bowel disease and this has never been surveyed in the Jewish population of Israel. AIM: To study the passive smoking exposure of Jewish IBD patients in Israel in a large scale multicentre study. METHODS: Patients with established IBD, aged 18-70 years, were interviewed regarding smoking and other habits. Two controls, one clinic and one neighbourhood, matched by age, sex, community group, and education, were sought for each subject. RESULTS: Five hundred and thirty-four patients (273 ulcerative colitis (UC) and 261 Crohn's disease (CD)), 478 clinic controls and 430 community controls were interviewed. There were no significant differences in the passive smoking habits between IBD patients and their controls. Fifty-one percent of UC patients, 50% of the clinic controls and 58% of the community controls were exposed to passive smoking at home (NS); similar results were found among CD patients (50%, 55% and 56%, respectively). When a quantitative exposure index was used UC patients were significantly less exposed to passive smoking than were their community controls (7.46 +/- 8.40 vs 9.36 +/- 9.46, n = 229, P< 0.031). There was no difference in the exposure to passive smoking among CD patients and their controls. No differences in exposure to passive smoking were found when UC patients who had never smoked were compared with their controls. When the quantitative index was used 'never-smoked' CD patients tended to be less exposed to passive smoking at home than their community controls (5.40 +/- 7.60 vs 8.04 +/- 8.72, P < 0.05). CONCLUSION: There is a lack of association between passive smoking and IBD in Jewish patients in Israel. When a quantitative exposure index was used UC patients were found to be less exposed to passive smoking than their community controls. PMID- 11007133 TI - Symptoms discriminate irritable bowel syndrome from organic gastrointestinal diseases and food allergy. AB - BACKGROUND: The value of specific gastrointestinal symptoms in discriminating irritable bowel syndrome (IBS) from organic disease has been documented. In contrast, there have been few attempts to identify symptoms that discriminate irritable bowel syndrome from food allergy, despite similarities in their respective symptom complexes. We aimed to investigate the value of symptoms in discriminating irritable bowel syndrome from organic disease and food allergy. METHODS: Subjects (n = 288) were recruited from consecutive patients presenting to the Internal Medicine, Gastroenterology and Allergy Units in Chieti. Patients completed the validated Bowel Disease Questionnaire (BDQ) prior to an independent diagnostic evaluation, which included endoscopy when appropriate. Food allergy was diagnosed using a 2-week elimination diet, followed by a placebo-controlled food challenge test, a skin prick test and serum RAST for specific IgE for suspected foods or additives. The results of the BDQ were not considered in formulating a diagnosis. In total, 99 patients were diagnosed with the IBS, 79 patients were diagnosed with organic disease and 22 patients were diagnosed with food allergy. A further 88 patients with extraintestinal allergies were included as a control group. RESULTS: Based on logistic regression analysis, six symptom items discriminated IBS from organic disease, while five symptoms discriminated patients with IBS from control subjects. A diagnosis of IBS compared to organic disease was positively associated with straining on defaecation (P=0.0001), diarrhoea (P=0.001) and abdominal bloating (P=0.01), but was negatively associated with pain in the upper abdomen (P=0.0004), reflux (P=0.0001) and appetite loss (P=0.004). A diagnosis of IBS compared to extraintestinal allergy was positively associated with pain relieved by bowel movement (P=0.0001), pain in the lower abdomen (P=0.0006), pain in both the upper and lower abdomen (P=0.003), frequent pain (P=0.001) and abdominal bloating (P=0.0009). In comparison between IBS and food allergy patients, a diagnosis of IBS was positively associated with pain in the lower abdomen (P=0.001), pain relieved by bowel movements (P=0.001), frequent pain (P=0.02) and abdominal bloating (P=0.03). CONCLUSION: Symptoms appear to be useful for discriminating IBS from organic gastrointestinal disease and food allergy. PMID- 11007134 TI - Chondrex (YKL-40), a potential new serum fibrosis marker in patients with alcoholic liver disease. AB - OBJECTIVES: Chondrex (YKL-40) is a mammalian member of a protein family that includes bacterial chitinases. The pattern of its expression in certain tissues such as human liver or cartilage suggests a function in remodelling or degradation of extracellular matrix. The purpose of this study was to assess whether circulating YKL-40 might be a serum fibrosis marker in alcoholics. METHODS: Plasma YKL-40 was determined in 146 consecutive heavy drinkers (106 men, 40 women; mean age, 49.2 +/- 9.0 years). Liver biochemical parameters and serum fibrosis markers such as hyaluronate were also measured. Fibrosis and inflammation in liver biopsy were evaluated using a semi-quantitative scoring system. RESULTS: Plasma YKL-40 increased in parallel with the severity of fibrosis (P<0.00001). YKL-40 also increased in the presence of hepatic inflammation (P<0.01). Receiver operating characteristic curves of Chondrex revealed that a threshold of 330 microg/l gave a specificity of 88.5%; however, the sensitivity was only 50.8%. Only 11.5% of patients without severe fibrosis displayed a Chondrex plasma level above this threshold. A positive correlation was found between Chondrex and hyaluronate (r=0.40, P<0.0001), and a negative correlation was shown between Chondrex and the prothrombin index (r=-0.37, P<0.0001). CONCLUSIONS: The severity of liver fibrosis is associated with elevated circulating Chondrex levels. The overlap in YKL-40 values prevents use of Chondrex in a screening programme. High levels of Chondrex (above 330 microg/l) are predictive of severe liver fibrosis. Increased plasma YKL-40 may reflect the remodelling of liver fibrosis in alcoholics. PMID- 11007135 TI - The impact of chronic hepatitis B viral infection on gastrointestinal motility. AB - OBJECTIVES: Disturbed gastrointestinal (GI) motility probably exists in alcoholic cirrhotic patients; however, the influence of chronic hepatitis B virus (HBV) infection on GI motility remains unknown. The purpose of this prospective study was to determine the impact of chronic HBV infection on human GI transit, and to explore the possible patient factors modulating GI motility. METHODS: We used a non-invasive hydrogen breath test measuring the oro-caecal transit time (OCTT) to assess the GI motility in 45 asymptomatic HBV carriers, 26 patients with chronic hepatitis B, 23 patients with HBV-related liver cirrhosis, and 45 age- and sex matched healthy volunteers. Their clinical symptoms and various blood parameters, such as platelet count, prothrombin time, etc. were recorded. Plasma substance P, nitrate/nitrite and endothelin-1 levels were also measured. RESULTS: The OCTTs in controls, HBV carriers, chronic hepatitis B and liver cirrhosis patients were (mean +/- SEM) 78.4 +/- 5.8, 80.9 +/- 4.2, 93.9 +/- 8.8 and 106.5 +/- 12.4 min, respectively. The OCTT was delayed in patients with HBV-related liver cirrhosis compared to that of controls (P=0.039). Among the cirrhotic patients, presentation with ascites delayed OCTT (145.7 +/- 27.2 versus 91.3 +/- 11.9 min, P=0.039). Neither Child- Pugh grade, portal hypertension, various blood parameters, plasma substance P, nitrate/nitrite or endothelin-1 levels had any influence on OCTT. CONCLUSIONS: HBV infection alone does not alter GI motility, whereas the patients with liver cirrhosis may have delayed GI motility. Ascites is most likely a factor responsible for the delayed GI transit among chronic HBV infected subjects. PMID- 11007136 TI - Decrease of regional cerebral blood flow in liver cirrhosis. AB - OBJECTIVE: Alterations of regional cerebral blood flow (rCBF) in subjects with liver cirrhosis have not been fully evaluated. We evaluated quantitative changes in rCBF using single photon emission computed tomography (SPECT). METHODS: Twenty eight Japanese patients with liver cirrhosis were enrolled in this study. None of them exhibited advanced hepatic encephalopathy at the time of examination. The cause of liver cirrhosis was viral infection in 26 patients; the cause was unknown in two patients. Child-Pugh classification of the patients was as follows: Group A, 12 patients; Group B, 12 patients; and Group C, four patients. The control group consisted of 25 age-matched healthy subjects. Radionuclide angiography was performed by rapid injection of Tc-99m ethyl cysteinate dimer (ECD) (740 MBq) via the right cubital vein, and then SPECT brain images were taken. Using the Patlak graphical method, rCBF values (ml/100 g per min) were calculated in the frontal, parietal, temporal and occipital lobes and cerebellum on SPECT images. RESULTS: The rCBF values were lower in cirrhotic patients than in controls, i.e. by 15% in the frontal lobe, by 12% in the parietal lobe, by 10% in the temporal and occipital lobes, and by 7% in the cerebellum. They decreased concomitantly with the severity of liver disease. A significant negative correlation was noted between rCBF values and Child-Pugh score in the frontal (P<0.01), parietal (P<0.05) and occipital lobes (P<0.01). rCBF values of each region were not correlated with age or with results of neuropsychological test. The degree of association between rCBF values and results of laboratory examination was generally poor. CONCLUSION: Patients with liver cirrhosis without advanced encephalopathy showed widespread reduction in rCBF; this reduction was particularly evident in the frontal lobe. Tc-99m ECD SPECT may be useful for evaluating cerebral functional changes in patients with liver cirrhosis. PMID- 11007137 TI - Liver injury due to tetrabamate (Atrium): an analysis of 11 cases. AB - BACKGROUND: Tetrabamate (Atriums), widely used in the treatment of tremor and ethanol-withdrawal symptoms, has been incriminated as a potential cause of reversible acute hepatitis. OBJECTIVE: We report here on 11 patients who experienced tetrabamate-related liver injury, in order to evaluate their clinical, histopathological and evolutive features. PATIENTS AND METHODS: Between 1987 and 1998, 34 cases of tetrabamate-associated acute hepatitis were spontaneously reported to the regional pharmacovigilance center of Lyon. Eleven cases were considered probably to be drug-induced. RESULTS: There were three males and eight females aged 31-82 years (mean, 57 years). The duration of treatment ranged from 33 to 206 days, and indication for treatment was depressive disorders, tremor or prevention of alcohol withdrawal symptoms. Clinical symptoms were asthenia (n = 9), jaundice (n = 3) and/or diffuse rash (n = 3). The pattern of liver injury was cytolytic (n = 10) or cholestatic (n = 1). Three patients presented biological features of hepatic failure. A percutaneous liver biopsy was performed in six patients. Histological examination of the liver specimen showed a large spectrum of lesions: massive hepatocellular necrosis (n = 1), centrilobular and nonconfluent hepatocyte necrosis (n = 2), intracellular cholestasis (n = 3), and granulomatous hepatitis (n = 1). Tetrabamate was discontinued in all patients. In seven patients, a complete recovery was observed 3 weeks to 3 months after drug withdrawal. Two patients, despite a rapid improvement of liver function tests, died from unrelated causes. The remaining two patients died from irreversible hepatic failure. CONCLUSIONS: Our data strongly suggest that tetrabamate may induce acute liver injury, which may eventually result in life-threatening liver failure. PMID- 11007138 TI - Soda bread provocation test for subjects with transient serology for coeliac disease 3 years after a population screening survey. AB - BACKGROUND: We have previously reported that IgA antigliadin antibodies (IgA-AGA) in the majority of healthy subjects are transient and do not indicate enteropathy, and also that an increased intake of gluten in the form of soda bread may be part of the explanation for this phenomenon. OBJECTIVE: The aim of the study was to determine whether gluten challenge with soda bread in subjects with transiently positive IgA-AGA could induce a significant titre of IgA-AGA. DESIGN: Food challenge study. METHODS: All subjects with positive IgA-AGA on screening at the time of the MONICA project in 1991 (T0) who developed negative serology at 3-year follow-up (T1) were invited to participate in a 'soda bread challenge' (1 loaf per day) for 1 month. Analysis of food intake was carried out prior to the challenge (T2) and compared to the analysis at the time of screening (T0). IgA-AGA and IgA antiendomysial antibodies (EMA) were checked pre- (T2) and post-challenge (T3). RESULTS: Ten subjects agreed to participate. Quantities of food ingested for the various categories did not differ significantly from T0 to T2. IgA-AGA titres did not differ significantly from T1 to T2 (20.2 versus 35.0, P=0.085). Mean IgA-AGA titres rose significantly between T2 and T3 (35.0 versus 40.3, P=0.005), although none of the subjects developed a significant titre of IgA-AGA. None of the subjects were positive for IgA-EMA. CONCLUSIONS: Intake of soda bread does not appear to be an important explanation as to why subjects may have a transient rise in IgA-AGA titre since none of the subjects developed a significant titre of IgA-AGA. PMID- 11007139 TI - Evidence for existence of coeliac disease autoantigens apart from tissue transglutaminase. AB - BACKGROUND: The pathogenesis of coeliac disease (CD) and of dermatitis herpetiformis (DH) is strongly associated with production of autoantibodies, defined by indirect immunohistology. Recently, tissue transglutaminase (tTG) was identified as a prominent autoantigen. It would be important to investigate if further molecules apart from tTG are involved in autoimmunity. METHODS: Tissue sections of human foetal intestine were used to compare the distribution of tTG with the autoantibody binding patterns of 14 sera samples from patients with CD or DH. Double label experiments were performed using monoclonal as well as polyclonal tTG antibodies (anti-tTG) and patient sera. The staining was investigated by using conventional light and confocal laser scanning microscopy. RESULTS: Most autoantibody binding sites were matched by tTG. Further, the binding of autoantibodies could be inhibited by preincubation with monoclonal anti-tTG. However, in nine serum samples (64%) autoantibody staining suggested a few distinct binding sites apart from tTG. In three sera (21 %) autoantibody binding fibres were detected which definitely did not match monoclonal anti-tTG signals. Distinctly stained fibres were confirmed by applying polyclonal anti tTG. This indicates the existence of autoantigenic epitopes not related to tTG. PMID- 11007140 TI - Idiopathic fibrosing pancreatitis and Crohn's disease: an interesting association. AB - Idiopathic fibrosing pancreatitis is an uncommon condition in children and adolescents of unknown aetiology. This syndrome has been reported in 36 cases so far. To our knowledge none of these cases was definitively associated with Crohn's disease. In this report we describe a young female patient who developed Crohn's disease of the colon 5 years after having been diagnosed with idiopathic fibrosing pancreatitis. The differential diagnosis between this syndrome associated with Crohn's disease and pancreatic Crohn's disease or fibrosing colonopathy, an entity related to pancreatic enzyme therapy, is discussed. PMID- 11007141 TI - A case of haemosuccus pancreaticus. AB - Haemosuccus pancreaticus (Wirsungorrhagia or pseudohaemobilia) is a rare complication of chronic pancreatitis. We describe a 48-year-old patient with alcohol-induced chronic calcific pancreatitis and recurrent episodes of severe upper gastrointestinal bleeding but without abdominal pain. Upper gastrointestinal endoscopy revealed fresh blood oozing from the ampulla of Vater. No pseudoaneurysms or pseudocysts were detected by arteriography or computerized tomography. The bleeding was attributed to pancreatic lithiasis. Following conservative treatment, there was no evidence of recurrence during a 24-month follow-up period. In conclusion, although a rare occurrence, haemosuccus pancreaticus should be considered in the differential diagnosis of all cases of obscure upper gastrointestinal bleeding in patients with chronic pancreatitis, whether or not accompanied by pain. A highly suggestive clinical history or X-ray findings and an endoscopic visualization of blood coming from the ampulla of Vater may suffice for the diagnosis, thus avoiding diagnostic and therapeutic errors. When haemosuccus pancreaticus occurs in patients without pseudoaneurysms or pseudocysts, it can be treated conservatively, thus obviating the need for pancreatectomy or arteriographic embolization. PMID- 11007142 TI - Development of small hepatocellular carcinoma 80 months after clearance of hepatitis C virus with interferon therapy. AB - We describe a patient who had a complete and sustained response to interferon (IFN) therapy for chronic hepatitis C but developed small hepatocellular carcinoma (HCC) 80 months later. A 55-year-old Japanese man with hepatitis C virus (HCV) infection and histological features of chronic active hepatitis was treated with recombinant IFN alpha-2a, 9,000,000 U daily for 2 weeks followed by three times a week for 22 weeks. He successfully responded to IFN therapy with a normalization of serum alanine aminotransferase and continuous disappearance of serum HCV-RNA. However, 80 months after the cessation of IFN therapy, the patient's alpha-fetoprotein (AFP) level became elevated for the first time and HCC, 12 mm in diameter, was detected by routine ultrasonographic screening. Laparotomy revealed a small HCC with no metastasis, and the nontumorous liver demonstrated chronic inactive hepatitis. This case indicates the need for careful follow-up using ultrasonography and AFP testing for at least 7 years after completing IFN therapy in all patients with chronic hepatitis C, even if the patients have a complete response to the therapy. PMID- 11007143 TI - Gastric adenocarcinoma in a patient with X-linked agammaglobulinaemia. AB - Patients with primary immunodeficiencies are at high risk for developing haematological malignancies and, to a lesser degree, carcinoma. We report a patient with ascertained X-linked agammaglobulinaemia who developed a gastric carcinoma involving the distal part of the stomach associated with chronic atrophic gastritis and intestinal metaplasia. These latter conditions are considered to be precursor conditions and the role of chronic infections is likely. Patients with X-linked agammaglobulinaemia, as with other primary immunodeficiencies, could benefit from regular gastrointestinal evaluation, leading to early diagnosis and treatment of carcinoma. PMID- 11007144 TI - Multivisceral eosinophilic fibrosis: a new clinical presentation. AB - We describe a patient who was referred to us with the diagnosis of pancreatic cancer but who had eosinophilic fibrosis of the pancreas and other organs, including the sub-mandibular salivary glands, retro-orbital tissue, liver, kidneys, and surrounding the abdominal aorta. He had no pain or other symptoms. After treatment with methylprednisolone, all lesions disappeared and now he seems to be cured of this apparently immune-mediated disorder. To our knowledge, involvement of all these particular organs by eosinophilic fibrosis in the absence of symptoms has never before been described. PMID- 11007145 TI - Cushing's syndrome in an 81-year-old patient treated with budesonide and amiodarone. AB - This is the first report of Cushing's syndrome under oral budesonide treatment. An 81-year-old man known for paroxysmal atrial fibrillation and chronic renal insufficiency, treated with 6 mg budesonide for collagenous colitis, developed Cushing's syndrome under co-administration of amiodarone. The Cushing's syndrome disappeared after discontinuation of the amiodarone treatment. Metabolism of the two medications by hepatic cytochrome P 450 3A may explain the development of Cushing's syndrome. PMID- 11007146 TI - Epidemiology and interventiaonl studies. Helicobacter pylori and gastric cancer: progression/regression of histopathological lesions. PMID- 11007147 TI - Transdermal slow-release long-acting isosorbide dinitrate for 'nutcracker' oesophagus: an open study. PMID- 11007148 TI - English language use as a risk factor for smoking initiation among Hispanic and Asian American adolescents: evidence for mediation by tobacco-related beliefs and social norms. AB - Acculturation increases the risk of smoking among Hispanic and Asian American adolescents, but the underlying mechanisms are not understood. This study examined associations between English language use and smoking among 4,167 Hispanic and 2,836 Asian American adolescents in California. Potential mediators were assessed, including access to cigarettes, perceived consequences, friends' smoking, cigarette offers, refusal self-efficacy, and prevalence estimates of peer smoking. English language use was associated with increased risk of lifetime smoking in both groups. This association became nonsignificant after access, perceived consequences, friends' smoking, and offers were controlled for. The acculturation process (as indicated by English language use) may be associated with smoking-related psychosocial variables, which may lead to an increased risk of experimentation with smoking. PMID- 11007149 TI - Sociodemographic and psychological predictors of changes in dietary fat consumption in adults with high blood cholesterol following counseling in primary care. AB - The associations between sociodemographic variables, psychological factors, and changes in dietary fat consumption over 4 months were assessed in a randomized controlled trial of behavioral counseling versus standard advice. Patients were 141 men and 150 women, with an average age of 52.1 years and total cholesterol level of 7.27 mmol/l (278 mg/dl). Smokers, younger patients, and those with greater body mass index had higher fat intake at baseline. Behavioral counseling led to greater reductions in fat intake than did standard advice. Self-efficacy and ratings of benefits of low-fat diets were related to fat consumption at baseline, and changes in these measures were correlated with changes in fat intake. Family support, baseline anticipated regret, and (for the behavioral counseling group only) baseline behavioral intentions predicted reductions in fat intake. The results indicated that psychosocial variables associated cross sectionally with fat consumption do not necessarily predict change and that factors involved in the process of change and the prediction of change need to be differentiated. PMID- 11007150 TI - Gender-linked personality traits predict mental health and functional status following a first coronary event. AB - Three gender-linked traits were examined with respect to adjustment to a coronary event: agency, a focus on the self; communion, a focus on others; and unmitigated communion, an extreme focus on others to the exclusion of the self. Participants (n = 65) were interviewed 1 week and 4 months after a 1st coronary event. Hypotheses were that agency should predict improved health, communion should be unrelated to health, and unmitigated communion should predict worse health over time. Outcomes included depression, anxiety, and well-being (as measured by the Profile of Mood States; D. McNair, M. Lorr, & L. Droppleman, 1971); mental and physical functioning (SF-36; J. E. Ware, K. K. Snow, M. Kosinski, & B. Gandek, 1993); and cardiac symptoms. Results confirmed hypotheses. In addition, unmitigated communion was linked with poor health behavior and negative social interactions, which partly explained the link of unmitigated communion with depression and cardiac symptoms. PMID- 11007151 TI - Job strain and risk indicators for cardiovascular disease in young female nurses. AB - This study examined the possible effects of job demands, decision latitude, and job-related social support on risk indicators for cardiovascular disease (CVD) in 165 female nurses. Job strain was measured with the Job Content Questionnaire; CVD risk was measured with insulin, total cholesterol, triglyceride (TG), high density lipoprotein cholesterol (HDL-C), fibrinogen, tissue-type plasminogen activator (tPA) antigen, tPA activity, plasminogen activator inhibitor-1 antigen, and blood pressure. Multivariate analysis of covariance and regression analyses revealed no effects of either job strain or social support on these risk indicators. All risk indicators deteriorated with age and body mass index. Oral contraceptive use improved fibrinolytic potential and increased HDL-C but had adverse effects on TG levels. Results suggest that in healthy young women job strain is not associated with an unfavorable metabolic or fibrinolytic risk profile. PMID- 11007152 TI - Psychological response styles and cardiovascular health: confound or independent risk factor? AB - Using results from 2 large cardiovascular studies, the authors examined the utility of treating psychological response styles as confounds (e.g., factors undermining relationships with other self-report variables) versus distinct personality traits in the prediction of cardiovascular health. Study 1 consisted of a 3-year prospective study of ambulatory blood pressure levels in healthy adults (N = 125). Study 2 comprised a 12-week drug treatment program for ischemic heart disease patients (N = 95). Participants completed measures of psychological factors and self-deception and impression management in each study. Results consistently favored using response styles as direct predictors. Self-deception scores predicted elevated 3-year diastolic and systolic blood pressure changes in Study 1 and poorer treatment outcomes in Study 2. Statistically controlling for response style effects within the psychological factors generally did not improve predictions. These findings argue against the conceptualization of response styles as stylistic confounds. PMID- 11007153 TI - Motivation and dietary self-care in adults with diabetes: are self-efficacy and autonomous self-regulation complementary or competing constructs? AB - This study examined constructs drawn from social-cognitive theory (A. Bandura, 1986) and self-determination theory (E. L. Deci & R. M. Ryan, 1985, 1991) in relation to dietary self-care and life satisfaction among 638 individuals with diabetes. A motivational model of diabetes dietary self-care was proposed, which postulates direct links between self-efficacy/autonomous self-regulation, and adherence/ life satisfaction. Structural equation modeling showed that both self efficacy and autonomous self-regulation were associated with adherence (betas = .54 and .21, respectively) and with life satisfaction (betas = .15 and .34, respectively). Constraint analyses confirmed that self-efficacy was significantly more associated with adherence, whereas autonomous self-regulation was significantly more associated with life satisfaction. According to the model, interventions for dietary self-care and life satisfaction should focus on increasing self-efficacy and autonomous self-regulation. PMID- 11007154 TI - Structure of outcome beliefs in condom use. The Project RESPECT Study Group. AB - To study the structure of beliefs about condom use outcomes, the authors derived and tested 4 psychosocial hypothetical models: (a) a 2-factor model of the personal and social outcomes of condom use; (b) a 2-factor model of the pros and cons of the behavior; (c) a 3-factor model (i.e., physical, self-evaluative, and social) of outcome expectancies; and (d) a thematic 4-factor model of the protection, self-concept, pleasure, and interaction implications of the behavior. All 4 models were studied with a confirmatory factor analysis approach in a multisite study of 4,638 participants, and the thematic solution was consistently the most plausible. Self-concept and pleasure were most strongly associated with attitudes toward using condoms, intentions to use condoms, and actual condom use, whereas protection and interaction generally had little influence. PMID- 11007155 TI - A psychosocial model of sun protection and sunbathing in young women: the impact of health beliefs, attitudes, norms, and self-efficacy for sun protection. AB - A psychosocial model of sun protection and sunbathing as distinct behaviors was developed on 202 young Caucasian women and replicated in an independent sample (n = 207). Proximal outcomes were intention to sun protect and intention to sunbathe; distal outcomes included sun protection and sunbathing behavior measured 5 months later. Objective risk for skin cancer plus 4 classes of psychosocial variables (sun-protective health beliefs, self-efficacy for sun protection, attitudes toward sunbathing, and norms for sunbathing and sun protection) served as predictors. Sun-protective norms and self-efficacy for sun protection predicted only intention to sun protect; sunbathing norms predicted only intention to sunbathe. Susceptibility and advantages of tanning predicted both intention constructs, which, in turn, predicted behavior. These findings distinguish sun protection from sunbathing and provide a basis for intervention design. PMID- 11007156 TI - Social-cognitive determinants of nutrition behavior among supermarket food shoppers: a structural equation analysis. AB - A social-cognitive model of nutrition behavior (A. Bandura, 1997) was tested using structural equation modeling of data from 307 food shoppers recruited from 5 supermarkets in Southwest Virginia. The shoppers were participating in the baseline phase of an ongoing nutrition promotion program. As part of the evaluation, data were collected on the self-efficacy and outcome-expectations components of social-cognitive theory as well as on food purchases and intake. The structural analyses presented here support the social-cognitive model. Self efficacy, physical outcome expectations, age, socioeconomic status, and number of children were important predictors of nutrition behavior among shoppers. Implications for interventions using social-cognitive variables to improve the nutritional content of food purchases and intake are discussed. PMID- 11007157 TI - Social-cognitive predictors of health behavior: action self-efficacy and coping self-efficacy. AB - The effects of social-cognitive variables on preventive nutrition and behavioral intentions were studied in 580 adults at 2 points in time. The authors hypothesized that optimistic self-beliefs operate in 2 phases and made a distinction between action self-efficacy (preintention) and coping self-efficacy (postintention). Risk perceptions, outcome expectancies, and action self-efficacy were specified as predictors of the intention at Wave 1. Behavioral intention and coping self-efficacy served as mediators linking the 3 predictors with low-fat and high-fiber dietary intake 6 months later at Wave 2. Covariance structure analysis yielded a good model fit for the total sample and 6 subsamples created by a median split of 3 moderators: gender, age, and body weight. Parameter estimates differed between samples; the importance of perceived self-efficacy increased with age and weight. PMID- 11007158 TI - Conscientiousness, perceived risk, and risk-reduction behaviors: a preliminary study. AB - Participants (N = 343) from an Oregon community completed surveys at baseline, 3 months, and 12 months to assess personality, the perceived health risk of radon in combination with smoking, and changes in smoking behavior. Conscientiousness predicted instituting a more restrictive household smoking rule (p < .01), and perceived risk predicted reduction in cigarettes smoked per day for men (p < .001). Perceived risk predicted a reduction in the proportion of cigarettes smoked in the home for those who had high (p < .05) but not low or moderate levels of Conscientiousness, a dimension in one personality model. The results demonstrate the importance of Conscientiousness in the prediction of health behavior, particularly behavior that affects others as well as oneself. PMID- 11007159 TI - Hypomethylation of the retrotransposon LINE-1 in malignancy. PMID- 11007160 TI - Levels of beta-human chorionic gonadotropin in cerebrospinal fluid of patients with malignant germ cell tumor can be used to detect early recurrence and monitor the response to treatment. AB - BACKGROUND: Tumor marker-producing germ cell tumors of the central nervous system are malignant and require radiation and/or chemotherapy. Although serum beta human chorionic gonadotropin (hCG) has been used to monitor the course of treatment, the levels of beta-hCG in the cerebrospinal fluid (CSF) have not been measured routinely in the clinic. To determine whether they can be used to evaluate parameters of tumor status, such as progression or response to therapy, levels of beta-hCG in the serum and CSF of patients with germ cell tumors were studied. METHODS: Fifty-four paired samples of CSF and serum were taken from seven patients with germ cell tumor and their beta-hCG levels were measured. Beta hCG was negative in both serum and CSF in 11 instances and the levels in the other 43 paired samples were analyzed for any correlation or relationship to therapy. They were also compared with the clinical courses. RESULTS: The mean CSF beta-hCG level was 11.5 mIU/ml, which was significantly higher than the level in serum (3.5, p = 0.002). In all the paired samples except for one time point, the level in CSF was higher than that in serum. Out of 43 instances where the beta hCG level in CSF was elevated, the level in serum was elevated in only 16 (37.2%). Among cases of recurrent malignant germ cell tumor, there were nine instances of recurrence or progression despite therapy. In all five instances where beta-hCG CSF levels were measured, the levels were elevated prior to any increase or detectability of the serum values. CONCLUSION: It seems likely that the level of beta-hCG in CSF is a good marker for monitoring tumor recurrence or evaluation of treatment results. PMID- 11007161 TI - Heterogeneous response patterns of alveolar macrophages from patients with lung cancer by stimulation with interferon-gamma. AB - BACKGROUND: Macrophages are considered to play an important role in the host defense against malignant tumors. In this study, cytotoxic activity of alveolar macrophages (AM) derived from 32 patients with lung cancer was investigated. METHODS: AM were aseptically obtained by lavage from resected lung and subsequently tested for cytolytic activity against QG56, a lung squamous cell line, following treatment with recombinant interferon-gamma (IFN-gamma). RESULTS: In seven patients (21.9%), AM showed no cytotoxicity even though AM were incubated with IFN-gamma. In 20 (62.5%), AM showed substantial cytotoxicity in response to IFN-gamma in a dose-dependent manner. In the other five (15.6%), relatively strong cytotoxicity was observed even without preincubation with IFN gamma. Such a heterogeneous profile of the cytotoxicity of AM might be a reflection of various activated states of AM since the potential of cytotoxicity and that of IL-1 secretion were almost parallel. Both IFN-gamma dependent and independent cytotoxicity were partially blocked either by anti-tumor necrosis factor-alpha (TNF-alpha) antibody or by the inhibitor of nitric oxide synthesis. However, those activities were completely abrogated by both treatments. Since the supernatant of AM culture exhibited TNF-alpha-mediated but not NO-mediated cytolysis, TNF-alpha could mediate a bystander killing whereas NO acts in close contact with tumor cells. CONCLUSION: The AM have anti-tumor cytotoxicity in lung cancer although the cytolytic potential is heterogeneous and that the tumor lysis by AM is mediated by both TNF-alpha and NO production. PMID- 11007162 TI - Characterization of intracellular cytokine profile of CD4(+) T cells in peripheral blood and tumor-draining lymph nodes of patients with gastrointestinal cancer. AB - BACKGROUND: Analysis of serum cytokine levels has shown that cancer-bearing hosts have lower levels of IL-2 and IFN-gamma, suggesting that Th1-type immunity is impaired by cancer. However, the mechanisms of the Th1 dysfunction are not clearly understood. METHOD: The frequencies of Th1 cells in CD4(+) helper T cells were evaluated with an intracytoplasmic cytokine staining method in peripheral blood lymphocytes (PBL) and lymph node lymphocytes (LNL) of patients with gastrointestinal cancer. RESULTS: Activation of lymphocytes with PMA + Ionomycin induced the expression of IL-2 and IFN-gamma in each lymphocyte population. Compared with PBL of non-malignant donors, PBL in cancer patients contained significantly lower frequencies of CD4(+) T cells that produced IL-2 and IFN gamma. LNL in cancer patients also contained lower levels of IL-2- and IFN-gamma producing CD4(+) T cells, although the percentages did not show significant differences from those of PBL in the same patients. CONCLUSION: Our data suggest that suppression of Th1 cytokine in cancer patients is, at least in part, due to the decreased frequency of Th1 cells with CD4(+) phenotype. PMID- 11007163 TI - Hypomethylation of LINE1 retrotransposon in human hepatocellular carcinomas, but not in surrounding liver cirrhosis. AB - BACKGROUND: Cytosine methylation of LINE1 (L1) elements, some of which are capable of retrotransposition in human cells, is known to play important roles in transcriptional repression of these retrotransposons. We have previously identified consistent hypomethylation of L1 elements in mouse liver tumors by a genome-wide search technique for aberrant methylations. In this study, we analyzed the methylation status of the L1 elements in human hepatocellular carcinomas (HCCs). METHODS: Nine pairs of an HCC and its surrounding tissue were obtained from clinical cases. Genomic DNA was digested with HpaII, a methylation sensitive restriction enzyme, and hybridized with a probe derived from the promoter region of the L1 elements. RESULTS: Hypomethylation of the L1 elements was detected in eight of the nine HCCs, but never in the surrounding liver tissues, whether or not liver cirrhosis was present. CONCLUSION: Specific occurrence of the hypomethylation of the L1 elements in the HCCs indicated its diagnostic value for malignancy. The hypomethylation could also lead to increased incidence of retrotransposition and resultant genomic instability in HCCs. PMID- 11007164 TI - Essential thrombocythemia transformed to acute myelogenous leukemia with t(3;17)(p24; q12), del(5)(q13q34) after treatment with carboquone and hydroxyurea. AB - In 1991, a 52-year-old woman was diagnosed as having essential thrombocythemia (ET). She was doing well with continuous medication with carboquone (CQ) and subsequently hydroxyurea (HU). However, substantial leukocytosis with leukemic blast cells, anemia and thrombocytopenia developed in 1996. Analysis of peripheral blood showed 4.4 x 10(3)/microl white blood cells with 82% of leukemic blast cells. These blasts showed negative staining with myeloperoxidase by immunostaining, but the myeloperoxidase was positive by electron microscopic analysis. Cytogenetic analysis of bone marrow cells revealed a t(3;17)(p24; q12), del(5)(q13q34). On the basis of these findings, the leukemic blast cells were classified as acute myelogenous leukemia (AML:M0) in the FAB classification. The causative agent, CQ and HU in secondary leukemia from ET and chromosomal abnormality related to ET blastic crisis (BC) are discussed. PMID- 11007165 TI - Primary extracranial meningioma of the foot: a case report. AB - We present a rare case of primary extracranial meningioma in a 36-year-old man, who had a solitary multinodular mass located in the plantar muscle of the foot. The histology of specimens from simple excision was typical of meningioma, showing bland spindle cell proliferation with a whorl pattern. Immunohistochemical analysis demonstrated that the tumor cells showed diffuse and strong positivity for epithelial membrane antigen as well as moderate reactivity for cytokeratin and vimentin. Ultrastructurally, the tumor cells were characterized by thin bipolar cytoplasmic processes and joined by multiple small desmosomes. There were frequent pinocytotic vesicles and a distinct external lamina on the cell surface. These findings suggest that this primary ectopic meningioma, arising in the soft tissue, may have been derived from perineurial cells of the peripheral nerve, but was morphologically distinguishable from perineurioma. Primary extracranial meningioma should be included in the differential diagnosis of soft-tissue spindle cell tumors, especially those of peripheral nerve origin. PMID- 11007166 TI - Cancer incidence and incidence rates in Japan in 1995: estimates based on data from nine population-based cancer registries. The Research Group for Population based Cancer Registration in Japan. PMID- 11007167 TI - Report of the Thirteenth International Symposium of the Foundation for Promotion of Cancer Research: cancer screening-past, present and future. PMID- 11007168 TI - Cancer statistics digest. Pancreas cancer mortality rates by prefectures in Japan. PMID- 11007169 TI - Nutrition and energetics of the canary (Serinus canarius). AB - Canaries appear to be primarily seed-eaters, although there are no reports of their feeding ecology in the wild. In captivity, they are offered seed-based diets, preferring to consume seeds such as canary, rapeseed and millet. The mean daily dry-matter intake ranges from 3 to 4 g, which corresponds to a mean gross energy intake of approximately 70 kJ per bird per day. The efficiency of dietary metabolism is high (0.85), which equates to individual metabolizable energy intakes of 45-75 kJ per bird per day. For a canary of average body weight (22 g) the data can be fitted to a regression equation to predict a requirement of 62 kJ ME per day. This corresponds to published information on the energy requirements of other passerine species, but deviates from the predictive equation for poultry. The digestibility values for protein, fat and carbohydrate are similar to those obtained for the budgerigar, although it is likely that the digestibility coefficient is dependent upon the seed type and alimentary tract lipase and amylase activities. Nutrient requirements of canary chicks have not yet been determined, although recent studies have provided data on the nutrient intakes of developing chicks. The newly-hatched canary chick has a rapid growth rate, achieving 90% of its asymptotic body mass by 11 days of age. Gross energy intake is approximately 3 kJ per day following hatching and by day 10 is equivalent to that of an adult canary. It appears that the protein intake should lie between 16.5 and 21.9% of the diet (as is), with peak intake occurring between 8 and 10 days of age. PMID- 11007170 TI - Kinetic characterization of outer-ring deiodinase activity (ORD) in the liver, gill and retina of the killifish Fundulus heteroclitus. AB - Conversion of T4 to T3 is the first step in TH action and deiodinases are the major determinants of TH tissue availability and disposal. We here report the kinetic characterization of the outer-ring deiodinating (ORD) enzymes in the liver, gill and retina of sea water-adapted killifish, by using both rT3 and T4 as substrates. In liver, by using rT3, we detected a high Km (84 nM) and a low Km (1.3 nM) component with kinetic characteristics similar to mammalian deiodinases DI and DII. In contrast, T4-ORD only generated a low Km (0.5 nM) component. As judged by its Vmax (920 fmol 125I/mg per h) this DII enzyme is very abundant, approximately five and 20 times higher than that found in trout liver and hypothyroid rat, respectively. Kinetic analysis in killifish gill showed only one enzymatic component, with a high rT3 Km (430 nM) and a relatively low Vmax (4.3 pmol 125I/mg per h). Our results in killifish retina show the expression of a T4 low Km (0.6 nM) deiodinase with high cofactor requirements akin to the mammalian DII. The Vmax value for this enzyme is 182 fmol 125I/mg per h, five times lower than the one found in killifish liver, but comparable to that in hypothyroid rat pituitary. The biochemical similarities between fish and mammalian deiodinases could reflect their high conservation during vertebrate evolution and thus their importance in the regulation of thyroid hormone action. PMID- 11007171 TI - Expression of porcine adipocyte transcripts during differentiation in vitro and in vivo. AB - Transcript concentrations for the transcription factors, CCAAT enhancer binding protein beta and alpha (C/EBPbeta and C/EBPalpha), plus the adipocyte characteristic proteins, fatty acid synthase (FAS), glucose transporter 4 (Glut 4), hormone-sensitive lipase (HSL), insulin receptor (InsR), lipoprotein lipase (LPL), and leptin were measured during differentiation of porcine stromal vascular (S/V) cells in vitro. These same transcripts, excluding FAS and InsR, were measured in porcine adipose tissue from birth to 7 weeks of age. In S/V cells, C/EBPbeta and InsR were continuously elevated. At day 0, C/EBPalpha was approximately 20% of the day 9 value. The LPL increased gradually from day 0 to 9, whereas most other transcripts had a lag period of several days. In tissue, C/EBPbeta was substantial at birth and increased gradually. The C/EBPalpha was relatively low at birth and increased at day 17. The LPL and leptin increased continuously. The Glut 4 was low at birth and increased at day 28. The HSL was relatively low at birth, increased at day 10, and plateaued at day 28. Transcripts in porcine S/V cells develop somewhat differently from adipocyte differentiation models established in clonal cells, but the porcine cells represent a model that should be more applicable to pigs. PMID- 11007172 TI - Characterization of (Na+, K+)-ATPase in gill microsomes of the freshwater shrimp Macrobrachium olfersii. AB - To better understand the adaptive strategies that led to freshwater invasion by hyper-regulating Crustacea, we prepared a microsomal (Na+, K+)-ATPase by differential centrifugation of a gill homogenate from the freshwater shrimp Macrobrachium olfersii. Sucrose gradient centrifugation revealed a light fraction containing most of the (Na+, K+)-ATPase activity, contaminated with other ATPases, and a heavy fraction containing negligible (Na+, K+)-ATPase activity. Western blotting showed that M. olfersii gill contains a single alpha-subunit isoform of about 110 kDa. The (Na+, K+)-ATPase hydrolyzed ATP with Michaelis Menten kinetics with K5, = 165+/-5 microM and Vmax = 686.1+/-24.7 U mg(-1). Stimulation by potassium (K0.5 = 2.4+/-0.1 mM) and magnesium ions (K0.5 = 0.76+/ 0.03 mM) also obeyed Michaelis-Menten kinetics, while that by sodium ions (K0.5 = 6.0+/-0.2 mM) exhibited site site interactions (n = 1.6). Ouabain (K0.5 = 61.6+/ 2.8 microM) and vanadate (K0.5 = 3.2+/-0.1 microM) inhibited up to 70% of the total ATPase activity, while thapsigargin and ethacrynic acid did not affect activity. The remaining 30% activity was inhibited by oligomycin, sodium azide and bafilomycin A. These data suggest that the (Na+, K+)-ATPase corresponds to about 70% of the total ATPase activity; the remaining 30%, i.e. the ouabain insensitive ATPase activity, apparently correspond to F0F1- and V-ATPases, but not Ca-stimulated and Na- or K-stimulated ATPases. The data confirm the recent invasion of the freshwater biotope by M. olfersii and suggest that (Na+, K+) ATPase activity may be regulated by the Na+ concentration of the external medium. PMID- 11007173 TI - Trypsin-like enzymes from two morphotypes of the 'living fossil' Triops (Crustacea: Branchiopoda: Notostraca). AB - Trypsin-like enzymes from two morphotypes (here called short and long) of the 'living fossil' Triops of Baja California Sur, Mexico were studied. Adults of both morphotypes were obtained from outdoor static cultures using dry soil from the natural habitats as a source of cysts and culture substrate. Individual and pooled extracts were made from dissected digestive tubes. The effect of pH and temperature on the trypsin activity was studied using N-alpha-benzoyl-DL-Arg-p nitroanilide (BAPNA) as substrate. The highest proteolytic activity was found at the same pH with extracts of both morphotypes. At this pH, there was greater proteolytic activity at a lower temperature with the short morphotype extract than with the long morphotype extract. Substrate-SDS-PAGE zymograms showed bands of activity. Short morphotype extracts produced six bands; five of them were serine proteases of which three were trypsin-like enzymes. Long morphotype extracts revealed eight bands; six of them were serine proteases of which three were trypsin-like enzymes. PMID- 11007174 TI - Characterization of the MTF-1 transcription factor from zebrafish and trout cells. AB - The metal response element (MRE)-binding transcription factor-1, MTF-1, is a zinc responsive protein that controls transcription of metallothionein (MT) genes in many cell types. In addition, MTF-1 is also hypothesized to regulate transcription of a battery of genes involved in the defense against oxidative stress. Manipulating the Zn concentration in the low microM range reversibly modulates the DNA-binding activity of the mammalian MTF-1; this effect is inhibited at low temperature. This report examines the presence and binding properties of MTF-1 in cell lines derived from warm- and cold-water fishes (zebrafish and trout, respectively). We found that both species of fish express MRE-specific binding activities that are immunologically similar to mouse MTF-1. MTF-1-binding from the cells of both species of fish was activated when cells were treated with Zn but not with Cd. Zebrafish cells contained a single isoform of MTF-1 with binding properties similar to mammalian MTF-1. Trout cells, on the other hand, contained two isoforms of MTF-1: MTF-1H and MTF-1L. Zn reversibly modulated MTF-1H binding in a temperature-dependent manner. Similarly, Zn reversibly modulated MTF-1L binding, but, in contrast, such modulation occurred readily at 4 degrees C. This data demonstrate the conservation of binding specificity, binding properties, and regulation of MTF-1 in fishes. PMID- 11007175 TI - Properties of lactate dehydrogenase from the isopod, Saduria entomon. AB - Saduria entomon lactate dehydrogenase (LDH-A4*) from thorax muscle was purified about 89 fold to specific activity 510 micromol NADH/min/mg using Cibacron Blue 3GA Agarose and Oxamate-Agarose chromatographies. The enzyme is a tetramer, with molecular weight of 140 kDa for the native enzyme and 36 kDa for the subunit. The isoelectric point was at pH 5.7. The enzyme possesses high heat stability (T50 = 71.5 degrees C). The optimum pH for pyruvate reduction reaction was 6.5, while for lactate oxidation one, the maximum activity was at pH 9.1. The Km for pyruvate was minimal at 5 degrees C, the average environmental temperature of the isopod. The Km values determined at 30 degrees C and optimal pH for pyruvate reduction and lactate oxidation were 0.18 and 90.04 mM, respectively. Amino acid compositional analyses showed the strongest resemblance of the isopod isoenzyme to cod (Gadus morhua) LDH-C4. PMID- 11007176 TI - Astaxanthin is the major carotenoid in tissues of white storks (Ciconia ciconia) feeding on introduced crayfish (Procambarus clarkii). AB - We studied the carotenoid pigments in plasma, skin and body fat of white storks (Ciconia ciconia) from a colony in Spain feeding mainly on the recently introduced red swamp crayfish (Procambarus clarkii). In control colonies, where crayfish was absent, plasma was collected for comparison. Our objective was to determine whether the astaxanthin contained in the crayfish reached the blood, accumulated in fat, and finally was deposited in the red-colored bill and legs. If that was true, the visual cues provided by those tegumentary areas would be altered, with potential behavioral consequences. Plasma carotenoids were directly extracted with acetone, whereas skin and fat samples needed harsher conditions, i.e. grinding, sonication and extraction with diethyl ether. The extracts were analyzed by thin-layer chromatography (TLC) and UV/Vis spectroscopy. In crayfish eating storks, astaxanthin was confirmed to be the dominant pigment in all analyzed tissues. This red pigment was absorbed unchanged in the gut, and was responsible for the red color of plasma and the abnormal orange pigmentation of the feather-covered skin. It was also present in large quantities in the exposed bill and tarsi, which are typically red-colored in the stork. Control storks with no crayfish in the diet only presented lutein in their plasma. PMID- 11007177 TI - Effects of rabbit gastrointestinal mucins and dextran on hydrochloride diffusion in vitro. AB - We compared a viscous fingering formation of hydrochloric acid (HCl) in rabbit corpus, antral and duodenal mucins and with dextran under neutral and acidic conditions with respect to relative viscosity, molecular mass, and carbohydrate composition. The effect of desialyzation of duodenal mucin on the viscous fingering formation of HCl was also examined. HCl (0.1 N) was injected into 1% solutions of mucins and dextran and a subsequent viscous fingering formation was assessed based on an influx volume rate of HCl. A low influx volume rate indicates a high ability of the solutions to produce viscous fingers. The influx volume rate of HCl was lowest in duodenal mucin followed bl corpus mucin, antral mucin, and dextran at pH 7. The influx volume rate of HCl was inversely correlated with the relative viscosity of the solution. Maximum molecular masses were large in the order of corpus, antral, and duodenal mucins, and they were larger than dextran T2000. Rabbit gastrointestinal mucins were very polydisperse system. Duodenal mucin contains more sialic acid than gastric mucins; the influx volume rate of HCl increased in desialylated duodenal mucin. It is suggested that the higher ability of gastric mucins to prevent HCl diffusion than dextran were due to the differences in the molecular mass. The ability of duodenal mucin to prevent HCl diffusion was probably attributed to its high sialic acid content, which may reflect a physiological role of duodenal mucin in the duodenum that has to deal with HCl influx from the stomach. PMID- 11007178 TI - Oxygen transport proteins: III. Structural studies of the scorpion (Buthus sindicus) hemocyanin, partial primary structure of its subunit Bsin1. AB - The hemocyanin (Hc) from Buthus sindicus, studied in the native state, demonstrated to be an aggregate of eight different types of subunits arranged in four cubic hexamers. Both, the 'top' and the 'side' views of the native molecule have been identified from the negatively stained specimens using transmission electron microscopy. Out of these, eight different polypeptide chains, the partial primary structure (68%) of a subunit Bsin1 (Mr = 72422.7 Da) was established using a combination of automated Edman degradation and mass spectrometry. A multiple sequence alignment with other closely related cheliceratan Hc subunits revealed average identities of ca. 60%. Most of the structurally important residues, i.e. copper and calcium-binding ligands, as well as the residues involved in the presumed oxygen entrance pathway, proved to be strictly conserved in Bsin1. Sequence variations have been observed around the functionally important chloride-binding site, not only for the B. sindicus subunit Bsin1, but also for the subunit Aaus-6 of the scorpion A. australis and the subunit Ecal-a from the spider Eurypelma californicum Hcs. Deviation in the primary structure related to the chloride-binding site suggest that the effect of chloride ions may vary in different hemocyanins. Furthermore, the secondary structural contents of the Hc subunit Bsin1 were determined by circular dichroism revealing ca. 33% alpha-helix, 18%, beta-sheet, 19% beta-turn, and 30% random coil composition. These values are in good agreement with the crystal structure of the closely related Hc subunit Lpol-II from horseshoe crab L. polyphemus. Electron microscopic studies of the purified Hc subunit under native conditions revealed that Bsin1 has self aggregation properties. Results of these studies are discussed. PMID- 11007179 TI - Cross-reactivities of polyclonal antibodies against factor V activating enzyme, a serine proteinase from Vipera lebetina (snake) venom. AB - Antibodies to the factor V activating enzyme from Vipera lebetina venom were produced by immunizing a rabbit with chromatographically purified factor V activating enzyme probes. The antibodies cross-reacted with different protein fractions in 23 snake venoms (ten viperid, eight crotalid, and five elapid venoms) as demonstrated by western immunoblotting. In the venom of Vipera russelli the antibodies recognized only one protein band which probably belonged to factor V activating enzyme. PMID- 11007180 TI - In vitro polymerization of mussel polyphenolic proteins catalyzed by mushroom tyrosinase. AB - The in vitro enzymatic polymerization of the polyphenolic protein purified from the mussels Aulacomya ater, Mytilus edulis chilensis and Choromytilus chorus was studied. Mushroom tyrosinase was used to oxidize the dopa residues present in these proteins, and polymerization was monitored by acid-urea polyacrylamide gel electrophoresis. The protein from A. ater polymerized at a faster rate than the other two. Amino acid analysis of the crosslinked protein showed a notable decrease in the content of dopa, but no significant change of other amino acids. This suggests that crosslink formation may be limited to the oxidized dopa derivatives of the protein molecules. PMID- 11007181 TI - Metabolites from a marine bacterium Pseudomonas/Alteromonas, associated with the sponge Dysidea fragilis. AB - The lipophylic extract from a Black Sea bacterium, associated with the sponge Dysidea fragilis, was investigated. Saturated hydrocarbons and fatty acids of the lipids were identified. The concentrations of the polycyclic compounds appeared to be negligible. The main components appeared to be phosphatidyl ethanolamine, followed by phosphatidyl serine. The first was investigated by FAB mass spectrometry and a series of molecular species partially identified. PMID- 11007182 TI - Partial characterization of vitellin and localization of vitellogenin production in the terrestrial isopod, Armadillidium vulgare. AB - Vitellins were purified separately from ovaries and eggs of the isopod, Armadillidium vulgare. Ovarian vitellin consisted of at least six proteins with relative molecular masses of 205, 200, 185, 180, 122 and 112 kDa. The larger four proteins disappeared in eggs within a week after oviposition and a 59-kDa protein appeared thereafter. The amino-terminal amino acid sequences of these vitellin proteins were identical except for the ovarian 112 kDa, egg 112 kDa and 59 kDa proteins, and showed considerable similarity to those of known vitellogenins from other animals. Comparison of tryptic peptide maps of the 122 and 112 kDa proteins from eggs on reversed-phase HPLC and sequence identification of two randomly selected peaks having the same retention times indicated that two peptides were mostly similar in sequence. PCR-assisted cloning of the 5' region of a cDNA (591 bp) encoding vitellogenin revealed the presence of a signal peptide consisting of 16 amino acid residues and clarified the structural relationship among the protein components except for the ovarian 112 kDa and the egg 59 kDa proteins. Northern blot analysis revealed that the fat body is the main vitellogenin producing organ. PMID- 11007183 TI - Succinate and alanine as anaerobic end-products in the diving turtle (Chrysemys picta bellii). AB - The western painted turtle is an extremely anoxia-tolerant vertebrate capable of tolerating blood lactate levels of 150-200 mM. Since lactate increases to such high levels, other fermentation end-products such as succinate and alanine, which have not been previously measured in this species, might also be expected to increase. Therefore, I measured turtle heart, liver, and blood concentrations of lactate, succinate, and alanine following a 28-day anoxic dive at 5 degrees C. Succinate and lactate concentrations increased significantly in all three compartments while alanine increased significantly in the liver only. Lactate was found to accumulate by a similar amount in all three compartments (66.4-80.5 micromol g or ml(-1) in the blood compartment) and was used as a reference to which alanine and succinate concentrations could be compared. Succinate and alanine levels increased by 2 and 0.9% of lactate in liver, approximately 0.3 and 0.04% of lactate in blood, and 0.6 and 0.07% of lactate in heart, respectively. The contribution of each to the total anoxic heat production was calculated and accounted for an additional 1.5% of the previously measured exothermic gap. I conclude that succinate and alanine concentrations do increase in the anoxic turtle but are minor anaerobic end-products. PMID- 11007184 TI - Differential expression of sialic acid on porcine organs during the maturation process. AB - Sialylated structures play important roles in cell communication, and change in a regulated manner during development and differentiation. In this work, we report the main glycosidic modifications that occur during the maturation of porcine tissues, involving the sialylation process as determined with lectins. Sialic acids were identified at several levels in a broad range of cell types of nervous, respiratory, genitourinary and lymphoid origin. Nevertheless, the most contrasting was the type of glycosidic linkage between 5-N-acetyl-neuraminic acid (Neu5Ac) and galactose (Gal) expressed in central nervous system (CNS). Newborn CNS abundantly expressed Neu5Acalpha2,3Gal, but weakly or scarcely expressed Neu5Acalpha2,6Gal/GalNAc. Maturation of CNS induced drastic changes in sialic acid expression. These changes include decrease or complete loss of NeuAcalpha2,3Gal residues, mainly in olfactory structures and brain cortex, which were replaced by their isomers Neu5Acalpha2,6Gal/GalNAc. In the brain cortex and cerebellum, the increase of Neu5Acalpha2,6Gal/GalNAc molecules was paralleled by an increase of 5-N-acetyl-9-O-acetyl-neuraminic acid (Neu5,9Ac2). In addition, terminal Gal and N-acetyl-D-galactosamine (GalNAc) residues also increased their expression in adult CNS tissues, but this was more significant in structures forming the encephalic trunk. Our results show that sialylation of porcine CNS is finely modulated throughout the maturation process. PMID- 11007185 TI - Digestive amylase from the larger grain borer, Prostephanus truncatus Horn. AB - A combination of ion-exchange chromatography, preparative electrophoresis and gel filtration chromatography allowed a 1209-fold purification of one of the two major digestive alpha-amylases from larvae of the larger grain borer, Prostephanus truncatus Horn. The purified enzyme showed a molecular mass of 60.2 kDa, an isoelectric point of 4.7 and an optimal pH for activity of 6.0. The enzyme was heat labile and it was recognized by proteinaceous inhibitors from amaranth seeds (Amaranthus hypochondriacus), whereas extracts from maize (Zea mays) and tepary bean (Phaseolus acutifolius) produced very low inhibition. When the enzyme was measured at different stages of development, maximal activity was found in the second instar larvae. Activity drastically decreased to a very low level during the pupae stage and increased again at the adult stage. A zymogram of the different developmental stages showed two main bands of alpha-amylase activity, which almost disappeared at the pupae stage to increase again during the adult stage, revealing a new, smaller band. This new band may be required for a better adaptation of the adult insect to its new environment. PMID- 11007186 TI - Gene products from LUQ neurons in the abdominal ganglion are present at the renal pore of Aplysia californica. AB - The L2-4,6 and L5 cells located in the left upper quadrant of the abdominal ganglion of Aplysia californica express the L5-67 and LUQ-1 genes, respectively, in a nonoverlapping manner. These cells send major neurites to the kidney and at least some of them were shown to innervate the renal pore closer muscle, and thereby control its function. By using in-situ hybridization and immunofluorescence, the presence of L5-67 and LUQ-1 mRNAs and peptides was studied in the kidney, with emphasis on the region of the renal pore. We detected immunoreactive materials in many small varicose nerve fibers running along the central epithelium in the inner parts of the kidney, and in neurites located within a large nerve associated with muscles inside the renal pore. Our observations represent the first direct evidence of the presence of gene products from LUQ cells at the renal pore, suggesting that they may be responsible for mediating LUQ cell signals. Furthermore, mRNAs coding for the L5-67 and LUQ-1 peptides were also found in the nerve structure inside the renal pore. Our report documents a striking example of neuropeptide mRNA targeting nerve terminals that are very distant from their cell bodies. PMID- 11007187 TI - Phosphatidylcholine-specific phospholipase C-mediated induction of phospholipase D activity in Fas-expressing murine cells. AB - We have previously reported that Fas cross-linking resulted in the activation of phosphatidylcholine-specific phospholipase C (PC-PLC) and the subsequent activation of protein kinase C (PKC) and phospholipase D (PLD) in A20 cells. In an attempt to correlate the existence of PC-PLC activity and activation of PLD by Fas activation among various Fas-expressing murine cell lines, we have investigated the effect of anti-Fas monoclonal antibody on PC-PLC and PLD activities in A20, P388D1 and YAC-1 cell lines. Upon treatment of anti-Fas monoclonal antibody to these three cell lines, the activation of PLD was only observed in A20 cells. When the effect of anti-Fas monoclonal antibody on PKC and PC-PLC activities in Fas-expressing clones were investigated, the activation of PKC and PC-PLC was detected only in A20 clones. Results presented here also show that exogenous addition of Bacillus cereus PC-PLC activates PC hydrolysis, PKC and PLD in all three murine cell lines. These findings suggest that the activation of PC-PLC is a necessary requirement for the activation of PLD by Fas cross-linking and cell lines devoid of functional PC-PLC activity could exhibit enhanced PLD activity by exogenous addition of PC-PLC. PMID- 11007188 TI - Introduction to the Margaux Conference on Critical Illness: activation of the coagulation system in critical illnesses. PMID- 11007189 TI - Pathophysiologic basis of sepsis: considerations for future strategies of intervention. AB - OBJECTIVE: To analyze the implications for therapeutic advances in the treatment of sepsis and septic shock based on the mechanisms underlying the response to microbial infection. DATA SOURCES: Clinical trials and experimental models designed to determine the key mechanisms involved in cellular and molecular processes of inflammatory reactions. STUDY SELECTION: Analyses of normal immune reactions to microbial infection, processes involved in the development of sepsis, and reasons for frequent failure of regimens based on current therapeutic rationales. DATA EXTRACTION/SYNTHESIS: Review of the data suggests that the dysregulation of the immune system resulting in sepsis/septic shock is ineffectually blocked by interfering with the action of inflammatory mediators or cascades, as these processes may be too complex to be easily antagonized. Rather, blockade of the action of microbial products or of the intracellular processes activated by receptor interaction with these products may provide a more promising therapeutic alternative. CONCLUSIONS: The sepsis syndrome induced by microbial pathogens reflects excessive stimulation of the processes of innate immunity. Bacterial components reacting with specific receptors activate intracellular processes, resulting in the release of excessive amounts of mediators of inflammation as well as triggering multiple complex proteolytic cascades. Blockade or antagonism of the actions of individual intermediary messenger molecules has proved therapeutically unsuccessful, because blockade of mediators alone does not block the direct activation of processes such as coagulation and complement. The dysregulation that characterizes sepsis may be amenable to blockade of the bacterial components or to the intracellular pathways triggered by these products. PMID- 11007190 TI - Pathophysiology of disseminated intravascular coagulation in sepsis. AB - Disseminated intravascular coagulation (DIC) is an acquired syndrome characterized by intravascular fibrin formation occurring in the course of a variety of severe diseases. In gram-negative sepsis, endotoxin is the bacterial component eliciting a cascade of tissue factor dependent hypercoagulable reactions mediated by cytokines, including tumor necrosis factor-alpha and interleukin-6. Fibrinolysis is activated in this process by the action of tumor necrosis factor-alpha, but its activity is impaired by the predominant inhibitory effect of plasminogen activator inhibitor-1. Natural inhibitory mechanisms include antithrombin, the protein C system, and tissue factor pathway inhibitor. Each of these defense systems counteracts the harmful effects of DIC, and its acquired deficiency is associated with increased mortality in observational studies. The generation of several proteases in DIC, including factor Xa and thrombin, has potential interactions with inflammatory pathways that may potentiate the systemic inflammatory syndrome that often accompanies DIC. Experimental studies support the notion that defects in the protein C pathway modulate the inflammatory response, and illustrate that coagulation and inflammation are coupled systems in DIC. PMID- 11007192 TI - Novel approaches to the management of disseminated intravascular coagulation. AB - OBJECTIVE: Disseminated intravascular coagulation (DIC) is a syndrome characterized by systemic intravascular activation of coagulation, leading to widespread deposition of fibrin in the circulation. We addressed the issue of whether there is evidence that this fibrin deposition contributes to multiple organ failure. We also explored the current knowledge on the pathogenesis of DIC and reviewed current and future treatment for DIC. DATA SOURCES: We searched and reviewed published articles on experimental studies of DIC models in animals and clinical studies in patients with DIC. DATA SYNTHESIS: There is ample experimental and clinical evidence that DIC contributes to morbidity and mortality. Recent knowledge on important pathogenetic mechanisms that may lead to DIC has resulted in novel preventive and therapeutic approaches to patients with DIC. Although the trigger for the activation of the coagulation system may vary depending on the underlying condition, it is usually mediated by several cytokines. Thrombin generation proceeds via the (extrinsic) tissue factor/factor VIIa route and simultaneously occurring depression of inhibitory mechanisms, such as antithrombin III and the protein C-protein S system. Also, impaired fibrin degradation, because of high circulating levels of plasminogen activator inhibitor, type 1, contributes to enhanced intravascular fibrin deposition. CONCLUSIONS: Although the cornerstone of DIC management is the specific and vigorous treatment of the underlying disorder, strategies aimed at inhibiting coagulation activation may theoretically be justified. Such strategies have been found to be beneficial in experimental and initial clinical studies. These strategies, which follow from our current understanding of the pathophysiology of DIC, involve inhibition of tissue factor-mediated activation of coagulation or restoration of physiologic anticoagulant pathways by means of the administration of antithrombin concentrate or (activated) protein C concentrate. Although no complete evidence from controlled clinical trials is available for most of the proposed therapeutic interventions, these novel strategies are being studied. PMID- 11007191 TI - Description of compensated and uncompensated disseminated intravascular coagulation (DIC) responses (non-overt and overt DIC) in baboon models of intravenous and intraperitoneal Escherichia coli sepsis and in the human model of endotoxemia: toward a better definition of DIC. AB - OBJECTIVE: Work toward a better definition of disseminated intravascular coagulation (DIC) by characterizing the difference between compensated and uncompensated responses of the hemostatic system to inflammatory stress in baboons and human subjects using global coagulation and molecular marker assays of hemostatic, inflammatory, and endothelial perturbation. DESIGN: We conducted prospective evaluation of the response of baboons to increasing concentrations of intravenous Escherichia coli, human subjects to intravenous endotoxin, and baboons to intraperitoneal E. coli. SETTING: Animal laboratory and medical intensive care facilities, University of Oklahoma Medical School laboratories. SUBJECTS: Cynocephalus baboons; normal healthy male human subjects (age, 24-37 yrs). MEASUREMENTS AND MAIN RESULTS: Global coagulation assays, white blood cell counts, and molecular marker assays (ELISA) of components of the inflammatory and hemostatic systems, neutrophil release products, and endothelial injury. A fall in both fibrinogen concentration and platelet counts indicated a decompensated hemostatic response to inflammatory stress (ie, overt DIC). These responses were observed 2-6 hrs after intravenous infusion of 10(9) and 10(10) colony-forming units (CFU)/g of E. coli and after implantation of 10(11) CFU/g of E. coli into the peritoneal cavity. However, 6 hrs after E. coli challenge, these tests were much less reliable as markers of overt DIC because the fibrinogen underwent an acute phase response and the platelet count fell and remained depressed for 48 hrs in the face of a coagulopathic response that was already beginning to resolve, as reflected by a rising fibrinogen concentration. This lack of reliability was particularly evident in the E. coli peritonitis studies, in which one third of the animals recovered, one third remained sick for up to 14 days, and one third died. In contrast, fibrin degradation products and the molecular markers thrombin/antithrombin, soluble fibrin monomer, protein C, and activated protein C/inhibitor complexes responded consistently in a dose-dependent manner regardless of the length of time after challenge. These variables exhibited this dose response to 106 and 108 CFU/g of E. coli in absence of a fall in fibrinogen concentration. This was defined as a compensated hemostatic response to inflammatory stress (ie, non-overt DIC). The values of these variables correlated closely with rising concentrations of markers of neutrophil activation (elastase/alpha 1 antitrypsin) and endothelial injury (soluble thrombomodulin). This was particularly evident in the human response to endotoxin, in which there was abundant evidence of hemostatic marker response in absence of a fall in platelet or fibrinogen concentration, both immediately after endotoxin infusion (first stage, 0-8 hrs after endotoxin) and later (second stage, 12-48 hrs after endotoxin). CONCLUSION: Global coagulation tests are most useful in detecting overt consumptive coagulopathy (overt DIC) near the time of challenge or injury (1 to 6 hrs). Molecular markers can detect and grade the degree of hemostatic stress of a non-overt consumptive coagulopathy (nonovert DIC). These markers correlate with degree of endothelial cell injury and reveal a reperfusion injury stage (second stage) in the human endotoxin model of compensated hemostatic stress after all clinical symptoms have subsided and the subjects have returned to work. PMID- 11007193 TI - Tissue factor pathway of coagulation in sepsis. AB - OBJECTIVE: To review the role of the tissue factor pathway of coagulation in experimental sepsis. DATA SOURCES: Studies published in biomedical journals. STUDY SELECTION: Studies on the role of the tissue factor pathway in animal or human models for sepsis. DATA EXTRACTION AND SYNTHESIS: Variables reflecting tissue factor pathway activation in the various models are discussed; the effects of administration of tissue factor pathway inhibitors on these and inflammatory variables, as well as on the course and outcome, are analyzed. CONCLUSION: Activation of coagulation during experimental sepsis occurs mainly, if not exclusively, via the tissue factor pathway; inhibitors of this pathway improve mortality, presumably by a combined attenuating effect on coagulative and inflammatory responses. PMID- 11007194 TI - Tissue factor inhibition and clinical trial results of tissue factor pathway inhibitor in sepsis. AB - Tissue factor mediated pathways leading to microvascular thromboses and endothelial activation appear to play an important role in the development of multiple organ failure associated with severe sepsis. Tissue factor pathway inhibitor (TFPI) is an endogenous inhibitor of tissue factor associated coagulation cascades. In experimental models of severe sepsis, treatment with TFPI results in significant reduction in mortality. Similarly, a recently completed Phase II 210-patient study comparing placebo and infusions of TFPI showed trends toward a relative reduction in day 28 all-cause mortality in TFPI treated patients. These data suggest that coagulation cascades involving tissue factor contribute to organ dysfunction in critically ill septic patients. TFPI may be a useful therapy in improving outcome of severe sepsis. PMID- 11007195 TI - Therapeutic rationale for antithrombin III in sepsis. AB - OBJECTIVE: To review the preclinical evidence that provides the therapeutic rationale for antithrombin as a novel treatment for human sepsis. DATA SOURCES: A summary of published medical literature from MEDLINE search files and other reviews published about antithrombin use in sepsis. DATA SUMMARY: Antithrombin has a variety of antiinflammatory properties in addition to its functions as an endogenous anticoagulant that appear to have an important therapeutic role in the prevention of microvascular dysfunction and multiple organ injury in sepsis. Appropriate timing and dosing of antithrombin III is critical to realize its full therapeutic potential as an anti-sepsis therapy. CONCLUSIONS: Antithrombin is a potent inhibitor of thrombin-mediated vascular injury in the microcirculation in severe sepsis. This endogenous anticoagulant is rapidly depleted in the early phases of sepsis as a result of decreased synthesis, increased destruction, and enhanced clearance by thrombin-antithrombin complex formation. The therapeutic efficacy of antithrombin in experimental sepsis is readily demonstrable in numerous animal systems. Appropriately defined patient populations with early onset severe sepsis and/or septic shock may benefit from antithrombin therapy if it is administered in adequate doses at the optimal time interval. PMID- 11007196 TI - Clinical trial results with antithrombin III in sepsis. AB - OBJECTIVE: To present and discuss the rationale and results of clinical trials using antithrombin (AT) supplementation in patients with sepsis. DATA SOURCES/STUDY SELECTION: Review of all controlled (open or double-blind) studies of patients with severe sepsis or septic shock who were treated with AT concentrates to obtain better control of coagulation activation and inflammation. DATA EXTRACTION: AT is a major inhibitor of the coagulation cascade. Recent experimental studies have also shown that it can modulate the inflammatory reactions that occur during sepsis. An early and prolonged decrease in AT activity is well documented during sepsis-induced disseminated intravascular coagulation and during the systemic inflammatory response. Thus, supplementation with AT concentrates has been proposed as a potential therapy in sepsis patients. DATA SYNTHESIS: Numerous uncontrolled studies of AT supplementation in sepsis patients have been reported in the last 20 yrs. Since 1993, four placebo controlled randomized studies have been performed in France, Germany, Northwestern Europe, and Italy. Three of these studies were subjected to a meta analysis of 122 patients. Results showed a nonsignificant 22% reduction in the 30 day all-cause mortality and a reduction in the length of stay in the intensive care unit in the AT treated group. The Italian study of 120 patients demonstrated that the overall mortality was similar in the placebo and treated groups. However, post hoc analysis according to the Cox regression model showed that in patients with septic shock, AT supplementation significantly decreased the risk of death. CONCLUSIONS: Together, these studies are consistent with the positive effect seen with AT supplementation in patients with severe sepsis. A multicenter phase III trial is currently in progress to definitively document its effect on mortality. PMID- 11007197 TI - The protein C pathway. AB - OBJECTIVE: To examine the role of the protein C anticoagulant pathway in the regulation of microvascular thrombosis. The mechanisms by which inflammation impairs the function of this pathway are also reviewed; conversely, we will survey emerging knowledge of the multiple mechanisms by which the protein C anticoagulant pathway can control the inflammatory response. DATA SOURCES: The information reviewed here was taken from the primary literature, including recent abstracts. STUDY SELECTION: All studies that bear directly on the interrelationship between the protein C anticoagulant pathway and inflammation were included, as was a summary of the initial clinical experience with protein C/activated protein C therapy in sepsis. DATA EXTRACTION AND SYNTHESIS: The results from each of the experimental approaches are summarized. Clinical experience with protein C supplementation in sepsis, although promising, is still in the early stages of study. CONCLUSIONS: The protein C anticoagulant pathway is a major mechanism in controlling microvascular thrombosis. Protein C deficiency that can occur in sepsis facilitates thrombin generation in the microvasculature, probably augmenting inflammatory responses and contributing to endothelial cell dysfunction. Animal studies and preliminary clinical results suggest that protein C/activated protein C supplementation may be useful in reversing microvascular dysfunction. PMID- 11007198 TI - Protein C levels as a prognostic indicator of outcome in sepsis and related diseases. AB - OBJECTIVE: To consider the appropriateness of protein C levels as a prognostic indicator for sepsis and related diseases. DATA SOURCES/STUDY SELECTION: Published research and review articles related to protein C deficiency in patients with sepsis and related diseases. DATA EXTRACTION AND SYNTHESIS: All applicable data were extracted, and relevant literature was cited to support factual statements in the text. The protein C pathway represents one of the major regulatory systems of hemostasis, exhibiting antithrombotic, profibrinolytic, and anti-inflammatory properties. Numerous studies have shown that acquired protein C deficiency is prevalent in the majority of septic patients (>85%) and is associated with increased morbidity and mortality in patients with severe sepsis and septic shock. This deficiency in protein C is not simply a transient marker for sepsis, but parallels the progress of the disease. In addition, protein C deficiency occurs in the presence of a wide range of pathogens and develops early in the disease process. CONCLUSIONS: A review of the relevant literature suggests that protein C levels may serve as a useful prognostic indicator of outcome in sepsis and related diseases. PMID- 11007199 TI - Meningococcemia as a model for testing the hypothesis of antisepsis therapies. AB - OBJECTIVE: To critically review the advantages and disadvantages of pediatric meningococcemia as a model for testing antisepsis therapies. DATA SOURCES: Research and review articles on the pathogenesis and treatment of human meningococcemia, as well as editorial commentaries discussing the failure of clinical trials for adult sepsis or Systemic Inflammatory Response Syndrome. Data from these sources are presented in the context of the author's experience as principal investigator in a large, randomized trial on children with invasive meningococcal disease. STUDY SELECTION AND DATA EXTRACTION: Studies were selected to include aspects of epidemiology, pathophysiology, outcome prediction, and therapeutic trials. DATA SYNTHESIS: Compared with an adult sepsis population, meningococcemia is a single disease, diagnosed clinically with high reliability. Patients are previously healthy, without underlying medical or surgical conditions. In contrast to sepsis trials, nearly all patients with meningococcal disease receive effective antibiotics. Finally, meningococcemia most closely resembles animal models of endotoxin infusion, in which most antisepsis therapies have been highly effective. However, the meningococcal model carries major disadvantages, among them that meningococcemia is rare and rapidly progressive and patients are widely dispersed geographically. In addition, a wide range of experimental therapies is routinely provided in an attempt to preserve life or limbs. CONCLUSIONS: Meningococcemia is an ideal model of a rapidly progressive bacterial infection associated with marked endotoxemia. Problems with the model can be overcome by extensive pretrial logistic planning, as well as close coordination and cooperation with national regulatory agencies. PMID- 11007200 TI - Do new strategies in meningococcemia produce better outcomes? AB - Meningococcal septic shock (MSS) has high mortality and morbidity rates. In addition to the traditional prompt antibiotics and respiratory and circulatory support, new treatment strategies have been proposed. AGAINST THE INFLAMMATORY CASCADE: Immunotherapy, such as antiserum to Escherichia coli J5 and human antilipid A monoclonal antibodies/centoxin (HA-1A), did not significantly alter the mortality rate of MSS; we are awaiting the results of the bactericidal/permeability-increasing protein multicenter trial. Two series reported the effects of hemofiltration and hemodiafiltration in MSS, but the true benefits remain unknown. TO TREAT HEMOSTATIC ABNORMALITIES: In MSS, heparin is still controversial and antithrombin concentrate use has been reported in only one child. Several case reports on protein C and recombinant tissue plasminogen activator have been published; the efficacy (improvement in shock and organ dysfunction and reduction in amputation rate) and safety (intracerebral hemorrhage with recombinant tissue plasminogen activator) of these treatments need further evaluation. Blood and plasma exchange appear to be safe and are supposed to reduce mortality, but it is difficult to draw firm conclusions from published studies. Finally, local application of medicinal leeches has been reported to improve purpuric lesions. TO INDUCE VASODILATION: Prostacyclin, or epoprostenol, infusion, sodium nitroprussiate infusion, sympathetic blockade, and topical nitroglycerin have been reported to improve distal perfusion; however, these reports are all anecdotal. OTHER STRATEGIES: Improvement in limb perfusion was achieved after hyperbaric oxygenation in patients with purpura fulminans caused by different pathogens. Most authors recommend monitoring of compartment pressures and performing fasciotomy as indicated. Finally, extracorporeal membrane oxygenation was recently proposed to support seven children with intractable MSS. CONCLUSIONS: There is no proof that unconventional treatments have a significant impact on outcome in MSS; prospective multicenter trials are needed. At present, early recognition of meningococcal sepsis and appropriate treatment seem to be the optimal methods of improving outcome. Efforts to find an effective meningococcal vaccine must be continued. PMID- 11007201 TI - Meningococcal disease as a model to evaluate novel anti-sepsis strategies. AB - OBJECTIVE: To consider the appropriateness of meningococcemia as a clinical entity for the evaluation of anti-endotoxin agents. DATA SOURCES: English language published literature concerning meningococcemia, sepsis, and endotoxin. STUDY SELECTION: The purpose of this work is to consider some of the practical and scientific issues that arise in designing clinical trials to evaluate anti endotoxin agents. A selected review of recently published articles was undertaken. DATA EXTRACTION AND SYNTHESIS: Relevant literature has been cited to support factual statements in the text. CONCLUSION: Meningococcemia as a paradigm of endotoxin-mediated Gram-negative sepsis has many advantages. It is a homogeneous population, and it represents a single, measurable, bacterial target that is unequivocally implicated in the pathogenesis of the disease. However, it is an uncommon disease that may develop so quickly that attempts to intervene may be too late to have an effect. There is considerable morbidity, but how best to measure the outcome and the extent to which the results can be extrapolated to adult populations with sepsis secondary to nosocomial infection remain unclear. PMID- 11007202 TI - Coagulation inhibitor replacement in sepsis is a potentially useful clinical approach. AB - In sepsis, levels of the endogenous coagulation inhibitors antithrombin III and protein C are lowered as a result of complex formation with multiple activated clotting factors. In addition, their activity can further be curtailed by proteolytic inactivation. Loss of antithrombin III and protein C activity blocks the endogenous control mechanism for thrombin generation resulting in a state of systemic activation of coagulation and inflammatory processes. Levels of tissue factor pathway inhibitor, a third endogenous coagulation inhibitor, are increased in sepsis rather than decreased, probably reflecting a depletion of the endothelial cell bound tissue factor pathway inhibitor pool with loss of its endothelial protective function. Administration of any of these three inhibitors in various animal species and sepsis models reduces morbidity and mortality. In addition to their anticoagulant effects, these inhibitors also have various anti inflammatory activities that may contribute to their protective effects. Phase II studies in patients with severe sepsis using coagulation inhibitors have indicated that this therapeutic approach may be useful. Large-scale phase III trials will ultimately decide whether adjunctive coagulation inhibitor replacement will have a place in the treatment of patients with severe sepsis. PMID- 11007203 TI - Coagulation inhibitor replacement during sepsis: useless? AB - OBJECTIVE: Because coagulatory activation in sepsis is triggered mainly by tissue factor release from endothelial cells and blood monocytes during their activation via proinflammatory cytokines, inhibition of coagulation by exogenous administration of coagulation inhibitors has been proposed. These strategies should allow us to prevent and treat excessive coagulatory activation, thereby potentially preventing sepsis-induced organ dysfunction. Potential therapies include the natural coagulation inhibitors antithrombin, activated protein C, and tissue factor pathway inhibitor, as well as direct thrombin inhibition by recombinant hirudin. DATA SOURCES: A limited review of the published literature using all sources was undertaken. STUDY SELECTION: Selected clinical and experimental studies with coagulatory inhibitors were analyzed. CONCLUSIONS: The biological properties of coagulatory activation during sepsis (coagulation as a protective mechanism to control the septic focus, e.g., fibrin deposition during peritonitis) are not completely understood. Therefore, one has to be careful when administering coagulatory inhibitors, especially because patients with multiple organ dysfunction syndrome often do not show the widespread fibrin deposition in nutritive blood vessels that have been seen experimentally. How might these patients benefit from thrombin inhibition? Coagulatory activation per se seems unlikely to directly cause deterioration of organ function, although it is involved in generalized endothelial activation with consecutive mediator release and increased leukocyte-endothelial cell interaction. Antagonism of inflammatory mediators and, consecutively, endothelial cell activation might be a better target in adjunctive sepsis therapy, with improvement in septic microcirculatory disturbances. Administration of natural pleiotropic coagulation inhibitors that are documented positive effects on the microcirculation, (such as activated protein C, antithrombin) seems to be promising. PMID- 11007204 TI - Phylogenetic and functional relationships between coagulation and the innate immune response. AB - Activation of the coagulation system frequently accompanies systemic inflammatory states and is an almost invariable consequence of septic shock. The simultaneous activation of the innate immune response and the coagulation system after injury is a phylogenetically ancient, adaptive response that can be traced back to the early stages of eukaryotic evolution. Most invertebrate species lack differentiated phagocytic cells and platelets. They possess a common cellular and humoral pathway of inflammation and clotting after a breach in their internal milieu by either trauma or infection. The close linkage between clotting and inflammation has been preserved throughout vertebrate evolution and is readily demonstrable in human physiologic responses to a variety of potentially injurious stimuli. The same proinflammatory stimuli that activate the human clotting cascade also activate the phagocytic effector cells (neutrophils, monocytes, and macrophages). The complex and highly integrated linkage between systemic inflammation and coagulation are reviewed in this article. PMID- 11007205 TI - Roundtable I: relationships between coagulation and inflammatory processes. PMID- 11007206 TI - New therapeutic implications of anticoagulation mediator replacement in sepsis and acute respiratory distress syndrome. AB - OBJECTIVE: To examine the relationship between the coagulation and immune systems in sepsis and acute respiratory distress syndrome and to review published experimental and clinical studies that use anticoagulation mediator replacement strategies in these conditions. DATA SOURCES: MEDLINE database and bibliographies of relevant articles. STUDY SELECTION: Articles (both original and review) relating to coagulation abnormalities and anticoagulation mediator replacement therapy in sepsis and acute respiratory distress syndrome. DATA EXTRACTION: All applicable data were extracted. CONCLUSIONS: Coagulation abnormalities are common in the critically ill. Early studies using anticoagulation mediator replacement in patients with sepsis have suggested beneficial effects on organ function and outcome. The results from larger, randomized controlled trials are eagerly anticipated. PMID- 11007207 TI - Roundtable II: clinical implications of anticoagulation mediator replacement in sepsis and acute respiratory distress syndrome. PMID- 11007208 TI - Concluding discussion of the Margaux Conference on Critical Illness: activation of the coagulation system in critical illnesses. PMID- 11007209 TI - Esophagogastric cancer--time to change the paradigm. PMID- 11007210 TI - Autism and the gastrointestinal tract. PMID- 11007211 TI - The effect of alcohol on body composition. PMID- 11007212 TI - Management of recurrent hepatitis C after liver transplantation: a concise review. AB - Recurrent hepatitis C infection and subsequent graft failure are increasingly recognized problems after orthotopic liver transplantation. Although many prospective therapeutic, controlled trials in primary hepatitis C disease have been reported, large-scale studies are yet to be performed in patients with posttransplant recurrent hepatitis C after liver transplantation. In this review, we summarize the current literature on the therapeutic approaches for recurrent hepatitis C and discuss the results of published studies on therapy with ribavirin or interferon (IFN) alone and on combination therapy with IFN plus ribavirin. Further, we discuss results of prophylactic approaches to the problem of recurrent hepatitis C after transplant. Finally, we discuss additional aspects of anti-hepatitis C virus therapy after liver transplantation. PMID- 11007213 TI - Herpes simplex virus esophagitis in the immunocompetent host: an overview. AB - OBJECTIVE: The aim of this study was to delineate the characteristics of herpes simplex virus esophagitis (HSVE) in the immunocompetent host. METHODS: The study entailed a case report and a review of relevant literature through a MEDLINE search back to 1966. All cases with documented HSVE in patients without immunosuppression were selected and their characteristics defined. RESULTS: A total of 38 cases were identified. The age range was 1-76 yr and the male/female ratio 3.2/1. Antecedent exposure to HSV disease was described in eight cases (21.1%). A prodrome of systemic manifestations preceded the onset of esophageal symptoms in nine subjects (23.6%). Manifestations included acute odynophagia (76.3%), heartburn (50%), and fever (44.7%). Concurrent oropharyngeal lesions were uncommon (n = 8, 21.1%). Endoscopically, extensive involvement was common, showing friable mucosa (84.2%), numerous ulcers (86.8%), and whitish-exudates (39.5%). The distal esophagus was most commonly affected (63.8%). Microscopic examination showed characteristic viral cytopathology in 26 (68.4%) cases. Virus was recovered from esophageal-brushes or biopsies in 23 of 24 (95.8%) patients and immunocytochemistry was positive in seven of eight (87.5%) cases. Immune status was consistent with primary HSV infection in eight (21.1%) cases. The disease was self-limiting, although esophageal perforation and upper GI bleeding were reported in one case each. CONCLUSIONS: HSVE in the immunocompetent host is a rare but distinct entity, and is significantly more common in male subjects. It represents either primary infection or reactivation, and is characterized by acute onset, systemic manifestations, and extensive erosive-ulcerative involvement of the mid-distal esophagus. Histopathological examination alone may miss the diagnosis; adding tissue-viral culture optimizes the diagnostic sensitivity. It is usually self-limiting; whether antiviral therapy is beneficial remains unknown. PMID- 11007214 TI - Results of photodynamic therapy for ablation of dysplasia and early cancer in Barrett's esophagus and effect of oral steroids on stricture formation. AB - OBJECTIVE: The primary goal of this study was to investigate whether oral steroids would reduce the incidence of stricture formation after photodynamic therapy (PDT) in Barrett's patients. The effect of balloon window length, pretreatment of nodules, retreatment of skipped areas, and subsequent PDT on the incidence of strictures was also investigated. The ultimate goal of treatment was elimination of dysplasia, early cancer, and Barrett's mucosa. METHODS: A total of 60 patients were injected with Photofrin (2 mg/kg). Patients were randomized to receive PDT (n = 30) or PDT and oral prednisone (n = 30). Two days later, 630 nm light (KTP/dye laser) was delivered using a 5- or 7-cm windowed balloon at a light dose of 200 or 175 J/cm. The majority of patients received 200 J/cm using a 7-cm balloon. Nodules were pretreated with a short diffuser at a dose of 50-75 J/cm. Additional light was delivered to skipped areas 2-3 days later. Endoscopies were conducted every 3-6 months to evaluate the response. Residual or recurrent Barrett's was treated using neodymium:aluminum-yttrium-garnet (Nd:YAG) laser (small areas) or was retreated with PDT. RESULTS: The effect of steroids on the incidence of strictures was analyzed in patients receiving a single treatment with a light dose of 200 J/cm using a 7- cm balloon. There was no reduction in the incidence of strictures in patients receiving PDT and steroids (29%) compared to those receiving PDT alone (16%). Treatment using a 7-cm balloon caused more strictures (31%) than treatments using a 5-cm balloon (7%). Pretreatment of nodules or retreatment of skipped areas did not increase strictures. Patients receiving subsequent PDT had a higher incidence of strictures. Cancer was eliminated in all patients. High-grade dysplasia was eliminated in 41 of 43 patients (96%). Barrett's mucosa was totally eliminated in 25 of 60 patients (42%). CONCLUSIONS: Oral prednisone after PDT did not reduce the incidence of strictures. Subsequent PDT and longer balloon window were associated with higher incidence of strictures. PDT followed by thermal ablation of small islands eliminated dysplasia, early cancer, and Barrett's mucosa. PMID- 11007215 TI - Expandable metal stents in achalasia--is there a role? AB - OBJECTIVE: Achalasia is treated with pneumatic dilation or myotomy, and botulinum toxin injections are occasionally used. We review our community's experience with expandable metal stents in six patients who failed medical treatment or were poor surgical candidates. METHODS: Eight stents were placed in six patients between July 1995 and November 1997. Four patients had achalasia and two pseudoachalasia. Four patients underwent successive botulinum toxin injections. One patient only agreed to periodic Maloney dilatations or a stent. Pneumatic dilation was performed in one patient and considered high risk in the rest. All were poor surgical candidates. Three different stents were used: Gianturco Rosch Z stent, Wallstent I, and Wallstent II. RESULTS: One-month mortality and morbidity were 33% and 50%, respectively. Two patients were asymptomatic on a liquid diet for > or =6 months but required repeat endoscopy for recurrent dysphagia because of food bolus impaction and proximal stent migration in each. CONCLUSIONS: Expandable metal stents in achalasia or pseudoachalasia do not provide sustained symptom relief, and their use is associated with unacceptably high morbidity and mortality. We do not recommend the use of these devices in patients who have failed medical therapy or who are poor surgical candidates. PMID- 11007216 TI - Intraesophageal acid perfusion sensitizes the esophagus to mechanical distension: a Barostat study. AB - OBJECTIVE: The pathogenesis of noncardiac chest pain is unclear. Increased gastroesophageal reflux and decreased pain thresholds to intraesophageal balloon distension have been demonstrated in a proportion of such patients. We aimed to investigate whether acid exposure sensitizes esophageal mechanoreceptors in healthy volunteers. METHODS: After an overnight fast, an infinitely compliant balloon, 4.5 cm in length and mounted on a multilumen transnasal manometry catheter, was placed 8.5 cm above the lower esophageal sphincter in 12 healthy male volunteers aged 18-39 yr. After determination of the minimal distending pressure, the balloon was inflated up to 48 mm Hg by means of a computer controlled barostat (G & J Electronics, Canada). Graded stepwise distensions were interspersed with random decreases in pressure to two-thirds of the previous value. At each pressure level, the subjects were asked to report on sensation and the presence of pain. Baseline distension was repeated to determine reproducibility of the pressure/volume relationship and also the perception and pain thresholds. After the baseline distension sequence, the esophagus was perfused for 20 min (at 7 ml/min) with either normal saline (control) or 0.1 N hydrochloric acid at 37 degrees C on a random basis. RESULTS: Basal sensory thresholds varied widely (first perception 5-36 mm Hg, pain 8 > or = 43 mm Hg). Two subjects did not experience pain up to the maximum distending pressure (42 and 43 mm Hg, respectively, after correction for the minimal distending pressure). Esophageal body compliance was similar on repeat distension. Sensory thresholds were reproducible with different distensions (perception r = 0.99, pain r = 0.95). Saline resulted in no significant changes in perception or pain thresholds. Acid perfusion reduced first perception (median before and after acid, 15 mm Hg and 8 mm Hg, respectively, p = 0.05) and pain threshold (median before and after acid, 32.5 mm Hg and 26.5 mm Hg, respectively, p = 0.05). When compared to changes after saline perfusion, acid perfusion reduced the perception threshold (median change, -3.8 mm Hg vs 0 mm Hg, p = 0.04) and tended to reduce the pain threshold (median change, -3.75 mm Hg vs +0.75 mm Hg, p = 0.09). CONCLUSIONS: Intraesophageal balloon distension using a barostat is a reproducible method of measuring esophageal body compliance and sensory thresholds. Acute exposure to acid seems to sensitize the esophagus to perception from intraluminal balloon distension. PMID- 11007217 TI - Endoscopic and histological comparison of nonulcer dyspepsia with and without Helicobacter pylori infection evaluated by the modified Sydney system. AB - OBJECTIVE: Our aim was to identify endoscopic features associated with Helicobacter pylori (H. pylori) infection in patients with nonulcer dyspepsia. METHODS: A total of 50 infected patients with nonulcer dyspepsia who underwent endoscopy with antral and corporal biopsies and 50 patients matched for age and sex but with nonulcer dyspepsia without H. pylori were reviewed retrospectively by three endoscopists blinded to the H. pylori status and the patient's history. The endoscopic findings of gastritis, classified by a modification of the Sydney system as present or absent, were evaluated, and the histological severity was graded by the updated Sydney system. RESULTS: For endoscopic features, the odds ratio was 53.1 (95% confidence interval, 6.8-414.9) for edema, 18.8 (5.8-60.5) for erythema with reddish streaks excluded, 0.0275 (0.0002-0.477) for reddish streaks, 17.4 (0.97-313.7) for friability, 14.2 (5.1-40.0) for exudate, 17.2 (2.2 137.6) for flat erosions, 2.54 (0.81-7.94) for raised erosions, 40.1 (2.3-694.5) for rugal hypertrophy, 19.1 (2.4-151.6) for rugal atrophy, 96.2 (23.4-395.9) for a vascular pattern, 0.125 (0.010-1.06) for bleeding spots, and 21.0 (2.6-166.5) for nodularity. The histological severity of inflammation, neutrophil activity, and atrophy in the antrum and corpus and of metaplasia in the antrum was greater in the infected patients than in the noninfected patients. CONCLUSIONS: Endoscopic features associated with H. pylori were a vascular pattern, edema, rugal hypertrophy, nodularity, rugal atrophy, erythema with reddish streaks excluded, flat erosions, and exudate. These endoscopic features were associated with the histological findings of inflammation, neutrophil activity, atrophy, and metaplasia. PMID- 11007218 TI - Accuracy of an enzyme immunoassay for the detection of Helicobacter pylori in stool specimens in the diagnosis of infection and posttreatment check-up. AB - OBJECTIVE: The aim of this study was to assess the reliability of a newly developed enzyme immunoassay for Helicobacter pylori-specific antigen detection in stools (HpSA) compared to other standardized diagnostic techniques such as histology (H), rapid urease test (RUT) and 13C-urea breath test (UBT) to diagnose H. pylori infection and to evaluate its usefulness in determining H. pylori status after treatment. METHODS: One hundred eighty-eight patients referred to our department for upper gastrointestinal endoscopy were included. H. pylori infection was confirmed in all patients by HpSA test in stools, RUT, UBT, and H. Patients were defined as positive for H. pylori if RUT and UBT or H were positive. A total of 142 symptomatic patients received eradication treatment and were reassessed 6 wk after therapy; for 70 of these patients, stool samples were also collected at 24 h and 6 months after finishing eradication treatment. In the posttreatment follow-up, UBT was used as gold standard. RESULTS: The sensitivity of HpSA test for the diagnosis of H. pylori infection using a cut-off value of 0.130 was 89.5% and its specificity 77.8%. This specificity was lower than that obtained with UBT, H, and RUT. In the early follow-up the sensitivity of HpSA test was null. At 6 weeks and at 6 months post-treatment its sensitivity was 70.4% and 50% and its specificity was 81.6% and 79.3%, respectively. CONCLUSIONS: The HpSA stool test, using a cut-off value of 0.130, may be useful for the primary diagnosis of H. pylori infection, with sensitivity similar to that obtained with other standard tests, but with less specificity. HpSA test is not useful for early monitoring of treatment efficacy. At 6 wk and at 6 months posttreatment, HpSA test lacks accuracy as compared to UBT for evaluating the outcome of the eradication treatment. PMID- 11007219 TI - CagA-positive strains of Helicobacter pylori may protect against Barrett's esophagus. AB - OBJECTIVE: Helicobacter pylori (H. pylori) colonization is associated with chronic gastritis, peptic ulcer disease, and adenocarcinoma of the distal stomach. However, the role of H. pylori strain variation in complicated gastroesophageal reflux disease, especially Barrett's esophagus, is unknown. Therefore, the aim of this study was to evaluate the prevalence of colonization by cagA+ and cagA- H. pylori strains in the spectrum of gastroesophageal reflux disease, including Barrett's esophagus. METHODS: A total of 251 patients undergoing endoscopy were categorized into four groups: controls, patients with gastroesophageal reflux disease alone, and patients with short- and long-segment Barrett's esophagus. All patients underwent upper endoscopies with biopsies and serum collections. H. pylori and degree of mucosal inflammation in gastric biopsies were assessed and serological assessment made for H. pylori and cagA status. RESULTS: The overall prevalence of H. pylori colonization in the study population was 35% (95% confidence interval = 29.5-41.4%) which did not differ significantly among the groups. However, colonization by cagA+ H. pylori strains was significantly more prevalent among controls (11/25; 44%) and patients with gastroesophageal reflux disease (13/36; 36%) than in patients with short-segment (2/10; 20%) or long-segment Barrett's esophagus (0/18; 0%). Patients with Barrett's esophagus were less likely to be colonized by cagA+ H. pylori strains than reflux patients without Barrett's esophagus (odds ratio = 0.27, 95% confidence interval = 0.11-0.67, p = 0.004). CONCLUSIONS: Colonization by cagA+ H. pylori strains may be protective against the formation of short- and long segment Barrett's esophagus and its malignant complications. PMID- 11007220 TI - Treatment of uninvestigated dyspepsia with cisapride for patients with negative Helicobacter pylori serologies. AB - OBJECTIVE: The aim of this study was to compare symptoms for patients with uninvestigated dyspepsia and a negative Helicobacter pylori serology who were treated with cisapride or placebo. METHODS: Helicobacter pylori-seronegative patients with chronic dyspepsia were randomized to receive cisapride 10 mg t.i.d. or placebo t.i.d. for 30 days. Symptom scores were performed 1 month and 3 months after randomization. Outcomes measured were dyspepsia symptom scores and a treatment "success" variable defined as absence of symptoms or decrease in the most severe individual symptom by two grades. RESULTS: A total of 60 patients were randomized; 56 completed the 1-month follow-up and 40 completed the 3-month follow-up interview. The mean score for all patients at the time of entry was 11.0 and declined to 8.3 and 8.2 at 1 and 3 months, respectively, after randomization. At 1 month and 3 months after randomization, there was no significant difference in the number of patients meeting the "success" variable for patients receiving cisapride as compared to placebo. The mean decline in symptom severity scores was not significantly different for patients receiving placebo or cisapride at 1 month (mean, -2.8 vs -3.1; difference = 0.3, p = 0.74) or 3 months (-3.1 vs -2.6, difference = -0.5, p = 0.58) after randomization. CONCLUSIONS: No significant difference in the severity of dyspeptic symptoms was found for patients receiving cisapride as compared to placebo in the setting of uninvestigated dyspepsia and a negative Helicobacter pylori serology. PMID- 11007221 TI - Risk of upper gastrointestinal bleeding associated with use of low-dose aspirin. AB - OBJECTIVE: Aspirin products are known to cause irritation and injury to the gastric mucosa. We examined the risk of hospitalization for upper gastrointestinal bleeding with use of low-dose aspirin. METHODS: This was a cohort study based on record linkage between a population-based prescription database and a hospital discharge registry in North Jutland County, Denmark, from January 1, 1991, to December 31, 1995. Incidence rates of upper gastrointestinal bleeding in 27,694 users of low-dose aspirin were compared with the incidence rates in the general population in the county. RESULTS: A total of 207 exclusive users of low-dose aspirin experienced a first episode of upper gastrointestinal bleeding with admission to the hospital during the study period. The standardized incidence rate ratio was 2.6 (95% confidence interval, 2.2-2.9), 2.3 in women and 2.8 in men. The standardized incidence rate ratio for combined use of low-dose aspirin and other nonsteroidal anti-inflammatory drugs was 5.6 (95% confidence interval, 4.4-7.0). The risk was similar among users of noncoated low-dose aspirin (standardized incidence rate ratio, 2.6; 95% confidence interval, 1.8 3.5) and coated low-dose aspirin (standardized incidence rate ratio, 2.6; 95% confidence interval, 2.2-3.0). CONCLUSIONS: Use of low-dose aspirin was associated with an increased risk of upper gastrointestinal bleeding, with still higher risks when combined with other nonsteroidal anti-inflammatory drugs. Enteric coating did not seem to reduce the risk. The findings from this observational study raise the possibility that prophylactic use of low-dose aspirin may convey an increased risk of gastrointestinal bleeding, which may offset some of its benefits. PMID- 11007222 TI - Treatment of Helicobacter pylori in patients with duodenal ulcer hemorrhage--a long-term randomized, controlled study. AB - OBJECTIVE: Eradication of Helicobacter pylori (H. pylori) in patients with uncomplicated duodenal ulcers prevents long-term recurrence of ulcers. We aimed to study whether treatment of H. pylori prevents the long-term recurrence of duodenal ulcer hemorrhage. METHODS: Patients with duodenal ulcer bleeding and confirmed H. pylori infection were recruited. A total of 120 patients were randomly assigned to triple therapy (DeNoltab 120 mg, amoxycillin 500 mg, and metronidazole 300 mg four times daily) or DeNoltab 120 mg four times daily alone. No maintenance therapy was given during the follow-up period. The endpoints were the cumulative rates of symptomatic and bleeding duodenal ulcer recurrences. RESULTS: Of the patients receiving the triple regimen, 85.1% had H. pylori eradicated as compared to 2.0% of patients receiving DeNoltab (p < 0.05). More patients in the DeNoltab group than those in the Triple group had recurrence of ulcer bleeding, but this did not reach statistical significance (12/60 vs 6/60, p = 0.20). Logistic regression analysis on clinical, personal, and endoscopic characteristics identified persistent H. pylori infection as the only independent predictor of recurrence of duodenal ulcer bleeding. CONCLUSIONS: Treatment of H. pylori alone with the present bismuth-based triple therapy in patients with duodenal ulcer hemorrhage did not result in significant reduction in further bleeding episodes, although a trend was seen for the group that was given triple therapy. On the other hand, posttreatment H. pylori status was found to be an independent predictor of bleeding recurrence. PMID- 11007223 TI - The stimulation of antral motility by erythromycin is attenuated by hyperglycemia. AB - OBJECTIVE: Diabetic gastroparesis is usually treated with prokinetic drugs, of which the most potent, when given intravenously during euglycemia, is erythromycin. Recent studies have demonstrated that the gastrokinetic effects of erythromycin are attenuated by hyperglycemia. The aim of this study was to determine whether the effects of erythromycin on antropyloroduodenal motility, including the organization of antral pressure waves, are modified by hyperglycemia. METHODS: A total of eight healthy male volunteers (median age 24 yr) were studied on 2 days each in randomized order. A manometric assembly, incorporating six antral, two pyloric, and seven duodenal sideholes and a pyloric sleeve sensor, was positioned with the sleeve spanning the pylorus. The blood glucose concentration was stabilized at about 5 mmol/L (euglycemia) or 15 mmol/L (hyperglycemia). After 30 min (T = 0), an intraduodenal lipid infusion (1.5 kcal/min) was commenced and continued until the end of the study. At T = 20 minutes, erythromycin (200 mg) as the lactobionate was infused intravenously over 20 min, followed by 100 mg over the next 40 min. RESULTS: Intravenous erythromycin increased the amplitude of antral waves during intraduodenal lipid infusion at both blood glucose concentrations (p < 0.01 for euglycemia and p < 0.05 for hyperglycemia). After erythromycin (T = 20 to T = 80), the frequency (p < 0.05) and amplitude (p < 0.01) of antral waves were less during hyperglycemia than euglycemia. Both propagated (p < 0.0005) and nonpropagated (p < 0.01) antral waves were decreased by hyperglycemia, but the suppression of propagated waves was greater (p < 0.05). Erythromycin reduced the frequency (p = 0.09) but increased the amplitude (p < 0.05) of phasic pyloric pressures, and decreased basal pyloric pressure (p < 0.0005). The frequency (p = 0.06) and amplitude (p < 0.05) of phasic pyloric waves during erythromycin infusion were slightly less during hyperglycemia than euglycemia, whereas there was no effect of the blood glucose concentration on basal pyloric pressure. Erythromycin increased the amplitude (p < 0.001) but not the frequency of duodenal waves; the frequency and amplitude of duodenal waves did not differ between the two blood glucose concentrations. CONCLUSIONS: Hyperglycemia attenuates the stimulation of antral pressures and propagated antral sequences by erythromycin, but not the effects of erythromycin on pyloric or duodenal motility. PMID- 11007224 TI - Sedation for colonoscopy using a single bolus is safe, effective, and efficient: a prospective, randomized, double-blind trial. AB - OBJECTIVE: Practice guidelines call for the careful titration of sedatives and analgesics during endoscopy, with time taken between incremental doses to assess effect. This approach is time-consuming and has never been validated in a prospective trial. The aim of this study was to compare the safety and efficacy of titration, as outlined in practice guidelines, with a single, rapid bolus of sedatives before colonoscopy. METHODS: Consecutive colonoscopy outpatients were randomized to a single, rapid bolus of meperidine and midazolam or to a titration of doses every 3 min until predefined levels of somnolence were achieved. The colonoscopist was not present during sedation and remained blinded as to which technique was used. Supplemental O2 was given for SaO2 <90% on three or more occasions. Total physician time was calculated from the first injection of sedatives to the removal of the colonoscope. Patient assessments of pain and tolerance were obtained at the time of discharge using visual analog scales of 100 mm (0 = excellent and 100 = unbearable). RESULTS: A total of 101 patients were randomized (49 bolus, 52 titration). Demographic features were similar for both groups. Titration required more physician time than did bolus (32.2 min vs 20.1 min, p < 0.001) and was associated with an increased need for supplemental O2 (44% vs 14%, p = 0.002). Mean tolerance scores were similar (titration 16.3 vs bolus 15.3, p = 0.72). CONCLUSIONS: Rapid bolus sedation for colonoscopy saves significant endoscopist time, is associated with less O2 desaturation, and provides equivalent levels of patient comfort. A revision of the guidelines for sedation and analgesia during endoscopy should be considered. PMID- 11007225 TI - Impact of endoscopic ultrasound on the management and outcome of pancreatic carcinoma. AB - OBJECTIVE: Although endoscopic ultrasound and endoscopic ultrasound-guided fine needle aspiration have been shown to aid in the diagnosis and staging of pancreatic carcinoma, whether the general use of these new techniques impacts the overall management and survival of pancreatic cancer, is less clear. METHODS: Clinical data on all patients diagnosed with pancreatic carcinoma were assessed for two equal time periods during which computerized tomography-guided fine needle aspiration and biopsy was used (1/93-5/95) and then during which endoscopic ultrasound-guided fine needle aspiration was used (8/95-12/97) for the primary diagnosis of these patients. RESULTS: Comparative data for the computerized tomography era versus the endosonography era showed that 1) pancreatic carcinomas diagnosed: 52 versus 84 patients (stage I-III at diagnosis: 33% vs 54%; p < 0.05); 2) diagnosis by aspiration or biopsy of a pancreatic mass, nodes, liver metastasis, or by operation: 46%, 0%, 19%, 29% versus 52%, 8% (p = 0.02 nodes plus pancreatic masses), 20%, 7% (p = 0.002); 3) pancreatic resections: 13% versus 14%; 4) median survival without liver metastases: 102 versus 205 days (p < 0.02, log-rank test). Endoscopic ultrasound detected carcinomas that were either not seen or only possibly seen by computed tomography in 34%. CONCLUSIONS: More patients (62%) were diagnosed with pancreatic carcinoma when using endoscopic ultrasound and 75% fewer required operations for diagnosis. The 3-month improvement in median survival is probably multifactorial but most likely represents lead bias because of the greater sensitivity of endoscopic ultrasound for pancreatic carcinoma. Using endoscopic ultrasound with endoscopic ultrasound-guided fine needle aspiration in patients with pancreatic carcinoma significantly affects their management and outcomes. PMID- 11007226 TI - Endosonography-guided, fine-needle aspiration cytology extending the indication for organ-preserving pancreatic surgery. AB - OBJECTIVE: Organ preserving pancreatic resections are considered whenever malignant disease is ruled out. In tumors of low malignant potential such as cystadenomas and neuroendocrine tumors, the diagnosis is rarely established preoperatively. We studied the feasibility of cytodiagnosis using endosonography guided fine-needle aspiration in determining the operative approach. METHODS: A total of 78 patients (16 female, 62 male; mean age 61.4 yr, range 31-82 yr) with focal pancreatic lesions underwent EUS-FNA. Final diagnosis was confirmed by histology, cytology, or clinical follow up (>9 months). Patients with tumors of low malignant potential were managed by customized pancreatic resections. RESULTS: Final diagnosis was malignant tumors in 36 patients, tumors of low malignant potential in nine (six, neuroendocrine, two, borderline mucinous cystadenomas, one, borderline adenocarcinoma), and benign in 31 (two inadequate smears). No complications occurred. With six false-negative and no false-positive results, the accuracy, sensitivity, specificity, and positive and negative predictive values were 92%, 84%, 100%, 100%, and 86%, respectively. Five patients with low malignant tumors underwent duodenum-preserving pancreatic head resection, three mid segment resection, and one pylorus-preserving pancreatoduodenectomy. CONCLUSIONS: EUS-FNA is useful in the preoperative cytodiagnosis of pancreatic tumors of low malignant potential. It extends the indication for organ-preserving pancreatic resections and avoids the unnecessary sacrifice of adjacent organs. PMID- 11007227 TI - Modern imaging methods versus clinical assessment in the evaluation of hospital in-patients with suspected pancreatic disease. AB - OBJECTIVE: Various modern imaging procedures such as endoscopic retrograde cholangiopancreatography (ERCP), computed tomography (CT), and endoscopic ultrasonography (EUS) have been shown to be highly accurate in the diagnosis of specific disorders of the pancreas. However, prior information often causes bias in the interpretation of these results. Little information is available concerning the value of these examinations in the primary and differential diagnosis of suspected pancreatic disease-particularly in comparison with clinical evaluation, including laboratory tests and transabdominal ultrasound (TUS). METHODS: Clinical and imaging information (EUS, ERCP, and CT) was collected for 184 inpatients who were referred over a 5-yr period for evaluation of suspected pancreatic disease. On the basis of patient history, laboratory tests, and the results of routine TUS, one gastroenterologist, who was unaware of any of the other procedures or the final diagnosis, made a presumptive clinical diagnosis. CT and ERCP images and EUS videotapes were then analyzed by three different and independent examiners, who had the same clinical information except for the TUS results, but were completely blinded to the results of the other examinations and the patients' diagnoses. The final diagnoses were obtained by surgery, histology, and cytology, plus a follow-up of at least 1 yr (mean 35 months) in all noncancer cases. RESULTS: The final diagnoses were: normal pancreas (n = 36), chronic pancreatitis without a focal inflammatory mass (n = 53) or with a focal inflammatory mass (n = 18), and pancreatic malignancy (n = 77). Clinical evaluation, including ultrasonography, achieved a sensitivity for pancreatic disease of 94% but a specificity of only 35%. The figures for the sensitivity and specificity of the three imaging procedures were 93% and 94%, respectively, for EUS; 89% and 92% for ERCP; and 91% and 78% for CT (p < 0.05 for the specificity of clinical assessment vs all three imaging tests, p > 0.05 for comparison of the three imaging procedures). In the differential diagnosis between cancer and chronic pancreatitis as well as between malignant and inflammatory tumors, there was no difference among clinical assessment and the three imaging tests. CONCLUSIONS: In a group of patients with a high suspicion of pancreatic disease, little additional sensitivity in the diagnosis of pancreatic disease is provided by sophisticated imaging procedures such as EUS, ERCP, and CT, in comparison with clinical assessment including laboratory values and TUS. However, the specificity can be substantially improved. To confirm the diagnosis, one of the three examinations is needed, depending on the suspected disease and local expertise. The imaging procedures should be performed in a stepwise fashion for specific purposes, such as exclusion of pancreatic disease and the planning of treatment in chronic pancreatitis and pancreatic cancer. PMID- 11007228 TI - Endoscopic ultrasound is highly accurate and directs management in patients with neuroendocrine tumors of the pancreas. AB - Preoperative localization of pancreatic neuroendocrine tumors with traditional imaging fails in 40-60% of patients. Endoscopic ultrasound (EUS) is highly sensitive in the detection of these tumors. Previous reports included relatively few patients or required the collaboration of multiple centers. We report the results of EUS evaluation of 82 patients with pancreatic neuroendocrine tumors. METHODS: We prospectively used EUS early in the diagnostic evaluation of patients with biochemical or clinical evidence of neuroendocrine tumors. Patients had surgical confirmation of tumor localization or clinical follow-up of >1 yr. RESULTS: Eighty-two patients underwent 91 examinations (cases). Thirty patients had multiple endocrine neoplasia syndrome type 1. One hundred pancreatic tumors were visualized by EUS in 54 different patients. The remaining 28 patients had no pancreatic tumor or an extrapancreatic tumor. Surgical/pathological confirmation was obtained in 75 patients. The mean tumor diameter was 1.51 cm and 71% of the tumors were < or =2.0 cm in diameter. Of the 54 explorations with surgical confirmation of a pancreatic tumor, EUS correctly localized the tumor in 50 patients (93%). Twenty-nine insulinomas, 18 gastrinomas, as well as one glucagonoma, one carcinoid tumor, and one somatostatinoma were localized. The most common site for tumor localization was the pancreatic head (46 patients). Most tumors were hypoechoic, homogenous, and had distinct margins. EUS of the pancreas was correctly negative in 20 of 21 patients (specificity, 95%). EUS was more accurate than angiography with or without stimulation testing (secretin for gastrinoma, calcium for insulinoma), transcutaneous ultrasound, and CT in those patients undergoing further imaging procedures. EUS was not reliable in localizing extrapancreatic tumors. CONCLUSIONS: In this series, the largest single center experience reported to date, EUS had an overall sensitivity and accuracy of 93% for pancreatic neuroendocrine tumors. Our results support the use of EUS as a primary diagnostic modality in the evaluation and management of patients with neuroendocrine tumors of the pancreas. PMID- 11007229 TI - Mediastinal lymphadenopathy in patients with or without previous malignancy: EUS FNA-based differential cytodiagnosis in 153 patients. AB - OBJECTIVE: Mediastinal lymphadenopathy (ML) is a cause for concern, especially in patients with previous malignancy. The investigation of choice is thoracic CT with a variable sensitivity and specificity requiring tissue diagnosis. We used endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for cytodiagnosis of ML in patients with and without previous malignancy. The cause, distribution of lesions, and incidence of second cancers were investigated. METHODS: Linear echoendoscopes and 22-gauge needles for cytology were used for EUS-FNA. A cytological diagnosis of malignancy was accepted, and histology or consistent follow-up of at least 9 months confirmed benign results. RESULTS: One hundred fifty-three patients underwent EUS-FNA between November 1997 and November 1999 (mean age, 60 yr; range, 13-82 yr; 105 men). Cytology was adequate in 150 patients. Final diagnosis was malignancy in 84 and benign in 66 patients (sensitivity, specificity, and diagnostic accuracy: 92%, 100%, 95%, respectively). In 101 patients without previous cancer cytology identified 48 malignant (lung, 41; extrathoracic, 7) and 51 benign lesions (inflammation, 35; various, 9; sarcoidosis, 7) (sensitivity, specificity, accuracy: 88%, 100%, 94%). Fifty-two patients had prior malignancy, mostly in extrathoracic sites. Cytology revealed recurrences in 21 patients, second cancer in 9 and benign lesions in 21 patients (inflammatory, 11; sarcoidosis, 8; tuberculosis, 1; abscess, 1) (sensitivity, specificity, accuracy: 97%, 100%, 98%). CONCLUSIONS: In patients without previous cancer malignant ML originates from the lung >80%. In those with previous malignancy recurrence of extrathoracic sites is the major cause. Benign lesions and treatable second cancers occur in a significant frequency, emphasizing the need for tissue diagnosis. EUS-FNA is a safe and minimally invasive alternative for cytodiagnosis in the mediastinum. PMID- 11007230 TI - Enterocolitis in children with developmental disorders. AB - OBJECTIVE: Intestinal pathology, i.e., ileocolonic lymphoid nodular hyperplasia (LNH) and mucosal inflammation, has been described in children with developmental disorders. This study describes some of the endoscopic and pathological characteristics in a group of children with developmental disorders (affected children) that are associated with behavioral regression and bowel symptoms, and compares them with pediatric controls. METHODS: Ileocolonoscopy and biopsy were performed on 60 affected children (median age 6 yr, range 3-16; 53 male). Developmental diagnoses were autism (50 patients), Asperger's syndrome (five), disintegrative disorder (two), attention deficit hyperactivity disorder (ADHD) (one), schizophrenia (one), and dyslexia (one). Severity of ileal LNH was graded (0-3) in both affected children and 37 developmentally normal controls (median age 11 yr, range 2-13 yr) who were investigated for possible inflammatory bowel disease (IBD). Tissue sections were reviewed by three pathologists and scored on a standard proforma. Data were compared with ileocolonic biopsies from 22 histologically normal children (controls) and 20 children with ulcerative colitis (UC), scored in an identical manner. Gut pathogens were sought routinely. RESULTS: Ileal LNH was present in 54 of 58 (93%) affected children and in five of 35 (14.3%) controls (p < 0.001). Colonic LNH was present in 18 of 60 (30%) affected children and in two of 37 (5.4%) controls (p < 0.01). Histologically, reactive follicular hyperplasia was present in 46 of 52 (88.5%) ileal biopsies from affected children and in four of 14 (29%) with UC, but not in non-IBD controls (p < 0.01). Active ileitis was present in four of 51 (8%) affected children but not in controls. Chronic colitis was identified in 53 of 60 (88%) affected children compared with one of 22 (4.5%) controls and in 20 of 20 (100%) with UC. Scores of frequency and severity of inflammation were significantly greater in both affected children and those with UC, compared with controls (p < 0.001). CONCLUSIONS: A new variant of inflammatory bowel disease is present in this group of children with developmental disorders. PMID- 11007231 TI - Do male sex hormones protect from irritable bowel syndrome? AB - OBJECTIVE: Irritable bowel syndrome (IBS) is more common in women and it is frequently assumed that being female may predispose to the development of this disorder. Alternatively, being male could offer some degree of protection and if so, this might be mediated by testosterone. The aim of this study was to assess whether male patients with IBS have lower levels of testosterone and related gonadotrophins than their unaffected counterparts and if this relates to rectal sensitivity. METHODS: Fifty secondary care, male outpatients with IBS (aged 19-71 yr) were compared with 25 controls (aged 22-67 yr). Each subject had serum testosterone, free testosterone, sex hormone-binding globulin, follicle stimulating hormone, and luteinizing hormone (LH) measured, together with rectal sensitivity to balloon distension. Anxiety and depression were also assessed. RESULTS: The only difference in the hormone levels between patients and controls that reached statistical significance was the lower value for LH in the IBS patients (p = 0.014). Although patients were more anxious and depressed than the controls (p < 0.001), this could not solely account for the reduced level of LH, as adjusting for these (analysis of variance) still tended to show that LH values were lower in men with rather than without IBS [F(1,70) = 2.74; p = 0.10]. Men with IBS were more sensitive to balloon distension of the rectum, with the distension volumes required for "urgency" (p < 0.001) and "discomfort" (p = 0.001) significantly lower than controls. Paradoxically, the patient's sensory thresholds negatively correlated with levels of testosterone (p < 0.05) and free testosterone (p < 0.002), and positively with levels of sex hormone-binding globulin (p < 0.05). Finally, there was a tendency for IBS symptomatology to be inversely related to testosterone levels (p = 0.15). CONCLUSIONS: These results support the need for further exploration of the role of male sex hormones in the pathophysiology of IBS. PMID- 11007232 TI - High frequencies of telomeric associations, chromosome aberrations, and sister chromatid exchanges in ulcerative colitis. AB - OBJECTIVE: Chromosome instability provides a predisposing background to malignancy, contributing to the crucial genetic changes in multistep carcinogenesis. The aim of this work was to analyze chromosome instability in patients with ulcerative colitis (UC) to achieve a better understanding of the increased risk for colorectal cancer. METHODS: Peripheral blood lymphocyte cultures from 20 untreated UC patients and 24 controls were used to study chromosome instability by assessing telomeric associations (TAS), chromosome aberrations (CA), and sister chromatid exchanges (SCE). RESULTS: Mean frequencies of TAS and CA were significantly increased in UC patients compared to controls (p < 0.001). Chromosomes 10, 11, 21, 16, and 19 were the most frequently involved in TAS. A total of 104 CA clustered in 66 breakpoints could be exactly localized. Seven nonrandom bands significantly affected in UC patients were found (p < 0.004), showing a significant correlation with the location of cancer breakpoints (p < 0.003), particularly with colorectal carcinoma rearrangements. SCE analysis showed higher levels in patients compared to controls (p < 0.006), but no differences were observed in cell cycle kinetics. CONCLUSIONS: Our results demonstrate the presence of an unstable genome in UC patients that could be related to the cancer development observed in this disease. PMID- 11007233 TI - Risk of lymphoma in inflammatory bowel disease. AB - OBJECTIVE: Inflammatory bowel disease with a subsequent diagnosis of non Hodgkin's lymphoma has been reported. There is concern that the risk of developing lymphoma will rise with increasing use of immune modifier therapy. We determined the risk of non-Hodgkin's lymphoma in an U.S. population-based inception cohort, and evaluated the association between inflammatory bowel disease and lymphoma in our referral practice. METHODS: The records of all incidence cases of inflammatory bowel disease in Olmsted County, Minnesota, between 1950 and 1993 were reviewed for the diagnosis of lymphoma. Standardized incidence ratios (observed/expected) were used to estimate relative risk. In addition, the clinical features and outcomes of all patients with inflammatory bowel disease and lymphoma seen at Mayo Clinic between 1976 and 1997 were reviewed. RESULTS: Among 454 county residents diagnosed with inflammatory bowel disease, a single non-Hodgkin's lymphoma occurred in a patient with Crohn's disease. No cases were seen with ulcerative colitis. The estimated relative risk of lymphoma was 2.4 in Crohn's disease (95% confidence interval, 0.1-13), 0 in ulcerative colitis (0-6), and 1.0 in inflammatory bowel disease overall (0.03-6). Between 1976 and June 1997, 61 patients with inflammatory bowel disease and lymphoma (approximately 0.41%) were seen in the referral practice. In four patients with Crohn's disease (13%), potential neoplastic risk factors were identified-therapeutic radiation in 1, and use of purine analogs in 3 (median length of use, 11 months). CONCLUSIONS: Our population-based cohort study demonstrated that the absolute risk of non-Hodgkin's lymphoma remains quite small (0.01% per person-year). This risk may not exceed that in the general population. In our referral practice, immune modifier therapy could be potentially implicated in only 5% of cases of lymphoma occurring in the setting of inflammatory bowel disease. PMID- 11007234 TI - Allelic variants of the thiopurine S-methyltransferase deficiency in patients with ulcerative colitis and in healthy controls. AB - OBJECTIVE: Thiopurine S-methyltransferase (TPMT) is a cytosolic enzyme that catalyzes the inactivation of mercaptopurine, azathioprine, and thioguanine. The genetic polymorphisms in the TPMT gene that regulate TPMT activity are inherited as an autosomal recessive trait and patients with genetically determined low levels of TPMT activity develop severe myelosuppression when treated with standard doses of the above-mentioned drugs. We have analyzed the frequencies of the allelic variants of the TPMT gene in a white European population of healthy blood donors from Spain and The Netherlands, and in a group of patients suffering from ulcerative colitis (UC) with a similar genetic background. METHODS: Two hundred and thirteen unrelated healthy individuals (HC) and 146 UC patients were typed for the polymorphic sites at positions 460 (G-->A) and 719 (A-->G) of the TPMT gene using specific polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) methods. RESULTS: There were no significant differences between the allele frequencies observed in the group of UC patients and those of the control group (10% of cases were heterozygous carriers of a TPMT mutant allele). The most frequent mutant allele in both UC and HC groups was TPMT3A (A460-->G719) (60% of carriers). TPMT3B (A460-->A719) and TPMT3C (G460-->G719) alleles were more often found in our study than in previously reported studies, reflecting the different genetic backgrounds of the European populations analyzed. CONCLUSIONS: Genotyping methods provide a simple and reliable screening to identify patients with a high risk of developing severe bone marrow toxicity if treated with thiopurine drugs. In UC patients, TPMT genotype should be determined before the initiation of azathioprine therapy. PMID- 11007236 TI - Body composition changes induced by chronic ethanol abuse: evaluation by dual energy X-ray absorptiometry. AB - OBJECTIVE: Nutritional disorders in alcoholics remain one of the most relevant medical problems in Western societies. As ethanol can supply >50% of the dietary energy in alcoholics, body composition alterations may easily occur. The aim of the present study was to evaluate the influence of chronic alcohol consumption on body composition in alcoholics compared to healthy social drinkers. METHODS: A total of 34 alcoholics defined according to DSM III R criteria, aged 41.6 +/- 9.3 yr and with a body mass index (BMI) 23.8 +/- 3.2 kg/m2, were consecutively enrolled in the study. In addition, 43 healthy male social drinkers were used as controls. Body composition was assessed using dual energy x-ray absorptiometry (DXA), and dietary habits were determined by a 3-day food diary. RESULTS: Mean daily alcohol intake was 194 +/- 62.4 g/day in alcoholics and 35.7 +/- 5.2 in healthy subjects (p < 0.0001). Body weight did not differ between alcoholics and controls (70.1 +/- 9.9 vs 71.8 +/- 6.4 kg). Alcoholics had a lower percent body fat (PBF) than control subjects (18.7 +/- 3.7 vs 23.9 +/- 3.9%; p < 0.01), as well as a lower fat mass content (13.4 +/- 3.8 vs 17.0 +/- 3.7 kg; p < 0.01). BMI was highly correlated with PBF in the patient population studied (R = 0.79; p < 0.0001). Significantly higher waist-to-hip ratios were found in alcoholics than in healthy subjects (p < 0.01). No correlation was found between dose of ethanol or duration of alcohol abuse and any of the variables examined. CONCLUSIONS: Alcoholics showed a reduced fat mass and a good preservation of lean body mass with respect to control subjects, and duration of alcohol use and alcohol dose did not seem to influence body composition. These data suggest that, unlike control subjects, alcoholics cannot store the calories provided by ethanol as fat deposits. PMID- 11007235 TI - Tissue transglutaminase antibodies in celiac disease: assessment of a commercial kit. AB - OBJECTIVE: Tissue transglutaminase was identified as the autoantigen eliciting endomysial antibody. A homemade enzyme-linked immunosorbent assay (ELISA)-based test was recently developed to determine quantitative titers of IgA antitissue transglutaminase antibody. Our objective in this study was to assess the suitability of a newly developed commercial kit for quantitative determination of antibody in patients with untreated celiac disease. MATERIALS: We tested serum samples from 79 untreated celiac patients, 42 healthy blood donors, and 18 patients with nonceliac intestinal disorders evaluated in two different centers. Samples were tested for antitissue transglutaminase, and antiendomysial and antigliadin antibodies in the center where diagnosis was performed. To assess interlaboratory variability of methods, 24 samples randomly selected were blindly tested in both centers. Antitissue transglutaminase antibodies were determined using a commercial kit (INOVA Diagnostics, Inc., San Diego, CA). RESULTS: Untreated celiac patients had significantly higher titers of antitissue transglutaminase than healthy and disease controls (p < 0.00001). According to the cut-off provided by the manufacturers (20 AU/mL), overall sensitivity was 92% (85% for one center and 100% for the other) and specificity was 98% (100% and 95%, respectively). Antiendomysial antibody was 86% sensitive and 100% specific. Discordance between antitissue transglutaminase and antiendomysial antibodies was detected in 13% of patients. Although two antitissue transglutaminase-negative cases had a positive antiendomysial antibody, the inverse situation was found in eight cases. A blind determination of antitissue transglutaminase on the same samples evidenced a good agreement (kappa statistic: 0.66) between both centers when assessment was qualitative (based on the decision of positive or negative). Although correlation of titers for both determinations was highly significant (r: 0.902, p < 0.00001), a very wide interlaboratory variability (median: 50%) was detected when absolute values were considered. CONCLUSIONS: The quantitative determination of antitissue transglutaminase using a commercial kit was highly sensitive and specific for detection of celiac disease. We observed an incomplete overlapping with antiendomysial antibody. The very high variability of values between laboratories still remains to be solved so as to propose the commercial ELISA assay for the screening of celiac disease. PMID- 11007237 TI - Need for validation of clinical decision aids: use of the AST/ALT ratio in predicting cirrhosis in chronic hepatitis C. AB - OBJECTIVE: A value of > or = 1 for the ratio of aspartate amino-transferase to alanine aminotransferase (the AST/ALT ratio or AAR) has been shown to have a positive predictive value of 100% for the diagnosis of cirrhosis in patients with chronic hepatitis C. If validated on separate cohorts, an AAR > or = 1 might obviate the need for liver biopsy in some patients with hepatitis C. METHODS: We attempted to validate the AAR by abstracting demographic and clinical data from a database of consecutive patients with hepatitis C who had a liver biopsy between 1993 and 1998. We used definitions, methods of data collection, and analyses comparable to those of the published study. A hepatopathologist blindly reviewed 49 liver biopsies for histological grade and stage. RESULTS: The current cohort of 177 patients and the previous cohort of 139 patients were comparable in mean age (42.3 vs 43.8 yr), percentage of men (63 vs 67), percentage with an AAR > or =1 (20 vs 17), and Child-Pugh distribution, but differed in substantial use of ethanol (11% vs 3.6%; p = 0.01) and in the prevalence of cirrhosis (23% vs 34%, p = 0.06). Respective sensitivities of the AAR were 56% and 53%. An AAR > or =1 had a positive predictive value of 64% (95% confidence interval 48-78%) for the current cohort. Thirteen of 36 patients (36%) with an AAR > or =1 were incorrectly identified as having cirrhosis. Of these 13 patients, 6 had a normal AST and ALT, 5 had a minimally elevated AST or ALT, and 1 had advanced fibrosis without cirrhosis. CONCLUSIONS: These results suggest that an AAR > or =1 may not be as useful for predicting cirrhosis in chronic hepatitis C as previously thought, and emphasizes the need for validation of clinical decision aids on independent patient cohorts. PMID- 11007238 TI - Oral budesonide in the treatment of primary sclerosing cholangitis. AB - OBJECTIVE: This study was designed to evaluate the safety and estimate the efficacy of oral budesonide in patients with primary sclerosing cholangitis (PSC). METHODS: Twenty-one patients with PSC were treated with 9 mg daily of oral budesonide for 1 yr. RESULTS: Significant, but marginally important, improvement in serum alkaline phosphatase (1,235 +/- 190 vs 951 +/-206 U/L, p = 0.003) and AST levels (119 +/- 14 vs 103 +/- 19 U/L, p = 0.02) was noted at the end of the treatment period. Serum bilirubin levels increased significantly in the 18 patients who completed 1 yr of treatment (1.1 +/- 0.1 vs 1.4 +/- 0.3, p = 0.01) and no significant changes in liver tests were noted 3 months after budesonide was discontinued. The Mayo risk score did not change significantly, and although a significant improvement in the degree of portal inflammation was noted at the end of the treatment period, the degree of fibrosis and stage of disease were not significantly affected. There was a marked loss of bone mass of the femoral neck (0.851 +/- 0.02 vs 0.826 +/- 0.02 g/cm2, p = 0.002) and lumbar spine (1.042 +/- 0.02 vs 1.029 +/- 0.02 g/cm2, p = 0.09) at 1 yr of treatment with budesonide. Two patients required evaluation for liver transplantation during treatment, and two patients developed cosmetic side effects. CONCLUSIONS: Oral budesonide appears to be of minimal, if any, benefit and it is associated with a significant worsening of osteoporosis in patients with PSC. PMID- 11007239 TI - A pilot study of pentoxifylline for the treatment of primary sclerosing cholangitis. AB - OBJECTIVE: There is no effective therapy for patients with primary sclerosing cholangitis (PSC). Rats with experimental small bowel bacterial overgrowth develop hepatobiliary injury similar to PSC. The hepatobiliary injury results from peptidoglycan-polysaccharide-mediated activation of Kupffer cells, release of cytokines such as tumor necrosis factor (TNF-alpha), and is prevented by pentoxifylline. Our aims were to assess the safety and effects of pentoxifylline on symptoms and biochemical liver tests in patients with PSC. METHODS: A total of 20 patients with clinical, cholangiographic, and histological features of PSC of varying severity were treated with pentoxifylline sustained release (SR) tablets (400 mg q.i.d.) for < or = 1 yr. Serum alkaline phosphatase, aspartate aminotransferase, and bilirubin were monitored every 3 months for 1 year; serum TNF-alpha and TNF receptor subtypes I and II were assessed at baseline and 1 year. RESULTS: Of 20 patients, 16 tolerated pentoxifylline and completed the study. Two patients were withdrawn because of severe nausea, and two patients were noncompliant with medication and withdrew. Pentoxifylline did not significantly alter symptoms of fatigue or pruritus, serum liver tests, serum TNF alpha or TNF receptor levels. CONCLUSIONS: In the current regimen, pentoxifylline alone does not significantly improve symptoms or liver tests in patients with PSC. PMID- 11007240 TI - Bone density and metabolism in patients with viral hepatitis and cholestatic liver diseases before and after liver transplantation. AB - OBJECTIVE: Osteoporosis is frequently found in patients with cholestatic liver disease (primary biliary cirrhosis/primary sclerosing cholangitis) and chronic viral hepatitis. There is limited information about the long-term effect of liver transplantation (OLT) on bone metabolism. The aim of this study was to investigate the effect of liver transplantation on bone metabolism in patients with cholestatic and viral liver diseases. METHODS: We randomly recruited 193 patients with chronic viral hepatitis or cholestatic liver diseases. Bone density (Z-score) and markers of bone metabolism (intact parathyroid hormone [iPTH], PTH 70-84, osteocalcin, procollagen, telopeptide, and vitamin D) were determined before and at time points (< and > 24 months) post-OLT. RESULTS: Before OLT, bone density (Z-score) was decreased in patients with cholestatic (-1) and viral ( 0.4) liver diseases. In both groups bone density continued to decrease in the periods up to and more than 24 months after OLT. In the cholestatic group, bone density decreased significantly compared to pre-OLT (p < 0.05) and to the viral hepatitis group (p < 0,001). Markers of bone metabolism showed that after OLT, bone metabolism was enhanced and shifted versus bone resorption. Immunosuppressive drug therapy (glucocorticoids, cyclosporin, FK 506) directly correlated with increased bone metabolism post-OLT. CONCLUSIONS: Bone loss is a long-term problem after OLT, particularly in patients with cholestatic liver diseases. Drug therapy is a main factor of bone loss. Pre- and post-OLT therapy to reduce bone loss is recommended. PMID- 11007241 TI - Changing pattern of esophageal cancer incidence in New Mexico. AB - OBJECTIVE: Multiple reports indicate that esophageal adenocarcinoma incidence has increased during the past 20 yr, especially in non-Hispanic white men. We retrospectively reviewed adenocarcinoma and squamous cell carcinoma cases in our heterogeneous state population to determine the effect of ethnicity on histology. METHODS: We searched the New Mexico Tumor Registry for all cases of esophageal cancer from 1973 to 1997. Inclusion criteria included histological diagnosis of adenocarcinoma or squamous cell carcinoma, self-reported ethnicity, and gender. Age-adjusted incidence rates for both adenocarcinoma or squamous cell carcinoma were compared among ethnic groups in 5-yr intervals. RESULTS: Six hundred fifteen patients met inclusion criteria. Esophageal adenocarcinoma age-adjusted incidence rates/100,000 increased significantly during the 25-yr period: 1973-1977, 0.25 cases; 1978-1982, 0.33 cases; 1983-1987, 0.45 cases; 1988-1992, 0.85 cases; and 1993-1997, 1.19 cases; p < 0.001. In comparison, squamous cell carcinoma age adjusted incidence rates did not increase significantly during the study period. In non-Hispanic whites, the histological age-adjusted incidence rate changed during the 1993-1997 period compared to other periods: 1993-1997, squamous cell carcinoma 1.01 and adenocarcinoma 1.42, p < 0.001. In Hispanics, the age-adjusted incidence rate of adenocarcinoma increased significantly in the fifth period compared to other periods, p < 0.001. In all minority groups, squamous cell carcinoma remained the predominant type. CONCLUSIONS: Esophageal adenocarcinoma age-adjusted incidence increased in New Mexico from 1973 to 1997. This increase was found in non-Hispanic whites and Hispanics and became predominant in non Hispanic whites. Squamous cell carcinoma remains the primary type in minorities. This study suggests that ethnicity may influence esophageal cancer histology or ethnic background may place an individual at increased risk for certain types of esophageal cancer. PMID- 11007242 TI - Nonvariceal upper gastrointestinal bleeding: differences in outcome for patients admitted to internal medicine and gastroenterological services. AB - OBJECTIVE: It has been suggested that admission to a gastroenterology service (GAS) is associated with a better prognosis and lower cost for treatment of gastrointestinal (GI) diseases, such as upper GI bleeding (UGB). However, a large potential bias by higher comorbidity on internal medicine services (MED) could not be excluded from these studies. We therefore compared patients with upper GI bleeding admitted to a gastroenterology or internal medicine department, with special emphasis on prognostic factors, such as comorbidity, and outcome. METHODS: Between 1991 and 1995, 322 patients were admitted to our hospital for UGB. Forty-five patients had variceal and 277 patients had nonvariceal upper GI bleeding (NUGB). Of 232 patients with primary NUGB, 125 were admitted to GAS and 93 to MED. The charts of these patients were revised, comorbidity was carefully recorded, and the Rockall risk score was calculated. All deaths were individually classified as unavoidable, mostly due to severe underlying illness, or potentially avoidable. RESULTS: No differences in delay for endoscopy or treatment were observed between GAS and MED. The rebleeding, surgery, and mortality rates in GAS and MED patients were 11.6% versus 11.5% (NS), 7.8% versus 7.3% (NS), and 2.4% versus 10.8% (p = 0.02), respectively. Rockall scores differed between GAS and MED patients (3.1 +/- 1.8 vs 3.7 +/- 1.7, p = 0.02). The mortality rate stratified by Rockall score was lower for the GAS patients. However, individual analysis revealed that only three of 13 deaths were potentially avoidable: two of 10 at the MED and one of three at the GAS. CONCLUSION: The lower mortality among nonvariceal upper GI bleeding patients admitted to a gastroenterological service compared to an internal medicine service was mainly due to lesser comorbidity. This effect was not detected by stratification according to Rockall, but shown with analysis of individual patient charts only. The latter underscores the potential pitfalls when comparing outcome or cost of treatment between different medical services. PMID- 11007243 TI - Pill silverer. PMID- 11007244 TI - Microsporidial AIDS cholangiopathy due to Encephalitozoon intestinalis: case report and review. AB - Microsporidia are increasingly recognized as opportunistic infections in immunodeficient patients, predominantly patients with AIDS. The two microsporidia most commonly associated with disease in AIDS patients are Enterocytozoon bieneusi and Encephalitozoon intestinalis (previously known as Septata intestinalis). The most common clinical presentation of microsporidiosis in AIDS patients is diarrhea, most commonly caused by the Enterocytozoon bieneusi species. Encephalitozoon intestinalis is a recently described species that has been reported to cause disseminated human infection including cholangitis. We report a case of AIDS cholangiopathy that presented with abdominal pain and cholestatic liver tests. Ultrasound examination and ERCP revealed a picture of sclerosing cholangitis. Bile samples obtained at ERCP were negative for microsporidia; stool studies for microsporidia and cryptosporidia were also negative. No organisms were identified on routine light microscopy of the biopsy specimens from the duodenum, ampulla, and bile duct. E. intestinalis spores were demonstrated in the bile duct biopsies, by methylene blue and azure 11 staining and confirmed by electron microscopy. Albendazole therapy was successful in eradicating E. intestinalis with clinical improvement and improvement in CD4 count. However, the cholangiographic picture did not improve and repeat cholangiography revealed progressive bile duct injury. Albendazole therapy was delayed and may have been too late to prevent bile duct damage; the drug had to be approved by the US Food and Drug Administration for compassionate use. This is an unusual case of sclerosing cholangitis caused by an unusual organism and requiring biliary sphincterotomy and stent placement for progressive stricturing despite eradication of the infection. PMID- 11007245 TI - Solitary pelvic nodule: diagnosis of metastatic prostate cancer by endoscopic ultrasound-guided, fine-needle aspiration. AB - Prostate cancer manifesting as an isolated perirectal mass is a rare occurrence. The following is a report of a single pelvic nodule that was determined to be metastatic prostate cancer by endoscopic ultrasound-guided, fine-needle aspiration. PMID- 11007246 TI - Liver biopsies in adult hemophiliacs with hepatitis C: a United States center's experience. AB - We report a case series of 13 adult hemophiliacs with serological evidence of hepatitis C who underwent percutaneous liver biopsies without major complications. We also briefly review the recent literature on safety of liver biopsies in this population, and conclude that these patients may be safely biopsied using appropriate precautions regardless of the severity of hemophilia. PMID- 11007248 TI - Algorithm for the treatment of Crohn's disease: incorporation of emerging therapies. PMID- 11007247 TI - A case of primary hepatic lymphoma with hepatitis C liver cirrhosis. AB - Primary hepatic lymphoma is rare. The usual type is a large-cell, high-grade malignant B-cell lymphoma, although T-cell types have been described. Several cases of primary hepatic lymphoma of B-cell origin developing in patients with chronic hepatitis C virus infection have been reported. Recently, new findings have raised the question of the induction of lymphoma by hepatitis C virus. However, the causal relationship between hepatitis C viral infection and primary hepatic lymphoma remains obscure. This article reports a case of histologically proven primary hepatic lymphoma of T-cell origin, which was confined to the liver, in the setting of hepatitis C liver cirrhosis. This association has not previously been reported. PMID- 11007249 TI - Further evolution of hepatitis C treatment: individualized therapy. PMID- 11007250 TI - Re: Kilgore et al., The gastric cardia: fact or fiction? PMID- 11007251 TI - 6MP for fathers with IBD at the time of conception: the value of a large clinical experience over unrelated controls. PMID- 11007252 TI - Successful nonsurgical treatment of primary mucosa-associated lymphoid tissue lymphoma of colon presenting with multiple polypoid lesions. PMID- 11007253 TI - New mutation in the hMSH2 gene in a Spanish Muir-Torre syndrome. PMID- 11007254 TI - Whipple's disease: acquired resistance to trimethoprim-sulfamethoxazole. PMID- 11007255 TI - Autoimmune pancreatitis and hepatitis: an uncommon association. PMID- 11007256 TI - Prevention of pancreatitis by weekly amylase assay in patients with Crohn's disease treated with azathioprine. PMID- 11007257 TI - Frequency of thrombophilic abnormalities in Turkish children with inflammatory bowel disease. PMID- 11007258 TI - Severe anaphylactic reaction to infliximab: report of a case. PMID- 11007259 TI - Primary biliary cirrhosis exacerbated by a course of acute hepatitis C and subsequent interferon therapy. PMID- 11007260 TI - Endoscopic stent pretzel. PMID- 11007261 TI - Is the risk alkaline reflux a risk factor for laryngeal lesions? PMID- 11007262 TI - Biochemical and virological changes in serum of nonresponder or relapser chronic hepatitis C patients retreated with interferon and ribavirin followed by interferon alone. PMID- 11007263 TI - An unusual cause of massive gastric bleeding in a young patient. PMID- 11007264 TI - Percutaneous microwave coagulation therapy for superficial hepatocellular carcinoma on the liver surface. PMID- 11007265 TI - Successful treatment of lymphoid follicular proctitis with sulfasalazine suppositories. PMID- 11007266 TI - Artificial ascites method: percutaneous treatments for hepatocellular carcinoma located just beneath the diaphragm. PMID- 11007267 TI - Transitory ischemic attack and acute hepatitis C. PMID- 11007268 TI - Selective and facile cyclization of N-chloroacetylated peptides from the C4 domain of HIV Gp120 in LiCl/DMF solvent systems. AB - Lithium salts have been reported to mediate the solubilization of peptides in organic solvents in 1989 (Seebach, D., Thaler, A. & Beck, A. K. Helv. Chim. Acta 1989; 72, 857-867). The use of Li salts in an organic solvent to influence cyclization of a reactive peptide that only polymerizes in an aqueous solvent, has not been reported. Here, the selective and facile cyclization of N chloroacetylated, C-cysteine amide peptides from the C4 domain of HIV-1 gp120 in LiCl/DMF solvent systems is demonstrated. The addition of stoichiometric amounts of Tris base to 1 mg/mL peptide in LiCl/DMF solutions was sufficient to drive the cyclization to completion within 3 h at ambient temperatures. Cyclic peptides were the only detectable reaction products and these were confirmed using reversed-phase HPLC and mass spectrometric analyses of the final products. In aqueous solutions at pH 7.4, only polymers were obtained as judged by HPLC and SDS-PAGE. The method of using Li salts in an organic solvent to enhance the cyclization of unprotected amphipathic peptides may be useful in many situations beyond those described here. PMID- 11007269 TI - Influence of the hydrophilic face on the folding ability and stability of alpha helix bundles: relevance to the peptide catalytic activity. AB - Although not the sole feature responsible, the packing of amino acid side chains in the interior of proteins is known to contribute to protein conformational specificity. While a number of amphipathic peptide sequences with optimized hydrophobic domains has been designed to fold into a desired aggregation state, the contribution of the amino acids located on the hydrophilic side of such peptides to the final packing has not been investigated thoroughly. A set of self aggregating 18-mer peptides designed previously to adopt a high level of alpha helical conformation in benign buffer is used here to evaluate the effect of the nature of the amino acids located on the hydrophilic face on the packing of a four alpha-helical bundle. These peptides differ from one another by only one to four amino acid mutations on the hydrophilic face of the helix and share the same hydrophobic core. The secondary and tertiary structures in the presence or absence of denaturants were determined by circular dichroism in the far- and near UV regions, fluorescence and nuclear magnetic resonance spectroscopy. Significant differences in folding ability, as well as chemical and thermal stabilities, were found between the peptides studied. In particular, surface salt bridges may form which would increase both the stability and extent of the tertiary structure of the peptides. The structural behavior of the peptides may be related to their ability to catalyze the decarboxylation of oxaloacetate, with peptides that have a well-defined tertiary structure acting as true catalysts. PMID- 11007270 TI - Solution conformational study of Scyliorhinin I analogues with conformational constraints by two-dimensional NMR and theoretical conformational analysis. AB - Two analogues of Scyliorhinin I (Scyl), a tachykinin with N-MeLeu in position 8 and a 1,5-disubstituted tetrazole ring between positions 7 and 8, introduced in order to generate local conformational constraints, were synthesized using the solid-phase method. Conformational studies in water and DMSO-d6 were performed on these peptides using a combination of the two-dimensional NMR technique and theoretical conformational analysis. The algorithm of conformational search consisted of the following three stages: (i) extensive global conformational analysis in order to find all low-energy conformations; (ii) calculation of the NOE effects and vicinal coupling constants for each of the low energy conformations; (iii) determining the statistical weights of these conformations by means of a nonlinear least-squares procedure, in order to obtain the best fit of the averaged simulated spectrum to the experimental one. In both solvents the three-dimensional structure of the analogues studied can be interpreted only in terms of an ensemble of multiple conformations. For [MeLeu8]Scyl, the C-terminal 6-10 fragment adopts more rigid structure than the N-terminal one. In the case of the analogue with the tetrazole ring in DMSO-d6 the three-dimensional structure is characterized by two dominant conformers with similar geometry of their backbones. They superimpose especially well (RMSD = 0.28 A) in the 6-9 fragments. All conformers calculated in both solvents superimpose in their C-terminal fragments much better than those of the first analogue. The results obtained indicate that the introduction of the tetrazole ring into the Scyl molecule rigidifies its structure significantly more than that of MeLeu. PMID- 11007271 TI - Structure-bioactivity of C-terminal pentapeptide of osteogenic growth peptide [OGP(10-14)]. AB - The amino acid sequence of osteogenic growth peptide (OGP) consists of 14 residues identical to the C-terminal tail of histone H4. Native and synthetic OGP are mitogenic to osteoblastic and fibroblastic cells and enhance osteogenesis and hematopoiesis in vivo. The C-terminal truncated pentapeptide of OGP, H-Tyr-Gly Phe-Gly-Gly-OH [OGP(10-14)], is a naturally occurring osteoblastic mitogen, equipotent to OGP. The present study assesses the role of individual amino acid residues and side chains in the OGP(10-14) mitogenic activity which showed a very high correlation between osteoblastic and fibroblastic cell cultures. Truncation of either Tyr10 or its replacement by Ala or D-Ala resulted in substantial, but not complete, loss of activity. Nevertheless, only a small loss of activity was observed following removal of the Tyr10 amino group. No further loss occurred consequent to the monoiodination of desaminoTyr10 on meta-position. However, a marked decrease in proliferative activity followed removal of the Tyr10 phenolic or the Phe12 aromatic group. Loss of activity of a similar magnitude also occurred subsequent to replacing Gly11 with L- or D-Ala. Approximately 50% loss of mitogenic activity occurred subsequent to truncation of Gly14 or blocking the C-terminal group as the methyl ester. All other modifications of the C-terminus and L- or D-Ala substitution of Gly13 resulted in 70-97% decrease in activity. Collectively, these data suggest that the integrity of the pharmacophores presented by Tyr and Phe side chains, as well as the Gly residues at the C terminus, are important for optimal bioactivity of OGP(10-14). PMID- 11007272 TI - Chaperone-like activity of a synthetic peptide toward oxidized gamma-crystallin. AB - alphaA-Crystallin can function like a molecular chaperone. We recently reported that the alphaA-crystallin sequence, KFVIFLDVKHFSPEDLTVK (peptide-1, residues 70 88) by itself possesses chaperone-like (anti-aggregating) activity during a thermal denaturation assay. Based on the above data we proposed that the peptide 1 sequence was the functional site in alphaA-crystallin. In this study we investigated the specificity of peptide-1 against gamma-crystallin aggregation in the presence of H2O2 and CuSO4. Peptide-1 was able to completely protect against the oxidation-induced aggregation of gamma-crystallin. Removal of N-terminal Lys or the replacement of Lys with Asp (DFVIFLDVKHFSPEDLTVK, peptide-2) did not alter the anti-aggregation property of peptide-1. However, deletion of KF residues from the N-terminus of peptide-1 resulted in a significant loss of its anti aggregation property. Bio-gel P-30 size-exclusion chromatography of gamma crystallin incubated with peptide-2 under oxidative conditions revealed that a major portion of the peptide elutes in the void volume region along with gamma crystallin, suggesting the binding of the peptide to the protein. Peptide-1 and 2 were also able to prevent the UV-induced aggregation of gamma-crystallin. These data indicate that the same amino acid sequence in alphaA-crystallin is likely to be responsible for suppressing the heat-denatured, oxidatively modified and UV induced aggregation of proteins. PMID- 11007273 TI - Reactivity toward deamidation of asparagine residues in beta-turn structures. AB - Mimetics of beta-turn structures in proteins have been used to calibrate the relative reactivities toward deamidation of asparagine residues in the two central positions of a beta-turn and in a random coil. N-Acetyl-Asn-Gly-6 aminocaproic acid, an acyclic analog of a beta-turn mimic undergoes deamidation of the asparaginyl residue through a succinimide intermediate to generate N acetyl-Asp-N-Gly-6-aminocaproic acid (6-aminocaproic acid, hereafter Aca) and N acetyl-L-iso-aspartyl (isoAsp)-Gly-Aca (pH 8.8, 37 degrees C) approximately 3 fold faster than does the cyclic beta-turn mimic cyclo-[L-Asn-Gly-Aca] with asparagine at position 2 of the beta-turn. The latter compound, in turn, undergoes deamidation approximately 30-fold faster than its positional isomer cyclo-[Gly-Asn-Aca] with asparagine at position 3 of the beta-turn. Both cyclic peptides assume predominantly beta-turn structures in solution, as demonstrated by NMR and circular dichroism characterization. The open-chain compound and its isomer N-acetyl-Gly-Asn-Aca assume predominantly random coil structures. The latter isomer undergoes deamidation 2-fold slower than the former. Thus the order of reactivity toward deamidation is: asparagine in a random coil approximately 3x(asparagine) in position 2 of a beta-turn approximately 30x (asparagine) in position 3 of a beta-turn. PMID- 11007274 TI - Structural study of the sodium channel inactivation gate peptide including an isoleucine-phenylalanine-methionine motif and its analogous peptide (phenylalanine/glutamine) in trifluoroethanol solutions and SDS micelles. AB - In order to gain insight into the gating mechanisms of Na+ channels, in particular their inactivation mechanisms, we studied the structures of the Na+ channel inactivation gate related peptide which includes the IFM (Ile-Phe-Met) motif (Ac-KKKFGGQDIFMTEEQKK-NH2; K1480-K1496 in rat brain type-IIA Na+ channels, MP-3A) and its F/Q(Gln) substituted one (MP-4A) in trifluoroethanol (TFE) solutions and sodium dodecyl sulfate (SDS) micelles using circular dichroism (CD) and 1H-NMR spectroscopies. Based on observed nuclear Overhauser effect constraints, three-dimensional structures of MP-3A and MP-4A were determined using simulated annealing molecular dynamics/ energy minimization calculations. In TFE solutions, no appreciable differences in the structure were observed using either CD or NMR spectra. In SDS micelles, however, the two peptides exhibited definitely different structures from each other. It was found that in MP-3A, residues 11488 and T1491 were spatially proximate with each other owing to hydrogen bonding between the amide proton of 11488 and the hydroxyl oxygen atom of T1491, whereas in MP-4A, F/Q substitution separated them owing to conformational changes. The solvent-accessible surfaces calculated for the structures of MP-3A and MP-4A showed that the former has a smoother interaction surface to the hydrophobic docking site than the latter. In conclusion, the conformational changes, as well as decreased hydrophobicity around the IFM motif owing to the F/Q mutation, may be one reason why F1489Q mutated channels cannot inactivate almost completely. PMID- 11007275 TI - TBT--towards a better way to regulate pollutants. PMID- 11007276 TI - Cost-benefit analysis of the use of TBT: the case for a treatment approach. AB - The current climate of hostility towards the use of tributyltin (TBT) as an active ingredient in ship anti-fouling paint appears to be based on a very biased assessment of its environmental impact. While many national and international regulatory agencies are moving towards further restriction, and a complete ban is under active discussion, a number of factors appear to have been ignored. The economic impact of a ban on TBT when no adequate substitute exists could be substantial. Environmentally, consequences would include a substantial increase in the consumption of fossil fuel, with corresponding increases in carbon dioxide and sulphur dioxide emissions; the construction of more vessels; the transfer of ship-building, ship-repairing and ship-breaking activities from well-regulated to unregulated or under-regulated areas in the developing world; and a shift from sea transport to less environmentally acceptable forms of transport. Experience in Europe and other parts of the developed world shows that existing restrictions, where they are properly enforced, are probably adequate to alleviate the environmental damage associated with TBT. Some existing legislation acts to inhibit the search for effective substitutes. The environmental benefits of TBT have been ignored. Little thought has been given to a technical, rather than a legislative solution to controlling TBT inputs to the environment. A method is described for treating TBT-contaminated wastewaters, which has been successfully tested in prototype at full scale. Legislative measures against TBT will do nothing to address the problem of the existing backlog of contaminated material, nor even to permit the IMO proposal for the removal of TBT from all ships by 2008 to be successfully concluded in an environmentally safe manner, since no provision has been made for the disposal of the existing TBT; most probably it will be dumped in environmentally sensitive, unregulated areas in the developing world. PMID- 11007278 TI - The use of imposex to assess tributyltin contamination in coastal waters and open seas. AB - Imposex in muricid gastropods has been used to monitor tributyltin (TBT) contamination worldwide. Pollution was severe during the 1980s but regulations prohibiting the use of TBT-based antifoulants on vessels < 25 m in length have been highly effective in reducing TBT levels in coastal waters. Large vessels are still sources of TBT and major ports, especially those with dry docking and repair facilities, continue to be hot-spots of contamination. Measures of imposex suggest that severe pollution is normally localised to within a few kilometres of them. Mild imposex, possibly as a result of TBT pollution (although other causes have not been ruled out), has been described in whelks Buccinum undatum from offshore waters of the southern and central North Sea. However, the species is still abundant there and the occurrence of imposex does not seem to have affected its breeding performance. Imposex was mild or absent in populations of dogwhelks from open oceanic sites around the north Atlantic Ocean, suggesting that the oceans are free of contamination at biologically significant levels. PMID- 11007277 TI - A review of organotin regulatory strategies, pending actions, related costs and benefits. AB - Achieving consensus on equitable and effective national and global regulation(s) for the use of organotins as biocides in antifouling boat bottom paints has proven to be very complex and difficult for a variety of reasons as discussed in this paper. There appears to be broad agreement among stakeholders about the effectiveness of tributyltin (TBT) in antifouling paints. A draft Assembly Resolution prepared by the Marine Environmental Protection Committee (MEPC) of the International Maritime Organization (IMO) to propose a global ban on the use of organotins in antifouling paints was approved by the IMO at its 21st regular session (November 1999). In approving the Resolution, the Assembly agreed that a legally binding instrument (global convention--an international treaty) be developed by the Marine Environmental Protection Committee that should ensure by 1 January 2003, a ban on the application of tributyltin (TBT)-based antifouling paints; and 1 January 2008 as the last date for having TBT-based antifouling paint on a vessel. The Assembly also agreed that a diplomatic conference be held in 2001 to consider adoption of the international legal instrument. Monitoring, policing, enforcement, fines and record-keeping are yet to be defined. In addition, the MEPC has also proposed that IMO promotes the use of environmentally safe anti-fouling technologies to replace TBT. Existing national regulations in the US and Europe have: (1) restricted the use of TBT in antifouling boat bottom paints by vessel size (less than 25 m in length), thus eliminating TBT from the smaller and recreational vessels that exist in shallow coastal waters where the impacted oysters species grow; (2) restricted the release rates of TBT from co polymer paints; and (3) eliminated the use of free TBT in paints. The present movement toward a global ban suggests that the above regulatory approach has not been sufficient in some countries. Advocates of the ban cite international findings of: (1) higher levels of TBT in surface waters of ports and open waters; (2) imposex still occurring and affecting a larger number of snail species; (3) TBT bioaccumulation in selected fisheries; and (4) the availability of 'comparable' alternatives (to TBT) with less environmental impact. The global ban has been absent of a policy debate on the: (1) lack of 'acceptable and approved' alternatives in many nations; (2) appreciation of market forces in nations without TBT regulations; (3) full consideration of the economic benefits from the use of TBT; (4) 'acceptance' of environmental impacts in marinas, ports and harbors; and (5) realization of the 'real' time period required by ships for antifoulant protection (is 5-7 years necessary or desirable?). Estimates of fuel savings range from $500 million to one billion. In assessing the environmental impact from TBT, there are two sources: the shipyard painting vessels and the painted vessel itself. Today vessels can be painted with regulated or banned antifouling materials by boatyards in a country that does not have TBT regulations and subsequently travel in international and regulated national waters and thus bringing the impact back to the country which was trying to prevent it. Worse, local and national regulations for TBT have proven to be the antithesis of the popular environmental cliche--'Think Globally and Act Locally.' Legislative policies enacted by 'regulated' countries to regulate the use of TBT to protect (their) local marine resources have subsequently had far reaching environmental and economic impacts which have in essence transferred TBT contamination to those countries least able to deal with it. Market forces are selective for cheap labor and cheap environments. 'Unregulated' countries have unknowingly accepted the environmental and human health risks to gain the economic benefits from painting TBT on ships. (ABSTRACT TRUNCATED) PMID- 11007279 TI - Analytical aspects of the gas chromatographic determination of tributyltin and metabolites in environmental samples. AB - Tributyltin and its metabolites were determined in environmental samples by gas chromatography using a flame photometric detection method. The analytical method involved the propylation of the organotins and the use of a recovery standard to correct for inefficient recovery. A detection limit of 5 ng Sn l(-1) for seawater, and 5 ng Sn g(-1) for sediments, was obtained when the detector was equipped with an optical filter; however, this concentration limit, which is higher than the environmental quality standard for TBT, could be improved by a factor of 100 on removing the filter. The limitations imposed by the choice of recovery and internal standards were discussed and it was shown that the possible presence of Sn(II) had no effect on the total extractable inorganic tin, as determined by the analytical protocol. The identification of unexpected mixed methylbutyltins in TBT-contaminated sediments was described, and the implications of the existence of such compounds were discussed. PMID- 11007280 TI - Evaluation of environmental levels and biological impact of TBT in Malta (central Mediterranean). AB - Chemical monitoring for butyltins in bulk seawater, surface microlayer and superficial sediments determined that the two main local sources of marine contamination by tributyltin (TBT) compounds in Malta are ship-repairing dockyards and marinas. In bulk seawater, TBT values ranged from below the detection limit of 5 ng Sn l(-1) to 300 ng Sn l(-1); in sediments derived from the most polluted areas, TBT concentrations as high as 1500 ng Sn g(-1) were measured. At TBT levels found in local harbours, several sublethal biological responses are possible and were observed, including a significant reduction in MFO enzyme system activities of fish; digestive cell atrophy in the oyster Ostrea edulis; and induction of imposex in the snail Hexaplex trunculus. The latter two responses are evident at TBT concentrations below the environmental quality standard (20 ng TBT l(-1)). The ecological implications of these results are discussed and it is concluded that exposure of marine organisms to TBT in local harbours may be expected to lead to a reduction in the long-term survival of a number of such organisms. For these reasons, both the use of TBT-based antifouling paints for pleasure boats as well as drydock practices need to be controlled by appropriate regulations and their enforcement. PMID- 11007281 TI - Environmental impact of TBT: the French experience. AB - The deleterious effects of TBT released by antifouling paints were first documented in Arcachon Bay (France) at the end of the 1970s. As a result of the high level of water contamination, the production of Pacific oysters was severely affected by a complete lack of reproduction and the appearance of calcification anomalies which were responsible for a strong decline in the marketable value of the remaining stock. Regulations that had been successively adopted from January 1982 had positive effects on the recovery of the oyster grounds by reducing ambient water concentrations. Most antifouling paints act by releasing toxicants into the water which avoid the fixation of the organisms responsible for fouling. The use of these paints results in significant inputs of biocides in the waters. The harmful effects on non-target organisms were first evidenced in the bay of Arcachon (France) where the most acute disturbances were recorded following tributyltin (TBT) contamination. PMID- 11007282 TI - Sea-nine antifoulant: an environmentally acceptable alternative to organotin antifoulants. AB - This article reviews previously reported data on the performance, environmental fate, and environmental modeling of Sea-Nine 211 antifoulant (4,5-dichloro-2-n octyl-4-isothiazolin-3-one). Since organotins are an industry standard, the environmental fate and environmental modeling results of tributyltin (TBT) are compared to those of the Sea-Nine antifoulant for reference. Laboratory and field tests results have shown Sea-Nine antifoulant to be highly effective. Five years of commercial use has confirmed this. Sea-Nine antifoulant and TBT were compared in an environmental risk assessment to predict their effects on the environment. Sea-Nine antifoulant was degraded rapidly in the environment by microorganisms. Its half-life in aerobic and anaerobic microcosm studies was less than 1 h. TBT degraded slowly under aerobic and anaerobic conditions with half-lives ranging from 6 to 9 months. The degradation products of Sea-Nine antifoulant were ring opened compounds with greatly reduced toxicity. TBT degraded to dibutyltin species, which were still toxic and persistent in the environment. Bioaccumulation studies in fish showed essentially no bioaccumulation of the Sea Nine biocide. The bioaccumulation of TBT was significant, with bioconcentration factors as high as 10000. The Sea-Nine antifoulant showed no chronic or reproductive toxicity to marine species, while TBT showed a wide range of effects on growth, development, and reproduction at levels as low as 2 parts per trillion (ppt). Computer modeling using the Exposure Analysis Modeling System (EXAMS) predicted maximum concentrations of Sea-Nine biocide of up to 10 ppt, far below the maximum acceptable environmental concentration (MAEC) of 630 ppt. The maximum predicted concentrations of TBT were as high as 345 ppt, far above the UK Environmental Standard in seawater of 2 ppt. PMID- 11007283 TI - The second order effects on commercial shipping of restrictions on the use of TBT. AB - Increased hull roughness reduces vessel speed and increases fuel consumption. These are first order effects of restricting or prohibiting the use of antifouling paints. Reduced transport capacity and increased capital costs are frequently recognised as second order effects of such reductions in fleet efficiency. This paper focuses on other second order effects of prohibiting or restricting the use of TBT-based antifoulings. The potential cost increases will affect high-speed vessels most severely. These vessels are the segment of the shipping industry that is most relevant to the political drive in Europe towards shifting the balance of transport of goods towards sea rather than road transport for environmental reasons. Any decision to restrict or prohibit these paints would tend also to induce a shift from local to global environmental problems, which would be more difficult to manage. PMID- 11007284 TI - Microbial interactions with tributyltin compounds: detoxification, accumulation, and environmental fate. AB - While inorganic forms of tin are of relatively low toxicity towards microorganisms, the more lipid-soluble organotins can be highly toxic. Generally, trisubstituted (R3SnX) organotins are more toxic than di- (R2SnX2) and monosubstituted (RSnX3) compounds; the anion (X) apparently having little influence on toxicity. However, many microorganisms exhibit resistance to organotins, a phenomenon of relevance to the environmental cycling of organotins and also to novel biological methods of treatment. Organotin degradation can involve the sequential removal of organic moieties to yield less toxic derivatives, e.g. debutylation of tributyltin compounds to di- and monobutylins. Such degradation is known to take place in bacteria, algae and fungi, and this provides one route for detoxification. In addition, microorganisms are capable of accumulating tributyltin compounds, and this is another mechanism of removal from solution. The high lipid solubility of organotins ensures cell penetration and association with intracellular sites, while cell wall components also play an important role. Of the fungal wall components, melanin pigments are capable of TBT binding, and the addition of melanin to growing cultures can remove toxicity; melanised strains are also more sensitive than albino strains of the same species. To date, little attention has been paid to the biotechnological exploitation of these interactions for the degradation of tributyltin or its removal from solution. This paper describes some interactions of microorganisms (bacteria, cyanobacteria, microalgae, and fungi) with tributyltin compounds, with particular reference to toxicity, bioaccumulation and detoxification. Such processes should receive due consideration in any environmental management programme. PMID- 11007285 TI - An alternative approach to antifouling based on analogues of natural processes. AB - A number of marine organisms are able to resist fouling pressure and remain essentially free of fouling. Some organisms are totally devoid of even the first stages of biofilm formation involving bacteria and microalgae. A key feature in recent research has been the realisation that previous low adherence technology is an insufficient technical solution and that natural models, based on marine and other organisms, incorporate other passive techniques for fouling resistance. These characteristics may be incorporated into physical analogues of the natural processes. This paper describes ways of producing physical analogues of some such characteristics, the application of such techniques to surfaces in the marine environment and the environmental impact. The paper includes some results of recent trials and a cost comparison. PMID- 11007286 TI - Leaching, runoff and speciation of arsenic in a laboratory mesocosm. AB - Leaching and runoff of arsenic (As) from the contaminated soil of an old wood impregnation plant, and fate in a recipient freshwater ecosystem, was studied in soil-water-sediment mesocosms in laboratory (0.9 m3; total water volume 200 l). During the 4-month experiment total leaching and discharge of As from regularly irrigated soil was approximately 40 mg, i.e. approximately 0.6% of total initial As content in the soil. Of the total As load, 7.5% remained in the water; 44% settled down to the shallow (water depth 5-30 cm) sediment zone; and 48.5% to the deeper (water depth 80 cm) sediment zone. The different arsenic species; arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid (MMAA) and dimethylarsinic acid (DMAA), were analysed from irrigation and discharge water; mesocosm pool water; and sediment pore water using ion chromatography-inductively coupled plasma-mass spectrometry (IC-ICP-MS). The total amounts of arsenic in soil, water and sediment were determined by ICP-MS. Arsenic was leached out from the soil as As(V). In mesocosm water As(V) was the predominant dissolved species, but DMAA and particle bound species, were also detected. In shallow sediment, As(V) was the most abundant species together with some DMAA, whereas in deep sediment As(III) was the dominant species. PMID- 11007287 TI - Selective chemical extraction and grainsize normalisation for environmental assessment of anoxic sediments: validation of an integrated procedure. AB - An integrated selective extraction and size normalisation procedure for use in metal partitioning and diagenetic studies of anoxic sediments and soils is presented. Data obtained by this procedure can readily be combined with other primary data (e.g. sulfur concentrations, carbonate concentrations, cation exchange capacity, etc.) and derived parameters (e.g. degrees of pyritisation and sulfidisation) that enhance interpretation of the behaviour of trace metals in anoxic sediments. Achieving size normalisation during extraction, allows direct comparison of sediments from dissimilar sedimentary environments, and simplifies assessment of the processes that determine whether a sediment is a source of or a sink for trace metals. Aspects of a study of trace metals in sediments from the Brisbane River estuary, Australia, are used to illustrate applications of the integrated procedure. PMID- 11007288 TI - Long-term monitoring of arsenic and selenium species in contaminated groundwaters by HPLC and HG-AAS. AB - The long-term concentration and distribution of species of arsenic and selenium in contaminated groundwaters from Kelheim was monitored. Most of the groundwater wells contained elevated concentrations of iron, manganese and sulfur. Arsenic (III), arsenic (V), selenium (IV) and selenium (VI) were separated using high performance liquid chromatography (HPLC) based on phosphate buffers and collected in fractions. Due to the complex matrix, the fractions were analyzed element specifically by hydride-generating atomic absorption spectrometry (HG-AAS). The combination of HPLC and HG-AAS was selected due to the authors' intention of developing an easy-to-handle, but nonetheless reliable, method suitable for the long-term monitoring of species distribution in an almost routine way, and taking account of the threshold values of 10 microg/l for each element, indicated by German drinking water regulations. To enhance the reliability of the method, analytical quality control experiments were carried out. When applied to groundwater wells from Kelheim (Germany) they revealed that arsenic (V) and selenium (VI) were the dominating species. The presence of arsenic (III) and selenium (IV) was assumed to be supported by organic matter. PMID- 11007289 TI - Speciation of aluminium in forest soil extracts by size exclusion chromatography with UV and ICP-AES detection and cation exchange fast protein liquid chromatography with ETAAS detection. AB - Aluminium speciation was studied in forest soil extracts by size exclusion chromatography (SE) with UV and inductively coupled plasma-atomic emission spectrometric (ICP-AES) detection and cation exchange fast protein liquid chromatography (FPLC) with ETAAS detection. Size exclusion chromatography was performed on a Superdex HR75 10/30 column. Isocratic clution with 0.15 mol dm(-3) NaCl in TRIS-HCl buffer (pH = 5.5) was applied over 100 min at a flow rate of 0.35 cm(3) min(-1). The chromatographic run was followed at 278 nm and separated Al species also determined 'off line' in 0.875 cm3 fractions by ICP-AES. The analytical procedure enabled speciation of high molecular weight Al complexes. Cation exchange FPLC was performed on a Mono S HR 5/5 column. Aqueous 8 mol dm( 3) NH4NO3 linear gradient elution was applied over 10 min at a flow rate of 1 cm3 min(-1). Separated Al species were collected in 0.5 cm3 fractions and Al determined 'off line' by ETAAS. The analytical procedure enabled speciation of some positively charged monomeric Al species. Negatively charged species were eluted with the solvent front. The combination of the two analytical techniques was successfully employed in speciation of Al in forest soil extracts. Water was used as an extracting solution. It was found experimentally that 80-95% of Al in aqueous extracts of forest soils exists in monomeric Al forms. Water soluble Al (30-40%) is bound to high molecular weight complexes with humic substances. The remaining monomeric Al in the low molecular weight fraction exists as AIF2+, Al oxalate and Al-citrate species. PMID- 11007290 TI - Phosphorus cycling through phosphine in paddy fields. AB - Phosphine emission fluxes from paddy fields, phosphine ambient levels in air, and the vertical profile of matrix-bound phosphine in soil have been measured throughout the growing season of rice in Beijing, China. It was found that both the seasonal and diurnal emission fluxes and ambient levels fluctuate significantly. During the drainage period, phosphine released from the soil with the highest diurnal average flux on the first period of drainage (approx. 17.7 ng m(-2) h(-1)), whereas its highest ambient level (approx. 250 ng m(-3)) occurred at 06.00 h. During the flooded period, phosphine emission was low, and the peaks of phosphine emissions occurred at midnight. The average flux of PH3 emission for the whole season was found to be approximately 1.78 ng m(-2) h(-1). The mass fraction of matrix-bound phosphine is approximately 0.18 approximately 1.42 x 10( 7) (m/m) part of organic phosphorus or 3.4 approximately 9.2 x 10(-9) (m/m) part of total phosphorus in paddy soil. The amount of phosphine emitted to the atmosphere was only a small fraction of the phosphine that remained in the soil in the matrix-bound form. Soil serves both as the source and the sink of PH3. PMID- 11007291 TI - Nutrient elements of commercial tea from Nigeria by an instrumental neutron activation analysis technique. AB - A prototype miniature neutron source reactor (MNSR) with a thermal neutron flux of 3.0 x 10(11) n cm(-2) s(-1) has been used to determine the concentrations of some nutrient elements leading to short-lived activation products in commercial tea leaf samples from Nigeria. A total of eight elements Al, Ca, Cl, Cu, K, Mg, Mn and Na, that can be routinely used for quality control purposes, were analyzed in this study. Two biological reference materials, tomato leaves (NIST-1573) and citrus leaves (NIST-1572) were used as the standard and quality control materials, respectively. The analytical results show that the average concentrations of Al, Ca, Cl, Cu, K, Mg, Mn and Na in Nigerian tea are slightly higher when compared with a Chinese herbal tea analyzed in this study. The concentration ratios of K/Ca were found to be high in all the samples analyzed suggesting cultivation in potash-rich soils. PMID- 11007292 TI - Radiological consequences of Chernobyl accident: UN Scientific Committee on Effects of Atomic Radiation confirms earlier IAEA assessments. PMID- 11007293 TI - Navigation in biological and artificial systems. PMID- 11007294 TI - How honeybees make grazing landings on flat surfaces. AB - Freely flying bees were filmed as they landed on a flat, horizontal surface, to investigate the underlying visuomotor control strategies. The results reveal that (1) landing bees approach the surface at a relatively shallow descent angle; (2) they tend to hold the angular velocity of the image of the surface constant as they approach it; and (3) the instantaneous speed of descent is proportional to the instantaneous forward speed. These characteristics reflect a surprisingly simple and effective strategy for achieving a smooth landing, by which the forward and descent speeds are automatically reduced as the surface is approached and are both close to zero at touchdown. No explicit knowledge of flight speed or height above the ground is necessary. A model of the control scheme is developed and its predictions are verified. It is also shown that, during landing, the bee decelerates continuously and in such a way as to keep the projected time to touchdown constant as the surface is approached. The feasibility of this landing strategy is demonstrated by implementation in a robotic gantry equipped with vision. PMID- 11007295 TI - Wide-field, motion-sensitive neurons and matched filters for optic flow fields. AB - The receptive field organization of a class of visual interneurons in the fly brain (vertical system, or VS neurons) shows a striking similarity to certain self-motion-induced optic flow fields. The present study compares the measured motion sensitivities of the VS neurons (Krapp et al. 1998) to a matched filter model for optic flow fields generated by rotation or translation. The model minimizes the variance of the filter output caused by noise and distance variability between different scenes. To that end, prior knowledge about distance and self-motion statistics is incorporated in the form of a "world model". We show that a special case of the matched filter model is able to predict the local motion sensitivities observed in some VS neurons. This suggests that their receptive field organization enables the VS neurons to maintain a consistent output when the same type of self-motion occurs in different situations. PMID- 11007296 TI - Combined space-variant maps for optical-flow-based navigation. AB - A robot navigating in an unstructured environment needs to avoid obstacles in its way and determine free spaces through which it can safely pass. We present here a set of optical-flow-based behaviors that allow a robot moving on a ground plane to perform these tasks. The behaviors operate on a purposive representation of the environment called the "virtual corridor" which is computed as follows: the images captured by a forward-facing camera rigidly attached to the robot are first remapped using a space-variant transformation. Then, optical flow is computed from the remapped image stream. Finally, the virtual corridor is extracted from the optical flow by applying simple but robust statistics. The introduction of a space-variant image preprocessing stage is inspired by biological sensory processing, where the projection and remapping of a sensory input field onto higher-level cortical areas represents a central processing mechanism. Such transformations lead to a significant data reduction, making real time execution possible. Additionally, they serve to "re-present" the sensory data in terms of ecologically relevant features, thereby simplifying the interpretation by subsequent processing stages. In accordance with these biological principles we have designed a space-variant image transformation, called the polar sector map, which is ideally suited to the navigational task. We have validated our design with simulations in synthetic environments and in experiments with real robots. PMID- 11007297 TI - The turn-back-and-look behaviour: bee versus robot. AB - Honeybees and social wasps departing from a novel food source perform stereotype flight manoeuvres, termed the turn-back-and-look behaviour (TBL). Based on results of behavioural studies, it is proposed that the image motion generated by the TBL provides the insect with information about the three-dimensional structure of the goal's surroundings, thus enabling it to select reliable landmarks that will guide it to the goal upon return. The colour, shape, and size of landmarks, on the other hand, are learned mainly during arrival at the food source. However, when bees are prevented from learning these cues on arrival, they learn them during the TBL, despite the fact that this performance does not require the use of image motion. A recently developed model shows that landmark learning can indeed be accomplished during the TBL by exploiting cues others than image motion. A mobile robot equipped with the appropriate software selects, during the TBL, reliable marks and returns to the site of departure from different locations by accomplishing image matching along a two-dimensional vector field. PMID- 11007298 TI - Insect visual homing strategies in a robot with analog processing. AB - The visual homing abilities of insects can be explained by the snapshot hypothesis. It asserts that an animal is guided to a previously visited location by comparing the current view with a snapshot taken at that location. The average landmark vector (ALV) model is a parsimonious navigation model based on the snapshot hypothesis. According to this model, the target location is unambiguously characterized by a signature vector extracted from the snapshot image. This article provides threefold support for the ALV model by synthetic modeling. First, it was shown that a mobile robot using the ALV model returns to the target location with only small position errors. Second, the behavior of the robot resembled the behavior of bees in some experiments. And third, the ALV model was implemented on the robot in analog hardware. This adds validity to the ALV model, since analog electronic circuits share a number of information processing principles with biological nervous systems; the analog implementation therefore provides suggestions for how visual homing abilities might be implemented in the insect's brain. PMID- 11007299 TI - How do insects use path integration for their navigation? AB - We combine experimental findings on ants and bees, and build on earlier models, to give an account of how these insects navigate using path integration, and how path integration interacts with other modes of navigation. At the core of path integration is an accumulator. This is set to an initial state at the nest and is updated as the insect moves so that it always reports the insect's current position relative to the nest. Navigation that uses path integration requires, in addition, a way of storing states of the accumulator at significant places for subsequent recall as goals, and a means of computing the direction to such goals. We discuss three models of how path integration might be used for this process, which we call vector navigation. Vector navigation is the principal means of navigating over unfamiliar terrain, or when landmarks are unavailable. Under other conditions, insects often navigate by landmarks, and ignore the output of the vector navigation system. Landmark navigation does not interfere with the updating of the accumulator. There is an interesting symmetry in the use of landmarks and path integration. In the short term, vector navigation can be independent of landmarks, and landmark navigation needs no assistance from path integration. In the longer term, visual landmarks help keep path vector navigation calibrated, and the learning of visual landmarks is guided by path integration. PMID- 11007300 TI - Triple-loop model of path control by head direction and place cells. AB - Arthropods as well as mammals are able to return straight home after a random search excursion under conditions that are designed to exclude all external cues. After a brief clarification of the terminology, two principal systems of information processing that can achieve this performance are introduced and analysed: Polar versus Cartesian path integration. The different demands and achievements of the two systems are confronted with neurophysiological findings on the functioning of the hippocampus, and with a recent comprehensive model of how the hippocampal place cells perform path integration. To connect the neurophysiological findings with the behavior of the animal, a new model is developed. It achieves three functionally diverse performances: maintenance and control of a compass direction, navigation by path integration, and formation of goals by connecting non-spatial features with their location. This is done by three interconnected feedback loops, set by a common reference variable. Their information-processing structure enables the animal not only to home but also to go straight from any stored goal to any other, without explicit representation of the distance between them, and without a topological arrangement of the store. The model explains behaviors not yet understood and predicts still undiscovered performances. Because it allows the isolation of orienting from storing functions yet also shows how they can be connected, the model may help to reconcile conflicting views on the function of the hippocampus. PMID- 11007301 TI - Animat navigation using a cognitive graph. AB - This article describes a computational model of the hippocampus that makes it possible for a simulated rat to navigate in a continuous environment containing obstacles. This model views the hippocampus as a "cognitive graph", that is, a hetero-associative network that learns temporal sequences of visited places and stores a topological representation of the environment. Calling upon place cells, head direction cells, and "goal cells", it suggests a biologically plausible way of exploiting such a spatial representation for navigation that does not require complicated graph-search algorithms. Moreover, it permits "latent learning" during exploration, that is, the building of a spatial representation without the need of any reinforcement. When the rat occasionally discovers some rewarding place it may wish to rejoin subsequently, it simply records within its cognitive graph, through a series of goal and sub-goal cells, the direction in which to move from any given start place. Accordingly, the model implements a simple "place-recognition-triggered response" navigation strategy. Two implementations of place cell management are studied in parallel. The first one associates place cells with place fields that are given a priori and that are uniformly distributed in the environment. The second one dynamically recruits place cells as exploration proceeds and adjusts the density of such cells to the local complexity of the environment. Both implementations lead to identical results. The article ends with a few predictions about results to be expected in experiments involving simultaneous recordings of multiple cells in the rat hippocampus. PMID- 11007302 TI - Spatial cognition and neuro-mimetic navigation: a model of hippocampal place cell activity. AB - A computational model of hippocampal activity during spatial cognition and navigation tasks is presented. The spatial representation in our model of the rat hippocampus is built on-line during exploration via two processing streams. An allothetic vision-based representation is built by unsupervised Hebbian learning extracting spatio-temporal properties of the environment from visual input. An idiothetic representation is learned based on internal movement-related information provided by path integration. On the level of the hippocampus, allothetic and idiothetic representations are integrated to yield a stable representation of the environment by a population of localized overlapping CA3 CA1 place fields. The hippocampal spatial representation is used as a basis for goal-oriented spatial behavior. We focus on the neural pathway connecting the hippocampus to the nucleus accumbens. Place cells drive a population of locomotor action neurons in the nucleus accumbens. Reward-based learning is applied to map place cell activity into action cell activity. The ensemble action cell activity provides navigational maps to support spatial behavior. We present experimental results obtained with a mobile Khepera robot. PMID- 11007303 TI - Predictions derived from modelling the hippocampal role in navigation. AB - A computational model of the lesion and single unit data from navigation in rats is reviewed. The model uses external (visual) and internal (odometric) information from the environment to drive the firing of simulated hippocampal place cells. Constraints on the functional form of these inputs are drawn from experiments using an environment of modifiable shape. The place cell representation is used to guide navigation via the creation of a representation of goal location via Hebbian modification of synaptic strengths. The model includes consideration of the phase of firing of place cells with respect to the theta rhythm of hippocampal EEG. A series of predictions for behavioural and single-unit data in rats are derived from the input and output representations of the model. PMID- 11007304 TI - The endothelial surface layer. AB - The endothelial lining of blood vessels presents a large surface area for exchange of materials between blood and tissues, and is critically involved in many other processes such as regulation of blood flow, inflammatory responses and blood coagulation. It has long been known that the luminal surface of the endothelium is lined with a glycocalyx, a layer of membrane-bound macromolecules which has been determined by electron microscopy to be several tens of nanometers thick. However, investigations in vivo have indicated the presence of a much thicker endothelial surface layer (ESL), with an estimated thickness ranging from 0.5 microm to over 1 microm, that restricts the flow of plasma and can exclude red blood cells and some macromolecular solutes. The evidence for the existence of the ESL, hypotheses about its composition and biophysical properties, its relevance to physiological processes, and its possible clinical implications are considered in this review. PMID- 11007305 TI - Na+/H+ and CI-/HCO3-antiporters of bovine pigmented ciliary epithelial cells. AB - Medical therapy of glaucoma commonly aims at slowing aqueous humor formation by the ocular ciliary epithelial bilayer, but underlying mechanisms are poorly understood. The first step in secretion is NaCl uptake from the stroma into the pigmented ciliary epithelial (PE) cell layer by electroneutral transporters. After crossing gap junctions into the nonpigmented ciliary epithelial (NPE) cell layer, solute is released into the aqueous humor. Published data have indicated that both paired Na+/H+ and Cl-/HCO3- antiporters and the Na+-K+-2Cl- symporter are involved in net uptake. The molecular identities of the paired antiporters have not been elucidated. We have studied continuously cultured bovine PE cells. Acid-activated 22Na+ uptake was inhibited by cariporide, EIPA (ethyl-isopropyl amiloride) and amiloride, at concentrations characteristic of the NHE-1 isoform. Videomicroscopy of BCECF-loaded PE cells verified the presence of an EIPA inhibitable Na+/H+ antiporter. Removing external Cl- also triggered an alkalinization, which was Na+-independent and could be inhibited by 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Application of hypotonicity followed by return to isotonicity triggered a regulatory volume increase, which was pharmacologically similar to the uptake mechanisms described for intact rabbit ciliary epithelium. Reverse transcriptase polymerase chain reaction (RT PCR) amplification of RNA from the human ciliary body detected expression of the AE2 Cl-/HCO3- exchanger, but not of AE1, cAE3 or bAE3. Immunostaining of bovine PE cells also revealed the presence of AE2 epitope. We conclude that paired NHE-1 Na+/H+ and AE2 Cl-/HCO3- antiporters are important components in the initial step in aqueous humor formation. PMID- 11007306 TI - Structural requirements for determining the substrate affinity of peptide transporters PEPT1 and PEPT2. AB - Peptide transporters PEPT1 and PEPT2 transport numerous compounds including small peptides, peptide-like drugs and nonpeptidic compounds such as valacyclovir. PEPT1 and PEPT2 show low and high affinity for most substrates, respectively, but beta-lactam antibiotics without an alpha-amino group are the only known substrates that prefer PEPT1 to PEPT2. The aim of this study was to compare the recognition and affinity of various substrates between rat PEPT1 and rat PEPT2, and to determine the structural requirements influencing the substrate affinity. [14C]Glycylsarcosine uptake by PEPT1- or PEPT2-expressing transfectant was inhibited by di- and tripeptides, but not by amino acids, tetrapeptides or most cyclic dipeptides. All dipeptides and tripeptides examined showed more potent inhibition of [14C]glycylsarcosine uptake via PEPT2 than via PEPT1, irrespective of their charge and structure. Modification of the alpha-amino group of dipeptides reduced their substrate affinity to both transporters, as compared to unmodified dipeptides, but these dipeptides still showed potent inhibitory effects on PEPT2. Among the nonpeptidic substrates tested, only the eight-amino octanoic acid displayed stronger inhibition of [14C]glycylsarcosine uptake in PEPT1 than in PEPT2. These findings suggest that alpha- or beta-amino carbonyl function is the key structure responsible for the higher affinity for PEPT2 than for PEPT1. PMID- 11007307 TI - Differential calcium responses to the pituitary adenylate cyclase-activating polypeptide (PACAP) in the five main cell types of rat anterior pituitary. AB - We have compared the effects of pituitary adenylate cyclase-activating polypeptide (PACAP-27) on the five main cell types of rat anterior pituitary in primary culture by monitoring changes in cytosolic Ca2+ concentration ([Ca2+]i) in single fura-2-loaded cells. Cells were typed by multiple sequential primary immunocytochemistry at the end of the Ca2+ measurements. PACAP-27 increased [Ca2+]i by three different mechanisms, each one dominant in a given cell type. These involved Ca2+ entry or release from the stores and mediation through different second messenger pathways: (1) stimulation of Ca2+ entry mediated by cAMP was the main mechanism in somatotrophs; (2) Ca2+ release from the intracellular Ca2+ stores mediated by phospholipase C (PLC) was the dominant modality in gonadotrophs; (3) stimulation of Ca2+ entry not mediated by cAMP was the main mechanism in lactotrophs. A minor fraction of somatotrophs (11%) may also use mechanism 3. Corticotrophs and thyrotrophs exhibited weak responses to PACAP (<10% of the cells responded), which in all cases were mediated by mechanism 1. Mechanism 3 represents a novel effect of PACAP which cannot be explained by interaction with the conventional PACAP receptor families. PMID- 11007308 TI - The properties of the Kir6.1-6.2 tandem channel co-expressed with SUR2A. AB - Functional ATP-sensitive K (KATP) channels have an octameric subunit structure with four pore-forming subunits (Kir6.x) and four sulfonylurea receptors (SURx). In the present study, the properties of the heteromeric KATP channel whose pore subunits are composed of Kir6.1 and Kir6.2 were examined using a heterologous expression system. In COS7 cells co-transfected with Kir6.1, Kir6.2 and SUR2A at a ratio of 1:1:2, KATP channels showed various unitary conductances between those of Kir6.1/SUR2A (33.6+/-4.2 pS) and Kir6.2/ SUR2A (67.1+/-1.6 pS). Kir6.1-6.2 tandem protein, constructed by fusing the C-terminus of Kir6.1 to the N-terminus of Kir6.2 with a ten glutamine linker sequence, also formed a channel with an intermediate conductance (58.9+/-1.5 pS). Kir6.2 and Kir6.1-6.2 showed similar sensitivity to ATP4-: half-maximal inhibition (IC50) was obtained at 14.1+/-12.8 microM and 17.6+/-9.6 microM, respectively. In the presence of Mg2+, Kir6. 1-6.2 was significantly less sensitive than Kir6.2 to MgATP (IC50=95.5+/-49.6 microM versus 18.9+/-5.0 microM). These results suggest that Kir6.1 and Kir6.2 are endowed with the potential to form a heteromeric KATP channel, which has a low sensitivity to MgATP. PMID- 11007309 TI - Orexin A activates leptin-responsive neurons in the arcuate nucleus. AB - Orexins, also named hypocretins, are newly described neuropeptides, which are produced almost exclusively in neurons of the lateral hypothalamus and have been shown to increase food intake after intracerebroventricular injection. Leptin, the ob-gene product released from white adipocytes, is suspected to reduce food intake mainly by acting on neurons in the arcuate nucleus of the hypothalamus. Application of orexin A activated 85% (66 out of 78) of all neurons of the rat arcuate nucleus investigated electrophysiologically in an in vitro slice preparation, by a direct excitatory postsynaptic effect. Leptin inhibited electrical activity in 10 out of 22 orexin-sensitive neurons in this brain region and excited only 3 neurons. These data give the first indication as to where and how orexin might interact with the leptin-responsive hypothalamic network. PMID- 11007310 TI - Modulation of fetal and adult acetylcholine receptors by Ca2+ and Mg2+ at developing mouse end-plates. AB - It has long been known that extracellular Ca2+ and Mg2+ modulate synaptic transmission at the neuromuscular junction, acting both pre- and post synaptically. Relevant questions concerning the modulation of acetylcholine (ACh) receptors (AChRs) are however still open: are the fetal (gamma-AChR) and adult (epsilon-AChR) receptors modulated differently? Does the ACh concentration influence the effect of divalent cations? Is the effect on channel open duration dependent on type and concentration of divalent cation? These questions were addressed by studying the modulation of the single-channel behaviour of gamma- and epsilon-AChRs by Ca2+ and Mg2+ at the endplate of muscle fibres acutely dissociated from 12- to 14-day-old mice. Ca2+ reduced the conductances of the two receptor channels comparably. Mg2+ had a stronger effect than Ca2+ and reduced the conductance of epsilon-AChR significantly more than that of gamma-AChR. With 0.1 microM ACh, Ca2+ and Mg2+ increased the mean open duration of gamma- and epsilon-AChR channels comparably. At 100 microM ACh, gamma- and epsilon-AChR channels opened in bursts of strikingly similar duration, which was unaffected by divalent cations. These findings indicate that Ca2+, and even more so Mg2+, may regulate synaptic transmission by modulating the function of AChRs in addition to the well-established effects on transmitter release. PMID- 11007311 TI - Nitric oxide uncouples gap junctions of supporting Deiters cells from Corti's organ. AB - Supporting cells of Corti's organ are electrically coupled via gap junctions. They probably serve to maintain the unique cochlear environment that is required for normal sensory function. In this study we used input capacitance measurements under whole-cell voltage-clamp conditions to evaluate the effects of nitric oxide on gap junctional communication between pairs of isolated supporting Deiters cells. We show that the nitric oxide (NO) donor sodium nitroprusside causes the uncoupling of Deiters cells, and that an NO synthase inhibitor blocks the effect. The cGMP analogue 8-bromo-cGMP also uncouples Deiters cells. With either treatment, the input capacitance of pairs of Deiters cells drops to single-cell levels within minutes of application, indicative of electrical uncoupling. We surmise that the NO/cGMP pathway may serve to modulate normal cochlear homeostasis and possibly plays a role in ototoxic mechanisms. PMID- 11007312 TI - Intracellular pH regulatory mechanism in a human renal proximal cell line (HKC 8): evidence for Na+/H+ exchanger, CI-/HCO3- exchanger and Na+-HCO3- cotransporter. AB - In the present study we investigated whether an immortalized human renal proximal cell line, HKC-8, expresses a recently cloned Na+-HCO3- cotransporter (NBC-1) and, if so, which isoform (kNBC-1 from kidney or pNBC-1 from pancreas) is expressed in this cell line. Cell pH (pHi) measurements using a pH-sensitive fluorescence probe in the absence of HCO3-/CO2 revealed the presence of a Na+/H+ exchanger that required high concentrations of amiloride for full inhibition. In the presence of HCO3-/CO2 another pHi recovery process, dependent on Na+ but independent of Cl-, was identified. This process was electrogenic and was inhibited by 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), being consistent with the Na+-HCO3- cotransporter. In addition, the pHi responses to Cl- removal were compatible with the presence of a Na+-independent Cl-/HCO3- exchanger that was also inhibited by DIDS. Reverse transcriptase polymerase chain reaction (RT-PCR) using primers designed for specific and common regions detected mRNAs of both kNBC-1 and pNBC-1 and Western blot analysis confirmed the expression of NBC-1 protein. These results indicate that HKC-8 has transport activities similar to intact proximal tubules and also suggest that both kNBC-1 and pNBC-1 may contribute to the Na+-HCO3- cotransport activity in this cell line. PMID- 11007313 TI - Effect of glycolysis on pancreatic microcirculation and cellular functions in anesthetized rats. AB - It has been reported that aerobic glycolysis in the pancreas contributes less than 10% to oxidative phosphorylation based on in vitro experiments using pancreatic tissue segments. However, its contribution to aerobic glycolysis in vivo remains uncertain. We investigated the effect of inhibiting glycolysis on O2 metabolism in microvessels, exocrine enzyme secretion, and the blood glucose level in the pancreas of anesthetized rats in vivo. Inhibition of glycolysis, by superfusing the pancreas of anesthetized rats with 2-deoxyglucose (10 mM) or sodium fluoride (2 mM), significantly decreased O2 release from erythrocytes flowing in the microvessels by 30-40%. Inhibiting glycolysis did not affect the exocrine secretion of pancreatic juice but decreased the secretion of total protein by approximately or = to 40%. Inhibiting glycolysis decreased blood glucose levels by approximately or = to 40% and increased glucagon release twofold. Aerobic glycolysis may play more important roles in the regulation of O2, metabolism, pancreatic exocrine enzyme secretion and the blood glucose level in rat pancreas. PMID- 11007314 TI - Hypoxia and fatigue-induced modification of function and proteins in intact and skinned murine diaphragm muscle. AB - Fatigue studies of isolated, intact muscles typically utilize solutions saturated with O2. However, under in vivo fatiguing conditions, less oxygen is delivered to the muscles and they actually experience hypoxia. No studies to date have correlated the effects of acute hypoxia on the isometric contractile properties of intact muscles, skinned fibers isolated from the same muscles, and the cellular content of specific muscle proteins. Therefore, we have studied the effects of in vitro acute hypoxia on the fatigability of intact diaphragm muscle strips and on the isometric contractile properties of single Triton-skinned fibers isolated from control and hypoxic diaphragm muscles. We found that hypoxia and fatiguing stimulation per se affect the tetanic force of intact muscle strips without exhibiting any significant deleterious effects on the calcium-activated force of skinned muscle fibers dissected from the intact muscles. In contrast, fatiguing stimulation under hypoxic conditions decreased both the tetanic force of muscle strips and the calcium-activated force of skinned muscle fibers. Gel electrophoresis of muscles subjected to hypoxia and hypoxic-fatigue revealed that there is a significant reduction in three protein bands when compared to control muscles. Protein modification may be the underlying mechanism of muscle fatigue under physiologic conditions. PMID- 11007315 TI - Na/K pump alpha subunit expression in rabbit ventricle and regional variations of intracellular sodium regulation. AB - We examined the isoform distribution and expression of the alpha subunit of the Na/K-ATPase in the left ventricular muscle of rabbit heart in order to determine whether previously reported regional differences in intracellular sodium regulation derive from differences in pump expression. Immunohistochemical techniques show that only the alpha1 isoform is present in rabbit ventricle; therefore, regional variation in isoform distribution is not possible. Western blots of samples taken from subendocardial and sub-epicardial regions confirm the absence of alpha2 and alpha3 isoforms but also show that levels of the alpha1 isoform do not differ significantly in the two regions. The ratio of densitometric readings from blot bands was 1.18+/-0.17 (epicardial:endocardial; mean +/-SEM). Measurements of fully activated pump current in voltage-clamped cells were achieved by dialysing the cell via the patch pipette with 50 mM Na and applying 1 mM dihydroouabain. The current measured was 0.16+/-0.02 pA/pF in epicardial cells and 0.17+/-0.02 pA/pF in endocardial cells. These results indicate that the capacity of cells from the two regions to generate sodium efflux is identical. Regional differences in intracellular sodium regulation, therefore, are more likely to arise from differences in influx of sodium. PMID- 11007316 TI - Cs+ block of the cardiac muscarinic K+ channel, GIRK1/GIRK4, is not dependent on the aspartate residue at position 173. AB - Cs+ block of GIRK1/GIRK4 expressed in Xenopus oocytes has been investigated. It has been reported that a negatively charged aspartate residue at position 172 in IRK1 is responsible for Cs+ block of the channel. IRK1, a homotetramer, has four aspartate residues at this position. GIRK1/GIRK4 is a heterotetramer and has two aspartate residues at the equivalent position (GIRK1-D173) and, consequently, it should be less sensitive to Cs+. Cs+ caused voltage-dependent block of GIRK1/GIRK4 current (measured with the two-microelectrode voltage-clamp technique). The apparent fraction of the electrical field through which Cs+ moves in order to reach its site of block (delta approximately equals 1.66) is comparable to that in IRK1, suggesting that Cs+ binds to a similar site in the two channels. GIRK1/GIRK4 was less sensitive than IRK1 to Cs+ -the Kd was 3.0-8.5 times greater and at potentials more negative than approximately or = to 130 mV there was voltage-dependent relief of block of GIRK1/GIRK4 (not the case with IRK1). However, the mutations GIRK1-D173A and GIRK1-D173Q increased the sensitivity of the channel to Cs+, while adding a negatively charged aspartate residue to GIRK4 at the equivalent position (GIRK4-N 79D) decreased Cs+ sensitivity. GIRK1-D173 cannot be the site of Cs+ block of GIRK1/GIRK4. PMID- 11007317 TI - Changes in the Ca2+-activation characteristics of demembranated rat single muscle fibres after prolonged incubation at room temperature in the presence and absence of DTT and protease inhibitors. AB - Prolonged incubation (24 h) of chemically skinned rat muscle preparations in rigor solutions at room temperature and in the absence of reducing agents and protease inhibitors modified the Ca2+-activation characteristics of the contractile machinery. In the absence of reducing agents and protease inhibitors, the contraction threshold for incubated fibres was shifted to lower Ca2+ concentrations and the steepness of the steady-state force-pCa (-log10)[Ca2+]) curve was decreased compared to that of control muscle fibres. Mean myosin ATPase activity under these conditions was significantly lowered by a factor of 2.7. Fibres incubated in the presence of 10 mM dithiothreitol (DTT) and protease inhibitors (100 microM pepstatin A/200 microM leupeptin) produced a maximum Ca2+ activated force per cross-sectional area that compared favourably with that of freshly dissected muscle fibres and there were no changes in the other contractile activation characteristics. Intermediate responses were obtained when fibres were incubated in the presence of either DTT or protease inhibitors. MgATPase activities of incubated preparations increased significantly following the addition of protease inhibitors and/or DTT to the incubation medium. Taken together, these results suggest that in the presence of DTT and protease inhibitors, most contractile properties are maintained at levels seen in fresh mechanically skinned fibres. The extended viability of this preparation and its closely related properties with fresh muscle fibres make it a useful model for experiments requiring longer term incubations with biological agents. PMID- 11007318 TI - Volume-sensitive KCI cotransport associated with human cervical carcinogenesis. AB - This study investigates the volume-sensitive KCI cotransporter (KCC) in various types of human cervical epithelial cell, testing the hypothesis that cervical malignancy is accompanied by differential expression of volume-sensitive KCC. Normal human cervical epithelial cells have KCCs which are quiescent in normal physiological conditions and are relatively refractory to hypotonic stress. By contrast, cervical cancer cells have KCCs which are also nearly quiescent in normal physiological conditions but high transport rates are observed in response to hypotonic challenge. Using isoform-specific primers, mRNA transcripts of KCC1, KCC3 and KCC4 were identified by reverse transcriptase polymerase chain reaction (RT-PCR) in several types of cervical cell, and confirmed by digestion with specific restriction endonucleases. By semiquantitative RT-PCR with beta-actin as the internal standard, the results indicate that cervical carcinogenesis is accompanied by the up-regulation of mRNA transcripts in KCC1, KCC3 and KCC4. [(Dihydroindenyl)oxy] alkanoic acid (DIOA), a KCC inhibitor, blocked both the regulatory volume decrease (RVD) process and volume-sensitive 86Rb+ efflux from cervical cancer cells in a dose-dependent manner. The volume-sensitive 86Rb+ efflux from cervical cancer cells was also blocked by two protein phosphatase inhibitors, calyculin A and okadaic acid, with IC50 values of 0.8 and 6 nM, respectively. Conversely, protein kinase inhibitors, chelerythrine and staurosporine, increased Cl- dependent 86Rb+ efflux. NEM (1 mM) led to a fivefold stimulation of 86Rb+ efflux which was totally Cl- dependent in cervical cancer cells. Hypotonicity could not stimulate any further 86Rb+ efflux after NEM treatment. These results indicate that the volume-sensitive KCC in cervical cancer cells plays a significant role in volume regulation and that the activities are modulated by a phosphorylation cascade. Taken together with our previous studies, we suggest the volume-regulatory ion channels and the co transport systems work synergistically for volume regulation in human cervical cancer cells. PMID- 11007319 TI - Ornithine metabolism along the female mouse nephron: localization of ornithine decarboxylase and ornithine aminotransferase. AB - The fate of ornithine in the nephron of the female OF-1 Swiss mouse remains unknown. The aim of the present study was to identify the nephron segments containing the key enzymes involved in ornithine metabolism: ornithine decarboxylase (ODC) and ornithine aminotransferase (OAT). Viable tubules isolated by microdissection were incubated with [1-14C]ornithine to study the oxidative pathway. Other tubules were permeabilized to measure the ODC activity. Ornithine was decarboxylated in all intact tubules. Gabaculine, a suicide inhibitor of OAT, and rotenone sharply decreased the production of 14CO2 from [1-14C]ornithine. No ODC activity was found in permeabilized tubules isolated from untreated mice. Testosterone increased ODC activity in the proximal tubule substantially and to a minor extent in other nephron segments. In situ hybridization showed ODC messenger ribonucleic acid (mRNA) to be absent in kidneys of untreated females but abundant in the cortex and the outer stripe of the outer medulla of testosterone-treated female mice. The whole proximal tubule contained a great density of silver grains corresponding to ODC mRNA. In conclusion, no basal ODC activity was found in the nephron of female mice. The testosterone-inducible ODC is localized mainly in the proximal tubule, but is also present in distal tubules and collecting ducts. OAT is distributed along the whole nephron, but its activity is higher in proximal tubules than in distal tubules. PMID- 11007320 TI - Potassium efflux triggered by P2Y purinoceptor activation in cultured pituicytes. AB - We have previously investigated the effects of extracellular ATP on the concentration of free cytosolic calcium ([Ca2+]i) from rat cultured neurohypophysial astrocytes (pituicytes). We demonstrated that ATP acts via a P2Y receptor to increase [Ca2+]i. In the present study, we examine the effect of ATP on K+ efflux using 86Rb+ as an isotopic tracer, in order to characterize the possible presence of a Ca2+-activated K+ conductance and to establish the implications of pituicytes in the regulation of stimulus-secretion coupling. ATP evoked an increase in 86Rb+ efflux from cultured pituicytes. This effect was Ca2+ dependent, as indicated by the unresponsiveness of cells loaded with BAPTA/AM (20 microM). Furthermore, the effect of ATP was mimicked by 2-methylthio-adenosine-5' triphosphate (2MeSATP), a P2 purinoceptor agonist, and abolished by Reactive Blue 2 (RB-2), a selective P2Y antagonist, implying a role for the P2Y purinoreceptor. A pharmacological study revealed that Ba2+ and tetraethylammonium (TEA), two inhibitors of K+ channels, both strongly reduced the ATP-stimulated 86Rb+ efflux. In addition, the effect of ATP was modulated by different peptidic toxins. Apamin (100 nM), an inhibitor of the small-conductance Ca2+-activated K+ channels, partly blocked ATP-induced 86Rb+ efflux. Leiurus quinquestriatus hebraeus (LQH) scorpion venom (20 microg/ml) and Buthus tamulus (BT) scorpion venom (20-200 microg/ml) inhibited ATP-induced 86Rb+ efflux. The specificity of the effects of the crude venoms was checked using charybdotoxin (100 nM) and iberiotoxin (1 pM), which are the active toxins extracted from the LQH and BT venoms, respectively. These data indicate the involvement of several types of Ca2+-activated K+ channels in the ATP-dependent K+ efflux, and lead to the proposal that, in the neurohypophysis, extracellular ATP released by nerve terminals may act directly on the pituicytes and induce a K+ efflux via a P2Y purinoreceptor. PMID- 11007321 TI - Determinants of the difference between expired and core temperatures: effect of a breath-hold. AB - After a 30-s breath-hold (BH), expired temperature (TE) does not reach core temperature. One explanation is that the gas in the airways is not in thermal equilibrium with the airway walls. This possibility was eliminated by comparing TE in six subjects breathing either helium-oxygen or air after a BH. Another possibility is that the airway walls and surrounding tissues have sufficient thermal inertia to slow down thermal equilibrium during BH. This was checked by measuring oral and upper esophageal temperatures after cooling or heating the airways. It took more than 2 min for these temperatures to recover their steady state value. Six subjects were requested to perform a long apnea after hyperventilating for 1 min and then taking a single breath of 100% oxygen. TE was still lower than core temperature after a 1-min BH, and there was no difference after a 2-min BH. The difference between expired and core temperatures during BH thus appears to be due to the thermal inertia of the airways and their surrounding tissues. PMID- 11007322 TI - Modulation of a cloned human A-type voltage-gated potassium channel (hKv1.4) by the protein tyrosine kinase inhibitor genistein. AB - A cloned, human, A-type, voltage-gated potassium channel (hKv1.4) was expressed transiently in Chinese hamster ovary cells and the effects of the broad-spectrum tyrosine kinase inhibitor genistein on hKv1.4 were studied using the whole-cell patch-clamp recording method. Genistein (up to 50 microM) reversibly reduced the peak currents of hKv1.4 by 44.9+/-12%. In addition, genistein markedly slowed the activation kinetics (time constant tau(a)) of hKv1.4. At +50 mV, tau(a) increased from 1.8+/-0.3 to 5.0+/-0.6 ms (P<0.01). The effect of genistein on the channel inactivation kinetics (time constant tau(i)) was more complex, in that tau(i) was increased significantly at lower step potentials but unaltered at +50 mV or more depolarized potentials. Tail current analysis showed that genistein had no effect on the kinetics of deactivation (time constant tau(d)), but shifted the steady state activation curve significantly to the right by about 15 mV (potential for half-maximal activation, V1/2, changed from -7.4+/-4.4 to +7.7+/-2.7 mV) with a moderate change in the slope (k) of the curve (from 17.4+/-2.2 to 23+/-1.0 mV, P<0.05). Genistein slightly altered the slope of the steady-state inactivation curve from -5.5+/-0.4 to -7.5+/-0.4 mV (P<0.01). The recovery rate from inactivation was not altered by genistein. The tyrosine phosphatase inhibitor orthovanadate (1 mM) alone had little impact on current amplitude or channel kinetics. However, orthovanadate significantly, but not completely, blocked the effect of genistein on current amplitude (by 25.5%) and kinetics (by 67.1%). Daidzein (up to 50 microM), an inactive analogue of genistein, had no effect on current amplitude or kinetics. In contrast to genistein, another tyrosine kinase inhibitor, herbimycin A, had little effect on the channel peak amplitude or kinetics. In addition, genistein had a similar impact on the channel peak current amplitude and kinetics in cells with or without pre-treatment with herbimycin A (10 microM). The data suggest that genistein-induced inhibition of tyrosine phosphorylation may not be the exclusive mechanism by which hKv1.4 is down regulated and channel gating affected. Genistein may produce a non-catalytic blockade of this channel. PMID- 11007324 TI - A new arthropod from the Silurian Konservat-Lagerstatte of Herefordshire, UK. AB - A small, non-biomineralized, macrophagous arthropod with chelicerate affinities, Offacolus kingi gen. et sp. nov., from the Silurian (Wenlock Series) of Herefordshire, UK, is described. The dorsal exoskeleton comprises an arch-like cephalic shield, a thorax of three free tergites and a triangular posterior tagma of five fused tergites, the last with a stout postero-dorsally directed medial spine. Seven pairs of appendages beneath the cephalic shield surround a postero medially sited oral cavity on the ventral surface of the head. Appendages I and, probably II are uniramous and project antero-ventrally; I was sensory and II sensory and/or ambulatory. Appendages III-VI are biramous, each with an antero ventrally projecting ramus and a robust, highly geniculate, horizontally oriented ramus that projects through an anterior gape. The former rami were ambulatory and the latter have spinose terminal podomeres and functioned as a unit for trapping food and transferring it towards the oral cavity. Appendage VII, which is probably uniramous, is posteroventrally directed and flap like. Each tergite of the thorax and posterior tagma covers at least a pair (probably two pairs) of probably biramous appendages with each ramus flap like and setose. PMID- 11007323 TI - Evolutionary novelties in islands: Drosophila santomea, a new melanogaster sister species from Sao Tome. AB - The finding of new melanogaster sister species may help us in understanding more about how the emergence of genetic novelties, particularly in insular habitats, can result in speciation. Here we report on the discovery of Drosophila santomea, which is the first melanogaster sibling found off West-equatorial Africa, on Sao Tome, one of the Gulf of Guinea islands. Although the eight other melanogaster sister species are remarkably conservative in their morphology except for their terminalia, the new find has a morphological trait distinguishing it from all of these: a pure yellow body coloration of both sexes without the normal black abdominal banding. Evidence from the terminalia, polytene and mitotic chromosomes, period gene and allozymes are provided indicating that it is nonetheless the nearest relative of Drosophila yakuba with which it coexists on the island. The new find is a clear-cut taxon as shown by the production of sterile male hybrids, eventually with developmental defects, in both directions of cross with yakuba and by the existence of an altitudinal divide accompanied by a hybrid zone at mid-elevation on the island. Molecular and karyotypic data further support this conclusion. In contrast to the significant divergence of their nuclear DNAs, an intriguing similarity in their cytochrome b sequences was observed indicating a recent coalescence common to santomea, yakuba and also teissieri cytoplasms. These were shown to harbour the same Wolbachia endosymbiotic bacteria which could possibly be responsible for mitochondrial DNA hitchhiking across the species barrier. PMID- 11007325 TI - Non-random dispersal in the butterfly Maniola jurtina: implications for metapopulation models. AB - The dispersal patterns of animals are important in metapopulation ecology because they affect the dynamics and survival of populations. Theoretical models assume random dispersal but little is known in practice about the dispersal behaviour of individual animals or the strategy by which dispersers locate distant habitat patches. In the present study, we released individual meadow brown butterflies (Maniola jurtina) in a non-habitat and investigated their ability to return to a suitable habitat. The results provided three reasons for supposing that meadow brown butterflies do not seek habitat by means of random flight. First, when released within the range of their normal dispersal distances, the butterflies orientated towards suitable habitat at a higher rate than expected at random. Second, when released at larger distances from their habitat, they used a non random, systematic, search strategy in which they flew in loops around the release point and returned periodically to it. Third, butterflies returned to a familiar habitat patch rather than a non-familiar one when given a choice. If dispersers actively orientate towards or search systematically for distant habitat, this may be problematic for existing metapopulation models, including models of the evolution of dispersal rates in metapopulations. PMID- 11007326 TI - Developmental trade-offs and life histories: strategic allocation of resources in caddis flies. AB - Resource allocation trade-offs during development are potentially very important in the evolution of organism morphology and life-history strategy However, they have rarely been demonstrated empirically. To what extent the division of limited resources between growing organs is a consequence of particular developmental pathways or varies strategically in line with life-history predictions is unknown. It has been demonstrated in a number of holometabolous insects that altering the resources available at pupation changes the pattern of allocation to adult tissues, but this has not been examined in a life-history context. Using caddis flies (Trichoptera), we show here that the effect of depleted larval resources on the pattern of somatic and reproductive investment is not fixed but varies between species with different life-history patterns. In particular, we demonstrate that, in a long-lived species, thorax size is preserved, which contrasts with the pattern previously observed in a short-lived species. That the adult body can be differentially altered by the same resource depletion in the larvae demonstrates that the allocation of resources amongst body parts is not a consequence of fixed pathways during development. Rather, the allocation of resources during development can occur in a manner consistent with the minimization of the effects on adult fitness. PMID- 11007327 TI - Lifetime success and interactions of farm salmon invading a native population. AB - Farm Atlantic salmon escape and invade rivers throughout the North Atlantic annually, which has generated growing concern about their impacts on native salmon populations. A large-scale experiment was therefore undertaken in order to quantify the lifetime success and interactions of farm salmon invading a Norwegian river. Sexually mature farm and native salmon were genetically screened, radio tagged and released into the River Imsa where no other salmon had been allowed to ascend. The farm fishes were competitively and reproductively inferior, achieving less than one-third the breeding success of the native fishes. Moreover, this inferiority was sex biased, being more pronounced in farm males than females, resulting in the principal route of gene flow involving native males mating with farm females. There were also indications of selection against farm genotypes during early survival but not thereafter. However, evidence of resource competition and competitive displacement existed as the productivity of the native population was depressed by more than 30%. Ultimately, the lifetime reproductive success (adult to adult) of the farm fishes was 16% that of the native salmon. Our results indicate that such annual invasions have the potential for impacting on population productivity, disrupting local adaptations and reducing the genetic diversity of wild salmon populations. PMID- 11007328 TI - Differential effects of endoparasitism on the expression of carotenoid- and melanin-based ornamental coloration. AB - The striking diversity of sexual dimorphisms in nature begs the question: Why are there so many signal types? One possibility is that ornamental traits convey different sets of information about the quality of the sender to the receiver. The colourful, pigmented feathers of male birds seem to meet the predictions of this hypothesis. Evidence suggests that carotenoid pigmentation reflects the nutritional condition of males during moult, whereas in many instances melanin pigmentation is a reliable indicator of social status. However, as of yet there have been no experimental tests to determine how these two ornament types respond to the same form of environmental stress. In this study, we tested the effect of endoparasitic infection by intestinal coccidians (Isospora sp.) on the expression of both carotenoid- and melanin-based ornamental coloration in captive male American goldfinches (Carduelis tristis). We found that the carotenoid-based plumage and bill coloration of parasitized males was less saturated than that developed by unparasitized males, but that the brightness and size of melanin based black caps did not differ between the groups. These findings provide the most robust empirical support to date for the notion that carotenoid and melanin ornaments reveal different information to conspecifics. PMID- 11007329 TI - Androgen levels and female social dominance in Lemur catta. AB - Morphological and behavioural traits which improve agonistic power are subject to intrasexual selection and, at the proximate level, are influenced by circulating androgens. Because intrasexual selection in mammals is more intense among males, they typically dominate females. Female social dominance is therefore unexpected and, indeed, rare. Ring-tailed lemurs (Lemur catta) are sexually monomorphic primates in which all adult females dominate all males. The goal of our study was to test the prediction that female dominance in this species is associated with high androgen levels. Using two captive groups, we collected data on agonistic behaviour and non-invasively assessed their androgen concentrations in faeces and saliva by enzyme immunoassay. We found that adult female L. catta do not have higher androgen levels than males. However, during the mating season there was a twofold increase in both the androgen levels and conflict rates among females. This seasonal increase in their androgen levels was probably not due to a general increase in ovarian hormone production because those females showing the strongest signs of follicular development tended to have low androgen concentrations. At the individual level neither the individual aggression rates nor the proportion of same-sexed individuals dominated were correlated with their androgen levels. We conclude that female dominance in ring-tailed lemurs is neither based on physical superiority nor on high androgen levels and that it is equally important to study male subordination and prenatal brain priming effects for a complete understanding of this phenomenon. PMID- 11007330 TI - Strategic allocation of ejaculates by male Adelie penguins. AB - Sperm competition theory suggests that males should strategically allocate sperm to those females that will bring them the best possible genetic returns. Although males of a number of species of insects and fishes have been shown to allocate sperm strategically, we provide, to our knowledge, the first evidence that an avian species is also capable of allocating ejaculates. Male Adelie penguins (Pygoscelis adeliae) are more likely to transfer sperm during extra-pair copulations (EPCs) than during pair copulations. We investigated the question of how males allocate ejaculates within the constraints of limited sperm availability and found (i) that males that engaged in EPC attempts ejaculated less often when copulating with their social partner than males that made no EPC attempts, and (ii) that there was no difference between males that were involved in failed EPC attempts and those that were involved in successful EPCs in the proportion of copulations that resulted in sperm transfer. These results indicate that males achieve strategic allocation of sperm within the constraints of limited sperm availability by withholding ejaculates from their social partners. PMID- 11007331 TI - Sperm competition games played by dimorphic male beetles: fertilization gains with equal mating access. AB - Alternative mating tactics can generate asymmetry in the sperm competition risk between males within species. Theory predicts that adaptations to sperm competition should arise in males facing the greater risk. This prediction is met in the dung beetle Onthophagus binodis where minor males which sneak copulations have a greater expenditure on the ejaculate. In its congener Onthophagus taurus there is a reduced asymmetry in sperm competition risk such that both tactics have equal ejaculate expenditure. We used the irradiated male technique to test whether adaptations to sperm competition in minor males result in higher paternity. We found that for both species, on average, each of two males gained equal numbers of fertilizations, confirming the assumption that sperm compete in a raffle. There were no differences in the sperm competition success of major and minor males in O. taurus as predicted from their equal expenditure on their ejaculate. Contrary to expectations, there were also no differences in fertilization success between the male tactics in O. binodis. Thus, in O. binodis minor males must expend more on their ejaculate in order to obtain the same fertilization gains as major males. PMID- 11007332 TI - Sexually transmitted diseases in polygynous mating systems: prevalence and impact on reproductive success. AB - Studies of disease in relation to animal mating systems have focused on sexual selection and the evolution of sexual reproduction. Relatively little work has examined other aspects of ecological and evolutionary relationships between host social and sexual behaviour, and dynamics and prevalence of infectious diseases; this is particularly evident with respect to sexually transmitted diseases (STDs). Here, we use a simulation approach to investigate rates of STD spread in host mating systems ranging from permanent monogamy to serial polygyny or polyandry and complete promiscuity. The model assumes that one sex (female) is differentially attracted to the other, such that groups of varying size are formed within which mating and disease transmission occur. The results show that equilibrium disease levels are generally higher in females than males and are a function of variance in male mating success and the likelihood of a female switching groups between mating seasons. Moreover, initial rates of disease spread (determining whether an STD establishes in a population) depend on patterns of host movement between groups, variance in male mating success and host life history (e.g. mortality rates). Male reproductive success can be reduced substantially by a sterilizing STD and this reduction is greater in males that are more 'attractive' to females. In contrast, females that associate with more attractive males have lower absolute fitness than females associating with less attractive males. Thus, the potential for STDs to act as a constraint on directional selection processes leading to polygyny (or polyandry) is likely to depend on the details of mate choice and group dynamics. PMID- 11007333 TI - Egg maturation, egg resorption and the costliness of transient egg limitation in insects. AB - Although there is widespread agreement that the cost of oviposition underlies selective oviposition in insects, there is no consensus regarding which factors mediate the cost of oviposition. Models have suggested that egg costs are often paramount in those insects that do not continue to mature eggs during the adult stage (pro-ovigenic insects). Here we address the hypothesis that egg costs are generally less significant in synovigenic insects, which can replenish oocyte supplies through continuous egg maturation. A dynamic optimization model based on the biology of a highly synovigenic parasitoid, Aphytis aonidiae, suggests that the maximum rate of egg maturation is insufficient to balance the depletion of eggs when opportunities to oviposit are abundant. Transient egg limitation therefore occurs, which imposes opportunity costs on reproducing females. Thus, whereas the most fundamental constraint acting on the lifetime reproductive success of pro-ovigenic species is the fixed total number of eggs that they carry at eclosion, the most fundamental constraint acting on a synovigenic species is the maximum rate of oocyte maturation. Furthermore, the ability of synovigenic species to reverse the flow of nutrients from the soma to oocytes (i.e. egg resorption) has a dramatic influence on the cost of oviposition. Whereas females in hostrich environments may experience oviposition-mediated egg limitation, females in host-poor environments may experience oosorption-mediated egg limitation. Both forms of egg limitation are costly. Contrary to initial expectations, the flexibility of resource allocation that typifies synovigenic reproduction actually appears to broaden the range of conditions under which costly egg limitation occurs. Egg costs appear to be fundamental in mediating the trade-off between current and future reproduction, and therefore are an important factor favouring selective insect oviposition. PMID- 11007334 TI - A modelling approach to vaccination and contraception programmes for rabies control in fox populations. AB - In a previous study, three of the authors designed a one-dimensional model to simulate the propagation of rabies within a growing fox population; the influence of various parameters on the epidemic model was studied, including oral vaccination programmes. In this work, a two-dimensional model of a fox population having either an exponential or a logistic growth pattern was considered. Using numerical simulations, the efficiencies of two prophylactic methods (fox contraception and vaccination against rabies) were assessed, used either separately or jointly. It was concluded that far lower rates of administration are necessary to eradicate rabies, and that the undesirable side-effects of each programme disappear, when both are used together. PMID- 11007335 TI - Host specificity in avian blood parasites: a study of Plasmodium and Haemoproteus mitochondrial DNA amplified from birds. AB - A fragment of the mitochondrial cytochrome b gene of avian malaria (genera Haemoproteus and Plasmodium) was amplified from blood samples of 12 species of passerine birds from the genera Acrocephalus, Phylloscopus and Parus. By sequencing 478 nucleotides of the obtained fragments, we found 17 different mitochondrial haplotypes of Haemoproteus or Plasmodium among the 12 bird species investigated. Only one out of the 17 haplotypes was found in more than one host species, this exception being a haplotype detected in both blue tits (Parus caeruleus) and great tits (Parus major). The phylogenetic tree which was constructed grouped the sequences into two clades, most probably representing Haemoproteus and Plasmodium, respectively. We found two to four different parasite mitochondrial DNA (mtDNA) haplotypes in four bird species. The phylogenetic tree obtained from the mtDNA of the parasites matched the phylogenetic tree of the bird hosts poorly. For example, the two tit species and the willow warbler (Phylloscopus trochilus) carried parasites differing by only 0.6% sequence divergence, suggesting that Haemoproteus shift both between species within the same genus and also between species in different families. Hence, host shifts seem to have occurred repeatedly in this parasite host system. We discuss this in terms of the possible evolutionary consequences for these bird species. PMID- 11007336 TI - Fatal attraction in rats infected with Toxoplasma gondii. AB - We tested the hypothesis that the parasite Toxoplasma gondii manipulates the behaviour of its intermediate rat host in order to increase its chance of being predated by cats, its feline definitive host, thereby ensuring the completion of its life cycle. Here we report that, although rats have evolved anti-predator avoidance of areas with signs of cat presence, T. gondii's manipulation appears to alter the rat's perception of cat predation risk, in some cases turning their innate aversion into an imprudent attraction. The selectivity of such behavioural changes suggests that this ubiquitous parasite subtly alters the brain of its intermediate host to enhance predation rate whilst leaving other behavioural categories and general health intact. This is in contrast to the gross impediments frequently characteristic of many other host parasite systems. We discuss our results in terms of their potential implications both for the epidemiology of toxoplasmosis and the neurological basis of anxiety and cognitive processes in humans and other mammals. PMID- 11007337 TI - Solvent related chronic encephalopathy. PMID- 11007338 TI - Criteria for solvent-induced chronic toxic encephalopathy: a systematic review. AB - In 1985, a WHO Working Group presented diagnostic criteria and a classification for solvent-induced chronic toxic encephalopathy (CTE). In the same year, the "Workshop on neurobehavioral effects of solvents" in Raleigh, N.C., USA introduced a somewhat different classification for CTE. The objective of this review is to study the diagnostic procedures that are used to establish the diagnosis of CTE, and the extent to which the diagnostic criteria and classification of the WHO, and the classification of the Raleigh Working Group, are applied. A systematic search of studies on CTE was performed, and the diagnostic criteria and use of the WHO and Raleigh classifications were listed. We retrieved 30 original articles published in English from 1985 to 1998, in which CTE was diagnosed. Only two articles did not report the duration of solvent exposure. The type of solvent(s) involved was described in detail in four articles, poorly in 17 articles, and not at all in nine articles. Tests of general intelligence were used in 19 articles, and tests of both attention and mental flexibility and of learning and memory were used in 18 articles. Exclusion, by interview, of potentially confounding conditions, such as somatic diseases with central nervous effects and psychiatric diseases, was reported in 21 and 16 articles, respectively. In only six of the articles were both the WHO diagnostic criteria and the WHO or Raleigh classifications used. In the future, parameters of exposure, psychological test results, and use of medication that possibly affects psychological test results should always be described. We list some advantages and disadvantages of the Raleigh and WHO classifications. To aid inter-study comparisons, the diagnosis of CTE should be categorized and reported according to an internationally accepted classification. PMID- 11007339 TI - Exposure and acute exposure-effects before and after modification of a contaminated humidification system in a synthetic-fibre plant. AB - OBJECTIVE: Follow-up study of exposure and acute exposure-effects after modification to steam humidification of a contaminated cold water system which had caused an outbreak of humidifier fever in a synthetic-fibre plant. METHODS: Before and after modification of the system aerobiological measurements were performed. Concentrations of fungi and bacteria, in colony forming units (cfu) per m3, were measured by stationary air sampling with an Andersen sampler. Endotoxin levels (pg/m3) were determined by Limulus Amoebocyte Lysate (LAL) assay in pooled dust from personal air sampling. An indication of exposure levels of oil-mist was obtained by monitoring with a direct reading optical photometer. Changes as acute exposure-effects in spirometry and white blood cell count, during an afternoon shift were compared in exposed and non-exposed workers before and after modification. RESULTS: Measured levels of fungi, total bacteria, Gram negative bacteria and endotoxins both before and after modification were below levels which would be expected to be associated with the exposure-effects. However, after modification, we found that the statistically significant differences in levels of bacteria and endotoxins with a department without humidification no longer existed. Mean oil-mist concentrations were below 1 mg/m3, with short-time peak exposure during certain tasks of up to 5 mg/m3. Before modification, in exposed workers there was significantly more decline of spirometry, and more increase of white blood cell count during the first afternoon shift, compared with non-exposed workers. In exposed workers, the white blood cell count increase was positively associated with decline of spirometry. After modification, differences between exposed and non-exposed workers no longer existed. CONCLUSION: Follow-up investigation of acute exposure-effects demonstrated the effectiveness of remedial actions taken against a contaminated humidification system. Follow-up of exposure-effects in particular is recommended when there is doubt about the interpretation of exposure measurements. PMID- 11007340 TI - Follow-up study of workers in a nylon carpet yarn plant after remedial actions taken against a contaminated humidification system. AB - OBJECTIVE: To investigate the effectiveness of remedial actions taken against a contaminated humidification system, after an outbreak of humidifier disease in a nylon carpet yarn plant. METHODS: Two and 6 years after modification, a follow-up investigation of a stratified (age, smoking habits) sample of exposed (n = 75) and non-exposed (n = 56) workers was carried out. Outcome-parameters were: respiratory symptoms, spirometry, skin tests and serology with various humidifier fungi. In addition, the yearly incidence of work-disability because of chronic respiratory disease during the 2 years before, and 11 years after the modification was compared. RESULTS: At follow-up, exposed workers, who remained at the same workplace no longer had significantly more frequent chronic respiratory symptoms. The prevalence of positive serology was still higher (P < 0.05) in exposed workers after 2 years, but at the 6 year follow-up investigation, the difference was no longer significant. Prevalence of positive skin tests after 2 years no longer differed from that of the non-exposed workers. Slopes (ml/years) of the graphs of forced vital capacity (FVC) and one-second forced expiratory volume (FEV1) during the 6 year follow-up after the first investigation did not differ between exposed and non-exposed workers, both before and after adjustment for age, smoking habits and results of skin tests and serology before modification. Before modification, exposed workers with a positive late skin test had a lower FVC than non-exposed workers. After 6 years of follow-up their FVC was still lower, but the difference was no longer significant. There were no indications of selective loss to follow-up. After remedial actions no new cases of humidifier disease occurred. Moreover, the yearly incidence of work-disability in this plant because of chronic respiratory disease, decreased from 1.30% to 0.27% compared with a decrease from 0.30% to 0. 12% in other synthetic fibre plants. CONCLUSION: Follow-up investigation of exposure-effects demonstrated the effectiveness of remedial actions taken against a contaminated humidification system. PMID- 11007341 TI - Homozygous gene deletions of the glutathione S-transferases M1 and T1 are associated with thimerosal sensitization. AB - OBJECTIVE: Thimerosal is an important preservative in vaccines and ophthalmologic preparations. The substance is known to be a type IV sensitizing agent. High sensitization rates were observed in contact-allergic patients and in health care workers who had been exposed to thimerosal-preserved vaccines. There is evidence for the involvement of the glutathione system in the metabolism of thimerosal or its decomposition products (organomercury alkyl compounds). Thus detoxification by polymorphically expressed glutathione S-transferases such as GSTT1 and GSTM1 might have a protective effect against sensitization by these substances. METHODS: To address this question, a case control study was conducted, including 91 Central European individuals with a positive patch-test reaction to thimerosal. This population was compared with 169 healthy controls and additionally with 114 individuals affected by an allergy against para-substituted aryl compounds. The latter population was included in order to test whether possible associations were due to substance-specific effects, or were a general feature connected with type IV immunological diseases. Homozygous deletions of GSTT1 and GSTM1 were determined by polymerase chain reaction. RESULTS: Glutathione S-transferase M1 deficiency was significantly more frequent among patients sensitized to thimerosal (65.9%, P = 0.013) compared with the healthy control group (49.1%) and the "para-compound" group (48%, P = 0.034). Glutathione S-transferase T1 deficiency in the thimerosal/mercury group (19.8%) was barely elevated versus healthy controls (16.0%) and the "para-compound" group (14.0%). The combined deletion (GSTT1-/GSTM1-) was markedly more frequent among thimerosal sensitized patients than in healthy controls (17.6% vs. 6.5%, P = 0.0093) and in the "para-compound" group (17.6% vs. 6.1%, P =0.014), revealing a synergistic effect of these enzyme deficiencies (healthy controls vs. thimerosal GSTM1 negative individuals, OR = 2.0 [CI = 1.2-3.4], GSTT1-, OR = 1.2 [CI = 0.70-2.1], GSTM1/T1-, OR = 3.1 [CI = 1.4-6.5]). CONCLUSIONS: Since the glutathione-dependent system was repeatedly shown to be involved in the metabolism of thimerosal decomposition products, the observed association may be of functional relevance. PMID- 11007342 TI - Comparison between blood and urinary toluene as biomarkers of exposure to toluene. AB - OBJECTIVES: To compare blood toluene (TOL-B) and urinary toluene (TOL-U) as biomarkers of occupational exposure to toluene, and to set a suitable procedure for collection and handling of specimens. METHOD: An assay based on headspace solid-phase microextraction (SPME) was used both for the determination of toluene urine/air partition coefficient (lambdaurine/air) and for the biological monitoring of exposure to toluene in 31 workers (group A) and in 116 non occupationally exposed subjects (group B). Environmental toluene (TOL-A) was sampled during the work shift (group A) or during the 24 h before specimen collection (group B). Blood and urine specimens were collected at the end of the shift (group A) or in the morning (group B) and toluene was measured. RESULTS: Toluene lambdaurine/air was 3.3 +/- 0.9. Based on the specimen/air partition coefficient, it was calculated that the vial in which the sample is collected had to be filled up to 85% of its volume with urine and 50% with blood in order to limit the loss of toluene in the air above the specimen to less than 5%. Environmental and biological monitoring of workers showed that the median personal exposure to toluene (TOL-A) during the work-shift was 80 mg/m3, the corresponding TOL-B was 82 microg/l and TOL-U was 13 microg/l. Personal exposure to toluene in environmentally exposed subjects was 0.05 mg/m3, TOL-B was 0.36 microg/l and TOL-U was 0.20 microg/l. A significant correlation (P < 0.05) was observed between TOL-B or TOL-U and TOL-A (Pearson's r = 0.782 and 0.754) in workers, but not in controls. A significant correlation was found between TOL-U and TOL-B both in workers and in controls (r = 0.845 and 0.681). CONCLUSION: The comparative evaluation of TOL-B and TOL-U showed that they can be considered to be equivalent biomarkers as regards their capacity to distinguish workers and controls and to correlate with exposure. However, considering that TOL-U does not require an invasive specimen collection, it appears to be a more convenient tool for the biological monitoring of exposure to toluene. PMID- 11007343 TI - Study of some immunological parameters in workers occupationally exposed to benzene. AB - OBJECTIVE: The aim of the study was to investigate the immunotoxicity of benzene exposure, to establish the correlation between the exposure biomarkers and some immunological parameters, and to assess the possible influence of confounding factors on the results of immunological assay applicable in routine medical surveillance of benzene-exposed workers. METHODS: Forty-nine female workers in the shoemaking industry who were exposed to solvent mixtures and 27 nonexposed controls were examined. Workers were exposed to benzene concentrations of up to 15 ppm, and to toluene of up to 50 ppm. RESULTS: Significant differences in the levels of benzene and toluene in blood and phenols in post-shift urine between the exposed and the control group confirmed solvent exposure. The number of B lymphocytes (P = 0.01) was lower in the shoe workers than in the controls. Significant correlation was found between the level of immunoglobulin G and benzene in the work atmosphere, while confounding factors had no impact on immunological values. CONCLUSION: According to these results, exposure to benzene concentration lower than 15 ppm can induce depression of the circulating B lymphocyte level and therefore this fact could be used to develop a promising method for health surveillance of benzene-exposed workers. However, considerably more effort in the research on benzene immunotoxicity, especially in the search for suitable health surveillance methods, is still required. PMID- 11007344 TI - Experimental exposure to methylformate and its neurobehavioral effects. AB - OBJECTIVES: The aim of the study was to investigate the acute effects of experimental methylformate exposure on the nervous system. METHODS: In an exposure chamber, 20 subjects were exposed to methylformate at 100 ppm [Swiss maximum allowable concentration (MAC)] for 8 h. The same number of subjects with the same ages (between 20 and 30 years), gender and education level (university) were examined by the same procedure as a control group. The subjects did not know if they were exposed or not. Three times (morning, noon, evening) during these 8 h, mood [Profile of Mood States (POMS)], neurobehavioral performance (reaction, Stroop, nonverbal learning, determination, tracking; Wiener Test System), vision (visual acuity, contrast sensitivity, color sensitivity) and postural sway were tested. During an undemanding test (POMS) and a demanding performance task (determination test), pulse, electromyography (EMG) of the forehead and of the neck were recorded. In the morning and evening spirometry [forced vital capacity (FVC), forced one-second expiration volume (FEV), medium expiration flow (MEF) and peak expiration flow (PEF)] and the odor perception threshold were measured. RESULTS: In the evening, in the exposed group, fatigue was significantly increased and the EMG of the forehead during a demanding task showed a different development during exposure. The other tests showed no significant solvent effect, but 16 of 43 test parameters showed a significant effect of time. CONCLUSIONS: The results of this study indicate a possible effect of methylformate exposure on the subjective feeling of fatigue after 8 h exposure at 100 ppm in young and healthy subjects, without measurable impairment of neurobehavioral performance. We assume that a similar effect in normal work, combined with a heavy workload and shift work, can lead to an impairment of productivity, and increase the risk of accidents. PMID- 11007345 TI - Urinary methanol and formic acid as indicators of occupational exposure to methyl formate. AB - OBJECTIVE: To evaluate the validity of methanol (MeOH) and formic acid (FA) in urine as biological indicators of methyl formate (MF) exposure in experimental and field situations. METHODS: The subjects were 28 foundrymen and two groups of volunteers (20 control and 20 exposed). Exposure assessment of the workers was performed by personal air and biological monitoring. Methyl formate vapour collected on charcoal tube was analysed by gas chromatography. The concentration of MF in the exposure chamber (volunteer-study) was monitored by two independent methods [flame ionisation detection (FID) and Fourier transformation infra-red detection (FTIR)]. Urinary metabolites (MeOH and FA) were analysed separately by headspace gas chromatography. RESULTS: The volunteers exposed to 100 ppm MF vapour at rest for 8 h excreted 3.62 +/- 1.13 mg MeOH/l (mean +/- SD) at the end of the exposure. This was statistically different (P < 0.001) from pre-exposure MeOH excretion (2.15 +/- 0.80 mg/1), or from that of controls (1.69 +/- 0.48 mg/l). The urinary FA excretion was 32.2 +/- 11.3 mg/g creatinine after the exposure, which was statistically different (P < 0.001) from pre-exposure excretion (18.0 +/- 9.3 mg/g creatinine) or that of controls (13.8 +/- 7.9 mg/g creatinine). In foundrymen, the urinary FA excretion after the 8 h workshift exposure to a time weighted average (TWA) concentration of 2 to 156 ppm MF showed a dose-dependent increase best modelled by a polynomial function. The highest urinary FA concentration was 129 mg/g creatinine. The pre-shift urinary FA of the foundrymen (18.3 +/- 5.6 mg/g creatinine) did not differ from that of controls (13.8 +/- 7.9 mg/g creatinine). The urinary MeOH excretion of the foundrymen after the shift, varied from < 1 to 15.4 mg/l, while the correlation with the preceding MF exposure was poor. The foundrymen excreted more (P = 0.01) FA (2.12 +/- 3.56 mg/g creatinine) after the workshift than experimentally, once-exposed volunteers (0.32 +/- 0.11 mg/g creatinine) at a similar inhaled MF level of 1 ppm). CONCLUSIONS: In spite of its high background level in non-exposed subjects, urinary FA seems to be a useful biomarker of methyl formate exposure. The question remains as to what is the reason for the differences in chronic and acute exposure respectively. PMID- 11007346 TI - Skin absorption of some vaporous solvents in volunteers. AB - OBJECTIVES: To determine the dermal absorption rates of vaporous 1,1,1 trichloroethane (111TRI), trichloroethene (TRI), tetrachloroethene (TETRA), hexane (HEX), toluene (TOL) and m-xylene (XYL) in humans. The determined absorption data were used for the validation of two published models for prediction of non-steady-state skin absorption. METHODS: Five volunteers were dermally exposed on an area of about 1,000 cm2 (forearm and hand) for 20 or 30 min. An inhalation exposure with a known dose rate served as a reference. Using the solvent concentrations in exhaled air, measured after both inhalation and dermal exposure, we calculated the maximum absorption rate into the blood, and the average absorption rates into the skin throughout the exposure, using the linear system dynamics method. RESULTS: The absorption rates into the skin, normalised for exposure concentration, amounted to 0.021 cm/h (111TRI), 0.049 cm/h (TRI), 0.054 cm/h (TETRA), 0.013 cm/h (HEX), 0.14 cm/h (TOL), and 0.12 cm/h (XYL). The maximum absorption rates into the blood ranged from 0.005 nmol/h for 111TRI and HEX to 0.050 nmol/hr for TOL. The ratios between the predicted and experimental values of the absorption rates into the skin ranged, for the model of Cleek and Bunge [4], from 0.3 (HEX) to 1.1 (TRI and TETRA), and for the model of Wilschut and Ten Berge [22], from 1.1 (HEX) to 4.7 (XYL). CONCLUSION: The linear system dynamics method allowed us to calculate not only the total amount absorbed by the skin but also the maximum absorption rate into the blood. The steady-state absorption rate, usually described by a permeability constant, will be below the absorption rate into the skin and above the maximum absorption rate into the blood. The skin absorption rates predicted by the models showed a good agreement with the experimental values. A comparison of the estimated whole-body skin uptake with the inhalatory uptake from the same atmosphere, revealed that the dermal uptake contributed from 0.1% (HEX) to 1% (TOL and XYL) to the total uptake. PMID- 11007347 TI - Quaternary ammonium compounds and occupational asthma. AB - OBJECTIVE: Quaternary ammonium compounds, among which benzalkonium chloride is one of the best-known, are commonly used as antiseptics, disinfectants, detergents and preservatives. They can cause occupational asthma, which however, has been rarely reported so far, despite wide use of these products. We report three such cases. Possible mechanisms causing asthma are discussed, taking into account their characteristics. METHODS AND RESULTS: Our patients, all female nurses, manifested asthma symptoms upon handling disinfectant solutions containing benzalkonium chloride. Work-related fall in PEFR was observed in all of them. The diagnosis was confirmed by challenge tests where the patients were exposed, in a closed chamber, to suspected disinfectant contained in a tray. All of the women developed early or delayed symptoms upon exposure. Similar challenge tests to placebo or other disinfectants devoid of quaternary ammonium compound were negative. CONCLUSION: These three cases, in addition to others reported in the literature, point out an as yet poorly known etiology of occupational asthma to quaternary ammonium compounds in hospital employees. The exact mechanism of the action remains unexplained. PMID- 11007349 TI - Web alert. Transplantation cancer. PMID- 11007348 TI - Recommendations for the categorization of germ cell mutagens. AB - Germ cell mutagens are currently classified into three categories in the German List of MAK and BAT Values. These categories have been revised and extended by analogy with the new categories for carcinogenic chemicals. Germ cell mutagens produce heritable gene mutations, and heritable structural and numerical chromosome aberrations in germ cells. The original categories 1 and 2 for germ cell mutagens remain unchanged. Two new categories 3A and 3B are proposed for chemicals suspected to be germ cell mutagens. A new category 5 is proposed for germ cell mutagens with low potency that contribute negligibly to human genetic risk provided the MAK value is observed. PMID- 11007350 TI - The impact of preprint servers and electronic publishing on biomedical research. PMID- 11007351 TI - Xenotransplantation: reappraising the risk of retroviral zoonosis. PMID- 11007352 TI - Chemokines and their receptors in allograft rejection. AB - Despite current recognition of over 40 chemokines and more than 18 chemokine receptors, understanding of their role in transplant immunobiology and transplant rejection is extremely limited and fragmentary. Recent literature has demonstrated the presence of chemokines and their receptors in transplants and some studies demonstrate important functional roles. PMID- 11007353 TI - Mechanisms of chronic rejection. AB - Chronic rejection remains the major obstacle to long-term allograft survival. Detailed understanding of putative etiologic risk factors, both antigen-dependent and -independent, is important for designing effective therapeutic strategies to ameliorate this process. Cell senescence may be an important factor in chronic rejection. PMID- 11007354 TI - The role of T cell apoptosis in transplantation tolerance. AB - Rejection of fully MHC-mismatched allografts entails the direct recognition of donor MHC molecules (direct antigen presentation) and the activation of an unusually large mass of alloreactive T cells. There is compelling evidence that apoptotic cell death of activated T cells is a critical initial step in the induction of peripheral allograft tolerance with regimens that are not inherently lymphoablative and that therapies that block T cell activation and T cell apoptosis also block the acquisition of tolerance. Thus, T cell apoptosis may play an important role in reducing the size of cytopathic T cell clones and this process may also promote the development and expansion of immune regulatory cells that are essential in the maintenance of allograft tolerance. PMID- 11007355 TI - Dendritic cells and prospects for transplantation tolerance. AB - The past year has witnessed the resolution of some long-standing enigmas surrounding the immunobiology of dendritic cells, illuminating their opposing roles in peripheral tolerance and allograft rejection. Nevertheless these advances have posed many new questions, the answers to which may subtly influence our approach to the treatment of rejection while bringing ever closer the prospect of donor-specific transplanation tolerance. PMID- 11007356 TI - Tolerance induction by 'megadose' transplants of CD34+ stem cells: a new option for leukemia patients without an HLA-matched donor. AB - Early studies in murine models and more recent clinical data in heavily pre treated leukemia patients have shown that escalation of the dose of hematopoietic progenitor cells can overcome major genetic barriers and enable rapid and durable engraftment of haploidentical, three-locus-mismatched transplants without graft versus-host disease. In vitro studies suggest that veto cells within the progenitor population most probably mediate this facilitating effect. PMID- 11007357 TI - New strategies for bone marrow transplantation. AB - Adoptive immunotherapy of hematologic malignancies and metastatic solid tumors by donor lymphocyte infusion following induction of host-versus-graft transplantation tolerance against can best be achieved following nonmyeloablative stem-cell transplantation (NST). Induction of mixed chimerism may represent the best approach for induction of transplantation tolerance to donor alloantigens. Thus NST may become the optimal approach for the treatment of nonmalignant diseases, where replacement of host with donor hematopoietic cells is indicated: for correction of genetic or stem cell deficiency diseases; as a platform for immunotherapy of autoimmune or infectious diseases; or for induction of tolerance to organ allografts. PMID- 11007358 TI - Knowledge about transplantation tolerance gained in primates. AB - Research in transplantation tolerance relies on application of successful strategies in a nonhuman-primate organ transplant model for preclinical testing. Three principal approaches are being evaluated: hematopoietic chimerism, lymphocyte depletion and costimulation blockade. Interactive work in these three areas has yielded much new information on mechanisms of tolerance and opened the door to pilot clinical trials. Clinical application of xenotransplantation is likely to depend on such strategies. PMID- 11007359 TI - Pharmacodynamics of immunosuppressive drugs. AB - The inability to measure the effects of immunosuppressive drugs on immune cells in vivo has always severely limited preclinical drug development, the design and interpretation of clinical trials and the optimal clinical use of this drug class in transplantation. Now, new technologies using microliter samples of whole blood and exploiting the specificity, sensitivity and versatility of flow cytometry have been developed. These novel techniques not only are illuminating the 'black box' that has obscured the pharmacodynamic effects of immunosuppressants but also are uncovering new mechanisms of action of these drugs. Pharmacodynamic assays measure biologically relevant events in vivo, since changes in lymphocyte functions in blood collected from immunosuppressed graft recipients faithfully reflect histopathologic events within allograft tissue. PMID- 11007360 TI - Prospects for xenotransplantation. AB - Pig-to-primate organ survival has been extended from a few minutes to weeks and occasionally months, following the development of transgenic pigs that express human complement-regulatory proteins, efficient antibody removal technologies and immunosuppressive strategies. The current limitation to the clinical application of this technology is acute vascular rejection, and an understanding of the mechanisms of this process and the development of modalities to overcome it are key to making significant progress at solving the critical shortage of organs for transplantation. Approaches that address this issue are underway in a number of laboratories. PMID- 11007361 TI - Cytokine-secreting tumor cell vaccines. AB - Modification of the tumor microenvironment with gene transfer techniques stimulates two immune mechanisms that effectuate tumor destruction. One involves improved tumor-antigen presentation for the development of specific cellular and humoral immunity. The second involves compromise of the tumor vasculature by soluble factors and leukocytes. PMID- 11007362 TI - Design and evaluation of antigen-specific vaccination strategies against cancer. AB - After studies in preclinical mouse models, the efficacy and safety of tumor specific vaccination strategies is currently being evaluated in cancer patients. The first wave of clinical trials has shown that in general such vaccination strategies are safe. However examples of clinical responses, especially in conjunction with vaccine-induced immune responses, are still scarce. The fact that most trials have so far been performed with end-stage cancer patients can largely account for this deficit. Greater efficacy of anticancer vaccines is expected in patients with less-progressed disease. In addition, the detection of both natural and vaccine-induced T cell immunity needs further improvement. PMID- 11007363 TI - The dendritic cell and human cancer vaccines. AB - Over the past ten years, the identification of the critical role that dendritic cells (DCs) play in stimulating a specific immune response has led to their use in cancer and HIV therapy. Interesting responses have been reported but the most effective approach and the duration of these responses are still unclear. The quality of DCs, the means by which tumor antigens are delivered to DCs and the problems associated with monitoring the immune response have made individual studies difficult to compare. Much work is still needed to determine the role that DC-based cancer vaccines will have, the most effective way to deliver DCs to patients and the most relevant antigens to provide to DCs. PMID- 11007364 TI - Costimulatory wars: the tumor menace. AB - Advances in our understanding of T cell costimulatory molecules have provided a vast array of novel approaches to tumor immunotherapy. In the past year, combinatorial immunotherapy based on earlier studies of CTLA-4 blockade, the identification of novel B7-family members, the modulation of CD40 to reverse tolerance to tumor-associated antigens and the use of OX40 to enhance antitumor responses of CD4+ T cells have all contributed to the development of more powerful immunomodulatory cancer therapies. PMID- 11007366 TI - The role of peritoneal dialysis as the first-line renal replacement modality. AB - Twenty years after its introduction, peritoneal dialysis (PD) is a well established alternative to hemodialysis (HD) as a modality of renal replacement therapy. Much debate and research is apparent in the literature, comparing hemodialysis and PD as "opposite" modalities and trying to ascertain which modality should be more optimal. In our opinion, HD and PD are two distinct modalities, each with its own advantages and disadvantages. In addition, it is clear that for both HD and PD, rates of technique failure are high, causing patients to transfer between modalities. The question is thus not which modality is best, but rather, which flow-chart of modalities makes best use of the advantages of each modality, while avoiding its disadvantages. In this respect, HD and PD appear to be complementary modalities. The better preservation of residual renal function, lower risk of infection with hepatitis B and C, better outcome after transplantation, preservation of vascular access, and lower costs are arguments to promote PD as a good initial treatment. When PD-related problems arise (adequacy, ultrafiltration, peritonitis, patient burnout), a timely transfer to HD has to be planned. This editorial tries to review arguments supporting the complementary nature of both modalities, and especially the role of PD as the first-line renal replacement therapy. PMID- 11007365 TI - Building better vaccines: how apoptotic cell death can induce inflammation and activate innate and adaptive immunity. AB - The immunological consequences of apoptosis have been hotly debated. Apoptosis was originally described as a set of cellular morphological changes that occur in the absence of inflammation but the term has been redefined on the basis of a set of conserved molecular events that include the activation of caspases. Though the apoptosis occurring during normal development is immunologically bland or even tolerizing, the apoptotic death after viral infection or after the ligation of Fas can trigger powerful innate and adaptive immune responses. The molecular machinery at the nexus of apoptosis and inflammation includes caspase-1 --an activator of IL-1beta and IL-18 - as well as the double-stranded-RNA-dependent protein kinase pathway and RNaseL pathway, which are key effectors of antiviral immunity. New proapoptotic vaccines induce immune responses that may be able to prevent or treat infectious disease and cancer. PMID- 11007367 TI - Treatment of peritonitis: update or new course. PMID- 11007368 TI - Early start to peritoneal dialysis: to start or not to start? PMID- 11007369 TI - Sclerosing encapsulating peritonitis: what approach should be taken with children? PMID- 11007370 TI - Preservation of residual renal function--an important goal. PMID- 11007371 TI - Adult peritoneal dialysis-related peritonitis treatment recommendations: 2000 update. PMID- 11007372 TI - Preliminary evaluation of incremental peritoneal dialysis in 25 patients. AB - BACKGROUND: Incremental dialysis has been suggested for patients with some residual renal function. However, very little published clinical data exist on the feasibility of this schedule. OBJECTIVES: To assess feasibility of incremental dialysis, with regard to its effect, complications, and impact on quality of life. DESIGN: Pilot prospective study, not controlled. SETTING: Nephrology division, public clinical research hospital. PATIENTS: Twenty-five patients (19 men, mean age 61+/-13 years, body weight 63+/-11 kg) began peritoneal dialysis (the first treatment of uremia) with a single nightly exchange lasting 10 hours or 2 daily exchanges over 12 hours according to creatinine clearance and Kt/N. Patients gave informed consent and reported their work activity, degree of rehabilitation, and their quality of life by answering a questionnaire prepared for this purpose. OUTCOME MEASURES: Survival rate, complications related to peritoneal dialysis, and residual renal and peritoneal clearances. RESULTS: During the study period no patient died. Complications related to dialysis were peritonitis (0.41 episodes/year) and exit-site infection (0.32 episodes/year). All patients continued to work with full rehabilitation and considered 1 or 2 exchanges per day less troublesome than 3 or 4. CONCLUSIONS: Incremental dialysis is well accepted by patients and staff. This technique does not involve a high risk of complications and is economical. Therefore incremental dialysis is feasible. PMID- 11007373 TI - Incremental initiation of dialysis: one center's experience over a two-year period. AB - OBJECTIVE: This pilot study describes our center's experience with peritoneal dialysis (PD) over the past 2 years using a "healthy start" dialysis protocol with an incremental approach to prescription management. DESIGN: Nonrandomized, prospective pilot study. SETTING: Single PD unit of a university teaching hospital. PATIENTS: Thirteen PD patients who initiated dialysis at our center from April 1997 to June 1999. METHODS: Patients initiating PD with residual renal Kt/V of 1.0 - 2.0/week were invited to participate. They were given an initial dialysis prescription so that total (residual renal + dialysis) weekly Kt/V exceeded 2.0. The dialysis prescription was "incrementally" increased as residual renal function (RRF) declined. Data collected for all patients included monthly serum chemistries, residual renal weekly Kt/V and creatinine clearance (CCr) at 1 to 2-month intervals, and peritoneal weekly Kt/V and CCr at 3-month intervals and 1 month after each prescription change. RESULTS: To date, we have followed 13 patients on our incremental PD protocol for a total of 159.3 patient-months. Mean serum albumin concentration and mean normalized protein equivalent of nitrogen appearance (nPNA) were stable throughout the study. Mean total Kt/V and CCr remained above the recommended targets of 2.0/wk and 60 L/wk, respectively. Residual renal function declined rather slowly in our PD patients. One patient died from complications of aortic valve surgery and a second died from pneumonia. A third patient died from peritonitis. One patient required a new Tenckhoff catheter after catheter migration. Three patients were temporarily switched to hemodialysis after a hernia repair, a pleural leak, and elective native/transplant nephrectomies, respectively. Two patients were permanently switched to hemodialysis: one after an episode of peritonitis, the second after accidentally damaging his PD catheter. CONCLUSIONS: Providing incremental dialysis to maintain adequate total small solute clearance has been technically feasible in our patient population. However, a larger than expected number of complications was seen in our study. Fortunately, complications were easier to manage due to the presence of RRF. Because this study was not designed to compare outcome with that observed after traditional initiation of dialysis, further large-scale studies are needed. PMID- 11007374 TI - Continuous adaptation of the dialysis prescription maintains adequate Kprt/V in CAPD patients. AB - OBJECTIVE: To analyze the effect of individual adaptation of the dialysis prescription in continuous ambulatory peritoneal dialysis (CAPD), as compensation for the decline of residual renal function (RRF), on peritoneal (Kpt/V) and total (Kprt/V) urea clearance as well as on peritoneal (Kpcr) and total weekly creatinine clearances (CCr). DESIGN: Retrospective analysis of a 2-year period. PATIENTS: We analyzed 18 patients [15 male, 3 female; mean age 58.2 (24 - 80) years]. MAIN OUTCOME MEASURES: Correlations between increased dialysis prescription and Kpt/V, Kprt/V, and Kpcr. Kprt/V and CCr measurements were based on a 24-hour dialysate and urine collection. Measurements were performed over a time period of 3 to 6 months. RESULTS: The results show a linear correlation between Kpt/V and Kpcr and the prescribed volume by kilogram body weight. Kprt/V was increased slightly by increasing the dialysis prescription. Dialysate-to plasma (D/P) ratios of urea and creatinine remained unchanged. The mean prescribed dialysate volume increased from 7.4+/-1.1 L to 10.6+/-2.5 L. Mean Kprt/V could be maintained on a stable level for a 36-month period. CONCLUSION: By adapting the dialysis prescription on an individual basis to the continuous decline of RRF, and taking the patient's body weight into account in the prescription decision, the increases in Kpt/V offset the decline in RRF. We recommend early individualization of prescription for patients on CAPD. PMID- 11007375 TI - Rate of decline of residual renal function in patients on continuous peritoneal dialysis and factors affecting it. AB - OBJECTIVE: We analyzed residual renal function (RRF) in a large number of new peritoneal dialysis (PD) patients to prospectively define the time course of decline of RRF and to evaluate the risk factors assumed to be associated with faster decline. STUDY DESIGN: Single-center, prospective cohort study. SETTING: Home PD unit of a tertiary care University Hospital. PATIENTS: The study included 242 patients starting continuous PD between January 1994 and December 1997, with a minimum follow-up of 6 months and at least three measurements of RRF. MEASUREMENT: All patients had data on demographic and laboratory variables, episodes of peritonitis and the use of aminoglycoside (AG) antibiotics, temporary hemodialysis, and number of radiocontrast studies. Adequacy of PD was measured from 24-hour urine and dialysate collection and peritoneal equilibration test using standard methodology. Further data on RRF was collected every 3 to 4 months until the patient became anuric (urine volume < 100 mL/day or creatinine clearance < 1.0 mL/min) or until the end of study in December 1998. OUTCOME MEASURE: The slope of the decline of residual glomerular filtration rate (GFR) (an average of renal urea and creatinine clearance) was the main outcome measure. Risk factors associated with faster decline were evaluated by a comparative analysis between patients in the highest and the lowest quartiles of the slopes of GFR, and a multivariate analysis using a stepwise option within linear regression and general linear models. RESULTS: There was a gradual deterioration of residual GFR with time on PD, with 40% of patients developing anuria at a mean of 20 months after the initiation of PD. On multivariate analysis, use of a larger volume of dialysate (p = 0.0001), higher rate of peritonitis (p = 0.0005), higher use of AG (p = 0.0006), presence of diabetes mellitus (p = 0.005), larger body mass index (BMI) (p = 0.01), and no use of antihypertensive medications (p = 0.04) independently predicted the steep slope of residual GFR. Male gender, higher grades of left ventricular dysfunction, and higher 24-hour proteinuria were associated with faster decline on univariate analysis only. CONCLUSION: Faster decline of residual GFR corresponds with male gender, large BMI, presence of diabetes mellitus, higher grades of congestive heart failure, and higher 24 hour proteinuria. Higher rate of peritonitis and use of AG for the treatment of peritonitis is also associated independently with faster decline of residual GFR. Whether the type of PD (CAPD vs CCPD/NIPD) is associated with faster decline of residual GFR remains speculative. PMID- 11007376 TI - Residual renal function in a large cohort of peritoneal dialysis patients: change over time, impact on mortality and nutrition. AB - OBJECTIVE: Residual renal function contributes importantly to total solute clearance in peritoneal dialysis (PD) patients. This study was designed to examine the progression of residual renal function over time and its impact on nutrition and mortality in PD patients in the six New England states (ME, NH, VT, CT, MA, RI) comprising End Stage Renal Disease (ESRD) Network 1. DESIGN: As part of the ESRD Clinical Indicators Project, data on 990 PD patients in Network 1 were abstracted from data supplied by dialysis units in the fourth quarter of 1997. This included demographic information; dose of PD in L/day; weekly renal, dialysis, and total Kt/V urea; weekly renal, dialysis, and total creatinine clearance (CCr); serum albumin level; and mortality and transplantation information. Data collection was repeated in the second and fourth quarters of 1998 and in the second quarter of 1999. PATIENTS: 990 PD patients in Network 1. OUTCOME MEASURES: The change in total and renal solute clearances over time, the relationship between renal clearance and mortality, and the relationship between renal clearance and nutritional status, as represented by serum albumin. RESULTS: Over the 2-year period, mean weekly renal Kt/V urea and weekly renal CCr dropped significantly. To examine the effect of residual renal function on mortality, patients were divided into high and low (above and below the median) weekly renal Kt/V urea and weekly renal CCr groups. Patients above the median levels of both weekly renal Kt/V urea and weekly renal CCr had a significantly decreased risk of dying during the observation period, after controlling for age, gender, serum albumin level, and diabetic status [OR for high vs low renal Kt/V urea 0.54 (CI 0.34 - 0.84), OR for high vs low renal CCr 0.61 (CI 0.40 - 0.94)]. The mean weekly renal Kt/V urea was significantly and directly correlated with the mean serum albumin level by Spearman rank correlation (R = 0.133, p < 0.001), as was the mean weekly renal CCr (R = 0.115, p < 0.001). CONCLUSIONS: Residual renal function is an important contributor to total solute clearance in PD patients. Even at low levels it is linked to decreased mortality and better nutritional status. PMID- 11007377 TI - Long-term peritoneal dialysis is a risk factor of sclerosing encapsulating peritonitis for children. AB - OBJECTIVE: Sclerosing encapsulating peritonitis (SEP) is a clinical syndrome with a high mortality rate and is a serious complication of peritoneal dialysis (PD). Peritoneal sclerosis (PS) is a histological diagnosis. PS is usually observed in the peritoneal specimens of patients with SEP. Avoiding SEP is considered to be extremely important for pediatric patients who may require long-term PD. In this study, the characteristics of patients with PS were investigated to determine when to perform peritoneal biopsies and how long PD can be performed safely. DESIGN: A retrospective single-center study. SETTING: Tokyo Metropolitan Kiyose Children's Hospital. PATIENTS: A total of 109 children younger than 16 years have received chronic PD in our unit since 1981. Among these children, 16 patients had been on PD for more than 5 years (mean 7.4+/-2.5 years) from May 1992 to March 1999. Peritoneal biopsies were performed in 14 of the 16 patients, who were divided into two groups based on the histological diagnoses: a PS and a peritoneal fibrosis (PF) group. RESULTS: The 14 patients were on PD for a mean of 7.8+/-2.5 years. There were 8 patients with PS and 6 patients with PF. SEP was observed in 2 patients in the PS group. The risk of PS increased with the duration of PD: 57% (8/14) > 5 years, 80% (4/5) > 8 years, and 100% (3/3) > 10 years. All patients in the PS group showed both peritoneal calcifications on abdominal CT scan and poor ultrafiltration at the time of diagnoses. CONCLUSION: Long-term PD was the important risk factor of SEP. If both peritoneal calcification on abdominal CT scan and poor ultrafiltration are observed in a patient on PD more than 5 years, a peritoneal biopsy should be performed. If PS is detected, PD should be discontinued. PMID- 11007378 TI - Peritoneal accumulation of AGE and peritoneal membrane permeability. AB - BACKGROUND: In continuous ambulatory peritoneal dialysis (CAPD), the peritoneal membrane is continuously exposed to high-glucose-containing dialysis solutions. Abnormally high glucose concentration in the peritoneal cavity may enhance advanced glycosylation end-product (AGE) formation and accumulation in the peritoneum. Increased AGE accumulation in the peritoneum, decreased ultrafiltration volume, and increased peritoneal permeability in long-term dialysis patients have been reported. AIM: The purpose of the study was to evaluate the relation between peritoneal membrane permeability and peritoneal accumulation of AGE. METHODS: Peritoneal membrane permeability was evaluated by peritoneal equilibration test (PET) using dialysis solutions containing 4.25% glucose. Serum, dialysate, and peritoneal tissue levels of AGE were measured by ELISA method using polyclonal anti-AGE antibody. Peritoneal biopsy was performed during peritoneal catheter insertion [new group (group N), n = 18] and removal [long-term group (group LT), n = 10]. Peritoneal catheters were removed due to exit-site infection not extended into the internal cuff (n = 6) and ultrafiltration failure (n = 4) after 51.6+/-31.5 months (13 - 101 months) of dialysis. PET data obtained within 3 months after the initiation of CAPD or before catheter removal were included in this study. Ten patients in group N and 4 patients in group LT were diabetic. Patients in group LT were significantly younger (46.5+/-11.1 years vs 57.5+/-1.3 years) and experienced more episodes of peritonitis (3.5+/-2.1 vs 0.2+/-0.7) than group N. RESULTS: Peritoneal tissue AGE level in group LT was significantly higher than in group N, in both nondiabetic (0.187+/-0.108 U/mg vs 0.093+/-0.08 U/mg of hydroxyproline, p < 0.03) and diabetic patients (0.384+/-0.035 U/mg vs 0.152+/-0.082 U/mg of hydroxyproline, p < 0.03), while serum and dialysate levels did not differ between the groups in both nondiabetic and diabetic patients. Drain volume (2600+/-237 mL vs 2766+/-222 mL, p = 0.07) and D4/D0 glucose (0.229+/-0.066 vs 0.298+/-0.081, p < 0.009) were lower, and D4/P4 creatinine (0.807+/-0.100 vs 0.653+/-0.144, p< 0.0001) and D1/P1 sodium (0.886+/-0.040 vs 0.822+/-0.032, p < 0.0003) were significantly higher in group LT than in group N. On linear regression analysis, AGE level in the peritoneum was directly correlated with duration of CAPD (r = 0.476, p = 0.012), number of peritonitis episodes (r = 0.433, p = 0.0215), D4/P4 creatinine (r = 0.546, p < 0.027), and D1/P1 sodium (r = 0.422, p = 0.0254), and inversely correlated with drain volume (r = 0.432, p = 0.022) and D4/D0 glucose (r = 0.552, p < 0.0023). AGE level in the peritoneal tissue and dialysate were significantly higher in diabetics than in nondiabetics in group LT, while these differences were not found in group N. Serum AGE level did not differ between nondiabetics and diabetics in either group N or group LT. Drain volume and D4/D0 glucose were lower and D4/P4 creatinine and D1/P1 sodium higher in diabetics than in nondiabetics in both groups. CONCLUSION: Peritoneal accumulation of AGE increased with time on CAPD and number of peritonitis episodes, and was directly related with peritoneal permeability. Peritoneal AGE accumulation and peritoneal permeability in diabetic patients were higher than in nondiabetic patients from the beginning of CAPD. PMID- 11007379 TI - Left ventricular hypertrophy and clinical outcome in CAPD patients. AB - OBJECTIVE: To determine the clinical outcome of left ventricular hypertrophy (LVH) (left ventricular wall diastole thickness > or = 1.2 cm) detected by echocardiography in nondiabetic, continuous ambulatory peritoneal dialysis (CAPD) patients without dilated cardiomyopathy. DESIGN: A prospective, descriptive study was conducted between 1 July 1995 and 31 January 1998. Patients were followed up for 24 months. SETTING: Peritoneal dialysis unit in a medical school hospital. PATIENTS AND METHODS: Baseline and yearly echocardiograms were carried out on 66 patients receiving CAPD. Cardiac death was assessed. LVH was correlated with outcome. RESULTS: Of 66 nondiabetic CAPD patients without dilated cardiomyopathy, 20 had a normal echocardiogram (LV wall thickness < 1.2 cm), 21 had mild hypertrophy, and 25 severe hypertrophy (LV wall thickness > 1.4 cm in diastole). In the first two groups, 21% were admitted with congestive heart failure (CHF) after starting dialysis. The 1-year cumulative survival was 85% among those with mild hypertrophy and 91% in the normal group. In the group with severe hypertrophy, 57% were admitted at least once with CHF, and the 1-year cumulative survival was 56%. Eighty-two percent of those who died in the severe group, which accounted for the significantly worse survival (p = 0.003), died from cardiac or cerebrovascular causes, compared with none of those with a normal echocardiogram. CONCLUSIONS: Severe LVH was found in a third of our CAPD patients and was associated with a significantly high cardiovascular morbidity and mortality. PMID- 11007380 TI - Insulin efficacy with a new bicarbonate/lactate peritoneal dialysis solution. PMID- 11007381 TI - Prevalence of hepatitis G virus infection in a hemodialysis and a peritoneal dialysis (CAPD) population. PMID- 11007382 TI - Catheter cuff shaving using a novel technique: a rescue treatment for persistent exit-site infections. PMID- 11007383 TI - Peritoneal permeability of Asian Indian patients does not differ from that of non Indian patients (in Toronto, Canada). PMID- 11007384 TI - Literature. July-August 2000. PMID- 11007385 TI - Breast cancer following breast reduction surgery in Sweden. AB - Women undergoing breast reduction surgery have been reported to be at low subsequent risk of breast cancer, especially when the surgery is performed after age 40. To evaluate the age and time-related patterns of cancer risk following surgical removal of breast tissue, we identified 31,910 women who underwent breast reduction surgery from 1965 to 1993 in Sweden using hospital discharge register data. There were 19,975 women (63 percent) under age 40 at surgery. Linkages with Swedish registries for cancer, death, and emigration were based on unique national registration numbers assigned to each Swedish resident. Cancer incidence was contrasted with that expected in the general population based on age- and calendar year-specific data from the nationwide cancer registry. Overall, 161 incident breast cancers were identified during 238,765 person-years of observation (mean, 7.5 years) compared with 223.9 expected (standardized incidence ratio = 0.72; 95 percent confidence interval = 0.61 to 0.84). The reduction in risk of breast cancer was most pronounced for women whose operations were performed after age 50 (SIR = 0.57) and for those followed for more than 5 years (SIR = 0.68). Among women operated on before age 40, risk was nonsignificantly elevated within the first 5 years after surgery (SIR = 1.47; 95 percent CI = 0.89 to 2.30) but tended to be reduced thereafter (SIR = 0.80; 95 percent CI = 0.55 to 1.13). The magnitude of the reduction in risk thus appears directly related to age at surgery. Women followed for an average of 7.5 years after bilateral breast reduction surgery, were at a statistically significant 28 percent decreased risk of breast cancer. The current study is thus consistent with a protective effect following partial removal of breast glandular tissue. PMID- 11007386 TI - Modified autogenous latissimus breast reconstruction and the box top nipple. AB - During the past 3 years, the authors have been using the modified autogenous latissimus myocutaneous flap (MALF) for breast reconstruction in increasing numbers because of occasional patient and surgeon dissatisfaction with other methods of breast reconstruction. They have found this method to have unprecedented reliability, making it preferable to other forms of reconstruction in many patients. Considering the very low morbidity, the high patient satisfaction, and current economic factors, the authors are strong advocates of this form of reconstruction. A procedural outline proposed by McCraw and coworkers is followed, with some useful modifications. An elliptical transverse skin paddle is centered over the back fat roll. The area of the skin ellipse measures approximately 8 +/- 2 cm vertically and 30 +/- 5 cm transversely. After making the skin incision, a feathering technique is used in all directions through the fatty layer overlying the latissimus and in the tissue beyond the anteroposterior borders of the latissimus (not beyond 5 cm from the skin incision). By means of feathering, the shape of a breast mound can be created in the allowable tissue supported by the latissimus. A 180-degree rotation of the flap allows dependentvenous drainage and more bulk in the inferior outer quadrant, where it is needed. In the current series of 47 modified autogenous latissimus breast reconstructions, seromas were common. Other complications included one wound infection, one ulnar neuropraxia, and one fat necrosis. There were no flap necroses (partial or complete) or hematomas. The rarity of complications supports the use of this technique in selected patients. An innovative new technique for nipple reconstruction is also described. The "box top technique" of nipple reconstruction consists of four deepithelialized local flaps covered with a skin graft from the groin. PMID- 11007387 TI - Determinants of patient satisfaction in postmastectomy breast reconstruction. AB - In today's increasingly competitive health care marketplace, consumer satisfaction has become an important measure of quality. Furthermore, measures of satisfaction with treatment inteerventions are influential factors in determining patients' and payers' choices of health care. This study sought to evaluate satisfaction with postmastectomy breast reconstruction and to assess the effects of procedure type and timing on patient satisfaction. As part of the Michigan Breast Reconstruction Outcome Study, patients undergoing first-time mastectomy reconstruction were prospectively evaluated, including cohorts of women choosing expander/implant, pedicle TRFAM flap, and free TRAM flap procedures. Preoperatively and 1 year postoperatively, participants completed a questionnaire that collected a variety of health status information. The postoperative questionnaire had an additional seven items assessing both general satisfaction with reconstruction (five items) and aesthetic satisfaction (two items) as separate subscales. Patients were asked to respond to each item using a five point Likert scale. Item responses ranged from 1, indicating high satisfaction, to 5, reflecting low satisfaction. In the data analysis, only patients responding with a 1 or 2 for all of the items within a subscale were classified as "satisfied" for the subscale. To assess the effects of procedure type (implant, pedicle TRAM flap, and free TRAM flap) and timing (immediate versus delayed) on satisfaction and to control for possible confounding effects from other independent variables, multiple logistic regression was employed. In our analysis, odds ratios and associated 95 percent confidence intervals were calculated for each independent variable in the regression. Statistical significance was designated at the p < or = 0.05 level. A total of 212 patients were followed during the period of 1994 to 1997, including 141 immediate and 71 delayed reconstructions. The study population consisted of 49 expander/implant, 102 pedicle TRAM flap, and 61 free TRAM flap reconstruction patients. The analysis showed a significant association between procedure type and patient satisfaction. TRAM flap patients (both free and pedicle) appeared to have significantly greater general and aesthetic satisfaction compared with expander/implant patients (p = 0.03 and 0.001, respectively). Furthermore, pedicle TRAM flap patients were more aesthetically satisfied than those with free TRAM flaps (p = 0.072). The other independent variables of age and procedure timing did not appear to significantly affect either general or aesthetic satisfaction. However, preoperative physical activity was positively correlated with general satisfaction at the p = 0.034 level. The choice of procedure seems to have a significant effect on both aesthetic and general patient satisfaction with breast reconstruction. In this study, autogenous tissue reconstructions produced higher levels of patient aesthetic and general satisfaction compared with implant techniques. Pedicle and free TRAM flap patients do not seem to differ significantly in general satisfaction. However, women receiving pedicle TRAM flaps reported greater aesthetic satisfaction compared with patients undergoing free TRAM flaps. Furthermore, patient age and procedure timing may not have an effect on patient satisfaction with breast reconstruction. PMID- 11007388 TI - A new technique of scarless expanded forehead flap for reconstructive surgery. AB - The forehead flap is an ideal flap for reconstructive surgery, especially for that involving reconstruction of the face and neck. However, it is usually limited to use in nasal reconstruction, even when performed in conjunction with tissue expansion, because of the severe visible morbidity of the donor site. In this article, the author discusses his development of a new technique of forehead flap, performed in conjunction with tissue expansion, for reconstructive surgery without visible scarring at the donor site. The technique involved positioning a tissue expander in the forehead pocket under the occipitofrontal muscle and serially inflating the expander over a period of approximately 4 to 6 weeks. Thereafter, an expanded forehead flap was created from the frontal hairline area on the basis of the location of the superficial temporal vessels and transferred into 16 recipient sites in 13 patients as an island flap (n = 8), a free flap (n = 1), or a local random flap (n = 7). The donor site was closed directly into the frontal hairline, without any visible scar. With the author's experience in the use of the island flap for nasal, facial, and neck reconstruction and of the free flap for reconstruction in the extremities, the flap could be as large as 8 x 18 cm without inducing flap necrosis or problems with donor-site closure. All patients (n = 13) had acceptable donor-site aesthetic results, without visible scarring. The results indicate that the flap could be a safe, ample, and color matched flap for reconstruction of the face and neck and could also diminish donor-site morbidity to a minimum, without an unsightly visible scar. Furthermore, the flap could be formed into a customized free flap, with the above mentioned advantages, to be transferred to any part of the body. PMID- 11007389 TI - Ten-year review of a prospective randomized controlled trial of textured versus smooth subglandular silicone gel breast implants. AB - Although textured silicone breast implants have been shown to reduce the incidence of capsular contracture, there is little evidence if this effect is maintained in the long term. It has been 10 years since the double-blind randomized trial in which 53 patients received either Mentor smooth (26) or textured silicone gel implants (27). Of the 14 patients who were not known to have developed a contracture in the smooth group, 11 were reviewed. Three had bilateral contractures. In the textured group, 18 of the 24 patients not known to have contractures were reviewed. None had developed contractures. At 10 years, the incidence of capsular contracture was 65 percent of patients with smooth implants (an increase of 6 percent on the 3-year results) and 11 percent for the textured implant patients (no change on the 3-year results). A database containing the details of 1100 patients reinforces these results by examining the differences in contracture rates of textured, smooth, and polyurethane-coated implants. The effect of submuscular placement on reducing contracture rates regardless of texturing is discussed, as is the apparent increase in capsular contracture in patients who smoke. PMID- 11007390 TI - Recurrence of subglandular breast implant capsular contracture: anterior versus total capsulectomy. AB - The objective of this study was to determine whether the type of capsulectomy, anterior or total, affects the recurrence of capsular contracture around subglandular silicone-gel breast implants. A retrospective analysis was performed of patients who underwent either anterior or total capsulectomy for Baker grade 3 or 4 subglandular capsular contracture in our unit. All patients were invited to a review clinic where their capsular status was assessed. There were 100 anterior disc capsulectomies in 60 patients between 1988 and 1997 and 99 total capsulectomies in 60 patients between 1990 and 1998. The follow-up in the former group was a median of 7 years and mean 6.9 years, compared with median 2.5 and mean 3.1 years in the latter group. Eighty-six percent of the implants removed from both groups at capsulectomy were smooth-walled gel-filled implants. Sixty nine breasts in the anterior group received textured gel implants at capsulectomy; the remaining 31 received polyurethane-coated Meme implants. In the total capsulectomy group, all but two breasts (one patient) received textured gel implants. After review, the capsular status was known in 80 percent of the anterior and 92 percent of the total capsulectomy group. The review clinic found eight new contractures in five patients to have developed in the anterior compared with none in the total group. Recurrent contractures affected 50 percent of patients (46 percent of breasts) in the anterior and 11 percent of patients (10 percent of breasts) in the total capsulectomy group. Kaplan-Meier survival analysis was applied to the data. By including only patients who received textured gel implants at capsulectomy, the Logrank found a statistical difference between the two treatment groups (0.01 < p < 0.5). We believe that this study provides some evidence that total capsulectomy for subglandular silicone breast implant capsular contracture results in a lower capsular recurrence than anterior disc capsulectomy. The pattern and risk of recurrence after total capsulectomy and exchange for a modern textured prosthesis appear to approach those following primary augmentation. PMID- 11007391 TI - Lip reconstruction following Mohs' surgery: the role for composite resection and primary closure. AB - Surgical outcomes and patient satisfaction with composite resection and primary closure for the management of upper-lip defects following Mohs' surgery were evaluated. Twenty-seven patients underwent upper-lip reconstruction following Mohs' surgery from 1993 to 1997. Twelve of these patients were selected for this report based on adequate follow-up examinations and photographs. There were nine women and three men with a mean age of 46 years (range, 33 to 70 years). Eleven patients underwent Mohs' surgery for basal cell carcinoma and one patient for squamous cell carcinoma of the upper lip. The defects varied in size and location, often extending beyond a single aesthetic subunit. The reconstruction was performed an average of 7 days after Mohs' surgery (range, 1 to 23 days). In 50 percent of the cases, a full-thickness excision was performed, which included orbicularis oris and inner-lip mucosa. The functional results were graded as near normal to normal in all cases. There were no observed changes in oral continence, eating or speech. Two patients experienced numbness medial to the operative site, but this had no adverse affect on lip function. The aesthetic results were graded as very good to excellent in all cases. Eleven of the 12 patients were satisfied with their lip appearance and function. Conventional wisdom dictates that during reconstruction of upper-lip defects, one should attempt to maintain a majority of the uninvolved tissue for the best result. Although these techniques result in wound closure, they fail to consider lip aesthetics. By using a vertically oriented composite resection of the tipper lip with the additional resection of uninvolved tissue, normal lip architecture is maintained. In our experience, this results in a superior aesthetic and functional result. PMID- 11007392 TI - Abdominal wall closure after intestinal transplantation. AB - Successful surgical closure of the abdominal wall after either combined or isolated intestinal transplantation may present a challenging dilemma for the plastic and reconstructive surgeon because of the following factors: restricted volume of the recipient abdominal cavity; donor-recipient size discrepancies as expressed by the donor to recipient weight ratio; and significant intraoperative edema. The purpose of this investigation is to present clinical experience with 51 consecutive patients who underwent a total of 57 sequential intestinal transplantations at the University of Miami-Jackson Memorial Hospital. A retrospective chart review of 36 pediatric (63 percent) and 21 adult (37 percent) transplantations was performed. Age of the pediatric population ranged from 1 month to 13 years (mean, 2.4 years) and of the adult population from 22 to 55 years (mean, 33.5 years). Several diagnostic classifications necessitated organ transplantation. Because of insufficient donor graft size for the recipient abdominal cavity in 19 transplantations (33 percent), several technical modifications were used to achieve anatomic and functional abdominal wall closure in all patients. In summary, the plastic and reconstructive surgeon should have a significant role in the comprehensive planning and management of abdominal wall closure in this challenging group of patients. PMID- 11007393 TI - Treatment of diabetic neuropathy by decompression of the posterior tibial nerve. AB - A series of 58 operations on 36 patients were performed for decompression of the posterior tibial nerve for the treatment of diabetic neuropathy. Preoperative symptoms included lack of sensation, pain, or both. Eleven of the 36 patients had neurotrophic ulcers, which were treated simultaneously. The operation was found to be effective for relief of pain in 24 of the 28 patients with that complaint (86 percent). Restoration of sensation was less consistent with improvement noted in 18 of the 36 patients (50 percent). The follow-up period ranged from 12 to 84 months (mean, 32 months) and five patients had some degree of recurrent symptoms. No patient has developed a new ulcer after nerve decompression. Wound complications were minimal (12 percent), even though ulcers were treated simultaneously. No patient required surgical treatment for the decompression incision, although one did require hospital admission for treatment of a wound infection. In general, the procedure seemed to be a worthwhile treatment, which should be considered ill selected diabetics with symptomatic neuropathy. PMID- 11007394 TI - Results of decompression of peripheral nerves in diabetics: a prospective, blinded study. AB - Diabetic neuropathy traditionally is considered progressive and irreversible and will result in lower extremity ulceration and amputation in a segment of the diabetic population, despite the best efforts to control serum glucose levels. Restoration of sensation to the diabetic may prevent these complications of neuropathy. The present study was designed to evaluate whether decompression of a peripheral nerve at a known site of anatomic narrowing can restore sensibility to that nerve in the diabetic. Twenty diabetic patients ( 14 type I, 6 type II, with a mean duration of diabetes of 14.8 years) had surgical decompression of a median nerve at the wrist and an ulnar nerve at the elbow, or a decompression of the posterior tibial nerve at the ankle (total of 31 nerves). A therapist, in a manner blind to the operative site, evaluated two-point discrimination in the pulp of the appropriate digit. The postoperative sensibility was compared with that of the nontreated, contralateral extremity. At a mean of 23.3 months, 69 percent of the lower-extremity nerves and 88 percent of the upper-extremity nerves (79 percent overall) had improvement in sensibility. In comparison, 32 percent of the control (not decompressed) contralateral nerves had measurable progression of neuropathy. The hypothesis that decompression of a peripheral nerve in the diabetic will improve sensibility was confirmed at the p < 0.001 level. PMID- 11007395 TI - Forefoot reconstruction by reversed island flaps in diabetic patients. AB - Soft-tissue coverage of the foot is often difficult, especially when the distal third of the foot (dorsal or plantar aspects) is involved. The clinical situation can be further complicated when diabetic patients are affected by painful and unstable wounds of this kind because of the familiar phenomenon of vasculopathy. The purpose of this study was to evaluate the possibility of using distally based foot flaps to cover forefoot defects in diabetic patients. Preoperative selection of patients was the key to this study; those who had other major disease, chronic infection, bone involvement, and/or insufficient foot vascularization were excluded from the study. The authors report a series of 12 diabetic patients in whom the reconstruction of medium-sized defects (ranging from 1.5 x 2.0 cm to 3.0 x 7.0 cm) of the forefoot was performed using distally based dorsalis pedis flaps or medial plantar flaps. The transferred flaps survived and adapted well to the defects, except for one flap in a patient who had a slight venous insufficiency at outset. Wearing their own footwear, patients could walk after 20 to 30 days. After the follow-up period (3 months to 3 years), no skin breakdown in the treated areas was observed. Temporary donor-site pain was reported by medial plantar flap patients, and partial skin graft loss at the donor site occurred in some of the dorsalis pedis patients. The authors suggest that in selected cases, medium-sized soft-tissue defects involving the dorsal aspects or the weight bearing areas of the diabetic foot can be successfully covered with distally based island flaps. PMID- 11007396 TI - A reverse ulnar hypothenar flap for finger reconstruction. AB - A reverse-flow island flap from the hypothenar eminence of the hand was applied in 11 patients to treat palmar skin defects, amputation injuries, or flexion contractures of the little finger. There were three female and eight male patients, and their ages at the time of surgery averaged 46 years. A 3 x 1.5 to 5 X 2 cm fasciocutaneous flap from the ulnar aspect of the hypothenar eminence, which was located over the abductor digiti minimi muscle, was designed and transferred in a retrograde fashion to cover the skin and soft-tissue defects of the little finger. The flap was based on the ulnar palmar digital artery of the little finger and in three patients was sensated by the dorsal branch of the ulnar nerve or by branches of the ulnar palmar digital nerve of the little finger. Follow-up periods averaged 42 months. The postoperative course was uneventful for all patients, and all of the flaps survived without complications. The donor site was closed primarily in all cases, and no patient complained of significant donor-site problems. Satisfactory sensory reinnervation was achieved in patients who underwent sensory flap transfer, as indicated by 5 mm of moving two-point discrimination. A reverse island flap from the hypothenar eminence is easily elevated, contains durable fasciocutaneous structures, and has a good color and texture match to the finger pulp. This flap is a good alternative for reconstruction of palmar skin and soft-tissue defects of the little finger. PMID- 11007397 TI - The boomerang flap in managing injuries of the dorsum of the distal phalanx. AB - Finding an appropriate soft-tissue grafting material to close a wound located over the dorsum of a finger, especially the distal phalanx, can be a difficult task. The boomerang flap mobilized from the dorsum of the proximal phalanx of an adjacent digit can be useful when applied as an island pedicle skin flap. The vascular supply to the skin flap is derived from the retrograde perfusion of the dorsal digital artery. Mobilization and lengthening of the vascular pedicle are achieved by dividing the distal end of the dorsal metacarpal artery at the bifurcation and incorporating two adjacent dorsal digital arteries into one. The boomerang flap was used in seven individuals with injuries involving the dorsal aspect of the distal phalanx over the past year. Skin defects in all patients were combined with bone,joint, or tendon exposure. The authors found that the flap was reliable and technically simple to design and execute. This one-step procedure preserves the proper palmar digital artery to the fingertip and has proven valuable for the coverage of wide and distal defects because it has the advantages of an extended skin paddle and a lengthened vascular pedicle. When conventional local flaps are inadequate, the boomerang flap should be considered for its reliability and low associated morbidity. PMID- 11007398 TI - Buried penis release in adults with suction lipectomy and abdominoplasty. AB - The buried penis in adults who had substantial weight gain was treated with a combination of suction lipectomy of the abdomen and pubic area and the release of the suspensory ligament of the penis and with abdominoplasty with suture anchoring of the pubic and penile skin base dermis to the rectus fascia superiorly. This procedure was performed in 11 patients over the past 10 years with satisfactory results. One patient required additional excess skin excision from the side of the pubic and scrotal area to improve the results. PMID- 11007399 TI - Can dog-ear formation be decreased when an S-shaped skin resection is used instead of a spindle skin resection? A three-dimensional analysis of skin surgery techniques using the finite element method. AB - Dog-ear formation is often unavoidable with resection and suturing of the skin, including spindle excision. Regarding dog-ear formation after basic spindle skin resection during removal of a round tumor of the skin, we quantitatively analyzed the frequency of dog-ear formation with respect to the following three techniques: previous spindle skin resection, S-shaped skin resection, which has been experientially considered to induce limited deformity, and mosque-shaped skin resection for control. To date, by using paper models or sponges, various techniques of skin resection have been simulated in the field of plastic surgery. In the present study, we performed three-dimensional simulation and analyzed three different techniques of skin resection by using the finite element method. As a result, image simulation demonstrated that the frequency of dog-ear formation was limited by S-shaped, spindle, and mosque-shaped skin resection, in descending order. PMID- 11007400 TI - A molecular analysis of the isolated rat posterior frontal and sagittal sutures: differences in gene expression. AB - Although it is one of the most commonly occurring craniofacial congenital disabilities, craniosynostosis (the premature fusion of cranial sutures) is nearly impossible to prevent because the molecular mechanisms that regulate the process of cranial suture fusion remain largely unknown. Recent studies have implicated the dura mater in determining the fate of the overlying cranial suture; however, the molecular biology within the suture itself has not been sufficiently investigated. In the murine model of cranial suture fusion, the posterior frontal suture is programmed to begin fusing by postnatal day 12 in rats (day 25 in mice), reliably completing bony union by postnatal day 22 (day 45 in mice). In contrast, the sagittal suture remains patent throughout the life of the animal. Using this model, this study sought to examine for the first time what differences in gene expression--if any--exist between the two sutures with opposite fates. For each series of experiments, 35 to 40 posterior frontal and sagittal suture complexes were isolated from 6-day-old Sprague-Dawley rat pups. Suture-derived cell cultures were established, and ribonuicleic acid was derived from snap-frozen, isolated suture tissue. Results demonstrated that molecular differences between the posterior frontal and sagittal suture complexes were readily identified in vivo, although these distinctions were lost once the cells comprising the suture complex were cultured in vitro. Hypothetically, this change in gene expression resulted from the loss of the influence of the underlying dura mater. Significant differences in the expression of genes encoding extracellular matrix proteins existed in vivo between the posterior frontal and sagittal sutures. However, the production of the critical, regulatory cytokine transforming growth factor beta-1 was equal between the two suture complexes, lending further support to the hypothesis that dura mater regulates the fate of the overlying cranial suture. PMID- 11007401 TI - Crossover replantation and fillet flap coverage of the stump after ectopic implantation: a case of bilateral leg amputation. AB - A successful case of crossover replantation of the left foot to the stump of the right leg and temporary ectopic implantation of the right amputated foot on the forearm is described. The ectopically implanted right foot was used as a free fillet flap for the late reconstruction of the left leg stump. At the latest follow-up examination, 18 months after the accident, the patient was able to walk independently with a prosthesis on the stump of the left leg. Both the cross replanted foot and the free filleted foot flap, used for the reconstruction of the left leg stump, have maintained adequate protective sensation. The importance of utilization of amputated parts for functional reconstruction is stressed. Crossover replantations and ectopic implantations should be considered in bilateral amputations for the salvage of at least one extremity. PMID- 11007402 TI - Medial plantar flap based distally on the lateral plantar artery to cover a forefoot skin defect. AB - The authors report a simple, single-step procedure to promote the distal transfer of the instep island flap for coverage of the submetatarsal weight-bearing zone. First described in 1991 by Martin et aI, this procedure remained unknown. As opposed to the medial plantar flap, this technique proposes an instep island flap based on the lateral plantar artery. The inflow and outflow of blood is assured by the anastomosis between the dorsalis pedis and lateral plantar vessels. This approach allows for the transfer of similar tissue and provides adequate coverage of the weight-bearing zone of the distal forefoot. PMID- 11007403 TI - Minimally invasive approach to familial multiple lipomatosis. AB - Thirty-five abdominal wall lipomas were removed from a patient with familial multiple lipomatosis using a minimally invasive approach in a cost-effective, reliable, and cosmetically pleasing manner. The surgical technique used is described in this case report. Clinical findings and prior excisions provided the preoperative diagnosis. The abdominal wall was dissected through two small, vertical midline incisions in the suprafascial plane with the aid of a lighted breast retractor. A complete excision of all palpable lipomas was achieved with this approach. The patient had excellent cosmetic results with minimal postoperative scarring. PMID- 11007404 TI - Treatment of finger avulsion injuries with innervated arterialized venous flaps. AB - Complete degloving injury of the digits not amenable to revascularization may leave poor cosmetic and functional results. We used innervated venous flaps from the dorsum of the foot in two patients with traumatic finger degloving injuries. All the flaps successfully provided coverage over the denuded fingers. Good sensation and nearly full rage of motion of the fingers were obtained. There were no donor-site problems. The advantages of this flap are preservation of a major artery of the donor site, easy elevation without deep dissection, and providing a thin, nonbulky tissue and good sensation. The innervated arterialized venous flap is a useful method that provides functional and cosmetic coverage to the severe avulsion injury of the finger. PMID- 11007405 TI - Establishing cleft malformation surgery in developing nations: a model for the new millennium. AB - This three-stage model outlines a safe and effective method for achieving a local cleft board in a developing region. Maintaining local culture and guaranteeing patient safety are paramount concerns. Success is rooted in the constant assessment and recognition of negative forces, including misdirection and stagnation. The key factors are the identification of an interested local host and a source of funding as the site evolves toward independence. As of June 30, 2000, 501 cases had been performed independently and free of charge by the host healthcare provider in Nepal. There had been no major morbidities or mortalities. PMID- 11007406 TI - Facial sensibility testing: a clinical update. PMID- 11007407 TI - Efficacy of preoperative decontamination of the oral cavity. AB - This two-part study consisted of: (1) a survey to assess current practice patterns, and (2) an experiment designed to assess the results of varying intraoral preparations. A 48 percent response rate was obtained from the survey of the 318 active members of the American Society of Maxillofacial Surgeons. This survey demonstrated that significant controversy continues regarding the efficacy of preoperative decontamination of the oral cavity. This prospective, randomized experimental study of 30 patients (saline, 10; povidone-iodine, 10; no preparation, 10) demonstrates a significant and sustained reduction of both anaerobic and aerobic bacteria after intraoral preparation with povidone-iodine solution but not saline. The patient's age, use of preoperative intravenous cephalosporin, and type and length of procedure did not influence the postoperative bacterial counts. Although none of the patients in this study developed an infection, recommendations are provided for standardized preoperative treatment of the oral cavity. PMID- 11007408 TI - The FDA approves saline-filled breast implants: what does this mean for our patients? PMID- 11007409 TI - Management of the bulbous nose. AB - "Bulbous nose" is a term patients often use to describe a "ball" on the end of their nose. This ball can be caused by the abnormal anatomy of alar cartilage or by the overlying soft-tissue coverage. The purpose of this article is to analyze the different causes of bulbous noses and their treatment options. An analysis was done based on four decades of experience and long-term follow-up. We included 10 patients for our discussion. The relationship of the tip to the vault must be analyzed, because it can create optical illusions. For instance, a low bridge makes the tip appear larger; therefore, a bulbous nose may be relative. Similarly, excessive narrowing of the nasal base by alar wedges makes the tip appear wider. Intrinsic causes of a bulbous nose include skin, subcutaneous tissue (including the nasal superficial musculoaponeurotic system, ligaments, and fat), and the shape and direction of the individual crus. Nasal skin varies as to volume and ability to contract; therefore, the shape, direction, or divergence of the individual crura cannot undergo unlimited modifications. There are several surgical possibilities for a given problem. Making the diagnosis of the underlying abnormal anatomy is the most important step; then the most appropriate operation can be selected. Struts, sutures, resection, dome division, and/or dorsal augmentation are all viable options for the management of the bulbous nose. PMID- 11007410 TI - The use of resorbable spacers for nasal spreader grafts. AB - The concept and technique of the use of resorbable synthetic material for nasal spreader grafts are presented. The material is felt to be particularly useful in revision rhinoplasty, in which the likelihood of internal valve collapse is high and the septum (the most common source of material for spreader grafts) often has already been harvested. The material used is a commercially available polymer of polylactic and polyglycolic acid, Lactosorb. It is supplied as a mesh sheet that can be cut to an appropriate size for spreader grafts. Although the material absorbs after approximately 12 months, it is believed that this is sufficient time for the upper lateral cartilages to be stabilized by fibrosis in their new position and to maintain the appropriate internal valve angle. This material was used on 10 patients with valvular collapse undergoing secondary rhinoplasty. In follow-up observations ranging from 12 to 18 months, there has been no recurrence of airway obstruction. PMID- 11007411 TI - In situ split costal cartilage graft harvesting through a small incision using a gouge. AB - A costal cartilage graft is one of the most useful materials in reconstructive plastic surgery. In this article, a technique of in situ split costal cartilage graft harvesting through a small incision (2 to 3 cm) using a gouge is described. The technique used has many advantages: it is a simple technique, is easy to learn, and can be performed quickly through a small incision. By avoiding complete costal cartilage graft harvesting, the associated potential complications such as pleural perforation, chest wall deformities, long-lasting postoperative pain, and incisional scar length are reduced. This technique will be useful in selected cases for which a complete block of costal cartilage graft is not needed. PMID- 11007412 TI - Surgical treatment of the senile upper lip. AB - Aesthetic changes in the aging upper lip constitute a troublesome problem for modern women. During the process of aging, the following alterations appear in the upper lip: (1) vertical wrinkles, (2) reduction in height of the vermilion border along with lengthening of the skin area of the lip, and (3) "disappearance" of the Cupid's bow. In 1993, Guerrissi and Sanchez described a surgical technique that allowed them to correct the effects of these senile changes in 19 patients. With the use of this surgical technique, a strip of skin on the vermilion border was deepithelialized. The remaining dermal flap was buried in the pocket, which was performed by undermining the superior third of the skin of the upper lip. The short-term results were satisfactory, although a slight reduction in height of the vermilion border and a decrease in the thickness of the lip were observed in five patients (26 percent) 4 years postoperatively. Beginning in 1994, the authors began using a new approach combining dermal flap reshaping with simultaneous lip augmentation using dermal fat grafts, Gore-Tex (W. L. Gore and Associates, Flagstaff, Ariz.) or AlloDerm (LifeCell Corp., Branchburg, NJ.). No serious or definitive complications were observed. Scars on the vermilion border were not conspicuous. A peel was necessary at the same time for complete elimination of rhytids. With this method, both the patients and the surgeons were satisfied with the results. PMID- 11007413 TI - Privacy: wanted or unwanted? PMID- 11007414 TI - Rejuvenation of the midface. PMID- 11007415 TI - Facial palsy after abdominal liposuction. PMID- 11007416 TI - Hospital-acquired pressure ulcers: an epidemic. PMID- 11007417 TI - Reducing the hazard of burns and bovie pads. PMID- 11007418 TI - Dermoid cyst on the auriculotemporal area. PMID- 11007419 TI - A surgical technique for the correction of all types of prominent ears? PMID- 11007420 TI - Frontal sinus expansion from an uncommon cause. PMID- 11007421 TI - Intranasal Kirschner Wire Splinting of nasal fractures. PMID- 11007422 TI - Alloderm lip augmentation. PMID- 11007423 TI - New problems in precertifying breast reduction surgery. PMID- 11007424 TI - Desmoid tumor of the hand. PMID- 11007425 TI - The development of the distally based radial forearm flap in hand reconstruction with preservation of the radial artery. PMID- 11007426 TI - Preputial neurofibroma with hypospadias. PMID- 11007427 TI - A nonanimal model for microsurgical training with adventitial stripping. PMID- 11007428 TI - Plastic surgery at the turn of the century. PMID- 11007429 TI - Primum non nocere. PMID- 11007431 TI - W.I.N.: what's in a name? PMID- 11007430 TI - Patient selection in cosmetic surgery. PMID- 11007432 TI - New approaches to molecular profiling of tissue samples. PMID- 11007433 TI - Reconstruction of axial tomographic high resolution data from confocal fluorescence microscopy: a method for improving 3D FISH images. AB - Fluorescent confocal laser scanning microscopy allows an improved imaging of microscopic objects in three dimensions. However, the resolution along the axial direction is three times worse than the resolution in lateral directions. A method to overcome this axial limitation is tilting the object under the microscope, in a way that the direction of the optical axis points into different directions relative to the sample. A new technique for a simultaneous reconstruction from a number of such axial tomographic confocal data sets was developed and used for high resolution reconstruction of 3D-data both from experimental and virtual microscopic data sets. The reconstructed images have a highly improved 3D resolution, which is comparable to the lateral resolution of a single deconvolved data set. Axial tomographic imaging in combination with simultaneous data reconstruction also opens the possibility for a more precise quantification of 3D data. The color images of this publication can be accessed from http://www.esacp.org/acp/2000/20-1/heintzmann.++ +htm. At this web address an interactive 3D viewer is additionally provided for browsing the 3D data. This java applet displays three orthogonal slices of the data set which are dynamically updated by user mouse clicks or keystrokes. PMID- 11007434 TI - Heterogeneous chromosomal aberrations in intraductal breast lesions adjacent to invasive carcinoma. AB - There is evidence that breast cancer is a heterogeneous disease phenotypically as well as molecular biologically. So far, heterogeneity on the molecular biological level has not been investigated in potential precursor lesions, such as ductal hyperplasia (DH) and ductal carcinoma in situ (DCIS). In this study we applied comparative genomic hybridization (CGH) to formalin-fixed, paraffin-embedded breast tissue with DH and DCIS, adjacent to invasive ductal carcinoma (IDC), to screen these potential precursor lesions for whole genomic chromosomal imbalances. Laser-microdissection was used to select pure cell populations from the sections. Isolated DNA was amplified by degenerate oligonucleotide primed PCR (DOP-PCR) and further processed for CGH analysis. Investigating multiple samples (n = 25) from four patients we found an average of 5.6 +/- 0.9 (mean +/- SEM) chromosomal imbalances already present in DH. In the twelve DCIS lesions an average of 10.8 (+/- 0.9) aberrations was identified with 14.8 (+/- 0.8) aberrations in the four adjacent IDC lesions. The increasing number of chromosomal changes in parallel with the histopathological sequence corroborate the hypothesis, that the carcinomas may have developed through a sequential progression from normal to proliferative epithelium and eventually into carcinoma. However, heterogeneous results were identified in the multiple samples per entity from the same patient, demonstrated mainly in the DCIS samples in the chromosomal regions 6p, 9p, 11q, 16p and 17q, in the DH samples by 3p, 16p and 17q. This heterogeneous findings were most pronounced within the DH and was less in the DCIS and IDC samples. The only aberration consistently found in all samples-even in all DH sample-was amplification of the 20q13 region. Our results demonstrate, that the applied combination of laser-microdissection, DOP-PCR and CGH, may serve to analyse breast carcinogenesis pathways in suitable histological material. However, so far, it is unclear how to handle heterogeneous results and these make identification of relevant changes more difficult. Setting a threshold and evaluating only those chromosomal changes which are present in a majority of samples may be one possibility. This involves however, the risk that infrequent but possibly significant aberrations may be missed. Figures on http://www.esacp.org/acp/2000/20-1/aubele. htm. PMID- 11007435 TI - Evaluation of c-erbB-2 overexpression and Her-2/neu gene copy number heterogeneity in Barrett's adenocarcinoma. AB - Amplification of the Her-2/neu gene is accompanied by overexpression of its cell surface receptor product, c-erbB-2 protein. To investigate the degree of intratumoural heterogeneity we applied immunohistochemistry in primary Barrett's adenocarcinoma (BCA) (n = 6) and dysplasia adjacent to the carcinoma (n = 4). In addition, fluorescence in situ hybridisation (FISH) was performed in primary BCA (n = 5) and dysplastic areas (n = 4). For an objective evaluation digital image analysis and laser scanning microscopy were used. Five of six BCA showed a marked intratumoral heterogeneous staining pattern ranging from areas in which the tumour cells were negative or faintly positive to tumour areas with a strong staining of the entire membrane. Among the two dysplastic areas also a heterogeneous staining pattern was observed. FISH analysis revealed marked heterogeneity of intratumoral gene copy number changes in all BCA showing populations with different fractions of cells with polysomy, low level amplification and high level amplification. One dysplasia showed a minor population with Her-2/neu signal clusters. In conclusion, we observed marked intratumoural heterogeneity of c-erbB-2 protein overexpression and Her-2/neu gene copy number in the majority of the primary BCA analyzed. Digital image analysis and laser scanning microscopy were helpful in quantifying the variations in protein expression and DNA copy number in individual tumour cells. The observed heterogeneity could hamper the exact diagnostic determination of the c-erbB-2 status in small biopsies and possibly influence the effectiveness of a potential c-erbB-2 targeting therapy. Figures on http://www.esacp.org/acp/2000/20 1/walch.htm+ ++. PMID- 11007436 TI - Combining dynamic and static robotic telepathology: a report on 184 consecutive cases of frozen sections, histology and cytology. AB - The aim of this paper is to describe the experiments carried out to evaluate the diagnostic efficacy of a dynamic-robotic telepathology system for the delivery of pathology services to distant hospitals. The system provides static/dynamic features and the remote control of a robotized microscope over 4 ISDN lines. For evaluation purposes, 184 consecutive cases of frozen sections (60), gastrointestinal pathology (64), and urinary cytology (60) have been diagnosed at a distance using the system, and the telediagnosis obtained in this way has been compared with the traditional microscopic diagnosis. Diagnostic agreement ranged from 90% in urinary cytology to 100% in frozen sections. The results obtained suggest that such a system can be considered a useful tool for supporting the pathology practice in isolated hospitals. PMID- 11007437 TI - Nuclear DNA content and chromatin pattern of rat rhabdomyosarcoma cell sublines with different metastatic potentials. AB - There is a constant need of features able to characterize potentially metastatic cells among the heterogeneous cell subpopulations which constitute a tumor. Image cytometry of metastatic tumor cells give rise to variable results, partly because of a heterogeneous origin of cells, or potential drug effects. The aim of this work was to characterize nuclear changes observed in metastatic cell clones issued in vitro from the same parental cell population The nuclear phenotypes of 6 cell sublines isolated from a rat rhabdomyosarcoma cell line and differing in their metastatic ability were evaluated by image cytometry on Feulgen-stained preparations. Densitometric [5], geometric [3] and textural [9] features were computed from each nuclear image. For each cell subline, a metastatic score, ranging from 0 to 10, was calculated on the basis of in vivo invasivity data, by measuring the number of pulmonary metastases observed after s.c. graft of tumor cells in rats. Data obtained were compared to karyotype, growth characteristics, and oncogene expressions of cell lines. The nuclear DNA content, the chromosome numbers, the cell sublines doubling times, and the distribution of cells within the cell cycle appear unrelated with this score. On the contrary, increase in metastatic ability is accompanied by changes in chromatin pattern as assessed by textural features. Progressive increase in chromatin condensation can be observed in cell sublines with increasing metastatic score. These results were confirmed by an unsupervised multivariate partitioning of rhabdomyosarcoma cells which identified two separate subsets whose distributions within the analyzed cell lines correlate with their metastatic ability. These data suggest that, in rat rhabdomyosarcoma cell sublines, metastatic ability could be associated with nuclear morphological changes at the level of chromatin texture. PMID- 11007438 TI - Evaluation of tumor heterogeneity of prostate carcinoma by flow- and image DNA cytometry and histopathological grading. AB - BACKGROUND: Heterogeneity of prostate carcinoma is one of the reasons for pretreatment underestimation of tumor aggressiveness. We studied tumor heterogeneity and the probability of finding the highest tumor grade and DNA aneuploidy with relation to the number of biopsies. MATERIAL AND METHODS: Specimens simulating core biopsies from five randomly selected tumor areas from each of 16 Bocking's grade II and 23 grade III prostate carcinomas were analyzed for tumor grade and DNA ploidy by flow- and fluorescence image cytometry (FCM, FICM). Cell cycle composition was measured by FCM. RESULTS: By determination of ploidy and cell cycle composition, morphologically defined tumors can further be subdivided. Heterogeneity of tumor grade and DNA ploidy (FCM) was 54% and 50%. Coexistence of diploid tumor cells in aneuploid specimens represents another form of tumor heterogeneity. The proportion of diploid tumor cells decreased significantly with tumor grade and with increase in the fraction of proliferating cell of the aneuploid tumor part. The probability of estimating the highest tumor grade or aneuploidy increased from 40% for one biopsy to 95% for 5 biopsies studied. By combining the tumor grade with DNA ploidy, the probability of detecting a highly aggressive tumor increased from 40% to 70% and 90% for one and two biopsies, respectively. CONCLUSION: Specimens of the size of core biopsies can be used for evaluation of DNA ploidy and cell cycle composition. Underestimation of aggressiveness of prostate carcinoma due to tumor heterogeneity is minimized by simultaneous study of the tumor grade and DNA ploidy more than by increasing the number of biopsies. The biological significance of coexistent diploid tumor cell in aneuploid lesions remains to be evaluated. PMID- 11007440 TI - EEG bands during wakefulness, slow-wave and paradoxical sleep as a result of principal component analysis in man. AB - Human electroencephalogram (EEG) has been divided in bands established by visual inspection that frequently do not correspond with EEG generators nor with functional meaning of EEG rhythms. Power spectra from wakefulness, stage 2, stage 4 and paradoxical sleep of 8 young adults were submitted to Principal Component Analyses to investigate which frequencies covaried together. Two identical eigenvectors were identified for stage 2 and stage 4: 1 to 8 Hz and 5 to 15 Hz (87.95 and 84.62 % of the total variance respectively). Two eigenvectors were extracted for PS: 1 to 9 Hz and 10 to 15 Hz (81.62% of the total variance). Three eigenvectors were obtained for W: with frequencies between 1 to 7 Hz, 7 to 11 Hz, and 12 to 15 Hz (78.32% of the total variance). Power for all frequencies showed significant differences among vigilance states. These results indicate that slow wave activity can oscillate at higher frequencies, up to 8 Hz, and that spindle oscillations have a wider range down to 5 Hz. No theta band was independently identified, suggesting either that delta and theta oscillations are two rhythms under the same global influence, or that the traditional division of theta band in the human cortical EEG is artificial. Alpha as a band was identified only during wakefulness. Principal component analysis upon spectral densities extracted broad bands different for each vigilance state and from traditional bands, consistent with functional significance of EEG and with frequencies of generators of rhythmic activity obtained in cellular studies in animals. PMID- 11007439 TI - Development of disturbing dreams during adolescence and their relation to anxiety symptoms. AB - STUDY OBJECTIVES: This work assesses the prevalence and development of disturbing dreams among adolescents and the association of these dreams with anxiety. DESIGN: Sex differences in prevalence were analyzed with chi-square analyses. Changes over time were assessed with Wilcoxon tests and cross-tabulation tables. Associations with anxiety and DSM-III-R symptoms were assessed with ANOVA designs. SETTING: N/A. PARTICIPANTS: A total of 610 boys and girls rated their recall of disturbing and normal dreams at both 13 and 16 years of age. Subgroups of subjects were evaluated for anxiety symptoms at age 13 and for DSM-III-R symptoms of separation anxiety (SA), overanxious disorder (OD) and generalized anxiety disorder (GAD) at age 16. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The recall of disturbing dreams was more prevalent for girls than for boys at both ages, and increased over time for girls while it decreased for boys. The recall of normal dreams was also more prevalent for girls at both ages, but this difference could not fully account for the difference in recall of disturbing dreams. Normal dream recall increased from age 13 to 16 for both sexes. The frequent occurrence of disturbing dreams was associated with anxiety at age 13 and with GAD, SA and OD symptoms at age 16 for both sexes. Evidence of more numerous OD symptoms for girls with frequent disturbing dreams suggests that this form of anxiety may partially account for the observed sex difference in disturbing dream prevalence. CONCLUSIONS: The findings highlight a prevalence of disturbing dreams that is especially marked for adolescent girls. Unlike previous cross-sectional studies, which have found the same sex difference, this longitudinal design also calls attention to within-subjects changes in disturbing dream recall. The results also confirm that the frequent recall of disturbing dreams is associated with pathological symptoms of trait anxiety-apparently even as young as 13 years of age. Further study of disturbing dreams may contribute to understanding of associated pathophysiological factors which, too, vary by sex (e.g., PTSD, insomnia, depression). PMID- 11007441 TI - Spectral features of EEG alpha activity in human REM sleep: two variants with different functional roles? AB - Evidence suggests that an important contribution of spectral power in the alpha range is characteristic of human REM sleep. This contribution is, in part, due to the appearance of well-defined bursts of alpha activity not associated with arousals during both tonic and phasic REM fragments. The present study aims at determining if the REM-alpha bursts constitute a different alpha variant from the REM background alpha activity. Since previous findings showed a selective suppression of background alpha activity over occipital regions during phasic REM fragments and, on the other hand, the density of alpha bursts seem to be independent of the presence or absence of rapid eye movements, one expects to find the same spectral power contribution of alpha bursts in tonic and phasic REM fragments. The results indicated that REM-alpha bursts showed a similar power contribution and topographic distribution (maximum energy over occipital regions) both in tonic and phasic REM fragments. This suggests that two variants of alpha activity with different functional roles are present during the human REM sleep: i) background alpha activity, modulated over occipital regions by the presence of rapid eye movements, which may be an electrophysiological correlate of the visual dream contents; and ii) REM-alpha bursts, independent of the presence of rapid eye movements, which could be facilitating the connection between the dreaming brain and the external world, working as a micro-arousal in this brain state. PMID- 11007442 TI - Dynamics of slow-wave activity during the NREM sleep of sleepwalkers and control subjects. AB - STUDY OBJECTIVE: To compare the number and distribution of awakenings from slow wave sleep (SWS) and both the power and dynamics of EEG slow-wave activity (SWA) in sleepwalkers and controls. Somnambulism is considered to be a disorder of arousal from NREM sleep and related to anomalous SWS and SWA. Power spectral analyses have never been used to quantify patients' SWA across sleep cycles. DESIGN: N/A SETTING: N/A PATIENTS: A polysomnographic study was performed on 15 adult sleepwalkers and 15 age- and sex-matched controls. INTERVENTIONS: N/A MEASUREMENTS & RESULTS: Sleepwalkers had a significantly greater number of awakenings from SWS than did control subjects. Controls showed a greater decrease in SWA across NREM cycles. Sleepwalkers had a significantly lower level of SWA during the first NREM period, where most awakenings take place. CONCLUSION: Sleepwalkers appear to suffer from an abnormality in the neural mechanisms responsible for the regulation of SWS. PMID- 11007443 TI - Non-Invasive detection of respiratory effort-related arousals (REras) by a nasal cannula/pressure transducer system. AB - STUDY OBJECTIVES: The published AASM guidelines approve use of a nasal cannula/pressure transducer to detect apneas/hypopneas, but require esophageal manometry for Respiratory Effort-Related Arousals (RERAs). However, esophageal manometry may be poorly tolerated by many subjects. We have shown that the shape of the inspiratory flow signal from a nasal cannula identifies flow limitation and elevated upper-airway resistance. This study tests the hypothesis that detection of flow limitation events using the nasal cannula provides a non invasive means to identify RERAs. DESIGN: N/A. SETTING: N/A. PATIENTS: 10 UARS/OSAS and 5 normal subjects INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: All subjects underwent full NPSG. Two scorers identified events from the nasal cannula signal as apneas, hypopneas, and flow limitation events. Two additional scorers identified events from esophageal manometry. Arousals were scored in a separate pass. Interscorer reliability and intersignal agreement were assessed both without and with regard to arousal. The total number of respiratory events identified by the two scorers of the nasal cannula was similar with an Intraclass Correlation (ICC) =0.96, and was essentially identical to the agreement for the two scorers of esophageal manometry (ICC=0.96). There was good agreement between the number of events detected by the two techniques with a slight bias towards the nasal cannula (4.5 events/hr). There was no statistically significant difference (bias 0.9/hr, 95%CI -0.3-2.0) between the number of nasal cannula flow limitation events terminated by arousal and manometry events terminated by arousal (RERAs). CONCLUSION: The nasal cannula/pressure transducer provides a non invasive reproducible detector of all events in sleep disordered breathing; in particular, it detects the same events as esophageal manometry (RERAs). PMID- 11007445 TI - Supine cephalometric study on sleep-related changes in upper-airway structures in normal subjects. AB - The purpose of this study was to examine sleep-related changes in the upper airway and surrounding structures including the mandible, hyoid bone, and cervical vertebra in normal subjects using lateral cephalograms in the supine position. Eleven male and nine female healthy adults participated in this study. Supine cephalograms were taken at end-expiration during wakefulness and in stage 1-2 non-rapid-eye-movement (NREM) sleep judged from the electroencephalogram and respiratory monitoring. A paired t-test was used to compare differences between the values measured during wakefulness and sleep. A significant increase in the amount of jaw opening was observed during sleep. The anteroposterior width of the upper airway was significantly decreased during sleep. The distances between the hyoid bone and the mandibular symphysis and between the cervical vertebra and the hyoid bone were significantly decreased during sleep. Consequently, the distance between the cervical vertebra and the mandibular symphysis was significantly decreased. These results suggest that the upper-airway dimension is significantly decreased and relatively small structural changes are induced during sleep in normal subjects. PMID- 11007444 TI - Erectile dysfunction in men with obstructive sleep apnea: an early sign of nerve involvement. AB - Erectile dysfunction (ED) is common in men with obstructive sleep apnea (OSAS) but no completely convincing hypotheses about the underlying pathogenic mechanisms have been published in the literature. The aims of the present study were to assess the presence of ED in a group of OSAS patients without daytime respiratory failure and to determine whether this dysfunction was related to peripheral nerve involvement. Evaluation of the bulbocavernosus reflex (BCR) and the somato-sensory evoked potentials of pudendal nerve (PSEPs), the most widely established method of documenting pudendal neuropathies as being the cause of impotence, was performed in 25 patients. Data on BCR were compared with those of 25 healthy males volunteers matched for age. BCR was altered in 17 patients: in 6 it was elicited while in 11 it had a prolonged latency and reduced amplitude. Patients with altered BCR presented an higher AHI, an higher percentage of sleep time spent with SaO2 <90% (TST90) and a lower daytime PaO2. Six patient had clinically silent neurophysiological signs of mild polyneuropathy. The degree of OSAS and gas exchange alteration was more severe in patients with polyneuropathy than in those with isolated BCR alteration. ED is a common finding in OSAS patients and this alteration seems to be related to a nerve dysfunction. The development of nerve dysfunction is associated with a more severe degree of OSAS and nocturnal hypoxia. PMID- 11007447 TI - Voluntary oculomotor performance upon awakening after total sleep deprivation. AB - The potential impact of sleep inertia on measures of voluntary oculomotor control have been surprisingly neglected. The present study examined the effects of 40 hours of sleep deprivation on saccadic (SAC) and smooth pursuit (SP) performance, attentional/visual search performance (Letter Cancellation Task, LCT) and subjective sleepiness (Sleepiness Visual Analog Scale, SVAS) recorded immediately after awakening. Standard polysomnography of nine normal subjects was recorded for 3 nights (1 adaptation, AD; 1 baseline, BSL; 1 recovery, REC); BSL and REC were separated by a period of 40 h of continuous wakefulness, during which subjects were tested every two hours. Within 30 s of each morning awakening, a test battery (SAC, SP, LCT, SVAS) was administered to subjects in bed. For data analysis, mean performance obtained during the day preceding the sleep deprivation night was considered as "Diurnal Baseline" and compared to performance upon awakening from nocturnal sleep. As a consequence of sleep deprivation, SWS percentage was doubled during REC. Saccade latency increased and velocity decreased significantly upon awakening from REC as compared to the other three conditions (Diurnal baseline, AD awakening, BSL awakening); accuracy was unaffected. As regards SP, phase did not show any impairment upon awakening, while velocity gain upon awakening from REC was significantly lower as compared to the other conditions. Finally, number of hits on LCT upon awakening from REC was significantly lower and subjective sleepiness higher as compared to Diurnal Baseline. It is concluded that 40 h of sleep deprivation significantly impaired performance to SAC and SP tasks recorded upon awakening from recovery sleep. This performance worsening is limited to the measures of speed, while both SAC accuracy and SP phase do not show a significant decrease upon awakening. Since saccadic velocity has recently been found to negatively correlate with simulator vehicle crash rates, it is suggested that the adverse effects of sleep deprivation on sleep inertia magnitude should be avoided by any personnel who may have to perform critical tasks involving high oculomotor control immediately after awakening. PMID- 11007446 TI - Effects of sleep loss on waking actigraphy. AB - STUDY OBJECTIVES: To assess the effect of sleep loss and the effect of a sedating drug on waking actigraphy DESIGN: N/A SETTING: N/A PARTICIPANTS: Seventeen healthy volunteers, aged 19-35 yrs INTERVENTIONS: Four night-day treatments presented in a Latin Square Design: placebo-8 hr time-in-bed (TIB), placebo-4 hr TIB, placebo-0 hr TIB, and diphenhydramine 50 mg-8 hr TIB. MEASUREMENTS AND RESULTS: After the appropriate TIB, medication was administered at 09:00 hr, the Multiple Sleep Latency Test at 09:30, 11:30, 13:30, 15:30, and 17:30 hr, and a 45 min performance battery at 10:30, 14:30, and 16:30 hr. Each day the volunteers wore actigraphs from 0700-1800 hrs. Decreasing TIB was associated with decreased daily mean sleep latency on the MSLT with 4 and 0 hrs differing from 8 hrs and each other. Daytime activity also was reduced by the reduced prior TIB. Increased inactivity relative to the 8 hr TIB developed between the 4 hr and 0 hr TIBs, with 4 hrs differing from 0 hrs, but not 8 hrs. Diphenhydramine 50 mg reduced mean daily sleep latency and increased percent inactive time relative to placebo. On the MSLT diphenhydramine was intermediate to 4 hr and 0 hr TIB and on actigraphy it was similar to 0 hr TIB. CONCLUSIONS: The difference in the effect of diphenhydramine on these actigraphy and MSLT may reflect the different sensitivities of the measures. PMID- 11007448 TI - Maintenance of alertness and performance by a brief nap after lunch under prior sleep deficit. AB - We examined the effects of a 15-min nap after lunch on subsequent alertness, performance, and autonomic function following a short sleep the preceding night. Subjects were 12 healthy students who had slept for only 4 hours the night before being tested. They experienced both nap and no-nap conditions in a counterbalanced order, at least a week apart. The nap condition included a 15-min nap opportunity (12:30-12:45) in bed with polygraphic monitoring. We measured the P300 event-related potential, subjective sleepiness (Visual Analog Scale), and electrocardiogram (ECG) at 10:00, 13:15, and 16:15, and task performance (logical reasoning and digit span) at 10:00, 11:30, 13:15, 14:45, 16:15, and 17:45. Mean home sleep measured by actigraphy was 3.5 hours under both conditions. At 13:15, the P300 latency after the nap was significantly shorter than after no nap, but its amplitude was not affected by napping. Subjective sleepiness at 13:15 and 14:45 was significantly lower, and accuracy of logical reasoning at 13:15 was significantly higher after the nap than after no nap. No other performance measures or the ECG R-R interval variability parameters differed significantly between the nap and no-nap conditions. Mean total sleep time during the nap was 10.2 min, and no stage 3 and 4 sleep was observed. The above results suggest that under prior sleep deficit, a 15-min nap during post-lunch rest maintains subsequent alertness and performance, particularly in the mid-afternoon. PMID- 11007449 TI - Automatic auditory information processing in sleep. AB - STUDY OBJECTIVES: The mismatch negativity (MMN) component of the event-related potentials reflects the automatic detection of sound change. Only a few researchers have investigated the MMN elicitation during sleep in adult human and some of them reported that MMN amplitude was decreased in sleep compared to in waking. However, it is not clear that the decrease of MMN amplitude was due to increased drowsiness or long-term response decrement. Two experiments were conducted to clarify whether or not the MMN was elicited in each sleep stage. We presented auditory stimuli to subjects continuously from waking until sleep state (Experiment 1). Using the same experimental condition, we examined whether or not MMN amplitude was influenced by long-term stimulus presentation (80min.) and by vigilance level (Experiment 2). DESIGN: N/A SETTING: N/A PARTICIPANTS: N/A INTERVENTIONS: N/A MEASUREMENTS & RESULTS: Experiment 1: MMN was obtained in both drowsiness and REM sleep. MMN was significantly smaller in amplitude and shorter in latency in the both stages than in the waking state. However, MMN was not found in another sleep stage. Experiment 2: Amplitudes were no different among 0 20 min., 20-40 min., 60-80 min. But it was decreased in 40-60 min. and power value of alpha-wave was decreased in 40-60 min. CONCLUSIONS: To obtain the reliable data, by using the automatic spectral analysis, we confirmed that MMN was elicited in REM sleep. MMN was not influenced by long-term stimulation. The result suggested that auditory stimuli could be processed in the pre-attentive sensory memory even in REM sleep. PMID- 11007450 TI - A possible role for nitric oxide at the sleep/wake interface. AB - Cholinergic neurotransmission is known to have important arousal/activating functions. The neurons responsible for those actions also release the atypical neuromodulator nitric oxide (NO), which has been shown in previous studies to be involved in the modulation of sleep/wake states. The present investigation, using an animal model (anesthetized cat) tests the hypothesis that NO cooperates with ACh in controlling rhythmic neuronal activity, which may play a role in sleep/wake transition. We have used extracellular singleunit recording of neurons in the dorsal thalamus and visual cortex with simultaneous iontophoretic application of drugs acting upon the NO system: the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine (L-NOArg), NO donors, and 8-bromo-cGMP (which mimics the action of NO). Local inhibition of NOS significantly reduced the activity of recorded cells in both thalamus and visual cortex. The opposite effect was achieved with NO donors application. In cortex, ejection of 8-bromo cGMP or the NO donor diethylamine-nitric oxide (DEA-NO) increased cell firing. Furthermore, the rhythmic firing pattern present in these cortical neurons was disrupted. Taken together, these findings suggest that the NO system collaborates with cholinergic neurotransmission. This collaboration might be involved in the control of different patterns of electrogenic activity during various states of the sleep-wake cycle, via the ability of the NO system to modify rhythmic activity of neurons. PMID- 11007451 TI - Aberrant regulation and function of wild-type p53 in radioresistant melanoma cells. AB - Sporadic human tumors and the hereditary cancer predisposition syndrome Li Fraumeni are frequently associated with mutations in the p53 tumor suppressor gene that compromise its ability to function as a DNA damage checkpoint. A subset of Li-Fraumeni patients with wild-type p53 alleles have mutations in chk2/hcds1, one of the genes signaling the presence of DNA damage to the p53 protein. This suggests that p53 may be kept inactive in human cancer by mutations targeting DNA damage signaling pathways. Melanoma cells are highly radioresistant, yet they express wild-type p53 protein, raising the possibility of defects in the pathways that activate p53 in response to DNA damage. We have described a chk2/hcds1 independent DNA damage signaling pathway that targets Ser-376 within the COOH terminus of p53 for dephosphorylation and leads to increased p53 functional activity. We now report that in several human melanoma cell lines that express wild-type p53, the phosphorylation state of Ser-376 was not regulated by DNA damage. In these cell lines, neither the endogenous wild-type p53 protein nor high levels of ectopic wild-type p53 led to cell cycle arrest or apoptosis. Thus, defective activation of p53 in response to DNA damage may underlie the radioresistance of human melanoma cells. PMID- 11007452 TI - p202 levels are negatively regulated by serum growth factors. AB - p202 is an IFN-inducible phosphoprotein (Mr 52,000) whose expression in transfected cells retards proliferation. Interestingly, the reduced levels of p202 in fibroblasts (in consequence of the expression of antisense to 202 RNA), under reduced serum conditions, increase the susceptibility of cells to apoptosis. To identify the functional role of p202 in cell growth regulation, we tested whether serum growth factor levels in the culture medium affect p202 levels. Here we report that, under reduced serum conditions, the p202 levels were increased in fibroblasts, and the increase was seen at both the mRNA and protein levels. Moreover, an increase in p202 levels was correlated with cell growth arrest in the G1 phase of the cell cycle. Interestingly, the presence of platelet derived growth factor AB, basic fibroblast growth factor, or transforming growth factor beta1 in the culture medium abrogated the increase in p202 levels seen under reduced serum conditions. We found that the increase in p202 levels was accompanied by an increase in JunD/activation protein 1(AP-1) levels, and transfection of a JunD-encoding plasmid along with a reporter plasmid in which transcription of the reporter gene (luciferase) was driven by the 5'-regulatory region of the 202 gene resulted in an increase in the activity of luciferase. Additionally, stable overexpression of JunD in cells, under reduced serum conditions, also resulted in an increase in p202 levels. Interestingly, one of the AP-1-like DNA-binding sequences present in the 5'-regulatory region of the 202 gene could selectively bind to the JunD/AP-1 transcription factor. Taken together, our observations reported herein suggest that in fibroblasts, under reduced serum conditions, the increased levels of JunD/AP-1 contribute to the transcriptional up-regulation of p202 levels, which may be important for the regulation of apoptosis. PMID- 11007453 TI - Inhibition of progesterone-induced Xenopus oocyte maturation by Nm23. AB - The Nm23 protein has been implicated in a wide variety of biological processes, including suppression of metastasis, phytochrome responses in plants, and regulation of differentiation. Here we examine whether Nm23 is involved in Xenopus laevis oocyte maturation. We found that Nm23 is present in oocytes, indicating that it has the potential to be a regulator of maturation. Furthermore, modest overexpression of Nm23 inhibited progesterone-induced oocyte maturation. This maturation-inhibitory activity was shared by both the acidic Nm23-H1 isoform and the basic Nm23-H2 isoform and by Nm23 mutants that lack nucleoside diphosphate kinase activity (Nm23-H1 H118F and Nm23-H2 H118F). Expression of Nm23 proteins delayed the accumulation of Mos and the activation of p42 mitogen-activated protein kinase (MAPK) in progesterone-treated oocytes but had no discernible effect on Mos-induced p42 MAPK activation. Therefore, Nm23 appears to act upstream of the Mos/mitogen-activated protein/extracellular signal regulated kinase kinase/p42 MAPK cascade. These findings suggest a novel biological role for Nm23. PMID- 11007454 TI - Protein kinase C inhibitor and irradiation-induced apoptosis: relevance of the cytochrome c-mediated caspase-9 death pathway. AB - Caspases are a family of cysteine proteases that constitute the apoptotic cell death machinery. We report the importance of the cytochrome c-mediated caspase-9 death pathway for radiosensitization by the protein kinase C (PKC) inhibitors staurosporine (STP) and PKC-412. In our genetically defined tumor cells, treatment with low doses of STP or the conventional PKC-specific inhibitor PKC 412 in combination with irradiation (5 Gy) potently reduced viability, enhanced mitochondrial cytochrome c release into the cytosol, and specifically stimulated the initiator caspase-9. Whereas treatment with each agent alone had a minimal effect, combined treatment resulted in enhanced caspase-3 activation. This was prevented by broad-range and specific caspase-9 inhibitors and absent in caspase 9-deficient cells. The tumor suppressor p53 was required for apoptosis induction by combined treatment but was dispensable for dose-dependent STP-induced caspase activation. These results demonstrate the requirement for an intact caspase-9 pathway for apoptosis-based radiosensitization by PKC inhibitors and show that STP induces apoptosis independent of p53. PMID- 11007455 TI - K562 cells resistant to phorbol 12-myristate 13-acetate-induced growth arrest: dissociation of mitogen-activated protein kinase activation and Egr-1 expression from megakaryocyte differentiation. AB - The K562 cell line undergoes megakaryocytic differentiation in response to phorbol 12-myristate 13-acetate (PMA) stimulation. This event correlates with mitogen-activated protein kinase activation, cell cycle arrest, and expression of the Egr-1 transcription factor. We have isolated K562 cells that are resistant to the growth-inhibitory action of PMA. Molecular characterization demonstrates that PMA resistance is downstream from PMA-induced activation of the mitogen-activated protein kinase pathway. Although the levels of Egr-1 expression and cyclic AMP responsive element-binding protein phosphorylation are comparable in wild-type and PMA-resistant clones in response to PMA, the expression of megakaryocytic cell surface marker CD41 is detected only in the wild-type cells. The lack of differentiation of the PMA-resistant clones correlates with a failure of the PMA treated cells to induce dephosphorylation and down-regulation of the retinoblastoma protein. These cells may provide a useful model system to distinguish those events that are connected to cell cycle arrest from those involved in the differentiation program initiated by PMA. PMID- 11007456 TI - Internal exposure from the ingestion of foods contaminated by 137Cs after the Chernobyl accident--report 2. Ingestion doses of the rural population of Ukraine up to 12 y after the accident (1986-1997). AB - Doses from the ingestion of 134Cs and 137Cs during 12 y following the Chernobyl accident have been estimated for approximately 3 million persons living in rural areas of the Zhitomir, Rivne, and Kyiv Oblasts of northern Ukraine. This assessment is based upon an extensive monitoring campaign that provided measurements of 137Cs in more than 120,000 samples of milk and in more than 100,000 persons; such measurements were made in approximately 4,500 locations. Two approaches were used for the dose assessment. In the first approach a so called reference dose is estimated for each settlement on the basis of measured 137Cs concentration in milk, determination of the milk equivalent of diet, and consumption rates; a further assumption is that a high fraction of the food consumed is produced locally. The reference dose is used as the official dose estimate, which is the basis for any decision on possible financial compensation and economic privileges. In a second step, the so-called real age-dependent dose is estimated from the results of whole body counter measurements and the kinetics of radiocesium in the human body. Real doses above 0.5, 5, and 50 mSv were received by about 40%, 10%, and 0.2%, respectively, of the considered population. With the exception of 1986, for which the monitoring results were limited, the real individual doses derived from whole-body counting are consistently lower than the reference doses. However, this difference declined from a factor of 3-4 in 1987-1989 to a factor of approximately 1.5 in the mid 1990's. The difference between reference and real doses is attributed to the effectiveness of countermeasures implemented after the accident. The effectiveness of these countermeasures decreased with time due to increasing economic problems in Ukraine. The collective reference and real doses of the rural population due to the intake of 134Cs and 137Cs are estimated to be 13,300 and 5,300 person-Sv, respectively. Thus, about 8,000 person-Sv is estimated to have been averted by countermeasures. PMID- 11007457 TI - Accelerator mass spectrometry of 63Ni at the Munich Tandem Laboratory for estimating fast neutron fluences from the Hiroshima atomic bomb. AB - After the release of the present dosimetry system DS86 in 1987, measurements have shown that DS86 may substantially underestimate thermal neutron fluences at large distances (>1,000 m) from the hypocenter in Hiroshima. This discrepancy casts doubts on the DS86 neutron source term and, consequently, the survivors' estimated neutron doses. However, the doses were caused mainly by fast neutrons. To determine retrospectively fast neutron fluences in Hiroshima, the reaction 63Cu(n, p)63Ni can be used, if adequate copper samples can be found. Measuring 63Ni (half life 100 y) in Hiroshima samples requires a very sensitive technique, such as accelerator mass spectrometry (AMS), because of the relatively small amounts of 63Ni expected (approximately 10(5)-10(6) atoms per gram of copper). Experiments performed at Lawrence Livermore National Laboratory have demonstrated in 1996 that AMS can be used to measure 63Ni in Hiroshima copper samples. Subsequently, a collaboration was established with the Technical University of Munich in view of its potential to perform more sensitive measurements of 63Ni than the Livermore facility and in the interest of interlaboratory validation. This paper presents the progress made at the Munich facility in the measurement of 63Ni by AMS. The Munich accelerator mass spectrometry facility is a combination of a high energy tandem accelerator and a detection system featuring a gas-filled magnet. It is designed for high sensitivity measurements of long lived radioisotopes. Optimization of the ion source setup has further improved the sensitivity for 63Ni by reducing the background level of the 63Cu isobar interference by about two orders of magnitude. Current background levels correspond to a ratio of 63Ni/Ni<2x10(-14) and suggest that, with adequate copper samples, the assessment of fast neutron fluences in Hiroshima and Nagasaki is possible for ground distances of up to 1500 m, and--under favorable conditions- even beyond. To demonstrate this capability, we have measured successfully 6Ni/Ni ratios as low as (3.5 +/- 0.6) x 10(-13). The latter are, based on DS86, representative of a ratio expected from a typical Hiroshima copper sample at about 1,300-m ground range. PMID- 11007458 TI - Radon progeny exposure and lung cancer risk among non-smoking uranium miners. AB - Studies of miners provide the basis for public health efforts to reduce residential radon progeny exposure. Because the preponderance of households do not have members who smoke indoors, studies of non-smoking miners contribute essential data for risk assessments for residential radon progeny exposure. We studied a cohort of 2,209 never-smokers who were underground uranium miners employed in the western U.S. from 1956 to the early 1990's and who participated in a screening program for lung cancer conducted by Saccomanno and colleagues. After determining the vital status and cause of death in the cohort, we conducted a nested case-control study of 55 lung cancer deaths in males and 3 age-matched controls for each case. The relative risk of lung cancer was 29.2 (95% CI 5.1, 167.2) for miners with greater than 1,450 WLM compared with those exposed to less than 80 WLM. Temporal factors affected risk, including average dose rate, which was inversely associated with lung cancer risk, and the length of time since last exposure, which was directly associated with decreased risk. As in studies of non smokers and smokers combined, the exposure response relationship in never-smokers was consistent with a decreased slope at higher WLM, which resulted, in part, from an inverse dose rate effect. PMID- 11007459 TI - Restricting exposure to pulsed and broadband magnetic fields. AB - A general procedure is described for application of the new ICNIRP exposure guidelines to pulsed and broadband magnetic fields below 100 kHz. The procedure involves weighting of the spectral components with a function that takes into account the basic restrictions and reference levels. A simple first-order RC response or its piecewise linear equivalent is proposed for the weighting function. The weighting can be performed either on the Fourier transformed sample of the measured signal or in real time by processing the signal with an analog or digital filter circuit. The cut-off frequency of the filter is 820 Hz. The occupational exposure criteria are exceeded when the weighted peak magnetic flux density exceeds 43 microT or equivalently the weighted peak dB/dt exceeds 0.22 T s(-1). The maximal peak exposures allowed by the proposed approach are compared with the stimulation thresholds computed with a stimulation model. The results strongly suggest that the safety margin to the stimulation is greater for non sinusoidal than for sinusoidal waveforms, and at low frequencies it is higher than at high frequencies. The increase of the low-frequency safety margin is desirable to avoid magnetophosphenes and possible CNS effects that may occur below the level predicted by the classical nerve models. Measurement techniques and examples of measured magnetic fields are presented. Particularly high exposures were measured inside MRI equipment and anti-theft gates. PMID- 11007460 TI - Theoretical study of the relation between radon and its long-lived progeny in a room. AB - We present a theoretical study of the complex relation between radon and its long lived progeny implanted in glass surfaces. The well known (extended) Jacobi room model, which is normally used to describe radon and its progeny in a room, was transformed into a two-parameter model revealing a linear correlation between long term radon exposure and surface activity due to implanted radon decay products. Furthermore, this new approach made integration into a Monte Carlo simulation possible so that the large variation of different room model parameters could be taken into account. This allowed the calculation of a probability distribution for radon exposure from the measurement of the implanted 210Po activity. The availability of a 95% confidence interval for the radon exposure is valuable in the application of retrospective radon assessment in epidemiological studies. PMID- 11007461 TI - In situ hot spot measurements with HPGe detector 0.1 M above ground. AB - The objective of this study was to examine the feasibility of an accurate in situ measurement of radioactive hot spots by a HPGe detector located at a height of 0.1 m above the surface, thus sparing the awkward use of a heavy collimator required at the standard 1 m height. Radioactivity concentrations of primordial naturally occurring 40K, 232Th, 238U and of anthropogenic 137Cs in a sand plot were measured in situ at the two heights. Independently, six fractions of a soil profile collected in the site were measured in the laboratory. Good agreement was found between the results of the two geometries and the soil samples. Good agreement between measured and predicted radioactivity concentration values was obtained for hot spots simulated by uniform planar sources of 40K, 60Co and 137Cs having a circular pattern 0.5 m in diameter. The study shows that an in situ measurement without a collimator but close to a hot spot can provide a reliable result. PMID- 11007462 TI - Using measured 30-150 kVp polychromatic tungsten x-ray spectra to determine ion chamber calibration factors, Nx (Gy C(-1)). AB - Two methods for determining ion chamber calibration factors (Nx) are presented for polychromatic tungsten x-ray beams whose spectra differ from beams with known Nx. Both methods take advantage of known x-ray fluence and kerma spectral distributions. In the first method, the x-ray tube potential is unchanged and spectra of differing filtration are measured. A primary standard ion chamber with known Nx for one beam is used to calculate the x-ray fluence spectrum of a second beam. Accurate air energy absorption coefficients are applied to the x-ray fluence spectra of the second beam to calculate actual air kerma and Nx. In the second method, two beams of differing tube potential and filtration with known Nx are used to bracket a beam of unknown Nx. A heuristically derived Nx interpolation scheme based on spectral characteristics of all three beams is described. Both methods are validated. Both methods improve accuracy over the current half value layer Nx estimating technique. PMID- 11007463 TI - On the application of a radiation weighting factor for alpha particles in protection of non-human biota. AB - Radiation protection standards for non-human biota have been expressed in terms of absorbed dose. In calculating dose to biota, some investigators have modified the absorbed dose due to alpha particles by a factor of 20, based on the radiation weighting factor used in protection of humans, to account for the greater effectiveness of these radiations in producing biological damage. However, this value is intended to apply to stochastic health effects, primarily cancers, whereas deterministic effects have been the primary concern in protection of biota. Based on an analysis by the International Commission on Radiological Protection, the deterministic radiation weighting factor for alpha particles appears to lie in the range of about 5-10. Given the potential importance of this weighting factor in determining allowable levels of alpha emitting radionuclides in the environment, regulatory authorities must be impressed with the need to develop an appropriate value for use in protection of biota. There also is a need to express doses and dose limits for biota in terms of a quantity other than absorbed dose and to develop an appropriate name for the biologically significant dose to biota. PMID- 11007464 TI - The LNT-controversy and the concept of "controllable dose". AB - There is no firm scientific information on the potential health effects, such as increased cancer rates, due to low doses of ionizing radiation. In view of this uncertainty ICRP has adopted as a prudent default option the linear no-threshold (LNT) assumption and has used it to derive nominal risk coefficients. Subsequent steps, such as the comparison of putative fatality rates in radiation workers with observed accident rates in other professions, have given the risk estimates a false appearance of scientific fact. This has encouraged meaningless computations of radiation-induced fatalities in large populations and has caused a trend to measure dose limits for the public not against the magnitude of the natural radiation exposure and its geographic variations, but against the numerical risk estimates. In reaction to this development, opposing claims are being made of a threshold in dose for deleterious health effects in humans. In view of the growing polarization, ICRP is now exploring a new concept "controllable dose" that aims to abandon the quantity collective dose, emphasizing, instead, individual dose and, in particular, the control of the maximum individual dose from single sources. Essential features of the new proposal are here examined, and it is concluded that the control of individual dose will still have to be accompanied by the avoidance of unnecessary exposures of large populations, even if their magnitude lies below that acceptable to the individual. If a reasonable cut-off at trivial doses is made, the collective dose can remain useful. Misapplications of collective dose are not the deeper cause of the current controversy; the actual root is the misrepresentation of the LNT assumption as a scientific fact and the amplification of this confusion by loose terminology. If over-interpretation and distortion are avoided, the current system of radiation protection is workable and essentially sound, and there is no need for a fruitless LNT-controversy. The new concept of controllable dose promises simplifications and improvements, but any major change of principles needs to be carefully considered in a broad discussion that ICRP is presently seeking. PMID- 11007465 TI - Internal dosimetry of plutonium using the late urinary excretion. AB - An attempt has been made to standardize the methodology of internal dose computation from the late urinary excretion data. The methodology was selected keeping in mind the most recent ICRP publications and the results of internal dosimetry intercomparison studies reported in literature. The key element of this methodology is the PC-based computational software LUDEP 2.05, which implements the new model of the human respiratory tract. Late urinary excretion data of three male subjects involved in accidental intakes of plutonium aerosols more than 25 years ago were interpreted in terms of intakes and internal doses with the aid of the standardized methodology. An important implication of this work is that late urinary excretion data of the occupational workers of any plutonium handling facility could be used to show the compliance with the life-time dose limit. PMID- 11007466 TI - Self absorption of alpha and beta particles in a fiberglass filter. AB - Environmental air sampling uses fiberglass filters to collect particulate matter from the air and then a gas flow detector to measure the alpha and beta activity on the filter. When counted, the filter is located close to the detector so the alpha and beta particles emerging from the filter travel toward the detector at angles ranging from zero to nearly 90 degrees to the normal to the filter surface. The particles at small angles can readily pass through the filter, but particles at large angles pass through a significant amount of filter material and can be totally absorbed. As a result, counting losses can be great. For 4 MeV alpha particles, the filter used in this experiment absorbs 43% of the alpha particles; for 7.5 MeV alphas, the absorption is 13%. The measured beta activities also can have significant counting losses. Beta particles with maximum energies of 0.2 and 2.0 MeV have absorptions of 44 and 2%, respectively. PMID- 11007467 TI - Revision of guidelines on limits of exposure to laser radiation of wavelengths between 400 nm and 1.4 microm. International Commission on Non-Ionizing Radiation Protection. PMID- 11007468 TI - Nuclear power plant operating experiences: an international report. PMID- 11007470 TI - U.K. mobile phone base station study. PMID- 11007469 TI - NRPB advice on constraints and limits. PMID- 11007471 TI - A novel cytidine deaminase affects antibody diversity. PMID- 11007472 TI - BIRinging chromosomes through cell division--and survivin' the experience. PMID- 11007473 TI - RING finger proteins: mediators of ubiquitin ligase activity. PMID- 11007474 TI - Class switch recombination and hypermutation require activation-induced cytidine deaminase (AID), a potential RNA editing enzyme. AB - Induced overexpression of AID in CH12F3-2 B lymphoma cells augmented class switching from IgM to IgA without cytokine stimulation. AID deficiency caused a complete defect in class switching and showed a hyper-IgM phenotype with enlarged germinal centers containing strongly activated B cells before or after immunization. AID-/- spleen cells stimulated in vitro with LPS and cytokines failed to undergo class switch recombination although they expressed germline transcripts. Immunization of AID-/- chimera with 4-hydroxy-3-nitrophenylacetyl (NP) chicken gamma-globulin induced neither accumulation of mutations in the NP specific variable region gene nor class switching. These results suggest that AID may be involved in regulation or catalysis of the DNA modification step of both class switching and somatic hypermutation. PMID- 11007475 TI - Activation-induced cytidine deaminase (AID) deficiency causes the autosomal recessive form of the Hyper-IgM syndrome (HIGM2). AB - The activation-induced cytidine deaminase (AID) gene, specifically expressed in germinal center B cells in mice, is a member of the cytidine deaminase family. We herein report mutations in the human counterpart of AID in patients with the autosomal recessive form of hyper-IgM syndrome (HIGM2). Three major abnormalities characterize AID deficiency: (1) the absence of immunoglobulin class switch recombination, (2) the lack of immunoglobulin somatic hypermutations, and (3) lymph node hyperplasia caused by the presence of giant germinal centers. The phenotype observed in HIGM2 patients (and in AID-/- mice) demonstrates the absolute requirement for AID in several crucial steps of B cell terminal differentiation necessary for efficient antibody responses. PMID- 11007476 TI - Activation of a membrane-bound transcription factor by regulated ubiquitin/proteasome-dependent processing. AB - Processing of integral membrane proteins in order to liberate active proteins is of exquisite cellular importance. Examples are the processing events that govern sterol regulation, Notch signaling, the unfolded protein response, and APP fragmentation linked to Alzheimer's disease. In these cases, the proteins are thought to be processed by regulated intramembrane proteolysis, involving site specific, membrane-localized proteases. Here we show that two homologous yeast transcription factors SPT23 and MGA2 are made as dormant ER/nuclear membrane localized precursors and become activated by a completely different mechanism that involves ubiquitin/proteasome-dependent processing. SPT23 and MGA2 are relatives of mammalian NF-kappaB and control unsaturated fatty acid levels. Intriguingly, proteasome-dependent processing of SPT23 is regulated by fatty acid pools, suggesting that the precursor itself or interacting partners are sensors of membrane composition or fluidity. PMID- 11007478 TI - Molecular structure of human TFIIH. AB - TFIIH is a multiprotein complex required for both transcription and DNA repair. Single particles of human TFIIH were revealed by electron microscopy and image processing at a resolution of 3.8 nm. TFIIH is 16 x 12.5 x 7.5 nm in size and is organized into a ring-like structure from which a large protein domain protrudes out. A subcomplex assembled from five recombinant core subunits also forms a circular architecture that can be superimposed on the ring found in human TFIIH. Immunolabeling experiments localize several subunits: p44, within the ring structure, forms the base of the protruding protein density which includes the cdk7 kinase, cyclin H, and MAT1. Within the ring structure, p44 was flanked on either side by the XPB and XPD helicases. These observations provide us with a quartenary organizational model of TFIIH. PMID- 11007477 TI - Global role for chromatin remodeling enzymes in mitotic gene expression. AB - Regulation of eukaryotic gene expression requires ATP-dependent chromatin remodeling enzymes, such as SWI/SNF, and histone acetyltransferases, such as Gcn5p. Here we show that SWI/SNF remodeling controls recruitment of Gcn5p HAT activity to many genes in late mitosis and that these chromatin remodeling enzymes play a role in regulating mitotic exit. In contrast, interphase expression of GAL1, HIS3, PHO5, and PHO8 is accompanied by SWI/SNF-independent recruitment of Gcn5p HAT activity. Surprisingly, prearresting cells in late mitosis imposes a requirement for SWI/SNF in recruiting Gcn5p HAT activity to the GAL1 promoter, and GAL1 expression also becomes dependent on both chromatin remodeling enzymes. We propose that SWI/SNF and Gcn5p are globally required for mitotic gene expression due to the condensed state of mitotic chromatin. PMID- 11007479 TI - Electron crystal structure of the transcription factor and DNA repair complex, core TFIIH. AB - Core TFIIH from yeast, made up of five subunits required both for RNA polymerase II transcription and nucleotide excision DNA repair, formed 2D crystals on charged lipid layers. Diffraction from electron micrographs of the crystals in negative stain extended to about 13 angstrom resolution, and 3D reconstruction revealed several discrete densities whose volumes corresponded well with those of individual TFIIH subunits. The structure is based on a ring of three subunits, Tfb1, Tfb2, and Tfb3, to which are appended several functional moieties: Rad3, bridged to Tfb1 by SsI1; SsI2, known to interact with Tfb2; and Kin28, known to interact with Tfb3. PMID- 11007480 TI - Structure of functionally activated small ribosomal subunit at 3.3 angstroms resolution. AB - The small ribosomal subunit performs the decoding of genetic information during translation. The structure of that from Thermus thermophilus shows that the decoding center, which positions mRNA and three tRNAs, is constructed entirely of RNA. The entrance to the mRNA channel will encircle the message when a latch-like contact closes and contributes to processivity and fidelity. Extended RNA helical elements that run longitudinally through the body transmit structural changes, correlating events at the particle's far end with the cycle of mRNA translocation at the decoding region. 96% of the nucleotides were traced and the main fold of all proteins was determined. The latter are either peripheral or appear to serve as linkers. Some may assist the directionality of translocation. PMID- 11007481 TI - Structural basis for relief of autoinhibition of the Dbl homology domain of proto oncogene Vav by tyrosine phosphorylation. AB - Rho-family GTPases transduce signals from receptors leading to changes in cell shape and motility, mitogenesis, and development. Proteins containing the Dbl homology (DH) domain are responsible for activating Rho GTPases by catalyzing the exchange of GDP for GTP. Receptor-initiated stimulation of Dbl protein Vav exchange activity involves tyrosine phosphorylation. We show through structure determination that the mVav1 DH domain is autoinhibited by an N-terminal extension, which lies in the GTPase interaction site. This extension contains the Tyr174 Src-family kinase recognition site, and phosphorylation or truncation of this peptide results in stimulation of GEF activity. NMR spectroscopy data show that the N-terminal peptide is released from the DH domain and becomes unstructured upon phosphorylation. Thus, tyrosine phosphorylation relieves autoinhibition by exposing the GTPase interaction surface of the DH domain, which is obligatory for Vav activation. PMID- 11007482 TI - Src tyrosine kinase is a novel direct effector of G proteins. AB - Heterotrimeric G proteins transduce signals from cell surface receptors to modulate the activity of cellular effectors. Src, the product of the first characterized proto-oncogene and the first identified protein tyrosine kinase, plays a critical role in the signal transduction of G protein-coupled receptors. However, the mechanism of biochemical regulation of Src by G proteins is not known. Here we demonstrate that Galphas and Galphai, but neither Galphaq, Galpha12 nor Gbetay, directly stimulate the kinase activity of downregulated c Src. Galphas and Galphai similarly modulate Hck, another member of Src-family tyrosine kinases. Galphas and Galphai bind to the catalytic domain and change the conformation of Src, leading to increased accessibility of the active site to substrates. These data demonstrate that the Src family tyrosine kinases are direct effectors of G proteins. PMID- 11007483 TI - The serpin alpha1-proteinase inhibitor is a critical substrate for gelatinase B/MMP-9 in vivo. AB - We have identified the key protein substrate of gelatinase B/MMP-9 (GB) that is cleaved in vivo during dermal-epidermal separation triggered by antibodies to the hemidesmosomal protein BP180 (collagen XVII, BPAG2). Mice deficient in either GB or neutrophil elastase (NE) are resistant to blister formation in response to these antibodies in a mouse model of the autoimmune disease bullous pemphigoid. Disease develops upon complementation of GB -/- mice with NE -/- neutrophils or NE -/- mice with GB -/- neutrophils. Only NE degrades BP180 and produces dermal epidermal separation in vivo and in culture. Instead, GB acts upstream to regulates NE activity by inactivating alpha1-proteinase inhibitor (alpha1-PI). Excess NE produces lesions in GB -/- mice without cleaving alpha1-PI. Excess alpha1-PI phenocopies GB and NE deficiency in wild-type mice. PMID- 11007484 TI - The polar T1 interface is linked to conformational changes that open the voltage gated potassium channel. AB - Kv voltage-gated potassium channels share a cytoplasmic assembly domain, T1. Recent mutagenesis of two T1 C-terminal loop residues implicates T1 in channel gating. However, structural alterations of these mutants leave open the question concerning direct involvement of T1 in gating. We find in mammalian Kv1.2 that gating depends critically on residues at complementary T1 surfaces in an unusually polar interface. An isosteric mutation in this interface causes surprisingly little structural alteration while stabilizing the closed channel and increasing the stability of T1 tetramers. Replacing T1 with a tetrameric coiled-coil destabilizes the closed channel. Together, these data suggest that structural changes involving the buried polar T1 surfaces play a key role in the conformational changes leading to channel opening. PMID- 11007485 TI - A novel genetic pathway for sudden cardiac death via defects in the transition between ventricular and conduction system cell lineages. AB - HF-1 b, an SP1 -related transcription factor, is preferentially expressed in the cardiac conduction system and ventricular myocytes in the heart. Mice deficient for HF-1 b survive to term and exhibit normal cardiac structure and function but display sudden cardiac death and a complete penetrance of conduction system defects, including spontaneous ventricular tachycardia and a high incidence of AV block. Continuous electrocardiographic recordings clearly documented cardiac arrhythmogenesis as the cause of death. Single-cell analysis revealed an anatomic substrate for arrhythmogenesis, including a decrease and mislocalization of connexins and a marked increase in action potential heterogeneity. Two independent markers reveal defects in the formation of ventricular Purkinje fibers. These studies identify a novel genetic pathway for sudden cardiac death via defects in the transition between ventricular and conduction system cell lineages. PMID- 11007486 TI - A FRET-based sensor reveals large ATP hydrolysis-induced conformational changes and three distinct states of the molecular motor myosin. AB - The molecular motor myosin is proposed to bind to actin and swing its light-chain binding region through a large angle to produce an approximately 10 nm step in motion coupled to changes in the nucleotide state at the active site. To date, however, direct dynamic measurements have largely failed to show changes of that magnitude. Here, we use a cysteine engineering approach to create a high resolution, FRET-based sensor that reports a large, approximately 70 degree nucleotide-dependent angle change of the light-chain binding region. The combination of steady-state and time-resolved fluorescence resonance energy transfer measurements unexpectedly reveals two distinct prestroke states. The measurements also show that bound Mg.ADP.Pi, and not bound Mg.ATP, induces the myosin to adopt the prestroke states. PMID- 11007488 TI - Why is there late replication? AB - It has been known for about 40 years that the S phase of the cell cycle is regulated and that parts of the genome are replicated early, while others are replicated late. Numerous studies in the past two decades have revealed that while expressed genes, such as those coding for housekeeping proteins, are usually replicated early, genes not expressed in a particular cell and heterochromatic regions of the genome, such as the centromeres or the inactivated X chromosome of females, are usually replicated late. As details of the mechanisms leading to the formation of replication complexes were worked out, in particular for the budding yeast, Saccharomyces cerevisiae, new insights into the control of the order of replication of genes were obtained that indicate that this process is highly regulated. It is coordinated with transcription, epigenetic changes in chromatin structure, regulation of precursor pools and surveillance mechanisms. PMID- 11007487 TI - CLIP170-like tip1p spatially organizes microtubular dynamics in fission yeast. AB - Rod-shaped fission yeast cells grow in a polarized manner, and unlike budding yeast, the correct positioning of the growth sites at cell ends requires interphase microtubules. Here we describe a microtubule guidance mechanism that orients microtubules in the intracellular space along the long axis of the cell, guiding them to their target region at the cell ends. This mechanism involves tip1p, a CLIP170-like protein that localizes to distal tips of cytoplasmic microtubules. In the absence of tip1p, microtubular catastrophe is no longer restricted to cell ends but occurs when microtubules reach any region of the cellular cortex. Thus, tip1p enables microtubules to discriminate different cortical regions and regulates their dynamics accordingly. PMID- 11007489 TI - The chromosomal distribution of phosphorylated histone H3 differs between plants and animals at meiosis. AB - Plant (Secale cereale, Triticum aestivum) and animal (Eyprepocnemis plorans) meiocytes were analyzed by indirect immunostaining with an antibody recognizing histone H3 phosphorylated at serine 10, to study the relationship between H3 phosphorylation and chromosome condensation at meiosis. To investigate whether the dynamics of histone H3 phosphorylation differs between chromosomes with a different mode of segregation, we included in this study mitotic cells and also meiotic cells of individuals forming bivalents plus three different types of univalents (A chromosomes, B chromosomes and X chromosome). During the first meiotic division, the H3 phosphorylation of the entire chromosomes initiates at the transition from leptotene to zygotene in rye and wheat, whereas in E. plorans it does so at diplotene. In all species analyzed H3 phosphorylation terminates toward interkinesis. The immunosignals at first meiotic division are identical in bivalents and univalents of A and B chromosomes, irrespective of their equational or reductional segregation at anaphase I. The grasshopper X chromosome, which always segregates reductionally, also shows the same pattern. Remarkable differences were found at second meiotic division between plant and animal material. In E. plorans H3 phosphorylation occurred all along the chromosomes, whereas in plants only the pericentromeric regions showed strong immunosignals from prophase II until telophase II. In addition, no immunolabeling was detectable on single chromatids resulting from equational segregation of plant A or B chromosome univalents during the preceding anaphase I. Simultaneous immunostaining with anti-tubulin and anti-phosphorylated H3 antibodies demonstrated that the kinetochores of all chromosomes interact with microtubules, even in the absence of detectable phosphorylated H3 immunosignals. The different pattern of H3 phosphorylation in plant and animal meiocytes suggests that this evolutionarily conserved post-translational chromatin modification might be involved in different roles in both types of organisms. The possibility that in plants H3 phosphorylation is related to sister chromatid cohesion is discussed. PMID- 11007490 TI - A neocentromere in the DAZ region of the human Y chromosome. AB - We describe a novel rearranged human Y chromosome consisting of an inverted duplication of the long arm heterochromatin and a small amount of euchromatin: rea(Y)(qter-q11.2::q11.2-qter). The normal centromere has been deleted and a neocentromere containing CENP-A, -C, -E and Mad2 but not CENP-B has formed close to the breakpoint. A 2.7 Mb yeast artificial chromosome contig spanning the breakpoint was constructed and the breakpoint was localised to a region of <120 kb close to the DAZ gene cluster. Combined immunofluorescence and fluorescence in situ hybridisation showed that the centromeric protein-binding domain of the neocentromere was located near the breakpoint and within the DAZ cluster. PMID- 11007491 TI - CENP-G in neocentromeres and inactive centromeres. AB - CENP-G is a novel constitutive centromere-specific protein localized to the kinetochore inner plate and subjacent region. It has been identified as associating specifically with the alpha-1 subfamily of alpha-satellite DNA. In the present work, the localization of CENP-G was compared with that of other CENPs by immunofluorescence and fluorescence in situ hybridization. Studies were carried out on four abnormal human centromeres: two neocentromeres and two inactive centromeres. CENP-G was detected in one of the two inactive centromeres but not in the other that shows a partial deletion of the alphoid DNA. Interestingly, CENP-G is also present in neocentromeres, which lack alphoid DNA sequences, and in the human Y chromosome, which lacks the alpha-1 type of satellite DNA. These data provide further evidence that CENP-G may be an essential factor in centromeric function and that in centromeres lacking the alpha-1 subfamily of alphoid DNA, other DNA sequences are able to bind CENP-G. PMID- 11007492 TI - A modifier screen of ectopic Kruppel activity identifies autosomal Drosophila chromosomal sites and genes required for normal eye development. AB - Irregular facets (If) is a dominant gain-of-function allele of the Drosophila segmentation gene Kruppel (Kr) that interferes with eye development. In a search for genes that interact with Kr activity, we recently performed a systematic genetic screen to identify dominant enhancers and suppressors of the If eye phenotype that are located on the third chromosome. Here we describe locations and candidate genes of the second chromosome that act as dominant modifiers of ectopic Kr activity during eye development. The collection of more than 40 modifiers of Kr activity located on the second and third chromosomes, from which a total of 16 genes were identified, includes genes encoding transcription factors and components of signal transduction pathways that may regulate or be regulated by Kr activity. We also identified genes coding for more general cellular factors that could interfere with the intracellular transport or the half-life of the Kr protein. The data demonstrate that the If mutation provides a means to screen the Drosophila genome for functional components of developmental pathways that depend on or can be modified by Kr activity. Owing to the bias of the screening system applied, these modifier genes will be expressed and are likely to be required during Drosophila wild-type eye development. PMID- 11007493 TI - Structural evolution of the germ line-limited chromosomes in Acricotopus. AB - The elimination of chromatin or whole chromosomes from the future somatic nuclei during germ line-soma differentiation in early embryogenesis is a genetic phenomenon found in a wide variety of animal species. Less is known about the origin, structure, and function of the germ line-limited chromosomes. In the chironomid Acricotopus lucidus fluorescence in situ hybridization (FISH) with labeled soma DNA to "Keimbahn" chromosomes (Ks) and soma chromosomes (Ss) of spermatogonial mitoses revealed that each of the nine different K types possesses large S-homologous sections, mostly in the distal parts of both chromosome arms. Painting probes of the three Ss and of each of their chromosome arms were generated by microdissection of polytene salivary gland chromosomes and subsequent amplification by the degenerate oligonucleotide-primed polymerase chain reaction. Multicolor FISH demonstrated that each of the Ks, with the exception of one K type, was painted by only one of the three S probes. Furthermore, in seven Ks, one chromosome arm was painted by the long-arm probe and the other by the short-arm probe of the S concerned. The hybridization pattern strongly suggests that each of these K types is derived from a specific S. One function of the S-homologous K sections is thought to be determination of the regular occurrence of crossover events, with the resulting chiasmata in these sections ensuring correct segregation of the K homologs during meiosis. Reverse chromosome painting on polytene S sets with a probe generated from metaphase Ks corroborates the above results and produces conclusive evidence for the hypothesis that during evolution the Ks have developed from the Ss by endopolyploidization and rearrangements followed by the accumulation of germ line specific repetitive DNA sequences in the centromeric regions. PMID- 11007494 TI - Invasion of Drosophila virilis by the Penelope transposable element. AB - The Penelope family of transposable elements (TEs) is broadly distributed in most species of the virilis species group of Drosophila. This element plays a pivotal role in hybrid dysgenesis in Drosophila virilis, in which at least four additional TE families are also activated. Here we present evidence that the Penelope family of elements has recently invaded D. virilis. This evidence includes: (1) a patchy geographical distribution, (2) genomic locations mainly restricted to euchromatic chromosome arms in various geographical strains, and (3) a high level of nucleotide similarity among members of the family. Two samples from a Tashkent (Middle Asia) population of D. virilis provide further support for the invasion hypothesis. The 1968 Tashkent strain is free of Penelope sequences, but all individuals collected from a 1997 population carry at least five Penelope copies. Furthermore, a second TE, Ulysses, has amplified and spread in this population. These results provide evidence for the Penelope invasion of a D. virilis natural population and the mobilization of unrelated resident transposons following the invasion. PMID- 11007495 TI - Genetic induction of chromosomal rearrangements in barley chromosome 7H added to common wheat. AB - Chromosome 2C of Aegilops cylindrica induces chromosomal rearrangements in alien chromosome addition lines, as well as in euploid lines, of common wheat. To induce chromosomal rearrangements in barley chromosome 7H, reciprocal crosses were made between a mutation-inducing common wheat line that carries a pair of 7H chromosomes and one 2C chromosome and a 7H disomic addition line of common wheat. Many shrivelled seeds were included in the progeny, which was an indication of the occurrence of chromosome mutations. The chromosomal constitution of the viable progeny was examined by FISH (fluorescence in situ hybridization) using the barley subterminal repeat HvT01 as a probe. Structural changes of chromosome 7H were found in about 15% of the progeny of the reciprocal crosses. The aberrant 7H chromosomes were characterized by a combination of N-banding, FISH and genomic in situ hybridization. Mosaicism for aberrant 7H chromosomes was observed in seven plants. In total, 89 aberrant 7H chromosomes were identified in 82 plants, seven of which had double aberrations. More than half of the plants carried a simple deletion: four short-arm telosomes, one long-arm telosome, and 45 terminal deletions (23 in the short arm, 21 in the long arm, and one involving both arms). About 40% of the aberrations represented translocations between 7H and wheat chromosomes. Twenty of the translocations had wheat centromeres, 12 the 7H centromere, with translocation points in the 7HS (five) and in the 7HL (seven), and the remaining four were of Robertsonian type, three involving 7HS and one with 7HL. In addition, one translocation had a barley segment in an intercalary position of a wheat chromosome, and two were dicentric. The breakpoints of these aberrations were distributed along the entire length of chromosome 7H. PMID- 11007496 TI - Prof. Anthony J. Raimondi. PMID- 11007497 TI - History of the Korean Neurosurgical Society and future prospects: how should we educate trainee pediatric neurosurgeons? PMID- 11007498 TI - Reprint of "Concerning Surgical Intervention for the Intracranial Hemorrhages of the New-born" by Harvey Cushing, M.D. 1905. AB - This paper was first presented at a meeting in 1905 and gives an insight into the problems faced in early neurosurgical operations in newborns and the way they were approached by an adventurous surgeon. PMID- 11007499 TI - Sub-galeal coiling of the proximal and distal components of a ventriculo peritoneal shunt. An unusual complication and proposed mechanism. AB - We describe the case of a child in whom proximal migration of the peritoneal catheter and extrusion of the ventricular catheter resulted in the entire ventriculo-peritoneal shunt along with the shunt chamber (Orbis Sigma II valve) lying in a sub-galeal pocket in the occipital region in a tightly coiled fashion. This coiling was very similar in appearance to that of the pre-insertion shunt in the packaging when it is supplied; hence it is postulated that the migration was secondary to retained 'memory' of the shunt tubing. This is a rare complication of ventriculo-peritoneal shunt, which has not been described before. PMID- 11007500 TI - Basilar artery occlusion in a child: "clot angioplasty" followed by thrombolysis. AB - Basilar artery occlusions are rare but have a very poor prognosis. Intra-arterial thrombolysis may produce recanalization and better clinical outcome. A short delay between the onset of symptoms and thrombolysis is considered essential for successful recanalization and for the smallest possible risk of haemorrhagic complications. We present a case of basilar artery occlusion in an 8-year-old child, which was treated by "clot angioplasty" followed by intra-arterial thrombolysis. Thirty hours after progressive alteration of consciousness, speech disturbances and left arm paresis, the child became comatose with decerebrate rigidity. A CT scan showed parenchymal ischaemic lesions. Angiography (performed 36 h after the onset of symptoms) showed a total occlusion of the basilar artery. A clot angioplasty was performed by placing a balloon catheter within the thrombus and inflating it several times in the occluded segment of the basilar artery. Thrombolysis was then performed through the balloon catheter. The basilar artery was only partially recanalized at the end of the procedure, but the perforating arteries of the brain stem had reappeared on angiography. Three months later the child had completely recovered to a normal clinical status. In conclusion, the very poor natural prognosis of basilar artery occlusion requires aggressive management. Recanalization of the basilar artery may be performed even late after the onset of symptoms. Clot angioplasty allows partial recanalization, which may increase the efficiency of thrombolysis. PMID- 11007501 TI - Congenital brain tumor in a neonate conceived by in vitro fertilization. AB - Congenital brain tumors are very rare; their incidence is estimated at 0.34 per million live births. We report a case of congenital gliosarcoma in a neonate conceived by in vitro fertilization (IVF). One other case of brain tumor (medulloblastoma) is reported in a child born after assisted conception. Whether these tumors are causally related to the IVF remains obscure. PMID- 11007502 TI - Craniocerebral injury resulting from transorbital stick penetration in children. AB - OBJECTS: Two children were admitted to hospital for treatment of craniocerebral injury with transorbital penetration. METHODS: One child aged 6 years and 6 months had poked a chopstick in his orbit. There was no report of either a palpebral or an ocular wound. He had subsequently developed a meningeal syndrome with a cerebral abscess managed by needle aspiration biopsy and intravenous antibiotics. The other child, aged 4, had fallen onto a metal rod. He presented with a palpebral wound, motor disorders and coma, all due to a frontal intracerebral hematoma. There was an improvement in outcome without complications of an infectious nature or motor sequelae. CONCLUSIONS: Such head injuries are rare. Clinical, radiological and ophthalmological investigations must be performed, including computed tomography (CT) scan or cerebral magnetic resonance imaging (MRI) with antibiotic treatment for suspected microorganisms. PMID- 11007503 TI - Spinal type IV arteriovenous malformations (perimedullary fistulas) in children. AB - OBJECTS: Intradural perimedullary spinal arteriovenous fistula (AVF) is a synonym for type IV spinal arteriovenous malformation (AVM). It is an important clinical differential diagnosis in all patients with slowly progressive or acute spinal symptoms. Perimedullary AVFs are rare in the paediatric age group. We report the treatment regimen and the clinico-radiological findings for these malformations at our institution. METHODS: Of four paediatric patients, three individuals with persistent fistulas after endovascular treatment were operated on. The malformations were obliterated completely in all patients without any morbidity. In three patients previously documented neurological symptoms resolved during follow-up. CONCLUSIONS: If slowly progressive or acute radicular or medullary symptoms arise in children, a spinal arteriovenous malformation should be ruled out by MRI. A combined endovascular and surgical treatment of paediatric spinal AVM type IV (perimedullary AVFs) carries a low risk of morbidity and is reasonably effective. PMID- 11007504 TI - Aberrant expression of neurotrophic factors in the ventricular progenitor cells of infant congenitally hydrocephalic rats. AB - OBJECTS: This study was conducted to investigate the roles of neurotrophic factors in the development of hydrocephalus in HTX rats. METHODS: Expressions of brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and fibroblast growth factor (FGF)-1 were examined immunohistochemically in the cerebral cortex and ventricular zone of 6-day-old rats with congenital hydrocephalus (HTX rats). In the ventricular zone of hydrocephalic rats, potent BDNF-like immunoreactivity (-LI) and weak but significant signals for NT-3- and FGF-1-LIs were observed. However, no significant signals were detected in non-HTX rats. A small subpopulation of ventricular cells was positive for microtubule-associated protein 2 in HTX and non-HTX rats. The positive cells in the HTX rats had neurites much longer than those in the non-HTX animals, suggesting that some ventricular cells of the hydrocephalics had ectopically differentiated into mature neurons. CONCLUSIONS: This abnormal differentiation may have been responsible for the aberrant expressions of neurotrophic factors. In contrast, the cerebral neuronal layers did not show such prominent alterations in neurotrophic factor expression. PMID- 11007505 TI - Diagnostic steps and staged operative approach in Currarino's triad: a case report and review of the literature. AB - The Currarino triad is a combination of a presacral mass, a congenital sacral bony abnormality and an anorectal malformation. It mostly presents with constipation. Rectal examination, plain radiographs and magnetic resonance imaging are the main tools for the diagnosis. If the mass is a meningocele, colostomy and neurosurgical exploration should precede anoplasty due to the risk of meningitis. A 14-month-old female patient with anal stenosis, a sacral scimitar defect and an anterior meningocele is presented in this report. PMID- 11007506 TI - Nuclear medicine in the next decade. EANM Working Party. PMID- 11007507 TI - Combination of clinical and V/Q scan assessment for the diagnosis of pulmonary embolism: a 2-year outcome prospective study. AB - With the aim of evaluating the efficiency of our diagnostic approach in patients with suspected acute pulmonary embolism (PE), we prospectively studied 143 patients investigated by means of a ventilation/perfusion (V/Q) lung scan. A pre test clinical probability of PE (Pclin) was assigned to all patients by the clinicians and scans were interpreted blinded to clinical assessment. A 2-year follow-up of our patients was systematically performed and possible in 134 cases. Distribution of clinical probabilities was high Pclin in 22.5%, intermediate Pclin in 24% and low Pclin in 53.5%, whereas the distribution of scan categories was high Pscan in 14%, intermediate Pscan in 18%, low Pscan in 57% and normal Pscan in 11%. The final prevalence of PE was 24.5%. High Pscan and normal Pscan were always conclusive (19 and 15 cases respectively). Low Pscan associated with low Pclin could exclude PE in 43/45 cases (96%). None of the patients in whom the diagnosis of PE was discarded had a major event related to PE during the 2-year follow-up. Overall, the combined assessment of clinical and scintigraphic probabilities allowed confirmation or exclusion of PE in 80% of subjects (107/134) and proved to be a valuable tool for selecting patients who needed pulmonary angiography, which was required in 20% of our patients (27/134). PMID- 11007508 TI - Radioiodine therapy for Plummer's disease based on the thyroid uptake of technetium-99m pertechnetate. AB - The aim of this retrospective study was the evaluation of a TcTUs (global technetium-99m pertechnetate thyroid uptake under suppression)-based approach in 370 patients with thyroid autonomy (Plummer's disease) treated by radioiodine therapy (RIT) under standardised conditions. The analysis included 370 patients (309 females, 61 males; mean age 64+/-11.6 years) treated for thyroid autonomy [unifocal (UFA), 36.8%; multifocal (MFA), 55.7%; disseminated (DISA), 7.6%]. During RIT all patients were under thyroid suppression (TSH< 0.1 microU/ml) and without thionamide treatment. Of the 370 patients, 73% (n=271) were manifestly hyperthyroid and 27% (n=99) subclinically hyperthyroid. A dosimetric study included uptake measurements 24, 48, and 96 h p.i. For dose estimation the Marinelli algorithm was used. For retrospective definition of the target volume we used the equation: Autonomous volume = TcTUs x 5. The spectrum of doses given in our patients ranged from 81 to 1933 Gy. After 18 months of follow-up, RIT was successful (TSH>0.5 microqU/l and/or TcTUs<1.6%) in 310 patients (84%). Of these patients, 291 (94%) were euthyroid (with or without L-thyroxine) and 19 (6%) subclinically hypothyroid (TSH>4 microU/ml). A dose of 350-450 Gy to the autonomous tissue resulted in a success rate of 97% in the UFA group and 81% in the MFA/DISA group. Decrease in total thyroid volume and TcTUs did not differ significantly between successfully treated patients and patients with persistent autonomy. Multivariate analysis of all 370 patients identified four independent factors that negatively influenced the therapeutic success: high pretherapeutic thyroid volume (P=0.0001; odds ratio: 1.017), high pretherapeutic TcTUs values (P=0.0001; odds ratio: 1.378), multifocal/disseminated autonomy (P=0.0056; odds ratio: 3.245) and low target dose (P=0.017; odds ratio: 0.997). It is concluded that the high success rate in the treatment of UFA indicates the concept of TcTUs based RIT to be valid, but that in the therapy of MFA/DISA the target dose has to be corrected if the total thyroid volume exceeds a critical threshold. PMID- 11007509 TI - Changes in myocardial perfusion after catheter-based percutaneous laser revascularisation. AB - This study investigated the effect of percutaneous laser revascularisation (PMR) on regional myocardial blood flow. PMR is a new therapeutic modality for patients suffering from angina pectoris due to coronary artery disease (CAD) that is not amenable to revascularisation. Initial clinical studies discovered reduced angina pectoris and increased exercise capacity after PMR. There are no previous clinical studies reporting the impact of PMR on perfusion. Thirty-six patients with end-stage CAD underwent thallium-201 single-photon emission tomography studies on four different occasions: immediately before PMR and 3, 6 and 12 months following PMR. Each study consisted of pharmacological stress and rest scintigraphy. Semiquantitative evaluation was performed by use of 14 standardised wall segments which were classified (a) according to the localisation of the PMR target region into treated segments, segments adjacent to the treated area and non-treated segments and (b) according to the pre-therapeutic local perfusion into four groups: no (0), slight (I), moderate (II) or severe (III) perfusion deficit. At stress in treated segments of groups II and III local scintigraphic count densities increased from 60.1% and 34.7% at baseline to 65.3% and 48.3% after 12 months (P<0.05) while they decreased in segments of group 0 from 94.2% to 85.7% (P<0.05). In rest studies no changes occurred. Thus, the local rest stress differences within the target areas become smaller after PMR. In the PMR target area but not in the nontreated area an improvement in regional myocardial flow reserve occurs in wall segments with initially severely or moderately reduced stress perfusion. This effect is consistent with the clinical improvement after PMR. PMID- 11007510 TI - 99mTc-MIBI radio-guided minimally invasive parathyroid surgery planned on the basis of a preoperative combined 99mTc-pertechnetate/99mTc-MIBI and ultrasound imaging protocol. AB - The aims of this study were: (a) to define the accuracy of a preoperative parathyroid imaging protocol based on the combination of technetium-99m pertechnetate/technetium-99m methoxyisobutylisonitrile (99mTcO4/ 99mTc-MIBI) scan and neck ultrasound (US) in selecting patients with primary hyperparathyroidism (pHPT) eligible for a limited neck exploration, and (b) to investigate the potential role of the intraoperative gamma probe (IGP) in radio-guided minimally invasive surgery. 99mTcO4/99mTc-MIBI subtraction scan was performed by means of potassium perchlorate administration with the aim of effecting rapid 99mTcO4 wash out from the thyroid. Minimally invasive surgery using an IGP was commenced some minutes following the injection of a low, 70 MBq, 99mTc-MIBI dose. Intraoperative PTH (i-PTH) was measured. On the basis of preoperative imaging, 21 pHPT consecutive patients were selected for a limited neck dissection. In 18 of them, a single parathyroid adenoma was found at surgery and IGP allowed performance of parathyroidectomy through a small, 2-2.5 cm, skin incision with a relatively short surgical duration (mean 38 min). i-PTH rapidly normalised in all cases. In two patients, a parathyroid carcinoma was diagnosed at surgery; consequently, a wide neck exploration associated with a near-total thyroidectomy was performed. No loco-regional metastatic lesions were found and i-PTH rapidly normalised after carcinoma excision. In one patient, i-PTH remained elevated after removal of the enlarged parathyroid gland which was localised by 99mTcO4/99mTc-MIBI scan and US. A bilateral exploration was needed to remove a contralateral enlarged parathyroid gland. Combined, 99mTcO4/99mTc-MIBI scan and US imaging correctly localised a single parathyroid gland in 20/21 patients (95.2%); thus, this protocol appears to be accurate enough for the preoperative selection of pHPT patients eligible for limited neck surgery. Moreover, in these selected patients the IGP seems to be helpful in performing radio-guided minimally invasive surgery. PMID- 11007511 TI - FDG-PET for detection of osseous metastases from malignant primary bone tumours: comparison with bone scintigraphy. AB - The purpose of this study was to compare positron emission tomography using fluorine-18 fluorodeoxyglucose (FDG-PET) and technetium-99m methylene diphosphonate (MDP) bone scintigraphy in the detection of osseous metastases from malignant primary osseous tumours. In 70 patients with histologically proven malignant primary bone tumours (32 osteosarcomas, 38 Ewing's sarcomas), 118 FDG PET examinations were evaluated. FDG-PET scans were analysed with regard to osseous metastases in comparison with bone scintigraphy. The reference methods for both imaging modalities were histopathological analysis, morphological imaging [additional conventional radiography, computed tomography (CT) or magnetic resonance imaging (MRI)] and/or clinical follow-up over 6-64 months (median 20 months). In 21 examinations (18%) reference methods revealed 54 osseous metastases (49 from Ewing's sarcomas, five from osteosarcomas). FDG-PET had a sensitivity of 0.90, a specificity of 0.96 and an accuracy of 0.95 on an examination-based analysis. Comparable values for bone scintigraphy were 0.71, 0.92 and 0.88. On a lesion-based analysis the sensitivity of FDG-PET and bone scintigraphy was 0.80 and 0.72, respectively. Analysing only Ewing's sarcoma patients, the sensitivity, specificity and accuracy of FDG-PET and bone scan were 1.00, 0.96 and 0.97 and 0.68, 0.87 and 0.82, respectively (examination-based analysis). None of the five osseous metastases from osteosarcoma were detected by FDG-PET, but all of them were true-positive using bone scintigraphy. In conclusion, the sensitivity, specificity and accuracy of FDG-PET in the detection of osseous metastases from Ewing's sarcomas are superior to those of bone scintigraphy. However, in the detection of osseous metastases from osteosarcoma, FDG-PET seems to be less sensitive than bone scintigraphy. PMID- 11007512 TI - Cholecystokinin receptor imaging using an octapeptide DTPA-CCK analogue in patients with medullary thyroid carcinoma. AB - Cholecystokinin (CCK)-B receptors have been demonstrated on a high percentage of medullary thyroid carcinomas (MTC) in vitro. After encouraging results both in vitro and in animal studies, we studied the efficacy of an octapeptide [111In DTPA]-CCK analogue in seven patients with MTC. In four of five patients in whom serum calcitonin levels were monitored, a significant rise was found following the injection, indicating retained biological activity of the radiopeptide. In all patients there was visualization of the CCK-B receptor-positive stomach. In one of two patients with known MTC lesions, some of the lesions were visualized; in addition some lesions were visualized in one of the five other patients who had elevated serum tumour markers but negative localizing studies. Radioactivity in the presumed tumour sites was still present at 48 h p.i. The uptake in the presumed tumour sites and stomach was low. Background radioactivity dropped rapidly owing to urinary excretion. After 1 h, breakdown products of the labelled analogue predominated both in urine and in serum, and virtually no intact peptide was present. IN CONCLUSION: (1) the CCK-B receptor-positive gastric mucosa and presumed MTC lesions could be visualized in patients using an octapeptide [111In DTPA]-CCK analogue that is probably internalized, proving the feasibility of CCK B receptor imaging in vivo; (2) there was a relatively low uptake of the CCK analogue in the strongly CCK receptor positive stomach, and rapid degradation of the peptide in serum. PMID- 11007513 TI - 99mTc-EDDA/HYNIC-TOC: a new 99mTc-labelled radiopharmaceutical for imaging somatostatin receptor-positive tumours; first clinical results and intra-patient comparison with 111In-labelled octreotide derivatives. AB - [111In-diethylene triamine penta-acetic acid-D-Phe1]-octreotide (DTPA-octreotide) scintigraphy has gained widespread acceptance as a diagnostic clinical procedure in oncology for imaging somatostatin receptor-positive tumours. However, indium 111 as a radiolabel has several drawbacks, including limited availability, suboptimal gamma energy and high radiation burden to the patient. We have recently reported on the preclinical development of 99mTc-EDDA/HYNIC-TOC, a new octreotide derivative which showed promising results both in vitro and in vivo. We now report our initial clinical experiences with this new radiopharmaceutical in ten oncological patients. The clinical diagnoses were: carcinoid syndrome (n=5), thyroid cancer (n=3), pancreatic cancer (n=1) and pituitary tumour (n=1). The biodistribution and kinetics of 99mTc-EDDA/HYNIC-TOC were compared with those of 111In-DTPA-octreotide in six cases, and with those of 111In-DOTA-TOC in five cases. With the new tracer tumours were imaged within 15 min after injection and showed the highest target/non-target ratios 4 h after injection. Tumour uptake persisted up to 20 h p.i. The rate of blood clearance was similar to that of 111In-DTPA-octreotide but faster than that of 111In-DOTA-TOC, while urinary excretion was lower compared with the 111In derivatives. Semi-quantitative region of interest analysis showed that 99mTc-EDDA/HYNIC-TOC produced higher tumour/organ (target/non-target) ratios than the 111In derivatives, especially in relation to heart and muscle. Significantly more lesions could be detected in 99mTc images. We conclude that 99mTcEDDA/HYNIC-TOC shows better imaging properties for the identification of somatostatin receptor-positive tumour sites than currently available 111In-labelled octreotide derivatives. PMID- 11007514 TI - Biodistribution and dosimetry of [123I]iodo-PK 11195: a potential agent for SPET imaging of the peripheral benzodiazepine receptor. AB - The highest concentrations of the peripheral benzodiazepine receptor (PBR) are found in the kidneys and heart. In addition, the PBR has been reported to reflect neuro-inflammatory damage by co-localisation with activated microglia. PK 11195 is a high-affinity ligand for the PBR. The aim of this study was to investigate in humans the biodistribution and dosimetry of [123I]iodoPK 11195, a potential single-photon emission tomography tracer for the PBR. Five healthy volunteers were injected with 112 MBq of [123I]iodo-PK 11195. Sequential whole-body scans were performed up to 72 h post injection. Multiple blood samples were taken, and urine was collected to measure the fraction voided by the renal system. Decay corrected regions of interest of the whole-body images were analysed, and geometric mean count rates were used to determine organ activity. Organ absorbed doses and effective dose were calculated using the MIRD method. [123I]iodo-PK 11195 was rapidly cleared from the blood, mainly by the hepatobiliary system. Approximately 22% was voided in urine after 48 h. Average organ residence times were 0.74, 0.44 and 0.29 h for the liver, upper large intestine and lower large intestine, respectively. The testes received the highest dose, 109.4 microGy/MBq. All other organs investigated received doses of less than 50 microGy/MBq. The effective dose was 40.3 microSv/MBq. In conclusion, [123I]iodo-PK 11195 is a suitable agent for the visualisation of the PBR and indirectly for the imaging of neuro-inflammatory lesions. Taking into account the radiation burden of 7.46 mSv following an administration of 185 MBq, a [123I]iodo-PK 11195 investigation has to be considered an ICRP risk category IIb investigation. PMID- 11007515 TI - Monoclonal antibody-dendrimer conjugates enable radiolabeling of antibody with markedly high specific activity with minimal loss of immunoreactivity. AB - For the purpose of radioimmunotherapy, labelling of monoclonal antibody with high specific activity is often necessary, especially when using a radionuclide with a shorter half-life. Polyamine dendrimers (PAMAM) are novel synthetic polymeric molecules with large numbers of amine residues on their spherical surface. In order to bind large numbers of radiometals to single antibody molecules, the generation-4 PAMAM (G4), which has 64 amines, was conjugated with 43 molecules of 2-(p-isothiocyanatobenzyl)-6-methyl-diethylene triamine penta-acetic acid (1B4M), a derivative of DTPA. This product [G4-(1B4M)43] was then conjugated with OST7, a murine monoclonal IgG1. We evaluated the achievable specific activity for 111In labeling, immunoreactivity, biodistribution, and tumor targeting in mice of the 111In- or 153Gd-OST7-G4-(1B4M)43 as compared with radiolabeled OST7-1B4M or 56C 1B4M. The maximum specific activity of 111In-OST7-G4-(1B4M)43 and 111In-OST7-1B4M was 470 and 8.7 GBq/mg (12,700 and 263 mCi/mg), respectively. Immunoreactivity of radiolabeled OST7-G4-(1B4M)43 and OST7-1B4M, as determined by the binding to KT005 cells expressing the antigen, was respectively 91% and 84% of that of 125I labelled OST7. Biodistribution studies for preparations with maximum specific activity in normal mice 3 h after injection showed that 111In- or 153Gd-OST7-G4 (1B4M)43 cleared faster from the blood and accumulated more in the liver than did 111In- or 153Gd-OST7-1B4M. The dendrimer 1B4M [G4-(1B4M)64] itself showed similar saturation effects with metals. The radioactivity in all the other organs reflected the rapid clearance of radioactivity from the blood. 153Gd-OST7-G4 (1B4M)43 showed specific accumulation in the KT005 tumor. In conclusion, we could successfully bind 49 times as many metal atoms to an antibody molecule as is possible with conventional metal labeling for indium and gadolinium, and did so with minimal loss of immunoreactivity. When we achieved radiolabeling with maximum specific activity, Gd conjugate showed better biodistribution than In conjugate. PMID- 11007517 TI - Influence of arm positioning on tomographic thallium-201 myocardial perfusion imaging and the effect of attenuation correction. AB - Lateral attenuation in single-photon emission tomography (SPET) myocardial perfusion imaging (MPI) has been attributed to the left arm if it is held by the patient's side during data acquisition. As a result MPI data are conventionally acquired with the arms held above the head. The aims of this study were to determine the effect of imaging arms down on reconstructed tomographic images depicting regional myocardial thallium-201 distribution and to assess whether attenuation-corrected (AC) myocardial perfusion images acquired arms down could replace uncorrected (NC) images acquired arms up for routine clinical service. Twenty-eight patients referred for routine MPI underwent sequential 180 degrees emission/transmission imaging for attenuation correction using an L-shaped dual headed gamma camera (GE Optima) fitted with two gadolinium-153 scanning line sources. Delay data were acquired twice: once supine with the arms up and then supine with the arms down. Detector radius of rotation (ROR) for arms up and arms down studies was recorded. For each data set, count density was measured in 17 segments of a polar plot and segmental uptake expressed relative to study maximum. Oblique images were assessed qualitatively by two observers blinded to study type for tracer distribution and overall quality. Transmission maps were assessed for truncation. Mean detector ROR was 190 mm for arms-up studies and 232 mm for arms-down studies (P<0.05). Population mean segmental relative uptake values for NC arms-up studies were higher than for NC arms-down studies, with the greatest difference seen anterolaterally. Nevertheless, the majority (24/28) of oblique NC arms-up and NC arms-down images appeared similar and only four (14%) NC arms-down studies showed additional areas of reduced count density (one anterior and three lateral). Corresponding AC arms-down studies showed that count density within the anterior defect improved to normal but the lateral reductions persisted, and in two of these three studies the arms-down transmission map was distorted. Population mean segmental relative uptake values for NC arms-down studies were lower than for AC arms-down studies apart from three anterolateral segments where NC arms-down values were higher. Of 28 AC arms-down studies, 11 (39%) were of reduced quality compared with NC arms-up studies because of poorer spatial resolution and because AC enhances liver activity compared with NC. It is concluded that arm positioning influences reconstructed tomographic images depicting regional 201T1 distribution, particularly anterolaterally. There is lateral undercorrection in approximately 10% of AC arms-down studies, possibly because of attenuation map truncation. Image quality is reduced in about one third of AC armsdown studies compared with NC arms-up studies. These data suggest that this attenuation correction method is not sufficiently robust to allow routine acquisition of MPI data with the arms down. PMID- 11007516 TI - Significant regional heterogeneity of coronary flow reserve in paediatric hypertrophic cardiomyopathy. AB - Previous studies have indicated that cardiac events in young patients with hypertrophic cardiomyopathy (HCM) are related to ischaemia rather than to arrhythmia. We measured coronary flow reserve in paediatric HCM and compared the values with those in adult HCM. We studied 12 patients with HCM including six paediatric (<20 years old; mean 13 years) and six adult patients (>20 years old: mean 62 years), and six healthy young adults (mean 29 years) as controls. Every patient underwent magnetic resonance imaging (MRI) for anatomical assessment. Myocardial blood flow at rest and after dipyridamole infusion was measured with dynamic nitrogen-13 ammonia positron emission tomography (PET). Partial volume effect was corrected for using the anatomical data obtained with MRI. In adult patients with HCM, coronary flow reserve in the hypertrophied septal region was not significantly different from that in the non-hypertrophied lateral wall (1.38+/-0.29 vs 1.77+/-0.39, respectively). In the paediatric patients, coronary flow reserve in the hypertrophied septal region was significantly lower than in the non-hypertrophied lateral wall (0.84+/-0.33 vs 2.74+/-0.90, respectively, P<0.01). In addition, coronary flow reserve in adult patients was lower than in control subjects both in the septal wall (1.38+/-0.29 vs 2.94+/-0.35, respectively, P<0.0001) and in the lateral wall (1.77+/-0.39 vs 2.85+/-0.69, respectively, P<0.05). In contrast, coronary flow reserve in paediatric patients was not significantly different from that in control subjects in the lateral wall (2.74+/-0.90 vs 2.85+/-0.69, respectively), while absolute reduction of myocardial blood flow was noted after pharmacological vasodilatation in the hypertrophied septal region. In conclusion, significant regional differences of coronary flow reserve were present in the paediatric patients with HCM. These results suggest that paediatric patients with HCM intrinsically have the potential to experience significant regional ischaemia even in the absence of coronary stenosis. PMID- 11007518 TI - Should scatter be corrected in both transmission and emission data for accurate quantitation in cardiac SPET? AB - Ideally, reliable quantitation in single-photon emission tomography (SPET) requires both emission and transmission data to be scatter free. Although scatter in emission data has been extensively studied, it is not well known how scatter in transmission data affects relative and absolute quantitation in reconstructed images. We studied SPET quantitative accuracy for different amounts of scatter in emission and transmission data using a Utah phantom and a cardiac Data Spectrum phantom including different attenuating media. Acquisitions over 180 degrees were considered and three projection sets were derived: 20% images and Jaszczak and triple-energy-window scatter-corrected projections. Transmission data were acquired using gadolinium-153 line sources in a 90-110 keV window using a narrow or wide scanning window. The transmission scans were performed either simultaneously with the emission acquisition or 24 h later. Transmission maps were reconstructed using filtered backprojection and mu values were linearly scaled from 100 to 140 keV. Attenuation-corrected images were reconstructed using a conjugate gradient minimal residual algorithm. The mu value underestimation varied between 4% with a narrow transmission window in soft tissue and 22% with a wide window in a material simulating bone. Scatter in the emission and transmission data had little effect on the uniformity of activity distribution in the left ventricle wall and in a uniformly hot compartment of the Utah phantom. Correcting the transmission data for scatter had no impact on contrast between a hot and a cold region or on signal-to-noise ratio (SNR) in regions with uniform activity distribution, while correcting the emission data for scatter improved contrast and reduced SNR. For absolute quantitation, the most accurate results (bias <4% in both phantoms) were obtained when reducing scatter in both emission and transmission data. In conclusion, trying to obtain the same amount of scatter in emission and transmission data, in addition to being impractical because of the difficulty in knowing the precise scatter components, did not yield such accurate absolute activity quantitation as when emission and transmission scatter were reduced. PMID- 11007519 TI - Impact of attenuation correction on the accuracy of FDG-PET in patients with abdominal tumors: a free-response ROC analysis. AB - The aim of this study was to evaluate image quality and lesion detectability with and without attenuation correction in patients with abdominal tumors, using a free-response receiver operating characteristic (FROC) methodology. Thirty-four patients with various abdominal tumors were evaluated (11 men, 23 women, median age 48 years). Whole-body emission scans were performed 68 min (35-102 min) after intravenous injection of 4.3 MBq/kg fluorine-18 fluorodeoxyglucose (FDG). Images were reconstructed using the OS-EM algorithm and corrected for attenuation either using postinjection singles transmission (n=27) or by calculation and body outline (n=7). Total scan duration did not exceed 70 min. Studies were read independently by four observers unaware of any clinical data. The uncorrected (UC) images were systematically read before the attenuation-corrected (AC) images. All studies were given an image quality score ranging from 1 (unreadable) to 5 (excellent). Each focus of increased activity was then localized and given a probability of malignancy using a five-point scale. The average image quality score was similar for both UC and AC images. At the time of the positron emission tomography (PET) scans, 127 lesions (63 liver metastases, 9 retroperitoneal lesions, 50 peritoneal or bowel lesions, and 5 pancreatic carcinomas) were revealed by pathological or correlative studies. The areas under the FROC curves were consistently greater for AC images (range 0.8663-0.8867) than for UC images (range 0.7774 -0.8613). Overall, the difference between the AC images and the UC images was significant (P=0.019). In particular, correction for attenuation increased the sensitivity regardless of the location of the lesions. In conclusion, correction for attenuation significantly improves the diagnostic accuracy of FDG-PET for abdominal staging of neoplasms, without impairing the image quality. PMID- 11007520 TI - Gated SPET quantification of small hearts: mathematical simulation and clinical application. AB - Quantification of gated single-photon emission tomography (SPET) in small hearts has been considered to be inaccurate. To evaluate the validity of gated SPET in a small chamber volume, mathematical simulation and clinical application to paediatric patients were performed. Myocardium with various chamber sizes from 14 ml to 326 ml was generated assuming an arbitrary resolution (6.9-15.7 mm in full width at half-maximum), noise and zooming factors. The cut-off frequency of the Butterworth filter for preprocessing was varied from 0.16 to 0.63 cycles/cm. The chamber volume was calculated by quantitative gated SPET software (QGS). The patients, aged 2 months to 19 years (n=27), were studied by gated technetium-99m methoxyisobutylisonitrile or tetrofosmin SPET. Image magnification as large as possible was performed during data acquisition to include the whole chest using 1.25-2.0 zooming. Based on the simulation study, an underestimation of the chamber volume occurred below a volume of 100 ml. The degree of underestimation for a 37-ml volume was 49% without zooming, but it improved to 3% with 2x zooming. Filters with a higher cut-off frequency, better system resolution and hardware zooming during acquisition improved quantitative accuracy in small hearts. For the subjects under 7 years old (n=7), quantification of volume and ejection fraction (EF) was possible in 72% of the patients. In those over 7 years old, gated SPET quantification was feasible in all cases. The correlation between gated SPET end-diastolic volume (SPET EDV) and both echocardiographic end diastolic dimension (EDD) and echocardiographic EDV was good (r=0.84 between SPET EDV and echo EDD, r=0.85 between SPET EDV and echo EDV, P<0.0001 for both). The correlation between gated SPET EF and both echocardiographic fractional shortening (FS) and echocardiographic EF was fair (r=0.69 between SPET EF and echo FS, r=0.72 between SPET EF and echo EF, P<0.0001 for both). In conclusion, quantification of gated SPET of small hearts can be improved by means of a SPET filter with a high cut-off frequency, high system resolution and appropriate zooming. Gated SPET should be attempted not only in patients with small hearts but also in paediatric patients. PMID- 11007521 TI - Improvement of brain perfusion SPET using iterative reconstruction with scatter and non-uniform attenuation correction. AB - Filtered back-projection (FBP) is generally used as the reconstruction method for single-photon emission tomography although it produces noisy images with apparent streak artefacts. It is possible to improve the image quality by using an algorithm with iterative correction steps. The iterative reconstruction technique also has an additional benefit in that computation of attenuation correction can be included in the process. A commonly used iterative method, maximum-likelihood expectation maximisation (ML-EM), can be accelerated using ordered subsets (OS EM). We have applied to the OS-EM algorithm a Bayesian one-step late correction method utilising median root prior (MRP). Methodological comparison was performed by means of measurements obtained with a brain perfusion phantom and using patient data. The aim of this work was to quantitate the accuracy of iterative reconstruction with scatter and non-uniform attenuation corrections and post filtering in SPET brain perfusion imaging. SPET imaging was performed using a triple-head gamma camera with fan-beam collimators. Transmission and emission scans were acquired simultaneously. The brain phantom used was a high-resolution three-dimensional anthropomorphic JB003 phantom. Patient studies were performed in ten chronic pain syndrome patients. The images were reconstructed using conventional FBP and iterative OS-EM and MRP techniques including scatter and nonuniform attenuation corrections. Iterative reconstructions were individually post-filtered. The quantitative results obtained with the brain perfusion phantom were compared with the known actual contrast ratios. The calculated difference from the true values was largest with the FBP method; iteratively reconstructed images proved closer to the reality. Similar findings were obtained in the patient studies. The plain OS-EM method improved the contrast whereas in the case of the MRP technique the improvement in contrast was not so evident with post filtering. PMID- 11007522 TI - Comparison of internal radiation doses estimated by MIRD and voxel techniques for a "family" of phantoms. AB - The aim of this study was to use a new system of realistic voxel phantoms, based on computed tomography scanning of humans, to assess its ability to specify the internal dosimetry of selected human examples in comparison with the well established MIRD system of mathematical anthropomorphic phantoms. Differences in specific absorbed fractions between the two systems were inferred by using organ dose estimates as the end point for comparison. A "family" of voxel phantoms, comprising an 8-week-old baby, a 7-year-old child and a 38-year-old adult, was used and a close match to these was made by interpolating between organ doses estimated for pairs of the series of six MIRD phantoms. Using both systems, doses were calculated for up to 22 organs for four radiopharmaceuticals with widely differing biodistribution and emission characteristics (technetium-99m pertechnetate, administered without thyroid blocking; iodine-123 iodide; indium 111 antimyosin; oxygen-15 water). Organ dose estimates under the MIRD system were derived using the software MIRDOSE 3, which incorporates specific absorbed fraction (SAF) values for the MIRD phantom series. The voxel system uses software based on the same dose calculation formula in conjunction with SAF values determined by Monte Carlo analysis at the GSF of the three voxel phantoms. Effective doses were also compared. Substantial differences in organ weights were observed between the two systems, 18% differing by more than a factor of 2. Out of a total of 238 organ dose comparisons, 5% differed by more than a factor of 2 between the systems; these included some doses to walls of the GI tract, a significant result in relation to their high tissue weighting factors. Some of the largest differences in dose were associated with organs of lower significance in terms of radiosensitivity (e.g. thymus). In this small series, voxel organ doses tended to exceed MIRD values, on average, and a 10% difference was significant when all 238 organ doses were considered as a single group. In 12 comparisons of effective dose, the mean voxel to MIRD ratio was 1.07 (range 0.72 1.32). It was shown for the majority of cases that, whereas some large differences in SAF values exist, differences in source organ and effective dose values between the MIRD and voxel methods were largely accounted for by the respective organ mass differences. The reasons for various organ dose differences with the selected radiopharmaceuticals are discussed. Taking biological variation into account, there is reasonable agreement between the two methods but some significant differences exist that warrant further investigation. More extensive comparisons involving a wide variety of voxel phantoms are required to establish whether realistic voxel phantoms should eventually replace the MIRD system. PMID- 11007523 TI - Detecting the sentinel lymph node in patients with differentiated thyroid carcinoma. AB - The aim of this study was to evaluate sentinel lymph node mapping in patients with differentiated thyroid carcinoma. Nine patients with suspected thyroid carcinoma who were scheduled to undergo thyroidectomy underwent scintigraphic localization of sentinel lymph nodes (SLNs). On the day of surgery we injected 37 MBq technetium-99m nanocolloid intratumourally. Dynamic data up to 10 min followed by planar anterior and lateral oblique images up to 1 h after tracer administration were recorded. At surgery the primary tumour was excised first, then the SLNs were removed using a gamma probe. Four patients had papillary carcinoma, two follicular carcinoma, one an oncocytic tumour and two benign tumours. An SLN was identified in all four patients with papillary carcinoma. In the two patients with follicular carcinoma, SLN detection failed. Five patients had one radioactive node, one had three and one had four. In one patient, no SLN was visible with scintigraphic imaging but at surgery three SLNs could be clearly identified using the gamma probe after removal of the primary tumour. There were no false-negative findings. This initial study indicates that in patients with papillary thyroid carcinoma detection of the SLN is possible, whereas the technique failed in two patients with follicular carcinoma. A study on a larger patient sample is now warranted. PMID- 11007524 TI - Technetium-99m DTPA inhalation scintigraphy in patients treated with fluoxetine and maprotiline: preliminary results. AB - Drug-metabolising enzymatic activities have been detected in tracheobronchiolar, bronchiolar and alveolar regions in the lungs. Induction of phospholipidosis by amine drugs such as clorphentermine has also been shown. This study aimed to investigate the effect of fluoxetine and maprotiline, which contain amine groups in their structure, on pulmonary epithelial membrane permeability. Twenty-seven patients (mean age 36+/-12 years) with various psychiatric problems, of whom 17 were treated with fluoxetine and 10 with maprotiline, were included in this study. Technetium-99m diethylene triamine penta-acetic acid (DTPA) aerosol inhalation scintigraphy was performed before and after 4-6 weeks of therapy. Following the inhalation of 1480 MBq 99mTc-DTPA for 3 min, lung images in a 64x64 matrix were obtained every minute for 30 min. Regions of interest were drawn around the periphery of the lungs and on the major airways. Clearance half-times (T 1/2) were calculated by placing a mono-exponential fit on the curves. Penetration index (PI) was calculated on the first-minute image. There was no difference between the clearance rates of 99mTc-DTPA before and after therapy for either the fluoxetine or the maprotiline group. After therapy, a significant decrease in PI was found in patients treated with fluoxetine (PI values before and after therapy: 0.53+/-0.03 and 0.49+/-0.05 respectively, P< or =0.05). This finding might have been due to the induction of increased synaptic serotonin (5 HT) by fluoxetine, which acts by inhibiting the re-uptake of 5-HT on presynaptic membranes. Bronchoconstriction of small and medium airways may be caused by direct and indirect effects of 5-HT on smooth muscle contraction. PMID- 11007525 TI - 99mTc(V)DMSA quantitatively predicts 188Re(V)DMSA distribution in patients with prostate cancer metastatic to bone. AB - Rhenium-188 dimercaptosuccinic acid complex [188Re(V)DMSA], a potential therapeutic analogue of the tumour imaging agent 99mTc(V)DMSA, is selectively taken up in bone metastases in patients with prostate cancer. It would be helpful in planning palliative radionuclide therapy if 99mTc(V)DMSA could be used to predict tumour and kidney retention of 188Re(V)DMSA. The aim of this study was to determine the correlation between tumour-to-normal tissue ratios and kidney-to soft tissue ratios of 99mTc(V)DMSA and 188Re(V)DMSA. This would determine whether a scan with 99mTc(V) DMSA could be used to identify patients for whom 188Re(V)DMSA treatment would be contra-indicated, and enable prediction of relative kidney and tumour radiation absorbed dose in 188Re(V)DMSA treatment. Ten patients with prostate carcinoma were recruited following observation of disseminated bone metastases on a recent 99mTc-hydroxydiphosphonate bone scan. Whole-body planar scans were obtained at ca. 4 h and 24 h after hydration and injection of 600 MBq 99mTc(V)DMSA, and a week later, at similar times after hydration and injection of 370 MBq 188Re(V)DMSA. A triple-energy window (TEW) scatter correction was applied to the 188Re scans. Counts per pixel were determined in regions of interest drawn over metastatic sites, kidneys and normal soft tissue. Tumour-to-soft tissue ratios were significantly lower (by a factor of approximately 0.8 after the TEW was applied) on 188Re scans than on 99mTc scans, but the two were highly linearly correlated both in all individual patients and in tumours pooled from all patients together both at 4 h and at 24 h. Kidney-to-soft tissue ratios were similarly correlated and were lower for 188Re than for 99mTc by a similar factor. Both tumour- and kidney-to-soft tissue ratios increased between 4 and 24 h but the latter increased more. In conclusion, only minor differences were seen between 99mTc and 188Re scans, and kidney-to background ratios on 188Re scans were not higher than on 99mTc scans. These differences are insufficient to infer that they are due to a real difference in biodistribution, and they may be due only to different physical imaging characteristics. Thus 99mTc(V)DMSA scans are predictive of 188Re(V)DMSA biodistribution and could be used to estimate tumour and renal dosimetry and assess suitability of patients for 188Re(V)DMSA treatment. PMID- 11007526 TI - Impaired dopaminergic neurotransmission in patients with traumatic brain injury: a SPECT study using 123I-beta-CIT and 123I-IBZM. AB - Structural imaging suggests that traumatic brain injury (TBI) may be associated with disruption of neuronal networks, including the nigrostriatal dopaminergic pathway. However, to date deficits in pre- and/or postsynaptic dopaminergic neurotransmission have not been demonstrated in TBI using functional imaging. We therefore assessed dopaminergic function in ten TBI patients using [123I]2-beta carbomethoxy-3-beta-(4-iodophenyl)tropane (beta-CIT) and [123I]iodobenzamide (IBZM) single-photon emission tomography (SPET). Average Glasgow Coma Scale score (+/-SD) at the time of head trauma was 5.8+/-4.2. SPET was performed on average 141 days (SD +/-92) after TBI. The SPET images were compared with structural images using cranial computerised tomography (CCT) and magnetic resonance imaging (MRI). SPET was performed with an ADAC Vertex dual-head camera. The activity ratios of striatal to cerebellar uptake were used as a semiquantitative parameter of striatal dopamine transporter (DAT) and D2 receptor (D2R) binding. Compared with age-matched controls, patients with TBI had significantly lower striatal/cerebellar beta-CIT and IBZM binding ratios (P< or =0.01). Overall, the DAT deficit was more marked than the D2R loss. CCT and MRI studies revealed varying cortical and subcortical lesions, with the frontal lobe being most frequently affected whereas the striatum appeared structurally normal in all but one patient. Our findings suggest that nigrostriatal dysfunction may be detected using SPET following TBI despite relative structural preservation of the striatum. Further investigations of possible clinical correlates and efficacy of dopaminergic therapy in patients with TBI seem justified. PMID- 11007527 TI - Experience with carbon-11 choline positron emission tomography in prostate carcinoma. AB - We investigated the potential of carbon-11 choline positron emission tomography (PET) for the detection of lymph node and bone metastases in prostate cancer. A total of 23 patients were studied (known metastases: 8; suspicion of metastases: 3; primary staging: 12). Whole-body PET imaging was performed 5 min after injection of the tracer and completed within 1 h. Focally increased tracer uptake in bone or abdominal lymph node regions was interpreted as representing tumour involvement. All known bone and lymph node metastases could be recognized by [11C]choline PET. One out of ten negative scans for primary staging was false negative (lymph node <1 cm) and one out of two positive scans was false-positive with regard to lymph node involvement (focal bowel activity). It is concluded that [11C]choline PET is a promising new tool for the primary staging of prostate cancer, with lymph node and bone metastases demonstrating high tracer uptake. Therapeutic management could be influenced by these results in that the technique may permit avoidance of surgical lymph node exploration. PMID- 11007528 TI - The importance of the control group in functional brain imaging. PMID- 11007529 TI - Radiolabeled estradiol derivatives to predict response to hormonal treatment in breast cancer: a review. AB - Several radiolabeled steroidal and nonsteroidal estradiol derivatives of which the tumoral uptake is believed to relate quantitatively to the content and binding characteristics of the alpha-estrogen receptor (alphaER) receptor in the target tissue have been synthesized and their imaging potential and clinical usefulness evaluated in vivo in humans. Due to the use of different methodologies and cut-off values for the measurement of alphaER positivity, the use of both quantitative positron emission tomography and semiquantitative single-photon emission tomography, and the difference in patient populations studied, direct comparison of these data is not possible. Individual data, however, fail to substantiate a direct relationship between these radiolabeled estradiol derivatives and alphaER status, in keeping with recent pathophysiological findings demonstrating (1) estradiol sequestration and retention through other than alphaER-mediated, either membrane- or non-membrane-related, mechanisms and (2) an inverse relationship between estradiol uptake and local biosynthesis through aromatization and interconversion in alphaER-positive tumors. Additionally, given the discovery of very high affinity alphaER-like binding sites (Kd, dissociation constant, <0.1 nM), and the potential for underestimation of aER Kd when using ligand binding assays, at least part of the radiolabeled estradiol derivative uptake reflects tumoral perfusion rather than the ligand receptor binding process. However, the reduction in cellular uptake, membrane sequestration and local biosynthesis of estradiol following tamoxifen treatment in alphaER-responsive tumors should allow early prediction of response to therapy through rapid sequential radiolabeled estradiol scintigraphy with higher accuracy than conventional alphaER estimations, as supported by recent data. PMID- 11007531 TI - Effects of soybean lipoxygenase on Na+/K+-ATPase activity in vitro. AB - Oxidized metabolites of polyunsaturated fatty acids produced by lipoxygenase are among the endogenous regulators of Na+/K+-ATPase. The direct effect of lipoxygenase on Na+/K+-ATPase activity was assessed in vitro using soybean lipoxygenase. Treatment of 4.2 microg/mL Na+/K+-ATPase (from dog kidneys) with 4.2 microg/mL of soybean lipoxygenase caused 20+/-2% inhibition of ATPase activity. A 10-fold increase in lipoxygenase concentration (41.6 microg/mL) led to 30+/-0.3% inhibition. In the presence of 12 microg/mL phenidone (a lipoxygenase inhibitor) and 15.4 microg/mL glutathione (a tripeptide containing a cysteine residue) inhibition of Na+/K+-ATPase activity was blocked and an increase in ATPase activity was observed. The presence of lipoxygenase enhanced the inhibition of Na+/K+-ATPase activity caused by 20 ng/mL ouabain (31+/-2 vs. 19+/-2) but had little or no effect with higher concentrations of ouabain. These findings suggest that lipoxygenase may regulate Na+/K+-ATPase by acting directly on the enzyme. PMID- 11007530 TI - The negative inotropic action of catecholamines: role of beta3-adrenoceptors. AB - There is now evidence for the involvement of four beta-adrenoceptor populations in the regulation of cardiac function by catecholamines. Beta1- and beta2 adrenoceptor stimulation classically produces an increase in contractility. A fourth beta-adrenoceptor, as yet uncloned and designated provisionally as a beta4 adrenoceptor, also mediates a positive inotropic effect. Beta3-adrenoceptors, which had been cloned at the end of the eighties, has been extensively studied as a potential target for antiobesity and antidiabetic drugs. Its characterization in the heart has opened new fields of investigations for the understanding of the cardiac adrenergic regulation. This review describes the cardiac electrical and mechanical effects induced by Beta3-adrenoceptor stimulation in different species (including human), as well as the signaling pathway. It also analyzes the role of these receptors in the abnormal responsiveness of catecholamines in heart failure. PMID- 11007532 TI - Tetraethylammonium-evoked oscillatory contractions of rat tail artery: a K-K model. AB - Spontaneously rhythmic contraction of peripheral blood vessels actively modulates the peripheral circulation and blood pressure. However, the underlying mechanisms for the complex rhythmic contraction patterns of various vascular tissues are not yet fully understood. In the present study, the tetraethylammonium (TEA)-induced spontaneously oscillatory contractions of isolated rat tail artery tissues were examined. It was found that TEA evoked arterial oscillatory contractions in a concentration-dependent, but endothelium-independent manner. The voltage dependent K+ (Kv) channel specific blocker, 4-aminopyridine (4-AP), induced a sustained, but not oscillated, vascular contraction. The presence of 4-AP had no effect on the TEA-induced oscillatory contractions. The blockade of KCa channels with charybdotoxin or apamin did not affect the basal force of vascular tissues. Neither the TEA-induced oscillatory contraction was affected by these blockers. The opening of KATP channels by levcromakalim or their blockade by glybenclamide ceased or increased, respectively, the oscillation of TEA-induced contractions. The absence of Ca2+ or the presence of nifedipine in the bath solution completely abolished the effects of TEA. The inhibition of Ca2+-ATPase in the sarcoplasmic reticulum with micromolar concentrations of thapsigargin or cyclopiazonic acid either abolished or enhanced, respectively, the TEA-induced oscillatory contractions. Ryanodine did not affect the TEA-induced oscillatory contraction. In conclusion, the TEA-induced oscillatory contraction may be initiated by the blockade of the TEA-sensitive delayed rectifier K+ channels and maintained by the TEA-insensitive but ATP-sensitive K+ channels. This K-K model presents a novel mechanism for the depolarization-induced rhythmic contractions of small arteries. PMID- 11007533 TI - Selective potentiating effect of RS14203 on a serotoninergic pathway in anesthetized rats. AB - The usefulness of selective inhibitors of type 4 phosphodiesterase (PDE4) in the treatment of inflammation and pulmonary diseases is limited by their side effects: nausea and vomiting. We studied the effect of three structurally diverse PDE4 inhibitors on the vagal nerve afferent and efferent fibers in anesthetized rats. The effects of RS14203, (R)-rolipram, and CT-2450 were evaluated on the von Bezold-Jarisch reflex (vagal afferent fibers) and in a model of vagal electrical stimulation (vagal efferent fibers). All three PDE4 inhibitors were administered at 1, 10, or 100 microg/kg (iv) 15 min prior to the induction of bradycardia by an iv injection of 2-methyl-5-HT (von Bezold-Jarisch reflex) or by vagal electrical stimulation. At 100 microg/kg, RS14203 significantly potentiated the 2 methyl-5-HT response. No statistically significant effects were observed with (R) rolipram or CT-2450 at the doses studied. RS14203, (R)-rolipram, or CT-2450 (1 100 microg/kg iv) did not affect the bradycardia induced by vagal electrical stimulation. Consequently, our results show that RS14203 selectively facilitates serotoninergic neurotransmission in vagal afferent fibers. The emetic action of RS14203 may be mediated by this mechanism. PMID- 11007534 TI - Influence of T-type Ca2+ (mibefradil) and Cl- (indanyloxyacetic acid 94) channel antagonists on alpha1-adrenoceptor mediated contractions in rat aorta. AB - The effects of the T-type and L-type Ca2+ channel antagonists, mibefradil and nifedipine, respectively, and those of a Cl- channel antagonist, indanyloxyacetic acid 94, on mechanical responses elicited by selective activation of alpha1 adrenoceptors using cirazoline were examined in rat isolated aortic rings. The presence of mibefradil (300 nM), indanyloxyacetic acid, 94 (30 microM) and nifedipine (300 nM) alone inhibited mechanical responses elicited by cirazoline. The concentration-response curves to cirazoline were displaced to the right with significant increases in the EC50 and significant depressions of the maximal responses in the presence of the individual agents mibefradil, indanyloxyacetic acid 94, or nifedipine. A combination of mibefradil and indanyloxyacetic acid 94 further inhibited the mechanical activity produced by cirazoline. The further reduction in the maximal response to cirazoline, in the presence of mibefradil and nifedipine, was insignificant when compared with the effects of nifedipine alone. In addition, maximal mechanical responses produced by cirazoline were not significantly affected by a combination of nifedipine and indanyloxyacetic acid 94 when compared with either nifedipine alone or mibefradil and indanyloxyacetic acid 94 combined. Our current findings indicate that mibefradil, indanyloxyacetic acid 94, and nifedipine can inhibit cirazoline-induced contractions to a varying degree. Moreover, based on our present data it would be reasonable to suggest that the contribution of T-type versus L-type Ca2+ channels to contractile responses obtained with cirazoline are approximately 21% and 35%, respectively, of the Emax. It would appear that L-type Ca2+ channels play a greater role in processes that are involved in excitation-contraction coupling subsequent to stimulation of alpha1-adrenoceptors. In addition, Cl- channels also appear to be involved in the process of contraction following alpha1-adrenoceptor activation. PMID- 11007535 TI - Radiotelemetric monitoring of blood pressure and mesenteric arterial bed responsiveness in rats with streptozotocin-induced diabetes. AB - We investigated the changes in arterial blood pressure (BP) and of mesenteric arterial bed (MAB) responsiveness that accompany streptozotocin (STZ)-induced diabetes. BP was recorded by radiotelemetry in conscious animals before and during a 4-week period following induction of the diabetic state with STZ. At the end of this period, the MAB was isolated and perfused under constant flow conditions: perfusion pressure (PP, mmHg) was taken as an index of arteriolar tone. BP was lower (P < 0.05) in STZ-treated diabetic rats (82.9+/-5.0 mmHg) than in vehicle-treated rats (108.9+/-6.3 mmHg). Basal perfusion pressure of the MAB was lower in STZ-treated rats than in control rats and inhibition of nitric oxide (NO) synthesis with N(G)-nitro-L-arginine-methyl-ester and N(G)-nitro-L-arginine (100 microM each) failed to change this relationship. Increases in PP of MAB to phenylephrine (Phe), norepinephrine (NE), and potassium chloride (KCl) were reduced in STZ-treated rats compared with control rats. Inhibition of NO synthesis reduced responses to Phe, NE, and KCL in both STZ and control rats. The reduced responsiveness of STZ rats to Phe, NE, and KCl persisted after inhibition of NO synthesis. Acetylcholine (ACh) evoked relaxation of the MAB in a dose dependent fashion. Maximal responses to ACh, but not sodium nitroprusside, were lower in STZ rats than in vehicle treated rats. Inhibition of NO synthesis reduced responses to ACh in both STZ and control rats. The reduced responsiveness of STZ rats to ACh persisted after inhibition of NO synthesis. The data demonstrate that STZ-induced diabetes is associated with a fall in blood pressure when pressure is recorded with radiotelemetry. The fall in blood pressure may be related to a non-specific decrease in responsiveness to vasoconstrictor stimuli mediated at least in part by NO-independent mechanisms. A decrease in responsiveness to endothelial dependent vasodilator mechanisms appeared insufficient to restore responsiveness to vasoconstrictor stimuli. PMID- 11007536 TI - Activation of Rho signaling contributes to lysophosphatidic acid-induced contraction of intact ileal smooth muscle of guinea-pig. AB - To elucidate the possible role of Rho A/Rho-kinase on lysophosphatidic acid (LPA) induced contraction in intact guinea-pig ileal smooth muscle, we examined effects of pretreatment with a specific inhibitor of Rho-kinase (Y-27632) on the LPA induced contraction and MLC20 phosphorylation. In addition, we investigated whether LPA actually elicits an activation of Rho A by studying subcellular distribution of Rho A in unstimulated and stimulated smooth muscles by LPA. LPA induced a less intense, but sustained, contraction compared with ACh, and was accompanied by significant increases in MLC20 phosphorylation. The effects of LPA on tension and MLC20 phosphorylation were inhibited by Y-27632. The ACh-induced contraction, but not increases in MLC20 phosphorylation, was partially inhibited by Y-27632. High K+-induced contraction was unaffected by the inhibitor. LPA stimulated translocation of Rho A from the cytosol to the membrane fraction of the muscle. Translocation of Rho A was also induced by ACh and high K+. These results suggest that LPA-induced contraction of intact ileal smooth muscle is dominated through activation of Rho A and Rho-kinase and subsequent increases in MLC20 phosphorylation. PMID- 11007537 TI - Effect of stevioside on PAH transport by isolated perfused rabbit renal proximal tubule. AB - Stevioside, a non-caloric sweetening agent, is used as a sugar substitute. An influence of stevioside on renal function has been suggested, but little is known about its effect on tubular function. Therefore, the present study was designed to explore the direct effect of stevioside on transepithelial transport of p aminohippurate (PAH) in isolated S2 segments of rabbit proximal renal tubules using in vitro microperfusion. Addition of stevioside at a concentration of 0.45 mM to either the tubular lumen, bathing medium, or both at the same time had no effect on transepithelial transport of PAH. Similarly, a concentration of 0.70 mM (maximum solubility in the buffer) when present in the lumen, had no effect on PAH transport. However, this concentration in the bathing medium inhibited PAH transport significantly by about 25-35%. The inhibitory effect of stevioside was gradually abolished after it was removed from the bath. Addition of 0.70 mM stevioside to both lumen and bathing medium at the same time produced no added inhibitory effect. Stevioside at this concentration has no effect on Na+/K+ ATPase activity as well as cell ATP content. These findings suggest that stevioside, at a pharmacological concentration of 0.70 mM, inhibits transepithelial transport of PAH by interfering with the basolateral entry step, the rate-limiting step for transepithelial transport. The lack of effect of stevioside on transepithelial transport of PAH on the luminal side and its reversible inhibitory effect on the basolateral side indicate that stevioside does not permanently change PAH transport and should not harm renal tubular function at normal human intake levels. PMID- 11007538 TI - Inhibition of Ca2+-activated K+ channels by tyrosine phosphatase inhibitors in rat mesenteric artery. AB - To investigate the possible regulation of large-conductance Ca2+-activated K+ channels (BKCa) by tyrosine phosphatases (Tyr-PPs), single-channel currents of myocytes from rat mesenteric artery were recorded in open cell-attached patches. Two structurally different Tyr-PP inhibitors, sodium orthovanadate (Na3VO4) and dephostatin, were used. The channels (236 pS) evoked at +40 mV and pCa 6, were significantly inhibited by 1 mM Na3VO4 (-81+/-3%, n = 10; P < 0.005). Similarly, 100 microM dephostatin strongly inhibited the BKCa channels (-80+/-7%, n = 7 ; P < 0.05). Therefore, BKCa channels in vascular smooth muscle cells may be regulated by tyrosine phosphatase-dependent signal transduction pathways, whose inhibition could attenuate the channel activity. PMID- 11007539 TI - Block of inwardly rectifying K+ currents by extracellular Mg2+ and Ba2+ in bovine pulmonary artery endothelial cells. AB - Using whole-cell patch clamp technique, we investigated the blocking effects of extracellular Ba2+ and Mg2+ on the inwardly rectifying K+ (KIR) currents of bovine pulmonary artery endothelial cells (BPAEC). The BPAEC KIR channel has recently been identified as Kir2.1 of the Kir2.0 subfamily. Block of KIR currents by Mg2+ (3-30 mM) was instantaneous, and increased with hyperpolarization slightly (Kd at -160 and 0 mV was 9.5 and 23.2 mM, respectively). The apparent fractional electrical distance (delta) of the Mg2+ binding site is calculated to be 0.07 from the outer mouth of the channel pore. Ba2+ (0.3-10 microM) time dependently blocked the KIR currents with a much higher potency and stronger voltage-dependence (Kd at -160 and 0 mV was 1.0 and 41.6 microM, respectively). The Ba2+ binding site had a delta value of 0.34. Our data suggest that Mg2+ binds to a very superficial site of the KIR channel, while Ba2+ binds to a much deeper site, sensing much more of the membrane electric field. Thus, the BPAEC Kir2.1 appears to be pharmacologically different from the Kir2.1 reported before in bovine aortic endothelial cells (BAEC), which has 2 sites for Mg2+ block (a deep site in addition to a shallow one), and a superficial and low-sensitivity site for Ba2+ block. PMID- 11007540 TI - Variation in the vitreous phenotype of Stickler syndrome can be caused by different amino acid substitutions in the X position of the type II collagen Gly X-Y triple helix. AB - Stickler syndrome is a dominantly inherited disorder characterized by arthropathy, midline clefting, hearing loss, midfacial hypoplasia, myopia, and retinal detachment. These features are highly variable both between and within families. Mutations causing the disorder have been found in the COL2A1 and COL11A1 genes. Premature termination codons in COL2A1 that result in haploinsufficiency of type II collagen are a common finding. These produce a characteristic congenital "membranous" anomaly of the vitreous of all affected individuals. Experience has shown that vitreous slit-lamp biomicroscopy can distinguish between patients with COL2A1 mutations and those with dominant negative mutations in COL11A1, who produce a different "beaded" vitreous phenotype. Here we characterize novel dominant negative mutations in COL2A1 that result in Stickler syndrome. Both alter amino acids in the X position of the Gly X-Y triple-helical region. A recurrent R365C mutation occurred in two unrelated sporadic cases and resulted in the membranous vitreous anomaly associated with haploinsufficiency. In a large family with linkage to COL2A1, with a LOD score of 2.8, a unique L467F mutation produced a novel "afibrillar" vitreous gel devoid of all normal lamella structure. These data extend the mutation spectrum of the COL2A1 gene and help explain the basis for the different vitreous phenotypes seen in Stickler syndrome. PMID- 11007542 TI - Dendritic mechanisms in brain function and developmental disabilities. PMID- 11007541 TI - Small evolutionarily conserved RNA, resembling C/D box small nucleolar RNA, is transcribed from PWCR1, a novel imprinted gene in the Prader-Willi deletion region, which Is highly expressed in brain. AB - Prader-Willi syndrome is a complex neurodevelopmental disorder caused by the inactivation or deletion of imprinted, paternally expressed genes in chromosome band 15q11.2. We report the identification and characterization of PWCR1, a novel imprinted gene within that region, and its mouse orthologue, Pwcr1, which was mapped to the conserved syntenic region on mouse chromosome 7. Expressed only from the paternal allele, both genes require the imprinting-center regulatory element for expression and are transcribed from the same strand. They are intronless and do not appear to encode a protein product. High human/mouse sequence similarity (87% identity) is limited to a 99-bp region called "HMCR" (for "human-mouse conserved region"). The HMCR sequence has features of a C/D box small nucleolar RNA (snoRNA) and is represented in an abundant small transcript in both species. Located in nucleoli, snoRNAs serve as methylation guidance RNAs in the modification of ribosomal RNA and other small nuclear RNAs. In addition to the nonpolyadenylated small RNAs, larger polyadenylated PWCR1 transcripts are found in most human tissues, whereas expression of any Pwcr1 RNAs is limited to mouse brain. Genomic sequence analysis reveals the presence of multiple copies of PWCR1 and Pwcr1 that are organized within local tandem-repeat clusters. On a multispecies Southern blot, hybridization to an HMCR probe encoding the putative snoRNA is limited to mammals. PMID- 11007543 TI - Regulation of dendritic spine morphology by the rho family of small GTPases: antagonistic roles of Rac and Rho. AB - Dendritic spines mediate most excitatory transmission in the mammalian CNS and have been traditionally considered stable structures. Following the suggestion that spines may 'twitch', it has been recently shown that spines are capable of rapid morphological rearrangements. Because of the role of the small GTPases from the Rho family in controlling neuronal morphogenesis, we investigated the effects of several members of this biochemical signaling pathway in the maintenance of the morphology of extant dendritic spines by combining biolistic transfection of pyramidal neurons in cultured cortical and hippocampal slices with two-photon microscopy. We find a variety of effects on the density and morphology of dendritic spines by expressing either constitutively active or dominant negative forms of several small GTPases of the Rho family, by blocking the entire pathway with Clostridium difficile toxin B or by blocking Rho with C3 transferase. We propose a model where Rac promotes spine formation, while Rho prevents it. We conclude that the small GTPases provide antagonistic control mechanisms of spine maintenance in pyramidal neurons. PMID- 11007544 TI - Thyroid hormone action in neural development. AB - Effects of thyroid hormone on development of the brain have been documented for over a century. Although in many respects the hypothyroid brain appears morphologically normal, functional impairments include mental retardation, ataxia and spasticity. Keyed by the discovery of nuclear receptors for thyroid hormone that function as transcription factors, recent work has examined the mechanism of thyroid hormone action in brain development. The prediction that gene expression regulated by thyroid hormone is important for mediating brain development has spurred the search for thyroid hormone-responsive genes. Here we review some of the identified genes whose expression patterns correlate with the functional deficits observed in the hypothyroid brain. Recently identified thyroid hormone responsive genes include synaptotagmin-related gene 1 (Srg1), a putative mediator of synaptic structure and/or activity, and hairless, a transcriptional cofactor that may influence the expression of other thyroid hormone-responsive genes. PMID- 11007545 TI - Role of afferent innervation and neuronal activity in dendritic development and spine maturation of fascia dentata granule cells. AB - By using slice cultures of hippocampus as a model, we have studied the development of dendritic spines in fascia dentata granule cells. We raised the question as to what extent spine development is dependent on a major afferent input to these neurons, the fibers from the entorhinal cortex and neuronal activity mediated by these axons. Our results can be summarized as follows: (i) the entorhino-hippocampal projection develops in an organotypic manner in co cultures of entorhinal cortex and hippocampus. Like in vivo, entorhinal fibers, labeled by anterograde tracing with biocytin, terminate in the outer molecular layer of the fascia dentata. (ii) The layer-specific termination of entorhinal fibers is not altered by the blockade of neuronal activity with tetrodotoxin. Likewise, the differentiation of the dendritic arbor of postsynaptic granule cells does not require neuronal activity. Blockade of neuronal activity did not affect the mean spine number of granule cell dendrites in entorhino-hippocampal co-cultures, but led to a relative increase in thin, long filiform spines that are characteristic of immature neurons. (iii) The maturation of the granule cell dendritic arbor is, however, controlled by the afferent fibers from the entorhinal cortex in an activity-independent manner. In single slice cultures of hippocampus lacking entorhinal input, Golgi-impregnated granule cells have much shorter, less branched dendrites when compared with granule cells in entorhino hippocampal co-cultures. This reduction in dendritic length in granule cells lacking entorhinal input results in a lower mean total number of spines per neuron, but the mean number of spines per microm is not reduced in the absence of entorhinal innervation. These results indicate that innervation by fibers from the entorhinal cortex, but not neuronal activity mediated via these axons, is essential for the normal development of the granule cell dendritic arbor. Neuronal activity is required, however, for the maturation of dendritic spines. PMID- 11007546 TI - Structural synaptic modifications associated with hippocampal LTP and behavioral learning. AB - An important problem in the neurobiology of memory is whether cellular mechanisms of learning and memory include the formation of new synapses or the remodeling of existing ones. To elucidate this problem, numerous studies have examined alterations in the number and structure of synapses following behavioral learning and hippocampal long-term potentiation (LTP), which is viewed as a synaptic model of memory. The data reported in the literature and obtained in this laboratory are analyzed here to evaluate what kind of structural modification is likely to account for synaptic plasticity associated with learning and memory. It has been demonstrated that LTP induction elicits the formation of additional synapses between activated axon terminals and newly emerging dendritic spines. Similarly, some forms of learning have been shown to increase the number of synapses. Although many ultrastructural studies examining the effect of LTP or learning failed to find a change in total synapse number, this population measure might not detect an increase in a small proportion of synapses established by activated terminals. LTP and learning have also been shown to induce a remodeling of synapses. This process is proposed to involve the transformation of certain synaptic subtypes into more efficacious ones, including the conversion of 'silent' synapses into functional synapses. It appears, therefore, that cellular mechanisms of learning and memory are likely to include both synaptogenesis and synapse remodeling. PMID- 11007547 TI - Cellular and molecular mechanisms of dendrite growth. AB - Proper growth and branching of dendrites are crucial for nervous system function; patterns of dendritic arborization determine the nature and amount of innervation that a neuron receives and specific dendritic membrane properties define its computational capabilities. Until recently, there was relatively little known about the cellular and molecular mechanisms of dendritic growth, perhaps because dendrites were historically considered to be intrinsically determined, passive elements in the formation of connections in the nervous system. In the last few years, however, overwhelming evidence has accumulated indicating that dendritic growth is remarkably dynamic and responsive to environmental signals, including guidance molecules and levels and patterns of activity. This manuscript reviews our current understanding of the cellular and molecular mechanisms of dendritic growth, the influence of activity in sculpting specific patterns of dendritic arbors, and a potential integral role for dendrites in activity-dependent development of circuits in the nervous system. PMID- 11007548 TI - Serotonin induces EPSCs preferentially in layer V pyramidal neurons of the frontal cortex in the rat. AB - The effect of serotonin (5-HT) on the release of glutamate was examined in pyramidal cells in layers II-VI of the frontal cortex. The intracellular recording electrode contained 1% biocytin so the neurons could later be visualized with an avidin-biotin peroxidase method. Pyramidal cells in layer V of the frontal cortex showed the greatest 5-HT-induced increase in both the frequency and amplitude of 'spontaneous' (non-electrically evoked) excitatory post-synaptic currents (EPSCs). A small proportion of neurons in layer II/III showed an increase in EPSC frequency, whereas cells in layer VI showed no significant change in either EPSC frequency or amplitude. The physiological response to 5-HT mirrors the high density of 5-HT(2A) receptors in layer V, as well as the pattern of thalamic projections in frontal cortex. The specific induction of EPSCs in layer V neurons suggests that 5-HT preferentially modulates the output neurons of the frontal cortex. PMID- 11007549 TI - Dendritic anomalies in disorders associated with mental retardation. AB - Dendritic abnormalities are the most consistent anatomical correlates of mental retardation (MR). Earliest descriptions included dendritic spine dysgenesis, which was first associated with unclassified MR, but can also be found in genetic syndromes associated with MR. Genetic disorders with well-defined dendritic anomalies involving branches and/or spines include Down, Rett and fragile-X syndromes. Cytoarchitectonic analyses also suggest dendritic pathology in Williams and Rubinstein-Taybi syndromes. Dendritic abnormalities appear to have syndrome-specific pathogenesis and evolution, which correlate to some extent with their cognitive profile. The significance of dendritic pathology in synaptic circuitry and the role of animal models in the study of MR-associated dendritic abnormalities are also discussed. Finally, a model of genotype to neurologic phenotype pathway in MR, centered in dendritic abnormalities, is postulated. PMID- 11007550 TI - Dendritic cytoskeletal protein expression in mental retardation: an immunohistochemical study of the neocortex in Rett syndrome. AB - Many syndromes associated with mental retardation (MR) are characterized by cortical dendritic anomalies. Despite their morphological similarity, these changes appear to involve different stages of dendritic development. The neuronal cytoskeleton, which includes microfilaments, neurofilaments and microtubules, is essential for these developmental processes. Levels and phosphorylation of microtubule-associated proteins (MAPs), which stabilize microtubules, seem to determine different stages of dendritic formation with certain MAPs (e.g. MAP-2) appearing to mediate the effects of external modulators upon these processes. Early studies on neuronal cytoskeleton in MR, which have shown a selective reduction in MAP-2 expression, have focused on Rett syndrome (RS). Here, by a semiquantitative immunohistochemical analysis of the pericentral cortex, we examine the contribution of specific neuronal populations to these changes in cytoskeletal proteins. Decreased MAP-2 staining in RS was more marked in layers V VI, while increased nonphosphorylated neurofilament immunoreactivity was found in layers II-III in RS. Age-related increases in dendritic MAP-2 immunoreactivity in layers V-VI were also absent in RS. The specificity of these cytoskeletal protein changes, their significance for RS pathogenesis and plasticity, as well as their implications for other MR-associated disorders, are also discussed. PMID- 11007551 TI - Connectivity of ectopic neurons in the molecular layer of the somatosensory cortex in autoimmune mice. AB - Approximately 50% of New Zealand Black mice (NZB/BINJ) and 80% of NXSM-D/EiJ mice prenatally develop neocortical layer I ectopias, mostly in somatosensory cortices. These cortical anomalies are similar to those seen in the brains of individuals with dyslexia. Neurofilament staining revealed a radial column of tightly packed fiber bundles in the layers underlying ectopias. This suggested that the connectivity of the ectopic neurons was aberrant. The present study used the tracers 1,1'-dioctadecyl- 3,3,3',3'-tetramethylindo- carbocyanine perchlorate (DiI) and biotinylated dextran amine (BDA) to more thoroughly explore the cortical and thalamic connectivity of the ectopias. DiI placement into ectopias again revealed a distinct bundle of fibers extending from the ectopic neurons to the deep cortical layers. This bundle split in the white matter with some fibers traveling to the corpus callosum and others to the internal capsule. Thalamic connections were concentrated in the ventrobasal com- plex (VB) and posterior thalamic nucleus group (Po). Injections of BDA into VB revealed reciprocal connections between VB and the ectopic cortical neurons. Ipsilateral corticocortical projections were seen between ectopias in primary somatosensory and motor and secondary somatosensory cortices, but no contralateral connections of the ectopic neurons were seen. These findings confirm the notion that layer I ectopias are anomalously connected by comparison to neurons in homologous cortex, which may underlie widespread dysfunction of brains containing ectopias. PMID- 11007552 TI - Convergence and plasticity of monoaminergic systems in the medial prefrontal cortex during the postnatal period: implications for the development of psychopathology. AB - A variety of observations have suggested that the dopamine and serotonin systems may play a role in the pathophysiology and treatment of major mental disorders of childhood, adolescence and early adulthood. A recent triple immunofluorescence study has demonstrated a convergence of serotonin and dopamine fibers onto both pyramidal cells and GABAergic interneurons in the rat medial prefrontal cortex (mPFCx). These findings are consistent with the results of an electrophysiological study conducted in another laboratory that suggested such a relationship exists in the pyriform cortex of the rodent brain. During postnatal development, the dopamine system shows a progressive ingrowth of fibers into this region that continues until the early adult period. In contrast, GABAergic neurons appear to complete their postnatal maturation by the fourth postnatal week (the early post-weanling period). As dopamine fibers infiltrate the rat mPFCx, they progressively increase their interaction with neural elements within the neuropil and with the cell bodies of both pyramidal cells and GABAergic interneurons. This process appears to be influenced by the serotonin system, since lesioning of the nucleus raphe dorsalis during the neonatal period results in a significant increase of dopamine fibers. This finding suggests that lesions of the serotonin system induce plasticity of the cortical dopamine system; however, it is not known whether this inferred suppressive effect of serotonin fibers occurs at brainstem levels or within the mPFCx itself. Taken together, these various studies suggest that the convergence of dopamine and serotonin fiber systems on intrinsic cortical neurons shows considerable plasticity during postnatal life that could theoretically contribute to the development of 'miswired' circuits in individuals with neuropsychiatric disorders. PMID- 11007553 TI - Gangliosides as modulators of dendritogenesis in normal and storage disease affected pyramidal neurons. AB - Pyramidal cells initiate the formation of dendritic arbors in a prolific burst of neurite outgrowth during early cortical development. Although morphologically mature pyramidal neurons do not normally sprout additional primary dendrites, the discovery of ectopic dendritogenesis in neuronal storage diseases has revealed that these cells do retain this ability under appropriate stimulation. The capacity for renewal of dendritogenesis has been found to exhibit a species gradient with human > cat, dog, sheep > mouse. A consistent metabolic feature of ectopic dendrite-bearing pyramidal neurons is a heightened intracellular expression of GM2 ganglioside. Elevated expression of this same glycosphingolipid has also been found to correlate with normal dendritogenesis. Immature neurons in developing cat and ferret cortex exhibit high levels of GM2 ganglioside immunoreactivity coincident with normal dendritic sprouting and a similar relationship has now been shown for human cortical development. Ultrastructural studies of all three species revealed GM2 localized to vesicles in a manner consistent with Golgi synthesis and exocytic trafficking to the somatic-dendritic plasmalemma. We propose that GM2 ganglioside functions in glycosphingolipid enriched microdomains (lipid rafts) in the plasmalemma to promote dendritic initiation through modulation of specific membrane proteins and/or their associated second messenger cascades. PMID- 11007554 TI - Dendritic spine structural anomalies in fragile-X mental retardation syndrome. AB - Fragile-X syndrome is the most common single-gene inherited form of mental retardation. Morphological studies suggest a possible failure of the synapse maturation process. Cerebral cortical spine morphology in fragile-X syndrome and in a knockout mouse model of it appears immature, with long, thin spines much more common than the stubby and mushroom-shaped spines more characteristic of normal development. In human fragile-X syndrome there is also a higher density of spines along dendrites, suggesting a possible failure of synapse elimination. While variously misshapen spines are characteristic of a number of mental retardation syndromes, the overabundance of spines seen in fragile-X syndrome is unusual. Taken with evidence of neurotransmitter activation of the synthesis of the fragile-X protein (FMRP) at synapses in vitro and evidence for behaviorally induced FMRP expression in vivo, and with evidence compatible with a role for FMRP in regulating the synthesis of other proteins, it is possible that FMRP serves as an 'immediate early protein' at the synapse that orchestrates aspects of synaptic development and plasticity. PMID- 11007555 TI - FMRP involvement in formation of synapses among cultured hippocampal neurons. AB - Fragile-X, the main cause of inherited human mental retardation is associated with the absence of a recently identified fragile-X mental retardation protein (FMRP). Mice in which this protein is lacking due to a knockout (KO) mutation are reported to express altered dendritic spines on their cortical neurons compared with wild type (WT) controls. We have used tissue-cultured neurons to examine differences in morphology and synaptic connectivity between WT and FMRP-deficient mice. Hippocampal neurons taken from KO mice and grown in culture for 3 weeks have shorter dendrites and fewer dendritic spines than their WT counterparts. Also, KO cells tend to express fewer functional synaptic connections, which develop more slowly and produce smaller excitatory synaptic currents than WT controls. These observations may have important implications for the understanding of mental retardation associated with the absence of FMRP. PMID- 11007556 TI - Highlighted topics series: cellular responses to mechanical stress. PMID- 11007557 TI - Beta-adrenergic agonist therapy accelerates the resolution of hydrostatic pulmonary edema in sheep and rats. AB - To determine whether beta-adrenergic agonist therapy increases alveolar liquid clearance during the resolution phase of hydrostatic pulmonary edema, we studied alveolar and lung liquid clearance in two animal models of hydrostatic pulmonary edema. Hydrostatic pulmonary edema was induced in sheep by acutely elevating left atrial pressure to 25 cmH(2)O and instilling 6 ml/kg body wt isotonic 5% albumin (prepared from bovine albumin) in normal saline into the distal air spaces of each lung. After 1 h, sheep were treated with a nebulized beta-agonist (salmeterol) or nebulized saline (controls), and left atrial pressure was then returned to normal. beta-Agonist therapy resulted in a 60% increase in alveolar liquid clearance over 3 h (P < 0.001). Because the rate of alveolar fluid clearance in rats is closer to human rates, we studied beta-agonist therapy in rats, with hydrostatic pulmonary edema induced by volume overload (40% body wt infusion of Ringer lactate). beta-Agonist therapy resulted in a significant decrease in excess lung water (P < 0.01) and significant improvement in arterial blood gases by 2 h (P < 0.03). These preclinical experimental studies support the need for controlled clinical trials to determine whether beta-adrenergic agonist therapy would be of value in accelerating the resolution of hydrostatic pulmonary edema in patients. PMID- 11007558 TI - Increased spinal monoamine concentrations after chronic thoracic dorsal rhizotomy in goats. AB - In goats, bilateral thoracic dorsal rhizotomy (TDR) causes severe ventilatory failure during exercise, followed by progressive functional recovery. We investigated spinal neurochemical changes associated with TDR and/or functional recovery by measuring spinal concentrations of the monoamines serotonin (5-HT), norepinephrine, and dopamine via HPLC. Changes in 5-HT and calcitonin gene related peptide were visualized with immunohistochemistry. Goat spinal cords were compared 4-15 mo after TDR from T(2) to T(12) (n = 7) with sham-operated (n = 4) or unoperated controls (n = 4). TDR increased the concentration of cervical 5-HT (C(5)-C(6); 122% change), caudal thoracic norepinephrine (T(7)-T(11); 53% change), and rostral thoracic dopamine (T(3)-T(6); 234% change). TDR increased 5 HT-immunoreactive terminal density (dorsal and ventral horns) and nearly eliminated calcitonin gene-related peptide immunoreactivity in the superficial laminae of the dorsal horn in rostral thoracic segments; both effects became less pronounced in caudal thoracic segments. Thus TDR elevates monoamine concentrations in discrete spinal regions, including possible compensatory changes in descending serotonergic inputs to spinal segments not directly affected by TDR (i.e., cervical) but associated with functionally related motor nuclei (i.e., phrenic nucleus). PMID- 11007559 TI - Upper airway muscle responsiveness to rising PCO(2) during NREM sleep. AB - Although pharyngeal muscles respond robustly to increasing PCO(2) during wakefulness, the effect of hypercapnia on upper airway muscle activation during sleep has not been carefully assessed. This may be important, because it has been hypothesized that CO(2)-driven muscle activation may importantly stabilize the upper airway during stages 3 and 4 sleep. To test this hypothesis, we measured ventilation, airway resistance, genioglossus (GG) and tensor palatini (TP) electromyogram (EMG), plus end-tidal PCO(2) (PET(CO(2))) in 18 subjects during wakefulness, stage 2, and slow-wave sleep (SWS). Responses of ventilation and muscle EMG to administered CO(2) (PET(CO(2)) = 6 Torr above the eupneic level) were also assessed during SWS (n = 9) or stage 2 sleep (n = 7). PET(CO(2)) increased spontaneously by 0.8 +/- 0.1 Torr from stage 2 to SWS (from 43.3 +/- 0.6 to 44.1 +/- 0.5 Torr, P < 0.05), with no significant change in GG or TP EMG. Despite a significant increase in minute ventilation with induced hypercapnia (from 8.3 +/- 0.1 to 11.9 +/- 0.3 l/min in stage 2 and 8.6 +/- 0.4 to 12.7 +/- 0.4 l/min in SWS, P < 0.05 for both), there was no significant change in the GG or TP EMG. These data indicate that supraphysiological levels of PET(CO(2)) (50.4 +/- 1.6 Torr in stage 2, and 50.4 +/- 0.9 Torr in SWS) are not a major independent stimulus to pharyngeal dilator muscle activation during either SWS or stage 2 sleep. Thus hypercapnia-induced pharyngeal dilator muscle activation alone is unlikely to explain the paucity of sleep-disordered breathing events during SWS. PMID- 11007560 TI - Altered airway surfactant phospholipid composition and reduced lung function in asthma. AB - Pulmonary surfactant in bronchoalveolar lavage fluid (BALF) and induced sputum from adults with stable asthma (n = 36) and healthy controls (n = 12) was analyzed for phospholipid and protein compositions and function. Asthmatic subjects were graded as mild, moderate, or severe. Phospholipid compositions of BALF and sputum from control subjects were similar and characteristic of surfactant. For asthmatic subjects, the proportion of dipalmitoyl phosphatidylcholine (16:0/16:0PC), the major phospholipid in surfactant, decreased in sputum (P < 0.05) but not in BALF. In BALF, mole percent 16:0/16:0PC correlated with surfactant function measured in a capillary surfactometer, and sputum mole percent 16:0/16:0PC correlated with lung function (forced expiratory volume in 1 s). Neither surfactant protein A nor total protein concentration in either BALF or sputum was altered in asthma. These results suggest altered phospholipid composition and function of airway (sputum) but not alveolar (BALF) surfactant in stable asthma. Such underlying surfactant dysfunction may predispose asthmatic subjects to further surfactant inhibition by proteins or aeroallergens in acute asthma episodes and contribute to airway closure in asthma. Consequently, administration of an appropriate therapeutic surfactant could provide clinical benefit in asthma. PMID- 11007562 TI - Postexercise rehydration: effect of Na(+) and volume on restoration of fluid spaces and cardiovascular function. AB - Our purpose was to study the interaction between Na(+) content and fluid volume on rehydration (RH) and restoration of fluid spaces and cardiovascular (CV) function. Ten men completed four trials in which they exercised in a 35 degrees C environment until dehydrated by 2. 9% body mass, were rehydrated for 180 min, and exercised for an additional 20 min. Four RH regimens were tested: low volume (100% fluid replacement)-low (25 mM) Na(+) (LL), low volume-high (50 mM) Na(+) (LH), high volume (150% fluid replacement)-low Na(+) (HL), and high volume-high Na(+) (HH). Blood and urine samples were collected and body mass was measured before and after exercise and every hour during RH. Before and after the dehydration exercise and during the 20 min of exercise after RH, cardiac output was measured. Fluid compartment (intracellular and extracellular) restoration and percent change in plasma volume were calculated using the Cl(-) and hematocrit/Hb methods, respectively. RH was greater (P < 0.05) in HL and HH (102.0 +/- 15.2 and 103.7 +/- 14.7%, respectively) than in LL and LH (70.7 +/- 10.5 and 75.9 +/- 6.3%, respectively). Intracellular RH was greater in HL (1.12 +/- 0.4 liters) than in all other conditions (0.83 +/- 0.3, 0.69 +/- 0.2, and 0.73 +/- 0.3 liter for LL, LH, and HH, respectively), whereas extracellular RH (including plasma volume) was greater in HL and HH (1.35 +/- 0.8 and 1.63 +/- 0.4 liters, respectively) than in LL and LH (0.83 +/- 0.3 and 1.05 +/- 0.4 liters, respectively). CV function (based on stroke volume, heart rate, and cardiac output) was restored equally in all conditions. These data indicate that greater RH can be achieved through larger volumes of fluid and is not affected by Na(+) content within the range tested. Higher Na(+) content favors extracellular fluid filling, whereas intracellular fluid benefits from higher volumes of fluid with lower Na(+). Alterations in Na(+) and/or volume within the range tested do not affect the degree of restoration of CV function. PMID- 11007561 TI - Role of convective O(2) delivery in determining VO(2) on-kinetics in canine muscle contracting at peak VO(2). AB - A previous study (Grassi B, Gladden LB, Samaja M, Stary CM, and Hogan MC, J Appl Physiol 85: 1394-1403, 1998) showed that convective O(2) delivery to muscle did not limit O(2) uptake (VO(2)) on-kinetics during transitions from rest to contractions at approximately 60% of peak VO(2). The present study aimed to determine whether this finding is also true for transitions involving contractions of higher metabolic intensities. VO(2) on-kinetics were determined in isolated canine gastrocnemius muscles in situ (n = 5) during transitions from rest to 4 min of electrically stimulated isometric tetanic contractions corresponding to the muscle peak VO(2). Two conditions were compared: 1) spontaneous adjustment of muscle blood flow (Q) (Control) and 2) pump-perfused Q, adjusted approximately 15-30 s before contractions at a constant level corresponding to the steady-state value during contractions in Control (Fast O(2) Delivery). In Fast O(2) Delivery, adenosine was infused intra-arterially. Q was measured continuously in the popliteal vein; arterial and popliteal venous O(2) contents were measured at rest and at 5- to 7-s intervals during the transition. Muscle VO(2) was determined as Q times the arteriovenous blood O(2) content difference. The time to reach 63% of the VO(2) difference between resting baseline and steady-state values during contractions was 24.9 +/- 1.6 (SE) s in Control and 18.5 +/- 1.8 s in Fast O(2) Delivery (P < 0.05). Faster VO(2) on kinetics in Fast O(2) Delivery was associated with an approximately 30% reduction in the calculated O(2) deficit and with less muscle fatigue. During transitions involving contractions at peak VO(2), convective O(2) delivery to muscle, together with an inertia of oxidative metabolism, contributes in determining the VO(2) on-kinetics. PMID- 11007563 TI - Differences in airway structure in immature and mature rabbits. AB - Our laboratory has previously demonstrated that maximal bronchoconstriction produces a greater degree of airway narrowing in immature than in mature rabbit lungs (33). To determine whether these maturational differences could be related to airway structure, we compared the fraction of the airway wall occupied by airway smooth muscle (ASM) and cartilage, the proportion of wall area internal to ASM, and the number of alveolar attachments to the airways, from mature and immature (6-mo- and 4-wk-old, respectively) rabbit lungs that were formalin fixed at total lung capacity. The results demonstrate that the airway walls of immature rabbits had a greater percentage of smooth muscle, a lower percentage of cartilage, and fewer alveolar attachments compared with mature rabbit airways; however, we did not find maturational differences in the airway wall thickness relative to airway size. We conclude that structural differences in the airway wall may contribute to the greater airway narrowing observed in immature rabbits during bronchoconstriction. PMID- 11007564 TI - Venoarterial CO(2) difference during regional ischemic or hypoxic hypoxia. AB - To test the role of blood flow in tissue hypoxia-related increased veno-arterial PCO(2) difference (DeltaPCO(2)), we decreased O(2) delivery (&Ddot;O(2)) by either decreasing flow [ischemic hypoxia (IH)] or arterial PO(2) [hypoxic hypoxia (HH)] in an in situ, vascularly isolated, innervated dog hindlimb perfused with a pump-membrane oxygenator system. Twelve anesthetized and ventilated dogs were studied, with systemic hemodynamics maintained within normal range. In the IH group (n = 6), hindlimb DO(2) was progressively lowered every 15 min by decreasing pump-controlled flow from 60 to 10 ml. kg(-1). min(-1), with arterial PO(2) constant at 100 Torr. In the HH group (n = 6), hindlimb DO(2) was progressively lowered every 15 min by decreasing PO(2) from 100 to 15 Torr, when flow was constant at 60 ml. kg(-1). min(-1). Limb DO(2), O(2) uptake (VO(2)), and DeltaPCO(2) were obtained every 15 min. Below the critical DO(2), VO(2) decreased, indicating dysoxia, and O(2) extraction ratio (VO(2)/DO(2)) rose continuously and similarly in both groups, reaching a maximal value of approximately 90%. DeltaPCO(2) significantly increased in IH but never differed from baseline in HH. We conclude that absence of increased DeltaPCO(2) does not preclude the presence of tissue dysoxia and that decreased flow is a major determinant in increased DeltaPCO(2). PMID- 11007565 TI - A predictive model of fatigue in human skeletal muscles. AB - Fatigue is a major limitation to the clinical application of functional electrical stimulation. The activation pattern used during electrical stimulation affects force and fatigue. Identifying the activation pattern that produces the greatest force and least fatigue for each patient is, therefore, of great importance. Mathematical models that predict muscle forces and fatigue produced by a wide range of stimulation patterns would facilitate the search for optimal patterns. Previously, we developed a mathematical isometric force model that successfully identified the stimulation patterns that produced the greatest forces from healthy subjects under nonfatigue and fatigue conditions. The present study introduces a four-parameter fatigue model, coupled with the force model that predicts the fatigue induced by different stimulation patterns on different days during isometric contractions. This fatigue model accounted for 90% of the variability in forces produced by different fatigue tests. The predicted forces at the end of fatigue testing differed from those observed by only 9%. This model demonstrates the potential for predicting muscle fatigue in response to a wide range of stimulation patterns. PMID- 11007566 TI - Neurocirculatory consequences of intermittent asphyxia in humans. AB - We examined the neurocirculatory and ventilatory responses to intermittent asphyxia (arterial O(2) saturation = 79-85%, end-tidal PCO(2) =3-5 Torr above eupnea) in seven healthy humans during wakefulness. The intermittent asphyxia intervention consisted of 20-s asphyxic exposures alternating with 40-s periods of room-air breathing for a total of 20 min. Minute ventilation increased during the intermittent asphyxia period (14.2 +/- 2.0 l/min in the final 5 min of asphyxia vs. 7.5 +/- 0.4 l/min in baseline) but returned to the baseline level within 2 min after completion of the series of asphyxic exposures. Muscle sympathetic nerve activity increased progressively, reaching 175 +/- 12% of baseline in the final 5 min of the intervention. Unlike ventilation, sympathetic activity remained elevated for at least 20 min after removal of the chemical stimuli (150 +/- 10% of baseline in the last 5 min of the recovery period). Intermittent asphyxia caused a small, but statistically significant, increase in heart rate (64 +/- 4 beats/min in the final 5 min of asphyxia vs. 61 +/- 4 beats/min in baseline); however, this increase was not sustained after the return to room-air breathing. These data demonstrate that relatively short-term exposure to intermittent asphyxia causes sympathetic activation that persists after removal of the chemical stimuli. This carryover effect provides a potential mechanism whereby intermittent asphyxia during sleep could lead to chronic sympathetic activation in patients with sleep apnea syndrome. PMID- 11007567 TI - Effects of beta-hydroxy-beta-methylbutyrate on muscle damage after a prolonged run. AB - This study examined the effects of supplemental beta-hydroxy-beta-methylbutyrate (HMB) on muscle damage as a result of intense endurance exercise. Subjects (n = 13) were paired according to their 2-mile run times and past running experience. Each pair was randomly assigned a treatment of either HMB (3 g/day) or a placebo. After 6 wk of daily training and supplementation, all subjects participated in a prolonged run (20-km course). Creatine phosphokinase and lactate dehydrogenase (LDH) activities were measured before and after a prolonged run to assess muscle damage. The placebo-supplemented group exhibited a significantly greater (treatment main effect, P = 0.05) increase in creatine phosphokinase activity after a prolonged run than did the HMB-supplemented group. In addition, LDH activity was significantly lower (treatment main effect, P = 0.003) with HMB supplementation compared with the placebo-supplemented group. In conclusion, supplementation with 3.0 g of HMB results in a decreased creatine phosphokinase and LDH response after a prolonged run. These findings support the hypothesis that HMB supplementation helps prevent exercise-induced muscle damage. PMID- 11007568 TI - No evidence for long-term facilitation after episodic hypoxia in spontaneously breathing, anesthetized rats. AB - Repeated electrical or hypoxic stimulation of peripheral chemoreceptors has been shown to cause a persistent poststimulus increase in respiratory motoneuron activity, termed long-term facilitation (LTF). LTF after episodic hypoxia has been demonstrated most consistently in anesthetized, vagotomized, paralyzed, artificially ventilated rats. Evidence for LTF in spontaneously breathing animals and humans after episodic hypoxia is equivocal and may have been influenced by the awake state of the subjects in these studies. The present study was designed to test the hypothesis that LTF is evoked in respiratory-related tongue muscle and inspiratory pump muscle activities after episodic hypoxia in 10 spontaneously breathing, anesthetized, vagotomized rats. The animals were exposed to three (5 min) episodes of isocapnic hypoxia, separated by 5 min of hyperoxia (50% inspired oxygen). Genioglossus, hyoglossus, and inspiratory intercostal EMG activities, along with respiratory-related tongue movements and esophageal pressure, were recorded before, during, and for 60 min after the end of episodic isocapnic hypoxia. We found no evidence for LTF in tongue muscle (genioglossus, hyoglossus) or inspiratory pump muscle (inspiratory intercostal) activities after episodic hypoxia. Rather, the primary poststimulus effect of episodic hypoxia was diminished respiratory frequency, which contributed to a reduction in ventilatory drive. PMID- 11007569 TI - Dynamic viscoelastic behavior of lower extremity tendons during simulated running. AB - The aim of this project was to see whether the tendon would show creep during long-term dynamic loading (here referred to as dynamic creep). Pig tendons were loaded by a material-testing machine with a human Achilles tendon force profile (1.37 Hz, 3% strain, 1,600 cycles), which was obtained in an earlier in vivo experiment during running. All the pig tendons showed some dynamic creep during cyclic loading (between 0.23 +/- 0.15 and 0.42 +/- 0.21%, means +/- SD). The pig tendon data were used as an input of a model to predict dynamic creep in the human Achilles tendon during running of a marathon and to evaluate whether there might consequently be an influence on group Ia afferent-mediated length and velocity feedback from muscle spindles. The predicted dynamic creep in the Achilles tendon was considered to be too small to have a significant influence on the length and velocity feedback from soleus during running. In spite of the characteristic nonlinear viscoelastic behavior of tendons, our results demonstrate that these properties have a minor effect on the ability of tendons to act as predictable, stable, and elastic force transmitters during long-term cyclic loading. PMID- 11007570 TI - Differential effects of furosemide on porcine bronchial arterial and airway smooth muscle. AB - Furosemide attenuates airway obstruction in asthmatic subjects when administered as an aerosol pretreatment. This protective effect of furosemide could be related to relaxation of bronchial smooth muscle or to increased bronchial blood flow. To determine whether furosemide dilates bronchial smooth muscle, isometric contractile responses in distal bronchi from young pigs were studied. In bronchial smooth muscle rings that were precontracted with 10(-5) M acetylcholine, significant relaxation occurred with 10(-8) to 3 x 10(-6) M isoproterenol but not with 10(-8) to 10(-3) M furosemide. In contrast, bronchial arteries that were precontracted with either 10(-4) M norepinephrine or 10(-8) M vasopressin significantly relaxed in response to 10(-4) to 3 x 10(-3) M and 10( 3) to 3 x 10(-3) M furosemide, respectively. We conclude that furosemide, under the described experimental conditions, relaxes airway vascular smooth muscle but not bronchial smooth muscle. These results are consistent with previous suggestions that inhaled furosemide increases blood flow to airway tissues (Gilbert IA, Lenner KA, Nelson JA, Wolin AD, and Fouke JM. J Appl Physiol 76: 409 415, 1994). PMID- 11007571 TI - IGF-I restores satellite cell proliferative potential in immobilized old skeletal muscle. AB - One of the key factors responsible for the age-associated reduction in muscle mass may be that satellite cell proliferation potential (number of doublings contained within each cell) could become rate limiting to old muscle regrowth. No studies have tested whether repeated cycles of atrophy-regrowth in aged animals deplete the remaining capacity of satellite cells to replicate or what measures can be taken to prevent this from happening. We hypothesized that there would be a pronounced loss of satellite cell proliferative potential in gastrocnemius muscles of aged rats (25- to 30-mo-old FBN rats) subjected to three cycles of atrophy by hindlimb immobilization (plaster casts) with intervening recovery periods. Our results indicated that there was a significant loss in gastrocnemius muscle mass and in the proliferative potential of the resident satellite cells after just one bout of immobilization. Neither the muscle mass nor the satellite cell proliferation potential recovered from their atrophied values after either the first 3-wk or later 9-wk recovery period. Remarkably, application of insulin like growth factor I onto the atrophied gastrocnemius muscle for an additional 2 wk after this 9-wk recovery period rescued approximately 46% of the lost muscle mass and dramatically increased proliferation potential of the satellite cells from this muscle. PMID- 11007572 TI - Lower limb skeletal muscle mass: development of dual-energy X-ray absorptiometry prediction model. AB - Although magnetic resonance imaging (MRI) can accurately measure lower limb skeletal muscle (SM) mass, this method is complex and costly. A potential practical alternative is to estimate lower limb SM with dual-energy X-ray absorptiometry (DXA). The aim of the present study was to develop and validate DXA-SM prediction equations. Identical landmarks (i.e., inferior border of the ischial tuberosity) were selected for separating lower limb from trunk. Lower limb SM was measured by MRI, and lower limb fat-free soft tissue was measured by DXA. A total of 207 adults (104 men and 103 women) were evaluated [age 43 +/- 16 (SD) yr, body mass index (BMI) 24.6 +/- 3.7 kg/m(2)]. Strong correlations were observed between lower limb SM and lower limb fat-free soft tissue (R(2) = 0.89, P < 0.001); age and BMI were small but significant SM predictor variables. In the cross-validation sample, the differences between MRI-measured and DXA-predicted SM mass were small (-0.006 +/- 1.07 and -0.016 +/- 1.05 kg) for two different proposed prediction equations, one with fat-free soft tissue and the other with added age and BMI as predictor variables. DXA-measured lower limb fat-free soft tissue, along with other easily acquired measures, can be used to reliably predict lower limb skeletal muscle mass. PMID- 11007573 TI - Effects of prior heavy exercise on phase II pulmonary oxygen uptake kinetics during heavy exercise. AB - We tested the hypothesis that heavy-exercise phase II oxygen uptake (VO(2)) kinetics could be speeded by prior heavy exercise. Ten subjects performed four protocols involving 6-min exercise bouts on a cycle ergometer separated by 6 min of recovery: 1) moderate followed by moderate exercise; 2) moderate followed by heavy exercise; 3) heavy followed by moderate exercise; and 4) heavy followed by heavy exercise. The VO(2) responses were modeled using two (moderate exercise) or three (heavy exercise) independent exponential terms. Neither moderate- nor heavy intensity exercise had an effect on the VO(2) kinetic response to subsequent moderate exercise. Although heavy-intensity exercise significantly reduced the mean response time in the second heavy exercise bout (from 65.2 +/- 4.1 to 47.0 +/- 3.1 s; P < 0.05), it had no significant effect on either the amplitude or the time constant (from 23.9 +/- 1.9 to 25.3 +/- 2.9 s) of the VO(2) response in phase II. Instead, this "speeding" was due to a significant reduction in the amplitude of the VO(2) slow component. These results suggest phase II VO(2) kinetics are not speeded by prior heavy exercise. PMID- 11007574 TI - Inhaled porcine pancreatic elastase causes bronchoconstriction via a bradykinin mediated mechanism. AB - Neutrophil elastase has been linked to inflammatory lung diseases such as chronic obstructive pulmonary disease, adult respiratory distress syndrome, emphysema, and cystic fibrosis. In guinea pigs, aerosol challenge with human neutrophil elastase causes bronchoconstriction, but the mechanism by which this occurs is not completely understood. Our laboratory previously showed that human neutrophil elastase releases tissue kallikrein (TK) from cultured tracheal gland cells. TK has been identified as the major kininogenase of the airway and cleaves both high and low-molecular weight kininogen to yield lysyl-bradykinin. Because inhaled bradykinin causes bronchoconstriction and airway hyperresponsiveness in asthmatic patients and allergic sheep, we hypothesized that elastase-induced bronchoconstriction could be mediated by bradykinin. To test this hypothesis, we measured lung resistance (RL) in sheep before and after inhalation of porcine pancreatic elastase (PPE) alone and after pretreatment with a bradykinin B(2) antagonist (NPC-567), the specific human elastase inhibitor ICI 200,355, the histamine H(1)-antagonist diphenhydramine hydrochloride, the cysteinyl leukotriene 1 receptor antagonist montelukast, or the cyclooxygenase inhibitor indomethacin. Inhaled PPE (125-1,000 microg) caused a dose-dependent increase in RL. Aerosol challenge with a single 500 microg dose of PPE increased RL by 132 +/ 8% over baseline. This response was blocked by pretreatment with NPC-567 and ICI 200,355 (n = 6; P < 0.001), whereas treatment with diphenhydramine hydrochloride, montelukast, or indomethacin failed to block the PPE-induced bronchoconstriction. Consistent with pharmacological data, TK activity in bronchial lavage fluid increased 134 +/- 57% over baseline (n = 5; P < 0.02). We conclude that, in sheep, PPE-induced bronchoconstriction is in part mediated by the generation of bradykinin. Our findings suggest that elastase-kinin interactions may contribute to changes in bronchial tone during inflammatory diseases of the airways. PMID- 11007575 TI - Comparison of thermoregulatory responses between men and women immersed in cold water. AB - Eleven women (age = 24.4 +/- 6.3 yr, mass = 65.0 +/- 7.8 kg, height = 167 +/- 8 cm, body fatness = 22.4 +/- 5.9%, mean +/- SD) were immersed to neck level in 18 degrees C water for up to 90 min for comparison of their thermal responses with those of men (n = 14) in a previous similarly conducted protocol. Metabolic rate increased about three times resting levels in men and women, whereas the rate of rectal temperature cooling (DeltaT(re)/Deltat) in women (0.47 degrees C/h) was about one-half that in men. With use of all data, DeltaT(re)/Deltat correlates with the ratio of body surface area to size and the metabolic rate of shivering correlates inversely to the square root of body fatness. No significant gender differences in total metabolic heat production normalized for body mass or surface area were found among subjects who completed 90 min of immersion (9 women and 7 men). Nor was there a gender difference in the overall percent contribution ( approximately 60%) of fat oxidation to total heat production. Blood concentrations of free fatty acids, glycerol, beta-hydroxybutyrate, and lactate increased significantly during the 90-min immersion, whereas muscle glycogen sampled from the right quadriceps femoris vastus lateralis decreased (free fatty acids, glycerol, and beta-hydroxybutyrate were higher in women). When the subjects were subgrouped according to similar body fatness and 60 min of immersion (6 women and 5 men), no significant gender differences emerged in DeltaT(re)/Deltat, energy metabolism, and percent fat oxidation. These findings suggest that no gender adjustments are necessary for prediction models of cold response if body fatness and the ratio of body surface area to size are taken into account and that a potential gender advantage with regard to carbohydrate sparing during cold water immersion is not supported. PMID- 11007576 TI - Insulin and exercise differentially regulate PI3-kinase and glycogen synthase in human skeletal muscle. AB - The purpose of this study was to determine the separate and combined effects of exercise and insulin on the activation of phosphatidylinositol 3-kinase (PI3 kinase) and glycogen synthase in human skeletal muscle in vivo. Seven healthy men performed three trials in random order. The trials included 1) ingestion of 2 g/kg body wt carbohydrate in a 10% solution (CHO); 2) 75 min of semirecumbent cycling exercise at 75% of peak O(2) consumption; followed by 5 x 1-min maximal sprints (Ex); and 3) Ex, immediately followed by ingestion of the carbohydrate solution (ExCHO). Plasma glucose and insulin were increased (P < 0.05) at 15 and 30 (Post-15 and Post-30) min after the trial during CHO and ExCHO, although insulin was lower for ExCHO. Hyperinsulinemia during recovery in CHO and ExCHO led to an increase (P < 0.001) in PI3-kinase activity at Post-30 compared with basal, although the increase was lower (P < 0. 004) for ExCHO. Furthermore, PI3 kinase activity was suppressed (P < 0.02) immediately after exercise (Post-0) during Ex and ExCHO. Area under the insulin response curve for all trials was positively associated with PI3-kinase activity (r = 0.66, P < 0.001). Glycogen synthase activity did not increase during CHO but was increased (P < 0.05) at Post-0 and Post-30 during Ex and ExCHO. Ingestion of the drink increased (P < 0.05) carbohydrate oxidation during CHO and ExCHO, although the increase after ExCHO was lower (P < 0.05) than CHO. Carbohydrate oxidation was directly correlated with PI3-kinase activity for all trials (r = 0.63, P < 0.001). In conclusion, under resting conditions, ingestion of a carbohydrate solution led to activation of the PI3-kinase pathway and oxidation of the carbohydrate. However, when carbohydrate was ingested after intense exercise, the PI3-kinase response was attenuated and glycogen synthase activity was augmented, thus facilitating nonoxidative metabolism or storage of the carbohydrate. Activation of glycogen synthase was independent of PI3-kinase. PMID- 11007577 TI - Decrease in maximal voluntary contraction by tonic vibration applied to a single synergist muscle in humans. AB - The purpose of the study was to examine the effect of prolonged tonic vibration applied to a single synergist muscle on maximal voluntary contraction (MVC) and maximal rate of force development (dF/dt(max)). The knee extension MVC force and surface electromyogram (EMG) from the rectus femoris (RF), vastus lateralis (VL), and vastus medialis (VM) during MVC were recorded before and after vibration of RF muscle at 30 Hz for 30 min. MVC, dF/dt(max), and the integrated EMG (iEMG) of RF decreased significantly after prolonged tonic vibration in spite of no changes in iEMG of VL and VM. The present results indicate that MVC and dF/dt(max) may be influenced by the attenuated Ia afferent functions of a single synergist muscle. PMID- 11007578 TI - Hypoxia inhibits amino acid uptake in human lung fibroblasts. AB - Hypoxia and amino acid deprivation downregulate expression of extracellular matrix genes in lung fibroblasts. We examined the effect of hypoxia on amino acid uptake and protein formation in human lung fibroblasts. Low O(2) tension (0% O(2)) suppressed incorporation of [(3)H]proline into type I collagen without affecting [(35)S]methionine labeling of other proteins. Initial decreases in intracellular [(3)H]proline incorporation occurred after 2 h of exposure to 0% O(2), with maximal suppression of intracellular [(3)H]proline levels at 6 h of treatment. Hypoxia significantly inhibited the uptake of radiolabeled proline, 2 aminoisobutyric acid (AIB), and 2-(methylamino)isobutyric acid (methyl-AIB) while inducing minor decreases in leucine transport. Neither cycloheximide nor indomethacin abrogated hypoxia-related suppression of methyl-AIB uptake. Efflux studies demonstrated that hypoxia inhibited methyl-AIB transport in a bidirectional fashion. The downregulation of amino acid transport was not due to a toxic effect; function recovered on return to standard O(2) conditions. Kinetic analysis of AIB transport revealed a 10-fold increase in K(m) accompanied by a small increase in maximal transport velocity among cells exposed to 0% O(2). These data indicate that low O(2) tension regulates the system A transporter by decreasing transporter substrate affinity. PMID- 11007579 TI - Ischemic exercise and the muscle metaboreflex. AB - In exercising muscle, interstitial metabolites accumulate and stimulate muscle afferents. This evokes the muscle metaboreflex and raises arterial blood pressure (BP). In this report, we examined the effects of tension generation on muscle metabolites and BP during ischemic forearm exercise in humans. Heart rate (HR), BP, P(i), H(2)PO(4)(-), and pH ((31)P-NMR spectroscopy) data were collected in 10 normal healthy men (age 23 +/- 1 yr) during rhythmic handgrip exercise. After baseline measurements, the subjects performed rhythmic handgrip for 2 min. At 2 min, a 250-mmHg occlusion cuff was inflated, and ischemic handgrip exercise was continued until near fatigue (Borg 19). Measurements were continued for an additional 30 s of ischemia. This protocol was performed at 15, 30, 45, and 60% of the subjects' maximum voluntary contraction (MVC) in random order. As tension increased, the time to fatigue decreased. In addition, mean arterial pressure and HR were higher at 60% MVC than at any of the other lower tensions. The NMR data showed significantly greater increases in H(2)PO(4)(-), P(i), and H(+) at 60% than at 15 and 30% MVC. Therefore, despite the subjects working to the same perceived effort level, a greater reflex response (represented by BP and HR data) was elicited at 60% MVC than at any of the other ischemic tensions. These data are consistent with the hypothesis that, as tension increases, factors aside from insufficient blood flow contribute to the work effect on muscle metabolites and the magnitude of the reflex response. PMID- 11007580 TI - Diaspirin cross-linked hemoglobin improves oxygen extraction capabilities in endotoxic shock. AB - We studied the effects of diaspirin cross-linked hemoglobin (DCLHb), a cell-free hemoglobin derived from human erythrocytes, on blood flow distribution and tissue oxygen extraction capabilities in endotoxic shock. Eighteen pentobarbital sodium anesthetized, mechanically ventilated dogs received 2 mg/kg of E. coli endotoxin, followed by saline resuscitation to restore cardiac filling pressures to baseline levels. The animals were randomly divided into three groups: six served as control, six received DCLHb at a dose of 500 mg/kg (group 1) and six DCLHb at a dose of 1,000 mg/kg (group 2). Cardiac tamponade was then induced by saline injection in the pericardial sac to progressively reduce cardiac index and thereby allow study of tissue oxygen extraction capabilities. DCLHb had a dose dependent vasopressor effect but did not significantly alter cardiac index or regional blood flow. During cardiac tamponade, critical oxygen delivery was 12.8 +/- 0.7 ml. kg(-1). min(-1) in the control group, but 8.6 +/- 0.9 and 8.2 +/- 0.7 ml. kg(-1). min(-1) in groups 1 and 2, respectively (both P < 0.05 vs. control group). The critical oxygen extraction ratio was 39.1 +/- 3.1% in the control group but 58.7 +/- 12.8% and 60.2 +/- 9.0% in groups 1 and 2, respectively. We conclude that DCLHb can improve whole body oxygen extraction capabilities during endotoxic shock in dogs. PMID- 11007581 TI - Echocardiographic criteria for detection of postinfarction congestive heart failure in rats. AB - We evaluated postinfarction myocardial function in rats and determined echocardiographic criteria for congestive heart failure (CHF) using high performance echocardiography. Extensive myocardial infarction (MI) was induced in rats by left coronary occlusion. Sham-operated animals served as controls. Five weeks later, high-frame rate ( approximately 200 Hz), fully digitized, shallow focus (10-25 mm), two-dimensional, M-mode and Doppler echocardiography was performed. A J-tree cluster analysis was performed using parameters indicative of CHF. Reproducibility was examined. The cluster analysis joined the animals into one Sham and two MI clusters. One of the MI clusters had clinical characteristics of CHF and elevated left ventricular end diastolic pressure. Among the echocardiographic variables, only posterior wall shortening velocity separated the failing and nonfailing MI clusters. We conclude that, by high frame rate echocardiography, it is possible to obtain high- quality recordings in rats. It is feasible to distinguish MI rats with CHF due to myocardial dysfunction from those without failure and to perform longitudinal studies on myocardial function. PMID- 11007583 TI - Collagen gene expression in rat left ventricle: interactive effect of age and exercise training. AB - Whether or not exercise training of sufficient intensity and duration to produce left ventricle (LV) hypertrophy also regulates deposition of interstitial collagen and cross-linking at the pretranslational level is unknown. Therefore, the effects of exercise training on gene expression for the two principal fibrillar collagens in LV, types I and III, were assessed in young adult (5 mo), middle-aged (15 mo), and old (26 mo) rats. We also evaluated the potential interaction of changes in mRNA for these procollagens with alterations in LV extracellular matrix characteristics by simultaneously measuring collagen concentration (hydroxyproline) and extent of mature collagen cross-linking (hydroxylysylpyridinoline, HP). Ten weeks of treadmill running resulted in LV hypertrophy and an increased maximal oxygen uptake in all three age groups of trained rats compared with sedentary controls. Percent collagen in rat LV almost doubled (P < 0.0001) from 5 to 26 mo of age, an increase unaffected by exercise training. With aging, a significant decline in expression of mRNAs for both collagen type I (P < 0.005) and type III (P < 0.001) was observed in LV free wall (LVF) but not septum (LVS). Training prevented this decline in LVF mRNAs for the two principal fibrillar collagens in middle-aged rats whereas it attenuated the decline in senescent animals. HP concentration increased significantly with aging in both LVF (P < 0.005) and LVS (P < 0.01). Training modulated this effect, but again only in LVF, so that HP was significantly lower (P < 0.05) in this region of the LV in old trained rats compared with sedentary counterparts. We conclude that exercise training modulates the effects of aging on collagen gene mRNAs and HP cross-linking regionally within the LV. PMID- 11007582 TI - Effects of aortic nerve on hemodynamic response to obstructive apnea in sedated pigs. AB - In this study we test the hypothesis that aortic nerve traffic is responsible for the pressor response to periodic apneas. In nine intubated, sedated chronically instrumented pigs, periodic obstructive apneas were caused by occlusion of the endotracheal tube for 30 s, followed by spontaneous breathing for 30 s. This was done under control (C) conditions, after section of the aortic nerve (ANS), and after bilateral cervical vagotomy (Vagot). Blood-gas tensions and airway pressure changed similarly under all conditions: PO(2) decreased to 50-60 Torr, PCO(2) increased to approximately 55 Torr, and airway pressure decreased by 40-50 mmHg during apnea. With C, mean arterial pressure (MAP) increased from 111 +/- 4 mmHg at baseline to 120 +/- 5 mmHg at late apnea (P < 0.01). After ANS and Vagot, there was no change in MAP with apneas compared with baseline. Relative to baseline, cardiac output and stroke volume decreased with C but not with ANS or Vagot during apneas. Increased MAP was due to increased systemic vascular resistance. Heart rate behaved similarly with C and ANS, being greater at early interapnea than late apnea. With Vagot, heart rate increased throughout the apnea interapnea cycle relative to baseline. We conclude that, in sedated pigs, aortic nerve traffic mediates the increase in MAP and systemic vascular resistance observed during periodic apneas. Increase in MAP is responsible for decreased cardiac output and stroke volume. Additional vagal reflexes, most likely parasympathetic efferents, are responsible for interacting with sympathetic excitatory influences in modulating heart rate. PMID- 11007584 TI - Synergist muscle ablation and recovery from nerve-repair grafting: contractile and metabolic function. AB - After nerve-repair grafting of medial gastrocnemius muscle, there is incomplete recovery of specific force and sustainable power, perhaps due to overcompensation by synergistic muscles. We hypothesized that increased workload due to synergist ablation would enhance graft recovery. Contractile and metabolic properties of control and nerve-repair grafted muscles, with and without synergist ablation, were determined after 120 days recovery. Specific force (N/cm(2)) and normalized power (W/kg) were less in the experimental groups compared with controls. Sustained power (W/kg) in the synergist-ablated nerve-repair grafted muscle was higher than nerve-repair grafted muscle, returning to control values. GLUT-4 protein was higher and glycogen content was diminished in both synergist-ablated groups. In summary, synergist ablation did not enhance the recovery of specific force or normalized power, but sustained power did recover, suggesting that metabolic and not mechanical parameters were responsible for this recovery. The enhanced endurance after synergist ablation was accompanied by increased GLUT-4 protein, suggesting a role for increased uptake of circulating glucose during contraction. PMID- 11007585 TI - Hyperbaric oxygen improves contractile function of regenerating rat skeletal muscle after myotoxic injury. AB - There is growing interest in hyperbaric oxygen (HBO) as an adjunctive treatment for muscle injuries. This experiment tested the hypothesis that periodic inhalation of HBO hastens the functional recovery and myofiber regeneration of skeletal muscle after myotoxic injury. Injection of the rat extensor digitorum longus (EDL) muscle with bupivacaine hydrochloride causes muscle degeneration. After injection, rats breathed air with or without periodic HBO [100% O(2) at either 2 or 3 atmospheres absolute (ATA)]. In vitro maximum isometric tetanic force of injured EDL muscles and regenerating myofiber size were unchanged between 2 ATA HBO-treated and untreated rats at 14 days postinjury but were approximately 11 and approximately 19% greater, respectively, in HBO-treated rats at 25 days postinjury. Maximum isometric tetanic force of injured muscles was approximately 27% greater, and regenerating myofibers were approximately 41% larger, in 3 ATA HBO-treated rats compared with untreated rats at 14 days postinjury. These findings demonstrate that periodic HBO inhalation increases maximum force-producing capacity and enhances myofiber growth in regenerating skeletal muscle after myotoxic injury with greater effect at 3 than at 2 ATA. PMID- 11007586 TI - Hindlimb unweighting alters endothelium-dependent vasodilation and ecNOS expression in soleus arterioles. AB - The purpose of this study was to test the hypothesis that endothelium-dependent dilation is impaired in soleus resistance arteries from hindlimb-unweighted (HLU) rats. Male Sprague-Dawley rats (300-350 g) were exposed to HLU (n = 14) or weight bearing control (Con, n = 14) conditions for 14 days. After the 14-day treatment period, soleus first-order (1A) arterioles were isolated and cannulated with micropipettes to assess vasodilator responses to an endothelium-dependent dilator, ACh (10(-9)-10(-4) M), and an endothelium-independent dilator, sodium nitroprusside (SNP, 10(-9)-10(-4) M). Arterioles from HLU rats were smaller than Con arterioles (maximal passive diameter = 140 +/- 4 and 121 +/- 4 microm in Con and HLU, respectively) but developed similar spontaneous myogenic tone (43 +/- 3 and 45 +/- 3% in Con and HLU, respectively). Arteries from Con and HLU rats dilated in response to increasing doses of ACh, but dilation was impaired in arterioles from HLU rats (P = 0.03), as was maximal dilation to ACh (85 +/- 4 and 65 +/- 4% possible dilation in Con and HLU, respectively). Inhibition of nitric oxide (NO) synthase (NOS) with N(omega)-nitro-L-arginine (300 microM) reduced ACh dilation by approximately 40% in arterioles from Con rats and eliminated dilation in arterioles from HLU rats. The cyclooxygenase inhibitor indomethacin (50 microM) did not significantly alter dilation to ACh in either group. Treatment with N(omega)-nitro-L-arginine + indomethacin eliminated all ACh dilation in Con and HLU rats. Dilation to sodium nitroprusside was not different between groups (P = 0.98). To determine whether HLU decreased expression of endothelial cell NOS (ecNOS), mRNA and protein levels were measured in single arterioles with RT-PCR and immunoblot analysis. The ecNOS mRNA and protein expression was significantly lower in arterioles from HLU rats than in Con arterioles (20 and 65%, respectively). Collectively, these data indicate that HLU impairs ACh dilation in soleus 1A arterioles, in part because of alterations in the NO pathway. PMID- 11007587 TI - Effects of 2-G exposure on temperature regulation, circadian rhythms, and adiposity in UCP2/3 transgenic mice. AB - Altered ambient force environments affect energy expenditure via changes in thermoregulation, metabolism, and body composition. Uncoupling proteins (UCPs) have been implicated as potential enhancers of energy expenditure and may participate in some of the adaptations to a hyperdynamic environment. To test this hypothesis, this study examined the homeostatic and circadian profiles of body temperature (T(b)) and activity and adiposity in wild-type and UCP2/3 transgenic mice exposed to 1 and 2 G. There were no significant differences between the groups in the means, amplitudes, or phases of T(b) and activity rhythms at either the 1- or 2-G level. Percent body fat was significantly lower in transgenic (5.2 +/- 0. 2%) relative to the wild-type mice (6.2 +/- 0.1%) after 2-G exposure; mass-adjusted mesenteric and epididymal fat pads in transgenic mice were also significantly lower (P < 0.05). The data suggest that 1) the actions of two UCPs (UCP2 and UCP3) do not contribute to an altered energy balance at 2 G, although 2) UCP2 and UCP3 do contribute to the utilization of lipids as a fuel substrate at 2 G. PMID- 11007588 TI - Exercise elevates plasma levels but not gene expression of IL-1beta, IL-6, and TNF-alpha in blood mononuclear cells. AB - Physical activity induces a subclinical inflammatory response, mediated in part by leukocytes, and manifested by elevated concentrations of circulating proinflammatory cytokines, including interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha). However, the source of the cytokines that appear during exercise remains unknown. In this study, we examined exercise induced changes in plasma cytokine concentrations and their corresponding mRNA expression in peripheral blood mononuclear cells. Ten healthy [peak oxygen uptake = 48.8 +/- 6.5 (SD) ml. kg(-1). min(-1)] but untrained men [age = 25 +/- 5 (SD) yr] undertook 3 h of exercise (cycling and inclined walking) at 60-65% peak oxygen uptake. Circulating leukocyte subset counts were elevated during and 2 h postexercise but returned to normal within 24 h. Plasma concentrations of IL 1beta, IL-6, and TNF-alpha peaked at the end of exercise and remained elevated at 2 h (IL-6) and up to 24 h (IL-1beta and TNF-alpha) postexercise. Cytokine gene expression in circulating mononuclear cells was measured by using the reverse transcriptase-polymerase chain reaction; mRNA accumulation did not change with exercise. In conclusion, mRNA accumulation of IL-1beta, IL-6, and TNF-alpha in circulating mononuclear cells is not affected by 3 h of moderate endurance exercise and does not seem to account for the observed increases in plasma cytokines. PMID- 11007589 TI - Vascular perturbations in the chronic orthostatic intolerance of the postural orthostatic tachycardia syndrome. AB - Chronic orthostatic intolerance is often related to the postural orthostatic tachycardia syndrome (POTS). POTS is characterized by upright tachycardia. Understanding of its pathophysiology remains incomplete, but edema and acrocyanosis of the lower extremities occur frequently. To determine how arterial and venous vascular properties account for these findings, we compared 13 patients aged 13-18 yr with 10 normal controls. Heart rate and blood pressure were continuously recorded, and strain-gauge plethysmography was used to measure forearm and calf blood flow, venous compliance, and microvascular filtration while the subject was supine and to measure calf blood flow and calf size change during head-up tilt. Resting venous pressure was higher in POTS compared with control (16 vs. 10 mmHg), which gave the appearance of decreased compliance in these patients. The threshold for edema formation decreased in POTS patients compared with controls (8.3 vs. 16.3 mmHg). With tilt, early calf blood flow increased in POTS patients (from 3.4 +/- 0.9 to 12.6 +/- 2.3 ml. 100 ml(-1). min( 1)) but did not increase in controls. Calf volume increased twice as much in POTS patients compared with controls over a shorter time of orthostasis. The data suggest that resting venous pressure is higher and the threshold for edema is lower in POTS patients compared with controls. Such findings make the POTS patients particularly vulnerable for edema fluid collection. This may signify a redistribution of blood to the lower extremities even while supine, accounting for tachycardia through vagal withdrawal. PMID- 11007590 TI - Unilateral lung edema: effects on pulmonary gas exchange, hemodynamics, and pulmonary perfusion distribution. AB - Two types of unilateral lung edema in sheep were characterized regarding their effects on pulmonary gas exchange, hemodynamics, and distribution of pulmonary perfusion. One edema type was induced with aerosolized HCl (0.15 M, pH 1.0) and the other with NaCl (0.15 M, pH 7.4). Both aerosols were nebulized continuously for 4 h into left lungs. In HCl-treated animals, pulmonary gas exchange deteriorated [from a partial arterial O(2) pressure-to-inspired O(2) fraction ratio (Pa(O(2))/FI(O(2))) of 254 at baseline to 187 after 4 h HCl]. In addition, pulmonary artery pressure and total pulmonary vascular resistance increased (from 16 to 19 mmHg and from 133 to 154 dyn. s. cm(-5), respectively). In NaCl-treated animals, only the central venous pressure significantly increased (from 7 to 9 mmHg). Distribution of pulmonary perfusion (measured with fluorescent microspheres) changed differently in both groups. After HCl application, 6% more blood flow was directed to the treated lung, whereas, after NaCl, 5% more blood flow was directed to the untreated lung. HCl and NaCl treatment both induce an equivalent lung edema, but only HCl treatment is associated with gas exchange alteration and tissue damage. Redistribution of pulmonary perfusion maintains gas exchange during NaCl treatment and decreases it during HCl inhalation. PMID- 11007591 TI - Scientific approach to the 1-h cycling world record: a case study. AB - The purpose of this study was to describe the physiological and aerodynamic characteristics and the preparation for a successful attempt to break the 1-h cycling world record. An elite professional road cyclist (30 yr, 188 cm, 81 kg) performed an incremental laboratory test to assess maximal power output (W(max)) and power output (W(OBLA)), estimated speed (V(OBLA)), and heart rate (HR(OBLA)) at the onset of blood lactate accumulation (OBLA). He also completed an incremental velodrome (cycling track) test (VT1), during which V(OBLAVT1) and HR(OBLAVT1) were measured and W(OBLAVT1) was estimated. W(max) was 572 W, W(OBLA) 505 W, V(OBLA) 52.88 km/h, and HR(OBLA) 183 beats/min. V(OBLAVT1), HR(OBLAVT1), and W(OBLAVT1) were 52.7 km/h, 180 beats/min, and 500.6 W, respectively. Drag coefficient and shape coefficient, measured in a wind tunnel, were 0. 244 and 0.65 m(2), respectively. The cyclist set a world record of 53,040 m, with an estimated average power output of 509.5 W. Based on direct laboratory data of the power vs. oxygen uptake relationship for this cyclist, this is slightly higher than the 497. 25 W corresponding to his oxygen uptake at OBLA (5.65 l/min). In conclusion, 1) the 1-h cycling world record is the result of the interaction between physiological and aerodynamic characteristics; and 2) performance in this event can be predicted using mathematical models that integrate the principal performance-determining variables. PMID- 11007592 TI - 5-Hydroxytryptophan-induced respiratory recovery after cervical spinal cord hemisection in rats. AB - The present study investigates the role of serotonin in respiratory recovery after spinal cord injury. Experiments were conducted on C(2) spinal cord hemisected, anesthetized, vagotomized, paralyzed, and artificially ventilated rats in which end-tidal CO(2) was monitored and maintained. Before drug administration, the phrenic nerve ipsilateral to hemisection showed no respiratory-related activity due to the disruption of the descending bulbospinal respiratory pathways by spinal cord hemisection. 5-Hydroxytryptophan (5-HTP), a serotonin precursor, was administrated intravenously. 5-HTP induced time- and dose-dependent increases in respiratory recovery in the phrenic nerve ipsilateral to hemisection. Although the 5-HTP-induced recovery was initially accompanied by an increase in activity in the contralateral phrenic nerve, suggesting an increase in descending respiratory drive, the recovery persisted well after activity in the contralateral nerve returned to predrug levels. 5-HTP-induced effects were reversed by a serotonin receptor antagonist, methysergide. Because experiments were conducted on animals subjected to C(2) spinal cord hemisection, the recovery was most likely mediated by the activation of a latent respiratory pathway spared by the spinal cord injury. The results suggest that serotonin is an important neuromodulator in the unmasking of the latent respiratory pathway after spinal cord injury. In addition, the results also suggest that the maintenance of 5-HTP-induced respiratory recovery may not require a continuous enhancement of central respiratory drive. PMID- 11007593 TI - Alveolar epithelial integrity in athletes with exercise-induced hypoxemia. AB - The effect of incremental exercise to exhaustion on the change in pulmonary clearance rate (k) of aerosolized (99m)Tc-labeled diethylenetriaminepentaacetic acid ((99m)Tc-DTPA) and the relationship between k and arterial PO(2) (Pa(O(2))) during heavy work were investigated. Ten male cyclists (age = 25 +/- 2 yr, height = 180.9 +/- 4.0 cm, mass = 80.1 +/- 9.5 kg, maximal O(2) uptake = 5. 25 +/- 0.35 l/min, mean +/- SD) completed a pulmonary clearance test shortly (39 +/- 8 min) after a maximal O(2) uptake test. Resting pulmonary clearance was completed >/=24 h before or after the exercise test. Arterial blood was sampled at rest and at 1 min intervals during exercise. Minimum Pa(O(2)) values and maximum alveolar arterial PO(2) difference ranged from 73 to 92 Torr and from 30 to 55 Torr, respectively. No significant difference between resting k and postexercise k for the total lung (0.55 +/- 0.20 vs. 0. 57 +/- 0.17 %/min, P > 0.05) was observed. Pearson product-moment correlation indicated no significant linear relationship between change in k for the total lung and minimum Pa(O(2)) (r = -0.26, P > 0.05). These results indicate that, averaged over subjects, pulmonary clearance of (99m)Tc-DTPA after incremental maximal exercise to exhaustion in highly trained male cyclists is unchanged, although the sampling time may have eliminated a transient effect. Lack of a linear relationship between k and minimum Pa(O(2)) during exercise suggests that exercise-induced hypoxemia occurs despite maintenance of alveolar epithelial integrity. PMID- 11007594 TI - pH(i) responses to osmotic cell shrinkage in the presence of open-system buffers. AB - Changes in plasma volume in vivo cause rapid changes in extracellular pH by altering the plasma bicarbonate concentration at a constant Pco(2) (Garella S, Chang BS, and Kahn SI. Kidney Int 8: 279, 1975). Few studies have examined the possibility that changes in cell volume produce comparable changes in intracellular pH (pH(i)). In the present study, alveolar macrophages were exposed to hyperosmotic medium in the absence or presence of the open-system buffers CO(2)-HCO(3)(-), propionic acid-propionate, or NH(3)-NH(4)(+). In the absence of open-system buffers, exposure to twice-normal osmolarity (2T) produced a slow cellular alkalinization [change in pH(i) (DeltapH(i)) approximately 0.38; exponential time constant (tau) approximately 120 s]. In the presence of 5% CO(2), 2T caused a biphasic pH(i) response: a rapid increase (DeltapH(i) approximately 0.10, tau approximately 15 s) followed by a slower pH(i) increase. Identical rapid pH(i) increases were produced by 2T in the presence of propionic acid (20 mM). Conversely, 2T caused a rapid pH(i) decrease (DeltapH(i) approximately -0.21, tau approximately 10 s) in the presence of NH(3) (20 mM). Thus osmotic cell shrinkage caused rapid pH(i) changes of opposite direction in the presence of a weak acid buffer (contraction alkalosis with CO(2) or propionic acid) vs. a weak base buffer (contraction acidosis with NH(3)). Graded DeltapH(i) were produced by varying extracellular osmolarity in the presence of open-system buffers; osmolarity increases of as little as 5-10% produced significant DeltapH(i). The rapid pH(i) responses to 2T were insensitive to inhibitors of membrane H(+) transport (ethylisopropylamiloride and bafilomycin A(1)). The results are consistent with shrinkage-induced disequilibria in the total cellular buffer system (i.e., intrinsic buffers plus added weak acid-base buffer). PMID- 11007595 TI - A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation. AB - Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100% O(2) at 4 degrees C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37 degrees C) DMEM plus 10% fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia reperfusion injury at the cellular and molecular level. PMID- 11007596 TI - Systemic hypoxia increases leukocyte emigration and vascular permeability in conscious rats. AB - We recently observed that acute systemic hypoxia produces rapid increases in leukocyte adherence in the mesenteric microcirculation of the anesthetized rat Wood JG, Johnson JS, Mattioli LF, and Gonzalez NC. J Appl Physiol 87: 1734-1740, 1999; Wood JG, Mattioli LF, and Gonzalez NC. J Appl Physiol 87: 873-881, 1999. Hypoxia-induced leukocyte adherence is associated with an increase in reactive oxygen species (ROS) generation and is attenuated by antioxidants or interventions that increase tissue levels of nitric oxide (NO). These results suggest that the acute effects of hypoxia on leukocyte-endothelial interactions are caused by a change in the ROS-NO balance. The present experiments were designed to extend our observations of the initial microcirculatory response to hypoxia; specifically, we wanted to determine whether the response to systemic hypoxia involves increased microvascular permeability and leukocyte emigration and whether ROS generation and decreased NO levels contribute to these responses. At this time, there is conflicting evidence, from in vitro studies, regarding the effect of hypoxia on these indexes of vascular function. Our studies were carried out in the physiological setting of the conscious animal, in which a prolonged hypoxic exposure is possible without the adverse effects that may develop under anesthesia. The central observation of these studies is that conscious animals exposed for 4 h to environmental hypoxia show increased microvascular permeability and emigration of leukocytes into the extravascular space of the mesenteric circulation. Furthermore, these events are dependent on increased ROS generation and, possibly, a subsequent decrease in tissue NO levels during systemic hypoxia. Our results show that systemic hypoxia profoundly affects vascular endothelial function through changes in the ROS-NO balance in the conscious animal. PMID- 11007597 TI - Electrical admittance for filling of the heart during lower body negative pressure in humans. AB - To evaluate whether electrical admittance of intracellular water is applicable for monitoring filling of the heart, we determined the difference in intracellular water in the thorax (Thorax(ICW)), measured as the reciprocal value of the electrical impedance for the thorax at 1.5 and 100 kHz during lower body negative pressure (LBNP) in humans. Changes in Thorax(ICW) were compared with positron emission tomography-determined C(15)O-labeled erythrocytes over the heart. During -40 mmHg LBNP, the blood volume of the heart decreased by 21 +/- 3% as the erythrocyte volume was reduced by 20 +/- 2% and the plasma volume declined by 26 +/- 2% (P < 0.01; n = 8). Over the heart region, LBNP was also associated with a decrease in the technetium-labeled erythrocyte activity by 26 +/- 4% and, conversely, an increase over the lower leg by 92 +/- 5% (P < 0.01; n = 6). For 15 subjects, LBNP increased thoracic impedance by 3.3 +/- 0.3 Omega (1.5 kHz) and 3.0 +/- 0.4 Omega (100 kHz), whereas leg impedance decreased by 9.0 +/- 3.3 Omega (1.5 kHz) and 6.1 +/- 3 Omega (100 kHz; P < 0.01). Thorax(ICW) was reduced by 7.1 +/- 1.9 S. 10(-4) (P < 0.01) and intracellular water in the leg tended to increase (from 37.8 +/- 4.6 to 40.9 +/- 5.0 S. 10(-4); P = 0.08). The correlation between Thorax(ICW) and heart erythrocyte volume was 0.84 (P < 0.05). The results suggest that thoracic electrical admittance of intracellular water can be applied to evaluate changes in blood volume of the heart during LBNP in humans. PMID- 11007598 TI - Description and validation of a novel liquid metal-free device for venous congestion plethysmography. AB - We present a newly developed electromechanical sensor with automated calibration for strain-gauge plethysmography (filtrass) and compare it to a conventional mercury-in-Silastic strain-gauge plethysmograph (MSG). Fluid filtration capacity (K(f)) and isovolumetric venous pressure (Piv) of the limb were assessed noninvasively with both devices in 29 healthy volunteers. We found significantly higher K(f) and Piv values with MSG [4.6 +/- 2.0 x 10(-3) ml. min(-1). mmHg(-1). 100 ml tissue(-1) (K(f) units; K(f)U) and 21.2 +/- 8.1 mmHg for Pvi], than with filtrass, giving values of 3.1 +/- 0.8 K(f)U and 15.1 +/- 7.1 mmHg. Because K(f) and Piv are profoundly influenced by the calibration, we investigated the quality of the calibration signal and its impact on the obtained values. We could show that the reproducibility of repeated calibrations was higher with filtrass (58% lower mean +/- SD). The data were grouped according to the quality of calibration, and we found no significant difference in K(f) and Piv between filtrass (3.0 +/- 0.7 K(f)U and 15.9 +/- 6.9 mmHg, respectively) and MSG with good calibration signal (3.3 +/- 0. 8 K(f)U and 18.6 +/- 7.1 mmHg, respectively; no significant difference). However, we obtained significantly higher MSG values (5. 6 +/- 2.0 K(f)U and 23.1 +/- 8.4 mmHg, respectively; P < 0.001) in the group with a bad calibration signal. We suggest that the filtrass sensor, which performs an automatic, standardized calibration procedure and shows a linear signal response to stretch, gives highly reproducible and reliable results and thus is more suitable for routine application. PMID- 11007599 TI - Effect of transpulmonary pressure on airway diameter and responsiveness of immature and mature rabbits. AB - We previously demonstrated that airway responsiveness is greater in immature than in mature rabbits; however, it is not known whether there are maturational differences in the effect of transpulmonary pressure (Ptp) on airway size and airway responsiveness. The relationship between Ptp and airway diameter was assessed in excised lungs insufflated with tantalum powder. Diameters of comparable intraparenchymal airway segments were measured from radiographs obtained at Ptp between 0 and 20 cmH(2)O. At Ptp > 8 cmH(2)O, the diameters were near maximal in both groups. With diameter normalized to its maximal value, changing Ptp between 8 and 0 cmH(2)O resulted in a greater decline of airway caliber in immature than mature airways. The increases in lung resistance (RL) in vivo at Ptp of 8, 5, and 2 cmH(2)O were measured during challenge with intravenous methacholine (MCh: 0.001-0.5 mg/kg). At Ptp of 8 cmH(2)O, both groups had very small responses to MCh and the maximal fold increases in RL did not differ (1.93 +/- 0.29 vs. 2.23 +/- 0.19). At Ptp of 5 and 2 cmH(2)O, the fold increases in RL were greater for immature than mature animals (13.19 +/- 1.81 vs. 3.89 +/- 0.37) and (17.74 +/- 2.15 vs. 4.6 +/- 0.52), respectively. We conclude that immature rabbits have greater airway distensibility and this difference may contribute to greater airway narrowing in immature compared with mature rabbits. PMID- 11007600 TI - Influence of lung volume on pulmonary microvascular pressure-volume characteristics. AB - The pressure-volume (P-V) characteristics of the lung microcirculation are important determinants of the pattern of pulmonary perfusion and of red and white cell transit times. Using diffuse light scattering, we measured capillary P-V loops in seven excised perfused dog lobes at four lung volumes, from functional residual capacity (FRC) to total lung capacity (TLC), over a wide range of vascular transmural pressures (Ptm). At Ptm 5 cmH(2)O, specific compliance of the microvasculature was 8.6%/cmH(2)O near FRC, decreasing to 2.7%/cmH(2)O as lung volume increased to TLC. At low lung volumes, the vasculature showed signs of strain stiffening (specific compliance fell as Ptm rose), but stiffening decreased as lung volume increased and was essentially absent at TLC. The P-V loops were smooth without sharp transitions, consistent with vascular distension as the primary mode of changes in vascular volume with changes in Ptm. Hysteresis was small (0.013) at all lung volumes, suggesting that, although surface tension may set basal capillary shape, it does not strongly affect capillary compliance. PMID- 11007601 TI - Influence of microgravity on crystal formation in biomineralization. AB - Biomineralized tissues are widespread in animals. They are essential elements in skeletons and in statocysts. The function of both can only be understood with respect to gravitational force, which has always been present. Therefore, it is not astonishing to identify microgravity as a factor influencing biomineralization, normally resulting in the reduction of biomineralized materials. All known biominerals are composite materials, in which the organic matrix and the inorganic materials, organized in crystals, interact. If, during remodeling and turnover processes under microgravity, a defective organization of these crystals occurs, a reduction in biomineralized materials could be the result. To understand the influence of microgravity on the formation of biocrystals, we studied the shell-building process of the snail Biomphalaria glabrata as a model system. We show that, under microgravity (space shuttle flights STS-89 and STS-90), shell material is built in a regular way in both adult snails and snail embryos during the beginning of shell development. Microgravity does not influence crystal formation. Because gravity has constantly influenced evolution, the organization of biominerals with densities near 3 must have gained independence from gravitational forces, possibly early in evolution. PMID- 11007602 TI - Effects of voluntary activity and genetic selection on muscle metabolic capacities in house mice Mus domesticus. AB - Selective breeding is an important tool in behavioral genetics and evolutionary physiology, but it has rarely been applied to the study of exercise physiology. We are using artificial selection for increased wheel-running behavior to study the correlated evolution of locomotor activity and physiological determinants of exercise capacity in house mice. We studied enzyme activities and their response to voluntary wheel running in mixed hindlimb muscles of mice from generation 14, at which time individuals from selected lines ran more than twice as many revolutions per day as those from control (unselected) lines. Beginning at weaning and for 8 wk, we housed mice from each of four replicate selected lines and four replicate control lines with access to wheels that were free to rotate (wheel-access group) or locked (sedentary group). Among sedentary animals, mice from selected lines did not exhibit a general increase in aerobic capacities: no mitochondrial [except pyruvate dehydrogenase (PDH)] or glycolytic enzyme activity was significantly (P < 0.05) higher than in control mice. Sedentary mice from the selected lines exhibited a trend for higher muscle aerobic capacities, as indicated by higher levels of mitochondrial (cytochrome-c oxidase, carnitine palmitoyltransferase, citrate synthase, and PDH) and glycolytic (hexokinase and phosphofructokinase) enzymes, with concomitant lower anaerobic capacities, as indicated by lactate dehydrogenase (especially in male mice). Consistent with previous studies of endurance training in rats via voluntary wheel running or forced treadmill exercise, cytochrome-c oxidase, citrate synthase, and carnitine palmitoyltransferase activity increased in the wheel-access groups for both genders; hexokinase also increased in both genders. Some enzymes showed gender specific responses: PDH and lactate dehydrogenase increased in wheel-access male but not female mice, and glycogen phosphorylase decreased in female but not in male mice. Two-way analysis of covariance revealed significant interactions between line type and activity group; for several enzymes, activities showed greater changes in mice from selected lines, presumably because such mice ran more revolutions per day and at greater velocities. Thus genetic selection for increased voluntary wheel running did not reduce the capability of muscle aerobic capacity to respond to training. PMID- 11007603 TI - Biology lessons from oscillatory cell mechanics. PMID- 11007604 TI - Mechanical properties of cultured human airway smooth muscle cells from 0.05 to 0.4 Hz. AB - We investigated the rheological properties of living human airway smooth muscle cells in culture and monitored the changes in rheological properties induced by exogenous stimuli. We oscillated small magnetic microbeads bound specifically to integrin receptors and computed the storage modulus (G') and loss modulus (G") from the applied torque and the resulting rotational motion of the beads as determined from their remanent magnetic field. Under baseline conditions, G' increased weakly with frequency, whereas G" was independent of the frequency. The cell was predominantly elastic, with the ratio of G" to G' (defined as eta) being approximately 0. 35 at all frequencies. G' and G" increased together after contractile activation and decreased together after deactivation, whereas eta remained unaltered in each case. Thus elastic and dissipative stresses were coupled during changes in contractile activation. G' and G" decreased with disruption of the actin fibers by cytochalasin D, but eta increased. These results imply that the mechanisms for frictional energy loss and elastic energy storage in the living cell are coupled and reside within the cytoskeleton. PMID- 11007605 TI - Brief commentary on coronary wave-intensity analysis. PMID- 11007606 TI - Wave-intensity analysis: a new approach to coronary hemodynamics. AB - In 10 anesthetized dogs, we measured high-fidelity left circumflex coronary (P(LCx)), aortic (P(Ao)), and left ventricular (P(LV)) pressures and left circumflex velocity (U(LCx); Doppler) and used wave-intensity analysis (WIA) to identify the determinants of P(LCx) and U(LCx). Dogs were paced from the right atrium (control 1) or right ventricle by use of single (control 2) and then paired pacing to evaluate the effects of left ventricular contraction on P(LCx) and U(LCx). During left ventricular isovolumic contraction, P(LCx) exceeded P(Ao), paired pacing increasing the difference. Paired pacing increased DeltaP(X) (the P(LCx)-P(Ao) difference at the P(Ao)-P(LV) crossover) and average dP(LCx)/dt (P < 0.0001 for both). During this time, WIA identified a backward-going compression wave (BCW) that increased P(LCx) and decreased U(LCx); the BCW increased during paired pacing (P < 0.0001). After the aortic valve opened, the increase in P(Ao) caused a forward-going compression wave that, when it exceeded the BCW, caused U(LCx) to increase, despite P(LV) and (presumably) elastance continuing to increase. Thus WIA identifies the contributions of upstream (aortic) and downstream (microcirculatory) effects on P(LCx) and U(LCx). PMID- 11007608 TI - commentary PMID- 11007607 TI - Invited review: mechanisms of ventilator-induced lung injury: a perspective. AB - Despite advances in critical care, the mortality rate in patients with acute lung injury remains high. Furthermore, most patients who die do so from multisystem organ failure. It has been postulated that ventilator-induced lung injury plays a key role in determining the negative clinical outcome of patients exposed to mechanical ventilation. How mechanical ventilation exerts its detrimental effect is as of yet unknown, but it appears that overdistension of lung units or shear forces generated during repetitive opening and closing of atelectatic lung units exacerbates, or even initiates, significant lung injury and inflammation. The term "biotrauma" has recently been elaborated to describe the process by which stress produced by mechanical ventilation leads to the upregulation of an inflammatory response. For mechanical ventilation to exert its deleterious effect, cells are required to sense mechanical forces and activate intracellular signaling pathways able to communicate the information to its interior. This information must then be integrated in the nucleus, and an appropriate response must be generated to implement and/or modulate its response and that of neighboring cells. In this review, we present a perspective on ventilator-induced lung injury with a focus on mechanisms and clinical implications. We highlight some of the most recent findings, which we believe contribute to the generation and propagation of ventilator-induced lung injury, placing a special emphasis on their implication for future research and clinical therapies. PMID- 11007609 TI - Invited review: effects of flow on vascular endothelial intracellular calcium signaling of rat aortas ex vivo. AB - To study the effects of flow on in situ endothelial intracellular calcium concentration ([Ca(2+)](i)) signaling, rat aortic rings were loaded with fura 2, mounted on a tissue flow chamber, and divided into control and flow-pretreated groups. The latter was perfused with buffer at a shear stress of 50 dyns/cm(2) for 1 h. Endothelial [Ca(2+)](i) responses to ACh or shear stresses were determined by ratio image analysis. Moreover, ACh-induced [Ca(2+)](i) elevation responses were measured in a calcium-free buffer, or in the presence of SKF 96365, to elucidate the role of calcium influx in the flow effects. Our results showed that 1) ACh increased endothelial [Ca(2+)](i) in a dose-dependent manner, and these responses were incremented by flow-pretreatment; 2) the differences in ACh-induced [Ca(2+)](i) elevation between control and flow-pretreated groups were abolished by SKF-96365 or by Ca(2+)-free buffer; and 3) in the presence of 10(-5) M ATP, shear stress induced dose-dependent [Ca(2+)](i) elevation responses that were not altered by flow-pretreatment. In conclusion, flow-pretreatment augments the ACh-induced endothelial calcium influx in rat aortas ex vivo. PMID- 11007610 TI - Opposing views on tensegrity as a structural framework for understanding cell mechanics. PMID- 11007611 TI - The foreign body reaction to a biodegradable biomaterial differs between rats and mice. AB - Before a biomaterial can be applied in the clinic, biocompatibility must be tested in in vivo models, by monitoring the foreign body reaction. In this study, we compared the foreign body reaction (FBR) to the biodegradable biomaterial hexamethylenediisocyanate crosslinked dermal sheep collagen (HDSC) between several strains of rats and mice. HDSC disks were implanted subcutaneously on the backs of AO, BN, F344, LEW, and PVG rats and on the backs of 129 SVEV, BALB/c, and C57BL/6 mice. Materials were explanted after 7, 14, 21, and 28 days and processed for (immuno) light and transmission electron microscopic evaluation. In all rat strains, giant cell formation and phagocytosis of HDSC bundles were comparable. In addition, in the PVG rat, many plasma cells infiltrated the HDSC disks. Only a few T cells were present in AO and PVG rats, whereas, in F344 and LEW rats, the presence of T cells was more pronounced. BN rats showed an intermediate T-cell infiltration. In mice, the FBR to HDSC was comparable between the different strains. Compared with rats, giant cell formation was limited, whereas stroma formation was more abundant. Phagocytosis of HDSC bundles rarely occurred in mice, whereas calcification was observed more often. It is concluded that the FBR to HDSC clearly differs between rats and mice. This has consequences for assessment studies on biocompatibility and also on fundamental biomaterial research. PMID- 11007612 TI - Surface morphology and wear mechanisms of four clinically relevant biomaterials after hip simulator testing. AB - The surfaces of worn components hold clues to the underlying wear mechanisms. Previous evidence suggested that the absolute wear rates of acetabular components in a hip simulator were related to mechanical behavior; we hypothesized that the surface morphology of the liners might also be sensitive to mechanical properties. A noncontact, three-dimensional surface topography measurement system based on white light interferometry was used to quantify the surface morphology of ultra-high molecular weight polyethylene, polytetrafluoroethylene, high density polyethylene, and polyacetal liners, and their corresponding femoral heads, after 3 million cycles in a multi-directional hip simulator. Comparisons were made with the fatigue soaked and control (as machined) components. Statistically significant power law relationships were observed between the arithmetic mean surface roughness (R(a)) of the worn acetabular liners and the volumetric wear rate in the hip simulator (p < 0.01, r(2) = 0.52). Significant relationships were also observed between R(a) and the elastic and large deformation mechanical behavior of the liner materials, measured directly from the wear-tested liners using the small punch test (p < 0.01, r(2) = 0.54-0.81). The results support the hypothesis that wear mechanisms of acetabular liners during hip simulator testing are related to surface morphology in conjunction with the mechanical behavior of the polymeric materials. PMID- 11007613 TI - The expression of bone matrix protein mRNAs around beta-TCP particles implanted into bone. AB - Tissue response around beta-tricalcium phosphate (beta-TCP) particles (150-300 microm in diameter) implanted into rat tibiae was analyzed by in situ hybridization with digoxigenin-labeled procollagen alpha1(I) (COL), osteonectin, osteocalcin, and osteopontin (OPN) RNA probes. Specimens were collected at 3, 5, 7, and 10 days after the operation. Holes without implantation were used as control. In both the beta-TCP implanted and control groups, new bone was formed centripetally and all four kinds of mRNA were expressed in activated osteoblasts. A COL signal was expressed most strongly and widely, and was detected at the peripheral region of the hole at day 3. The other three mRNAs were also expressed in bone forming osteoblasts by day 7. However, in the earlier cell reaction stage, OPN expression in the beta-TCP implanted group was different than that in the control group: OPN mRNA was seen exclusively in the cells on the particles, and an OPN signal was detected not only in COL-positive cells, but also in COL negative cells. The former cells may be osteoblasts and reflect the early process of bone formation on biomaterials. The latter cells may be macrophages and reflect foreign body reactions. Expression of these OPN mRNAs induced by implantation of beta-TCP may play a role in bone formation on the materials and in determining their biocompatibility. PMID- 11007614 TI - Biocompatibility of electroactive polymers in tissues. AB - The biocompatibility of ethylene-vinyl acetate copolymer (EVAc), polyethylene (PE), and polyaniline (PANi) films in the emeraldine (EM), nigraniline (NA) and leucoemeraldine (LM) intrinsic oxidation states were assessed through subcutaneous implantation into male Sprague-Dawley rats beneath the dorsal skin, for a period ranging from 19 to 90 weeks. Histological examination, interstitial pressure measurement, and X-ray photoelectron spectroscopy (XPS) were employed to determine the biocompatibility of the polymers. The polymers did not provoke inflammatory responses in the subcutaneous tissues over the entire implantation period. Characteristics features associated with tissue-implant incompatibility were not evident near the implantation. Interstitial pressure was measured to evaluate the development of tissue. Low interstitial pressure readings on the region of implantation confirmed the biocompatibility of these polymer types. The surface composition of the electroactive aniline polymers before and after the implantation was characterized by XPS. PMID- 11007615 TI - Biocompatibility evaluation of dental metal alloys in vitro: expression of extracellular matrix molecules and its relationship to cell proliferation rates. AB - The biocompatibility in vitro of dental biomaterials has been widely studied, with consideration of cell viability and cell proliferation rates. In the present study we evaluated the biocompatibility in vitro of three single-phase dental metal alloys, all provided by the same manufacturer. To this aim, we considered the percentage of proliferating cells revealed by 5-bromodeoxyuridine incorporation in human fibroblast cultures in the presence of these biomaterials, performing a short time test (72 h). These data were correlated with immunocytochemical expression of four molecules of the extracellular matrix, i.e., fibronectin, type I collagen, beta(1)-integrin subunit, and chondroitin sulfate, because the capability of cells to adhere to substrata is widely related to cell proliferation rates. Alloys presenting higher amounts of noble elements were more biocompatible even when they contained significant amount of both Ag and Cu. As regards the expression of the extracellular matrix molecules, the organization level of fibronectin in fibrils was correlated with higher cell proliferation rates, whereas no difference was detected for the expression of the other antigens. On these bases, we assume that expression of fibronectin could be a useful parameter in evaluation of biocompatibility in addition to cell proliferation capability. PMID- 11007616 TI - Cyclic fatigue-crack propagation in sapphire in air and simulated physiological environments. AB - Single-crystal aluminas are being considered for use in the manufacture of prosthetic heart valves. To characterize such materials for biomedical application, subcritical crack growth by stress corrosion (static fatigue) and by cyclic fatigue has been examined in sapphire along (1100) planes in 24 degrees C humid air and 37 degrees C Ringer's solution (the latter as a simulated physiological environment). The relationships between crack-propagation rates and the linear-elastic stress intensity have been determined for the first time in sapphire for both modes of subcritical cracking. It was found that growth rates were significantly faster at a given stress intensity in the Ringer's solution compared to the humid air environment. Mechanistically, a true cyclic fatigue effect was not found in sapphire as experimentally measured cyclic fatigue-crack growth rates could be closely predicted simply by integrating the static fatigue crack growth data over the cyclic loading cycle. PMID- 11007617 TI - Novel beta-emitting poly(ethylene terephthalate) surface modification. AB - Restenosis after percutaneous interventions in coronary and peripheral arteries leads to repeat procedures and surgery in a significant number of patients. We have previously demonstrated that irradiation of an arterial site using an endovascular source (brachytherapy) is highly effective in preventing the restenotic process. To this end, a novel beta radiation delivery system was developed, based on the adsorption of (32)P (o-phosphoric acid) by pH-sensitive chitosan hydrogel on a poly(ethylene terephthalate) (PET) balloon surface. The PET balloon surface was treated with oxygen plasma and coated with chitosan hydrogel. Covalent bonds, ionic bonds, and hydrogen bonds all contribute to the adhesion between chitosan hydrogel and PET. In the aqueous phosphoric acid (PA) solution, the -NH(2) groups of chitosan were protonated by PA and the adsorption of PA occurred at the same time. The effect of PA concentration and temperature on adsorption efficiency and kinetics were studied. More than 70% PA was adsorbed on the sample surface in 0.2 mM PA solution. The surface of samples was also investigated by attenuated total reflection-Fourier transform infrared spectroscopy and scanning electron microscopy. PET surface may be modified to carry high activity beta emitters; such materials may be useful in a therapeutic setting PMID- 11007618 TI - Evaluation of calcium phosphates and experimental calcium phosphate bone cements using osteogenic cultures. AB - In this study, rat bone marrow cells (RBM) were used to evaluate two biodegradable calcium phosphate bone cements and bioactive calcium phosphate ceramics. The substances investigated were: two novel calcium phosphate cements, Biocement F and Biocement H, tricalcium phosphate (TCP), surface-modified alpha tricalcium phosphate [TCP (s)] and a rapid resorbable calcium phosphate ceramic consisting of CaKPO(4) (sample code R5). RBM cells were cultured on disc-shaped test substrates for 14 days. The culture medium was changed daily and also examined for calcium, phosphate, and potassium concentrations. Specimens were evaluated using light microscopy, and morphometry of the cell-covered substrate surface, scanning electron microscopy, and energy dispersive X-ray analysis and morphometry of the cell-covered substrate surface. Areas of mineralization were identified by tetracyline labeling. Except for R 5, rat bone-marrow cells attached and grew on all substrate surfaces. Of the different calcium phosphate materials tested, TCP and TCP (s) facilitated osteoblast growth and extracellular matrix elaboration to the highest degree, followed by Biocements H and F. The inhibition of cell growth encountered with R 5 seems to be related to its high phosphate and potassium ion release. PMID- 11007619 TI - Effect of anti-inflammatory medication on monocyte response to titanium particles. AB - Cytokines produced by macrophages in the periprosthetic membranes surrounding joint replacements have been implicated as causal agents in osteolysis and prosthetic loosening. The present study characterizes the response of human peripheral blood monocytes to titanium particles. Monocytes were obtained from donated blood and were cultured in the presence of different-sized titanium particles. Exposure to titanium-aluminum-vanadium particles significantly changed the release of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and interleukin-1 (IL-1), whereas there was no significant effect on the release of prostaglandin E(2) (PGE(2)). When monocytes were cultured with particles, the titanium alloy particles induced significantly more release of TNF-alpha and less IL-1 secretion. Ciprofloxacin inhibited production of TNF-alpha, IL-6, IL-1, and PGE(2) in human monocytes exposed to titanium particles. In contrast to ciprofloxacin, indomethacin was not a potent inhibitor of TNF-alpha production but potentiated IL-6 production in titanium-stimulated monocytes. Indomethacin had no effect on the production of IL-1 and was a potent inhibitor of PGE(2) production in titanium-stimulated monocytes. Pentoxifylline had an inhibitor effect on TNF-alpha production in titanium-stimulated monocytes. Pentoxifylline potentiated IL-6 and IL-1 production in monocytes exposed to titanium particles and had a biphasic effect on the PGE(2) production. The results of this study support our hypothesis that human monocytes release bone resorption mediators after in vitro exposure to TiAlV alloy particles. The results also demonstrate the differences of bone-resorbing mediators in response to different wear particle size. The pharmacologic agents (ciprofloxacin, pentoxifylline, and indomethacin) that can modulate the release of bone resorbing mediators such as PGE(2), TNF-alpha, IL-1, and IL-6 release from human monocytes. The results help to elucidate the differences in cellular response to wear particles but may not be directly transposed to the human situation. PMID- 11007620 TI - Acute cellular interaction with textured surfaces in blood contact. AB - Textured blood-contacting surfaces can promote the formation of a blood compatible pseudo-neointima. We hypothesized that by controlling the surface texturing, the pseudo-neointima thickness could be controlled. The hypothesis was tested experimentally by fabricating the polyurethane textured surfaces with three different fiber lengths, and exposing them simultaneously to the flowing blood in an ovine ex vivo carotid-jugular series shunt for periods up to 4 h. The textured surface consisted of regularly spaced tapered micro-fibers of defined length on a smooth base-plane surface. Because of the simple surface topography, detailed computational fluid-dynamic modeling of the surface could be obtained as a parallel study. Experimental results showed that white cell was the predominant cell type deposited on the textured surfaces, whereas macroscopic thrombus formation occurred only in one of nine blood-contacting experiments. White cell density on the textured base-plane surface was subsequently quantified by image analyzing the electron micrographs of blood-contacted textured surfaces. The statistical analysis of cell densities on individual textured surfaces showed effects of wall shear stress on the textured base plane (which was obtained from the fluid-dynamic modeling), the longitudinal position of the test section in the series shunt, and blood-contact time. PMID- 11007621 TI - Tissue reactions after subcutaneous and intraosseous implantation of mineral trioxide aggregate and ethoxybenzoic acid cement. AB - Biocompatibility of mineral trioxide aggregate and ethoxybenzoic acid cement was investigated by subcutaneous and intraosseous implantation of the materials in rats. Tissue reactions were studied at 15, 30, and 60 days after implantation. Subcutaneous implantation of mineral trioxide aggregate initially elicited severe reactions with coagulation necrosis and dystrophic calcification; the reactions, however, subsided to mostly moderate with time. Subcutaneous implantation of ethoxybenzoic acid cement initially elicited mostly moderate reactions that subsided to mild in time. Osteogenesis was not observed with either material upon subcutaneous implantation indicating that neither material is osteoinductive. Reactions to intraosseous implants of both materials were less intense than with subcutaneous implantation. Osteogenesis occurred in association with intraosseous implants indicating that both materials are osteoconductive. PMID- 11007622 TI - Modulation of mechanical properties in multiple-component tissue adhesives. AB - In vitro, ex vivo, and in vivo studies were performed to investigate the effect of mixing upon the mechanical properties of a two-component tissue adhesive. The hypothesis investigated was that a more complete mixing of the two components would yield an increase in the mechanical performance of the adhesive. This in turn would be demonstrated by improved outcomes in models of clinical sealant application. In vitro stereological analysis of tissue adhesive mixed and delivered by several different applicators demonstrated variation in the amount of mixing provided by each type of delivery system. Ex vivo tensile adhesive strength showed that there was a correlation between the amount of mixing and bonding strength; that is, more thorough mixing demonstrated higher adhesive strength. No significant difference was seen, however, between the different applicator types and impact on in vivo dermal incisional closure strength. There was a correlation, though, in amount of mixing and in vivo hemostasis. In a rabbit spleen incision model, a more thoroughly mixed sealant corresponded with a decrease in time to obtain complete hemostasis, as well as less sealant used. The effects of mixing on tissue-adhesive mechanical performance were influenced somewhat by the amount of mixing provided by the applicator. This effect, however, was dependent upon the sealant formulation and the type of in vivo application. PMID- 11007623 TI - The influence of transforming growth factor beta1 on mesenchymal cell repair of full-thickness cartilage defects. AB - To repair full-thickness articular cartilage defects in rabbit knees, we transplanted periosteal cells in a fibrin gel and determined the influence of transforming growth factor beta (TGF-beta) in vitro. Alginate served as a temporary supportive matrix component and was removed prior to transplantation. The defects were analyzed macroscopically, histologically, and electron microscopically, and evaluated with a semi-quantitative score system. Periosteal cell transplants showed a chondrogenic differentiation, which results in the development of embryonic-like cartilage tissue after 4 weeks and complete resurfacing of the patellar groove after 12 weeks. In the control groups, no repair was observed. Under the influence of TGF-beta1 we observed a reduction of the cartilage layer, whereas the osteochondral integration and the zonal architecture were improved. Periosteal cell-beads are stable cartilage transplants and have stiffness and elasticity enough for easy and sufficient transplant fixation. Further investigations are necessary to optimize the application of TGF-beta1 for cartilage repair. PMID- 11007624 TI - Bioactive macroporous titanium surface layer on titanium substrate. AB - A macroporous titanium surface layer is often formed on titanium and titanium alloy implants for morphological fixation of the implants to bone via bony ingrowth into the porous structure. The surface of titanium metal was recently shown to become highly bioactive by being subjected to 5.0 M-NaOH treatment at 60 degrees C for 24 h and subsequent heat treatment at 600 degrees C for 1 h. In the present study, the NaOH and heat treatments were applied to a macroporous titanium surface layer formed on titanium substrate by a plasma spraying method. The NaOH and heat treatments produced an uniform amorphous sodium titanate layer on the surface of the porous titanium. The sodium titanate induced a bonelike apatite formation in simulated body fluid at an early soaking period, whereby the apatite layer grew uniformly along the surface and cross-sectional macrotextures of the porous titanium. This indicates that the NaOH and heat treatments lead to a bioactive macroporous titanium surface layer on titanium substrate. Such a bioactive macroporous layer on an implant is expected not only to enhance bony ingrowth into the porous structure, but also to provide a chemical integration with bone via apatite formation on its surface in the body. PMID- 11007625 TI - Possible explanation for the white band artifact seen in clinically retrieved polyethylene tibial components. AB - Studies have focused attention on the appearance of a subsurface white band in clinically retrieved polyethylene components and the possible contribution of this phenomenon to early polyethylene delamination. Unconsolidated polyethylene particles and oxidation have been suggested as possible reasons for the appearance of the white band. Calcium stearate and other additives used in processing ultra-high molecular weight polyethylene may also contribute to formation of the white band. A quantitative investigation was conducted on 11 retrieved tibial components that exhibited a subsurface white band to determine whether the amount of calcium stearate particles and additives were greater in the white band region when compared with the mid-portion of the same section of polyethylene. Calcium stearate particles and other additives were quantified using backscattered electron imaging with correlated elemental analysis. The particles were identified based on morphology and elemental patterns similar to reference calcium stearate particles and known additives. Significantly more (p < 0. 0001) calcium stearate particles and additives were present in the white band region (4578 +/- 418 particles/mm(2); mean +/- standard error) than the mid portion region (1250 +/- 147 particles/mm(2)) of the sectioned tibial inserts. The percent area occupied by calcium stearate particles and additives was five times higher (p < 0.0001) within the white band region (0.81 +/- 0.10%) than the mid-portion region (0.16 +/- 0.03%). The increased presence of calcium stearate and other additives in the white band region suggests that they may play a role in the formation of the white band. In future investigations it may be important to consider how calcium stearate and other additives in polyethylene resins affect white band formation and the possible contribution to crazing, early delamination, and osteolysis in total joint replacement. PMID- 11007626 TI - Adsorption of peroxidase on titanium surfaces: A pilot study. AB - The present study demonstrates the in vitro and in vivo adsorption of peroxidase onto titanium surfaces. Titanium foils (mean +/- SEM: 365 +/- 2 mm(2), n = 114) were incubated during 30 min with lactoperoxidase (4 mg in 5 mL 100 mM phosphate buffer pH 7). After 15 washings by H(2)O, titanium foils were incubated with o phenylenediamine (6 mg/mL) and H(2)O(2) (7 mM) during 30 min. The reaction was then stopped by the addition of HCI 1M and the absorbance of the liquid phase was read on a spectrophotometer at 492 nm. In vitro adsorbed lactoperoxidase onto titanium surfaces was 0.70 +/- 0.05 ng/mm(2) (mean +/- SEM, n = 30). X-ray photoelectron spectroscopy confirmed the incorporation of protein nitrogen onto titanium surfaces: the nitrogen atomic percentage increased from 0.9 +/- 0.3 to 12.7 +/- 0.2% (n = 3) and from 3.7 +/- 0.1 to 14.4 +/- 0. 4% (n = 5) when titanium foils were incubated in the lactoperoxidase solution during 30 min and 24 h respectively. In vivo, oral peroxidases adsorbed on titanium healing abutments from 0.01 to 0.58 ng/mm(2) (n = 19) after 2 weeks in the oral environment. PMID- 11007627 TI - Effect of vapor-flame treatment on plasma sprayed hydroxyapatite coatings. AB - A vapor-flame treatment was developed to modify the crystallinity of the as sprayed hydroxyapatite (HA) coatings. The effects of the treatment on composition, structure, and properties of HA coatings were investigated. Results showed that the vapor-flame treatment is simple and efficient to adjust the crystallinity of as-sprayed HA coating. Its crystallinities can be raised from 53.5 to 98.7% in 3-7 min. The porosities of coatings increased with an increase in the vapor-flame treating time. The microhardness of coating decreased as a result of this treatment. It may be explained in terms of the extent of microcracks caused by recrystallization of amorphous HA and relaxation of stress of the coating. The porosity, bonding strength, and hardness of HA coatings treated for 7 min were 15.7%, 32.0 MPa (300 microm thickness), and 1.9 GPa, respectively. PMID- 11007628 TI - Comparative study of seeding methods for three-dimensional polymeric scaffolds PMID- 11007629 TI - Total intravenous anesthesia with midazolam, remifentanil, propofol and cistracurium in morbid obesity. AB - BACKGROUND: According to physical impairments of massive obesity, cardiac, respiratory and gastrointestinal physiology must be considered as much as pharmacokinetic behavior. Anesthetic management of morbidly obese patients has to be carefully planned, in order to minimize the increased risks of aspirative pneumonitis, hemodynamic instability and delay in recovery. The ideal anesthesia should provide a smooth and quick induction, allowing rapid airway control, prominent hemodynamic stability, and rapid emergence from anesthesia. To approach these ideal conditions, a Total Intravenous Anesthesia (TIVA) with midazolam, remifentanil, propofol and cisatracurium was designed and analyzed. METHODS: 10 consenting morbidly obese patients scheduled for elective Laparoscopic Adjustable Gastric Banding participated in the study. TIVA with midazolam, remifentanil, propofol and cisatracurium was used in all cases. Time to loss of consciousness, tracheal intubation, perianesthetic physiological parameters and complications, incidence of awareness with recall, recovery times, postoperative analgesia and costs of drugs were evaluated. RESULTS: The analyzed data showed adequate time and physiological conditions for induction and tracheal intubation, stable maintenance with easy handling of deepness, low incidence of perianesthetic complications, excellent recovery performance and institutional efficiency. CONCLUSIONS: TIVA with midazolam, remifentanil, propofol and cisatracurium was found to be effective, secure, predictable and economic for the anesthetic management of morbidly obese patients. PMID- 11007630 TI - Laparoscopic Roux-en-Y gastric bypass: transoral or transgastric anvil placement? AB - BACKGROUND: Laparoscopic techniques have been used to perform the Roux-en-Y gastric bypass (RYGBP). The gastrojejunostomy may be constructed using an end-to end anastomosis (EEA) stapler. Most reports describe passing the EEA anvil transorally using an esophagogastroscope and a pull-wire technique. METHOD: We describe problems experienced using this technique and present an alternative method. RESULTS: Esophageal injury may occur during laparoscopic RYGBP (LRYGBP) using the transoral anvil placement technique. When the anvil is retrieved into the gastric pouch, the anvil may become lodged at the cricopharngeus muscle. Dislodgment can be problematic and time-consuming. We present a case of mild esophageal injury which occurred during transoral anvil placement. The patient had transient postoperative dysphagia and recovered without sequelae. We present an alternative method in which the anvil is passed through a gastrotomy. CONCLUSION: Transgastric anvil placement alleviates the need for endoscopy, thereby saving time and resources. This technique eliminates the potential for esophageal injury. The transgastric anvil placement technique has proven reliable. The transgastric method may make the LRYGBP operation safer and easier to perform. PMID- 11007631 TI - Silastic ring vertical gastroplasty by minilaparotomy. AB - BACKGROUND: The incision length has significant influence on the postoperative course after silastic ring vertical gastroplasty (SRVG). METHODS: A technique is described in which SRVG can be performed through a minilaparotomy incision. RESULTS: 110 patients were operated with this technique during 1996. No remarkable intraoperative or postoperative complications were encountered. Postoperative weight loss has been satisfactory. CONCLUSIONS: Performance of SRVG through a minilaparotomy is feasible. The technique is recommended for surgeons familiar with the operation through the formal incision. PMID- 11007632 TI - Laparoscopic silicone adjustable gastric band: initial experience in finland. AB - BACKGROUND: The Swedish adjustable gastric band (SAGB) was introduced in 1985 and rapidly gained popularity. Today more than 21,000 gastric banding procedures have been performed in Europe. The reported results of gastric banding operations are mainly good, although the method is not without controversies and risks. We report here our initial experience with the SAGB. METHODS: 60 patients (44 women, 16 men) were treated surgically for morbid obesity between the years 1996 and 1999, with SAGB. Median age of the patients was 44 years (range 21-64) and preoperative median Body Mass Index (BMI, kg/m(2)) was 45 (range 35-55). 3 patients were operated by an open approach, and the remaining 57 laparoscopically. RESULTS: Operative time was 62-206 minutes (median 97 minutes). Only one operation was converted to open approach (1.8%), due to extensive adhesions. No intraoperative complications occurred. At 1 year follow-up, mean weight loss was 30 kg, mean excess weight loss was 50%, and median BMI was 35. 4 patients have been reoperated so far (6.7%) due to slippage of the band (2 patients), infection of the band (1 patient), and leaking of the filling system (1 patient). Median postoperative hospital stay was 3 days (range 2-53). Mortality was 0%. Immediate postoperative morbidity-rate was 12% (7/60), although serious morbidity occurred in only 1 patient (1.7%). CONCLUSIONS: Laparoscopically placed adjustable gastric band is a good option for the morbidly obese patient. PMID- 11007633 TI - Influence of metabolic state and diabetes on the outcome at the end of first year after gastric banding. AB - BACKGROUND: The influence of metabolic state and the presence of diabetes before surgery on the weight changes following non-adjustable gastric banding, were studied. METHODS: The total of 50 patients referred for gastric banding had the following parameters measured: insulinemia, glycemia, total cholesterol, triglycerides (TAG), dehydroepiandrostendione (DHEA) and its sulphate (DHEA-S), triiodothyronine, thyroxine, and thyroxine-stimulating hormone. We evaluated 28 of these, who completed at least 6 months of follow-up. 12 of these patients had diet-treated non-insulin dependent diabetes mellitus (NIDDM) and 16 were non diabetics, and 9 of them had a positive family history of NIDDM (FH DM). We compared the change in weight of the diabetics, non-diabetics and those with positive FH DM, at 3, 6 and 12 months after the operation. Multiple linear regression tested the influence of the measured parameters on patients' weight. RESULTS: Weight loss was greatest in the group of patients without a positive family history of diabetes, and lowest in the diabetic group. Due to the high dispersion, differences in the t-test are not significant. When individual factors and their influence on weight development were tested by multiple linear regression, weight loss was greatest in patients with high TAG and low insulin levels and lowest in patients with diabetes or positive FH DM. CONCLUSIONS: Although post-operative weight can be influenced by other factors, eg. psychological ones, it is advisable to test each patient pre-operatively for insulin and TAG levels, and to establish family history of diabetes and presence of diabetes to give an idea of the prognosis of weight change. PMID- 11007634 TI - Laparoscopic gastric bypass: endostapler transoral or transabdominal anvil placement. PMID- 11007635 TI - Warning signals, receiver psychology and predator memory. AB - This review identifies four receiver psychology perspectives that are likely to be important in the design and evolution of warning signals. Three of these perspectives (phobia, learning and prey recognition) have been studied in detail, and I include a brief review of recent work. The fourth, a memory perspective, has received little attention and is developed here. A memory perspective asks, 'how might warning signals function to reduce forgetting of avoidances between encounters?'. To answer this question I review data from psychology literature that describe important features of animal long-term memory. These data suggest that components of warning signals may function to reduce forgetting (and therefore increase memorability) by (1) preventing forgetting of learnt prey discriminations; (2) jogging the memories of forgetful predators; and (3) biasing forgetting in favour of prey avoidance when the warning signal of a defended aposematic species is copied by an edible Batesian mimic. A combination of a learning and a memory perspective suggests that the features of aposematic prey that accelerate avoidance learning may also be the features that decelerate forgetting processes. If correct, this would have important implications for the comprehension of signal design. Finally, I suggest that the cryptic appearance of an edible prey may decelerate predator learning and accelerate predator forgetting, to the benefit of the prey. In terms of learning and memory, crypsis may be an antisignal. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007636 TI - Re-expression of songs deleted during vocal development in white-crowned sparrows, Zonotrichia leucophrys. AB - White-crowned sparrows learn and produce multiple song types as juveniles, but most individuals stop singing all except one by the end of the first singing season. This single song type is generally maintained throughout adulthood. We demonstrate that, at the start of the second and subsequent singing seasons, this species can recall songs that had been deleted during the first singing season. The re-expression of song occurred in both the oriantha and the gambelii subspecies. Although all our males recrystallized the original song in the second year, our results indicate a mechanism for seasonal song change without new song memorization. The traditional dichotomy of closed-ended versus open-ended learning is inadequate for birds that learn early in life but can change their song output seasonally. We suggest that species can exhibit a closed sensitive period for song memorization and first production, with the ability to recall deleted songs later in life. This type of learning, selective attrition followed by subsequent re-expression, may be used by some species currently considered open-ended learners. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007637 TI - MHC-mediated fetal odourtypes expressed by pregnant females influence male associative behaviour. AB - Mice can recognize one another by individually characteristic phenotypic body odours (odourtypes) that reflect their genetic constitution at the highly polymorphic major histocompatibility complex (MHC) of genes on chromosome 17. We have shown previously that MHC-determined odours are produced by fetuses: house mice, Mus domesticus, can be trained to discriminate between genetically identical pregnant females carrying 9-18-day-old fetuses of differing MHC type. Theoretically, it should be possible for a mouse to determine the MHC type of the sire based on the odourtype of the pregnant female. In the current study we investigated whether untrained male mice show spontaneous discrimination between such pregnant mice. In experiment 1, sexually inexperienced male mice spent more time near pregnant females that carried fetuses most genetically different from the males themselves. Experiment 2, designed to evaluate possible experiential effects on this preference, tested males that were cohabiting and had impregnated a female that was either genetically identical to the test male (excepting X and Y chromosomes) or differed from him only at the MHC. Males in the former case performed virtually identically to those tested in experiment 1. In contrast males in the latter group did not display this preference. These studies reveal that among untrained male mice, fetal MHC type influences choice behaviour presumably via fetal odourtypes expressed in maternal secretions/excretions and that previous housing and/or mating experience modulates male choice. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007638 TI - A species-specific acoustic cue for selective song learning in the white-crowned sparrow. AB - Song learning in birds is paradoxical. Without tutoring, songbirds do not develop normal songs. Yet despite this inability, birds possess extensive foreknowledge, in a mechanistic sense, about the normal song of their species. When given a choice of tape recordings, young, naive songbirds select sounds of their own species for imitation. We tape-tutored white-crowned sparrows, Zonotrichia leucophrys oriantha, with a set of manipulated songs to investigate whether the introductory whistle universally present in white-crowned sparrow song guides selective song learning in this species. Our results confirm that this whistle serves as a cue for song learning, enabling acquisition of normally rejected sounds of other species, including hermit thrush, Catharus guttatus, notes, which have a sound quality distinct from that of natural white-crowned sparrow phrases. Our results support the conclusion that sensory mechanisms rather than motor constraints are primarily responsible for the selectivity seen in song learning. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007639 TI - Survival regression analysis: a powerful tool for evaluating fighting and assessment. AB - Theoretical models of animal contests frequently generate predictions about how asymmetries (e.g. differences in size, residence status) between contestants affect fight duration. Linear regression and nonparametric correlation analyses are commonly used to test the fit of data to such models. We show how survival regression analysis (SRA) is a powerful technique for studying the effect of asymmetries on the duration of contests. SRA, which is under-utilized by students of animal behaviour, offers several advantages over more frequently used procedures. It provides unbiased parameter estimates even when including censored data (i.e. results of contests that have not ended at the time when observations are stopped). The analysis of hazard functions, which is a component of SRA, is an easy way to test for consistency with predictions of the sequential assessment game model. These and other advantages of SRA are illustrated by using SRA and more conventional methods to analyse the effect of asymmetries on contest duration for encounters between female Mediterranean tarantulas, Lycosa tarentula (L.). It is hoped that this example of the advantages of SRA will encourage more widespread use of this powerful technique. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007640 TI - Chemically mediated predator inspection behaviour in the absence of predator visual cues by a characin fish. AB - Animals commonly approach (i.e. 'inspect') potential predators. Glowlight tetras, Hemigrammus erythrozonus, have previously been shown to inspect the combined chemical and visual cues originating from novel predators and to modify their inspection (approach) behaviour depending upon the predator's diet. We conducted two experiments to determine whether tetras would inspect the chemical cues of injured prey or the dietary cues of a novel predator in the absence of any visual cues. Shoals of glowlight tetras were exposed to either distilled water (control) or the skin extract of swordtail (lacking ostariophysan alarm pheromones) or the skin extract of tetra (with alarm pheromones). There was no significant difference in the frequency of predator inspection behaviour towards swordtail or tetra skin extract compared to the distilled water controls. In the second experiment, we exposed shoals of tetras to either distilled water or the odour of Jack Dempsey cichlids, Cichlasoma octofasciatum, which had been food deprived, or fed a diet of swordtails or tetras. There was no significant difference in the frequency of predator inspection behaviour towards the odour of the starved cichlids and the odour of the fed cichlids in either of the two diet treatments. However, when tetras were exposed to the odour of cichlids fed tetras, they took significantly longer to initiate an inspection visit, remained further from the source of the chemical cues and inspected in smaller groups, compared with the odour of a starved cichlid or a cichlid fed swordtails. These data strongly suggest that tetras will inspect chemical cues alone, but only if the cue contains information about the predator. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007641 TI - Mechanisms of maternal rank 'inheritance' in the spotted hyaena, Crocuta crocuta. AB - Maternal rank 'inheritance', the process by which juveniles attain positions in the dominance hierarchy adjacent to those of their mothers, occurs in both cercopithecine primates and spotted hyaenas. Maternal rank is acquired in primates through defensive maternal interventions, coalitionary support and unprovoked aggression ('harassment') directed by adult females towards offspring of lower-ranking individuals. Genetic heritability of rank-related traits plays a negligible role in primate rank acquisition. Because the social lives of Crocuta and cercopithecine primates share many common features, we examined whether the same mechanisms might operate in both taxa to promote maternal rank 'inheritance'. We observed a large clan of free-living spotted hyaenas in Kenya to test predictions of four mechanistic hypotheses. Hyaena rank acquisition did not appear to be directly affected by genetic heritability. Unprovoked aggression from adult female hyaenas was not directed preferentially towards low-ranking cubs. However, high-ranking mothers intervened on behalf of their cubs more frequently and more effectively than low-ranking mothers. Maternal interventions and supportive coalitions appeared to reinforce aggression directed at 'appropriate' conspecific targets, whereas coalitionary aggression directed at cubs apparently functioned to extinguish their aggressive behaviour towards 'inappropriate' targets. Young hyaenas and primates thus appear to 'inherit' their mothers' ranks by strikingly similar mechanisms. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007642 TI - The influence of age and size on temporal mate signalling behaviour. AB - I investigated how male size, condition and age influence both time spent calling and how signals are apportioned throughout the night (i.e. temporal calling pattern) in the Texas field cricket, Gryllus integer. I quantified male calling time and temporal calling pattern using an electronic apparatus that continuously monitored male calling/noncalling behaviour throughout the night. Male condition, measured using the residuals from an allometric regression of male mass on overall body size, did not explain any variation in either time spent calling or temporal calling pattern. However, some intrapopulation variation in time spent calling and temporal calling pattern was explained by differences in male size and age. Large males called more often than small males. Young and very old adult males called significantly less often than middle-age males. As males aged they initiated calling earlier in the evening, probably increasing their susceptibility to parasitism by the tachinid parasitoid Ormia ochracea. Overall, age and size differences explained 10-40% of signalling variation in male G. integer. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007643 TI - Experimental evidence that group foragers can converge on predicted producer scrounger equilibria. AB - When foraging together, animals are often observed to feed from food discoveries of others. The producer-scrounger (PS) game predicts how frequently this phenomenon of food parasitism should occur. The game assumes: (1) at any moment all individuals can unambiguously be categorized as either playing producer (searching for undiscovered food resources) or scrounger (searching for exploitation opportunities), and (2) the payoffs received from the scrounger tactic are negatively frequency dependent; a scrounger does better than a producer when the scrounger tactic is rare, but worse when it is common. No study to date has shown that the payoffs of producer and scrounger conform to the game's assumptions or that groups of foragers reach the predicted stable equilibrium frequency (SEF) of scrounger, whereby both tactics obtain the same payoff. The current study of three captive flocks of spice finches, Lonchura punctulata, provides the first test of the PS game using an apparatus in which both assumptions of the PS game are met. The payoffs to the scrounger, measured as feeding rate (seeds/s), were highly negatively frequency dependent on the frequency of scrounger. The feeding rate for scrounger declined linearly while the rate for producer either declined only slightly or not at all with increasing scrounger frequency. When given the opportunity to alternate between tactics, the birds changed their use of each, such that the group converged on the predicted SEF of scrounger after 5-8 days of testing. Individuals in this study, therefore, demonstrated sufficient plasticity in tactic use such that the flock foraged at the SEF of scrounger. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007644 TI - Using video playbacks to study visual communication in a marine fish, Salaria pavo. AB - Video playbacks have been successfully applied to the study of visual communication in several groups of animals. However, this technique is controversial as video monitors are designed with the human visual system in mind. Differences between the visual capabilities of humans and other animals will lead to perceptually different interpretations of video images. We simultaneously presented males and females of the peacock blenny, Salaria pavo, with a live conspecific male and an online video image of the same individual. Video images failed to elicit appropriate responses. Males were aggressive towards the live male but not towards video images of the same male. Similarly, females courted only the live male and spent more time near this stimulus. In contrast, females of the gynogenetic poecilid Poecilia formosa showed an equal preference for a live and video image of a P. mexicana male, suggesting a response to live animals as strong as to video images. We discuss differences between the species that may explain their opposite reaction to video images. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007645 TI - Individual mares bias investment in sons and daughters in relation to their condition. AB - The Trivers-Willard hypothesis (TWH) predicts that a mother will treat a son or daughter differently depending on her ability to invest and the impact of her investment on offspring reproductive success. Although many studies have investigated the hypothesis, few have definitively supported or refuted it because of confounding factors or an inappropriate level of analysis. We studied maternal investment in sons and daughters in feral horses, Equus caballus, which meet the assumptions of the TWH with a minimum of confounding variables. Population level analyses revealed no differences in maternal behaviour towards sons and daughters. When we incorporated mare condition, we found that sons were more costly to mares in good condition, whereas daughters were more costly to mares in poor condition, although no differences in maternal behaviour were found. However, since the TWH makes predictions about individual mothers, we examined investment by mares who reared both a son and a daughter in different years of the study. Mares in good condition invested more in their sons in terms of maternal care patterns, costs to maternal body condition and costs to future reproduction. Conversely, mares in poor condition invested more in daughters. Therefore, with an appropriate level of analysis in a species in which confounding variables are minimal, the predictions of the Trivers-Willard hypothesis are supported. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007646 TI - Diet of a polyphagous arthropod predator affects refuge seeking of its thrips prey. AB - Antipredator behaviour of prey costs time and energy, at the expense of other activities. However, not all predators are equally dangerous to all prey; some may have switched to feeding on another prey species, making them effectively harmless. To minimize costs, prey should therefore invest in antipredator behaviour only when dangerous predators are around. To distinguish these from harmless predators, prey may use cues related to predation on conspecifics, such as odours released by a predator that has recently eaten conspecific prey or alarm pheromones released by attacked prey. We studied refuge use by a herbivorous/omnivorous thrips, Frankliniella occidentalis, in response to odours associated with a generalist predatory bug, Orius laevigatus, fed either with conspecific thrips or with other prey. The refuge used by thrips larvae is the web produced by its competitor, the two-spotted spider mite, Tetranychus urticae, where thrips larvae experience lower predation risk because the predatory bug is hindered by the web. Thrips larvae moved into this refuge when odours associated with predatory bugs that had previously fed on thrips were present, whereas odours from predatory bugs that had fed on other prey had less effect. We discuss the consequences of this antipredator behaviour for population dynamics. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007647 TI - Effect of maternal and paternal line on spatial and temporal marine distribution in Atlantic salmon. AB - We examined the inheritance of the sea migration pattern of Atlantic salmon, Salmo salar, in a crossing and tagging experiment in the Baltic Sea. Individuals from the parental stocks, Neva and Iijoki, and their reciprocal hybrids were released as 2-year-old smolts, into the same estuary of the Bothnian Sea in 1994. Two thousand smolts from each of the four groups were marked with Carlin tags. The recapture rate of the tags was nearly 10%. We used log-linear models to analyse the marine distribution of the salmon groups from the tag recovery data. The pure stocks and their pooled hybrid groups all showed statistically significant differences between each other in spatial and temporal sea distribution. The Iijoki salmon were more frequently (9%) caught outside the Bothnian Sea than were the Neva salmon (2%). The majority of the Iijoki salmon (55%), but fewer Neva salmon (40%), were caught in the second sea year. In spatial distribution, the hybrids seemed to be intermediate between the parental stocks, with no differences between reciprocal female and male lines. In duration of sea migration and age at maturity, however, the hybrids were very similar to their maternal line, the effect of which was thus clearly stronger than that of the paternal line. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007648 TI - Diversity of behaviour during novel object tests is reduced in pigs housed in substrate-impoverished conditions. AB - Modern intensive farming conditions lack the diversity of substrates present in more natural environments and offer young animals fewer opportunities for interaction. Evidence exists that this may affect the organization of interactive patterns of behaviour, but shifts in behavioural diversity have not been measured directly. We investigated the effect of the substrate in the home pen on the diversity of behaviour in young growing pigs, Sus scrofa. Over 5 months, 26 pigs were housed singly in either substrate-impoverished (SI) or substrate-enriched (SE) conditions. Once every month we recorded the behaviour of these pigs in detail both in the home pens and in two novel object tests. In addition, we calculated the diversity of behaviour shown by SI and SE pigs in the home pen and in the novel object tests, using a relative behavioural diversity index. In the two novel object tests, SI pigs were less mobile than SE pigs and focused their behaviour on particular substrates. In addition, SI pigs showed less diverse behaviour than SE pigs. Our results show that the less diverse behaviour of SI pigs previously recorded in their home pens persists under novel conditions, supporting the hypothesis that substrate-impoverished housing conditions structurally affect the organization of behaviour in young growing pigs. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007649 TI - Individual food-hoarding decisions in a nonterritorial coal tit population: the role of social context. AB - Among the Paridae, food hoarding is thought to be strongly associated with the exclusive use of territories by winter groups, although it has also been described in populations with loose social systems. However, detailed data on such populations are scarce. To identify the mechanisms underlying individual storing decisions, I studied hoarding behaviour in a nonterritorial, high-density coal tit, Parus ater, population in a subalpine forest. The presence of close neighbours (within 5 m) had the strongest, negative influence on caching probability, whereas more distant neighbours foraging in the same flock did not affect the probability of caching. Adults concentrated their stores in the centre of their home ranges. Caching location, but not caching intensity, was affected by the time of day. Coal tits took longer, and travelled further, to cache food in the inner parts of trees than in the outer parts. Resident adults stored in inner, safer caching locations more often than juvenile residents and transients. Differences in hoarding effort according to the storing substrate, and the daily storing location pattern of juvenile residents and transients, suggest that stores have a different use depending on where they are located. Therefore, my results show that hoarding is compatible with a nonterritorial, nonbreeding social system; hoarding could thus have originated in an ancient, nonterritorial, but sedentary, tit species. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007650 TI - Pigs shift too: foraging strategies and spatial memory in the domestic pig. AB - In a previous experiment, we showed that domestic pigs, Sus scrofa, unlike many other species, performed accurately in a spatial memory task, where visits to a previously baited food trough were rewarded (win-stay). We investigated whether pigs have a predisposition for this strategy, by comparing their performance in a radial arm maze under either win-stay (N=10) or win-shift (N=10) reward contingencies. Contrary to our earlier results, only one of the animals in the win-stay condition was able to reach the imposed criterion level of accuracy. The performances of the other win-stay pigs did not deviate from random. All pigs in the win-shift condition reached criterion by day 25 of the experiment, and performed better than expected by chance. Analysis of the types of errors made matched our a priori predictions that shift movements would occur more frequently, especially within visits to the maze. We suggest that the difference in learning rates may reflect the fact that win-stay pigs needed to use two different rules, stay between trials and shift within trials, while win-shift pigs only needed to use the shift rule. In our previous study, win-stay pigs did not experience a conflict of rules and this may have facilitated stay learning. We found evidence of a recency effect in win-shift animals and a primacy effect in the win-stay group. However, we discuss the unsuitability of these specific terms in this type of experiment, and propose an alternative interpretation of the results. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11007651 TI - Crystal structure of Mycobacterium tuberculosis 7,8-dihydropteroate synthase in complex with pterin monophosphate: new insight into the enzymatic mechanism and sulfa-drug action. AB - The enzyme 7,8-dihydropteroate synthase (DHPS) catalyzes the condensation of para aminobenzoic acid (pABA) with 6-hydroxymethyl-7, 8-dihydropterin-pyrophosphate to form 7,8-dihydropteroate and pyrophosphate. DHPS is essential for the de novo synthesis of folate in prokaryotes, lower eukaryotes, and in plants, but is absent in mammals. Inhibition of this enzyme's activity by sulfonamide and sulfone drugs depletes the folate pool, resulting in growth inhibition and cell death. Here, we report the 1.7 A resolution crystal structure of the binary complex of 6-hydroxymethylpterin monophosphate (PtP) with DHPS from Mycobacterium tuberculosis (Mtb), a pathogen responsible for the death of millions of human beings each year. Comparison to other DHPS structures reveals that the M. tuberculosis DHPS structure is in a unique conformation in which loop 1 closes over the active site. The Mtb DHPS structure hints at a mechanism in which both loops 1 and 2 play important roles in catalysis by shielding the active site from bulk solvent and allowing pyrophosphoryl transfer to occur. A binding mode for pABA, sulfonamides and sulfones is suggested based on: (i) the new conformation of the closed loop 1; (ii) the distribution of dapsone and sulfonamide resistance mutations; (iii) the observed direction of the bond between the 6-methyl carbon atom and the bridging oxygen atom to the alpha-phosphate group in the Mtb DHPS:PtP binary complex; and (iv) the conformation of loop 2 in the Escherichia coli DHPS structure. Finally, the Mtb DHPS structure reveals a highly conserved pterin binding pocket that may be exploited for the design of novel antimycobacterial agents. PMID- 11007652 TI - Pitfalls in homozygosity mapping. AB - There is much interest in use of identity-by-descent (IBD) methods to map genes, both in Mendelian and in complex disorders. Homozygosity mapping provides a rapid means of mapping autosomal recessive genes in consanguineous families by identifying chromosomal regions that show homozygous IBD segments in pooled samples. In this report, we point out some potential pitfalls that arose during the course of homozygosity mapping of the enhanced S-cone syndrome gene, resulting from (1) unexpected allelic heterogeneity, so that the region containing the disease locus was missed as a result of pooling; (2) identification of a homozygous IBD region unrelated to the disease locus; and (3) the potential for inflation of LOD scores as a result of underestimation of the extent of inbreeding, which Broman and Weber suggest may be quite common. PMID- 11007653 TI - Chromosomal duplication involving the forkhead transcription factor gene FOXC1 causes iris hypoplasia and glaucoma. AB - The forkhead transcription factor gene FOXC1 (formerly FKHL7) is responsible for a number of glaucoma phenotypes in families in which the disease maps to 6p25, although mutations have not been found in all families in which the disease maps to this region. In a large pedigree with iris hypoplasia and glaucoma mapping to 6p25 (peak LOD score 6.20 [recombination fraction 0] at D6S967), no FOXC1 mutations were detected by direct sequencing. However, genotyping with microsatellite repeat markers suggested the presence of a chromosomal duplication that segregated with the disease phenotype. The duplication was confirmed in affected individuals by FISH with markers encompassing FOXC1. These results provide evidence of gene duplication causing developmental disease in humans, with increased gene dosage of either FOXC1 or other, as yet unknown genes within the duplicated segment being the probable mechanism responsible for the phenotype. PMID- 11007654 TI - Commentary. monitoring tobacco use in canada: the need for a surveillance strategy. AB - Smoking behaviour has been monitored nationally through population surveys for 35 years in Canada, but these surveys have not been as consistent or rigorous as the magnitude of the smoking problem demands. Inconsistent methods and irregular survey intervals are just two of the characteristics that have made it difficult to know exactly how smoking is changing. Further, an absence of routine data on tobacco control policies (other than the price of cigarettes) has hampered understanding of the determinants of changing prevalence. The advent of two survey series--Canadian Tobacco Use Monitoring Survey (CTUMS) and Canadian Community Health Survey (CCHS) promises to change this situation for the better. We suggest that both are critical elements of a national smoking surveillance system and that, with a commitment to CTUMS in particular, Health Canada could set a new international standard for surveillance. PMID- 11007655 TI - Predictors of smoking cessation in an incentive-based community intervention. AB - The Quit and Win Challenge, an incentive-based intervention, was implemented in two counties in Eastern Ontario to encourage adult smokers to quit smoking. Participants (n = 231) were compared with adult smokers selected at random (n = 385) from a larger, four-county area. Baseline characteristics were assessed by telephone interview, including socio-demographic and smoking-related factors. Follow-up interviews were also conducted by telephone. Initial and follow-up response rates were high (over 84%) in both groups. Compared with the random survey group, Quit and Win participants tended to be younger, more educated, employed and heavier smokers, with fewer friends or co-workers who smoked. After one year, 19.5% of them reported that they were smoke-free, whereas less than 1% of the random group had achieved cessation. This translates into an impact rate of 0.17%, affecting 1 in 588 adult smokers. With the exception of the smokers' baseline "stage of change," none of the socio-demographic or smoking factors was predictive of cessation. We conclude that this intervention achieved only limited success and attracted certain sectors of the community disproportionately, i.e. smokers who were highly motivated to quit. We argue that increased access to proven cessation therapies would improve the impact of such interventions. PMID- 11007656 TI - School-based smoking prevention: economic costs versus benefits. AB - The objective of this study was to conduct a cost-benefit analysis to compare the costs of developing and delivering an effective school-based smoking prevention program with the savings to be expected from reducing the prevalence of smoking in the Canadian population over time. A smoking prevention program that meets published criteria for effectiveness, implemented nationally in Canada, would cost $67 per student (1996 dollars). Assuming such a program would reduce smoking by 6% initially and 4% indefinitely, lifetime savings on health care would be $3,400 per person and on productivity, almost $14,000. The benefit-cost ratio would be 15.4 and the net savings $619 million annually. Sensitivity analyses reveal that considerable economic benefits could accrue from an effective smoking prevention program under a wide range of conditions. PMID- 11007657 TI - Performance of the composite international diagnostic interview short form for major depression in a community sample. AB - Recently, short-form versions of structured psychiatric diagnostic interviews have been developed for epidemiologic and survey research. These short forms can reduce research costs in large-scale studies; however, their accuracy is likely to be less than that of a full diagnostic interview. We evaluated the positive and negative predictive values of a short-form interview derived from the Composite International Diagnostic Interview (CIDI). Samples of subjects who scored both positively (n = 277) and negatively (n = 136) on the CIDI Short Form for Major Depression (CIDI-SFMD) were administered the full depressive disorders section of the CIDI. Almost all subjects who were negative on the short form were similarly classified as not having major depression by the CIDI. Approximately 25% of subjects had false positive results; these subjects tended to be older and less educated than true positives. Approximately 75% of subjects scoring five or more on the CIDI-SFMD had major depression according to the full CIDI, and a proportion of the remainder had less severe depressive syndromes. Some CIDI- SFMD positive subjects may have had depressive symptoms attributable to organic or other etiologies excluded under the definition of major depression. PMID- 11007658 TI - Health-adjusted life expectancy at the local level in ontario. AB - Health expectancy measures are becoming a common method of combining information on mortality and health-related quality of life into one summary population health measure. However, health expectancy measures are infrequently measured at the local level, despite a shift toward health service planning to that level. Using a modified Sullivan method, we calculated health-adjusted life expectancy (HALE) for the 42 public health units in Ontario using life tables that were derived from mortality and population data for 1988-1992 and the Health Utilities Index from the 1990 Ontario Health Survey. There were large variations among health units in HALE at age 15 for both men (range: 51.3-58.2 years) and women (range: 56.6-62.9 years). Generally, rural and northern areas had the lowest HALE. Local differences in male HALE were greater than for life expectancy (7.1 versus 6.0 years). Despite a relatively large health survey (45,583 respondents, range: 729-1,746 per health unit), few HALE differences deviated significantly from the Ontario mean, raising concerns about the feasibility of estimating local health expectancy measures with adequate precision. Nevertheless, the wider local differences and different geographic distribution of local HALE compared with mortality measures, along with the additional benefit of being able to model the complex interaction of mortality and morbidity, suggest that HALE may be a useful population health measure. PMID- 11007661 TI - [ [In Process Citation] PMID- 11007659 TI - Ontario familial colon cancer registry: methods and first-year response rates. AB - The Ontario Familial Colon Cancer Registry (OFCCR) is a novel registry that collects family history information, epidemiologic data, blood samples and tumour specimens from a population-based sample of colorectal cancer patients and their families. Families are classified as either high familial risk, intermediate familial/other risk or low (sporadic) risk for colorectal cancer. Obtaining high response rates in genetic family studies is especially challenging because of both the time commitment required and issues of confidentiality. The first-year response rate was 61%, resulting in 1,395 participating probands. In an attempt to assess potential response bias, we compared participants with non participants. The age and sex of participants did not differ from non participating probands; however, cases in rural areas were somewhat more likely to participate. To date, 57% of 1,587 relatives participated; females were more likely to participate, and relatives of low familial risk were least likely to participate. The OFCCR is an excellent resource that will facilitate the study of genetic and environmental factors associated with colorectal cancer. PMID- 11007662 TI - [Thrombus remodeling. Key factor in the progression of coronary atherosclerosis]. AB - The mechanism of the progression of atherosclerosis involves the rupture of a plaque with thrombus formation, followed by an invasion of monocytes that induce the formation of tissue factor and a second thrombus. This is followed by activation of the smooth muscle cells of the arterial wall with formation of connective tissue that infiltrates both thrombi. We can try to prevent atherosclerosis progression by inhibiting thrombosis with an inhibitor of tissue factor, inhibiting the process of scarring with Rapamycin, or both. Nowadays, we can study these processes using highly sensitive techniques to monitorize the formation and evolution of the thrombus, and magnetic resonance imaging that allow us to study the growth of the connective tissue. With these techniques we can study the natural history of atherosclerosis and the efficacy of different drugs to prevent its progression. PMID- 11007663 TI - [Biochemical diagnosis of hypertensive myocardial fibrosis]. AB - A substantial increase in fibrillar collagen has been observed in the left cardiac ventricle of animals and humans with arterial hypertension. Hypertensive myocardial fibrosis is the result of both increased collagen types I and III due to the fact that its synthesis by fibroblasts and myofibroblasts is stimulated and its extracellular collagen degradation unchanged or decreased extracellular collagen degradation. Hemodynamic and non-hemodynamic factors may be involved in the disequilibrium between collagen synthesis and degradation that occurs in hypertension. As shown experimentally and clinically, an exaggerated rise in fibrillar collagen content promotes abnormalities of cardiac function, contributes to the decrease in coronary reserve and facilitates alterations in the electrical activity of the left ventricle. Although microscopic examination of cardiac biopsies is the most reliable method for documenting and measuring myocardial fibrosis, the development of non-invasive methods to indicate the presence of myocardial fibrosis in hypertensive patients would be useful. We have therefore applied a biochemical method based on the measurement of serum peptides derived from the tissue formation when synthesized and degradation of fibrillar collagens to monitor the turnover of these molecules in rats with spontaneous hypertension and patients with essential hypertension. PMID- 11007664 TI - [Heart failure: the electrophysiological connection. Myocardial stunning and heart failure: mechanisms in common?]. AB - The delayed recovery of function after brief episodes of ischemia is known as stunning. Myocardial stunning and heart failure would, at first glance, appear to have little in common other than the obvious contractile dysfunction in both settings. Here I describe studies which shed new light on the underlying mechanisms of these two forms of contractile dysfunction, revealing unexpected fundamental similarities. PMID- 11007665 TI - [Dilated cardiomyopathy: recent advances and current treatment]. AB - The combination of dilatation and systolic dysfunction of the left or both ventricles from idiopathic or specific origin define dilated cardiomyopathy (DC). It is an important cause of cardiac morbidity through congestive heart failure (CHF) or arrhythmias. Prevalence studies estimate a rate of left ventricular systolic dysfunction of 2% to 3% or more, and of 1.5% of CHF among the general population. Genetic studies on familial DC have identified at least 5 genetic locus. In addition, the role of virus, genetic abnormalities, immunologic responses and increased myocardial apoptosis are factors recognized factors that play a significant role in the pathogenesis of idiopathic DC. Mortality in severe CHF may reach 50% at 2 years after diagnosis. The introduction of "triple" therapy combining diuretics, digoxin and angiotensin converting enzyme inhibitors (ACEI) has significantly decreased this high mortality. Recent large, multicentric clinical trials on drugs aimed to diminish the neuroendocrine hyperactivity of patients in stable chronic CHF (betablockers and spirolactone) have resulted in an additional reduction in total mortality of about 35% in relation with control groups. Betablockers, compared to ACEI, also diminish the rate of sudden death prompting their almost mandatory indication in the absence of contraindications. Cardiac transplantation presently offers a survival rate of 66% at 5 years but donor scarcity has stabilized the number of procedures. In patients resuscitated from malignant arrhytmias the implantation of defibrillators offers a better survival than drugs. Future advances in the knowledge of the pathogenesis and especially of genetic mechanisms, may substantially change the understanding and treatment of these disorders. PMID- 11007666 TI - [Aortic valve replacement with pulmonary autograft (the Ross procedure) in adult and pediatric patients]. AB - Aortic valve replacement with pulmonary autograft was first performed by Donald Ross in 1967. Initially, the procedure was not widely accepted, by Cardiologists and Cardiac surgeons fundamentally due to its complexity and demanding surgical technique, and because innumerous series two cardiac valves were at risk. The results published in the last 10-15 years established the pulmonary autograft as one of the best methods of aortic valve replacement, especially in pediatric patients and young adults. In the present article, we reviewed present indications and contraindications, and our clinical experience with 26 patients (pediatrics and adults). Analysis of the first 22 the patients with a minimum of 6 months of follow-up (180-620 days) was performed. Follow-up is complete (100%). Mean age was 31.4 +/- 12.6 years. Five patients were pediatrics (<= 14 years). Three patients (11%) with previous percutaneous procedures and 4 patients (14%) with previous surgical procedures. There was no early or late mortality. In the last follow-up, 19 of 22 (86.36%) had no autograft insufficiency (>= grade 1), and in one patient it was moderate (grade 2). The 2 remaining patients developed severe autograft insufficiency (grade 4) and were reoperated on, with satisfactory postoperative outcome. Mean maximal gradient was 7.85 +/- 5 mmHg at 18 months (3-29). Patients with preoperative aortic stenosis showed a significant reduction in myocardial mass index (208.7 +/- 32 a 95.8 +/- 28.8 g/m2) at 18 months. In these patients, septal and posterior wall thickness decreased significantly, in the first month. Two pediatric patients have developed transpulmonar gradient > 50 mmHg. One of them underwent successful stent implantation. We have not observed significant homograft insufficiency in any of our patients. All our patients remain asymptomatic (functional class I) without medical treatment. We have not observed either thromboembolic or haemorrhagic episodes, nor endocarditis. No patient is receiving anticoagulants. Clinical and echocardiographic mid term results in pulmonary autograft and homograft in our series, are excellent after the Ross procedure. PMID- 11007667 TI - [The heart transplantation in Spain. Organization and results]. AB - Heart transplantation has achieved a great development in Spain. With 8 heart transplantation procedures per million of inhabitants per year, we are the second ranking country, after the U.S.A., in this therapeutic activity. A relative adequacy between the number of available donors and the number of receptors included on the waiting list help to reduce the delays and the mortality rates in expecting patients, compare to other countries, which demonstrated the efficiency of the system. From 1984 to 1998, 2,756 heart transplantations were performed in Spain, in 16 authorized hospitals. In 56 cases (2%) the procedures were re transplantation. Cardiomyopathy with ischemic origin was the most frequent pathology in determining the intervention, affecting to the 37% of the receptors, followed by idiopathic dilated cardiomyopathy with a 35%. Compared to other countries, a high proportion of cardiopathies (11%) of rheumatic origin stands out.20% of the heart transplantations were performed in a maximum emergency situation of the receptors. Assigning donors priority on a national scale. In a 2, 9% of the cases, extra procedures of circulatory assistance devises were used. The actuarial survival of the whole series is 74% at the end of the first year; 62% after five years and 46% after ten years. The mean survival of the grafts was 9.5 years. These data do not differ substantially from those of the Registry of the International Society of Heart and Lung Transplantation. The most important causes of early mortality were the graft primary failure, the infections and the rejection. Among the causes of late morbidity, cancer, with a prevalence of 6,1%, and vascular disease of the graft, present in 8,4% of the receptors, stand out. These results support the so-called Spanish Model for Solid Organ Transplantation, which, according to the health authorities of the European Union, should be implemented in all the member countries. The keys to the success of the Spanish Model are based on three factors, in order of importance: a) to have a public professionalized institution for the detection of donors, the allocation of the grafts, and the coordination of the extraction and transportation force. The Transplant National Organization is the name of this complex force in our country; b) the high rates of road and labour mortality in Spain, and c) to provide incentives for the professionals in charge of the whole process. PMID- 11007668 TI - Ethical selection of living kidney donors. AB - Renal transplant centers vary markedly in their rates of living donor kidney transplantation. Recent surveys also document marked differences among centers in both appreciation of medical risk for donation and what constitutes ethical donor selection. Because of this marked variability, some donors likely are being inappropriately denied or others are being inappropriately accepted. In addition to defensible donor education about risk and benefit, three fundamental obligations of the center are identified: (1) to recognize that it is often ethical to participate in acts of individual risk and sacrifice that are performed to benefit others; (2) to not deny transplantation without good reason to donors and recipients who apply to the center; and (3) to neutralize, but not overreact to, center self-interest, which stems from the professional benefits of transplantation and the center's desire to help potential transplant recipients. The basic medical facts surrounding donation must be understood by all parties as part of ethical decision making. Donor risk can be presented quantitatively using US Renal Data System data as a baseline. Confirmation of accurate donor understanding of risks, benefits, and alternatives is always a fundamental center obligation. Donors should not be rejected except for the general reasons we identify, and when these reasons do not seem to apply, the decision to deny transplantation should be reconsidered. PMID- 11007669 TI - Soluble Fas and soluble Fas-ligand in children with Escherichia coli O157:H7 associated hemolytic uremic syndrome. AB - We measured soluble Fas-ligand (sFas-L) and soluble Fas (sFas) levels by sandwich enzyme-linked immunosorbeny assay and compared them among (1) healthy controls (n = 11), (2) children with hemorrhagic colitis (HC) caused by a non-verotoxin producing pathogen (n = 23), (3) patients with uncomplicated Escherichia coli O157:H7 HC (n = 14), and (4) children with O157:H7-associated hemolytic uremic syndrome (HUS) (n = 24). Children with uncomplicated E coli O157:H7 HC and HUS were matched for duration of enteric prodrome before blood sample collection. We also compared sFas-L and sFas levels among patients with HUS according to severity of renal dysfunction; abnormally increased sFas-L levels were noted in only 4% of the children (n = 3). Abnormally high concentrations of sFas were noted in 9% of the children with HC caused by a non-verotoxin-producing pathogen, 29% of the patients with uncomplicated E coli O157:H7 HC, and 69% of the children with O157:H7-associated HUS. Compared with healthy controls, patients with HUS had twofold greater concentrations of sFas (P: < 0.0001). Levels of sFas were not statistically different between 14 patients with uncomplicated O157:H7 HC and 14 children with HUS (8.2 +/- 4.7 versus 11.0 +/- 4.6 U/mL, respectively; P: < 0.07) when matched for time after onset of enteritis (7.0 +/- 3.7 versus 7.3 +/- 3.8 days, respectively). Greater concentrations of sFas were noted in patients with HUS who developed oligoanuria (n = 10; P: < 0.007), required peritoneal dialysis (n = 10; P: < 0.007), or had a decreased glomerular filtration rate (n = 5; P: < 0.002) 1 year later. Our data show that plasma concentrations of sFas but not sFas-L are abnormally increased in children with O157:H7 infections. Levels of sFas are associated with severity of renal dysfunction during HUS. Further studies are needed to clearly determine the role and origin of circulating sFas among children with infections caused by E coli O157:H7. PMID- 11007670 TI - A novel interpretation of the role of von Willebrand factor in thrombotic microangiopathies based on platelet adhesion studies at high shear rate flow. AB - Clinical manifestations of thrombotic microangiopathies (TMA) are secondary to platelet aggregation and thrombotic occlusion of the microvasculature of the affected organs. Abnormalities in von Willebrand factor (vWF) in these patients were considered instrumental in promoting the process leading to microvascular thrombosis. We evaluated the capacity of plasma in these patients to induce adhesion of normal platelets and thrombus formation under conditions of controlled fluid shear stress. We also studied vWF multimeric distribution to establish whether abnormalities of this glycoprotein correlate with platelet adhesion and thrombus formation. Plasma from patients in the acute phase and remission showed the same capacity to induce platelet adhesion and thrombus formation at a low level of shear rate (600 sec(-1)) as plasma from control subjects. At a high shear rate (1,500 sec(-1)), platelet adhesion and thrombus dimensions were significantly increased (P: < 0.05) by plasma from patients with TMA compared with controls. The capacity to enhance thrombus formation at high shear stress was present during the acute phase and disease remission and did not correlate with the presence of unusually large vWF multimers. Increased thrombus formation with patient plasma is completely normalized by blocking the interaction of vWF with the platelet receptors, glycoprotein (GP)Ib and GPIIb IIIa, suggesting that the phenomenon is completely mediated by vWF. Our results suggest the possibility of an intrinsically altered vWF molecule in these patients that is probably more effective than normal vWF in mediating platelet adhesion and thrombus formation. PMID- 11007671 TI - Characterization of early IgA nephropathy. AB - Histological grading of 45 patients with clinical early immunoglobulin A (IgA) nephropathy was correlated with disease progression over a median follow-up of 123 months. Clinical early IgA nephropathy was defined as a serum creatinine level of 1.3 mg/dL or less, proteinuria of 0.4 g/d or less of protein, and the absence of hypertension at the time of renal biopsy. Disease progression was related to the occurrence of impaired renal function, increased proteinuria, and hypertension. We applied a previously described chronicity-based histological grading to the renal biopsy specimen and also assessed acute glomerular lesions. Disease progression was observed in 44.4% of these patients. Forty patients (89%) showed glomerular grade 1 (GG1) and 5 patients (11%) showed GG2, but this grading did not correlate with disease progression. However, when GG1 was subdivided into GG1a (mean sclerosis per glomerulus <10%) and GG1b (mean sclerosis per glomerulus 10% to <25%), GG1a correlated with nonprogressive disease. Tubulointerstitial grade also correlated with disease progression but was associated with a low sensitivity for predicting nonprogressive disease. Hyaline arteriolosclerosis and acute glomerular lesions did not correlate with disease progression. The chronicity-based histological grading is not only applicable to clinical early IgA nephropathy, but also more importantly, it characterizes GG1a in a subset of patients with a very low risk for disease progression, which can be regarded as genuine early IgA nephropathy. PMID- 11007672 TI - ANCA-associated crescentic glomerulonephritis with mesangial IgA deposits. AB - Antineutrophil cytoplasmic autoantibodies (ANCA) are commonly associated with a necrotizing and crescentic glomerulonephritis (GN) that is pauci-immune, with few or no glomerular immune complex deposits detectable by immunofluorescence (IF) or electron microscopy (EM). Immunoglobulin A (IgA) nephropathy may also be manifest as a crescentic GN, but it is characterized by mesangial immune complex deposits containing IgA and is rarely associated with myeloperoxidase (MPO)- or proteinase 3 (PR3)-specific ANCA when an enzyme immunoassay is used to detect these antibodies. This report describes six patients with severe crescentic GN with mesangial IgA deposits by IF and mesangial electron-dense deposits by EM in patients with positive ANCA serological test results (four patients, anti-PR3; one patient, anti-MPO; one patient, anti-PR3 and anti-MPO). Patients presented with acute or progressive renal insufficiency, hematuria, proteinuria (nephrotic range in two patients), and hypertension. Three patients had evidence of systemic vasculitis: two patients at initial presentation and one patient later in the clinical course. Renal biopsy specimens showed crescents in greater than 50% of glomeruli in all cases, but only mild, focal and segmental mesangial and endocapillary hypercellularity, more typical of ANCA-associated crescentic GN than of crescentic IgA nephropathy without associated ANCA. Semiquantitative analysis of mesangial and endocapillary cellularity performed on renal biopsy slides from these six patients and from eight ANCA-negative patients with IgA nephropathy and crescents in greater than 50% of glomeruli showed significantly greater hypercellularity in the ANCA-negative cases. Three of five ANCA-positive patients for whom follow-up clinical data were available showed improved renal function after treatment with cyclophosphamide and corticosteroids and have not developed end-stage renal disease 17, 20, and 25 months postbiopsy. The remaining two patients were dialysis dependent at the time of biopsy and have remained so despite treatment with cyclophosphamide and corticosteroids. The findings suggest an overlap syndrome of ANCA-associated crescentic GN and IgA nephropathy that resembles the former both histologically and in its potential to respond to aggressive therapy if detected relatively early in its course. PMID- 11007673 TI - Effect of posture on sodium excretion and diuretic efficacy in nephrotic patients. AB - It is well known that posture affects natriuresis in cirrhosis and heart failure. This study evaluates the role of posture on spontaneous urinary salt excretion (U(Na)V) and diuretic-induced natriuresis in nephrotic patients with mild renal impairment. U(Na)V and plasma concentrations of the main hormones involved in sodium regulation were evaluated at baseline (Baseline) and after furosemide administration (20 mg intravenously at 8:00 AM [Diuretic]) in seven nephrotic patients with mild renal impairment (creatinine clearance, 68.5 +/- 7.6 mL/min) in either the supine or upright position for 6 hours (from 8:00 AM to 2:00 PM). At baseline, U(Na)V was greater in the supine than upright position (sodium, 51.8 +/- 6.2 versus 38.3 +/- 6.1 mEq/d; P: < 0.01). Similarly, furosemide was more effective in increasing U(Na)V in the supine (sodium, 51.8 +/- 6.2 to 87.4 +/- 9.1 mEq/d; P: < 0.005) than upright position (sodium, 38.3 +/- 6.1 to 59.0 +/- 6.8 mEq/d; P: = not significant). Consequently, body weight decreased in the supine but not the upright position (-0.73 +/- 0.15 versus -0.17 +/- 0.22 kg; P: < 0. 05). Peripheral renin activity (PRA) and plasma aldosterone (Aldo) concentrations were greater in the upright than supine position at both Baseline and Diuretic. A similar pattern was observed for hematocrit, used as an index of plasma volume. In addition, a positive correlation was detected between hematocrit and PRA (r = 0.89; P: < 0.001) in the upright position. Postural changes did not influence plasma concentrations of atrial natriuretic peptide. These data indicate that in nephrotic patients with mild impairment of glomerular filtration rate, the upright position causes a reduction in plasma volume; this hypovolemia activates the renin-Aldo system responsible for sodium retention in unstimulated conditions and a blunted natriuretic response to furosemide. PMID- 11007674 TI - Increased prevalence of polycystic kidney disease type 2 among elderly polycystic patients. AB - Autosomal dominant polycystic kidney disease (ADPKD) is genetically heterogeneous, with at least three chromosomal loci (PKD1, PKD2, and PKD3) accounting for the disease. Mutations in the PKD2 gene, on the long arm of chromosome 4, are estimated to be responsible for 15% of the cases of ADPKD, based on linkage studies. PKD2 is a milder form of the disease, with a mean age of end-stage renal disease (ESRD) approximately 20 years later than PKD1. The object of this study is to determine the proportion of elderly patients with ADPKD with ESRD who harbor mutations in the PKD2 gene. We analyzed all exons and intron-exon boundaries of the PKD2 gene by single-strand conformation polymorphism analysis and silver staining technique in 46 patients with ADPKD who reached ESRD after the age of 63 years or were not yet undergoing renal replacement therapy (RRT) by that age. We performed exactly the same studies in a control group of 40 patients with ADPKD with unknown gene status aged younger than 63 years. In 22 patients, a mutation in the PKD2 gene was defined: 18 of 46 patients from the elderly group and 4 of 40 patients from the control group. We identified 14 different mutations: 4 nonsense mutations, 1 missense mutation, 5 small deletions, 2 insertions, 1 deletion of the whole PKD2 gene, and 1 splicing mutation. Five of these mutations previously were described by our group. Three of the mutations reported in the present study are recurrent. The prevalence of PKD2 disease among elderly patients with ADPKD undergoing RRT is 39.1%, almost three times the prevalence of the disease in the general ADPKD population. PMID- 11007675 TI - Cytotoxicity of myeloma light chains in cultured human kidney proximal tubule cells. AB - We evaluated the effect of eight species of light chains on cultured human kidney proximal tubule cell proliferation. Exposure to light chains for 48 hours caused dose-dependent inhibition in tritium ((3)H)-thymidine incorporation by simian virus 40 immortalized human proximal tubule cells, although the effect was variable among different species of light chains. We studied cytotoxic effects of selected toxic light chains in further detail. Two of these light chains caused significant DNA degradation. A lambda-light chain caused lactate dehydrogenase release from exposed cells at 48 hours, but not at 24 hours. Cytomorphological and electron microscopic examination of cells exposed to light chains for 24 hours showed condensed nuclei, cell detachment, paucity of mitotic activity, and apoptosis, and at 48 hours of exposure, changes consistent with necrosis. Apoptosis assay by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling method showed a sixfold increase in the number of apoptotic cells exposed to the same lambda-light chain for 24 hours. Rhodamine phalloidin staining showed variable but significant disruptions in the actin cytoskeleton. These studies show that some myeloma light chains are toxic to cultured human proximal tubule cells and induce cytoskeletal injury and DNA damage consistent with apoptosis followed by secondary necrosis. Direct proximal tubule cell toxicity may be an important mechanism of renal involvement in multiple myeloma. PMID- 11007676 TI - Evidence that chronicity of hyponatremia contributes to the high urate clearance observed in the syndrome of inappropriate antidiuretic hormone secretion. AB - The high fractional excretion (FE) of uric acid observed in hyponatremia associated with the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) is commonly attributed to the volume-expanded state, although volume expansion in normonatremic volunteers is unable to increase urate clearance to a degree similar to that in SIADH. The goal of the present study is to analyze whether hyponatremia by itself could influence the FE of uric acid, as well as the effects of intravascular volume and glomerular filtration rate on FE of uric acid in SIADH. This study examines the effects of a 2-L infusion of isotonic saline over 24 hours on FE of uric acid in 9 normonatremic volunteers and 17 hyponatremic patients with SIADH. We also studied the FE of uric acid in 6 patients with SIADH with only mild water retention and the urate and creatinine clearances in 18 hyponatremic patients with SIADH before and after normalization of serum sodium levels by water restriction. When infusing 2 L of isotonic saline over 24 hours in healthy subjects, there was a decrease in plasma protein concentration of 8%, suggesting a similar degree of volume expansion than in patients with SIADH. The FE of uric acid did not increase to the same extent (9% +/- 1.5% versus 17% +/- 1.5%; P: < 0.01). Conversely, in 6 hyponatremic patients with mild water retention (1 L), the FE of uric acid was still high despite indirect signs of only a small increase in plasma volume. The mainstay of these observations is that chronicity of hyponatremia by itself could affect urate excretion. We also observed that in the patients with SIADH, high FE of uric acid inversely correlated with glomerular filtration rate (r = -0.66; P: < 0.01) only during the hyponatremic state. These data suggest that hyponatremia by itself, combined with mild volume expansion and glomerular filtration rate, has a role in the high FE of uric acid in the SIADH. PMID- 11007677 TI - Revascularization of renal artery stenosis in patients with renal insufficiency. AB - The incidence and prevalence of end-stage renal disease (ESRD), particularly in the elderly population, have continued to increase in the United States. It is estimated that 10% to 20% of the elderly patients with ESRD have potentially remediable renal vascular disease. The purpose of the present study is to examine the results of renal artery revascularization in 20 patients aged older than 55 years with chronic renal failure (serum creatinine level >2 mg/dL) with proximal renal artery stenosis (RAS) diagnosed by magnetic resonance angiography (MRA) who underwent surgical or percutaneous revascularization. Patients were followed up closely in the postrevascularization period; renal function was monitored and potential complications of the procedure were carefully noted. Four of the 20 patients developed serious complications, including 3 patients with clinically significant atheroembolic disease and 1 patient with renal artery dissection. Seven patients developed greater than 5% eosinophilia. Five of the 20 patients had a deterioration in renal function 3 to 6 months after the procedure, and only 5 patients had a reduction in serum creatinine concentration 3 to 6 months after the procedure. The present study suggests that in elderly patients with chronic renal failure and proximal RAS, revascularization of renal vessels is associated with a high complication rate, and improvement in renal function occurs in only 25% of the patients. Whether revascularization can slow the rate of progression of renal failure remains uncertain and can only be answered by a large prospective trial. PMID- 11007678 TI - Renal cell carcinoma in acquired cystic kidney disease: volume growth rate determined by helical computed tomography. AB - The aim of this study is to determine the growth rate and behavior of renal cell carcinoma in chronic hemodialysis patients with acquired cystic kidney disease (ACKD). Renal cell carcinomas in 17 hemodialysis patients (mean age, 52 +/- 11 years; mean hemodialysis duration, 7.2 +/- 3.3 years) with ACKD were examined with helical computed tomography (CT) for 0.5 to 6.0 years (mean, 2.1 +/- 1.9 years). The 17 renal cell carcinomas were histologically proven and graded after nephrectomy (16 patients) or autopsy (1 patient). Tumor volume was estimated by counting the number of pixels in the tumor and a 1-cm(2) area on helical computed tomographic scan using a personal computer. Estimated volume growth rates and doubling times of the carcinoma were correlated with histological grades. Fifteen of the 17 neoplasms (88%) were less than 3 cm in diameter at initial CT. The overall volume growth rate was 0.07 to 17.34 cm(3)/y (mean, 4.14 +/- 5.66 cm(3)/y), and the estimated volume-doubling time was 0. 08 to 23.31 years (mean, 5.09 +/- 6.99 years). The mean growth rate of the 3 grade 3 carcinomas was 6.01 +/- 4.50 cm(3)/y (range, 0.88 to 9.28 cm(3)/y), which was significantly greater than that of the 9 grade 1 carcinomas (0.40 +/- 0.40 cm(3)/y; range, 0.09 to 1.37 cm(3)/y) or the 5 grade 2 tumors (0.79 +/- 0.74 cm(3)/y; range, 0.12 to 2.00 cm(3)/y). Eleven of the 17 carcinomas (65%) had more than a 1-year volume doubling time. The 3 grade 3 neoplasms and 1 of the grade 2 lesions had less than a 0.5-year doubling time. PMID- 11007679 TI - Atrial natriuretic factor in oliguric acute renal failure. Anaritide Acute Renal Failure Study Group. AB - Atrial natriuretic peptide (ANP), an endogenous hormone synthesized by the cardiac atria, has been shown to improve renal function in multiple animal models of acute renal failure. In a recent multicenter clinical trial of 504 patients with acute tubular necrosis (oliguric and nonoliguric), ANP decreased the need for dialysis only in the oliguric patients. In the present study, 222 patients with oliguric acute renal failure were enrolled into a multicenter, randomized, double-blind, placebo-controlled trial designed to assess prospectively the safety and efficacy of ANP compared with placebo. Subjects were randomized to treatment with a 24-hour infusion of ANP (anaritide, 0.2 microgram/kg/min; synthetic form of human ANP) or placebo. Dialysis and mortality status were followed up for 60 days. The primary efficacy end point was dialysis-free survival through day 21. Dialysis-free survival rates were 21% in the ANP group and 15% in the placebo group (P = 0.22). By day 14 of the study, 64% and 77% of the ANP and placebo groups had undergone dialysis, respectively (P = 0.054), and 9 additional patients (7 patients, ANP group; 2 patients, placebo group) needed dialysis but did not receive it. Although a trend was present, there was no statistically significant beneficial effect of ANP in dialysis-free survival or reduction in dialysis in these subjects with oliguric acute renal failure. Mortality rates through day 60 were 60% versus 56% in the ANP and placebo groups, respectively (P = 0.541). One hundred two of 108 (95%) versus 63 of 114 (55%) patients in the ANP and placebo groups had systolic blood pressures less than 90 mm Hg during the study-drug infusion (P < 0.001). The maximal absolute decrease in systolic blood pressure was significantly greater in the anaritide group than placebo group (33.6 versus 23.9 mm Hg; P < 0.001). This well-characterized population with oliguric acute renal failure had an overall high morbidity and mortality. PMID- 11007680 TI - Intravenous iron dextran treatment in predialysis patients with chronic renal failure. AB - Iron deficiency anemia is common in patients with chronic renal failure not undergoing hemodialysis. Current therapy consists of oral or intravenous (IV) iron dextran (IVID). The standard IV regimen is 100 to 200 mg/dose for a 1-g total dose. We hypothesized that 500 mg/wk of IVID for two doses would be less costly and equally effective as 200 mg/wk for five doses. We prospectively studied 22 patients with creatinine clearances less than 50 mL/min who were not undergoing dialysis and had anemia and evidence of iron deficiency (ferritin level <100 ng/mL or transferrin saturation [TSAT] <20%). Patients were randomized into two groups: group I (n = 8), 200 mg/wk of IVID for 5 weeks, and group II (n = 14), 500 mg/wk of IVID for 2 weeks. All patients tolerated IVID infusions without serious adverse reactions. Over the 6-month follow-up, both groups experienced an increase in hemoglobin levels from baseline. Ferritin levels in both groups increased (P < 0.005), peaked at 2 weeks, then declined thereafter. Over the 6-month follow-up, both groups experienced significant improvement, although the beneficial effects of group II declined at a significantly faster rate than group I (P = 0.003). There was no significant difference in change in ferritin levels between groups. TSAT peaked at 2 weeks in both groups (P < 0. 001). Group I experienced a significant increase in TSAT throughout the 6-month follow-up (P < 0.03), and group II achieved a significant increase in TSAT at 2 weeks, but not at 3 and 6 months. There was no significant difference in pretreatment to posttreatment change in TSAT. Treatment in group II was 35.2% more cost-effective than in group I ($965 versus $1,490, respectively). We conclude that IVID, 500 mg/wk, for 2 weeks is as effective and safe as 200 mg/wk for 5 weeks, but much less costly. PMID- 11007681 TI - Prevalence of sleep disturbances in chinese patients with end-stage renal failure on continuous ambulatory peritoneal dialysis. AB - Patients with end-stage renal failure (ESRF) are reported to have a high prevalence of sleep disorders, such as daytime sleepiness, insomnia, restless legs syndrome (RLS), and obstructive sleep apnea syndrome (OSAS). However, there are few published data from Southeast Asia. A sleep questionnaire was administered to 201 patients (103 men) at the continuous ambulatory peritoneal dialysis (CAPD) outpatient clinic to assess sleep problems. Patients had a mean age of 56.7 +/- 12 (SD) years, with a mean body mass index (BMI) of 23.6 +/- 3.5 kg/m(2). Daytime sleepiness was the most frequent symptom (77.1%), and frequent awakening occurred in 69% of the patients. Sleep-onset insomnia and sleep maintenance insomnia occurred in 73% and 60% of the patients, respectively. Sixty two percent of the patients reported symptoms of RLS, which significantly correlated with sleep-onset insomnia (odds ratio [OR], 2.9; 95% confidence interval [CI], 1.5 to 5.5; P = 0.001) and sleep-maintenance insomnia (OR, 2.1; 95% CI, 1.2 to 3.8; P = 0.014). The prevalence of OSAS was estimated by the frequency of the following symptoms: extremely loud snoring, 7 patients (3.5%); observed choking, 21 patients (10.5%); witnessed apnea, 11 patients (5.6%); snoring and witnessed apnea, 6 patients (3%); disruptive snoring, 29 patients (14.4%); and disruptive snoring and witnessed apnea, 3 patients (1.5%). This questionnaire survey confirmed a high prevalence of daytime sleepiness, insomnia, and RLS in patients with ESRF undergoing CAPD but showed a relatively low prevalence of OSAS of up to 14.4%, which may be related to the low BMI of these patients with ESRF compared with other populations. Whether this contributes to the overall better survival observed in some Asian patients with ESRF undergoing dialysis needs further investigation. PMID- 11007682 TI - Effect of the vitamin D analogues paricalcitol and calcitriol on bone mineral in vitro. AB - Paricalcitol (19-nor-1,25-dihydroxyvitamin D(2)), a new vitamin D analogue, recently became available for the treatment of hyperparathyroidism in patients with end-stage renal disease. It is safe and effective in suppressing parathyroid hormone, with apparently less propensity for hypercalcemia than calcitriol (1, 25 dihydroxyvitamin D(3)). However, the mechanism of action on bone has not been fully elucidated. This study compares the effects of paricalcitol and calcitriol on the bone mineral. Neonatal (5- to 7-day-old) mouse calvariae were incubated in the absence or presence of either paricalcitol or calcitriol for 48 hours, and calcium flux, osteocalcin and acid and alkaline phosphatase activity, and interleukin-6 (IL-6) release were determined. Increasing concentrations of both calcitriol and paricalcitol increased calcium efflux. At lower concentrations, paricalcitol had no effect on acid phosphatase activity; however, at 10(-8) mol/L, paricalcitol caused a significant increase similar to that of calcitriol at 10(-9) mol/L. Increasing concentrations of paricalcitol had no effect on alkaline phosphatase activity, whereas calcitriol (10(-8) mol/L) caused significant inhibition. At low concentrations, paricalcitol had no effect on osteocalcin release; however, at 10(-8) mol/L, both compounds significantly increased osteocalcin production. Neither compound had an effect on IL-6 release. These data show that: (1) at low concentrations, both compounds induce a similar calcium efflux from cultured bone; (2) at low concentrations, paricalcitol has no effect on osteocalcin or acid and alkaline phosphatase activity; (3) at greater concentrations, paricalcitol and calcitriol have similar effects on acid phosphatase and osteocalcin activity; (4) calcitriol, but not paricalcitol, inhibits alkaline phosphatase release; and (5) the bone-resorbing effect of both compounds is independent of IL-6 release. Thus, although both compounds have similar effects on calcium efflux from bone, at therapeutic concentrations, paricalcitol does not seem to inhibit osteoblast activity. This may explain, in part, the lower calcemic effect of paricalcitol. PMID- 11007683 TI - Assessment of sonographic venous peak systolic velocity in detecting hemodialysis arteriovenous graft stenosis. AB - There is no single effective means of assessing arteriovenous access function, although monitoring hemodialysis venous pressure (VP) or measuring access recirculation may be of some benefit. The present study assesses prospectively the efficacy of following the peak systolic velocity (PSV) as a single measure to detect arteriovenous graft (AVG) stenosis. PSV was measured in 12 patients after new AVG placement and at approximately 2-month intervals. Angiography was also performed after new graft placement and when PSV was elevated to greater than 200 cm/sec, hemodialysis access VP increased to greater than 150 mm Hg on three consecutive readings, or access recirculation increased to greater than 11%. PSV was then compared with results from angiography, VP monitoring, and access recirculation. The 12 patients underwent 34 PSV studies, followed by angiography on 25 occasions. Each patient underwent at least one angiogram. Each abnormal PSV value was confirmed with the finding of stenosis on angiogram, except for two patients with PSVs greater than 400 cm/sec and normal angiography results. VP and recirculation were not elevated. During this period, two patients developed thrombosis of the AVG, and two patients underwent angioplasty with improvement in PSV. We conclude that elevations in PSV measured at the venous anastomosis are an effective means of screening for AVG stenosis, AVG stenosis can occur early after AVG placement, and elevated VP and recirculation are late findings in AVG dysfunction. PMID- 11007684 TI - Salvage of occluded autologous arteriovenous fistulae. AB - Well-functioning vascular access is essential to optimizing outcomes in hemodialysis patients. Initial placement of greater numbers of autologous arteriovenous fistulae (AVFs) is needed, as is more attention to salvage of such accesses when they become occluded. In this study, thrombolysis with small doses of thrombolytic agents in combination with balloon angioplasty was attempted in 15 patients with 16 autologous AVFs (7 forearm AVFs, 8 upper-arm AVFs). Either urokinase (UK; mean dose, 109,375 U) or tissue plasminogen activator (tPA; mean dose, 7.1 mg) was used for thrombolysis. The technical success rate was 94%, whereas long-term patency was achieved in 81% of the accesses. Failure to achieve long-term patency was caused by the inability to achieve adequate drainage in 2 patients. In 1 patient, technical inability to cannulate the access and penetrate the venous end of a basilic vein transposed fistula occurred. One patient was successfully declotted and angioplastied twice. Successful salvage of thrombosed AVFs can be safely performed with much lower doses of thrombolytic agents than previously reported. In addition, tPA is effective in this setting, in addition to UK. Salvage of occluded autologous AVFs should be attempted more frequently than is currently practiced and would result in improved hemodialysis patient outcomes. PMID- 11007685 TI - Anthropometric measures and risk of death in children with end-stage renal disease. AB - We evaluated the association between anthropometric measurements and death among pediatric patients with end-stage renal disease (ESRD) using data from the Pediatric Growth and Development Special Study (PGDSS) from the US Renal Data System. Height, growth velocity, and body mass index (BMI) were used for the analysis of 1,949 patients in the PGDSS. To standardize these measurements, SD scores (SDSs) were calculated using population data from the Third National Health and Nutrition Examination Survey. Using Cox proportional hazards models, we assessed the association between anthropometric measures and death, controlling for demographic factors and stratifying by age. Multivariate analysis showed that each decrease by 1 SDS in height was associated with a 14% increase in risk for death (adjusted relative risk [aRR], 1.14; 95% confidence interval [CI], 1.02 to 1.27; P = 0.017). For each 1 SDS decrease in growth velocity among patients in our sample, the risk for death increased by 12% (aRR, 1.12; 95% CI, 1.00 to 1.25; P = 0.043). There was a statistically significant U-shaped association between BMI and death (P = 0.001), with relatively low and high BMIs associated with an increased risk for death. In children with ESRD, growth delay and extremes in BMI are associated with an increased risk for mortality. PMID- 11007686 TI - Long-term prognosis and incidence of acute myocardial infarction in patients on chronic hemodialysis. The Okinawa Dialysis Study Group. AB - Mortality from cardiovascular disease is high in chronic dialysis patients. We observed the occurrence of acute myocardial infarction (AMI) in the chronic dialysis population in Okinawa, Japan. A total of 3,741 chronic dialysis patients (2,073 men, 1,668 women) were followed up for 10 years from April 1, 1988, to March 31, 1998. Only definite cases of AMI were registered. Data were compared with AMI registry data obtained from the general population of the same district. The total duration of observation was 15,748.8 patient-years. During the study period, 61 patients (40 men, 21 women) had AMI. The incidence of AMI was 3.9/1,000 patient-years (men, 4.4/1,000 patient-years; women, 3.1/1,000 patient years). Twenty-four percent of the AMI cases occurred at 12 months after starting dialysis therapy. Mean age at onset of AMI was 60.9 +/- 11. 4 (SD) years; 58.9 +/ 11.4 years in men and 64.7 +/- 10.7 years in women. Survival rates after AMI were 50.8% at 1 month, 45.0% at 6 months, 36.5% at 12 months, and 13.0% at 44 months. Patients with diabetes mellitus (DM) had a greater incidence of AMI and a worse prognosis than patients without DM. The long-term prognosis of AMI was poor in chronic dialysis patients. PMID- 11007687 TI - Effect of apolipoprotein E polymorphism on serum lipid, lipoproteins, and atherosclerosis in hemodialysis patients. AB - Atherosclerosis and cardiovascular disease are the main causes of death in hemodialysis patients. Possession of the apolipoprotein E4 (ApoE4) allele has been associated with increased levels of serum lipids and with coronary and carotid artery atherosclerosis. We investigated the possible relationship between ApoE polymorphism and atherosclerosis risk factors in hemodialysis patients. Two hundred sixty-nine hemodialysis patients (115 women, 154 men) were included in our study. The mean patient age and mean hemodialysis duration were 45.8 +/- 15.3 years and 52.6 +/- 40.6 months, respectively. Testing was done on all patients to determine ApoE genotype and serum levels of total cholesterol (T-Cho), low density lipoprotein (LDL-C), high-density cholesterol (HDL-C), triglyceride (TG), lipoprotein (a) (Lp[a]), intact parathormone (iPTH), and fibrinogen. ApoE genotype was identified with the polymerase chain reaction. Ultrasonographic measurement of carotid artery intima media thickness (IMT) was used to diagnose atherosclerosis. We also analyzed ApoE polymorphism and risk factors such as age, gender, duration of hemodialysis, smoking, and hypertension in relation to the presence of atherosclerosis. Serum T-Cho and LDL-C levels were higher in patients with the ApoE4/3 phenotype than in those with ApoE3/3 and ApoE3/2 phenotypes (P < 0.05). However, there was no statistically significant link between ApoE polymorphism and serum levels of TG, HDL-C, or Lp(a) (P > 0.05). Apart from a relationship with age and duration of hemodialysis (P < 0.05), we found no significant association between atherosclerosis and ApoE polymorphism or the other risk factors analyzed (P > 0.05). In conclusion, although ApoE polymorphism significantly affects serum levels of T-Cho and LDL-C in hemodialysis patients, this study indicates that ApoE polymorphism is not associated with the presence of atherosclerosis in these individuals. The high incidence of atherosclerosis in these patients underlines the need for further research on other possible causative factors. PMID- 11007688 TI - Renal arterial intervention and angiotensin blockade in atherosclerotic nephropathy. AB - Atherosclerotic renal arterial disease (ARAD) is becoming a more important cause of end-stage renal failure. Diagnosis is more easily achieved because of greater clinical suspicion and more refined screening tools. However, the medical and interventional management of patients with ARAD is not well defined in the literature because there have been few randomized trials. Because the use of angiotensin-converting enzymes (ACE) inhibitors, and more recently angiotensin antagonists, has become much more widespread, it is inevitable that we should, knowingly or not, give these drugs to patients with ARAD. We describe 2 case studies in which the angiotensin-antagonist irbesartan was given to 2 patients with effectively single-functional kidneys after successful renal arterial radiologic intervention. The rationale for the use of irbesartan was to control BP, which had not responded to the initial arterial intervention, and took place in patients both refractory to, and intolerant of, many other anti-hypertensive drugs. Irbesartan successfully and safely reduced systemic BP, measured by use of ambulatory BP, without prejudicing renal function (measured by use of individual kidney function GFR). PMID- 11007689 TI - Cyclosporine and tacrolimus-associated thrombotic microangiopathy. AB - The development of thrombotic microangiopathy (TMA) associated with the use of cyclosporine has been well documented. Treatments have included discontinuation or reduction of cyclosporine dose with or without concurrent plasma exchange, plasma infusion, anticoagulation, and intravenous immunoglobulin G infusion. However, for recipients of organ transplantation, removing the inciting agent is not without the attendant risk of precipitating acute rejection and graft loss. The last decade has seen the emergence of tacrolimus as a potent immunosuppressive agent with mechanisms of action virtually identical to those of cyclosporine. As a result, switching to tacrolimus has been reported to be a viable therapeutic option in the setting of cyclosporine-induced TMA. With the more widespread application of tacrolimus in organ transplantation, tacrolimus associated TMA has also been recognized. However, literature regarding the incidence of the recurrence of TMA in patients exposed sequentially to cyclosporine and tacrolimus is limited. We report a case of a living donor renal transplant recipient who developed cyclosporine-induced TMA that responded to the withdrawal of cyclosporine in conjunction with plasmapheresis and fresh frozen plasma replacement therapy. Introduction of tacrolimus as an alternative immunosuppressive agent resulted in the recurrence of TMA and the subsequent loss of the renal allograft. Patients who are switched from cyclosporine to tacrolimus or vice versa should be closely monitored for the signs and symptoms of recurrent TMA. PMID- 11007690 TI - Endocapillary proliferative glomerulonephritis in a patient with parvovirus B19 infection. AB - A 45-year-old woman developed acute nephritic syndrome after erythema infectiosum. Laboratory data on admission showed decreased serum C3, C4, and CH50 levels and the presence of both immunoglobulin (Ig) M and IgG antibodies to human parvovirus B19 (HPV). A renal biopsy showed diffuse endocapillary proliferative glomerulonephritis. Immunofluorescence microscopy indicated 2+ granular staining for IgG, IgM, and C3 over the mesangial area and along glomerular capillary walls. HPV antigen was also detected in glomeruli by immunohistochemistry. Electron microscopy showed electron-dense deposits in the subendothelial space and the paramesangial area. These findings suggest that immune complex-type glomerulonephritis is caused by glomerular deposition of HPV antigen-antibody complexes in some patients with HPV infection. PMID- 11007691 TI - Choroidal calcifications in patients with Gitelman's syndrome. AB - Gitelman's syndrome is a renal tubular disorder characterized by a sodium and chloride reabsorption defect in distal tubular cells that determines hypokalemia, metabolic alkalosis, hypomagnesemia, and low calcium excretion. The presence of choroidal calcifications was sought in five patients with Gitelman's syndrome by ophthalmic examination, fluorescein angiography, indocyanine green angiography, and ocular ultrasonography. Calcifications observed in the choroid of two patients were shown by ultrasonography in both patients. Ophthalmic and fluorangiographic examinations detected this alteration in one of the two subjects. Chondrocalcinosis was found in one patient with choroidal calcifications. These findings suggest that precipitation of calcium salts can occur in the choroidal tissue of patients with Gitelman's syndrome. Deposits appeared to be well seen by ultrasonography because of their depth in ocular tissues. Sclerochoroidal calcifications may be favored by the low calcium excretion, which is associated with normal intestinal calcium absorption in patients with Gitelman's syndrome. PMID- 11007692 TI - Mediators, markers, and modulators in the hemolytic uremic syndrome. PMID- 11007693 TI - Toward more rational management of ischemic nephropathy: the need for clinical evidence. PMID- 11007694 TI - Understanding how aldosterone increases sodium transport. PMID- 11007695 TI - A 14-year-old boy with kidney allograft failure in the first month after transplantation. PMID- 11007696 TI - Ace inhibitors and erythropoietin response in hemodialysis patients. PMID- 11007697 TI - Indinavir nephropathy PMID- 11007698 TI - Continuing medical education exercise, october 2000 PMID- 11007699 TI - Pauci-immune ANCA-positive crescentic glomerulonephritis associated with metastatic adenocarcinoma of the lung. AB - A 66-year-old woman developed rapidly progressive renal failure several days after she was diagnosed with non-small cell carcinoma of the lung. Antineutrophil cytoplasmic antibody test performed as an indirect immunofluorescence assay was positive with a perinuclear pattern of staining (pANCA). The patient did not improve with hemodialysis treatment and died on the second day after admission to the hospital. A complete autopsy was performed and showed metastatic adenocarcinoma of the lung and pauci-immune crescentic glomerulonephritis. A literature search showed only 7 previously reported cases of malignant tumors associated with ANCA-positive pauci-immune crescentic glomerulonephritis. The clinicopathologic findings of the current and all previously reported cases and possible relationship between ANCA-positive glomerulonephritis and malignancy are discussed. PMID- 11007700 TI - Familial interstitial nephritis with progressive renal failure. AB - We describe a 53-year-old woman with chronic interstitial nephritis and asymptomatic impairment of renal function. Seven members of her family were suffering from renal failure and underwent hemodialysis. At the time of their hospital admissions, they had shown evidence of end-stage renal failure at 40 to 50 years of age. Lack of proteinuria, hematuria, hypertension, hyperuricemia, hearing loss, and visual impairment were present before the deterioration of the renal function. Renal biopsy of the presented case indicated chronic interstitial nephritis without glomerular basement membrane abnormalities. Progressive decline of renal function and the inheritance pattern of autosomal dominance in this family suggested the diagnosis of familial interstitial nephritis. PMID- 11007701 TI - Parathyroid carcinoma in a dialysis patient. AB - A 75-year-old woman who had been receiving dialysis for 3 years and had a long history of chronic renal failure attributable to reflux nephropathy was investigated for progressive hypercalcemia in the context of very high intact parathormone (iPTH) levels. Imaging showed two functional parathyroid glands in the neck. At parathyroidectomy, four variously enlarged parathyroid glands were found and completely resected, without autotransplantation. The histology of one of the glands showed invasive parathyroid carcinoma. Parathyroid carcinoma is a very rare condition, with only 16 previous cases involving dialysis patients described in the literature. We review the literature to draw together presentational and therapeutic information on the management of this problem in the setting of renal replacement therapy. PMID- 11007702 TI - Granulomatous tubulointerstitial nephritis in the renal allograft. AB - Granulomatous tubulointerstitial nephritis has rarely been described in renal allografts. Of 1,574 renal allograft tissue specimens obtained from 514 patients in the period 1993 to 1998, we report three cases (0.6%) with interstitial nephritis containing multiple noncaseating granulomas. Biopsy specimen 1 was obtained from a 44-year-old woman with a 6-day history of systemic Candida albicans infection and showed multiple granulomas containing budding yeasts. Biopsy specimen 2 was from a 33-year-old man who presented with miliary spread of Mycobacterium tuberculosis 12 days before the allograft biopsy. Biopsy specimen 3 was from a 23-year-old woman who presented with Escherichia coli urinary infection and bacteremia that was treated with antibiotics for 10 days before the biopsy. Granulomatous inflammation in reponse to infectious agents or drugs in immunosuppressed kidney transplant recipients can rarely give rise to allograft interstitial nephritis that is distinct from acute rejection. To our knowledge, there are no prior reports of granulomatous tubulointerstitial nephritis associated with C albicans and E coli infection or antibiotic therapy in human renal allografts. PMID- 11007703 TI - In pursuit of novel therapies based on an understanding of renal growth PMID- 11007704 TI - Adolescent depression: window of (missed?) opportunity. PMID- 11007705 TI - Genetic epidemiology of major depression: review and meta-analysis. AB - OBJECTIVE: The authors conducted a meta-analysis of relevant data from primary studies of the genetic epidemiology of major depression. METHOD: The authors searched MEDLINE and the reference lists of previous review articles to identify relevant primary studies. On the basis of a review of family, adoption, and twin studies that met specific inclusion criteria, the authors derived quantitative summary statistics. RESULTS: Five family studies met the inclusion criteria. The odds ratios for proband (subjects with major depression or comparison subjects) versus first-degree relative status (affected or unaffected with major depression) were homogeneous across the five studies (Mantel-Haenszel odds ratio=2.84, 95% CI=2.31-3.49). No adoption study met the inclusion criteria, but the results of two of the three reports were consistent with genetic influences on liability to major depression. Five twin studies met the inclusion criteria, and their statistical summation suggested that familial aggregation was due to additive genetic effects (point estimate of heritability of liability=37%, 95% CI=31%-42%), with a minimal contribution of environmental effects common to siblings (point estimate=0%, 95% CI=0%-5%), and substantial individual-specific environmental effects/measurement error (point estimate=63%, 95% CI=58%-67%). The literature suggests that recurrence best predicts the familial aggregation of major depression. CONCLUSIONS: Major depression is a familial disorder, and its familiality mostly or entirely results from genetic influences. Environmental influences specific to an individual are also etiologically significant. Major depression is a complex disorder that does not result from either genetic or environmental influences alone but rather from both. These findings are notably consistent across samples and methods and are likely to be generally applicable. PMID- 11007706 TI - Cost-effectiveness of services for mentally ill homeless people: the application of research to policy and practice. AB - OBJECTIVE: About one-quarter of homeless Americans have serious mental illnesses. This review synthesizes research findings on the cost-effectiveness of services for this population and their relevance for policy and practice. METHOD: Service interventions for seriously mentally ill homeless people were grouped into three overlapping categories: 1) outreach, 2) case management, and 3) housing placement and transition to mainstream services. Data were reviewed both from experimental studies with high internal validity and from observational studies, which better reflect typical community practice. RESULTS: In most studies, specialized interventions are associated with significantly improved outcomes, most consistently in the housing domain, but also in mental health status and quality of life. These programs are also associated with increased use of many types of health service and housing assistance, resulting in increased costs in most cases. The value of these programs to the public thus depends on whether their greater effectiveness is deemed to be worth their additional cost. CONCLUSIONS: Innovative programs for seriously mentally ill homeless people are effective and are also likely to increase costs in many cases. Their value ultimately depends on the moral and political value society places on caring for its least-well-off members. PMID- 11007707 TI - Neural networks: neural systems III. PMID- 11007708 TI - Respect from our medical colleagues: how much is too much? PMID- 11007709 TI - Naltrexone treatment for a 3-year-old boy with self-injurious behavior. PMID- 11007710 TI - William James, 1842-1910. PMID- 11007711 TI - Natural course of adolescent major depressive disorder in a community sample: predictors of recurrence in young adults. AB - OBJECTIVE: The primary purpose was to identify factors related to the recurrence of major depressive disorder during young adulthood (19-23 years of age) in a community sample of formerly depressed adolescents. METHOD: A total of 274 participants with adolescent-onset major depressive disorder were assessed twice during adolescence and again after their 24th birthday. Lifetime psychiatric information was obtained from their first-degree relatives. Adolescent predictor variables included demographic characteristics, psychosocial variables, characteristics of adolescent major depressive disorder, comorbidity, family history of major depressive disorder and nonmood disorder, and antisocial and borderline personality disorder symptoms. RESULTS: Low levels of excessive emotional reliance, a single episode of major depressive disorder in adolescence, low proportion of family members with recurrent major depressive disorder, low levels of antisocial and borderline personality disorder symptoms, and a positive attributional style (males only) independently predicted which formerly depressed adolescents would remain free of future psychopathology. Female gender, multiple major depressive disorder episodes in adolescence, higher proportion of family members with recurrent major depressive disorder, elevated borderline personality disorder symptoms, and conflict with parents (females only) independently predicted recurrent major depressive disorder. Comorbid anxiety and substance use disorders in adolescence and elevated antisocial personality disorder symptoms independently distinguished adolescents who developed recurrent major depressive disorder comorbid with nonmood disorder from those who developed pure major depressive disorder. CONCLUSIONS: Formerly depressed adolescents with the risk factors identified in this study are at elevated risk for recurrence of major depressive disorder during young adulthood and therefore warrant continued monitoring and preventive or prophylactic treatment. PMID- 11007712 TI - Identifying risk factors and key strategies in linkage to outpatient psychiatric care. AB - OBJECTIVE: The substantial failure of psychiatric patients to engage in outpatient specialty mental health care after an acute hospitalization at a time when managed care companies and others increasingly hold hospitals accountable for outcomes underscores the importance of identifying patients at high risk for not completing referrals. This study explored patient risk factors for not completing referrals and examined the success of several interventions targeted to achieving linkage with outpatient care. METHOD: A clinically detailed, structured form was used in abstracting information from the medical records of 229 inpatients with a primary psychiatric diagnosis. Clinicians and staff at outpatient programs were contacted to determine whether patients completed their referrals. RESULTS: Approximately two-thirds (65%) of the patients failed to attend scheduled or rescheduled initial outpatient mental health appointments after a hospital discharge. At high risk for unsuccessful linkage to outpatient care were patients with a persistent mental illness and those who had no prior public psychiatric hospitalization, were admitted involuntarily, and had longer lengths of stay. Controlling for risk factors, three clinical interventions used during the hospital stay more than tripled the odds of successful linkage to outpatient care: communication about patients' discharge plans between inpatient staff and outpatient clinicians, patients' starting outpatient programs before discharge, and family involvement during the hospital stay. CONCLUSIONS: Effective clinical bridging strategies can be used to avoid unnecessary gaps in the delivery of psychiatric services. Incorporating these strategies into routine care would enhance continuity of care, especially for some high-risk patients. PMID- 11007713 TI - Methods to improve diagnostic accuracy in a community mental health setting. AB - OBJECTIVE: This study determined the extent to which adding structured procedures improved diagnostic accuracy for outpatients with severe mental illness in a community mental health setting. METHOD: The Structured Clinical Interview for DSM-III-R (SCID) was used to interview 200 psychiatric outpatients. A research nurse reviewed medical records and amended the SCID diagnoses accordingly. A research psychiatrist or psychologist reviewed the diagnostic data and interviewed each patient to verify or further modify the previous findings. Diagnostic outcomes at each step of the procedure were compared to determine whether adding additional data improved diagnostic accuracy. The additional time required for each element of the diagnostic procedure was also assessed. RESULTS: Kappa comparisons of the different diagnostic levels showed that adding additional data significantly improved accuracy. Diagnoses rendered by combining the SCID and review of the medical record were the most accurate, followed by the SCID alone, and then diagnoses made by psychiatrists during routine care. In addition, the SCID alone identified five times as many current and past secondary diagnoses as were documented routinely in patients' charts. CONCLUSIONS: Combining structured interviewing with a review of the medical record appears to produce more accurate primary diagnoses and to identify more secondary diagnoses than routine clinical methods. The patients' knowledge of their diagnoses was limited, suggesting a need for patient education in this setting. Whether use of structured interviewing in routine practice improves patient outcomes deserves further study. PMID- 11007714 TI - Disability and quality of life in social phobia: epidemiologic findings. AB - OBJECTIVE: Social phobia in clinical studies is vividly associated with extensive disability and reduced quality of life. It is difficult to obtain a clear portrayal of the impairment associated with social phobia in community samples. Furthermore, it has been unclear in prior studies to what extent indices were attributable to social phobia as opposed to comorbid major depression. The authors examined relevant data from the Ontario Health Survey Mental Health Supplement. METHOD: The Ontario Health Survey Mental Health Supplement, a survey of more than 8,000 residents of Ontario, Canada, aged 15-64, used the University of Michigan Composite International Diagnostic Interview to assign DSM-III-R diagnoses. Several indicators of disability and quality of life were included. The authors compared these indices for persons with and without social phobia and adjusted where indicated for the effects of major depression and relevant sociodemographic factors. RESULTS: Persons with social phobia were impaired on a broad spectrum of measures, ranging from dropping out of school to experiencing disability in one's main activity. They were also significantly more likely than persons without social phobia to rate themselves as "low functioning" on the Quality of Well-Being Scale and to report dissatisfaction with many aspects of life. Depressive comorbidity seemed to contribute only modestly to these outcomes. CONCLUSIONS: Social phobia can be a serious, disabling anxiety disorder associated with marked reduction in quality of life. Impairment in social phobia is substantial, even in the absence of comorbid major depression. PMID- 11007715 TI - Anxiety in major depression: relationship to suicide attempts. AB - OBJECTIVE: This study was an examination of the relationship of lifetime panic disorder and anxiety symptoms at index hospitalization to a history of a suicide attempt in patients with a major depressive episode. METHOD: A total of 272 inpatients with at least one major depressive episode, with or without a history of a suicide attempt, were entered into the study. They were given structured diagnostic interviews for axis I and axis II disorders. Suicide attempt history, current psychopathology, and traits of aggression and impulsivity were also assessed. RESULTS: The rates of panic disorder did not differ in the suicide attempters and nonattempters. Agitation, psychic anxiety, and hypochondriasis were more severe in the nonattempter group. A multivariate analysis confirmed that this effect was independent of aggression and impulsivity. CONCLUSIONS: Comorbid panic disorder in patients with major depression does not seem to increase the risk for lifetime suicide attempt. The presence of greater anxiety in the nonattempters warrants further investigation. PMID- 11007716 TI - High vesicular monoamine transporter binding in asymptomatic bipolar I disorder: sex differences and cognitive correlates. AB - OBJECTIVE: It has been hypothesized that anomalies in monoaminergic function underlie some of the manifestations of bipolar disorder. In this study the authors examined the possibility that trait-related abnormalities in the concentration of monoaminergic synaptic terminals may be present in patients with asymptomatic bipolar disorder type I. METHOD: The concentration of a stable presynaptic marker, the vesicular monoamine transporter protein (VMAT2), was quantified with (+)[(11)C]dihydrotetrabenazine (DTBZ) and positron emission tomography. Sixteen asymptomatic patients with bipolar I disorder who had a prior history of mania with psychosis (nine men and seven women) and individually matched healthy subjects were studied. Correlational analyses were conducted to examine the relationship between regional VMAT2 binding, cognitive function, and clinical variables. RESULTS: VMAT2 binding in the thalamus and ventral brainstem of the bipolar patients was higher than that in the comparison subjects. VMAT2 concentrations in these regions correlated with performance on measures of frontal, executive function. In addition, sex differences in VMAT2 binding were detected in the thalamus of the bipolar patients; the male patients had higher binding than the women. No sex differences in binding were observed in the healthy comparison group. CONCLUSIONS: These initial results suggest that higher than normal VMAT2 expression and, by extension, concentration of monoaminergic synaptic terminals, may represent a trait-related abnormality in patients with bipolar I disorder and that male and female patients show different patterns. Also, VMAT2 concentrations may be associated with some of the cognitive deficits encountered in euthymic bipolar disorder. PMID- 11007717 TI - Factor analysis of the DSM-III-R borderline personality disorder criteria in psychiatric inpatients. AB - OBJECTIVE: The goal of this study was to examine the factor structure of the DSM III-R criteria for borderline personality disorder in young adult psychiatric inpatients. METHOD: The authors assessed 141 acutely ill inpatients with the Personality Disorder Examination, a semistructured diagnostic interview for DSM III-R personality disorders. They used correlational analyses to examine the associations among the different criteria for borderline personality disorder and performed an exploratory factor analysis. RESULTS: Cronbach's coefficient alpha for the borderline personality disorder criteria was 0.69. A principal components factor analysis with a varimax rotation accounted for 57.2% of the variance and revealed three homogeneous factors. These factors were disturbed relatedness (unstable relationships, identity disturbance, and chronic emptiness); behavioral dysregulation (impulsivity and suicidal/self-mutilative behavior); and affective dysregulation (affective instability, inappropriate anger, and efforts to avoid abandonment). CONCLUSIONS: Exploratory factor analysis revealed three homogeneous components of borderline personality disorder that may represent personality, behavioral, and affective features central to the disorder. Recognition of these components may inform treatment plans. PMID- 11007719 TI - Selective relationship between prefrontal N-acetylaspartate measures and negative symptoms in schizophrenia. AB - OBJECTIVE: Certain cognitive, behavioral, and emotional deficits (so-called negative symptoms) in patients with schizophrenia have often been attributed to prefrontal cortical pathology, but direct evidence for a relationship between prefrontal neuronal pathology and negative symptoms has been lacking. The authors hypothesized that an in vivo measure of prefrontal neuronal pathology (N: acetylaspartate [NAA] levels) in patients with schizophrenia would predict negative symptoms. METHOD: Proton magnetic resonance spectroscopic imaging ((1)H MRSI) and rating scales for negative and positive symptoms were used to study 36 patients with schizophrenia. Magnetic resonance spectra were analyzed as metabolite ratios, and parametric correlations were performed. RESULTS: A regionally selective negative correlation was found between prefrontal NAA creatine ratio and negative symptom ratings in this group of patients with schizophrenia. CONCLUSIONS: Lower prefrontal NAA-and by inference greater neuronal pathology-predicted more severe negative symptoms in patients with schizophrenia. These data demonstrate a relationship between an intraneuronal measure of dorsolateral prefrontal cortex integrity and negative symptoms in vivo and represent further evidence for the involvement of the dorsolateral prefrontal cortex in negative symptoms associated with schizophrenia. PMID- 11007718 TI - Brain activation patterns in schizophrenic and comparison volunteers during a matched-performance auditory recognition task. AB - OBJECTIVE: The biological characteristics of schizophrenia are often studied by using functional imaging techniques. However, since volunteers with schizophrenia routinely fail to perform as accurately or as quickly as healthy volunteers, it is difficult to ascertain whether a particular deficit in blood flow to a brain region is due to behavior or to the underlying illness. In this report, investigators used an auditory recognition task to assess brain blood flow patterns and behavioral correlates of schizophrenic patient volunteers trained on the task. METHOD: Twelve healthy volunteers and 18 volunteers with schizophrenia were trained to make tone frequency recognitions. Accuracy and stimuli were matched between groups. Participants were required to press a button to indicate whether a briefly presented tone was the high-frequency (1500 Hz) reference tone or one of a lower frequency level (level chosen to elicit an 80% accuracy score). Subjects underwent bolus [(15)O]H(2)O blood flow positron emission tomography during inactive rest, a sensory motor control condition, and the decision task. Blood flow patterns were assessed between conditions and between groups. RESULTS: As a group, the patients with schizophrenia (who performed as quickly and accurately as the comparison subjects) exhibited significantly less change in regional cerebral blood flow (rCBF) to the anterior cingulate and supplementary motor cortices when switching from the sensory motor control to the decision condition. There were also marked between-group differences in correlations between rCBF and response time. Whereas the comparison subjects exhibited progressively greater blood flow to the frontal cortex in association with longer response times, the schizophrenic patients exhibited progressively lower blood flow in conjunction with extended response times. CONCLUSIONS: The failure to appropriately enhance cingulate activity when engaged in a demanding task and the progressive, time-dependent decline in frontal blood flow suggest that patients with schizophrenia are unable to make optimal use of frontocingulate systems when maximally engaged in high-error tasks. PMID- 11007720 TI - Adult outcomes of child- and adolescent-onset schizophrenia: diagnostic stability and predictive validity. AB - OBJECTIVE: The goal of the study was to establish the predictive validity of a diagnosis of schizophrenia in childhood and early adolescence by examining diagnostic continuity into adult life and comparing social and symptomatic outcomes of child- and adolescent-onset schizophrenia with those of nonschizophrenic psychoses. METHOD: A total of 110 consecutive patients with first-episode child- or adolescent-onset psychosis (mean age at onset=14.2 years) presenting to the Maudsley Hospital in London between 1973 and 1991 were followed up an average of 11.5 years after first contact. Ninety-three (84.5%) of 110 patients were successfully followed-up, 51 with a first-episode diagnosis of DSM III-R schizophrenia and 42 with nonschizophrenic psychoses. Consensus best estimate DSM-III-R diagnoses were made at follow-up, and course and outcome were assessed blind to first-episode diagnosis. RESULTS: Diagnostic stability was high for child- and adolescent-onset DSM-III-R schizophrenia (positive predictive value=80%) and affective psychoses (positive predictive value=83%) but much lower for schizoaffective and atypical psychoses. Compared with other psychoses, child- or adolescent-onset schizophrenia was associated with significantly worse symptomatic and social outcomes, which were characterized by a chronic illness course and severe impairments in social relationships and independent living. CONCLUSIONS: The diagnosis of DSM-III-R schizophrenia in childhood and adolescence has good predictive validity. The high level of diagnostic stability suggests etiological continuity with adult schizophrenia, with onset in childhood and adolescence associated with a particularly malignant course and outcome. PMID- 11007721 TI - Modulation of the startle response and startle laterality in relatives of schizophrenic patients and in subjects with schizotypal personality disorder: evidence of inhibitory deficits. AB - OBJECTIVE: Patients with schizophrenia spectrum disorders have been shown to have deficits in sensorimotor gating as assessed by prepulse inhibition of the startle response. The authors hypothesized that nonschizophrenic relatives of patients with schizophrenia would also have prepulse inhibition deficits, thereby reflecting a genetically transmitted susceptibility to sensorimotor gating deficits. METHOD: Twenty-five comparison subjects, 23 patients with schizophrenia, 34 relatives of the schizophrenic patients, and 11 subjects with schizotypal personality disorder were assessed in an acoustic startle paradigm. The eye-blink component of the startle response was assessed bilaterally by using electromyographic recordings of orbicularis oculi. RESULTS: The patients with schizophrenia, their relatives, and subjects with schizotypal personality disorder all had reduced prepulse inhibition relative to comparison subjects, and these deficits were more evident in measures of right eye-blink prepulse inhibition. Comparison subjects demonstrated greater right versus left eye-blink prepulse inhibition, whereas the probands, their relatives, and subjects with schizotypal personality disorder showed less asymmetry of prepulse inhibition. CONCLUSIONS: These data suggest a genetically transmitted deficit in prepulse inhibition (sensorimotor gating) in patients with schizophrenia spectrum disorders, including subjects with schizotypal personality disorder and relatives of patients with schizophrenia. PMID- 11007722 TI - Relationship between climate and psychiatric inpatient length of stay in Veterans Health Administration hospitals. AB - OBJECTIVE: The study examined whether climate has an impact on inpatient psychiatric length of stay in Veterans Health Administration hospitals (VHA). METHOD: Data from the National Weather Service for eight climate variables for the locations of 134 VHA hospitals nationwide were factor analyzed, resulting in two climate factors representing temperature and precipitation. Factor scores were correlated with psychiatric mean lengths of stay from 1994 to 1998 for 99 VHA hospitals with inpatient psychiatric services and for the 22 VHA regional divisions (Veterans Integrated Service Networks). RESULTS: Climate factors correlated modestly but significantly with length of stay, with correlations ranging from -0. 25 to -0.37 at the hospital level and from -0.38 to -0.45 at the VHA regional level; hospitals in warmer and drier climates had shorter lengths of stay. Medical centers in colder climates had the longest lengths of stay in winter and fall. The significant correlation between climate and length of stay was not affected by recent reductions in length of stay in VHA hospitals. CONCLUSIONS: Higher clinical costs associated with longer lengths of stay in colder climates have implications for budget planning. Climate factors must also be recognized for their potential effect on performance monitoring systems focused on hospital utilization. Researchers must continue to consider broader contextual variables such as climate if they are to fully understand the determinants of health care utilization and psychiatric hospitalization costs. PMID- 11007723 TI - Seasonal variation of violence in Norway. AB - OBJECTIVE: Seasonal variations of violence have been the subject of some controversy. Norway, situated between latitudes 58 degrees and 72 degrees N, has considerable seasonal variations of light and provides a good opportunity for studies of seasonality. METHOD: The monthly numbers of police reports of violent incidents in 1991-1997 were obtained for the entire Norwegian population of 4,450,000 inhabitants and separately for each of seven Norwegian cities at different latitudes. RESULTS: A total of 82,537 episodes of violence were recorded. There was a significant variation in violent incidents between months, with a minimum daily frequency of 28.7 in March and a maximum daily frequency of 35.1 in June. The frequency curve had one significant peak in May through June and another significant peak in October through November. The monthly frequency of violence correlated with the absolute value of monthly change in length of day from the previous month. In the seven cities the highest monthly ratio of observed to expected frequencies increased with latitude. With increasing latitude, the months with the largest increase in violence came later both in the spring and in the fall. CONCLUSIONS: There is a distinct pattern of seasonal variation in the frequency of violence that varies systematically with latitude. This pattern resembles the seasonal pattern of some forms of suicide, hospitalization for affective disorders, and mood and activity in the general population. PMID- 11007724 TI - Associations between bipolar disorder and other psychiatric disorders during adolescence and early adulthood: a community-based longitudinal investigation. AB - OBJECTIVE: The study investigated cross-sectional and longitudinal associations between bipolar disorder and other psychiatric disorders during adolescence and early adulthood. METHOD: Psychiatric interviews were administered to a representative community sample of 717 youths and their mothers in 1983 (mean age of youths=14 years) and again in 1985-1986, and 1991-1993. RESULTS: A wide range of psychiatric disorders co-occurred with bipolar disorder during adolescence and early adulthood. Adolescent anxiety disorders were uniquely associated with increased risk for early adulthood bipolar disorder after adolescent bipolar disorder was accounted for. Manic symptoms during adolescence were associated with increased risk for anxiety and depressive disorders during early adulthood after adolescent anxiety and depressive disorders were accounted for. CONCLUSIONS: Adolescents with anxiety disorders may be at increased risk for bipolar disorder or clinically significant manic symptoms during early adulthood. Adolescents with manic symptoms may be at increased risk for anxiety and depressive disorders during early adulthood. PMID- 11007725 TI - Elevation in late-life depression of intercellular adhesion molecule-1 expression in the dorsolateral prefrontal cortex. AB - OBJECTIVE: Late-life depression may be associated with vascular disease. The authors investigated this association by determining whether intercellular adhesion molecule-1 (ICAM-1), a marker of ischemia-induced inflammation, is elevated in the dorsolateral prefrontal cortex in depression. METHOD: The authors studied postmortem tissue from 20 depressed subjects and a matched comparison group of 20 nondepressed subjects. They used immunocytochemistry to stain ICAM-1 in blood vessels on sections of the dorsolateral prefrontal cortex and occipital cortex and quantitative true color image analysis to measure the proportion of vessels expressing ICAM-1. RESULTS: ICAM-1 was significantly higher in both the gray and white matter of the depressed subjects' dorsolateral prefrontal cortex than the comparison subjects' dorsolateral prefrontal cortex. The difference between these groups was much smaller in the gray and white matter of the occipital cortex. CONCLUSIONS: These findings support the vascular depression hypothesis, which has important implications for the understanding and management of late-life depression. PMID- 11007726 TI - Use of no-suicide contracts by psychiatrists in Minnesota. AB - OBJECTIVE: The no-suicide contract is widely recommended as an important intervention in the care of suicidal patients; however, there are no data demonstrating its effectiveness or its acceptance in the professional community. This study examines the use of no-suicide contracts by psychiatrists in Minnesota. METHOD: A postcard questionnaire was mailed to 514 psychiatrists in Minnesota inquiring about their practices and experiences with no-suicide contracts. RESULTS: There were 267 responses, yielding a response rate of 52%. No suicide contracts were used by 152 (57%) of the respondents. Within this group, 62 (41%) of the psychiatrists had patients who committed suicide or made serious attempts after entering into a no-suicide contract. CONCLUSIONS: Among the respondents to the questionnaire, slightly more than half used no-suicide contracts, indicating that such contracts are not universally accepted as standard practice among these psychiatrists. More data are needed to determine the effectiveness of no-suicide contracts in preventing suicide. PMID- 11007727 TI - Randomized, placebo-controlled, double-blind clinical trial of sertraline in the treatment of depression complicating Alzheimer's disease: initial results from the Depression in Alzheimer's Disease study. AB - OBJECTIVE: This study evaluated the efficacy and safety of sertraline in the treatment of major depression in 22 outpatients with Alzheimer's disease. METHOD: Twelve of the 22 patients were given sertraline and 10 were given placebo by random group assignment for 12 weeks. Response to treatment was measured by using the Cornell Scale for Depression in Dementia. The patients were also assessed with the Hamilton Depression Rating Scale, the activities of daily living subscale of the Psychogeriatric Dependency Rating Scales, and the Mini-Mental State. RESULTS: After 12 weeks of double-blind, placebo-controlled treatment, nine of the patients given sertraline and two of those given placebo were at least partial responders. Patients given sertraline had significantly greater mean declines from baseline in Cornell Scale for Depression in Dementia scores; the bulk of antidepressant response occurred by the third week of treatment. CONCLUSIONS: Sertraline is superior to placebo in reducing depression in patients with Alzheimer's disease who also suffer from major depression. PMID- 11007728 TI - Thyroid hormone levels and recurrence of major depression. AB - OBJECTIVE: The relationship between basal thyroid hormone levels and acute antidepressant response has been studied, but any relationship between basal thyroid hormone levels and long-term course of depressive illness has not been evaluated. METHOD: The authors used a Cox regression survival analysis to evaluate the relationship between life course of depressive illness and basal levels of thyroid hormones (triiodothyronine [T(3)], thyroxine [T(4)], and thyrotropin) in 75 outpatients with unipolar major depressive disorder. RESULTS: Time to recurrence of major depression was inversely related to T(3) levels but not to T(4) levels. CONCLUSIONS: These data may be of clinical interest in view of the fact that T(3) is used to augment antidepressant response. PMID- 11007729 TI - Functional anatomy of auditory verbal imagery in schizophrenic patients with auditory hallucinations. AB - OBJECTIVE: This study investigated the functional neuroanatomy of inner speech and auditory verbal imagery in schizophrenic patients predisposed to auditory hallucinations. METHOD: Eight patients with schizophrenia with a history of prominent auditory hallucinations and six comparison subjects underwent functional magnetic resonance imaging while generating inner speech or imagining external speech. RESULTS: Patients showed no differences while generating inner speech but experienced a relatively attenuated response in the posterior cerebellar cortex, hippocampi, and lenticular nuclei bilaterally and the right thalamus, middle and superior temporal cortex, and left nucleus accumbens during auditory verbal imagery. CONCLUSIONS: Patients with schizophrenia who were prone to auditory hallucinations show attenuated activation when processing inner speech in areas implicated in verbal self-monitoring. PMID- 11007730 TI - Preliminary evidence of improved verbal working memory performance and normalization of task-related frontal lobe activation in schizophrenia following cognitive exercises. AB - OBJECTIVE: The authors' goal was to evaluate the possibility of treating brain and behavioral aspects of verbal memory dysfunction in patients with schizophrenia through cognitive exercises. METHOD: Eight patients did daily verbal memory exercises that became progressively more difficult over a 10-week training period. Memory performance and regional brain activations during a verbal memory task were assessed before and after these exercises. RESULTS: Verbal but not nonverbal memory performance improved after training; three patients made substantial gains, and five showed little change. Performance gains were correlated with increases in task-related activation of the left inferior frontal cortex. One patient given 5 extra weeks of training 6 weeks after the initial training period showed maintenance of initial performance gains 6 weeks after training, further improvement after the second period of training, and normalization of task-related activation of the left inferior frontal cortex. CONCLUSIONS: Verbal memory deficits can be ameliorated by memory exercises in some patients with schizophrenia. Performance improvements are associated with increased task-related activation of the same brain region that is activated during verbal memory tasks in healthy individuals. PMID- 11007731 TI - Effects of methylphenidate on functional MRI blood-oxygen-level-dependent contrast. AB - OBJECTIVE: The authors' goal was to determine potential hemodynamic consequences of methylphenidate on functional magnetic resonance imaging (MRI) blood-oxygen level-dependent (BOLD) contrast. METHOD: BOLD and perfusion changes were recorded from the motor cortex of six healthy subjects while they performed flexion extension movements of the right index finger (finger tapping) at varying rates before and after oral methylphenidate administration. RESULTS: Functional MRI signals increased monotonically with faster movement rates. Subjects' heart rates increased modestly after methylphenidate administration, but no changes in finger tapping performance or functional MRI signals were observed. CONCLUSIONS: Methylphenidate does not alter BOLD neural-hemodynamic coupling. Consequently, functional MRI can be used to map neural systems that subserve cognitive operations (e.g., attention and executive processes) in subjects taking methylphenidate. PMID- 11007733 TI - High doses of levothyroxine for refractory rapid cycling. PMID- 11007732 TI - Prediction of dopamine transporter binding availability by genotype: a preliminary report. AB - OBJECTIVE: Evidence of a relationship between genotype and binding availability was assessed for the dopamine and serotonin transporter genes. METHOD: The authors assessed dopamine transporter genotype at the SLC6A3 3' variable number of tandem repeats (VNTR) polymorphism and serotonin transporter genotype at the SLC6A4 promotor VNTR polymorphism in 30 healthy subjects who also underwent single photon emission computed tomography with [(123)I]beta-CIT. RESULTS: Subjects homozygous for the 10-repeat allele at the SLC6A3 locus demonstrated significantly lower dopamine transporter binding than carriers of the nine-repeat allele. There was no effect of SLC6A4 genotype upon serotonin transporter binding. CONCLUSIONS: These findings suggest that genetic variation at the SLC6A3 3' VNTR polymorphism may modify dopamine transporter function. PMID- 11007735 TI - Agranulocytosis associated with lamotrigine. PMID- 11007736 TI - Delirium with autoimmune thyroiditis induced by interferon alpha. PMID- 11007737 TI - Citalopram and haloperidol for psychotic depression. PMID- 11007738 TI - gamma-Hydroxybutyrate withdrawal and chloral hydrate. PMID- 11007739 TI - Adenosine and antidepressant effects of sleep deprivation. PMID- 11007740 TI - Psychological defense styles and childhood sexual abuse. PMID- 11007741 TI - Measuring dopamine D(2) receptors. PMID- 11007744 TI - Serotonin 2A receptor polymorphisms and [(3)H]Ketanserin binding. PMID- 11007745 TI - Treatment of patients with delirium. PMID- 11007754 TI - Immunological transitions in response to antigenic mutation during viral infection. AB - Antigenic variation is an important factor in viral persistence and disease progression. We analyze immunological changes which occur in response to antigenic mutation during chronic viral infection. Using an established model of viral and immune system dynamics, we determine which qualitative shifts in the immune response can be elicited by the appearance of a new mutant. We find that antigenic mutation can cause dramatic shifts in the magnitude and type of anti viral immune response. For example, the appearance of a mutant can elicit a new immune response which recognizes the original viral strain. We also find that novel strains of the virus which replicate more slowly than existing viral strains are able to invade and survive, even when the immune system is capable of mounting an immune response against the mutant. PMID- 11007755 TI - Tumor necrosis factor receptor-1 is critically involved in the development of experimental autoimmune myasthenia gravis. AB - Tumor necrosis factor receptor-1 (TNFR1, CD120a) has been implicated in the pathogenesis of several experimental models of T cell-mediated autoimmune disorders, but its role in antibody-mediated autoimmune diseases has not been addressed. Experimental autoimmune myasthenia gravis (EAMG), an autoantibody mediated T cell-dependent neuromuscular disorder, represents an animal model for myasthenia gravis in human. To investigate the role of TNFR1 in the pathogenesis of EAMG, TNFR1(-/-) and wild-type mice were immunized with TORPEDO: acetylcholine receptor (AChR) in complete Freund's adjuvant. TNFR1(-/-) mice failed to develop EAMG. Lymphoid cells from TNFR1(-/-) mice produced low amounts of T(h)1 (IFN gamma, IL-2 and IL-12)-type cytokines, but elevated levels of T(h)2 (IL-4 and IL 10)-type cytokines compared with lymphoid cells of wild-type mice. Accordingly, the levels of anti-AChR IgG2 antibodies were severely reduced and the level of anti-AChR IgG1 antibodies were moderately reduced. Co-injection of recombinant mouse IL-12 with AChR in adjuvant restored T cell responses to AChR and promoted development of EAMG in TNFR1(-/-) mice. These results demonstrate that the TNF/TNFR1 system is required for the development of EAMG. The lack of a functional TNF/TNFR1 system can, at least in part, be substituted by IL-12 at the stage of initial priming with AChR and adjuvant. PMID- 11007756 TI - The Pim-1 kinase stimulates maturation of TCRbeta-deficient T cell progenitors: implications for the mechanism of Pim-1 action. AB - We demonstrate that overexpression of Pim-1, a cytoplasmic serine/threonine kinase of poorly defined function, results in the development of substantial numbers of CD4(+)CD8(+) double-positive thymocytes in two independent knock-out mouse models (i.e. the RAG-1-deficient and TCRbeta gene enhancer-deleted mice) in which production of a functionally rearranged TCRbeta gene (hence the pre-TCR) is impaired. This activity of Pim-1, however, does not affect signaling through the Ras/Raf/MAP kinase cascade nor signaling which mediates suppression of TCRbeta gene recombination (i.e. allelic exclusion). While overexpression of Pim-1 positively affects cell cycle progression in selected CD4(-)CD8(-) double negative precursors, it did not affect expression of components of the cell cycle machinery, with the exception of the G(1)-specific phosphatase Cdc25A upon antigen receptor stimulation. We propose that Pim-1 acts downstream, or in parallel, to pre-TCR-mediated selection as one factor involved in the proliferative expansion of beta-selected pre-T cells. PMID- 11007757 TI - Identification and characterization of a molecule, BAM11, that associates with the pleckstrin homology domain of mouse Btk. AB - Bruton's tyrosine kinase (Btk) is required for normal B cell development and signal transduction through cell surface molecules, and its defects lead to X linked immune deficiency in mice and X-linked agammaglobulinemia in humans. In this report, we will describe the identification and characterization of a molecule, BAM11, which binds to the pleckstrin homology domain of Btk. A sequence homology search revealed that BAM11 has 89% homology, at the amino acid level, to human LTG19/ENL, that was originally identified as one of the fusion partners involved in chromosomal translocations of 11q23, MLL/ALL-1/HRX, in leukemia cells. Deletion mutants demonstrated that the region of BAM11 required for binding to Btk was localized between amino acid residues 240 and 256. Forced expression of a truncated form of BAM11 (amino acids 246-368) inhibited IL-5 induced proliferation by 50%, whereas forced expression of full-length BAM11 in Y16 cells did not affect the IL-5 responsiveness. We have also shown that BAM11 (amino acids 246-368) inhibited the kinase activity of Btk. These results suggest that the binding of BAM11 to Btk plays a regulatory role in the Btk signal transduction pathway. A cell fractionation study and analysis using EGFP-fused Btk protein demonstrated that a proportion of Btk is present within the nucleus. PMID- 11007758 TI - Blocked transport of soluble K(b) molecules containing connecting peptide segment involved in calnexin association. AB - The molecular event governing the assembly of the MHC class I heavy chain-beta(2) microglobulin-peptide complex is still not fully understood. In order to characterize the transport properties of MHC class I molecules, several truncated H-2K(b) genes were constructed and expressed in COS7 cells. Surprisingly, the expressed soluble molecule containing connecting peptide (CP) segment (sK(b)(CP)) did not secrete as efficiently as the one without CP (sK(b)(CYT)). When the sK(b)(CP) gene was transfected into a calnexin-deficient cell line CEM.NK(R), the amount of soluble K(b) molecules in the supernatant was comparable with sK(b)(CYT)-transfected CEM.NK(R). To further demonstrate the different transport of sK(b)(CP) and sK(b)(CYT) within living cells, we attached green fluorescent protein (GFP) to the C-termini of both molecules and, as a comparison, to the full-length transmembrane counterpart (mK(b)-GFP). While the mK(b)-GFP transfected cells showed the green fluorescence in the reticular network and the nuclear envelope, sK(b)(CP)-GFP showed obviously lump fluorescence of high intensity within cells. However, the distribution of sK(b)(CYT)-GFP was fairly uniform. Furthermore, GFP-tagged molecules allow us to analyze their interaction with other proteins in a direct, simple and quantitative method, designated immunofluorescence precipitation. The results showed that 60% of sK(b)(CP)-GFP molecules were associated with calnexin, while <10% with tapasin. Taken together with the results from sK(b)(CYT)-GFP and mK(b)-GFP, it is reasonable to deduce that the CP segment is involved in the association of class I molecules with calnexin and the transmembrane region might play a dynamic role in the dissociation from calnexin. The suggested kinetic association of class I molecules with calnexin is likely to contribute to the different maturation rate between several class I alleles. PMID- 11007759 TI - Negative regulation of B cell receptor-mediated signaling in B-1 cells through CD5 and Ly49 co-receptors via Lyn kinase activity. AB - CD5(+) B-1 cells are known to be unresponsive to B cell receptor (BCR)-mediated growth signals but instead undergo apoptosis. However, the B-1 cells from Lyn kinase-deficient (Lyn-/-) mice exhibited an enhanced proliferative response upon BCR cross-linking. It has been reported that BCR-mediated signaling in B-1 cells is negatively regulated by signals from CD22, CD5 and CD72 co-receptors, and that Lyn kinase plays a crucial role in tyrosine phosphorylation of immunoreceptor tyrosine-based inhibitory motifs on the CD22 and CD72, which recruits SHP-1 to the BCR complex. We found that Lyn kinase is also essential for the tyrosine phosphorylation of CD5 and subsequent recruitment of SHP-1 in B-1 cells upon cross-linking of BCR. Moreover, a distinct subpopulation of B-1 cells was found to express cell surface Ly49, which is known as a MHC class I-binding negative regulatory receptor on NK cells. Ly49 was rapidly tyrosine phosphorylated upon cross-linking of BCR and SHP-1 was found to recruit to the phosphorylated Ly49. Addition of F(ab')(2) fragments of anti-Ly49 antibodies partially blocked negative signals in B-1 cells. Thus two co-receptors, CD5 and Ly49, which are unique to B-1 cells, play a role in the regulation of B-1 cell activation. These results indicate that BCR-mediated signals in B-1 cells are strictly and negatively regulated through multiple pathways, that are dependent on Lyn kinase activity. PMID- 11007760 TI - Contribution of H-2 and non-H-2 genes in the control of mercury-induced autoimmunity. AB - Mercury-induced autoimmunity is characterized by a T cell-dependent B cell activation (mainly of IgG1 and IgE isotypes), production of anti-nucleolar autoantibodies (ANolA) and the formation of renal IgG deposits. The autoimmunity is to a large extent controlled by genetic factors. We studied 15 different inbred mouse strains of seven H-2 (mouse MHC) genotypes to determine the importance of H-2 and non-H-2 background genes in mercury-induced autoimmunity. The tested strains exhibited a diverse autoimmune response to mercury. In each H 2 genotype, there was at least one strain which responded to mercury by the production of high levels of IgG1 and IgE Ig as well as by the development of high titers of renal IgG1 deposits. Only mouse strains with H-2(s) and H-2(q) genotypes, irrespective of their background genes, produced ANolA after mercury treatment. Only SJL (H-2(s)) and A.SW (H-2(s)) mice were highly susceptible to all characteristics of mercury-induced autoimmunity. NZB (H-2(d)) mice were also highly susceptible, but they did not develop ANolA. Only the DBA/2 (H-2(d)) strain was found to be resistant to all tested mercury-induced autoimmune manifestations, suggesting that non-responsiveness to mercury in DBA/2 mice was largely influenced by non H-2 genes. These findings imply that H-2 genes mainly determine the susceptibility to mercury-induced ANolA production, whereas non-H-2 genes control the susceptibility to and the severity of the B cell activation and renal IgG deposition. PMID- 11007761 TI - Differences in specificities of anti-centromere sera for the monomeric and dimeric C-terminal peptides of human centoromere protein C. AB - Centromere protein-C (CENP-C), one of the centromere autoantigens and components of the inner plate of the kinetochore, is suggested to make a dimer at the C terminus. In order to investigate the presence of conformation-specific anti centromere antibodies (ACA) to the dimer form, the C-terminal 124 amino acids (CF 124) were expressed in Escherichia coli, affinity purified and chemically cross linked. Immunoblotting was utilized to compare the reactivities between the dimers and the monomers against 58 ACA(+) sera. The reactivities of the dimers were obviously higher in both IgG and IgM responses. The dimer was still more reactive than the glutathione S-transferase-fused monomer in some sera. Two kinds of CF-124 mutant (each contained one amino acid change at the N-terminal region of CF-124) and two cut segments of CF-124 (67 N-terminal amino acids and 58 C terminal amino acids) were also examined. The former two mutants decreased the dimerization activity. The latter two mutants lost both activities except for the faint dimerization activity of the N-terminal half. Affinity-purified antibodies with CF-124 in a liquid phase containing the co-purified GroE protein of E. coli, GroEL, reacted to the centromere in culture cells. In conclusion, there are heterogeneous autoepitopes including some conformational epitopes at the C terminal CENP-C. PMID- 11007762 TI - Characterization of a new human B7-related protein: B7RP-1 is the ligand to the co-stimulatory protein ICOS. AB - Optimal T cell activation requires the interactions of co-stimulatory molecules, such as those in the CD28 and B7 protein families. Recently, we described the co stimulatory properties of the murine ligand to ICOS, which we designated as B7RP 1. Here, we report the co-stimulation of human T cells through the human B7RP-1 and ICOS interaction. This ligand-receptor pair interacts with a K:(D) approximately 33 nM and an off-rate with a t((1/2)) > 10 min. Interestingly, tumor necrosis factor (TNF)-alpha differentially regulates the expression of human B7RP-1 on B cells, monocytes and dendritic cells (DC). TNF-alpha enhances B7RP-1 expression on B cells and monocytes, while it inhibits it on DC. The human B7RP-1-Fc protein or cells that express membrane-bound B7RP-1 co-stimulate T cell proliferation in vitro. Specific cytokines, such as IFN-gamma and IL-10, are induced by B7RP-1 co-stimulation. Although IL-2 levels are not significantly increased, B7RP-1 co-stimulation is dependent on IL-2. These experiments define the human ortholog to murine B7RP-1 and characterize its interaction with human ICOS. PMID- 11007763 TI - Oral administration of cholera toxin B subunit conjugated to myelin basic protein protects against experimental autoimmune encephalomyelitis by inducing transforming growth factor-beta-secreting cells and suppressing chemokine expression. AB - The efficacy and mechanism of immunosuppression against experimental autoimmune encephalomyelitis (EAE) by oral low-dose administration of myelin basic protein (MBP) conjugated to cholera toxin B subunit (CTB) were investigated in Lewis rats immunized with MBP together with complete Freund's adjuvant 4 days before the start of treatment. Oral treatment with CTB-MBP conjugate gave almost complete protection against disease, an effect that was totally abrogated by including a low dose of cholera holotoxin (CT). The protection by CTB-MBP was associated with a dramatic reduction in the number of leukocytes staining for CD4, CD8, IL-2R or MHC class II in the spinal cord as examined by immunohistochemistry. The mRNA expressions of T(h)1 cytokines IFN-gamma, IL-12 and tumor necrosis factor-alpha, as well as of chemokines monocyte chemotactic protein (MCP)-1 and RANTES in the spinal cord were also reduced by 76-94%, as assessed by in situ hybridization. In contrast, transforming growth factor (TGF)-beta mRNA-expressing cells were strongly increased in the spinal cord from animals treated orally with the CTB MBP conjugate. In the draining peripheral lymph nodes, the number of MBP-specific TGF-beta mRNA-expressing cells was also increased, whereas there was a decrease in cells expressing T(h)1 or T(h)2 cytokine mRNA. Protection against EAE could be transferred by injection of cells from the mesenteric lymph nodes of animals fed with CTB-MBP into naive animals exposed to encephalitogenic T cells. The results indicate that the protective anti-inflammatory effect by oral treatment with CTB MBP conjugate is, to a large extent, due to the induction of TGF-beta-secreting suppressive-regulatory T cells and to local down-regulation of MCP-1 and RANTES in the spinal cord. PMID- 11007764 TI - The level of expression of mu heavy chain modifies the composition of peripheral B cell subpopulations. AB - The B cell receptor (BCR) has a decisive role in transducing signals required for the development of B cells and their survival in the periphery. However, the processes that initiate these signals remain unclear and concepts of constitutive and ligand-dependent signaling have been proposed. Using a mu-transgenic mouse model, we have analyzed the impact of high surface IgM expression on the composition of the splenic B cell population. kappa-deficient mice homozygous for the H3-mu transgene have B cells with a higher BCR surface density than H3 heterozygous mice. This higher BCR expression is associated with an increase in the percentage and the total number of splenic B cells. In addition, an important proportion of CD23(-)CD21(+) marginal zone (MZ) B cells can be observed in H3 homozygous mice. However, these modifications operate in the absence of impairment of the positive selection process of the H3-mu/lambda1 combination over the H3-mu/lambda2 + 3 ones. These results suggest that (i) a constitutive BCR signaling directly correlated with BCR surface density is responsible for the efficient B cell colonization of the periphery with an accumulation of B cells in the MZ and (ii) a ligand-dependent BCR signal is responsible for the clonotype composition of the mature B cell repertoire. PMID- 11007765 TI - Neonatal induction of tolerance to T(h)2-mediated autoimmunity in rats. AB - Brown-Norway (BN) rats are highly susceptible to drug-induced immune dysregulations and when injected with mercuric chloride (HgCl(2)) or sodium aurothiopropanolsulfonate (ATPS), they develop a syndrome characterized by a polyclonal B cell activation depending upon CD4(+) T(h)2 cells that recognize self-MHC class II molecules. Since peripheral tolerance of T(h)2 cells might be crucial in the prevention of immunological manifestations such as allergy, establishing conditions for inducing tolerance to HgCl(2)- or ATPS-mediated immune manifestations appeared to be of large interest. We report here that BN rats neonatally injected with HgCl(2): (i) do not develop the mercury disease, (ii) remain resistant to HgCl(2)-induced autoimmunity at 8 weeks of age and later, provided they are regularly exposed to HgCl(2), (iii) are still susceptible to ATPS-induced immune manifestations, and (iv) exhibit spleen cells that adoptively transfer tolerance to HgCl(2)-induced autoimmunity in naive, slightly irradiated, syngeneic recipients. These findings demonstrate that dominant specific tolerance can be neonatally induced using a chemical otherwise responsible for T(h)2-mediated autoimmunity. PMID- 11007766 TI - In vivo cytotoxicity of the prion protein fragment 106-126. AB - Transmissible spongiform encephalopathies are fatal neurological diseases characterized by astroglyosis, neuronal loss, and by the accumulation of the abnormal isoform of the prion protein. The amyloid prion protein fragment 106-126 (P106-126) has been shown to be toxic in cultured hippocampal neurons (). Here, we show that P106-126 is also cytotoxic in vivo. Taking advantage of the fact that retina is an integral part of the central nervous system, the toxic effect of the peptide was investigated by direct intravitreous injection. Aged solutions of P106-126 induced apoptotic-mediated retinal cell death and irreversibly altered the electrical activity of the retina. Neither apoptosis nor electroretinogram damages were observed with freshly diluted P106-126, suggesting that the toxicity is linked to the aggregation state of the peptide. The retina provides a convenient in vivo system to look for potential inhibitors of cytotoxicity associated with spongiform encephalopathies. PMID- 11007767 TI - A positive role for the PP2A catalytic subunit in Wnt signal transduction. AB - Protein phosphatase-2A (PP2A) is a multisubunit serine/threonine phosphatase involved in intracellular signaling, gene regulation, and cell cycle progression. Different subunits of PP2A bind to Axin and Adenomatous Polyposis Coli, components of the Wnt signal transduction pathway. Using early Xenopus embryos, we studied how PP2A functions in Wnt signal transduction. The catalytic subunit of PP2A (PP2A-C) potentiated secondary axis induction and Siamois reporter gene activation by Dishevelled, a component of the Wnt pathway, indicating a positive regulatory role of this enzyme in Wnt signaling. In contrast, small t antigen, an antagonist of PP2A-C, inhibited Dishevelled-mediated signal transduction, as did the regulatory PP2A-B'epsilon subunit, consistent with the requirement of PP2A function in this pathway. Although Wnt signaling is thought to occur via regulation of beta-catenin degradation, PP2A-C did not significantly affect beta catenin stability. Moreover, the pathway activated by a stabilized form of beta catenin was sensitive to PP2A-C and its inhibitors, suggesting that PP2A-C acts downstream of beta-catenin. Because previous work has suggested that PP2A can act upstream of beta-catenin, we propose that PP2A regulates the Wnt pathway at multiple levels. PMID- 11007768 TI - Reversal of autocrine and paracrine effects of interleukin 1 (IL-1) in human arthritis by type II IL-1 decoy receptor. Potential for pharmacological intervention. AB - Interleukin 1 (IL-1), produced by both synovial cells and chondrocytes, plays a pivotal role in the pathogenesis of cartilage destruction in osteoarthritis (OA). We examined the specific expression and function of IL-1 receptor family-related genes in human joint tissues. Gene array analysis of human normal and OA-affected cartilage showed mRNA expression of IL-1 receptor accessory protein (IL-1RAcp) and IL-1 type I receptor (IL-1RI), but not IL-1 antagonist (IL-1ra) and IL-1 type II decoy receptor (IL-1RII). Similarly, human synovial and epithelial cells showed an absence of IL-1RII mRNA. Functional genomic analyses showed that soluble (s) IL-1RII, at picomolar concentrations, but not soluble TNF receptor:Fc, significantly inhibited IL-1beta-induced nitric oxide (NO) and/or prostaglandin E(2) production in chondrocytes, synovial and epithelial cells. In OA-affected cartilage, the IC(50) for inhibition of NO production by sIL-1RII was 2 log orders lower than that for sIL-1RI. Human chondrocytes that overexpressed IL-1RII were resistant to IL-1-induced IL-1beta mRNA accumulation and inhibition of proteoglycan synthesis. In osteoarthritis, deficient expression by chondrocytes of innate regulators or antagonists of IL-1 such as IL-1ra and IL 1RII (soluble or membrane form) may allow the catabolic effects of IL-1 to proceed unopposed. The sensitivity of IL-1 action to inhibition by sIL-1RII has therapeutic implications that could be directed toward correcting this unfavorable tissue(s) dependent imbalance. PMID- 11007769 TI - Goodpasture antigen-binding protein, the kinase that phosphorylates the goodpasture antigen, is an alternatively spliced variant implicated in autoimmune pathogenesis. AB - The non-collagenous C-terminal domain of the alpha(3) chain of collagen IV is the autoantigen in Goodpasture disease, an autoimmune disorder described only in humans. Specific N-terminal phosphorylation is a biological feature unique to the human domain when compared with other homologous domains lacking immunopathogenic potential. We have recently cloned from a HeLa-derived cDNA library a novel serine/threonine kinase (Goodpasture antigen-binding protein (GPBP)) that phosphorylates the N-terminal region of the human domain (Raya, A. Revert, F, Navarro, S. and Saus J. (1999) J. Biol. Chem. 274, 12642-12649). We show here that the pre-mRNA of GPBP is alternatively spliced in human tissues and that the most common transcript found encodes GPBPDelta26, a molecular isoform devoid of a 26-residue serine-rich motif. Recombinantly expressed GPBPDelta26 exhibits lower activity than GPBP, due at least in part to a reduced ability of GPBPDelta26 to interact and to form very active high molecular weight aggregates. In human tissues, GPBP shows a more limited expression than GPBPDelta26 but displays a remarkable preference for the small vessels and for histological structures targeted by natural autoimmune responses including alveolar and glomerular basement membranes, the two main targets in Goodpasture disease. GPBP expression is, in turn, up-regulated in the striated muscle of a Goodpasture patient and in other autoimmune conditions including cutaneous lupus erythematosus, pemphigoid, and lichen planus. PMID- 11007770 TI - Synergistic cooperation between Sp1 and Smad3/Smad4 mediates transforming growth factor beta1 stimulation of alpha 2(I)-collagen (COL1A2) transcription. AB - Transforming growth factor-beta1 (TGFbeta) is a strong activator of extracellular matrix accumulation. TGFbeta stimulates the gene coding for human alpha2(I) collagen (COL1A2) by inducing binding of an Sp1-containing complex to an upstream promoter element (TGFbeta responsive element or TbRE) that contains a CAGA box. Here we report that the CAGA box of the TbRE is the binding site of the Smad3/Smad4 complex, and that the binding of the complex is required for TGFbeta induced COL1A2 up-regulation. Recombinant Smad3 and Smad4 bind in vitro to the CAGA box of COL1A2; TGFbeta treatment of cultured fibroblasts induces Smad3/Smad4 binding to the TbRE; transient overexpression of Smad3 and Smad4 in fibroblasts transactivates TbRE-driven transcription; and COL1A2 gene up-regulation by TGFbeta is abolished in cells stably transfected with plasmids that express dominant negative forms of Smad3 or Smad4. In Sp1-deficient Drosophila Schneider cells, there was cooperative synergy between Smad3/Smad4 and Sp1 at the TbRE site. The analysis also emphasized the requirement of both Sp1- and Smad-binding sites for optimal promoter transactivation. In cells stably transfected with a plasmid expressing a dominant negative form of Sp1, the synergy was shown to be promoter-specific and dependent on the binding of Sp1 to the TbRE. Interestingly, overexpression of dominant negative Sp1 was found to block the antagonistic signal of tumor necrosis factor-alpha on COL1A2 transcription, as well. These results provide the first linkage between the Smad3 and Smad4 proteins and TGFbeta stimulation of type I collagen biosynthesis. PMID- 11007771 TI - Cell adhesion regulates ubiquitin-mediated degradation of the platelet-derived growth factor receptor beta. AB - Cross-talk between integrin-mediated adhesion and growth factors has been described in many recent studies; however, the underlying mechanisms remain incompletely understood. We report here that detachment of cells from the extracellular matrix induced a decrease in both the autophosphorylation and protein levels of the platelet-derived growth factor receptor beta (PDGF-R beta), which was completely reversed upon replating cells on fibronectin. The effect occurred in all cells examined but to a greater extent in primary fibroblasts compared with established cell lines. Decreased PDGF-R levels in suspended cells correlated with ubiquitination of the PDGF-R and was blocked by treatment with inhibitors of the proteasome pathway. Unlike PDGF-induced down-regulation, detachment-induced degradation did not require receptor autophosphorylation, internalization, or tyrosine kinase activity. We conclude that cell detachment results in cellular desensitization to PDGF that is mediated by degradation of the PDGF-R via a novel ubiquitin-dependent pathway. PMID- 11007772 TI - Expression of constitutively active phosphatidylinositol 3-kinase inhibits activation of caspase 3 and apoptosis of cardiac muscle cells. AB - Apoptosis of cardiac muscle cells contributes to the development of cardiomyopathy. Recent studies showed that insulin-like growth factor I (IGF-I) inhibits apoptosis of cardiac muscle cells and improves myocardial function in experimental heart failure. This study was carried out to elucidate the role of phosphatidylinositol 3-kinase (PI 3-kinase) in the anti-apoptotic actions of IGF I in cardiomyocytes and to explore whether expression of constitutively active PI 3-kinase can inhibit apoptosis in cardiomyocytes. Apoptosis of primary cardiomyocytes was induced by doxorubicin treatment and serum withdrawal. Transduction of cardiomyocytes with constitutively active PI 3-kinase specifically lead to serine phosphorylation of Akt, whereas phosphorylation of IGF-I receptor, IRS1/2 and p44/42 mitogen-activated protein kinase were not increased. In the cardiomyocytes transduced with constitutively active PI 3 kinase, activation of the pro-apoptotic caspase 3 was attenuated and fragmentation of DNA was reduced. Preincubating cells with PI 3-kinase inhibitor LY294002 was associated with loss of anti-apoptotic actions of IGF-I and PI 3 kinase. Neither IGF-I nor constitutively active PI 3-kinase lead to serine phosphorylation of Bad, suggesting that the anti-apoptotic effects of PI 3-kinase are not mediated through Bad phosphorylation in cardiac muscle cells. To determine whether activation of caspase 3 is sufficient to induce apoptosis in cardiomyocytes, an engineered TAT-caspase 3 protein was introduced to cardiomyocytes. Significant reduction of cell viability occurred in the cardiomyocytes transduced with active caspase 3, indicating that activation of caspase 3 is sufficient to cause cardiomyocyte death. These findings indicate the existence of an IGF-I receptor-PI 3-kinase-caspase 3 pathway in cardiomyocytes that plays an important role in the anti-apoptotic actions of IGF-I in heart. Moreover, these data suggest that modulation of PI 3-kinase activities may represent a potential therapeutic strategy to counteract the occurrence of apoptosis in cardiomyopathy. PMID- 11007773 TI - Transcriptional activities of reovirus RNA polymerase in recoated cores. Initiation and elongation are regulated by separate mechanisms. AB - The particle-associated reovirus polymerase synthesizes mRNA within only certain viral particle types. Reovirus cores, subviral particles lacking outer capsid proteins mu1, sigma3, and sigma1, produce mRNA and abortive transcripts. Reovirus virions, which contain complete outer capsids, cannot produce mRNA and produce few abortive transcripts. Recoated cores are virion-like particles generated by the addition of recombinant outer capsid proteins to cores. We used recoated cores to analyze transcriptional regulation by reovirus outer capsid proteins. Partially recoated particles, containing less than virion amounts of mu1 and sigma3, synthesized mRNA at levels inversely proportional to outer capsid protein levels. Fully recoated cores exhibited undetectable mRNA synthesis levels, as did virions. However, recoated cores produced high levels of abortive transcripts. Recoated core abortive transcripts remained particle-associated and appeared to inhibit further abortive transcript production. Proteolysis of recoated cores removing mu1 and sigma3 released accumulated abortive transcripts and relieved inhibition of mRNA and abortive transcript synthesis. These results suggest transcriptional elongation, but not initiation, is blocked by virion-like amounts of mu1 and sigma3. Particle-associated abortive transcripts may down-regulate transcriptional initiation. Minor outer capsid protein sigma1 had no demonstrable effect on transcriptional activities. Transcriptional regulation may ensure progeny virions do not compete with transcribing particles for ribonucleoside triphosphates. PMID- 11007774 TI - Identification of protein-tyrosine phosphatase 1B as the major tyrosine phosphatase activity capable of dephosphorylating and activating c-Src in several human breast cancer cell lines. AB - c-Src tyrosine kinase activity is elevated in several types of human cancer, and this has been attributed to elevated c-Src expression levels, increased c-Src specific activity, and activating mutations in c-Src. We have found a number of human breast cancer cell lines with elevated c-Src specific activity that also possess elevated phosphatase activity directed against the carboxyl-terminal negative regulatory domain of Src family kinases. To identify this phosphatase, cell extracts from MDA-MB-435S cells were chromatographed and the fractions were assayed for phosphatase activity. Four peaks of phosphatase activity directed against the nonspecific substrate poly(Glu/Tyr) were detected. One peak also dephosphorylated a peptide modeled against the c-Src carboxyl-terminal negative regulatory domain and intact human c-Src. Immunoblotting and immunodepletion experiments identified the phosphatase as protein-tyrosine phosphatase 1B (PTP1B). Examination of several human breast cancer cell lines with increased c Src activity showed elevated levels of PTP1B protein relative to normal control breast cells. In vitro c-Src reactivation experiments confirmed the ability of PTP1B to dephosphorylate and activate c-Src. In vivo overexpression of PTP1B in 293 cells caused a 2-fold increase of endogenous c-Src kinase activity. Our findings indicate that PTP1B is the primary protein-tyrosine phosphatase capable of dephosphorylating c-Src in several human breast cancer cell lines and suggests a regulatory role for PTP1B in the control of c-Src kinase activity. PMID- 11007775 TI - TatC is a specificity determinant for protein secretion via the twin-arginine translocation pathway. AB - The recent discovery of a ubiquitous translocation pathway, specifically required for proteins with a twin-arginine motif in their signal peptide, has focused interest on its membrane-bound components, one of which is known as TatC. Unlike most organisms of which the genome has been sequenced completely, the Gram positive eubacterium Bacillus subtilis contains two tatC-like genes denoted tatCd and tatCy. The corresponding TatCd and TatCy proteins have the potential to be involved in the translocation of 27 proteins with putative twin-arginine signal peptides of which approximately 6-14 are likely to be secreted into the growth medium. Using a proteomic approach, we show that PhoD of B. subtilis, a phosphodiesterase belonging to a novel protein family of which all known members are synthesized with typical twin-arginine signal peptides, is secreted via the twin-arginine translocation pathway. Strikingly, TatCd is of major importance for the secretion of PhoD, whereas TatCy is not required for this process. Thus, TatC appears to be a specificity determinant for protein secretion via the Tat pathway. Based on our observations, we hypothesize that the TatC-determined pathway specificity is based on specific interactions between TatC-like proteins and other pathway components, such as TatA, of which three paralogues are present in B. subtilis. PMID- 11007776 TI - A single dose of methamphetamine leads to a long term reversal of the blunted dopamine D1 receptor-mediated neocortical c-fos responses in mice deficient for D2 and D3 receptors. AB - Dopamine D(1) receptors play an essential role in the induction of expression of the immediate-early gene c-fos in response to pharmacological stimuli. In the forebrain of wild-type mice, administration of a D(1) receptor agonist leads to c fos mRNA expression levels that are substantially higher than corresponding levels expressed after indirect stimulation of dopamine receptors with methamphetamine. In mice deficient for D(2) and D(3) receptors, c-fos mRNA levels expressed in response to D(1) agonist administration are significantly blunted. However, a single dose of methamphetamine (5 mg/kg) leads to a long lasting reversal of the blunted c-fos responses in these mutants. In the forebrain, this reversal is restricted to the neocortex. Moreover, methamphetamine also enhances c-fos expression levels in preadolescent wild-type mice that normally express low c-fos mRNA in response to D(1) agonist stimulation. Thus, a single dose of methamphetamine leads to a long term increase in D(1) receptor-dependent c-fos responses in brains with either low (preadolescent mice) or blunted (adult D(2) and D(3) mutant mice) c-fos expression levels. A similar long term reversal of the blunted c-fos responses is achieved with a single dose of a full D(1) agonist. These results indicate that the constitutive inactivation of D(2) and D(3) receptors leads to a decrease in agonist-promoted D(1) receptor activity that can be reversed by intermittent agonist stimulation. PMID- 11007777 TI - Molecular cloning, expression, functional characterization, chromosomal localization, and gene structure of junctate, a novel integral calcium binding protein of sarco(endo)plasmic reticulum membrane. AB - Screening a cDNA library from human skeletal muscle and cardiac muscle with a cDNA probe derived from junctin led to the isolation of two groups of cDNA clones. The first group displayed a deduced amino acid sequence that is 84% identical to that of dog heart junctin, whereas the second group had a single open reading frame that encoded a polypeptide with a predicted mass of 33 kDa, whose first 78 NH(2)-terminal residues are identical to junctin whereas its COOH terminus domain is identical to aspartyl beta-hydroxylase, a member of the alpha ketoglutarate-dependent dioxygenase family. We named the latter amino acid sequence junctate. Northern blot analysis indicates that junctate is expressed in a variety of human tissues including heart, pancreas, brain, lung, liver, kidney, and skeletal muscle. Fluorescence in situ hybridization analysis revealed that the genetic loci of junctin and junctate map to the same cytogenetic band on human chromosome 8. Analysis of intron/exon boundaries of the genomic BAC clones demonstrate that junctin, junctate, and aspartyl beta-hydroxylase result from alternative splicing of the same gene. The predicted lumenal portion of junctate is enriched in negatively charged residues and is able to bind calcium. Scatchard analysis of equilibrium (45)Ca(2+) binding in the presence of a physiological concentration of KCl demonstrate that junctate binds 21.0 mol of Ca(2+)/mol protein with a k(D) of 217 +/- 20 microm (n = 5). Tagging recombinant junctate with green fluorescent protein and expressing the chimeric polypeptide in COS-7 transfected cells indicates that junctate is located in endoplasmic reticulum membranes and that its presence increases the peak amplitude and transient calcium released by activation of surface membrane receptors coupled to InsP(3) receptor activation. Our study shows that alternative splicing of the same gene generates the following functionally distinct proteins: an enzyme (aspartyl beta hydroxylase), a structural protein of SR (junctin), and a membrane-bound calcium binding protein (junctate). PMID- 11007778 TI - The enoyl-[acyl-carrier-protein] reductases FabI and FabL from Bacillus subtilis. AB - Enoyl-[acyl-carrier-protein] (ACP) reductase is a key enzyme in type II fatty acid synthases that catalyzes the last step in each elongation cycle. The FabI component of Bacillus subtilis (bsFabI) was identified in the genomic data base by homology to the Escherichia coli protein. bsFabI was cloned and purified and exhibited properties similar to those of E. coli FabI, including a marked preference for NADH over NADPH as a cofactor. Overexpression of the B. subtilis fabI gene complemented the temperature-sensitive growth phenotype of an E. coli fabI mutant. Triclosan was a slow-binding inhibitor of bsFabI and formed a stable bsFabI.NAD(+). triclosan ternary complex. Analysis of the B. subtilis genomic data base revealed a second open reading frame (ygaA) that was predicted to encode a protein with a relatively low overall similarity to FabI, but contained the Tyr-Xaa(6)-Lys enoyl-ACP reductase catalytic architecture. The purified YgaA protein catalyzed the NADPH-dependent reduction of trans-2-enoyl thioesters of both N-acetylcysteamine and ACP. YgaA was reversibly inhibited by triclosan, but did not form the stable ternary complex characteristic of the FabI proteins. Expression of YgaA complemented the fabI(ts) defect in E. coli and conferred complete triclosan resistance. Single knockouts of the ygaA or fabI gene in B. subtilis were viable, but double knockouts were not obtained. The fabI knockout was as sensitive as the wild-type strain to triclosan, whereas the ygaA knockout was 250-fold more sensitive to the drug. YgaA was renamed FabL to denote the discovery of a new family of proteins that carry out the enoyl-ACP reductase step in type II fatty-acid synthases. PMID- 11007780 TI - Targeting of Rac1 to the phagocyte membrane is sufficient for the induction of NADPH oxidase assembly. AB - The superoxide (O(2))-generating NADPH oxidase complex of phagocytes consists of a membrane-associated flavocytochrome (cytochrome b(559)) and four cytosolic proteins, p47(phox), p67(phox), p40(phox), and the small GTPase Rac (Rac1 or -2). NADPH oxidase activation (O(2) production) is elicited as the consequence of assembly of some or all cytosolic components with cytochrome b(559). This process can be reproduced in an in vitro system consisting of phagocyte membranes, p47(phox), p67(phox), and Rac, activated by an anionic amphiphile. We now show that post-translationally processed (prenylated) Rac1 initiates NADPH oxidase assembly, expressed in O(2) production, in a cell-free system containing phagocyte membrane vesicles and p67(phox), in the absence of an activating amphiphile and of p47(phox). Prenylated Cdc42Hs, a GTPase closely related to Rac, is inactive under the same conditions. Results obtained with phagocyte membrane vesicles can be reproduced fully by replacing these with partially purified cytochrome b(559), incorporated in phosphatidylcholine vesicles. Prenylated, but not nonprenylated, Rac1 binds spontaneously to phagocyte membrane vesicles and also to artificial, protein-free, phosphatidylcholine vesicles, a process counteracted by GDP dissociation inhibitor for Rho. Binding of prenylated Rac1 to membrane vesicles is accompanied by the recruitment of p67(phox) to the same location and the formation of an assembled NADPH oxidase complex, producing O(2) upon the addition of NADPH. Amphiphile and p47(phox)-independent NADPH oxidase activation by prenylated Rac1 is inhibited by Rho GDP dissociation inhibitor and by phosphatidylcholine vesicles, both competing with membrane for prenylated Rac1. We conclude that, in vitro, targeting of Rac to the phagocyte membrane is sufficient for the induction of NADPH oxidase assembly, suggesting that the principal or, possibly, the only role of Rac is to recruit cytosolic p67(phox) to the membrane environment, to be followed by the interaction of p67(phox) with cytochrome b(559). PMID- 11007779 TI - Calmodulin differentially modulates Smad1 and Smad2 signaling. AB - The members of the Smad protein family are intracellular mediators of transforming growth factor beta (TGF-beta) signaling. Smad1 transduces bone morphogenetic protein signals, inducing formation of ventral mesoderm in Xenopus embryos, whereas Smad2 transduces activin/TGF-beta signals, generating dorsal mesoderm. Calmodulin directly binds to many Smads and was shown to down-regulate Smad2 activity in a cell culture system (Zimmerman, C. M., Kariapper, M. S. T., and Mathews, L. S. (1997) J. Biol. Chem. 273, 677-680). Here, we extend those data and demonstrate that calmodulin alters Smad signaling in living embryos, increasing Smad1 activity while inhibiting Smad2 function. To characterize this regulation, we undertook a structure-function analysis and found that calmodulin binds to two distinct and conserved regions in both Smad1 and Smad2. Receptor tyrosine kinase signaling also modifies Smad activity (Kretzschmar, M., Doody, J., and Massague, J. (1997) Nature 389, 618-622; Kretzschmar, M., Doody, J., Timokhina, I., and Massague, J. (1999) Genes Dev. 13, 804-816; de Caestecker, M. P., Parks, W. T., Frank, C. J., Castagnino, P., Bottaro, D. P., Roberts, A. B., and Lechleider, R. J. (1998) Genes Dev. 12, 1587-1592). We show that calmodulin binding to Smads inhibits subsequent Erk2-dependent phosphorylation of Smads and vice versa. These observations suggest the presence of a cross-talk between three major signaling cascades as follows: Ca(2+)/calmodulin, receptor tyrosine kinase, and TGF-beta pathways. PMID- 11007781 TI - Radicicol binds and inhibits mammalian ATP citrate lyase. AB - Six different biotinylated radicicol derivatives were synthesized as affinity probes for identification of cellular radicicol-binding proteins. Derivatives biotinylated at the C-17 (BR-1) and C-11 (BR-6) positions retained the activity of morphological reversion in v-src-transformed 3Y1 fibroblasts. Two radicicol binding proteins, 120 and 90-kDa in size, were detected in HeLa cell extracts by employing BR-1 and BR-6, respectively. The 90-kDa protein bound to BR-6 was identified to be Hsp90 by immunoblotting. The 120-kDa protein bound to BR-1 was purified from rabbit reticulocyte lysate, and its internal amino acid sequence was identical to that of human and rat ATP citrate lyase. The identity of the 120 kDa protein as ATP citrate lyase was confirmed by immunoblotting. Interaction between BR-1 and ATP citrate lyase was blocked by radicicol but not by herbimycin A that interacts with Hsp90. These results suggest that radicicol binds the two proteins through different molecular portions of its structure. BR-1-bound ATP citrate lyase isolated from rabbit reticulocyte lysate showed no enzymatic activity. The activity of rat liver ATP citrate lyase was inhibited by radicicol and BR-1 but not by BR-6. Kinetic analysis demonstrated that radicicol was a non competitive inhibitor of ATP citrate lyase with K(i) values for citrate and ATP of 13 and 7 microm, respectively. PMID- 11007782 TI - Glycogen synthase kinase-3beta phosphorylates protein tau and rescues the axonopathy in the central nervous system of human four-repeat tau transgenic mice. AB - Protein tau filaments in brain of patients suffering from Alzheimer's disease, frontotemporal dementia, and other tauopathies consist of protein tau that is hyperphosphorylated. The responsible kinases operating in vivo in neurons still need to be identified. Here we demonstrate that glycogen synthase kinase-3beta (GSK-3beta) is an effective kinase for protein tau in cerebral neurons in vivo in adult GSK-3beta and GSK-3beta x human tau40 transgenic mice. Phosphorylated protein tau migrates slower during electrophoretic separation and is revealed by phosphorylation-dependent anti-tau antibodies in Western blot analysis. In addition, its capacity to bind to re-assembled paclitaxel (Taxol((R)))-stabilized microtubules is reduced, compared with protein tau isolated from mice not overexpressing GSK-3beta. Co-expression of GSK-3beta reduces the number of axonal dilations and alleviates the motoric impairment that was typical for single htau40 transgenic animals (Spittaels, K., Van den Haute, C., Van Dorpe, J., Bruynseels, K., Vandezande, K., Laenen, I., Geerts, H., Mercken, M., Sciot, R., Van Lommel, A., Loos, R., and Van Leuven, F. (1999) Am. J. Pathol. 155, 2153 2165). Although more hyperphosphorylated protein tau is available, neither an increase in insoluble protein tau aggregates nor the presence of paired helical filaments or tangles was observed. These findings could have therapeutic implications in the field of neurodegeneration, as discussed. PMID- 11007783 TI - Singlet oxygen induces oxidation of cellular DNA. AB - The aim of the present work was to evaluate the potential for (1)O(2) to induce oxidation of cellular DNA. For this purpose cells were incubated in the presence of a water-soluble endoperoxide whose thermal decomposition leads to the formation of singlet oxygen. Thereafter, DNA was extracted and the level of several modified DNA bases was determined by HPLC analysis coupled to a tandem mass spectrometric detection. A significant increase in the level of 8-oxo-7,8 dihydro-2'-deoxyguanosine was observed upon incubation of the cells with the chemical generator of (1)O(2), whereas the level of the other DNA bases measured remained unchanged. To demonstrate that singlet oxygen is directly involved in the formation of 8-oxo-7, 8-dihydro-2'-deoxyguanosine, the corresponding (18)O labeled endoperoxide was used. Incubation of the cells with such a generator of (18)O-labeled singlet oxygen results in the formation of (18)O-labeled 8-oxo-7,8 dihydro-2'-deoxyguanosine in the nuclear DNA. This result clearly demonstrates that singlet oxygen, when released within cells, is able to directly oxidize cellular DNA. PMID- 11007785 TI - The C-terminal polylysine region and methylation of K-Ras are critical for the interaction between K-Ras and microtubules. AB - After synthesis in the cytosol, Ras proteins must be targeted to the inner leaflet of the plasma membrane for biological activity. This targeting requires a series of C-terminal posttranslational modifications initiated by the addition of an isoprenoid lipid in a process termed prenylation. A search for factors involved in the intracellular trafficking of Ras has identified a specific and prenylation-dependent interaction between tubulin/microtubules and K-Ras. In this study, we examined the structural requirements for this interaction between K-Ras and microtubules. By using a series of chimeras in which regions of the C terminus of K-Ras were replaced with those of Ha-Ras and vice versa, we found that the polylysine region of K-Ras located immediately upstream of the prenylation site is required for binding of K-Ras to microtubules. Studies in intact cells confirmed the importance of the K-Ras polylysine region for microtubule binding, as deletion or replacement of this region resulted in loss of paclitaxel-induced mislocalization of a fluorescent K-Ras fusion protein. The additional modifications in the prenyl protein processing pathway also affected the interaction of K-Ras with microtubules. Removal of the three C-terminal amino acids of farnesylated K-Ras with the specific endoprotease Rce1p abolished its binding to microtubules. Interestingly, however, methylation of the C-terminal prenylcysteine restored binding. Consistent with these results, localization of the fluorescent K-Ras fusion protein remained paclitaxel-sensitive in cells lacking Rce1, whereas no paclitaxel effect was observed in cells lacking the methyltransferase. These studies show that the polylysine region of K-Ras is critical for its interaction with microtubules and provide the first evidence for a functional consequence of Ras C-terminal proteolysis and methylation. PMID- 11007784 TI - Ras inactivation of the retinoblastoma pathway by distinct mechanisms in NIH 3T3 fibroblast and RIE-1 epithelial cells. AB - Although Ras and Raf cause transformation of NIH 3T3 fibroblasts, only Ras causes transformation of RIE-1 intestinal epithelial cells. To determine if the inability of Raf to transform RIE-1 cells is due to a failure to deregulate cell cycle progression, we evaluated the consequences of sustained Ras and Raf activation on steady state levels of cyclin D1, p21(CIP/WAF), and p27(KIP1). Both Ras- and Raf-transformed NIH 3T3 cells showed up-regulated expression of cyclin D1, p21, and p27 protein, increased retinoblastoma (Rb) hyperphosphorylation, and increased activation of E2F-mediated transcription. Similarly, Ras-transformed RIE-1 cells also showed up-regulation of cyclin D1, p21, and hyperphosphorylated Rb. In contrast, Ras-mediated down-regulation of p27 was seen in RIE-1 cells. Conversely, stable expression of activated Raf alone caused only a partial up regulation of p21 and Rb hyperphosphorylation but no activation of E2F-responsive transcription or down-regulation of p27 in RIE-1 cells. Moreover, we found that the AP-1 site was dispensable for Ras-mediated stimulation of the cyclin-D1 promoter in NIH 3T3 cells but was essential for Ras-mediated stimulation in RIE-1 cells. Thus, up-regulation of p21, rather than the down-regulation seen in previous transient expression studies, is seen with sustained Ras activation. Additionally, p27 may serve a positive (NIH 3T3) or negative (RIE-1) regulatory role in Ras transformation that is cell type-dependent. The involvement of Raf and phosphatidylinositol 3-kinase in mediating Ras changes in cyclin D1, p21, and p27 was also very distinct in NIH 3T3 and RIE-1 cells. Taken together, these results demonstrate the importance of Raf-independent pathways in mediating oncogenic Ras deregulation of cell cycle progression in epithelial cells. PMID- 11007786 TI - Salivary agglutinin, which binds Streptococcus mutans and Helicobacter pylori, is the lung scavenger receptor cysteine-rich protein gp-340. AB - Salivary agglutinin is a high molecular mass component of human saliva that binds Streptococcus mutans, an oral bacterium implicated in dental caries. To study its protein sequence, we isolated the agglutinin from human parotid saliva. After trypsin digestion, a portion was analyzed by matrix-assisted laser/desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), which gave the molecular mass of 14 unique peptides. The remainder of the digest was subjected to high performance liquid chromatography, and the separated peptides were analyzed by MALDI-TOF/post-source decay; the spectra gave the sequences of five peptides. The molecular mass and peptide sequence information showed that salivary agglutinin peptides were identical to sequences in lung (lavage) gp-340, a member of the scavenger receptor cysteine-rich protein family. Immunoblotting with antibodies that specifically recognized either lung gp-340 or the agglutinin confirmed that the salivary agglutinin was gp-340. Immunoblotting with an antibody specific to the sialyl Le(x) carbohydrate epitope detected expression on the salivary but not the lung glycoprotein, possible evidence of different glycoforms. The salivary agglutinin also interacted with Helicobacter pylori, implicated in gastritis and peptic ulcer disease, Streptococcus agalactiae, implicated in neonatal meningitis, and several oral commensal streptococci. These results identify the salivary agglutinin as gp-340 and suggest it binds bacteria that are important determinants of either the oral ecology or systemic diseases. PMID- 11007787 TI - Coregulation of neurite outgrowth and cell survival by amphoterin and S100 proteins through receptor for advanced glycation end products (RAGE) activation. AB - Amphoterin is a protein enhancing process extension and migration in embryonic neurons and in tumor cells through binding to receptor for advanced glycation end products (RAGE), a multiligand transmembrane receptor. S100 proteins, especially S100B, are abundantly expressed in the nervous system and are suggested to function as cytokines with both neurotrophic and neurotoxic effects. However, the cell surface receptor for the cytokine function of S100B has not been identified. Here we show that two S100 family proteins, S100B and S100A1, activate RAGE in concert with amphoterin inducing neurite outgrowth and activation of transcription factor NF-kappaB. Furthermore, activation of RAGE by amphoterin and S100B promotes cell survival through increased expression of the anti-apoptotic protein Bcl-2. However, whereas nanomolar concentrations of S100B induce trophic effects in RAGE-expressing cells, micromolar concentrations of S100B induce apoptosis in an oxidant-dependent manner. Both trophic and toxic effects are specific for cells expressing full-length RAGE since cells expressing a cytoplasmic domain deletion mutant of RAGE are unresponsive to these stimuli. These findings suggest that activation of RAGE by multiple ligands is able to promote trophic effects whereas hyperactivation of RAGE signaling pathways promotes apoptosis. We suggest that RAGE is a signal-transducing receptor for both trophic and toxic effects of S100B. PMID- 11007788 TI - Altered distribution of the yeast plasma membrane H+-ATPase as a feature of vacuolar H+-ATPase null mutants. AB - The effect of vacuolar H(+)-ATPase (V-ATPase) null mutations on the targeting of the plasma membrane H(+)-ATPase (Pma1p) through the secretory pathway was analyzed. Gas1p, which is another plasma membrane component, was used as a control for the experiments with Pma1p. Contrary to Gas1p, which is not affected by the deletion of the V-ATPase complex in the V-ATPase null mutants, the amount of Pma1p in the plasma membrane is markedly reduced, and there is a large accumulation of the protein in the endoplasmic reticulum. Kex2p and Gef1p, which are considered to reside in the post-Golgi vesicles, were suggested as required for the V-ATPase function; hence, their null mutant phenotype should have been similar to the V-ATPase null mutants. We show that, in addition to the known differences between those yeast phenotypes, deletions of KEX2 or GEF1 in yeast do not affect the distribution of Pma1p as the V-ATPase null mutant does. The possible location of the vital site of acidification by V-ATPase along the secretory pathway is discussed. PMID- 11007789 TI - The enigma of cobalamin (Vitamin B12) biosynthesis in Porphyromonas gingivalis. Identification and characterization of a functional corrin pathway. AB - The ability of Porphyromonas gingivalis to biosynthesize tetrapyrroles de novo has been investigated. Extracts of the bacterium do not possess activity for 5- aminolevulinic-acid dehydratase or porphobilinogen deaminase, two key enzymes involved in the synthesis of uroporphyrinogen III. Similarly, it was not possible to detect any genetic evidence for these early enzymes with the use of degenerate polymerase chain reaction. However, the bacterium does appear to harbor some of the enzymes for cobalamin biosynthesis since cobyric acid, a pathway intermediate, was converted into cobinamide. Furthermore, degenerate polymerase chain reaction with primers to cbiP, which encodes cobyric-acid synthase, produced a fragment with a high degree of identity to Salmonella typhimurium cbiP. Indeed, the recently released genome sequence data confirmed the presence of cbiP together with 14 other genes of the cobalamin pathway. A number of these genes were cloned and functionally characterized. Although P. gingivalis harbors all the genes necessary to convert precorrin-2 into cobalamin, it is missing the genes for the synthesis of precorrin-2. Either the organism has a novel pathway for the synthesis of precorrin-2, or more likely, it has lost this early part of the pathway. The remainder of the pathway may be being maintained to act as a salvage route for corrin synthesis. PMID- 11007790 TI - High glucose inhibits glucose-6-phosphate dehydrogenase via cAMP in aortic endothelial cells. AB - Recent studies have shown that hyperglycemia is a principal cause of cellular damage in patients with diabetes mellitus. A major consequence of hyperglycemia is increased oxidative stress. Glucose-6-phosphate dehydrogenase (G6PD) plays an essential role in the regulation of oxidative stress by primarily regulating NADPH, the main intracellular reductant. In this paper we show that increased glucose (10-25 mm) caused inhibition of G6PD resulting in decreased NADPH levels in bovine aortic endothelial cells (BAEC). Inhibition was seen within 15 min. High glucose-induced inhibition of G6PD predisposed cells to cell death. High glucose via increased activity of adenylate cyclase also stimulated an increase in cAMP levels in BAEC. Agents that increased cAMP caused a decrease in G6PD activity. Inhibition of cAMP-dependent protein kinase A ameliorated the high glucose-induced inhibition of G6PD. Finally, high glucose stimulated phosphorylation of G6PD. These results suggest that, in BAEC, high glucose stimulated increased cAMP, which led to increased protein kinase A activity, phosphorylation of G6PD, and inhibition of G6PD activity. We conclude that these changes in G6PD activity play an important role in high glucose-induced cell damage/death. PMID- 11007791 TI - The crystal structure of 3alpha -hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni shows a novel oligomerization pattern within the short chain dehydrogenase/reductase family. AB - The crystal structure of 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni (3alpha-HSDH) as well as the structure of its binary complex with NAD(+) have been solved at 1.68-A and 1.95-A resolution, respectively. The enzyme is a member of the short chain dehydrogenase/reductase (SDR) family. Accordingly, the active center and the conformation of the bound nucleotide cofactor closely resemble those of other SDRs. The crystal structure reveals one homodimer per asymmetric unit representing the physiologically active unity. Dimerization takes place via an interface essentially built-up by helix alphaG and strand betaG of each subunit. So far this type of intermolecular contact has exclusively been observed in homotetrameric SDRs but never in the structure of a homodimeric SDR. The formation of a tetramer is blocked in 3alpha HSDH by the presence of a predominantly alpha-helical subdomain which is missing in all other SDRs of known structure. PMID- 11007792 TI - Detection of large pKa perturbations of an inhibitor and a catalytic group at an enzyme active site, a mechanistic basis for catalytic power of many enzymes. AB - Delta(5)-3-Ketosteroid isomerase catalyzes cleavage and formation of a C-H bond at a diffusion-controlled limit. By determining the crystal structures of the enzyme in complex with each of three different inhibitors and by nuclear magnetic resonance (NMR) spectroscopic investigation, we evidenced the ionization of a hydroxyl group (pK(a) approximately 16.5) of an inhibitor, which forms a low barrier hydrogen bond (LBHB) with a catalytic residue Tyr(14) (pK(a) approximately 11.5), and the protonation of the catalytic residue Asp(38) with pK(a) of approximately 4.5 at pH 6.7 in the interaction with a carboxylate group of an inhibitor. The perturbation of the pK(a) values in both cases arises from the formation of favorable interactions between inhibitors and catalytic residues. The results indicate that the pK(a) difference between catalytic residue and substrate can be significantly reduced in the active site environment as a result of the formation of energetically favorable interactions during the course of enzyme reactions. The reduction in the pK(a) difference should facilitate the abstraction of a proton and thereby eliminate a large fraction of activation energy in general acid/base enzyme reactions. The pK(a) perturbation provides a mechanistic ground for the fast reactivity of many enzymes and for the understanding of how some enzymes are able to extract a proton from a C-H group with a pK(a) value as high as approximately 30. PMID- 11007793 TI - Characterization of variants altered at the N-terminal proline, a novel heme axial ligand in CooA, the CO-sensing transcriptional activator. AB - CooA, the carbon monoxide-sensing transcription factor from Rhodospirillum rubrum, binds CO through a heme moiety resulting in conformational changes that promote DNA binding. The crystal structure shows that the N-terminal Pro(2) of one subunit (Met(1) is removed post-translationally) provides one ligand to the heme of the other subunit in the CooA homodimer. To determine the importance of this novel ligand and the contiguous residues to CooA function, we have altered the N terminus through two approaches: site-directed mutagenesis and regional randomization, and characterized the resulting CooA variants. While Pro(2) appears to be optimal for CooA function, it is not essential and a variety of studied variants at this position have substantial CO-sensing function. Surprisingly, even alterations that add a residue (where Pro(2) is replaced by Met(1)-Tyr(2), for example) accumulate heme-containing CooA with functional properties that are similar to those of wild-type CooA. Other nearby residues, such as Phe(5) and Asn(6) appear to be important for either the structural integrity or the function of CooA. These results are contrasted with those previously reported for alteration of the His(77) ligand on the opposite side of the heme. PMID- 11007794 TI - Interaction between 4-hydroxy-2,3-alkenals and the platelet-derived growth factor beta receptor. Reduced tyrosine phosphorylation and downstream signaling in hepatic stellate cells. AB - Hepatic stellate cells (HSC) undergo activation toward myofibroblast-like cells during early stages of liver injury associated with fibrogenesis. Platelet derived growth factor (PDGF), particularly its BB isoform, has been identified as the most potent mitogen for HSC. 4-Hydroxy-2,3-nonenal and related 4-hydroxy-2, 3 alkenals (HAKs) have been suggested to modulate the process of HSC activation. In this study we investigated the relationship between HAKs and PDGF receptor activation in human HSC. By employing noncytotoxic concentrations (10(-6) m) of HAKs, we observed a significant inhibition of PDGF-BB-dependent DNA synthesis. HAKs inhibited relevant pathways of PDGF-BB-dependent mitogenic signaling, including autophosphorylation of PDGF receptor (PDGF-R) beta subunits and activation of phosphatidylinositol 3-kinase and extracellular regulated kinases 1/2. Inhibition of DNA synthesis was reversible, and recovery of PDGF-mediated mitogenic signaling occurred within 24-48 h and was associated with HAKs-induced up-regulation of PDGF-R beta gene expression. 4-Hydroxy-2,3-nonenal, used as a model HAK, inhibited the intrinsic tyrosine kinase activity associated with the PDGF-R beta subunit, whereas binding of PDGF to its receptor was unaffected. This study identifies a novel regulatory mechanism of reactive aldehydes on PDGF receptor signaling and biologic actions, which may be relevant in several pathophysiological conditions, including liver fibrosis. PMID- 11007795 TI - The amino-terminal part of PRELP binds to heparin and heparan sulfate. AB - PRELP (proline, arginine-rich end leucine-rich repeat protein) is an extracellular matrix leucine-rich repeat protein. The amino-terminal region of PRELP differs from that of other leucine-rich repeat proteins in containing a high number of proline and arginine residues. The clustered proline and basic residues are conserved in rat, bovine, and human PRELP. Although the function of PRELP is not yet known, the clustered arginine residues suggest a heparan sulfate/heparin-binding capacity. We show here that PRELP indeed binds heparin and heparan sulfate. Truncated PRELP without the amino-terminal region does not bind heparin. The dissociation constant for the interaction of PRELP with heparin was determined by an in solution binding assay and by surface plasmon resonance analysis to be in the range of 10-30 nm. A 6-mer heparin oligosaccharide was the smallest size showing binding to PRELP. The binding increased with increasing length up to an 18-mer and depended on the degree of sulfation of heparin as well as heparan sulfate. Sulfate groups at all positions were shown to be of importance for the binding. Fibroblasts bind PRELP, and this interaction is inhibited with heparin, suggesting a function for PRELP as a linker between the matrix and cell surface proteoglycans. PMID- 11007796 TI - Glucose activates mitogen-activated protein kinase (extracellular signal regulated kinase) through proline-rich tyrosine kinase-2 and the Glut1 glucose transporter. AB - Glucose serves as both a nutrient and regulator of physiological and pathological processes. Presently, we found that glucose and certain sugars rapidly activated extracellular signal-regulated kinase (ERK) by a mechanism that was: (a) independent of glucose uptake/metabolism and protein kinase C but nevertheless cytochalasin B-inhibitable; (b) dependent upon proline-rich tyrosine kinase-2 (PYK2), GRB2, SOS, RAS, RAF, and MEK1; and (c) amplified by overexpression of the Glut1, but not Glut2, Glut3, or Glut4, glucose transporter. This amplifying effect was independent of glucose uptake but dependent on residues 463-468, IASGFR, in the Glut1 C terminus. Accordingly, glucose effects on ERK were amplified by expression of Glut4/Glut1 or Glut2/Glut1 chimeras containing IASGFR but not by Glut1/Glut4 or Glut1/Glut2 chimeras lacking these residues. Also, deletion of Glut1 residues 469-492 was without effect, but mutations involving serine 465 or arginine 468 yielded dominant-negative forms that inhibited glucose dependent ERK activation. Glucose stimulated the phosphorylation of tyrosine residues 402 and 881 in PYK2 and binding of PYK2 to Myc-Glut1. Our findings suggest that: (a) glucose activates the GRB2/SOS/RAS/RAF/MEK1/ERK pathway by a mechanism that requires PYK2 and residues 463-468, IASGFR, in the Glut1 C terminus and (b) Glut1 serves as a sensor, transducer, and amplifier for glucose signaling to PYK2 and ERK. PMID- 11007797 TI - The acceptor and site specificity of alpha 3-fucosyltransferase V. High reactivity of the proximal and low of the distal galbeta 1-4GlcNAc unit in i-type polylactosamines. AB - We report here on in vitro acceptor and site specificity of recombinant alpha3 fucosyltransferase V (Fuc-TV) with 40 oligosaccharide acceptors. Galbeta1-4GlcNAc (LN) and GalNAcbeta1-4GlcNAc (LDN) reacted rapidly; Galbeta1-3GlcNAc (LNB) reacted moderately, and GlcNAcbeta1-4GlcNAc (N, N'-diacetyl-chitobiose) reacted slowly yet distinctly. In neutral and terminally alpha3-sialylated polylactosamines of i-type, the reducing end LN unit reacted rapidly and the distal (sialyl)LN group very slowly; the midchain LNs revealed intermediate reactivities. The data suggest that a distal LN neighbor enhances but a proximal LN neighbor reduces the reactivity of the midchain LNs. This implies that Fuc-TV may bind preferably the tetrasaccharide sequence Galbeta1-4GlcNAcbeta1-3Galbeta1 4GlcNAc for transfer at the underlined monosaccharide. Terminal alpha3 sialylation of i-type polylactosamines almost doubled the reactivities of the LN units at all positions of the chains. We conclude that, in comparison with human Fuc-TIV and Fuc-TIX, Fuc-TV reacted with a highly distinct site specificity with i-type polylactosamines. The Fuc-TV reactivity of free LNB resembled that of LNBbeta1-3'R of a polylactosamine, contrasting strongly with the dissimilarity of the reactivities of the analogous pair of LN and LNbeta1-3'R. This observation supports the notion that LN and LNB may be functionally bound at distinct sites on Fuc-TV surface. Our data show that Fuc-TV worked well with a very wide range of LN-glycans, showing weak reactivity only with distal (sialyl)LN units of i type polylactosamines, biantennary N-glycans, and I branches of polylactosamines. PMID- 11007798 TI - Localization of the death domain of tissue inhibitor of metalloproteinase-3 to the N terminus. Metalloproteinase inhibition is associated with proapoptotic activity. AB - The tissue inhibitors of metalloproteinases (TIMPs) are a family of four secreted inhibitors of matrix metalloproteinases (MMPs). Recently, additional functions have been attributed to the TIMPs, including cell growth and inhibition of angiogenesis. In particular, we demonstrated that TIMP-3 overexpression using gene transfer induces apoptosis in a variety of cell types and can inhibit vascular neointima formation in vivo. However, little is know about the mechanisms underlying TIMP-3-mediated apoptosis. Here, using both purified recombinant proteins and novel adenoviral vectors we demonstrate that the prodeath domain of TIMP-3 is located within the N-terminal three loops of TIMP-3. Although both wild type and N-terminal TIMP-3 proteins promoted apoptosis, a T 2/T-3 chimera, in which the N-terminal three loops of TIMP-3 are replaced by those of TIMP-2, failed to induce cell death. Furthermore, a point mutation at residue 1 of TIMP-3 totally abolished MMP-inhibitory activity of TIMP-3 and also failed to promote apoptosis. This study demonstrates, using multiple apoptosis assays, that the prodeath function of TIMP-3 is located within the N-terminal three loops and the presence of functional metalloproteinase-inhibitory activity is associated with the induction of apoptosis. PMID- 11007799 TI - cDNA cloning and expression of a human aldehyde dehydrogenase (ALDH) active with 9-cis-retinal and identification of a rat ortholog, ALDH12. AB - This report describes the isolation of a heretofore uncharacterized aldehyde dehydrogenase (ALDH) with retinal dehydrogenase activity from rat kidney and the cloning and expression of a cDNA that encodes its human ortholog, the previously unknown ALDH12. The human ALDH12 cDNA predicts a 487-residue protein with the 23 invariant amino acids, four conserved regions, cofactor binding motif (G(209)XGX(3)G), and active site cysteine residue (Cys(287)) that typify members of the ALDH superfamily. ALDH12 seems at least as efficient (V(m)/K(m)) in converting 9-cis-retinal into the retinoid X receptor ligand 9-cis-retinoic acid as two previously identified ALDHs with 9-cis-retinal dehydrogenase activity, rat retinal dehydrogenase (RALDH) 1 and RALDH2. ALDH12, however, has approximately 40 fold higher activity with 9-cis- retinal than with all-trans-retinal, whereas RALDH1 and RALDH2 have equivalent and approximately 4-fold less efficiencies for 9-cis-retinal versus all-trans-retinal, respectively. Therefore, ALDH12 is the first known ALDH to show a preference for 9-cis-retinal relative to all-trans retinal. Evidence consistent with the possibility that ALDH12 could function in a pathway of 9-cis-retinoic acid biosynthesis in vivo includes biosynthesis of 9 cis-retinoic acid from 9-cis-retinol in cells co-transfected with cDNAs encoding ALDH12 and the 9-cis-retinol/androgen dehydrogenase, cis-retinoid/androgen dehydrogenase type 1. Intense ALDH12 mRNA expression in adult and fetal liver and kidney, two organs that reportedly have relatively high concentrations of 9-cis retinol, reinforces this notion. PMID- 11007800 TI - p53 amino acids 339-346 represent the minimal p53 repression domain. AB - The p53 tumor suppressor protein functions as an activator and also as a repressor of gene transcription. Currently, the mechanism of transcriptional repression by p53 remains poorly understood. To help clarify this mechanism, we carried out studies designed to identify the minimal repression domain that inhibits p53 transcriptional activities. We found only eight amino acids (339) of the COOH-terminal domain (termed P53MRD) that possess activities of repression. The exact location of this minimal domain is on the E6-binding region, and it lacks the ability of tetramerization. P53MRD is able to repress the transcription of p53 while not affecting VP16. The mutants (amino acids M340P and F341D) of native p53 also lost transcriptional repression of the thymidine kinase chloramphenicol acetyltransferase promoter. These results suggest that this eight amino acid element is required for the repression of p53. PMID- 11007801 TI - Huntingtin interacting protein 1 induces apoptosis via a novel caspase-dependent death effector domain. AB - Huntington disease is a devastating neurodegenerative disease caused by the expansion of a polymorphic glutamine tract in huntingtin. The huntingtin interacting protein (HIP-1) was identified by its altered interaction with mutant huntingtin. However, the function of HIP-1 was not known. In this study, we identify HIP-1 as a proapoptotic protein. Overexpression of HIP-1 resulted in rapid caspase 3-dependent cell death. Bioinformatics analyses identified a novel domain in HIP-1 with homology to death effector domains (DEDs) present in proteins involved in apoptosis. Expression of the HIP-1 DED alone resulted in cell death indistinguishable from HIP-1, indicating that the DED is responsible for HIP-1 toxicity. Furthermore, substitution of a conserved hydrophobic phenylalanine residue within the HIP-1 DED at position 398 eliminated HIP-1 toxicity entirely. HIP-1 activity was found to be independent of the DED containing caspase 8 but was significantly inhibited by the antiapoptotic protein Bcl-x(L), implicating the intrinsic pathway of apoptosis in HIP-1-induced cell death. Co-expression of a normal huntingtin fragment capable of binding HIP-1 significantly reduced cell death. Our data identify HIP-1 as a novel proapoptotic mediator and suggest that HIP-1 may be a molecular accomplice in the pathogenesis of Huntington disease. PMID- 11007803 TI - Journal of pediatric psychology: A brief history (1969-1999) PMID- 11007802 TI - Glucose residues as key determinants in the biosynthesis and quality control of glycoproteins with N-linked oligosaccharides. PMID- 11007804 TI - Family rituals as a protective factor for children with asthma. AB - OBJECTIVE: To examine how one aspect of family life, notably family rituals and routines, may protect children with asthma from anxiety-related symptoms. METHODS: Eighty-six families (43 children with asthma, 43 healthy comparison children) participated in the study. Children completed measures of anxiety (Revised Child Manifest Anxiety Scale) and health. Parents completed measures of stress (Parenting Stress Index), family rituals (Family Routines Questionnaire), and family health. RESULTS: Families that reported more meaning in their family routines had children who reported lower levels of anxiety. Mother endorsement of family ritual meaning and father endorsement of family ritual routine were most strongly related to lower levels of anxiety. Support for the protective function of meaningful family rituals was stronger when a general health stress model was used rather than the presence or absence of asthma alone. CONCLUSIONS: Family rituals may serve a protective function for children with asthma under conditions of heightened parenting stress. PMID- 11007805 TI - The influence of parent-child relatedness on depressive symptoms in children with asthma: tests of moderator and mediator models. AB - OBJECTIVE: To examine the influence of parent-child relationship quality on the association between illness-related functional status and depressive symptoms in children with asthma. METHODS: Questionnaire data were collected from the child, caregiver, and physician. Fifty-five children with asthma (8-17 years of age), their caregivers, and physicians participated. RESULTS: Regression analyses suggest that patterns of mother-child relatedness (secure vs. insecure) mediate the relationship between functional status and depressive symptoms. CONCLUSIONS: The parent-child relationship may be an important pathway by which illness influences symptoms of depression in children with asthma. This study suggests that impaired functional status does not directly contribute to symptoms of depression, but rather influences the parent-child relationship in ways that may promote the development of depressive symptoms in the child. PMID- 11007806 TI - Aggression, antisocial behavior, and substance abuse in survivors of pediatric cancer: possible protective effects of cancer and its treatment. AB - OBJECTIVE: To examine aggression, antisocial behavior, and substance abuse in young adult survivors of pediatric cancer (PCS) relative to case control peers (CC). METHODS: We obtained self-reports of current aggression, antisocial behavior, and lifetime substance use from 26 PCS (time off-treatment, M: = 56 months) and 26 CC using the Antisocial Behavior Checklist and the Drinking and Drug History. A report of current aggression and antisocial behavior also was obtained from primary caregivers using the Child Behavior Checklist. RESULTS: PCS self-reported significantly less illegal drug use and experimentation than CC. No significant differences emerged between groups for use of alcohol and tobacco nor for aggression and antisocial behavior. CONCLUSIONS: PCS are functioning as well as, or better than, CC in terms of aggression, antisocial behavior, and substance abuse. However, given the compromised health status of survivors, efforts should focus on further reduction of drug-related risk behaviors that may amplify organ damage or increase risk for further malignancies in this population. PMID- 11007807 TI - Neurocognitive functioning and magnetic resonance imaging in children with sickle cell disease. AB - OBJECTIVE: To examine neurocognitive functioning in children classified with overt cerebral vascular accidents (CVAs), silent infarcts, or without central nervous system (CNS) pathology on magnetic resonance imaging. METHODS: Participants were 63 children and adolescents with sickle cell disease (SCD). RESULTS: Children with overt CVAs and silent infarcts differed from their peers without CNS pathology on measures of attention and executive functioning. CONCLUSIONS: We consider these deficits the result of the high frequency of frontal lobe deficits incurred by children with SCD. Recommendations include the use of tests designed to measure attention and executive functioning as a way of screening children with SCD for possible CNS pathology. We also suggest that future research examine the mechanism underlying frontal lobe involvement for individuals with SCD. PMID- 11007808 TI - Brief report: speed of information processing in children with insulin-dependent diabetes mellitus. AB - OBJECTIVE: To determine whether children with insulin-dependent diabetes mellitus (IDDM) process information more slowly than children who do not have diabetes. METHODS: We tested 31 children with early onset and longer duration of IDDM, 35 with later onset and briefer duration of IDDM, and 36 comparison children without diabetes. They were administered five tasks requiring rapid responding that assessed a range of cognitive processes. RESULTS: On most tasks, children in the three groups were quite similar in the accuracy and speed of performance. Furthermore, for children in the diabetic groups, disease-related variables were unrelated to accuracy and speed of performance. CONCLUSIONS: The results suggest that children with IDDM do not have a pervasive deficit in speed of information processing, although more circumscribed deficits in processing speed are possible. PMID- 11007809 TI - Pioneers in pediatric psychology: a career in pediatric psychology. PMID- 11007810 TI - Microvascular disease--the Cinderella of uraemic heart disease. AB - It has been known for a long time that atherosclerosis, particularly plaques in the epicardiac coronary conduit arteries, are more frequent in patients with chronic renal failure than in non-uraemic patients. It has been only recently, however, that modification of post-stenotic remodelling of cardiac arteries as well as abnormalities of the arterioles and the capillaries in the myocardium of uraemic animals and uraemic patients have been recognized and analysed. These lesions can be dissociated from changes in blood pressure and may be an important cause contributing to reduced ischaemia tolerance and cardiac malfunction (pump failure, arrhythmia) thus predisposing to cardiac death. Recent insights into angiogenesis, particularly adaptive angiogenesis in response to hypoxia, may potentially provide novel approaches to the understanding and management of cardiac microangiopathy in renal failure. PMID- 11007811 TI - Macula densa signalling--a potential role of cyclooxygenase-2 (COX-2)? PMID- 11007812 TI - Is there a role for locally produced complement in renal disease? PMID- 11007813 TI - Smoking and the kidney. PMID- 11007814 TI - Pastis and hypertension--what is the molecular basis? PMID- 11007815 TI - Hypertensive nephrosclerosis-a cause of end-stage renal disease? PMID- 11007816 TI - Acute renal failure related to the crush syndrome: towards an era of seismo nephrology? PMID- 11007817 TI - Malnutrition and the acute-phase reaction in dialysis patients-how to measure and how to distinguish. PMID- 11007818 TI - End-stage renal disease and erectile dysfunction. Is there any hope? PMID- 11007819 TI - The management of chronic renal insufficiency in the conservative phase. PMID- 11007820 TI - Effects of pentoxifylline, pentifylline and gamma-interferon on proliferation, differentiation, and matrix synthesis of human renal fibroblasts. AB - BACKGROUND: Kidneys that progress to end-stage renal failure are almost invariably characterized by the presence of tubulointerstitial fibrosis. Therapeutic interventions to halt the progressive deterioration of renal function are still limited. Pentoxifylline, pentifylline, and gamma-interferon have shown a potential benefit in the treatment of fibrotic processes in the skin and lung. Thus, the aim of the present study was the analysis of potential anti-fibrotic effects of these substances on human kidney fibroblasts in vitro. METHODS: Primary renal fibroblasts were established from human kidney biopsies and were studied in addition to two renal fibroblast cell lines. Cells were first growth arrested by withdrawal of fetal calf serum (FCS) and subsequently stimulated with 10% FCS in the presence of different concentrations of pentoxifylline (PTX), pentifylline (PTF), or gamma-interferon (IFN-gamma). Fibroblast proliferation was determined by bromodeoxyuridine incorporation and cell counts. Northern and western blot hybridizations for basic fibroblast growth factor (FGF)-2 and transforming growth factor (TGF)-beta1 were performed to analyse inhibitory effects. The effects of all three substances on matrix synthesis were evaluated by immunoblot analyses and ELISA for collagen type I and fibronectin after stimulation with TGF-beta1. Finally, differentiation into myofibroblasts was examined by double immunofluorescence staining for alpha-smooth-muscle actin and Hoechst dye H33258. RESULTS: PTX and PTF resulted in a dose- and time-dependent inhibition of proliferation in all fibroblast lines (maximum 78.9+/-6.2% at 500 microg/ml PTX). Conversely, IFN-gamma had only modest effects on fibroblast proliferation, resulting in a maximum of 36.0+/-6.1% inhibition at 500 U/ml. Northern blot hybridizations determined that FGF-2 mRNA levels in fibroblasts were decreased up to 73.7 and 91.5% by PTX (1000 microg/ml) and PTF (100 microg/ml), whereas IFN-gamma led to a reduction of 46.2% at 1000 U/ml, indicating that the inhibitory effects of all three substances may be mediated through inhibition of FGF-2 synthesis. These findings were corroborated by immunoblot analyses where again PTX and PTF had the strongest inhibitory effects. No change in TGF-beta1 mRNA levels was noted. Synthesis of cellular and secreted collagen type I was robustly inhibited by PTX and PTF, whereas IFN-gamma exerted the strongest inhibitory effect on fibronectin synthesis and secretion. In addition, IFN-gamma down-regulated the expression of alpha-smooth-muscle actin up to 73.3% (at 1000 U/ml) whereas PTX and PTF resulted in a down-regulation of up to 49.7+/-1.8 and 80.0+/-4.4% (at 1000 and 100 microg/ml) respectively. PTF was in all experiments about 10 times more potent than equimolar concentrations of PTX. CONCLUSIONS: PTX and PTF exerted robust inhibitory effects on fibroblast proliferation, extracellular matrix synthesis, and myofibroblastic differentiation. Conversely, IFN-gamma caused strong inhibition of fibronectin synthesis and alpha-smooth-muscle cell actin expression but had only weak inhibitory influences on fibroblast proliferation and collagen type I synthesis. Inhibitory effects of all three substances on proliferation may be mediated through inhibition of FGF-2 synthesis. PMID- 11007821 TI - Glomerular function and morphology in puromycin aminonucleoside nephropathy in rats. AB - BACKGROUND: The most characteristic manifestation of minimal-change nephropathy is podocyte cell process broadening. In a previous study in children from our unit, we found an inverse correlation between foot process width, glomerular filtration rate (GFR), and filtration fraction. The aim of the present study was to determine whether this relationship also existed in the puromycin aminonucleoside (PAN) experimental model. METHODS: Sixteen Munich-Wistar-Fromter male rats initially weighing median 247 g (range 171-286) were used. Four rats served as controls. The other 12 rats were divided into three groups receiving daily subcutaneous injections of 1, 1.67, and 2.5 mg PAN/100 g body weight respectively, for 6 days. GFR was determined by clearance of inulin and the fractional urine albumin excretion was measured. Standard stereological methods were used to estimate the glomerular volume, the mean foot process width and the length density of slit pores. RESULTS: GFR decreased with increasing PAN doses. The glomerular volume was increased in the group receiving the lowest PAN dose, while it was decreased in the group with the highest PAN dose, compared with controls. The fractional albumin excretion and the foot process width increased and the total slit pore length decreased with increasing doses of PAN. GFR correlated directly with the glomerular volume as did the foot process width with the fractional albumin excretion. The foot process width correlated inversely with the glomerular volume as did the glomerular volume with the fractional albumin excretion, and GFR with foot process width. CONCLUSIONS: The decreased GFR found in the nephrotic rats was inversely related to foot process width and directly related to glomerular volume, confirming our previous results in children in an early stage of the nephrotic syndrome. PMID- 11007822 TI - Antibody-induced modulation of the leukocyte CD11b integrin prevents mild but not major renal ischaemic injury. AB - BACKGROUND: CD11/CD18 beta(2) integrins are involved in leukocyte adhesion to the activated endothelium, and therefore represent a possible therapeutic target in the prevention of ischaemic acute renal failure (ARF). METHODS: To assess the effect of an anti-CD11b monoclonal antibody (mAb) in ischaemic ARF, uninephrectomized Fischer rats were subjected to 45 or 60 min of warm renal ischaemia, then received 1 mg of anti-CD11b mAb 5 min before reperfusion. RESULTS: After 45 min of ischaemia, renal function tests at 24 and 48 h were less altered in mAb-treated than in control rats, but after 60 min of ischaemia the same level of renal insufficiency was observed in the two groups. In parallel, milder tubular necrosis and less leukocyte infiltration were observed in the treated group after 45 min of ischaemia, but no difference was seen after 60 min compared to the control group. The mAb was detected on blood neutrophils up to 48 h after infusion and a marked down-regulation of CD11b expression on neutrophil surfaces was documented by flow cytometry. CONCLUSION: These results indicate that anti-CD11b mAb administered prior to reperfusion decreases moderate ischaemic ARF but fails to prevent renal injury secondary to prolonged ischaemia in this model. PMID- 11007823 TI - Identification and kinetics of leukocytes after severe ischaemia/reperfusion renal injury. AB - BACKGROUND: Leukocyte adhesion/infiltration in response to renal ischaemia/reperfusion (I/R) injury is a well-known but poorly understood phenomenon. The identification, kinetics, and exact role of these inflammatory cells in I/R injury and regeneration are still matters of debate. METHODS: Uninephrectomized rats were submitted to 60 min renal ischaemia by clamping of renal vessels. RESULTS: Severe acute renal failure was observed, with maximum functional impairment on day 2. By 12 h after the ischaemic event, up to 80% of proximal tubular cells in the outer stripe of outer medulla (OSOM) were already severely damaged. Proliferation (proliferating cell nuclear antigen (PCNA) staining) started after 24 h, reaching maximum activity on day 3. Regeneration of tubular morphology started on the 3rd day, and after 10 days 50% of tubules had regenerated completely. Interstitial leukocytes (OX-1 immunohistochemical staining) were already prominent at day 1, thereafter gradually increasing with time. The so-called neutrophil-specific identification methods (myeloperoxidase (MPO), chloroacetate esterase, mAb HIS-48) proved to be non-specific, since they also stained for macrophages, as demonstrated by flow cytometry and the combination of these stainings with the macrophage-specific ED-1 staining. MPO activity was already significantly increased at 1 h post-I/R (439+/-34%, P<0.005), reaching its maximum activity after 12 h of I/R (1159+/-138%, P<0.0005), declining thereafter. On the other hand, neutrophil presence investigated by H&E staining revealed only a few neutrophils in glomeruli, medullary rays, and OSOM at 24 h after the ischaemic event (4.7+/-4.2 cells/mm(2) vs controls=2.3+/-2.0 cells/mm(2) (n.s.)), and remained unchanged over the next 10 days. In contrast, significant monocyte/macrophage adhesion/infiltration (ED-1 staining) occurred at the OSOM at 24 h post-ischaemia (at 24 h, 120+/-46 cells/mm(2) vs. sham=18+/-4 cells/mm(2) (P<0.05)), became prominent at day 5 (1034+/-161 cells/mm(2) vs sham=18+/-18 cells/mm(2) (P<0.05)), and almost disappeared after 10 days. CD4(+) cells (W3/25) gradually increased from day 5, reaching a maximum at day 10. A few CD8(+) cells (OX-8) were apparent from days 3 until 10, but no B-cells (OX-33) were observed. CONCLUSIONS: After severe warm I/R renal injury, a pronounced acute tubular necrosis occurs during the first 12 24 h in the absence of a marked cellular infiltrate, but with an important renal MPO activity, reflecting the activation of the adhering inflammatory cells (polymorphonuclear cells (PMNs) and mainly monocytes/macrophages). Only later at the time and site (OSOM) of regeneration a sequential accumulation of monocytes/macrophages and T cells becomes prominent, in contrast with the low number of neutrophils found in the kidney during the 10-day post-ischaemic period. The non-specificity of the so-called neutrophil-specific identification methods (MPO activity, naphthol AS-D chloroacetate esterase, or mAb HIS-48 staining), cross-reacting with monocytes/macrophages, explains the controversy in literature concerning the number of PMNs in post-ischaemic injury. PMID- 11007824 TI - Renal function of children exposed to cyclosporin in utero. AB - BACKGROUND: The use of cyclosporin (CsA) has improved graft survival in transplant (Tx) patients despite its potential nephrotoxicity. Children born to transplanted women may present with intrauterine growth retardation (IUGR). On the basis of potential reduced nephron mass both in IUGR and in newborn experimental animals exposed to CsA in utero, we investigated the renal function of children >1 year of age born to women under maintenance immunosuppression, including CsA. METHODS: Fourteen children born to 12 Tx women (nine kidney, one pancreas-kidney, one heart, one liver) were investigated using inulin clearance (C(in)), para-aminohippuric acid clearance (C(PAH)), microalbuminuria, and electrolyte reabsorption rate. RESULTS: Gestational age of the 14 infants was 34+/-3 weeks and birth weight 2018+/-620 g. During pregnancy, CsA trough blood level was 234+/-115 microg/l and plasma creatinine range was 96-136 micromol/l. Two children were excluded from the study because renal investigation led to a diagnosis of hereditary nephritis (one Alport syndrome, one familial dominant focal segmental glomerulosclerosis) that was retrospectively completed in the mother. Renal function tests were finally performed in 12 children at 2.6+/-1.8 years of age: BP 94+/-7/55+/-5 mmHg, C(in) 117+/-28 ml/min/1.73 m(2), C(PAH) 545+/-124 ml/min/1.73 m(2), filtration fraction 0.23+/-0.03, microalbuminuria 4.2+/-3.5 mg/mmol. Electrolyte tubular reabsorption rates and urine concentrating capacity were normal. CONCLUSION: These results suggest that in children born to transplanted women taking CsA, renal function develops normally despite prolonged exposure in utero. PMID- 11007825 TI - Determinants of urinary excretion of Tamm-Horsfall protein in non-selected kidney stone formers and healthy subjects. AB - BACKGROUND: The aim of the study was to measure urinary excretion of Tamm Horsfall protein (THP), an important inhibitor of crystallization, and to identify possible determinants of urinary THP excretion in non-selected kidney stone formers (SF) and healthy subjects (C). METHODS: By means of a commercially available ELISA (Pharmacia and Upjohn/Elias, Germany), we measured THP in 24-h urines of 104 SF (74 males/30 females, age 16-74 years) who had formed 8.7+/-2.4 stones (range 1-240), and of 71 C (41 males/30 females, age 22-62 years). Types of stones formed by SF were 88 calcium, eight uric acid, six infection, and two cystine. All values are means+/-SE. RESULTS: The normal range (5th to 95th percentile) of U(THP)xV was 9.3-35.0 mg/day in males and 9.0-36.3 mg/day in females respectively. Mean U(THP)xV was 21.3+/-1.2 mg/day (range 3. 4-51.6) in male and 15.2+/-1.6 mg/day (range 1.8-32.3) in female SF (P=0.008 vs male SF). Since U(THP)xV was positively correlated with C(Crea) (r=0.312, P=0.001) in SF as well as with U(Crea)xV (r=0.346, P=0.0001) and with body surface (r=0.271, P=0.0003) in all study subjects, mean THP/Crea (mg/mmol) was used for all further calculations. Overall, THP/Crea was lower in SF (1.42+/-0.07 vs 1. 68+/-0.08, P:=0.015), mainly due to increased THP/Crea in female C (2.08+/-0.11, P=0.0036 vs female SF, P=0.0001 vs male C and vs male calcium SF), which also explains decreased THP/Crea values in calcium SF (1.46+/-0.08, P=0.041 vs C). In addition, THP/Crea was reduced in uric acid SF (1.11+/-0.21, P=0.049 vs C). Whereas THP/Crea was not related to age, urine volume, intake of dairy calcium, or urinary markers of protein intake, either in C or in SF, it correlated significantly with urinary Citrate/Crea, both in C (r=0.523, P=0.0001) and in SF (r=0.221, P=0.025). In C only, but not in SF, THP/Crea was correlated with urinary Calcium/Crea (r=0. 572, P=0.0001) and with Oxalate/Crea (r=0.274, P=0.022). CONCLUSIONS: Both in C and SF, urinary THP excretion is related to body size, renal function and urinary citrate excretion, whereas dietary habits apparently do not affect THP excretion. Uric acid and calcium stone formation predict reduced THP excretion in comparison with C, whereas female gender goes along with increased urinary THP excretion in C. Possibly most relevant to kidney stone formation is the fact that THP excretion rises only in C in response to increasing urinary calcium and oxalate concentrations, whereas this self protective mechanism appears to be missing in SF. PMID- 11007826 TI - Intestinal protein loss in patients with haemorrhagic fever with renal syndrome. AB - BACKGROUND: In haemorrhagic fever with renal syndrome (HFRS) vascular dysfunction has been observed in various organs, but the involvement of the intestine has not yet been reported. This study was performed to evaluate the association of intestinal protein loss in this disease with other clinical parameters reflecting vascular permeability or disease severity. METHODS: Twenty patients with HFRS were included in this study. Intestinal protein loss was measured by (99m)Tc human serum albumin ((99m)Tc-HSA) scintigraphy in the acute stage, and quantitative analysis of protein loss was measured by the faecal clearance of alpha 1-antitrypsin (C(AT)) in the acute and the recovery stages. C(AT) was then compared with clinical parameters reflecting disease activity and vascular permeability. RESULTS: (99m)Tc-HSA scintigraphy was positive in 13 (65%) patients, and C(AT) in the acute stage was significantly increased as compared with C(AT) in the recovery stage (40.5+/-24.1 vs 9.2+/-4.2 ml/day, P<0.001). C(AT) was associated with serum albumin levels, frequency of hypotensive episodes, severity of acute renal failure, and degree of thrombocytopenia. CONCLUSIONS: Our data suggest that the increased vascular permeability of HFRS is associated with the increased intestinal loss of plasma proteins, which might represent one of the parameters of disease severity in HFRS. PMID- 11007827 TI - Incidence and outcome of pauci-immune rapidly progressive glomerulonephritis in Wessex, UK: a 10-year retrospective study. AB - BACKGROUND: The Wessex Renal Unit serves a large stable population (2.5 million). Pauci-immune rapidly progressive glomerulonephritis (RPGN) is a frequent cause of acute renal failure requiring admission to our unit ( approximately 8%). At a population level, little is known of the epidemiology and outcome of RPGN. METHODS: Between 1 April 1986 and 31 March 1996, 141 cases of biopsy proven pauci immune RPGN were seen in the Wessex region. The records of 128 patients were reviewed. Median (range) follow-up was 1.8 (0.9-9.64) years from diagnosis. RESULTS: The incidence of 4 per million was stable throughout the period. No clustering was seen. The diagnosis was made (median, 25th, 75th centile) 78.5 (45, 166) days after symptom onset. Co-morbidity (mostly hypertension) was seen in 47% of patients. Other organs affected were lungs 63%, nose/sinuses 50%, joints 42%, muscle 33%, skin 22% and nervous system 14%. Anti-neutrophil cytoplasmic antibody (ANCA) was positive in 73%; cytoplasmic ANCA 34%, peri nuclear ANCA 26% and undifferentiated 14%. Twenty-seven per cent tested ANCA negative. The differences between the groups were small; time to diagnosis was shorter in the ANCA negative (-ve) group (P=0.02) and there were more airway symptoms in the ANCA positive (+ve) group (P:<0.05). All biopsies demonstrated a necrotizing process; crescents were seen in 96% involving (mean+/-SD) 54+/-26% of the glomeruli. Creatinine concentration (mean+/-SD) at diagnosis was 806+/-540 micromol/l. Treatment followed established immunosuppressive regimens. Initial dialysis was required by 59%, 36% needing long-term dialysis. At 1 year 68% were alive. The need for dialysis (P=0.0004) and age (P=0. 004) were poor prognostic markers. Ten per cent were transplanted, graft survival was 90% at 1 year, no recurrence was seen. CONCLUSIONS: This study, looking at a large cohort, has established the incidence and outcome of ANCA +ve and ANCA -ve RPGN in a defined stable population. It stresses the similarities between ANCA +ve and ANCA -ve cases and supports the notion that pauci-immune RPGN is part of a continuum of vasculitic illness. In this series transplantation is a safe option. PMID- 11007828 TI - Anti-laminin auto antibodies in ANCA-associated vasculitis. AB - BACKGROUND: Endothelial cell damage occurs during vasculitic processes in vivo. With the alteration of the endothelium, exposure to basement membrane components may occur with induction of humoral immunity. METHODS: In the present study, we evaluated the prevalence of antibodies against the basement membrane antigen laminin (LMN) in patients with ANCA-associated systemic vasculitis (AASV), pathologic controls (systemic lupus erythematosus, mixed cryoglobulinaemia, Henoch-Schonlein purpura, primary glomerulonephritis) and normal individuals. RESULTS: By ELISA, 21.6% of AASV (16/74) and 10% of pathologic controls (3/30), but only one of the normal controls (2. 8%) had these antibodies (P=0.02). When AASV patients were divided into two groups according to diagnosis and ANCA antigen specificity, antibodies to LMN were found in 27.5% of MPO-ANCA positive microscopic polyangiitis patients (11/40) vs. only 14.7% of PR3-ANCA positive Wegener granulomatosis patients (5/34). There was no correlation between the presence or titre of anti-LMN antibodies and the main clinical and laboratory parameters. CONCLUSION: These results indicate that basement membrane antigens may become immunogenic in patients with AASV, especially in those with MPO-ANCA positivity. These antibodies are most likely the result of endothelial damage secondary to the initial inflammatory process but may well perpetuate further vascular damage in some patients. PMID- 11007829 TI - Predictors of renal outcome in diffuse proliferative lupus nephropathy: data from repeat renal biopsy. AB - BACKGROUND: Diffuse proliferative lupus nephropathy (DPLN) is the most frequent and severe form of renal disease in patients with systemic lupus erythaematosus. Histological parameters at the initial biopsy of patients with DPLN that would predict the progression of renal pathology or function at the second biopsy are not clearly defined. METHODS: The prognostic significance of renal histological indices, such as glomerular activity index and volume density of cortical interstitium [Vv(int/cortex)], was evaluated from successive renal biopsies in 21 patients with DPLN. RESULTS: At the time of the second biopsies, performed an average of 43 months after the first biopsies, seven patients (33%) showed progressive renal insufficiency. Only three cases (14%) transformed to World Health Organization class I or III. The seven patients with clinical progression exhibited a higher frequency of hypertension, higher percent glomerulosclerosis, and larger Vv(int/cortex) at the time of second biopsy as compared with the 14 patients without renal insufficiency. At the first biopsy, patients with clinical progression showed a higher glomerular activity index (2.9+/-1.2 vs 1.3+/-0.8, P<0.05) and larger Vv(int/cortex) (0.13+/-0.07 microm(3)/microm(3) vs 0.05+/-0.03 microm(3)/microm(3), P<0.05) than the patients without progression. The glomerular activity index at the first biopsy correlated directly with per cent glomerulosclerosis, Vv(int/cortex), and serum creatinine level at the second biopsy. Vv(int/cortex) in the first biopsy also showed a significant relation with per cent glomerulosclerosis and serum creatinine level at the second biopsy. CONCLUSIONS: These results suggest that higher glomerular activity and larger interstitial volume density at the initial biopsy can predict future progression of renal pathology or function in DPLN. PMID- 11007830 TI - Follow-up study of glomerular dimensions and cortical interstitium in microalbuminuric type 1 diabetic patients with or without antihypertensive treatment. AB - BACKGROUND: A decrease in urinary albumin excretion is regularly seen with antihypertensive treatment in patients with diabetic nephropathy. Our study concerns structural data obtained by light microscopy in baseline and follow-up biopsies in antihypertensive treated patients and in a reference group. METHODS: Microalbuminuric type 1 diabetic patients with diabetes duration of 6-16 years were studied. Two groups, allocated to treatment with either angiotensin converting enzyme-inhibitor (group 1, n=6) or beta-blocker (group 2, n=6) after the baseline biopsy, were studied in parallel, whereas the reference group (group 3, n=9), without antihypertensive treatment, was part of a previously completed study. The renal plastic-embedded biopsies were serially sectioned (1 microm), the sections being used for determining glomerular volume, vascular pole area, and interstitial space expressed as fraction of tubular cortex. RESULTS: A significant increase in glomerular volume (P=0.04) was seen in group 3 only. Vascular pole area (VPA) and VPA relative to calculated glomerular surface did not show significant changes in any of the groups, only a tendency to increase in VPA in group 3 (P=0.051). The increase in VPA correlated with systolic blood pressure during the study period (r=0.49, P=0.03). Glomerular volume did not correlate with HbA(1C), current diabetic glomerulopathy, or ensuing worsening of glomerulopathy. In group 3 every case showed an increase in interstitium (P=0.0009), group 2 showed a decrease (P=0.03), and group 1 showed no change. Increase in interstitial fractional volume correlated with diastolic blood pressure during the study (r=0.54, P=0.01). CONCLUSIONS: In early microalbuminuria, type 1 diabetic patients show glomerular growth, probably compensatory to the developing glomerulopathy. The increase in interstitial volume fraction, demonstrable in early nephropathy, is further augmented over a few years, but is arrested by antihypertensive treatment. PMID- 11007831 TI - ACE DD genotype is more susceptible than ACE II and ID genotypes to the antiproteinuric effect of ACE inhibitors in patients with proteinuric non-insulin dependent diabetes mellitus. AB - BACKGROUND: ACE polymorphism, especially genotype DD or D allele, may be involved in the progression of diabetic nephropathy. It may also have different effects on the reduction of proteinuria by ACE inhibitors in patients with proteinuria. We investigated the relationship between ACE gene polymorphism and antiproteinuric effect of ACE inhibitors (Benazepril 10 mg/day or Perindopril 4 mg/day) in 83 NIDDM patients with overt proteinuria (urinary protein excretion over 500 mg/day). METHODS: We recruited NIDDM patients with overt proteinuria from our renal clinic. Before entry, previously used ACE inhibitors were withdrawn for at least 2 weeks and baseline proteinuria and albuminuria were measured. Patients were classified into three groups in accordance with ACE genotypes (17 DD; 33 ID; 33 II) and prospectively followed up for 3 months. Various clinical parameters including age, DM duration, body mass index (BMI), 24-h urine sodium, protein and albumin, BUN, serum creatinine, creatinine clearance (Ccr), mean arterial pressure (MAP), and HbA(1c) were measured in the pre- and post-treatment periods. ACE genotypes were determined by polymerase chain reaction. RESULTS: There were no significant differences in the clinical parameters such as age, DM duration, BMI, BUN, serum creatinine, Ccr, MAP, HbA(1c), and daily urinary excretion of sodium, protein and albumin among three groups (P>0.05). After the 3-month treatment period using ACE inhibitors, there were no significant differences in the reduction of MAP and Ccr among the three groups (P>0.05). However, the percentage reductions in urinary excretion of protein and albumin for DD genotype were significantly higher than in ID and II genotypes (50.9+/-19.2% vs 19.2+/ 16.0%, 20.2+/-20.4%; 52.6+/-23.6% vs. 13.5+/-51.8%, 24.8+/-23.9%, P<0.05). There were no statistically significant correlations between the levels of baseline proteinuria and albuminuria and the magnitudes of the reduction of proteinuria and albuminuria under ACE inhibition (P>0.05). CONCLUSIONS: Our results suggest that the ACE gene polymorphism might have a role in determining the responsiveness to the antiproteinuric effect of ACE inhibition in proteinuric NIDDM patients. PMID- 11007832 TI - Elevated serum levels of soluble adhesion molecules predict death in pre-dialysis patients: association with malnutrition, inflammation, and cardiovascular disease. AB - BACKGROUND: Atherosclerotic cardiovascular disease, malnutrition, and increased levels of pro-inflammatory cytokines are common features in patients with chronic renal failure, and contribute to the high mortality rate observed in these patients. A diverse group of soluble cellular adhesion molecules (CAM) (sVCAM-1, sICAM-1 and sE-selectin) are expressed on the surface of vascular endothelial cells in response to pro-inflammatory cytokines and may play an important role in the atherogenic process. METHODS: Serum levels of sVCAM-1, sICAM-1 (n=87) and sE selectin (n=71) were analysed in a cohort of 88 patients (50+/-1 years) with chronic renal failure. The presence of malnutrition (subjective global assessment (SGA) and serum albumin), inflammation (C-reactive protein (CRP), tumour necrosis factor-alpha (TNF-alpha), and serum hyaluronan), and cardiovascular disease (CVD) were assessed at a time-point close to the start of dialysis treatment (GFR 7+/-1 ml/min). Blood lipid parameters were also assessed. RESULTS: Significant correlations were observed between Log high-sensitivity CRP (hsCRP) and sVCAM-1 (R=0.39; P<0.01) and sICAM-1 (R=0.47; P:<0.001) levels but not between Log hsCRP and sE-selectin levels in 60 patients examined with a hsCRP assay. Also serum concentrations of Log hyaluronan correlated significantly to sVCAM-1 (R=0.34; P<0.01) and sICAM-1 (R=0.29; P<0.05) levels. Malnourished patients (SGA>1) had elevated serum concentrations of sVCAM-1 (1436+/-94 vs. 1105+/-53 ng/ml; P<0.01) compared to well-nourished patients (SGA 1). Patients with clinical signs of CVD (n=26) had elevated serum levels of sICAM-1 (282+/-18 vs. 242+/-9 ng/ml; P<0.05) compared to 61 patients without signs of CVD. Plasma Log lipoprotein (a) (Lp(a)) levels correlated significantly with sVCAM-1 (R=0.30; P<0.01). Survival analysis by the Cox regression model showed that elevated sICAM-1 was, independent of age, SGA, CVD, and Log CRP, significantly related to an increased mortality rate. CONCLUSIONS: Elevated serum concentrations of soluble adhesion molecules are found in pre-dialysis patients who are malnourished, inflamed, and have signs of cardiovascular disease. These data also suggest that sICAM-1 is an independent predictor of mortality in pre-dialysis patients. Further studies are needed to determine if inflammation causes accelerated atherogenesis via effects on soluble adhesion molecules or if elevated serum levels of soluble adhesion molecules are merely markers of endothelial activation in patients with chronic renal failure. PMID- 11007833 TI - Heparins and blood polymorphonuclear stimulation in haemodialysis: an expansion of the biocompatibility concept. AB - BACKGROUND: At the concentrations used in haemodialysis and in a dose-dependent way, unfractionated heparin (UFH) and, to a lesser degree, a low-molecular-weight heparin (LMWH) stimulate polymorphonuclear cells (PMN) in vitro, and could act in synergy with the stimulatory effect of dialysis membranes in vivo. To examine this hypothesis, we studied the effects of different heparin types and regimens on blood PMNs during haemodialysis sessions. METHODS: Ten haemodialysed patients were studied during regular dialysis sessions on a cellulose triacetate membrane (CT 110 G; 1.10 m(2); Baxter), with four different random heparin protocols: one high-UFH regimen (HHR) at 90 IU/kg body-weight (b.w.) and one low-UFH regimen (LHR) at 50 IU/kg b.w., and with a LMWH (nadroparin calcium) at 85 (HHR) or 45 (LHR) IU/kg b.w. Blood granulocytes, platelet counts, and plasma granulocyte degranulation products (elastase, lactoferrin) were measured serially during 4 h dialysis sessions. RESULTS: After 10 min, the reduction in PMNs with UFH was 29.5% for HHR (P<0.01) and 28.5% for LHR (P<0.01), and only 16.8 and 18.6% with LMWH (NS), significantly higher for HHR with UFH than with LMWH (P<0.01). At 60 min, the elastase increase with HHR was greater, 61% with UFH (P<0.01) and 37.8% with LMWH (P<0.01), significantly higher than LHR for UFH (P<0.05) or LMWH (P<0.05). The overall decrease in platelets (with LMWH P<0.01) and the overall increase in lactoferrin (P<0.001) were not different between heparinization procedures. CONCLUSION: Under a conventional heparin regimen, the PMN variation during the course of the dialysis session suggests a more biocompatible effect of LMWH over UFH. In addition, the variation of elastase favours the lower dose, whatever the type of heparin. Heparin type and dose should therefore be considered in studies addressing biocompatibility in haemodialysis: a low dose of LMWH may be viewed as a better biocompatible treatment with regard to leukocyte stimulation. PMID- 11007835 TI - Stewardship or clinical freedom? variations in dialysis decision making. AB - BACKGROUND: It is generally agreed that acceptance criteria for dialysis have varied and changed over time and that implicit rationing, to some extent forced on clinicians by limited capacity, has been widely practised. Our objective was to study the basis and extent of variation in dialysis decision making among nephrologists in one NHS region. DESIGN AND METHODS: In a clinical judgement analysis, linear regression models were employed to reflect the impact of clinical and non-clinical cues on nephrologists' decisions to offer dialysis to 60 'paper patients' under current capacity constraints and under an assumption of no capacity limit. A short questionnaire was also completed by eight nephrologists to elicit their expressed decision drivers, which were subsequently compared with those tacitly derived from the appraisal of the 60 clinical vignettes. RESULTS: Doctors showed substantial variation in their propensity to offer dialysis and in their perceptions of the benefits of dialysis. Even for the five patients where the discordance in propensity to offer dialysis was least, the range in perceived gain in life expectancy was from 24 to 264 months (mean 91 months). The decision models had relatively good explanatory power with an average r(2) of 0.67 (0.39-0.90) and 0.70 (0.47-0.95) for decisions made under current capacity constraints and under an assumption of no limit capacity respectively. Surprisingly, for most doctors, the patient's age had very little impact on dialysis decisions but the magnitude of the beta-coefficients for the patient's mental state (mean -30.7) was of a similar order of magnitude to the coefficient for the principal 'renal' drivers (e.g. the mean coefficient for uraemic symptomatology under current capacity constraints was 47.7). The influence of other non-renal factors on the doctor's likelihood to offer dialysis was largely independent of the capacity assumption. A comparison of the doctor's stated decision drivers with those tacitly derived from their decision models showed only modest correlation. CONCLUSIONS: The extent to which doctors vary in their propensity to offer dialysis is substantial. Very few non-clinical cues appear to influence the decision to offer dialysis. The most important non-renal factor in determining dialysis decisions was the patient's mental state. PMID- 11007834 TI - Role of complement and platelet-activating factor in the stimulation of phagocytosis and reactive oxygen species production during haemodialysis. AB - BACKGROUND: Neutrophil phagocytic functions have been studied extensively in haemodialysis (HD) patients; however, results are contradictory and the mechanisms that modulate phagocytosis and oxidative burst during dialysis are not completely understood. METHODS: The present study investigated neutrophil functions in a selected population of patients before and during clinical dialysis with cuprophane, and polyacrylonitrile (AN69) membranes. We measured phagocytosis of Escherichia coli and intracellular hydrogen peroxide (H2O2) production by flow cytometry in whole blood. RESULTS: Before dialysis, neutrophils from HD patients showed normal phagocytic capability and H2O2 formation. Phagocytosis of FITC-E. coli was significantly stimulated in cuprophane but not AN69-treated patients. Spontaneous and stimulated H2O2 production was enhanced with both cuprophane and AN69 membranes. We then investigated in vitro the role of complement and platelet-activating factor (PAF) in the activation of neutrophils. Incubation of whole blood with C5a increased phagocytosis but not H2O2 production. On the contrary, the addition of synthetic PAF showed a markedly stimulated H2O2 production without increase in phagocytosis. Moreover, during dialysis with formaldehyde-reused cuprophane, complement activation was abolished and phagocytosis was no longer enhanced, while the stimulation of H2O2 production persisted. In addition, we also excluded a particular role of the membrane itself in the activation of neutrophils. CONCLUSION: We demonstrated that in a selected population of HD patients, neutrophils exhibit normal phagocytic capability and normal intracellular H2O2 production. During dialysis, the stimulation of phagocytosis observed with cuprophane is complement dependent, whereas the enhanced H2O2 production observed with both cuprophane and AN69 membranes might be related to PAF production. PMID- 11007836 TI - Impaired glucose oxidation and glucose-induced thermogenesis in renal transplant recipients. AB - BACKGROUND: Renal transplant recipients often show various metabolic abnormalities including reduced glucose tolerance, impaired insulin sensitivity and altered lipid metabolism. However, the acute effects of carbohydrate ingestion on substrate utilization and energy expenditure have not been fully elucidated. METHODS: We evaluated: (i) basal energy expenditure (EE) and substrate utilization, (ii) metabolic fate of an oral glucose load, and (iii) substrate-induced thermogenesis in: (a) 15 non-diabetic renal transplant recipients (Tx) (BMI 25+/-1) on triple immunosuppressive therapy, (b) 11 patients with primary glomerulonephritis (BMI 25+/-1) (Cort) receiving prednisone treatment, and (c) 12 healthy subjects (BMI 26+/-1) (N). Continuous indirect calorimetry was performed in the basal post-absorptive state for 60 min and continued for an additional 180 min following an oral glucose load (75 g). RESULTS: In the basal state, EE was similar in the three study groups. It averaged 14.6+/-0.7, 15.7+/-1.3, and 14.1+/-0.8 cal/kg/min in Tx, Cort, and N respectively. Glucose oxidation was higher in N (1.3+/- 0.2 mg/kg/min) than in Tx (0.7+/-0.2) and Cort (1.0+/-0.2) (P<0.05 in N vs. Tx and vs. Cort), whereas lipid oxidation was lower in N (0.6+/-0.1 mg/kg/min) than in Tx (0.9+/-0.1) and Cort (0.9+/-0.05) (P<0.03 in N vs. Tx and vs. Cort). After glucose ingestion, total carbohydrate oxidation averaged 21.2+/-2, 31.0+/-3, and 29.6+/-3 g, which represented 28+/-3, 41+/-3 and 39+/-2% of the total glucose load in Tx, Cort and N respectively (P<0.01 Tx vs Cort and N). The cumulative increase of EE (180 min) was 9.7+/-2, 13.2+/-3 and 13+/-3 kcal in Tx, Cort, and N respectively. CONCLUSIONS: The present data show that in non-diabetic renal transplant recipients basal EE is normal. However, basal lipid oxidation is higher and glucose oxidation is lower than in healthy subjects. In addition, the oxidative disposal of a glucose load and substrate-induced thermogenesis are impaired. PMID- 11007837 TI - The 825C/T polymorphism of the G-protein subunit beta3 does not influence blood pressure and renal function in kidney transplant recipients. AB - BACKGROUND: Recently, a polymorphism at position 825 (C-->T) of the cDNA that encodes the beta3 subunit of heterotrimeric G proteins (Gbeta3) was found to be associated with essential hypertension. The T allele leads to the formation of a truncated splice variant (Gbeta3-s) with enhanced activity, promoting hypertension. We examined whether the T allele had an influence on blood pressure (BP) and early renal function after renal transplantation. METHODS: We determined the Gbeta3 genotype and T allele frequencies in renal transplant patients and examined associations with BP, BP medications, and renal function in the first year after transplantation. RESULTS: In renal transplant recipients (n=216) the frequency of the T allele was marginally increased (0.34 vs 0.29) compared with normal healthy blood donors (n=163). Age, sex and body mass index were similar in patients with the CC, CT and TT genotype. BP, number of BP medications, and serum creatinine levels were also similar for the three genotypes within the first year after transplantation. Significantly more patients with the TT genotype (48%) had glomerulonephritis as the underlying renal disease, compared with the CT (29%) and CC (27%) genotypes. CONCLUSIONS: The T allele of Gbeta3 does not have a negative impact on BP and early renal function in recipients of a renal allograft. The T allele might play a role in the pathogenesis of chronic glomerulonephritides. PMID- 11007838 TI - Workload generated by a living donor programme for renal transplantation. AB - BACKGROUND: The ethical and medical implications of live kidney donation result in a comprehensive work-up process. The aim of this study was to determine the magnitude of the workload and the yield of renal transplants generated by a live donor programme. METHODS: Referrals to the Leicester live donor programme over the five-year period 1994-1998 were retrospectively assessed. These were initiated by nephrology referral and subsequently investigated in a stepwise manner. Patients were counselled and baseline tests performed prior to consultant surgeon review and assessment of donor renal function/anatomy. RESULTS: One hundred and fifty referrals consisting of 150 recipients with 269 potential donors were originally made. This resulted in 32/120 (27%) related and 3/30 (10%) unrelated recipients (P=0.06) and 32/220 (15%) related and 3/49 (6%) unrelated donors proceeding to live donor transplantation, with a mean work-up time (+/-SD) of 9 (+/-7) months. One hundred and fifteen recipients (77%) and 234 (87%) donors failed to proceed at various stages of assessment, for a variety of immunological, medical and social reasons. A large number of expensive immunological investigations were required for potential donors, the majority of which did not proceed to transplantation. However as a result of performing these in the early stages of assessment the number of more invasive tests is kept to a minimum. CONCLUSIONS: There is a relatively low yield of transplants from live donor referrals, particularly those between unrelated individuals. The vast majority of referrals fail to proceed for legitimate reasons, but as a result, create a significant workload with notable staffing and financial implications. PMID- 11007839 TI - Prevention of acute rejection with antithymocyte globulin, avoiding corticosteroids, and delaying cyclosporin after renal transplantation. AB - BACKGROUND: Despite their well-known side-effects, corticosteroids (Cs) are currently used after kidney transplantation. Avoidance of Cs may improve patient quality of life and eventual long-term survival. We report on a regimen using antithymocyte globulin (ATG) and mycophenolate mofetil (MMF) for induction, and cyclosporin (CsA) plus MMF for maintenance treatment of recipients of primary kidney transplantation. METHODS: We studied 11 consecutive, non-sensitized renal transplant patients (nine cadaver and two living donors). Initial immunosuppression consisted of ATG (1.5 mg/kg/day, i.v.) given for 10 days and MMF (1.0 g/b.i.d.). CsA (8 mg/kg, in two divided doses) was started on post operative day 11. Cs were only allowed in the case of MMF discontinuation, for the treatment of acute rejection, and in the event of recurrence of the primary glomerulonephritis. RESULTS: All patients completed the entire 10-day ATG course. Main side-effects included fever (>38 degrees C) and serum sickness, observed in 73 and 27% of the patients respectively. The incidence of acute rejection was 27% (three of 11 patients). In two patients with acute rejection, serum sickness was concomitantly diagnosed and renal histology was partially compatible with immune complex disease. The remaining patient had two episodes of low-grade rejection. All rejection episodes were rapidly reversed. Two patients (18%) were treated with ganciclovir for cytomegalovirus (CMV) infection. Two patients (18%) are currently receiving Cs for recurrence of the native glomerulonephritis and two rejection episodes respectively. All patients are currently alive with functioning kidneys (average follow-up of 8.4 months; average creatinine level of 128 micromol/l). CONCLUSION: This pilot study suggests that ATG induction in combination with MMF and delayed introduction of CsA, in the absence of Cs, is not well tolerated in recipients of kidney transplants. An earlier introduction of calcineurin inhibitors and/or a shorter course of ATG may reduce the incidence of fever and serum sickness secondary to ATG. PMID- 11007840 TI - Mnemotechnical note on the use of Cockcroft creatinine clearance formula for the validation of a 24-h urine collection. PMID- 11007841 TI - Fibrillary/immunotactoid glomerulonephritis in HIV-positive patients: a report of three cases. PMID- 11007842 TI - Acute tubular necrosis in a patient with Waldenstrom's macroglobulinaemia and hyperviscosity syndrome. PMID- 11007843 TI - Absent pubertal development in a child with chronic renal failure: the case of Frasier syndrome. PMID- 11007844 TI - Tumoral calcinosis associated with pyrexia and systemic inflammatory response in a haemodialysis patient: successful treatment using intravenous pamidronate. PMID- 11007845 TI - The child with haematuria and dysphagia. PMID- 11007847 TI - Cake kidney drained by single ureter: MAG3 renogram for diagnosis and function follow-up. PMID- 11007846 TI - Nephrotic syndrome after renal percutaneous transluminal angioplasty. PMID- 11007848 TI - The simple renal cyst. PMID- 11007849 TI - Unexplained hypertension in a previously normotensive dialysis patient. Diagnosis: pheochromocytoma. PMID- 11007850 TI - Hyponatraemia with natriuresis in neurosurgical patients. PMID- 11007851 TI - Association of Bartter's syndrome with vasculitis. PMID- 11007852 TI - C-reactive protein and cardiovascular disease: linked by complement? PMID- 11007853 TI - Focal sclerosis with tip lesions secondary to polycythaemia vera. PMID- 11007854 TI - Morphology of coronary atherosclerotic lesions in patients with end-stage renal failure. PMID- 11007855 TI - Renal infarction in a patient with von Willebrand disease. PMID- 11007856 TI - Regression of nephrotic syndrome in amyloidosis of familial mediterranean fever following colchicine treatment. PMID- 11007857 TI - Elevating nephrology to a 'stenting' specialty. PMID- 11007858 TI - Successful use of cyclosporin A in progressive anti-glomerular basement membrane nephritis. PMID- 11007859 TI - Haematological toxicity of midodrine in haemodialysis patients. PMID- 11007860 TI - Hepatitis C and holiday dialysis. A postal survey of UK renal units. PMID- 11007861 TI - The utility of AST/ALT ratio as a non-invasive demonstration of the degree of liver fibrosis in chronic HCV patients on long-term haemodialysis. PMID- 11007862 TI - Intravenous ascorbic acid in haemodialysis patients with functional iron deficiency. PMID- 11007863 TI - Changing relationships between serum IL-1, IL-6, and TNF-alpha and dynamic tests of parathyroid gland function in haemodialysis patients with severe hyperparathyroidism in response to calcitriol therapy. PMID- 11007864 TI - Successful treatment of MRSA endocarditis in a haemodialysis patient. PMID- 11007865 TI - Coinfection with leprosy and tuberculosis in a renal transplant recipient. PMID- 11007866 TI - Tacrolimus/itraconazole interactions: a case report of ABO-incompatible living related renal transplantation. PMID- 11007867 TI - Renal transplantation in a patient with Barraquer-Simons disease and mesangiocapillary glomerulonephritis type II. PMID- 11007868 TI - Active hair bundle motion linked to fast transducer adaptation in auditory hair cells. AB - During transduction in auditory hair cells, hair bundle deflection opens mechanotransducer channels that subsequently reclose or adapt to maintained stimuli, a major component of the adaptation occurring on a submillisecond time scale. Using a photodiode imaging technique, we measured hair bundle motion in voltage-clamped turtle hair cells to search for a mechanical correlate of fast adaptation. Excitatory force steps imposed by a flexible glass fiber attached to the bundle caused an initial movement toward the kinocilium, followed by a fast recoil equivalent to bundle stiffening. The recoil had a time course identical to adaptation of the transducer current, and like adaptation, was most prominent for small stimuli, was slowed by reducing extracellular calcium, and varied with hair cell resonant frequency. In free-standing hair bundles, depolarizations positive to 0 mV evoked an outward current attributable to opening of transducer channels, which was accompanied by a sustained bundle deflection toward the kinocilium. Both processes were sensitive to external calcium concentration and were abolished by blocking the transducer channels with dihydrostreptomycin. The similarity in properties of fast adaptation and the associated bundle motion indicates the operation of a rapid calcium-sensitive force generator linked to the gating of the transducer channels. This force generator may permit stimulus amplification during transduction in auditory hair cells. PMID- 11007869 TI - Selective regulation of N-type Ca channels by different combinations of G-protein beta/gamma subunits and RGS proteins. AB - We examined the effects of G-protein beta and gamma subunit heterodimers on human alpha(1B) (N-type) Ca channels expressed in HEK293 cells. All of the known beta subunits (beta1-beta5) produced voltage-dependent inhibition of alpha(1B) Ca channels, depending on the gamma subunit found in the heterodimer. beta1-beta4 subunits inhibited Ca channels when paired with gamma1-gamma3. However, beta5 subunits only produced inhibition when paired with gamma2. In contrast, heterodimers between beta5 subunits and RGS (regulators of G-protein signaling) proteins containing GGL domains did not produce inhibition of Ca channels. However, GGL domain-containing RGS proteins (e.g., RGS6 and RGS11) did block the ability of Gbeta5/gamma2 heterodimers to inhibit Ca channels. Because all of the G-protein beta subunits are found in the nervous system, we conclude that they may all potentially participate in Ca channel inhibition. The interaction of GGL containing RGS proteins with Gbeta5gamma2 suggests a novel way in which Ca channels can be regulated. PMID- 11007870 TI - Pigment epithelium-derived factor supports normal development of photoreceptor neurons and opsin expression after retinal pigment epithelium removal. AB - Dysfunction of the retinal pigment epithelium (RPE), its loss, or separation from the underlying neural retina results in severe photoreceptor degeneration. Pigment epithelium-derived factor (PEDF) is a glycoprotein with reported neuroprotective and differentiation properties that is secreted in abundance by RPE cells. The "pooling" of PEDF within the interphotoreceptor matrix places this molecule in a prime physical location to affect the underlying neural retina. The purpose of this study was to analyze the morphogenetic activity of PEDF in a model of photoreceptor dysmorphogenesis induced by removal of the RPE. Eyes were dissected from embryonic Xenopus laevis, and the RPE was removed before culturing in medium containing PEDF, PEDF plus anti-PEDF antibodies, or medium alone. Control retinas were maintained with an adherent RPE. Light and electron microscopic analysis was used to examine retinal ultrastructure. Opsin was localized immunocytochemically and quantified as an index of outer segment membranous material and photoreceptor protein expression. Removal of the RPE resulted in an aberrant assembly of photoreceptor outer segments, loss of fine subcellular ultrastructure in photoreceptors, and a reduction in opsin protein levels when compared with control retinas. The addition of PEDF prevented the dysmorphic photoreceptor changes induced by RPE removal. In particular, photoreceptor ultrastructure, outer segment membrane assembly, and steady-state levels of opsin were equivalent to control conditions. Anti-PEDF antibodies completely blocked the morphogenetic activity of PEDF. These results indicate that PEDF is able to mimic the supportive role of the RPE on photoreceptors during the final stages of retinal morphogenesis. PMID- 11007871 TI - AMPA receptor current density, not desensitization, predicts selective motoneuron vulnerability. AB - Spinal motoneurons are more susceptible to AMPA receptor-mediated injury than are other spinal neurons, a property that has been implicated in their selective degeneration in amyotrophic lateral sclerosis (ALS). The aim of this study was to determine whether this difference in vulnerability between motoneurons and other spinal neurons can be attributed to a difference in AMPA receptor desensitization and/or to a difference in density of functional AMPA receptors. Spinal motoneurons and dorsal horn neurons were isolated from embryonic rats and cultured on spinal astrocytes. Single-cell RT-PCR quantification of the relative abundance of the flip and flop isoforms of the AMPA receptor subunits, which are known to affect receptor desensitization, did not reveal any difference between the two cell populations. Examination of AMPA receptor desensitization by patch clamp electrophysiological measurements on nucleated and outside-out patches and in the whole-cell mode also yielded similar results for the two cell groups. However, AMPA receptor current density was two- to threefold higher in motoneurons than in dorsal horn neurons, suggesting a higher density of functional AMPA receptors in motoneuron membranes. Pharmacological reduction of AMPA receptor current density in motoneurons to the level found in dorsal horn neurons eliminated selective motoneuron vulnerability to AMPA receptor activation. These results suggest that the greater AMPA receptor current density of spinal motoneurons may be sufficient to account for their selective vulnerability to AMPA receptor agonists in vitro. PMID- 11007872 TI - Muscarinic stimulation of alpha1E Ca channels is selectively blocked by the effector antagonist function of RGS2 and phospholipase C-beta1. AB - Neuronal alpha1E Ca channel subunits are widely expressed in mammalian brain, where they are thought to form R-type Ca channels. Recent studies have demonstrated that R-type channels contribute to neurosecretion and dendritic Ca influx, but little is known concerning their modulation. Here we show that alpha1E channels are strongly stimulated, and only weakly inhibited, through M1 muscarinic acetylcholine receptors. Both forms of channel modulation are mediated by pertussis toxin-insensitive G-proteins. Channel stimulation is blocked by regulator of G-protein signaling 2 (RGS2) or the C-terminal region of phospholipase C-beta1 (PLCbeta1ct), which have been previously shown to function as GTPase-activating proteins for Galphaq. In contrast, RGS2 and PLCbeta1ct do not block inhibition of alpha1E through M1 receptors. Inhibition is prevented, however, by the C-terminal region of beta-adrenergic receptor kinase 1, which sequesters Gbetagamma dimers. Thus, stimulation of alpha1E is mediated by a pertussis toxin-insensitive Galpha subunit (e.g., Galphaq), whereas inhibition is mediated by Gbetagamma. The ability of RGS2 and PLCbeta1ct to selectively block stimulation indicates these proteins functioned primarily as effector antagonists. In support of this interpretation, RGS2 prevented stimulation of alpha1E with non-hydrolyzable guanosine 5'-0-(3-thiotriphosphate). We also report strong muscarinic stimulation of rbE-II, a variant alpha1E Ca channel that is insensitive to voltage-dependent inhibition. Our results predict that Galphaq coupled receptors predominantly stimulate native R-type Ca channels. Receptor mediated enhancement of R-type Ca currents may have important consequences for neurosecretion, dendritic excitability, gene expression, or other neuronal functions. PMID- 11007873 TI - Low-affinity blockade of neuronal N-type Ca channels by the spider toxin omega agatoxin-IVA. AB - The recognition of neuronal Ca channel diversity has led to considerable efforts to identify useful classification criteria. Here, we revisit the pharmacological definition of P- and Q-type Ca channels, which is based on their respective high and low sensitivity to the spider omega-agatoxin-IVA (omega-Aga-IVA), using whole cell recordings of the Ca channel currents carried by 5 mM Ba(2+) in isolated rat subthalamic and sympathetic neurons. In subthalamic neurons, omega-Aga-IVA (1 microM) targeted multiple Ca channels. One population was blocked with high potency. These channels carried 50.4 +/- 3.4% (n = 5) of the control current and showed the same inactivation kinetics and voltage-dependent high affinity for omega-Aga-IVA as do prototypic P-type Ca channels. Other Ca channels were targeted with weaker potency. This heterogeneous population contributed to 14.0 +/- 1.7% (n = 5) of the control current. It included N-type Ca channels as well as high-threshold Ca channels that displayed the pharmacological signature of Q type Ca channels but resembled P-type Ca channels in their gating properties. N type Ca current block by omega-Aga-IVA (1 microM) was further investigated in sympathetic neurons, which mainly express this Ca channel type. Block was incomplete ( approximately 30% of the control current). Its relief at positive potentials was consistent with omega-Aga-IVA acting as a channel-gating modifier. These effects did not reflect a complete loss of selectivity, because omega-Aga IVA (1 microM) had no effect on subthalamic Na and K currents or their T- and L type Ca currents. Our data confirm that omega-Aga-IVA is a selective P-type Ca channel blocker. However, its diminished selectivity in the micromolar range limits its usefulness for functional studies of Q-type Ca channels. PMID- 11007874 TI - Transient NMDA receptor inactivation provides long-term protection to cultured cortical neurons from a variety of death signals. AB - NMDA receptor antagonists, such as (+)-5-methyl-10, 11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine maleate (MK-801), potently block glutamate-induced neuronal death in myriad in vitro cell models and effectively attenuate ischemic damage in vivo. In this report, a novel role for MK-801 and other NMDA receptor antagonists in preconditioning neurons to withstand a wide range of subsequent lethal insults is described. A brief 30 min exposure to 0.1 microM MK-801, applied up to 96 hr before a "lethal" insult, protected primary cortical neurons from a diverse group of neurotoxic agents, including NMDA, beta-amyloid, staurosporine, etoposide, and oxygen-glucose deprivation. This neuroprotective preconditioning by MK-801 arose from transient NMDA receptor inactivation, because the noncompetitive NMDA receptor antagonists memantine and nylindin and the competitive antagonist AP-5 gave similar effects. MK-801 protection was dependent on new protein synthesis during the first 2 hr, but not from 2 to 5 hr, after MK-801 exposure. The MK-801 transient did not alter the ability of NMDA to trigger normally lethal [Ca(2+)](i) influx 48 hr later, but it did block early downstream signaling events coupled to NMDA neurotoxicity, including PKC inactivation and the activation of calpain. Moreover, MK-801 protected neurons from staurosporine-induced apoptosis, although caspase activation in these cells was unimpeded. It is likely that the stress associated with transient inactivation of NMDA receptors triggered a rapid compensatory survival response that provided long-term protection from a spectrum of insults, inducing apoptotic and nonapoptotic death. The possibility that MK-801 preconditioning blocks an event common to seemingly diverse death mechanisms suggests it will be an important tool for obtaining a clearer understanding of the salient molecular events at work in neuronal death and survival pathways. PMID- 11007875 TI - Coordinated gating of TRP-dependent channels in rhabdomeral membranes from Drosophila retinas. AB - Using a newly developed dissociation procedure, we isolated the specialized rhabdomeral membranes from Drosophila retinal photoreceptors. From these membranes, we have recorded spontaneous active currents in excised patch, voltage clamp recordings. We observed rapid opening events that closely resembled those ascribed to one class of light-activated channels, TRP. All activity exhibited Ba(2+) permeability, little voltage dependence, and sensitivity to La(3+) block. Mutational analysis indicated that the spontaneous activity present in these membranes was TRP-dependent. Excised patches from wild-type rhabdomeral membranes exhibited a wide range of conductance amplitudes. In addition, large conductance events exhibited many conductance levels in the open state. Block of activity by La(3+) both developed and recovered in a stepwise manner. Our results indicate that TRP-dependent channels have a small unitary conductance and that many channels can be gated coordinately. PMID- 11007876 TI - Protein phosphatase-mediated regulation of protein kinase C during long-term depression in the adult hippocampus in vivo. AB - The neural substrates of learning and memory are thought to involve use-dependent long-term changes in synaptic function, including long-term depression (LTD) of synaptic strength. One biochemical event hypothesized to contribute to the maintenance and expression of LTD is decreased protein phosphorylation, caused by a decrease in protein kinase activity and/or an increase in protein phosphatase activity. We tested whether the activity of protein kinase C (PKC) decreases after the induction of LTD in area CA1 of the adult hippocampus in vivo, and then investigated the mechanism responsible for the LTD-associated alteration in PKC activity. We found that LTD was associated with a significant decrease in both autonomous and cofactor-dependent PKC activity. The decrease in PKC activity was prevented by NMDA receptor blockade and was not accompanied by a decrease in the level of either PKCalpha, beta, gamma, or zeta. Western blot analysis with phosphospecific antibodies revealed that phosphorylation of Ser-657 on the catalytic domain of PKCalpha (Ser-660 on PKCbetaII) was decreased significantly after the induction of LTD, and that this dephosphorylation was prevented by the protein phosphatase inhibitor okadaic acid. The decrease in autonomous and cofactor-dependent PKC activity likewise was prevented by okadaic acid. These findings suggest that LTD in the adult hippocampus in vivo involves a decrease in PKC activity that is mediated, at least in part, by dephosphorylation of the catalytic domain of PKC by protein phosphatases activated after LTD-inducing stimulation. Our findings are consistent with the idea that protein dephosphorylation contributes to the expression of LTD. PMID- 11007877 TI - Mitochondrial membrane potential and glutamate excitotoxicity in cultured cerebellar granule cells. AB - The relationship between changes in mitochondrial membrane potential (Deltapsi(m)) and the failure of cytoplasmic Ca(2+) homeostasis, delayed Ca(2+)deregulation (DCD), is investigated for cultured rat cerebellar granule cells exposed to glutamate. To interpret the single-cell fluorescence response of cells loaded with tetramethylrhodamine methyl ester (TMRM(+)) or rhodamine-123, we devised and validated a mathematical simulation with well characterized effectors of Deltapsi(m) and plasma membrane potential (Deltapsi(P)). Glutamate usually caused an immediate decrease in Deltapsi(m) of <10 mV, attributable to Ca(2+) accumulation rather than enhanced ATP demand, and these cells continued to generate ATP by oxidative phosphorylation until DCD. Cells for which the mitochondria showed a larger initial depolarization deregulated more rapidly. The mitochondria in a subpopulation of glutamate-exposed cells that failed to extrude Ca(2+) that was released from the matrix after protonophore addition were bioenergetically competent. The onset of DCD during continuous glutamate exposure in the presence or absence of oligomycin was associated with a slowly developing mitochondrial depolarization, but cause and effect could not be established readily. In contrast, the slowly developing mitochondrial depolarization after transient NMDA receptor activation occurs before cytoplasmic free Ca(2+) ([Ca(2+)](c)) has risen to the set point at which mitochondria retain Ca(2+). In the presence of oligomycin no increase in [Ca(2+)](c) occurs during this depolarization. We conclude that transient Ca(2+) loading of mitochondria as a consequence of NMDA receptor activation initiates oxidative damage to both plasma membrane Ca(2+) extrusion pathways and the inhibition of mitochondrial respiration. Depending on experimental conditions, one of these factors becomes rate-limiting and precipitates DCD. PMID- 11007878 TI - Potentiation of a voltage-gated proton current in acidosis-induced swelling of rat microglia. AB - Microglia are equipped with a strong proton (H(+)) extrusion pathway, a voltage gated H(+) channel, probably to compensate for the large amount of H(+) generated during phagocytosis; however, little is known about how this channel is regulated in pathological states. Because neural damage is often associated with intracellular and extracellular acidosis, we examined the regulatory mechanisms of the H(+) current of rat spinal microglia in acidic environments. More than 90% of round/amoeboid microglia expressed the H(+) current, which was characterized by slow activation kinetics, dependencies on both intracellular and extracellular pH, and blockage by Zn(2+). Extracellular lactoacidosis, pH 6.8, induced intracellular acidification and cell swelling. Cell swelling was also induced by intracellular dialysis with acidic pipette solutions, pH 5.5-6.8, at normal extracellular pH 7.3 in the presence of Na(+). The H(+) currents were increased in association with cell swelling as shown by shifts of the half-activation voltage to more negative potentials and by acceleration of the activation kinetics. The acidosis-induced cell swelling and the accompanying potentiation of the H(+) current required nonhydrolytic actions of intracellular ATP and were inhibited by agents affecting actin filaments (phalloidin and cytochalasin D). The H(+) current was also potentiated by swelling caused by hypotonic stress. These findings suggest that the H(+) channel of microglia can be potentiated via cell swelling induced by intracellular acidification. This potentiation might operate as a negative feedback mechanism to protect microglia from cytotoxic acidification and hence acidosis-induced swelling in pathological states of the CNS. PMID- 11007879 TI - Phosphatidylinositol 3-kinase is required for the trophic, but not the survival promoting, actions of NGF on sympathetic neurons. AB - Nerve growth factor (NGF) supports target-dependent survival of sympathetic and other neurons during development; however, the NGF-regulated signaling pathways required for survival are not fully understood. Sympathetic neurons are able to abort acutely the cell death pathway initiated by NGF deprivation at early, as well as late, time points after readdition of NGF. We found that NGF-dependent phosphatidylinositol 3-kinase (PI-3-K) activity inhibited an early cell death event proximal to c-Jun phosphorylation. However, PI-3-K activity was not required for NGF to inhibit the translocation of Bax from the cytoplasm to the mitochondria, nor was it required for NGF to inhibit the subsequent release of mitochondrial cytochrome c, two events required for NGF deprivation-induced apoptosis. MEK/MAPK activity did not account for any of these NGF-dependent events. When subjected to long-term PI-3-K inhibition in the presence of NGF, the majority of sympathetic neurons did not die. Those that did die exhibited significant differences in the characteristics of death caused by PI-3-K inhibition as compared with NGF deprivation. Additionally, PI-3-K inhibition in the presence of NGF did not induce release of mitochondrial cytochrome c, indicating that these neurons were unable to complete the apoptotic program. In contrast to its modest effects on survival, inhibition of PI-3-K induced marked decreases in somal diameter and metabolic function, as measured by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, suggesting that PI-3-K is required for the trophic effects of NGF. Therefore, although PI-3 K is important for the trophic effects of NGF, it is not required for survival. Other, or at least additional, signaling pathways contribute to NGF-mediated survival of sympathetic neurons. PMID- 11007880 TI - Homer proteins regulate coupling of group I metabotropic glutamate receptors to N type calcium and M-type potassium channels. AB - Group I metabotropic glutamate receptors (mGluR1 and 5) couple to intracellular calcium pools by a family of proteins, termed Homer, that cross-link the receptor to inositol trisphosphate receptors. mGluRs also couple to membrane ion channels via G-proteins. The role of Homer proteins in channel modulation was investigated by expressing mGluRs and various forms of Homer in rat superior cervical ganglion (SCG) sympathetic neurons by intranuclear cDNA injection. Expression of cross linking-capable forms of Homer (Homer 1b, 1c, 2, and 3, termed long forms) occluded group I mGluR-mediated N-type calcium and M-type potassium current modulation. This effect was specific for group I mGluRs. mGluR2 (group II) mediated inhibition of N-channels was unaltered. Long forms of Homer decreased modulation of N- and M-type currents but did not selectively block distinct G protein pathways. Short forms of Homer, which cannot self-multimerize (Homer 1a and a Homer 2 C-terminal deletion), did not alter mGluR-ion channel coupling. When coexpressed with long forms of Homer, short forms restored the mGluR1a mediated calcium current modulation in an apparent dose-dependent manner. Homer 2b induced cell surface clusters of mGluR5 in SCG neurons. Conversely, a uniform distribution was observed when mGluR5 was expressed alone or with Homer short forms. These studies indicate that long and short forms of Homer compete for binding to mGluRs and regulate their coupling to ion channels. In vivo, the immediate early Homer 1a is anticipated to enhance ion channel modulation and to disrupt coupling to releasable intracellular calcium pools. Thus, Homer may regulate the magnitude and predominate signaling output of group I mGluRs. PMID- 11007881 TI - Calcineurin-mediated BAD dephosphorylation activates the caspase-3 apoptotic cascade in traumatic spinal cord injury. AB - We report here that activation of the caspase-3 apoptotic cascade in spinal cord injury is regulated, in part, by calcineurin-mediated BAD dephosphorylation. BAD, a proapoptotic member of the bcl-2 gene family, is rapidly dephosphorylated after injury, dissociates from 14-3-3 in the cytosol, and translocates to the mitochondria of neurons where it binds to Bcl-x(L). Pretreatment of animals with FK506, a potent inhibitor of calcineurin activity, or MK801, an NMDA glutamate receptor antagonist, blocked BAD dephosphorylation and abolished activation of the caspase-3 apoptotic cascade. These findings extend previous in vitro observations and are the first to implicate the involvement of glutamate-mediated calcineurin activation and BAD dephosphorylation as upstream, premitochondrial signaling events leading to caspase-3 activation in traumatic spinal cord injury. PMID- 11007882 TI - PICK1 interacts with and regulates PKC phosphorylation of mGLUR7. AB - The G-protein-coupled metabotropic glutamate receptor subtype 7a (mGluR7a) is a member of group III metabotropic glutamate receptors that plays an important role as a presynaptic receptor in regulating transmitter release at glutamatergic synapses. Here we report that the protein interacting with C-kinase (PICK1) binds to the C terminus (ct) of mGluR7a. In the yeast two-hybrid system, the extreme ct of mGluR7a was shown to interact with the PSD-95/Discs large/ZO-1 (PDZ) domain of PICK1. Pull-down assays indicated that PICK1 was retained by a glutathione S transferase fusion of ct-mGluR7a. Furthermore, recombinant and native PICK1/mGluR7a complexes were coimmunoprecipitated from COS-7 cells and rat brain tissue, respectively. Confocal microscopy showed that both PICK1 and mGluR7a displayed synaptic colocalization in cultured hippocampal neurons. PICK1 has previously been shown to bind protein kinase C alpha-subunit (PKCalpha), and mGluR7a is known to be phosphorylated by PKC. We show a relationship between these three proteins using recombinant PICK1, mGluR7, and PKCalpha, where they were co-immunoprecipitated as a complex from COS-7 cells. In addition, PICK1 caused a reduction in PKCalpha-evoked phosphorylation of mGluR7a in in vitro phosphorylation assays. These results suggest a role for PICK1 in modulating PKCalpha-evoked phosphorylation of mGluR7a. PMID- 11007883 TI - Phosphorylation of the AMPA receptor subunit GluR2 differentially regulates its interaction with PDZ domain-containing proteins. AB - PSD-95, DLG, ZO-1 (PDZ) domain-mediated protein interactions have been shown to play important roles in the regulation of glutamate receptor function at excitatory synapses. Recent studies demonstrating the rapid regulation of AMPA receptor function during synaptic plasticity have suggested that AMPA receptor interaction with PDZ domain-containing proteins may be dynamically modulated. Here we show that PKC phosphorylation of the AMPA receptor GluR2 subunit differentially modulates its interaction with the PDZ domain-containing proteins GRIP1 and PICK1. The serine residue [serine-880 (Ser880)] in the GluR2 C-terminal sequence (IESVKI) critical for PDZ domain binding is a substrate of PKC and is phosphorylated in vivo. In vitro binding and coimmunoprecipitation studies show that phosphorylation of serine-880 within the GluR2 PDZ ligand significantly decreases GluR2 binding to GRIP1 but not to PICK1. Immunostaining of cultured hippocampal neurons demonstrates that the Ser880-phosphorylated GluR2 subunits are enriched and colocalized with PICK1 in the dendrites, with very little staining observed at excitatory synapses. Interestingly, PKC activation in neurons increases the Ser880 phosphorylation of GluR2 subunits and recruits PICK1 to excitatory synapses. Moreover, PKC stimulation in neurons results in rapid internalization of surface GluR2 subunits. These results suggest that GluR2 phosphorylation of serine-880 may be important in the regulation of the AMPA receptor internalization during synaptic plasticity. PMID- 11007884 TI - Huntingtin expression stimulates endosomal-lysosomal activity, endosome tubulation, and autophagy. AB - An expansion of polyglutamines in the N terminus of huntingtin causes Huntington's disease (HD) and results in the accrual of mutant protein in the nucleus and cytoplasm of affected neurons. How mutant huntingtin causes neurons to die is unclear, but some recent observations suggest that an autophagic process may occur. We showed previously that huntingtin markedly accumulates in endosomal-lysosomal organelles of affected HD neurons and, when exogenously expressed in clonal striatal neurons, huntingtin appears in cytoplasmic vacuoles causing cells to shrink. Here we show that the huntingtin-enriched cytoplasmic vacuoles formed in vitro internalized the lysosomal enzyme cathepsin D in proportion to the polyglutamine-length in huntingtin. Huntingtin-labeled vacuoles displayed the ultrastructural features of early and late autophagosomes (autolysosomes), had little or no overlap with ubiquitin, proteasome, and heat shock protein 70/heat shock cognate 70 immunoreactivities, and altered the arrangement of Golgi membranes, mitochondria, and nuclear membranes. Neurons with excess cytoplasmic huntingtin also exhibited increased tubulation of endosomal membranes. Exogenously expressed human full-length wild-type and mutant huntingtin codistributed with endogenous mouse huntingtin in soluble and membrane fractions, whereas human N-terminal huntingtin products were found only in membrane fractions that contained lysosomal organelles. We speculate that mutant huntingtin accumulation in HD activates the endosomal-lysosomal system, which contributes to huntingtin proteolysis and to an autophagic process of cell death. PMID- 11007885 TI - Changes in expression of two tetrodotoxin-resistant sodium channels and their currents in dorsal root ganglion neurons after sciatic nerve injury but not rhizotomy. AB - Two TTX-resistant sodium channels, SNS and NaN, are preferentially expressed in c type dorsal root ganglion (DRG) neurons and have been shown recently to have distinct electrophysiological signatures, SNS producing a slowly inactivating and NaN producing a persistent sodium current with a relatively hyperpolarized voltage-dependence. An attenuation of SNS and NaN transcripts has been demonstrated in small DRG neurons after transection of the sciatic nerve. However, it is not known whether changes in the currents associated with SNS and NaN or in the expression of SNS and NaN channel protein occur after axotomy of the peripheral projections of DRG neurons or whether similar changes occur after transection of the central (dorsal root) projections of DRG neurons. Peripheral and central projections of L4/5 DRG neurons in adult rats were axotomized by transection of the sciatic nerve and the L4 and L5 dorsal roots, respectively. DRG neurons were examined using immunocytochemical and patch-clamp methods 9-12 d after sciatic nerve or dorsal root lesion. Levels of SNS and NaN protein in the two types of injuries were paralleled by their respective TTX-resistant currents. There was a significant decrease in SNS and NaN signal intensity in small DRG neurons after peripheral, but not central, axotomy compared with control neurons. Likewise, there was a significant reduction in slowly inactivating and persistent TTX-resistant currents in these neurons after peripheral, but not central, axotomy compared with control neurons. These results indicate that peripheral, but not central, axotomy results in a reduction in expression of functional SNS and NaN channels in c-type DRG neurons and suggest a basis for the altered electrical properties that are observed after peripheral nerve injury. PMID- 11007886 TI - Stimulation-evoked increases in cytosolic [Ca(2+)] in mouse motor nerve terminals are limited by mitochondrial uptake and are temperature-dependent. AB - Increases in cytosolic [Ca(2+)] evoked by trains of action potentials (20-100 Hz) were recorded from mouse and lizard motor nerve terminals filled with a low affinity fluorescent indicator, Oregon Green BAPTA 5N. In mouse terminals at near physiological temperatures (30-38 degrees C), trains of action potentials at 25 100 Hz elicited increases in cytosolic [Ca(2+)] that stabilized at plateau levels that increased with stimulation frequency. Depolarization of mitochondria with carbonylcyanide m-chlorophenylhydrazone (CCCP) or antimycin A1 caused cytosolic [Ca(2+)] to rise to much higher levels during stimulation. Thus, mitochondrial Ca(2+) uptake contributes importantly to limiting the rise of cytosolic [Ca(2+)] during repetitive stimulation. In mouse terminals, the stimulation-induced increase in cytosolic [Ca(2+)] was highly temperature-dependent over the range 18 38 degrees C, with greater increases at lower temperatures. At the lower temperatures, application of CCCP continued to depolarize mitochondria but produced a much smaller increase in the cytosolic [Ca(2+)] transient evoked by repetitive stimulation. This result suggests that the larger amplitude of the stimulation-induced cytosolic [Ca(2+)] transient at lower temperatures was attributable in part to reduced mitochondrial Ca(2+) uptake. In contrast, the stimulation-induced increases in cytosolic [Ca(2+)] measured in lizard motor terminals showed little or no temperature-dependence over the range 18-33 degrees C. PMID- 11007887 TI - Synaptic vesicle transporter expression regulates vesicle phenotype and quantal size. AB - While the transporters that accumulate classical neurotransmitters in synaptic vesicles have been identified, little is known about how their expression regulates synaptic transmission. We have used adenoviral-mediated transfection to increase expression of the brain vesicular monoamine transporter VMAT2 and presynaptic amperometric recordings to characterize the effects on quantal release. In presynaptic axonal varicosities of ventral midbrain neurons in postnatal culture, VMAT2 overexpression in small synaptic vesicles increased both quantal size and frequency, consistent with the recruitment of synaptic vesicles that do not normally release dopamine. This was confirmed using noncatecholaminergic AtT-20 cells, in which VMAT2 expression induced the quantal release of dopamine. The ability to increase quantal size in vesicles that were already competent for dopamine release was shown in PC12 cells, in which VMAT2 expression increased the quantal size but not the number of release events. These results demonstrate that vesicle transporters limit the rate of transmitter accumulation and can alter synaptic strength through two distinct mechanisms. PMID- 11007888 TI - GluR3 autoantibodies destroy neural cells in a complement-dependent manner modulated by complement regulatory proteins. AB - GluR3 autoantibodies have been implicated in the development of Rasmussen's encephalitis, a rare neurodegenerative disease of humans characterized by epilepsy and degeneration of a single cerebral hemisphere. GluR3 autoantibodies are found in some Rasmussen's encephalitis patients, and GluR3 antibodies raised in rabbits destroy cultured cortical cells in a complement-dependent manner. In this study, the cellular targets of anti-GluR3 antisera-mediated cytotoxicity were examined in mixed primary neuronal-glial cultures of rat cortex. Unexpectedly, astrocytes were the principal target of the cytotoxic effects as assessed by immunohistochemistry and lactate dehydrogenase activity; neurons were destroyed to a lesser extent. Astrocyte vulnerability was rescued by transfection with complement regulatory proteins, and neuronal resistance was defeated by impairing complement regulatory protein function. Astrocyte death may occur in Rasmussen's encephalitis, and destruction of this cell type may play a critical role in the progression of this disorder. The present findings suggest complement regulatory protein expression may in part determine the nature and severity of Rasmussen's encephalitis and other complement-dependent nervous system diseases and thus underscore the need for a systematic investigation of the expression of all known complement regulatory proteins in healthy and diseased nervous system tissues. PMID- 11007889 TI - The architectural transcription factor high mobility group I(Y) participates in photoreceptor-specific gene expression. AB - The nonhistone chromosomal proteins high mobility group I(Y) [HMG I(Y)] have been shown to function as architectural transcription factors facilitating enhanceosome formation on a variety of mammalian promoters. Specifically, they have been shown to act as a "molecular glue" mediating protein-protein and protein-DNA contacts within the enhanceosome complex. HMG I(Y) proteins are expressed at high levels in embryonic and transformed cells and have been implicated in transcriptional regulation in these cells. Terminally differentiated cells, however, have been reported to express only minimal, if any, HMG I(Y). In contrast to these observations, we show here that adult mouse retinal photoreceptors, which are terminally differentiated cells, express high levels of these proteins. Using retinoblastoma cells as an approximate model, we further demonstrate in transiently transfected cells that inhibition of HMG I(Y) expression and mutation of HMG I(Y) binding sites significantly reduce rhodopsin promoter activity. DNase I footprint analysis indicates that HMG I protein interacts with a discrete site within the rhodopsin proximal promoter. This site overlaps with the binding site for Crx, a paired-like homeodomain transcription factor that is essential for photoreceptor functioning and that when mutated causes several forms of human photoreceptor degeneration. Both biochemical and functional experiments demonstrate that HMG I(Y) physically associate with Crx and that their interaction with DNA is required for high-level transcription of the rhodopsin gene. These data provide the first demonstration that HMG I(Y) can be important for gene activation in terminally differentiated cells. PMID- 11007890 TI - NGF signals through TrkA to increase clathrin at the plasma membrane and enhance clathrin-mediated membrane trafficking. AB - Neurotrophin (NT) signals may be moved from axon terminals to neuron cell bodies via signaling endosomes-organelles in which NTs continue to be bound to their activated receptors. Suggesting that clathrin-coated membranes serve as one source of signaling endosomes, in earlier studies we showed that nerve growth factor (NGF) treatment increased clathrin at the plasma membrane and resulted in colocalization of clathrin with TrkA, the receptor tyrosine kinase for NGF. Strikingly, however, we also noted that most clathrin puncta at the surface of NGF-treated cells did not colocalize with TrkA, raising the possibility that NGF induces a general increase in clathrin-coated membrane formation. To explore this possibility further, we examined the distribution of clathrin in NGF- and BDNF treated cells. NGF signaling in PC12 cells robustly redistributed the adaptor protein AP2 and the clathrin heavy chain (CHC) to surface membranes. Using confocal and epifluorescence microscopy, as well as biochemical assays, we showed the redistribution of clathrin to be attributable to the activation of TrkA. Significantly, NGF signaled through TrkA to induce an increase in clathrin mediated membrane trafficking, as revealed in the increased endocytosis of transferrin. In that BDNF treatment increased AP2 and clathrin at the surface membranes of hippocampal neurons, these findings may represent a physiologically significant response to NTs. We conclude that NT signaling increases clathrin coated membrane formation and clathrin-mediated membrane trafficking and speculate that this effect contributes to their trophic actions via the increased internalization of receptors and other proteins that are present in clathrin coated membranes. PMID- 11007891 TI - Biogenesis of regulated exocytotic carriers in neuroendocrine cells. AB - Ca(2+)-triggered exocytosis is a hallmark of neurosecretory granules, but the cellular pathway leading to the assembly of these regulated exocytotic carriers is poorly understood. Here we used the pituitary AtT-20 cell line to study the biogenesis of regulated exocytotic carriers involved in peptide hormone secretion. We show that immature secretory granules (ISGs) freshly budded from the trans-Golgi network (TGN) exhibit characteristics of unregulated exocytotic carriers. During a subsequent maturation period they undergo an important switch to become regulated exocytotic carriers. We have identified a novel sorting pathway responsible for this transition. The SNARE proteins, VAMP4 and synaptotagmin IV (Syt IV), enter ISGs initially but are sorted away during maturation. Sorting is achieved by vesicle budding from the ISGs, because it can be inhibited by brefeldin A (BFA). Inhibition of this sorting pathway with BFA arrested the maturing granules in a state that responded poorly to stimuli, suggesting that the transition to regulated exocytotic carriers requires the removal of a putative inhibitor. In support of this, we found that overexpression of Syt IV reduced the stimulus-responsiveness of maturing granules. We conclude that secretory granules undergo a switch from unregulated to regulated secretory carriers during biogenesis. The existence of such a switch may provide a mechanism for cells to modulate their secretory activities under different physiological conditions. PMID- 11007892 TI - The human DIMINUTO/DWARF1 homolog seladin-1 confers resistance to Alzheimer's disease-associated neurodegeneration and oxidative stress. AB - In Alzheimer's disease (AD) brains, selected populations of neurons degenerate heavily, whereas others are frequently spared from degeneration. To address the cellular basis for this selective vulnerability of neurons in distinct brain regions, we compared gene expression between the severely affected inferior temporal lobes and the mostly unaffected fronto-parietal cortices by using an mRNA differential display. We identified seladin-1, a novel gene, which was downregulated in large pyramidal neurons in vulnerable regions in AD but not control brains. Seladin-1 is a human homolog of the DIMINUTO/DWARF1 gene described in plants and Caenorhabditis elegans. Its sequence shares similarities with flavin-adenin-dinucleotide (FAD)-dependent oxidoreductases. In human control brain, seladin-1 was highly expressed in almost all neurons. In PC12 cell clones that were selected for resistance against AD-associated amyloid-beta peptide (Abeta)-induced toxicity, both mRNA and protein levels of seladin-1 were approximately threefold higher as compared with the non-resistant wild-type cells. Functional expression of seladin-1 in human neuroglioma H4 cells resulted in the inhibition of caspase 3 activation after either Abeta-mediated toxicity or oxidative stress and protected the cells from apoptotic cell death. In apoptotic cells, however, endogenous seladin-1 was cleaved to a 40 kDa derivative in a caspase-dependent manner. These results establish that seladin-1 is an important factor for the protection of cells against Abeta toxicity and oxidative stress, and they suggest that seladin-1 may be involved in the regulation of cell survival and death. Decreased expression of seladin-1 in specific neurons may be a cause for selective vulnerability in AD. PMID- 11007893 TI - Mechanisms mediating pituitary adenylate cyclase-activating polypeptide depolarization of rat sympathetic neurons. AB - The direct effects of pituitary adenylate cyclase-activating polypeptides (PACAP) on sympathetic neurons were investigated using rat superior cervical ganglion neurons. Electrophysiological and pharmacological analyses were used to evaluate PACAP modulation of sympathetic neuron membrane potentials and to investigate potential ionic and intracellular signaling mechanisms mediating the responses. More than 90% of the sympathetic neurons were depolarized by the PACAP peptides even when stimulated release was blocked, indicating that the PACAP peptides elicited primary responses in the postganglionic neurons. The response profile was consistent for activation of PACAP-selective PAC(1) receptors: nanomolar concentrations of PACAP27 and PACAP38 were required to stimulate depolarization, whereas vasoactive intestinal peptide failed to evoke any response. Furthermore, depolarizations elicited by PACAP27 were reduced by the PAC(1) receptor antagonist PACAP(6-38). Both sodium influx and inhibition of a potassium current contributed to the peptide-induced depolarizations. Activation of neither pertussis toxin- nor cholera toxin-sensitive G-proteins was required for generation of the depolarizations. cAMP and diacylglycerol production and activation of protein kinase A or protein kinase C also were not requisite for the responses. By contrast, phospholipase C (PLC)-dependent inositol 1,4,5 triphosphate (IP(3)) synthesis was crucial to the PACAP-mediated depolarizations. Although calcium release from IP(3)-sensitive stores was not required for the PACAP-induced responses, inhibition of IP(3) receptors reduced the depolarizations. Thus, among the many signal transduction pathways coupled to the PAC(1) receptor, the PACAP-induced depolarization of sympathetic neurons appears to require activation of PLC and subsequent generation of IP(3). PMID- 11007894 TI - Catecholamines are required for the acquisition of secretory responsiveness by sweat glands. AB - The sympathetic innervation of sweat glands undergoes a developmental change in transmitter phenotype from catecholaminergic to cholinergic. Acetylcholine elicits sweating and is necessary for development and maintenance of secretory responsiveness, the ability of glands to produce sweat after nerve stimulation or agonist administration. To determine whether catecholamines play a role in the development or function of this system, we examined the onset of secretory responsiveness in two transgenic mouse lines, one albino and the other pigmented, that lack tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis. Although both lines lack TH, their catecholamine levels differ because tyrosinase in pigmented mice serves as an alternative source for catecholamine synthesis (Rios et al., 1999). At postnatal day 21 (P21), 28 glands on average are active in interdigital hind footpads of albino TH wild-type mice. In contrast, fewer than one gland is active in albino TH null mice, which lack catecholamines in gland innervation. Treatment of albino TH null mice with DOPA, a catecholamine precursor, from P11 to P21 increases the number of active glands to 14. Pigmented TH null mice, which have faint catecholamine fluorescence in the developing gland innervation, possess 12 active glands at P21, indicating that catecholamines made via tyrosinase, albeit reduced from wild-type levels, support development of responsiveness. Gland formation and the appearance of cholinergic markers occur normally in albino TH null mice, suggesting that catecholamines act directly on gland cells to trigger their final differentiation and to induce responsiveness. Thus, catecholamines, like acetylcholine, are essential for the development of secretory responsiveness. PMID- 11007895 TI - Culture in reduced levels of oxygen promotes clonogenic sympathoadrenal differentiation by isolated neural crest stem cells. AB - Isolated neural crest stem cells (NCSCs) differentiate to autonomic neurons in response to bone morphogenetic protein 2 (BMP2) in clonal cultures, but these neurons do not express sympathoadrenal (SA) lineage markers. Whether this reflects a developmental restriction in NCSCs or simply inappropriate culture conditions was not clear. We tested the growth and differentiation potential of NCSCs at approximately 5% O(2), which more closely approximates physiological oxygen levels. Eighty-three percent of p75(+)P(0-) cells isolated from embryonic day 14.5 sciatic nerve behaved as stem cells under these conditions, suggesting that this is a nearly pure population. Furthermore, addition of BMP2 plus forskolin in decreased oxygen cultures elicited differentiation of thousands of cells expressing tyrosine hydroxylase, dopamine-beta-hydroxylase, and the SA lineage marker SA-1 in nearly all colonies. Such cells also synthesized and released dopamine and norepinephrine. These data demonstrate that isolated mammalian NCSCs uniformly possess SA lineage capacity and further suggest that oxygen levels can influence cell fate. Parallel results indicating that reduced oxygen levels can also promote the survival, proliferation, and catecholaminergic differentiation of CNS stem cells (Studer et al., 2000) suggests that neural stem cells may exhibit a conserved response to reduced oxygen levels. PMID- 11007896 TI - Enhanced proliferation, survival, and dopaminergic differentiation of CNS precursors in lowered oxygen. AB - Standard cell culture systems impose environmental oxygen (O(2)) levels of 20%, whereas actual tissue O(2) levels in both developing and adult brain are an order of magnitude lower. To address whether proliferation and differentiation of CNS precursors in vitro are influenced by the O(2) environment, we analyzed embryonic day 12 rat mesencephalic precursor cells in traditional cultures with 20% O(2) and in lowered O(2) (3 +/- 2%). Proliferation was promoted and apoptosis was reduced when cells were grown in lowered O(2), yielding greater numbers of precursors. The differentiation of precursor cells into neurons with specific neurotransmitter phenotypes was also significantly altered. The percentage of neurons of dopaminergic phenotype increased to 56% in lowered O(2) compared with 18% in 20% O(2). Together, the increases in total cell number and percentage of dopaminergic neurons resulted in a ninefold net increase in dopamine neuron yield. Differential gene expression analysis revealed more abundant messages for FGF8, engrailed-1, and erythropoietin in lowered O(2). Erythropoietin supplementation of 20% O(2) cultures partially mimicked increased dopaminergic differentiation characteristic of CNS precursors cultured in lowered O(2). These data demonstrate increased proliferation, reduced cell death, and enhanced dopamine neuron generation in lowered O(2), making this method an important advance in the ex vivo generation of specific neurons for brain repair. PMID- 11007897 TI - Tumor necrosis factor receptor-associated factor 6 (TRAF6) deficiency results in exencephaly and is required for apoptosis within the developing CNS. AB - Tumor necrosis factor receptor-associated factors (TRAFs) are adaptor proteins important in mediating intracellular signaling. We report here that targeted deletion of traf6 greatly increases the frequency of failure of neural tube closure and exencephaly in traf6 (-/-) mice. The penetrance of this defect is influenced by genetic background. Neural tube fusion requires the coordination of several biological processes, including cell migration invoked by contact dependent signaling, cell proliferation, and programmed cell death (PCD). To gain greater insight into the role of TRAF6 in these processes, neural development and migration within the CNS of traf6 (-/-) mice and controls were assessed through temporal examination of a number of immunohistochemical markers. In addition, relative levels of cellular proliferation and PCD were examined throughout embryonic development using bromodeoxyuridine (BrdU) and in situ terminal deoxynucleotidyl transferase-mediated dUTP biotinylated nick end labeling (TUNEL), respectively. The data suggest that loss of TRAF6 does not significantly alter the level of cellular proliferation or the pattern of neural differentiation per se, but rather regulates the level of PCD within specific regions of the developing CNS. Substantial reductions in TUNEL were observed within the ventral diencephalon and mesencephalon in exencephalic traf6 (-/-) embryos. Our results demonstrate a novel and prominent role for TRAF6 in the regional control of PCD within the developing CNS. PMID- 11007898 TI - L-Type calcium channels mediate calcium oscillations in early postnatal Purkinje neurons. AB - Ca(2+) signaling is important in many fundamental neuronal processes including neurotransmission, synaptic plasticity, neuronal development, and gene expression. In cerebellar Purkinje neurons, Ca(2+) signaling has been studied primarily in the dendritic region where increases in local Ca(2+) have been shown to occur with both synaptic events and spontaneous electrical activity involving P-type voltage-gated Ca(2+) channels (VGCCs), the predominant VGCC expressed by Purkinje neurons. Here we show that Ca(2+) signaling is also a prominent feature of immature Purkinje neurons at developmental stages that precede expression of dendritic structure and involves L-type rather than P-type VGCCs. Immature Purkinje neurons acutely dissociated from postnatal day 4-7 rat pups exhibit spontaneous cytoplasmic Ca(2+) oscillations. The Ca(2+) oscillations require entry of extracellular Ca(2+), are blocked by tetrodotoxin, are communicated to the nucleus, and correlate closely with patterns of endogenously generated spontaneous and evoked electrical activity recorded in the neurons. Immunocytochemistry showed that L-, N-, and P/Q-types of VGCCs are present on the somata of the Purkinje neurons at this age. However, only the L-type VGCC antagonist nimodipine effectively antagonized the Ca(2+) oscillations; inhibitors of P/Q and N-type VGCCs were relatively ineffective. Release of Ca(2+) from intracellular Ca(2+) stores significantly amplified the Ca(2+) signals of external origin. These results show that a somatic signaling pathway that generates intracellular Ca(2+) oscillations and involves L-type VGCCs and intracellular Ca(2+) stores plays a prominent role in the Ca(2+) dynamics of early developing Purkinje neurons and may play an important role in communicating developmental cues to the nucleus. PMID- 11007900 TI - Activity-dependent release of endogenous brain-derived neurotrophic factor from primary sensory neurons detected by ELISA in situ. AB - To define activity-dependent release of endogenous brain-derived neurotrophic factor (BDNF), we developed an in vitro model using primary sensory neurons and a modified ELISA, termed ELISA in situ. Dissociate cultures of nodose-petrosal ganglion cells from newborn rats were grown in wells precoated with anti-BDNF antibody to capture released BDNF, which was subsequently detected using conventional ELISA. Conventional ELISA alone was unable to detect any increase in BDNF concentration above control values following chronic depolarization with 40 mM KCl for 72 hr. However, ELISA in situ demonstrated a highly significant increase in BDNF release, from 65 pg/ml in control to 228 pg/ml in KCl-treated cultures. The efficacy of the in situ assay appears to be related primarily to rapid capture of released BDNF that prevents BDNF binding to the cultured cells. We therefore used this approach to compare BDNF release from cultures exposed for 30 min to either continuous depolarization with elevated KCl or patterned electrical field stimulation (50 biphasic rectangular pulses of 25 msec, at 20 Hz, every 5 sec). Short-term KCl depolarization was completely ineffective at evoking any detectable release of BDNF, whereas patterned electrical stimulation increased extracellular BDNF levels by 20-fold. In addition, the magnitude of BDNF release was dependent on stimulus pattern, with high-frequency bursts being most effective. These data indicate that the optimal stimulus profile for BDNF release resembles that of other neuroactive peptides. Moreover, our findings demonstrate that BDNF release can encode temporal features of presynaptic neuronal activity. PMID- 11007899 TI - Transplanted neuroblasts differentiate appropriately into projection neurons with correct neurotransmitter and receptor phenotype in neocortex undergoing targeted projection neuron degeneration. AB - Reconstruction of complex neocortical and other CNS circuitry may be possible via transplantation of appropriate neural precursors, guided by cellular and molecular controls. Although cellular repopulation and complex circuitry repair may make possible new avenues of treatment for degenerative, developmental, or acquired CNS diseases, functional integration may depend critically on specificity of neuronal synaptic integration and appropriate neurotransmitter/receptor phenotype. The current study investigated neurotransmitter and receptor phenotypes of newly incorporated neurons after transplantation in regions of targeted neuronal degeneration of cortical callosal projection neurons (CPNs). Donor neuroblasts were compared to the population of normal endogenous CPNs in their expression of appropriate neurotransmitters (glutamate, aspartate, and GABA) and receptors (kainate-R, AMPA-R, NMDA-R. and GABA-R), and the time course over which this phenotype developed after transplantation. Transplanted immature neuroblasts from embryonic day 17 (E17) primary somatosensory (S1) cortex migrated to cortical layers undergoing degeneration, differentiated to a mature CPN phenotype, and received synaptic input from other neurons. In addition, 23.1 +/- 13.6% of the donor-derived neurons extended appropriate long-distance callosal projections to the contralateral S1 cortex. The percentage of donor-derived neurons expressing appropriate neurotransmitters and receptors showed a steady increase with time, reaching numbers equivalent to adult endogenous CPNs by 4-16 weeks after transplantation. These results suggest that previously demonstrated changes in gene expression induced by synchronous apoptotic degeneration of adult CPNs create a cellular and molecular environment that is both permissive and instructive for the specific and appropriate maturation of transplanted neuroblasts. These experiments demonstrate, for the first time, that newly repopulating neurons can undergo directed differentiation with high fidelity of their neurotransmitter and receptor phenotype, toward reconstruction of complex CNS circuitry. PMID- 11007901 TI - Effect of dexamethasone treatment on maturational changes in the NMDA receptor in sheep brain. AB - The objective of the present study was to examine the effect of antenatal or postnatal treatment with corticosteroids on the NMDA receptor, one of the mediators of both normal brain development and hypoxic-ischemic injury, by determining the characteristics of the receptor MK-801 binding site in untreated and corticosteroid-treated fetal and newborn lambs. (3)H-MK-801 binding was performed in cerebral cortical cell membranes from fetal sheep at 88, 120, and 136 d gestation (term = 150 d), and from 5-d-old lambs and adult ewes. Animals were randomized to receive dexamethasone [fetuses: 6 mg, i.m. every 12 hr for four doses to mother; lambs: 0.01 mg/kg (low dose) or 0.25 mg/kg (high dose) every 12 hr for four doses] or placebo. During development, B(max) (apparent number of receptors) increased, reaching a maximum in 5-d-old lambs (p < 0.05) and decreasing in the adult brain. K(d) (dissociation constant) did not change, suggesting that receptor affinity was not altered during maturation. Dexamethasone treatment had no effect on MK-801 binding in the fetus or adult, but in lambs was associated with a significant decrease in B(max) from 2.17 +/- 0.18 pmol/mg protein in placebo-treated animals to 1.65 +/- 0.8 and 1.62 +/- 0.07 pmol/mg protein in low-dose and high-dose animals, respectively. Affinity for (3)H-MK-801 decreased 20% after dexamethasone treatment in lambs only (p < 0.05). Thus, dexamethasone treatment appears to modify the NMDA receptor only during a specific period of brain development. PMID- 11007902 TI - Severe hypomyelination of the murine CNS in the absence of myelin-associated glycoprotein and fyn tyrosine kinase. AB - The analysis of mice deficient in the myelin-associated glycoprotein (MAG) or Fyn, a nonreceptor-type tyrosine kinase proposed to act as a signaling molecule downstream of MAG, has revealed that both molecules are involved in the initiation of myelination. To obtain more insights into the role of the MAG-Fyn signaling pathway during initiation of myelination and formation of morphologically intact myelin sheaths, we have analyzed optic nerves of MAG-, Fyn and MAG/Fyn-deficient mice. We observed a slight hypomyelination in optic nerves of MAG mutants that was significantly increased in Fyn mutants and massive in MAG/Fyn double mutants. The severe morphological phenotype of MAG/Fyn mutants, accompanied by behavioral deficits, substantiates the importance of both molecules for the initiation of myelination. The different severity of the phenotype of different genotypes indicates that the MAG-Fyn signaling pathway is complex and suggests the presence of compensatory mechanisms in the single mutants. However, data are also compatible with the possibility that MAG and Fyn act independently to initiate myelination. Hypomyelination of optic nerves was not related to a loss of oligodendrocytes, indicating that the phenotype results from impaired interactions between oligodendrocyte processes and axons and/or impaired morphological maturation of oligodendrocytes. Finally, we demonstrate that Fyn, unlike MAG, is not involved in the formation of ultrastructurally intact myelin sheaths. PMID- 11007903 TI - Expectation of pain enhances responses to nonpainful somatosensory stimulation in the anterior cingulate cortex and parietal operculum/posterior insula: an event related functional magnetic resonance imaging study. AB - Although behavioral studies suggest that pain distress may alter the perception of somatic stimulation, neural correlates underlying such alteration remain to be clarified. The present study was aimed to test the hypothesis that expectation of pain might amplify brain responses to somatosensory stimulation in the anterior cingulate cortex (ACC) and the region including parietal operculum and posterior insula (PO/PI), both of which may play roles in regulating pain-dependent behavior. We compared brain responses with and subjective evaluation of physically identical nonpainful warm stimuli between two psychologically different contexts: one linked with pain expectation by presenting the nonpainful stimuli randomly intermixed with painful stimuli and the other without. By applying the event-related functional magnetic resonance imaging technique, brain responses to the stimuli were assessed with respect to signal changes and activated volume, setting regions of interest on activated clusters in ACC and bilateral PO/PI defined by painful stimuli. As a result, the uncertain expectation of painful stimulus enhanced transient brain responses to nonpainful stimulus in ACC and PO/PI. The enhanced responses were revealed as a higher intensity of signal change in ACC and larger volume of activated voxels in PO/PI. Behavioral measurements demonstrated that expectation of painful stimulus amplified perceived unpleasantness of innocuous stimulus. From these findings, it is suggested that ACC and PO/PI are involved in modulation of affective aspect of sensory perception by the uncertain expectation of painful stimulus. PMID- 11007904 TI - Definition of neuronal circuitry controlling the activity of phrenic and abdominal motoneurons in the ferret using recombinant strains of pseudorabies virus. AB - During a number of behaviors, including vomiting and some postural adjustments, activity of both the diaphragm and abdominal muscles increases. Previous transneuronal tracing studies using injection of pseudorabies virus (PRV) into either the diaphragm or rectus abdominis (RA) of the ferret demonstrated that motoneurons innervating these muscles receive inputs from neurons in circumscribed regions of the spinal cord and brainstem, some of which have an overlapping distribution in the magnocellular part of the medullary reticular formation (MRF). This observation raises two possibilities: that two populations of MRF neurons provide independent inputs to inspiratory and expiratory motoneurons or that single MRF neurons have collateralized projections to both groups of motoneurons. The present study sought to distinguish between these prospects. For this purpose, recombinant isogenic strains of PRV were injected into these respiratory muscles in nine ferrets; the strain injected into the diaphragm expressed beta-galactosidase, whereas that injected into RA expressed green fluorescent protein. Immunofluorescence localization of the unique reporters of each virus revealed three populations of infected premotor neurons, two of which expressed only one virus and a third group that contained both viruses. Dual-infected neurons were predominantly located in the magnocellular part of the MRF, but were absent from both the dorsal and ventral respiratory cell groups. These data suggest that coactivation of inspiratory and expiratory muscles during behaviors such as emesis and some postural adjustments can be elicited through collateralized projections from a single group of brainstem neurons located in the MRF. PMID- 11007905 TI - Parallel streams for the relay of vibrissal information through thalamic barreloids. AB - This study investigated the organization of a vibrissal pathway that arises from the interpolar division of the spinal trigeminal complex (SP5i), transits through the ventral posterior medial nucleus (VPM), and innervates the somatosensory cortical areas in the rat. Using Fluoro-Gold and biotinylated dextran amine, respectively, as retrograde and anterograde tracers, the following organization plan was disclosed. The SP5i projection arises from a population of small-sized neurons that selectively innervate the ventral lateral part of VPM. In cytochrome oxidase-stained material, this region does not display any barreloid arrangement, but Fluoro-Gold injections in single barrel columns labeled rods of cells that extend caudally into the ventral lateral division of VPM. Thus, on the basis of retrograde labeling, barreloids were divided into core and tail compartments, which correspond to the rod segments running across the dorsal and ventral lateral parts of VPM, respectively. Double-labeling experiments revealed that SP5i afferents innervate the tail of barreloids. The anterograde labeling of thalamocortical axons show that most "core cells" project to a single barrel column, whereas some "tail cells" give rise to branching axons that innervate the second somatosensory area and the dysgranular zone of the barrel field. Injections that straddled the transition zone between the core and tail regions disclosed cells projecting to a single barrel column and to the surrounding dysgranular zone. These results suggest that the projection of "barreloids cells" to the granular and/or dysgranular zones relates to the class of prethalamic input(s) they receive. PMID- 11007906 TI - The involvement of recurrent connections in area CA3 in establishing the properties of place fields: a model. AB - Strong constraints on the neural mechanisms underlying the formation of place fields in the rodent hippocampus come from the systematic changes in spatial activity patterns that are consequent on systematic environmental manipulations. We describe an attractor network model of area CA3 in which local, recurrent, excitatory, and inhibitory interactions generate appropriate place cell representations from location- and direction-specific activity in the entorhinal cortex. In the model, familiarity with the environment, as reflected by activity in neuromodulatory systems, influences the efficacy and plasticity of the recurrent and feedforward inputs to CA3. In unfamiliar, novel, environments, mossy fiber inputs impose activity patterns on CA3, and the recurrent collaterals and the perforant path inputs are subject to graded Hebbian plasticity. This sculpts CA3 attractors and associates them with activity patterns in the entorhinal cortex. In familiar environments, place fields are controlled by the way that perforant path inputs select among the attractors. Depending on the training experience provided, the model generates place fields that are either directional or nondirectional and whose changes when the environment undergoes simple geometric transformations are in accordance with experimental data. Representations of multiple environments can be stored and recalled with little interference, and these have the appropriate degrees of similarity in visually similar environments. PMID- 11007907 TI - Cortical feedback controls the frequency and synchrony of oscillations in the visual thalamus. AB - Thalamic circuits have an intrinsic capacity to generate state-dependent oscillations of different frequency and degrees of synchrony, but little is known of how synchronized oscillation is controlled in the intact brain or what function it may serve. The influence of cortical feedback was examined using slice preparations of the visual thalamus and computational models. Cortical feedback was mimicked by stimulating corticothalamic axons, triggered by the activity of relay neurons. This artificially coupled network had the capacity to self-organize and to generate qualitatively different rhythmical activities according to the strength of corticothalamic feedback stimuli. Weak feedback (one to three shocks at 100-150 Hz) phase-locked the spontaneous spindle oscillations (6-10 Hz) in geniculate and perigeniculate nuclei. However, strong feedback (four to eight shocks at 100-150 Hz) led to a more synchronized oscillation, slower in frequency (2-4 Hz) and dependent on GABA(B) receptors. This increase in synchrony was essentially attributable to a redistribution of the timing of action potential generation in lateral geniculate nucleus cells, resulting in an increased output of relay cells toward the cortex. Corticothalamic feedback is thus capable of inducing highly synchronous slow oscillations in physiologically intact thalamic circuits. This modulation may have implications for a better understanding of the descending control of thalamic nuclei by the cortex, and the genesis of pathological rhythmical activity, such as absence seizures. PMID- 11007908 TI - Dissociation in conditioned dopamine release in the nucleus accumbens core and shell in response to cocaine cues and during cocaine-seeking behavior in rats. AB - The dopaminergic innervation of the nucleus accumbens is generally agreed to mediate the primary reinforcing and locomotor effects of psychostimulants, but there is less consensus on conditioned dopamine (DA) release during drug-seeking behavior. We investigated the neurochemical correlates of drug-seeking behavior under the control of a drug-associated cue [a light conditioned stimulus (CS+)] and to noncontingent presentations of the CS+ in the core and shell subregions of the nucleus accumbens. Rats self-administered cocaine under a continuous reinforcement schedule in which a response on one of two identical levers led to an intravenous cocaine infusion (0.25 mg/infusion) and a 20 sec light CS+. Response requirements for cocaine and the CS+ were then progressively increased until stable responding was established under a second-order schedule of reinforcement. During microdialysis, rats were presented noncontingently with a set of 10 sec CS+ and neutral tone stimuli (CS-) before and after a 90 min period during which they responded for cocaine under a second-order schedule. Results showed the following: (1) nucleus accumbens DA increased in both the core and shell during intravenous cocaine self-administration; (2) noncontingent presentations of a cocaine-associated CS+ led to increased DA release selectively in the nucleus accumbens core; and (3) extracellular DA levels were unaltered in both core and shell during a protracted period of drug-seeking behavior under the control of the same cocaine-associated cue. These results indicate that the mesolimbic dopamine system is activated after exposure to drug-associated stimuli under specific conditions. PMID- 11007909 TI - Dissociable roles of mid-dorsolateral prefrontal and anterior inferotemporal cortex in visual working memory. AB - Functional neuroimaging in human subjects and studies of monkeys with lesions limited to the mid-dorsolateral (MDL) prefrontal cortex have shown that this specific region of the prefrontal cortex is involved in visual working memory, although its precise role remains a matter of debate. The present study compared the effect on visual working memory of lesions restricted to the mid-dorsolateral prefrontal cortex of the monkey with that of lesions to the anterior inferotemporal cortex, a region of the temporal cortex specialized for visual memory. Increasing the delay during which information had to be maintained in visual working memory impaired performance after lesions of the anterior inferotemporal cortex, but not after mid-dorsolateral prefrontal lesions. By contrast, increasing the number of stimuli that had to be monitored impaired the performance of animals with mid-dorsolateral prefrontal lesions, but not that of animals with anterior inferotemporal lesions. This demonstration of a double dissociation between the effects of these two lesions provides strong evidence that the role of the mid-dorsolateral prefrontal cortex in visual working memory does not lie in the maintenance of information per se, but rather in the executive process of monitoring this information. In addition, the present study demonstrated that lesions limited to area 9, which constitutes the superior part of the mid-dorsolateral prefrontal region, give rise to a mild impairment in the monitoring of information, whereas lesions of the complete mid-dorsolateral prefrontal region yield a very severe impairment. PMID- 11007914 TI - Interrupting the inhibiton of normal hematopoiesis in myelogenous leukemia: a hypothetical approach to therapy. PMID- 11007915 TI - Suppression of gene expression by targeted disruption of messenger RNA: available options and current strategies. AB - At least three different approaches may be used for gene targeting including: A) gene knockout by homologous recombination; B) employment of synthetic oligonucleotides capable of hybridizing with DNA or RNA, and C) use of polyamides and other natural DNA-bonding molecules called lexitropsins. Targeting mRNA is attractive because mRNA is more accessible than the corresponding gene. Three basic strategies have emerged for this purpose, the most familiar being to introduce antisense nucleic acids into a cell in the hopes that they will form Watson-Crick base pairs with the targeted gene's mRNA. Duplexed mRNA cannot be translated, and almost certainly initiates processes which lead to its destruction. The antisense nucleic acid can take the form of RNA expressed from a vector which has been transfected into the cell, or take the form of a DNA or RNA oligonucleotide which can be introduced into cells through a variety of means. DNA and RNA oligonucleotides can be modified for stability as well as engineered to contain inherent cleaving activity. It has also been proven that because RNA and DNA are very similar chemical compounds, DNA molecules with enzymatic activity could also be developed. This assumption proved correct and led to the development of a "general-purpose" RNA-cleaving DNA enzyme. The attraction of DNAzymes over ribozymes is that they are very inexpensive to make and that because they are composed of DNA and not RNA, they are inherently more stable than ribozymes. Although mRNA targeting is impeccable in theory, many additional considerations must be taken into account in applying these strategies in living cells including mRNA site selection, drug delivery and intracellular localization of the antisense agent. Nevertheless, the ongoing revolution in cell and molecular biology, combined with advances in the emerging disciplines of genomics and informatics, has made the concept of nontoxic, cancer-specific therapies more viable then ever and continues to drive interest in this field. PMID- 11007916 TI - The role of cytokines and adhesion molecules for mobilization of peripheral blood stem cells. AB - CD34(+) hematopoietic stem cells from peripheral blood are commonly used for autologous or allogeneic transplantation following high-dose therapy in malignant diseases. The introduction of hematopoietic growth factors such as G-CSF has greatly facilitated the mobilization of CD34(+) cells. The mechanism of stem cell mobilization is not yet clear. It seems to be a multistep process with a crosstalk between cytokines and adhesion molecules. In this review, the role of hematopoietic growth factors, chemokines, and adhesion molecules for mobilization and homing of CD34(+) cells is summarized. In addition, factors influencing the cytokine-induced mobilization in patients and healthy donors are described. The review closes with an overview of new classes of mobilizing drugs such as monoclonal antibodies, specific peptides, or antisense oligonucleotides targeting adhesion molecules. PMID- 11007917 TI - Thrombopoietin and chemokine mRNA expression in patient post-chemotherapy and in vitro cytokine-treated marrow stromal cell layers. AB - CD34(+) cells and megakaryocyte progenitors were lower in marrow from patients after hematological recovery from the first cycle of 5-fluorouracil, leucovorin, adriamycin, cyclophosphamide (FLAC) chemotherapy plus PIXY321 (GM-CSF/interleukin 3; IL-3 hybrid) than in FLAC + GM-CSF or pre-FLAC marrows. Marrow stromal layers, an in vitro model of the marrow microenvironment, express a combination of stimulatory and inhibitory factors that modulate hematopoietic progenitor cell proliferation and differentiation. The TaqMan assay and quantitative reverse transcriptase-polymerase chain reaction were used to measure monocyte chemoattractant protein-1 (MCP-1), melanoma stimulatory growth activity, and monokine inducible by interferon-gamma (Mig) (inhibitory chemokines for primitive or megakaryocyte progenitors) mRNA levels in in vitro PIXY and GM-CSF-treated and patient post-FLAC marrow stromal layers. Chemokine mRNA was increased after in vitro GM-CSF and to a lesser extent after PIXY treatment. MCP-1 mRNA levels were fivefold higher in FLAC + PIXY than in FLAC + GM-CSF layers, and Mig mRNA was elevated in FLAC + GM-CSF layers. Thrombopoietin (TPO), insulin-like growth factor I (IGF-I), and IGF-II (stimulatory factors for primitive and megakaryocyte progenitors) mRNA were also measured. TPO mRNA levels were 30% lower in GM-CSF and PIXY-pretreated than in control layers with no decrease in IGF mRNA. TPO mRNA in stromal layers of patients who developed grade 3 thrombocytopenia (platelets < 20 x 10(9)/l) during the third cycle of FLAC was only 24% of levels in stromal layers of marrow from other post-FLAC patients. Results demonstrate that patient and in vitro treatment had modulatory effects on TPO and chemokine mRNA expression in marrow stromal layers. PMID- 11007918 TI - New strategies in the treatment of acute myelogenous leukemia: mobilization and transplantation of autologous peripheral blood stem cells in adult patients. AB - During the last decade high-dose Ara-C (HIDAC; single doses of 3 g/m(2)) and autologous stem cell transplantation have been increasingly used as postremission therapy in adult acute myelogenous leukemia (AML). Controlled clinical trials have demonstrated a long-term disease-free survival of 40%-50% for patients treated with at least two courses of HIDAC. Other studies have demonstrated that postremission autologous bone marrow transplantation results in a disease-free survival equal to or better than conventional chemotherapy. However, autotransplantation with mobilized peripheral blood stem cells (PBSC) would now be preferred instead of autologous bone marrow, due to the shorter hematopoietic reconstitution period. The results reviewed in the present article suggest that HIDAC and autologous PBSC transplantation can be combined in the postremission treatment of adult AML, and this combination therapy may also reduce minimal residual disease and the risk of posttransplant relapse. From the available studies it cannot be concluded whether graft purging further reduces the relapse risk. However, the possible advantage of combination therapy with repeated courses of HIDAC and autologous PBSC transplantation needs to be demonstrated in prospective clinical trials before it can be recommended as a part of the routine treatment in AML. PMID- 11007919 TI - Lentiviral vector transduction of hematopoietic stem cells that mediate long-term reconstitution of lethally irradiated mice. AB - Lentiviral vectors efficiently transduce human CD34(+) cells that mediate long term engraftment of nonobese diabetic/severe combined immunodeficient mice. However, hematopoiesis in these animals is abnormal. Typically, 95% of the human cells in peripheral blood are B lymphocytes. To determine whether lentiviral vectors efficiently transduce stem cells that maintain normal hematopoiesis in vivo, we isolated Sca-1(+)c-Kit(+)Lin(-) bone marrow cells from mice without 5 fluorouracil treatment, and transduced these cells in the absence of cytokine stimulation with a novel lentiviral vector containing a GFP (green flourescent protein) reporter gene. These cells were transplanted into lethally irradiated C57Bl/6 mice. In fully reconstituted animals, GFP expression was observed in 8.0% of peripheral blood mononuclear cells for 20 weeks posttransplantation. Lineage analysis demonstrated that a similar percentage (approximately 8.0%) of GFP positive cells was detected in peripheral blood B cells, T cells, granulocytes and monocytes, bone marrow erythroid precursor cells, splenic B cells, and thymic T cells. In secondary transplant recipients, up to 20% of some lineages expressed GFP. Our results suggest that quiescent, hematopoietic stem cells are efficiently transduced by lentiviral vectors without impairing self-renewal and normal lineage specification in vivo. Efficient gene delivery into murine stem cells with lentiviral vectors will allow direct tests of genetic therapies in mouse models of hematopoietic diseases such as sickle cell anemia and thalassemia, in which corrected cells may have a selective survival advantage. PMID- 11007920 TI - The pharmacokinetics and pharmacodynamics of GW395058, a peptide agonist of the thrombopoietin receptor, in the dog, a large-animal model of chemotherapy-induced thrombocytopenia. AB - GW395058, a PEGylated peptide agonist of the thrombopoietin receptor, stimulates megakaryocytopoiesis and has previously been shown to increase platelet counts in vivo. The pharmacokinetics and pharmacodynamics of GW395058 were characterized using a randomized, crossover study in a large-animal model (dog) of chemotherapy induced thrombocytopenia. Nine beagle dogs received i.v. carboplatin (350 mg/m(2)) on day 0 and day 28. GW395058 (1.31 mg/kg) (n = 6) or vehicle control (n = 3) was administered on day 1 and day 29 either as an i.v. bolus or s.c. injection. After i.v. administration, peak concentrations of GW395058 occurred rapidly, while the half-life averaged approximately 56 h. Bioavailability (+/- standard deviation) of GW395058 given s.c. was 78.2% (20.9%). GW395058 (i.v. and s.c.) ameliorated the platelet nadir (p = 0.0086) and resulted in a shorter time to recovery compared to the control group. The mean nadir platelet counts following carboplatin administration were 197,000 cells/microl (80,000) for the i.v. GW395058-dose group, 183,000 cells/microl (72,000) for the s.c.-dose group and 71,000 cells/microl (38,000) for the vehicle-alone group. GW395058 reduced the thrombocytopenic effects of carboplatin in dogs. No GW395058-related adverse side effects were observed. PMID- 11007921 TI - Signaling induced by erythropoietin and stem cell factor in UT-7/Epo cells: transient versus sustained proliferation. AB - UT-7/Epo cells are human factor-dependent erythroleukemic cells, requiring erythropoietin (Epo) for long-term growth. Stem cell factor (SCF) stimulates proliferation of UT-7/Epo only transiently, for three to five days. An investigation of the signal transduction pathways activated by these cytokines in UT-7/Epo cells may identify those signals specifically required for sustained growth. Proliferation assays demonstrate that SCF generates a substantial growth response in UT-7/Epo cells; however, the cells do not multiply or survive past five to seven days. While Epo induces the activation of JAK2 and STAT5, SCF stimulation shows no activation of JAK2 or STATs 1, 3, or 5. The activation of MAPK (p42/44) by SCF was transient, lasting only 30 min, in contrast to Epo, which stimulated phosphorylation of p42/44 for up to 2 h. The expression of the early response genes c-fos, egr1, and cytokine-inducible SH2 protein (CIS) in response to SCF or Epo stimulation demonstrated that the transient expression of p42/44 correlated with the transient expression of c-fos and egr1. In addition, CIS was activated by Epo but not SCF. These data indicate that EpoR, JAK2, and STAT5 activation are not required for the initiation of proliferation of these erythroid cells, that the transient activation of p42/44 correlates with the transient gene expression of c-fos and egr1, and sustained expression of c-fos and egr1 as seen in UT-7/Epo cells continuously grown in Epo may be necessary for long-term proliferation. PMID- 11007922 TI - Characterization of chemokine receptors expressed in primitive blood cells during human hematopoietic ontogeny. AB - Chemokines are capable of regulating a variety of fundamental processes of hematopoietic cells that include proliferation, differentiation, and migration. To evaluate potential chemokine signaling pathways important to the regulation of primitive human hematopoietic cells, we examined chemokine receptor expression of highly purified subpopulations of uncommitted human blood cells. CXCR1-, CXCR2-, CXCR4-, and CCR5-expressing cells were detected by flow cytometry among human blood subsets depleted of lineage-restricted cells (Lin(-)) derived from adult bone marrow, mobilized peripheral blood, cord blood (CB), and circulating fetal blood. Although these chemokine receptors could be detected on Lin(-) cells throughout human development, only CXCR4 could be detected in CD34(-)CD38(-)Lin( ) and CD34(+)CD38(-)Lin(-) subfractions enriched for stem cell function, suggesting that independent of ontogeny, CXCR4-mediated signals are critical to primitive hematopoiesis. Distinct to other stages of human hematopoietic development, primitive CB cells expressed higher levels of CXCR1, CXCR2, CCR5, and CXCR4 on both CD34(-)CD38(-)Lin(-) and CD34(+)CD38(-)Lin(-) subsets. Isolation of these fractions revealed expression of additional chemokine receptors CCR7, CCR8, and Bonzo (STRL133), whereas BOB (GPR15) could not be detected. Our study illustrates that rare uncommitted hematopoietic cells express chemokine receptors not previously associated with primitive human blood cells. Based on these results, we suggest that signaling pathways mediated by chemokine receptors identified here may play a fundamental role in hematopoietic stem cell regulation and provide alternative receptor targets for retroviral pseudotyping for genetic modification of repopulating cells. PMID- 11007923 TI - The molecular perspective: microtubules and the taxanes. PMID- 11007924 TI - Immunotoxin-induced apoptosis. PMID- 11007925 TI - Potential role for NGF in breast cancer. PMID- 11007926 TI - SODD-silencer of death domains. PMID- 11007927 TI - A challenge to solve "Lichtman's riddles". PMID- 11007928 TI - Melanocyte development and malignant melanoma. AB - Malignant melanoma is a notoriously aggressive disease that can affect relatively young individuals and whose incidence is rising at an alarming rate. Unlike many cancers, metastatic melanoma is poorly responsive to current therapies and mutations affecting p53, the retinoblastoma gene product or Ras which occur frequently in many other cancer types, appear to be rare or at least relatively late events in the progression of the disease. Recent advances in our understanding of the disease at the molecular level have indicated that in addition to the loss of cell cycle checkpoints which may be common to all cancers, malignant melanoma shares many characteristics in common with developmental precursors to melanocytes, the mature pigment producing cells of the skin and hair follicles which are responsible for skin and hair colour. This review therefore focuses on the signalling pathways that play a crucial role in the development of the melanocyte lineage which are subject to deregulation in malignant melanoma namely signalling by receptor tyrosine kinases, the Wnt signalling pathway, as well as loss of the p16INK4a cyclin-dependent kinase inhibitor. Intriguingly all three pathways impact on the expression or function of the microphthalmia-associated transcription factor which plays an essential role in melanocyte development. PMID- 11007929 TI - Gene-environment interaction in melanoma. AB - Gene-environment interaction can be defined as a different effect of an environmental exposure in people with different genotypes, or a different effect of a genotype in people with different histories of environmental exposure. Interaction applies when one stratum (high risk) responds differently to an exposure (sun) than another stratum (low risk). Genetic predisposition would appear to be a very important modifier of risk. This paper discusses the concept of gene-environment interaction applied to cutaneous melanoma through discussion of highly penetrant genes and their interaction with sun exposure, through discussion of low penetrant genes and their interaction with sun exposure, and by suggesting a new model for investigation of gene-environment interaction in melanoma. It is stressed that this area of investigation is extremely early in its development. PMID- 11007930 TI - Cytokine-based therapy for melanoma: pre-clinical studies. AB - Incidence and mortality of human malignant melanoma has risen rapidly over recent decades. Systemic therapies for metastatic cutaneous melanoma, the most aggressive of all skin cancers, remain disappointing although immunological treatment has been more successful for melanoma than for most other tumours. With the availability of recombinant cytokines, immunotherapy for melanoma has entered a new era and a growing body of evidence suggests the efficacy of these approaches in pre-clinical models. Cytokine gene transfer to tumour cells has been demonstrated to induce tumour rejection in different murine melanoma models suggesting that vaccination with tumour cells genetically engineered to produce cytokines is an attractive strategy to enhance anti-tumour immune responses in patients with melanoma. Taken together, these data may hold significant promise for the development of effective ex vivo and in vivo systemic and gene therapy modalities to counter the highly metastatic nature of human melanoma. PMID- 11007931 TI - Update on the role of adjuvant interferon for high risk melanoma. AB - Major prognostic factors for melanoma include thickness of the primary lesion, ulceration and presence or absence of regional lymph node metastases. These parameters form the basis for the American Joint Committee on Cancer staging system and the determination of the appropriateness of post-surgical adjuvant therapy. Among the numerous agents tested for the adjuvant therapy of high-risk melanoma, interferon-alpha 2b (IFN-alpha2b) administered at maximally tolerated doses is the only one to demonstrate an improvement in relapse-free and overall survival for these patients. This high-dose IFN-alpha2b regimen comprising an intensive intravenous induction phase followed by a more prolonged subcutaneously administered phase has now been tested in three, large, randomised trials done through the United States Cooperative Groups, and has shown consistent benefit in preventing relapse and improving survival for patients with thick primary melanomas and those with regional lymph node metastases. The relative importance of the induction component of this treatment regimen is being addressed in an ongoing intergroup trial for intermediate-risk melanoma. Data from completed and ongoing studies using high-dose IFN-alpha for the adjuvant therapy of melanoma are presented. PMID- 11007932 TI - Immunologically-mediated tumour cell apoptosis: the role of TRAIL in T cell and cytokine-mediated responses to melanoma. AB - Immune responses against human melanoma are common and are believed to influence the natural history of the disease. In particular, CD4 T cell infiltrates are associated with regression of primary melanoma and with responses to treatment with interferon-alpha2 (IFN-alpha2). Our studies have shown that CD4 T cells appear to kill melanoma by means of a member of the tumour necrosis factor (TNF) family expressed on their surface and called TNF related apoptosis inducing ligand (TRAIL). Moreover, sensitivity to TRAIL also predicts responsiveness of melanoma to CD4 T cells. TRAIL is not expressed on resting lymphocytes but is expressed at high levels after exposure to IFN-alpha2 and on activated T cells. Lymphocytes from melanoma patients in early stages of the disease show high levels of expression after exposure to IFN-alpha2 and IFN-gamma but expression was less on lymphocytes from stage IV patients. This may be due to factors from melanoma cells in that supernatants from some melanoma cultures suppressed IFN alpha2 upregulation of TRAIL. Sensitivity of melanoma cells to TRAIL can be increased by inhibition of the activation of NF-kappaB and anti-apoptotic events downstream of NF-kappaB. These results suggest that TRAIL may be an important mediator of responses against melanoma induced by immunotherapy or by treatment with IFN-alpha2 and interleukin-2. Studies on surgical biopsies of melanoma however show that fresh isolates appear less sensitive to TRAIL-induced apoptosis and effective therapy may involve combinations with other agents. PMID- 11007933 TI - Melanoma antigens recognised by CD8+ and CD4+ T cells. AB - The field of melanoma immunobiology has made tremendous strides in the past decade, resulting in the molecular identification of a vast array of tumour expressed antigens that contain determinants that are recognised by patient T cells or immunoglobulins. The integration of these antigens, their derivative peptides or improved analogues in vaccine trials allows for the augmentation of melanoma-specific CD4+ and CD8+ T cells in situ that may prove clinically efficacious in the adjuvant or therapeutic setting. Indeed, melanoma peptide based immunotherapies targeting the activation of anti-tumour CD8+ cytotoxic T lymphocytes have proven successful (i.e. yielding objective clinical responses), particularly when combined with T cell growth factors or potent antigen presenting cells, such as dendritic cells. Vaccine approaches implementing poly epitope and/or melanoma peptides recognised by CD4+ T cells are anticipated to yield still better clinical outcomes due to the in vivo promotion and maintenance of a diversified, poly-specific effector T cell repertoire directed against resident tumours. PMID- 11007934 TI - New cytotoxic agents for the treatment of metastatic malignant melanoma: temozolomide and related alkylating agents in combination with guanine analogues to abrogate drug resistance. AB - The chloroethylating nitrosoureas (lomustine, fotemustine, cystemustine (BCNU) and methylating agents temozolomide (TMZ), dacarbazine (DTIC), procarbazine) have documented activity in metastatic malignant melanoma with single agent response rates of 15-25%. Chloroethylating agents form chloroethyl adducts at the O6 position of guanine, resulting in N1-guanine, N3-cytosine interstrand crosslinks which are cytotoxic. Methylating agents attack DNA at multiple sites, although most of their cytotoxic activity is due to the formation of methyl adducts at the O6 position of guanine. The presence of these adducts results in a futile recycling of the mismatch repair pathway resulting in DNA strand breaks and apoptotic cell death. An intact mismatch repair system is required to achieve their cytotoxic effect. Repair of adducts by the DNA repair protein O6 alkylguanine DNA alkyltransferase (AGT) impairs the cytotoxic action of both methylating and chloroethylating agents, and mediates a major resistance pathway to these drugs. During DNA repair, irreversible inactivation of AGT occurs. To regenerate AGT activity, synthesis of new molecules is required. Increased but variable AGT activity is found in malignant melanoma, is higher in metastatic lesions than in primary tumours, and is higher in tumours than normal skin. Expression of AGT activity, is higher in melanoma metastases after DTIC chemotherapy compared to expression prior to therapy. TMZ alone depletes human AGT in tumour tissue and peripheral blood progenitor cells. As the t1/2 of TMZ via the oral route is short (approximately 1.8 hours), and the anti-tumour activity of the drug is known to be schedule-dependent, twice daily or prolonged administration schedules of TMZ prevent regeneration of AGT, and render tumour cells more sensitive to the drug. O6-benzylguanine (BG) is a potent AGT inactivating agent. BG and its analogues reduce AGT activity, and increase the in vitro and in vivo efficacy of both methylating and chloroethylating agents. In clinical trials, non-toxic doses of BG deplete AGT to undetectable levels. AGT depleting agents in combination with methylating and chloroethylating agents are now in clinical testing, and may result in greater clinical efficacy in metastatic malignant melanoma. PMID- 11007935 TI - Protein recruitment systems for the analysis of protein-protein interactions. AB - Following the completion of genome projects in a number of organisms, it is becoming evident that a relatively large proportion of the genes identified encode for proteins that have no sequence homology with known proteins. One possible approach towards understanding protein function is to identify the proteins with which a particular protein associates. Although very powerful, the most commonly used genetic method, the two-hybrid system, is limited in its ability to detect all possible protein-protein interactions. The development of novel approaches, such as the protein recruitment systems, provides attractive alternatives towards identification of protein-protein interactions where other methods have failed to function. PMID- 11007936 TI - c-Jun and the transcriptional control of neuronal apoptosis. AB - There has been considerable interest in the molecular mechanisms of apoptosis in mammalian neurons because this form of neuronal cell death is important for the normal development of the nervous system and because inappropriate neuronal apoptosis may contribute to the pathology of human neurodegenerative diseases. The aim of recent research has been to identify the key components of the cell death machinery in neurons and understand how the cell death programme is regulated by intracellular signalling pathways activated by the binding of neurotrophins or death factors to specific cell surface receptors. The aim of this commentary was to review research that has investigated the role of the Jun N-terminal kinase (JNK)/c-Jun signalling pathway in neuronal apoptosis, focusing in particular on work carried out with developing sympathetic neurons. Experiments with sympathetic neurons cultured in vitro, as well as with cerebellar granule neurons and differentiated PC12 cells, have demonstrated that JNK/c-Jun signalling can promote apoptosis following survival factor withdrawal. In addition, experiments with Jnk(-/-) knockout mice have provided evidence that Jnk3 may be required for apoptosis in the hippocampus in vivo following injection of kainic acid, an excitotoxin, and that Jnk1 and Jnk2 are required for apoptosis in the developing embryonic neural tube. However, in the embryonic forebrain, Jnk1 and Jnk2 have the opposite function and are necessary for the survival of developing cortical neurons. These results suggest that JNKs and c-Jun are important regulators of the cell death programme in the mammalian nervous system, but that their biological effects depend on the neuronal type and stage of development. PMID- 11007937 TI - Therapeutic RNA and DNA enzymes. AB - Recent progress in understanding how gene products interact in the control of cell proliferation has engendered high hopes for the rational design of specific therapeutic strategies. The demonstration that certain RNA and DNA nucleic acids can enzymatically cleave mRNAs has offered the possibility of inactivating abnormal gene expression. In principle, this technology is applicable to any disease where a specific gene product can be linked to the initiation and/or perpetuation of the disease. Here, a brief description of the technology that can be useful for the design of therapeutic DNA and RNA agents capable of inducing apoptosis in cancer cells is presented. Furthermore, such agents can be a valuable tool for probing gene function. PMID- 11007939 TI - Apoptosis and nuclear factor-kappa B: a tale of association and dissociation. AB - It is not clear why on treatment with certain killer cytokines or chemotherapeutic agents, some cells undergo apoptosis while others do not. The delineation of sensitivity/resistance pathways should provide a more specific therapy for cancer and other hyperproliferative diseases. Most cells die either by apoptosis or by necrosis. The biochemical pathway that mediates these two modes of cell death has recently been described. The nuclear factor (NF)-kappa B and the genes regulated by this transcription factor have been shown to play a critical role in induction of resistance to killer agents. Thus, inhibitors of NF kappa B activation have a potential in overcoming resistance to apoptosis induced by various agents. The evidence for and against such a notion is discussed. PMID- 11007938 TI - Regulation of the peroxisomal beta-oxidation-dependent pathway by peroxisome proliferator-activated receptor alpha and kinases. AB - The first PPAR (peroxisome proliferator-activated receptor) was cloned in 1990 by Issemann and Green (Nature 347:645-650). This nuclear receptor was so named since it is activated by peroxisome proliferators including several drugs of the fibrate family, plasticizers, and herbicides. This receptor belongs to the steroid receptor superfamily. After activation by a specific ligand, it binds to a DNA response element, PPRE (peroxisome proliferator response element), which is a DR-1 direct repeat of the consensus sequence TGACCT x TGACCT. This mechanism leads to the transcriptional activation of target genes (Motojima et al., J Biol Chem 273:16710-16714, 1998). After the first discovery, several isoforms were characterized in most of the vertebrates investigated. PPAR alpha, activated by hypolipidemic agents of the fibrate family or by leukotrienes; regulates lipid metabolism as well as the detoxifying enzyme-encoding genes. PPAR beta/delta, which is not very well known yet, appears to be more specifically activated by fatty acids. PPAR gamma (subisoforms 1, 2, 3) is activated by the prostaglandin PGJ2 or by antidiabetic thiazolidinediones (Vamecq and Latruffe, Lancet 354:411 418, 1999). This latter isoform is involved in adipogenesis. The level of PPAR expression is largely dependent on the tissue type. PPAR alpha is mainly expressed in liver and kidney, while PPAR beta/delta is almost constitutively expressed. In contrast, PPAR gamma is largely expressed in white adipose tissue. PPAR is a transcriptional factor that requires other nuclear proteins in order to function, i.e. RXRalpha (9-cis-retinoic acid receptor alpha) in all cases in addition to other regulatory proteins. Peroxisomes are specific organelles for very long-chain and polyunsaturated fatty acid catabolism. From our results and those of others, the inventory of the role of PPAR alpha in the regulation of peroxisomal fatty acid beta-oxidation is presented. In relation to this, we showed that PPAR alpha activates peroxisomal beta-oxidation-encoding genes such as acyl-CoA oxidase, multifunctional protein, and thiolase (Bardot et al., FEBS Lett 360:183-186, 1995). Moreover, rat liver PPAR alpha regulatory activity is dependent on its phosphorylated state (Passilly et al., Biochem Pharmacol 58:1001 1008, 1999). On the other hand, some signal transduction pathways such as protein kinase C are modified by peroxisome proliferators that increase the phosphorylation level of some specific proteins (Passilly et al. Eur J Biochem 230:316-321, 1995). From all these findings, PPAR alpha and kinases appear to play an important role in lipid homeostasis. PMID- 11007940 TI - Regulation of nuclear factor-kappa B, activator protein-1, and glutathione levels by tumor necrosis factor-alpha and dexamethasone in alveolar epithelial cells. AB - The development of an oxidant/antioxidant imbalance in lung inflammation may activate redox-sensitive transcription factors such as nuclear factor-kappa B (NF kappa B) and activator protein-1 (AP-1), which regulate the genes for proinflammatory mediators and protective antioxidant genes. GSH, a ubiquitous tripeptide thiol, is a vital intra- and extracellular protective antioxidant against oxidative stress, which plays a key role in the control of proinflammatory processes in the lungs. The rate-limiting enzyme in GSH synthesis is gamma-glutamylcysteine synthetase (gamma-GCS), which consists of a catalytic heavy and a regulatory light subunit. The promoter regions of the human gamma-GCS subunits contain AP-1, NF-kappa B, and antioxidant response elements and are regulated by oxidants, growth factors, inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha), and anti-inflammatory agent (dexamethasone) in lung cells. TNF-alpha depletes intracellular GSH, concomitant with an increase in oxidised glutathione levels in alveolar epithelial cells. TNF-alpha also induces the activation of NF-kappa B and AP-1 and the subsequent increase in gamma-GCS heavy subunit transcription in these cells. Dexamethasone depleted both basal and TNF-alpha-stimulated GSH levels by down-regulating the gamma-GCS-heavy subunit transcription via a mechanism involving AP-1 (c-Jun). The existence of this fine tuning between the redox GSH levels and the activation of transcription factors may determine the balance of transcription for proinflammatory and antioxidant gamma-GCS genes in inflammation. More studies are required to understand the signalling mechanism of the redox regulation of NF-kappa B and AP-1 and gene transcription in inflammation. This could lead to the development of therapeutic strategies based on the pharmacological manipulation of the production of this important antioxidant in inflammation. PMID- 11007941 TI - Cell signaling and regulators of cell cycle as molecular targets for prostate cancer prevention by dietary agents. AB - Prostate cancer (PCA) is the most common invasive malignancy and leading cause (after lung) of cancer deaths in males. Since PCA is initially androgen dependent, strategies are targeted toward androgen depletion for its control. However, tumor re-growth mostly occurs following this modality, and is androgen independent. A loss of functional androgen receptor and an enhanced expression of growth factor receptors (e.g. erbB family members) and associated ligands have been shown to be the causal genetic events in PCA progression. These genetic alterations lead to an epigenetic mechanism where a feed-back autocrine loop between membrane receptor (e.g. epidermal growth factor receptor [erbB1] and associated ligand (e.g. transforming growth factor-alpha) results in an enhanced activation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) as an essential component of the uncontrolled growth of PCA at an advanced and androgen independent stage. Together, we rationalized that inhibiting these epigenetic events would be useful in controlling advanced PCA growth. Dietary polyphenolic flavonoids and isoflavones are being studied extensively as cancer-preventive and interventive agents. Therefore, we focused our attention on silymarin, genistein, and epigallocatechin 3-gallate (EGCG), present in milk thistle, soy beans, and green tea, respectively. The effect of these agents was assessed on the erbB1-Shc ERK1/2 signal transduction pathway, cell cycle regulatory molecules, and cell growth and death. In androgen-independent human prostate carcinoma DU145 cells, silymarin, genistein, and EGCG resulted in a significant to complete inhibition of transforming growth factor-alpha-caused activation of membrane receptor erbB1 followed by inhibition of downstream cytoplasmic signaling target Shc activation and a decrease in its binding with erbB1, without an alteration in their protein expression. Silymarin and genistein also inhibited ERK1/2 activation, suggesting that these agents impair the activation of erbB1-Shc-ERK1/2 signaling in DU145 cells. In the case of EGCG, a further increase in ERK1/2 activation was observed that was related to its pro-oxidant and apoptotic activities. Silymarin, genistein, and EGCG also resulted in a significant induction of Cip1/p21 and Kip1/p27 and a decrease in cyclin-dependent kinase (CDK) 4, but a moderate inhibition of CDK2, cyclin D1, and cyclin E was observed. An enhanced level of Cip1/p21 and Kip1/27 also led to an increase in their binding to CDK4 and CDK2. Treatment of cells with silymarin, genistein, and EGCG also resulted in strong cell growth inhibition at lower doses, and complete inhibition at higher doses. In contrast to silymarin, higher doses of genistein also showed cell death. A more profound cytotoxic effect was observed in the case of EGCG, with strong cell death at lower doses and complete loss of viability at higher doses. Together, these results suggest that cell signaling and regulators of cell cycle are potential epigenetic molecular targets for prostate cancer prevention by dietary agents. More studies, therefore, are needed with these agents to explore their anticarcinogenic potential against human prostate cancer. PMID- 11007942 TI - Inhibitors of prenylation of Ras and other G-proteins and their application as therapeutics. AB - Anchoring of small G-proteins to cellular membranes via a covalently bound lipophylic prenyl group is essential for the functioning of these proteins. For example, the farnesylation of Ras by the action of the enzyme protein:farnesyl transferase (PFT) is pivotal for its signalling function in cell growth and differentiation. The development of inhibitors of PFT was triggered by the role of mutated Ras in certain types of cancer and by the observation that non farnesylated Ras is inactive. Besides the screening of existing compounds for PFT inhibition, rational drug design has also led to new inhibitors. Our research is in the field of atherosclerosis and concerns the development of inhibitors of the growth of vascular smooth muscle cells. The latter process gives rise to reocclusion of the coronary artery (restenosis) after balloon angioplasty. We and others have developed several analogues of the two substrates of PFT, i.e. farnesyl pyrophosphate (FPP) and the so-called CAAX peptide consensus sequence, which were tested in vitro for the inhibition of PFT and of other enzymes involved in protein prenylation, such as protein:geranylgeranyl transferase-1 (PGGT-1). The FPP analogue TR006, a strong inhibitor of PFT (IC(50) of 67 nM), blocked the proliferation of cultured human smooth muscle cells and inhibited platelet-derived growth factor- and basic fibroblast growth factor-induced DNA synthesis. Similar but more highly charged compounds failed in this respect, probably because of an impaired uptake in the cells. Less charged derivatives were designed to circumvent this problem. The effect on the GF-induced activation of intermediates in signal transduction pathways was investigated in order to gain insight into the mechanism of action within the cells. TR006 decreased the bFGF activation of extracellular signal-regulated kinase 1 (ERK1), suggesting its involvement in inhibiting Ras activity. Although other analogues inhibited DNA synthesis, they affected neither ERK1 activation nor p38/stress-activated protein kinase 2 or Jun N-terminal kinase 1 activation. Since some of these compounds were also shown to be inhibitors of in vitro PGGT-1 activity, the geranylgeranylation of other G-proteins may be decreased by these compounds. Rho seems to be a good candidate as a target for inhibitors of PGGT-1. This uncertainty as to the mechanism of action within non-transformed as well as transformed cells applies to all prenylation inhibitors, but is not holding back their further development as drugs. Their current and possible future application as therapeutics in cancer, restenosis, angiogenesis, and osteoporosis is briefly discussed. PMID- 11007943 TI - The integrin alpha IIb/beta 3 in human platelet signal transduction. AB - Platelets are critical for the maintenance of the integrity of the vascular system and are the first line of defence against haemorrhage. When they encounter a subendothelial matrix exposed by injury to a vessel, platelets adhere, are activated, and become adhesive for other platelets so that they aggregate. alpha IIb/beta 3, a platelet-specific integrin, is largely prominent amongst the adhesion receptors and is essential for platelet aggregation. The ligands for alpha IIb/beta 3 are the multivalent adhesive proteins fibrinogen and von Willebrand factor. In resting platelets, alpha IIb/beta 3 is normally in a low activation state, unable to interact with soluble fibrinogen. Stimulation of platelets with various agonists will induce a conformational change in alpha IIb/beta 3 (inside-out signalling), which is then able to bind soluble fibrinogen resulting in the onset of platelet aggregation. However, fibrinogen binding to its membrane receptor is not simply a passive event allowing the formation of intercellular bridges between platelets. Indeed, a complex signalling pathway triggered by integrin ligation and clustering (outside-in signalling) will regulate the extent of irreversible platelet aggregation and clot retraction. Amongst the signalling enzymes activated downstream of alpha IIb/beta 3 engagement, phosphoinositide 3-kinase plays an important role in the control of the irreversible phase of aggregation. PMID- 11007944 TI - Oxidative stress interference with the nuclear factor-kappa B activation pathways. AB - While intracellular redox balance is tightly controlled in many cell types, its modification leads to important cellular changes derived, in part, from a modification of the pattern of gene expression. This modification relies on many transcription factors whose activities are either increased or reduced by a disbalance of the redox environment. Among these transcription factors, nuclear factor-kappa B (NF-kappa B) plays a pivotal role in inducing genes involved in the control of the immune system as well as in the response to injury and infection. Because NF-kappa B can be induced in many cells by a diverse set of stimulating agents, it has been proposed that agents activating it do so by increasing oxidative stress within the cell. However, this model was not found to be universal, since the dependence between NF-kappa B activation and intracellular reactive oxygen species (ROS) generation was only detected in certain cell lines. The origin of this dependency is still unknown, but could very well be situated in a particular kinase or in adaptator molecules of the signaling cascade, leading to inhibitor kappa B alpha (I kappa B alpha phosphorylation. On the other hand, NF-kappa B can be activated by oxidants in many cell types, but this activation is well characterized only in lymphocytes. This activation is distinct from that of classical activators such as proinflammatory cytokines and phorbol esters, because the activation mechanisms appear to converge on a particular tyrosine residue of I kappa B-alpha instead of the two classical N-terminal serines. The nature of the protein kinases or protein phosphatases involved in this process is still undetermined. It will be a challenge in the future to identify the kinases/phosphatases activated by oxidants and to discover why ROS are required in some cells to turn on the transduction pathway leading to NF-kappa B activation by physiological stimuli. PMID- 11007945 TI - Nuclear factor-kappa B, cancer, and apoptosis. AB - The role of nuclear factor (NF)-kappa B in the regulation of apoptosis in normal and cancer cells has been extensively studied in recent years. Constitutive NF kappa B activity in B lymphocytes as well as in Hodgkin's disease and breast cancer cells protects these cells against apoptosis. It has also been reported that NF-kappa B activation by tumor necrosis factor (TNF)-alpha, chemotherapeutic drugs, or ionizing radiations can protect several cell types against apoptosis, suggesting that NF-kappa B could participate in resistance to cancer treatment. These observations were explained by the regulation of antiapoptotic gene expression by NF-kappa B. However, in our experience, inhibition of NF-kappa B activity in several cancer cell lines has a very variable effect on cell mortality, depending on the cell type, the stimulus, and the level of NF-kappa B inhibition. Moreover, in some experimental systems, NF-kappa B activation is required for the onset of apoptosis. Therefore, it is likely that the NF-kappa B antiapoptotic role in response to chemotherapy is cell type- and signal-dependent and that the level of NF-kappa B inhibition is important. These issues will have to be carefully investigated before considering NF-kappa B as a target for genetic or pharmacological anticancer therapies. PMID- 11007946 TI - Induction and regulation of epithelial-mesenchymal transitions. AB - Herein we discuss the factors that bring about the transformation of epithelial cells into cells of fibroblastic phenotype. This type of transformation, referred to as epithelium-to-mesenchyme transition (EMT), allows cells to dissociate from the epithelial tissue from which they originate and to migrate freely. EMT is therefore thought to play a fundamental role during the early steps of invasion and metastasis of carcinoma cells. Among biological agents which have been identified as inducers of EMT are a number of cytokines and extracellular matrix macromolecules. The coordinated changes in cell morphology, associated with the induction of cell motility and the disruption of intercellular junctions, are the consequence of a signaling cascade emanating from the plasma membrane and leading to changes in gene expression. Understanding the mechanisms regulating EMT of normal and transformed epithelial cells may offer new perspectives for designing therapies for the treatment of metastatic cancers of epithelial origin. PMID- 11007947 TI - Inhibition of the type I insulin-like growth factor receptor expression and signaling: novel strategies for antimetastatic therapy. AB - The receptor for the type 1 insulin-like growth factor (IGF-1R) plays a critical role in the acquisition of the malignant phenotype. Using a highly metastatic murine lung carcinoma model, it was demonstrated that this receptor regulates several cellular functions that can impact on the metastatic potential of the cells, including cellular proliferation, anchorage-independent growth, cell migration, and invasion. The tumor model was used to develop several strategies for altering receptor expression and function as means of abrogating the metastatic potential of the cells. They include stable expression in the tumor cells of IGF-1R antisense RNA and dominant negative receptor mutants in which tyrosines in the kinase domain were substituted with phenylalanine. In addition, a novel strategy was used based on altering post ligand-binding receptor turnover. This led to inhibition of receptor re-expression and signaling and resulted in increased tumor cell apoptosis. When combined with the development of viral vectors designed to deliver genetic information with high efficiency, these strategies could form the basis for development of highly specific, antimetastatic therapy in tumors with known IGF-IR involvement. PMID- 11007948 TI - Nuclear factor-kappa B activation and innate immune response in microbial pathogen infection. AB - Human pathogenic microorganisms have developed a variety of strategies to infect the host organism successfully, whereas the host has evolved a series of defense mechanisms. In most cases, the epithelial cell layer represents the first barrier for the bacterial pathogen and triggers the innate and inflammatory responses in the host. Epithelial cells release proinflammatory mediators including cytokines and chemokines, leading to the subsequent attraction of monocytes/macrophages. Therefore, epithelial cells represent an immediate-early warning system in the host organism. Subsequent to the colonization of the epithelial layer, invasive microbial pathogens often induce an acute inflammatory response, which functions to activate residential macrophages and recruits blood leukocytes to the site of infection. Distinct receptors of the Toll family on the cell surface of immune cells mediate antibacterial responses in mammals as well as in Drosophila. One of the most important cellular factors involved in the regulation of the host innate antimicrobial response is the immediate-early response transcription factor nuclear factor (NF)-kappa B. Microbial pathogens activate cellular signal transduction pathways that induce NF-kappa B activation, but pathogens also find ways to overcome the innate immune response through active manipulation of the NF kappa B signal transduction pathways. Exploration of the mechanisms that influence NF-kappa B activity could contribute to a better understanding of the molecular pathogenesis of microbial infections and could be important for potential therapeutic intervention that may be relevant in a wide variety of inflammatory diseases. PMID- 11007949 TI - Integrin-linked kinase (ILK): a "hot" therapeutic target. AB - Integrin-mediated cell adhesion is known to regulate gene expression through the activation of transcription factors. We have recently revealed that these activations are mediated through integrin-linked kinase (ILK). ILK is an ankyrin repeat-containing serine-threonine protein kinase that can interact directly with the cytoplasmic domain of the beta1 and beta3 integrin subunits and whose kinase activity is modulated by cell-extracellular matrix interactions. We have shown that ILK overexpression results in the translocation of beta-catenin to the nucleus, which then forms a complex formation with the lymphoid enhancer binding factor 1 (LEF-1) transcription factor, subsequently activating the transcriptional activity of promoters containing LEF-1 response elements. ILK phosphorylates the glycogen synthase kinase-3 (GSK-3), which inhibits GSK-3 activity. We have demonstrated that ILK stimulates activator protein-1 transcriptional activity through GSK-3 and the subsequent regulation of the c-Jun DNA interaction. ILK also phosphorylates protein kinase B (PKB/Akt) and stimulates its activity. We have shown that ILK is an upstream effector of the phosphatidylinositol 3-kinase-dependent regulation of PKB/Akt. ILK has been shown to phosphorylate PKB/Akt on Ser-473 in vitro and in vivo. Our results clearly indicate that ILK is a key element in the regulation of integrin signaling as well as growth factor and Wnt signaling pathways. PTEN (phosphatase and tensin homolog detected on chromosome 10) is a tumor suppressor gene located on chromosome 10q23 that encodes a protein and phospholipid phosphatase. It is now estimated that inactivation mutants of PTEN exist in 60% of all forms of solid tumors. Loss of expression or mutational inactivation of PTEN leads to the constitutive activation of PKB/Akt via enhanced phosphorylation of Thr-308 and Ser-473. We have demonstrated that the activity of ILK is constitutively elevated in PTEN mutant cells. A small molecule ILK inhibitor suppresses the phosphorylation of PKB at the Ser-473 but not the Thr-308 site in the PTEN mutant cells. These results indicate that inhibition of ILK may be of significant value in solid tumor therapy. PMID- 11007950 TI - Gene regulation of heme oxygenase-1 as a therapeutic target. AB - Heme oxygenase (HO)-1 is the inducible isoform of the rate-limiting enzyme of heme degradation. HO regulates the cellular content of the pro-oxidant heme and produces catabolites with physiological functions. HO-1 is induced by a host of oxidative stress stimuli, and the activation of HO-1 gene expression is considered to be an adaptive cellular response to survive exposure to environmental stresses. Since overexpression of the HO-1 gene is also protective against the deleterious effects of experimental injuries, the specific induction of HO-1 by 'non-stressful' stimuli, eg. stimuli that are not associated with oxidative stress, such as adenosine 3', 5'-cyclic monophosphate or cyclic guanosine 3',5'-monophosphate, may have important clinical implications. This review summarizes recent advances in the understanding of regulatory mechanisms of HO-1 gene expression, in particular the role of various redox-dependent and redox-independent signaling pathways. Models of experimental injuries are highlighted in which specific overexpression of the HO-1 gene either by targeted gene transfer or by pharmacological modulation has been demonstrated to provide therapeutic effects. PMID- 11007952 TI - A20 and A20-binding proteins as cellular inhibitors of nuclear factor-kappa B dependent gene expression and apoptosis. AB - Proper gene expression and cell growth are critical for the survival of all organisms. Nuclear factor-kappa B (NF-kappa B)-dependent gene expression and apoptosis play crucial roles in numerous cellular processes, and defects in their regulation may contribute to a variety of diseases including inflammation and cancer. Although there has recently been tremendous progress in our understanding of the signaling pathways that lead to NF-kappa B activation and apoptosis, signaling mechanisms that negatively regulate these processes are only partially understood. This review deals with the zinc finger protein A20, which has been characterized as a dual inhibitor of NF-kappa B activation and apoptosis. Its inducible expression by a wide variety of stimuli, including cytokines such as tumor necrosis factor, interleukin-1, and CD40, as well as bacterial and viral products such as lipopolysaccharide, Epstein-Barr virus latent membrane protein 1, and human T-cell leukemia virus type I Tax, suggests that it is involved in the negative feedback regulation of signaling. We will discuss the possible underlying mechanisms, placing emphasis on the role of several A20-binding proteins that have recently been described. Moreover, evidence is presented that A20 and A20-binding proteins are potential novel therapeutic tools in the treatment of a variety of diseases. PMID- 11007951 TI - The transcriptional signature of dioxin in human hepatoma HepG2 cells. AB - We have used a high density microarray hybridization approach to characterize the transcriptional response of human hepatoma HepG2 cells to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). We find that exposure to 10 nM TCDD for 8 hr alters by at least a factor of 2.1 the expression of 310 known genes and of an equivalent number of expressed sequence tags. Treatment with TCDD in the presence of 20 microg/mL of cycloheximide blocked the effect on 202 of these genes, allowing us to distinguish between primary effects of TCDD exposure, which take place whether cycloheximide is present or not, and secondary effects, which are blocked by inhibition of protein synthesis. Of the 310 known genes affected by TCDD, 30 are up-regulated and 78 are down-regulated regardless of cycloheximide treatment, and 84 are up-regulated and 118 are down-regulated only when protein synthesis is not inhibited. Functional clustering of genes regulated by TCDD reveals many potential physiological interactions that might shed light on the multiple biological effects of this compound. Our results, however, suggest that arriving at a sound understanding of the molecular mechanisms governing the biological outcome of TCDD exposure promises to be orders of magnitude more complicated than might have been previously imagined. PMID- 11007953 TI - New insights into p53 regulation and gene therapy for cancer. AB - Due to its critical involvement in cell cycle control and apoptotic signaling, the transcription factor p53 has become the most important tumor suppressor currently under investigation. TP53 is the most frequently mutated gene in human cancers and is thought to play a crucial role in malignant transformation. Therefore, p53 appears to be an appealing target for gene therapy. Adenoviral based p53 gene transfection is now being introduced in large clinical trials. Viral cell entry was found to be the rate-limiting step of gene delivery and thus of therapeutic efficiency. Attachment of adenoviruses to the target cell surface is mediated through the coxsackie-adenovirus receptor, and internalization is achieved via interactions with integrins of the alpha v beta(3) and alpha v beta(5) class. The assumption that the restitution of the p53-dependent apoptotic pathway results in a higher responsiveness of solid tumors to cytostatic agents remains a major matter of debate. Combinations of p53-based gene therapy with other components involved in apoptosis, such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/APO2L, or agents neutralizing tumor-promoting antiapoptotic signals, such as humanized anti-growth factor antibodies, should further improve the effectiveness of cancer treatment in the future. PMID- 11007954 TI - Inhibitors of Ras signal transduction as antitumor agents. AB - Anarchic cell proliferation, observed in some leukemia and in breast and ovarian cancers, has been related to dysfunctioning of cytoplasmic or receptor tyrosine kinase activities coupled to p21 Ras. The growth factor receptor-bound protein 2 (Grb2) adaptor when complexed with Sos (Son of sevenless), the exchange factor of Ras, conveys the signal induced by tyrosine kinase-activated receptor to Ras by recruiting Sos to the membrane, allowing activation of Ras. This review shows how it is possible to stop the Ras-deregulated signaling pathway to obtain potential antitumor agents. Grb2 protein is comprised of one SH2 surrounded by two SH3 domains and interacts by means of its Src homology (SH2) domain with phosphotyrosine residues of target proteins such as the epidermal growth factor (EGF) receptor or the Shc adaptor. By means of its SH3 domains, Grb2 recognizes proline-rich sequences of Sos, leading to Ras activation. Inhibitors of SH2 and SH3 domains were designed with the aim of interrupting Grb2 recognition. On the one hand, using structural data and molecular modeling, peptide dimers or "peptidimers", made up of two proline-rich sequences from Sos linked by an optimized spacer, were developed. On the other, using the structure of the Grb2 SH2 domain complexed with a phosphotyrosine (pTyr)-containing peptide and molecular modeling studies, a series of N-protected tripeptides containing two phosphotyrosine or mimetic residues, with one pTyr sterically constrained, were devised. These compounds show very high affinities for Grb2 in vitro. They have been targeted into cells showing selective antiproliferative activity on tumor cells. These results suggest that inhibiting SH2 or SH3 domains of signaling proteins might provide antitumor agents. PMID- 11007955 TI - Signaling angiogenesis via p42/p44 MAP kinase and hypoxia. AB - Angiogenesis is associated with a number of pathological situations. In this study, we have focused our attention on the role of p42/p44 MAP (mitogen activated protein) kinases and hypoxia in the control of angiogenesis. We demonstrate that p42/p44 MAP kinases play a pivotal role in angiogenesis by exerting a determinant action at three levels: i) persistent activation of p42/p44 MAP kinases abrogates apoptosis; ii) p42/p44 MAP kinase activity is critical for controlling proliferation and growth arrest of confluent endothelial cells; and iii) p42/p44 MAP kinases promote VEGF (vascular endothelial growth factor) expression by activating its transcription via recruitment of the AP 2/Sp1 (activator protein-2) complex on the proximal region (-88/-66) of the VEGF promoter and by direct phosphorylation of hypoxia-inducible factor 1 alpha (HIF-1 alpha). HIF-1 alpha plays a crucial role in the control of HIF-1 activity, which mediates hypoxia-induced VEGF expression. We show that oxygen-regulated HIF-1 alpha protein levels are not affected by intracellular localization (nucleus versus cytoplasm). Finally, we propose a model which suggests an autoregulatory feedback mechanism controlling HIF-1 alpha and therefore HIF-1-dependent gene expression. PMID- 11007956 TI - Cyclin A2 transcriptional regulation: modulation of cell cycle control at the G1/S transition by peripheral cues. AB - Several types of cyclins have been identified and among these, cyclin A2 is synthesized in somatic cells at the onset of DNA synthesis as well as during the G2/M transition associated with cyclin-dependent protein kinases 1 and 2. Modulation of cyclin A transcription is due to the interplay between a cell cycle dependent periodic relief of a transcriptional repression and signals transduced through adenosine 3',5'-cyclic monophosphate, transforming growth factor-beta, and the integrin-mediated pathways. Using primary mouse embryonic fibroblasts from embryos where the genes coding for the protein responsible for susceptibility to retinoblastoma (pRB) and the related p107 and p130 proteins had been individually inactivated, we showed that cyclin A is a functional target of pRB-mediated cell cycle arrest. The factors involved are discussed. PMID- 11007957 TI - Signal transduction by tumor necrosis factor and gene regulation of the inflammatory cytokine interleukin-6. AB - Interleukin (IL)-6 is a multifunctional cytokine that can be induced by a plethora of chemical or physiological compounds, including the inflammatory cytokines tumor necrosis factor (TNF) and IL-1. The molecule TNF has a trimeric configuration and thus binds to membrane-bound, cellular receptors to initiate cell death mechanisms and signaling pathways leading to gene induction. Previously, we showed that induced clustering of the intracellular domains of the p55 TNF receptor, or of their respective 'death domains' only, is sufficient to activate the nuclear factor kappa B (NF-kappa B) and several mitogen-activated protein kinase (MAPK) pathways. NF-kappa B is the exclusive transcription factor for induction of the IL-6 gene in response to TNF and functions as the final trigger to activate a multiprotein complex, a so-called 'enhanceosome', at the level of the IL-6 promoter. Furthermore, the enhanceosome displays histone acetylation activity, which turned out to be essential for IL-6 gene activation via NF-kappa B. However, activation of NF-kappa B alone is not sufficient for IL 6 gene induction in response to TNF, as inhibition of the coactivated extracellular signal-regulated kinase and p38 MAPK pathways blocks TNF-mediated gene expression. Nevertheless, the transactivating NF-kappa B subunit p65 is not a direct target of MAPK phosphorylation. Thus, we postulated that other components of the enhanceosome complex are sensitive to MAPK cascades and found that MAPK activity is unequivocally linked to the histone acetylation capacity of the enhanceosome to stimulate gene expression in response to TNF. In contrast, glucocorticoid repression of TNF-driven IL-6 gene expression does not depend on abrogation of histone acetyltransferase activity, but originates from interference of the liganded glucocorticoid receptor with the contacts between NF kappa B p65 and the promoter configuration around the TATA box. PMID- 11007958 TI - The promyelocytic (PML) nuclear compartment and transcription control. AB - Wild-type promyelocytic leukemia (PML) protein and an increasingly documented number of cellular proteins are localized within discrete nuclear structures known as PML nuclear bodies or PODs (potential oncogenic domains). Even though POD function remains elusive, the integrity, topology, and molecular composition of these nuclear compartments have been associated with certain human diseases, including cancer, autoimmunity, neurodegenerative disorders, and viral propagation. At the molecular level, PML protein has been shown to be a coactivator of nuclear hormone receptors, whereas its oncogenic counterpart PML retinoic acid receptor alpha, which promotes POD disaggregation, has been found to activate activator protein-1 transcription in a retinoic acid-dependent manner. Recently, we demonstrated that the CREB-binding protein (CBP) associates with PML protein in vitro and is recruited to the PODs in vivo in a signal dependent manner. In exploring the consequence of this association, we proposed that POD nuclear bodies are regulatory cellular domains where proteins such as the CBP and CBP-interacting molecules may be activated or inactivated to coordinate signal-activated cellular response. This paper discusses the association of PML nuclear bodies with transcription control and underscores the pharmacological aspects of such an observation. PMID- 11007959 TI - Nuclear tyrosine phosphorylation: the beginning of a map. AB - Tyrosine phosphorylation is usually associated with cytoplasmic events. Yet, over the years, many reports have accumulated on tyrosine phosphorylation of individual molecules in the nucleus, and several tyrosine kinases and phosphatases have been found to be at least partially nuclear. The question arises as to whether nuclear tyrosine phosphorylation represents a collection of loose ends of events originating in the cytoplasm or if there may be intranuclear signaling circuits relying on tyrosine phosphorylation to regulate specific processes. The recent discovery of a mechanism causing nuclear tyrosine phosphorylation has prompted us to review the cumulative evidence for nuclear tyrosine phosphorylation pathways and their possible role. While we found that no complex nuclear function has yet been shown to rely upon intranuclear tyrosine phosphorylation in an unambiguous fashion, we found a very high number of compelling observations on individual molecules that suggest underlying networks linking individual events. A systematic proteomics approach to nuclear tyrosine phosphorylation should help chart possible interaction pathways. PMID- 11007960 TI - Role of conformational alteration in the epidermal growth factor receptor (EGFR) function. AB - This mini-review addresses the effect of glycosylation and phosphorylation on the conformational alterations of the epidermal growth factor receptor (EGFR). Based on studies with full-length and truncated EGFRs, we propose a model to suggest that receptor-receptor self-association, which occurs in the truncated receptor and depends on core glycosylation, is prevented in intact receptor by a certain extracellular domain and that the function of the ligand is to remove the negative constraint. We also propose, based on works with a conformation-specific antibody directed to an unphosphorylated peptide, that the interactions among negatively charged phosphotyrosine residues in the receptor molecule result in bringing two epitopes separated by a long stretch of amino acids close to each other to form an antibody-binding site. The implications of these posttranslational modifications on receptor functions are also discussed in this article. PMID- 11007961 TI - Type 2A protein phosphatase, the complex regulator of numerous signaling pathways. AB - Type 2A protein phosphatase (PP2A) comprises a diverse family of phosphoserine- and phosphothreonine-specific enzymes ubiquitously expressed in eukaryotic cells. Common to all forms of PP2A is a catalytic subunit (PP2Ac) which can form two distinct complexes, one with a structural subunit termed PR65/A and another with an alpha4 protein. The PR65/A-PP2Ac dimer may further associate with a regulatory subunit and form a trimeric holoenzyme. To date, three distinct families of regulatory subunits, which control substrate selectivity and phosphatase activity and target PP2A holoenzymes to their substrates, have been identified. Other molecular mechanisms that regulate PP2Ac function include phosphorylation, carboxyl methylation, inhibition by intracellular protein inhibitors (I(1)(PP2A) and I(2)(PP2A)), and stimulation by ceramide. PP2A dephosphorylates many proteins in vitro, but in vivo protein kinases and transcription factors appear to represent two major sets of substrates. Several natural compounds can inhibit PP2A activity and are used to study its function. Mutations in genes encoding PR65/A subunits have been identified in several different human cancers and the PP2A inhibitor, termed fostriecin, is being tested as an anticancer drug. Thus, a more thorough understanding of PP2A structure and function may lead to the development of novel strategies against human diseases. PMID- 11007962 TI - Phosphorylation of the cap-binding protein eIF4E by the MAPK-activated protein kinase Mnk1. AB - The purpose of this review is to summarize recent experimental data describing the regulation of the phosphorylation of eIF4E, the cap-binding protein, by the MAPK-activated protein kinase Mnk1. Mnk1 does not interact directly with eIF4E, but uses a docking site in eIF4G, a partner of eIF4E. Consequently, control of eIF4E phosphorylation may not strictly depend on changes in Mnk1 activity. The possibility that integrity of the eIF4E/eIF4G/Mnk1 complex also impinges upon eIF4E phosphorylation is discussed. PMID- 11007963 TI - Role of the peroxisome proliferator-activated receptors (PPAR) in atherosclerosis. AB - Peroxisome proliferator-activated receptors (PPAR) are ligand-activated transcription factors which form a subfamily of the nuclear receptor gene family. PPAR activators have effects on both metabolic risk factors and on vascular inflammation related to atherosclerosis. PPAR have profound effects on the metabolism of lipoproteins and fatty acids. PPAR alpha binds hypolipidemic fibrates, whereas PPAR gamma has a high affinity for antidiabetic glitazones. Both PPAR are activated by fatty acids and their derivatives. Activation of PPAR alpha increases the catabolism of fatty acids at several levels. In the liver, it increases uptake of fatty acids and activates their beta-oxidation. The effects that PPAR alpha exerts on triglyceride-rich lipoproteins is due to their stimulation of lipoprotein lipase and repression of apolipoprotein CIII expression, while the effects on high-density lipoproteins depend upon the regulation of apolipoproteins AI and AII. PPAR gamma has profound effects on the differentiation and function of adipose tissue, where it is highly expressed. PPAR are also expressed in atherosclerotic lesions. PPAR are present in vascular endothelial cells, smooth muscle cells, monocytes, and monocyte-derived macrophages. Via negative regulation of nuclear factor-kappa B and activator protein-1 signalling pathways, PPAR alpha inhibits expression of inflammatory genes, such as interleukin-6, cyclooxygenase-2, and endothelin-1. Furthermore, PPAR alpha inhibits expression of monocyte-recruiting proteins such as vascular cell adhesion molecule (VCAM)-1 and induces apoptosis in monocyte-derived macrophages. PPAR gamma activation in macrophages and foam cells inhibits the expression of activated genes such as inducible nitric oxide synthase, matrix metalloproteinase-9 and scavenger receptor A. PPAR gamma may also affect the recruitment of monocytes in atherosclerotic lesions as it is involved in the expression of VCAM-1 and intracellular adhesion molecule-1 in vascular endothelial cells. The involvement of PPAR in atherosclerosis, a disease with a chronic inflammatory character, suggests that they may play a role in other inflammatory-related diseases as well. PMID- 11007964 TI - Geranylgeranylated RhoB is sufficient to mediate tissue-specific suppression of Akt kinase activity by farnesyltransferase inhibitors. AB - Farnesyltransferase inhibitors (FTIs) induce apoptosis by elevating the levels of geranylgeranylated RhoB (RhoB-GG) in cells. However, the mechanism by which RhoB GG acts is unclear. Here we report that RhoB-GG is sufficient to mediate the suppressive effects of FTIs on the activity of the survival kinase Akt-1 in epithelial cells. This mechanism is tissue-specific insofar as it does not operate in fibroblasts. We discuss how the cell survival functions of RhoB and Akt may be linked biochemically in certain cell types. PMID- 11007965 TI - AMP-activated protein kinase counteracted the inhibitory effect of glucose on the phosphoenolpyruvate carboxykinase gene expression in rat hepatocytes. AB - The effect of AMP-activated protein kinase (AMPK) in the regulation of the phosphoenolpyruvate carboxykinase (PEPCK) gene expression was studied in isolated rat hepatocytes. Activation of AMPK by AICAR counteracted the inhibitory effect of glucose on the PEPCK gene expression, both at the mRNA and the transcriptional levels. It is proposed that a target for AMPK is involved in the inhibitory effect of glucose on PEPCK gene transcription. PMID- 11007966 TI - Desethylamiodarone interferes with the binding of co-activator GRIP-1 to the beta 1-thyroid hormone receptor. AB - Ligand binding to the thyroid hormone nuclear receptor beta1 (TRbeta(1)) is inhibited by desethylamiodarone (DEA), the major metabolite of the widely used anti-arrhythmic drug amiodarone. Gene expression of thyroid hormone (triiodothyronine, T(3))-regulated genes can therefore be affected by amiodarone due to less ligand binding to the receptor. Previous studies have indicated the possibility of still other explanations for the inhibitory effects of amiodarone on T(3)-dependent gene expression, probably via interference with receptor/co activator and co-repressor complex. The binding site of DEA is postulated to be on the outside surface of the receptor protein overlapping the regions where co activator and co-repressor bind. Here we show the effect of a drug metabolite on the interaction of TRbeta(1) with the co-activator GRIP-1 (glucocorticoid receptor interacting protein-1). The T(3)-dependent binding of GRIP-1 to the TRbeta(1) is disrupted by DEA. A DEA dose experiment showed that the drug metabolite acts like an antagonist under 'normal' conditions (at 10(-7) M T(3) and 5x10(-6)-->10(-3) M DEA), but as an agonist under extreme conditions (at 0 and 10(-9) M T(3) and >10(-4) M DEA). To our knowledge, these results show for the first time that a metabolite of a drug which was not devised for this purpose can interfere with nuclear receptor/co-activator interaction. PMID- 11007967 TI - A novel role for protein phosphatase 2A in the dopaminergic regulation of Na,K ATPase. AB - Stimulation of dopaminergic type 1 (D(1)) receptors increases lung edema clearance by regulating Na,K-ATPase function in the alveolar epithelium. We studied the role of serine/threonine protein phosphatases in the Na,K-ATPase regulation by D(1) agonists in A549 cells. We found that low doses of the type 1/2A protein phosphatase inhibitor okadaic acid as well as SV40 small t antigen transiently transfected into A549 cells prevented the D(1) agonist-induced increase in Na,K-ATPase activity and translocation from intracellular pools to the plasma membrane. This was associated with a rapid and transient increase in protein phosphatase 2A activity. We conclude that D(1) stimulation regulates Na,K ATPase activity by promoting recruitment of Na,K-ATPases from intracellular pools into the basolateral membranes of A549 cells via a type 2A protein phosphatase. PMID- 11007969 TI - Elevated expression of membrane type 1 metalloproteinase (MT1-MMP) in reactive astrocytes following neurodegeneration in mouse central nervous system. AB - Reactive astrocytes occurring in response to neurodegeneration are thought to play an important role in neuronal regeneration by upregulating the expression of extracellular matrix (ECM) components as well as the ECM degrading metalloproteinases (MMPs). We examined the mRNA levels and cellular distribution of membrane type matrix metalloproteinase 1 (MT1-MMP) and tissue inhibitors 1-4 of MMPs (TIMPs) in brain stem and spinal cord of wobbler (WR) mutant mice affected by progressive neurodegeneration and astrogliosis. MT1-MMP, TIMP-1 and TIMP-3 mRNA levels were elevated, whereas TIMP-2 and TIMP-4 expression was not affected. MT1-MMP was expressed in reactive astrocytes of WR. In primary astrocyte cultures, MT1-MMP mRNA was upregulated by exogeneous tumor necrosis factor alpha. Increased plasma membrane and secreted MMP activities were found in primary WR astrocytes. PMID- 11007968 TI - Functional involvement of a deoxy-D-xylulose 5-phosphate reductoisomerase gene harboring locus of Synechococcus leopoliensis in isoprenoid biosynthesis. AB - The present work aimed to proof the functionality of the non-mevalonate pathway in cyanobacteria. It was intended to isolate the 1-deoxy-D-xylulose 5-phosphate (DXP) reductoisomerase gene (dxr), as this gene encodes the enzyme which catalyzes a pathway-specific, indicative step of this pathway. For this purpose, a segment of dxr was amplified from Synechococcus leopoliensis SAUG 1402-1 DNA via PCR using oligonucleotides for conserved regions. Subsequent hybridization screening of a genomic cosmid library of S. leopoliensis with the PCR segment led to the identification of a 26. 5 kbp locus on which a dxr homologous gene and two adjacent open reading frames organized in one operon were localized by DNA sequencing. The functionality of the gene was demonstrated expressing the gene in Escherichia coli and using the purified gene product in a photometrical NADPH dependent test based on the substrate DXP generating system. While the content of one of the central intermediates of the isoprenoid biosynthesis (dimethylallyl diphosphate=DMADP) was significantly (P95% of the protein is stored within intracellular membranes. In response to insulin, Glut4 exhibits a large mobilisation to the plasma membrane. Studies of the amino acid motifs which regulate the unique trafficking of Glut4 have identified several key residues within the soluble cytoplasmic N- and C-terminal domains of Glut4. Of particular note is a Leu-498Leu-499 motif within the C-terminal domain that has been proposed to regulate both internalisation from the plasma membrane and sorting to an insulin-sensitive compartment. In this study, we have examined the role of the adjacent amino acids (Glu-491, Gln-492 and Glu-493) by their sequential replacement with Ala. Our results are consistent with the notion that Glu-491 and Glu-493 play an important role in the sub-endosomal trafficking of Glut4, as substitution of these residues with Ala results in increased levels of these proteins at the cell surface, reduced insulin-stimulated translocation and increased susceptibility to endosomal ablation. These residues, together with other identified sequences within the C-terminus of Glut4, are likely to be crucial targeting elements that regulate Glut4 subcellular distribution. PMID- 11007976 TI - Molecular identification of a glucose transporter from fish muscle. AB - In mammals and birds, several isoforms of facilitative glucose transporters have been identified (GLUT1-4), but no information is available regarding the molecules involved in glucose transport in other vertebrates. Here we report the cloning of a GLUT molecule from fish muscle with high sequence homology to GLUT4 and containing features characteristic of a functional GLUT. Fish GLUT is expressed predominantly in skeletal muscle, kidney and gill, which are tissues with known high glucose utilization. These results indicate that fish GLUT is structurally, and perhaps functionally, similar to the other known GLUTs expressed in muscle in mammalian and avian species. PMID- 11007977 TI - Selective stimulation of somatostatin receptor subtypes: differential effects on Ras/MAP kinase pathway and cell proliferation in human neuroblastoma cells. AB - In previous studies we have showed that somatostatin (SST) inhibits cell division, mitogen-activated protein (MAP) kinase and Ras activity in the human neuroblastoma cell line SY5Y. In the present study, we have assessed the role of a series of SST analogs, three of which were selective for SSTR1, SSTR2 or SSTR5, in these cellular events. All the analogs inhibited forskolin-induced cAMP accumulation. Selective stimulation of SSTR1 or SSTR2 but not of SSTR5 inhibited platelet-derived growth factor (PDGF)-induced [(3)H]thymidine incorporation. The three analogs inhibited PDGF-stimulated MAP kinase activity, at least at an early time. In contrast, none of the analogs used individually was able to inhibit PDGF stimulated Ras activity. A combined stimulation of SSTR2 and SSTR5 was necessary to obtain a significant inhibitory effect, suggesting the possibility of receptor heterodimerization. These results indicate that SST inhibition of Ras and MAP kinase activities takes place via different pathways and that SST inhibition of PDGF-induced cell proliferation occurs via a Ras-independent pathway. PMID- 11007978 TI - Cleavage of yeast tRNAPhe with complementary oligonucleotide conjugated to a small ribonuclease mimic. AB - An oligonucleotide conjugate bearing a chemical construct mimicking the catalytic center of ribonuclease A has been designed and studied. The conjugate efficiently cleaves yeast tRNAPhe at a single site adjacent to the target complementary sequence. PMID- 11007979 TI - Interaction of sigma 70 with Escherichia coli RNA polymerase core enzyme studied by surface plasmon resonance. AB - The interaction between the core form of bacterial RNA polymerases and sigma factors is essential for specific promoter recognition, and for coordinating the expression of different sets of genes in response to varying cellular needs. The interaction between Escherichia coli core RNA polymerase and sigma 70 has been investigated by surface plasmon resonance. The His-tagged form of sigma 70 factor was immobilised on a Ni2+-NTA chip for monitoring its interaction with core polymerase. The binding constant for the interaction was found to be 1.9x10(-7) M, and the dissociation rate constant for release of sigma from core, in the absence of DNA or transcription, was 4x10(-3) s(-1), corresponding to a half-life of about 200 s. PMID- 11007980 TI - MECP2 mutation in male patients with non-specific X-linked mental retardation. AB - In contrast to the preponderance of affected males in families with X-linked mental retardation, Rett syndrome (RTT) is a neurological disorder occurring almost exclusively in females. The near complete absence of affected males in RTT families has been explained by the lethal effect of an X-linked gene mutation in hemizygous affected males. We report here on a novel mutation (A140V) in the MECP2 gene detected in one female with mild mental retardation. In a family study, the A140V mutation was found to segregate in the affected daughter and in four adult sons with severe mental retardation. These results indicate that MECP2 mutations are not necessarily lethal in males and that they can be causative of non-specific X-linked mental retardation. PMID- 11007981 TI - Characterization of the active site thiol group of rhinovirus 2A proteinase. AB - Picornains 2A are cysteine proteases of picornaviruses, a virus family containing several human and animal pathogens. The pH dependencies of the alkylations of picornain 2A of rhinovirus type 2 with iodoacetamide and iodoacetate show two reactive thiol forms, namely the free thiolate ion at high pH and an imidazole assisted thiol group at low pH. Kinetic deuterium isotope effects do not support general base catalysis by the imidazole group, but rather the existence of a catalytically competent thiolate-imidazolium ion-pair. The nature of the ion-pair differs from that of papain, the paradigm of cysteine proteases. The ion-pair is confined to the same, unusually narrow pH range in which the enzyme exhibits catalytic activity. PMID- 11007983 TI - Nitric oxide synthases catalyze superoxide formation. PMID- 11007982 TI - Functional identification of the glycerol permease activity of Arabidopsis thaliana NLM1 and NLM2 proteins by heterologous expression in Saccharomyces cerevisiae. AB - NLM proteins (NOD26-like major intrinsic proteins) from plants contain amino acid sequence signatures which can be found in aquaporins including plant plasma membrane intrinsic proteins and tonoplast intrinsic proteins and glycerol permeases such as the Escherichia coli GlpF and the yeast FPS1 proteins. Heterologous expression of two members of the NLM subgroup from Arabidopsis thaliana (AtNLM1 and AtNLM2) in baker's yeast demonstrated the glycerol permease activity in addition to the previously described aquaporin activity of AtNLM1. The transport was non-saturable up to 100 mM extracellular glycerol concentration. Longer-chain sugar alcohols did not compete with the transport of radiolabelled glycerol and hexoses were also not transported through the pore. PMID- 11007984 TI - Clonidine induces upper airway closure in awake goats. AB - We examined the effects of the alpha(2)-adrenoceptor (alpha(2)-AR) agonist clonidine on pressure-flow relationships in the upper airway. Inspired and expired airflows, subglottic tracheal pressure (PTR), mask pressure and middle pharyngeal constrictor (MPC) and diaphragm electromyogram (EMG) activities were recorded in awake standing goats. Clonidine-induced central apneas were always associated with continuous tonic activation of the MPC. Subglottic PTR during expiration increased progressively in a dose-dependent manner after clonidine administration. In all cases, positive subglottic PTR was maintained throughout the duration of clonidine-induced apneas and was sufficient to retard or prevent expiratory flow during early and mid-expiration indicating complete airway closure. The effects of clonidine were reversed by selective alpha(2)-AR blockade with SKF-86466. Central apneas after spontaneous augmented breaths (sighs) were associated with continuous tonic activation of the MPC throughout the duration of the prolonged TE intervals. However, subglottic PTR during expiration was not significantly different from control breaths and there was no evidence of increased expiratory airway resistance or delayed expiratory flow. We conclude that continuous tonic activation of pharyngeal adductor muscles appears to be a constant feature of central apnea in the awake goat independent of the initiating cause of the apnea. However, our data suggest that MPC activation alone may not be sufficient to cause complete closure of the upper airway during central apnea. PMID- 11007986 TI - Developmental changes in cardio-respiratory responses to hypoxia and hypercapnia in anesthetized low-birth-weight rats. AB - The present study compared the developmental changes in the cardio-respiratory responses to hypoxia and hypercapnia between full-term low-birth-weight (LBW) and control rats during the postnatal period. The heart rate (HR), respiratory frequency (fR) and amplitude (aR) were measured during hypoxia (10% O(2) for 10 min) and hypercapnia (5% CO(2) for 10 min) in rats aged 7, 14 and 21 days, anesthetized with urethane. During hypoxia, HR was not significantly modified in the younger rats of both groups. In the older rats, aged 14 and 21 days, HR was markedly diminished, with a more pronounced decrease in LBW rats. The HR recovery was never observed in the older LBW rats. The fR and aR showed an age-related increase in both groups: a biphasic fR pattern observed on day 7 was replaced by a sustained increase on days 14 and 21. In contrast to controls, LBW rats never displayed a fR recovery during reoxygenation. In controls, aR shifted from a biphasic pattern in the younger rats to a sustained increase in the older ones. The LBW rats only displayed a decrease of aR in the younger, while in the older ones, a transient and slight increase preceded this decrease. During hypercapnia, the only significant difference detected between these two groups was that aR increased in LBW rats to a greater extent than in controls on days 14 and 21. Altogether, our results revealed a markedly attenuated cardio-respiratory response to hypoxia in LBW rats, but no such effect in response to hypercapnia. PMID- 11007985 TI - Changes in dopamine D(2)-receptor modulation of the hypoxic ventilatory response with chronic hypoxia. AB - Modulation of the hypoxic ventilatory response (HVR) by dopamine D(2)-receptors (D(2)-R) in the carotid body (CB) and central nervous system (CNS) are hypothesized to contribute to ventilatory acclimatization to hypoxia. We tested this with blockade of D(2)-R in the CB or CNS in conscious rats after 0, 2 and 8 days of hypoxia. On day 0, CB D(2)-R blockade significantly increased VI and frequency (fR) in hyperoxia (FI(O(2))=0.30), but not hypoxia (FI(O(2))=0.10). CNS D(2)-R blockade significantly decreased fR in hypoxia only. On day 2, neither CB nor CNS D(2)-R blockade affected VI or fR. On day 8, CB D(2)-R blockade significantly increased hypoxic VI and fR. CNS D(2)-R blockade significantly decreased hypoxic VI and fR. CB and CNS D(2)-R modulation of the HVR decreased after 2 days of hypoxia, but reappeared after 8 days. Changes in the opposing effects of CB and CNS D(2)-R on the HVR during chronic hypoxia cannot completely explain ventilatory acclimatization in rats. PMID- 11007987 TI - Characterizations of eupnea, apneusis and gasping in a perfused rat preparation. AB - In vivo mammalian preparations can exhibit eupnea, apneusis and gasping. In vitro mammalian preparations exhibit only a single invariant pattern, which appears identical to gasping. We characterized the patterns of ventilatory activity of a perfused heart-brainstem preparation of the juvenile rat. In this preparation, phrenic activity has a 'ramp-like' rise similar to eupnea in vivo. Peak phrenic activity declines and ultimately disappears in hypocapnia. In hypercapnia, both frequency and peak of phrenic bursts increase. In hypoxia, such increases are transient. The phrenic burst is terminated by electrical stimulation of the pontile 'pneumotaxic center' and, as in apneusis, is prolonged by lesions in this region. With severe hypoxia or ischemia, the 'ramp-like' phrenic activity is replaced by the 'decrementing' pattern of gasping. Variables of phrenic activity in gasping produced in hypoxia and ischemia are identical. We conclude that the perfused juvenile rat preparation exhibits patterns of eupnea, apneusis and gasping which are similar to in vivo mammalian preparations. PMID- 11007988 TI - Sensitivities of eupnea and gasping to alterations in temperature of in vivo and perfused rat preparations. AB - Severe hypoxia or ischemia causes an alteration from eupnea to gasping. At body temperatures approximating 37 degrees C in vivo, the frequency of gasping is much less than that of eupnea. However in a perfused juvenile rat preparation, which is maintained at 30-31 degrees C, the frequency of eupnea and gasping is the same. We hypothesized that brainstem mechanisms responsible for the neurogenesis of eupnea and gasping might have different sensitivities to alterations in temperature. In both decerebrate adult rats in vivo and in a perfused juvenile rat preparation, eupnea and gasping had different frequencies at rectal or perfusate temperatures in excess of 34 degrees C, whereas, at lower temperatures, eupnea and gasping had identical frequencies in both preparations. These findings support the conclusion that different brainstem mechanisms underlie the neurogenesis of eupnea and gasping. In addition, these findings have implications for interpretation of results from in vitro mammalian preparations, which are examined at temperatures at which the frequency of eupnea and gasping are indistinguishable. PMID- 11007989 TI - Estimation of diaphragm length in patients with severe chronic obstructive pulmonary disease. AB - In patients with advanced chronic obstructive pulmonary disease (COPD) diaphragm function may be compromised because of reduced muscle fibre length. Diaphragm length (L(Di)) can be estimated from measurements of transverse diameter of the rib cage (D(Rc)) and the length of the zone of apposition (L(Zapp)) in healthy subjects, but this method has not been validated in patients with COPD. Postero anterior chest radiographs were obtained at total lung capacity (TLC), functional residual capacity (FRC) and residual volume (RV) in nine male patients with severe COPD (mean [S.D.]; FEV(1), 23 [6] %pred.; FRC, 199 [15] %pred.). Radiographs taken at TLC were used to identify the lateral costal insertions of the diaphragm (L(Zapp) assumed to approach zero at TLC). L(Di) was measured directly and also estimated from measurements of L(Zapp) and D(Rc) using a prediction equation derived from healthy subjects. The estimation of L(Di) was highly accurate with an intraclass correlation coefficient of 0.93 and 95% CI of approximately +/-8% of the true value. L(Di) decreased from 426 (64) mm at RV to 305 (31) mm at TLC. As there were only small and variable changes in D(Rc) across the lung volume range, most of the L(Di) changes occurred in the zone of apposition. Additional studies showed that measurements of L(Di) from PA and lateral radiographs performed at different lung volumes were tightly correlated. These results suggest that non-invasive measurements of L(Zapp) in the coronal plane (e.g. using ultrasonography) and D(Rc) (e.g. using magnetometers) can be used to provide an accurate estimate of L(Di) in COPD patients. PMID- 11007990 TI - Arterial hypoxaemia in endurance athletes is greater during running than cycling. AB - The effect of both training discipline and exercise modality on exercise-induced hypoxaemia (EIH) was examined in seven runners and six cyclists during 5 min high intensity treadmill and cycle exercise. There were no significant interactions between training discipline, exercise modality and arterial P(O(2)) (Pa(O(2))) when subject groups were considered separately but when pooled there were significant differences between exercise modalities. After min 2 of exercise arterial hydrogen ion concentration, minute ventilation, alveolar P(O(2)) (PA(O(2))) and Pa(O(2)) were all lower with treadmill running with the largest differential for the latter occurring at min 5 (treadmill, 80.8+/-1.8; cycle, 90.2+/-2.5, mmHg, N=13, P< or = 0.05). At every min of exercise, the differences in Pa(O(2)) between the ergometers were strongly associated with similar differences in PA(O(2)) and alveolar to arterial P(O(2)) (PA(O(2))-Pa(O(2))). It is concluded that the greater EIH with treadmill running is a consequence of the combined effect of a reduced lactic acidosis-induced hyperventilation and greater ventilation-perfusion inequality with this exercise mode. PMID- 11007991 TI - Preface PMID- 11007992 TI - Dinucleoside polyphosphates-friend or foe? AB - Despite being known for over 30 years, the functions of the dinucleoside polyphosphates, such as diadenosine 5',5"'-P(1), P(4)-tetraphosphate (Ap(4)A) and diadenosine 5',5"'-P(1), P(3)-triphosphate (Ap(3)A), are still unclear. On the one hand, they may have important signalling functions, both inside and outside the cell (friend), while on the other hand, they may simply be the unavoidable by products of certain biochemical reactions, which, if allowed to accumulate, would be potentially toxic through their structural similarity to ATP and other essential mononucleotides (foe). Here, the occurrence, synthesis, degradation, and proposed functions of these compounds are briefly reviewed, along with some new data and recent evidence supporting roles for Ap(3)A and Ap(4)A in the cellular decision making processes leading to proliferation, quiescence, differentiation, and apoptosis. Hypotheses are forwarded for the involvement of Ap(4)A in the intra-S phase DNA damage checkpoint and for Ap(3)A and the pFhit (fragile histidine triad gene product) protein in tumour suppression. It is concluded that the roles of friend and foe are not incompatible, but are distinguished by the concentration range of nucleotide achieved under different circumstances. PMID- 11007993 TI - Synthesis of dinucleoside polyphosphates catalyzed by firefly luciferase and several ligases. AB - The findings presented here originally arose from the suggestion that the synthesis of dinucleoside polyphosphates (Np(n)N) may be a general process involving enzyme ligases catalyzing the transfer of a nucleotidyl moiety via nucleotidyl-containing intermediates, with release of pyrophosphate. Within this context, the characteristics of the following enzymes are presented. Firefly luciferase (EC 1.12. 13.7), an oxidoreductase with characteristics of a ligase, synthesizes a variety of (di)nucleoside polyphosphates with four or more inner phosphates. The discrepancy between the kinetics of light production and that of Np(n)N synthesis led to the finding that E*L-AMP (L = dehydroluciferin), formed from the E*LH(2)-AMP complex (LH(2) = luciferin) shortly after the onset of the reaction, was the main intermediate in the synthesis of (di)nucleoside polyphosphates. Acetyl-CoA synthetase (EC 6.2.1.1) and acyl-CoA synthetase (EC 6.2.1. 8) are ligases that synthesize p(4)A from ATP and P(3) and, to a lesser extent, Np(n)N. T4 DNA ligase (EC 6.5.1.1) and T4 RNA ligase (EC 6.5.1.3) catalyze the synthesis of Np(n)N through the formation of an E-AMP complex with liberation of pyrophosphate. DNA is an inhibitor of the synthesis of Np(n)N and conversely, P(3) or nucleoside triphosphates inhibit the ligation of a single strand break in duplex DNA catalyzed by T4 DNA ligase, which could have therapeutic implications. The synthesis of Np(n)N catalyzed by T4 RNA ligase is inhibited by nucleoside 3'(2'),5'-bisphosphates. Reverse transcriptase (EC 2.7.7.49), although not a ligase, catalyzes, as reported by others, the synthesis of Np(n)ddN in the process of removing a chain termination residue at the 3'-OH end of a growing DNA chain. PMID- 11007995 TI - Specific and nonspecific enzymes involved in the catabolism of mononucleoside and dinucleoside polyphosphates. AB - This review concerns enzymes that can degrade nucleoside 5'-tetra- and pentaphosphates (p(4)N and p(5)N) and those that can degrade various dinucleoside polyphosphates (Np(3-6)N'). Most of these enzymes are hydrolases, and they occur in all types of organisms. Certain fungi and protozoa also possess specific Np(n)N' phosphorylases. Specific p(4)N hydrolases have been demonstrated in mammals and in plants. In yeast, p(4)N and p(5)N are hydrolyzed by exopolyphosphatases. Among other hydrolases that can degrade these minor mononucleotides are phosphatases, apyrase, and (asymmetrical) Np(4)N' hydrolase, as well as the nonspecific adenylate deaminase. Np(n)N's are good substrates for Type I phosphodiesterases and nucleotide pyrophosphatases, and diadenosine polyphosphates are easily deaminated to diinosine polyphosphates by nonspecific adenylate deaminases. Specific Np(3)N' hydrolases occur in both prokaryotes and eukaryotes. Interestingly, the human fragile histidine triad (Fhit) tumor suppressor protein appears to be a typical Np(3)N' hydrolase. Among the specific Np(4)N' hydrolases are asymmetrically cleaving ones, which are typical of higher eukaryotes, and symmetrically cleaving enzymes found in Physarum polycephalum and in many bacteria. An enzyme that hydrolyzes both diadenosine tetraphosphate and diadenosine triphosphate has been found in the fission yeast Schizosaccharomyces pombe. Its amino acid sequence is similar to that of the human Fhit/Np(3)N' hydrolase. Very recently, a typical (asymmetrical) Np(4)N' hydrolase has been demonstrated for the first time in a bacterium-the pathogenic Bartonella bacilliformis. Another novelty is the discovery of diadenosine 5', 5"'-P(1),P 6 hexaphosphate hydrolases in budding and fission yeasts and in mammalian cells. These enzymes and the (asymmetrical) Np(4)N' hydrolases have the amino acid motif typical of the MutT (or Nudix hydrolase) family. In contrast, the Schizosaccharomyces pombe Ap(4)A/Ap(3)A hydrolase, the human Fhit protein, and the yeast Np(n)N' phosphorylases belong to a superfamily GAFH, which includes the histidine triad proteins. PMID- 11007994 TI - Diadenosine polyphosphate receptors. from rat and guinea-pig brain to human nervous system. AB - Diadenosine polyphosphates are a family of naturally occurring nucleotidic compounds present in secretory vesicles together with other chemical messengers. The exocytotic release of these compounds permits them to stimulate receptors termed "purinoceptors" or "ATP receptors." Purinoceptors for nucleotides are named P2 in contrast with those sensitive to nucleosides (P1). P2 receptors are further subdivided into metabotropic P2Y receptors, further divided into 5 subtypes, and ionotropic P2X receptors, with 7 different subtypes. Diadenosine polyphosphates can activate recombinant P2Y(1), P2Y(2), and P2Y(4) and recombinant homomeric P2X(1), P2X(2), P2X(3), P2X(4), and P2X(6). Heteromeric P2X receptors change their sensitivity to diadenosine polyphosphates when co-assembly between different subunits occurs. Diadenosine polyphosphates can activate specific receptors termed dinucleotide receptors or P4 receptors, which are insensitive to other nucleosides or nucleotides. The P4 receptor is a receptor operated Ca(2)+ channel present in rat brain synaptic terminals, stimulated by diadenosine pentaphosphate and diadenosine tetraphosphate. This receptor is strongly modulated by protein kinases A and C and protein phosphatases. The dinucleotide receptor is present in different brain areas, such as midbrain (in rat and guinea-pig), cerebellum (in guinea-pig), and cortex (in human). PMID- 11007996 TI - The role of adenosine in preconditioning. AB - Preconditioning is a powerful form of (myocardial) protection that follows brief sublethal ischemia. G-protein-coupled receptors constitute the trigger for entrance to the preconditioned state. In conjunction with other receptors, various membrane adenosine receptors play an important role in the transduction of extracellular signals, leading to protection by preconditioning, lasting 1-3 hr. Adenosine A(1)- and A(3)-receptors mediate inhibition of adenylate cyclase via a guanine nucleotide binding inhibitory protein (G(i/o)). A(2)-receptors couple to a comparable stimulatory protein (G(s)). Adenosine receptors are especially abundant in the central nervous system; in lesser numbers, they are found in many tissues, including the heart. A(1)-receptors are located on cardiomyocytes and vascular smooth muscle cells, A(2)-receptors on endothelial and vascular smooth muscle cells, and A(3)-receptors on ventricular myocytes. Ischemic preconditioning by endogenous adenosine takes place through A(1)- and A(3)-receptors. A(2A/B)-receptor activation results in vasodilation. The relevance of cellular mediators, such as 5'-nucleotidase, to generate adenosine for preconditioning is controversial. In contrast, the role of protein kinase C (PKC) is clearly established. Signals from different receptors converge at PKC, reaching a threshold activation of the kinase necessary to induce protection. Tyrosine and mitogen-activated protein kinases may play a role in addition to PKC. The exact products downstream responsible for the memory of preconditioning are elusive. A prime candidate for the end-effector of preconditioning is the K(ATP) channel. Preconditioning with adenosine-receptor agonists offers the possibility for treatment of coronary artery disease, but research in this field is still in its infancy. PMID- 11007997 TI - The adenosine-induced mechanism for the acquisition of ischemic tolerance in primary rat neuronal cultures. AB - Neurons can be preconditioned by various procedures to resist ischemic insult. The preconditioning mechanism induced by adenosine ("the adenosine mechanism") was characterized in primary rat neuronal cultures, employing a model of chemical ischemia. The protective mechanism, initiated by activation of adenosine receptors, consists of a signal transduction pathway, involving activation of protein kinase C (PKC) and opening of ATP-sensitive potassium (K(ATP)) channels. Direct activation (and inhibition) of PKC, as well as opening of K(ATP) channels, also confers protection. The opening of the K(ATP) channels mediates the signal activated by the adenosine receptors, and probably also that activated by PKC. The acquired ischemic resistance lasts up to 5 days, depending on the activating substance. The adenosine-activated cascade of events leading to ischemic tolerance in neurons is similar to that operating in cardiomyocytes. PMID- 11007998 TI - Tumor-promoting functions of adenosine. AB - Tumor growth is a multifactorial process that, in addition to mutations leading to dysregulated expression of oncogenes and tumor suppressive genes, requires specific conditions that provide a supportive physiological environment at the primary and metastatic sites of the disease. Adenosine is one of the factors potentially contributing to tumor growth that thus far has not received adequate attention, despite evidence for a broad range of cytoprotective, growth promoting, and immunosuppressive activities. Adenosine accumulates in solid tumors at high concentrations, and has been shown to stimulate tumor growth and angiogenesis and to inhibit cytokine synthesis, adhesion of immune cells to the endothelial wall, and the function of T-cells, macrophages, and natural killer cells. However, the mechanisms whereby adenosine accumulates in cancer and the specific effects that result from this accumulation are not well understood. This article surveys the available evidence that supports an important role of adenosine in cancer. PMID- 11008000 TI - Phosphorylation of nucleosides and nucleoside analogs by mammalian nucleoside monophosphate kinases. AB - Nucleoside monophosphate kinases catalyze the reversible phosphotransferase reaction between nucleoside triphosphates and monophosphates, i.e., monophosphates are converted to their corresponding diphosphate form. These enzymes play an important role in the synthesis of nucleotides that are required for a variety of cellular metabolic processes, as well as for RNA and DNA synthesis. Human tissues contain a thymidylate kinase, a uridylate-cytidylate kinase, five isozymes of adenylate kinase, and several guanylate kinases. Nucleoside monophosphate kinases are also required for the pharmacological activation of therapeutic nucleoside and nucleotide analogs. This overview is focused on the substrate specificity, tissue distribution, and subcellular location of the mammalian monophosphate kinases and their role in the activation of nucleoside and nucleotide analogs. PMID- 11007999 TI - Effect of antimetabolite drugs of nucleotide metabolism on the anti-human immunodeficiency virus activity of nucleoside reverse transcriptase inhibitors. AB - A number of attempts are currently underway to combine antimetabolite drugs of nucleotide metabolism with a nucleoside reverse transcriptase inhibitor (NRTI) targeting human immunodeficiency virus (HIV) to improve the antiviral efficacy of the NRTIs and to better control HIV drug resistance. Hydroxyurea, a ribonucleotide reductase inhibitor, is currently combined with the NRTI didanosine (2',3'-dideoxyinosine) in clinical trials. However, other cellular target enzymes, including thymidylate synthase, inosinate dehydrogenase, cytidine 5'-triphosphate synthetase, and other enzymes from the de novo nucleotide biosynthesis pathway, can also be considered to potentiate the antiviral action of NRTIs. The underlying reasons for the potentiation of the antiviral activity of the NRTIs by antimetabolite drugs of nucleotide metabolism can be multiple. Decreased endogenous 2'-deoxynucleoside-5'-triphosphate (dNTP) pools result in a better competition of the NRTI (as its triphosphate derivative), with the dNTPs for the virus-encoded reverse transcriptase to be recognized as a substrate for the DNA polymerization reaction and subsequently to be incorporated into the growing viral DNA chain. Also, an increased metabolism (phosphorylation) of the NRTI by stimulatory enzyme feedback mechanisms may result in the production of higher levels of NRTI triphosphate. Thus, higher intracellular ratios of NRTI triphosphate/dNTP created by well-defined combinations of NRTIs and antimetabolite drugs enable a more profound inhibitory effect of the NRTI against the reverse transcriptase (and thus, against the virus) and a better suppression of resistant (mutant) virus strains. A profound evaluation of this relatively new concept in the clinical setting will reveal whether this approach will establish a place in future treatment modalities of HIV infections. PMID- 11008001 TI - Cyclic nucleotide analogs as biochemical tools and prospective drugs. AB - Cyclic AMP (cAMP) and cyclic GMP (cGMP) are key second messengers involved in a multitude of cellular events. From the wealth of synthetic analogs of cAMP and cGMP, only a few have been explored with regard to their therapeutic potential. Some of the first-generation cyclic nucleotide analogs were promising enough to be tested as drugs, for instance N(6),O(2)'-dibutyryl-cAMP and 8-chloro-cAMP (currently in clinical Phase II trials as an anticancer agent). Moreover, 8-bromo and dibutyryl analogs of cAMP and cGMP have become standard tools for investigations of biochemical and physiological signal transduction pathways. The discovery of the Rp-diastereomers of adenosine 3',5'-cyclic monophosphorothioate and guanosine 3',5'-cyclic monophosphorothioate as competitive inhibitors of cAMP and cGMP-dependent protein kinases, as well as subsequent development of related analogs, has proven very useful for studying the molecular basis of signal transduction. These analogs exhibit a higher membrane permeability, increased resistance against degradation, and improved target specificity. Furthermore, better understanding of signaling pathways and ligand/protein interactions has led to new therapeutic strategies. For instance, Rp-8-bromo-adenosine 3',5' cyclic monophosphorothioate is employed against diseases of the immune system. This review will focus mainly on recent developments in cyclic nucleotide-related biochemical and pharmacological research, but also highlights some historical findings in the field. PMID- 11008002 TI - Basis for effective combination cancer chemotherapy with antimetabolites. AB - Most current chemotherapy regimens for cancer consist of empirically designed combinations, based on efficacy and lack of overlapping toxicity. In the development of combinations, several aspects are often overlooked: (1) possible metabolic and biological interactions between drugs, (2) scheduling, and (3) different pharmacokinetic profiles. Antimetabolites are used widely in chemotherapy combinations for treatment of various leukemias and solid tumors. Ideally, the combination of two or more agents should be more effective than each agent separately (synergism), although additive and even antagonistic combinations may result in a higher therapeutic efficacy in the clinic. The median-drug effect analysis method is one of the most widely used methods for in vitro evaluation of combinations. Several examples of classical effective antimetabolite-(anti)metabolite combinations are discussed, such as that of methotrexate with 6-mercaptopurine or leucovorin in (childhood) leukemia and 5 fluorouracil (5FU) with leucovorin in colon cancer. More recent combinations include treatment of acute-myeloid leukemia with fludarabine and arabinosylcytosine. Other combinations, currently frequently used in the treatment of solid malignancies, include an antimetabolite with a DNA-damaging agent, such as gemcitabine with cisplatin and 5FU with the cisplatin analog oxaliplatin. The combination of 5FU and the topoisomerase inhibitor irinotecan is based on decreased repair of irinotecan-induced DNA damage. These combinations may increase induction of apoptosis. The latter combinations have dramatically changed the treatment of incurable cancers, such as lung and colon cancer, and have demonstrated that rationally designed drug combinations offer new possibilities to treat solid malignancies. PMID- 11008003 TI - Delivery systems for antisense oligonucleotides. AB - In vitro, the efficacy of the antisense approach is strongly increased by systems delivering oligodeoxyribonucleotides (ODNs) to cells. Up to now, most of the developed vectors favor ODN entrance by a mechanism based on endocytosis. Such is the case for particulate systems, including liposomes (cationic or non-cationic), cationic polyelectrolytes, and delivery systems targeted to specific receptors. Under these conditions, endosomal compartments may represent a dead end for ODNs. Current research attempts to develop conditions for escaping from these compartments. A new class of vectors acts by passive permeabilization of the plasma membrane. It includes peptides, streptolysin O, and cationic derivatives of polyene antibiotics. In vivo, the interest of a delivery system, up to now, has appeared limited. Development of vectors insensitive to the presence of serum seems to be a prerequisite for future improvements. PMID- 11008004 TI - Towards an understanding of the functional significance of N-terminal domain divergence in human AMP deaminase isoforms. AB - Human AMP deaminase (AMPD; EC 3.5.4.6) isoforms are encoded by a multigene family and have conserved C-terminal domains that contain catalytic residues and an ATP binding site. N-terminal domains diverge dramatically, yet are conserved when compared across mammalian species. Cross-species conservation of entire gene specific polypeptides (e.g., rat versus human AMPD1) suggests that divergent N terminal domains may play a role in isoform-specific properties of the enzyme. It now has become evident that the majority of published data used to characterize purified AMPD isoforms were likely derived from preparations lacking significant portions of their N-terminal domains (up to nearly 100 residues). Accumulating evidence indicates that divergent N-terminal sequences do influence catalytic behavior, protein-protein interactions, and intracellular distributions of this enzyme. PMID- 11008005 TI - Five year survival and later outcome of patients with X-ray occult lung cancer detected by sputum cytology. AB - BACKGROUND: A cohort of 51 consecutive patients with roentgenographically occult lung cancer, identified by sputum cytology and confirmed by bronchoscopy was reported previously. METHODS: All patients have now been followed beyond 5 years and the causes of death ascertained. RESULTS: The actual 5-year survival of 27 patients who were resected for cure was 74% including death for all causes. The 5 year survival of all patients who received either surgery or radiation in an attempt to cure was 54.3%. Twelve secondary cancers were found by sputum cytology; eight of these patients have died. CONCLUSIONS: Sputum cytology can be useful in the identification of early stage lung cancer in patients at high-risk where the chances of cure are favorable. PMID- 11008006 TI - Characteristics of lung cancer in Korea, 1997. AB - The high proportion of smokers and the incidence of advanced, unresectable lung cancer in Korea were examined to aid the development of a national anti-smoking program and the early detection of lung cancer. Koreans are a single racial group with a high smoking rate among men and a contrastingly low smoking rate among women. This report documents a retrospective investigation conducted by The Korean Academy of Tuberculosis and Respiratory Disease into the characteristics of all lung cancers diagnosed between 1 January 1997 and 31 December 1997 in Korea. Among the 3794 patients included in this study, 76.8% were smokers and, in particular, 89.8% of the males were smokers. Squamous cell carcinoma was the most frequent type of lung cancer encountered (44.7%), followed by adenocarcinoma (27.9%). The smoking rate in the case of adenocarcinoma was significantly lower than that found in both squamous cell carcinoma and small cell cancer. The most common symptom was a cough. Only 7.2% of patients were asymptomatic. Bronchoscopic biopsy has a primary role in the diagnosis of squamous cell carcinoma and small cell cancer, but percutaneous needle biopsy has a more important role in the case of adenocarcinoma. Two-thirds of the nonsmall cell lung cancer patients were detected in the unresectable advanced stages (IIIB and IV). In contrast to other countries, squamous cell carcinoma is still the most frequent type of lung cancer in Korea. The high proportion of smokers and the incidence of advanced, unresectable lung cancer at diagnosis have urged development of a national anti-smoking program to promote the cessation of smoking and the early detection of lung cancer. PMID- 11008007 TI - A systematic review of the role of etoposide and cisplatin in the chemotherapy of small cell lung cancer with methodology assessment and meta-analysis. AB - PURPOSE: Cisplatin (CDDP) and etoposide (VP16) are considered major standard cytotoxic drugs for small cell lung cancer (SCLC). The present systematic review had as its objective the evaluation of their role, as components of chemotherapy regimens, on survival. METHODS: Published randomised clinical trials (from 1980 to 1998) were selected comparing, in SCLC patients, chemotherapy regimens, given as first-line therapy. One arm (the experimental arm) had to include CDDP and/or VP16, while another had to omit the same drug(s). Trials quality was assessed by two published scores (Chalmers and European Lung Cancer Working Party (ELCWP)). For each individual trial, the hazard ratio (HR) of the survival distributions was estimated on the basis of reported statistics or, if not available, by extracting, from the survival graphical representations, the data required to construct the difference between expected and observed numbers of events as calculated in the log-rank statistic. A combined hazard ratio was obtained by the Peto method (a value < 1 meaning a benefit for CDDP and/or VP16). RESULTS: Thirty six trials eligible for our systematic review were identified, classified into four groups (I-IV): group I, 1 trial testing a CDDP-based regimen (without VP16) against another arm not including either CDDP or VP16; group II, 17 trials testing a VP16-based regimen (without CDDP) against a regimen without VP16 and CDDP; group III, nine trials comparing a regimen including CDDP and VP16 with a regimen using neither drug; and, finally, group IV, nine trials comparing a regimen based on both drugs with a regimen based on VP16 only. Overall, Chalmers and ELCWP scores correlated well (r(S) = 0.76, P < 0. 001) and had respective median scores of 50.3 and 63.7%. The number of eligible patients did not have a significant impact on the scores as well as the trials group, the trial positivity (a positive trial defined as showing itself a statistically significant survival benefit for the experimental regimen), overall or in categories, and the year of publication. Combined hazard ratios with 95% confidence intervals were: 0.70 (0.41-1.21) for group I, 0.72 (0.67-0.78) for II, 0.57 (0.51-0.64) for III, and 0.74 (0.66-0.83) for IV, showing a survival benefit in favour of regimens including etoposide alone or in combination with cisplatin, justifying with high significance levels the use of each of these drugs. Overall survival benefits could also be shown for regimens including CDDP (HR = 0.61; confidence interval (CI), 0.57-0.66), as well as for those including VP16 (HR = 0. 65; CI, 0.61-0.69). Robustness of these results has to be confirmed with appropriate randomised trials. PMID- 11008008 TI - Serum biomarkers of non-neuron-endocrine origin in small-cell lung cancer: a 16 year study on carcinoembryonic antigen, tissue polypeptide antigen and lactate dehydrogenase. AB - BACKGROUND: Biomarkers of non-neuron-endocrine origin are measured only occasionally in the sera of patients with small-cell lung cancer (SCLC). An exception to this rule is carcinoembryonic antigen (CEA), for which, however, there is no consistent evidence. Based on such a premise, we decided to review the Cuneo Lung Cancer Study Group 16-year-experience with non-neuron-endocrine markers in SCLC. METHODS: a total of 619 CEA, 621 tissue polypeptide antigen (TPA), and 616 lactate dehydrogenase (LDH) serum assays were obtained from 160 consecutive SCLC at diagnosis, during, and after treatment. Demographic, clinical, laboratory, and tumoral correlates were also available for another 25 pretreatment and 14 posttreatment variables. RESULTS: bivariate correlation analyses showed that LDH and TPA were significantly related to each other, and both of them were also correlated with disease extent, and treatment response. LDH correlation indexes were higher than that of TPA, especially those regarding the parameters of disease extent. CEA was correlated only with the category of treatment response. Receiver-operating characteristic (ROC) analysis confirmed the correlation between stage disease at diagnosis and both LDH (P = 0.000) and TPA (P = 0.002), while the treatment failure was better recognized by TPA (P = 0.000). In univariate analysis, both LDH and TPA were correlated with survival (P = 0.000 and 0.092, respectively); however, only LDH remained significant in multivariate analysis (P = 0.012). CONCLUSIONS: the evidence from this study does not suggest a routine CEA test in SCLC. LDH remains particularly useful and it should be kept in use. Finally, data on TPA is insufficient to advocate its systematic use in this type of malignancy. PMID- 11008009 TI - LDH or NSE or LDH and NSE as pretreatment prognostic factors in small cell lung cancer? A commentary. PMID- 11008010 TI - Mechanisms of the release of CYFRA21-1 in human lung cancer cell lines. AB - The CYFRA 21-1 assay which detects cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. However, the reason why some lung cancer cell lines release CK19 fragment in culture supernatants and others do not, remains unclear. It was hypothesized that the release of CK19 fragment may be elucidated by the expression of mRNA for CK19. In order to prove this, the mRNA for CK19 was quantitatively evaluated by the competitive reverse transcriptase-polymerase chain reaction (competitive RT-PCR). The level of CYFRA 21-1 in the culture supernatant was measured by an immunoradiometric assay. CK19 protein synthesis was evaluated by a Western blotting and immunohistochemistry. Fourteen lung cancer cell lines were evaluated, and the amount of mRNA correlated well with the level of CYFRA 21-1 in culture supernatants. Analysis of genomic DNA for CK19 demonstrated that three cell lines which could not produce CYFRA 21-1, conjectured that some abnormalities in exon 1 or the 5'-region upstream from exon 1. In conclusion, it was demonstrated that the release of CK19 fragment was closely related to the expression of mRNA for CK19, and the possibility that genomic change of CK19 DNA down-regulated the expression of mRNA for CK19 was suggested. PMID- 11008011 TI - The thin red line: angiogenesis in normal and malignant hematopoiesis. AB - This review describes the current knowledge about cell subsets involved in vasculogenesis (i.e., differentiation of endothelial cells from mesodermal precursors) and angiogenesis (i.e., blood vessel generation from pre-existing vessels), together with recent findings about angiogenesis and antiangiogenic therapies in hematopoietic malignancies such as leukemia, lymphoma, myeloma, and myelodysplastic syndromes. PMID- 11008012 TI - Granulocyte-macrophage colony-stimulating factor is not responsible for residual thrombopoiesis in mpl null mice. AB - OBJECTIVE: To examine the role of granulocyte-macrophage colony-stimulating factor (GM-CSF) in thrombopoiesis. MATERIALS AND METHODS: Thrombopoietin unresponsi ve mice (mpl null mice), which have a profound reduction in platelets and mature megakaryocytes, were interbred with mice that do not respond to GM-CSF or interleukin 5 (betac null mice), and hematopoiesis was examined. In initial experiments on a mixed genetic background, double mutant mice (betac/mpl null mice) showed an unexpected amelioration of the thrombocytopenia seen in mpl null mice. Platelet counts were elevated approximately twofold in betac/mpl null mice compared with mpl null mice (mpl null 73+/-31; betac/mpl null 164+/-70; n = 10 to 29 mice per genotype, p<0.00001). This was associated with lessening of the deficit of megakaryocytes, progenitor cells, and colony-forming units spleen seen in mpl null mice. This amelioration of the mpl null phenotype in betac/mpl null mice on a mixed genetic background was highly statistically significant. To determine whether this amelioration of phenotype was solely the consequence of loss of betac signaling, progeny of a second intercross on a C57BL/6 background (B6betac/mpl null mice) were examined. When the resulting B6betac/mpl null mice were analyzed and compared with B6mpl null littermates, the increase in platelet count, hematopoietic progenitor cell number, and colony-forming units spleen number was no longer observed. CONCLUSION: There was no additional effect seen as a result of loss of betac signaling. GM-CSF did not play a significant role in thrombopoiesis, even in combination with the absence of thrombopoietin signaling. These results highlight problems that can be encountered when studying introduced mutations in mice. They exemplify the importance of eliminating the influence of modifying genes when attributing biologic differences to specific introduced genetic alterations. PMID- 11008013 TI - Tumor necrosis factor-alpha-induced proliferation requires synthesis of granulocyte-macrophage colony-stimulating factor. AB - OBJECTIVE: Tumor necrosis factor- alpha (TNF-alpha) induces a variety of cellular responses, some of them being at least seemingly contradictory. Thus, we set out to find differences in the modes of proliferative and apoptotic responses to TNF- alpha. MATERIALS AND METHODS: We screened a panel of acute myeloid leukemia derived cell lines for TNF- alpha-responsiveness. In two lines (OCI-AML-1, OCI AML-11), TNF- alpha acted as an apoptotic agent; in others (HU-3, M-07e, TF-1), it had the opposite effect, preventing apoptosis and inducing proliferation. Direct and indirect signaling mechanisms, including NF-kappaB activation and cytokine synthesis, were analyzed. RESULTS: All cell lines tested expressed TNF- alpha receptors I and II and responded to TNF- alpha by upregulation of intercellular adhesion molecule-1. In contrast to granulocyte-macrophage colony stimulating factor (GM-CSF), TNF- alpha did not activate the MAP kinase and p70S6 kinase pathways. Nevertheless, inhibitors of these pathways clearly reduced the TNF-alpha-induced cell growth, indicating that TNF- alpha-proliferative cells produced a growth factor that induced proliferation upon stimulation of the above pathways. Anti-GM-CSF antibodies inhibited the TNF-alpha-induced growth, suggesting the presence of an autocrine loop for cell proliferation mediated by GM-CSF. Supporting this notion, TNF-alpha-induced upregulation of GM-CSF mRNA levels and protein secretion in the TNF-alpha-proliferative, but not in the TNF alpha-apoptotic cell lines. CONCLUSION: These data identify GM-CSF synthesis as an early and essential step in TNF- alpha-induced proliferation. We show for the first time that TNF-alpha-treated cell lines producing no or only minimal amounts of GM-CSF demonstrate an apoptotic phenotype, while cell lines with high GM-CSF expression rates can escape from growth arrest or even apoptosis. In this context, we discuss arguments pointing at NF-kappaB as regulator of GM-CSF synthesis and thus indirectly as regulator for the escape of TNF-alpha-induced apoptosis. PMID- 11008014 TI - Dynamic model of ex vivo granulocytic kinetics to examine the effects of oxygen tension, pH, and interleukin-3. AB - OBJECTIVE: Evaluating kinetics in hematopoietic cultures is complicated by the distribution of cells over various stages of differentiation and by the presence of cells from different lineages. Thus, an observed response is an integral response from distributed cell populations. Growth factors and other parameters can greatly affect the lineage and maturation stage of the culture outcome. To resolve the kinetics and more clearly define the differential effects of O(2) tension (pO(2)), pH, and interleukin-3 (IL-3) on granulopoiesis, a mathematical model-based approach was undertaken. MATERIALS AND METHODS: Granulocytic differentiation is described within a continuous, deterministic framework in which cells develop from primitive granulocytic progenitors to mature neutrophils. The model predicts two distributed populations-quiescent and cycling cells-by incorporating rates of growth, death, differentiation, and transition between quiescence and active cycling. The response of these four model processes to changes in the culture environment was examined. RESULTS: Model simulations of experimental data revealed the following: 1) pO(2) effects are exerted only on the growth rate but not maturation times. 2) pH effects between pH 7.25 and 7.4 on growth and differentiation are coupled; however, with increasing pH values, especially at pH 7. 6, the death rate for cells in the early stages of differentiation becomes increasingly significant. 3) The absence of IL-3 increases the death rate for primitive cells only minimally but markedly enhances the rate of differentiation through the myeloblast window in the differentiation pathway. The combined effects of these environmental factors can be predicted based on changes in the model parameters derived from the individual effects. CONCLUSIONS: Experimental data combined with mathematical modeling can elucidate the mechanisms underlying the regulation of granulopoiesis by pO(2), pH, and IL 3. The model also can be readily adapted to evaluate the effects of other culture conditions. The increased understanding of experimental results gained with this approach can be used to modify culture conditions to optimize ex vivo production of neutrophil precursors. PMID- 11008015 TI - Enhanced proteolysis of pre-mRNA splicing factors in myeloid cells. AB - OBJECTIVE: Molecular identification and characterization of the bone marrow nuclear protein detected by the B92 monoclonal antibody. MATERIALS AND METHODS: The protein was purified to homogeneity from acute myeloid leukemia cells and was subjected to peptide digestion and amino acid sequencing. Identified sequences were used to screen a bone marrow cDNA library in search of matching transcripts. The protein was further studied in different cells and tissues by examination of protease inhibitors and harsh lytic conditions and during apoptosis in HL-60 cells. RESULTS: We found that the apparent bone marrow specific protein is a 47 kD proteolytic cleavage product of PSF, an essential pre-mRNA splicing factor. PSF is completely cleaved to p47 during lysis of immature myeloid cells due to potent proteolytic activity found in these cells but is rare in other cells and tissues. Furthermore, p47 is abundant in intact normal and tumor myeloid cells while in other cell types it is undetectable. The cleavage of PSF is accompanied by digestion of the PTB splicing regulator but not other proteins tested. In contrast, during apoptosis PTB is degraded while PSF remains intact. CONCLUSIONS: The bone marrow 47 kD protein is a fragment constituting the N-terminal, protease resistant half of the splicing factor PSF. Proteolytic degradation of PSF specifically occurs in intact myeloid cells and this process is enhanced upon myeloid cell lysis. PMID- 11008016 TI - Quantitation of minimal residual disease in multiple myeloma using an allele specific real-time PCR assay. AB - OBJECTIVE: To develop a real-time PCR method, based on the 5'nuclease TaqMan technology, for quantitation of clonal cells in multiple myeloma (MM). MATERIALS AND METHODS: The real-time quantitative PCR method incorporates both an allele specific oligonucleotides (ASO) primer and an ASO dual-labeled fluorogenic probe (ASO TaqMan probe). The ASO primer and probe corresponded to the complementary determining region 3 (CDR3) of the rearranged immunoglobulin heavy chain gene (IgH). With the use of a sequence detector, PCR product accumulation was measured through the ASO TaqMan probe. The real-time PCR method was compared with flow cytometric quantitation of myeloma plasma cells. RESULTS: The application of the real-time quantitative ASO IgH PCR method is illustrated by a sequential analysis of minimal residual disease (MRD) in bone marrow (BM) samples from myeloma patients undergoing peripheral blood stem cell (PBSC) transplantation. The real time PCR method was able to quantitate residual malignant cells in BM samples from patients who were considered to be in complete remission. Further, it was illustrated that a potential problem in determining tumor cell content in myeloma BM samples is the heterogeneous infiltration of the marrow. CONCLUSION: The application of the real-time PCR method provides a sensitive, highly specific, and reproducible quantitation of myeloma cells. PMID- 11008017 TI - Transplantation and growth characteristics of human fetal lymph node in immunodeficient mice. AB - OBJECTIVE: The lymph node is an integral component of the immune system and the major site of antigen-dependent lymphocyte proliferation and differentiation. Development of animal models possessing functional primary human lymph nodes will have a significant impact on research in lymphopoiesis and immune response. To date, successful transplantation of primary human lymph nodes in rodents has not yet been reported. This work was undertaken to develop a reliable methodology to engraft primary human fetal lymph nodes in immunodeficient mice. MATERIALS AND METHODS: Three different sets of parameters, including three different transplantation sites in the mice, two different strains of immunodeficient mice, and two different preconditioning regimens, were evaluated. The growth characteristics of the implanted primary human fetal lymph nodes were examined 3 months after transplantation by histologic, immunocytochemical, and flow cytometric methods. RESULTS: Transplantation of primary human fetal lymph nodes into subcutaneous pouches in the ears in severe combined immunodeficiency (SCID) mice preconditioned with etoposide reproducibly give rise to >80% engraftment. The engrafted primary human fetal lymph nodes undergo massive growth (>200-fold) and retain the same histology and cellular composition as fresh human fetal lymph nodes from the same donors. CONCLUSIONS: We report, for the first time, the development of a reliable methodology to successfully engraft human fetal lymph node in SCID mice. The engrafted human lymph nodes are visible and accessible to experimental manipulations. This SCID-hu mouse model with human lymph node should provide a physiologically relevant system to investigate lymphopoiesis, immunologic response, and virus-mediated immunosuppression. PMID- 11008018 TI - Differences in megakaryocyte expansion potential between CD34(+) stem cells derived from cord blood, peripheral blood, and bone marrow from adults and children. AB - OBJECTIVE: Reinfusion of ex vivo expanded autologous megakaryocytes together with stem cell transplantation may be useful to prevent or reduce the period of chemotherapy-induced thrombocytopenia. We compared the megakaryocyte expansion potential of CD34(+) stem cells derived from different sources: cord blood (CB), peripheral blood (PB), bone marrow from adults (ABM), and bone marrow from children (ChBM). Three different growth factor combinations were tested to identify the best combination for each of the sources. MATERIALS AND METHODS: CD34(+) cells were isolated from CB, PB, ABM, or ChBM and cultured in an in vitro liquid culture system in the presence of thrombopoietin (Tpo), Tpo + interleukin (IL-1), or Tpo + IL-3. After 8 days, proliferation was determined and the cultured cells were identified with lineage-specific surface markers by flow cytometry. RESULTS: Cultures with ChBM-derived CD34(+) cells showed the lowest level of expansion of megakaryocytes and gave rise to more profound formation of myeloid and monocytic cells. In cultures with BM- or PB-derived cells, presence of IL-3 reduced the number of immature megakaryocytes (CD34(+)CD41(+) cells). However, in CB cultures, the number of CD34(+)CD41(+) cells was highest in cultures with Tpo + IL-3. Overall, cultures with CB CD34(+) cells yielded the highest number of megakaryocytes, but these cells showed reduced ploidization and lower level of CD41 expression, suggesting less maturation. CONCLUSIONS: Each of the different CD34(+) cell sources responded differently to cytokine stimulation. For PB and ABM, the cytokine combination Tpo + IL-1 is most suitable to obtain high numbers of both immature and mature megakaryocytes for transfusion purposes. For CB, Tpo + IL-3 is better. PMID- 11008019 TI - Effects of the protein tyrosine phosphatase CD45 on FcgammaRIIa signaling and neutrophil function. AB - OBJECTIVE: Neutrophil receptors for the Fc portion of IgG (FcgammaR) trigger immune responses following cross-linking by IgG-coated foreign particles or immune complexes. Membrane-associated CD45, a protein tyrosine phosphatase termed leukocyte common antigen, has been shown to be essential for antigen receptor kinase mediated signaling in lymphocytes, and we hypothesized that CD45 may play a similar role in FcgammaR-mediated signaling and immune function in human neutrophils. METHODS: The experimental approach was that of cell surface molecule ligation via cross-linking with specific antibodies. Antibody dependent cellular cytotoxicity (ADCC) was assessed using a single-cell plaque assay and IL-6 production measured using ELISA. Tyrosine phosphorylation levels were assessed with anti-phospho-tyrosine blots and F-actin polymerization by flow cytometry and confocal microscopy. RESULTS: Neutrophils pretreated with anti-CD45 had a reduced ability to perform ADCC compared to untreated neutrophils. FcgammaRIIa cross linking resulted in significantly increased concentrations of secreted IL-6 compared to untreated neutrophils, and IL-6 production was further enhanced by cocross-linking CD45 with FcgammaRIIa. Cross-linking CD45 alone also induced IL-6 production. FcgammaRIIa cross-linking resulted in increased protein tyrosine phosphorylation and F-actin polymerization in neutrophils. Cocross-linking CD45 with FcgammaRIIa resulted in abrogation of FcgammaRIIa mediated tyrosine phosphorylation and F-actin polymerization. CONCLUSIONS: These data provide evidence that CD45 can regulate or enhance the stimulation and function of human neutrophils mediated through FcgammaR(s). In addition, CD45 ligation may play an essential role in cytokine induction pathways that lead to inflammatory reactions in vivo. PMID- 11008020 TI - Kinetics of engraftment of CD34(-) and CD34(+) cells from mobilized blood differs from that of CD34(-) and CD34(+) cells from bone marrow. AB - OBJECTIVE: Mobilized peripheral blood (PB) progenitors are increasingly used in autologous and allogeneic transplantation. However, the short- and long-term engraftment potential of mobilized PB or bone marrow (BM) has not been directly compared. Although several studies showed that BM-derived Lin(-)CD34(-) cells contain hemopoietic progenitors, no studies have addressed whether Lin(-)CD34(-) cells from mobilized PB contain hemopoietic progenitors. Here, we compared the short- and long-term engraftment potential of CD34(+) cells and Lin(-)CD34(-) cells in BM and PB of normal donors who received 5 days of granulocyte colony stimulating factor (G-CSF). MATERIALS AND METHODS: 35 x 10(3) CD34(+) or Lin( )CD34(-) cells from G-CSF mobilized BM and PB of normal donors were transplanted in 60-day-old fetal sheep. Animals were evaluated 2 and 6 months after transplantation for human hemopoietic cells. In addition, cells recovered after 2 months from fetal sheep were serially passaged to secondary and tertiary recipients to assess long-term engrafting cells. RESULTS: Mobilized PB CD34(+) cells supported earlier development of human hemopoiesis than BM CD34(+) cells. When serially transferred to secondary and tertiary recipients, earlier exhaustion of human hematopoiesis was seen for PB than BM CD34(+) cells. A similar degree of chimerism was seen for Lin(-)CD34(-) cells from PB or BM in primary recipients. We again observed earlier exhaustion of human hemopoiesis with serial transplantation of PB than BM Lin(-)CD34(-) cells. CONCLUSIONS: Differences exist in the short- and long-term repopulating ability of cells in PB and BM from G-CSF mobilized normal donors, and this is independent of the phenotype. Studies are ongoing to examine if this reflects intrinsic differences in the repopulating potential between progenitors from PB and BM, or a lower frequency of long-term repopulating cells in PB than BM CD34(+) and Lin(-)CD34(-) cells, that may not be apparent if larger numbers of cells are transplanted. PMID- 11008021 TI - High frequency of long-term culture-initiating cells retain in vivo repopulation and self-renewal capacity. AB - OBJECTIVE: We wished to test if the long-term culture initiating cell (LTC-IC) assay measures primitive hematopoietic stem cells. An LTC-IC is defined by its ability to repopulate a stromal layer by forming colonies of myeloid cells. A negative well should never have received a stem cell, whereas a positive well should have been initiated by a stem cell. If these colonies were derived from stem cells, then a subset of the positive wells should retain stem cell activity. MATERIALS AND METHODS: Limiting dilution cultures were initiated on the stromal cell line S17. Individual clonal cultures from LTC-IC assays were assessed for repopulation capacity in W(14)W(41) mice. RESULTS: In long-term repopulation experiments, little activity was found in the negative wells, whereas 50% of the positive wells contained repopulating stem cells. The diverse in vivo repopulation patterns of the clonally derived stem cells suggest that this assay detects the full spectrum of stem cell types. Secondary transfers show that the clonally derived stem cells have self-renewal capacity. Experiments with mixtures of genetically distinguished cells showed that most (>90%) of the cultures were clonal. CONCLUSIONS: Our data present the first formal link between LTC-IC and repopulating stem cells. Moreover, the culture system presents a new way of generating a high frequency of clonally repopulating stem cells. PMID- 11008022 TI - Multi-lineage expansion potential of primitive hematopoietic progenitors: superiority of umbilical cord blood compared to mobilized peripheral blood. AB - OBJECTIVE: The majority of studies assessing ex-vivo expansion of primitive hematopoietic cells only address production of myeloid progeny whereas it may be more appropriate to maintain or expand progenitors that retain capacity for multilineage differentiation. In this study, we assessed the capacity of the murine fetal liver cell line AFT024 to expand primitive myeloid progenitors (LTC IC) and lymphoid progenitors (NK-IC) from umbilical cord blood (CB) and mobilized peripheral blood (PB) CD34(+)lin(-)38(-) cells. METHODS: Sorted cells were established in expansion cultures in direct contact with the feeder or in a transwell above the feeder (noncontact culture) and various combinations of Flt 3L (FL), stem cell factor, interleukin 7, thrombopoietin (Tpo), and macrophage inflammatory protein-1alpha added. Frequency of LTC-IC and NK-IC was assessed at day 0 and following 2 and 5 weeks expansion culture. RESULTS: CB contained significantly more LTC-IC and NK-IC at day 0 and showed an enhanced capacity for expansion compared to PB. The combination of FL and Tpo showed maximal expansion of CB LTC-IC and NK-IC at 5 weeks in both contact and noncontact conditions. In contrast, expansion of PB LTC-IC and NK-IC was maximal at 2 weeks and required multiple cytokines. CONCLUSIONS: These results demonstrate that AFT024 can expand primitive hematopoietic progenitors from CB and PB and expanded cells retain the capacity for myeloid and lymphoid differentiation. These findings emphasize the importance of assessing multi-lineage differentiation capacity following ex-vivo expansion. Elucidation of specific factors necessary for ex-vivo expansion will contribute to the development of a clinically applicable system. PMID- 11008023 TI - Second allogeneic stem cell transplantation using nonmyeloablative conditioning for patients who relapsed or developed secondary malignancies following autologous transplantation. AB - OBJECTIVE: Second allogeneic stem cell transplants for hematological malignancies are associated with a high incidence of transplant-related mortality due to the cumulative incidence of toxicity of the high-dose chemoradiotherapy traditionally used as an essential component of the conditioning. We have demonstrated previously that nonmyeloablative conditioning for primary allogeneic transplants from both sibling and unrelated donors results in minimal transplant-related toxicity and excellent stem cell engraftment. This study explores the possibility of using nonmyeloablative conditioning to minimize transplant-related toxicity in patients who have undergone second allogeneic transplants. PATIENTS AND METHODS: Twelve high-risk, heavily treated patients-five with acute myelogenous leukemia (AML); five with non-Hodgkin's lymphoma (NHL); one with Burkitt's lymphoma, and one with acute lymphoblastic leukemia (ALL)-underwent second allogeneic nonmyeloablative stem cell transplantation (NST) from human leukocyte antigen (HLA)-matched donors, 29 (median) (range 3-57) months following their first transplantation procedure. The conditioning consisted of fludarabine 30 mg/m(2) daily for 6 days, busulfan 4 mg/kg daily for 2 days, and anti-T-lymphocyte globulin 10 mg/kg daily for 4 days. Anti-graft-vs-host disease (anti-GVHD) prophylaxis consisted of cyclosporine A alone, 3 mg/kg. RESULTS: Engraftment was observed in all recipients, with complete and stable chimerism. None of the patients developed veno-occlusive disease of the liver or multi-organ failure. Five very high-risk patients with NHL (n = 3), Burkitt's lymphoma (n = 1), and AML (n = 1) relapsed 2 to 6 months post-transplant, and four of them died. Six patients appear to be disease-free after median follow-up of 23 months. One additional patient died from grade IV hemorrhagic cystitis. Actuarial survival and disease-free survival at 34 months are 56% and 50% respectively, with 95% confidence interval (25-78%). CONCLUSION: These results suggest that nonmyeloablative conditioning significantly reduces transplant-related toxicity, thus making a second transplant feasible. PMID- 11008024 TI - Refractive surgery for keratoconus: a new approach. PMID- 11008025 TI - Consultation section: refractive surgical problem. PMID- 11008026 TI - Refractive surgical problem PMID- 11008027 TI - Refractive surgical problem PMID- 11008028 TI - Refractive surgical problem PMID- 11008029 TI - Refractive surgical problem PMID- 11008030 TI - Refractive surgical problem PMID- 11008031 TI - Refractive surgical problem PMID- 11008032 TI - Refractive surgical problem PMID- 11008033 TI - Refractive surgical problem PMID- 11008034 TI - Refractive surgical problem PMID- 11008035 TI - Refractive surgical problem PMID- 11008036 TI - Cataract in enucleated goat eyes: training model for phacoemulsification. AB - We developed an inexpensive, reproducible technique of inducing cataract in enucleated animal eyes for phacoemulsification training. Injection of 0.3 to 0.5 mL of formalin 20% in the central and paracentral portion of the crystalline lens in a goat's eye induced cataract in about 5 to 10 minutes. Varying the interval would enable novice surgeons to practice different techniques of phacoemulsification based on cataract hardness. PMID- 11008037 TI - Correcting keratoconus with intracorneal rings. AB - PURPOSE: To evaluate the potential of intrastromal corneal ring technology (Intacs, KeraVision) to correct keratoconus without central corneal scarring. SETTING: Department of Ophthalmology, Brest University Hospital, Brest, France. METHODS: In this prospective, noncomparative, interventional case series, Intacs segments were implanted in 10 keratoconic eyes with clear central corneas and contact lens intolerance after corneal pachymetry was checked. Segment thicknesses varied based on corneal topography analysis. RESULTS: No intraoperative complications occurred. The mean follow-up was 10.6 months. Postoperative results revealed a reduction in astigmatism and spherical correction and an increase in topographical regularity and increased uncorrected visual acuity. CONCLUSION: Intacs technology can reduce the corneal steepening and astigmatism associated with keratoconus. PMID- 11008038 TI - Theoretical analysis of ablation depths and profiles in laser in situ keratomileusis for compound hyperopic and mixed astigmatism. AB - PURPOSE: To compare the theoretical ablation depths and profiles of 4 treatment strategies for compound hyperopic and mixed astigmatism. SETTING: Theoretical analysis. METHODS: Corneal contour drawings of theoretical corneal ablation profiles during laser in situ keratomileusis (LASIK) and photoastigmatic refractive keratectomy were made. The depths of tissue ablation in 4 treatment strategies for compound hyperopic astigmatism (Groups 1 to 4) and for mixed astigmatism (Groups 5 to 8) were compared: (1) combined hyperopic spherical and myopic cylindrical treatments (Groups 1 and 5); (2) combined spherical and hyperopic cylindrical treatments (Groups 2 and 6); (3) combined cylindrical treatments (Groups 3 and 7); (4) combined cross-cylinder and spherical equivalent treatments (Groups 4 and 8). RESULTS: In compound hyperopic astigmatism, the 4 approaches resulted in identical final curvatures, but the ablation depths were greatest in Group 1 (combined hyperopic spherical and myopic cylindrical treatments). The smallest amount of ablation occurred in Group 2 (combined hyperopic spherical and hyperopic cylindrical treatments) and Group 3 (combined hyperopic cylindrical treatments), which had similar tissue ablation patterns. In mixed astigmatism, the greatest ablation depth was in Group 5, followed by Group 8, and Groups 6 and 7. The tissue ablation depths and profiles were similar in Groups 6 and 7. CONCLUSION: The treatment approaches in Groups 2, 3, 6, and 7 (which avoided the use of minus cylinder) resulted in the smallest degree of stromal ablation. Patients with compound hyperopic or mixed astigmatism may benefit from reduced ablation depths by deferring treatment until hyperopic cylindrical and/or combined cylindrical treatments are available. PMID- 11008039 TI - Comparison of optical zones in hyperopic laser in situ keratomileusis: 5.9 mm versus smaller optical zones. AB - PURPOSE: To compare the results of hyperopic laser in situ keratomileusis (LASIK) with a 5.9 mm optical zone (OZ) with those with smaller zones (4.4 to 5.5 mm). SETTING: Instituto de la Vision, Buenos Aires, Argentina. METHODS: The results of LASIK with a 5.9 mm OZ (147 cases) were compared with those in a previously reported group treated with OZs of 4.4 to 5.5 mm (679 cases). In the 5.9 mm group, 31.3% (46 eyes) had low hyperopia, 46.9% (69 eyes) had moderate hyperopia, and 21.8% (32 eyes) had high hyperopia. In the smaller OZ group, follow-up was 1 month in 79.4% (539 eyes), 3 months in 75.5% (501 eyes), 6 months in 68.5% (465 eyes), and 1 year in 38.3% (260 eyes). The hyperopic population studied was divided into 3 subgroups based on the preoperative spherical equivalents of the manifest refraction: subgroup A, low hyperopia: < or = +2.0 diopters (D); subgroup B, moderate hyperopia: +2.0 to +3.0 D; and subgroup C, high hyperopia: > +3.0 D. The following parameters were measured postoperatively: uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refraction (evolution and distribution), and visual acuity lines gained and lost. RESULTS: The preoperative mean spherical equivalents in the 5.9 mm OZ group were +1.47 +/- 0.41 (SD) (subgroup A); +2.98 +/- 0.41 (subgroup B); and +5.13 +/- 0.61 (subgroup C). In the smaller OZ group, they were +1.31 +/- 0.74, +2.56 +/- 0.28, and +5.28 +/- 0. 69, respectively. At 12 months, the distribution of eyes in the 5.9 mm OZ group with refractions within +/-1.0 D were as follows: subgroup A, 100%; subgroup B, 100%; and subgroup C, 94.2%. In the smaller OZ group, the distributions were 100%, 95.3%, and 71.4%. In the 5.9 mm and the smaller OZ groups, the UCVA was 20/40 or better in 92.0% and 81.8% of eyes, respectively, in subgroup A; 94.6% and 100%, respectively, in subgroup B; and 76.5% and 77.9%, respectively, in subgroup C. The percentage of eyes with 0 +/- 1 line of BCVA 12 months after the procedure was also determined in the 5.9 mm OZ group and compared with the percentages in the smaller OZ group. CONCLUSION: Results of hyperopic LASIK with a 5.9 mm OZ in eyes with low, moderate, and high hyperopia are more stable, predictable, and safe than those in eyes with OZs smaller than 5.9 mm. PMID- 11008040 TI - Accuracy and predictability of intraocular lens power calculation after photorefractive keratectomy. AB - PURPOSE: To investigate the accuracy and predictability of intraocular lens (IOL) power calculation in postoperative photorefractive keratectomy (PRK) eyes. SETTING: Gimbel Eye Centre, Calgary, Alberta, Canada. METHODS: The results in 5 cataract surgery eyes that had had PRK were analyzed retrospectively. Target refractions based on actual and refraction-derived keratometric values were compared with postoperative achieved refractions. The target refractions calculated using 5 IOL formulas and 2 A-constants were also compared with the achieved refractions. RESULTS: In postoperative PRK eyes, the power calculation was more accurate and predictable when the smaller of either the actual or refraction-derived keratometric value was used to calculate the IOL power. The difference between target and achieved refractions appeared smaller when the Binkhorst formula was used. No significant hyperopic shift was observed after cataract surgery. CONCLUSION: The smaller of the actual or the refraction-derived keratometric value is recommended for calculating IOL power in post-PRK eyes. PMID- 11008041 TI - Laser in situ keratomileusis to correct residual myopia and astigmatism after radial keratotomy. AB - PURPOSE: To evaluate the safety and efficacy of laser in situ keratomileusis (LASIK) in selected post-radial-keratotomy (RK) eyes with residual myopia and astigmatism. SETTING: TLC-The Brea Laser Eye Center, Brea, California, USA. METHODS: Nine eyes of 6 patients who had had RK but had residual myopia and/or astigmatism had LASIK. All RK eyes had 8 radial incisions, were more than 1 year post-RK, had no epithelial inclusion cysts or corneal disease, and had had no subsequent ocular surgery. Follow-up was a minimum of 13 months, at which time uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), manifest refraction, cycloplegic refraction, keratometry, central and peripheral pachymetries, intraocular pressure, and a subjective assessment of visual function were obtained. RESULTS: At the last follow-up, the mean spherical equivalent (SE) was -0.156 diopter (D) +/- 0.174 (SD). All eyes treated for distance vision had a UCVA of 20/25 or better. No patient lost BCVA. No intraoperative or postoperative complications occurred. Seven eyes had morning and evening measurements. The mean change in manifest SE from morning to evening was -0.143 D. Six of the 7 eyes (86%) had 0 to 1 Snellen line change in UCVA from morning to evening. The subjective questionnaire revealed a high degree of satisfaction with overall vision, minimal glare, and less fluctuation in daily vision than before LASIK. CONCLUSION: Laser in situ keratomileusis is safe and efficacious for reducing residual myopia and astigmatism in properly selected RK patients. PMID- 11008042 TI - Outcome of flap subluxation after laser in situ keratomileusis: results of 6 month follow-up. AB - PURPOSE: To evaluate the long-term outcome in cases with flap subluxation after laser in situ keratomileusis (LASIK). SETTING: Private clinic in Turkey. METHODS: Data of all patients who had LASIK were documented and the follow-ups of those who had flap subluxation evaluated. The number of LASIK cases was 1481 and the number of flap subluxations, 21. Laser in situ keratomileusis was performed with the Moria microkeratome and the VISX Twenty/Twenty B excimer laser using a multizone profile and the sutureless hinged corneal flap technique. RESULTS: The incidence of flap subluxation was 1.42%. Statistical analysis of uncorrected visual acuities, best corrected visual acuities, and mean spherical equivalents in eyes with and without flap subluxation was performed. No significant differences were found, although the results in eyes with flap subluxation were worse than those in eyes without flap subluxation. Additional complications were also higher in eyes with flap subluxation. CONCLUSION: Flap subluxation is a serious but manageable complication of LASIK. If it is detected early, the visual prognosis is as good as that in eyes without flap subluxation. PMID- 11008043 TI - Multicenter study of the Artisan phakic intraocular lens. AB - PURPOSE: To assess the Artisan intraocular lens to correct myopia in phakic eyes. SETTING: European multicenter study sponsored by Ophtec BV, Groningen, The Netherlands. METHODS: In this prospective multicenter clinical study, the Artisan lens was implanted in 518 eyes between September 1991 and October 1999. The power of the lenses ranged from -5.0 to -20.0 diopters (D). Follow-up examinations were performed at 6 months and 1, 2, and 3 years. Follow-up ranged from 6 months (n = 454) to 3 years (n = 249). The preoperative uncorrected visual acuity (UCVA) was not recorded but was estimated to be worse than 0.1. The preoperative mean best spectacle-corrected visual acuity (BSCVA) was 0.67 +/- 0.26 (SD). Endothelial cell counts were done at 6 months and 1, 2, and 3 years in a subgroup of 129 eyes. RESULTS: A UCVA of 20/40 or better was observed in 76.8% of eyes regardless of the postoperative goal. A BSCVA of 20/40 or better was observed in 93.9% of eyes and remained stable throughout the follow-up. Of the eyes with extremely high myopia (>-15.0 D), 63.3% gained 2 or more lines of BSCVA; of those with moderate myopia (-5.0 to -10.0 D), 23.5% gained 2 or more lines. The mean endothelial cell density change was 4.8% at 6 months, 2.4% at 1 year, 1.7% at 2 years, and 0.7% at 3 years. The incidence of persistent adverse events at 3 years was relatively low. Secondary surgical interventions included repositioning of the lens because of poor initial placement and lens exchange because of preoperative power calculation errors. Glare and halo effects during night driving were noted and were related to large pupils in young patients. CONCLUSION: The Artisan lens is a safe, stable, efficacious, and predictable method to correct -5.0 to -20.0 D of myopia. This study suggests that the corneal endothelial cell loss is stabilized to the physiologically normal level after 3 years. PMID- 11008044 TI - Long-term results of the foldable CeeOn Edge intraocular lens. AB - PURPOSE: To determine the long-term efficacy and safety of the foldable silicone CeeOn Edge(R) model 911 intraocular lens (IOL). SETTING: Eye Clinic, Klinikum Minden, Germany. METHODS: A high-refractive-index silicone lens with a sharp edge was implanted after phacoemulsification in 42 eyes of 36 patients. Best corrected visual acuity (BCVA) and occurrence of complications were evaluated during a 3 year follow-up. RESULTS: No significant postoperative complications were seen. At 3 years, 26 eyes of 22 patients were available for examination. The BCVA was 0.5 (20/40) or better in 92% of eyes and 1.0 (20/20) or better in 39%. No posterior capsule opacification (PCO) was observed, and neodymium:YAG capsulotomy was not indicated in any patient. CONCLUSIONS: This pilot study indicates that the performance and safety of the foldable silicone CeeOn Edge lens are excellent. The results support the theory that a sharp edge on the IOL prevents the development of PCO. PMID- 11008045 TI - Posterior capsule opacification 3 years after implantation of an AcrySof and a MemoryLens in fellow eyes. AB - PURPOSE: To compare the rates of lens epithelial cell (LEC) migration and posterior capsule opacification (PCO) 1 and 3 years after sutureless small incision phacoemulsification and in-the-bag implantation of 2 acrylic polymer intraocular lenses (IOLs)-the AcrySof and MemoryLens-in fellow eyes of patients. SETTING: Eye Clinic, Beyoglu Education and Research Hospital, Istanbul, Turkey. METHODS: Fifty patients with no systemic or ocular problems that would interfere with postoperative visual acuity were included in this prospective study. Each patient had in-the-bag implantation of an AcrySof IOL in 1 eye and a MemoryLens in the fellow eye in a randomized fashion after uneventful phacoemulsification through a sutureless clear corneal incision. RESULTS: At 1 year (n = 32 patients), there was no significant difference between fellow eyes in postoperative best corrected visual acuity (BCVA) and contrast sensitivity. In the MemoryLens group, 10 eyes (31.3%) had PCO and 9 (28.1%), LEC migration. In the AcrySof group, no eye had PCO and 2 eyes (6.3%) had LEC migration (P <.001). At 3 years (n = 21 patients), 1 eye (4.7%) in the AcrySof group had PCO and 3 eyes (14.4%) had LEC migration without PCO. In the MemoryLens group, 1 eye (4.7%) had a clear posterior capsule, 11 eyes (52.4%) had LEC migration, and 9 eyes (42.9%) had PCO (P <.001). A neodymium:YAG capsulotomy was required in 4 eyes (19.0%) in the MemoryLens group but no eye in the AcrySof group. At 3 years, BCVA was lower in the MemoryLens group than in the AcrySof group (P <.05). CONCLUSION: The 3 year clinical data of fellow eyes indicate that the AcrySof IOL causes less PCO than the MemoryLens. PMID- 11008046 TI - Influence of laser posterior capsulotomy on anterior chamber depth, refraction, and intraocular pressure. AB - PURPOSE: To evaluate the effect of neodymium:YAG (Nd:YAG) laser posterior capsulotomy on anterior chamber depth (ACD), intraocular pressure (IOP), and refraction, including spherical equivalent (SE) and various forms of astigmatism. SETTING: Taipei Municipal Yang-Ming Hospital, Taipei, Taiwan. METHODS: Fifty three eyes of 46 patients with posterior capsule opacification were included in this prospective study. An Nd:YAG laser was used to perform posterior capsulotomy. Patients' ACD, IOP, and refraction were measured before the capsulotomy and 30 minutes, 1 week, and 1 and 3 months after. RESULTS: There were no statistically significant differences in ACD, IOP, or SE (P =.201, P =.465, and P =.109, respectively). However, there were significant decreases in the magnitudes of refractive astigmatism and residual astigmatism after laser treatment (P =.012 and P <.001, respectively). CONCLUSIONS: Although an Nd:YAG laser posterior capsulotomy did not significantly change ACD, IOP, or SE, it decreased the magnitudes of refractive astigmatism and residual astigmatism 1 week postoperatively. These stabilized thereafter. PMID- 11008047 TI - Retinal breaks before and after neodymium:YAG posterior capsulotomy. AB - PURPOSE: To determine the frequency of asymptomatic retinal breaks before and after neodymium:YAG (Nd:YAG) laser posterior capsulotomy. SETTING: Department of Ophthalmology, Helsinki University Central Hospital, Helsinki, Finland. METHODS: Of 350 consecutive patients referred for their first laser Nd:YAG posterior capsulotomy, 235 eligible eyes were enrolled and 220 eyes completed the study. A vitreoretinal surgeon looked for retinal breaks using binocular indirect ophthalmoscopy with scleral indentation. The eyes were examined 1 week before and 1 hour and 1 month after the posterior capsulotomy. The number, type, and location of retinal breaks were recorded. RESULTS: The median age of eligible patients who did not participate in the study was higher than that of enrolled patients (79.6 versus 74.4 years; P =.0005). The mean axial length, median time from cataract surgery, and the course of cataract surgery were comparable in both groups. Before the posterior capsulotomy, an untreated retinal break was diagnosed in 4 of the 235 eyes (1.7%; 95% confidence interval [CI] 0 to 4) scheduled for surgery and an undiagnosed retinal detachment was present in 2 additional eyes (0.9%; 95% CI 0 to 3). An asymptomatic retinal break was also present in 4 fellow eyes (1.7%, 95% CI 0 to 4). No new breaks developed during Nd:YAG posterior capsulotomy using a median total energy of 51 mJ (range 10 to 901 mJ) and a median number of 22 applications (range 4 to 341 applications) and resulting in an opening with a median largest diameter of 3.4 mm (range 2.0 to 4.6 mm). In 1 treated eye (0.4%; 95% CI 0 to 2), a new retinal break had developed by 1 month postoperatively. CONCLUSIONS: The observed 2.1% frequency of asymptomatic retinal breaks that had escaped the attention of the referring ophthalmologist or had developed by 1 month after Nd:YAG posterior capsulotomy can be contrasted with the 0.5% to 2.0% frequency of retinal detachment reported in the literature. However, it is not known which proportion of such asymptomatic breaks, if any, will progress to detachment after Nd:YAG laser posterior capsulotomy. PMID- 11008048 TI - Diplopia after cataract surgery. AB - PURPOSE: To evaluate the cause of diplopia after cataract surgery. SETTING: Cataract surgery at 7 hospitals and examination of diplopia at a central eye hospital. METHODS: This study comprised 18 eyes of 17 patients with diplopia that developed after cataract surgery in which retrobulbar anesthesia was used. The Hess screen test was done to diagnose oculomotor dysfunction. RESULTS: Several cases showed superior or inferior deviation of the globe, but most patients had nonuniform disturbances of eye movement. Examination of 3 patients by the Hess chart within 1 week after surgery showed paralysis of eye muscles but an overaction at a later stage, evident by reversal of eye position 1 month later. Surgery for strabismus was performed in 6 cases. One case with diplopia improved spontaneously 3 months after cataract surgery and achieved good alignment. CONCLUSIONS: The Hess screen test was useful for comparing changes in oculomotor function before and after surgery. Oculomotor dysfunction after cataract surgery may be caused directly by traumatic injury during administration of anesthesia or surgery using bridle sutures or indirectly from sensitivity to anesthetic agents. PMID- 11008049 TI - Incidence of ocular misalignment and diplopia after uneventful cataract surgery. AB - PURPOSE: To evaluate the incidence of ocular misalignment and diplopia after uneventful cataract surgery. SETTING: An outpatient private practice eye institute. METHODS: One hundred thirty-eight patients referred to 1 cataract surgeon were prospectively evaluated. Orthoptic evaluations were performed within 1 month before and then 1 day, 1 week, and 1 month after cataract surgery. Anesthesia was by retrobulbar injection, and cataract extraction was done by phacoemulsification. RESULTS: Cataract surgery was performed in 118 patients. Preoperatively, 16 patients had ocular misalignment; 10 were phoric, 4 were intermittently tropic, and 2 were tropic. Follow-up evaluation was obtained for 101 patients (86%) at 1 day, 91 (77%) at 1 week, and 88 (75%) at 1 month. A change in ocular alignment occurred in 22 of 101 patients (22%) at 1 day, 9 of 91 (10%) at 1 week, and 6 of 88 (7%) at 1 month. Only 1 patient who had a change in alignment at 1 month was symptomatic. CONCLUSIONS: A persistent change in ocular alignment after uneventful cataract surgery occurred in 7% of patients. However, symptomatic diplopia was uncommon (1 in 118; 0.85%) in this relatively small series. PMID- 11008050 TI - Phacoemulsification in eyes with a small pupil. AB - PURPOSE: To evaluate the intraoperative difficulties and long-term outcome of phacoemulsification through a small pupil using minimal iris manipulation. SETTINGS: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. METHODS: This prospective study included 30 consecutive eyes with a maximally dilated pupil size of 4.0 mm or smaller. Except synechiolysis and occasional pupil retraction with a chopper, no iris manipulation was performed. The phacoemulsification technique included creation of deep central space, use of a low aspiration flow rate with appropriate vacuum, and step-by-step chop in situ and lateral separation of the nucleus. Patients were examined 1 day, 1 week, 1 and 3 months, and 1 year postoperatively. Specular microscopy was performed at 1 month and 1 year. RESULTS: Fourteen eyes had chronic iritis, 12 had glaucoma surgery, 2 had pseudoexfoliation, and 2 had a rigid pupil. Mean follow-up was 13.6 months. Eighteen eyes (60%) had hard cataract. Mean preoperative pupil size was 2.80 mm +/- 0.75 (SD). Mean pupil size before the capsulorhexis was initiated was 4.42 +/ 0.58 mm. The iris was bruised in 10 eyes during cortex removal. Trace amounts of cortex were left in 4 cases. Two eyes had increased intraocular pressure in the early postoperative period. Six patients had a minimal amount of retained viscoelastic material (pseudofibrin), and 2 patients developed a sterile hypopyon. Sixteen eyes had cell deposits on the IOL surface and 12 eyes, fine uveal pigments. In 2 eyes with previous glaucoma surgery and in all eyes with iritis except 2, posterior synechias reformed. CONCLUSIONS: Successful phacoemulsification was done with minimal or no pupil-widening maneuvers, restoring the preoperative pupil configuration. PMID- 11008051 TI - Surgical management of congenital cataract associated with severe microphthalmos. AB - PURPOSE: To report the results of cataract surgery in children with severe microphthalmos and congenital cataract. SETTING: Seoul National University, Department of Ophthalmology, Seoul, Korea. METHODS: Retrospective studies were conducted by reviewing the charts of 20 eyes of 11 patients with severe microcornea and microphthalmos or severe microcornea with a corneal diameter smaller than 9.0 mm. The patient pool was divided according to surgical method, specifically by incision site: an anterior group that had corneal limbal incisions and a posterior group treated via pars plicata incisions. Preoperative examinations included bilaterality, corneal diameter, axial length, and eye and systemic abnormalities. Postoperative results and complications of the 2 operative methods were compared. RESULTS: All 11 patients had bilateral congenital cataract and microphthalmos. Except in 1 case, surgeries were done in patients ranging in age from 2 to 16 months. Mean follow-up was 2 years. Corneal diameter was smaller than 9.0 mm in all 20 eyes and smaller than 7.5 mm in 11 eyes. Ten eyes were categorized into the anterior group, and the other 10 eyes were placed in the posterior group. Postoperative complications included secondary membranes in 3 eyes in the posterior group and corneal opacity in 3 eyes in the anterior group. CONCLUSION: The surgical management of children with congenital cataract and severe microphthalmos is recommended as a way to improve vision but must be performed carefully to avoid complications. PMID- 11008052 TI - Acular as a single agent for use as an antimiotic and anti-inflammatory in cataract surgery. AB - PURPOSE: To assess the safety and effectiveness of ketorolac tromethamine 0. 5% (Acular) as a cost-efficient single agent to prevent intraoperative miosis and postoperative inflammation in cataract surgery. METHODS: Both eyes of 26 patients were randomized to receive Acular preoperatively and postoperatively or flurbiprofen sodium (Ocufen) preoperatively and prednisolone acetate 1% (Pred Forte) postoperatively. Time scheduled between procedures was from 2 weeks to 1 month. Pupil dilation was measured preoperatively, intraoperatively, and at the end of surgery. Cell and flare were measured 1 day, 1 week, and 1 month postoperatively. RESULTS: A comparison of the Acular and the Ocufen/Pred Forte groups (n=22) showed no statistically significant differences in dilation (preoperative versus postpostoperative) or cell and flare postoperatively. CONCLUSIONS: Using Acular as a single agent was as effective as the combination of preoperative Ocufen and postoperative Pred Forte in preventing intraoperative miosis and postoperative inflammation in cataract surgery. The use of Acular as a single agent could save the expense of using separate anti-inflammatory and antimiotic preparations preoperatively and postoperatively, enhancing convenience for the surgeon and surgical facility. PMID- 11008053 TI - Gentian violet solution for staining the anterior capsule. AB - PURPOSE: To evaluate the histopathological changes after injecting gentian violet solution into the anterior chamber of rats and to describe a technique that uses gentian violet to allow a clear view of the anterior capsule during continuous curvilinear capsulorhexis (CCC) in human eyes with white mature cataract. SETTING: Department of Ophthalmology, University of Dicle, Diyarbakir, Turkey. METHODS: In this masked, experimental study (first stage), 0.05 mL of gentian violet 0.01% or 0.001% solution or balanced salt solution (BSS) (control group) was injected into the anterior chamber of 30 eyes of 30 Wistar albino rats. One, 24, and 48 hours after injection, 4 eyes in each group and 2 eyes in the control group were enucleated, and histopathological examination was performed. In the second stage, these solutions were used for staining the anterior capsule in the 18 human eyes with white mature cataract. The success rate of CCC and intraoperative and postoperative complications were evaluated. RESULTS: Histopathological examination revealed no pathology in any group. A CCC was completed in all cases. No intraoperative or postoperative complications were observed in human eyes except mild corneal edema and mild inflammatory reaction in the anterior chamber that improved within 1 week. Mean follow-up was 3.4 months. Visualization of the anterior capsule was better with gentian violet 0.01% solution. CONCLUSIONS: Gentian violet solutions at 0.01% and 0.001% concentrations had no evident toxic effect that caused significant histopathological changes. The staining technique was practical and helped the surgeon visualize the anterior capsule. However, gentian violet may have adverse effects that lead to corneal edema. PMID- 11008054 TI - Clinical results using the Holladay 2 intraocular lens power formula. AB - PURPOSE: To analyze the accuracy of the Holladay 2 formula, which has been proposed as an improvement over the original Holladay formula. METHODS: This retrospective analysis comprised 317 eyes operated on by 1 surgeon using 1 technique and 1 intraocular lens style in a specialty practice. Because the Holladay 2 formula has yet to be published, its accuracy can only be analyzed using the commercially available Holladay IOL Consultant computer program to compare it to the Holladay 1, Hoffer Q, and SRK/T formulas. Defined axial length ranges were analyzed individually. RESULTS: A lower mean absolute error (MAE) trend was found for the average length eye (22.0 to 24.5 mm) by the Holladay 1 and Hoffer Q formulas. For short eyes (< 22.0 mm), the Hoffer Q and Holladay 2 perform better. The SRK/T consistently showed a trend toward the lowest MAE in all long eyes (>24.5 mm) as well as the subdivisions of medium long (24.5 to 26.0 mm) and very long (>26.0 mm). The Holladay 2 trended toward the least accurate (MAE) of the 4 formulas in all ranges of axial length except the shortest and the very longest. It appears to perform poorer in average and medium long eyes. CONCLUSIONS: Although the Holladay 2 formula has improved its MAE accuracy in short eyes, it was not more accurate than the Hoffer Q. The changes made in the formula to effect this improvement in MAE seem to have sacrificed the accuracy of the original Holladay formula in eyes with average and medium long axial lengths. PMID- 11008055 TI - Potential acuity meter accuracy in cataract patients. AB - PURPOSE: To determine potential acuity meter (PAM) accuracy in patients with cataract as the only ocular disease. SETTING: Hospital de Clinicas de Porto Alegre, Brazil. METHODS: This prospective study comprised 74 eyes with cataract. Patients with other ocular pathology diagnosed before or after surgery, as well as those with complicated surgeries, were excluded. All patients had PAM evaluation before surgery; these results were compared to the final postoperative visual acuity. Patients were divided into 4 cataract groups based on the Lens Opacity Classification System II: Group 1, early cataracts (nuclear opalescence [NO] 1 and/or posterior subcapsular cataracts [P] 1 and/or cortical opacity [C] < 2); Group 2, definite cataracts (NO2 and/or C2, with or without P1); Group 3, posterior cataracts (P2 or P3); Group 4, mature cataracts (N3 and/or C > or = 3, with or without P1). The PAM result was considered accurate if it was within 2 lines of the postoperative best spectacle-corrected visual acuity (BSCVA) 3 months after surgery. RESULTS: All patients had a BSCVA of 20/20. Accuracy of the PAM was 58.3% in Group 1, 50.0% in Group 2, 27.8% in Group 3, and 6.7% in Group 4 (P =.000 017). Patient age did not influence the results. CONCLUSIONS: The PAM may be used as a complementary tool in evaluating early and nondense cataracts; however, patients should not be excluded as candidates for cataract surgery based solely on PAM results. PMID- 11008056 TI - Osmolality of various viscoelastic substances: comparative study. AB - PURPOSE: To determine the osmolality of various viscoelastic substances. SETTING: Department of Ophthalmology, Johannes Gutenberg-University, Mainz, and Institute for Medical Device Testing, Memmingen, Germany. METHODS: The analysis was carried out according to the European Pharmacopoeia by means of a calibrated osmometer using the freezing-point depression method. Each syringe was analyzed as a duplicate analysis. RESULTS: Mean osmolalities (mOsmol/kg) of the sodium hyaluronate viscoelastic substances were Allervisc 299; Allervisc Plus 307; Amvisc Plus 335; AMO Vitrax 284; Biolon 279; Dispasan 311; Dispasan Plus 314; Healon 295; Healon GV 312; Healon5 322; HYA-Ophtal 376; Microvisc 313; Microvisc Plus 341; Provisc 307; Rayvisc 312; Viscoat 340; Visko 296; Visko Plus 319. Mean osmolarities of the hydroxypropyl methylcellulose viscoeslatic substances were Adatocel 278; HPMC Ophtal L 358; HPMC Ophtal H 360; La Gel 317; OcuCoat 309; Visco Shield 376. CONCLUSIONS: There were significant differences in osmolality among viscoelastic substances, which may explain the differences in corneal thickness after cataract surgery. A viscoelastic substance with an osmolality of 305 mOsmol/kg or slightly higher is preferable, especially in patients with a compromised corneal endothelium. PMID- 11008057 TI - Intraocular pressure development in the rabbit eye after aqueous exchange with ophthalmic viscosurgical devices. AB - PURPOSE: To evaluate an animal model used to study intraocular pressure (IOP) development after aqueous exchange with several commercially available ophthalmic viscosurgical devices (OVDs). SETTING: Department of Preclinical Ophthalmology, Pharmacia & Upjohn, Uppsala, Sweden. METHODS: Albino rabbits (New Zealand White) were used. Aqueous humor (50 microL) was exchanged with 8 OVDs. The IOP was measured every second hour for 12 hours and then 24, 48, and 72 hours after aqueous exchange using a pneumotonometer (Modular One, Bio-Rad Digilab Inc). A minimum of 7 eyes was used for each OVD. Healon was used as control in 1 eye in all experiments. The OVDs were Viscoat (chondroitin sulfate-sodium hyaluronate), Provisc (sodium hyaluronate), Biolon (sodium hyaluronate), Healon GV, Healon5, Ophthalin (sodium hyaluronate), Ocucoat (hydroxypropyl methylcellulose), and Ivisc (sodium hyaluronate). RESULTS: All OVDs caused a postoperative increase in IOP. At 24 hours, the IOP was at the preoperative levels. However, there was considerable variation in the maximum IOP value and when this value occurred. The maximum value appeared to depend on the concentration of the rheologically active substance in the product and the time of the average molecular mass. There was also an increase in central corneal thickness, with a maximum increase 24 hours after the exchange but a large variation among animals. CONCLUSION: The animal model appears to be useful for comparing various OVDs, and the results may serve as a guide for the design of clinical studies of new products. PMID- 11008058 TI - Arteriovenous fistula induced by a peribulbar nerve block. AB - Arteriovenous fistula (AVF) of the head and neck region is an uncommon clinical condition that can be of congenital or acquired etiology. We report a case of AVF of the left supraorbital vessels that developed after a peribulbar nerve block was given for cataract surgery. PMID- 11008059 TI - Black intraocular lens for leukocoria. AB - We describe a 33-year-old man with leukocoria and no visual potential who obtained an excellent cosmetic result after cataract extraction and implantation of an intraocular lens with an entirely black optic. PMID- 11008060 TI - Phacoemulsification in a patient with marked cervical kyphosis. AB - A patient with long-standing ankylosing spondylitis and chronic uveitis needed cataract extraction in his only eye. Extensive spinal deformities, including cervical kyphosis, prevented him from being positioned satisfactorily for surgery using a routine head-end or temporal position for the surgeon. The best possible position for surgery was achieved using an orthopedic operating table, which allowed the patient's head to be reclined to a position of 60 degrees to the horizontal. Successful combined phacoemulsification and trabeculectomy was then performed, although the angle of approach for the surgeon and the operating microscope was awkward. PMID- 11008061 TI - Dimerization of G-protein-coupled receptors: implications for drug design and signaling. PMID- 11008062 TI - Dimerization of G-protein coupled receptors. PMID- 11008063 TI - Opioids and their complicated receptor complexes. AB - No field more eagerly awaits a molecular clarification for G-protein coupled receptor (GPCR) dimerization than the opioid receptor field. Extensive evidence of pharmacological and functional interactions between opioid receptor types has primed this field for such a resolution. In retrospect, much of the data collected on synergy between different opioid receptor types may represent the functional correlate for the newly found opioid receptor dimerization. While previous reports of functional synergy have been, for the most part, consistent in demonstrating cross-regulation between two receptor types, the lack of highly receptor-selective ligands allowed skeptics to remain doubtful over the interpretations of these results. Today, two important developments in the opioid receptor field help reinvigorate the hypothesis of functional, cross-modulating opioid receptor complexes: (1) The existence of highly selective ligands which eliminate any possibility of cross-reactivity between receptor types, and (2) the discovery that opioid receptors and a number of other GPCRs exist as dimers in biochemical, functional and pharmacological assays. It is with these new tools that we seek to understand the mechanisms and implications of dimerization. Initial results of these studies have demonstrated that the dimerization of opioid receptors may help consolidate several pharmacological findings that have remained unanswered. In this review we present biochemical, pharmacological and functional evidence for opioid receptor complexes and add evidence from our recent studies on opioid receptor dimerization. We believe a thorough understanding of receptor dimerization is crucial in clarifying the mechanism of action of opioids and other drugs and may serve a more practical purpose in aiding the development of novel therapeutic drugs. PMID- 11008064 TI - Molecular aspects of muscarinic receptor dimerization. AB - The five muscarinic acetylcholine receptors (M(1)-M(5)) are prototypical members of the superfamily of G-protein-coupled receptors (GPCRs). During the past decade, this laboratory has used different members of this receptor subfamily as model systems to study the molecular mechanisms involved in GPCR function. This article reviews recent investigations dealing with molecular aspects of M(3) muscarinic receptor assembly and dimerization/oligomerization. Studies with coexpressed M(3) receptor fragments and M(3) muscarinic/alpha(2C)-adrenergic hybrid receptors showed that muscarinic receptors, like other GPCRs, are composed of multiple autonomous folding domains. Moreover, biochemical studies have provided direct evidence for the formation of M(3) receptor dimers/oligomers. These high molecular mass receptor species are expressed on the cell surface and can bind muscarinic ligands. M(3) receptor dimerization/oligomerization appears to be receptor subtype-selective and involves both non-covalent interactions as well as disulfide-crosslinking of receptor monomers. These studies add to the growing number of reports suggesting the existence of GPCR dimers or multimers. The precise functional characteristics of such receptor aggregates remain to be elucidated. PMID- 11008065 TI - Oligomerization of dopamine and serotonin receptors. AB - Until recently, it has largely been assumed that G protein-coupled receptors (GPCRs) function as monomeric entities. However, over the past few years, we and others have documented that GPCRs can form dimers and oligomers, leading to a re evaluation of the mechanisms thought to mediate GPCR function. Despite the growing number of investigations into dimerization, little is known about the structural basis of receptor-receptor interactions and the functional consequences of dimer formation. Here, we present a brief review of some insights we have gained into the dimerization of dopamine and serotonin receptors. We have demonstrated that agonist-regulated trafficking is identical for receptor monomers and dimers, however, agonist treatment appears to stabilise the receptor oligomers. An investigation of the structural assembly between receptors involved in dimerization showed that there are several sites of interaction including hydrophobic transmembrane domain interactions and intermolecular disulphide bonds. We have also examined receptor hetero-oligomerization and demonstrated the potential for novel functions as a result of these associations. Finally, as a result of these observations, we have been able to present evidence that GPCRs function as oligomers in the cell. PMID- 11008066 TI - Signal transduction by GABA(B) receptor heterodimers. AB - GABA(B) receptors are G-protein-coupled receptors that mediate inhibition throughout the central and peripheral nervous systems. A single cloned receptor, GABA(B)R1, which has at least three alternatively spliced forms, appears to account for the vast majority of binding sites in the brain for high-affinity antagonists. In heterologous expression systems GABA(B)R1 is poorly expressed on the plasma membrane and largely fails to couple to ion channels. A second gene, GABA(B)R2, which exhibits moderately low homology to GABA(B)R1, permits surface expression of GABA(B)R1 and the appearance of baclofen-sensitive K(+) and Ca(+1) currents. We review the data that supports a model of the native GABA(B) receptor as a heterodimer composed of GABA(B)R1 and GABA(B)R2 proteins. New data from mutagenesis experiments are presented that point to amino acid residues on GABA(B)R1 critical for ligand activation of the heterodimer. The possible role of GABA(B)R2 in signal transduction is also discussed. The interdependent nature of the two subunits for receptor function makes the GABA(B) receptor a useful model to explore the larger significance of GPCR dimerization for G-protein activation. PMID- 11008067 TI - Evidence for adenosine/dopamine receptor interactions: indications for heteromerization. AB - Evidence has been obtained for adenosine/dopamine interactions in the central nervous system. There exists an anatomical basis for the existence of functional interactions between adenosine A(1)R and dopamine D(1)R and between adenosine A(2A) and dopamine D(2) receptors in the same neurons. Selective A(1)R agonists affect negatively the high affinity binding of D(1) receptors. Activation of A(2A) receptors leads to a decrease in receptor affinity for dopamine agonists acting on D(2) receptors, specially of the high-affinity state. These interactions have been reproduced in cell lines and found to be of functional significance. Adenosine/dopamine interactions at the behavioral level probably reflect those found at the level of dopamine receptor binding and transduction. All these findings suggest receptor subtype-specific interactions between adenosine and dopamine receptors that may be achieved by molecular interactions (e.g., receptor heterodimerization). At the molecular level adenosine receptors can serve as a model for homomeric and heteromeric protein-protein interactions. A1R forms homodimers in membranes and also form high-order molecular structures containing also heterotrimeric G-proteins and adenosine deaminase. The occurrence of clustering also clearly suggests that G-protein- coupled receptors form high order molecular structures, in which multimers of the receptors and probably other interacting proteins form functional complexes. In view of the occurrence of homodimers of adenosine and of dopamine receptors it is speculated that heterodimers between these receptors belonging to two different families of G protein-coupled receptors can be formed. Evidence that A1/D1 can form heterodimers in cotransfected cells and in primary cultures of neurons has in fact been obtained. In the central nervous system direct and indirect receptor receptor interactions via adaptor proteins participate in neurotransmission and neuromodulation and, for example, in the establishment of high neural functions such as learning and memory. PMID- 11008068 TI - Dimerization and domain swapping in G-protein-coupled receptors: a computational study. AB - In recent years there has been an increasing number of reports describing G protein-coupled receptor (GPCR) dimerization and heterodimerization. However, the evidence on the nature of the dimers and their role in GPCR activation is inconclusive. Consequently, we present here a review of our computational studies on G protein-coupled receptor dimerization and domain swapping. The studies described include molecular dynamics simulations on receptor monomers and dimers in the absence of ligand, in the presence of an agonist, and in the presence of an antagonist (or more precisely an inverse agonist). Two distinct sequence-based approaches to studying protein interfaces are also described, namely correlated mutation analysis and evolutionary trace analysis. All three approaches concur in supporting the proposal that the dimerization interface includes transmembrane helices 5 and 6. These studies cannot distinguish between domain swapped dimers and contact dimers as the models used were restricted to the helical part of the receptor. However, it is proposed that for the purpose of signalling, the domain swapped dimer and the corresponding contact dimer are equivalent. The evolutionary trace analysis suggests that every GPCR family and subfamily (for which sufficient sequence data is available) has the potential to dimerize through this common functional site on helices 5 and 6. The evolutionary trace results on the G protein are briefly described and these are consistent with GPCR dimerization. In addition to the functional site on helices 5 and 6, the evolutionary trace analysis identified a second functional site on helices 2 and 3. Possible roles for this site are suggested, including oligomerization. PMID- 11008069 TI - Expression of proinflammatory cytokines in the failing human heart: comparison of recent-onset and end-stage congestive heart failure. AB - BACKGROUND: Plasma levels of proinflammatory cytokines, including tumor necrosis factor (TNF)-alpha and interleukin (IL)-6, are elevated in patients with congestive heart failure (CHF). Recent studies suggest that the failing human heart is a source of proinflammatory cytokines in the end-stage failing heart. However, the relevance of plasma levels to those of the myocardium remains undefined. We sought to compare cytokine expression in early and end-stage CHF, and to evaluate the correlation of tissue expression to plasma levels. METHODS: Two patient populations were studied: patients with recent-onset CHF, all with symptoms less than 6 months (n = 17, duration of symptoms 2.1 +/- 1.6 months, range of New York Heart Association (NYHA) 1 to 3), and end-stage heart-failure patients (n = 7) who underwent left-ventricular assist-device (LVAD) implantation (Duration of symptoms 47.1 +/- 28.0 months, all NYHA class 4). Plasma levels of TNF-alpha and IL-6 proteins were evaluated by an Enzyme-Linked Immuno-Sorbent Assay (ELISA), while myocardial levels of cytokine transcripts were assessed by ribonuclease (Rnase) protection assay. RESULTS: In patients with end-stage heart failure, TNF-alpha and IL-6 were increased in the plasma as well as in the myocardium (plasma: TNF-alpha = 7.7 +/- 2.3 pg/ml, IL-6 = 45.0 +/- 47.1 pg/ml; myocardium: TNF-alpha = 0.31 +/- 0.15% of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression, IL-6 = 1.56 +/- 1.54% ). In contrast, despite elevated plasma levels of TNF-alpha and IL-6, the myocardium of patients with the recent onset of symptoms demonstrated minimal expression of TNF-alpha and IL-6 messenger ribonucleic acid (mRNA) (plasma: TNF-alpha = 4.3 +/- 1.7 pg/ml, IL-6 = 3.3 +/- 1.8 pg/ml; myocardium: TNF-alpha = 0.13 +/- 0. 04%, IL-6 = 0.02 +/- 0.04%). Plasma levels of TNF-alpha were significantly correlated with those in the myocardium when both populations were combined. (r = 0.69, p < 0.001). CONCLUSIONS: Cytokines are expressed in the myocardium in end-stage heart failure to a much greater degree than in patients with the recent-onset of symptoms. This suggests that induction of cytokines in the myocardium is a relatively late event in the pathogenesis of CHF. Furthermore, plasma levels of TNF-alpha correlates with mRNA expression in the myocardium and thus may serve as an appropriate marker of myocardial cytokine activation. Whether the production of cytokines in the failing human heart precedes the elevation of cytokines in the plasma remains undefined. Therefore, we studied expression of TNF-alpha and IL-6 in the myocardium as well as in the plasma in patients with early and end-stage CHF. The results have demonstrated that cytokines are expressed in the myocardium in end stage heart failure to a much greater degree than in patients with the recent onset of symptoms. This suggests that induction of cytokines in the myocardium is a relatively late event in the pathogenesis of CHF. PMID- 11008070 TI - Growth and pubertal development following pediatric heart transplantation: a 15 year experience at Ste-Justine Hospital. AB - BACKGROUND: Thirty-one children and adolescents have undergone allograft heart transplantation at Ste-Justine Hospital from July 1984 to August 1996. Twenty five patients were followed prospectively more than 3 years to document their growth and pubertal development. METHODS: Parameters surveyed were clinical (height, weight, pubertal staging, and bone age) and biochemical (luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, estradiol, dehydroepiandrosterone sulphate (DHEAS), IGF-1, and fasting insulin). RESULTS: At surgery, there were 18 boys and 7 girls aged 11 months to 17 years (median 13 years); 14 had congenital heart defects (CHDs) and 11 had a cardiomyopathy (CM). Immunosuppressive therapy included cyclosporine, azathioprine, and prednisone. Eighteen patients were still growing (15 boys, 3 girls): 8 had a retarded bone age and 6 with CHD had severe growth failure. Following surgery, most patients maintained their height within one sodium dodecyl sulfate (SDS) score of that initially observed. Patients reaching their target heights do so mainly in the lower range. Three patients not reaching target height had a CHD. Weight was greatest 1 year postoperatively (113 +/- 27% ideal body weight) with normalization at 2 years (100 +/- 18%). Of the 13 prepubertal patients, menarche occurred at age 12 in 1 girl, while 3 boys began puberty at age 12 years. In both sexes, serum levels of gonadotropins and IGF-1 increased during puberty, moderate hyperinsulinism was observed, and DHEAS levels decreased. CONCLUSIONS: Our results indicate that children and adolescents grow normally following cardiac transplantation and that they attain their target height despite a lack of catch up growth. They gain weight significantly in the first postoperative year with normalization of their weight at 2 years. Furthermore, the clinical and biochemical indices of puberty are overall within the norms. However, the severity of growth delay at the time of transplantation inherent to the cardiac pathology has a major impact on adult height. PMID- 11008071 TI - Utility of extracorporeal membrane oxygenation for early graft failure following heart transplantation in infancy. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is widely used for postcardiotomy cardiogenic shock in children. However, the efficacy of ECMO for early post-heart transplant graft failure in infants has not been reported. Our aims were to determine: (1) the utility of ECMO in infants with severe donor heart dysfunction, (2) predictors for requiring ECMO, and (3) the long-term outcome of surviving ECMO patients. METHODS: All infants (age < 6 months at listing) undergoing heart transplantation were reviewed. Diagnostic categories were hypoplastic left heart syndrome (HLHS) and non-HLHS (complex congenital heart disease and cardiomyopathies). Continuous and categorical comparisons were by Wilcoxon's rank sum test and Fisher's exact test respectively. RESULTS: 14 (12 HLHS, 2 non-HLHS) of 63 (46 HLHS, 17 non-HLHS) infants were placed on ECMO. Ten patients (71%) were successfully weaned from ECMO and 8 (57%) were discharged alive. All ECMO hospital survivors remain alive (mean follow-up 36.2 +/- 21.4 months, range 13.1-77.6 months). Mean duration of ECMO support was 68 hours in weaned patients vs 144 hours (p = 0.19) in nonweaned patients, and 64 hours in survivors vs 123 hours (p = 0.35) in nonsurvivors. ECMO deaths were due to sepsis (n = 3), intractable pulmonary hypertension (n = 2), and intracranial bleed (n = 1). Neurologic deficits occurred in 2 survivors. Median ICU and hospital stays for ECMO survivors were 29 and 33 days vs 7 (p = 0.0003) and 9 (p = 0.0004) days for non-ECMO patients. Age listed, age transplanted, wait time, body weight, donor/recipient weight ratio, total ischemia time, and diagnosis did not predict the need for ECMO. CONCLUSIONS: (1) ECMO is useful for post-heart transplant circulatory support in infants with early graft failure. (2) All survivors were weaned in fewer than 4 days. (3) Three-year survival of ECMO hospital survivors has been high, but neurologic complications are prevalent. PMID- 11008072 TI - Glucocorticoid-induced osteopenia in adolescent heart transplant recipients. AB - BACKGROUND: Glucocorticoid-induced cushingoid symptoms, including osteopenia and osteoporosis are well-documented in adult heart transplant recipients (HTR). Bone mineral density (BMD) of the axial skeleton is diminished by 10% to 20% within 60 days after transplantation (Tx) and most adult HTR fulfill World Health Organization criteria for osteoporosis (BMD > 2.5 SD below norm). At present, we do not know whether glucocorticoids have similar deleterious effects in adolescent HTR. METHODS: To determine the consequences of glucocorticoid immunosuppression on regional bone mineral density (BMD) and biochemical markers of bone metabolism in adolescent HTR, we studied 19 patients (aged 16 +/- 3) at 19 months (group mean) after Tx. We measured BMD (hydroxyapatite g/cm(2)) of the total body, lumbar spine, and pelvis using dual-energy X-ray absorptiometry (Lunar). Serum levels of bone-specific alkaline phosphatase and pyridinoline cross-links were determined by enzyme immunoassay in serum kits. RESULTS: The BMD of the lumbar spine (-12%), femur neck (-13%), femur trochanter (-12%), and ward's triangle (-16%) were significantly (p < 0.05) lower in adolescent HTR than age- and gender-matched norms. Serum levels of alkaline phosphatase (29 +/- 6 vs 22 +/- 3 U/liter) and pyridinoline cross-links (5.3 +/- 1.1 vs 3.8 +/- 0.7 mmol/liter) were significantly (p < 0.05) elevated in adolescent HTR, compared with age- and gender-matched controls studied in our laboratory. CONCLUSIONS: Our cross-sectional results demonstrate that BMD of the axial skeleton in adolescent HTR is significantly lower (-10% to 20%) than age-matched norms and that serum biochemical markers of bone metabolism are significantly elevated, suggesting accelerated bone turnover. PMID- 11008073 TI - Lung allocation in the United States, 1995-1997: an analysis of equity and utility. AB - BACKGROUND: Waiting time for organ transplantation varies widely between programs of different sizes and by geographic regions. The purpose of this study was to determine if the current lung-allocation policy is equitable for candidates waiting at various-sized centers, and to model how national allocation based solely on waiting time might affect patients and programs. METHODS: UNOS provided data on candidate registrations; transplants and outcomes; waiting times; and deaths while waiting for all U.S. lung-transplant programs during 1995-1997. Transplant centers were categorized based on average yearly volume: small (< or = 10 pounds sterling transplants/year; n = 46), medium (11-30 transplants/year; n = 29), or large (>30 transplants/year; n = 6). This data was used to model national organ allocation based solely on accumulated waiting time for candidates listed at the end of 1997. RESULTS: Median waiting time for patients transplanted was longest at large programs (724-848 days) compared to small and medium centers (371-552 days and 337-553 days, respectively) and increased at programs of all sizes during the study period. Wait-time-adjusted risk of death correlated inversely with program size (365 vs 261 vs 148 deaths per 1,000 patient-years-at risk at small, medium, and large centers, respectively). Mortality as a percentage of new candidate registrations was similar for all program categories, ranging from 21 to 25%. Survival rates following transplantation were equivalent at medium-sized centers vs large centers (p = 0.50), but statistically lower when small centers were compared to either large- or medium-size centers (p < or = 0.05). Using waiting time as the primary criterion lung allocation would acutely shift 10 to 20% of lung-transplant activity from medium to large programs. CONCLUSIONS: 1) Waiting list mortality rates are not higher at large lung transplant programs with long average waiting times. 2) A lung-allocation algorithm based primarily on waiting-list seniority would probably disadvantage candidates at medium-size centers without improving overall lung-transplant outcomes. 3) If fairness is measured by equal distribution of opportunity and risk, we conclude that the current allocation system is relatively equitable for patients currently entering the lung-transplant system. PMID- 11008074 TI - The utility of open lung biopsy following lung transplantation. AB - BACKGROUND: Most pulmonary complications associated with lung transplantation have non-specific clinical characteristics. Furthermore, common diagnostic modalities, including bronchoscopy with transbronchial biopsy (TBB), often do not render a definitive diagnosis. In this study, we reviewed our experience with open lung biopsy (OLB) following lung transplantation, specifically regarding its ability to safely provide clinically relevant information that affects therapeutic decisions. METHODS: From October 1989 to March 2000, 202 patients underwent lung transplantation at our institution. We reviewed the clinical course of the 42 patients who received 48 OLBs. Of these patients, we determined the pre-operative clinical condition, preceding TBB histologic information, OLB histology, treatment changes, and procedural complications as a result of the OLB. RESULTS: A new, clinically unsuspected diagnosis was made in 14 biopsies (29% of all OLB), and all of these resulted in therapy changes. Thirty-two biopsies (67% of all OLB) confirmed our clinical suspicions, and new therapy was initiated in 30 of these patients. Two patients (4% of all OLB) had non diagnostic OLB. Four biopsies (8% of all OLB), including the 2 non-diagnostic OLBs, did not result in any therapy changes or initiation of new therapy. Complications occurred in 3 patients, all of whom had an air leak for >7 days. CONCLUSION: Open lung biopsy in lung transplant patients renders a new, unsuspected diagnosis in nearly one third of patients and leads to specific, directed therapy in the vast majority of patients. Open-lung biopsy can be performed safely and should be considered when diagnosis is uncertain in clinically deteriorating patients. PMID- 11008075 TI - Increased expression of inflammatory cytokines and adhesion molecules by alveolar macrophages of human lung allograft recipients with acute rejection: decline with resolution of rejection. AB - BACKGROUND: Alveolar macrophages (AM) are the major population in bronchoalveolar lavage (BAL) cells; we assessed their role in human lung allograft recipients by correlating the expression of adhesion molecules and inflammatory cytokines with clinical outcome of allograft. METHODS: We obtained BAL samples from patients and enriched them for AM in plastic petri dish for 2 hours at 37 degrees C in 5% CO(2). Expression of intercellular adhesion molecule-1 (ICAM-1, CD54), platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31), and CD11c was assessed by flow cytometry using monoclonal antibodies. We assessed cytokine profile using Multi-Probe RNase protection assay. RESULTS: Alveolar macrophages that express CD11c, CD31 and CD54 were increased in patients with either rejection or infection compared with those without rejection and infection. The difference in the percentage of AM expressing CD11c and CD31 between the rejection group and patients without rejection and infection group was statistically significant (CD11c, p < 0.01; CD31, p < 0.03). Interleukin (IL)-1 alpha, IL-1 beta, IL-1 receptor antagonist (IL-1Ra), and IL-6 expression was higher in the rejection group than in patients without rejection. Five out of 9 patients in the rejection group expressed high levels of IL-15 and tumor necrosis factor-alpha compared with patients without rejection and infection. The increased number of AM expressing adhesion molecules and elevated expression of cytokines observed during acute rejection declined to basal levels after successful treatment and resolution of rejection. This study demonstrates that lung allograft rejection is associated with increased expression of adhesion molecules and inflammatory cytokines by AM, which could facilitate mononuclear cell adhesion and extravasation contributing to the allograft injury in lung transplant recipients. PMID- 11008076 TI - Renal failure after clinical heart transplantation is associated with the TGF beta 1 codon 10 gene polymorphism. AB - BACKGROUND: To determine whether genetic factors are involved in the development of renal dysfunction due to cyclosporine nephrotoxicity, we analyzed 2 polymorphisms in the signal sequence of the transforming growth factor (TGF)-beta 1 gene; codon 10 (Leu(10) --> Pro) and codon 25 (Arg(25) --> Pro). METHOD: Using sequence specific oligonucleotide probing, we analyzed both TGF-beta1 gene polymorphisms in cardiac allograft recipients (n = 168) who survived at least 1 year with minimal follow-up of 7 years. Patients received cyclosporine and steroids as maintenance immunosuppressive therapy. Renal dysfunction was defined as a serum creatinine > or = 250 micromol/liter. RESULTS: Renal dysfunction was observed in 2% (3/168) of the patients at 1 year, in 7% (11/160) at 3 years, in 12% (18/152) at 5 years, and in 20% (26/131) at 7 years post-transplantation. The genotypic distributions for TGF-beta1 codon 10 were 7% Pro/Pro, 61% Pro/Leu, and 32% Leu/Leu, and for codon 25 these percentages were 1% Pro/Pro, 12% Pro/Arg, and 87% Arg/Arg. We found an association between the TGF-beta 1 genotype encoding proline at codon 10 and renal dysfunction. At 7 years post-transplantation, 26% (23/89) of the patients with the heterozygous Pro/Leu or homozygous Pro/Pro genotype had renal dysfunction vs only 7% (3/42) of the patients with the homozygous Leu/Leu genotype (p = 0.017). For the TGF-beta1 codon 25 genotypes, we found no association between TGF-beta 1 genotypes and renal dysfunction. CONCLUSION: Our data support the hypothesis that TGF-beta 1 is involved in the process leading to renal insufficiency in cyclosporine-treated cardiac allograft recipients. In these patients the presence of TGF-beta 1 Pro(10) might be a risk factor. PMID- 11008077 TI - Myocardial perfusion scintigraphy as a screening method for significant coronary artery stenosis in cardiac transplant recipients. AB - BACKGROUND: Several studies have explored the feasibility of using myocardial perfusion imaging to detect allograft vasculopathy after heart transplantation. We undertook the present prospective consecutive study to comparatively evaluate the role of serial myocardial perfusion single-photon emission computed tomography (SPECT) scanning and coronary arteriography (CAG) in detecting coronary artery stenosis suitable for coronary angioplasty in heart transplant recipients. METHODS: Within a 2-week interval during a follow-up period of 5.6 (95% confidence limits 2.1 to 12) years, 255 serial CAGs and myocardial perfusion scintigraphies were performed in 67 patients. Arteriography and scintigraphy were performed once yearly after heart transplantation. We retrospectively analyzed the data. RESULTS: Myocardial scintigraphy showed pathologic reversible defects in 9 out of 67 patients. Four of these patients had significant (>50% and also >70%) focal segmental stenosis in the middle and proximal parts of the coronary arteries (Type A lesions), 1 had diffuse and circumferential narrowing in the distal parts (Type B lesions), whereas CAG showed no lesions in the remaining 4 patients. The patients with significant Type A lesions were revascularized with percutaneous coronary angioplasty. Coronary arteriography showed that 1 patient had extensive Type A and Type B lesions, whereas myocardial perfusion scans detected no. The predictive value of a negative (normal) SPECT was 98% (95% confidence limits 94% to 100%) for the detection of lesions suited for revascularization. CONCLUSIONS: Annual myocardial SPECT seems well suited to screen for significant coronary artery stenosis. A SPECT study without reversible defects virtually excludes lesions suitable for coronary artery revascularization. PMID- 11008078 TI - FR167653 ameliorates pulmonary damage in ischemia-reperfusion injury in a canine lung transplantation model. AB - BACKGROUND: Interleukin (IL)-1 and tumor necrosis factor-alpha (TNF-alpha) are recognized as important factors in ischemia-reperfusion (I/R) injury. FR167653 has been characterized as a potent suppressant of IL-1 and TNF-alpha production. We previously reported that FR167653 suppressed the expression of IL-1 beta mRNA after reperfusion and ameliorated pulmonary I/R injury following 3-hour left lung warm ischemia in dogs. The aim of this study was to investigate the effects of FR167653 on I/R injury in a canine left, single, lung transplantation model. METHODS: We used 10 pairs of weight-matched dogs. We assigned 5 pairs to the FR group, in which each animal received FR167653 (1 mg/kg/hr) IV from 30 minutes before ischemia until 2 hours after reperfusion; we treated the transplanted lungs with FR167653 after the onset of reperfusion. The others were assigned to the control group. After 8-hour preservation with 4 degrees C Euro-Collins solution, orthotopic left, single, lung transplantation was performed. During a 5 minute clamping test at the right pulmonary artery of each recipient, the left (transplanted) pulmonary arterial pressure (L-PAP), left (transplanted) pulmonary vascular resistance (L-PVR), arterial oxygen pressure (PaO(2)), and alveolar arterial oxygen pressure difference (A-aDO(2)) were measured. We harvested transplanted lung specimens for histologic study, and we counted polymorphonuclear neutrophils (PMNs), which were identified by staining with naphthol AS-D cholroacetate esterase. Pulmonary perfusion and ventilation scintigraphy (Tc-99m-MAA and Xe-133) were performed. We observed the animals for 3 days after transplantation. RESULTS: The PAP, L-PVR, PaO(2), and A-aDO(2) revealed significantly (p < 0.05) better function in the FR group than in the control group. Histologically, lung edema was milder, and PMN infiltration was significantly (p < 0.05) lower in the FR group than in the control group. Xe-133 and Tc-99m-MAA were widely distributed throughout the graft lung in the FR group. Three-day survival rates in FR and control groups were 60% and 20%, respectively. CONCLUSIONS: FR167653 appears to generate a protective effect on I/R injury in lung transplantation in dogs. PMID- 11008079 TI - Effects of EP4 solution and LPD solution vs Euro-Collins solution on Na(+)/K(+) ATPase activity in rat alveolar type II cells and human alveolar epithelial cell line A549 cells. AB - BACKGROUND: Intact alveolar epithelial Na(+)/K(+)- adenosinetriphosphatase (ATPase) function is important in preventing alveolar fluid accumulation after lung transplantation. We examined whether the type of preservation solution used influences Na(+)/K(+)-ATPase activity in alveolar epithelial cells. METHODS: Rat alveolar type II cells were preserved with EP4, low-potassium dextran (LPD), or Euro-Collins solution at 7 degrees C for 5 and 20 hours. To assess cell toxicity, we measured cell viability and lactate dehydrogenase release. Na(+)/K(+)-ATPase activity was measured as ouabain-sensitive ATPase hydrolysis. We also examined the effect of terbutaline (10(-3) mol/liter) and dibutyryl cyclic adenosine monophosphate (dbcAMP) (10(-3) mol/liter) on Na(+)/K(+)-ATPase activity in A549 cells preserved for 5 hours. RESULTS: All solutions caused significant damage of rat alveolar type II cells at 20 hours. However, Na(+)/K(+)-ATPase activity was preserved at normal levels with EP4 and LPD over 20 hours. Terbutaline and dbcAMP significantly increased Na(+)/K(+)-ATPase activity in A549 cells preserved with EP4 and LPD solutions for 5 hours. However, we observed no activation in the cells preserved with Euro-Collins solution. We found no significant difference in intracellular cAMP levels after terbutaline challenge among the types of preservation solution. CONCLUSIONS: We conclude that extracellular-type solutions such as EP4 and LPD may be preferable for maintaining not only the basal activity but also the ability to activate Na(+)/K(+)-ATPase in response to beta-adrenergic agonists, in alveolar epithelial cells. PMID- 11008080 TI - Lung transplantation from dialysis dependent donors. AB - Lung transplantation from a donor with chronic renal failure has never been reported. This paper reports our successful experience with 2 transplants from donors with end-stage renal disease who were on chronic hemodialysis, and reviews the relevant literature on the effects of renal failure on pulmonary function and on the use of marginal donors. PMID- 11008081 TI - Orthotopic heart transplantation with concurrent aortic valve replacement and coronary artery bypass grafting. AB - We report a case of successful orthotopic heart transplantation of a donor heart with normal ventricular function, 2-vessel coronary artery disease, and a bicuspid aortic valve, which required concurrent aortic valve replacement and coronary artery bypass grafting. In confronting the disparities in demand and supply, we must consider the so-called marginally acceptable heart for either critically ill recipients or those who may be disadvantaged on the waiting list. PMID- 11008082 TI - Dactylaria gallopava infection presenting as a pulmonary nodule in a single-lung transplant recipient. AB - We describe the first case of a pulmonary nodule caused by Dactylaria gallopava in a lung-transplant recipient. An asymptomatic lung-allograft recipient was found to have a 2-cm nodule in the native lung 450 days after transplantation. Culture of a transthoracic needle biopsy of the solitary pulmonary nodule revealed Dactylaria gallopava. Treatment was initiated with amphotericin B for a period of 21 days followed by oral itraconazole for an 8-month period. PMID- 11008083 TI - Isoniazid prophylaxis in lung transplantation. AB - The incidence of tuberculous disease (TD) is higher in lung-transplant patients than in the general population. During a 7-year period, we included 61 patients who underwent lung transplantation in a prospective isoniazid prophylaxis protocol. Isoniazid was prescribed to infected and anergic patients not previously treated when added to the waiting list. Six of 61 patients (10%) developed tuberculosis. We observed no differences in tuberculous disease incidence between infected-anergic and non-infected patients. In our tuberculous endemic area, isoniazid prophylaxis is safe and offers protection from TD to infected and anergic patients who must be enrolled in a lung transplantation program. PMID- 11008084 TI - Research and practice in adolescent medicine: "both-and" not "either-or" for clinicians. PMID- 11008085 TI - Social cognitions associated with pubertal development in a sample of urban, low income, African-American and Latina girls and mothers. AB - PURPOSE: To assess girls' acquisition of new sex-related social cognitions at puberty and the social meanings they attribute to changes in their primary social relationships. METHODS: As part of a larger study assessing sociosexual cognitions of urban girls, 57 African-American and Latina mothers and 57 girls (aged 10-13 years) participated in one of 16 focus groups. Thematic analyses were conducted on transcripts of the mothers' and daughters' focus group sessions. RESULTS: Analyses revealed four major themes related to pubertal development: (1) physical maturation provides new social status of maturity; (2) puberty is associated with changes in sexual expectations and roles; (3) girls develop social meanings to feelings of sexual arousal; and (4) puberty prompts changes in mother-daughter relationship control. Representative quotations are used to illustrate each of these themes. CONCLUSIONS: Our analyses provide insight into the ways in which late childhood and early adolescent urban girls interpret sexuality and sexual relations, perceive changes in their social relationships, and develop expectations regarding their roles in sexual and romantic relationships. PMID- 11008086 TI - Postponing sexual intercourse among urban junior high school students-a randomized controlled evaluation. AB - PURPOSE: To describe a randomized, controlled evaluation of a school-based intervention to delay sexual intercourse among urban junior high school students. METHODS: Six Washington, D.C., junior high schools were randomly assigned to the intervention or nonintervention control condition for an educational program. During the first school year, seventh graders (n = 582) from the six schools obtained written parental consent to participate. Three health professionals (one per intervention school) implemented the program, which consisted of reproductive health classes, the Postponing Sexual Involvement Curriculum, health risk screening, and "booster" educational activities during the following (eighth grade) school year. Cross-sectional surveys were administered at baseline, the end of the seventh grade, and the beginning and end of the eighth grade. Intervention and control group differences in virginity, attitudes toward delayed sex and childbearing, and sexual knowledge and behavior were assessed at all four time points. RESULTS: At baseline, 44% of the seventh grade males and 81% of the seventh grade females reported being virgins. At the end of the seventh grade (first follow-up), after controlling for baseline study group differences, intervention-group females were more likely to report virginity, self-efficacy to refuse sex with a boyfriend, and the intention to avoid sexual involvement during the following 6 months. At the end of the eighth grade, significantly more intervention- than control-group females reported virginity, birth control use at last intercourse (for nonvirgins), and knowledge of adolescent reproductive health and confidentiality rights. No changes in virginity, self-efficacy to refuse sex, or sexual intent for the next 6 months were observed among male participants at any time during the study. However, on all three follow-up surveys, intervention-group males scored significantly higher than their control group counterparts in knowledge of birth control method efficacy. No change in attitudes toward abstinence was observed for either gender at any follow-up point. CONCLUSIONS: Gender differences in baseline sexual activity rates and in various study outcomes suggest a possible need for separate, gender-specific intervention activities that can adequately address the social and cognitive needs of both sexes. PMID- 11008087 TI - Exposure to, and perceived usefulness of, school-based tobacco prevention programs: associations with susceptibility to smoking among adolescents. AB - PURPOSE: To examine the relationships of exposure to school-based tobacco prevention programs and perceived usefulness of such programs to smoking susceptibility. METHODS: An ethnically representative sample of 6929 Grade 10 students participated in a cross-sectional survey of tobacco-related activities and behaviors. Perceived usefulness of prevention programs was investigated according to two dimensions: perceived helpfulness of information received at school with regard to making decisions about tobacco use and perceived availability of social resources that might help students cope with tobacco related issues. RESULTS: The extent of exposure to school prevention programs and both dimensions of perceived usefulness of these programs were inversely associated with susceptibility to smoking. In addition, perceived information helpfulness mediated the effect of program exposure on susceptibility status, whereas perceived availability of social resources moderated that effect. The study also presented evidence suggesting ethnic/cultural variability in the status of susceptibility to smoking at this age. CONCLUSIONS: The study indicates a need to examine the perceptions of students receiving school prevention programs. Perceived usefulness of school prevention programs may be used to identify youth at high risk for smoking susceptibility. Future studies, however, should investigate other dimensions of perceived usefulness as well as intrapersonal, interpersonal, and environmental correlates of this construct. PMID- 11008088 TI - Psychosocial determinants of the onset and escalation of smoking: cross-sectional and prospective findings in multiethnic middle school samples. AB - PURPOSE: To investigate a broad range of social influence-related and global determinants of smoking to aid in the design of comprehensive multiethnic interventions by testing the most important factors of initiation and escalation of smoking across various subgroups. METHODS: Cross-sectional (N = 2546) and cohort (N = 736) samples of multiethnic middle school students near a large Southwestern metropolis were surveyed through self-report questionnaires. The confidential questionnaires included information on demographics, risk factors, and smoking behavior and were administered in class by trained data collectors. Multivariable logistic regression analysis was used to examine the statistical significance and strength of the factors. RESULTS: Those lower in self-esteem and higher in social assertiveness appeared to be most at risk for the onset of smoking, whereas those low in optimism appeared to be the most at risk for the escalation of smoking. Attitudes, friends' norms, parents' norms, perceived behavioral control, and perceived prevalence were consistent predictors of all smoking status outcomes. CONCLUSIONS: The behavioral-specific determinants of smoking appear to be important predictors of smoking status outcomes in all demographic subgroups. The relationships of the global determinants were more dependent on the smoking outcome variable and subgroup examined. The findings may serve to help facilitate the targeting of comprehensive interventions aimed at reducing adolescent smoking in multiethnic and ethnic group-specific populations. PMID- 11008089 TI - Prevalence of adolescent injury from recreational exercise: an Australian perspective. AB - PURPOSE: To report the prevalence of recent adolescent recreational and sporting activities and associated injury. METHODS: Data were collected during three school terms in 1997 using a validated questionnaire administered once only to 3538 girls and boys aged 11-12 years and 15-16 years. These students comprised 97.5% of the students in these age ranges in randomly selected state and private schools in the Adelaide metropolitan area (South Australia). Participants identified up to three recreational and/or sporting activities in which they had participated in the previous week. Data were collected on the nature and extent of participation, and on associated injuries. Participation and injury reports were summarized descriptively in gender strata in the two adolescent age groups and stratum specific odds ratios were used to explore injury risk. RESULTS: Subjects reported participating in 8997 sporting and/or recreational activities in the preceding week (an average of 2.5 participations per student). Over 140 sports and recreational pursuits were represented, incorporating organized and nonorganized activities undertaken in teams, social groups, or alone. Approximately 25% of adolescents reported at least one recreational injury. Injuries were mostly minor, reflecting soft tissue trauma and skin abrasions. Organized group sport incurred the highest risk of injury. There were marked gender and year level differences in injury risk in a number of common activities. CONCLUSIONS: These findings support the need for ongoing education regarding injury prevention and management. PMID- 11008090 TI - Reliability of adolescents' self-report of recent recreational injury. AB - PURPOSE: To assess the reliability of adolescents' self-report of recent recreational injury. METHOD: Identical written questionnaires were administered twice in a 3-week period to 75 randomly selected adolescents (aged 11-12 years and 15-16 years) in state primary and secondary schools in South Australia, Australia. The questionnaires sought information on injury sustained in the previous week. Parents of 60 of the adolescents were contacted in the same week as one of the questionnaire administrations, to verify their adolescent's most recent injury self-report. Intraclass and Pearson correlation coefficients provided evidence of agreement between responses, and stability of measure. RESULTS: There was high stability and good agreement between the parent and adolescent responses provided in the same week, suggesting that adolescents accurately reported recent recreational injury. The poor stability and low agreement between repeated adolescent responses over the 3-week period indicated not poor recall, but the changing nature of mostly minor recreational injury. These findings suggest that the nature of the injury reported in the first questionnaire had changed by the time of the second questionnaire administration, and that this was accurately reported. CONCLUSION: Confidence can be placed in adolescents' self-reports of recreational injury in the preceding week. PMID- 11008091 TI - Sexual practices in Malaysia: determinants of sexual intercourse among unmarried youths. AB - This paper describes findings on selected determinants of sexual intercourse among 468 unmarried adolescents from a survey in Malaysia. Data on respondents' background, sexual experience, contraceptive use, and sexual attitudes are provided. Based on multiple logistic regressions, factors significantly predictive of sexual experience are gender, employment, and sexual attitudes. PMID- 11008093 TI - Cytokine therapy in renal cell cancer. AB - Despite extensive investigations with many different treatment modalities, metastatic renal cell carcinoma (RCC) remains a disease highly resistant to systemic therapy. The outlook for patients with metastatic RCC is poor, with a 5 year survival rate of less than 10%. Late relapses after nephrectomy, prolonged stable disease in the absence of systemic therapy, and rare spontaneous regression are clinical observations that suggest host immune mechanisms could be important in regulating tumor growth. Interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) have been extensively studied in advanced RCC with responses in the 10 to 20% range. Two randomized trials suggest that treatment with IFN-alpha compared with vinblastine or medroxyprogesterone results in a small improvement in survival. Prolonged responses with high-dose IL-2 is significant but is accompanied by formidable toxicity. Although the combination of IFN-alpha and IL 2 compared with monotherapy with IFN-alpha or IL-2 increases the response proportion, no improvement in survival could be demonstrated in a recent randomized trial. In addition, three randomized trials showed no survival benefit associated with IFN-alpha therapy given as adjuvant therapy following complete resection of locally advanced RCC. Small numbers of patients exhibit complete or partial responses to IFN-alpha and/or IL-2, but most patients do not respond and there are few long-term survivors. Clinical investigation of new agents and treatment programs to identify improved antitumor activity against metastases remain the highest priorities in this refractory disease. PMID- 11008092 TI - Cigarette smoking in multicultural, urban high school students. AB - PURPOSE: To profile patterns of cigarette use among a multiethnic population of high school students, and identify important factors associated with cigarette use by ethnicity, in order to plan effective health promotion strategies. METHODS: This cross-sectional study involved the completion of a lifestyle questionnaire by 1236 Grade 9-13 students (86% response rate) from 62 randomly selected classrooms in three urban high schools in Toronto. Chi-square analysis of the association between tobacco use and other variables took account of the clustered sample using CSAMPLE in Epi Info. RESULTS: The students self-identified their ethnicity as follows: 388 Canadian, 269 European, 171 East Indian, 137 Asian, 76 West Indian, and 194 "other." Students who identified themselves as Canadians were significantly more likely to be current smokers (29%) than students reporting other ethnicities (13%). There was no apparent increase in smoking rates for immigrants after 2 or more years in Canada. Current drinking, sexual activity, and especially friends smoking was most strongly associated with current smoking for most ethnic groups, although the relative importance of these variables was not identical for all groups. CONCLUSIONS: Prevention programs may benefit from a focus on the influence of peer smoking and on the grouping together of lifestyle factors associated with smoking for students in all ethnic groups in this multicultural city. PMID- 11008094 TI - Prostate tumor progression and prognosis. interplay of tumor and host factors. AB - The prognosis for prostate cancer is largely dependent on the probability of metastatic dissemination. Prognostic markers currently in use are very poor predictors of metastatic potential, and as of yet none of the battery of new molecular markers has proven greatly superior. This may be due in part to their inability to assess the degree of interaction of subpopulations of prostate cancer cells with each other and with their microenvironment. A growing body of evidence indicates that these types of interactions are a major factor in the eventual genesis of cancer cells capable of metastasis. Recent research has demonstrated that specialized components of prostate tumors may play a critical supporting role for the overall growth of the larger tumor. The multifocal nature and apparent polyclonal origins of prostate tumors suggest that carcinogenesis and tumor progression are promoted by global influences or "field effects." It appears that these effects extend beyond the proliferating epithelial component to the tissue stroma. Prostate cancer cells and stromal cells seem to act in concert to modify the microenvironment, leading to metastasis. An understanding of this synergy may provide a new class of prognostic markers which more accurately measure the complex set of interactions that determine tumor behavior. PMID- 11008095 TI - Clinical trials of immunotherapy for advanced prostate cancer. AB - There is a lack of effective therapeutic regimens for advanced hormone-refractory prostate cancer (HRPC). Recent combination regimens of chemotherapy have improved management of HRPC. Neither systemic chemotherapy nor radiation regimens have significantly improved survival. Conventional systemic cytokine therapy has had limited efficacy in the treatment of advanced prostate cancer patients and its toxicity is severe. Combinations of multiple biological response modifiers for treatment of this disease also have limited efficacy. Results from phase II trials have shown that the combination of interferon-alpha and interleukin-2 therapy and the infusion of dendritic cells primed with peptides of prostate specific membrane antigen are promising. The former showed 31% response using the National Prostatic Cancer Project criteria, and the latter showed 27% of objective partial response with a reduction of >50% prostate specific antigen level. The toxicity of these two regimens was tolerated by patients. New approaches with tumor vaccines in conjunction with cytokine gene therapy have also been investigated. The clinical responses of these trials have been limited but promising. Immunotherapy may become an effective modality of prostate cancer treatment in the future. PMID- 11008096 TI - Comparative studies of prostate cancer in Japan versus the United States. A review. AB - This article reviews the available data on prostate cancer in Japan compared with that in the United States, with emphasis on epidemiologic, pathologic, and molecular aspects. Previous studies have demonstrated ethnic/racial differences in the incidence of prostate cancer between the two countries. Recent investigations indicate that different genetic alterations or polymorphisms are related to carcinogenesis in the prostate. Comparative geographic-pathologic autopsy studies suggest that different promoting factors including genetic, epigenetic, and environmental influences may be responsible for ethnic variations in the postinduction progression of prostate cancer. PMID- 11008097 TI - Quantitative Imaging of Yttrium-86 with PET. The Occurrence and Correction of Anomalous Apparent Activity in High Density Regions. AB - Purpose: Yttrium-86 has been proposed for use as a quantitative positron emission tomography imaging agent to determine the in vivo distribution of therapeutic pharmaceuticals labeled with yttrium-90, a pure beta minus emitter. This study identifies, and proposes a solution for, an artifact, which interferes with quantitation.Procedures: Yttrium-86 is a 14.7-hour halflife positron emitter (33% abundance) with multiple high energy gamma rays in cascade. Phantom measurements with a GE Advance PET scanner using standard attenuation and scatter corrections, demonstrated anomalous apparent activity in inactive higher density regions.Results: Apparent activity up to 30% of the surrounding true activity was observed in a bone equivalent material. Even higher activities were observed if the scatter correction was omitted. This phenomenon was determined to result from the effect of attenuation correction on true coincidences between one gamma ray and a second gamma ray or annihilation photon.Conclusion: A simple additional correction based on sinogram tail subtraction improves accuracy significantly. PMID- 11008098 TI - Evaluation of Abdominal Tumors with AB - Purpose: The purpose of this study was to evaluate the diagnostic usefulness of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET).Procedures: We performed FDG-PET in 22 patients with suspected primary or recurrent cancer (23 cases). The FDG-PET findings were compared with the final diagnosis, which was obtained by histological examination of resected, or biopsy specimens and the clinical course.Results: In all patients, the sensitivity, specificity, and the accuracy of FDG-PET were 93.8%, 85.7%, 91.3%, respectively. In the patients with colo-rectal cancer, the sensitivity, specificity, and accuracy were 92.9%, 100%, 94.4%, respectively.Conclusions: FDG-PET was considered to be clinically useful for the detection of primary or recurrent malignant tumors, and for differentiating recurrent tumors from inflammatory scars. PMID- 11008099 TI - Comparison of FDG-PET and Bone Scans for Detecting Skeletal Metastases in Patients with Non-small Cell Lung Cancer. AB - Purpose: Positron Emission Tomography (PET) with F18-fluorodeoxyglucose has been proven useful for staging non-small cell lung cancer. Bone scans are frequently performed for suspected skeletal metastases. The purpose of this study was to evaluate if bone scans compared to PET scans provide additional information that changes the stage of disease.Procedures: Nineteen patients with non-small cell lung cancer had PET and bone scans done for staging of the malignancy. The results of both studies were compared.Results: Bone and PET scans agreed on the presence or absence of skeletal metastases in all nineteen patients. The addition of a bone scan to a PET scan did not change the stage of the disease or the management in any of the patients. Bone scans allowed for more precise localization of the lesions in some patients.Conclusions: Bone scans do not change the stage of disease when performed in addition to PET scans, but provide more precise localization of skeletal abnormalities. PMID- 11008100 TI - Whole-Body FDG-PET in Patients with Recurrent Colorectal Carcinoma. A Comparative Study with CT. AB - Purpose: To assess the clinical accuracy of whole-body 2-[F-18]-fluoro-2-deoxy-D glucose-positron emission tomography (FDG-PET) in the diagnosis of recurrent colorectal carcinoma in comparison to conventional computed tomography (CT).Materials and methods: Forty patients with suspected recurrent colorectal carcinoma based on either progressive serial carcinoemrbyonic antigen (CEA) serum elevation or positive/equivocal CT findings underwent whole-body FDG-PET. PET results were compared with those of CT and correlated to the final histopathological and clinical findings.Results: A final diagnosis was obtained at 93 sites in 35 patients by histology and in 5 patients by clinical follow up of at least 6 months. Of the 93 sites, 53 were determined to be malignant and 40 benign. FDG-PET evaluated on a 5-point scale (0-4) showed a positive and negative predictive value in the range of 96-98% and 83-93% respectively as the threshold for positivity was moved from 0 through 3. By comparison, CT, also evaluated on a 5-point scale showed a positive and negative predictive value in the range of 75 88% and 67-71% respectively. The area under the fitted receiver operating characteristic curve for PET: A(PET) = 0.96 +/- 0.02 was significantly greater (P < 0.001) than that observed for CT: A(CT) = 0.77 +/- 0.06. The distribution of maximum standardized uptake value (SUVmax) showed that all negative lesions have SUVmax below 5.0 whereas 75% of positive lesions were above 5.0 pointing to the fact that disease positivity is more likely in lesions with high SUV values.Conclusion: The results of this study confirm that whole-body FDG-PET is more accurate than conventional CT in the staging of patients with suspected recurrent colorectal carcinoma. PMID- 11008101 TI - 18F-FDG Uptake in the Anterior Mediastinum. Physiologic Thymic Uptake or Disease? AB - Purpose: We reviewed 18F-fluorodeoxyglucose (18F-FDG) studies to determine the frequency of thymic visualization, to characterize this uptake to facilitate differentiation from disease, and to ascertain effects of therapy on visualization.Methods: Hybrid positron emission tomography images performed on 14 patients with known or suspected malignancy before therapy, and on six of the patients after treatment, were reviewed. Mediastinal uptake was characterized by location, contour and intensity.Results: Thymic uptake seen in five patients, 13 16-years-old, was characterized by an anterior midline location, regular contours, and a mean uptake ration of 2.5, and involved large glands. In five patients with mediastinal disease, 16-23-years-old, uptake was more posterior and eccentric in location with irregular borders and a mean uptake ratio of 4.3. One patient had anterior (thymic) and posterior (disease) mediastinal uptake. Three patients, 15-22-years-old, without disease and small thymus glands had no uptake. Five patients, including two with baseline uptake, showed no thymic uptake subsequently. One patient, negative initially, had thymic uptake five months after therapy.Conclusions: Thymic 18F-FDG uptake occurs in younger patients, before or after treatment, and is associated with larger glands. Its midline anterior mediastinal location and mild intensity should facilitate discrimination from disease. PMID- 11008102 TI - Occupational Exposure in Nuclear Medicine and PET. AB - Purpose: With the increasing use of 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) for scanning in oncology in our center, a radiation dose survey was performed to determine the impact on staff exposure. Conventional nuclear medicine procedures such as gallium scan, bone scans, and sestamibi cardiac scans are used for comparative purposes.Procedure: Patients were measured using a hand-held radiation monitor (Victoreen 450-P) at various distances and times that replicate typical patient contact scenarios in the Diagnostic Imaging Department.Results: We present our findings from the survey and the implications these have on staff radiation exposure. The data suggest that emerging oncologic techniques such as PET, high dose gallium-67, and high dose Tl-201 do not represent a significantly greater occupational radiation hazard than conventional nuclear medicine procedures. PMID- 11008103 TI - Mice unresponsive to GM-CSF are unexpectedly resistant to cutaneous Leishmania major infection. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown to play a protective role in leishmanial infection. Mice with a null mutation in the gene for the beta common (beta c) chain of the receptors for GM-CSF, interleukin(IL)-3 and IL-5 (beta c-null mice) display normal steady state hemopoiesis and develop lung disease similar to the human condition, alveolar proteinosis, due to a lack of signaling by GM-CSF. We therefore expected to observe a heightened sensitivity to Leishmania major in the beta c-null mice. Surprisingly, the beta c-null mice were more resistant to cutaneous infection than wild-type (wt) mice. Upon intradermal injection of L. major promastigotes, fewer beta c-null mice developed cutaneous lesions than wt mice and these lesions were smaller and healed more rapidly than in wt mice. This resistance to disease was associated with a reduced percentage of in vitro infected beta c-null macrophages. Macrophages from beta c null mice displayed a more activated phenotype and produced increased amounts of nitric oxide following infection with L. major, both in vivo and in vitro. Paradoxically, however, the parasite burden in the draining lymph nodes was similar in both beta c-null and wt mice, suggesting that at least a subpopulation of cells was susceptible to the parasite. The mechanism preventing normal lesion development remains to be elucidated. PMID- 11008104 TI - Passage of human T-cell leukemia virus type-1 during progression to cutaneous T cell lymphoma results in myelopathic disease in an HTLV-1 infection model. AB - Studies comparing functional differences in human T-cell leukemia virus type 1 (HTLV-1) clones that mediate distinct outcomes in experimentally infected rabbits, resulted in a dermatopathic smoldering adult T-cell leukemia/lymphoma following chronic infection with HTLV-1 strain RH/K34. During the 3.5 years' follow-up, HTLV-1 skin disease progressed to cutaneous T-cell lymphoma. When infection was passed to several naive rabbits, progressive paraparesis due to myelopathic neurodegeneration, analogous to HTLV-associated myelopathy, resulted in one of 4 transfusion recipients. Similar proviral loads were detected in the two diseases, regardless of stage of progression or tissue compartment of infection. Complete proviral sequences obtained from the donor and affected recipient aligned identically with each other and with the inoculated virus clone. Existence of disparate pathogenic outcomes following infectious transmission further extends the analogy of using rabbits to model human infection and disease. Although the experimental outcomes shown are limited by numbers of animals affected, they mimic the infrequency of HTLV-1 disease and authenticate epidemiological evidence of virus sequence stability regardless of disease phenotype. The findings suggest that further investigation of a possible role for HTLV-1 in some forms of cutaneous T-cell lymphoma is warranted. PMID- 11008105 TI - Lipopolysaccharide-like molecules derived from Wolbachia endobacteria of the filaria Onchocerca volvulus are candidate mediators in the sequence of inflammatory and antiinflammatory responses of human monocytes. AB - The majority of Onchocerca volvulus-infected persons show signs of cellular anergy, and long-time survival of adult and larval parasites in subcutaneous tissue is observed. The mechanisms leading to immunological hyporesponsiveness are poorly understood. Monocytes/macrophages represent a link between the innate and acquired immune system and are candidate cells to promote inflammatory and antiinflammatory processes. In the present study we have shown that products of microfilarial (O. volvulus) and adult (O. volvulus and O. ochengi) parasites affect monocytes in vitro. An early production of TNF-alpha by exposed monocytes was followed by the production of IL-10 and a reduced expression of HLA-DR and the costimulatory molecules B7-1 and B7-2, while other adhesion receptors remained unaffected. Downregulation of the functional membrane receptors failed to occur after treatment of the cells with anti-IL-10 antibodies. The engagement of CD14, a dominant membrane receptor on monocytes and major binding protein for lipopolysaccharides, was indicated by partial blocking of monocyte modulation by neutralizing antibodies to CD14 and by the antagonistic lipid A analog compound 406. Lipopolysaccharide-like molecules were detected in sterile products of O. volvulus stages which could originate from Wolbachia bacteria related to Gram negative Rickettsiales, known to be abundant in the hypodermis and the female reproductive organs of O. volvulus. The present results indicate that the monocyte/macrophage may be a major target cell for immunomodulatory parasite derived and intraparasitic, bacteria-derived molecules, thereby contributing to the host's cellular hyporesponsiveness. PMID- 11008106 TI - Helicobacter pylori inhibits gastric cell cycle progression. AB - Helicobacter pylori infection of the gastric mucosa is associated with changes in gastric epithelial cell proliferation. In vitro studies have shown that exposure to H. pylori inhibits proliferation of gastric cells. This study sought to investigate the cell cycle progression of gastric epithelial cell lines in the presence and absence of H. pylori. Unsynchronized and synchronized gastric epithelial cell lines AGS and KatoIII were exposed to H. pylori over a 24-h period. Cell cycle progression was determined by flow cytometry using propidium iodide (PI), and by analysis of cyclin E, p21, and p53 protein expression using Western blots. In the absence of H. pylori 40, 45, and 15% of unsynchronized AGS cells were in G(0)-G(1), S, and G(2)-M phases, respectively, by flow cytometry analysis. When AGS cells were cultured in the presence of H. pylori, the S phase decreased 10% and the G(0)-G(1) phase increased 17% after 24 h compared with the controls. KatoIII cells, which have a deleted p53 gene, showed little or no response to H. pylori. When G1/S synchronized AGS cells were incubated with media containing H. pylori, the G(1) phase increased significantly (25%, P < 0.05) compared with controls after 24 h. In contrast, the control cells were able to pass through S phase. The inhibitory effects of H. pylori on the cell cycle of AGS cells were associated with a significant increase in p53 and p21 expression after 24 h. The expression of cyclin E was downregulated in AGS cells following exposure of AGS cells to H. pylori for 24 h. This study shows that H. pylori induced growth inhibition in vitro is predominantly at the G(0)-G(1) checkpoint. Our results suggest that p53 may be important in H. pylori-induced cell cycle arrest. These results support a role for cyclin-dependent kinase inhibitors in the G(1) cell cycle arrest exerted by H. pylori and its involvement in changing the regulatory proteins, p53, p21, and cyclin E in the cell cycle. PMID- 11008107 TI - Mechanisms of disposal of bacterial lipopolysaccharides by animal hosts. AB - Much of the very extensive literature describing the (bio)chemistry and biology of bacterial lipopolysaccharides (LPS, endotoxin) has dealt with the properties of these molecules as potent triggers of host responses. This brief review will focus on what has been learned recently about mechanisms by which the host can dispose of LPS and counter its often excessive stimulatory effects. PMID- 11008108 TI - The role of immune responses in bone loss during periodontal disease. AB - A network of cytokines and other soluble mediators unites the immune system and bone; bacterial infections induce immune responses which may perturb this network. Periodontal diseases are Gram-negative infections resulting in bone loss in the jaw. Evidence is presented that immune responses to these infections produces net resorption of bone. PMID- 11008109 TI - Bartonella henselae, B. quintana, and B. bacilliformis: historical pathogens of emerging significance. AB - Bartonella species were virtually unrecognized as modern pathogens of humans until the last decade. However, identification of Bartonella species as the agents of cat-scratch disease, bacillary angiomatosis, urban trench fever, and possible novel presentations of Carrion's disease has left little doubt of the emerging medical importance of this genus of organisms. The three primary human pathogenic bartonellae, Bartonella bacilliformis (Carrion's disease), B. henselae (cat-scratch disease), and B. quintana (trench fever), present noteworthy comparisons in the epidemiology, natural history, pathology, and host-microbe interaction that this review will briefly explore. PMID- 11008110 TI - Helicobacter pylori motility. AB - Motility is essential for Helicobacter pylori colonization. This review discusses the biochemistry, genetics and genomics of the H. pylori flagellum, and compares these features with well-characterized bacteria. PMID- 11008111 TI - Using genetics to probe host-virus interactions; the mouse mammary tumor virus model. AB - It is clear that there is genetic variation among different individuals in their susceptibility to infection by viruses and other pathogens. Identification of the genes involved in conferring resistance or susceptibility to viral infection will allow us to understand both mechanisms of infection and pathogenesis and to develop reagents for treating or preventing them. Because of the large number of genetically well-characterized inbred mouse strains and the ability to generate targeted germ line mutations, this species is particularly well-suited for such analysis. This review focuses on how the use of genetics to study the retrovirus mouse mammary tumor virus allowed the dissection of both the viral infection pathway and the response of the host to this infection. PMID- 11008112 TI - Version 2000: the new beta-lactamases of Gram-negative bacteria at the dawn of the new millennium. AB - beta-lactamases of Gram-negative bacteria are evolving dynamically. New developments include the production of enzymes with novel substrate profiles, reduced susceptibility to beta-lactamase inhibitors, and the simultaneous production of multiple types of beta-lactamases. The changes represent evolutionary upgrades which provide modern pathogens with a greater potential to resist beta-lactam antibiotics and cause formidable therapeutic, infection control, and diagnostic challenges. This review is a clinically oriented outline of recent developments in the beta-lactamase production of Gram-negative bacteria. PMID- 11008113 TI - Bacterial penetration across the blood-brain barrier during the development of neonatal meningitis. AB - Bacterial pathogens may breach the blood-brain barrier (BBB) and invade the central nervous system through paracellular and/or transcellular mechanisms. Transcellular penetration, e.g., transcytosis across the BBB has been demonstrated for Escherichia coli K1, group B streptococcus, Listeria monocytogenes, Citrobacter freundii and Streptococcus pneumonia strains. Genes contributing to invasion of brain microvascular endothelial cells include E. coli K1 genes ompA, ibeA, ibeB, and yijP. Understanding the mechanisms of bacterial penetration across the BBB may help develop novel approaches to preventing bacterial meningitis. PMID- 11008114 TI - Expression and role of superoxide dismutases (SOD) in pathogenic bacteria. AB - This review will be limited to the expression and roles of the family of metalloenzymes superoxide dismutases in pathogenic bacteria. Only animal pathogens will be described, with particular emphasis on those causing disease in man. PMID- 11008115 TI - Interleukin-18 (IL-18) and infectious diseases, with special emphasis on diseases induced by intracellular pathogens. AB - Interleukin-18 (IL-18) is a novel cytokine mainly produced by activated macrophages. IL-18 was originally called interferon-gamma inducing factor, due to its action in inducing IFN-gamma secretion from Th1 cells, NK cells and NKT cells. It has been reported that IL-18 may play important roles in various diseases including cancer and infectious diseases. This review deals with the roles of IL-18 in infectious diseases, with special emphasis on IL-18 in infectious diseases caused by intracellular pathogens including Mycobacterium tuberculosis, Mycobacterium leprae, Listeria monocytogenes and Salmonella typhimurium. PMID- 11008116 TI - Overview of the epidemiology, microbiology, and pathogenesis of Leptospira spp. in humans. AB - Leptospirosis is probably the world's most widespread zoonosis. It remains underdiagnosed largely due to the broad spectrum of signs and symptoms attributable to this spirochetal pathogen. Leptospira spp. cause a diversity of diseases from flu-like illness to Weil's syndrome with multi-organ failure. Recent epidemics may herald a change in virulence or an alteration in the balance between humans and their interactions with other host species and the environment. PMID- 11008117 TI - Structure and function of clostridial phospholipases C. AB - A range of clostridial species produce phospholipases C. The zinc metallo phospholipases C have related sequences but different properties. All of these enzymes may be arranged, like alpha-toxin as two-domain proteins. Differences in enzymatic, haemolytic and toxic properties may be explained by differences in amino acids at key positions. PMID- 11008118 TI - The divergent role of tumor necrosis factor receptors in infectious diseases. AB - Tumor necrosis factor (TNF) receptor types 1 and 2 are broadly expressed by most cell types and are activated by binding of either TNF or lymphotoxin-beta. TNF receptor-mediated immune reactions are critically important in the pathogenesis and control of a variety of infections caused by bacteria, viruses, protozoa, and fungi. This review summarizes recent findings on the role of TNF receptors in infectious diseases and discusses the divergent functions of these receptors in immune responses. PMID- 11008119 TI - Structure-based drug design of non-nucleoside inhibitors for wild-type and drug resistant HIV reverse transcriptase. AB - The generation of anti-HIV agents using structure-based drug design methods has yielded a number of promising non-nucleoside inhibitors (NNIs) of HIV reverse transcriptase (RT). Recent successes in identifying potent NNIs are reviewed with an emphasis on the recent trend of utilizing a computer model of HIV RT to identify space in the NNI binding pocket that can be exploited by carefully chosen functional groups predicted to interact favorably with binding pocket residues. The NNI binding pocket model was used to design potent NNIs against both wild-type RT and drug-resistant RT mutants. Molecular modeling and score functions were used to analyze how drug-resistant mutations would change the RT binding pocket shape, volume, and chemical make-up, and how these changes could affect inhibitor binding. Modeling studies revealed that for an NNI of HIV RT to be active against RT mutants such as the especially problematic Y181C RT mutant, the following features are required: (a) the inhibitor should be highly potent against wild-type RT and therefore capable of tolerating a considerable activity loss against RT mutants (i.e. a picomolar-level inhibitor against wild-type RT may still be effective against RT mutants at nanomolar concentrations), (b) the inhibitor should maximize the occupancy in the Wing 2 region of the NNI binding site of RT, and (c) the inhibitor should contain functional groups that provide favorable chemical interactions with Wing 2 residues of wild-type as well as mutant RT. Our rationally designed NNI compounds HI-236, HI-240, HI-244, HI-253, HI-443, and HI-445 combine these three features and outperform other anti-HIV agents examined. PMID- 11008120 TI - The stimulatory effects of cationic amphiphilic drugs on human platelets treated with thrombin. AB - The actions of eight cationic amphiphilic drugs on human platelets displayed three different effects according to drug concentration ranges. At lower concentrations (below approximately 25 microM), the drugs stimulated secretory responses induced by 0.2 U/mL of thrombin, while at concentrations in the 25-50 microM range they inhibited these responses. Above 50-100 microM, the drugs caused permeabilization of the platelet plasma membrane as measured by leakage of cytoplasmic adenine nucleotides. The effects of these agents on phosphoinositide metabolism were monitored in platelets prelabeled with (32)P-inorganic phosphate, such that phosphatidic acid (PA), phosphatidylinositol 4-phosphate (PIP), and phosphatidylinositol 4,5-bisphosphate (PIP(2)), but not phosphatidylinositol (PI), were labeled to equilibrium. In unstimulated platelets, the level of labeled PA decreased slightly (about 25%), with corresponding increases in PIP(2) labeling up to drug concentrations of about 50 microM. In contrast to the relatively small changes in PI and PIP(2), the levels of labeled PIP, precursor to PIP(2), increased 2- to 4-fold in both resting and thrombin-treated platelets from 5 microM up to about 50-100 microM of drugs and remained elevated throughout the permeabilization concentrations. [(32)P]PA increased 20-fold over control upon thrombin activation and 5-30 microM of drugs caused [(32)P]PA to increase 30 37 times over that seen in control, resting platelets; the concentration of drugs that potentiated thrombin-induced [(32)P]PA elevation corresponded to that causing the potentiation of platelet secretion. Higher drug concentrations decreased [(32)P]PA elevation. [(32)P]PIP(2) levels increased about 25% in response to thrombin treatment alone; low concentrations of drugs led to another 25% elevation. A significant decrease in [(32)P]PIP(2) was seen above 30 microM, corresponding to inhibition of platelet secretion. Concentrations of 5-30 microM of several psychoactive agents, both neuroleptics and antidepressants, potentiated the thrombin-induced activation of platelets as measured by dense granule secretion and increased turnover of phosphoinositides. Remarkably, all of the drugs increased the levels of PIP even in resting platelets, indicating that they have common effects apart from the specific receptor interactions currently attributed to them. These common effects, e.g. an increase in membrane fluidity such as is known to be caused by amphipathic agents, may be in part responsible for the observed overlapping psychotropic effects of tricyclic antidepressants and phenothiazines. PMID- 11008121 TI - Differential effects of fluoxetine on murine B-cell proliferation depending on the biochemical pathways triggered by distinct mitogens. AB - The effect of fluoxetine on mitogen-induced B-cell proliferation was studied. In particular, we analyzed the influence of fluoxetine on the signal transduction pathways triggered after stimulation with lipopolysaccharide (LPS) and anti immunoglobulin M antibodies (anti-IgM). We showed that fluoxetine had a dual effect on anti-IgM-stimulated B-cell proliferation: at optimal anti-IgM concentration, fluoxetine inhibited proliferation, whereas at suboptimal anti-IgM concentration, the drug enhanced proliferation. Fluoxetine exerted only an inhibitory effect on LPS-induced B-cell proliferation. Calcium influx seemed to be involved in these effects. PMID- 11008122 TI - Profiles of antioxidant/electrophile response element (ARE/EpRE) nuclear protein binding and c-Ha-ras transactivation in vascular smooth muscle cells treated with oxidative metabolites of benzo[a]pyrene. AB - Activation of nuclear protein binding to the antioxidant/electrophile response element (ARE/EpRE) by benzo[a]pyrene (BaP) in vascular smooth muscle cells (vSMCs) is associated with transcriptional deregulation of c-Ha-ras. This response may be mediated by oxidative intermediates of BaP generated during the course of cellular metabolism. To test this hypothesis, the profile of ARE/EpRE protein binding and transactivation elicited by BaP was compared with that of 3 hydroxy BaP (3-OH BaP) (0.03 to 3.0 microM), BaP 7,8-dihydrodiol (BaP 7,8-diol) (0.03 to 3.0 microM), BaP 3,6-quinone (BaP 3,6-Q) (0.0003 to 3.0 microM), and H(2)O(2) (25 to 100 microM). Specific protein binding to the consensus c-Ha-ras ARE/EpRE was observed in vSMCs treated with all BaP metabolites at concentrations considerably lower than those required for the parent compound. H(2)O(2), a by product of BaP 3,6-Q redox cycling, also increased binding to the ARE/EpRE. Treatment of vSMCs with oxidative BaP metabolites or H(2)O(2) transactivated the c-Ha-ras promoter in all instances, but the response was consistently half of the maximal induction elicited by BaP. Similar proteins cross-linked specifically to the consensus c-Ha-ras ARE/EpRE sequence in cells treated with BaP or its oxidative intermediates. The protein binding profile in the c-Ha-ras promoter was similar to that in the NADPH:quinone reductase gene (NQO(1)) and the glutathione S-transferase Ya gene (GSTYa) promoters, but the relative abundance of individual complexes was promoter-specific. We conclude that oxidative intermediates of BaP mediate activation of nuclear protein binding to ARE/EpRE and contribute to transcriptional de-regulation of c-Ha-ras in vSMCs. PMID- 11008123 TI - Peroxide-like oxidant response in lungs of newborn guinea pigs following the parenteral infusion of a multivitamin preparation. AB - The multivitamin solution is a major component responsible for the photo-induced generation of peroxides in parenteral nutrition. The lung is a target of oxidant injury; however, the specific role of infused peroxides is unknown. The aim of this study was to determine if parenteral multivitamins induce in the lung an oxidant challenge similar to that of peroxides. Newborn guinea pigs were infused with dextrose plus relevant concentrations of H(2)O(2) (0,250,500 microM) or multivitamins (0,1%), as well as parenteral nutrition supplemented with multivitamins (0,1%). After 4 days, total glutathione, glutathione-related enzymes, and oxidant-sensitive eicosanoids were measured in the lungs. Peroxides as well as multivitamins led to a significant decrease in glutathione and the activity of glutathione synthase, indicating that infused peroxides were not entirely transformed into free radicals, which would have stimulated glutathione synthesis. The multivitamin solution induced a response in oxidant-sensitive eicosanoids similar to the response to peroxides, suggesting an oxidant stress that was not alleviated by the antiradical properties of its components. The effects on prostaglandins occurred independently from the stimulation in glutathione levels induced by parenteral nutrition. The multivitamin solution carries an oxidant load and causes effects similar to those of peroxides in the lungs of newborn guinea pigs. PMID- 11008124 TI - Increased nucleotide excision repair in cisplatin-resistant ovarian cancer cells: role of ERCC1-XPF. AB - Increased platinum-DNA adduct removal has been shown by several DNA repair assays to be associated with cisplatin resistance in the A2780/C-series human ovarian cancer model system. In the present study, we provide further evidence that the resistance phenotype of these cell lines is due, in part, to enhanced nucleotide excision repair (NER). Cisplatin resistance was found to be associated with increased UV resistance. Northern blot analysis revealed that increased expression of ERCC1 was also associated with cisplatin resistance in this panel. Several other NER genes were found to be constitutively overexpressed in the most resistant cell line, C200, as compared with the parental A2780 cells. A plasmid substrate containing a site-specific cisplatin adduct was used to measure the nucleotide excision activity of cell extracts prepared from cisplatin-sensitive and -resistant cells. Using this in vitro assay, extracts prepared from C200 cells exhibited approximately 3-fold more activity than extracts prepared from A2780 cells, similar to the difference in UV sensitivity. Complementation of A2780 extracts with ERCC1-XPF protein resulted in approximately 2-fold increased activity, but had little effect on excision in C200 extracts. Overall, these results support a role for the ERCC1-XPF endonuclease as a determinant of increased NER in this cisplatin resistance model. PMID- 11008125 TI - Inverse agonist properties of N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2, 4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl (SR141716A) and 1-(2 chlorophenyl)-4-cyano-5-(4-methoxyphenyl)-1H-pyrazole-3-carboxyl ic acid phenylamide (CP-272871) for the CB(1) cannabinoid receptor. AB - Two subtypes of cannabinoid receptors are currently recognized, CB(1), found in brain and neuronal cells, and CB(2), found in spleen and immune cells. We have characterized 1-(2-chlorophenyl)-4-cyano-5-(4-methoxyphenyl)-1H-pyrazole-3 carboxyl ic acid phenylamide (CP-272871) as a novel aryl pyrazole antagonist for the CB(1) receptor. CP-272871 competed for binding of the cannabinoid agonist (3)H-labeled (-)-3-[2-hydroxy-4-(1, 1-dimethylheptyl)-phenyl]-4-[3 hydroxypropyl]cyclohexan-1-ol ([(3)H]CP-55940) at the CB(1) receptor in rat brain membranes with a K(d) value 20-fold greater than that of N-(piperidin-1-yl)-5-(4 chlorophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide HCl (SR141716A). CP-272871 also competed for binding with the aminoalkylindole agonist (3)H-labeled (R)-(+)-[2, 3-dihydro-5-methyl-3-[(4 morpholinyl)methyl]pyrrolo[1,2,3-de]1, 4-benzoxazin-6-yl](1-naphthyl)methanone ([(3)H]WIN-55212-2), as well as the aryl pyrazole antagonist [(3)H]SR141716A. Inverse agonist as well as antagonist properties were observed for both SR141716A and CP-272871 in signal transduction assays in biological preparations in which the CB(1) receptor is endogenously expressed. SR141716A augmented secretin stimulated cyclic AMP (cAMP) accumulation in intact N18TG2 neuroblastoma cells, and this response was reversed by the agonist desacetyllevonantradol. CP-272871 antagonized desacetyllevonantradol-mediated inhibition of adenylyl cyclase in N18TG2 membranes, and increased adenylyl cyclase activity in the absence of agonist. SR141716A and CP-272871 antagonized desacetyllevonantradol-stimulated (35)S-labeled guanosine-5'-O-(gamma-thio)-triphosphate ([(35)S]GTPgammaS) binding to brain membrane G-proteins, and decreased basal [(35)S]GTPgammaS binding to G proteins. K(+) enhanced CP-272871 and SR141716A inverse agonist activity compared with Na(+) or NMDG(+) in the assay. These results demonstrated that the aryl pyrazoles SR141716A and CP-272871 behave as antagonists and as inverse agonists in G-protein-mediated signal transduction in preparations of endogenously expressed CB(1) receptors. PMID- 11008126 TI - CYP3A-inducing agents and the attenuation of uroporphyrin accumulation and excretion in a rat model of porphyria cutanea tarda. AB - An experimental model of porphyria cutanea tarda (PCT) can be achieved in 3 weeks by a single injection of a mixture of polychlorinated biphenyls (Aroclor 1254) into iron-loaded female Fischer 344 rats maintained continuously on delta aminolevulinic acid-supplemented drinking water. In this model, daily treatment with 5-pregnen-3 beta-ol-20-one-16 alpha-carbonitrile (pregnenolone 16 alpha carbonitrile) attenuated uroporphyrin and heptacarboxylporphyrin accumulation and excretion by 75%. Pregnenolone 16 alpha-carbonitrile treatment had only a minor effect on hepatic iron stores, and it had no effect on the induction of CYP1A activities by Aroclor 1254. In the absence of Aroclor 1254, pregnenolone 16 alpha carbonitrile had no effect on the accumulation and excretion of highly carboxylated porphyrins. Attenuation of porphyrin accumulation could also be demonstrated with daily troleandomycin treatment. Troleandomycin increased CYP3A dependent erythromycin demethylase activity, but to a lesser extent than pregnenolone 16 alpha-carbonitrile. Much of the CYP3A induced by troleandomycin was sequestered as a catalytically inactive metabolic-intermediate complex. In the absence of Aroclor 1254, troleandomycin had no effect on the accumulation and excretion of highly carboxylated porphyrins, nor did troleandomycin alter the induction of CYP1A by Aroclor 1254. The results suggest that the major attenuation of hepatic accumulation and urinary excretion of uro- and heptacarboxylporphyrins in the rat PCT model by pregnenolone 16 alpha carbonitrile and troleandomycin is due to an enhancement of CYP3A catalytic activity. PMID- 11008127 TI - Characterization and effects of methyl-2- (4-aminophenyl)-1, 2-dihydro-1-oxo-7- (2-pyridinylmethoxy)-4-(3,4, 5-trimethoxyphenyl)-3-isoquinoline carboxylate sulfate (T-1032), a novel potent inhibitor of cGMP-binding cGMP-specific phosphodiesterase (PDE5). AB - An isoquinolone derivative, methyl-2-(4-aminophenyl)-1, 2-dihydro-1-oxo-7-(2 pyridinylmethoxy)-4-(3,4, 5-trimethoxyphenyl)-3-isoquinoline carboxylate sulfate (T-1032), was found to be a novel potent inhibitor of cyclic GMP (cGMP)-binding cGMP-specific phosphodiesterase (PDE5). We investigated the inhibitory effects of T-1032 on six PDE isozymes isolated from canine tissues. T-1032 specifically inhibited the hydrolysis of cGMP by PDE5 partially purified from canine lung, at a low concentration (IC(50) = 1.0 nM, K(i) = 1.2 nM), in a competitive manner. In contrast, the IC(50) values of T-1032 for PDE1, PDE2, PDE3, and PDE4 were more than 1 microM. T-1032 also inhibited PDE6 from canine retina with an IC(50) of 28 nM, which is of the same order of magnitude as the IC(50) of sildenafil. cGMP hydrolytic activities of two alternative splice variants of canine PDE5 expressed in COS-7 cells were inhibited by this compound to a similar extent. T-1032 increased the intracellular concentration of cGMP in cultured rat vascular smooth muscle cells in the presence and absence of C-type natriuretic peptide, an activator of membrane-bound guanylate cyclase, whereas the compound did not change cyclic AMP levels. These data indicated that T-1032, which belongs to a new structural class of PDE5 inhibitors, is a potent and selective PDE5 inhibitor. This compound may be useful in pharmacological studies to examine the role of a cGMP/PDE5 pathway in tissues. PMID- 11008128 TI - Prevention of cisplatin-DNA adduct repair and potentiation of cisplatin-induced apoptosis in ovarian carcinoma cells by proteasome inhibitors. AB - Histones H2A and H2B are known to be reversibly post-translationally modified by ubiquitination. We previously observed in cultured tumor cells that proteasome inhibition stabilizes polyubiquitinated proteins, depletes unconjugated ubiquitin, and thereby promotes the deubiquitination of nucleosomal histones in chromatin. Provocative indirect evidence suggests that histone ubiquitination/deubiquitination cycles alter chromatin structure, which may limit accessibility of DNA repair proteins to damaged sites. In the present study, we focused on the relationship between the ubiquitination status of histone H2A, the structure of chromatin, and the efficiency of nucleotide excision repair (NER) of cisplatin-DNA adducts in human ovarian carcinoma cells exposed to the antitumor drug cisplatin. Pretreating cells with the proteasome inhibitor lactacystin (LC) or N-acetyl-leucyl-leucyl-norleucinal (ALLnL) induced deubiquitination of ubiquitinated histone H2A (uH2A) and concomitantly promoted chromatin condensation, increased the extent of cisplatin-DNA adducts, and diminished NER dependent repair of cisplatin-DNA lesions, compared with control cells treated with cisplatin alone. Both proteasome inhibitors also prevented the increase in ERCC-1 mRNA expression that occurs in cells exposed to cisplatin. Cells treated with the combination of ALLnL and cisplatin underwent apoptosis, as indicated by caspase-dependent poly(ADP-ribose) polymerase (PARP) cleavage, more quickly than cells treated with either agent alone. Additionally, the combination of ALLnL and cisplatin potently increased p53 levels in cell lysates and stimulated the binding of p53 to chromatin. Together, these observations suggest that proteasome inhibition may be exploited therapeutically for its potential to sensitize ovarian tumor cells to cisplatin. PMID- 11008129 TI - Effects of resveratrol on the autophosphorylation of phorbol ester-responsive protein kinases: inhibition of protein kinase D but not protein kinase C isozyme autophosphorylation. AB - The natural product resveratrol is a potent antagonist of phorbol ester-mediated tumor promotion and in vitro cellular responses to phorbol-ester tumor promoters, but it is only weakly inhibitory against the phosphorylation of conventional exogenous substrates by phorbol ester-responsive protein kinase C (PKC) isozymes. In this report, we compare the effects of resveratrol against the autophosphorylation reactions of PKC isozymes versus the novel phorbol ester responsive kinase, protein kinase D (PKD). We found that resveratrol inhibits PKD autophosphorylation in a concentration-dependent manner, but has only negligible effects against the autophosphorylation reactions of representative members of each PKC isozyme subfamily (cPKC-alpha, -beta(1), and -gamma, nPKC-delta and epsilon, and aPKC-zeta). Resveratrol was comparably effective against PKD autophosphorylation (IC(50) = 52 microM) and PKD phosphorylation of the exogenous substrate syntide-2 (IC(50) = 36 microM). The inhibitory potency of resveratrol against PKD is in line with the potency of resveratrol observed in cellular systems and with its potency against other purified enzymes and binding proteins that are implicated in the cancer chemopreventive activity of the polyphenol. Thus, PKD inhibition may contribute to the cancer chemopreventive action of resveratrol. PMID- 11008130 TI - Correlation of a unique 220-kDa protein with vitamin D sensitivity in glioma cells. AB - We have investigated the antitumor and apoptotic effects of 1, 25 dihydroxyvitamin D(3) (VD(3)) in glioma cell lines and in primary cultures derived from surgical specimens from patients. Our results showed that certain glioma cells underwent apoptosis, whereas others were resistant. In an attempt to search for parameters that dictate VD(3) sensitivity, we discovered a unique 220 kDa protein in glioma cells that were sensitive to VD(3). This protein was not a classical vitamin D receptor (VDR), but was recognized by two different anti-VDR monoclonal antibodies. Furthermore, the level of the 220-kDa protein was inversely correlated with the IC(50) of VD(3) in these glioma cells. This 220-kDa protein was also present in frozen brain tumor samples, and the level of expression appeared to correlate with their corresponding primary cultures. Thus, our findings suggest that this 220-kDa protein may play an important role in determining VD(3) sensitivity in malignant glioma. PMID- 11008131 TI - Novel mechanisms of DNA topoisomerase II inhibition by pyranonaphthoquinone derivatives-eleutherin, alpha lapachone, and beta lapachone. AB - Pyranonaphthoquinones have diverse biological activities against Gram-positive bacteria, fungi, and mycoplasms, and, recently, there has also been an increasing interest in their anti-cancer activity. This study includes three derivatives: eleutherin (compound 1), beta lapachone (compound 2), and its structural isomer, alpha lapachone (compound 3). The mechanism of topoisomerase II inhibition by the three derivatives was examined systematically with respect to the steps of the catalytic cycle of the enzyme. Etoposide, the prototypical enzyme poison, was used as a control and in combination with compounds 1-3 to localize their mechanism of action. The study revealed that eleutherin (1) and beta lapachone (2) inhibited topoisomerase II by inducing religation and dissociation of the enzyme from DNA in the presence of ATP. Whereas compound 2 was an "irreversible" inhibitor of topoisomerase II, compound 1 merely slowed the catalytic cycle of the enzyme. alpha Lapachone (3), on the other hand, inhibited initial non covalent binding of topoisomerase II to DNA and, in addition, induced religation of DNA breaks (even in pre-established ternary complexes) before dissociating the enzyme from DNA. Compound 3 was an "irreversible" inhibitor of topoisomerase II. The diverse and unique mechanisms of topoisomerase II inhibition by pyranonaphthoquinone derivatives reveal novel ways to target the enzyme with potential for anti-cancer drug design. PMID- 11008132 TI - Phospholipids as multidrug resistance modulators of the transport of epirubicin in human intestinal epithelial Caco-2 cell layers and everted gut sacs of rats. AB - Phospholipids have been increasingly used as carriers for the delivery of a variety of drugs. Studies using cancer chemotherapeutic agents such as epirubicin encapsulated in liposomes, which are made of phospholipids and other ingredients, have generally shown reduced toxicity and enhanced therapeutic efficacy. The recent investigation of the role of P-glycoprotein (P-gp) in phospholipid translocation has opened a new area of research on the possible use of phospholipids as multidrug resistance (MDR) modulators. This study investigated the effects of liposomal encapsulation, empty liposome pretreatment, or free lipid pretreatment on the uptake and transport of epirubicin in the human colon adenocarcinoma cell line Caco-2 and in everted gut sacs of rat jejunum and ileum. Epirubicin uptake experiments, using a flow cytometer, showed that both liposomal encapsulation and empty liposome pretreatment increased the intracellular accumulation of epirubicin in Caco-2 cells significantly. These two treatments substantially increased apical-to-basolateral absorption of epirubicin across Caco-2 monolayers and markedly improved mucosal-to-serosal absorption of epirubicin in rat jejunum and ileum. Enhancement also was observed with both liposome encapsulation and empty liposome pretreatment in the reduction of basolateral-to-apical efflux of epirubicin across Caco-2 monolayers. However, because diffusion of free dipalmitoyl phosphatidylcholine (DPPC) or dipalmitoyl phosphatidylethanolamine (DPPE) lipids across the cell membrane is very slow, these free lipids showed marginal effects on absorption and/or secretion of epirubicin in both Caco-2 cells and rat gut sacs. The study suggests that inhibition of P-gp or other transporter proteins located in the intestines may be partially involved in the reduction of epirubicin efflux. In conclusion, the therapeutic efficacy of epirubicin may be improved by using phospholipids as excipients and MDR modulators in the formulations. Liposomal formulations may have important applications to circumvent drug resistance in cancer chemotherapy. PMID- 11008133 TI - Promitogenic effects of ethanol, methanol, and ethanolamine in insulin-treated fibroblasts. AB - The zinc-dependent potentiating effect of ethanol (EtOH) on insulin-stimulated DNA synthesis was studied with a focus on the possible site of EtOH action and the ability of other alcohols to elicit similar promitogenic effects. In serum starved (27 hr) NIH 3T3 fibroblasts, 200-300 mM methanol (MeOH) and 0.1-1.5 mM ethanolamine (Etn), but not 3- to 9-carbon normal alcohols, enhanced the effect of insulin on DNA synthesis to varying extents. The promitogenic effects of EtOH and MeOH, but not that of Etn, required the presence of 15-25 microM zinc. The potentiating effects of Etn were enhanced by 5 mM choline (Cho) and inhibited by 1-3 mM hemicholinium-3 (HC-3), an inhibitor of Cho transporter and Cho kinase. In the presence of 15 microM zinc, 40 mM EtOH, which had no effect on its own, inhibited the potentiating effects of Cho and enhanced the inhibitory effects of HC-3 on synergistic stimulation of DNA synthesis by Etn and insulin. On the other hand, both Cho and HC-3 partially inhibited the promitogenic effect of 80 mM EtOH in the presence of 25 microM zinc. After a 10-min incubation, EtOH decreased the amount of cell-associated [(14)C]Cho in the absence but not in the presence of HC 3. After a 40-min incubation, Cho (5 mM) partially inhibited the cellular uptake as well as the metabolism of [(14)C]Etn. Whereas after the 40-min incubation 80 mM EtOH had no effects on Etn metabolism, in the absence of Cho it decreased the amount of cell-associated [(14)C]Etn. However, EtOH had no detectable effects on cell association of [(14)C]Etn after the 10-min incubation. The results suggest that in NIH 3T3 fibroblasts EtOH is a remarkably specific promitogen, and that it may act via a cell membrane site(s), also regulated by Cho (agonist) and HC-3 (antagonist), which can influence membrane binding and the promitogenic activity of Etn. PMID- 11008135 TI - What limits simultaneous discrimination accuracy? AB - Discrimination accuracy decreases when viewers simultaneously monitor two perceptually distinct stimulus components for changes in a common property, e.g. contrast [Magnussen & Greenlee (1997). Journal of Experimental Psychology: Human Perception and Performance, 23, 1603-1616; Olzak & Wickens (1997). Perception, 26, 1101-1120]. We ask whether the limitation is in monitoring two components or in making dual decisions about a single property. Using the same uncertainty paradigm as Magnussen and Greenlee, we find no evidence of a processing limitation when viewers simultaneously monitor one component (1.25 c/d) for a possible change in contrast and a second component (5 c/d) for a possible change in spatial frequency, regardless of whether the components are spatially separated or superimposed. The limitation is in making dual decisions about a single property. PMID- 11008134 TI - Inhibition of agonist-induced p42 and p38 mitogen-activated protein kinase phosphorylation and CD40 ligand/P-selectin expression by cyclic nucleotide regulated pathways in human platelets. AB - Platelet activation and adhesion to endothelial cells and extracellular matrix proteins are crucial events in the development of arterial cardiovascular diseases. Platelet activation is initiated by stimulation of intracellular signaling cascades, including the p42 mitogen-activated protein kinase (MAPK) and p38 MAPK pathways, followed by major changes in the platelet cytoskeleton and expression and activation of platelet surface receptors, such as P-selectin (CD62P) and CD40 ligand (CD40L). Activated platelets directly bind to vascular endothelial cells via CD40L/CD40 interactions and induce inflammatory reactions that initiate or aggravate atherosclerotic lesions. The aim of this study was to investigate effects of two known platelet inhibitors-the cAMP-elevating prostaglandin E(1) (PG-E(1)) and the cGMP-elevating sodium nitroprusside (SNP)-on platelet p42 MAPK and p38 MAPK activation as well as on surface expression of CD62P and CD40L. MAPK activation was analyzed by Western blot experiments using phosphorylation-specific antibodies, and surface CD40L and CD62P expression was determined by flow cytometry analysis. PG-E(1) and SNP strongly inhibited p42 and p38 MAPK phosphorylation as well as CD40L and CD62P expression in response to thrombin, a thromboxane A(2) analog, and ADP. These data indicate that adenosine and guanosine 3',5'-cyclic monophosphate-dependent protein kinases not only inhibit platelet pathways leading to activation and aggregation, but also those resulting in enhanced surface expression of protein ligands involved in inflammation. Expression of CD40L and CD62P was found to be independent of MAPK activation, since it was not inhibited by specific MAPK inhibitors. Inhibition of platelet-induced inflammatory responses including CD62P- and CD40L-mediated interaction of platelets with leukocytes and endothelial cells, respectively, is suggested to be an important component of the long-term vasoprotective effects of cyclic nucleotide-elevating prostaglandins and NO donors. PMID- 11008136 TI - A central mechanism of chromatic contrast. AB - The color appearance of a light can be altered by introducing a second, surrounding field. This phenomenon, called chromatic induction, is attenuated by chromatic variation within a remote region outside the surround [Shevell & Wei (1998). Vision Research, 38, 1561-1566]. We now consider the locus of the neural mechanism mediating the attenuation caused by the remote chromatic contrast. In the first experiment, the magnitude of chromatic variation within the remote region is changed either: (i) in the same eye that views the patch judged in color; or (ii) in only the opposite eye. The measurements are virtually the same in both cases, which implies attenuation of chromatic induction is mediated by a central, binocular mechanism. In the second experiment, the patch with its immediate inducing surround is changed in binocular disparity relative to the remote region with chromatic variation. The patch and surround, seen together in one depth plane, are perceived to be in front of, behind, or in the same plane as the remote region with chromatic variation. Attenuation of chromatic induction is strongest when the patch and surround are in the same depth plane as the remote region. This change of color appearance with disparity is consistent with a central binocular process. Overall, the color-appearance measurements are explained by monocular encoding of chromatic differences at edges, and a central binocular mechanism of chromatic-contrast gain control. PMID- 11008137 TI - Salience from feature contrast: variations with texture density. AB - The salience of popout targets was measured in regular line arrays as a function of texture density. Test targets (singletons with orientation, motion, or luminance contrast) presented at different raster widths were compared with reference lines (lines brighter than surrounding lines) presented at fixed raster width. The luminance at which the reference target appeared as salient as the particular test target was taken as a measure of the relative salience of the test target. For orientation or motion contrast, targets at medium to small raster widths were far more salient than targets in sparse or very dense line arrangements. For targets defined by luminance contrast, salience variations with texture density were less pronounced. Some subjects also reported salience for lines in sparse arrangements even when these did not display feature contrast. When such non-specific saliency effects were subtracted from the actual measurements, salience curves for orientation or motion contrast revealed peaks of increased sensitivity at line spacings below 2-3 deg and flat curves at larger grid sizes. In an additional experiment, saliency effects from orientation contrast were measured using texture lines of different size. Salience variations were commonly observed. However, the curves were not found to scale with the different sizes of texture elements but were constantly related to the free space between neighbouring lines. This suggests that peaks in the salience profiles reflect the limited spatial extent of the underlying neural mechanisms. PMID- 11008138 TI - Exposure duration affects the perceived direction of cyclopean type II plaids. AB - This study investigated the effect of exposure duration on the perceived direction of cyclopean Type I and Type II plaids moving in the X/Y plane. The cyclopean plaids were created from grating components defined by binocular disparity embedded in a dynamic random-dot stereogram. The results showed that the cyclopean Type I plaid appeared to move in the intersection-of-constraints (IOC) direction across the range of exposures tested. However, the cyclopean Type II plaids appeared to move in a direction different from the IOC with short exposures but near the IOC with long exposures. This perceived directional shift was also obtained with luminance-defined Type II plaids. A common pattern-motion mechanism that processes cyclopean and luminance motion signals appears responsible for the perceived directional shift of the Type II plaids. PMID- 11008139 TI - Perceptual learning for a pattern discrimination task. AB - Our goal was to differentiate low and mid level perceptual learning. We used a complex grating discrimination task that required observers to combine information across wide ranges of spatial frequency and orientation. Stimuli were 'wicker'-like textures containing two orthogonal signal components of 3 and 9 c/deg. Observers discriminated a 15% spatial frequency shift in these components. Stimuli also contained four noise components, separated from the signal components by at least 45 degrees of orientation or approximately 2 octaves in spatial frequency. In Experiment 1 naive observers were trained for eight sessions with a four-alternative same-different forced choice judgment with feedback. Observers showed significant learning, thresholds dropped to approximately 1/3 of their original value. In Experiment 2 we found that observers showed far less learning when the noise components were not present. Experiment 3 found, unlike many other studies, almost complete transfer of learning across orientation. The results of Experiments 2 and 3 suggest that, unlike many other perceptual learning studies, most learning in Experiment 1 occurs at mid to high levels of processing rather than within low level analyzers tuned for spatial frequency and orientation. Experiment 4 found that performance was more severely impaired by spatial frequency shifts in noise components of the same spatial frequency or orientation as the signal components (though there was significant variability between observers). This suggests that after training observers based their responses on mechanisms tuned for selective regions of Fourier space. Experiment 5 examined transfer of learning from a same-sign task (the two signal components both increased/decreased in spatial frequency) to an opposite-sign task (signal components shifted in opposite directions in frequency space). Transfer of learning from same-sign to opposite-sign tasks and vice versa was complete suggesting that observers combined information from the two signal components independently. PMID- 11008140 TI - Spatial color contrast matching: broad-bandpass functions and the flattening effect. AB - The contrast matching function (CMF) is the reciprocal of test contrast that perceptually matches the contrast of standard pattern, measured as a function of test spatial frequency (SF). Achromatic CMFs usually flatten as the contrast of the standard is raised, and are broader than the achromatic, bandpass, contrast sensitivity function (CSF). This report investigates whether chromatic CMFs have similar characteristics. For this purpose, the red-green color channel was defined using minimum flicker and hue cancellation techniques. Spatially localized (D6), vertical, equiluminant patterns (SFs: 0.063-8 cpd; contrast: 3 80%) were used to measure the CSF and CMF of isoluminant patterns presented with a temporal Gaussian envelope. CMFs were measured using a randomized double staircase procedure and the two-interval forced choice technique. Two color normal observers, whose task was to select the interval that had higher color contrast, participated in experiments. Results show that: (a) the color CMFs are lowpass functions of SF at low standard contrasts (3-12.5%), broad-bandpass at intermediate contrasts (6.25-60%), and near-flat at high contrasts (80%); and (b) isoluminant CMFs have higher upper cut-off frequencies than isoluminant CSFs. It is concluded that: (i) color-contrast-constancy (CMF independent of SF) is partly achieved at high contrasts because color CMFs flatten as contrast increases; (ii) the information processing at suprathreshold levels is different from that at the threshold levels; and (iii) the model that explained achromatic CMFs using achromatic threshold mechanisms could not explain chromatic CMFs using chromatic threshold mechanisms. PMID- 11008141 TI - Relative contributions of sustained and transient pathways to human stereoprocessing. AB - It has been proposed [Hubel & Livingstone (1987) Journal of Neuroscience, 7, 3378 3415] that stereopsis is mediated solely by magnocellular pathway in primates. This hypothesis was evaluated for humans in psychophysical experiments with dynamic random-noise stimuli, based on the sustained/transient relationship of behavior mediated by the two divisions of the LGN [Merigan & Maunsell (1993) Annual Review of Neuroscience, 16, 369-402]. The stereoscopic limits show that stereoscopic system is more sensitive to sustained random-dot stimuli than to transient ones. Quantitative modeling of the result implied a weak role for magnocellular input, suggests that human stereopsis is more strongly influenced by parvocellular input through the LGN. PMID- 11008142 TI - Spectral tuning of dichromats to natural scenes. AB - Multispectral images of natural scenes were collected from both forests and coral reefs. We varied the wavelength position of receptors in hypothetical dichromatic visual systems and, for each receptor pair estimated the percentage of discriminable points in natural scenes. The optimal spectral tuning predicted by this model results in photoreceptor pairs very like those of forest dwelling, dichromatic mammals and of coral reef fishes. Variations of the natural illuminants in forests have little or no effect on optimal spectral tuning, but variations of depth in coral reefs have moderate effects on the spectral placement of S and L cones. The ratio of S and L cones typically found in dichromatic mammals reduces the discriminability of forest scenes; in contrast, the typical ratio of S and L cones in coral reef fishes achieves nearly the optimal discrimination in coral reef scenes. PMID- 11008143 TI - Optical correction of form deprivation myopia inhibits refractive recovery in chick eyes with intact or sectioned optic nerves. AB - The finding that the eyes of young chicks recover quickly from form deprivation myopia (FDM) has been interpreted as indirect evidence for active emmetropization. More direct evidence would be the demonstration that correction of FDM with spectacle lenses, thereby removing the defocus signal, prevents recovery. We investigated this issue in eyes with intact and sectioned (ONS) optic nerves. Previous studies suggest that an intact optic nerve is necessary for accurate emmetropization. Seventy day-old male chicks were monocularly deprived using velcro-mounted diffusers, which were removed after 5-6 days and in some (n=51), but not all cases, replaced by spectacle lenses (-5, -10 or -15 D). Approximately half (n=34) of the chicks also underwent ONS on day 1. Refractive errors and axial ocular dimensions were measured when the diffusers were first removed and thereafter at 2-4 day intervals over the following 1-2 weeks. In one case, measurements were continued at less regular intervals to 33 days. Lens powers were selected to either approximately correct or under-correct the refractive errors present when the diffusers were removed. Form deprivation in normal chicks produced large myopic shifts in refraction (means for groups range from -9.20 to -16.07 D). When the deprivation treatment was terminated, the myopia quickly decreased to negligible levels unless optically corrected. Correcting lenses stabilized the myopia to a level consistent with the lens power used. Interocular differences in axial length were consistent with an axial origin to the refractive changes. Results for the ONS groups exhibited similar trends although there was increased variability in the data. The findings support the interpretation that recovery from FDM is a product of active emmetropization. That ONS increased the variability of such responses implies that an intact optic nerve is required for accurate emmetropization. PMID- 11008144 TI - Influence of monocular deprivation during infancy on the later development of spatial and temporal vision. AB - Using the method of limits, we measured spatial and temporal vision in 15 patients, aged 4-28 years, who had been monocularly deprived of patterned visual input during infancy by a dense cataract. All patients showed losses in both spatial and temporal vision, with greater losses in spatial than in temporal vision. Losses were smaller when there had been more patching of the non-deprived eye. The results indicate that visual deprivation has smaller effects on the neural mechanisms mediating temporal vision than on those mediating spatial vision. PMID- 11008146 TI - In this issue PMID- 11008145 TI - Amblyopic deficits in detecting a dotted line in noise. AB - We compared detectability of a dotted line masked by random-dot noise for the amblyopic versus non-amblyopic eye of two strabismic amblyopes. Small but consistent deficits in the amblyopic eye of these observers were found, and shown to be limited to dotted-line targets composed of greater than seven dots (with performance being normal for targets of less than seven dots). These deficits were unrelated to impaired visual acuity, impaired sensitivity to dot density, and differential positional uncertainty between the eyes of our observers. The deficits were also unlikely to be due to CSF losses due to abnormal low-spatial frequency filters involved in detecting long chains of collinear dots. Instead, the results of simulations indicate that the inefficiency in utilising large numbers of dots is due to deficits of global, integrative processes in strabismic amblyopes. These simulations also show that while neither undersampling nor positional uncertainty of inputs into integrative processes can themselves account for the amblyopic deficits, if such abnormal inputs lead to the development of stunted integrative processes then impaired sensitivity to long chains of collinear dots is indeed predicted. PMID- 11008147 TI - Harold Hillman MBBS MRCS LRCP BSc PhD. PMID- 11008148 TI - Aminophylline in the treatment of atropine-resistant bradyasystole. PMID- 11008149 TI - Anaesthesia and critical care considerations in nerve agent warfare trauma casualties. AB - Nerve agents (NA) (tabun, sarin, suman, VX) have been stocked around the world for some time and still present a major threat to civilian as well as to military populations. Since NA can be delivered through both an aerial spray system and a ballistic system, victims could suffer both NA intoxication and multiple trauma necessitating urgent surgical intervention followed by intensive care. These patients can be expected to be extremely precarious neurologically, respiratorily and haemodynamically. Moreover, their clinical signs can be misleading. Further exacerbating the problem is the fact that interactions of NA with the pharmacological agents used for resuscitation and/or during anaesthesia can aggravate organ instability even more and possibly cause systemic collapse. There are no protocols for perioperative critical care and early assessment or for the administration of anaesthesia for surgical interventions in such combined multiple trauma and intoxicated casualties. We propose a scheme for the administration of critical care and anaesthesia based on the scant anecdotal reports that have emerged after the occurrence of local accidents involving NA intoxication and on the neuropharmacological knowledge of the pesticide organophosphate poisoning database, these compounds being related chemical substances. PMID- 11008150 TI - Outcome after cardiac arrest in adults in UK hospitals: effect of the 1997 guidelines. AB - AIM: To assess the effectiveness of the ILCOR Advisory Statements on Advanced Life Support adopted by the Resuscitation Council (UK), as the standard for resuscitation following cardiac arrest. METHOD: Over the period May to November 1997, data on the process and outcome of cardiopulmonary resuscitation following in-hospital cardiac arrest were collected from 49 hospitals throughout the UK. RESULTS: Of 2074 audit forms submitted, 1368 were included in the final analysis. The initial rhythm monitored was ventricular fibrillation (VF) or pulseless ventricular tachycardia (VT) in 429 patients, of whom 181 (42.2%) were discharged alive, compared to 6. 2% when the initial rhythm was non-VF/VT. Overall, 240 (17.6%) patients were discharged alive. At 6 months after discharge 195 (82. 3%) of 237 patients were still alive. Successful initial resuscitation, defined as return of spontaneous circulation lasting longer than 20 min (ROSC>20 min), was significantly associated with VF/VT as the initial arrest rhythm, return of circulation in less than 3 min, age less than 70 years and the use of an advanced airway (P<0.01). There was a significant increased likelihood of survival to discharge when the circulation was restored in less than 3 min and age was less than 70 years (P<0.05). The administration of any adrenaline (epinephrine) was significantly associated with a reduced likelihood of ROSC>20 min or alive discharge (P<0.0001). CONCLUSION: Compared to the last major multiple hospital study published in 1992, the results of this study suggest that there appears to have been an improvement in survival of in-hospital patients in the UK who have a VF/VT cardiac arrest. How much of this is directly attributable to the adoption of the latest guidelines is uncertain. PMID- 11008151 TI - One-year survival after out-of-hospital cardiac arrest in Copenhagen according to the 'Utstein style'. AB - OBJECTIVE: To determine survival after out-of-hospital cardiac arrest (CA) in Copenhagen, according to the Utstein recommendations, and compare this with other emergency medical services systems. DESIGN: Register-based cohort study. SETTING: Copenhagen, population 465000, area 90 km(2). PATIENTS: Consecutive group of patients with out-of-hospital CA occurring between January 1 1991 and December 31 1993, followed up via the hospital database systems. MATERIALS: Two specially equipped advanced life support (ALS) units, staffed with an anaesthesiologist and a specially trained fireman, operating to support basic life support units. RESULTS: Of 2225 patients who were unconscious without a pulse or breathing, 1461 were declared dead by the anaesthesiologist. Advanced cardiac life support was initiated in 764, 61 of which were of non-cardiac aetiology. The presumed aetiology was cardiac in 703: in 235 the event was unwitnessed, in 464 witnessed and in four the information was missing. Of 464 witnessed CA the initial rhythm was asystole in 72 cases, in 302 ventricular fibrillation (VF) or ventricular tachycardia (VT), and in 90 were in other rhythms. In these subgroups discharged rates were 5 (7%), 62 (21%) and 1 (1%), and 1-year survival rates were 4 (6%), 49 (16%) and 1 (1%), respectively. The median ALS call-response interval was 6 min. CONCLUSIONS: Survival after CA is more likely if the collapse was witnessed and in patients with VF/VT of cardiac aetiology. PMID- 11008152 TI - The attitudes of cardiac arrest survivors and their family members towards CPR courses. AB - OBJECTIVES: to evaluate self-assessment of first aid knowledge, readiness to make use of it in case of a medical emergency and judgement of a 1-day CPR course by cardiac arrest survivors, their family members and friends as compared to the general public. BACKGROUND: the recurrence rate of a cardiac arrest after successful resuscitation is high and most of out-of-hospital cardiac arrests occur at the patient's home. METHODS: medical students trained in basic and advanced life support provided 101 members of the target group and 94 of a sex and age matched control group with a 1-day course in CPR. RESULTS: after the course, half of the participants in both groups considered their knowledge of first aid to be very good or good. The readiness to perform first aid in a medical emergency increased significantly. Of the target group 96% of the participants as compared with the control group where 91% felt confident to recognise a cardiac arrest; 79 versus 68% considered themselves capable to perform CPR if needed. The course was judged as very good in 71 versus 69% and as good in 25 versus 27% with no differences between groups. CONCLUSION: one-day CPR courses are well accepted by cardiac arrest survivors, their family members and friends and help to reduce fears of reacting in medical emergencies. They seem to be more motivated to gain and use first aid knowledge than others. PMID- 11008153 TI - Successful out-of-hospital cardiopulmonary resuscitation: what is the optimal in hospital treatment strategy? AB - The aim of the study was to evaluate prognostic factors in patients after successful out-of-hospital resuscitation (sOHR) within 30 min after admission. A prognostic scoring scale in patients surviving OHR was analysed. We also studied the effect of these predictive factors and the in-hospital treatment (percutaneous transluminal coronary angioplasty (PTCA) vs. thrombolysis) on mortality. We performed a retrospective analysis of the emergency medical system forms and medical files of 72 consecutive patients aged > or =18 years with sOHR. Of these 72 patients 37 (51%) met the electrocardiographic and enzymatic criteria for acute myocardial infarction (AMI). Ten of the 37 AMI patients (27%) underwent acute PTCA as primary treatment and seven patients (19%) received thrombolytic therapy for AMI despite prolonged (mean 24+/-13 min) cardiopulmonary resuscitation (CPR). The remaining 20 patients had no specific infarct treatment. Despite successful PTCA, in eight out of ten patients, their mortality in hospital was 60% (6/10). Mortality in the thrombolysis group was 57% (4/7). For the remaining 20 MI-patients the mortality was 65% (13/20). Univariate and multivariate analyses were performed to design a weighted prognostic scoring system. The Glasgow coma scale (GCS) was the strongest independent predictor (r=0.76, P< or =0.001) for in-hospital death. CONCLUSIONS: in-hospital mortality after successful OHR seems to largely depend on neurological status at admission and much less on the specific treatment of myocardial infarction. The prognostic scoring system accurately predicted the in-hospital mortality and can be used for early treatment stratification; however, it should be proven in a prospective study. PMID- 11008154 TI - Ventricular fibrillation exhibits dynamical properties and self-similarity. AB - Electrocardiographic recordings of ventricular fibrillation (VF) appear chaotic. Previous attempts to characterize the chaotic nature of VF have relied on peak-to peak intervals [Witkowski et al., Phys. Rev. Lett. 1995;75(6):1230-3; Garfinkel et al., J. Clin. Investig. 1997;99(2):305-314; Hastings et al., Proc. Natl. Acad. Sci. USA 1996;93:10495-9], the frequency spectrum [Goldberger et al., 1986;19:282 289] or other derived measures [Kaplan and Cohen, Circ. Res. 1990;67:886-92], with results that demonstrate some characteristics of chaos. We have sought to determine whether VF is chaotic rather than random and whether the waveform can be described quantitatively using the tools of fractal geometry. We have constructed an attractor, measured the correlation dimensions, estimated the embedding dimension and measured Lyapunov exponents. When the digitized waveform is analyzed directly, VF exhibits nonrandom, chaotic behavior over a decade of sampling frequency. Within the scaling range we have estimated the Hurst exponent, and the self-similarity dimension of the VF waveform, supporting the presence of chaotic dynamics. Furthermore, these characteristics are measurable in a porcine model of VF under different recording conditions, and in VF recordings taken from human subjects immediately prior to defibrillation. Analyses of the Hurst exponents and self-similarity dimensions are correlated with the duration of VF, which may have clinical applications. PMID- 11008155 TI - Ventilation volumes with different self-inflating bags with reference to the ERC guidelines for airway management: comparison of two compression techniques. AB - The 1998 ERC-guidelines for airway-management recommend an tidal volume of 400 600 ml for adults undergoing CPR. As commercially available self-inflating bags were designed to meet former recommendations (800-1200 ml) we investigated how to meet the latest recommendations with these bags. We combined the head of a training manikin (Laerdal Medical) and a standard lung (VTTL; Michigan Instrument), adjusted to a physiological compliance and resistance. Volume was measured with a Wright spirometer (BOC). Seven self-inflating bags were investigated. Tests were carried out by ten people (five female and five male) for 5 min each using two different techniques. Technique 1: standard ventilation with one hand without compression of the self-inflating bag against the rescuers knee. Technique 2: modified open palm technique with total squeezing of the self inflating bag by compression against the rescuers knee. The average tidal volumes for technique 1 ranged from 438 to 604 ml. Applying technique 2 the volumes ranged from 888 to 1192 ml. The latest recommendations were met using a single hand technique without compression against the rescuers knee for all seven bags tested. The modified open palm technique produced larger tidal volumes which were more in line with previous recommendations. PMID- 11008156 TI - Cardiopulmonary resuscitation skills in nurses and nursing students. AB - The purpose of this study is to describe the basic cardiac life-support (BLS) skills of nurses and nursing students in southern Finland and Hungary, and to assess the influence of resuscitation teaching and other group characteristics on performance. The data for the study were collected in the spring and autumn of 1997. The study group consisted of 75 nurses from Helsinki University Central Hospital's medical outcome unit, 188 final term students in four nursing institutes in Uusimaa county and 35 final term students in a Hungarian institute of nursing. A total of 298 people (34 men and 264 women) participated in the study. Background information was collected using a structured questionnaire devised specifically for this study. Resuscitation skills were measured using the Skillmeter Anne manikin. The manikin was placed supine during the test. After completing the questionnaire, every participant attempted resuscitation on the manikin - which was supplied with a printer - for 4 min. The results were printed out and attached to the questionnaires. The data were analysed using two-way frequency tables and logistic regression. Statistical differences were calculated using the chi(2)-test. The results showed that 53% of the participants had studied resuscitation during the last 6 months, but 7% had never participated in resuscitation teaching. Before testing, 55% of the participants estimated that their resuscitation skills were good. The results showed that 36% first assessed the patient's response, 67% opened the airway but only 3% determined pulselessness before starting to resuscitate. Twenty-one percent of the participants compressed correctly for at least half of the test and 33% ventilated correctly at least half of the time. Logistic regression showed that the best predictors for good response assessment skills went to those who were nursing students who had studied resuscitation skills sometime during the previous 6 months. The best predictor of the skill to open the airway was a positive attitude towards personal cardiopulmonary resuscitation (CPR) skills, i.e. self-confidence. The predictor for adequate skills in artificial ventilation was that they belonged in the group of nursing students who had benefited from recent resuscitation training (<6 months). In conclusion, the skills of the participants of the study can not be considered adequate in terms of an adequate and prompt assessment of the need for resuscitation, and a 50% success rate in artificial ventilation and chest compression. PMID- 11008157 TI - Effects of endothelin-1 on resuscitation rate during cardiac arrest. AB - OBJECTIVES: Endothelin-1 (ET-1) is a potent peripheral and coronary artery vasoconstrictor and has been shown to improve coronary perfusion pressure (CPP) during cardiac arrest. The effect of ET-1 on return of spontaneous circulation (ROSC) following cardiac arrest has not been studied. Our hypothesis was that ET 1 does not improve ROSC from cardiac arrest when compared to placebo. METHODS: A total of 11 immature swine were used in this laboratory study. Animals were randomized to receive 300 microg ET-1 and standard dose epinephrine (SDE) or placebo and SDE during arrest. After a 10-min period of no-flow ventricular fibrillation (VF), CPR was performed for 3 min followed by ET-1/SDE or placebo/SDE administration. Following drug administration, standard ACLS was followed with SDE given every 3 min. Aortic pressure was monitored during resuscitation. ROSC was defined as any perfusing rhythm with a systolic pressure greater than 60 mmHg for 60 s. Animals received post-ROSC care as needed for 2 h post-ROSC. CPP and ROSC were analyzed using repeated measures ANOVA and Fischer's exact test respectively. P<0.05 was considered significant. RESULTS: Pre-arrest variables and CPP prior to ET-1 administration were not different between groups. Following ET-1 administration, CPP was significantly increased at all time points in ET-1/SDE versus placebo/SDE animals. ROSC was achieved in 1/5 (20%) ET-1/SDE versus 1/6 (16.7%) placebo/SDE animals (P>0.05). The resuscitated ET-1/SDE animal survived 6.5 min compared to 120 min for the resuscitated placebo/SDE animal. CONCLUSIONS: In our study, ET-1 administration during cardiac arrest increases CPP but does not improve ROSC. PMID- 11008158 TI - Pentoxifylline improves circulatory and metabolic recovery after cardiopulmonary resuscitation. AB - BACKGROUND: To evaluate the effectiveness of a bolus application of pentoxifylline (PTXF) at the beginning of CPR in a standardized resuscitation animal model. METHODS AND RESULTS: In a laboratory model of cardiac arrest, 12 Wistar rats (382-413 g) were randomized into two groups. Both groups underwent 4 min of cardiopulmonary arrest induced by a transthoracic application of a fibrillating current of 10 mA. At the beginning of CPR, group one (n=6) received a bolus injection of 10 mg kg(-1) body weight PTXF versus sodium chloride in group two (controls: n=6). All animals developed a severe lactate acidosis during and after CPR but in PTXF treated animals acid-base values returned to baseline pattern. During return of spontaneous circulation (ROSC) in the PTXF group lactate concentration decreased from 13.4+/-2.1 to 1.9+/-0.7 mmol l(-1) within 60 min (P<0.01). In control animals, lactate values remained high (10.8+/-3.5 by 60 min, P<0.01). After bolus injection of PTXF pH increased from 6.93+/-0.06 to 7.29+/-0.13 within 60 min of ROSC versus 6.85+/-0.05 to 6.97+/-0.23 in sodium chloride treated animals (P<0.01). Within 5 min of ROSC, PTXF treated animals achieved higher oxygenation values (PTXF P(a)O(2)=216.9+/-62.5 mmHg, control 132. 2+/-15.1 mmHg, P<0.01). CONCLUSIONS: Administration of PTXF at the beginning of CPR improved macrocirculation, acid-base status and arterial oxygenation. PMID- 11008159 TI - Alteration of dopamine transport and dopamine D(2) receptor binding in the brain induced by early and late consequences of global ischaemia caused by cardiac arrest in the rat. AB - This study was designed to determine the effects of global cerebral ischaemia caused by temporary cardiac arrest and the early and late consequences of this ischaemia on dopamine transport and dopamine D(2) receptor binding in rat brain. The effects of 10 min of global ischaemia were measured immediately and after 1 h and 7 days post-resuscitation. A decrease of dopamine uptake in the rats by synaptosomes was noted immediately following global ischaemia and 1 h after resuscitation. However, at 7 days post-resuscitation, the dopamine uptake returned to control values. Reversibility of the changes in the synaptosomal dopamine uptake is undoubtedly a favourable sign. Global ischaemia and reperfusion after 1 h or 7 days did not show altered rates of dopamine release but did affect the dopamine D(2) receptor. An observed increase of receptor affinity may be an adaptive response to the reduction in binding capacity. The reduction of visible D(2) receptor binding sites in the early post-resuscitation phase, which was extended to the period of 7 days after resuscitation without recovery, is probably associated with neuronal necrotic damage. PMID- 11008160 TI - Catecholamines in cardiac arrest: role of alpha agonists, beta-adrenergic blockers and high-dose epinephrine. PMID- 11008161 TI - A persistent reduction in short-term facilitation accompanies long-term potentiation in the CA1 area in the intact hippocampus. AB - Exploration of the nature of the relationship between short-term and long-term synaptic plasticity should aid our understanding of their roles in brain function. The effects of inducing long-term potentiation on short-term facilitation at CA1 synapses in the stratum radiatum of the intact hippocampus were examined by recording the slope of the field excitatory postsynaptic potential in both urethane and freely behaving adult rats. Facilitation of the second synaptic response to paired-pulse stimulation (40ms interstimulus interval) was monitored before and after the induction of long-term potentiation by high-frequency stimulation (10 trains of 20 pulses at 200Hz). The tetanus triggered a rapid overall reduction in paired-pulse facilitation that persisted for at least 2h. In the anaesthetized animals a detailed correlation analysis revealed that initial paired-pulse facilitation level correlated strongly with the subsequent reduction in paired-pulse facilitation and the magnitude of long term potentiation. The reduction in paired-pulse facilitation also correlated with long-term potentiation magnitude. These relationships were not observed in animals with low initial degrees of paired-pulse facilitation. It was concluded that the relative contribution of different expression mechanisms of long-term potentiation varies depending on the initial facilitation characteristics of the synapses. Furthermore, the temporal selectivity and gain control of synapses can be altered persistently in the intact hippocampus. This suggests that there is considerable variation in the fidelity of temporal information storage at different synapses during learning and memory in the CA1 area. PMID- 11008162 TI - Re-structuring of synapses 24 hours after induction of long-term potentiation in the dentate gyrus of the rat hippocampus in vivo. AB - In male rats, long-term potentiation was induced unilaterally in the dentate gyrus, either by high frequency (200Hz) or theta rhythm stimulation. Structural synaptic changes were examined 24h after induction using quantitative electron microscopy. A disector technique was employed in order to estimate the density of synapses (using 70-80-nm sections) and of granule cell nuclei (using 2-microm sections) in the middle, and inner molecular layer in both hemispheres. Synaptic height and total lateral areas of synaptic active zones per unit tissue volume were assessed via assumption-free stereological techniques coupled with image analysis. The results obtained indicated that both synaptic density and number (corrected per neuron) of axo-spinous, but not axo-dendritic, synapses were approximately 40% higher in the middle, but not inner molecular layer of the potentiated hemisphere compared to the contralateral (control hemisphere). No significant inter-hemispheric difference was found in the volume densities of lateral areas of active zones. These data suggest that 24h after long-term potentiation induction, active zones of existing axo-spinous synapses either split forming separate contacts, or decrease in size while new synapses are formed. PMID- 11008163 TI - Temporal regularity of neurotransmitter release at single terminal in cultured hippocampal neurons. AB - The whole-cell GABA-mediated inhibitory postsynaptic currents were studied using the patch-clamp technique on synaptically connected cultured hippocampal neurons. The stimulus-evoked inhibitory postsynaptic currents were recorded in the tetrodotoxin-containing solution in response to the low-amplitude long (10-20ms) extracellular depolarization of a single presynaptic terminal. During each depolarization the postsynaptic response in a form of several superimposed independent events was recorded. The amplitudes of these responses fluctuated randomly, irrespective of the number of the event. In all the investigated neurons the distributions of delays revealed regularly spaced multiple peaks. The number of peaks increased with the duration of stimulus. The distance between the peaks was on average 2.97+/-0.86ms (n=58). The mean intervals between successive releases were distributed exponentially indicating the independence of the release sites. Thus neurotransmitter release might occur with maximal probability at the most probable times irrespective of the presence or failure of the previous event. The increase in stimulating pulse amplitude led to a decrease in the number of clearly detectable peaks in distributions. The decrease in the number of peaks in the distribution of delays was not accompanied with a decrease in the distance between peaks within the range of reliable resolution of the peaks. The amplitude distribution also revealed regularly spaced multiple peaks. The absence of significant correlation between the amplitude of the first and the second event demonstrated the independence of the succeeding release on the preceding release during long stimulation. Results of statistical analysis of our experimental data supports the hypothesis of multiquantal neurotransmitter release in a single inhibitory hippocampal synapse. Neurotransmitter release during long stimulation may occur at certain times with maximal probability, keeping the mean inter-release interval constant. Thus the interval is not determined directly by the depletion of vesicles and the number of vesicles which may be released at the most probable time is random. PMID- 11008164 TI - Accumulation of 4-hydroxynonenal-modified proteins in hippocampal CA1 pyramidal neurons precedes delayed neuronal damage in the gerbil brain. AB - It has been proposed that reactive oxygen species and lipid peroxidation have a role in the delayed neuronal death of pyramidal cells in the CA1 region. To explore the in situ localization and serial changes of 4-hydroxy-2-nonenal modified proteins, which are major products of membrane peroxidation, we used immunohistochemistry of the gerbil hippocampus after transient forebrain ischemia with or without preconditioning ischemia. The normal gerbil hippocampus showed weak immunoreactivity for 4-hydroxy-2-nonenal-modified proteins in the cytoplasm of CA1 pyramidal cells. 4-hydroxy-2-nonenal immunoreactivity showed no marked changes after preconditioning ischemia. In the early period after ischemia and reperfusion, there was a transient increase of nuclear 4-hydroxy-2-nonenal immunoreactivity in CA1 pyramidal neurons. In contrast, cytoplasmic immunoreactivity transiently disappeared during same period and then increased markedly from 8h to seven days. One week after ischemia, 4-hydroxy-2-nonenal immunoreactivity was observed within reactive astrocytes in the CA1 region. Early nuclear accumulation of 4-hydroxy-2-nonenal in CA1 neurons may indicate a possible role in signal transduction between the nucleus and cytoplasm/mitochondria, while delayed accumulation of 4-hydroxy-2-nonenal modified proteins in the cytoplasm may be related to mitochondrial damage. We conclude that 4-hydroxy-2-nonenal may be a key mediator of the oxidative stress induced neuronal signaling pathway and may have an important role in modifying delayed neuronal death. PMID- 11008165 TI - The hippocampus in spontaneously hypertensive rats: a quantitative microanatomical study. AB - The influence of hypertension on the morphology of hippocampus was assessed in spontaneously hypertensive rats of two, four and six months and in age-matched normotensive Wistar-Kyoto rats. Values of systolic pressure were slightly increased in two-month-old spontaneously hypertensive rats in comparison with age matched Wistar-Kyoto rats and augmented progressively with age in spontaneously hypertensive rats. No microanatomical changes were observed in the hippocampus of spontaneously hypertensive rats of two months in comparison with age-matched Wistar-Kyoto rats, whereas a decrease of white matter volume was observed in the CA(1) subfield and in the dentate gyrus of four-month-old spontaneously hypertensive rats. In the hippocampus of six-month-old spontaneously hypertensive rats a reduction of grey matter volume both in the CA(1) subfield and in the dentate gyrus, a loss of neurons affecting to a greater extent the CA(1) subfield and an increase of glial fibrillary acid protein-immunoreactive astrocytes was found. The occurrence of apoptosis and/or necrosis identified using the terminal deoxyribonucleotidyl transferase-mediated biotin-16-dUTP nick end labelling technique was also observed in the CA(1) subfield and to a lesser extent in the dentate gyrus. The only change noticeable in the CA(3) subfield of six-month-old spontaneously hypertensive rats was a slight increase in the number of glial fibrillary acid protein-immunoreactive astrocytes. These findings indicate the occurrence of neuronal loss and of astrocyte changes in the hippocampus of spontaneously hypertensive rats of six months, being the CA(1) subfield the area most affected. The relevance of these neurodegenerative changes in hypertension and the possible occurrence of apoptosis and/or necrosis as expression of hypertensive brain damage is discussed. PMID- 11008166 TI - Neuropeptides in hippocampus and cortex in transgenic mice overexpressing V717F beta-amyloid precursor protein--initial observations. AB - Immunohistochemistry was used to analyse 18- and 26-month-old transgenic mice overexpressing the human beta-amyloid precursor protein under the platelet derived growth factor-beta promoter with regard to presence and distribution of neuropeptides. In addition, antisera/antibodies to tyrosine hydroxylase, acetylcholinesterase, amyloid peptide, glial fibrillary acidic protein and microglial marker OX42 were used. These mice have been reported to exhibit extensive amyloid plaques in the hippocampus and cortex [Masliah et al. (1996) J. Neurosci. 16, 5795-5811]. The most pronounced changes were related to neuropeptides, whereas differences between wild-type and transgenic mice were less prominent with regard to tyrosine hydroxylase and acetylcholinesterase. The main findings were of two types; (i) involvement of peptide-containing neurites in amyloid beta-peptide positive plaques, and (ii) more generalized changes in peptide levels in specific layers, neuron populations and/or subregions in the hippocampal formation and ventral cortices. In contrast, the parietal and auditory cortices were comparatively less affected. The peptide immunoreactivities most strongly involved, both in plaques and in the generalized changes, were galanin, neuropeptide Y, cholecystokinin and enkephalin. This study shows that there is considerable variation both with regard to plaque load and peptide expression even among homozygotes of the same age. The most pronounced changes, predominantly increased peptide levels, were observed in two 26-month old homozygous mice, for example, galanin-, enkephalin- and cholecystokinin-like immunoreactivities in stratum lacunosum moleculare, and galanin, neuropeptide Y, enkephalin and dynorphin in mossy fibers. Many peptides also showed elevated levels in the ventral cortices. However, decreases were also observed. Thus, galanin-like immunoreactivity could not any longer be detected in the diffusely distributed (presumably noradrenergic) fiber network in all hippocampal and cortical layers, and dynorphin-like immunoreactivity was decreased in stratum moleculare, cholecystokinin-like immunoreactivity in mossy fibers and substance P like immunoreactivity in fibers around granule cells. The significance of generalized peptide changes is at present unclear. For example, the increase in the mainly inhibitory peptides galanin, neuropeptide Y, enkephalin and dynorphin and the decrease in the mainly excitatory peptide cholecystokinin in mossy fibers (and of substance P fibers around granule cells) indicate a shift in balance towards inhibition of the input to the CA3 pyramidal cell layer. Moreover, it may be speculated that the increase in levels of some of the peptides represents a reaction to nerve injury with the aim to counteract, in different ways, the consequences of injury, for example by exerting trophic actions. Further studies will be needed to establish to what extent these changes are typical for Alzheimer mouse models in general or are associated with the V717F mutation and/or the platelet-derived growth factor-beta promoter. PMID- 11008167 TI - Differential regulation of adult and embryonic glutamate decarboxylases in rat dentate granule cells after kainate-induced limbic seizures. AB - In adult brain, the inhibitory GABAergic neurons utilize two distinct molecular forms of the GABA-synthesizing enzyme glutamate decarboxylase (GAD), GAD65 and GAD67. During embryonic development, two truncated forms of GAD67 are also expressed (GAD25 and GAD44), which are translated from two embryonic-specific splice variants of GAD67 messenger RNA. It has recently been established that the excitatory dentate granule cells, in addition to the neurotransmitter glutamate, also contain low levels of GABA and GAD67, which are increased after limbic seizures. To study the seizure-induced activation of glutamate decarboxylase, we investigated the expression of both embryonic and adult glutamate decarboxylase messenger RNAs in the adult rat hippocampus after kainic acid administration by semi-quantitative reverse transcription-coupled polymerase chain reaction, in situ hybridization and immunoblotting. We observed a rapid induction of the embryonic glutamate decarboxylase messenger RNA in the granule cells of dentate gyrus. The expression of embryonic glutamate decarboxylase transcripts, identified here as the splice variant that contains exon 7/B, peaked at about 2h after kainic acid injection and gradually returned to nearly basal levels by 24h. Strikingly, this transient induction of embryonic glutamate decarboxylase messenger RNA was not accompanied by concomitant synthesis of its corresponding protein product GAD25. In contrast, the adult GAD67 messenger RNA and protein were both clearly up-regulated in granule cells, albeit with a certain delay, reaching a maximum around 4-6h after kainic acid injection and gradually returned to control levels by 24h. GAD65 remained unchanged at both messenger RNA and protein levels during the studied period. These characteristic and highly reproducible changes in the synthesis of glutamate decarboxylases indicate that GAD67 is the predominant form of glutamate decarboxylases involved in the elevated synthesis of GABA during seizures and suggest that the transient induction of the embryonic GAD67 messenger RNA that contains exon 7/B, but not GAD25 protein, may exert a role solely in the subsequent up-regulation of adult GAD67 transcription. Expression of the messenger RNA encoding for an alternatively spliced, truncated form of the GABA-synthesizing enzyme glutamate decarboxylase was detected in dentate granule cells briefly after kainic acid induced seizures. Just as during embryonic development, expression of the alternatively spliced messenger RNA was transient and followed by transcription of its adult form, indicating a possible recapitulation of an embryonic program of gene expression in adult granule cells after epileptic seizures. PMID- 11008168 TI - Expression of cholecystokinin, enkephalin, galanin and neuropeptide Y is markedly changed in the brain of the megencephaly mouse. AB - Megencephaly, enlarged brain, is a major sign in several human neurological diseases. The mouse model for megencephaly (mceph/mceph) has an enlarged brain, presumably due to brain cell hypertrophy, and exhibits neurological and motor disturbances with seizure-like activity, as well as disturbances in the insulin like growth factor system. Here, we report that expression of the neuropeptides cholecystokinin, enkephalin, galanin and neuropeptide Y is dramatically changed in mceph/mceph brains compared to wild type, as revealed by in situ hybridization and immunohistochemistry. The changes were confined to discrete brain regions and occurred in a parallel fashion for peptides and their transcripts. For cholecystokinin, mceph/mceph brains had region-specific up- and down-regulations in several layers of the hippocampal formation and increased levels in, especially ventral, cortical regions. Enkephalin messenger RNA expression was up regulated in the dentate gyrus granular layer and in ventral cortices, but down regulated in the CA1 pyramidal layer. Enkephalin-like immunoreactivity was elevated in mossy fibers of the hippocampus and the ventral cortices. Galanin expression was increased in several layers and interneurons of the hippocampal formation, as well as in ventral cortices. Galanin-like immunoreactivity was reduced in nerve terminals in the forebrain. Neuropeptide Y expression was increased in the hippocampal formation and ventral cortices. Whether the mainly increased peptide levels contribute to the excessive growth of the brain or represent a consequence of this growth and/or of the neurological and motor disturbances remains to be elucidated. PMID- 11008169 TI - Maternal corticosterone during lactation permanently affects brain corticosteroid receptors, stress response and behaviour in rat progeny. AB - The long-term consequences of a physiological-range increase of maternal corticosterone during lactation were investigated on the 15-month-old progeny. The offspring of rats drinking water supplemented with corticosterone (200 microgram/ml of corticosterone hemisuccinate) from day 1 postpartum to weaning exhibited: (i) better performance in a conditioned learning test; (ii) reduction of fearfulness in two conflict situations; (iii) lower stress-induced corticosterone secretion and (iv) higher number of corticosteroid receptors in the hippocampus. The results of this study show that the effects of maternal physiological-range hypercorticosteronemia during lactation are lifelong. Moreover, these data suggest that corticosteroids, secreted during neonatal life, may constitute a factor directing the neurobiological development of the infant. In line with this hypothesis, glucocorticoid-induced early events have consequences on the behavioral and physiological status of adulthood. These consequences may be either "beneficial" or "detrimental" depending on the plasma levels of corticosterone induced by the early life occurrences, as well as on the kind of the stimulus and the developmental stage at which the neonate experiences the event. The present study demonstrates that, when the increase of corticosterone in infancy is moderate, the adult rats show reduced anxiety, improved learning and a better coping strategy to deal with stressful situations. PMID- 11008170 TI - Reduced number of neurons in the hippocampus and the cerebellum in the postnatal guinea-pig following intrauterine growth-restriction. AB - Intrauterine growth restriction is a risk factor for neurological and behavioural deficits in children although the precise underlying biological correlate for this is unclear. The present study shows that animals with intrauterine growth restriction, induced by a period of reduced placental blood flow during the second half of pregnancy, demonstrate reduced numbers of neurons in the hippocampus and the cerebellum in conjunction with retarded dendritic and axonal growth within these structures. Intrauterine growth restriction was induced at 30 days gestational age by unilateral uterine artery ligation in pregnant guinea pigs. At one week of age, the total number of CA1 pyramidal neurons in the hippocampus and the Purkinje neurons in the cerebellum were determined using the combined fractionator/optical disector technique. The Cavalieri Principle was used to determine the volume of specific regions within the hippocampus and cerebellum. The body weight of animals that were classified as intrauterine growth-restricted was reduced by 42% (n=8) compared with control animals (n=8, P<0.001), while there was a smaller effect on brain weight (16% reduction, P<0.01). Estimates of the total number of neurons showed a reduction in CA1 pyramidal neurons in growth-restricted animals (4.19+/-0.43x10(5)) compared with control (5.20+/-0.44x10(5), P<0.01), and the volume of the stratum oriens layer above the CA1 region, which contains the apical dendrites of the CA1 pyramidal neurons, was reduced by 21% (P<0.01) in growth-restricted animals. In the cerebellum there was a reduction in the number of Purkinje neurons in growth restricted animals (3.97+/-0.50x10(5)) compared with control (5.13+/-0.52x10(5), P<0.01), and in the volume of the molecular layer (17%, P<0.05), the internal granular layer (22%, P<0.01) and in the volume of the cerebellar white matter (23%, P<0.01). These results show that a period of placental insufficiency during the second half of pregnancy can effect brain development in a way which could lead to neurological and behavioural deficits in the postnatal animal. PMID- 11008171 TI - Dopamine in the striatum modulates seizures in a genetic model of absence epilepsy in the rat. AB - Inhibition of the substantia nigra pars reticulata has been shown to suppress seizures in different animal models of epilepsy. The striatum is the main input of the substantia nigra pars reticulata. The aim of the present study was to examine the role of dopaminergic neurotransmission within the striatum in the control of absence seizures in a genetic model in the rat. Injections of mixed dopaminergic D1/D2 or of selective D1 or D2 agonists or antagonists in the dorsal parts of the striatum led to suppression of absence seizures associated with strong behavioral and electroencephalographic side-effects. When injected in the ventral part of the striatum (i.e. the nucleus accumbens core), all these agonists and antagonists respectively decreased and increased absence seizures without behavioral or electroencephalographic side-effects. Combined injections of low doses of a D1 and a D2 agonist in the core of the nucleus accumbens had an additive effect in absence seizures suppression. Furthermore, combined injections of low doses of a GABA(A) agonist and a N-methyl-D-aspartate antagonist in the substantia nigra also had cumulative effects in absence seizures suppression. These results show that dopamine neurotransmission in the core of the nucleus accumbens is critical in the control of absence seizures. The modulatory and additive effects on absence seizures of dopaminergic neurotransmission through both the D1 and D2 receptors in the core of the nucleus accumbens further suggest that ventral pathways of the basal ganglia system are involved in the modulation of absence seizures. PMID- 11008172 TI - Activated phosphorylation of cyclic AMP response element binding protein is associated with preservation of striatal neurons after focal cerebral ischemia in the rat. AB - Phosphorylation of the DNA-binding transcription factor, cyclic AMP response element binding protein, has recently been suggested to provide neuroprotective signals in times of cellular stress. Medium-sized striatal neurons are among the cells that are most vulnerable to ischemic stress in the brain. In the present study, phosphorylation of cyclic AMP response element binding protein was immunohistochemically evaluated in rat striatum in order to examine the ischemic vulnerability of each striatal region from the standpoint of cyclic AMP response element binding protein. Rats were subjected to 90-min focal cerebral ischemia followed by various periods of recirculation. Focal ischemia was induced by occlusion of the middle cerebral artery by the intraluminal suture method. Local cerebral blood flow measured by the 14C-iodoantipyrine method in the lateral and the medial striatal regions during occlusion was 5.0+/-7. 1 and 42.5+/ 8.1ml/100g/min, respectively. Cerebral blood flow in each region was restored to the control level during the recirculation period. The lateral and the medial regions of the striatum in the sham animals showed hardly any immunoreactivity with the specific antibody against phosphorylated cyclic AMP response element binding protein. By contrast, at 3.5h of recirculation, a number of phosphorylated cyclic AMP response element binding protein-positive neurons were detected in the medial striatal region on the occluded side, and the increase in the number of immunopositive cells continued until two weeks of recirculation with gradual decline. The lateral striatal region on the ischemic side showed only a mild increase in phosphorylated cyclic AMP response element binding protein-positive cells at 3.5h of recirculation, and the immunoreactivity rapidly disappeared during the subsequent recirculation period. Appreciable increase in immunoreactive cells was also noted in the contralateral striatum during the early phase of recirculation, and this increase seemed to be associated with spontaneous circling movements of the animals. Cresyl Violet staining revealed that striatal neurons in the medial region remained intact until two weeks of recirculation, whereas neurons in the lateral striatal region soon showed ischemic damage, followed by complete neuronal loss, and evolution of a frank infarct. Immunoreactivity for bcl-2, apoptosis-suppressive protein, was clearly detected in many neurons in the medial striatal region, but no such immunoreactivity was detected in the lateral striatal region. These findings suggest that persistently activated phosphorylation of cyclic AMP response element binding protein in the striatum during post-ischemic recirculation may be closely associated with protection of striatal neurons on the ischemic side, while it may be associated with spontaneous circling movements on the contralateral side. PMID- 11008173 TI - Facilitation of learning and long-term ventral pallidal-cortical cholinergic activation by proteoglycan biglycan and chondroitin sulfate C. AB - We have shown previously in the rat that biglycan, a recently discovered chondroitin sulfate proteoglycan, has neurotrophic effects which are mediated by its chondroitin/dermatan sulfate chains. Here we report that biglycan has neurochemical effects when injected into the nucleus basalis magnocellularis of the ventral pallidum, a site of dense cholinergic cell bodies. The effects on the cholinergic output in the frontal cortex are long lasting, indicating profound neuroactive function akin to that expected of a long-acting hormone. Injected into the same area of the brain, as well as into the ventricles in behaviorally impaired old animals, we found that biglycan can improve learning and memory in several behavioral paradigms. Furthermore, we show that both the neurochemical effectiveness as well as the promotion of learning is carried not by the proteoglycan per se, but rather by its chondroitin sulfate moiety, thus, demonstrating for the first time memory-promoting and neuroactive effects of a glycosaminoglycan, namely, chondroitin sulfate C. The results suggest that biglycan and other extracellular matrix molecules can have neurobehavioral and pharmacological functions for beyond those traditionally attributed to this class of molecules. PMID- 11008174 TI - Impaired spatial learning in aged rats is associated with loss of p75-positive neurons in the basal forebrain. AB - We investigated age-related changes in the number and size of neurons positive for the p75 neurotrophin receptor in the cholinergic basal forebrain of female Dark Agouti rats. Since the integrity of these neurons is known to be closely associated with performance in tests of spatial learning ability, we also investigated the incidence of age-related spatial learning impairments, using the Barnes maze. Spatial learning impairments occurred with increasing frequency with age. No rats showed impairment at six months, but 50% were impaired at 14 months and 71% at 26 months. There was no correlation between age and decreased number of p75-positive neurons in the rostral basal forebrain, which consists of the medial septum and vertical limb of the diagonal band of Broca. In the caudal basal forebrain, which consists of the horizontal limb and the nucleus of Meynert, there was a 13% reduction in the number of p75-positive neurons at 17 months compared to six months, and a 30% reduction at 26 months. There was a strong correlation between the presence of spatial learning impairment and a reduction in the number of p75-positive neurons. This correlation was most evident in the rostral basal forebrain, but was also present in the caudal basal forebrain. In the rostral basal forebrain, all learning impaired rats had fewer p75-positive neurons than the average number in unimpaired rats. A close correspondence between the presence of p75 and choline acetyltransferase was evident in basal forebrain neurons of learning-impaired and unimpaired rats. Gross pathological changes to the morphology of p75-positive neurons were relatively frequent in learning-impaired rats. These changes consisted of hypertrophy, appearance of vacuoles, and marginalisation of the cytoplasm. The results indicate the susceptibility of p75-positive neurons to degenerative changes with aging, and show that the loss of these neurons in the basal forebrain was strongly correlated with impairment in spatial learning. PMID- 11008175 TI - Contributions of mGlu1 and mGlu5 receptors to interactions with N-methyl-D aspartate receptor-mediated responses and nociceptive sensory responses of rat thalamic neurons. AB - The nociceptive responses of rat ventrobasal thalamus neurons can be reduced by N methyl-D-aspartate antagonists and by selective metabotropic glutamate receptor mGlu1 antagonists. The recent development of the mGlu5-selective antagonist 6 methyl-2-(phenylethynyl)-pyridine now allows the direct probing of the possible involvement of mGlu5 receptors in thalamic nociceptive responses. Extracellular recordings were made from single neurons in the ventrobasal thalamus and immediately overlying dorsal thalamic nuclei of adult urethane-anaesthetized rats using multi-barrel electrodes. Responses of neurons to iontophoretic applications of the mGlu5-selective agonist (R,S)-2-chloro-5-hydroxyphenylglycine were selectively reduced during continuous iontophoretic applications of 6-methyl-2 (phenylethynyl)-pyridine. Similar applications of 6-methyl-2-(phenylethynyl) pyridine reduced neuronal responses to noxious thermal stimuli to 53+/-9.5% of control responses. Co- application by iontophoresis of N-methyl-D-aspartate and metabotropic glutamate receptor agonists resulted in a mutual potentiation of excitatory responses. This effect could be reduced by either 6-methyl-2 (phenylethynyl)-pyridine or the mGlu1 antagonist LY367385. These results, taken together with previous data, suggest that acute thalamic nociceptive responses are mediated by a combination of mGlu1, mGlu5 and N-methyl-D-aspartate receptor activation, and that co-activation of these receptors produces a synergistic excitatory effect. Thus blockade of any of these receptor types would have a profound effect on the overall nociceptive response. PMID- 11008176 TI - Calcitonin gene-related peptide receptor expression in the neurons and glia of developing rat cerebellum: an autoradiographic and immunohistochemical analysis. AB - Quantitative autoradiography (using [125I]human alpha-calcitonin gene-related peptide as a ligand) and immunofluorescence (using monoclonal antibodies directed against a purified receptor) followed by confocal analysis were applied to analyse the distribution and cellular localization of the calcitonin gene-related peptide receptor in the rat cerebellum during development. From late embryonic days to the end of the second postnatal week, during the time window of calcitonin gene-related peptide expression in climbing fibers, high levels of calcitonin gene-related peptide binding sites were found in the white matter, where immunolabeling was present in oligodendrocytes. Lower levels were found in the cerebellar cortex, where receptor immunolabeling was found in Bergmann glia in a presumptive cell surface location and, during the second postnatal week, also in the cytoplasm of Purkinje cells. From the end of the second postnatal week to adulthood, when calcitonin gene-related peptide is no longer present in climbing fibers, the number of calcitonin gene-related peptide binding sites increased in the molecular layer, where not only Bergmann glia but also Purkinje cell distal dendritic branchlets were immunolabeled in a presumptive cell surface location. Concomitantly, the number of calcitonin gene-related peptide binding sites sharply decreased in the white matter. The developmental expression of the calcitonin gene-related peptide receptor and the previously described proliferating/differentiating effects of the peptide on glial cells suggest that calcitonin gene-related peptide and its receptor may promote a coordinated development of cerebellar glial cells, an effect driven mainly by the calcitonin gene-related peptide released by climbing fibers. As a result of glia-neuron interactions, an indirect effect on the differentiation of the cerebellar neuronal circuitry is also likely to occur. PMID- 11008177 TI - Group II and group III metabotropic glutamate receptor agonists depress synaptic transmission in the rat spinal cord dorsal horn. AB - The effects of group II and group III metabotropic glutamate receptor agonists on synaptic responses evoked by primary afferent stimulation in the dorsal horn, but mostly substantia gelatinosa, neurons were studied in the spinal cord slice preparation using conventional intracellular recording technique. Bath application of a potent metabotropic glutamate receptor 2- and 3-selective agonist (2S,1'R,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl) glycine reversibly suppressed monosynaptic and polysynaptic excitatory postsynaptic potentials evoked by A primary afferent fibers stimulation, the effect likely mediated by mGlu3 receptor subtype. This suppressing effect of (2S,1'R,2'R,3'R)-2-(2',3' dicarboxycyclopropyl) glycine on primary afferent neurotransmission was dose dependent and reduced by (S)-alpha-ethylglutamate, a group II metabotropic glutamate receptor antagonist. (2S,1'R,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl) glycine suppressed excitatory postsynaptic potentials without inducing detectable changes of postsynaptic membrane potential and neuronal input resistance in dorsal horn neurons. The paired-pulse depression at excitatory synapses between primary afferent fibers and dorsal horn neurons was reduced by (2S,1'R,2'R,3'R)-2 (2', 3'-dicarboxycyclopropyl) glycine application, suggesting a presynaptic site of action. The selective group III metabotropic glutamate receptor agonist (S)-2 amino-4-phosphonobutanoate also depressed A afferent fibers-evoked monosynaptic and polysynaptic excitatory postsynaptic potentials in a dose-dependent and reversible manner. The concentration-dependence of (S)-2-amino-4 phosphonobutanoate-mediated depression was most consistent with activation of mGlu receptor subtypes 4 and 7. However, on the basis of anatomical distribution of mGlu 4 and 7 subtypes, it is also possible that the (S)-2-amino-4 phosphonobatanoate effect is due to interaction with mGlu 7 receptor alone. (RS) alpha-cyclopropyl-4-phosphonophenylglycine a preferential antagonist at group III metabotropic glutamate receptors, completely reversed the depressant effects of (S)-2-amino-4-phosphonobutanoate on both monosynaptic and polysynaptic responses. (S)-2-amino-4-phosphonobutanoate reduced the paired-pulse depression at excitatory synapses between primary afferent fibers and dorsal horn neurons, but did not alter their postsynaptic membrane potential and input resistance. A clear facilitation of the (S)-2-amino-4-phosphonobutanoate-induced depression of monosynaptic and polysynaptic excitatory postsynaptic potentials in the absence of gamma-aminobutyric acid-subtype A receptor- and glycine-mediated synaptic inhibition was shown. Besides the depressant effect on excitatory synaptic transmission, inhibitory actions of group II and III metabotropic glutamate receptor agonists on the inhibitory postsynaptic potentials evoked by primary afferent stimulation in dorsal horn neurons were observed. These results suggest that group II and group III metabotropic glutamate receptors are expressed at primary afferent synapses in the dorsal horn region, and activation of the receptors suppresses synaptic transmission by an action on the presynaptic site. PMID- 11008178 TI - Mitogen activated protein kinase inhibition by PD98059 blocks nerve growth factor stimulated axonal outgrowth from adult mouse dorsal root ganglia in vitro. AB - Nerve growth factor stimulated axonal outgrowth from explanted mouse dorsal root ganglia is dependent on mitogen activated protein kinase. PD98059 ([2-(2'amino-3' methoxyphenyl)-oxanaphthalen-4-one]) blocks mitogen activated protein kinase by inhibiting its immediate upstream activator, mitogen activated protein kinase kinase (also known as MEK). Here we used PD98059 to study the role of mitogen activated protein kinase in the axonal outgrowth of adult dorsal root ganglia explants. Whereas PD98059 at 50 microM left spontaneous axonal outgrowth unaffected, it markedly inhibited nerve growth factor stimulated axon growth when assessed after two days in culture. A mitogen activated protein kinase assay and immunoblotting using antibodies discriminating between activated and inactivated kinase, both confirmed that PD98059 reduced the amount of activated enzyme in nerve growth factor stimulated preparations, while the total amounts of the kinase remained unchanged. Immunohistochemistry revealed the presence of neuronal mitogen activated protein kinase kinase and mitogen activated protein kinase itself. The latter enzyme was found to be activated in the growing axons, as seen by whole-mount labelling. At the ganglionic level activated mitogen activated protein kinase was preferentially detected in satellite cells. The results show that nerve growth factor stimulated axonal outgrowth in vitro from adult mouse dorsal root ganglia utilizes the mitogen activated protein kinase pathway. PMID- 11008179 TI - Nerve growth factor maintains potassium conductance after nerve injury in adult cutaneous afferent dorsal root ganglion neurons. AB - Whole-cell patch-clamp techniques were used to study the effects of nerve growth factor on voltage-dependent potassium conductance in normal and axotomized identified large cutaneous afferent dorsal root ganglion neurons (48-50 micrometer diameter) many of which probably give rise to myelinated Abeta fibers. K-currents were isolated by blocking Na- and Ca-currents with appropriate ion replacement and channel blockers. Separation of current components was achieved on the basis of response to variation in conditioning voltage. Cutaneous afferents were labeled by the retrograde marker hydroxy-stilbamide (FluoroGold) which was injected into the skin of the foot. The sciatic nerve was either ligated or crushed with fine forceps five to seven days later. Neurons were dissociated 14-17 days after injury. The cut ends of the sciatic nerves were positioned into polyethylene tubes, which were connected to mini-osmotic pumps filled with either nerve growth factor or sterile saline. Control neurons displayed a prominent sustained K-current and the transient potassium currents "A" and "D". Nerve ligation, which blocks target reconnection resulted in near 50% reduction of total outward current; isolated sustained K-current and transient A-current were reduced by a comparable amount. Nerve crush, which allows regeneration to peripheral targets and exposure of the regenerating nerve to the distal nerve segment, resulted in a small reduction in sustained K-current but no reduction in transient A-current compared to controls. Levels of transient A-current and sustained K-current were maintained at control levels after nerve growth factor treatment. These results indicate that the large reduction in transient A-current, and in sustained K-current, observed in cutaneous afferent cell bodies after nerve ligation is prevented by application of nerve growth factor. PMID- 11008180 TI - Fmr1 knockout mouse has a distinctive strain-specific learning impairment. AB - The Fmr1 gene knockout mouse is a model for the human Fragile X mental retardation syndrome. Fmr1 knockout mice with a C57BL/6-129/OlaHsd hybrid background have been reported to have only a very mild deficiency in learning the Morris water maze task. We compared the effect of this knockout mutation on learning in mice with either an FVB/N-129/OlaHsd hybrid background or a C57BL/6 background. When FVB-129 mice were tested in a cross-shaped water maze task, the knockout mice showed a pronounced deficiency in their ability to learn the position of a hidden escape platform in comparison to normal littermates. In contrast, knockout mice with a C57BL/6 background learned the maze just as well as their normal littermates. Fear conditioning did not reveal differences between knockout and normal mice in either background. These results show that silencing the Fmr1 gene clearly interfered with learning a specific visuospatial task in FVB/N-129 hybrid mice but not in C57BL/6 mice. The strain dependence may model the influence of genetic background in the human Fragile X syndrome. PMID- 11008181 TI - pH modulation of an inward rectifier chloride current in cultured rat cortical astrocytes. AB - The effects of changes in extra- and intracellular pH in the pathophysiological range (6.0-8.0) on astroglial plasma membrane ionic currents were investigated with the whole-cell patch-clamp technique. In cultured rat neocortical type-1 astrocytes differentiated by a long-term treatment with dibutyryl cyclic-AMP, exposure to an extracellular pH of 6.4 induced, as compared with the control extracellular pH at 7.3, a sustained and reversible increase in the holding current at -60mV. The rise in current was accompanied by a decrease in the apparent input resistance. Ion substitution experiments indicated that extracellular pH 6.4 upregulated the resting Cl(-) conductance, whereas an opposite effect could be observed at extracellular pH 8.0. Recordings of isolated Cl(-) currents showed that this modulation occurred on the previously identified hyperpolarization-activated, inwardly rectifying Cl(-) current, I(Clh). Extracellular acidification to pH 6.4 shifted the voltage dependence of I(Clh) activation by approximately 20mV towards more positive potentials, whereas a approximately 20mV opposite shift was observed upon exposure to extracellular pH 8.0. These effects were paralleled by an increase (extracellular pH 6.4) or decrease (extracellular pH 8.0) in the maximal conductance. Decreasing (6.0) or increasing (8.0) the intracellular pH shifted the steady-state activation of I(Clh) towards more negative or positive potentials, respectively, leaving unchanged the current sensitivity to extracellular pH modifications. The modulation of the inward rectifier Cl(-) current expressed by differentiated cultured neocortical astrocytes indicates that extra- and intracellular changes in pH occurring in a pathophysiological range may contribute to regulating Cl(-) accumulation in astroglial cells. PMID- 11008182 TI - Association of persistent falcine sinus with different clinicoradiologic conditions: MR imaging and MR angiography. AB - Nine pediatric patients are reported with persistent falcine sinuses associated with a variety of clinicoradiologic conditions and disorders. Besides MR imaging studies four patients had MR angiography examinations. Persistent falcine sinus was associated with arteriovenous malformations in three patients. In two cases persistent falcine sinus was an incidental finding. In the remaining four patients persistent falcine sinus was associated with total absence of the corpus callosum, acrocephalosyndactyly (Apert's syndrome), osteogenesis imperfecta, Chiari II malformation. The straight sinuses were either absent or rudimentary or entirely normal in these cases. It can be concluded that a mesenchymal disorder can be the primary cause for an open falcine sinus either in isolation or in association with variable changes in the straight sinus. PMID- 11008183 TI - MRI brain image segmentation by multi-resolution edge detection and region selection. AB - Combining both spatial and intensity information in image, we present an MRI brain image segmentation approach based on multi-resolution edge detection, region selection, and intensity threshold methods. The detection of white matter structure in brain is emphasized in this paper. First, a multi-resolution brain image representation and segmentation procedure based on a multi-scale image filtering method is presented. Given the nature of the structural connectivity and intensity homogeneity of brain tissues, region-based methods such as region growing and subtraction to segment the brain tissue structure from the multi resolution images are utilized. From the segmented structure, the region-of interest (ROI) image in the structure region is derived, and then a modified segmentation of the ROI based on an automatic threshold method using our threshold selection criterion is presented. Examples on both T1 and T2 weighted MRI brain image segmentation is presented, showing finer brain tissue structures. PMID- 11008184 TI - Wavelet based multiresolution expectation maximization image reconstruction algorithm for positron emission tomography. AB - Maximum Likelihood (ML) estimation based Expectation Maximization (EM) [IEEE Trans Med Imag, MI-1 (2) (1982) 113] reconstruction algorithm has shown to provide good quality reconstruction for positron emission tomography (PET). Our previous work [IEEE Trans Med Imag, 7(4) (1988) 273; Proc IEEE EMBS Conf, 20(2/6) (1998) 759] introduced the multigrid (MG) and multiresolution (MR) concept for PET image reconstruction using EM. This work transforms the MGEM and MREM algorithm to a Wavelet based Multiresolution EM (WMREM) algorithm by extending the concept of switching resolutions in both image and data spaces. The MR data space is generated by performing a 2D-wavelet transform on the acquired tube data that is used to reconstruct images at different spatial resolutions. Wavelet transform is used for MR reconstruction as well as adapted in the criterion for switching resolution levels. The advantage of the wavelet transform is that it provides very good frequency and spatial (time) localization and allows the use of these coarse resolution data spaces in the EM estimation process. The MR algorithm recovers low-frequency components of the reconstructed image at coarser resolutions in fewer iterations, reducing the number of iterations required at finer resolution to recover high-frequency components. This paper also presents the design of customized biorthogonal wavelet filters using the lifting method that are used for data decomposition and image reconstruction and compares them to other commonly known wavelets. PMID- 11008185 TI - Comparison of septal defects in 2D and 3D echocardiography using active contour models. AB - Three-dimensional ultrasound is emerging as a viable resource for the imaging of internal organs. Quantitative studies correlating ultrasonic volume measurements with MRI data continue to validate this modality as a more efficient alternative for 3D imaging studies. However, the processing required to form 3D images from a set of 2D images may result in a loss of spatial resolution and may give rise to artifacts. This paper examines a method of automatic feature extraction and data quantification in 3D data sets as compared with original 2D data. This work will implement an active contour algorithm to automatically extract the endocardial borders of septal defects in echocardiographic images, and compare the size of the defects in the original 2D images and the 3D data sets. PMID- 11008186 TI - Advances in three-dimensional reconstruction of the experimental spinal cord injury. AB - Three-dimensional (3D) computer reconstruction is an ideal tool for evaluating the centralized pathology of mammalian spinal cord injury (SCI) where multiple anatomical features are embedded within each other. Here, we evaluate three different reconstruction algorithms to three-dimensionally visualize SCIs. We also show for the first time, that determination of the volume and surface area of pathological features is possible using the reconstructed 3D images themselves. We compare these measurements to those calculated by older morphometric approaches. Finally, we demonstrate dynamic navigation into a 3D spinal cord reconstruction. PMID- 11008187 TI - A case of Streptococcus pneumoniae meningitis showing a marked contrast enhancement effect of whole arachnoid membrane on MRI. AB - We have experienced the adult case of meningitis caused by the Streptococcus pneumoniae, which showed the most aggressive clinical course. The peculiar and unique magnetic resonance images are reported. The whole arachnoid membrane was prominently enhanced following the contrast material injection, however, no contrast enhancement was observed in subarachnoid space, ependyma, intraventricular cerebrospinal fluid, and dura mater. This implies the superacute phase of the bacterial meningitis, in which inflammation is localized in the whole arachnoid membrane. The contrast enhancement effect in the meningitis may vary depending on the phase and severity, because it is a time-progressive disease. PMID- 11008188 TI - Semi-automated color segmentation of anatomical tissue, C. Imielinska, M. S. Downes, W. Yuan. Computerized medical imaging and graphics 2000;24(3):173-80 PMID- 11008189 TI - Preface to: Proceedings of the Fourth International Symposium on the Treatment of Parkinson's Disease. PMID- 11008190 TI - Parkinson's disease is not one disease. PMID- 11008191 TI - The role of D1-dopamine receptors in working memory-guided movements mediated by frontal cortical areas. AB - Like the striatum, the frontal motor cortices receive dopaminergic fibers from midbrain dopamine cells and contain high levels of dopamine receptors. Among frontal cortical areas, the dorsolateral PFC (PFd1) and the dorsal premotor cortex (PMd) have strong neural connections and play a major role for working memory-guided directional movements. To reveal the role of dopamine in this cognitive motor function, dopamine antagonists (SCH23390 for D1 receptors and sulpiride for D2 receptors) were applied locally or iontophoretically to the PFd1 and PMd in monkeys that performed delayed-response tasks with memory-guided directional movements. Applications of SCH23390, but not sulpiride, to these areas had significant effects at both the behavioral and neuronal levels. In the PFd1 and at the behavioral level, local injections of SCH23390 induced specific errors for memory-guided saccades, whereas it had no effects on visually guided saccades. In the PMd, local injections of SCH23390 induced directional errors and increased reaction time and movement time in memory-guided reaching movements. At the neuron level, iontophoretic applications of SCH23390 attenuated directional tuning of neurons of the PFd1 and PMd, which showed directional activities during the delay-and/or response-period(s). These findings suggest that the activation of D1-dopamine receptors in these frontal cortical areas plays a facilitating role in a series of neuronal processes of working memory-guided directional movements; the working memory process for guiding motor act in the PFd1 and preparation/control of directional manual movements in the PMd. In addition, our findings may provide insight into symptoms of schizophrenia and Parkinson's disease; the dysfunction of D1-dopamine receptors in the PMd1 and PMd may contribute to some symptoms, such as bradyphrenia and bradykinesia, in these disorders. PMID- 11008192 TI - Restless legs syndrome and its treatment by dopamine agonists. AB - The restless legs syndrome (RLS) characteristically presents with an irresistible urge to move that is most often accompanied by creeping sensations deep in the limbs. Occasionally the upper limbs can also be affected. RLS symptoms occur at rest and are typically more intense at night and at bedtime. Some patients complain about involuntary leg movements, so-called periodic limb movements (PLM), while at rest or PLM have been observed by the bed partner. Often, patients have to get out of bed several times at night, to relieve themselves of their disagreeable sensations.The prevalence of RLS is estimated to be about 5%. Up to now only three classes of drugs have been systematically evaluated for treatment of RLS: benzodiazepines, opioids and dopaminergic agents.The most consistent results have been obtained with dopaminergic drugs. Several studies have shown that L-dopa given with a peripheral decarboxylase inhibitor at a 10:4 ratio is effective in treating RLS. Controlled studies using polysomnographic recordings in a double-blind design showed that L-dopa administered at night produces a significant reduction of RLS occurring at bedtime and of PLM, which are often associated with nocturnal arousals. In most cases, L-dopa 100mg, in conjunction with the decarboxylase inhibitor carbidopa or benserazide 25mg, suppresses RLS although a rebound of PLM may be observed in the last part of the night. The two major adverse effects frequently seen in patients treated with L dopa are:Augmentation is one of the limiting factors of L-dopa therapy; thus, alternative treatment options are of major interest. In several open treatment trials performed with pergolide, patients reported a marked improvement of RLS symptoms including sleep problems. Mild symptoms of augmentation under pergolide treatment have been reported from single patients. In another 6-month open label trial, pergolide proved to be effective in patients who developed augmentation under L-dopa by relieving daytime symptoms after switching to pergolide.Most recently, the results of these open label trials have been replicated in a randomized, placebo-controlled, double-blind multicenter trial. Treatment with a single evening dose of 0.25-0.75mg pergolide resulted in a significant improvement of almost all subjective and objective parameters. Under pergolide, patients rated their RLS symptoms and sleep disturbances much less severe and polysomnographic recordings also revealed a significant improvement of all important sleep parameters. To prevent peripheral side-effects such as nausea or orthostatic hypotension, pergolide should be slowly up-titrated or domperidone should be added. Under these conditions, no major side-effects have been observed in treatment trials with pergolide in dosages up to 1.25mg.Pergolide with a half life of 12-16h thus appears to be an appropriate drug in the therapy of RLS especially in those patients who developed augmentation under L-dopa therapy. Owing to the remarkable therapeutic effect of pergolide on RLS symptom control, other dopamine agonists are presently being tested for the treatment of RLS. PMID- 11008193 TI - The early treatment of Parkinson's disease: levodopa, dopamine agonists or both. AB - Much has been written about the pharmacologic management of Parkinson's disease (PD) because of an expanding arsenal of antiparkinson drugs and our quest to alter the natural history of disease. Choice of initial therapy may prove fundamental to a treatment strategy that maximizes symptomatic control while minimizing the chances for long-term complications such as motor fluctuations and dyskinesias. Dopamine agonists (DA) have assumed a primary role in the early therapy of PD because of their antiparkinson effectiveness and low propensity to induce fluctuations and dyskinesias. Four available DA in the United States and an array of recent studies supporting their utility in early PD have shaped current PD management. Moreover, DA have neuroprotective properties in vivo. Nevertheless, levodopa remains the most effective drug for symptomatic control in PD. There is conflicting evidence regarding the putative neurotoxicity of levodopa and the mechanisms for levodopa-related motor fluctuations are not entirely known. Clinicians must therefore weigh the available evidence as they initiate therapy in PD. Clearly, both DA and levodopa will remain essential components of the early management of PD. PMID- 11008194 TI - Double blinded evaluation of the effects of pallidal and subthalamic nucleus stimulation on daytime activity in advanced Parkinson's disease. AB - The results of a double blinded evaluation of the effects of globus pallidus (GPi; n=7) and subthalamic nucleus (STN; n=11) stimulation in patients with advanced Parkinson's disease are summarized. The patients were evaluated at 6 8months after surgery. In order to determine the benefits afforded by the stimulation to the actual daily activities, the patients were maintained on medication with optimal doses and schedules. The stimulation was turned off overnight for at least 12h. It was turned on in the morning (or maintained turned off), and the best and worst scores during daytime activity were recorded, as on period and off-period scores, respectively. A reduction in total motor score on the Unified Parkinson's Disease Rating Scale was clearly elicited by GPi and STN stimulation at both the off-period (-57 and -29%, respectively) and the on-period (-36 and -25%, respectively). The difference in effects between GPi and STN stimulation appeared to be due largely to an unintended difference in the patients' preoperative symptoms. The benefits provided by stimulation to the actual daily activities appears to be limited in patients who have become unresponsive to a large dose of levodopa. Two advantages of GPi and STN stimulation were identified. Firstly, the stimulation can supplement a reduced action of levodopa during the off-period. It thus improves the patient's daily activities through attenuation of the motor fluctuations. Secondly, the stimulation can replace part of the action of levodopa during the on-period. It thus attenuates dopa-induced dyskinesia through a reduced dose of medication. More importantly, the stimulation improves the daily activities in dopa intolerant patients who are being administered a small dose of levodopa because of unbearable side effects. In addition, GPi stimulation has its own inhibitory effect on dopa-induced dyskinesia. Clinically important improvement was observed in severe gait freezing in 2 patients following unilateral anterodorsal GPi stimulation on the right side alone. PMID- 11008195 TI - Management of behavioral and psychiatric problems in Parkinson's disease. AB - Behavioral and psychiatric problems associated with idiopathic Parkinson's disease (PD) include cognitive dysfunction, drug-related psychosis, depression, anxiety, apathy, fatigue and sleep disturbance. These nonmotor symptoms are a significant cause of disability at all stages of illness. Cognitive dysfunction spans a continuum from circumscribed cognitive impairments to severe global dementia which can occur in up to 10-30% of advanced PD patients. Psychosis develops in 20-30% of PD patients receiving chronic antiparkinsonian therapy. Visual hallucinations and paranoid delusions are the most frequent symptoms. The gradual elimination of drugs of lesser priority that may affect cognition and/or cloud the sensorium constitutes the first step in the management of cognitive and psychotic symptoms. Atypical neuroleptic agents are an invaluable tool in those cases in which maximum drug regimen simplification is not adequate or results in unacceptable immobility. Depression and anxiety often go unrecognized although they are eminently treatable and may be important contributors to the morbidity of PD. They are present in 30-40% of PD patients and frequently occur together in association with other nonmotor symptoms such as apathy, fatigue and sleep disturbance. A combination of early recognition, counseling, antidepressant therapy, antianxiety and well-balanced antiparkinsonian therapy sets the stage for improved quality of life for patients with PD. PMID- 11008196 TI - Dopamine agonists: the treatment for Parkinson's disease in the XXI century? AB - Levodopa combined with a peripheral dopa-decarboxylase inhibitor (DCI) has been considered the therapy of choice for Parkinson's disease (PD). Levodopa is nearly always effective, but has a high incidence of adverse effects with long term use, including response fluctuations (on/off phenomena) and dyskinesias. Dopaminergic agonists, acting directly at the receptor level, would be able to decrease the incidence of these motor complications.In progressive neurodegenerative diseases, such as PD, modification of the rate of disease progression (often referred to as neuroprotection) is currently a highly debated topic. Increased oxidative stress is thought to be involved in nigral cell death, that is characteristic of PD. This oxidative stress may be further exacerbated by levodopa therapy. These mechanisms have been proven in vitro and animal models, but it's relevance in humans remains speculative.Based on the considerations above, the emerging therapeutic strategies for PD advocate early use of dopamine agonists in the treatment of PD. A number of recent well-controlled studies have proven the efficacy of dopamine agonists used as monotherapy. Moreover, as predicted by animal studies, on the long term, dopaminergic agonists induce significantly less motor complications than levodopa.In the last 2years, three new dopamine agonists have been launched, including ropinirole, pramipexole and cabergoline. These new agonists have been added, as therapeutical options to well-established drugs, like pergolide, bromocriptine or talipexole. The recently launched compounds have proven efficacy in monotherapy and as adjunctive therapy to levodopa. Unfortunately, only a very limited amount of comparative data among the different agonists is available. Pergolide has proven to be a superior drug to bromocriptine as adjunctive therapy to levodopa in a significant number of studies and is considered the gold standard dopamine agonist. Nevertheless, none of the recently launched compounds has compared itself against pergolide.A comparison of monotherapy trials is difficult, because of differences in design and populations. In a recently completed trial pergolide was statistically significantly better than placebo in all the efficacy parameters tested, with 57% of pergolide treated patients improving over 30% in the motor section of the UPDRS, as compared to 17% in the placebo arm. Interestingly, these results were obtained in the absence of any other antiparkinsonian drug during the trial. Recent monotherapy trials done with ropinirole and pramipexole achieved also significant improvements as monotherapy, but in these cases selegeline, a drug that causes a symptomatic improvement in PD, was allowed as co-medications during the trial. Not all trials used the same efficacy measures, i.e. monotherapy trials with pergolide and ropinirole used a "responder" based analysis (responder were all patients that improved 30% or more on the motor section of UPDRS), as well as a baseline to endpoint improvement in motor scores. Pramipexole monotherapy trials used only the latter approach, which is clinically less powerful than a responder analysis.Even with the difficulties mentioned above, all the recent trials with dopamine agonists have proven that these drugs are a useful symptomatic long term treatment for PD with or without levodopa and that the early use of dopamine agonists reduces the incidence of motor complications as compared to levodopa. PMID- 11008197 TI - Treatment of early Parkinson's disease. AB - Levodopa is still the most effective therapeutic agent for the treatment of Parkinson's disease (PD). Initially, levodopa provides a stable therapeutic response but, during long-term treatment its beneficial effect declines and a gradually increasing number of patients experience fluctuations in motor response. Therefore, in the management of PD it is important to minimise the risks for the development of motor fluctuations. In this context, recent double blind long-term studies have confirmed the earlier results, suggesting that it appears advisable to initiate dopaminergic treatment in early PD by initially using a dopamine agonist and by adding levodopa when the benefit is no longer adequate with dopamine agonist alone. Another alternative would be to start with selegiline alone, then depending on the disability of the patient, add a dopamine agonist and finally levodopa. PMID- 11008198 TI - Diagnosis and differential diagnosis of Parkinson's disease and parkinsonism. AB - In the clinic setting, most cases represent either Parkinson's disease (PD) or one of the other neurodegenerative disorders that make up the parkinsonism-plus syndromes. The major parkinsonism-plus syndromes include progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, as well as parkinsonism occurring in the context of Alzheimer's disease or one of the other primary dementing disorders. There are a variety of other conditions, however, that occasionally come into the differential diagnosis. These fall into the categories of secondary parkinsonism (due to drugs, toxins, structural lesions, etc.), another tremor syndrome such as essential tremor, or a hereditary disorder with parkinsonism. This broad differential diagnosis is reviewed. PMID- 11008199 TI - Treatment of Parkinson's disease in Japan. AB - In this paper, the course of therapy for Parkinson's disease is outlined. The rationale for the use of DA-receptor-agonist (DA agonist) monotherapy or early combination therapy using levodopa and a DA agonist is that these therapies are asociated with a lower incidence of motor complications. However, the disease progresses, the use of levodopa in combination with a DA agonist results in motor complications and development of levodopa dependency in parkinsonian patients, because the effect of levodopa on parkinsonism is very strong. In this study, a positive correlation between the Hoehn-Yahr severity score at off-periods and duration of illness was observed in parkinsonian patients with long duration of illness. This indicates that responsiveness to dopaminergic therapy still exists even in patients in advanced stages of Parkinson's disease, indicating that continuous stimulation of DA receptors and reducing the excessive fluctuation in the plasma levodopa level possibly improve motor complications. If the dose of the DA agonist is simply increased without reduction of levodopa doses, dyskinesia worsens. Although levodopa therapy is essential in the case of patients in advanced stages of Parkinson's disease, the therapeutic principle, which depends on levodopa efficacy, must be changed. Reduction of the levodopa dose and administration of a sufficient dose of a DA agonist, which is equivalent to levodopa dose reduction, is one of the possible means of effective therapy of the disease. PMID- 11008200 TI - C-MYC expression in medulloblastoma and its prognostic value. AB - To identify prognostic factors in medulloblastoma, a common malignant brain tumor of childhood, expression of the oncogene c-myc was examined at the mRNA level by in situ hybridization. c-myc mRNA expression was observed in 30 of 72 tumors (42%). The c-myc gene copy number was determined by quantitative PCR from genomic DNA of paraffin-embedded tumors. c-myc gene amplification was present in 5 of 62 cases (8.3%). Therefore, c-myc amplification was obviously not the cause of c-myc mRNA expression in most samples. Kaplan-Meier estimation revealed a significant correlation between c-myc mRNA expression and survival (total mean follow-up 4.6 +/- 3.6 years, log-rank p = 0.02). Multivariate logistic regression analysis including sex, age, histological type, degree of surgical resection and expression of synaptophysin, GFAP and c-myc, was carried out on 54 patients who received both radiotherapy and chemotherapy. The analysis identified expression of c-myc as an independent predictive factor of death from disease. PMID- 11008201 TI - Gene amplifications detected by fluorescence in situ hybridization in pure intraductal breast carcinomas: relation to morphology, cell proliferation and expression of breast cancer-related genes. AB - Investigation of early breast carcinogenesis is limited by the difficulty in obtaining cell cultures or adequate fresh frozen material and by the fact that available data from in situ techniques are interpreted in terms of various classification systems. Our studies in a series of pure ductal carcinomas in situ (DCIS) were conducted in accordance with the recommendations of the international Consensus Conference (Hum. Pathol., 28, 122-125, 1997) relative to processing, determination of lesion extent, and histological stratification primarily on nuclear grade (NG). A multifactorial study performed in 15 low- and 16 high-NG DCIS (68% detected by mammography) included the following: (1) morphological analysis of NG, necrosis, and architectural pattern; (2) detection of numerical genomic abnormalities at ERBB2, MYC, CCND1, Xq1.2 and 20q13 loci by fluorescence in situ hybridization on interphase nuclei; and (3) immunohistochemical determination of cell proliferation, p53 accumulation, hormonal receptors and bcl 2 expression on serial sections of formalin-fixed, paraffin-embedded specimens. High NG, comedo/solid pattern and necrosis were significantly associated with amplification at one or more loci, the number of amplified loci, amplification at the ERBB2 locus, absence of bcl-2 and hormonal receptor expression and high cell proliferation (p < 0.05). High NG and comedo/solid pattern were significantly associated with MYC amplification and p53 accumulation, and necrosis with CCND1 amplification (the only gene amplification detected in low NG DCIS). These data provide additional information on the early steps of breast carcinogenesis, in accordance with currently recognized criteria of histological classification. PMID- 11008202 TI - Rapid quantitative detection of carcinoembryonic antigen-expressing free tumor cells in the peritoneal cavity of gastric-cancer patients with real-time RT-PCR on the lightcycler. AB - Detection of free cancer cells in the peritoneal cavity by RT-PCR using carcinoembryonic antigen (CEA) as a target gene is a more sensitive predictor of peritoneal dissemination than conventional cytology in gastric-cancer patients. Difficulties with this method are the lack of quantitative assessment of free cancer cells and the length of time before completion. To overcome these problems, we have established a rapid and quantitative detection method using a novel real-time fluorescence PCR system (LightCycler). Using this device with hybridization probes as fluorophores, we detected CEA mRNA in peritoneal washes during surgery (within 3 hr) without any post-PCR procedure. This method could reproducibly quantitate 10 to 10(6) CEA-expressing colon carcinoma cells per 10(7) peripheral blood leukocytes, a comparable sensitivity to conventional RT PCR with a wide dynamic range. Analysis of peritoneal washes from 109 gastric cancer patients with this assay revealed relative values of CEA transcripts that correlated well with the depth of tumor invasion (p < 0.01). Average values of CEA transcript in peritoneal washes in patients with cytology (-)/RT-PCR(-), cytology (-)/RT-PCR(+) and cytology (+)/RT-PCR(+) results were 0.64, 1,525 and 6,715, respectively. Moreover, CEA transcripts in peritoneal washes in patients with synchronous peritoneal metastasis were more than 50-fold higher than in those without metastasis. These results suggest a positive correlation between CEA mRNA levels in peritoneal washes and prognosis. We conclude that real-time RT PCR with hybridization probes is a sensitive, quantitative, specific and rapid method to detect free cancer cells in peritoneal washes. This clinically relevant system is a powerful technique to evaluate the risk of peritoneal recurrence in patients with gastric cancer. PMID- 11008203 TI - Expression of glypican 3 (GPC3) in embryonal tumors. AB - Embryonal tumors, such as neuroblastoma, medulloblastoma and Wilms' tumor, have their peak incidence in the first 4 years of life. These neoplasias exhibit genetic and clinical heterogeneity, but little is known about their molecular pathogenesis. Application of the differential-display PCR approach led to the identification of a gene, glypican 3 (GPC3), that is differentially expressed in cancer cells. Expression of this gene is usually limited to fetal mesodermal tissue, and its inactivation has been found to be responsible for the X-linked Simpson-Golabi-Behmel overgrowth syndrome. Here, we show that GPC3 mRNA is present in several neuroblastomas and all Wilms' tumors tested to date but not in medulloblastoma. GPC3 was not expressed in normal kidney tissues obtained from the corresponding Wilms' tumor patients, suggesting that in these cancer cells expression was not repressed (or was activated). No correlation was found between expression of GPC3 and the known indicator of neuroblastoma prognosis MYCN mRNA. However, all samples that expressed GPC3 also expressed IGF-II, coding for a growth factor important in the survival and growth of many cancer types. Although the biological significance of this relationship remains unclear, our results suggest that GPC3 may be implicated in the development of embryonal tumors through a signaling pathway that appears to involve IGF-II. PMID- 11008204 TI - Bcl-2 expression is associated with improved prognosis in patients with distal colorectal adenocarcinomas. AB - Several recent studies have suggested that the anatomic location of the tumor and ethnicity of the patient should be considered in the evaluation of prognostic markers of colorectal neoplasia. The phenotypic expression of Bcl-2 has been reported to be a useful prognostic marker in colorectal adenocarcinoma (CRC). However, its prognostic importance in CRCs based on their anatomic location and on the ethnicity of the patients has not been reported. Therefore, we evaluated Bcl-2 expression by immunohistochemistry in CRCs collected from 107 African American and 149 Caucasian patients from a southern U.S. population. In univariate Kaplan-Meier survival analyses, Bcl-2 expression was associated with better overall survival of both African-American (log-rank, p = 0.040) and Caucasian (log-rank, p = 0.032) patients with distal but not with proximal CRCs. In multivariate Cox regression analyses, when the pathologic features of tumors were not included, the expression of Bcl-2 was associated with better survival in either ethnic patient populations with distal CRCs after adjusting for other confounding variables and p53(nac) status; however, it was not significant in either race when tumor stage was included in multivariate analyses. Thus, these studies suggest that the expression of Bcl-2 in CRCs is a valuable indicator of good prognosis in either race when CRCs are located in the distal colorectum. Also, these studies suggest that the expression of Bcl-2 is useful in determining prognosis before pathologic-clinical staging and it can aid in selection of treatment after evaluation of diagnostic biopsy specimens of patients with distal colorectal adenocarcinomas. PMID- 11008205 TI - Urokinase-type plasminogen activator and its receptor in colorectal cancer: independent prognostic factors of metastasis and cancer-specific survival and potential therapeutic targets. AB - Urokinase-type plasminogen activator (uPA) and its receptor (uPAR), plasminogen (Plg), and plasminogen activator inhibitors-1 and -2 (PAI-1 and PAI-2) have been observed in many cancers and may contribute to progression and metastasis. In our study, we examined the expression of the 5 proteins by immunohistochemistry in 59 consecutive primary colorectal cancers (CRC) and correlated the protein expression with patient outcome. In addition, we determined the effect of down regulation of uPAR on the invasive/metastatic capability of CRC cells, by measuring antisense-uPAR transfected HCT116 and control cell lines, in terms of uPAR expression, uPA-binding activity, invasiveness through Matrigel in vitro and metastasis after cecal orthotopic implantation in nude mice in vivo. We found that higher expression of uPA or uPAR in primary tumor tissues was positively correlated with distant metastasis of CRC (Mann-Whitney, p < 0.02) and negatively correlated with both patient overall survival (OS) and cancer-specific survival (CSS; Cox model, p < 0.04). The prognostic value of uPA and uPAR for both OS and CSS was independent of other variables (multivariate Cox model, p < 0. 007). Antisense-uPAR transfected HCT116 cells, which expressed significantly lower levels of total cellular and cell surface uPAR proteins and uPA-binding activity compared with either wild-type or cells transfected with vector alone (Bonferroni, p < 0.05/3), consistently showed decreased invasiveness through Matrigel (Bonferroni, p < 0.05/3) and decreased metastasis formation in nude mice (Fisher, p < 0.05). Our data suggest that uPAR and uPA are independent prognostic factors in CRC; anti-uPAR treatment, which affects both uPAR and uPA levels, may have potential for new treatment of the disease. PMID- 11008206 TI - Nucleolus organizer regions (AgNORs) and total tumor mass are independent prognostic parameters for treatment-free period in chronic lymphocytic leukemia. AB - The aim of our study was to evaluate whether the nucleolus organizer regions (AgNOR) of circulating lymphocytes in chronic lymphocytic leukemia (CLL) could predict the duration of the stable phase (treatment-free period) of this disease. Age, sex, peripheral lymphoctes, hemoglobin, platelet count, Binet's stage, total tumor mass (TTM) and AgNOR pattern at diagnosis were assessed in 57 patients and compared with the time from diagnosis until patients fulfilled the criteria to start chemotherapy. In univariate Cox regression, Binet's stage, hemoglobin, number of peripheral lymphocytes, TTM and the percentage of lymphocytes containing one AgNOR cluster (circulating proliferative fraction) had predictive value. In the multivariate Cox model, only TTM and the percentage of cells with one AgNOR cluster were independent factors predicting the duration of the stable phase. Repetition of these analyses in 500 data sets created by bootstrap resampling confirmed the results. Summing up the percentage of lymphocytes with an AgNOR cluster and the TTM value, a simple prognostic index could be created to give information about the duration of the stable phase in CLL. PMID- 11008207 TI - Microsatellite instability, prognosis and metastasis in gastric cancers from a low-risk population. AB - We examined 169 cases of gastric adenocarcinoma for microsatellite instability (MSI), using a panel of 8 microsatellite markers. Of these cases, 142 were from the United States, a country of relatively low risk for gastric cancer. Comparing microdissected tumors to normal cells from the same patient, we classified tumors as being microsatellite-stable (MSS) or having a low frequency of MSI (MSI-L, up to 30% of markers different in the tumor) or a high frequency of MSI (MSI-H, 30% or more of markers different). Among our American cases, we identified 26 (18.2%) showing MSI-H and 15 (10.6%) showing MSI-L. Twenty cases were from Korean patients, and they showed no significant differences in proportions of MSI-H and MSI-L from the American cases. MSI-H tumors in the American patients were characterized by elevated frequencies of band shifts in repeat sequences of the BAX (50%), transforming growth factor-beta receptor type II (TGFbetaRII, 68.9%), beta(2)-microglobulin (21.4%) and E2F4 (51.7%) genes. Alterations in E2F4 in MSI H tumors were always integral multiples of 3 nucleotides lost or gained, which would not cause a frameshift mutation, and within the range of normal polymorphisms for this sequence. North American patients (n = 127) with MSI-H and MSI-L tumors had a longer median survival of 541 days and 587 days, respectively, compared to 265 days for patients with MSS tumors (p = 0.027). This survival difference may result from a significantly greater tendency for metastases in the MSS group (p = 0.031). PMID- 11008208 TI - ERCC1 expression as a molecular marker of cisplatin resistance in human cervical tumor cells. AB - Cisplatin is a valuable adjuvant to radiotherapy for the treatment of cervical cancer. Because the advantage of combining cisplatin with radiotherapy is likely to be attributable to additive cell killing by these 2 agents, such protocols should primarily benefit patients who have inherently cisplatin-sensitive tumors. Development of a molecular assay to rapidly evaluate the cisplatin responsiveness of cervical tumors would thus be extremely valuable. We investigated whether high pre-treatment mRNA levels of the ERCC1 nucleotide excision repair gene are predictive of cisplatin resistance in early-passage human cervical cancer cells, as they are in several other tumor types. Expression of the ERCC1 gene at the mRNA and protein levels was established by Northern and Western blotting, respectively, in a panel of single-cell-derived cervical carcinoma cell lines that exhibited a wide range of inherent sensitivity to cisplatin. There was a significant (p 12 h) of hypoxia. Astrocytes did not produce any significant amount of MIP-2 even though astrocytes maintained 98-99% viability following H12/R24. We also found that microglia survived the H/R treatment better (following H24) in the presence of astrocytes (mixed glial culture) than in microglia-enriched culture. Reoxygenation for prolonged periods (3 and 5 days) following H24 resulted in progressively larger increases in MIP-2 production (20- and 60-fold, respectively) in mixed glial cultures. Immunocytochemical staining revealed that the cells expressing MIP-2 in response to H/R were microglia rather than astrocytes in mixed glial cultures. Examination of MIP-2 mRNA expression showed that H/R upregulated MIP-2 gene expression. Taken together, our data suggest that microglial cells are an important source of MIP-2 production and suggest a potential injury mechanism involving brain-derived production of MIP-2 in H/R. PMID- 11008216 TI - Gene expression of the transporters and biosynthetic enzymes of the osmolytes in astrocyte primary cultures exposed to hyperosmotic conditions. AB - Sorbitol, myo-inositol, betaine, and taurine are held as organic osmolytes. When cells are exposed to a hyperosmotic medium, they accumulate these organic compounds and thus achieve osmotic equilibrium with the medium while maintaining their volume. In astrocyte primary cultures adapted to a chemically defined medium and then exposed to a medium made 30% hyperosmotic by adding sodium chloride or raffinose, we have comparatively investigated the expression of the genes encoding the proteins that control the cellular accumulation of these osmolytes, namely sorbitol biosynthetic enzyme, aldose reductase (AR), taurine biosynthetic enzymes, cysteine dioxygenase (CDO), and cysteine sulfinic acid decarboxylase (CSD), and the transporters of taurine (TauT), myo-inositol (SMIT), and betaine (BGT1) by assaying the corresponding mRNA levels through relative quantitative RT-PCR. When exposed to the hyperosmotic medium the astrocytes shrank rapidly and then slowly regained their initial volume after several hours. CDO- and CSD-mRNA remained unchanged, whereas AR-mRNA appeared increased only with the medium made hyperosmotic with sodium chloride. The mRNA levels of the transporters only showed significant and comparable increases in both hyperosmotic conditions. They were all significantly higher after 4-h exposure and back or close to normal values after 24-h exposure. The maximum level occurred at around 4 h (SMIT), 8 h (BGT1), and 12 h (TauT). The amplitude of BGT1 mRNA increase was much larger. When taurine was added to the hyperosmotic medium the cell volume recovery was greatly accelerated and the osmo-induced overexpression of TauT-, SMIT-, and BGT1-mRNA was fully prevented. The activation of the genes encoding the osmolyte transporters appears to be triggered when the cell shrinks below a certain volume threshold and prolonged once the cell volume has regained this threshold value most likely as a result of a marked inertia of the transducing pathway. Since the upregulation pattern of the transporters of the different osmolytes notably differs, we speculate that the activation threshold varies from one gene to another. PMID- 11008217 TI - Subcommissural organ/Reissner's fiber complex: characterization of SCO-spondin, a glycoprotein with potent activity on neurite outgrowth. AB - In the developing vertebrate nervous system, several proteins of the thrombospondin superfamily act on axonal pathfinding. By successive screening of a SCO-cDNA library, we have characterized a new member of this superfamily, which we call SCO-spondin. This extracellular matrix glycoprotein of 4,560 amino acids is expressed and secreted early in development by the subcommissural organ (SCO), an ependymal differentiation located in the roof of the Sylvian aqueduct. Furthermore, SCO-spondin makes part of Reissner's fiber (RF), a thread-like structure present in the central canal of the spinal cord. This novel protein shows a unique arrangement of several conserved domains, including 26 thrombospondin type 1 repeats (TSR), nine low-density lipoprotein receptor (LDLr) type A domains, two epidermal growth factor (EGF)-like domains, and N- and C terminal von Willebrand factor (vWF) cysteine-rich domains, all of which are potent sites of protein-protein interaction. Regarding the huge number of TSR, the putative function of SCO-spondin on axonal guidance is discussed in comparison with other developmental molecules of the CNS exhibiting TSR. To correlate SCO-spondin molecular feature and function, we tested the effect of oligopeptides, whose sequences include highly conserved amino acids of the consensus domains on a neuroblastoma cell line B 104. One of these peptides (WSGWSSCSRSCG) markedly increased neurite outgrowth of B 104 cells and this effect was dose dependent. Thus, SCO-spondin is a favorable substrate for neurite outgrowth and may participate in the posterior commissure formation and spinal cord differentiation during ontogenesis of the central nervous system. PMID- 11008218 TI - Targeted disruption of Muller cell metabolism induces photoreceptor dysmorphogenesis. AB - Within the retina, the Muller cells and photoreceptors are in close physical proximity and are metabolically coupled. It is unknown, however, whether Muller cells affect photoreceptor differentiation and outer segment membrane assembly. The objective of this study was to determine whether targeted disruption of Muller cell metabolism would induce photoreceptor dysmorphogenesis. Intact isolated Xenopus laevis embryonic eyes were cultured in medium with or without Muller cell-specific inhibitors (i.e., alpha-aminoadipic acid and fluorocitrate). To assess Muller cell injury, the gross retinal morphology was examined along with immunocytochemical assessment of Muller cell-specific protein expression patterns. The steady-state levels of opsin were quantified to determine whether the Muller cell inhibitors negatively affected photoreceptor protein synthesis. Muller and photoreceptor cell ultrastructure was scrutinized and the organization of the outer segment membranes was graded. In control retinas, there was no swelling of Muller cell cytoplasm. Glial fibrillary acidic protein (GFAP) was undetectable, whereas glutamine synthetase was abundant. The steady-state level of opsin was high and photoreceptors elaborated properly folded outer segments. Exposure to both Muller cell-specific inhibitors induced swelling of Muller cell endfeet, cytoplasmic paling and alterations of Muller cell-specific protein expression patterns. The steady-state level of opsin in retinas exposed to alpha aminoadipic acid was unchanged compared with control eyes, whereas, in eyes exposed to fluorocitrate, opsin levels were slightly reduced. The most significant finding was that targeted disruption of Muller cell metabolism adversely affected photoreceptor outer segment membrane assembly, causing dysmorphogenesis of nascent outer segments. These results suggest that the termination signal(s) necessary for proper outer segment folding were disrupted by targeted inhibition of Muller cells and support the hypothesis that Muller cells interact with photoreceptors through mechanisms that may regulate, at least in part, the assembly of photoreceptor outer segment membranes. PMID- 11008219 TI - Epidemiology of nosocomial candidiasis: the importance of molecular typing. AB - Modern epidemiology studies now require that nosocomial pathogens be characterized to the subspecies level whenever possible to better define infectious processes and modes of transmission. In general, if isolates are classified as different by at least one molecular typing method, they may be assumed to represent different strains and to reflect independent infections. If the isolates are the same, it may be assumed that cross infection has occurred or that the patients were infected by exposure to a common source. Typing methods may also be used to address clinical problems related to distinguishing reinfection versus relapse of an infection, and to examine the development of antifungal resistance among fungal pathogens during the course of antifungal therapy. Determining DNA fingerprints of sequential isolates from patients undergoing antifungal therapy has been useful in demonstrating the potential for the development of antifungal resistance in previously susceptible strains and for detecting the substitution of a more resistant strain for a more susceptible strain in the face of intense antimicrobial pressure. In order to be useful as an epidemiological typing method, a DNA fingerprinting system must effectively distinguish between genetically unrelated strains, identify the same strain in separate samples, and reflect genetic relatedness or unrelatedness (genetic distance) among strains. PMID- 11008220 TI - Candiduria in hospitalized patients: a review. AB - The presence of Candida species in the urine is frequent among hospitalized patients. It represents a major challenge to the physician because it is unclear whether candiduria represents colonization or infection, whether the bladder or the kidney is involved in infection, or whether it represents a surrogate marker for systemic infection. This picture is more complicated because there are few prospective studies addressing the issue of when and how to treat a patient with candiduria. Strategies for management are based on the presence or absence of other features such as anatomic genitourinary abnormalities, renal transplant, and fever. If treatment is considered, fluconazole is the drug of choice, provided the agent is not Candida krusei or Candida glabrata. PMID- 11008221 TI - Assessing efficacy by measuring CD(4) counts and quality of life of AIDS patients treated with ritonavir, AZT and 3TC. AB - Lamivudine and zidovudine are proving to be an important antiretroviral combination against HIV that is superior to monotherapy. Recently, with the appearance of protease inhibitors, ritonavir has been shown to be a powerful drug when used in combination with reverse transcriptase inhibitors. The objective of this study was to observe the efficacy, adverse events, and changes in the quality of life of AIDS patients receiving treatment for the first time using AZT, 3TC and ritonavir as combination therapy. We selected 36 patients diagnosed with AIDS due to opportunistic infections and evaluated them by assessing their score on quality of life scales (Karnofsky, uniscale - Quality of Life, and Quality of Life Scale), T CD(4) and CD(8) lymphocyte counts, bodyweight and symptoms during a 6 month period. Assessments were made at 2 month intervals. One patient was excluded from the trial, therefore, 35 were assessed during 6 months. RESULTS: Bodyweight increased an average of 7.2%, CD(4) increased 260 cells/mm(3) and CD(8) increased 198 cells/mm(3). The Karnofsky and uniscale QOL scales reached 100% on the fourth visit. The Quality of Life Scale showed an important increase during this study from 5.5+/-2.3 to 9.7+/-0.5. Adverse events were observed in 25.0% of the patients, most being slight. One patient had to stop taking ritonavir due to nausea and vomiting. We conclude that AZT, 3TC, and ritonavir restored the quality of life for the AIDS patients studied in terms of psychosocial aspects and overall health conditions during 6 months of treatment. The adverse events were probably related to ritonavir, but they were slight and disappeared after 2 weeks. There was a significant increase in the average number of CD(4) lymphocytes during 6 months of treatment. PMID- 11008222 TI - A prospective and randomized study using ribavirin as monotherapy for the treatment of naive patients with chronic hepatitis C. AB - In order to evaluate the response to ribavirin in previously untreated patients with chronic hepatitic C, 39 patients were selected for a double-blind prospective and randomized trial, and divided into two groups: ribavirin-group (19 patients) and placebo-group (20 patients). Ribavirin was administered orally for 24 weeks (600 mg/day, followed by 1,000 mg/day and 1,200 mg/day each one for 8 weeks). After 3 months of drug administration, the patients were evaluated by measuring biochemical, virologic and histologic responses. After this phase, ribavirin was offered to the patients who had received placebo (second phase). The results showed that the patients who received ribavirin showed a higher reduction in serum alanine aminotransferase (ALT) activity than patients in the placebo group. Among the patients in the ribavirin-group, a complete biochemical response (ALT levels normalized) was observed in 3 patients (16%), and a partial response (reduction greater than 50% of the initial value of ALT activity) in 4 (21%). In the 20 patients in the placebo group, only 1 showed a partial response (5%). In the second phase of the study, among 16 patients who received ribavirin, 4 (25%) showed a complete and 5 (31%) a partial biochemical response. HCV-RNA did not become negative in any patient during the two phases. A reduction in the score of portal and lobular activity was observed in patients who received ribavirin, but statistical analysis did not identify differences. This study showed that ribavirin alone induces a biochemical response (ALT reduction) in some patients with chronic hepatitis C, which may be associated with a reduction in hepatic inflammatory activity reduction, but the changes are not sufficient to recommend initial monotherapy with ribavirin. PMID- 11008223 TI - Hepatitis B and hepatitis C prevalence among blood donors and HIV-1 infected patients in Florianopolis--Brazil. AB - Information is scarce on the prevalence of hepatitis B (HBV) and hepatitis C (HCV) among voluntary blood donors and patients infected with the human immunodeficiency virus (HIV) in Florianopolis, Brazil. A total of 2,678 serum samples from 2,583 blood donors and 95 HIV-infected patients, collected between April, 1994, and March, 1995, were examined for markers of HBV and HCV. All the samples were analyzed to detect HBV and HCV markers (HBsAg, anti-HBc, and anti HCV). Hepatitis B and C prevalence among the studied blood donors reached 9.3% and 1.0%, respectively; 0.7% being seropositive for HBsAg and 9.2% for anti-HBc. It was also verified that 0.1% of blood donors were seropositive for HBsAg alone, 8.6% seropositive for the anti-HBc alone, and 0.6% presented a positive reaction for both of the HBV markers studied. Among HIV-infected patients, prevalence of 69.5% and 54.7% for hepatitis B and hepatitis C, respectively, were observed. Of these patients, 18.9% were seropositive for HBsAg, and 66.3% for the anti-HBc. The prevalence of a reaction for HBsAg alone, and for anti-HBc alone was 3.1% and 50.5%, respectively, for HIV-infected patients, whereas 15.8% were seropositive for both of the studied markers. HBV and HCV coinfection was 0.1% in blood donors, and 40% of those patients tested seropositive for HIV. Results show prevalence of HBV and HCV infection to be significantly greater among HIV infected patients than among blood donors. These observations confirm the high frequency of HIV-infected patients exposure to these other viruses. PMID- 11008224 TI - Tissue pharmacokinetics of amoxicillin. An experimental design in rats. AB - Amoxicillin is used as the drug of first choice in many situations in medicine and dentistry, in spite of several reports regarding bacterial resistance. There is little data about the tissue concentration of this antimicrobial agent. Serum levels of amoxicillin have been evaluated in detail, but tissue levels have not. This study was carried out to determine the tissue concentration of amoxicillin during the first 10 h after administration. Four polyurethane sponges were implanted in the backs of 54 male rats. After 14 days, they received 40 mg/kg of po amoxicillin suspension. The animals were killed in groups of 6 at 15, 30, 60, 90, 120, 240, 360, 480 and 600 min after the administration. Serum, placed on paper discs, and granulomatous tissue were assayed by a microbiological method using Mueller Hinton agar inoculated with 108 cfu of Staphylococcus aureus (ATCC 25923). After 18 h of incubation, the inhibition zones were measured. It was observed that the drug in the serum and the tissue reached higher concentrations than MIC and MBC within a period of 30 min and 8 h following administration. We conclude that this method can allow determination of antibiotic tissue concentration without the need for infecting the animal and, therefore, without the associated animal pain and suffering in presently used models. PMID- 11008225 TI - Kikuchi's disease: report of 2 cases and a brief review of the literature. AB - Kikuchi's lymphadenitis is a histiocytic necrotizing lymphadenitis without granulocytic infiltration, with fever and generally with a benign course, despite its pathologic resemblance to malignant lymphoma. The illness usually begins with localized cervical adenopathy in young adult females, predominantly before the fourth decade of life. Clinically, several agents could be the cause of such an illness--toxoplasmosis, herpes group viral infection, cat scratch disease, lymphoma, SLE and other infectious agents. The diagnosis is made by using a histological technique. The lymph node biopsy reveals fibrinoid necrosis, loss of lymph node structure with many histiocytes and an absence of granulomatous reaction. The immunohistochemical analysis shows that the main affected cellular components are the T cells. Laboratory exams show an erythrocyte sedimentation increase, neutropenia, leukopenia and lymphocytosis. We describe two cases that were followed since 1994 and 1996, respectively. Both were females under 25 years old, who developed a febrile disease with lymphadenopathy. Lymph node biopsies showed necrotizing lymphadenitis without granulocytic infiltrations. The patients had no evidence of other systemic diseases. Our objectives are to present a rare cause of febrile disease with enlargement of cervical lymph nodes, to review Kikuchi's disease, and to alert the medical community to this rare cause of fever and lymphadenopathy. PMID- 11008226 TI - Publication of presentations made at The Brazilian Congress of Infectious Diseases: a note of appreciation to the authors. PMID- 11008228 TI - Commentary on 'Colorectal Cancer' by rodriguez-bigas et al PMID- 11008227 TI - Colorectal cancer: how does it start? How does it metastasize? AB - Colorectal carcinogenesis is a multistep process with an apparently orderly progression from benign tissue to invasive malignancy and metastases. Yet at the genome level, a considerably more chaotic situation exists, with order arising through the process of natural selection in the midst of genomic instability. Major pathways for colorectal carcinogenesis begin with suppressor loss or acquisition of a mutator phenotype, but there are other pathways known and yet to be described. These pathways result in the natural selection of cells with unstable genomes leading to malignancy and metastases. PMID- 11008230 TI - Commentary on 'Primary Prevention' by friedlich and stern PMID- 11008229 TI - Primary prevention: what can you tell your patient? AB - Colorectal cancer, with its high incidence and significant morbidity and mortality, remains one of the leading causes of cancer-related deaths worldwide. Primary prevention of colorectal cancer may be more cost-effective and practical than secondary prevention. The primary prevention of colorectal cancer involves dietary and environmental modifications, chemoprevention, and in some cases prophylactic surgery. PMID- 11008231 TI - Average-risk screening: is public policy compatible with individual needs? AB - The issue of the appropriateness of population-based screening for colorectal cancer has been much debated in the literature. The purpose of screening is to find early-stage disease among average-risk members of the population so that timely treatment may be implemented and mortality reduced. The goal of public policies regarding screening should be to facilitate this result. The adequacy of current public policy for reduction of colorectal cancer mortality is the main focus of this article. PMID- 11008232 TI - Commentary on 'Average-risk Screening' by levin and hawley PMID- 11008234 TI - Commentary on 'Familial adenomatous polyposis, hereditary nonpolyposis colon cancer, and familial Risk' by thorson and faria PMID- 11008233 TI - Familial adenomatous polyposis, hereditary nonpolyposis colon cancer, and familial risk: what are the implications for the surgeon? AB - Recent advances in the understanding of the molecular biology of colorectal cancer have resulted in many new implications for surgeons. To continue providing sound patient care, surgeons must familiarize themselves with associated issues that include genetic counseling and its role in patient and family management. Issues related to genetic counseling are reviewed in this article. Recommendations for surgical therapy and surveillance methods are summarized for each of the hereditary syndromes. Failure to use these patient management tools in an effective way may be a source of future litigation. PMID- 11008235 TI - Staging: what makes sense? Can the pathologist help? AB - The staging of colorectal cancer continues to be an interesting and evolving process. Accurate staging can predict overall prognosis and helps to select appropriate treatment options. This article addresses the clinical and pathologic staging of colon and rectal cancer. The use of endorectal ultrasound in the preoperative clinical staging of rectal cancer is reviewed. The importance of surgical resection margins, lymph node retrieval rates, and tumor markers is discussed. PMID- 11008236 TI - Commentary on 'Staging' by bernick and wong PMID- 11008238 TI - Commentary on 'Current protocols and outcomes for colonic Cancer' by guleserian and bland PMID- 11008237 TI - Current protocols and outcomes for colonic cancer: why do we fail? AB - As the second leading cause of cancer-related deaths, colorectal cancer remains a significant health problem for American men and women. Advances in diagnosis, screening, surgical techniques, and adjuvant therapy have improved survival over the past 30 years. Although most novel therapies are in early stages of development, they offer potential for major advances in the management of colorectal cancer. Continued efforts focused on disease prevention, along with a better understanding of the tumor biology, cancer immunotherapy, and gene therapy will certainly yield more sophisticated and effective treatment strategies for all patients who are either at risk for or have already been diagnosed with colorectal cancer. As increasing numbers of clinicians, scientists, and health care professionals continue to address these issues, better interventions and therapies are likely to emerge. PMID- 11008240 TI - Commentary on 'Radical surgery for rectal Cancer' by cohen PMID- 11008239 TI - Radical surgery for rectal cancer: why we fail and rationale for current clinical trials of adjuvant therapy. AB - The major goals in the treatment of patients with rectal cancer are overall cure, maximizing local control, and maintaining quality of life. Quality of life issues include avoidance of a permanent stoma and maintaining adequate bowel, sexual, and urinary function. Optimizing the surgical technique with a more selective application of adjuvant therapy will meet these goals. This article describes the likely explanations for local and distant recurrence and delineates current clinical trials of adjuvant therapy directed toward minimizing failure despite potentially curative surgical resection. PMID- 11008241 TI - Current protocols and outcomes of local therapy for rectal cancer. AB - Rectal cancer is a devastating disease, with patients fearing the disease and the potential treatments that may alter sexual function, genitourinary function, and overall body image. Defining the precise role for less morbid approaches to this disease, such as local excision, is of critical importance in providing optimal care in the future. This article discusses endocavitary radiation and fulguration, local excision, and prospective studies. PMID- 11008242 TI - Commentary on 'Current protocols and outcomes of local therapy for rectal Cancer' by bleday and steele PMID- 11008244 TI - Commentary on 'Laparoscopic resection for colorectal Cancer' by lee et al PMID- 11008243 TI - Laparoscopic resection for colorectal cancer: is it justified? AB - Controversy remains regarding the appropriateness of laparoscopic methods for the curative resection of colonic neoplasms. Long-term results after minimally invasive resection must be shown to be equivalent or better than those after open resection in order to justify the new technique in the setting of cancer. This article discusses adequacy of resection and short-term results, long-term outcome data, port and abdominal wound tumors, oncologic and immunologic basic science data, and the role of laparoscopy in the treatment of rectal cancers. PMID- 11008245 TI - Operative techniques for radical surgery for rectal carcinoma: can surgeons improve outcomes? AB - Local recurrence rates following resection of rectal cancer with curative intent are extremely variable. An appropriate anatomic dissection of the rectum is essential to accomplish a complete removal of the tumor and has been described since the 1940s. In the last two decades, increased emphasis has been placed on the concept of total mesorectal excision as the fundamental technical step to minimize local recurrence rates. It is currently accepted, however, that complete removal of the mesorectum is not necessary for all rectal cancer cases. On the other hand, measurement of the free radial margins should be routinely performed to evaluate the local aggressiveness of the disease and assess the potential for a curative resection. PMID- 11008246 TI - Commentary on 'Operative techniques for radical surgery for rectal Carcinoma' by stocchi and wolff PMID- 11008248 TI - Commentary on 'Rectal cancer adjuvant therapy Controversies' by rakinic and Fry PMID- 11008247 TI - Rectal cancer adjuvant therapy controversies: When? What? Why? AB - Although the need for adjuvant therapy in high-risk rectal cancer is widely accepted, controversies continue regarding timing, mode, and agents employed. The current recommended practice and its scientific basis are reviewed. Studies of induction therapy are discussed. Evidence of efficacy of new anticancer agents and modes of drug delivery are presented. PMID- 11008249 TI - New adjuvant therapy for colon cancer: justified hope or commercial hype. AB - This article discusses the role of adjuvant chemotherapy for colon cancer. Six months of adjuvant 5-FU/leucovorin represent the standard of care for resected stage III colon cancer patients. The role of adjuvant chemotherapy for stage II patients remains controversial. Future directions in the adjuvant treatment of colon cancer include incorporation of newer agents such as irinotecan, oxaliplatin, oral fluorinated pyrimidines, and immunotherapeutic approaches. PMID- 11008250 TI - Commentary on 'New adjuvant therapy for colon Cancer' by galanis et al PMID- 11008251 TI - Follow-up: does it work? Can we afford it? AB - Follow-up of patients with colorectal cancer has been controversial. This is largely because studies have been small and inadequate to assess results. A recently performed meta-analysis confirms a very significant benefit to the follow-up of patients following curative resection of colorectal cancer. This article outlines the rationale and cost-effectiveness of a follow-up policy. PMID- 11008252 TI - Commentary on 'Follow-Up' by beart PMID- 11008253 TI - Decision-making: what tests to do? What choices to consider? AB - The treatment of recurrent colorectal cancer confronts the surgeon with a diagnostic and therapeutic challenge, particularly in cases involving the pelvis. Investigation and treatment in each case is tailored to the individual patient. Treatment is divided into those cases where an attempt at cure is possible and those where only palliation of symptoms is possible. This article seeks to help the surgeon confronted with this problem answer the following questions: Which patients with recurrent colorectal cancer are suitable for consideration of curative resection? What is the extent of disease recurrence? Is the disease resectable? The aim of this article is to offer advice in the logical and appropriate investigation and treatment of patients considered for curative or palliative therapy. PMID- 11008254 TI - Commentary on 'Decision-Making' by fazio and harris PMID- 11008255 TI - Current management of colorectal liver metastases. AB - Colorectal cancer metastatic to the liver represents an uncommon situation in surgical oncology in which metastasectomy can effect longevity and may lead to cure. Liver resection can be done safely and has drastically improved 5-year survival rates to upwards of 35%, but only a minority of patients is eligible. Advances in imaging techniques facilitate detection of metastases and improve patient selection. For unresectable patients, and as an adjunct to resection, local disease control may be achieved with cryosurgery, radiofrequency ablation, or regional chemotherapy delivered by way of infusion through the hepatic artery. Areas of ongoing investigation include regional gene therapy and interference with tumor growth by inhibition of angiogenesis. PMID- 11008256 TI - Commentary on 'Current management of colorectal liver Metastases' by martin and warren PMID- 11008257 TI - Pontocerebellar hypoplasia--how many types? AB - An original article describes a sibship with early fatal pontocerebellar hypoplasia of a yet unclassified type, with the accompanying features of polyhydramnios and neonatal myoclonus. Autopsy in one patient excluded spinal anterior horn involvement, which argues against pontocerebellar hypoplasia type I (PCH-1). The present PCH classification and literature are briefly reviewed. Four previous publications bear similarity to the present report. Definite classification as a genetically separate entity, however, remains elusive pending localization and identification of the gene(s) involved. PMID- 11008258 TI - What's new in congenital disorders of glycosylation? PMID- 11008259 TI - Provocation of non-convulsive status epilepticus by tiagabine in three adolescent patients. AB - Three girls, two aged 12 years and one aged 17 years with refractory localization related epilepsy were treated on an add-on basis with tiagabine. At dosages 22.5 30 mg/day (0.45-0.57 mg/kg/day) longstanding non-convulsive status epilepticus was noted in all three patients. The events of non-convulsive status epilepticus subsided following reduction in tiagabine dosages. In two cases, tiagabine was withdrawn, whereas it was continued at a lower dosage in one case. This is the first report of non-convulsive status epilepticus provoked by tiagabine in adolescent patients. PMID- 11008260 TI - Early fatal pontocerebellar hypoplasia in premature twin sisters. AB - We report clinical, neuroradiological and neuropathological findings of monozygotic twin sisters born at 30 weeks' gestation, with pontocerebellar hypoplasia (PCH) similar but not identical to type 2 PCH. They presented with hypertonia, jitteriness, spontaneous and provoked myoclonic jerks (hyperekplexia), apnoeic episodes, and progressive microcephaly. They died at 7 weeks of age from respiratory failure. PMID- 11008261 TI - Retinitis and dementia in a pregnant girl: an unusual case. AB - A 14-year-old pregnant Caucasian girl presented with a 1-week history of dementia. She had presented 1 year prior with acute unilateral visual impairment and was noted to have macular degeneration of presumed infective aetiology. This evolved to a pigmentary macular lesion. During the course of the current presentation she developed myoclonic jerks. An electroencephalogram revealed periodic spike and slow wave complexes at 1-2 second intervals. Blood and cerebrospinal fluid examination showed raised anti-measles IgG antibody titres. Intrathecal synthesis of anti-measles virus antibody was demonstrated unequivocally and a diagnosis of subacute sclerosing panencephalitis was made. A healthy male infant was delivered by elective caesarean section at 33 weeks' gestation. She continued to deteriorate clinically despite treatment with intraventricular alpha-interferon. She had not had primary immunization against measles. PMID- 11008262 TI - Mesenteric angina complicating a mesodermal anomaly. AB - A child with macrocephaly-cutis marmorata developed severe abdominal pain thought to represent mesenteric angina. There were abnormalities of the aortic and mesenteric vasculature not previously reported in this condition. Angina therapy afforded amelioration of his symptoms. Mesenteric angina should be considered as a cause for abdominal pain in children with mesodermal anomalies. PMID- 11008263 TI - Moyamoya syndrome with protein S deficiency. AB - Moyamoya disease is a cerebrovascular disease with progressive occlusion of both internal carotid arteries and of their branches and formation of a new vascular network at the base of the brain. Because of the angiographic appearance, it is named as moyamoya. The clinical features are cerebral ischaemia, recurrent transient ischaemic attacks, sensorimotor paralysis, convulsions and migraine like headaches. A 10-year-old child who acutely developed hemiparesis, weakness and aphasia was found to have moyamoya disease and heterozygous protein S deficiency. This case shows us that during the thromboembolic events the coexistence of protein S deficiency and moyamoya should be investigated. PMID- 11008265 TI - Forthcoming meetings PMID- 11008264 TI - Septo-optic dysplasia in combination with a pigmented skin lesion: a case report with nosological discussion. AB - In this case report a patient with bilateral optic nerve hypoplasia, schizencephaly and a pigmented skin lesion is described. The diagnosis of de Morsier syndrome or septo-optic dysplasia is put forward on the basis of the diagnosis of optic nerve hypoplasia. The differential diagnosis with Jadassohn's naevus phakomatosis is discussed. The importance of direct ophthalmoscopy of optic nerve abnormalities is stressed, as well as of magnetic resonance imaging, which has become a guideline in the classification of this syndrome. PMID- 11008266 TI - Advantages of rhythmic movements at resonance: minimal active degrees of freedom, minimal noise, and maximal predictability. AB - Using time delay embedding, the authors applied phase space reconstruction to the time series of rhythmic movements of a hand-held pendulum. Subjects (N = 6) produced the manual oscillations about the wrist at the pendulum's resonant frequency and at a higher and a lower frequency. The number of active degrees of freedom required to capture the dynamics of the rhythmic behavior was 3 for the resonant frequency and 4 for each of the nonresonant frequencies. The residual high-dimensional noise was similarly lowest for the resonant frequency. Whereas 33% and 20%, respectively, of the vectors in the phase spaces of the dynamics higher and lower than resonance were unpredictable, only 12% were unpredictable at resonance. Finally, the predictability of the evolving dynamics extended farther into the future for oscillations at the resonant frequency. At resonance, the prediction horizon was 5 times farther than the prediction horizon for the higher than resonance behavior and 2.5 times farther than that for the lower than resonance behavior. The results suggest that, in pendular oscillations of a limb or limb segment, attunement of the central nervous system to the resonant frequency minimizes the variables to be controlled and maximizes the predictability of the rhythmic movement's chaotic dynamics. PMID- 11008267 TI - Changes in movement symmetry associated with strengthening and fatigue of agonist and antagonist muscles. AB - The hypothesis that strengthening or fatiguing procedures applied on active muscles can affect the symmetry of rapid, discrete movements was tested. Subjects (N = 12) performed rapid, consecutive elbow flexions and extensions between 2 targets before and after (a) applying a strength training program, (b) fatiguing elbow flexors, and (c) fatiguing elbow extensors. The results demonstrated that an increase in strength of elbow extensors caused by applied strength training is associated with an increase in the symmetry ratio (i.e., acceleration time divided by deceleration time) of elbow flexion movements. The symmetry ratio also increased and decreased in movements when agonists and antagonists were fatigued, respectively. Because the strength training and fatiguing procedures are both known to affect muscle force, the data are interpreted as changes in muscles' ability to exert the force while acting as agonists or antagonists. Namely, muscles need equal impulses of force (torque multiplied by time) to accelerate and, thereafter, to decelerate the limb while performing a rapid, discrete movement. The symmetry ratio may therefore be changed so that more time will be provided for muscles that become relatively weaker (compared with their antagonists) because a strengthening or fatiguing procedure has been applied, whereas a shorter time period should be sufficient for action of their stronger antagonists. Although, in the literature, the studied phenomenon has been discussed as a predominantly motor control phenomenon, the present data suggest that the movement symmetry could also be related to agonists' and antagonists' ability to exert force, particularly while performing rapid, discrete movements. PMID- 11008268 TI - Visual dominance in amending the directional parameter of feedforward control. AB - The authors examined visual dominance between trials in which the movement program was amended (i.e., off-line processing). Weighting between visual and proprioceptive feedback was examined in a trial-by-trial analysis of the directional parameter of feedforward control. Eight participants moved a cursor to a target displayed on a computer screen by manipulating a hand-held stylus on a digitizing tablet. In the first 30 trials, the cursor followed the stylus movement (practice condition). In the next 30 trials, the directional error of the stylus movement was presented in the opposite direction (reversal condition). Subjects knew the presence and the nature of the reversal. In the last 10 trials, the reversal was withdrawn (transfer condition). Directional error of feedforward control was relatively small in the practice condition, and it increased gradually in 1 of 2 directions as trials proceeded in the reversal condition. Positive aftereffect was observed in the transfer condition. A constant increment of the directional error indicated that both visual and proprioceptive feedback are registered, with higher weight on vision, and that weighting between those inputs is determined automatically or is fixed without any strategic control. PMID- 11008269 TI - Physical and observational practice afford unique learning opportunities. AB - In 2 experiments, the authors studied the effectiveness of physical and observational practice on learning and the effect on learning of combining physical practice and observation, as compared with providing physical practice alone. In Experiment 1, retention and transfer performance of 30 university students after physical, observational, or no practice were contrasted. Consistent with findings from other studies, the retention results indicated that observational practice is inferior to physical practice. The transfer data indicated no differences between observation and physical practice groups. In Experiment 2, retention and transfer performance of 30 participants in physical and combined (alternating physical and observational) practice groups were contrasted. The retention results showed no differences between the combined and physical practice groups, but the combined group performed significantly better than the physical practice group on the transfer test. Those findings suggest that a combination of observation and physical practice permits unique opportunities for learning beyond those available via either practice regimen alone. PMID- 11008270 TI - A perception-action coupling type of control in long jumping. AB - The authors' goal was to identify the control mechanisms used by long jumpers (N = 6) to precisely position their foot at the board. In addition to the intertrial method usually used in previous research, an original method based on a trial-by trial analysis was also implemented. If the approach to the board in long jumping encompasses two distinct sequences separated by a key step that marks the initiation of visual control, then a trial-by-trial analysis should reveal those sequences, regardless of the amount of adjustment: The step number at which regulation is initiated should be the same irrespective of the amount of adjustment. If, in contrast, a perception-action coupling mechanism operates, then the step number at which regulation is initiated should be a function of the amount of adjustment: A linear relation between those 2 variables should emerge. The results of the present study are compatible with continuous control mechanisms based on a perception-action coupling. PMID- 11008271 TI - Effect of sex and joystick experience on pursuit tracking in adults. AB - Using a joystick, adults (n = 39 males, 40 females in Experiment 1; n = 35 males, 40 females in Experiment 2; and n = 18 males, 18 females in Experiment 3) performed a computerized pursuit tracking task. Contrary to previously reported findings, the males were not more accurate than the females when performance was adjusted for prior perceptual-motor experience. Although no sex differences were found in a speeded tracking task, in an inverted tracking task the males exhibited a significant performance advantage; that advantage remained after several blocks of practice. Because participants' performance was adjusted statistically for prior perceptual-motor experience, the male advantage in inverted tracking was not related to experience. Rather, more proficient inverted tracking performance was associated with higher 3-dimensional mental rotations scores. In sum, sex differences in normal pursuit tracking may be better explained by differences in perceptual-motor experience. Inverted tracking, however, may depend on proficiency with spatial transformations. PMID- 11008272 TI - Bernstein's theory of movement behavior: historical development and contemporary relevance. AB - In present-day movement science, N. A. Bernstein's formulation of the problems of motor control is often taken as the starting point. The reliance on Bernstein has not brought agreement among his followers, however. In this article, the authors pose the following question: Does the disagreement arise from the structure of his work itself or from incomplete exploitation of his thinking? By using, inter alia, Bernstein's 24 English and German articles, the authors present an analysis of the development of Bernstein's theory of movement behavior, against the backdrop of the scientific progress in the Soviet Union in Bernstein's time and the clashes between Soviet politics and science. Bernstein addressed in his early articles the measurement and biomechanical analysis of movements. His experimental data soon indicated the need for a new understanding of the organization of movements, which he formulated in terms of coordination. Because of political problems, his work was interrupted; but after being "rehabilitated" and again allowed to work, Bernstein aimed to explain how animals find and optimize the solutions to motor problems. The structure of the theory that ensued was comprehensive exactly by virtue of his repeatedly shifting focus between the different aspects of the organization of movement: More important than the answers he gave were the questions he asked. Moreover, the way he approached those questions may help scientists solve pressing problems in present-day movement science. PMID- 11008273 TI - Coordination of grip configurations as a function of force output. AB - In this investigation, the authors examined the coordination and control of force production by the digits of the hand as a function of criterion force level and grip configuration. Each adult participant (N = 6: 3 men and 3 women) was required to place the thumb and a finger (or fingers) upon load cells that were fixed to a grasping apparatus that was clamped to a table. In the task, participants had to match a criterion continuous constant total force level displayed on a computer screen. There were 10 trials at each grip configuration and criterion force level combination on each of 3 consecutive days. The results showed that (a) different grip configurations minimized error at each force level; (b) there was a specific digit pairing within a given grip configuration that produced the highest correlation of force output; (c) the correlation between the force output of digits generally increased at higher force levels; (d) error was reduced at each force level and grip configuration over the practice period; and (e) the organization of the force output of each digit varied as a function of digit, force level, grip configuration, and practice. The findings are consistent with the hypothesis that coordination of the digits in prehension is reflective of an adaptive, task-specific solution that is modified with practice. PMID- 11008274 TI - Prism exposure aftereffects and direct effects for different movement and feedback times. AB - The effects of movement time and time to visual feedback (feedback time) on prism exposure aftereffects and direct effects were studied. In Experiment 1, the participants' (N = 60) pointing limb became visible early in the movement (.2-s feedback time), and eye-head aftereffects increased with increasing movement time (.5 to 3.0 s), but larger hand-head aftereffects showed little change. Direct effects (terminal error during exposure) showed near-perfect compensation for the prismatic displacement (11.4 diopters) when movement time was short but decreasing compensation with longer movement times. In Experiment 2, participants' (N = 48) eye-head aftereffects increased and their larger hand-head aftereffects decreased with increasing movement time (2.0 and 3.0 s), especially when feedback time increased (.25 and 1.5 s). Direct effects showed increasing overcompensation for longer movement and feedback times. Those results suggest that aftereffects and direct effects measure distinct adaptive processes, namely, spatial realignment and strategic control, respectively. Differences in movement and feedback times evoke different eye-hand coordination strategies and consequent direct effects. Realignment aftereffects also depend upon the coordination strategy deployed, but not all strategies support realignment. Moreover, realignment is transparent to strategic control and, when added to strategic correction, may produce nonadaptive performance. PMID- 11008275 TI - The development of compensatory stepping skills in children. AB - The development of the ability to use the step for balance recovery was studied among twenty-five 9- to 19-month-old children. The children were grouped according to walking experience (4 levels) and exposed to backward support surface translations, 8 cm in amplitude, under 3 velocity conditions: 15, 20, and 25 cm/s. New walkers (up to 2 weeks' walking experience) used the step infrequently and ineffectively in response to threats to balance. Intermediate walkers (1-3 months' walking experience) showed an increasing use of the step and significant improvement in step execution compared with new walkers. Advanced walkers (>3; months' walking experience) experienced no falls throughout the protocol, capturing balance with feet-in-place or step responses under all perturbation conditions. A significant developmental transition in the emergence of the compensatory step occurred between the new walker and the intermediate walker experience levels, that is, within the first 3 months of walking experience. Three to 6 months' experience was required for the development of an effective stepping response. A concomitant change in mediolateral stability paralleled the emergence of compensatory stepping. PMID- 11008276 TI - [Effect strength variation in the single group pre-post study design: a critical review]. AB - In Germany, studies in rehabilitation research--in particular evaluation studies and examinations of quality of outcome--have so far mostly been executed according to the uncontrolled one-group pre-post design. Assessment of outcome is usually made by comparing the pre- and post-treatment means of the outcome variables. The pre-post differences are checked, and in case of significance, the results are increasingly presented in form of effect sizes. For this reason, this contribution presents different effect size indices used for the one-group pre post design--in spite of fundamental doubts which exist in relation to that design due to its limited internal validity. The numerator concerning all effect size indices of the one-group pre-post design is defined as difference between the pre- and post-treatment means, whereas there are different possibilities and recommendations with regard to the denominator and hence the standard deviation that serves as the basis for standardizing the difference of the means. Used above all are standardization oriented towards the standard deviation of the pre treatment scores, standardization oriented towards the pooled standard deviation of the pre- and post-treatment scores, and standardization oriented towards the standard deviation of the pre-post differences. Two examples are given to demonstrate that the different modes of calculating effect size indices in the one-group pre-post design may lead to very different outcome patterns. Additionally, it is pointed out that effect sizes from the uncontrolled one-group pre-post design generally tend to be higher than effect sizes from studies conducted with control groups. Finally, the pros and cons of the different effect size indices are discussed and recommendations are given. PMID- 11008277 TI - [Clinical studies in rehabilitation research--problems and solutions from biometrical perspective]. AB - Methodology of clinical studies is highly sophisticated in drug research. But clinical trials are also necessary to demonstrate efficacy and safety of rehabilitation treatment. The call for evidence based medicine has also reached rehabilitation. However, in rehabilitation medicine it is much more difficult to design and conduct clinical trials with a high methodological standard. Among the reasons are: A comparable control group is necessary because spontaneous healing and unspecific measures contribute to therapeutic success, too. But what could "placebo rehabilitation" look like? The masking of therapies (blinded studies) will hardly ever be possible. Therefore, it is more difficult to achieve the same treatment and observation for the treatment and control group. Treatments in rehabilitation take longer to become effective than a drug and maybe the success will disappear after some time. Therefore, long-term trials and follow-ups are necessary. Such studies are expensive, need a strong organisation, and drop-outs are unavoidable. An appropriate outcome variable does not always exist. "Return to work" is an important, reliable and valid variable, but it delivers only one bit of information per patient. As a consequence, smaller progress in rehabilitation can only be demonstrated with large sample sizes. Outcome variables based on time enable studies with reasonable sample sizes. Sometimes it is more difficult to obtain acceptance of randomisation in rehabilitation patients than in acute patients. Some rehabilitation hospitals have only recently begun to take an interest in controlled clinical trials, hence are not so experienced. Nevertheless, controlled clinical trials delivering convincing results are possible in rehabilitation medicine as well. But biometrical consultation is necessary e.g. for study design, study conduction and evaluation. Most important points are the methodology of the study design and its practicability. Especially in these topics rehabilitation physicians and biometrician have to cooperate. PMID- 11008278 TI - [Patient expectations regarding methods and outcomes of their rehabilitation--a controlled study of back pain- and cancer patients]. AB - Although patient expectations are important factors of the success of rehabilitation, they have not yet received much attention in research. In the present cross-sectional study, n = 248 rehabilitation patients, n = 160 suffering from chronic back pain and n = 88 suffering from oncological diseases (breast cancer, cervical cancer, ovarian cancer, colorectal cancer), were assessed at the time of admission to a rehabilitation clinic, using a newly developed self-report questionnaire to evaluate their expectations regarding the process and outcome of their rehabilitation. Results show on an item level that non-specific process expectations such as balneophysical treatments or features of the surroundings of the clinic were most prevailing. Across both diagnostic groups, common goals of rehabilitation such as reduction of complaints, physical fitness and recreation scored highest. On the basis of factor analysis, 10 scales of process expectations were constructed: medical attention, physical training, vocational counselling, balneophysical treatment, health information, stress management training, support groups, relief from the strains of everyday life, alternative medicine, and pleasant surroundings. Nine dimensions of outcome expectations were detected: ability for work, physical fitness, reduction of complaints, behaviour change, reduction of body weight, positive body feeling, vitality, enjoyment of life, and social contact. In the univariate analysis, orthopaedical patients scored higher on balneophysical treatments and vocational counselling (process expectations) as well as reduction of complaints and ability for work (outcome expectations). However, there were effects of age, gender and working status, too. Thus these differences between diagnostic groups turned out to be nonsignificant after controlling for the effects of age, gender, and working status. On the other hand, some effects of the medical condition had been hidden in the univariate comparison and were only revealed when adjusting for the moderator variables. Those differences show that oncological patients had higher expectations than back pain patients. Patients' expectations were correlated with functional status and, to a lesser degree, emotional distress and locus of control. To conclude, our study demonstrated that medical condition, sociodemographic factors, working status and functional capacity are important determinants of patient expectations regarding the process and outcome of rehabilitation. PMID- 11008279 TI - [High-mountain climate therapy for skin diseases and allergies-- mode of action, therapeutic results, and immunologic effects]. AB - Dermatological-allergologic climatotherapy is interpreted as a therapy within a specific climate with proven therapeutic benefits, immediate and longterm. Intensive classical dermatological in-patient therapy is combined with specific climatic effects. Primarily, the climate of the high mountains (1560 m) and of the North Sea islands is of proven efficacy for dermatoses and allergic diseases such as atopic dermatitis (neurodermatitis), eczema, psoriasis, T-cell lymphoma, bronchial asthma. Specialized therapeutic utilities exist. Directly influencing climatic factors such as insolation, thermic-hygric and aerosol conditions without or with diminished allergic potency and nonspecific stimulating climate factors change immune functions and effect stabilization. The therapeutic immediate and longterm efficacy of the high mountain climate is proven by excellent follow-up results. Its superiority to the dermatological therapy applied at home is evident. Measurement and analysis of climate efficacy has however proven difficult because of its complexity. The findings of several recent clinical and biochemical studies are presented. PMID- 11008280 TI - [Causes and correction of abnormal gait patterns due to prosthesis in above-knee amputees]. AB - Abnormal gait patterns cause an increase in the energy cost of walking in above knee amputees. Disturbances of the walking pattern are often caused both by the amputated patient himself and by incorrect prosthetic fitting. The early detection and correction of causative factors is of great importance for successful rehabilitation of these amputees. During the follow-up examinations the prosthesis must be inspected for a correct fitting and individually appropriate knee stabilizing components. Prior to any corrective measures it must be excluded that the artificial limb has been put on incorrectly by the amputee himself. PMID- 11008281 TI - [Rehabilitation networking illustrated by case management after occupational accidents]. AB - No net without a spider! After work accidents the victims can rely on the infrastructure created by the German accident insurance scheme (Berufsgenossenschaften). A surgeon, a so-called transit physician, is responsible after acute treatments also for all phases of rehabilitation. A Case Manager (Berufshelfer), an employee of the Berufsgenossenschaften, advises the victim in all matters, including vocational rehabilitation and personal concerns. Both factors combine to form a network that orientates on the social needs of the injured worker, on the financial tasks of the insurance agency and on incentives to the rehabilitation facilities. The goals comprise "rehabilitation before compensation" and "job retention instead of job loss". PMID- 11008282 TI - [Report on the symposium "Patient education--perspectives from patient-oriented medicine" Bremen, Jan. 20-21, 2000]. PMID- 11008283 TI - [Workshop on "Work-related procedures within the framework of medical rehabilitation" Wurzburg, Jan. 25-6, 2000]. PMID- 11008284 TI - [Cerebral seizures in neonatal period: semiology, evolution and factors of influence]. AB - INTRODUCTION: In spite of their incidence being much greater than at any other period during life, seizures occurring during the neonatal period are difficult to identify, since their patterns are not well organized on account of insufficient anatomical, physiological and/or biochemical development. OBJECTIVE: To analyse the semiology of seizures during the neonatal period, their classification and subsequent evolution. PATIENTS AND METHODS: We made a retrospective study of 60 patients who had convulsions during the neonatal period, selected from amongst the children admitted to the Neonatology Department of the Hospital Infantil Universitario Virgen de Rocio in Sevilla, between 1990 and 1998. We investigated 22 medical variables related to the clinical history, neurological examination, neuroimaging studies, EEG and drugs used. RESULTS: Anomalies were found on the initial examination in 83.3% of the cases. The hypoxic-ischemic syndrome was the commonest etiology, followed by hemorrhages, metabolic disorders, cardiopathies, malformations and infectious diseases. From the semiological angle, the seizures were seen as partial or generalised increase in muscle tone, followed by clonias, hypotonias and subtle seizures. Cerebral angiography was the most commonly used diagnostic imaging technique. There was a statistically significant relationship between the appearance of sequelae and the earliness of the occurrence of the seizures, but not with the duration, semiology or frequency. CONCLUSIONS: Seizures in the newborn are habitually seen in the hypoxic-ischemic syndrome, and usually present as alterations in muscle tone. Phenobarbitone is still the drug of choice for the treatment of seizures in the neonatal period. PMID- 11008285 TI - [Type I neurofibromatosis presenting as a progressive cervical myelopathy. The first case reported in Kaxinawa Indians]. AB - INTRODUCTION: Type I neurofibromatosis is an autosomal dominant disorder with an estimated prevalence of 1/3,000. Half of the new cases are due to mutations; their penetration is complete in adults and may affect any ethnic group. We describe the first reported case of type I neurofibromatosis in a Kaxinawa Indian from the state of Acre, Brazil, in whom the presenting symptoms were of progressive tetraparesis due to multiple paraspinal neurofibromata. CLINICAL CASE: A 16 year old indigenous Kaxinawa boy presented with progressive myelopathy for the past six months. On neurological examination he had hyperreflexive spastic tetraparesia, predominantly on the right side, with the sensory level at C3/C4, multiple medullary automatisms and respiratory difficulty. He also had diffuse cafe-au-lait stains, including one particularly prominent one measuring 15 cm on his right buttock. MR of the spine showed many hypo-intense lesions at T1 and hyperintense lesions at T2, which took up contrast and were present at all the conjugation foramens bilaterally. These lesions originated at the roots and were suggestive of neurofibromata. On mediastinal tomography there were several neurofibromata in the mediastinum. The patient had a surgical operation for spinal decompression, with a laminectomy at C1/C2 and removal of the neurofibromata on the spinal cord at C1 and C2. The patient has a sister with multiple cafe-au-lait stains. CONCLUSIONS: Because of the traditional isolation of the Kaxinawa Indians, with whom contact was made at the beginning of the twentieth century, this first case reported of type I neurofibromatosis may be due to a new mutation which presented as a florid form of the disease with multiple spinal neurofibromata. PMID- 11008286 TI - [Cerebral arteritis and psychic involution in children. A report of one case with a good response to treatment]. AB - OBJECTIVE: To show the importance of the cerebral arteritis as etiology of the language and the intellectual involution in children. CLINICAL CASE: A boy started to show psychic and language involution since 18 months of life to arrive to an autistic behavior. After showing normal results in all the studies performed in order to investigate the possible etiologies, cerebral arteriography was performed. Cerebral arteritis affecting especially the right opercular artery was disclosed. Oral nicardipine administration was follow-up of a complete recuperation. CONCLUSIONS: Cerebral arteritis is very seldom managed as the cause of intellectual and/or language involution in children as it also occurs with the syndrome of acquired aphasia. However, this pathology has a good response not only to corticoids but also to calcium channel antagonists as it occurred in our patient. PMID- 11008287 TI - [Risk factors for ischemic stroke. I. Conventional risk factors]. AB - INTRODUCTION: Stroke, together with ischemic cardiopathy, may be considered to be a vascular epidemic in the developed world, where it is a major social and health problem. In Spain it is the commonest cause of death in women and the second commonest in men. DEVELOPMENT AND CONCLUSIONS: In this article we review the risk factors which cannot be modified (age, sex, inheritance, race, geography and climate) and the well-established risk factors which can be modified (arterial hypertension, cardiopathies with a high risk of embolization, markers for atherosclerosis, diabetes mellitus and transient ischemic accidents). A wide understanding of these risk factors permits the identification of persons with a high risk of having an ischemic stroke. Therefore preventive and therapeutic measures to reduce individual risk from strokes (and their recurrence) and hence reduce morbimortality from this, can be designed. PMID- 11008288 TI - [Diseases of mitochondrial DNA]. AB - INTRODUCTION: Human diseases caused by disorders of the mitochondrial metabolism have been described more than 30 years ago. Some of these are associated to defects in the oxidative phosphorylation system (OXPHOS system), the final pathway of the mitochondrial energetic metabolism, that leads to the synthesis of ATP. DEVELOPMENT: Part of the polypeptide subunits involved in the OXPHOS system are codified by the mitochondrial DNA (mtDNA). In the last 12 years, mutations (point mutations or deletions) in the mtDNA have been described and associated to well defined clinical syndromes caused by defects in the OXPHOS system. The clinical features of these diseases are very heterogeneous affecting in most cases to a great variety of organs and tissues. CONCLUSIONS: The correct diagnosis of these mitochondrial disorders require precise clinical, morphological, biochemical, and genetic data. The rapid advances in genetic analysis allow the rapid detection of mutations, even before the obtention of other type of analysis. PMID- 11008289 TI - [Cysteine proteinase and neurodegeneration]. AB - OBJECTIVE: This is a review of the part played by the cysteine proteases in different physiological and pathological processes. DEVELOPMENT: Apoptotic processes have a crucial function in control of the number of cells in multicellular organisms, both during development and throughout life. Alterations in these are closely related to different pathological processes, from cancer (with fewer apoptotic processes) to the degenerative disorders in which apoptosis is increased. Although the stimuli which may induce apoptosis are very varied, the apoptotic phenotypes are similar. Different metabolic routes are involved in apoptosis and in these changes, both in transcription and postranscription. The latter form the basis of this paper. We review the role of the cysteine protease family, in which the caspases and calpains are the best representatives, which have been related to different degenerative models. In this review we describe the stimuli and cascades of intracellular signalling which occur on activation. CONCLUSION: These proteases are involved in many situations involving the development and maintenance of the number of cells in the tissues, both physiological and pathological. They may be considered to be possible therapeutic targets in neurodegenerative diseases such as Alzheimer's disease, amyotrophic lateral sclerosis, Parkinson's disease and Huntington's chorea. PMID- 11008290 TI - [Practical considerations regarding the diagnosis and treatment of disorders of movement in childhood]. AB - OBJECTIVE: Disorders of movement in children frequently cause difficulties with both the semiology and the diagnostic approach. DEVELOPMENT: Based on our experience, we analyze useful strategies to overcome these difficulties and consider the clinical semiology, use of complementary tests and therapeutic approach. CONCLUSION: Correct clinical identification is the essential basis for differential diagnosis and therapeutic management of the disorders of movement. PMID- 11008291 TI - [Gilles de la Tourette syndrome]. AB - OBJECTIVE: To make a specific review of the definition and symptomatology of Gilles de la Tourette syndrome. DEVELOPMENT AND CONCLUSION: We made an basic syndrome evaluation and at the same time established a relationship with/between the most significant co-morbid features. We also refer briefly to current drug treatment and dosage and mention some of the most relevant side-effects of these drugs. The exact etiology of this disorder is still not fully understood but it is thought to be due to autosomal dominant inheritance, with marked penetration in males. PMID- 11008292 TI - [Does the cerebellum play a part in cognitive processes?]. AB - OBJECTIVE AND DEVELOPMENT: We review the different cognitive and behaviour disorders in whose genesis the cerebellum has been thought to play a part. These disorders include infantile autism, Williams' syndrome, attention-deficit hyperactivity disorder and congenital cerebellar hypoplasia. We also consider the consequences of an acquired cerebellar lesion in the light of the following neuropsychological sequelas. It would seem that the cerebellum is involved in the functioning of the executive system due to its relationship with the prefrontal operative system and in the function of the working memory, and may share some of the characteristics of the acquired frontal syndrome. PMID- 11008293 TI - [Right hemisphere syndrome in children: functional and maturity correlation of non-verbal learning disabilities]. AB - INTRODUCTION: This review presents the maturational model of the right hemisphere syndrome in children, its clinical manifestations and its relationship with non verbal learning disabilities. DEVELOPMENT AND CONCLUSIONS: The white matter model proposed by Rourke is presented and the clinical symptoms that are generally associated with right hemisphere dysfunction, independent of which hemisphere is affected, are highlighted. It has been proposed that the right hemisphere have proportionally more white matter than the left hemisphere which implies different adaptation processes for each one after brain lesions. There are different factors that determine these results: brain plasticity acts in different ways in each hemisphere; early brain injuries, affect specially right hemispheric functioning in its abilities to be in charge of new material processing and building new schemes that are also used by the left hemisphere. Some pathologies that are related with the clinical manifestations of non-verbal learning disabilities are presented. According with this model, they display basic neuropsychological features, even though their clinical manifestations could change between them. The Attention Deficit Hyperactivity disorder is specially considered as the most recent evidence of right hemispheric dysfunction, although results in this pathology are controversial, from the neuropsychological point of view. In general, the right hemisphere lesion in children has different implications when it is compared with adult's lesions. The white matter model could explain different processes in the plasticity brain mechanism. Preservation of basic language aspects is observed, but a deficit in non-verbal abilities that interferes with normal functioning is a rule. A specific clinical profile of assets and deficits is presented in most of the cases. These characteristics should be considered in treatment implementations. PMID- 11008294 TI - [Controversial or arguable therapies in neurodevelopmental disorders]. AB - OBJECTIVE: To analyze the therapies and methods not scientifically documented that more frequently are offer and used for the treatment of neurodevelopmental disorders. DEVELOPMENT: These therapies are divided into three main groups: a) Therapies directed upon brain functioning; b) Therapies directed upon nutritional needs, and c) Others. Parents and close relatives of children with developmental disorders are vulnerable to any person, institution or method that offers a quick and easy solution to their problem. It is a priority that all health professionals are familiar with the unproven therapies, that are offered for the therapy of developmental disorders, so that they can inform, educate and advise correctly to parents and close relatives of their patients. To deal with unproven treatments involves more than analyzing the available scientific data. It is required a comprehension and understanding of the personal and family dynamics in front of a threat of illness or disfunction. CONCLUSION: Responsible and well informed parents in relation to an unproven therapy should be free and have the right to decide whether or not to use a controversial procedure prior to a scientific determination of its validity; they also must keep in mind that at least some procedures, beside of not been useful, could be harmful for the physical, emotional and economical well-being of the patient and/or his family. PMID- 11008295 TI - [Risks and benefits of new antiepileptic agents in children]. AB - INTRODUCTION AND DEVELOPMENT: New antiepileptic drugs (AED) have generally a good pharmacokinetic profile. Their mode of action remains imperfectly known. Gabapentin (GBP), oxcarbazepine (OCBZ), topiramate (TPM), vigabatrin (VGB) and tiagabine (TGB) are mostly effective for partial seizures; lamotrigine (LTG) is efficacious for both partial and generalized seizures, it is also active in treatment of Lennox-Gastaut syndrome for which felbamate (FBM) is also effective. West syndrome often responds to VGB. The tolerance of the new AED is generally satisfactory but some serious, late side-effects have become apparent. Because of such possible toxicity, of the cost and of the remaining uncertainties about indications, the new AED are used mostly as add-on therapy for epilepsies resistant to conventional AED or as secondary monotherapy following withdrawal of other drugs used as monotherapy. Some new AED (LTG, GBP, TPM, OCBZ) have proved equal or sometimes superior to conventional agents. CONCLUSION: They may be especially useful in the treatment of certain resistant epilepsy syndromes such as Lennox-Gastaut and West syndromes for which conventional drugs often fail. PMID- 11008296 TI - [Epilepsy and learning disorders]. AB - INTRODUCTION: Children with epilepsy have more behavioral and cognitive problem than children with other chronic illness and children in the general population. Risk factors are multiple probably involving a combination of seizure, family and child variable, behavioral problems (depression, occur frequently but may be unrecognized). Anxiety disorders and psychoses are less common. DEVELOPMENT: The learning and behavioral found in epileptic children is multifactorial, first the effects of the conditions causing the epilepsy, in addition the epilepsy itself may cause, change in the nervous system through the metabolic and excitatory effects of the seizure and the neurotransmitter elements involved at the membrane level. The effects of the treatment modality, be it pharmacology or surgical may also play havoc with cognition function. CONCLUSION: The neurobehavioral effects of antiepileptic drug although probably less important with the newer agent than in the past, must be considered especially in polypharmacy as recent report have in dictated. PMID- 11008297 TI - [Atypical evolutions of benign partial epilepsy of infancy with centro-temporal spikes]. AB - OBJECTIVES: To stress that different atypical evolutions occur in the course of disease in some patients with the so called benign partial epilepsies of childhood (BPEC). To promote interest in finding clinical and/or electroencephalographic clues to recognize which patients might be prone to present these risks. METHODS: Follow up of 26 pediatric patients who started with typical clinical and EEG features of benign childhood epilepsy with centro temporal spikes (BCECTS) but presented reversible or persistent serious epileptic events including status epilepticus and language, cognitive or behavioral impairments. Repeated neurological examinations, EEG records and neuropsychological evaluations were done in the course of up to 14 years. Brain imaging studies (CAT and/or MRI) were obtained in all patients. RESULTS: Four groups of patients were recognized: 11 children with atypical benign partial epilepsy of childhood (ABPEC) followed during 4 to 13 years. All have finally recovered and attend normal schools, although five have learning difficulties. Three patients met diagnostic criteria for Landau-Kleffner syndrome (LKS). Two of them recovered from aphasia and in one, some language difficulties persist. Seven children showed status epilepticus of BCECTS but all of them are now normal after 3 to 14 years of follow-up. Five children showed mixed features of the three groups mentioned above and three of them fulfilled the criteria for diagnosis of the syndrome of continuous spike-wave during slow sleep (CSWS). CONCLUSIONS: A small proportion of cases starting with BCECTS evolve into either ABPEC, LKS, status of BCECTS or the syndrome of CSWS. Then, BCECTS is not always benign. Clinical and EEG markers should be sought to predict these atypical evolutions. PMID- 11008298 TI - [Staphylococcus aureus-related meningitis]. PMID- 11008299 TI - [Transient bilateral paralysis of abducens nerve in a newborn. A case report]. PMID- 11008300 TI - [Fukuyama type congenital muscular dystrophy (cerebro-muscular disease). A case report]. PMID- 11008301 TI - [Purple glove syndrome]. PMID- 11008302 TI - [HIV superinfection: myth or reality?]. AB - Most viral infections result in lifelong immunity against the virus, which prevents subsequent superinfection. In the light of recent findings on recombination of different viral clades, infection with two different HIV strains is unlikely to occur during primary infection. However, it is unclear whether true superinfection occurs during chronic HIV infection. If it does occur, superinfection could constitute a risk for HIV-concordant couples, since transmission of drug-resistant or more virulent viruses may result in worsening of the disease or treatment failure. We review the available data on superinfection and conclude that HIV-discordant couples should be informed of the theoretical risk of superinfection, especially where only one partner is receiving effective antiretroviral treatment. PMID- 11008303 TI - Intrathoracic tumours in von Recklinghausen's neurofibromatosis. AB - We review our cases of a rare pathology, intrathoracic tumours in von Recklinghausen's neurofibromatosis, involving 6 patients operated upon in our Surgery Department for intrathoracic neurogenic tumors. All had had a positive history of von Recklinghausen's neurofibromatosis prior to thoracic surgery. The management of this rare condition is discussed. We describe the presentation of the symptoms, the operative technique and the long-term outcome, when obtainable. Our results are compared with those of other authors, and the management of these cases is reconsidered. In conclusion, we recommend that thoracic surgery should only be considered when the patient becomes symptomatic, since malignant changes in the tumour are possible due to operative trauma. PMID- 11008304 TI - [Frequency of HFE gene mutations and genotype-phenotype correlations in patients with hereditary hemochromatosis in Switzerland]. AB - Hereditary haemochromatosis is one of the most common genetic disorders affecting populations of European ancestry. Isolation of a strong candidate gene, the HFE gene, allows genetic diagnosis in a large number of cases. However, different mutation frequencies have been reported in hereditary haemochromatosis patient populations from various geographic regions. Such data and phenotype-genotype correlations from Swiss patients with hereditary haemochromatosis are lacking. The objective of our study was to determine the frequency of HFE gene mutations in Swiss patients with hereditary haemochromatosis and to describe the clinical phenotype of patients with either a homozygous C282Y mutation or compound heterozygotes. 71 patients with a clinical diagnosis of hereditary haemochromatosis were identified through a questionnaire sent to physicians caring for hereditary haemochromatosis patients. Pertinent clinical data, in particular those reflecting iron body stores, were collected. Genotyping for the C282Y and H63D mutation of the HFE gene was performed. In 90% of the cases a mutation of the HFE gene was found. 86% of the patients were homozygous for the C282Y mutation, 4% were compound heterozygotes for the C282Y and the H63D mutation. Patients with the homozygous C282Y mutation showed a broad phenotypic spectrum that could not be accounted for by age or sex differences only. Our results demonstrate that within the Swiss population genetic testing can also identify the vast majority of patients with hereditary haemochromatosis. However, the diagnosis is not ruled out by a negative genetic test. Furthermore, a broad phenotypic spectrum is associated with the homozygous C282Y mutation in Swiss hereditary haemochromatosis patients. The implications of these findings for planning of widespread genetic screening for hereditary haemochromatosis in the general population are discussed. PMID- 11008305 TI - [Primary bilateral adrenal lymphoma]. AB - When abdominal imaging reveals the existence of unsuspected adrenal masses, a diagnostic strategy is necessary. We report the case of a woman presenting with pulmonary embolism, in whom abdominal ultrasound revealed voluminous masses in both adrenals without clinical or biological signs of hormone hypersecretion, but with mild primary adrenal failure. From a CT scan-directed needle biopsy of the right adrenal mass and subsequent staging we were able to diagnose a primary bilateral adrenal lymphoma, diffuse large B-cell type (REAL/WHO). On CHOP chemotherapy both adrenal masses decreased dramatically and the patient is in remission 18 months later. Primary adrenal lymphoma is a rare condition, since 65 cases have been reported to date. Histological diagnosis is nevertheless important, in view of the excellent response to specific therapy observed in some cases. PMID- 11008306 TI - [Unmasking an atrial flutter]. PMID- 11008307 TI - [Expert Health Promotion Panel of the Swiss Society of Prevention and Public Health. Total health promotion policy]. PMID- 11008308 TI - [Health and human rights--development of a questionnaire for measuring perceived human rights status]. AB - Health and human rights as interlinked concepts are a promising new avenue in public health. The aim of this study was the development of a reliable and valid instrument for the measurement of the human right status. The reliability and validity of this new questionnaire for human rights (HRS = human right scale) was measured within a group of Public Health students (n = 42) and the participants of the KORA-fracture-study. Test-retest-reliability was 0.73 (ICC: 0.69) and Cronbach's alpha was 0.66-0.79. The evaluation of the validity with the help of a factor analysis and a reconstruction study pointed out, that the perceived human rights status has to be treated as an interlinked and not divisible concept. Both study groups showed deficits for items in the dimension justice and participation. PMID- 11008309 TI - [Prevalence of smoking habits of Upper Austria students of the 7th and 8th grade and effect of smoking habits of family and peers]. AB - The aim of the study was to explore the prevalence of different smoking habits in a population of Austrian pupils, 12 to 15 years old, and the relationship of familial and peer group smoking customs with these habits. In 1997 a population based survey (International Study of Asthma and Allergies in Childhood, ISAAC) was conducted of all 7th and 8th grade school children of a district of Upper Austria. Information on the smoking habits of the adolescents, the family members, and of the peer as well as smoking habits of the teacher, gender, and age of the children was collected. The overall-prevalence of having ever smoked in this population is 57.8%. The percentage of eversmokers among the 12-year-olds is 50%. This amount increases to 63.8% among the 14- to 15-year-olds. The odds ratios for smoking daily is highest among those whose best friend smokes (OR: 70.63, CI: 9.19, 542.40). The risk of daily smoking increases also if the siblings of the juvenile (OR: 4.71, CI: 1.15, 19.35) or the mother (OR: 4.95, CI: 1.67, 14.70) smoke. If the father smokes the risk to smoke monthly is increased (OR: 2.09, CI: 1.28, 3.40). These results point to the fact that smoking prevention programes should take into account the influence of peers and family of the adolescents. PMID- 11008310 TI - A survey of institutional influenza vaccination in Switzerland. AB - In June 1998, a questionnaire was sent to evaluate the influenza vaccination practices in Switzerland: 429 health care institutions were to assess the level of influenza vaccination and the coverage of specific groups; each institution was required to specify whether vaccination coverage was known precisely (based on recorded data) or estimated. The response rate was 42.4%. Among institutions which responded, the mean accurate vaccination coverage rate for all patients was 40% and the estimated rate was 29%; these rates were slightly higher for people older than 65 years. For the entire staff, the accurate vaccination rate was 16% (14% estimated) whilst for the medical staff, the mean coverage was higher at 30% (measured) and 16% (estimated). The mean vaccination rate for patients and residents was 59% in the French speaking region of Switzerland, 54% in the Italian speaking canton, but only 37% in the German speaking region. The same ranking was found for vaccination coverage of medical staff: 21% in the French speaking region, 15% in the Italian speaking canton, and 13% in German speaking areas. These results suggest that cultural differences could play an important role in the attitudes and behaviour of the population regarding influenza vaccination programmes among the linguistic regions in Switzerland. PMID- 11008311 TI - [Congress report. 11. Clinical conference. Ultrasonography 2000-- trials, advances and trends]. PMID- 11008312 TI - [Boredom and diagnostic dilemma of splenic ultrasonography]. PMID- 11008313 TI - [Frequency, pattern and differential diagnosis of echogenic splenic changes: sonographic follow-up study]. AB - AIM: This study analyzes frequency, sonographic pattern and differential diagnosis of echogenic splenic foci and the value of ultrasound follow-up examinations. METHOD: Clinical data and ultrasound examinations of 137 patients with echogenic splenic foci were evaluated. Patients with uncertain diagnosis were identified and underwent a second examination (n = 31). RESULTS: Echogenic splenic foci are extremely rare. We found 137 cases in 12 1/2 years (12.2 of 10,000 ultrasound examinations). 5 sonographic patterns with differing but characteristic differential diagnosis were seen (round echogenic, round echogenic with halo and/or central liquefaction, round predominantly echogenic with hypoechoic parts, echogenic wedge-form, calcifications or gas). In patients with underlying malignant disorders the splenic foci were benign in 26 cases (34.7%), malignant in 36 cases (48.0%), and in 13 cases (17.3%) the diagnosis was uncertain. In patients with benign disorders the foci themselves were benign in 59 cases (95%), and in 3 cases (5%) the diagnosis was uncertain. CONCLUSION: Echogenic splenic foci are rare. Distinguishing between benign and malignant foci based upon sonographic patterns alone is only possible in exceptional cases. Short term follow-up ultrasound examinations taking into consideration the patients' underlying disease lead to the correct diagnosis in most cases. PMID- 11008314 TI - [Results of doppler sonography in normally functioning transjugular portosystemic shunts]. AB - PURPOSE: The objective of this study was to evaluate blood flow changes inside normally functioning transjugular intrahepatic portosystemic shunts (TIPS), using Colour Doppler ultrasound (CDUS) and pulsed Doppler ultrasound (PDUS). MATERIAL AND METHODS: In a prospective study 72 patients (mean age 54, range 36-78 years) underwent TIPS placement, portal angiography, CDUS and PDUS examinations. Along the TIPS Doppler measurements were taken at the portal in the central part and at the venous end of the TIPS. RESULTS: In well functioning TIPS the mean peak velocity (PV) on the portal side was 39 cm/s (range 22-72 cm/s), in the area of the incoming right portal end, branch 62 cm/s (range 40-109 cm/s) and at the site of the incoming hepatic vein 139 cm/s (range 88-220 cm/s). In the punctured portal and hepatic vein the mean PV was 25 cm/s (range 15-30 cm/s) and 18 cm/s (15-22 cm/s) respectively. CONCLUSION: A velocity gradient between the portal and the venous side of the TIPS is a normal finding, caused by the branches of the portal and hepatic vein, joining the TIPS from the side and is characteristic of a normally functioning TIPS. PMID- 11008315 TI - Comparison of portal venous flow in cirrhotic patients with and without paraumbilical vein patency using duplex-sonography. AB - AIM: In a prospective study we examined the effect of paraumbilical vein patency on the portal venous blood flow in patients with liver cirrhosis by Duplex sonography. In this context we investigated the influence of the aetiology and the severity of cirrhosis on the development of venous paraumbilical shunts. METHOD: In 70 patients (Child A: 16; Child B: 27; Child C: 27) with cirrhosis of different aetiologies the portal venous flow velocity, the blood flow volume, and the diameter of the portal vein were examined by Duplex sonography after an overnight fast. RESULTS: 16 patients had a patent paraumbilical vein. The mean portal venous flow velocity (19.2 +/- 7.8 cm/s) and the flow volume (1.29 +/- 0.50 l/min) were significantly higher in patients with paraumbilical vein patency than in patients without paraumbilical shunts (14.4 +/- 4.6 cm/s; p = 0.029 and 0.88 +/- 0.34 l/min; p = 0.007 respectively). The prevalence of shunts of the paraumbilical vein in cirrhotics was significantly lower in category Child A (6.3%) than in Child B (25.9%; p = 0.011) and Child C (33.3%; p = 0.006). CONCLUSION: The occurrence of relatively high portal blood flow velocities in cirrhotics must remind the examiner of the potential existence of spontaneous portosystemic shunts. Those shunts must be carefully looked for with colour Doppler ultrasound. In order to assess the true parenchymal blood flow we suggest to perform additional measurements of the right portal trunk and the left portal vein distal to the shunt. With an increase in the severity of liver cirrhosis the incidence of paraumbilical vein patency rises. PMID- 11008316 TI - [Hypertrophic pyloric stenosis: sonographic monitoring of conservative therapy with intravenous atropine sulfate]. AB - AIM: Ultrasound is the method of choice for the diagnosis of hypertrophic pyloric stenosis (HPS). The purpose of this study was to evaluate the usefulness of sonography in monitoring the efficacy of conservative therapy of HPS with intravenous atropine sulfate. METHOD: 21 infants with HPS under i.v. treatment with atropine sulfate were included. Pyloric sonomorphology, channel width and passage of gastric contents through the pyloric channel were monitored daily. The latter was examined with colour Doppler, too. If no clinical improvement was observed after 6-8 days, sonography and colour Doppler sonography played a crucial role in the decision whether to continue the conservative therapy or to perform pyloromyotomy. RESULTS: Conservative treatment was effective in 13/21 infants. In 8 patients therapy was continued as sonography demonstrated the passage of gastric contents despite lack of clinical improvement. In 4 patients, due to the sonographic findings, impending surgery could be cancelled. Colour Doppler sonography proved to be extremely useful in demonstrating passage of liquid through the narrowed pyloric channel. No significant change in pyloric morphology was seen. CONCLUSION: In HPS a conservative therapeutic approach with atropine sulfate is justified considering a success rate of 62% (13/21). During sonographic monitoring the detection of the passing of gastric content may be crucial for continuation and success of conservative therapy. In those cases colour Doppler sonography is a very useful method. PMID- 11008317 TI - [Imaging of intra-arterial hepatic port catheter systems by power-doppler sonography using contrast media--preliminary results]. AB - AIM: This study was designed to determine whether intra-arterial hepatic port catheter systems can be adequately detected by contrast-enhanced power-Doppler sonography. METHOD: 15 patients with a liver port system were investigated before chemotherapy. Examinations were performed with short bolus injections of the contrast medium Levovist in addition to angiographic imaging. RESULTS: Liver port systems were easily detected by the contrast-enhanced power-Doppler method. In 11 out of 15 patients a correct flow of the contrast medium via the port system was seen with both examination methods. One partially occluded hepatic artery was not identified by power-Doppler despite correct flow of the contrast fluid. In one of three patients showing an incorrect flow of the contrast medium the blood circulated primarily through the splenic artery due to a dislocation of the catheter tip. Circulation through both the hepatic and the splenic arteries was shown in a second patient and an occluded right hepatic artery was demonstrated in a third. All these findings were observed with both examination techniques. CONCLUSION: Power-Doppler sonography provides a reliable image of the port catheter system. This method can be used as a follow-up procedure to determine the state of arterial hepatic circulation during chemotherapy. PMID- 11008318 TI - [Three-dimensional ultrasound evaluation of the rotator cuff. Preliminary results of clinical application]. AB - AIM: Accurate sonographic diagnosis of rotator cuff lesions mainly depends on image quality and on the experience of the examiner. The present study was performed to determine whether the use of three dimensional (3D) sonography further increases the diagnostic yield of ultrasound examinations. METHOD: In a prospective study 25 patients who later underwent shoulder surgery of the rotator cuff were examined pre-operatively by ultrasound with two and three dimensional ultrasound. RESULTS: With three dimensional ultrasound rotator cuff lesions were more often correctly diagnosed (sensitivity of 100%) than with conventional 2D sonography (sensitivity of 91.7%). Specificity with both methods was 84.6% predictive value 92% and 88% respectively. In partial thickness tears, false negative results were twice as common with 2D imaging. CONCLUSION: Although our results are preliminary and have to be verified in a larger study, three dimensional ultrasound appears to facilitate diagnosing of rotator cuff lesions and partial thickness tears, in particular. PMID- 11008319 TI - [Biliary-digestive fistula with gallstone ileus--a sonographic diagnosis]. AB - We demonstrate a patient with a fistula between the gallbladder and the small intestine combined with a gallstone ileus of the small bowel, diagnosed by ultrasound. Plain abdominal X-ray only revealed small bowel obstruction. All the typical diagnostic criteria of a biliodigestive fistula could be detected by ultrasound. The patient was operated and discharged a few days later. We demonstrate that biliodigestive fistulas can be diagnosed by ultrasound. PMID- 11008320 TI - [On the fate of lost gallstones]. AB - The spilling of stones during laparoscopic cholecystectomy occurs in 10% of the operations. Late complications associated with this type of surgical procedure are abscesses and fistulas in the abdominal cavity and abdominal wall. The time elapsing between laparoscopy and clinical manifestation may be as long as 12 months. We present a case of a patient with localised abdominal pain in the upper right abdomen two months after laparoscopic cholecystectomy. Sonography and CT revealed a fat necrosis posterior of the rectus muscle. Examination of the histological specimen confirmed a fat necrosis caused by two small pigmented stones, spilled during laparoscopic cholecystectomy. PMID- 11008321 TI - From food constituent to dietary supplement and drug. Is there a role for antioxidants in treatment strategies? PMID- 11008322 TI - Therapeutic potential of glutathione. AB - Reactive oxygen species, formed in various biochemical reactions, are normally scavenged by antioxidants. Glutathione in its reduced form (GSH) is the most powerful intracellular antioxidant, and the ratio of reduced to oxidised glutathione (GSH:GSSG) serves as a representative marker of the antioxidative capacity of the cell. Several clinical conditions are associated with reduced GSH levels which as a consequence can result in a lowered cellular redox potential. GSH and the redox potential of the cell are components of the cell signaling system influencing the translocation of the transcription factor NF kappa B which regulates the synthesis of cytokines and adhesion molecules. Therefore, one possibility to protect cells from damage caused by reactive oxygen species is to restore the intracellular glutathione levels. Cellular GSH concentration can be influenced by exogenous administration of GSH (as intravenous infusion or as aerosol), of glutathione esters or of GSH precursors such as glutamine or cysteine (in form of N-acetyl-L-cysteine, alpha-lipoic acid). The modulation of GSH metabolism might present a useful adjuvant therapy in many pathologies such as intoxication, diabetes, uremia, sepsis, inflammatory lung processes, coronary disease, cancer and immunodeficiency states. PMID- 11008323 TI - Glutathione in the prevention of cisplatin induced toxicities. A prospectively randomized pilot trial in patients with head and neck cancer and non small cell lung cancer. AB - PURPOSE: Glutathione has been shown to be an effective chemoprotector against cisplatin-induced side effects in patients with ovarian cancer. In view of this fact, we performed a randomized clinical pilot-trial in the management of other solid tumors in order to compare application of Glutathione to intensive hydration in patients undergoing chemotherapy with a regimen including cisplatin. PATIENTS AND METHODS: Twenty patients suffering from advanced non small cell lung cancer (n = 6) or head- and neck cancer (n = 14) were enrolled in the study. All patients received 80 mg/m2 cisplatin along with etoposide or 5-fluorouracil every 4 weeks. Patients randomized to application of Glutathione (n = 11) received 5 g of Glutathione immediately before application of cisplatin followed by 2000 ml of normal saline. Patients in the control group (n = 9) received 2000 ml electrolyte infusion before and 2000 ml of normal saline with forced diuresis after cisplatin. RESULTS: The intensity of hematologic toxicity was significantly less pronounced in patients treated with Glutathione than in the control group (hemoglobin: 10.7 vs 9.5 mg% respectively, p = 0.039; white blood cell count 3.3 vs 2.2 x 103/microliter respectively, p = 0.004; platelets 167 vs 95 x 103/microliter respectively, p = 0.02), whereas in terms of non-hematologic toxicity no difference was observed. Objective remission occurred in 6 out of 11 evaluable patients from the group receiving Glutathione (55%; complete remission: 9%; partial remission: 46%), and in 4 out of 8 evaluable patients from the control group (partial remission: 50%). However, there was no statistical difference in terms of response and overall survival (13.5 months vs. 10.5 months) between the two groups. CONCLUSIONS: Application of Cisplatin and Glutathione seems to be safe and feasible and the antitumoral efficacy of cisplatin is apparently not impaired by the concomitant use of Glutathione in patients with solid tumors. PMID- 11008324 TI - Efficacy of standard treatment in patients with Hodgkin's disease--a single center experience. AB - Sixty-two consecutive patients with newly diagnosed Hodgkin's Disease (HD) were treated at our institution between 1991 and 1998 in accordance with protocols which we considered as optimal standard therapy. In patients with localised disease, extended field irradiation alone was used initially but short duration chemotherapy followed by involved field irradiation was given later. Initially, MOPP/ABV chemotherapy was considered the standard treatment regimen for advanced HD, subsequently it was replaced by ABVD treatment. At 5 years patients with localised HD have a projected overall survival of 95% and a failure-free survival of 72%. Relapses in patients treated with radiotherapy alone (6/16 cases) could be successfully managed with chemotherapy in most cases (5/6). In advanced HD, projected 5-year survival and failure-free survival are 100% and 80%, respectively. No treatment-related deaths or secondary malignancies were registered. PMID- 11008325 TI - [Hormone replacement therapy with a transdermal estradiol gel and oral micronized progesterone. Effect on menopausal symptoms and lipid metabolism]. AB - In a multicenter observational study, the efficacy and acceptance of two different regimens of postmenopausal hormone replacement therapy in the form of a combination of 17 beta-estradiol in percutaneous gel application and micronized oral progesterone were evaluated. Forty-eight patients (aged 40-66 years) received 2.5 g estradiol gel plus either continuously micronized progesterone 100 per day (group A) or, sequentially, 200 mg per day between day 16 and 25 of a monthly cycle (group B) for two months. A significant reduction in typical menopausal symptoms, especially vasomotor complaints like hot flushes or sweating, was observed in both groups (score average at the beginning for hot flushes: 2.0 in group A and 1.8 in group B; after two months of treatment, 0.7 in group A and 0.4 in group B). Cholesterol levels were slightly reduced but statistically significant (235.9 +/- 49.55 mg/dl vs. 226.3 +/- 52.24 mg/dl; p < 0.05) only in group A; a trend towards lower cholesterol was observed in group B (236.5 +/- 47.82 mg/dl vs. 227.4 +/- 44.72 mg/dl). Lipoprotein (a) was also significantly reduced in group A (32.57 +/- 36.52 mg/dl vs. 28.28 +/- 31.03 mg/dl in group A; 31.7 +/- 28.42 mg/dl vs. 28.34 +/- 23.71 in group B; p < 0.05). The overall acceptance of this therapy was excellent or good in 91.3% of group A and 92.8% of group B patients. PMID- 11008326 TI - L-Tryptophan does not influence acute ventilatory response at moderate altitude. AB - Altitude induced insomnia is a very common symptom in mountaineering. Conventional hypnotics such as benzodiazepines potentially can be associated with untoward side effects because they can impair ventilatory adaptation to hypoxia at altitude. The objective of our study was to evaluate the effect of a alternative potentially sedative drug, L-tryptophan on ventilation at moderate altitude. STUDY DESIGN, METHODS AND RELEVANT RESULTS: Randomised, double blind, placebo controlled crossover trial. Blood gas analysis of 8 healthy subjects was performed before and one hour after oral administration of 500 mg L-tryptophan or placebo at altitudes of 171 m and at 3,000 m. PaO2 and PaCO2 before and after L Tryptophan or placebo medication did not change significantly at neither level of altitude investigated. CONCLUSION: L-tryptophan does not impair ventilatory adaptation to mild hypoxia at moderate altitude. PMID- 11008327 TI - Successful use of continuous intravenous prostacyclin in a patient with severe portopulmonary hypertension. AB - INTRODUCTION: Portopulmonary hypertension, defined by a mean pulmonary artery pressure > 25 mm Hg in the presence of normal pulmonary capillary wedge pressure and portal hypertension, is a known complication of end-stage liver disease that has been associated with high morbidity and mortality at the time of liver transplantation. DESIGN: Descriptive case report. PATIENT: A 32 year old male patient suffering from end-stage hepatitis C liver cirrhosis presented with severe portopulmonary hypertension. At presentation the following pulmonary hemodynamics were measured: systolic pulmonary artery pressure (PAP) 76 mm Hg, mean PAP 42 mm Hg, pulmonary vascular resistance index (PVRI) 931, pulmonary capillary wedge pressure (PCWP) 9 mm Hg, and cardiac output (CO) 4.03 l/min. INTERVENTION: After acute hemodynamic testing the patient received 8 ng/kg/min epoprostenol (prostacyclin) by continuous intravenous infusion with an infusion pump. Hemodynamic evaluation was performed monthly by transthoracic echocardiography and right heart catheterisation after 5 months. RESULTS: After 5 months of continuous therapy right heart catheterisation revealed the following hemodynamics: systolic pulmonary artery pressure (PAP) 59 mm Hg, mean PAP 32 mm Hg, pulmonary vascular resistance index (PVRI) 561, pulmonary capillary wedge pressure (PCWP) 7 mm Hg, and cardiac output (CO) 6.95 l/min. This presents a decrease in systolic pulmonary artery pressure of approximately 22%, a decrease in mean pulmonary artery pressure of approximately 30%, a decrease in pulmonary vascular resistance of approximately 40% and an increase in cardiac output of approximately 73%. Echocardiography demonstrated a decrease in estimated systolic pulmonary artery pressure of about 37% after 8 months of therapy. No complications were observed during epoprostenol therapy. CONCLUSION: In this adult patient suffering from end-stage liver disease and portopulmonary hypertension, administration of continuous intravenous epoprostenol resulted in significant reduction of pulmonary hypertension and therefore in acceptance for orthotopic liver transplantation. Utilisation of this new therapeutic strategy might be a helpful pharmacological tool for patients with portopulmonary hypertension to make them acceptable for orthotopic liver transplantation. PMID- 11008328 TI - [Hitler and the anti-smoking movement in the time of national socialism]. PMID- 11008329 TI - [Effect of ACE-inhibitors on left ventricular remodeling after acute anterior or posterior myocardial infarct. Cine- magnetic resonance tomography study]. AB - The question whether patients with posterior infarctions (PMI) have a comparable benefit of an ACE-inhibitor therapy to those with anterior infarction (AMI) is still open. The study was undertaken to investigate the different influence of ACE inhibitors on the remodeling of the left ventricle after AMI or PMI. 52 patients (Pt.) (17 female, 38-73 years) were randomized to receive either 25-75 mg/day captopril (C) or 5-20 mg/day fosinopril (F) beginning on day 7 after acute myocardial infarction. 28 Pt. had AMI, 24 Pt. PMI. Infarct size was determined by the creatine kinase integral method. 50 Pt. were examined by cine magnetic resonance imaging 1 and 26 weeks after infarction. We determined: left ventricular end-diastolic (LVEDVI) and end-systolic (LVESVI) volume index, ejection fraction (EF), infarction weight (IW), left ventricular muscle mass (MM), systolic wall thickening (SWT) and motility (MOT) of the vital myocardium, and clinical behavior according to the guidelines of the New York Heart Association (NYHA). The results were compared with those of a sample (V) without ACE inhibitor therapy (10 females, 21 males, 36-75 years, 19 AMI, 12 PMI). There were no significant differences between C and F. Without ACE-inhibition therapy LVEDVI increased by 28.2% in AMI, by 18.4% in PMI (p < 0.001), with ACE inhibition by 13.7% in AMI and by 9.9% in PMI (p < 0.001). LVESVI increased in V by 40.1% in AMI, by 28.5% in PMI (p < 0.001). With ACE-inhibitor we found an increase of 11.2% in AMI and 5.3% in PMI (p < 0.001). EF decreased without ACE inhibitor by 18.7% in AMI and by 10.2% in PMI (p < 0.001), with ACE-inhibition increased by 4.3% in AMI and PMI, respectively (n. s.). NYHA got better in all groups, by 17.4% in AMI and 20.8% in PMI without ACE-inhibitor (n.s.), by 45.5% in AMI and 31.6% in PMI with ACE-inhibitor (p < 0.001). IG increased by 15.5% in AMI and 8.8% in PMI in V (p < 0.001), by 11.2% in AMI and 5.3% in PMI with C or F (p < 0.001). MM got bigger in V by 16.6% in AMI and 12.7% in PMI (p < 0.05), with ACE-inhibitor by 11.7% in AMI and 8.0% in PMI (p < 0.05). sWD increased by 12.9% in AMI and by 6.7% in PMI in V (p < 0.01), by 37.1% in AMI and 88.0% in PMI with C or F (p < 0.001). MOT decreased by 39.6% in AMI and 14.9% in PMI without ACE inhibition (p < 0.001) and increased by 4.3% in AMI and by 5.0% in PMI with ACE inhibitor (n. s.). All differences between V and the ACE-inhibitor groups were significant. Even patients with PMI clearly benefit from ACE-inhibitor therapy, but less than those with AMI. Captopril and fosinopril show no different effects after myocardial infarction. PMID- 11008330 TI - [Certain aspects of plastic surgery in the head and neck area]. AB - The spectrum of plastic surgery in the head and neck area is wide. The indications for as well as potentials and limits too, aesthetic surgery cannot be pressed into a rigid framework. The operative strategy will substantively depend on several factors, including objectivated local findings from, as well as general situation and motivation, of the patient. In plastic surgery the psychological situation of the patient is very often paramount, especially as regards cosmetic aspects. Sometimes the declared motives of the patient hide emotional disorders. Several examples show the practicability and limits. The possibilities of corrective action depend not only on the local findings but also on the surgeon's experience and skill. PMID- 11008331 TI - [Osteoporosis in Turner syndrome with chromosomal mosaicism (45,XO/46,XY). A case report]. AB - We report a rare case of Turner's syndrome (UTS) due to a gonosomal mosaic 45,X0/46,XY with the main clinical feature of several fractures in the course of osteoporosis. The bone mineral density (BMD) of the lumbar spine and the hip measured by DXA showed osteoporosis. The other clinical investigations including laboratory parameters presented beside an estrogen deficiency due to primary amenorrhea, a small thyroid with hypothyroidism, increased renal calciuria and increased markers of bone metabolism. Beside the supplementation of estrogen and thyroid hormone deficits, only a combination of different bone-associated drugs could normalize the bone mineral density and the bone turnover. During a two years' follow-up period no further fractures occurred. Furthermore, theories regarding the occurrence of osteoporosis and other complex pathologies in UTS are discussed with the conclusion that osteopenia in UTS is probably based on a multifactorial genesis. Due to these complex pathologies during adulthood, patients with UTS should get multidisciplinary care in order to reduce morbidity and preserve quality of life. PMID- 11008332 TI - [History of neurosurgical treatment of gunshot wounds]. AB - The provision of neurosurgical managing care in the German field hospital during the IFOR- and SFOR-mission in the former Yugoslavia is a novum within the NATO. Penetrating head trauma is by far the most common type of central nervous system trauma observed in a military combat setting. There is a distinct advantage realized by early intervention following penetrating head trauma with increased intracranial pressure. As well as the experience of Ernst von Bergmann a hundred years ago during the Prussian-Austrian-War as the reports of the operation Desert Storm confirm the advantage of an early neurosurgical intervention of gunshot wounds of the head. The decision of the German Armed Forces to provide forward located neurosurgical maintenance is justified and will lead to major reduction of time between injury and surgery. In the future neurosurgical managing care in combat settings will be supplemented and improved by peer-to-peer networks through the World Wide Web. PMID- 11008333 TI - [Cesarean section by patient request--permissive obstetrics or acceptable as self determined delivery?]. PMID- 11008334 TI - [Nausea and vomiting in early pregnancy/hyperemesis gravidarum. Current status of psychosomatic factors]. AB - BACKGROUND: Although Hyperemesis gravidarum (HG) is still a disease of unknown aetiology psychosocial aspects are involved in the genesis of this disease. MATERIALS AND METHODS: Based on experiences in our department and on actual literature psychosocial aspects and therapeutic approaches of HG are presented. RESULTS: Predisposing factors of HG are stress, inadequate information about pregnancy or delivery and problems in the marital relationship. Feelings of ambivalence towards pregnancy, a distorted perception of reality, the unconscious refusing of a wanted child and the worry about a relation between nutrition of the foetus and the HG have influence on the development of the HG. The role of the social context, especially the mother, of expectations towards vomiting and of secondary gain in the genesis and the evaluation of HG is explained. Therapeutical approaches beside the known somatic therapies are acupuncture, the electric stimulation of the N. vestibularis and different methods of psychotherapy, which can include the partner or other persons from the social context. In some cases an intensified therapy under in-patient conditions can improve therapeutic results. DISCUSSION: Psychosocial factors are involved not only in the genesis of HG, but also in the duration and intensity of symptoms as well as in the resistance especially, towards isolated somatic therapeutical approaches. CONCLUSIONS: When psychosocial factors are taken into account in the search for a cause and the treatment of HG results of treatment are more successful and persist over a longer time. PMID- 11008335 TI - [Doppler ultrasound examination of fetal umbilical arteries of the intra abdominal segment in normal singleton pregnancies]. AB - BACKGROUND: In the management of high risk pregnancies, blood flow measurements in the umbilical arteries (Aa. umb) have reduced perinatal morbidity and mortality. No consensus has been reached about the optimal location of measurements along the umbilical cord. The present study analyses the blood flow in both umbilical arteries where they pass by the fetal bladder. PATIENTS AND METHODS: In 48 normal pregnancies, Doppler examinations were used to measure the pulsatility indices (PI). From the 20th week of pregnancy onward, the PI were measured once every four weeks in the intraabdominal segments of the right and the left Aa. umb. lateral to the fetal bladder. Measurements were also done in a free loop and at the placental insertion. RESULTS: For all locations, normal ranges were calculated. The PI at all four sites decreased as the pregnancies were approaching term. In the III. trimester, the standard deviations of PI were smaller than in the II. trimester. The PI at the intraabdominal segment of the right and the left umbilical arteries turned out to be similar to each other and exceeded the values measured at the free loop and at the placental insertion respectively. CONCLUSION: The intraabdominal segments of the umbilical arteries were defined as a new location for PI measurements. When adopting a fixed place of measurement, it is likely that the reproducibility of PI measurements will be increased. DISCUSSION: The higher PI at the intraabdominal segment of the umbilical arteries are caused by the more central location compared to the placental insertion. PMID- 11008336 TI - [Intercellular adhesion molecular 1, a diagnostic serum marker in chorioamnionitis, pre-eclampsia and HELLP syndrome]. AB - INTRODUCTION: Chorioamnionitis and pregnancy-induced hypertension both are extremely feared complications of human pregnancy. Activation or disturbance of normal endothelial cell function may be involved in the pathogenesis of both kinds of disease. The aim of our study was to compare the diagnostic value of soluble intercellular-adhesion-molecule-1 (ICAM-1) with that of C-reactive protein (CRP) and white blood cell count for the detection of chorioamnionitis in patients with preterm labor. In addition, we examined if concentrations of ICAM-1 were also increased in case of preeclampsia or HELLP-syndrome. MATERIALS AND METHODS: ICAM-1, CRP and leucocyte count were estimated in 50 cases of normal term delivery, 97 cases of uncontrollable preterm labor, 16 cases of preeclampsia and 9 cases of HELLP-syndrome before delivery. RESULTS: From 97 women delivering preterm, chorioamnionitis was histologically confirmed for 48 women. Maternal serum levels of ICAM-1 (p < 0.001), CRP (p < 0.001) and leucocyte count (p < 0.02) were significantly higher in the group of preterm delivering patients (< 37 weeks gestation) with histologically confirmed chorioamnionitis in comparison to preterm delivering patients in the absence of chorioamnionitis. In the group of patients delivering preterm (< 37 weeks gestation) because of preeclampsia or HELLP-syndrome, ICAM-1 (p < 0.001), as well as CRP (p < 0.006) concentrations were also significantly increased in comparison to patients delivering preterm in the absence of chorioamnionitis. CONCLUSIONS: Elevated levels of ICAM-1 in the serum of pregnant women may be considered as an important risk factor for the development of complications in pregnancy associated with inflammatory induced changes of maternal endothelial cell functions. PMID- 11008337 TI - [Significance of uterine contractions for adaptation of the newborn infant before cesarean section]. AB - There are quite a few publications on the influence of labour on the adaptation of neonates after elective cesarean sections. Many authors recommend the induction of labor prior to the cesarean section. They found that this improves the adaptation of the infants. MATERIAL AND METHOD: All cesarean sections between 1991 and 1996 are recorded. Mature neonates (> 36 weeks of pregnancy) were evaluated separately from premature infants (< 36 weeks of pregnancy). As target parameters we chose 1. the condition of the neonates (characterized by the Apgar Score, the pH of the umbilical cord artery, and the requirement of artificial respiration). 2. the necessity of transfer to the neonatologic intensive care unit. Concerning these parameters, the immature neonates (> 36 weeks of pregnancy) were evaluated separately from the mature infants (< 36 weeks of pregnancy). In the latter group we finally evaluated the data of those who were not delivered by a cesarean section because of intrauterine fetal distress. RESULTS: The adaptation of the premature neonates was so closely related to the gestational age that all the other variables were of no statistically significant influence. Thus also the factor "labour" was of no influence. The results in the group of the mature infants were completely different. Between week 36 und 42 of pregnancy the gestational age was of no influence on the adaptation of the neonates. Statistically noticeable however was the influence of preoperative labour: The adaptation of infants delivered by cesarean-section after labour was remarkably worse than the adaptation of those who were delivered without labour prior to the operation. This statement is also correct when one rules out those infants who were delivered by a so called emergency cesarean section. When evaluating the group of neonates in which the cesarean section was not performed because of intrauterine fetal distress we could find no influence of preoperative labour on the adaptation of the infants. All other variables examined (e.g. maternal age, parity, duration of labour, method of anaesthesia, twin-pregnancy) were no factors of influence on the adaptation of the babies. CONCLUSION: From our results we cannot support the recommendation to induce labour prior to any elective cesarean section by infusion of oxytocin or to wait for the spontaneous onset of labour. There ist no reason to change our policy to perform an elective cesarean section as near as possible to term before the onset of labour. PMID- 11008338 TI - [Fetal complications after failed pregnancy termination in the first trimester]. AB - BACKGROUND: Two different methods are in use for induction of abortion: either dilation and curettage (D & C) or medical termination with antigestagens and/or prostaglandins. The risk of continuing pregnancy after D & C ranges from 0.023 to 1% and is inversely correlated with gestational age. METHODS AND RESULTS: Several authors reported fetal anomalies (limb and oro-facial defects) in such cases. For the medical induction of abortion a combination of Mifepristone and Misoprostole is the treatment of choice in order to reduce the risk of ongoing pregnancy. Using single drug therapy carries an increased risk of failure and of subsequent fetal anomaly. There seems to be an increased incidence of limb defects and Mobius' syndrome in fetuses born after failed termination of pregnancy, but normal fetal development is entirely possible. CONCLUSION/DISCUSSION: The patients concerned must be informed in detail of the risks and probable consequences which must be included in the counselling concept of the treating gynaecologist. PMID- 11008339 TI - [Malignant melanoma and pregnancy]. AB - BACKGROUND: Incidence of malignant melanoma has more than doubled in the past 10 years. Two thirds of malignant melanoma arise from melanocytes, the pigmented cells of epidermis and dermis, in one third of the cases from preexisting nevi. CASE REPORT: We report on a 39 year-old pregnant woman with liver metastases after surgical removal of a malignant melanoma two years before. The mother died in the course of progredient liver insufficiency, a male baby had been delivered with a gestational age of 26 weeks + 4 days via cesarean section 11 days before and developed normally. RESULTS: Early diagnosis and operative excision are decisive for survival, in advanced stages chemotherapy does not prolong survival. CONCLUSIONS: Interdisciplinary co-operation of the pediatrician, obstetrician, and oncologist is of utmost importance. DISCUSSION: Melanoma and pregnancy represent a difficult situation for the patient, her partner and the treating physicians. Here the wish to have a child meets the potentially deadly cancer disease. PMID- 11008340 TI - [Are guidelines needed for diagnostic psychological testing in child- and adolescent psychiatry?]. PMID- 11008341 TI - [Utilization of psychological tests for diagnosis in German child and adolescent psychiatry: results of a survey]. AB - 92 inpatient and outpatient child and adolescent psychiatric institutions all over Germany were surveyed on practice and attitude towards psychological tests. Leading psychologists were contacted by mail and given a 15 item questionnaire. All institutions reported keeping at least a small test-inventory and using psychological tests in diagnostics. Half of the clinics carried out individual tests obligatory in the routine. Additionally, one third applied self-constructed instruments, which in the minority of cases had been psychometrically investigated or published. Computer-aided testing was established in one third of the institutions. Colleagues, test-compendia, scientific journals and the internet are quite often consulted for test-information. Intelligence measures are the most frequently administered test in child and adolescent psychiatry, while structured interviews are applied rarely. Among the highly frequent used individual tests were the Wechsler-Intelligence-Scales for children and adults, the CFT-scales, the projective Sceno-test and the d2-attention endurance test. One forth of the surveyed clinics reported refusal of individual or groups of tests, including projective and cognitive assessments. Thereby, ethic aspects were never mentioned as a reason for rejection. A range of diagnosticians reported to miss adequate measures for diverse constructs, e.g. learning disabilities. Furthermore, new norms and up-dates for regularly used instruments were often demanded. Some single survey-results also give reason for critically reviewing aspects of test-usage in German child and adolescent psychiatry. PMID- 11008342 TI - [Psychological testing in theory and practice exemplified by intelligence testing]. AB - OBJECTIVES: The U.S.-American standards of quality for psychological diagnostics tests were published in 1998 in German translation under the title "Standards fur padagogisches und psychologisches Testen". The relevance of these Standards for studies of psychological testing in child and adolescent psychiatry is discussed. Important intelligence tests will be assessed on the basis of these standards for studies of psychological testing in children and adolescents. METHODS: Relevant standards were presented and discussed. The intelligence tests HAWIK-R, HAWIK III, K-ABC, AID, PSB, I-S-T 70, SPM, CFT 20, CMM-LB, SON-R 2 1/2-7, SON-R 5 1/2 17 were analyzed with regard to whether or not they fulfilled the Standards under inclusion of the test manuals, test reviews, and empirical studies from the literature. RESULTS: The intelligence tests listed differ widely in the extent to which they meet the Standards. CONCLUSIONS: The Standards appear, at least in part, to be relevant to studies of psychological testing in child and adolescent psychiatry. The intelligence tests used in such studies should be critically examined with regard to their quality and whether or not they are suitable for the issues to be resolved. Given their completely outdated standard, it is recommended to refrain from further use of PBS and I-S-T 70. PMID- 11008343 TI - [Neuropsychological testing in childhood and adolescence]. AB - Among a number of issues and disorder pictures neuropsychological diagnostics should occupy a solid position, both in the initial assessment, as well as in that of the course. With an eye towards its use in child and adolescent psychiatry, a descriptive definition of this area of psychological diagnostics must be provided. In so doing, several essentially different approaches arise, whose effects on psychometrics and interpretation must be examined. Regardless of the fact that the current availability of standardized neuropsychological diagnostics is satisfactory only in certain areas, the methods used in clinical practice and/or research are presented. In addition to the individual test batteries, individual test methods for the areas of visual and auditory memory functions, attention functions, functions associated with speech, and executive functions are discussed. The current status of neuropsychological diagnostics generates essential tasks towards its further development. This is true with regard to goals attainable in the short term, such as adaptation or standardization of available instruments, as well as for long-term research tasks. PMID- 11008344 TI - [Diagnostic testing methods for skill assessment in reading, writing, and arithmetic. A critical review]. AB - OBJECTIVES: The diagnosis of a specific developmental disorder of reading, writing and arithmetic can be made based upon individually applied standardized methods for testing scholastic achievement and IQ. To make the choice of suitable methods easier for the administrator of the test, a critical survey of German language methods for assessing skills in reading, writing and arithmetic is presented. METHODS: Test intention and psychometric properties for scholastic achievement are summarized. The methods are assessed with regard to their utility in the diagnosis of congenital alexia and dyscalculia. Supplementary suggestions for clinical assessment are given. RESULTS: In summary, beyond the primary school area there is a lack of current standardized methods that meet the current standards of quality. Particularly the assessment of arithmetic skills above those of the 4th grade level require resorting to methods of dubious curricular validity. Coverage is once again better for the upper elementary and middle school levels. CONCLUSIONS: There is a need for new constructions, respectively a need to update published scholastic achievement tests. PMID- 11008345 TI - [Gottinger Form Reproduction Test (GFT)--results from an unselected sample]. AB - The Gottingen Form Reproduction Test (GFT) is a relatively old method of assessing visuo-motor coordination that is still often performed in clinical practice. Based on a sample of 72 unselected second-graders, it was shown that the mean percent ranks are too high. GFT percent ranks then should be interpreted with extreme caution in the course of clinical diagnosis. PMID- 11008346 TI - [Are eight-year olds already suffering from panic disorder? Investigation of prevalence with focus on panic disorder and agoraphobia]. AB - A representative sample of 826 eight-year-old second-graders in Dresden was examined in a two-stage procedure to ascertain the 6-month prevalence of DSM-IV anxiety disorders. Particular care was taken to assess panic disorder. The eight year-olds had already been interviewed personally in a screening session and in 230 children an individual diagnostic examination with a structured diagnostic interview was carried out. The total prevalence of anxiety disorder was 9.5%. No case of either panic disorder or agoraphobia was found. For the diagnosis of agoraphobia the DSM-IV diagnostic criterion was lacking in which situations typical to agoraphobia are feared or avoided because escape would be difficult or it would be difficult to reach help. Eight-year-olds do not forge this cognitive link. In agoraphobic situations 2.5% of the children (almost exclusively, girls) suffer from a specific phobia. The prevalence of anxiety disorders is markedly higher among girls than in boys. PMID- 11008347 TI - [The autopsy in crisis]. AB - In the Western world the autopsy rate is declining at an alarming rate. In the United States of America the rate in some academic hospitals is less than 7% of all hospital deaths. This decline has been documented and deplored in many countries, articles and books. Suggestions on how to resuscitate the autopsy range from mandatory in all hospital deaths to economic bonuses to the doctors obtaining the highest autopsy rate. All in vain, the autopsy decline continues. Pathologists deploring this decline blamed clinical colleagues, new social attitudes, the litigious nature of modern society, but few have questioned a procedure little changed in more than a century. Perhaps the time has come to abandon the "classic" autopsy and rethink the procedure so as to make it useful, alluring and indispensable for the contemporary, concerned clinician. PMID- 11008348 TI - [Evaluation of acute effect of GnRH administration on leptin secretion in normal and hyperandrogenic women]. AB - BACKGROUND: Several studies suggest that leptin modulates the reproductive axis function. Leptin may stimulate release of GnRH from hypothalamus and of gonadotrophins from the pituitary. A synchronicity of LH and leptin pulses has been described in healthy women and in patients with polycystic ovarian syndrome (PCOS), suggesting a relationship between the episodic secretion of LH and leptin. In vitro experimental studies have demonstrated that leptin administration promotes GnRH-LH release. However it is not established whether GnRH promotes the episodic secretion of leptin. AIM: To assess the response of LH and leptin to the administration of a GnRH bolus in hyperandrogenic and healthy women. PATIENTS AND METHODS: Eleven hyperandrogenic and eleven healthy women of similar age and body mass index (BMI) were studied. Under basal conditions three blood samples were collected every 30 min before and after the administration of a GnRH bolus (100 micrograms). LH and leptin concentrations were measured in all samples. Testosterone, SHBG and estradiol were determined in the first sample. For data analysis, the increment of LH and leptin between 0-30 and 0-60 min was calculated. The LH and leptin areas under the curve (AUC) before and after GnRH administration were also calculated in both groups. RESULTS: After GnRH administration an increment in LH concentrations was observed in both groups; however, leptin concentrations were not modified. In both groups LH area under the curve increased after GnRH administration; however, the leptin area was not modified. CONCLUSIONS: These results suggest that circulating leptin concentration is not modulated by GnRH-LH. PMID- 11008349 TI - [Improvement in inspiratory muscle function in patients with mitral stenosis after percutaneous mitral valvuloplasty. Preliminary communication]. AB - BACKGROUND: It has been proved that there is an inspiratory muscle dysfunction in mitral stenosis; Although its causes still remain unknown. AIM: To evaluate the effect of percutaneous balloon mitral valvuloplasty (PMV) on inspiratory muscle performance (IMP) in patients with mitral stenosis (mitral area < 1.5 cm2). PATIENTS AND METHODS: We studied IMP in 8 patients (35 +/- 10 years) before and 3 months after successful PMV. Inspiratory muscle strength was studied by measuring maximal statistical inspiratory mouth pressure (MIP). Endurance was evaluated using a two minute incremental threshold loading test in order to obtain the maximal sustainable inspiratory pressure (SIP), with the maximal sustainable load (MSL) the patients could sustain for 2 minutes. RESULTS: Mitral valvuloplasty increased mean cardiac index from 3.1 +/- 0.3 to 4.15 +/- 0.3 l/min/m2 (p < 0.01), and significantly decreased mean pulmonary and capillary pressures. The MIP value(118 +/- 6 cmH2O), similar to that of normal group, increased to 137 +/- 7 cmH2O (p < 0.01). SIP and maximal sustainable load were 52 +/- 3 cmH2O and 294 +/- 29 g respectively, lower than normal subjects (p < 0.05). They increased after PMV to 80 +/- 3 cmH2O and 463 +/- 26 g respectively (p < 0.001). CONCLUSIONS: PMV improved inspiratory muscle function in patients with severe mitral stenosis, probably secondary to a decrease work of breath and improvement of ventricular function. PMID- 11008350 TI - [Antihypertensive effects of valsartan. National multicentric study]. AB - BACKGROUND: Valsartan is an angiotensin AT1 receptor antagonist. Clinical studies have shown that this is an effective and well tolerated drug. AIM: To compare valsartan with other commonly used antihypertensives in a National multicentric study. MATERIAL AND METHODS: Between 1997 and 1998, a pharmacological surveillance of patients with mild or moderate hypertension, coming from different regions of the country, was carried out. Patients were followed during 8 weeks. In the group treated with valsartan, the initial dose was 80 mg/day. After four weeks of therapy, and according to patient's response the dose of valsartan or of the other antihypertensives was modified. RESULTS: Five hundred ninety three patients, aged 59 years old as a mean, were included in the study. Of these 434 (265 female) received valsartan and 159 (101 female) other antihypertensive drugs. Fifteen percent of patients treated with valsartan and 25% of patients with other medications required a dose change at four weeks, due to lack of response (p < 0.01). At week eight, 1.5% of patients with valsartan and 17.4% of patients with other medications reported adverse effects such as cough, headache, edema or flushing (p < 0.01). CONCLUSIONS: In this Chilean observational multicentric study, valsartan proved to be more effective and to cause less adverse effects than commonly used medications in hypertensive patients. PMID- 11008351 TI - [Variability determination in chromatin condensation and seminal alpha glucosidase with relation with the spermiogram]. AB - BACKGROUND: Sperm functional tests as an addition to semen analysis have been used to study the fertilization ability of spermatozoa. Besides the usual variability of the seminal analysis an individual variability in the results of functional tests has been recently found. AIM: To evaluate in a three months period, the individual variability of sperm parameters and sperm maturation using the chromatin condensation test and epidydime a-glucosidase (that allows to discriminate obstructive processes). MATERIAL AND METHOD: The evaluation was carried out in two donors (12 samples) apparently in good health. One of them presented evident semen analysis alterations (donor 1) and the other was considered normal under the WHO standards (donor 2). RESULTS: The averages for donor 1 were: Sperm count 24 x 106 sperm/ml (range 10-58 x 106 sperm/ml), morphology 31.8% (range 30-35%), total motility 33% (range 20-42%), sperm maturation 38% (range 28-78%), a-glucosidase 8.65 (U/ml (range 5-10 (U/ml). The averages for donor 2 were: Sperm count 96 x 106 sperm/ml (range 50-140 x 106 sperm/ml), morphology 32.2% (range 30-35%), total motility 69% (range 58-78%), sperm maturation 17% (range 7-30%), a-glucosidase 36.9 (U/ml (range 20-82 (U/ml). CONCLUSIONS: These results show that significant variations can be found in the sperm parameters and in seminal plasma a-glucosidase; however these variations are generally maintained at the normal or abnormal ranges for each individual, except the sperm morphology that was constant and with low variation in both donors. The determination of the chromatin condensation in the semen analysis gives an additional information about the grade of sperm maturation and would be of great value for differentiating between sperm samples that show similar morphology values. PMID- 11008352 TI - [Clinical and molecular characterization of autosomal recessive chronic granulomatous disease caused by p47-phox deficiency]. AB - BACKGROUND: The cytosolic protein p47-phox (phagocyte oxidase) is one of the essential components of the superoxide generating system in phagocytes and its defect causes approximately 30% of the chronic granulomatous disease (CGD) cases. AIM: Two patients were studied, belonging to the same family, without a consanguinous background, in which deficiency or absence of superoxide generation was found together with recurrent and severe infections in one case and benign infections in the second. METHODS: The presence of gp91-, p67- and p47-phox in patients and controls was determined by Western Blot analysis of granulocytes. Sequencing of PCR amplified DNA was performed by an enzymatic method. RESULTS: Western Blot analysis showed normal expression of gp91 and p67 and absence of p47 phox. The molecular genetic study demonstrated a homocygotic dinucleotide GT (GT) deletion at the beginning of exon 2 of the p47-phox gene. The same mutation has been found in European, American and Japanese patients. CONCLUSIONS: The molecular characterization of this pathology done for the first time in Chile is important for diagnostic classification, patient prognosis, and adequate genetic advice and a possible future therapy. PMID- 11008353 TI - [Treatment of hyperthyroidism with radioiodine: effects of administered dose on complications and thyroid function]. AB - BACKGROUND: Radio-iodine is a definite therapy for Graves disease hyperthyroidism. However, the optimal dosage is still debatable. AIM: To assess the effects of different radioiodine doses on thyroid function and complications in patients with hyperthyroidism. MATERIAL AND METHODS: A retrospective analysis of 139 patients with hyperthyroidism, treated with ratio-iodine between 1988 and 1998. Radio iodine dose used was classified as low (< 10 mCi), intermediate (10 14.9 mCi) or high (> or = 15 mCi). RESULTS: Thirty-five patients were treated with low doses, 33 with intermediate doses and 71 with high doses. There were no differences between these patients in age, disease severity, frequency of post treatment euthyroidism or complications. Patients treated with low doses had a higher frequency of persistent hyperthyroidism than patients treated with high doses (25.7 and 4.2% respectively, p < 0.001). Likewise, the frequency of subsequent hypothyroidism was 60% in patients treated with low doses and 84.5% of those with high doses, in whom it also appeared earlier. Associated complications were clinically irrelevant. In seven patients, Graves ophthalmopathy progressed after treatment, but this progression was not associated with the dose used. CONCLUSIONS: Radio iodine in high doses is useful, safe and effective for the treatment of Graves hyperthyroidism. PMID- 11008354 TI - [Triple therapy of short-term with azithromycin, amoxycillin and omeprazole for the eradication of Helicobacter pylori]. AB - BACKGROUND: The high cost and complexity of therapeutic schemes for the eradication of Helicobacter pylori has stimulated the search of simpler and cheaper treatment options. AIM: To evaluate the efficacy of 3 days of azithromycin 500 mg od, 7 days of amoxycillin 750 mg tid and omeprazole, 20 (Group A) or 40 mg (Group B) on randomization, as a treatment for Helicobacter pylori infection in patients with endoscopically diagnosed peptic ulcer. METHODS: H. pylori status of peptic ulcer patients was pathologically confirmed by the examination of five gastric biopsies using the Giemsa stain and by rapid urease testing in two gastric biopsies. H. pylori status was reassessed not less than 28 days after completing treatment. Adverse events and compliance were evaluated. RESULTS: Fifty four patients (29 men, 25 women, mean age 48 years) were enrolled, 28 in Group A and 27 in Group B. Per protocol the infection was cured in 58.8% of patients (30/51; 95% CI: 45-73%). On an intention to treat basis, H pylori infection was cured in 55%. Minor side effects including diarrhea and nausea were reported by 32% of patients. Ninety-five per cent of patients consumed more than 95% of prescribed medications. H. pylori was successfully eradicated in 61% of group A and 57% of group B patients (p = NS). CONCLUSION: Short term therapy with azithromycin was poorly effective in curing H. pylori infection. The compliance was excellent. Increasing Omeprazole from 20 to 40 mg/day did not improve treatment effectiveness. PMID- 11008355 TI - [BICAP tumor probe in the palliative treatment of malignant esophageal stenosis]. AB - BACKGROUND: BICAP tumor probe is a device that consists in an energy source and olives that deliver bipolar electricity. It can be used for the fulguration of esophageal tumors after endoscopic dilatation. AIM: To report the experience in the treatment of malignant esophageal stenoses using the BICAP tumor probe. PATIENTS AND METHODS: Patients with advanced esophageal tumors in aphagia, that were not candidates for palliative surgery were included in this study. After endoscopic dilatation, the tumor was fulgurated with the BICAP tumor probe. RESULTS: Twenty one patients (nine male, aged 43 to 91 years old) were treated with the device. A mean of 1.3 sessions with BICAP were necessary to obtain tumor permeabilization, which was obtained in all patients. One patient died of pneumonia 15 days after the procedure. All other patients were ingesting liquid or semisolid diets after two months of follow up. Mean survival after the procedure was 3.8 months. CONCLUSIONS: Electrical fulguration of esophageal tumors is a valid therapeutic alternative in aphagic patients. PMID- 11008356 TI - [Anorectal congenital malformations and their preferential associations. Experience of the Clinical Hospital of the University of Chile. Period 1979 1999]. AB - BACKGROUND: Anorectal atresia is a relatively frequent malformation in the newborn. According to the Latin American Collaborative Study for Congenital Malformations (ECLAMC), its frequency is 4.1 per 10,000 born alive. AIM: To determine the frequency of anorectal malformations at birth, and compare the figures with those of other maternity hospitals in Chile participating in ECLAMC, with the figures of the whole ECLAMC and with figures from other worldwide monitoring systems. PATIENTS AND METHODS: All births occurred in the University of Chile Clinical Hospital between January 1979 and August 1999, were reviewed. RESULTS: During the study period, 70,242 children were born, 4,486 had a malformation and 54 had an anorectal malformation (7.7 per 10,000 born alive). Fifty nine percent had other associated malformations (of the urinary tract in 42.5%, skeletal in 26% and cardiovascular in 18.5%). Five stillborn babies had other severe malformations. Twenty one children had a fistula. Forty three % were male, 39% female and 18% had ambiguous sex. When compared with normal controls, malformed newborns had a lower birth weight, lower gestational age and a higher mean maternal age, a higher frequency of metrorrhagia during the first trimester of pregnancy, a higher number of siblings with malformations and a higher degree of consanguinity among parents. CONCLUSIONS: The participation of recessive genes in the etiology of anorectal malformations is suggested. PMID- 11008357 TI - [Nocardia asteroides infection in a patient with systemic lupus erythematosus]. AB - Nocardia asteroides infection are unusually observed in systemic Lupus erithematous (SLE) patients. They are generally associated to steroidal and immunosuppressive therapy. We report a 24 years old female with SLE diagnosed in 1994 who developed a severe preeclampsia in her first pregnancy requiring emergency caesarean section. Post partum acute renal failure and type IV lupus nephropathy were treated with hemodialysis, methylprednisolone, cyclophosphamide and prednisone. Three months later, while she was receiving the fourth cyclophosphamide dose, she presented with a pleuro pneumonia and occipital abscess, both caused by Nocardia asteroides. She was treated with cotrimoxazole + cefixime and pleural decortication was required. Five months later, she developed Meningitis caused by Nocardia asteroides and hydrocephalus. She was treated with ceftriaxone, vancomycin, cotrimoxazole and ventricular shunting procedure. Two months later, a retroperitoneal abscess was diagnosed and surgically drained but the patient died, due to a methicillin-resistant Staphylococcus aureus septicemia. PMID- 11008358 TI - [Traumatic rupture of thoracic aorta and non operative treatment of concomitant splenic lesion. Case report]. AB - We report a 26 years old male that suffered a motorcycle accident resulting in a traumatic aortic rupture and splenic laceration. He was subjected to a surgical repair of the aortic lesion under complete heparinization. The splenic rupture was non operatively managed successfully. PMID- 11008359 TI - [Reflections on the current state of anatomic pathology in Chile]. AB - The present article reviews the most relevant aspects of the practice of Anatomic Pathology in Chile. The importance of biopsy and autopsy are analysed, in particular the consequences of the fall in the number of autopsies performed in our hospitals, in the recent decades. The modern pathologist is confronted with two situations: the renovation of the value of the autopsy and the acquisition of new diagnostic technologies, both not usually considered in the classical morphologic approach of Pathology. The ways to solve these problems in a discipline that is still a fundamental specialty in modern medicine are analyzed. PMID- 11008360 TI - [Tumorogenesis and mdm2 protein]. AB - Tumorogenesis is associated with several events by which a normal cell transforms itself into a tumour cell with an increased proliferation rate. One of the most important research initiatives in this area is the characterization of the molecular mechanisms involved in tumorogenesis and cancer. Oncogenes and tumour suppressor genes are directly involved in the cell cycle, differentiation, and apoptosis. The cellular oncogene MDM2 seems to be abnormally elevated in several human tumours, specially in sarcomas. The MDM2 gene product, mdm2 protein, pS3 and retinoblastoma (Rb) proteins, play crucial roles in the control of the cell cycle. The molecular interactions between mdm2, pS3 and Rb in cancer, are associated with a loss of control in the G1 phase of the cell cycle leading to uncontrolled cell proliferation. Studies by gene amplification appear to show an incomplete picture of mdm2 protein levels in tumour cells. The simultaneous determination of mdm2 protein and mRNA levels seems to give a more accurate interpretation of the abnormal function of the mdm2 protein. Thus, in addition to gene amplification, different mechanisms by which mdm2 is overexpressed in cancer cells also play an important role in tumorogenesis. PMID- 11008361 TI - [The terminally ill patient. Grupo de Estudios de Etica Cliinica de la Sociedad Medica de Santiago]. AB - The classification of a patient as terminally ill is based on an expert diagnosis of a severe and irreversible disease and the absence of an effective available treatment, according to present medical knowledge. Terminal diseases must not be confused with severe ones, since the latter may be reversible with an adequate and timely treatment. The physician assumes a great responsibility at the moment of diagnosing a patient as terminally ill. The professional must assume his care until the moment of death. This care must be oriented to the alleviation of symptoms and to provide the best possible quality of life. Also, help must be provided to deal with personal, legal and religious issues that may concern the patient. PMID- 11008362 TI - [Evidence based medicine]. PMID- 11008363 TI - The testicular descent in human. Origin, development and fate of the gubernaculum Hunteri, processus vaginalis peritonei, and gonadal ligaments. AB - Testicular descent has to be divided into the turn-out of the testis and epididymis from the abdomen proper and an inner abdominal descent of genital organs. Both events are closely related to and depend on the development and reorganisation of ligaments, mainly the gubernaculum Hunteri. These seemingly unambiguous events are controversially described since the first description of the gubernaculum, and results and specifics of other species were intermingled with data from humans, thus giving more confusion than lucidity in this important step of gonadal development. Here, we concentrate on human embryos, chronologically investigated by serial sections, scanning electron microscopy, three-dimensional reconstructions, microdissection and immunohistochemistry. The first question to be answered was whether a real inner descent of gonads occurs. We demonstrated this inner descent by showing the relations of the gonads, mesonephros, cranial mesonephric ligament and the anlage of the diaphragm with the vertebral segments. No explosion-like increase in certain vertebral segments was observed which might simulate a gonadal descent. The inner descent is coupled with the growth of the gonad (ovary or testis), the involution of mesonephros, the descending septum transversum or the anlage of the diaphragm, and the intercalated cranial mesonephric ligament. This ligament always inserts medially at the border between the gonad and mesonephros in close relationship to the abdominal ostium of the mullerian duct, a region where hydatides often occur. In contrast to the testes, the ovaries arrive very early--20-25 mm CRL--at their definitive position of S2/3 (level of linea terminalis pelvis), yet, are transversally oriented. The cranial gonadal ligament does not exhibit notable increase in length during inner descent. It does not contain blood vessels. While regressing in both sexes, it will be replaced by the plica formed by the ovarian vessels, that is the suspensorium ligament of the ovary as known in adults. The second point to be investigated was the origin, development, structure and fate of the gubernaculum Hunteri as well as the processus vaginalis peritonei. Their arrangement and composition is crucial for testicular descent. We discriminated five phases of their development and differentiation. Phase I characterises the early development of the gubernaculum of stage 14 CC (5-7 mm CRL) embryos. It arises as conus inguinalis and connects the abdominal wall lateral to the umbilical artery with the caudal part of the mesonephric fold. It is in this early stage that the localisation of the inner inguinal ring is defined. In phase II, stage 20-23 CC (21-30 mm CRL), three parts of the gubernaculum--abdominal, interstitial and subcutaneous--can be distinguished. The processus vaginalis peritonei appears with its dorsal layer firmly adhering to the ventral side of the gubernaculum. The gubernaculum inserts cranially into the mesenchyme of the genital ducts at their crossing-over. Opposite to it, but at some distance, a ligament connects the caudal pole of the testis with the dorsal mesenchyme of the genital ducts. In female embryos, the analge of the ovarian ligament appears as a U-shaped, double peritoneal fold. Phase II is subdivided in phase IIa (32-55 mm CRL), characterised by an enormous increase in length and volume of the gubernaculum and also an enlargement of the processus vaginalis peritonei. In phase III, sex-specific differences in gonadal position and gubernacular structure can be observed for the first time. Testes increase in volume and come close to the mesenchyme of the genital ducts. The caudal pole of the testis overlaps both ducts. We also subdivide this phase into phase IIIa (about 100 mm CRL) where two very important events occur in male foetuses: 1, the swelling of the gubernaculum, and 2, the gliding of the testis across the genital ducts. This gliding is permitted by both, the regression of the mullerian duct and t PMID- 11008364 TI - [The claw of the domestic cat (Felis catus)--analysis of its shape]. AB - The feline claws in the forepaw and hind paw are very different in shape. An objective method of comparison will be described. The differences in function and the factors with regard to the shape are discussed. PMID- 11008365 TI - Characterization of the swimming muscle of the anchovy Engraulis anchoita (Hubbs and Martini 1935). AB - The lateral muscle characteristics of individual female anchovies (Engraulis anchoita) were studied by histochemical and ultrastructural techniques. Red, pink and white fibres were discerned on the basis of the activity of the metabolic enzymes succinic dehydrogenase, phosphorylase and myofibrilar ATPase, and the usage of Sudan and periodic-acid Schiff staining techniques. Red fibres were located at the surface, white fibres deep inside and pink fibres in between. The red fibres appeared flattened or ribbon-shaped in transverse section and they were located in rows separated by myosepta of connective tissue. The red and pink fibres were multi-innervated whereas the white fibres were terminally innervated. The relative proportion of the red muscle increased toward the caudal region, the white muscle diminished toward that region, whereas the proportion of pink muscle did not vary according to region, being scarce throughout. The distribution of capillaries was also investigated, and as a result, the vascular supply proved to be scarce in both white and pink muscle. PMID- 11008366 TI - [Representation of the nerves of the wing of psittacines (Amazon, gray parrot} using a digital photographic technique]. AB - In four blue-fronted Amazon and five African grey parrots, the nerves innervating the muscles of the wings were carefully dissected and described. The preparation steps were recorded with a digital camera. This technique has now advanced so far that the image quality is almost the same as that of a normal camera. The pictures are stored in a computer. They can be enhanced and published via the Internet. PMID- 11008367 TI - Effects of retinoic acid exposure in utero on mouse vibrissal follicle development. AB - It is known that topical all-trans-retinoic acid (RA) modulates growth and differentiation of skin and its cutaneous appendages. To examine whether a pre natal exposure to a potentially non-teratogenic dosage of all-trans-RA had any effect on vibrissal follicle development, the histologic and immunohistochemical responses to RA during its morphogenesis in NMRI mouse were investigated. After a single oral dose of 30 mg/kg body weight of all-trans-RA on day 11.5 of gestation, no fetal malformations were detected and the histological features and the distribution of keratin (K) proteins in comparable stages of vibrissal development were similar for the untreated, vehicle-treated and RA-treated mice. The absence of teratogenic response and of adverse effects on the vibrissae under the experimental conditions indicates that this protocol may be useful for investigation of the effects of pre-natal exposure to RA on the post-natal development of experimental tumours in the mouse skin. PMID- 11008368 TI - Histochemical and morphometric aspects of the lateral musculature of different species of teleost marine fish of the Percomorphi order. AB - Cross-sections of the lateral musculature of 19 species of teleost fish of the Percomorphi order were studied histochemically and morphometrically. The red, pink and white muscles were observed in all the species except in Seriola dumerilii and Scomber japonicus, in which no pink muscle was found. The red fibres were the smallest due to their minimal diameters. Histochemically and according to their location at least five types were found in the same species: typical red, in rosette, of the border, infiltrated pink and in mosaic. The pink fibres were larger than the red but smaller than the white. The form and disposition of the pink muscle varied according to the species. This muscle is made up by fibres of medium size, moderate m-ATPase and intermediate oxidative activities. Their histochemical properties change near red and white muscles, where they acquire the histochemical characteristics of the typical red fibres or the low m-ATPase activity of white fibres, respectively. The white muscle is the widest muscular layer. All of the species, except Thalassoma pavo are made up by three fibre types (small with high m-ATPase activity, medium with moderate m ATPase activity, and large with low m-ATPase activity). They are mixed and show a histochemical and morphometric mosaic form. The extent of this mosaic depends on the species. In most of the analysed species it was observed in areas close to the pink muscle, but in three of them (Liza aurata, Seriola dumerilii and Coris julis) it occupied all of the myotome. PMID- 11008369 TI - Megakaryocytes and platelets in the spleen of the dromedary camel (Camelus dromedarius). AB - This study was carried out on spleens of clinically healthy camels (n = 35) of both sexes (0.5-15 years) by routine histology, electron microscopy and immunohistochemistry using 12 anti-bovine platelet antibodies. Megakaryocytes were observed in the red pulp. Their number decreased with age: they were found in the spleens of all camels under 8 years old but only in 57% of camels over 8 years old. Only two antibodies (IVA37 and IVA38) cross-reacted with camel platelets. A large number of platelets were found in the splenic cords and the marginal zone. Ultrastructurally, the platelets were oval in shape surrounded by a plasma membrane, and their cytoplasm was rich in glycogen and contained less dense granules. Microtubules and microfilaments were found at their periphery. Several platelets were observed in the red pulp. There are similarities in some surface antigens of bovine and camel platelets. The presence of megakaryocytes in the camel spleen indicates a thrombopoietic function of the spleen until adulthood but that this decreases with age thereafter. PMID- 11008370 TI - Growth-promoting effects of different fractions of extra-embryonic coelomic fluid on embryonic development. AB - In the early stages of embryonic development, many growth-promoting molecules must be provided by the maternal system. These factors may be supplied locally to the embryo, by the decidua, the placenta, or the yolk sac. In this study the growth-promoting potential of extra-embryonic coelomic fluid (EECF) and its fractions was investigated. The embryonic requirement of growth-promoting molecules may be studied by reducing the growth-supporting capacity of serum. Thus, ultrafiltration of rat serum was carried out for 8 h using Millipore filters with a molecular weight exclusion of 30 kDa. Rat embryos at 9.5 days of age were cultured for 8 days for anembryonic yolk sacs, and then EECF was collected and divided into three different molecular weight fractions by ultrafiltration. Rat embryos were cultured for 48 h in whole rat serum and the serum retenate (which has low growth-supporting capacity) in the presence and absence of EECF, its fractions, or in EECF only. Embryos grown in retenate showed severe growth retardation, and the addition of EECF significantly improved embryonic growth. The fraction which contained the molecules with molecular weight between 10 and 30 kDa had significantly more effect on embryonic development than the other fractions. This fraction of EECF was analysed by gel electrophoresis. Three of the four protein bands observed in this fraction were identified by amino-terminal sequencing as alpha-fetoprotein precursor (22 kDa), apolipoprotein A1 precursor (24 kDa) and fetal haemoglobin Y2 chain (14 kDa), none of which are likely to be responsible for the growth-promoting activity. To further investigate growth-promoting proteins, EECF was Western-blotted to nitrocellulose membranes and probed with antisera against rat prolactin, epidermal growth factor, insulin-like growth factors I and II and human placental lactogen. No immunoreactive bands were detected in the EECF, suggesting that either these proteins are not present or are present at levels too low to be detected. Although the growth-promoting effect of the EECF was demonstrated in this study, the molecules responsible remain uncharacterized. PMID- 11008371 TI - Temperature effect on muscle growth of the axial musculature of the sea bass (Dicentrarchus labrax L.). AB - In order to determine the temperature effect on the axial muscle growth of sea bass, a stock of larvae was subjected to the following incubation and cultivation temperatures, respectively: 15 degrees C/ambient, 15/17 degrees C, 17 degrees C/ambient and 17/17 degrees C. In all groups the cross-sectional area of white and red muscles and the number and average area of the white and red muscle fibres were quantified. Results showed that the embryonic period, pre-larval phase and the end of metamorphosis were accelerated at higher temperatures. During the endogenous feeding period, muscle growth took place by fibrillar hypertrophy, and was not influenced by the temperature. Thereafter (external feeding) muscular hyperplasia began, and growth of all the muscular parameters was favoured by the effect of high incubation and cultivation temperatures, with the latter having higher influence. High incubation temperature had an slight effect on muscle growth and body length, which was only observed from 15 days. Metamorphosis finished at 3 +/- 0.4 cm in all the larvae, but this length was earlier reached at higher temperatures. At 120 days, the largest growth was obtained in the larvae maintained at a higher temperature. PMID- 11008372 TI - On the morphology of the liver in the two-humped camel (Camelus bactrianus). AB - The liver of a two-humped camel (Camelus bactrianus) was examined by means of gross anatomy and histology. The liver appeared characteristically as an enlarged triangle in visceral aspect, and showed the following lobation: the left lateral and medial lobes, the right lobe, the caudate lobe and the quadrate lobe. These findings were similar to those for the one-humped camel. Histologically, a distinct lobulation, a typical arrangement of the interlobular connective tissue, and the trias was confirmed. PMID- 11008373 TI - Has active immunization against gonadotrophin-releasing hormone any effect on testis innervation in the pig? An immunohistochemical study. AB - The innervation of porcine testes was studied in intact animals and in boars undergoing active immunization against gonadotrophin-releasing hormone (GnRH) by means of immunohistochemistry using antibodies to tyrosine hydroxylase (TH), dopamine beta-hydroxylase (D beta H), vasoactive intestinal polypolypeptide (VIP), neuropeptide Y (NPY), synaptosome-associated protein of 25 kDa (SNAP-25) and protein gene product 9.5 (PGP 9.5). Moreover, the distribution of luteinizing hormone (LH) receptors in clusters of Leydig cells was also investigated. To identify these cells easily, either the NADPH-diaphorase histochemical technique or the Mayer counter-staining procedure was applied. Differences in the distribution pattern and relative density of particular subsets of intratesticular nerve fibres were observed in immunized boars as compared to those found in the intact animals. In the testes of non-treated animals, only single TH-immunoreactive (TH-IR) nerve fibres were observed. However, many D beta H-IR nerve terminals surrounded blood vessels in the tunica albuginea and parenchyma. Very scarce VIP-IR nerves occurred only in the tunica albuginea, mainly in close vicinity to blood vessels. Immunoreactivity to NPY occurred in single nerve fibres. Immunoreactivity to SNAP-25 and PGP 9.5 was found in single nerve fibres distributed mainly in the tunica albuginea. The interstitial cells were heavily stained for LH-receptors and NADPH-diaphorase. In the testes of immunized animals, only single TH-IR nerve fibres, scattered mainly in the tunica albuginea, were observed. Some TH-IR nerve terminals were also encountered in the parenchyma of the organ, where they were always associated with blood vessels. D beta H-IR nerve fibres formed a dense network distributed throughout the testis in association with the capsule, vasculature and interstitium. Some fibres were observed to run between seminiferous tubules. VIP-IR nerve fibres were located in the neighbourhood of blood vessels in the tunica albuginea and parenchyma. Only single VIP-IR nerves were found between seminiferous tubules. Numerous NPY-IR nerve fibres occurred in the tunica albuginea and parenchyma of the organ. SNAP 25-IR and PGP 9.5-IR nerve terminals formed a dense network distributed throughout the testis and many fibres were observed between seminiferous tubules. Interstitial cells were very weakly stained for LH receptors or NADPH-diaphorase. PMID- 11008374 TI - beta-Lactam derivatives as enzyme inhibitors: 1-peptidyl derivatives of 4 phenylazetidin-2-one as inhibitors of elastase and papain. AB - N-Peptidyl substituted azetidin-2-ones were synthesized and evaluated as inhibitors of the serine protease elastase, and the cysteine protease papain. All compounds were synthesized from 4-phenylazetidin-2-one, either from the racemate or from the pure enantiomers. The (S)-enantiomer was prepared by enantioselective synthesis from (S)-beta-phenyl-beta-alanine, while the (R)-enantiomer was obtained by enzymatic resolution with alpha-chymotrypsin. N-Alkylation with bromoacetates introduced a spacer group which, after hydrolysis to the free acid, was acylated with amino acid esters or di- or tripeptide esters. The enzymatic assays proved some derivatives to be effective inhibitors of PPE and/or papain. N BOC protected amino acid derivatives without a spacer group inhibited PPE reversibly, while derivatives with spacer group showed either weak or no inhibitory properties. On the other hand, papain was inactivated irreversibly by ethyl (RS)-2-oxo-4-phenylazetidin-1-acetate. The highest inhibitory activity against papain was found for the diastereomers of N-(2-oxo-4-phenylazetidin-1 acetyl)-L-alanyl-L-valine benzyl ester, a compound with a spacer group. PMID- 11008375 TI - A qualitative model for the histamine H3 receptor explaining agonistic and antagonistic activity simultaneously. AB - A pharmacophore model for histamine H3 ligands is derived that reveals the putative interaction of both H3 agonists and antagonists with an aspartate residue of the receptor. This interaction is determined by applying the density functional theory implemented in a program package adapted for parallel computers. The model reveals a molecular determinant explaining efficacy as the conformation of the aspartic acid residue differs according to whether it is binding to agonists or antagonists. The differences in structure-activity relationships (SAR) observed for the lipophilic tails of different classes of H3 antagonists are now explained, since the model reveals two distinct lipophilic pockets available for antagonist binding. PMID- 11008376 TI - Synthesis and anticonvulsant activity of acetylenic quinazolinone derivatives. AB - Acetylenic derivatives of quinazolinones and quinazolinediones were synthesized and evaluated for their anticonvulsant activity. Most compounds displayed seizure antagonizing activity in the maximal electroschock test (MES test) in most cases associated with little or no acute neurotoxicity determined in the rotorod test. Only three compounds exhibited significant activity in the seizure threshold test with subcutaneous pentylenetetrazole (scMet test). Based on the ED50 in the MES test, 1,3-bis-(prop-2-ynyl)-quinazoline-2,4-(1H,3H)-dione(9a) was about ten-fold less active than phenytoin or carbamazepine but about as active as mesuximide. PMID- 11008377 TI - A novel series of 2-carboxytetrahydroquinolines provides new insights into the eastern region of glycine site NMDA antagonists. AB - A series of potent 4-substituted tetrahydroquinolines has been synthesized and biologically tested in order to refine the eastern region of the pharmacophore model for glycine site NMDA antagonists concerning the assessment of lipophilicity, flexibility, and hydrogen bonding. Displacement studies on rat cortical membranes using [3H]-5,7-dichlorokynurenic acid as a radioligand indicated that binding affinities are markedly enhanced when additional hydrogen accepting groups are introduced into the eastern region of the 2 carboxytetrahydroquinolines. Among the most potent ligands were some urea, sulfonylurea, and crown ether compounds as interesting leads for new diagnostics, especially for the evaluation of PET tracers, which allow biodistribution studies and NMDA receptor studies in the living organism. PMID- 11008379 TI - Training. A key issue for the future of general practice. PMID- 11008378 TI - A new flavonoid from Limonium axillare. AB - A new flavonol glycoside identified as myricetin 3-O-beta-D-sorboside (1) has been isolated from the leaves of L. axillare. The new compound showed a moderate inhibition of Ehrlich ascites carcinoma cells. PMID- 11008380 TI - Aboriginal health. Can leadership and innovation make a difference? Part 2. PMID- 11008381 TI - Medial forefoot pain. PMID- 11008382 TI - Not every child who coughs has asthma. PMID- 11008383 TI - Irritable bowel syndrome. Practical management. AB - BACKGROUND: Irritable bowel syndrome (IBS) is a common problem affecting one in six Australians. In any patient complaining of chronic or recurrent abdominal pain, chronic diarrhoea or constipation or bloating, IBS should top the list of possibilities. OBJECTIVE: To outline an approach to the practical management of patients with IBS. DISCUSSION: A positive diagnosis can be made if a few simple questions are asked, and the diagnosis can be confirmed by a small number of tests to exclude structural disease. Improved understanding of the differences in gut function in patients with IBS is likely to result in the development of some exciting new therapies. PMID- 11008384 TI - Rectal bleeding. AB - BACKGROUND: Rectal bleeding, although common, is frequently alarming to patients because of its association with bowel cancer. An understanding of the causes of rectal bleeding and a systematic approach to diagnosis provides the basis for preventive measures and appropriate treatment. OBJECTIVE: This article discusses the causes, history, examination and investigation of rectal bleeding according to the age of the patient. Treatment of the various conditions is outlined. DISCUSSION: Although haemorrhoids are the commonest cause of rectal bleeding, most patients over 40 years old presenting with this symptom require colonoscopy in order to screen for and treat premalignant polyps and colorectal cancer. PMID- 11008385 TI - Coeliac disease. Update 2000. AB - BACKGROUND: Coeliac disease is increasingly recognised as a cause of nutritional disturbance in the Australian community. Diagnosis may be delayed because the presentation is highly variable. OBJECTIVE: To outline an approach to diagnosis and management of coeliac disease. DISCUSSION: Newer, more specific serological testing is helpful in screening asymptomatic individuals but endoscopic small intestinal biopsy is required for diagnostic certainty. A gluten free diet induces remission but not cure. Lifelong adherence reduces the risk of complications, including malignancy. All patients diagnosed with coeliac disease require expert dietetic assessment and counselling and all should be encouraged to join their state Coeliac Society. Excellent internet based sites now offer appropriate information for patients and their families. PMID- 11008386 TI - Anal fissures. AB - BACKGROUND: Anal fissures are common conditions, presenting with bleeding, itching, and pain of varying severity. Pain and bleeding is frequently attributed to haemorrhoids, which may delay commencement of appropriate therapy. Other causes for bleeding must also be excluded, with investigations taking into account the clinical findings and the age of the patient. OBJECTIVE: To discuss the pathogenesis and management of anal fissures. DISCUSSION: Recent studies have changed our understanding of the pathophysiology of anal fissures. It is now known that the majority of fissures are caused by internal sphincter spasm and resulting in ischaemia of the anal mucosa. Pharmacological agents that relax the sphincter have provided a novel approach to treatment, allowing surgery to be avoided in some patients. PMID- 11008388 TI - Casting acute fractures. Part 1--Commonly asked questions. AB - BACKGROUND: Correct splinting of acute fractures offers comfort and is often the definitive treatment for many simple injuries. The use of a cast is a common method of splinting. The best methods for application of casts and their rationale are poorly understood by many doctors and not detailed well in standard texts. OBJECTIVE: This series will attempt to address some common initial casting techniques suitable for general practice. Specific time and cost saving manoeuvres will be given. Common pitfalls will be identified. DISCUSSION: The primary care physician traditionally applies the first cast. This coincides with the time of greatest risk as fractures are potentially unstable, and swelling is often anticipated rather than actually present. The swelling and associated morbidity from a cast can be considerable. Optimising comfort, function and safety in the initial cast is multifactorial. Attention to these factors gives a far better short and long term outcome for the patient. This also reduces medicolegal risk and the cost of unplanned return visits due to unnecessary morbidity. PMID- 11008387 TI - COX-2 inhibitors. What is their place? AB - BACKGROUND: Nonsteroid anti-inflammatory drugs (NSAIDs) are widely used in Australia for the treatment of arthritis and the relief of pain. However, side effects, particularly those affecting the gastrointestinal tract, are of considerable cost to the individual and the community. OBJECTIVE: To examine the role of selective cyclo-oxygenase-2 (COX-2) inhibitors in the anti-inflammatory armamentarium. DISCUSSION: COX-2 inhibitors are of similar clinical efficacy to nonselective NSAIDs. They cause significantly fewer endoscopic ulcers, but these lesions are usually clinically asymptomatic. Dyspepsia rates are 1-2% lower. The frequency of serious gastrointestinal complications (symptomatic ulceration, bleeding, perforation) appears to be reduced by 0.5-1.0%. As these problems occur infrequently, routine use of COX-2 inhibitors would not be cost effective. Although no data are yet available, patients at high risk of NSAID induced gastrointestinal complications may be the group for whom COX-2 inhibitors will provide most clinical benefit. PMID- 11008389 TI - Patient education. Caring for your plaster. PMID- 11008390 TI - Acute sore throat and bronchitis. How effective are antibiotics? PMID- 11008391 TI - Sudden loss of vision in an elderly woman. PMID- 11008392 TI - Sudden onset of breathlessness. What are the possibilities in the pregnant patient? PMID- 11008393 TI - Midfoot pain. PMID- 11008395 TI - Screening for bilharzia. PMID- 11008394 TI - Pulmonary embolism. PMID- 11008396 TI - To be or not to be. Evidence in support of contemplative practices--Part 1. PMID- 11008397 TI - Is it all black magic? PMID- 11008398 TI - Methodological/ethical issues and general practice research. PMID- 11008399 TI - The end of full time general practice. PMID- 11008400 TI - Chinese attitudes to institutional care of their aged. A study of members from the Chung Wah Association, Western Australia. AB - OBJECTIVE: To examine Chinese attitudes to institutional care of their aged and to identify the factors that influence these attitudes. METHOD: A cross sectional survey of 815 randomly selected members of the Chung Wah Association, Western Australia was undertaken in 1997 using a mailed self administered questionnaire. The survey instrument was developed through a literature review, interviews and pilot testings and consisted of categorical and noncategorical items. The data were analysed using the SAS (V 6.12) software package. Factor analysis, logistic regression and Chi-square analysis were used on the data. The statistical significance level was set at p < or = 0.05. RESULTS: A total of 431 (53%) completed responses were received. The study showed that 86% of the respondents agreed that the disabled elderly should be institutionalised. Age contributed mainly to the respondents' attitudes to institutionalising the disabled elderly, while the respondents' sex, education, occupation, country of origin and length of residence in Australia did not. Social responsibilities together with the health factors of the elderly had important effects on these attitudes. About 55% of respondents agreed that their Chinese culture influenced their attitude. Only 21% of the respondents agreed that the nondisabled elderly should be institutionalised, 54% disagreed, while 24% remained neutral. Respondents who were likely to institutionalise their disabled elderly were also unlikely to live with their children when they became disabled. A majority of respondents were in favour of government funding of institutions and care givers of the elderly. CONCLUSION: The study provided several important observations and information for health planners and general practitioners to identify the aged at risk of institutionalisation and make arrangements for community support for effected families so that the aged can be encouraged to remain with their families. PMID- 11008401 TI - Are part timers real GPs? Attitudes of general practitioners toward those who work part time. AB - BACKGROUND: Current attitudes within the medical profession suggest that one cannot be a 'real' doctor unless one works full time. The aim of this study was to determine the views and attitudes of a sample of GPs toward part time practice. METHOD: As part of a larger study, GPs in two divisions in metropolitan Melbourne were sent a questionnaire based survey. GPs were asked to rate statements about part time practice using a Likert scale. RESULTS: The response rate was 55% (n = 246). The majority of GPs disagreed with the statement 'part time GPs are not real GPs' but feelings were mixed about whether part timers covered the same range of clinical content as full timers. One fifth of respondents questioned the competence of part time GPs. Male GPs, those over 55 years and full timers were more likely to hold negative views toward part time general practice. DISCUSSION: Our results show a 'generational' pattern toward part time practice. With the increasing numbers of women entering general practice, these attitudes will need to change to accommodate the fact that more GPs will be choosing to work part time in the future. PMID- 11008402 TI - Publicly-funded influenza and pneumococcal immunization programs in Canada: a progress report. PMID- 11008403 TI - [Bases and future perspectives of oncologic surgery in the integral treatment of advanced neoplasms]. PMID- 11008404 TI - [Neoadjuvant chemo-radiotherapy in the treatment of rectal cancer: preliminary results]. AB - Many studies in the literature have confirmed the role of combined therapy in the treatment of rectal neoplasms. Aim of the Authors' study was to evaluate the efficacy of preoperative neoadjuvant radiochemotherapy in these patients. The study group consisted of 15 patients, affected by rectal adenocarcinoma T < 4 M0 with age < 75 years, observed from January to August 1998, who underwent to RT of pelvis with three fields--total dose 32 Gy- in ten days associated with 5 FU(500 mg/m2) during the first five. After restaging the patients were operated on. Anterior resections (AR) were followed by mechanical colorectal anastomosis in 10 patients, coloendoanal anastomosis in 4, while in 1 case an abdominoperineal resection (APR) was performed. The protocol was well tolerated in every patient and perioperative complications were similar to those in the control group. A considerable reduction of the mass was obtained in a great percentage of patients studied, confirmed by pathologic examination like "down staging" of cancer. In no case a complete response to the therapy was observed (pT0N0). Survival in the patients operated on for rectal adenocarcinoma in about 50% and local relapse is the most important site of recurrent disease. The aim of neoadjuvant protocol is the control of lymphatic spread and reduction rectal neoplasm allowing coloanal anastomosis in the treatment of the 1/3 inferior rectal cancer. Literature data confirm a significant decrease of local relapse, probably with a better survival. The Authors study confirms that combined preoperative therapy is well tolerated in most of the cases and it represents the protocol of choice in the patient affected by rectal cancer. PMID- 11008405 TI - [Guidelines for routine assessment of cardiologic surgical risk: electrocardiogram]. AB - Cardiac complications are the main cause of perioperative mortality. A preoperative electrocardiogram and cardiologist's consultation are usually performed to get information about cardiac state of patients undergoing surgery and to prevent complications. In their study the Authors applied to 1715 patients undergoing surgery an evaluation schedule for the cardiac surgical risk, including an ECG as first-level test, performed systematically on the whole sample. The aim was to assess predictive value of this preoperative examination to verify its efficacy as routine test. Basing on results achieved, electrocardiogram is not routinely indicated before noncardiac elective surgery, but it should be requested for the patients having high risk of cardiac complications at an accurate clinical-anamnestic examination. Therefore, clinical judgement, that is the first level of any preoperative evaluation, should be the basis for ordering an ECG to be considered a second-level test. PMID- 11008406 TI - [Isolated cystic cervical lymphatic metastasis from occult papillary carcinoma of the thyroid: unusual and rather difficult diagnosis]. AB - Thyroid papillary carcinoma, exceptionally, can appear with cervico-lateral metastases in absence of thyroid evident disease. In that case we can have an infected branchial cyst, therefore is very important a correct preoperative diagnosis with FNAB. In literature were described 30 cases of cervico-lateral cystic masses that the histology revealed as occult papillary carcinoma metastases. Our experience is about 80 cases of papillary carcinoma, in only 2 cases the cancer was evident as a tight-elastic tumour in cervico-lateral region, hypoechogenic to the US scan with irregular and thick wall: one of the patient had multinodular goiter; the other one didn't have clinical of US scan character of thyroid disease. FNAB on lymph nodes shows doubtful cells evidence. Both patient had a total thyroidectomy with functional cervico-lateral and MAS lymphadenectomy. In one patient histology showed a papillary carcinoma of 4 mm in the right lobe, in the other patient the cancer was unknown. FNAB, in all doubtful cases, is the most important diagnosis test. PMID- 11008407 TI - [Surgical treatment of arterial hypertension: experience and results]. AB - Hypertension is a wide spread disease. In a small percentage of cases (nephrosclerosis, renovascular hypertension, endocrine hypertension) surgical treatment could be resolutive. The Authors describe their experience of the last five years about the subject, emphasize the technical detail taken and the results achieved; they outline the necessity to give the indications for surgery in wise manner considering clinical, laboratory and instrumental findings. PMID- 11008408 TI - [Total versus subtotal thyroidectomy for multiple node goiter: experience with 350 surgically treated cases]. AB - The surgical treatment of benign thyroid pathology and in particular the multinodular form is controversial. This controversy exists and one must choose either perform conservative surgical therapy such as lobectomy or sub-total thyroidectomy or total thyroidectomy. From January 1985 to July 1999, the Authors observed 534 cases of benign thyroid pathology. Of these 534, 330 were diagnosed with primitive goiter, and 20 with recurrent goiter; 13 cases of the primitive goiter type were found to have microfoci of carcinoma inside the goiter on the definitive histologic exam. There were 275 patients who underwent total thyroidectomy and the remaining 75 cases with ultrasound proven multinodular goiter were treated with lobectomy or sub-total thyroidectomy, depending on the appearance at the time of the operation as well as the macroscopic and microscopic exams of the integrity of the glandular parenchyma performed while in the operating room. The Authors demonstrated that in multinodular goiter there is a high percentage of cases with disease involving the entire gland. Many areas of apparently healthy tissue are found to be microscopically involved with a variable grade of chronic lymphocytic thyroiditis or follicular hyperplasia or even lobular dysplasia with the tendency to form nodules. These signs of disease of this organ tend to recur if not treated. PMID- 11008409 TI - Neck hibernoma: case report and literature review. AB - The case history of a 23 year old man with a large hibernoma of the cervical region is reported. Th tumour showed all the clinical and pathologic characteristic of these rare benign neoplasms. A review of the most recent literature supports the Authors' conclusion that local excision is curative. PMID- 11008410 TI - [Diaphragm rupture caused by closed trauma. A more and more frequent condition]. AB - The Authors report a case of rupture of the diaphragm in blunt trauma of the abdomen, to examine the more significant features of this condition whose incidence proves to be increasing, after a review of the literature data, parallel to the increase of accidents on the road and at work. They examine the most important pathogenic problems and these related to the mechanism leading to diaphragmatic lesion and they highlight the deeper pathophysiological changes suffered from the patient with such a condition. They draw attention to what they consider to be the most important aspect, and therefore the diagnosis. In fact, in accordance with the various series of cases reported in the literature, it is crucial in the diagnostic problem is when the possible cause of the rupture is suspected and necessary diagnostic cascade (Rx, echotomography, TC) is activated so to highlight the lesion. Therefore a quick diagnostic fitting is necessary for the survival of the acute patient with hernia of the thorax cavity, as well as quick surgical correction of the lesion that, as in the case described, serves to achieve the best possible anatomic-functional results. PMID- 11008411 TI - [Gastric epithelial dysplasia]. AB - Gastric epithelial dysplasia represents the only true histological marker of gastric cancer. In this bringing up to date, such subject is reproposed in consideration of taking into account the most recent acquisitions, subdividing gastric dysplasia into two degrees only: moderate and severe. For the first time an immunophenotypic study is made by means of the evaluation of gastric-entero pancreatic antigens, which better identify the evolutive potential of the two degrees of gastric dysplasia and, furthermore, the clinical development is evaluated, thus showing the necessity of a strict endoscopic surveillance of such lesion. PMID- 11008412 TI - Correlation between cytochrome P-450 system and liver function tests during experimental liver transplantation. AB - The aims of this investigation are: 1) to assess the function of the hepatocyte in transplanted porcine liver, immediately after reperfusion, by monitoring both LFTs and the MEGX levels; 2) to search for correlation between MEGX and LFTS, in an effort to evaluate the metabolic mechanisms occurring in the early liver transplantation revascularization phase. The MEGX test was found to be less than 50 micrograms/ml in all the recipients and all the LFTS tested have been reported to be out the normal range. Furthermore our data has shown a statistically significant correlationship between the MEGX values and those of alkaline phosphatase and prothrombin and a highly significant correlationship with cholinesterase. PMID- 11008413 TI - [Surgical indications in pulmonary tuberculosis]. AB - Since the begin of the century, the surgery for treatment of pulmonary tuberculosis showed an important evolution. The procedure has proved to be useful in the 20% of patients. It appears that the percentage is drug-resistant or complicated. The authors describe their experience in the years from 1992 to 1997 and, in accordance with the literature in this field, outline the good prognosis to 5 years, with a percentage of 90-96% of non-infected patients. Very important is the time of surgical intervention, the compliance of the medical treatment in the previous 6 months, excepting in the emergency, and the perfect obliteration of pleural cavity to avoid next reinfection and the development of broncopleural fistulas. PMID- 11008414 TI - Nonlinear Markov process amplitude EEG model for nonlinear coupling interaction of spontaneous EEG. AB - To develop an appropriate model for representing spontaneous electroencephalography (EEG) is an important and necessary work in the field of neuroscience. The Markov process amplitude (MPA) EEG model has been proposed in our previous work for representing the features of the EEG in terms of a few parameters. However, being a linear model, the linear MPA EEG model cannot perfectly describe the spontaneous EEG that displays nonlinear phenomena. In this paper, the nonlinear Markov process amplitude (nonlinear MPA) EEG model that includes nonlinear components is introduced. The consistent consideration of the nonlinear features of the EEG investigated by N. Wiener and P. L. Nunez can be seen from the nonlinear MPA EEG model. The similarity in the time domain and the goodness of fitting in the frequency domain with respect to the ongoing EEG are shown. As a result, the EEG power spectrum can be decomposed into the spontaneous components and the nonlinearly coupled components by use of the nonlinear MPA EEG model, which is useful for a better understanding the mechanism of the EEG generation. PMID- 11008415 TI - Threshold modeling of autonomic control of heart rate variability. AB - Even in the absence of external perturbation to the human cardiovascular system, measures of cardiac function, such as heart rate, vary with time in normal physiology. The primary source of the variation is constant regulation by a complex control system which modulates cardiac function through the autonomic nervous system. Here, we present methods of characterizing the statistical properties of the underlying processes that result in variations in ECG R-wave event times within the framework of an integrate-and-fire model. We first present techniques for characterizing the noise processes that result in heart rate variability even in the absence of autonomic input. A relationship is derived that relates the spectrum of R-R intervals to the spectrum of the underlying noise process. We then develop a technique for the characterization of the dynamic nature of autonomically related variability resulting from exogenous inputs, such as respiratory-related modulation. A method is presented for the estimation of the transfer function that relates the respiratory-related input to the variations in R-wave event times. The result is a very direct analysis of autonomic control of heart rate variability through noninvasive measures, which provides a method for assessing autonomic function in normal and pathological states. PMID- 11008416 TI - Adaptive noise cancellation in a multimicrophone system for distortion product otoacoustic emission acquisition. AB - This study focuses on adaptive noise cancellation (ANC) techniques for the acquisition of distortion product otoacoustic emissions (DPOAEs). Otoacoustic emissions (OAEs) are very low level sounds produced by the outer hair cells of normal cochleas, spontaneously or in response to sound stimulation as a byproduct of a frequency and threshold sensitivity increasing mechanism. Current OAE recording systems rely on test probe noise attenuation and synchronous ensemble averaging for increasing signal-to-noise ratios (SNRs). The efficiency of an ANC algorithm for noise suppression was investigated using three microphones: one placed in the test ear, one in the nontest ear for internal noise reference; one near the subject's head for external noise reference. The system proposed was tested with simulations, off-line averaging and real-time implementation of the ANC algorithm. Simulation results showed that the technique had a potential noise reduction capability of 24 dB for complex multifrequency noise signals. Off-line results were positive, with a mean SNR improvement of 4.9 dB. Real-time results indicated that the use of an ANC algorithm in combination with standard averaging methods can reduce noise levels by as much as 10 dB beyond that obtained with standard noise reduction methods and probe attenuation alone. PMID- 11008417 TI - An orthogonal least squares-based fuzzy filter for real-time analysis of lung sounds. AB - Pathological discontinuous adventitious sounds (DAS) are strongly related with the pulmonary dysfunction. Its clinical use for the interpretation of respiratory malfunction depends on their efficient and objective separation from vesicular sounds (VS). In this paper, an automated approach to the isolation of DAS from VS, based on their nonstationarity, is presented. The proposed scheme uses two fuzzy inference systems (FISs), operating in parallel, to perform the task of adaptive separation, resulting in the orthogonal least squares-based fuzzy filter (OLS-FF). By applying the OLS-FF to fine/coarse crackles and squawks, selected from three lung sound databases, the coherent structure of DAS is revealed and they are efficiently separated from VS. The important time domain DAS features, related to diagnostic information, are preserved and their true location and structural morphology are automatically identified. When compared to previous works, the OLS-FF performs quite similarly, but with significantly lower computational load, resulting in a faster real-time clinical screening of DAS. PMID- 11008418 TI - New aspects to event-synchronous cancellation of ECG interference: an application of the method in diaphragmatic EMG signals. AB - An "event-synchronous interference canceller" (ESC) for cancellation of electrocardiographic (ECG) interference in diaphragmatic electromyographic (EMGdi) signals is addressed in this paper. ESC pursues the concept of the "event synchronous adaptive interference canceller" (ESAIC), which was proposed in [1] as a specific application of the well known "adaptive noise canceller" (ANC) paradigm, but ESC uses a simple adaptive gain control (AGC) instead of the complex adaptive filter of the ANC. The proposed ESC method is evaluated using both computer simulations and real EMGdi data, and its efficiency in interference cancellation is compared to that of ESAIC. Of particular interest is the result that the ESC can replace the ESAIC providing better performance as well as a considerable reduction of computational costs. PMID- 11008419 TI - Analysis of a sleep-dependent neuronal feedback loop: the slow-wave microcontinuity of the EEG. AB - Increasing depth of sleep corresponds to an increasing gain in the neuronal feedback loops that generate the low-frequency (slow-wave) electroencephalogram (EEG). We derived the maximum-likelihood estimator of the feedback gain and applied it to quantify sleep depth. The estimator computes the fraction (0%-100%) of the current slow wave which continues in the near-future (0.02 s later) EEG. Therefore, this percentage was dubbed slow-wave microcontinuity (SW%). It is not affected by anatomical parameters such as skull thickness, which can considerably bias the commonly used slow-wave power (SWP). In our study, both of the estimators SW% and SWP were monitored throughout two nights in 22 subjects. Each subject took temazepam (a benzodiazepine) on one of the two nights. Both estimators detected the effects of age, temazepam, and time of night on sleep. Females were found to have twice the SWP of males, but no gender effect on SW% was found. This confirms earlier reports that gender affects SWP but not sleep depth. Subjectively assessed differences in sleep quality between the nights were correlated to differences in SW%, not in SWP. These results demonstrate that slow wave microcontinuity, being based on a physiological model of sleep, reflects sleep depth more closely than SWP does. PMID- 11008420 TI - Automatic detection of ST-T complex changes on the ECG using filtered RMS difference series: application to ambulatory ischemia monitoring. AB - A new detector is presented which finds changes in the repolarization phase (ST-T complex) of the cardiac cycle. It operates by applying a detection algorithm to the filtered root mean square (rms) series of differences between the beat segment (ST segment or ST-T complex) and an average pattern segment. The detector has been validated using the European ST-T database, which contains ST-T complex episodes manually annotated by cardiologists, resulting in sensitivity/positive predictivity of 85/86%, and 85/76%, for ST segment deviations and ST-T complex changes, respectively. The proposed detector has a performance similar to those which have a more complicated structure. The detector has the advantage of finding both ST segment deviations and entire ST-T complex changes thereby providing a wider characterization of the potential ischemic events. A post processing stage, based on a cross-correlation analysis for the episodes in the rms series, is presented. With this stage subclinical events with repetitive pattern were found in around 20% of the recordings and improved the performance to 90/85%, and 89/76%, for ST segment and ST-T complex changes, respectively. PMID- 11008421 TI - The effect of anisotropy on the potential distribution in biological tissue and its impact on nerve excitation simulations. AB - We present a finite difference solution of the potential distribution associated with electrical current stimulation in an anisotropic in-homogeneous tissue environment and compare it to the isotropic case. The results demonstrate that there can be significant errors associated with the assumption of isotropic tissue properties in calculating the potential distribution along an axon in nerve excitation simulations. These errors can have a significant impact on predicted nerve fiber recruitment patterns when evaluating the efficacy of specific surface or intramuscular stimulus electrode configurations. The results of this study also suggest when a more comprehensive tissue model should be implemented in an electrode design study. Simulation results indicate that the isotropy assumption is worst under bipolar electrode stimulation as opposed to monopolar stimulation and that the bipolar error increases as the distance between electrodes decreases. In light of these results, it is concluded that in order to avoid large errors in the calculated potential distribution along an axon, the isotropy assumption should only be used when the transverse depth from the electrode to the nerve is relatively small. PMID- 11008422 TI - Pacemaker interference and low-frequency electric induction in humans by external fields and electrodes. AB - The possibility of interference by low-frequency external electric fields with cardiac pacemakers is a matter of practical concern. For pragmatic reasons, experimental investigations into such interference have used contact electrode current sources. However, the applicability to the external electric field problem remains unclear. The recent development of anatomically based electromagnetic models of the human body, together with progress in computational electromagnetics, enable the use of numerical modeling to quantify the relationship between external field and contact electrode excitation. This paper presents a comparison between the computed fields induced in a 3.6-mm-resolution conductivity model of the human body by an external electric field and by several electrode source configurations involving the feet and either the head or shoulders. The application to cardiac pacemaker interference is also indicated. PMID- 11008423 TI - A statistical mechanical analysis of postural sway using non-Gaussian FARIMA stochastic models. AB - In this paper, postural sway is modeled using a fractional autoregressive integrated moving average (FARIMA) family of models: the center-of-pressure (COP) motion is viewed in terms of a self-similar, anti-persistent random-walk process, obtained by fractionally summating non-Gaussian random variables, whose correlation structure for small time lags is shaped by a linear time-invariant low-pass filter. The model parameters are: the strength of the stochastic driving, e.g., the root mean square (rms) value of the time-difference COP motion; the DC gain, damping ratio and natural frequency of the filter; the Hurst exponent, which measures the random-walk antipersistence magnitude. In the proposed modeling procedure, a graphical estimator for determining the Hurst exponent is cascaded to a method for matching autoregressive (AR) models to fractionally difference COP motion via higher order cumulants. The effect of the presence or absence of vision on the model parameter values is discussed with regard to data from experiments on healthy young adults. PMID- 11008424 TI - Functional reentry's influence on intracellular calcium in the LRd membrane equations. AB - This paper examines relationships between transmembrane potential (Vm), [Ca2+]i dependent membrane ionic currents, and [Ca2+]i handling by the sarcoplasmic reticulum (SR) in a two-dimensional model of cardiac tissue. Luo-Rudy dynamic (LRd) membrane equations were used because they include detailed formulations for triggered SR Ca2+ release dependent on membrane Ca2+ influx (CICR) and for spontaneous SR Ca2+ release following calsequestrin buffer overload (SCR). Reentry's rapid rate (110-ms cycle length) elevated [Ca2+]i and limited CICR, which in turn promoted SCR that occurred at intervals of 320-350 ms, was preferential at sites located inside the functional center, and destabilized the reentrant activation sequence. Although adjustment of LRd parameters for SR Ca2+ modified SCR interval and peak [Ca2+]i in voltage clamp simulations with a command waveform representing Vm time course within the functional center, SCR persisted. Using the same command waveform, SCR also occurred with an alternate SR Ca2+ formulation that represented subcellular details underlying CICR. LRd parameter adjustments to promote CICR and limit SCR in subsequent reentry simulations failed to eliminate SCR completely, as they modulated SCR intervals in a manner consistent with the voltage clamp simulations. Taken together, our findings support a destabilizing influence of functional reentry on [Ca2+]i handling. However, [Ca2+]i instabilities did not always fractionate depolarization wavefronts during reentry. Fractionation depended, in part, upon CICR and SCR parameters in the LRd formulation for SR Ca2+ release. PMID- 11008425 TI - The effect of geometric and topologic differences in boundary element models on magnetocardiographic localization accuracy. AB - This study was performed to evaluate the changes in magnetocardiographic (MCG) source localization results when the geometry and the topology of the volume conductor model were altered. Boundary element volume conductor models of three patients were first constructed. These so-called reference torso models were then manipulated to mimic various sources of error in the measurement and analysis procedures. Next, equivalent current dipole localizations were calculated from simulated and measured multichannel MCG data. The localizations obtained with the reference models were regarded as the "gold standard." The effect of each modification was investigated by calculating three-dimensional distances from the gold standard localizations to the locations obtained with the modified model. The results show that the effect of the lungs and the intra-ventricular blood masses is significant for deep source locations and, therefore, the torso model should preferably contain internal inhomogeneities. However, superficial sources could be localized within a few millimeters even with nonindividual, so called standard torso models. In addition, the torso model should extend long enough in the pelvic region, and the positions of the lungs and the ventricles inside the model should be known in order to obtain accurate localizations. PMID- 11008426 TI - Paired MEG data set source localization using recursively applied and projected (RAP) MUSIC. AB - An important class of experiments in functional brain mapping involves collecting pairs of data corresponding to separate "Task" and "Control" conditions. The data are then analyzed to determine what activity occurs during the Task experiment but not in the Control. Here we describe a new method for processing paired magnetoencephalographic (MEG) data sets using our recursively applied and projected multiple signal classification (RAP-MUSIC) algorithm. In this method the signal subspace of the Task data is projected against the orthogonal complement of the Control data signal subspace to obtain a subspace which describes spatial activity unique to the Task. A RAP-MUSIC localization search is then performed on this projected data to localize the sources which are active in the Task but not in the Control data. In addition to dipolar sources, effective blocking of more complex sources, e.g., multiple synchronously activated dipoles or synchronously activated distributed source activity, is possible since these topographies are well-described by the Control data signal subspace. Unlike previously published methods, the proposed method is shown to be effective in situations where the time series associated with Control and Task activity possess significant cross correlation. The method also allows for straightforward determination of the estimated time series of the localized target sources. A multiepoch MEG simulation and a phantom experiment are presented to demonstrate the ability of this method to successfully identify sources and their time series in the Task data. PMID- 11008427 TI - Simultaneous optical and nuclear magnetic resonance spectroscopy for monitoring cardiac energetics in vivo. AB - There are a number of applications in which it is useful to simultaneously collect data from what are traditionally separate instrumentation modalities. In particular, in vivo physiological investigations in which data from parallel experiments must be correlated would benefit from simultaneous data collection through 1) elimination of subject variability, 2) elimination of treatment variability, and 3) a reduction in the number of animal preparations required. Here we describe the simultaneous collection of fluo-3 optical fluorescence and 31P nuclear magnetic resonance (NMR) spectra to measure intracellular calcium levels and high-energy phosphate metabolism, respectively, in vivo. This work is part of ongoing research into the profound anoxia tolerance exhibited by the hearts of certain turtle species. An NMR compatible optical fluorescence spectrometer was constructed and tested. In the 31-cm bore of a 2 T superconducting magnet, NMR and optical spectra were collected every 10-15 min from the in situ, in vivo hearts of anesthetized turtle subjects prior to and during one to three hours of anoxia. It was found that while PCr stores became significantly depleted during anoxia, beta-adenosine triphosphate (ATP) levels remained within 20% of control values, and intracellular diastolic calcium levels did not vary by more than 10%. The ability to make simultaneous phosphorus and calcium measurements on a single subject is important to understanding the exact relationship between phosphorus energy state and maintenance of calcium homeostasis. PMID- 11008428 TI - In vitro simultaneous measurement of refractive index and thickness of biological tissue by the low coherence interferometry. AB - We proposed and demonstrated in vitro simultaneous measurement of refractive index and thickness of biological tissue. The technique is based on the low coherence interferometry combined with precise translation stages. Refractive indices were determined with the accuracy of less than 1% for tissue samples of a few hundred micron thickness, including chicken tissue, human tooth and nail. Simultaneous measurement of refractive index and thickness of multilayer tissue are also demonstrated. PMID- 11008429 TI - A maximum-likelihood base caller for DNA sequencing. AB - The procedures used to sequence the human genome involve the electrophoretic separation of mixtures of dioxyribonucleic acid (DNA) fragments tagged with reporting groups, usually fluorescent dyes. Each fluorescent pulse which arrives from an optical detector corresponds to a nucleotide (base) in the DNA sequence, and the subsequent process of base detection is known as base calling. Generating longer and more accurate sequences in the base-calling process will reduce the high cost of DNA sequencing. This paper presents an automated base-calling algorithm, referred to as maximum-likelihood base caller (MLB), which is based on maximum likelihood equalization for digital communication channels. Based on 125 experimental datasets, MLB averaged up to 40% fewer errors than the widely used ABI base caller from the Applied Biosystems Division of PE Corporation. MLB's accuracy rivaled that of another well-known base caller, Phred, surpassing it on datasets with high background noise. PMID- 11008430 TI - An improved configuration for the reduction of EMG in electrode cuff recordings: a theoretical approach. AB - A theoretical investigation of different electroneurogram recording techniques using electrode cuffs is presented. A new screened tripole arrangement is proposed with a higher inherent signal to interference ratio than the true tripole, which also allows the nulling of the residual electromyogram signal. The reduction in interference is small because the electrode impedance is large compared to the source resistance. PMID- 11008431 TI - How electrode size affects the electric potential distribution in cardiac tissue. AB - We investigate the effect of electrode size on the transmembrane potential distribution in the heart during electrical stimulation. The bidomain model is used to calculate the transmembrane potential in a three-dimensional slab of cardiac tissue. Depolarization is strongest under the electrode edge. Regions of depolarization are adjacent to regions of hyperpolarization. The average ratio of peak depolarization to peak hyperpolarization is a function of electrode radius, but over a broad range is close to three. PMID- 11008432 TI - Sensitivity and versatility of an adaptive system for controlling cyclic movements using functional neuromuscular stimulation. AB - This study evaluated an adaptive control system (the PG/PS control system [2]) that had been designed for generating cyclic movements using functional neuromuscular stimulation (FNS). Extensive simulations using computer-based models indicated that a broad range of control system parameter values performed well across a diverse population of model systems. The fact that manual tuning is not required for each individual makes this control system particularly attractive for implementation in FNS systems outside of research laboratories. PMID- 11008433 TI - Selecting the corner in the L-curve approach to Tikhonov regularization. AB - The performance of two methods for selecting the corner in the L-curve approach to Tikhonov regularization is evaluated via computer simulation. These methods are selecting the corner as the point of maximum curvature in the L-curve, and selecting it as the point where the product of abcissa and ordinate is a minimum. It is shown that both these methods resulted in significantly better regularization parameters than that obtained with an often-used empirical Composite REsidual and Smoothing Operator approach, particularly in conditions where correlated geometry noise exceeds Gaussian measurement noise. It is also shown that the regularization parameter that results with the minimum-product method is identical to that selected with another empirical zero-crossing approach proposed earlier. PMID- 11008434 TI - Using evidence-based practice for managing clinical outcomes in advanced practice nursing. AB - Preparation of advanced practice nurses to assume leadership positions for clinical decision making requires that traditional ways of solving clinical problems be augmented with information from relevant, research-derived evidence. In this article, the authors describe how one graduate program prepares advanced practice nurses to use the best scientific evidence with clinical expertise to influence patient outcomes. The assignments that students complete in their program provide examples of evidence-based practice that apply quality improvement principles and science-based nursing interventions to create best practices. PMID- 11008435 TI - Satisfaction with managed care. AB - This article reports the findings of 1996, 1997, and 1998 patient satisfaction surveys administered to managed care enrollees in Utah. More than 14,000 managed care enrollees (both Medicaid and commercial) were selected randomly and contacted by telephone. The 38-question survey was based on Health Plan Employer Data and Information Set (HEDIS) and the National Committee for Quality Assurance (NCQA) measures. Demographic differences between the commercial and Medicaid population were identified. Medicaid enrollees were found to be higher users of health care services. Individuals reporting the greatest health plan satisfaction tended to be healthier. However, Medicaid enrollees reported greater overall health plan satisfaction than commercial enrollees. PMID- 11008436 TI - Linking nursing care interventions with client outcomes: a community-based application of an outcomes model. AB - This article describes a conceptual model that provides an organizing framework for assessing client outcomes in community-based settings. The "Outcomes Model for Community-Based Settings" (OMCBS) examines the relationships among structure, process, and outcomes juxtaposed to the dimensions of client, provider, and setting including sample measures for each variable. OMCBS incorporates the Omaha System including its comprehensive list of client health problems, nursing interventions, and an outcome rating scale assessing client knowledge, behavior, and health status to standardize nursing care and client outcomes. The OMCBS provides a strategy to evaluate the effectiveness of nursing interventions for clients in a variety of milieus. PMID- 11008437 TI - Critical care admissions criteria in community based hospitals: a pilot study with implications for quality management. AB - Critical care nurses are increasingly vulnerable to crises of decision-making. Lack of enforcement or vague admission guidelines in many critical care units- including physician coverage and time limits for newly admitted patients--might lead to quality of care concerns. Managers struggle with staffing shortages, decreasing resources to provide high-technology care, and in many instances the lack of administrative support to enforce criteria for utilization of critical care beds. This pilot study addresses quality of care concerns and other issues that are a great source of frustration for critical care nursing staff. PMID- 11008438 TI - Medication error reporting: a survey of nursing staff. AB - The objective of this article is to describe findings from a medication error (ME) survey, to estimate the extent of ME underreporting by comparison of survey results with written incident reports (IRs), and to determine factors associated with IR reporting of MEs. Participants were registered nurses from the 38-bed infant unit of a pediatric hospital. Most recent ME in each of four stages of the medication process was classified as to: timing, nature, whether the error was prevented from the patient, patient injury, and completed IR. Surveys were administered to nurses during mandatory skills session and were compared with IRs for MEs for the previous 6 months. The survey response rate was 93.5 percent; 72 nurses described 177 errors, 40.3 percent observed an ME in the previous week, 62.1 percent were prevented from reaching the patient and the likelihood of prevention was reduced in the later stages of the medication process. About 30 percent of MEs resulted in IRs. Administration errors were more likely to result in IRs compared with ordering errors, especially when the error was not prevented from the patient. There were 51 IRs for MEs. A multivariate logistic regression with completed IRs as the dependent variable showed a decreased likelihood of IRs for ordering than administration errors. IRs were more likely for wrong medication or dose errors and IRs were less likely for errors prevented from reaching the patient. The study found that by augmenting IR reporting of MEs and classifying errors by stage, anonymous ME surveys can be used for monitoring and guiding improvements to hospital medication systems. PMID- 11008439 TI - Performance improvement through clinical research utilization: the linkage model. AB - Performance improvement offers tremendous opportunities for research utilization in the clinical setting. The linkage model of research utilization is characterized by a user system that accesses a resource system, adapting research findings to clinical practice. A performance improvement consultant can be the link between these systems. Research application can be instrumental in improving care delivery as demonstrated in a project concentrating on the management of individuals experiencing a cerebrovascular accident. PMID- 11008440 TI - Nonlinear dynamics: chaos and catastrophe theory. AB - Nonlinear relationships are attracting much attention in nursing. Nonlinear dynamics are used abstractly as theory and also may be applied in a concrete sense to explain data. There is often confusion and misunderstanding about aspects of nonlinear dynamics. This article explores two of the more well known areas of nonlinear dynamics--chaos and catastrophe theory. A summary of each is given, areas of commonality and distinction are explored, and guidelines for application are identified. PMID- 11008441 TI - Tubular contents of equine dentin: a scanning electron microscopic study. AB - The dentinal tubules of 20 permanent equine incisors were investigated by scanning electron microscopy. Occlusal surfaces and longitudinal fracture planes of both etched and undecalcified teeth were examined. Three different types of structure were observed inside the dentinal tubular lumen. Odontoblastic processes could only be visualized in the circumpulpal parts of the tubules. The more peripheral parts were empty or housed cylindrical structures that probably correspond to the laminae limitantes. Collagen fibres were frequently observed in the tubular lumina and were most numerous in the circumpulpal parts of the tubules. PMID- 11008442 TI - Inheritance of disc calcification in the dachshund. AB - The occurrence of intervertebral disc calcification was investigated by conventional spinal radiography in eight families of wirehaired dachshunds, with each family comprising one sire, two dams and one litter from each dam. Each offspring was examined radiographically once at 24-35 months of age. The occurrence of disc calcification was rated according to four different scales. A strong correlation was found in the occurrence of disc calcification between offspring and mean parent (P < 0.001) and between offspring and dams (P < 0.005) on an either/or scale. Statistically significant estimates of heritability of 0.60 and 0.87 were found based on the offspring-sire relationship using the total score and three-class scale, respectively. Higher correlation estimates were found based on the dam-offspring relationship than based on the sire-offspring relationship, suggesting an effect of maternal environmental factors. PMID- 11008443 TI - Cardiopulmonary side-effects and pharmacokinetics of an emulsion of propofol (Disoprivan) in comparison to propofol solved in polysorbate 80 in goats. AB - The aim of this study was to determine whether any pharmacokinetic or pharmacodynamic differences exist in goats between propofol in its currently licensed form (Disoprivan) and a new 1% solution of propofol (NSP) containing polysorbate 80. Nine goats received, on two different occasions in a randomized double-blinded order, 4 mg/kg propofol intravenously (i.v.; Disoprivan or NSP). To detect differences in cardiopulmonary effects and pharmacokinetics, the Wilcoxon signed rank test for paired data was used. In the NSP group the duration of initial apnoea was significantly longer, and 6 and 12 min after drug application PaO2 levels were significantly lower than in the Disoprivan group. Mean cardiovascular parameters did not differ significantly between the groups but in the NSP group in six goats marked changes in blood pressure occurred: systolic arterial pressures fell to a minimum of 40-60 mmHg within the first 10 min. This was followed by a marked increase in blood pressure, with maxima exceeding 300 mmHg. In the NSP group the half-life of propofol was significantly longer, the clearance rate was smaller and the areas under the drug concentration time curves were larger than in the Disoprivan group. The cardiopulmonary side effects of NSP suggest that propofol dissolved in polysorbate 80 is not a suitable alternative to the current formulation of propofol. PMID- 11008444 TI - Clinical experiences of treating septic arthritis in the equine by repeated joint lavage: a series of 39 cases. AB - The condition of septic arthritis was treated in 12 foals with 21 affected joints (Group I) and in 27 adult horses. The adult horses were divided into three groups, based on aetiology of the condition: haematogenous (Group II, n = 6), iatrogenic (Group III, n = 6), and perforating trauma (Group IV, n = 15). The treatment consisted of an initial systemic antibiotic that anticipated the microbial agents that were considered most likely per group, repeated through-and through joint lavages every other day and non-steroidal anti-inflammatory drugs. The antibiotics were adjusted to the results of bacteriological culture and susceptibility tests. Joint lavages were continued until the white blood cell count dropped below 15 G/l and bacteriological culture was negative, after which a single dose of a short-acting corticosteroid was administered intra articularly. Joint recovery rate in group I was 71%. Patient recovery rate of the foals, however, was lower (42%). Three foals were killed for reasons other than arthritis; one foal because of an arthritis-related problem and three foals because of persistent arthritis. Overall joint recovery rate, equalling patient recovery rate, in the adult horses was 81%. The expected predominance of Streptococcus spp. in haematogenous arthritis in adult horses was not confirmed, indicating that in these cases also, an initial antibiotic treatment with a broad spectrum combination is preferable. It is concluded that with intensive treatment, the prognosis of septic arthritis in the adult horse can be classified as fair to even good. Results in the foals are not as good, but this seems to be more due to the specific problems surrounding the equine neonate than to unresponsiveness to the treatment. PMID- 11008445 TI - Effect of calcium supplements to a maize-soya diet on the bioavailability of minerals and trace elements and the accumulation of heavy metals in growing rats. AB - Thirty-five (five groups with seven animals each) male albino rats (initial average weight = 44 g) were fed phytate-rich diets (analysed phytic acid concentration = 6.9 g/kg) based on maize and soy bean meal (5 g Ca, 3 g P, 1.2 g Mg, 23 mg Zn, 10 mg Pb, 5 mg Cd/kg diet). Experimental diets were supplemented with 0, 2, 4, 6 and 8 g calcium from CaCO3 per kg. The supplementation of increasing amounts of calcium resulted in a dose-dependent decrease in the apparent absorption of phosphorus. Furthermore, apparent zinc absorption and femur zinc concentration were moderately decreased due to the calcium supplementation. Kidney Cd concentration was significantly lower in rats that were fed the high calcium diets in comparison with the control animals. Femur lead concentration and hepatic delta-aminolevulinic acid dehydratase, which are known to be sensitive parameters of lead accumulation remained unchanged by the different dietary treatments. Magnesium absorption as well as liver and plasma zinc concentration and activity of plasma alkaline phosphatase were also unaffected. Although calcium supplementation may lead to a decrease in the accumulation of certain heavy metals such as cadmium, the carry-over of lead was not affected under the given experimental conditions. Furthermore, calcium phytate-zinc interactions may adversely affect zinc bioavailability in growing rats. PMID- 11008446 TI - Stability of tetracycline in water and liquid manure. AB - The objective of this study was to establish a suitable model for the prediction of the environmental stability of antibiotics in water and liquid manure. The model consists of incubation systems for solutions with installations for stirring ventilation. It was used to examine the stability of tetracycline in ventilated and unventilated Ringer's solution and liquid pig manure for a period of 8 days. Tetracycline concentrations decreased significantly during the experiment. The fastest degradation was observed in ventilated liquid manure. The test might be used as a screening test for the determination of the degradation half-life of antibiotics in the first phase of an environmental safety study according to directive 92/18/EEC. PMID- 11008447 TI - [Lawyers and journalists]. PMID- 11008448 TI - [Ketolides]. PMID- 11008449 TI - [Arthrosis and arthritis]. PMID- 11008450 TI - [Common colds and their complications]. PMID- 11008451 TI - [Nose drops. Applications, indications, alternatives]. PMID- 11008452 TI - Pepper spray just won't go away. PMID- 11008453 TI - Medical malpractice: 'fixing' is not so simple. PMID- 11008454 TI - Public health, safety, and financial problems posed by domestic violence (DV) PMID- 11008455 TI - Physician assistants. A young profession celebrates the 35th anniversary of its birth in North Carolina. PMID- 11008456 TI - Brainwashing and battering fatigue. Psychological abuse in domestic violence. AB - Intimate partner violence occurs often in the United States; it involves an interrelated combination of physical, sexual, and psychological abuse, usually directed against women. The psychological aspect deserves special attention because victims who lose their independence, self-esteem, and dignity tend to remain in abusive situations. The abuse is perpetrated by a domestic partner to maintain power and control in the relationship. To assert control, the abuser uses "brainwashing tactics" similar to those used on prisoners of war, hostages, or members of a cult. Common features of brainwashing include isolation, humiliation, accusation, and unpredictable attacks. The abusive environment produces real and anticipated fear, which contributes to the battered woman's belief that her situation is hopeless and that she must depend on her abuser. She develops coping strategies to deal with her oppressive environment, but eventually exhibits symptoms of "battering fatigue," similar to the battle fatigue of soldiers in combat who, like battered women, live in fear of being killed or severely injured. Recognizing the state of mind of these women can help us understand why it is difficult for them to flee their traumatic environment and why they may resort to suicide or homicide. For healthcare providers to screen and treat their patients adequately, it is imperative that they appreciate the complex and devastating psychological aspects of domestic violence. PMID- 11008457 TI - The doctor as witness. How good medical records can help victims of domestic violence, minimize time in court, and maximize effective testimony. PMID- 11008458 TI - Domestic violence and children. A review. PMID- 11008459 TI - Emergency medical services personnel and domestic violence. PMID- 11008460 TI - Lethal domestic violence in eastern North Carolina. AB - Strategies for preventing domestic violence can be tailored to a particular geographic or socioeconomic area if the patterns of domestic violence in the area are known. National statistics, although widely available, may not be applicable to a specific region. We reviewed homicide deaths in Eastern North Carolina between 1978 and 1999 to identify patterns in this rural area. Approximately 20% of the homicide deaths in eastern North Carolina are caused by intimate partners. Women accounted for 53% of the victims in 1976, similar to national figures but not rising to 72% as seen nationally in 1998. Latinos are an increasing presence in the area, but had only one recorded episode of lethal violence against an intimate partner. Gunshots accounted for most of the deaths (59% in men, 72% in women). Knowledge of such patterns can assist in selecting prevention strategies for this particular area. Over the last 25 years increasing attention has been devoted to domestic violence (DV), initially defined as abuse committed against a spouse, former spouse, fiancee, boy- or girlfriend, or cohabitant. As time has passed, the definition has been broadened to include other family members- elders, children, and siblings. The Centers for Disease Control and Prevention (CDC) now uses the term "intimate partner violence" for intentional emotional or physical abuse inflicted by a spouse, ex-spouse, a present or former boy- or girlfriend, or date. For the purposes of this paper, we consider DV interchangeable with intimate partner violence. There has been a national concern that abusive events are under-reported. The National Crime Victimization Survey, an anonymous household survey, indicated nearly 1 million incidents of non-lethal intimate partner violence per year between 1992 and 1996. The number decreased from 1.1 million in 1993 to 840,000 in 1996. Attempts to validate such data for a given geographic area often require subjects to violate anonymity--this may account for lower reports of violence. A recent national report from the Justice Department found a decline in both lethal and non-lethal DV. The number of men murdered by wives or girlfriends "plunged 60% from 1976 through 1998". FBI data on homicides showed that "intimate partners committed fewer murders each year during 1996, 1997, and 1998 than in any other year since 1976". Nationally, intimate partners caused 3000 deaths in 1976, 1590 (53%) in women; in 1998, they caused 1830 deaths, 1320 (73%) in women. But fatal cases of DV are only the tip of the iceberg, and may pertain only to a particular geographic area. We undertook the present study to assess the prevalence of lethal domestic violence in the 29 counties of eastern North Carolina (ENC) that make up the catchment area for the University Health Systems of Eastern Carolina (UHSEC). UHSEC includes the Brody School of Medicine at East Carolina University (BSM-ECU; previously known as East Carolina University-SOM) and Pitt County Memorial Hospital. PMID- 11008461 TI - Elder mistreatment. A guide for medical professionals. PMID- 11008462 TI - [Photodynamic therapy in pneumology]. PMID- 11008463 TI - [Effect of ambroxol on surfactant secretion and synthesis in isolated type II alveolar cells]. AB - The effect of ambroxol is attributed in part to an effect on surfactant synthesis and secretion. Evidence supporting this hypothesis is largely indirect; a direct effect of ambroxol on surfactant synthesis and secretion remains to be demonstrated. In this study a direct effect of ambroxol was evaluated using isolated alveolar type II cells. Secretion of labelled phosphatidylcholine was measured following the addition of increasing concentrations (10(-8) M to 10(-4) M) of ambroxol to the culture medium for increasing time intervals. There was no significant increase in surfactant secretion with increased ambroxol concentration or prolonged exposure time. Uptake of 3H-choline and synthesis into 3H-phosphatidylcholine was analyzed as an indicator of surfactant synthesis. Again, increasing concentrations of ambroxol (10(-7) M to 10(-5) M) were followed for 1, 2, 4, 8 and 24 h. There was no significant effect on synthesis at any time point. Concentrations higher than those mentioned here resulted in LDH release from cultured cells. Ambroxol which also has anti-oxidative and anti-inflammatory effects does not exhibit direct stimulatory effects on surfactant synthesis and secretion in isolated (rat) alveolar type II cells as has been demonstrated e.g. for beta 2-adrenergic stimulation. A specific pharmacologic way to stimulate the surfactant system remains to be developed. PMID- 11008464 TI - [Indications for corticosteroid therapy and sarcoidosis and the assessment of the results]. PMID- 11008465 TI - [Protective vaccination against pneumococci]. PMID- 11008466 TI - [Methods for the costing process in the field of economic evaluation of a rehabilitation program for patients with chronic obstructive lung diseases]. AB - Studies in health economics especially economic evaluations of health care technologies and programmes are getting more and more important. However, in Germany there are no established, validated and commonly used instruments for the costing process. For the economic evaluation of a rehabilitation programme for patients with chronic lung diseases such as asthma and chronic bronchitis we developed methods for identification, measurement and validation of resource use during the inpatient rehabilitation programme and during the outpatient follow-up period. These methods are based on methodological considerations as well as on practical experience from conducting a pilot study. With regard to the inpatient setting all relevant diagnostic and therapeutic resource uses could be measured basing on routine clinical documentation and validated by using the cost accounting of the clinic. For measuring the use of resources during the follow-up period in an outpatient setting no reliable administrative data are accessible. Hence, we compared a standardised retrospective patient questionnaire used in a 20-minute interview (n = 50) and a cost diary for the continuing documentation by the patient over a period of 4 weeks (n = 50). Both tools were useful for measuring all relevant resource uses in sufficient detail, but because of higher participation rates and lower dropouts the structured interview appears to be more suitable. Average total costs per month were 1591 DM (interview), respectively 1867 DM (cost diary). Besides productivity loss, costs for medication and GP visits caused the relatively highest resource uses. Practicable instruments were developed for the costing process as part of an economic evaluation in a German rehabilitation setting for pulmonary diseases. After individual modification, these could also be used for different indications and in other institutional settings. PMID- 11008467 TI - [Individualized, modular structured patient behavioral training in obstructive airway diseases during inpatient rehabilitation]. PMID- 11008468 TI - [COPD--a diagnostic and therapeutic challenge. Congress report on the workshop of Boehringer Ingelheim Pharma KG on March 3 20000 in Hamburg in the framework of the 20th congress of the German Society of Pneumology]. PMID- 11008469 TI - [Long-term oxygen therapy--from whom and how? Results of expert discussion in the framework of Pneumology Workships "COPD--an inventory at the beginning of the 21st century by Boehringer Ingelheim Pharma KG, Ingelheim on January 21 2000 in Seefeld, Tyrol]. PMID- 11008470 TI - [An invasive bronchoscopic strategy in the treatment of patients with suspected ventilator-associated pneumonia is not a superior clinical strategy]. PMID- 11008471 TI - lGradation of sideropneumoconiosis. AB - Pathological-anatomical analyses of biopsy and autopsy samples of 43 men with anamnestically established occupational exposure to welding fumes showed in 38 specimens (88%) characteristic alterations of varying degrees of sidero pneumokonioses. Based on recurrent histological findings of increase and activation of iron storing macrophages (siderophages) and varying degrees of fibrosis, sidero pneumokonioses are classified into three grades. Intraindividually and also toporegionally different degrees may well be present. Based on morphological criteria of the three grades, seven samples were classified as grade I, 21 specimens as grade II, and 10 samples as grade III. The results show that exposure to welding fumes may well cause pulmonary alterations reaching an impairment level, depending on length, extent and special circumstances of the respective exposure. In cases of so-called welders lungs, an obliging statement in an expert's opinion on the degree of functional impairment, however, can only be given when histological findings, detailed information on the occupational history and clinical functional parameters are combined. PMID- 11008472 TI - [Role of surgical procedures in the diagnosis and therapy of tuberculosis]. AB - Over the last decades the annual number of surgical intervention for treatment of pulmonary tuberculosis has steadily declined. Despite effective antituberculotic medication, there are still indications for thoracic surgery. The aim of the present study was to give an account of the diagnostic and therapeutic role of surgery of pulmonary tuberculosis. Therefore we analysed all operated patients with pulmonary tuberculosis between 1988 and 1999. In 2% of all patients with pulmonary tuberculosis surgical intervention was performed (n = 35). Their main indication was resection for suspected carcinoma, multi-drug resistance or non compliance to the medical treatment, decortication because of cavern rupture or empyema, hemoptoe and destroyed lung. A additional indication for surgery was pulmonary aspergilloma caused by superinfection of postspecific cavities. Postoperative complications like bleeding (> 1000 ml) or broncho-pleural fistula occurred in 37%, none of the patients died. Surgery is still a valid option fur the treatment of pulmonary tuberculosis. Before surgical intervention, a long term antituberculotic chemotherapy is essential. Patients have to be selected individually because of an increased morbidity. In 10% of all operated patients (n = 1132) with preoperative unidentifiable pulmonary mass the infiltrate was specific aetiology. PMID- 11008473 TI - [The state of tuberculosis in Germany 1998]. PMID- 11008474 TI - [Aspects of tuberculosis]. PMID- 11008475 TI - [nCPAP therapy does not affect respiratory mechanics of patients with sleep related respiratory disturbances]. AB - We investigated in patients with sleep disordered breathing the influence of long term nCPAP-therapy on airway mechanics and capillary blood gases. 109 patients (19 women, 90 men, age [mean +/- standard deviation] 59 +/- 10 years, apnea hypopnea-index 34 +/- 24/h) were treated for 27 +/- 33 months, with a mean pressure of 7.7 +/- 2 mbar, mean usage 5.6 +/- 1.9 h/night. Vital capacity, FEV1, thoracic gas volume and capillary blood gases did not change during therapy, irrespective of the smoking history. However, in a subgroup of 6 patients who were hypoxemic before therapy, there was a significant increase of PaO2 from 63 +/- 4 to 70 +/- 6 mm Hg (p < 0.01). Neither the height of the pressure nor the duration of therapy did influence the course of lung function data. PMID- 11008476 TI - [The driving simulation test "carsim" for assessing vigilance. Effect of driving practice and other factors in health subjects and in patients with sleep apnea syndrome]. AB - Among other factors, obstructive sleep apnoea (OSAS) patients suffer from impaired continuous attention. For registration and objectification as well as observation of the course of therapy, driving simulation programmes are particularly suitable. "Carsim", a driving simulation newly developed by us, simulates a bendy road via a screen where a vehicle is supposed to be kept on the right lane by operating a steering wheel. Apart from examination of quality criteria (objectivity, reliability, validity) and establishment of standard values, the influence of significant variables (age, gender, school education, profession, duration of disorder, AHI, BMI, visual and motor functionability, driving license, driving experience, Epworth score, accident frequency) on patients and healthy controls requires to be evaluated. For this purpose, 100 healthy controls, 200 OSAS patients and 30 patients under nCPAP were analysed. In healthy persons and patients under nCPAP, driving experience had no impact on the driving simulation results. OSAS patients with no driving license and no driving experience achieved lower marks for tracking than patients with driving license or a lot of driving experience (no driving license: x = 8058 +/- 10,878 track deviations; with driving licence: x = 2111 +/- 6564 track deviations; p < .001). According to our findings, untreated OSAS severely interferes with patients' attention capacity as well as their coping strategies. PMID- 11008477 TI - [From physiopathology to new developments in pharmacotherapy of bronchial asthma- a look into the future]. PMID- 11008478 TI - [Isolated tumor cells in bone marrow predicts reduced survival in lymph node negative non-small-cell lung cancer]. AB - BACKGROUND: It became recently evident that isolated tumor cells undetectable by conventional tumor staging are frequently present in bone marrow of patients with apparently localized non-small cell lung cancer (NSCLC). The clinical relevance of this minimal hematogenous tumor cell dissemination is under vigorous debate. METHODS: For tumor cell detection in the bone marrow we used monoclonal antibody CK2 against the epithelial intermediate filament protein cytokeratin 18. The influence of a positive bone marrow finding on clinical outcome was studied in 139 patients with NSCLC postoperatively staged as pT1-4, pN0-2, M0, R0 after a median follow up of 66 months (48-74). FINDINGS: Cytokeratin-18-positive cells in bone marrow were demonstrated in 83 (59.7%) patients at the time of primary surgery and in 6 of 12 representative patients analyzed twice 3-18 months after surgery. In patients without histopathological lymph node metastases (pN0; n = 66) the occurrence of > or = 2 tumor cells in bone marrow at primary surgery was a strong and independent predictor for overall survival (p = 0.007) in univariate analysis. The multivariate analysis showed a 2.8 times increased risk for shorter survival in patients with disseminated tumor cells versus patients without such cells. Four of the six patients with a positive CK status after surgery developed a tumor recurrence 11-44 months after the operation, while in none of the patients with a negative bone marrow at all times intervals showed a tumor relapse. CONCLUSIONS: Minimal residual bone marrow involvement is an independent prognostic factor for overall survival in patients with node-negative NSCLC, which may help to identify patients in need of an adjuvant systemic therapy. The postoperative persistence or re-appearance of tumor cells in bone marrow indicates that these are not only shredded cells but rather represent true micrometastasis. PMID- 11008479 TI - [Recommendations on the diagnosis of bronchial carcinoma. German Pneumology Society]. PMID- 11008480 TI - [Effects of significantly increased perinatal risks on developmental status in adolescence]. AB - In a 13-years-catamnesis a sample of 50 children with severe perinatal risks was assessed with regard to their physical, cognitive and behavioral development; selective drop-outs were analysed. In accordance with the results of other prospective studies there are deficits in visual-motor functioning, higher rates of somatic complaints, social withdrawal and attention deficits; a large number of these children attend special schools. The study presents predicative relations between risk factors of birth-time and the developmental status at youth. In comparison with typical single risks (e.g. gestational age or respiratory distress syndrome) a complex index for the perinatal risk, the so called birth-optimality offers no substantial advantage. A significant influence of social factors is shown for school achievement. Children of fathers with higher educational level are attending schools of a higher grade. The combination of biological and social factors improves the reliability of developmental prognosis; the Mental Developmental Index of the Bayley Scales of Infant Development provides additional prognostic accuracy. For compensating developmental disadvantages especially parents with lower educational level should be informed about beneficial care. Even children of these families, who scored low in early cognitive assessments should receive early intervention or therapeutic support. PMID- 11008481 TI - [Differences in linguistic representation of 10- to 12-year-old boys of divorced and not divorced parents]. AB - Twenty-eight 10- to 14 years old boys from divorced families (separation of the parents was two years before the investigation) were compared with 26 10- to 14 years old boys from two-parent families on the basis of interviews about attachment related themes and daily situations, and by means of the California Child Q-Sort (CCQ). The interviews were analyzed with regard to coherence of language, emotional problems, reflectivity, supportive representation of their parents, verbal and nonverbal emotional openness and social network, as well as for personality variables. The boys of the divorced group did barely talk about relations, had problems in coping with emotional stress, and were less reflective about themselves and others. They felt insufficiently supported by their parents and did not show their emotions openly. They talked about themselves as dependent from their social environment and as less adaptive to new situations. These indications suggest an insecure attachment representation predominantly as a result of the parents' behavior leading to divorce (Table 1). The results show that separations and family disruptions, as a group effect, strongly affect pre adolescent boys' state of mind. A minority of boys from the parental divorce group, however, show secure mental representations, comparable to a majority in the control group. Tables 2-5 show, independent of parental divorce, the major differences between boys with secure and boys with insecure mental representations. PMID- 11008482 TI - [The General Depression Scale in diagnosis of adolescents]. AB - A German version of the Center for Epidemiological Studies Depression Scale (CES D) was used in the Zurich Epidemiological Study of Child and Adolescent Psychopathology (ZESCAP). Psychometric analyses revealed clear evidence of a central factor of depression as reflected by the CES-D. Due to good reliability, a total score of the CES-D should be used in clinical diagnostics. The first cross-sectional wave of assessment within ZESCAP found that gender, nationality (indigenous vs. migrants) and residence (urban vs. rural) are important determinants of the total score. The article provides norms for adolescents. The study of the validity of the CES-D warrants further research. PMID- 11008483 TI - [As index patient into therapeutic alternatives?--Children in systemic family therapy and counseling]. AB - Since systemic therapy and counseling approaches have become established in child guidance, and children and adolescent psychiatry the understanding for children in the therapeutic context has also changed. They are part of the family system who, with their symptoms, point to restricting reality constructions and interaction patterns which create suffering (index patients). They are often treated correspondingly in the family setting. On this background this article investigates the special situation of children in the practice of systemic family therapy. The few results from studies from practice research dealing with this subject are compared with system-theoretical basic assumptions. The results indicate that the children are only insufficiently integrated into the therapy sessions and are excluded partly. They often experience the family discussions as being adult-oriented and feel that their needs and views are considered only slightly. Possible causes for this may be expecting too much emotionally and/or cognitively especially from younger children with family-oriented methods (such as circular questioning) as well as deficits in the training of family therapists. The closing thoughts on the therapeutic implications from the presented results include suggestions on methodical approaches more suitable for children and a reflected therapeutic attitude (transparency, participation, respect) towards children. PMID- 11008484 TI - Cyclic nucleotide phosphodiesterases: relating structure and function. AB - Cyclic nucleotide phosphodiesterases (PDEs) comprise a superfamily of metallophosphohydrolases that specifically cleave the 3',5'-cyclic phosphate moiety of cAMP and/or cGMP to produce the corresponding 5'-nucleotide. PDEs are critical determinants for modulation of cellular levels of cAMP and/or cGMP by many stimuli. Eleven families of PDEs with varying selectivities for cAMP or cGMP have been identified in mammalian tissues. Within these families, multiple isoforms are expressed either as products of different genes or as products of the same gene through alternative splicing. Regulation of PDEs is important for controlling myriad physiological functions, including the visual response, smooth muscle relaxation, platelet aggregation, fluid homeostasis, immune responses, and cardiac contractility. PDEs are critically involved in feedback control of cellular cAMP and cGMP levels. Activities of the various PDEs are highly regulated by a panoply of processes, including phosphorylation events, interaction with small molecules such as cGMP or phosphatidic acid, subcellular localization, and association with specific protein partners. The PDE superfamily continues to be a major target for pharmacological intervention in a number of medically important maladies. PMID- 11008485 TI - Thyroid hormone regulation of apoptotic tissue remodeling: implications from molecular analysis of amphibian metamorphosis. AB - Organogenesis and tissue remodeling are critical processes during postembryonic animal development. Anuran metamorphosis has for nearly a century served as an excellent model to study these processes in vertebrates. Frogs not only have essentially the same organs with the same functions as higher vertebrates such as humans, but also employ similar organogenic processes involving highly conserved genes. Development of frog organs takes place during metamorphosis, which is free of any maternal influences but absolutely dependent on the presence of thyroid hormone. Furthermore, this process can be easily manipulated both in intact tadpoles and in organ cultures by controlling the availability of thyroid hormone. These interesting properties have led to extensive morphological, cellular, and biochemical studies on amphibian metamorphosis. More recently, the cloning of thyroid hormone receptors and the demonstration that they are transcription factors have encouraged enormous interest in the molecular pathways controlling tissue remodeling induced by thyroid hormone during metamorphosis. This article summarizes some of the recent studies on the mechanisms of gene regulation by thyroid hormone receptors and isolation and functional characterization of genes induced by thyroid hormone during Xenopus metamorphosis. Particular focus is placed on the remodeling of the animal intestine, which involves both apoptosis (programmed cell death) of larval cells and de novo development of adult tissues, and the roles of thyroid hormone induced genes that encode matrix metalloproteinases during this process. PMID- 11008486 TI - Role of S6 phosphorylation and S6 kinase in cell growth. AB - This article reviews our current knowledge of the role of ribosomal protein S6 phosphorylation and the S6 kinase (S6K) signaling pathway in the regulation of cell growth and proliferation. Although 40S ribosomal protein S6 phosphorylation was first described 25 years ago, it only recently has been implicated in the translational up-regulation of mRNAs coding for the components of protein synthetic apparatus. These mRNAs contain an oligopyrimidine tract at their 5' transcriptional start site, termed a 5'TOP, which has been shown to be essential for their regulation at the translational level. In parallel, a great deal of information has accumulated concerning the identification of the signaling pathway and the regulatory phosphorylation sites involved in controlling S6K activation. Despite this knowledge we are only beginning to identify the direct upstream elements involved in growth factor-induced kinase activation. Use of the immunosupressant rapamycin, a bacterial macrolide, in conjunction with dominant interfering and activated forms of S6K1 has helped to establish the role of this signaling cascade in the regulation of growth and proliferation. In addition, current studies employing the mouse as well as Drosophila melanogaster have provided new insights into physiological function of S6K in the animal. Deletion of the S6K1 gene in mouse cells led to an animal of reduced size and the identification of the S6K1 homolog, S6K2, whereas loss of dS6K function in Drosophila demonstrated its paramount importance in development and growth control. PMID- 11008487 TI - Molecular characterization of monoamine oxidases A and B. AB - Monoamine oxidase A and B (MAO A and B) are the major neurotransmitter-degrading enzymes in the central nervous system and in peripheral tissues. MAO A and B cDNAs from human, rat, and bovine species have been cloned and their deduced amino acid sequences compared. Comparison of A and B forms of the enzyme shows approximately 70% sequence identity, whereas comparison of the A or B forms across species reveals a higher sequence identity of 87%. Within these sequences, several functional regions have been identified that contain crucial amino acid residues participating in flavin adenine dinucleotide (FAD) or substrate binding. These include a dinucleotide-binding site, a second FAD-binding site, a fingerprint site, the FAD covalent-binding site, an active site, and the membrane anchoring site. The specific residues that play a role in FAD or substrate binding were identified by comparing sequences in wild-type and variants of MAO with those in soluble flavoproteins of known structures. The genes that encode MAO A and B are closely aligned on the X chromosome (Xp11.23), and have identical exon-intron organization. Immunocytochemical localization studies of MAO A and B in primate brain showed distribution in distinct neurons with diverse physiological functions. A defective MAO A gene has been reported to associate with abnormal aggressive behavior. A deleterious role played by MAO B is the activation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a proneurotoxin that can cause a parkinsonian syndrome in mammals. Deprenyl, an inhibitor of MAO B, has been used for the treatment of early-stage Parkinson's disease and provides protection of neurons from age-related decay. PMID- 11008488 TI - Squalene synthase: structure and regulation. AB - Squalene synthase (SQS) catalyzes the first reaction of the branch of the isoprenoid metabolic pathway committed specifically to sterol biosynthesis. Regulation of SQS is thought to direct proximal intermediates in the pathway into either sterol or nonsterol branches in response to changing cellular requirements. The importance of SQS in cholesterol metabolism has stimulated research on the mechanism, structure, and regulation of the enzyme. SQS produces squalene, a C30 isoprenoid, in a two-step reaction in which two molecules of farnesyl diphosphate are condensed head to head. Site-directed mutagenesis of rat SQS has identified conserved Tyr, Phe, and Asp residues that are essential for function. The aromatic rings of Tyr and Phe are postulated to stabilize carbocation intermediates of the first and second half-reactions, respectively; the acidic Asp residues may be required for substrate binding. SQS activity, protein level, and gene transcription are strictly and coordinately regulated by cholesterol status, decreasing with cholesterol surfeit and increasing with cholesterol deficit. The human SQS (hSQS) gene has an unusually complex promoter with multiple binding sites for the sterol regulatory element binding proteins SREBP-1a and SREBP-2, and for accessory transcription factors known to be involved in the control of other sterol-responsive genes. SREBP-1a and SREBP-2 require different subsets of hSQS regulatory DNA elements to achieve maximal promoter activation. Current research is directed at elucidating the precise contribution made by individual SREBPs and accessory transcription factors to hSQS transcriptional control. PMID- 11008489 TI - Yeast chromatin structure and regulation of GAL gene expression. AB - Yeast genomic DNA is covered by nucleosome cores spaced by short, discrete length linkers. The short linkers, reinforced by novel histone properties, create a number of unique and dynamic nucleosome structural features in vivo: permanent unpeeling of DNA from the ends of the core, an inability to bind even full 147 bp core DNA lengths, and facility to undergo a conformational transition that resembles the changes found in active chromatin. These features probably explain how yeast can maintain most of its genome in a transcribable state and avoid large-scale packaging away of inactive genes. The GAL genes provide a closely regulated system in which to study gene-specific chromatin structure. GAL structural genes are inactive without galactose but are highly transcribed in its presence; the expression patterns of the regulatory genes can account for many of the features of GAL structural gene control. In the inactive state, GAL genes demonstrate a characteristic promoter chromosomal organization; the major upstream activation sequence (UASG) elements lie in open, hypersensitive regions, whereas the TATA and transcription start sites are in nucleosomes. This organization helps implement gene regulation in this state and may benefit the organism. Induction of GAL expression triggers Gal4p-dependent upstream nucleosome disruption. Disruption is transient and can readily be reversed by a Gal80p-dependent nucleosome deposition process. Both are sensitive to the metabolic state of the cell. Induction triggers different kinds of nucleosome changes on the coding sequences, perhaps reflecting the differing roles of nucleosomes on coding versus promoter regions. GAL gene activation is a complex process involving multiple Gal4p activities, numerous positive and negative cofactors, and the histone tails. DNA bending and chromosomal architecture of the promoter regions may also play a role in GAL regulation. Regulator-mediated competition between nucleosomes and the TATA binding protein complex for the TATA region is probably a central aspect of GAL regulation and a focal point for the numerous factors and processes that contribute to it. PMID- 11008490 TI - A coordinated interplay: proteins with multiple functions in DNA replication, DNA repair, cell cycle/checkpoint control, and transcription. AB - In eukaryotic cells, DNA transactions such as replication, repair, and transcription require a large set of proteins. In all of these events, complexes of more than 30 polypetides appear to function in highly organized and structurally well-defined machines. We have learned in the past few years that the three essential macromolecular events, replication, repair, and transcription, have common functional entities and are coordinated by complex regulatory mechanisms. This can be documented for replication and repair, for replication and checkpoint control, and for replication and cell cycle control, as well as for replication and transcription. In this review we cover the three different protein classes: DNA polymerases, DNA polymerase accessory proteins, and selected transcription factors. The "common enzyme-different pathway strategy" is fascinating from several points of view: first, it might guarantee that these events are coordinated; second, it can be viewed from an evolutionary angle; and third, this strategy might provide cells with backup mechanisms for essential physiological tasks. PMID- 11008491 TI - Signal transduction pathways and the modification of chromatin structure. AB - Mechanical and chemical signaling pathways are involved in transmitting information from the exterior of a cell to its chromatin. The mechanical signaling pathway consists of a tissue matrix system that links together the three-dimensional skeletal networks, the extracellular matrix, cytoskeleton, and karyoskeleton. The tissue matrix system governs cell and nuclear shape and forms a structural and functional connection between the cell periphery and chromatin. Further, this mechanical signaling pathway has a role in controlling cell cycle progression and gene expression. Chemical signaling pathways such as the Ras/mitogen-activated protein kinase (MAPK) pathway can stimulate the activity of kinases that modify transcription factors, nonhistone chromosomal proteins, and histones. Activation of the Ras/MAPK pathway results in the alteration of chromatin structure and gene expression. The tissue matrix and chemical signaling pathways are not independent and one signaling pathway can affect the other. In this chapter, we will review chromatin organization, histone variants and modifications, and the impact that signaling pathways have on chromatin structure and function. PMID- 11008492 TI - RGS proteins: lessons from the RGS9 subfamily. AB - RGS proteins enhance the time resolution of G protein signaling cascades by accelerating GTP hydrolysis of G alpha subunits of heterotrimeric G proteins. RGS9-1, a photoreceptor-specific RGS protein, is the first vertebrate member of this sizeable family whose physiological function in a well-defined G protein pathway has been identified. It is essential for normal subsecond recovery kinetics of the light responses in retinal photoreceptors. Understanding this role allows RGS9-1 to serve as a useful model for understanding how specificity and regulation of RGS function are achieved. In addition to the catalytic RGS domain, shared among all members of this family, RGS9-1 contains several other domains, which are also found in a closely related subset of RGS proteins, the RGS9 subfamily. One of these domains, the G gamma-like (GGL) domain, has been identified as the attachment site for G beta 5 proteins, which act as obligate subunits for this subfamily. Results from RGS9-1 and other subfamily members suggest that specificity is achieved by cell type-specific transcription, RNA processing, and G beta 5-dependent protein stabilization. In addition, membrane localization via specific targeting domains likely plays an important role. PMID- 11008494 TI - [Will refractive surgery replace glasses?]. PMID- 11008493 TI - Regulation of mammalian cell membrane biosynthesis. AB - This review explores current information on the interrelationship between phospholipid biochemistry and cell biology. Phosphatidylcholine is the most abundant phospholipid and it biosynthesis has been studied extensively. The choline cytidylyltransferase regulates phosphatidylcholine production, and recent advances in our understanding of the mechanisms that govern cytidylyltransferase include the discovery of multiple isoforms and a more complete understanding of the lipid regulation of enzyme activity. Similarities between phosphatidylcholine formation and the phosphatidylethanolamine and phosphatidylinositol biosynthetic pathways are discussed, together with current insight into control mechanisms. Membrane phospholipid doubling during cell cycle progression is a function of periodic biosynthesis and degradation. Membrane homeostasis is maintained by a phospholipase A-mediated degradation of excess phospholipid, whereas insufficient phosphatidylcholine triggers apoptosis in cells. PMID- 11008495 TI - [Tuberculosis eradication: realist perspective or utopia?]. PMID- 11008496 TI - [Epidemiology and physiopathology of chronic venous leg diseases]. AB - Chronic venous disorders of the lower limbs include a wide range of clinical manifestations involving more than one half of the population in the industrial countries. Varicose veins have a multifactorial origin, linked to ageing, environment (life habits, pregnancies...) and heredity. Their development is related to an increased distensibility of the venous wall and to haemodynamic dysfunction secondary to valvular incompetence. Cutaneous trophic changes and leg ulcers are the final result of severe primary varicose disease or post-thrombotic disease. Elder people are mostly concerned. Skin disease results from ambulatory venous hyperpressure due to several types of reflux involving superficial, deep and perforator veins. Its development is linked to a cutaneous microangiopathy, the central element of which is a decreased capillary density. Venous symptoms are quite widespread and women are more concerned than men; they are also of a multifactorial origin, varicose veins being only one of the causative factors. Their pathogenesis remains unclear, involving venous stasis and probably a functional insufficiency of the lymphatic system. PMID- 11008498 TI - [Primary varices of the legs]. AB - Varicose veins are a very common reason for medical referral. Diagnosis is made on clinical examination. The disease is generally benign, but can be complicated by cutaneous ulcers. Duplex ultrasound give informations on the superficial, deep and perforating venous systems. Treatment is not straightforward. Recurrences are common after surgery and sclerotherapy. Medical treatment consists in compression hosiery and symptomatic drugs. PMID- 11008497 TI - [Heavy and swollen legs]. AB - Painful sensation of heavy or swollen legs are non-specific symptoms frequently associated with chronic venous insufficiency. Clinical evaluation is the first step in defining the cause of the complaint and offering adequate treatment. When a heavy or swollen leg is associated with oedema, venous insufficiency, lymphatic or systemic disease must be considered. If symptoms occur during walking a vascular or nervous disease must be suspected. Associated erythema suggests infection (erysipelas). If clinical data are the cornerstone of diagnosis, difficulty may arise from the high frequency of superficial venous insufficiency and the readiness of linking too quickly any non specific complaint to this particular venous disease. PMID- 11008499 TI - [Drugs for veno-lymphatic insufficiency]. AB - Treatment of venous and lymphatic insufficiency of the lower limbs is based on 3 components: elastic support, venotonic drugs and radical treatments (surgery or sclerotherapy) of insufficient veins. Venotonic drugs have specific indications limited to functional impairment: heavy feeling in the legs, pain and impatience in the evening. There are different categories of venolymphatic drugs. Flavonoids have various pharmacological actions, most notably an increase in venous tone, reduction of capillary permeability and increase of capillary resistance. Choice of a venotonic drug is funded on knowledge of pharmacodynamics and pharmacokinetics of the molecule, critical evaluation of clinical studies, physician's personal experience and drug cost. Venotonic drugs are useful when venous insufficiency leads to functional manifestations. They are especially the treatment of heavy leg syndromes during warm seasons when elastic support is uncomfortable. PMID- 11008500 TI - [Manual lymphatic drainage]. AB - Manual physiotherapy of lymphoedema is the best treatment for chronic venous and lymphatic stasis of the lower limbs. It consists in handlings of the limb for the appeal and the resorption of the lymph but it cannot be used alone. It must be associated with other techniques: defibrosing, skin care, compressive bandages, muscular exercise. Contensive and compressive bandagings prolong the action of lymphatic drainage. The main indication of manual physiotherapy is primary or secondary lymphoedema but also chronic venous disease, and in some selected cases acute venous thrombosis and traumatic or post-traumatic pains. PMID- 11008501 TI - [Esthetic treatment of varicosities]. AB - Telangiectasia is dilatation of the subpapillary venous plexus of the epidermis of the lower limbs, which can lead to aesthetic embarrassment. Before treating telangiectasia, patient history and clinical examination help establishing its origin. It can be with isolated, associated reticular drainage veins, or be part of superficial venous insufficiency. Several types of treatment have been proposed. Microsclerotherapy is the most effective and least costly. Muller's phlebectomy can be performed when telangiectasia is fed by large reticular veins, either afferent or efferent. Treatment by laser and pulsed light appear best reserved to treatment of finer venous dilatations, either complementary or after failure of sclerotherapy. PMID- 11008502 TI - [Support and compression of the legs]. AB - Physical methods are essential in the treatment of venolymphatic insufficiency. The available materials have two modes of action, support and compression; they may be used alone or in association. Clinical experience and a few scientific works formed the basis for establishing protocols adapted to each situation. Like any major treatment, these should be under the full responsibility of physicians. Prescriptions should be precise and, in particular, evaluated for each patient by the physician himself. PMID- 11008503 TI - [Heat therapy in chronic venous insufficiency of the legs]. AB - Water cures have long been advocated for venous and lymphatic disorders. Spa therapy combines hydrotherapy, physical therapy and education. Immersion increases central blood volume, diuresis and natriuresis. These effects are independent of the mineral characteristics of the water. Hydrotherapy is viewed as a complementary treatment of venous and lymphatic insufficiency whatever its stage of development; but this has not been formally proved. PMID- 11008504 TI - [Perils in the Americas: five commentaries and an editorial]. PMID- 11008505 TI - [Exophthalmos. Diagnostic orientation]. PMID- 11008506 TI - [Hypertension in pregnancy. Diagnosis, complications, treatment]. PMID- 11008507 TI - [Chronic delusions. Diagnosis, development, treatment]. PMID- 11008508 TI - [Gait disorders and falls in the elderly. Diagnostic orientation]. PMID- 11008509 TI - [Infectious mononucleosis. Epidemiology, diagnosis, development]. PMID- 11008510 TI - [Disorders of acid-balance balance. Physiopathology, diagnosis, treatment]. PMID- 11008511 TI - [Drug utilization. Different sectors and determining factors]. PMID- 11008514 TI - [Bovine herpes mammillitis: clinical symptoms and serologic course]. AB - Bovine herpes mammillitis was diagnosed in a dairy herd with udder and teat skin lesions. Clinical symptoms seen in 6 cows consisted of round dry areas at the teats as well as large red and painful areas with crust formation at the teats, the teat basis and the udder. Diagnosis was verified by demonstrating numerous virus particles with the typical herpes structure and by BHV-2 serum neutralization test. Prevalence of BHV-2 in the herd was determined by using BHV 2 SNT at 7 occasions during a period of 15 months. The relatively low BHV-2 SNT titres as well as the seasonal increase of BHV-2 titres and seroprevalence in the month of September were indicative of a chronic and latent BHV-2 infection in the herd. PMID- 11008513 TI - [Ehrlichiosis in Switzerland--significance for veterinary medicine]. AB - Ehrlichiosis is a rickettsial disease of animals and humans caused by various species of Ehrlichia. These obligate intracellular microorganisms infect granulocytes or monocytes of mammalian hosts and are transmitted mostly by ticks. The febrile disease is often undiagnosed and characterized by leukopenia, anemia and thrombocytopenia. The diagnosis of ehrlichiosis is based on clinical findings, tick infestation, direct or indirect detection of the agent, and serology. The role of native and imported ehrlichiae of animals in Switzerland is discussed. PMID- 11008515 TI - [Prevention of pregnancy in bitches with the progesterone antagonist anglepristone (alizone)]. AB - A study was carried out between April 1997 and October 1998 to determine the effect of the progesterone antagonist, Aglepristone (Alizine), for the prevention of pregnancy. 93 bitches were treated, because of mismating, with Aglepristone. The owners were then contacted 2 weeks and 6 to 12 months after treatment to gather any information on effects noted over this period. The major questions were, was pregnancy prevented and if so what side effects were observed, if any, and whether any metropathies were diagnosed. Also noted was the beginning of the next heat and, if the bitch was mated, the fertility rate. Pregnancy was seen in only one bitch. In 51 bitches minor side effects, either singularly or in combination, such as a transient itch, vaginal discharge, reduced appetite, tiredness or attachment were observed. The fertility was not influenced by the treatment and the incidence of metropathies was unchanged. PMID- 11008516 TI - [Isolation of Listeria spp. and Aspergillus fumigatus--two case reports from mastitis diagnosis]. AB - Two rare cases of a bovine listeria mastitis and a mould mastitis are being described and discussed. The L. monocytogenes strains isolated from milk as well as silage samples were further genotyped by means of Pulsed-Field Gel Electrophoresis (PFGE) and macrorestriction analysis (ApaI). Three strains from the silage and four strains from quarter milking showed an identical PFGE pattern. PMID- 11008517 TI - [Torsio uteri as the differential diagnosis for gestational ketosis in sheep]. PMID- 11008518 TI - [Disintegration of the Nordic countries?]. PMID- 11008519 TI - [The Nordic countries--closest cooperators when it comes to research?]. PMID- 11008520 TI - [Social medicine should bite the dust!]. PMID- 11008521 TI - [A difficult problem without an easy solution]. PMID- 11008522 TI - [Tattooing--a cultural expression]. PMID- 11008523 TI - [Ophthalmology and cosmetics]. PMID- 11008524 TI - [A potential for prevention of bicycling-related head injuries]. AB - BACKGROUND: Bicycle helmets prevent head injury in bicycle riders. Still, only a portion of bicycle riders in Norway use bicycle helmets. The aim of this study was to estimate the number of head injuries among bicycle riders that might be prevented by increased helmet use in Norway. MATERIAL AND METHODS: We used data from the Norwegian National Injury Register for the years 1995 and 1996 to estimate the number of bicycle injuries in Norway. In order to estimate the number of bicycle users and helmet users in different age groups, we used data from earlier surveys of bicycle use. Data on the effectiveness of helmet promotion interventions have been obtained from a systematic review of the literature. RESULTS: The overall annual incidence rate was 92 injuries per 100,000 bicycle users. The incidence varied with age and was highest among children. If every rider used a helmet, about 1,600 head injuries would be avoided every year, of these, 800 among children aged 0-14. Currently available helmet promotion interventions may improve the use among children by about 40%, thus preventing about 1,500 head injuries over a period of three years. Successful helmet promotion interventions use a combination of health education and helmet distribution strategies. INTERPRETATIONS: There is a significant health improvement potential in promoting bicycle helmets in Norway by implementation of evaluated modes of intervention. PMID- 11008526 TI - [Centrally acting muscle relaxants and traffic hazards]. AB - BACKGROUND: An increasing number of the centrally acting muscle relaxants were withdrawn from the Norwegian market during the 1988-98 period. The only drug in this group now marketed in Norway is carisoprodol. The National Institute of Forensic Toxicology in Norway analyses all blood samples from suspected drugged drivers. In later years there has been a marked increase in the number of blood samples testing positive for carisoprodol or meprobamate (the major metabolite). MATERIAL AND METHODS: 480 cases testing positive for central muscle relaxants in the years 1984-1998 were further studied. RESULTS: Compared with blood samples positive primarily for benzodiazepines, there were more women in the group (39% vs. 15%), and fewer drugs and less alcohol were detected. INTERPRETATION: The positive samples may indicate misuse or abuse due to the fact that high drug concentrations and concomitant use of benzodiazepines were frequent. This knowledge should have implications for doctors prescribing centrally acting muscle relaxants. PMID- 11008525 TI - [Treatment of patients with acute head injury in Vestfold 1987-96]. AB - BACKGROUND: In Norway, patients with severe head injuries are transported to a regional, neurosurgical department for surgery, but some are operated on by surgeons without neurosurgical training in local hospitals. MATERIAL AND METHODS: Data were retrospectively collected from the records of all patients (n = 161) hospitalised alive with a severe head injury occurring within Vestfold County (1987-96). RESULTS: Overall lethality was 27%. 54 patients (34%) underwent decompressive surgery. 31 patients were operated in the local hospital, by 13 different surgeons. 30 of these patients had extracerebral haematomas. These patients had a significantly worse outcome than the 23 patients operated in a regional neurosurgical department. Patients with an epidural haematoma had a better outcome than patients with an acute subdural haematoma. We retrospectively classified ten operations in the local hospital as inadequate. INTERPRETATION: In Norway and countries with a similar hospital system, local hospitals should establish guidelines for safe and swift transport of head injury patients to the nearest neurosurgical department, and should not try to perform neurosurgical decompression in such patients. PMID- 11008527 TI - [Evaluation by public health physicians of collaboration and workload within municipal health services]. AB - BACKGROUND: We wanted to explore the quality and importance of the collaboration between public health physicians and selected groups of professionals and politicians. MATERIAL AND METHODS: The study was a cross-sectional survey based on self-administered questionnaires with closed questions. Response came from 218 public health doctors who worked in the municipalities throughout the study period (1988-91), and from 98 doctors who left public health during the same period (81% and 86% of the study populations respectively). RESULTS: The doctors reported good and important collaboration with other health professionals, but less good and less important collaboration with other professionals and with politicians. With increased administrative distance between the doctor and other municipality staff, the perceived quality of the collaboration decreased. INTERPRETATION: Closer collaboration between public health doctors and important decision makers in the municipality promotes job satisfaction among doctors and may reduce turn-over. PMID- 11008528 TI - [Tryggve Gran--the first Norwegian heroic pilot]. AB - Tryggve Gran grew up in an affluent family in Bergen, Norway. The German emperor, William II, often visited the families of his friends. Gran became a good skier, hence well prepared for Robert Scott's second expedition to the Antartic in 1910. Gran deeply regretted the Scott-Amundsen competition, and was cut off from the team heading for the South Pole. In 1913, Gran trained in England and France as an air pilot. On 30 July 1914 he became the first pilot to cross the North Sea from Scotland to Norway. He joined the Royal Air Force in 1916 under the pseudonym of Teddy Grant, passing himself off as a Canadian, and received the Military Cross for distinguished war service. During the Second World War, Gran was a member of Quisling's pro-German National Party. A commemorative stamp was issued in 1944 on the 30th anniversary of his North Sea flight, and a meeting held in his honour with Quisling and German officers present. In this article, the author discusses some psychological aspects of Tryggve Gran's choice of tasks and of his politics. Gran lost his father when he was only five and when he was 11 he was sent off to a pension in Switzerland for a year. Strongly ambivalent feelings from the oedipal period and from the latency may later have been released through hazardous activities, certainly with self-destructive aspects. His membership in Quisling's party might be seen in this context. PMID- 11008529 TI - [Sound and unsound around the Sound. A congress report on health differences between Denmark and Sweden]. AB - During the past 10 years, researchers on both sides of The Sound in the so-called Oresund region have worked together to analyze the causes of the observed differences in life expectancy between Denmark and Sweden. The region includes Copenhagen and North Zealand and the county of Scania in southern Sweden, with Malmo as its largest city. Both Denmark and Sweden held top rankings among OECD countries in 1970 regarding life expectancy at birth. In 1990 Denmark had fallen to a bottom ranking, while Sweden was still at the top. At a scientific meeting in Malmo on January 26-27, 2000, some 60 presentations were made concerning differences and similarities in health measures on the two sides of the sound which divides the two countries. On the Swedish side, life expectancy is 3-4 years longer than on the Danish side. Paramount among explanations are higher mortality figures due to smoking and alcohol-related diseases on the Danish side, both in men and women (most pronounced). Minor contributions to the differences are suicides and traffic accidents. Historically, Denmark and Sweden have much in common. Until 1658, Scania, the Swedish county to the east of The Sound was part of Denmark. During the past 150 years rather impressive cultural differences have developed. Smoking prevalence and alcohol consumption are more than twice as high on the Danish side of the sound. In coming years, researchers, representatives for the health service systems and others will work together in order to prevent disease and promote health in the Oresund region. The conference was arranged by the Medicon Valley Academy, an EU-supported enterprise seeking to stimulate research and development within the health sector in the Oresund region (also known as Medicon Valley). PMID- 11008530 TI - [Scandinavian guidelines for management of minimal, mild and moderate head injuries]. AB - The Scandinavian Neurotrauma Committee (SNC) was initiated by the Scandinavian Neurosurgical Society to develop evidence-based guidelines for improved care of neurotrauma patients. A MEDLINE search identified 475 papers dealing with the management of minimal, mild and moderate head injuries. Fourty-two studies presenting Class II evidence on the initial management of such injuries were reviewed and management guidelines were developed. Implementation of the Head Injury Severity Scale is advocated. Patients with minimal injuries (no loss of consciousness (LOC), Glasgow Coma Scale (GCS) score 15) can be safely discharged. Routine early CT scan is recommended in cases with mild injuries (history of LOC, GCS 14-15) and patients with normal scans may be discharged. CT scan and admission is mandatory in moderate injuries (GCS < or = 13). All patients harbouring additional risk factors should be scanned and admitted. A flow chart for clinical decision making and a Head Injury Instruction card are introduced. PMID- 11008531 TI - [Norwegian tourists' use of medical services abroad]. AB - BACKGROUND: Previous studies have shown that one in three holidaymakers experienced illness on their trip abroad. We wanted to find out what happens to Norwegian tourists. MATERIAL AND METHODS: We have collected data from the Association of Norwegian Tour/Operators on travel patterns, and from a travel insurance company on illness and expenses. RESULTS: Between 1.5% and 5% of travellers get so ill that they need medical assistance. The number of consultations that results in hospitalisation vary from less than 5% in one country to more than 30% in neighbouring territories. Average cost of hospitalisation differ considerably from one country to another. INTERPRETATION: It appears that cultural differences, differing treatment facilities, and varying required rates of return are decisive for whether or not a patient is referred to hospital. PMID- 11008532 TI - [Long term effects o gastric banding for weight reduction]. AB - BACKGROUND: Obesity is defined as body weight exceeding normal weight by 30%, or a body mass index (BMI) > 30. Various surgical procedures have been introduced to treat obesity. Gastric banding is a variant of the gastroplastic method. Our aim was to study the long term effect of this procedure. MATERIAL AND METHODS: Over the years 1982 to 1988, gastric banding was performed at the county hospital of Haugesund in 26 patients with a mean age of 35.7 years and a mean preoperative BMI of 41.2. Data were obtained from patient files and from questionnaires returned by 24 patients, on average 14 years after the operation. RESULTS: Mean BMI for 16 patients with an intact banding device at follow-up was 35.2. Mean weight reduction was 15 kilograms. 58% of the patients regretted the procedure. Only a few were completely satisfied. INTERPRETATION: Long term results of gastric banding are not satisfactory. PMID- 11008533 TI - [The obesity epidemics--do diet pills have a place in the treatment?]. AB - BACKGROUND: Many experts consider obesity a chronic disease that may require long term therapy. A loss of 5-15% of body weight is associated with improvements in cardiovascular risk factors and morbidity. However, most studies show that the majority of patients who lose weight relapse. Patients may be unable to maintain a low energy intake when confronted with an almost limitless supply of food. Moreover, a number of physiological mechanisms favour a set point for body weight, that may be altered with anti-obesity drugs. MATERIAL AND RESULTS: In the current paper we describe actions and effects of current anti-obesity drugs. The centrally acting drug, sibutramine, is an adrenaline and serotonine re-uptake inhibitor which was recently approved in the USA for obesity. The USA, the European Union and Norway have approved orlistat, a pancreatic lipase inhibitor for weight reduction for up to two years. Patients must maintain a low fat intake in order to avoid gastrointestinal discomfort. In recent studies, orlistat and diet reduced body weight by 9% versus 6% on placebo and diet. No studies have documented long-term safety of anti-obesity drugs. INTERPRETATION: Treatment of a lifestyle-related disease like obesity with medications is controversial, however, such treatment may not differ substantially from treatment of type II diabetes, hyperlipidaemia or hypertension. PMID- 11008534 TI - [Is the benefit of organized mass screening for cervix cancer and breast cancer in Norway scientifically justified?]. AB - BACKGROUND: It has been argued that mass screening may reduce mortality from cervical cancer and breast cancer in Norway by 50% and 39%, respectively. However, some authors doubt that mass screening for breast cancer is justifiable. This article discusses whether mass screening for cervical cancer in Norway is more effective than opportunistic screening. MATERIAL AND METHODS: The scientific methods used to justify organised mass screening for cervical cancer are reviewed and discussed, as are the randomised trials used to justify screening by mammography. The author is especially critical of the use of descriptive statistics to justify screening, on the assumption that there is a basic constant incidence or mortality rate. RESULTS: The incidence rates of pre-malignant conditions and cervical cancer in Norway are presented. There is no essential increase in the number of pre-malignant conditions after three years of organised mass screening compared to the previous three-year of unorganized screening. However, the number of pre-malignant conditions did increase from 1987 to 1992 due to a national recommendation for testing every three years. By contrast, incidence rates for cervical cancer remained almost constant in the 1987-97 period. INTERPRETATION: There is no scientific proof of organised mass screening for cervical cancer being preferable to unorganized screening. The author warns against only comparing observed rates with historic rates and rates in other countries; alternatively, rates among those who participate in the screening might be compared in a simultaneous analysis to rates among those who do not respond to the invitation. The author also suggests estimating correlation coefficients between individual test results to identify high-risk individuals. PMID- 11008536 TI - [Painful neuropathies]. PMID- 11008535 TI - [Subacute abdominal pain in a very old woman]. PMID- 11008537 TI - [Gastric lavage and activated charcoal in acute poisonings]. PMID- 11008538 TI - [What is specifically Nordic in medicine?]. AB - Nordic health professionals often hear the term "the Nordic model" when visiting countries outside the Nordic area. This Nordic model is usually related to selected aspects like the doctor-patient-relationship, the structures and functions of the Nordic health systems, and the independent, nationwide control systems in the Nordic countries which deal with research ethics and integrity in research within the biomedical sciences. In this essay the concept of specifically Nordic characteristics within medicine and health services is evaluated through an observation window. The conclusion is that the most components of the Nordic health sectors, including education and research, are by their very nature global, but that their weighting and the projection to clinical work and research show decisive traits, especially related to patients' rights and ethics. On this background, it seems legitimate to accept the surrounding world's impression of a special Nordic model. PMID- 11008539 TI - [Differences in health status--policy, life style and genes]. AB - BACKGROUND: The five Nordic countries seem rather homogeneous when assessed from abroad. They do have a lot in common: similar political systems, same standard of living, and even a common language root for a large part of the population. The Nordic countries have collaborated quite closely after the Second World War in fields like the labour market, pension rights and passport control. However, the health status has varied considerably during the twentieth century, even though death rates from some of the most common diseases such as cardiovascular disease are starting to converge. MATERIAL AND METHODS: Vital statistics have been compared with regard to some common diseases. The differences are described according to current knowledge on preventive programmes such as breast cancer screening and screening for cardiovascular risk. The prevalence of some lifestyle variables such as alcohol, smoking, physical activity and fat consumption are reported. RESULTS AND INTERPRETATION: Lung cancer and cancer of the stomach still show substantial variation between the countries. Both discrepancies and similarities must be related to lifestyle, culture, politics and to some extent genetics. The five countries have their own viewpoints on tobacco, alcohol and nutrition policy, with the Danes the more liberal, and Swedes, Finns and Norwegians pursuing a stricter policy. These differences are to some extent reflected in the incidence of lung cancer, which are more positive in Finland, and the high death rates from liver cirrhosis in Denmark. The relatively unsatisfactory development in Denmark may be a consequence of a much longer period of structural unemployment than the other countries, as well as a more liberal attitude towards alcohol and smoking. PMID- 11008540 TI - [Health care systems in the Nordic countries--more similarities than differences?]. AB - The aim of this paper is to explore similarities and differences of the Nordic health care systems. The analysis is based on a three-part model involving patients, providers and the financing third party. In all Nordic countries, about 80% of the funding come from public sources. In Iceland, central government is providing most of the health care services, while the county councils are central in Denmark, Norway and Sweden. In Finland, the municipalities are providing most of the health care. Salary is the only payment system for general practitioners in Iceland, while it is used for some GPs in Finland, Norway and Sweden. Capitation in combination with service fees is used for all Danish GPs and the majority of the Finnish, while various fee-for-service systems are used for the others. All Nordic countries had global hospital budgeting in the 1980s; since then, Finland, Norway and Sweden have implemented others systems, predominantly combinations of diagnosis-related group financing and global budgets. The amounts of resources devoted to health care are about the same in all five countries whether measured by the proportion of GDP devoted to health care, or by hospital beds or doctor/patient ratios. In monetary terms, Denmark, Iceland and Norway spend more than Finland and Sweden. Despite similar amounts of resources, they are quite differently used across the countries. Differences of a factor of two or more are observed for hip operations, gal bladder operations and pharmaceuticals. Danes and Finns are very satisfied with their health care system; the Swedes are not. All the Nordic countries have increased patient co payments during the 1990s. Finnish patients may pay an extra free to get a one bed room. Finland also has the greatest number of private hospitals. Despite all these differences, the Nordic health care systems are quite similar when seen in a global perspective. PMID- 11008541 TI - [The manpower market for physicians in the Nordic countries 1980-2000]. AB - The ratio between population and active physicians in the Nordic countries has improved from 488 inhabitants per physician in 1980 to 315 in 2000. There is a large mobility of physicians between the countries, contributing to levelling out swings in demand and supply of manpower. Language and culture being similar, physicians can easily adjust to working in a neighbouring country. Iceland is special in this respect, as a surplus of Icelandic physicians has always found work in the other Nordic countries. Of course, their numbers are small relative to the total number of active physicians in the Nordic countries, now approximately 76,000. The number of students admitted to Nordic medical faculties has changed in line with swings in estimated future demand for manpower. Today, numbers are increasing again; this year, approximately 2,900 students will be admitted. Norway stands apart from the other Nordic countries in terms of medical manpower needs. During the last 20 years there has been a continuous shortage of physicians while all the other countries have been through periods of surplus and unemployment among physicians. Manpower forecasts in the early 1980s underestimated the growth in the health care system and hence the demand for medical manpower. PMID- 11008542 TI - [Continuing education in the Nordic countries]. AB - The Nordic Labour Market Agreement gives Nordic medical specialist the right to be certified in the other Nordic countries. The agreement also provides for cooperation on specialist training between the five countries. This article is mainly based on published statements from the joint working group for medical manpower forecasts and specialist training in the Nordic countries. There are great differences between the countries in the way specialist training is organised. An underlying assumption in the Nordic agreement is that the level and quality of specialist training is the same in all five countries; however, there does not seem to be a corresponding coordination between the training systems adopted. PMID- 11008543 TI - [Surgery of strabismus--"cosmetic indication"?]. PMID- 11008544 TI - [Is carisoprodol justified as a muscle relaxant?]. PMID- 11008545 TI - [Physicians and alcohol--differences between the Nordic countries?]. PMID- 11008546 TI - [Whistleblowers--are they "difficult persons"--or are they forced to be difficult?]. PMID- 11008547 TI - [Hospital ship--it's there when needed]. PMID- 11008548 TI - [Greetings from the Mecca of epidemiology]. PMID- 11008549 TI - [Norwegian health services as seen from Stockholm]. PMID- 11008550 TI - [Ultrasonography early in pregnancy]. PMID- 11008551 TI - [The National Board of Health and Welfare recommends an inefficient method]. PMID- 11008552 TI - [X-Ray upper airway changes in individuals suffering from obstructive respiratory disorders during sleep]. AB - Obstructive respiratory disorders during sleep present an important medical and social problem. Serious dysfunctions of cardiovascular, nervous, endocrine and other vital systems of the body reduce longevity and life quality. On the other hand, load nocturnal snore and abnormal diurnal sleepiness cause great damage to family life, reduce working capacity and induce accidents. X-ray visualization of the upper airways is essential in diagnosing obstructive upper airway states and selecting patients for surgical treatment. The paper presents the authors' own experience in using various X-ray diagnostic methods in patients with chronic snore and obstructive sleep apnea-hypopnea syndrome. PMID- 11008553 TI - [Radiation diagnosis of Langerhans cell histiocytosis (LCH) of lung]. AB - The authors studied the pattern of X-ray changes detectable in the lungs of 41 patients with LCH. The results of classical X-ray study were supplemented by the data of computed tomography, ultrasonography, and 99mTc and 67Ga citrate radioisotopic studies. The diagnostic values of these techniques were compared. X ray syndromes of changes in the lung in LCH were found to be interstitial, interstitial and granulomatous, tumorous (focal). The authors emphasize the multisystematic character of a lesion, the phasic nature and progression of the disease into fibrosis. PMID- 11008554 TI - [Chronic occlusions in patients with coronary heart disease: results of recanalization]. AB - The paper analyzes the results of endovascular treatment in 298 patients with chronic occlusions or subtotal stenoses of coronary arteries. The authors show that balloon coronary angioplasty for chronic coronary occlusions is an effective and relatively safe techniques of myocardial revascularization. Successful endovascularization has been achieved in 68.1% of patients with chronic coronary occlusions and in 97.5% of those with subtotal coronary stenoses. The results show a high incidence of restenosis and asymptomatic reocclusion. PMID- 11008555 TI - [Use of radial artery as an access during coronary angiography: experience and further prospects]. AB - The authors present their own experience in using the radial artery for diagnostic coronarography. Twenty two transradial studies were made during a year and a half. Following the studies, complications were as follows: failure at puncture in 1 (4.5%) patient, local and cardial complications being absent. The authors concluded that the radial access is a promising alternative to the routine femoral one now. The use of the radial access opens up new vistas for coronary angiography performed in the outpatient setting, by making this technique accessible to patients with coronary heart disease who need to be thoroughly studied. PMID- 11008556 TI - [Interventional radiology in congenital and acquired cardiovascular diseases]. AB - The paper outlines the present-day achievements in interventional radiology in cardiovascular diseases: balloon valvuloplasty in cardiac diseases (isolated pulmonary arterial stenosis, aortic and mitral stenosis), balloon vasodilatation (peripheral pulmonary arterial stenosis, aortic coarctation), embolization of the vessels and pathological communications, atrioseptostomy, transcatheter closure of atrial septal defects. PMID- 11008557 TI - [Current approach to treating borderline coronary stenosis: clinical case]. PMID- 11008558 TI - [Up-to-date positions of prophylactic fluorographic examinations of chest (review of literature)]. PMID- 11008559 TI - [Radiation diagnosis of biliary system before laparoscopic cholecystectomy. Part II: non-ultrasonic techniques]. PMID- 11008560 TI - ["Diamag" is a new magnetic resonance tomograph ("Az" firm)]. PMID- 11008561 TI - [To applicability of foreign medical radiographic films during various chemophotographic treatments]. PMID- 11008562 TI - [Recovery processes in the cerebral cortex, myocardium and thymus of rats with experimental atherosclerosis exposed to low-frequency electromagnetic fields on the head]. AB - Studies of animals with experimental sclerosis has shown that a course of 10 procedures of alternative magnetic field (AMF) (50 Hz, 30 mT, 3 min daily) promotes partial recovery of the lipid spectrum and corrects vasomotor-metabolic disturbances in the cerebral cortex, myocardium and thymus caused by atherosclerosis. Combination of AMF with constant magnetic field in the same regime and location does not produce a hypolipidemic effect in atherosclerotic animals and this, in combination with increased vascular permeability may aggravate the condition. Activated microcirculation, antioxidant and antiproteinase effects in activation of biosynthetic processes in the cerebral cortex reflect inhibition in the CNS in this combined effect and create conditions for a hypotensive effect. PMID- 11008564 TI - [The combined effect of potable mineral waters and mid-alpine factors on hormonal status (clinico-experimental research)]. AB - Hormonal status was examined in 30 intact male Wistar rats (body mass 200-220 g) and 10 healthy male volunteers aged 25-35 years. Hormonal status and influence of exercise were assessed in human males after 2, 10 and 19 days of stay in the sanatorium situated 2000 m above the sea level. The animals were decapitated on day 24 of their stay in the mountains. Blood from the rats' cervical vessels was examined for glucose, beta-lipoproteins, triiodothyronine, thyroxine, TTH, hydrocortisone. The findings indicate that stay in conditions of medium-height mountains climate aroused adaptation reactions in the hormonal systems, i.e. activation of thyroid hormones. An additional course of mineral water contributed to further rise of the blood hormones. Hormonal trends in the volunteers were similar. There was also a rise in the levels of beta-lipoproteins which was arrested by physical exercise. PMID- 11008563 TI - [Pain relief by low-intensity frequency-modulated millimeter waves acting on the acupuncture points]. AB - Analgetic effect of low-intensive frequency-modulated millimetric waves (MW) was studied in mice with formalin induced nociceptive behavior reaction (licking of defeat hindpaw). MW were applied to the acupoint E 36 of the defeat hindpaw. The following MW were used: 60 GHz (1) and 118 GHz (2) which were modulated by 4 Hz; noise MW within the range of 42-95 GHz (3) and 90-140 GHz (4) which were modulated in accidental order by frequencies 1-60 Hz; combinations of fixed frequencies with noise - 60 GHz + noise 42-95 GHz (5) and 118 GHz + noise 90-140 GHz (6). All used MW combinations suppressed licking of the defeat hindpaw and increased duration of sleep and eating. The strongest analgesia was achieved in series 1-3 (42.4-69.7%), the weakest in series 6 and 4 of the experiment (12.2 19.7%). PMID- 11008565 TI - [An evaluation of the effect of sodium bicarbonate-chloride mineral water on the rat immune system]. AB - Mineral water Uralochka tried in rats is obtained from the spring in the territory of the sanatorium Ural (Chelyabinsk Province). The mineral water contains chloride, hydrocarbonate ions and sodium cations, mineralization is 4.13 g/dm3. A 15-day course of Uralochka drinking did not influence much reaction of delayed hypersensitivity, but increased the absolute number of antibody-forming cells by 95.86%, raised the ability of peritoneal macrophages to absorb latex particles. Thus, mineral water Uralochka can raise intensity of humoral immune response and the ability of macrophages for phagocytosis of the latex particles. Phagocytosis activity and intensity increased by 48.27 and 62.13%, respectively. PMID- 11008566 TI - [Carbonate mineral baths in the combined treatment at a piedmont health resort of patients operated on for rheumatic heart defects]. AB - The response of the cardiovascular system of 64 patients operated for rheumatic heart disease to mineral carbon dioxide baths was studied in the course of rehabilitation in low-mountain health resort. It was found that the above baths have a training effect on the cardiac muscle, improve myocardial metabolism. They are an essential factor in combined sanatorium treatment of patients after surgical correction of valvular defects. Baths-induced changes in the central and peripheral hemodynamics depend on cardiovascular function. Therefore, they require an individual approach. If the exposition increases to 15 minutes, mineral carbon dioxide baths have more potent training effect on the myocardium and raise effectiveness of the sanatorium treatment. PMID- 11008567 TI - [The efficacy of combined treatment in patients with chronic mycotic bronchitis using baro-laser therapy]. PMID- 11008568 TI - [The methodological aspects of using cryomassage on patients operated on for discogenic neuropathies]. AB - Cryomassage in combined treatment of 100 patients operated for discogenic neuropathy produced a pronounced vasotropic effect. The authors have determined optimal zones for cryomassage application and order of its usage with some physical factors. PMID- 11008569 TI - [The use of physical factors in the rehabilitative treatment of patients with stone fragments in the upper urinary tract after extracorporeal shockwave lithotripsy]. AB - 155 patients with fragments of the crushed stones in the upper urinary tract after lithotripsy were divided into 3 groups. 60 patients of group 1 received dynamic amplipulse therapy, oral mineral water, iodobromine baths. Elimination of the fragments was observed in 91.7% of the patients. Group 2 patients (n = 60) were exposed to local vibration, ultrasound and iodobromine baths. The effect occurred in 96.7% of the cases. 35 patients of group 3 were exposed to impulse low-frequency magnetic field and took iodobromine baths. The elimination of the fragments occurred in 62.9% of the cases after a single treatment course. PMID- 11008570 TI - [The combined use of pelotherapy, ultrasound and sinusoidal modulated currents in chronic prostatitis and proctosigmoiditis]. PMID- 11008571 TI - [A comparative assessment of different cryotherapy methods for patients with chronic nonspecific salpingo=oophoritis]. AB - 97 females at reproductive age with chronic nonspecific salpingo-oophoritis (CNSO) were examined and treated. The results of the treatment (vaginal and external impact) demonstrate positive effects of various cryotherapeutic techniques on CNSO clinical course, on hormonal and immune unbalance, functional activity of the uterine tubes, regional hemodynamics, psychoemotional status. Thus, cryotherapy is an effective adjuvant in combined therapy of CNSO. PMID- 11008572 TI - [An experimental and clinical validation of a method for treating acute inflammatory diseases of the adnexa uteri by using the electrophoresis of a therapeutic mud preparation]. AB - Experimental and clinical investigations have demonstrated that the addition of abdominal-sacral electrophoresis of esobel 1% solution to combined therapy of acute inflammation of the uterine appendages improves treatment results. Bioantioxidants bring about the antiinflammatory action of the mud preparation esobel while its high-polar lipids prevent structural-metabolic impairment of the appendages' tissue elements as shown by a pronounced antiexudative effects and weak secondary alteration. PMID- 11008573 TI - [The new potentials of magneto-laser therapy and electrostimulation in children with cleft palate and upper lip]. AB - The paper presents rationale for design of new procedures of physiotherapy in children with expanded palate and upper lip; morphofunctional features of prenatal and early postnatal formation of central nervous system; a multilayer technique of magneto-laser radiation to the projection of the anterior central gyrus and the technique of electrostimulation by the system of the wink reflex which can influence central mechanisms of regulation and adaptation before and early after uranoplasty and cheiloplasty. PMID- 11008574 TI - [The sauna in the treatment of children with atopic dermatitis]. AB - Complex sauna therapy for children with atopic dermatitis gives positive clinical results and improves the condition of their autonomic nervous system. Marked clinical response but unsatisfied autonomic nervous system regulation necessitate long-term sauna treatment to cure this disease. PMID- 11008575 TI - [The health resort and recreational potential of Moscow Province]. PMID- 11008576 TI - [The regulation of the sanatorium-health resort market]. PMID- 11008577 TI - [Endonasal electrophoresis in ophthalmology]. PMID- 11008578 TI - [The biorhythmological regulation of external breathing in bronchial asthma patients by using intermittent normobaric hypoxia therapy]. PMID- 11008579 TI - [The treatment of ENT diseases in children of the indigenous nationalities of the North with local mineral waters from Zavodoukovski spring]. PMID- 11008580 TI - [The use of hydrocortisone combined with ultrasound with gonarthrosis patients]. PMID- 11008581 TI - [A patent review on the problem of the transfer of the informational properties of therapeutic factors]. PMID- 11008582 TI - [The effect of physical factors on the skin permeability of normal white rats and in the modelling of pathological processes]. PMID- 11008583 TI - African trypanosomiasis, tuberculosis and dengue research. PMID- 11008584 TI - American Society of Critical Care Anesthesiologists 13th annual meeting. San Francisco, California, USA. October 13, 2000. Abstracts. PMID- 11008585 TI - Psychotropic drugs and preschoolers. PMID- 11008586 TI - Ischaemic heart disease: across the social and geographic divides. PMID- 11008587 TI - SIDS: facts and controversies. PMID- 11008588 TI - Falls in the elderly: what can be done? PMID- 11008589 TI - Characteristics and outcomes of older patients presenting to the emergency department after a fall: a retrospective analysis. AB - OBJECTIVES: To study older patients presenting to the emergency department after a fall--factors associated with the fall, injuries sustained and outcome. DESIGN: A retrospective analysis using the Emergency Department Information System (EDIS), the Trauma Registry and the patient information database (CCIS), in addition to the patient's emergency and inpatient medical records. SETTING: Emergency department of a major inner city teaching hospital, 1 June-30 November 1997. PATIENTS: All patients over 65 years presenting to the emergency department (ED) after a fall, for whom complete medical records were available. RESULTS: Of 803 patients over 65 years presenting to the ED after a fall, complete records were available for 733 (91.3%) (283 men and 450 women). Extrinsic (accidental) causes were implicated in more than a third of falls (313 patients [42.7%]). A high proportion of the patients were living at home (520; 70.9%) and walking unaided (389; 53.1%). Although absolute numbers of women increased with age, men were as likely as women to present after a fall. Many patients had fallen before- 39% of the men (111/283) and 24% of the women (110/450). In 78 patients (10.6%), alcohol misuse may have been a direct cause of the fall. The overall injury rate was 70.5% (517/733 patients), the most common injury being an isolated fracture (269/517 patients; 52.0%). In all, 419 patients (57.2%) were admitted to hospital, 48% (200/419) with a fracture and 52% (219/419) for investigation of the medical cause of the fall. The median length of hospital stay was 6 days (mean, 10.4 days; range, 1-129 days); 35% (146/419) of patients were in hospital for more than 10 days. CONCLUSION: Older patients presenting to the ED after a fall had high injury rates, high admission rates and often prolonged hospitalisation. About a third had fallen before. Patients at risk can be identified in the ED and referred to falls prevention programs. PMID- 11008590 TI - Predictors of bone mass in healthy older men in the community. AB - OBJECTIVES: To determine the predictors of bone mass (in lumbar spine and femoral neck) in healthy older men living in the community. DESIGN: Cross-sectional study. Bone mineral density (BMD) was measured and known predictors of bone mass and bone turnover were assessed. SUBJECTS AND SETTING: 113 independent, healthy men (70 years and over), not taking glucocorticoid therapy and without medical conditions known to affect bone mass and bone turnover, were recruited from recreational and sports clubs in southern Sydney in April/May 1997. MAIN OUTCOME MEASURES: BMD (measured by dual-energy x-ray absorptiometry); known predictors of bone mass (height; weight; body mass index; calcium level; serum 25 hydroxyvitamin D and free testosterone levels); and markers of bone turnover (serum bone Gla protein and procollagen-1 concentrations, and urinary deoxypyridinoline excretion rates). RESULTS: The mean age of the men was 76.6 years (range, 70-92 years). Mean (SE) BMD of the lumbar spine was 1.143 (0.019) g/cm2 and that of the femoral neck was 0.897 (0.013) g/cm2. BMD values indicating osteoporosis were found in the lumbar spine in 13 men (11.5%) and in the femoral neck in 35 men (31%). The best predictor of lumbar spine BMD was weight (R = 0.37; P = 0.001), and weight- and age-predicted femoral-neck BMD (R = 0.49; P < 0.001). The study group was analysed in two groups with BMD higher or lower than median bone mass corrected for age. Men with lower femoral-neck BMD for age had significantly lower weight, lower lean mass and higher bone Gla protein concentrations. In addition, men with lower lumbar spine BMD for age also had significantly lower fat mass. CONCLUSIONS: These data indicate that measures of body composition, such as weight and lean mass, are the main predictors of bone mass in healthy, community-dwelling older men. PMID- 11008591 TI - Prevalence of Huntington disease in New South Wales in 1996. AB - OBJECTIVE: To estimate the prevalence of Huntington disease (HD) in New South Wales on Australian Census Day (6 August) 1996. DESIGN: Survey of records of the Huntington Disease Service and major hospitals, and of neurologists, psychiatrists, clinical geneticists and genetic counsellors. SUBJECTS AND SETTING: All patients in NSW who, on Census Day 1996, either had a definite diagnosis of HD (motor signs of chorea or ataxia and family history of HD or positive DNA test result) or would have had signs and later received a definite diagnosis (assessed 1 April 1997 to 1 July 1999). MAIN OUTCOME MEASURES: Prevalence (HD patients per 100,000 population); patient characteristics; year and basis of diagnosis. RESULTS: 380 patients with definite HD were identified, giving a prevalence of HD in NSW in 1996 of 6.29 per 100,000 population (95% CI, 5.68-6.96). A third of HD patients were aged 60 years or older. Diagnosis was confirmed by DNA testing for 171 patients (45%), including 30 (8%) with no recorded family history. Average numbers of new diagnoses per year were 11.8 (1984-1988), 21.8 (1989-1993) and 28.6 (1994-1998). Estimated number of people with a 50% risk of inheriting the HD mutation was 25.2 per 100,000 population. Estimated incidence of HD in 1996 was 0.65 per 100,000 population. CONCLUSIONS: Prevalence of HD in NSW is similar to estimated prevalence in other Australian and Western populations. Increasing numbers of cases are being diagnosed, and the 18 chronic care beds currently designated for HD patients in NSW are unlikely to be sufficient. PMID- 11008592 TI - Driveway motor vehicle injuries in children. AB - OBJECTIVES: To describe the frequency, nature and outcome of driveway injuries in children. DESIGN: Retrospective case series of driveway-related injuries in children under 16 years of age admitted to the New Children's Hospital (NCH), New South Wales, from November 1995 to February 2000, and deaths reported to the New South Wales Paediatric Trauma Death (NPTD) Registry from January 1988 to December 1999. MAIN OUTCOME MEASURES: Circumstances of injury; type and number of injuries identified. RESULTS: 42 children were admitted to our institution with driveway related injuries over four years and four months. These represent 12% of all children admitted with pedestrian motor vehicle injuries. Fourteen deaths (including one of the children admitted to NCH) were reported to the NPTD Registry over 12 years, accounting for 8% of all paediatric pedestrian motor vehicle deaths reported to the registry. Typically, the injury involved a parent or relative reversing a motor vehicle in the home driveway over a toddler or preschool-age child in the late afternoon or early evening. Four-wheel-drive or light commercial vehicles were involved in 42% of all injuries, although they accounted for just 30.4% of registered vehicles in NSW. These vehicles were associated with a 2.5-times increased risk of fatality. In 13 of the 14 deaths, the cause was a severe head injury not amenable to medical intervention. CONCLUSIONS: Driveway injuries in children account for a significant proportion of paediatric pedestrian motor vehicle injuries and deaths in NSW. Prevention represents the only effective approach to reducing deaths from this cause. PMID- 11008593 TI - Wasp sting mortality in Australia. AB - Wasp sting fatalities have rarely been reported in Australia. We used data from the Australian Bureau of Statistics and State coronial authorities to investigate deaths from wasp stings in Australia from 1979 through 1998. Seven cases were identified, all involving men in rural settings. Five of the seven victims had prior histories of wasp or bee venom allergy, or both, but none carried injectable adrenalin. All patients with a history of systemic Hymenoptera sting allergy should undergo assessment for immunotherapy and carry adrenalin. PMID- 11008594 TI - Delay times and management of acute myocardial infarction in indigenous and non indigenous people in the Northern Territory. AB - OBJECTIVES: To investigate differences in presentation and management of Indigenous and non-Indigenous patients hospitalised with acute myocardial infarction (AMI). DESIGN: Retrospective review of hospital medical records. PARTICIPANTS AND SETTING: 122 patients with definite or possible AMI admitted to hospitals in the Top End of the Northern Territory (NT) in 1996. MAIN OUTCOME MEASURES: Percentage receiving thrombolytic therapy; delays from symptom onset to primary and emergency department presentations, first and diagnostic electrocardiograms, thrombolytic therapy and aspirin; drugs prescribed during hospitalisation. RESULTS: Thrombolytic therapy was given to 12/41 Indigenous patients (29%) and 38/81 non-Indigenous patients (47%) (P = 0.06). Presentation delay over 12 hours was the reason for not giving thrombolytic therapy for 14/29 Indigenous patients (48%) and 8/43 non-Indigenous patients (19%) (P < 0.01). Median delay times were longer for Indigenous patients for all six categories of delay, although the difference was significant only for delay to emergency department presentation (10:00 versus 3:26 hours; P < 0.01) and to diagnostic electrocardiogram (8:10 versus 3:50 hours; P < 0.01). Delays were also longer for patients from rural compared with urban areas. Once diagnosed, Indigenous patients were as likely as non-Indigenous patients to receive aspirin (93% versus 96%) and beta-blockers (70% versus 69%) and more likely to receive angiotensin converting enzyme inhibitors (60% versus 40%; P = 0.03). CONCLUSIONS: Delays in presentation affect Indigenous people living in rural and urban areas as well as non-Indigenous people living in rural areas. Concerted efforts are needed to improve health service access in rural areas and to encourage Indigenous people with persistent chest pain to present earlier. PMID- 11008595 TI - Imagined ugliness: a symptom which can become a disorder. AB - Concern regarding physical appearance is normal, but can sometimes reach an intensity which renders it a "disorder" (body dysmorphic disorder, or BDD). The rate of BDD in the general population is not clear, but sufferers are over represented in psychiatric populations and in those seeking cosmetic surgery. BDD shows extensive psychiatric comorbidity, including depression, social phobia, and obsessive-compulsive disorder. BDD is treatable, with serotonergic antidepressants (e.g., selective serotonin reuptake inhibitors) and cognitive behavioural treatments being the most effective. PMID- 11008596 TI - The clinical role of quantitative ultrasound in assessing fracture risk and bone status. AB - Quantitative ultrasound (QUS) analysis of calcaneal bone is useful to quantify fracture risk, and in monitoring response to treatment and identifying bone changes associated with disease and medication. However, diagnosis of osteoporosis by QUS measurements remains contentious, but the problems are due to the limitations of the present T-scores rather than the technique. QUS offers the potential for the widespread detection of low bone mass and subsequent management and prevention of fracture. PMID- 11008597 TI - The new health insurance rebate: an inefficient way of funding public hospitals. PMID- 11008598 TI - Use of diagnosis codes to understand variations in hysterectomy rates: a pilot study. PMID- 11008599 TI - Aeromonas hydrophila infection secondary to an electrical burn. PMID- 11008600 TI - The hospitalist: a third alternative. PMID- 11008601 TI - Use of antibiotics in the management of prelabour rupture of the membranes at term. PMID- 11008602 TI - Universal periconceptional folate supplementation: chasing a dream? PMID- 11008603 TI - The Declaration of Helsinki and research in vulnerable populations. PMID- 11008604 TI - Caring in healthcare. PMID- 11008605 TI - Is there a place for a graduate entry medical course in New Zealand? PMID- 11008606 TI - Inequalities: a challenge to science and politics. PMID- 11008607 TI - Plastic wrapping of cot mattresses: results from a pilot study. AB - AIM: To assess the prevalence of plastic wrapping of cot mattresses and their thickness. METHODS: Mothers of infants less than six months attending Plunket clinics in Central Auckland were interviewed. The thickness of the plastic was measured. RESULTS: 99 of 110 (90%) mothers invited to participate were visited at home. The sample was socioeconomically advantaged. Most infants were breastfed and few slept prone. The prevalence of plastic wrapped cot mattresses was 23.2%, of which sixteen (out of 23) used BabeSafe and seven used other types. The mean thickness of the BabeSafe was 0.15 mm (range 0.12-0.19 mm) and the other types were 0.10 mm (range 0.04-0.13 mm). One sample of plastic was 0.04 mm. CONCLUSION: Thin plastic wrapping is being used and is potentially dangerous. PMID- 11008608 TI - A decade of intensive care unit trauma admissions in Auckland. AB - AIMS: To describe the demographics, nature and severity of injury of trauma admissions to a New Zealand urban Intensive Care Unit (ICU) over a ten year period; to determine differences in injury characteristics between patients received from inside and outside the local trauma catchment area; and to calculate incidence rates in the local population served, to identify high risk groups of patients. METHODS: We carried out a cross-sectional analysis of a prospective ICU patient registry. Data on all trauma admissions from 1988 to 1997 to the ICU of a large New Zealand urban hospital were studied with respect to age, gender, ethnicity, injury type and severity, and referral status. National Census data for the ICU catchment area were used to calculate incidence rates for local admissions. RESULTS: A total of 2305 trauma patients were admitted over the period of the study, accounting for 25% of all ICU admissions. The median age was 28 years and 75% were males. Blunt trauma, mostly due to motor vehicle crashes, accounted for 95% of admissions and penetrating trauma was very rare. The median Injury Severity Score (ISS) was 26 and most life threatening injuries occurred in the head region. Referred admissions were more severely injured and had a higher prevalence of severe head injury than local admissions. The ICU trauma admission rate for local patients was 34.6 per 100,000 person-years. Males had a higher rate than females in all age groups. New Zealand Europeans made up the majority of admissions, but Maori and Pacific Island males had the highest incidence rates. CONCLUSIONS: This study identified trauma as a major component of the ICU population. ICU trauma admissions were characterised by young males with head injuries resulting from motor vehicle crashes. Referred admissions were more severely injured than local admissions and would thus distort the estimated incidence of trauma in the local geographic region served. Calculation of incidence rates highlighted a significantly higher risk of ICU trauma admission amongst Maori and Pacific Islands people than New Zealand Europeans. PMID- 11008609 TI - Risk assessment in upper gastrointestinal haemorrhage: implications for resource utilisation. AB - AIMS: Acute upper gastrointestinal (GI) bleeding is a common and serious problem with an associated mortality of some 10%. It is desirable, both for optimising outcomes and for the efficient use of resources, that some form of risk assessment be made early and management be stratified accordingly. A risk scoring system was recently proposed and validated by Rockall and colleagues from the UK. We wished to assess its validity in a New Zealand setting. METHODS: 565 consecutive patients treated for acute upper gastrointestinal bleeding at Wellington Hospital between 1988 and 1991 were the subject of a major prospective study. All patients were retrospectively assigned a score of 0-7 based on presentation criteria as suggested by Rockall and colleagues. The score is a composite one having regard to age, haemodynamic variables on presentation and associated serious co-morbidity. Correlation was sought between the score and the in hospital mortality risk. RESULTS: The overall 30-day mortality was 11%. Of the 60.5% of patients with a total score of less than 4 ('low risk'), the group mortality was 3.2%. Of the 39.5% of patients with a total score of 4-7 ('high risk'), the group mortality was 22.4%. CONCLUSIONS: The scoring system appears as valid in a New Zealand patient population as in the UK. We suggest that this scoring system be adopted in hospitals for patients with acute upper GI bleeding to efficiently direct the use of 'intensive care' type facilities to those most at risk, and thereby optimise management. PMID- 11008610 TI - Posterior urethral valves: failure of antenatal diagnosis. PMID- 11008611 TI - Work capacity evaluation. PMID- 11008612 TI - Inhaled steroids for chronic obstructive pulmonary disease--where are we now? PMID- 11008613 TI - Market testing the testers: lessons from recent New Zealand clinical laboratory management strategy. PMID- 11008614 TI - Report of the Cancer Control Workshop '99. Members of the National Cancer Control Steering Committee. PMID- 11008615 TI - Queensland tick typhus infection acquired whilst on holiday in Queensland. PMID- 11008616 TI - The effectiveness of postal dissemination of evidence-based clinical guidelines. PMID- 11008617 TI - British Toxicology Society Annual Congress. University of York, 26-28 March 2000. Abstracts. PMID- 11008618 TI - 31st Malaysia-Singapore Congress of Medicine. 20-24 August 1997. Abstracts. PMID- 11008619 TI - 3rd Meeting of the Asian Society of Cardiothoracic Anesthesia and 11th ASEAN Congress of Anesthesiologists. 7-11 July 1999. Abstracts. PMID- 11008620 TI - International Conference in Gastroenterology. 23-25 July 1999. Abstracts. PMID- 11008621 TI - Annual meeting of the Canadian Society for Clinical Investigation, The Royal College of Physicians and Surgeons of Canada, and participating societies. Edmonton, Alberta, Canada. September 21-24, 2000. Abstracts. PMID- 11008622 TI - 36th Annual meeting of the European Association for the Study of Diabetes. Jerusalem, Israel, 17-21 September 2000. Abstracts. PMID- 11008623 TI - 15th Annual meeting of the European Society of Human Reproduction and Embryology and annual meeting of the French Federation for the Study of Reproduction. Tours, France, 26-30 June 1999. Abstracts. PMID- 11008624 TI - 22nd Annual meeting of the American Society for Bone and Mineral Research. Toronto, Ontario, Canada. September 22-26, 2000. Abstracts. PMID- 11008625 TI - VIII International Conference on Inborn Errors of Metabolism. Cambridge, United Kingdom, 13-17 September 2000. Abstracts. PMID- 11008626 TI - Psychoanalysis at the millennium. AB - This article explains and discusses the immense complexity of the psychoanalytic process as it is becoming increasingly understood at the millennium, and offers the possibility that it can be viewed from at least five channels of psychoanalytic listening. The careful ongoing examination of the transference countertransference interactions or enactments, and their "analytic third" (32) location in the transitional space is extremely important in psychoanalytic practice. We must be careful in our interpretations of the clinical data not to stray any farther from the fundamental concepts of Freud than is necessary, lest we end up with a set of conflicting speculative metaphysical systems and become a marginalized esoteric cult. Freud's work remains our basic paradigm, the core of psychoanalysis, even though his papers on technique and his emphasis on the curative power of interpretation are from a one-person psychology standpoint and his view of psychoanalysis as just another empirical 19th-century science requires proper understanding and emendation in the light of accumulated clinical experience since his time. PMID- 11008627 TI - Cognitive therapy of schizophrenia: a new therapy for the new millennium. AB - Over the past decade, major advances have been made in extending the principles and therapeutic strategies of cognitive therapy to the treatment of schizophrenia. In a number of large-scale outcome studies with cognitive therapy for schizophrenia, cognitive therapy has been shown to offer significant gains for those patients who have not been wholly helped with medications. It may even serve to prevent the consolidation of the illness if delivered with those in the early stage of the illness. We first outline the "state-of-the-art" conceptualization and strategies employed by cognitive therapists to treat positive and negative symptoms and then review the clinical trials. PMID- 11008628 TI - Preserving the "psychosocial" in an era of biological psychiatry. AB - For psychodynamic thinking and understanding to feel relevant to psychiatric residents in current training programs, it must focus on the clinical problems they encounter with patients. It must be presented in everyday language and in a form applicable to their own clinical experience. Abstract theory or "deep" formulation may alienate rather than attract interest. PMID- 11008629 TI - The application of computerized content analysis of natural language in psychotherapy research now and in the future. AB - For many years the author and his colleagues have been involved in studying the roots and processes of the conveyance of semantic messages via spoken language and verbal texts. After establishing that reliable and valid measurements of highly relevant neuropsychiatric categories, such as anxiety, depression, and cognitive impairment, can be made by identifying and counting the occurrence per grammatical clause of language content and form categories typifying specific content-analysis scales, the research focus has turned towards computerizing this process of content analysis. This report summarizes the achievements and applications of the current empirical status of this method of computerized content analysis of natural language to psychotherapy research, and it speculates on possible future applications in the millennium. PMID- 11008630 TI - New trends for a new millennium. AB - Six new trends leading us into the new millennium are described. They include psychotherapy integration, specialization, managing care, the expanding scope of practice, cultural diversity, and psychotherapy without walls. We are entering an age of greater public access to psychotherapeutic interventions, and more control of our professional field. Changes will be revolutionary. PMID- 11008631 TI - Object relations and intersubjectivity in depression. AB - Starting with a clinical vignette the authors discuss Freud's thesis from "Mourning and Melancholia" (1917) that depression is a defense against the experience of loss: Whilst a part of the self remains identified with the lost object another part directs all the aggression against it that originally had been directed against the object. As a result the relationship between self and object becomes replaced by a pathological relationship between parts of the self. As psychoanalytical and phenomenological-hermeneutical approaches congruently show, one reason why mourning cannot proceed and feelings of guilt cannot be worked through lies in the fact that depressive patients need a symbiotic, idealizing type of identification with their objects in order to stabilize their identity. Because of their narcissistic vulnerability changes, personal failures, and disappointment by their objects, which may provoke unconscious aggression and fear of loss, can easily threaten their psychic equilibrium. This gives rise to considerable difficulties within the transference-countertransference relationship. The authors argue that to establish a feeling of autonomy in depressive patients, a secure framework within therapy has to be offered and at the same time the negative transference must thoroughly be worked through. PMID- 11008632 TI - Neurological dysfunction, psychic conflict, and psychotherapy. AB - Clinical experience of psychotherapists is getting more and more circumscribed by the boundaries of theoretical references. The Freudian "psychic treatment" for neurosis got its impetus from the friendship between Sigmund Freud and Ludwig Binswanger, the phenomenological psychiatrist. This relationship laid the foundation for the references of psychoanalytic psychotherapy in Europe. Then, the Freudian conception of actual neurosis led to a psychosomatic approach. Later, the study of narcissistic neurosis opened up to borderline patients. Today, scientific and medical progress greatly influence psychopathological research and the way we look at our patients and their requests. These changes come from the patients' sayings and their expectancy of well-being. At the same time psychotherapists have to take a new look at their theoretical references. On the one hand, a dynamic concept for psychotherapy is necessary for integrating the phenomenologic approach into psychoanalysis, and providing the understanding of situations emerging in neurological dysfunction. On the other hand, psychotherapists pay more attention to the different actors contributing to a pathology (Who is suffering? What about the life-partner or the relatives?). These changes lead to a new look into the processes of identification as well as the notion of identity. This article discusses these influences on psychotherapy and clinical research showing how clinical situations get ahead of theoretical references. PMID- 11008633 TI - Presentness: an intersubjective dimension of the therapeutic act. AB - In this article, the author discusses a mode of the therapist's presence as an important dimension within the intersubjective framework. She seeks to introduce the term presentness to denote dimensions of the therapist's explicit usage of her implicit knowledge and the role of her unformulated experiences within the therapeutic situation. The emphasis is on the power of these shared states of mutual reverie and moments of spontaneous responsiveness on the part of the therapist. The author wants to emphasize again that this term describes an important, though not exclusive, dimension of the therapeutic interaction. It does not replace such psychic functions as observation, distancing, empathy, or such modes of object-relating as transference, countertransference or projective identification. The term presentness illuminates the symmetrical dimension of the therapeutic interaction, and its contribution to the construction of "meaningfulness" for both patient and therapist. PMID- 11008634 TI - The birth of reality: psychoanalytic developmental considerations. AB - In this article, utilizing an emended version of Winnicott's notion of transitional objects and recent parent-infant research, the author offers a perspective on the development of individuals' experience of being alive and real in relation to objects and persons. Primary transitional objects, which represent parent-infant interactions, facilitate the infant's transition from early undifferentiated, bodily, and global experiences of being real and alive to the infant's yoking and extending these subjective organizations to recognized not-me objects. Secondary transitional objects make their appearance when the child begins to acquire the capacities for self-reflexivity, symbolization, and language. These objects provide the child with opportunities to gain confidence and courage in extending, yoking, and making use of his/her subjective experiences of being alive and real in relation to cultural symbols and rituals. More importantly, secondary transitional objects are paths toward mutual recognition, acceptance, and sharing experiences of being alive and real. PMID- 11008635 TI - Second generation to Holocaust survivors: enhanced differentiation of trauma transmission. AB - "Second generation to Holocaust Survivors" is a description of a segment of society, as well as an attempt at characterizing these individuals. It is common to speak of mechanisms of transmission of trauma as characterizing the dynamics of the second generation. This paper intends to advance differentiation between two kinds of transmission of trauma: direct transmission (also called transposition) and indirect transmission. There seems to be some confusion in this realm, since there is some discrepancy between clinical and the experimental publications: Whereas the first usually presents evidence of direct transmission of trauma in the second generation, the second mostly demonstrates indirect transmission. We shall present a clinical account of group therapy demonstrating indirect transmission, proposing a distinction between second-generation individuals owing to the relative dominance of each of the mechanisms in their mental structuring. This distinction has significant clinical consequences. PMID- 11008636 TI - Psychotherapy with older adults. AB - The scope of need and possible benefit of psychosocial interventions in late life far exceeds the scientific evidence of efficacy. Most studies have been conducted in academic settings with self-selected, relatively independent older adults for whom measures of benefit were narrowly defined. In reviewing the existing literature concerning individual, group, and couples therapies in the geriatric population, we find that with appropriate adaptations for medical comorbidity, cognitive and sensory impairment, caregiver inclusion, and realistic modification of treatment goals, psychotherapeutic endeavors with the senior patient can prove to be highly successful. PMID- 11008637 TI - Transpersonal psychotherapy. AB - The history, theory, and practice of Transpersonal (or Spiritual) Psychotherapy are presented. The author describes his own evolution from a traditional psychoanalyst to a psychotherapist who uses the tools and wisdom from spiritual traditions to enhance traditional psychotherapy while, at the same time, improving the self system of the therapist. Dangers as well as benefits of the spiritual approach are outlined. The creation and holding of a spiritual or transpersonal context is described and ways to ascertain, in the clinical situation, the appropriateness of such an approach are explained. The use of bibliotherapy to help transform and expand the worldview of the patient is outlined. Prayer and meditational systems also have a healing role in this approach. To illustrate the uses of Transpersonal Psychotherapy in practice, four cases are presented: 1) a paranoid schizophrenic man, 2) a well-functioning borderline person, 3) a very poorly functioning borderline person, and 4) a high functioning neurotic man who had been in psychoanalysis. PMID- 11008638 TI - Narrative lessons for the psychotherapist: Kafka's the metamorphosis. PMID- 11008639 TI - [Periodate oxidation in chemistry of nucleic acid. Dialdehyde derivatives of nucleosides, nucleotides, and oligonucleotides]. AB - The employment of periodate oxidation in the chemistry of nucleic acids and their components is reviewed. The reaction mechanism, structural requirements to substrates, and synthesis of dialdehyde derivatives of nucleosides, nucleotides, and oligonucleotides are discussed in the first part. The second part involves chemical, physicochemical, and biological properties of the dialdehyde derivatives, as well as their use for the affinity modifications of proteins. PMID- 11008640 TI - [Biologically active fragment of the differentiation factor from HL-60 cell line. Identification and properties]. AB - Six-membered peptide fragment TGENHR (HLDF-6) was identified in the HL-60 cell culture of human promyelocyte leukemia treated with retinoic acid when studying the differentiation factor HLDF of this cell line. HLDF-6 retains the ability of the full-size factor to induce the differentiation and arrest the proliferation of the starting HL-60 cells. It was shown that the synthetic peptide HLDF-6 has no specific receptors on the surface of the HL-60 cells but can affect the binding of interleukin IL-1 beta, a cytokine involved in proliferation, to the cell surface. It was found on a model of transplantable NSO myeloma that HLDF-6 has an antitumor activity. PMID- 11008641 TI - [Solid phase catalytic hydrogen isotope exchange in dalargin]. AB - A [3H]Dalargin preparation with a molar radioactivity of 52 Ci/mmol was obtained by the high temperature solid-state catalytic isotope exchange (HSCIE) of tritium for hydrogen at 150 degrees C. This tritium-labeled peptide was shown to completely retain its biological activity in the test of binding to opioid receptors from rat brain. The dissociation constant of the Dalargin-opioid receptor complex was found to be 4.3 nM. The dependencies of the chemical yield and the molar radioactivity on the reaction time and temperature of HSCIE were determined. The activation energy of the HSCIE reaction for the peptide was calculated to be 32 kcal/mol. The amino acid analysis showed that tritium is distributed between all the amino acid residues of [3H]Dalargin at the HSCIE reaction, with the temperature growth significantly increasing the total tritium incorporation and, especially, enhancing the radioactivity incorporation into aromatic residues. PMID- 11008642 TI - [Analytical biotechnology of recombinant peptides and proteins. II. Primary structure of the fusion protein containing human proinsulin and optimization of its proteolysis by trypsin]. AB - The kinetics of trypsin proteolysis of the fusion protein (FP) containing human proinsulin was studied by a set of analytical micromethods. These were the microcolumn reversed-phase HPLC and the qualitative identification by MALDI-TOF mass spectrometry and amino acid sequencing. The first stage of the proteolysis was shown to be the cleavage of FP into the leader fragment and proinsulin. The subsequent splitting off of C-peptide from proinsulin results in the formation of ArgB31-ArgB32-insulin. The effect of temperature on the formation of de-ThrB30 insulin, a by-product, was also studied. The structure of FP was confirmed by the peptide mapping technique, and the leader fragment was shown to contain no N terminal Met residue. PMID- 11008643 TI - [Hydrolysis of chimeric proteins by enteropeptidase at the specific linker (Asp)4Lys depending on refolding conditions]. AB - Refolding from inclusion bodies of chimeric proteins containing the enteropeptidase-specific linker (Asp)4Lys was carried out. It was shown that, depending on the refolding conditions, chimeric proteins function as substrates or inhibitors of the enteropeptidase. The efficiency of the enteropeptidase hydrolysis of chimeric proteins containing the (Asp)4Lys linker may depend not only on the amino acid sequence of the protein binding site for the enzyme but also on the site conformation. PMID- 11008644 TI - [Coupling of proteolysis with ATP hydrolysis by Escherichia coli Lon proteinase. I. Kinetic aspects of ATP hydrolysis]. AB - Some aspects of the ATPase function of the Escherichia coli Lon protease were studied around the optimum pH value. It was revealed that, in the absence of the protein substrate, the maximum ATPase activity of the enzyme is observed at an equimolar ratio of ATP and Mg2+ ions in the area of their millimolar concentrations. Free components of the substrate complex (ATP-Mg)2- inhibit the enzyme ATPase activity. It is hypothesized that the effector activity of free Mg2+ ions is caused by the formation of the "ADP-Mg-form" of the ATPase centers. It was shown that the activation of ATP hydrolysis in the presence of the protein substrate is accompanied by an increase in the affinity of the (ATP-Mg)2- complex to the enzyme, by the elimination of the inhibiting action of free Mg2+ ions without altering the efficiency of catalysis of ATP hydrolysis (based on the kcat value), and by a change in the type of inhibition of ATP hydrolysis by the (ADP Mg)- complex (without changing the Ki value). Interaction of the Lon protease protein substrate with the enzyme area located outside the peptide hydrolase center was demonstrated by a direct experiment. PMID- 11008645 TI - [Isotyping of human C4 complement using differences in the functional activity of C4A and C4B isotypes]. AB - The difference in the functional activity of the isotypes A and B of component C4 of human complement was used to determine their ratio and to detect the inherited deficiency of the isotypes. ELISA methods were developed for the quantitative assay of component C4 (conventional sandwich method) and its functional activity. When determining the functional activity, the classic pathway of the complement and therefore of component C4 was activated by activators sorbed on ELISA microplates (immunoglobulin IgG3 or liposaccharide of the Shigella sonnei cell walls, which activates the complement by binding component C1). The nascent fragment C4b is covalently bound to the target activator; C4Ab binds better to the target protein (immunoglobulin), and C4Bb to the target carbohydrate (liposaccharide). Therefore, when immunoglobulin is a target activator, isotype C4A is bound and determined; and when the complement is activated by liposaccharide, isotype C4B is determined. The ratio of the activities determined by the two methods indicates a deficiency in the individual isotypes of component C4 or its absence. The rabbit polyclonal monospecific antibodies against the human component C4 and the conjugates of these antibodies with horseradish peroxidase were used in the methods described. PMID- 11008646 TI - [Turbidimetric study of latex agglutination in 2,4-dichlorophenoxyacetic acid analysis]. AB - By the example of 2,4-dichlorophenoxyacetic acid (2,4-D) assay by the inhibition of latex agglutination, new synthetic polymer microspheres for the conjugation with antibodies to 2,4-D and agglutinators based on ovalbumin and polyacrylamide were developed and characterized. The effect of various parameters (the concentration of reagents, the type and the degree of modification of the microsphere surface, and the nature of the carrier in the composition of the agglutinator) on the rate of agglutination and the maximal optical absorption observed during the reaction were studied by turbidimetry. The optimal parameters were found for the assay of 2,4-D by the inhibition of latex agglutination with turbidimetric registration of the results. PMID- 11008648 TI - Social networks and the elderly: conceptual and clinical issues, and a family consultation. AB - After a general introduction to the construct "social networks," this article discusses the progressive transformation of the personal social network--family, friends and acquaintances, work and leisure relationships, et cetera--as individuals reach an advanced age. This is followed by a summary and discussion of a clinical consultation, with an emphasis on the reciprocal influence between individual and social network. PMID- 11008649 TI - Tales full of sound and fury: a cultural approach to family therapeutic work and research in rural Scandinavia. AB - The concept of "culture" figured prominently in the development of family therapy. Recent conceptualizations, however, have tended to focus primarily on the ideational dimensions of culture. While not disputing that meanings and other ideas constitute significant features of group lifeways, this article proposes a return to earlier anthropological framings that incorporate material and ideational dimensions of cultures. To illustrate how his expanded concept may serve as a guide for therapeutic work, the article describes therapy with one family at a clinic in rural Scandinavia. We especially focus on the place of key symbols as historical links between the ideational and material dimensions of cultures. The perspective developed here is one of seeing cultures as sets of interpenetrating actions and ideas shaped by as well as shaping their practitioners. PMID- 11008650 TI - The deconstruction of magic: rereading, rethinking Erickson. AB - This article examines the case history writings of Milton Erickson as literary texts. A deconstructive analysis reveals that the modernist assumption of therapist as magical healer disguises the role clients play in their own change. Hypothetically constructed case transcripts contrasted with excerpts from Erickson (see Rossi, 1980) illustrate how the inclusion of client voice enriches understanding of therapeutic change. PMID- 11008651 TI - Evolving the biobehavioral family model: the fit of attachment. AB - The 1993 Biobehavioral Family model (BBFM) posits that family relational patterns and biobehavioral reactivity interact so as to influence the physical and psychological health of the children. The revised 1999 BBFM incorporates parent child attachment as a pivotal construct. The current study tests the 1999 BBFM by predicting, in asthmatic children, that child perception of parental relationship quality, triangulation of child in marital conflict, and parent-child security of relatedness will be associated with hopelessness and vagal activation (one mechanism of airway compromise in asthma). In this study, 22 children with asthma (11 males/11 females, aged 8 to 16), watched, alone, an emotionally challenging movie, then engaged in family discussion tasks (problem solving, loss, conflict, cohesion) and completed the Children's Perception of Interparental Scale, the Relatedness Questionnaire, The Multidimensional Scale of Anxiety in Children, and the Hopelessness Scale for Children. Heart rate variability, measured at baseline and throughout the movie and family tasks, was used to compute respiratory sinus arrhythmia (RSA)--an inferential measure of vagal activation. The child's perception of parental conflict showed trends of association with triangulation and insecure father-child relatedness. Triangulation and hopelessness also were associated with insecure father-child relatedness, all of which were associated with vagal activation. Insecure mother-child relatedness was correlated only with hopelessness. Anxiety was not related to any variables. These findings lend support to the 1999 BBFM, and suggest a key role for parent-child attachment. PMID- 11008652 TI - Empathy in families of women with borderline personality disorder, anorexia nervosa, and a control group. AB - This is a study of empathy in the families of 27 women with borderline personality disorder (BPD), 28 women with restricting anorexia nervosa (AN), and 27 women without a clinical diagnosis (NC). The daughters and both parents responded to the Interpersonal Reactivity Index (IRI), an instrument for assessing four dimensions of empathy. In addition, they were personally interviewed, with the Family Interview for Protectiveness and Empathy (FIPE), about the extent of empathy expressed by the parents to their daughter during her development. On the IRI, women with BPD scored highest on the immature and lowest on the mature aspects of empathy, whereas scores of AN and NC women were all within normal limits. Parents of BPDs had the lowest IRI scores, while parents of AN and NC groups were similar to each other and to criterion group scores. IRI scores of AN daughters were positively correlated with their parents' scores whereas BPDs' scores were negatively correlated with those of their parents. There were no correlations between the IRI scores of NC subjects and their parents. On the FIPE, borderline daughters and parents agreed about the relative absence of empathic parenting, whereas AN and NC daughters and parents agreed as to the presence of empathic parenting. The theoretical and clinical implications of these contrasting findings are discussed. PMID- 11008653 TI - Family perception of interpersonal behavior as a predictor in eating disorders: a prospective, six-year followup study. AB - This study assessed family perception patterns of interpersonal behavior in families with a daughter diagnosed with an eating disorder 6 years after treatment that used a prospective design. Family perception patterns of patients found to have a poor outcome at followup (n = 15) were compared with patients with a good outcome (n = 23), as well as a control group (n = 36). Using the system of multiple level observation of groups (SYMLOG), all 238 family members evaluated themselves and each other. The index patients with a poor outcome perceived themselves as friendlier and more positive than they were perceived by their families. In comparison to parents of daughters with a good outcome and the control group, parents of daughters in the poor outcome group perceived themselves to be less friendly than their partners perceived them to be. A rigid polarization of the perception of the index patient by family members and a discrepancy in this perception between the index patient and the rest of the family were found to be indicative of a poor prognosis. Implications for treatment based on family perception patterns of interpersonal behavior are discussed. PMID- 11008654 TI - Further evidence that the family assessment device should be reorganized: response to Miller and colleagues. AB - Ridenour, Daley, and Reich's (1999) factor analyses of the Family Assessment Device (FAD) indicated that the FAD be reorganized. Miller and colleagues (2000) contend that the FAD be used in its original format based on the FAD's theoretical foundation; however, this does not preclude the importance of scientific scrutiny of an instrument's construct validity to determine how well an instrument represents its theoretical base. Subscale factor analyses (exploratory and confirmatory), item-level factor analysis, and the clinical and psychometric studies cited by Miller and colleagues suggest a more parsimonious FAD configuration and were consistent with Ridenour and colleagues' factor analyses. PMID- 11008655 TI - Why fix what isn't broken? A rejoinder to Ridenour, Daley, & Reich. AB - Ridenour, Daley, & Reich (2000) suggest that the Family Assessment Device should be reorganized. We disagree and provide further reasons why such a reorganization is unwise. PMID- 11008656 TI - M. leprae-HIV co-infection: pattern of immune response in vivo and in vitro. PMID- 11008657 TI - The source of infection: an unsolved issue. PMID- 11008658 TI - Cytokine patterns using molecular tools. PMID- 11008659 TI - Prospects for chemotherapy of leprosy. PMID- 11008660 TI - Clinical leprosy through the last seventy-five years. PMID- 11008661 TI - An epidemiological analysis of leprosy from 1951-1996 in Sichuan. AB - Through the leprosy surveillance system established in 1990s the authors analysed the epidemiology of leprosy in Sichuan province from 1951 to 1996. By the end of 1996, the total number of registered cases was 32,772, the peak incidence rate (2.44/100,000) was in 1958. The annual average progressively decreasing speed (AAPDS) of the number of new cases, incidence rate, incidence rate among children, number of newly detected patients, detection rate, number of registered cases and registered prevalence rates were 4.7% (xg = 0.9534), 5.9% (xg = 0.9407), 7.4% (xg = 0.9263), 6.7% (xg = 0.9326), 8.1% (xg = 0.9195), 10.9% (xg = 0.8913) and 11.7% (xg = 0.8828) respectively. Among newly detected cases, the proportion of children gradually declined with an AAPDS of 2.9% (xg = 0.9712), but that of household contacts gradually increased, showing a gradual decline of source of infection. The multibacillary (MB) rate among new cases, newly registered cases and active cases also increased gradually. The MB rate among child cases was much lower than that among adults. The disability rates among newly registered cases tended to decline. The peak incidence was in the 20 to 35 years age group, the average age at onset being 31.92 years. Average age at onset gradually increased to 34.19 during 1991-1995. The average interval between onset of symptoms and diagnosis was 4.89 years and that had reduced gradually to 3.24 years during 1991-1995. The authors conclude that: (1) the epidemiological trend of leprosy in Sichuan province shows decline; (2) the case-finding activities have improved and intensified; but (3) the disability rate among newly detected cases is still high (> 20%) and the average delay in diagnosis is still too long, showing that early case finding is still not satisfactory. PMID- 11008662 TI - Techniques for covering soft tissue defects resulting from plantar ulcers in leprosy: Part V. Use of the flap in the inguinal region and latissimus dorsi musculocutaneous flap. AB - The area of distribution of the superficial circumflex iliac, superficial epigastric and superficial external pudental arteries is large and flaps based on them can meet the requirement of different recipient sites. We have transplanted free flaps based on the superficial epigastric artery for repairing plantar soft tissue defects in six leprosy patients. During the follow-up examination 58 to 118 months later there has been no recurrence of ulceration in any of these cases. The latissimus dorsi muscle, is mainly nourished by the thoracadorsal artery and the latissimus dorsi musculocutaneous flap is a large sized, composite structure with abundant blood provision and strong anti-infectious property. The latissimus dorsi flap can be used as an artery-pedicled island flap or as a free flap besides its use as a muscle graft, because of its constant vascular position, wide outer-diameter of the vessels and long pedicle. It can therefore be utilized for repairing soft tissue defect or replacement of paralyzed muscle. We have used the latissimus dorsi musculocutaneous free flap for repairing large skin and soft tissue defects resulting from plantar ulceration in three leprosy patients. During the follow-up period, one patient who had complete drop-foot and had refused corrective surgery had recurrence of the ulcer in the 12th post operative month. No ulcers had recurred in the other two cases during the follow up at 48 and 114 months. PMID- 11008663 TI - Leprosy presenting as macrocheilia. PMID- 11008664 TI - Modification of the "wrap around" tendon anastomosis of fascia lata graft with a slim double tendon of palmaris longus. PMID- 11008665 TI - Chemotherapy in paucibacillary leprosy. PMID- 11008666 TI - Leprosy patients and their spouses. PMID- 11008668 TI - Alternative strategy to total population survey. PMID- 11008667 TI - Community based leprosy survey in U.T. Chandigarh. PMID- 11008669 TI - Elimination of leprosy--what should it mean and what should we do: a personal view. PMID- 11008670 TI - Where's the line? PMID- 11008672 TI - Improving mental health professional-client relationships. PMID- 11008671 TI - Storytelling: using an ancient art to work with groups. PMID- 11008673 TI - Resiliency is key for children to overcome adversity. PMID- 11008674 TI - Fall incidence in a state psychiatric hospital in Singapore. AB - The results of a retrospective study of fall incidence during a 1-year period in a psychiatric hospital in Singapore are reported, involving 309 patients who fell one or more times during their stay. The profile of inpatients involved in falls was identified. Data were derived from standard incident forms completed whenever patients had a fall. A total of 348 falls were identified for the 1-year period. Results revealed higher fall rates in younger male epileptic patients; elderly female patients, age 70 and older with depression or dementia; individuals with concomitant medical conditions; and patients taking three types of medication. The majority of falls occurred when the activity level was high in the ward. Young epileptic patients and elderly depressed patients were prone to repeated falls. These results have the potential to assist in identifying patients at high risk and in designing and implementing strategies to prevent such incidents. PMID- 11008675 TI - Facilitating a multidisciplinary parent support & education group guided by Allen's developmental health nursing model. AB - Parents are generally the primary source of support and nurturing for children experiencing an eating disorder; yet, they often feel blamed for the illness. Guided by the developmental health model, the group focused on parents' competence and helped them share solutions; reframed situations related to development in adolescence and mid-life; and shifted the emphasis to process to help parents understand behaviors. Parents became partners in helping their children recover by sharing solutions and creating learning opportunities from adversity. PMID- 11008676 TI - The history, economics, and financing of mental health care. Part 1: 17th to 19th centuries. AB - Families in colonial times cared for their mentally ill members at home, with little assistance from their communities. Community treatment is an old idea, not a new one. Early laws about containing the disturbances created by individuals with mental illness made no mention of clinical dimensions. The focus was strictly on the social and economic consequences of the mental disorders. Legislation about public mental hospitals in the mid-19th century was hardly enlightened. There were no particular plans, other than not to expend more dollars than actually necessary. PMID- 11008678 TI - What psychiatric nurses should teach the public above mental health? PMID- 11008677 TI - Combined group process-psychoeducation model for psychiatric clients and their families. PMID- 11008679 TI - [Structure and function of microbial communities in soil from the southern Taiga]. AB - General regularities in the structure of the microbial communities of southern taiga soil ecosystems and taxonomic differences between the microbial communities of soils with different hydrothermal characteristics are discussed with reference to the main types of soils of the Central State Forest Biosphere Reserve. PMID- 11008680 TI - [Synthesis of osmoprotectors by halophilic and alkalophilic methanotrophs]. AB - The 1H-NMR analysis of methanol extracts of halophilic and halotolerant alkaliphilic methanotrophs isolated from the soda lakes of Southern Transbaikal and Tuva showed that bacterial cells grown at an optimum salinity accumulated mainly sucrose and 5-oxo-1-proline, whereas cells adapted to 0.5-1.0 M NaCl additionally synthesized ectoine. A more detailed study showed that nitrogen deficiency in the growth medium of Methylobacter alcaliphilus 20Z decreased the synthesis of nitrogen-containing osmoprotectants, ectoine and 5-oxo-1-proline. M. alcaliphilus 20Z cells exhibited activities of UDP-glucose pyrophosphorylase and sucrose-phosphate synthase involved in sucrose synthesis. Glutamine synthetase in vitro did not require NH4+ ions, which implies that this enzyme is involved in 5 oxo-1-proline synthesis. Cells grown at high salinity exhibited elevated levels of aspartate kinase, aspartate-semialdehyde dehydrogenase, and ectoine synthase. This suggests that ectoine is synthesized via aspartate and aspartate semialdehyde, i.e., via the route earlier established for extremely halophilic bacteria. PMID- 11008681 TI - [Manganese-dependent ribonucleotide reductase of Propionibacterium freudenreichii subsp. shermanii: partial purification, characterization, and role in DNA biosynthesis]. AB - Like Lactobacillus leichmanii, Rhizobium meliloti, and Euglena gracilis, P. freudenreichii implicates cobalamin in DNA anabolism via adenosylcobalamin dependent ribonucleotide reductase. However, in the absence of corrinoids, P. freudenreichii is able to synthesize DNA with the involvement of an alternative ribonucleotide reductase, which is independent of adenosylcobalamin. This enzyme is localized in both the cytoplasm (80% of activity) and the cytoplasmic membrane (20% of activity), being loosely bound to the latter. Experiments with crude ribonucleotide reductase isolated from extracts of corrinoid-deficient cells showed that manganese specifically stimulates this enzyme and that it is composed of two protein subunits, a feature that is typical of all metal-containing reductases activated by molecular oxygen. Low concentrations of manganese ions enhanced DNA synthesis in corrinoid-deficient manganese-limited cells. This effect was prevented by the addition of 80 mM hydroxyurea, a specific inhibitor of metal-containing aerobic ribonucleotide reductases. It was concluded that, in adenosylcobalamin-deficient P. freudenreichii cells, DNA synthesis is provided with deoxyribosyl precursors through the functioning of manganese-dependent aerobic ribonucleotide reductase composed of two subunits. PMID- 11008682 TI - [Dependence of the effect of RNAase from Bacillus intermedius on the growth of baker's yeast on the concentration of the exogenous enzyme]. AB - Bacillus intermedius RNase added at a low concentration (0.001 microgram/ml) stimulated yeast growth, while a high RNase concentration (1500 micrograms/ml) was inhibitory to yeast growth. The inhibitory effect of RNase was transient and correlated with the increase in the trehalose pool of yeast cells. The number of unbudded cells in the yeast population tended to decrease under the action of low concentrations of bacillar RNase and to increase under the action of high concentrations of this enzyme. PMID- 11008683 TI - [Destruction of chlorinated derivatives of phenol: ortho-chlorophenol, para chlorophenol, and 2,4-dichlorophenoxyacetic acid by bacteria communities in anaerobic sludge]. AB - The bacterial community of anaerobic sludge could degrade ortho-chlorophenol, para-chlorophenol, and 2,4-dichlorophenoxyacetic acid at concentrations as high as 100 mg/l. The time needed for the degradation of a given chlorinated phenol derivative increased 1.5- to 2-fold upon a twofold increase in its concentration (from 50 to 100 mg/l). The duration of the adaptation period depended on the compound studied and on its concentration. The degradation of 2,4 dichlorophenoxyacetic acid proceeded via 2,4-dichlorophenol and p-chlorophenol as intermediates; the degradation of o-chlorophenol occurred with the formation of phenol. The dynamics of p-chlorophenol degradation and chloride ion accumulation were studied. PMID- 11008684 TI - [Growth of Rhodococcus opacus on mixtures of monohalogenated benzenes and phenols]. AB - The growth of Rhodococcus opacus GM-14 on mixtures of 2-chloro- and 2 bromophenol, of 4-chloro, 4-bromo-, and 4-iodophenol, and of chloro-, bromo-, and iodobenzenes was accompanied by consumption of the substrates and excretion of halogen ions to the medium. During the growth on monochlorophenols, the substrates were consumed sequentially in the following order: 4-chloro-, 3-chloro , and then 2-chlorophenol. Chlorine ions were excreted in a two-phase manner in amounts comprising 79% of the theoretical yield. The diauxic growth of R. opacus GM-14 can be explained by the existence in this bacterium of two modified metabolic pathways for the ortho-cleavage of halogenated pyrocatechols. The first pathway included 4-halogeno- or dihalogenopyrocatechols as intermediates, whereas the second pathway included 3-halogenopyrocatechols. PMID- 11008685 TI - [Preparation and characteristics of mutant strains of Penicillium funiculosum- producers of glucose oxidase]. AB - After the mutagenesis of Penicillium funiculosum with UV light and N-nitroso-N methylurea, 83 of 2237 grown colonies were surrounded with increased zones of glucose oxidase diffusion. Analysis of the glucose oxidase activity of selected mutant strains grown in submerged cultures allowed 18 mutant strains to be obtained whose glucose oxidase activity was 5-153% higher (in a medium with glucose) and 4-83% higher (in a medium with sucrose) than that of the parent strain. Two of these mutant strains, UV6.31 and NMU95-132, possessed high glucose oxidase activity when grown in media with glucose or sucrose and produced large amounts of mycelia. The active and morphologically stable mutant P. funiculosum NMU95-132 was chosen for further selection work. PMID- 11008686 TI - [Inorganic polyphosphates and phosphohydrolases from Halobacterium salinarium]. AB - Halobacterium salinarium grown in a liquid medium consumed up to 75% of phosphates originally present in the growth medium and accumulated up to 100 mumol Pi/g wet biomass by the time it entered the growth retardation phase. The content of acid-soluble oligophosphates in the biomass was maximum at the early stage of active growth and drastically decreased when cells reached the growth retardation phase. The total content of alkali-soluble and acid-insoluble polyphosphates changed very little throughout the cultivation period (five days). The polyphosphate content of H. salinarium cells was close to that of yeasts and eubacteria. The pyrophosphatase, polyphosphatase, and nonspecific phosphatase activities of H. salinarium cells were several times lower than those of the majority of eubacteria. The specific activity of pyrophosphatase, the most active hydrolase of H. salinarium, gradually increased during cultivation, reaching 540 mU/mg protein by the end of the cultivation period. Half of the total pyrophosphatase activity of this halobacterium was localized in the cytosol. The molecular weight of pyrophosphatase, evaluated by gel filtration, was 86 kDa. The effective Km of this enzyme with respect to pyrophosphate was 115 microM. PMID- 11008687 TI - [Features of phospholipid composition of marine proteobacteria of Pseudoalteromonas species]. AB - The study of the phospholipid composition of 14 type strains of marine proteobacteria of the genus Pseudoalteromonas showed that phospholipids are the main polar lipid constituents of membranes in these proteobacteria. The phospholipid patterns of the strains studied were found to be similar and involved five phospholipids typical of gram-negative bacteria, namely, phosphatidylethanolamine, phosphatidylglycerol, bisphosphatidic acid, lysophosphatidylethanolamine, and phosphatidic acid. The major phospholipids were phosphatidylethanolamine and phosphatidylglycerol, which add up to 89-97% of total phospholipids; bisphosphatidic acid was dominant among minor phospholipids. The prevalence of phosphatidylethanolamine (62-77% of total phospholipids) and the absence of diphosphatidylglycerol are the characteristic features of most bacteria of this genus. As in Escherichia coli, the phospholipid composition of the marine proteobacteria depended on the presence of magnesium in the medium. PMID- 11008688 TI - [Changes in the carbohydrate composition of cytosol from fungal spores in connection with environmental temperature and during storage]. AB - Differences in the carbohydrate composition were revealed among spores of fungi belonging to Zygomycetes, Ascomycota, Basidiomycota, and Oomycota, part of the novel kingdom Chromista. It was shown for the first time that Phytophthora infestans contains arabitol in addition to glucose and trehalose. Sucrose was detected in Pleurotus ostreatus basidiospores. It was established that Blakeslea trispora stylospores contain inositol. The dependence of the spore carbohydrate composition on the temperature of the habitat of the corresponding species is discussed. It was shown that the cytosol of the conidia is dominated by trehalose and inositol under hypothermic conditions and by mannitol and glucose under hyperthermic conditions. Neomycota and Eomycota were shown to differ in their responses to stress (starvation), which correlated with the differences in the carbohydrate composition of the spore cytosols. Assuming that cytosol carbohydrates perform a protective function, we explain the higher viability of conidia compared to stylo- and sporangiospores. PMID- 11008689 TI - [Effect of nitrogen sources on biosynthesis, chemical composition, and structure of exopolysaccharides from Aureobasidium pullulans (de Bary) Arnaud]. AB - The effect of the nitrogen source and the C/N ratio of the growth medium on the biosynthesis, composition, and structure of the exopolysaccharides (EPSs) of Aurebasidium pullulans (de Bary) Arnaud var. aubasidani Yurlova var. nov. and A. pullulans var. pullulans was studied. A. pullulans var. pullulans and A. pullulans var. aubasidani strains synthesized the maximum amounts of EPSs in the presence of, respectively, a reduced nitrogen source ((NH4)2SO4) and an oxidized nitrogen source (NaNO3) in the medium. The data presented confirm the validity of using the chemical composition and structure of the major cetavlon-precipitated fraction of A. pullulans EPSs for the characterization of intraspecies taxa. PMID- 11008690 TI - [Microbiological processes at the interface of aerobic and anaerobic waters in the deep-water zone of the Black Sea]. AB - Chemical and key microbiological processes (assimilation of carbon dioxide, oxidation and formation of methane, and sulfate reduction) occurring at the boundary between the aerobic-anaerobic interface in the deep-water zone of the Black Sea were investigated. Measurements were taken at depths from 90 to 300 m at intervals of 5-10 m. The integral rate of the dark assimilation of carbon dioxide varied from 120 to 207 mg C/(m2 day) with a maximum at the boundary of cyclonic currents. The organic matter (OM) formed from methane comprised less than 5% of the OM formed from carbon dioxide. A comparison between the rates of methane oxidation and methane production suggests that methane that is oxidized at depths from 100 to 300 m was formed in deeper water horizons. The maximum rate of sulfate reduction (1230 mg S/(m2 day)) was observed in the western halistatic region, and the minimum rate (490 mg S/(m2 day)), in the eastern halistatic region. The average rate of hydrogen sulfide production measured at three deep sea stations amounted to 755 mg S/(m2 day), or 276 g S/(m2 year). PMID- 11008691 TI - [Effect of microorganisms and seasonal factors on the isotope composition of organic carbon from Black Sea suspensions]. AB - The isotopic composition of particulate organic carbon (POC) from the Black Sea deep-water zone was studied during a Russian-Swiss expedition in May 1998. POC from the upper part of the hydrogen sulfide zone (the C-layer) was found to be considerably enriched with the 12C isotope, as compared to the POC of the oxycline and anaerobic zone. In the C-layer waters, the concurrent presence of dissolved oxygen and hydrogen sulfide and an increased rate of dark CO2 fixation were recorded, suggesting that the change in the POC isotopic composition occurs at the expense of newly formed isotopically light organic matter of the biomass of autotrophic bacteria involved in the sulfur cycle. In the anaerobic waters below the C-layer, the organic matter of the biomass of autotrophs is consumed by the community of heterotrophic microorganisms; this results in weighting of the POC isotopic composition. Analysis of the data obtained and data available in the literature allows an inference to be made about the considerable seasonable variability of the POC delta 13C value, which depends on the ratio of terrigenic and planktonogenic components in the particulate organic matter. PMID- 11008692 TI - [Potential activity of methane and ammonia oxidation by methanotropic communities from soda lakes of the southern Transbaikal]. AB - Radioisotopic measurements of the methane consumption by mud samples taken from nine Southern Transbaikal soda lakes (pH 9.5-10.6) showed an intense oxidation of methane in the muds of lakes Khuzhirta, Bumalai Nur, Gorbunka, and Suduntuiskii Torom, with the maximum oxidation rate in the mud of lakes Khuzhirta (33.2 nmol/(ml day)). The incorporation rate of the radioactive label from 14CH4 into 14CO2 was higher than into acid-stable metabolites. Optimum pH values for methane oxidation in water samples were 7-8, whereas mud samples exhibited two peaks of methane oxidation activity (at pH 8.15-9.4 and 5.8-7.0). The majority of samples could oxidize ammonium to nitrites; the oxidation was inhibited by methane. The PCR amplification analysis of samples revealed the presence of genes encoding soluble and particulate methane monooxygenase and methanol dehydrogenase. Three alkaliphilic methanotrophic bacteria of morphotype I were isolated from mud samples in pure cultures, one of which, B5, was able to oxidize ammonium to nitrites at pH 7-11. The data obtained suggest that methanotrophs are widely spread in the soda lakes of Southern Transbaikal, where they actively oxidize methane and ammonium. PMID- 11008693 TI - [Characteristics of bacterioplankton in the Loess-containing Lake Khanka]. AB - Some characteristics of bacterioplankton--generation time, daily (P) and specific (P/B) bacterioplankton production, and bacterial metabolic coefficient K2--in the loess-containing Lake Khanka were determined using five modifications of the bacterial-count procedure with the fluorescent dyes fluorescamin and erythrosin. Experiments showed that the organomineral complex (OMC) in this lake is broken down by chemoorganoheterotrophic bacteria. The increase in the loess content of the lake water intensified bacterial growth and the cycles of potassium, silicon, and other biogenic elements. The addition of starch to a loess suspension activated the breakdown of OMC due to the adsorption of starch on the OMC/water interface and stimulation of the metabolism of attached bacteria. PMID- 11008694 TI - [Spatial integration of partners and heteromorphism of cyanobacterium Nostoc muscoum CALU 304 in mixed culture with Rauwolfia]. AB - A comparative morphological study was conducted of Nostoc muscorum CALU 304 grown either as a pure culture on standard media or as a mixed culture with Rauwolfia callus tissue on a medium for plant tissue cultivation. The interaction of the cyanobacterial and plant partners results in their spatial integration into aggregates of specific anatomy, which arise periodically during the mixed culture growth. The morphology of the cyanobacterial cells varies depending on their localization in the combined aggregate. The degree of cyanobacterial heteromorphism increases with time of growth of the association. Evidence of the plant origin of the factors inducing heteromorphic changes in N. muscorum was obtained, as well as evidence indicating that these factors can rapidly diffuse in agarized medium. A conclusion is inferred that the heteromorphic cells correspond to bacterial forms that appear during unbalanced growth as an adaptation to altered environmental conditions. PMID- 11008695 TI - [Microbiological database UNIQEM: problems and prospects]. AB - The UNIQEM database, designed to accumulate microbiological data of a general nature, is currently used to store and make available information about microorganisms studied and maintained at the Institute of Microbiology, Russian Academy of Sciences. UNIQEM is composed of two related databases. The first one is a catalogue database, used to accumulate and maintain list-form information on a wide range of microorganisms. The second one is a property database, which facilitates collecting, processing, and publishing diverse data having to do with these microorganisms and their properties. The database supports the retrieval of microorganisms by specifying an arbitrary set of their properties and has the potential for eventually evolving into a computer instrument for unattended identification of microorganisms. PMID- 11008696 TI - [Microsampling of rhizosphere soil and laboratory artefacts]. AB - A parallel analysis of the microsamples of surrounding soil and microsamples of rhizosphere soil did not reveal the so-called rhizospheric effect. The data obtained showed that dilution significantly influences the results of determination of the number of soil microorganisms. The actual number of microorganisms revealed in soil samples greatly differed from the theoretically predicted values. The enumeration of microorganisms in soil microsamples by direct count and, especially, by the plating method with the use of conversion coefficients based on the degree of sample dilution gave erroneous results. PMID- 11008697 TI - [Glucuronate media for isolating tremelloid yeasts]. AB - D-Glucuronate-containing agar is suggested for evaluation the population density and diversity of tremelloid yeasts in natural cenoses. This medium is superior to the commonly used wort agar on which many representatives of tremelloid yeasts cannot be revealed. PMID- 11008698 TI - Endocrine disruptors: present issues, future directions. AB - A variety of natural products and synthetic chemicals, known collectively as endocrine-disrupting compounds (EDCs), mimic or interfere with the mechanisms that govern vertebrate reproductive development and function. At present, research has focused on (i) the morphological and functional consequences of EDCs; (ii) identifying and determining the relative potencies of synthetic and steroidal compounds that have endocrine-disrupting effects; (iii) the mechanism of action of EDCs at the molecular level; and (iv) the recognition that in "real life," contamination usually reflects mixtures of EDCs. Future research must examine (i) the interactive nature of EDCs, particularly whether the threshold concept as developed in traditional toxicological research applies to these chemicals; (ii) when and how EDCs act at the physiological level, particularly how they may organize the neural substrates of reproductive physiology and behavior; (iii) the various effects these compounds have on different species, individuals, and even tissues; and (iv) how adaptations may evolve in natural populations with continued exposure to EDCs. Several predictions are offered that reflect these new perspectives. Specifically, (i) the threshold assumption will be found not to apply to EDCs because they mimic the actions of endogenous molecules (e.g., estrogen) critical to development; hence, the threshold is automatically exceeded with exposure. (ii) Behavior can compound and magnify the effects of EDCs over successive generations; that is, bioaccumulated EDCs inherited from the mother not only influence the morphological and physiological development of the offspring but also the offsprings' reproductive behavior as adults. This adult behavior, in turn, can have further consequences on the sexual development of their own young. (iii) The sensitivity of a species or an individual to a compound is related to species (individual)-typical concentrations of circulating gonadal steroid hormones. Related to this is the recent finding that alternate forms of the putative receptors are differentially distributed, thereby contributing to the different effects that have been observed. (iv) Except in extraordinary situations, populations often continue to exist in contaminated sites. One possible explanation for this observation that needs to be considered is that animals can rapidly adapt to the nature and level of contamination in their environment. It is unlikely that successive generations coincidentally become insensitive to gonadal steroid hormones fundamentally important as biological regulators of development and reproduction. Rather, adaptive alterations in the genes that encode steroid receptors may occur with chronic exposure to EDCs, allowing the sex hormone receptor to discriminate natural steroids from EDCs. PMID- 11008699 TI - Kin selection and parasite evolution: higher and lower virulence with hard and soft selection. AB - Conventional models predict that low genetic relatedness among parasites that coinfect the same host leads to the evolution of high parasite virulence. Such models assume adaptive responses to hard selection only. We show that if soft selection is allowed to operate, low relatedness leads instead to the evolution of low virulence. With both hard and soft selection, low relatedness increases the conflict among coinfecting parasites. Although parasites can only respond to hard selection by evolving higher virulence and overexploiting their host, they can respond to soft selection by evolving other adaptations, such as interference, that prevent overexploitation. Because interference can entail a cost, the host may actually be underexploited, and virulence will decrease as a result of soft selection. Our analysis also shows that responses to soft selection can have a much stronger effect than responses to hard selection. After hard selection has raised virulence to a level that is an evolutionarily stable strategy, the population, as expected, cannot be invaded by more virulent phenotypes that respond only to hard selection. The population remains susceptible to invasion by a less virulent phenotype that responds to soft selection, however. Thus, hard and soft selection are not just alternatives. Rather, soft selection is expected to prevail and often thwart the evolution of virulence in parasites. We review evidence from several parasite systems and find support for soft selection. Most of the examples involve interference mechanisms that indirectly prevent the evolution of higher virulence. We recognize that hard selection for virulence is more difficult to document, but we take our results to suggest that a kin selection model with soft selection may have general applicability. PMID- 11008700 TI - The diversity of parasites. AB - Parasitism is one of the most successful modes of life displayed by living organisms, as measured by how often it evolved and how many parasitic species are presently in existence. Studying the diversity of parasites is particularly relevant because sympatric diversification may be important in some parasite taxa, and because of the opportunity for independent tests of evolutionary hypotheses in the many separate lineages in which parasitism evolved. Our incomplete knowledge of existing parasite species--the result of a range of phenomena that includes inadequate sampling effort or the lumping of different cryptic species under one name--is not always a major obstacle for the study of parasite diversity. Patterns in the diversity of parasites may be associated with either host or parasite characteristics. The distribution of parasite diversity among host taxa does not simply reflect the species diversity of the host taxa themselves; life history and ecological traits of hosts appear to play important roles. These may determine the likelihood that hosts are colonized by parasite species over evolutionary time. It is not yet clear whether some host traits also favor intrahost speciation and diversification of parasites, and the formation of new parasite species. Certain features of parasites may also be associated with speciation and diversification. Only parasite body size has received much attention; the patterns observed are not greatly different from those of free living species, with small-bodied parasite taxa being more speciose than related large-bodied taxa. Epidemiological parameters such as the basic reproductive rate of parasites, or R0, can also generate predictions regarding the distribution or evolution of parasite diversity. For instance, parasite taxa characterized by high R0 values may be more speciose than related taxa with lower values of R0; such predictions remain untested. Large-scale biogeographical patterns of diversity have only been well studied for metazoan parasites of marine fish; for these parasites, latitudinal patterns can be explained by effects of temperature on speciation rates and epidemiological variables, though other causes are possible. The emphasis for future research must shift from pattern description to the elucidation of the processes responsible for the structure and diversity of parasite faunas. A better integration of ecological and historical (or phylogenetic) approaches to the study of parasite diversity should make this objective possible. PMID- 11008701 TI - Rapid diagnosis of pseudorabies virus infection in swine tissues using the polymerase chain reaction (PCR). AB - In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of suid herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples. PMID- 11008702 TI - [In vitro infection by different strains of Paracoccidioides brasiliensis]. AB - We analyzed the in vitro infection process by P. brasiliensis and the effect of extracellular factor(s) produced on monolayers of mammalian Vero cell lines. The yeast phase of four strains was studied: B339 (avirulent or slightly virulent), U, (intermediate virulence), 93745 and 63265 (both highly virulent). Strains of intermediate and high virulence had higher adherence at first contact (about 16%). Strain B339 had a slower adherence at first contact (8%) than the others during the same period. The production of extracellular proteases, soluble extracellular factor(s) and extracellular antigen gP43 showed no correlation with the in vitro physiopathogenicity of the analyzed strains. We demonstrate that the Vero model presented in this paper is a suitable system to study infection and virulence in vitro. We are currently assessing its usefulness as a tool for the analysis of the interaction between pathogen, host and antifungal agents. PMID- 11008703 TI - Criteria for Candida albicans numerical analysis based on electrophoretic protein patterns. AB - Even in normalized gels, some problems with the choice and position of protein bands always have hindered the processing of electrophoretic data. We have developed a way to establish which criterion best fits the necessities in order to maximize the similarity indexes for numerical analysis. Some repetitions of a Candida albicans strain were carried out in eleven different gels. After staining, the bands were scored in numbers within ranges of +/- n values with increases of one quarter steps (+/- 0.25 kDa, +/- 0.5 kDa, +/- 0.75 kDa, +/- 1.0 kDa, +/- 1.25 kDa) that will limit the possibility of variation for the same bands that will appear on the repetitions. Using this criterion, we have determined that values scored within +/- 1.25 kDa could optimize the minimum limiting value of similarity in different dendrograms built. PMID- 11008704 TI - [Mucocutaneous leishmaniasis in naturally infected dogs in Salta, Argentina]. AB - The objective of the present study is to describe two cases of dogs with mucocutaneous lesions caused by Leishmania spp. Both dogs presented destruction of the nasal septum, hyperemia with soft palate edema and barking alteration due to laryngeal compromise. Biopsies were taken from the lesion border and Leishmania spp. amastigotes were seen in the imprints. The dogs presented positive serology when complex soluble antigen from Leishmania mexicana was used. One of the dogs was also suspected to be infected by Trypanosoma cruzi as suggested by its positive reaction with a purified specific antigen, Ag163B6 cruzipain. Most of the studies concerning leishmaniosis in dogs have described the cutaneous form of this disease in close association with human cases of Leishmania infection instead of the mucocutaneous form described herein. The presence of dogs with mucocutaneous leishmaniosis alerts on an increase of the prevalence of this form in humans, which can cause deforming lesions, alterations of the speech and even an inadequate nutrition due to difficulties in deglutition. PMID- 11008706 TI - [Comparison of 2 techniques for the laboratory diagnosis of influenza virus infections]. AB - The clinical or epidemiological influenza diagnosis require fast, sensitive and accessible techniques for small laboratories. In order to investigate the sensitivity of the methods currently used in Argentina, the rapid diagnosis by indirect immunofluorescent assay (IF) was compared to the rapid viral culture in MDCK cells. The diagnosis of influenza virus infection was performed on 81 nasal and pharyngeal swabs collected from outpatients with upper respiratory infection, influenza-like syndrome. The samples were collected during 1998 winter season and both techniques were tested. The IF specificity and sensitivity obtained were 91.9% and 59.5%, respectively. In the selection of the assay to be used for influenza diagnosis, the limitations of the simplest techniques such as IF should be considered. Furthermore, it is advisable to set up an optimized culture method in complex laboratories since culture is the only technique which allows the reference centers to perform the full characterization of the isolates. PMID- 11008705 TI - Inactivation of Trypanosoma cruzi dihydrolipoamide dehydrogenase by leukocyte myeloperoxidase systems: role of hypochloride and nitrite related radicals. AB - Dihydrolipoamide dehydrogenase (LADH) from Trypanosoma cruzi, the causative agent of Chagas' disease, was inactivated by treatment with myeloperoxidase (MPO) dependent systems. LADH lipoamide reductase and diaphorase activities decreased as a function of incubation time and composition of the MPO/H2O2/halide system, a transient increase preceding the loss of diaphorase activity. Iodide, bromide, thiocyanide and chloride were effective components of MPO/H2O2 or MPO/NADH systems. Catalase prevented LADH inactivation by the MPO/NADH/halide systems in agreement with H2O2 production by NADH-supplemented LADH. Thiol compounds (L cysteine, N-acetylcysteine, penicillamine, N-(2-mercaptopropionylglycine) and Captopril prevented LADH inactivation by the MPO/H2O2/NaCl system and by NaOCl, thus supporting HOCl as agent of the MPO/H2O2/NaCl system. MPO/H2O2/NaNO2 and MPO/NADH/NaNO2 inactivated LADH, the reaction being prevented by MPO inhibitors and thiol compounds. T. cruzi LADH was affected by MPO-dependent systems like myocardial LADH, allowance being made for the variation of the diaphorase activity and the greater sensitivity of the T. cruzi enzyme to MPO/H2O2/halide systems. PMID- 11008707 TI - [Blood cultures: use of presumptive antiobiograms]. AB - Mortality associated to bacteremia varies between 20 and 40% depending upon several factors, such as focus of infection, microorganism, host conditions, etc. It has also been documented that mortality may double when the patient does not receive antibiotic treatment to which the microorganism is susceptible. The objective of our work has been to determine the correlation between disk diffusion antibiogram according to NCCLS guidelines, from isolated colonies, and the one performed directly from the blood culture flask. During 1996, in the Institute of Cardiology and Cardiovascular Surgery (ICYCC) in Buenos Aires City, 81 episodes of bacteremia were studied. In every case, an antibiogram was carried out: 1) from the bottle: a- Directly (D), harvesting 20 microliters in Mueller Hinton agar, b- Diluted (d), previous centrifugation and Gram staining to adjust turbidity equivalent to 0.5 Mc Farland; 2) from isolated colonies, according to NCCLS guidelines. There were almost no major errors, except with two strains of Enterobacter cloacae versus cephalotin. The diluted method was not so convenient to read inhibition zones, especially with staphylococci. With gram-positive bacteria, the main problems appeared in the direct method with erythromycin, oxacillin and ciprofloxacin because of minor errors. With gram-negative bacteria, major errors were observed in the direct method, mainly with piperacillin (7%) and to a lesser extent with piperacillin tazobactam (2%). Except for imipenem, trimethoprim sulfamethoxazoie and cefotaxime, all antimicrobial agents presented minor errors with both methodologies. Based upon the high rate of minor errors, we consider it is important to confirm results obtained with the standard technique (NCCLS), considering as presumptive those results from the blood culture bottles (D and d). PMID- 11008708 TI - [Thermophilic endospores in the environment of a sugar mill in Jujuy]. AB - Twenty six samples from green and scorched sugarcane stems and leaves, sugarmill air dust and raw sugar were analyzed. Thirty nine thermophilic bacilli strains were isolated. Physiological and morphological examinations were carried out according to Bergey's Manual. The strains were identified as B. licheniformis (66.7%), B. coagulans (17.9%), B. stearothermophilus (10.3%) y B. subtilis (5.1%). PMID- 11008709 TI - [Relationship between rhodamine 6G accumulation and fluconazole resistance in Saccharomyces cerevisiae S288c]. AB - Severe mycotic infections are a source of concern in immunocompromised patients or in those who receive chemotherapy for hematological malignant diseases. One of the causes is referred to the appearance of antimycotic resistant microorganisms. Fluconazole is one of the antimycotic used for invasive mycoses treatment. Therefore it is necessary to evaluate the factors that originate this resistance. In the present report the yeast Saccharomyces cerevisiae S288c was used as a model system. In resistant strains the accumulation of the lipophilic cation Rhodamine 6G, L-leucine uptake and growth inhibition by crystal violet dye were determined. The results presented herein demonstrate the correlation between the membrane potential and the resistance to fluconazole presented by S. cerevisiae strain S288c. PMID- 11008711 TI - Membership directory 2000-2001 of the Missouri State Medical Association. PMID- 11008710 TI - [Isolation of Escherichia coli O157:H7 in storm drains in the city of Mar del Plata with bacterial contamination of fecal origin]. AB - The present study was focused on the isolation and characterization of Escherichia coli, particularly the serotype O157:H7, from five combined-sewer outflows waters, which drain into the beaches of Mar del Plata. Seventeen hemolytic uremic syndrome cases were reported in Mar del Plata during the sampling period (May 1995-April 1996) in children between 9 month- and 5 year old, and 3 deaths were recorded. E. coli identification was carried out by biochemical tests. E. coli was detected in 75% of the samples and a total of 98 strains were selected, with 11 sorbol non-fermenting strains. The strains belonged to the O1, O6, O44, O86a, O119 and O168 serogroups. As none of the known virulence factors was detected, strains could not be grouped within any of the diarrheagenic E. coli categories. None of the E. coli strains belonged to the O157:H7 serotype, but E. coli isolation showed fecal contamination in the combined-sewer outflows. Since their waters drain into beaches for recreational use, it is necessary to emphasize the detection of E. coli that would cause severe human illness. Bacterial pollution in combined-sewer outflows draining into Mar del Plata coasts might represent a high risk for human health. PMID- 11008712 TI - [98th Meeting of the German Ophthalmology Society. 21-24 September 2000, Berlin, Germany. Abstracts]. PMID- 11008713 TI - Index 1921-1999. PMID- 11008714 TI - Pennsylvania Medical Society 2000-2001 membership directory. PMID- 11008715 TI - 5th Italian Association of Nuclear Medicine (AIMN) Congress. Pesaro, June 17-20, 2000. Abstracts. PMID- 11008716 TI - [XXVII National Congress of the Spanish Society of Digestive Pathology. La Coruna, 17-20 June 2000. Abstracts]. PMID- 11008717 TI - Microsatellite instability in colorectal adenomas: relevance and clinical importance. AB - A number of investigations have recently been published on the role of microsatellite instability (MSI) in the process of colorectal carcinogenesis. The data concerning colorectal adenomas are difficult to compare (due to differences in the tumor collection, selection and number of analyzed loci, definition of high and low instability and histological types), and this review therefore examines the significance of the results of these publications. We then discuss the extent to which MSI and its effects on the integrity of the genome are early or late events in the malignant transformation, and which clinicopathological features are presented by MSI-positive adenomas. Finally, we consider the clinical importance of microsatellite status for the diagnosis of hereditary nonpolyposis colorectal cancer and the possibility of preventing adenomas and carcinomas by nonsteroidal anti-inflammatory drugs. PMID- 11008718 TI - Increase in the immunostimulatory effect of dendritic cells by pulsing with serum derived from pancreatic and colorectal cancer patients. AB - Both we and others have observed a relative resistance of solid tumor cells to immunological effector cells in vitro, which may be one reason for the clinical phenomenon of resistance of patients with pancreatic carcinoma or other solid tumors to immunological therapeutic approaches. Dendritic cells (DC) are professional antigen-presenting cells which can process and present tumor associated antigens such as CA 19-9. Here we tested DC pulsed with serum containing CA 19-9 for their capacity to stimulate immunological effector cells against pancreatic carcinoma cells. Coculture of immunological effector cells with DC led to a significant increase in cytotoxic activity as measured by a lactic dehydrogenase release assay. Most interestingly, cytotoxic activity against tumor cells was further increased using DC pulsed with patient-derived CA 19-9 containing serum. Similar results have been obtained using either autologous or allogeneic serum from patients with pancreas carcinoma. The effect of serum on the cytotoxicity of effector cells increased in a dose-dependent manner. Interestingly, heat inactivation led to a significant loss of immunostimulatory capacity of the serum. Cytotoxicity was partially inhibited by using an antibody directed against CA 19-9 on the surface of the target cells. Best results were obtained when adding CA 19-9 protein to CA 19-9 containing serum for pulsing of DC. In conclusion, DC pulsed with CA 19-9 containing serum increased the cytotoxic activity of immunological effector cells against pancreatic cancer cells. DC pulsed with CA 19-9 containing serum with or without additional exogenous CA 19-9 protein may have an impact on immunotherapeutic protocols for patients with CA 19-9 secreting tumors. PMID- 11008719 TI - Frequently relapsing Crohn's disease is characterized by persistent elevation in interleukin-6 and soluble interleukin-2 receptor serum levels during remission. AB - We examined immune and inflammatory activation during remission in patients with Crohn's disease who presented with various clinical profiles (prolonged remission vs. relapsing disease). Thirty-six patients with at least 3 years' follow-up starting from a remission period were studied retrospectively. Relapses were defined by a retrospective calculation of the Crohn's disease activity index or by the clinical judgement of the physicians in charge of the patients. Disease course over the study period was assessed by the mean number of annual relapses. Analysis used measurements during remission of the following: erythrocytes sedimentation rate, relative lymphocytosis, acid alpha1-glycoprotein, interleukin 6 (IL-6), and soluble interleukin-2 receptor (sIL-2R) serum levels. During the study period 21 patients experienced at least one relapse and 15 did not. Mean serum levels of sIL-2R and mean relative lymphocytosis in remission significantly discriminated between relapsing and nonrelapsing patients. Only the mean sIL-2R serum level was selected by multivariate analysis, with a cutoff value of 82 pM/1 (sensitivity of 76% and specificity of 80%). The only features correlated with mean number of annual relapses in the relapsing patients were mean serum levels of sIL-2R (r=0.58, P=0.015) and IL-6 in remission (r=0.45, P=0.039). Multivariate analysis demonstrated statistical significance only for the mean serum level of IL-6 (P=0.014). In Crohn's disease the persistent elevation in sIL-2R serum levels during remission corresponds to chronic active disease, while high serum levels of IL-6 in these patients is associated with a high frequency of relapse. PMID- 11008720 TI - Enterovesical fistulas complicating Crohn's disease: clinicopathological features and management. AB - Enterovesical fistula is a relatively rare condition in Crohn's disease. This study was undertaken to examine clinicopathological features and management of enterovesical fistula complicating Crohn's disease. Thirty patients with enterovesical fistula complicating Crohn's disease, treated between 1970 and 1997, were reviewed. Urological symptoms were present in 22 patients; pneumaturia in 18, urinary tract infection in 7, and haematuria in 2. In 5 patients clinical symptoms were successfully managed by conservative treatment, and they required no surgical treatment for enterovesical fistula. Twenty-five patients required surgery. All the patients were treated by resection of diseased bowel and pinching off the dome of the bladder. No patients required resection of the bladder. The Foley catheter was left in situ for an average of 2 weeks after operation. Three patients developed early postoperative complications; two bowel anastomotic leaks, and one intra-abdominal abscess. All these complications were associated with sepsis and multiple fistulas at the time of laparotomy. After a median follow-up of 13 years, 3 patients having postoperative sepsis (anastomotic leak or abscess) developed a recurrent fistula from the ileocolonic anastomosis to the bladder, which required further surgery. In the other 22 patients without postoperative complications there has been no fistula recurrence. In conclusion, the majority of patients with enterovesical fistula required surgical treatment: resection of the diseased bowel and oversewing the defect in the bladder. The fistula recurrence was uncommon, but the presence of sepsis and multiple fistulas at the time of laparotomy increased the incidence of postoperative complications and fistula recurrence. PMID- 11008721 TI - G protein levels and function as an objective measure of depression in patients with functional bowel disorders. AB - Heterotrimeric G proteins play a pivotal role in postreceptor information transduction. These proteins have been implicated in the pathophysiology, diagnosis, and treatment of mood disorders and proposed as a state-dependent biochemical mood marker in mononuclear leukocytes. Irritable bowel syndrome (IBS) is associated with changes in mood, affecting patients' illness perceptions and behavior. We examined whether mononuclear leukocytes of patients with IBS have altered G protein measures. We undertook G protein functional measurements through agonist-enhanced [3H]Gpp(NH)p binding capacity and quantitative measures by immunoblot analysis using anti-Galpha antibodies in mononuclear leukocytes obtained from 19 IBS patients (Rome criteria) and 19 healthy matched subjects. The study groups were similar in age, gender, and years of education. Mononuclear leukocyte functions of G(s) (21.3+/-8.3%) and G(i) (22.2+/-6.7%) proteins in IBS patients were similar to healthy subjects (24.8+/-4.7 and 25.2+/-4.0%, respectively). The relative immunoreactivities of the G(sa) (98.9+/-10.2%) and the G(ia) (104.2+/-11.5%) subunit proteins in mononuclear leukocytes of IBS patients were also similar to those in healthy subjects. Two patients clinically diagnosed as depressed were detected by the G protein assay. The results lend objective support to the contention that major depression is not a causative factor in IBS, nor associated with its severity. The G protein assay may provide an objective biochemical tool for detecting depression in IBS, differentiating it from psychological distress that is commonly diagnosed by subjective tests. PMID- 11008722 TI - Vascular organization in the mesorectum: angiography of rectal resection specimens. AB - The technical advances in rectal cancer surgery are known as the total mesorectal excision. The resection in an anatomically defined plane under direct vision and with sharp dissection distinguishes it conventional rectal surgery. The result must be a complete mesorectum without deep gouges. We performed specimen angiography to confirm completeness of the removed mesorectum. Thirteen total mesorectal excision specimens were examined by angiography after continence preserving resection of rectal carcinoma. In 11 of the 13 cases the vascular supply was exclusively via the superior rectal artery. In two cases with hypoplastic left terminating branches of the superior rectal artery there was additional perfusion via a caudally ascending vessel or via smaller vessels connected laterally. In all specimens both arterial supply and venous outflow were located within the mesorectal fascial sheath. There was no radio-opaque substance leaking from the mesorectal surface in the case of a complete mesorectal specimen. Tiny vascular branches running laterally occurred in 7 of the 13 cases. We found no larger vascular connections branching off in the lateral direction. The rectal blood supply comes almost exclusively through the superior rectal vessels. Thus the fascia covering the mesorectum forms, as far as rectal vascularization is concerned, a closed compartment. The mesorectal vessels are enclosed in the fibrous avascular mesorectal fascia. They run close above the fascia. In the case of an incomplete mesorectal excision the specimen angiography shows a stain leaking from the mesorectal fascia. Our method can be used to confirm the completeness of the removed mesorectum. PMID- 11008723 TI - In vitro peritonitis: basic inflammatory reactions in a two-chamber coculture model of human peritoneum. AB - We developed an in vitro model of the peritoneum by coculturing human umbilical vein endothelial cells (HUVEC) and human peritoneal mesothelial cells (HPMC) to gather information on peritoneal physiology and to closer reflect the in vivo situation in humans. HUVEC and HPMC were seeded on collagen-coated polytetraflourethylene-insert membranes of pore size 3 microm. HUVEC were grown on the bottom of the membrane and HMPC on the top. The confluent cells were monitored by measuring transepithelial resistance and by confocal microscopy. The transmigration of PMNs as an important mechanism during secondary peritonitis was studied in this two-chamber model. PMNs were isolated by density separation. After stimulation of HMPC with the complement factor 5 split product C5a (1 ng/ml) or tumor necrosis factor-alpha (TNF-alpha; 10 or 50 microg/ml) for 1 h, 1 x 10(6) PMN were given to the lower compartment. Controls were cocultured cells without stimulation. After 1, 2, and 6 h nonadherent PMNs in the upper compartment were harvested and counted, interleukin-8 was measured in each compartment, and cells on the membrane were paraffin-embedded for immunohistochemistry. Each experiment was performed four times. Cells grew to confluence within 2-5 days and were detected on their respective seeding side by CD34 and cytokeratin 18 counterstaining. Transmigration of PMNs after C5a or TNF alpha stimulation showed a significant time-dependent increase between 1 h and 6 h (P<0.05). PMNs were found in significantly higher numbers after stimulation with either C5a or TNF-alpha at 1, 2, and 6 h than without stimulation. After stimulation of HPMC, interleukin-8 secretion to the apical compartment increased in a time-dependent fashion, resulting in a gradient between the two chambers. Linear regression analysis revealed significant correlation between transmigrated PMN and interleukin-8 in stimulated cocultures; no correlation was found in controls. This new in vitro peritoneum consisting of cocultured mesothelial and endothelial cells may allow more detailed assessment of peritoneal pathophysiology. Generation of an interleukin-8 gradient affecting the migration of PNMs across the cocultured membrane represents a parameter which may be addressed in further studies. PMID- 11008724 TI - Efficiency of combined colonoscopy and computed tomography for diagnosis of colonic actinomycosis: a retrospective evaluation of eight consecutive patients. AB - The preoperative diagnosis of colonic actinomycosis is accurate in fewer than 20% of patients. Colonoscopy and computed tomography (CT) findings were analyzed to ascertain their diagnostic value and their role in determining therapeutic modality and outcome. Before and after treatment we retrospectively evaluated eight consecutive patients with colonic actinomycosis, all of whom were women with a previous history of intrauterine contraceptive device use. Median follow up period was 20 months (7-57). Localized nodules were found in all cases by colonoscopy; half of these nodules included umbilication at the apex. The colonic mucosa demonstrated normal to moderate inflammation as well as hyperemia and edema. Mucosal ulceration was found in only one case. All cases showed some degree of stenosis. These findings differ from those in other inflammatory or neoplastic diseases of the colon. Abdominopelvic CT revealed extramural involvement of heterogeneous lesions all cases. Contrast enhancement also indicated severe thickening of the colonic wall (7-20 mm) with focal and dense enhancement in six cases. Seven of eight patients required surgery for diagnostic or therapeutic purposes. Complete resolution was determined using both colonoscopy and CT. As more than half of these cases could have been safely managed using an adequate antibiotic treatment, diagnostic studies indicating a high likelihood of colonic actinomycosis should be evaluated to avoid unnecessary surgeries carried out for diagnostic purposes. A combination of colonoscopy and CT appears to be important for both diagnosis and management because of their compensatory findings of mucosal and extramucosal lesions, respectively. PMID- 11008725 TI - A randomized trial of glyceryl trinitrate ointment and nitroglycerin patch in healing of anal fissures. AB - Mean maximum anal resting pressure is directly related to the activity of the smooth muscles of the internal and external sphincters and has been found to be increased in the patients of anal fissure. It has been shown that blood flow at the posterior midline of anoderm is inversely related to the mean maximum anal resting pressure, and topical application of glyceryl trinitrate (GTN) ointments is a very successful treatment. This randomized study was designed to evaluate the relative value of a nitroglycerin patch applied at a distance from the fissure site in healing anal fissure compared to GTN ointment and compared to surgical treatment. Forty-two consecutive patients with chronic anal fissure of more than 4 months' duration were randomized into two equally sized groups: those in group A received 0.2% GTN ointment while those in group B received a 10-mg nitroglycerin patch for 8 weeks. Patients were also asked to rate their pain intensity on a scale of 0-10. Five patients were excluded for various reasons; results were analyzed for the remaining 37 patients (group A, n=18; group B, n=19). A control group C consisted of 12 patients who underwent surgical treatment. Fissures healed completely in 12 of 18 (66.7%) patients in group A, 12 of 19 (63.2%) in group B and 11/12 (91.7%) in group C. The healing rates in groups A and B did not differ significantly (P=0.7), nor was there a difference between these and surgical group C (P=0.13). The local application of GTN ointment and the nitroglycerin patch are both effective, economical, and alternative treatment options for most patients with anal fissures. PMID- 11008726 TI - Glyceryl trinitrate for fissures: patch or paste? PMID- 11008727 TI - Effects of colectomy on bile composition, cholesterol saturation and cholesterol crystal formation in humans. AB - Total or subtotal colectomy is the surgical treatment of choice for patients with ulcerative colitis. Recently it has been reported that colectomy may lead to increased lithogenicity of bile, short nucleation time, cholesterol crystal formation, and gallstone disease. We examined whether colectomy in patients with ulcerative colitis leads to changes in bile composition that predisposes to cholesterol crystal formation and cholesterol gallstone disease. Ten consecutive patients who had previously undergone ileostomy and colectomy because of ulcerative colitis were admitted for ileal pouch surgery. At operation bile was obtained by puncture of the gallbladder. Controls were 35 patients undergoing cholecystectomy (23 for cholesterol gallstone disease and 12 for reasons other than gallstone disease). The gallbladder bile was analyzed for cholesterol crystals, bile acid, and biliary lipid composition, cholesterol saturation, and nucleation time. The colectomized patients had normal biliary lipid composition, normal cholesterol saturation, and normal nucleation time, in contrast to gallstone patients who displayed highly supersaturated bile with a short nucleation time. Thus patients with ileostomy after colectomy because of ulcerative colitis have normal cholesterol saturation and nucleation time of bile. PMID- 11008728 TI - Treatment of acute strangulated internal hemorrhoids by topical application of isosorbide dinitrate ointment. AB - Exogenous nitric oxide has been shown useful in decreasing the internal anal sphincter tone. This study investigated the role of isosorbide dinitrate in the treatment of patients with acute strangulated internal hemorrhoids, thereby avoiding the risk of continence disturbances following conventional surgical treatment. Four male patients (median age 35 years, range 30-42) with acute strangulated hemorrhoids were treated with 1% isosorbide dinitrate. The ointment was applied to the anoderm. This application was repeated every 3 h during daytime for 2 weeks. Significant pain relief was achieved within 1 day, while transient mild headache was experienced during the first 2 days. Within 1 week the hemorrhoids became reducible. Thereafter the hemorrhoids could be treated adequately by rubber band ligation. The alternative treatment of patients with acute strangulation of prolapsed internal hemorrhoids is effective. This nonsurgical, i.e., reversible reduction of sphincter tone is an attractive alternative. PMID- 11008729 TI - Congenital coxa vara: computed tomographic analysis of femoral retroversion and the triangular metaphyseal fragment. AB - Three patients with congenital coxa vara studied with two- and three-dimensional computed tomographic (2DCT and 3DCT) methods are reported. In all cases, the femoral retroversion was documented and subsequently corrected by proximal femoral osteotomy. In two patients with isolated coxa vara, the physeal-femoral neck angle was decreased as seen in slipped capital femoral epiphysis in adolescents. Our studies suggest that the triangular metaphyseal fragment reflects a Salter-Harris type II separation pattern through the defective femoral neck. The epiphysis and attached triangular fragment slip from the normal superoanterior portion of the neck in an inferior-posterior direction. The treating surgeon should be aware of the often marked femoral retroversion component present in severe congenital coxa vara. This knowledge allows surgical planning for corrective osteotomies that will better normalize hip mechanics. A combination of marked valgus and flexion with internal rotation of the distal fragment are required to fully correct the deformity. PMID- 11008730 TI - Classification of metaphyseal change with magnetic resonance imaging in Legg Calve-Perthes disease. AB - Seventy-eight patients (85 affected hips and 71 unaffected hips) with Legg-Calve Perthes disease were included in this study to evaluate the metaphyseal change in radiographs and magnetic resonance imaging (MRI) and to define the type of the metaphyseal cyst according to presence or absence of the epiphyseal involvement. The content of the metaphyseal cyst was evaluated by using T1,T2, proton, and gadolinium-enhanced T1-weighted MRI scans. Among 85 hips, there were no changes in 32 hips, marrow edema in 13 hips, false cyst with epiphyseal involvement in 28 hips, and true cyst without epiphyseal involvement in 12 hips. Granulation tissue was found in the false cysts and water-rich fibrotic tissue was found in the true cysts based on the MRI scans. The metaphyseal change in MRI scans was shown in 71% of groups 3 and 4 and in 35% of groups 1 and 2 according to the Catterall classification, and 52% of group A, 56% of group B, and 86% of group C according to the Herring classification. Of the 30 hips at the avascular stage, 33% showed metaphyseal cyst in MRI scans. Of the 53 hips at the fragmentation stage, 60% showed the metaphyseal cyst. PMID- 11008731 TI - The effect of pediatric orthopaedic experience on interobserver and intraobserver reliability of the herring lateral pillar classification of Perthes disease. AB - An analysis was done of the effect of surgeons' pediatric orthopaedic experience on the classification of Perthes disease according to the lateral pillar classification described by Herring. Five observers with varied pediatric orthopaedic experience reviewed anteroposterior (AP) pelvis radiographs of 33 patients in the fragmentation phase of Perthes disease and classified each case on three separate occasions at least 24 hours apart. Frog-leg lateral-view radiographs taken at the same time were also classified using the same criteria based on the femoral head anterior column. Kappa statistics showed good agreement for intra- and interobserver reliability of classification for both AP and frog lateral radiographs. There were no statistically significant variations among the reviewers. Twenty-four percent of the cases had a classification one grade worse on the lateral radiograph compared to the AP view. The lateral pillar classification provides a reproducible radiographic technique for Perthes disease characterization independent of pediatric orthopaedic experience. PMID- 11008732 TI - Radiographic results of proximal femoral varus osteotomy in Legg-Calve-Perthes disease. AB - A retrospective review of 116 Legg-Calve-Perthes patients with varus osteotomy for Herring group B and C disease involvement was conducted. Preoperative assessment consisting of anteroposterior pelvis and frog leg lateral radiographs and an arthrogram was performed on all patients. Radiographic extent of disease was classified using Herring's lateral pillar classification system. Radiographic outcome was assessed utilizing Stulberg's classification to grade residual deformity. Follow-up averaged 6 years 9 months (range, 3 years to 15 years 3 months). Stulberg class I and II (spherically congruent) results were obtained in 86% of patients younger than 9 years old with Herring class B disease. Patients 9 years and older with Herring class B disease had spherically congruent results in 67% of cases. Patients with Herring class C disease 9 years and older had spherically congruent results in 30% of cases and 43% spherically congruent results in those younger than 9 years. PMID- 11008733 TI - Limited magnetic resonance imaging examination after surgical reduction of developmental dysplasia of the hip. AB - We evaluated the utility of limited magnetic resonance imaging (MRI) after surgical reduction and spica casting for developmental dysplasia of the hip (DDH) with respect to feasibility, time, need for sedation, and charge. Eighteen limited MRI examinations were performed in 10 children (mean age, 9 months) with a 1.5-T magnet. Twenty-eight dysplastic hips were imaged within 4 hours of surgery. Mean imaging time for two sequences was approximately 3 minutes. All examinations were interpretable, although three were degraded by motion. No child required sedation, additional sequences, or repeat study. Two thirds of the dysplastic hips had no proximal femoral ossification center on MRI. A reduced charge was assigned to the examination. We conclude that using limited MRI to confirm intraoperative reduction for DDH is feasible and desirable. The lack of radiation or need for sedation, the reduced charge that reflects the short time needed for imaging, and the superb visualization of nonosseous structures have made it our preferred method to evaluate surgical reduction of DDH. PMID- 11008734 TI - Intraoperative instability for developmental dysplasia of the hip in children 12 to 18 months of age as a guide to Salter osteotomy. AB - There is still debate on the necessity of Salter osteotomy for developmental dysplasia of hip (DDH) between 12 and 18 months of age. The goals of this study were to investigate the correlating factors of intraoperative instability as a guide to the additional Salter osteotomy and to evaluate the radiographic and clinical results. Stability could not be achieved in 63% of 84 hips with soft tissue releases alone. The existence of three pathologic findings (grade of dislocation, inverted labrum, and excessive anteversion) and absence of two surgical procedures (transverse acetabular ligament incision and iliopsoas osteotomy) significantly correlates with instability. At follow-up, the acetabular remodeling with or without Salter osteotomy was similar. We conclude that the Salter osteotomy does not interfere with the acetabular remodeling and has no major disadvantages for children at that age but can help to improve the stability of the hip. PMID- 11008735 TI - Preliminary traction as a single determinant of avascular necrosis in developmental dislocation of the hip. AB - The role of preliminary traction before closed reduction in the treatment of developmental dysplasia of the hip has been questioned by many authors lately. However, the studies advocating or opposing the use of this treatment modality include several other parameters besides traction. Thus, it is unclear whether the affection is the result of preliminary traction or concurrent variables such as the "human position." This study aimed to put forward the effect of preliminary traction as a single determinant of avascular necrosis. We had two groups of patients who had developmental dislocation of the hip. The first group consisted of 52 patients treated with preliminary traction before closed reduction and the other group comprised 40 patients treated with closed reduction without preliminary traction. Both groups were similar in age, gender, side, level of dislocation, and method and duration of immobilization. Three patients from the first group developed avascular necrosis, which was not statistically significant. Preliminary traction did not affect the rate of avascular necrosis. PMID- 11008736 TI - Total hip arthroplasty after childhood septic hip in patients younger than 25 years of age. AB - Childhood septic hip should usually be treated immediately by arthrotomy and antibiotic. Even if treated correctly, the affected hip may become osteoarthritic and functionally disabling. Usually the literature is not in favor of total hip arthroplasty in young patients, and the reports are on patients older than 32 years of age. We present here a unique group of very young patients with early coxarthrosis caused by septic hip in childhood, with an average age of 19.14 years (range, 14-25) at the time of the arthroplasty. The Harris hip score improved from a preoperative mean of 58.43 to a postoperative mean of 94.14. The follow-up period ranged between 2 and 24 years, with an average of 8.14 years. We conclude that total hip arthroplasty in young people with early coxarthrosis caused by septic hip in childhood provides good functional results. PMID- 11008737 TI - Ultrasonographic evaluation of hip morphology in osteochondrodysplasias. AB - The developing hip in children with osteochondrodysplasias has not been well described because of delayed ossification and limitations of conventional radiologic techniques. Twenty-four children with various osteochondrodysplasias were evaluated by ultrasonography. Variation in the configuration of the acetabulum included a horizontal acetabular roof owing to delayed iliac development and a notched acetabular roof with lateral bone deficiency. All children had thickened acetabular cartilage except for one child with osteogenesis imperfecta. Coxa vara was a common finding. All neonates displayed a very small beta angle (mean, 42 degrees) because the labrum lay more vertically, secondary to deep engagement of the femoral head in the acetabulum. Proximal femoral ossification was delayed in most children, which allows use of ultrasonography at a later age than is possible in the normal pediatric population. Hip ultrasonography in children with skeletal dysplasias can aid in early diagnosis and is useful in assessing hip morphology and development. PMID- 11008738 TI - The genetic basis of the osteochondrodysplasias. AB - The osteochondrodysplasias are a heterogeneous group of disorders characterized by abnormal growth and remodeling of cartilage and bone, affecting from 2 to 4.7 per 10,000 individuals. Most osteochondrodysplasias are heritable and many have elaborate patterns of genetic transmission. Affected individuals generally require management by multidisciplinary teams of specialists. In this review, we divide the osteochondrodysplasias into groups based on their genetic relationships, including mutations in various types of collagen, fibroblast growth factor, cartilage oligomeric matrix protein, parathyroid hormone receptor, the diastrophic dysplasia sulfate transporter, enzymes such as steroid sulfatases, transcription factor SOX9, and a cysteine proteinase, cathepsin K. We describe the major osteochondrodysplasias, define their causes and clinical manifestations, and provide the orthopaedic surgeon with an understanding of the underlying molecular defects as well as the anatomical aspects of these disorders. PMID- 11008739 TI - Treatment of late-onset tibia vara using afghan percutaneous osteotomy and orthofix external fixation. AB - The Afghan osteotomy was combined with external fixation on 19 patients (23 extremities) with late-onset tibia vara. The average weight was 258 lb and all patients weighed >95th percentile. The average preoperative deformity was 28.2 degrees. The average intraoperative correction was 27.6 degrees. Average healing time was 141 days. The mean follow-up was 2.7 years. Based on radiographic correction, at long-term follow-up, there were 15 excellent, two fair, and six poor results. The quality of the initial correction was the only significant variable, and it was borderline (p = 0.0587). Complications included loss of alignment, peroneal nerve palsy, superficial pin tract infection, deep infection, and fracture. This method offers a technically simple procedure with a relatively low complication rate. It allows early mobilization and provides the ability to manipulate the correction postoperatively. An excellent long-term outcome is predicated on achieving an acceptable initial correction as determined by intraoperative mechanical axis radiographs. PMID- 11008740 TI - Early characteristic findings in bowleg deformities: evaluation using magnetic resonance imaging. AB - We used magnetic resonance imaging (MRI) to evaluate bowleg deformities in infancy. Twenty-five tibiae of 13 infants were examined and divided into two groups based on MRI findings: group A had high intensity area in the medial epiphyseal cartilage on T2-weighted images. Group B had depression of medial physis and abnormal signal in the perichondrial region in addition to the epiphyseal lesion. At the final follow-up, all cases in group A demonstrated normal lower leg alignments, whereas five cases in group B showed characteristic roentogenographic findings of Blount's disease. The improvement rate of metaphyseal-diaphyseal angle was correlated with this classification. These findings suggested that abnormal findings in physis and perichondrial region might be preliminary findings in early stage of Blount's disease. The high intensity areas in the medial epiphyseal cartilage were commonly found among the cases with bowing deformities, which suggested that there might be a common pathomechanism between physiologic bowing and infantile Blount's disease. PMID- 11008741 TI - Tibial hemimelia of a different class. AB - A variant of tibial hemimelia, previously undescribed in the literature and not classifiable by either of the established classification systems, is described. The features of this condition include a short, deformed tibia; proximal subluxation of the fibula at the knee; a normal knee joint and an ankle joint that may look abnormal but falls short of true diastasis of the joint. Treatment of two cases by differential lengthening of the tibia and fibula using the Ilizarov device is described. This form of tibial hemimelia should be recognised as a separate variant as preservation of the foot and ankle with expected excellent function should be possible, unlike the more severe forms of this condition. PMID- 11008742 TI - Internal derotation osteotomy of the tibia: pre- and postoperative gait analysis in persons with high sacral myelomeningocele. AB - Rotational deformities of the lower extremities are common in patients with myelomeningocele. In these situations, surgical correction is often necessary. We conducted a retrospective review of eight ambulatory patients with high sacral myelomeningocele and external tibial torsion who underwent 10 distal tibia and fibular internal derotation osteotomies. All patients had an increased valgus knee stress preoperatively. Pre- and postoperative three-dimensional gait analysis was used to evaluate coronal plane knee moments and dynamic sagittal plane knee motion. Postoperatively, a significant improvement in the abnormal internal knee varus moment (p < 0.005) as well as a significant increase in the stance phase knee extension (p < 0.01) was seen. Three patients had resolution of preoperative knee pain. We believe that patients with increased knee stress secondary to excessive external tibial torsion will benefit from a tibial derotation osteotomy that could delay or prevent the onset of late degenerative changes about the knee. PMID- 11008743 TI - An examination of the knee function during gait in children with myelomeningocele. AB - In this retrospective study, 37 patients with myelomeningocele who had undergone gait analysis were examined. Patients were divided into groups based on the level of involvement (29 sides: L4; 26 sides: L5; 19 sides: S1-2). Results showed increased knee flexion and associated knee extensor moments with increasing level of neurologic involvement. The mean coronal plane knee position in stance was normal in all groups and not related to coronal plane knee moment. However, there was an increased incidence of a net knee adductor moment in stance with increasing involvement (mean, 0.02 +/-0.18 N.m/kg for the L4 group). The presence of a visual valgus thrust based on video records was not reliable in predicting an abnormal knee coronal plane moment. An abnormal knee adductor moment in stance was most highly related to coronal plane trunk motion (r = -0.62) and not tibial torsion (r = -0.340). Increased transverse plane range of motion of the knee was most highly related to transverse plane trunk motion (r = 0.67). PMID- 11008744 TI - Radiographic abnormalities and clinical symptoms associated with patella alta in ambulatory children with cerebral palsy. AB - One hundred seventeen randomly selected, skeletally immature, ambulatory children with spastic cerebral palsy were evaluated with clinical examination and lateral radiographs of the knee. Radiographic abnormalities within either the patella or the proximal tibia were present in 21% (41 knees) of the 193 affected knees. The most common radiographic findings were elevation of the tibial tubercle in 34 knees and fragmentation of one pole of the patella in seven knees. Clinical symptoms including pain, swelling, and tenderness were present in 32% of those children with radiographic findings (13 of 41 knees). Patella alta was seen in nearly all children in the study. The radiographic findings appeared to be chronic, were usually asymptomatic, and have required no treatment to date. PMID- 11008745 TI - Broomstick plaster with a removable abduction bar: a simple technique that facilitates early postoperative physical therapy and handling. AB - We present a simple technique utilising a broomstick plaster with a removable abduction bar. This facilitates early postoperative physical therapy and handling after surgery for lower limb problems in patients with cerebral palsy. PMID- 11008746 TI - Evaluation of the walking pattern in clubfoot patients who received early intensive treatment. AB - The walking pattern in a group of nine adult male subjects who had received early intensive treatment for congenital clubfoot was evaluated and compared to the walking pattern in a control group of 15 adult healthy male subjects. All subjects were filmed with a five-camera video system as they walked across two force plates. A three-dimensional inverse dynamics approach was used to calculate average joint angles, moments, power, and work. The results showed that there were no differences in the joint angles between the two groups. The overall patterns of the joint moments were very similar in the two groups. However, analysis revealed a smaller ankle joint moment and larger knee and hip joint moments in those with clubfoot. It was concluded that the larger knee and hip joint moments served as compensation for the smaller ankle moment. The reduced ankle moment and work developed about the ankle joint in the clubfeet could possibly be owing to weaker plantar flexors. In conclusion, gait analysis can be an important tool when evaluating treatment for clubfoot. However, further investigation is needed to determine whether the higher hip and knee joint moments observed in subjects with clubfoot may lead to the development of knee or hip joint pathologies. PMID- 11008747 TI - Comparison of the soft-tissue release methods in idiopathic clubfoot. AB - The aim of this study was to compare the clinical and radiologic results of three different surgical procedures (posterior release with lengthening of the tendo calcaneus and posterior capsulotomy, Turco's technique of posteromedial release, and Simons' technique of complete subtalar release) in idiopathic clubfoot in 77 patients who were operated on at 10 months of age or younger. The clinical examination alone is not sufficient to determine the type of the surgical intervention needed in idiopathic clubfoot; the decision must be supported by the radiologic parameters (anteroposterior and lateral talocalcaneal, first metatarsal-talar, and lateral tibiotalar angles) and all the components of the multiplanar deformity must be corrected at the same time. Simons' technique of complete subtalar release was found to be the most efficient method of surgery both functionally and radiologically in cases of idiopathic clubfoot in infants. The overcorrection of the deformity as a consequence of the concern that the tarsal alignment did not improve adequately is a mistake we make not infrequently and in our opinion this may be avoided by taking perioperative radiographs. PMID- 11008748 TI - Talectomy for clubfoot in arthrogryposis. AB - We reviewed the results of 101 talectomies in 56 patients with arthrogrypotic clubfeet. The average age at the time of surgery was 4.3 years and the mean follow-up was 6 years. Talectomy was performed as a primary procedure in 16 feet and as a salvage procedure in 85 feet that underwent other surgical procedures before talectomy. We graded our results as good when the foot was plantigrade, able to wear regular shoes, pain free, and, very important, patient satisfaction. We used a chi2 statistical test and, after comparing results with age <4 to >4 years at time of surgery, tendo Achilles tenotomy, time of casting, radiological complete excision of talus, and transcalcaneal pin placement, only the immobilization time needed to be statistically significant to achieve a good result. We conclude that feet must be individualized for treatment and that, after reduction of the calcaneus in the mortise, a short leg cast must be placed for 8 weeks to maintain position and alignment. PMID- 11008749 TI - Scoliosis in arthrogryposis multiplex congenita: results after nonsurgical and surgical treatment. AB - Forty-six patients (25 male and 21 female) with arthrogryposis multiplex congenita who were seen at the Alfred I. duPont Hospital for Children between the years 1940 and 1997 were evaluated to assess the prevalence and patterns of scoliosis and the long-term results after both nonsurgical and surgical treatment methods. We found the prevalence of scoliosis to be 65.9% (32 of 46 patients). A single thoracolumbar curve was the predominant curve pattern. No congenital curve types or vertebral anomalies were seen in our group of patients. In the nonsurgical group, the mean curve was <30 degrees at follow-up. In the surgical group, the mean primary curve before spinal arthrodesis was 78.5 degrees. Three patients in the surgical group who were nonambulators have become household ambulators at the most recent follow-up. We recommend bracing in patients who are ambulators and have a curve of <30 degrees. Combined anterior and posterior spinal arthrodesis gave the best results. PMID- 11008750 TI - The induction of congenital spinal deformities in mice by maternal carbon monoxide exposure. AB - Carbon monoxide (CO) has been shown to be teratogenic in mice. High altitude hypoxia has also been shown to induce congenital vertebral anomalies in mice. The purpose of this study was to investigate the effect of maternal hypoxia owing to CO exposure and the production of congenital spinal deformities in the offspring. Sixty DBA-1J mice were bred using polygamous timed breeding methods. Pregnant females were exposed to 200, 400, or 600 ppm CO using a custom-designed gas blender system. Seven-hour exposures were performed on day 8.5, 9.5, or 10.5 of the 21-day gestation cycle. The neonates were euthanized at birth; the specimens were fixed, eviscerated, and radiographed. Congenital spinal deformities were observed (wedge, hemi, fused, and missing vertebrae; fused ribs) and were located in all regions of the spine. There was a statistically significant difference in the number of spinal deformities between all groups, with no defects in the controls and a 77% incidence at 600 ppm (p < 0.0001). There was no apparent correlation between the time of exposure and defect location. The most sensitive time of gestation was 9.5 days. We identified an animal model of congenital spinal deformities that compares favorably with the evidence of human congenital spinal deformities in cases of maternal exposure to CO and other gas and chemical fumes. PMID- 11008751 TI - Analysis of the STIF technique for spino-pelvic fixation: clinical results in 19 patients with neuromuscular scoliosis. AB - The STIF (spinopelvic transiliac fixation) technique for lumbosacral fusion was developed by the authors as an alternative to the Luque-Galveston technique. The results observed in this initial series of 27 neuromuscular scoliosis patients treated with the STIF technique are reported. With a minimum of 24 months of radiographic follow-up in 19 patients, the percentage of correction of scoliosis curvature and pelvic obliquity was superior to that reported in the literature. The rates of complications and pseudarthrosis in this series are typical for this patient population. The STIF technique facilitates compression across the sacroiliac joints, which promotes sacroiliac joint fusion and can provide a stable base for curvature correction and lumbosacral fusion. Despite the severe coronal and sagittal plane curves in this group of patients, total operative time also compares favorably to that reported in the literature. The STIF technique requires a well-developed posterior iliac apophysis, which may not be present in younger pediatric patients. PMID- 11008752 TI - Cervical spondylolysis in children: is it posttraumatic? AB - Cervical spondylolysis is a rare defect of unknown etiology. Five cases of cervical spondylolysis as well as two cases of fractures of the pedicles of C2 in infants are presented. Comparison of the cases suggests that a fracture at birth or in infancy may be the cause of some cases of cervical spondylolysis. PMID- 11008754 TI - Relationship of factors affecting age of onset of independent ambulation. AB - Despite the standard available pediatric developmental scales and popular lore that girls walk at an earlier age than boys, no large-scale evaluation of the age of onset of independent ambulation has been previously published. The purpose of this study was the prospective epidemiologic evaluation of a large heterogeneous group of normal children to determine the effect of gender, race, birth order, and socioeconomic status on the age of onset of independent ambulation. The study cohort consisted of 986 children (575 male, 471 female). A multivariable analysis of covariance model was used to examine the effects of race, gender, income, and birth order on age at ambulation. After controlling for the other variables in the model, race was the only statistically significant predictor of age at ambulation (p < 0.0001), with black children walking at a younger age (10.9 +/- 2.1 months) than white children (11.6 +/- 2.3 months). Overall, the independent variables included in the model were only able to explain 2.5% of the variance of age at ambulation. PMID- 11008753 TI - Retrospective review of osteoarticular infections in a pediatric sickle cell age group. AB - Patients with sickle cell disease have been documented to be particularly susceptible to osteoarticular infections. Controversy exists concerning the bacteriology, etiology, and clinical presentation in differentiating osteoarticular infections from bone infarct. We retrospectively reviewed all cases from our institution over the past 22 years of osteoarticular infections in children who carry the diagnosis of sickle cell disease. Two thousand consecutive patient charts of children enrolled in the Pediatric Sickle Cell Clinic of our institution between 1973 and 1995 were evaluated. There were 14 cases of bone or joint infections (10 osteomyelitis, four septic arthritis). There was one case of multicentric osteomyelitis and one case of meningitis complicating the septic arthritis. There were nine male and five female patients with ages ranging from 6 months to 17 years (mean, 8.0). All patients were noted to have hemoglobin SS. The predominant presenting symptoms were pain (79% of cases) and swelling (71% of cases). The most frequent physical findings were fever >38.2 degrees C (71% of cases) and tenderness (86% of cases). Ninety-three percent of the children had a white blood count exceeding 15,000/mm3 (range, 7,900-32,300). Westergren sedimentation rates ranged from 14 to 89 mm/h with 93% of the children exceeding the normal value in our hospital. Cultures were positive in 75% of tissue biopsies, 58% of the blood cultures, and 70% of the bone or joint aspirates. The most common offending organism found in osteomyelitis was Salmonella (eight of 10 cases); however, no predominant organism found was identified in cases of septic arthritis. Radiographs and bone scans were of limited value in the differential diagnosis between osteoarticular infections and bone infarction. Early diagnosis and treatment of osteoarticular infections is key to satisfactory outcome. This study suggests that an ill-appearing patient with a fever >38.2 degrees C, pain, and swelling should prompt the physician to aspirate or biopsy the area and not rely on diagnostic studies that we found to be unreliable. PMID- 11008755 TI - Spontaneous regression of osteochondromatosis of the radius after lengthening of the ulna: a case report. AB - Despite the high incidence, the natural biologic behavior of the osteochondroma is not yet known. In particular, the spontaneous regression of the exostosis is very rare and as yet an undefined phenomenon. Since the first description by Hunter in 1835, only 11 cases have been reported. On reviewing the reports in the literature, all cases were regressed spontaneously without significant mechanical stimuli such as surgical procedures. The reasons for the spontaneous regression were not explained sufficiently. This study describes one additional case of the spontaneous regression of osteochondromatosis of the radius and ulna after change of the mechanical strain or stress with lengthening of the ulna in a patient with multiple hereditary exostosis. The radiographic findings in this study are presented with some hypotheses explaining this interesting phenomenon. PMID- 11008756 TI - Abnormalities in the uninvolved lower limb in children with spastic hemiplegia: the effect of actual and functional leg-length discrepancy. PMID- 11008757 TI - A selective optical sensor based on [9]mercuracarborand-3, a new type of ionophore with a chloride complexing cavity. AB - A highly selective optical sensor for chloride, based on the multidentate Lewis acid ionophore [9]mercuracarborand-3, is described herein. This sensor is constructed by embedding the mercuracarborand ionophore, a suitable pH-sensitive lipophilic dye, and lipophilic cationic sites in a plasticized polymeric membrane. The multiple complementary interactions offered by the preorganized complexing cavity of [9]mercuracarborand-3 is shown to control the anion selectivity pattern of the optical film. The film exhibits a significantly enhanced selectivity for chloride over a variety of lipophilic anions such as perchlorate, nitrate, salicylate, and thiocyanate. Furthermore, the optical selectivity coefficients obtained for chloride over other biologically relevant anions are shown to meet the selectivity requirements for the determination of chloride in physiological fluids, unlike previously reported chloride optical sensors. In addition, the optical film responds to chloride reversibly over a wide dynamic range (16 microM-136 mM) with fast response and recovery times. PMID- 11008758 TI - Modeling interparticle and intraparticle (perfusive) electroosmotic flow in capillary electrochromatography AB - A model to estimate the extent of intraparticle, or perfusive, electroosmotic flow (EOF) in CEC capillaries packed with macroporous particles has been developed. Nucleosil packings (d(p) = 7 microm) having nominal pore sizes of 500, 1000, and 4000 A were studied. Intraparticle pores ranging from 50 to 10000 A in diameter were partitioned into 995 intervals of 10 A. Using pore size distribution data for the sorbents obtained by mercury intrusion porosimetry, fractions of the total column void volume contributed by pores in the range of interest were determined. The average channel diameter of the interstitial space was estimated from the d(p) of the packing; its fraction of total column volume was determined from the interstitial porosity. Estimations of relative EOF velocity in the intraparticle and interstitial channels were made by treating the channels as parallel cylindrical capillary tubes. Relative EOF values were combined with the volume fraction data and used as weighting factors in calculating an effective particle diameter (d(p,eff)) for each set of conditions (i.e., packing type, ionic strength of eluent). Values of d(p,eff) generated by the model correctly predict the trends observed in the experimental data. At the lowest ionic strength, plate height correlated inversely with the pore size of packing (h4000 A < h1000 A < h500 A). Rate curves for each column tended toward lower plate heights with increasing eluent ionic strength before converging at some limiting point. The point of convergence was reached at moderate ionic strengths for the larger pore media (1000 and 4000 A) and higher ionic strength for the 500 A. PMID- 11008759 TI - Subfemtomole MS and MS/MS peptide sequence analysis using nano-HPLC micro-ESI fourier transform ion cyclotron resonance mass spectrometry. AB - Subfemtomole peptide sequence analysis has been achieved using microcapillary HPLC columns, with integrated nanoelectrospray emitters, coupled directly to a Fourier transform ion cyclotron resonance mass spectrometer. Accurate mass (+/ 0.010 Da) peptide maps are generated from a standard six-protein digest mixture, whose principle components span a concentration dynamic range of 1000:1. Iterative searches against approximately 189000 entries in the OWL database readily identify each protein, with high sequence coverage (20-60%), from as little as 10 amol loaded on-column. In addition, a simple variable-flow HPLC apparatus provides for on-line tandem mass spectrometric analysis of tryptic peptides at the 400-amol level. MS/MS data are searched against approximately 280000 entries in a nonredundant protein database using SEQUEST. Accurate precursor and product ion mass information readily identifies primary amino acid sequences differing by asparagine vs aspartic acid (deltam = 0.98 Da) and glutamine vs lysine (deltam = 0.036 Da). PMID- 11008761 TI - Multichannel mode-filtered light detection based on an optical fiber for small volume chemical analysis AB - A novel mode-filtered light detection method is described in which an unjacketed optical fiber is inserted into a transparent capillary tube and three or more detection channels are set on the capillary side for different distances from the port of the fiber. This new method is the basis of synchronization of separation and analysis, with which a modern multidimensional analysis apparatus will be constructed. For samples of different concentration, the more close to the laser incidence port of the fiber the detector has been set up, the greater the change of the intensity profile of mode-filtered light (deltaI(F)) that is obtained. Vice versa, if a parameter alpha = I0/In is established and, instead of a mode filtered light signal, the reversed alteration trend of deltaalpha is found in comparison with the alteration trend of deltaI(F), the reason is that the background rapidly lowers with the increasing distance to the laser incidence port of the fiber; moreover, the mode-filtered light signal decreases slowly with it. With the present method and apparatus, glucose and glycerol have been determined, with good reproducibility and stability and a small sample volume. Furthermore, a real sample of glucose injection is measured for a detection volume of 5 microL, and an acceptable result is observed. PMID- 11008760 TI - A mass spectrometry method for the determination of enantiomeric excess in mixtures of D,L-amino acids. AB - A mass spectrometry method based on ion/molecule reactions for determining enantiomeric excess in mixtures of amino acids is illustrated. Protonated complexes of amino acids with beta-cyclodextrin are produced in the gas phase using electrospray ionization. Under a background pressure of N-propylamine, the amino acid is replaced by the alkylamine. The rates of the guest exchange reaction vary with the chirality of the amino acid. A calibration curve employing varying mixtures of D- and L-amino acids is produced and the fractions of the D isomer in the test mixtures are determined. The quality of the curve is compared to the chiral selectivity of the reaction. In general, the best calibration curves are obtained in systems with the greatest selectivity. PMID- 11008762 TI - Detection and quantification of metals in organic materials by laser-SNMS with nonresonant multiphoton ionization. AB - We have shown that the sensitive detection and in favorable cases the quantification of metals in organic materials by laser-SNMS with nonresonant multiphoton ionization (NRMPI) is possible. As a model system, sputter-deposited submonolayer coverages of metals on polymer surfaces (polycarbonate, poly(vinylidene chloride), polyimide) were investigated. By use of these samples, relative sensitivity factors and detection limits of several metals (Be, Cr, Mn, Fe, Co, Ni, Mo, W) were determined using laser-SNMS with NRMPI. The relative sensitivity factors for this kind of sample show a high level of agreement with those for metals sputtered from alloys. The detection limits ( 1 ppm of a monolayer) are almost the same as for inorganic matrixes such as Si or GaAs. Laser-SNMS with NRMPI was also used for the determination of the elemental composition of the active centers of metalloproteins (namely, the purple acid phosphatases extracted from sweet potatoes and from red kidney beans). These results have shown the ability of laser-SNMS to detect metal atoms bound to organic macromolecules with an atom concentration as low as 1 ppm. In comparison to TOF-SIMS, laser-SNMS is more sensitive for metal detection in organic matrixes, since the secondary ion yields observed for these matrixes are reduced compared to matrixes optimized for high secondary ion emission, such as, for example, highly oxidized surfaces. PMID- 11008763 TI - An assay for ascorbic acid based on polyaniline-coated microplates. AB - A technique for modification of the microtiter reader plates well with a polyaniline (PANI) film sensitive for ascorbic acid is presented. The principle of the analyte detection is based on monitoring the changes in optical absorption of the PANI film resulting from the reduction process initiated by ascorbic acid. The detection limit for ascorbic acid is 1 mg/L. Testing with real samples (soft drinks, fruit juices) gave good correlation of the method with iodimetric titration. High sensitivity, stability, and good reproducibility of the measurements make the proposed system an attractive alternative to traditional assays, used in medicine, ecology, and biotechnology. PMID- 11008764 TI - Fabrication, characterization, and enzymatic activity of encapsulated fungal protease--fatty lipid biocomposite films. AB - Encapsulation of an aspartic protease from the fungus Aspergillus saitoi (F-prot) in thermally evaporated fatty acid films by a simple beaker-based immersion technique under enzyme-friendly conditions is described. The approach is based on diffusion of the enzyme from aqueous solution, driven primarily by attractive electrostatic interaction between charged groups on the enzyme surface and ionized lipid molecules in the film. The encapsulated enzyme molecules could be "pumped out" of the biocomposite film into solution by modulating the electrostatic interaction between the enzyme and fatty acid molecules via solution pH variation. The kinetics of F-prot diffusion into the acid films was followed using quartz crystal microgravimetry measurements while the secondary and tertiary structure of the enzyme in the lipid matrix was studied using Fourier transform infrared (FT-IR) and fluorescence spectroscopies. FT-IR and fluorescence measurements indicated little perturbation to the native structure of the enzyme. A chemical analysis of the F-prot-fatty acid biocomposite film was also performed using X-ray photoelectron spectroscopy. The encapsulated F-prot molecules showed catalytic activity (as estimated by reaction with hemoglobin) comparable to free enzyme molecules in solution, indicating facile access of biological analytes/reactants in solution to the enzyme molecules. The advantages/disadvantages of this approach vis-a-vis methods currently used for encapsulation of biomolecules are briefly discussed. PMID- 11008765 TI - Multielement analysis of polyethylene using solid sampling electrothermal vaporization ICP mass spectrometry AB - Next to laser ablation (LA) also electrothermal vaporization (ETV) from a graphite furnace as a means of sample introduction opens possibilities for direct analysis of solid samples using inductively coupled plasma mass spectrometry (ICPMS). In this paper, it is demonstrated that solid sampling ETV-ICPMS is very well suited for the determination of metal traces in polyethylene. A limited multielement capability is often cited as an important drawback of ETV-ICPMS. However, by studying the effect of monitoring an increasing number of mass-to charge ratios on the signal profile (integrated signal intensity and repeatability) of selected analyte elements, the multielement capability of (solid sampling) ETV-ICPMS was systematically evaluated, and the results obtained suggest that, with a quadrupole-based ICPMS instrument, at least 11 elements can be determined "simultaneously" (from the same vaporization step), in essence without compromising the sensitivity or the precision of the results obtained. In this work, the "simultaneous" determination of Al, Ba, Cd, Cu, Mn, Pb, and Ti in a polyethylene candidate reference material has been accomplished, despite the large variation in analyte concentration (from 5 ng/g for Mn to 500 microg/g for Ti) and in furnace behavior (volatility) they exhibit. To avoid premature losses of Cd during thermal pretreatment of the samples, Pd was used as a chemical modifier. Two different calibration methods--external calibration using an aqueous standard solution and single standard addition--were studied and the results obtained were compared with those obtained using neutron activation analysis (NAA) and/or with the corresponding (candidate) certified values (if available). Single standard addition was shown to be preferable (average deviation between ICPMS result and reference value < 3%), although--except for Ba -acceptable results could also be obtained with external calibration. PMID- 11008766 TI - Monitoring electroosmotic flow by periodic photobleaching of a dilute, neutral fluorophore AB - Electroosmotic flow has been monitored in a capillary using a method based on periodic photobleaching of a neutral, fluorescent buffer additive. Rhodamine B was determined to be neutral between pH 6.0 and 10.8 and was added to the running buffer at a concentration of 400 nM. Rhodamine B was photobleached by opening a shutter under computer control for 250 ms every 5.00 s, to expose the dye to a laser beam and create a photobleached zone. The time was measured for the photobleached zone to migrate 6.13 mm to a downstream laser-induced fluorescence detector, to determine the rate of electroosmotic flow in the entire capillary. The flow rate was sampled every 5.00 s, and the precision of the flow measurements was 0.7% or better. Three fluorescent compounds were separated and detected by capillary electrophoresis with laser-induced fluorescence detection, while simultaneously monitoring the electroosmotic flow rate. PMID- 11008767 TI - Analysis of alkylphenol ethoxylate metabolites in the aquatic environment using liquid chromatography-electrospray mass spectrometry. AB - A quantitative method is described for the analysis of the metabolites of alkylphenol ethoxylate (APEO) surfactants in estuarine water and sediment samples using reversed-phase high-performance liquid chromatography with electrospray mass spectrometry detection. Nonyl- and octylphenols, nonyl- and octylphenol mono , di-, and triethoxylates, halogenated nonylphenols, and nonylphenol ethoxycarboxylates were concentrated from water samples using a C18 solid-phase extraction procedure. A novel, continuous-flow, high-temperature, sonicated extraction system was developed to isolate APEO metabolites from sediment samples. Quantitative LC-MS was performed in the negative ion mode for nonylphenols, octylphenols, and halogenated nonylphenols and in the positive ion mode for nonyl- and octylphenol ethoxylates using selected ion monitoring with isotopically labeled surrogate standards. Recoveries for sediment and water analyses ranged between 78 and 94%, and detection limits for APEO metabolites were between 1 and 20 pg injected on column. This is a significant improvement over previously reported methods. Suppression of analyte response was encountered in the presence of matrix components in sediment samples, but this effect was eliminated by careful selection of surrogate and internal standards. Individual APEO metabolite concentrations of 1-320 ng/L and 5-2000 ng/g are reported for water and sediment samples, respectively, from Jamaica Bay, NY. PMID- 11008768 TI - Modeling of sample dynamics in rectangular asymmetrical flow field-flow fractionation channels AB - We model the evolution of the concentration field of macromolecules in a rectangular asymmetrical flow field-flow fractionation channel using center manifold techniques. The deviation of the primary flow from a parabolic profile influences the concentration field and this is investigated to complement previously known results. The long-term evolution of the components of the sample is shown to be well described by a one-dimensional advection-diffusion equation. The coefficients of this equation are determined by the rigorous analysis of the complete set of equations governing the two-dimensional fluid flow. This model gives quantitative predictions of the elution time of the samples, the width of the concentration pulse, and the resolution of the apparatus. The influence of initial sample width and effects of the secondary relaxation from focusing to elution conditions are discussed. Reported theoretical predictions are in agreement with experimental results published previously. PMID- 11008769 TI - Comprehensive two-dimensional gas chromatography via differential flow modulation AB - A novel comprehensive two-dimensional gas chromatograph has been developed that utilizes differential flow modulation. This technique uses a 6-port valve to collect effluent from a primary column and periodically inject the effluent into a secondary column. The flow in the secondary column is kept 20 times larger than the flow in the primary column so the contents of the sample loop can be flushed into the secondary column in 5% of the collection time. Peaks widths at half maximum of approximately 0.06 s are generated for a 1.0 Hz secondary injection frequency. Sensitivity is not compromised, as 80% of the sample passes through both columns and reaches the detector. This simple yet effective technique has been used to analyze mixtures of alkanes, alkenes, aldehydes, alcohols, aromatics, esters, and ketones with high speed and high resolution. PMID- 11008770 TI - Monte Carlo model of nonlinear chromatography AB - A stochastic approach to the nonlinear chromatography theory, based on the Monte Carlo simulation method, is presented. A computer program, acting as a "virtual chromatograph" and performing a discrete event simulation, is described. Such a program allows one to choose the column type, operating conditions, sample composition, injection method, mobile-phase dispersion model, and stationary phase sorption-desorption kinetics. Nonlinearity is accounted for by continuously monitoring and updating both the column and the solute status and by moving individual molecules step by step along the column according to specific random modes. The program has been validated through a series of statistical tests and comparing the results with the well-known achievements of the classical stochastic theory. A first application is presented, referred to a real case benzene elution on a gas solid capillary column, where the Langmuir adsorption isotherm is assumed. The effect of both the sorption modes and the site capacity are investigated. Possible applications to investigate open problems in several fields of separation science are emphasized. In addition, several specific points such as the down-scaling of a real case and the correspondence of specific adsorption dynamics with the equilibrium Langmuir isotherm are described. PMID- 11008771 TI - Resolving and quantifying overlapped chromatographic bands by transmutation AB - A new chemometric technique called "transmutation" is developed for the purpose of sharpening overlapped chromatographic bands in order to quantify the components. The "transmutation function" is created from the chromatogram of the pure component of interest, obtained from the same instrument, operating under the same experimental conditions used to record the unresolved chromatogram of the sample mixture. The method is used to quantify mixtures containing toluene, ethylbenzene, m-xylene, naphthalene, and biphenyl from unresolved chromatograms previously reported. The results are compared to those obtained using window factor analysis, rank annihilation factor analysis, and matrix regression analysis. Unlike the latter methods, the transmutation method is not restricted to two-dimensional arrays of data, such as those obtained from HPLC/DAD, but is also applicable to chromatograms obtained from single detector experiments. Limitations of the method are discussed. PMID- 11008772 TI - Multidimensional analysis of poly(ethylene glycols) by size exclusion chromatography and dynamic surface tension detection AB - Substantial improvements in a multidimensional dynamic surface tension detector (DSTD) are presented. Rapid, online calibration and measurement of the dynamic surface tension for high-performance liquid chromatography separations is achieved. Dynamic surface tension is determined by measuring the differential pressure across the liquid-air interface of repeatedly growing and detaching drops. Continuous surface tension measurement throughout the entire drop growth (50 ms to 2 s) is achieved, for each eluting drop, providing insight into the kinetic behavior of molecular orientation processes at the liquid-air interface. Three-dimensional data are obtained, with surface tension first converted to surface pressure, which is plotted as a function of elution time axis versus drop time axis. Two key innovations will be reported. First, a novel calibration procedure is described and implemented. Differential pressure signals from three drops (mobile phase, standard in mobile phase, and analyte in mobile phase) are utilized to make the dynamic surface tension measurement, thereby eliminating the need for optical imaging, and viscosity and hydrostatic pressure corrections, as required by other methods. Only pressure signals from one mobile-phase drop and one standard drop pressure signal are required, while the analyte drop pressure signal is measured along the chromatographic time axis. Second, corrections for drop elongation are not required, because the drops are precisely detached by an air burst actuation method in a regime were the surface tension forces significantly dominate gravitational forces. Drops that would fall with a volume of approximately 10 microL due to gravity are precisely and repeatedly detached earlier at a volume of 2 microL. The sensitivity and unique selectivity of the DSTD opens up new possibilities in the analysis of small molecular weight polymers of varying degrees of surface activity, as illustrated for the size exclusion chromatography analyses of complex poly(ethylene glycol) (PEG) samples. Using partial least squares for data analysis, polydispersity of complex PEG samples is determined at a relative precision of approximately 1%. PMID- 11008773 TI - Chemical grafting of molecularly imprinted homopolymers to the surface of microplates. Application of artificial adrenergic receptor in enzyme-linked assay for beta-agonists determination. AB - A technique for coating of microplate wells with a molecularly imprinted polymer (MIP), specific for epinephrine, is presented. 3-Aminophenylboronic acid was polymerized in the presence of epinephrine using oxidation of the monomer by ammonium persulfate. This process resulted in the grafting of a thin polymer layer onto the polystyrene surface of the microplates. The polymer affinity was determined by an enzyme-linked assay using a conjugate of horseradish peroxidase and norepinephrine (HRP-N). It was found that imprinting resulted in increased affinity of the polymer toward HRP-N and epinephrine. Influence of the buffer pH and concentration on the polymer affinity was analyzed. It was shown that the MIP coated microplates could be used for assay development and drug screening. The high stability of the polymers and good reproducibility of the measurements make MIP coating an attractive alternative to traditional antibodies or receptors, used in ELISA. PMID- 11008774 TI - High-performance liquid chromatography-electrospray ionization mass spectrometry of single- and double-stranded nucleic acids using monolithic capillary columns. AB - Monolithic capillary columns were prepared by copolymerization of styrene and divinylbenzene inside a 200-microm i.d. fused silica capillary using a mixture of tetrahydrofuran and decanol as porogen. With gradients of acetonitrile in 100 mM triethylammonium acetate, the synthesized columns allowed the rapid and highly efficient separation of single-stranded oligodeoxynucleotides and double-stranded DNA fragments by ion-pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC). Compared with capillary columns packed with micropellicular, octadecylated poly-(styrene/divinylbenzene) particles, an improvement in column performance of approximately 40% was obtained, enabling the analysis of an 18-mer oligodeoxynucleotide with a column efficiency of more than 190000 plates per meter. The chromatographic separation system was on-line-coupled to electrospray ionization mass spectrometry (ESI-MS). To improve the mass spectrometric detectabilities, 25 mM triethylammonium bicarbonate was utilized as an ion-pair reagent at the cost of only little reduction in separation performance and acetonitrile was added postcolumn as the sheath liquid through the triaxial electrospray probe. High-quality mass spectra of femtomole amounts of 3-mer to 80 mer oligodeoxynucleotides were recorded showing very little cation adduction. Double-stranded DNA fragments ranging in size from 51 to 587 base pairs were separated and detected by IP-RP-HPLC-ESI-MS. Accurate mass determination by deconvolution of the mass spectra was feasible for DNA fragments up to the 267 mer with a molecular mass of 165 019, whereas the spectra of longer fragments were too complex for deconvolution because of incomplete separation due to overloading of the column. Finally, on-line IP-RP-HPLC tandem MS was applied to the sequencing of short oligodeoxynucleotides. PMID- 11008775 TI - Evaluation of a vancomycin chiral stationary phase in capillary electrochromatography using polar organic and reversed-phase modes. AB - A vancomycin chiral stationary phase (CSP) was fully evaluated in capillary electrochromatography (CEC) in reversed-phase and polar organic modes for a number of racemic pharmaceutical compounds. High efficiency and resolution values were obtained for a number of compound classes including thalidomide in both the polar organic mode (190000 plates meter(-1) and Rs = 13.8) and reversed-phase mode (125000 plates meter(-1) and Rs = 13.0). Experimental parameters, including organic modifier, organic solvent ratio, ionic strength, pH, temperature, and voltage, were examined in both the aqueous and nonaqueous modes to deduce their effect on the resultant EOF, retention times, resolution, and efficiency of chiral separations. All results were consistent with and found to be a combination of what is known from existing literature on CEC theory and experience obtained with macrocyclic antibiotic CSPs in LC. Column stability was excellent, and each column packed was found to offer repeatable separations even when switching from the aqueous to the nonaqueous mode. PMID- 11008776 TI - Determination of arsenic compounds by high-performance liquid chromatography ultrasonic nebulizer-high power nitrogen-microwave-induced plasma mass spectrometry: an accepted coupling. AB - To establish a sensitive, accurate, and precise determination of arsenic compounds, a high power nitrogen microwave-induced plasma (1.3 kW) mass spectrometer (N2-MIP-MS) has been successfully coupled with an ultrasonic nebulizer (HPLC-USN) that is attached to a high-performance liquid chromatograph. It is examined as an element-specific detector for its applicability to the optimization and determination of seven arsenic compounds [arsenic acid, methylarsonic acid (MA), dimethylarsinic acid (DMA), arsenobetaine (AB), arsenocholine (AC), trimethylarsine oxide (TMAO), and tetramethylarsonium ion (CMI)]. This HPLC-USN-MIP-MS coupling is an encouraging combination as an alternative method for mass spectroscopy for elemental speciation analysis. Interchanging of the MIP-MS fabricated nebulizer (concentric) with an ultrasonic nebulizer, increases 3-6 times the ion signals for the anionic and 6-12 times those for the cationic arsenic compounds as compared to traditional methods. The HPLC-USN-MIP-MS combination used is excellent, amplifying the ion signals about 1.5-2 times for cationic and 1.3-2.8 times for the anionic arsenic compounds as compared to the HPLC-ICPMS coupling. The detection limits for As(V), MA, DMA, AB, TMAO, AC, and TMI (in Milli-Q-water) obtained with the optimized HPLC-USN-N2-MIP MS system are 0.46, 0.36, 0.73, 0.21, 3.64, 0.39, and 0.32 microg L(-1), respectively, about 13-50 times lower than the HPLC-MIP-MS and about 3-11 times lower than the HPLC-ICPMS. The detection limits of As(V), MA, DMA, AB, TMAO, AC, and TMI, which spike in the urine, are deteriorated by 1.7-4.2 times compared with the detection limits of the seven different As compounds, which are prepared in the Milli-Q-water. The repeatability (RSD for three successive analyses) and reproducibility (RSD for three successive analyses performed on three different days), considering peak area and peak height, achieved for seven different arsenic compounds are 0.5-7 and 0.7-8%, comparable with the HPLC-ICPMS (0.3-8.5%; 4-12%) and HPLC-MIP-MS (0.4-9%; 5-12%) systems. The combined HPLC-USN-N2-MIP-MS has been adequately applied to the determination of AB in NIES Candidate Human Urine CRM. The results agree with the HPLC-ICPMS values. Chloride interference as 40Ar35Cl+ is not found in the urine and with the high chloride matrix (10000 mg L(-1)). PMID- 11008777 TI - Quaternized trimethylaminated polystyrene-coated zirconia as a strong anion exchange material for HPLC. AB - The synthesis and characterization of a new, base-stable, strong anion exchange phase by amination of polystyrene-coated zirconia (PS-ZrO2) are described. Even though the ion exchange capacity of the quaternized trimethylaminated PSZrO2 (QTMA-PS-ZrO2) is only 0.07 mequiv/g, it is able to separate various inorganic anions, benzoic acid derivatives, and nucleotides in their deprotonated states. The effects of ionic strength, eluent pH, and counterion type are discussed. In the presence of both phosphate and fluoride ions in the eluent, band broadening caused by Lewis acid/base interactions between zirconia and analytes is greatly suppressed. The mixed retention modes (ion exchange, hydrophobic interaction, and Lewis acid/base interactions) on QTMA-PS-ZrO2 offer a different selectivity toward various anionic analytes than do other zirconia- and nonzirconia-based ion exchangers. PMID- 11008778 TI - Functionalization of a poly(amidoamine) dendrimer with ferrocenyls and its application to the construction of a reagentless enzyme electrode. AB - Poly(amidoamine) dendrimers having various degrees of modification with the redox active ferrocenyls were prepared by controlling the molar ratio of ferrocenecarboxaldehyde to amine groups of dendrimers. By alternate layer-by layer depositions of partial ferrocenyl-tethered dendrimers (Fc-D) with periodate oxidized glucose oxidase (GOx) on a Au surface, an electrochemically and enzymatically active multilayered assembly of enzyme was constructed. The resulting GOx/Fc-D multilayer-associated electrodes were electrochemically analyzed, and the surface concentration of ferrocenyl groups, active enzyme coverage, and sensitivity were estimated. A 32% dendrimer modification level of surface amines to ferrocenyls was found to be an optimum in terms of enzyme dendrimer network formation, electrochemical interconnectivity of ferrocenyls, and electrode sensitivity. With the prepared Fc(32%)-tethered dendrimers, mono- and multilayered GOx/Fc-D electrodes were constructed, and their electrochemical and catalytic properties were characterized. The bioelectrocatalytic signals from the multilayered GOx/Fc-D electrodes were shown to be directly correlated to the number of deposited bilayers. From this result, it seems that the electrode sensitivity is directly controllable, and the multilayer-forming strategy with partial ferrocenyl-tethered dendrimers is useful for the construction of reagentless biosensors. PMID- 11008779 TI - Nanoparticle characterization in nanoliter volumes by grating light reflection spectroscopy AB - We present both theoretical and experimental results demonstrating that grating light reflection spectroscopy (GLRS) can provide information about the concentration and average size of particles of nanometer dimensions distributed in liquid-phase media. To demonstrate this, we have performed experiments on various concentrations of dendrimeric oligomers in water. Our results show that, with GLRS, we can determine the mean radius of particles with sizes on the order of molecular dimensions. The measurements were carried out in a continuous-flow format using a microchannel flow system and in a detection volume of less than 200 nL PMID- 11008780 TI - Reduction of mass bias and matrix effects in inductively coupled plasma mass spectrometry with a supplemental electron source in a negative extraction lens AB - Electrons from a heated tungsten filament are created inside the extraction lens and driven out toward the skimmer. These electrons move through the ion path and reduce space charge effects between positive ions in the beam. The ion transmission efficiency is improved by factors of two (for Pb+) to 27 (for Li+). The greater sensitivity improvement for low-mass ions leads to a substantial reduction in mass bias. With the additional electrons, MO+/M+ and M2+/M+ abundance ratios increase but can be minimized with a small reduction in aerosol gas flow rate. No new background ions are observed with this technique. Matrix effects can be significantly diminished when the electron source is operated under the high electron current mode. The mass dependence of matrix-induced suppression of analyte signals is essentially eliminated. Using flow injection analysis to minimize solid deposition, the technique can tolerate Na matrix up to 10000 ppm (1%) with only approximately 15% loss of analyte sensitivity. PMID- 11008781 TI - Electrochemical characterization of electrodes with submicrometer dimensions AB - The construction and electrochemical characterization of electrodes with submicrometer dimensions (2 nm < r(app) < 1000 nm) is reported. Electrodes are prepared by insulating etched Pt wires with electrophoretic paint, as recently reported by Slevin et al. (Electrochem. Commun. 1999, 1, 282). The voltammetric behavior of these electrodes was evaluated using nine different redox systems; well-defined and stable diffusion-limited responses were obtained in all but two cases. The behavior of these electrodes was investigated in aqueous ferrocenylmethyltrimethylammonium (FcTMA+) solutions in the presence and absence of excess supporting electrolyte to determine the influence of diffusion and migration on molecular transport in the nanometer spatial regime. Our findings indicate that the voltammetric behavior of these electrodes can be described using classical transport theory for r(app) > 10 nm. PMID- 11008782 TI - Direct in situ measurements of labile inorganic and organically bound metal species in synthetic solutions and natural waters using diffusive gradients in thin films. AB - The emerging technique of DGT (diffusive gradients in thin films) is shown to be capable of performing new speciation measurements in situ in natural waters. In DGT, metals are bound to a resin layer after passing through a well-defined diffusion layer. Cd was measured in solutions containing glycine, EDTA, and fulvic (FA) and humic acids (HA) by atomic absorption spectroscopy (AAS), anodic stripping voltammetry (ASV), and DGT. DGT measured similar labile fractions to ASV, with detailed differences being consistent with a thicker diffusion layer allowing more dissociation of labile complexes and a slower diffusion of FA and HA complexes through the gel. When single measurements are made in complex solutions with DGT, precise quantification is impossible due to uncertainties concerning the distribution of species with different diffusion coefficients. A new procedure was proposed based on the advantage of DGT of being able to control the pore size of the diffusive gel layer. Small (inorganic) species diffuse freely through all gels but larger FA and HA (organic) complexes diffuse less freely in more constrained gels. When measurements were made on known solutions of Cu and FA or HA, it was possible to quantify the inorganic and organic species separately. They agreed well with predictions made using the WHAM speciation code. Multiple DGT units were also deployed in situ in a stream with high dissolved organic carbon (14.6 mg/L). The systematic differences between the devices with different gel compositions enabled determination, for the first time, of the in situ concentrations of both labile inorganic and organic species in natural water. A single DGT device with a constrained gel can be used to quantify inorganic species directly, providing absolute accuracy is not required. This ability of DGT to measure well-defined fractions of metals in situ using a simple device gives it considerable potential as a regulatory tool. A direct speciation measurement may be preferable to modeling approaches which require diverse input data that are difficult to determine. PMID- 11008783 TI - Determination of hydroxytyrosol in plasma by HPLC. AB - Hydroxytyrosol (2-(3,4-dihydroxyphenyl)ethanol), a phenolic compound present in extravirgin olive oil, has been reported to contribute to the prevention of cardiovascular disease. The present study describes an accurate and reproducible reversed-phase HPLC method to measure hydroxytyrosol in plasma. This compound was extracted from acidified plasma by solid-phase extraction using an Oasis HLB copolymer. The plasma sample was rinsed with water and methanol in water (5:95; v/v). Hydroxytyrosol was eluted with methanol, which was subsequently evaporated under a nitrogen stream. Analysis by HPLC with diode array-UV detection was carried out using a C18 column and a gradient elution with acidified water and methanol/acetonitrile (50:50; v/v). The method was validated by the analyses of plasma samples spiked with pure hydroxytyrosol, obtaining a linear correlation (0.9986) and precision with a coefficient of variation ranging from 0.79 to 6.66%. The recovery was approximately 100%, and the limit of detection was 37 ng/mL. The oral administration of hydroxytyrosol to rats and its subsequent detection in plasma showed that the method is suitable for pharmacokinetic studies. PMID- 11008784 TI - Continuous-flow microextraction exceeding 1000-fold concentration of dilute analytes AB - A novel liquid-liquid microextraction method, that we have termed continuous-flow microextraction (CFME), is described. In a 0.5-mL glass chamber, an organic drop (1-5 microL) is held at the outlet tip of a polyetheretherketone (PEEK) connecting tubing which is immersed in a continuously flowing sample solution and acts as the fluid delivery duct and as a solvent holder. Extraction takes place between the organic drop and the flowing sample solution that is continuously ejected out of the PEEK tubing. Concentration factors of between 260- to 1600 fold are achieved within 10 min of extraction. Aspects relevant to CFME were studied. In combination with gas chromatography-electron capture detection, CFME allows analytes to be detected at femtogram-per-milliliter levels. The performance of this technique was evaluated on the basis of the analysis of trace nitroaromatic compounds and chlorobenzenes in environmental samples. PMID- 11008785 TI - A planar pCO2 sensor with enhanced electrochemical properties. AB - To develop planar microchemical pCO2 sensing devices with improved electrochemical properties, we combined two advanced technologies. One is a differential sensor arrangement to simplify the microfabrication procedure by employing pH-sensitive gas-permeable membranes, and the other is the use of an enzyme (carbonic anhydrase) to shorten total measurement time by accelerating the rate of CO2 hydration. The adhesion of the polyurethane-matrix gas-permeable membrane is enhanced significantly by incorporating a silanizing reagent (silicon tetrachloride), improving the stability and extending sensor lifetime. The proposed differential pCO2 microelectrodes exhibited significantly improved performance in their preconditioning period, response and recovery times, stability, response slope, and lifetime. PMID- 11008786 TI - Rapid identification of the bacterial pathogens responsible for urinary tract infections using direct injection CE. AB - The use of high-performance capillary electrokinetic techniques for the separation, identification, and quantitation of intact microbes represents a new frontier for separation science. In this work, it is demonstrated that pathogens most responsible for urinary tract infections can be distinguished from one another after direct injection of untreated urine. High efficiencies (often exceeding 1000000 plates/m) and short analysis times (< 10 min) are characteristics of this approach. The concentration of the urine matrix appears to be able to cause a small, but definite, change in the electroosmotic flow velocity. This high-efficiency separation-based approach could prove to be invaluable for the diagnosis and tracking of certain diseases. It also could form the basis for a variety of rapid microbial assays. PMID- 11008787 TI - Environmental mass spectrometry. PMID- 11008788 TI - Dynamic electrochemistry: methodology and application. PMID- 11008790 TI - Kenneth Stevens receives National Medal of Science PMID- 11008792 TI - Reviews of acoustical patents PMID- 11008789 TI - Chiral separations. PMID- 11008791 TI - ASA presents special awards to young scientists at the 51st annual International Science and Engineering Fair (ISEF). Acoustical Society of America PMID- 11008793 TI - A three-dimensional, two-way, parabolic equation model for acoustic backscattering in a cylindrical coordinate system AB - A new PE model for solving three-dimensional, forward and backward sound propagation in a cylindrical coordinate system is presented. The model marches a wave field in the radial direction including the azimuthal diffraction effects, and solves for a backscattered field based on a three-dimensional, single scattering approach. A periodic sidewall boundary condition is applied for computations in a 360-degree sector, while an approximate sidewall boundary condition is used for calculation in a sector less than 360 degrees. These two sidewall boundary conditions are verified by the numerical results. The major drawback of using the cylindrical coordinate system, when the backscattering solution is valid within a limited area, is analyzed using a geometrical-optical interpretation. The model may be useful for studying three-dimensional backscattering phenomena comprising azimuthal diffraction effects. PMID- 11008794 TI - Quantitative imaging using a time-domain eigenfunction method AB - An inverse scattering method that uses eigenfunctions of a scattering operator at a single frequency is extended to include the full range of frequencies present in the incident pulse waveform. The resulting so-called time-domain eigenfunction method is shown to yield a modulated version of the scattering potential. The potential is recovered by a demodulation process using cross correlation with a reference. Including an adaptive delay in the reference is shown to compensate partially for the linearization of the Born approximation and to extend its valid range. The k-space window of the time-domain solution is expressed in terms of the incident waveform and shown to be smoother than that of a single-frequency solution. The time-domain method is examined using both calculated and measured data. In the calculations, an exact solution for scattering from one or multiple nonconcentric cylinders is used to obtain the scattered field. In the measurements, a novel ring-transducer system was employed to obtain the incident and total fields. The results of simulations and experiments show that the method is robust and accurate for the size of objects considered and that the point resolution approaches one-half the wavelength at the pulse center frequency. PMID- 11008795 TI - Scattering of antiplane shear waves by a circular cylinder in a traction-free plate AB - Following a well-established formula used by many researchers, the scattering of an anti-plane shear wave by an infinite elastic cylinder of arbitrary relative radius centered in a traction-free two-dimensional isotropic plate has been examined. The plate is divided into three regions by introducing two imaginary planes located symmetrically away from the surface of the cylinder and perpendicular to surfaces of the plate. The wave field is expanded into cylinder wave modes in the central bounded region containing the cylinder, while the fields in the other two outer regions are expanded into plate wave modes. A system of equations determining the expansion coefficients is obtained according to the traction-free boundary conditions on the plate walls and the stress and displacement continuity conditions across the imaginary planes. By taking an appropriate finite number of terms of the infinite expansion series and a few selected points on the two properly chosen virtual planes and the surfaces of the plate through convergence and precision tests, a matrix equation to numerically evaluate the expansion coefficients is found. The method of how to choose the locations of the imaginary planes and the terms of the expansion series as well as the points on each respective boundary is given in Sec. III in detail. Curves of the reflection and transmission coefficients against the relative radius of the cylinder in welded and slip or cracked interfacial conditions are shown. Analysis on the contrast variations of the reflection and transmission coefficients for a cylinder in bonded and debonded interfacial situations is made. The relative errors estimated by the deviation of the numerical results from the principle of the conservation of energy are found to be less than 2%. PMID- 11008796 TI - On approximating guided waves in plates with thin anisotropic coatings by means of effective boundary conditions AB - In this paper, effective boundary conditions for elastic wave propagation in plates with thin coatings are derived. These effective boundary conditions are used to obtain an approximate dispersion relation for guided waves in an isotropic plate with thin anisotropic coating layers. The accuracy of the effective boundary conditions is investigated numerically by comparison with exact solutions for two different material systems. The systems considered consist of a metallic core with thin superconducting coatings. It is shown that for wavelengths long compared to the coating thickness there is excellent agreement between the approximate and exact solutions for both systems. Furthermore, numerical results presented might be used to characterize coating properties by ultrasonic techniques. PMID- 11008797 TI - Acoustic streaming in annular thermoacoustic prime-movers AB - The theory of acoustic streaming in an annular thermoacoustic prime-mover is developed. It is predicted that above the threshold for traveling wave excitation the device considered (which does not contain any moving parts or externally imposed pressure gradients) produces circulation of fluid. The heat flux carried by this directional mass flow inside the thermoacoustic stack exceeds (or is comparable with) the heat flux associated with the acoustically induced increase of thermal diffusivity of the gas. The effects investigated are important for optimization of the performance of thermoacoustic devices. PMID- 11008798 TI - Measurement of a third-order elastic constant in silicon by a comparison method AB - The third-order elastic constant C111 of silicon is measured with a comparison method, based on the interaction between longitudinal acoustic waves. The value is in good agreement with published values and demonstrates the validity of the method. PMID- 11008799 TI - Effective impedance of rough porous ground surfaces AB - Laboratory measurements at grazing angles of the impedance of sand surfaces containing artificial two-dimensional (2D) roughness are compared with predictions of an extended boss theory, which allows for incoherent scatter, randomness, and finite impedance effects. The data and predictions are found to be in reasonable agreement. The rough surface impedance model may be used, as an alternative to layering, to explain measurements where the reactance exceeds the resistance at low frequencies. The model also explains instances where the measured ground impedance tends to zero at higher frequencies. In addition, the model is found to predict features of measurements made with an elevated source and vertically separated receivers at a range of 30 m over cultivated ground surfaces. PMID- 11008800 TI - Acoustic field variability induced by time evolving internal wave fields AB - A space- and time-dependent internal wave model was developed for a shallow water area on the New Jersey continental shelf and combined with a propagation algorithm to perform numerical simulations of acoustic field variability. This data-constrained environmental model links the oceanographic field, dominated by internal waves, to the random sound speed distribution that drives acoustic field fluctuations in this region. Working with a suite of environmental measurements along a 42-km track, a parameter set was developed that characterized the influence of the internal wave field on sound speed perturbations in the water column. The acoustic propagation environment was reconstructed from this set in conjunction with bottom parameters extracted by use of acoustic inversion techniques. The resulting space- and time-varying sound speed field was synthesized from an internal wave field composed of both a spatially diffuse (linear) contribution and a spatially localized (nonlinear) component, the latter consisting of solitary waves propagating with the internal tide. Acoustic simulation results at 224 and 400 Hz were obtained from a solution to an elastic parabolic equation and are presented as examples of propagation through this evolving environment. Modal decomposition of the acoustic field received at a vertical line array was used to clarify the effects of both internal wave contributions to the complex structure of the received signals. PMID- 11008801 TI - Tomographic inversion for sediment parameters in shallow water AB - This article discusses inversions for bottom geoacoustic properties using broadband acoustic signals obtained from explosive sources. The experimental data used for the inversions are SUS charge explosions acquired on a vertical hydrophone array during the Shelf Break Primer Experiment conducted south of New England in the Middle Atlantic Bight in August 1996. The SUS signals were analyzed for their time-frequency behavior using wavelets. The group speed dispersion curves were obtained from the wavelet scalogram of the SUS signals. A genetic algorithm (GA) was used for the inversion of sound speeds in the water column and compressional wave speeds in the sediment layers. The variations in the sound speeds in the water column were represented using empirical orthogonal functions (EOFs). A range-independent normal mode routine was used to construct the replica fields corresponding to the parameters. Comparison of group speeds for modes 1 to 9 and for a range of frequencies 8 to 200 Hz was used to arrive at the best parameter fit. An efficient hybrid optimization scheme using the GA and a Levenberg-Marquardt algorithm is presented. Linear perturbation methods were also used to "fine tune" the inversions and to obtain resolution and variance estimates. Analysis was also done to compute the degree of convergence of each of the parameters by explicitly calculating the Hessian matrices numerically. A posteriori estimation of mean and covariance was also done to obtain error estimates. Group speeds for the inverted sound speed fields provide an excellent match to the experimental data. The inverted sediment compressional speed profile compares well with in situ measurements. PMID- 11008802 TI - In situ estimation of sediment sound speed and critical angle AB - Understanding the basic physics of sound penetration into ocean sediments is essential for the design of sonar systems that can detect, localize, classify, and identify buried objects. In this regard the sound speed of the sediment is a crucial parameter as the ratio of sound speed at the water-sediment interface determines the critical angle. Sediment sound speed is typically measured from core samples using high frequency (100's of kHz) pulsed travel time measurements. Earlier experimental work on subcritical penetration into sandy sediments has suggested that the effective sound speed in the 2-20 kHz range is significantly lower than the core measurement results. Simulations using Biot theory for propagation in porous media confirmed that sandy sediments may be highly dispersive in the range 1-100 kHz for the type of sand in which the experiments were performed. Here it is shown that a direct and robust estimate of the critical angle, and therefore the sediment sound speed, at the lower frequencies can be achieved by analyzing the grazing angle dependence of the phase delays observed on a buried array. A parametric source with secondary frequencies in the 2-16 kHz range was directed toward a sandy bottom similar to the one investigated in the earlier study. An array of 14 hydrophones was used to measure penetrated field. The critical angle was estimated by analyzing the variations of signal arrival times versus frequency, burial depth, and grazing angle. Matching the results with classical transmission theory yielded a sound speed estimate in the sand of 1626 m/s in the frequency range 2-5 kHz, again significantly lower the 1720 m/s estimated from the cores at 200 kHz. However, as described here, this dispersion is consistent with the predictions of the Biot theory for this type of sand. PMID- 11008803 TI - Measurement technique for bottom scattering in shallow water AB - Sonar performance predictions of reverberation in shallow water rely upon good estimates of the bottom-scattering strength. However, little is understood about bottom scattering in shallow water in the frequency range 400-4000 Hz, particularly its dependency upon frequency and its relationship to the physical properties of the seafloor. In order to address these issues, a new measurement technique has been developed to probe the frequency and angular dependency of bottom-scattering strength. The experimental technique is described which employs either coherent or incoherent sources (lightbulbs). In addition, measurement and modeling results for two diverse shallow water sites are presented. At one site, the scattering appears to arise at or near the water-sediment interface. At the other site, scattering from a 23-m sub-bottom horizon is clearly apparent in the data at and below 1800 Hz. The fact that our measurement technique can directly reveal the presence of sub-bottom scattering is a significant advance in the development of methods to explore the physical mechanisms that control bottom scattering. PMID- 11008804 TI - Low-grazing-angle monostatic acoustic reverberation from rough and heterogeneous seafloors AB - Numerical wavefield modeling, based on time domain finite difference solutions to the full elastic wave equation, is used to quantitatively predict the sensitivity of monostatic backscatter to variations in geological properties of the seabed. This article addresses the hypothesis that observed backscatter signals from the seafloor are produced by a combination of seafloor (interface) and subseafloor (volume) scattering from structures having variations at scale lengths comparable to the wavelength of the insonifying acoustic field. Wavelength-scale seafloor roughness and subseafloor volume heterogeneity parameters are defined using stochastic spatial probability functions having Gaussian autocorrelations. The range of the variations in these parameters is constrained to realistic values based on estimates derived from seafloor bathymetry and other geologic data. Modeling results show that backscattering from rough, basaltic (hard) bottoms, in the absence of large-scale seafloor slope, is dominated by rough surface scattering with little contribution from volume scattering. Contrary to this, sediment (soft) bottoms with subseafloor volume heterogeneity, with or without seafloor roughness, produce significant backscattering signals compared to a homogeneous sediment bottom. PMID- 11008805 TI - Resonant response of complex shell structures AB - Resonances of shell structures play a prominent role in determining their acoustic characteristics. Among the most important of such characteristics are radiated noise, scattering, and self-noise levels. The nature of the structural resonances, for example the number of modes within a particular frequency band or the overall spatial dependence of the response, determines the importance of these phenomena in various contexts and what steps may be taken to modify their acoustic impact. In the past, knowledge of the resonant response of highly idealized systems has been used to guide the development of quiet platforms. The results of a scale model investigation of the effects of increased complexity on the nature of the resonances of submerged shell structures are reported. Effects presented here are flexural Bloch mode resonances, localized resonances resulting from structural irregularity, and the effects of non-axisymmetry on these phenomena. The implications of these results on acoustic design and the development of systems is discussed. PMID- 11008806 TI - Sensor scheme design for active structural acoustic control AB - Efficient sensing schemes for the active reduction of sound radiation from plates are presented based on error signals derived from spatially weighted plate velocity or near-field pressure. The schemes result in near-optimal reductions as compared to weighting procedures derived from eigenvector or singular vector analysis of the radiation operator. Efficient control configurations are suggested using a, possibly analog, front-end implementing a bank of spatial weighting functions and a digital controller with a minimized number of input and output channels. The performance of different weighting functions is compared, as well as the performance of different frequency-dependent filtering functions. Design rules are given for the sensor spacing, the number of weighting functions, the number of actuators, and the corresponding controller dimensionality. PMID- 11008807 TI - The vibration behavior of railway track at high frequencies under multiple preloads and wheel interactions AB - The track foundation is preloaded by multiple wheel loads due to the train weight and, as the pad and ballast are nonlinear, their stiffness depends upon the preload in them. Due to the influence of these resilient components of the track, the track vibration is affected by the wheel loads. It is also affected by the wheel/rail interactions. In this article the preloads in the pad and ballast are calculated by considering the nonlinear properties of the track foundation, and thus the preloaded pad and ballast stiffnesses are determined. The vibration properties are explored for the track under multiple wheel loads and multiple wheel/rail interactions by comparing the results from different track models with and without these effects. It is found that the point receptance of the track is reduced and the vibration decay rate is enhanced at low frequencies due to the wheel loads. The effects of the wheel/rail interactions are most significant for frequencies 400-2000 Hz. Because of the wheel/rail interactions, the point receptance fluctuates and the vibration decay is enhanced in the regions around the wheels. PMID- 11008808 TI - European methodology for testing the airborne sound insulation characteristics of noise barriers in situ: experimental verification and comparison with laboratory data AB - In the frame of the 1994-1997 Standard, Measurement and Testing program, the European Commission funded a research project, named Adrienne, to define new test methods for measuring the intrinsic characteristics of road traffic noise reducing devices in situ. The research team produced innovative methods for testing the sound reflection/absorption and the airborne sound insulation characteristics of noise barriers. These methods are now under consideration at CEN (European Committee for Standardization), to become European standards. The present work reports a detailed verification of the test method for airborne sound insulation over a selection of 17 noise barriers, representative of the Italian and European production. The samples were tested both outdoors, using the new Adrienne method, and in laboratory, following the European standard EN 1793 2. In both cases the single number rating for airborne sound insulation recommended by the European standard was calculated. The new method proved to be easy to use and reliable for all kinds of barriers. It has been found sensitive to quality of mounting, presence of seals, and other details typical of outdoor installations. The comparison between field and laboratory results shows a good correlation, while existing differences can be explained with the different sound fields and mounting conditions between the outdoor and laboratory tests. It is concluded that the Adrienne method is adequate for its intended use. PMID- 11008809 TI - The effect of high-amplitude sound on the attenuation of perforated tube silencers AB - A time-domain computational approach is applied to investigate the behavior of perforated tube silencers at high sound levels. The one-dimensional computational technique employs a lumped parameter model for the perforate flows. The lumped parameter perforate model is based on time-invariant approximations for the equivalent length l(eq) and resistance R, consistent with the use of a nonlinear perforate impedance. Empirical expressions for l(eq) and R are developed experimentally using nondimensional scaling parameters. The model is applied to geometries representative of automotive resonators and multiple-duct mufflers. Conditions are simplified from those in an actual automotive system by considering single-frequency excitation and zero mean flow. Simulations with linear perforate behavior are compared to experimental data obtained with an extended impedance tube system. Simulations with nonlinear perforate behavior for one concentric tube resonator are compared to published experimental data. PMID- 11008810 TI - Interference from degraded auditory stimuli: linear effects of changing-state in the irrelevant sequence. AB - Cognitive performance, particularly on a number of tasks involving short-term memory for order, is impaired by the mere presence of irrelevant background sound. The current study examines the features of the irrelevant sound that determine its disruptive potency. Previous research suggests that the amount of variability in an irrelevant stream is related to the degree of disruption of memory. The present experiments used a parametric approach to manipulate degree of change more precisely. Increasing levels of degradation, effected either by low-pass filtering (speech) or by digital manipulation (speech and nonspeech), monotonically decreased the degree of interference. The findings support the following propositions: (i) the degree of physical change within an auditory stream is the primary determinant of the degree of disruption; and, (ii) the effects of irrelevant speech and irrelevant nonspeech sounds are functionally similar. PMID- 11008811 TI - Acoustic optic hybrid (AOH) sensor AB - The ability of laser vibrometers to receive and process acoustic echoes from the water surface above a submerged target is established and evaluated. Sonar echoes from a submerged target are collected from the water surface by a laser vibrometer. Feasibility of this approach to sensing underwater sound is demonstrated. If the acoustic excitation at an otherwise undisturbed water surface is 195 to 168 dB re: 1 microPa, signal-to-noise ratio (SNR), at the vibrometer output, is shown to range from about 46 to 6 dB. Capillary waves and gravity waves at the water surface are expected and shown to have some destructive effect on the process of echo retrieval. A series of experiments to quantify the surface wave effects is described. The wave experiment results are reported. A successful attempt to acquire echoes from a submerged target over a grid of points for further processing into a three-dimensional image is made and described. The data acquisition and beamforming techniques constitute a three dimensional, acoustic optic, synthetic aperture sonar (SAS). Beamformed images are included. For an aircraft towing acoustic sensors through the water with a mechanical link, this technique holds the promise of increased safety and improved fuel efficiency. PMID- 11008812 TI - Acoustic impedances at the oval window, and sound pressure transformation of the middle ear in Norwegian cattle. AB - In 15 cadaver ears from Norwegian cattle, sound pressure transfer functions have been measured (1) for sound input to the tympanic membrane, (2) for sound input to the oval window with the footplate in place, but with the ossicular chain removed, and (3) for sound input to the oval window with also the footplate removed. The output pressure was measured in an enclosure cemented to the round window. The data allow calculation of equivalent sound pressures at the input positions, as well as the acoustic input impedances at the oval window with intact footplate, Z(sc), and with the footplate removed, Z(c). The difference Z(s)=Z(sc)-Z(c) is the acoustic impedance contribution of the footplate and annular ligament. Z(sc) is mainly determined by the stiffness of the annular ligament at low frequencies, and by the cochlear input impedance Z(c) at higher frequencies. Z(c) is predominately resistive, a minor reactive part at low frequencies is attributed to the stiffness of the round window membrane. Z(s) and Z(c) are equal in magnitude at about 0.4 kHz. Rather close RLC fits have been obtained for all the three impedances, Z(sc), Z(s), and Z(c). The fitted values for the resistive parts of Z(sc) and Z(c) are 62.9 and 58.2 acoustic Gomega, respectively. The relatively small difference, 4.7 Gomega, is attributed to the resistance of the annular ligament. The fitted resistance of Z(s) is somewhat larger, 8.6 Gomega, but is anyway of minor importance relative to the dynamic stiffness of the annular ligament. This stiffness depends on the static pressure difference across the footplate. Each of the averaged Z(sc) corresponds to minimum stiffness. The fitted acoustic compliance is 6.89 x 10(-15) m3/Pa. The acoustic inertance plays a minor role. It is attributed to the mass of the footplate and the co-vibrating liquid in the inner ear, and has a fitted value of 4.7 x 10(5) Pa s2/m3. A sound pressure at the eardrum is equivalent to a larger pressure at the footplate, about 16 dB larger at frequencies below 100 Hz, increasing to about 30 dB at 10 kHz. In the vestibulum at the inner side of the footplate, the sound pressure at 20 Hz is about 20 dB below the equivalent pressure at the outer side. The two pressures approach toward higher frequencies, and above 1 kHz they are nearly equal. PMID- 11008813 TI - An automated procedure for identifying spontaneous otoacoustic emissions. AB - An algorithm is described for objectively identifying and measuring spontaneous otoacoustic emissions (SOAEs) using the spectrum that results from transformation of the acoustic waveform measured in the outer ear canal. Prior to spectral analysis, the rms level is calculated for successive short segments of the waveform and only the weakest 25% of the segments are retained for the spectral analysis [the quietest 150 when using 16k-point fast Fourier transforms (FFTs)]. The resulting initial spectrum is scanned for peaks (potential SOAEs) which are then deleted from the spectrum. New values are estimated for the deleted values using linear extrapolations from frequency ranges on either side of the deleted values. The end result is a smoothed spectrum devoid of all local peaks. The initial spectrum is then compared peak-by-peak with the smoothed spectrum, and those peaks having differences that exceed an objectively determined decision criterion are identified as likely SOAEs. The effects of varying some of the important parameter values of the algorithm are described, and the sensitivity of the procedure is evaluated by measuring the detection rate for a Lorentzian peak of known amplitude added to a spectrum otherwise devoid of SOAEs. PMID- 11008814 TI - On the spectral periodicity of transient-evoked otoacoustic emissions from normal and damaged cochleas. AB - The spectral quasi-periodicity of transient-evoked otoacoustic emissions (TEOAE) is well acknowledged since Zwicker described a preferred spacing of 0.4 bark between consecutive peaks in the spectrum of otoacoustic emissions from normal ears. While there is scarce evidence of any anatomical reason for this regularity, several functional models of the cochlea have predicted that the structure of emission spectra reflects important characteristics of cochlear filters. In an attempt to check such predictions, the average regularity of TEOAE spectra was studied in three groups of human subjects, normally hearing adults, healthy neonates, and adults suffering from noise-induced hearing loss. Significant differences in emission periodicities were found. Around 1 kHz, the preferred spacing was close to 130 Hz in normally hearing adult ears and neonates. In contrast, no clear periodicity was found in the group of damaged ears, even though they had clinically normal pure-tone audiometry below 2 kHz. Around 4 kHz, the preferred spacing was close to 240 Hz in normal adults and neonates, whereas TEOAEs were absent in many impaired ears. A phenomenological model assuming that TEOAEs stem from the responses of a slightly disarrayed bank of highly tuned filters predicts that the filter width would be the same in healthy young adults and neonates. In contrast, ears suffering from high frequency hearing loss could exhibit early damaged filters. The proposed method might provide an objective assessment of parameters otherwise difficult to evaluate, especially in neonatal cochleas. PMID- 11008815 TI - A noninvasive method for estimating acoustic admittance at the tympanic membrane. AB - The acoustic admittance at the tympanic membrane (TM), Y(TM), describes the linear acoustic properties of the ear. Here, a noninvasive measurement procedure is developed for estimating Y(TM) in intact ears. The method consists of (1) measuring the admittance in the ear canal Y(EC) with a commercially available earphone-and-microphone system, and (2) estimating Y(TM) via a uniform-tube approximation of the space between the measurement point and the TM. The dimensions of this space are estimated from Y(EC) via an area-estimation algorithm [Keefe et al., J. Acoust. Soc. Am. 91, 470 (1992)] and measurements made with controlled static pressures in the canal. Measurements in artificial loads are used to test the accuracy of the measurement system and to determine sources of error. For accurate admittance measurements: (1) extension of the microphone tube medially beyond the earphone's port is necessary for frequencies above 2 kHz; (2) the acoustic system must be calibrated in known loads with diameters within 15% of the canal diameter, because the source's output characteristics vary with load diameter. The method is applied to intact ears of anesthetized domestic cats; for frequencies below 5 kHz, the estimated Y(TM) in four ears have features that are similar to those of previous measurements made at the cat TM. Sources of error include nonuniform waves generated at the earphone's narrow port, inaccuracy in estimation of canal dimensions, irregular geometry of the canal, and earphone-microphone cross talk. PMID- 11008816 TI - Tests of some common assumptions of ear-canal acoustics in cats. AB - The accuracy of ear-canal admittance and reflectance as measures of the ear's properties depends on the acoustic effects of the canal. Here, measurements of acoustic admittance at different canal locations in domestic cats are used to test three common assumptions. (1) Can a uniform-tube model of the canal represent spatial variations in admittance? Data from cats support this assumption for frequencies below 3 kHz, where the admittance inferred at the tympanic membrane (TM) based on a uniform-tube model differs by less than 3 dB in magnitude and 0.07 periods in angle from the admittance measured at the TM; for higher frequencies greater differences occur. (2) Do large static air pressures in the canal make the middle ear rigid without affecting the properties of the canal space? The measurements reported indicate that large negative static pressures reduce the low-frequency compliance of the cat middle ear to about 10% of the compliance of the canal air volume. Static displacements of the acoustic probe, TM, and canal walls with static pressure may affect estimates of the canal volume and middle-ear compliance by as much as 15% to 20%. (3) Is the acoustic reflectance magnitude constant with position along the canal? Reflectance data from cat ear canals generally support this idea, except within a frequency region near 0.5 kHz for which there is evidence of energy loss. These results demonstrate that noninvasive measurements in the canal describe middle-ear acoustic properties to within tolerances that depend on the effects of the canal. PMID- 11008817 TI - Perceived continuity and pitch perception. AB - Three experiments investigated the importance of perceived stimulus continuity for the perception of the fundamental frequency (F0) of an unresolved complex tone. The F0 of the complex was 250 Hz and the harmonics were bandpass filtered between 5500 and 7500 Hz. In the first experiment, F0 discrimination was measured for single-burst tones with durations of 20, 40, and 80 ms, and for stimuli containing two 20- or 40-ms tone bursts separated by an 8- or 16-ms gap. For the single-burst conditions, there was a large decrease in threshold as the duration was increased from 20 to 40 ms. However, performance in the gapped conditions was much worse than that for the single-burst condition with the same cumulative duration (e.g., two 20-ms bursts separated by 8 ms produced higher thresholds than one 40-ms burst). Adding a bandpass noise (with the same spectral envelope as the tone) in the gap between the two tone bursts improved performance to the level of the single-burst condition. When the noise was added, the two discrete tone bursts were perceived as one single tone burst interrupted by the noise, and this seemed to facilitate discrimination. In a second experiment, the effects on pitch of an envelope delay (phase shift) of 0.75 periods between two tone bursts separated by an 8-ms gap were investigated. If the gap was silent, the pitch of the pair was unaffected by the phase shift. However, if the gap contained the bandpass noise, the phase shift between the bursts did produce a significant downward shift in the pitch of the pair. Finally, the third experiment showed that presenting a noise before a single 20-ms burst may improve discrimination performance in some listeners, but not sufficiently to account for the results of the first experiment purely in terms of an improvement in the discriminability of the second tone burst in the pair. The experiments suggest that a level decrease between two tone bursts may disrupt or reset a long integration mechanism, decreasing performance. When there is no level decrease between the bursts, the auditory system may assume that the two bursts belong to the same single tone and analyze them together in order to derive F0. PMID- 11008818 TI - The lower limit of pitch as determined by rate discrimination. AB - This paper is concerned with the lower limit of pitch for complex, harmonic sounds, like the notes produced by low-pitched musical instruments. The lower limit of pitch is investigated by measuring rate discrimination thresholds for harmonic tones filtered into 1.2-kHz-wide bands with a lower cutoff frequency, F(c), ranging from 0.2 to 6.4 kHz. When F(c) is below 1 kHz and the harmonics are in cosine phase, rate discrimination threshold exhibits a rapid, tenfold decrease as the repetition rate is increased from 16 to 64 Hz, and over this range, the perceptual quality of the stimuli changes from flutter to pitch. When F(c) is increased above 1 kHz, the slope of the transition from high to low thresholds becomes shallower and occurs at progressively higher rates. A quantitative comparison of the cosine-phase thresholds with subjective estimates of the existence region of pitch from the literature shows that the transition in rate discrimination occurs at approximately the same rate as the lower limit of pitch. The rate discrimination experiment was then repeated with alternating-phase harmonic tones whose envelopes repeat at twice the repetition rate of the waveform. In this case, when F(c) is below 1 kHz, the transition in rate discrimination is shifted downward by almost an octave relative to the transition in the cosine-phase thresholds. The results support the hypothesis that in the low-frequency region, the pitch limit is determined by a temporal mechanism, which analyzes time intervals between peaks in the neural activity pattern. It seems that temporal processing of pitch is limited to time intervals less than 33 ms, corresponding to a pitch limit of about 30 Hz. PMID- 11008819 TI - Characterizing frequency selectivity for envelope fluctuations. AB - Three experimental paradigms were used to specify the auditory system's frequency selectivity for amplitude modulation (AM). In the first experiment, masked threshold patterns were obtained for signal-modulation frequencies of 4, 16, 64, and 256 Hz in the presence of a half-octave-wide modulation masker, both applied to the same noise carrier with a bandwidth ranging from 1 to 4 kHz. In the second experiment, psychophysical tuning curves (PTCs) were obtained for signal modulation frequencies of 16 and 64 Hz imposed on a noise carrier as in the first experiment. In the third experiment, masked thresholds for signal-modulation frequencies of 8, 16, 32, and 64 Hz were obtained according to the "classical" band-widening paradigm, where the bandwidth of the modulation masker ranged from 1/8 to 4 octaves, geometrically centered on the signal frequency. The first two experiments allowed a direct derivation of the shape of the modulation filters while the latter paradigm only provided an indirect estimate of the filter bandwidth. Thresholds from the experiments were predicted on the basis of an envelope power-spectrum model (EPSM) which integrates the envelope power of the modulation masker in the passband of a modulation filter tuned to the signal modulation frequency. The Q-value of second-order bandpass modulation filters was fitted to the masking patterns from the first experiment using a least-squares algorithm. Q-values of about 1 for frequencies up to 64 Hz suggest an even weaker selectivity for modulation than assumed in earlier studies. The same model also accounted reasonably well for the shape of the temporal modulation transfer function (TMTF) obtained for carrier bandwidths in the range from 1 to 6000 Hz. Peripheral filtering and effects of peripheral compression were also investigated using a multi-channel version of the model. Waveform compression did not influence the simulated results. Peripheral bandpass filtering only influenced thresholds for high modulation frequencies when signal information was strongly attenuated by the transfer function of the peripheral filters. PMID- 11008820 TI - The use of visible speech cues for improving auditory detection of spoken sentences. AB - Classic accounts of the benefits of speechreading to speech recognition treat auditory and visual channels as independent sources of information that are integrated fairly early in the speech perception process. The primary question addressed in this study was whether visible movements of the speech articulators could be used to improve the detection of speech in noise, thus demonstrating an influence of speechreading on the ability to detect, rather than recognize, speech. In the first experiment, ten normal-hearing subjects detected the presence of three known spoken sentences in noise under three conditions: auditory-only (A), auditory plus speechreading with a visually matched sentence (AV(M)) and auditory plus speechreading with a visually unmatched sentence (AV(UM). When the speechread sentence matched the target sentence, average detection thresholds improved by about 1.6 dB relative to the auditory condition. However, the amount of threshold reduction varied significantly for the three target sentences (from 0.8 to 2.2 dB). There was no difference in detection thresholds between the AV(UM) condition and the A condition. In a second experiment, the effects of visually matched orthographic stimuli on detection thresholds was examined for the same three target sentences in six subjects who participated in the earlier experiment. When the orthographic stimuli were presented just prior to each trial, average detection thresholds improved by about 0.5 dB relative to the A condition. However, unlike the AV(M) condition, the detection improvement due to orthography was not dependent on the target sentence. Analyses of correlations between area of mouth opening and acoustic envelopes derived from selected spectral regions of each sentence (corresponding to the wide-band speech, and first, second, and third formant regions) suggested that AV(M) threshold reduction may be determined by the degree of auditory-visual temporal coherence, especially between the area of lip opening and the envelope derived from mid- to high-frequency acoustic energy. Taken together, the data (for these sentences at least) suggest that visual cues derived from the dynamic movements of the fact during speech production interact with time-aligned auditory cues to enhance sensitivity in auditory detection. The amount of visual influence depends in part on the degree of correlation between acoustic envelopes and visible movement of the articulators. PMID- 11008821 TI - Effects of frequency and level on auditory stream segregation. AB - This study examined the effect of center frequency and level on the perceptual grouping of rapid tone sequences. The sequence ABA-ABA-...was used, where A and B represent sinusoidal tone bursts (10-ms rise/fall, 80-ms steady state, 20-ms interval between tones) and - represents a silent interval of 120 ms. In experiment 1, tone A was fixed in frequency at 62, 125, 250, 500, 1000, 2000, 4000, 6000, or 8000 Hz. Both tones had a level of approximately 40 dB SL. Tone B started with a frequency well above that of tone A, and its frequency was swept toward that of tone A so that the frequency separation between them decreased in an exponential manner. Subjects were required to indicate when they could no longer hear the tones A and B as two separate streams, but heard only a single stream with a "gallop" rhythm. This changeover point between percepts is called the fission boundary. The separation between tones A and B at the fission boundary was roughly independent of the frequency of tone A when expressed as the difference in number of equivalent rectangular bandwidths (ERBs) between A and B, but varied more with frequency when the difference was expressed in barks or cents. In experiment 2, the center frequency was fixed at 250, 1000, or 4000 Hz, and the level of the A and B tones was 40, 55, 70, or 85 dB SPL. The frequency separation of the A and B tones at the fission boundary tended to increase slightly with increasing level, in a manner consistent with the broadening of the auditory filter with increasing level. The results support the "peripheral channeling" explanation of stream segregation advanced by Hartmann and Johnson [Music Percept. 9, 155-184 (1991)], and indicate that the perception of fusion or fission in alternating-tone sequences depends partly upon the degree of overlap of the excitation patterns evoked by the successive sounds in the cochlea, as assumed in the theory of Beauvois and Meddis [J. Acoust. Soc. Am. 99, 2270-2280 (1996)]. PMID- 11008822 TI - Neural system identification model of human sound localization. AB - This paper examines the role of biological constraints in the human auditory localization process. A psychophysical and neural system modeling approach was undertaken in which performance comparisons between competing models and a human subject explore the relevant biologically plausible "realism constraints." The directional acoustical cues, upon which sound localization is based, were derived from the human subject's head-related transfer functions (HRTFs). Sound stimuli were generated by convolving bandpass noise with the HRTFs and were presented to both the subject and the model. The input stimuli to the model were processed using the Auditory Image Model of cochlear processing. The cochlear data were then analyzed by a time-delay neural network which integrated temporal and spectral information to determine the spatial location of the sound source. The combined cochlear model and neural network provided a system model of the sound localization process. Aspects of humanlike localization performance were qualitatively achieved for broadband and bandpass stimuli when the model architecture incorporated frequency division (i.e., the progressive integration of information across the different frequency channels) and was trained using variable bandwidth and center-frequency sounds. Results indicate that both issues are relevant to human sound localization performance. PMID- 11008823 TI - Complex tactile waveform discrimination. AB - Complex vibrotactile waveforms consisting of two superimposed sinusoids at varying phases were presented to the fingertip, and observers made "same different" judgments. It was found that the low-frequency (10Hz+30Hz) waveforms were discriminable from one another while discrimination of the high-frequency (100Hz+300Hz) vibrations was poor. High-frequency adaptation did not impair discrimination of the low-frequency waveforms, suggesting that the RA channel mediated discrimination. Low-frequency adaptation impaired discrimination of the high-frequency stimuli, suggesting that the RA channel likewise mediated the modest level of performance observed in the absence of an adapting stimulus. The results indicate that this channel encodes complex waveforms temporally. A simple model for low-frequency waveform discrimination is proposed. The results obtained with the high-frequency complex waveforms are compatible with the hypothesis that the PC channel integrates stimulus energy over time. PMID- 11008824 TI - Perceptual calibration of F0 production: evidence from feedback perturbation. AB - Hearing one's own speech is important for language learning and maintenance of accurate articulation. For example, people with postlinguistically acquired deafness often show a gradual deterioration of many aspects of speech production. In this manuscript, data are presented that address the role played by acoustic feedback in the control of voice fundamental frequency (F0). Eighteen subjects produced vowels under a control (normal F0 feedback) and two experimental conditions: F0 shifted up and F0 shifted down. In each experimental condition subjects produced vowels during a training period in which their F0 was slowly shifted without their awareness. Following this exposure to transformed F0, their acoustic feedback was returned to normal. Two effects were observed. Subjects compensated for the change in F0 and showed negative aftereffects. When F0 feedback was returned to normal, the subjects modified their produced F0 in the opposite direction to the shift. The results suggest that fundamental frequency is controlled using auditory feedback and with reference to an internal pitch representation. This is consistent with current work on internal models of speech motor control. PMID- 11008825 TI - Acoustic characteristics of English fricatives. AB - This study constitutes a large-scale comparative analysis of acoustic cues for classification of place of articulation in fricatives. To date, no single metric has been found to classify fricative place of articulation with a high degree of accuracy. This study presents spectral, amplitudinal, and temporal measurements that involve both static properties (spectral peak location, spectral moments, noise duration, normalized amplitude, and F2 onset frequency) and dynamic properties (relative amplitude and locus equations). While all cues (except locus equations) consistently serve to distinguish sibilant from nonsibilant fricatives, the present results indicate that spectral peak location, spectral moments, and both normalized and relative amplitude serve to distinguish all four places of fricative articulation. These findings suggest that these static and dynamic acoustic properties can provide robust and unique information about all four places of articulation, despite variation in speaker, vowel context, and voicing. PMID- 11008826 TI - Intelligibility of bandpass speech: effects of truncation or removal of transition bands. AB - An intelligibility of over 90% was reported for keywords in "everyday" 1/3-octave sentences centered on 1500 Hz and having steep transition band slopes of 100 dB/octave [Warren et al., Percept. Psychophys. 57, 175-182 (1995)]. A subsequent study by Warren and Bashford [J. Acoust. Soc. Am. 106, L47-L52 (1999)] found that it was not the 1/3-octave passband, but the transition bands that were chiefly responsible for this high intelligibility: When the passband and transition bands were segregated using filter slopes of 1000 dB/octave, the isolated passband had an intelligibility score of only 24%, while the pair of transition bands had a score of over 80%. In the present study, experiment 1 examined the distribution of information along the transition bands' slopes by truncation at graded downpoints: Truncation at downpoints of 40 dB or more produced no significant change in intelligibility. Experiment 2 closed the gap separating the transition bands so that their slopes intersected at 1500 Hz. This triangular band had a negligible passband (as defined conventionally by 3-dB downpoints) and an intelligibility score of 60%; truncation at downpoints of 50 dB or more produced no significant change in intelligibility. Experiment 3 determined the intelligibilities of rectangular bands (1000-dB/octave slopes) centered on 1500 Hz. Their bandwidths ranged from 3 to 12 semitones in 1-semitone steps, resulting in intelligibility scores increasing monotonically from 14% to 94%. Calculations based upon experiments 2 and 3 showed that the triangular band truncated at 30-dB downpoints had half the intelligibility of a rectangular band having the same frequency range. PMID- 11008827 TI - The relationship between speech perception and electrode discrimination in cochlear implantees. AB - Speech Intelligibility Index (SII) procedures were used to measure the amount of speech information perceived in five frequency bands (170-570, 570-1170, 1170 1768, 1768-2680, and 2680-5744 Hz) by 15 users of the Cochlear Ltd. CI-22M implant and Spectra-22/SPEAK processor. The speech information perceived was compared to that perceived by normal-hearing listeners. The ability of these subjects to discriminate between stimulation on adjacent electrodes corresponding to each frequency band was also investigated, using a 4IFC procedure with random current level variations of between 0% and 60% of the dynamic range. Relative to normal-hearing listeners, speech information was, on average, significantly more reduced in the four frequency regions between 170 and 2680 Hz than in the region 2680-5744 Hz. There was a significant correlation between electrode discrimination ability (when the random level variation encompassed 20% or more of the dynamic range) and the amount of speech information perceived in the four frequency regions between 170 and 2680 Hz. There was no such correlation in the region 2680-5744 Hz, regardless of the extent of random level variation. These results indicate that speech information in the low to medium frequencies is more difficult for implantees to perceive, that this difficulty is correlated with the difficulty in discriminating electrode place in the presence of random loudness variations, and that fine spectral discrimination may be relatively more important in the vowel-formant regions than in higher frequency regions. PMID- 11008828 TI - In vitro measurement of the frequency-dependent attenuation in cancellous bone between 0.2 and 2 MHz. AB - Our goal was to evaluate the frequency dependence of the ultrasonic attenuation coefficient in cancellous bone. Estimates were obtained in immersion, using a substitution method in the through-transmit mode, by scanning 14 human bone specimens (calcaneus). Measurements were performed with three pairs of focused transducers with a center frequency of 0.5, 1.0, and 2.25 MHz, respectively in order to cover an extended frequency bandwidth (0.2-1.7 MHz). When the experimental attenuation coefficient values were modeled with a nonlinear power fit alpha(f)=alpha0 +alpha(I)f(n), the attenuation coefficient was found to increase as f(1.09+/-0.3) over the measurement bandwidth. However, a substantial variation of the exponent n (0.4-2.2) within specimens and also between specimens was observed. The acoustical parameters were compared to bone mineral density. A highly significant relationship was noted between alpha1 and BMD (r2= 0.75, p< 10(-4)). No correlation was found between n and BMD. Several attenuation mechanisms are discussed as well as the potential impact these results may have in in vivo quantitative measurements. PMID- 11008829 TI - Ultrasound-induced lung hemorrhage is not caused by inertial cavitation. AB - In animal experiments, the pathogenesis of lung hemorrhage due to exposure to clinical diagnostic levels of ultrasound has been attributed to an inertial cavitation mechanism. The purpose of this article is to report the results of two experiments that directly contradict the hypothesis that ultrasound-induced lung hemorrhage is caused by inertial cavitation. Elevated hydrostatic pressure was used to suppress the involvement of inertial cavitation. In experiment one, 160 adult mice were equally divided into two hydrostatic pressure groups (0.1 or 1.1 MPa), and were randomly exposed to pulsed ultrasound (2.8-MHz center frequency, 1 kHz PRF, 1.42-micros pulse duration, 10-s exposure duration). For the two hydrostatic pressure groups (80 mice each), 8 in situ peak rarefactional pressure levels were used that ranged between 2.82 and 11.8 MPa (10 mice/group). No effect of hydrostatic pressure on the probability of hemorrhage was observed. These data lead to the conclusion that lung hemorrhage is not caused by inertial cavitation. Also, the higher hydrostatic pressure enhanced rather than inhibited the impact of ultrasonic pressure on the severity (hemorrhage area, depth, and volume) of lesions. These counterintuitive findings were confirmed in a second experiment using a 2 x 5 factorial design that consisted of two ultrasonic pressure levels and five hydrostatic pressure levels (100 mice, 10 mice/group). If inertial cavitation were the mechanism responsible for lung hemorrhage, then elevated hydrostatic pressures should have resulted in less rather than more tissue damage at each ultrasonic pressure level. This further supports the conclusion that the pathogenesis of ultrasound-induced lung hemorrhage is not caused by inertial cavitation. PMID- 11008830 TI - Acoustic flow perception in cf-bats: extraction of parameters. AB - The narrow-band portions of the echolocation pulses seen in cf-bats are a hypothetical substrate for target localization. This localization could be based on estimates of echo envelope amplitude and carrier frequency together with their derivatives. Evaluation of these parameters is referred to as "acoustic flow" in loose analogy to optic flow. It is assessed whether the requirements for this task may be reconciled with known principles of auditory function. For the evaluation of a single echo, this seems to be the case: auditory filter shapes provide sufficient frequency resolution; at the same time envelopes are preserved well and some noise removal is achieved. Nevertheless, should bats not be endowed with additional capabilities for noise removal, analysis of acoustic flow would be limited to favorable signal-to-noise ratios. Multiple, temporally overlapping echoes are probable in any realistic echolocation scenario. In this case, additional auditory processing steps have to be postulated, which allow simultaneous estimation of multiple carrier frequencies and reduction of demodulation distortions. PMID- 11008831 TI - A mathematical analysis of the peripheral auditory system mechanics in the goldfish (Carassius auratus). AB - The dynamic response of the goldfish peripheral auditory system has been analyzed using lumped-parameter mechanical and fluid system models for the swimbladder, Weberian apparatus, and saccule. The swimbladder is treated as a two degree-of freedom mechanical system consisting of two coupled mass-spring-damper arrangements. The swimbladder is coupled to the Weberian ossicles using a phenomenological analysis of the anterior swimbladder tunica externa which permits both stretching and sliding. Analysis of the saccule features only a single degree of freedom, corresponding to the direction of orientation of the ciliary bundles. Inputs to the saccule consist of the transverse canal fluid motion and the motion of the animal's head (assumed to match the local acoustic particle motion). Mechanical properties required for the system equations were estimated from published literature, direct measurements, and curve fits to experimental data for the motions of the swimbladders. The results indicate that the Weberian apparatus has a significant impact on hearing ability over the entire auditory bandwidth, not just at higher frequencies, and that the saccule functions as a displacement sensor above approximately 300 Hz. PMID- 11008833 TI - A software model to estimate zones of impact on marine mammals around anthropogenic noise. AB - Anthropogenic noise impacts marine mammals in a variety of ways. In order to estimate over which ranges this happens, we first need to understand the propagation of noise through the ocean away from the noise source, and, second, understand the relationship between received noise levels and impact thresholds. A software package combining both aspects is presented. (1) A sound propagation model based on ray theory was developed to calculate received noise levels as a function of range, depth, and frequency. (2) Current knowledge of noise impact thresholds for marine mammals was gathered and included in software routines predicting zones of impact on marine mammals around industrial underwater noise sources. As input parameters, this software package requires the source level and spectrum of the noise of interest; physical oceanography data about the local ocean environment such as bathymetry, bottom and surface loss data, and sound speed profiles; and bioacoustical information about the target species in the form of an audiogram, critical auditory bandwidths, spectra of typical animal vocalizations, reported sound levels of disturbance, and criteria for hearing damage. As output, the software produces data files and plots of the zones of audibility, masking, disturbance, and potential hearing damage around a noise source. PMID- 11008832 TI - Masking in three pinnipeds: underwater, low-frequency critical ratios. AB - Behavioral techniques were used to determine underwater masked hearing thresholds for a northern elephant seal (Mirounga angustirostris), a harbor seal (Phoca vitulina), and a California sea lion (Zalophus californianus). Octave-band white noise maskers were centered at five test frequencies ranging from 200 to 2500 Hz; a slightly wider noise band was used for testing at 100 Hz. Critical ratios were calculated at one masking noise level for each test frequency. Above 200 Hz, critical ratios increased with frequency. This pattern is similar to that observed in most animals tested, and indicates that these pinnipeds lack specializations for detecting low-frequency tonal sounds in noise. However, the individual pinnipeds in this study, particularly the northern elephant seal, detected signals at relatively low signal-to-noise ratios. These results provide a means of estimating zones of auditory masking for pinnipeds exposed to anthropogenic noise sources. PMID- 11008834 TI - Zones of impact around icebreakers affecting beluga whales in the Beaufort Sea. AB - A software model estimating zones of impact on marine mammals around man-made noise [C. Erbe and D. M. Farmer, J. Acoust. Soc. Am. 108, 1327-1331 (2000)] is applied to the case of icebreakers affecting beluga whales in the Beaufort Sea. Two types of noise emitted by the Canadian Coast Guard icebreaker Henry Larsen are analyzed: bubbler system noise and propeller cavitation noise. Effects on beluga whales are modeled both in a deep-water environment and a near-shore environment. The model estimates that the Henry Larsen is audible to beluga whales over ranges of 35-78 km, depending on location. The zone of behavioral disturbance is only slightly smaller. Masking of beluga communication signals is predicted within 14-71-km range. Temporary hearing damage can occur if a beluga stays within 1-4 km of the Henry Larsen for at least 20 min. Bubbler noise impacts over the short ranges quoted; propeller cavitation noise accounts for all the long-range effects. Serious problems can arise in heavily industrialized areas where animals are exposed to ongoing noise and where anthropogenic noise from a variety of sources adds up. PMID- 11008835 TI - Application of a multiple-beam model for lateral vibration analysis of a discretely supported rail at high frequencies AB - In this paper a multiple-beam model of the rail, which allows all the essential deformations caused by the lateral vibration, has been applied for studying the lateral vibration behavior related to the discrete supports. Detailed investigation of the lateral vibration of a discretely supported rail has been carried out using a Green's function matrix and the superposition principle. Good agreement between the predictions and measurement data has been reached in terms of accelerance. PMID- 11008836 TI - Active control design for acoustic radiation using mixed-norm optimization AB - Linear matrix inequalities were applied to design a mixed H2/H(infinity) feedback control compensator for a structural acoustic system. The compensator was designed to minimize the H2 norm of radiation filters while constraining the H(infinity) norm through the control path to be less than unity. A trade-off between minimizing sound power radiated and maximum rms gain in the control path resulted in a reliable and robust means of designing compensators for structural acoustic control. PMID- 11008837 TI - Finding the right cross-correlation peak for locating sounds in multipath environments with a fourth-moment function AB - To locate calling animals in reverberant environments from recordings on widely separated receivers, a fourth-moment "Augmented-Template Correlation Function" (ATCF) helps identify which of many peaks in each cross-correlation function is that corresponding to the difference in travel times for the first arrivals (reference-lag). This peak may not be the largest. The ATCF, by providing an approximate correlation between auto- and cross-correlation functions, can be orders of magnitude more efficient in selecting the reference-lag than the alternative of randomly selecting peaks. The ATCF's efficacy increases with the number of paths and their signal-to-noise ratios. PMID- 11008838 TI - A method for individual identification of echolocation signals in free-ranging finless porpoises carrying data loggers. AB - Echolocation click events of a free-ranging juvenile and an adult finless porpoise (Neophocaena phocaenoides) were recorded with an acoustic data logger. Additionally, dive depth and swim speed of the juvenile were recorded with a behavior data logger. Echoes of echolocation signals from the water surface were clearly detected in shallow dives approximately less than 2 m. The delay time between a surface echo and a direct signal corresponded with the two-way transmission time for the animal's depth, indicating that the signals originated from the animal wearing the data loggers. The finless porpoises produced echolocation signals frequently and were thought to be able to detect their depth by listening to echoes from the water surface. PMID- 11008839 TI - Introduction: the happiness in all our lives. PMID- 11008840 TI - The measurement of happiness. AB - Happiness has been defined either as a broad notion of how one feels about their life in general or as an emotional or affective state. Depending on the way researchers define the concept, there have been variable attempts at measurement. With decades of research, we have a better understanding of how to measure the happiness of others. A combination of quantitative and qualitative methods appears to be most productive. If we assume that individuals with disabilities are made happy in the same way as are people without disabilities, then we have a good idea of how to proceed with practical and policy matters. PMID- 11008841 TI - On the continuous calibration of happiness. AB - The nature of happiness is considered. A model is reviewed in which the affect system is continuously recalibrated regarding a given domain of experiences as a function of the dominant affective tone those experiences have over time. In this view, people who experience repeated negative affect in a domain undergo an adjustment such that they become less demanding about the quality of experience producing happiness; people who experience repeated positive affect in a domain undergo an adjustment such that they become more demanding about the quality of experience required for happiness. Each results in a re-balancing of the system. As objective circumstances vary, the person still experiences both happiness and unhappiness. Implications are considered for large life changes and for individuals with disabilities. PMID- 11008842 TI - From function to felicitude: physical disability and the search for happiness in health services research. AB - Historically, research and practice in physical disability have emphasized the destructive and disabling consequences of illness and injury. More recently, we have begun to measure outcomes related to health, satisfaction, and quality-of life. There has been little systematic attention to the study of happiness, felicitude, but data show that even for many of the people with most notable impairment, levels of happiness are high. The quest for felicitude requires methodological changes and, more significantly, greater professional humility to appreciate that people with both physical and intellectual impairments are able to experience and articulate their own satisfaction, pleasure, and joy. Fortified with this unpretentiousness, we can find happiness and begin to incorporate it as an outcome of clinical and public health programs. PMID- 11008843 TI - Happiness as a treatment goal. AB - Happiness is a concept reflecting general satisfaction with one's life and with oneself, not necessarily congruent with objective indicators. A most important ingredient is a positive self-image, depending, in turn, on positive perception of self as compared to one's peers and to the expectations of important figures in one's life. This requires understanding of personal strengths and constructive acceptance of limitations. Helping individuals develop subjective self satisfaction should be the goal in any treatment program, achieved through direct therapeutic interventions as well as environmental supports (e.g., provision of opportunities for success). It is essential that service providers differentiate between their own values, goals, and administrative requirements and the individual's personal preferences and wishes. PMID- 11008844 TI - Williams syndrome and happiness. AB - Williams syndrome is a genetic disorder resulting in a variety of medical and developmental features, one of which is a frequent outward presentation of substantial happiness. In this paper we describe the unique expression of happiness in people with Williams syndrome, with several anecdotes and a frame by frame conversational analysis. We then discuss this happiness in the context of other dimensions of the impact of Williams syndrome, especially anxiety. We conclude with a discussion of the role of genetics in emotions. PMID- 11008845 TI - Learning about happiness from persons with Down syndrome: feeling the sense of joy and contentment. AB - One well-known stereotype about persons with Down syndrome is that they are relentlessly "happy." Although some individuals with Down syndrome seem to possess an unusual personality aspect that calls out the best in others, this aspect should be viewed as distinct from the traditional stereotype. Many people with Down syndrome have shattered popular expectations of what they might accomplish, society still has lowered expectations for them. In this paper I seek to identify those characteristics that offer genuine and thoughtful joyfulness among these individuals. Happiness is closely aligned with a sense of peace, well being, and/or contentment. A more appropriate contention is the idea that a rare and potent trait holds within it true enrichment and peace among individuals with Down syndrome. PMID- 11008846 TI - Strategic planning for employee happiness: a business goal for human service organizations. AB - Employee happiness can impact substantially on an organization's performance. It can influence employee retention, absenteeism, and work performance. Because of this importance, such happiness is inseparable from the real business of the organization and should be considered a business goal. Implementation and development of the strategic plan associated with this goal becomes the responsibility of a highly placed project team that has as its mission ensuring employee satisfaction. The strategic plan includes procedures that allow management to listen effectively to employees, assessing and responding to their values and needs. In this paper we discuss the workforce and environmental characteristics that are involved planning for employee happiness and the steps in creating an organizational culture in which this can become a business goal. PMID- 11008847 TI - Increasing independent decision-making skills of women with mental retardation in simulated interpersonal situations of abuse. AB - The effectiveness of two decision-making training approaches in increasing independent decision-making skills of 36 women with mild mental retardation in response to hypothetical social interpersonal situations involving abuse was evaluated. Participants were randomly assigned to a control or one of two training conditions (a decision-making training approach that either addressed both cognitive and motivational aspects of decision-making or included only instruction on the cognitive aspect of decision-making). Although both approaches were effective relative to a control condition, the combined cognitive and motivational training approach was superior to the cognitive only training approach. The superiority of this approach was also reflected on a verbally presented generalization task requiring participants to respond to a decision making situation involving abuse from their own perspective and on a locus of control scale that measured perceptions of control. PMID- 11008848 TI - Quantifying the process and outcomes of person-centered planning. AB - Although person-centered planning is a popular approach in the field of developmental disabilities, there has been little systematic assessment of its process and outcomes. To measure person-centered planning, we developed three instruments designed to assess its various aspects. We then constructed variables comprising both a Process and an Outcome Index using a combined rational empirical method. Test-retest reliability and measures of internal consistency appeared adequate. Variable correlations and factor analysis were generally consistent with our conceptualization and resulting item and variable classifications. Practical implications for intervention integrity, program evaluation, and organizational performance are discussed. PMID- 11008849 TI - Mental health of refugees, internally displaced persons and other populations affected by conflict. PMID- 11008850 TI - Juvenile bipolar disorder. AB - OBJECTIVE: Bipolar disorder in children and adolescents is less well studied than bipolar disorder in adults. This review addresses issues related to its underdiagnosis, precursors of bipolarity, comorbidity, natural course and treatment. METHOD: Literature from Medline and other searches, and earlier relevant articles including references from recent review articles on juvenile bipolarity were reviewed. RESULTS: Bipolar disorder in juveniles is underdiagnosed and misdiagnosed on various counts. Few recent studies have reported high rates of comorbid attention deficit and disruptive disorders, prompting some researchers to consider them as probable developmental precursors of juvenile bipolarity. There is also evidence to suggest that some juvenile depression could be pre-bipolar, and that certain temperamental predispositions are probable precursors to bipolarity. Limited data on the natural course and outcome suggest that juvenile bipolar disorder is a highly recurring illness as in adults, and that it is associated with significant functional impairment. The psychopharmacological treatment of juvenile bipolar disorder is remarkably understudied, and treatment is often based on studies of adults. CONCLUSION: There is a need for epidemiological studies of juvenile bipolar disorder. Similarly, there is an urgent need for the methodologically rigorous studies to establish the efficacy of various antimanic drugs. Finally, issues related to comorbidity and temperamental predispositions to juvenile bipolarity need greater clarity, as they may have important treatment and research implications. PMID- 11008851 TI - The prevalence of mental health problems in Rwandan and Burundese refugee camps. AB - OBJECTIVE: We examined the prevalence of mental health problems in refugees living in camps that emerged in Tanzania during the Rwanda crisis that started in 1994. METHOD: Using the 28-item version of the General Health Questionnaire (GHQ 28), we examined two samples: a random sample (n = 854) and a sample of clients of a psychosocial support programme in these camps (n = 23). Sensitivity, specificity and positive- and negative predictive values were estimated for several cut-off scores of the GHQ-28. RESULTS: The prevalence of serious mental health problems was estimated at 50% (SE 12%). When using the GHQ-28 as a screener, a cut-off score of 14 is recommended. CONCLUSION: Given the high prevalence of mental health problems, psychosocial programmes for large refugee populations should aim at strengthening community structures and supporting groups instead of focusing at individuals. The screening capacity of the GHQ-28 could be used to identify mentally vulnerable groups. PMID- 11008852 TI - The 12-month prevalence and risk factors for major depressive episode in Finland: representative sample of 5993 adults. AB - OBJECTIVE: This study reports the 12-month prevalence of major depressive episode and its risk factors in a representative nationwide sample. METHOD: A random sample of non-institutionalized Finnish individuals aged 15-75 years (N = 5993) was interviewed in 1996. Major depressive episode during the last 12 months was assessed using the Short Form of the University of Michigan version of the Composite International Diagnostic Interview (the UM-CIDI Short Form). RESULTS: The population prevalence of major depressive episode was 9.3% [95% CI 8.5,10.0], and the age-adjusted prevalences for females and males were 10.9% [95% CI 9.7,12.0] and 7.2 [95% CI 6.2,8.2], respectively. In logistic regression analyses the factors associated with major depressive episode after adjustment for age were urban residency, smoking, alcohol intoxication and chronic medical conditions. In addition, being single and obese were found to be risk factors for males. CONCLUSION: The female to male risk ratio for major depressive episode was smaller than in many previous studies. The sex-specific risk factor associations warrant further investigation into sex differences in depression. PMID- 11008853 TI - Dissociative disorders in the Sultanate of Oman. AB - OBJECTIVE: To study the clinical manifestations and psychosocial aspects of dissociative (conversion) disorders (DD) in the Sultanate of Oman. METHOD: A retrospective analysis was made of data derived from case records of patients admitted as in-patients to the Sultan Qaboos University Hospital, Oman, over a period of 8 years. RESULTS: A higher prevalence of DD was noted in Oman compared to western countries. The female preponderance was less marked in Oman. The most common presentations were dissociative convulsions, dissociative motor and dissociative trance disorders. Interpersonal difficulties with family and academic problems were the important identified precipitants. CONCLUSION: The study highlights the characteristic clinical and psychosocial features of the dissociative phenomena in Oman. PMID- 11008854 TI - Social phobia treated as a problem in social functioning: a controlled comparison of two behavioural group approaches. AB - OBJECTIVE: Treatments for social phobia result typically in significant anxiety and avoidance reduction; the repercussions in terms of social functioning, however, are not clear. This controlled study compared two approaches designed to improve the social functioning of social phobics. METHOD: Sixty-eight socially phobic patients were randomly assigned to two treatments focused on improving interpersonal relationships either with or without social skills training or a waiting list; 60 completed treatment and 59 a 1-year follow-up. Treatment was administered in small groups, 14 sessions altogether. RESULTS: No clinically meaningful change was observed during the waiting period. A statistically significant and equivalent improvement obtained in both treatment conditions. CONCLUSION: Both treatments resulted in reduced anxiety, avoidance, general psychopathology and better social functioning that maintained over follow-up. Continuing improvement in remission rates was noted; fully 60% of the patients no longer fulfilled criteria for social phobia at the end of 1-year follow-up. PMID- 11008855 TI - Relation of factor-analyzed symptom dimensions of obsessive-compulsive disorder to personality disorders. AB - OBJECTIVE: To assess the relation between symptom dimensions of obsessive compulsive disorder (OCD) and comorbid personality disorders (PDs). METHOD: The scores of 75 OCD outpatients on five previously identified symptom dimensions were entered into multiple regression models as predictors of: 1) the presence of any type of PD; 2) the number of PDs; 3) the presence of any cluster A, B or C PD; and 4) the presence of each individual PD. RESULTS: Twenty-eight patients (37.3%) met criteria for one or more PDs. High scores on the 'Hoarding' dimension were strongly related to the presence of any Axis II diagnosis, and to the number of PDs. Cluster C PDs (especially obsessive-compulsive and avoidant) had the highest partial correlations with 'Hoarding'. These results were independent of OCD symptom severity. CONCLUSION: Previous conflicting findings about the prevalence of certain PDs in OCD might be due in part to differences in the constitution of the particular patient groups studied. PMID- 11008856 TI - Medication adherence in psychosis: predictors and impact on outcome. A 2-year follow-up of first-admitted subjects. AB - OBJECTIVE: To assess the baseline characteristics predicting poor medication adherence following a first admission for psychosis, and the impact of poor medication adherence on outcome. METHOD: First-admitted subjects with psychosis (n = 65) were assessed at 6-month intervals over a 2-year follow-up. Medication adherence was assessed using multiple sources of information. RESULTS: Baseline lower occupational status, alcohol misuse and the intensity of delusional symptoms and suspiciousness predicted poor medication adherence during the 2-year follow-up. Over this period, subjects with poor medication adherence presented more frequently with an episodic course of illness and were more frequently readmitted, especially with regard to involuntary readmission. CONCLUSION: In naturalistic conditions one out of two subjects with psychosis interrupts his/her treatment in the months following his/her first discharge from hospital. Therapeutic programmes aimed at improving medication adherence should be implemented early in the course of psychosis to reduce the deleterious consequences of poor medication adherence on clinical outcome. PMID- 11008857 TI - The relationship between insight and medication adherence in severely mentally ill clients treated in the community. AB - OBJECTIVE: To study the relationship between aspects of insight and medication adherence and elements of psychopathology. METHOD: Clients completed insight and adherence measures in interview; doctors and case managers independently completed ratings of adherence and psychopathology. Other client information was obtained from the clinical file. RESULTS: The subscale structure of the insight scale was well reproduced. Clients endorsed items relating to need for treatment more than items relating to relabelling of symptoms and awareness of illness. Self-reported insight was lower in those clients for whom either the treating doctor or the case manager or both thought the client was non-adherent. Insight was related positively to clinician ratings of depressed mood, and negatively to clinician ratings of personal functioning. CONCLUSION: Our data support the subscale structure of the insight questionnaire used, and previous suggestions that insight is positively related to depressive affect, and inversely related to impaired psychosocial functioning. PMID- 11008858 TI - Swedish universities Scales of Personality (SSP): construction, internal consistency and normative data. AB - OBJECTIVE: A thorough revision of the Karolinska Scales of Personality (KSP) was made by reducing the number of items and improving the psychometric quality as concerns face validity, internal consistency and response differentiation. The revised version was labelled The Swedish universities Scales of Personality (SSP) and now include 91 items divided into 13 scales. METHOD: The SSP were evaluated in a normative, randomly drawn sample (n = 741). All scales were found to be approximately normally distributed. RESULTS: The Cronbach's alpha coefficients ranged from 0.59 to 0.84. The mean inter-item correlations (MIIC) ranged from 0.17 to 0.43. The scale intercorrelation matrix yielded a three-factor solution with Factor 1 reflecting Neuroticism; Factor 2 Aggressiveness; and Factor 3 reflecting Extraversion. CONCLUSION: The original KSP scales were revised, shortened, modernized and psychometrically evaluated. The psychometric properties and the usefulness of the test battery were found to be substantially improved. PMID- 11008859 TI - Prospective evaluation of circulatory levels of catecholamines and serotonin in neuroleptic malignant syndrome. AB - OBJECTIVE: Neuroleptic malignant syndrome (NMS) may be associated with a dysregulation of the catecholaminergic and serotonergic systems. The objective of the present study was to evaluate prospectively the circulatory levels of serotonin (5-HT), epinephrine (E) and dopa in patients suffering from NMS. METHOD: Platelet-poor plasma (PPP) levels of serotonin, epinephrine and dopa in eight NMS patients were measured twice: in the acute state and in the state of remission. RESULTS: PPP dopa concentration was significantly lower in acute NMS state compared to the remission state (P = 0.023). In contrast, PPP E level was significantly higher (P = 0.019) in the acute NMS state and PPP 5-HT concentrations in the acute state tended to be higher than those at remission (P = 0.078). 5-HT/dopa ratio was significantly higher in the acute NMS (P = 0.015). CONCLUSION: These results may reflect reduction in dopaminergic function and increase in adrenergic and serotonergic activity in the acute NMS state. PMID- 11008860 TI - Neuroleptic malignant syndrome during olanzapine and levomepromazine treatment. AB - OBJECTIVE: To date only five reports of neuroleptic malignant syndrome (NMS) related to olanzapine exist. The first case report was published in November 1998. METHOD: We report the case of a 78-year-old woman suffering from chronic schizophrenia who developed a NMS while being treated with olanzapine and levomepromazine. Before this her medication had been unchanged for more than 2 years. RESULTS: When treated with olanzapine and levomepromazine, the patient had a fulminant NMS which was complicated with pneumonia. When the neuroleptic drug treatment was discontinued, the patient recovered. However, when this combination was restarted later due to severe agitation and hallucinations, the symptoms of NMS reappeared. CONCLUSION: This case report shows that the neuroleptic malignant syndrome can occur during olanzapine treatment as well as during treatment with conventional neuroleptics. This syndrome may develop even after a long and stable neuroleptic treatment. PMID- 11008861 TI - Neurological abnormalities in schizophrenia--a 5-year follow-up study: comment on Madsen et al. PMID- 11008862 TI - A simple splinting technique for the mallet finger. PMID- 11008863 TI - Pediatric analgesia and sedation for the management of orthopedic conditions. AB - Orthopedic injuries are common in the emergency department; this is especially true in children. The orthopedist must often supervise sedation and analgesia for the injured child. While the techniques of conscious sedation have improved, the therapeutic index between adequate and excessive sedation is much narrower in children than adults, especially for painful procedures. The purpose of this review is to outline a reasonable approach to sedation and analgesia in the pediatric patient. In addition, current monitoring guidelines are reviewed, as well as the basic pharmacology of the most commonly used drugs. PMID- 11008864 TI - Total ankle replacement: a surgical discussion. Part II. The clinical and surgical experience. PMID- 11008865 TI - The relationship between dementia score and ambulatory level after hip fracture in the elderly. AB - Dementia or mental status has been considered a significant prognosticating factor for the outcome of hip fracture in the elderly; however, the relationship between dementia level and ambulatory level has not been investigated directly. This study was designed to investigate the relationship between ambulatory level and dementia level after hip fracture in the elderly. A retrospective study was performed of 162 hip fracture patients over 65 years old. We measured the dementia score (mini mental state test) of all patients and evaluated their ambulatory status (divided into three groups: dependent, partially dependent, and independent). We then investigated the relationship between ambulatory level and dementia level. Mean dementia score for each group was as follows: dependent group, 6.6 +/- 0.9; partially dependent group, 17.9 +/- 0.8; and independent group, 24.6 +/- 0.4 (+/- SE). There was a significant relationship between ambulatory level and dementia level (P< .0001). PMID- 11008866 TI - Construct stiffness of different fixation methods for supracondylar femoral fractures above total knee prostheses. AB - Supracondylar fracture of the femur after total knee arthroplasty has an estimated frequency of 0.6%-2.5% among total knee recipients and presents an extremely difficult problem when encountered. The goal of this study is to determine the most stable method of fixation of these supracondylar fractures among currently available devices. Synthetic composite femurs with properties similar to human bone were used, and identical, unstable supracondylar fractures were created in each. Osteotomized specimens were placed into four groups of five. Each group was then tested with one of four devices: the Green-Seligson Henry (GSH) intramedullary nail, AO 95 degrees blade plate, dynamic condylar screw and sideplate, and condylar buttress plate. After stabilization with the different types of fixation, the constructs were tested individually for bending stiffness in four modes: flexion, extension, varus, and valgus bending. The stiffest fixation was determined in each of the four bending planes. Resistance to all tested directions was greatest for the condylar screw and sideplate construct. Resistance to flexion (stiffness = 30.96 N/mm), extension (stiffness = 36.36 N/mm), varus (stiffness = 35.46 N/mm), and valgus forces (stiffness = 32.26 N/mm) was highest in the group fixed with the dynamic condylar screw. This may be due to the purchase gained by the large lag screw into the distal femur, or it may be the result of the total rigidity of the implant. Although the femoral samples used in this study do not duplicate the typical osteopenic bone encountered at the site of a total knee arthroplasty, they do allow direct comparison of the fixation devices by removing the variability associated with cadaveric bone samples. PMID- 11008867 TI - Quantitative anatomical study of the posterior interosseous nerve. AB - An anatomic study of the posterior interosseous nerve (PIN) in 20 cadaver upper limbs was performed to measure different segments of the PIN and its relationship with radius and ulna (results given as mean +/- SD). The length of the PIN from radial head to the arcade of Frohse (AF) was 26.5 +/- 1.6 mm and 25.3 +/- 1.1 mm in male and female cadavers, respectively. The length of the PIN from radial head to the PIN exit point from the supinator was 66.7 +/- 4.7 mm and 64.0 +/- 2.5 mm in male and female cadavers, respectively. The overall length of the PIN underlying the supinator muscle was 44.0 +/- 0.5 mm and 37.0 +/- 0.5 mm in male and female cadavers, respectively. The distance between the PIN exit point from the supinator and the radial margin of the radius was 15.0 +/- 0.9 mm and 14.5 +/ 0.9 mm in male and female cadavers, respectively. The distance between the PIN exit point from the supinator and ulnar margin of ulna was 18.2 +/- 0.6 mm and 17.9 +/- 0.7 mm in male and female cadavers, respectively. In 70% (n = 14) of the cadavers, the AF was tendinous and in 30% (n = 6), it was membranous. The length, width, and thickness of AF in males and females, respectively, were 18.6 +/- 1.2 mm / 18.5 +/- 1.3 mm; 2.8 +/- 0.4 mm / 2.5 +/- 0.4 mm; and, 0.8 +/- 0.08 mm / 0.7 +/- 0.07 mm. In all specimens, the PIN exited through the distal supinator muscle by penetrating the muscle. The PIN exit point from the supinator belly was about 11-19 mm from distal border of the latter. The mean distances between PIN exit point from the supinator and the origin of the extensor digitorum communis, abductor pollicis longus, and extensor pollicis longus branches were 7.5 mm, 31 mm, and 58 mm, respectively. PMID- 11008868 TI - Achilles tendon rupture associated with ankle fracture. AB - The case of a 40-year-old man who sustained a medial malleolar fracture with extension of the fracture into the tibial plafond is discussed. Before surgery, the physical examination revealed an Achilles tendon rupture. Surgical treatment to repair the bone and tendon injury was performed. Achilles tendon rupture is not an uncommon injury, but it is rarely associated with a fracture. When a fracture is present, the Achilles tendon injury can be overlooked, which may result in a delay of treatment or residual morbidity. PMID- 11008869 TI - A solitary osteochondroma of the pediatric thoracic spine: a case report and review of the literature. AB - The objective of this study design is to describe the diagnosis and successful treatment of a pediatric patient with an osteochondroma of the thoracic spine. An osteochondroma is a bone tumor that rarely occurs in the thoracic spine, especially in the pediatric population. A simple painless mass may be the only presenting symptom. The laboratory findings are usually nonspecific. Radiographs may be nondiagnostic in certain cases, and computed tomography is the imaging modality of choice. The diagnosis, treatment, and outcome of a pediatric patient with an osteochondroma of the thoracic spine, including a possible genetic predisposition, are reviewed, along with a complete review of the literature. Anteroposterior and lateral plain radiographs illustrated a well-defined solid mass arising from the posterior elements of the tenth thoracic vertebrae. A computed tomography (CT) scan further delineated that the mass arose from the spinous process with no obvious impingement of the nerve roots. After excision of the lesion, gross pathological and histologic evaluation was consistent with an osteochondroma. The use of CT allowed accurate diagnosis of the osteochondroma. This led to appropriate surgical intervention, resulting in definitive treatment. PMID- 11008870 TI - Type II Monteggia lesion with fracture-separation of the distal physis of the radius. AB - We report a rare case of a type II Monteggia lesion with fracture-separation of the distal physis of the radius in a 12-year-old boy and discuss the mechanism of this injury. PMID- 11008871 TI - Contributing factors influencing the functional outcome of floating knee injuries. AB - The purpose of the present study was to retrospectively review the floating knee injuries treated at our institute and to determine various factors, such as severity of soft-tissue or skeletal injuries, site of fractures, and treatment methods that may significantly influence the final functional result in these injuries. Between 1986 and 1996, 65 patients with 66 floating knee injuries were treated in our institution. Among 66 fractures of the femur, 19 (29%) were open. There were 43 open tibial fractures. Fifty cases were Fraser type I floating knee fractures, 7 were type IIa, 2 were type IIb, and 7 were type IIc. In 63 cases (95%), both bones had been surgically stabilized with interlocked nails, Ender pins, plates, screws with/without pinning, or external fixations. Final functional results were evaluated according to Karlstrom and Olerud's criteria. Satisfactory results were rated as cases with excellent or good results. The mean follow-up time was 16.6 months range, (12-50 months). We assessed various factors influencing functional results, including Fraser type, severity of open injury grade (Gustilo) in both fractures, combination of open/closed injuries, fracture types (AO/ASIF type), existence of multiple trauma, neurovascular injuries, polyskeletal trauma, and stabilizing method or operation timing of both fractures. Satisfactory rates in Fraser type I and type II were 64% and 25%, respectively (P= .02). The satisfactory rate in closed, grade I+II, and grade III injuries of the femoral fractures was 53.2%, 81.8%, and 25%, respectively (grade I+II vs. grade III: P < .03). There were no significant correlations between the functional result and the following factors: soft-tissue injuries of the tibia; the fracture pattern of both fractures; the combination of open/closed injuries in each fracture; injury severity score; the existence of neurovascular injuries and double femoral fractures; treatment methods; and operation timing. Severity of damage to the knee joint and open injuries in the thigh were found to be significant factors contributing to the functional outcome in floating knee injuries. PMID- 11008872 TI - Targeting device for pinning finger fractures. AB - An inexpensive targeting system for pinning phalangeal fractures is described. PMID- 11008873 TI - Cervical spine trauma. PMID- 11008874 TI - Semicarbazide-sensitive amine oxidases: widespread occurrence and many names and faces. PMID- 11008875 TI - Semicarbazide-sensitive amine oxidases in heart and bovine serum. AB - In guinea pig dorsal skin the semicarbazide-sensitive amine oxidase (SSAO) is localised in fibroblasts. Fibroblasts in culture lose the ability to express this enzymatic activity with doublings, thus suggesting that the SSAO expression needs some factors which are not present in the 10% bovine serum culture medium. Fresh bovine serum of adult animals contains two SSAO activities, one with high affinity for benzylamine and one with lower affinity. The enzyme with lower affinity for benzylamine was identified as spermine oxidase, the oxidation of [14C]-benzylamine was inhibited by semicarbazide, alpha-aminoguanidine and B24, a specific inhibitor of benzylamine oxidase and spermine oxidase, both SSAO enzymes. The enzymatic activity of bovine serum was partially purified, the kinetic properties and sensitivity to inhibitors studied. A mathematical procedure for the analysis of the kinetics resulting from the activity of two enzymes acting on the same substrate seems to give better results than the methods previously described. PMID- 11008876 TI - On the primary structure of membrane-bound semicarbazide-sensitive amine oxidase (SSAO). AB - Mammalian semicarbazide-sensitive amine oxidase (SSAO) activities are a diverse group of copper dependent enzymes within the classification EC 1.4.3.6. [amine:oxygen oxidoreductase (deaminating) (copper-containing)]. They include plasma amine oxidase and the membrane-bound enzyme. Although soluble plasma SSAO from several species have been successfully purified and cloned, relatively little work has been carried out on the molecular properties of the tissue-bound enzyme. At present there is not conclusive evidence that allows to conclude whether or not plasma SSAO is released from the tissue-bound enzyme. This review focuses upon recent progress made in determining the primary structure of the membrane-bound SSAO. PMID- 11008877 TI - Age-related changes of MAO-A and -B distribution in human and mouse brain. AB - Age-related changes of MAO-A and -B were studied in human and BL/C57 mouse brain areas (substantia nigra, putamen and cerebellum). [3H]Ro41-1049 and [3H]lazabemide were used as selective radioligands to image and quantify MAO-A and MAO-B respectively by enzyme autoradiography. MAO-A binding was higher in mouse, whereas MAO-B binding was higher in human. With aging, mouse MAO-A was significantly reduced between 4 and 8 weeks and remained unchanged until 19 months followed by a slight increase between 19 and 25 months. In contrast, no clear variation was observed in humans between the age of 17-93 years. In most of the structures studied a clear age-related increase in MAO-B was observed beginning in mouse brain at 4 weeks, whereas in human tissue this increase started at the age of 50-60 years. These results show marked differences in the levels and variations of mouse and human MAO-A and -B associated with aging and should be taken into account when extrapolating experimental data from mouse to human. PMID- 11008878 TI - N-methyl(R)salsolinol and a neutral N-methyltransferase as pathogenic factors in Parkinson's disease. AB - The pathogenesis of Parkinson's disease is still an enigma. As an endogenous MPTP like neurotoxin, N-methyl(R)salsolinol was proved to induce parkinsonism in rats and apoptosis in dopaminergic neurons. It is synthesized in the human brain by two enzymes; an (R)salsolinol synthase and an N-methyltransferase, and accumulates in the nigro-striatum in human brains. The activity of a neutral N methyltransferase in the striatum was found to determine the level of MPP+-like 1,2-dimethyl-6,7-dihydroxyisoquinolinium ion, an oxidation product of N methyl(R)salsolinol in the substantia nigra. The activity of this N methyltransferase was found to increase significantly in lymphocytes prepared from parkinsonian patients. In cerebrospinal fluid from untreated parkinsonian patients, N-methyl(R)-salsolinol increases significantly. These results suggest that N-methyl(R)salsolinol and a neutral N-methyltransferase may be endogenous factors in the pathogenesis of Parkinson's disease. PMID- 11008879 TI - Anti-apoptotic function of L-(-)deprenyl (Selegiline) and related compounds. AB - (-)Deprenyl has been proposed to be neuroprotective to dopamine neurons in the parkinsonian brains. To clarify the mechanism, the effects of (-)deprenyl and structurally related compounds on apoptosis induced by a neurotoxin, N-methyl(R) salsolinol, and reactive oxygen species, nitric oxide and peroxynitrite, were examined in dopaminergic SH-SY5Y cells. DNA damage was quantified by the single cell gel electrophoresis (comet) assay. (-)-Deprenyl protected the cells from apoptosis in a dose-dependent way, which required pre-treatment at least for 20 min. The effect was confirmed even after washing out of (-)deprenyl, indicating that (-)-deprenyl initiates the intracellular process to antagonize the apoptotic death program. The studies on the structure-activity relationship reveal that N propargyl residue with hydrophobic structure is essential for the anti-apoptotic function. These results suggest that (-)deprenyl and related compounds may be applicable as neuroprotective agents in neurodegenerative diseases. PMID- 11008880 TI - Molecular interaction between reversible MAO-A inhibitors and the enzyme. Application to aryloxazolidinone, a prototype series. AB - Among the various chemical classes of monoamine oxidase A inhibitors, phenyloxazolidinone represent one of the major series. The purpose of this paper is to review the experimental (X-ray diffraction, NMR, electronic absorption spectroscopy, lipophilicity studies) and theoretical (quantum chemistry, molecular mechanics, molecular dynamics) studies which have led to the description of the mode of interaction between phenyloxazolidinone inhibitors and the MAO-A enzyme. PMID- 11008881 TI - Possible different fates for the hydrogen peroxide produced by rat white adipocyte amine oxidases. AB - Monoamine oxidase (MAO) and benzylamine oxidases (Bz-SSAO) of rat white adipocytes, deaminating benzylamine and tyramine produce hydrogen peroxide at different cellular levels. The peroxide produced by their activity has a very short half-life in adipocyte suspension. The addition of a catalase inhibitor allows for the recovery of the intact peroxide within the first 10-min of its production. Thus, benzylamine and tyramine-dependent peroxide recovery is different, suggesting that the fate of the peroxide produced by the two amine oxidases might be different depending on the cellular site of its production. PMID- 11008882 TI - Unusual pattern of beta-phenylethylamine deamination in the rat heart. AB - The present study was aimed to determine type A and B MAO activities in rat heart and renal cortex homogenates and evaluate the sensitivity of deamination of 3H-5 HT and 14C-beta-PEA to selective MAO-A and MAO-B inhibitors, respectively Ro 41 1049 and lazabemide. Deamination of beta-PEA in the rat heart was not affected (Vmax = 53+/-10 vs 42+/-6 nmol mg protein(-1) h(-1)) by lazabemide (250 nM), but was significantly reduced (Vmax = 10+/-1 nmol mg protein(-1) h(-1)) by Ro 41-1049 (250 nM). Deamination of beta-PEA in the rat heart is a low affinity process (when compared with that in the kidney) with high Km values (244+/-98 vs 18.6+/ 5.8 microM). On the other hand, deamination of 5-HT in the rat heart and renal cortex revealed high Km values, which were similar to those for beta-PEA in the heart. Deamination of beta-PEA (1000 microM) in the rat heart was inhibited in a concentration-dependent manner by Ro 41-1049 with a Ki value of 32 nM (22, 48; 95% confidence limits), but not by the selective MAO-B inhibitor lazabemide (up to 500 nM). Inhibition of 5-HT (1000 microM) deamination in the rat heart by Ro 41-1049 was also a concentration-dependent process with a Ki value of 21 (16, 26) nM. Deamination of 5-HT (1000 microM) in the rat renal cortex, was inhibited in a concentration-dependent manner by Ro 41-1049 with a Ki value of 12 (8, 17) nM. Deamination of beta-PEA in the renal cortex was inhibited by lazabemide with a Ki of 5 (3, 7) nM. In the rat heart, in contrast to that in the renal cortex, the specific MAO-B substrate beta-PEA is deaminated by a form of MAO which most probably corresponds to MAO-A. PMID- 11008883 TI - Structural approach of human MAO-A using fold recognition (threading) techniques. AB - The major goal of the present work is to further approach the structure of human monoamine oxidase A (MAO-A). A first partial three-dimensional model of human MAO A has already been established using secondary structure predictions and fold recognition methods [Wouters and Baudoux, 1998]. In this modeled structure, a segment of the sequence (residues 369-393) located near the covalent linkage to the essential flavin cofactor, and potentially involved in the structure of the active site of the protein, could not be modeled. We here propose a possible fold for that segment, based on threading techniques. The identification of regions of the protein potentially involved in its dimerization was also undertaken by studying hydrophobic areas present at the surface of the structure. PMID- 11008884 TI - Steroid binding by antibodies and artificial receptors: exploration of theoretical methods to determine the origins of binding affinities and specificities. AB - Binding mode calculations for complexes between an artificial paracyclophane receptor and digoxins, cholic acids as well as cortisone steroids show encapsulation of different ring combinations. Docking experiments were performed between the 26-10 antibody and digoxins. Coordination affinity arises from hydrophobic desolvation and van der Waals interactions rather than from hydrogen bonds. The specificity and affinity arises mainly from shape complementarity. Computed binding free energies and Kohonen neural network computations both point to physicochemical and structural similarities of natural antibodies and artificial receptors. PMID- 11008885 TI - Localization and quantification of hydrophobicity: the molecular free energy density (MolFESD) concept and its application to sweetness recognition. AB - A method for the localization, the quantification, and the analysis of hydrophobicity of a molecule or a molecular fragment is presented. It is shown that the free energy of solvation for a molecule or the transfer free energy from one solvent to another can be represented by a surface integral of a scalar quantity, the molecular free energy surface density (MolFESD), over the solvent accessible surface of that molecule. This MolFESD concept is based on a model approach where the solvent molecules are considered to be small in comparison to the solute molecule, and the solvent can be represented by a continuous medium with a given dielectric constant. The transfer energy surface density for a 1 octanol/water system is empirically determined employing a set of atomic increment contributions and distance dependent membership functions measuring the contribution of the increments to the surface value of the MolFESD. The MolFESD concept can be well used for the quantification of the purely hydrophobic contribution to the binding constants of molecule-receptor complexes. This is demonstrated with the sweeteners sucrose and sucralose and various halogen derivatives. Therein the relative sweetness, which is assumed to be proportional to the binding constant, nicely correlates to the surface integral over the positive, hydrophobic part of the MolFESD, indicating that the sweetness receptor can be characterized by a highly flexible hydrophobic pocket instead of a localized binding site. PMID- 11008887 TI - Hydration of beta-cyclodextrin: a molecular dynamics simulation study. AB - We study by molecular dynamics simulations the hydration of beta-cyclodextrin. Our simulations show that within these barrel-shaped molecules hydrophobicity dominates, while at the top and bottom sides of the barrel interactions with water are mostly hydrophilic in nature. These results agree with crystallographic data at 120 K and, in particular, with the spontaneous hydration process of a cyclodextrin crystal in wet atmosphere. The predicted structure of the hydration shells is discussed and compared with previous molecular mechanics calculations which report an overall hydrophobic behavior. Moreover, the temperature dependence of the hydration process is discussed. PMID- 11008888 TI - Classical QSAR and comparative molecular field analyses of the host-guest interaction of organic molecules with cyclodextrins. AB - The application of classical QSAR and molecular modeling analysis using Comparative Molecular Field Analysis (CoMFA) to the complexation of some natural and modified cyclodextrins (CDs) with guest molecules was examined. For 1:1 complexation systems between natural beta-CD, modified alpha-, beta-, and gamma CD that bear one p-(dimethylamino)benzoyl (DMAB) moiety (DMAB-alpha-, beta-, and gamma-CDs) and guest molecules of widely varying chemical structures and properties, the binding constants of the complexes were successfully fitted using multiple linear regression (MLR) with hydrophobic descriptor log P (the partition coefficient between 1-octanol and water phases) and molecular connectivity indices. A non-linear dependency of binding constants on the zero-th and/or first order molecular connectivity index as a measure of size becomes apparent. The modeling performance of the CoMFA models with steric/electrostatic fields to DMAB alpha- and beta-CD systems was comparable to those of MLR models. However, statistically significant CoMFA models for gamma-CD systems which have higher conformational flexibility of the ring could not be obtained. The CoMFA models obtained for DMAB-alpha- and beta-CD systems showed that the predominant effects were steric for the DMAB-alpha-CD system and electrostatic for the DMAB-beta-CD system, respectively. PMID- 11008886 TI - 3D-QSAR using 'multiconformer' alignment: the use of HASL in the analysis of 5 HT1A thienopyrimidinone ligands. AB - The observed 5-HT1A and alpha1-adrenergic receptor (alpha1-AR) receptor binding properties of a series of 23 thienopyrimidinones were used to develop HASL 3D QSAR models. A single, low energy conformer of the most active analogue in the series, which was consistent with NMR structural studies, was chosen as a template molecule. Alignments of all the molecules to the template were provided by an Amber/MM2 superposition force field. In this manner, each molecule was represented by five separate low energy conformers which were subsequently used in the generation of HASL 3D-QSAR models. Models derived from multiple conformers were found to exhibit enhanced predictivity compared to models based on single, low energy conformers. In addition, the use of contour imaging of HASL multi conformer model interactions was found to lead to a more consistent interpretation of those molecular features most significant for 5-HT1A receptor binding. PMID- 11008889 TI - Indices of differences of path lengths: novel topological descriptors derived from electronic interferences in graphs. AB - Novel topological descriptors, namely indices of differences of path lengths (DPs), are deduced from the physical model of wave interferences. Two electrons, moving through a circuit graph within a diffraction experiment, interfere in a given vertex of the graph. It is demonstrated that the overall sum of the inverse of the squares of the differences of topological distances between all pairs of vertices of the graph is a measure of the mean global kinetic energy of the electrons which are able to produce a constructive interference. New topological indices, namely indices of differences of path lengths are thus introduced as derived from such a diffraction pattern. These indices, according to the above expressed, should be a measure of the electron mobility within the molecule. As a consequence, a good prediction is to be expected for properties related to such mobility, such as resonance energy in aromatic hydrocarbons. Our results confirm that in fact, the resonance energies are well predicted by this means. Moreover, the new indices demonstrate to be very useful in the evaluation of biological properties such as antibacterial activities of a wide set of heterogeneous compounds. PMID- 11008891 TI - Primary non familial amyloidosis of the vitreous and subretinal amyloid bodies. AB - We report a case of primary non familial amyloidosis of the vitreous and subretinal amyloid bodies. Pars plana vitrectomy and subretinal numerable clusters of cystic-like glistering material were extracted. The diagnosis was confirmed only by histological findings. PMID- 11008890 TI - Conformational properties of amphotericin B amide derivatives--impact on selective toxicity. AB - Even though it is highly toxic, Amphotericin B (AmB), an amphipathic polyene macrolide antibiotic, is used in the treatment of severe systemic fungal infections as a life-saving drug. To examine the influence of conformational factors on selective toxicity of these compounds, we have investigated the conformational properties of five AmB amide derivatives. It was found that the extended conformation with torsional angles (phi,psi)=(290 degrees,180 degrees) is a common minimum of the potential energy surfaces (PES) of unsubstituted AmB and its amide derivatives. The extended conformation of the studied compounds allows for the formation of an intermolecular hydrogen bond network between adjacent antibiotic molecules in the open channel configuration. Therefore, the extended conformation is expected to be the dominant conformer in an open AmB (or its amide derivatives) membrane channel. The derivative compounds for calculations were chosen according to their selective toxicity compared to AmB and they had a wide range of selective toxicity. Except for two AmB derivatives, the PES maps of the derivatives reveal that the molecules can coexist in more than one conformer. Taking into account the cumulative conclusions drawn from the earlier MD simulation studies of AmB membrane channel, the results of the potential energy surface maps, and the physical considerations of the molecular structures, we hypothesize a new model of structure-selective toxicity of AmB derivatives. In this proposed model the presence of the extended conformation as the only well defined global conformer for AmB derivatives is taken as the indicator of their higher selective toxicity. This model successfully explains our results. To further test our model, we also investigated an AmB derivative whose selective toxicity has not been experimentally measured before. Our prediction for the selective toxicity of this compound can be tested in experiments to validate or invalidate the proposed model. PMID- 11008892 TI - Changes in optic nerve head blood flow and retrobular hemodynamics following calcium-channel blocker treatment of normal-tension glaucoma. AB - BACKGROUND: Because calcium channel blockers reduce vascular resistance, they may have a clinical application in the treatment of normal-tension glaucoma (NTG). This study investigates changes in both the optic disc blood flow and the hemodynamics of retrobulbar vessels in NTG patients after the systemic administration of a calcium channel blocker. METHODS: Twelve eyes of 12 NTG patients (mean age 57.6 +/- 15.3 years) were examined before and after a 4-week treatment with 2 mg b.i.d. oral nilvadipine, an L-typc calcium channel blocker. By scanning laser-Doppler flowmetry (SLDF), we obtained the velocity, flow, and volume from within a 10 x 10 pixel window placed on the temporal rim region of the optic disc perfusion map. By ultrasound color Doppler imaging (CDI), we measured the peak systolic velocity (PSV) and the end diastolic velocity (EDV) of the ophthalmic artery (OA), central retinal artery (CRA), nasal posterior ciliary artery (NPCA), and temporal posterior ciliary artery (TPCA). We then calculated a resistance index (RI) for each vessel. RESULTS: After treatment, the flow and velocity of the optic disc blood flow significantly increased (P < 0.05). Nilvadipine also significantly reduced RIs of the CRA, NPCA, and TPCA (P < 0.05), and increased both the PSV of the NPCA and the EDVs of the CRA, NPCA, and TPCA. The percent change in velocity correlated significantly with the percent changes of the CRA RI and NPCA RI. CONCLUSIONS: Oral nilvadipine appears to reduce orbital vascular resistance, which consequently increases the optic disc blood flow. PMID- 11008893 TI - Orbital color Doppler imaging of optic nerve tumors. AB - OBJECTIVE: To report changes in retinal arterial and venous blood flow pattern in two patients with tumors involving the entire optic nerve. METHODS: Retrospective review of one patient with clinical and neuroimaging characteristics typical of bilateral optic nerve gliomas and one patient with a probable meningioma of the left optic nerve sheath. RESULTS: The optic nerve glioma patient had reduced peak systolic velocity of central retinal arteries bilaterally, while the patient with an optic nerve sheath meningioma had relatively low central retinal artery flow velocity and intermittent blood flow in the central retinal vein on the affected side. CONCLUSIONS: Reduced retinal arterial flow velocities in the setting of optic nerve gliomas may correlate with the presence of optic nerve disease. Phasic blood flow in the central retinal vein with optic nerve sheath meningioma may be the reason that some patients with this tumor develop retinal choroidal venous anastomoses. PMID- 11008894 TI - Perfluorohexyloctane as a long-term vitreous tamponade in the experimental animal. Experimental perfluorohexyloctane substitution. AB - BACKGROUND: Perfluorocarbon liquids are established tools for intraoperative hydrokinetic retinal manipulation. Because of their high specific gravity, however, they may well cause mechanical damage to the retina when applied as a long-term vitreous substitute. Perfluorohexyloctane, a semi-fluorinated liquid fluorocarbon of low specific gravity (1.35 g/cm3), was developed as a long-term vitreous substitute. The current study was performed to investigate intraocular tolerance to perfluorohexyloctane as a long-term vitreous substitute in the experimental animal. METHODS: 34 vitrectomised eyes of pigmented rabbits underwent intravitreal injection of 1.0-1.2 ml perfluorohexyloctane or balanced salt solution. In 5 eyes the anterior chamber was filled. During the follow-up period of 3 months, the eyes were examined by slit lamp biomicroscopy, by fluorescein angiography and by electroretinography. The eyes were then enucleated and processed for light- and electron microscopy. RESULTS: Perfluorohexyloctane depicted dispersion beginning between the first and third week. Over a period of 9 weeks no toxic effect on retina, lens and cornea was noticed. At 14 weeks ERG showed a slight decrease in amplitude and early morphological changes in the retina. CONCLUSION: Our findings suggest that perfluorohexyloctane is tolerated in the rabbit eye for 9 weeks. Since avascular rabbit retina is more susceptible to mechanical and toxic damage than vascularized primate retina, we suggest proceeding from here, and investigating tolerance and quality of tamponade of perfluorohexyl octane in man for up to 9 weeks. PMID- 11008895 TI - Fludrocortisone in the treatment of systemic hypotension in primary open-angle glaucoma patients. AB - BACKGROUND: Fludrocortisone is a potent mineralocorticoid, which has no known direct vasoactive properties, and is the first-line drug for treatment of orthostatic hypotension. The present study evaluated the systemic hemodynamic effects of fludrocortisone treatment in glaucoma patients. PATIENTS AND METHODS: A retrospective analysis of the charts of glaucoma patients of the University Eye Clinic Basel was performed. Twenty-two patients with open-angle glaucoma under treatment with fludrocortisone were selected. The selected patients had one 24-h blood pressure recording immediately prior to treatment with fludrocortisone and one recording at least 2 months after starting the treatment. Parallel to blood pressure recordings, diurnal intraocular tension curve recordings and visual field testings were carried out. In addition, twelve patients also had nail-fold video-capillaroscopy. RESULTS: IOP and visual fields remained stable. The average values for all systemic blood-pressure readings showed an improvement in the follow-up compared to primary examination. Mean (+/- SD) night-to-day ratio ('nocturnal dips') of systolic, diastolic and mean arterial blood pressure decreased from 13.6 +/- 4.3%, 16.9 +/- 5.2% and 15.9 +/- 3.5%, respectively, to 9.9 +/- 5.9% (p = 0.01), 13.2 +/- 4.3% (p = 0.044) and 11.7 +/- 3.9% (p = 0.0004). Baseline capillary blood-flow velocity increased and capillary blood flow standstill time after cold provocation decreased significantly under fludrocortisone therapy. CONCLUSION: Hemodynamic parameters show a tendency towards improvement in a magnitude which might be of clinical relevance. PMID- 11008896 TI - Contrast sensitivity in patients recovered from central serous chorioretinopathy. AB - OBJECTIVE: To study contrast sensitivity in patients who have recovered from central serous chorioretinopathy (CSC). PATIENTS AND METHODS: Thirty-one patients who had recovered from CSC were examined with the Vistech and Pelli-Robson contrast sensitivity charts. The time from the onset of the active disease varied from 10 to 166 months (mean 60.4 +/- 42.0, SD). The visual acuity was 1.0 (logMar 0) or better. RESULTS: Contrast sensitivity of the affected eyes was significantly worse in the intermediate spatial frequencies of 3 and 6 cycles per degree (cpd) in the Vistech test compared to the fellow eyes (p = 0.032, 0.013, respectively). Contrast sensitivity of the affected eyes was significantly worse in all 5 spatial frequencies of the Vistech test and in the Pelli-Robson test compared to age-matched normal eyes (p = 0.006, 0.000, 0.000, 0.018, 0.000, 0.000, respectively). Contrast sensitivity of the fellow eyes was significantly worse in the spatial frequencies of 3 and 18 cpd in the Vistech test and in the Pelli-Robson test compared to age-matched normal eyes (p = 0.020, 0.019, 0.000, respectively). CONCLUSIONS: Contrast sensitivity does not seem to recover in all eyes after CSC even if the visual acuity has returned to normal. Therefore, contrast sensitivity testing is recommended for the patients complaining of visual impairment in spite of good visual acuity. PMID- 11008897 TI - Electrocardiographic changes during subconjunctival injections. AB - Electrocardiographic changes were investigated in 53 patients during subconjunctival drug injections. Decrease in heart-rate of more than 10% or arrhythmia was observed in 25 (47.2%) of the patients, who were considered to have a positive oculocardiac reflex. In the control group of 35 patients, only 3 (8.6%) were found to have a significant decrease in heart rate during blood sampling from antecubital vein (p = 0.000). Young age was a significant risk factor for oculocardiac reflex during injections (p = 0.01). Patient gender, verbal pain score, satiety, experience of periocular injections, injected drug type were not found to be risk factors. All patients with oculocardiac reflex were asymptomatic; however, it is still wise to have emergency supplies ready for any circulatory derangement during subconjunctival injections. PMID- 11008898 TI - Ophthalmologic outcome of transvenous embolization of spontaneous carotid cavernous fistulas: a preliminary report. AB - We give a preliminary report of the ophthalmologic outcome of four patients with transvenous embolization studied for their spontaneous carotid-cavernous fistulas. One of them is a direct type while three of them are indirect dural shunts. In indirect dural shunts, traditional transarterial embolization rarely achieves a complete clinical cure in a short period of time. All cases had an ophthalmologic disturbance justified for endovascular intervention. We performed catheterization, and subsequently embolization with Guglielmi Detachable coils, to the cavernous sinus via the femoral vein and inferior petrosal sinus/superior ophthalmic vein. All four patients achieved clinical and angiographic improvement with a follow-up period range from two to sixteen months except for one patient who had residual bilateral sixth nerve palsy. The transvenous approach offers an effective and safe alternative for the management of spontaneous carotid cavernous fistula. PMID- 11008899 TI - Visual outcomes after cataract surgery and cataract surgical coverage in India. AB - Visual outcomes of 2369 cataract operated persons(3655 eyes) across seven major Indian states were assessed in 1998. This is the largest ever study over the past decade in the country. 9.54 per cent of the examined population had undergone operation for cataract in one or both eyes. Intra-capsular cataract extraction was the commonest surgical modality adopted (91.62%). Intraocular lens implants resulted in better visual outcomes with 71.4 per cent of such patients achieving a good visual outcome (> or = 6/18 in operated eyes). Visual outcome was also good if ICCE operated patients were provided good quality aphakic spectacles. There were no gender differentials in surgical uptake rates. The risk of poor postsurgical visual outcome was strongly associated by type of surgery (Adjusted OR for ICCE-2.78; 95% CI: 1.41-5.49) and the non-availability or poor quality aphakic spectacles (Adjusted OR for poor/no spectacles 4.59-95% CI: 3.53-5.97). Duration since surgery and the source of surgery did not influence visual outcomes. Half the cataract blind remained unoperated in the study population. PMID- 11008901 TI - Epidemiological research at the American Cancer Society. PMID- 11008900 TI - Visual outcome and complications after posterior capsule rupture during phacoemulsification surgery. AB - PURPOSE: To determine the visual outcome and incidence of complications after posterior capsule rupture during phacoemulsification surgery. METHODS: Forty-four eyes of 43 patients which sustained posterior capsule rupture or zonulysis during phacoemulsification surgery between April 1993 and May 1996 were retrospectively studied. Eyes With pseudoexfoliation syndrome, traumatic cataract and myopia > 6 dioptres were excluded from the study. The mean postoperative follow-up period was 26 months (range 3 to 47 months). RESULTS: Forty-one eyes had posterior capsule rupture and 3 eyes had zonulysis. Of these 44 eyes, 41 eyes had vitreous loss and anterior vitrectomy at the time of surgery or subsequently. Excluding 2 eyes with pre-existing conditions that precluded good vision, 36 out of 42 eyes (85.7%) achieved spectacle corrected visual acuity of 6/12 or better post operatively. Of all 42 eyes, 20 had anterior chamber intraocular lens (ACIOL) implantation while the others had posterior chamber intraocular lens (PCIOL) implantation. For eyes with ACIOL, 17 out of 20 eyes (85.0%) had best corrected visual acuity of 6/12 or better. For eyes with PCIOL, 19 out of 22 eyes (86.4%) had best corrected visual acuity of 6/12 or better. CONCLUSIONS: Phacoemulsification surgery, even if complicated by posterior capsule rupture or zonulysis, is compatible with good visual outcome. This is provided prompt attention is paid to the management of complications. PMID- 11008902 TI - Cancer and Mediterranean dietary traditions. AB - The incidence of cancer overall in Mediterranean countries is lower than in Scandinavian countries, the United Kingdom, and the United States. This is mostly accounted for by the lower incidence among Mediterranean countries of cancer of the large bowel, breast, endometrium, and prostate. These forms of cancer have been linked to dietary factors, particularly low consumption of vegetables and fruit, and to a certain extent, high consumption of meat. The traditional Mediterranean diet is characterized by high consumption of foods of plant origin, relatively low consumption of red meat, and high consumption of olive oil, which in several studies has been reported to be more beneficial against cancer than other forms of added lipids. By taking into account the established or presumed nutritional causation of major forms of cancer and the composition of the traditional Mediterranean diet, estimates can be derived concerning the fraction of cancer occurrence in highly developed Western countries that could be attributed to their diets in comparison with the healthy traditional Mediterranean diet. Although estimates can only be crude, it can be calculated that up to 25% of the incidence of colorectal cancer, approximately 15% of the incidence of breast cancer, and approximately 10% of the incidence of prostate, pancreas, and endometrial cancer could be prevented if the populations of highly developed Western countries could shift to the traditional healthy Mediterranean diet. PMID- 11008903 TI - Prostate cancer risk in relation to anthropometry and physical activity: the National Health and Nutrition Examination Survey I Epidemiological Follow-Up Study. AB - We studied the relationship of prostate cancer to anthropometry and self-reported physical activity among 5377 African-American and Caucasian participants in the National Health and Nutrition Examination Survey I cohort. The cohort was first examined between 1971 and 1975 and then followed prospectively through the Epidemiologic Follow-up Study in 1982-1984, 1986, 1987, and 1992. Men who reported low levels of nonrecreational physical activity had increased risk of prostate cancer compared with very active men. These findings were unchanged after adjustment for potential confounders and were stronger for African Americans (relative risk, 3.7; 95% confidence interval, 1.7-8.4) than for Caucasians (relative risk, 1.7; confidence interval, 0.8-2.3). Lower levels of recreational activity were weakly associated with increased prostate cancer risk among African-Americans but not among Caucasians. Prostate cancer risk was unrelated to a variety of anthropometric variables. These results suggest that inactive men are at increased risk of prostate cancer. PMID- 11008904 TI - Serum selenium and subsequent risk of prostate cancer. AB - It is suspected that selenium is protective against prostate cancer. To test this hypothesis, we conducted a nested case-control study in a cohort of 9345 Japanese American men examined between 1971 and 1977. At the time of examination, a blood specimen was obtained, and the serum was frozen. After a surveillance period of more than 20 years, 249 tissue-confirmed incident cases of prostate cancer were identified. Their stored sera and those of 249 matched controls were measured for selenium levels. Odds ratios for prostate cancer, based on quartiles of serum selenium levels, were determined using the General Estimating Equations approach. The multivariate odds ratio for the highest quartile was 0.5 (95% confidence interval, 0.3-0.9) with a two-sided P for trend of 0.02. The inverse association was more notable for cases with advanced disease and for cases diagnosed 5-15 years after phlebotomy. However, the association was mainly present in current or past cigarette smokers rather than nonsmokers, which leads to caution in the interpretation of the results. PMID- 11008905 TI - Gene-covariate interaction between dysplastic nevi and the CDKN2A gene in American melanoma-prone families. AB - The CDKN2A gene has been implicated in cutaneous malignant melanoma pathogenesis. Although CDKN2A mutations confer substantial risk for melanoma, clinicoepidemiological covariates including dysplastic nevi (DN), total nevi, and solar injury also enhance melanoma risk. To examine the relationship between CDKN2A and these three risk factors, we conducted combined segregation/linkage analysis using the class D regressive logistic model, as implemented in the computer program REGRESS. Genetic and covariate data were collected on 20 American melanoma-prone families, 13 of which had cosegregating CDKN2A mutations. Two types of analyses were conducted. The missing-indicator method used a missing value indicator, set to 1 for unknown and 0 for known covariate status, and a second variable set to 1 for exposed and 0 for unexposed or unknown. The second method, complete-cases method, coded subjects with missing covariates as unknown for the affection status. The results for both analyses were very similar. Overall, there was a significant improvement in the likelihood when DN, total nevi or both covariates were added to the base model, which included dominant transmission of the CDKN2A gene and a linear increase of risk with the logarithm of age on the logit scale. In contrast, inclusion of solar injury did not significantly improve the likelihood for the base model. Significant evidence for a gene-covariate interaction was detected between DN and CDKN2A when DN was the only covariate in the model (missing-indicator method or complete-cases method) or when both DN and total nevi were in the model (complete-cases method only). Interestingly, in both methods, the odds ratio (OR) for DN was greater in subjects without mutations (OR, 20.1; 95% confidence interval, 4.8-92.8) versus those with CDKN2A mutations (OR, 3.3; 95% confidence interval, 1.1-10.0; complete cases method). The CDKN2A-DN interaction illustrates the complex etiology of melanoma and needs to be confirmed in a larger sample of families. PMID- 11008906 TI - Empirical Bayes adjustments for multiple results in hypothesis-generating or surveillance studies. AB - Traditional methods of adjustment for multiple comparisons (e.g., Bonferroni adjustments) have fallen into disuse in epidemiological studies. However, alternative kinds of adjustment for data with multiple comparisons may sometimes be advisable. When a large number of comparisons are made, and when there is a high cost to investigating false positive leads, empirical or semi-Bayes adjustments may help in the selection of the most promising leads. Here we offer an example of such adjustments in a large surveillance data set of occupation and cancer in Nordic countries, in which we used empirical Bayes (EB) adjustments to evaluate standardized incidence ratios (SIRs) for cancer and occupation among craftsmen and laborers. For men, there were 642 SIRs, of which 138 (21%) had a P < 0.05 (13% positive with SIR > 1.0 and 8% negative with SIR < or = 1.0) when testing the null hypothesis of no cancer/occupation association; some of these were probably due to confounding by nonoccupational risk factors (e.g., smoking). After EB adjustments, there were 95 (15%) SIRs with P < 0.05 (10% positive and 5% negative). For women, there were 373 SIRs, of which 37 (10%) had P < 0.05 before adjustment (6% positive and 4% negative) and 13 (3%) had P < 0.05 after adjustment (2% positive and 1% negative). Several known associations were confirmed after EB adjustment (e.g., pleural cancer among plumbers, original SIR 3.2 (95% confidence interval, 2.5-4.1), adjusted SIR 2.0 (95% confidence interval, 1.6-2.4). EB can produce more accurate estimates of relative risk by shrinking imprecise outliers toward the mean, which may reduce the number of false positives otherwise flagged for further investigation. For example, liver cancer among chimney sweepers was reduced from an original SIR of 2.2 (range, 1.1 4.4) to an adjusted SIR of 1.1 (range, 0.9-1.4). A potentially important future application for EB is studies of gene-environment-disease interactions, in which hundreds of polymorphisms may be evaluated with dozens of environmental risk factors in large cohort studies, producing thousands of associations. PMID- 11008907 TI - N-Acetyltransferase-2 genetic polymorphism, well-done meat intake, and breast cancer risk among postmenopausal women. AB - Heterocyclic amines found in well-done meat require host-mediated metabolic activation before initiating DNA mutations and tumors in target organs. Polymorphic N-acetyltransferase-2 (NAT2) catalyzes the activation of heterocyclic amines via O-acetylation, suggesting that NAT2 genotypes with high O acetyltransferase activity (rapid/intermediate acetylator phenotype) increase the risk of breast cancer in women who consume well-done meat. To test this hypothesis, DNA samples and information on diet and other breast cancer risk factors were obtained from a nested case-control study of postmenopausal women. Twenty-seven NAT2 genotypes were determined and assigned to rapid, intermediate, or slow acetylator groups based on published characterizations of recombinant NAT2 allozymes. NAT2 genotype alone was not associated with breast cancer risk. A significant dose-response relationship was observed between breast cancer risk and consumption of well-done meat among women with the rapid/intermediate NAT2 genotype (trend test, P = 0.003) that was not evident among women with the slow acetylator genotype (trend test, P = 0.22). These results suggest an interaction between NAT2 genotype and meat doneness, although a test for multiplicative interaction was not statistically significant (P = 0.06). Among women with the rapid/intermediate NAT2 genotype, consumption of well-done meat was associated with a nearly 8-fold (odds ratio, 7.6; 95% confidence interval, 1.1-50.4) elevated breast cancer risk compared with those consuming rare or medium-done meats. These results are consistent with a role for O-acetylation in the activation of heterocyclic amine carcinogens and support the hypothesis that the NAT2 acetylation polymorphism is a breast cancer risk factor among postmenopausal women with high levels of heterocyclic amine exposure. PMID- 11008908 TI - Tamoxifen and mammographic breast densities. AB - The extent of breast tissue density on mammograms is one of the strongest risk factors for breast cancer. The aim of this analysis was to evaluate whether tamoxifen can affect mammographic breast density. Subjects were participants in the National Surgical Adjuvant Breast Project Breast Cancer Prevention Trial (BCPT), recruited and followed at the Breast Center of Saint-Sacrement Hospital in Quebec City, Canada. The Breast Cancer Prevention Trial is a double-blind trial in which women at high risk of breast cancer were randomized to receive either 20 mg tamoxifen per day or placebo. Mammograms were taken before treatment began and yearly thereafter. For the purpose of this analysis, Wolfe's parenchymal pattern and the percentage of the breast showing tissue densities were assessed by review of pre- and posttreatment mammograms without knowledge of treatment assignment. Among the 69 women included in this analysis, 36 received tamoxifen and 33 received placebo for an average of 3.3 and 3.5 years, respectively. Among women receiving tamoxifen, 16 of 36 (44.4%) changed to a parenchymal pattern of lower density compared with 5 of 33 (15.2%) women receiving placebo (P = 0.010). Moreover, in the tamoxifen-treated group, the difference in the percentage of the breast showing tissue densities between the pre- and posttreatment mammograms reached -9.4% on average compared with a reduction of -3.6% in the placebo group (P = 0.010). Our data show that tamoxifen can reduce high-risk mammographic features. Breast densities should be evaluated as possible early markers of the preventive effect of selective estrogen receptor modulators. PMID- 11008909 TI - Effect of tamoxifen on mammographic density. AB - There are strong data showing that increased breast cancer risk is associated with increased mammographic density. Tamoxifen has been shown to decrease the risk of invasive breast cancer and decrease breast density. We sought to demonstrate and calculate the extent of change in mammographic density in women who have taken tamoxifen for up to 2 years. We evaluated mammograms from 28 high risk women who were taking tamoxifen. Four different methods of evaluation were used: (a) two qualitative methods (Wolfe criteria and the American College of Radiology Breast Imaging and Reporting Data System criteria); (b) one semiquantitative method (mammograms were assigned one of five semiquantitative scores by visual inspection); and (c) one quantitative method (computer-aided calculation of fibroglandular area from digitized mammograms). The Wolfe criteria showed a 0.03 category decrease per year (P = 0.50). The American College of Radiology Breast Imaging and Reporting Data System criteria showed a 0.1 category decrease per year (P = 0.12). Semiquantitative criteria showed a 0.2 category decrease per year (P = 0.039). Digitized scores showed a 4.3% decrease per year (P = 0.0007). In conclusion, tamoxifen causes a decrease in mammographic density with use, an effect that is better quantitated with semiquantitative criteria or digitized images. Density change might become useful as a surrogate end point for the effect of tamoxifen and other chemopreventive measures, although our data do not predict an individual's degree of risk reduction. PMID- 11008910 TI - Effectiveness and cost-effectiveness of multiple outcalls to promote mammography among low-income women. AB - A multiple outcall approach based on the Transtheoretical Model was used to encourage mammography behavior in low-income women in the state of Colorado. Women (n = 983) were recruited in person at grocery and discount stores and were then called over the telephone to receive the multiple outcall intervention. These women were compared with 3,080 women who were recruited by telephone and randomly assigned to three study groups: health survey only (control); single outcall; or advance card + single outcall. Subsequent mammography behavior was assessed through a telephone interview conducted 6 months after initiation of the protocol. After controlling for baseline differences between groups in age, education, income, health status, and previous mammography behavior, the multiple outcall intervention was significantly related to mammography behavior among women nonadherent at baseline (odds ratio, 2.58; 95% confidence interval, 1.45 4.60). Furthermore, women who received the multiple outcall intervention had higher "stage of change" at follow-up and more positive attitudes toward mammography. Cost-effectiveness analysis indicated that although the multiple outcall intervention was more costly to deliver ($14.84 per subject compared with about $7.00 for the single outcall interventions), it cost considerably less per subject converted from nonadherent to adherent. Despite study design limitations, the multiple outcall intervention appears to be an effective method of promoting mammography among previously nonadherent women. The results suggest that a combined approach, in which nonadherent women receive multiple calls promoting screening behavior, followed by single calls at the appropriate intervals to promote repeat screening, may be a useful strategy in defined populations. PMID- 11008911 TI - A case-control study of analgesic use and ovarian cancer. AB - A recent case-control study raised the hypothesis that acetaminophen use 1 day or more per week for at least 6 months reduces the risk of epithelial ovarian cancer. We assessed analgesic use in relation to epithelial ovarian cancer risk using data from our case-control surveillance study of medication use and cancer. Patients were interviewed in hospitals in Baltimore, Boston, New York, and Philadelphia during 1976-1998. We compared 780 women with epithelial ovarian cancer to 2053 cancer controls and 2570 noncancer controls. For acetaminophen use 1 day or more per week for at least 6 months, the odds ratio estimate was 0.9 (95% confidence interval, 0.6-1.4) derived with cancer controls and 1.0 (0.6-1.5) with noncancer controls. Estimates for more frequent and longer term use were also compatible with 1.0. The odds ratios among patients with metastatic ovarian cancer were reduced but not statistically significant. The odds ratio for use of nonsteroidal anti-inflammatory drugs 4 or more days per week for at least 5 years, 0.5, was statistically significant. The present results provide only weak support for a reduction in the risk of epithelial ovarian cancer among acetaminophen users. They raise the possibility of an inverse association with long-term nonsteroidal anti-inflammatory drug use. PMID- 11008912 TI - Precision and bias of food frequency-based measures of fruit and vegetable intakes. AB - Accurate assessment of fruit and vegetable intakes is critical for cancer control research and public health surveillance. This report examines the bias and precision of two commonly used food frequency questionnaire methods to assess fruit and vegetable intakes: (a) the 5 A Day method, based on seven items; and (b) the summation method, based on adding total servings of all fruit and vegetable items on a comprehensive (100+ item) food frequency questionnaire. Data are from three studies in which 24-h dietary recalls, food records, or serum carotenoid concentrations could be used as criterion measures (n = 260, 1031, and 342). Studies differed markedly in distributions of participants' age, race/ethnicity, sex, and socioeconomic status. Mean intakes of total fruit and vegetables based on the 5 A Day method were consistently lower than those from either the summation method (3.11 versus 4.06), 24-h recalls (3.32 versus 4.07), or food records (3.11 versus 3.46; all P < 0.01), and this was due primarily to underestimates of vegetable intake. Correlations of the 5 A Day and summation measures with all criterion measures were similar and were consistently higher for fruit (range, 0.33-0.57) than for vegetables (range, 0.24-0.32). These results, which were consistent across diverse participant samples, suggest that the 5 A Day method yields both biased and imprecise measures of vegetable intake and that research to improve this measure is needed. PMID- 11008913 TI - Human papillomavirus testing for primary screening of cervical cancer precursors. AB - Our objective was to determine whether the addition of human papillomavirus (HPV) testing to screening cytology improves the detection of cervical cancer precursors. Women of ages 18-69 years underwent conventional Pap cytology and HPV DNA testing in a multicenter study in Newfoundland, Canada. Those with positive cytology and/or HPV and a random sample of those with dual negative results were referred for colposcopy. The study enrolled 2098 women. The relative sensitivity of HPV testing was significantly higher than cytology for all-grade squamous intraepithelial lesions [SILs; 73%; 95% confidence interval (CI), 62-82] and high grade SILs (HSILs; 90%; 95% CI, 74-97) but had lower relative specificity (62% for all-grade SILs and 51% for HSILs) than most cytological cutpoints. The rate of combined correct results for all-grade lesions was higher for HPV testing (68.8%) than for any cytological cutpoint (equivocal, 52.3%; LSILs, 51.6%; HSILs, 44.5%). The combination of HPV and an LSIL cutpoint had a negative predictive value of 68% (95% CI, 52-80) for all SILs and 100% (95% CI, 91-100) for HSILs, while referring for colposcopy only 12% of the women. We concluded that HPV testing in conjunction with cytology improved the screening efficacy of cytology alone and may allow for a more effective and safe primary screening program with increased screening intervals. PMID- 11008914 TI - Correlates of the spread of human papillomavirus infection. AB - Knowledge of the correlates of human papillomavirus (HPV) seropositivity is of interest for planning of preventive measures and for evaluation of possible confounding in epidemiological studies. The epidemiological determinants for seropositivity for oncogenic and benign HPV types were assessed using a serosurvey of 275 healthy Swedish women, stratified by age and lifetime number of sexual partners. Seroprevalences were compared with 17 behavioral variables obtained by interview and 3 laboratory-diagnosed microbiological exposures. In univariate analysis, history of gonorrhea and condylomatosis, human herpesvirus type 8 and herpes simplex virus 2 seropositivities, lifetime number of sexual partners, and current partner's lifetime number of sexual partners were associated with oncogenic HPV seropositivity. Noteworthy lack of correlations included smoking habits and oral contraceptive use. In multivariate analysis, only the number of lifetime sexual partners [odds ratio (OR), 8.7; 95% confidence interval (CI), 3.3-22.6] and seropositivity for benign HPV types remained significant (OR, 2.9; 95% CI, 1.6-5.3). Seropositivity for benign HPV was primarily associated with condyloma history (OR, 3.6; 95% CI, 1.2-10.8) and seropositivity for oncogenic HPV (OR, 2.9; 95% CI, 1.6-5.2). An association with sexual history lost significance in the multivariate model. In conclusion, lifetime number of sexual partners is the major determinant of acquisition of oncogenic HPV. By contrast, benign HPV infection associates more strongly with condyloma history than with sexual history per se. PMID- 11008915 TI - Helicobacter pylori and stomach cancer: a case-control study in Venezuela. AB - The role of Helicobacter pylori infection in gastric cancer was evaluated in a high-risk population in Venezuela using serological assays in a study of 302 cases and 483 neighborhood controls. To investigate the claim that assays for H. pylori should use antigens derived from local strains, four different assays derived from Venezuelan and European strains were used. Prevalence of IgG H. pylori antibodies in controls was very high, with estimates between 72 and 92%. Prevalence was similar in cases and controls. However, cases had lower antibody titers. This effect was observed only in subjects with low pepsinogen (PG) levels PGI/PGII <3.0), which suggested that extensive atrophy in cases causes a loss of H. pylori infection, with a consequent reduction in antibody titer. In addition, advanced cases (stage II or higher) had lower antibody titers than less advanced cases, which indicated that the lower antibody titers in cases may be attributable partially to a diminished immune response. All of the four assays for anti-H. pylori antibodies gave similar results. No evidence was found for the superiority of the assay based on Venezuelan strains. These results are consistent with other case-control studies in high-risk populations and highlight the difficulties of investigating H. pylori infection in retrospective studies. PMID- 11008916 TI - Parental smoking and alcohol consumption and risk of neuroblastoma. AB - Previous studies and animal evidence have suggested a relationship between parental tobacco or alcohol use and the risk of some childhood cancers, including neuroblastoma. A case-control study was conducted to investigate the relationship between parental tobacco smoking, alcohol consumption, and risk of neuroblastoma. Cases were children diagnosed with neuroblastoma over the period 1992-1994 at Children's Cancer Group and Pediatric Oncology Group institutions throughout the United States and Canada. One matched control was selected using random-digit dialing. Information on parental smoking and drinking history was obtained from 504 case and 504 control parents by telephone interview. Overall, there was no consistent pattern of association with parental smoking and alcohol consumption. For example, both maternal smoking and drinking during the period from 1 month before pregnancy through breastfeeding had adjusted odds ratios (ORs) of 1.1 [95% confidence interval (CI), 0.8-1.4]. There was no association with paternal smoking (OR, 1.2; 95% CI, 0.8-1.6) or paternal drinking 1 month before conception (OR, 1.0; 95% CI, 0.7-1.4). There was no consistent increase in risk by the amount of smoking or drinking during any time period relative to pregnancy. There was no suggestion of an increased risk when only one parent smoked. Smoking or drinking among both parents did not jointly increase the risk of neuroblastoma in their offspring. The child's age at diagnosis, stage, or MYCN oncogene amplification status did not materially alter the OR estimates. It is concluded that the results from this study do not indicate any evidence for a relationship between neuroblastoma and parental tobacco or alcohol use. PMID- 11008917 TI - Relationships among breast cancer perceived absolute risk, comparative risk, and worries. AB - When trying to predict breast cancer screening, it may be important to understand the relationships between perceived breast cancer risks and worries about getting breast cancer. This study examines the extent to which women's worries about breast cancer correlate with perceptions of both absolute (assessment of own) and comparative (self versus other) 10-year and lifetime risks. As part of a larger randomized intervention trial concerning hormone replacement therapy, 581 women participated in a telephone baseline survey to assess their perceptions of breast cancer risks and worries. Worries about getting breast cancer in the next 10 years and in one's lifetime were related positively to both absolute and comparative 10-year and lifetime risks. The magnitude of these relationships did not differ by time frame. Worry about breast cancer is a function of both how a woman views her own risk and how she compares her risk with that of other women. Some practitioners may encourage women to get screened for breast cancer by using emotional appeals, such as heightening women's worries about breast cancer by using risk information. Our data suggest that they should give careful consideration how best to combine, if at all, information about absolute and comparative risks. For example, if the motivation to screen is based on a sequential assessment of risk beginning with comparative and then absolute risk, creating communications that heighten perceived risk on both of these risk dimensions may be needed to evoke sufficient worry to initiate breast cancer screening. PMID- 11008918 TI - Consumer perception of risk associated with filters contaminated with glass fibers. AB - The filters in Eclipse, a new cigarette-like smoking article marketed by R. J. Reynolds Tobacco Company, are contaminated with glass fibers, fragments, and particles. Reported herein are the results of a study in which consumers were questioned about their opinions as to whether exposure to glass fibers in such a filter poses an added health risk beyond that from smoking and whether the manufacturer has an obligation to inform consumers about the glass contamination problem. The study queried 137 adults who were interviewed while waiting at a Division of Motor Vehicles office in Erie County, New York in 1997. All but one person expressed the view that the presence of glass fibers on the filters poses an added health risk beyond that associated with exposure to tobacco smoke alone. Nearly all expressed the position that the cigarette manufacturer has a duty to inform the public about the potential for glass exposure. PMID- 11008919 TI - Helicobacter pylori strain types and risk of gastric cancer: a case-control study. AB - The aim of this novel endoscopy clinic-based case-control study was to explore the influence of different Helicobacter pylori strain types on the risk of gastric adenocarcinoma using isolated bacterial strains, tissue samples, and sera. We included 72 cases with gastric adenocarcinoma and 324 age- and sex matched controls. Histological characterization, culture, molecular typing of H. pylori genes by PCR (cagA/vacA), conventional IgG ELISA, and immunoblotting (Western blot) for the CagA and VacA proteins were performed. With four tests combined, H. pylori infection was detected in 57 (79%) cases and 213 (66%) controls. A positive association between H. pylori infection and gastric cancer risk was found [odds ratio (OR), 2.1; 95% confidence interval, 1.1-3.9]. Type I (OR, 1.8), intermediate (OR, 2.0), and type H (OR, 0.2) strains of H. pylori presented different serum antibody levels and different levels of association with gastric cancer. Our case-control study, based on molecular characterization and serology, provides further evidence that infection by more virulent strains of H. pylori and the presence of antibodies toward the CagA protein can be used as markers for an increased risk of gastric adenocarcinoma and that the strain types of H. pylori could be used in the future to determine disease outcome. PMID- 11008920 TI - CYP1A1 I462V genetic polymorphism and lung cancer risk in a cohort of men in Shanghai, China. AB - Cytochrome P450 (CYP) CYP1A1 activates tobacco-related carcinogens. A point mutation at codon 462 in exon 7 of CYP1A1 results in a substitution of isoleucine by valine near the heme binding site. This mutation is rare in Caucasians but common in Japanese populations, in which association with increased risk of lung cancer has been reported. There are few data in other Asian populations. We investigated this I462V polymorphism using DNA from 214 incident cases of lung cancer and 669 controls in a prospective cohort study of 18,244 middle-aged and older men in Shanghai, China. The valine allele frequency was 0.138 among the control population. The I462V genotype was not appreciably associated with lung cancer risk overall. There was some suggestion that having at least one valine allele might be related to increased risk of lung cancer among smokers of <20 cigarettes/day (odds ratio, 1.72; 95% confidence interval, 0.82-3.62), particularly among those with homozygous deletion of GSTM1 (odds ratio, 2.80; 95% confidence interval, 1.07-7.33), which is involved in the detoxification of activated tobacco carcinogens. In this Chinese cohort, with CYP1A1 valine allele frequency intermediate between Japanese and Caucasian populations, the I462V polymorphism is not related to lung cancer overall, but it might play a role at lower levels of cigarette smoking among subjects with impaired carcinogen detoxification as assessed by the GSTM1-null genotype. PMID- 11008921 TI - Expression of protein mediators of type I interferon signaling in human squamous cell carcinoma of the skin. AB - IFN-based therapy has been shown to be active in the treatment of squamous cell carcinoma (SCC) of the skin and has promise for chemoprevention and treatment of several other cancers. In an effort to better understand the molecular mechanism of this activity, we have determined the expression pattern of several of the protein mediators of type I IFN signaling by immunohistochemistry in cutaneous SCC, SCC metastases, and adjacent nonmalignant epithelium from patient biopsies. All of the proteins, signal transducer and activator of transcription (STAT) 1alpha/beta, STAT2, p48, STAT3a, and STAT3beta, are expressed at varying levels in the adjacent epidermis, as well as in other epidermal and dermal cell types. For the majority of samples tested, the expression of one or more of these proteins was reduced in SCC primary tumors compared with the adjacent nonmalignant epithelial cells, as determined by manual scoring. Quantitative densitometry of several samples revealed differences that are statistically significant. Our study provides the first direct evidence for the expression of the IFN-stimulated gene factor 3 (STAT1alpha/beta, STAT2, and p48) and STAT3alpha and STAT3beta mediators of IFN-alpha/beta signaling in human skin and skin derived SCCs. These data have led to the hypothesis that the loss of IFN sensitivity may contribute to the development and progression of skin SCC. PMID- 11008923 TI - Hydrochloric attack of serpentinites: Mg2+ leaching from serpentinites. AB - This paper presents the hydrochloric attack of serpentinites. Romanian serpentinites are part of the Danubian crystalline complex. These serpentinites contain mineral serpentine (magnesium silicate) and a small amount of other minerals along with calcium, iron, aluminium, nickel, cobalt, chrome and manganese. For processing, the serpentinite was attacked by hydrochloric acid from two provinces (p.a. and residual hydrochloric acid), using different experimental conditions: acid concentrations between 15-23 per cent HCl, the temperature range of 70-90 degrees C, with stoichiometric ratio between reactants and with acid excess, the extraction time being up to 180 min. In optimum conditions, the magnesium extraction is almost total (99.5 per cent). A well balanced processing of these serpentinites looks for both the silica residue utilization and the chloride solutions, in order to separate the prevalent magnesium as well as the valuable microcomponents--nickel and cobalt. PMID- 11008922 TI - Predictors of inactivation and reasons for participant inactivation during a skin cancer chemoprevention study. AB - Maintaining good compliance is a major challenge in long-term cancer chemoprevention trials. Minimizing the number of inactive participants during a trial is an important factor in maximizing compliance. Identifying reasons for and predictors of inactivation is the first step in being able to reduce participant inactivation. In this skin cancer chemoprevention trial, the 2,297 participants were randomized to receive 25,000 IU of retinol daily or a placebo. Median follow-up time was 3.8 years. The reason for inactivation was determined for each participant who stopped taking the study capsules. Six hundred and seventy-seven (29.7%) participants became inactive during the 5-year study. There was no significant difference between the number of participants inactivating by treatment group or sex. The most common reasons for inactivation were illness of subject, spouse, or a close relative (18.6%) and experience of a clinical symptom consistent with vitamin A ingestion (17.1%). Participants in the vitamin A group (10.1%) more frequently cited symptoms coded as "not consistent with vitamin A" as the reason for inactivation compared with those in the placebo group [5.4% (P < 0.05)]. The inactivation rate was highest in the first month of the trial and declined thereafter. A low education level (hazard ratio, 1.59) and unmarried status (hazard ratio, 1.29) were the only significant predictors of inactivation. These findings may be useful in developing targeted strategies to decrease inactivation and thereby increase compliance in future chemoprevention trials. However, these findings need to be confirmed because published research in this area is very limited. PMID- 11008924 TI - Association effects of vitamin B6 and various magnesium salts on a pharmacological model: the human amniotic membrane. AB - The effects of the association of vitamin B6 and Mg salts: aspartate, citrate, lactate, pidolate, sulfate, were studied on the ionic transfer through a membraneous pharmacological model: the isolated human amniotic membrane. The ionic transfer was evaluated by measure of the total conductance in the maternal to fetal way (GtM) and in the fetal to maternal way (GtF), of the ionic fluxes (F1 on the maternal side, F2 on the fetal side) and of the ratio F1/F2. The results were explained in terms of monophasic (decrease: screening or increase: binding interactions with the polar surface moities) or biphasic (decrease then increase) action. The results indicated: Mg aspartate decreased GtF, F1, F2 whatever concentration and had a concentration-dependent effect on GtM, F1/F2. The addition of vitamin B6 induced a new concentration-dependent effect (biphasic action: decrease then increase) on GtF, F1 and also modified F2, F1/F2. Mg citrate decreased GtM, GtF, F2 whatever the concentration and had a concentration dependent effect on F1, F1/F2. The addition of vitamin B6 induced a new biphasic effect on GtM and GtF. Mg lactate decreased GtM, F1, F2, F1/F2 whatever the concentration but had a concentration-dependent effect on GtF. The addition of vitamin B6 induced a new biphasic effect on GtM, F1, F2, F1/F2. Mg pidolate had no effect on GtM, GtF, F2, F1/F2 and decreased F1. The addition of vitamin B6 did not induce variation. Mg sulfate had no effect on GtM, increased GtF and decreased F1, F2, F1/F2. The addition of vitamin B6 induced a new concentration dependent effect on F2. The association between vitamin B6 and Mg salts implicated a new action on components of the amniotic ionic transfer characterized by a biphasic action (decrease then increase concentration dependent effect). This effect was dependent on the anion associated with magnesium. The magnesium salts may be classified with regard to the beneficial effect due to the association with vitamin B6 in the following decreasing order: aspartate and lactate, citrate, pidolate, sulfate. PMID- 11008925 TI - Importance of decreased intracellular phosphate and magnesium concentrations and reduced ATPase activities in spontaneously hypertensive rats. AB - A decrease in total magnesium content is not a direct proof of a decreased magnesium ion concentration. It could reflect a phosphate alteration or an ATP metabolism disorder. Plasma phosphate levels are lower in spontaneously hypertensive rats (SHRs) than in Wistar-Kyoto (WKY) rats, and defects in membrane regulation or mitochondrial ATP synthase occur. Only sparse data exist concerning cellular magnesium and phosphate concentrations in hypertensive cells. In aortic smooth muscle cells from 10 SHRs of the Munster strain and 10 age-matched normotensive WKY rats, the intracellular phosphate and magnesium content was measured by electron probe X-ray microanalysis (Camscan CS 24 apparatus, Cambridge, U.K.). The Mg++ content was 0.90+/-0.15 g/kg dry weight in SHRs versus 1.15+/-0.10 g/kg dry weight in WKY rats (p<0.05). Vascular smooth muscle phosphate content was 23.6+/-0.79 g/kg dry weight in WKY rats versus 15.81+/-1.22 g/kg dryweight in SHRs (p<0.01). In seven animals, erythrocytic ATP content was 180.2+/-102 in SHRs vs. 432+/-72 micromol/L cells in WKY rats (p< 0.01). The Na+/K+-ATPase activity was significantly decreased in hypertensive animals as compared to controls (6.49+/-2.3 vs. 12.64+/-2.9 nmol inorganic phosphate/mg protein/min (p< 0.01)). Aortic smooth muscle cells from SHRs are characterized by markedly lowered cellular phosphate and magnesium concentrations and an altered ATP metabolism, possibly due to a membrane defect or a magnesium deficit in hypertensive cells. PMID- 11008926 TI - Disturbances of magnesium concentrations in various brain areas in Alzheimer's disease. AB - Our first project aimed at the determination of the average values of Mg in normal human brain (20 individuals, mean age 70 years). Ten brain parts were selected from both hemispheres. Determinations were carried out by inductively coupled plasma atomic emission spectrometry, inductively coupled plasma mass spectrometry and instrumental neutron activation analysis methods. In order to investigate the precision and the accuracy of the methods five biological standard reference materials were analyzed. The present results show a non homogeneous distribution of Mg in normal human brain. Regions corresponding to each other in both hemispheres show almost identical concentration. The second goal was to study the possible Mg concentration changes in Alzheimer's disease (nine patients, ten brain parts). Mg values are found to be significantly decreased in brain regions of diseased patients compared to the controls. PMID- 11008927 TI - Are low magnesium levels in type 1 diabetes associated with electromyographical signs of polyneuropathy? AB - OBJECTIVE: Our aim was to study the relationship between the magnesium status in type 1 diabetic patients and disturbances in nerve conduction velocity. Furthermore we wanted to investigate whether repletion of magnesium depletion could improve the decreased nerve conduction velocity measurements. In a cross sectional study, 154 type 1 diabetic patients were screened for their erythrocyte magnesium content and an electrophysiological measurement of the peripheral nervous system was carried out. In a subsequent intervention study, out of this screened population, 23 type 1 patients, with disturbed nerve conduction velocity measurements and low erythrocyte magnesium levels [< 2.3 mmol/L) were given oral magnesium supplements, during 1 year. Twenty type 1 patients with identical characteristics served as controls. In the cross-sectional study disturbed nerve conduction velocities were found in the older patients, in patients with a longer duration of diabetes and a worse metabolic control. EMG polyneuropathy signs were significantly more frequent in diabetic patients with low erythrocyte Mg. The intervention study demonstrated that under unchanged metabolic control, supplementation with magnesium could improve nerve conduction, especially in younger patients with a shorter duration of diabetes. Erythrocyte Mg was lower in type 1 diabetic patients with polyneuropathy. Mg supplementation increasing Mg RBC might (possibly?) improve nerve conduction measured by electromyography at least in younger patients with a short duration of diabetes and presenting early signs of the neurological complication. PMID- 11008928 TI - A review of the status of magnesium and related minerals in the sudden infant death syndrome (SIDS). AB - Sudden infant death syndrome (SIDS) is the sudden death of an infant under 1 year of age that remains unexplained after a thorough case investigation, including performance of a complete autopsy. Despite recent reductions in the SIDS rate attributed to placing the baby to sleep on his back, SIDS remains the most common cause of infant mortality between 1 month and 1 year of age in developed countries. This review concerns the current state of a hypothesis (1972) that magnesium deficiency, which causes sudden death in young mammals, is the major cause of SIDS. Numerous investigators have compared the concentration of magnesium, and other minerals from soft tissues and bone in SIDS with non-SIDS controls (normal infants who died suddenly of known cause such as trauma). Because of rapid shifts of minerals during early development, SIDS and control infants must be matched for gestational and postnatal age. About 95 per cent of SIDS occurs between 1-6 months of life, with peak incidence at 2-4 months, when vitreous magnesium is high. There is little change in magnesium in the relatively inert vitreous despite extremes in dietary magnesium. All values fall within a small range, with scatter. Magnesium rapidly increases in foetal cartilage with ossification of the bone. Early in magnesium deficiency, liver magnesium may be increased or unchanged, but it does not increase during magnesium excess. Lead accumulation is increased in magnesium deficiency. Among infants with high environmental exposure to lead, those who die of SIDS rather than non-SIDS infants have the greater lead burden. Soft water with low magnesium and calcium and with high concentration of sodium have been linked to higher SIDS rates, which have been attributed to low magnesium. It is concluded that the SIDS hypothesis has neither been proved nor disproved. PMID- 11008929 TI - The Windscale accident--some memories and reflections. PMID- 11008930 TI - Epidemiological studies of the Sellafield workforce. PMID- 11008931 TI - Mortality and cancer registration experience of the Sellafield employees known to have been involved in the 1957 Windscale accident. AB - The mortality and cancer morbidity experience of the 470 male Sellafield employees known to be involved in the 1957 Windscale accident is reported. All these employees are known to have been involved in dealing with the fire itself, or in the clean-up operation afterwards. The size of the study population is small, leading to predicted low power to reveal any effects, but the cohort is of interest because of the involvement of the workers in the accident. For 1957-97, using rates for England and Wales to calculate the expected numbers, the all causes standardised mortality ratio (SMR) is 100 (observed = 258, expected = 258.80), and the all malignant neoplasms SMR is 79 (observed = 58, expected = 73.12) which is not significantly different from 100. For 1971-91, the all malignant neoplasms standardised registration ratio (SRR) of 85 (observed = 59, expected = 69.23) is not significantly different from 100. Significant excesses of deaths from diseases of the circulatory system (SMR = 121) and from ischaemic heart disease (SMR = 128), and a significant deficit of deaths from cancer of the genito-urinary organs (SMR = 31), were found. There were no significant differences in mortality rates between workers who had received high recorded external doses during the fire and those who had received low doses, though the power of this comparison was low. Comparison of the mortality rates of workers directly involved in the accident with workers in post, but not so involved, showed no significant differences. This study has been unable to detect any effect of the 1957 fire upon the mortality and cancer morbidity experience of those workers involved in it. PMID- 11008932 TI - The assessment of organ doses from plutonium for an epidemiological study of the Sellafield workforce. AB - An epidemiological study of mortality and cancer rates in plutonium workers at the Sellafield nuclear installation has been carried out by researchers at the London School of Hygiene and Tropical Medicine. The study required the assessment of organ-specific doses from plutonium for more than 5000 workers over a period of 40 years. This was a major undertaking as it involved the reconstruction of annual received doses from the results of some 223,000 urine samples that had been provided by the workers in the study. This paper outlines the techniques and strategies adopted in order to generate best estimates of dose from the available data, and presents summaries of these doses. PMID- 11008933 TI - An optical method for three-dimensional dosimetry. AB - Accurate determination of the spatial distribution of the absorbed dose of ionising radiation plays an important role in radiotherapy, industrial radiation processing and many other applications. Computer calculations have frequently been used to estimate three-dimensional (3D) dose distributions in complex geometries and it becomes important to validate these by accurate 3D measurements. For this purpose we have been investigating the use of gelatin gels loaded with a modified Fricke solution which are pale orange in appearance and which, upon irradiation, become increasingly purple when viewed in normal light. This ferrous sulphate xylenol orange in gelatin gel (FXG) system displays very good properties, such as sensitivity, linearity and dynamic range, that make it suitable for 3D dosimetry applications. A high-speed optical tomography readout technique has been developed enabling two-dimensional projections of optical absorption data to be recorded rapidly. From these data the 3D absorbed dose distribution can quickly be derived with minimal degradation due to ion diffusion. PMID- 11008934 TI - Prominent artificial radionuclide activity in the environment of coastal Karnataka on the southwest coast of India. AB - Studies on radiation level and radionuclide distribution in the environment of coastal Karnataka were undertaken to provide baseline data for the future assessment of the impact of the nuclear and thermal power stations that are being set up in the region and to understand the behaviour of radionuclides in the environment. As part of the programme the concentrations of two important artificial radionuclides, namely 90Sr and 137Cs, have been measured in a number of environmental samples. The concentration of 90Sr is very low in most of the samples. Among the samples analysed for the concentration of 137Cs, soil samples showed elevated levels of activity in some sampling stations. Among the vegetables, brinjal (Solanum melongena. L) showed considerable activity. The internal dose due to intake of 90Sr through diet was 0.42 microSv year(-1) for the vegetarian population and 0.32 microSv year(-1) for the non-vegetarian population. The internal dose due to dietary intake of 137Cs was found to be 0.34 microSv year(-1) and 0.26 microSv year(-1) respectively for the vegetarian and non-vegetarian population. The results are discussed in the light of the literature values reported for other environs of India and abroad and appropriate inferences are drawn. PMID- 11008935 TI - Contaminated land liability issues: legal perspective. AB - Until recently, companies that contaminated land faced little prospect of being prosecuted or having claims made against them for any personal injuries or property damage arising from the contamination. Most companies would have considered it to be inconceivable that they could be required to clean up contamination resulting from an historic disposal, particularly if the disposal had been neither negligent nor unlawful. Current environmental law differs significantly, however, from the law which existed even 10 years ago. This paper examines liabilities arising from contaminated land. First, it discusses the new contaminated land regime and a parallel regime to remediate sites that are contaminated with radioactive substances. Second, it discusses remediation liabilities for water pollution. Such liabilities may well involve the remediation of contaminated land when groundwater is polluted. Finally, the paper examines liabilities for property damage and personal injuries arising from contamination by radioactive substances. PMID- 11008936 TI - Application of regulations to cosmetic lasers in private practice. AB - Recent years have seen a growth in the number of non-NHS premises, for example beauty salons, offering cosmetic laser treatments such as laser hair removal. By law, such practices are subject to registration and monitoring by Health Authorities. However, a national consensus on appropriate standards is urgently required, particularly on interpretation of the legal requirements for medical direction and operator training, in order to ensure consistency of standards in private practice with lasers across the country. This paper outlines the difficulties experienced in the inspection and registration process, and makes recommendations on appropriate interim standards, pending formal guidance from professional bodies and the promulgation of revised regulations. Until such guidance is in place, it may be wise for Health Authorities to include within their inspection protocols a requirement that applicants for laser registrations produce a statement as to the exact form which medical direction of laser treatment will take, to enable the Health Authority to consider and take advice on its suitability. Similarly a requirement that laser operators are state registered health professionals will ensure that these potentially hazardous devices are only used by properly qualified operators, whilst professional bodies work towards a consensus on minimum training standards. PMID- 11008937 TI - Stillbirths and male irradiation. PMID- 11008938 TI - On collective dose. PMID- 11008939 TI - The 'controllable dose' debate: results of the IRPA consultation exercise. PMID- 11008940 TI - Study of childhood cancer and paternal preconceptional irradiation at USA nuclear facilities. PMID- 11008941 TI - Japanese atomic bomb survivors. PMID- 11008942 TI - SRP meeting: Surface and airborne contamination. AWE Aldermaston, 26-27 June 2000. PMID- 11008943 TI - Conference on medical physics and its applications. Zarka-Jordan, 24-26 April 2000. PMID- 11008944 TI - Commentary: the controversy between the IAEA Code of Practice and the TG-51 protocol. PMID- 11008945 TI - Power frequency electromagnetic fields and health. Where's the evidence? AB - Twenty years ago concerns were raised that exposure to power frequency (or extremely low frequency (ELF)) electromagnetic fields (EMFs) may be associated with an increased risk of cancer or other health hazards. Subsequently no associations have been shown between laboratory magnetic field exposures and carcinogenesis in either animal or cellular models. Indeed, studies have demonstrated that magnetic fields are not associated with cancer. However, the puzzle remains that the results of some epidemiological studies may be interpreted as suggesting that living close to high-voltage transmission (HVT) lines appears to increase slightly the risk of childhood leukaemia. Alternatively, these results could result from small biases and errors in individual studies, which might not necessarily be the same in each study. The nature of the epidemiological studies (power-line, wire code, magnetic field or appliance based) appears to determine whether and how the EMFs associated with HVT lines might be a risk factor. It is possible that a simple association with either magnetic or electric field exposure may not be the whole answer, and an alternative mechanism is always a possibility. Although the interpretation of the available evidence by most expert bodies has led them to conclude that exposure to power frequency electric and magnetic fields is not a human health hazard, a working group under the auspices of the US National Institute of Environmental Health Sciences (NIEHS) concluded that there was a possible low risk associated with certain exposures to ELF magnetic fields. NIEHS itself interpreted the finding as insufficient to warrant aggressive regulatory concern but stated that, because virtually everyone is routinely exposed to ELF EMFs, passive regulatory action is warranted, such as a continued emphasis on educating both the public and the regulated community on means aimed at reducing exposures. These analyses, conclusions and advice are not contradicted by subsequent studies, and therefore the conclusion of the World Health Organisation that further research is needed seems valid. PMID- 11008946 TI - A comparison between calculated and experimental kQ photon beam quality correction factors. AB - To validate the calculated values of kQ for high-energy photon beams given in the International Code of Practice for radiotherapy dosimetry based on water-absorbed dose standards, a comparison with experimental values derived in standards laboratories and in clinical beams has been made. The study includes a compilation of experimental values for ionization chambers of the type NE2561/2611, NE2571, PTW30001 and PR06. The energy dependence of the G(Fe3+) ratio of high-energy x-rays to 60Co gamma-rays by Klassen et al is taken into account for all the Fricke-derived values. For three of the chamber types analysed, the comparison shows that the calculated values are a very good estimate of the average values of kQ in the entire range of photon beam qualities available for clinical use. For the NE2571 chamber type a difference which increases with energy between calculated and experimental kQ factors has been observed; however, the largest difference with a fit describing the entire set of experimental data is always smaller than 0.4%. It is concluded that if the recommendation of the Code of Practice for an individual calibration of the user's chamber at a range of photon beam qualities is not available, the use of calculated kQ factors will yield absorbed dose to water determinations accurate within the uncertainty limits of the majority of experimental data available. The good agreement between calculated and measured values, obtained for practically all the experimental data using TPR(20,10) as photon beam quality specifier, is not satisfied in some cases for two high-energy soft beams used at the Canadian NRC. There appears to be no justification for a change to a different photon beam quality specifier solely on the grounds that such a limited set of data is not described by the same distributions as the rest of the experimental data. PMID- 11008947 TI - First human transvenous coronary angiography at the European Synchrotron Radiation Facility. AB - The first operation of the European Synchrotron Radiation Facility (ESRF) medical beamline is reported in this paper. The goal of the angiography project is to develop a reduced risk imaging technique, which can be used to follow up patients after coronary intervention. After the intravenous injection of a contrast agent (iodine) two images are produced with monochromatic beams, bracketing the iodine K-edge. The logarithmic subtraction of the two measurements results in an iodine enhanced image, which can be precisely quantified. A research protocol has been designed to evaluate the performances of this method in comparison with the conventional technique. Patients included in the protocol have previously undergone angioplasty. If a re-stenosis is suspected, the patient is imaged both at the ESRF and at the hospital with the conventional technique, within the next few days. This paper reports the results obtained with the first patients. To date, eight patients have been imaged and excellent image quality was obtained. PMID- 11008948 TI - IPEM guidelines on dosimeter systems for use as transfer instruments between the UK primary dosimetry standards laboratory (NPL) and radiotherapy centres. AB - United Kingdom dosimetry codes of practice have traditionally specified one electrometer for use as a secondary standard, namely the Nuclear Enterprises (NE) 2560 NPL secondary standard therapy level exposure meter. The NE2560 will become obsolete in the foreseeable future. This report provides guidelines to assist physicists following the United Kingdom dosimetry codes of practice in the selection of an electrometer to replace the NE2560 when necessary. Using an internationally accepted standard (BS EN 60731:1997) as a basis, estimated error analyses demonstrate that the uncertainty (one standard deviation) in a charge measurement associated with the NE2560 alone is approximately 0.3% under specified conditions. Following a review of manufacturers' literature, it is considered that modern electrometers should be capable of equalling this performance. Additional constructural and operational requirements not specified in the international standard but considered essential in a modern electrometer to be used as a secondary standard are presented. PMID- 11008949 TI - Comparison of conformal radiation therapy techniques within the dynamic radiotherapy project 'Dynarad'. AB - The objective of the dynamic radiotherapy project 'Dynarad' within the European Community has been to compare and grade treatment techniques that are currently applied or being developed at the participating institutions. Cervical cancer was selected as the tumour site on the grounds that the involved organs at risk, mainly the rectum and the bladder, are very close to the tumour and partly located inside the internal target volume. In this work, a solid phantom simulating the pelvic anatomy was used by institutions in Belgium, France, Greece, Holland, Italy, Sweden and the United Kingdom. The results were evaluated using both biological and physical criteria. The main purpose of this parallel evaluation is to test the value of biological and physical evaluations in comparing treatment techniques. It is demonstrated that the biological objective functions allow a much higher conformality and a more clinically relevant scoring of the outcome. Often external beam treatment techniques have to be combined with intracavitary therapy to give clinically acceptable results. However, recent developments can reduce or even eliminate this need by delivering more conformal dose distributions using intensity modulated external dose delivery. In these cases the reliability of the patient set-up procedure becomes critical for the effectiveness of the treatment. PMID- 11008950 TI - Monte Carlo verification of IMRT dose distributions from a commercial treatment planning optimization system. AB - The purpose of this work was to use Monte Carlo simulations to verify the accuracy of the dose distributions from a commercial treatment planning optimization system (Corvus, Nomos Corp., Sewickley, PA) for intensity-modulated radiotherapy (IMRT). A Monte Carlo treatment planning system has been implemented clinically to improve and verify the accuracy of radiotherapy dose calculations. Further modifications to the system were made to compute the dose in a patient for multiple fixed-gantry IMRT fields. The dose distributions in the experimental phantoms and in the patients were calculated and used to verify the optimized treatment plans generated by the Corvus system. The Monte Carlo calculated IMRT dose distributions agreed with the measurements to within 2% of the maximum dose for all the beam energies and field sizes for both the homogeneous and heterogeneous phantoms. The dose distributions predicted by the Corvus system, which employs a finite-size pencil beam (FSPB) algorithm, agreed with the Monte Carlo simulations and measurements to within 4% in a cylindrical water phantom with various hypothetical target shapes. Discrepancies of more than 5% (relative to the prescribed target dose) in the target region and over 20% in the critical structures were found in some IMRT patient calculations. The FSPB algorithm as implemented in the Corvus system is adequate for homogeneous phantoms (such as prostate) but may result in significant under or over-estimation of the dose in some cases involving heterogeneities such as the air-tissue, lung-tissue and tissue-bone interfaces. PMID- 11008951 TI - Monte Carlo simulation of dose distributions from a synchrotron-produced microplanar beam array using the EGS4 code system. AB - Microbeam therapy is established as a general concept for brain tumour treatment. A synchrotron based x-ray source was chosen for experimental research into microbeam therapy, and therefore new simulations were essential for investigating the therapy parameters with a proper description of the synchrotron radiation characteristics. To design therapy parameters for tumour treatments, the newly upgraded LSCAT (Low energy SCATtering) package of the EGS4 Monte Carlo simulation code was adapted to develop an accurate self-written user code for calculating microbeam radiation dose profiles with a precision of 1 microm. LSCAT is highly suited to this purpose due to its ability to simulate low-energy x-ray transport with detailed photon interactions (including bound electron incoherent scattering functions, and linear polarized coherent scattering). The properties of the synchrotron x-ray microbeam, including its polarization, source spectrum and beam penumbra, were simulated by the new user codes. Two concentric spheres, an inner sphere, defined as a brain, and a surrounding sphere, defined as a skull, represented the phantom. The microbeam simulation was tested using a 3 x 3 cm array beam for small treatment areas and a 6 x 6 cm array for larger ones, with different therapy parameters, such as beam width and spacing. The results showed that the microbeam array retained an adequate peak-to-valley ratio, of five times at least, at tissue depths suitable for radiation therapy. Dose measurements taken at 1 microm resolution with an 'edge-on' MOSFET validated the basics of the user code for microplanar radiation therapy. PMID- 11008952 TI - Wall correction factors for calibration of plane-parallel ionization chambers with high-energy photon beams. AB - Most dosimetry protocols recommend that calibration of plane-parallel ionization chambers be performed in an electron beam of sufficiently high energy by comparison with cylindrical chambers. For various plane-parallel chambers, the 1997 IAEA TRS-381 protocol includes an overall perturbation factor pQ for electron beams, a wall correction factor p(wall) for a 60Co beam and the product of two wall corrections k(att)k(m) for 60Co in-air calibration. The recommended values of p(wall) for plane-parallel chambers, however, are limited to certain phantom materials and a 60Co beam, and are not given for other phantom materials and x-ray beams. In this work, the p(wall) values of the commercially available NACP, PTW/Markus and PTW/Roos plane-parallel chambers in a solid water phantom have been determined with 60Co and 4 and 10 MV photon beams. The k(att)k(m) values for the NACP and PTW/Markus chambers have also been obtained. The wall correction factors p(wall) and k(att)k(m) have been determined by intercomparison with a calibrated Farmer chamber. The average value of p(wall) for these plane parallel chambers was 1.005 +/- 0.1% (1 SD) for 60Co beams and 1.007 +/- 0.2% (1 SD) for both 4 MV and 10 MV photons. The k(att)k(m) values for the NACP and PTW/Markus chambers were about 1.5% lower than other published data. PMID- 11008953 TI - The value of the PinPoint ion chamber for characterization of small field segments used in intensity-modulated radiotherapy. AB - Volume averaging and lack of electronic equilibrium complicate accurate dosimetry of small photon fields. In this paper the performance of the PinPoint ion chamber for characterizing small fields used in intensity-modulated radiotherapy (IMRT) was investigated and the results were compared with those obtained using the Markus ion chamber and a diamond detector. Sharp beam penumbras were measured for a 5 x 5 cm field defined using a cerrobend block mounted on the accelerator head. In addition, output factors were measured for a 6 MV photon beam and a variety of small rectangular fields collimated widthwise using the multileaf collimator (MLC) in combination with the back-up jaws. From this study, a reference field of 5 x 5 cm and a measuring depth of 5 cm are recommended. This is related to the over-response of the PinPoint chamber to low-energy Compton scattered photons, an effect that was investigated rigorously and turned out to limit the scope of this ionization chamber. However, taking into account some limitations, the PinPoint chamber is an excellent detector for output measurements in small fields down to 2 cm. In profile measurements the chamber causes a broadening of the measured penumbras but its spatial resolution is superior to that of the Markus chamber. PMID- 11008954 TI - Theoretical considerations to the verification of dynamic multileaf collimated fields with a SLIC-type portal image detector. AB - The verification possibilities of dynamically collimated treatment beams with a scanning liquid ionization chamber electronic portal image device (SLIC-EPID) are investigated. The ion concentration in the liquid of a SLIC-EPID and therefore the read-out signal is determined by two parameters of a differential equation describing the creation and recombination of the ions. Due to the form of this equation, the portal image detector describes a nonlinear dynamic system with memory. In this work, the parameters of the differential equation were experimentally determined for the particular chamber in use and for an incident open 6 MV photon beam. The mathematical description of the ion concentration was then used to predict portal images of intensity-modulated photon beams produced by a dynamic delivery technique, the sliding window approach. Due to the nature of the differential equation, a mathematical condition for 'reliable leaf motion verification' in the sliding window technique can be formulated. It is shown that the time constants for both formation and decay of the equilibrium concentration in the chamber is in the order of seconds. In order to guarantee reliable leaf motion verification, these time constants impose a constraint on the rapidity of the image-read out for a given maximum leaf speed. For a leaf speed of 2 cm s( 1), a minimum image acquisition frequency of about 2 Hz is required. Current SLIC EPID systems are usually too slow since they need about a second to acquire a portal image. However, if the condition is fulfilled, the memory property of the system can be used to reconstruct the leaf motion. It is shown that a simple edge detecting algorithm can be employed to determine the leaf positions. The method is also very robust against image noise. PMID- 11008955 TI - Selection and determination of beam weights based on genetic algorithms for conformal radiotherapy treatment planning. AB - A genetic algorithm has been used to optimize the selection of beam weights for external beam three-dimensional conformal radiotherapy treatment planning. A fitness function is defined, which includes a difference function to achieve a least-square fit to doses at preselected points in a planning target volume, and a penalty item to constrain the maximum allowable doses delivered to critical organs. Adjustment between the dose uniformity within the target volume and the dose constraint to the critical structures can be achieved by varying the beam weight variables in the fitness function. A floating-point encoding schema and several operators, like uniform crossover, arithmetical crossover, geometrical crossover, Gaussian mutation and uniform mutation, have been used to evolve the population. Three different cases were used to verify the correctness of the algorithm and quality assessment based on dose-volume histograms and three dimensional dose distributions were given. The results indicate that the genetic algorithm presented here has considerable potential. PMID- 11008956 TI - Polymer gel dosimetry using x-ray computed tomography: a feasibility study. AB - A new three-dimensional dosimetry technique using x-ray computed tomography (CT) to analyse polymer gels is proposed. The CT imaging is sensitive to radiation induced density changes that occur within irradiated polyacrylamide gel (PAG). In this preliminary study, a CT imaging protocol is developed to optimize CT images of PAG; the response of PAG CT number to dose (N(CT)-dose response) and the reproducibility of the response are investigated, and the use of CT to analyse PAG is compared with MRI. Experiments were conducted using two 1.5 l cylindrical PAG phantoms (3% acrylamide, 3% bis and 5% gelatin by weight), one irradiated with four intersecting 10 MV photon beams and the other with 10 sets of 6 MV parallel opposed circular radiosurgery fields. The final imaging protocol involves using optimum CT parameters (120 kVp and 200 mAs for our GE HiSpeed CT/i scanner), image averaging and background subtraction. The N(CT)-dose response is reproducible, linear up to 800-1000 cGy and is relatively insensitive to the gel temperature during imaging. The dose resolution is approximately 50 cGy for an image thickness of 10 mm. Despite the low dose resolution, preliminary results indicate that this CT technique provides accurate localization of high dose gradients such as those observed in stereotactic radiosurgery. Thus, given the availability and speed of CT scanners, the technique has the potential to be a valuable and practical 3D dose verification tool in radiation therapy. PMID- 11008957 TI - General characteristics of the use of silicon diode detectors for clinical dosimetry in proton beams. AB - The properties of silicon diode detectors, used for dosimetry in clinical proton beams, were investigated with special regard to the measurement of relative dose distributions in water. Different types of silicon diode detector were studied, and the resulting distributions of detector signal versus depth in the water phantom were compared with the corresponding distributions obtained with a plane parallel NACP ionization chamber. The measurements were performed in a proton beam with an initial energy of 173 MeV. It is shown that the Hi-p silicon detector gives a signal which is proportional to the ionization density in the silicon crystal in all parts of the Bragg curve, and for all levels of accumulated dose to the detector. This is in contrast to detectors based on n type silicon, or on low resistivity p-type silicon. After pre-irradiation, these latter detectors show a stopping-power dependent recombination, yielding an increase in the detector signal per unit dose with increasing LET. This effect leads to an over-response in the Bragg peak, which increases gradually with the accumulated detector dose. Using the Hi-p silicon diode detector, the depth ionization distribution was found to be equal to the distribution obtained with the plane-parallel NACP ionization chamber at all pre-irradiation levels, within the experimental accuracy. This implies that the quotient between the ionization in the detector and the absorbed dose to the surrounding water is equal for these detectors. PMID- 11008958 TI - The development of a phantom to determine foetal organ doses from 131I in the foetal thyroid. AB - Iodine can accumulate in the foetal thyroid from the twelfth week of gestation onwards. If the iodine taken up by the foetal thyroid is in the form of 131I then the thyroid and its proximal tissues and organs will be irradiated. Several mathematical models exist in the literature on foetal/maternal iodine kinetics. However, very few studies have been performed where the foetal thyroid had been physically modelled thus allowing the determination of foetal organ dosimetry from 131I in the foetal thyroid. Here, the development of such a physical model or phantom is described and dosimetry results obtained from the phantom are discussed. The phantom is of Perspex construction, the dimensions of which are sufficient to incorporate models of the foetus at 16, 24 and 36 weeks' gestational age. The dosimetry of two organs is presented, that of the brain and the thymus. The results show that the measured absorbed dose is comparable with that calculated using modified MIRD dosimetry and traditional methods. The results also show that the dose to the thymus is greater than that of the brain by a factor of almost 30 for 16 weeks' gestational age. PMID- 11008959 TI - A comparison of quantitative measures of arterial tortuosity using sine wave simulations and 3D wire models. AB - This paper discusses the quantitative measurement of arterial tortuosity. The measures described are a tortuosity coefficient, TC, based on the second derivative of the vessel midline and the distance factor, DF, defined as L/D - 1, where L is the vessel length and D is the straight line distance between the vessel end points. A tortuosity scale derived from sine wave simulations was used to determine the ability of these two measures to quantify tortuosity. The tortuosity coefficient was demonstrated to be an appropriate measure whilst the distance factor was found to be a measure of vessel elongation rather than tortuosity. A tomographic method of deriving three-dimensional measures of TC and DF is described using a wire model. The x, y coordinates along the length of the wire were obtained from CT slices and the three-dimensional values calculated as the geometric mean of the TC and DF values measured in the x and y directions. Rotation of the wire through 90 degrees demonstrated that the three-dimensional derivations of TC and DF were independent of the orientation of the wire. PMID- 11008960 TI - Physical characteristics of the ECAT EXACT3D positron tomograph. AB - The 'EXACT3D' positron tomograph, which is now in routine clinical research use, was developed with the aim of achieving unprecedented sensitivity, high spatial and temporal resolution and simplicity of design using proven detector technology. It consists of six rings of standard detector blocks (CTI/Siemens EXACT HR+) with 4.39 mm x 4.05 mm x 30 mm elements, giving an axial field of view (FOV) of 23.4 cm. This extended FOV and the absence of interplane septa and retractable transmission rod sources has allowed greatly simplified gantry and detector cassette design. Operation in exclusive 3D mode requires an alternative to the conventional coincidence method for transmission scanning, and a single photon approach using a hydraulically driven 137Cs point source has been implemented. The tomograph has no other moving parts. A single time frame of data without any compression is very large (> 300 Mbyte) and two approaches are employed to overcome this difficulty: (a) adjacent sinograms can be summed automatically into different combinations and (b) listmode (event-by-event) acquisition has been instituted, which is both storage efficient (particularly for acquisition of sparse data sets) and maximizes temporal resolution. The high speed I/O and computing hardware can maintain a sustained acquisition rate of about 4 million coincidence events per second. A disadvantage of the large axial FOV in 3D is the increased sensitivity to activity outside the coincidence FOV. However, this can be minimized by additional side shielding. The mean spatial resolution is 4.8 +/- 0.2 mm FWHM (transaxial, 1 cm off-axis) and 5.6 +/- 0.5 mm (axial, on-axis). Its absolute efficiency is 5.8% for a line source in air (just spanning the axial FOV) and 10% for a central point source (with thresholds of 350-650 keV). For a uniform 20 cm diameter cylinder, the efficiency is 69 kcps kBq(-1) ml(-1) (after subtraction of a scatter fraction of 42%). Sensitivity relative to the EXACT HR+ (with four rings of blocks) is 2.5 (3D) and 12 (2D) times respectively. The rate of random events in blood flow studies in the brain and body, using 15O-labelled water, can be controlled by limiting the administered dose and inserting additional side shielding. PMID- 11008961 TI - Factor analysis with a priori knowledge--application in dynamic cardiac SPECT. AB - Two factor analysis of dynamic structures (FADS) methods for the extraction of time-activity curves (TACs) from cardiac dynamic SPECT data sequences were investigated. One method was based on a least squares (LS) approach which was subject to positivity constraints. The other method was the well known apex seeking (AS) method. A post-processing step utilizing a priori information was employed to correct for the non-uniqueness of the FADS solution. These methods were used to extract 99mTc-teboroxime TACs from computer simulations and from experimental canine and patient studies. In computer simulations, the LS and AS methods, which are completely different algorithms, yielded very similar and accurate results after application of the correction for non-uniqueness. FADS obtained blood curves correlated well with curves derived from region of interest (ROI) measurements in the experimental studies. The results indicate that the factor analysis techniques can be used for semi-automatic estimation of activity curves derived from cardiac dynamic SPECT images, and that they can be used for separation of physiologically different regions in dynamic cardiac SPECT studies. PMID- 11008962 TI - A filtering approach to image reconstruction in 3D SPECT. AB - We present a new approach to three-dimensional (3D) image reconstruction using analytical inversion of the exponential divergent beam transform, which can serve as a mathematical model for cone-beam 3D SPECT imaging. We apply a circular cone beam scan and assume constant attenuation inside a convex area with a known boundary, which is satisfactory in brain imaging. The reconstruction problem is reduced to an image restoration problem characterized by a shift-variant point spread function which is given analytically. The method requires two computation steps: backprojection and filtering. The modulation transfer function (MTF) of the filter is derived by means of an original methodology using the 2D Laplace transform. The filter is implemented in the frequency domain and requires 2D Fourier transform of transverse slices. In order to obtain a shift-invariant cone beam projection-backprojection operator we resort to an approximation, assuming that the collimator has a relatively large focal length. Nevertheless, numerical experiments demonstrate surprisingly good results for detectors with relatively short focal lengths. The use of a wavelet-based filtering algorithm greatly improves the stability to Poisson noise. PMID- 11008963 TI - Dose efficiency and low-contrast detectability of an amorphous silicon x-ray detector for digital radiography. AB - The effect of dose reduction on low-contrast detectability is investigated theoretically and experimentally for a production grade amorphous silicon (a-Si) x-ray detector and compared with a standard thoracic screen-film combination. A non-prewhitening matched filter observer model modified to include a spatial response function and internal noise for the human visual system (HVS) is used to calculate a signal-to-noise ratio (SNR) related to object detectability. Other inputs to the SNR calculation are the detective quantum efficiency (DQE) and the modulation transfer function (MTF) of the imaging system. Besides threshold detectability, the model predicts the equivalent perception dose ratio (EPDR), which is the fraction of the screen film exposure for which the digital detector provides equal detectability. Images of a contrast-detail phantom are obtained with the digital detector at dose levels corresponding to 27%, 41%, 63% and 100% of the dose used for screen-film. The images are used in a four-alternative forced choice (4-AFC) observer perception study in order to measure threshold detectability. A statistically significant improvement in contrast detectability is measured with the digital detector at 100% and 63% of the screen-film dose. There is no statistical difference between screen-film and digital at 41% of the dose. On average, the experimental EPDR is 44%, which agrees well with the model prediction of 40%. PMID- 11008964 TI - Dental radiography: tooth enamel EPR dose assessment from Rando phantom measurements. AB - Electron paramagnetic resonance dosimetry of tooth enamel is now established as a suitable method for individual dose reconstruction following radiation accidents. The accuracy of the method is limited by some confounding factors, among which is the dose received due to medical x-ray irradiation. In the present paper the EPR response of tooth enamel to endoral examination was experimentally evaluated using an anthropomorphic phantom. The dose to enamel for a single exposure of a typical dental examination performed with a new x-ray generation unit working at 65 kVp gave rise to a CO2- signal of intensity similar to that induced by a dose of about 2 mGy of 60Co. EPR measurements were performed on the entire tooth with no attempt to separate buccal and lingual components. Also the dose to enamel for an orthopantomography exam was estimated. It was derived from TLD measurements as equivalent to 0.2 mGy of 60Co. In view of application to risk assessment analysis, in the present work the value for the ratio of the reference dose at the phantom surface measured with TLD to the dose at the tooth measured with EPR was determined. PMID- 11008966 TI - A study of the directional response of ultraviolet radiometers: I. Practical evaluation and implications for ultraviolet measurement standards. AB - The directional responses of a range of ultraviolet radiometers commonly used for irradiance measurements of UVB and UVC have been studied. Radiometers with 24 diffuser/filter combinations were assessed using a deuterium source, and three different diffuser/filter designs were assessed using a monochromatic source. The directional responses of the radiometers have been calculated and expressed in terms of figures of merit similar to those described for (photopic) illuminance meters in BS 667 and CIE 69. Those radiometers that performed best for the measurement of both small and extended sources of UVB and UVC had raised PTFE diffusers. We conclude that UV radiometers with a directional response error f2 < 10% are readily available commercially, and that it would be appropriate for future ultraviolet standards to set an upper limit of 5% on f2. This would ensure that the overall uncertainty in irradiance measurements of extended ultraviolet sources is not dominated by the error in the directional response of the radiometer. PMID- 11008965 TI - An evaluation of near infrared spectroscopy and cryospectrophotometry estimates of haemoglobin oxygen saturation in a rodent mammary tumour model. AB - Haemoglobin oxygen saturation in subcutaneous rat mammary tumours was measured using near infrared spectroscopy (NIRS) in vivo and in rapidly frozen sections from the same tumours using cryospectrophotometry, which reports oxygen saturation in individual blood vessels to depths of 4 mm from the tissue surface. Measurements were performed on tumours while animals breathed either room air or carbogen. In five of nine tumours, the average saturation calculated from cryospectrophotometric measurements agreed with that determined from NIRS to within 13%, and in four of these five tumours agreement was 5% or better. In the remaining four of nine tumours, where agreement was poor, the volume-averaged saturations estimated from NIRS were consistently higher than those calculated from cryospectrophotometry. Monte Carlo simulations demonstrated that the depth of tissue probed by NIRS was significantly greater than that sampled by cryospectrophotometry. Analysis of the frequency of severely hypoxic vessels showed that when NIRS reported a saturation of approximately 70% or higher, the fraction of tumour vessels with saturations less than 10% was limited to 0.06 or less. Sensitivity and specificity analysis suggests that NIRS and NIRS imaging may identify clinically relevant hypoxia, even when its spatial extent is below the resolution limit of the NIRS technique. PMID- 11008967 TI - A study of the directional response of ultraviolet radiometers: II. Implications for ultraviolet phototherapy derived from computer simulations. AB - A theoretical model has been used to simulate irradiances for ultraviolet (UV) phototherapy cabinets and other sources. The accuracy of the simulation results has been checked by comparison with experimental measurements. The simulations have been used to study the influence of different factors on UV phototherapy exposure and to develop recommendations for the operation and calibration of phototherapy cabinets. Many radiometers used in the evaluation of skin doses have input optics with directional responses that are not proportional to the cosine of the angle of incidence for the UV radiation. Data on radiometer directional responses have been incorporated into the simulations, which show that the poor directional responses for some radiometers currently in use will give errors of 20-50% in the assessment of irradiance. The influence of lamp source geometries employed for radiometer calibration has been investigated. UV phototherapy dosimetry commonly uses a spectroradiometer and a radiometer in the transfer of irradiance calibrations from a small standard UV lamp to a large-area source with a different UV spectrum. Recommendations are given on the range of acceptability for radiometer directional responses and a method is described for determining whether these are fulfilled. Recommendations are made on the techniques that should be used for calibration. PMID- 11008968 TI - Three-dimensional BANG gel dosimetry in conformal carbon ion radiotherapy. AB - In this study we applied BANG polymer-gel dosimetry using magnetic resonance imaging (MRI) to densely ionizing radiation such as carbon ion beams. BANG polymer gels were irradiated with a quadratic field of monoenergetic 12C ions at different beam energies in the range of 135 MeV u(-1) to 410 MeV u(-1). They were irradiated at the radiotherapy facility of the GSI, Darmstadt, Germany. Our object was to examine the saturation effect for densely ionizing radiation that occurs at high values of linear energy transfer (LET). The examination yielded the first effectiveness values that will be discussed in the following sections. A solid sphere and a hollow sphere were both irradiated with a horizontal pencil beam from the raster scanning facility at energies of 268 MeV u(-1) (solid sphere) and 304 MeV u(-1) (hollow sphere) respectively. MR dosimetry measurements were compared with data from a planning system. As far as quality is concerned, there is good agreement between the measured dose distributions of both samples and the dose maps from the planning software. The measured MR signals cannot be converted into absolute dose, since the relative efficiency is still unknown for mixed radiation fields of primary carbon ions and it is known only to a limited extent for nuclear fragments with different energies from highly energetic photon radiation. Model calculations are in progress in order to facilitate conversions of measured MR signals into dose. PMID- 11008969 TI - A novel tracking technique for the continuous precise measurement of tumour positions in conformal radiotherapy. AB - Changing tumour positions induced by organ motion can impede the full exploitation of the strengths of conformal radiotherapy. The unnecessary irradiation of healthy tissue surrounding the target volume can be the consequence. To overcome this, one should measure tumour positions directly and continuously with high resolution in space and time. We have developed a novel tracking technique which will allow this. The method can also be used to survey and monitor the patient positioning. The proper functioning of our method has been technically demonstrated at PSI with the help of phantom irradiation with protons. Implementation into the clinical environment is now beginning. PMID- 11008970 TI - Influence of sulphuric acid contaminants on Fricke dosimetry. AB - The sulphuric acid used for the preparation of the Fricke dosimeter solution may contain trace impurities that can affect the yield of ferric ions. Two methods, pre-irradiation or oxidation with hydrogen peroxide, have been proposed to reduce the influence of these impurities. Fricke users sometimes omit this treatment. In the present work Fricke solutions prepared from six different brands and qualities of sulphuric acid were compared in order to study any influences of the acid on the ferric ion yield. It was shown that the use of analytical grade sulphuric acid from one manufacturer resulted in a reduction of the ferric ion yield of about 5% at an absorbed dose of approximately 20 Gy. If this solution were to be used for an absolute dose determination together with epsilon(m) G values from the literature the absorbed dose would be underestimated by the same amount. PMID- 11008971 TI - Hypoproteinemia predicts acute respiratory distress syndrome development, weight gain, and death in patients with sepsis. Ibuprofen in Sepsis Study Group. AB - OBJECTIVE: Starling's equation indicates that reduced oncotic pressure gradients will favor edema formation, and the current consensus definition of acute respiratory distress syndrome (ARDS) excludes only the hydrostatic pressure contribution. We hypothesized that low serum total protein levels might correlate with the likelihood of ARDS in at-risk patients because serum total protein is the chief determinant of oncotic pressure in humans. DESIGN: Regression analysis to compare outcomes in patients with low serum total protein levels with outcomes in patients with normal serum total protein levels with respect to weight change, development of ARDS, and mortality. SETTING: Intensive care units (ICUs) of seven clinical centers in North America. PATIENTS: A total of 455 ICU patients who met consensus criteria for severe sepsis (178 of whom developed ARDS) from a recently completed prospective clinical trial. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: We found that 92% of the patients developing ARDS had low or borderline serum total protein levels (<6 g/dL). Logistic and multiple regression analyses confirmed that of 18 clinical variables, initial serum total protein level and protein change over time were the most statistically significant predictors of weight gain, prolonged mechanical ventilation, ARDS development, and mortality in the study population. This correlation remained significant after adjustment for the other major predictors of outcome present at baseline (ie, Acute Physiology and Chronic Health Evaluation II score). CONCLUSIONS: Hypoproteinemia is significantly correlated with fluid retention and weight gain, development of ARDS and poor respiratory outcome, and mortality in patients with sepsis. Prospective, randomized trials of serum protein manipulation are needed to establish whether there is a cause-effect relationship to this association. PMID- 11008972 TI - A new approach to Advance Directives. AB - OBJECTIVE: To martial arguments for listing simplified Advance Directives on the Medicare card. DESIGN AND MAIN RESULTS: Literature review shows that 90% of patients do not have advance directives, that patients and doctors are both remiss in discussing end-of-life issues, and that Medicare, insurance companies, and hospitals do little to remedy this lapse. CONCLUSION: A case is made for listing simplified Advance Directives on the Medicare card. PMID- 11008973 TI - Phase I trial of the recombinant soluble complement receptor 1 in acute lung injury and acute respiratory distress syndrome. AB - OBJECTIVE: To determine the safety, pharmacokinetics, biological effects, and immunogenicity of recombinant soluble complement receptor 1 (TP10) in patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). DESIGN: Open label, ascending dosage, phase I trial. SETTING: Two academic teaching hospitals. PATIENTS: A total of 24 patients diagnosed with ALI/ARDS. INTERVENTION: A single, 30-min intravenous infusion of 0.1, 0.3, 1, 3, or 10 mg/kg TP10. MEASUREMENTS AND MAIN RESULTS: Serum levels of TP10 increased in proportion to the dose. Mean variable estimates (+/-SD) were half-life of disposition 69.7 +/- 39.7 hrs, plasma clearance 2.39 +/- 1.32 mL/hr/kg, and volume of distribution 190.6 +/- 135.0 mL/kg. Inhibition of complement activity, measured by CH50, was significant for the interaction of dose and time (p = .024). The C3a levels demonstrated a trend for dose which did not reach statistical significance (p = .090) and soluble C5b-9 levels were significant only for dose (p = .023). As expected by the proposed physiologic mechanism, C4a levels were not affected by TP10, dose, or time. The overall mortality rate was 33%. Neither the type nor the frequency rate of specific adverse events were substantially different between dose groups. Seven adverse events in four patients were thought to be possibly related to TP10. CONCLUSIONS: TP10 has a half-life of approximately 70 hrs and at doses > or =1 mg/kg, significantly inhibits complement activity at the levels of C3 and C5 in patients with ALI/ARDS. Complement inhibition was more prolonged over time with TP10 doses of 3 and 10 mg/kg. TP10 appears to be safe at the doses tested. Further studies will be required to completely assess the impact of TP10 on pathophysiology and clinical outcome in patients with ALI/ARDS. PMID- 11008974 TI - Impact of colony-stimulating factor therapy on clinical outcome and frequency rate of nosocomial infections in intensive care unit neutropenic patients. AB - OBJECTIVES: To determine whether the use of recombinant human granulocyte colony stimulating factor (G-CSF, filgrastim) reduces the mortality rate and the frequency rate of nosocomial infections in neutropenic patients requiring intensive care unit (ICU) admission. DESIGN: Retrospective consecutive case series analysis. SETTING: Medical ICU of a teaching hospital. PATIENTS: We compared two groups of patients, according to whether or not they received G-CSF. In the ICU, 28 leukopenic patients received filgrastim (5 microg of body weight per day intravenously). In all these patients, G-CSF was continued until recovery from leukopenia, defined as a leukocyte count >1,000/mm3. A total of 33 ICU leukopenic patients did not receive G-CSF. End points included leukocyte count, bone marrow recovery, frequency of ICU nosocomial infections (pneumonia, urinary tract, and catheter-related infections), and mortality rate. MEASUREMENTS AND MAIN RESULTS: There were no differences in number of patients who recovered from leukopenia or in whom blood leukocyte count increased. Nosocomial infections occurred in the same percentage in both groups. The percentage of patients who died was identical in both groups. The percentage of patients with and without filgrastim therapy who recovered from leukopenia but died, was 86% and 78%, respectively. CONCLUSION: In the ICU, clinical outcome of neutropenic patients was not changed by G-CSF therapy. It is possible that G-CSF therapy may not be helpful in improving the ICU clinical outcome of neutropenic patients. Additional controlled studies designed to address this question are warranted. PMID- 11008975 TI - Pulmonary endothelial permeability and circulating neutrophil-endothelial markers in patients undergoing esophagogastrectomy. AB - OBJECTIVE: Esophagogastrectomy is an established surgical treatment for esophageal malignancy. The postoperative period may be complicated by the development of acute lung injury syndromes and thus, may provide a useful model in which to study the early pathogenic mechanisms of inflammatory lung injury. DESIGN: Open, prospective study. SETTING: High dependency and intensive therapy units. PATIENTS: Eight healthy male volunteers and 20 patients in the early postoperative period INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The lung protein accumulation index (PAI) of radiolabeled transferrin was determined by using a portable, double-isotope system. The following circulating inflammatory markers-thought to reflect neutrophil-endothelial activation and injury including circulating neutrophil elastase-soluble L-, E-, and P-selectins and thrombomodulin and von Willebrand factor antigen were assayed from venous blood samples The PAI for healthy volunteers was median -0.5 (range, -1.73 to 0.27) x 10(-3)/min and for patients undergoing esophagogastrectomy -0.005 (range, -1.53 to 2.28) x 10(-3)/min. There was no statistical difference between the two groups. In the postesophagogastrectomy group, a significant elevation in circulating levels of neutrophil elastase, soluble P- and E-selectin, thrombomodulin, and von Willebrand factor antigen were observed relative to the control group but only circulating plasma elastase demonstrated a significant correlation with the PAI (r2 = .23, p =.03). CONCLUSIONS: The data suggest patients undergoing esophagogastrectomy develop a inflammatory response but this is not a surrogate of permeability and other factors are likely to determine persistent injury to the alveolar-capillary barrier function in this patient group. PMID- 11008976 TI - Role of interleukin-10 on hyporesponsiveness of endotoxin during surgery. AB - OBJECTIVE: To examine whether surgical stress causes blood cells to lose their responsiveness to endotoxin during surgery. DESIGN: Prospective case series. SETTING: A university hospital. PATIENTS: Sixteen volunteers classified as American Society of Anesthesiologists physical status I-II who were scheduled for elective distal partial gastrectomy. INTERVENTIONS: We studied nine patients who underwent elective distal partial gastrectomy. Blood samples for tumor necrosis factor (TNF) and interleukin (IL)-10 assay were obtained before anesthesia, preincision, 2 hrs and 4 hrs postincision, postextubation, and 24 hrs postincision. The rest of each blood sample was diluted with 5 volumes of endotoxin-free saline, incubated for 4 hrs in the presence of lipopolysaccharide (LPS), centrifuged to remove cells, and assayed for TNF. In another seven patients, antihuman IL-10 antibody was added into the diluted whole blood sample before LPS stimulation. MEASUREMENTS AND MAIN RESULTS: TNF activity was not detected in the blood of any patient throughout the study. In contrast, plasma cortisol and IL-10 levels increased rapidly during surgery (p < .01, p < .05, respectively). LPS-induced TNF activity in whole blood decreased significantly during surgery (p < .01) and recovered to control levels by 24 hrs postincision. The peak suppression of LPS-induced TNF and the peak value of plasma IL-10 levels occurred postextubation. Treatment with anti-IL-10 antibody partially restored the ability of LPS to induce TNF activity postextubation (p < .05). CONCLUSIONS: Surgical trauma rapidly induces a transient hyporesponsiveness of blood cells to endotoxin. Plasma IL-10, which increases during surgery, participates in this hyporesponsiveness. PMID- 11008977 TI - Usefulness of procalcitonin for diagnosis of infection in cardiac surgical patients. AB - OBJECTIVE: To determine the value of procalcitonin (PCT) as a marker of postoperative infection after cardiac surgery. DESIGN: A prospective single institution three phase study. SETTING: University cardiac surgical intensive care unit (31 beds). PATIENTS: Phase 1: To determine the normal perioperative kinetics of PCT, 20 consecutive patients undergoing elective cardiac surgery with cardiopulmonary bypass were included. Phase 2: To determine whether PCT may be useful for diagnosis of postoperative infection, 97 consecutive patients with suspected infection were included. Phase 3: To determine the ability of PCT to differentiate patients with septic shock from those with cardiogenic shock, 26 patients with postoperative circulatory failure were compared. MEASUREMENTS AND MAIN RESULTS: Phase 1: Serum samples were drawn for PCT determination after induction of anesthesia (baseline), at the end of surgery, and daily until postoperative day (POD) 8. Baseline serum PCT concentration was 0.17 +/- 0.08 ng/mL (mean +/- SD). Serum PCT increased after cardiac surgery with a peak on POD 1 (1.08 +/- 1.36). Serum PCT returned to normal range on POD 3 and remained stable thereafter. Phase 2: In patients with suspected infection, serum PCT was measured at the same time of C-reactive protein (CRP) and bacteriologic samples. Among the 97 included patients, 54 were infected with pneumonia (n = 17), bacteremia (n = 16), mediastinitis (n = 9), or septic shock (n = 12). In the 43 remaining patients, infection was excluded by microbiological examinations. In noninfected patients, serum PCT concentration was 0.41 +/- 0.36 ng/mL (range, 0.08-1.67 ng/mL). Serum PCT concentration was markedly higher in patients with septic shock (96.98 +/- 119.61 ng/mL). Moderate increase in serum PCT concentration occurred during pneumonia (4.85 +/-3.31 ng/mL) and bacteremia (3.57 +/- 2.98 ng/mL). Serum PCT concentration remained low during mediastinitis (0.80 +/- 0.58 ng/mL). Five patients with mediastinitis, two patients with bacteremia, and one patient with pneumonia had serum PCT concentrations of <1 ng/mL. These eight patients were administered antibiotics previously and serum PCT was measured during a therapeutic antibiotic window. For prediction of infection by PCT, the best cutoff value was 1 ng/mL, with sensitivity 85%, specificity 95%, positive predictive value 96%, and negative predictive value 84%. Serum CRP was high in all patients without intergroup difference. For prediction of infection by CRP, a value of 50 mg/L was sensitive (84%) but poorly specific (40%). Comparing the area under the receiver operating characteristic curves, PCT was better than CRP for diagnosis of postoperative sepsis (0.82 for PCT vs. 0.68 for CRP). Phase 3: Serum PCT concentration was significantly higher in patients with septic shock than in those with cardiogenic shock (96.98 +/- 119.61 ng/mL vs. 11.30 +/- 12.3 ng/mL). For discrimination between septic and cardiogenic shock, the best cutoff value was 10 ng/mL, with sensitivity of 100% and specificity of 62%. CONCLUSION: Cardiac surgery with cardiopulmonary bypass influences serum PCT concentration with a peak on POD 1. In the presence of fever, PCT is a reliable marker for diagnosis of infection after cardiac surgery, except in patients who previously received antibiotics. PCT was more relevant than CRP for diagnosis of postoperative infection. During a postoperative circulatory failure, a serum PCT concentration >10 ng/mL is highly indicative of a septic shock. PMID- 11008978 TI - Randomized trial of the use of heliox as a driving gas for updraft nebulization of bronchodilators in the emergent treatment of acute exacerbations of chronic obstructive pulmonary disease. AB - OBJECTIVE: To determine whether the bronchodilator effects of albuterol and ipratropium bromide are greater if updraft nebulization is driven by 80% helium and 20% oxygen (HELIOX) than if driven by compressed room air (AIR) during the treatment of an acute exacerbations of chronic obstructive pulmonary disease (COPD). SETTING: The emergency department of a 750-bed inner-city community hospital. METHODS: Over a 12-month period, a convenience sample of 50 normoxic patients presenting with signs and symptoms of an acute exacerbation of COPD were prospectively randomized to receive either HELIOX or AIR as the driving gas for updraft nebulization of a mixture of albuterol 2.5 mg and ipratropium bromide 0.5 mg. Additional aerosol treatments with albuterol 2.5 mg were given at 20, 40, and 120 mins after randomization using the assigned gas. Spirometry was obtained while breathing room air before the first treatment (baseline) and at 1 hr and 2 hrs after the initiation of treatment. The primary measure of efficacy was the change in percent of predicted forced expiratory volume in 1 sec (FEV1) over the treatment period. A secondary measure of efficacy was the change in percentage of predicted forced expiratory flow after 25% to 75% of vital capacity had been expelled (FEF25-75). RESULTS: Twenty-five patients were randomized to each treatment group. Three patients (1 HELIOX, 2 AIR) were unable to complete the study. The baseline FEV1was 44% (95% confidence interval, 35% to 52%) of predicted in the HELIOX group and 39 (31% to 46%) of predicted in the AIR group. There were no adverse outcomes observed in either the HELIOX group or the AIR group. There were no significant differences in the change of FEV1 between the two groups by either the 1 hr or 2 hr time point (1 hr, HELIOX + 10% [7% to 13%], AIR + 9% [5% to 13%]; 2 hr HELIOX + 10% [6% to 15%], AIR + 10% [6% to 14%]). The improvement in FEF25-75 was significantly greater in the HELIOX group than in the AIR group at both the 1 hr time point (HELIOX + 14% [7% to 22%] vs. AIR + 7% [3% to 10%], p = .05) and at the 2 hr time point (HELIOX + 15% [8% to 21%] vs. AIR + 7% [4% to 11%], p = .05). CONCLUSION: Use of HELIOX as a driving gas for the updraft nebulization of bronchodilators during the first 2 hrs of treatment of an acute COPD exacerbation failed to improve FEV1 faster than the use of AIR. The faster improvement in FEF25-75 during the first 2 hrs of treatment was small and of uncertain clinical significance. PMID- 11008979 TI - Venous blood flow measurement by determination of change in venous hemoglobin saturation. AB - OBJECTIVE: To evaluate a method of measuring venous blood flow in vitro by using the Fick principle and change in saturation of venous blood and to apply the method to the clinical measurement of hepatic blood flow. DESIGN: Experimental study using an in vitro model and clinical study for critically ill patients. SETTING: Department of Anesthesiology and Intensive Care Medicine in Osaka City University Medical School. MODEL: Human blood deoxygenated by bubbling of nitrogen was circulated in a closed circuit at 10-120 mL/min by a roller pump. A fiber optic sensor was attached to the circuit for continuous monitoring of hemoglobin saturation. PATIENTS: Eight critically ill patients, aged 54.3 +/- 15.1 yrs, were studied. INTERVENTIONS: Hemoglobin saturation was changed in the in vitro study by the injection of 0.2 mL of oxygenated blood. Signals from the optical fiber were analogue-digital converted and recorded in a computer. In the clinical study, an oximetry catheter was inserted into the inferior vena cava (IVC) via the femoral vein. Arterial blood (10 mL) was drawn from the radial artery, and injected into the IVC. The changes in oxygen saturation in the venous blood were recorded. MEASUREMENTS AND MAIN RESULTS: Blood flow was calculated using the Fick principle, assuming that all the injected blood passes through the sensor. In vitro estimation of blood flow was well correlated with the actual flow (r2 = .94). IVC blood flow was measured above and below the merging point of the hepatic vein. The difference of the two values was assumed to represent hepatic blood flow. IVC blood flow was calculated by the same method as for the in vitro study. The blood flows in the IVC above and below the anastomosis with the hepatic vein were 2.82 +/-0.56 (SD) Umin and 1.96 +/- 0.61 (SD) L/min. Average estimated hepatic blood flow was 0.86 L/min (range, 0.34-1.75 Umin). CONCLUSION: We examined the accuracy and reliability of this new method in the present in vitro study. This method may be clinically useful for measuring hepatic blood flow. PMID- 11008980 TI - Noninvasive continuous positive airway pressure in neutropenic patients with acute respiratory failure requiring intensive care unit admission. AB - OBJECTIVE: To evaluate the tolerance and the efficacy of noninvasive continuous positive airway pressure (CPAP) in severe acute respiratory failure occurring in intensive care unit (ICU) neutropenic patients with hematologic malignancies, and to establish predictive variables of efficacy of this method. DESIGN: Prospective study over a 5-yr period. SETTING: Hematologic and medical intensive care unit of a teaching hospital. METHODS: Among 129 neutropenic patients admitted to the ICU, 64 patients presented with febrile acute hypoxemic normocapnic respiratory failure (PaO2/FIO2 ratio <200) and were enrolled. In addition to standard therapy, patients received CPAP with a facial mask. The initial settings of the CPAP were 6 cm H2O positive end-expiratory pressure and FIO2 0.8 (80%). Physiologic measurements were performed at the end of 45 mins of ventilation with first adjustments. CPAP was used with a sequential mode (45 mins/3 hrs). CPAP was efficient if intubation was avoided. RESULTS: The setting of CPAP, after adjustments, was as follows: positive end-expiratory pressure 7 +/- 1 cm H2O and FIO2 0.7 +/- 0.1 (70% +/- 10%). For the 64 patients, CPAP was administered for a total of 6 +/- 2 hrs during the first 24 hrs. The mean duration of CPAP was 7 +/- 3 days. A reduction in respiratory rate to less than 25 breaths/min was achieved in 53% of patients. PaO2/FIO2 ratio increased from 128 +/- 32 to 218 +/- 28. CPAP was successful in avoiding endotracheal intubation in 16/64 patients. A total of 16 responders and four nonresponders survived. Hepatic failure was a criterion indicating the failure of CPAP: 1/16 vs. 26/48 (p = .001). In multivariate analysis, two variables were predictive of failure of CPAP: Simplified Acute Physiology Score II (58 +/- 14 vs. 41 +/- 11) and a hepatic failure at the entry into the study. CONCLUSION: CPAP was efficient in 25% of cases. All the responders survived. This noninvasive method was used as a way to avoid mechanical ventilation, which is well correlated with a poor prognosis in neutropenic ICU patients. Further controlled studies are needed to confirm the efficacy of noninvasive CPAP and to evaluate the most appropriate selection of immunocompromised patients. PMID- 11008981 TI - The relationship between visceral ischemia, proinflammatory cytokines, and organ injury in patients undergoing thoracoabdominal aortic aneurysm repair. AB - OBJECTIVES: Plasma proinflammatory, anti-inflammatory cytokine, and soluble tumor necrosis factor (TNF) receptor concentrations were examined in hospitalized patients after abdominal and thoracoabdominal aortic aneurysm (TAAA) repair, with and without left atrial femoral bypass. Changes in plasma cytokine concentrations were related to the duration of visceral ischemia and the frequency rate of postoperative, single, or multiple system organ dysfunction (MSOD). DESIGN: Prospective, observational study. SETTING: Two academic referral centers in the United States and The Netherlands. PATIENTS: We included 16 patients undergoing TAAA repair without left atrial femoral bypass, 12 patients undergoing TAAA repair with left atrial femoral bypass, and nine patients undergoing infrarenal aortic aneurysm repair. MEASUREMENTS AND MAIN RESULTS: Timed, arterial blood sampling for proinflammatory and anti-inflammatory cytokine and soluble TNF receptor concentrations (p55 and p75), and prospective assessment of postoperative single and MSOD. Plasma appearance of TNF-alpha, interleukin (IL) 6, IL-8, and IL-10 peaked 1 to 4 hrs after TAAA repair, and concentrations were significantly elevated compared with infrarenal abdominal aortic aneurysm repair (p < .05). Left atrial femoral bypass significantly reduced the duration of visceral ischemia (p < .05) and the systemic TNF-alpha, p75, and IL-10 responses (p < .05). Plasma TNF-alpha concentrations >150 pg/mL were more common in patients with extended visceral ischemia times (>40 mins). Additionally, patients with early peak TNF-alpha concentrations >150 pg/mL and IL-6 levels >1,000 pg/mL developed MSOD more frequently than patients without these elevated plasma cytokine levels (both p < .05). CONCLUSIONS: Thoracoabdominal aortic aneurysm repair results in the increased plasma appearance of TNF-alpha, IL-6, IL-8, IL 10, and shed TNF receptors. The frequency and magnitude of postoperative organ dysfunction after TAAA repair is associated with an increased concentration of the cytokines, TNF-alpha, and IL-6 and the increased plasma levels of these cytokines appear to require extended visceral ischemia times. PMID- 11008982 TI - High adenosine plasma concentration as a prognostic index for outcome in patients with septic shock. AB - OBJECTIVE: Sepsis and septic shock are a common cause of mortality in critically ill patients. Many substances have been implicated in the pathophysiology of these syndromes. We postulated that adenosine may be implicated in the sepsis- or septic shock-induced blood pressure failure. Indeed, this nucleoside is a strong endogenous vasodilating agent released by endothelial cells and myocytes under circumstances of metabolic stress, such as during critical illness. DESIGN: A prospective, comparative observational study. SETTING: The adult intensive care unit of a tertiary care university hospital. PATIENTS: We measured adenosine plasma concentration (APC) in patients with severe sepsis (n = 11), in patients with septic shock (n = 14), in patients with hemorrhagic traumatic shock (n = 14), and in 12 healthy volunteers. APC was evaluated every 12 hrs over 3 days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: At study entry, we found that APC was higher in patients with septic shock (mean +/- so = 8.4 +/-3.5 micromol/L) than in patients with hemorrhagic traumatic shock (1.1 +/- 0.6 micromol/L) and controls (0.8 +/- 0.3 micromol/L). Intermediate values (3.9 +/- 1.9 micromol/L) were found in patients with severe sepsis. APC in patients with traumatic shock did not differ from controls. In the course of the hospitalization, for both sepsis and septic shock patients, APC decreased significantly but remained higher than controls 72 hrs after entry into the study. In the septic shock group, APC was significantly higher in the nonsurvivor group (n = 6) than in the survivor group (n = 8), whatever the time of sample collection and assay. CONCLUSIONS: High adenosine plasma concentrations are found in patients with septic shock but not during traumatic shock, or in healthy volunteers. Intermediate values of circulating adenosine are found in patients with severe sepsis. APC may be a prognostic index for outcome in septic patients, with much higher values being found in nonsurvivors. PMID- 11008983 TI - G-protein beta3 subunit gene (GNB3) polymorphism 825C-->T in patients with hypertensive crisis. AB - OBJECTIVE: The polymorphism 825C-->T in exon 10 of the gene GNB3 encoding the beta3 subunit of heterotrimeric guanine nucleotide binding regulatory proteins (G proteins) results in a splicing variant (GNB3-s) in which the nucleotides 498-620 of exon 9 are deleted. The T allele has been shown to be overrepresented in patients with essential hypertension. Because GNB3-s may support the development of severe elevation of blood pressure, we hypothesized that GNB3 825C-->T may be present more frequently in patients with hypertensive crisis. DESIGN: Case control study. SETTING: Department of Emergency Medicine at the University Hospital of Vienna, Vienna, Austria. PATIENTS: A total of 174 patients admitted to an emergency department for treatment of hypertensive crisis diagnosed as suffering from essential hypertension. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were genotyped for the 825C-->T transition in GNB3. An equal number of age- and gender-matched normotensive, healthy individuals served as the control population. The allele frequency of 825C-->T in the GNB3 gene was 0.310 in patients with hypertensive crisis and 0.342 in the control group. There was no difference in genotype distribution and allele frequency between the patients and the age- and gender-matched control group or between the observed prevalence and the occurrence rate expected from the Hardy-Weinberg principle within each group. CONCLUSIONS: GNB3 825C-->T is not associated with the phenotype of hypertensive crisis in patients suffering from essential hypertension. Furthermore, our data do not support the concept that the 825C-->T transition in the GNB3 gene is associated with essential hypertension. PMID- 11008984 TI - Rapid onset of intestinal epithelial and lymphocyte apoptotic cell death in patients with trauma and shock. AB - OBJECTIVE: Apoptosis is a cellular suicide program that can be activated by cell injury or stress. Although a number of laboratory studies have shown that ischemia/reperfusion injury can induce apoptosis, few clinical studies have been performed. The purpose of this study was to determine whether apoptosis is a major mechanism of cell death in intestinal epithelial cells and lymphocytes in patients who sustained trauma, shock, and ischemia/ reperfusion injury. DESIGN: Intestinal tissues were obtained intraoperatively from 10 patients with acute traumatic injuries as a result of motor vehicle collisions or gun shot wounds. A control population consisted of six patients who underwent elective bowel resections. Apoptosis was evaluated by conventional light microscopy, laser scanning confocal microscopy using the nuclear staining dye Hoechst 33342, immunohistochemical staining for active caspase-3, and immunohistochemical staining for cytokeratin 18. SETTING: Academic medical center. PATIENTS: Patients with trauma or elective bowel resections. MEASUREMENTS AND MAIN RESULTS: Extensive focal crypt epithelial and lymphocyte apoptosis were demonstrated by multiple methods of examination in the majority of trauma patients. Trauma patients having the highest injury severity score tended to have the most severe apoptosis. Repeat intestinal samples obtained from two of the trauma patients who had a high degree of apoptosis on initial evaluation were negative for apoptosis at the time of the second operation. Tissue lymphocyte apoptosis was associated with a markedly decreased circulating lymphocyte count in 9 of 10 trauma patients. CONCLUSIONS: Focal apoptosis of intestinal epithelial and lymphoid tissues occurs extremely rapidly after injury. Apoptotic loss of intestinal epithelial cells may compromise bowel wall integrity and be a mechanism for bacterial or endotoxin translocation into the systemic circulation. Apoptosis of lymphocytes may impair immunologic defenses and predispose to infection. PMID- 11008985 TI - No long-term benefit from hypothermia after severe traumatic brain injury with secondary insult in rats. AB - OBJECTIVES: To evaluate the effect of application of transient, moderate hypothermia on outcome after experimental traumatic brain injury (TBI) with a secondary hypoxemic insult. DESIGN: Prospective, randomized study. SETTING: University-based animal research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: All rats were subjected to severe TBI followed by 30 mins of moderate hypoxemia, associated with mild hypotension. Rats were randomized to three groups: a) normothermia (37 degrees C + 0.5 degrees C); b) immediate hypothermia (32 degrees C +/- 0.5 degrees C initiated after trauma, before hypoxemia); and c) delayed hypothermia (32 degrees C +/- 0.5 degrees C after hypoxemia). The brain temperature was controlled for 4 hrs after TBI and hypoxemia. MEASUREMENTS AND MAIN RESULTS: Animals were evaluated after TBI for motor and cognitive performance using beam balance (days 1-5 after TBI), beam walking (days 1-5 after TBI), and Morris Water Maze (days 14-18 after TBI) assessments. On day 21 after TBI, rats were perfused with paraformaldehyde and brains were histologically evaluated for lesion volume and hippocampal neuron counts. All three groups showed marked deficits in beam balance, beam walking, and Morris Water Maze performance. However, these deficits did not differ between groups. There was no difference in lesion volume between groups. All animals had significant hippocampal neuronal loss on the side ipsilateral to injury, but this loss was similar between groups. CONCLUSIONS: In this rat model of severe TBI with secondary insult, moderate hypothermia for 4 hrs posttrauma failed to improve motor function, cognitive function, lesion volume or hippocampal neuronal survival. Combination therapies may be necessary in this difficult setting. PMID- 11008986 TI - C1 inhibitor prevents capillary leakage after thermal trauma. AB - OBJECTIVE: In burned patients, activation of the complement and clotting systems is suggested to play an important role in the development of the capillary leak syndrome and inflammatory tissue destruction. In an animal model of thermal trauma, the possible protective effect of C1 inhibitor (C1Inh), a major control protein of both the complement and clotting systems, was investigated. DESIGN: Prospective, controlled study. SETTING: Animal model. SUBJECTS: Healthy pigs weighing 30 kg. INTERVENTIONS: Pigs were scalded for 25 secs with 75 degrees C hot water to achieve a 30% total body surface deep partial-thickness burn. The treatment group (n = 8) received C1Inh concentrate at an initial dose of 100 units/kg body weight immediately after thermal trauma, followed by three further applications every 12 hrs. Two control groups included animals that were either scalded (n = 8) or not scalded (n = 7) and treated with lactated Ringer's solution. MEASUREMENTS: Before and at various time points after trauma blood samples were analyzed for complement activation (APH50, CH50, SC5b-9, C3). Continuous monitoring of hemodynamic variables was performed and postmortem histologic examination of specimens from lung, heart, liver, kidney, stomach, duodenum, jejunum, ileum, and colon was carried out. Aseptically collected mesenteric lymph nodes were pooled and screened for bacterial translocation. For evaluation of the burn wound, biopsies from defined scalded and not scalded areas were taken daily. As a measure for edema formation, the weight of the animals was recorded every 2 hrs. RESULTS: After C1Inh treatment, which led to a significantly reduced complement activation, the clinical outcome was clearly improved, as indicated by vital signs and as demonstrated by reduced edema formation. Treated animals presented a diminished bacterial translocation. Pathologic alterations were clearly diminished in the burned skin, in shock related organs, and in the intestines. CONCLUSION: Application of C1Inh appears to be an effective means to prevent capillary leakage and inflammatory tissue destruction after thermal trauma. PMID- 11008987 TI - Dynamic study of the distribution of microcirculatory blood flow in multiple splanchnic organs in septic shock. AB - OBJECTIVES: To study dynamic distribution of microcirculatory blood flow in multiple splanchnic organs during septic shock; to test the hypothesis that changes in microcirculatory blood flow in splanchnic organs correlate with changes in regional flow during septic shock. DESIGN: A prospective, controlled, animal study. SETTING: Animal laboratory in a university medical center. SUBJECTS: Nine anesthetized and mechanically ventilated domestic pigs. INTERVENTIONS: Systemic flow (cardiac output) was measured with thermodilution and regional (superior mesenteric artery) flow with transit time flowmetry. Local blood flow (microcirculatory flow) was continuously measured in splanchnic organs (gastric, jejunal, and colon mucosa, liver, and pancreas) and the kidney with multichannel laser Doppler flowmetry. Septic shock was induced with fecal peritonitis. After 240 mins of sepsis, intravenous fluids were administered to alter hypodynamic shock to hyperdynamic septic shock. MEASUREMENTS AND MAIN RESULTS: In this severe septic shock model, systemic and regional flows decreased by approximately 50% during the first 240 mins. Similar reductions were recorded in microcirculatory flow in the mucosa of the stomach (-41%; p < .001) and colon (-47%; p < .001). In the jejunal mucosa, on the other hand, flow remained virtually unchanged. Microcirculatory flow was also significantly decreased in the liver (-49%; p < .001), pancreas (-56%; p < .001), and kidney (-44%; p < .001). Administration of intravenous fluids at 240 mins was followed by three fold increases in systemic and regional flows (approximately 70% above baseline). In the jejunal mucosa, flow also increased significantly above baseline (42%; p < .001), whereas in the stomach and the colon, it barely reached baseline. Kidney blood flow increased to baseline, whereas pancreas and liver flows remained 26% (p < .05) and 34% (p < .001), respectively, below baseline. CONCLUSION: Changes in microcirculatory blood flow in the splanchnic organs are heterogeneous, both in early hypodynamic and in hyperdynamic septic shock, and cannot be predicted from changes in systemic or regional flows. Microcirculatory blood flow in the jejunal mucosa remains constant during early septic shock, whereas pancreatic blood flow decreases significantly more than regional flow. PMID- 11008988 TI - L-arginine attenuates trauma-hemorrhage-induced liver injury. AB - OBJECTIVES: Liver injury is common after trauma-hemorrhage for which the underlying mechanism is not clear. Although administration of the essential amino acid L-arginine has been reported to restore the depressed cardiovascular functions and cell-mediated immune responses after trauma-hemorrhage, it remains unknown whether L-arginine protects against liver injury under those conditions. DESIGN: A prospective, controlled animal study. SETTING: A university research laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: Rats underwent sham operation or laparotomy and were bled to and maintained at a mean arterial blood pressure of 40 mm Hg until 40% of the maximum shed blood volume was returned in the form of lactated Ringer's solution. Hemorrhaged rats were then resuscitated with lactated Ringer's solution, four times the maximum shed blood volume over 1 hr. During resuscitation, animals received either 300 mg/kg of L arginine or saline (vehicle) intravenously. At 3 and 5 hrs after resuscitation, rats were killed, blood was obtained, and the liver was fixed for histology (hematoxylin & eosin staining). Plasma glutathione S-transferase (a marker of liver damage), L-arginine, citrulline, and ornithine concentrations were assessed. MEASUREMENTS AND MAIN RESULTS: The increased concentrations of plasma glutathione S-transferase observed in vehicle-treated hemorrhage animals were normalized with L-arginine treatment at 5 hrs after resuscitation. Moreover, the histology indicated that L-arginine prevented liver edema and neutrophil infiltration after trauma-hemorrhage. Plasma L-arginine and citrulline were increased in L-arginine-treated rats. CONCLUSIONS: Because citrulline is a by product of nitric oxide generation by nitric oxide synthase from L-arginine, this amino acid may be a useful adjunct for preventing hepatic injury after trauma hemorrhage via endothelial derived nitric oxide production. PMID- 11008989 TI - Nuclear factor-kappaB activation in mouse lung lavage cells in response to Streptococcus pneumoniae pulmonary infection. AB - OBJECTIVES: To assess the state and activation kinetics of the nuclear transcription regulatory protein nuclear factor-kappB (NF-kappaB) in lung lavage cells in a murine pneumococcal pneumonia model and to determine how the virulence of the infecting organisms altered the activation state of NF-kappaB. DESIGN: Experimental, comparative study of three Streptococcus pneumoniae strains that induced three distinct pulmonary diseases. SETTING: Experimental laboratory in a university-based medical center. SUBJECTS: Female BALB/cby mice, 8-10 wks of age. INTERVENTIONS: We randomly divided the mice into the following five groups: a) the control group; b) animals infected by virulent encapsulated S. pneumoniae P4241 strain; c) animals infected by avirulent encapsulated S. pneumoniae P15986 strain; d) animals infected by avirulent unencapsulated S. pneumoniae R6 strain; e) animals infected by virulent lysed S. pneumoniae P4241 strain. Animals were anesthetized and infected by intratracheal delivery of 4 x 10(5) colony-forming units (CFU) of S. pneumoniae per mouse or bacterial components equivalent to 4 x 10(5) CFU for lysed S. pneumoniae challenge. After intratracheal challenge with virulent encapsulated strain P4241, mice developed acute pneumonia, became bacteremic, and died within 3 to 5 days. None of the mice infected with the avirulent encapsulated strain P15986 or the avirulent unencapsulated strain R6 died. After collection of lung lavage cells and nuclear extraction, NF-kappaB activation was determined 1 hr, 4 hrs, 6 hrs and 24 hrs after pneumococcal infection. At the same time, pulmonary and blood clearance, bronchoalveolar lavage cells population, and tumor necrosis factor-alpha production were assessed (six mice per time point). MEASUREMENTS AND MAIN RESULTS: NF-kappaB was constitutively expressed within nuclear extracts of lung lavage cells from uninfected control mice. A significant increase in NF-kappaB activation was detected within 1 hr after injection of virulent lysed S. pneumoniae P4241 strain (bacterial components equivalent to 4 x 10(5) CFU), and was still present 24 hrs after the injection. After live pneumococcal challenge, significant NF-kappaB activation was detected within 4 hrs with a peak at 24 hrs. Responses to all three strains (P4241, P15986 and R6) were time-dependent (p < .0001), as NF kappaB activation gradually increased during the first 24 hrs. Moreover, compared with the control uninfected mice, the intensity of the retarded KB oligonucleotide, as determined by densitometry, was increased approximately four- to five-fold and seven-fold in reactions containing nuclear extracts isolated 24 hrs after infection with the avirulent strains P15986 or R6 and the virulent strain P4241, respectively. With the virulent strain P4241, responses were significantly stronger than with the avirulent strains P15986 and R6 (p < .01). Responses were of similar order with avirulent strains P15986 and R6 (p > .05). CONCLUSION: Pulmonary infection by S. pneumoniae induced delayed and time dependent activation of NF-kappaB in mouse lung lavage cells. The degree of NF kappaB activation in lung lavage cells correlated with the virulence of the infecting organisms. Our results suggest that the more severe the infection, the higher the rise in NF-kappaB. PMID- 11008991 TI - CO2 reactivity and brain oxygen pressure monitoring in severe head injury. AB - OBJECTIVE: To investigate the effect of hyperventilation on cerebral oxygenation after severe head injury. DESIGN: A prospective, observational study. SETTING: Neurointensive care unit at a university hospital. PATIENTS: A total of 90 patients with severe head injury (Glasgow Coma Scale score < or =8), in whom continuous monitoring of brain tissue oxygen pressure (PbrO2) was performed as a measure of cerebral oxygenation. INTERVENTIONS: Arterial PCO2 was decreased each day over a 5-day period for 15 mins by increasing minute volume on the ventilator setting to 20% above baseline. Arterial blood gas analysis was performed before and after changing ventilator settings. Multimodality monitoring, including PbrO2, was performed in all patients. Absolute and relative PbrO2/PaCO2 reactivity was calculated. Outcome at 6 months was evaluated according to the Glasgow Outcome Scale. MEASUREMENTS AND MAIN RESULTS: Effective hyperventilation, defined by a decrease of PaCO2 > or =2 torr (0.27 kPa), was obtained in 218 (84%) of 272 tests performed. Baseline PaCO2 averaged 32.3 +/- 4.5 torr (4.31 +/- 0.60 kPa). Average reduction in PaCO2 was 3.8 +/- 1.7 torr (0.51 +/- 0.23 kPa). PbrO2 decreased by 2.8 +/- 3.7 torr (0.37 +/- 0.49 kPa; p < .001) from a baseline value of 26.5 +/- 11.6 torr (3.53 +/- 1.55 kPa). PbrO2/PaCO2 reactivity was low on day 1 (0.8 +/- 2.3 torr [0.11 +/- 0.31 kPa]), increasing on subsequent days to 6.1 +/ 4.4 torr (0.81 +/- 0.59 kPa) on day 5. PbrO2/PaCO2 reactivity on days 1 and 2 was not related to outcome. In later phases in patients with unfavorable outcome, relative reactivity was increased more markedly, reaching statistical significance on day 5. CONCLUSIONS: Increased hyperventilation causes a significant reduction in PbrO2, providing further evidence for possible increased risk of secondary ischemic damage during hyperventilation. The low PbrO2/PaCO2 reactivity on day 1 indicates the decreased responsiveness of cerebral microvascular vessels to PaCO2 changes, caused by generalized vascular narrowing. The increasing PbrO2/PaCO2 reactivity from days 2 to 5 suggests that the risk of compromising cerebral oxygenation by hyperventilation may increase over time. PMID- 11008990 TI - Comparison between selective and nonselective nitric oxide synthase inhibition and phenylephrine in normal and endotoxic swine. AB - OBJECTIVE: To compare the cardiopulmonary and peripheral circulatory effects of the nonselective nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) to the more selective inducible NOS inhibitor S-methylisothiourea (SMT) and to phenylephrine (PE) in endotoxic and normal swine. DESIGN: Prospective, randomized, unblinded study. SETTING: Research laboratory of academic medical center. SUBJECTS: Nonanesthetized, sedated, mechanically ventilated, minimally invasive swine model. INTERVENTIONS: Animals received either lipopolysaccharide (LPS, LPS groups) or equivalent volume of saline (normal groups). LPS animals were further randomized into four groups when mean arterial pressure (MAP) had dropped to <60 mm Hg: the LPS/saline group received saline only; the other groups received either L-NAME, SMT, or PE. These were titrated to elevate MAP by 20-25 mm Hg, and animals were followed for another 3 hrs. Pulmonary artery occlusion pressure was maintained at one to two times baseline with the infusion of saline. Normal groups received the same agents 1 hr after baseline measurements, and drugs were titrated to achieve similar increases in MAP. We measured gastric-arterial PCO2 gradient by tonometry as an index of gastric mucosal perfusion. Left ventricular volumes were determined echocardiographically; right ventricular volumes were determined by a pulmonary arterial catheter equipped with a rapid thermistor. Plasma nitrite/nitrate (NOx) concentrations were measured hourly. MEASUREMENTS AND MAIN RESULTS: In the LPS groups, all agents elevated MAP and systemic vascular resistance similarly. By hr 4, cardiac output had decreased in all groups, but the decrease with L-NAME (35% +/- 16%) occurred earlier (at hr 3) and was larger than the decrease with SMT at hrs 3 and 5 and larger than the decrease with saline at hrs 3 to 5. L-NAME resulted in a larger increase in mean pulmonary arterial pressure (MPAP) when compared with saline (130% +/- 44% vs. 61% +/- 25%; p < .001) and SMT groups (130% vs. 97% +/- 80%; p < .007). Only L-NAME had detrimental effects on right ventricular function as indicated by an increase in right ventricular end systolic volume (54 +/- 10 to 87 +/-6 mL; p < .05) and right ventricular end diastolic volume (90 +/-11 to 128 +/- 18 mL; p < .05). SMT decreased both left ventricular end-systolic volume (10.4 +/- 2 to 7.7 +/- 4 mL; p < .05) and left ventricular end-diastolic volume (18.5 +/- 3 to 14.2 +/- 5 mL; p < .05), indicating improved left ventricular function, whereas L-NAME did not affect left ventricular volumes. Both SMT and PE corrected LPS-induced gastric mucosal acidosis, but L-NAME did not. We did not detect changes in plasma NOx concentrations in any of LPS groups. In the normal groups, all agents increased MAP without changes in plasma NOx concentrations. L-NAME caused a larger decrease in cardiac output, but the increase in MPAP was higher with SMT. Both NOS inhibitors led to left ventricular dilation, but PE did not. Only L-NAME caused right ventricular dilation. There were no changes in gastric-arterial PCO2 gradient. CONCLUSIONS: In LPS animals, we failed to detect changes in plasma NOx concentrations. Furthermore, for similar increases in MAP, SMT improved gastric mucosal acidosis, had less adverse effects on right ventricular function and MPAP, and may have improved left ventricular function. However, apart from its bene-ficial effects on left ventricular function, SMT was not superior to PE. The results from normal animals indicate that both NOS inhibitors have adverse effects on cardiac function beyond those attributed to increased MAP. PMID- 11008992 TI - Measurements of cortical cellular pH by intracranial tonometer in severe head injury. AB - OBJECTIVE: To evaluate the cortical cellular damage in acute severe head injury, we measured the cortical cellular pH by using an intracranial tonometer made in our institution. DESIGN: Prospective, 3.5-yr data collection. SETTING: University hospital trauma intensive care unit. PATIENTS: Severely head-injured patients (n = 29) with Glasgow Coma Scale score <8. INTERVENTION: Routine emergency neurologic procedure. MEASUREMENTS AND MAIN RESULTS: We made 98 measurements of cortical cellular pH by intracranial tonometer in 29 severely head-injured patients in the acute phase. Each patient's intracranial pressure was recorded, and in 16 patients, the saturation of jugular venous oxygen was monitored. The outcome at 6 months after injury was significantly better in patients having a cortical cellular pH of >7.2 than those with <7.2. The cerebral perfusion pressure and cortical cellular pH correlated significantly (p < .0001). CONCLUSIONS: Our study suggests the usefulness of measurement of cortical cellular pH by intracranial tonometer for evaluating the severity of focal anaerobic cerebral metabolism and predicting patient prognosis. PMID- 11008993 TI - Experience and endocrine stress responses in neonatal and pediatric critical care nurses and physicians. AB - OBJECTIVE: Critical care is a working environment with frequent exposure to stressful events. High levels of psychological stress have been associated with increased prevalence of burnout. Psychological distress acts as a potent trigger of cortisol secretions. We attempted to objectify endocrine stress reactivity. DESIGN: Observational cohort study during two 12-day periods in successive years. SETTING: A tertiary multidisciplinary neonatal and pediatric intensive care unit (33 beds). SUBJECTS: One hundred and twelve nurses and 27 physicians (94% accrual rate). INTERVENTIONS AND MEASUREMENTS: Cortisol determined from salivary samples collected every 2 hrs and after stressful events. Participants recorded the subjective perception of stress with every sample. Endocrine reactions were defined as transient surges in cortisol of >50% and 2.5 nmol/L over the baseline. MAIN RESULTS: During 7,145 working hours, we observed 474 (12.5%) endocrine reactions from 3,781 samples. The mean cortisol increase amounted to 10.6 nmol/L (219%). The mean occurrence rate of endocrine reactions per subject and sample was 0.159 (range, 0-0.43). Although the mean raw cortisol levels were lower in experienced team members (>3 yrs of intensive care vs. <3 yrs, 4.1 vs. 4.95 nmol/L, p < .001), professional experience failed to attenuate the frequency and magnitude of endocrine reactions, except for the subgroup of nurses and physicians with >8 yrs of intensive care experience. A high proportion (71.3%) of endocrine reactions occurred without conscious perception of stress. Unawareness of stress was higher in intensive care nurses (75.1%) than in intermediate care nurses (51.8%, p < .01). CONCLUSIONS: Stress-related cortisol surges occur frequently in neonatal and pediatric critical care staff. Cortisol increases are independent of subjective stress perception. Professional experience does not abate the endocrine stress reactivity. PMID- 11008994 TI - A controlled trial of cardiopulmonary resuscitation training for ethnically diverse parents of infants at high risk for cardiopulmonary arrest. AB - OBJECTIVE: Parents of infants hospitalized in the neonatal intensive care unit are routinely taught cardiopulmonary resuscitation (CPR) as part of the preparation for transition to home. A variety of methods are used to teach CPR knowledge and skills. The purpose of this study was to compare the psychosocial consequences of three different methods of CPR training for parents of infants at high risk for cardiopulmonary arrest. DESIGN, SETTING, AND SUBJECTS: In this prospective, multisite clinical trial, 335 parents and other caretakers of infants hospitalized in the neonatal intensive care unit were followed for 1 yr. Participants were 69% female, 49% Latino/Latina, with a mean age of 30 +/- 8 yrs. INTERVENTION: Subjects initially were randomized to one of four CPR training protocols: a video-only class, an instructor-taught class, an instructor-taught class combined with a social support intervention, and a control group. The social support intervention involved a group discussion after CPR training with regular telephone follow-up over the next 6 months by a nurse. MEASUREMENTS AND MAIN RESULTS: Data on psychosocial adjustment to illness, anxiety, and depression were collected at baseline, 2 wks, and 3 and 6 months. There were significant differences among the groups over time in anxiety (p = .007) and psychosocial adjustment to illness (p = .001). Parents in the CPR-video protocol had significantly less early anxiety and better postdischarge psychosocial adjustment compared with parents in the social support intervention at 2 wks after hospital discharge. Patterns of change over time, however, supported the efficacy of the CPR-social support intervention or the CPR-instructor protocols when compared with the CPR-video protocol. CONCLUSION: The results confirm that parents have difficulty adjusting after an infant's discharge from the neonatal intensive care unit and support the positive psychosocial effects of helping parents prepare for a home emergency by teaching CPR. The additional staff resources required to provide parents with social support along with CPR training are not justified based on the findings of the current study. PMID- 11008995 TI - Outcome of cardiopulmonary resuscitation in a pediatric cardiac intensive care unit. AB - OBJECTIVE: To determine the eventual outcome of children with heart disease who had cardiopulmonary resuscitation (CPR) in a specialized pediatric cardiac intensive care unit (CICU), and to define the influence of any prearrest variables on the outcome. DESIGN: A retrospective review of patients' medical records. SETTING: A pediatric CICU of a tertiary pediatric teaching hospital. PATIENTS AND METHODS: Patients were all children who presented with cardiopulmonary arrest and who were administered CPR in the pediatric CICU between June 1995 and June 1997. Prearrest variables such as age, diagnosis, prior cardiac surgery, and inotropic support with epinephrine, as well as cause of arrest, were evaluated. MEASUREMENTS AND MAIN RESULTS: Thirty-two patients, ranging in age from 1 day to 21 yrs (median, 1 month), satisfied criteria for inclusion in the study group. These 32 patients had a total of 38 episodes of cardiopulmonary arrest. Twenty-five of these patients (78%) had cardiac surgery before arrest. Inotropic support with continuous infusion of epinephrine was being administered at the time of arrest in 18 of 38 (47%) arrests. These prearrest variables did not influence outcome of CPR. Of the 38 episodes of CPR, 24 episodes (63%) were successful, with 20 episodes resulting in return of spontaneous circulation and four patients being successfully placed on mechanical cardiopulmonary support. Fourteen children, including all four patients who were rescued with mechanical cardiopulmonary support, survived to discharge. At 6 month follow-up, 11 patients were still alive, with three having neurologic impairment. CONCLUSIONS: After cardiopulmonary resuscitation in this pediatric CICU, the rate of success was 63% and the rate of survival was 42%. Prior cardiac surgery and use of epinephrine before arrest did not influence the outcome of CPR. The availability of effective mechanical cardiopulmonary support can improve the outcome of CPR. PMID- 11008996 TI - Use of hypertonic saline solutions in treatment of cerebral edema and intracranial hypertension. AB - OBJECTIVES: To review the literature on the use of hypertonic saline (HS) in treating cerebral edema and intracranial hypertension. DATA SOURCES: Review of scientific and clinical literature retrieved from a computerized MEDLINE search from January 1965 through November 1999. STUDY SELECTION: Pertinent literature is referenced, including clinical and laboratory investigations, to demonstrate principles and efficacy of treatment with HS in patients with intracranial space occupying pathology. DATA EXTRACTION: The literature was reviewed to summarize the mechanisms of action, efficacy, adverse effects, systemic effects, and comparisons with standard treatments in both clinical and laboratory settings. DATA SYNTHESIS: HS has an osmotic effect on the brain because of its high tonicity and ability to effectively remain outside the bloodbrain barrier. Numerous animal studies have suggested that fluid resuscitation with HS bolus after hemorrhagic shock prevents the intracranial pressure (ICP) increase that follows resuscitation with standard fluids. There may be a minimal benefit in restoring cerebral blood flow, which is thought to be mitigated through local effects of HS on cerebral microvasculature. In animal models with cerebral injury, the maximum benefit is observed in animals with focal injury associated with vasogenic edema (cryogenic injury). The ICP reduction is seen for < or =2 hrs and may be maintained for longer periods by using a continuous infusion of HS. The ICP reduction is thought to be caused by a reduction in water content in areas of the brain with intact blood-brain barrier such as the nonlesioned hemisphere and cerebellum. Most comparisons with mannitol suggest almost equal efficacy in reducing ICP, but there is a suggestion that mannitol may have a longer duration of action. Human studies published to date reporting on the use of HS in treating cerebral edema and elevated ICP include case reports, case series, and small controlled trials. Results from studies directly comparing HS with standard treatment in regard to safety and efficacy are inconclusive. However, the low frequency of side effects and a definite reduction of ICP observed with use of HS in these studies are very promising. Systemic effects include transient volume expansion, natriuresis, hemodilution, immunomodulation, and improved pulmonary gas exchange. Adverse effects include electrolyte abnormalities, cardiac failure, bleeding diathesis, and phlebitis. Although unproven, a potential for central pontine myelinolysis and rebound intracranial hypertension exists with uncontrolled administration. CONCLUSIONS: HS demonstrates a favorable effect on both systemic hemodynamics and intracranial pressure in both laboratory and clinical settings. Preliminary evidence supports the need for controlled clinical trials evaluating its use as resuscitative fluid in brain-injured patients with hemorrhagic shock, as therapy for intracranial hypertension resistant to standard therapy, as firstline therapy for intracranial hypertension in certain intracranial pathologies, as small volume fluid resuscitation during spinal shock, and as maintenance intravenous fluid in neurocritical care units. PMID- 11008997 TI - Thirty years of clinical trials in acute respiratory distress syndrome. AB - OBJECTIVE: To systematically review clinical trials in acute respiratory distress syndrome (ARDS). DATA SOURCES: Computerized bibliographic search of published research and citation review of relevant articles. STUDY SELECTION: All clinical trials of therapies for ARDS were reviewed. Therapies that have been compared in prospective, randomized trials were the focus of this analysis. DATA EXTRACTION: Data on population, interventions, and outcomes were obtained by review. Studies were graded for quality of scientific evidence. MAIN RESULTS: Lung protective ventilator strategy is supported by improved outcome in a single large, prospective trial and a second smaller trial. Other therapies for ARDS, including noninvasive positive pressure ventilation, inverse ratio ventilation, fluid restriction, inhaled nitric oxide, almitrine, prostacyclin, liquid ventilation, surfactant, and immune-modulating therapies, cannot be recommended at this time. Results of small trials using corticosteroids in late ARDS support the need for confirmatory large clinical trials. CONCLUSIONS: Lung protective ventilator strategy is the first therapy found to improve outcome in ARDS. Trials of prone ventilation and fluid restriction in ARDS and corticosteroids in late ARDS support the need for large, prospective, randomized trials. PMID- 11008998 TI - Prevention of nosocomial bloodstream infections: effectiveness of antimicrobial impregnated and heparin-bonded central venous catheters. AB - OBJECTIVES: To examine the effectiveness of antimicrobial-impregnated and heparin bonded catheters relative to standard central venous catheters in lessening catheter-related bloodstream infections. DATA SOURCES: Articles were identified by computer-assisted searching. STUDY SELECTION: Studies were eliminated from further consideration if they did not contain original data relevant to lessening catheter-related bloodstream infections, were nonrandomized or uncontrolled, described subjects <17 yrs of age, or used animal subjects. DATA ABSTRACTION: From each eligible article, we abstracted the following: a) citation; b) type of control; c) study setting; d) type of experimental catheter; e) catheter-specific complications; f) total numbers of patients and catheters; g) number of experimental catheters used that resulted in a catheter-related bloodstream infection; h) number of control catheters used that resulted in a catheter related bloodstream infection; i) number of experimental catheters used without catheter-related bloodstream infections; and j) number of control catheters used without infections. We also recorded the duration of catheter use and the types of microbes cultured in association with the catheters and with catheter-related bloodstream infections. DATA SYNTHESIS: Eleven eligible studies were identified. Using meta-analysis, we showed that antimicrobial-impregnated and heparin-bonded central venous catheters significantly decreased catheter-related bloodstream infections by 2.32% (95% confidence interval, 1.04% to 3.61%). CONCLUSIONS: The modest additional cost for the use of these catheters relative to the considerable cost of treating even a single bloodstream infection makes their use cost-effective. PMID- 11008999 TI - How to respond to family demands for futile life support and cardiopulmonary resuscitation. PMID- 11009000 TI - Reactive hemophagocytic syndrome presenting as a component of multiple organ dysfunction syndrome. AB - OBJECTIVE: To report two cases of severe reactive hemophagocytic syndrome (RHS), to discuss their impact, and to present evidence that RHS may be a constitutive part of multiple organ dysfunction syndrome (MODS). DESIGN: Case-report. SETTING: Pediatric intensive care unit (PICU). PATIENTS: Two patients with RHS and MODS. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Case #1: A 3 yr-old boy with Mucha-Haberman syndrome (pityriasis lichenoides) was admitted to the PICU for septic shock, acute respiratory distress syndrome, capillary leak, acute renal failure, liver dysfunction, and RHS (pancytopenia and hemophagocytosis on bone marrow aspirate). The pancytopenia was severe (white blood cell count, 0.9 x 10(9)/L; hemoglobin, 59 g/L; platelets, 36 x 10(9)/L), required many transfusions, and resolved 2 months later. The patient needed mechanical ventilation for 6 wks. Length of stay in PICU was 2 months. Case #2: A previously healthy 4 yr-old girl was admitted to the PICU for respiratory failure. She developed acute respiratory distress syndrome, cardiomyopathy with complete atrioventricular block, shock, capillary leak, liver dysfunction, and RHS (pancytopenia and hemophagocytosis on bone marrow aspirate). The pancytopenia was severe (white blood cell count, 1.92 x 10(9)/L; hemoglobin, 65 g/L; platelets, 58 x 10(9)/L) and necessitated transfusional support. Serology for respiratory syncytial virus was positive. RHS duration was 20 days; the patient recovered completely. Length of mechanical ventilation was 16 days and length of stay in PICU was 3 wks. CONCLUSIONS: These cases show that RHS may be a significant cause of pancytopenia in the PICU. It needs to be recognized as a clinical entity because it can be reversible and nonneoplastic. RHS and MODS share some pathophysiologic elements and could be related to each other. PMID- 11009001 TI - Hemoperfusion is ineffectual in severe chloroquine poisoning. AB - OBJECTIVES: To study the toxicokinetics in severe chloroquine poisoning, and to evaluate the efficacy of hemoperfusion. DESIGN: Case report on one observation. SETTING: Medical intensive care unit (ICU) of the University Medical Center Utrecht, The Netherlands. PATIENT HISTORY: A previously healthy, 52-yr-old woman ingested 100 tablets containing 100 mg chloroquine base 1 hr before admission. At admission, she was drowsy, agitated, hypotensive, and in respiratory distress. Shortly thereafter, she was resuscitated from cardiac arrest. After hemodynamic and respiratory stabilization, the patient was transferred to the medical ICU. TOXICOKINETICS EVALUATION: During the course of her stay at the ICU, blood samples were taken for the determination of chloroquine and the metabolite desethylchloroquine concentration. Hemoperfusion was started 3.5 hrs after ingestion of the chloroquine tablets. MEASUREMENTS AND MAIN RESULTS: The following toxicokinetics data during this severe chloroquine poisoning were calculated: apparent volume of the central compartment 181 L, apparent volume of distribution 1137 L, half-life in the distribution phase 6.4 hrs, half-life in the elimination phase 392.8 hrs, and total body clearance 2.01 L/hour. The average extraction ratio during hemoperfusion was 0.07, 0.28, and 0.25, in plasma, erythrocytes and whole blood, respectively. The total amount of chloroquine removed by hemoperfusion was only 480 mg (5.3% of the amount ingested). Simulation of a hemoperfusion session over 5 hrs by using a column with an optimal extraction ratio of 1.0 would have removed 1,816 mg chloroquine, only 18.2% of the amount ingested. This limited contribution of hemoperfusion to the total clearance makes it ineffective. CONCLUSION: Hemoperfusion is not effective in severe chloroquine poisoning, even when started (relatively) early in the course of the intoxication. Toxicokinetic evaluation of a chloroquine poisoning should be based on the evaluation of plasma and whole blood concentrations. PMID- 11009002 TI - Acute respiratory distress syndrome and protein: gold or flash in the pan? PMID- 11009003 TI - Optimizing the utilization of Advance Directives. PMID- 11009004 TI - Surgical models of acute lung injury. PMID- 11009005 TI - Thoracoabdominal aortic aneurysm repair: a human model of ischemia/reperfusion induced cytokine-driven multiple organ dysfunction syndrome. PMID- 11009006 TI - Another piece of the pathobiological puzzle in the response to trauma. PMID- 11009007 TI - Treatment of secondary ischemic insults after traumatic brain injury. PMID- 11009008 TI - Mucosal blood flow: acting globally but we should be thinking locally. PMID- 11009009 TI - Hyperventilation in severe brain injury revisited. PMID- 11009010 TI - The well-stressed intensivist. PMID- 11009011 TI - Are educational interventions enough for retention of cardiopulmonary resuscitation techniques? PMID- 11009012 TI - Cardiopulmonary resuscitation outcomes in children. PMID- 11009013 TI - Antimicrobial catheters: value and safety. PMID- 11009014 TI - Prediction of postoperative cardiac surgical morbidity and organ failure at admission to the intensive care unit using esophageal Doppler ultrasonography. PMID- 11009015 TI - Is surgical tracheostomy really superior to percutaneous tracheostomy? PMID- 11009016 TI - Percutaneous versus open tracheostomy: is it a fair comparison? PMID- 11009017 TI - Bedside percutaneous versus open tracheostomy. PMID- 11009018 TI - Steroids in cardiopulmonary bypass. PMID- 11009019 TI - Monitoring of peripheral nerves and muscle function in patients with multiple organ dysfunction syndrome. PMID- 11009020 TI - Saline-based fluids can cause a significant acidosis that may be clinically relevant. PMID- 11009021 TI - Dilutional acidosis. PMID- 11009022 TI - Venous air embolism from central venous catheterization: under-recognized or over diagnosed? PMID- 11009023 TI - Breath of life and cardiopulmonary resuscitation. PMID- 11009024 TI - Liquid adsorption chromatography of polyethers: experiments and simulation. AB - The adsorption behavior of poly(ethylene glycol) (PEG) in reversed-phase chromatography is studied both experimentally and theoretically and a computer simulation of chromatograms is performed on the basis of these studies. The experimental conditions were: different reversed-phase adsorbents and a solvent methanol-water system as the mobile phase. At varying mobile phase compositions highly resolved chromatograms of PEG samples were obtained, in which all peaks could be identified, and the dependencies of the distribution coefficient on the degree of polymerization for PEG molecules were evaluated by processing these chromatograms. The data were interpreted by using a theory of homopolymers based on a continuum Gaussian chain model of flexible macromolecules and a slit-like model of pores of stationary phase. The theory proved to describe well the experimental data in the whole range of studied molecular masses, and the thermodynamic parameters characterizing interactions of ethylene oxide repeating units in PEG molecules with the adsorbent pore walls have been determined from the comparison of the theory with the experimental data. The dispersion of chromatographic peaks corresponding to individual oligomer molecules is also estimated. In the system studied the peak width occurred to be proportional to the distribution coefficient of corresponding macromolecule. The theory is used to develop a computer-assisted procedure for simulation of chromatograms for samples of linear homopolymers. Using the obtained data on the thermodynamic parameters and the estimates of peak dispersion, chromatograms are simulated for PEG samples at two different chromatographic conditions. These simulated chromatograms were in good quantitative agreement with the real chromatograms. PMID- 11009025 TI - Determination of the lumped mass transfer rate coefficient by frontal analysis. AB - The validity of measurements of the lumped mass transfer rate coefficient (km,L) is studied on the basis of experimental data acquired under Langmuir isotherm conditions, in reversed-phase liquid chromatography. Two different methods were used, the perturbation method and frontal analysis. Accurate values of km,L can be properly obtained by the perturbation method because, with this method, the chromatographic processes take place under locally linear isotherm conditions. Values of km,L can also be derived from the breakthrough curves obtained in frontal analysis. Because the contribution of axial dispersion to band broadening was larger than that of the mass transfer resistances, the apparent axial dispersion coefficient (Da) was first derived from the breakthrough curve by applying the equilibrium-dispersive model. Then, the value of km,L was calculated from Da. The values of km,L determined by the two methods were in close agreement in the range of nondimensional Langmuir equilibrium constants (r= 1/[1+KLC0]) between 0.32 and 0.85, irrespective of the mobile phase flow velocity. Thus, frontal analysis can be used for kinetic studies of the mass transfer in chromatographic columns. PMID- 11009026 TI - Solid-phase extraction of polar hydrophilic aromatic sulfonates followed by capillary zone electrophoresis-UV absorbance detection and ion-pair liquid chromatography-diode array UV detection and electrospray mass spectrometry. AB - A comprehensive comparison of four different polymeric solid-phase extraction (SPE) materials for the extraction of 22 different aromatic sulfonates of environmental concern was performed. The investigated adsorbents were the polystyrene-divinylbenzene materials LiChrolut EN from Merck, Isolute ENV+ from International Sorbent Technology, HR-P from Macherey-Nagel and the new Oasis HLB poly(divinylbenzene-co-N-vinylpyrrolidone) copolymer from Waters. Different SPE parameters like the elution solvent and the drying step of the cartridges were optimized. Analyses were performed by capillary zone electrophoresis-UV absorbance detection (CZE-UV) and ion-pair liquid chromatography-diode array UV detection coupled in series with electrospray mass spectrometry (IP-LC-DAD-ESI MS) in the negative ionization mode. LC-MS offers a higher separation efficiency than CZE. The best adsorbents were LiChrolut EN and HR-P followed by Isolute ENV+ and Oasis HLB. The recoveries for most of the onefold negatively charged aromatic sulfonates were >50% for the extraction from spiked ground water at 50 microg/l. Recoveries for LiChrolut EN and HR-P were approximately 20% higher than for Isolute ENV+. Very hydrophilic sulfonates containing more than one negative sulfonate group could not be extracted by any of the tested adsorbents. PMID- 11009027 TI - Determination of the enantiomers of 3-tert.-butylamino-1,2-propanediol by high performance liquid chromatography coupled to evaporative light scattering detection. AB - A method for the separation and quantitation of the enantiomers of 3-tert. butylamino-1,2-propanediol by high-performance liquid chromatography and evaporative light scattering detection has been developed. Separation of the enantiomers was performed in normal-phase liquid chromatography on a Chiralpak AS chiral stationary phase. The influence of the gas nature, gas pressure and temperature of the drift tube of the evaporative light scattering detector on the detection sensitivity was investigated. The method was validated in terms of linearity, limit of quantitation, accuracy and precision. The enantiomeric excess of (S)-3-tert.-butylamino-1,2-propanediol, used for the industrial synthesis of (S)-timolol, was measured from 0 to 94%. PMID- 11009028 TI - Fractionation of apple procyanidins according to their degree of polymerization by normal-phase high-performance liquid chromatography. AB - A new method was developed for the fractionation of procyanidin oligomers according to their degree of polymerization. Monomeric flavan-3-ols and low molecular mass procyanidins were selectively extracted from the lyophilized powder of apple condensed tannins (ACTs) by methyl acetate extraction. Sequentially, the separation of each oligomer from dimer to pentamer in this extract was carried out by normal-phase high-performance liquid chromatography using a silica-beads packed column. The best separation was achieved with a mobile phase system containing hexane; (1) hexane-methanol-ethyl acetate, (2) hexane-acetone. These sequential treatments can be easily adapted to large-scale fractionation. PMID- 11009029 TI - Reinforcement of frontal affinity chromatography for effective analysis of lectin oligosaccharide interactions. AB - Frontal affinity chromatography is a method for quantitative analysis of biomolecular interactions. We reinforced it by incorporating various merits of a contemporary liquid chromatography system. As a model study, the interaction between an immobilized Caenorhabditis elegans galectin (LEC-6) and fluorescently labeled oligosaccharides (pyridylaminated sugars) was analyzed. LEC-6 was coupled to N-hydroxysuccinimide-activated Sepharose 4 Fast Flow (100 microm diameter), and packed into a miniature column (e.g., 10 x 4.0 mm, 0.126 ml). Twelve pyridylaminated oligosaccharides were applied to the column through a 2-ml sample loop, and their elution patterns were monitored by fluorescence. The volume of the elution front (V) determined graphically for each sample was compared with that obtained in the presence of an excess amount of hapten saccharide, lactose (V0); and the dissociation constant, Kd, was calculated according to the literature [K. Kasai, Y. Oda, M. Nishikawa, S. Ishii, J. Chromatogr. 376 (1986) 33]. This system also proved to be useful for an inverse confirmation; that is, application of galectins to an immobilized glycan column (in the present case, asialofetuin was immobilized on Sepharose 4 Fast Flow), and the elution profiles were monitored by fluorescence based on tryptophan. The relative affinity of various galectins for asialofetuin could be easily compared in terms of the extent of retardation. The newly constructed system proved to be extremely versatile. It enabled rapid (analysis time 12 min/cycle) and sensitive (20 nM for pyridylaminated derivatives, and 1 microg/ml for protein) analyses of lectin carbohydrate interactions. It should become a powerful tool for elucidation of biomolecular interactions, in particular for functional analysis of a large number of proteins that should be the essential issues of post-genome projects. PMID- 11009031 TI - Post-column oxidative derivatization for the liquid chromatographic determination of phenothiazines. AB - A first post-column chemical derivatization method for the liquid chromatographic determination of phenothiazines is presented. Peroxyacetic acid is introduced as a derivatizing agent for phenothiazines, yielding the colored radical cations or fluorescent sulfoxides, depending on reaction conditions. Both reaction products were successfully employed for the detection of the phenothiazines after their liquid chromatographic separation. The fluorescence spectroscopic detection of the sulfoxides proved to be the more robust and sensitive method. Limits of detection ranged from 4 nM for triflupromazine and trimeprazine to 300 nM for phenothiazine for the fluorescence spectroscopic detection of the sulfoxide and from 0.3 microM for phenothiazine and triflupromazine to 2 microM for trifluperazine for the UV-Vis spectroscopic detection of the radical cation. The calibration functions for the fluorimetric sulfoxide determination ranged from two to more than three decades, starting at the limit of quantification. PMID- 11009030 TI - Validation of an analysis method for 4-amino-3-hydroxybutyric acid by reversed phase liquid chromatography. AB - A rapid and simple reversed-phase liquid chromatographic method that did not require the derivatization of 4-amino-3-hydroxybutyric acid (GABOB) was developed and validated. The method proved to be suitable for the determination of GABOB concentrations in finished pharmaceutical product (tablets). The method was developed using a RP-18 column, UV detection at 210 nm and 0.01 M sodium heptasulphonate solution, at pH 2.4, as the mobile phase. Different validation parameters were included and satisfactorily evaluated. The specificity of the method was demonstrated. Linearity was established in the range 0.40-0.60 mg/ml (r=0.997). The method showed excellent accuracy (100.1%). Precision (repeatability) gave a relative standard deviation value of 0.68%, while the intermediate precision was 1.70%. A robustness test showing the influence of different pH values and counter-ion concentrations was also performed. PMID- 11009032 TI - Determination of naphthalenesulfonates in water by on-line ion-pair solid-phase extraction and ion-pair liquid chromatography with fast-scanning fluorescence detection. AB - A fast analytical method for quantifying a mixture of 12 naphthalenesulfonates and naphthalenedisulfonates has been developed. This method consists of on-line ion-pair solid-phase extraction with PLRP-s sorbent and ion-pair liquid chromatography using fast-scanning fluorescence spectrometer as a detection system and multivariate calibration. As complete separation is unnecessary, the compounds were analysed in isocratic conditions and the chromatographic analysis took only 25 min. Three-way partial least-squares (PLS) was used to carry out multivariate calibration for spiked tap water. In these conditions, quantification limits were between 0.01 and 3 microg x l(-1). Repeatability was also evaluated and relative standard deviations (n=3) were between 0.5 and 4, depending on the compound. Finally, spiked tap and Ebro river waters were analysed to evaluate prediction capability of the method. PMID- 11009033 TI - Simultaneous determination of inorganic disinfection by-products and the seven standard anions by ion chromatography. AB - For the first time, an ion chromatographic method for the simultaneous determination of the disinfection by-products bromate, chlorite, chlorate, and the so-called seven standard anions, fluoride, chloride, nitrite, sulfate, bromide, nitrate and orthophosphate is presented. The separation of the ten anions was carried out using a laboratory-made high-capacity anion-exchanger. The high capacity anion-exchanger allowed the direct injection of large sample volumes without any sample pretreatment, even in the case of hard water samples. For quantification of fluoride, chloride, nitrite, sulfate, bromide, nitrate, orthophosphate and chlorate, a conductivity detection method was applied after chemical suppression. The post-column reaction, based on chlorpromazine, was optimized for the determination of chlorite and bromate. The method detection limit for bromate measured in deionized water is 100 ng/l and for chlorite, it is 700 ng/l. In hard drinking water, the method's detection limits are 700 ng/l (bromate) and 3.5 microg/l (chlorite). The method's detection limits for the other eight anions, determined by conductivity detection, are between 100 microg/l (nitrite) and 1.6 mg/l (chlorate). PMID- 11009034 TI - Indirect analysis of urea herbicides from environmental water using solid-phase microextraction. AB - We described here a solid-phase microextraction procedure used to extract six urea pesticides-- chlorsulfuron, fluometuron, isoproturon, linuron, metobromuron and monuron--from environmental samples. Two polydimethylsiloxanes and a polyacrylate fiber (PA) are compared. The extraction time, pH control, addition of NaCl to the water and the influence of organic matter such as humic acid on extraction efficiency were examined to achieve a sensitive method. Determination was carried out by gas chromatography with nitrogen-phosphorus detection. The proposed method requires the extraction of 2 ml of sample (pH 4, 14.3%, w/v, NaCl) for 60 min with the PA fiber. The limits of detection range from 0.04 for linuron to 0.1 microg/l for fluometuron and monuron and the relative standard deviations at the 1 microg/l level are between 15% and 9%. The apparent fiber water distribution constants (Kfw) calculated in the proposed conditions were in the order of 10(3). Phenylurea herbicides were indirectly determined in the form of their derived anilines and chlorsulfuron in the form of an aminotriazine as confirmed by gas chromatography-mass spectrometry. Natural waters were utilized to validate the final procedure. However, a unequivocal identification in unknown environmental samples should be done by LC-MS. The presence of dissolved organic matter such as humic acid produces losses during the extraction step. Adding sodium chloride to the sample compensates for this effect. PMID- 11009035 TI - Retention index database for identification of general green leaf volatiles in plants by coupled capillary gas chromatography-mass spectrometry. AB - A series of ubiquitously occurring saturated and monounsaturated six-carbon aldehydes, alcohols and esters thereof is summarised as 'green leaf volatiles' (GLVs). The present study gives a comprehensive data collection of retention indices of 35 GLVs on commonly used non-polar DB-5, mid-polar DB-1701, and polar DB-Wax stationary phases. Seventeen commercially not available compounds were synthesised. Thus, the present study allows reliable identification of most known GLV in natural plant volatile samples. Applications revealed the presence of several seldom reported GLVs in headspace samples of mechanically damaged plant leaves of Carpinus betulus and Fagus sylvatica. PMID- 11009036 TI - Simulation of electrophoretic separations by the flux-corrected transport method. AB - Electrophoretic separations at typical experimental electric field strengths have been simulated by applying the flux-corrected transport (FCT) finite difference method to the transient, one-dimensional electrophoresis model. The performance of FCT on simulations of zone electrophoresis (ZE), isotachophoresis (ITP), and isoelectric focusing (IEF) has been evaluated. An FCT algorithm, with a three point, central spatial discretization, yields numerical solutions without numerical oscillations or spurious peaks, which have plagued previously-published second-order solutions to benchmark ZE and ITP problems. Moreover, the FCT technique captures sharp zone boundaries and IEF peaks more accurately than previously-published, first-order upwind schemes. PMID- 11009037 TI - Pressurized-flow anion-exchange capillary electrochromatography using a polymeric ion-exchange stationary phase. AB - The feasibility of using capillary columns equipped with silica frits and packed with a polymer-based anion exchanger (Dionex AS9-HC) for CEC separations of inorganic anions has been investigated. Experiments using a conventional 25 cm packed bed, and mobile phase flow that is a combination of hydrodynamic and electroosmotic flow were used to demonstrate that by varying the applied voltage (electrophoresis component) or the concentration of the competing ion in the mobile phase (ion-exchange component), considerable changes in the separation selectivity could be obtained. Using an artificial neural network, this separation system was modelled and the results obtained used to determine the optimum conditions (9 mM perchlorate and--10 kV) for the separation of eight inorganic anions. When a short (8 cm) packed bed was used, with detection immediately following the packed section, the separation of eight test analytes in under 2.2 min was possible using pressure-driven flow and a simple step voltage gradient. A more rapid separation of these analytes was obtained by only applying high voltage (-30 kV), where many of the same analytes were separated in less than 20 s and with a different separation selectivity to that obtained in conventional ion-exchange or capillary electrophoresis separations. PMID- 11009038 TI - Nonaqueous electrochromatography on continuous bed columns of sol-gel bonded large-pore C18 material: separation of retinyl esters. AB - A nonaqueous electrochromatographic reversed-phase separation method for retinyl esters using continuous bed columns has been developed. The packing material 7 microm Nucleosil 4,000 angstroms C18 was sol-gel bonded in 180 microm I.D. capillaries. The mobile phase used was 2.5 mM lithium acetate in N,N dimethylformamide-acetonitrile-methanol (2+7+1, v/v). At 350 V/cm and 30 degrees C, this mobile phase composition gave rise to an electroosmotic flow of 1 mm/s. No Joule heating nor bubble formation were observed even at 625 V/cm (17 microA). With a 36 cm L(eff) column complete separation of the commercially available and synthesized standards (all-trans-retinyl acetate, palmitate, heptadecanoate, stearate, oleoate, and linoleoate) was obtained within 10 min. The within-day and between-day variations of retention times of all-trans-retinyl palmitate were <0.3% relative standard deviation (RSD) (n=3) and <2% RSD (n=6), respectively. The within-day and between-day variations of peak areas were both <2% (both n=3). The columns were used for more than 1 month without degradation. Liver extracts from arctic seal were analyzed. PMID- 11009039 TI - New preconditioning strategy for the determination of inorganic anions with capillary zone electrophoresis using indirect UV detection. AB - It is widely accepted that preconditioning procedures are indispensable in capillary electrophoresis in order to achieve reproducibility of migration times and peak areas. Several preconditioning strategies have been employed for electrophoretic determinations of inorganic anions using indirect UV detection including simple flushing with buffer or alkaline or acid pre-rinsing followed by flushing with electrolyte. We investigated the influence of various preconditioning strategies on the reproducibility of migration times and peak areas of inorganic anions. The electrolyte systems for indirect UV detection were based on pyromellitic acid and chromic acid respectively as UV absorbing probes and hexamethonium hydroxide as electroosmatic flow modifier. Preconditioning agents under investigation were electrolyte buffer, NaOH, HCl and the free acids of the UV absorbing probes. Investigations showed that reproducibility of migration times and peak areas can be significantly improved by acid pre-rinsing using the corresponding acid of the UV absorbing probes compared to preconditioning by flushing the capillary with buffer. In contrast to acid pre rinsing using hydrochloric acid no interfering signals within the migration time window of inorganic anions under investigation can be observed. The optimized preconditioning procedure yields relative standard deviations of migration times less than 0.25% (n = 10). Relative standard deviations of corrected peak areas were below 5% applying acid preconditioning using pyromellitic acid. PMID- 11009040 TI - Potential of ethylenediaminedi(o-hydroxyphenylacetic acid) and N,N' bis(hydroxybenzyl)ethylenediamine-N,N'-diacetic acid for the determination of metal ions by capillary electrophoresis. AB - Two aromatic polyaminocarboxylate ligands, ethylenediaminedi(o hydroxyphenylacetic acid) (EDDHA) and N,N'-bis(hydroxybenzyl)ethylenediamine-N,N' diacetic acid (HBED), were applied for the separation of transition and heavy metal ions by the ion-exchange variant of electrokinetic chromatography. EDDHA structure contains two chiral carbon centers. It makes it impossible to use the commercially available ligand. All the studied metal ions showed two peaks, which correspond to meso and rac forms of the ligand. The separation of metal-HBED chelates was performed using poly(diallyldimethylammonium) polycations in mixed acetate-hydroxide form. Simultaneous separation of nine single- and nine double charged HBED chelates, including In(III), Ga(III), Co(II)-(III) and Mn(II)-(III) pairs demonstrated the efficiency of 40,000-400,000 theoretical plates. The separation of Co(III), Fe(III) complexes with different arrangements of donor groups and oxidation of Co(II), Mn(H), Fe(II) ions in reaction with HBED have been discussed. PMID- 11009041 TI - Use of voltage gradient gel electrophoresis in apoptotic DNA analysis. AB - In this paper the use of voltage gradient gel electrophoresis (VGGE) in the electrophoretic analysis of apoptotic DNAs is described. The peculiarity of VGGE fractionation in enhancing DNA bands in the gel by reducing their thickness was used to obtain a rapid, more selective and higher-quality electrophoretic fractionation of apoptotic DNA with respect to conventional electrophoresis. The use of VGGE fractionations also allowed a reduced amount of DNA to be used to detect a characteristic apoptotic DNA ladder pattern, in a lower agarose gel concentration, with respect to conventional electrophoretic fractionation PMID- 11009042 TI - The role of aminosalicylates at the beginning of the new millennium in the treatment of chronic inflammatory bowel disease. AB - OBJECTIVE: To discuss the pharmacological properties of aminosalicylates and their potential value in the treatment of chronic inflammatory bowel disease (IBD). METHODS: A review of clinical studies on the pharmacokinetics and mode of action of aminosalicylates is provided. In addition, the clinical efficacy and safety of aminosalicylates in the treatment of IBD, according to several recent meta-analyses, is summarised. RESULTS: Whereas aminosalicylates represent drugs of first choice in the acute treatment of ulcerative colitis and also for maintaining those patients in remission, their value for patients with Crohn's disease, either for achieving or maintaining remission, is at best modest. There is a large variability in the clinical results, especially in Crohn's disease, which is probably due to the variable extent and severity of IBD, different instruments in the evaluation of therapeutic outcome, and also at least partly caused by the different preparations and dosages of aminosalicylates used, as well as the high variation in drug disposition and topical availability of the active drug. The popular use of aminosalicylates is most likely due to the low incidence of side effects and the good overall safety records of mesalazine (mesalamine). CONCLUSIONS: Apparently, the full therapeutic potential of aminosalicylates has not yet been evaluated (e.g. upper dosage range, combination therapy, responding subgroups). Consequently, the imperfect treatment might be improved in the new millennium by novel insights in the complex mode of action of mesalazine as well as in the etiopathogenesis of IBD. PMID- 11009044 TI - Evaluation of the readability of information sheets for healthy volunteers in phase-I trials. AB - OBJECTIVE: The aim of the present study was to assess whether information sheets/consent forms submitted to the healthy volunteers of the Clinical Pharmacology Unit (C.P.U.) panel at Glaxo-Wellcome (Verona, Italy) could be considered understandable and to verify the readability and comprehensibility of these documents. Since a volunteer bases his/her decision to take part in a study on the information sheet provided, it is of paramount ethical importance to know whether the sheet conveys all relevant information. In addition, a thorough awareness by the volunteer of the reasons and procedures of the study would increase compliance. METHODS: Four indices were used: Flesh-Vacca, Kincaid, Gunning's Fog and Gulpease. All indices rate the degree of difficulty of a text, in the light of the level of schooling of the target population. The documents evaluated were information sheets presented to volunteers. The level of schooling of the population that participated in at least one study was determined: 61.7% of volunteers finished high school and 22.6% had a University degree or diploma; the remaining 15.7% did not finish high school or the datum was not available. RESULTS: The results showed that, when the present study began, all information sheets were "readable" by all volunteers who had at least finished high school. After these preliminary results, some additional linguistic and graphic refinements were adopted in drawing up information sheets. Readability improved to such a degree that all information sheets could be understood by virtually all volunteers. CONCLUSION: A number of suggestions were identified, which are set out in this paper to assist in the preparation of improved information sheets and a recommendation to value the readability of consent sheets before giving them to the volunteers. The suggestions were split into three categories: communications to the volunteer, text format and text organisation. PMID- 11009043 TI - Relative systemic dose potency and tolerability of inhaled formoterol and salbutamol in healthy subjects and asthmatics. AB - OBJECTIVES: To compare the relative systemic dose potency and tolerability of inhaled formoterol and salbutamol and to describe elimination of formoterol, particularly any enantioselectivity. METHODS: Twelve healthy subjects, aged 18-28 years, completed three open study days, and eleven asthmatic patients, aged 20-56 years, completed four double-blind study days in randomised, placebo-controlled and crossover fashions. The healthy subjects inhaled 13.5 + 13.5 + 27 microg formoterol (Oxis) via Turbuhaler and 300 + 300 + 600 microg salbutamol (Ventoline) via a pressurised metered dose inhaler (pMDI). The asthmatics, being on formoterol 9 microg twice daily via Turbuhaler during the study, inhaled the same single doses as the healthy subjects plus 900 + 900 + 1800 microg salbutamol via pMDI. Doses were given cumulatively 30 min apart on separate study days. Placebo was a day of no treatment in the healthy subjects. Double dummies were used for the asthmatics. Cardiovascular and metabolic effects were evaluated. Elimination of formoterol was addressed in the healthy subjects. RESULTS: Formoterol was estimated to be 28-109 times as potent as salbutamol, depending on the systemic effect variable. The duration of systemic action seemed to differ marginally at approximately equieffective doses of formoterol and salbutamol. Systemic effects were well tolerated and tended to be more pronounced in the healthy subjects than in the asthmatic patients. The half-life of the pharmacologically more active (R;R)-formoterol was longer than that of (S;S) formoterol. CONCLUSIONS: Systemically, formoterol was shown to be 28-109 times as potent as salbutamol. Equieffective doses seemed to have a similar duration of effect. Formoterol and salbutamol were well tolerated by healthy subjects up to the tested total doses of 54 microg and 1200 microg, respectively, and by asthmatic patients up to the tested total doses of 54 microg and 3600 microg, respectively. Elimination of formoterol was enantioselective. PMID- 11009045 TI - Cardiac effects of co-artemether (artemether/lumefantrine) and mefloquine given alone or in combination to healthy volunteers. AB - Co-artemether is an oral tablet of artemether (20 mg) and lumefantrine (120 mg) for the treatment of falciparum malaria. Administration in the presence of mefloquine is likely, as co-artemether may be used following failure of antimalarial prophylaxis or treatment with mefloquine. OBJECTIVE: The effects on the QTc interval were compared among treatment with three doses of mefloquine (500, 250, 250 mg over 12 h) followed by six doses of co-artemether (6 x 4 tablets over 60 h) and either treatment alone. The study was performed in a randomised, double-blind, parallel group design in 14 healthy male subjects per dose group. METHODS: Electrocardiograms (ECGs) were recorded before dosing and repeatedly thereafter. The Bazett formula was used to calculate the QTc interval. The maximum and average QTc intervals for the first, third and sixth dosing intervals of co-artemether treatment were compared among treatments. Drug plasma concentrations were determined at identical times with the ECG recordings for exploratory pharmacokinetic/pharmacodynamic evaluation. RESULTS: No clinically relevant differences in the QTc interval were observed after sequential administration of mefloquine and co-artemether relative to either treatment given alone, and there were no clinically relevant study drug-related effects on the QTc interval after either treatment. Plasma drug measurements revealed adequate systemic exposure to artemether, dihydroartemisinin, lumefantrine and mefloquine, well in line with the clinical setting. No correlation between the length of the QTc interval and plasma drug concentrations was found for any of the compounds. CONCLUSIONS: Untoward effects on the QTc interval are unlikely to occur when co artemether is administered following prophylaxis or treatment with mefloquine. PMID- 11009046 TI - A comparison of the effects of etodolac and ibuprofen on renal haemodynamics, tubular function, renin, vasopressin and urinary excretion of albumin and alpha glutathione-S-transferase in healthy subjects: a placebo-controlled cross-over study. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are known to be potentially nephrotoxic agents. NSAIDs inhibit the enzyme cyclo-oxygenase and thereby block the prostaglandin synthesis in the kidneys. Cyclo-oxygenase exists in two isoforms (COX-1 and COX-2). It has been proposed that NSAIDs with preferential COX-2 selectivity have fewer renal side effects than drugs with preferential COX-1 selectivity. Etodolac is a relative selective inhibitor of COX 2, while ibuprofen has a higher potency against COX-1 than COX-2. OBJECTIVE: In this study, we compared the effects of etodolac and ibuprofen on renal function, plasma renin, plasma arginine vasopressin and the urinary excretion of albumin and alpha-glutathione-S-transferase (alpha-GST). METHODS: In a randomised, double blind, three-way crossover study with placebo, we compared the effects of 2 weeks of treatment with ibuprofen and etodolac on renal haemodynamics [glomerular filtration rate (GFR), renal plasma flow (RPF) and filtration fraction (FF)], tubular function and plasma concentrations of the hormones renin (PRC) and arginine vasopressin (AVP) in 18 healthy subjects. In addition, we examined the effects on the urinary excretion of albumin and alpha-GST as markers of renal injury. RESULTS: No differences were found between the three treatments, placebo, ibuprofen and etodolac, in the effects on GFR, RPF, FF, free water clearance, urinary output or fractional excretion of potassium and sodium. However, ibuprofen, in contrast to etodolac, caused a significant decrease in both lithium clearance (-16% versus placebo) and the fractional excretion of lithium (-17% versus placebo), suggesting an increase in the reabsorption in the proximal tubuli. PRC was reduced significantly by ibuprofen (-32% versus placebo) but not etodolac. None of the drugs changed AVP. Fourteen days of treatment with ibuprofen caused a significant decrease (-47% versus placebo) in the urinary excretion of alpha-GST, while no changes were seen after etodolac. None of the drugs changed the urinary excretion of albumin. CONCLUSION: In conclusion, a 14 day administration of etodolac or ibuprofen in therapeutic doses did not affect the renal haemodynamics, the net excretion of electrolytes or the urinary excretion of albumin in healthy subjects. However, ibuprofen, in contrast to etodolac, caused a reduction in PRC, suggesting that COX-1 is involved in basal renin release in humans. Furthermore, ibuprofen decreased lithium excretion suggesting that COX-1 is involved in the re-absorption of sodium and/or water in the proximal tubuli. The reduction in the urinary excretion of alpha-GST by ibuprofen may be caused by an inhibition of the detoxification enzyme by ibuprofen. Overall the study indicates that only small differences in the effects of the two drugs on renal function in healthy subjects exist during a treatment period of 2 weeks. PMID- 11009047 TI - Concomitant use of mirtazapine and cimetidine: a drug-drug interaction study in healthy male subjects. AB - OBJECTIVE: The objective of this study was to examine the pharmacokinetics and the tolerability/safety of mirtazapine and cimetidine separately and in combination following oral administration of multiple doses. METHODS: This was a double-blind, placebo-controlled, two-period cross-over, multiple-dose pharmacokinetic interaction study in 12 healthy male subjects. They received either cimetidine (800 mg b.i.d.) or placebo in combination with (commercially available, racemic) mirtazapine (30 mg nocte). Cimetidine and placebo were administered for 14 days, with mirtazapine added during days 6-12 of each period. Serial blood samples for kinetic profiling were taken on day 5 and day 12 for cimetidine and on days 12-14 for mirtazapine. RESULTS: The co-administration of cimetidine resulted in a statistically significant increase in the area under the curve (AUC(0-24)) and Cmax of mirtazapine (54% and 22% respectively). The AUC(0 24) of demethylmirtazapine increased only slightly, and there was no effect on Cmax. The elimination half-lives for both mirtazapine and its demethyl metabolite were unaffected by cimetidine co-administration. The trough and average plasma concentrations during the steady state were elevated during cimetidine treatment (62% and 54%, respectively). Mirtazapine had no effect on the pharmacokinetics of cimetidine. CONCLUSION: Co-administration of cimetidine (800 mg b.i.d.) and mirtazapine (30 mg nocte) resulted in increased steady-state plasma levels of mirtazapine (C(ss,min) = +61%, P < 0.05; C(ss,av) = +54%, P < 0.05), probably as a result of increased bio-availability. The Cmax (+22%, P < 0.05) and AUC(0-24) (+54%, P < 0.05) also increased. Due to the variability of the mirtazapine plasma levels in patients, the clinical meaning of these increases is probably limited. Co-administration of mirtazapine did not alter cimetidine pharmacokinetics. PMID- 11009048 TI - Frequency of cytochrome P450 3A4 variant genotype in transplant population and lack of association with cyclosporin clearance. AB - OBJECTIVE: Cytochrome P450 3A4 (CYP3A4) plays a vital role in the oxidative metabolism of many xenobiotics. Some recent reports have provided circumstantial evidence in support of an association between a genetic polymorphism (A-->G) in the 5'-flanking region (-290) of CYP3A4 and altered enzyme activity. We sought to determine whether genotyping patients for CYP3A4-G could assist with the dose optimisation of drugs metabolised by this system. METHODS: Normal subjects and renal-transplant patients receiving cyclosporin for immune modulation were genotyped for the CYP3A4-G variant. A surrogate for cyclosporin clearance was estimated from the ratio of the cyclosporin dose, normalised for body weight and the corresponding trough concentration. The association between genotype and clearance was examined in patients who received twice-daily doses of cyclosporin and who were not on concurrent medication known to modify CYP3A4 function. RESULTS: The allelic frequencies of the CYP3A4-G variant were estimated to be 2.6% and 3% in transplant patients and normal subjects, respectively. The median cyclosporin pseudo-clearance of transplant patients with wild-type CYP3A4 was 0.90 l/h/kg (range: 0.35-3.8 l/h/kg; n = 86), whereas the corresponding value for the five patients heterozygotic for the CYP3A4-G variant was 0.71 l/h/kg (range 0.35-0.91 l/h/kg). The distribution of the pseudo-clearance according to genotype was not found to be significant according to a Fisher's exact test (P = 0.15). CONCLUSION: Genotyping for the CYP3A4-G polymorphism is unlikely to assist cyclosporin dose selection in transplant patients. PMID- 11009049 TI - Pharmacokinetics and pharmacodynamics of a premixed formulation of soluble and protamine-retarded insulin aspart. AB - OBJECTIVE: With the aim to obtain a premixed rapid-acting insulin with a serum insulin profile more closely resembling the endogenous meal-stimulated serum insulin profiles, a 30/70 (rapid/intermediate-acting) premixed suspension of the rapid-acting insulin analogue insulin aspart (BIAsp30) was compared with a similar premixed suspension of biphasic human insulin 30/70 (BHI30) after a single subcutaneous injection. METHODS: The study had a randomised, double-blind, two-period crossover design. Twenty-four healthy male subjects received a single subcutaneous dose of either 0.2 U x kg(-1) bodyweight of BIAsp30 or BHI30 on two study days. RESULTS: BIAsp30 was absorbed faster than BHI30, as reflected in the area under the insulin concentration-time curve from 0 to 90 min after dosing [AUC(0-90 min)]. This was significantly larger for BIAsp30 than for BHI30 (1403 +/- 372 versus 752 +/- 191 mU x l(-1) x min(-1) [mean +/- SD]; P < 0.0001). Furthermore, the time to maximum serum insulin concentration (tmax) of BIAsp30 was approximately half the tmax of BHI30 (60 [45-70] versus 110 [90-180] min [median, interquartile range]; P=0.0001) and the maximum insulin concentration (Cmax) was significantly higher for BIAsp30 than for BHI30 (23.4 +/- 5.3 versus 15.5 +/- 3.7 mU x l(-1) [mean +/- SD]; P < 0.0001). The serum glucose profiles showed a significantly earlier onset of the glucose-lowering effect following BIAsp30 than following BHI30. CONCLUSIONS: The improved absorption properties of soluble insulin aspart in its premixed formulation provide a basis for a more efficient meal-related glucose control and immediate pre-meal delivery when compared with a similar human premixed insulin in the treatment of diabetes mellitus. PMID- 11009050 TI - Metabolism of chloramphenicol succinate in human bone marrow. AB - OBJECTIVE/METHODS: The metabolism of chloramphenicol succinate (CAPS) by human bone marrow was studied in vitro using 75 marrow samples. Whole marrow samples were incubated with CAPS with or without reduced nicotinamide adenine dinucleotide phosphate for 1, 2 and 3 h at 37 degrees C. Ficoll-paque-separated marrow mononuclear cells and erythrocytes were similarly incubated. After precipitation and centrifugation, clear supernatant was analysed for the presence of metabolites using high-performance liquid chromatography. RESULTS: Only one metabolite was detected when CAPS was incubated for 3 h with whole marrow from 72 donors. Its retention time (RT 10.9 min) corresponded to chloramphenicol (CAP). When CAPS was incubated with samples of whole marrow, marrow mononuclear cells, marrow erythrocytes, marrow plasma and peripheral blood from one donor who had taken Traditional Chinese Medicine (TCM), three metabolite peaks were detected within 15 min to 1 h. The RT of two of these peaks corresponded to CAP and nitroso-CAP (RT 14.9 min), but one peak remained unidentified. These peaks were not detected in the control samples incubated without CAPS. Blood samples collected after 3 months and 6 months to reconfirm metabolic activity yielded no such metabolite peaks when incubated with CAPS for 1-3 h. Therefore induction of enzyme activity by TCM was suspected. Three metabolite peaks with the same RTs were also detected when CAPS was incubated for 3 h with whole marrow from two other donors. CONCLUSION: These studies demonstrated that CAPS may be metabolised to CAP and occasionally other metabolites in human bone marrow. This novel observation is particularly important because the bone marrow is known to be a target organ for chloramphenicol toxicity. PMID- 11009051 TI - Effect of grapefruit juice dose on grapefruit juice-triazolam interaction: repeated consumption prolongs triazolam half-life. AB - OBJECTIVE: Grapefruit juice inhibits CYP3A4-mediated metabolism of several drugs during first pass. In this study, the effect of grapefruit juice dose on the extent of grapefruit juice-triazolam interaction was investigated. METHODS: In a randomised, four-phase, crossover study, 12 healthy volunteers received 0.25 mg triazolam with water, with 200 ml normal-strength or double-strength grapefruit juice or, on the third day of multiple-dose [three times daily (t.i.d.)] administration of double-strength grapefruit juice. Timed blood samples were collected up to 23 h after dosing, and the effects of triazolam were measured with four psychomotor tests up to 10 h after dosing. RESULTS: The area under the plasma triazolam concentration time curve (AUC(0-infinity)) was increased by 53% (P < 0.01), 49% (P < 0.01) and 143% (P < 0.001) by a single dose of normal strength, a single dose of double-strength and multiple-dose administration of double-strength grapefruit juice, respectively. The peak plasma concentration (Cmax) of triazolam was increased by about 40% by a single dose of normal strength grapefruit juice (P < 0.01) and multiple-dose grapefruit juice (P < 0.01) and by 25% by a single dose of double-strength grapefruit juice (P < 0.05). The elimination half-life (t(1/2)) of triazolam was prolonged by 54% during the multiple-dose grapefruit juice phase (P < 0.001). A significant increase in the pharmacodynamic effects of triazolam was seen during the multiple-dose grapefruit juice phase in the digit symbol substitution test (DSST, P < 0.05), in subjective overall drug effect (P < 0.05) and in subjective drowsiness (P < 0.05). CONCLUSIONS: Even one glass of grapefruit juice increases plasma triazolam concentrations, but repeated consumption of grapefruit juice produces a significantly greater increase in triazolam concentrations than one glass of juice. The t(1/2) of triazolam is prolonged by repeated consumption of grapefruit juice, probably due to inhibition of hepatic CYP3A4 activity. PMID- 11009052 TI - Pattern of antibiotic use in primary health care in Italy. AB - OBJECTIVE: The overuse and misuse of antibiotics have been related to the growing emergence of bacterial resistance. The aim of the present study was to assess the pattern of antibiotic use by Italian general practitioners (GPs) in the treatment of the most frequent infectious problems. METHODS: The study was performed with 131 GPs recruited on a voluntary basis from among the 181 GPs contacted in two Italian regions, Emilia Romagna and Umbria. GPs were requested to report all the infectious events encountered during six sample weeks on a special form, whether an antibiotic was administered or not. RESULTS: The GPs reported 7095 infectious cases, of which 5036 (77%) were respiratory-tract infections (RTIs) and 749 (11%) were urinary-tract infections (UTIs). Antibiotics were prescribed in 71% of the cases. The proportion of antibiotic-treated cases was highest in UTIs (97%), followed by lower respiratory-tract infections (LRTIs; 93%) and upper respiratory tract infections (URTIs; 54%). Drugs belonging to 16 Anatomical Therapeutical Chemical groups (fourth level) were used. Wide-spectrum penicillins and macrolides ranked first (23%), followed by penicillins plus beta-lactamase inhibitors (15%), cephalosporins (15%) and fluoroquinolones (10%). The most prescribed antibiotics for the major disease groups were wide-spectrum penicillins for URTIs (36%), macrolides and cephalosporins for LRTIs (27% each) and fluoroquinolones for UTIs (46%). CONCLUSIONS: The present survey showed a high level of inappropriate use. In fact, a large number of infectious diseases, including infections commonly caused by viral agents, were treated with an anti bacterial drug. Italian GPs had a tendency to preferentially prescribe wide spectrum antibiotics and to use, in many cases, antibiotics that are rarely of choice in primary health care, such as cephalosporins and fluoroquinolones. In order to attain a more evidence-based prescription, local guidelines shared by specialists and GPs should be implemented. PMID- 11009053 TI - Incidence of lethal adverse drug reactions in the comprehensive hospital drug monitoring, a 20-year survey, 1974-1993, based on the data of Berne/St. Gallen. AB - OBJECTIVES: Realising the limitations of spontaneous drug monitoring systems concerning the epidemiological aspects, a comprehensive program was founded. It was based on previous publications from the US, Canada and Northern Ireland, mainly those of the BCDSP (Boston Collaborative Drug Surveillance Programme). METHODS: Drug monitoring was carried out by a group of physicians which included the medical head of each of the divisions of internal medicine, a statistician and an informatician. Only probable or definite drug event relationships were included. A probable event is defined as one in which the drug interaction was more likely to be the cause than any non-drug-related cause. The same criteria were valid for the lethal reactions. RESULTS: In the present evaluation, we found 26 probable lethal adverse drug reactions out of a total of 48,005 patients consecutively admitted to the divisions of internal medicine of three Swiss teaching hospitals during the years 1974-1993, an incidence of 0.054%. The median age of the cohort was 68 years (range 11-103 years), of which 49% were women. The median hospital stay was 14 days and the median number of drugs was eight per patient. CONCLUSION: The patients with a lethal outcome were presented under the eight pharmacologic-therapeutic classes of drugs and the classification proposed by NS Irey. This is based on long histopathologic experience and helps to identify preventable risks. PMID- 11009054 TI - Differences in the potencies of inhaled steroids are not reflected in the doses prescribed in primary care in New Zealand. AB - OBJECTIVE: To determine whether the average doses of inhaled beclomethasone, fluticasone and budesonide prescribed in primary care reflect the relative potencies of these medicines. METHODS: Retrospective analysis of 95,540 prescriptions for inhaled steroids written by 293 general practitioners in Auckland, New Zealand, between November 1995 and June 1998. In addition, 177 general practitioners were presented with two case histories describing patients with uncontrolled asthma who were not on treatment with inhaled steroids. They were asked which medicine they would prescribe and in what dose. RESULTS: The average daily doses prescribed were 600 microg for fluticasone, 747 microg for beclomethasone and 1184 microg for budesonide. The average dose of fluticasone was 80% of that for beclomethasone. In May 1997, when 4.5% of the prescriptions for inhaled steroids were for fluticasone, the average doses of fluticasone and beclomethasone were 632 microg and 760 microg, respectively. By May 1998, when 23% of prescriptions were for fluticasone, the average doses of fluticasone and beclomethasone were little changed at 610 microg and 726 microg, respectively. In response to the two case histories, the average doses of fluticasone chosen were 71% and 77% of the doses of beclomethasone. CONCLUSIONS: The average prescribed dose of fluticasone was 80% of that for beclomethasone, even though fluticasone is at least twice as potent as beclomethasone. Similar findings were observed when the general practitioners responded to the case histories. The high doses of fluticasone prescribed may be due to a failure to appreciate that fluticasone is twice as potent as beclomethasone and to the availability of high strength preparations of fluticasone, i.e. 250 microg per actuation. PMID- 11009055 TI - Do alcoholic beverages enhance availability of ivermectin? PMID- 11009056 TI - The effect of pantoprazole on tacrolimus and cyclosporin A blood concentration in transplant recipients. PMID- 11009057 TI - Policing the European pharmaceutical market's priorities. PMID- 11009058 TI - Effects of ethnicity on skeletal maturation: consequences for forensic age estimations. AB - An X-ray of the hand is an important method in forensic science for estimation of the age of juvenile suspects with uncertain date of birth. Relevant X-ray standards for evaluation of skeletal maturity are available for white US Americans as well as for North and Central Europeans. The applicability of these standards to members of ethnic groups different from the reference population has been the subject of controversial discussion. More than 80 publications were analysed with the view to finding out whether skeletal maturation is affected by ethnic identity. It was concluded that skeletal maturation takes place in phases which are identically defined for all ethnic groups. Time-related differences in passing those stages of skeletal maturation within the relevant age group appear to be unaffected by ethnic identity. It is the socioeconomic status of a given population which is of decisive importance to the rate of ossification. The application of X-ray standards to individuals of a socio-economic status lower than that of the reference population usually leads to underestimation of that person's age. In terms of criminal responsibility, this is of no adverse effect on the person concerned. PMID- 11009059 TI - Suicides by sharp force: typical and atypical features. AB - A total of 65 consecutive cases of suicide by sharp force were investigated by evaluating the autopsy and prosecution department records. Suicides constituted 17% of all fatalities from sharp force autopsied between 1967 and 1996. Young males and persons with a psychiatric history predominated among the persons who chose this "hard" method of suicide. The most common implements used were knives (62%) and razor blades (15%). Cutting injuries in isolation were present in 26, stab injuries in isolation in 24 and a combination of both in 15 fatalities. The number of injuries per case varied from 1 to 37 but 1/3 showed one injury. More than 85% of the cutting injuries were located at the wrist, elbow crease or neck whereas 79% of the stab injuries involved the ventral aspect of the trunk. Perforation of clothing was present in 16 (52%) out of 31 stab injuries to the trunk. Injuries to more than one body region were observed in 34 (52%) cases. Tentative marks were present in 50 (77%) fatalities and the number varied from 1 to 60 per case. Superficial incisions of the fingers were found in 15% with razor blades constituting the weapon in half of these cases. Deviations from these typical patterns occurred not infrequently. The utter determination of the victim to carry it through or the use of unusual weapons resulted in a few bizarre cases which are outlined briefly. PMID- 11009060 TI - Genetic variability at nine STR loci in the Chueta (Majorcan Jews) and the Balearic populations investigated by a single multiplex reaction. AB - A study of the genetic variability in the Chueta (Majorcan Jews) and the Balearic (Majorca and Minorca Islands) populations was carried out using a multiplex system containing the nine tetrameric STRs D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820. The Chueta population has remained isolated because intermarriage with non-Jews did not take place until the middle of this century, which has resulted in it being a small inbred community. The results indicate the existence of HW equilibrium for the Chueta and Balearic populations. No pair-wise correlation was observed between the nine markers. Consequently, they seem to comprise a suitable group of markers for population genetics purposes and for paternity and forensic testing. PMID- 11009062 TI - Analysis of eight STR loci in two Hungarian populations. AB - A multiplex reaction for the eight STR loci D3S1358, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820 was used to generate allele frequency databases for two Hungarian population samples, Caucasians from the Budapest area and Romanies from Baranya county. During the analysis two intermediate-sized alleles and a sequence variant allele were observed at the D7S820 locus. All three types of allelic variants were found to have modifications in the same block of a (T)9 stretch located within the 3' flanking region of each allele, which may indicate a possible higher mutation rate of this (T)9 block. For the loci D3S 1358 and D7S820 the Romany population database showed departures from Hardy-Weinberg equilibrium. The forensic efficiency values for the Romany population were slightly different from those found in the Hungarian Caucasian population. Comparing the allele frequency values by G-statistic, calculating the F(ST) indices and with the pair-wise comparisons of interpopulation variance, the two Hungarian populations could be distinguished using data from the eight STR loci. PMID- 11009061 TI - Immunohistochemical investigation of a pulmonary surfactant in fatal mechanical asphyxia. AB - We evaluated the usefulness of pulmonary surfactant protein A (SP-A) as a practical diagnostic marker of fatal mechanical asphyxia in forensic autopsy cases. A total of 27 cases of asphyxia were examined histologically and immunohistochemically and compared with a control group consisting of 16 cases of poisoning (n = 9) and peracute death (n = 7). Both groups showed histological findings of local atelectasis and local emphysema, congestion, intra-alveolar and interstitial edema in most cases and pulmonary hemorrhages in some cases. The mechanical asphyxia group showed a significantly increased intensity of SP-A staining in the intra-alveolar space accompanied by many massive aggregates in approximately 60% of cases, which was not found in the control group. These structures may be interpreted as aggregates of pulmonary surfactant released from the alveolar wall due to enhanced secretion caused by strong forced breathing or over-excitement of the autonomic nervous system by mechanical asphyxia. The results of our investigation suggest the practical usefulness of the immunohistochemical detection of SP-A in distinguishing mechanical asphyxia from other types of hypoxia. PMID- 11009063 TI - Myocardial findings in fatal carbon monoxide poisoning: a human and experimental morphometric study. AB - The aim of this study was to define the status of the myocardium in selected human cases of acute, fatal carbon monoxide intoxication and the myocardial changes in rats exposed to carbon monoxide in relation to the type of cardiac arrest and the effects of reoxygenation following pre-fatal CO intoxication. The human study consisted of 26 cases (17 accidental and 9 suicide) of acute, fatal CO intoxication, without evidence of obstructive coronary atherosclerosis or history of ischemic heart disease which were compared with 45 cases of fatal head trauma in subjects who died instantaneously (26 cases) or within 1-12 h (19 cases). Inhalation of a lethal dose of CO in rats was compared with sub-lethal doses plus reoxygenation with and without pre-treatment by a betablocker. In all human and experimental histological sections, changes were normalised per mm2 area. In the human cases the myocardium did not show any ischemic types of changes or other lesions. Only in "three accidental" cases a few, small foci of coagulative myocytolysis were detected. In the case of spontaneous death in 31 rats following CO intoxication, no pathological myocardial changes were seen. Of the 15 "reoxygenated" rats, 2 of the 7 spontaneous deaths presented coagulative myocytolysis with 15 +/- 6 foci and 381 +/- 255 necrotic myocells. All the eight rats sacrificed at 3 h had coagulative myocytolysis with 5 +/- 4 foci and 60 +/- 47 myocells. Of the 24 reoxygenated rats pre-treated with a betablocker, 5 died spontaneously after a short survival and 2 of these showed 11 +/- 9 foci and 21 +/- 20 myocells. The 19 rats sacrificed after 3 h all presented coagulative myocytolysis with figures of 75 +/- 43 and 356 +/- 301 with 0.5 mg/kg of propranolol hydrochloride and 55 +/- 45 and 253 +/- 216 with 2 mg/kg, respectively. PMID- 11009064 TI - Pressure sores: epidemiology, medico-legal implications and forensic argumentation concerning causality. AB - The aim of the present investigation was to identify the frequency and grading of pressure sores in a large series of unselected consecutive deceased subjects before cremation and to discuss aspects of the forensic argumentation concerning causality. A total of 10,222 corpses were examined prospectively over a 1-year period (1998) for the occurrence, localization and grading of pressure sores. Epidemiological aspects (e.g. age, sex, underlying and contributing causes of death, place of death etc.) were taken from the death certificate. The mean prevalence of pressure sores was 11.2% (87.1% isolated sores; predominant localization of advanced grades on the sacrum in 69.6%). There was a positive correlation between the prevalence of sores and advanced age resulting in a clear female predominance in the age group of 80 years and over because of differences in life expectancy. A significant correlation was found between the prevalence of pressure sores and certain underlying diseases, e.g. trauma, senile dementia, neurological diseases, apoplexy and marasmus. Pressure sores of all grades were more frequently found in the deceased when a senior citizen's or nursing home was given on the death certificate as the domicile in the last period of life. The forensic argumentation for the causal relationship of a pressure sore as the focus of fatal infectious complications or septicemia has to be based on the results of clinical expertises and forensic investigations (evaluation of the institutional documentation of the patient's course, autopsy findings, histology, immunohistochemistry). The vast majority of physicians seem to pay only little attention to the potentially fatal outcome of pressure sores and fatalities associated with this condition are clearly underreported. From the point of view of social medicine, the prevalence of pressure sores in a defined population can be seen as a parameter for the quality of nursing and medical care. In bringing these fatalities to light, the field of legal medicine contributes to a general quality control of standards of nursing and medical care. PMID- 11009065 TI - The time course of the vascular response to human brain injury--an immunohistochemical study. AB - In a total of 104 individuals who had sustained traumatic brain injury (TBI), the time-dependent vascular response was investigated at the injured cortical area during the first 30 weeks after the trauma. The immunohistochemical staining of the cerebral blood vessels was performed with antibodies against laminin, type IV collagen, tenascin, thrombomodulin and factor VIII associated antigen. Compared to the immunoreactivity in unaltered control tissue, a significantly increased vascular expression could be detected in cortical contusions after a postinfliction interval of at least 3 h for factor VIII, after 1.6 days for tenascin or after 6.8 days for thrombomodulin, whereas the immunostaining for laminin and type IV collagen was regularly positive even in the vascular endothelium of uninjured brain tissue. PMID- 11009067 TI - Polymarker, HLA-DQA1 and D1S80 allele data in a Zimbabwean Black sample population. AB - Allele frequencies for the seven PCR-based loci (LDLR, GYPA, HBGG, D7S8, Gc, HLA DQAI, and D1S80) were determined in a Black African population from Zimbabwe. All loci are highly polymorphic and meet Hardy-Weinberg expectations. An interclass correlation analysis detected only two significant departures from independence out of 21 pair-wise comparisons of the 7 loci. The Black African allele frequency data are similar to African American data at four of the seven PCR-based loci. PMID- 11009066 TI - ROC analysis of alcoholism markers--100% specificity. AB - A combination of 4 so-called markers of alcoholism, i.e. methanol, acetone + 2 propanol, gamma-glutamyltransferase and carbohydrate deficient transferrin, was investigated in 341 blood samples from alcoholics and non-alcoholics. From the history of alcohol consumption, four defined subgroups were formed: non alcoholics divided into (A) 33 persons with no ethanol consumption during the past year and (B) 60 persons with daily consumption less than 40 g ethanol. Alcoholics were divided into (C) 177 persons with no ethanol at the time of admission/first blood sampling (withdrawal therapy) and (D) 71 persons with positive ethanol levels on admission/first blood sampling. All markers showed different extents of overlap between the collectives of alcoholics and non alcoholics. By logistic regression, a formula was developed combining these markers with different mathematical weights. Thus an "Alc-Index" could be calculated for each individual. The ROC curve connecting all individual values gives an ideal form with 100% specificity and nearly 93% sensitivity. The threshold between the collectives of alcoholics and non-alcoholics was defined by the Alc-Index value 1.7. This was associated with no false positives among the non-alcoholics while nearly 93% of the alcoholics exceeded this index. The ROC based calculation of the Alc-Index thus seems to be the most effective method for the diagnosis of alcoholism. PMID- 11009069 TI - Frontal lobe membrane phospholipid metabolism and ventricle to brain ratio in schizophrenia: preliminary 31P-MRS and CT studies. AB - A number of studies employing in vivo phosphorous-31 magnetic resonance spectroscopy (31P-MRS) have demonstrated altered measurements of frontal phospholipid and high energy phosphorus metabolism in schizophrenia. Enlargement of both the cerebroventricular system and the cortical sulci also has been reported as the most consistent pathological finding in schizophrenia by several investigators. To our knowledge, however, only two studies have simultaneously examined structural and functional aspects of the biological substrate of schizophrenia in the same patients. Moreover, they may have failed to find a significant correlation between these variables because of small sample sizes. The possible relationship between frontal lobe membrane phospholipid metabolism and ventricle-to-brain ratio (VBR) in patients with schizophrenia was investigated. In 31 schizophrenic patients, frontal lobe membrane phospholipid metabolism was measured by 31P-MRS, and VBR was measured by computed tomography (CT). Stepwise multiple regression analysis disclosed that VBR positively correlated only with increased phosphodiester (PDE) level (beta = 0.381, p = 0.0345), but with no other metabolites. This finding may provide evidence for an association between structural brain abnormality and altered frontal lobe membrane metabolism in schizophrenic patients, although the p-value of the finding is not high. PMID- 11009068 TI - hKCNN3 which maps to chromosome 1q21 is not the causative gene in periodic catatonia, a familial subtype of schizophrenia. AB - The human calcium-activated potassium channel gene (hKCNN3, hSKCa3) contains two tandemly arranged, multiallelic CAG repeats located in exon 1 which result in short to moderate polyglutamine stretches of unknown functional significance. Case-control and family-based association studies suggested an association of hKCNN3 repeats with susceptibility for schizophrenia. Twelve multiplex pedigrees with periodic catatonia, a schizophrenia subtype with major gene effect and patterns of anticipation, were genotyped using the multiallelic hKCNN3 repeat polymorphism. Using a dominant model of inheritance with sex- and age-dependent penetrance classes, cumulative results showed exclusion of linkage of hKCNN3 to periodic catatonia under the assumption of genetic homogeneity with lod score of 48.01 at zero recombination fraction. Our results provide evidence that hKCNN3 is not the causative gene in the familial schizophrenia subtype of periodic catatonia. By fluorescent in situ hybridization we confirmed the assignment of hKCNN3 to chromosome 1q21 near the heterochromatin region. Linkage mapping showed segregation with marker D1S498 (theta = 0.05) and placed hKCNN3 in the genetic linkage map in a cluster of genes near the centromeric region of chromosome 1. PMID- 11009070 TI - Comparing self- and expert rating: a self-report screening version (SIAB-S) of the structured interview for anorexic and bulimic syndromes for DSM-IV and ICD-10 (SIAB-EX). AB - OBJECTIVE: Carrying out structured interviews in larger numbers by well-trained interviewers is costly and time consuming. Therefore, we developed parallel to the existing Structured Interview for Anorexic and Bulimic Syndromes (SIAB-EX) a similarily designed questionnaire for symptoms of disordered eating and related areas (SIAB-S). METHOD: 377 treated eating disordered patients were assessed within a two-week time period using both the SIAB-EX and SIAB-S. RESULTS: Generally, self-ratings based on the SIAB-S were quite similar to expert ratings. Cohen's kappa showed good agreement between self- and expert ratings. Factor structure based on principal component analyses of expert ratings or self-ratings led to rather similar results confirming the robustness of the subscales in self- and expert ratings. Using expert rating as a criterion, the self-rating (SIAB-S)- which can more easily be used for screening purposes--had a sensitivity of 0.70, a specificity of 0.80 and a PPV = 0.91 for the DSM-IV diagnoses of AN and/or BN (worst ever condition). Diagnostic sensitivity (79/73%) and specificity (66/63 %) were in an acceptable range (past/current). If we focus on the differences between the two approaches the following was found: self-rating (compared to expert-rating) resulted in lower scores for items inquiring about binges and inappropriate compensatory behaviour, attitudes towards food and eating, and social interaction. On the other hand, self-rating (compared to expert-rating) led to higher scores for items measuring general psychopathology and atypical binging. CONCLUSION: Compared to the "gold standard" of t data obtained with investigator-based standardised or structured interviews, data based on self rating with items formulated clearly and concisely can lead to reliable and valid results. While complex issues (what is a binge) are difficult to assess in self ratings, some (very personal) questions may even be better asked in a self-report questionnaire. PMID- 11009071 TI - Comorbidity of mild cognitive disorder and depression--a neuropsychological analysis. AB - Mild cognitive impairment is found in many cases of depression, and it is mostly assumed to improve during the time course of depression remission. Recent data question the reversibility of low cognitive test performance in depression. The aim of this study is to determine the degree of reversibility and the proportion of patients who will not demonstrate reversibility of cognitive dysfunction. Consecutive inpatients suffering from depression (N=102) were investigated and N=82 matched control subjects. N=57 of the patients were diagnosed as major depression according to DSM-IV. A total of N=67 could be retested after remission of depression (N=32 of the patients with major depression) and a matched control group (N=62). Neuropsychological tests were applied in a test session which avoids the effects of fatigue in the patients by the short duration of strenuous tests. For most neuropsychological tests an impaired performance in the depressed patients was found. About one third of the depression subjects performed at an impaired level in tests of averbal memory and verbal fluency (below 5th percentile). In the follow-up investigation, a slight improvement in performance could be assessed for both the depression and the control group, which was, however, attributed to a general test training effect. No normalization of cognitive test performance was found in spite of complete recovery of the affective symptoms. No correlation between the duration of the disease before the index episode or number of episodes and cognitive deficits could be found. The data of the neuropsychological deficits of depressed patients, which are stable in the time course of the affective disorder, may indicate that these patients may suffer from comorbidity of both depression and mild cognitive disorder. The findings are discussed as 1) indicating only a minor impact of the depressed mood on the cognitive performance and 2) they are consistent with a role of brain lesions which have been reported in several studies in a subgroup of depression. PMID- 11009072 TI - Naturally occurring benzodiazepines: current status of research and clinical implications. AB - Naturally occurring benzodiazepines (BZDs) were first detected in mammalian tissues in 1986. They comprise a variety of 1,4-benzodiazepines corresponding to drugs commercially available for the treatment of anxiety disorders, sleep disturbances and epileptic seizures. Several biosynthetic pathways leading to the formation of BZDs are currently being discussed and have led to the proposition of possible precursor molecules. For years, the identification of naturally occurring BZDs in mammalian organisms was mostly confined to post mortem CNS material for sensitivity reasons. While radioimmunoassay and radioreceptor assay techniques have been tentatively applied to quantitations of genuine BZDs from human milk and cerebrospinal fluid, accurate measurements in peripheral blood have only recently become accessible, e. g., by gas chromatography/selected ion monitoring-mass spectrometry (GC/SIM-MS). This review summarizes existing evidence of benzodiazepines' occurrence in nature and discusses implications for neuropsychiatric disorders. PMID- 11009073 TI - Dopamine D4 receptor gene polymorphism and personality traits in healthy volunteers. AB - An association between long alleles of a variable number tandem repeat (VNTR) polymorphism in the dopamine receptor D4 gene and the extraversion related personality traits Excitement and Novelty Seeking has been reported in healthy subjects. In an attempt to replicate the previous findings, 256 healthy Caucasian volunteers were analysed for a potential relationship between the dopamine receptor D4 exon III VNTR polymorphism and Extraversion as assessed by the Revised Neo Personality Inventory (NEO PI-R). The present study did not yield evidence for an association between Extraversion and the dopamine receptor D4 polymorphism. PMID- 11009074 TI - Frontal dysfunction in schizophrenia--a new electrophysiological classifier for research and clinical applications. AB - We determined whether schizophrenic patients can be reliably classified with electrophysiological tools. We developed a fully computerized classifier based on 5 minutes of EEG recording during an acoustical choice reaction time task (AMDP module IV). We included factorized variables from the frequency domain and evoked potentials (N100/P200-complex) from central and frontal electrodes, which were preprocessed in a sample of 150 normal subjects prior to classification. We applied discriminant analyses to the electrophysiological data from depressive, schizophrenic and schizotypal subjects, most of them being unmedicated or drug naive. The classifier was developed on a training set (33 schizophrenics, 49 normals) and tested on an independent sample (32 schizophrenics, 49 normals). A simple three-variable classifier was found to classify schizophrenics and normals in 77% of those tested correctly. Diagnostic specificity of the classifier proved to be low as the inclusion of depressive patients (n= 60) significantly decreased classification power. It was demonstrated that atypical but not typical neuroleptic drugs may influence the classification results. Correctly classified schizophrenics showed significantly more negative symptoms and slower reaction times than those schizophrenics who were misclassified as normals. In contrast, these misclassified schizophrenics showed a non-significant trend for more positive symptoms and shorter reaction times. As the correctly classified schizophrenics showed increased frontally pronounced delta-activity and decreased signal power of the N100/P200 amplitude, it was concluded that these schizophrenics show dysfunction of the frontal lobe. It is proposed that this new classifier can be useful for clinical and research applications when subtyping of schizophrenics with detection of frontal dysfunction as the aim. PMID- 11009075 TI - Mitogen-activated kinase kinase kinase 1 regulates T cell receptor- and CD28 mediated signaling events which lead to NF-kappaB activation. AB - Optimal activation of Rel/NF-kappaB transcription factors in T lymphocytes requires a CD28-delivered co-stimulatory signal in addition to TCR engagement. Although, Rel/NF-kappaB transcription factors are critical regulators of many T cell functions, the mechanisms and molecules, which link the surface receptors to their activation, are poorly characterized. Using Jurkat T cells stimulated with superantigen presented on B7-positive APC, we showed that CD28- and TCR stimulated NF-kappaB-dependent transcription is associated to the activation of IkappaB kinase beta (IKKbeta) and, to a lesser extent, of IkappaB kinase alpha (IKKalpha). A dominant negative mutant of the MAP3 kinase MEKK1, a kinase known to regulate the JNK pathway and to activate NF-kappaB-dependent transcription in many cell types, strongly inhibits CD28- and TCR-induced IKK activity, whereas the dominant negative mutants of the NF-kappaB-inducing kinase (NIK) did not exert any significant effects. In addition, TCR/CD28 stimulation results in the recruitment and autophosphorylation of endogenous MEKK1, whereas endogenous NIK was not detectably activated. Our data identify MEKK1 as a critical step in coupling signals initiated by TCR and CD28 to the downstream pathways which lead to both AP-1 and NF-kappaB activation in T lymphocytes. PMID- 11009076 TI - CD4+ T cells contain Mycobacterium tuberculosis infection in the absence of CD8+ T cells in mice vaccinated with DNA encoding Ag85A. AB - The contribution of CD8+ and CD4+ T cell-mediated effector functions against Mycobacterium tuberculosis infection elicited by i.m. vaccination with plasmid DNA encoding the immunodominant Ag85A antigen of M. tuberculosis was studied. Ag85A DNA-vaccinated beta2-microglobulin gene-deficient (beta2m-/-) mice, which lack CD8+ T cells, produced Ag85-specific antibodies and Th1 type cytokines similar to wild-type mice. Although beta2m-/- mice were more susceptible to M. tuberculosis infection, following vaccination they efficiently controlled bacterial replication in spleen and lungs 4 weeks post-infection. In contrast, mice lacking CD4+ T cells were neither sensitized by the Ag85A DNA vaccine to produce Ag85-specific antibodies or Th1 type cytokines nor did they contain a M. tuberculosis challenge infection. In addition, Ag85A DNA-vaccinated IFN-gamma gene knockout mice produced Ag85-specific antibodies and IL-2 but died rapidly following a M. tuberculosis challenge infection. Collectively, these data support the view that IFN-gamma-producing CD4+ T cells, independently of CD8+ T cells, may mediate the protective effect of the Ag85A DNA vaccine. PMID- 11009077 TI - Anti-TCR Vbeta-specific DNA vaccination prolongs heart allograft survival in adult rats. AB - Allospecific T cells are known to play a central role in the process of allograft rejection. Recently, it has been shown that T cell function could be specifically targeted using DNA vaccination. In our model, PCR analysis of the TCR-beta chain repertoire of T cells infiltrating rejected allografts showed specific expansions of the Vbeta13 and Vbeta2 families. In this study, we tested the effect on allograft survival of DNA vaccination against a specific TCR Vbeta, in a model of heart allograft rejection in adult rats. Our results showed that anti-TCR Vbeta13 DNA vaccination lead to a significant prolongation of allograft survival compared to vaccination against other Vbeta families or untreated recipients. The prolongation of allograft survival correlated in vitro with a decrease in anti donor reactivity of spleen cells from Vbeta13-vaccinated rats. These results show that, in a transplantation model, DNA vaccination could be used as a method to specifically manipulate a T cell response and thus prolong allograft survival. PMID- 11009078 TI - c-Myc inhibits CD11a and CD11c leukocyte integrin promoters. AB - The c-Myc transcription factor is an important regulator of cell growth and differentiation, and its gene repression ability seems to play a key role in Myc mediated cellular transformation. Since Myc overexpression has been associated with reduced expression of beta1 and beta2 integrins, we have investigated the role of c-Myc on CD11a and CD11c transcription. c-Myc inhibited CD11a and CD11c integrin promoter activity in co-transfection experiments, and similar repression was obtained in cells where c-Myc expression (KmycB) or activity (Rat-1 c-MycER) is inducible. The c-Myc repression on the CD11c promoter was independent of the USF-binding site (USF-150), other putative Myc-binding elements, or the integrity of the initiator (Inr)-like sequence present at the major transcriptional start site. Analysis of deletion and mutant promoter constructs revealed that, in the absence of additional upstream cisacting elements, an AP-1-binding site at -60 (AP1-60) is required for c-Myc repressor activity. The c-Myc repressor activity on both integrin promoters was abrogated by deletion of c-Myc residues 106-143, a domain involved in Inr-dependent transcriptional repression. These results demonstrate a direct effect of c-Myc on integrin gene transcription and suggest the existence of a c-Myc-dependent mechanism for coupling leukocyte integrin expression to the cell proliferative state. PMID- 11009079 TI - Developmental regulation of TCR efficiency. AB - Unresponsiveness of mature T cells to the same self-peptide/self-MHC molecule complexes with which thymocytes have reacted during positive selection results from an increase of activation thresholds during maturation. The molecular events accounting for this increase are still unknown. In mature cells, a strong correlation between the extent of TCR down-modulation and T cell responses has been demonstrated. Exploiting this relationship, we show that the efficiency with which the TCR mediates full signaling is developmentally regulated. It decreases with thymic maturation and then increases with differentiation of naive T cells into memory lymphocytes. This analysis based on TCR modulation revealed an unexpected characteristic of the T cell receptor: its ability to signal fully is regulated inversely by its level of expression. This latter relationship may apply to other receptors both within and outside the immune system. PMID- 11009080 TI - CD8+ T lymphocyte responses are induced during acute hepatitis C virus infection but are not sustained. AB - Cellular immune responses are likely to play a key role in determining the clinical outcome in acute infection with hepatitis C virus (HCV), but the dynamics of such responses and their relationship to viral clearance are poorly understood. In a previous study we have shown highly activated, multispecific cytotoxic T lymphocyte responses arising early and persisting in an individual who subsequently cleared the virus. In this study the HCV-specific CD8+ lymphocytes response has been similarly analyzed, using peptide-HLA class I tetramers, in a further nine individuals with documented acute HCV infection, six of whom failed to clear the virus. Significant populations of virus-specific CD8+ lymphocytes were detected at the peak of acute hepatic illness (maximally 3.5% of CD8+ lymphocytes). Frequencies were commonly lower than those seen previously and were generally not sustained. Early HCV-specific CD8+ lymphocytes showed an activated phenotype in all patients (CD38+ and HLA class II+), but this activation was short-lived. Failure to sustain sufficient numbers of activated virus-specific CD8+ lymphocytes may contribute to persistence of HCV. PMID- 11009081 TI - Differential regulation of Ly49 expression on CD4+ and CD4-CD8- (double negative) NK1.1+ T cells. AB - The pan-NK cell marker NK1.1, present in some mouse strains, is also found on a subset of TCRalphabeta+ lymphocytes termed NKT cells. These cells are primarily CD4+ or CD4-CD8- (double negative, DN), and both NKT subpopulations contain cells reactive with the MHC class I-like molecule CD1d. Murine NK cells express clonally distributed inhibitory receptors of the Ly49 family that bind to different alleles of MHC class I molecules and transmit negative signals regulating NK cell function. Ly49 receptors are also found on TCRalphabeta+ NK1.1+ T cells. To investigate the potential role of inhibitory Ly49 markers in the regulation of NKT cells, we have done a thorough analysis of their expression on different NKT populations. The CD4+ and DN NK1.1+ T cell subsets have traditionally been dealt with as one NK1.1+ T cell population, but we found dramatic differences between these two NKT cell subsets. We demonstrate here expression of Ly49 receptors on DN, but not on CD4+, NK1.1+ T cells in spleen and liver. Absence of the specific MHC class I ligand in the host was associated with elevated levels of expression and, to a greater extent than has been found for NK cells, increased the frequencies of Ly49-positive cells within the DN subset, while CD4+ NK1.1+ cells remained negative. In the thymus and bone marrow both NK1.1+ T cell subsets contained high frequencies of Ly49-positive cells. Results from in vitro stimulation of DN NKT cells further suggest that activation and expansion of NKT cell subsets are regulated by the Ly49 receptors. PMID- 11009082 TI - Binding of conserved islet peptides by human and murine MHC class II molecules associated with susceptibility to type I diabetes. AB - The major histocompatibility complex (MHC) is the most important susceptibility locus for type I diabetes in humans and NOD mice. NOD mice express a single MHC class II molecule (I-Ag7) which carries a unique beta chain sequence. In humans, DQ alleles that encode DQ8 and DQ2 confer the highest risk for the disease. Soluble DQ8 and I-Ag7 were used to directly compare the binding specificity of these MHC molecules. Peptides from three islet antigens--insulin, GAD 65 and HSP 60--bound to both CQ8 and I-Ag7. These peptides included epitopes that are immunodominant in NOD mice, namely insulin (9-23), GAD (206-220) and HSP 60 (441 460). All of these peptide sequences are highly conserved between the human and murine antigens. The binding specificity of DQ8 and I-Ag7 was similar, but not identical, since two peptides eluted from splenocytes of NOD mice did not bind to DQ8. DQ8 formed long-lived complexes with the majority of these peptides, indicating that DQ8 is not a poor peptide binder. These results demonstrate functional similarities between human and murine MHC class II molecules that confer susceptibility to type I diabetes. PMID- 11009083 TI - Differential requirement of perforin and IFN-gamma in CD8 T cell-mediated immune responses against B16.F10 melanoma cells expressing a viral antigen. AB - Perforin-mediated lysis and secretion of IFN-gamma belong to the key effector functions of CD8 T cells. To compare the anti-tumor activity of these two mechanisms, we used B16.F10 melanoma cells (B16GP33) expressing the cytotoxic T cell epitope GP33 and T cell receptor transgenic (TCR-tg) mice specific for GP33 and deficient in perforin or IFN-gamma. B16GP33 tumor cells, injected either i.v. to induce experimental metastases or s.c., were similarly controlled in both wild type and perforin-deficient, but not in IFN-gamma-deficient TCR-tg mice. A similar result was obtained when the therapeutic efficacy of adoptively transferred TCR-tg effector cells from these mice was examined in the corresponding perforin- or IFN-gamma-deficient C57BL/6 hosts. Criss-cross experiments further revealed that IFN-gamma production by the host strongly influenced the efficiency of the adoptively transferred effector cells. In contrast to the potent ability of GP33-specific effector cells to mediate B16GP33 tumor regression without perforin, GP33-specific memory cells, induced with recombinant vaccinia virus expressing GP33, failed to control B16GP33 tumor growth in the absence of perforin. In conclusion, our data demonstrate a crucial role for IFN-gamma in B16GP33 tumor cell elimination in vivo and indicate a differential requirement of perforin by effector versus memory CD8 T cells in anti-tumor immunity. PMID- 11009084 TI - Study of the B cell memory compartment in common variable immunodeficiency. AB - We evaluated the B cell memory pool among blood B cells from 20 patients with common variable immunodeficiency (CVID). CD27+ B cell number was normal or increased in 6 patients (with 95% CD27+ B cells in 1 patient) and decreased in 14 patients. In 13 or 15 patients studied, the CD27 molecule was detectable on less than 50% IgG or IgA B cells, indicating a defect in the maturation of these memory cells. Within the group of patients with a low number of CD27+ B cells, no up-regulation of this molecule was observed after in vitro stimulation of purified B cells from 3 of 5 patients studied, suggesting an intrinsic B cell defect. In addition, ligation of the CD27 molecule was unable to trigger terminal differentiation of purified B cells in 1 of 2 cases with a large number of CD27+ B cells. Finally, the CD27 ligand was normally expressed on activated T cells in only 5 of 14 patients studied. These data confirm the heterogeneity of immunological defects in patients with CVID. Abnormal expression and/or function of the CD27-CD70 members of the TNF/TNF receptor family contribute to the immunological defect. PMID- 11009085 TI - A single CTL clone can recognize a naturally processed HIV-1 epitope presented by two different HLA class I molecules. AB - Although it is known that a single peptide can be recognized by CTL restricted to two MHC class I alleles, there is no direct evidence for presentation of a single peptide by two MHC class I molecules. Furthermore, it is unclear whether such peptides are presented to the same T cell or to different T cells. Our previous study suggested that CTL recognition of the human immunodeficiency virus-1 (HIV 1) Pol HIV-B35-SF2-24 epitope (IPLTEEAEL) occurs via both HLA-B35 and HLA-B51 restriction. Here we provide the first direct evidence that a single CTL clone can recognize this peptide presented by both HLA-B35 and HLA-B51. Furthermore, we directly purified this peptide eluted from both HLA-B*3501 and HLA-B*5101 molecules isolated from target cells infected with HIV-1 recombinant vaccinia virus. These results demonstrate that HIV-B35-SF2-24 is a naturally processed peptide which is presented by both HLA-B*3501 and HLA-B*5101. TCR analysis of one CTL clone suggested that it is a single clone. B*3501-SF2-24-tetrameric complexes inhibited both HLA-B*3501- and HLA-B*5101-restricted recognition of this clone, suggesting that the TCR of this clone cross-recognize the structure of both HLA class I-peptide complexes. PMID- 11009086 TI - A global appraisal of immunodominant CD8 T cell responses to Epstein-Barr virus and cytomegalovirus by bulk screening. AB - Knowledge of the immunodominant responses to Epstein-Barr Virus (EBV) and human cytomegalovirus (HCMV) should help to generate cytotoxic T cell lines to these herpesviruses. Here we report on the analysis of CD8 T cell responses to EBV and HCMV in the blood of kidney transplant recipients undergoing viral reactivation (n = 16) and in healthy virus carriers (n = 10). We used a transient COS transfection assay that permits semi-quantitative estimation of CD8+ T cell responses against a larger number of HLA/viral protein combinations within polyclonal T cell lines and thus allows a rapid identification of major epitopes. From the comparison of these responses to those that we previously described in the synovial fluid of patients suffering from various forms of chronic arthritis (n = 32), it appears that EBV-specific T cells are mainly directed against a restricted set of immunodominant epitopes, primarily generated during the early lytic cycle and that both IE1 and pp65 are targets of the anti-HCMV response. We suggest that this method could be generally applied to the rapid identification of immunodominant T cell epitopes in viral and tumor immunity, and could help selecting HLA-peptide complexes that could be used to detect and sort specific T cell populations. PMID- 11009087 TI - The herpes simplex virus type 1 Fc receptor discriminates between IgG1 allotypes. AB - Herpes simplex virus type 1 (HSV-1) expresses a complex of two virally encoded glycoproteins, gE and gl, which is capable of binding nonimmune human IgG. The gE gl complex has thus become known as an Fc receptor (FcR), which reportedly binds human IgG subclasses in the order IgG4 > IgG1 > or = IgG2 and does not bind IgG3 from many individuals. There is, however, allelic variation in the genes encoding the human IgG1 heavy chain constant region and this gives rise to allotypes of IgG1. Using recombinant monoclonal IgG molecules of known isotype and mutants thereof we have unexpectedly discovered that the HSV-1 FcR discriminates between IgG1 allotypes. This is evidence of functional differences between IgG1 allotypes that may account for their distribution in populations. Furthermore, these findings suggest HSV-1 FcR binding sites on the IgG molecule some distance from the proposed binding site in the CH2-CH3 domain interface. PMID- 11009088 TI - Role of CD154 in the secondary immune response: the reduction of pre-existing splenic germinal centers and anti-factor VIII inhibitor titer. AB - Using the murine model of hemophilia A, we have examined the role of CD154 in the secondary immune response to factor VIII (FVIII). We previously reported that repeated i.v. injection of FVIII in hemophilia A mice induces a T cell-dependent anti-FVIII antibody formation. Herein, blocking of CD154 by a monoclonal antibody in FVIII-primed hemophilia A mice resulted in the disappearance of pre-existing spleen germinal centers (GC) in the white pulp within 24 h of treatment. Moreover, further expansion of GC in response to FVIII challenge was completely inhibited. In parallel, anti-FVIII antibody titers were markedly reduced and T cell responses to FVIII were abolished. The rapid disappearance of the GC after anti-CD154 treatment was not accompanied by increased B cell apoptosis; instead B cells accumulated in the peripheral zone of the splenic white pulp. Interestingly, repeated exposure to FVIII with anti-CD154 antibody administration blocked anti-FVIII antibody formation but failed to induce long-lasting unresponsiveness. Our data demonstrate that the CD40-CD154 interaction is critical for B cell homeostasis and the secondary immune response to FVIII. For potential clinical application, the data also suggest that therapies targeting the CD154 molecule may be useful for the treatment of high titer FVIII inhibitors in hemophilia A. PMID- 11009089 TI - Peroxovanadium-mediated protection against murine leishmaniasis: role of the modulation of nitric oxide. AB - The phosphotyrosine phosphatase inhibitor bpV(phen) has the ability to markedly decrease the progression of leishmaniasis in vivo. Here, we have identified the mechanisms that are responsible for this protective effect. We report that two potent peroxovanadium (pV) compounds, bpV(phen) and bpV(pic), control progression of leishmaniasis in a similar manner by modulating NO-dependent microbicidal action. We observed that their injection can rapidly and transiently induce the expression of inducible NO synthase (iNOS) in livers of mice and enhance circulating nitrate levels. Treatment of mice with bpV(phen) or bpV(pic) completely controlled progression of leishmaniasis in an NO-dependent manner, since inhibition of iNOS with aminoguanidine completely reversed this pV-mediated protection. This NO-dependent pV-mediated protection was further demonstrated by the incapacity of bpV(phen)-treated Nramp-/-, iNOS-/- mutant mice to control Leishmania major infection. Using an air pouch model, we showed that bpV(phen) can strongly modulate secretion of L. major-induced pro-inflammatory molecules and neutrophil recruitment. In addition, we observed that bpV(phen) per se can strongly induce the expression of Th1 type cytokines over Th2 in spleens of animals. Overall, this study has allowed us to establish the in vivo functional and immunological events involved in pV-mediated protective mechanism against leishmaniasis and that NO plays a pivotal role in this process. PMID- 11009090 TI - Interleukin-12 and Flt3 ligand differentially promote dendropoiesis in vivo. AB - Interleukin-12 (IL-12) and Flt3 ligand (FL) regulate hematopoiesis by stimulating proliferation, differentiation and migration of progenitor and/or stem cells. In this study, we evaluated the in vivo effect of IL-12 alone or in combination with FL on dendritic cell (DC) generation and accumulation in murine spleen, lymph node, thymus, liver, and tumor tissues. Both cytokines induced accumulation of functional DC in lymphoid and non-lymphoid tissues. IL-12 promoted predominantly myeloid dendropoiesis, while FL induced both myeloid and lymphoid dendropoiesis. Combination treatment resulted in a dramatic increase in CD86+, and particularly, NLDC-145+ cells within the liver, which was largely due to cell proliferation. Combination therapy also revealed the ability of FL to protect bone marrow cell populations from IL-12-induced depletion in vivo. In vitro, we found a significant FL-induced up-regulation of IL-12 production by DC at both mRNA and protein levels. Thus, our study suggests that (i) the antitumor activity of IL-12 may, at least in part, be mediated by the stimulation of dendropoiesis and (ii) IL-12 might contribute to the antitumor activity of FL. Furthermore, induction of DC generation in vivo by a combination of IL-12 and FL might become a new approach for immunotherapy of cancer. PMID- 11009091 TI - C/EBPbeta enhances IL-4 but impairs IL-2 and IFN-gamma induction in T cells. AB - C/EBP transcription factors have been described to control the activity of the human IL-4 promoter. The C/EBP binding sites within the IL-4 promoter overlap with composite NF-AT and AP-1 binding motifs. We show here that similar binding sites are part of the murine IL-4 promoter. Retroviral overexpression of C/EBPbeta in murine EL-4 thymoma cells led to a strong induction of endogenous IL 4 and a reduction in IL-2 and IFN-gamma expression. Similarily, in primary murine T cells C/EBPbeta induction resulted in an increase in IL-4 levels, whereas in human Jurkat T cells a decrease in IL-2 RNA was detected. Like AP-1, C/EBP factors belong to the large class of basic leucine zipper proteins. However, unlike AP-1, C/EBPbeta does not act in synergy with NF-AT in the induction of the murine IL-4 promoter. Instead, both factors compete in their binding to the P4/Pu bD site, one of the most important sequence elements of the IL-4 promoter. Whereas NF-AT factors require high levels of free Ca2+ and calcineurin activity for induction, C/EBP induction in T cells is Ca2+/calcineurin independent. These observations suggest that various induction conditions lead to the activation of transcription factors, inducing IL-4 promoter activity at specific developmental stages of T cells. PMID- 11009092 TI - Sex steroids induce apoptosis of CD8+CD4+ double-positive thymocytes via TNF alpha. AB - T cell production by the thymus, thymic size, cellularity and output all decrease drastically after puberty. Among the candidates that may mediate this decrease are the sex steroids: hypersecretion or pharmacological administration of these hormones has long been known to induce thymic hypocellularity, and their depletion yields thymic hypercellularity. Here we show that a typical sex steroid, testosterone, specifically targets CD8+CD4+ double-positive (DP) thymocytes for apoptosis via TNF-alpha. Anti-TNF-alpha monoclonal antibodies abrogated testosterone-induced DP apoptosis, and TNF-alpha-/- DP thymocytes were largely resistant to testosterone-mediated apoptosis in vivo. Testosterone accomplished this effect by upregulating TNF-alpha production and by simultaneously sensitizing DP thymocytes to TNF-alpha. Thus, TNF-alpha is the critical mediator of sex steroid-induced apoptosis in thymocytes, and its manipulation should provide a point of intervention to modulate T cell production in sex hormone disorders. PMID- 11009093 TI - Locally inducible CD66a (CEACAM1) as an amplifier of the human intestinal T cell response. AB - CD66a is an adhesion molecule member of the carcinoembryonic antigen immunoglobulin-like family present on the surface of epithelial cells, granulocytes and IL-2 activated T cells. We studied whether CD66a is expressed in vivo by T lymphocytes and whether it affects TCR-mediated activation. CD66a was detected by histochemistry, flow cytometry analysis, reverse transcription PCR and Western blot on fresh colon biopsies and T cell clones. A fraction of T cells in the lamina propria express CD66a, which is induced by IL-7 and IL-15 cytokines. T cells express four different CD66a splice variants and at least two forms of the protein are glycosylated in a cell type-specific manner. Triggering of CD66a on T cells with physiological ligands or with specific mAb increases TCR mediated lymphokine release, in an antigen dose-independent manner. This effect requires the presence of the CD66a intracytoplasmic domain, which contains two immunoglobulin receptor family tyrosine-based inhibitory motif-like domains, as shown by stimulation of Jurkat cells transfected with different CD66a isoforms and is associated with increased induction of AP1 and NFkappaB transcription factors. These data indicate that CD66a amplifies T cell activation and thus could facilitate crosstalk between epithelial cells and T lymphocytes in intestinal immune response. PMID- 11009094 TI - Identification of an IFN-gamma responsive region in an intron of the invariant chain gene. AB - The mechanism by which IFN-gamma up-regulates invariant chain mRNA in antigen presenting cells has been under intensive investigation. This study shows that in murine monocytic cells the transcriptional up-regulation mediated by the invariant chain (Ii) upstream enhancer only accounts for part of the induction of Ii mRNA by IFN-gamma. We have identified an intronic region in the murine Ii gene that contributes to the transcriptional up-regulation by IFN-gamma. The region has S (H), X/X2 and Y boxes similar to those in MHC class II promoters and the Ii upstream enhancer. Mutations at the putative S, X and Y boxes have abolished the ability of the region to mediate Ii up-regulation by IFN-gamma. Consistent with the functions of these boxes, our findings reveal that the up-regulation of Ii transcription by IFN-gamma mediated by the intronic region is dependent on the induction of class II transactivator by IFN-gamma. PMID- 11009095 TI - MHC class II and CD40 play opposing roles in dendritic cell survival. AB - In contrast to very immature dendritic cells (DC), mature DC are largely resistant to death by CD95 (CD95/APO-1) ligation. Investigation of other potential death-inducing ligands showed that mature DC were instead highly susceptible to apoptosis induced by cross-linking of MHC class II. Thus, increasing DC maturity correlates with increased resistance to CD95 killing, but an increased susceptibility to class II-mediated killing. Anti-I-A/I-E monoclonal antibodies (mAb) induced rapid (<2 h) apoptotic cell death in mature epidermal, spleen and bone marrow-derived DC, as determined by annexin/propidium iodide staining, morphological changes, decreased diploidy and loss in mitochondrial membrane potential. Although full class II-mediated killing required DC cytoskeletal motion, divalent cations and phosphatase activity, neither caspase activation, respiration, RNA or protein synthesis, NO production, nor CD95:CD95L interactions were required. Strikingly, DC pretreated by CD40 mAb cross-linking, but not by lipopolysaccharide or TNF-alpha, were completely resistant to class II mediated killing. CD40-mediated protection was reduced in the presence of the SB202190 inhibitor of the mitogen-activated protein kinase p38 pathway, but appeared to be independent of p42/44 extracellular signal-related kinase or NF-KB activation. Our findings show that in addition to its role as an activator of antigen-presenting cell function, CD40 provides an important counter-signal against class II-induced apoptosis. Thus, these data point to an important role of the T cell in regulating DC survival. PMID- 11009096 TI - Temporally regulated assembly of a dynamic signaling complex associated with the activated TCR. AB - TCR triggering promotes multiple tyrosine kinase-dependent interactions involving proteins with one or more protein binding modules. Reported interactions mostly exceed the binding potential of these proteins. A solution to this paradox is the temporally regulated recruitment of alternative ligands. We have tested this hypothesis by analyzing the time course of protein/protein interactions triggered by TCR engagement. We show that a short-lived and dynamic multimolecular complex is assembled on tyrosine-phosphorylated CD3zeta. Specific components of this complex are recruited and shed in a temporal sequence distinct for each of the proteins analyzed. The temporally regulated assembly of a higher order structure at the activated TCR is likely to be crucial in achieving both signal longevity and signal specificity. PMID- 11009097 TI - Defective recruitment and activation of ZAP-70 in common variable immunodeficiency patients with T cell defects. AB - We have previously identified a subset of common variable immunodeficiency (CVID) patients with defective T cell function associated with impaired activation of the TCR-dependent tyrosine phosphorylation cascade. Here we have assessed the structural and functional integrity of the principal components involved in coupling the TCR/CD3 complex to intracellular tyrosine kinases in two of these patients. We show that ZAP-70 fails to bind the signaling-competent CD3zeta tyrosine phosphorylation isoform and to become activated following TCR engagement, suggesting that defective recruitment of ZAP-70 might underlie the TCR signaling dysfunction in these patients. Determination of the nucleotide sequences encoding the intracellular domains of the CD3/zeta subunits and ZAP-70 did not reveal any mutation. Furthermore, ZAP-70 from these patients could interact in vitro with recombinant phospho-zeta, ruling out genetic defects at the immunoreceptor tyrosine-based activation motif/SH2 domain interface responsible for ZAP-70 recruitment to the activated TCR. No defect was found in expression, activity or subcellular localization of Lck, which is thought to be primarily responsible for CD3zeta phosphorylation. Hence, while the T cell defect in these CVID patients can be pinpointed to the interaction between ZAP-70 and CD3zeta, the integrity in the components of the signaling machinery involved in this process suggests that additional components might be required for completion of this step. PMID- 11009098 TI - TGF-beta1 down-regulates Th2 development and results in decreased IL-4-induced STAT6 activation and GATA-3 expression. AB - TGF-beta plays an important role in immune regulation in vivo and affects T cell differentiation in vitro. Here we describe how TGF-beta modulates Th2 development in vitro and investigate its mechanisms of action. TGF-beta down-regulated Th2 development of naive CD4+ Mel-14high T cells derived from the DO11.10 ovalbumin specific TCR-transgenic mouse, and this was observed both in cultures driven with anti-CD3 and anti-CD28 and with splenic APC and antigen. TGF-beta down-regulated GATA-3 expression in developing Th2 and these cells showed a diminished IL-4 induced STAT6 activation. We found, however, that naive cells driven in Th2 conditions with TGF-beta did not show a significantly decreased STAT6 activation, suggesting that TGF-beta inhibits Th2 development via a STAT6-independent mechanism. PMID- 11009099 TI - B7 co-stimulatory requirements differ for induction of immune responses by DNA, protein and recombinant pox virus vaccination. AB - Whether B7-1 and B7-2 have distinct functions for eliciting immune responses to antigens that are presented to the immune system by intracellular and extracellular antigen processing pathways is an unresolved question. To investigate this issue we compared the humoral and cellular immune responses elicited by immunizing wild-type, B7-1-/- and B7-2-/- mice with either HIV-1 gp120 plasmid DNA, recombinant gp120 protein or vaccinia virus expressing gp120. The generation of both humoral and cellular immune responses to an antigen produced intracellularly following DNA vaccination had critical requirements for B7-2, but not B7-1. Neither of the molecules was essential for the generation of antibody responses to an extracellular protein antigen administered with adjuvant; B7-1 had little effect on the elicited immune responses. When recombinant vaccinia virus was used to present antigen intracellularly in the context of a viral infection, B7-2 was absolutely required for antibody and T cell proliferative responses, but it exerted a suppressive effect on the elicited CTL activity. These results demonstrate that antigens presented to the immune system by different mechanisms have distinct B7-1 and B7-2 co-stimulatory requirements. PMID- 11009100 TI - MMCP-8, the first lineage-specific differentiation marker for mouse basophils. Elevated numbers of potent IL-4-producing and MMCP-8-positive cells in spleens of malaria-infected mice. AB - In mice infected with the non-lethal malaria parasite Plasmodium chabaudi chabaudi AS, a prominent switch from a Th1 to a Th2 type of response occurs in CD4+ T cells at the time of peak parasitemia or shortly thereafter (9-15 days after infection). This is accompanied by a major increase in IL-4, and a similar decrease in IFN-gamma-producing cells. Non-B-non-T cells have been shown to be the main source of the IL-4 in these mice. The IL-4-producing cells are hyperresponsive to IL-3, indicating mast cell or basophil origin. To further characterize this cell population we have studied various organs at different time points of malarial infection by Northern blot analysis. No significant increase in the expression of any of the classical mouse mast cell serine proteases (MMCP)-1 to 7 or carboxypeptidase A was detected in the spleen during the entire infection. However, a marked increase in the expression of MMCP-8 was observed in the spleen at around day 15 post infection. Isolation of IgE receptor positive cells from spleen shortly after peak parasitemia led to a prominent enrichment of MMCP-8-expressing cells. Fifty thousand of these cells were, after IL-3 stimulation, found to produce IL-4 to levels comparable with more than one million fully activated T cells. Our results show that basophil-like cells are very potent producers of IL-4 and that IL-4 produced by these cells may be of major importance for the initiation of a Th2 response. In addition, the detection of MMCP-8 in these cells has led to the identification of the first basophil specific differentiation marker in the mouse. PMID- 11009101 TI - Alternatively activated macrophages induced by nematode infection inhibit proliferation via cell-to-cell contact. AB - The cytokine microenvironment is thought to play an important role in the generation of immunoregulatory cells. Nematode infections are commonly associated with Th2 cytokines and hyporesponsive T cells. Here we show that IL-4-dependent macrophages recruited in vivo by the nematode parasite Brugia malayi actively suppress the proliferation of lymphocytes on co-culture in vitro. These alternatively activated macrophages block proliferation by cell-to-cell contact, implicating a receptor-mediated mechanism. Further, the proliferative block is reversible and is not a result of apoptosis. Suppressed cells accumulate in the G1 and G2/M phase of the cell cycle. Interestingly, the G1 and G2/M block correlates with increased levels of Ki-67 protein, suggesting a mechanism that affects degradation of cell cycle proteins. We also show that, in addition to lymphocyte cell lines of murine origin, these suppressive cells can inhibit proliferation of a wide range of transformed human carcinoma lines. Our data reveal a novel mechanism of proliferative suppression induced by a parasitic nematode that acts via IL-4-dependent macrophages. These macrophages may function as important immune regulatory cells in both infectious and noninfectious disease contexts. PMID- 11009102 TI - The synthetic, oxidized C-terminal fragment of the Plasmodium berghei circumsporozoite protein elicits a high protective response. AB - A polypeptide of 69 amino acids (PbCS 242-310) encompassing the C-terminal region of the circumsporozoite protein of Plasmodium berghei (PbCS) was generated using solid-phase peptide synthesis. The immunological and protective properties of peptide PbCS 242-310 were studied in BALB/c mice (H-2d). Two subcutaneous injections, in the presence of IFA at the base of the tail, generated (i) high titers of anti-peptide antibodies which also recognized the native P. berghei CS protein, (ii) cytolytic T cells specific for the Kd-restricted peptide PbCS 245 253 and (iii) partial CD8+-dependent protection against sporozoite-induced malaria. The same frequencies of peptide PbCS 245-253 specific CD8+ T cells were found by IFN-gamma ELISPOT in the draining lymph nodes of animals immunized with the short optimal CTL peptide 245-253 or with the polypeptide 242-310, indicating that the longer polypeptide can be processed and presented in vivo in the context of MHC class I as efficiently as the short CTL peptide. Interestingly, higher levels of IFN-gamma producing CD8 T cells and protection were observed when the four cysteine residues present in the C-terminal peptide were fully oxidized. These findings underline the potential importance of the chemical nature of the C terminal fragment on the activation of the immune system and concomitant protection. PMID- 11009103 TI - Regulation of cell survival during B lymphopoiesis: increased pre-B cell apoptosis in CD24-transgenic mouse bone marrow. AB - CD24 (heat-stable antigen) is expressed in a developmentally regulated fashion by B cell precursors in mouse bone marrow (BM), but its role in B lymphopoiesis remains obscure. A slight overexpression of CD24 in transgenic (Tg) mice leads to depletion of B lymphoid cells in BM. The present study examines whether CD24 is involved in apoptotic selection of B lineage cells under normal microenvironmental conditions in vivo. Double immunofluorescence labeling and flow cytometry have been used to quantitate the apoptotic rates of phenotypically defined B cell populations in BM of CD24-Tg mice. Apoptosis of pre-B cells expressing cytoplasmic mu heavy chains of IgM but lacking surface (s)IgM was increased both ex vivo and in short-term culture, while the number of pre-B cells was halved compared to BM of normal mice. In contrast, B220+mu- pro-B cells and sIgM+ B lymphocytes showed no significant change in either apoptosis or number. The findings provide evidence that CD24 can play a role in vivo in modulating pre B cell apoptosis, a quality control checkpoint in B cell development. PMID- 11009104 TI - A disulfide-linked natural killer cell receptor dimer has higher affinity for HLA C than wild-type monomer. AB - Inhibitory receptors on the surface of natural killer (NK) cells recognize specific MHC class I molecules on target cells and prevent the target cell lysis by NK cells. The killer cell immunoglobulin-related receptors (KIR), KIR2D, found in human, specifically interact with polymorphic HLA-C molecules. The crystal structure of the inhibitory receptor, KIR2DL1, revealed a relationship to the hematopoietic receptor family, suggesting that the signaling mechanism of KIR2D molecules may resemble that of the hematopoietic receptors, and involve KIR2D dimerization. We have engineered a disulfide-linked dimer of KIR2DL1 by introducing a free cysteine at the C-terminal stem region of the receptor. The disulfide-linked KIR2DL1 dimer binds to HLA-Cw4 at a molar ratio of one dimer to one HLA-Cw4 molecule. Furthermore, the covalently-linked KIR2DL1 dimer binds more tightly to HLA-Cw4 than the wild-type monomer, suggesting the occurrence of a second binding event that increases the overall affinity of KIR dimer for HLA-C. PMID- 11009105 TI - Interleukin-4 transgenic mice develop glomerulosclerosis independent of immunoglobulin deposition. AB - Mice with constitutive transgenic (tg) expression of IL-4 develop autoimmune-type disorders resembling human lupus nephritis. The kidneys show progressive glomerulosclerosis with immunoglobulin (Ig) and complement deposition. This study investigated the roles of renal IL-4 expression and glomerular Ig deposition in the pathogenesis of glomerulosclerosis in IL-4 tg mice. Treatment of these mice with IL-4 neutralizing antibody prevented renal disease. IL-4 tg mice treated with methylprednisolone (MP) showed increased mesangial collagen deposition with only trace amounts of glomerular Ig. To analyze the relevance of Ig deposition in the development of the renal lesions, IL-4 tg mice were cross-bred with mu chain deficient mice (muMT-/-), which are unable to produce Ig. IL-4 tg/muMT-/- mice developed progressive glomerulosclerosis with mesangial accumulation of collagen types I, IV and V despite complete absence of glomerular Ig deposits. Renal IL-4 expression was observed in both anti-IL-4- and MP-treated IL-4 tg mice as well as in IL-4 tg/muMT-/- mice. No statistical difference in the number of glomerular T cells and macrophages between any of the groups was evident. Our data demonstrate that in this model glomerulosclerosis can develop independently of and prior to Ig deposition, and suggest that the initial accumulation of glomerular extracellular matrix is due to renal IL-4 expression. Our results point to a novel mechanism for the development of glomerulosclerosis which may have implications for human disease. PMID- 11009106 TI - CD8 memory, immunodominance, and antigenic escape. AB - Previous theoretical work has suggested that efficient virus control or clearance requires antigen-independent persistence of memory cytotoxic T lymphocyte precursors (CTLp), and that failure to generate such memory CTLp can result in persistent infection and eventual loss of virus control. Here we use mathematical models to investigate the relationship between virus control, the clonal composition of the CTL response and the chance of the virus to evolve antigenic escape. In the presence of efficient memory CTLp, virus is controlled at very low levels by a broad CTL response directed against multiple epitopes. In this case, antigenic escape of the virus population is expected to take a very long time. On the other hand, if the CTL response is short lived in the absence of antigen, virus replicates at higher levels and is only opposed by a narrow CTL response, characterized by an immunodominant CTL clone. In this case, antigenic escape is expected to evolve in a short period of time, resulting in progressive loss of virus control. We discuss our findings in relation to data from HIV-1-infected patients. PMID- 11009107 TI - High frequencies of circulating IFN-gamma-secreting CD8 cytotoxic T cells specific for a novel MHC class I-restricted Mycobacterium tuberculosis epitope in M. tuberculosis-infected subjects without disease. AB - MHC class I-restricted CD8 cytotoxic T lymphocytes (CTL) are essential for protective immunity to Mycobacterium tuberculosis in animal models but their role in humans remains unclear. We therefore studied subjects who had successfully contained M. tuberculosis infection in vivo, i.e. exposed healthy household contacts and individuals with inactive self-healed pulmonary tuberculosis. Using the ELISPOT assay for IFN-gamma, we screened peptides from ESAT-6, a secreted antigen that is highly specific for M. tuberculosis. We identified a novel nonamer epitope: unstimulated peripheral blood-derived CD8 T cells displayed peptide-specific IFN-gamma release ex vivo while CD8 T cell lines and clones exhibited HLA-A68.02-restricted cytolytic activity and recognized endogenously processed antigen. The frequency of CD8 CTL specific for this single M. tuberculosis epitope, 1/2500 peripheral blood lymphocytes, was equivalent to the combined frequency of all IFN-gamma-secreting purified protein derivative reactive T cells ex vivo. This highly focused CTL response was maintained in an asymptomatic contact over 2 years and is the most potent antigen-specific antimycobacterial CD8 CTL response hitherto described. Thus, human M. tuberculosis-specific CD8 CTL are not necessarily associated with active disease per se. Rather, our results are consistent with a protective role for these ESAT 6-specific CD8 T cells in the long-term control of M. tuberculosis in vivo in humans. PMID- 11009108 TI - CD45R, CD44 and MHC class II are signaling molecules for the cytoskeleton dependent induction of dendrites and motility in activated B cells. AB - Anti-CD44 or anti-MHC II antibodies bound to tissue culture plates have previously been shown to induce a dramatic generation of dendritic processes in activated murine B cells. In this study, we demonstrate a similar generation of dendrites and cell motility in activated B cells through CD45R. The dynamic formation of dendritic processes and associated induction of cell motility were analyzed by video microscopy and were characterized by a rapid, and multidirectional emission of dendrites with retractile behavior. The addition of cytochalasin E totally blocked dendrites formation and motility induced through either CD45R, CD44 or MHC II, suggesting that the necessary cytoskeletal rearrangements require active polymerization of actin. Confocal microscopy showed an accumulation of F-actin in the dendrites, as long as cells were elongating. In contrast, G-actin was localized in the perinuclear area and also accumulated in sites where dendrites originated. Preincubation of B cells with staurosporine (a PKC inhibitor) or BAPTA-AM (a calcium chelator) prevented these morphological changes, indicating additionally a requirement for a PKC-calcium-dependent activity. Dendrite formation and cellular motility, therefore, seem to be two manifestations of the same phenomenon, and CD44, CD45R and MHC II appear to be signaling molecules for the observed cytoskeleton-dependent morphological changes. PMID- 11009109 TI - In experimental leishmaniasis deficiency of CD18 results in parasite dissemination associated with altered macrophage functions and incomplete Th1 cell response. AB - We investigated experimental leishmaniasis in CD18-deficient mice. Whereas wild type (WT) CD18-/- mice (129SV/C57BL/6) were resistant to infection, CD18-/- mice revealed increasing visceral dissemination of parasites. Unlike in other susceptible strains, infected footpads of CD18-/- mice did not ulcerate, due to an abolished recruitment of granulocytes. In vitro, CD18-/- macrophages were able to phagocytose opsonized Leishmania major despite absence of CR3, albeit phagocytosis rate was 50% lower than in WT macrophages. We found that uptake was partially mediated by scavenger receptors. As infected CD18-/- macrophages showed impaired ability to produce NO and to eliminate parasites, CD18 is one mediator of NO production. CD18 is also involved in reduction of IL-12 release by L. major infected macrophages, as uptake of opsonized parasites (via CR3) decreased IL-12 release only in WT, but not in CD18-/- macrophages. When T cells from infected CD18-/- mice were restimulated with antigen-presenting cells (APC), they released no IL-2 or IL-4, but a little IFN-gamma, associated with lack of proliferation. This deficiency was linked to absence of CD18 on T cells, but not on APC. Substitution with IL-2 specifically restored a Th1-like response with proliferation and release of IFN-gamma. Thus, while impaired phagocytosis, NO production, and recruitment of granulocytes in CD18-/- mice may not reverse resistance, and while unrestricted IL-12 release supports development of Th1 cells, the failure of T cells to release IL-2 and to proliferate causes susceptibility. PMID- 11009110 TI - Removal of parvovirus B19 from hemoglobin solution by nanofiltration. AB - Parvovirus B19 (B19), which may contaminate red cell components for blood transfusion, is known to be resistant to several viral inactivation methods. To increase the safety of hemoglobin solutions as a source of red cell substitutes, we investigated the removal of parvovirus B19 from hemoglobin solution using nanofiltration. The hemoglobin solution spiked with parvovirus B19 was tangentially filtered using the BMM-35 filter (mean pore size of 35 nm) followed by BMM-15. The parvovirus B19 of 10(8.5) PT50 (median PCR titer)/10 microL was not changed after the BMM-35 filtration. However, the BMM-15 filtration decreased the parvovirus B19 from 10(8.3-8.7) PT50 to 10(1.3-2.2) PT50, indicating more than 6 log10 reduction. When the initial parvovirus B19 of 10(6.0) PT50 was subjected to the BMM-15 filtration, the residual virus was 10(-0.3-0.5) PT50 orundetected in some fraction of the filtrate. Hemoglobin recovery was 70.4 +/- 3.4%. The ratio of methemoglobin was not changed during the filtration. These findings indicate that the BMM-15 filtration is a promising approach to prepare a safer hemoglobin solution for red cell substitutes. PMID- 11009111 TI - Effect of alpha-alpha diaspirin crosslinked hemoglobin (DCLHb) on the potency of sodium nitroprusside and nitroglycerine to decrease blood pressure in rats: a dose-response study. AB - The nitrovasodilators, sodium nitroprusside and nitroglycerine, effect a dose dependent decrease in mean arterial blood pressure (MABP) by liberating nitric oxide. Alpha-alpha diaspirin crosslinked hemoglobin (DCLHb) is known to bind nitric oxide. We studied the effect of DCLHb on the potency of sodium nitroprusside (n=36) and nitroglycerine (n=36) to decrease MABP in rats which received 1, 10, 100, 1,000, or 10,000 mg/kg of the DCLHb, or normal saline as the Control. Six doses of sodium nitroprusside or nitroglycerine were given to each rat in a systematically varied sequence. For both drugs, in rats given 1, 10, or 100 mg/kg of DCLHb there were no between groups differences in the change in MABP compared to the Control group. For rats that received 1,000 or 10,000 mg/kg of DCLHb, the potency of nitroglycerine and sodium nitroprusside to decrease MABP was less (p<0.05) than the other groups. These data support the hypothesis that small doses of DCLHb effect a minimal change in the potency of nitrovasodilators to reduce blood pressure. However, they suggest that clinically relevant doses of DCLHb attenuate the vasodilatory ability of sodium nitroprusside and nitroglycerine. PMID- 11009113 TI - A novel immunoadsorbent for rheumatoid arthritis therapy--preparation and efficacy evaluation. AB - AIM: To develop a novel immunoadsorbent for rheumatoid arthritis (RA) therapy. METHODS: A RA immunoadsorbent was developed by binding heat-aggregated human IgG(HAHIgG) to porous agar gel beads. Its adsorption capacity for rheumatoid factors (RFs), storage stability and blood compatibility were evaluated. RESULTS: The coupling yield of HAHIgG on the carrier was 6.0 mg/g wet gel. Saturation adsorption capacity of the adsorbent for IgMRF, IgGRF and IgARF were 3400, 2240 and 2400 IU/g, respectively. The adsorbent can be stored at 4 degrees C for three months without significant variance in its activity. Its fine permeability and hemocompatibility were demonstrated by extracorporeal hemoperfusion on rabbits. CONCLUSION: HAHIgG/agar gel is a safe and effective immunoadsorbent for RA therapy, its potential clinical use is promising in the future. PMID- 11009112 TI - Artificial cells microencapsulated genetically engineered E. coli DH5 cells for the lowering of plasma creatinine in-vitro and in-vivo. AB - High level of plasma creatinine occurs in renal insufficiency, uremia, and other diseases. At present lowering of this metabolite is done by using dialysis and other techniques. In this article, we report the use of artificial cells microencapsulated genetically engineered E. coli DH5 cells for lowering plasma creatinine in-vitro and in-vivo. Result shows that artificial cells were able to lower plasma creatinine in-vitro from 21.80 +/- 1.10 mg/dl to 21.80 +/- 1.10 mg/dl in 60 minutes and to 19.34 +/- 0.60 mg/dl in 3 hours. Result also shows that when given orally on a daily basis, artificial cells microencapsulated genetically engineered E. coli DH5 cells were also able to lower plasma creatinine in rats. PMID- 11009114 TI - Effects of liposome-encapsulated ciprofloxacin on phagocytosis, nitric oxide and intracellular killing of Staphylcoccus aureus by murine macrophages. AB - The effects of liposome-encapsulated ciprofloxacin on phagocytosis, nitric oxide production and intracellular killing of Staphylococcus aureus in murine macrophages were evaluated in this study. Mice were pretreated with three daily doses of liposome-encapsulated ciprofloxacin (45 mg/kg body weight/dose, intraperitoneal injection). At day 3 post drug administration, peritoneal macrophages were harvested by peritoneal lavage, and the phagocytic activity of the macrophages was determined by a chemiluminescence assay using opsonized zymosan particles. The phagocytic activity was found to be 7-fold higher in the liposome-encapsulated ciprofloxacin-treated group when compared to the untreated control group. For S. aureus-infected macrophages incubated with liposomes containing subinhibitory concentrations of ciprofloxacin (0.05 to 0.25 microg/mL), there were significant increases (up to 40 microM) in the levels of nitrite (NO2-, an end product of nitric oxide synthesis), and concommitant decreases (2-3 log) in the intracellular concentrations of S. aureus. Peak nitrite levels (20-40 microM) were produced when concentrations of liposome encapsulated ciprofloxacin used were 0.1 to 0.25 microg/mL. These results suggest that liposome-encapsulated ciprofloxacin may have profound effects on the immunological functions of macrophages. PMID- 11009115 TI - Plasmapheresis in life-threatening verapamil intoxication. AB - Verapamil intoxications are life-threatening conditions with a far too often fatal outcome. In 2 patients, severe suicidal intoxication by 2.4 g and 9.6 g of verapamil orally resulted in life threatening hypotension and bradycardia with the need of heart-pacing and resuscitation. Plasmapheresis was started within less then 4 hours after intoxication and seemed to reduce the verapamil plasma concentration to less then 40%. A dramatic improvement of cardiovascular stability was already observed during plasmapheresis. In-vitro plasmapheresis was performed to verify the effectiveness of the extracorporeal detoxification. Verapamil was removed out of the blood by a clearance of 29.2 ml/min at blood flow of 200 ml/min. In conclusion, severe verapamil poisoning should be treated by early aggressive gut decontamination and an appropriate management of the haemodynamic complications. In case of lack of effectiveness for stabilisation, plasmapheresis can reduce verapamil related life threatening symptoms and bridge the time for the hepatic detoxification. PMID- 11009117 TI - Preventing scientific misconduct--times have changed. PMID- 11009116 TI - Assessment of low-power laser biostimulation on chondral lesions: an "in vivo" experimental study. AB - The purpose of this study was to evaluate whether intraoperative laser biostimulation can enhance healing of cartilaginous lesions of the knee. Surgery was performed on eighteen rabbits: a bilateral chondral lesion of 1.25 +/- 0.2 mm in length and 0.8 +/- 0.2 mm in width was created in the femoral medial condyle with a scalpel. The lesion in the left knee of each animal was treated intraoperatively using the diode Ga-Al-As 780nm. laser (300 Joules/cm2, 1 Watt, 300 Hertz, 10 minutes), while the right knee was left untreated, as control group. The animals were divided into three groups, A, B and C, according to the survival time after surgery, two, six and twelve weeks, respectively. The explants from the femoral condyles, both treated employing laser energy and left untreated, were examined histologically. Results showed a progressive filling with fibrous tissue of the cartilaginous lesion treated with laser irradiation, while no changes in the original lesion of the untreated group were observed at the end of the study. Maybe, in this experimental research, underexposure to laser irradiation was the cause for the absence of the necessary conditions for biostimulation. PMID- 11009118 TI - Alternative technique for medication injections. PMID- 11009119 TI - Effects of surgery type and attention focus on children's coping. AB - BACKGROUND: How children cope with minor or major surgery is influenced by their attention focus appraisals. Although factors that predict children's coping with surgery have been identified (i.e., age, locus of control, parent-doctor information, worry), it still is not known whether the type of surgery per se affects the coping strategies used and influences previously established predictors of coping. Furthermore, questions remain concerning the relation among type of surgery, attention focus, and coping. OBJECTIVES: The purpose of this study was to determine whether the type of surgery (minor vs. major) would have a differential effect on coping, and whether coping can be predicted better if it is known what type of attention focus (appraisal) the child has. METHODS: Data from three studies of children (n = 189) undergoing minor or major surgery were combined to examine the effects that type of surgery and attention have on coping. Measures included the Preoperative Mode of Coping Interview, Locus of Control Scale for Children, Parent-Doctor Information Interview, and a measure for worry. RESULTS: The results showed that the factors previously found to predict coping were upheld in the combined sample and accounted for 50% of the variance in coping. Type of surgery was significantly associated with coping: Children undergoing minor surgery were somewhat more vigilant than children undergoing major surgery. The inclusion of attention in the analysis significantly improved the variance explained in coping (66%), and children who had a concrete-objective focus of attention were found to be more vigilant. Significant interactions were found between attention focus and type of surgery, locus of control, and age. Type of surgery also had a significant interaction with worry. Children who focused on the concrete-objective aspects of the situation were more vigilant if they were having minor rather than major surgery. Also, children who had an internal locus of control and a concrete-objective focus of attention were more vigilant in coping. Regardless of age, children who had a concrete-objective focus of attention were more vigilant. Furthermore, at low levels of worry, children undergoing major surgery were more vigilant than children undergoing minor surgery. CONCLUSIONS: Coping with surgery is influenced by multiple factors. Children's ability to focus attention on the concrete objective aspects of surgery may help to reduce feelings of threat that could impede their use of vigilant coping. PMID- 11009120 TI - Hospital and emergency department use by young low-income children. AB - BACKGROUND: Limited data are available concerning determinants of health care service usage by low-income young children. OBJECTIVES: To explore predictors of hospitalization and emergency department (ED) use by young children of low-income families by using the Aday and Andersen Access Framework. METHODS: Low-income women (n = 474) with a child younger than 6 years completed a structured face-to face interview at human service offices or Women, Infants, and Children (WIC) clinics in four central Ohio counties. Women were considered low-income if they or their child were Medicaid eligible or uninsured. Data were collected for both the mother and the index child on sociodemographic status, health services use, health status, and access to care. RESULTS: Fifteen percent of the children had been hospitalized the previous year, and half had an ED visit. Hospitalization was significantly related to maternal hospitalization the previous year (OR = 2.5), child age younger than 1 year old (OR = 2.1) and more than two chronic conditions (OR = 2.2). Maternal ED usage in the last year (OR = 2.2), Medicaid fee for service plan (OR = 1.7), and rural residence (OR = 2.0) were predictive of ED use. CONCLUSIONS: Predisposing characteristics (maternal hospital/ED use) were predictive of both hospitalization and ED use by the index child. Enabling characteristics (fee-for-service Medicaid plan, rurality) were only predictive of ED use, and need characteristics (child's health) were only predictive of hospitalization. Further research to explore linkages between maternal and child use of health care services as well as the effect of changes in health care access, managed care, and other innovations on hospitalization and ED use in young, low-income children is recommended. PMID- 11009121 TI - Self-esteem and optimism in men and women infected with HIV. AB - BACKGROUND: Self-esteem and optimism have been associated with appraisal and outcomes in a variety of situations. The degree to which the contribution of self esteem and optimism to outcomes over time is accounted for by the differences in threat (primary) or resource (secondary) appraisal has not been established in persons with human immunodeficiency virus (HIV). OBJECTIVES: To examine the longitudinal relationship of personality (self-esteem and optimism) on primary and secondary appraisal and outcomes of well-being, mood, CD4+ T-lymphocyte count, and selected activities. METHODS: Men (n = 56) and women (n = 42) infected with HIV completed eight self-report measures twice over 18 months. Hierarchical Multiple Regressions were used to examine the relationship of personality variables on appraisals and outcomes. The mediating effects of primary and secondary appraisals were explored. RESULTS: Self-esteem uniquely accounted for 6% of the variance in primary appraisal and 5% in secondary appraisal. Optimism accounted for 8% of the unique variance in secondary appraisal. Primary and secondary appraisal mediated differently between personality and outcome variables. A strong predictor of well-being, mood disturbance, and activity disruption at Time 2 was participants' initial level of these variables. Socioeconomic status was a strong predictor of mood. CONCLUSIONS: Self-esteem and optimism are important but different resources for adapting to HIV disease. Strategies for reducing threats and increasing resources associated with HIV may improve an individual's mood and sense of well-being. PMID- 11009122 TI - Postpartum Depression Screening Scale: development and psychometric testing. AB - BACKGROUND: Approximately 400,000 mothers in the United States experience postpartum depression each year. However, only a small proportion of these women are identified as depressed by health care professionals. OBJECTIVES: To improve detection of this postpartum mood disorder, the purpose of this study was to assess the psychometric properties of a newly devised instrument, the Postpartum Depression Screening Scale (PDSS), a 35-item Likert-type self-report instrument. METHODS: Content validity was supported through the literature and the judgments rendered by a panel of five content experts and a focus group. The PDSS was administered to 525 new mothers. RESULTS: Confirmatory factor analysis provided empirical support for the existence of the hypothesized seven dimensions. A Tucker-Lewis goodness-of-fit index of 0.87 and a root mean square residual of 0.05 were judged supportive of model fit. Item response theory techniques provided further construct validity support for finer interpretations of the respective seven dimensions. Analysis of the Likert 5-point response categories further supported meaningful score interpretations. Alpha internal consistency reliabilities ranged from 0.83 (sleeping/eating disturbances) to 0.94 (loss of self). CONCLUSIONS: Empirically, all of the reliability and validity analyses supported the score interpretations posited for the PDSS. Currently, the sensitivity, specificity, and positive predictive value of the PDSS are being determined. PMID- 11009123 TI - The effect of structured training on breast self-examination search behaviors as measured using biomedical instrumentation. AB - BACKGROUND: Although breast self-examination (BSE) has long been recommended by health care practitioners as a complement to mammography and clinical breast examination, only a small percentage of U.S. women report doing monthly BSE, and an even smaller number of women perform this procedure proficiently. OBJECTIVES: To measure the effect of a structured training protocol on improving two dimensions of BSE technique (depth of palpation and search time) in each of two search patterns (vertical strip and concentric circle) using biomedical instrumentation. METHODS: For this study, 41 young women participated in a structured training protocol for BSE instruction. The dependent variable was thoroughness of search, for which there were two measures: depth of palpation (displacement of the sensors) and duration of the examination. An instrumented breast model designed by the investigator provided quantitative measurements of examination behaviors and was used to test outcomes of the instruction. RESULTS: Multivariate analyses demonstrated an overall difference across examinations (F = 28.03; p = 0.0001). Univariate tests showed treatment effects for both dependent variables: depth of palpation and duration of examination. CONCLUSIONS: Individual training in BSE with guided practice improved two measures of thoroughness of search: depth of palpation and duration of search time. Biomedical instrumentation represented a novel approach to the collection of quantitative performance data. PMID- 11009124 TI - Nonlinear components of age-related change in sleep initiation. AB - BACKGROUND: Although primary studies suggest that ability to initiate sleep declines as people age, no systematic literature review has addressed the age(s) at which adults experience the greatest change in their ability to initiate sleep. OBJECTIVE: To explore whether there are any points in time across the adult life span when the rate of change in ability to initiate sleep increases or decreases. METHODS: Mathematical modeling was used to generate data points from information about central tendency, variance, and correlations between age and time to sleep onset provided by seven research reports. The reports represent 258 subjects ages 17 to 91 years. Smoothing splines were used to identify inflection points suggestive of major changes in sleep initiation across the life span. RESULTS: Two mathematical models were generated. One model suggested that inflection points may exist around ages 30 and 50 years, respectively. With this model, the amount of time until sleep onset increased until the age of 30 years, but was unchanged from ages 30 to 50 years. Ability to initiate sleep appeared to decline steadily after the age of 50 years. The second model, with a p value of 0.05, lacked adequate power to identify a significant nonlinear trend. CONCLUSIONS: Decline in ability to initiate sleep may not occur at a steady rate over the adult life span. Further research is needed to pinpoint thresholds of change and possible gender differences in thresholds. PMID- 11009125 TI - Methodologic issues associated with secretion weight as a dependent variable in research using closed-system suction catheters. AB - BACKGROUND: Closed-system suction catheters (CSSC) were designed to eliminate the need to disconnect the patient from the ventilator during endotracheal suctioning (ETS). During data collection on an NIH-funded study, it was noted that moisture accumulated on the inside of the CSSC and sleeve when attached to the patient for 30 minutes. Because CSSC are not disconnected, they present unique methodologic problems related to measurement of secretions as a dependent variable in clinical research. OBJECTIVES: To describe a valid, reliable, and practical method for weighing secretions obtained during ETS using a CSSC; and to determine the change in weight of a CSSC after its attachment for 30 minutes to a mechanically ventilated patient. METHODS: After being weighed, a CSSC and sputum trap were attached to the endotracheal tube of a mechanically ventilated adult and remained attached for 43 minutes (30 minutes to allow positive end expiratory pressure and oxygenation levels to return to normal and 13 minutes to mimic the time frame for the ETS procedure used in another study). No ETS occurred. The CSSC and sputum trap were then removed and reweighed. RESULTS: A convenience sample consisted of 50 adults who were critically ill and mechanically ventilated. Independent variables included tidal volume, pressure support, body temperature, and respiratory rate. The dependent variable was wet weight of the CSSC, determined by subtracting the preprotocol catheter weight from the postprotocol catheter weight. The mean wet weight for all catheters was 0.5142 +/- 0.1215 grams. In a subset of 37 patients, two wet weights (74 paired observations) were determined. The mean wet weight for these catheters was 0.54014 +/- 0.1404 grams. The paired wet weights were statistically different (t = 2.433; df = 36; p = 0.02). Pearson correlation coefficients and beta coefficients were computed. While tidal volume and pressure support were highly correlated (r = 0.678; p = 0.011), there were no other statistically significant associations. CONCLUSIONS: The amount of secretions is a common dependent variable in ETS research. During the time that CSSC are attached to the patient and ventilator, moisture from either the ventilator's humidification system or the patient accumulates in the CSSC. This wet weight is not actually part of the secretions retrieved during ETS and should not be considered in the actual weight of secretions. Further study on the determinants of wet weight is warranted. PMID- 11009126 TI - Application of the theory of gender and power to examine HIV-related exposures, risk factors, and effective interventions for women. AB - Developed by Robert Connell, the theory of gender and power is a social structural theory based on existing philosophical writings of sexual inequality and gender and power imbalance. According to the theory of gender and power, there are three major social structures that characterize the gendered relationships between men and women: the sexual division of labor, the sexual division of power, and the structure of cathexis. The aim of this article is to apply an extended version of the theory of gender and power to examine the exposures, social/behavioral risk factors, and biological properties that increase women's vulnerability for acquiring HIV. Subsequently, the authors review several public health level HIV interventions aimed at reducing women's HIV risk. Employing the theory of gender and power among women marshals new kinds of data, asks new and broader questions with regard to women and their risk of HIV, and, most important, creates new options for prevention. PMID- 11009127 TI - New kinds of data, new options for HIV prevention among women: a public health challenge. PMID- 11009128 TI - Emergency department staff responses to a protocol of care for abused women. AB - While there is increasing recognition by health care providers of the need to identify and intervene in cases of partner abuse, there has been little evaluation of the interventions that have been implemented. This article documents emergency department staff responses to a five-step protocol of care on partner abuse implemented in a public hospital. It draws on in-depth interviews with a range of emergency department staff members, undertaken as part of the project's process evaluation. Findings suggest protocol training provided staff members with an opportunity to gain a greater awareness and understanding of partner abuse. Protocol use improved and consolidated existing practices; however, staff members were concerned about screening all women for abuse and highlighted areas in which further support and resources are required. Taking these factors into account may assist the appropriate and effective introduction of such protocols in other health care settings. PMID- 11009129 TI - A meta-analysis of fear appeals: implications for effective public health campaigns. AB - The fear appeal literature is examined in a comprehensive synthesis using meta analytical techniques. The meta-analysis suggests that strong fear appeals produce high levels of perceived severity and susceptibility, and are more persuasive than low or weak fear appeals. The results also indicate that fear appeals motivate adaptive danger control actions such as message acceptance and maladaptive fear control actions such as defensive avoidance or reactance. It appears that strong fear appeals and high-efficacy messages produce the greatest behavior change, whereas strong fear appeals with low-efficacy messages produce the greatest levels of defensive responses. Future directions and practical implications are provided. PMID- 11009130 TI - GO GIRLS!: results from a nutrition and physical activity program for low-income, overweight African American adolescent females. AB - This article describes the development, implementation, and results of an intervention designed for inner-city, overweight African American adolescent women. Fifty-seven participants were recruited from four public housing developments. Participants were administered physiologic, dietary, and cognitive assessments at baseline and immediately postintervention. Each session comprised three elements: (1) an educational/behavioral activity, (2) 30 to 60 minutes of physical activity, and (3) preparation and tasting of low-fat meals. In the absence of a control group, results were compared for high and low attenders, the former defined as attending at least 50% of the sessions. High attenders (n = 26) showed more favorable 6-month posttest values for most outcomes compared with low attenders (n = 31). These effects achieved statistical significance for nutrition knowledge, low-fat practices, perceived changes in low-fat practices, and social support. PMID- 11009132 TI - The effect of the Maryland WIC 5-A-Day promotion program on participants' stages of change for fruit and vegetable consumption. AB - The Maryland Women, Infants and Children (WIC) 5-A-Day Promotion Program examined the effect of a multifaceted nutrition intervention on changing the fruit and vegetable consumption of low-income women in the WIC program in Maryland. The sample consisted of 3,122 participants (1,443 intervention and 1,679 control) with a mean age of 27.2. Fifty-six percent were Black/African American. This article focuses on the effect of the intervention on the stages of change of the participants. Intervention participants showed significantly greater positive movement through the stages than control participants. Stages of change were measured for two specific target behaviors: eating five servings of fruits and vegetables a day and eating more servings of fruit and vegetables a day. Results demonstrated significant differences in the stage status of intervention and control women and in movement through the stages. The effectiveness of the intervention across groups depended on which staging measure was used. PMID- 11009131 TI - Dealing with diversity: recruiting churches and women for a randomized trial of mammography promotion. AB - There is little documentation about the recruitment process for church-based health education programs. In this study, the authors recruit African American, Latino, and white churches and women members (age 50 to 80) for a randomized church-based trial of mammography promotion in Los Angeles County. Efforts to enhance recruitment began 10 months before churches were invited to participate and included a variety of community-based strategies. Subsequently, 45 churches were recruited over a 5-month period through group pastor breakfast meetings and church-specific follow-up. In close collaboration with the 45 churches, the authors administered church-based surveys over 6 months and identified 1,967 age eligible women who agreed to be contacted by the program team. It was found that an extended resource intensive period of relationship-building and community based activities were necessary to conduct church-based programs effectively, particularly among older and ethnically diverse urban populations. PMID- 11009133 TI - Enabling research in developing countries. PMID- 11009134 TI - Development aid: manna or myth? PMID- 11009135 TI - Can HIV-1 infections in Africa provide insights into acquired immunity to malaria? PMID- 11009137 TI - More rigour needed in trials of new diagnostic agents for tuberculosis. PMID- 11009136 TI - A 21st century water cure. PMID- 11009138 TI - Xenotransplantation: one trotter forward, one claw back. PMID- 11009139 TI - Effect of HIV-1 and increasing immunosuppression on malaria parasitaemia and clinical episodes in adults in rural Uganda: a cohort study. AB - BACKGROUND: An association between HIV-1 and malaria is expected in theory, but has not been convincingly shown in practice. We studied the effects of HIV-1 infection and advancing immunosuppression on falciparum parasitaemia and clinical malaria. METHODS: HIV-1-positive and HIV-1-negative adults selected from a population-based cohort in rural Uganda were invited to attend a clinic every 3 months (routine visits) and whenever they were sick (interim visits). At each visit, information was collected on recent fever, body temperature, and malaria parasites. Participants were assigned a clinical stage at each routine visit and had regular CD4-cell measurements. FINDINGS: 484 participants made 7220 routine clinic visits between 1990 and 1998. Parasitaemia was more common at visits by HIV-1-positive individuals (328 of 2788 [11.8%] vs 231 of 3688 [6.3%], p<0.0001). At HIV-1-positive visits, lower CD4-cell counts were associated with higher parasite densities, compared with HIV-1-negative visits (p=0.0076). Clinical malaria was significantly more common at HIV-1-positive visits (55 of 2788 [2.0%] vs 26 of 3688 [0.7%], p=0.0003) and the odds of having clinical malaria increased with falling CD4-cell count (p=0.0002) and advancing clinical stage (p=0.0024). Participants made 3377 interim visits. The risk of clinical malaria was significantly higher at visits by HIV-1-positive individuals than HIV-1-negative individuals (4.0% vs 1.9%, p=0.009). The risk of clinical malaria tended to increase with falling CD4-cell counts (p=0.052). INTERPRETATION: HIV-1 infection is associated with an increased frequency of clinical malaria and parasitaemia. This association tends to become more pronounced with advancing immunosuppression, and could have important public-health implications for sub Saharan Africa. PMID- 11009140 TI - Doxycycline and rifampicin for mild scrub-typhus infections in northern Thailand: a randomised trial. AB - BACKGROUND: Some strains of scrub typhus in northern Thailand are poorly responsive to standard antirickettsial drugs. We therefore did a masked, randomised trial to compare rifampicin with standard doxycycline therapy for patients with scrub typhus. METHODS: Adult patients with strictly defined, mild scrub typhus were initially randomly assigned 1 week of daily oral treatment with 200 mg doxycycline (n=40), 600 mg rifampicin (n=38), or doxycycline with rifampicin (n=11). During the first year of treatment, the combined regimen was withdrawn because of lack of efficacy and the regimen was replaced with 900 mg rifampicin (n=37). Treatment outcome was assessed by fever clearance time (the time for oral temperature to fall below 37.3 degrees C). FINDINGS: About 12,800 fever patients were screened during the 3-year study to recruit 126 patients with confirmed scrub typhus and no other infection, of whom 86 completed therapy. Eight individuals received the combined regimen that was discontinued after 1 year. The median duration of pyrexia was significantly shorter (p=0.01) in the 24 patients treated with 900 mg daily rifampicin (fever clearance time 22.5 h) and in the 26 patients who received 600 mg rifampicin (fever clearance time 27.5 h) than in the 28 patients given doxycycline monotherapy (fever clearance time 52 h). Fever resolved in a significantly higher proportion of patients within 48 h of starting rifampicin (900 mg=79% [19 of 24], 600 mg=77% [20 of 26]) than in patients treated with doxycycline (46% [13 of 28]; p=0.02). Severe gastrointestinal events warranted exclusion of two patients on doxycyline. There were two relapses after doxycycline therapy, but none after rifampicin therapy. INTERPRETATION: Rifampicin is more effective than doxycycline against scrub typhus infections acquired in northern Thailand, where strains with reduced susceptibility to antibiotics can occur. PMID- 11009141 TI - Human herpesvirus 8 transmission from mother to child and between siblings in an endemic population. AB - BACKGROUND: Transmission of human herpesvirus 8 (HHV-8), the aetiological agent of Kaposi's sarcoma, is known to occur during sex among homosexual men. However, other modes of HHV-8 transmission remain to be elucidated in endemic populations. METHODS: We did a population-based seroepidemiological survey in a village in French Guiana among 1337 individuals of African origin (age 2-91 years) who had reliable genealogical data. Plasma samples were taken and tested for HHV-specific IgG by immunofluorescence assay. Risk factors and familial correlations for HHV-8 seropositivity were modelled by logistic regression analysis by use of the estimating equations approach, which expresses familial dependences in terms of odds ratios. Familial odds ratios were also acquired by use of the distribution of all possible pairs of a given familial dependence. FINDINGS: The overall HHV-8 seroprevalence was 13.2% with no difference according to sex. HHV-8 seropositivity was strongly age dependent: at 1.2% under 5 years, HHV-8 seroprevalence rose up to a plateau around 15% between 15 and 40 years, and showed a seroprevalence of more than 27% in individuals older than 40 years. Strong familial aggregation in HHV-8 seroprevalence was found with high mother child (odd ratio 2.8 [95% CI 1.6-5.0]) and sib-sib (3.8 [1.6-9.5]) correlations. By contrast, no significant correlation between spouses (0.6 [0.2-1.9]) was seen. INTERPRETATION: This pattern of familial aggregation, together with the variation of HHV-8 seroprevalence with age, indicate that, in endemic populations, HHV-8 transmission mainly occurs from mother to child and between siblings during childhood and adolescence. PMID- 11009143 TI - The Krukenberg procedure: a surgical option for the treatment of double hand amputees in Sierra Leone. AB - BACKGROUND: For the past 9 years, Sierra Leone has struggled with a cruel civil war in which the civilian population has been the principal target. The International Committee of the Red Cross established a surgical mission in Netland Hospital, Freetown, and during this surgical mission, we treated crippled individuals who had had one or both hands amputated. We assessed the benefit of the Krukenberg procedure to restore manual dexterity in this group. In this operation, radius and ulna are separated and a pincer-like grasp is created. METHODS: From July 15, 1998, to Sept 9, 1998, we carried out the Krukenberg procedure on a group of double and single hand amputees. The main target group was double hand amputees, however, the procedure was initially done on three single hand amputees to assess the functional result. Between the original trauma and the operation, there had been an interval of 3 months or longer in all patients. Patients were followed up for 3 months after the procedure to assess gripping strength and self-sufficiency (feeding, personal hygiene, dressing, and manual dexterity). FINDINGS: 15 Krukenberg procedures were done on 11 patients: eight men and three women (mean age 42 years). Three had single hand amputation and eight had double hand amputation. In only two patients had sufficient time passed for the stumps to heal. Of ten patients who were followed up at 3 months, all could eat and drink by themselves. The simple dressing test was passed by all patients and 75% of the patients had recovered some dexterity. INTERPRETATION: The Krukenberg procedure is a surgical option to achieve some manual dexterity in double hand amputees, where resources for sophisticated hand prostheses are lacking. PMID- 11009142 TI - Tuberculosis control and molecular epidemiology in a South African gold-mining community. AB - BACKGROUND: Gold miners have very high rates of tuberculosis. The contribution of infections imported into mining communities versus transmission within them is not known and has implications for control strategies. METHODS: We did a prospective, population-based molecular and conventional epidemiological study of pulmonary tuberculosis in a group of goldminers. Clusters were defined as groups of patients with Mycobacterium tuberculosis isolates with identical IS6110 DNA fingerprints. We compared the frequency of possible risk factors in the clustered and non-clustered patients whose isolates had fingerprints with more than four bands, and re-interviewed members of 45 clusters. FINDINGS: Of 448 patients, ten were excluded because they had false-positive cultures. Fingerprints were made in 419 of 438, of which 371 had more than four bands. 248 of 371 were categorised into 62 clusters. At least 50% of tuberculosis cases were due to transmission within the community. Patients who had failed treatment at entry to the study were more likely to be in clusters (adjusted odds ratio 3.41 [95% CI 1.25-9.27]). Patients with multidrug-resistant isolates were more likely to have failed treatment but were less likely to be clustered than those with a sensitive strain (0.27 [0.09-0.83]). HIV infection was common (177 of 370 tested) but not associated with clustering. INTERPRETATION: Despite a control programme that cures 86% of new cases, most tuberculosis in this mining community is due to ongoing transmission. Persistently infectious individuals who have previously failed treatment may be responsible for one third of tuberculosis cases. WHO targets for cure rates are not sufficient to interrupt transmission of tuberculosis in this setting. Indicators that are more closely linked to the rate of ongoing transmission are needed. PMID- 11009144 TI - Talking to the patient. PMID- 11009145 TI - Satellite mapping of Loa loa prevalence in relation to ivermectin use in west and central Africa. AB - For many years, ivermectin has been widely distributed throughout west Africa for the safe and effective control of onchocerclasis. However, recent events in Loa loa-endemic areas of Cameroon, where severe adverse reactions have occurred, now constrain the public-health use of this drug in the forest habitat of the L. loa vector. We have created a model of L. loa prevalence to identify areas where high endemicity may be associated with the occurrence of such reactions. The model results have been mapped and the areas of overlap between high L. loa prevalence and planned ivermectin distribution for onchocerciasis control identified. PMID- 11009146 TI - Carriage of serogroup W-135, ET-37 meningococci in The Gambia: implications for immunisation policy? AB - We found high levels of symptomless carriage of a hyperinvasive Neisseria meningitidis strain (electrophoretic type 37 [ET-37], serogroup W-135) during a vaccine trial in Gambian children in 1996. Serogroup C, ET-37 complex meningococci cause 30-40% of meningococcal disease in countries such as the UK, and have a point prevalence of 0.5-1.0%. The recent Haj-associated spread of serogroup W-135, ET-37 complex meningococci, which has been accompanied by numerous secondary cases, might be explained by the apparently raised carriage rates reported here. PMID- 11009147 TI - Rickettsia felis rickettsiosis in Yucatan. AB - Three patients with fever, exanthem, headache, and central-nervous-system involvement were diagnosed with Rickettsia fells infection by specific PCR of blood or skin and seroconversion to surrogate Rickettsia antigens. Although R. felis's relationship to other Rickettsia species is known and the pathogenic potential of this clade is well documented, R. felis's role as a pathogen has not been fully understood. PMID- 11009148 TI - Chronic neuropathic pain in treated leprosy. AB - The existence of chronic neuropathic pain in treated leprosy has received scant attention. We describe the clinical findings of 16 patients with multibacillary leprosy who had chronic stimulus-independent pain despite finishing their treatment. With confirmation, our results could be of importance in the establishment of "care after cure" programmes for patients with leprosy. PMID- 11009149 TI - Duration of viraemia and faecal viral excretion in acute hepatitis E. AB - Data on duration of viral excretion and viraemia during hepatitis E virus (HEV) infection are limited. We tested serial stool and serum samples from 20 patients with acute hepatitis E for HEV RNA. Faecal excretion and viraemia in these patients were found to be short lived. In 19 patients, all samples obtained after biochemical resolution of hepatitis tested negative; in the remaining patient, HEV RNA was detected in the serum samples but not in stool after biochemical resolution. Long-term persistence of HEV in body fluids of infected individuals seems to be an unlikely reservoir for transmission of HEV. PMID- 11009151 TI - Assessment of a 1-year teaching programme in Zanzibar, Tanzania. AB - We assessed whether a 1-year teaching programme in northern Zanzibar would improve prescribing practice. Data on polypharmacy and appropriateness of the treatment of upper-respiratory infection, anaemia, and scabies from the 17 primary health-care units in northern Zanzibar were analysed before and after the teaching programme. There was a significant and sustained reduction in polypharmacy and an improvement in the treatment of upper-respiratory infection, scabies, and anaemia. This teaching programme has been successful in improving prescribing practices in a less-developed country. PMID- 11009150 TI - Anthelminthic treatment and haemoglobin concentrations during pregnancy. AB - A longitudinal study was conducted in Sierra Leone to measure the impact of a single dose anthelminthic (400 mg albendazole) and daily iron-folate supplements (36 g iron and 5 mg folate) on haemoglobin (HG) concentration during pregnancy. After controlling for baseline Hb concentration and season, anthelmintic treatment reduced the decline in haemoglobin concentration between the first and third trimesters by 6.6 g/L (p=0.0034) relative to the control. The corresponding value for iron-folate supplements was 13.7 g/L(p<0.0001) [corrected]. These findings indicate that anthelminthic treatment should be included in strategies to control maternal anaemia in Sierra Leone. PMID- 11009152 TI - Women suffer first from lack of health-care services. PMID- 11009153 TI - Single-dose treatment shows effectiveness for Indian visceral leishmaniasis. PMID- 11009154 TI - The "gender gap" in autoimmune disease. PMID- 11009155 TI - India steps up anti-tobacco measures. PMID- 11009156 TI - More confusion about South African President's stance on HIV. PMID- 11009157 TI - Concern about rise in AIDS in Romanian adults. PMID- 11009158 TI - Anger at graphic evidence at Omagh inquest. PMID- 11009159 TI - Explaining trends in inequities: evidence from Brazilian child health studies. AB - There is considerable international concern that child-health inequities seem to be getting worse between and within richer and poorer countries. The "inverse equity hypothesis" is proposed to explain how such health inequities may get worse, remain the same, or improve over time. We postulate that as new public health interventions and programmes initially reach those of higher socioeconomic status and only later affect the poor, there are early increases in inequity ratios for coverage, morbidity, and mortality indicators. Inequities only improve later when the rich have achieved new minimum achievable levels for morbidity and mortality and the poor gain greater access to the interventions. The hypothesis was examined using three epidemiological data sets for time trends in child health inequities within Brazil. Time trends for inequity ratios for morbidity and mortality, which were consistent with the hypothesis, showed both improvements and deterioration over time, despite the indicators showing absolute improvements in health status between rich and poor. PMID- 11009160 TI - Specific immune-based diagnosis of tuberculosis. AB - Current diagnostic tests for tuberculosis based on tuberculin have poor specificity, and both BCG vaccination and exposure to non-tuberculosis mycobacteria produce a response similar to that induced by infection with Mycobacterium tuberculosis. The identification of regions of the M. tuberculosis genome that are not present in BCG and non-tuberculous mycobacteria provides a unique opportunity to develop new highly specific diagnostic reagents. We describe the current status of attempts to exploit this information and summarise recent research that has used defined antigens for an accurate and rapid test for tuberculosis infection based on the detection of T cells sensitised to M. tuberculosis either by blood tests in vitro or skin tests in vivo. PMID- 11009161 TI - Evidence-based medicine--the view from Fiji. PMID- 11009162 TI - Consequences of in-utero death in twin pregnancy. PMID- 11009163 TI - Consequences of in-utero death in twin pregnancy. PMID- 11009164 TI - Cardiovascular disease in South Asians. PMID- 11009165 TI - Testing for West Nile virus. PMID- 11009166 TI - Digital arterial occlusive disease. PMID- 11009167 TI - Intra-arterial thrombolysis for hyperacute stroke. PMID- 11009168 TI - Pumpless extracorporeal lung assist. PMID- 11009169 TI - Breast carcinoma in young patients. PMID- 11009170 TI - Breast carcinoma in young patients. PMID- 11009171 TI - NICE and drugs for multiple sclerosis. National Institute of Clinical Excellence. PMID- 11009172 TI - Jehovah's Witness blood policy. PMID- 11009173 TI - Jehovah's Witness blood policy. PMID- 11009174 TI - Jehovah's Witness blood policy. PMID- 11009175 TI - Ethics of mass STD treatment. PMID- 11009177 TI - On education and training. PMID- 11009176 TI - Ethics of mass STD treatment. PMID- 11009178 TI - Computational models of the basal ganglia. AB - Computer simulation studies and mathematical analysis of models of the basal ganglia are being used increasingly to explore theories of basal ganglia function. We review the implications of these new models for a general understanding of basal ganglia function in normal as well as in diseased brains. The focus is on their functional similarities rather than on the details of mathematical methodologies and simulation techniques. Most of the models suggest a vital role for the basal ganglia in learning. Although this interest in learning is partly driven by experimental results associating the acute firing of dopamine cells with reward prediction in monkeys, some of the models have preceded the electrophysiological results. Another common theme of the models is selection. In this case, the striatum is seen as detecting and selecting cortical contexts for access to basal ganglia output. Although the behavioral consequences of this selection are hard to define, the models provide frameworks within which to explore these ideas empirically. This provides a means of refining our understanding of basal ganglia function and to consider dysfunction within the new logical frameworks. PMID- 11009180 TI - The pathogenesis of multiple system atrophy: past, present, and future. AB - Multiple system atrophy is a sporadic, adult-onset neurodegenerative disease of unknown etiology. The condition may be unique among neurodegenerative diseases by the prominent, if not primary, role played by the oligodendroglial cell in the pathogenetic process. Recent developments in our understanding of multiple system atrophy have included the detection of glial cytoplasmic inclusions and alpha synuclein accumulation in these inclusions. The latter finding links multiple system atrophy as an "alpha-synucleinopathy" to Parkinson's disease and dementia with Lewy bodies. This article reviews recent important findings of potential relevance to the pathogenesis of multiple system atrophy. We also speculate on areas in which further advances may be made to progress our understanding of this devastating condition. PMID- 11009179 TI - Neuropsychologic assessment of patients for movement disorder surgery. AB - The neuropsychologic evaluation of patients under consideration for movement disorder surgery is recognized as being an essential component of the preoperative process. Patients with early-stage concomitant dementia must be identified and the relative risk of postoperative cognitive decline evaluated. Knowledge of the patterns of an individual's strengths and weaknesses might also be a factor in deciding on a neurosurgical procedure. Although the advent of pallidal deep brain stimulation (DBS) has possibly resulted in reduced risk of induced cognitive impairment, even this procedure has been associated with negative sequelae. DBS within the subthalamic nucleus is becoming the method of choice and this may lead to cognitive and behavioral compromise, especially in the elderly patient. The team considering the establishment of neurosurgical treatment is often at a loss to decide how much neuropsychologic testing is required to determine relative risks of cognitive or behavioral morbidity as a consequence of the procedure. A brief summary of expected outcome and of pertinent family process and psychodynamic issues are addressed. This article is intended to serve as a guide to permit clinicians to choose the appropriate length and depth of neuropsychologic assessment, but also to highlight the confounding factors often present in these patients. PMID- 11009181 TI - Apomorphine: an underutilized therapy for Parkinson's disease. AB - Apomorphine was the first dopaminergic drug ever used to treat symptoms of Parkinson's disease. While powerful antiparkinsonian effects had been observed as early as 1951, the potential of treating fluctuating Parkinson's disease by subcutaneous administration of apomorphine has only recently become the subject of systematic study. A number of small scale clinical trials have unequivocally shown that intermittent subcutaneous apomorphine injections produce antiparkinsonian benefit close if not identical to that seen with levodopa and that apomorphine rescue injections can reliably revert off-periods even in patients with complex on-off motor swings. Continuous subcutaneous apomorphine infusions can reduce daily off-time by more than 50% in this group of patients, which appears to be a stronger effect than that generally seen with add-on therapy with oral dopamine agonists or COMT inhibitors. Extended follow-up studies of up to 8 years have demonstrated long-term persistence of apomorphine efficacy. In addition, there is convincing clinical evidence that monotherapy with continuous subcutaneous apomorphine infusions is associated with marked reductions of preexisting levodopa-induced dyskinesias. The main side effects of subcutaneous apomorphine treatment are related to cutaneous tolerability problems, whereas sedation and psychiatric complications play a lesser role. Given the marked degree of efficacy of subcutaneous apomorphine treatment in fluctuating Parkinson's disease, this approach seems to deserve more widespread clinical use. PMID- 11009182 TI - Excitatory responses in the 'direct' striatonigral pathway: effect of nigrostriatal lesion. AB - The existence of a striatonigral GABAergic projection that inhibits substantia nigra reticulata (SNr) cells has been well established. We report on electrophysiological evidence in the rat for a striatonigral excitatory pathway that affects 15% of all SNr cells. Using the antidromic response test to classify SNr cells in relation to their projecting nucleus, an excitatory striatonigral response was found in 57% of cells projecting to the pedunculopontine tegmental nucleus (PPTg) and 11% of cells projecting to the thalamus. SNr cells innervated by inhibitory or excitatory striatonigral inputs had a similar firing rate but a lower variation coefficient (VC) than SNr cells that did not respond to striatonigral inputs. The 6-hydroxydopamine (6-OHDA) degeneration of nigrostriatal dopaminergic cells (A9) induced an increase in the percentage of SNr cells excited from the striatum (52%) and in the VC, but no modification of the firing rate or of the number of spikes induced by each striatal stimulus. This increase in VC was found for the striatonigral inhibitory, but not for the striatonigral excitatory pathway. These data indicate that in addition to its inhibitory action, the direct striatonigral pathway has an excitatory activity that is particularly prominent for SNr cells projecting to the pedunculopontine nucleus. Because the percentage of SNr cells excited by this pathway was markedly increased by DA cell degeneration, our findings suggest that the excitatory striatonigral system could be involved in the pathophysiology of Parkinson's disease. PMID- 11009183 TI - Endogenous dopaminergic tone and dopamine agonist action. AB - Dopamine receptor agonists provide symptomatic relief in the early stages of Parkinson's disease, but with disease progression, their efficacy decreases. The reason behind this decrease in effectiveness is unknown, but maximal efficacy may be dependent on endogenous dopaminergic tone to provide stimulation of D1 and D2 receptor subtypes. Therefore, we have investigated the effects of the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine (AMPT) on the actions of D1, D2, and D1/D2 agonists and levodopa (L-dopa) in common marmosets treated with 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Administration of AMPT alone further increased motor disability and decreased locomotor activity. Administration of L dopa reversed motor disability and increased locomotor activity, and this reversal was not affected by previous AMPT treatment. The D1 agonist A-77636 and the D2 agonist quinpirole reversed motor deficits, but these effects were markedly inhibited by previous AMPT treatment. Administration of quinpirole with A-77636 produced a reversal of motor deficits that was more resistant to AMPT pretreatment than was the effect produced by quinpirole or A-77636 alone. These data suggest that D1 and D2 receptor stimulation are required for dopamine receptor agonists to produce a maximal antiparkinsonian response. The reversal of motor deficits produced by the mixed D1/D2 agonist apomorphine was more resistant to AMPT treatment than that produced by quinpirole or A-77636. However, the motor effects of A-77636 plus quinpirole and of apomorphine were still affected by AMPT treatment. This suggests that loss of tyrosine hydroxylase activity may also alter motor activity through inhibition of endogenous L-dopa or norepinephrine synthesis, because both are also involved in the genesis of motor activity. PMID- 11009184 TI - Alcohol dehydrogenase alleles in Parkinson's disease. AB - Mutations in alcohol dehydrogenase (ADH; EC 1.1.1.1) genes may be of interest in the etiology of Parkinson's disease (PD) because of the important role these enzymes play in retinoid and dopamine metabolism and/or aldehyde detoxification. The location of several alcohol dehydrogenase genes in a cluster on chromosome 4 lends further support to ADH genes being candidates for this disorder, because recently a form of autosomal-dominant parkinsonism has been mapped to this area. We sequenced the promoter and coding regions and part of the introns of the human class IV ADH gene in 10 patients with PD. Seven different polymorphisms were identified. These polymorphisms could be assigned to four alleles (A1-A4). We then determined the frequencies of those four alleles and the wild-type allele in 78 patients with PD and 130 control subjects and found a significant association of the A1 allele with PD (odds ratio = 2.87; 95% confidence interval = 1.35 6.08). In familial cases, the association was strongest (odds ratio = 4.86; 95% confidence interval = 1.89-12.75). Two patients were homozygous for A1 whereas none of the 130 control subjects was found to be homozygous. Our results show an association between a certain ADH4 (formerly known as ADH7 in humans) allele and PD. This suggests a role for genetic variations of ADH4 as risk factors for the development of PD. Our data also show that the observed polymorphisms alone are not sufficient to cause symptoms. Further genetic and/or environmental factors have to be involved. PMID- 11009185 TI - Influence of strict, intermediate, and broad diagnostic criteria on the age- and sex-specific incidence of Parkinson's disease. AB - We studied the influence of three sets of diagnostic criteria on the age- and sex specific incidence of Parkinson's disease (PD) among residents of Olmsted County, Minnesota, for the period 1976 to 1990. Incidence cases of parkinsonism were detected using the medical records-linkage system of the Rochester Epidemiology Project. PD was separated from other types of parkinsonism using strict, intermediate, and broad criteria. We found 154 incident cases of PD using the strict criteria, 215 using the intermediate criteria, and 266 using the broad criteria. The incidence rate was consistently higher for men across all ages with all three sets of criteria; however, sex differences were more striking at older ages when using the broad criteria. In men above age 79 years, the incidence rate of PD declined with strict criteria, remained stable with intermediate criteria, and increased with broad criteria. The impact of diagnostic criteria on the age specific incidence curve was less striking for women. When using the broad criteria, the risk of PD increased constantly with age in both sexes, suggesting that PD is an aging-related disease. Our findings suggest that the diagnostic criteria used to separate PD from other types of parkinsonism influence the magnitude of PD incidence and its distribution by age and sex. PMID- 11009186 TI - Clinical problems in non-fluctuating patients with Parkinson's disease: a community-based study. AB - OBJECTIVE: To investigate the frequency of nonfluctuators in a community-based prevalence study of Parkinson's disease (PD) and to describe disability, non motor problems, and health-related quality of life in patients with PD with and without motor fluctuations, and compare the findings to those of two control groups. METHODS: The study involved 245 patients with PD who were participating in a prevalence study and two control groups (100 healthy elderly individuals and 100 patients with diabetes mellitus [DM]). Data were obtained through neurologic examination and a semistructured interview, and by the use of several questionnaires. RESULTS: In this group of unselected patients with PD, 78% did not experience motor fluctuations. Mean duration of treatment with levodopa was 6.3 years. Patients with motor fluctuations had a lower age at onset of disease, longer duration of disease, and a higher daily levodopa dose than patients without fluctuations. Among the non-fluctuating patients, we found more dementia and a higher age at prevalence day. Disability (assessed by the Unified Parkinson's Disease Rating Scale subscales for activities of daily living and motor function and the Hoehn and Yahr stage) was similar in fluctuators and nonfluctuators. Depression, sleep disturbances, and fatigue were equally frequent in both patient groups. The occurrence of these difficulties was clearly more frequent among non-fluctuating patients with PD than among the control subjects. CONCLUSION: Most patients in the general population who have PD do not experience dose-dependent motor fluctuations. Severity of motor disability and neuropsychiatric manifestations are as important in non-fluctuators as in fluctuators. Patients without motor fluctuations have more depression, sleep disturbances, fatigue, and a poorer health-related quality of life than patients with DM and healthy elderly individuals. This also underlines the importance of developing better management and treatment strategies for this group of patients with PD. PMID- 11009187 TI - Decrease in presynaptic inhibition on heteronymous monosynaptic Ia terminals in patients with Parkinson's disease. AB - PURPOSE: Heteronymous Ia facilitation from the quadriceps to the soleus was studied to clarify central motor control through presynaptic inhibition (PSI) on Ia terminals of spinal motoneurons in Parkinson's disease. METHODS: 17 patients with Parkinson's disease and 36 control subjects participated in the study. Because the early part of facilitation reflects the degree of PSI, the extent of facilitation was quantified as the slope, within 0.8 msec of onset. RESULTS: Heteronymous Ia facilitation in the patients was greater than in the age-matched control participants. PSI was negatively correlated with the walking speed of the patients. In the patients examined twice, lessening of bradykinesia was correlated to a decrease in PSI but not lessening of rigidity. CONCLUSION: These findings indicate that the degree of PSI is decreased and disturbance of the central control of PSI may relate to gait disturbance in Parkinson's disease. PMID- 11009188 TI - Perseveration for novel stimuli in Parkinson's disease: an evaluation based on event-related potentials topography. AB - Event-related potential topography produced by novel and target stimuli was used to detect dysfunction of mental switching (perseveration) in nondemented patients with Parkinson's disease. The study participants were 15 patients with Parkinson's disease and 13 age-matched healthy control patients. Ten percent of the novelty tones with pitches of 125 and 500 Hz were added to 20% of the target tones that had a pitch of 1000 Hz. Patients were instructed to count the target tones. The modified Wisconsin Card Sorting Test was used to evaluate frontal lobe function. Patients with Parkinson's disease showed a significant decrease in the achieved categories and an increase in perseverative errors in the Wisconsin Card Sorting Test. These results indicate that the cognitive impairment of patients with Parkinson's disease can be characterized as failure of mental switching related to frontal lobe dysfunction based on basal ganglia disturbance. As compared with the control patients, patients with Parkinson's disease had shorter P3 latencies to the novel stimuli and a more frontal distribution on the P3 map, especially for the 125-Hz stimuli. This characteristic of P3 to novel stimuli in the patients with Parkinson's disease, but not in the control patients, is categorized by P3a (novelty P3). Our findings suggest that decreased mental switching causes lack of novelty P3 habituation in patients with Parkinson's disease and that it is related to learning disabilities based on dysfunction of the frontal lobe and basal ganglia. PMID- 11009189 TI - Determinants of tapping speed in normal control subjects and subjects with Parkinson's disease: differing effects of brief and continued practice. AB - BACKGROUND: Alternate tapping speed is widely used as a measure of bradykinesia in Parkinson's disease (PD). Tapping speed in normal control subjects and factors that might influence tapping speed have not been systematically examined. OBJECTIVE: To examine the effects of age, hand dominance, and gender on tapping speed in normal control subjects and to compare the effects of practice on tapping speed in normal and PD control subjects. METHODS: Tapping speed for three sequential trials in the dominant and nondominant hand was examined in 100 normal control subjects and 60 subjects with PD. The effect of hourly practice over 26 hours (19 trials) was investigated in 14 normal and 24 PD subjects. RESULTS: The speed with which normal subjects alternately tapped two counters was negatively correlated with age, was greater in the dominant hand, was not related to gender, and improved with short-term practice (three trials) and with continued practice over 26 hours. Parkinsonian subjects, in general, tapped more slowly than normal control subjects and more slowly in the more affected arm. Parkinsonian subjects benefited from short-term practice as much as normal control subjects but, unlike normal control subjects, did not improve with continued practice over 26 hours. CONCLUSIONS: Alternate tapping speed is influenced by age, hand dominance, Parkinson's disease, and practice. Subjects with PD do not benefit as much from continued practice as do normal subjects, suggesting some limitation or impairment of procedural (motor) learning in PD. PMID- 11009190 TI - Lateral leg raising in patients with Parkinson's disease: influence of equilibrium constraint. AB - Patients with Parkinson's disease often have difficulty maintaining postural stability. This impairment is attributed to postural adjustment deficits. We studied the postural adjustments associated with the performance of two complex tasks which differed only in the final equilibrium constraints. Ten patients with Parkinson's disease and six age-matched control subjects were asked to raise one leg laterally to an abduction angle of approximately 45 degrees as fast as possible to the right or left in random order. In the first series of tests, the subjects were instructed to maintain the leg at 45 degrees, whereas in the second series they were instructed to place their foot back on the ground. Recordings included ground reaction forces and kinematics. In the patients with Parkinson's disease the final posture for the first task was never maintained. The strategy used to shift the body weight was different for the two groups. In control subjects, it was initiated by a whole body rotation around the ankle followed by a trunk inclination around the hip. Conversely, in patients with Parkinson's disease, the shift of the body weight was initiated by a trunk inclination around the hip and then by a whole body rotation around the ankle. The amplitude of the trunk inclination toward the supporting side was smaller than in the control subjects. The second task with less severe equilibrium constraints was, on the whole, better performed by the patients even though the same postural adjustment deficits were present. PMID- 11009191 TI - The effect of exercise on pharmacokinetics and pharmacodynamics of levodopa. AB - The aim of our study was to evaluate the influence of low-intensity exercise on levodopa absorption and levodopa motor effect. We studied the pharmacokinetics and pharmacodynamics of levodopa under resting conditions and under a workload of 50 watts for 2 hours on a cycle ergometer in 12 parkinsonian patients with predictable fluctuations of motor disability. The patients attended the hospital on both days in a provoked off state. After a baseline assessment of motor disability using the Columbia University rating scale (CURS scale) and a blood test for measurement of the baseline levodopa concentration in the plasma, 100 mg levodopa and 25 mg benserazide were administered as a single dose orally. Blood samples for measurement of the levodopa concentration in the plasma were taken, and motor assessments were conducted at 15-minute intervals for 240 minutes and at 30-minute intervals from 240 to 360 minutes. All patients were able to perform the exercise program. The baseline Columbia University rating scale score did not differ significantly between both days. The mean levodopa concentration in plasma at half-maximal motor effect tended to be higher during exercise and indicated that the patients needed a higher levodopa concentration in plasma to achieve the half-maximal motor effect. The maximal levodopa concentration in plasma tended to be higher with exercise. Both trends did not reach statistical significance. In summary, there was not a negative or a positive net effect of exercise on pharmacokinetics and pharmacodynamics of levodopa. However, there were two counteracting trends: a trend toward better levodopa absorption and a trend toward a deteriorated concentration-effect correlation. PMID- 11009192 TI - Pathologic gambling in Parkinson's disease: a behavioral manifestation of pharmacologic treatment? AB - We describe 12 patients with Parkinson's disease and pathologic gambling. This association has apparently never been reported. The patients were selected from a Parkinson's disease unit of 250 patients. They met Diagnostic and Statistical Manual of Mental Disorders, 4th edition, criteria for pathologic gambling. All patients underwent a neurologic, psychiatric, and psychologic examination, specifically noting the presence or absence of psychopathology in the spectrum of impulse control disorder and the nature of the gambling. Ten patients started gambling after the onset of Parkinson's disease and treatment with levodopa. The pathologic behavior was exclusively present or was markedly increased in "on" periods in 11 patients. All patients had motor fluctuations at the time of the study. Slot machines were the preferred source of gambling for 10 patients, similar to the Spanish gambling population. That the gambling behavior appears more often in the "on" periods of motor fluctuations and that it begins after the onset of Parkinson's disease in most patients and worsens with levodopa therapy suggest that it could be related to the dopaminergic tone in patients with Parkinson's disease and motor fluctuations (that is, it could represent a behavioral manifestation of pharmacologic treatment). PMID- 11009193 TI - Beneficial effects of amantadine on L-dopa-induced dyskinesias in Parkinson's disease. AB - L-Dopa-induced dyskinesias constitute a challenge to the management of advanced Parkinson's disease. According to recent reports, treatment with the NMDA receptor antagonist amantadine may significantly diminish L-dopa-induced dyskinesias. In the present study, the effect of amantadine on L-dopa-induced dykinesias was assessed in a 5-week, double-blind crossover trial. Dyskinesia severity as assessed following oral L-dopa challenges and by self-scoring dyskinesia diaries were reduced approximately 50% after amantadine treatment compared with baseline or placebo phases. Similarly, dyskinesia assessments on the Unified Parkinson's Disease Rating Scale, part IV (items 32 and 33) also revealed significant improvement after treatment with amantadine. The magnitude of the L-dopa motor response to oral challenges was not different after amantadine or placebo treatment, and there was no significant reduction of daily off-time when patients received active treatment. These results confirm previous observations concerning the antidyskinetic potential of amantadine. PMID- 11009195 TI - An autopsy case of autosomal-recessive juvenile parkinsonism with a homozygous exon 4 deletion in the parkin gene. AB - We report the neuropathologic and genetic features of a 70-year-old man with autosomal-recessive juvenile parkinsonism (AR-JP). At the age of 32 years, he developed a dystonic gait, followed by hand tremor, rigidity, bradykinesia, and impaired postural reflex. Levodopa was effective in ameliorating these symptoms. Pathologic examination of autopsy specimens from this patient revealed loss of pigmented neurons and gliosis in the substantia nigra pars compacta (SNPC), being most pronounced in the medial and ventrolateral regions, and locus ceruleus (LC). The melanin content of the remaining SNPC neurons was low. This feature was less marked in the LC. There were no Lewy bodies, as confirmed by immunostaining for alpha-synuclein. An additional, significant finding in this patient was neuronal loss and fibrillary gliosis in the substantia nigra pars reticulata; this feature has not been reported previously in AR-JP. Gene analysis revealed that this autopsied patient and his siblings had the parkin gene mutation (homozygous exon 4 deletion) that is responsible for the disease. PMID- 11009194 TI - The role of sensory cues in the rehabilitation of parkinsonian patients: a comparison of two physical therapy protocols. AB - We devised a single-blind study to assess the role of providing external sensory cues in the rehabilitation of patients with idiopathic Parkinson's disease (PD). Twenty stable, nondemented patients with PD entered a 6-week rehabilitation program and were randomly assigned to two balanced protocols which were differentiated by the use of external sensory cues ("non-cued" vs "cued"). Patients were evaluated by a neurologist, who was blind to group membership, with the Unified Parkinson's Disease Rating Scale (UPDRS) at baseline, end of treatment, and after 6 weeks. Patient groups were comparable for age, disease duration, and severity. A significant reduction of UPDRS scores (activities of daily living and motor sections) was present after the rehabilitation phase in both groups. However, at follow up, while this clinical improvement had largely faded in the "non-cued" group, mean UPDRS scores of the "cued" group were still significantly lower than baseline values. The incorporation of external sensory cues in the rehabilitation protocol can extend the short-term benefit of physical therapy in moderately disabled patients with PD, possibly as a result of the learning of new motor strategies. "Cued" physical therapy for PD should be targeted to compensate for the defective physiological mechanisms. PMID- 11009196 TI - Deficit of brain and skeletal muscle bioenergetics in progressive supranuclear palsy shown in vivo by phosphorus magnetic resonance spectroscopy. AB - Brain and muscle energy metabolism was assessed in vivo in five patients with progressive supranuclear palsy (PSP) using phosphorous magnetic resonance spectroscopy (31P MRS). 31P MRS disclosed a reduced phosphocreatine (PCr) and an increased calculated free adenosine diphosphate (ADP) in the occipital lobes of all patients. In our patients with PSP, inorganic phosphate (Pi) was significantly increased and Mg2+ was reduced. In the gastrocnemius muscle, Pi at rest was increased in four patients, and the three patients who were able to perform an incremental exercise showed a rate of PCr postexercise recovery slower than control subjects. Our findings show that multisystemic deficit of energy metabolism occurs in PSP and suggest that it may play a role in the pathogenesis of this disorder. PMID- 11009198 TI - Dysarthria and orofacial apraxia in corticobasal degeneration. AB - The authors evaluated dysarthria and orofacial apraxia (OFA) in 10 patients with a clinical diagnosis of corticobasal degeneration (CBD). Nine patients were slightly dysarthric according to the French version of the Frenchay Dysarthria Assessment, which evaluates the motricity of the components of the vocal tract. The severity of dysarthria assessed by an intelligibility score was correlated to the global severity of the disease, but not to the duration of the disease. Voluntary movements of the tongue and the lips were impaired in all patients. OFA, evaluated with simple and sequential gestures, was present in nine patients. Sequential gestures were more frequently impaired. The score of OFA was not correlated to the severity of dysarthria, suggesting independent underlying mechanisms. Thus, when specifically assessed, dysarthria and OFA are more frequent in CBD than usually reported. We propose that the underlying pathophysiology is the result of a deficit in programming and execution of repetitive movements. PMID- 11009197 TI - Voxel-based distribution of metabolic impairment in corticobasal degeneration. AB - This report emphasizes the precise topographic distribution of cerebral metabolic impairment in corticobasal degeneration (CBD) and the pathophysiological differences between CBD and progressive supranuclear palsy (PSP). Statistical parametric mapping (SPM96) analysis of 18FDG positron emission tomography (PET) data was performed in 22 patients with CBD compared with 46 healthy subjects (HS) and 21 patients with PSP who were studied at rest. A statistical threshold of p <0.001 was fixed, further corrected for multiple or independent comparisons (p <0.05). In comparison with HS, the metabolic impairment in CBD was asymmetrically distributed in the putamen, thalamus, precentral (Brodmann's area, BA 4), lateral premotor (BA 6/44) and supplementary motor areas (SMA, BA 6), dorsolateral prefrontal (8/9/46) cortex, and the anterior part of the inferior parietal lobe (BA 40) including the intraparietal sulcus (BA 7/40). A similar hypometabolic pattern was observed for most individual analyses. When PSP was compared with CBD, metabolic impairment predominated in the midbrain, anterior cingulate (BA 24/32), and orbitofrontal regions (BA 10). The reverse contrast showed more posterior involvement in CBD (BA 6 and 5/7/40) including SMA. Our data suggest that multiple components of neural networks related to both movement execution and production of skilled movements are functionally disturbed in CBD compared with both HS and PSP. PMID- 11009199 TI - Thalamic tremor: correlations with three-dimensional magnetic resonance imaging data and pathophysiological mechanisms. AB - Tremor associated with a single focal thalamic lesion has rarely been reported. Furthermore, the exact localization of the lesions is difficult to determine because of the imprecision of "conventional" radiology (computed tomography scan and/or "standard" magnetic resonance imaging). The aim of this study was to identify which thalamic structures are involved in tremor associated with a single focal thalamic lesion. We selected two patients who presented with unilateral postural and kinetic tremor of the upper limb related to a localized thalamic infarction. Three-dimensional T1-weighted magnetic resonance imaging sequence (MP-RAGE sequence) was used to determine the precise topography of the lesions by stereotactic analysis using the atlas of Hassler. The lesions were located within the pulvinar, the sensory nuclei, the mediodorsal nucleus, and the ventral lateral posterior nucleus (according to the classification of Hirai and Jones), the latter including the ventral intermediate nucleus (Vim according to the classification of Hassler). However, the Vim was spared. The subthalamic area, which can induce tremor, was not involved. After having compared the topography of the lesions with the clinical findings, we suggest that thalamic tremors may result from the interruption of the cerebellar outflow tract to the Vim within the thalamus. PMID- 11009200 TI - Irregular jerky tremor, myoclonus, and thalamus: a study using low-frequency stimulation. AB - High-frequency thalamic stimulation alleviates tremor in Parkinson's disease (PD) and essential tremor (ET). The origin of thalamic myoclonus is unexplained and the effects of low-frequency thalamic stimulation on movement control are still unknown. We studied the effects of stimulation at a low frequency of 15 Hz in five drug-free patients (3 PD, 2 ET) 6 months after thalamic implantation of quadripolar electrodes (unilateral in four patients, bilateral in one patient). Clinical, electrophysiological, and videotaped assessment, using a monopolar 15 Hz frequency (3 V, 90 micros) stimulation current applied simultaneously through two adjacent contacts of the electrode, was performed. We observed myoclonus and irregular jerky tremor in the upper limb contralateral to the site of stimulation. The jerks lasted less than 200 ms, were irregular and not synchronous with stimulation, were superimposed on rest or postural tremor, and increased in response to tactile, proprioceptive, or vibratory stimuli. The fact that this complex movement disorder can be induced by low-frequency stimulation in the ventral intermediate nucleus (Vim) of the thalamus suggests that it results, at least partly, from dysfunction of the Vim and possibly adjacent nuclei of the thalamus. PMID- 11009201 TI - Environmental stimulation increases survival in mice transgenic for exon 1 of the Huntington's disease gene. AB - Mice transgenic for the first exon of the human Huntington's disease (HD) gene carrying an expanded CAG repeat expansion (R6/2 line) develop a progressive neurologic phenotype with symptoms resembling those seen in HD. The overt symptoms of R6/2 mice worsen with age, resulting in a rapid decline in health and premature death between 13 and 18 weeks of age. In this study, we characterized the onset and progression of the overt phenotype in R6/2 mice and examined factors that affect the phenotype and life expectancy of these mice. In particular, the effects of altering home cage environment, through changing feeding regimes and providing environmental stimulation, were investigated. We show that changes in feeding regimes significantly improved the general well being and life expectancy of R6/2 mice. Furthermore, we find that various forms of environmental stimulation, including regular behavioral testing, significantly improved the survival of R6/2 mice over and above that resulting from the enhanced feeding regime. The fact that environmental stimulation improves the health and life expectancy in R6/2 mice not only enables the mice to serve as more useful research tools, but also suggests that environmental stimulation may have a beneficial impact on the progression of HD in patients. PMID- 11009202 TI - Spinal and cortical inhibition in Huntington's chorea. AB - In this article we studied spinal and cortical inhibitory mechanisms in patients with Huntington's disease. To evaluate spinal cord inhibitory circuitries, we assessed reciprocal inhibition between antagonist forearm muscles and the recovery cycle of the H reflex in the flexor carpi radialis. Patients showed a significant decrease in the presynaptic phase of reciprocal inhibition reaching a minimum at the conditioning-test interval of 20 msec and an abnormal facilitation of the test H reflex at the conditioning test interval of 40 to 60 msec. Throughout its time course (10-200 msec), the H reflex recovery cycle showed a more prominent facilitation in patients than in control subjects. To assess whether the observed pathophysiological abnormalities might have arisen from an abnormal motor cortical excitability, we examined the recovery cycle of the motor potentials evoked by paired transcranial magnetic stimuli. We found that the inhibitory mechanisms controlling motor cortical excitability were normal. An interpretation of the spinal cord abnormalities is that the intrinsically normal but deafferentated motor cortex in Huntington's disease partly loses its inhibitory control, thus disinhibiting spinal cord circuitry. Our findings from paired transcranial magnetic stimulation suggest that cortical motor areas are not hyperexcitable in Huntington's disease. Hence, the postulated thalamocortical overactivity in experimental models of Huntington's disease needs to be reappraised. PMID- 11009203 TI - Pallido-Luysio-Nigral atrophy revealed by rapidly progressive hemidystonia: a clinical, radiologic, functional, and neuropathologic study. AB - Pallido-luysio-nigral atrophy (PLNA) is a rare neurodegenerative disease in which the clinical and radiologic correlates have not yet been clearly established. A 62-year-old man insidiously developed dystonic postures, choreoathetoid movements, slowness, and stiffness, which initially affected the right hand and foot and progressively spread to the entire right side. T2-weighted magnetic resonance imaging showed increased signal intensity in both left and right medial pallida and in the left substantia nigra. Tests using HMPAO-SPECT and FDG-PET demonstrated left cortical hyperperfusion and hypermetabolism, whereas the left lenticular nucleus was slightly hypometabolic. At age 65, abnormal movements and postures involved all four limbs and the axis causing major gait disturbances, and facial and bulbar muscles atrophied resulting in dysarthria, dysphagia, and impaired breathing. Diffuse amyotrophy and fasciculations also appeared. Death occurred at age 66, 4 years after onset. At autopsy, severe bilateral neuronal loss and gliosis restricted to the pallidum, the subthalamic nucleus, the substantia nigra, and the hypoglossal nucleus were noted, accounting for the diagnosis of PLNA with lower motor neuron involvement. Progressive hemidystonia with adult onset represents an unusual clinical presentation for this disorder. Moreover, this observation indicates that a diagnosis of PLNA should be considered for specific magnetic resonance imaging, SPECT, and/or PET data, and suggests that in PLNA, pallidal dysfunction might play a key role in the dystonic presentation. PMID- 11009205 TI - Quantification of sensory trick impact on tremor amplitude and frequency in 60 patients with head tremor. AB - Head tremor with an obvious head deviation is the typical clinical picture of tremulous cervical dystonia (TCD), whereas head tremor without any significant head deviation allows for the differential diagnosis of dystonic head tremor (DHT) as well as essential head tremor (EHT). Clinical and polyelectromyographic (poly-EMG) studies have shown a suppression of dystonic muscle activity in patients with TCD performing a maneuver called geste antagonistique. The effect of these trick maneuvers on head tremor has not been investigated in patients with DHT and EHT. We studied the impact of sensory trick maneuvers on head tremor amplitude and frequency clinically by using the tremor subscore of the Tsui scale and by means of computer-based accelerometry in 60 patients with head tremor as their major disorder. Based on clinical data (modified Tsui scale: rating of spontaneous head deviation [rotation + lateroflexion + ante-/retroflexion]), pharmacologic response of tremor (propranolol, primidone, or alcohol), family history (postural hand tremor in first-degree relatives), and poly-EMG findings (reciprocal inhibition in neck muscles during voluntary head rotation), 34 patients were diagnosed as having TCD, 14 were classified as having DHT, and 12 patients were diagnosed as having EHT. Using a clinical rating scale, head tremor amplitudes showed a significant decrease compared with baseline during the performance of sensory trick maneuvers in patients with TCD and DHT, but not in patients with EHT. This clinically observed effect was accompanied by a significant reduction in the mean peak power of the dominant frequency in patients with TCD (decrease by 83%, p = 0.0001) and DHT (decrease by 90%, p = 0.01), but not in patients with EHT (decrease by 6%, p = 0.6). Head tremor frequencies showed no significant changes in relation to the trick maneuvers. We conclude that a significant reduction of head tremor amplitude during a sensory trick maneuver is a useful quantitative criterion to distinguish TCD and DHT from EHT. PMID- 11009204 TI - A Yorkshire family with adult-onset cranio-cervical primary torsion dystonia. AB - Although a family history is described in approximately 20% of patients, large families with adult-onset craniocervical primary (idiopathic) torsion dystonia (PTD) are rare. We report a new British family with cranio-cervical dystonia. Seventeen members of the family were examined. Five cases were diagnosed as definite PTD and one as probable PTD. Mean age at onset was 29 years (range, 19 40 yrs). The phenotype was characterized by adult-onset cranio-cervical dystonia in all affected cases. A few cases had additional voice tremor and/or postural arm tremor. The GAG deletion in the DYT1 gene was excluded in the index case. Linkage analysis was performed between the disease and several marker loci spanning DYT6 and DYT7 regions, and haplotypes were reconstructed in all subjects. Although linkage analysis was not completely informative, reconstructed haplotypes excluded linkage between the disease and either DYT6 or DYT7. This report confirms that familial cranio-cervical dystonia is genetically heterogeneous, and further studies of other PTD families with similar clinical features are needed to identify other new genes. PMID- 11009206 TI - Disturbed sensorimotor processing during control of precision grip in patients with writer's cramp. AB - The aim of the study was to investigate force regulation in patients with writer's cramp when performing a drawer-opening task using the precision grip. Experimental conditions included intervening load pulses and vibratory manipulations for examining grip force responses to sensory disturbances. The data revealed that grip force was increased in patients with writer's cramp compared with normal subjects, with a stronger modulation in the symptomatic compared with the asymptomatic hand. This denotes a change in force scaling capabilities and most notably for the preferred hand used in manipulative activities. Vibratory stimulation of the extrinsic hand/finger muscles resulted in an increased grip force of both hands in the patients with writer's cramp. The latter was not observed in normal subjects and supports a bilateral dysfunction in sensorimotor integration resulting from focal dystonia. In conclusion, the disturbed regulation of the precision grip during a drawer-opening task is illustrative for the inability of patients with writer's cramp to efficiently control the force output during manipulative activities. PMID- 11009207 TI - Botulinum toxin antibody testing: comparison between the mouse protection assay and the mouse lethality assay. AB - Conventionally, the standard test for detection of antibodies against botulinum toxin (BT-A) has been the mouse lethality assay (MLA). Because this test has a number of disadvantages, a novel mouse protection assay (MPA) was recently introduced. We sought to compare the results of both tests. Forty-three samples from 38 patients with cervical dystonia and complete or partial subjective BT-A therapy failure underwent simultaneous MPA and MLA testing. Twenty-seven samples showed concordant results in both tests. Eleven of them were MPA- and MLA positive and 16 MPA- and MLA-negative, resulting in a significant association of the dichotomous test results (Fisher exact test, p <0.01). Sixteen samples showed discordant results. All of those were MPA-positive and MLA-negative. This excess of MPA-positive results was also significant (Wilcoxon signed-rank test, p <0.001). Of the patients with MPA-positive samples, 62% had complete and 38% had partial therapy failure. Of the patients with MLA-positive samples, 90% had complete and 10% had partial therapy failure. MPA and MLA results show significant association. Statistical analysis and predominance of partial therapy failure in MPA-positive patients demonstrate higher sensitivity of MPA. With its methodologic advantages, its test parameter being more relevant to BT-A therapy, and its higher sensitivity, the MPA appears to be superior to the MLA. PMID- 11009208 TI - Isolation and 13C-NMR characterization of an insoluble proteinaceous fraction from substantia nigra of patients with Parkinson's disease. AB - Neuromelanin is a dark brown pigment suspected of being involved in the pathogenesis of Parkinson's disease. This pigment can be isolated from normal human substantia nigra by a procedure that includes an extensive proteolytic treatment. In this study we used such a procedure to extract the neuromelanin pigment from a pool of substantia nigra from patients affected by Parkinson's disease. 13C Cross polarization magic angle spinning nuclear magnetic resonance spectroscopy and electron paramagnetic resonance spectroscopy were used to characterize the solid residue obtained from the extraction procedure. We found that the pigment extracted from the substantia nigra of parkinsonian patients was mainly composed of highly cross-linked, protease-resistant, lipo-proteic material, whereas the neuromelanin macromolecule appears to be only a minor component of this extract. A synthetic model of melanoprotein has been prepared by enzymatic oxidation of dopamine in the presence of albumin. Once it has undergone the same proteolytic treatment, this model system yields a 13C-NMR spectrum which is similar to that observed for the parkinsonian midbrain extract. These results are consistent with the view that oxidative stress has a relevant role in the pathogenesis of Parkinson's disease. PMID- 11009209 TI - Increased blink rate in advanced Parkinson's disease: a form of 'off'-period dystonia? AB - Blink rates were measured in 25 patients with fluctuating Parkinson's disease in the "off' and "on" periods. In 17 patients, the "off"-period blink rate was low and increased after administration of levodopa. In the remaining eight patients, the "off"-period blink rate was high and returned to normal levels after levodopa. It is suggested that increased blink rate in the latter group represents a new form of "off"-period dystonia. PMID- 11009210 TI - Tolerability of paroxetine in Parkinson's disease: a prospective study. AB - Depression is a common finding in patients with Parkinson's disease (PD). Traditionally, depression has been treated with tricyclic antidepressants, which are often associated with undesirable side effects that may limit their use in PD. Few studies have been performed with selective serotonin reuptake inhibitors (SSRIs) in these patients. We assessed the tolerability of the SSRI antidepressant paroxetine (10-20 mg once per day) in 65 outpatients with PD and depression for a period of at least 3 months. Treatment was continued for 125.3+/ 89.6 days (mean +/- standard deviation) in 52 patients. In these subjects the Hamilton Disease Rating Scale improved from 21.7+/-6.4 to 13.8+/-5.8 (p <0.001). Overall, 13 patients stopped paroxetine after 9.6+/-10.6 days because of adverse reactions. Two patients reported increased "off" time and tremor that reversed after treatment was stopped. No risk factors for intolerance were identified. Paroxetine is a safe and effective drug to treat depression in PD. PMID- 11009211 TI - Neuronal intranuclear inclusion disease and juvenile parkinsonism. AB - Juvenile parkinsonism (onset age <20 yrs) is uncommon and few cases with neuropathologic confirmation have been reported. We present the case of a 17-year old boy who presented with asymmetric arm tremor and bulbar symptoms. His paternal great aunt had parkinsonism with onset at age 22 years. Examination revealed parkinsonism in the absence of additional neurologic signs except for delayed pupillary responses to light. He responded well to levodopa but developed motor fluctuations and disabling dyskinesias after 3 years of treatment. Following attempted withdrawal of levodopa at age 24 years, he developed severe aspiration pneumonia complicated by cardiorepiratory arrests and he died 6 months later. At autopsy, the dominant histologic feature was wide-spread neuronal hyaline intranuclear inclusions. Neuronal depletion was observed in the substantia nigra, locus ceruleus, and, to a lesser extent, in the frontal cortex, and inclusions were particularly prominent in these areas. Inclusions were immunoreactive for ubiquitin and were typical of those seen in neuronal intranuclear inclusion disease (NIID), a rare, multisytem neurodegenerative disease. NIID should be considered in the differential diagnosis of juvenile parkinsonism. A link between NIID and hereditary neurodegenerative disorders characterized by expanded polyglutamine tracts is supported by the similar appearance of intranuclear inclusions in both conditions and by a family history in some cases of NIID. PMID- 11009212 TI - The first identified French family with dentatorubral-pallidoluysian atrophy. AB - We report the first French family with dentatorubral-pallidoluysian atrophy (DRPLA) in which three members, a 36-year-old woman (proband), her 34-year-old sister, and 14-year-old brother were affected. There was no family history of DRPLA and their father presented at age 66 with pes cavus but without any other neurologic symptoms. Molecular analysis of the DRPLA gene from blood leukocytes showed CAG repeat sizes to be 68/16 in the proband, 62/15 in her father, and 16/16 in her mother. This study provides support for the variable clinical presentation of this disease with incomplete penetrance in the father and demonstrates that DRPLA can be observed in the French Caucasian population. PMID- 11009213 TI - Exploration of motor cortex excitability in a diabetic patient with hemiballism hemichorea. AB - Hemiballism-hemichorea in older patients with hyperglycemia, associated with high signal intensity in the contralateral striatum on T1-weighted magnetic resonance scans, is now an accepted clinical entity. We present an additional patient with this disorder. Using transcranial magnetic stimulation, we show that intracortical inhibition in the motor cortex contralateral to hemiballism hemichorea is increased. This finding is discussed in the context of current models of basal ganglia-thalamo-cortical connectivity. PMID- 11009214 TI - Progressive multifocal leukoencephalopathy presenting with an isolated focal movement disorder. AB - Progressive multifocal leukoencephalopathy (PML) is a rare but fatal papovavirus infection of the central nervous system predominantly affecting immunocompromised patients. Although the basal ganglia circuitry may be involved in the pathology of PML, movement disorders are exceedingly rare as presenting symptoms and have not been described as isolated features in such patients. We report a previously healthy, immunocompetent 24-year-old woman with histologically proven PML who presented with a focal movement disorder of the left arm as an isolated symptom for many months before diagnosis. PMID- 11009216 TI - Meta-analysis of polymorphism of the catechol-O-methyltransferase gene in relation to the etiology of Parkinson's disease in Japan. PMID- 11009215 TI - Life-threatening dystonia-dyskinesias in a child: successful treatment with bilateral pallidal stimulation. AB - We report a 13-year-old boy who developed severe, refractory dystonia-dyskinesias as an abrupt worsening of a previously nonprogressive movement disorder. The movements became continuous, requiring artificial respiration and continuous sedation in the intensive-care unit. Various drugs and drug combinations failed to achieve control. The child was then treated successfully with bilateral pallidal (GPi) stimulation as shown in the videotape. Four months later and without medication, the boy regained autonomous gait and audible speech; his neurologic condition continues to improve. PMID- 11009217 TI - Is there addiction to levodopa in patients with Parkinson's disease? PMID- 11009218 TI - The use of NMDA antagonist memantine in drug-resistant dyskinesias resulting from L-dopa. PMID- 11009219 TI - Cerebrotendinous xanthomatosis with predominant parkinsonian syndrome: further confirmation of the clinical heterogeneity. PMID- 11009220 TI - Clinical and genetic study of familial essential tremor in an isolate of Northern Tajikistan. PMID- 11009221 TI - Bilateral striatal necrosis associated with Mycoplasma pneumoniae infection in an adolescent: clinical and neuroradiologic follow up. PMID- 11009222 TI - Jaw-opening dystonia presumably caused by a pontine lesion. PMID- 11009223 TI - Eyelid dystonia in Machado-Joseph disease. PMID- 11009224 TI - Choreic movements induced by cibenzoline: an Ic class antiarrhythmic effect? PMID- 11009225 TI - Parkinsonism in HIV encephalopathy. PMID- 11009226 TI - Reversing tissue injury-induced plastic changes in the spinal cord: the search for the magic bullet. PMID- 11009227 TI - Introduction: Gaston Labat Award 2000--Gary R. Strichartz, Ph.D. PMID- 11009228 TI - Pathways and obstacles to local anesthesia, a personal account: the 2000 Gaston Labat lecture. PMID- 11009229 TI - Effect of oral ketamine on secondary hyperalgesia, thermal and mechanical pain thresholds, and sedation in humans. AB - BACKGROUND AND OBJECTIVES: Ketamine is an N-methyl-D-aspartate (NMDA) receptor antagonist, and has been proven effective in alleviating secondary hyperalgesia in human subjects when injected intravenously. After oral ingestion, ketamine is metabolized into norketamine, which in vitro possesses NMDA receptor antagonistic effect. The aim of this study was to investigate the effects of oral administration of ketamine on secondary hyperalgesia evoked by standardized tissue injury. METHODS: Twenty-four male volunteers were included in the study. Each volunteer received the following treatment regimen, in randomized, double blind, 3-way cross-over fashion: (A) placebo; (B) ketamine, 0.5 mg/kg; and (C) ketamine, 1.0 mg/kg. Standardized tissue injury was induced after study medication by heating the right calf with a rectangular thermode. The temperature was 47 degrees C, and heating time was 7 minutes. The following parameters were investigated: Pain during induction of the burn injury; heat-pain detection thresholds in the injured area and a corresponding noninjured area; secondary hyperalgesia surrounding the injured area on the calf; secondary hyperalgesia induced by heating an area on the thigh with 45 degrees C in 3 minutes; pressure pain detection thresholds measured on the middle phalanx of the 4th left finger; pain during a 60-second thermal stimulation of 46 degrees C on undamaged skin on the left thigh; and side effects. RESULTS: Some degree of sedation was observed after oral administration of ketamine. No effects on any of the other investigated parameters were observed. CONCLUSION: Oral ketamine 0.5 or 1.0 mg/kg has no effect on secondary hyperalgesia or thermal or mechanical pain thresholds in human volunteers. PMID- 11009230 TI - Efficacy of oral mexiletine for neuropathic pain with allodynia: a double-blind, placebo-controlled, crossover study. AB - BACKGROUND AND OBJECTIVES: Mexiletine is an oral sodium channel antagonist that has been reported to be effective in a variety of neuropathic pain syndromes. However, recent reports question the efficacy of oral mexiletine in neuropathic pain. The objectives of this study were to examine the effect of oral mexiletine on pain, neurosensation, allodynia, and quality of life. METHODS: Twenty subjects suffering from neuropathic pain with prominent allodynia were enrolled in a randomized placebo-controlled crossover study. Patients were titrated to a maximum dose of 900 mg/d or dose-limiting side effects, whichever occurred first. At baseline and on days 0, 4, 7, and 10, the following tests were performed: (1) Quality of Life Questionnaires; (2) pain scores; (3) area of allodynia; (4) side effects; (5) neurosensory testing; and (6) peak and trough plasma mexiletine levels. RESULTS: Peak plasma levels occurred on day 10 and were 0.54 microg/mL. There was no significant effect on any quality of life measurement. There was no significant effect on any neurosensory threshold or the area of allodynia. There was a significant effect of mexiletine on stroking-induced pain. There were no significant effects on any other pain score. Side effects were negligible. CONCLUSIONS: At doses of up to 900 mg/d, mexiletine has minimal effects on pain and allodynia of neuropathic pain. However, side effects may preclude higher doses. PMID- 11009231 TI - Effect of oral mexiletine on capsaicin-induced allodynia and hyperalgesia: a double-blind, placebo-controlled, crossover study. AB - BACKGROUND AND OBJECTIVES: Mexiletine is a sodium channel blocker that has been used for the treatment of a variety of neuropathic pain syndromes. A recent double-blinded placebo-controlled study concluded that it was ineffective in the treatment of allodynia associated with neuropathic pain. However, this study failed to achieve adequate plasma levels of mexiletine. This was a study in healthy volunteers that sought to push the drug to dose-limiting side effects and then evaluate the effects on human experimental pain. METHODS: Twelve healthy volunteers were studied using a randomized, double-blind, placebo-controlled crossover study. The subjects were titrated to a maximum dose of 1,350 mg/d or dose-limiting side effects, whichever occurred first. At baseline and day 10 and 17, neurosensory testing, train-of-three thermal pulses, and side-effect measurements were performed and on day 17, intradermal capsaicin was injected on the volar aspect of the forearm and the pain and secondary hyperalgesia to von Frey hair, stroking, and thermal stimuli were measured. RESULTS: Peak plasma levels occurred on day 10 and were 0.36 +/- 0.21 microg/mL. All subjects experienced dose-limiting side effects. The mean maximum tolerable daily dose achieved was 859 mg (range, 300 to 1,350 mg). The side effects reported by the subjects included nausea, lightheadedness, muscle twitching and weakness, blurred vision, headache, tremors, difficulty concentrating, dysphoria, sedation, pruritus, and rash. These side effects occurred at an average daily dose of 993 mg (range, 600 to 1,350 mg). Compared with placebo, mexiletine had no significant effects on any of the neurosensory thresholds and pain scores after intradermal capsaicin. There was a significant reduction in the area of secondary hyperalgesia to von Frey hair stimulation only. There was a significant correlation between plasma mexiletine level and flare response. CONCLUSIONS: Mexiletine has minimal effects on human experimental pain. It is severely limited by side effects and tolerable doses seem to be void of effects on normal neurosensation and facilitated pain induced by capsaicin and thermal heat pulses. PMID- 11009232 TI - Current perception thresholds and postoperative pain in schizophrenic patients. AB - BACKGROUND AND OBJECTIVES: Schizophrenic patients may have less sensitivity to pain; however, pain insensitivity in schizophrenia has not been adequately evaluated. We investigated current perception threshold (CPT) and postoperative pain intensity in patients with long-standing and treated schizophrenia and control patients. METHODS: We measured CPTs for 2,000 Hz, 250 Hz, and 5 Hz and postoperative pain intensity using a visual analogue scale (VAS) in 50 chronic schizophrenic patients who were on chronic phenothiazine derivatives (> 10 years) and for 25 control patients. RESULTS: CPTs for 2,000 Hz, 250 Hz, and 5 Hz in schizophrenic patients were 334.2 +/- 112.2, 303.9 +/- 117.1, and 165.0 +/- 72.3, respectively. CPTs for 2,000 Hz, 250 Hz, and 5 Hz in schizophrenic patients were significantly higher than those of control patients. VAS pain scores of schizophrenic patients were 4.0 +/- 1.7 at 2 hours post-operatively and 3.8 +/- 1.5 at 5 hours postoperatively, which were significantly (P < .05) lower than those (5.0 +/- 1.6 and 5.1 +/- 1.9) of the control group. CONCLUSIONS: Chronic schizophrenic patients have increased current perception threshold and lower VAS pain scores in postoperative pain compared with control patients. PMID- 11009233 TI - Comparison of 0.5% intrathecal bupivacaine with 0.5% intrathecal ropivacaine in the treatment of refractory cancer and noncancer pain conditions: results from a prospective, crossover, double-blind, randomized study. AB - BACKGROUND AND OBJECTIVES: Intrathecal (IT) administration of bupivacaine (BUP) for treatment of "refractory" pain has sometimes been associated with unacceptable side effects. This study was undertaken to determine if IT ropivacaine (ROP) can reduce the rate and intensity of these side effects e.g., urinary retention, paresthesia, and particularly, paresis with gait impairment. A prospective, crossover, double-blind, randomized study. METHODS: Twenty-one patients were enrolled, 9 dropped out of the study, and data were analyzed from 12 patients. Patients were treated by insertion of IT tunneled nylon catheters, continuous infusion of 0.5% ROP followed by 0.5% BUP or 0.5% BUP followed by 0.5% ROP solutions from an external electronic pump. Each local anesthetic was infused for 7 days, and their order of infusion randomized. The comparative efficacy of the ROP and BUP IT infusions was assessed from the daily doses of IT ROP and IT BUP, oral and parenteral opioids, and daily scores of nonopioid analgetic and sedative drug consumption. Self-reported pain intensity (visual analogue scale [VAS] mean scores) and scores of Bromage relaxation, ambulation, nocturnal sleep pattern, rates of side-effects attributable to the IT drugs, the patients' assessment of the IT ROP v the IT BUP periods of the trial, and the comparative daily cost of IT ROP v IT BUP were recorded. RESULTS: The daily doses of the local anesthetics used were 23% higher for ROP than for BUP. Further, the daily cost was approximately equals 3 times higher for ROP than for BUP. No other significant differences between IT ROP and IT BUP were found. CONCLUSION: The results of this study do not support the hypothesis that IT infusion of 0.5% ROP has advantages over IT infusion of 0.5% BUP when administered for relief of "refractory" pain. PMID- 11009234 TI - Technical aspects and postoperative sequelae of spinal and epidural anesthesia: a prospective study of 3,230 orthopedic patients. AB - BACKGROUND AND OBJECTIVES: Major complications after spinal or epidural anesthesia are extremely rare. The occurrence of less serious and transient sequelae and complaints may be underestimated if there is no established organization for the systematic and continuous surveillance of patients after anesthesia. This study was designed to evaluate the possible relationship between various block-related occurrences and the intra- and postoperative side effects and complaints. METHODS: This prospective study included 3,230 orthopedic patients operated on under neuraxial block. The block was performed by single dose (single-shot spinal anesthesia [SPIN], 80.6%), continuous spinal anesthesia (CSA, 10.3%), or combined spinal and epidural anesthesia (CSE, 9.1%) technique. The patient position during surgery and a detailed description of block performance and equipment, as well as all intraoperative problems, were immediately recorded. Every patient was given a standardized questionnaire to be completed and returned after 1 week. RESULTS: The overall incidence of paresthesia was 12.8%, being most frequent during spinal catheter insertion with multiple attempts. Postoperative sensory disturbances (numbness, dysesthesia) occurred unrelated to paresthesia elicited by the puncture or catheterization. The failure rates (SPIN, 1.0%; CSA, 1.5%; and CSE, 1.0%) were quite low. Asystole with successful resuscitation, occurred in 1 patient with CSE but, overall, hemodynamic perturbations were more common with the continuous techniques. On the first postoperative day, decreased sensation of the skin was significantly more frequent after the continuous techniques, still present in 6.5% of SPIN patients, 6.1% of CSA patients, and 17.2% of CSE patients 1 week after surgery. These disturbances were also related to the use of a tourniquet and lateral position of the patient during surgery. The frequency of postdural puncture headache (PDPH) was similar for the different techniques (SPIN, 0.9%; CSA, 1.5%; and CSE, 1.7%). About one third of the patients reported strong postoperative pain on the day after the operation, and 5.6% continued to report this at 1 week. CONCLUSIONS: Although sensory changes were quite frequent, they were mild and transient. Rather than having been caused by the anesthetic technique, per se, a "nonanesthetic" reason (position, tourniquet, immobilization) should also be considered as their origin. Sensory disturbances, as well as strong pain for at least a week, were reported by several patients, and to become aware of these problems and improve the quality of treatment, a universal regional anesthesia follow-up system is recommended. PMID- 11009235 TI - Is urinary drainage necessary during continuous epidural analgesia after colonic resection? AB - BACKGROUND AND OBJECTIVES: Postoperative urinary retention may occur in between 10% and 60% of patients after major surgery. Continuous lumbar epidural analgesia, in contrast to thoracic epidural analgesia, may inhibit urinary bladder function. Postoperative urinary drainage has been common in patients with continuous epidural analgesia, despite the lack of scientific evidence for its indication after thoracic epidural analgesia. This study describes 100 patients who underwent elective colonic resection with 48 hours of continuous thoracic epidural analgesia and only 24 hours of urinary drainage. METHODS: This is a prospective, uncontrolled study with well-defined general anesthesia, postoperative analgesia, and nursing care programs in patients with a planned 2 day hospital stay, urinary catheter removal on the first postoperative morning, and epidural catheter removal on the second postoperative morning. Follow-up in the outpatient clinic was on days 8 and 30. RESULTS: Nine patients needed bladder recatheterization, 8 as a single procedure and 1 patient a second recatheterization with removal on day 7. This patient had urinary infection on day 10 and was readmitted for 5 days because of urosepsis and, subsequently, for cystitis and left-sided epididymitis. Three patients had uncomplicated urinary infection. No patients had urological complaints at 30 days follow-up (95% confidence limit, 0% to 3.6%). CONCLUSION: The low incidence of urinary retention (9%) and urinary infection (4%) suggests that routine bladder catheterization beyond postoperative day 1 may not be necessary in patients with ongoing continuous low-dose thoracic epidural analgesia. PMID- 11009236 TI - Effect of clonidine on upper extremity tourniquet pain in healthy volunteers. AB - BACKGROUND AND OBJECTIVES: Tourniquet pain is often a limiting factor during intravenous regional anesthesia (IVRA). The purpose of this study was to evaluate the efficacy of 1 microg/kg of clonidine added to IVRA-lidocaine in decreasing the onset of severe tourniquet pain. METHODS: Upper extremity IVRA was performed in 15 volunteers with a double-cuffed tourniquet on 2 separate occasions at least 1 week apart. IVRA was established with either 40 mL 0.5% lidocaine (IVRA-L) or 40 mL 0.5% lidocaine with 1 microg/kg clonidine (IVRA-Cl). Verbal pain scores (VPS) from 0 to 10 were recorded every 5 minutes. When the VPS reached 6, the distal cuff was inflated, and the proximal cuff was deflated. This was defined as the first tourniquet time (T1). The study was terminated at a VPS of 10, or at 60 minutes, whichever occurred first. The time from distal cuff inflation to deflation was defined as the second tourniquet time (T2). Total tourniquet time (TT) was the sum of T1 and T2. RESULTS: T1 for IVRA-L (21.6 +/- 3.9) and IVRA-Cl (22.7 +/- 2.7) were not significantly different. T2 and TT were significantly longer (P < .0001; P < .0007, respectively) for IVRA-Cl (33.0 +/- 6.2; 55.6 +/- 6.6) than for IVRA-L (25.5 +/- 4.4; 47.1 +/- 5.2). CONCLUSION: This study shows that the addition of 1 microg/kg of clonidine to 40 mL of 0.5% IVRA-L delays the onset time of tourniquet pain in healthy, unsedated volunteers. PMID- 11009237 TI - Postoperative analgesia after peripheral nerve block for podiatric surgery: clinical efficacy and chemical stability of lidocaine alone versus lidocaine plus ketorolac. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to determine whether the addition of ketorolac tromethamine to local anesthesia for ankle block alters the quality or duration of analgesia after podiatric surgery. The second aim was to determine the chemical stability of ketorolac tromethamine when added to local anesthetic solutions. METHODS: The study design was double-blinded, placebo controlled, and randomized. Seventy-nine American Society of Anesthesiologists (ASA) class I or II patients scheduled for bunionectomy or hammer toe repair, or both were randomized to 1 of 4 groups. Group L received plain 1.73% lidocaine for their ankle block. Group K received 1.73% lidocaine with ketorolac (4 mg/mL) added to the local solution. Group Kiv received 1.73% plain lidocaine for ankle block and 20 mg of ketorolac intravenously. Group E received 1.73% lidocaine with .67% ethanol added. The final concentration of lidocaine for all groups was 1.73%. The block performed in each patient was a 5-point ankle block. Beginning at 1 hour after the completion of the block and every 30 minutes thereafter, visual analogue scale (VAS) and verbal pain scores were recorded. The time from performance of the block to the initial pain and time to the first oral pain medication intake were also recorded. The time and amount of postoperative oral analgesics in the first 9 hours after the block were recorded. Adverse events were also recorded for each group. RESULTS: There were significantly lower overall VAS and verbal pain scores for group K compared with groups E and L and group Kiv compared with group E. Group K also had a significantly longer time to the first reported pain and first oral pain medications than groups E and L, but not with Group Kiv. The same group had significantly fewer average doses of pain medications postoperatively than Groups E and L. Group E had significantly shorter times to first report of pain and first pain medications and higher mean dose of postoperative oral analgesics than group K and Group Kiv. There were no untoward side effects reported from any group. Chemical analysis by gas chromatography (GC) and capillary electrophoresis (CE) showed no significant change in composition of the solutions when ketorolac was mixed with lidocaine and/or bupivacaine and stored at 37 degrees C for 1 week. CONCLUSIONS: The addition of ketorolac to lidocaine for ankle block contributed to longer duration and better quality analgesia after foot surgery compared with plain 1.73% lidocaine or 1.73% lidocaine plus intravenous ketorolac. The ethanol vehicle is unlikely responsible for the analgesic effects of ketorolac. Ketorolac retains its chemical stability when placed in local solutions of lidocaine or bupivacaine. PMID- 11009238 TI - Alkalinized lidocaine and bupivacaine with hyaluronidase for sub-tenon's ophthalmic block. AB - BACKGROUND AND OBJECTIVES: Alkalinization of local anesthetics has been shown to decrease the onset and prolong the duration of block for extraconal and intraconal application in ocular surgery. The objective of this study is to determine if alkalinization is also effective in sub-Tenon's block when hyaluronidase is added to the drug mixture. METHODS: Twenty-nine patients were randomly assigned to 2 groups in a double-blind, prospective fashion to receive 5.125 mL of either a plain mixture LBH (2.5 mL lidocaine 2%, 2.5 mL bupivacaine 0.5%, 5 IU/mL hyaluronidase, and 0.125 mL isotonic saline) or pH-adjusted mixture LBH-PH (2.5 mL lidocaine 2%, 2.5 mL bupivacaine 0.5%, 5 IU/mL hyaluronidase, and 0.125 mL sodium bicarbonate 8.4%) of local anesthetics in a 1-quadrant sub Tenon's block. Time to onset and time to full akinesia were determined every 30 seconds. RESULTS: No difference was found between the study groups. CONCLUSION: pH adjustment of the local anesthetic mixture of lidocaine, bupivacaine, and hyaluronidase offered no additional benefit in sub-Tenon's technique in ocular procedures. PMID- 11009239 TI - Alkalinization and precipitation characteristics of 0.2% ropivacaine. AB - BACKGROUND AND OBJECTIVES: Alkalinization of local anesthetics has been used to increase the speed of onset of nerve blocks. However, alkalinization of local anesthetic solutions may cause precipitation, thereby decreasing bioavailability and anesthetic activity. Alkalinization of ropivacaine has not been described. This laboratory study assessed the alkalinization and precipitation characteristics of ropivacaine. METHODS: Aliquots (2 mL) of commercially available ropivacaine (Naropin, 0.2%; Astra Pharmaceutical, Westborough, MA) were alkalinized with increasing amounts (0.01, 0.02, 0.04 mL) of sodium bicarbonate (8.4%) and immediately monitored for pH change and onset of visible precipitation at room temperature. We then alkalinized ropivacaine with sodium bicarbonate and measured the amount of precipitate that accumulated after various incubation times. RESULTS: The pH of ropivacaine increases with the addition of small amounts of bicarbonate. The calculated percentage of nonionized ropivacaine increased from 0.3% to greater than 30% with alkalinization from pH of 5.51 to 7.63. Drug loss to precipitation increased with higher doses of bicarbonate, reaching 25% to 30% of the total ropivacaine. Even with a low dose of bicarbonate (0.1 mL bicarbonate/20 mL ropivacaine), precipitation increased with time of incubation, reaching a plateau at 20 minutes. CONCLUSIONS: A laboratory evaluation that establishes the alkalinization characteristics of ropivacaine is a prerequisite for designing a clinical study of alkalinized ropivacaine. In our experiment, low doses of bicarbonate produced significant increases in the proportion of nonionized ropivacaine with only modest precipitation. There would be a low likelihood of substantial drug precipitation if the mixture was administered within 5 to 10 minutes after alkalinization. These results indicate that alkalinized ropivacaine should not be used for infusions and that ropivacaine should not be alkalinized until just before use. PMID- 11009240 TI - Antinociceptive and motor-blocking action of epidurally administered IQB-9302 and bupivacaine in the dog. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to compare the antinociceptive and motor-blocking effects of epidurally administered IQB-9302 (C18H26N2O.HCl) and bupivacaine in the dog. METHODS: Twelve adult female Beagle dogs were used. Each animal received 3 concentrations (0.25%, 0.50%, and 0.75%) of either IQB-9302 (n = 6) or bupivacaine (n = 6) by means of a chronic epidural catheter. The nocifensive and motor-blocking status were determined at regular intervals before (baseline) and after drug administration. RESULTS: Epidurally administered IQB-9302 caused a more potent nocifensive and motor-blocking action than bupivacaine. The duration of complete nocifensive block was the longest with IQB-9302, whereas the duration of dermatome nocifensive block was similar for both drugs. The nocifensive to motor block ratio was significantly higher with IQB-9302. CONCLUSIONS: IQB-9302 produced an anesthetic action similar to that of bupivacaine, although the former drug induced a slightly more potent nocifensive block. Nocifensive and motor block duration are very similar with IQB-9302, whereas bupivacaine induces a more prolonged motor block without nocifensive block. PMID- 11009241 TI - Effect of standard diluted epinephrine infusion on epidural anesthesia in labor. AB - BACKGROUND AND OBJECTIVES: Epinephrine is used with local anesthetics to prolong the duration of epidural analgesia and decrease the peak plasma concentrations of local anesthetics. However, the duration of labor may be prolonged because epinephrine reduces uterine activity. We designed a prospective, randomized, and doubleblind study to examine the effects of epinephrine infusion on the quality of analgesia and plasma concentration of local anesthetic, as well as the effect on the uteroplacental circulation, duration of the first or second stage of labor, and fetal outcome. METHODS: Twenty-four parturients received continuous epidural bupivacaine 0.125% (8 mL/h) combined either with epinephrine (40 microg/h) (n = 12) or without epinephrine (n = 12) for analgesia during labor. If patients requested additional analgesia, a bolus of 1% or 1.5% lidocaine (6 to 10 mL) was given. RESULTS: Only the plain bupivacaine group required additional lidocaine. However, epinephrine infusion prolonged the median (range) duration of the second stages of labor: 69 (21 to 231) minutes with epinephrine group versus 31 (8 to 99) minutes without epinephrine group (P < .05), and decreased pH in umbilical artery at the time of delivery. Epinephrine infusion did not change the uterine and umbilical blood flow, which were determined as the resistance indices. Changes in the fetal heart rate and Apgar score were also comparable. Epinephrine significantly reduced the umbilical venous to maternal arterial bupivacaine concentration (P < .05). CONCLUSIONS: A standard diluted epinephrine infusion (40 microg/h) into epidural space decreased anesthetic requirements. The possibility of the prolonged duration of labor remains a problem. PMID- 11009242 TI - Ondansetron for treatment of intrathecal morphine-induced pruritus after cesarean delivery. AB - BACKGROUND AND OBJECTIVES: Pruritus induced by intrathecal morphine is a concern in many obstetric patients after cesarean delivery and may detract from the benefit of postoperative pain relief. This study was performed to investigate the efficacy of ondansetron (5-HT3 receptor antagonist) in treatment of pruritus following intrathecal morphine. METHODS: Eighty parturients developing moderate to severe pruritus following intrathecal morphine were randomly allocated into 2 groups. One group received 4 mg ondansetron while the other group received placebo (normal saline). The improvement of pruritus and other adverse effects such as pain scores, nausea, vomiting, sedation, hallucination, and respiratory depression were determined at 30 minutes after study drugs' administration. RESULTS: The treatment success rate was higher in the ondansetron group than in the placebo group (80% v 36%, P < .001). Among the successfully treated patients, the recurrence rates of moderate to severe pruritus within 4 hours after administration of ondansetron and placebo were 12% and 70%, respectively (P < .001). The number of patients with decreased nausea and vomiting score was also higher in the ondansetron group (11 v 1, P < .006). CONCLUSION: Ondansetron treats intrathecal morphine-induced pruritus after cesarean delivery, particularly in patients suffering from both nausea/vomiting and pruritus. PMID- 11009243 TI - Lumbar epidurography. PMID- 11009244 TI - Neuropeptide-toxin conjugates in pain research and treatment. PMID- 11009245 TI - Maternal fever, neonatal sepsis evaluation, and epidural labor analgesia. AB - BACKGROUND AND OBJECTIVES: Numerous studies have found an association between epidural analgesia for labor and maternal fever (temperature > or =38 degrees C). Maternal fever often results in treatment with maternal or neonatal antibiotics, neonatal sepsis evaluation, and increased costs. METHODS: Medline was used to identify literature regarding the association between epidural labor analgesia and maternal fever/neonatal sepsis. Studies examining thermoregulation during pregnancy and/or epidural analgesia were also reviewed. RESULTS: There appears to be a strong association between epidural labor analgesia and maternal fever. The link between epidural labor analgesia and neonatal sepsis evaluation is less clear. The incidence of confirmed neonatal sepsis does not increase with maternal epidural analgesia. Causes of the association between epidural labor analgesia and maternal fever include selection bias, altered thermoregulation, and increased shivering or decreased sweating with epidural analgesia. CONCLUSIONS: Maternal epidural labor analgesia is associated with maternal fever and possibly increased neonatal sepsis evaluation. There is no proof the relationship is causal. PMID- 11009246 TI - Causal judgment from contingency information: the interpretation of factors common to all instances. AB - Participants made judgments about stimulus materials in which there were 2 possible causes of an outcome. One of these was a common factor, a factor present in all instances presented for judgment, and the other was a positive covariate of the outcome. Instead of interpreting the positive covariate as the cause, participants consistently preferred an interpretation in which the common factor was the cause and the positive covariate enabled the cause to produce its effect. Participants' judgments of both interpretations were predicted by the proportion of instances that were confirmatory for the interpretation and not by deltaP. The findings support a hypothesis that people have a multirole conceptualization of causality including, in addition to the roles of cause and effect, that of an enabler, a factor the presence of which ensures that a thing is in a state of readiness to produce a particular effect. PMID- 11009247 TI - Age of acquisition effects in adult lexical processing reflect loss of plasticity in maturing systems: insights from connectionist networks. AB - Early learned words are recognized and produced faster than later learned words. The authors showed that such age of acquisition effects are a natural property of connectionist models trained by back-propagation when patterns are introduced at different points into training and learning of early and late patterns is cumulative and interleaved. Analysis of hidden unit activations indicated that the age of acquisition effect reflects a gradual reduction in network plasticity and a consequent failure to differentiate late items as effectively as early ones. Further simulations examined the effects of vocabulary size, learning rate, sparseness of coding, use of a modified learning algorithm, loss of early items, acquisition of very late items, and lesioning the network. The relationship between age of acquisition and word frequency was explored, including analyses of how the relative influence of these factors is modulated by introducing weight decay. PMID- 11009248 TI - Task-set switching and long-term memory retrieval. AB - The authors tested the hypothesis of a close relationship between the intentional component of task-set switching ("advance reconfiguration;" R. D. Rogers & S. Monsell, 1995) and long-term memory (LTM) retrieval. Consistent with this hypothesis, switch costs are reported to be larger when the switched-to task involves high retrieval demands (i.e., retrieval of episodic information) than when it involves low retrieval demands (i.e., retrieval of semantic information). In contrast, switch costs were not affected by a primary-task difficulty manipulation unrelated to intentional retrieval demands (Experiment 2). Also, the retrieval-demand effect on switch costs was eliminated when time for advanced preparation or task cues explicitly specifying the task rules were provided (Experiment 3). Overall, results were consistent with the hypothesis that the intentional switch-cost component reflects the time demands of retrieving appropriate task rules from LTM. PMID- 11009249 TI - Similarity and inhibition in long-term memory: evidence for a two-factor theory. AB - Recalling a past experience often requires the suppression of related memories that compete with the retrieval target, causing memory impairment known as retrieval-induced forgetting. Two experiments examined how retrieval-induced forgetting varies with the similarity of the competitor and the target item (target-competitor similarity) and with the similarity between the competitors themselves (competitor-competitor similarity). According to the pattern suppression model (M. C. Anderson & B. A. Spellman, 1995), high target-competitor similarity should reduce impairment, whereas high competitor-competitor similarity should increase it. Both predictions were supported: Encoding target competitor similarities not only eliminated retrieval-induced forgetting but also reversed it, whereas encoding competitor-competitor similarities increased impairment. The differing effects of target-competitor and competitor-competitor similarity may resolve conflicting results concerning the effects of similarity on inhibition. PMID- 11009250 TI - The properties of retention intervals and their affect on retaining prospective memories. AB - Five experiments were conducted to explore how the character of the retention interval affected event-based prospective memory. According to the canons of retrospective memory, prospective performance should have been worse with increasing delays between intention formation and the time it was appropriate to complete an action. That result did not occur. Rather, prospective memory was better with increasing retention intervals in Experiments 1A, 1B, and 3. In manipulating the nature of the retention interval, the authors found that there were independent contributions of retention interval length and the number of intervening activities, with more activities leading to better prospective memory (Experiments 2 and 3). The identical retention intervals did not improve retrospective memory in Experiment 4. Theoretical explanations for these dissociations between prospective and retrospective memory are considered. PMID- 11009251 TI - Adult age differences in memory performance: tests of an associative deficit hypothesis. AB - An associative hypothesis to explain and predict older adults' deficient explicit episodic memory performance was outlined and tested. The hypothesis attributes a substantial part of older adults' deficient memory performance to their difficulty in merging unrelated attributes-units of an episode into a cohesive unit. Although each of the components can be memorized to a reasonable degree, the associations that tie the attributes-units to each other grow weaker in old age. Four experiments are reported that provide (a) a converging validity to the hypothesis by demonstrating this associative deficit for both interitem relationships and intraitem relationships and (b) a discriminant validity to the hypothesis by contrasting and testing competing predictions made by the associative hypothesis and by alternative hypotheses. The implications of these results to older adults' episodic memory performance are discussed. PMID- 11009252 TI - Interresponse times in serial recall: effects of intraserial repetition. AB - The authors examined the effects of intraserial repetition on multitrial serial learning of random consonant lists, analyzing both learning rates and perfect trial interresponse times (IRTs). Lists varied along 3 dimensions: list length, presence or absence of a repeated element, and lag between repeated elements. After achieving a forward-recall criterion on a given list, participants (N = 20) attempted backward recall. At small lags, IRTs between the repeated elements were very short (compared with IRTs from identical positions in nonrepetition lists). At larger lags, the IRT to recall the second repeated item was substantially longer than in control lists. These results reveal a latency analogue of the Ranschburg pattern seen in accuracy data. A Ranschburg pattern was also found in participants' learning rates. These results both generalize the Ranschburg phenomenon and present further challenges to theories of serial order memory. PMID- 11009253 TI - Repetition priming for familiar and unfamiliar faces in a sex-judgment task: evidence for a common route for the processing of sex and identity. AB - Repetition priming for faces was examined in a sex-judgment task given at test. Priming was found for edited, hair-removed photos of unfamiliar and familiar faces after a single presentation at study. Priming was also observed for the edited photos when study and test faces were different exemplars. Priming was not observed, however, when sex judgments were made at test to photos of complete, hair-included faces. These findings were interpreted by assuming that, for edited faces, internal features are attended, thereby activating face-recognition units that support performance. With complete faces, however, participants provided speeded judgments based primarily on the hairstyle. It is suggested that, for both familiar and unfamiliar faces, a common locus exists for the processing of the identity of a face and its sex. A single face-recognition model for the processing of familiar and unfamiliar faces is advocated. PMID- 11009254 TI - Gender stereotyping and decision processes: extending and reversing the gender bias in fame judgments. AB - M. R. Banaji and A. G. Greenwald (1995) demonstrated a gender bias in fame judgments--that is, an increase in judged fame due to prior processing that was larger for male than for female names. They suggested that participants shift criteria between judging men and women, using the more liberal criterion for judging men. This "criterion-shift" account appeared problematic for a number of reasons. In this article, 3 experiments are reported that were designed to evaluate the criterion-shift account of the gender bias in the false-fame effect against a distribution-shift account. The results were consistent with the criterion-shift account, and they helped to define more precisely the situations in which people may be ready to shift their response criterion on an item-by-item basis. In addition, the results were incompatible with an interpretation of the criterion shift as an artifact of the experimental situation in the experiments reported by M. R. Banaji and A. G. Greenwald. PMID- 11009255 TI - Surface form typicality and asymmetric transfer in episodic memory for spoken words. AB - A series of experiments was conducted to determine whether the typicality of the surface form of speech would affect memory retention of spoken words. For each surface characteristic studied, a continuous-recognition-memory task was used in which listeners based recognition judgments on word identity alone. For "typical" items, repetition benefits did not depend on whether the surface forms of the 1st and 2nd occurrences matched or mismatched. For "atypical" items, a larger repetition benefit occurred when the surface forms of the 2 occurrences matched. These results suggest that episodic memory for spoken words may be directly related to the perceived typicality of particular surface characteristics. PMID- 11009256 TI - The relative time course of semantic and phonological activation in reading Chinese. AB - The relative time course of semantic and phonological activation was investigated in the context of whether phonology mediates access to lexical representations in reading Chinese. Compound words (Experiment 1) and single-character words (Experiments 2 and 3) were preceded by semantic and phonological primes. Strong semantic priming effects were found at both short (57 ms) and long (200 ms) stimulus onset asynchrony (SOA), but phonological effects were either absent in lexical decision (Experiment 1), were present only at the longer SOA in character decision (Experiment 2) or were equally strong as semantic effects in naming (Experiment 3). Experiment 4 revealed facilitatory or inhibitory effects, depending on SOA, in phonological judgments to character pairs that were not phonologically but semantically related. It was concluded that, in reading Chinese, semantic information in the lexicon is activated at least as early and just as strongly as phonological information. PMID- 11009257 TI - Effects of sentence constraint on priming in natural language comprehension. AB - In 4 cross-modal naming experiments, researchers investigated the role of sentence constraint in natural language comprehension. On the sentence constraint account, incoming linguistic material activates semantic features that in turn pre-activate likely upcoming words. The 1st and 2nd experiments investigated whether stimulus offset asynchrony played a critical role in previous studies supporting the sentence constraint account. The 3rd and 4th experiments examined further predictions of the sentence constraint account, in particular whether pre activated words would compete for activation. In Experiment 3, the researchers manipulated whether an expected target word had a close competitor and found that response to the expected word was facilitated regardless of the proximity of a competitor. The 4th experiment established that close competitors were primed by the sentence frames and should have been available to compete with expected target words. Thus, word-level representations did not compete for activation. PMID- 11009258 TI - The cognate facilitation effect: implications for models of lexical access. AB - Do nonselected lexical nodes activate their phonological information? Catalan Spanish bilinguals were asked to name (a) pictures whose names are cognates in the 2 languages (words that are phonologically similar in the 2 languages) and (b) pictures whose names are noncognates in the 2 languages. If nonselected lexical nodes are phonologically encoded, naming latencies should be shorter for cognate words, and because the cognate status of words is only meaningful for bilingual speakers, this difference should disappear when testing monolingual speakers. The results of Experiment 1 fully supported these predictions. In Experiment 2, the difference between cognate and noncognate words was larger when naming in the nondominant language than when naming in the dominant language. The results of the 2 experiments are interpreted as providing support to cascaded activation models of lexical access. PMID- 11009259 TI - The role of mediators in memory retrieval as a function of practice: controlled mediation to direct access. AB - The role of prior knowledge in retrieval of Spanish-English vocabulary pairs learned using keyword mediators was examined in 4 experiments. Retrieval was tested immediately after learning and after 1-week and 1-month no-practice intervals (Experiment 1), after moderate retrieval practice (Experiment 2), and after extended retrieval practice (Experiments 3 and 4). Using accuracy, latency, and verbal report data, a detailed account of memory retrieval processes was developed. Initial retrieval is an explicit mediation process that involves retrieving keyword mediators into working memory and using them as retrieval cues to access the English equivalents of the Spanish words. After extended vocabulary retrieval practice, this sequential mediation process qualitatively changed to a direct retrieval process in which the English equivalent was accessed in a single working memory step. However, direct retrieval was still influenced by a covert mediation process. PMID- 11009260 TI - Counterfactual thinking about actions and failures to act. AB - When people think counterfactually about how a situation could have turned out differently, they mentally undo events in regular ways (e.g., they focus on actions not failures to act). Four experiments examine the recent discovery that the focus on actions in the short term switches to inactions in the long term. The experiments show that this temporal switch occurs only for particular sorts of situations. Experiment 1 showed no temporal pattern to the agency effect when 112 participants judged emotional impact and frequency of "if-only" thoughts from both short- and long-term perspectives for an investment scenario. Experiment 2 showed no temporal pattern when 190 participants considered a college choice scenario with a good outcome. Experiment 3 showed no temporal pattern when 131 participants considered an investment scenario even when the situation for the actor and nonactor was bad from the outset. Experiment 4, with 113 participants, showed a focus on actions even when the investment loss was equal for both the actor and nonactor. The implications of the results are discussed in terms of what is explicitly available in the mental representation of actions and inactions. PMID- 11009261 TI - Assessing the empirical validity of the "take-the-best" heuristic as a model of human probabilistic inference. AB - The boundedly rational 'Take-The-Best" heuristic (TTB) was proposed by G. Gigerenzer, U. Hoffrage, and H. Kleinbolting (1991) as a model of fast and frugal probabilistic inferences. Although the simple lexicographic rule proved to be successful in computer simulations, direct empirical demonstrations of its adequacy as a psychological model are lacking because of several methodical problems. In 4 experiments with a total of 210 participants, this question was addressed. Whereas Experiment 1 showed that TTB is not valid as a universal hypothesis about probabilistic inferences, up to 28% of participants in Experiment 2 and 53% of participants in Experiment 3 were classified as TTB users. Experiment 4 revealed that investment costs for information seem to be a relevant factor leading participants to switch to a noncompensatory TTB strategy. The observed individual differences in strategy use imply the recommendation of an idiographic approach to decision-making research. PMID- 11009262 TI - Impact of end-stage renal disease on clinical and angiographic outcomes after coronary stenting. AB - Although patients with end-stage renal disease (ESRD) are at high risk for restenosis that requires repeat revascularization after balloon angioplasty, their restenosis rate after coronary stenting is still unknown. Over a 4-year period, we performed coronary stenting on 40 lesions in 34 patients with ESRD. We compared these lesions with 80 lesions from patients without renal disease who underwent coronary stenting in the STARS and WINS clinical trials, matched for treatment site, diabetes, lesion length, and reference vessel diameter. Quantitative coronary angiography was performed on all lesions and clinical outcomes were assessed at 9-month follow-up. Clinical and angiographic characteristics were well matched between the 2 groups and acute clinical success rates were similar. Despite comparable initial angiographic results over the 9 month follow-up period, repeat target lesion revascularization was twice as frequent in the ESRD group compared with the control group (35% vs 16%, p <0.05). After adjusting for differences in postprocedural minimum lumen diameter and other angiographic and clinical characteristics, ESRD remained the most important predictor of late target lesion revascularization (relative risk = 2.3, p = 0.04). In addition, overall 9-month mortality was higher for ESRD patients than for the control population (18% vs 2%, p <0.01). Thus, despite similar angiographic results, patients with ESRD are at higher risk for target lesion revascularization after coronary stenting than controls. Nonetheless, most patients with ESRD do not develop restenosis after stent placement, suggesting an important role for stenting in the management of this challenging population. PMID- 11009263 TI - Effect of coronary artery bypass surgery on myocardial perfusion and ejection fraction response to inotropic stimulation in patients without improvement in resting ejection fraction. AB - The aim of this study was to assess the effect of coronary artery bypass grafting (CABG) on myocardial perfusion and left ventricular (LV) contractile reserve in patients with reduced ejection fraction (EF). We studied 57 patients (age 59 +/- 8 years, 46 men and 11 women) with EF < or = 40% referred for CABG with dobutamine (up to 40 microg/kg/min) stress-reinjection thallium-201 single-photon emission computed tomography, and radionuclide ventriculography at rest and at low-dose dobutamine before and 3 months after CABG. An increase in resting EF > or = 5% occurred in 12 patients (group A) after CABG (EF 34% before and 46% after CABG), whereas no increase occurred in the remaining 45 patients (group B) (EF 34% before and 32% after CABG). A significant increase in EF from rest to low dose dobutamine radionuclide ventriculography occurred before and after CABG. The magnitude of increase was more significant after than before CABG in group A (12% vs 7%) as well as in group B (13% vs 7%, both p <0.001). Patients in both groups had a significant reduction in stress, rest, and ischemic perfusion scores after CABG. However, the percentage of reduction in resting perfusion defect score was more significant in group A than in group B (60% vs 30%, respectively, p <0.01). It is concluded that CABG induces a significant improvement in resting myocardial perfusion and EF response to inotropic stimulation, even in the absence of improved EF at rest. Patients without improvement in resting EF after CABG have mild improvement in resting myocardial perfusion that may be sufficient to increase EF after CABG during inotropic stimulation, but not at rest. We describe the myocardium with these characteristics as "the reactive myocardium." PMID- 11009264 TI - Coronary artery calcium evaluation by electron beam computed tomography and its relation to new cardiovascular events. AB - Electron beam computed tomography is widely used to screen for coronary artery calcium (CAC). We evaluated the relation of CAC to future cardiovascular disease events in 926 asymptomatic persons (735 men and 191 women, mean age 54 years) who underwent a baseline electron beam computed tomographic scan. All subjects included in this report returned a follow-up questionnaire 2 to 4 years (mean 3.3) after scanning, inquiring about myocardial infarction, stroke, and revascularizations. Sixty percent of men and 40% of women had a positive scan at baseline. Twenty-eight cardiovascular events occurred and were confirmed by blinded medical record review. The presence of CAC (a total calcium score of >0) and increasing score quartiles were related to the occurrence of new myocardial infarction (p <0.05), revascularizations (p <0.001), and total cardiovascular events (p <0.001). Those with scores at or above the median (score of 5) had a relative risk of 4.5 (p <0.01) for new events. From Cox regression models, adjusted for age, gender, and coronary risk factors, the relative risks for those with scores of 81 to 270 and -271 (compared with 0) for cardiovascular events were 4.5 (p <0.05) and 8.8 (p <0.001), respectively. These data support previous reports showing CAC to be a modest predictor of future cardiovascular events. PMID- 11009265 TI - Effects of cilostazol on angiographic restenosis after coronary stent placement. AB - This study evaluates the impact of cilostazol on post-stenting restenosis. Cilostazol is a potent antiplatelet agent with antiproliferative properties. Few data are available about the effect of cilostazol on poststenting restenosis. Four hundred nine patients (494 lesions) who were scheduled for elective stenting were randomized to receive aspirin plus ticlopidine (group I, n = 201, 240 lesions) or aspirin plus cilostazol (group II, n = 208, 254 lesions), starting 2 days before stenting. Ticlopidine was given for 1 month and cilostazol for 6 months. Follow-up angiography was performed at 6 months, and clinical evaluation at regular intervals. Baseline characteristics were similar between the 2 groups. The procedural success rate was 99.6% in group I and 100% in group II. There were no cases of stent thrombosis after stenting. Angiographic follow-up was performed in 380 of the 494 eligible lesions and the angiographic restenosis rate was 27% in group I and 22.9% in group II (p = NS). However, diffuse type in-stent restenosis was more common in group I than in group II (54.2% vs 26.8%, respectively, p <0.05). In diabetic patients, the angiographic restenosis rate was 50% in group I and 21.7% in group II (p <0.05). Clinical events during follow up did not differ between the 2 groups. In conclusion, aspirin plus cilostazol seems to be an effective antithrombotic regimen with comparable results to aspirin plus ticlopidine, but it does not reduce the overall angiographic restenosis rate after elective coronary stenting. PMID- 11009266 TI - Cerebral hemodynamics in carotid sinus syndrome and atrioventricular block. AB - Carotid sinus syndrome (CSS) is a cause of syncope due to exaggerated baroreceptor-mediated cardioinhibitory/vasodepressive reflexes. We sought to determine if cerebral hemodynamics and regulation were specifically altered in these patients by comparison with pure asystole without vasodepression in patients with atrioventricular block (AVB). Mean blood flow velocity (transcranial Doppler sonography) and mean arterial blood pressure (Finapres) were recorded during cardioinhibition induced by carotid massage in patients with CSS (n = 14, 75 +/- SD 8 years) and asystole induced by temporary pacemaker inhibition in patients with complete AVB (n = 10, 69 +/- 11 years). Cerebrovascular resistance was estimated by the arterial pressure/cerebral flow velocity ratio, and dynamic cerebral autoregulatory responses were determined by the rate of regulation and autoregulatory index. Asystole and cardioinhibition each induced a decrease in arterial pressure (CSS 55 +/- 9% vs AVB 40 +/- 14%, p <0.05) and cerebral flow velocity (CSS 66 +/- 19% vs AVB 69 +/- 14%, p = NS), with an initial transient increase in cerebrovascular resistance (CSS 102 +/- 136% vs AVB 128 +/- 92%, p = NS) followed by a decrease (CSS 38 +/- 12%, AVB 29 +/- 13%, p = NS). The rate of regulation and autoregulatory index were higher with AVB (0.43 +/- 0.20 and 8.5 +/- 1.1 second(-1)) than CSS (0.20 +/- 0.12 and 4.8 +/- 1.3 second(-1), respectively, p <0.01 and p <0.001 vs AVB). During asystole and vasodepression, cerebral hypoperfusion in CSS is normally compensated for by cerebral autoregulation. The lower rate of regulation in CSS compared with AVB likely results from persistent peripheral vasodepression triggered by carotid massage. PMID- 11009267 TI - Subclinical left ventricular dysfunction in systemic hypertension and the role of 24-hour blood pressure. AB - The relation between blood pressure (BP) and left ventricular (LV) systolic function in systemic hypertension is controversial. We assessed the relation of LV midwall fractional shortening (FS) to 24-hour BP in 1,702 never-treated hypertensive subjects (age 48 +/- 12 years), who underwent 24-hour BP monitoring and echocardiography. Stress-corrected endocardial and midwall FS (the latter calculated taking into account the epicardial migration of midwall during systole) were predicted in hypertensives on the basis of the values observed in 130 healthy normotensives (age 43 +/- 13 years, office BP 126/78 mm Hg). Subjects below the fifth percentile of observed-to-predicted FS had depressed LV function. The use of midwall FS resulted in an increase from 3.5% to 17.5% in the proportion of patients with depressed chamber function. Compared with the group with normal function, subjects with low midwall LV function had similar office systolic BP (155 +/- 21 vs 154 +/- 17 mm Hg), but increased 24-hour systolic BP (140 +/- 17 vs 133 +/- 12 mm Hg, p <0.001). Midwall FS had a closer negative relation to 24-hour systolic BP than to office systolic BP (r = -0.27 vs -0.08, p <0.001), whereas this difference was not apparent for diastolic BP (r = -0.23 vs 0.20). Compared with endocardial FS, midwall FS had a stronger inverse association to LV mass (r = -0.45 vs -0.16, p <0.001). Thus, an increased 24-hour BP load may chronically lead to depressed myocardial function in systemic hypertension in the absence of clinically overt heart disease. PMID- 11009268 TI - Effects of pravastatin on left ventricular mass in patients with hyperlipidemia and essential hypertension. AB - Left ventricular (LV) mass is a powerful predictor for future cardiovascular events. Epidemiologic studies have shown that hyperlipidemia is associated with higher LV mass. The effects of statin therapy for hyperlipidemia on LV mass have not been studied. To determine the effects of statin therapy on LV mass, we prospectively studied 3 groups of age and body surface area-matched patients: group 1 (n = 20), patients with systemic hypertension and hyperlipidemia treated with pravastatin plus anti-hypertensive drugs; group 2 (n = 20), patients with hypertension and hyperlipidemia treated with hypertensive agents and diet control alone; and group 3 (n = 20), hypertensive patients with normolipidemia treated with antihypertensive agents. A group of controls without hypertension or hyperlipidemia was used for comparison. Echocardiograms were recorded at baseline and after 6-month therapy. All hypertensive groups showed significant decreases in LV mass index after treatment. Group 1 had the greatest decrease in LV mass and it was significantly higher than in groups 2 and 3. Multivariate analysis revealed that regression of LV mass was significantly correlated only with the use of statins and sex (p = 0.005 and 0.01, respectively, R(2) = 0.47). Linear regression analysis in group 1 showed a significant correlation between changes in arterial compliance and LV mass regression (r = 0.57, p = 0.01). Thus, the addition of a statin may have an additional effect on reducing LV mass, independent of lipid-lowering effects. PMID- 11009269 TI - Containment of heart failure hospitalizations and cost by angiotensin-converting enzyme inhibitor dosage optimization. AB - Using our model relating angiotensin-converting enzyme (ACE) inhibitor dosing and outcomes in heart failure (HF), we designed a prospective intervention trial for patients with systolic dysfunction. A clinical pharmacist initiated or titrated ACE inhibitor therapy or adjusted other medications within an HF management program based on Agency for Healthcare Policy and Research guidelines. Entry into the protocol required the approval of the attending physician. All patients received dietary, nursing, rehabilitation, social service, and clinical pharmacy consultations. Treatment conformed to Agency for Healthcare Policy and Research guidelines in 25% of patients (group A). Suboptimal therapy (75% of patients) was usually due to failure to administer an ACE inhibitor (48%) or inadequate dosing of an ACE inhibitor (46%). In 62% of suboptimal cases, the attending physician agreed to follow the clinical pharmacist's recommendations (group B). Patients of physicians who declined pharmacist intervention served as a negative control (group C). On admission, mean enalapril-equivalent daily doses in groups A, B, and C were 30, 4, and 6 mg, respectively, and at discharge, 36, 18, and 6 mg, respectively. At 180 days, rehospitalization frequency and total charges were lower in groups A (31% and $5,600) and B (35% and $3,800) than in group C (63% [p <0.004] and $9,800 [p <0.04]). Thus, optimization of ACE inhibitor doses by a clinical pharmacist can greatly improve rehospitalization rates and significantly lower cost of care in an HF management program. PMID- 11009270 TI - Transcardiac extraction of circulating endothelin-1 across the failing heart. AB - To determine the transcardiac gradient of plasma endothelin-1 (ET-1) in patients with congestive heart failure (CHF), we measured plasma levels of ET-1 in both the aortic root and the coronary sinus in 14 normal subjects and 79 consecutive patients with CHF. In normal subjects, plasma ET-1 was significantly higher in the coronary sinus than in the aortic root; these findings were also shown in patients with mild CHF, suggesting that there was ET-1 spillover across the heart. In contrast, plasma ET-1 was significantly lower in the coronary sinus than in the aortic root in patients with severe CHF, suggesting there was ET-1 extraction across the heart in patients with severe CHF. The transcardiac gradient of plasma ET-1 was correlated with the left ventricular end-diastolic volume index (r = 0.501, p <0.0001) and plasma level of procollagen type III amino terminal peptide in the coronary sinus (r = 0.54, p = 0.0008), a marker of myocardial fibrosis. Stepwise multivariate analysis showed that the transcardiac gradient of plasma ET-1 was an independent and significant relation with the left ventricular end-diastolic volume index in patients with CHF (r = 0.665, p <0.0001). These findings suggest that elevated circulating ET-1 is extracted across the failing heart with a significant correlation between the transcardiac gradient of plasma ET-1 and the left ventricular end-diastolic volume index, suggesting that ET receptors are upregulated in the failing ventricle and that the elevated circulating ET-1 might stimulate the process of left ventricular remodeling in patients with severe CHF. PMID- 11009271 TI - Predictors of left atrial spontaneous echo contrast and thrombi in patients with mitral stenosis and atrial fibrillation. AB - The purpose of this study was to investigate the independent factors associated with the presence of left atrial (LA) spontaneous echo contrast (SEC) and thromboembolic events in patients with mitral stenosis (MS) in chronic atrial fibrillation (AF). Factors independently associated with LASEC, thrombi, and embolic events have been mainly investigated in patients with nonvalvular AF or inhomogeneous populations with rheumatic heart disease. Transesophageal and transthoracic echo studies were performed in 129 patients with MS in chronic AF. Previous embolic events were documented in 45 patients, 20 of them within 6 months, and 65 patients were receiving long-term anticoagulation. The intensity of LASEC and mitral regurgitation, the presence of thrombi and active LA appendage flow (peak velocities > or = 20 cm/s), and LA volume as well as other conventional echo-Doppler determinations were investigated in every patient. The prevalences of significant LASEC (degrees 3+ and 4+), thrombus, active LA appendage flow, and significant mitral regurgitation (>2+) were: 52% (67 patients), 29.5% (38 patients), 32% (41 patients), and 36% (47 patients), respectively. Multivariate analysis showed that decreasing mitral regurgitation severity, absence of active LA appendage flow, and mitral valve area were the independent correlates of LASEC (odds ratio [OR] 3.7, 5.4, and 0.17, respectively; all p <0.02). Active LA appendage flow and anticoagulant therapy were associated negatively, whereas the severity of LASEC was associated positively with the finding of LA thrombus (OR 9.6, 3.9, and 1.6, respectively; all p <0.05). The intensity of LASEC and previous anticoagulant therapy (OR 1.74 and 4.5, respectively; p <0.005) were the independent covariates of thrombi and/or recent embolic events. In conclusion, the severity of mitral regurgitation and lack of active LA appendage flow were, respectively, the strongest independent correlates of significant LASEC and thrombus in patients with MS in chronic AF. LASEC remains the cardiac factor most strongly associated with thrombus and/or recent embolic events in these patients. PMID- 11009272 TI - Prevalence and diagnostic value of precordial murmurs for valvular regurgitation in obese patients treated with dexfenfluramine. AB - Echocardiography is recommended for the detection of valvular regurgitation in asymptomatic users of anorexigens with a heart murmur. To determine the prevalence and diagnostic value of heart murmurs for valvular regurgitation, 223 patients receiving dexfenfluramine therapy for 6.9 months and 189 matched controls underwent history and cardiac auscultation by experienced noncardiologists unaware of echocardiography. Color Doppler echocardiograms were interpreted by 3 observers unaware of patients' clinical data. The frequency of at least mild regurgitation of any valve and abnormal regurgitation (moderate mitral or tricuspid or mild aortic regurgitation) were determined. Heart murmurs heard in 31 dexfenfluramine users (14%) and in 20 controls (11%) were all systolic and of grade I to II/VI intensity. Mild or worse regurgitation of any valve showed a trend in patients (18% vs 11.6% in controls, odds ratio [OR] 1.66, confidence interval [CI] 0.95 to 2.9, p = 0.08), but abnormal regurgitation (includes Food and Drug Administration grade regurgitation) was more common in patients (9% vs 3% in controls, OR 3.0, CI 1.18 to 7.65, p = 0.02). In dexfenfluramine users, heart murmurs were associated with at least mild or abnormal regurgitation (OR 3.1 and 3.05, 95% CI 1.34 to 7.13 and 1.1 to 8.67; p = 0.008 and 0.036, respectively), had a specificity of 89% and 88%, negative predictive value of 85% and 93%, but sensitivity of 37% and 30%, and positive predictive value of 35% and 19%, respectively. Most valves missed by cardiac auscultation had normal morphology and mild regurgitation. Finally, heart murmurs had better diagnostic value for either type of valvular regurgitation than heart murmurs and clinical variables or clinical variables alone. In summary, in dexfenfluramine users the prevalence of heart murmurs was low and their absence predicted absence of mild or worse regurgitation of any valve or abnormal valvular regurgitation. Therefore, cardiac auscultation should be the screening method of choice for detecting valvular regurgitation in users of anorexigens. PMID- 11009274 TI - Conclusions from the VA-HIT study. PMID- 11009273 TI - Evolution of aortic regurgitation following simple patch closure of doubly committed subarterial ventricular septal defect. AB - We reviewed the Doppler echocardiographic findings of the aortic valve and associated aortic regurgitation (AR) in 55 patients who underwent patch closure of doubly committed subarterial ventricular septal defect (VSD). The maximal diameter of the VSD measured > or = 5 mm, whereas the postoperative follow-up interval was > or = 5 years. Twenty-three patients underwent closure before they developed aortic cusp prolapse or AR (group A). In 15 patients the VSD was closed when aortic cusp prolapse was recognized, but AR was absent (group B). Aortic cusp prolapse with AR was detected before closure in a further 15 patients (group C). Of 8 patients with no AR before closure, AR was detected during follow-up in 6 group A and in 2 group B patients. In group C, AR resolved after surgery in 4 patients, whereas AR grade improved in a further 8 patients and remained unchanged in 3. Although residual AR was more frequent in patients with aortic cusp prolapse and AR before closure, it was silent and asymptomatic. PMID- 11009275 TI - Should the split format in the third position of the pacemaker code be resurrected? AB - It is possible to characterize some of the sensing functions of new multisite pacing systems by resurrecting the split format in the third position of the standard pacemaker code. This approach permits accurate representation of the horizontal and vertical triggering functions of multisite dual-chamber pacemakers without creating a new code. PMID- 11009276 TI - High-density lipoprotein cholesterol and triglyceride response with simvastatin versus atorvastatin in familial hypercholesterolemia. AB - The clinical and biochemical determinants of high-density lipoprotein (HDL) and triglyceride response to simvastatin and atorvastatin were assessed in 150 patients with severe hyperlipidemia treated in a randomized open-trial format design. Triglyceride reduction was only dependent on HDL:apolipoprotein A1, change in apolipoprotein B, and dose response, whereas an increase in HDL was dependent on initial LDL, change in LDL or dose response, and therapy with simvastatin. PMID- 11009277 TI - Effect of probucol on repeat revascularization rate after percutaneous transluminal coronary angioplasty (from the Probucol Angioplasty Restenosis Trial [PART]). AB - To address the issue of whether probucol reduces clinical events after percutaneous transluminal coronary angioplasty (PTCA), we surveyed clinical status at 1 year after PTCA of 101 patients who had entered the Probucol Restenosis Angioplasty Trial. Repeat angioplasty at index lesions were required in 5 patients in the probucol group and in 12 in the control group, suggesting that probucol administered beginning 4 weeks before PTCA reduces repeat revascularization rates for 1 year. PMID- 11009278 TI - Impact of enoxaparin low molecular weight heparin in patients with Q-wave myocardial infarction. AB - A subgroup meta-analysis from the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-Wave Coronary Events (ESSENCE) and the Thrombolysis in Myocardial Infarction (TIMI) 11B studies has shown that enoxaparin is superior to unfractionated heparin in reducing the composite end points of death, myocardial infarction, and emergency revascularization in patients with Q-wave myocardial infarction. The beneficial treatment effect was significant at 43 days. PMID- 11009279 TI - Association between heart rate-corrected QT interval and coronary risk factors in 2,894 healthy subjects (the DESIR Study). Data from an Epidemiological Study on the Insulin Resistance syndrome. AB - In the prospective Data from an Epidemiological Study on the Insulin Resistance Syndrome, 2,894 healthy subjects aged 30 to 64 years had determinations of fasting glucose, insulin, serum lipid and fibrinogen concentrations, blood pressures, body mass index, and waist-hip ratio, as well as tobacco and alcohol consumptions and physical activity. A 12-lead electrocardiogram with automatic measurement of the QT interval was recorded and the formula used for heart rate correction was based on the best-fit regression between QT and heart rate. The QT duration was influenced by glucose homeostasis in both sexes, and increased in men with physical activity; there was a dose-effect relation for men who smoked. PMID- 11009280 TI - Continuous intravenous diltiazem infusion for short-term ventricular rate control in children. AB - Intravenous diltiazem was administered to 10 pediatric patients with primary atrial tachyarrhythmias with rapid ventricular response. Rapid, consistent, and safe temporary ventricular rate control was obtained in all patients given this medication. PMID- 11009281 TI - Holter monitoring: are two days better than one? AB - The optimal duration of Holter monitoring (HM) to minimize costs and maximize yield is unknown. In a retrospective review of 164 patients referred to a tertiary care center for evaluation with 2 days of HM, we found that 48 hours was not cost effective when compared with the traditional 24-hour period. PMID- 11009282 TI - Effect of hypertension on cardiac mass and radial artery wall thickness. AB - In subjects with essential hypertension, increased cardiac and radial artery mass are positively associated. The mechanical factor that contributes in the larger extent to their enhancement is pulse pressure, not mean arterial pressure. PMID- 11009283 TI - Usefulness of left atrial size in predicting postoperative symptomatic improvement in patients with aortic stenosis. AB - Although surgery is highly effective for symptomatic relief in patients with aortic stenosis, symptoms of congestive heart failure may be still present postoperatively. This group of patients with aortic stenosis is characterized by a wide range of left atrial size, which can predict postoperative symptomatic improvement. PMID- 11009284 TI - Effects of milrinone on pulmonary gas exchange in catecholamine-dependent heart failure. AB - Hemodynamic benefits of milrinone administration are accompanied by adverse effects on arterial oxygenation in mechanically ventilated patients with end stage heart failure. Particular attention should be focused on pulmonary gas exchange variables after initiation of milrinone treatment in the critically ill patient. PMID- 11009285 TI - Impact of tissue harmonic imaging on the assessment of rheumatic mitral stenosis. AB - We examined the effect of tissue harmonic imaging on the echocardiographic splitability score and valve area measurement of 40 patients with rheumatic mitral stenosis. Planimetered valve areas were unaffected by the use of harmonic imaging, but valve scores were increased, particularly in patients with scores <10. PMID- 11009286 TI - Mechanism of pulmonary vein stenosis in infants with normally connected veins. AB - We used microscopy, immunohistochemistry, and cell culture to identify the mechanism of restenosis in 4 infants with isolated pulmonary vein stenosis. Recurrent obstruction appears to be due to myofibroblastic proliferation in this fatal disease. PMID- 11009287 TI - Six-month success of intracoronary stenting for anomalous coronary arteries associated with myocardial ischemia. AB - Fourteen patients with anomalous coronary arteries associated with objective evidence of myocardial ischemia, a group at increased risk for cardiac events, were successfully stented. This led to 6-month patency and resolution of the ischemia, which was confirmed angiographically and by stress imaging studies. PMID- 11009289 TI - Increased plasma homocysteine is an independent predictor of new atherothrombotic brain infarction in older persons. AB - A prospective study investigated the association of plasma homocysteine and other risk factors with the incidence of atherothrombotic brain infarction (ABI) at 31 +/- 9 month follow-up in 153 men and 347 women (mean age 81 +/- 9 years, median age 82). The stepwise Cox regression model showed that significant independent predictors of new ABI in older persons were age (risk ratio 1.060 for each 1-year increase of age), plasma homocysteine (risk ratio 1.079 for each 1 micromol/L increase), prior ABI infarction (risk ratio 3.282), current cigarette smoking (risk ratio 2.687), hypertension (risk ratio 2.965), and diabetes mellitus (risk ratio 2.015). PMID- 11009288 TI - Comparison of quinapril versus atenolol: effects on blood pressure and cardiac mass after renal transplantation. AB - Based on epidemiologic facts on elevated cardiovascular mortality in renal allograft recipients, an echocardiographic 2-year follow-up in hypertensive renal allograft recipients was conducted. This study provides evidence that, in contrast to atenolol, quinapril, independent of blood pressure reduction, reduces left ventricular hypertrophy and improves left ventricular diastolic function in this population. PMID- 11009290 TI - Effect of shaving the chest of hirsute subjects on transthoracic impedance to self-adhesive defibrillation electrode pads. AB - We determined transthoracic impedance (TTI) before and after shaving in 9 hirsute and 11 nonhirsute subjects. TTI in hirsute subjects was mean +/- SE 162 +/- 11 versus 103 +/- 6 ohms in nonhirsute subjects (p <0.01); TTI decreased to 105 +/- 3 ohms after shaving the hirsute chests (p <0.01). PMID- 11009291 TI - Postmenopausal smokers show reduced hemodynamic benefit from oral hormone replacement. AB - With use of a randomized, placebo-controlled trial, 62 healthy, postmenopausal smokers and nonsmokers were tested for resting and stress-induced hemodynamic variables before and after 6 months of treatment with either oral hormone replacement therapy or placebo. Smokers had significantly less reduction in both resting and stress-induced vascular resistance and blood pressure after treatment with oral hormone replacement therapy than nonsmokers. PMID- 11009292 TI - Statins, high-density lipoprotein, and the low-density lipoprotein/high-density lipoprotein ratio. PMID- 11009293 TI - Pediatric radiologists: who we are and what we do. Results of a membership survey of the Society for Pediatric Radiology--1999. AB - BACKGROUND: There is a need for reliable monitoring of workforce trends in the field of pediatric radiology by the Society for Pediatric Radiology. In addition, the Society should periodically assess itself as to its mission and relevance to its members via membership surveys. OBJECTIVE: The Membership Committee of the Society for Pediatric Radiology, 1999, conducted a 54-question survey to determine the makeup of its members, job profiles, satisfaction with services of the Society for Pediatric Radiology, and its official journal, Pediatric Radiology. MATERIALS AND METHODS: Seven hundred fifty surveys were given to active members of the Society for Pediatric Radiology in the United States and Canada. There were 275 surveys returned for an overall response of 37%. RESULTS: Mean age of members is 48 years with 99% of respondents working. Membership is 34% female and 66% male. Women members of the Society are younger (45 vs 49 years, P = 0.0012) and work less hours (47.8 vs 51.0, P = 0.0135) than men. Fifty seven percent of respondents practice in a freestanding children's hospital, 29% in a "children's hospital within a hospital," and 14% are in community hospitals or an office-based practice. Eighty-two percent of the responding pediatric radiologist's time is spent in performing examination on children, with only 18% spent on adult work or administration. Forty-eight percent work at more than one office. Sixty-one percent worked evenings or weekends, excluding night call. Two hundred twenty-two of 275 respondents had received a Certificate of Added Qualification. The meeting "for CME credit" was considered the most important benefit of Society membership. Most respondents read select articles in Pediatric Radiology. There was sentiment to decrease esoteric case reports in favor of review articles. Only 19% of respondents submitted their articles to Pediatric Radiology initially. CONCLUSION: Pediatric radiologists are a diverse membership with the common goal of advocating for healthcare and imaging in children. Survey information given to the leadership of the Society will help the organization remain responsive to its members. PMID- 11009294 TI - What is the normal prevalence of vesicoureteral reflux? AB - BACKGROUND: The prevalence of vesicoureteral reflux in normal children has been estimated to be 0.4% to 1.8%. Based on epidemiological data, it can be estimated that 2.2% of girls and 0.6% of boys may be found to have reflux as a result of the investigation of urinary tract infection. This implies that most children with reflux will develop urinary tract infection. However, most children with reflux detected by screening do not develop infection. Hence the prevalence of vesicoureteral reflux in normal children may be higher than the previous estimates. OBJECTIVE: To determine the prevalence of vesicoureteral reflux in groups of children with different clinical indications for cystogram. MATERIALS AND METHODS: Over 250 articles from the literature were reviewed to determine the prevalence of vesicoureteral reflux in children undergoing cystogram. Means and 95% confidence limits were derived from the data in studies that met the inclusion criteria. RESULTS: The prevalence of vesicoureteral reflux in children with urinary tract infection was 31.1% (95% CI: 29.9-32.8). The prevalence of vesicoureteral reflux varied between patient groups, but often approached or exceeded the prevalence in urinary tract infection. The prevalence in normal kidneys was 17.2% (95% CI: 14.4-20.1). CONCLUSION: Vesicoureteral reflux is common in nearly all patient groups examined. The prevalence of vesicoureteral reflux in normal children is probably significantly higher than the traditional estimates. PMID- 11009295 TI - The impact of imaging in the management of intussusception owing to pathologic lead points in children. A review of 43 cases. AB - OBJECTIVE: To review the imaging appearances, management and outcome of a large number of children with intussusception owing to pathologic lead points (PLP) in an attempt to define the role of various imaging modalities in this clinical setting. MATERIALS AND METHODS: Review of the records and imaging studies of 43 children with intussusception due to PLP diagnosed between 1986 and 1999. RESULTS: The commonest PLP found were Meckel diverticulum, polyps, Henoch Schonlein purpura and cystic fibrosis. PLP were depicted on sonography in 23 (66%) of 35 patients, on computed tomography in 5 (71%) of 7, on air enema in 3 (11%) of 28, and on barium enema in 6 (40%) of 15. Air enema successfully reduced 60% of the intussusceptions. Nine children had recurrent intussusceptions. CONCLUSION: Sonography depicted two-thirds of PLP and provided a specific diagnosis in nearly one-third of our series. Our review does not provide sufficient data on how to continue the investigation of those patients in whom sonography does not depict a PLP but in whom there is a high index of suspicion for its presence. It remains a diagnostic challenge as to how to search for PLP in these patients, and other imaging modalities have to be requested according to each particular case. PMID- 11009296 TI - Aneurysmal dilatation of the paraumbilical vein in an infant. AB - The paraumbilical veins are one of the most common collateral pathways developing in portal hypertension. Aneurysmal dilatation of a patent paraumbilical vein is a very unusual finding. We report the first infant case of a paraumbilical vein aneurysm and its Doppler ultrasound findings. PMID- 11009297 TI - Transvenous coil embolization of an extrahepatic portosystemic shunt in a dog: a naturally occurring model of portosystemic malformations in humans. AB - Congenital patent ductus venosus (PDV) occurs far more commonly in dogs than in people; consequently, the natural course of the disease in dogs was studied as a model to understand the pathophysiology behind the vascular anomaly and its response to therapy better. In this report, the authors describe the results of percutaneous coil embolization as a single procedure in a dog with a single congenital extrahepatic portocaval shunt and compare portosystemic vascular anomalies (PSVA) seen in dogs with those seen in children. PMID- 11009298 TI - Pancreatic pseudotumor in an 11-year-old child: imaging findings. AB - BACKGROUND: An inflammatory pseudotumor is a benign, solid lesion of unclear etiology. Some authors believe it is a true neoplasm, while others consider it a post-infectious or post-traumatic process. It is most commonly found in the lung; an inflammatory pseudotumor of the pancreas is rare. This case report is the sixth of a pancreatic pseudotumor in a child. PATIENTS AND METHODS: An 11-year old girl presented with obstructive jaundice due to a mass in the head of the pancreas. The mass was identified by sonography. This was confirmed by MRI and CT. The mass enhanced with gadolinium, but its enhancement at CT was similar to the remainder of the pancreas. RESULTS: At operation, a pancreatic inflammatory pseudotumor was totally resected. PMID- 11009299 TI - Further report of small-bowel intussusceptions related to gastrojejunostomy tubes. AB - BACKGROUND: The use of gastrojejunostomy (GJ) tubes for feeding pediatric patients who have inadequate oral intake, cannot tolerate gastric feeding, or have significant gastroesophageal reflux may present problems. OBJECTIVE: To report our experience of intussusception associated with GJ tubes, with emphasis on risk factors. MATERIALS AND METHODS: Clinical histories and imaging studies were reviewed for all patients with GJ tube-associated intussusceptions at our institution from January 1995 to March 1999. RESULTS: Of 251 GJ tubes inserted, 40 (16%) intussusceptions occurred in 30 patients (20 males). They ranged in age from 3 months to 17 years (mean 2.6 years) and in weight from 3 to 90 kg (mean 12.5 kg). The underlying diagnosis varied. The main symptom was bilious vomiting. The initial diagnosis was made by sonography in 19 cases and fluoroscopy in 21. Intussusceptions recurred eight times in patients with a distal pigtail (n = 17) but only twice in patients without the distal pigtail (n = 18). CONCLUSIONS: GJ tube related intussusception is a common, easily diagnosed problem. Predisposing factors appear to be male sex, young children, and presence of a distal pigtail on the tube. Awareness of intussusception is imperative for prompt diagnosis to achieve feeding tolerance. PMID- 11009300 TI - Intracranial resistive index (RI) values in normal term infants during the first day of life. AB - PURPOSE: The purpose of this study was to determine normal resistive index (RI) values for term neonates during the first day of life as part of an ongoing prospective study of RI values in term infants with perinatal asphyxia. MATERIALS AND METHODS: Forty normal term neonates underwent cranial sonography and Doppler during the first 24 h after birth. Transfontanelle Doppler was performed of the internal carotid, anterior cerebral, and middle cerebral arteries bilaterally. In addition, transtemporal Doppler was performed of the middle cerebral arteries bilaterally. Mean and median RI values were calculated in all vessels interrogated. The transfontanelle and transtemporal middle cerebral artery measurements were compared using paired t-tests. RESULTS: The overall mean RI of all interrogated vessels was 0.726 with a standard deviation of 0.057. The mean RI value in the middle cerebral arteries was not significantly different with the two different measurement techniques. CONCLUSION: Normal intracranial RI values for a term infant in the first day of life were calculated for comparison with RI values in term infants with perinatal asphyxia. PMID- 11009301 TI - MR spectroscopic imaging of normal-appearing white matter in adrenoleukodystrophy. AB - BACKGROUND: Adrenoleukodystrophy (ALD) is characterised by wide phenotypic variation, and there is no marker to predict the onset of cerebral demyelination. The indications for therapeutic approaches depend largely on the onset of cerebral demyelination. OBJECTIVE: To evaluate the brain spectroscopic pattern in normal-appearing white matter (NAWM) in patients with various phenotypes of ALD to determine if these abnormalities could be of useful prognostic value. MATERIALS AND METHODS: Spectroscopic imaging acquisition mode (MRSI, 16 x 16 voxels) was performed in 20 patients with ALD, including 7 neurologically asymptomatic patients without detectable demyelination on MRI, 3 patients with early signs of cerebral demyelination, 8 patients with adrenomyeloneuropathy (AMN) and 2 patients with cerebral ALD who had previously undergone bone marrow transplantation. Controls were 22 healthy subjects. In all patients, four voxels entirely located in the juxtaventricular NAWM were studied. The ratios NAA/Cho, NAA/ CPC and Cho/CPC for the four ROIs were measured in the patient population and compared with control values. Results. In spite of a large distribution of ratios, the statistical tests did not show any significant difference between the ratios within NAWM in the patient population compared with control values. Means of ratios in the left posterior (LP) voxel compared normal subjects were (a) in neurologically asymptomatic ALD patients (n = 7) 2.27 +/- 0.63 for NAA/CPC, 2.21 +/- 0.75 for NAA/ Cho, 1.06 +/- 0.28 for Cho/CPC, (b) in patients with early signs of demyelination (n = 3) 3.43 +/- 0.85 for NAA/ CPC, 2.47 +/- 0.32 for NAA/Cho, 1.37 +/- 0.16 for Cho/CPC and (c) in AMN patients (n = 8) 1.47 +/- 0.53 for NAA/CPC, 2.17 +/- 1.58 for NAA/ Cho, 0.83 +/- 0.32 for Cho/CPC. Conclusions. The study did not show significant differences in metabolite ratios between patients and controls. The large distribution of results precludes the possibility of detecting small variations. Part of this distribution can be due to the CSI method. Longitudinal spectroscopic studies, preferentially using monovoxel spectroscopy, are clearly needed. PMID- 11009302 TI - Intrauterine subdural hematoma. AB - A patient with neonatal macrocephaly due to bilateral chronic subdural hematoma is presented. There was no history of intrauterine trauma or coagulopathy. Such patients are apparently rare. The pathogenesis of intrauterine chronic subdural hematoma in such patients is unclear. PMID- 11009303 TI - CT-guided radiolabelled aerosol studies for assessing pulmonary impairment in children with bronchiectasis. AB - OBJECTIVE: To determine whether CT-guided mucociliary clearance studies allow differentiation between bronchiectasis associated with primary ciliary dyskinesia (PCD) and those unrelated to congenital or genetically transmitted defects. MATERIALS AND METHODS: Fifteen children aged 4-18 years with a CT diagnosis of bronchiectasis were included in the study. Six had PCD, while in nine cases no congenital disorder was demonstrated. RESULTS: CT showed bronchiectasis in 26 (29%) of 90 lung regions. Radiolabelled aerosol studies were conducted globally for each lung and on the regions affected by bronchiectasis. Global half-time of activity (t 1/2) values of patients with PCD were significantly higher (P < 0.001) than those with bronchiectasis unrelated to congenital disorders. Among the 26 lung regions in which CT demonstrated bronchiectasis, regional clearance was abnormal in 24 cases. Patients with PCD showed no statistically significant difference between regional and global t 1/2 values. Patients with bronchiectasis unrelated to congenital disorders showed significantly higher regional t 1/2 values in the affected regions with respect to the corresponding global pulmonary t 1/2 (P < 0.06). CONCLUSIONS: The combination of morphological CT information with functional data concerning the clearance of radiolabelled aerosol adds to our understanding of pulmonary impairment in children with bronchiectasis. In particular, regional studies allow the recognition of different mucociliary clearance patterns in bronchiectasis associated with PCD and those unrelated to congenital or genetically transmitted defects. PMID- 11009304 TI - MR imaging of pentalogy of Cantrell variant with an intact diaphragm and pericardium. AB - We present a case of a neonate with the stigmata for pentalogy of Cantrell with the exception of diaphragmatic and pericardial defects. Diagnosing most of the anomalies in this rare syndrome can be accomplished using conventional modalities in radiology, but difficulties may arise determining diaphragmatic continuity. Accurate, early diagnosis of components of this syndrome is vital for surgical planning and assessing prognostic factors. PMID- 11009305 TI - Regional lymphadenopathy in cat-scratch disease: ultrasonographic findings. AB - BACKGROUND: Cat-scratch disease (CSD) is considered to be an emerging disease worldwide and is caused by Bartonella henselae, a gram-negative bacterium introduced by a scratch or bite of a cat. The most common clinical manifestation is regional lymphadenopathy, but clinical recognition may be difficult, as atypical manifestations may occur. The diagnosis is confirmed with serologic testing and histology is rarely needed. This paper is based on our experience with the use of ultrasonography in the diagnosis of CSD. OBJECTIVE: The aim of this study was to describe the sonographic and color Doppler appearances of regional lymphadenopathy in CSD, as this has not widely reported in the literature. MATERIALS AND METHODS: Forty-seven patients (average 9.4 years) were included who all had serologically and/or histologically proven CSD and had been studied using US early in the clinical course. All had a positive history of exposure to cats and exhibited regional lymphadenopathy. RESULTS: US showed large hypoechoic adenopathy with some transmission enhancement and high vascularization on color-flow Doppler imaging. In 30 patients, abdominal US was also performed and splenic and/or hepatic granulomata were found in 10. CONCLUSIONS: In our experience, sonography and especially color-Doppler and power-Doppler sonography was helpful in the diagnosis of CSD. We believe it should be used in the initial study of children with regional lymphadenopathy, and serologic testing should be performed when CSD is suspected. PMID- 11009306 TI - Cephaloskeletal dysplasia (Taybi-Linder syndrome: osteodysplastic primordial dwarfism type III): report of two cases and review of the literature. AB - We report two unrelated infants with cephaloskeletal dysplasia or Taybi-Linder syndrome, also referred to as osteodysplastic primordial dwarfism Type III. They presented with peculiar facial features, microcephaly and skeletal and cerebral abnormalities documented radiographically and with cranial MRI and/or CT. Some dissimilarities were observed in the skeletal findings between the two patients, most likely reflecting phenotypic variability within the same disorder. Some radiographic features were shown to evolve with time in both patients. Also of interest is the unusually long survival of these patients, more than 4 years in the first and of over 6 years in the second. PMID- 11009307 TI - Feeding intolerance secondary to marked hepatosplenomegaly compressing the stomach in children. PMID- 11009308 TI - The role of sonography in children with abdominal pain after recent successful reduction of intussusception. PMID- 11009309 TI - Teratoid Wilms' tumour occurring synchronously with classical Wilms' tumour in Beckwith Wiedemann syndrome. PMID- 11009310 TI - The twenty-five-year history of the Castroviejo Cornea Society. PMID- 11009311 TI - Ramon Castroviejo 1904-1987. PMID- 11009312 TI - Techniques in penetrating keratoplasty: a quarter century of development. PMID- 11009313 TI - The evolution of lamellar grafting techniques over twenty-five years. AB - The history of lamellar keratoplasty (LK) surgery spans over 100 years, and the advantages of lamellar surgery have long been known. The surgery is usually used for tectonic purposes; however, new techniques and technology over the past 25 years have expanded the applications of LK in optical rehabilitation. Instrumentation such as viscoelastics, diamond knives, ultrasonic pachymetry, artificial anterior chambers, advanced microkeratomes, and the excimer laser have enhanced our ability to work more safely in the tedious microsurgical environment of the lamellar procedure. Advances in surgical techniques such as deep lamellar anterior keratoplasty and deep lamellar endothelial keratoplasty have expanded the application of lamellar surgery to endothelial replacement and have achieved visual results approaching those of penetrating keratoplasty while reducing the rate of rejection and improving the long-term graft stability. As research continues, LK promises to be an increasingly important option for the corneal surgeon. PMID- 11009314 TI - The management of corneal trauma: advances in the past twenty-five years. AB - Over the past quarter century, advances in our understanding of corneal anatomy, physiology, and wound healing have all played an integral role in the management of corneal trauma. As the etiologies of corneal trauma have changed, so has our understanding of the impact of injury on corneal function as it relates to visual rehabilitation. Numerous new classes of antibiotics, antiinflammatory agents, and tissue adhesives have emerged. Occlusive therapy has advanced from simple pressure patching bandage soft contact lenses and collagen shields. Surgical instrumentation, operating microscopes, viscoelastic substances, and suture materials have all improved the outcomes of corneal trauma repair. Improved understanding of the refractive properties of the cornea through topography and alternative suture techniques has helped us restore the natural corneal curvature and visual outcomes. Consequently, in the last quarter of this century our therapeutic approaches to cornea trauma, both medical and surgical, have improved. PMID- 11009315 TI - Twenty-five-year panorama of corneal immunology: emerging concepts in the immunopathogenesis of microbial keratitis, peripheral ulcerative keratitis, and corneal transplant rejection. AB - PURPOSE: To describe the most recent advances in our understanding of the cellular and molecular mechanisms involved in the immunopathogenesis of corneal immunoinflammatory disorders including microbial keratitis, peripheral ulcerative keratitis. and allograft rejection. METHODS: Review of the published peer reviewed literature that has contributed significantly to our modern understanding of corneal immunology. In addition, the authors have summarized the information in conceptual diagrams that highlight the critical cellular and molecular pathways that lead to corneal immune responses in the two most thoroughly studied corneal immune disorders, herpes simplex keratitis (HSK) and transplant rejection. RESULTS: In spite of the wide array of molecular and cellular factors that mediate corneal immunity, critical mechanistic facets are shared by the various corneal immunoinflammatory disorders. These include activation and migration of local antigen-presenting cells (APCs), including Langerhans cells (LCs), upregulation in pleiotropic proinflammatory cytokines such as interleukin-1 (IL-1) and tumor necrosis factor-alfa (TNF-alpha) that can mediate a wide array of immune functions in addition to up-regulating protease expression. and chemokines that play a critical role on the one hand in attracting nonantigen-specific inflammatory cells such as neutrophils and on the other in attracting CD4+ T helper type 1 (Th1) cells that mediate most of the destruction in the cornea. CONCLUSIONS: In the last 25 years, we have seen our field develop from a descriptive stage into a new phase where the fundamental processes that mediate and effect corneal immunity are being accurately deciphered. It is anticipated that this new knowledge will allow development of specific molecular and genetic therapeutic strategies that could target critical steps in the immunopathogenesis of disease without the untoward side-effects of nonspecific generalized immune suppression that still remains the standard of care today. PMID- 11009316 TI - The diagnosis and management of dry eye: a twenty-five-year review. AB - PURPOSE: To review the advances in the diagnosis, pathogenesis, and management of dry eye disease in the past 25 years. METHODS: Literature review. RESULTS: The preocular tear film is a hydrated mucus gel that contains soluble antimicrobial proteins and growth factors that protect and support the ocular surface. The final common pathway in dry eye is a perturbation of the integrated ocular surface/lacrimal gland reflex unit. Diagnostic tests evaluating tear composition and clearance appear to show stronger correlation with the severity of ocular irritation symptoms and keratoconjunctivitis sicca (KCS) than the conventional Schirmer tests. KCS is a condition of abnormal differentiation and mucus production by the ocular surface epithelium that results in a poorly lubricated, abnormally permeable ocular surface that has increased susceptibility to environmental insults. Chronic subclinical ocular surface inflammation appears to play a key role in the pathogenesis of KCS. New therapeutic strategies are aimed at reducing the ocular surface inflammation of dry eye disease. CONCLUSIONS: There has been a tremendous increase in knowledge regarding dry eye disease in the past 25 years that has resulted in improved diagnostic classification and new targeted therapies. PMID- 11009317 TI - Changing concepts in the diagnosis and management of blepharitis. AB - PURPOSE: Chronic blepharitis is one of the most common conditions seen in the ophthalmologist's office; but, it is difficult to treat effectively. Nevertheless, much progress has been made over the last quarter century, not only in diagnosis but also in treatment of blepharitis. METHODS: Perhaps the most important progress has been made as the result of extensive and detailed clinical evaluations of patients, sometimes over extended periods of time with diverse patient populations. RESULTS: The availability of sophisticated instrumentation and laboratory techniques, as well as an array of antibiotics, has enhanced the whole picture for effective blepharitis treatment. CONCLUSION: Although much progress has been made, it is important to build on the present understanding. Most important is the continued development of targeted treatment protocols that address specific signs, as is now possible with microbial abnormalities. Progress in the understanding and treatment of relevant inflammatory processes will benefit from continuing biomedical discoveries. PMID- 11009318 TI - The evolution of antibiotic therapy for bacterial conjunctivitis and keratitis: 1970-2000. AB - The editors of this Festschrift asked us to review the use of antibiotics for the treatment of bacterial conjunctivitis and keratitis over the past 25 years, a period coinciding with the life of the Castroviejo Corneal Society. We believe it is more appropriate to begin our review in the late 1960s. about the time that experimental and clinical studies and algorithms for the clinical care derived from these studies helped shape a more rigorous approach to therapy. Those years saw the introduction of antibiotics that were adapted for ophthalmic use, many of which are still being used today. We will give more weight to our review of keratitis than conjunctivitis. PMID- 11009319 TI - The evolution of antiviral therapy for external ocular viral infections over twenty-five years. AB - PURPOSE: To review the past 25 years of the evolution of antiviral therapy for the treatment of common external ocular viral infections (herpes simplex virus type 1, varicella-zoster virus, and adenovirus). METHODS: A broad-based literature review in the fields of virology, antiviral research, and ophthalmology will be carried out. The pathogenesis of the major external ocular viral infections and history of antiviral development will be cited. Important conceptual breakthroughs as well as historical landmarks will be emphasized. RESULTS: The successful development of effective antivirals to treat the most common external ocular viral infections have dramatically reduced morbidity and sight loss. The immune pathogenesis of herpetic stromal keratitis is better understood. CONCLUSIONS: Remarkable progress in the development of antiviral therapy has occurred over the past quarter century. Future needs include improved antivirals and immunomodulators and vaccines to prevent and treat herpetic ocular infections and adenovirus keratoconjunctivitis. PMID- 11009320 TI - Advances in the management of keratomycosis and Acanthamoeba keratitis. AB - PURPOSE: In the late 1960s, the management of fungal keratitis was a serious unresolved problem. Little was known of the epidemiology of the disease, and there were no antifungal agents formulated for use in the eye. METHODS: A thorough review of the literature was done back to 1969 on clinical reports and experimental studies for keratomycosis and Acanthamoeba keratitis. RESULTS: Since 1969, through basic and clinical research, the epidemiology of the disease worldwide is better understood. One new topical antifungal agent, natamycin, with efficacy against filamentous fungi has been developed and the pharmacokinetics of topically applied antifungal drops have been explored. Progress has been slow but the prognosis for keratomycosis has immeasurably improved over the period. Acanthamoeba was first recognized as an ocular pathogen in 1973 and was the cause of an epidemic in the 1980s caused by contaminated contact lens, although other risk factors were also identified. At the onset of the epidemic, there was no known treatment. but as a result of intense research efforts, within a few years a well-defined therapeutic approach had been developed that had a significant impact on the prognosis for this initially devastating infection. For both infections, the role of corticosteroids for controlling the inflammation remains controversial, but the place for keratoplasty is now well defined. CONCLUSIONS: Although there has been steady progress in the management of both infections, continued research is the way to define more effective medical and surgical therapy. PMID- 11009321 TI - Changing concepts in the management of severe ocular surface disease over twenty five years. PMID- 11009322 TI - Genetics of the corneal dystrophies: what we have learned in the past twenty-five years. AB - PURPOSE: To indicate important changes in our understanding of the corneal dystrophies. METHODS: A review of the literature of the last quarter of a century. RESULTS: The earliest clinical classifications of the corneal dystrophies were based on the application of clinical, biological, histochemical, and ultrastructural methods. Since then, the first great impetus to our understanding has come from the application of techniques to map disorders to specific chromosome loci, using polymorphic markers. More recently, using candidate gene and related approaches, it has been possible to identify genes causing several of the corneal dystrophies and the mutations responsible for their phenotypic variation. A notable success has been to show that several important "stromal" dystrophies result from mutations in the gene beta ig-h3, which encodes for the protein keratoepithelin (beta ig-h3). CONCLUSIONS: For the corneal dystrophies, as with other inherited disorders, there is room for two sorts of classification system, one based mainly on clinical presentation and the other on an up-to-date understanding of the genetic mechanisms. They are not mutually exclusive. Some developmental corneal disorders are also discussed. PMID- 11009324 TI - Corneal topography and the new wave. PMID- 11009323 TI - Specular microscopy, confocal microscopy, and ultrasound biomicroscopy: diagnostic tools of the past quarter century. AB - PURPOSE: To identify, characterize, and illustrate the most important past and future potential contributions of specular, confocal, and ultrasound biomicroscopy to clinical diagnosis and research applications in the cornea from the past 25 years. METHODS: Specular microscopy, in vivo tandem scanning confocal microscopy (TSCM), scanning slit confocal microscopy (SSCM), and high-frequency ultrasound biomicroscopy are examined. RESULTS AND CONCLUSIONS: This review demonstrates the abilities and limitations of three powerful new in vivo imaging modalities to resolve the cellular and structural layers of the cornea temporally and spatially in three or four dimensions, (x, y, z, t). Clinical pathological processes such as inflammation. infection, wound healing, toxicity, embryonic development, differentiation, and disease, which previously could be studied only under static ex vivo conditions, can now be dynamically evaluated over time. Thus, with continued development and application in vivo, noninvasive microscopic techniques should provide exciting new insights into understanding the structure and function of not only the eye, but also other multicellular organ systems in health and disease. These new imaging paradigms are in the first rank of advances in medical science in the past quarter century. PMID- 11009325 TI - Twenty-five years of contact lenses: the impact on the cornea and ophthalmic practice. AB - PURPOSE: The history of contact lenses has occurred in the latter half of the 20th century. In particular, events in the 1970s through the 1980s related to the invention of soft, hydrogel contact lenses have revolutionized the contact lens industry and the eye care attached to it. This article recounts that history from the perspective of market forces, inventions, and discoveries about the physiologic functioning of the cornea. METHODS: The relevant literature is critically reviewed. RESULTS: Discoveries about the oxygen needs of the cornea and consumer pressure for clear, comfortable, around-the-clock vision have resulted in a history of rigid gas permeable and soft lenses that leads to today's contact lens picture. The short-term and long-term effects of chronic hypoxia and the levels of lens oxygen transmissibility necessary to avoid them have been well-described. The advent of the soft lens, followed by the "human experiment" with initial extended-wear modalities, led to the advent of the disposable soft contact lens. CONCLUSIONS: In the past 25 years, the development and wide acceptance of soft contact lenses have revolutionized the management of refractive error and corneal diseases. PMID- 11009326 TI - Advances in refractive surgery: 1975 to the present. AB - PURPOSE: To review the major advances in the field of refractive surgery occurring over the past 25 years. METHODS: Literature review. RESULTS: The major developments in refractive surgery over the past 25 years are reviewed. CONCLUSIONS: The past 25 years have witnessed great changes in refractive surgery. As a result of advancements in technology, instrumentation, and technique, we have seen improvements in the treatment of all types of ametropias. In this article, we review some of the successes and failures of the past quarter century. PMID- 11009327 TI - The past twenty-five years in eye banking. AB - PURPOSE: This review of milestones and challenges in eye banking describes efforts made by dedicated eye bankers to ensure the quality and quantity of corneal tissue for corneal blind patients over the last 25 years. METHODS: This account is the result of the recollection of the author, discussions with eye bankers, and a review of the literature, public documents, and Eye Bank Association of America (EBAA) records. RESULTS: A measure of the success of eye banking is the fact that corneal transplant surgery is now scheduled as an elective procedure. Corneal tissue has also enjoyed a stellar safety record. This is due in part to the establishment of comprehensive medical standards by the EBAA, the accreditation of eye banks, and the training and certification of eye bank technicians. CONCLUSION: Advancements in eye banking over the last 25 years have improved the quality of life of several hundred thousand corneal blind persons in this country. More efforts must be made in the future to eliminate corneal blindness internationally. PMID- 11009328 TI - Temperament and character in women with anorexia nervosa. AB - The present study examined temperament differences among anorexia nervosa (AN) subtypes and community controls, as well as the effect of body weight on personality traits in women with AN. Temperament and Character Inventory (TCI) scores were compared between 146 women with restrictor-type AN (RAN), 117 women with purging-type AN (PAN), 60 women with binge/purge-type AN (BAN), and 827 community control women (CW) obtained from an archival normative database. Women with AN scored significantly higher on harm avoidance and significantly lower on cooperativeness than CW. Subtype analyses revealed that women with RAN and PAN reported the lowest novelty seeking, RAN women the highest persistence and self directedness, and PAN women the highest harm avoidance. Body mass index had a nominal effect on subgroup differences, suggesting that personality disturbances are independent of body weight. Findings suggest that certain facets of temperament differ markedly between women with AN, regardless of diagnostic subtype, and controls. More subtle temperament and character differences that were independent of body weight emerged that distinguish among subtypes of AN. PMID- 11009329 TI - Chronic fatigue syndrome: occupation, medical utilization, and subtypes in a community-based sample. AB - Most studies of chronic fatigue syndrome (CFS) have been based on patients recruited from primary or tertiary care settings. Patients from such settings might not be typical of patients in the general population. The present investigation involved examining individuals with CFS from a community-based study. A random sample of 18,675 respondents in Chicago was first interviewed by telephone. A group of individuals with chronic fatigue accompanied by at least four minor symptoms associated with CFS were given medical and psychiatric examinations. From this sample, a physician review group diagnosed individuals with CFS. Those diagnosed with CFS were subclassified based on a variety of categories, including duration of illness, mode of illness onset, and presence or absence of a stressful life event directly preceding onset. In addition, we examined medical utilization among those diagnosed with CFS, as well as whether individuals with CFS were disproportionately represented in health care professions. Important differences emerged on measures of sociodemographics, symptoms, and functional disability. The implications of these findings and others are discussed. PMID- 11009330 TI - High risk model of threat perception in chronic pain patients: implications for primary care and chronic pain programs. AB - This study investigated the High Risk Model of Threat Perception (HRMTP) in middle-aged, urban chronic pain patients who had been referred to a secondary pain clinic after failing to respond to standard medical management. Relationships among absorption, social desirability (SD), and negative (e.g., depressive or anxious) affect were studied in 24 male and 73 female patients, (age range 22-88 years). Subjects completed the Tellegen Absorption Scale, the Marlowe-Crowne Social Desirability Scale, the Beck Depression Inventory--Second Edition, and the Beck Anxiety Inventory. The sample was significantly higher in SD and lower in absorption than normative groups. High SD patients endorsed significantly fewer items related to depression than those with low SD, but reported anxious ideation at about the same rate. These findings lend credence to the concept of chronic pain as a transduction of depressive, but not anxious, affect into somatic symptoms. PMID- 11009331 TI - Informativeness of structured diagnostic interviews in the identification of Tourette's disorder in referred youth. AB - Although specialized programs have greatly advanced the treatment of youth with Tourette's disorder (TD), not all children with TD reach such programs, raising questions as to whether TD is adequately identified outside specialized settings. There is thus a need for evidence that cases identified in the nonspecialty setting are "true cases." Because structured diagnostic interview methodology can reduce errors of omission, this approach can facilitate the identification of TD in referred youth outside specialized programs. Similarities between cases ascertained in specialty and nonspecialty settings would suggest that those identified in the nonspecialty setting were indeed "true cases." Comparisons were made between youth with TD ascertained through a specialized TD program who had both a structured diagnostic interview-derived diagnosis of TD plus an expert evaluation of TD (N = 103), with youth ascertained through a non-TD specialized pediatric psychopharmacology program who had a structured diagnostic interview derived diagnosis of TD (N = 92). Irrespective of ascertainment source, children with structured interview-derived diagnosis of TD shared similar correlates in terms of tic severity, mean age of onset and duration of tics, as well as patterns of comorbidity well known to be associated with TD in clinical samples. Children meeting diagnostic criteria for TD on structured diagnostic interviews share similarities and patterns of clinical correlates, irrespective of ascertainment through a specialized TD or non-TD specialized clinic. These findings support the usefulness of structured diagnostic interview methodology as a diagnostic aid for the identification of TD in non-TD specialized settings and facilitate delineation of patterns of comorbidity. PMID- 11009332 TI - A comparison of posttraumatic stress disorder with and without borderline personality disorder among women with a history of childhood sexual abuse: etiological and clinical characteristics. AB - The overlap in definition and presentation between posttraumatic stress disorder (PTSD) and borderline personality disorder (BPD) has raised questions about the relationship of these disorders. Are they separate disorders, variants of the same disorder, or comorbid conditions? The present study examined etiological variables and current functioning among two groups of outpatient women with a history of childhood sexual abuse: those with PTSD only (N = 45) and those with PTSD and BPD (N = 26). The groups did not differ in severity, frequency, or number of perpetrators of their childhood sexual abuse, or whether the perpetrator was a family member or not. The additional diagnosis of BPD was associated with earlier age of abuse onset and significantly higher rates of physical and verbal abuse by mother. Severity and frequency of PTSD symptoms were not affected by BPD diagnosis, suggesting that the personality disorder and PTSD are independent symptom constructs. The PTSD+BPD group scored higher on several other clinical measures including anger, dissociation, anxiety, and interpersonal problems. They did not differ in their frequency of use of mental health services but tended to be less compliant in their treatment. These and other findings are discussed, and implications for treatment are considered. PMID- 11009333 TI - Childhood emotional trauma and chronic posttraumatic stress disorder in adult outpatients with treatment-resistant depression. AB - The intent of this study was to test the hypothesis that patients with treatment resistant depression are more likely than treatment responsive patients to suffer from sequelae of childhood trauma that may perpetuate depression despite adequate medication treatment. Twenty participants with treatment-resistant depression and 20 participants with treatment-responsive depression were administered a structured interview and a battery of psychological tests to assess levels of current depression, confirm diagnosis, and quantify childhood trauma and presence of dissociative phenomena. Tests used include the Beck Depression Inventory, the Mini International Neuropsychiatric Interview, the Minnesota Multiphasic Personality Inventory-2, the Childhood Trauma Questionnaire, and the Trauma Symptom Inventory. Compared with treatment responders, the treatment-resistant participants were significantly more depressed, had significantly more comorbid anxiety disorders, reported significantly greater levels of childhood emotional abuse, and experienced current-day sequelae of childhood emotional abuse. The hypothesis was partially supported by these results. This study suggests that reported history of childhood emotional abuse and sequelae of that abuse may be associated with treatment resistance in depressed outpatients. PMID- 11009334 TI - Memory for acute stress disorder symptoms: a two-year prospective study. AB - The present study aimed to index the accuracy of memory for acute trauma symptoms by comparing the symptoms reported by motor vehicle accident (MVA) victims within 1 month posttrauma with the recall of these symptoms at 2 years posttrauma. Ninety-two consecutive MVA admissions were assessed for the presence of acute stress disorder (ASD) within 1 month posttrauma. At 2 years posttrauma, 61% (N = 56) of the sample were reassessed for posttraumatic stress disorder (PTSD) and for accuracy of recall of the symptoms reported during the first assessment. At least one of the four ASD diagnostic clusters was recalled inaccurately by 75% of patients. High levels of posttraumatic stress severity and high subjective ratings of injury severity at 2 years posttrauma were associated with errors of addition (i.e., recalling the presence of acute symptoms 2 years posttrauma that were not reported during the first assessment). Low levels of posttraumatic stress severity and low subjective ratings of injury severity at 2 years posttrauma were associated with errors of omission (i.e., omitting to recall acute symptoms 2 years posttrauma that were reported during the first assessment). These results suggest that retrospective reports of acute stress symptoms should be interpreted cautiously because of the influence of current symptoms on recall of acute symptoms. PMID- 11009335 TI - Assessing problems with religious content: a comparison of rabbis and psychologists. AB - This study measured distinctions made by a sample of clergy and mental health professionals in response to three categories of presenting problems with religious content: mental disorder, religious or spiritual problem, and "pure" religious problem. A national, random sample of rabbis (N = 111) and clinical psychologists (N = 90) provided evaluations of three vignettes: schizophrenia, mystical experience, and mourning. The participants evaluated the religious etiology, helpfulness of psychiatric medication, and seriousness of the presenting problems. The rabbis and psychologists distinguished between the three diverse categories of presenting problems and concurred in their distinctions. The results provide empirical evidence for the construct validity of the new DSM IV category religious or spiritual problem (V62.89). Use of the V code allows for more subtle distinctions among the variety of problems that persons bring to clergy and mental health professionals. These distinctions may also provide a foundation for the initiation of co-professional consultation. PMID- 11009336 TI - Religion: a mediator of Anglo-American and Mexican attributional differences toward symptoms of schizophrenia? AB - This study examined the relationship of religiosity to attributions toward schizophrenia, within a cultural context. Previous research suggests that on self report measures, Mexicans endorse holding greater moral-religious values than do their Anglo-American counterparts. Research also indicates that Mexicans, relative to Anglo-Americans, tend to hold fewer blameworthy attributions and are less likely to view patients with schizophrenia as responsible for the symptoms of the disorder. In an analog study of 88 Mexican and 88 Anglo-American college students asked to imagine that they have a brother with schizophrenia, this study assessed two competing hypotheses regarding the role of religion in shaping reactions to schizophrenia in a family member. For hypothesis 1, Baron and Kenny's mediational model (1986) was used to assess whether moral religious values may play a direct mediating role between ethnicity and controllability attributions for schizophrenia. In other words, based on observations of previous researchers, this set of analyses assessed whether ethnic differences in controllability attributions might be explained by a religious or spiritual tendency in Mexicans to view negative events, such as mental disability, as rooted in divine factors beyond the patient's personal control. In contrast, a second, competing, hypothesis was also assessed in this study: namely, that greater religiosity would be positively correlated with increasing perceptions of control over the symptoms of schizophrenia. This hypothesis stems from the premise of several investigators that religious individuals may be more likely to perceive another's adversity (such as having schizophrenia) as a punishment for prior wrongdoings or for failure to try to help oneself. Results indicate support for the latter hypothesis. Implications of the paradoxical finding, that Mexicans were both more religious and more external in their attributions, are discussed. PMID- 11009337 TI - Exhausted neurosurgeon describes a porcine brain implant to wine merchant with Parkinson's disease PMID- 11009338 TI - Trauma orientation and detection of violence histories in the psychiatric emergency service. PMID- 11009339 TI - Negative priming in schizophrenia: no evidence for reduced cognitive inhibition. PMID- 11009340 TI - Heck arylation of endocyclic enecarbamates with diazonium salts. Improvements and a concise enantioselective synthesis of (-)-codonopsinine. AB - Total enantioselective synthesis of the natural (-)-codonopsinine was accomplished in seven steps with an overall yield of approximately 16% starting from the five-membered endocyclic enecarbamate 4. The total synthesis features a highly efficient and stereoselective Heck arylation of endocyclic enecarbamate 4 with p-methoxybenzenediazonium tetrafluoroborate and a stereoselective epoxidation/epoxide opening sequence as key steps. PMID- 11009342 TI - PINDY: A novel, pinene-derived bipyridine ligand and its application in asymmetric, Copper(I)-catalyzed allylic oxidation AB - The title bipyridine ligand (+)-6(PINDY), prepared in five steps from (-)-beta pinene, forms a stable complex with CuCl(2) (8) that has been characterized by X ray crystallography to reveal an unusual geometry at Cu. Triflate 9 proved to catalyze asymmetric allylic oxidation (10 --> 11; rt, approximately 30 min, 49 75% ee). PMID- 11009341 TI - O-Glycosyl trichloroacetimidates bearing Fmoc as temporary hydroxy protecting group: a new access to solid-phase oligosaccharide synthesis. AB - Different O-glycosyl trichloroacetimidates bearing base sensitive Fmoc protected hydroxy groups were efficiently prepared with CCl(3)CN using a catalytic amount of sodium hydride. The resulting glycosyl donors were engaged in glycosylation reactions both in solution and on solid support with a new ester-type linker with good results. In both approaches, Fmoc groups were afterward quantitatively cleaved using mild basic conditions. PMID- 11009343 TI - A linear multiporphyrinic AB - A linear multiporphyrinic [2]-rotaxane has been synthesized using the transition metal-templating method for threading a gold(III)-incorporating macrocycle onto a rodlike, phenanthroline-derived chelate bearing carboxylate end groups. Stoppering has been performed by reacting the resulting prerotaxane with the amino derivative of a zinc tetraarylporphyrin under EDC-HOBt activation. A 34% yield has been realized for this one-pot, double amide bond formation. PMID- 11009344 TI - Complete N-1 regiocontrol in the formation of N-arylimidazoles. Synthesis of the active site His-Tyr side chain coupled dipeptide of cytochrome c oxidase. AB - Under catalysis by copper(II) acetate, complete regiocontrol (N-1 versus N-3) was obtained in the arylation of substituted imidazoles with aryllead(IV) reagents. The mildness of the reaction conditions (rt, no added base) allows for the first synthesis of the histidine-tyrosine side chain coupled dipeptide found in the active site of cytochrome c oxidase. PMID- 11009345 TI - A DFT and PM3 study of a hypothetical silicon-based sigma-allyl cation and sigma allyl radical and higher homologues AB - DFT and PM3 calculations were used to model the hypothetical sigma-allyl cation 1 Si(3)()(+) and sigma-allyl radical 1-Si(3)()(*). For both the cation and the radical, the structure with equal Si-Si bond lengths and sigma-delocalized charge or spin is predicted to be favored relative to a bond-localized "capped" triquinane alternative. Higher homologues of 1-Si(3)()(+) and 1-Si(3)()(*) were also modeled; they likewise are predicted to prefer sigma-delocalized structures. PMID- 11009346 TI - Conformational analysis of alkylated biuret and triuret: evidence for helicity and helical inversion in oligoisocyanates AB - The conformations of several oligoisocyanates have been investigated by NMR in order to study the onset and dynamics of helicity in polyisocyanates. Pentaethylbiuret and hexaethyltriuret were found to adopt turns and helices in solution. For hexaethyltriuret, symmetric and asymmetric helices are present. Not only is an interconversion of these forms observed (DeltaG(SA)(double dagger) = 9.3 +/- 0.4 kcal/mol) but also a reversal of helicity (DeltaG(PM)(double dagger) = 9.0 +/- 0.4 kcal/mol). The coalescence pattern for the latter process provides direct evidence for a concerted, conrotatory helical inversion. PMID- 11009348 TI - Fragmentation reaction via an ipso-attack in the multistep synthesis of regioselectively functionalized Calix AB - The acid-catalyzed convergent synthesis of a calix[8]arene containing a regioselectively functionalized upper rim has been investigated through the isolation and identification of cyclic and acyclic byproducts. Theoretical calculations demonstrated that the synthetic process involves two types of reaction mechanisms, one of which leads to the favorably constructed framework, while the other causes undesirable fragmentation via ipso-substitution. A possible rationale is proposed to explain the overall reaction pathways which derive the calix[8]arene along with the byproducts. PMID- 11009347 TI - Polymerized fluorescent liposomes incorporating lanthanide ions. AB - This paper describes the design and synthesis of a polymerizable lipid capable of complexing lanthanide ions. The lipid has been successfully incorporated into liposomes and then polymerized. Fluorescence studies indicate that the diacetylene (unpolymerized lipid) and the conjugated alkenes (after polymerization) can be used as sensitizers for the lanthanide ion. PMID- 11009349 TI - Solid-phase synthesis of 1,2,3, 4-tetrahydro-beta-carboline-containing peptidomimetics. AB - A solid-phase method for the synthesis of 1,2,3, 4-tetrahydro-beta-carboline containing peptidomimetics has been developed. The key step in the strategy is the Pictet-Spengler condensation of a resin-bound tryptophan-containing fragment with an Fmoc-amino aldehyde. PMID- 11009350 TI - Novel phenylazomethine dendrimers: synthesis and structural properties AB - The novel dendrimers consisting of a pi-conjugated backbone, the dendritic polyphenylazomethines (DPAs), were synthesized by the convergent method via the dehydration of aromatic ketones with aromatic amines in the presence of titanium(IV) tetrachloride. The obtained DPAs have a high solubility unlike the conventional linear polyphenylazomethines. NMR studies revealed the conformational rigidity of DPA G4. DPAs having many azomethine groups as the coordination site for metal ions are expected to be novel ligands. PMID- 11009351 TI - Diastereoselective intramolecular Ritter reaction: generation of a cis-fused hexahydro-4aH-indeno[1,2-b]pyridine ring system with 4a,9b-diangular substituents. AB - Indanol intermediates 5, prepared via Michael addition of 1-indanone beta ketoester and acrylonitrile followed by reduction or Grignard reaction of the ketone group, were submitted to intramolecular Ritter reaction using various acid reaction conditions to produce tricyclic lactams 4. This cis-fused hexahydro-4aH indeno[1,2-b]pyridine ring system, substituted at both angular positions 4a and 9b, provides access to constrained analogues of non-peptide NK(1)-antagonists with monocyclic piperidine structure. PMID- 11009352 TI - Dipolar cycloaddition of novel 6-(nitrileoxidomethyl) penam sulfone: an efficient route to a new class of beta-lactamase inhibitors. AB - 6-(Nitrileoxidomethyl) penam sulfone intermediate was prepared in a few steps starting from commercially available (+)-6-aminopenicillanic acid. This intermediate underwent smooth 1, 3-dipolar cycloaddition reactions with various alkenes and alkynes to give cycloadducts in moderate to good yields. By this new method, several potent beta-lactamase inhibitors were synthesized. The regio- and stereoselectivity outcomes of the cycloaddition process are also discussed. PMID- 11009353 TI - Facile, completely regioselective 1,4-hydrogenations of C(60)-diaryltetrazine monoadducts AB - Thermal Diels-Alder reactions between C(60) and electron-deficient 3, 6-diaryl 1,2,4,5-tetrazines yield monoadducts possessing a diaryldihydropyridazine function nested atop the fullerene. The diaryldihydropyridazine functions direct a completely regioselective 1,4-hydrogenation, resulting in a racemic mixture of bifunctional bisadduct (+/-)-3. PMID- 11009354 TI - 1-Aminocyclopropaneboronic acid: synthesis and incorporation into an inhibitor of hepatitis C virus NS3 protease. AB - The previously unreported alpha,alpha-disubstituted 1-aminoboronate esters have potential utility in peptidomimetic design, particularly against serine protease targets. A concise synthesis of 1-aminocyclopropaneboronate pinanediol ester is reported, and a peptidyl derivative is shown to have modest affinity (K(i) = 1.6 microM) for hepatitis C NS3 protease. PMID- 11009355 TI - Effect of competing alkali metal cations on neutral Host's anion binding ability AB - Anion binding by neutral hosts in organic solvents can be inhibited by the presence of alkali metal cations. The binding inhibition is due to salt ion pairing which increases in the order Cs(+) < K(+) < Na(+). The binding inhibition can be reversed by using heteroditopic hosts that simultaneously bind both the metal cation and the anion. The largest cation-induced enhancements are observed with the less basic anions. PMID- 11009356 TI - Novel synthesis of hybrid calixphyrin macrocycles AB - The reaction of benzaldehyde with excess pyrrole at room temperature in the absence of solvent affords a mixture of meso-substituted polypyrranes species. After separation by column chromatography, these may be used to prepare a range of calix[4]phyrin macrocycles by condensation with acetone under conditions of acid catalysis. PMID- 11009357 TI - Regioselective synthesis of unsymmetrical 3, 5-dialkyl-1-arylpyrazoles AB - 3-Alkoxylmethyl-5-alkylpyrazoles undergo regioselective N-arylation with 4 fluoronitrobenzene in the presence of base to yield the corresponding 1-(4-nitro phenyl)pyrazoles. Further elaboration of these intermediates furnishes a practical synthesis of unsymmetrical 3,5-dialkyl-1-arypyrazoles. A tentative explanation of the observed regioselectivities is provided. PMID- 11009358 TI - Polymer support assisted selective functionalization of azoles using a palladium catalyzed coupling reaction AB - Regioselective palladium-catalyzed monoarylation of azoles was achieved using iodobenzoate immobilized on an insoluble polymer support. The positional selectivity for the coupling reaction was dramatically influenced by the presence of CuI. No diarylation was observed in either case. Unsymmetrical diarylation was also obtained with the polymer support using sequential coupling reactions. PMID- 11009359 TI - Convenient route to super-expanded calixpyrroles: synthesis of Calix AB - meso-Alkylporphyrinogen-like cyclic oligomers containing furans and pyrroles have been synthesized by "3 + 2" and "4 + 2" approaches. Condensation of 5,5,10,10,15,15-hexaethyl-21, 22-dioxatetrapyrromethane with 2,5-bis[(alpha hydroxy-alpha, alpha-dimethyl)furan] resulted in the formation of cyclic hexamer and cyclic dodcamer. Effects of catalysts, temperature, inorganic additives, solvent, and reaction concentration were examined. PMID- 11009361 TI - Radical-initiated, skeletal rearrangements of Bicyclo AB - Treatment of bicyclic lactones derived from Diels-Alder reactions of 3 carbomethoxy-2-pyrone under radical conditions leads to a series of interesting skeletal rearrangements. The stereochemical and optical integrity of the starting material are maintained throughout the process. PMID- 11009360 TI - An efficient chiral moderator prepared from inexpensive (+)-3-carene: synthesis of the HIV-1 non-nucleoside reverse transcriptase inhibitor DPC 963. AB - The beta-amino alcohol 4 beta-morpholinocaran-3 alpha-ol is prepared by addition of morpholine to alpha-3,4-epoxycarane utilizing anhydrous magnesium bromide as Lewis acid promoter. The enantiopure amino alcohol is uniquely effective as a chiral moderator for the addition of lithium cyclopropylacetylide to an unprotected N-acylketimine. This reaction provides an efficient route to the second generation NNRTI drug candidate DPC 963. PMID- 11009362 TI - Efficient asymmetric synthesis of radicicol dimethyl ether: a novel application of ring-forming olefin metathesis. AB - A concise, stereospecific synthesis of radicicol dimethyl ether is presented. The strategy relies on a convergent three-stage assembly of the 14-membered lactone which has, as a key transformation, a novel ring-forming metathesis reaction utilizing a vinyl epoxide. PMID- 11009363 TI - Cobalt-catalyzed alkyne-nitrile cyclotrimerization to form pyridines in aqueous solution. AB - A new, water-soluble cobalt(I) catalyst has been used in the aqueous, chemospecific, cyclotrimerization of one nitrile with two alkynes for the synthesis of highly functionalized pyridines. Several different functional groups are well incorporated in this transformation, including unprotected alcohols, ketones, and amines. Double isotopic crossover data, as well as nitrile dependence on the rate of product formation, suggest associative rate-determining coordination of the nitrile. PMID- 11009364 TI - The mild cleavage of 2-amino-2-deoxy-D-glucoside methoxycarbonyl derivatives. AB - The conversion of methyl carbamate to the corresponding free amine is described for a series of 2-amino-2-deoxy-D-glucosamine derivatives. Cleavage of methoxycarbonyl moiety with MeSiCl(3) and triethylamine in dry THF at 60 degrees C and subsequent aqueous hydrolysis yields the free amine in 54 to 93% yields. The selective cleavage of methyl carbamates with MeSiCl(3) in the presence of a 2,2,2-trichloroethoxycarbonyl group or 2-azido glycosides affords selectively, orthogonal N-deprotected carbohydrates. PMID- 11009365 TI - Efficient synthesis of the D-ring fragment of cobyric acid AB - The synthesis of a highly functionalized 4,5-dihydro-3H-pyrrol, namely, the D ring fragment 5a of cobyric acid (1), is described in this letter. A very efficient assembly to 5a involves CBS-reduction of 10, a [2,3] Wittig-Still rearrangement, and a stereoselective Michael addition to a nitro olefin. PMID- 11009367 TI - Stereocontrol in rare earth metal triflate-catalyzed 1,3-dipolar cycloaddition reaction of 2-benzopyrylium-4-olate with aldehydes AB - Exo-cycloadduct was obtained with high stereoselectivity by the addition of ytterbium triflate (Yb(OTf)(3)) (10 mol %) in the rhodium(II) acetate (Rh(2)(OAc)(4))-catalyzed decomposition of o-(methoxycarbonyl)-alpha diazoacetophenone in the presence of benzaldehyde. In the reaction in the absence of Yb(OTf)(3), almost no selectivity was obtained. Stereoselectivity in the reactions with p-nitrobenzaldehyde, p-anisaldehyde, and benzyloxyacetaldehyde could be also controlled to high exo preference by the addition of Yb(OTf)(3). PMID- 11009366 TI - Reaction of COTC with glutathione: structure of the putative glyoxalase I inhibitor. AB - The structure of the active glyoxalase I inhibitor derived from the Streptomyces griseosporeus metabolite COTC 1 has been conclusively identified by means of total synthesis as 2c. Human glyoxalase I is competitively inhibited by 2c (K(i)() = 183 +/- 6 microM) but is not inhibited by 1 itself. PMID- 11009368 TI - A novel strategy for designing irreversible inhibitors of metalloproteases: acetals as latent electrophiles that interact with catalytic nucleophile at the active site. AB - A new strategy for design of irreversible inactivators for carboxypeptidase A (CPA), a prototypic zinc protease, has been developed by exploiting the property of acetals to generate an oxacarbenium ion intermediate in the conversion into the corresponding carbonyl compounds. The design strategy is exemplified by 2 benzyl-5-alkyl-3,5-dioxapentanoic acids (1a-c). Interestingly, (R)-1b is slightly more potent than an (S)-1b as an inactivator of CPA. PMID- 11009369 TI - In situ preparation of a highly active N-heterocyclic carbene-coordinated olefin metathesis catalyst. AB - Highly active N-heterocyclic carbene-coordinated catalysts may be synthesized and used in situ, without requiring prior isolation of the catalyst. Activation of this in situ catalyst with ethereal HCl dramatically reduces the reaction times required for high conversions. A variety of alpha,beta-unsaturated carbonyl containing substrates participate readily in cross and ring-closing metathesis reactions using this preparation. PMID- 11009370 TI - Highly enantioselective addition of diethylzinc to diphenylphosphinoyl imines under dual amino Alcohol/Halosilane mediation AB - Arylethylene-derived, enantiomerically pure amino alcohols have been evaluated as ligands for the dual-catalyzed (amino alcohol/halosilane) enantioselective addition of diethylzinc to diphenylphosphinoyl imines. Among them, the conformationally restricted 9-fluorenone-derived ligand 4c provides the highest enantioselectivities so far reported over a range of substrate imines. PMID- 11009371 TI - A synthetic transmembrane polyether model active in lipid bilayers. AB - A model polyether-based ion channel-like compound was rationally designed and synthesized. Macromolecules 1c-f were incorporated into phospholipid vesicles and shown to facilitate the transmembrane sodium transport. PMID- 11009373 TI - Chiral discrimination in hydrogen-bonded AB - A series of racemic [7]helicenes have been prepared and characterized both in solution and in the solid state. Despite the helicenes having the ability to self assemble in a variety of stereochemical and topological relationships, they formed only enantiomerically pure dimers held together by two pairs of cooperative hydrogen bonds. The self-assembly process was enantiospecific in solution and diastereoselective in the crystal. PMID- 11009372 TI - First asymmetric hetero Diels-Alder reaction of 1-sulfinyl dienes with nitroso derivatives. A new entry to the synthesis of optically pure 1,4-imino-L-ribitol derivatives. AB - Hetero Diels-Alder (HDA) cycloaddition of chiral 1-p-tolylsulfinyl-1,3-pentadiene with benzyl nitrosoformate, under mild conditions, yields 2H-1,2-oxazine 3 with complete regioselectivity and pi-facial diastereoselectivity. Sequential osmylation and protection of the resulting glycol gives the oxazine 5 which is directly transformed into enantiomerically pure 1,4,5-trideoxy-1,4-imino-L ribitol 8 by reduction under Pd/C. PMID- 11009374 TI - A facile stereocontrolled synthesis of anti-alpha-(trifluoromethyl)-beta-amino alcohols. AB - A short stereocontrolled preparation of anti-alpha-(trifluoromethyl)-beta-amino alcohols is described, involving an initial CF(3) transfer to cinnamaldehyde and a one-step, three-component condensation of 3,3,3-trifluorolactic aldehyde, an alkenyl (aryl) boronic acid, and an amine. Applying this methodology to chiral 3,3,3-trifluorolactic aldehyde allowed us to generate an amino alcohol enantioselectively in 92% ee. PMID- 11009375 TI - Novel total synthesis of the anticancer natural product dysidiolide. AB - The marine natural product dysidiolide has been synthesized in a highly diastereoselective fashion that features the sequential transfer of chirality from a cyclohexenone precursor. PMID- 11009376 TI - A concise route to (-)-kainic acid. AB - A concise route to (-)-kainic acid from enantiopure (+)-cis-4-carbobenzoxyamino-2 cyclopentenol has been devised by employing concurrent Chugaev syn-elimination and intramolecular ene reaction as the key step. PMID- 11009377 TI - A facile synthesis of dragmacidin B and 2,5-bis(6'-bromo-3'-indolyl)piperazine. AB - A short synthesis of dragmacidin B (1), 2,5-bis(6'-bromo-3'-indolyl)piperazine (2), and corresponding didebromo analogues 8 and 9 is described. The key steps involve the dimerization of oxotryptamines 4 and 11 to give bis(indolyl)pyrazines 5 and 12, which upon selective reduction and reductive methylation with sodium cyanoborohydride afforded the requisite piperazine natural products. PMID- 11009378 TI - Synthesis of novel acetylenic cyclophanes with helical chirality: potential new structures for liquid crystals AB - The synthesis of a series of novel acetylenic cyclophanes is described. X-ray crystallographic analysis of the core structure revealed a twisted conformation with helical chirality. Preliminary results suggest that these cyclophanes, with appropriate functionality, have the potential to act as unique liquid crystalline materials. PMID- 11009379 TI - A diastereocontrolled synthesis of (+)-febrifugine: a potent antimalarial piperidine alkaloid. AB - A diastereocontrolled synthesis of (+)-febrifugine, a potent antimalarial piperidine alkaloid, has been achieved using a chiral block having a bicyclo[3.2.1]octane framework which exhibits inherent convex-face selectivity. PMID- 11009380 TI - Yb(OTf)(3)-Catalyzed oxymercuration of homoallylic alcohol-derived hemiacetals and hemiketals. AB - 1,3-Diol synthons, protected as acetonides or benzylidene acetals, may be synthesized efficiently from homoallylic alcohols and acetone or benzaldehyde by oxymercuration of the derived hemiketals and hemiacetals with HgClOAc. The use of catalytic amounts of Yb(OTf)(3) is crucial to the success of the reaction, which was determined to be reversible. PMID- 11009381 TI - Convenient iterative synthesis of an octameric tetracarboxylate-functionalized oligophenylene rod with divergent end groups AB - Oligo(p-phenylene) rigid rod 10 is synthesized via a functional group-tolerant molecular doubling approach. Preparative chromatographic methods, protecting groups, boronic acid isolations, and Grignard or organolithium reagents are not used. The convenient synthesis of well-defined, polar-functionalized oligophenylene rigid rods could afford ready access to a variety of useful electronic organic materials. PMID- 11009383 TI - An expeditious route to eight- and nine-membered carbocycles based on a RCM-ring fragmentation strategy AB - The presence of a temporary one-atom internal tether in 1,9-deca- and 1,10 undecadienes allows their efficient ring-closing metathesis (RCM). Cleavage of the bridging tether of the resulting bicycles provides eight- or nine-membered carbocycles, medium-sized rings that are difficult to assemble using other currently available procedures. PMID- 11009382 TI - Rhodium-catalyzed formylation of organomercurials: application to efficient polyol synthesis. AB - The rhodium-catalyzed formylation of organomercurials-a new transformation of organomercurials-is reported. The addition of 0.50 equiv of 1,4 diazabicyclo[2.2.2]octane (DABCO) was found to promote the reaction, and it is postulated that the DABCO acts as a ligand for mercury. Several examples are presented to document the scope of the reaction. This reaction was developed in the context of a larger program focused on the development of efficient strategies for the synthesis of polyol-derived natural products, and an efficient (8 steps) synthesis of Tolypothrix pentaether that employs this methodology is reported. PMID- 11009384 TI - Phosphorus pentasulfide: A mild and versatile Catalyst/Reagent for the preparation of dithiocarboxylic esters AB - The reaction of carboxylic acids (1) with a variety of thiols or alcohols in the presence of phosphorus pentasulfide (P(4)S(10)) as a catalyst and reagent (20-40 mol %) proceeded effectively to afford the corresponding dithiocarboxylic esters (2) in high yields. PMID- 11009385 TI - Construction of 4-hydroxy-2-pyridinones. Total synthesis of (+)-sambutoxin. AB - Total synthesis of (+)-sambutoxin has been achieved, establishing the relative and absolute stereochemistry of the naturally occurring mycotoxin. Efforts feature methodology for enantiocontrolled construction of 1,3-anti-dimethyl arrays and a novel Saegusa oxidation to provide a pyridinone methide leading to formation of the central dihydropyran ring. PMID- 11009386 TI - Mild and general cross-coupling of (alpha-Alkoxyvinyl)silanols and -silyl hydrides AB - (alpha-Alkoxyvinyl)silanols and (alpha-alkoxyvinyl)silyl hydrides are efficiently converted to aryl vinyl ethers by a palladium(0)-catalyzed cross-coupling reaction with aryl halides in the presence of tetrabutylammonium fluoride or hydroxide. Yields are generally high, and the reaction is compatible with a wide range of functional groups. PMID- 11009387 TI - Triple hydroxylation of tetracenomycin A2 to tetracenomycin C involving two molecules of O(2) and one molecule of H(2)O. AB - The TcmG or ElmG oxygenase-catalyzed triple hydroxylation of tetracenomycin (Tcm) A2 to Tcm C proceeds via a novel monooxygenase-dioxygenase mechanism, deriving the 4- and 12a-OH groups of Tcm C from two molecules of O(2) and the 4a-OH group of Tcm C from a molecule of H(2)O. These results suggest a mechanistic analogy among TcmG, ElmG, and the bacterial and fungal hydroquinone epoxidizing dioxygenases, as well as the mammalian vitamin K-dependent gamma-glutamyl carboxylase. PMID- 11009388 TI - Thiol ester-boronic acid cross-coupling. Catalysis using alkylative activation of the palladium thiolate intermediate. AB - Thiol esters and boronic acids do not participate in cross-coupling in the presence of palladium catalysts. However, efficient palladium-catalyzed thiol ester-boronic acid cross-coupling is observed when simple alkylating agents are present. Alkylative conversion of the very stable palladium-thiolate bond to a labile palladium-thioether bond is presumed to be crucial to the catalysis. Of the systems studied, 4-halo-n-butyl thiol esters were most effective in this cross-coupling. PMID- 11009389 TI - Formal synthesis of (+/-)-dendrobine: use of the amidofuran cycloaddition/rearrangement sequence. AB - The formal synthesis of the alkaloid (+/-)-dendrobine (4) was accomplished using the IMDAF cycloaddition/rearrangement sequence of a furanyl carbamate. Conversion of the rearranged cycloadduct to Kende's advanced intermediate in eight steps completed the formal synthesis of (+/-)-dendrobine. PMID- 11009390 TI - A general synthetic method for the formation of substituted 5-aminotetrazoles from thioureas: a strategy for diversity amplification. AB - A general method for the synthesis of 5-aminotetrazoles is outlined using the mercury(II)-promoted attack of azide anion on a thiourea. The reaction proceeds through a guanyl azide intermediate, which undergoes electrocyclization to the tetrazole. The method is high yielding and provides access to mono-, di-, and trisubstituted 5-aminotetrazoles, targets of potential interest for combinatorial library development. PMID- 11009391 TI - Investigation of a dialkylation approach for enantioselective construction of vicinal quaternary stereocenters. AB - A detailed study of the dialkylation of dianions derived from dihydroisoindigo 1 with enantiopure ditriflate 2 is reported. The LHMDS-mediated process has been optimized to give C(2)-symmetric product 3 with high selectivity (C(2) selectivity 3:5 = 100:1; C(2):C(1) selectivity = 8:1). Stereoselection in the C(2) manifold is determined in both the bimolecular and intramolecular alkylation steps. PMID- 11009392 TI - Synthesis of carbapenam skeletons using a ruthenium-catalyzed cyclization. AB - Carbapenam is a very important skeleton of beta-lactam antibiotics, and it has a highly strained structure. When enynes 9 were treated with RuH(2)CO(PPh(3))(3) (10 mol %) in toluene upon heating, carbapenams 10 were obtained in good yields. PMID- 11009393 TI - The principle of laplace and scaling of ventricular wall stress and blood pressure in mammals and birds. AB - Maximum left ventricular wall stress is calculated at end-diastolic volume and systemic arterial diastolic blood pressure, according to a thick-walled model for the principle of Laplace. Stress is independent of body mass and averages 13.9 kPa (+/-2.3; 95% confidence interval) in 24 species of mammals weighing 0.025 4,000 kg and 15.5 kPa (+/-4.7) in 12 birds weighing 0.014-110 kg. Birds have higher arterial blood pressures and larger hearts than mammals. Systolic and diastolic arterial blood pressures increase with body mass according to M(0.05) in mammals, and heart mass increases according to M(1.06) in the same species, further supporting the principle. However, blood pressure in birds is independent of body mass, and heart mass scales isometrically. End-diastolic stress values, calculated according to Laplace, are about one-third of peak stresses recorded in isolated mammalian myocardial preparations. PMID- 11009394 TI - Cold tolerance and the regulation of cardiac performance and hemolymph distribution in Maja squinado (Crustacea: decapoda). AB - Elevated Mg(2+) levels in the hemolymph ([Mg(2+)](HL)) of brachyuran crabs have recently been demonstrated to limit cold tolerance by reducing motor and circulatory activity. Therefore, the limiting function of elevated [Mg(2+)](HL) on circulatory performance and arterial hemolymph flow was investigated by the pulsed-Doppler technique in the spider crab Maja squinado during progressive cooling from 12 degrees to 0 degrees C. [Mg(2+)](HL) were reduced from control levels of 39.9 mmol L(-1) to levels of 6.1 mmol L(-1) by incubation in magnesium reduced seawater. At 12 degrees C cardiac output was 13.9+/-2.4 mL kg(-1) min(-1) and stroke volume 0.2+/-0.04 mL kg(-1) min(-1) in control animals. In [Mg(2+)](HL)-reduced animals cardiac output increased to 43.6+/-5.0 mL kg(-1) min(-1) and stroke volume rose to 0.6+/-0.1 mL kg(-1) min(-1). Temperature reduction in control animals revealed a break point at 8 degrees C linked to a major redirection of hemolymph flow from lateral to sternal and hepatic arteries. Cardiac output and heart rate dropped sharply during cooling until transiently constant values were reached. Further heart rate reduction occurred below 4.5 degrees C. Such a plateau was not detected in [Mg(2+)](HL)-reduced animals where the break point decreased to 6 degrees C, also indicated by a sharp drop in heart rate and cardiac output and the redirection of hemolymph flow. It is concluded that progressive cooling brings the animals from a temperature range of optimum cardiac performance into a deleterious range when aerobic scope for activity falls before critical temperatures are reached. Reduction of [Mg(2+)](HL) shifts this transition to lower temperatures. These findings support a limiting role for [Mg(2+)](HL) in thermal tolerance. PMID- 11009395 TI - Effects of food shortage and oversupply on energy utilization, histology, and function of the gut in nestling song thrushes (Turdus philomelos). AB - We measured food intake, digestive efficiency, body mass increments, resting metabolic rate (RMR), carcass fat content, size and histological structure of the gut, and the rate of intestinal brush border uptake of l-proline in song thrush (Turdus philomelos) nestlings subjected to food shortage or food surplus under laboratory conditions. We assigned nestlings between 3 and 7 d of age to one of the following treatments: (1) food restriction, which resulted in a slowed growth at the rate found in undernourished, wild nestlings; (2) overfeeding, which totally suppressed begging; and (3) intermediate feeding. Threefold differences in energy consumption caused fivefold differences in body mass increments of the nestlings. Despite this, body mass-corrected RMR and intestinal mass were not affected by the feeding regime. The energy content of fecal output was highest in food-restricted birds, while their carcass fat content was lowest among treatment groups. Intestinal uptake rates of l-proline were low in the overfed and intermediate-fed young but significantly increased in the food-restricted birds, who attempted to maximize their rates of growth and development within the restrictive limits set by feeding regime. We noted a marked decrease of intestinal villi height in overfed birds as compared to intermediate-fed and food restricted nestlings. We conclude that song thrush nestlings are characterized by a limited plasticity of their developmental program, which prohibits overfed nestlings from significantly up-regulating their gut function to accommodate increased food intake. This suggests that they already grew at a rate close to their physiological maximum. We suggest two interpretations: (1) under natural conditions, song thrush nestlings do not face frequent, unpredictable fluctuations in food abundance that could select for developmental plasticity, or (2) strong selection for uniform adult phenotypes prevents flexible developmental trajectories, which would result in a diversity of adult phenotypes. PMID- 11009396 TI - Comparative and behavioral analyses of preferred speed: Anolis lizards as a model system. AB - I quantified the movement patterns of eight morphologically and ecologically diverse Caribbean Anolis lizard species in the field to address the following questions: (1) Do these eight species move at preferred speeds, and if so, what are these speeds? (2) What proportion of their maximum sprinting capacities do the anole species use when moving undisturbed? (3) What percentage of the time do lizards spend moving, and how far do they typically travel on a daily basis? (4) Have the preferred speeds of anoles coevolved with structural habitat use? Most of the distributions of speeds were highly skewed, with a preponderance of slow speed locomotion (<20% of maximum capacity). Median speeds varied almost eightfold among species, from a low of 4.9 cm/s (3.0% of maximum) to a high of 38.0 cm/s (22.4% of maximum). For all eight species, at least 75% of their locomotor movements took place between 0% and 40% of maximum capacity. The eight species varied almost 15-fold in the percentage of time they spent moving, indicating that not all anole species are equally sedentary. Through usage of modern comparative methods, I showed that Anolis species that move slowly through their environments also tend to use narrow perch diameters and have large habitat breadths. These findings show how evolutionary approaches can be profitably integrated with physiological data to understand how species use their habitats. PMID- 11009397 TI - Impact of externally attached loggers on the diving behaviour of the king penguin. AB - The impact of relatively small externally attached time series recorders on some foraging parameters of seabirds was investigated during the austral summer of 1995 by monitoring the diving behaviour of 10 free-ranging king penguins (Aptenodytes patagonicus) over one foraging trip. Time-depth recorders were implanted in the abdominal cavities of the birds, and half of the animals also had dummy loggers attached on their backs. Although most of the diving behaviour was not significantly affected by the external loggers (P>0.05), the birds with externally attached loggers performed almost twice as many shallow dives, between 0 and 10 m depth, as the birds without external loggers. These shallow dives interrupted more frequently the deep-diving sequences in the case of birds with external loggers (percentage of deep dives followed by deep dives: 46% for birds with implants only vs. 26% for birds with an external attachment). Finally, the distribution pattern of the postdive durations plotted against the hour of the day was more heterogeneous for the birds with an external package. In addition, these penguins had extended surfacing times between two deep dives compared to birds without external attachments (P<0.0001). These results suggest the existence of an extra energy cost induced by externally attached loggers. PMID- 11009398 TI - Time-course changes in the expression of Na+, K+-ATPase in gills and pyloric caeca of brown trout (Salmo trutta) during acclimation to seawater. AB - Changes in protein and mRNA expression of Na(+),K(+)-ATPase in gills and pyloric caeca of brown trout were investigated on a detailed time course after transfer from freshwater to 25 ppt seawater (SW). A transient deflection in plasma osmolality and muscle water content lasting from 4 h until day 3 was followed by restoration of hydromineral balance from day 5 onward. Gills and pyloric caeca responded to SW transfer by increasing Na(+),K(+)-ATPase activity from days 5 and 3, respectively, onward. In both tissues, this response was preceded by an increase in alpha-subunit Na(+), K(+)-ATPase mRNA as early as 12 h posttransfer. The similarity of the response in these two organs suggests that they both play significant physiological roles in restoring hydromineral balance after abrupt increase in salinity. Further, SW transfer induced a slight, though significant, increase in primary gill filament Na(+), K(+)-ATPase immunoreactive (NKIR) cell abundance. This was paralleled by a marked (50%) decrease in secondary lamellar NKIR cell abundance after less than 1 d in SW. Thus, SW acclimation in brown trout is characterised by a lasting decrease in overall NKIR cell abundance in the gill. We propose that SW transfer stimulates Na(+),K(+)-ATPase enzymatic activity within individual chloride cells long before (<1 d) it becomes apparent in measurements of whole-gill homogenate enzymatic activity. This is supported by the early stabilisation (12 h) of hydromineral balance. PMID- 11009399 TI - Intestinal nutrient uptake measurements and tissue damage: validating the everted sleeves method. AB - The reliability of methods for nutrient uptake measurements across the intestinal epithelium relies on the integrity of the mucosal epithelium and the enterocytes. We tested effects of tissue handling during the "everted sleeves method" on the length of intestinal villi, the surface magnification, the circumference of the gut, and the thickness of the muscle layer in sunbirds (Nectarinia osea), chicken (Gallus gallus), and mice (Mus domesticus). The sunbird has thin and delicate intestinal villi that are greatly affected by the everted sleeves method. After eversion and incubation, villi lost 30% of their original length. The severe tissue damage coincides with uptake measurements for glucose that were an order of magnitude lower than in other nectar-feeding (nectarivorous) birds of similar body size. Tissue handling during the everted sleeves method had significant effects on morphometric parameters of chicken and mouse intestines, but on a light-microscopical level, the tissue integrity and the cytology of the enterocytes were not altered. Therefore, we think that the everted sleeves method renders reliable and reproducible measurements of nutrient uptake in those species. We conclude that a histological evaluation is necessary to assess the reliability of the method before it is applied to adults or to the developmental stage of any species. PMID- 11009400 TI - The adjustment of avian metabolic rates and water fluxes to desert environments. AB - We tested the hypothesis that birds in arid environments, where primary productivity is low and surface water is scarce, have reduced energy expenditure and water loss compared with their mesic counterparts. Using both conventional least squares regression and regression based on phylogenetically independent contrasts, we showed that birds from desert habitats have reduced basal and field metabolic rates compared with species from mesic areas. Previous work showed that desert birds have reduced rates of total evaporative water loss when exposed to moderate environmental temperatures in the laboratory. We tested whether reduced rates of total evaporative water loss translate into low field water fluxes. Conventional ANCOVA indicated that desert birds have reduced water fluxes, but an analysis based on phylogenetically independent contrasts did not support this finding, despite the wide array of taxonomic affiliations of species in the data set. We conclude that the high ambient temperatures, the low primary productivity, and the water scarcity in desert environments have selected for or resulted in reduced rates of energy expenditure and evaporative water loss in birds that live in these climes. PMID- 11009401 TI - Xantusiid lizards have low energy, water, and food requirements. AB - Lizards in the family Xantusiidae (the night lizards) are known to have resting metabolic rates that are only half those of other lizards of comparable size. We evaluated whether xantusiids also have low field metabolic rates (FMR) and food requirements by measuring FMR and water flux rates with doubly labeled water in three xantusiid species in their natural habitats. Free-living Xantusia vigilis, Xantusia henshawi, and Xantusia riversiana processed energy and water very slowly, about one-third as fast as do other reptiles of similar size. Xantusiid lizards have a distinctive life history that is characterized by very slow growth and low reproductive rates, and they are intensely reclusive. This general lifestyle is also found in some species that live in environments with scarce food resources, such as in caves and in arid habitats, and these species may also have relatively low energy requirements. PMID- 11009402 TI - The effect of changing the gaseous diffusion coefficient on the mass loss pattern of Hyalophora cecropia pupae. AB - The importance of gas phase diffusion in insect gas exchange remains unclear. The role of diffusion in gas exchange of developing Hyalophora cecropia pupae was examined by altering the gaseous diffusion coefficient in the breathing mixture. Gaseous diffusion coefficients were manipulated by substituting helium or sulfur hexafluoride for the nitrogen usually present in air. Sensitive mass loss recordings were employed to monitor gas exchange activity. Mass loss recordings showed a two-phase cycle, open and closed-flutter. Mass loss rates during the open and closed-flutter periods were not altered in proportion to the changes induced in the rate of diffusion. Open-phase duration was inversely and proportionally related to the diffusion coefficient. These results are consistent with changes in spiracle resistance or convective flow during the open period in response to a change in the diffusion coefficient. In addition, they indicate a significant gas phase diffusive resistance within the pupal tracheal system. This previously unreported gas phase resistance appears to be a major determinant of the duration of the open period and thus of overall water loss rates in these pupae. PMID- 11009403 TI - Similarity of vasorelaxant effects of natriuretic peptides in isolated blood vessels of salmonids. AB - Natriuretic peptides (NPs) have been implicated in cardiovascular regulation in rainbow trout (Oncorhyncus mykiss), and it has been observed that the vasorelaxant activity of distinct trout and human NPs is similar in isolated trout arteries. This study characterizes the response of a variety of vessels from rainbow trout and other salmonids to different NPs. The effects of heterologous (rat atrial and human atrial) and homologous (rainbow trout atrial and rainbow trout ventricular) NPs were examined in precontracted efferent branchial arteries from rainbow trout (O. mykiss, Kamloops strain), lake whitefish (Coregonus clupeaformis), and in rainbow trout celiacomesenteric arteries and anterior cardinal veins. The response to mammalian NPs was also examined in efferent branchial arteries from the steelhead (O. mykiss, Skamania strain), coho salmon (Oncorhyncus kisutch), brook trout (Salvelinus fontinalis), and brown trout (Salmo trutta). In general, there were relatively few differences that were species, peptide, or vessel specific. There was no difference in the sensitivity (concentration producing a half-maximal response EC(50)) or efficacy (percent relaxation, i.e., E(max)) of trout or whitefish efferent branchial arteries to any NP, except human NP, which was significantly less effective (greater EC(50) and lower E(max)) in whitefish arteries. There were no differences in E(max) of mammalian NPs in efferent branchial arteries from any species, and only coho and brook trout had significantly different EC(50)'s (coho, 1.0+/-0.2 nM; brook trout, 4. 2+/-0.6 nM; and other species, from 1.9 to 3.5 nM). Rainbow and coho anterior cardinal veins were less sensitive than arteries to mammalian NPs (EC(50)'s; 8.8+/-2.0, 2.0+/-0.1 vs. 3.0+/-0.9, 1.0+/-0. 2, respectively), whereas brown trout veins were more sensitive (1. 0+/-0.2, 3.5+/-1.3, respectively). Sodium nitroprusside (SNP), which activates soluble guanylate cyclase, was vasodilatory, albeit significantly less potent than all NPs, in efferent branchial arteries of all species. SNP was significantly more potent in trout than whitefish efferent branchial arteries, whereas it was equally efficacious in these vessels. These results demonstrate that multiple vessels from various salmonids are similarly responsive to the vasorelaxant effects of a variety of NPs and that the salmonid NP receptor has relatively little ability to discriminate between homologous and heterologous peptides. We conclude that the vascular NP receptor complex is highly conserved among salmonids. Further, salmonids utilize cyclic guanosine monophosphate (cGMP) elevations for reductions of vascular tonus by both particulate and soluble guanylate cyclase pathways. PMID- 11009404 TI - Influence of environmental oxygen on development and hatching of aquatic eggs of the australian frog, Crinia georgiana. AB - The effect of oxygen partial pressure (Po(2)) on development and hatching was investigated in aquatic embryos of the myobatrachid frog, Crinia georgiana, in the field and in the laboratory. Eggs from 29 field nests experienced widely variable Po(2) but similar temperatures. Mean Po(2) in different nests ranged between 2.9 and 19.3 kPa (grand mean 12.9 kPa), and mean temperature ranged between 11.9 degrees and 16.8 degrees C (grand mean 13.7 degrees C). There was no detectable effect of Po(2) or temperature on development rate or hatching time in the field, except in one nest at 2.9 kPa where the embryos died, presumably in association with hypoxia. Laboratory eggs were incubated at 15 degrees C at a range of Po(2) between 2 and 25 kPa. Between 5 and 25 kPa, there was almost no effect of Po(2) on development rate to stage 26, but the embryos hatched progressively earlier-at earlier stages and lower gut-free body mass-at lower Po(2). At 2 kPa, development was severely delayed, growth of the embryo slowed, and morphological anomalies appeared. A high tolerance to low Po(2) may be an adaptation to embryonic development in the potentially hypoxic, aquatic environment. PMID- 11009405 TI - Conflicts between courtship and thermoregulation: the thermal ecology of amorous male garter snakes (Thamnophis sirtalis parietalis, colubridae). AB - Thermoregulatory behavior is an important component of daily activities for many reptiles, especially for small heliothermic (sun-basking) species that inhabit cold climates. However, the relative costs and benefits of thermoregulation depend on numerous factors, such that reptiles may sometimes accord a low priority to precise control of body temperatures. We observed and radio tracked garter snakes (Thamnophis sirtalis parietalis) in central Manitoba during the mating season (spring). Previous studies on this species have documented precise behavioral regulation of body temperatures during summer. In contrast, the courting snakes that we studied in springtime spent little time in overt thermoregulatory behavior. Body temperatures were extremely variable (both in outdoor enclosures and in the field) despite abundant opportunities for more precise thermal control. These small elongate reptiles cool so quickly (relative to the time periods needed for effective courtship) that any benefit to higher body temperatures would be transitory at best. Experiments show that hotter males are no better at obtaining matings or at detecting predators. Thus, male garter snakes concentrate on courtship rather than on basking. In the face of conflicting priorities, reptiles may often forgo precise thermoregulation because its benefits are too low, and its costs too high, compared with alternative behaviors. PMID- 11009406 TI - The energy budget of captive Siberian hamsters, Phodopus sungorus, exposed to photoperiod changes: mass loss is caused by a voluntary decrease in food intake. AB - Unlike most nonhibernating small mammals, Siberian hamsters (Phodopus sungorus) undergo a pronounced mass loss, almost exclusively as white adipose tissues, in response to a switch from long- to short-photoperiod exposure. This mass loss can be caused by an increase in the rate of energy expenditure, a decrease in the rate of energy intake, or a decrease in assimilation efficiency. In order to determine how they relate to photoperiod-induced mass loss, we measured these energy budget components every 2 wk on 12 captive Siberian hamsters exposed to 8 wk of long photoperiod followed by 12 wk of short photoperiod. Body mass decreased shortly after short-photoperiod exposure. This was accompanied by an immediate decrease in the rate of energy intake and, after a 2-wk delay, by a decrease in the rate of energy expenditure. The overall cumulative decrease in energy intake (376 kJ) could account for the mass loss (249 kJ) observed during short-photoperiod exposure. Regression analyses indicated that only the rate of energy intake was significantly related to the rate of mass change. Therefore, we conclude that photoperiod-induced mass loss in Siberian hamsters is caused by a decrease in the rate of food intake, rather than by changes in energy expenditure or in assimilation efficiency. PMID- 11009407 TI - Mechanism of delayed ovulation in a vespertilionid bat, scotophilus heathi: role of gonadotropin, insulin, and insulin-like growth factor-1. AB - The aim of this study was to evaluate the effects of luteinizing hormone (LH), follicle-stimulating hormone (FSH), insulin, and insulin-like growth factor-1 (IGF-1) on ovarian androstenedione synthesis to understand the mechanism responsible for delayed ovulation in Scotophilus heathi. We found that LH stimulated a dose-dependent increase in androstenedione synthesis by the ovary in vitro. This study also showed a clear seasonal variation in the ability of the ovary to produce androstenedione in vitro in response to LH and FSH stimulation. In response to LH and FSH, maximum quantities of androstenedione were produced during recrudescence in November. The same doses of gonadotropins during the preovulatory period in February stimulated comparatively low androstenedione secretion by the ovary. On the basis of these data, we suggest that in S. heathi, ovarian responsiveness to LH and FSH peaks during recrudescence. This study also showed a seasonal variation in the effects of insulin and IGF-1 on ovarian androstenedione production in vitro. Peak ovarian responsiveness to insulin and IGF-1 was observed during quiescence in September. It is hypothesized that increased insulin/IGF-1 sensitivity during September may be responsible for increased responsiveness to LH. Increased LH release, if coincident with the period of enhanced ovarian responsiveness to LH, may result in the excessive androstenedione production responsible for delayed ovulation in S. heathi. PMID- 11009408 TI - The quiet revolution: biodiversity informatics and the internet. AB - The massive development of biodiversity-related information systems on the Internet has created much that appears exciting but chaotic, a diversity to match biodiversity itself. This richness and the arrays of new sources are counterbalanced by the maddening difficulty in knowing what is where, or of comparing like with like. But quietly, behind the first waves of exuberance, biologists and computer scientists have started to pull together in a rising tide of coherence and organization. The fledgling field of biodiversity informatics looks set to deliver major advances that could turn the Internet into a giant global biodiversity information system. PMID- 11009409 TI - Interoperability of biodiversity databases: biodiversity information on every desktop. AB - Data about biodiversity are either scattered in many databases or reside on paper or other media not amenable to interactive searching. The Global Biodiversity Information Facility (GBIF) is a framework for facilitating the digitization of biodiversity data and for making interoperable an as-yet-unknown number of biodiversity databases that are distributed around the globe. In concert with other existing efforts, GBIF will catalyze the completion of a Catalog of the Names of Known Organisms and will develop search engines to mine the vast quantities of biodiversity data. It will be an outstanding tool for scientists, natural resource managers, and policy-makers. PMID- 11009410 TI - Flashing superluminal components in the jet of the radio galaxy 3C120 AB - A 16-month sequence of radio images of the active galaxy 3C120 with the Very Long Baseline Array reveals a region in the relativistic jet where superluminal components flash on and off over time scales of months, while the polarization angle rotates. This can be explained by interaction between the jet and an interstellar cloud located about 8 parsecs from the center of the galaxy. The cloud, which rotates the polarization direction and possibly eclipses a section of the jet, represents a "missing link" between the ultradense broad-emission line clouds closer to the center and the lower density narrow-emission-line clouds seen on kiloparsec scales. PMID- 11009411 TI - Quantum hall ferromagnetism in a two-dimensional electron system AB - Experiments on a nearly spin degenerate two-dimensional electron system reveals unusual hysteretic and relaxational transport in the fractional quantum Hall effect regime. The transition between the spin-polarized (with fill fraction nu = 1/3) and spin-unpolarized (nu = 2/5) states is accompanied by a complicated series of hysteresis loops reminiscent of a classical ferromagnet. In correlation with the hysteresis, magnetoresistance can either grow or decay logarithmically in time with remarkable persistence and does not saturate. In contrast to the established models of relaxation, the relaxation rate exhibits an anomalous divergence as temperature is reduced. These results indicate the presence of novel two-dimensional ferromagnetism with a complicated magnetic domain dynamic. PMID- 11009412 TI - Imaging coherent electron flow from a quantum point contact AB - Scanning a charged tip above the two-dimensional electron gas inside a gallium arsenide/aluminum gallium arsenide nanostructure allows the coherent electron flow from the lowest quantized modes of a quantum point contact at liquid helium temperatures to be imaged. As the width of the quantum point contact is increased, its electrical conductance increases in quantized steps of 2 e(2)/h, where e is the electron charge and h is Planck's constant. The angular dependence of the electron flow on each step agrees with theory, and fringes separated by half the electron wavelength are observed. Placing the tip so that it interrupts the flow from particular modes of the quantum point contact causes a reduction in the conductance of those particular conduction channels below 2 e(2)/h without affecting other channels. PMID- 11009413 TI - Technique for enhanced rare earth separation AB - A process is demonstrated for the efficient separation of rare earth elements, using a combination of selective reduction and vacuum distillation of halides. The large differences in the redox chemistry of the rare earth elements and in the vapor pressures of rare earth di- and trihalides are exploited for separation. Experimental proof of concept is provided for the binary systems praseodymium-neodymium and neodymium-samarium. This process enhances the separation factor for the isolation of samarium and neodymium from their mixture by more than an order of magnitude. PMID- 11009414 TI - A mechanical model for intraplate earthquakes: application to the new madrid seismic zone AB - We present a time-dependent model for the generation of repeated intraplate earthquakes that incorporates a weak lower crustal zone within an elastic lithosphere. Relaxation of this weak zone after tectonic perturbations transfers stress to the overlying crust, generating a sequence of earthquakes that continues until the zone fully relaxes. Simulations predict large (5 to 10 meters) slip events with recurrence intervals of 250 to 4000 years and cumulative offsets of about 100 meters, depending on material parameters and far-field stress magnitude. Most are consistent with earthquake magnitude, coseismic slip, recurrence intervals, cumulative offset, and surface deformation rates in the New Madrid Seismic Zone. Computed interseismic strain rates may not be detectable with available geodetic data, implying that low observed rates of strain accumulation cannot be used to rule out future damaging earthquakes. PMID- 11009415 TI - Dating of pore waters with (129)I: relevance for the origin of marine gas hydrates AB - Pore waters associated with gas hydrates at Blake Ridge in the Atlantic Ocean were dated by measuring their iodine-129/iodine ratios. Samples collected from sediments with ages between 1.8 and 6 million years ago consistently yield ages around 55 million years ago. These ages, together with the strong iodine enrichment observed in the pore waters, suggest that the origin of iodine is related to organic material of early Tertiary age, which probably is also the source of the methane in the gas hydrates at this location. PMID- 11009416 TI - Polyploidy and the evolution of gender dimorphism in plants. AB - Gender dimorphism and polyploidy are important evolutionary transitions that have evolved repeatedly in many plant families. We show that gender dimorphism in North American Lycium (Solanaceae) has evolved in polyploid, self-compatible taxa whose closest relatives are cosexual, self-incompatible diploids. This has occurred independently in South African Lycium. We present additional evidence for this pathway to gender dimorphism from 12 genera involving at least 20 independent evolutionary events. We propose that polyploidy is a trigger of unrecognized importance for the evolution of gender dimorphism, which operates by disrupting self-incompatibility and leading to inbreeding depression. Subsequently, male sterile mutants invade and increase because they are unable to inbreed. PMID- 11009417 TI - Alteration of stimulus-specific guard cell calcium oscillations and stomatal closing in Arabidopsis det3 mutant. AB - Cytosolic calcium oscillations control signaling in animal cells, whereas in plants their importance remains largely unknown. In wild-type Arabidopsis guard cells abscisic acid, oxidative stress, cold, and external calcium elicited cytosolic calcium oscillations of differing amplitudes and frequencies and induced stomatal closure. In guard cells of the V-ATPase mutant det3, external calcium and oxidative stress elicited prolonged calcium increases, which did not oscillate, and stomatal closure was abolished. Conversely, cold and abscisic acid elicited calcium oscillations in det3, and stomatal closure occurred normally. Moreover, in det3 guard cells, experimentally imposing external calcium-induced oscillations rescued stomatal closure. These data provide genetic evidence that stimulus-specific calcium oscillations are necessary for stomatal closure. PMID- 11009418 TI - High direct estimate of the mutation rate in the mitochondrial genome of Caenorhabditis elegans. AB - Mutations in the mitochondrial genome have been implicated in numerous human genetic disorders and offer important data for phylogenetic, forensic, and population genetic studies. Using a long-term series of Caenorhabditis elegans mutation accumulation lines, we performed a wide-scale screen for mutations in the mitochondrial genome that revealed a mutation rate that is two orders of magnitude higher than previous indirect estimates, a highly biased mutational spectrum, multiple mutations affecting coding function, as well as mutational hotspots at homopolymeric nucleotide stretches. PMID- 11009419 TI - Resetting of circadian time in peripheral tissues by glucocorticoid signaling. AB - In mammals, circadian oscillators reside not only in the suprachiasmatic nucleus of the brain, which harbors the central pacemaker, but also in most peripheral tissues. Here, we show that the glucocorticoid hormone analog dexamethasone induces circadian gene expression in cultured rat-1 fibroblasts and transiently changes the phase of circadian gene expression in liver, kidney, and heart. However, dexamethasone does not affect cyclic gene expression in neurons of the suprachiasmatic nucleus. This enabled us to establish an apparent phase-shift response curve specifically for peripheral clocks in intact animals. In contrast to the central clock, circadian oscillators in peripheral tissues appear to remain responsive to phase resetting throughout the day. PMID- 11009420 TI - Dendritic computation of direction selectivity by retinal ganglion cells. AB - Direction-selective ganglion cells (DSGCs) in the retina respond strongly when stimulated by image motion in a preferred direction but are only weakly excited by image motion in the opposite null direction. Such coding represents an early manifestation of complex information processing in the visual system, but the cellular locus and the synaptic mechanisms have yet to be elucidated. We recorded the synaptic activity of DSGCs using strategies to observe the asymmetric inhibitory inputs that underlie the generation of direction selectivity. The critical nonlinear interactions between the excitatory and inhibitory inputs took place postsynaptically within the dendrites of the DSGCs. PMID- 11009421 TI - Failure to regulate TNF-induced NF-kappaB and cell death responses in A20 deficient mice. AB - A20 is a cytoplasmic zinc finger protein that inhibits nuclear factor kappaB (NF kappaB) activity and tumor necrosis factor (TNF)-mediated programmed cell death (PCD). TNF dramatically increases A20 messenger RNA expression in all tissues. Mice deficient for A20 develop severe inflammation and cachexia, are hypersensitive to both lipopolysaccharide and TNF, and die prematurely. A20 deficient cells fail to terminate TNF-induced NF-kappaB responses. These cells are also more susceptible than control cells to undergo TNF-mediated PCD. Thus, A20 is critical for limiting inflammation by terminating TNF-induced NF-kappaB responses in vivo. PMID- 11009422 TI - A specificity-enhancing factor for the ClpXP degradation machine. AB - Events that stall bacterial protein synthesis activate the ssrA-tagging machinery, resulting in resumption of translation and addition of an 11-residue peptide to the carboxyl terminus of the nascent chain. This ssrA-encoded peptide tag marks the incomplete protein for degradation by the energy-dependent ClpXP protease. Here, a ribosome-associated protein, SspB, was found to bind specifically to ssrA-tagged proteins and to enhance recognition of these proteins by ClpXP. Cells with an sspB mutation are defective in degrading ssrA-tagged proteins, demonstrating that SspB is a specificity-enhancing factor for ClpXP that controls substrate choice. PMID- 11009423 TI - Ubiquitin-activating/conjugating activity of TAFII250, a mediator of activation of gene expression in Drosophila. AB - Ubiquitination of histones has been linked to the complex processes that regulate the activation of eukaryotic transcription. However, the cellular factors that interpose this histone modification during the processes of transcriptional activation are not well characterized. A biochemical approach identified the Drosophila coactivator TAFII250, the central subunit within the general transcription factor TFIID, as a histone-specific ubiquitin activating/conjugating enzyme (ubac). TAFII250 mediates monoubiquitination of histone H1 in vitro. Point mutations within the putative ubac domain of TAFII250 abolished H1-specific ubiquitination in vitro. In the Drosophila embryo, inactivation of the TAFII250 ubac activity reduces the cellular level of monoubiquitinated histone H1 and the expression of genes targeted by the maternal activator Dorsal. Thus, coactivator-mediated ubiquitination of proteins within the transactivation pathway may contribute to the processes directing activation of eukaryotic transcription. PMID- 11009425 TI - NF-kappaB-induced loss of MyoD messenger RNA: possible role in muscle decay and cachexia. AB - MyoD regulates skeletal muscle differentiation (SMD) and is essential for repair of damaged tissue. The transcription factor nuclear factor kappa B (NF-kappaB) is activated by the cytokine tumor necrosis factor (TNF), a mediator of skeletal muscle wasting in cachexia. Here, the role of NF-kappaB in cytokine-induced muscle degeneration was explored. In differentiating C2C12 myocytes, TNF-induced activation of NF-kappaB inhibited SMD by suppressing MyoD mRNA at the posttranscriptional level. In contrast, in differentiated myotubes, TNF plus interferon-gamma (IFN-gamma) signaling was required for NF-kappaB-dependent down regulation of MyoD and dysfunction of skeletal myofibers. MyoD mRNA was also down regulated by TNF and IFN-gamma expression in mouse muscle in vivo. These data elucidate a possible mechanism that may underlie the skeletal muscle decay in cachexia. PMID- 11009424 TI - Active remodeling of somatic nuclei in egg cytoplasm by the nucleosomal ATPase ISWI. AB - Cloning by the transplantation of somatic nuclei into unfertilized eggs requires a dramatic remodeling of chromosomal architecture. Many proteins are specifically lost from nuclei, and others are taken up from the egg cytoplasm. Recreating this exchange in vitro, we identified the chromatin-remodeling nucleosomal adenosine triphosphatase (ATPase) ISWI as a key molecule in this process. ISWI actively erases the TATA binding protein from association with the nuclear matrix. Defining the biochemistry of global nuclear remodeling may facilitate the efficiency of cloning and other dedifferentiation events that establish new stem cell lineages. PMID- 11009426 TI - Increase of maximum life-span in Sweden, 1861-1999. AB - A fundamental question in aging research is whether humans and other species possess an immutable life-span limit. We examined the maximum age at death in Sweden, which rose from about 101 years during the 1860s to about 108 years during the 1990s. The pace of increase was 0.44 years per decade before 1969 but accelerated to 1. 11 years per decade after that date. More than 70 percent of the rise in the maximum age at death from 1861 to 1999 is attributable to reductions in death rates above age 70. The rest are due to increased numbers of survivors to old age (both larger birth cohorts and increased survivorship from infancy to age 70). The more rapid rise in the maximum age since 1969 is due to the faster pace of old-age mortality decline during recent decades. PMID- 11009427 TI - Effects of chronic exercise on calcium signaling in rat vascular endothelium. AB - Chronic exercise enhances endothelium-dependent vasodilating responses. To investigate whether this is due to a change in endothelial Ca(2+) signaling, we examined intracellular Ca(2+) concentration ([Ca(2+)](i)) level in rat aortic endothelium in response to acetylcholine (ACh) or ATP. Four-week-old male Wistar rats were divided into control and exercise groups. The exercised animals ran on a treadmill at a moderate intensity for 60 min/day, 5 day/wk, for 10 wk. Rat aortas were then excised and loaded with fura 2. After the aortas were mounted on a flow chamber, these specimens were observed under an epifluorescence microscope equipped with ratio-imaging capability. Our results showed that 1) chronic exercise increased both ACh- and ATP-induced [Ca(2+)](i) responses; 2) ACh induced heterogeneous [Ca(2+)](i) elevation in individual endothelial cells; and 3) the exercise effect on ACh-evoked endothelial [Ca(2+)](i) elevation was inhibited by the Ca(2+) influx blocker SKF-96365, by a Ca(2+)-free buffer, or by high concentrations of extracellular K(+). We conclude that chronic exercise increases ACh-induced [Ca(2+)](i) elevation in rat aortic endothelium in situ, possibly by facilitating Ca(2+) influx. PMID- 11009428 TI - Aldose reductase inhibition alone or combined with an adenosine A(3) agonist reduces ischemic myocardial injury. AB - This study investigated whether aldose reductase (AR) inhibition with zopolrestat, either alone or in combination with an adenosine A(3)-receptor agonist (CB-MECA), reduced myocardial ischemic injury in rabbit hearts subjected to 30 min of regional ischemia and 120 min of reperfusion. Zopolrestat reduced infarct size by up to 61%, both in vitro (2 nM to 1 microM; EC(50) = 24 nM) and in vivo (50 mg/kg). Zopolrestat reduced myocardial sorbitol concentration (index of AR activity) by >50% (control, 15.0 +/- 2.2 nmol/g; 200 nM zopolrestat, 6.7 +/ 1.3 nmol/g). A modestly cardioprotective concentration of CB-MECA (0.2 nM) allowed a 50-fold reduction in zopolrestat concentration while providing a similar reduction in infarct size (infarct area/area at risk: control, 62 +/- 2%; 1 microM zopolrestat, 24 +/- 5%; 20 nM zopolrestat plus 0.2 nM CB-MECA, 20 +/- 4%). In conclusion, AR inhibition is cardioprotective both in vitro and in vivo. Furthermore, combining zopolrestat with an A(3) agonist allows a reduction in the zopolrestat concentration while maintaining an equivalent degree of cardioprotection. PMID- 11009429 TI - C-peptide exerts cardioprotective effects in myocardial ischemia-reperfusion. AB - Ischemia followed by reperfusion in the presence of polymorphonuclear leukocytes (PMNs) results in cardiac dysfunction. C-peptide, a cleavage product of proinsulin to insulin processing, induces nitric oxide (NO)-mediated vasodilation. NO is reported to attenuate cardiac dysfunction caused by PMNs after ischemia-reperfusion (I/R). Therefore, we hypothesized that C-peptide could attenuate PMN-induced cardiac dysfunction. We examined the effects of C-peptide in isolated ischemic (20 min) and reperfused (45 min) rat hearts perfused with PMNs. C-peptide (70 nmol/kg iv) given 4 or 24 h before I/R significantly improved coronary flow (P < 0.05), left ventricular developed pressure (LVDP) (P < 0.01), and the maximal rate of development of LVDP (+dP/dt(max)) compared with I/R hearts obtained from rats given 0.9% NaCl (P < 0.01). N(G)-nitro-L-arginine methyl ester (L-NAME) (50 micromol/l) blocked these cardioprotective effects. In addition, C-peptide significantly reduced cardiac PMN infiltration from 183 +/- 24 PMNs/mm(2) in untreated hearts to 44 +/- 10 and 58 +/- 25 PMNs/mm(2) in hearts from 4- and 24-h C-peptide-treated rats, respectively. Rat PMN adherence to rat superior mesenteric artery exposed to 2 U/ml thrombin was significantly reduced in rats given C-peptide compared with rats given 0.9% NaCl (P < 0.001). Moreover, C-peptide enhanced basal NO release from rat aortic segments. These results provide evidence that C-peptide can significantly attenuate PMN-induced cardiac contractile dysfunction in the isolated perfused rat heart subjected to I/R at least in part via enhanced NO release. PMID- 11009430 TI - Influence of erythrocyte aggregation on leukocyte margination in postcapillary venules of rat mesentery. AB - The role of erythrocyte (red blood cell; RBC) aggregation in affecting leukocyte (white blood cell; WBC) margination in postcapillary venules of the mesentery (rat) was explored by direct intravital microscopy. Optical techniques were refined and applied to relate the light-scattering properties of RBCs to obtain a quantitative index of aggregate size (G), which, under idealized conditions, represents the number of RBCs per aggregate. WBC margination, defined as the radial migration of WBCs to the venular wall and their subsequent rolling along the endothelium, was measured as the percentage of the potentially maximal WBC volumetric flux within the microvessel lumen (F(WBC)(*)). In normal blood, F(WBC)(*) increased exponentially fourfold, and G increased from 1 to 1.15 as wall shear rates () were reduced from a steady-state value of approximately 600 to <100 s(-1) by proximal occlusion with a blunt microprobe. Enhancement of aggregation by infusion (iv) of dextran 500 (428 kDa), to attain a systemic concentration of 3 g/100 ml, resulted in a four- and sevenfold increase in G and F(WBC)(*), respectively, as was reduced below 100 s(-1). Inhibition of RBC aggregation by infusion of dextran 40 (37.5 kDa) caused F(WBC)(*) to fall to one half of its steady-state level for < 100 s(-1). Thus it appears that the well known increase of WBC margination with reductions in is strongly dependent on the occurrence of RBC aggregation. Increasing the extent of RBC aggregation during reductions in also increased the firm adhesion of WBCs to the endothelium because of an enhanced probability of contact between leukocytes and the postcapillary venular wall. PMID- 11009431 TI - Adenosine A(2a)-receptor activation increases contractility in isolated perfused hearts. AB - Adenosine A(2a)-receptor activation enhances shortening of isolated cardiomyocytes. In the present study the effect of A(2a)-receptor activation on the contractile performance of isolated rat hearts was investigated by recording left ventricular pressure (LVP) and the maximal rate of LVP development (+dP/dt(max)). With constant-pressure perfusion, adenosine caused concentration dependent increases in LVP and +dP/dt(max), with detectable increases of 4.1 and 4.8% at 10(-6) M and maximal increases of 12.0 and 11.1% at 10(-4) M, respectively. The contractile responses were prevented by the A(2a)-receptor antagonists chlorostyryl-caffeine and aminofuryltriazolotriazinyl aminoethylphenol (ZM-241385) but were not affected by the beta(1)-adrenergic antagonist atenolol. The adenosine A(1)-receptor antagonist dipropylcyclopentylxanthine and pertussis toxin potentiated the positive inotropic effects of adenosine. The A(2a)-receptor agonists ethylcarboxamidoadenosine and dimethoxyphenyl-methylphenylethyl-adenosine also enhanced contractility. With constant-flow perfusion, 10(-5) M adenosine increased LVP and +dP/dt(max) by 5.5 and 6.0%, respectively. In the presence of the coronary vasodilator hydralazine, adenosine increased LVP and +dP/dt(max) by 7.5 and 7.4%, respectively. Dipropylcyclopentylxanthine potentiated the adenosine contractile responses with constant-flow perfusion in the absence and presence of hydralazine. These increases in contractile performance were also antagonized by chlorostyryl-caffeine and ZM-241385. The results indicate that adenosine increases contractile performance via activation of A(2a) receptors in the intact heart independent of beta(1)-adrenergic receptor activation or changes in coronary flow. PMID- 11009432 TI - cADP ribose and [Ca(2+)](i) regulation in rat cardiac myocytes. AB - cADP ribose (cADPR)-induced intracellular Ca(2+) concentration ([Ca(2+)](i)) responses were assessed in acutely dissociated adult rat ventricular myocytes using real-time confocal microscopy. In quiescent single myocytes, injection of cADPR (0.1-10 microM) induced sustained, concentration-dependent [Ca(2+)](i) responses ranging from 50 to 500 nM, which were completely inhibited by 20 microM 8-amino-cADPR, a specific blocker of the cADPR receptor. In myocytes displaying spontaneous [Ca(2+)](i) waves, increasing concentrations of cADPR increased wave frequency up to approximately 250% of control. In electrically paced myocytes (0.5 Hz, 5-ms duration), cADPR increased the amplitude of [Ca(2+)](i) transients in a concentration-dependent fashion, up to 150% of control. Administration of 8 amino-cADPR inhibited both spontaneous waves as well as [Ca(2+)](i) responses to electrical stimulation, even in the absence of exogenous cADPR. However, subsequent [Ca(2+)](i) responses to 5 mM caffeine were only partially inhibited by 8-amino-cADPR. In contrast, even under conditions where ryanodine receptor (RyR) channels were blocked with ryanodine, high cADPR concentrations still induced an [Ca(2+)](i) response. These results indicate that in cardiac myocytes, cADPR induces Ca(2+) release from the sarcoplasmic reticulum through both RyR channels and via mechanisms independent of RyR channels. PMID- 11009433 TI - Improved energy homeostasis of the heart in the metabolic state of exercise. AB - We postulate that metabolic conditions that develop systemically during exercise (high blood lactate and high nonesterified fatty acids) are favorable for energy homeostasis of the heart during contractile stimulation. We used working rat hearts perfused at physiological workload and levels of the major energy substrates and compared the metabolic and contractile responses to an acute low to-high work transition under resting versus exercising systemic metabolic conditions (low vs. high lactate and nonesterified fatty acids in the perfusate). Glycogen preservation, resulting from better maintenance of high-energy phosphates, was a consequence of improved energy homeostasis with high fat and lactate. We explained the result by tighter coupling between workload and total beta-oxidation. Total fatty acid oxidation with high fat and lactate reflected increased availability of exogenous and endogenous fats for respiration, as evidenced by increased long-chain fatty acyl-CoA esters (LCFA-CoAs) and by an increased contribution of triglycerides to total beta-oxidation. Triglyceride turnover (synthesis and degradation) also appeared to increase. Elevated LCFA CoAs caused high total beta-oxidation despite increased malonyl-CoA. The resulting bottleneck at mitochondrial uptake of LCFA-CoAs stimulated triglyceride synthesis. Our results suggest the following. First, both malonyl-CoA and LCFA CoAs determine total fatty acid oxidation in heart. Second, concomitant stimulation of peripheral glycolysis and lipolysis should improve cardiac energy homeostasis during exercise. We speculate that high lactate contributes to the salutary effect by bypassing the glycolytic block imposed by fatty acids, acting as an anaplerotic substrate necessary for high tricarbocylic acid cycle flux from fatty acid-derived acetyl-CoA. PMID- 11009434 TI - Endothelial adenosine transporter characterization in perfused guinea pig hearts. AB - Adenosine (Ado), a smooth muscle vasodilator and modulator of cardiac function, is taken up by many cell types via a saturable transporter, blockable by dipyridamole. To quantitate the influences of endothelial cells in governing the blood-tissue exchange of Ado and its concentration in the interstitial fluid, one must define the permeability-surface area products (PS) for Ado via passive transport through interendothelial gaps [PS(g)(Ado)] and across the endothelial cell luminal membrane (PS(ecl)) in their normal in vivo setting. With the use of the multiple-indicator dilution (MID) technique in Krebs-Ringer perfused, isolated guinea pig hearts (preserving endothelial myocyte geometry) and by separating Ado metabolites by HPLC, we found permeability-surface area products for an extracellular solute, sucrose, via passive transport through interendothelial gaps [PS(g)(Suc)] to be 1.9 +/- 0.6 ml. g(-1). min(-1) (n = 16 MID curves in 4 hearts) and took PS(g)(Ado) to be 1. 2 times PS(g)(Suc). MID curves were obtained with background nontracer Ado concentrations up to 800 micrometer, partially saturating the transporter and reducing its effective PS(ecl) for Ado. The estimated maximum value for PS(ecl) in the absence of background adenosine was 1.1 +/- 0.1 ml. g(-1). min(-1) [maximum rate of transporter conformational change to move the substrate from one side of the membrane to the other (maximal velocity; V(max)) times surface area of 125 +/- 11 nmol. g(-1). min(-1)], and the Michaelis-Menten constant (K(m)) was 114 +/- 12 microM, where +/- indicates 95% confidence limits. Physiologically, only high Ado release with hypoxia or ischemia will partially saturate the transporter. PMID- 11009435 TI - Mechanisms of endothelial cell swelling from lactacidosis studied in vitro. AB - One of the early sequelae of ischemia is an increase of circulating lactic acid that occurs in response to anaerobic metabolism. The purpose of the present study was to investigate whether lactic acidosis can induce endothelial swelling in vitro under closely controlled extracellular conditions. Cell volume of suspended cultured bovine aortic endothelial cells was measured by use of an advanced Coulter technique employing the "pulse area analysis" signal-processing technique (CASY1). The isosmotic reduction of pH from 7.4 to 6.8 had no effect on cell volume. Lowering of pH to 6.6, 6.4, or 6.0, however, led to significant, pH dependent increases of cell volume. Swelling was more pronounced in bicarbonate buffered media than in HEPES buffer. Specific inhibition of Na(+)/H(+) exchange by ethylisopropylamiloride completely prevented swelling in HEPES-buffered media. Pretreatment with ouabain to partially depolarize the cells did not affect the degree of acidosis-induced swelling. In bicarbonate-buffered media, the inhibition of transmembrane HCO(3)(-) transport by DIDS reduced swelling to a level comparable with that seen in the absence of bicarbonate ions. Lactacidosis induced endothelial swelling, therefore, is a result of intracellular pH regulatory mechanisms, namely, Na(+)/H(+) exchange and bicarbonate-transporting carriers. PMID- 11009436 TI - Transarterial wall oxygen gradients at the deployment site of an intra-arterial stent in the rabbit. AB - Intimal hyperplasia, common at the deployment site of an intra-arterial stent, may be caused by artery wall hypoxia. The purpose of this study was to determine the effect of an intra-arterial stent on artery wall oxygen concentrations. Transarterial wall oxygen gradients were measured by microelectrode at stent deployment sites in New Zealand White rabbits. Thinned artery walls and decreased oxygen tensions were noted throughout the artery wall immediately following stent deployment with a return toward control values at 28 days. Angioplasty alone had no acute effect on artery wall oxygen concentrations. Larger stent deployment diameters yielded acutely lower artery wall oxygen tensions. Using a linear one dimensional model for the oxygen profile, we noted that stent deployment resulted in acute changes in oxygen consumption in the inner artery wall that rapidly returned to control values. Changes were noted without differences in blood pressure or arterial blood oxygen concentrations. Oxygen delivery to and consumption within the artery wall are altered by intra-arterial stent deployment. A role for artery wall hypoxia in artery wall pathology at the deployment site of an intra-arterial stent is supported by these findings. PMID- 11009437 TI - Hypoxic pulmonary vasoconstriction is modified by P-450 metabolites. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P-450 4A (CYP4A) metabolite of arachidonic acid (AA) in human and rabbit lung microsomes and is a dilator of isolated human pulmonary arteries (PA). However, little is known regarding the contribution of P-450 metabolites to pulmonary vascular tone. We examined 1) the effect of two mechanistically distinct omega- and omega1 hydroxylase inhibitors on perfusion pressures in isolated rabbit lungs ventilated with normoxic or hypoxic gases, 2) changes in rabbit PA ring tone elicited by 20 HETE or omega- and omega1-hydroxylase inhibitors, and 3) expression of CYP4A protein in lung tissue. A modest increase in perfusion pressure (55 +/- 11% above normoxic conditions) was observed in isolated perfused lungs during ventilation with hypoxic gas (FI(O(2)) = 0.05). Inhibitors of 20-HETE synthesis, 17 oxydecanoic acid (17-ODYA) or N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), increased baseline perfusion pressure above that of vehicle and amplified hypoxia-induced increases in perfusion pressures by 92 +/- 11% and 105 +/- 11% over baseline pressures, respectively. 20-HETE relaxed phenylephrine (PE) constricted PA rings. Treatment with 17-ODYA enhanced PE-induced contraction of PA rings, consistent with inhibition of a product that promotes arterial relaxation, whereas 6-(20-propargyloxyphenyl)hexanoic acid (PPOH), an epoxygenase inhibitor, blunted contraction to PE. Conversion of AA into 20-HETE was blocked by 17-ODYA, DDMS, and hypoxia. CYP4A immunospecific protein confirms expression of CYP4A in male rabbit lung tissue. Our data suggest that endogenously produced 20-HETE could modify rabbit pulmonary vascular tone, particularly under hypoxic conditions. PMID- 11009438 TI - Alterations in myocardial collagen content affect rat papillary muscle function. AB - We investigated the influence of myocardial collagen volume fraction (CVF, %) and hydroxyproline concentration (microg/mg) on rat papillary muscle function. Collagen excess was obtained in 10 rats with unilateral renal ischemia for 5 wk followed by 3-wk treatment with ramipril (20 mg. kg(-1). day(-1)) (RHTR rats; CVF = 3.83 +/- 0. 80, hydroxyproline = 3.79 +/- 0.50). Collagen degradation was induced by double infusion of oxidized glutathione (GSSG rats; CVF = 2.45 +/- 0.52, hydroxyproline = 2.85 +/- 0.18). Nine untreated rats were used as controls (CFV = 3.04 +/- 0.58, hydroxyproline = 3.21 +/- 0.30). Active stiffness (AS; g. cm(-2). %L(max)(-1)) and myocyte cross-sectional area (MA; micrometer(2)) were increased in the GSSG rats compared with controls [AS 5.86 vs. 3.96 (P < 0.05); MA 363 +/- 59 vs. 305 +/- 28 (P < 0.05)]. In GSSG and RHTR groups the passive tension-length curves were shifted downwards, indicating decreased passive stiffness, and upwards, indicating increased passive stiffness, respectively. Decreased collagen content induced by GSSG is related to myocyte hypertrophy, decreased passive stiffness, and increased AS, and increased collagen concentration causes myocardial diastolic dysfunction with no effect on systolic function. PMID- 11009439 TI - Matrix metalloproteinase activity is required for activity-induced angiogenesis in rat skeletal muscle. AB - Proteolysis of the capillary basement membrane is a hallmark of inflammation mediated angiogenesis, but it is undetermined whether proteolysis plays a critical role in the process of activity-induced angiogenesis. Matrix metalloproteinases (MMPs) constitute the major class of proteases responsible for degradation of basement membrane proteins. We observed significant elevations of mRNA and protein levels of both MMP-2 and membrane type 1 (MT1)-MMP (2.9 +/- 0.7- and 1.5 +/- 0.1-fold above control, respectively) after 3 days of chronic electrical stimulation of rat skeletal muscle. Inhibition of MMP activity via the inhibitor GM-6001 prevented the growth of new capillaries as assessed by the capillary-to-fiber ratio (1.34 +/- 0.08 in GM-6001-treated muscles compared with 1.69 +/- 0.03 in control 7-day-stimulated muscles). This inhibition correlated with a significant reduction in the number of capillaries with observable breaks in the basement membrane, as assessed by electron microscopy (0.27 +/- 0.27% in GM-6001-treated muscles compared with 3.72 +/- 0.65% in control stimulated muscles). Proliferation of capillary-associated cells was significantly elevated by 2 days and remained elevated throughout 14 days of stimulation. Capillary associated cell proliferation during muscle stimulation was not affected by MMP inhibition (80.3 +/- 9.3 nuclei in control and 63.5 +/- 8.5 nuclei in GM-6001 treated animals). We conclude that MMP proteolysis of capillary basement membrane proteins is a critical component of physiological angiogenesis, and we postulate that capillary-associated proliferation precedes and occurs independently of endothelial cell sprout formation. PMID- 11009440 TI - Orthostatic hypotension in aging humans. AB - We tested the hypothesis that hypotension occurred in older adults at the onset of orthostatic challenge as a result of vagal dysfunction. Responses of heart rate (HR) and mean arterial pressure (MAP) were compared between 10 healthy older and younger adults during onset and sustained lower body negative pressure (LBNP). A younger group was also assessed after blockade of the parasympathetic nervous system with the use of atropine or glycopyrrolate and after blockade of the beta(1)-adrenoceptor by use of metoprolol. Baseline HR (older vs. younger: 59 +/- 4 vs. 54 +/- 1 beats/min) and MAP (83 +/- 2 vs. 89 +/- 3 mmHg) were not significantly different between the groups. During -40 Torr, significant tachycardia occurred at the first HR response in the younger subjects without hypotension, whereas significant hypotension [change in MAP (DeltaMAP) -7 +/- 2 mmHg] was observed in the elderly without tachycardia. After the parasympathetic blockade, tachycardiac responses of younger subjects were diminished and associated with a significant hypotension at the onset of LBNP. However, MAP was not affected after the cardiac sympathetic blockade. We concluded that the elderly experienced orthostatic hypotension at the onset of orthostatic challenge because of a diminished HR response. However, an augmented vasoconstriction helped with the maintenance of their blood pressure during sustained LBNP. PMID- 11009441 TI - IL-10 deficiency increases superoxide and endothelial dysfunction during inflammation. AB - Little is known about the role of interleukin-10 (IL-10), an anti-inflammatory cytokine, in blood vessels. We used IL-10-deficient mice (IL-10 -/-) to examine the hypothesis that IL-10 protects endothelial function after lipopolysaccharide (LPS) treatment. The responses of carotid arteries were studied in vitro 6 h after injection of a relatively low dose of LPS (10 microgram ip). In IL-10 -/- mice, the maximum relaxation to ACh (3 microM) was 56 +/- 6% (means +/- SE) after LPS injection and 84 +/- 4% after vehicle injection (P < 0.05). Thus endothelium dependent relaxation was impaired in carotid arteries from IL-10 -/- mice after LPS injection. In contrast, this dose of LPS did not alter relaxation to ACh in vessels from wild-type (IL-10 +/+) mice. Relaxation to nitroprusside and papaverine was similar in arteries from both IL-10 -/- and IL-10 +/+ mice after vehicle or LPS injection. Because inflammation is associated with increased levels of reactive oxygen species, we also tested the hypothesis that superoxide contributes to the impairment of endothelial function by LPS in the absence of IL 10. Results using confocal microscopy and hydroethidine indicated that levels of superoxide are elevated in carotid arteries from IL-10 -/- mice compared with IL 10 +/+ mice after LPS injection. The impaired relaxation of arteries from IL-10 /- mice after LPS injection was restored to normal by polyethylene glycol suspended superoxide dismutase (50 U/ml) or allopurinol (1 mM), an inhibitor of xanthine oxidase. These data provide direct evidence that IL-10 protects endothelial function after an acute inflammatory stimulus by limiting local increases in superoxide. The source of superoxide in this model may be xanthine oxidase. PMID- 11009442 TI - Na(+)/H(+) exchange inhibition-induced cardioprotection in dogs: effects on neutrophils versus cardiomyocytes. AB - Numerous studies have examined the effect of Na(+)/H(+) exchanger (NHE) inhibition on the myocardium; however, the effect of NHE-1 inhibition on neutrophil function has not been adequately examined. An in vivo canine model of myocardial ischemia-reperfusion injury in which 60 min of left anterior descending coronary artery occlusion followed by 3 h of reperfusion was used to examine the effect of NHE-1 inhibition on infarct size (IS) and neutrophil function. BIIB-513, a selective inhibitor of NHE-1, was infused before ischemia. IS was expressed as a percentage of area at risk (IS/AAR). NHE-1 inhibition significantly reduced IS/AAR and reduced neutrophil accumulation in the ischemic myocardium. NHE-1 inhibition attenuated both phorbol 12-myristate 13-acetate- and platelet-activating factor-induced neutrophil respiratory burst but not CD18 upregulation. Furthermore, NHE-1 inhibition directly protected cardiomyocytes against metabolic inhibition-induced lactate dehydrogenase release and hypercontracture. This study provides evidence that the cardioprotection induced by NHE-1 inhibition is likely due to specific protection of cardiomyocytes and attenuation of neutrophil activity. PMID- 11009443 TI - Three-week neonatal hypoxia reduces blood CGRP and causes persistent pulmonary hypertension in rats. AB - To increase understanding of persistent pulmonary hypertension, we examined chronic pulmonary effects of hypoxia at birth and their relationships with immunoreactive levels of the potent vasodilator, calcitonin gene-related peptide (CGRP). Rats were born in 10% hypobaric hypoxia, where they remained for 1-2 days, or in 15% hypoxia, where they remained for 21 days. All were then reared in normoxia for 3 mo followed by reexposure to 10% hypoxia for 7 days (H-->H) or continued normoxia (H-->N); age-matched normoxic rats were hypoxic for the last 7 days (N-->H) or normoxic throughout (N-->N). Results are as follows. Pulmonary arterial pressure (P(PA)) in 10% H-->N rats was normal at the end of the experiment (13 wk), but in rats reexposed to hypoxia (H-->H), pressure rose to 19% above N-->H controls. In 15% H-->N rats, P(PA) remained high, similar to that of N-->H rats, and increased further by 40% on reexposure (H-->H). Medial thickness of small pulmonary arteries in 10% H-->H rats also increased by 40% over N-->H controls and was equally high in 15% H-->N and H-->H rats. In N-->H rats from both experiments, right ventricular hypertrophy index (RVH) was increased after hypoxia at 15-16 wk. Also, in the 15% study, RVH remained elevated in H-->N rats and increased in H-->H rats by 19% above N-->H controls. Blood CGRP was reduced by neonate and adult hypoxia, and hypoxic reexposure (H- >H) further lowered blood CGRP in the 15% but not 10% study. Declining left ventricular blood CGRP correlated highly with logarithmically increasing P(PA) in the 15% study (r = -0.81, P = 0.000). In conclusion, 1) short perinatal exposure to 10% O(2) exacerbated pulmonary hypertension with hypoxia later in life, 2) 15% O(2) at birth and for 21 days caused persistent pulmonary hypertension and exacerbation with reexposure, and 3) P(PA) correlated highly with declining blood CGRP levels in the 15% study. PMID- 11009444 TI - Virtual sources associated with linear and curved strands of cardiac cells. AB - Transmembrane potential (V(m)) responses in cardiac strands with different curvature were characterized during uniform electric-field stimulation with the use of modeling and experimental approaches. Linear and U-shaped strands (width 100-150 micrometer) were stained with voltage-sensitive dye. V(m) was measured by optical mapping across the width and at sites of beginning curvature. Field pulses were applied transverse to the strands during the action-potential plateau. For linear strands, V(m) contained 1) a rapid passive component (V(m)(ar)) nearly linear and symmetric across the width, 2) a slower hyperpolarizing component (V(m)(as)) greater and faster on the anodal side, and 3) at high field strengths a delayed depolarizing component (V(m)(ad)) greater on the anodal side. For U-shaped strands, V(m) at sites of beginning curvature also contained rapid and slow components (V(m)(br) and V(m)(bs), respectively) that included contributions from the linear strand response and from the fiber curvature. V(m)(ar), V(m)(br), and part of V(m)(bs) could be attributed to passive behavior that was modeled, and V(m)(as), V(m)(ad), and part of V(m)(bs) could be attributed to active membrane currents. Thus curved strands exhibit field responses separable into components with characteristic amplitude, spatial, and temporal signatures. PMID- 11009445 TI - Skin microvascular adaptations during maturation and aging of hairless mice. AB - Using intravital fluorescence microscopy in the ears of hairless mice, we determined skin microvascular adaptations during the process of aging from juvenile to adult and senescent life (6-78 wk). Despite an increase of ear area within the first 36 wk, the number and branching pattern of both arteriolar and venular microvessels remained constant during the whole life period. Both arterioles and venules exhibited an increase in length, diameter, and intervascular distance up to the age of 36 wk. With the increase of the size of the ears, the observation that cutaneous capillary density remained unchanged implied new capillary formation. During aging to 78 wk, capillary density in the ears was reduced to approximately 40%. Functional analysis revealed an appropriate hyperemic response to a 2-min period of ischemia during late juvenile and adult life, which, however, was markedly reduced during senescence. Thus, except for capillaries, there is no indication for age-related new vessel formation. The process of aging from adult to senescent life does not cause any significant remodeling but is associated with a decrease of nutritive perfusion and a functional impairment to respond to stimuli such as ischemia. PMID- 11009446 TI - Effect of hypercholesterolemia on Ca(2+)-dependent K(+) channel-mediated vasodilatation in vivo. AB - Nitric oxide (NO)-mediated and NO-independent mechanisms of endothelium-dependent vasodilatation involve Ca(2+)-dependent K(+) (K(Ca)) channels. We examined the role in vivo of K(Ca) channels in NO-independent vasodilatation in hypercholesterolemia. Hindlimb vascular conductance was measured at rest and after aortic injection of ACh, bradykinin (BK), and sodium nitroprusside in anesthetized control and cholesterol-fed rabbits. Conductances were measured before and after treatment with the NO synthase antagonist N(omega)-nitro-L arginine methyl ester (L-NAME, 10 mg/kg) or K(Ca) blockers tetraethylammonium (30 mg/kg), charybdotoxin (10 microgram/kg), and apamin (50 microgram/kg). The contribution of NO to basal conductance was greater in control than in cholesterol-fed rabbits [2.2 +/- 0.4 vs. 1.1 +/- 0.3 (SE) ml. min(-1). kg(-1). 100 mmHg(-1), P < 0.05], but the NO-independent K(Ca) channel-mediated component was greater in the cholesterol-fed than in the control group (1.1 + 0.4 vs. 0.3 +/- 0.1 ml. min(-1). kg(-1). 100 mmHg(-1), P < 0.05). Maximum conductance response to ACh and BK was less in cholesterol-fed than in control rabbits, and the difference persisted after L-NAME (ACh: 7.7 +/- 0.7 vs. 10.1 +/- 0.5 ml. min( 1). kg(-1). 100 mmHg(-1), P < 0.005). Blockade of K(Ca) channels with tetraethylammonium or charybdotoxin + apamin almost completely abolished L-NAME resistant vasodilatation after ACh or BK. The magnitude of K(Ca)-mediated vasodilatation after ACh or BK was impaired in hypercholesterolemic rabbits. Vasodilator responses to nitroprusside did not differ between groups. In vivo, hypercholesterolemia is associated with an altered balance between NO-mediated and NO-independent K(Ca) channel contributions to resting vasomotor tone and impairment of both mechanisms of endothelium-dependent vasodilatation. PMID- 11009447 TI - K(ATP) channel activation reduces the severity of postresuscitation myocardial dysfunction. AB - Postresuscitation myocardial dysfunction has been recognized as a leading cause of the high postresuscitation mortality rate. We investigated the effects of ischemic preconditioning and activation of ATP-sensitive K(+) (K(ATP)) channels on postresuscitation myocardial function. Ventricular fibrillation (VF) was induced in 25 Sprague-Dawley rats. Cardiopulmonary resuscitation (CPR), including mechanical ventilation and precordial compression, was initiated after 4 min of untreated VF. Defibrillation was attempted after 6 min of CPR. The animals were randomized to five groups treated with 1) ischemic preconditioning, 2) K(ATP) channel opener, 3) ischemic preconditioning with K(ATP) channel blocker administered 1 min after VF, 4) K(ATP) channel blocker administered 45 min before induction of ischemic preconditioning, and 5) placebo. Postresuscitation myocardial function, as measured by the rate of left ventricular pressure increase at 40 mmHg, the rate of left ventricular decline, cardiac index, and duration of survival, was significantly improved in both preconditioned and K(ATP) channel opener-treated animals. K(ATP) channel blocker administered 45 min before induction of ischemic preconditioning completely abolished the myocardial protective effects of preconditioning. We conclude that ischemic preconditioning significantly improved post-CPR myocardial function and survival. These results also provide evidence that the myocardial protective effects of ischemic preconditioning are mediated by K(ATP) channel activation. PMID- 11009448 TI - P-450 epoxygenase and NO synthase inhibitors reduce cerebral blood flow response to N-methyl-D-aspartate. AB - Epoxyeicosatrienoic acids are cerebral vasodilators produced in astrocytes by cytochrome P-450 epoxygenase activity. The P-450 inhibitor miconazole attenuates the increase in cerebral blood flow (CBF) elicited by glutamate. We evaluated whether epoxygenase activity is involved in the CBF response to activation of the N-methyl-D-aspartate (NMDA) receptor subtype by using two structurally distinct inhibitors, miconazole and N-methylsulfonyl-6-(2-propargyloxyphenyl) hexanamide (MS-PPOH), a selective epoxygenase substrate inhibitor. Drugs were delivered locally through microdialysis probes in striata of anesthetized rats. Local CBF was measured by hydrogen clearance and compared with CBF in contralateral striatum receiving vehicle. Microdialysis perfusion of NMDA doubled CBF and increased nitric oxide (NO) production estimated by recovery of labeled citrulline in the dialysate during labeled arginine infusion. Perfusion of miconazole or MS-PPOH blocked the increase in CBF without decreasing citrulline recovery. Perfusion of N(omega)-nitro-L-arginine decreased baseline CBF and inhibited the CBF response to NMDA. Perfusion of MS-PPOH did not inhibit the CBF response to sodium nitroprusside. We conclude that both the P-450 epoxygenase and NO synthase pathways are involved in the local CBF response to NMDA receptor activation, and that the signaling pathway may be more complex than simply NO diffusion from neurons to vascular smooth muscle. PMID- 11009449 TI - VEGF and ATP act by different mechanisms to increase microvascular permeability and endothelial [Ca(2+)](i). AB - Vascular endothelial growth factor (VEGF) increases hydraulic conductivity (L(p)) by stimulating Ca(2+) influx into endothelial cells. To determine whether VEGF mediated Ca(2+) influx is stimulated by release of Ca(2+) from intracellular stores, we measured the effect of Ca(2+) store depletion on VEGF-mediated increased L(p) and endothelial intracellular Ca(2+) concentration ([Ca(2+)](i)) of frog mesenteric microvessels. Inhibition of Ca(2+) influx by perfusion with NiCl(2) significantly attenuated VEGF-mediated increased [Ca(2+)](i). Depletion of Ca(2+) stores by perfusion of vessels with thapsigargin did not affect the VEGF-mediated increased [Ca(2+)](i) or the increase in L(p). In contrast, ATP mediated increases in both [Ca(2+)](i) and L(p) were inhibited by thapsigargin perfusion, demonstrating that ATP stimulated store-mediated Ca(2+) influx. VEGF also increased Mn(2+) influx after perfusion with thapsigargin, whereas ATP did not. These data showed that VEGF increased [Ca(2+)](i) and L(p) even when Ca(2+) stores were depleted and under conditions that prevented ATP-mediated increases in [Ca(2+)](i) and L(p). This suggests that VEGF acts through a Ca(2+) store independent mechanism, whereas ATP acts through Ca(2+) store-mediated Ca(2+) influx. PMID- 11009450 TI - Significance of ERK cascade compared with JAK/STAT and PI3-K pathway in gp130 mediated cardiac hypertrophy. AB - We compared the role of the Raf-1/mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MEK)/extracellular signal-regulated protein kinase (ERK)/p90(RSK) cascade in gp130-mediated cardiac hypertrophy with the contribution of the Janus kinase (JAK)/signal transduction and activation of transcription (STAT) and phosphatidylinositide 3-kinase (PI3-K) pathways. Primary cultured neonatal rat cardiomyocytes were stimulated with leukemia inhibitory factor (LIF). LIF sequentially activated Raf-1, MEK1/2, ERK1/2, and p90(RSK). We used PD-98059 (a specific MEK inhibitor), AG-490 (a JAK2 inhibitor), and wortmannin (a PI3-K inhibitor) to confirm that this cascade was independent of the JAK/STAT and PI3-K/p70 S6 kinase (S6K) pathways. PD-98059, AG-490, and wortmannin suppressed the LIF-induced increase in [(3)H]phenylalanine uptake by 54.7, 21.5, and 25.6%, respectively, and inhibited the increase in cell area by 61.2, 42.8, and 39.2%, respectively. Reorganization of myofilaments was predominantly suppressed by AG-490. LIF-induced expression of c-fos, brain natriuretic peptide, and skeletal alpha-actin mRNA was markedly suppressed by PD 98059 and moderately suppressed by wortmannin and AG-490. Atrial natriuretic peptide was significantly suppressed by AG-490. These findings indicate that this pathway is critically involved in protein synthesis, induction of c-fos, brain natriuretic peptide, and skeletal alpha-actin expression and is partially involved in myofilament reorganization and atrial natriuretic peptide induction in gp130-mediated cardiac hypertrophy. PMID- 11009451 TI - Model of structural and functional adaptation of small conductance vessels to arterial hypotension. AB - Vascular networks adapt structurally in response to local pressure and flow and functionally in response to the changing needs of tissue. Whereas most research has either focused on adaptation of the macrocirculation, which primarily transports blood, or the microcirculation, which primarily controls flow, the present work addresses adaptation of the small conductance vessels in between, which both conduct blood and resist flow. A simple hemodynamic model is introduced consisting of three parts: 1) bifurcating arterial and venous trees, 2) an empirical description of the microvasculature, and 3) a target shear stress depending on pressure. This simple model has the minimum requirements to explain qualitatively the observed structure in normotensive conditions. It illustrates that flow regulation in the microvasculature makes adaptation in the larger conductance vessels stable. Furthermore, it suggests that structural changes in response to hypotension can account for the observed decrease in the lower limit of autoregulation in chronically hypotensive vasculature. Independent adaptation to local conditions thus yields a coordinated set of structural changes that ultimately adapts supply to demand. PMID- 11009452 TI - Mechanisms of calcitonin gene-related peptide-induced increases of pulmonary blood flow in fetal sheep. AB - Fetal pulmonary blood flow is regulated by various vasoactive substances. One, calcitonin gene-related peptide (CGRP), increases pulmonary blood flow. We examined four key physiological mechanisms underlying this response using the blocker drugs CGRP receptor blocker (CGRP(8-37)), nitric oxide synthase inhibitor [N(omega)-nitro-L-arginine (L-NNA)], adenosine triphosphate-dependent potassium (K(ATP)) channel blocker (glibenclamide), and cyclooxygenase inhibitor (indomethacin) in 17 near-term fetal sheep. Catheters were placed in the left (LPA) and main pulmonary arteries, and an ultrasonic flow transducer was placed around the LPA to measure flow continuously. CGRP was injected directly into the LPA (mean 1.02 microgram/kg) before and after blockade, and responses to CGRP were statistically compared. Before blockade, CGRP increased LPA blood flow from 23 +/- 25 to 145 +/- 77 ml/min (means +/- SD), and these increases were significantly attenuated by CGRP(8-37) (n = 6; 91% inhibition), L-NNA (n = 6; 86% inhibition), and glibenclamide (n = 6; 69% inhibition). No significant changes were found with indomethacin (n = 6; 4% inhibition). Thus, in the fetal pulmonary circulation, CGRP increases pulmonary blood flow not only through its specific receptor but also, in part, through nitric oxide release and K(ATP) channel activation. PMID- 11009453 TI - Role of protein kinase C in alpha(1)-adrenergic regulation of a(Na)(i) in guinea pig ventricular myocytes. AB - We investigated the role of protein kinase C (PKC) in alpha(1)-adrenergic regulation of intracellular Na(+) activity (a(Na)(i)) in single guinea pig ventricular myocytes. a(Na)(i) and membrane potentials were measured with the Na(+)-sensitive indicator sodium-binding benzofuran isophthalate and conventional microelectrodes, respectively, at room temperature (24-26 degrees C) while myocytes were stimulated at a rate of 0.25-0.3 Hz. The PKC activator 4beta phorbol 12-myristate 13-acetate (PMA) decreased a(Na)(i) in a concentration dependent manner. PMA (100 nM) produced a maximal decrease in a(Na)(i) of 1.5 mM from 6.5 +/- 0.4 to 5.0 +/- 0.4 mM (means +/- SE, n = 12, P < 0.01). The PMA concentration required for a half-maximal decrease in a(Na)(i) was 0.46 +/- 0.13 nM (n = 3, P < 0.01). An inactive phorbol, 4alpha-phorbol 12-myristate 13 acetate, did not decrease a(Na)(i). The decrease caused by PMA could be blocked by the PKC inhibitors staurosporine and bisindolylmaleimide I (GF-109203X). Stimulation of the alpha(1)-adrenoceptor with 50 microM phenylephrine decreased a(Na)(i) from 6.1 +/- 0.3 to 4.6 +/- 0.3 mM (n = 11, P < 0.01). The decrease in a(Na)(i) produced by phenylephrine was blocked by pretreatment with staurosporine, GF-109203X, or PMA. The decrease in a(Na)(i) produced by PMA was not prevented by pretreatment with tetrodotoxin but was blocked by pretreatment with strophanthidin or high extracellular K(+) concentration. The results suggest that alpha(1)-adrenergic receptor activation results in a decrease in a(Na)(i) via PKC-induced stimulation of the Na(+)-K(+) pump in cardiac myocytes. PMID- 11009454 TI - Nonlinear dynamics of heart rate variability during experimental hemorrhage in ketamine-anesthetized rats. AB - Indexes of heart rate variability (HRV) based on linear stochastic models are independent risk factors for arrhythmic death (AD). An index based on a nonlinear deterministic model, a reduction in the point correlation dimension (PD2i), has been shown in both animal and human studies to have a higher sensitivity and specificity for predicting AD. Dimensional reduction subsequent to transient ischemia was examined previously in a simple model system, the intrinsic nervous system of the isolated rabbit heart. The present study presents a new model system in which the higher cerebral centers are blocked chemically (ketamine inhibition of N-methyl-D-aspartate receptors) and the system is perturbed over a longer 15-min interval by continuous hemorrhage. The hypothesis tested was that dimensional reduction would again be evoked, but in association with a more complex relationship between the system variables. The hypothesis was supported, and we interpret the greater response complexity to result from the larger autonomic superstructure attached to the heart. The complexities observed in the nonlinear heartbeat dynamics constitute a new genre of autonomic response, one clearly distinct from a hardwired reflex or a cerebrally determined defensive reaction. PMID- 11009455 TI - PKCepsilon modulates NF-kappaB and AP-1 via mitogen-activated protein kinases in adult rabbit cardiomyocytes. AB - We have previously shown that protein kinase C (PKC)-epsilon, nuclear factor (NF) kappaB, and mitogen-activated protein kinases (MAPKs) are essential signaling elements in ischemic preconditioning. In the present study, we examined whether activation of PKCepsilon affects the activation of NF-kappaB in cardiac myocytes and whether MAPKs are mediators of this signaling event. Activation of PKCepsilon (+108% above control) in adult rabbit cardiomyocytes to a degree that has been previously shown to protect myocytes against hypoxic injury increased the DNA binding activity of NF-kappaB (+164%) and activator protein (AP)-1 (+127%) but not that of Elk-1. Activation of PKCeta did not have an effect on these transcription factors. Activation of PKCepsilon also enhanced the phosphorylation activities of the p44/p42 MAPKs and the p54/p46 c-Jun NH(2)-terminal kinases (JNKs). PKCepsilon-induced activation of NF-kappaB and AP-1 was completely abolished by inhibition of the p44/p42 MAPK pathway with PD98059 and by inhibition of the p54/p46 JNK pathway with a dominant negative mutant of MAPK kinase-4, indicating that both signaling pathways are necessary. Taken together, these data identify NF-kappaB and AP-1 as downstream targets of PKCepsilon, thereby establishing a molecular link between activation of PKCepsilon and activation of NF-kappaB and AP-1 in cardiomyocytes. The results further demonstrate that both the p44/p42 MAPK and the p54/p46 JNK signaling pathways are essential mediators of this event. PMID- 11009456 TI - Cardioprotection by K(ATP) channels in wild-type hearts and hearts overexpressing A(1)-adenosine receptors. AB - We studied the role of mitochondrial ATP-sensitive K(+) (K(ATP)) channels in modifying functional responses to 20 min global ischemia and 30 min reperfusion in wild-type mouse hearts and in hearts with approximately 250-fold overexpression of functionally coupled A(1)-adenosine receptors (A(1)ARs). In wild-type hearts, time to onset of contracture (TOC) was 303 +/- 24 s, with a peak contracture of 89 +/- 5 mmHg. Diastolic pressure remained elevated at 52 +/- 6 mmHg after reperfusion, and developed pressure recovered to 40 +/- 6% of preischemia. A(1)AR overexpression markedly prolonged TOC to 517 +/- 84 s, reduced contracture to 64 +/- 6 mmHg, and improved recovery of diastolic (to 9 +/ 4 mmHg) and developed pressure (to 82 +/- 8%). 5-Hydroxydecanoate (5-HD; 100 microM), a mitochondrial K(ATP) blocker, did not alter ischemic contracture in wild-type hearts, but increased diastolic pressure to 69 +/- 8 mmHg and reduced developed pressure to 10 +/- 5% during reperfusion. In transgenic hearts, 5-HD reduced TOC to 348 +/- 18 s, increased postischemic contracture to 53 +/- 4 mmHg, and reduced recovery of developed pressure to 22 +/- 4%. In summary, these data are the first to demonstrate that endogenous activation of K(ATP) channels improves tolerance to ischemia-reperfusion in murine myocardium. This functional protection occurs without modification of ischemic contracture. The data also support a role for mitochondrial K(ATP) channel activation in the pronounced cardioprotection afforded by overexpression of myocardial A(1)ARs. PMID- 11009457 TI - Validation of a mouse conductance system to determine LV volume: comparison to echocardiography and crystals. AB - The application of left ventricular pressure-volume analysis to transgenic mice to characterize the cardiac phenotype has been problematic due to the small size of the mouse heart and the rapid heartbeat. Conductance technology has been miniaturized for the mouse and can solve this problem. However, there has been no validation of this technique. Accordingly, we performed echocardiography followed by simultaneous ultrasonic crystals, flow probe, and conductance studies in 18 CD 1 mice. Raw conductance volumes were corrected for an inhomogenous electrical field (alpha) and parallel conductance (G(pi)) yielding a stroke volume of 14.1 +/- 3.7 microliter/beat, end-diastolic volume of 20.8 +/- 6.5 microliter, and end systolic volume of 9.0 +/- 5.8 microliter. The mean conductance volumes were no different from those derived by flow probe and echocardiography but did differ from ultrasonic crystals. G(pi) was determined to be 14.9 +/- 8.7 microliter. However, hypertonic saline altered dimension and pressure in the mouse left ventricle. Although G(pi) can be determined by the hypertonic saline method, saline altered hemodynamics, questioning its validity in the mouse. Although mean measures of absolute volume may be similar among different techniques, individual values did not correlate. PMID- 11009458 TI - Reduced contractile response to insulin and IGF-I in ventricular myocytes from genetically obese Zucker rats. AB - Obesity plays a pivotal role in the pathophysiology of metabolic and cardiovascular diseases. Resistance to insulin is commonly seen in metabolic disorders such as obesity and diabetes. Insulin-like growth factor-I (IGF-I) mimics insulin in many tissues and has been shown to enhance cardiac contractile function and growth. Because IGF-I resistance often accompanies resistance to insulin, we sought to determine whether IGF-I-induced myocardial contractile was elevated and whether heart and kidney size were enlarged in obese compared with lean rats. The myocyte contraction profile in the obese rats showed a decreased peak shortening associated with prolonged relengthening and normal shortening duration, a pattern similar to that observed in diabetes. IGF-I (1-500 ng/ml) caused a dose-dependent increase in peak shortening in lean but not obese animals, but it did not alter the duration of shortening and relengthening. Consistent with contractile data, IGF-I induced a dose-dependent increase in Ca(2+) transients only in myocytes of lean rats. IGF-I receptor mRNA levels were significantly reduced in obese rat hearts. These results suggest that the IGF-I induced cardiac contractile responses are attenuated in the Zucker model of obesity. The mechanisms underlying this alteration may be related to the decreased receptor number and/or changes in intracellular Ca(2+) handling in these animals. PMID- 11009459 TI - Time course of vasodilatory responses in skeletal muscle arterioles: role in hyperemia at onset of exercise. AB - At the onset of dynamic exercise, muscle blood flow increases within 1-2 s. It has been postulated that local vasodilatory agents produced by the vascular endothelium or the muscle itself contribute to this response. We hypothesized that only vasodilators that act directly on the vascular smooth muscle could produce vasodilation of skeletal muscle arterioles in <2 s. To test this hypothesis, we determined the time course of the vasodilatory response of isolated skeletal muscle arterioles to direct application of potassium chloride, adenosine, acetylcholine, and sodium nitroprusside. Soleus and gastrocnemius muscles were dissected from the hindlimbs of male Sprague-Dawley rats. First order arterioles (100-200 microm) were isolated, cannulated on micropipettes, and pressurized to 60 cmH(2)O in an organ bath. Vasodilatory agents were added directly to the bath, and diameter responses of the arterioles were recorded in real time on a videotape recorder. Frame-by-frame analysis of the diameter responses indicated that none of the vasodilator agents tested produced significant diameter increases in <4 s in either soleus or gastrocnemius muscle arterioles. These results indicate that, although these local vasodilators produce significant vasodilation of skeletal muscle resistance arterioles, these responses are not rapid enough (within 1-2 s) to contribute to the initiation of the exercise hyperemic response at the onset of dynamic exercise. PMID- 11009460 TI - Starling forces that oppose filtration after tissue oncotic pressure is increased. AB - We tested the hypothesis that the effective oncotic force that opposes fluid filtration across the microvessel wall is the local oncotic pressure difference across the endothelial surface glycocalyx and not the global difference between the plasma and tissue. In single frog mesenteric microvessels perfused and superfused with solutions containing 50 mg/ml albumin, the effective oncotic pressure exerted across the microvessel wall was not significantly different from that measured when the perfusate alone contained albumin at 50 mg/ml. Measurements were made during transient and steady-state filtration at capillary pressures between 10 and 35 cmH(2)O. A cellular-level model of coupled water and solute flows in the interendothelial cleft showed water flux through small breaks in the junctional strand limited back diffusion of albumin into the protected space on the tissue side of the glycocalyx. Thus oncotic forces opposing filtration are larger than those estimated from blood-to-tissue protein concentration differences, and transcapillary fluid flux is smaller than estimated from global differences in oncotic and hydrostatic pressures. PMID- 11009461 TI - Endocardial activation during ventricular fibrillation in normal and failing canine hearts. AB - Because congestive heart failure (CHF) promotes ventricular fibrillation (VF), we compared VF in seven dogs with CHF induced by combined myocardial infarction and rapid ventricular pacing to VF in six normal dogs. A noncontact, multielectrode array balloon catheter provided full-surface real-time left ventricular (LV) endocardial electrograms and a dynamic color-coded display of endocardial activation projected onto a three-dimensional model of the LV. Fast Fourier transform (FFT) analysis of virtual electrograms showed no difference in peak or centroid frequency in CHF dogs compared with normals. The average number of simultaneous noncontiguous wavefronts present during VF was higher in normals (2.4 +/- 1.0 at 10 s of VF) than in CHF dogs (1.3 +/- 1.0, P < 0.005) and decreased in both over time. The wavefront "turnover" rate, estimated using FFT of the noncontiguous wavefront data, did not differ between normals and CHF and did not change over 5 min of VF. Thus the fundamental frequency characteristics of VF are unaltered by CHF, but dilated abnormal ventricles sustain fewer active wavefronts than do normal ventricles. PMID- 11009462 TI - Novel characteristics of a misprocessed mutant HERG channel linked to hereditary long QT syndrome. AB - Hereditary long QT syndrome (hLQTS) is a heterogeneous genetic disease characterized by prolonged QT interval in the electrocardiogram, recurrent syncope, and sudden cardiac death. Mutations in the cardiac potassium channel HERG (KCNH2) are the second most common form of hLQTS and reduce the delayed rectifier K(+) currents, thereby prolonging repolarization. We studied a novel COOH-terminal missense mutation, HERG R752W, which segregated with the disease in a family of 101 genotyped individuals. When the mutant cRNA was expressed in Xenopus oocytes it produced enhanced rather than reduced currents. Simulations using the Luo-Rudy model predicted minimal shortening rather than prolongation of the cardiac action potential. Consequently, a normal or shortened QT interval would be expected in contrast to the long QT observed clinically. This anomaly was resolved by our observation that the mutant protein was not delivered to the plasma membrane of mammalian cells but was retained intracellularly. We found that this trafficking defect was corrected at lower incubation temperatures and that functional channels were now delivered to the plasma membrane. However, trafficking could not be restored by chemical chaperones or E-4031, a specific blocker of HERG channels. Therefore, HERG R752W represents a new class of trafficking mutants in hLQTS. The occurrence of different classes of misprocessed channels suggests that a unified therapeutic approach for altering HERG trafficking will not be possible and that different treatment modalities will have to be matched to the different classes of trafficking mutants. PMID- 11009463 TI - Upregulation of neuronal nitric oxide synthase in skeletal muscle by swim training. AB - Exercise enhances cardiac output and blood flow to working skeletal muscles but decreases visceral perfusion. The alterations in nitric oxide synthase (NOS) activity and/or expression of the cardiopulmonary, skeletal muscle, and visceral organs induced by swim training are unknown. In sedentary and swim-trained rats (60 min twice/day for 3-4 wk), we studied the alterations in NOS in different tissues along with hindquarter vasoreactivity in vivo during rest and mesenteric vascular bed reactivity in vitro. Hindquarter blood flow and conductance were reduced by norepinephrine in both groups to a similar degree, whereas N(G)-nitro L-arginine methyl ester reduced both indexes to a greater extent in swim-trained rats. Vasodilator responses to ACh, but not bradykinin or S-nitroso-N-acetyl penicillamine, were increased in swim-trained rats. Ca(2+)-dependent NOS activity was enhanced in the hindquarter skeletal muscle, lung, aorta, and atria of swim trained rats together with increased expression of neuronal NOS in the hindquarter skeletal muscle and endothelial NOS in the cardiopulmonary organs. Mesenteric arterial bed vasoreactivity was unaltered by swim training. Physiological adaptations to swim training are characterized by enhanced hindquarter ACh-induced vasodilation with upregulation of neuronal NOS in skeletal muscle and endothelial NOS in the lung, atria, and aorta. PMID- 11009464 TI - Mathematical analysis of canine atrial action potentials: rate, regional factors, and electrical remodeling. AB - Dogs have been used extensively to study atrial arrhythmias, but there are no published mathematical models of the canine atrial action potential (AP). To obtain insights into the ionic mechanisms governing canine atrial AP properties, we incorporated formulations of K(+), Na(+), Ca(2+), and Cl(-) currents, based on measurements in canine atrial myocytes, into a mathematical model of the AP. The rate-dependent behavior of model APs corresponded to experimental measurements and pointed to a central role for L-type Ca(2+) current inactivation in rate adaptation. Incorporating previously described regional ionic current variations into the model largely reproduced AP forms characteristic of the corresponding right atrial regions (appendage, pectinate muscle, crista terminalis, and atrioventricular ring). When ionic alterations induced by tachycardia-dependent remodeling were incorporated, the model reproduced qualitatively the AP features constituting the cellular substrate for atrial fibrillation. We conclude that this ionic model of the canine atrial AP agrees well with experimental measurements and gives potential insights into mechanisms underlying functionally important electrophysiological phenomena in canine atrium. PMID- 11009465 TI - Alterations of endothelium and smooth muscle function in monocrotaline-induced pulmonary hypertensive arteries. AB - We examined how monocrotaline (MCT), which impairs the endothelium and causes pulmonary hypertension, altered the endothelial regulation of pulmonary artery functions. Rats were given a single injection of MCT (60 mg/kg sc). Pulmonary arteries were depolarized to -48.3 +/- 2.6 and -39.8 +/- 2.2 mV at 2 and 3 wk after treatment with MCT, respectively (control arteries -59.9 +/- 1.9 mV). The basal tone in the resting state was only slightly elevated at 3 wk in endothelium intact arteries. Removal of the endothelium caused further depolarization in MCT affected arteries at 2 wk, but not at 3 wk, and greatly elevated the basal tone at 2 and 3 wk. N(omega)-nitro-L-arginine (200 microM), a nitric oxide synthase inhibitor, also caused depolarization in endothelium-intact arteries in both groups and elevated the basal tone of MCT-affected arteries. The relaxant responses of pulmonary arteries to ACh and A-23187 were depressed at 2 and 3 wk after MCT treatment. Thus chronic impairment of the endothelium altered the property of the pulmonary artery leading to depolarization. During the early stage of depolarization, a rise in the basal tone was offset by nitric oxide released from the injured endothelium. PMID- 11009466 TI - Methods for assessing hepatic distending pressure and changes in hepatic capacitance in pigs. AB - The equilibrium pressure obtained during simultaneous occlusion of hepatic vascular inflow and outflow was taken as the reference estimate of hepatic vascular distending pressure (P(hd)). P(hd) at baseline was 1.1 +/- 0.2 (mean +/- SE) mmHg higher than hepatic vein pressure (P(hv)) and 0.7 +/- 0.3 mmHg lower than portal vein pressure (P(pv)). Norepinephrine (NE) infusion increased P(hd) by 1. 5 +/- 0.5 mmHg and P(pv) by 3.7 +/- 0.6 mmHg but did not significantly increase P(hv). Hepatic lobar vein pressure (P(hlv)) measured by a micromanometer tipped 2-Fr catheter closely resembled P(hd) both at baseline and during NE infusion. Dynamic pressure-volume (PV) curves were constructed from continuous measurements of P(hv) and hepatic blood volume increases (estimated by sonomicrometry) during brief occlusions of hepatic vascular outflow and compared with static PV curves constructed from P(hd) determinations at five different hepatic volumes. Estimates of hepatic vascular compliance and changes in unstressed blood volume from the two methods were in close agreement with hepatic compliance averaging 32 +/- 2 ml. mmHg(-1). kg liver(-1). NE infusion reduced unstressed blood volume by 110 +/- 38 ml/kg liver but did not alter compliance. In conclusion, P(hlv) reflects hepatic distending pressure, and the construction of dynamic PV curves is a fast and valid method for assessing hepatic compliance and changes in unstressed blood volume. PMID- 11009467 TI - Angiotensin II acutely attenuates range of arterial baroreflex control of renal sympathetic nerve activity. AB - Acutely increasing peripheral angiotensin II (ANG II) reduces the maximum renal sympathetic nerve activity (RSNA) observed at low mean arterial blood pressures (MAPs). We postulated that this observation could be explained by the action of ANG II to acutely increase arterial blood pressure or increase circulating arginine vasopressin (AVP). Sustained increases in MAP and increases in circulating AVP have previously been shown to attenuate maximum RSNA at low MAP. In conscious rabbits pretreated with an AVP V1 receptor antagonist, we compared the effect of a 5-min intravenous infusion of ANG II (10 and 20 ng x kg(-1) x min(-1)) on the relationship between MAP and RSNA when the acute pressor action of ANG II was left unopposed with that when the acute pressor action of ANG II was opposed by a simultaneous infusion of sodium nitroprusside (SNP). Intravenous infusion of ANG II resulted in a dose-related attenuation of the maximum RSNA observed at low MAP. When the acute pressor action of ANG II was prevented by SNP, maximum RSNA at low MAP was attenuated, similar to that observed when ANG II acutely increased MAP. In contrast, intravertebral infusion of ANG II attenuated maximum RSNA at low MAP significantly more than when administered intravenously. The results of this study suggest that ANG II may act within the central nervous system to acutely attenuate the maximum RSNA observed at low MAP. PMID- 11009468 TI - Effects of mammary engorgement and feed withdrawal on microvascular function in lactating goat mammary glands. AB - The responses of the mammary microvasculature in lactating goats (n = 8) during feed withdrawal (18-20 h) and mammary engorgement (26-28 h of milk accumulation) were compared using an indicator-dilution technique with FITC-albumin and [(14)C]sucrose as the intravascular and diffusible indicators, respectively. Feed withdrawal and mammary engorgement caused a 50-60% decrease in mammary arterial flow and in the permeability-surface area product (PS) values for sucrose. Only feed withdrawal increased the mean transit time [from 17.3 to 30.0 s, SE of the difference (SED) = 2.16, P < 0.01] of FITC-albumin, whereas only mammary engorgement reduced sucrose extraction (0.63 to 0.51, SED = 0.04, P < 0.05). Mammary engorgement also caused a substantial reduction in the sucrose-accessible extravascular space from 92 to 44 ml (SED = 15.2, P < 0.01). In a separate experiment using five goats, milking after mammary engorgement did not immediately restore arterial flow or sucrose extraction, indicating that the effect of milk accumulation was not mediated simply via increased intramammary pressure. In conclusion, feed withdrawal resulted in slower flow in the capillary bed but apparently no change in capillary recruitment, whereas mammary engorgement caused capillary derecruitment. PMID- 11009469 TI - Role of pertussis toxin-sensitive G protein in metabolic vasodilation of coronary microcirculation. AB - We have previously demonstrated that pertussis toxin (PTX)-sensitive G protein (G(PTX)) plays a major role in coronary microvascular vasomotion during hypoperfusion. We aimed to elucidate the role of G(PTX) during increasing metabolic demand. In 18 mongrel dogs, coronary arteriolar diameters were measured by fluorescence microangiography using a floating objective. Myocardial oxygen consumption (MVO(2)) was increased by rapid left atrial pacing. In six dogs, PTX (300 ng/ml) was superfused onto the heart surface for 2 h to locally block G(PTX). In eight dogs, the vehicle (Krebs solution) was superfused in the same way. Before and after each treatment, the diameters were measured during control (130 beats/min) and rapid pacing (260 beats/min) in each group. Metabolic stimulation before and after the vehicle treatment caused 8.6 +/- 1. 8 and 16.1 +/- 3.6% dilation of coronary arterioles <100 microm in diameter (57 +/- 8 microm at control, n = 10), respectively. PTX treatment clearly abolished the dilation of arterioles (12.8 +/- 2. 5% before and 0.9 +/- 1.6% after the treatment, P < 0.001 vs. vehicle; 66 +/- 8 microm at control, n = 11) in response to metabolic stimulation. The increases in MVO(2) and coronary flow velocity were comparable between the vehicle and PTX groups. In four dogs, 8-phenyltheophylline (10 microM, superfusion for 30 min) did not affect the metabolic dilation of arterioles (15.3 +/- 2.0% before and 16.4 +/- 3.8% after treatment; 84.3 +/- 11.0 microm at control, n = 8). Thus we conclude that G(PTX) plays a major role in regulating the coronary microvascular tone during active hyperemia, and adenosine does not contribute to metabolic vasodilation via G(PTX) activation. PMID- 11009471 TI - 2,3-butanedione monoxime unmasks Ca(2+)-induced NADH formation and inhibits electron transport in rat hearts. AB - We used 2,3-butanedione monoxime (BDM) to suppress work by the perfused rat heart and to investigate the effects of calcium on NADH production and tissue energetics. Hearts were perfused with buffer containing BDM and elevated perfusate calcium to maintain the rates of cardiac work and oxygen consumption at levels similar to those of control perfused hearts. BDM plus calcium hearts displayed higher levels of NADH surface fluorescence, indicating calcium activation of mitochondrial dehydrogenases. These hearts, however, displayed 20% lower phosphocreatine levels. BDM suppressed the rates of state 3 respiration of isolated mitochondria. Uncoupled respiration was suppressed to a lesser degree, and the state 4 respiration rates were not affected. Double-inhibitor experiments with liver mitochondria using BDM and carboxyatractyloside (CAT) were used to identify the site of inhibition. BDM at low levels (0-5 mM) suppressed respiration. In the presence of CAT at levels that inhibit respiration by 60%, low levels of BDM were without effect. Because these effects were not additive, BDM does not inhibit adenine nucleotide transport. This was supported by an assay of adenine nucleotide transport in liver mitochondria. BDM did not inhibit ATP hydrolysis by submitochondrial particles but strongly suppressed reversed electron transport from succinate to NAD(+). Oxidation of NADH by submitochondrial particles was inhibited by BDM but oxidation of succinate was not. We conclude that BDM inhibits electron transport at site 1. PMID- 11009470 TI - Relationship of molecular structure to the mechanism of lysophospholipid-induced smooth muscle cell proliferation. AB - We previously reported that oxidized low-density lipoprotein and one of its constituents, lysophosphatidylcholine (lysoPC), caused smooth muscle cell proliferation that was inhibitable by vitamin E and by a neutralizing antibody against basic fibroblast growth factor-2 (FGF-2). We now show that the mitogenic activity of lysolipids is highly dependent on structure. Phospholipids with palmitoyl fatty acid and phosphocholine induced DNA synthesis optimally. Shorter and longer fatty acids were significantly less potent, as were phosphoserine and phosphoethanolamine head groups. Structurally related phospholipids [platelet activating factor (PAF) and lysoPAF] were also mitogens and acted via an analogous FGF-2-dependent, vitamin E-inhibitable mechanism. The mechanism of lysoPC stimulation was distinct from that of another phospholipid mitogen, lysophosphatidic acid (lysoPA), in that lysoPC stimulation was not pertussis toxin inhibitable. Furthermore, lysoPA stimulation was not inhibitable by vitamin E. Despite its distinct cellular pathway for stimulation, lysoPA also ultimately led to FGF-2 release. Our data show that specific structural attributes of lysoPC, PAF, and lysoPAF enable these agents to mediate smooth muscle cell release of FGF-2, which in turn stimulates proliferation. PMID- 11009472 TI - Mitochondrial NAD(P)H, ADP, oxidative phosphorylation, and contraction in isolated heart cells. AB - To examine the relationship between mitochondrial NADH (NADH(m)) and cardiac work output, NADH(m) and the amplitude and frequency of the contractile response of electrically paced rat heart cells were measured at 25 degrees C. With 5.4 mM glucose plus 2 mM beta-hydroxybutyrate, NADH(m) was reversibly decreased by 23%, and the amplitude of contraction was reversibly decreased by 27% during 4-Hz pacing. With glucose plus 2 mM pyruvate or with 10 mM 2-deoxy-D-glucose, NADH(m) was maintained during rapid pacing, and the contractile amplitude remained high. Phosphocreatine levels decreased with 2-deoxy-D-glucose administration but not with rapid pacing. Respiration increased to meet the increased ATP demand at 30 degrees C. The data suggest that 1) when NADH(m) is decreased during rapid pacing with defined substrates, the amplitude of contraction is decreased; 2) the amplitude of contraction during electrical pacing does not change with rate of pacing when both the ATP and NADH(m) levels are continuously replenished; and 3) the replenishment of NADH(m) during pacing with physiological substrates may be rate-limited by substrate supply to mitochondrial dehydrogenases. During activation of mitochondrial dehydrogenases, or a significant increase in free ADP induced by 2-deoxy-D-glucose, this rate limitation is bypassed or overcome. PMID- 11009477 TI - Prior exercise training increases collateral-dependent blood flow in rats after acute femoral artery occlusion. AB - We evaluated whether prior training would improve collateral blood flow (BF) to the calf muscles after acute-onset occlusion of the femoral artery. Exercise training was performed in the absence of any vascular occlusion. Adult male Sprague-Dawley rats ( approximately 325 g) were kept sedentary (n = 14), limited to cage activity, or exercise trained (n = 14) for 6 wk by treadmill running. Early in the day of measurement, animals were surgically prepared for BF determination, and the femoral arteries were occluded bilaterally. Four to five hours later, collateral BF was determined twice during treadmill running with the use of (141)Ce and (85)Sr microspheres: first, at a demanding speed and, second, after a brief rest and at a higher speed. The absence of any further increase in BF at the higher speed indicated that maximal collateral BF was measured. Prior training increased calf muscle BF by approximately 70% compared with sedentary animals; however, absolute BF remained below values previously observed in animals with a well-developed collateral vascular tree. Thus prior training appeared to optimize the use of the existing collateral circuit. This implies that altered vasoresponsiveness induced in normal nonoccluded vessels with exercise training serves to improve collateral BF to the periphery. PMID- 11009475 TI - Dynamics of action potential head-tail interaction during reentry in cardiac tissue: ionic mechanisms. AB - In a sufficiently short reentry pathway, the excitation wave front (head) propagates into tissue that is partially refractory (tail) from the previous action potential (AP). We incorporate a detailed mathematical model of the ventricular myocyte into a one-dimensional closed pathway to investigate the effects of head-tail interaction and ion accumulation on the dynamics of reentry. The results were the following: 1) a high degree of head-tail interaction produces oscillations in several AP properties; 2) Ca(2+)-transient oscillations are in phase with AP duration oscillations and are often of greater magnitude; 3) as the wave front propagates around the pathway, AP properties undergo periodic spatial oscillations that produce complicated temporal oscillations at a single site; 4) depending on the degree of head-tail interaction, intracellular [Na(+)] accumulation during reentry either stabilizes or destabilizes reentry; and 5) elevated extracellular [K(+)] destabilizes reentry by prolonging the tail of postrepolarization refractoriness. PMID- 11009473 TI - Flumazenil preconditions cardiomyocytes via oxygen radicals and K(ATP) channels. AB - We determined whether flumazenil mimics ischemic preconditioning in chick cardiomyocytes and examined the role of intracellular reactive oxygen species (ROS) and ATP-dependent potassium (K(ATP)) channels in mediating the effect. Chick ventricular myocytes were perfused with a balanced salt solution in a flow through chamber. Cell viability was quantified using propidium iodide, and ROS generation was assessed using the reduced form of 2',7'-dichlorofluorescin (DCFH). Cells were exposed to 1 h of simulated ischemia and 3 h of reoxygenation. Preconditioning was initiated with 10 min of ischemia followed by 10 min of reoxygenation. Alternatively, flumazenil was added to the perfusate for 10 min and removed 10 min before the start of ischemia. Flumazenil (1 and 10 microM) and preconditioning reduced cell death [54 +/- 5%, n = 3; 26 +/- 4%, n = 6 (P < 0.05); and 20 +/- 2%, n = 6 (P < 0.05), respectively, vs. 57 +/- 7%, n = 10, in controls] and increased DCFH oxidation (an index of ROS production) [0.35 +/- 0.11, n = 3; 2.64 +/- 0.69, n = 8 (P < 0.05); and 2.46 +/- 0.52, n = 6 (P < 0.05), respectively, vs. 0.26 +/- 0.05, n = 9, in controls]. Protection and increased ROS signals with flumazenil (10 microM) were abolished with the thiol reductant N-(2-mercaptopropionyl)-glycine (2-MPG, 800 microM), an antioxidant (cell death: 2-MPG + flumazenil, 55 +/- 12%, n = 6; ROS signals: 2-MPG + flumazenil, 0.11 +/- 0.19, n = 6). Treatment with 5-hydroxydecanoate (1 mM), a selective mitochondrial K(ATP) channel antagonist, abolished its protection. These results demonstrate that flumazenil mimics preconditioning to reduce cell death in myocytes. ROS signals with the resultant mitochondrial K(ATP) channel activation are important components of the intracellular signaling pathway of flumazenil. PMID- 11009474 TI - Protein washdown as a defense mechanism against myocardial edema. AB - Myocardial edema occurs in many pathological conditions. We hypothesized that protein washdown at the myocardial microvascular exchange barrier would change the distribution of interstitial proteins from large to small molecules and diminish the effect of washdown on the colloid osmotic pressure (COP) of interstitial fluid and lymph. Dogs were instrumented with coronary sinus balloon tipped catheters and myocardial lymphatic cannulas to manipulate myocardial lymph flow and to collect lymph. Myocardial venous pressure was elevated by balloon inflation to increase transmicrovascular fluid flux and myocardial lymph flow. COP of lymph was measured directly and was also calculated from protein concentration. Decreases occurred in both protein concentration and COP of lymph. The proportion of lymph protein accounted for by albumin increased significantly, whereas that accounted for by beta-lipoprotein decreased significantly. The change in the calculated plasma-to-lymph COP gradient was significantly greater than the change in the measured COP gradient. We conclude that the change in the distribution of interstitial fluid protein species decreases the effect of protein washdown on interstitial fluid COP and limits its effectiveness as a defense mechanism against myocardial edema formation. PMID- 11009478 TI - Effect of milrinone on left ventricular relaxation and Ca(2+) uptake function of cardiac sarcoplasmic reticulum. AB - Milrinone, a phosphodiesterase 3 (PDE3) inhibitor, is known to enhance left ventricular (LV) contractility by an inhibition of the breakdown of cAMP through the mechanism inhibiting PDE3. However, it is unclear whether milrinone also exerts positive lusitropy, like dobutamine. Here, we assessed the effects of milrinone on in vivo LV relaxation, as well as the Ca(2+)-ATPase activity and the Ca(2+) uptake function of the cardiac sarcoplasmic reticulum (SR), compared with the effect of dobutamine on those functions. After dobutamine (3 microg x kg(-1) x min(-1)) was administered, the peak value of the first derivative of LV pressure (+dP/dt) increased by 46%, whereas the time constant (tau) of LV pressure decay decreased by 6.9%, respectively. After milrinone (10 microg/kg) was administered, the peak +dP/dt increased to a similar extent as dobutamine (46%), whereas tau decreased much more than dobutamine (19.9%; P < 0.05). In LV crude homogenate, the thapsigargin-sensitive, Ca(2+)-ATPase activity-cAMP relationships was significantly less increased by milrinone compared with dobutamine (P < 0.05), indicating the higher sensitivity of the SR Ca(2+)-ATPase activity on cAMP by milrinone than by dobutamine. In the SR vesicles purified from LV muscles, the addition of cAMP increased the SR Ca(2+) uptake in a dose dependent fashion, and the PDE3 inhibitors (milrinone and cGMP) significantly augmented this response (P < 0.05). Hence, milrinone substantially improved LV relaxation in association with an acceleration of the SR Ca(2+)-ATPase activity and the SR Ca(2+) uptake. This acceleration might be due to an inhibition of the membrane-bound PDE3 in the SR, leading to a local elevation of cAMP. PMID- 11009476 TI - Interactions of hemoglobin with hydrogen peroxide alters thiol levels and course of endothelial cell death. AB - We investigated cellular injury and death induced by ultrapure human Hb (HbA(0)) and its diaspirin cross-linked derivative DBBF-Hb in normal and glutathione (GSH) depleted bovine aortic endothelial cells subjected to hydrogen peroxide (H(2)O(2)). HbA(0) underwent extensive degradation and heme loss, whereas DBBF-Hb persisted longer in its ferryl (Fe(4+)) form. The formation of ferryl HbA(0) or ferryl DBBF-Hb was associated with a significant decrease in endothelial cell GSH compared with the addition of H(2)O(2) or Hbs alone. This effect was inhibited by catalase, but not by superoxide dismutase or deferoxamine mesylate. The presence of HbA(0) and DBBF-Hb reduced H(2)O(2)-induced apoptosis, as measured by cell morphology, annexin V binding assay, and caspase inhibition, consistent with the ability to consume H(2)O(2) in an enzyme-like fashion. However, the pattern of cell death and injury produced by HbA(0) and DBBF-Hb appeared to be distinctly different among proteins as well as among cells with and without GSH. These findings may have important implications for the use of cell-free Hb as oxygen therapeutics in patients with coexisting pathologies who may lack antioxidant protective mechanisms. PMID- 11009479 TI - Vasodilator mechanisms in the coronary circulation of endothelial nitric oxide synthase-deficient mice. AB - Previous studies have demonstrated that responses to endothelium-dependent vasodilators are absent in the aortas from mice deficient in expression of endothelial nitric oxide synthase (eNOS -/- mice), whereas responses in the cerebral microcirculation are preserved. We tested the hypothesis that in the absence of eNOS, other vasodilator pathways compensate to preserve endothelium dependent relaxation in the coronary circulation. Diameters of isolated, pressurized coronary arteries from eNOS -/-, eNOS heterozygous (+/-), and wild type mice (eNOS +/+ and C57BL/6J) were measured by video microscopy. ACh (an endothelium-dependent agonist) produced vasodilation in wild-type mice. This response was normal in eNOS +/- mice and was largely preserved in eNOS -/- mice. Responses to nitroprusside were also similar in arteries from eNOS +/+, eNOS +/-, and eNOS -/- mice. Dilation to ACh was inhibited by N(G)-nitro-L-arginine, an inhibitor of NOS in control and eNOS -/- mice. In contrast, trifluoromethylphenylimidazole, an inhibitor of neuronal NOS (nNOS), decreased ACh-induced dilation in arteries from eNOS-deficient mice but had no effect on responses in wild-type mice. Indomethacin, an inhibitor of cyclooxygenase, decreased vasodilation to ACh in eNOS-deficient, but not wild-type, mice. Thus, in the absence of eNOS, dilation of coronary arteries to ACh is preserved by other vasodilator mechanisms. PMID- 11009480 TI - Time and dose effect of transdermal nicotine on endothelial function. AB - Nicotine patches are available as an over-the-counter medication for aid in smoking cessation. This study was designed to determine how nicotine patch therapy over time and dose ranges used in smoking cessation programs in humans affects endothelium-dependent relaxations. Dogs were treated with nicotine patches (11, 22, or 44 mg/day) for 2 and 5 wk. Circulating nicotine and oxidized products of nitric oxide (NOx) were measured. Coronary arteries were prepared for measurement of isometric force and aortic endothelial cells were prepared for measurement of mRNA or NO synthase (NOS) activity. Circulating nicotine increased with increasing concentrations of nicotine patches. After 5 wk of treatment with 22 mg/day patches, circulating NOx was reduced but NOS activity was increased. NOS mRNA was similar among groups. Only after 5 wk of treatment with 22 mg/day patches were endothelium-dependent relaxations reduced to alpha(2)-adrenergic agonists, ADP, and the calcium ionophore A-23187. These results suggest a time and biphasic dose effect of nicotine treatment on endothelium-dependent responses that may be related to bioavailability of NO. This complex relationship of duration and dose of nicotine treatment may explain, in part, discrepancies in effects of nicotine on endothelium-dependent responses. PMID- 11009481 TI - Diaspirin cross-linked Hb and norepinephrine prevent the sepsis-induced increase in critical O(2) delivery. AB - We hypothesized that support of arterial perfusion pressure with diaspirin cross linked Hb (DCLHb) would prevent the sepsis-induced attenuation in the systemic O(2) delivery-O(2) uptake relationship. Awake septic rats were treated with a chronic infusion of DCLHb or a reference treatment [norepinephrine (NE)] to increase mean arterial pressure by 10-20% over 18 h. Septic and sham control groups received normal saline. Isovolemic hemodilution to create anemic hypoxia was then performed in a metabolic box during continuous measurement of systemic O(2) uptake. O(2) delivery was calculated from hemodynamic variables, and the critical point of O(2) delivery (DO(2 crit)) was determined using piecewise regression analysis of the O(2) delivery-O(2) uptake relationship. Sepsis increased DO(2 crit) from 4.99 +/- 0.17 to 6.69 +/- 0.42 ml x min(-1) x 100 g(-1) (P < 0.01), while O(2) extraction capacity was decreased (P < 0.05). DCLHb and NE infusion prevented the sepsis-induced increase in DO(2 crit) [4.56 +/- 0.42 ml x min(-1) x 100 g(-1) (P < 0.01) and 5.04 +/- 0.56 ml x min(-1) x 100 g(-1) (P < 0.05), respectively]. This was explained by a 59% increase in O(2) extraction capacity in the DCLHb group compared with septic controls (P < 0.05), whereas NE treatment decreased systemic O(2) uptake in anemic hypoxia (1.51 +/- 0.08 vs. 1.87 +/- 0.1 ml x min(-1) x 100 g(-1) in septic controls, P < 0.05). We conclude that DCLHb ameliorated O(2) extraction capacity in the septic microcirculation, whereas NE decreased the metabolic demands of the tissues. PMID- 11009482 TI - Cardiovascular and neuroendocrine responses to water immersion in compensated heart failure. AB - The hypothesis was tested that cardiovascular and neuroendocrine (norepinephrine, renin, and vasopressin) responses to central blood volume expansion are blunted in compensated heart failure (HF). Nine HF patients [New York Heart Association class II-III, ejection fraction = 0.28 +/- 0.02 (SE)] and 10 age-matched controls (ejection fraction = 0.68 +/- 0.03) underwent 30 min of thermoneutral (34.7 +/- 0.02 degrees C) water immersion (WI) to the xiphoid process. WI increased (P < 0.05) central venous pressure by 3.7 +/- 0.6 and 3.2 +/- 0.4 mmHg and stroke volume index by 12.2 +/- 2.1 and 7.2 +/- 2.1 ml. beat(-1). m(-2) in controls and HF patients, respectively. During WI, systemic vascular resistance decreased (P < 0.05) similarly by 365 +/- 66 and 582 +/- 227 dyn. s. cm(-5) in controls and HF patients, respectively. Forearm subcutaneous vascular resistance decreased by 19 +/- 7% (P < 0.05) in controls but did not change in HF patients. Heart rate decreased less during WI in HF patients, whereas release of norepinephrine, renin, and vasopressin was suppressed similarly in the two groups. We suggest that reflex control of forearm vascular beds and heart rate is blunted in compensated HF but that baroreflex-mediated systemic vasodilatation and neuroendocrine responses to central blood volume expansion are preserved. PMID- 11009483 TI - Cardioprotection with kappa-opioid receptor stimulation is associated with a slowing of cross-bridge cycling. AB - Opioid and alpha-adrenergic receptor activation protect the heart from ischemic damage. One possible intracellular mechanism to explain this is that an improvement in ATP availability contributes to cardioprotection. We tested this hypothesis by correlating postischemic left ventricular developed pressure (LVDP) and myofibrillar Ca(2+)-dependent actomyosin Mg(2+)-ATPase from isolated rat hearts treated with the kappa-opioid receptor agonist U-50488H (1 microM) or the alpha-adrenergic receptor agonist phenylephrine (10 microM) + propranolol (3 microM). Preischemic treatment with U-50488H or phenylephrine + propranolol improved postischemic LVDP recovery by 25-30% over control hearts. Ca(2+) dependent actomyosin Mg(2+)-ATPase was found to be 20% lower in both U-50488H- and phenylephrine + propranolol-treated hearts compared with control hearts. The kappa-opioid receptor antagonist nor-binaltorphimine (1 microM) abolished the effects of U-50488H on postischemic LVDP and actomyosin Mg(2+)-ATPase activity. Reduced actomyosin ATP utilization was also suggested in single ventricular myocytes treated with either U-50488H or the protein kinase C activator, phorbol 12-myristate 13-acetate (PMA), because U-50488H and PMA lowered maximum velocity of unloaded shortening by 15-25% in myocytes. U-50488H and phenylephrine + propranolol treatment both resulted in increased phosphorylation of troponin I and C protein. These findings are consistent with the hypothesis that kappa opioid and alpha-adrenergic receptors decrease actin-myosin cycling rate, leading to a conservation of ATP and cardioprotection during ischemia. PMID- 11009484 TI - Plasma viscosity and cerebral blood flow. AB - We hypothesized that the response of cerebral blood flow (CBF) to changing viscosity would be dependent on "baseline" CBF, with a greater influence of viscosity during high-flow conditions. Plasma viscosity was adjusted to 1.0 or 3.0 cP in rats by exchange transfusion with red blood cells diluted in lactated Ringer solution or with dextran. Cortical CBF was measured by H(2) clearance. Two groups of animals remained normoxic and normocarbic and served as controls. Other groups were made anemic, hypercapnic, or hypoxic to increase CBF. Under baseline conditions before intervention, CBF did not differ between groups and averaged 49.4 +/- 10.2 ml. 100 g(-1). min(-1) (+/-SD). In control animals, changing plasma viscosity to 1. 0 or 3.0 cP resulted in CBF of 55.9 +/- 8.6 and 42.5 +/- 12.7 ml. 100 g(-1). min(-1), respectively (not significant). During hemodilution, hypercapnia, and hypoxia with a plasma viscosity of 1. 0 cP, CBF varied from 98 to 115 ml. 100 g(-1). min(-1). When plasma viscosity was 3.0 cP during hemodilution, hypercapnia, and hypoxia, CBF ranged from 56 to 58 ml. 100 g(-1). min(-1) and was significantly reduced in each case (P < 0.05). These results support the hypothesis that viscosity has a greater role in regulation of CBF when CBF is increased. In addition, because CBF more closely followed changes in plasma viscosity (rather than whole blood viscosity), we believe that plasma viscosity may be the more important factor in controlling CBF. PMID- 11009485 TI - Carotid baroreflex responsiveness in heat-stressed humans. AB - The effects of whole body heating on human baroreflex function are relatively unknown. The purpose of this project was to identify whether whole body heating reduces the maximal slope of the carotid baroreflex. In 12 subjects, carotid vasomotor and carotid-cardiac baroreflex responsiveness were assessed in normothermia and during whole body heating. Whole body heating increased sublingual temperature (from 36.4 +/- 0.1 to 37.4 +/- 0.1 degrees C, P < 0.01) and increased heart rate (from 59 +/- 3 to 83 +/- 3 beats/min, P < 0. 01), whereas mean arterial blood pressure (MAP) was slightly decreased (from 88 +/- 2 to 83 +/- 2 mmHg, P < 0.01). Carotid-vasomotor and carotid-cardiac responsiveness were assessed by identifying the maximal gain of MAP and heart rate to R wave triggered changes in carotid sinus transmural pressure. Whole body heating significantly decreased the responsiveness of the carotid-vasomotor baroreflex (from -0.20 +/- 0.02 to -0.13 +/- 0.02 mmHg/mmHg, P < 0.01) without altering the responsiveness of the carotid-cardiac baroreflex (from -0.40 +/- 0.05 to -0.36 +/ 0.02 beats x min(-1) x mmHg(-1), P = 0.21). Carotid-vasomotor and carotid cardiac baroreflex curves were shifted downward and upward, respectively, to accommodate the decrease in blood pressure and increase in heart rate that accompanied the heat stress. Moreover, the operating point of the carotid-cardiac baroreflex was shifted closer to threshold (P = 0.02) by the heat stress. Reduced carotid-vasomotor baroreflex responsiveness, coupled with a reduction in the functional reserve for the carotid baroreflex to increase heart rate during a hypotensive challenge, may contribute to increased susceptibility to orthostatic intolerance during a heat stress. PMID- 11009486 TI - Biphasic effects of hyposmotic challenge on excitation-contraction coupling in rat ventricular myocytes. AB - The effects of short (1 min) and long (7-10 min) exposure to hyposmotic solution on excitation-contraction coupling in rat ventricular myocytes were studied. After short exposure, the action potential duration at 90% repolarization (APD(90)), the intracellular Ca(2+) concentration ([Ca(2+)](i)) transient amplitude, and contraction increased, whereas the L-type Ca(2+) current (I(Ca, L)) amplitude decreased. Fractional sarcoplasmic reticulum (SR) Ca(2+) release increased but SR Ca(2+) load did not. After a long exposure, I(Ca,L), APD(90), [Ca(2+)](i) transient amplitude, and contraction decreased. The abbreviation of APD(90) was partially reversed by 50 microM DIDS, which is consistent with the participation of Cl(-) current activated by swelling. After 10-min exposure to hyposmotic solution in cells labeled with di-8-aminonaphthylethenylpyridinium, t tubule patterning remained intact, suggesting the loss of de-t-tubulation was not responsible for the fall in I(Ca,L). After long exposure, Ca(2+) load of the SR was not increased, and swelling had no effect on the site-specific phosphorylation of phospholamban, but fractional SR Ca(2+) release was depressed. The initial positive inotropic response to hyposmotic challenge may be accounted for by enhanced coupling between Ca(2+) entry and release. The negative inotropic effect of prolonged exposure can be accounted for by shortening of the action potential duration and a fall in the I(Ca,L) amplitude. PMID- 11009487 TI - Na(+)-K(+)-ATPase alpha(2)-isoform expression in guinea pig hearts during transition from compensation to decompensation. AB - Disturbance in ionic gradient across sarcolemma may lead to arrhythmias. Because Na(+)-K(+)-ATPase regulates intracellular Na(+) and K(+) concentrations, and therefore intracellular Ca(2+) concentration homeostasis, our aim was to determine whether changes in the Na(+)-K(+)-ATPase alpha-isoforms in guinea pigs during transition from compensated (CLVH) to decompensated left ventricular hypertrophy (DLVH) were concomitant with arrhythmias. After 12- and 20-mo aortic stenosis, CLVH and DLVH were characterized by increased mean arterial pressure (30% and 52.7%, respectively). DLVH differed from CLVH by significantly increased end-diastolic pressure (34%), decreased sarco(endo)plasmic reticulum Ca(2+) ATPase (-75%), and increased Na(+)/Ca(2+) exchanger (25%) mRNA levels and by the occurrence of ventricular arrhythmias. The alpha-isoform (mRNA and protein levels) was significantly lower in DLVH (2.2 +/- 0.2- and 1. 4 +/- 0.15-fold, respectively, vs. control) than in CLVH (3.5 +/- 0. 4- and 2.2 +/- 0.13-fold, respectively) and was present in sarcolemma and T tubules. Changes in the levels of alpha(1)- and alpha(3)-isoform in CLVH and DLVH appear physiologically irrelevant. We suggest that the increased level of alpha(2)-isoform in CLVH may participate in compensation, whereas its relative decrease in DLVH may enhance decompensation and arrhythmias. PMID- 11009488 TI - cGMP-independent inotropic effects of nitric oxide and peroxynitrite donors: potential role for nitrosylation. AB - Nitric oxide (NO) has concentration-dependent biphasic myocardial contractile effects. We tested the hypothesis, in isolated rat hearts, that NO cardiostimulation is primarily non-cGMP dependent. Infusion of 3 morpholinosydnonimine (SIN-1, 10(-5) M), which may participate in S-nitrosylation (S-NO) via peroxynitrite formation, increased the rate of left ventricular pressure rise (+dP/dt; 19 +/- 4%, P < 0.001, n = 11) without increasing effluent cGMP or cAMP. Superoxide dismutase (SOD; 150 U/ml) blocked SIN-1 cardiostimulation and led to cGMP elaboration. Sodium nitroprusside (10(-10)-10( 7) M), an iron nitrosyl compound, did not augment +dP/dt but increased cGMP approximately eightfold (P < 0.001), whereas diethylamine/NO (DEA/NO; 10(-7) M), a spontaneous NO. donor, increased +dP/dt (5 +/- 2%, P < 0.05, n = 6) without augmenting cGMP. SIN-1 and DEA/NO +dP/dt increase persisted despite guanylyl cyclase inhibition with 1H-(1,2,4)oxadiazolo-(4,3,-a)quinoxalin-1-one (10(-5) M, P < 0.05 for both donors), suggesting a cGMP-independent mechanism. Glutathione (5 x 10(-4) M, n = 15) prevented SIN-1 cardiostimulation, suggesting S-NO formation. SIN-1 also produced SOD-inhibitable cardiostimulation in vivo in mice. Thus peroxynitrite and NO donors can stimulate myocardial contractility independently of guanylyl cyclase activation, suggesting a role for S-NO reactions in NO/peroxynitrite-positive inotropic effects in intact hearts. PMID- 11009489 TI - Cardiac synthesis, processing, and coronary release of enkephalin-related peptides. AB - Although preproenkephalin mRNA is abundant in the heart, the myocardial synthesis and processing of proenkephalin is largely undefined. Isolated working rat hearts were perfused to determine the rate of myocardial proenkephalin synthesis, its processing into enkephalin-containing peptides, their subsequent release into the coronary arteries, and the influence of prior sympathectomy. Enkephalin containing peptides were separated by gel filtration and quantified with antisera for specific COOH-terminal sequences. Proenkephalin, peptide B, and [Met(5)]enkephalin-Arg(6)-Phe(7) (MEAP) comprised 95% of the extracted myocardial enkephalins (35 pmol/g). Newly synthesized enkephalins, estimated during a 1-h perfusion with [(14)C]phenylalanine (4 pmol x h(-1) x g wet wt(-1)), were rapidly cleared from the heart during a second isotope-free hour. Despite a steady release of enkephalins into the coronary effluent (4 pmol x h(-1) x g wet wt( 1)), enkephalin replacement apparently exceeded its release, and tissue enkephalins actually accumulated during hour 2. In contrast to the tissue, methionine-enkephalin accounted for more than half of the released enkephalin. Chemical sympathectomy produced an increase in total enkephalin content similar to that observed after 2-h control perfusion. This observation suggested that the normal turnover of myocardial enkephalin may depend in part on continued sympathetic influences. PMID- 11009490 TI - Effect of aerobic and resistance exercise training on vascular function in heart failure. AB - Exercise training of a muscle group improves local vascular function in subjects with chronic heart failure (CHF). We studied forearm resistance vessel function in 12 patients with CHF in response to an 8-wk exercise program, which specifically excluded forearm exercise, using a crossover design. Forearm blood flow (FBF) was measured using strain-gauge plethysmography. Responses to three dose levels of intra-arterial acetylcholine were significantly augmented after exercise training when analyzed in terms of absolute flows (7.0 +/- 1.8 to 10.9 +/- 2.1 ml x 100 ml(-1) x min(-1) for the highest dose, P < 0.05 by ANOVA), forearm vascular resistance (21.5 +/- 5.0 to 15.3 +/- 3.9 ml x 100 ml forearm(-1) x min(-1), P < 0.01), or FBF ratios (P < 0.01, ANOVA). FBF ratio responses to sodium nitroprusside were also significantly increased after training (P < 0.05, ANOVA). Reactive hyperemic flow significantly increased in both upper limbs after training (27.9 +/- 2.7 to 33.5 +/- 3.1 ml x 100 ml(-1) x min(-1), infused limb; P < 0.05 by paired t-test). Exercise training improves endothelium-dependent and independent vascular function and peak vasodilator capacity in patients with CHF. These effects on the vasculature are generalized, as they were evident in a vascular bed not directly involved in the exercise stimulus. PMID- 11009492 TI - Echocardiographic and invasive measurements of pulmonary artery pressure correlate closely at high altitude. AB - Exaggerated hypoxia-induced pulmonary hypertension is a hallmark of high-altitude pulmonary edema (HAPE) and plays a major role in its pathogenesis. Many studies of HAPE have estimated systolic pulmonary arterial pressure (SPAP) with Doppler echocardiography. Whereas at low altitude, Doppler echocardiographic estimation of SPAP correlates closely with its invasive measurement, no such evidence exists for estimations obtained at high altitude, where alterations of blood viscosity may invalidate the simplified Bernoulli equation. We measured SPAP by Doppler echocardiography and invasively in 14 mountaineers prone to HAPE and in 14 mountaineers resistant to this condition at 4,559 m. Mountaineers prone to HAPE had more pronounced pulmonary hypertension (57 +/- 12 and 58 +/- 10 mmHg for noninvasive and invasive determination, respectively; means +/- SD) than subjects resistant to HAPE (37 +/- 8 and 37 +/- 6 mmHg, respectively), and the values measured in the two groups as a whole covered a wide range of pulmonary arterial pressures (30-83 mmHg). Spearman test showed a highly significant correlation (r = 0.89, P < 0.0001) between estimated and invasively measured SPAP values. The mean difference between invasively measured and Doppler-estimated SPAP was 0.5 +/ 8 mmHg. At high altitude, estimation of SPAP by Doppler echocardiography is an accurate and reproducible method that correlates closely with its invasive measurement. PMID- 11009491 TI - Evidence for nitroxidergic innervation in monkey ophthalmic arteries in vivo and in vitro. AB - In anesthetized monkeys, electrical stimulation (ES) of the pterygopalatine or geniculate ganglion dilated the ipsilateral ophthalmic artery (OA). The induced vasodilatation was unaffected by phentolamine but potentiated by atropine. Intravenous N(G)-nitro-L-arginine (L-NNA) abolished the response, which was restored by L-arginine. Hexamethonium-abolished vasodilator responses induced solely by geniculate ganglionic stimulation. The L-NNA constricted OA; L-arginine reversed the effect. Destruction of the pterygopalatine ganglion constricted the ipsilateral artery. Helical strips of OA isolated under deep anesthesia from monkeys, denuded of endothelium, responded to transmural ES with relaxations, which were abolished by tetrodotoxin and L-NNA but were potentiated by atropine. It is concluded that neurogenic vasodilatation of monkey OA is mediated by nerve derived nitric oxide (NO), and the nerve is originated from the ipsilateral pterygopalatine ganglion that is innervated by cholinergic neurons from the brain stem via the geniculate ganglion. The OA appears to be dilated by mediation of NO continuously liberated from nerves that receive tonic discharges from the vasomotor center. Acetylcholine liberated from postganglionic cholinergic nerves would impair the release of neurogenic NO. PMID- 11009493 TI - Differential effects of L-NAME on rat venular hydraulic conductivity. AB - The role of nitric oxide (NO) in microvascular permeability remains unclear because both increases and decreases in permeability by NO synthase (NOS) inhibitors have been reported. We sought to determine whether blood-borne constituents modify venular permeability responses to the NOS inhibitor N(G) nitro-L-arginine methyl ester (L-NAME). We assessed hydraulic conductivity (L(p)) of pipette-perfused rat mesenteric venules before and after exposure to 10(-4) M L-NAME. In the absence of blood-borne constituents, L-NAME reduced L(p) by nearly 50% (from a median of 2.4 x 10(-7) cm x s(-1) x cmH(2)O(-1), n = 17, P < 0.001). The reduction in L(p) by L-NAME was inhibited by a 10-fold molar excess of L arginine but not D-arginine (n = 6). In a separate group of venules, blood flow was allowed to resume during exposure to L-NAME. In vessels perfused by blood during L-NAME exposure, L(p) increased by 78% (from 1.4 x 10(-7) cm x s(-1) x cmH(2)O(-1), n = 10, P < 0.01). N(G)-nitro-D-arginine methyl ester did not affect L(p) in either of the two groups. These data imply that NO has direct vascular effects on permeability that are opposed by secondary changes in permeability mediated by blood-borne constituents. PMID- 11009494 TI - Voltage-independent changes in L-type Ca(2+) current uncoupled from SR Ca(2+) release in cardiac myocytes. AB - To determine the effect of voltage-independent alterations of L-type Ca(2+) current (I(Ca)) on the sarcoplasmic reticular (SR) Ca(2+) release in cardiac myocytes, we measured I(Ca) and cytosolic Ca(2+) transients (Ca(i)(2+); intracellular Ca(2+) concentration) in voltage-clamped rat ventricular myocytes during 1) an abrupt increase of extracellular [Ca(2+)] (Ca(o)(2+)) or 2) application of 1 microM FPL-64176, a Ca(2+) channel agonist, to selectively alter I(Ca) in the absence of changes in SR Ca(2+) loading. On the first depolarization in higher Ca(o)(2+), peak I(Ca) was increased by 46 +/- 6% (P < 0.001), but the increases in the maximal rate of rise of Ca(i)(2+) (dCa(i)(2+)/dt(max), where t is time; an index of SR Ca(2+) release flux) and the Ca(i)(2+) transient amplitude were not significant. Rapid exposure to FPL-64176 greatly slowed inactivation of I(Ca), increasing its time integral by 117 +/- 8% (P < 0.001) without significantly increasing peak I(Ca), dCa(i)(2+)/dt(max), or amplitude of the corresponding Ca(i)(2+) transient. Prolongation of exposure to higher Ca(o)(2+) or FPL-64176 did not further increase peak I(Ca) but greatly increased dCa(i)(2+)/dt(max), Ca(i)(2+) transient amplitude, and the gain of Ca(2+) release (dCa(i)(2+)/dt(max)/I(Ca)), evidently due to augmentation of the SR Ca(2+) loading. Also, the time to peak dCa(i)(2+)/dt(max) was significantly increased in the continuous presence of higher Ca(o)(2+) (by 37 +/- 5%, P < 0.001) or FPL 64176 (by 63 +/- 5%, P < 0.002). Our experiments provide the first evidence of a marked disparity between an increased peak I(Ca) and the corresponding SR Ca(2+) release. We attribute this to saturation of the SR Ca(2+) release flux as predicted by local control theory. Prolongation of the SR Ca(2+) release flux, caused by combined actions of a larger I(Ca) and maximally augmented SR Ca(2+) loading, might reflect additional Ca(2+) release from corbular SR. PMID- 11009495 TI - Strengthening governance for global health research. PMID- 11009496 TI - Global information flow. PMID- 11009497 TI - The ethics of international biomedical research. PMID- 11009499 TI - Why the human rights act matters to doctors. PMID- 11009498 TI - The management of post-herpetic neuralgia. PMID- 11009500 TI - Testicular cancer and infertility. PMID- 11009502 TI - US prescription drug sales boosted by advertising. PMID- 11009501 TI - The separating of conjoined twins. PMID- 11009503 TI - Conjoined twins should be separated. PMID- 11009504 TI - Laparoscopic surgery: two thirds of injuries initially missed. PMID- 11009505 TI - Shipman inquiry to be held in public. PMID- 11009506 TI - In brief PMID- 11009507 TI - Mammography is no better than physical examination, study shows PMID- 11009508 TI - Public criticism makes paediatric surgeon's job too difficult. PMID- 11009509 TI - New sweet potato could help combat blindness in Africa. PMID- 11009510 TI - Government policies set to narrow "health gap". PMID- 11009511 TI - Global health agencies are accused of incompetence. PMID- 11009512 TI - WHO links with consumers on reproductive health information. PMID- 11009513 TI - AIDS vaccine research focuses on subtypes in developed world. PMID- 11009514 TI - Toiling in tough times. PMID- 11009515 TI - Risk of testicular cancer in men with abnormal semen characteristics: cohort study. AB - OBJECTIVE: To explore the associations between semen characteristics and subsequent risk of testicular cancer. DESIGN: Cohort study. PARTICIPANTS: 32 442 men who had a semen analysis done at the Sperm Analysis Laboratory in Copenhagen during 1963-95. MAIN OUTCOME MEASURE: Standardised incidence ratios of testicular cancer compared with total population of Danish men. RESULTS: Men in couples with fertility problems were more likely to develop testicular cancer than other men (89 cases, standardised incidence ratio 1.6; 95% confidence interval 1.3 to 1.9). The risk was relatively constant with increasing time between semen analysis and cancer diagnosis. Analysis according to specific semen characteristics showed that low semen concentration (standardised incidence ratio 2.3), poor motility of the spermatozoa (2.5), and high proportion of morphologically abnormal spermatozoa (3.0) were all associated with an increased risk of testicular cancer. The only other cancer group that showed increased incidence was "peritoneum and other digestive organs" (six cases; 3.7, 1.3 to 8.0). Of these, two cases were probably and two cases were possibly extragonadal germ cell tumours. CONCLUSIONS: The results point towards the existence of common aetiological factors for low semen quality and testicular cancer. Low semen quality may also be associated with increased incidence of extragonadal germ cell tumours. PMID- 11009516 TI - Towards evidence based circumcision of English boys: survey of trends in practice. PMID- 11009517 TI - Wise words PMID- 11009518 TI - Prevalence of postherpetic neuralgia after a first episode of herpes zoster: prospective study with long term follow up. AB - OBJECTIVE: To estimate the frequency, duration, and clinical importance of postherpetic neuralgia after a single episode of herpes zoster. DESIGN: Prospective cohort study with long term follow up. SETTING: Primary health care in Iceland. PARTICIPANTS: 421 patients with a single episode of herpes zoster. MAIN OUTCOME MEASURES: Age and sex distribution of patients with herpes zoster, point prevalence of postherpetic neuralgia, and severity of pain at 1, 3, 6, and 12 months and up to 7.6 years after the outbreak of zoster. RESULTS: Among patients younger than 60 years, the risk of postherpetic neuralgia three months after the start of the zoster rash was 1.8% (95% confidence interval 0.59% to 4.18%) and pain was mild in all cases. In patients 60 years and older, the risk of postherpetic neuralgia increased but the pain was usually mild or moderate. After three months severe pain was recorded in two patients older than 60 years (1.7%, 2.14% to 6.15%). After 12 months no patient reported severe pain and 14 patients (3.3%) had mild or moderate pain. Seven of these became pain free within two to seven years, and five reported mild pain and one moderate pain after 7.6 years of follow up. Sex was not a predictor of postherpetic neuralgia. Possible immunomodulating comorbidity (such as malignancy, systemic steroid use, diabetes) was present in 17 patients. CONCLUSIONS: The probability of longstanding pain of clinical importance after herpes zoster is low in an unselected population of primary care patients essentially untreated with antiviral drugs. PMID- 11009519 TI - Disseminating health information in developing countries: the role of the internet. PMID- 11009520 TI - Addressing men's health issues PMID- 11009521 TI - Science, medicine, and the future: Leishmaniasis. PMID- 11009522 TI - Tulips for sale PMID- 11009523 TI - ABC of colorectal cancer: Epidemiology. PMID- 11009525 TI - Strengthening health research capacity in developing countries: a critical element for achieving health equity. PMID- 11009524 TI - Why has so little changed in maternal and child health in south Asia? PMID- 11009526 TI - Health technology transfer. PMID- 11009527 TI - Coordinating health research to promote action: the Tanzanian experience. PMID- 11009528 TI - A new look at international research ethics. PMID- 11009529 TI - The greater miracle PMID- 11009530 TI - Moving to research partnerships in developing countries. PMID- 11009532 TI - Promoting community health: from pholela to jerusalem PMID- 11009531 TI - Disaster relief and development could be twinned in health research. PMID- 11009533 TI - Knowledge can flow from developing to developed countries. PMID- 11009534 TI - Let's consider ethics of medical practice first. PMID- 11009535 TI - Information gaps have the same causes as wealth gaps. PMID- 11009537 TI - Don't forget traditional medical care. PMID- 11009536 TI - Improving access to reliable information in developing countries. PMID- 11009538 TI - Asking "how?" rather than "what, why, where, and who?". PMID- 11009539 TI - Let's start helping our neighbour. PMID- 11009540 TI - Researching on a shoestring in Colombia. PMID- 11009541 TI - Community participation is essential in clinical trials. PMID- 11009542 TI - Effect of drug patents in developing countries. PMID- 11009543 TI - Vaccines and medicines for the world's poorest. Quality of vaccines and medicines must be monitored. PMID- 11009545 TI - When public health may not be public health. PMID- 11009544 TI - Vaccines and medicines for the world's poorest. Attempts of global forum for health research should be viewed with optimistic scepticism. PMID- 11009546 TI - Accurate figures would help to assess countries' needs better. PMID- 11009548 TI - According to the charity public concern at work PMID- 11009547 TI - Maternal mortality and mothers' deaths as development indicators. PMID- 11009549 TI - The disability discrimination act requires employers to make PMID- 11009550 TI - Doctors who wish to work in developing countries should PMID- 11009551 TI - A pair of ACEs, for openers? PMID- 11009552 TI - Targeting Rho in cardiovascular disease. PMID- 11009553 TI - Na(+)-Ca(2+) exchange in failing myocardium: friend or foe? PMID- 11009554 TI - Novel genes for mitogen-independent smooth muscle replication. PMID- 11009555 TI - Tails of the L-type Ca(2+) channel: to sense oxygen or not. PMID- 11009556 TI - Splice variants reveal the region involved in oxygen sensing by recombinant human L-type Ca(2+) channels. AB - Regulation of vascular smooth muscle Ca(2+) channels by oxygen tension contributes importantly to hypoxic vasodilatation. We previously described the inhibitory effects of hypoxia on the recombinant human cardiac L-type Ca(2+) channel alpha(1C) subunit (hHT isoform) expressed in HEK 293 cells. We now demonstrate that hypoxia inhibits only one of the three naturally occurring splice variants of this channel that differ only in the C-terminal domain, permitting identification of a 71-amino acid insert in the C-terminal region of the channel that confers oxygen sensitivity. Selective restriction of the spliced insert allowed determination of a 39-amino acid region essential for oxygen sensing. This represents the first identification of the structural region of an ion channel required for sensing changes in oxygen tension. PMID- 11009557 TI - Nitric oxide activates telomerase and delays endothelial cell senescence. PMID- 11009558 TI - The preconditioning phenomenon: A tool for the scientist or a clinical reality? AB - The possibility that an innate mechanism of myocardial protection might be inducible in the human heart has generated considerable excitement and enthusiastic research. The potential to enhance myocardial resistance to ischemic injury in patients suffering the consequences of coronary artery disease has led to studies with more direct clinical relevance. However, in common with many other areas of clinical interest based on advances in basic scientific understanding, early enthusiasm may be disproportionate to ultimate therapeutic significance. There can be little doubt that our understanding of the mechanisms underlying the pathogenesis of ischemia-reperfusion injury has been enhanced significantly by the plethora of research stimulated by interest in endogenous myocardial protection. Direct extrapolation of observations in the laboratory to the cardiology clinic or operating theater is tempting but should be avoided. The results of recent clinical experiments that suggest that preconditioning can protect against ischemia, although encouraging, should be interpreted cautiously, with particular attention to the limitations of the end points available. A reasoned evaluation of recent research should prevent unrealistic expectations and allow improved design of future trials so that this potent adaptive phenomenon can be exploited to its maximum potential. PMID- 11009559 TI - Angiotensin II-induced Ca(2+) influx in renal afferent and efferent arterioles: differing roles of voltage-gated and store-operated Ca(2+) entry. AB - Angiotensin II (Ang II)-induced Ca(2+) signaling was studied in isolated rat renal arterioles using fura-2. Ang II (10 nmol/L) caused a sustained elevation in [Ca(2+)](i), which was dependent on [Ca(2+)](o) in both vessel types. This response was blocked by nifedipine in only the afferent arteriole. Using the Mn(2+) quench technique, we found that Ang II stimulates Ca(2+) influx in both vessels. Nifedipine blocked the Ang II-induced Ca(2+) influx in afferent arterioles but not in efferent arterioles. In contrast to Ang II, KCl-induced depolarization stimulated Ca(2+) influx in only the afferent arteriole. Cyclopiazonic acid (CPA, 30 micromol/L) was used to examine the presence of store operated Ca(2+) entry in myocytes isolated from each arteriole. In efferent myocytes, CPA induced a sustained Ca(2+) increase that was dependent on [Ca(2+)](o) and insensitive to nifedipine. This mechanism was absent in afferent myocytes. SKF 96365 inhibited Ang II-induced Ca(2+) entry in efferent arterioles and CPA-induced Ca(2+) entry in efferent myocytes over identical concentrations. Our findings thus indicate that Ang II activates differing Ca(2+) influx mechanisms in pre- and postglomerular arterioles. In the afferent arteriole, Ang II activates dihydropyridine-sensitive L-type Ca(2+) channels, presumably by membrane depolarization. In the efferent arteriole, Ang II appears to stimulate Ca(2+) entry via store-operated Ca(2+) influx. PMID- 11009560 TI - Early activation of the multicomponent signaling complex associated with focal adhesion kinase induced by pressure overload in the rat heart. AB - Mechanical overload elicits functional and structural adaptive mechanisms in cardiac muscle. Signaling pathways linked to integrin/cytoskeleton complexes may have a function in mediation of the effects of mechanical stimulus in myocardial cells. We investigated the tyrosine phosphorylation and the assembly of the multicomponent signaling complex associated with focal adhesion kinase (Fak) and the actin cytoskeleton in the overloaded myocardium of rats. Pressure overload induced a 3-fold increase in Fak tyrosine phosphorylation within 3 minutes after a 60-mm Hg rise in aortic pressure. A pressure stimulus that lasted for 60 minutes was accompanied by a 5-fold increase in the amount of tyrosine phosphorylated Fak, and a stimulus as low as 10 mm Hg doubled the amount of tyrosine-phosphorylated Fak in the myocardium within 10 minutes. Pressure overload also induced a time-dependent association of actin with Fak and an increase in the amount of Fak detected in the cytoskeletal fraction of the myocardium. These events were paralleled by c-Src activation and binding to Fak and by an association of Grb2 and p85 subunit of phosphatidylinositol 3-kinase with Fak. Erk1/2 and Akt, two possible downstream effectors of Fak via Grb2 and phosphatidylinositol 3-kinase, were also shown to be activated in parallel with Fak. These findings show that pressure overload induced a rapid activation of the Fak multiple signaling complex in the myocardium of rats, which suggests that this mechanism may have a role in mechanotransduction in the myocardium. PMID- 11009561 TI - Oral administration of tetrahydrobiopterin prevents endothelial dysfunction and vascular oxidative stress in the aortas of insulin-resistant rats. AB - We have reported that a deficiency of tetrahydrobiopterin (BH(4)), an active cofactor of endothelial NO synthase (eNOS), contributes to the endothelial dysfunction through reduced eNOS activity and increased superoxide anion (O(2)( )) generation in the insulin-resistant state. To further confirm this hypothesis, we investigated the effects of dietary treatment with BH(4) on endothelium dependent arterial relaxation and vascular oxidative stress in the aortas of insulin-resistant rats. Oral supplementation of BH(4) (10 mg. kg(-1). d(-1)) for 8 weeks significantly increased the BH(4) content in cardiovascular tissues of rats fed high levels of fructose (fructose-fed rats). Impairment of endothelium dependent arterial relaxation in the aortic strips of the fructose-fed rats was reversed with BH(4) treatment. The BH(4) treatment was associated with a 2-fold increase in eNOS activity as well as a 70% reduction in endothelial O(2)(-) production compared with those in fructose-fed rats. The BH(4) treatment also partially improved the insulin sensitivity and blood pressure, as well as the serum triglyceride concentration, in the fructose-fed rats. Moreover, BH(4) treatment of the fructose-fed rats markedly reduced the lipid peroxide content of both aortic and cardiac tissues and inhibited the activation of 2 redox-sensitive transcription factors, nuclear factor-kappaB and activating protein-1, which were increased in fructose-fed rats. The BH(4) treatment of control rats did not have any significant effects on these parameters. These results indicate that BH(4) augmentation is essential for the restoration of eNOS function and the reduction of vascular oxidative stress in insulin-resistant rats. PMID- 11009562 TI - Hyperglycemia inhibits vascular smooth muscle cell apoptosis through a protein kinase C-dependent pathway. AB - We hypothesized that the pathogenesis of diabetic vasculopathy involves the abnormal regulation of vascular smooth muscle cell (VSMC) apoptosis. In nondiabetic mice, a reduction in carotid artery blood flow resulted in a significant loss of medial VSMCs via apoptosis (normal flow 84+/-1 viable VSMCs, reduced flow 70+/-5 viable VSMCs; n=12, P:<0.01). In contrast, flow-induced VSMC apoptosis was markedly attenuated in streptozotocin-induced diabetic mice (normal flow 85+/-2 viable VSMC, reduced flow 82+/-4 viable VSMC; n=13, NS). In accord with our in vivo findings, the exposure of cultured rat and human VSMCs to high glucose (17.5 mmol/L) significantly attenuated the induction of apoptosis in response to serum withdrawal (rat VSMCs in normal [5.5 mmol/L] glucose 28+/-1%, high D-glucose 19+/-2%; P:<0.0001). High glucose also inhibited apoptosis induced by Fas ligand (100 ng/mL) (normal 23+/-2%, high D-glucose 13+/-2%; P:<0.006). Supplementation with the nonmetabolized enantiomer L-glucose had no effect. We confirmed reports that high glucose activates protein kinase C (PKC) and demonstrated that PKC blockade with long-term phorbol ester treatment or calphostin C prevented the antiapoptotic effect (P:<0. 001). Moreover, the upregulation of either PKCalpha or PKCbetaII expression was sufficient to inhibit serum withdrawal-induced apoptosis (control 25+/-2%, PKCalpha 11+/-2%, PKCbetaII 8+/-2%; P:<0. 0001), whereas the upregulation of PKCdelta had no significant effect. Taken together, these findings demonstrate that hyperglycemia inhibits VSMC apoptosis via a PKC-dependent pathway. PMID- 11009563 TI - Impaired contractile performance of cultured rabbit ventricular myocytes after adenoviral gene transfer of Na(+)-Ca(2+) exchanger. AB - Na(+)-Ca(2+) exchanger (NCX) gene expression is increased in the failing human heart. We investigated the hypothesis that upregulation of NCX can induce depressed contractile performance. Overexpression of NCX was achieved in isolated rabbit ventricular myocytes through adenoviral gene transfer (Ad-NCX). After 48 hours, immunoblots revealed a virus dose-dependent increase in NCX protein. Adenoviral beta-galactosidase transfection served as a control. The fractional shortening (FS) of electrically stimulated myocytes was analyzed. At 60 min(-1), FS was depressed by 15.6% in the Ad-NCX group (n=143) versus the control group (n=163, P:<0.05). Analysis of the shortening-frequency relationship showed a steady increase in FS in the control myocytes (n=26) from 0.027+/-0.002 at 30 min(-1) to 0. 037+/-0.002 at 120 min(-1) (P:<0.05 versus 30 min(-1)) and to 0. 040+/-0.002 at 180 min(-1) (P:<0.05 versus 30 min(-1)). Frequency potentiation of shortening was blunted in NCX-transfected myocytes (n=27). The FS was 0.024+/ 0.002 at 30 min(-1), 0.029+/-0.002 at 120 min(-1) (P:<0.05 versus 30 min(-1), P:<0.05 versus control), and 0. 026+/-0.002 at 180 min(-1) (NS versus 30 min(-1), P:<0.05 versus control). Caffeine contractures, which indicate sarcoplasmic reticulum Ca(2+) load, were significantly reduced at 120 min(-1) in NCX transfected cells. An analysis of postrest behavior showed a decay of FS with longer rest intervals in control cells. Rest decay was significantly higher in the Ad-NCX group; after 120 seconds of rest, FS was 78+/-4% in control and 65+/ 3% in the Ad-NCX group (P:<0.05) relative to steady-state FS before rest (100%). In conclusion, the overexpression of NCX in rabbit cardiomyocytes results in the depression of contractile function. This supports the hypothesis that upregulation of NCX can result in systolic myocardial failure. PMID- 11009564 TI - Contractile reserve and intracellular calcium regulation in mouse myocytes from normal and hypertrophied failing hearts. AB - Mouse myocyte contractility and the changes induced by pressure overload are not fully understood. We studied contractile reserve in isolated left ventricular myocytes from mice with ascending aortic stenosis (AS) during compensatory hypertrophy (4-week AS) and the later stage of early failure (7-week AS) and from control mice. Myocyte contraction and [Ca(2+)](i) transients with fluo-3 were measured simultaneously. At baseline (0.5 Hz, 1.5 mmol/L [Ca(2+)](o), 25 degrees C), the amplitude of myocyte shortening and peak-systolic [Ca(2+)](i) in 7-week AS were not different from those of controls, whereas contraction, relaxation, and the decline of [Ca(2+)](i) transients were slower. In response to the challenge of high [Ca(2+)](o), fractional cell shortening was severely depressed with reduced peak-systolic [Ca(2+)](i) in 7-week AS compared with controls. In response to rapid pacing stimulation, cell shortening and peak-systolic [Ca(2+)](i) increased in controls, but this response was depressed in 7-week AS. In contrast, the responses to both challenge with high [Ca(2+)](o) and rapid pacing in 4-week AS were similar to those of controls. Although protein levels of Na(+)-Ca(2+) exchanger were increased in both 4-week and 7-week AS, the ratio of SR Ca(2+)-ATPase to phospholamban protein levels was depressed in 7-week AS compared with controls but not in 4-week AS. This was associated with an impaired capacity to increase sarcoplasmic reticulum Ca(2+) load during high work states in 7-week AS myocytes. In hypertrophied failing mouse myocytes, depressed contractile reserve is related to an impaired augmentation of systolic [Ca(2+)](i) and SR Ca(2+) load and simulates findings in human failing myocytes. PMID- 11009565 TI - Peroxisome proliferator-activated receptor activators inhibit lipopolysaccharide induced tumor necrosis factor-alpha expression in neonatal rat cardiac myocytes. AB - Peroxisome proliferator-activated receptors (PPARs) are transcription factors belonging to the nuclear receptor superfamily. Recently, PPAR activators have been shown to inhibit the production of proinflammatory cytokines in macrophages or vascular smooth muscle cells. It has been reported that tumor necrosis factor alpha (TNF-alpha) expression is elevated in the failing heart and that TNF-alpha has a negative inotropic effect on cardiac myocytes. Therefore, we examined the effects of PPARalpha and PPARgamma activators on expression of TNF-alpha in neonatal rat cardiac myocytes. Northern blot analysis revealed expression of PPARalpha and PPARgamma mRNA in cardiac myocytes. Immunofluorescent staining demonstrated that both PPARalpha and PPARgamma were expressed in the nuclei of cells. When cardiac myocytes were transfected with PPAR responsive element (PPRE) luciferase reporter plasmid, both PPARalpha and PPARgamma activators increased the promoter activity. Cardiomyocytes were stimulated with lipopolysaccharide (LPS), and the levels of TNF-alpha in the medium were measured by ELISA. After exposure to LPS, the levels of TNF-alpha significantly increased. However, pretreatment of myocytes with PPARalpha or PPARgamma activators decreased LPS induced expression of TNF-alpha in the medium. Both PPARalpha and PPARgamma activators also inhibited LPS-induced increase in TNF-alpha mRNA in myocytes. In addition, electrophoretic mobility shift assays demonstrated that PPAR activators reduced LPS-induced nuclear factor-kappaB activation. These results suggest that both PPARalpha and PPARgamma activators inhibit cardiac expression of TNF-alpha in part by antagonizing nuclear factor-kappaB activity and that treatment with PPAR activators may lead to improvement in congestive heart failure. PMID- 11009566 TI - Angiopoietin-1 is an antipermeability and anti-inflammatory agent in vitro and targets cell junctions. AB - Inflammation is a basic pathological mechanism that underlies many diseases. An important component of the inflammatory response is the passage of plasma components and leukocytes from the blood vessel into the tissues. The endothelial monolayer lining blood vessels reacts to stimuli such as thrombin or vascular endothelial growth factor by changes in cell-cell junctions, an increase in permeability, and the leakage of plasma components into tissues. Other stimuli, such as tumor necrosis factor-alpha (TNF-alpha), are responsible for stimulating the transmigration of leukocytes. Here we show that angiopoietin-1, a cytokine essential in fetal angiogenesis, not only supports the localization of proteins such as platelet endothelial cell adhesion molecule-1 (PECAM-1) into junctions between endothelial cells and decreases the phosphorylation of PECAM-1 and vascular endothelial cadherin, but it also strengthens these junctions, as evidenced by a decrease in basal permeability and inhibition of permeability responses to thrombin and vascular endothelial growth factor. Furthermore, angiopoietin-1 inhibits TNF-alpha-stimulated leukocyte transmigration. Angiopoietin-1 may thus have a major role in maintaining the integrity of endothelial monolayers. PMID- 11009567 TI - Novel embryonic genes are preferentially expressed by autonomously replicating rat embryonic and neointimal smooth muscle cells. AB - We sought to identify and characterize the expression pattern of genes expressed by smooth muscle cells (SMCs) during periods of self-driven replication during vascular development and after vascular injury. Primary screening of a rat embryonic aortic SMC-specific cDNA library was accomplished with an autonomous embryonic SMC-enriched, nonautonomous adult SMC-subtracted cDNA probe. Positive clones were rescreened in parallel with embryonic SMC-specific and adult SMC specific cDNA probes. We identified 14 clones that hybridized only with the embryonic cDNA ("emb" clones), 11 of which did not share significant homology with sequences in any of the databases. Five of these novel emb genes (emb7, emb8, emb20, emb37, and emb41) were selectively and only transiently reexpressed in vivo by neointimal SMCs during periods of rapid replication. The emb8:embryonic growth-associated protein (EGAP), which was studied the most extensively, was expressed at high levels by cultured, autonomously replicating embryonic and neointimal SMCs but was detected only at low levels even in mitogenically stimulated adult SMCs. Finally, the administration of antisense EGAP oligonucleotides markedly attenuated embryonic and neointimal SMC replication rates. We suggest that autonomous replication of SMCs may be essential for normal vascular morphogenesis and for the vascular response to injury and that these newly identified "embryonic" genes may be part of the molecular machinery that drives this unique growth phenotype. PMID- 11009568 TI - Involvement of Rho GTPases in the transcriptional inhibition of preproendothelin 1 gene expression by simvastatin in vascular endothelial cells. AB - Endothelial dysfunction is characterized by an impaired vasodilatory response to endothelial agonists as well as by alterations in adhesion and coagulation processes. 3-Hydroxy-3-methylglutaryl-CoA reductase inhibitors (statins) have been shown to be useful in the reversal of endothelial dysfunction, an effect that may be independent of the reduction in cholesterol levels. Both the L arginine-nitric oxide-cGMP and endothelin pathways are involved in the regulation of vascular tone. Here, we show that the basal transcription rate of the preproendothelin-1 gene was decreased by simvastatin (10 micromol/L) in bovine aortic endothelial cells. Transfection studies with the preproendothelin-1 gene promoter showed that mevalonate (100 micromol/L) was able to prevent the inhibitory effect mediated by simvastatin. Protein geranylgeranylation, but not farnesylation, proved to be crucial for a correct expression of the preproendothelin-1 gene. The C3 exotoxin from Clostridium botulinum that selectively inactivates Rho GTPases, the processing of which involves geranylgeranylation, reproduced the inhibitory effect of simvastatin on the expression of preproendothelin-1. Overexpression of dominant-negative mutants of RhoA and RhoB led to a significant reduction in the preproendothelin-1 promoter activity, whereas the expression of wild-type and constitutively active forms of these proteins resulted in an increase, in support that Rho proteins are required for the basal expression of the preproendothelin-1 gene. Finally, we show that the Rho-dependent activation of the preproendothelin-1 gene transcription was inhibited by simvastatin. Thus, the control of vascular tone and proliferative response mediated by endothelin-1 is regulated at multiple levels, among which the Rho proteins play an essential role. PMID- 11009569 TI - A comparison of aorta and vena cava medial message expression by cDNA array analysis identifies a set of 68 consistently differentially expressed genes, all in aortic media. AB - We performed a systematic analysis of gene expression in arteries and veins by comparing message profiles of macaque aorta and vena cava media using a cDNA array containing 4048 known human genes, approximately 35% of currently named human genes (approximately 11,000). The data show extensive differences in RNA expression in artery versus vein media. Sixty-eight genes had consistent elevation in message expression by the aorta, but none were elevated in the vena cava. The most differentially expressed gene was regulator of G-protein signaling (RGS) 5, at an expression ratio of 46.5+/-12.6 (mean+/-SEM). The data set also contained 2 genes already known to be expressed in the aorta, elastin at 5.0+/ 1.4, and the aortic preferentially expressed gene 1 (APEG-1) at 2.3+/-0.6. We chose to analyze RGS5 expression further because of its high level of differential expression in the aorta. Levels of RGS5 mRNA were confirmed by Northern analysis and in situ hybridization. A human tissue RNA dot blot showed that RGS5 message is highest in aorta, followed by small intestine, stomach, and then heart. Northern analysis confirmed that RGS5 expression in human aorta is higher than in any region of the heart. RGS5 is a G-protein signaling regulator of unknown specificity most homologous to RGS4, an inhibitory regulator of pressure-induced cardiac hypertrophy. The expression pattern of the 68 differential genes as a whole is a start toward identifying the molecular phenotypes of arteries and veins on a systematic basis. PMID- 11009570 TI - Phosphate regulation of vascular smooth muscle cell calcification. AB - Vascular calcification is a common finding in atherosclerosis and a serious problem in diabetic and uremic patients. Because of the correlation of hyperphosphatemia and vascular calcification, the ability of extracellular inorganic phosphate levels to regulate human aortic smooth muscle cell (HSMC) culture mineralization in vitro was examined. HSMCs cultured in media containing normal physiological levels of inorganic phosphate (1.4 mmol/L) did not mineralize. In contrast, HSMCs cultured in media containing phosphate levels comparable to those seen in hyperphosphatemic individuals (>1.4 mmol/L) showed dose-dependent increases in mineral deposition. Mechanistic studies revealed that elevated phosphate treatment of HSMCs also enhanced the expression of the osteoblastic differentiation markers osteocalcin and Cbfa-1. The effects of elevated phosphate on HSMCs were mediated by a sodium-dependent phosphate cotransporter (NPC), as indicated by the ability of the specific NPC inhibitor phosphonoformic acid, to dose dependently inhibit phosphate-induced calcium deposition as well as osteocalcin and Cbfa-1 gene expression. With the use of polymerase chain reaction and Northern blot analyses, the NPC in HSMCs was identified as Pit-1 (Glvr-1), a member of the novel type III NPCs. These data suggest that elevated phosphate may directly stimulate HSMCs to undergo phenotypic changes that predispose to calcification and offer a novel explanation of the phenomenon of vascular calcification under hyperphosphatemic conditions. The full text of this article is available at http://www.circresaha.org. PMID- 11009571 TI - iNOS gene expression modulates microvascular responsiveness in endotoxin challenged mice. AB - Septic shock is characterized by vasodilation and decreased responsiveness to vasoconstrictors. Recent studies suggest this results from nitric oxide (NO) overproduction after expression of the calcium-independent isoform of NO synthase (iNOS) in smooth muscle cells. However, direct evidence linking iNOS (NOS2) expression and decreased microvascular responsiveness after septic stimuli is lacking. In the present study, we determined the effect of bacterial lipopolysaccharide (LPS, 20 mg/kg, IP) on smooth muscle contraction and endothelial relaxation in mesenteric resistance arteries from wild-type and iNOS knockout mice. Four hours after challenge with LPS or saline in vivo, concentration-dependent responses to norepinephrine (NE) and acetylcholine (NE+ACh) were measured in cannulated, pressurized vessels ex vivo. In vessels from wild-type mice, NE-induced contraction was markedly impaired after LPS, and pretreatment with the iNOS inhibitor aminoguanidine (AG) partly restored the NE contraction. In contrast, NE contraction in microvessels from iNOS knockout mice was unaffected by LPS. ACh-induced relaxation was unaffected by LPS in vessels from either genotype. These data provide direct evidence that iNOS gene expression mediates the LPS-induced decrease in microvascular responsiveness to vasoconstrictors. Moreover, the observation that AG did not fully restore NE contraction after LPS, whereas iNOS gene deficiency did, suggests that iNOS expression plays a central role in the development of the NO-independent effect of LPS on microvascular responsiveness. Finally, our data indicate that LPS or iNOS expression has little effect on endothelium-dependent relaxation, and eNOS activity does not appear to play a role in the decreased smooth muscle responsiveness after LPS in this model. The full text of this article is available at http://www.circresaha.org. PMID- 11009572 TI - Challenges in medical education-what the doctor ordered? PMID- 11009574 TI - A lay doctor's guide to the inflammatory process in the gastrointestinal tract. PMID- 11009573 TI - Pharmacological treatment of Parkinson's disease. PMID- 11009575 TI - Review of cryptogenic fibrosing alveolitis, including current treatment guidelines. PMID- 11009576 TI - Association of anticardiolipin antibodies with vascular injury: possible mechanisms. PMID- 11009578 TI - Emptiness of the left iliac fossa: a new clinical sign of sigmoid volvulus. AB - BACKGROUND: Diagnostic difficulties are common in sigmoid volvulus. This diagnostic delay contributes to high morbidity and mortality. AIM: This paper recognises visible or palpable emptiness of the left iliac fossa as a new clinical sign of sigmoid volvulus. Predictive accuracy of the sign was assessed in this study. METHODS: 519 patients with acute abdomen of non-traumatic origin were prospectively studied. RESULTS: The new sign was present in 24 out of 86 patients (28%) with sigmoid volvulus. It was absent in all those who did not have sigmoid volvulus. The positive predictive value of the sign was 100%. CONCLUSION: Emptiness of the left iliac fossa is a valuable diagnostic adjunct. Positive sign is pathognomonic of sigmoid volvulus. Nevertheless, a negative sign warrants further investigations. PMID- 11009579 TI - Single photon emission computed tomography in tuberculous meningitis. AB - INTRODUCTION: Data on single photon emission computed tomography (SPECT) in tuberculous meningitis are lacking and prompted this study. SPECT findings in tuberculous meningitis are reported and correlated with clinical and radiological findings. PATIENTS AND METHODS: Seventeen patients with tuberculous meningitis that was diagnosed on clinical, radiological, and laboratory criteria have been included. Their age ranged between 5 and 62 years and four of them were female. Computed tomography and/or magnetic resonance imaging (MRI) and SPECT using (99m)Tc-ethylene cystine dimer were performed in all the patients. On the basis of Barthel index (BI) score the patients' outcome was defined as complete (BI = 20), partial (BI = 19-12), and poor recovery (BI<12). RESULT: Eleven patients were in stage III and three each in stage II and stage I tuberculous meningitis. Two patients had hemiplegia and five quadriplegia. Computed tomography was abnormal in 11 out of 16 patients and revealed hydrocephalus in nine, basal exudates, infarction in subcortical white matter and basal ganglia in six patients each, frontal cortical infarction in one, and granulomata in three patients. Cranial MRI was carried out in four patients and revealed multiple granulomata, hydrocephalus, and brainstem infarction in two patients each. SPECT studies were abnormal in all except two patients and revealed hypoperfusion of the basal ganglia in 14, cortical hypoperfusion in 10, and midbrain hypoperfusion in one patient. At the three month follow up four patients had died, five had poor, three partial, and five complete recovery. The SPECT studies were more frequently abnormal compared with computed tomography but did not correlate with stage of meningitis or outcome. CONCLUSION: In tuberculous meningitis subcortical and cortical hypoperfusion is common but it does not correlate with stage of meningitis or three month outcome. PMID- 11009577 TI - Behcet's disease. AB - Behcet's disease is a systemic vasculitis of unknown aetiology characteristically affecting venules. Onset is typically in young adults with recurrent oral and genital ulceration, uveitis, skin manifestations, arthritis, neurological involvement, and a tendency to thrombosis. It has a worldwide distribution but is prevalent in Japan, the Middle East, and some Mediterranean countries. International diagnostic criteria have been proposed, however diagnosis can be problematical, particularly if the typical ulcers are not obvious at presentation. Treatment is challenging, must be tailored to the pattern of organ involvement for each patient and often requires combination therapies. PMID- 11009580 TI - Closed pelvic fractures: characteristics and outcomes in older patients admitted to medical and geriatric wards. AB - OBJECTIVE: To investigate the characteristics and outcomes of older patients with pelvic fracture admitted to medical and geriatric wards. METHODS: All patients admitted to medical and geriatric wards with a pelvic fracture over a four year period were identified using the hospital clinical coding database. Data were collected from casenotes, hospital and Family Health Services Authority databases. Where available, pelvic radiographs were graded according to the Singh index. RESULTS: The casenotes of 148 patients (126 women) were studied; 83% (n=123) of patients suffered a pelvic fracture in low energy trauma. Mean (SD) length of hospital stay was 21.3 (17.6) days. Single breaks of the pubic rami accounted for 47.2% (n=68) of all fractures. Inpatient mortality was 7.6% and at one year was 27%. There was a marked adverse effect on the mobility of survivors with all patients using at least a walking stick at discharge and 51.1% (n=70) needing assistance for mobility. Although 70.9% (n=83) of patients admitted from home (or warden aided accommodation) were able to return there, 84.3% (n=70) of them required extra community support. Rates of institutionalisation rose from 20.9% (n=31) at admission to 35.8% (49/137) of survivors at discharge. Altogether 93% (n=107) of 115 patients, in whom adequate quality pelvic radiographs were available, were assigned a Singh index grade of 4 or less indicating the presence of osteoporosis. CONCLUSIONS: Pelvic fractures are often the result of low energy trauma. They are associated with appreciable inpatient and considerable one year mortality. They also have marked negative effects on mobility in the short term. They result in increased levels of dependency in terms of higher levels of community support and rates of institutionalisation. On the evidence of Singh index grading, pelvic fractures are associated with low bone density. PMID- 11009581 TI - QT prolongation due to roxithromycin. PMID- 11009582 TI - Commentary-QT prolongation due to roxithromycin. PMID- 11009583 TI - An unusual case of pyrexia of unknown origin with cervical lymphadenopathy. AB - Kikuchi's disease is usually a self limiting illness characterised by pyrexia, neutropenia, and cervical lymphadenopathy particularly in young women of Asian descent. This often leads to an initial misdiagnosis of lymphoma. A case of a young Asian woman who presented with pyrexia of unknown origin is described. PMID- 11009584 TI - Carotid sinus syndrome masquerading as treatment resistant epilepsy. AB - A 65 year old woman had a 12 year history of frequent, recurrent seizure-like episodes labelled as treatment resistant epilepsy after neurological evaluation and follow up and treatment with multiple antiepileptic medications. Carotid sinus massage provoked 5.6 seconds asystole with symptom reproduction, and she has remained symptom-free after permanent pacemaker implantation for her carotid sinus syndrome and withdrawal of antiepileptic medications. PMID- 11009585 TI - Chorea disclosing deterioration of polycythaemia vera. AB - Neurological manifestations occur frequently in polycythaemia. Chorea, however, is a rare complication of the disease. A case of chorea in a patient previously diagnosed with polycythaemia vera is reported. Choreic movements started after measurement of haematological variables showed deterioration. It was considered that this was caused by inappropriate treatment with iron because the chorea was markedly reduced after the two first venesections and normalisation of the packed cell volume and haemoglobin parameters. PMID- 11009586 TI - A case of migratory lymphadenopathy and cutaneous anergy in an Asian woman. PMID- 11009587 TI - Multiple focal lesions in liver and spleen in acute leukaemia PMID- 11009588 TI - A pregnant patient with bilateral ischaemic limbs. PMID- 11009589 TI - Severe symptomatic hypercalcaemia. PMID- 11009590 TI - A case of migratory lymphadenopathy and cutaneous anergy in an asian woman PMID- 11009591 TI - Multiple focal lesions in liver and spleen in acute leukaemia. PMID- 11009592 TI - A pregnant patient with bilateral ischaemic limbs PMID- 11009593 TI - Severe symptomatic hypercalcaemia PMID- 11009594 TI - Cultural sensitivity. PMID- 11009595 TI - The functional association of polymyxin B with bacterial lipopolysaccharide is stereospecific: studies on polymyxin B nonapeptide. AB - The Gram-negative bacterial endotoxin lipopolysaccharide (LPS) is a major inducer of sepsis. The natural cyclic peptide polymyxin B (PMB) is a potent antimicrobial agent, albeit highly toxic, by virtue of its capacity to neutralize the devastating effects of LPS. However, the exact mode of association between PMB and LPS is not clear. In this study, we have synthesized polymyxin B nonapeptide, the LPS-binding cyclic domain of PMB, and its enantiomeric analogue and studied several parameters related to their interaction with LPS and their capacity to sensitize Gram-negative bacteria toward hydrophobic antibiotics. The results suggest that whereas the binding of the two enantiomeric peptides to E. coli and to E. coli LPS is rather similar, functional association with the bacterial cell is stereospecific. Thus, the L-enantiomer is capable of synergism with the hydrophobic antimicrobial drugs novobiocin and erythromycin, whereas the D enantiomer is devoid of such activity. The potential of understanding and consequently utilizing the PMB-LPS association for novel, nontoxic PMB-derived drugs is discussed. PMID- 11009596 TI - Biochemical and biophysical characterization of OmpG: A monomeric porin. AB - A recombinant form of the porin OmpG, OmpGm, lacking the signal sequence, has been expressed in Escherichia coli. After purification under denaturing conditions, the protein was refolded in the detergent Genapol X-080, where it gained a structure rich in beta sheet as evidenced by a CD spectrum similar to that of the native form. Electrophoretic analysis and limited proteolysis experiments suggested that refolded OmpGm exists in at least three forms. Nevertheless, the recombinant protein formed uniform channels in planar bilayers with a conductance of 0.81 nS (1 M NaCl, pH 7.5). Previous biochemical studies had suggested that OmpG is a monomeric porin, rather than the usual trimer. Bilayer recordings substantiated this proposal; voltage-induced closures occurred consistently in a single step, and channel block by Gd(3+) lacked the cooperativity seen with the trimeric porin OmpF. The availability of milligram amounts of a monomeric porin will be useful both for basic studies of porin function and for membrane protein engineering. PMID- 11009597 TI - Role of the hinge region and the tryptophan residue in the synthetic antimicrobial peptides, cecropin A(1-8)-magainin 2(1-12) and its analogues, on their antibiotic activities and structures. AB - A 20-residue hybrid peptide CA(1-8)-MA(1-12) (CA-MA), incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA), has potent antimicrobial activity without toxicity against human erythrocytes. To investigate the effects of the Gly-Ile-Gly hinge sequence of CA-MA on the antibacterial and antitumor activities, two analogues in which the Gly-Ile-Gly sequence of CA-MA is either deleted (P1) or substituted with Pro (P2) were synthesized. The role of the tryptophan residue at position 2 of CA-MA on its antibiotic activity was also investigated using two analogues, in which the Trp2 residue of CA-MA is replaced with either Ala (P3) or Leu (P4). The tertiary structures of CA-MA, P2, and P4 in DPC micelles, as determined by NMR spectroscopy, have a short amphiphilic helix in the N-terminus and about three turns of alpha-helix in the C-terminus, with the flexible hinge region between them. The P1 analogue has an alpha-helix from Leu4 to Ala14 without any hinge structure. P1 has significantly decreased lytic activities against bacterial and tumor cells and PC/PS vesicles (3:1, w/w), and reduced pore-forming activity on lipid bilayers, while P2 retained effective lytic activities and pore-forming activity. The N-terminal region of P3 has a flexible structure without any specific secondary structure. The P3 modification caused a drastic decrease in the antibiotic activities, whereas P4, with the hydrophobic Leu side chain at position 2, retained its activities. On the basis of the tertiary structures, antibiotic activities, vesicle-disrupting activities, and pore-forming activities, the structure-function relationships can be summarized as follows. The partial insertion of the Trp2 of CA-MA into the membrane, as well as the electrostatic interactions between the positively charged Lys residues at the N-terminus of the CA-MA and the anionic phospholipid headgroups, leads to the primary binding to the cell membrane. Then, the flexibility or bending potential induced by the Gly-Ile-Gly hinge sequence or the Pro residue in the central part of the peptides may allow the alpha-helix in the C-terminus to span the lipid bilayer. These structural features are crucial for the potent antibiotic activities of CA-MA. PMID- 11009598 TI - Escherichia coli uracil DNA glycosylase: NMR characterization of the short hydrogen bond from His187 to uracil O2. AB - Uracil DNA glycosylase (UDG) cleaves the glycosidic bond of deoxyuridine in DNA using a hydrolytic mechanism, with an overall catalytic rate enhancement of 10(12)-fold over the solution reaction. The nature of the enzyme-substrate interactions that lead to this large rate enhancement are key to understanding enzymatic DNA repair. Using (1)H and heteronuclear NMR spectroscopy, we have characterized one such interaction in the ternary product complex of Escherichia coli UDG, the short (2.7 A) H bond between His187 N(epsilon)(2) and uracil O2. The H bond proton is highly deshielded at 15.6 ppm, indicating a short N-O distance and exhibits a solvent exchange rate that is 400- and 10(5)-fold slower than free imidazole at pH 7.5 and pH 10, respectively. Heteronuclear NMR experiments at neutral pH show that this H bond involves the neutral imidazole form of His187 and the N1-O2 imidate form of uracil. The excellent correspondence of the pK(a) for the disappearance of the H bond (pK(a) = 6.3 +/- 0.1) with the previously determined pK(a) = 6.4 for the N1 proton of enzyme-bound uracil indicates that the H bond requires negative charge on uracil O2 [Drohat, A. C., and Stivers, J. T. (2000) J. Am. Chem. Soc. 122, 1840-1841]. Although the above characteristics suggest a short strong H bond, the D/H fractionation factor of phi = 1.0 is more typical of a normal H bond. This unexpected observation may reflect a large donor-acceptor pK(a) mismatch or the net result of two opposing effects on vibrational frequencies: decreased N-H bond stretching frequencies (phi < 1) and increased bending frequencies (phi > 1) relative to the O-H bonds of water. The role of this H bond in catalysis by UDG and several approaches to quantify the H bond energy are discussed. PMID- 11009599 TI - Thermodynamic basis of resistance to HIV-1 protease inhibition: calorimetric analysis of the V82F/I84V active site resistant mutant. AB - One of the most serious side effects associated with the therapy of HIV-1 infection is the appearance of viral strains that exhibit resistance to protease inhibitors. The active site mutant V82F/I84V has been shown to lower the binding affinity of protease inhibitors in clinical use. To identify the origin of this effect, we have investigated the binding thermodynamics of the protease inhibitors indinavir, ritonavir, saquinavir, and nelfinavir to the wild-type HIV 1 protease and to the V82F/I84V resistant mutant. The main driving force for the binding of all four inhibitors is a large positive entropy change originating from the burial of a significant hydrophobic surface upon binding. At 25 degrees C, the binding enthalpy is unfavorable for all inhibitors except ritonavir, for which it is slightly favorable (-2.3 kcal/mol). Since the inhibitors are preshaped to the geometry of the binding site, their conformational entropy loss upon binding is small, a property that contributes to their high binding affinity. The V82F/I84V active site mutation lowers the affinity of the inhibitors by making the binding enthalpy more positive and making the entropy change slightly less favorable. The effect on the enthalpy change is, however, the major one. The predominantly enthalpic effect of the V82F/I84V mutation is consistent with the idea that the introduction of the bulkier Phe side chain at position 82 and the Val side chain at position 84 distort the binding site and weaken van der Waals and other favorable interactions with inhibitors preshaped to the wild-type binding site. Another contribution of the V82F/I84V to binding affinity originates from an increase in the energy penalty associated with the conformational change of the protease upon binding. The V82F/I84V mutant is structurally more stable than the wild-type protease by about 1.4 kcal/mol. This effect, however, affects equally the binding affinity of substrate and inhibitors. PMID- 11009600 TI - Formation of a noncovalent serpin-proteinase complex involves no conformational change in the serpin. Use of 1H-15N HSQC NMR as a sensitive nonperturbing monitor of conformation. AB - A structural understanding of the nature and scope of serpin inhibition mechanisms has been limited by the inability so far to crystallize any serpin proteinase complex. We describe here the application of [(1)H-(15)N]-HSQC NMR on uniformly and residue-selectively (15)N-labeled serpin alpha(1)-proteinase inhibitor (Pittsburgh variant with stabilizing mutations) to provide a nonperturbing and exquisitely sensitive means of probing the conformation of the serpin alone and in a noncovalent complex with inactive, serine 195-modified, bovine trypsin. The latter should be a good model both for the few examples of reversible serpin-proteinase complexes and for the initial Michaelis-like complex formed en route to irreversible covalent inhibition. Cleavage of the reactive center loop, with subsequent insertion into beta-sheet A, caused dramatic perturbation of most of the NMR cross-peaks. This was true for both the uniformly labeled and alanine-specifically labeled samples. The spectra of uniformly or leucine- or alanine-specifically labeled alpha(1)-proteinase inhibitor in noncovalent complex with unlabeled inactive trypsin gave almost no detectable chemical shift changes of cross-peaks, but some general increase in line width. Residue-specific assignments of the four alanines in the reactive center loop, at P12, P11, P9, and P4, allowed specific examination of the behavior of the reactive center loop. All four alanines showed higher mobility than the body of the serpin, consistent with a flexible reactive center loop, which remained flexible even in the noncovalent complex with proteinase. The three alanines near the hinge point for insertion showed almost no chemical shift perturbation upon noncovalent complex formation, while the alanine at P4 was perturbed, presumably by interaction with the active site of bound trypsin. Reporters from both the body of the serpin and the reactive center loop therefore indicate that noncovalent complex formation involves no conformational change in the body of the serpin and only minor perturbation of the reactive center loop in the region which contacts proteinase. Thus, despite the large size of serpin and serpin proteinase complex, 45 and 69 kDa respectively, NMR provides a very sensitive means of probing serpin conformation and mobility, which should be applicable both to noncovalent and to covalent complexes with a range of different proteinases, and probably to other serpins. PMID- 11009601 TI - Structural effects of protein lipidation as revealed by LysB29-myristoyl, des(B30) insulin. AB - Intracellular proteins are frequently modified by covalent addition of lipid moieties such as myristate. Although a functional role of protein lipidation is implicated in diverse biological processes, only a few examples exist where the structural basis for the phenomena is known. We employ the insulin molecule as a model to evaluate the detailed structural effects induced by myristoylation. Several lines of investigation are used to characterize the solution properties of Lys(B29)(N(epsilon)-myristoyl) des(B30) insulin. The structure of the polypeptide chains remains essentially unchanged by the modification. However, the flexible positions taken up by the hydrocarbon chain selectively modify key structural properties. In the insulin monomer, the myristoyl moiety binds in the dimer interface and modulates protein-protein recognition events involved in insulin dimer formation and receptor binding. Myristoylation also contributes stability expressed as an 30% increase in the free energy of unfolding of the protein. Addition of two Zn(2+)/hexamer and phenol results in the displacement of the myristoyl moiety from the dimer interface and formation of stable R(6) hexamers similar to those formed by human insulin. However, in its new position on the surface of the hexamer, the fatty acid chain affects the equilibria of the phenol-induced interconversions between the T(6), T(3)R(3), and R(6) allosteric states of the insulin hexamer. We conclude that insulin is an attractive model system for analyzing the diverse structural effects induced by lipidation of a compact globular protein. PMID- 11009603 TI - Comparison of the membrane interaction and permeabilization by the designed peptide Ac-MB21-NH2 and truncated dermaseptin S3. AB - Ac-MB21-NH(2) (Ac-FASLLGKALKALAKQ-NH(2)) and dermaseptin S3(1-16)-NH(2) (ALWKNMLKGIGKLAGK-NH(2)) are cationic amphipathic peptides with antimicrobial activity against a broad spectrum of microorganisms including various fungi. The interaction of the peptides with liposomes was studied by exploiting the tryptophan fluorescence of F1W-Ac-MB21-NH(2) and dermaseptin S3(1-16)-NH(2). Spectral analysis and the use of quenchers indicate that the tryptophans of both peptides insert more deeply in anionic than in zwitterionic liposomes. Membrane insertion correlates with the formation of an alpha-helical peptide structure. Both peptides permeabilize liposomes composed of anionic, cylindric phospholipids more efficiently than liposomes formed of zwitterionic, conic (phospho)lipids. PMID- 11009602 TI - Drug binding sites on P-glycoprotein are altered by ATP binding prior to nucleotide hydrolysis. AB - P-glycoprotein (P-gp) confers multiple drug resistance on cancer cells by acting as a plasma membrane localized ATP-dependent drug efflux pump. Currently, there is little information on the nature of the communication between the energy providing nucleotide binding domains (NBDs) and the drug binding sites of P-gp to generate transport of substrate. Many substrates and modulators cause alterations in ATP hydrolysis, but what effect do the various stages of the catalytic cycle have on drug interaction with P-gp? Vanadate trapping of Mg.ADP caused a reversible decrease in the binding capacity of the transported substrate [(3)H] vinblastine and the nontransported modulator [(3)H]XR9576 to P-gp in CH(r)B30 cell membranes. The non-hydrolyzable nucleotide analogue ATP-gamma-S also caused a reduction in the binding capacity of [(3)H]-vinblastine but not for the modulator [(3)H]XR9576. This indicates that signaling to the NBDs following binding of a nontransported modulator is different to that transmitted upon interaction of a transported substrate. Second, it appears that the binding of nucleotide, rather than its hydrolysis, causes the initial conformational shift in the drug-binding site during a transport cycle. PMID- 11009604 TI - Actomyosin regulatory properties of yeast tropomyosin are dependent upon N terminal modification. AB - The yeast tropomyosin 1 gene (TPM1) encodes the major isoform of the two tropomyosins (Tm) found in yeast. The gene has been expressed in E. coli and the protein purified. The gene product (yTm1) is a 199-amino acid protein that has a low affinity for actin compared to the native yTm1 purified from yeast. Mass spectrometry shows that the native protein is acetylated while the recombinant protein is not. A series of yTm1 N-terminal constructs were made with either an Ala-Ser dipeptide extension previously shown to restore actin binding to skeletal muscle Tm or the natural extension found in fibroblast Tm 5a/b. All constructs bound actin tightly and showed similar CD spectra and thermal stability. All constructs induced cooperativity in the equilibrium binding of myosin subfragment 1, to actin but the binding curves differed significantly between the constructs. The apparent cooperative unit size (n) and closed/open equilibrium (K(T)) were determined using a fluorescence titration technique [Maytum et al. (1998) Biophys. J. 74, A347]. The data could be accounted for by changes in K(T) (0.1-1) with no change in n. Values of n were approximately twice the structural unit size (5 actin sites). The presence of yTm on actin had little effect upon the overall affinity of S1 for actin despite showing an ability to regulate the acto myosin interaction. These results show that the short yTm can aid our understanding of actomyosin regulation and that the N-terminus of Tm has a major influence upon its regulatory properties. PMID- 11009605 TI - Investigation of the role of glutamine-471 and glutamine-1114 in the two catalytic sites of P-glycoprotein. AB - P-glycoprotein, also known as multidrug resistance protein, pumps drugs out of cells using ATP hydrolysis as the energy source. Glutamine-471 and the corresponding glutamine-1114 in the two catalytic sites of P-glycoprotein are conserved in ABC transporters. X-ray structures show that they lie close to the bound nucleotide. Proposed functional roles are (1) activation of the attacking water for ATP hydrolysis, (2) coordination of the essential Mg(2+) cofactor in Mg nucleotide, and (3) signal communication between catalytic site reaction chemistry and drug-binding sites. We made mutations Q471A, Q471E, Q1114A, and Q1114E in mouse MDR3 P-glycoprotein. Pure mutant and wild-type proteins were prepared and subjected to enzymatic and biochemical characterization. We conclude from the results that the primary role of this glutamine residue is in interdomain signal communication. Coordination of the Mg(2+) cofactor is not a critical functional role, neither is activation of the attacking water molecule, although an auxiliary role in positioning the water cannot be ruled out. We found that equivalent mutations (Ala or Glu) in either of the two P-glycoprotein catalytic sites produced the same effects, implying functional symmetry of the two sites. PMID- 11009606 TI - (R)-3-hydroxybutyrate dehydrogenase: selective phosphatidylcholine binding by the C-terminal domain. AB - (R)-3-Hydroxybutyrate dehydrogenase (BDH) is a lipid-requiring mitochondrial enzyme that has a specific requirement of phosphatidylcholine (PC) for function. The C-terminal domain (CTBDH) of human heart BDH (residues 195-297) has now been expressed in Escherichia coli as a chimera with a soluble protein, glutathione S transferase (GST), yielding GST-CTBDH, a novel fusion protein that has been purified and shown to selectively bind to PC vesicles. Both recombinant human heart BDH (HH-Histag-BDH) and GST-CTBDH (but not GST) form well-defined protein lipid complexes with either PC or phosphatidylethanolamine (PE)/diphosphatidylglycerol (DPG) vesicles (but not with digalactosyl diglyceride vesicles) as demonstrated by flotation in sucrose gradients. The protein-PC complexes are stable to 0.5 M NaCl, but complexes of either HH-Histag-BDH or GST CTBDH with PE/DPG vesicles are dissociated by salt treatment. Thrombin cleavage of GST-CTBDH, either before or after reconstitution with PC vesicles, yields CTBDH (12 111 Da by MALDI mass spectrometry) which retains lipid binding without attached GST. The BDH activator, 1-palmitoyl-2-(1-pyrenyl)decanoyl-PC (pyrenyl PC), at <2.5% of total phospholipid in vesicles, efficiently quenches a fraction (0.36 and 0.47, respectively) of the tryptophan fluorescence of both HH-Histag BDH and GST-CTBDH with effective Stern-Volmer quenching constants, (K(Q))(eff), of 11 and 9.3 (%)(-)(1), respectively (half-maximal quenching at approximately 0.1% pyrenyl-PC). Maximal quenching by pyrenyl-PC obtains at approximately stoichiometric pyrenyl-PC to protein ratios, reflecting high-affinity interaction of pyrenyl-PC with both HH-Histag-BDH and GST-CTBDH. The analogous pyrenyl-PE effects a similar maximal quenching of tryptophan fluorescence for both proteins but with approximately 15-fold lower (K(Q))(eff) (half-maximal quenching at approximately 1.5% pyrenyl-PE) referable to nonspecific interaction of pyrenyl-PE with HH-Histag-BDH or GST-CTBDH. Thus, the 103-residue CTBDH constitutes a PC selective lipid binding domain of the PC-requiring BDH. PMID- 11009607 TI - Fluoride effects along the reaction pathway of pyrophosphatase: evidence for a second enzyme.pyrophosphate intermediate. AB - The fluoride ion is a potent and specific inhibitor of cytoplasmic pyrophosphatase (PPase). Fluoride action on yeast PPase during PP(i) hydrolysis involves rapid and slow phases, the latter being only slowly reversible [Smirnova, I. N., and Baykov, A. A. (1983) Biokhimiya 48, 1643-1653]. A similar behavior is observed during yeast PPase catalyzed PP(i) synthesis. The amount of enzyme.PP(i) complex formed from solution P(i) exhibits a rapid drop upon addition of fluoride, followed, at pH 7.2, by a slow increase to nearly 100% of the total enzyme. The slow reaction results in enzyme inactivation, which is not immediately reversed by dilution. These data show that fluoride binds to an enzyme.PP(i) intermediate during the slow phase and to an enzyme.P(i) intermediate during the rapid phase of the inhibition. In Escherichia coli PPase, the enzyme.PP(i) intermediate binds F(-) rapidly, explaining the lack of time dependence in the inhibition of this enzyme. The enzyme.PP(i) intermediate formed during PP(i) hydrolysis binds fluoride much faster (yeast PPase) or tighter (E. coli PPase) than the similar complex existing at equilibrium with P(i). It is concluded that PPase catalysis involves two enzyme.PP(i) intermediates, of which only one (immediately following PP(i) addition and predominating at acidic pH) can bind fluoride. Simulation experiments have indicated that interconversion of the enzyme.PP(i) intermediates is a partially rate-limiting step in the direction of hydrolysis and an exclusively rate-limiting step in the direction of synthesis. PMID- 11009608 TI - Characterization of the regiochemistry and cryptoregiochemistry of a Caenorhabditis elegans fatty acid desaturase (FAT-1) expressed in Saccharomyces cerevisiae. AB - To characterize the fatty acid desaturase produced by the fat-1 gene from the nematode Caenorhabditis elegans, the functional expression of this enzyme was effected in the yeast Saccharomyces cerevisiae. The GC-MS analysis of desaturated products derived from various fatty acids, including deuterium-labeled thia fatty acids supplied to growing cultures of transformed yeast, has defined the substrate requirements, regiochemistry, and cryptoregiochemistry of the enzyme. The desaturase acts on substrates of 16-20 carbons with a preference for omega-6 fatty acids, and its regioselectivity was confirmed to be that of an omega-3 desaturase. (omega-x refers to a double bond or desaturation between carbons x and x+1, counting from the methyl end of a fatty acid.) The primary deuterium kinetic isotope effects (KIEs) at C-15 and C-16 of a C18 fatty acid analogue were measured via competitive incubation experiments: While k(H)/k(D) at the omega-3 position was shown to be large (7.8 +/- 0.4), essentially no KIE at the omega-2 position was observed (k(H)/k(D) = 0.99 +/- 0.04). This result indicates that omega-3 desaturation is initiated by an energetically difficult C-H bond cleavage at the carbon closer to the carboxyl terminus. The results are discussed in the context of a general model relating the structure and function of membrane-bound fatty acid desaturases featuring differing regioselectivities. PMID- 11009609 TI - Lysine 199 is the general acid in the NAD-malic enzyme reaction. AB - Site-directed mutagenesis was used to change K199 in the Ascaris suum NAD-malic enzyme to A and R and Y126 to F. The K199A mutant enzyme gives a 10(5)-fold decrease in V and a 10(6)-fold decrease in V/K(malate) compared to the WT enzyme. In addition, the ratio for partitioning of the oxalacetate intermediate toward pyruvate and malate changes from a value of 0.4 for the WT enzyme to 1.6 for K199A, and repeating the experiment with A-side NADD gives isotope effects of 3 and 1 for the WT and K199A mutant enzymes, respectively. The K199R mutant enzyme gives only a factor of 10 decrease in V, and the pK for the general acid in this mutant enzyme has increased from 9 for the WT enzyme to >10 for the K199R mutant enzyme. Tritium exchange from solvent into pyruvate is catalyzed by the WT enzyme, but not by the K199A mutant enzyme. The Y126F mutant enzyme gives a 10(3) fold decrease in V. The oxalacetate partition ratio and isotope effect on oxalacetate reduction for the Y126F mutant enzyme are identical, within error, to those measured for the WT enzyme. Thus, Y126 is important to the overall reaction, but its role at present is unclear. Data are consistent with K199 functioning as the general acid that protonates C3 of enolpyruvate to generate the pyruvate product in the malic enzyme reaction. PMID- 11009610 TI - Kinetic mechanism of human histone acetyltransferase P/CAF. AB - Human transcriptional coactivator P/CAF (p300/CBP-associating factor) is a histone acetyltransferase (HAT) and is a member of the GNAT (GCN5 related N acetyltransferases) superfamily. P/CAF was originally identified by its ability to activate transcription of a variety of genes through its interaction with p300/CBP. Though Lys-14 of histone H3 appears to be the preferred substrate, other nonhistone proteins can also serve as substrates for P/CAF. However, few studies have addressed the catalytic and kinetic mechanisms of histone/protein acetylation by P/CAF. In this study, we have systematically determined the kinetic mechanism for P/CAF, identified the critical ionizations for binding/catalysis, and established the rate-limiting step in turnover. This was accomplished by a variety of approaches including pH-dependent activity measurements, Bi-substrate kinetic analysis, authentic product inhibition by coenzyme A (CoA) and acetylated H3 (Ac-Lys-14) peptide, direct measurements of substrate/product binding affinities (equilibrium dialysis), and a pre-steady state quench-flow analysis. The results are consistent with a fully ordered Bi-Bi kinetic mechanism, where chemical catalysis is rate-determining. Acetyl-CoA (AcCoA) binds with high affinity (K(d) = 0.64 +/- 0.12 microM) to the free form of the enzyme. Histone H3 peptide binds (apparent K(d) = 116 +/- 17 microM) only after AcCoA is bound. No H3 peptide binding to the free enzyme was detectable. In the ternary complex, the epsilon-amino of Lys-14 (H3 peptide substrate) directly attacks the carbonyl carbon of AcCoA, transferring the acetyl group to the acceptor peptide substrate (rate-limiting step). Products are released in an ordered fashion, with Ac-Lys-14 H3 released first followed by release of CoA. The pH dependency of the k(cat)/K(m) parameter revealed two P/CAF ionizable groups (pK(a) values of 6.9 and 7.5) that must be unprotonated for activity. The group with a pK(a) value 7.5 was assigned to Glu-570, which is the proposed general base catalyst, abstracting a proton from the epsilon-amino group and facilitating nucleophilic attack. PMID- 11009611 TI - 5' --> 3' molecular polarity of human replication protein A (hRPA) binding to pseudo-origin DNA substrates. AB - Human replication protein A (hRPA) was previously seen to efficiently bind a 48 bp simian virus 40 (SV40) "pseudo-origin" (PO) substrate that mimics a DNA structure found within the SV40 T antigen-origin (ori) complex. To understand the role of hRPA during the initiation of replication, we examined the PO sequence and structure requirements for hRPA interaction. Binding and unwinding were found to be most efficient when both strands of the central 8 nt single-stranded DNA (ssDNA) bubble region contained a polypyrimidine structure, with these activities proportionately reduced when the bubble region was replaced with a purine tract on one or both strands. Examination of the importance of the two duplex flanks indicates that the early gene side contains a DNA structural feature located one duplex turn from the bubble whose mutation significantly affects the affinity of hRPA for the substrate. When present in the context of ori, mutation of this sequence was seen to have significant effects on SV40 DNA replication in vitro and on the denaturation of ori, indicating that origin activity can be modulated by cis-acting elements which alter the hRPA binding affinity. Use of fork and overhang substrates containing 8 nt pyrimidine or purine arms demonstrates that hRPA binding to DNA involves a particular molecular polarity in which initial hRPA binding occurs on the 5' side of a ssDNA substrate, and then extends in the 3' direction to create a stably bound hRPA. These data have implications on the mechanism of the initiation of eukaryotic DNA replication as well as on the sites of nascent strand synthesis within the origin. PMID- 11009612 TI - Cleavage of symmetric immobile DNA junctions by Escherichia coli RuvC. AB - The Holliday junction is a key DNA intermediate in the process of genetic recombination. It consists of two double-helical domains composed of homologous strands that flank a branch point; two of the strands are roughly helical, and two form the crossover between the helices. RuvC is a Holliday junction resolvase that cleaves the helical strands at a symmetric sequence, leading to the production of two recombinant molecules. We have determined the position of the cleavage site relative to the crossover point by the use of symmetric immobile junctions; these are DNA molecules containing two crossover points, one held immobile by sequence asymmetry and the second a symmetric sequence, but held immobile by torsional coupling to the first junction. We have built five symmetric immobile junctions, in which the tetranucleotide recognition site is moved stepwise relative to the branch point. We have used kinetic analysis of catalysis, gel retardation, and hydroxyl radical hypersensitivity to analyze this system. We conclude that the internucleotide linkage one position 3' to the crossover point is the favored site of cleavage. PMID- 11009613 TI - Interaction of the Escherichia coli replication terminator protein (Tus) with DNA: a model derived from DNA-binding studies of mutant proteins by surface plasmon resonance. AB - The Escherichia coli replication terminator protein (Tus) binds tightly and specifically to termination sites such as TerB in order to halt DNA replication. To better understand the process of Tus-TerB interaction, an assay based on surface plasmon resonance was developed to allow the determination of the equilibrium dissociation constant of the complex (K(D)) and association and dissocation rate constants for the interaction between Tus and various DNA sequences, including TerB, single-stranded DNA, and two nonspecific sequences that had no relationship to TerB. The effects of factors such as the KCl concentration, the orientation and length of the DNA, and the presence of a single-stranded tail on the binding were also examined. The K(D) measured for the binding of wild type and His(6)-Tus to TerB was 0.5 nM in 250 mM KCl. Four variants of Tus containing single-residue mutations were assayed for binding to TerB and the nonspecific sequences. Three of these substitutions (K89A, R198A, and Q250A) increased K(D) by 200-300-fold, whereas the A173T substitution increased K(D) by 4000-fold. Only the R198A substitution had a significant effect on binding to the nonspecific sequences. The kinetic and thermodynamic data suggest a model for Tus binding to TerB which involves an ordered series of events that include structural changes in the protein. PMID- 11009614 TI - Amphipathic helices support function of blood coagulation factor IXa. AB - Blood coagulation factor IXa gains proteolytic efficiency upon binding to a phospholipid membrane. We have found that an amphipathic, membrane-binding peptide from the C2 domain of factor VIII, fVIII(2303)(-23), enhances proteolytic efficiency of factor IXa in the absence of phospholipid membranes. This enhancement is the result of a reduction in the K(M) for the substrate, factor X, with little effect on the k(cat). Enhanced function requires interaction of the gamma-carboxyglutamic acid (Gla) domains of factor IXa and factor X since (i) a synthetic peptide comprising the Gla domain of factor IXa and antibodies directed to the Gla domain of factor IXa inhibit this acceleration, (ii) the acceleration is Ca(II) dependent, and (iii) conversion of Gla-domainless factor X is not affected by the presence of fVIII(2303)(-23). The effect of fVIII(2303)(-23) on factor IXa parallels the enhanced function produced by phosphatidylserine containing bilayers, and fVIII(2303)(-23) does not further enhance function of factor IXa when phospholipid vesicles are present. The critical feature of fVIII(2303)(-23) is apparently its amphipathic helix-forming structure [Gilbert, G. E., and Baleja, J. D. (1995) Biochemistry 34, 3022-3031] because other alpha helical peptides such as a homologous peptide from the C2 domain of factor V and melittin have similar effects. Diastereomeric analogues of fVIII(2303)(-23) and melittin, which have reduced helical content, do not support factor IXa activity. A truncated peptide of fVIII(2303)(-23) with three C-terminal residues deleted retains alpha-helical content but loses capacity to enhance factor X cleavage, suggesting that a minimum length of alpha-helix is required. Although these results probably do not illuminate the physiologic function of the factor VIII peptide corresponding to fVIII(2303)(-23), they demonstrate a novel, membrane mimetic role of amphipathic helical peptides in supporting function of factor IXa. PMID- 11009615 TI - Novel inactivation of enoyl-CoA hydratase via beta-elimination of 5, 6-dichloro 7,7,7-trifluoro-4-thia-5-heptenoyl-CoA. AB - 5,6-Dichloro-7,7,7-trifluoro-4-thia-5-heptenoyl-CoA (DCTFTH-CoA) is an analogue of a class of cytotoxic 4-thiaacyl-CoA thioesters that can undergo a beta elimination reaction to form highly unstable thiolate fragments, which yield electrophilic thioketene or thionoacyl halide species. Previous work demonstrated that the medium-chain acyl-CoA dehydrogenase both bioactivates and is inhibited by these CoA thioesters through enzyme-catalyzed beta-elimination of the reactive thiolate moiety [Baker-Malcolm, J. F., Haeffner-Gormley, L., Wang, L., Anders, M. W., and Thorpe, C. (1998) Biochemistry 37, 1383-1393]. This paper shows that DCTFTH-CoA can be directly bioactivated by the enoyl-CoA hydratase (ECH) with the release of 1,2-dichloro-3,3,3-trifluoro-1-propenethiolate and acryloyl-CoA. In the absence of competing exogenous trapping agents, DCTFTH-CoA effects rapid and irreversible loss of hydratase activity. The inactivator is particularly effective at pH 9.0, with a stoichiometry approaching 1 mol of DCTFTH-CoA per enzyme subunit. Modification is associated with a new protein-bound chromophore at 360 nm and an increase in mass of 89 +/- 5 per subunit. Surprisingly, ECH exhibiting less than 2% residual hydratase activity retains essentially 100% beta eliminase activity and continues to generate reactive thiolate species from DCTFTH-CoA. This leads to progressive derivatization of the enzyme with additional UV absorbance, covalent cross-linking of subunits, and an eventual complete loss of beta-eliminase activity. A range of exogenous trapping agents, including small thiol nucleophiles, various proteins, and even phospholipid bilayers, exert strong protection against modification of ECH. Peptide mapping, thiol titrations, UV-vis spectrophotometry, and mass spectrometry show that inactivation involves the covalent modification of Cys62 and/or Cys111 of the recombinant rat liver ECH. These data suggest that enoyl-CoA hydratase is an important enzyme in the bioactivation of DCTFTH-CoA, in a pathway which does not require involvement of the medium-chain acyl-CoA dehydrogenase. PMID- 11009616 TI - Basis for half-of-the-site reactivity and the dominance of the K487 oriental subunit over the E487 subunit in heterotetrameric human liver mitochondrial aldehyde dehydrogenase. AB - Human liver mitochondrial aldehyde dehydrogenase is a tetrameric enzyme composed of 4 identical 500 amino acid containing subunits arranged such that the protein is a dimer of dimers. No kinetic evidence for subunit interactions has been reported. However, the enzyme exhibits half-of-the-site reactivity in that there is a pre-steady-state burst of 2 mol of NADH per mole of enzyme. A variant of the enzyme, found in Asian people, contains a lysine rather than a glutamate at position 487. This enzyme has a high K(M) for NAD(+) and a low specific activity. In heterotetramers composed of both subunit types, it appeared that the lysine containing subunit was dominant over the glutamate-containing subunits. To allow for the separation of various heterotetrameric forms of the enzyme, surface residues were changed. Each of the five possible tetrameric forms of the modified enzyme was isolated and characterized with respect to steady-state kinetics and pre-steady-state burst magnitudes. The data best fit a model where in each dimer pair there is one functioning and one nonfunctioning subunit. Further, the lysine subunit affects the properties only of its dimer partner. Residue 487 is located at the dimer interface, and the glutamate forms salt bonds with two arginine residues. One is to Arg(264) in the same subunit; the other is to Arg(475) located in the other subunit. Most likely the presence of a lysine affects these salt bonds so the lysine subunit can cause the other subunit to become essentially nonfunctional. PMID- 11009617 TI - Folding of green fluorescent protein and the cycle3 mutant. AB - Although the correct folding of green fluorescent protein (GFP) is required for formation of the chromophore, it is known that wild-type GFP cannot mature efficiently in vivo in Escherichia coli at 37 degrees C or higher temperatures that the jellyfish in the Pacific Northwest have never experienced. Recently, by random mutagenesis by the polymerase chain reaction (PCR) method, a mutant called Cycle3 was constructed. This mutant had three mutations, F99S, M153T, and V163A, on or near the surface of the GFP molecule and was able to mature correctly even at 37 degrees C [Crameri et al. (1996) Nat. Biotechnol. 143, 315-319]. In the present study, we investigated the differences in their folding behavior in vitro. We observed the folding and unfolding reactions of both wild-type GFP and the Cycle3 mutant by using green fluorescence as an indicator of the formation of the native structure and examining hydrogen-exchange reactions by Fourier transform infrared spectroscopy. Both proteins showed unusually slow refolding and unfolding rates, and their refolding rates were almost identical under the native state at 25 and at 35 degrees C. On the other hand, aggregation studies in vitro showed that wild-type GFP had a strong tendency to aggregate, while the Cycle3 mutant did not. These results indicated that the ability to mature efficiently in vivo at 37 degrees C was not due to the improved folding and that reduced hydrophobicity on the surface of the Cycle3 mutant was a more critical factor for efficient maturation in vivo. PMID- 11009618 TI - Contribution of individual disulfide bonds to the oxidative folding of ribonuclease A. AB - The eight cysteine residues of ribonuclease A form four disulfide bonds in the native protein. We have analyzed the folding of three double RNase A mutants (C65A/C72A, C58A/C110A, and C26A/C84A, lacking the C65-C72, C58-C110, and C26-C84 disulfide bonds, respectively) and two single mutants (C110A and C26A), in which a single cysteine is replaced with an alanine and the paired cysteine is present in the reduced form. The folding of these mutants was carried out in the presence of oxidized and reduced glutathione, which constitute the main redox agents present within the ER. The use of mass spectrometry in the analysis of the folding processes allowed us (i) to follow the formation of intermediates and thus the pathway of folding of the RNase A mutants, (ii) to quantitate the intermediates that formed, and (iii) to compare the rates of formation of intermediates. By comparison of the folding kinetics of the mutants with that of wild-type RNase A, the contribution of each disulfide bond to the folding process has been evaluated. In particular, we have found that the folding of the C65A/C72A mutant occurs on the same time scale as that of the wild-type protein, thus suggesting that the removal of the C65-C72 disulfide bond has no effect on the kinetics of RNase A folding. Conversely, the C58A/C110A and C26A/C84A mutants fold much more slowly than the wild-type protein. The removal of the C58-C110 and C26-C84 disulfide bonds has a dramatic effect on the kinetics of RNase A folding. Results described in this paper provide specific information about conformational folding events in the regions involving the mutated cysteine residues, thus contributing to a better understanding of the complex mechanism of oxidative folding. PMID- 11009619 TI - Phosphoisoprenoid binding specificity of geranylgeranyltransferase type II. AB - Geranylgeranyltransferase type II (GGTase-II) modifies small monomeric GTPases of the Rab family by attaching geranylgeranyl moieties onto two cysteines of their C terminus. We investigated to what extent GGTase-II discriminates between its native substrate geranylgeranyl pyrophosphate (GGpp) and other phosphoisoprenoids, including farnesyl pyrophosphate (Fpp). On the basis of a novel fluorescent assay, we demonstrated that GGpp binds to GGTase-II with an affinity of 8 +/- 4 nM, while Fpp is bound less strongly (K(d) = 60 +/- 8 nM). Analysis of the binding kinetics of four different phosphoisoprenoids indicated that in all cases association is rapid, with rate constants in the range of 0.15 nM(-1) s(-1). In contrast, the dissociation rates differed greatly, depending on the phosphoisoprenoid used, with weak binding substrates generally displaying an increased rate of dissociation. The affinity of GGpp and Fpp for GGTase-II was also determined in the presence of the Rab7-REP-1 complex. The affinity for GGpp was essentially unaffected by the presence of the complex; Fpp on the other hand bound less strongly to the GGTase-II under these conditions, resulting in a K(d) of 260 +/- 60 nM. In vitro prenylation experiments were used to establish that Fpp not only does bind to GGTase-II but also is transferred with an observed rate constant of 0.082 s(-1) which is very similar to that of GGpp. The implications of the low level of discrimination by GGTase-II for the in vivo specificity of the enzyme and the use of farnesyltransferase inhibitors in anti-cancer therapy are discussed. PMID- 11009620 TI - Vinca alkaloid-induced tubulin spiral formation correlates with cytotoxicity in the leukemic L1210 cell line. AB - The ability of a class of C-20' modified vinca alkaloid congeners to induce tubulin spiral formation was investigated relative to their ability to inhibit microtubule assembly, their cytotoxicity against a leukemic cell line, L1210, and their measured and calculated partition coefficients. These studies were prompted by the observation that the energetics of vinca alkaloid-induced tubulin spiral polymers, or spiraling potential, is inversely related to their clinical dosage and are aimed at the long-term goal of developing the ability to predict the cytotoxic and antineoplastic properties of antimitotic drugs. We demonstrate here that vinca-induced tubulin-spiraling potential is significantly correlated with cytotoxicity against L1210 cells. This is consistent with the size of spirals formed being proportional to the relaxation time for polymer redistribution, the lifetime of cell retention, and effects on microtubule ends and dynamics. Spiraling potential also correlates with calculated but not measured partition coefficients. Surprisingly, spiraling potential does not correlate with the ability to inhibit microtubule formation with purified tubulin or microtubule protein. For the set of C-20' modified compounds studied, the largest inhibitory effects on spiraling potential and cytotoxicity are caused by multiple sites of halogen (-F, -Cl) substitution with the introduction of increased rigidity in the ring. This suggests the C-20' position interacts with a hydrogen bond acceptor or an electrophilic region on the protein that electrostatically disfavors halogen substitutions. These studies are discussed in terms of the cellular mode of action of antimitotic drugs, particularly the importance of microtubule dynamics during mitosis and the factors that regulate those dynamics. PMID- 11009621 TI - Pressure effect on the conformational fluctuation of apomyoglobin in the native state. AB - We have investigated the effect of pressure on fluctuations of the native state of sperm whale apomyoglobin (apoMb) by H/D exchange, fluorescence, and limited proteolysis. The results from intrinsic fluorescence showed that a large fraction of apoMb molecules is in the native conformation in the pressure range from 0.1 to 150 MPa at 293 K and pH 6.0. The H/D exchange of protons of the individual backbone amino acids in this pressure range was monitored by NMR. The rate of H/D exchange was enhanced at high pressure, with the protection factors for some residues decreasing by factors of more than 100 compared to the values at 0.1 MPa. The amplitude of the decrease of the protection factor varied among the individual amino acids on the same secondary structure unit. This result suggests that H/D exchange in apoMb is explained best by the penetration model, in which solvent penetrates into the protein matrix via small motions. The result from limited proteolysis under high pressure showed that a pressure increase does not induce local unfolding of the secondary structure units of apoMb. Conformational fluctuations much smaller than local unfolding evidently provide pathways for water to diffuse into the protein interior, and are enhanced by an increase of pressure. PMID- 11009622 TI - Isotope effects in the transient phases of the reaction catalyzed by ethanolamine ammonia-lyase: determination of the number of exchangeable hydrogens in the enzyme-cofactor complex. AB - Transient phases of the reaction catalyzed by ethanolamine ammonia-lyase (EAL) from Salmonella typhimurium have been investigated by stopped-flow visible spectrophotometry and deuterium kinetic isotope effects. The cleavage of adenosylcobalamin (coenzyme B(12)) to form cob(II)alamin (B(12r)) with ethanolamine as the substrate occurred within the dead time of the instrument whenever coenzyme B(12) was preincubated with enzyme prior to mixing with substrate. The rate was, however, slowed sufficiently to be measured with perdeutero ethanolamine as the substrate. Optical spectra indicate that, during the steady states of the reactions with ethanolamine and with S-2-aminopropanol as substrates, approximately 90% of the active sites contain B(12r). Reformation of the carbon-cobalt bond of the cofactor occurs following depletion of substrate in the reaction mixtures, and the rate constant for this process reflects k(cat) of the respective substrates. This late phase of the reaction also exhibits (2)H isotope effects similar to those measured for the overall reaction with (2)H labeled substrates. With unlabeled substrates, the rate of cofactor reassembly is independent of the number of substrate molecules turned over in the steady-state phase. However, with (2)H-labeled substrates, kinetic isotope effects appear in the reassembly phase, and these isotope effects are maximal after only approximately 2 equiv of substrate/active site are processed. With 5'-deuterated coenzyme B(12) and deuterated substrate, the isotope effect on reassembly is independent of the number of substrate molecules that are turned over. These results indicate that the pool of exchangeable hydrogens in the enzyme-cofactor complex is two-a finding consistent with the hydrogens in the C5' methylene of coenzyme B(12). PMID- 11009623 TI - Structural effects of O-glycosylation on a 15-residue peptide from the mucin (MUC1) core protein. AB - To study the effect of O-glycosylation on the conformational propensities of a peptide backbone, the 15-residue peptide PPAHGVTSAPDTRPA (PPA15) from the MUC1 protein core and its analogue PPA15(T7), glycosylated with alpha-N acetylgalactosamine on Thr7, were prepared and investigated by NMR spectroscopy. The peptide contains both the GVTSAP sequence, which is an effective substrate for GalNAc-T1 and -T3 transferases, and the PDTRP fragment, which is a well-known immunodominant epitope recognized by several anti-MUC1 monoclonal antibodies. Useful structural results were obtained in water upon decreasing the temperature to 5-10 degrees C. The sugar attachment slightly affected the conformational equilibrium of the peptide backbone near the glycosylated Thr7 residue. The clustering of low-energy conformations for both PPA15 and PPA15(T7) within the GVTSAP and APDTRP fragments revealed structural similarities between glycosylated and nonglycosylated peptides. For the GVTSAP region, minor but distinct clusters formed by either PPA15 or PPA15(T7) conformers showed distinct structural propensities of the peptide backbone specific for either the nonglycosylated or the glycosylated peptide. The peptide backbone of the APDTRP fragment, which is a well-known immunodominant region, resembled an S-shaped bend. A similar structural motif was found in the GVTSAP fragment. The S-shaped structure of the peptide backbone is formed by consecutive inverse gamma-turn conformations partially stabilized by hydrogen bonding. A comparison of the solution structure of the APDTRP fragment with a crystal structure of the MUC1 peptide antigen bound to the breast tumor-specific antibody SM3 demonstrated significant structural similarities in the general shape. PMID- 11009624 TI - Calcium enhances heparin catalysis of the antithrombin-factor Xa reaction by promoting the assembly of an intermediate heparin-antithrombin-factor Xa bridging complex. Demonstration by rapid kinetics studies. AB - Heparin catalyzes the inhibition of factor Xa by antithrombin mainly through an allosteric activation of the serpin inhibitor, but an alternative heparin bridging mechanism has been suggested to enhance the catalysis in the presence of physiologic calcium levels due to calcium interactions with the Gla domain exposing a heparin binding exosite in factor Xa. To provide direct evidence for this bridging mechanism, we studied the heparin-catalyzed reaction of antithrombin with factor Xa, Gla-domainless factor Xa (GDFXa), and a heparin binding exosite mutant of GDFXa in the absence and presence of calcium using rapid kinetic methods. The pseudo-first-order rate constant for factor Xa inhibition by antithrombin complexed with a long-chain approximately 70 saccharide heparin showed a saturable dependence on inhibitor concentration in the presence but not in the absence of 2.5 mM Ca(2+), indicating the formation of an intermediate heparin-serpin-proteinase encounter complex with a dissociation constant of approximately 90 nM prior to formation of the stable serpin proteinase complex with a rate constant of approximately 20 s(-1). Similar saturation kinetics were observed for the inhibition of GDFXa by the antithrombin heparin complex, except that Ca(2+) was not required for the effect. By contrast, no Ca(2+)-dependent saturation of the inhibition rate constant was detectable over the same range of inhibitor concentrations for reactions of either a short chain approximately 26-saccharide high-affinity heparin-antithrombin complex with factor Xa or the long-chain heparin-antithrombin complex with the heparin binding exosite mutant, GDFXa R240A. These findings suggest that binding of full-length heparin chains to an exosite of factor Xa in the presence of Ca(2+) produces a chain-length-dependent lowering of the dissociation constant for assembly of the intermediate heparin-antithrombin-factor Xa encounter complex, resulting in a several 100-fold rate enhancement by a heparin bridging mechanism. PMID- 11009625 TI - Effects of compound structure on carbazole dication-DNA complexes: tests of the minor-groove complex models. AB - Carbazole dications have shown excellent activity against opportunistic infections, but they are quite different in structure from previously studied unfused aromatic cations that function by targeting the DNA minor groove. In a previous report [Tanious, F. A., Ding, D., Patrick, D. A., Tidwell, R. R., and Wilson, W. D. (1997) Biochemistry 36, 15315-15325] we showed that, despite their fused ring structure, the carbazoles also bind in A/T sequences of the DNA minor groove and we proposed models for the carbazole-DNA complexes with the carbazole nitrogen facing out of the groove for 3,6 substituted compounds but into the groove in 2,7 carbazoles. To test and refine the models, carbazole-N-methyl substituted derivatives have been synthesized in both the 3,6 and 2,7 series as well as a new 2,6 substituted NH derivative that is intermediate in structure. Footprinting results indicate a broad AT specificity of carbazole binding and a pattern in agreement with a minor groove complex. Surface plasmon resonance biosensor analysis of carbazole binding to an oligomer with an AATT central sequence indicated that the 2,7 NH compound has the largest binding constant. Both the 3,6 NH and NMe compounds bind with similar equilibrium constants that are less than for the 2,7 NH compound. The 2,7 NMe compound has the lowest binding constant of all the carbazoles. Spectroscopic results are also similar for the two 3,6 derivatives but are quite different for the 2,7 NH and NMe carbazole dications. Structural analysis of carbazole complexes with an AATT sequence by 2D NMR methods also supported a minor groove complex of the carbazoles in orientations in agreement with the previously proposed models. From these results, it is clear that the fused ring carbazoles can bind strongly in the DNA minor groove with a broad A/T specificity and that the 2,7 and 3,6 substituted carbazoles bind to the minor groove in opposite orientations. PMID- 11009626 TI - Intramolecular activation of a Ca(2+)-dependent protein kinase is disrupted by insertions in the tether that connects the calmodulin-like domain to the kinase PMID- 11009627 TI - A computer model of normal conduction in the human atria. AB - Although considerable progress has been made in understanding the process of wavefront propagation and arrhythmogenesis in human atria, technical concerns and issues of patient safety have limited experimental investigations. The present work describes a finite volume-based computer model of human atrial activation and current flow to complement these studies. Unlike previous representations, the model is three-dimensional, incorporating both the left and right atria and the major muscle bundles of the atria, including the crista terminalis, pectinate muscles, limbus of the fossa ovalis, and Bachmann's bundle. The bundles are represented as anisotropic structures with fiber directions aligned with the bundle axes. Conductivities are assigned to the model to give realistic local conduction velocities within the bundles and bulk tissue. Results from simulations demonstrate the role of the bundles in a normal sinus rhythm and also reveal the patterns of activation in the septum, where experimental mapping has been extremely challenging. To validate the model, the simulated normal activation sequence and conduction velocities at various locations are compared with experimental observations and data. The model is also used to investigate paced activation, and a mechanism of the relative lengthening of left versus right stimulation is presented. Owing to both the realistic geometry and the bundle structures, the model can be used for further analysis of the normal activation sequence and to examine abnormal conduction, including flutter. The full text of this article is available at http://www.circresaha.org. PMID- 11009628 TI - Long-distance learning. PMID- 11009629 TI - Patient rights reach Capitol Hill. PMID- 11009630 TI - Compliance efforts get some PEPP. PMID- 11009631 TI - CPR systems: which one is right for your organization? PMID- 11009632 TI - Next generation: how Internet technology propels the electronic medical record. PMID- 11009633 TI - Beyond Y2K: no rest for the weary. PMID- 11009634 TI - Tools for defining data. PMID- 11009635 TI - PPS + new technology = growing pains. PMID- 11009636 TI - Reimbursement at your fingertips--a glossary of terms. PMID- 11009637 TI - Practice brief. Correcting and amending entries in a computerized patient record. American Health Information Management Association. PMID- 11009638 TI - Breaking with tradition: improving HIM functions in behavioral health. PMID- 11009639 TI - Using electronic resources to recruit and hire employees. PMID- 11009640 TI - Getting ready for APCs. PMID- 11009641 TI - Summary of ICD-9-CM Coordination and Maintenance Committee meeting. AB - The ICD-9-CM Coordination and Maintenance Committee, cosponsored by the National Center for Health Statistics (NCHS) and the Health Care Financing Administration (HCFA), recently met in Baltimore, MD. Donna Pickett, RRA (NCHS), and Patricia Brooks, RRA (HCFA), cochaired the meeting. Proposed modifications to ICD-9-CM were presented and are summarized below. Unless otherwise indicated, the audience generally supported the proposed changes. PMID- 11009642 TI - Charting new territory. PMID- 11009643 TI - HSS proposes HIPAA privacy standards. PMID- 11009644 TI - Using CPT modifier -25 for professional billing. PMID- 11009645 TI - E-commerce and HIM: ready or not, here it comes. PMID- 11009646 TI - A crystal ball for coding. PMID- 11009647 TI - XML: defining the transition from paper to digital records. PMID- 11009648 TI - Trend watch: coding for nonphysician services. PMID- 11009649 TI - Coders join forces with physicians to improve clinical outcomes. PMID- 11009650 TI - An off-site coding success story. PMID- 11009651 TI - Ten down under: implementing ICD-10 in Australia. PMID- 11009652 TI - A global language for pharmaceutical regulation. PMID- 11009653 TI - Practice brief. Information security: a checklist for healthcare professionals (updated). American Health Information Management Association. PMID- 11009654 TI - When you don't see eye to eye with a Joint Commission surveyor. PMID- 11009655 TI - Using scenario planning. PMID- 11009656 TI - CPT changes for 2000. PMID- 11009657 TI - Incentive programs offer aid to increase coding productivity. PMID- 11009658 TI - American Health Information Management Association. Standards of ethical coding. PMID- 11009659 TI - Ensuring compliance in the new frontier. PMID- 11009660 TI - AHIMA comments on privacy rule. PMID- 11009661 TI - Medical and clinical service contracts: a compliance focus. PMID- 11009662 TI - Web-based patient record systems: opportunities for best practices. PMID- 11009663 TI - For health records on the Internet, the future is now. PMID- 11009664 TI - Online consumer health records: revolution or confusion? PMID- 11009665 TI - PCASSO: a model for safe use of the Internet in healthcare. PMID- 11009666 TI - Intranets: a way to connect. PMID- 11009667 TI - "Pointing and clicking" your way to knowledge. PMID- 11009668 TI - Web-based education cuts training time, saves money. PMID- 11009669 TI - Patient-centered e-mail: developing the right policies. PMID- 11009670 TI - Simplified deficiency processing brings hospital-wide benefits. PMID- 11009671 TI - Resolving confidentiality barriers in research data collection. PMID- 11009672 TI - Professional practice solutions ... hospital scheduled to be surveyed this year by the Joint Commission. PMID- 11009673 TI - Post-acute markets face Medicare, HCFA changes. PMID- 11009674 TI - Practice brief. Authentication of health record entries (updated). American Healthcare Information Management Association. PMID- 11009675 TI - Hiring the best: testing applicants. PMID- 11009676 TI - Committee proposes ICD-9-CM changes for 2001. PMID- 11009677 TI - Reviewing the details of coding septicemia. PMID- 11009678 TI - IOM report examines medical errors. PMID- 11009679 TI - HIPAA: understanding the requirements. PMID- 11009680 TI - Taking the lead in compliance education. PMID- 11009681 TI - Are we ready for portable healthcare? PMID- 11009682 TI - HIPAA: where to begin? Next steps? First steps? Getting a grip on HIPAA security standards. Panel discussion. PMID- 11009683 TI - Getting ready for HIPAA privacy rules. PMID- 11009684 TI - Health informatics standards: a user's guide. PMID- 11009685 TI - Long-term care's prospects for accreditation. PMID- 11009686 TI - To "e" or not to "e": HIM and the dawn of e-health. PMID- 11009687 TI - MPIs (master patient index) in healthcare: current trends and practices. PMID- 11009688 TI - Boosting efficiency in home health record systems. PMID- 11009689 TI - Practice brief. Destruction of patient health information (updated). American Health Information Management Association. PMID- 11009690 TI - Guidelines for a "win-win" vendor relationship. PMID- 11009691 TI - Clarifying selected CPT modifiers. AB - This discussion includes only 20 percent of the modifiers available for physician reporting, but could account for 80 percent of modifiers assigned for physician services. Modifier -91 was the only new modifier introduced in CPT 2000 and it is used to report a repeat clinical diagnostic laboratory test. It is not to be used when confirming initial results or for any other reason other than a clinical need to repeat the test for the same patient on the same day. Success with CPT modifier reporting requires a thorough review of CPT guidelines and some detective work for identifying health plan requirements for modified codes. PMID- 11009692 TI - Modifier usage for hospital outpatient services. PMID- 11009693 TI - Setting the standards. PMID- 11009694 TI - Blueprint for a 21st century hospital: a casebook in strategic and operational planning. PMID- 11009695 TI - Why patients complain and what health professionals can do about it. PMID- 11009696 TI - Reducing medication errors. Part II: Newton-Wellesley Hospital, Newton, MA. PMID- 11009697 TI - Gearing up for the JCAHO's new pain management standards: some hospitals are ahead of the game. PMID- 11009698 TI - Grassroots decisionmaking informs care redesign at York Hospital in York, PA. PMID- 11009699 TI - Katie's crusade. Colon cancer is one of the deadliest and most preventable malignancies. What you need to know about the disease--and the surprisingly painless test that could save your life. PMID- 11009700 TI - Spare a kidney? You don't have to be dead to donate an organ, but you had better know the risks before you give it up. PMID- 11009702 TI - A pencil in his heart. Nathan king is alive today because his mother was level headed enough not to try to pull it out. PMID- 11009703 TI - Smoke screen. Kids are less likely to use tobacco if they're shown how cigarette companies try to dupe them. PMID- 11009701 TI - When pills make sense. Some parents turn too quickly to mood-altering drugs. But often medication is the right choice. PMID- 11009704 TI - The dizzy mystery. Dizzy spells are a natural consequence of aging, but they can also mean that something is wrong. PMID- 11009705 TI - Affair of the heart. A new study finds that estrogen may not protect women against cardiac disease. Here's why. PMID- 11009706 TI - Cloning the new babes. Science comes closer to creating the perfect pig, offering hope for a larder of rejection-free organs. PMID- 11009708 TI - A biotech wreck. It's an industry full of Ps--promises and potential. Call me when they get to profits. PMID- 11009707 TI - The Web docs. The Internet can fill the gaps in your medical knowledge. But you need to know how to separate the good sites from the bad. PMID- 11009709 TI - Diabetes recall. Though Rezulin's withdrawal is a blow for some patients, there are ways to cope. Here's how. PMID- 11009710 TI - Cutting stealth flab. That winter baggage may be less than you thought. Still, it's spring, so here's how to lose it. PMID- 11009711 TI - The Feds step up the pace. In the hot race to wrap up the human genome, government mappers say they're two-thirds home. PMID- 11009712 TI - Unkindest cut? Most moms don't need a minor surgery to deliver. PMID- 11009713 TI - Will the mind figure out how the brain works? PMID- 11009714 TI - Will we figure out how life began? PMID- 11009716 TI - Will we keep evolving? PMID- 11009715 TI - Will we clone a dinosaur? PMID- 11009717 TI - The race is over. The great genome quest is officially a tie, thanks to a round of pizza diplomacy. Yet lead researcher Craig Venter still draws few cheers from his colleagues. PMID- 11009718 TI - The genome is mapped. Now what? It will be decades before scientists identify and understand all of our genes. But that hasn't stopped them from making dramatic discoveries. PMID- 11009719 TI - The double helix revisited. The man who launched the Human Genome Project celebrates its success. PMID- 11009720 TI - No doctor required. The FDA may expand the number of prescription drugs it converts to over-the-counter use. PMID- 11009721 TI - The new science of Alzheimer's. Racing against time--and one another--researchers close in on the aging brain's most heartbreaking disorder. PMID- 11009722 TI - Hot-flash relief. The list of alternatives to hormone-replacement therapy keeps growing. What's a woman to do? PMID- 11009723 TI - An Rx for pills--and politics. What's hot this election year? Drug costs and a pair of thorny tax issues. PMID- 11009724 TI - Little hope, less help. The epidemic has hit with devastating force--and things will get much worse before they get better. PMID- 11009725 TI - The politics of AIDS. When the President is a dissident. PMID- 11009726 TI - The new antiseptics. Two novel approaches to an ancient scourge could someday protect millions from runaway infections. PMID- 11009727 TI - Grains of hope. Genetically engineered crops could revolutionize farming. Protesters fear they could also destroy the ecosystem. You decide. PMID- 11009728 TI - Ginseng surprise. An independent report finds that what's listed on a supplement's label is not always what's inside. PMID- 11009729 TI - Coordination of care: stage one in assessing rural managed care. AB - This article provides a framework for assessing the role of managed care in optimizing the mix of services to advance the health of rural populations. Mindful of limited organizational and financial resources in many rural areas, such assessments must carefully weigh the contribution of these resources to effective management of care. The framework presented emphasizes the different populations and different health conditions that a variety of care coordinators may address more or less effectively. Attention to issues having to do with organizational architecture and financial accountability of managed care should flow from prior to consideration of care management. PMID- 11009730 TI - Comparative performance assessment in managed care: data envelopment analysis for health care managers. AB - In health care markets increasingly dominated by managed care, health care managers need sophisticated yet understandable comparative performance assessment methods. Ideally, these methods should provide targets for the managers to improve program or organization performance. This article reviews three comparative performance methodologies with a comprehensive explanation of data envelopment analysis (DEA). DEA is a form of linear programming that allows for the inclusion of multiple input and output/outcome performance variables from a virtually unlimited number of organizations, facilitating meaningful health service provider comparative assessments. PMID- 11009731 TI - Managed care liability and ERISA preemption. AB - Patients enrolled in managed care organizations (MCOs) are concerned about quality of care and restrictions imposed. There is concern about being harmed due to the negligence of MCOs. Meanwhile, MCOs are protected by the federal Employment Retirement Income Security Act (ERISA) of 1974, which preempts state law claims and results in eliminating many of the recoveries otherwise available to a harmed beneficiary in non-MCOs. This article reviews the ERISA exemption and legal theories for patient redress. PMID- 11009732 TI - "Low-tech" personal emergency response systems reduce costs and improve outcomes. AB - A relatively inexpensive low-technology solution can be used by managed care organizations (MCOs) to improve outcomes and significantly reduce health care costs among community-residing elderly patients. Clinical studies indicate that usage of monitored Personal Emergency Response Systems (PERS) reduce mortality rates by nearly four times, reduce hospital utilization by 59 percent, and yield a positive benefit-to-cost ratio of over seven to one (every dollar spent on PERS results in $7.19 in health care cost savings). PMID- 11009733 TI - Don't ask, don't tell: the incontinence conspiracy. AB - Don't ask, don't tell is the attitude of both patients and doctors toward symptoms of urinary incontinence (UI). This has serious fiscal implications for managed care, because the consequences of not treating the condition increase the annual cost of care by an estimated $3,941 per individual. Behavioral treatments have clinical efficacy of 87 percent, and should be tried first, according to the clinical practice guidelines published by the Agency for Health Care Policy and Research. Attitudes about the condition, and historical and current interventions, are discussed. PMID- 11009734 TI - Information system powers innovative research project aimed at managing the care of senior citizens. AB - HealthAmerica Pennsylvania, Inc., in Pittsburgh, is aiming to increase access to health care for senior citizens in its Advantra health plan in Pittsburgh. Under its Senior Life Management program, Coventry Health Care, parent company of HealthAmerica, hired nine personal service representatives (PSRs) and is asking them to provide outreach to 5,000 senior citizens. The PSRs and other staff members will get to know each senior citizen in the program, evaluate and recognize their health needs, and develop methods to ensure that their needs are met before any patient's condition becomes unnecessarily costly. The goal of this innovative program is to improve access to health care for these 5,000 members and thus improve their satisfaction with care and help to reduce health care costs. PMID- 11009735 TI - Interview with Bruce Vladeck, Institute for Medicare Practice. PMID- 11009736 TI - Determinants of specialty physician practice management integration. AB - Physicians increasingly organize strategically to mitigate the potentially adverse impacts of managed care on practice incomes. There are alternative routes to physician organizing. This article used a 1996-1997 survey of orthopedics and obstetrics-gynecology (OB-GYN) practices in Memphis, Tennessee, to investigate the determinants of specialty physician integration through physician practice management (PPM) firms. The percentage of managed care contracts indicates a weak but positive correlation with PPM integration. Different factors appear to influence the likelihood of integration in different specialties. PMID- 11009737 TI - The radical revolution of radiology. AB - The radiologic segment of the health care industry has grown dramatically in size and functions. The focus on cancer and heart disease has made it a key core functional area for health systems and physician groups. Aggressive growth plans are based on key strategies, high demand services, and other critical success factors. PMID- 11009739 TI - Understanding autism. PMID- 11009738 TI - Right off the shelf. Non-prescription cholesterol drugs could open an era of do it-yourself medicine. PMID- 11009740 TI - Proposed PPS refinements would boost payments for sickest patients; add no new Medicare money to system. PMID- 11009741 TI - How to use the MDS quality indicators to improve patient care. PMID- 11009742 TI - Industry reaction to the PPS refinement proposal is mixed. PMID- 11009743 TI - The future of health status assessment. PMID- 11009744 TI - Regulating the pharmaceutical industry: who really benefits? PMID- 11009745 TI - Can peer-comparison feedback improve patient functional status? AB - OBJECTIVE: To determine whether providing physicians with peer-comparison feedback can improve patient functional status. STUDY DESIGN: Randomized, controlled, comparative study. METHODS: Forty-eight primary care physicians at Kaiser Permanente Woodland Hills, a group-model health maintenance organization in southern California, were randomly assigned to an intervention group or a control group. All physicians were informed that their elderly patients (randomly selected patients aged 65 to 75) would be monitored. Physicians in the intervention group received aggregated peer-comparison feedback data (physician "report cards") on the functional status of their elderly patients. Physicians in the control group received only general information that their patients' functional status would be monitored. The effect of the intervention on patients' functional status was determined by comparing responses to surveys completed by the patients at baseline and after the intervention. RESULTS: Patients in both the control and intervention groups had a statistically significant decrease in functional status, including decreases in their ability to complete daily activities and increases in pain. In addition, patients in the control group reported a significant decrease in social activities, physical fitness, and feelings. In the intervention group, patients also experienced a significant decrease in social support. CONCLUSIONS: Educational interventions, including peer-comparison feedback, did not result in improvements in patient functional status. Research is desperately needed to identify interventions that can lead to improved health for elderly patients. PMID- 11009746 TI - Preventive service use and Medicaid managed care in New York City. AB - OBJECTIVE: To examine the effect of managed care enrollment on the use of preventive services among New York City's Medicaid population. STUDY DESIGN: An analysis of survey results from a sample of Medicaid beneficiaries in managed care plans and in traditional Medicaid. METHODS: This study is based on a 1994 survey of 1038 Medicaid beneficiaries enrolled in any of 5 managed care plans and a comparison group of 410 beneficiaries in traditional Medicaid in New York City. The survey data are used to examine the effect of managed care on the self reported use of Pap smears, mammograms, and infant immunizations. We performed bivariate analysis to compare the use of preventive services between managed care enrollees and beneficiaries in traditional Medicaid. We also used multivariate logistic analysis to explore this comparison, controlling for factors that may confound the relationship. RESULTS: Medicaid beneficiaries in managed care were no more or less likely to receive infant immunizations, Pap smears, or mammograms than those in the traditional Medicaid program. CONCLUSIONS: Our analysis suggests that Medicaid managed care and the traditional program performed the same in getting appropriate preventive services to beneficiaries. PMID- 11009747 TI - The impact of health insurance market reforms on market competition. AB - OBJECTIVES: To assess the impact of state and federal health insurance market reforms on the nature and extent of market competition. STUDY DESIGN: Qualitative, comparative case studies in 7 states. METHODS: Two rounds of in depth interviews were conducted with over 100 key informants from the insurance industry. In each state, these sources included 2 to 4 regulators, 5 to 6 independent agents, and several sources at each of 4 to 5 of the top insurers. Extensive documentary data relating to market activity were also collected. These multiple sources of information and data were analyzed with both qualitative and quantitative techniques. RESULTS: (1) Small-group health insurance markets are highly competitive, both in price and in product innovation and diversity. (2) In some of the more heavily regulated states, there is very little competition in less-populated areas, especially for indemnity insurance. (3) The rapid growth of managed care in the small-group market may have been precipitated by these reforms. (4) Standardized benefit plans have not achieved their objectives. (5) Competitive forces still focus to a considerable extent on risk selection techniques. CONCLUSION: Small-group market reforms have not harmed market competition and may have improved competition in several respects. However, these reforms do not alter the fundamental orientation of competitive insurance markets, which is to focus on risk selection factors and techniques to the extent feasible. PMID- 11009748 TI - An instrument to measure patient satisfaction with healthcare in an observational database: results of a validation study using data from CaPSURE. AB - OBJECTIVE: To validate a satisfaction measure for use in longitudinal, prospective studies of patient care. STUDY DESIGN: Patients with biopsy-confirmed prostate cancer (n = 228) who were enrolled in CaPSURE (Cancer of the Prostate Strategic Urologic Research Endeavor) completed a self-administered questionnaire that included a health-related quality-of-life and satisfaction measure. A subset of patients completed the questionnaire again within 30 days. METHODS: The satisfaction measure contained 6 individual subscales: overall satisfaction with care, contact with providers, confidence in providers, communication skills, humaneness, and a summary scale. Six items surveyed patients' willingness to participate in decision making (participatory style), and these were averaged into a single score. Variability, reliability, stability, and validity were evaluated. RESULTS: Responses to the items varied substantially. The overall satisfaction scale demonstrated good internal consistency reliability (Cronbach alpha = 0.82) and moderate test-retest reliability (0.62), and it could discriminate between groups of individuals expected to differ with regard to satisfaction (by age and disease stage). Subscale internal consistency reliability (0.37-0.54) and stability (0.38-0.63) were weaker, suggesting that only a single scale should be reported. The participatory scale performed poorly and could not be recommended for future use. CONCLUSION: The overall satisfaction measure developed for this study demonstrated good reliability and validity and should be useful in other population-based studies in conjunction with other outcome measures. PMID- 11009749 TI - Acne vulgaris: pathogenesis and therapeutic approach. AB - AUDIENCE: This activity is designed for primary care providers, internists, and general audiences. GOAL: To provide the reader with a basic understanding of the pathogenesis of acne vulgaris and the rationale behind several treatment options. Clinicians should be aware of treatment strategies for various subtypes of acne vulgaris. OBJECTIVES: 1. To describe the emotional and financial impact of acne. 2. To describe the differential diagnosis of acne vulgaris. 3. To discuss acne's pathogenesis. 4. To provide information on topical therapy for the treatment of acne vulgaris. 5. To discuss oral therapy for the treatment of acne vulgaris. PMID- 11009750 TI - Parkinson's disease: focus on management alternatives. AB - AUDIENCE: This activity is designed for physicians, pharmacists, nurses, health planners, directors of managed care organizations, and payers of health services. GOAL: To understand the impact that Parkinson's disease has on patients and to identify areas of drug therapy that can be optimized to improve a patient's quality of life. OBJECTIVES: 1. Discuss the epidemiology and pathophysiology of idiopathic Parkinson's disease (IPD). 2. Identify causes of secondary parkinsonism and medications associated with drug-induced parkinsonism. 3. Discuss the cardinal and secondary manifestations of IPD. 4. Outline the 6 stages of IPD. 5. Identify therapeutic alternatives for various levodopa treatment failures. 6. Identify the 3 newer antiparkinson agents and state their mechanisms of action, common adverse effects, drug interactions, and appropriate use. 7. Identify the appropriate use, common adverse effects, and drug interactions of dopamine agonists, anticholinergic agents, monoamine oxidase inhibitors, and catechol-O-methyltransferase inhibitors in Parkinson's disease therapy. PMID- 11009751 TI - Optimizing quality of care and cost effectiveness in treating allergic rhinitis in a managed care setting. AB - Allergic rhinitis is a common condition in managed care populations. The direct medical cost of rhinitis exceeded $3 billion in 1996, and an additional cost of $4 billion resulted from the exacerbation of other concomitant conditions, such as asthma or otitis media. Costs continued to increase in 1999; sales of prescription antihistamines and nasal steroids exceeded $3 billion and $1 billion, respectively. The indirect costs of allergic rhinitis include lost work productivity, reduced performance and learning, and increased workplace and traffic accidents. Rhinitis treatments include allergen avoidance, over-the counter (OTC) sedating antihistamines, nonsedating antihistamines, nasal steroids, and immunotherapy. Allergen avoidance strategies for patients with asthma and rhinitis are ineffective or are of very limited benefit. Allergists criticize the use of OTC sedating antihistamines, which are associated with reduced learning and performance even when sedation does not occur. Evidence based literature reviews of clinical trials have shown that nasal steroids are more effective than nonsedating antihistamines in the treatment of rhinitis. The most commonly prescribed nasal steroid, fluticasone, has been shown to be effective in treating rhinitis and in improving patients' quality of life. It is also more cost effective than the most commonly prescribed antihistamine, loratadine. Clinical trials have indicated that immunotherapy is expensive and of limited benefit. As these evidence-based findings are used to develop managed care treatment guidelines, nasal steroids are likely to be recommended as the first-line treatment for rhinitis, which should result in lower treatment costs and improved outcomes for patients with rhinitis. PMID- 11009752 TI - Computerization saves money, time, and trees. PMID- 11009753 TI - Center will establish rehab care benchmarks. PMID- 11009754 TI - Why don't doctors follow guidelines? PMID- 11009755 TI - Medication errors top patients' worry list. PMID- 11009756 TI - Tracking your medical errors just became a priority. PMID- 11009757 TI - Two hospitals solve adverse events problems. PMID- 11009758 TI - Don't accept ED's claim it's too busy to benchmark. PMID- 11009759 TI - Teamwork helps cut ED wait times. PMID- 11009760 TI - Benchmarking focuses on perfecting pastoral care. PMID- 11009761 TI - Getting a pastoral care program off the ground. PMID- 11009762 TI - Regional benchmarks show how you stack up. PMID- 11009763 TI - Benchmarking crucial for small, rural facilities with few resources. PMID- 11009764 TI - Monitoring system saves hospital $180,000 a year. PMID- 11009765 TI - Communication eases consolidation conflicts. PMID- 11009766 TI - Climbing hospital stays level off in western U.S. PMID- 11009767 TI - Who will lose under the outpatient PPS? PMID- 11009768 TI - Perspectives. Regression to the mean: PPOs, risk-sharing are market mainstays. PMID- 11009769 TI - Perspectives. Congressional outlook 2000: you snooze, you lose. PMID- 11009770 TI - Perspectives. Surplus makes both 'restraint,' new programs possible in budget proposal. PMID- 11009771 TI - Marketplace. Provider risk-sharing in managed care: still viable despite current unpopularity. PMID- 11009772 TI - Perspectives. The uninsured: local efforts in state, federal, global contexts. PMID- 11009773 TI - Define improvement opportunities to cut 'data denial'. PMID- 11009774 TI - Studies offer benchmarks for costs, rates of prostate surgery. PMID- 11009775 TI - Competing hospitals use regional database to cut cardiac surgery mortality. PMID- 11009776 TI - Canada: using our most valuable natural resource to transform health care. AB - Canada is recognized around the world as a wonderful outdoor experience with its wealth of natural resources and vibrancy. As Canada has evolved through the years, this vibrancy has continued to resonate through its communities and people. Historically, Canadians are well known for their pioneering and frontier spirit, which is a precious commodity. Now, that spirit, wisdom, talent, and potential of Canadians are being tapped to begin to change its health care system. PMID- 11009777 TI - Henk Tjassing: on the road to Suriname. Interview by Margaret J. Cushman. PMID- 11009778 TI - Cultural competence in patient education. AB - As a community health nurse, have you tested your cultural consideration quotient in patient education recently? Can you define the difference between ethnocentrism and multiculturalism? Do you find that you and your colleagues work with more or less transcultural nursing in your daily practice? Find out why cultural competence is vital to nurses' success in the 21st century. PMID- 11009779 TI - A salute to congressman Robert A. Weygand. Advocate for the elderly. Interview by Val J. Halamandaris. PMID- 11009780 TI - The Frances Payne Bolton WHO Collaborating Center: promoting home care globally. AB - Since its designation as a World Health Organization Collaborating Center in 1993, the Frances Payne Bolton School of Nursing has been committed to furthering the development of home care practices throughout the world. Through research, education, conferences, and publications, the Bolton School is promoting the efforts made by home care nurses throughout the world. PMID- 11009782 TI - Hospice forum. Resources for public engagement activities; advances in clinical policy for end-of-life care. PMID- 11009781 TI - Elder suicide: a preventable tragedy. AB - Reported suicides claim more than 32,000 lives annually in the United States. More than 6,200 of these deaths occur among those 65 years of age or older. Suicide is the third leading cause of death among the elderly. These deaths are unnecessary and most are avoidable. Elder suicides happen because not enough is done to prevent them. Home care agencies can do something about this situation. PMID- 11009783 TI - Reaching out ... through communication. PMID- 11009784 TI - PPS: blessing or curse? PMID- 11009785 TI - A conversation with Sandra R. Hernandez, MD. Interview by Val J. Halamandaris. PMID- 11009786 TI - Preventing nursing turnover in home care. AB - Turnover is costly to any organization. Frequently, turnover creates a ripple effect that is felt by the agency, its patients, and staff at all levels. The very nature of home care creates an environment in which nurses are not visible in the office every day. It is difficult to track the activities of new employees on a day-to-day basis to know how they are progressing through the orientation process. What causes turnover among registered nurses within the first six months of employment? One agency shares its story and related findings. PMID- 11009787 TI - Changing times in rehab staffing. AB - Historically, a low supply of physical and occupational therapists existed in the home care marketplace. Home care agencies have had to contract with private practices and accept therapists who could only provide two or three visits a week to keep up with the demand. However, cutbacks have occurred at the acute, sub acute, and rehabilitation levels of inpatient care. Now, many therapists who would not have considered home care employment in the past are asking for interviews. For once, many home care agencies have the opportunity to carefully select the therapists they hire. PMID- 11009788 TI - A tribute to Beverly Malone. Interview by Val J. Halamandaris. PMID- 11009789 TI - The greying of the profession. AB - As the 21st century begins, Americans can expect to see changes across the health care spectrum. With the drive toward market parity in the management of health care resources, it is easy to lose sight of some critical elements of the health care system that may affect the quality and delivery of care. Specifically, the work force that cares for patients in the hospital, the home, or other care settings will shift. The nursing profession will not be immune to this shift and, as nurses reach retirement age and leave the profession, their departure may leave a void that could take years to fill and profoundly alter the quality of care available. PMID- 11009790 TI - Pre-employment intentions of home care aides. AB - The demand for home care aides (HCA) is projected to increase substantially over the next several years. One of the reasons for this projection is the high turnover rates associated with the occupation. Although there are many reasons for the high turnover after HCAs have been hired, their pre-employment intentions are unclear. A better understanding of HCAs' pre-employment intentions may help to decrease recruitment and training costs for home care agencies and improve continuity of care for clients. PMID- 11009791 TI - Determining the scope of practice for home care aides. AB - Because of recent fiscal challenges, home health agencies are forced to seek the most cost-effective methods of providing care. Many agencies are attempting to redefine the scope of practice for home care aides, hoping to determine how to best use these individuals in providing high-quality, cost-effective care. A study was conducted to determine what tasks should be included in the aide's scope of practice. The high response rate to this study seems to indicate a great deal of interest and the need to resolve this issue. PMID- 11009792 TI - Nine tips to improve your recruitment advertising. PMID- 11009793 TI - Humor at work: could this keep staff happy? PMID- 11009795 TI - Wanted ... angels on earth. PMID- 11009794 TI - Fatigue: the most important consideration for the patient with cancer. AB - The Fatigue Coalition--a multidisciplinary group of medical practitioners, researchers, and patient advocates--conducted a study to determine the effects of fatigue on cancer patients. The study, published in September 1998, examined cancer patients, caregivers, and oncologists. This survey presents new information about how fatigue is perceived by the patient, caregiver, and oncologist. In response to the survey, the group developed a series of educational and research initiatives designed to help patients and physicians better understand chemotherapy-related fatigue and provide successful interventions. PMID- 11009796 TI - Integrating effective home care aide recruitment & retention strategies. PMID- 11009797 TI - NAHC's legislative agenda tackles mission-critical issues. PMID- 11009798 TI - The new hospice compliance plan: defining and addressing risk areas. AB - The Department of Health and Human Services' (HHS) Office of the Inspector General (OIG), has advised hospices and other health care providers to formulate effective controls to ensure compliance with federal and state statutes, rules, and regulations, and private-payor health care program requirements. The recently released guidelines provide a blueprint for developing such programs. This is the first of four installments that focus specifically on the 28 risk areas identified in the guidance and offer strategies for incorporating them in a hospice compliance program. The authors have organized the 28 risk areas under 9 topic domains to simplify the task of tackling the guidance. This article covers the first two areas: Admission/Certification/Recertification and Billing. PMID- 11009799 TI - Ensuring accuracy of OASIS data as home care moves toward PPS. PMID- 11009800 TI - Skilled nurses in schools: IDEA (Individuals with Disabilities Education Act) at work. AB - Home care agencies have an opportunity to create a new revenue stream while helping disabled children receive their education within schools, instead of at home, through the Individuals with Disabilities Education Act. This article examines the law itself, related court cases, problems with implementation, and implications for home care providers. Home care agencies must demonstrate to schools that they can deliver the services the schools need to supplement their nursing staffs under IDEA. PMID- 11009801 TI - Reflections on home care. Where we've been, where we're going, why we must get there. PMID- 11009802 TI - Taking a stand. Members of Congress praise home care's benefits importance. PMID- 11009803 TI - The lowdown on colon cancer. PMID- 11009804 TI - Health care delivery at the millennium: 10-year forecast. PMID- 11009805 TI - System losses, poor capitation profitability, bankruptcy warnings signal distress for docs. PMID- 11009807 TI - Don't expect instant savings when implementing Medicaid managed care. PMID- 11009806 TI - Planning for clinical services within a regional health system. PMID- 11009808 TI - New data project median hospital's margin will be -0.41 in 2002 because of BBA's impact. PMID- 11009809 TI - Avoid politics by establishing objective criteria for capital budget decision making. PMID- 11009810 TI - Hospitals face major changes in 2000. PMID- 11009811 TI - Developer offers 'win-win' construction solution for capital-starved hospitals. PMID- 11009812 TI - HIPAA offers hospitals the good, the bad and the ugly. PMID- 11009813 TI - After more major acquisitions, affiliations, Healtheon says big pieces now in place. PMID- 11009814 TI - Can we afford a customer service initiative? PMID- 11009815 TI - Strong service, strong brand help systems establish respected, recognized name. PMID- 11009816 TI - Hospitals are beginning to make creative use of excess capacity. PMID- 11009817 TI - Better HMO rates bring good news, bad news. PMID- 11009819 TI - Learning from our customers. PMID- 11009818 TI - CEO of big California capitated physicians group isn't complaining about managed care. PMID- 11009820 TI - Beyond managed care. PMID- 11009821 TI - The tug-of-war over Medicare funding. PMID- 11009822 TI - "Best-in-class": an interview with John T. Bigalke. AB - John T. Bigalke, FHFMA, MBA, CPA, is national director, healthcare assurance & advisory services, Deloitte & Touche LLP. Bigalke began his career in health care in 1977 as a staff auditor and consultant with a Big Five accounting firm. In 1983, he moved to another Big Five firm, where he spent 15 years in leadership positions, including vice chairman of the healthcare practice, before moving to Deloitte & Touche in 1998. A member of HFMA since 1980, he served on HFMA's National Board of Directors from 1997 to 2000. PMID- 11009823 TI - Integration strategies in transition: an interview with Russell C. Coile, Jr. AB - Russell C. Coile, Jr., is a nationally recognized futurist and speaker specializing in healthcare business strategy and market forecasting, and vice president and national strategy consultant for Southfield, Michigan-based Superior Consultant Company, Inc. Coile also has written seven books dealing with the future of health care, including New Century Healthcare: Strategies for Providers, Purchasers, and Plans, which was published this year. Coile spoke with HFM to share his views regarding the future of IDSs and how these organizations may need to modify their strategies to ensure their long-term financial success. PMID- 11009824 TI - Clinicians hold the key to profitable managed care contracting. AB - A managed care contract's effect on the financial health of an organization is related in part to utilization and quality management issues. Physicians and other clinicians who are involved in case management can illuminate how these issues, in relation to the contract terms, can affect the organization's bottom line. Clinicians should be part of a contract-negotiating team that also includes financial professionals and contract negotiators, in addition to legal counsel. PMID- 11009825 TI - Full amnesty could encourage provider self-disclosure. AB - The Federal government's current policy toward healthcare providers that voluntarily disclose improprieties has been ineffective because it offers no guarantee of immunity from prosecution. To be successful, a self-disclosure program must offer real incentives to providers to come forward. The government's self-disclosure programs with respect to tax, environmental, and antitrust laws provide models for an effective amnesty program. The success of these three programs, and particularly of the antitrust program, suggests that healthcare providers would be encouraged to come forward and disclose improprieties if, under certain specific conditions, the OIG and DOJ offered a guarantee of full amnesty to the entity and its officers, directors, and employees. PMID- 11009826 TI - Cost-effective compliance for the small provider. AB - Small provider organizations often have limited resources with which to implement a compliance program. The Office of Inspector General (OIG) has indicated that a small organization may take a less formal approach to compliance and develop a compliance program that fits its budget and resources. But small providers should take seriously the need for a compliance program despite the OIG's greater latitude toward small organizations. Whatever the size of their budget, small providers should emphasize due diligence and good-faith approaches to compliance to demonstrate to investigators and employees that they have made compliance a priority. These efforts can encourage the OIG to look more favorably upon the organization in the event of a compliance breach and also can improve the organization's financial performance by improving overall management of the organization. PMID- 11009827 TI - APCs challenge hospital EDs and outpatient clinics. AB - As the uneasily awaited Medicare outpatient prospective payment system becomes reality July 1, hospital emergency department and outpatient clinics face an even greater need for accurate coding models and systems, as well as vigorous cost reduction strategies. By the nature of the services they provide, such facilities will find securing adequate payment for appropriate, quality care to be a complex and challenging task. Those organizations that have prepared effectively, however, may benefit under the prospective payment system. PMID- 11009828 TI - Quality vs. costs? A survey of healthcare purchasing habits and concerns. AB - The results of the Fifth Annual Survey Report on Purchasing Value in Health Care show that to help control rising healthcare costs, most employers are offering employees more choice when it makes sense but fewer plans when they can strike a better bargain. Providers' favored tactics to reduce costs include reducing waste, reducing or eliminating certain services, increasing fees for some services, and improving medical management and information systems capabilities. Health plans overwhelmingly are focusing on increasing premiums to employers; they also are attempting to improve medical management and information systems capabilities. Many employers, health plans, and providers want to make their current arrangements work better. Improving the quality of customer service to purchasers and their employees is a top priority with both health plans and providers. PMID- 11009829 TI - PRRB issues new appeals hearing instructions. PMID- 11009830 TI - Automated compliance checker helps ensure billing accuracy. PMID- 11009831 TI - Meeting the challenges of the outpatient PPS. PMID- 11009832 TI - Investors can benefit from charitable giving. PMID- 11009833 TI - Assessment of biotechnology drugs: what are the issues? PMID- 11009834 TI - Pa. sues former AHERF execs. PMID- 11009835 TI - Mass. fights Harvard University for name. PMID- 11009836 TI - Whistleblowers make less off Medicaid. PMID- 11009837 TI - Tenn. systems forgo joint custody. PMID- 11009838 TI - Columbia streamlining becomes clearer. PMID- 11009839 TI - A two-front battle. Hospitals fight error reporting as they seek higher payments. PMID- 11009840 TI - Pa. weighs rules on not-for-profit deals. PMID- 11009841 TI - Forcing the issue. Californians set to use their electoral clout to place healthcare on the presidential front burner. PMID- 11009842 TI - Voting for a single-payer plan. Washington state coalition working to put universal-coverage initiative on the November ballot. PMID- 11009843 TI - On a quality roll. Michigan hospital wins NCQHC's top award for being 'ahead of the pack in many ways'. PMID- 11009844 TI - AHA moves on medical errors. As one hospital calls IOM report 'bang-on right,' association begins a data quest. PMID- 11009845 TI - Waving goodbye to healthcare. Execs are being driven away; it may be time to take a break before the industry breaks you. PMID- 11009846 TI - Organized doctors. Unionization of physicians a small but significant force as relationships with hospitals change. AB - The number of unionized physicians is still small, but those doctors are becoming a force to be reckoned with as organizing continues to grow and the fledgling union formed by the American Medical Association becomes more established. "The state of the unions in healthcare is excellent," says Barry Liebowitz, M.D., (left) president of the Doctors Council, which represents 16,000 physicians nationally. PMID- 11009847 TI - Five-hospital system comes undone. PMID- 11009848 TI - Indianapolis casts wider net in healthcare sprawl. System consolidation fuels growth; costs remain high. PMID- 11009849 TI - Wanted: more data from not-for-profits. Investors seek more-frequent reports on finances; organizations divided over best practices. PMID- 11009851 TI - Medicare reform plans take blows. PMID- 11009850 TI - CHA taking a close look at itself. Task force formed to address internal tensions resulting from association's revamp. PMID- 11009852 TI - Building health promotion into the national agenda: Washington, DC, February 14 16, 2001; a call to action. PMID- 11009853 TI - Effects of leadership style and group dynamics on enjoyment of physical activity. AB - PURPOSE: The primary purpose of this study was to examine the independent and combined effects of leadership style and group dynamics on the enjoyment of physical activity. DESIGN: A completely randomized 2 x 2 factorial design was used in which the manipulation of "leadership style" (socially enriched vs. bland) was crossed with a manipulation of "group dynamics" (socially enriched vs. bland). SETTING: The study was conducted in an aerobics studio on a university campus. SUBJECTS: The sample included 48 male and 42 female undergraduate students who were moderately active. INTERVENTION: Each participant was involved in a single session of step aerobics. A female graduate student provided either an enriched or bland series of interactions to manipulate leadership style, and a trained group of planted undergraduates was used to promote either an enriched or bland group environment. MEASURES: The outcome measures of interest were enjoyment and the probability of engaging in a similar activity in the future. RESULTS: Participants in the enriched leadership style plus enriched group dynamics condition reported higher enjoyment than did participants in the other three conditions. On average, the level of enjoyment was 22.07% higher in this condition than in the other three conditions (p < .001). The probability of future involvement was 13.93% higher for participants in the enriched group environment, irrespective of leadership style (p < .03). CONCLUSIONS: Enjoyment during physical activity is optimized when a positive and supportive leadership style is coupled with an enriched and supportive group environment. Future research is required to extend these findings to other activities and populations. PMID- 11009854 TI - The impact of the transcendental meditation program on government payments to physicians in Quebec: an update. AB - PURPOSE: To determine whether practice of the Transcendental Meditation (TM) technique can affect medical expenses. DESIGN: The evaluation was a quasi experimental, longitudinal, cost-minimization study. SETTING: Province of Quebec, Canada. SUBJECTS: This study involved 1418 Quebec health insurance enrollees who practiced the TM technique compared with 1418 subjects who were randomly selected from enrollees of the same age, sex, and region. TM subjects had chosen to begin the technique prior to learning about and choosing to enter the study. MEASURES: This 14-year, pre- and postintervention study retrospectively assessed government payments to physicians for treating the TM and comparison groups. Other medical expense data for individuals were unavailable. Data were inflation-adjusted. For each subject, least squares regression slopes were calculated to estimate pre- and postintervention annual rates of change in payments. We compared the groups' means and 1%, 5%, and 10% trimmed means (robust estimators) of the slopes. RESULTS: Before starting meditation, the yearly rate of increase in payments between groups was not significantly different (p > .17). After commencing meditation, the TM group's mean payments declined 1% to 2% annually. The comparison group's payments increased up to 11.73% annually over 6 years. There was a 13.78% mean annual difference (p = .0017). CONCLUSIONS: The results suggest that the TM technique reduced payments to physicians between 5% and 13% annually relative to comparison subjects over 6 years. Randomized studies are recommended. PMID- 11009856 TI - Low awareness of cardiovascular disease risk among low-income African-American women. AB - Focus groups were conducted with low-income African-American women in six different community settings in Northern California to assess their awareness of and concern for cardiovascular disease (CVD). These women had low awareness of the prevalence of CVD, attributed CVD to stress and low socioeconomic status, saw the media as an important source of health-related knowledge, and saw a need for more community awareness on CVD among African-American people. PMID- 11009855 TI - Hispanic women's breast and cervical cancer knowledge, attitudes, and screening behaviors. AB - PURPOSE: This study examined breast and cervical cancer knowledge, attitudes, and screening behaviors among different Hispanic populations in the United States. DESIGN: Data were collected from a random digit dial telephone survey of 8903 Hispanic adults from eight U.S. sites. Across sites, the average response rate was 83%. SETTING: Data were collected as part of the baseline assessment in a national Hispanic cancer control and prevention intervention study. SUBJECTS: Analysis was restricted to 2239 Hispanic women age 40 and older who were self identified as either Central American (n = 174), Cuban (n = 279), Mexican American (n = 1550), or Puerto Rican (n = 236). MEASURES: A bilingual survey instrument was used to solicit information on age, education, income, health insurance coverage, language use, U.S.-born status, knowledge of screening guidelines, attitudes toward cancer, and screening participation. Differences in knowledge and attitudes across Hispanic groups were assessed by either chi-square tests or analysis of variance. Logistic regression models assessed the influence of knowledge and attitudes on screening participation. RESULTS: The level of knowledge of guidelines ranged from 58.3% (Mexican Americans) to 71.8% (Cubans) for mammography, and from 41.1% (Puerto Ricans) to 55.6% (Cubans) for Pap smear among the different Hispanic populations. Attitudes also varied, with Mexican Americans and Puerto Ricans having more negative or fatalistic views of cancer than Cuban or Central Americans. Knowledge was significantly related to age, education, income, language preference, and recent screening history. Overall, attitudes were not predictive of mammography and Pap smear behavior. CONCLUSIONS: Factors related to mammography and Pap smear screening vary among the different Hispanic populations. Limitations include the cross-sectional nature of the study, self-reported measures of screening, and the limited assessment of attitudes. The data and diversity of Hispanic groups reinforce the position that ethno-regional characteristics should be clarified and addressed in cancer screening promotion efforts. The practical relationships among knowledge, attitudes, and cancer screening are not altogether clear and require further research. PMID- 11009857 TI - Tailoring and targeting a worksite health promotion program to address multiple health behaviors among blue-collar women. AB - PURPOSE: This study examined the relationship between health risks, health behaviors, stages-of-change, and behavior change priorities among blue-collar women participating in a worksite health promotion study. DESIGN: Cross sectional. SETTING: Rural manufacturing worksites in North Carolina. SUBJECTS: Participants were 859 women aged 18 and over. MEASURES: The self-administered questionnaire assessed smoking, exercise, nutrition (fat, fruits, and vegetables), and breast and cervical cancer screening behaviors. In addition, demographics, body weight, perceived health, stages-of-change, and priority for behavior change were measured. Chi-square tests and regression analysis were used to assess statistical significance. RESULTS: Overall, 28% of women smoked, 37% were completely sedentary, 82% consumed less than five daily servings of fruits and vegetables, and the majority were overweight. The dominant stage of change for each of the lifestyle behaviors was contemplation, whereas most women were in the action stage for cancer screening. When asked to prioritize the behavior they most wanted to change, the majority of women chose healthy eating and/or exercise. CONCLUSIONS: The findings suggest that blue-collar women in this study had multiple health risks and were interested in changing multiple health behaviors. Allowing women to choose the behavior(s) on which they are ready to focus may be a promising approach to tailoring interventions for this population. PMID- 11009858 TI - Closing of the Seniors Plus Social HMO in Minneapolis operated by HealthPartners (formerly Group Health, Incorporated) PMID- 11009859 TI - The application of unisex annuity tables to retirement plans. AB - Protections in pension law meant to increase women's pension income must be reevaluated in light of the increased use of defined contribution plans. One such protection is discussed: the requirement that pension plans use unisex annuity tables in calculating monthly benefits for retirees. Its usefulness is examined when applied to defined contribution plans. Legal reforms necessary to insure the use of unisex annuity tables are examined. PMID- 11009860 TI - A global perspective on social security programs for the aged. AB - The social security programs that have become widely established in countries around the globe vary greatly in their design features. The difference among the programs invites questions regarding the relative strength of their designs. One hundred and sixty-four programs are assessed on three design dimensions: benefit eligibility, qualifying eligibility criteria, and benefit provision. The analysis shows that the best-designed old-age programs can be found in Western Europe and Australasia. PMID- 11009861 TI - Coordinating health, extended care, and community support services: reforming aged care in Australia. AB - Under pressure to maximize the cost-effectiveness of programs, efforts to improve coordination have become increasingly central to the development of the broader health and welfare service delivery system in Australia in the past few years. This article reviews recent experience in two related fields: (1) the coordination of different community care services for older people and people with disabilities, funded by the Home and Community Care program; and (2) the attempt to enhance links between community and residential care services, hospitals, and other health care providers. Why coordination has emerged as such an important issue in the field of community care and, increasingly, across the entire system of what the Australian government now terms health and family services is discussed. A number of measures that have been introduced or are proposed to improve a coordination of services are briefly reviewed. These range from individualistic approaches based on information and referral, through schemes involving gatekeeping, case management and brokerage of services, to models involving the reconfiguration of organizational structures, linkages, and finances. These measures are not mutually exclusive and are increasingly likely to be applied in more complex mixed models of service coordination. It is argued that coordination at the level of direct-service provision is difficult if government policies that direct services lack coordination. PMID- 11009862 TI - Physicians and clergy as facilitators of formal services for older adults. AB - Researchers have sought to understand the determinants of the use of in-home and community-based services in order to better serve the needs of older adults. One component frequently included in formal service utilization models is the role of individuals who exert an influence on the service use process. An analysis of in depth interviews conducted with 115 older adults revealed the important facilitating role that physicians and religious leaders play in encouraging the use of these services. The sample, which included African-American and white adults age 65+ from rural and urban environments, described various ways in which these "facilitators" influence the use of formal services. These ways include: (1) supplying instrumental support either by "ordering" a particular program for or linking the elder with the program and (2) providing informational and affective support, including advising or recommending the use of a program, conveying necessary background on formal services, and legitimizing the use of formal services. Regardless of personal characteristics (such as ethnicity and residence), a majority of elders in the sample recognized the important role played by physicians, and clergy. The acknowledgement of the role played by these facilitators should be viewed as an opportunity for physicians and clergy to enhance the knowledge and appropriate use of needed formal services for elders. In addition, these findings have policy implications for the current provision of aging services. PMID- 11009863 TI - Effects of incentive size and timing on response rates to a follow-up wave of a longitudinal mailed survey. AB - Young adults who had previously participated in a longitudinal survey of youth were sent a questionnaire. They were randomly assigned to receive a $20 prepayment, a $20 postpayment, or a $25 postpayment for participation in the latest survey. Those in the large incentive condition were 7 percentage points more likely to return a survey than those in the smaller, postpayment group. Prepayment had a smaller, less reliable effect. Effects of incentive magnitude and timing were consistent at each month of the study period; only better high school grades distinguished early responders from late responders. Nonresponders had characteristics suggestive of low social conformity and were more likely than responders to be African American and male and have low SES. The discussion centers on motivations for participating in research and differences in the incentives likely to promote continued response versus initial study enrollment. PMID- 11009864 TI - Program variation in treatment outcomes among women in residential drug treatment. AB - Multilevel modeling was used to assess the program characteristics associated with treatment retention among 637 women in 16 residential drug treatment programs in the Drug Abuse Treatment Outcome Study. Women who were pregnant or had dependent children had higher rates of retention in programs in which there were higher percentages of other such women. Longer retention was associated with higher rates of posttreatment abstinence. Bivariate analyses showed that programs with higher proportions of pregnant and parenting women provided more services related to women's needs. The findings support the provision of specialized services and programs for women in order to improve outcomes of drug abuse treatment. PMID- 11009865 TI - You got a problem with that? Exploring evaluators' disagreements about ethics. AB - A random sample of American Evaluation Association (AEA) members were surveyed for their reactions to three case scenarios--informed consent, impartial reporting, and stakeholder involvement--in which an evaluator acts in a way that could be deemed ethically problematic. Significant disagreement among respondents was found for each of the scenarios, in terms of respondents' views of whether the evaluator had behaved unethically. Respondents' explanations of their judgments support the notion that general guidelines for professional behavior (such as AEA's Guiding Principles for Evaluators) can encompass sharply conflicting interpretations of how evaluators should behave in specific situations. Respondents employed in private business/consulting were less likely than those in other settings to believe that the scenarios portrayed unethical behavior by the evaluator, a finding that underscores the importance of taking contextual variables into account when analyzing evaluators' ethical perceptions. The need for increased dialogue among evaluators who represent varied perspectives on ethical issues is addressed. PMID- 11009867 TI - Technology breeds insecurity: privacy becomes a federal policy. PMID- 11009866 TI - Theory-based evaluation in practice. What do we learn? AB - Theory-based evaluation (TBE) explores the how and why of program success or failure. Advocates of TBE claim that it produces information unavailable in traditional process and outcome studies. This article examines six published papers of TBEs. It finds that the authors of the papers do not always make explicit the relation of their data to the theory of the program. Nevertheless, it was evident in one or more cases that TBE identified unnecessary program components, located intermediary changes, raised new questions, contributed to a paradigm shift, highlighted the difficulties of taking successful pilot programs to scale, and provided clarity and focus for the evaluation. Interestingly, in none of the studies was the original theory completely right. Lessons for the future of TBE are drawn. PMID- 11009869 TI - Learning the laws behind compliance. PMID- 11009868 TI - Conducting your own internal assessment. American Health Information Management Association. PMID- 11009870 TI - Exploring IT opportunities for HIM professionals. PMID- 11009871 TI - Healthcare trends--the big picture. Megatrends you need to know about. PMID- 11009872 TI - Healthcare trends. The industry response to healthcare change. New models for hospitals, health systems. PMID- 11009873 TI - Healthcare trends. The industry response to healthcare change. Long-term care: keeping the promise, today and tomorrow. PMID- 11009874 TI - Healthcare trends. The industry response to healthcare change. Home care poised for greater demand. PMID- 11009875 TI - Healthcare trends. Changing with the times. Panel discussion. PMID- 11009876 TI - Confronting ethical dilemmas on the job: an HIM professional's guide. PMID- 11009877 TI - Keeping an eye on the Joint Commission's new core measures. PMID- 11009878 TI - Practice brief. Disaster planning for health information (updated). American Health Information Management Association. PMID- 11009879 TI - The cornerstones of outstanding leadership. PMID- 11009880 TI - Reviewing diagnostic coding for mental disorders. PMID- 11009881 TI - Mental disorders pose coding challenge. PMID- 11009882 TI - Speaking up for HIM. PMID- 11009883 TI - Face lift in a jar? PMID- 11009884 TI - Different strokes. The misdiagnosis of a former President's illness shows the importance of recognizing symptoms. PMID- 11009885 TI - Technology part 1: the Internet--opportunities & threats. AB - This is the first article in a three-part series about technology and its impact on healthcare systems and the business of health care. This article explores the use and implications of Internet and Web-based technologies for physicians, managed care organizations healthcare providers. The second article will present clinical and pharmaceutical technologies. The third article will profile systems that are successfully exploiting all of these technologies. PMID- 11009886 TI - Profile: Baptist Health System, Birmingham. PMID- 11009887 TI - Health plans: not always safe. A cycle of HMO failures leave some consumers scrambling for coverage--and some without it. PMID- 11009888 TI - Calcium overload. After years of neglect, this vital mineral is showing up in everything from OJ to muffin mix. Are we getting too much of a good thing? PMID- 11009889 TI - Tapping the power of the placebo. Sugar pills are potent medicine when taken in the right spirit. Can we put them to practical use? PMID- 11009890 TI - Unemployment ramifications. A study of Deckerville Community Hospital. PMID- 11009891 TI - The evolving role of physician assistants. PMID- 11009892 TI - A house of peace. PMID- 11009893 TI - Coaching for renewal and growth. PMID- 11009894 TI - After the layoffs. PMID- 11009895 TI - Current issues in Medicaid managed care. PMID- 11009896 TI - It's all in the mix. PMID- 11009897 TI - Optimizing organization transitions: Part II. PMID- 11009899 TI - Employee lawsuits: order in the court. PMID- 11009898 TI - Help for people with head injuries. PMID- 11009900 TI - A timely turning. PMID- 11009901 TI - Do your community benefits have any impact? Part 1. PMID- 11009902 TI - Hospital staffing in rural areas. PMID- 11009903 TI - Nursing shortages: insights and solutions. PMID- 11009904 TI - The declining state of Michigan hospitals. PMID- 11009905 TI - Beyond fear. Improving documentation enhances compliance, quality and reimbursement. PMID- 11009906 TI - Tipping the scales for health care. PMID- 11009907 TI - Who makes a difference? PMID- 11009908 TI - Fighting a new war on drugs. PMID- 11009910 TI - HR budgets could use some lemon-aid. PMID- 11009909 TI - A new link. PMID- 11009911 TI - How much space do you really need? PMID- 11009912 TI - How to stop the pain. PMID- 11009913 TI - Do your community benefits have impact? Part 2. PMID- 11009914 TI - Strategic financial management. PMID- 11009915 TI - How to reduce unemployment liability. PMID- 11009916 TI - The 6 percent solution. PMID- 11009917 TI - Safe harbor rules shed new light on surgery center joint ventures. PMID- 11009918 TI - Perspectives. The wheels of policy grind slowly: top story of 1999 is Medicare Commission's decision to punt. PMID- 11009919 TI - Perspectives. Pain Act: for foes and supporters a matter of, first, do no harm. PMID- 11009920 TI - Marketplace. External reviews of plan decisions: doubts grow along with acceptance. PMID- 11009922 TI - Perspectives. HCFA coverage rule: prudent purchasing or bureaucracy run amok? PMID- 11009921 TI - Perspectives. U.S. spends a lot on health but doesn't know what it buys. PMID- 11009923 TI - Perspectives. Will we try to manage care again? This time, for drugs? PMID- 11009924 TI - Marketplace. Why payers like team assessments to gauge quality of behavioral health care. PMID- 11009925 TI - Perspectives. Home health PPS spells relief to some, but fate of sickest uncertain. PMID- 11009926 TI - Perspectives. High-tech, primary care-oriented, Finland tackles care quality, choice. PMID- 11009927 TI - Marketplace. Medicare drug coverage: trying to guess how employers would change their plans. PMID- 11009928 TI - Perspectives. Caring for the underserved: it's a tough job and nobody has to do it. PMID- 11009929 TI - Perspectives. WellPoint's new plans target uninsured, but scholars are skeptical. PMID- 11009930 TI - Use 'TRU' (total resource utilization) data to evaluate utilization, disease costs. PMID- 11009931 TI - Pennsylvania council uses risk-adjusted hospital data to improve patient care. PMID- 11009932 TI - Intensive drug therapies contribute to decline in hospital use for HIV. PMID- 11009933 TI - Most initial medical benefit denials reversed on appeal. PMID- 11009934 TI - Local medical advisory boards develop stage-specific guidelines to improve outcomes, cut costs. PMID- 11009935 TI - HMOs may not screen diabetics adequately for complications related to their disease. PMID- 11009936 TI - Hospitals to examine, improve medication safety under new plan to reduce medical errors. PMID- 11009937 TI - Updated system enhances patient identification, outcomes analysis; helps manage risk. PMID- 11009938 TI - Contract labor use in healthcare. PMID- 11009939 TI - Relations with news media: perceptions of healthcare executives. PMID- 11009940 TI - Is education negotiable? A behind-the-scenes look at resident physician unionization in a competitive business setting. PMID- 11009941 TI - Acquired Immunodeficiency Syndrome (AIDS) update, Part 1. PMID- 11009942 TI - Campaign 2000: the candidates and America's health. PMID- 11009943 TI - Readability and comprehension of the introduction to the Massachusetts Health Care Proxy. PMID- 11009944 TI - Ethical decision making in healthcare management. PMID- 11009945 TI - Decentralization and central and regional coordination of health services: the case of Switzerland. AB - As part of reforms in the health care delivery sector, decentralization is currently promoted in many countries as a means to improve performance and outcomes of national health care systems. Switzerland is an example of a country with a long-standing tradition of decentralized organization for many purposes, including health care delivery. Apart from the few aspects where the responsibility is at the federal level, it is the task of the 26 cantons to organize the provision of health services for the population of around 7 million people. This permits the system to be responsive to local priorities and interest as well as to new developments in medical and public health know-how. However, the increasing and complex difficulties of most health care delivery systems raise questions about the need for mechanisms for coordination at federal level, as well as about the equity and the effectiveness of the decentralized approach. The Swiss case shows that in a strongly decentralized system, health policy and strategy elaboration, as well as coordination mechanisms among the regional components of the system, are very hard to establish. This situation may lead to strong regional inequities in the financing of health care as well as to differences in the distribution of financial, human and material inputs into the health system. The study of the Swiss health system reveals also that, within a decentralized framework, the promotion of cost-effective interventions through a well-balanced approach towards promotional, preventive and curative services, or towards ambulatory and hospital care, is difficult to achieve, as agreements between relatively autonomous regions are difficult to obtain. Therefore, a decentralized system is not necessarily the most equitable and cost-effective way to deliver health care. PMID- 11009946 TI - An economic appraisal of alternative strategies for the delivery of MCH-FP services in urban Dhaka, Bangladesh. AB - The strategy of distributing maternal and child health and family planning (MCH FP) services at the doorsteps of the clients--through routine visits to the eligible couples by trained fieldworkers--has been instrumental in increasing the contraceptive prevalence rate (CPR), reducing fertility and attaining a considerably high immunization coverage of children and women in Bangladesh. The doorstep strategy, however, appeared to be labour-intensive and costly. With the maturity of the programme, priorities of the national MCH-FP programme have shifted to a stage that calls for more cost-effective service-delivery strategies, capable of offering a broader package of reproductive and other essential health services. The main objective of the present study was to examine the cost and effectiveness implications of the alternative strategies of delivering services from fixed sites--field-tested within an ICDDR,B operations research--in comparison to the conventional (existing) doorstep strategy. The key findings of the economic appraisal indicated that, at the end of the operations research intervention, both cost per birth averted and cost per QALY gained were lowest for the option of delivering services from static (fixed-site) clinics: US$13 and US$17 compared with the corresponding values of US$18 and US$42 for the doorstep strategy. Provision of health and family planning services from clinics- complemented with a reduced system of outreach workers to inform and target the hard-to-reach clients--was found to be the most cost-effective service-delivery alternative. PMID- 11009947 TI - Primary health care ICD--a tool for general practice research. AB - The International Classification of Diseases (ICD) has become the only diagnostic classification system of international standard that allows reliable comparisons of epidemiological data between countries, parts of a country, levels of health care systems, or different periods of time. However, for primary health care purposes it is too specific to describe problems relevant to the work of general practitioners (GPs). Tools are thus needed to 'compress' data in primary health care for educational, administrative and research purposes. The objective of this paper was to introduce a diagnosis coding system that can be used in primary health care settings for these purposes. Principal diagnoses, as assessed by medical doctors, were collected from 20,648 patient visits. After primary coding, according to the ICD on a five-digit scale, the diagnoses were grouped into larger entities, which reduced the number of diagnoses to one eighth of the original number. The ten most common original five-digit diagnoses accounted for one quarter of all consultations, whereas the ten most common diagnoses counted for one half of all consultations after compressing the data into new categories. Analysing epidemiological data in the records of primary health care by the ICD offers benefits because of the latter's wide use. These are discussed below. PMID- 11009948 TI - A question of emphasis: efficiency or equality in the provision of pharmaceuticals. AB - The reported results are part of the overall evaluation of the new drug distribution legislation that went into effect in March 1996, liberalising ownership of community pharmacies in Iceland. We addressed the following question: What impact did the legislation have on users' access to and costs of pharmaceuticals? Seven focus group discussions were conducted with pharmacy customers in different locations in May, August and October 1997. Widespread ignorance about the legislation was observed. Pharmacy customers preferred to discuss the role of physicians in 'irrational drug use' to discussing community pharmacies. A definite split was observed between urban and rural pharmacy customers; whereas definite changes were reported in the urban setting (lower prices and increased access), the rural population's perception is that it is being left out. Although the study design is not generalisable, it is clear that the equilibrium between equality and efficiency in pharmaceutical distribution in Iceland has shifted. The introduction of the free market system has increased inequality between rural and urban residents in exchange for increased efficiency. PMID- 11009949 TI - Analysing the 'new' British National Health Service. AB - The British National Health Service (NHS) has seen a proliferation of confusing and sometimes conflicting labels in recent years. It has been claimed that during the Conservative Government's period of office the 'classic' NHS was 'restructured' and then turned into the 'new' NHS. The New Labour Government of 1997 has claimed that it will deliver a 'new' NHS. If all these claims are correct, it should be possible to identify four distinct periods, with three clear 'breaks of slope' with significant changes in important indices on moving from one label to another. This paper attempts to look behind these labels by critically examining what has changed in the NHS since 1979. In simple terms, what has been 'restructured'? What is new about the 'new' NHS? Why are these changes important? It is concluded that while the NHS has certainly undergone significant changes in the past 20 years or so, the appearance of a 'new' NHS may be premature as in many areas it is possible to see the marks of continuity as well as change, and policy impact is characterised by greater incrementalism than policy rhetoric. PMID- 11009951 TI - Hospital survey process changes in 2000. PMID- 11009950 TI - Integrating outcomes into the accreditation process. PMID- 11009952 TI - Reducing solid waste in health care organizations. PMID- 11009953 TI - Sentinel event trends in wrong-site surgery. PMID- 11009954 TI - Survey process for anesthesia care standards in transition. PMID- 11009955 TI - Joint Commission mission statement revised to include patient safety. PMID- 11009956 TI - Changes to dialysis water testing requirements. PMID- 11009958 TI - Kentucky hospitals sue HHS over disproportionate-share losses tied to definition. PMID- 11009957 TI - Government releases health advisory on endoscope reprocessing. PMID- 11009959 TI - Big benefit for benefits. Antitrust concessions help Mont. system post $9 million profit. PMID- 11009960 TI - Outlook 2000. Industry turbulence may subside as Medicare pays more and merged systems look inward.... PMID- 11009961 TI - Calif. law governs not-for-profit sales. PMID- 11009962 TI - Judge rejects Calif. antitrust challenge. PMID- 11009963 TI - Native son rises. Chief executive of Albuquerque's Presbyterian Healthcare puts youthfulness to work. PMID- 11009964 TI - Cyberspace use in infancy. PMID- 11009965 TI - Back-office, online. PMID- 11009966 TI - 'Critical access' revives rural hospitals. PMID- 11009967 TI - Much at stake in new margin calculation. PMID- 11009969 TI - Healthcare costs rise in 1998. Annual percentage increase in healthcare spending since 1993 is at its highest. PMID- 11009968 TI - N.Y. reform measure fulfills most wishes. PMID- 11009970 TI - AHA: hospitals get around reporting. PMID- 11009971 TI - Flunking on charity care. PMID- 11009972 TI - New England HMOs hit hard. PMID- 11009973 TI - Mergers & acquisitions. Spinoffs, big deals dominate in '99. PMID- 11009974 TI - An HMO veteran checks out. PMID- 11009975 TI - Challenges converge on St. Louis system. PMID- 11009976 TI - AHA backpedals. Unaware of hospitals' dodging reporting requirements, it says. PMID- 11009977 TI - Miss. project aimed at resolving issues. PMID- 11009978 TI - Number of for-profits declined in 1999. PMID- 11009979 TI - Quality begins in the boardroom. PMID- 11009980 TI - 2000 Trustees of the Year. PMID- 11009981 TI - Hospitals' price hikes outpace inflation. PMID- 11009982 TI - Survey to rank systems on governance. PMID- 11009983 TI - What's a credit rating worth? PMID- 11009984 TI - Uncompensated-care spending up a little. PMID- 11009985 TI - Health system faces tough talk. PMID- 11009986 TI - Idaho docs planning to take on Columbia. PMID- 11009987 TI - One thing leads to another. Medical-errors report means money for medical outcomes research. PMID- 11009988 TI - Drugmakers shift on Medicare coverage. PMID- 11009989 TI - It may be politics over policy in 2000. AB - Thanks to election-year realities, it's likely to be politics over policy on Capitol Hill in 2000. Congress and the Clinton administration have plenty of unfinished healthcare business, but with the pressures of the campaign trail, expect a shortened legislative window of opportunity. While healthcare remains high on everyone's agenda, the likelihood of compromise remains low. PMID- 11009990 TI - Nursing homes embrace Internet. PMID- 11009991 TI - Bond volume slows from 1998 record. PMID- 11009992 TI - Whopping settlement for Fresenius. PMID- 11009993 TI - A tale of two COPAs (Certificate of Public Advantage) PMID- 11009994 TI - Online and public. Neoforma.com, first Internet-based healthcare supplier to launch IPO, soars in NASDAQ debut. PMID- 11009995 TI - Panel may support doubling of DRGs. PMID- 11009996 TI - Outpatient prices jump. Costs rose 6.7%, surpassing increase in prescription prices. PMID- 11009997 TI - Attract and retain RNs. PMID- 11009998 TI - The HIPAA lurks. Just when CIOs thought it was safe for business as usual.... AB - Just when healthcare chief information officers thought it was safe to return to business as usual, along comes the administrative simplification provisions of the Health Insurance Portability and Accountability Act of 1996. Those regulations, which will require providers to reconfigure patient records into an electronic format, will be anything but simple for hospitals, systems and insurers. PMID- 11009999 TI - Top 100 integrated healthcare networks. Intermountain tops list of best systems. PMID- 11010000 TI - Providers cut losses by selling HMOs. PMID- 11010002 TI - Hearings look at fixes for medical errors. PMID- 11010001 TI - Web wise. Leading-edge providers take Internet to its limits. PMID- 11010003 TI - Dallas PPM company files for Chapter 11. PMID- 11010004 TI - St. Louis hospital shuts doors again. PMID- 11010005 TI - West Penn to refinance AHERF debt. PMID- 11010006 TI - Who was right in Grand Rapids? PMID- 11010007 TI - CHW to sell Calif. hospital to college. PMID- 11010008 TI - For-profit chains face long-term challenges. PMID- 11010009 TI - Going back to the well. AHA kicks off campaign for $25 billion in new BBA rollbacks. PMID- 11010010 TI - Mass. Blues posts another strong year. PMID- 11010012 TI - Vanderbilt's sale of health plans stalls. PMID- 11010011 TI - Texas health plans change hands. PMID- 11010013 TI - IHS expansion leads to Chapter 11. PMID- 11010014 TI - Natural selection. Weaker hospitals find today's climate is a jungle. AB - Most hospital executives would probably agree that it's a jungle out there. In today's competitive market, it's survival of the fittest. Some observers say the healthcare industry's version of Darwinism might not be such a bad thing. The demise of weaker hospitals thins overcapacity and helps control costs. But others say it has gone too far, becoming an unhealthy evolution that's not just winnowing the weak but also bringing down the strong. PMID- 11010015 TI - Hospital companies come in from the cold. PMID- 11010016 TI - Wash. insurers must cover alternative care. PMID- 11010018 TI - First medical errors bill follows hearings. PMID- 11010017 TI - AHA tries makeover to thwart regulation. PMID- 11010019 TI - Beverly pays $175 million in fraud case. PMID- 11010020 TI - Merger now a total split. UCSF Stanford partners end talks on several joint services. PMID- 11010021 TI - Private tax-cut plans. Columbia, Tenet push for lower property assessments, despite community-benefit argument. AB - The nation's two largest for-profit hospital chains have adopted another cost cutting strategy: They've hired a small army of consultants and lawyers to persuade local tax assessors that many of their hospitals are overvalued, so they should pay less in real estate taxes. But the strategy carries some risk for the for-profits, which usually cite tax payments as one of their "community benefits." PMID- 11010022 TI - Budget dance renewed. Clinton, providers take positions on drugs, reimbursement. PMID- 11010023 TI - States take lead on quality accountability. PMID- 11010025 TI - Lobbies in fight over senior drug coverage. PMID- 11010024 TI - Health Care Hall of Fame 2000. PMID- 11010026 TI - Protocols reduce errors in Wash. heart program. PMID- 11010028 TI - Insurers pile on in Fresenius case. PMID- 11010027 TI - Mass. HMO's fall may rock hospital credits. PMID- 11010029 TI - NetCare may sell remaining hospitals. PMID- 11010030 TI - Charter seeks new deal in bankruptcy. PMID- 11010031 TI - CEO turnover drops to new low. PMID- 11010032 TI - Columbia looks to cut costs. PMID- 11010033 TI - Columbia stirs storm over Fla. CON. PMID- 11010034 TI - PHS to close Ohio teaching hospital. PMID- 11010035 TI - Tax increase funds hospital takeover. PMID- 11010036 TI - Digital mammography system OK'd by FDA. PMID- 11010037 TI - Device group: fees exceed limit. PMID- 11010038 TI - The great consolidator. Healtheon/WebMD keeps on buying in its bid to dominate online health information. PMID- 11010039 TI - Elbows fly in medical-errors game. PMID- 11010040 TI - Training shouldn't be first to go. Hospitals, health systems put quality, profitability at risk by slashing spending on staff education. PMID- 11010041 TI - Takingabigrisk.com. Execs chuck high-powered hospital jobs for e-venture. AB - Impatient with the industry's slow transition to quality improvement, an unusual group of former healthcare executives--including a canine deputy--is trying to establish a high-powered Internet venture from bargain-basement digs. The entrepreneurs, with visions of an IPO dancing in their heads, want to give providers and patients access to the information they need to improve health outcomes. PMID- 11010042 TI - Bond market changes for Tennessee system. PMID- 11010043 TI - Lobbying firm gets pork for hospitals. PMID- 11010044 TI - AMA in a $20 million hole. PMID- 11010046 TI - 'Catching' the fat bug. Can obesity be caused by exposure to a virus? PMID- 11010045 TI - The signature of death. Reviving the lost art of autopsy could be a boon for the living. PMID- 11010047 TI - Living with diabetes is getting easier--and a cure could be on the way. PMID- 11010048 TI - Positive on testing. But will the Olympic Games get clean this year--or ever? PMID- 11010049 TI - Taking aim at the citadels of slime. Bacterial 'cities' offer new insights into disease. PMID- 11010050 TI - Carcinoma renegades. Why do tumors spread? PMID- 11010051 TI - Backyard paintball play leads to serious eye injuries. PMID- 11010052 TI - Holey bike seats! You won't believe what cycling could do to you. PMID- 11010053 TI - When weak knees are the bane of strong women, special exercises help. PMID- 11010054 TI - Treatment of recurrent respiratory tract infections with a polyvalent bacterial lysate: results of an open, prospective, multinational study. AB - This multicenter, open study, carried out in 14 countries in Europe, Latin America, and Asia, recruited 4965 patients suffering from recurrent respiratory tract infections to investigate the safety and acceptability of the oral bacterial lysate immunomodulator LW 50020. Patients remained in the study for 4 months (two 4-week courses of LW 50020 separated by a 28-day treatment-free interval and follow-up). The incidence of all adverse events was 7.2%; that of adverse drug reactions was 0.6%. Adverse drug reactions were mild to moderate and not more frequent in the large subgroup of patients (77%) with a known history of allergies or underlying respiratory diseases; however, the incidence of adverse events in this subgroup was twofold higher than in the study population as a whole, probably indicating a generally increased vulnerability to disease. No clinically relevant changes in laboratory variables followed treatment. Comparison of the first study period (first course of LW 50020 and drug-free interval) with the second study period (second course of LW 50020 and follow-up) showed an overall reduction of at least 50% in the number, severity, and duration of respiratory tract infections, the number of antibiotic and symptomatic treatments, and the number of days absent from school or work. Tolerability and acceptability were assessed as good or very good in 99% of patients who completed the study. PMID- 11010055 TI - Use of ambulatory blood pressure monitoring to compare antihypertensive efficacy and safety of two angiotensin II receptor antagonists, losartan and valsartan. Losartan Trial Investigators. AB - The efficacy and safety of losartan and valsartan were evaluated in a multicenter, double-blind, randomized trial in patients with mild to moderate essential hypertension. Blood pressure responses to once-daily treatment with either losartan 50 mg (n = 93) or valsartan 80 mg (n = 94) for 6 weeks were assessed through measurements taken in the clinic and by 24-hour ambulatory blood pressure monitoring (ABPM). Both drugs significantly reduced clinic sitting systolic (SiSBP) and diastolic blood pressure (SiDBP) at 2, 4, and 6 weeks. Maximum reductions from baseline in SiSBP and SiDBP on 24-hour ABPM were also significant with the two treatments. The reduction in blood pressure was more consistent across patients in the losartan group, as indicated by a numerically smaller variability in change from baseline on all ABPM measures, which achieved significance at peak (P = .017) and during the day (P = .002). In addition, the numerically larger smoothness index with losartan suggested a more homogeneous antihypertensive effect throughout the 24-hour dosing interval. The antihypertensive response rate was 54% with losartan and 46% with valsartan. Three days after discontinuation of therapy, SiDBP remained below baseline in 73% of losartan and 63% of valsartan patients. Both agents were generally well tolerated. Losartan, but not valsartan, significantly decreased serum uric acid an average 0.4 mg/dL at week 6. In conclusion, once-daily losartan 50 mg and valsartan 80 mg had similar antihypertensive effects in patients with mild to moderate essential hypertension. Losartan produced a more consistent blood pressure-lowering response and significantly lowered uric acid, suggesting potentially meaningful differences between these two A II receptor antagonists. PMID- 11010056 TI - Spine fusion for discogenic low back pain: outcomes in patients treated with or without pulsed electromagnetic field stimulation. AB - Sixty-one randomly selected patients who underwent lumbar fusion surgeries for discogenic low back pain between 1987 and 1994 were retrospectively studied. All patients had failed to respond to preoperative conservative treatments. Forty-two patients received adjunctive therapy with pulsed electromagnetic field (PEMF) stimulation, and 19 patients received no electrical stimulation of any kind. Average follow-up time was 15.6 months postoperatively. Fusion succeeded in 97.6% of the PEMF group and in 52.6% of the unstimulated group (P < .001). The observed agreement between clinical and radiographic outcome was 75%. The use of PEMF stimulation enhances bony bridging in lumbar spinal fusions. Successful fusion underlies a good clinical outcome in patients with discogenic low back pain. PMID- 11010057 TI - A case of chronic adrenocortical insufficiency with iatrogenic anasarca. AB - The decrease in active hormones that characterizes chronic adrenal insufficiency results in hypovolemia. In some patients, residual adrenal function, mineralocorticoid therapy, and concomitant heart or liver failure, or both, can paradoxically provoke edema. The case report that follows describes a patient with iatrogenically induced anasarca resulting from the unhappy confluence of usually appropriate therapy and coexisting medical conditions. PMID- 11010058 TI - Managing chronic nonmalignant pain: overcoming obstacles to the use of opioids. AB - Physicians involved in cancer pain management treat thousands of patients with opioids, whose effective analgesia improves overall functioning. Side effects generally are tolerable, and treatment can be maintained with stable doses for long periods. Problems with addiction are infrequent. Many physicians, however, assume that opioids should be used only for chronic malignant pain. Research and clinical experience have demonstrated that opioids can safely and effectively relieve most chronic moderate to severe nonmalignant pain. Fears of addiction, disciplinary action, and adverse effects result in ineffective pain management. With current information on the use of opioids in chronic nonmalignant pain, primary care physicians can overcome these obstacles. Guidelines must clearly define the role of the primary care physician in the proper management of pain and the integration of opioid therapy. Used appropriately, opioids may represent the only source of relief for many patients. PMID- 11010059 TI - A new look at dry eye disease and its treatment. AB - This review examines the impact of moderate to severe dry eye disease on daily life and medical-resource utilization. The results suggest that current treatment paradigms can lead to unacceptable costs in both quality of life and progressive use of healthcare resources. Evidence linking this disease to T-cell-mediated inflammatory processes lays the foundation for understanding the clinical benefits of topical cyclosporine, an immunomodulatory and anti-inflammatory agent. PMID- 11010060 TI - Efficacy and safety of ketorolac tromethamine 0.5% and levocabastine 0.05%: a multicenter comparison in patients with seasonal allergic conjunctivitis. AB - This multicenter, double-masked, randomized, parallel-group study compared the efficacy and safety of ketorolac tromethamine 0.5% ophthalmic solution with levocabastine 0.05% and ketorolac tromethamine vehicle in patients with seasonal allergic conjunctivitis. One drop of ketorolac, levocabastine, or vehicle was instilled in each eye four times daily for 6 weeks. In the majority of efficacy variables, ketorolac produced the greatest improvements, followed by levocabastine and vehicle. Ketorolac was significantly more effective (P < .05) than vehicle in reducing mean itching scores, palpebral hyperemia, bulbar hyperemia, and edema. Patients treated with ketorolac reported significant improvements (P < .05) in their ability to sleep and to concentrate on work, compared with those who received vehicle. No significant differences were noted among the treatment groups in safety or tolerability. Ketorolac tromethamine 0.5% ophthalmic solution instilled four times daily is effective and safe in reducing the signs and symptoms of seasonal allergic conjunctivitis. PMID- 11010061 TI - Secondary health conditions among middle-aged individuals with chronic physical disabilities: implications for unmet needs for services. AB - Data from the Aging with Disability (AwD) Study are used to examine variations in the types and frequency of secondary conditions experienced by 301 middle-aged individuals living with the effects of three disabling conditions: polio (n = 124), rheumatoid arthritis (RA) (n = 103), and stroke (n = 75). All respondents were randomly selected from a county rehabilitation hospital or a community-based subject pool. Secondary conditions are operationalized as (1) the number of new health problems diagnosed or treated since the onset of the primary disability and (2) the amount of change/decline in basic and instrumental daily activities since a previous reference period in the disability trajectory. Also analyzed are changes in use of assistive devices and unmet needs for services. Differences in secondary conditions are examined within the AwD sample by impairment group and between samples by comparing AwD rates to national estimates for the same cohort. Results reveal significant differences in the types of new health problems reported by persons living with polio, RA, and stroke and document marked disparities, or accelerated aging, between disabled and nondisabled adults. Findings are discussed in terms of the changing health care needs of persons aging with disability and the importance of improving access to preventive services, ongoing rehabilitation, and assistive technology. PMID- 11010062 TI - Functional changes in persons aging with spinal cord injury. AB - Declines in general physical function and the effect of these changes on activities of daily living and needs for assistance were assessed in 150 individuals with spinal cord injury (SCI). The sample consisted of outpatients returning for follow-up at the spinal cord injury clinic at Rancho Los Amigos National Rehabilitation Center in Downey, CA. The average duration of injury of the sample was 13 years (range 1-37), and average age was 38 years (range 18-64). Twenty-four percent of those sampled experienced a decline or change in their physical function within the last 5 years. Individuals experiencing declines were significantly older than those without changes, averaging 45 years versus 36 years. The group with change also had a longer duration of injury, averaging 18 years versus 11 years compared to the group reporting no declines. Fatigue was the most frequently reported problem, followed by pain and weakness. Over half of the group with changes required additional assistance with activities of daily living (ADLs). Family members were the primary helpers for both ADLs and instrumental activities of daily living (IADLs) for those persons experiencing changes. The use of assistive technology, primarily equipment for bathing, toileting, and low technology devices, increased with time. The increased vulnerability to loss of function in persons with advancing age and duration of injury in this population with SCI suggests the need for early preventative measures, routine assessments to detect changes, and access to health care and supportive services to alleviate or minimize the effect of these changes. PMID- 11010063 TI - Expectations of health, independence, and quality of life among aging spinal cord injured adults. AB - While our understanding of aging and mortality in spinal cord injury is evolving, precise estimates are still not available to assist people with spinal cord injuries in knowing what to expect as they grow older. The work of a number of authors suggests that the life expectancy of those with spinal cord injuries is improving. However, little is known about the conditions in which individuals may expect to spend their remaining years of life. This study used information from a 50-year database on spinal cord injury, in combination with national mortality statistics and new survey information, to estimate the number of remaining years that individuals could expect to spend in a variety of states of health, independence, and quality of life. The study showed that expectations of health are similar to those found in the general population. Regardless of total life expectancy, individuals could expect to spend about six of their remaining years in poor health, presumably near the end of life. Expectations of independence varied depending on lesion level. Those with paraplegia became less independent over time, moving from complete to modified independence. Those with quadriplegia appeared to have a greater expectation of independence over time; however, in actuality, only those who were most independent survived to report outcomes at older ages. Expectations of quality of life remained high until the fifth decade postinjury. PMID- 11010064 TI - Work problems and accommodations reported by persons who are postpolio or have a spinal cord injury. AB - A number of studies have documented early functional declines in persons with a disability. The purpose of this study was to document (1) whether employees who are aging with their disability have experienced new work problems as a consequence of functional declines and (2) whether their work problems are being accommodated adequately. Ninety-six individuals with a disability (50 who are postpolio and 46 who had a spinal cord injury) were interviewed by phone. Each had worked at least 5 years postonset and was either currently working or unemployed for less than 5 years at the time of the interview. Forty-nine of the 50 persons who are postpolio reported they had experienced functional declines in recent years, and 41 of the 50 rated the severity of their disability greater than it was when they first began working. As a result of the functional declines they had experienced, most (90.9%) of their work problems were new and would not have been significant problems for them when they first began working. The situation was very different for the group with spinal cord injuries. Only a few members of that group had experienced functional declines that were causing new problems at work. A total of 480 work problems were reported by study participants. Three out of every eight problems did not have an accommodation satisfactory to the employee. The primary reason why a satisfactory solution was not provided was that no accommodation had been identified. Employers were generally supportive of the employee's need for accommodation; they paid for 59.1% of the accommodations that had a cost and refused to provide an accommodation for only 18 of the 480 problems. PMID- 11010065 TI - Quality of life while aging with a disability. AB - Being able to develop or maintain a positive, or high, quality of life (QOL) after a disability is one of the most important outcomes from rehabilitation. Negative, or low, QOL may also occur, especially when people return to the community. Furthermore, as people with disability age, many experience changes in their health and functioning that challenge their hard-won QOL. This article describes research on QOL among persons who are aging with a disability. Both positive and negative dimensions of QOL were studied. Being able to maintain community activities was significantly related to QOL. Current age and age at onset of disability played a nonsignificant role on negative QOL but were significantly related to positive QOL. Assistive technology can play a significant role in helping individuals who are aging with a disability to maintain valued activities and experiences. PMID- 11010066 TI - Field of assistive and rehabilitation technology. PMID- 11010067 TI - Introduction. The importance of aging with a disability to assistive technology. PMID- 11010068 TI - Aging with a disability: views from the National Institute on Disability and Rehabilitation Research. AB - As I move toward the completion of my term as Director of the National Institute on Disability and Rehabilitation Research (NIDRR), I see many examples of renewal in our field. For example, disability studies and rehabilitation science are emerging new areas of academic specialization, and we will all be involved in shaping the future of these endeavors. The field is recapturing its identity and relevance locally and internationally. Also, I see a considerable number of challenges that range from acute care in medical rehabilitation to universal design in engineering to overall technology policy. I see a challenge in moving research to practice, a challenge in funding, a challenge in service delivery locations, and a challenge in defining the role of the professional in order to build capacity for the future. I believe that the research field ignores practice at its own peril, including practice in older fields such as vocational rehabilitation and in newer fields such as cognitive science and access engineering. Practice must be determined by today's needs and those we foresee for tomorrow. For the NIDRR family, the Long-Range Plan provides a beacon to direct us into the new millennium. PMID- 11010069 TI - Metabolic and endocrine changes in persons aging with spinal cord injury. AB - Persons with spinal cord injury (SCI) have secondary medical disabilities that impair their ability to function. With paralysis, dramatic deleterious changes in body composition occur acutely with further adverse changes ensuing with increasing duration of injury. Lean mass, composed of skeletal muscle and bone, is lost and adiposity is relatively increased. The body composition changes may be further exacerbated by associated reductions in anabolic hormones, testosterone, and growth hormone. Individuals with SCI also have decreased levels of activity. These body composition and activity changes are associated with insulin resistance, disorders in carbohydrate and lipid metabolism, and may be associated with premature cardiovascular disease. Although limited information is available, upper body exercise and cycle ergometry of the lower extremities by functional electrical stimulation (FES) have been reported to have a salutary effect on these body composition and metabolic sequelae of paralysis. Perhaps other innovative, externally mediated forms of active exercise of the paralyzed extremities will result in an increased functional capacity, metabolic improvement, and reduction of atherosclerotic vascular disease. PMID- 11010070 TI - Medical problems in adults with cerebral palsy: case examples. AB - The United Cerebral Palsy Association estimates that there are approximately one half million adults with cerebral palsy in the United States. The number may be growing due to advances in medical care and the increased life expectancy of adults in general. Little has been published regarding their medical issues and rehabilitation needs. What little has been published has indicated almost a total neglect of specialty care and preventive medical services. Five case examples are presented to help illustrate particular medical and surgical problems not identified by either pediatric or adult healthcare providers alone. Each individual shows improvement with comprehensive team-oriented specialty care. Further study is encouraged. PMID- 11010071 TI - The family resemblance metaphor: some unfinished business of interpretive inquiry. AB - The rapidly expanding discipline of interpretive inquiry, especially in its narrative analysis form, has not been fully cognizant of certain crucial epistemological and methodological assumptions that form the ultimate basis of its purpose. Even after abandoning traditional positivist views, the related disciplines within the human sciences that are engaged in interpretive inquiry have still not discovered the core implicit assumptions that militate against a full acceptance of this form of inquiry. This article outlines the locus of these implicit assumptions and then argues that the legitimacy of these enterprises must be grounded in a well-known but heretofore undiscovered perspective, namely, Wittgenstein's notion of a family resemblance. It is argued that this metaphoric phrase is the key to unlocking the real and unique nature of narrative analysis. PMID- 11010072 TI - The church family and kin: an older rural black woman's support network and preferences for care providers. AB - Although kin and church are considered premier support sources for rural elders, few scholars have undertaken descriptive studies to explore the nature of rural Black elders' support networks and their preferences for in-home service providers. In the case study described in this article, methods of support network analysis and descriptive phenomenology were used to analyze data from five lengthy, open-ended interviews with a 94-year-old rural Black woman. The various groups and individuals of her network are labeled in her words, the network's supportive functions are described, and preferences for providers are noted. In addition, the varying structures of her home care experience with the support network members are described. Her attempts to voice and exercise her preferences for in-home service providers are explained in terms of two contrasting processes: preference uptake and preference suppression. Based on these findings, implications for appraising the appropriateness of rural elders' in-home services are discussed. PMID- 11010073 TI - Cross-cultural comparison of health perceptions, concerns, and coping strategies among Asian and Pacific Islander American elders. AB - This article compares the health perceptions, concerns, and coping strategies among elders in two Asian and Pacific Islander American communities. The qualitative comparison is conducted between elder Chamorros of Guam and elder Chinese of the United States, using a focused ethnography for data collection and the grounded theory method for cross-cultural analysis. The health perceptions of these two communities are manifest in two themes: (a) a sense of holism among body, mind, and spirit and (b) an orientation toward others. The health concerns of these two communities include universal experiences of aging, structural elements within the American health care system, and cultural changes impacting health. A significant coping strategy is adaptation to changes while maintaining continuity. Finally, implications for the delivery of culturally appropriate health care services to Asian and Pacific Islander American elders are discussed. PMID- 11010074 TI - Physician views on practicing professionalism in the corporate age. AB - Arnold Relman argues that medical education does not prepare students and residents to practice their profession in today's corporate health care system. Corporate health care administrators agree: Physicians enter the workforce unskilled in contract negotiation, evidence-based medicine, navigating bureaucratic systems, and so forth. What about practicing physicians? Do they agree as well? According to this study, they do. Feeling like decentered double agents and unprepared, physicians find themselves professionally lost, struggling to balance issues of cost and care and expressing lots of negativity toward the cultures of medicine and managed care. However, physicians are resilient. A group of physicians, who may be called proactive, are meeting the professional demands of corporate health care by becoming sophisticated about its bureaucratic organization and the ways in which their professional and personal commitments fit within the system. Following the lead of proactive physicians, the authors support Relman's thesis and education for both students and physicians requires a major overhaul. PMID- 11010075 TI - Visualism in community nursing: implications for telephone work with service users. AB - This article explores the potential effects of visualism (a prejudice in favor of the seen) on the perceived legitimacy of telephone work in community nursing. It discusses data from an exploratory study on telephone work, which used guided interviews with a purposive sample of 14 community nurses. Interviewees expressed both positive and negative ideas about telephone work and had particular concerns about the assessment of people, context, problems, and comprehension by telephone. These issues are discussed with reference to publications on visualism, metaphor, communication, and nursing work. The authors conclude that an awareness of the possible effects of visualism on ideas and practice could lead to improvements in both communication and assessment, whether carried out by telephone or face to face. PMID- 11010076 TI - Warnings about vulnerability in clients with diabetes and hypertension. AB - The purpose of the grounded theory study discussed in this article was to discover and explicate the basic social problem in clients with diabetes and hypertension that affects their adherence to health care directives. In-depth interviews with 21 clients with both diabetes and hypertension and 3 health care providers who care for such clients were concurrently conducted, coded, and analyzed according to the grounded theory method. Clients with diabetes and hypertension described being bombarded repeatedly with warnings about their vulnerability. The warnings were both external (coming from health care providers, family, and friends) and internal (coming from within themselves). Internal warnings were discovered to be far more influential in affecting adherence. This discovery can be used to shape health care providers' social interactions and treatment plan strategies with clients with diabetes and hypertension. PMID- 11010077 TI - Researching illness and injury: methodological considerations. AB - Circumstances surrounding the physical condition of the critically ill, the injured, and the dying make the conduct of qualitative research particularly difficult. Assumptions embedded in qualitative research are challenged or no longer apply: As sick people, participants are unfamiliar with their everyday worlds, and they are often incapable of describing their conditions and perceptions, so that researchers have difficulty obtaining data to comprehend, interpret, and generally conduct their research. Methodological problems extending from the participants' condition include the lack of everyday language to describe their experiences, the instability of the participants' reality, and the instability of the self. When researching participants who are sick, these methodological problems result in decisions about the timing of data collection, challenges to validity and reliability, and debates about who should be conducting this research. PMID- 11010078 TI - The unsolicited diary as a qualitative research tool for advanced research capacity in the field of health and illness. AB - In this article, the place of diary method is examined in the context of qualitative tools. Although much neglected in qualitative methodology, there is considerable support for its use, and it is argued that unsolicited diary analysis as a qualitative tool has value as a social research method. Although it is commissioned or solicited diaries that are increasingly popular in health research, different kinds of diary usage are examined together with certain possible biases and weaknesses. Although framework analysis was originally developed for analyzing interview data, it is seen as an appropriate qualitative tool for the analysis of unsolicited diaries. The analysis of an unsolicited diary account of a patient suffering from cancer of the larynx is explored as a potential data source. PMID- 11010079 TI - The utility of sentinel node biopsy in managing melanoma. PMID- 11010080 TI - Acne therapy. An expanding arsenal offers relief and cosmesis. PMID- 11010081 TI - Managing minor closed head injury in children. PMID- 11010082 TI - Brain injuries in athletics. Assessment and management to prevent a catastrophic outcome. PMID- 11010083 TI - Warts and children: can they be separated? PMID- 11010084 TI - IHC eyes E-health Code of Ethics. PMID- 11010085 TI - Embarking on a compliance career. PMID- 11010086 TI - "Dog tags" make data storage easier. PMID- 11010087 TI - Enhance your organization's awareness of HIPAA. PMID- 11010089 TI - Compliance across the continuum: what works. Compliance on a shoestring: one hospice's approach. PMID- 11010088 TI - Compliance across the continuum: what works. New directions for long-term care. PMID- 11010090 TI - Compliance across the continuum: what works. Communication is key to home health compliance. PMID- 11010091 TI - Comparing the OIG guidances. PMID- 11010092 TI - Compliance efforts lead to process improvements. AB - After Medicare regulations and the accompanying paperwork threatened to overwhelm an outpatient rehabilitation center, staff took steps to simplify the recertification process. The result was improved efficiency plus increased commitment to the electronic medical record. Here's how they did it. PMID- 11010093 TI - Keys to auditing hospital and professional fee coding. AB - A good audit program can save a provider from sanctions or fines. Whether you work on the hospital or physician side, here are some tips for better auditing practices. PMID- 11010094 TI - Four critical steps to choosing an auditor. PMID- 11010095 TI - Who answers the medical necessity question? PMID- 11010096 TI - Ensuring safety and security in a home office. PMID- 11010097 TI - How to write an effective resume. PMID- 11010098 TI - Practice brief. Issue: storage media for health information. AHIMA. American Health Information Management Association. PMID- 11010099 TI - How to code symptoms and definitive diagnoses. PMID- 11010100 TI - Understanding the complexities of coding for mental disorders. PMID- 11010101 TI - Outpatient prospective payment becomes a reality. PMID- 11010102 TI - Rising to the task. PMID- 11010103 TI - Understanding HIPAA's transaction and code set standards. American Health Information Management Association. PMID- 11010104 TI - Documentation errors result in Medicare overpayment. PMID- 11010105 TI - Three steps to increasing employee information security awareness. PMID- 11010106 TI - APCs: a special report. Preparing for the outpatient prospective payment system. PMID- 11010107 TI - APCs: a special report. Making a proactive transition to APCs. PMID- 11010108 TI - Strategies for reducing medical errors: HIM's role. PMID- 11010109 TI - Dialysis settings offer diverse opportunities. PMID- 11010110 TI - Building a one-stop shop for ideas. PMID- 11010111 TI - Setting the standard in assisted living. PMID- 11010112 TI - Practice brief. Letters of agreement/contracts. American Health Information Management Association. PMID- 11010113 TI - Surviving and thriving during contract negotiations. PMID- 11010114 TI - Coding for APCs: case studies. PMID- 11010115 TI - Confronting educational challenges: lead, follow, or get out of the way. PMID- 11010116 TI - Improving coding for right heart catheterization. PMID- 11010117 TI - A dot-com coder. PMID- 11010118 TI - Retirement plans, personal saving, and saving adequacy. AB - This Issue Brief addresses three questions raised by recent trends in personal saving: How are national savings measured and what is the meaning of the trends in measured personal saving rates, given what is included and what is not included in those measures? What is the effect of retirement saving programs--in particular, 401(k) plans and individual retirement accounts (IRAs)--on personal saving levels? What are the implications of existing saving behavior for the retirement income security of today's workers? The National Income and Product Accounts (NIPA), the most commonly referenced gauge of personal saving, is a widely misunderstood measure. One could argue that a complete measure of saving would include increases in wealth through capital gains, but NIPA does not factor accrued and realized capital gains on stocks and other assets into the saving rate. By one measure, accounting for capital gains results in an aggregate personal saving rate of 33 percent--more than double the rate of four decades ago. A major policy question is the impact of tax-qualified retirement saving plans (i.e., IRAs and 401(k) plans) on personal saving rates. Empirical analysis of this issue is extremely challenging and findings have been contradictory. These programs now represent an enormous store of retirement-earmarked wealth in tax-deferred vehicles: Combined, such tax-deferred retirement accounts currently have assets of about $4 trillion. Ninety percent of IRA contributions are now the result of "rollovers" as employees leave employer plans, like 401(k) plans. While leakage from the system remains a challenge, the majority of the assets in the system can be expected to be available to fund workers' retirements. One could argue that, from a retirement income security perspective, workers in general are better off because IRA and 401(k) programs exist. Surely, many of the dollars in these programs would have been saved even without the programs; but they would not necessarily have been earmarked for retirement and been available to fund retirement expenses. As rollovers become larger, this "partnership" of employment based qualified plans and IRAs will grow even more important. The evidence indicates that many groups of American workers appear unlikely to be able to afford a retirement that maintains their current lifestyle (at least not without working more years than currently planned). Consensus does not exist on how many workers are at risk or the typical magnitude of their retirement saving shortfall. There is a consensus, however, that a substantial number of individuals are at risk. This is not surprising--despite the fact that the 70 percent of workers are saving for retirement--since relatively few workers know how much it is that they need to accumulate to fund their retirement. PMID- 11010119 TI - Employer-sponsored long-term care insurance: best practices for increasing sponsorship. AB - Behind the enthusiasm of policymakers for long-term care (LTC) insurance is the belief that increased ownership of private LTC insurance will reduce the government's future liability for financing the nation's LTC needs, currently projected by the Congressional Budget Office to increase by 2.6 percent annually between 2000 and 2040. Some observers say that sustained economic growth could keep these increased expenditures at the same share of total GDP; others argue that current federal expenditure trends will become unsustainable without large tax increases. The potential of the employer-sponsored group LTC market to stave off a national LTC financing crisis has recently started to receive popular notice in the news media. However, for the potential of the group LTC market to be realized, there must be widespread employer sponsorship of group LTC plans and significant participation levels among eligible employees in these plans. The present analysis of industry data estimates the LTC plan sponsorship rate for all U.S. employers with 10 or more employees at 0.2 percent. The sponsorship rate among large employers is significantly higher (8.7 percent). The greatest growth opportunities are projected to lie in the smaller employer market, because it is enormous and virtually untapped. Nonsponsors cite a variety of barriers to employer sponsorship of LTC plans. For many nonsponsors, the most important obstacles are the intrinsic characteristics of their work forces: employees are too young, transient, part-time, and/or low-income to be suitable for LTC insurance. For many others, lack of awareness and low priority are the primary obstacles. Because group LTC insurance has been widely available for only 10 years, many benefits managers view it as "too new and untested." Prior to the passage of the Health Insurance Portability and Accountability Act (HIPAA), in August 1996, the tax treatment of long-term care insurance premiums was unclear because Congress had not addressed the issue and the Internal Revenue Service had not issued clear guidance. In essence, HIPAA served to clarify the tax status of LTC insurance and establish product criteria for tax qualification. The interventions contained in HIPAA appear to have been insufficient to stimulate coverage growth rates that will meaningfully reduced the future burden on government financing of LTC. Although employment-based LTC insurance appears to be the best mechanism for mass expansion of coverage at affordable rates, the data suggest that employer sponsorship of LTC plans is relatively rare, especially among smaller employers, and that sponsorship rates may not dramatically increase without significant investments in employer education and new incentives. PMID- 11010120 TI - Voluntary long-term care insurance: best practices for increasing employee participation. AB - This is the second of two Issue Briefs (April and May 2000) on long-term care (LTC) insurance. The previous Issue Brief addressed the problem of increasing sponsorship, while this report addresses the issue of increasing employee participation. Participation rates in group LTC insurance plans tend to be low. A potential watershed event for the development of the employment-based group LTC market is the proposed LTC program for federal employees and retirees (a program that would have to be enacted by Congress). The perception of a successful offering to federal employees could provide an enormous boost to the group LTC insurance market. Employee communication and education are seen as critical to the success of LTC enrollments. The importance of support shown by an employer for a new LTC plan offering cannot be overstated. Unlike 401(k) plan participation trends, LTC participation rates are highest among large companies. Insurers tend to view the 40-60 age range as the primary target for group LTC insurance, and employee salary as the best predictor of LTC insurance enrollment. Higher educational levels also are associated with higher levels of LTC participation. Perceived need for LTC insurance is perhaps the biggest barrier to the purchase of LTC insurance by employees due to competing financial priorities and the fact that LTC issues are generally off the "radar screens" of younger employees. Plans with skilled nursing home and home care benefits experience higher participation rates than plans lacking these benefits. The availability of lower-cost and long duration benefit options can be an important factor in determining participation. Most sponsors have chosen to offer noncontributory (i.e., fully employee-paid) LTC plans. Employer reluctance to make contributions may be caused by HIPAA's prohibition on the inclusion of LTC insurance in cafeteria plans. One of the major advantages of group LTC plans is the availability of guaranteed issue (i.e., issuing coverage without requiring evidence of insurability) for employees, which is not available in the individual LTC market. It is easy for enrollment to be derailed by the presence of any of a number of harmful conditions, such as employer-sponsors who distance themselves from the offer, ineffective communications, or difficult enrollment processes. Achieving consistently strong levels of participation in LTC plans will require employer-sponsors and their insurance carriers to form strong partnerships, with worker participation as their primary stated goal. PMID- 11010121 TI - Personal account retirement plans: an analysis of the survey of consumers finances. AB - This report is based on data from the Federal Reserve Board's triennial Survey of Consumer Financies (SCF), which provides the most comprehensive data available on the wealth of American households. The most recent SCF data are for 1998, and this report tracks information from the 1992, 1995, and 1998 surveys. The percentage of families with a participant in a pension plan from a current job increased from 38.8 percent to 41.0 percent over this six-year period. If one focuses exclusively on those families with a worker and in which the head is under age 65, the percentage for 1998 increases to 56.8 percent. The previously documented trend toward defined contribution plans was confirmed and the significance of 401(k)-type plans for those families participating in a pension plan more than doubled, from 31.6 percent in 1992 to 64.3 percent in 1998. The percentage of family heads eligible to participate in a defined contribution plan that did so increased from 73.8 percent in 1995 to 77.3 percent in 1998. Of those families choosing not to participate in a defined contribution plan, 40.3 percent were already participating in a defined benefit plan. Overall, "personal account plans" represented nearly one-half (49.5 percent) of all the financial assets for those families with either a defined contribution plan account, IRA, or Keogh, in 1998. This was a significant increase from 43.6 percent in 1992. The average total account balance in personal account plans for families with a plan in 1998 was $78,417, an increase of 54 percent in real terms over the 1992 balance of $50,914 (expressed in 1998 dollars). For families participating in a defined contribution plan, IRA, or Keogh in 1998, 52 percent of the overall average was attributed to IRA/Keogh balances (43 percent from IRAs alone), and 44 percent of the average was from account balances in defined contribution plans with the current employer. There is a marked tendency for lower-income families to have larger percentages of their total personal account plan retirement portfolio in IRAs, although this trend appears to be fading with time. The impact of rollovers on the average total account balance for all individual account balances appears to be quite large: $152,451 for those with at least one rollover, versus $78,471 for all families participating in at least one personal account plan, regardless of whether they have had a rollover. The vast majority of the rollovers would appear to be going to IRAs, as opposed to a defined contribution plan with a new employer. PMID- 11010122 TI - Physician awareness and screening for fetal alcohol syndrome. AB - This study is concerned with physicians' consistency, aggressiveness, and perception of importance in screening children for Fetal Alcohol Syndrome (FAS). Although irreversible, early intervention can improve an affected infant's survival and success in developing to his/her best potential. This study surveyed obstetricians. Pediatricians, family and general practitioners in San Antonio, Texas in 1993 and asked them to express their perceptions of the factors most important in FAS births. They also responded to questions about the rank of importance of FAS compared to other congenital disorders and indicated that alcoholism was the most influential factor in FAS births and that FAS was the third most common congenital disorder among their pediatric patients. The study further explored the physicians' clinical method to screen for FAS and their estimates of how many of their colleagues evaluate their patients for the syndrome. PMID- 11010123 TI - Principal-agent theory: a framework for improving health care reform in Tennessee. AB - Using a framework based on principal-agent theory, this study examines problems faced by managed care organizations (MCOs) and major health care providers under the state of Tennessee's current capitation-based managed care programs called TennCare. Based on agency theory, the study proposes a framework to show how an effective collaborative relationship can be forged between the state of Tennessee and participating MCOs which takes into account the major concerns of third-party health care providers. The proposed framework further enhances realization of the state's key health care reform goals which are to control the rising costs of health care delivery and to expand health care coverage to uninsured and underinsured Tennesseans. PMID- 11010124 TI - Impact of health system factors on changes in human resource and expenditures levels in OECD countries. AB - In order to gain further insight into the system factors responsible for changes in the health workforce, this study undertook an empirical examination of the determinants of the size of the health workforce and overall health expenditures across fifteen OECD countries. Specifically, using the latest release of OECD data, the analysis estimated and evaluated the effects of variables such as the proportion of female physicians and the elderly, expenditures on ambulatory care, enrollment levels in training programs, level of public financing, and per capita income on the size of the health workforce and level of health spending between 1970-1991. The findings of this study help to place the problem of the changing health workforce within the context of the complexity of health systems. It confirms any understanding of what accounts for changes in the size of the health labor force and expenditures require disentangling the effects of variables which needs to be taken into account when considering health system reforms. PMID- 11010125 TI - Political ideologies and health-oriented beliefs and behaviors: an empirical examination of strategic issues. AB - The area of health care has been called the most important political issue of the 1990s. Attitudes toward health care reform, increasing health costs, and defensive medical practices have been examined in the public press and by academicians. In addition, a substantial amount of research has been directed toward the improvement of individual personal health due to changes in personal health-related habits and behaviors. To date, there are relatively few studies which have attempted to examine the political tendencies of a nationwide sample of respondents as they relate to personal health-related beliefs and behaviors. This article explores the consumer's views on critical questions relating to health orientations and political tendencies. The results indicate a divergence between the political orientations of respondents and their beliefs and behaviors associated with health and wellness. Implications for policy-makers are discussed. PMID- 11010126 TI - The ABCs for deciding on a decision support system in the health care industry. AB - As the information superhighway becomes clustered with various organizations sharing the hope of staying solvent with the aid of technology, managers in the health care industry are more and more looking to efficient Decision Support Systems (DSS). The focus of DSS selection has often been on evaluations that measure decision performance for determining the influence and efficacy of the tool. Such evaluations, in many instances, ignore pertinent measurements for a successful selection and implementation of a DSS. In this article, the authors present a simple but important set of evaluation factors that can "make or break" a DSS implementation in the health care industry. PMID- 11010127 TI - First among (UN) equals: assessing hospital performance using data envelopment analysis. AB - This article explores the use of Data Envelopment Analysis (DEA) as a comparative analytic tool to assess the relative effectiveness of competing hospitals. Building on previous research with DEA in hospital efficiency analysis, the authors extend the earlier technical efficiency-based DEA models to include qualitative outcome measures. Data from hospitals in a regional competitive health care market in South Florida are assessed using this new DEA model. PMID- 11010128 TI - Performance appraisal: a primer for the lower level health care and rehabilitation worker. AB - This article reviews the use of performance appraisal in health care and rehabilitation organizations, particularly as such an evaluation measure could be used to examine the results achieved by managerial and supervisory level personnel. This entails a reversal of the prevailing view, that is, appraisal of lower level workers by "superiors" to a counter view as seen from those at "lower levels" looking upwards. Hierarchies, of course, are facing increasing challenges. Performance appraisal can serve the needs of the people at the lower end as well as those defined as the "management." Ultimately, performance appraisal can assure that all people in an organization will achieve benefits, not just those at the managerial level. PMID- 11010130 TI - 'Diagnostic tool' helps at-risk groups to analyze performance. PMID- 11010129 TI - Open models encourage higher satisfaction rates. PMID- 11010131 TI - Health plans still operating in the red at year-end 1998. PMID- 11010132 TI - New report offers benchmarks for routine pediatrics care. PMID- 11010133 TI - Capitation is alive and well in California, data suggest. PMID- 11010134 TI - Benefit trends work against providers managing drug risk. PMID- 11010135 TI - Assuring & improving the quality of in-home services. AB - Long-term care represents the interface between health care and social services and thus includes a multitude of services and staff that deliver long-term assistance. The range of services, consumers, and environmental circumstances under the home care umbrella creates a considerable challenge for both providers and regulators. This article examines five principles to assure high-quality care: Know your customers, listen to your customers, collect good information for decision-making, understand that the group is smarter than the individual, and realize that sub-optimization is a critical challenge facing organizations. PMID- 11010136 TI - Predictions--long-term care in the next decade. Panel discussion. PMID- 11010137 TI - Supporting aging in place & assisted living through home care. AB - This article defines assisted living, discusses the pros and cons of limited regulation, scope of service, Aging-in-Place partnerships, and how home care agencies can work with assisted-living facilities to provide care. It also examines a study on assisted living in six states that is funded by the Robert Wood Johnson Foundation. PMID- 11010138 TI - Family caregivers and home care: a declaration of interdependence. AB - Family caregivers and home care providers need each other. They must recognize their interdependence and learn to work together because millions of frail older people, disabled adults, and medically fragile children rely on family members and home care providers to help them live high-quality lives to the extent possible. PMID- 11010139 TI - Measurement: the key to success for providers, payors, and patients in long-term care. AB - The complex financing structure that supports home and community-based long-term care services has resulted in the application of different quality assurance requirements to similar services. Goals for home and community-based care are viewed differently by providers, payors, and customers. This raises the issue of how goals can be harmonized and what should be used to measure goals. Current quality assurance strategies are judge insufficient to assure the delivery of high-quality services; in part due to the state of health programs and lack of unified outcome measurement. The need for multidimensional assessment to adopt a baseline measure of health status to support allocation of services and enhance successful client outcomes has become the aim for health care providers. PMID- 11010140 TI - The new hospice compliance plan: defining and addressing risk areas. Part 2. AB - The Department of Health and Human Services' (HHS) Office of the Inspector General (OIG), has advised hospices and other health care providers to formulate effective controls to ensure compliance with federal and state statutes, rules, and regulations, and private-payor health care program requirements. This is the second of three installments that focus specifically on the 28 risk areas identified in the guidance and offer strategies for incorporating them in a hospice compliance program. This article covers the areas of Expensive Care, Documentation, Interdisciplinary Group, and Kickbacks. PMID- 11010141 TI - Critical success factors for thriving under PPS. AB - Home care agencies must work hard in the months ahead to successfully implement PPS. The new system is a fair and equitable one. Home care agencies will have to co-manage cost and quality. PPS, if implemented and administered appropriately, will benefit both the provider and the patient. Proper planning and hard work throughout the transition process will drive success. The prospective system is a reality and will become effective October 1, 2000. Agencies must start now to not only accomplish and meet the basic compliance issues, but also to develop the change in processes needed to become efficient and cost effective. PMID- 11010143 TI - Ensuring access to long-term care. PMID- 11010142 TI - Canadian study shows promising results on home care cost effectiveness. AB - Researcher Marcus Hollander, PhD, a former director of continuing care in British Columbia, is leading a national effort to compare costs of providing treatment at home to alternative settings including hospitals and nursing homes. The first study compared costs incurred by home care clients to costs incurred by similar persons in skilled nursing facilities and found home care costs were significantly less for home care. The average savings ranged from 25% to 60%. The first of 15 planned research projects on the cost effectiveness of home care to be released during 2000 offers evidence of significant savings associated with providing services in the home as compared to long-term institutions. PMID- 11010144 TI - Home care equinox. PMID- 11010145 TI - Building home care into managed long-term care: the VNS CHOICE model. AB - New approaches to home and community-centered long term care offer strategic opportunities for home health providers to thrive by providing expanded service options for the growing population of chronically ill, frail elders. The traditional expertise of home health providers makes a powerful combination with the flexibility of capitation financing to successfully meet both the needs of dually eligible older adults and government mandates to reduce overall expenditures. VNS CHOICE, a home-centered, managed long-term care program, is built on the principles and delivery system of home health care. By understanding the VNS CHOICE service delivery model and lessons learned from its first two years of operating experience, home care providers across the country may identify innovations to incorporate into their agency product lines. PMID- 11010146 TI - Want capital? Try a new approach. PMID- 11010147 TI - The hospital blue chips. PMID- 11010148 TI - Can we talk? Investors call for hospital financial feedback. PMID- 11010149 TI - Riskier business. What health care organizations must do to restore investor confidence. PMID- 11010150 TI - Making choices. Crime and punishment. PMID- 11010151 TI - The bionic century. More and more, doctors will be replacing worn-out organs with donor or manufactured body parts. PMID- 11010152 TI - Proteomics: the next frontier. PMID- 11010153 TI - The uncomfortable & the unanswerable. New technologies raise an unending list of ethical questions. PMID- 11010154 TI - The ethics of e-commerce. PMID- 11010155 TI - It doesn't come easy. A survey of hospital CEOs gives insight into collaboration. PMID- 11010156 TI - Bridging the gap. By following new rules for working with physicians, health systems can create a better health care product. PMID- 11010157 TI - Lessons in scenario planning. PMID- 11010158 TI - Health care's dark days. PMID- 11010159 TI - Confronting the R&D imperative. PMID- 11010160 TI - Web wager. Interview by D.O. Weber. PMID- 11010161 TI - Lead user: a conversation with William Coyne. PMID- 11010162 TI - We participate, therefore we are. Interview by Joe Flower and Patrice Guillaume. PMID- 11010163 TI - Integrating health care in 3D. PMID- 11010164 TI - An academic medical center reaches out. PMID- 11010165 TI - Most wired innovator awards. PMID- 11010166 TI - Destabilization: it's everywhere and all at once. PMID- 11010167 TI - Building organizational fitness. PMID- 11010168 TI - The new market makers. PMID- 11010169 TI - Making choices. Service with a snarl. PMID- 11010170 TI - Complying with outpatient PPS implementation. PMID- 11010171 TI - Turnaround, sell or shut-down: how executives can decide the future of hospital systems' business and assets. PMID- 11010172 TI - Capitation now moving to less competitive markets, long-term contracts, consultant says. PMID- 11010173 TI - MedQuist prospers as hospitals step up outsourcing of medical record transcription. PMID- 11010174 TI - Nurse shortage worse than you think, but sensitivity may help retain nurses. PMID- 11010175 TI - You can win a price war started by a competitor. PMID- 11010177 TI - The value of sponsorship. PMID- 11010176 TI - Imagine the possibilities. PMID- 11010178 TI - Never underguesstimate the financial future of Medicare. PMID- 11010179 TI - Balancing a large scale of responsibility: an interview with R. Milton Johnson. PMID- 11010180 TI - Off-balance-sheet financing can generate capital for strategic development. AB - To manage their real estate portfolios effectively and obtain funding for strategic development, IDSs should consider adopting off-balance-sheet financing strategies, such as sale-and-leaseback transactions, synthetic leases, and joint venture arrangements. Under these approaches, real estate assets are moved off of the organization's balance sheet via a partial or complete transfer of ownership to a third-party entity. The organization typically retains a satisfactory degree of control over the assets as lessee in sale-and-leaseback and synthetic-lease arrangements, or limited or minority partner in a joint venture, while freeing up cash to use for other strategic purposes. PMID- 11010181 TI - A new risk management paradigm for managed care. AB - The long-term outlook for managed care may be threatened by current capitation rate-development methods that adjust risk for age, gender, geographic area, income level, and occupation, but not health status. Not only provider payments, but also patient access may be compromised as a result. New risk-adjustment methods that categorize patients into groups based on their current health status and anticipated disease progression allow fairer distribution of the essential components of costs to deliver healthcare services to the at-risk population, and improve upon HCFA's adjusted average per capita cost (AAPCC) rates and recently implemented principal inpatient diagnostic cost group (PIP-DCG) risk-adjustment methodology. PMID- 11010182 TI - Cape: imagining the possibilities for health care and HFMA. PMID- 11010183 TI - Achieving a balance between risk and return. AB - Because risk management is costly, many health plans and providers practice risk avoidance by transferring risk to other entities. Risk-bearing healthcare organizations can improve their return on assets, but to do so they need complete information about patients' health status and the availability of effective medical treatment. To improve their return on assets, providers can use risk management strategies such as growth, designing incentives to encourage providers and health plans to reduce or eliminate unnecessary variations in resource use, and improving information about the reasons for variations in resource use and controlling those variations when possible. Providers need data to analyze why variations in resource use occur and to evaluate the efficiency of their resource use. PMID- 11010184 TI - Controlled-risk foreign investment strategy can boost yields. AB - Healthcare organizations that have invested in the U.S. stock market have enjoyed high returns in recent years. After such a performance, many investment managers see little reason to investigate overseas markets, believing that the U.S. market will continue to be profitable and economic uncertainties make overseas markets too risky. However, in 1999, markets in Europe, Australia, and the Far East outperformed the S&P 500 for the first time in five years. In addition, signs such as mounting price/earnings ratios may indicate that the U.S. stock market will be less profitable than it has been in recent years. Consequently, investment managers should revisit the idea of international investing. PMID- 11010185 TI - HCFA tightens eligibility requirements for provider-based status. AB - On October 10, 2000, HCFA will implement new eligibility standards for all off campus entities with a provider-based designation. Providers that wish to retain the provider-based status of their ambulatory care facilities need to take action to more fully integrate the ambulatory care facilities with their main facilities. In addition, the provider will need to show that 75 percent of the patient population served by the ambulatory care facility also is served by the provider's main facility. PMID- 11010186 TI - OIG offers guidance on the legality of telemedicine arrangements. AB - Telemedicine increasingly is being used to enable patients in rural areas to gain access to specialists. At the same time, concerns have arisen about the legality of certain telemedicine applications. To date, the HHS Office of Inspector General (OIG) has issued two advisory opinions stating it would not impose sanctions on two specific telemedicine arrangements, where potential violations of the antikickback statute could exist. These opinions, while applying only to the specific arrangements in question, provide physicians with clues to the OIG's thinking on various arrangements they may wish to initiate. PMID- 11010187 TI - Transitional corridors delay outpatient payment reductions. PMID- 11010188 TI - Application service providers: virtual information managers. PMID- 11010189 TI - Credentials and their impact on career advancement. PMID- 11010190 TI - Data trends. Hospital financial squeeze tightening. PMID- 11010191 TI - The high-touch financial manager. PMID- 11010192 TI - Patient-friendly billing. PMID- 11010193 TI - Should Medicare remain an entitlement? PMID- 11010194 TI - The five-word CFO job description: an interview with Lowell W. Johnson. PMID- 11010195 TI - Restructuring patient financial services for maximum effectiveness. AB - Following its formation as a regional IDS, John Muir/Mt. Diablo Health System in Walnut Creek, California, adopted an innovative approach to reorganizing its patient financial services (PFS) functions. Instead of consolidating all PFS functions into a single centralized business office, the IDS divided PFS leadership responsibilities into two areas: daily operations and technical functions. The director of daily operations oversees PFS clerical staff who are responsible for admitting functions, billing, follow-up, and related activities. The director of technical functions is responsible for implementing, uploading, and maintaining the various information systems; contract tracking and compliance; and regulatory compliance. PMID- 11010196 TI - An integral approach to compliance. AB - Financial managers of managed care organizations should structure their compliance plans to conform with requirements shaped by forces such as judicial decisions and extralegal standards for managed care operations, in addition to state and Federal laws and regulations. In addition to researching the law and training employees, an integral compliance plan should include monitoring organizational systems that can encourage or impede compliance, measuring employees' motivation to adhere to compliance standards, and modifying corporate culture to support compliance goals. Managed care organizations should empower compliance officers with a broad range of tools to assess and improve compliance efforts, or risk having organizational decisions examined externally through the lens of an onerous, dynamic system of laws and judicial decisions. PMID- 11010197 TI - Achieving superior productivity. AB - Hospitals need to analyze, develop, and implement realistic, effective productivity standards. The first step in a productivity analysis is to collect performance data for each hospital department. A workload measure should be assigned to every department. Comparing historical performance data for each department highlights problem areas. Gaining executive commitment and department manager acceptance is essential to productivity-improvement initiatives. Even in departments that experience a change in function, historical data can be used to monitor performance and determine where improvement is needed. A weighting system can be employed to capture historical data and establish a standard against which to measure future performance. PMID- 11010198 TI - Your organization should consider a cash-balance pension plan. AB - In recent years, a growing number of healthcare organizations have dropped the traditional defined-benefit pension plan and adopted cash-balance pension plans. A cash-balance pension plan generally allows employers to pay less in overall pension benefits and administration costs. A cash-balance pension plan pays benefits according to a predetermined formula based on an average of the employee's annual salary over his or her length of service. This provides recognizable benefits to younger employees but lower overall benefits to employees who have a long length of service. To assuage employees who may feel cheated out of the pension benefits they expected, employers that change to a cash-balance pension plan should consider offering higher guaranteed growth rates, advanced notification of the change to the new plan, and generous early retirement options for employees with longer lengths of service. PMID- 11010200 TI - Soft landings: dissolving a physician network. AB - Many healthcare organizations established physician networks to strengthen their competitive position in the marketplace. Many of these networks, however, have been unprofitable. As a result, some healthcare organizations are dissolving their networks. Healthcare organizations should plan the dissolution carefully and help the physicians establish an infrastructure for their new practices. By easing the physicians' transition to independent practice, healthcare organizations ensure the likelihood of a continued business relationship with the physicians in the future. PMID- 11010199 TI - The changing role of internal auditors in health care. AB - Two surveys of directors of internal auditing in health care conducted in 1990 and 1998 found that healthcare internal auditors are spending proportionately more time on management and operational improvement activities and less time on traditional financial/compliance activities. The average staff size has remained relatively constant, but salaries at all levels of experience have risen. More importantly, the tenure of healthcare internal auditors has increased significantly since 1990. The profile of the healthcare internal auditing director also has changed. The director is older, more experienced, and has held the position for twice as long as was the case in 1990. On the other hand, the director is more stressed and less satisfied with compensation. PMID- 11010201 TI - OIG clarifies emergency care requirements for providers and plans. PMID- 11010202 TI - Evaluating ASP pricing models and contract terms. PMID- 11010203 TI - Supporting organizations give donors more control. PMID- 11010204 TI - The smart approach to buying cash management services. PMID- 11010205 TI - Integrity is integral to career success. PMID- 11010206 TI - Data trends. Hospital financial indicators show little improvement. PMID- 11010207 TI - How consumers evaluate health care quality: Part III. AB - This article is the third and final article in a series which examines the way in which consumers assess information regarding the quality of health care services. The previous article focused on consumers' perceptions of health care plans and health insurance companies. This article examines the views of health care consumers regarding hospitals and doctors. PMID- 11010208 TI - Effects of direct-to-consumer advertising of pharmaceutical products on college students. AB - This study examines college and university students' responses to direct-to consumer advertising of pharmaceutical products in magazines. Four hundred seventy-one students from three institutions participated in the study that found that the majority of the students were frequent magazine readers, they noticed ads for the pharmaceutical products, and they used the advertised products. Although most students denied buying or using the products because of the ads, there was a significant correlation between number of ads seen and number of products used. PMID- 11010209 TI - Client satisfaction with substance abuse treatment. AB - A survey of 15 publicly-funded treatment agencies in Iowa was conducted to identify correlates of satisfaction with substance abuse treatment for voluntary clients. We used stratified least-squares regression analysis to identify program characteristics that were associated with greater satisfaction levels. In order to investigate satisfaction among voluntary clients, we stratified the data using three measures of client choice. These were self-reports of the extent to which the respondent felt pressured by the threat of jail, legal action, or family demands. Women tended to be more satisfied than men. Satisfaction scores for clients who indicated they were in treatment by choice were correlated with judgements about program characteristics. Phone availability, time with counselor, counselor skill, and sensitivity were associated with greater levels of satisfaction in all three models. Parking, privacy and cleanliness were significant in two out of three models. PMID- 11010210 TI - The marketing function in managed care systems. AB - As managed care spreads through the health care service industry, marketing professionals are faced with the challenge of marketing highly integrated systems. This paper explores three questions related to this development: (1) what is unique about managed care marketing, (2) how has managed care impacted health care marketing, and (3) what new strategies and trends will shape these developing markets? PMID- 11010211 TI - Low recall of local television health care news segment topics, sponsors and program names. AB - In an effort to examine the effectiveness of hospitals' sponsoring of local broadcast news health care segments, viewer recall of information presented in news reports, the program name of the reports, and the names of the hospital sponsors of those reports were analyzed. Despite the fact that telephone survey respondents said they place a higher value on televised health care information than they do for other mass media, recall was found to be generally poor. A majority of respondents could not name the topic of the televised health news report they had viewed, the program name or the program sponsor. PMID- 11010212 TI - Marketing syringe/needle exchange programs. AB - In an effort to bring the AIDS epidemic under control among the highest risk population of injecting drug users, Syringe/Needle Exchange Programs are being considered, developed and implemented. This paper looks at some of the programs in the United States in various stages of development. Also discussed will be the medical, social and political issues surrounding Needle Exchange Programs. PMID- 11010213 TI - Accuracy of information on printed over-the-counter drug advertisements. AB - Direct-to-consumer drug advertising is a useful medium for educating people and disseminating product information. Consumers make product purchase decisions based on the information gained from advertisements. If advertisements are misleading, consumers may not have adequate drug knowledge to detect this misinformation. The objective of this study was to evaluate print advertisements for over-the-counter (OTC) products. Five clinical pharmacists evaluated print advertisements appearing in three consumer periodicals. Advertisements were selected over a nine month period beginning January 1994. Accuracy of information on OTC advertisements was determined based on federal guidelines. Additionally, reviewers identified deficiencies in advertisements that may mislead consumers. According to reviewers, around 50% of advertisements lacked accurate statements. Side effects were indicated on only one advertisement. All advertisements were indicated by reviewers to be more promotional than educational. Reviewers indicated that more than 50% of advertisements lacked information essential for consumers to make an informed choice during self-medication decisions. This study indicates that OTC drug advertisements lack information necessary for consumers to make informed purchase decisions. Inaccurate information and lack of information on side effects could mislead consumers causing harmful adverse events. PMID- 11010214 TI - Customer satisfaction and consumer responsibility: toward an alternative model of medical service quality. AB - In the increasingly competitive environment of medical services and patient care, physicians feel a strong pressure for increasing efforts to improve patient satisfaction with the goal of creating a loyal patient base. These steps to promote patient satisfaction have typically involved developing new programs and services, as medical offices seek to attract and keep their patients by continually enhancing service features. While patient satisfaction is a worthy goal, this paper argues that we often make mistakes and incur expensive costs in pursuing satisfaction as an end unto itself. This paper proposes an alternative model, based on creating a doctor-patient therapeutic alliance which has the dual benefits of enhancing patient satisfaction while improving the critical personal relationship between doctors and their patients, so necessary for the delivery of optimal care. PMID- 11010215 TI - Primary health care in the reforms of the health care system: an analysis of reformation schemes of emerging and developing countries as applied to the Republic of Macedonia. PMID- 11010216 TI - Access as a managed care marketing outcomes measure. AB - As their position in the health care market diminishes, HMOs are feeling the pinch from the competition. Purchasers of health plans have many more options available today than in the past. Employers can select from single or consolidated health plans, plans offered by coalitions, or plans offered by provider systems. Following closely behind the withstanding issue of controlling costs is quality of care and customer satisfaction. The bad press surrounding managed care is making employers demand assurances that employees will receive the best quality of care their money can buy. To assist in this endeavor managed care companies are focusing more on their customers. To this end marketers use report cards to assess purchaser and enrollee satisfactions, with the hope that if they have a happy customer, s/he will be a loyal one. This paper reviews current marketing strategies of managed care companies and their level of usefulness with respect to sustaining customers and hence market share. PMID- 11010217 TI - Perceptions of care and satisfaction in assisted living facilities. AB - This study had two objectives: (1) to determine current consumers' perceptions and satisfaction with assisted-living facilities and (2) to determine current consumers' evaluation and preferences for nursing assistants. Responses from residents and family members indicated positive, but not strong, satisfaction with care. Areas identified as often unsatisfactory were; the mealtime experience, the nursing assistants, and recreational activities. Preferred qualities of nursing assistants identified by both residents and family members were genuine concern, kindness, respect, and consistent attentiveness. Residents only identified a pleasant disposition as an essential quality of nursing assistants and only family members identified knowledge regarding aging, gentle assertiveness, and commitment to staying on the job as essential qualities of nursing assistants. PMID- 11010218 TI - An analysis of the demand for regular dental health care: implications for marketing. AB - The more information a dentist has concerning factors that affect the demand or lack of demand for dental care the greater his or her capability to profile client markets. Logically the more exact the profiling of clients (potential clients) the better able the dentist is to develop a marketing program that is responsive to various market segments. In this paper the authors report findings extracted from an extensive health assessment survey which shed light on factors influencing the demand for dental services. Responses from 1934 residents of a large southeastern metropolitan area were analyzed. Statistical analyses of the data were conducted to determine whether or not a relationship existed between dental care utilization and the presence/absence of dental insurance, gender, racial/ethnic background, and household income. The potential implications these factors may have on marketing a dental practice are explored and recommendations presented. PMID- 11010219 TI - Benefit and risk information in prescription drug advertising: review of empirical studies and marketing implications. AB - As pharmaceutical companies began to advertise prescription drugs directly to consumers as well as to physicians, understanding the impact of benefit and risk information in drug advertising on physicians and consumers has become more critical. This paper reviews previous empirical studies that examined the content of benefit and risk information in drug advertising and its potential effects on physicians' subsequent prescribing behaviors. It also reviews studies that investigated how consumers process information on a drug's efficacy and side effects. Based on the findings of these studies, implications are discussed for effective marketing information development as well as for government regulation. PMID- 11010220 TI - Regulatory effects in health care: philosophies, impacts, and free markets. AB - In spite of the fact that other service sectors such as airlines, retail banking, telecommunications, railroads, trucking, securities brokerage, and insurance have experienced remarkable transformations toward more deregulated environments, the health care industry continues to be very heavily regulated. This article first examines the major theories and general schools of thought surrounding the concept of regulation in general, followed by an examination of the influence governmental regulations have had on hospitals in particular, specifically as they relate to stimulating competitiveness, efficiencies, and cost control. PMID- 11010221 TI - An open system approach to process reengineering in a healthcare operational environment. AB - The objective of this study is to examine the applicability of process reengineering in a healthcare operational environment. The intake process of a mental healthcare service delivery system is analyzed systematically to identify process-related problems. A methodology which utilizes an open system orientation coupled with process reengineering is utilized to overcome operational and patient related problems associated with the pre-reengineered intake process. The systematic redesign of the intake process resulted in performance improvements in terms of cost, quality, service and timing. PMID- 11010222 TI - Direct and indirect costs of schizophrenia in community psychiatric services in Italy. The GISIES study. Interdisciplinary Study Group on the Economic Impact of Schizophrenia. AB - BACKGROUND: The present work is a cost-of-illness (COI) study that aims at assessing total direct and indirect costs of schizophrenic patients in community psychiatric services in Italy and identifying the variables that influence costs. METHODS: A retrospective prevalence-based multi-centre COI study, was designed. Ten community mental health centres (CMHC) were involved and 100 patients were recruited. Data on patients' costs were gathered through specifically designed instruments. RESULTS: More than half total direct costs were attributed to CMHC interventions. The yearly average costs of schizophrenia per patient amounted to nearly ITL 50 million: 30% for direct costs and 70% for indirect costs. CONCLUSION: CMHCs tend to manage long-term ill patients by adopting a strong community-based system of care. Schizophrenia is correlated to loss of working days and lack of well-being. From the results of this study, it might be argued that the de-institutionalisation programme has produced 'spillovers' in terms of families' greater involvement in patients management. PMID- 11010223 TI - A qualitative study of the extent to which health gain matters when choosing between groups of patients. AB - There is considerable debate about the appropriateness of allocating health care resources on the basis of the size of the health improvement that they generate. The aim of this study was to elicit the general public's views about the extent to which health gain matters vis-a-vis other considerations. A total of 60 respondents took part in group discussions designed to enable them to raise, discuss, and reflect upon, different arguments. The qualitative data showed that many responses were being generated by factors that were not directly included in the questions, and so it is difficult to meaningfully interpret the results of other studies which have asked similar questions but which have not looked at the reasons underlying the responses. However, a clear message did come through from the data; namely, that equality of access should prevail over the maximization of benefits. However, this was subject to the outcome constraint that treatments are sufficiently effective. An important question for future research, then is 'how effective do treatments have to be for the principle of equal access to apply?' PMID- 11010224 TI - Medicare's DRG-weights in a European environment: the Spanish experience. AB - Hospital payment systems are being changed to mixed systems, composed of case-mix categories and structure indicators. The Health Care Financing Administration's Diagnosis-Related-Groups (HCFA's DRG-weights are used in Catalonia as Prospective Payment System (PPS)-instruments for hospital inpatient reimbursement. The Catalonian and Spanish health systems, however, are very different from the US health environment. The aim of this study is to determine whether the HCFA's DRG weights fit the special characteristics of a European environment. To do this, cost-based weights, determined from information from the cost accounting system of two public hospitals in Barcelona, are compared with Medicare-weights. A total of 35 262 discharges representing 12 794 million pesetas are analyzed. Medicare weights do not differ globally from cost-based-weights and the adjusted correlation weighted least squares regression between the two weight-scales is 95%. There are, however, systematic deviations in six DRG-groupings. The most important deviations are concentrated in Ambulatory Surgery categories, in DRGs in which prostheses are used, and in specialties excluded from several PPSs because of extreme variables in treatment intensity. In conclusion, Medicare weights can be used to pay hospital output in European environment but they should be adjusted to avoid systematic deviations. PMID- 11010225 TI - Leadership and the UK health service. AB - This paper explores future leadership requirements for health services in the context of relevant leadership theory and the changing environment for health services in the UK. The output of leadership research is both prolific and confusing and its applicability to health services management uncertain especially in the context of constraints on the strategic managerial behaviour and choices of public service managers. The introduction of general management to the UK NHS in the 1980s, followed by an internal market for health care in 1990 should have provided the opportunity for managers to work differently and to create personal space for leadership. However, it is not known whether sustainable , new ways of leadership working have emerged although it is reasonable to hypothesis from studies elsewhere that a number of contextual and behavioural leadership models are likely to be found in the NHS. Although management researchers have explored networking and referred to the impact of the external environment of leadership, insufficient importance has been attached to date to the impact of future trends in health services on the leadership of change in the health sector. The paper argues that in future health services leadership will require much more than traditional networking with other organizations and groups and will need to focus on developing and securing external agreement to an agenda for positive change turning the apparent constraints of the external environment, determined primarily by government policies,into opportunities. In other words, the demands of external or contextual leadership will increase forcing a stronger focus on having to achieve change through others. PMID- 11010226 TI - Five giant leaps toward integrating health care delivery and ways to drive organizations to leap or get out of the way. AB - The article argues that incremental improvement of, or "tinkering" with, health care delivery is taking too long and leaves unacceptable gaps in health care quality and service. It lists five common types of quality improvement "leaps" that have proven across studies to improve the integration of care during hospital and emergency department discharge, to bring mental health and substance abuse treatment into primary care, and to allow better management of chronic diseases. It also lists five key tasks that organizations need to accomplish in order to make such leaps and then describes four ways that consumers, employers, consumer advocates, and investors force the stubborn "bullfrogs" to take a big jump forward. PMID- 11010227 TI - Changing the paradigm: planning for ambulatory care expansion in Los Angeles County using a community-based and evidence-based model. AB - In 1998, Los Angeles County's Department of Health Services (DHS) embarked on a planning process to expand ambulatory care services for the county's 2.7 million uninsured and otherwise medically indigent residents. This planning process was novel in two ways. First, it used a quantitative, needs-based approach for resource allocation to ensure an equitable distribution of safety-net ambulatory care services across the county. Second, it used a new community-based planning paradigm that took into consideration the specific needs of each of the county's eight geographic service planning areas. Together, the evidence-based approach to planning and the community-based decision-making will ensure that DHS can more equitably provide for the needs of Los Angeles County's medically indigent residents. PMID- 11010228 TI - Reassessing the plight of physician organizations in California: the uncertain future for IPAs. AB - With numerous medical groups and individual practice associations (IPAs) in California now reporting operating losses--and many approaching financial insolvency--the question arises why physician organizations are in such a tenuous situation. One line of thinking is that the problem is attributable to the market dominance of the major health plans and their ability to impose actuarially unsound low capitation rates on professional providers. This article describes four other reasons for the current plight of physician organizations: (1) a physician-centric approach to IPA governance, (2) lack of qualified staff within key operating units, (3) management reporting that is insufficient to support utilization analysis and health plan negotiations, and (4) a highly charged political process for determining physician reimbursement. IPA survival will ultimately depend upon whether IPAs are perceived by their physician members and leaders as true business operations or just as another income source. PMID- 11010229 TI - Kaiser Permanente: integrating around a care delivery model. AB - The risk-adjusted payment model mandated for Medicare + Choice organizations by the Balanced Budget Act of 1997 has significant implications for health care organizations that offer the Medicare + Choice program. Chronic medical conditions in the ambulatory care setting will become the focus of data capture, requiring significant retooling of ambulatory care operations. Health care organizations in California currently do not have the capital available to invest in the information management tools that will be required. Private practitioners continue not to have the technology and expertise to capture the data needed to run the model. Nevertheless, the risk-adjusted payment model presents the potential to better integrate care delivery around the continuum of health care needs of the population. PMID- 11010230 TI - The rise and fall of a hospital-sponsored group practice. AB - Each physician group, network, or management services organization developed as part of an "integration" strategy over the past decade is unique. The current status of each of these entities is based on a variety of factors, including the local health care economy and environment, the sponsoring organization, and the entity's leadership. Much of the current news concerning integration initiatives is negative, and significant operating losses and the disillusionment of the participants have been reported. It is important to study the failures, however, for despite the unique factors impacting each one there are universal lessons to be learned in every case. This article chronicles the causes of one integrated group practice's breakup after five years of operation and the process undertaken to return the physicians to small independent practices. PMID- 11010231 TI - The integration movement in California's workers' compensation market. AB - Over the past two decades the California workers' compensation industry has been responding to rapidly rising medical costs. The first response was to attempt to adopt principles of managed care; the second, to increase efficiency by integrating activities, first within companies, then between companies and providers, and finally across companies providing both group health and workers' compensation. This article chronicles the integration movement, analyzes the forces driving it, and discusses how contradictory government incentives have both propelled and hindered integration. PMID- 11010233 TI - Addiction as a market failure: using rational addiction results to justify tobacco regulation. AB - Tobacco regulation efforts have been criticized by some academic economists for failing to provide adequate welfare-analytic justification. This paper attempts to address these criticisms. Unlike previous research that has discussed second hand smoke and health care financing externalities, this paper develops the logic for identifying the much larger market failures attributable to the failure of smokers to fully internalize the costs of their addictive behavior. The focus is on teen addiction as a form of "intrapersonal" externality and observed adult consumption behavior consistent with partial myopia. The importance of peer effects, in the consideration of welfare impacts, is also emphasized. PMID- 11010232 TI - The National Health Service Corps for the 21st century. AB - The National Health Service Corps (NHSC) was created in 1970 to provide primary health care clinicians for the underserved. The article includes a review of the peer-reviewed and intragovernmental literature on the NHSC program from 1971 to 1998 and also presents a current profile of the program. Despite significant increases in NHSC field strength since 1991, the 2,439 clinicians meet only 12% of the need for primary health care providers in underserved areas. While the NHSC has successfully addressed clinician diversity and retention issues, community and site development remain barriers to increasing access. Most communities in need are not ready to recruit and support clinicians. The NHSC of the next millennium must work with the neediest communities to reach the appropriate stage of readiness. Only after completing the necessary "preplacement" activities can the NHSC assist in the recruitment and placement of clinicians to increase access. PMID- 11010234 TI - On the economic foundations of CEA. Ladies and gentlemen, take your positions! AB - There are still many ongoing debates about several aspects of the methodology of economic evaluations of health care interventions. Some of the disparities in recommendations on methodological issues may be traced back to different viewpoints on cost-effectiveness analysis (CEA) in general. Two important views are the welfarist approach, which aims at embedding CEA into traditional welfare economics, and the decision maker's approach, which takes a broader and more pragmatic view on CEA. The focus in welfarism may be on utility while that of the decision maker's approach may be considered to be on health. In this paper it is examined how these two views differ and how these differences may subsequently lead to debates in methodological areas. It is indicated that embedding the practical operationalization of CEA in welfare economics seems impossible. In a strict welfarist approach it is necessary to view QALYs as being utilities, although one may question whether such an approach to QALYs is appropriate. Also, equity considerations may play an important role in cost-effectiveness analysis and these should preferably be taken into account in a way that reflects societal attitudes towards an equitable distribution of health care. These equity considerations may not always be directly related to utility or efficiency. Furthermore, both camps may prefer different methods for cost measurement in areas such as productivity costs and informal care. A better recognition of the contents and origins of controversies and disputes may enhance the clarity of discussions. PMID- 11010235 TI - On stationarity and cointegration of international health expenditure and GDP. AB - This paper examines stationarity and cointegration of health expenditure and GDP, for a sample of 21 OECD countries using data for the period 1960-1997, by applying a test battery that allows robust inference to be made on the stationarity and cointegration issue. Trend stationarity and no-cointegration are tested using new country-by-country and panel tests, not previously applied in this setting. New results for country-by-country and panel tests of non stationarity and cointegration are presented. Our unit root and trend stationarity results indicate that both health expenditure and GDP are non stationary. The no-cointegration and cointegration results indicate that health expenditure and GDP are cointegrated. PMID- 11010236 TI - Properties of actuarially fair and pay-as-you-go health insurance schemes for the elderly. An OLG model approach. AB - The aged dependency ratio or ADR is growing at a fast pace in many countries. This fact causes stress to the economy and might create conflicts of interest between young and old. In this paper the properties of different health insurance systems for the elderly are analysed within an overlapping generations (OLG) model. The properties of actuarial health insurance and different variations of pay-as-you-go (PAYG) health insurance are compared. It turns out that the welfare properties of these contracts are heavily dependent on the economy's dynamic properties. Of particular importance is the magnitude of the rate of population growth relative to the interest rate. In addition, it is shown that public health insurance is associated with an inherent externality resulting in a second-best solution. PMID- 11010237 TI - Risk segmentation: goal or problem? AB - This paper traces the evolution of economists' views about risk segmentation in health insurance markets. Originally seen as a desirable goal, risk segmentation has come to be viewed as leading to abnormal profits, wasted resources, and inefficient limitations on coverage and services. We suggest that risk segmentation may be efficient if one takes an ex post view (i.e., after consumers' risks are known). From this perspective, managed care may be a much better method for achieving risk segmentation than limitations on coverage. The most serious objection to risk segmentation is the ex ante concern that it undermines long-term insurance contracts that would protect consumers against changes in lifetime risk. PMID- 11010238 TI - An efficient employer strategy for dealing with adverse selection in multiple plan offerings: an MSA example. AB - This paper outlines a feasible employee premium contribution policy, which would reduce the inefficiency associated with adverse selection when a limited coverage insurance policy is offered alongside a more generous policy. The "efficient premium contribution" is defined and is shown to lead to an efficient allocation across plans of persons who differ by risk, but it may also redistribute against higher risks. A simulation of the additional option of a catastrophic health plan (CHP) accompanied by a medical savings account (MSA) is presented. The efficiency gains from adding the MSA/catastrophic health insurance plan (CHP) option are positive but small, and the adverse consequences for high risks under an efficient employee premium are also small. PMID- 11010239 TI - Health insurance and retirement behavior: evidence from the health and retirement survey. AB - This paper studies the role of health insurance in the retirement decisions of older workers. As policymakers consider mechanisms for how to increase access to affordable health insurance for the near elderly, considerations of the potential labor force implications of such policies will be important to consider- potentially inducing retirements just at a time when the labor force is shrinking. Using data from the 1992 and 1996 waves of the Health and Retirement Survey, this study demonstrates that access to post-retirement health insurance has a large effect on retirement. Among older male workers, those with retiree health benefit offers are 68% more likely to retire (and those with non employment based insurance are 44% more likely to retire) than their counterparts who would lose employment-based health insurance upon retirement. In addition, the study demonstrated that in retirement models, when retiree health benefits are controlled for, the effects of pension coverage are reduced, suggesting that these effects may have been overestimated in the prior literature. PMID- 11010240 TI - A note on eliciting distributive preferences for health. AB - While in theory the strength of preferences for equity in health can be expressed in an 'inequality aversion parameter', in practice, analysts would have to obtain them from people's choices. We are faced with a number of methodological problems when turning to this type of empirical research. This note investigates which types of preference could explain the choices people make when responding to equity-efficiency questions of this kind. Respondents may be heavily influenced by concerns that are not related to their equity preferences, something which may lead them to choose distributions that are not consistent with models on the equity-efficiency trade-off. Specifically, a threshold effect is identified, which could explain why some people would rather prefer to concentrate than to diffuse health gains. The second aim of this note is to offer some lessons from a survey which was designed for eliciting people's distributive preferences for health gains. PMID- 11010241 TI - How the dead help me care for the living. PMID- 11010242 TI - Easy ways to track your progress. PMID- 11010243 TI - I couldn't let the patient die--not yet. PMID- 11010244 TI - My depression has made me a better doctor. PMID- 11010245 TI - Is any malpractice settlement really confidential? PMID- 11010246 TI - Is primary care paying its way? PMID- 11010247 TI - "Internists are losers". PMID- 11010248 TI - Should we stop making a fuss over HMOs? PMID- 11010249 TI - The same old problems with FFS Medicare payments. PMID- 11010250 TI - Discounted Medicare? It's already here. PMID- 11010251 TI - Where were my dad's doctors? PMID- 11010252 TI - Scapegoating won't reduce medical errors. PMID- 11010253 TI - The patient came first--mom and dad would have to wait. PMID- 11010254 TI - The fourth pearl of internship. PMID- 11010256 TI - What a behavioral specialist could add to your practice. PMID- 11010255 TI - A doctor's worst nightmare: my own child has cancer. PMID- 11010257 TI - When I treated Grace, I also treated myself. PMID- 11010259 TI - Managed care ripoffs. Don't sign a sucker deal. PMID- 11010258 TI - Your liability when you teach residents. PMID- 11010260 TI - Managed care ripoffs. Are you being paid less than you should? PMID- 11010261 TI - Plain and simple: denying payment denies care. PMID- 11010262 TI - Turn your computer into a patient education library. PMID- 11010263 TI - Supreme Court: your office can be searched without a warrant. PMID- 11010264 TI - Hang it up at 55? No way! PMID- 11010266 TI - Do you need critical-illness insurance? PMID- 11010265 TI - What are you really good at? PMID- 11010267 TI - Are online pharmacies good for your patients--and for you? PMID- 11010268 TI - Let's audit the HMOs! PMID- 11010269 TI - From private tragedy to public activism. PMID- 11010270 TI - Let's stop subsidizing lawyers at medical meetings. PMID- 11010271 TI - What makes a physician an expert in CAM? PMID- 11010272 TI - How high-tech private eyes catch malingering plaintiffs. PMID- 11010273 TI - Patient fraud--or an invasion of privacy? PMID- 11010274 TI - "Boutique" practices: good medicine--or ethical swamp? PMID- 11010275 TI - Let's put an end to code "bungling". PMID- 11010276 TI - Redefining hope. PMID- 11010277 TI - Re-engineering your practice. Never lose a chart again! PMID- 11010279 TI - The 150 best financial advisers for doctors. PMID- 11010278 TI - How I learned to be a better boss. PMID- 11010280 TI - Living--and practicing--with hepatitis C. PMID- 11010281 TI - Our patient pulled a fast one on Medicare--and on us. PMID- 11010282 TI - How do you unload an obsolete computer? PMID- 11010283 TI - Compliance guidelines for small practices: no more excuses. PMID- 11010284 TI - Oops! Doctors' worst financial blunders. PMID- 11010285 TI - Doctors and the Web. Build your own information highway. PMID- 11010286 TI - My second specialty? It's suing lawyers. PMID- 11010287 TI - A lifelong ally for cystic fibrosis patients. PMID- 11010288 TI - Acupuncture in 2000: working its way into mainstream medicine. PMID- 11010289 TI - The job paid nothing, but the rewards were great. PMID- 11010290 TI - If they treat a doctor this way, what must patients endure? PMID- 11010291 TI - My malpractice case was a blessing in disguise. PMID- 11010292 TI - Dr. Mom--an oxymoron? PMID- 11010293 TI - Doctors who go the extra mile. Treating his far-flung patients like family. PMID- 11010294 TI - If you're so smart, doctor, why aren't you rich? PMID- 11010295 TI - Endoscopy at 35,000 feet: hand me that duct tape! PMID- 11010296 TI - Malpractice consult. When the patient doesn't fit the mold. PMID- 11010297 TI - Where's managed care headed? PMID- 11010298 TI - Will health care decide who goes to the White House? PMID- 11010299 TI - Staff raises. The purse strings won't be any looser this year. PMID- 11010300 TI - How we'll survive our next computer switchover. PMID- 11010301 TI - How to cut loose from a troublesome patient. PMID- 11010302 TI - My ethical beacons: Plato, Aristotle ... and Mr. Phillips. PMID- 11010303 TI - Who'd have thought it? Patients are putting their records online. PMID- 11010304 TI - Automated system sends reports on abnormal findings. PMID- 11010305 TI - How the Internet could wring $7.3 billion out of professional fees. PMID- 11010306 TI - MR accreditation helps to assure quality service. PMID- 11010307 TI - The Q factor. Creating a culture of quality improvement. PMID- 11010308 TI - Audits measure practice quality of mammography. PMID- 11010309 TI - Chemical characterization of guanidino compounds in serum. PMID- 11010310 TI - Alcohol use and HIV pharmacotherapy. PMID- 11010311 TI - A new pathway to product standardization. AB - As the benefits of product standardization become more evident in improved financial, managerial, and clinical outcomes, tools to make the process easier will be in demand. Once a standardization program is established, e-commerce offers tools to keep it on track. PMID- 11010312 TI - Limited usefulness of essential trace element analyses in hair. PMID- 11010313 TI - Optimizing cost and service in sterilization. AB - Controlling sterilization costs is a necessity if medical device manufacturers are to remain competitive. This article looks at ways the industry can reduce these costs and also considers the benefits to be gained by improving relationships with sterilization contractors. PMID- 11010314 TI - Electron-beam sterilization. Trends and developments. AB - The latest generation of electron-beam technology, which includes higher power accelerators fitted with X-ray conversion, is offering manufacturers a broader range of sterilizing options and driving market share for this method of sterilization. PMID- 11010315 TI - VDmax. A new method for substantiating 25 kGy. AB - Neither of the standards for the validation and routine control of sterilization of medical devices by ionizing radiation provides detailed guidance on how to substantiate a 25 kGy sterilizing dose. This article describes the basis of a new method for substantiating this dose and how the resulting procedure has been applied in practice. PMID- 11010316 TI - New high-frequency weldable polyolefin films. AB - There is an increasing desire for plastic films that can be sealed using high frequency energy. Tests on new high-frequency polyolefin film structures are reported, which compare them with the characteristics and performance of poly(vinyl chloride), ethylene-vinyl acetate and thermoplastic polyurethane films. PMID- 11010317 TI - Microsystems: applications in catheters and endoscopes. AB - Microsystems are devices that incorporate a combination of microfluidic, micromechanical, microoptical and microelectronic components. This is the second article in a series that investigates the impact the technology is likely to have on medical products during the next 10 years. This article describes how that technology is being used to construct pressure sensors, temperature sensors, microsurgical instruments, ultrasound transducers and imaging devices. PMID- 11010318 TI - Notified body recommendations. AB - This article discusses the Notified Body recommendations issued by the European Forum of Notified Bodies Medical Devices and its subcommittee, the Notified Bodies Recommendations Group. Most of these recommendations represent more detailed guidance documents than the consensus statements issued by this group, which were discussed in a previous article. Some of the recommendations form the basis of the European-level MEDDEV guidance documents. PMID- 11010319 TI - Towards global in vitro diagnostic standards, Part II. AB - The first global harmonized standards for in vitro diagnostic (IVD) products are nearing completion. Six of these standards are mandated for the IVD Directive. Part II of this two-part article continues with a report on the current status of these standards and reflects on the lessons learned so far from international standardization activity. PMID- 11010320 TI - ISO 9000:2000 revisions and their implications. AB - The planned revision of the ISO 9000 series of standards means that all medical device companies will have to revise their quality systems to align them with the new ISO 9000:2000 standards. This overview outlines the changes and the factors companies need to consider, and highlights future standards activity. PMID- 11010321 TI - Health-care reforms in Germany. AB - Uncertainty over the planned health-care reforms had a negative effect on the performance of Germany's medical device industry in 1999. With hospital financing currently at a standstill and health technology assessment on the way, this article looks at what the reforms could mean for the industry. PMID- 11010322 TI - Toward an information society for all: challenges in health telematics. AB - In the emerging information society each hospital is supposed to become a node of a complex networked information system that has to be accessible to a variety of users. These include doctors, nurses, administrators and the patients themselves, whose role is briefly described. This asks for a new approach to the implementation of the interfaces of the information system, which are used to carry out different tasks by people with different interests and computer literacy. A new approach to the implementation of such interfaces, based on the concepts of User interfaces for All, is briefly outlined, describing the scientific and technical achievements in some projects partially funded by the European Commission and the activities of the International Scientific Forum "Towards an Information Society for All" set-up to discuss and promote the relevant scientific concepts and technical issues. PMID- 11010323 TI - Education and training in the MEDICOM system. AB - MEDICOM system is a world wide telematics application for electronic commerce of medical devices. It has been designed so as to provide the health care professionals with a central Internet access to up-to-date information about medical equipment from multiple manufacturers, in a particular easy and friendly way. Moreover, the Medicom system will serve the health care professionals' requirements for high-quality information about specific products in a form of multimedia presentations and that of a secure communication channel with the community of manufacturers, especially for post marketing surveillance. The system will provide the medical staff (physicians and technicians) with demonstrations of the operation procedures and the functioning of high-tech equipment in a form of virtual models. Moreover, through the medicom system the end users of medical devices can have access to on line libraries and participate in special newsgroups. This paper discusses the architectural structure of the MEDICOM system with emphasis to its educational and training functionality. PMID- 11010324 TI - Mobile Medical Data (MOMEDA)--a Personalized Medical Information System. AB - MOMEDA is a two-years project supported by the European Commission (Telematics Applications Programme of DGXIII--Health Care Sector). The main objectives of the project are the development of a compact personal information terminal for hospital and home care environments that could be used by patients and a demonstrator that allows the consulting physician to access electronic patient record data from outside the hospital, using a hand held companion device connected to GSM network. Special attention is paid to a Personalized Medical Information System (PMIS) which will allow patients to access customized disease specific information material that will enable them to fully understand in a simple and constructive form what their medical problem is, what the planned procedures are, what lifestyle they should follow during and after their hospitalization, thus becoming more qualified partners in the recovery process. Considering the fact that in most cases informed and educated patients are usually satisfied patients when the treatment is finished, patient satisfaction can be also accomplished. The design considerations for the PMIS system are presented and the implementation is discussed. PMID- 11010325 TI - Education and training in healthcare telematics. PMID- 11010326 TI - Health informatics in the context of user ownership. AB - Of fundamental importance to the successful implementation of health information systems is the public and professional acceptability of their inclusion in the healthcare process. Surprisingly, the health care sector is embracing the informatics revolution somewhat reluctantly. Even in the areas that progress can be seen and assessed, the rate of change is erratic and inconsistent when compared across the spectrum of health care informatics. The mystery intensifies if one considers the situation in global scale: despite the diversity in approaches and models used, the reluctance perseveres. One can only suspect that there is a common, underlying, reason which slows down the process and forces the whole sector in being reluctant to accept change. PMID- 11010327 TI - Evaluation of knowledge using ExaMe program on the Internet. AB - In the paper we describe the function of the ExaMe program that serves for evaluation of students' knowledge using Internet. Evaluation is based on the knowledge base of a given course. Two types of evaluation tests are described. The fixed test is appropriate for examination of students by the teacher in computer classroom connected to Internet. By this test the evaluation is done in the limited time period and all students are tested with the same evaluation test. The automatic test is appropriate for self-evaluation and self-study by students on remote places. The student can ask the ExaMe program for the test of different difficulty levels and state the number of questions required. Finally thee first experience with the ExaMe program in practice is given. PMID- 11010328 TI - The continuum of health informatics educational efforts. PMID- 11010329 TI - Health telematics applications and user acceptance from a medical student's point of view. AB - The goal of the workshop is dual. The first is to convince health care professionals of health telematics applications' usefulness by demonstrating convincing cases and the second is to examine the practical side of the story which is education and training in health telematics. The purpose of this paper is to present the way health telematics appeal to a medical student. After all it's the medical student and the young practitioner whose lifetime practice will depend on the health telematics applications that are today under development. PMID- 11010330 TI - Thoughts about curricula in health informatics. AB - In this contribution it is argued that it is difficult to talk of medical informatics education since the groups that need education in this field are not very homogeneous. Also these groups overlap. WG 1 of IMIA is in the process of producing recommendations for curricula in medical informatics. The target groups are defined by their career type, which is categorised using three axes: discipline, stage of career and level of expertise. An example is given of the knowledge requirements for medical students. PMID- 11010331 TI - Education and training in health telematics: the critical factor of user acceptance. PMID- 11010332 TI - Education in biomedical engineering: experience from a European ERASMUS program. AB - Based on recent advances in biomedical research and the developments of new equipment and techniques, the field of Biomedical Engineering and Health Care Telematics are currently undergoing a rapid evolution characterized by an increasing degree of specialization. This, in turns, imposes new requirements in advanced education, while the changing scene at European level, introduces a major challenge for harmonization and standardization of education with a focus on meeting the emerging needs. At the same time information technologies provide new means and tools supporting the educational and training activities. An initiative for the development of a multinational advanced course in Biomedical Engineering, is implemented in the University of Patras with extended collaboration of European Universities, providing a unique case for achievement of excellency. In order to take full advantage of this potential, a Quality Assurance system has been designed and implemented over the past four years, aiming to provide the appropriate framework for mutual recognition amongst the participating institutions. Additionally, the implementation of new telematic tools is scheduled for the near future, in order to provide the Course with teleconference facilities and allow a much larger number of students to remotely attend the lectures. PMID- 11010333 TI - Design and use of clinical ontologies: curricular goals for the education of health-telematics professionals. AB - In computer science, the notion of a domain ontology--a formal specification of the concepts and of the relationships among concepts that characterize an application are a--has received considerable attention. In human-computer interaction, ontologies play a key role in defining the terms with which users and computer systems communicate. Such ontologies either implicitly or explicitly drive all dialogs between the computer and the user. In the construction of health-telematics applications, professionals need to understand how to design and apply domain ontologies to ensure effective communication with end-users. We currently are revising our training program in Medical Information Sciences at Stanford University to teach professional students in health telematics how to develop effective domain ontologies. Instruction concerning the construction and application of clinical domain ontologies should become an integral component of all health telematics curricula. PMID- 11010334 TI - Inter-university cooperation for establishing an M.Sc. course in health informatics aims at contributing to users' acceptance of health telematics applications. AB - Health informatics is a recently established and important multi-disciplinary and inter-disciplinary field that not only involves informatics but also medicine, nursing, engineering, biology and other-related subjects. A coordination of this field at a postgraduate level becomes important now in Europe where other European Community programs such as the Telematics for Health Care will require at the Fourth Framework Programme (1994-1999) and the Fifth Framework Programme (2000-2006) adequate human resources of higher potential and knowledge. A European M.Sc. course met all the above objectives. The curriculum was developed according to previous experiences in similar programmes. Recently the course has been organised on the basis of an Inter-University nature with the participation of 5 Greek Universities. The paper aims at providing a description of the new academic programme and a brief evaluation of the implementation phase. PMID- 11010335 TI - Role of the EU commission for pushing the user acceptance of the health telematics applications. PMID- 11010336 TI - User acceptance by individuals of health telematics from distributed EPRs associated with knowledge couplers. AB - The patient in a hospital bed is also a private individual that might access his or her own Electronic Patient Record (EPR) in association with additional tools that are able to show critical up-to-date knowledge about diagnoses, clinical investigations or treatments. The distributed EPRs are shown here in full production with the DOMED application of DIOGENE 2. Besides, in order to facilitate an easier understanding of the EPRs by the patient, the HON Web Site services are at his or her disposal in this respect at the same terminal. PMID- 11010337 TI - Utilising multimedia for training merchant mariners as paramedics. AB - MERMAID is a telemedicine project with global reach and a 24-hour, multilingual capability. It aspires to provide a model for the provision of health care services based on the electronic transmission of medical information, via ISDN based videoconferencing. This model is not limited to medical diagnostics but it encompasses all cases where the actual delivery of health care services involves a patient who is not located where the provider is. Its implementation requires the commissioning of an expensive telecommunications infrastructure and the exploration of a number of solutions. In fact, all categories of telemedical applications are considered while the full range of network choices is explored in terms of the cost/performance tradeoffs inherent to them and the developmental stage each of these options occupies in its life cycle. MERMAID utilises advanced land based line transmission technologies to aid the remote patient by making available the specialty care best suited to each particular case. Finally, the development of programme modules for training and education of the seafarers in the use of the MERMAID medical communications system is given top priority as such modules constitute the firmest basis for the promotion of proper practice of telemedicine at sea. PMID- 11010338 TI - Developing new competencies in healthcare practitioners in the field. AB - The Life-long Learning concept is one which is appropriate for those who have (or create) the opportunities to develop their competencies once they are in an operational role. All models of healthcare delivery and management are VOLATILE and therefore the way of addressing new competencies cannot be prescriptive or stand still for too long, but the concepts do endure. Learning, in order to 'keep up to date' is very necessary--technology, new clinical practices and interventions, new drug interactions, increased patient demands ... all add to the need to move with the times. This paper addresses some of the issues surrounding this challenge in the domain of informatics in support of healthcare delivery and management. PMID- 11010339 TI - Learning just-in-time in medical informatics. AB - Just-in-time learning (JITL) methodology has been applied to many areas of knowledge acquisition and dissemination. The paradigm is a challenge to the traditional classroom course-oriented approach with the aim to shorten the learning time, increasing the efficiency of the learning process, improve availability and save money. The information technology tools and platforms have been heavily involved to develop and deliver JITL. This paper discusses the main characteristics of JITL with regard to its implementation to teaching Medical Informatics. PMID- 11010340 TI - Education and training in health telematics. PMID- 11010341 TI - Prospects of medical informatics education in Greece: can we bridge the gap? AB - Medical Informatics education is a complex task,, since it aims in educating inhomogeneous groups of people coming from different disciplines. In Greece, recently considerable advancements in the medical informatics education field are made. These advancements aim in bridging gaps in various areas of MI users interactions. In particular, an overview of the strategy of the Greek ministry of education shall be given, some examples of MI related educational programs shall be referred to, and finally the MI educational structure used in the Aristotelian University shall be presented in detail, laying out the strategy of this University for bridging various types of gaps. PMID- 11010342 TI - Designing for interaction quality in health telematics. AB - This paper focuses on emerging Human Computer Interaction (HCI) challenges in the context of Health Telematics. Specifically, assumptions in traditional approaches to product and process quality are discussed, in order to unfold the reasons behind the loose and partial insight offered to the design of technology for the broadest possible end-user population. The premises of "Universal Design" are briefly discussed and the concept of "User Interfaces for All" is presented, as an articulation of Universal Design in the area of HCI. Following this, the paper discusses how such a perspective improves upon conventional methods and tools for HCI design and presents some of its relative merits in the context of Health Telematics. PMID- 11010343 TI - 21st century communication age. PMID- 11010344 TI - Use of multiple media and breast cancer screening: an introduction. PMID- 11010345 TI - Effect of media coverage and physician advice on utilization of breast cancer screening by women 40 years and older. AB - The purpose of this study is to examine the relative importance of media coverage and physician advice on the decision of women 40 years and older to obtain a mammogram. Five theoretical models, by which media coverage and physician advice may interact to affect individual health behavior, are presented. These models are tested with time-series regression analysis based on national-level data on mammography utilization and physician advice from the Behavioral Risk Factor Surveillance System (BRFSS) and content analysis of mammography-related national news coverage. Results suggest that although physician advice plays a key role in women's decisions to have mammograms, media coverage of mammography screening also contributes to mammography utilization by women. In particular, media coverage seems to be important for women who do not have regular contact with a physician or access to physicians. A possible conclusion is that mass media and physician advice complement one another in persuading individuals to adopt preventive health behavior. PMID- 11010346 TI - Experiences of women with breast cancer: exchanging social support over the CHESS computer network. AB - Using an existential-phenomenological approach, this paper describes how women with breast cancer experience the giving and receiving of social support in a computer-mediated context. Women viewed their experiences with the computer mediated support group as an additional and unique source of support in facing their illness. Anonymity within the support group fostered equalized participation and allowed women to communicate in ways that would have been more difficult in a face-to-face context. The asynchronous communication was a frustration to some participants, but some indicated that the format allowed for more thoughtful interaction. Motivations for seeking social support appeared to be a dynamic process, with a consistent progression from a position of receiving support to that of giving support. The primary benefits women received from participation in the group were communicating with other people who shared similar problems and helping others, which allowed them to change their focus from a preoccupation with their own sickness to thinking of others. Consistent with past research is the finding that women in this study expressed that social support is a multidimensional phenomenon and that their computer-mediated support group provided abundant emotional support, encouragement, and informational support. Excerpts from the phenomenological interviews are used to review and highlight key theoretical concepts from the research literatures on computer mediated communication, social support, and the psychosocial needs of women with breast cancer. PMID- 11010347 TI - Cigarette advertising to counter New Year's resolutions. AB - One process through which tobacco advertising may work is by reducing rates of quitting. Theories of addiction support the notion that relapse can be prompted by environmental cues. Further, because withdrawal symptoms occur over a predictable time frame, and because the most popular time to quit smoking is the beginning of the year, as a New Year's resolution, tobacco companies can make use of advertising to remind quitters of their need to smoke. Study 1 examined advertising in 10 popular magazines. It found a higher number of ads in January and February than the rest of the year after 1984. Study 2 examined cigarette advertising on the back cover of 10 other popular magazines. This study also found a higher rate of cigarette advertisements in January and February than for the rest of the year. The results suggest that cigarette marketers may be attempting to preempt quitting by cuing smoking behavior. PMID- 11010348 TI - Church-based telephone mammography counseling with peer counselors. AB - Little is published about step-by-step implementation of telephone counseling interventions to promote community-based health activities. This article describes the authors' experience of implementing a church-based telephone mammography counseling intervention with peer counselors representing three principal racial or ethnic groups: African American, Latino, and Anglo (White). Twenty-six women from 12 churches in the Los Angeles area were recruited and trained to deliver the counseling annually over a two-year period to 570 women participants who were recruited from participating churches (n = 15). The counseling sessions were conducted from church-based telephone centers in key geographic locations in our program area. Training and supervision proved challenging: most of the Latino counselors had fewer than seven years of education and spoke only Spanish, while most of the other counselors had at least some college and spoke only English. Culturally specific and small group interactions, role plays, and a more modular approach to training were the most effective ways to enhance counselors' skills. Latina participants' mammography adherence rates were lowest, and their barriers reflected their low socioeconomic status; as Latina counselors shared basic information about mammograms and where to obtain them at little or no cost, the counseling exchanges tended to be nonconflictive and supportive. Black and White participants were generally more knowledgeable and adherent with screening guidelines than Latinas. We found that it was possible to implement this intervention with diverse groups and conclude with lessons learned that may inform others considering such a strategy. PMID- 11010349 TI - Communications for HIV/AIDS prevention, care, and support: contexts for individual and social change. Introduction. PMID- 11010350 TI - Toward a new communications framework for HIV/AIDS. AB - In response to the overwhelming burden of new cases of human immunodeficiency virus (HIV) in Africa, Asia, Latin America, and the Caribbean, the Joint United Nations Programme on HIV/AIDS (UNAIDS), in 1997, initiated a project to examine the application of existing communication theories/models to HIV/acquired immune deficiency virus (AIDS) prevention and care in these regions. In the past 2 years, 103 leading researchers and practitioners from different parts of the world were invited by the UNAIDS to participate in one of five consultative workshops designed to review these theories/models and rethink their adequacy for Africa, Asia, Latin America, and the Caribbean. A new communications framework for HIV/AIDS was developed to move from a focus on the individual to a focus on five domains of "contexts" that influence behaviors: government policy, socioeconomic status (SES), culture, gender relations, and spirituality. PMID- 11010351 TI - From government policy to community-based communication strategies in Africa: lessons from Senegal and Uganda. PMID- 11010352 TI - Odo-Ya project: HIV/AIDS prevention in the context of Afro-Brazilian religion. PMID- 11010353 TI - Gender and HIV/AIDS in Africa--our hope lies in the future. PMID- 11010354 TI - Social and economic factors in an integrated behavioral and societal approach to communications in HIV/AIDS. AB - Health communication scholars have tried to understand how individuals process information and have identified the factors that contribute to appropriate behavior change. Some of these theorists have, implicitly or explicitly, assumed that if individuals were provided with the "right" information they would adopt the recommended behavior. Some others have endorsed the need to provide behavioral skills along with information so that individuals are able to carry out the desired behavior. Both approaches, however, are concerned with individual behavior change. Sociodemographic variables like class, gender, and race have seldom figured in sociopsychological analyses in the AIDS context. Limited attention has been paid to the manner in which political, economic, and social variables have constrained or enabled individual behavior related to AIDS. In this article, the various sociopsychological theories/models that inform AIDS prevention are delineated; the sociopsychological approaches in the context of class, race, and gender issues are critiqued; and an analytical framework that integrates behavioral and societal level variables to guide policy interventions is provided. PMID- 11010355 TI - HIV/AIDS communication campaigns: progress and prospects. AB - The mass media have been the primary method for disseminating human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) prevention messages worldwide. In this article, we update previous reviews by systematically examining published articles (n = 41) of empirical evaluations of international HIV/AIDS prevention campaigns. Of the 41 studies identified, 17 countries are represented. In this review, we examine six components related to media campaign design and evaluation: target audience, communication channel(s), message content, campaign theme, exposure, and outcomes. Of the studies included in this sample, each one described the target audience; 93% reported on channel selection; 75% described message content; 63% mentioned a campaign theme; and 62% documented campaign exposure. We investigate also the extent to which HIV/AIDS prevention efforts have moved beyond media campaigns to comprehensive communitywide programs. We conclude that HIV/AIDS prevention efforts would benefit from: (1) better reporting of media campaign components and outcomes, (2) more systematic evaluation, (3) greater integration of theory, and (4) increased attention to communitywide intervention strategies. PMID- 11010356 TI - Coping in cyberspace: the impact of Internet use on the ability of HIV-positive individuals to deal with their illness. AB - The human immunodeficiency virus (HIV) poses one of the greatest health threats of modern times. The Internet provides unparalleled access to resources that can assist individuals coping with HIV infection. Despite the promise of online resources to help individuals manage and cope with their illness, few studies have investigated the impact of their use. Those that have looked primarily at Internet use from a group perspective. These studies were usually "bounded" as well, focusing on electronic support groups or highly structured computer-based support systems that offer select users prescribed resource options. Little is known about how individuals who are "on their own"--without access to these services--use the vast, unstructured array of resources available through the Internet. The purpose of this study was to investigate the impact of Internet use on the coping ability of HIV-positive individuals. This research study employed a descriptive qualitative design that used indepth, semi structured, face-to-face interviews for data collection. The sample of 10 purposefully selected HIV positive individuals reflected diversity in gender, age, race, education, employment, number of years living with HIV, and Internet use. Data analysis guided by the constant comparative method revealed that the impact of Internet use on coping ability involved three themes: The Internet promotes empowerment, augments social support, and facilitates helping others. PMID- 11010357 TI - A critical assessment of theories/models used in health communication for HIV/AIDS. AB - Most theories and models used to develop human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) communication are based on social psychology that emphasizes individualism. Researchers including communication and health scholars are now questioning the presumed global relevance of these models and thus the need to develop innovative theories and models that take into account regional contexts. In this paper, we discuss the commonly used theories and models in HIV/AIDS communication. Furthermore, we argue that the flaws in the application of the commonly used "classical" models in health communication are because of contextual differences in locations where these models are applied. That is to say that these theories and models are being applied in contexts for which they were not designed. For example, the differences in health behaviors are often the function of culture. Therefore, culture should be viewed for its strength and not always as a barrier. The metaphorical coupling of "culture" and "barrier" needs to be exposed, deconstructed, and reconstructed so that new, positive, cultural linkages can be forged. The HIV/AIDS pandemic has served as a flashpoint to either highlight the importance or deny the relevance of theories and models while at the same time addressing the importance of culture in the development and implementation of communication programs. PMID- 11010358 TI - Peer promotion programs and social networks in Ghana: methods for monitoring and evaluating AIDS prevention and reproductive health programs among adolescents and young adults. AB - This article summarizes the observations and lessons learned regarding the application of human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) prevention and reproductive health evaluation methodologies in the context of adolescent and young adult populations and discusses the use of peer network evaluation to understand the dynamics of peer promotion. To examine the interpersonal communication process of peer education, this study tested a new approach using multiple semistructured interviews and network analysis to collect data on 106 peer educators and 526 of their contacts. These evaluation activities were conducted at three sites in Ghana during April 1998, in both periurban and rural locations in both in-school and out-of-school settings. By evaluating the social networks of peer educators it was possible to gain a better understanding of the process of peer education in terms of (a) defining the composition of peer contacts, (b) identifying the social norms that play a critical role in youth decision making, and (c) observing the range of messages and services transmitted during peer education. The objective of this paper is to disseminate the experience of the Center for Education and Development of Population Activities (CEDPA) and Focus on Young Adult's cooperative development of evaluation methodologies for peer promotion and to highlight utilization of these methodologies in a case study in Ghana. The results will be discussed in terms of their possible implications for program managers, researchers, and international agencies. PMID- 11010359 TI - Entertainment-education and HIV/AIDS prevention: a field experiment in Tanzania. AB - Entertainment-education is the process of designing and implementing an entertainment program to increase audience members' knowledge about a social issue, create more favorable attitudes, and change their overt behaviors regarding the social issue. The results of a field experiment in Tanzania to measure the effects of a long-running entertainment-education radio soap opera, Twende na Wakati (Let's Go with the Times), on knowledge, attitudes, and adoption of human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) prevention behaviors are presented. Multiple independent measures of effects and the experimental design of this study confer strong internal and external validity regarding the results of this investigation. The effects of the radio program in Tanzania include (1) a reduction in the number of sexual partners by both men and women, and (2) increased condom adoption. The radio soap opera influenced these behavioral variables through certain intervening variables, including (1) self-perception of risk of contracting HIV/AIDS, (2) self-efficacy with respect to preventing HIV/AIDS, (3) interpersonal communication about HIV/AIDS, and (4) identification with, and role modeling of, the primary characters in the radio soap opera. PMID- 11010360 TI - Protecting your paperless medical record system requires addressing risks. PMID- 11010361 TI - Terrorism: haz-mat bug for the new millennium. PMID- 11010362 TI - Developing a safe, effective restraint policy. PMID- 11010363 TI - Hot issues in clinical research. PMID- 11010364 TI - Evaluating your performance improvement system using a Baldrige-based self assessment. PMID- 11010365 TI - The importance of informed consent in medical research. PMID- 11010366 TI - Adoption of electronic medical records as a technology innovation for ambulatory care at the Medical University of South Carolina. AB - The study described in this article measured and compared the attitudes of groups of Medical University of South Carolina (MUSC) ambulatory care staff and physicians toward adopting an electronic medical record (EMR) system, using the Perceived Characteristics of Innovating (PCI) scales developed by Moore and Benbasat (1991). The PCI scale scores were compared by professional group and by clinic location. The overall findings of this study were that potential users of the ambulatory care EMR at MUSC had generally positive or neutral attitudes toward the system. There were a number of significant differences noted among the professional groups, particularly between the physician groups and other groups. The physician groups had less positive feelings than the other groups did. There were few significant differences noted for comparisons by clinic location. PMID- 11010367 TI - Cancer surveillance using data warehousing, data mining, and decision support systems. AB - This article discusses how data warehousing, data mining, and decision support systems can reduce the national cancer burden or the oral complications of cancer therapies, especially as related to oral and pharyngeal cancers. An information system is presented that will deliver the necessary information technology to clinical, administrative, and policy researchers and analysts in an effective and efficient manner. The system will deliver the technology and knowledge that users need to readily: (1) organize relevant claims data, (2) detect cancer patterns in general and special populations, (3) formulate models that explain the patterns, and (4) evaluate the efficacy of specified treatments and interventions with the formulations. Such a system can be developed through a proven adaptive design strategy, and the implemented system can be tested on State of Maryland Medicaid data (which includes women, minorities, and children). PMID- 11010368 TI - Organizational status of quality assurance and end-user perceptions of information technology for information management. AB - This article investigates the relationship between quality assurance (QA) and perceptions of the importance of information technology (IT) for information management in the health care industry. This issue is important because effective information management is recognized as a necessary condition for quality management. The unit of analysis of the article is the health care functional department. Data from 34 health care departments were collected and used for the analysis. Four specific hypotheses were proposed and tested. Our results suggest that end-user perceptions of the importance of IT for information management depends on formalization of QA, perceived importance of QA, time since QA was embraced, and number of people in the QA function. PMID- 11010369 TI - Big issues, small systems: managing with information in medical research. AB - This subject of this article is the design of a database system for handling files related to the work of the Molecular Genetics Department of the International Blood Group Reference Laboratory. It examines specialist information needs identified within this organization and it indicates how the design of the Rhesus Information Tracking System was able to meet current needs. Rapid Applications Development prototyping forms the basis of the investigation, linked to interview, questionnaire, and observation techniques in order to establish requirements for interoperability. In particular, the place of this specialist database within the much broader information strategy of the National Blood Service will be examined. This unique situation is analogous to management activities in broader environments and a number of generic issues are highlighted by the research. PMID- 11010370 TI - The disability assessment database model. AB - Collecting disability-related information in order to prevent and treat specific disabilities is important and necessary, but does it meet the major needs of individuals with disabilities? This article presents preliminary research performed to assess those needs as well as a Disability-service Assessment Database (DAD) model. This model provides local disability service data and surveys users so that their input is used to modify existing data. The content of the database and Web site are discussed and diagramed. A major goal of the DAD is to serve as a model to other communities. PMID- 11010371 TI - Health information management in the rehabilitation environment: the Easter Seal Society of western Pennsylvania. AB - Health information management functions are becoming more automated as facilities move toward the use of computerized health records. Nonprofit human services organizations are investigating the benefits of automating manual systems to determine the impact of automated systems on productivity, documentation, cost effectiveness, and quality of care. Management systems that integrate billing and clinical functions can be designed to meet the specific needs of any type of agency, such as rehabilitation agencies. This article discusses the initial implementation of such a system and its impact on staff and their work procedures. Also discussed is the agency's future vision of its computerized client management system. PMID- 11010372 TI - Managing the learning and work of new information technology users. AB - The information technology revolution has transformed work from modest-paced serial processes to complex, fast-paced networked ones that depend not only on the technology but also on skilled and effective users of the technology. This evolution has woven a complex quilt of interdependencies among new information technologies, new users of those technologies, new skills required, new information management policies and procedures, and new opportunities. We must impose a new structure for managing the learning and work of new users, their tolerance of complexity and continuous learning, value-added usability, minimization of human error, and new support services for deployment, administration, and help. PMID- 11010373 TI - Preparing for a decision support system. AB - The increasing pressure to reduce costs and improve outcomes is driving the health care industry to view information as a competitive advantage. Timely information is required to help reduce inefficiencies and improve patient care. Numerous disparate operational or transactional information systems with inconsistent and often conflicting data are no longer adequate to meet the information needs of integrated care delivery systems and networks in competitive managed care environments. This article reviews decision support system characteristics and describes a process to assess the preparedness of an organization to implement and use decision support systems to achieve a more effective, information-based decision process. Decision support tools included in this article range from reports to data mining. PMID- 11010374 TI - Justifying cardiac transports. PMID- 11010375 TI - Recent progress in advanced cardiac life support. AB - The revised guidelines for advanced cardiac life support (ACLS) from the American Heart Association are anticipated in the fall of 2000. Although dramatic changes in the approach to adult basic and ACLS are not anticipated, several controversies and new drugs on the horizon may radically change our approach to emergent cardiac resuscitation. This article features some of the evolving thinking on the emergent treatment of the adult with ventricular fibrillation or ventricular tachycardia, the critical rhythms seen in most cases of acute cardiac distress. Approaches to airway therapy drug administration and new agents also are described. PMID- 11010376 TI - Reperfusion of acute coronary syndromes and myocardial infarction. PMID- 11010377 TI - Chest pain: current concepts and implications for critical care transport. PMID- 11010378 TI - Oxygenation/ventilation of transported intubated adult patients: a national survey of organizational practices. AB - INTRODUCTION: Transporting intubated patients is common among ground and air crews, and providing adequate oxygenation/ventilation through transport ventilators (TV) or manual ventilation (MV) is clinically challenging. However, very little data chronicle service or practice patterns of oxygenation/ventilation within the industry. METHODS: During February 1998, a national sample of 250 air transport agencies was surveyed regarding activities and services surrounding this population of transported patients. One-hundred ninety-three surveys (77%) were returned. RESULTS: Approximately 40% of responding agencies use rotor-wing transportation only. Various combinations of rotor-, fixed-wing, and critical care ground transport were reported among the sample. Crew configuration consisted primarily of RN/EMT-P (75%). For pre hospital intubated patients, MV alone (37.3%), TV alone (10.9%), or a combination of MV and TV (43.5%) was used, depending on transport circumstances. Programs not involved in pre-hospital transports accounted for 8.3% of returned surveys. Interfacility transports used MV (6.8%), TV (39.4%), and a combination (53.4%). One respondent did not answer the question, accounting for 0.4% of the returned surveys. More than 75% of programs monitored oxygenation/ventilation during transport. Usually some combination of pulse oximetry and CO2 monitoring was used. More than half (59%) of reporting agencies transport more than 80 intubated adults each year. CONCLUSION: Considerable variation exists in practices involving the transport of intubated patients. PMID- 11010379 TI - Pilot study for predicting appropriate use of air medical helicopters. Part 1: Interfacility transports. PMID- 11010380 TI - New concepts in health benefits. Part 3. Employee incentives. PMID- 11010381 TI - Variety is the spice of a great workforce. PMID- 11010382 TI - There's no place like home ... for today's teleworkers. PMID- 11010383 TI - Are you paying too much for lab tests? PMID- 11010385 TI - DataWatch. Employers ask less about health care. PMID- 11010384 TI - Strength in service. PMID- 11010386 TI - Consumerism in the financial services industry: lessons for managed care. AB - Managed care today is being shaped by the emergence of a savvier, better informed health care consumer. Facing a strikingly similar consumer movement over the past two decades, the banking industry experienced a market transformation that holds important lessons for managed care. Nontraditional entrants in the financial services industry, offering focused "monoline" products and services closely analogous to "carve-out" providers in health care, targeted rising consumer demands and stronger preferences. Banks in time answered these formidable new competitive forces with innovative consolidation and globalization strategies. The most successful initiatives in healthcare, as in banking, will focus on satisfying the consumer's hunger for information, improved levels of service, and enhanced outcomes. Managed care plans may play a lead role in accelerating the impact of consumerism by bridging the disconnect between patients and their purchasing decisions. PMID- 11010387 TI - Customer value & business success in the 21st century. AB - Many health care organizations are realizing the importance of understanding the needs and expectations of customers and prospective customers, coupled with developing services that best meet and exceed those expectations. These concepts represent an innovative yet fundamental approach to building success beyond the clinical aspects. To create, build on, and measure the "customer value factor" in its bottom line, health care organizations must (1) understand how meeting and exceeding the needs and expectations of customers will become the most significant differentiator of true business value, (2) learn how to develop realistic, measurable, and successful customer value processes within complex organizations, (3) understand how customer value can link with the power of strategic communications to become the new currency for the next millennium. PMID- 11010388 TI - Customer service: the key to remaining competitive in managed care. AB - The health care industry is undergoing a rapid transformation to meet the ever increasing needs and demands of its patient population. Employers and managed care organizations are demanding better service and higher quality care, while providers are trying to tackle reimbursement cutbacks, streamlining of services, and serving a diverse population. Providers have begun to realize that to overcome these obstacles and meet the needs of their health plans and consumers, they must focus on the demands of their customers. Health care organizations have found they can meet the demands of both the consumer and the managed care industry through initiating and maintaining a customer service program. This essay explains the importance of customer service and its link to success in the managed care environment. PMID- 11010389 TI - The implementation of clinical guidelines in a managed care setting: implications for children with special health care needs. AB - OVERVIEW: The project tracked the development and implementation of a pediatric asthma guideline to determine its usefulness as a quality assurance mechanism for children with special health care needs. METHODS: Interviews were conducted with clinic staff to gather descriptive information about guideline implementation at owned clinics within a large HMO. RESULTS: Providers developed multiple implementation strategies emphasizing patient/family education. Service coordination within the health plan was well established, while coordination of services beyond the health plan was less clearly related to guideline implementation. CONCLUSION: Guideline implementation appeared to be a highly variable process. Clinical guidelines alone may not be sufficient tools of quality assurance for children with chronic or complex conditions. PMID- 11010390 TI - Health plan responses to Medicaid managed care policy in New York City. AB - Nearly all states in the United States have instituted managed care programs to serve Medicaid recipients and are developing policies to increase program participation. State practices regarding managed care contracting, premiums, and enrollment have implications for whether managed care plans will respond in a manner consistent with overall state policy objectives for the Medicaid managed care program. The experience of expanding the Medicaid managed care program in New York City, where the number of Medicaid beneficiaries exceeds the number in all but three states, has provided an interesting opportunity to look at the relationship between Medicaid managed care policy and plan enrollment. This paper analyzes trends in Medicaid managed care enrollment in New York City from January 1991 to September 1998, a period of critical changes in Medicaid managed care policy in New York. PMID- 11010391 TI - HMOs serving rural areas: experiences with HMO accreditation and HEDIS reporting. AB - This article explores the reasons why two groups of health maintenance organizations (HMOs) that serve rural areas have chosen to apply or not to apply for National Committee for Quality Assurance (NCQA) accreditation, and their future plans regarding accreditation. It also describes the challenges faced by HMOs serving rural areas in meeting NCQA accreditation standards and collecting Health Plan Employer Data and Information Set (HEDIS) data. A number of accredited HMOs cited positive benefits of the accreditation process, and most HMOs plan to apply or reapply for accreditation in the near future. Several HMOs identified challenges with accreditation standards, and the majority reported difficulty collecting some HEDIS measures. The HMOs identified a number of accreditation and HEDIS issues that are more problematic in rural areas. PMID- 11010392 TI - Ten top reasons teams become dysfunctional. PMID- 11010393 TI - The evolution of retirement: results of the 1999 Retirement Confidence Survey. AB - The ninth annual Retirement Confidence Survey (RCS) shows continued evidence of progress in the drive for retirement income security for American workers. However, there are still hurdles to overcome. The RCS tracks Americans' retirement planning and saving behavior and their confidence regarding various aspects of their retirement. It also categorizes workers and retirees into distinct groups based on their individual views on retirement, retirement planning, and saving. The retirement envisioned by today's workers looks different in many respects from that now experienced by current retirees. Today's workers expect to work longer than current retirees actually worked before retiring--and many say they plan to work for pay after they retire. Twenty-four percent of workers reported that they are very confident they will have enough money to live comfortably in retirement, and 45 percent reported that they are somewhat confident. However, there are indications that many may be falsely confident. The good news is that 70 percent of Americans are saving for retirement, and a growing percentage (49 percent) are going further and determining how much they need to save to fund their retirement. The bad news is that 30 percent of Americans have not begun to save for their retirement, and 51 percent have never tried to determine how much they need to save. Employers play a major role in ensuring adequate retirement preparation. Forty percent of all workers said they expect that money provided by their employer will be a major source of retirement income. Forty-six percent expect the money they put into a retirement plan at work to be a major source of income. The availability of a retirement plan at work is credited by 48 percent of savers as motivation to save. While worker education is a point of emphasis among both employers and policymakers, more remains to be done. For example, 59 percent of workers expect to be eligible for full Social Security benefits sooner than they actually will be, and an additional 19 percent admit they do not know when they will be eligible. There is evidence that education can have an impact on individual behavior. Forty percent of workers receiving educational material at work in the last year said that information caused them to begin saving (19 percent) or resume saving (21 percent) for retirement, while 40 percent said they changed the amount they were contributing to a retirement savings plan and 41 percent changed the allocation of their money in a retirement savings plan. PMID- 11010394 TI - Sources of health insurance and characteristics of the uninsured: analysis of the March 1999 Current Population Survey. AB - This Issue Brief provides summary data on the insured and uninsured populations in the nation and in each state. It discusses the characteristics most closely related to an individual's health insurance status. Based on EBRI estimates from the March 1999 Current Population Survey (CPS), it represents 1998 data--the most recent data available. In 1998, 194.7 million nonelderly Americans--81.6 percent- had some form of health insurance. More than 64 percent had it through an employment-based health plan; 6.5 percent purchased it on their own; and 14.3 percent were covered by a public program, mostly through Medicaid (10.4 percent). In 1998, 18.4 percent of the nonelderly population was uninsured (43.9 million people), compared with 14.8 percent in 1987. The percentage of uninsured Americans has generally been increasing since at least 1987, although the percentage uninsured in 1998 was not statistically different from the percentage uninsured in 1997 (18.3 percent). The increase in the uninsured prior to 1993 can be attributed to the erosion of employment-based health insurance. However, since 1993, the percentage of nonelderly Americans covered by an employment-based health plan has increased from 63.5 percent to 64.9 percent. The decline in public sources of health insurance would mostly explain the recent increase in the uninsured. For example, between 1994 and 1998 the percentage of nonelderly Americans covered by CHAMPUS/CHAMPVA declined from 3.8 percent to 2.9 percent, in large part due to downsizing in the military. Similarly, between 1993 and 1998, the percentage of nonelderly Americans covered by Medicaid declined from 12.7 percent to 10.4 percent as people left welfare. The increase in employment-based coverage since 1994 was due mainly to a higher likelihood that children were covered by an employment-based health plan. Between 1994 and 1998, the percentage of children covered by an employment-based health plan increased from 58.1 percent to 60.2 percent. For adults, it increased less than one percentage point, from 66.1 percent to 66.9 percent. Adults started to realize gains in employment based health insurance between 1997 and 1998. Between 1994 and 1997, the percentage of working adults with employment-based health insurance coverage held steady at roughly 72.3 percent. During this period, health care cost inflation was essentially nonexistent. However, between 1997 and 1998, the percentage of working adults with employment-based health insurance increased from 72.2 percent to 72.8 percent, despite the apparent return of health care cost inflation in 1998. It is likely that the changing composition of the labor force accounted for some of the increase in employment-based coverage. PMID- 11010395 TI - 401(k) plan asset allocation, account balances, and loan activity in 1998. AB - The Employee Benefit Research Institute (EBRI) and the Investment Company Institute (ICI) have been collaborating for the past three years to collect data on participants in 401(k) plans. This effort, known as the EBRI/ICI Participant Directed Retirement Plan Data Collection Project, has obtained data for 401(k) plan participants from certain of EBRI and ICI members serving as plan record keepers and administrators. The report includes 1998 information on 7.9 million active participants in 30,102 plans holding nearly $372 billion in assets. The data include demographic information, annual contributions, plan balances, asset allocation, and loans, and are broadly representative of the universe of 401(k) plans. The database also includes three years of longitudinal information on approximately 3.3 million participants. Key findings include: For all 401(k) participants in the 1998 EBRI/ICI database, almost three-quarters of plan balances are invested directly or indirectly in equity securities. Specifically, 49.8 percent of total plan balances are invested in equity funds, 17.7 percent in company stock, 11.4 percent in guaranteed investment contracts (GICs), 8.4 percent in balanced funds, 6.1 percent in bond funds, 4.7 percent in money funds, and 0.3 percent in other stable value funds. Participant asset allocation varies considerably with age. Younger participants tend to favor equity funds, while older participants are more disposed to invest in GICs and bond funds. On average, participants in their 20s have 62.1 percent of their account balances invested in equity funds, in contrast to 39.8 percent for those in their 60s. Participants in their 20s invest 4.7 percent of their assets in GICs, while those in their 60s invest 20.6 percent. Bond funds, which represent 4.7 percent of the assets of participants in their 20s, amount to 9.0 percent of the assets of participants in their 60s. Investment options offered by 401(k) plans appear to influence asset allocation. For example, the addition of company stock substantially reduces the allocation to equity funds and the addition of GICs lowers allocations to bond and money funds. The average account balance (net of plan loans) for all participants was $47,004 at year-end 1998, which is 26 percent higher than the average account balance at year-end 1996. The median account balance was $13,038 at year-end 1998. The balances, however, represent only amounts with current employers and do not include amounts remaining in the plans of prior employers. The average balances of older workers with long tenure indicate that a mature 401(k) plan program will produce substantial account balances. For example, individuals in their 60s with at least 30 years of tenure have average account balances in excess of $185,000. The ratio of account balance to 1998 salary varies with salary, increasing slightly as earnings rise from $20,001 to $80,000, and falling a bit for salaries greater than $80,000. The increase in ratio likely reflects a greater propensity of higher-income participants to save, whereas the decline after $80,000 results from contribution and nondiscrimination rule constraints. PMID- 11010396 TI - The working uninsured: who they are, how they have changed, and the consequences of being uninsured--with presidential candidate proposals outlined. AB - This Issue Brief presents data on workers who do not have health insurance. It offers a description of this population, discusses how this population has changed over time, and reviews the consequences of being uninsured. Also included is a description of the 2000 presidential candidates' proposals to reduce the number of uninsured Americans. The percentage of workers without any form of health insurance has been increasing since at least 1987. In 1998, 18.1 percent of workers were uninsured, up from 14.6 percent in 1987, although most of the increase occurred prior to 1993. Uninsured adult workers made up 56 percent of the uninsured population in 1998. In all, 83.2 percent of the 43.9 million uninsured Americans were in a family with a working family head. The working uninsured are heavily concentrated in certain segments of the population. In 1998, 53 percent of uninsured workers were under age 35, 58 percent were male, 57 percent were white, nearly 90 percent had not received a college diploma, 78 percent worked full time, 20 percent worked in the service industry, 60 percent were employed in small firms or were self-employed, 42 percent earned $7.00 or less per hour, and 99 percent earned less than $50,000 per year. The likelihood of being uninsured increased substantially for certain groups of workers between 1987 and 1998. The highest rates of increase were found among workers ages 55-64 (44 percent), in families at or above 400 percent of the federal poverty level (61 percent), in the public sector (34 percent), in the largest firms (53 percent), with hourly wages of $15.00 or more (50 percent), with annual income between $25,000 and $75,000 (100 percent), and with retirement plans (112 percent). Health insurance makes a difference in health status and access to health care services. Data show that uninsured workers are more likely than insured workers to report that their health status is fair or poor. Compared with insured workers, uninsured workers were more likely to receive health care in a hospital or emergency room, and were less likely to receive it in an office-based setting. Both the major-party presidential candidates, Vice President Al Gore and Texas Gov. George W. Bush, have put forth proposals to reduce the number of uninsured Americans. While both candidates' proposals recognize that the bulk of uninsured Americans are either children or workers employed by small firms, the proposed strategies to deal with these populations are incremental, and are unlikely to have a substantial impact on the number of uninsured Americans. PMID- 11010398 TI - Introduction to Employment 101: hiring the new grad. PMID- 11010397 TI - The maturing of consumerism. PMID- 11010399 TI - Customer service training ... the creative way! PMID- 11010400 TI - The next generation in health care. PMID- 11010401 TI - A commitment to satisfied customers. PMID- 11010402 TI - Look closely at responsibilities. PMID- 11010403 TI - House GOP unveils family safety priorities. PMID- 11010404 TI - Expanding the role of the board. PMID- 11010405 TI - Community benefits impact, Part 3. PMID- 11010406 TI - Putting patients and families first. PMID- 11010407 TI - Consumerism redefined ... the e-health imperative. PMID- 11010408 TI - Customer-centered redesign: a requisite for creating effective improvements. PMID- 11010409 TI - A stroll through the heart. PMID- 11010410 TI - Managing investment risk through asset allocation. PMID- 11010411 TI - Be an Internet millionaire and we may like you. PMID- 11010412 TI - High tech, high touch. PMID- 11010413 TI - Today's commitments required for tomorrow's success. PMID- 11010414 TI - Tomorrow is today. PMID- 11010416 TI - Champions of care. PMID- 11010415 TI - The 2000 Ludwig Award winners: setting the pace for a new millennium. PMID- 11010417 TI - Twenty-first century health care consumers. PMID- 11010418 TI - Rethinking rehab in the "post-BBA" market. PMID- 11010419 TI - Rethinking rehab: millennium market demands innovation, best practices, and consumer focus. PMID- 11010420 TI - Top 10 trends: rehab's "upside/downside potential" in the millennium. PMID- 11010421 TI - MedPAC begins work on SNF PPS updates; urges changes to quality monitoring, assessment tools. PMID- 11010422 TI - Preliminary Abt research points to four possible refinements to RUGS-III; HCFA proposal expected soon. PMID- 11010423 TI - Thinking of bringing therapy in house? Heed these considerations first. PMID- 11010424 TI - Underuse of beta-blockers in cardiovascular medicine. PMID- 11010425 TI - Cardiodynamic effects and mechanisms of action of beta-blockers. PMID- 11010427 TI - Dispelling the myth: incorporating beta-blockers into practice guidelines. PMID- 11010428 TI - How to achieve QI cooperation from health system physicians. PMID- 11010426 TI - The effect of beta-blockers on morbidity and mortality associated with heart failure. PMID- 11010429 TI - Big and small hospitals try benchmarking cooperative. PMID- 11010430 TI - Small facility explores benchmarking options. PMID- 11010432 TI - Inside Mecon's data mine: ground ambulance service. PMID- 11010431 TI - New system protects staff from pathogens. PMID- 11010433 TI - In tornado aftermath, Mitchell Country Hospital treats 200 people, scores 93 on JCAHO inspection. PMID- 11010434 TI - A case for rural hospitals: tornadoes may serve as wake-up call to state and federal lawmakers, rural citizens. PMID- 11010435 TI - Supreme Court says challenges to Medicare must initiate at HHS. PMID- 11010436 TI - A picture of teamwork: as tornados rip through four south Georgia counties, six hospitals rise to the occasion treating more than 120. PMID- 11010437 TI - Online tools help agencies cope with PPS. AB - A flurry of software development, much of it web-based, offers alternatives for agencies to address PPS requirements, streamline operations, and improve agency performance. PPS changes everything--dramatically altering the home care community's financial incentives and focusing attention on technologies that will enable agencies to succeed in this new environment. PMID- 11010438 TI - Running the numbers--is going high-tech in your agency's best interest? PMID- 11010439 TI - The Internet. Building knowledge & offering integrated solutions to health care. AB - Technology is changing the way we do things, including how the health care community provides information and services to the public. Disparate populations, once separated by distance and time, will experience dramatic changes as they become part of a global community. Fundamental changes will continue to occur in the way health information is received, utilized, exchanged and stored. This article explores access to the Internet as well as applications of interactive health technology and the emerging issues surrounding technology use. PMID- 11010440 TI - TeleHomeCare: connecting the home and the home care agency. PMID- 11010441 TI - Medical devices in the home: present and future applications. AB - Putting together a comprehensive telehealth program is similar to putting together a puzzle. You have to fit all of the pieces together to see the big picture. An important piece of that puzzle is medical devices for telemonitoring. The equipment used in a telehealth program can span from low end, such as simply using a telephone; to high end, in which two-way audio-video systems are used with computer documentation, outcome software, and various medical device options. PMID- 11010442 TI - The new hospice compliance plan: defining and addressing risk areas. Part 3. AB - The recently released OIG guidelines to ensure compliance with federal and state statutes, rules, and regulations, and private-payor health care program requirements provide a blueprint for developing such programs. This is the last of three installments that focus specifically on the 28 risk areas identified in the guidance and offer strategies for incorporating them in a hospice compliance program. The authors have organized the 28 risk areas under 9 topic domains to simplify the task of tackling the guidance. This article covers the areas of nursing home care, marketing, and Conditions of Participation. PMID- 11010443 TI - Partnering to prepare for success. PMID- 11010444 TI - Exploring the new frontier: home care gets wired. AB - The driving forces toward adoption of TeleCare into the non-acute care setting are more compelling than ever before. The dramatic increase in our elderly population, which provides an endless consumer base, availability of functional and affordable technologies, and a national mandate to control costs of health care are forcing even the most skeptical health care decisionmakers to accept TeleCare as a standard model of care. Add to this the most severe nursing shortage to date and the formula is complete. PMID- 11010445 TI - Managed care liability today: laws, cases, theories, and current issues. AB - Managed care entities face numerous liability issues in today's changing healthcare environment. This Article provides the plaintiff with a comprehensive road map for navigating the many avenues of managed care liability. The author describes ERISA pre-emption provisions and suggests ways plaintiffs' attorneys can strive to narrow the pre-emption. The Article also provides in-depth analysis of each theory of managed care liability that has been litigated against managed care entities to date, and then goes on to explore state laws imposing liability on managed care entities, and how HMO liability is being reformed through legislative action. For plaintiffs' attorneys seeking the full spectrum of theories of managed care liability, or for defendants' attorneys wanting to remain updated on all potential claims to defend, this Article constitutes an extensive primer on the current issues. PMID- 11010447 TI - Consumer managed care appeals: are the available procedural protections fundamentally fair? AB - Managed care incentives to reduce costs have also resulted in incentives to deny care. Anecdotes concerning managed care denials of care have led to a consumer outcry for protection either through the use of procedural due process or by the establishment of patient rights that would include appeal and grievance protections. This Article reviews the procedural protections of constitutional due process, the Consumer Due Process Protocol, and the Patient Bill of Rights. The Article then extensively discusses the availability of these procedural protections in various public and private forums. The discussion of public forums includes proposals contained in recent national legislative initiatives. The author then reviews relevant federal and state law, as well as Uniform Law proposals. Next, the Article analyzes the protections provided by accreditation agencies, dispute resolution organizations, professional organizations, and health insurers. Finally, the author recommends criteria to be used to determine whether a procedure is fundamentally fair. PMID- 11010448 TI - The National Practitioner Data Bank: coping with the uncertainties. AB - The National Practitioner Data Bank is a database of adverse events involving physicians and other practitioners. Querying the database is mandatory for hospitals in several situations. So too, hospitals are required to report specified adverse events. Thus, hospitals need to be able to identify incidents that are reportable events, events that require them to update the databases, and any possible liability issues that may surround the hospital's reporting duties. The author argues that the regulations are unclear in addressing these issues. Likewise, he notes that practitioners should be aware of other problems with the reporting system, including the lack of sufficient Data Bank security. PMID- 11010446 TI - Creating problems as part of the "solution": the JCAHO Sentinel Event Policy, legal issues, and patient safety. AB - The recently reported number of patient deaths attributable to medical errors is staggering. In response to this crisis, the Joint Commission on Accreditation of Healthcare Organizations ("JCAHO") proposed its Sentinel Event Policy. The policy mandates self-reporting by hospitals accredited by the JCAHO. This Article argues that the JCAHO's policy shows an inattention to the legal realities of discovery and absence of immunity. Until the JCAHO addresses these issues, self-reporting will have limited success. This Article suggests that to promote patient safety, self-regulatory reports should go to a neutral, nonsanctioning third party, an approach adopted from aviation's highly successful reporting system. PMID- 11010449 TI - Managing the outpatient data grouping process. PMID- 11010450 TI - Freestanding continuing care facility offers lessons in how to keep customers happy. PMID- 11010451 TI - McManis: technology, consumerism keys to a successful future U.S. health system. PMID- 11010452 TI - Flurry of solicitous e-health businesses offers hospitals plenty of alternatives. PMID- 11010453 TI - BBA pain gets worse, new studies show; pinch not likely to go away, EY consultant says. PMID- 11010455 TI - Telehealth expands unrealized dream. PMID- 11010454 TI - Failure of AHERF offers plenty of lessons in how not to run an integrated health system. PMID- 11010456 TI - A visit to a Mexican specialty hospital: the patient as leader? PMID- 11010457 TI - Looking for Dr. Jekyll but hiring Mr. Hyde: preventing negligent hiring, supervision, retention, and training. PMID- 11010458 TI - Misconceptions regarding restructuring of registered nurse positions in western New York and southern Ontario. PMID- 11010459 TI - Acquired immunodeficiency syndrome (AIDS) update, Part 2. PMID- 11010460 TI - Economic development: a moral imperative for creating flourishing communities. PMID- 11010462 TI - Price survey. Market forces squeeze stent prices. PMID- 11010461 TI - Preventing needlestick injuries in healthcare settings. PMID- 11010463 TI - E-commerce poses more questions than answers. PMID- 11010464 TI - Transforming health services management education and development: a challenge for the new millennium. PMID- 11010465 TI - Evolving health services administration education: keeping pace with change. AB - Virginia Commonwealth University (VCU) celebrated its golden anniversary in 1999. It was an occasion for faculty and alumni to renew memories of their experiences together and marvel at the dramatic changes they have seen in the health care industry. This qualitative study was undertaken to reflect on the department's immediate future course, and to learn how to reposition the education program and curricula to respond to evolving industry needs and demands. Results and conclusions are based on interviews conducted with several stake-holder groups, including VCU alumni, who are executives in the health sector; human resources recruitment officers; and selected program directors from leading health administration (HA) education programs in the U.S. Trends are identified in the health care industry environment, and strategies are proposed for the HA education programs to strategically adapt for the future. PMID- 11010466 TI - Perceived effectiveness of teaching methods for heath services administration education. AB - This article brings to the forefront the challenges of establishing an empirical link between teaching methods and education outcomes in the classroom. In the health administration education field, precious little empirical data exist on the effectiveness of various teaching methods and approaches on achieving education outcomes. Results are reported from the 1998 National Survey of program directors of the AUPHA graduate programs on the perceived effectiveness of a wide range of teaching methods, using a Likert scale, ranging from very effective to not very effective. The authors elaborate on the highest- and lowest-ranked teaching methods, and propose a research agenda for measuring education outcomes in the classroom. PMID- 11010467 TI - The role of accreditation in transforming health services management education. AB - The core function of the Accrediting Commission on Education for Health Services Administration (ACEHSA) is to improve quality in health services management education. All specialized accrediting agencies provide quality assurance by developing standards of quality, assessing programs and reporting to stakeholders, as part of their public accountability function (Council for Higher Education Accreditation). Standards of quality derive from the level of competency considered essential within the domain of practice of health services managers. Accreditation review by ACEHSA is oriented to quality improvement and quality assurance, which have different purposes and are difficult to accomplish in the same process. The purpose of this paper is to discuss ACEHSA's role in evaluating the quality of health services management education programs in North America. The first objective is to describe the process of defining quality and to identify the skills, knowledge and abilities derived from it. The second objective is to discuss the measurement of quality, including using standardized measures, and determining whether the purpose is quality improvement or quality assurance. The third objective is to examine how quality measures are reported, and how reporting impacts the quality assessment process. Finally, the future role of ACEHSA is discussed, with regard to benchmarking measures of quality and publicly reporting relative levels of academic program quality. PMID- 11010468 TI - A framework for evaluating the impact of health services management education. AB - The objectives of this paper are three-fold. The first is to propose a conceptual and analytical framework for planning and evaluating health services management education programs. The framework was developed in response to the health care industry restructuring and the emerging questions and concerns regarding how to strategically redesign the educational programs for the twenty-first century. Additionally, it was used to develop a questionnaire for the 1998 National Survey of Program Directors. The second objective is to suggest variables for operationalizing the framework as the field considers initiatives such as, "Evidence-Based Health Administration Education" (AUPHA Education Outcomes Task Force 1999; Griffith 2000) and other more standardized, quantifiable approaches to assessing program quality. The third objective is to report selected results from the 1998 National Survey of Program Directors focusing on the major changes impacting the health services industry and the implications for management education and development. In the future, ACEHSA and other accrediting commissions will likely consider evidence-based evaluation criteria for assessing and improving education program quality. The framework presented in this paper can be used as a starting point for conceptualizing and developing valid and reliable measures. PMID- 11010469 TI - How location can make or break your practice. PMID- 11010470 TI - I opened my home to a homeless patient. PMID- 11010471 TI - Keep up with old friends--but don't treat them. PMID- 11010472 TI - The new rules on informed consent. PMID- 11010473 TI - Legislators begin their effort to curb medical errors. PMID- 11010474 TI - The managed care program isn't working the way Congress intended. PMID- 11010475 TI - Risk adjustment: Medicare's latest move to tinker with your income. PMID- 11010476 TI - Make your office layout serve your needs. PMID- 11010477 TI - The snake in the ER. PMID- 11010478 TI - Attacking managed care with a full-court press. PMID- 11010479 TI - Where students pledge themselves to primary care. PMID- 11010480 TI - "Don't let go". PMID- 11010481 TI - Thank you, Carlos, for rekindling my love of medicine. PMID- 11010482 TI - Disabled by my illness and my insurer. PMID- 11010483 TI - How to squash a small malpractice claim. PMID- 11010484 TI - Why didn't I speak up for this patient? PMID- 11010485 TI - A managed care sting. PMID- 11010486 TI - Helping the homeless move ahead with their lives. PMID- 11010487 TI - Offering medical advice on the Web. PMID- 11010488 TI - Will the Internet finally put an end to paperwork? PMID- 11010489 TI - How connectivity is changing practice. PMID- 11010490 TI - Professional liability. 1998: a year when malpractice awards mushroomed. PMID- 11010491 TI - They're still working on the electronic medical record. PMID- 11010492 TI - Medicare's new red flag: same-day readmissions. PMID- 11010493 TI - When to hire another doctor--and how to make it pay. PMID- 11010494 TI - Find a doc, keep a doc, keep a doc happy. PMID- 11010495 TI - How unhappy are women doctors? PMID- 11010496 TI - How I almost killed Otis. PMID- 11010497 TI - The biggest coding blunders--and how to avoid them. PMID- 11010498 TI - Congress backs away from mandatory reporting of medical errors. PMID- 11010499 TI - Managed care reform? Sure--but don't look to Washington. PMID- 11010500 TI - How to harmonize with PAs and NPs. PMID- 11010501 TI - Things to do in Denver when your practice is dead. PMID- 11010502 TI - Get the most out of every patient visit. PMID- 11010503 TI - Quality facility standards: more than the right thing to do. PMID- 11010504 TI - Developing a Web site for your practice. PMID- 11010505 TI - Dynamic modeling of outpatient systems to balance provider resources and patient access. AB - Improving patients' access to appointments remains a major goal for most managed care organizations and group practices. A number of factors affect the length of time a patient must wait to see a physician or other provider. This article describes those factors and how dynamic modeling can assist decision makers in determining the appropriate number of providers or changes in practice or operations needed to meet given access targets. PMID- 11010506 TI - Fixing responsibility for risk management. AB - The responsibility for carrying financial risk for medical coverage has migrated from individuals to insurers to employers to providers, without finding a satisfactory home. Each shift further complicates the health care infrastructure, as other responsibilities in the management of benefits and provision of care gravitate to the stakeholder who accepts risk. The social imperative to broaden coverage is forcing a change in the mechanisms of risk management--from avoiding high-risk patients, to managing those patients to better outcomes. In this paper we seek to identify objectively the most appropriate party to carry the financial risk of medical coverage, consider what characteristics are necessary to make that a practical and enduring solution, and examine the secondary effects of the structure required to support that solution. PMID- 11010507 TI - An academic practice's transition to the business of medicine in the community. A case study. AB - This case study highlights the problems confronting a clinical practice corporation affiliated with a major medical school, and the business realizations it made in the acquisition of a community-based clinic. Launching a financially viable enterprise requires careful planning, determination of formal goals and expectations, an appropriate mix of physicians and services, a specific marketing campaign and community support. PMID- 11010508 TI - When you suspect the healer needs healing. AB - By recognizing the causes of stress among physicians in your office or hospital and proactively addressing them, you can play a significant role in improving patient care. PMID- 11010509 TI - Ready: how to keep your customers coming back. AB - Customer service is a major, but often overlooked, issue in health care today. While other industries and organizations recognize how good customer relations can affect long-term success, many health care providers have yet to learn this valuable lesson. The Ritz-Carlton, which won the prestigious Baldridge Award for service, has a well-earned reputation for excellent customer service. Like health care providers, this hotel industry icon hires hourly workers, puts them in uniform and has them work in teams. Unlike health care, however, The Ritz-Carlton seems to be able to generate a much higher level of customer satisfaction. How? This chapter illustrates the techniques the hotel chain uses to accomplish its goal and how these important tools can apply to the health care industry. PMID- 11010510 TI - Kaiser on track to recovery. PMID- 11010511 TI - Top court toughens rules. Providers can't take Medicare grievances directly to federal court. PMID- 11010512 TI - St. Vincent's is one dumped in breakup. PMID- 11010514 TI - Late, but online. Big group purchasing organizations play catchup in bid to place services on Internet. PMID- 11010513 TI - AHA backs bill to end key disparities. PMID- 11010515 TI - More BBA relief not likely. PMID- 11010516 TI - Settlements may not unbumdle. Appeal decision notwithstanding, hospitals unlikely to get fully repaid in fraud probe. PMID- 11010517 TI - Doc contracts under fire in Calif. PMID- 11010518 TI - Worst-case scenario. Atlanta hospital's demise, despite powerful backing, makes it poster child for industry woes. . PMID- 11010519 TI - Ga. hospital barely stays afloat. PMID- 11010520 TI - Taking a stand is easier in a crowd. PMID- 11010521 TI - What hospitals 'see' they get. Private sector acquiesces to providers' price hikes. AB - With government reimbursement continuing to be the target for budgetary squeezing, hospitals are turning to the only place where they can raise prices: the private sector. There's mounting evidence that hospitals are exacting higher rates from health plans and other nongovernmental payers, marking a resurgence of cost-shifting following a five-year hiatus. PMID- 11010522 TI - Providers put faith in Internet. PMID- 11010523 TI - Care on the frontier. Alaska hospital meets challenges not found in 'lower 48'. PMID- 11010524 TI - Patients get choices in drug pricing. PMID- 11010525 TI - Aching backs a blooming business. PMID- 11010526 TI - Credit crunch. Many healthcare companies are discovering a good lender is hard to find. PMID- 11010527 TI - Premier's feeling lucky. PMID- 11010528 TI - La. providers challenge big Medicaid cuts. PMID- 11010529 TI - 'Statistically insignificant' but critical. Increase in improper Medicare payments weakens healthcare providers' case for BBA relief. PMID- 11010530 TI - Ohio system plans expansion. Mount Carmel challenges OhioHealth in quest to be state's largest system. PMID- 11010531 TI - O'Leary clarifies the JCAHO's stance. PMID- 11010532 TI - New leaders drive managed care. Sensitivity with a bulletproof attitude required. AB - It's been a time of rapid-fire change in the managed-care industry--change that has included high-profile turnover in the executive suites at many of the large publicly held HMOs. Stepping into the top jobs has been a new generation of leaders with skills and qualities unheard of in boardrooms just a decade ago. But those skills will be tested as the industry continues to evolve. PMID- 11010533 TI - Reports find error-reporting takes faith. PMID- 11010534 TI - 2000 construction & design survey. PMID- 11010535 TI - AHERF executives arrested. Pa. attorney general says three stole endowment funds. PMID- 11010536 TI - Aetna will go it alone. PMID- 11010537 TI - Heart palpitations. Development of cardiac specialty-care hospitals often leads to bitter market feuds. PMID- 11010538 TI - Growth but no margins. PMID- 11010539 TI - Joint venture creates cancer center. PMID- 11010540 TI - Healthcare agenda puzzles Congress. PMID- 11010541 TI - Healthcare becoming a lot less local. PMID- 11010542 TI - Heavyweight hospitals build strength from within. Columbia, Tenet concentrate on core groups of hospitals. PMID- 11010543 TI - Do-it-yourself purchasing. PMID- 11010544 TI - Bankruptcies' ripple effects. PMID- 11010545 TI - A new revenue source. PMID- 11010546 TI - MedPAC blames private payers. . PMID- 11010548 TI - AMA cancels accreditation program. PMID- 11010547 TI - Vencor walloped for $1.3 billion. PMID- 11010549 TI - Double indemnity. PMID- 11010550 TI - Credentialing in a vacuum. PMID- 11010551 TI - A 'bad news sells' strategy. PMID- 11010552 TI - Betting for a no-show. PMID- 11010553 TI - Drug-free in Seattle. Harborview Hospital takes on heroin. PMID- 11010554 TI - Another bailout looming? PMID- 11010555 TI - Fiscally challenged. Psychiatric industry's hope is in consolidation, focusing services. AB - Psychiatric hospitals are closing in droves. Last year alone, at least 75 were shuttered, up sharply from the previous year. And psych hospitals collectively lost $254 million in 1998. To reverse the sector's fortunes, providers are focusing on consolidation and specialty services. "Mental health isn't glamorous," says David Moulthrop (left), CEO of a Wisconsin psych hospital. "But there is a future, though the pie has shrunk." PMID- 11010557 TI - Still governing. PMID- 11010556 TI - A leader who mixes quality, profitability. PMID- 11010558 TI - Taking a hike. PMID- 11010559 TI - Small businesses fight Fla. insurers. PMID- 11010560 TI - Use of locum tenens grows. PMID- 11010561 TI - Challenging the REITs. Real Estate Investment Trusts. PMID- 11010562 TI - Little change likely after rating firms' merger. PMID- 11010563 TI - The politics of health. PMID- 11010564 TI - A comparison of the awareness and utilisation of postoperative health services provided to women with breast cancer in public and private hospitals. AB - Nurses at a metropolitan Cancer Care Centre (CCC) noted that women who have had recent breast surgery for carcinoma had significantly different levels of knowledge and use of support services depending upon public or private hospitalisation. The public hospital participants were more aware of the range of available services (mean = 3.6) compared to women from the private sector (mean = 2.6). In addition, the public hospital participants were more likely to access a wider range of services post discharge (mean = 2.21) compared to the private hospital women (mean = 0.85). A significant difference was found between younger and older women's use of services. PMID- 11010565 TI - Improving local services for women with breast cancer: interviews with general practitioners in central Sydney. AB - We conducted interviews with 85 randomly selected general practitioners (GPs) in Central Sydney to examine patterns of referral of women with breast cancer, satisfaction with local services, awareness of evidence-based guidelines and suggestions for local support. Previous experience was the most frequently cited factor influencing choice of specialist (n = 80, 94%) followed by personal knowledge of the consultant's expertise (n = 72, 85%). Only one third of respondents agreed women wanted to be more involved in choosing the specialist (n = 28, 33%). Of 79 women recently diagnosed with breast cancer, the majority (96%) had been referred to a male surgeon (n = 71, 96%). While only 35% of the GPs rated the care received by women with breast cancer in local facilities as 'excellent', significantly fewer rated communication between GPs and specialists as 'excellent' (35% v 19%, p < 0.01). Younger GPs were less likely to rate communication as 'excellent' compared with GPs aged 55 years or older (p = 0.01). Only 18% of GPs considered their patients as having been 'very well informed' about their treatment choices. Younger GPs were more likely than older GPs to recall all available breast cancer guidelines (p = 0.02). Significantly more GPs (68%) requested seminars with experts than any other types of educational support (p < 0.001). To improve outcomes for women with breast cancer, mechanisms to support communication between GPs and specialists are recommended. Seminars for GPs with experts who emphasize evidence-based guidelines should be funded and evaluated, especially for impact in meeting the needs of older GPs. PMID- 11010566 TI - An assessment of the needs of oncology outpatients for the development of allied health services. AB - This article presents the findings of a study that arose out of the desire by the Allied Health Professionals (AHP) at the Mater Hospital, Brisbane, to better understand the needs of their clients in order to be able to offer a more effective and appropriate service. A questionnaire designed specifically to explore the needs of patients and their families for AHP services was administered to consecutive patients (n = 62) attending the Mater out-patient oncology clinic during one month. The findings provide a wealth of practical information for AHPs to use in planning for the effective utilisation of their services, as well as fresh insights into a number of theoretical issues that need further research. PMID- 11010567 TI - Emergency medicine and "acute" general practice: comparing apples with oranges. AB - Emergency Departments (EDs) operate at the interface between the inpatient and ambulatory sectors of health care. Because of shared funding between the Commonwealth and States for ambulatory care, there has been intense focus on the ED patient population, and the potential to shift the locus of care for non inpatients. One of the frequently cited models for the provision of after-hours GP services is the Balmain General Practice Casualty (GPC). This paper analyses the GPC model, looking in detail at casemix, clinical quality, waiting times and cost-effectiveness. It is argued that the services provided and the casemix of the patient population of GPC and EDs are distinctly different. Cost effectiveness for GPC has not been objectively established. Health service planning should recognise the distinct but complementary roles of general practice and emergency medicine. Evaluation of alternative models of service provision should critically examine the available evidence, and comparisons should be based on a precise analysis of equivalent services. PMID- 11010568 TI - Home phototherapy for neonatal jaundice--technology and teamwork meeting consumer and service need. AB - 32 babies with uncomplicated physiological jaundice received home phototherapy from a hospital/community team in southern Brisbane. All babies showed acceptable reductions in their serum bilirubin on home therapy, and none required hospital re-admission for phototherapy. Their families were highly satisfied with the home program, and recorded high levels of confidence in their therapeutic responsibilities. Community providers were able to deliver a high quality 24-hour service, integrated with appropriate neonatology support. The cost of delivering the home program was significantly less than a comparable hospital stay. PMID- 11010569 TI - Improving clinical indicators in acute admissions to the Department of Geriatric Medicine, Royal Perth Hospital. AB - Clinical indicators are an important component of quality assessment of clinical services. We outline the strategies used in the department of Geriatric Medicine at Royal Perth Hospital (RPH) to report on and improve the results. The clinical indicator for assessment of cognitive function had improved from 19% in September 1998 to 64% in February 1999. The clinical indicator for assessment of physical function has been maintained at 80%. There have been revisions to the definitions of the clinical indicators for 1999. The current clinical indicators used in this department can be modified for comparison nationwide amongst geriatric units. PMID- 11010570 TI - Developing mental health rehabilitation services in a culturally appropriate context: an action research project involving Arabic-speaking clients. AB - The present study illustrates the efforts by a community mental health service to restructure the Mental Health Rehabilitation Program to meet more adequately the needs of the Arabic-speaking population. It was discovered that the principles of mental health rehabilitation theory are inherently Anglo-Australian and not suitable for the Arabic-speaking population. Action research methodology was used as a tool to drive the changes necessary to implement effective mental health rehabilitation across cultures. Two new culturally relevant programs were developed, staff attitudes to rehabilitation were changed and assertive strategies were introduced to alter clients' attitudes and perceptions about mental health rehabilitation. PMID- 11010571 TI - Nurse-managed analgesia for renal colic pain in the emergency department. AB - A retrospective chart review was conducted of patients with acute renal colic for the years 1993 and 1997, in order to compare analgesia ordering and administration practices before and after implementation of a nurse-managed, titrated intravenous (i.v.) narcotic policy. The study demonstrated a significant and sustained change in analgesia administration practices away from the intramuscular (i.m.) route in favour of the i.v. route. For renal colic, in 1993, 76% of patients received i.m. narcotic analgesia compared to 3% in 1997. In contrast, i.v. narcotic (with or without adjuvant (NSAID) was used in 3% of the patients in 1993 compared to 95% in 1997. PMID- 11010572 TI - Improving the utilisation of bilingual counsellors within a public sector mental health service. AB - An urban, public sector Area Mental Health Program reviewed its own bilingual counsellor program as part of an Area-wide quality improvement project, and found that the counsellors' roles needed to be better defined; that mainstream staff needed to have more access to their expertise as cultural consultants; and that their function as an Area team, rather than as service based staff, needed to be encouraged. The bilingual counsellors decided to take up this challenge, and with the support of the Area Director of Mental Health, worked together to redefine their roles. PMID- 11010573 TI - Managed competition. PMID- 11010574 TI - Independent life expectancy in New Zealand, 1996-97. PMID- 11010575 TI - The performance of Australian DRGs. AB - This paper reports the results of an evaluation study of the Australian National Diagnosis Related Groups (AN-DRGs). The evaluation was based on statistical rather than clinical criteria with the principal goal being to provide information for the future development of the classification system. As well as comparing versions 1.0 to 3.0 of AN-DRGs, the project included a comparison of these systems with the most recent versions of the DRG systems from the United States. Taking all the evaluation criteria together, Version 3.0 of AN-DRGs performed best of all the systems except for the All Patient Refined (APR)-DRGs with its much larger number of groups. However, the differences between all the classifications were slight. Data of higher quality are needed if further refinements of the AN-DRGs are to produce substantial improvements in performance. PMID- 11010576 TI - EquIP accreditation: feedback from a Sydney teaching hospital. PMID- 11010577 TI - Taking health seriously: the senate inquiry into public hospitals. PMID- 11010578 TI - Distributional impact of higher patient contributions to Australia's Pharmaceutical Benefits Scheme. AB - This paper uses NATSEM's Pharmaceutical Benefits Model to analyse the effects of a hypothetical 25 per cent rise in patient contributions to prescribed medicines under Australia's Pharmaceutical Benefits Scheme (PBS). The model, based on microsimulation techniques, is able to provide a much broader range of outcomes information, at a much greater level of detail, than is possible with traditional methods. Higher patient contributions are analysed in terms of their impact on the government to patient split in PBS costs, as well as the distribution of such costs across age groups, family incomes, family types and 36 prescribed medicine types. Also considered are changes in the shares of family disposable incomes spent on prescribed drugs arising from the higher patient contributions. PMID- 11010579 TI - High length-of-stay outliers under casemix funding of a remote rural community with a high proportion of aboriginal patients. AB - The diagnosis related groups (DRG) classification was designed primarily to categorize patients of acute short-stay hospitals in urban areas. As one might expect, many studies have shown it is a less effective predictor of the needs- and consequently the costs of care--of remote and socio-economically disadvantaged communities. One way of improving the equity of funding involves separating the cases in each DRG into inlier and outlier episodes, and making different resource allocations for each category. This paper summarises the outlier payment model used by the Health Department of Western Australia, with emphasis on high length of stay outliers. The model provides additional funds for high length of stay outliers, but funding levels are deliberately set below the actual estimated costs of care, on the assumption that some of the additional costs are a consequence of poor care management. All high length of stay outlier episodes in the East Pilbara Health Service in 1997-98 were examined. It was found that the outliers were predominantly Aboriginal patients from remote communities with higher than average needs for care as indicated by their greater tendency to have multiple conditions requiring treatment. The age distribution of high length of stay outliers was quite different from that found in most Australian hospitals, in that there was a higher proportion of young children. It is concluded that, although the ideas on which the funding model is based are sound, revisions of detail need to be considered to reduce the risk that the burden of cost containment will fall to a disproportionate degree on the most disadvantaged groups of patients. PMID- 11010580 TI - Implications of decreasing surgical lengths of stay. AB - A recent study at the Prince of Wales Hospital (PoW) compared health outcomes and user satisfaction for conventional clinical pathways with a shortened pathway incorporating day of surgery admission (DOSA), early discharge and post acute care domiciliary visits for two high volume, elective surgical procedures (herniorrhaphy and laparoscopic cholecystectomy). This paper quantifies cost differences between the control and intervention groups for nursing salaries and wages, other ward costs, pathology and imaging. The study verified and measured the lower resource use that accompanies a significant reduction in length of stay (LOS). Costs of pre- and post-operative domiciliary visits were calculated and offset against savings generated by the re-engineered clinical pathway. Average costs per separation were at least $239 (herniorrhaphy) and $265 (laparoscopic cholecystectomy) lower for those on the DOSA pathway with domiciliary post acute care. PMID- 11010581 TI - Clinical pathways: a direction forward in health care. AB - As in many developed nations, health care consumers in Singapore are demanding better (and more costly) services. At the operational level, one way to address consumers' demands while ensuring cost-effectiveness is through the implementation of a clinical pathway program. This paper provides an overview of the program at Changi General Hospital (CGH), a regional general acute care hospital in the Republic of Singapore. The paper highlights the problems encountered during the planning, developing and implementing phases. It concludes by predicting what the future might hold for clinical pathways in Singapore and the South East Asian Region. PMID- 11010582 TI - Clinical pathways involving general practice--a new approach to integrated health care? AB - The Mater Mothers' Hospital, South Brisbane recently identified a number of difficulties with the maternity share-care program it runs with 1100 local GPs. This paper describes an integration approach developed at the Mater which has addressed these problems via the use of clinical management guidelines across the whole episode of care, the provision of a patient held record/pathway as a clinical practice prompt, clear communication and information management protocols between hospital and general practice, and the provision of continuing medical education for share-care practitioners. PMID- 11010583 TI - WESTCOP: a disease management approach to coronary artery disease. AB - Disease management is a systematic approach to improving care of populations of patients with specific clinical conditions. Critical to success are the formation of collaborative teams of health care stakeholders, development and promulgation of clinical practice guidelines, and performance measurement and feedback to providers as a process of continuous practice improvement. This article describes a disease management program for patients with coronary artery disease in a provincial health district with a population of 180,000. It discusses the rationale and methods behind the operationalisation of the main program elements, benefits achieved to date and challenges confronted. PMID- 11010584 TI - Use of monoclonal fluorescent antibodies for the detection of B. pertussis and B. parapertussis. AB - While it has been said that PCR is likely to replace DFA as the test of choice, accessibility to and cost of the technique may limit the number of laboratories that are able to implement the process. Still, the advantage of DFA staining relative to other diagnostic methods, including PCR, is the provision of a rapid result and lower overall cost. The laboratory diagnosis of pertussis remains problematic, and polyclonal DFA reagents for its detection have brought into question the utility of DFA. However, the integration of a commercially available monoclonal DFA reagent, in combination with the development and standardization of existing procedures, should provide clinicians with an improved method for the diagnosis and epidemiology of pertussis. PMID- 11010585 TI - Laboratory support in a regional approach to joint purchasing and materials management. PMID- 11010586 TI - Evaluation of a Troponin I assay employing enhanced chemiluminescence technology. PMID- 11010587 TI - Rapid identification of Haemophilus and other bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. PMID- 11010588 TI - Deconstructing disease: a pathologist's perspective. PMID- 11010589 TI - Cost-effective liquid-fixed cytology specimen processing with an economical cytocentrifuge. PMID- 11010590 TI - Comparison of metal ions in biological samples of schizophrenic patients and control subjects. PMID- 11010591 TI - Polystyrene microsphere spray standards based on CFC-free inhaler technology. AB - Metered pressurized sprays were formulated containing polystyrene microspheres (PSM) suspended in 5% w/w ethanol in 1,1,1,2-tetrafluoroethane (HFA 134a). Suspensions with defined number concentrations, packaged in pressure-resistant glass containers, and fitted with specified 50-microL metering values and actuators were sonicated and fired (as if they were metered dose inhalers or MDIs). Following propellant evaporation, PSM dispersions in air containing > 98% singlets resulted from actuation of 3, 5, and 8 microns PSM suspensions containing 0.00125% w/w 3 microns, 0.05% w/w 5 microns or 0.3% w/w 8 microns PSMs, respectively. Spray characteristics from these systems depended on PSM concentration, PSM size, PSM source, and actuator dimensions. Adhesion of suspended PSMs to the internal glass surfaces of the containers were easily reversed by sonication but not prevented by a wide range of surfactants. Over a period of 6 months storage, these formulations produced reproducible PSM aerosols with known aerodynamic properties useful for calibration purposes. PMID- 11010592 TI - Aerosol morphometry and aerosol bolus dispersion in patients with CT-determined combined pulmonary emphysema and lung fibrosis. AB - The simultaneous occurrence of pulmonary fibrosis and emphysema may present considerable problems in clinical assessment. Recent studies have shown that Aerosol Derived Airway Morphometry (ADAM) and Aerosol Bolus Dispersion (ABD) are changed in patients with pulmonary emphysema. This study was performed to assess the effect of simultaneous lung fibrosis in patients with emphysema on ADAM and ABD. ADAM and ABD measurements were performed in 20 patients with lone high resolution CT scan (HRCT) confirmed emphysema (E), and compared to those in 15 emphysematics with HRCT-confirmed superimposed pulmonary fibrosis (FE). In both groups the peripheral effective airspace dimension (EAD) (E: 0.63 +/- 0.20 mm; FE: 0.60 +/- 0.27 mm, N.S.) was increased by more than a factor of two compared to that of healthy subjects (0.28 +/- 0.05 mm) (p < 0.001). Patients with E showed a significantly higher bolus dispersion than patients with FE (724 +/- 122 cm3 vs. 546 +/- 80 cm3; p < 0.001). However, in patients with FE, bolus dispersion was still significantly higher than in previously published control groups of healthy subjects (546 +/- 80 cm3 vs. 455 +/- 68 cm3; p < 0.001). The results of this study confirm that ADAM and ABD are powerful tools for identifying emphysema even in patients with superimposed pulmonary fibrosis. PMID- 11010593 TI - Characterization of the laryngeal jet using phase Doppler interferometry. AB - The purpose of this study was to characterize the effects of the laryngeal jet on inhalation air flows in the trachea, and to extend these ideas to further an understanding of aerosol deposition in this region. Phase Doppler Interferometry was used to characterize axial velocity and turbulence intensity contours in the tracheal section of a cadaver-based larynx-trachea model. An array of 30 measurements was made at each of 6 downstream planes within the tracheal portion of the model (immediately downstream of the larynx, and at 0.5, 1, 2, 3, and 4 diameters further downstream). The flow was characterized for steady state flow at three Reynolds numbers (1250, 1700, and 2800). The Re = 1250 case approximates the inhalation of a 6-year-old child. Reverse flows with significant velocities were noted in the anterior trachea within one diameter downstream of the larynx, for all three flow cases. The cross sectional area of the reverse flow regions was larger for the lower Reynolds number cases. These reverse flows are a consequence of the nearly triangular shape of the lumen between the vocal folds in the larynx and constitute a potential deposition mechanism. High levels of axial turbulence intensity were noted near the anterior/left tracheal walls within one diameter downstream of the larynx. This indicates the potential for deposition due to turbulence in this region. Turbulence levels were still significant after four downstream diameters, indicating the potential for turbulent deposition at positions further downstream, including the bronchial tree where passage diameters are smaller. Contrary to expectations, turbulence levels were approximately 20% higher for the Re = 1250 case compared to the Re = 2800 at the furthest downstream locations (with 99% confidence). This is likely due to the complex nature of the confined jet flow. PMID- 11010594 TI - Validation of a new breathing simulator generating and measuring inhaled aerosol with adult breathing patterns. AB - The use of breathing simulators for the in vitro determination of the inhaled mass of drug from nebulizers has become widely accepted. Their use is, however, based on the assumption that there is a correlation between the in vitro and in vivo inhaled mass of drug. The aim of the study was therefore to investigate whether a new breathing simulator--the MIMIC Breathing Emulator (Medic-Aid Limited, Bognor Regis, UK)--could accurately emulate the in vivo inhaled mass of budesonide suspension for nebulization. Eight adult healthy subjects were included. Each subject inhaled for 2 min from a Spira Module 1 jet nebulizer (Respiratory Care Center, Hameenlinna, Finland), charged with 1.0 mg of budesonide suspension for nebulization (0.5 mg mL-1, 2 mL suspension, AstraZeneca, Sweden) and supplied with an inhaled mass filter between the nebulizer and the subject. The breathing patterns were recorded during the nebulization and simulated in vitro at two different experimental sites (simulations A and B) with the breathing simulator. With the patients breathing through the filters (in vivo test), inhaled mass of budesonide averaged 103.6 micrograms. The two in vitro experiments using the simulator revealed similar results with in vitro simulation A equal to 101.0 micrograms and simulation B 99.1 micrograms. There were no statistically significant differences between the in vivo results and those of in vitro simulation A. Results were significantly different for simulation B (p = 0.032) although the difference was less than 4.5%. These data indicate that the breathing simulator can be used to accurately simulate sine waveforms, human breathing patterns, and the in vitro and in vivo inhaled mass of budesonide suspension for nebulization. PMID- 11010595 TI - The choice of compressor effects the aerosol parameters and the delivery of tobramycin from a single model nebulizer. AB - Recent U.S. Phase III trials of the aerosolized delivery of tobramycin to cystic fibrosis (CF) patients demonstrated a significant improvement in pulmonary function and in sputum bacterial density. These trials used the Pari LC Plus nebulizer and DeVilbiss Pulmo-Aide compressor. This compressor is not generally available in Europe, and its power requirements do not match the European power supply. Thus alternate compressors were evaluated, using the LC Plus nebulizer, in preparation for European clinical trials. Aerosol particle size distribution, nebulization time (min), and the respirable dose of tobramycin (mg within 1-5 mu) were obtained for seven compressor models. The respirable quantity delivered by each of the European compressors (240 Volts, 50 Hz) was compared to the LC Plus and PulmoAide compressor (120 Volts, at 60 Hz). The U.S. system delivered 71.4 mg of the 300 mg instilled dose within the respirable range; using the European compressors, between 63.0 and 74.8 mg was delivered. With a 97% confidence that the delivered tobramycin was within 20% of the standard, we conclude that the SystAm 23ST, MedicAid CR50 and CR60, Pari Master and the Pari Boy compressors are equivalent to the U.S. standard; the Hercules and the SystAm 26ST compressors were not statistically equivalent to the standard. Using the LC Plus nebulizer, five European compressors delivered doses of TOBI that are similar to the doses delivered by the DeVilbiss PulmoAide compressors, and thus may be expected to produce clinical results similar to those of the U.S. trials. PMID- 11010596 TI - Comparing HFA MDI aerosol delivery in spacers. PMID- 11010597 TI - Regional mucociliary clearance in patients with cystic fibrosis. AB - This paper reports on a large retrospective analysis of mucociliary clearance (MCC) studies in a group of 59 patients with cystic fibrosis (CF) and 17 age matched healthy subjects. As many of the CF patients were studied on multiple occasions, a total of 184 patient studies are presented. MCC was measured using a radioaerosol and gamma camera technique. In addition to whole lung clearance, MCC was measured from the central, intermediate, peripheral, basal, mid and apical regions of the lung. MCC was markedly decreased in the CF patient group. Not only was whole lung clearance (14.2 +/- 1.4% vs. 28.0 +/- 3.7%) impaired, but also clearance from the central (19.1 +/- 1.9% vs. 35.6 +/- 4.3%), intermediate (10.7 +/- 1.6% vs. 25.5 +/- 3.7%), apical (12.4 +/- 2.6% vs. 31.6 +/- 4.6%) and mid (14.0 +/- 1.9% vs. 30.4 +/- 4.0%) regions. Attempts were made to identify factors that may have influenced MCC in both the normal subjects and CF patients. Age, gender, body mass index, patient genotype, penetration index, spontaneous cough, and various lung function parameters were entered into a stepwise multiple regression model, but none of the factors proved to be statistically important in determining MCC. Both intrasubject repeatability and intersubject variability estimates are presented for the patients and normal subjects that had multiple studies. The values were found to be remarkably similar for both CF patients and normal subjects and for both intra- and intersubject repeatability. With marked deviation from normal ranges and good repeatability, the measurement of MCC in CF patients would seem to be a valuable outcome measure for clinical trials involving new pharmaceuticals and physical therapy designed to improve removal of secretions from the airways. PMID- 11010598 TI - Effects of dextran sulfate on tracheal mucociliary velocity in dogs. AB - We have shown that low molecular weight dextran, as a potential mucolytic agent, reduced the viscoelasticity and spinnability of cystic fibrosis (CF) sputum and improved its ciliary transportability in vitro; it also reduced viscoelasticity of healthy dog mucus in in vitro testing. In anesthetized dogs, dextran administered by aerosol at 65 mg/mL increased tracheal mucus velocity, but this increase was not sustained for higher concentrations. The purpose of the present study is to evaluate whether low mol. wt. dextran sulfate, a charged oligosaccharide, exhibits similar effects to previously tested neutral dextran when administered by aerosol to anesthetized dogs in terms of mucus rheology and mucociliary clearance rate. Healthy mongrel dogs were anesthetized with pentobarbital and intubated. Aerosols of Ringer's solution or dextran sulfate (m.w. 5000) dissolved in Ringer's were generated by Pari LC STAR nebulizer, and delivered during 30-min periods of spontaneous breathing. Tracheal transepithelial potential difference (PD, using agar filled electrodes) and tracheal mucociliary velocity (TMV, by charcoal marker particle transport) were measured under bronchoscopic control, and mucus for viscoelasticity analysis by magnetic rheometry was collected by the endotracheal tube method. We performed experiments in seven dogs, involving 30-min administrations of aerosol, separated by 30-min periods of no aerosol. All dogs received inhalations of 6.5 mg/mL, 20 mg/mL, and 65 mg/mL dextran sulfate. Tracheal mucus viscoelasticity (average log G* over 1-100 rad/s) decreased progressively with increasing dose of dextran sulfate; for the highest concentration (65 mg/mL), log G* decreased by a factor of 2.61 (p = 0.021). A modest increase in the TMV was observed for the first dose of dextran sulfate (128% of baseline at 6.5 mg/mL, p = 0.066); thereafter TMV was stable. PD increased significantly at each concentration of dextran sulfate compared with Ringer control; however, there was no additional change between the three groups. The solids content of collected airway fluid (%SC) was gradually increased during successive 30-min dextran sulfate aerosols, indicating a significant residence time for the dextran in the mucus, and correlating with the decrease in viscoelasticity. These results suggest that dextran sulfate may be potentially of therapeutic value as a mucolytic agent, assisting mucus clearance by cough and physiotherapy, although whether it stimulates mucociliary clearance remains to be proven. PMID- 11010599 TI - Pulmonary drug delivery from the Taifun dry powder inhaler is relatively independent of the patient's inspiratory effort. AB - The Taifun dry powder inhaler (Leiras OY, Turku, Finland) is a breath-actuated, multidose device, each metered dose containing 200 micrograms of budesonide. A two-way randomized crossover gamma scintigraphic study was performed in 10 asthmatic patients to determine the in vivo deposition pattern of budesonide inhaled from the Taifun. In vitro radiolabelling validation studies demonstrated that the radiolabel could be used as an accurate marker to assess in vivo drug deposition. Patients used either maximal inspiratory effort (targeted peak inhalation flow 30 L/min) or submaximal inspiratory effort (targeted peak inhalation flow 15 L/min) on each study day. Mean (S.D.) whole lung deposition (% of metered dose) was 34.3 (5.8)% and 29.6 (5.9)% for the two inhalation flows. The intersubject coefficient of variation in lung deposition was less than 20% on both study days. Drug was deposited uniformly across the central, intermediate, and peripheral lung regions for maximal and submaximal inspiratory efforts. The study suggests that the Taifun is a superior drug delivery device compared with many other inhalers, in terms of the amount of drug deposited in the lungs, the reproducibility of the lung dose, and the relative flow--independence of lung deposition. PMID- 11010600 TI - Introduction to the pharmacoeconomics of herbal medicines. AB - This article explores the need to perform pharmacoeconomic evaluations of herbal medicines and assesses the extent to which this approach has been applied so far to these products. There seems to be no compelling need for pharmacoeconomic analyses of herbal over-the-counter medicines, but such analyses are certainly warranted for herbal prescription medicines that have a high level of reimbursement. Such preparations are used in Germany, in particular, where physicians prescribed ginkgo, hawthorn, St John's wort, horse-chestnut and saw palmetto to a value of more than DM50 million each in 1996. In our survey, only a single pharmacoeconomic study, of uncertain quality, was found on these 5 herbs, whereas several pharmacoeconomic reports on synthetic competitors were retrieved. The time has come to submit highly reimbursed herbal prescription medicines to the same rigorous pharmacoeconomic evaluations as their synthetic competitors. At present, such studies are particularly important for Germany, but in the future they may also become relevant for other countries, inside as well as outside Europe. PMID- 11010601 TI - Social phobia. Epidemiology and cost of illness. AB - Social phobia is a prevalent and debilitating psychiatric disorder. It entails significant economic costs in the form of educational underachievement, increased financial dependency, decreased work productivity, social impairment and poorer quality of life. It is associated with increased prevalence of other psychiatric disorders including depression and alcohol dependence. Its onset is early and typically precedes onset of comorbid disorders. Despite its debilitating effects, social phobia is often unrecognised and is undertreated. The availability of efficacious pharmacological and psychotherapeutic treatments for social phobia makes it imperative to carefully consider the impact of this disorder and to consider the cost effectiveness of available treatments. PMID- 11010602 TI - A review of alternative approaches to healthcare resource allocation. AB - The resources available for healthcare are limited compared with demand, if not need, and all healthcare systems, regardless of their financing and organisation, employ mechanisms to ration or prioritise finite healthcare resources. This paper reviews alternative approaches that can be used to allocate healthcare resources. It discusses the problems encountered when allocating healthcare resources according to free market principles. It then proceeds to discuss the advantages and disadvantages of alternative resource allocation approaches that can be applied to public health systems. These include: (i) approaches based on the concept of meeting the needs of the population to maximising its capacity to benefit from interventions; (ii) economic approaches that identify the most efficient allocation of resources with the view of maximising health benefits or other measures of social welfare; (iii) approaches that seek to ration healthcare by age; and (iv) approaches that resolve resource allocation disputes through debate and bargaining. At present, there appears to be no consensus about the relative importance of the potentially conflicting principles that can be used to guide resource allocation decisions. It is concluded that whatever shape tomorrow's health service takes, the requirement to make equitable and efficient use of finite healthcare resources will remain. PMID- 11010603 TI - Long-term treatment of multiple sclerosis with interferon-beta may be cost effective. AB - Multiple sclerosis (MS) is a devastating disease that can occur in early life, progressing to rapid disability and loss of physical, psychosocial and economic functioning, significantly affecting quality of life. The traditional treatment for MS has been symptomatic, treating acute relapses without affecting the underlying disease. The introduction of interferon-beta (IFN beta) has offered significant clinical benefits by reducing the frequency of relapses and slowing disease progression. Although the costs of this treatment are high, the costs to society of caring for a patient disabled by MS are greater, and if IFN beta can delay disease progression in the longer term, the economic impact would be substantial. Previous pharmacoeconomic studies of IFN beta have suggested that benefits can only be achieved at extremely high cost, with reported cost effectiveness measures of up to 1 million pounds sterling (Pound) per quality adjusted life year (QALY) [1995 values]. However, these studies have considered only the short term benefits of IFN beta treatment: over 2 to 3 years, the impact of treatment on patients' quality of life is relatively small, and cost-utility analyses that do not consider longer term benefits nor include societal costs may be misleading. The model reported here is based on the hypothesis that the delay in disease progression seen in short term clinical trials is likely to continue if treatment is continued. The model also assumes that the delay in disease progression, which represents a reduction in brain atrophy, will result in lasting clinical benefits even if treatment is stopped. These assumptions are strongly supported by clinical trial data and the treatment hypothesis itself. A delay in disease progression will result in a significant improvement in functioning and quality of life, and if the costs associated with increased disability can be postponed, even long term treatment of MS with IFN beta can be shown to be cost effective. Using resource utilisation costs derived from an economic evaluation of MS in the UK, it was possible to calculate the impact of delaying disease progression in terms of both health service and societal costs. An estimate of mean disease progression in patients with MS treated with IFN beta 1a compared with patients who did not receive disease-modifying agents suggested that significant cost savings would be realised after about 12 years' treatment with IFN beta-1a. The application of utility scores to the disease progression curves also facilitated estimates of cost effectiveness, with cost per QALY values ranging from 27,036 Pounds after 2 years' treatment with IFN beta-1a to 37,845 Pounds after 20 years' treatment (1995 values). PMID- 11010604 TI - Using economic evaluations to make formulary coverage decisions. So much for guidelines. AB - BACKGROUND: It is mandatory for drug manufacturers requesting formulary inclusion under the British Columbia (BC) provincial drug plan to submit a pharmacoeconomic analysis according to published guidelines. These submissions are reviewed by the Pharmacoeconomic Initiative (PI) of BC. OBJECTIVE: To assess the compliance of submitted studies with specific criteria outlined in the guidelines, to assess the methodological quality of individual submissions, and to demonstrate the importance of submitting guidelines-compliant pharmacoeconomic analyses. DATA AND METHODS: All submissions between January 1996 and April 1999 assessed by the PI of BC were included. Submissions were reviewed according to a checklist to establish compliance with respect to choice of comparator drug, study perspective, sensitivity analysis, analytical horizon and discounting. Submissions were examined for association between analytical technique and author, and between source of submission and compliance. Association between compliance and recommendation for approval was also examined. RESULTS: 95 applications were reviewed. Seven submitted no analyses. There were 25 cost comparison/consequence, 14 cost-effectiveness, 11 cost-minimisation, 9 cost utility/benefit and 29 budget-impact analyses. 65 of these 88 submissions failed to comply with guidelines. Of these, 45% used an inappropriate comparator drug, 61% lacked a sensitivity analysis, 73% used a third-party payer and excluded a societal perspective, 66% did not provide a long term evaluation and 25% did not specify any time horizon. 80% of noncompliant studies were cost comparison/consequence or budget-impact analyses (p < 0.001, Fisher's Exact). Of 25 cost-comparison/consequence and 29 budget-impact analyses, 19 (76%) and 24 (83%), respectively, were industry-conducted, whereas cost-effectiveness (11 of 14) and cost-utility/benefit (6 of 9) analyses were mostly subcontracted to private consultants or academics (p < 0.001, Fisher's Exact). 74% of all submissions (compliant and noncompliant) were not recommended by the PI for listing as a provincial drug plan benefit, 16% received approval for restricted benefit and 9% were recommended as full benefit. 80% of the noncompliant submissions were not recommended (p = 0.06, Fisher's Exact test). Moreover, a strong association between type of analysis and type of recommendation was found (p = 0.03, Fisher's Exact test). Cost-comparison/consequence and budget-impact analyses were less likely to be recommended. IMPLICATIONS OF FINDINGS: Our findings show poor compliance with guidelines, especially among industry conducted studies. Possible explanations are lack of expertise in pharmacoeconomics and/or scepticism regarding the importance of guidelines and submission quality in decision making. As corroborated by the strong associations between type of recommendation and compliance, and between type of recommendation and type of analysis, these 2 characteristics have a significant impact on decision making. PMID- 11010605 TI - Health-related quality of life in asymptomatic patients with HIV. Evaluation of the SF-36 health survey in Italian patients. AB - OBJECTIVE: To investigate the psychometric performance and clinical validity of the 36-Item Short Form (SF-36) health survey when completed by asymptomatic HIV positive Italian patients and to compare their health profile with a representative sample of 2031 Italian citizens (the Italian norm). PATIENTS AND METHODS: This was an observational, multicentre, cross-sectional survey. Microbiologists throughout Italy recruited asymptomatic HIV-positive individuals who were aged at least 18 years and aware of their infection. Investigators collected demographic, social, clinical and treatment data. Patients, classified into 2 clinical categories (A1 and A2) according to explicit pre-defined criteria, completed the SF-36 health survey in the context of a medical visit. RESULTS: Between April and July 1996, 46 microbiologists recruited 214 patients (201 evaluable). No inconsistent responses were observed in 96% of the sample. The usually recommended psychometric standards were satisfied, and the internal consistency reliability indices were always greater than 0.70. Weak to moderate associations were found between SF-36 health survey scores and physicians' estimates of patients' physical performance, while no significant associations were found with CD4+ counts. On average, HIV-positive patients reported lower scores than the Italian norm, and patients in category A2 showed lower scores than patients in A1. These differences were more relevant in scales describing role limitations, general health perception, and psychological well-being. CONCLUSION: Our study showed that the SF-36 health survey maintained its psychometric properties in a sample of Italian asymptomatic HIV-positive patients and produced data that showed its validity and robustness in such a setting. PMID- 11010606 TI - The impact of pituitary adenoma on morbidity. Increased sick leave and disability retirement in a cross-sectional analysis of Swedish national data. AB - OBJECTIVE: To quantify sick leave and medical retirement for the year 1989 in adults with a history of nonsecreting pituitary adenoma. DESIGN AND SETTING: A cross-sectional analysis of official Swedish data from 1989 was performed. Sick leave and disability pension data for these patients were obtained from the Swedish National Social Insurance Board, which also supplied information on sick leave taken by an age- and gender-matched control population of 5121 individuals. Uptake of disability pensions and the reasons for drawing these pensions in the study group were compared with national statistics. STUDY POPULATION: A group of 809 eligible adults with nonsecreting pituitary adenoma was identified from the national cancer registry. MAIN OUTCOME MEASURES AND RESULTS: Almost a quarter of the patients with a history of pituitary adenoma had retired due to ill health (23.8%); this was twice the figure expected from national statistics (11.5%). Similar results were obtained when men and women were considered separately. Some patients (17%) had received disability pensions before the diagnosis of pituitary adenoma, but the majority (75%) retired on medical grounds at least 1 year after their diagnosis. The reasons for early retirement in the study group were largely related to the diagnosis of pituitary adenoma (e.g. neoplasm, endocrine disorders), but there was some evidence of an increase in the number of disability pensions awarded because of diseases of the nervous system and sensory organs. Patients with a history of pituitary adenoma took significantly more sick leave in 1989 than those in the control group (mean 40.2 vs 24.0 days), and this significant difference applied to both sexes. CONCLUSIONS: A cross-sectional analysis of Swedish national data from 1989 showed excess morbidity, relative to reference data, in patients with a history of nonsecreting pituitary adenoma. Suboptimal conventional hormone replacement therapy and untreated growth hormone deficiency might to some extent explain the increased morbidity found in the present study. PMID- 11010607 TI - Economic evaluation of enoxaparin sodium versus heparin in unstable angina. A French sub-study of the ESSENCE trial. AB - OBJECTIVES: To perform an evaluation from the societal perspective of the cost of treatment with enoxaparin sodium versus unfractionated heparin (UFH) in patients with unstable angina and non-Q wave myocardial infarction in France. DESIGN: Four complementary cost-minimisation analyses based on the results of the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q wave Coronary Events (ESSENCE) international trial were conducted. We assessed differences in medical resource consumption and in duration of hospital stay in the whole study population (n = 3171) and for the French patients (n = 133). RESULTS: Results were consistent for the study group as a whole and for the French subgroup. Among patients treated with enoxaparin sodium, there was a statistically significant reduction in the use of angiography and percutaneous transluminal coronary angioplasty (whole group study: p = 0.024 and 0.006, respectively) and a trend towards shorter lengths of hospital stay. The differences in angiography and angioplasty rates led to estimated average net cost savings with enoxaparin sodium of French Francs (FF)1555 per treated patient (whole study population) and FF9993 (French subgroup) [1996 values]. The analyses based on the duration of hospital stay resulted in estimated net cost savings with enoxaparin sodium of between FF1014 per treated patient (whole study population) and FF2804 (French subgroup). CONCLUSION: Our study confirmed earlier results which show that enoxaparin sodium is cost saving in the treatment of unstable angina. PMID- 11010609 TI - Is universal coverage the American way? PMID- 11010608 TI - Economic issues in the prevention of vertical transmission of HIV. AB - In the absence of interventions, 20% of infants born to women infected with HIV acquire infection from their mother at or before delivery. A further 15% are infected through breast feeding. Prenatal testing for HIV allows infected women to be reliably identified so that they can receive antiretroviral therapy and, in countries with safe water supplies, be advised not to breast feed. These and other interventions can reduce the risk of transmission to 5% or less. Economic evaluations of prenatal testing for HIV are reviewed and compared in this article, and future research priorities outlined. These studies set the costs of testing and intervention against the averted lifetime costs of paediatric infection, and generate estimates of the HIV prevalence threshold above which there would be a net cost saving, or calculate the cost per life-year saved given a particular prevalence. In the developed world, prenatal testing has been adopted in many countries, and recent economic analyses broadly support this. Future research is likely to focus on the incremental benefits of different antiretroviral regimens in lowering transmission rates still further, with or without elective caesarean section, and the possibility that some may lead to adverse effects in uninfected infants exposed to them in utero. Some earlier assessments in resource-poor settings concluded that prenatal testing was unaffordable or of doubtful cost effectiveness. This negative conclusion appears to be the result of very low estimates of the lifetime costs of paediatric HIV infection, together with developed world conceptions of pre-test counselling. The demonstration that nevirapine reduces transmission risk at a low cost has transformed the outlook, and there is hope that antiretrovirals can act prophylactically to prevent infection of the breast-fed child. However, to achieve a sustained reduction in vertical transmission there may be a need to evaluate the need for a strengthened infrastructure to deliver prenatal HIV testing and treatment, as well as programmes to reduce HIV incidence in adults. PMID- 11010611 TI - 'Problem patients': a fresh look at an old vexation. PMID- 11010610 TI - Finding success in a capitated environment. PMID- 11010612 TI - High-volume practice: are there trade-offs? PMID- 11010613 TI - Understanding confidentiality and nonsolicitation clauses. PMID- 11010614 TI - Local innovations provide managed care for the uninsured. AB - A number of communities in the United States are adopting a managed care approach to caring for low-income uninsured individuals. This Issue Brief focuses on such programs in five of the 12 communities that the Center for Studying Health System Change (HSC) is tracking intensively. It describes the local market factors that motivated the creation and varying design of these initiatives, all of which seek to increase access to primary and preventive care while managing the use of more costly inpatient and emergency care. The Issue Brief also discusses the long term viability of these programs as they attempt to simultaneously expand access to services and contain costs for this growing population. PMID- 11010615 TI - Who has a choice of health plans? AB - Policy makers are concerned that consumers have no voice in the changing health care system. They debate, however, whether the consumers' voice should be heard through regulation, such as patient protections, or the marketplace. For market forces to work in the consumers' interest, consumers must have a choice of plans and detailed information on which to base their choice. New survey data from the Center for Studying Health System Change (HSC) suggest that more consumers have a choice of plans than is generally believed, and that the proportion of consumers who have plan choice is increasing. According to HSC's 1998-1999 Household Survey, 64 percent of families have a choice of health plans--two percentage points higher than two years ago. PMID- 11010616 TI - Improving quality and preventing error in medical practice. PMID- 11010617 TI - Selecting a range of options. PMID- 11010618 TI - Laparoscopic Nissen fundoplication. PMID- 11010619 TI - Rheumatology. PMID- 11010620 TI - Dispelling the myths and stigma of mental illness: the Surgeon General's report on mental health. PMID- 11010621 TI - Regulations on statements made for dietary supplements concerning the effect of the product on the structure or function of the body. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing final regulations defining the types of statements that can be made concerning the effect of a dietary supplement on the structure or function of the body. The regulations also establish criteria for determining when a statement about a dietary supplement is a claim to diagnose, cure, mitigate, treat, or prevent disease. This action is intended to clarify the types of claims that may be made for dietary supplements without prior review by FDA and the types of claims that require prior authorization as health claims or prior approval as drug claims. PMID- 11010622 TI - Medical devices; laser fluorescence caries detection device. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the laser fluorescence caries detection device into class II (special controls). The special controls that will apply to this device are set forth below. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. PMID- 11010623 TI - Hematology and pathology devices; reclassification; restricted devices; OTC test sample collection systems for drugs of abuse testing. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is reclassifying over-the-counter (OTC) test sample collection systems for drugs of abuse testing from class III (premarket approval) into class I (general controls) and exempting them from premarket notification (510(k)) and current good manufacturing practice (CGMP) requirements. FDA is also designating OTC test sample collection systems for drugs of abuse testing as restricted devices under the Federal Food, Drug, and Cosmetic Act (the act) and establishing restrictions intended to assure consumers that: The underlying laboratory test(s) are accurate and reliable; the laboratory performing the test(s) has adequate expertise and competency; and the product has adequate labeling and methods of communicating test results to consumers. Finally, FDA is adding a conforming amendment to the existing classification regulation for specimen transport and storage containers to clarify that it does not apply to specimen transport and storage containers that are part of an OTC test sample collection system for the purpose of testing for the presence of drugs of abuse or their metabolites in a laboratory. PMID- 11010624 TI - Medical devices; gastroenterology-urology devices; nonimplanted, peripheral electrical continence device. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the nonimplanted, peripheral electrical continence device into class II (special controls). The special controls that will apply to this device are set forth below. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. PMID- 11010625 TI - Over-the-counter human drugs; labeling requirements; final rule; technical amendment. Food and Drug Administration, HHS. Final rule; technical amendment. AB - The Food and Drug Administration (FDA) is amending the regulation that established a standardized format and standardized content requirements for the labeling of over-the-counter (OTC) drug products, and is amending several related OTC drug product labeling regulations. This amendment corrects and conforms several aspects of the new labeling requirements to other regulatory provisions and eliminates unnecessary text from the new labeling regulation. PMID- 11010626 TI - Diagnostic criteria for traumatic grief. AB - This article reviews the rationale for the development of diagnostic criteria for Traumatic Grief. Traumatic Grief is a new nosologic entity that a panel of experts recently proposed. It is a direct descendent of the concept of pathologic grief, and it has roots in attachment behavior, separation distress, and traumatic distress. We present consensus, diagnostic criteria for Traumatic Grief and discuss them in relation to another recently proposed set of criteria. In conclusion, we recommend the development of empirically tested, consensus, diagnostic criteria for Traumatic Grief. Diagnostic criteria would facilitate early detection and intervention for those bereaved persons afflicted by this disorder and lead to additional studies of the prevalence, the nature, and the treatment of the disorder. PMID- 11010627 TI - Relationship of recollections of first death experience to current death attitudes. AB - Previous research (Dickinson, 1992) has investigated adults' memories of their first childhood experience with death. The present study extended this work to examine the relationship of various qualitative aspects of the first experience with death to current death attitudes. College students (196 females, 101 males) described their first childhood death experience and provided memories of: (a) who died; (b) whether parental discussion of death had occurred; (c) whether they had unanswered questions at the time; and (d) whether they had cried at the time. They then completed the Death Attitude Profile-Revised (DAP-R; Wong, Reker, & Gesser, 1994). Results indicated that although boys and girls did not respond differently to the first death experience, gender did appear to moderate the relationship between the qualities of this experience and current death attitudes. PMID- 11010628 TI - Blues fans and suicide acceptability. AB - Research has neglected the possible impact of the blues music subculture on suicide acceptability (SA). The sad themes in the blues may attract suicidal persons and reinforce their suicidal moods and attitudes. The present study performs the first test of the thesis that associates SA with being a blues fan. It uses data on a national sample of 961 adults drawn from the General Social Survey of 1993. The results of a multivariate logistic regression analysis found that blues fans were no more accepting of suicide than nonfans. However, blues fanship was found to have substantial indirect effects on SA through its influence on such factors as lowered religiosity levels, the most important predictor of SA. Race-specific analyses found more support for the model for whites than for African Americans. PMID- 11010629 TI - Making decisions about tube feeding for severely demented patients at the end of life: clinical, legal, and ethical considerations. AB - Caregivers and family members are forced to deal with questions about tube feeding at the end of life for hundreds of thousands of patients suffering from severe dementia every year. But decisions about accepting or forgoing artificial nutrition and hydration (ANH) tend to be made in haste, late in the game, without benefit of full information. Oftentimes, this leads to increased patient suffering and the inefficient use of medical resources. Surviving family members and caregivers may experience a haunting sense of guilt, wondering if they made the right decision at the right time. The professional literature suggests that forgoing ANH is an entirely appropriate alternative at the end of life. The vast majority of Americans say they do not want to be tube fed if mortally ill and are no longer able to eat on their own. Nonetheless, the provision of aggressive nutrition and hydration support for the severely demented population at the end of life is the norm in America. If there were more awareness of (a) the burdens typically associated with ANH; (b) the pathogenesis of terminal dehydration, and; (c) the medical, legal, and ethical acceptability of terminal dehydration, then forgoing of ANH might become more commonly considered. PMID- 11010630 TI - Cardiovascular, orthopedic, and physical medicine diagnostic devices; reclassification of cardiopulmonary bypass accessory equipment, goniometer device, and electrode cable devices. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is reclassifying from class I into class II the cardiopulmonary bypass accessory equipment device that involves an electrical connection to the patient, the goniometer device, and the electrode cable. FDA is also exempting these devices from the premarket notification requirements. FDA is reclassifying these devices on its own initiative based on new information. FDA is taking this action to establish sufficient regulatory controls that will provide reasonable assurance of the safety and effectiveness of these devices. PMID- 11010631 TI - From Chakrabarty to chimeras: the growing need for evolutionary biology in patent law. PMID- 11010632 TI - Gastroenterology-urology devices; effective date of requirement for premarket approval of the penile inflatable implant. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to require the filing of a premarket approval application (PMA) or a notice of completion of a product development protocol (PDP) for the penile inflatable implant, a generic type of medical device intended for the treatment of erectile dysfunction. This regulation reflects FDA's exercise of its discretion to require PMA's or PDP's for preamendments devices and is consistent with FDA's stated priorities and Congress' requirement that class III devices are to be regulated by FDA's premarket review. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. PMID- 11010634 TI - Medical devices; effective date of requirement for premarket approval for three preamendment class III devices. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to retain three class III preamendment devices in class III and to require the filing of a premarket approval application (PMA) or a notice of completion of a product development protocol (PDP) for the following devices: The lung water monitor, the powered vaginal muscle stimulator, and the stair-climbing wheelchair. The agency has summarized its findings regarding the degree of risk of illness or injury designed to be eliminated or reduced by requiring the devices to meet the statute's approval requirements and the benefits to the public from the use of the devices. PMID- 11010633 TI - Are insurance companies liable under the Americans with Disabilities Act? AB - Federal courts have split on the question of the applicability of the Americans with Disabilities Act to insurance coverage decisions that insurance companies make on the basis of disability; they have similarly split on other issues pertaining to the scope of that Act's application. In deciding whether to read the Act as prohibiting discrimination in insurance decisions that are often crucial in the lives of people with disabilities, courts have faced two problems. First, where it prohibits discrimination in the equal enjoyment of the goods and services of places of public accommodation, the Act's area of concern may be limited to the ability of people with disabilities to gain physical access to facilities; or that area may extend to all forms of disability-based discrimination in the provision of goods and services. This Comment argues that the language and legislative history of the Act are consistent only with the latter view. Second, the provision limiting the Act's applicability to insurance may create an exemption for all insurance decisions; or it may protect only the ability of an insurance company to make an insurance decision to the disadvantage of an insured with a disability where actuarial data support the decision. This comment argues that the ambiguous language of the limiting provision should be resolved in favor of the latter view. Legislative history and the broader background of the history of insurance discrimination law support this resolution. Consequently, the Act should be interpreted as prohibiting disability based discrimination by insurance companies in selling insurance policies and as defining discrimination as making disability-based insurance decisions without the support of actuarial data. By accepting this interpretation, courts can help stop the pattern of judicial narrowing of the Act's application through inappropriately restrictive statutory construction. PMID- 11010635 TI - Medical devices; reclassification and codification of the stainless steel suture. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is announcing that it has issued an order in the form of a letter to Alto Development Corp. (the petitioner) reclassifying the 316L stainless steel suture for use in abdominal wound closure, intestinal anastomosis, hernia repair, and sternal closure from class III (premarket approval) to class II (special controls). The order is being codified in the Code of Federal Regulations (CFR). Although FDA reclassified the device in 1986, it inadvertently neglected to publish a notice of the reclassification in the Federal Register or codify the change in the CFR. PMID- 11010636 TI - The mainstreaming of alternative medicine. PMID- 11010637 TI - New Jersey insurers expand cancer care coverage. PMID- 11010638 TI - Forgotten patients: the mentally ill. PMID- 11010639 TI - The crisis ahead in long-term care. PMID- 11010640 TI - Medical devices; reclassification and codification of the nonabsorbable expanded polytetrafluoroethylene surgical suture. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is announcing that it has issued an order in the form of a letter to W.L. Gore and Associates, Inc., reclassifying the nonabsorbable expanded polytetrafluoroethylene (ePTFE) surgical suture intended for use in soft tissue approximation and ligation, including cardiovascular surgery, from class III (premarket approval) to class II (special controls). Accordingly, the order is being codified in the Code of Federal Regulations (CFR). PMID- 11010641 TI - Nevada State plan; final approval determination. Occupational Safety and Health Administration (OSHA), U.S. Department of Labor. Final State plan approval- Nevada. AB - This document amends OSHA's regulations to reflect the Assistant Secretary's decision granting final approval to the Nevada State plan. As a result of this affirmative determination under section 18(e) of the Occupational Safety and Health Act of 1970, Federal OSHA's standards and enforcement authority no longer apply to occupational safety and health issues covered by the Nevada plan, and authority for Federal concurrent jurisdiction is relinquished. Federal enforcement jurisdiction is retained over any private sector maritime employment, private sector employers on Indian land, and any contractors or subcontractors on any Federal establishment where the land is exclusive Federal jurisdiction. Federal jurisdiction remains in effect with respect to Federal government employers and employees. Federal OSHA will also retain authority for coverage of the United States Postal Service (USPS), including USPS employees, contract employees, and contractor-operated facilities engaged in USPS mail operations. PMID- 11010642 TI - Determining disability. Railroad Retirement Board. Final rule. AB - The Railroad Retirement Board (Board) hereby amends its disability regulations to discontinue the current policy of conducting continuing disability reviews (CDR's) for medical recovery of disability annuitants in medical improvement not expected (MINE) cases. The Board has found that these reviews have not been cost effective and impose an unnecessary burden on the annuitant. PMID- 11010643 TI - Office of Inspector General; Medicare program; prospective payment system for hospital outpatient services. Health Care Financing Administration (HCFA), HHS, and Office of Inspector General (OIG), HHS. Final rule with comment period. AB - This final rule with comment period implements a prospective payment system for hospital outpatient services furnished to Medicare beneficiaries, as set forth in section 1833(t) of the Social Security Act. It also establishes requirements for provider departments and provider-based entities, and it implements section 9343(c) of the Omnibus Budget Reconciliation Act of 1986, which prohibits Medicare payment for nonphysician services furnished to a hospital outpatient by a provider or supplier other than a hospital, unless the services are furnished under an arrangement with the hospital. In addition, this rule establishes in regulations the extension of reductions in payment for costs of hospital outpatient services required by section 4522 of the Balanced Budget Act of 1997, as amended by section 201(k) of the Balanced Budget Refinement Act of 1999. PMID- 11010644 TI - Clinical chemistry devices; classification of the biotinidase test system. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the biotinidase test system into class II (special controls). The special control that will apply to this device is restriction to sale, distribution, and use as a prescription device. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying these devices into class II (special controls) in order to provide a reasonable assurance of the safety and effectiveness of the devices. PMID- 11010645 TI - Tax treatment of cafeteria plans. Internal Revenue Service (IRS), Treasury. Final regulations. AB - This document contains final regulations relating to section 125 cafeteria plans. The final regulations clarify the circumstances under which a section 125 cafeteria plan election may be changed. The final regulations permit an employer to allow a section 125 cafeteria plan participant to revoke an existing election and make a new election during a period of coverage for accident or health coverage or group-term life insurance coverage. PMID- 11010646 TI - Medicare program; coverage of, and payment for, paramedic intercept ambulance services. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule responds to public comments received on a final rule with comment period published on January 25, 1999 that implemented section 4531(c) of the Balanced Budget Act of 1997 concerning Medicare coverage of, and payment for, paramedic intercept ambulance services in rural communities. It also implements section 412 of the Medicare, Medicaid, and State Children's Health Insurance Programs Balanced Budget Refinement Act of 1999 by adding a new definition of a rural area. PMID- 11010647 TI - Revision of requirements applicable to albumin (human), plasma protein fraction (human), and immune globulin (human); confirmation in part and technical amendment. Food and Drug Administration, HHS. Direct final rule; confirmation in part and technical amendment. AB - The Food and Drug Administration (FDA) is confirming in part the direct final rule that appeared in the Federal Register of May 14, 1999 (64 FR 26282). The direct final rule amends the biologics regulations by removing, revising, or updating specific regulations applicable to blood derivative products to be more consistent with current practices and to remove unnecessary or outdated requirements. FDA is confirming the provisions for which no significant adverse comments were received. The agency received significant adverse comments on certain provisions and is hereby amending Title 21 Code of Federal Regulations to reinstate the former provisions. In addition, FDA is correcting the precision of the value for protein concentration that was inadvertently omitted from the codified section of the direct final rule. PMID- 11010648 TI - Federal old-age, survivors, and disability insurance and supplemental security income for the aged, blind, and disabled; evaluating opinion evidence. Social Security Administration. Final rules. AB - We are revising the Social Security and Supplemental Security Income (SSI) regulations concerning the evaluation of medical opinions to clarify how administrative law judges and the Appeals Council are to consider opinion evidence from State agency medical and psychological consultants, other program physicians and psychologists, and medical experts we consult in claims for disability benefits under titles II and XVI of the Social Security Act (the Act). We are also defining and clarifying several terms used in our regulations and deleting other terms. PMID- 11010649 TI - Medical devices; anesthesiology devices; classification of nitric oxide administration apparatus, nitric oxide analyzer, and nitrogen dioxide analyzer. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the nitric oxide administration apparatus, nitric oxide analyzer, and nitrogen dioxide analyzer into class II (special controls). The special control that will apply to these devices is a guidance document. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976 (the amendments), the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying these devices into class II (special controls) in order to provide a reasonable assurance of the safety and effectiveness of the devices. PMID- 11010651 TI - Medical devices; reclassification and codification of Neodymium:Yttrium:Aluminum:Garnet (Nd:YAG) Laser for peripheral iridotomy. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is announcing that it has issued an order in the form of a letter to Intelligent Surgical Lasers, Inc. (ISL), (now doing business as Escalon Medical Corporation), reclassifying the Neodymium:Yttrium:Aluminum:Garnet (Nd:YAG) Laser for use in peripheral iridotomy from class III to class II (special controls). Accordingly, the order is now being codified in the Code of Federal Regulations (CFR) as described below. PMID- 11010650 TI - Medical devices; exemptions from premarket notification; class II devices; vascular tunnelers. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is publishing an order granting in part a petition requesting exemption from the premarket notification requirements for vascular tunnelers with certain limitations. This rule will exempt from premarket notification stainless steel vascular tunnelers of single unit construction. FDA is publishing this order in accordance with procedures established by the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11010652 TI - Gastroenterology-urology devices: reclassification of the penile rigidity implant. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is reclassifying the penile rigidity implant from class III to class II when intended to provide penile rigidity in men diagnosed as having erectile dysfunction. The special control is the FDA guidance document entitled "Guidance for the Content of Premarket Notifications for Penile Rigidity Implants." This action is taken on FDA's own initiative based on new information. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the FDA Modernization Act of 1997. PMID- 11010653 TI - Medical device reporting: manufacturer reporting, importer reporting, user facility reporting, distributor reporting. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending its regulations governing reporting by manufacturers, importers, distributors and health care (user) facilities of adverse events related to medical devices. Amendments are being made to implement revisions to the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11010654 TI - Amendment of various device regulations to reflect current American Society for Testing and Material citations. Food and Drug Administration, HHS. Direct final rule. AB - The Food and Drug Administration (FDA) is amending certain references in various medical device regulations. The amendments update the references in those regulations to various standards of the American Society for Testing and Materials (ASTM) to reflect the current standards designations. Elsewhere in this issue of the Federal Register, FDA is publishing a companion proposed rule, under FDA's usual procedures for notice-and-comment, to provide a procedural framework to finalize the rule in the event that the agency receives any significant adverse comment and withdraws the direct final rule. PMID- 11010655 TI - Medical devices; exemption from premarket notification and reserved devices; class I. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending its classification regulations to designate class I devices that are exempt from the premarket notification requirements, subject to certain limitations, and to designate those class I devices that remain subject to premarket notification requirements under the new statutory criteria for premarket notification requirements. The devices FDA is designating as exempt do not include class I devices that have been previously exempted by regulation from the premarket notification requirements. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976 (the 1976 amendments), the Safe Medical Devices Act of 1990 (SMDA), and the FDA Modernization Act of 1997 (FDAMA). FDA is taking this action in order to implement a requirement of FDAMA. Elsewhere in this issue of the Federal Register, FDA is announcing that it is withdrawing proposed rules to revoke existing exemptions from premarket notification for two devices. PMID- 11010656 TI - New drug applications; drug master files. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revising its regulation governing drug master files (DMF's). FDA is removing the provision for submitting Type I DMF's and will no longer permit information submitted in a Type I DMF to be incorporated by reference in investigational new drug applications (IND's), new drug applications (NDA's), abbreviated new drug applications (ANDA's), or amendments or supplements to any of these. This rule is intended to eliminate submissions of information that are not necessary either to conduct inspections of manufacturing facilities or to review the chemistry, manufacturing, and controls sections of IND's, NDA's, and abbreviated applications. PMID- 11010657 TI - Per diem for nursing home care of veterans in State homes. Department of Veterans Affairs. Final rule. AB - This document amends regulations regarding the payment of per diem to State homes that provide nursing home care to eligible veterans. The intended effect of the final rule is to ensure that veterans receive high quality care in State homes. PMID- 11010658 TI - The myth of the replacement child: parents' stories and practices after perinatal death. AB - Parents bereaved by perinatal death adapt to their losses in different ways. When bereaved parents give birth to a child or children subsequent to a perinatal death, their constructions of the family necessarily change. The subsequent child is thought to be at risk of psychopathology (the replacement child syndrome) if parents have not sufficiently grieved their losses. This qualitative interview study examines the family stories told by bereaved parents, with particular attention to how parents represent the dead child and subsequent children in the current family structure. We categorized parents' stories as those which suggested that parents replaced the loss by an emphasis on parenting subsequent children, or maintained a connection to the dead child through storytelling and ritual behavior. The two ways in which parents maintained the connection were to preserve the space in the family that the dead child would have inhabited, or to create an on-going relationship with the dead child for themselves and their subsequent children. There seem to be multiple paths to parenting through bereavement. The place of rituals and memorial behavior is also examined. PMID- 11010659 TI - Does writing about the bereavement lessen grief following sudden, unintentional death? AB - Writing about traumatic events produces improvement in an array of areas including physical and psychological functioning. To see if these improvements extended to improved bereavement recovery after the accidental or homicidal death of a loved one, 64 undergraduates (51 women, 13 men) began, and 44 completed, a writing project. At pretest, they completed measures of depression, anxiety, grief, impact, and non-routine health visits. Then, they were randomly assigned to write about either the bereavement experience (profound condition), or innocuous topics (trivial condition). They wrote for 15 minutes a day for four days, then completed the same measures a second time (posttest). Six weeks later, they were mailed the same measures again (follow-up). A 2 (CONDITION: Profound versus Trivial) x 3 (Time: Pre-, Post-, or Follow-up) MANOVA yielded a significant main effect for time, but no main effect for condition and no interaction. Follow-up ANOVAs indicated that, across conditions, from pretest to follow-up testing participants reported less anxiety and depression, less impact, greater grief recovery, but about the same health center visits. A 2 (CONDITION) x 4 (Writing Day) MANOVA and follow-up tests indicated that those in the profound condition reported less subjective distress from Day 1 to Day 3, compared to those in the trivial condition. Combined with Kovac and Range (1999), present results suggest that writing projects may be more beneficial to those experiencing the unique bereavement of suicidal death, rather than those experiencing the nonintentional death of a loved one by accident or homicide. PMID- 11010660 TI - Hospice and assisted suicide: the structure and process of an inherent dilemma. AB - The hospice philosophy involves making terminal patients as comfortable as possible, empowering them with control of the time they have left, but neither hastening nor postponing death. The passage of Oregon's Death with Dignity Act in November of 1994, and the failure of the 1997 ballot measure to repeal it, made physician-assisted suicide another option for terminally ill people in that state, and focused increased attention on a conflict seemingly inherent in the hospice philosophy. We conducted interviews with 60 hospice providers, 43 in Oregon and, for comparison, 17 in the northeast, for their responses to this situation. The data reported here reflect some of the social and individual influences that come to bear as hospice providers attempt to resolve their dilemma. PMID- 11010661 TI - Shame and suicide: a case study. AB - Building on a recent paper by Lester (1997) on shame and suicide, a case is presented of a woman who made a serious attempt to kill herself, motivated in large part by shame. The therapeutic strategies employed are described, and the case brought to a successful resolution. PMID- 11010662 TI - Medicare and state health care programs: fraud and abuse; clarification of the initial OIG safe harbor provisions and establishment of additional safe harbor provisions under the anti-kickback statute. Office of Inspector General (OIG), HHS. Final rule. AB - This final rule serves both to add new safe harbor provisions under the Federal and State health care programs' anti-kickback statute, as authorized under section 14 of Public Law 100-93, the Medicare and Medicaid Patient and Program Protection Act of 1987, and to clarify various aspects of the original safe harbor provisions now codified in 42 CFR part 1001 (originally proposed in RIN 0991-AA74). Specifically, this final rule modifies the original set of final safe harbor provisions codified in 42 CFR 1001.952 to give greater clarity to that rulemaking's original intent. In addition, this final rule sets forth an expanded set of safe harbor provisions designed to protect additional payment and business practices from criminal prosecution or civil sanctions under the anti-kickback provisions of the statute. PMID- 11010663 TI - Medicare and Medicaid programs; programs of all-inclusive care for the elderly (PACE). Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This rule establishes requirements for Programs of All-inclusive Care for the Elderly (PACE) under Medicare and Medicaid. These are pre-paid, capitated programs for beneficiaries who meet special eligibility requirements and who elect to enroll. Programs must apply for approval and are evaluated in terms of specific criteria. Only a limited number of programs can be approved. Priority consideration will be given to applicants that have been operating under ongoing PACE demonstration projects. PMID- 11010664 TI - Medicare and Medicaid programs; religious nonmedical health care institutions and advance directives. Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This rule deletes all references to Christian Science sanatoria and sets forth the Medicare requirements for coverage and payment of services furnished by religious nonmedical health care institutions. It also sets forth the conditions of participation that religious nonmedical health care institutions must meet before they can participate in Medicare. It sets forth the methods we will use to pay religious nonmedical health care institutions and monitor expenditures for religious nonmedical health care institution services. Additionally, the rule presents the rules governing optional coverage of religious nonmedical health care institution services by States under the Medicaid program. PMID- 11010665 TI - Prescription Drug Marketing Act of 1987; Prescription Drug Amendments of 1992; policies, requirements, and administrative procedures. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to set forth procedures and requirements implementing the Prescription Drug Marketing Act of 1987 (PDMA), as modified by the Prescription Drug Amendments of 1992 (PDA) and the FDA Modernization Act of 1997 (the Modernization Act). The final rule sets forth requirements for the reimportation and wholesale distribution of prescription drugs; the sale, purchase, or trade of, or the offer to sell, purchase, or trade, prescription drugs that were purchased by hospitals or health care entities, or donated to charitable organizations; and the distribution of prescription drug samples. FDA is also amending certain sections of the regulations entitled "Guidelines for State Licensing of Wholesale Prescription Drug Distributors" to make them consistent with this final regulation. PMID- 11010666 TI - Establishment of a Medical Implant Communications Service in the 402-405 MHz band. Federal Communications Commission. Final rule. AB - This document establishes a Medical Implant Communications Service (MICS) operating in the 402-405 MHz band. MICS operations will consist of high-speed, ultra-low power, non-voice transmissions to and from implanted medical devices such as cardiac pacemakers and defibrillators. The rules will allow use of newly developed, life-saving medical technology without harming other users of the frequency band. PMID- 11010667 TI - It's only skin deep: FDA regulation of skin care cosmetics claims. PMID- 11010668 TI - The third wave of federal tort reform: protecting the public or pushing the constitutional envelope? PMID- 11010669 TI - Approval and promulgation of Hospital/Medical/Infectious Waste Incinerators: State Plan for designated facilities and pollutants: Idaho. Environmental Protection Agency (EPA). Direct final rule. AB - EPA is approving the State of Idaho's section 111(d) State Plan for controlling emissions from existing Hospital/Medical/Infectious Waste Incinerators (HMIWI). The plan was submitted on December 16, 1999, to fulfill the requirements of sections 111(d) and 129 of the Clean Air Act. The State Plan adopts and implements the Emissions Guidelines applicable to existing HMIWIs, and establishes emission limits and controls for sources constructed on or before June 20, 1996. EPA has determined that Idaho's State Plan meets CAA requirements and hereby approves this State Plan, thus making it federally enforceable. PMID- 11010670 TI - Placement of gamma-butyrolactone in List I of the Controlled Substances Act (21 U.S.C. 802(34)). Drug Enforcement Administration, Justice. Final rule. AB - Public Law 106-172, signed into law on February 18, 2000, and known as the "Hillory J. Farias and Samantha Reid Date-Rape Drug Prohibition Act of 1999," amends section 102(34) of the Controlled Substances Act as amended (CSA) by designating gamma-butyrolactone (GBL), the precursor to gamma-hydroxybutyric acid (GHB), as a List I chemical. Reflecting this change in stature, the Drug Enforcement Administration (DEA) is amending its regulation to reflect the status of GBL as a List I chemical subject to the requirements of the CSA and its regulations. Establishment of a threshold for GBL will be the subject of a separate rulemaking. Therefore, unless and until a threshold is established, any distribution of GBL is a regulated transaction as described by 21 CFR 1300.02(b)(28). All handlers of GBL must comply with the CSA regulatory requirements pertaining to List I chemicals as described in the body of this document. PMID- 11010671 TI - Health care programs: fraud and abuse; revised OIG civil money penalties resulting from public law 104-191. Office of Inspector General (OIG), HHS. Final rule. AB - This final rule revises the OIG's civil money penalty (CMP) authorities, in conjunction with new and revised provisions set forth in the Health Insurance Portability and Accountability Act of 1996. Among other provisions, this final rulemaking codifies new CMPs for excluded individuals retaining ownership or control interest in an entity; upcoding and claims for medically unnecessary services; offering inducements to beneficiaries; and false certification of eligibility for home health services. This rule also codifies a number of technical corrections to the regulations governing OIG's sanction authorities. PMID- 11010672 TI - Biofeedback in the treatment of urinary incontinence in adults. PMID- 11010673 TI - FDG positron emission tomography in head and neck cancer. PMID- 11010674 TI - High dose lymphoablative therapy (HDLT) with or without stem cell rescue for treatment of severe autoimmune diseases. PMID- 11010675 TI - Intradiscal electrothermal therapy for chronic low back pain. PMID- 11010676 TI - Pelvic floor electrical stimulation in the treatment of adult urinary incontinence. PMID- 11010678 TI - Cardiology. PMID- 11010677 TI - Intrathecal baclofen pump implantation. PMID- 11010679 TI - Standards for privacy of individually identifiable health information. Office of the Assistant Secretary for Planning and Evaluation, DHHS. Proposed rule. AB - This rule proposes standards to protect the privacy of individually identifiable health information maintained or transmitted in connection with certain administrative and financial transactions. The rules proposed below, which would apply to health plans, health care clearinghouses, and certain health care providers, propose standards with respect to the rights individuals who are the subject of this information should have, procedures for the exercise of those rights, and the authorized and required uses and disclosures of this information. The use of these standards would improve the efficiency and effectiveness of public and private health programs and health care services by providing enhanced protections for individually identifiable health information. These protections would begin to address growing public concerns that advances in electronic technology in the health care industry are resulting, or may result, in a substantial erosion of the privacy surrounding individually identifiable health information maintained by health care providers, health plans and their administrative contractors. This rule would implement the privacy requirements of the Administrative Simplification subtitle of the Health Insurance Portability and Accountability Act of 1996. PMID- 11010680 TI - No known cause: leukemia research. PMID- 11010681 TI - Medicare program; coverage of ambulance services and vehicle and staff requirements. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period revises and updates Medicare policy concerning ambulance services. It identifies destinations to which ambulance services are covered, establishes requirements for the vehicles and staff used to furnish ambulance services, and clarifies coverage of nonemergency ambulance services for Medicare beneficiaries. This rule also implements section 4531 (c) of the Balanced Budget Act of 1997 concerning Medicare coverage for paramedic intercept services in rural communities. PMID- 11010682 TI - Special Supplemental Nutrition Program for Women, Infants and Children (WIC): non discretionary funding provisions of the William F. Goodling Child Nutrition Reauthorization Act of 1998. Food and Nutrition Service, USDA. Final rule. AB - This final rule incorporates into the WIC program regulations numerous non discretionary funding provisions mandated in the William F. Goodling Child Nutrition Reauthorization Act of 1998. This rule revises and expands backspend and spendforward authority, conversion of funds, multipurpose/infrastructure grants and the use of food funds for the purchase of breast pumps. The rule also revises nutrition services and administration expenditure standards and expands the timing for the use of vendor and participant collections. The provisions in this rule provide greater flexibility for State agencies in the operation of WIC program relating to funds management. PMID- 11010683 TI - Chronic beryllium disease prevention program. Office of Environment, Safety and Health, Department of Energy. Final rule. AB - The Department of Energy (DOE) is today publishing a final rule to establish a chronic beryllium disease prevention program (CBDPP) to reduce the number of workers currently exposed to beryllium in the course of their work at DOE facilities managed by DOE or its contractors, minimize the levels of, and potential for, exposure to beryllium, and establish medical surveillance requirements to ensure early detection of the disease. This program improves and codifies provisions of a temporary CBDPP established by DOE directive in 1997. PMID- 11010684 TI - Special Supplemental Nutrition Program for Women, Infants, and Children (WIC): bloodwork requirements. Food and Nutrition Service, USDA. Final rule. AB - This final rule amends regulations governing the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) to allow State agencies the option to defer the collection of blood test data for up to 90 days after the date of certification, so long as the applicant is determined to have at least one qualifying nutrition risk factor at the time of certification. In addition, this final rule will expand the current regulatory standard of the maximum age of blood test data used to assess nutritional risk for WIC certification. Although blood tests may no longer be a mandatory part of each WIC applicant's certification intake process, such tests are still required for the purposes of assessing nutritional status, nutrition surveillance, providing nutrition education, further tailoring food packages to meet nutritional needs, and referring to appropriate health and social services in the community. PMID- 11010686 TI - Medicare program; solvency standards for provider-sponsored organizations. Health Care Financing Administration (HCFA), HHS. Final rule. AB - The Balanced Budget Act of 1997 established a new Medicare+Choice (M+C) program that offers eligible individuals Medicare benefits through enrollment in one of an array of private health plans that contract with us. Among the new options available to Medicare beneficiaries is enrollment in a provider-sponsored organization (PSO). This final rule revises and responds to comments on solvency standards that certain entities must meet to contract as PSOs under the new M+C program. These standards, originally established in an interim final rule published on May 7, 1998, apply to PSOs that have received a waiver of the requirement that M+C organizations must be licensed by a State as risk-bearing entities. PMID- 11010685 TI - Approval of Hospital/Medical/Infectious Waste Incinerator State Plan for designated facilities and pollutants: Indiana. Environmental Protection Agency (EPA). Direct final rule. AB - EPA is approving Indiana's State Plan for Hospital/Medical/Infectious Waste Incinerators (HMIWI), submitted on September 30, 1999. The State Plan adopts and implements the Emissions Guidelines (EG) applicable to existing HMIWIs. This approval means that EPA finds the State Plan meets Clean Air Act (Act) requirements. Once effective, this approval makes the State Plan federally enforceable. PMID- 11010687 TI - Tricare; Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); nonavailability statement requirement for maternity care. Office of the Secretary, DoD. Interim final rule. AB - This interim final rule implements Section 712(c) of the National Defense Authorization Act for Fiscal Year 2000 (Pub. L. No. 106-65), which requires that a nonavailability-of-health-care statement shall be required for a non-enrolled beneficiary for TRICARE cost-share of maternity care services related to outpatient prenatal, outpatient or inpatient delivery, and outpatient post-partum care subsequent to the visit which confirms the pregnancy. The Act reestablishes a requirement which was previously eliminated under the broad direction of The National Defense Authorization Act of FY 1997, section 734, which removed authority for nonavailability statements (NASs) for outpatient services. Therefore, the Act changes the existing provisions require an NAS for inpatient delivery but do not require an NAS for outpatient prenatal and post-partum care. The change will significantly contribute to continuity of care for maternity patients. In furtherance of that principle, and consistent with the previous policy, an NAS for maternity care shall not be required when a beneficiary has other health insurance for primary coverage. This is being issued as an interim final rule in order to comply with the statutory mandate. Public comments, however, are invited and will be considered in connection with possible revisions to this rule. PMID- 11010688 TI - Family and medical leave. Office of Personnel Management. Final rule. AB - The Office of Personnel Management is issuing final regulations on the Family and Medical Leave Act of 1993 to ensure that both employees' and agencies' rights are protected and their responsibilities fulfilled. PMID- 11010689 TI - Prescription Drug Marketing Act of 1987; Prescription Drug Amendments of 1992; policies, requirements, and administrative procedures; delay of effective date; reopening of administrative record. Food and Drug Administration, HHS. Final rule; delay of effective date; reopening of administrative record. AB - The Food and Drug Administration (FDA) is delaying until October 1, 2001, the effective date and reopening the administrative record to receive additional comments regarding certain requirements of a final rule published in the Federal Register of December 3, 1999 (64 FR 67720). The other provisions of the final rule become effective on December 4, 2000. The final rule implements the Prescription Drug Marketing Act of 1987 (PDMA), as modified by the Prescription Drug Amendments of 1992 (PDA) and the FDA Modernization Act of 1997 (the Modernization Act). FDA is delaying the effective date for certain requirements relating to wholesale distribution of prescription drugs by distributors that are not authorized distributors of record. FDA is also delaying the effective date of another requirement that would prohibit blood centers functioning as "health care entities" to act as wholesale distributors of blood derivatives. The agency is taking this action to address numerous concerns about the provisions raised by affected parties. PMID- 11010690 TI - Medical devices; revocation of cardiac pacemaker registry. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to revoke a regulation requiring a cardiac pacemaker registry. The registry, which was mandated by the Deficit Reduction Act of 1984, requires any physician and any provider of services who requests or receives Medicare payment for an implantation, removal, or replacement of permanent cardiac pacemaker devices and pacemaker leads to submit certain information to the registry. The information is used by FDA to track the performance of permanent cardiac pacemakers and pacemaker leads and by the Health Care Finance Administration (HCFA) to administer its Medicare payment program for these devices. This action is being taken to implement an act to Repeal An Unnecessary Medical Device Reporting Requirement passed by Congress in 1996 to remove the cardiac pacemaker registry to eliminate duplicative and unnecessary reporting. PMID- 11010691 TI - Progestational drug products for human use; requirements for labeling directed to the patient. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is revoking its regulation requiring patient labeling for progestational drug products. Patient labeling had been required to inform patients of an increased risk of birth defects reported to be associated with the use of these drugs during the first 4 months of pregnancy. FDA concluded that, based on a review of the scientific data, such labeling for all progestogens is not warranted. In addition, the diversity of drugs that can be described as progestational and the diversity of conditions these drugs may be used to treat make it inappropriate to consider these drugs a single class for labeling purposes. This action is intended to provide consumers with more appropriate labeling for certain drug products. PMID- 11010692 TI - Medicare program; suggestion program on methods to improve Medicare efficiency. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period establishes a program to encourage individuals to submit suggestions that could improve the efficiency of the Medicare program. The rule implements section 203(c) of the Health Insurance Portability and Accountability Act of 1996. The intent of this rule is to encourage suggestions and to award, if we deem appropriate, monetary payments to individuals for suggestions that improve efficiency and produce monetary savings to the Medicare program. PMID- 11010693 TI - Medicare program; revisions to payment policies under the physician fee schedule for calendar year 2000. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule makes several changes affecting Medicare Part B payment. The changes include: implementation of resource-based malpractice insurance relative value units (RVUs); refinement of resource-based practice expense RVUs; payment for physician pathology and independent laboratory services; discontinuous anesthesia time; diagnostic tests; prostate screening; use of CPT modifier -25; qualifications for nurse practitioners; an increase in the work RVUs for pediatric services; adjustments to the practice expense RVUs for physician interpretation of Pap smears; and revisions to the work RVUs for new and revised CPT codes for calendar year 1999 and a number of other changes relating to coding and payment. Furthermore, we are finalizing the 1999 interim physician work RVUs and are issuing interim RVUs for new and revised codes for 2000. This final rule solicits public comments on the second 5-year refinement of work RVUs for services furnished beginning January 1, 2002 and requests public comments on potentially misvalued work RVUs for all services in the CY 2000 physician fee schedule. This final rule also conforms the regulations to existing law and policy regarding: removal of the x-ray as a prerequisite for chiropractic manipulation; the exclusion of payment for assisted suicide; and optometrist services. This final rule also announces the calendar year 2000 Medicare physician fee schedule conversion factor under the Medicare Supplementary Insurance (Part B) program as required by section 1848(d) of the Social Security Act. The 2000 Medicare physician fee schedule conversion factor is $36.6137. PMID- 11010694 TI - Age, the stress process, and physical distress: the role of distal stressors. AB - OBJECTIVES: This article examines the nature of distal stressors over a 15-year period and the extent to which they contribute to the explanation of additional variation in the life stress model. METHODS: The study uses data collected from a community sample of adults to test specific hypotheses. RESULTS: Findings show that (a) distal stressors significantly increase variance explained in physical symptoms and maintain a direct effect on physical symptoms, (b) recent stressors mediate the effects of distal stressors on distress, and (c) social resources have direct and mediating effects on physical distress. Variations exist for different age groups or life stages. DISCUSSION: The implications of the findings are discussed in the context of life course, theoretical issues of temporality, types of stressors, and the role of social resources. The major conclusion is that there are multiple pathways to distress that differ for different stages in the life course. PMID- 11010695 TI - Factors that contribute to positive perceived health in an older population. AB - This study explored the extent to which factors commonly associated with negative outcomes of aging also predicted positive perceived health in a group of community-dwelling older people. Questionnaires originally administered during a randomized trial of HMO's outpatient group visit program supplied data from approximately 700 participants. Stepwise modeling identified significant factors within categories of predictors of perceived health. Hierarchical multiple linear regression then modeled the incremental importance of theoretically-derived categories of factors. Many but not all of the factors previously associated with negative outcomes of aging also predicted positive perceived health, accounting for 38% of the variation. The most important predictors were fewer chronic conditions and no worsening of those conditions, mobility and better physical performance status, and the absence of depression. Gender, limitations in daily living activities, dementia, and utilization of services had little effect. The categories of f factors associated with positive perceived health had cumulative, interdependent effects. PMID- 11010696 TI - Occurrence of hip fractures and socioeconomic position. AB - OBJECTIVES: The purpose of this study was to determine hip fracture incidence in the older U.S. White population as a function of their socioeconomic position. METHODS: A sample of 5,161 White, hip fracture cases, 50 years and older, was selected using data from the National Hospital Discharge Survey for 1989-91. Median annual household income by ZIP Code of residence based on the 1990 Census was used as the measure of socioeconomic position. Fracture rates were calculated by age, sex, and income groups ranging from under $20,000 to $40,000 and more. RESULTS: A weighted, least squares analysis found a significant linear decrease in rates with increasing income after controlling for age and sex. CONCLUSIONS: The results indicate that hip fracture incidence varies as a function of the income level of the ZIP Code area where the population resides. Implications for targeting prevention programs within local areas with low median income are discussed. PMID- 11010697 TI - A health and demographic profile of noninstitutionalized older Americans residing in environments with home modifications. AB - OBJECTIVES: In this analysis the authors investigate the demographic characteristics, the health conditions/events, and the disabilities of community dwelling Americans 70 years of age and older that are associated with residing in environments with specific home modifications. METHODS: Data from a large population-based study of the elderly are used to estimate logistic regression equations that reveal profiles of older individuals who are likely to have distinct home modifications. RESULTS: Having diseases such as diabetes and stroke, having experienced a hip fracture, a fall or a joint replacement, and having greater limitations with activities of daily living raise the likelihood of having home modifications. Low income, Hispanic, and African American elderly appear underrepresented among those with modifications. DISCUSSION: The authors conclude that specialized housing alternatives will be an increasingly important issue in the future as individuals aim to achieve and maintain the delicate balance between their functional ability and their living environment. PMID- 11010698 TI - Race and ethnic variation in the disablement process. AB - OBJECTIVES: This analysis examines ethnoracial group differences in the transition from health to disability. METHODS: Using data from the AHEAD study, the authors examine the relative influence of each stage in the disablement process in the evolution of ethnoracial group differences in basic and instrumental disability. RESULTS: Predisposing factors account for disability differences between Whites and other Latinos, whereas excess disability among African Americans stems from their higher level of cognitive limitation. The excess disability of Mexican Americans arises from their higher level of physical limitations. The data also reveal a larger impact of medical conditions and physical limitations on acquisition of disability among African Americans and Mexican Americans. This article demonstrates the importance of cognitive status in the disablement process, especially in ethnoracial group differences. DISCUSSION: The authors discuss the practical implications for health care delivery to non-White elders and the theoretical implications for understanding the complexities of disablement. PMID- 11010699 TI - Outcomes of a small group educational intervention for urinary incontinence: episodes of incontinence and other urinary symptoms. AB - OBJECTIVES: This community-based intervention to reduce urinary incontinence (UI) in elderly women used a small group educational approach. This article reports on change in episodes of incontinence and other urinary symptoms. METHODS: Participants were randomly assigned to intervention or wait control condition. This article is restricted to 49 intervention and 59 control participants with acceptable diaries. RESULTS: There was a significant treatment effect for a number of incontinent episodes. In the intervention group, 61% had a 50% or greater reduction in episodes, with more than one third having 100% reduction; 38% of the control group had a reduction of 50% or greater. One year postprogram, 75% of treated women reported subjective improvement. There was a reduction in frequency of daily, but not nocturnal, micturition. DISCUSSION: This community based intervention is an encouraging option for behavioral treatment of UI. Public health models may be particularly appropriate with moderate levels of urinary incontinence. PMID- 11010700 TI - Must poor kids have bad teeth? PMID- 11010701 TI - General and plastic surgery devices; classification of the nonresorbable gauze/sponge for external use, the hydrophilic wound dressing, the occlusive wound dressing, and the hydrogel wound dressing. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the nonresorbable gauze/sponge for external use, the hydrophilic wound dressing, the occlusive wound dressing, and the hydrogel wound dressing into class I (general controls). FDA is also exempting these devices from premarket notification procedures. This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976 (the 1976 amendments), the Safe Medical Devices Act of 1990 (SMDA), and the Food and Drug Administration Modernization Act of 1997 (FDAMA). PMID- 11010702 TI - Human drugs and biologics; determination that informed consent is NOT feasible or is contrary to the best interests of recipients; revocation of 1990 interim final rule; establishment of new interim final rule. Food and Drug Administration, HHS. Interim final rule; opportunity for public comment. AB - The Food and Drug Administration (FDA) is revoking its 1990 interim final regulations that permitted the Commissioner of Food and Drugs (the Commissioner) to determine that obtaining informed consent from military personnel for the use of an investigational drug or biologic is not feasible in certain situations related to military combat. FDA also is issuing a new interim final rule addressing waiver of informed consent in military operations. FDA is taking these actions based on its analysis and consideration of all relevant facts, including its evaluation of the Department of Defense's (DOD) experience during the Persian Gulf War, its evaluation of the comments received by the agency in response to the agency's July 31, 1997, request for comments on whether the agency should revise or revoke the interim regulations, and the enactment of the Strom Thurmond National Defense Authorization Act for Fiscal Year 1999 (the Defense Authorization Act). Under the Defense Authorization Act, the President is authorized to waive the Federal Food, Drug, and Cosmetic Act's (the act) informed consent requirements in military operations if the President finds that obtaining consent is infeasible or contrary to the best interests of recipients and on an additional ground that obtaining consent is contrary to national security interests. In light of the enactment of the Defense Authorization Act, with an immediate effective date, and because the President could be called upon to make a waiver determination for military personnel engaged in a specific military operation at any time, the agency believes that it is critical to have in place adequate criteria and standards for the President to apply in making an informed consent waiver determination. Therefore, FDA is issuing a new interim final regulation with an immediate effective date to establish criteria and standards for the President to apply in making a determination that informed consent is not feasible or is contrary to the best interests of the individual recipients. PMID- 11010703 TI - Organ Procurement and Transplantation Network. Health Resources and Services Administration, HHS. Final rule. AB - This document sets forth improvements to the final rule governing the operation of the Organ Procurement and Transplantation Network (OPTN), published in 1998. It reflects the advice of a panel convened by the National Academy of Science's Institute of Medicine, as called for in the Department's appropriation act for 1999. It also reflects comments on the 1998 rule and consultation with representatives of the organ transplantation community, as recommended in the same legislation; and it summarizes new transplant data developed in the period since enactment of the appropriations act. PMID- 11010704 TI - Enrollment--provision of hospital and outpatient care to veterans. Department of Veterans Affairs. Final rule. AB - This document amends VA's medical regulations. The Veterans' Health Care Eligibility Reform Act of 1996 mandates that VA implement a national enrollment system to manage the delivery of healthcare services. Accordingly, the medical regulations are amended to establish provisions consistent with this mandate. Starting October 1, 1998, most veterans were required to be enrolled in the VA healthcare system as a condition of receiving VA hospital and outpatient care. Veterans will be allowed to apply to be enrolled at any time. They will be eligible to be enrolled based on funding availability and their priority status. In accordance with statutory provisions, the final rule also states that some categories of veterans are eligible for VA hospital and outpatient care even if not enrolled. This document further establishes a "medical benefits package" setting forth, with certain exceptions, the hospital and outpatient care that will be provided to enrolled veterans and certain other veterans. Moreover, this document announces that VA will enroll all 7 priority categories of veterans for the period October 1, 1999 through September 30, 2000, unless it is necessary to change this determination by a subsequent rulemaking document. PMID- 11010705 TI - Currently effective Indian Health Service eligibility regulations. Indian Health Service, HHS. Republication of currently effective Indian Health Service eligibility regulations. AB - The HHS is publishing in the Federal Register, final regulations governing eligibility for services from the Indian Health Service. The eligibility regulations currently codified at 42 CFR part 36 are under a congressional moratorium. Republishing the regulations that are currently in effect while the codified regulations are under moratorium is being done for the convenience of the public and in conformance with the requirement of the Administrative Procedure Act, 5 U.S.C. 552(a)(1), that the Code of Federal Regulations (CFR) must contain currently effective regulations. PMID- 11010706 TI - Food labeling: health claims; soy protein and coronary heart disease. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is authorizing the use, on food labels and in food labeling, of health claims on the association between soy protein and reduced risk of coronary heart disease (CHD). Based on its review of evidence submitted with comments to the proposed rule, as well as evidence described in the proposed rule, the agency has concluded that soy protein included in a diet low in saturated fat and cholesterol may reduce the risk of CHD by lowering blood cholesterol levels. PMID- 11010708 TI - Medicare program; revision to accrual basis of accounting policy. Health Care Financing Administration (HCFA), HHS. Final rule. AB - Medicare policy provides that payroll taxes that a provider becomes obligated to remit to governmental agencies are included in allowable costs only in the cost reporting period in which payment (upon which the payroll taxes are based) is actually made to an employee. Therefore, for payroll accrued in 1 year but not paid until the next year, the associated payroll taxes are not an allowable cost until the next year. This final rule provides for an exception when payment would be made to the employee in the current year but for the fact the regularly scheduled payment date is after the end of the year. In that case, the rule requires allowance in the current year of accrued taxes on payroll that is accrued through the end of the year but not paid until the beginning of the next year, thus allowing accrued taxes on end-of-the year payroll in the same year that the accrual of the payroll itself is allowed. The effect of this rule is not on the allowability of cost but rather only on the timing of payment; that is, the cost of payroll taxes on end-of-the-year payroll is allowable in the current period rather than in the following period. PMID- 11010709 TI - Medical devices; gastroenterology and urology devices; classification of the electrogastrography system. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the electrogastrography system (EGG) into class II (special controls). The special controls that will apply to the EGG system are restriction to prescription use, certain labeling requirements, design requirements, and data collection requirements. The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997. The agency is classifying the EGG system into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. PMID- 11010710 TI - Health standards for occupational noise exposure. Mine Safety and Health Administration (MSHA), Labor. Final rule. AB - This final comprehensive rule replaces MSHA's existing standards for occupational noise exposure in coal mines and metal and nonmetal mines. The final rule establishes uniform requirements to protect the Nation's miners from occupational noise-induced hearing loss. The rule is derived in part from existing MSHA noise standards, and from the Department of Labor's existing occupational noise exposure standard for general industry promulgated by the Occupational Safety and Health Administration (OSHA). As a result of the Agency's ongoing review of its safety and health standards, MSHA determined that its existing noise standards, which are more than twenty years old, do not adequately protect miners from occupational noise-induced hearing loss. A significant risk to miners of material impairment of health from workplace exposure to noise over a working lifetime exists when miners' exposure exceeds an 8-hour time-weighted average (TWA8) of 85 dBA. MSHA expects that the final rule will significantly reduce the risk of material impairment within the mining industry as a whole. PMID- 11010711 TI - Recent trends in children's health insurance coverage: no gains for low-income children. AB - The percentage of low-income children who have health insurance has not changed over the last few years, despite expansions in public coverage through Medicaid and the State Children's Health Insurance Program (SCHIP). Data from 1996-1997 and 1998-1999 from the Center for Studying Health System Change (HSC) find that while the proportion of low-income children with public coverage has increased, the percentage with private insurance coverage has decreased sharply, resulting in no net change in the percentage who are uninsured. This Issue Brief describes these recent changes in public and private coverage. Possible factors that may explain these changes are discussed, including increases in private insurance premiums, substitution of public for private coverage and changes in the characteristics of low-income persons. The study did not determine conclusively the causes of the changes in coverage. PMID- 11010712 TI - Waging a war on drugs: administering a lethal dose to Kendra's law. PMID- 11010713 TI - State child health; State Children's Health Insurance Program allotments and payments to states. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This rule sets forth the methodologies and procedures to determine the allotments of Federal funds for each Federal fiscal year (FY) available to individual States, Commonwealths and Territories under title XXI of the Social Security Act. This rule also specifies the allotment, payment, and grant award process that will be used for the States, the Commonwealths and Territories to claim and receive Federal financial participation (FFP) for expenditures under the State Children's Health Insurance Program (SCHIP) and related Medicaid program provisions. Established by section 4901 of the Balanced Budget Act of 1997 (Public Law 105-33), amended by technical amendments (made by Public Law 105 100), and most recently amended by the Medicare, Medicaid and SCHIP Balanced Budget Refinement Act (BBRA) of 1999 (Public Law 106-113, enacted November 29, 1999), the State Children's Health Insurance Program provides Federal matching funds to States to initiate and expand health insurance coverage to uninsured, low-income children. Aggregate Federal funding is limited to a fixed amount for each Federal fiscal year. This aggregate amount is divided into allotments for each State. State allotments are determined based on a statutory formula that divides the total available appropriation among all States with approved child health plans. Once determined, the amount of a State's allotment for a fiscal year is available for 3 years. We are publishing this final rule in accordance with the provisions of sections 2104 and 2105 of the Act that relate to allotments and payments to States under title XXI. PMID- 11010714 TI - Addition of medical criteria for evaluating Down syndrome in adults. Social Security Administration (SSA). Final rule. AB - We are adding a new listing to evaluate non-mosaic Down syndrome in adults. Our current regulations only include a listing for evaluating Down syndrome in children; we evaluate claims filed by adults with Down syndrome under other listings. We are establishing a separate adult listing for this disorder to acknowledge its lifelong impact and severity. We expect that these final rules will simplify and expedite our adjudication of claims filed by adults with non mosaic Down syndrome. PMID- 11010715 TI - Obstetrical and gynecological devices; classification of female condoms. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is classifying the preamendments female condom intended for contraceptive and prophylactic purposes. Under this rule, the preamendments female condom is being classified into class III (premarket approval). This action is being taken under the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the FDA Modernization Act of 1997. PMID- 11010716 TI - Hematology. PMID- 11010717 TI - Radial artery harvesting. PMID- 11010718 TI - Pharmaceutical marketplace dynamics. PMID- 11010719 TI - Sterility requirement for aqueous-based drug products for oral inhalation. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending its regulations to require that all prescription and over-the-counter (OTC) aqueous-based drug products for oral inhalation be manufactured sterile. This rule applies to aqueous-based oral inhalation drug products in both single-dose and multiple-use primary packaging. Pressurized metered-dose inhalers are not subject to this rule. Based on reports of adverse drug experiences from contaminated nonsterile inhalation drug products and recalls of these products, FDA is taking this action to help ensure the safety and effectiveness of these products. PMID- 11010720 TI - Nondiscrimination on the basis of disability in air travel; compensation for damage to wheelchairs and other assistive devices. Office of the Secretary, DOT. Final rule. AB - The Department is amending its rules implementing the Air Carrier Access Act of 1986 (ACAA) to lift an existing cap on the amount of compensation airlines have to pay to passengers for loss or damage of their wheelchairs and other assistive devices. The rule is intended to provide additional relief to passengers who use expensive assistive devices that are lost, destroyed or damaged in the course of airline travel. PMID- 11010721 TI - Federal Employees Health Benefits (FEHB) Program and Department of Defense (DoD) demonstration project. Office of Personnel Management. Final rule. AB - OPM is issuing a final regulation to implement the portion of the National Defense Authorization Act for 1999 that establishes authority for a demonstration project under which certain Medicare and other eligible DoD beneficiaries can enroll in health benefit plans in certain geographic areas under the Federal Employees Health Benefits (FEHB) Program. The demonstration project will run for a period of three years from January 1, 2000, through December 31, 2002. This regulation specifies only the requirements that differ from existing FEHB Program regulations because of unique aspects of the demonstration project. PMID- 11010722 TI - Children suffering from spina bifida who are children of Vietnam veterans. Department of Veterans Affairs. Final rule. AB - This document amends Department of Veterans Affairs (VA) regulations concerning benefits for children suffering from spina bifida who are children of Vietnam veterans. This is necessary to implement statutory changes contained in the Veterans' Benefits Act of 1997. PMID- 11010723 TI - The State Vocational Rehabilitation Services Program. Office of Special Education and Rehabilitative Services, Department of Education. Final regulations. AB - The Secretary amends the regulations governing The State Vocational Rehabilitation Services Program (VR program). These amendments are needed to implement changes in the Rehabilitation Act of 1973, as amended (Act). These changes establish evaluation standards and performance indicators for the VR program. PMID- 11010724 TI - Privacy Act; implementation. Office of Inspector General (OIG), HHS. Final rule. AB - This final rule exempts the new system of records, the Healthcare Integrity and Protection Data Bank (HIPDB), from certain provisions of the Privacy Act (5 U.S.C. 552a). The establishment of the HIPDB is required by section 1128E of the Social Security Act (the Act), as added by section 221(a) of the Health Insurance Portability and Accountability Act (HIPAA) of 1996. Section 1128E of the Act directed the Secretary to establish a national health care fraud and abuse data collection program for the reporting and disclosing of certain final adverse actions taken against health care providers, suppliers or practitioners, and to maintain a data base of final adverse actions taken against health care providers, suppliers and practitioners. Regulations implementing the new HIPDB were published in the Federal Register on October 26, 1999 (64 FR 57740). The exemption being set forth in this rule applies to investigative materials compiled for law enforcement purposes. PMID- 11010725 TI - Medicare program; State Health Insurance Assistance Program (SHIP). Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This interim final rule explains the terms and conditions that apply to grants to States for counseling and assistance to Medicare beneficiaries, and makes several minor technical clarifications about program compliance. We also specify our policies regarding the treatment of funds associated with the management of this program, including user fee assessments not in effect when prior regulations were issued. This interim final rule is issued in accordance with section 4360 of the Omnibus Budget Reconciliation Act of 1990 (OBRA '90) and section 1857(e)(2) of the Social Security Act (the Act). PMID- 11010726 TI - Investigational new drug applications; amendment to clinical hold regulations for products intended for life-threatening diseases and conditions. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending the regulations governing investigational new drug applications (IND's) to permit FDA to place a clinical hold on one or more studies under an IND involving a drug that is intended to treat a life-threatening disease or condition affecting both genders. The amendments permit the agency to place a clinical hold on such studies if men or women with reproductive potential who have the disease or condition are otherwise eligible but are categorically excluded from participation solely because of a perceived risk or potential risk of reproductive or developmental toxicity from use of the investigational drug. This rule was developed in response to the past practice of excluding women with reproductive potential from early clinical trials because of a perceived risk or potential risk of reproductive or developmental toxicity. The final rule does not impose requirements to enroll or recruit a specific number of men or women with reproductive potential. PMID- 11010727 TI - Federal old-age, survivors and disability insurance and Supplemental Security Income for the Aged, Blind, and Disabled; medical and other evidence of your impairment(s) and definition of medical consultant. Social Security Administration. Final rules. AB - We are revising the Social Security and Supplemental Security Income (SSI) disability regulations regarding sources of evidence for establishing the existence of a medically determinable impairment under title II and title XVI of the Social Security Act (the Act). We are doing this to clarify and expand the list of acceptable medical sources and to revise the definition of the term "medical consultant" to include additional acceptable medical sources. PMID- 11010728 TI - Managed long-term care: what role do Area Agencies on Aging play? PMID- 11010729 TI - Beyond 43 million: the "regarded as" prong of the ADA and HIV infection--a tautological approach. PMID- 11010730 TI - Effects of a program of intervention on parental distress following infant death. AB - A longitudinal study of 144 parents (65 fathers, 79 mothers) was conducted to evaluate the effectiveness of a program of intervention in relieving the psychological distress of parents affected by infant death. Participants were assessed in terms of their psychiatric disturbance, depression, anxiety, physical symptoms, dyadic adjustment, and coping strategies. The experimental group (n = 84) was offered an intervention program comprising the use of specially designed resources and contact with a trained grief worker. A control group (n = 60) was given routine community care. Parental reactions were assessed at four to six weeks postloss (prior to the implementation of the intervention program), at six months postloss, and at 15 months postloss. A series of multivariate analyses of variance revealed that the intervention was effective in reducing the distress of parents, particularly those assessed prior to the intervention as being at high risk of developing mourning difficulties. Effects of the intervention were noted in terms of parents' overall psychiatric disturbance, marital quality, and paternal coping strategies. PMID- 11010731 TI - The do-not-resuscitate decision: the context, process, and consequences of DNR orders. AB - This study examines the process and consequences of an increasingly important element of the dying experience in American hospitals: the writing of a Do-Not Resuscitate (DNR) order. The focus of the study is on the decision-making process and timing of the DNR decision, the impact of the DNR order on the dying experience, and the consequences of the DNR order for length of hospital stay and accrued medical charges. Patients with a DNR order are compared to those who were unsuccessfully coded. Data are obtained from a review and analysis of the medical charts and death monitor sheets of a sample of 249 persons who died in 1994 in a single teaching hospital. The study found physicians routinely discuss the DNR decision with patients and/or their surrogates (though patients are involved in the decision in only about one-third of cases) and that the decision is often made relatively early in the hospital stay. The dying experience of patients with a written DNR was different in significant ways from the experience of unsuccessfully-coded patients. Those with a DNR were more likely to remain in a single unit in the hospital and less likely to die in an intensive care unit or while connected to a ventilator. Consistent with other studies, however, average length of hospital stay and average medical charges were actually higher for the DNR patients. Implications of these differences between DNR and unsuccessfully coded patients are discussed. PMID- 11010732 TI - Death education: knowledge, attitudes, and perspectives of Irish parents and teachers. AB - This article reports on a cross-sectional survey of the knowledge, attitudes and perspectives of Irish parents and school teachers concerning children's grief and the concept of death education. The sample comprised 119 parents and 142 teachers of Irish Primary-school children (5-12 years of age) who completed a self administered questionnaire. Both parents and teachers reported high levels of understanding of the nature of children's grief and strongly supported the view that death should be discussed with children before they encounter it. Although discussions of death were reported in the classroom and in the home, both teachers and parents, particularly men, reported being uncomfortable talking to children about death. There was general support for inclusion of death education in the school curriculum, with both teachers and parents supporting the need for further teacher training to undertake its delivery. There were few significant differences between the expressed attitudes of parents and teachers. However, teachers were more likely than parents to agree that death education would take away from parental responsibility. The implications of the findings for further work in this area are considered. PMID- 11010733 TI - Death experiences and hospice: perceptions of college students. AB - The purpose of this study was to investigate college-age students' awareness of and involvement in hospice and to ascertain information regarding their own experiences with death and their preferences about death and dying. Our data were gathered through a mailed survey to 521 biology majors at an undergraduate college in the southeastern United States (122 responded or 23%). The majority of respondents were aware of hospice, yet less than 25% had any involvement with hospice. Over 90% of the respondents had seen a dead body, and one-third had witnessed a death. Statistically significant differences by gender were found only with awareness of hospice, with women being more knowledgeable. With health professions as the goal for most of these students, a similar study with a cohort of non-college students, or even non-health-profession-bound college students, would serve as an interesting "control" group. PMID- 11010734 TI - Federal Employees Health Benefits (FEHB) program and Department of Defense (DoD) demonstration project; and other miscellaneous changes. Office of Personnel Management. Final rule. AB - OPM is issuing a final regulation to implement the portion of the Defense Authorization Act for 1999 that establishes authority for a demonstration project under which certain Medicare and other eligible DoD beneficiaries can enroll in health benefit plans in certain geographic areas under the Federal Employees Health Benefits (FEHB) Program. The demonstration project will run for a period of three years from January 1, 2000, through December 31, 2002. This regulation specifies only the requirements that differ from existing FEHB Program regulations because of unique aspects of the demonstration project. This regulation also makes other miscellaneous changes to the Federal Employees Health Benefits Acquisition Regulations. PMID- 11010735 TI - Privacy Act of 1974; proposed implementation. Office of the Secretary, HHS. Final rule. AB - The Department of Health and Human Services is exempting a new system of records, 09-25-0213, "Administration: Investigative Records, HHS/NIH/OM/OA/OMA," from certain requirements of the Privacy Act to protect records compiled in the course of an inquiry and/or investigation and to protect the identity of confidential sources who furnish information to the Government under an express promise that the identity of such source would be held in confidence. PMID- 11010736 TI - Sick leave for family care purposes. Office of Personnel Management. Final rule. AB - The Office of Personnel Management is issuing final regulations to expand the use of sick leave for family care purposes. Under the final regulations, an employee may use a total of up to 12 weeks of accrued sick leave each year to care for a family member with a serious health condition. This benefit broadens the options available for employees to meet their family responsibilities. PMID- 11010737 TI - HCFA's outpatient PPS: finally ready to roll? PMID- 11010738 TI - Name brands: the effects of intrusive HIV legislation on high-risk demographic groups. PMID- 11010739 TI - Confusion in the courts: managed care financial structures and their impact on medical care. PMID- 11010740 TI - Abacavir: new preparation. Risks limit the value. AB - (1) Abacavir, a nucleoside inhibitor of HIV reverse transcriptase, is indicated for the treatment of HIV-infected adults, in combination with other antiretroviral drugs. (2) The assessment file is limited mainly to trials involving patients who have never received other antiretroviral drugs. There are no comparisons of abacavir plus two other nucleoside inhibitors versus three nucleoside inhibitors or versus one non nucleoside inhibitor plus two nucleoside inhibitors. (3) According to initial results the optimal dose regimen of abacavir in adults appears to be 300 mg twice a day. The lamivudine + zidovudine + abacavir combination reduces viral load more strongly than the lamivudine + zidovudine combination in patients not previously treated with antiretroviral drugs. Preliminary results suggest that the same applies to patients previously treated with antiretroviral drugs. (4) A trial involving 562 previously untreated patients showed that the effect of the abacavir + lamivudine + zidovudine combination (total of 4 tablets a day) was not significantly different from that of the indinavir + lamivudine + zidovudine combination (8 tablets a day) after 48 weeks of treatment. (5) In clinical trials, approximately 3% of adults and 5% of children had hypersensitivity reactions to abacavir. Under the strict conditions of the trials it was not always possible to avoid abacavir rechallenge, which carries a risk of a new, potentially fatal hypersensitivity reaction. (6) The risk-benefit ratio of abacavir is currently unfavourable in children, who must not be prescribed the drug. (7) The risk of drug interactions with abacavir is low. PMID- 11010741 TI - Alendronic acid in primary prevention: new indication. No reduction in fracture risk. AB - (1) Alendronic acid at a dose of 5 mg/day is now licensed in France for primary prevention of postmenopausal fractures. (2) The clinical file is relatively bulky and methodologically adequate, but there are no comparisons with combined hormone replacement therapy or with raloxifen. (3) Three trials have shown that 5 mg/day alendronic acid slows postmenopausal bone loss. However, this effect disappears on treatment cessation, and mineral bone density is only one risk factor for postmenopausal fractures. (4) A placebo-controlled trial of primary prevention involving more than 4,000 patients showed no reduction in the risk of fracture after 4 years of treatment with alendronic acid (5 mg/day for 2 years, then 10 mg/day). (5) Alendronic acid increases the risk of oesophageal ulceration, necessitating strict precautions during ingestion. PMID- 11010742 TI - Ribavirin: new preparation. An advance, but still no cure-all. AB - (1) Ribavirin is licensed for patients with hepatitis C who have not been treated previously or who have relapsed. According to the licence, ribavirin must be combined with interferon alfa-2b. (2) The clinical file is bulky and of acceptable methodological quality. (3) Some trials have shown that ribavirin monotherapy has no effect on viral load. Two trials involving patients who had relapsed after transaminase normalisation on interferon alfa-2b monotherapy show that the ribavirin + interferon alfa-2b combination is more effective than interferon alfa-2b alone in rendering viraemia undetectable and normalising transaminase activity for at least 6 months after treatment cessation. (4) A trial of patients resistant to interferon alfa monotherapy showed that it was more beneficial to combine ribavirin + interferon alfa-2b than to double the dose of interferon alfa-2b alone, in terms of driving viraemia below the detection limit and persistently normalising transaminase activity. (5) Three trials involving patients who had not been treated previously showed that the ribavirin + interferon alfa-2b combination was more effective than interferon alfa-2b monotherapy in driving viraemia below the detection limit for at least 6 months after treatment cessation, and in reducing the liver inflammation score. No effect on fibrosis or on the risk of cirrhosis has yet been demonstrated. (6) The best duration of treatment is not known, but seems to depend on prognostic factors. The following factors might permit shorter treatment: relatively low viral load, early-stage fibrosis, female sex, age below 40 years, and infection by genotype 2 or 3 strains. (7) The ribavirin + interferon alfa-2b combination causes haemolytic anaemia in most patients, and this can be severe. (8) Ribavirin is teratogenic and causes sperm defects. PMID- 11010743 TI - Goserelin and locally advanced prostate cancer: new indication. Pros and cons. AB - (1) Goserelin, a GnRH agonist, has a new licensed indication in France, as an adjuvant to external radiotherapy for locally advanced prosate cancer. (2) The clinical file in this indication includes two trials of satisfactory methodological quality comparing radiotherapy + goserelin with radiotherapy alone. (3) In these trials the radiotherapy + goserelin combination increased the specific-symptom-free survival time. (4) In one trial goserelin caused endocrine disorders in 19% of patients. There were also more cases of urinary incontinence (13% in absolute values) among patients receiving the radiotherapy + goserelin combination. Furthermore, goserelin almost always causes impotence and reduced libido. PMID- 11010744 TI - Lidocaine + prilocaine before 3 months of age: new indication. Correct use is crucial. AB - (1) The lidocaine + prilocaine combination, as a cream or adhesive dressings, is now approved for anaesthesia of healthy skin in children of 0 to 3 months, an age range in which it used to be contraindicated. (2) The clinical file on the lidocaine + prilocaine combination in this age range is of acceptable methodological quality. (3) For circumcision, two trials showed that the lidocaine + prilocaine cream was more effective than a placebo cream in preventing pain. Two other trials showed that the lidocaine + prilocaine cream was less effective than penile block or local anaesthetic injection in newborns. (4) During heel blood sampling in newborns, the two available comparative trials showed no analgesic efficacy of the lidocaine + prilocaine combination. However, heel blood sampling is an unnecessary procedure. (5) The two comparative trials involving venepuncture in newborns showed, at best, a moderate analgesic effect of the lidocaine + prilocaine combination. (6) Cases of symptomatic methaemoglobinaemia have occurred when recommended conditions of use are not observed. PMID- 11010745 TI - Dexamethasone misused for weight gain. PMID- 11010746 TI - Bronchospasm with zanamivir. PMID- 11010747 TI - Antiplatelet drugs in cardiovascular prevention: introduction. PMID- 11010748 TI - Antiplatelet drugs in cardiovascular prevention: take adverse effects and costs into account. AB - (1) Several antiplatelet drugs have proven preventive efficacy, including aspirin, aspirin + dipyridamole, clopidogrel, ticlopidine and flurbiprofen. They differ mainly in their adverse effects and costs. (2) Aspirin has essentially gastrointestinal adverse effects, whose incidence can be limited by prescribing a daily dose below 350 mg. (3) The addition of dipyridamole to aspirin can cause headache and diarrhoea. (4) Ticlopidine should be avoided because of the risk of agranulocytosis. (5) Adverse effects are less frequent and less severe with clopidogrel than with ticlopidine. (6) Like other nonsteroidal antiinflammatory drugs, flurbiprofen has mainly gastrointestinal adverse effects. (7) Treatment is least costly with low-dose aspirin and most costly with clopidogrel. PMID- 11010749 TI - Antiplatelet drugs in cardiovascular prevention: coronary events: acute phase and secondary prevention. AB - (1) Aspirin reduces acute-phase mortality after myocardial infarction, and also reduces the risk of myocardial infarction and death in patients with unstable angina. Aspirin reduces the risk of myocardial infarction in patients with stable angina and after unstable angina, and the risk of relapse after myocardial infarction. It reduces the risk of complications during coronary angioplasty, and the risk of venous coronary bypass graft occlusion after coronary surgery. (2) The best risk-benefit ratio with aspirin is achieved at a daily dose of 75-350 mg; 160 mg/day is the best-validated dose in the acute phase of myocardial infarction. (3) Aspirin must be combined with a thrombolytic agent in patients with myocardial infarction, and with heparin in patients with unstable angina. During coronary stenting the aspirin + clopidogrel combination may have a better risk-benefit ratio than the aspirin + ticlopidine combination. (4) Clopidogrel can be used when aspirin is contraindicated or poorly tolerated. (5) Oral anticoagulants seem a better option than aspirin after complicated myocardial infarction. PMID- 11010750 TI - Antiplatelet drugs in cardiovascular prevention: stroke: acute phase and secondary prevention. AB - (1) In the acute phase of ischaemic stroke in patients free of thrombogenic heart disease, combined treatment with aspirin + moderate-dose unfractionated heparin reduces the risk of relapse and death. Unfractionated heparin at higher anticoagulant doses has an unfavourable risk-benefit ratio. Treatment is controversial in patients with events associated with atrial fibrillation. (2) After ischaemic stroke in patients free of thrombogenic heart disease, aspirin reduces the risk of relapse and death. Other antiplatelet drugs, the aspirin + dipyridamole combination, ticlopidine and clopidogrel have similar efficacy to aspirin. (3) The risk-benefit ratio of oral anticoagulant is favourable after ischaemic stroke associated with atrial fibrillation; but it is unfavourable after stroke without thrombogenic heart disease. PMID- 11010751 TI - Antiplatelet drugs in cardiovascular prevention: stroke prevention in patients with thrombogenic heart disease. AB - (1) In patients with atrial fibrillation and a moderate embolic risk, aspirin reduces the risk of stroke and has a comparable risk-benefit ratio to oral anticoagulants. (2) Oral anticoagulants are superior to aspirin in patients with atrial fibrillation and a history of stroke. (3) In patients with a mechanical valve prosthesis and a high embolic risk, the oral anticoagulant + aspirin combination has a better risk-benefit ratio than oral anticoagulant alone. PMID- 11010752 TI - Antiplatelet drugs in cardiovascular prevention: coronary events and stroke: primary prevention. AB - (1) Aspirin reduces the risk of myocardial infarction in men over 40 with no history of cardiovascular disease, and in hypertensive patients of both sexes over that age. But it does not seem to reduce overall mortality, and its risk benefit ratio is not very favourable because of its gastrointestinal adverse effects. (2) In patients with symptomatic lower-limb arterial disease, ticlopidine and clopidogrel reduce the risk of coronary events. PMID- 11010753 TI - Prevention of mother-child HIV transmission: progress and problems. AB - (1) Since 1994, zidovudine administration to pregnant women and their newborns has been the reference prophylaxis for mother-child HIV transmission. (2) Three trials done in Thailand and Africa show that zidovudine therapy during the last few weeks of pregnancy reduces the risk of transmission, especially when breast feeding is avoided. (3) A retrospective study done in the United States suggests that even very late zidovudine administration may benefit the mother and/or her child. (4) A trial done in Uganda suggests that a single dose of nevirapine has better preventive activity than a very short course of zidovudine in women who breastfeed. (5) There are very few data on the risk-benefit ratio of multidrug antiretroviral regimens in pregnant women. (6) Eight cases of mitochondrial involvement have been diagnosed in France in children exposed to zidovudine or to the zidovudine + lamivudine combination. These cases call for long-term close monitoring of children exposed to antiretroviral drugs in utero. To limit the duration of fetal exposure, the third trimester of pregnancy is now recommended as the safest time to start antiretroviral prophylaxis. (7) Despite a randomised trial and a meta-analysis suggesting that elective caesarean section prevents mother-child HIV transmission, the value of such surgery remains controversial. PMID- 11010754 TI - Delicate operations. PMID- 11010755 TI - Out-of-pocket spending on health care by Medicare beneficiaries age 65 and older: 1999 projections. PMID- 11010756 TI - Medicare beneficiaries and prescription drug coverage: gaps and barriers. PMID- 11010757 TI - Care management: policy considerations for original Medicare. PMID- 11010758 TI - Social Security reform: economic effects of planned changes in the CPI. PMID- 11010759 TI - Medical error and patient injury: costly and often preventable. PMID- 11010760 TI - Social Security reform: how do minorities fare under Social Security? A response to two Heritage Foundation reports. PMID- 11010761 TI - Social Security reform: the Thrift Savings Plan (TSP) for federal employees. PMID- 11010762 TI - Determining comparable levels of functional disability. PMID- 11010763 TI - A critical look at equity investment in the 1994-1996 Advisory Council on Social Security recommendations. PMID- 11010764 TI - Consumer-directed long-term care: participants' experiences in five countries. PMID- 11010765 TI - Social Security reform: rethinking retirement-age policy--a look at raising Social Security's retirement age. PMID- 11010766 TI - Fractures of the mandibular condyle. Part 1: patterns of distribution of types and causes of fractures in 348 patients. AB - This prospective study was designed to record relevant characteristics of mandibular condyle fractures and to evaluate the relationship between these. Data were recorded on sex, age, cause of trauma, level of fracture, dislocation of the mandibular head, dental state and associated fractures of all patients diagnosed in our hospital during the period 1984-1996 with mandibular condyle fractures. Data were analysed in our Computer Department. The sample comprised 348 patients with 444 fractures, and a male:female ratio of 2:1. Traffic accidents were the most common cause: 103 (41%) of the unilateral and 54 (56%) of the bilateral fractures, followed by alleged assault and falls. Low fractures were the most common -n = 314 of 444 (71%). The causes that involved considerable force (traffic accidents and falls) resulted in more dislocations of the mandibular head, more bilateral fractures, a tendency to fractures higher on the condyle and significantly more intracapsular fractures. Absence of molar occlusion also gave more high and fewer low fractures, but played no part in dislocation of the mandibular head from the glenoid fossa. PMID- 11010767 TI - Fractures of the mandibular condyle. Part 2: results of treatment of 348 patients. AB - This study was designed to record the results of conservative treatment of condylar fractures and to find out if there were any variables that were predictive of complications. Data were analysed in our computer department. During the period 1984-1996, all patients who presented with a fracture of the mandibular condyle and who attended for control examination one year after treatment were recorded at the end of treatment and one year later. The ability to open the mouth, deviation and occlusion were recorded. After one year 45 of the 348 patients (13%) had minor physical complaints such as reduced ability to open the mouth, deviation, or dysfunction. Ten of them (3%) had pain in the joint or muscles or both. Eight patients (2%) had malocclusion, which in seven could be related to dislocation of the condylar head out of the fossa. Five of the eight patients had had bilateral fractures. We conclude that conservative treatment of condylar fractures is non-traumatic, safe, and reliable and in only a few cases may cause disturbances of function and malocclusion. The risk associated with the latter is greatest with bilateral fractures and dislocation of the condylar head from the fossa. PMID- 11010768 TI - An investigation into the relationship between mandibular third molars and angle fractures in Nigerians. AB - We examined the relationship between the presence or absence of mandibular third molars and angle fractures in Nigerians between January 1976 and July 1997. Information was obtained from patients' case records and radiographs.A total of 490 patients were seen during the study period, 408 of whom (83%) had their third molars present, and 76 of whom had angle fractures. Road traffic accidents (n =304, 62%) were the main cause of the fractures. Of these, 44 (14%) had angle fractures compared with 14/75 (19%) patients in whom the mechanism of injury was a fight or assault.Sixty-five of the 408 patients whose third molars were present (16%) had angle fractures, as did 11/82 (13%) who did not have third molars (p =0.57). Of the 77 patients whose lower third molars were not erupted, 24 (31%) had angle fractures compared with 52/331 (16%) in whom the lower third molars were erupted (p =0.002). These findings suggest that angle fractures are uncommon in this population because of their particular mechanism of injury. In addition, the presence of a lower third molar does not necessarily predispose to fractures of the angle of the mandible. However, angle fractures are more likely to occur in people with unerupted lower third molars than in those in whom they have erupted. PMID- 11010769 TI - Ultrasound and fine needle aspiration cytology in the staging of neck lymph nodes in oral squamous cell carcinoma. AB - We prospectively studied 49 patients with oral squamous cell carcinoma (SCC) who had their cervical regional lymph nodes examined by ultrasound (US), with or without fine needle aspiration cytology (FNAC), for the detection of metastatic spread. They were screened over a two-year period (1993-1995) by computed tomography (CT) and magnetic resonance imaging (MRI) or both to measure the primary tumour. US, with or without FNAC, was used to stage the neck. All tumours were biopsied preoperatively and the diagnosis of SCC conformed by histological examination. Management of the neck was based on US staging. Thirty-five patients had neck dissections and 14 were managed conservatively. Mean follow-up was 30 months (range 24-48). All patients were alive at one year, but one died at 14 months from recurrence of disease in the neck. There were 3 false negatives (6%). We find that US, with or without FNAC, is an accurate (86%), sensitive (92%) and specific (83%) technique for the preoperative assessment of lymph node metastases in patients with SCC. PMID- 11010770 TI - Quantitative volume replacement in the correction of post-traumatic enophthalmos. AB - We describe a technique to correct enophthalmos that involves measurement of the volume of the injured orbit followed by an accurate replacement of this volume with an autogenous bone graft. Twelve consecutive patients were treated with this technique and all but one showed an improvement in enophthalmos. Four of the seven with diplopia improved after the operation. PMID- 11010771 TI - The bicoronal flap (craniofacial access): an audit of morbidity and a proposed surgical modification in male pattern baldness. AB - Maxillofacial surgeons have used the bicoronal flap for nearly three decades to gain access to the craniofacial skeleton. A retrospective analysis of 68 bicoronal flaps done over a five-year period showed that our incidence of permanent morbidity was low. Although 24 patients (35%) experienced some form of sensory abnormality immediately after the operation, this persisted for longer than two years in only one. Complete motor recovery occurred by one year in all 15 patients (22%) who developed postoperative frontalis weakness. Three patients developed male pattern baldness postoperatively, which resulted in exposure of the scar and poor cosmesis. This prompted a cadaveric study in which we assessed the feasibility of modifying the position of the standard bicoronal incision in people who are prone to hair loss. The pivotal point of the bicoronal flap was found to lie at its most inferior aspect. By extending the incision into the skin crease in front of the lobe of the ear it was possible to adjust the anteroposterior position of the bicoronal incision without limiting access to the facial skeleton. We therefore advocate occipitally placed incisions with preauricular extensions in patients who are prone to male pattern baldness. PMID- 11010772 TI - Incidence of human papillomavirus 16 and 18 infection and p53 mutation in patients with oral squamous cell carcinoma in Japan. AB - The purpose of this study was to investigate the prevalence of human papillomaviruses (HPVs) 16 and 18 infection, and p53 mutation in oral squamous cell carcinomas (SCCs) in Japanese patients. Our results showed a higher incidence of HPV16 and 18 infections than previous studies because we combined the findings of a consensus polymerase chain reaction (PCR), restriction fragment length polymorphism by using the restriction enzyme digestion of the PCR products and Southern blot hybridization. Each HPV16 and 18 E6/E7 DNA was detected in 9 (20%) and 25 (54%) of 46 samples. The p53 mutation in the exons from 5 to 8 were detected in 20 out of 46 samples (43%) by a PCR-single strand conformation polymorphism analysis. There was a significant relationship between HPV16 and the p53 mutation (P =0.02) suggesting that HPV16 infection has a mutagenic effect in oral SCC. However, neither HPV infection nor p53 mutation influenced survival. PMID- 11010773 TI - Grey-scale sonography in assessment of cervical lymphadenopathy: review of sonographic appearances and features that may help a beginner. AB - Ultrasound examinations of the neck in 218 patients with confirmed cervical lymphadenopathy were reviewed. Lymph nodes were assessed for their size, shape, internal architecture, echogenicity, nodal border, posterior enhancement, and ancillary features (adjacent soft tissues oedema, and matting). The hilus is a linear, echogenic, non-shadowing structure containing nodal vessels, and is continuous with fat around the node. Coagulation necrosis is an ill-defined, rounded, non-shadowing echogenic area within a node. It is less echogenic than the hilus and is not continuous with the fat around the node. Calcification is a highly echogenic focus within the node, which may be dense or punctate echogenic foci. It is not continuous with the fat around the node. Dense intranodal calcification usually produces shadowing. However, fine punctate calcification may not have posterior shadowing though, if the transducer frequency is increased, it may show thin lines. Cystic necrosis is focal, often ill-defined echolucent area within the node. Echogenicity of lymph nodes is usually compared with the adjacent muscles, and is classified as hypoechogenicity, isoechogenicity, and hyperechogenicity. The nodal border is assessed for its sharpness. Posterior enhancement is when the structures posterior to the node look more echogenic than neighbouring areas. Oedema of soft tissues is an ill defined, hypoechoic area around the node with loss of adjacent fascial planes. Nodes are considered matted when they are clumped or adherent to each other with no normal intervening soft tissue between them. Ultrasound features that help only in identifying abnormal nodes include size, shape, echogenic hilus, hypoechogenicity or isoechogenicity, echogeneity, coagulation necrosis, and a sharp nodal border. Ultrasound features that help to identify abnormal nodes as well as giving clues to the primary lesion include hyperechogenicity, intranodal calcification, intranodal cystic necrosis, ragged nodal border, posterior enhancement, adjacent soft tissue oedema, and matting. PMID- 11010774 TI - Tumour volume estimated by computed tomography as a predictive factor in carcinoma of the tongue. AB - This retrospective study evaluated tumour volume, estimated by computed tomography (CT), as a predictive factor in carcinoma of the tongue. Tumour volume was measured from pretreatment CT scans of 20 consecutive patients, followed up for at least 3 years, and this measurement was compared with tumour volume estimated from pathological specimens. T-stage and CT-derived tumour volume were compared with the clinical and pathological status of the nodes, and with the outcome of treatment. The measurement of tumour volume derived from CT correlated well with measurements derived from pathological examination, and tumour volume also predicted overall treatment failure. The disease-specific survival rate was 100% for patients with low-volume tumours (<13 cc) compared with 79% for those with stage T1 and T2 tumours.CT is a reliable way of measuring the volume of tumours in carcinoma of the tongue, and tumour volume is useful adjunct to the clinical tumour-node-metastases staging system. PMID- 11010776 TI - Role of surgical reduction of condylar fractures in the management of panfacial fractures. AB - We present nine panfacial fractures that were treated during a two-year period in which the first step in treatment was reduction of condylar fractures. There was no evidence of dental or skeletal alterations and measurement of the mandibular ramus and radiographic examination show that posterior facial height as well as projection and width of the inferior lower third of the face, was restored. The correct timing of surgical intervention and the use of rigid fixation allows the restoration of the morphological and functional nature of the face after panfacial fractures. PMID- 11010775 TI - Botulinum toxin: new treatment for temporomandibular disorders. AB - BACKGROUND: Temporomandibular disorders (TMDs) affect the face and jaws, and cause chronic pain and dysfunction in many people. As in other conditions involving the musculoskeletal system, controlling the myogenous component is an integral part of treatment. In this study, we evaluated subjective and objective responses to treatment with botulinum toxin A (BTX-A) in a group of 46 patients with TMDs. METHODS: 46 subjects with TMD were enrolled in this uncontrolled study and treated with BTX-A 150U. Both masseter muscles were injected with 50 U each and both temporalis muscles with 25 U each under electromyographic guidance. Subjects were assessed at two-week intervals for eight weeks. Outcome measures included subjective assessment of pain by visual analogue scale (VAS), measurement of mean maximum voluntary contraction (MVC), interincisal oral opening, tenderness to palpation, and a functional index based on multiple VAS. Medians of the data were taken for each outcome measure at each time point and subjected to Duncan's multiple range test. RESULTS: There were significant (P<0.05) differences in all median outcome measures between the pre-treatment assessment and the four follow-up assessments except for MVC. Although MVC was significantly reduced midway through the study, it had returned to pretreatment values by the final two assessments. All other outcome measures remained significantly different from the pretreatment findings. Paired correlation of variables including age, sex, diagnosis, depression index, and time of onset showed no significant differences. CONCLUSIONS: BTX-A injections produced significant improvements in pain, function, mouth opening, and tenderness to palpation. MVC initially diminished then returned to the initial values. Although the study was uncontrolled, the results strongly suggest that BTX-A reduces severity of symptoms and improves functional abilities for patients with TMD and that these extend beyond its muscle-relaxing effects. PMID- 11010777 TI - Staged operations for basal cell carcinoma of the face. AB - 211 patients (88 female, 123 male, mean age 63 years) presented with 279 basal cell carcinomas (191 primary tumours, 88 recurrent tumours) in the head and neck area that were excised. The resected margins were examined microscopically and the defect repaired at a second operation. The excision margin was 1-2 mm. The recurrence rate was 3%, which is lower than is achieved by other methods, and unaffected structures are optimally conserved. PMID- 11010778 TI - Assessing health outcomes after extraction of third molars: the postoperative symptom severity (PoSSe) scale. AB - A postoperative symptom severity (PoSSe) scale was devised from questions commonly used in the clinical assessment of patients who have had third molars extracted, and divided into subscales corresponding to seven main adverse effects that had been identified in a previous study. Ninety-seven patients completed the 15-item questionnaire at one week, and 71 patients at four weeks, after extraction of impacted third molars. After statistical testing, four items were discarded and the final PoSSe scale achieved a high level of internal reliability. Patients' PoSSe scores were highly correlated with scales that measured six distinct health dimensions. The level of social functioning, together with the number of impacted teeth and number of painkillers taken, explained 62% of the variation in PoSSe scores between patients one week postoperatively. The PoSSe was more responsive than the Short Form 36-item Health Survey. In conclusion, we found the PoSSe Scale to be a reliable, valid and responsive measure of the severity of symptoms after extraction of third molars, and of the impact of those symptoms on patients' perceived health. PMID- 11010779 TI - Secondary alveolar bone grafting in cleft lip and palate patients. AB - The case records of 72 patients in Northern Ireland who had secondary bone grafting of 94 alveolar clefts between 1987 and 1997 were examined. Details were recorded of age, sex, laterality of cleft, preoperative orthodontic treatment, whether grafting was combined with soft tissue revision, method of harvesting the graft, complications, eruption of canines, and the success of grafting as established by bone levels around the erupted canine. Bone levels were assessed on a four-point scale. Of 64 canines fully erupted in 94 clefts, 91% achieved bone levels of Type I or II and only two grafts failed. The outcome was more favourable when the canine was unerupted preoperatively. PMID- 11010780 TI - Impact of dedicated trauma lists on a maxillofacial surgical service. AB - In 1995, the maxillofacial surgical inpatient services in East Lancashire were centralized in Blackburn Royal Infirmary, and twice-weekly operating lists dedicated to maxillofacial trauma were established. We examined the non-elective workload for three-month periods before and after trauma lists became available to find out the proportion of non-elective operations done out of normal working hours. Although there was an increase in the total number of non-elective patients after centralization, day-time trauma lists allowed a reduction in the proportion of operations performed out of normal working hours. Fewer trauma cases were added to elective lists. Trauma lists allow the unit to comply with the recommendations of both NCEPOD and the Calman report, in that they maximize training opportunities for all staff and facilitate both audit and research. PMID- 11010781 TI - Isolated orbital blowout fractures: survey and review. AB - The management of orbital blowout fractures is controversial. The continuing debate includes the use of antibiotics and steroids, imaging, the surgical approach, and the choice of implant material for repair of the bony defect.A cross-sectional study was undertaken to assess current practice in treating orbital blowout fractures in the UK, in the form of a tick-box questionnaire. The questionnaire contained 9 closed and 2 open questions, and was forwarded to 256 practising fellows of the British Association of Oral and Maxillofacial Surgery. The response rate to the questionnaire was 73% (187/256). There was no consensus about the use of prophylactic antibiotics. However, 91% prescribed post-operative antibiotics and over half the respondents prescribed steroids. The most common imaging techniques used were computed tomography (CT, 88%) and plain radiograph (83%), 60% routinely sought an ophthalmic opinion and 65% assessed visual acuity. The most common surgical approaches were the subciliary (41%) and the infraorbital (37%), over half the respondents preferred to operate 6-10 days after the injury, and silicone elastomer was the preferred implant material of 66%. PMID- 11010782 TI - Irrigation of the Hall's air drill using a Foley catheter. PMID- 11010783 TI - Endogenous fatty acid synthesis in squamous cell carcinomas of the oral cavity. AB - We measured the synthesis of endogenous fatty acids in oral squamous cell carcinoma, its adjacent tissue, and normal oral tissues that were collected fresh from surgical specimens. The tissues were minced in Roswell Park Memorial Institute Medium 1640 (RPMI 1640) and incubated in [1(2)-(14)C] acetic acid, sodium salt ((14)CH(3)(14)CO(2)Na). Total lipids were then extracted and(14)C was measured by scintillation counter. We found that incorporation of(14)C in the total lipids of squamous cell carcinoma was significantly higher than in the other tissues. PMID- 11010784 TI - Analysis of tracheostomy-associated morbidity after operations for head and neck cancer. AB - Tracheostomy is a safe and effective way of securing the airway in patients who have radical resections for head and neck cancer. We audited the morbidity after 265 tracheostomies to identify the risks in relation to head and neck surgery, and to propose recommendations to improve care.Twenty-one tracheostomy-related complications were encountered in 256 patients (8%). Most complications occurred during the early postoperative period (72%). There were no tracheostomy-related deaths.Tracheostomies were retained for a median of 10 days (range 1-160). Delayed extubation was associated with extent of resection [P = 0.006], site of tumour (floor of mouth and anterior two thirds of tongue [P = 0.02]), and age (<61 years [P = 0.02]). Patients who were given preoperative radiotherapy were significantly more likely to develop a tracheostomy-related complication (P = 0.03). Patients with a tracheostomy complication were more likely to have other serious complications (P = 0.05) and in these patients there was a risk of delayed extubation (P = 0.06). We conclude that elective tracheostomy is essentially event-free, and most complications occur in the ward. PMID- 11010786 TI - Oral management of patients following oncology treatment: literature review. AB - Many tumours of the head and neck are treated by clinical oncologists using radiotherapy. This treatment modality is particularly effective in destroying rapidly dividing cells, hence its value in cancer treatment. The tissues of the oral mucosa, the salivary glands and blood vessels can be damaged as the result of radiotherapy treatment. As a result, patients experience unwanted oral effects that have both short and long-term implications. The outlook following treatment for head and neck cancer continues to improve and, as people keep their teeth into later life, dentists will increasingly be expected to address the oral problems that patients experience after radiotherapy. The aim of this article is to review the current literature concerning the implications and management of these oral implications of cancer treatment. PMID- 11010785 TI - Swallowing rehabilitation after oro-pharyngeal resection for squamous cell carcinoma. AB - The resection of even relatively small tumours that involve the base of tongue can lead to problems with swallowing. Free tissue transfer has improved the functional results and reduced the complications of head and neck surgery. The outcome after ablative operations depends on the site and extent of the resection, and in cases where the tongue base is involved, swallowing can be severely impaired. Aspiration in a patient with swallowing difficulties is of particular concern, however, there is a range of techniques that can be introduced during videofluoroscopy to reduce or eliminate aspiration. It is during the rehabilitation phase that procedures and techniques to improve swallowing and prevent aspiration should be incorporated. The efficacy of adaptive manoeuvres needs further evaluation, and in this study we aim to assess the effectiveness of compensatory procedures and therapy techniques (chin tuck and supraglottic swallow) in eliminating aspiration. We report on a study of the swallow function of 13 patients following surgical resection of the oropharynx including the base of tongue. Postoperative assessment by videofluoroscopy was carried out at 2 weeks, 1 month, 3 months, and 6 months according to a standard protocol. Subjects were analysed in two groups depending on the degree of resection of the tongue base resection (less than 1/4,1/4 or more).Patients' swallowing disorders were related to the extent of the resection and the consistency of the bolus. Those with involvement of a quarter of the tongue base or more generally had greater impairment, and radiotherapy tended to exacerbate these problems. Compensatory procedures and therapy techniques were effective in 50% of patients who aspirated, and tended to be more effective between the one month and 6 month follow-up in patients with smaller resections. PMID- 11010787 TI - Assessment of zygomatico-orbital complex fractures using ultrasonography. AB - Twenty-two patients were referred to the maxillofacial surgical unit for assessment and management of suspected fractures of the zygomatico-orbital complex. In each case, both routine plain radiographic and ultrasound examinations were made. The aim of the study was to investigate the sensitivity and reliability of ultrasound to detect such fractures. Imaging with ultrasound was carried out at five areas: the infraorbital margin; lateral wall of the maxillary sinus; zygomatic arch; frontozygomatic process; and orbital floor. Both radiographic and ultrasound findings were correlated with the findings at operation. An overall agreement of 85% between radiographs and ultrasound scans was found. Ultrasound imaging was most reliable at the lateral wall of the maxillary sinus, where the sensitivity was 94% and specificity 100%. The positive predictive value at this area was 100% compared with radiographic findings. We conclude that ultrasound is a useful tool in imaging facial trauma as an initial investigation, and can help to reduce the total number of radiographs required for the diagnosis of fractures of the zygomatico-orbital complex. PMID- 11010788 TI - Ultracision: the harmonic scalpel and its possible uses in maxillofacial surgery. AB - We report on the potential uses and benefits of a new development in incisive technology. The harmonic scalpel uses high-frequency mechanical energy to offer the surgeon controlled and precise incision and haemostasis. Vessels up to 2 mm in diameter may be sealed by coaptation with the blade before division. No special training or precautions are required before using this self-cleaning device. It produces considerably less smoke or smell than either diathermy or laser, which reduces the need for instrument exchanges and smoke evacuation. We think that it will be useful for tongue resections and possibly for neck dissections. PMID- 11010789 TI - Hemifacial atrophy with bilateral short roots. AB - We present a case in which the patient had both bilateral shortening of the roots, and hemifacial atrophy. As far as we know, this combination has not been described before. PMID- 11010790 TI - Amputation neuroma of the great auricular nerve after operations on the parotid gland. AB - We found an incidence of 6% (5/81) of traumatic neuroma after section of the great auricular nerve during operations on the parotid gland. Excision permits definitive diagnosis, the stump being allowed to retract beneath the belly of the sternomastoid muscle. However, excision is not always indicated, and the diagnosis can be made clinically allowing for a more conservative treatment policy. PMID- 11010791 TI - Fine needle aspiration biopsy (FNAB) for lesions of the salivary glands. AB - We examined 112 fine needle aspiration biopsy (FNAB) specimens of salivary glands (80 parotid and 32 submaxillary) taken between January 1989 and December 1995. Cytologic diagnoses were compared with the final histological diagnoses of the surgical specimens. The sensitivity and specificity were 84.8% and 93.7% respectively, and the accuracy was 91.1%. We conclude that FNAB by itself does not provide total security because of the high percentage of false-negatives. It is nevertheless useful when combined with an adequate clinical history, examination and radiological tests. PMID- 11010792 TI - Provision and quality assurance for head and neck cancer care. AB - In 1998, the British Association of Head and Neck Oncologists (BAHNO) published a report entitled 'Provision and quality assurance for head and neck cancer care in the United Kingdom. A nationally co-ordinated multidisciplinary approach'. This document considered, amongst other issues, operational policies for patient care. We audited three aspects of the care of 73 patients with head and neck cancer from initial referral to definitive treatment against standards set out in this report.Fifty-nine patients (81%) were seen within two weeks of initial referral, the maximum interval proposed by the BAHNO report. In 41 patients (56%), the diagnosis was confirmed and their treatment plan determined within the further two-week period suggested in the report. Of the 25 patients who required adjuvant radiotherapy, only 11 (44%) began treatment within six weeks of their surgical phase; this represented the greatest variance from the BAHNO recommendations. These findings highlight the need for units active in the treatment of specific diseases to audit their activity against available minimum standards in an attempt to improve the quality of patient care. PMID- 11010793 TI - Indication for epidural morphine for the relief of intractable pain in advanced oral cancer: report of four cases. AB - It can be difficult to manage the pain of advanced oral cancer. We present four patients in whom epidural morphine was used for intractable pain at primary or metastatic sites. For pain supplied by the trigeminal or cervical nerve a small dose of morphine was given through an epidural catheter inserted into the epidural space through C7-Th1. A favourable clinical response was achieved in three. In particular, in one patient who was given continuous morphine using a computerized ambulatory drug delivery system, we achieved excellent efficacy and stable control of pain. We think that the effect of the epidural morphine was decreased in the patient who did not respond because he had previously been treated with high oral doses. The present study confirmed that morphine given epidurally in small doses has a strong and prolonged analgesic action with less toxicity than when given orally. PMID- 11010794 TI - Differential diagnosis of calcium pyrophosphate dihydrate deposition of the temporomandibular joint. AB - Calcium pyrophosphate dihydrate (CPPD) deposition disease (pseudogout) of the temporomandibular joint (TMJ) is rare. It is characterized by the presence of crystal deposits that are birefringent under polarized light. Although these crystals are characteristically weakly birefringent, some other crystals such as those of calcium oxalate, synthetic steroids, and ethylenediaminetetraacetic acid are also birefringent. The differential diagnosis should therefore be based on a quantitative analysis of crystals or observation of the crystal structure in calcified sections. We present a case of CPPD deposition disease of the TMJ and report on the value of such an analysis to substantiate the diagnosis. PMID- 11010795 TI - True eminectomy for internal derangement of the temporomandibular joint. AB - We carried out a prospective study to assess the short- and long-term outcomes of true eminectomy in 20 consecutive patients with type III or IV anteriorly displaced discs in whom conventional conservative treatment had failed to resolve the symptoms. Maximum opening was measured preoperatively, 3-6 months postoperatively and at one year postoperatively. Each patient filled out a Mandibular Functional Impairment Questionnaire [MFIQ] and Clinical Dysfunctional Index both preoperatively and one year postoperatively. Imaging records included preoperative and one year postoperative orthopantomograph (OPG) and magnetic resonance imaging (MRI). Maximum mouth opening one year postoperatively had increased in 95% of patients (range 5-20 mm, mean 12 mm). There was an improvement in symptoms, using the MFIQ, in 85% of patients. The Clinical Dysfunctional Index was reduced to Type I in most patients. MRI showed increase in rotation and translatory movement of the condylar head. We conclude that true eminectomy may be used to successfully treat type III and type IV anteriorly displaced discs in patients who have failed to respond to conventional conservative treatment. PMID- 11010796 TI - Vascular osteomuscular autograft prefabrication using coral, type I collagen and recombinant human bone morphogenetic protein-2. AB - In 20 male Sprague-Dawley rats, 10 pieces of L-shaped coral combined with type I collagen and recombinant human bone morphogenetic protein-2 (rhBMP-2) and 10 discs (diameter 5 mm) were wrapped in the gracilis muscle, and left pedicled on the femoral vessels. Untreated coral was buried in muscle at a distant control site in 4 animals. After 3 weeks, autografts were examined for the shape of new bone, vascular patency, and induction of bone. In all grafts in viable tissue, heterotopic bone was formed. The shape of the new bone was the same as that of the coral, and there was no significant inflammatory reaction. Part of the coral in the composite was absorbed. Bone was not formed in any of the control sites. Coral and type 1 collagen are effective as a carrier for BMP to prefabricate vascular osteomuscular autografts with designed shape. There is a potential clinical application for BMP to bioengineer microvascular free flaps with intrinsic skeletal muscle for maxillofacial reconstruction. PMID- 11010797 TI - Composite autogenous bone and demineralized bone matrices used to repair defects in the parietal bone of rabbits. AB - We compared the amount of new bone produced by endochondral and intramembranous autogenous bone grafts in the presence of demineralized bone matrices (DBMs) prepared from intramembranous bone (DBM(IM)) or endochondral bone (DBM(EC)). Thirty-five bone defects were created in the parietal bone of 20 New Zealand White rabbits. In the experimental groups, 5 defects were grafted with endochondral bone, 5 with endochondral bone mixed with DBM(IM)) (EC-DBM(IM)), 5 with intramembranous bone mixed with DBM(IM)(IM-DBM(IM)) and 6 with endochondral bone mixed with DBM(EC)(EC-DBM(EC)). In the control groups, 10 defects were left alone (passive control) and 4 were grafted with rabbit skin collagen (active control). They were all killed on day 14 and the defects were prepared for histological study. Serial sections were cut across the whole defect. Quantitative analyses were made on 202 sections of the experimental groups by image analysis. A total of 414%, 708%, and 85% more new bone was formed in defects grafted with composite EC-DBM(IM), IM-DBM(IM)and EC-DBM(EC), respectively, than those grafted with endochondral bone alone (P<0.001). No bone was formed in either passive or active controls. In conclusion, demineralized bone matrices, particularly those derived from intramembranous bone, have extremely high osteoinductive properties and greatly improve the integration of autogenous bone grafts in the skull. PMID- 11010798 TI - Unilateral temporalis muscle hypertrophy managed with botulinum toxin type A. PMID- 11010799 TI - First symptom of multifocal skeletal tuberculosis in children. PMID- 11010800 TI - Re: Kerawala C.J. Oral cancer, smoking and alcohol: the patients' perspective. Br J Oral Maxillofac Surg 1999; 37: 374-376. PMID- 11010801 TI - Re: Newton J.P. et al. Masseteric hypertrophy?: preliminary report. Br J Oral Maxillofac Surg 1999; 37: 405-408. PMID- 11010802 TI - Third molar surgery. PMID- 11010803 TI - Caution in use of bicortical intermaxillary fixation screws. PMID- 11010805 TI - Notch signaling: a dance of proteins changing partners. PMID- 11010806 TI - Fas-mediated apoptosis of proliferating, transiently growth-arrested, and senescent normal human fibroblasts. AB - Previous studies suggest that apoptotic signaling may require proteins that are critical to cellular proliferation and cell cycle regulation. To further examine this question, proliferating, transiently growth-arrested, and senescent normal human fibroblasts were induced to undergo apoptosis in response to two distinct mediators of apoptosis-Fas (APO-1/CD95) death receptor and staurosporine. Ligation of the Fas receptor in the presence of cycloheximide or actinomycin D resulted in apoptosis of proliferating cells, cells transiently growth arrested by gamma-irradiation or serum starvation (i.e., G(0) arrest), and permanently growth-arrested senescent fibroblasts. Proliferating and G(0)-arrested cells were also susceptible to staurosporine-mediated apoptosis. Surprisingly, gamma irradiated cells did not undergo staurosporine-mediated apoptosis, and remained viable for a prolonged time. Fas-mediated apoptosis of senescent fibroblasts was evidenced by chromosome condensation and by activation of caspase-8 and -3, proteases crucial for the execution of the Fas apoptosis pathway. In addition, ligation of the Fas receptor in G(0)-arrested cells did not result in the activation of p34(cdc2) kinase, arguing that activation of this kinase is not essential in this apoptotic process. From these studies we conclude that proliferating, transiently growth-arrested, and senescent normal human fibroblasts are susceptible to apoptotic signals and that apoptosis is not necessarily dependent upon cell cycle or proliferative state of the cell. PMID- 11010807 TI - Thrombin inhibits apoptosis of monocytes and plasminogen activator inhibitor 2 (PAI-2) is not responsible for this inhibition. AB - Plasminogen activator inhibitor 2 (PAI-2) has been shown to inhibit apoptosis in transfected cells. We have investigated this phenomenon in activated human monocytes, which are a physiological source of intracellular PAI-2. Apoptosis of monocytes was rapidly induced by removal of serum, addition of hydrogen peroxide, or binding of a monoclonal antibody to Fas. Treatment of monocytes with thrombin or lipopolysaccharide (LPS) inhibited apoptosis of monocytes and also up regulated intracellular PAI-2. Increased apoptosis was accompanied with increased activity of caspases 3 and 8. Thrombin or LPS treatment of monocytes decreased the activity of both caspases, which correlated with protection from apoptosis. The role for PAI-2 in protection of monocytes from apoptosis was studied. Monocytes were transfected with antisense oligonucleotides that blocked PAI-2 antigen, and antisense for PAI-2 had no effect on apoptosis of monocytes. No interaction was evident between PAI-2 and recombinant caspases 3 and 8 in vitro. PAI-2 was not a substrate for caspases during apoptosis of monocytes, although some cleavage of recombinant PAI-2 by caspase 3 was evident in vitro. This study shows that thrombin or LPS protected monocytes from apoptosis and that PAI-2 did not mediate this inhibitory effect. PMID- 11010808 TI - Immunoelectron microscopy provides evidence that tumor necrosis factor receptor associated protein 1 (TRAP-1) is a mitochondrial protein which also localizes at specific extramitochondrial sites. AB - The tumor necrosis factor receptor-associated protein 1 (TRAP-1) interacts with a variety of proteins involved in diverse functions. We have used quantitative immunogold electron microscopy and biochemical analysis to evaluate the subcellular distribution of TRAP-1 in rat tissues. Immunofluorescence employing a polyclonal antibody raised to human recombinant TRAP-1 reveals specific staining of mitochondria and nuclear region in mammalian cells. Western blot analysis of purified rat liver mitochondrial subfractions with the TRAP-1 antibody reveals that the cross-reactive protein (M(r) approximately 80 kDa) is mainly present in the matrix compartment. Immunogold labeling of rat tissue sections embedded in LR Gold resin shows strong labeling of mitochondria in all the tissues examined (viz., liver, heart, pancreas, kidney, spleen, anterior pituitary gland). Additionally, specific and significant labeling with TRAP-1 antibody was also observed in certain tissues in a number of nonmitochondrial locations, including pancreatic zymogen granules, insulin secretory granules, cardiac sarcomeres, and nuclei of pancreatic and heart cells, and on the cell surface of blood vessel endothelial cells. Western blot analysis showed that a cross-reactive protein of similar molecular mass as TRAP-1 is present in purified pancreatic zymogen granules. Immunogold labeling was prevented in all tissues by preadsorption of the TRAP-1 antibody with the purified recombinant TRAP-1 protein. These observations and the fact that TRAP-1 is synthesized with a typical mitochondrial targeting presequence strongly indicate that TRAP-1 is primarily a mitochondrial matrix protein. The localization of this protein at specific extramitochondrial sites raises interesting and fundamental questions regarding the possible mechanisms by which these proteins are translocated to such sites. PMID- 11010809 TI - Site-directed mutagenesis and deletion of three phosphorylation sites of calsequestrin of skeletal muscle sarcoplasmic reticulum. Effects on intracellular targeting. AB - Calsequestrin (CS) is segregated to the junctional sarcoplasmic reticulum (jSR) of skeletal muscle fibers and is responsible for intraluminal Ca(2+) binding. A chimeric CS-HA1, obtained by adding the nine-amino-acid viral epitope hemagglutinin (HA1) to the carboxy-terminal of CS and shown to be correctly segregated to skeletal muscle jSR in vivo (A. Nori, K. A. Nadalini, A. Martini, R. Rizzuto, A. Villa, and P. Volpe, 1997, Am. J. Physiol. 272, C1420-C1428), is mutagenized in order to identify domains of CS involved in targeting. Since a putative targeting mechanism of CS implies phosphorylation-dependent steps in the endoplasmic reticulum (ER) and/or Golgi complex, five CS-HA1 mutants disrupting the three phosphorylation sites of CS (Thr(189), Thr(229), and Thr(353)) were engineered by either site-directed mutagenesis or deletion: CS-HA1DeltaP1 (Thr(189) --> Ile); CS-HA1DeltaP2 (Thr(229) --> Asn); CS-HA1DeltaP1,2; in which Thr(189) and Thr(229) were changed to Ile and Asn, respectively; and CS HA1Delta14(COOH) and CS-HA1Delta49 (COOH), in which 14 residues (Glu(354) Asp(367)) and 49 residues (Asp(319)-Asp(367)), respectively, were deleted at the carboxy-terminal. Mutant cDNAs were transiently transfected in either HeLa cells, cultured myoblasts of rat skeletal muscle, or regenerating soleus muscle fibers of adult rats. Each CS-HA1 mutant was identified by Western blot as a single polypeptide of the predicted molecular weight. The intracellular localization of CS-HA1 mutants was studied by immunofluorescence using specific antibodies against either CS or HA1. CS-HA1 mutants colocalized with ER markers, e.g., calreticulin, and partially overlapped with Golgi complex markers, e.g., alpha mannosidase II, in HeLa cells and myotubes. CS-HA1 mutants were expressed and retained in ER and ER/SR of HeLa cells and myotubes, respectively, and correctly segregated to jSR of regenerating soleus muscle fibers. Thus, the targeting mechanism of CS in vivo is not affected by phosphorylation(s); i.e., sorting and segregation of CS appear to be independent of posttranslational phosphorylation(s). PMID- 11010810 TI - Effector mechanisms of fenretinide-induced apoptosis in neuroblastoma. AB - Fenretinide is an effective inducer of apoptosis in many malignancies but its precise mechanism(s) of action in the induction of apoptosis in neuroblastoma is unclear. To characterize fenretinide-induced apoptosis, neuroblastoma cell lines were treated with fenretinide and flow cytometry was used to measure apoptosis, free radical generation, and mitochondrial permeability changes. Fenretinide induced high levels of caspase-dependent apoptosis accompanied by an increase in free radicals and the release of cytochrome c in the absence of mitochondrial permeability transition. Apoptosis was blocked by two retinoic acid receptor (RAR)-beta/gamma-specific antagonists, but not by an RARalpha-specific antagonist. Free radical induction in response to fenretinide was not blocked by the caspase inhibitor ZVAD or by RAR antagonists and was only marginally reduced in cells selected for resistance to fenretinide. Therefore, free radical generation may be only one of a number of intracellular mechanisms of apoptotic signaling in response to fenretinide. These results suggest that the effector pathway of fenretinide-induced apoptosis of neuroblastoma is caspase dependent, involving mitochondrial release of cytochrome c independently of permeability changes, and mediated by specific RARs. As the mechanism of action of fenretinide may be different from other retinoids, this compound may be a valuable adjunct to neuroblastoma therapy with retinoic acid and conventional chemotherapeutic drugs. PMID- 11010811 TI - Inhibition of cell growth and apoptosis by inducible expression of the transcriptional repressor Mad1. AB - Mad1 is a Myc antagonist that heterodimerizes with Max and functions as a transcriptional repressor. We have studied the effects of Mad1 on cell growth, cell cycle distribution, and apoptosis using Mad1-inducible cell lines. Expression of Mad1 inhibited cell proliferation, S-phase entry, and colony formation, changes that were accompanied by a reduction in CDK2 activity. The inhibition of Mad1 on cell proliferation was potentiated by serum starvation and was paralleled by accumulation of cells in the G0/G1 and the G2 phases of the cell cycle. Mad1 also reduced apoptosis induced by serum withdrawal and by the cytostatic drug cisplatinum. The effects on both cell growth and apoptosis were dependent on the mSin3 interaction domain of Mad1, which is necessary for recruitment of histone deacetylases and corepressors, suggesting that transcriptional repression is mediating these functions. Taken together with the expression pattern of Mad1, these results suggest that Mad1 plays an important role during initiation of differentiation by inhibiting cell proliferation and blocking apoptosis. PMID- 11010812 TI - Differential induction of gelatinase B (MMP-9) and gelatinase A (MMP-2) in T lymphocytes upon alpha(4)beta(1)-mediated adhesion to VCAM-1 and the CS-1 peptide of fibronectin. AB - Integrin alpha(4)beta(1) on the surface of T lymphocytes interacts with vascular cell adhesion molecule-1 (VCAM-1) and fibronectin during migration of lymphocytes from the blood to sites of inflammation. Migrating lymphocytes actively modify their environment through a number of mechanisms including proteolysis of the extracellular matrix by matrix metalloproteinases (MMP) synthesized by the cells. In this study, expression of MMP upon alpha(4)beta(1)-mediated adhesion of leukocytes to two major ligands, the IIICS-1 domain of fibronectin and VCAM-1, has been examined. Adhesion of T lymphoblastoid Jurkat cells to the CS-1 peptide induced expression of mRNA for two MMPs, gelatinase A (MMP-2) and gelatinase B (MMP-9). As evaluated by relative RT-PCR and Northern blot analyses, the level of mRNA was upregulated about 4- to 5-fold for both MMPs compared to control cells maintained in suspension. With time, both enzymes were detected in conditioned media and inside the cells, and their identities were verified by Western blotting and gelatin zymography. Adhesion of Jurkat cells to the second major alpha(4)beta(1) ligand, VCAM-1, upregulated mRNA for MMP-2 (3.5-fold) and failed to induce expression of mRNA for MMP-9. Accordingly, only MMP-2 protein was detected in conditioned media of cells adherent to VCAM-1. Occupancy of alpha(4)beta(1) on the surface of suspended cells with soluble CS-1 peptide or VCAM-1 did not upregulate synthesis and release of MMPs. A similar pattern of induction of MMPs after adhesion to CS-1 and VCAM-1 was observed in T lymphocytes isolated from human blood. These results demonstrate that adhesion of T lymphocytes through alpha(4)beta(1) to different ligands, which bind to similar or overlapping sites in the integrin, induces intracellular events leading to distinct patterns of MMPs biosynthesis. PMID- 11010813 TI - The whole meiotic process can occur in vitro in untransformed rat spermatogenic cells. AB - The aim of the present study was to assess whether the whole meiotic process of spermatogenic cells is able to take place in vitro. Fragments of seminiferous tubules from 20- to 22- or 28-day-old rats were seeded in medium containing 0.2% fetal calf serum in bicameral chambers and then cultured for 4 weeks in a chemically defined medium. The differentiation of meiotic germinal cells was followed by four criteria: (i) ultramicroscopic examination of the different types of germ cells present in the cell layer throughout the culture period; (ii) determination of the changes in DNA content per nucleus of the cell population seeded with time in culture; (iii) assessment of the ability of germinal cells to transcribe genes expressed after completion of meiosis; and (iv) monitoring the fate of BrdU-labeled leptotene spermatocytes. The ultrastructural study showed that the overall organization of the cells in the culture well recalls that of the seminiferous epithelium throughout the culture period. Moreover the identification of young round spermatids 21 days after seeding suggested that these spermatids had been formed very recently in culture. Determination of DNA content per nucleus showed that a 1C cell population could be observed after several days of cultures reaching 6 to 10% of total cells. An exponential-like increase in the amounts of the mRNAs encoding for TP1 or TP2 occurred from the time when 1C cells appeared in the culture until the end of the experiment. Finally, BrdU-labeled leptotene spermatocytes differentiated into pachytene spermatocytes and then into secondary spermatocytes, and BdrU-labeled round spermatids were observed from Day 21 of culture onward. Taken together these results indicate that the whole meiotic process from leptotene spermatocyte to round spermatid can indeed occur in vitro under the present culture conditions. PMID- 11010814 TI - 14-3-3 protein family members have a regulatory role in retinoic acid-mediated induction of cytokeratins in F9 cells. AB - We have found that the expression of five 14-3-3 protein isoforms is induced during the retinoic acid (RA)-mediated differentiation of mouse embryonal carcinoma F9 cells. The induced expression of the 14-3-3 proteins is presumed to have a role in enhancing the mitogen-activated protein kinase (MAPK) activity during RA-mediated F9 cell differentiation, because using genetically engineered budding yeast we showed that these isoforms enhanced the signaling in the MAPK cascade mainly through the interaction with Raf-1. Then we assessed the role of increased MAPK activity in F9 cell differentiation by interfering with signaling in the MAPK cascade in F9 cells. The exogenous expression of dominant-negative MEK1 efficiently abrogated RA-mediated induction of the cytokeratins EndoA and EndoC in the F9 cells. These results suggest that the 14-3-3 proteins play a role in the efficient induction of the cytokeratins during F9 cell differentiation through their signal enhancing activity in the MAPK cascade. PMID- 11010815 TI - A conformational change in PBX1A is necessary for its nuclear localization. AB - The fly homeodomain (HD) protein EXTRADENTICLE (EXD) is dependent on a second HD protein, HOMOTHORAX (HTH), for nuclear localization. We show here that in insect cells the mammalian homolog of EXD, PBX1A, shows a similar dependence on the HTH homologs MEIS1, 2, and 3 and the MEIS-like protein PREP1. Paradoxically, removal of residues N-terminal to the PBX1A HD abolishes interactions with MEIS/PREP but allows nuclear accumulation of PBX1A. We use deletion mapping and fusion to green fluorescent protein to map two cooperative nuclear localization signals (NLSs) in the PBX HD. The results of DNA-binding assays and pull-down experiments are consistent with a model whereby the PBX N-terminus binds to the HD and masks the two NLSs. In support of the model, a mutation in the PBX HD that disrupts contact with the N-terminus leads to constitutive nuclear localization. The HD mutation also increases sensitivity to protease digestion, consistent with a change in conformation. We propose that MEIS family proteins induce a conformational change in PBX that unmasks the NLS, leading to nuclear localization and increased DNA binding activity. Consistent with this, PBX1 is nuclear only where Meis1 is expressed in the mouse limb bud. PMID- 11010816 TI - Centrosome-specific perturbations during in vitro maturation of mouse oocytes exposed to cocaine. AB - Previous studies indicating that cocaine may perturb meiotic chromosome segregation in mammalian oocytes prompted an analysis of the effects of cocaine on mouse oocytes matured in vitro under defined exposure conditions. Cumulus enclosed mouse oocytes were matured in vitro in the continuous presence of cocaine and assessed for meiotic cell cycle progression and centrosome microtubule organization using a combination of cytogenetic and fluorescence microscopic techniques. Both of these approaches demonstrated that cocaine had little effect on meiotic cell cycle progression to metaphase of meiosis-2 except at the highest dose tested (1000 microg/ml) where progression from metaphase-1 to metaphase-2 was inhibited. Cytogenetic analyses further showed that bivalent segregation was moderately affected and the incidence of premature centromere separation was significantly decreased following cocaine treatment. Under conditions of cocaine exposure, striking changes in meiotic spindle structure and cytoplasmic centrosome organization were observed. A 36% reduction in spindle length was associated with a loss of nonacetylated microtubules and fragmentation of spindle pole centrosomes. Moreover, in oocytes exposed to cocaine during maturation, a doubling in cytoplasmic centrosome number was observed. These results are discussed with respect to the relative roles of chromosomes and centrosomes in establishing and maintaining functional microtubule organization during meiosis in oocytes. PMID- 11010817 TI - Growth state-dependent binding of USF-1 to a proximal promoter E box element in the rat plasminogen activator inhibitor type 1 gene. AB - Induced PAI-1 gene expression in renal epithelial (NRK-52E, clone EC-1) cells occurs as part of the immediate-early response to serum. PAI-1 transcripts are maximally expressed early in G(1) (within 4 h of serum addition to quiescent EC-1 cells) and then subsequently decline to basal levels prior to entry into DNA synthetic phase. Comparative analysis of PAI-1 mRNA abundance and de novo synthesized thiolated RNA in quiescent cells, as well as at 4 h (early G(1)) and 20 h (late G(2)) postserum addition, in conjunction with RNA decay measurements indicated that PAI-1 gene regulation upon growth activation was predominantly transcriptional. An E box motif (CACGTG), important in the induced expression of some growth state-dependent genes, mapped to nucleotides -160 to -165 upstream of the transcription start site in the PAI-1 proximal promoter. Mobility-shift assessments, using a 18-bp deoxyoligonucleotide construct containing the E box within the context of PAI-1-specific flanking sequences, confirmed binding of EC 1 nuclear protein(s) to this probe and, specifically, to the E box hexanucleotide site. The specificity of this protein-probe interaction was verified by competition analyses with double-stranded DNA constructs that included E box deoxyoligonucleotides with non-PAI-1 flanking bases, mutant E box sequences incapable of binding NRK nuclear proteins, and unrelated (i.e., AP-1) target motifs. Extract immunodepletion and supershift/complex-blocking experiments identified one PAI-1 E box-binding protein to be upstream stimulatory factor-1 (USF-1), a member of the HLH family of transcription factors. Mutation of the CACGTG site to TCCGTG in an 18-bp PAI-1 probe inhibited the formation of USF-1 containing complexes confirming that an intact E box motif at -160 to -165 bp in the PAI-1 promoter and, in particular, the CA residues at -165 and -164 are essential for USF-1 binding. Incorporation of this 2 bp change into a reporter construct containing 764 bp of the proximal PAI-1 "promoter" ligated to a CAT gene effectively reduced (by 74%) CAT activity in cycling cells. An intact E box motif at nucleotides -160 to -165 in the PAI-1 promoter, thus, is an important functional element in the regulation of PAI-1 transcriptional activity in renal cells. PMID- 11010818 TI - Epidermal growth factor receptor efficiently activates mitogen-activated protein kinase in HeLa cells and Hep2 cells conditionally defective in clathrin-dependent endocytosis. AB - Epidermal growth factor (EGF)-induced signaling was investigated in cells conditionally defective in clathrin-dependent endocytosis by overexpression of K44A dynamin in HeLa cells and potassium depletion in Hep2 cells. Overexpression of mutant dynamin disrupts high-affinity EGF-EGF receptor (EGFR) interaction (T. Ringerike, E. Stang, L. E. Johannessen, D. Sandnes, F. O. Levy, and I. H. Madshus, 1998, J. Biol. Chem. 273, 16639-16642). However, the EGFR substrates Shc and c-Cbl were as efficiently tyrosine phosphorylated in endocytosis-deficient HeLa cells exhibiting only low-affinity EGFRs as in HeLa cells with intact endocytosis and with both high- and low-affinity EGFRs. Both Raf and mitogen activated protein kinase (MAPK) were activated to the same extent and with the same kinetics. HeLa cells distributed equally in the cell cycle regardless of EGFR internalization. Upon potassium depletion of Hep2 cells, EGF-induced EGFR endocytosis was inhibited. However, the EGFR and MAPK were efficiently activated by EGF in both the absence and the presence of clathrin-dependent endocytosis. The EGFR was weakly tyrosine phosphorylated by potassium depletion even in the absence of EGF, and this activation resulted in detectable activation of MAPK. Our results demonstrate that internalization of EGFR by clathrin-dependent endocytosis is not required for activation of MAPK. PMID- 11010819 TI - Stem cell characteristics of transplanted rat mammary clonogens. AB - Rat mammary glands contain a subpopulation of clonogenic epithelial cells with large proliferation and differentiation potentials. When transplanted, the clonogens in monodispersed rat mammary epithelial cell suspensions give rise to either alveolar units (AUs) or ductal units (DUs) depending on the nature of the hormonal milieu in the graft recipient. Clonogens are also the primary cells of origin of mammary cancer following exposure to ionizing radiation or chemical carcinogens. Given the other stem cell characteristics of mammary clonogens, it would be expected that the primary AUs and DUs to which they give rise when grafted and hormonally stimulated (a) would be derived from the same clonogenic cell subpopulation, (b) would contain all of the functionally differentiated cell types of homologous parts of comparably stimulated mammary glands in situ, and (c) would also contain clonogen subpopulations capable when subtransplanted of giving rise to secondary AUs and DUs of similar cell composition. The current experiments were designed to test these expectations. The data are discussed in the context of results of previous studies with this and other experimental models. The results further support the conclusion that rat mammary clonogens are multipotent mammary stem cells. PMID- 11010820 TI - Effects of antiandrogens on chromatin remodeling and transcription of the integrated mouse mammary tumor virus promoter. AB - Inhibition of the ligand-activated androgen receptor (AR) by antiandrogens plays an important role in the treatment of various hyperandrogenic disorders including prostate cancer. However, the molecular mechanisms of antiandrogen activity in vivo remain unclear. In this study we analyzed the effects of cyproterone acetate (CPA), flutamide (F), and hydroxyflutamide (OHF) on transcriptional activation and chromatin remodeling of the genomically integrated mouse mammary tumor virus (MMTV) promoter. This promoter has provided an excellent model system to study the impact of steroid hormones on transcriptional activation in the context of a defined chromatin structure. The MMTV hormone response element is positioned on a phased nucleosome, which becomes remodeled in response to steroids. We utilized this model system in mouse L-cell fibroblasts that contain a stably integrated MMTV promoter. In these cells, dihydrotestosterone (DHT) induced a large increase of AR protein levels that correlated with transcriptional activation and chromatin remodeling of the MMTV promoter. Coadministration of DHT and CPA or DHT and OHF in these cells inhibited the increase of AR levels, which resulted in a strong blockage of transcriptional activation and chromatin remodeling of the MMTV promoter. In contrast, F had no significant influence on these activities. We conclude that a major portion of the antiandrogenic effects of CPA and OHF in vivo are mediated by the reduction of AR levels. PMID- 11010821 TI - Homocysteine inhibits retinoic acid synthesis: a mechanism for homocysteine induced congenital defects. AB - Hyperhomocysteinemia is frequently associated with congenital defects of the heart and neural tube and is a suspected pathogenic factor in atherosclerosis and neoplasia. Results in the present report show homocysteine treatment disrupts normal development of avian embryos; and this effect is prevented by retinoic acid. Based on this, we hypothesize that homocysteine may exert its teratogenic effects by disrupting retinoic acid signaling during development. A reporter cell line transfected with a retinoic acid response element (RARE) linked to a lacZ reporter gene was used to identify the site of retinoid inhibition. Using this reporter cell line, we show that homocysteine inhibits the oxidation of retinal to retinoic acid with concentrations of homocysteine that are in the pathophysiological range (.05 to 0.5 mM). In contrast, homocysteine concentrations as high as 5 mM are unable to inhibit the induction of lacZ by retinoic acid. We show that cellular uptake of homocysteine is sensitive to the specific L-system transport inhibitor, bicycloheptane, and bicycloheptane blocks the inhibition of retinoic acid synthesis by homocysteine, demonstrating that this inhibition occurs intracellularly. These results suggest that homocysteine induced congenital defects are due to the specific ability of homocysteine to inhibit conversion of retinal to retinoic acid. PMID- 11010822 TI - Inactivation of Ca(2+) action potential channels by the MEK inhibitor PD98059. AB - In a previous study involving the inhibition of mitogen-activated protein kinase (MAPK) activation during fertilization of the marine worm Urechis caupo, we found that PD98059, but not U0126, caused multiple sperm penetrations in oocytes (Gould and Stephano, 1999, Dev. Biol. 216, 348-358). Since these oocytes are protected against polyspermy by a positive shift in membrane potential at fertilization (Gould-Somero et al., 1979, J. Cell Biol. 82, 426-440), we investigated the effects of PD98059 on the electrical properties of the oocyte membrane. PD98059, but not U0126, selectively blocked the voltage-dependent Ca(2+) channels that participate in the electrical polyspermy block. We also noted previously that PD98059 had more serious effects than U0126 on chromosome behavior during meiosis. This, too, could be explained by the effect on Ca(2+) channels, since when U0126-treated eggs were fertilized in low Ca(2+) seawater to reduce Ca(2+) uptake, similar effects were produced. These results show that PD98059 has side effects unrelated to the inhibition of MAPK activation and underscores the need for caution in interpreting the results of experiments with this widely used MEK inhibitor. PMID- 11010823 TI - Immunology of bacille Calmette-Guerin and related topics. PMID- 11010824 TI - Preventing tuberculosis with bacillus Calmette-Guerin vaccine: a meta-analysis of the literature. AB - This article reviews a previously published meta-analysis of 1264 titles or abstracts and 70 selected studies for evaluation of the efficacy of bacillus Calmette-Guerin (BCG) vaccine in preventing tuberculosis. Following review, data from 26 studies were included in the analysis. These 26 studies reveal that vaccination with BCG significantly reduces the risk of tuberculosis by an average of 50%. This level of protection persists across a number of subgroups defined by age at vaccination and study design. Vaccination with BCG was significantly associated with a reduction in the incidence of pulmonary tuberculosis and extrapulmonary disease. In general, the results of this meta-analysis lend weight and confidence to arguments favoring the use of BCG vaccine. PMID- 11010825 TI - Animal models for experimental tuberculosis. AB - Animal models for tuberculosis vaccine assays have evolved through a series of sequential experiments aimed at optimizing the activity of a particular vaccine product. As a result, studies that use different animal models do not agree on the potency ranking of antituberculosis vaccines, and each major test-system variable contributes to the disagreement. Disagreements among laboratories about the efficacy of vaccines are in part due to differences in test systems. A survey of potency assays of tuberculosis vaccines suggests that, based on the choice of a specific combination of variables in the test model, an investigator can show that any given vaccine product has superior potency. In view of these problems with animal models for research on tuberculosis vaccines, we recommend that attention should be focused on those animal models that replicate the key aspects of the natural history of human tuberculosis. The development of vaccines by means of new technologies requires animal models to assay the protective potency of vaccines that, ideally, inhibit tubercle bacilli at the point of infection. The development of new tuberculosis vaccines may be handled most efficiently in a joint venture that includes both laboratories involved in vaccine production and laboratories concerned with animal models. PMID- 11010826 TI - What does tuberculin reactivity after bacille Calmette-Guerin vaccination tell us? AB - The effect of bacille Calmette-Guerin (BCG) vaccination on tuberculin reactivity is briefly reviewed. BCG vaccination will almost invariably result in tuberculin conversion with a positive tuberculin skin test developing 4-8 weeks after vaccination. However, these tuberculin reactions will wane-rapidly in all individuals who receive the vaccine in the neonatal period and more slowly in those who are vaccinated at an older age such as during the primary-school years. Of BCG vaccine recipients whose initial tuberculin skin test is negative, 10%-25% will have a positive tuberculin skin test if they are retested within 1-4 weeks the so-called "booster phenomenon. " There is no relationship between tuberculin reactivity after BCG vaccination and the protective efficacy of the vaccine against development of active tuberculosis. Therefore, the ideal BCG vaccine would produce a scar at the site of injection to identify individuals who have been vaccinated but would have no effect on tuberculin reactivity. PMID- 11010827 TI - Management of adverse reactions to bacille Calmette-Guerin vaccine. AB - Published reports appear to underestimate the true rate of adverse reactions to bacille Calmette-Guerin (BCG) vaccine. At a recent national conference on tuberculosis control among aboriginal populations, lack of awareness of what constitutes an adverse reaction was considered a possible contributing factor to underreporting. The following review defines a normal BCG response and discusses the management of complications when they occur. PMID- 11010828 TI - An analysis of some hypotheses related to the Chingelput bacille Calmette-Guerin trial. AB - Investigation of several hypotheses related to the outcome of the Chingelput Bacille Calmette-Guerin (BCG) Trial suggests at least 2 factors that might explain the major scientific puzzle of a protective effect expected of 80% and a protective effect observed of 0% (i.e., equivalent protection for BCG and placebo). One factor that explains some of the low efficacy observed for BCG in this trial is the virtual saturation level of exposure to environmental mycobacteria (EM). Studies in animal models demonstrated that the protection afforded by infection with EM was equivalent to the protection that resulted from BCG vaccination. The second factor, pathogenetic pathway, explains why there was still a high case rate for tuberculosis, even though the population was fully vaccinated by EM. This hypothesis states that tuberculosis in India, as well as in most developing countries, results primarily from exogenous reinfection, a pathway against which BCG (or EM) exerts no protective effect beyond that induced by the first episode of infection with Mycobacterium tuberculosis. PMID- 11010829 TI - The Bcg host-resistance gene. AB - In the mouse, resistance and susceptibility to intracellular growth of mycobacteria in macrophages is controlled by the Bcg (Nramp1) gene, which has been cloned and shown to encode a macrophage phagosomal membrane protein with a putative transporter function. In the homologous human NRAMP1 gene, a total of 11 polymorphisms have been identified, which are being used to test for the linkage of NRAMP1 alleles with human responses to mycobacteria, including susceptibility to tuberculosis and leprosy, as well as BCG immunotherapy in bladder cancer. PMID- 11010830 TI - Efficacy and safety of bacille Calmette-Guerin immunotherapy in superficial bladder cancer. AB - In the United States, bladder cancer is the fourth most common human malignancy. In the past decade, the incidence of bladder cancer has increased by 36%. However, mortality has declined by 8%. Intravesical chemotherapy was considered to be partially responsible for this improvement in survival, but a recent review of clinical studies shows no reduction in disease progression with intravesical chemotherapy. Fortunately, the results of immunotherapy with bacille Calmette Guerin (BCG) are quite different, and it is expected that patients treated with optimal BCG treatment regimens will have a long-term reduction in tumor recurrence, tumor progression, and cancer mortality. PMID- 11010831 TI - Mechanisms of action of intravesical bacille Calmette-Guerin: local immune mechanisms. AB - The local immune response to mycobacteria is complex, but mycobacterial antigen presentation by phagocytes to T helper cells is the pivotal interaction. Bacille Calmette-Guerin (BCG) vaccination is associated with the development of antituberculosis immunity but not necessarily with antitumor immunity. Animal studies have shown that an intact host immune system is required for the antitumor activity of BCG. Immunosuppressed and, particularly, T cell-depleted individuals fail to respond to BCG immunotherapy. Clinical and laboratory evidence suggest that the antitumor activity is concentrated at the site of BCG administration, which reinforces the view that local immune mechanisms are responsible for this phenomenon. PMID- 11010832 TI - In vitro generation of bacillus Calmette-Guerin-activated killer cells. AB - Tumor regression induced in cancer patients by local instillation of bacillus Calmette-Guerin (BCG) into the bladder is considered to be mediated by cellular immune and inflammatory reactions. In an attempt to elucidate which of these effects are relevant to tumoricidal activity, an in vitro system was employed in which the immunostimulatory effects of BCG could be studied. This report describes the induction of BCG-activated killer (BAK) cells, which effectively lyse bladder tumor cells. Human peripheral blood mononuclear cells (PBMC) were stimulated with viable and sonicated BCG (v-BCG and s-BCG, respectively) to generate BAK cells. Cytotoxicity of BAK cells was comparable with the cytotoxicity exerted by lymphokine-activated killer (LAK) cells generated by interferon (IFN)-gamma but did not reach the level of interleukin-2 (IL-2) generated LAK cells. Induction of BAK cells was possible only with v-BCG and not with s-BCG. By depletion and enrichment of defined cell populations, the cytotoxic potential of BAK cells could be attributed to a population of CD8(+) and CD56(+) double-positive lymphocytes. Macrophages and CD4(+) cells were required for the induction of killing activity but had no such activity by themselves. Furthermore, the presence of IFN-gamma and IL-2 in the supernatants harvested during the generation of BAK cells was demonstrated. Monoclonal antibodies neutralizing these cytokines abolished BCG-mediated cytotoxicity. From these results, it is concluded that the known beneficial effect of local instillation of BCG on maintenance of the relapse-free state in superficial bladder cancer may be due to local generation of BAK cells. PMID- 11010833 TI - Reduction of side effects of intravesical therapy with bacille Calmette-Guerin by pentoxifylline?--an in vitro approach. AB - Immunotherapy with intravesical bacille Calmette-Guerin (BCG) is the treatment of choice against superficial bladder cancer recurrences. However, this therapy is associated with side effects that are considered to be the result of inflammatory cytokines. Since pentoxifylline is known to interfere with the production of cytokines, this drug was tested in vitro with regard to a later clinical application in BCG-treated patients. The cytokine release and the cytotoxicity of interleukin-2 or BCG-stimulated mononuclear cells were analyzed, and the growth of BCG under the influence of pentoxifylline was assayed. The results showed an inhibition of cytokine release of stimulated mononuclear cells. The cytotoxicity of BCG-stimulated mononuclear cells but not of lymphokine-stimulated mononuclear cells against bladder carcinoma cells was significantly inhibited. Restimulation with fresh BCG restored cytotoxicity. Direct coincubation of BCG and pentoxifylline resulted in a reduction of mycobacterial metabolism. From these data, we conclude that the use of pentoxifylline to reduce BCG-related side effects should be tested further in a clinical study. PMID- 11010834 TI - Role of animal models in understanding intravesical therapy with bacille Calmette Guerin. AB - Animal models provide a vehicle for understanding basic biological questions. Through the use of animal models, adequate control of experimental design is possible so that rigorous experiments can be performed to test a hypothesis. In the case of testing therapeutic mechanisms, it is important to select a model that is most analogous to the clinical setting so that observations can be readily transferred to clinical studies for validation. In mechanistic studies of Mycobacterium bovis strain (bacille Calmette-Guerin) therapy for bladder cancer, the orthotopic animal model most closely mimics the clinical treatment of bladder cancer. PMID- 11010835 TI - Immunostimulation in the urinary bladder by local application of Nocardia rubra cell-wall skeletons (Rubratin) and bacillus Calmette-Guerin as therapy for superficial bladder cancer: a comparative study. AB - Twelve patients with superficial bladder cancer were treated with intravesical instillations of Rubratin (ASTA Pharma AG, Frankfurt, Germany), a cell-wall preparation of Nocardia rubra. The objective was to compare the immunostimulating effect of Rubratin with that of bacillus Calmette-Guerin (BCG). Local immunostimulation was determined by cytokine induction in serially collected urine samples during the first 24 h after each instillation, leukocyte influx into the urine, and phenotypic analysis of the lymphocyte fraction. Levels of Rubratin-induced interleukin (IL)-1 beta, IL-6, and tumor necrosis factor-alpha were significantly elevated compared with pretherapy levels. Rubratin induced leukocyte influx into the urine. T-cell activation (IL-2 receptor and human leukocyte antigen-DR expression) can be induced, and CD4:CD8 cell ratios can be increased. All parameters indicated that Rubratin-induced immunostimulation was less than that associated with BCG. In conclusion, although local Rubratin induced immunostimulation occurs in a limited number of patients, the amount of immunocompetent cells attracted to the bladder seems to be less than that associated with BCG therapy, thus resulting in lower levels of cytokine production (which may reflect less clinical efficacy). PMID- 11010836 TI - Bacille Calmette-Guerin vaccine. PMID- 11010837 TI - HLA class I alleles associated with susceptibility or resistance to human immunodeficiency virus type 1 infection among a population in Chaco Province, Argentina. AB - Host genetic factors, such as HLA alleles, are important in human immunodeficiency virus (HIV) infection and its progression to AIDS. HLA class I gene products are involved in peptide presentation, and each allele is responsible for presenting a different set of peptides to cytotoxic T lymphocytes. The increase or decrease in the frequency of certain alleles in HIV 1-positive versus control subjects would suggest that alleles play a role in susceptibility to or protection against this viral infection. In the present study, 56 HIV-1-positive patients in Chaco Province, Argentina, were typed for the HLA-A and HLA-B loci, using polymerase chain reaction and sequence-specific oligonucleotide probes. The frequency of alleles A*24, B*18, and B*39 was increased in HIV-1-positive subjects, suggesting that these alleles play a role in susceptibility to HIV-1 infection. Alleles B*44 and B*55 were not found in HIV 1-positive subjects, suggesting that they have a protective effect against the disease. The P values of the alleles B*39 and B*44 were statistically significant (P<.05). PMID- 11010838 TI - Evaluation of a live, cold-passaged, temperature-sensitive, respiratory syncytial virus vaccine candidate in infancy. AB - A live-attenuated, intranasal respiratory syncytial virus (RSV) candidate vaccine, cpts-248/404, was tested in phase 1 trials in 114 children, including 37 1-2-month-old infants-a target age for RSV vaccines. The cpts-248/404 vaccine was infectious at 104 and 105 plaque-forming units in RSV-naive children and was broadly immunogenic in children >6 months old. Serum and nasal antibody responses in 1-2 month olds were restricted to IgA, had a dominant response to RSV G protein, and had no increase in neutralizing activity. Nevertheless, there was restricted virus shedding on challenge with a second vaccine dose and preliminary evidence for protection from symptomatic disease on natural reexposure. The cpts 248/404 vaccine candidate did not cause fever or lower respiratory tract illness. In the youngest infants, however, cpts-248/404 was unacceptable because of upper respiratory tract congestion associated with peak virus recovery. A live attenuated RSV vaccine for the youngest infant will use cpts-248/404 modified by additional attenuating mutations. PMID- 11010839 TI - Treatment-mediated changes in human immunodeficiency virus (HIV) type 1 RNA and CD4 cell counts as predictors of weight growth failure, cognitive decline, and survival in HIV-infected children. AB - This meta-analysis of 5 large studies of the Pediatric AIDS Clinical Trials Group was undertaken to evaluate the predictive value of antiretroviral treatment mediated changes in 3 markers of human immunodeficiency virus (HIV) type 1 disease progression-HIV-1 RNA level, CD4 cell count, and CD4 percentage-for weight growth failure, cognitive decline, and survival in HIV-infected children. Proportional hazards models were used to assess the prognostic value of the markers at baseline and after 24 weeks of treatment, with data from 1089 children. Among children receiving nucleoside with or without nonnucleoside reverse-transcriptase inhibitors, higher immunologic and lower virologic markers at baseline and after 24 weeks were significant independent predictors of survival, whereas virologic markers were significant predictors of weight growth and cognitive failure in children >1 year old. The finding of differential age effects on pediatric-specific clinical outcomes emphasizes the need for continued investigation of treatment effects in children. PMID- 11010840 TI - The relationship between virus load response to highly active antiretroviral therapy and change in CD4 cell counts: A report from the Women's interagency HIV study. AB - The relationship between the pattern of virus load response to highly active antiretroviral therapy (HAART) and CD4 lymphocyte response was assessed in a cohort of 249 human immunodeficiency virus (HIV) type 1-infected women at 3 times: 1 before and 2 after initiation of therapy, with follow-up of 6-12 months. Patients with a durable response to HAART (i.e., >1 log decrease in HIV-1 RNA sustained for the study periods) had a continuous and significant increase in CD4 cell counts over time, whereas those with no response (<0.5 log decrease in HIV-1 RNA) had a slight decline. Patients with a mixed response (initial decrease >1 log, followed by a subsequent decrease <0.5 log) had an increase in CD4 cell count, followed by a plateau. The trend in CD4 cell count differed significantly by response to HAART, with those patients who experienced a durable response having significantly higher CD4 cell counts than others. PMID- 11010841 TI - Infectious disease morbidity in the US region bordering Mexico, 1990-1998. AB - The United States and Mexico share an international boundary approximately 3000 km long. This border separates 2 nations with great differences in health status. The objective of this study was to assess morbidity due to infectious diseases in the US region bordering Mexico. The incidence between 1990 and 1998 of 22 nationally notifiable infectious diseases was compared between border and nonborder regions. Disease rates, reflected as rate ratios, were higher in the border region for botulism, brucellosis, diphtheria, hepatitis A, measles, mumps, rabies, rubella, salmonellosis, and shigellosis than in either of 2 nonborder comparison regions. These data indicate that incidence rates for a variety of infectious diseases of public health importance are significantly higher in the United States along the Mexican border than in nonborder regions. These results suggest that an inadequate public health infrastructure may contribute to excess morbidity due to infectious diseases in the border region. PMID- 11010842 TI - Phylogenetic subgroups of human T cell lymphotropic virus (HTLV) type I in the tax gene and their association with different risks for HTLV-I-associated myelopathy/tropical spastic paraparesis. AB - The association between human T cell lymphotropic virus (HTLV) type I tax variation and disease outcome was studied. The tax gene was sequenced in 61 patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), 55 patients with adult T cell leukemia, and 62 healthy carriers (HCs). Phylogenetic analysis revealed 2 tax gene subgroups that are related on the basis of the long terminal repeat sequence. Further analysis using restriction fragment length polymorphism in 192 patients with HAM/TSP and 200 HCs revealed a higher incidence of 1 tax subgroup (taxA) in HAM/TSP. taxA was present in 30 (15.5%) of 192 patients with HAM/TSP and in 14 (7%) of 200 HCs. The difference was significant (chi2=6.47; P=.014; odds ratio, 2.46; 95% confidence interval, 1.26 4.80). This effect was independent of HLA-A*02, which has been reported to prevent HAM/TSP development. These findings suggest that both host genetic factors and HTLV-I subgroup are associated with different risks for development of HAM/TSP. PMID- 11010856 TI - Inactivation of Escherichia coli and Listeria innocua in milk by combined treatment with high hydrostatic pressure and the lactoperoxidase system. AB - We have studied inactivation of four strains each of Escherichia coli and Listeria innocua in milk by the combined use of high hydrostatic pressure and the lactoperoxidase-thiocyanate-hydrogen peroxide system as a potential mild food preservation method. The lactoperoxidase system alone exerted a bacteriostatic effect on both species for at least 24 h at room temperature, but none of the strains was inactivated. Upon high-pressure treatment in the presence of the lactoperoxidase system, different results were obtained for E. coli and L. innocua. For none of the E. coli strains did the lactoperoxidase system increase the inactivation compared to a treatment with high pressure alone. However, a strong synergistic interaction of both treatments was observed for L. innocua. Inactivation exceeding 7 decades was achieved for all strains with a mild treatment (400 MPa, 15 min, 20 degrees C), which in the absence of the lactoperoxidase system caused only 2 to 5 decades of inactivation depending on the strain. Milk as a substrate was found to have a considerable effect protecting E. coli and L. innocua against pressure inactivation and reducing the effectiveness of the lactoperoxidase system under pressure on L. innocua. Time course experiments showed that L. innocua counts continued to decrease in the first hours after pressure treatment in the presence of the lactoperoxidase system. E. coli counts remained constant for at least 24 h, except after treatment at the highest pressure level (600 MPa, 15 min, 20 degrees C), in which case, in the presence of the lactoperoxidase system, a transient decrease was observed, indicating sublethal injury rather than true inactivation. PMID- 11010857 TI - Differential C isotope discrimination by fungi during decomposition of C(3)- and C(4)-derived sucrose. AB - Stable isotope analysis is a major tool used in ecosystem studies to establish pathways and rates of C exchange between various ecosystem components. Little is known about isotopic effects of many such components, especially microbes. Here we report on the discovery of an unexpected pattern of C isotopic discrimination by basidiomycete fungi with far-reaching consequences for our understanding of isotopic processing in ecosystems where these microbes mediate material transfers across trophic levels. We measured fractionation effects on three ecologically relevant basidiomycete species under controlled laboratory conditions. Sucrose derived from C(3) and C(4) plants is fractionated differentially by these microbes in a taxon-specific manner. The differentiation between mycorrhizal and saprotrophic fungi observed in the field by others is not explained by intrinsic discrimination patterns. Fractionation occurs during sugar uptake and is sensitive to the nonrandom distribution of stable isotopes in the sucrose molecule. The balance between respiratory physiology and fermentative physiology modulates the degree of fractionation. These discoveries disprove the assumption that fungal C processing does not significantly alter the distribution of stable C isotopes and provide the basis for a reevaluation of ecosystem models based on isotopic evidence that involve C transfer across microbial interfaces. We provide a mechanism to account for the observed differential discrimination effects. PMID- 11010858 TI - Urea hydrogen peroxide reduces the numbers of lactobacilli, nourishes yeast, and leaves no residues in the ethanol fermentation. AB - Urea hydrogen peroxide (UHP) at a concentration of 30 to 32 mmol/liter reduced the numbers of five Lactobacillus spp. (Lactobacillus plantarum, L. paracasei, Lactobacillus sp. strain 3, L. rhamnosus, and L. fermentum) from approximately 10(7) to approximately 10(2) CFU/ml in a 2-h preincubation at 30 degrees C of normal-gravity wheat mash at approximately 21 g of dissolved solids per ml containing normal levels of suspended grain particles. Fermentation was completed 36 h after inoculation of Saccharomyces cerevisiae in the presence of UHP, even when wheat mash was deliberately contaminated (infected) with L. paracasei at approximately 10(7) CFU/ml. There were no significant differences in the maximum ethanol produced between treatments when urea hydrogen peroxide was used to kill the bacteria and controls (in which no bacteria were added). However, the presence of L. paracasei at approximately 10(7) CFU/ml without added agent resulted in a 5.84% reduction in the maximum ethanol produced compared to the control. The bactericidal activity of UHP is greatly affected by the presence of particulate matter. In fact, only 2 mmol of urea hydrogen peroxide per liter was required for disinfection when mashes had little or no particulate matter present. No significant differences were observed in the decomposition of hydrogen peroxide in normal-gravity wheat mash at 30 degrees C whether the bactericidal agent was added as H(2)O(2) or as urea hydrogen peroxide. NADH peroxidase activity (involved in degrading H(2)O(2)) increased significantly (P = 0.05) in the presence of 0.75 mM hydrogen peroxide (sublethal level) in all five strains of lactobacilli tested but did not persist in cells regrown in the absence of H(2)O(2). H(2)O(2)-resistant mutants were not expected or found when lethal levels of H(2)O(2) or UHP were used. Contaminating lactobacilli can be effectively managed by UHP, a compound which when used at ca. 30 mmol/liter happens to provide near-optimum levels of assimilable nitrogen and oxygen that aid in vigorous fermentation performance by yeast. PMID- 11010859 TI - Seasonal population dynamics and interactions of competing bacteriophages and their host in the rhizosphere. AB - We describe two prolonged bacteriophage blooms within sugar beet rhizospheres ensuing from an artificial increase in numbers of an indigenous soil bacterium. Further, we provide evidence of in situ competition between these phages. This is the first in situ demonstration of such microbial interactions in soil. To achieve this, sugar beet seeds were inoculated with Serratia liquefaciens CP6RS or its lysogen, CP6RS-ly-phi 1. These were sown, along with uninoculated seeds, in 36 field plots arranged in a randomized Latin square. The plots were then sampled regularly over 194 days, and the plants were assayed for the released bacteria and any infectious phages. Both the lysogen and nonlysogen forms of CP6RS survived equally well in situ, contradicting earlier work suggesting lysogens have a competitive disadvantage in nature. A Podoviridae phage, identified as phi CP6-4, flourished on the nonlysogen-inoculated plants in contrast to those plants inoculated with the lysogen. Conversely, the Siphoviridae phage phi CP6-1 (used to construct the released lysogen) was isolated abundantly from the lysogen-treated plants but almost never on the nonlysogen-inoculated plants. The uninoculated plants also harbored some phi CP6 1 phage up to day 137, yet hardly any phi CP6-4 phages were found, and this was consistent with previous years. We show that the different temporal and spatial distributions of these two physiologically distinct phages can be explained by application of optimal foraging theory to phage ecology. This is the first time that such in situ evidence has been provided in support of this theoretical model. PMID- 11010860 TI - A strain of Enterococcus faecium (18C23) inhibits adhesion of enterotoxigenic Escherichia coli K88 to porcine small intestine mucus. AB - Few studies, if any, have addressed the adhesion of enterococci to the intestinal mucosa and their interference with the adhesion of pathogens, although more than 60% of probiotic preparations in the market contain strains of enterococci. The objective of this study was to investigate if Enterococcus faecium 18C23 has the ability to inhibit the adhesion of Escherichia coli K88ac and K88MB to the small intestine mucus of piglets. Approximately 9% of E. faecium 18C23 organisms adhered to the small intestine mucus, and the adhesion was found to be specific. Living E. faecium 18C23 culture efficiently inhibited the adhesion of E. coli K88ac and K88MB to the piglet intestine mucus. Inhibition of the adhesion of E. coli K88ac to the small intestine mucus was found to be dose dependent. Inhibition of >90% was observed when 10(9) CFU or more of living E. faecium 18C23 culture per ml was added simultaneously with E. coli to immobilized mucus. The substances from both the 18C23 cells and the spent culture supernatant contributed to the inhibition of adhesion of E. coli K88 to the small intestine mucus receptors. The inhibiting effect was not solely a pH effect since considerable inhibitory action was demonstrated after neutralizing the mixture or spent culture supernatant to pH 7.0. Part of the inhibition of adhesion of E. coli K88ac by E. faecium 18C23 or its supernatant might occur through steric hindrance. PMID- 11010861 TI - Rapid mineralization of benzo[a]pyrene by a microbial consortium growing on diesel fuel. AB - A microbial consortium which rapidly mineralized the environmentally persistent pollutant benzo[a]pyrene was recovered from soil. The consortium cometabolically converted [7-(14)C]benzo[a]pyrene to (14)CO(2) when it was grown on diesel fuel, and the extent of benzo[a]pyrene mineralization was dependent on both diesel fuel and benzo[a]pyrene concentrations. Addition of diesel fuel at concentrations ranging from 0.007 to 0.2% (wt/vol) stimulated the mineralization of 10 mg of benzo[a]pyrene per liter 33 to 65% during a 2-week incubation period. When the benzo[a]pyrene concentration was 10 to 100 mg liter(-1) and the diesel fuel concentration was 0.1% (wt/vol), an inoculum containing 1 mg of cell protein per liter (small inoculum) resulted in mineralization of up to 17.2 mg of benzo[a]pyrene per liter in 16 days. This corresponded to 35% of the added radiolabel when the concentration of benzo[a]pyrene was 50 mg liter(-1). A radiocarbon mass balance analysis recovered 25% of the added benzo[a]pyrene solubilized in the culture suspension prior to mineralization. Populations growing on diesel fuel most likely promoted emulsification of benzo[a]pyrene through the production of surface-active compounds. The consortium was also analyzed by PCR-denaturing gradient gel electrophoresis of 16S rRNA gene fragments, and 12 dominant bands, representing different sequence types, were detected during a 19-day incubation period. The onset of benzo[a]pyrene mineralization was compared to changes in the consortium community structure and was found to correlate with the emergence of at least four sequence types. DNA from 10 sequence types were successfully purified and sequenced, and that data revealed that eight of the consortium members were related to the class Proteobacteria but that the consortium also included members which were related to the genera Mycobacterium and Sphingobacterium. PMID- 11010862 TI - Chemostat enrichments of human feces with resistant starch are selective for adherent butyrate-producing clostridia at high dilution rates. AB - Resistant starch (RS) enrichments were made using chemostats inoculated with human feces from two individuals at two dilution rates (D = 0.03 h(-1) and D = 0.30 h(-1)) to select for slow- and fast-growing amylolytic communities. The fermentations were studied by analysis of short-chain fatty acids, amylase and alpha-glucosidase activities, and viable counts of the predominant culturable populations and the use of 16S rRNA-targeted oligonucleotide probes. Considerable butyrate was produced at D = 0. 30 h(-1), which corresponded with reduced branched-chain fatty acid formation. At both dilution rates, high levels of extracellular amylase activity were produced, while alpha-glucosidase was predominantly cell associated. Bacteroides and bifidobacteria predominated at the low dilution rate, whereas saccharolytic clostridia became more important at D = 0.30 h(-1). Microscopic examination showed that within 48 h of inoculation, one particular bacterial morphotype predominated in RS enrichments at D = 0.30 h(-1). This organism attached apically to RS granules and formed rosette-like structures which, with glycocalyx formation, agglomerated to form biofilm networks in the planktonic phase. Attempts to isolate this bacterium in pure culture were repeatedly unsuccessful, although a single colony was eventually obtained. On the basis of its 16S rDNA sequence, this RS-degrading, butyrate-producing organism was identified as being a previously unidentified group I Clostridium sp. A 16S rRNA-targeted probe was designed using this sequence and used to assess the abundance of the population in the enrichments. At 240 h, its contributions to total rRNA in the chemostats were 5 and 23% at D = 0.03 and 0.30 h(-1), respectively. This study indicates that bacterial populations with significant metabolic potential can be overlooked using culture-based methodologies. This may provide a paradigm for explaining the discrepancy between the low numbers of butyrate-producing bacteria that are isolated from fecal samples and the actual production of butyrate. PMID- 11010863 TI - Evolution of thermotolerance in hot spring cyanobacteria of the genus Synechococcus. AB - The extension of ecological tolerance limits may be an important mechanism by which microorganisms adapt to novel environments, but it may come at the evolutionary cost of reduced performance under ancestral conditions. We combined a comparative physiological approach with phylogenetic analyses to study the evolution of thermotolerance in hot spring cyanobacteria of the genus Synechococcus. Among the 20 laboratory clones of Synechococcus isolated from collections made along an Oregon hot spring thermal gradient, four different 16S rRNA gene sequences were identified. Phylogenies constructed by using the sequence data indicated that the clones were polyphyletic but that three of the four sequence groups formed a clade. Differences in thermotolerance were observed for clones with different 16S rRNA gene sequences, and comparison of these physiological differences within a phylogenetic framework provided evidence that more thermotolerant lineages of Synechococcus evolved from less thermotolerant ancestors. The extension of the thermal limit in these bacteria was correlated with a reduction in the breadth of the temperature range for growth, which provides evidence that enhanced thermotolerance has come at the evolutionary cost of increased thermal specialization. This study illustrates the utility of using phylogenetic comparative methods to investigate how evolutionary processes have shaped historical patterns of ecological diversification in microorganisms. PMID- 11010864 TI - Biochemical and genetic characterization of propionicin T1, a new bacteriocin from Propionibacterium thoenii. AB - A collection of propionibacteria was screened for bacteriocin production. A new bacteriocin named propionicin T1 was isolated from two strains of Propionibacterium thoenii. This bacteriocin shows no sequence similarity to other bacteriocins. Propionicin T1 was active against all strains of Propionibacterium acidipropionici, Propionibacterium thoenii, and Propionibacterium jensenii tested and also against Lactobacillus sake NCDO 2714 but showed no activity against Propionibacterium freudenreichii. The bacteriocin was purified, and the N terminal part of the peptide was determined with amino acid sequencing. The corresponding gene pctA was sequenced, and this revealed that propionicin T1 is produced as a prebacteriocin of 96 amino acids with a typical sec leader, which is processed to give a mature bacteriocin of 65 amino acids. An open reading frame encoding a protein of 424 amino acids was found 68 nucleotides downstream the stop codon of pctA. The N-terminal part of this putative protein shows strong similarity with the ATP-binding cassette of prokaryotic and eukaryotic ABC transporters, and this protein may be involved in self-protection against propionicin T1. Propionicin T1 is the first bacteriocin from propionibacteria that has been isolated and further characterized at the molecular level. PMID- 11010865 TI - Bacterial community structure associated with a dimethylsulfoniopropionate producing North Atlantic algal bloom. AB - The bacteria associated with oceanic algal blooms are acknowledged to play important roles in carbon, nitrogen, and sulfur cycling, yet little information is available on their identities or phylogenetic affiliations. Three culture independent methods were used to characterize bacteria from a dimethylsulfoniopropionate (DMSP)-producing algal bloom in the North Atlantic. Group-specific 16S rRNA-targeted oligonucleotides, 16S ribosomal DNA (rDNA) clone libraries, and terminal restriction fragment length polymorphism analysis all indicated that the marine Roseobacter lineage was numerically important in the heterotrophic bacterial community, averaging >20% of the 16S rDNA sampled. Two other groups of heterotrophic bacteria, the SAR86 and SAR11 clades, were also shown by the three 16S rRNA-based methods to be abundant in the bloom community. In surface waters, the Roseobacter, SAR86, and SAR11 lineages together accounted for over 50% of the bacterial rDNA and showed little spatial variability in abundance despite variations in the dominant algal species. Depth profiles indicated that Roseobacter phylotype abundance decreased with depth and was positively correlated with chlorophyll a, DMSP, and total organic sulfur (dimethyl sulfide plus DMSP plus dimethyl sulfoxide) concentrations. Based on these data and previous physiological studies of cultured Roseobacter strains, we hypothesize that this lineage plays a role in cycling organic sulfur compounds produced within the bloom. Three other abundant bacterial phylotypes (representing a cyanobacterium and two members of the alpha Proteobacteria) were primarily associated with chlorophyll-rich surface waters of the bloom (0 to 50 m), while two others (representing Cytophagales and delta Proteobacteria) were primarily found in deeper waters (200 to 500 m). PMID- 11010866 TI - Substrate specificity of atrazine chlorohydrolase and atrazine-catabolizing bacteria. AB - Bacterial atrazine catabolism is initiated by the enzyme atrazine chlorohydrolase (AtzA) in Pseudomonas sp. strain ADP. Other triazine herbicides are metabolized by bacteria, but the enzymological basis of this is unclear. Here we begin to address this by investigating the catalytic activity of AtzA by using substrate analogs. Purified AtzA from Pseudomonas sp. strain ADP catalyzed the hydrolysis of an atrazine analog that was substituted at the chlorine substituent by fluorine. AtzA did not catalyze the hydrolysis of atrazine analogs containing the pseudohalide azido, methoxy, and cyano groups or thiomethyl and amino groups. Atrazine analogs with a chlorine substituent at carbon 2 and N-alkyl groups, ranging in size from methyl to t-butyl, all underwent dechlorination by AtzA. AtzA catalyzed hydrolytic dechlorination when one nitrogen substituent was alkylated and the other was a free amino group. However, when both amino groups were unalkylated, no reaction occurred. Cell extracts were prepared from five strains capable of atrazine dechlorination and known to contain atzA or closely homologous gene sequences: Pseudomonas sp. strain ADP, Rhizobium strain PATR, Alcaligenes strain SG1, Agrobacterium radiobacter J14a, and Ralstonia picketti D. All showed identical substrate specificity to purified AtzA from Pseudomonas sp. strain ADP. Cell extracts from Clavibacter michiganensis ATZ1, which also contains a gene homologous to atzA, were able to transform atrazine analogs containing pseudohalide and thiomethyl groups, in addition to the substrates used by AtzA from Pseudomonas sp. strain ADP. This suggests that either (i) another enzyme(s) is present which confers the broader substrate range or (ii) the AtzA itself has a broader substrate range. PMID- 11010867 TI - A methylotrophic pathway participates in pectin utilization by Candida boidinii. AB - The methylotrophic yeast Candida boidinii S2 was found to be able to grow on pectin or polygalacturonate as a carbon source. When cells were grown on 1% (wt/vol) pectin, C. boidinii exhibited induced levels of the pectin depolymerizing enzymes pectin methylesterase (208 mU/mg of protein), pectin lyase (673 mU/mg), pectate lyase (673 mU/mg), and polygalacturonase (3.45 U/mg) and two methanol-metabolizing peroxisomal enzymes, alcohol oxidase (0.26 U/mg) and dihydroxyacetone synthase (94 mU/mg). The numbers of peroxisomes also increased ca. two- to threefold in cells grown on these pectic compounds (3.34 and 2.76 peroxisomes/cell for cells grown on pectin and polygalacturonate, respectively) compared to the numbers in cells grown on glucose (1.29 peroxisomes/cell). The cell density obtained with pectin increased as the degree of methyl esterification of pectic compounds increased, and it decreased in strains from which genes encoding alcohol oxidase and dihydroxyacetone synthase were deleted and in a peroxisome assembly mutant. Our study showed that methanol metabolism and peroxisome assembly play important roles in the degradation of pectin, especially in the utilization of its methyl ester moieties. PMID- 11010868 TI - A bead-based method for multiplexed identification and quantitation of DNA sequences using flow cytometry. AB - A new multiplexed, bead-based method which utilizes nucleic acid hybridizations on the surface of microscopic polystyrene spheres to identify specific sequences in heterogeneous mixtures of DNA sequences is described. The method consists of three elements: beads (5.6-microm diameter) with oligomer capture probes attached to the surface, three fluorophores for multiplexed detection, and flow cytometry instrumentation. Two fluorophores are impregnated within each bead in varying amounts to create different bead types, each associated with a unique probe. The third fluorophore is a reporter. Following capture of fluorescent cDNA sequences from environmental samples, the beads are analyzed by flow cytometric techniques which yield a signal intensity for each capture probe proportional to the amount of target sequences in the analyte. In this study, a direct hybrid capture assay was developed and evaluated with regard to sequence discrimination and quantitation of abundances. The target sequences (628 to 728 bp in length) were obtained from the 16S/23S intergenic spacer region of microorganisms collected from polluted groundwater at the nuclear waste site in Hanford, Wash. A fluorescence standard consisting of beads with a known number of fluorescent DNA molecules on the surface was developed, and the resolution, sensitivity, and lower detection limit for measuring abundances were determined. The results were compared with those of a DNA microarray using the same sequences. The bead method exhibited far superior sequence discrimination and possesses features which facilitate accurate quantitation. PMID- 11010869 TI - Application of 5'-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk. AB - PCR techniques have significantly improved the detection and identification of bacterial pathogens. Countless adaptations and applications have been described, including quantitative PCR and the latest innovation, real-time PCR. In real-time PCR, e.g., the 5'-nuclease chemistry renders the automated and direct detection and quantification of PCR products possible (P. M. Holland et al., Proc. Natl. Acad. Sci. USA 88:7276-7280, 1991). We present an assay for the quantitative detection of Listeria monocytogenes based on the 5'-nuclease PCR using a 113-bp amplicon from the listeriolysin O gene (hlyA) as the target. The assay was positive for all isolates of L. monocytogenes tested (65 isolates including the type strain) and negative for all other Listeria strains (16 isolates from five species tested) and several other bacteria (18 species tested). The application of 5'-nuclease PCR in diagnostics requires a quantitative sample preparation step. Several magnetic bead-based strategies were evaluated, since these systems are simple and relatively easy to automate. The combination of nonspecific binding of bacteria to paramagnetic beads, with subsequent DNA purification by use of the same beads, gave the most satisfactory result. The detection limit was approximately 6 to 60 CFU, quantification was linear over at least 7 log units, and the method could be completed within 3 h. In conclusion, a complete quantitative method for L. monocytogenes in water and in skimmed and raw milk was developed. PMID- 11010870 TI - Development of genetic tools for Lactobacillus sakei: disruption of the beta galactosidase gene and use of lacZ as a reporter gene To study regulation of the putative copper ATPase, AtkB. AB - Downstream from the ptsHI operon of Lactobacillus sakei, the genes atkY and atkB, organized in an operon, were observed. The two putative proteins, AtkB and AtkY, show sequence similarity to the Enterococcus hirae copper P-type ATPase, responsible for copper efflux, and its negative regulator. Characterization of AtkB as a copper P-type ATPase could not be demonstrated since an atkB mutant did not show any phenotype. Thus, another strategy was followed in order to investigate the transcriptional regulation of the atkYB locus, leading to the development of new genetic tools for L. sakei. A plasmid was constructed, the use of which allowed gene replacement at the lacLM locus in L. sakei by two successive crossovers. A strain deleted of the lacLM operon encoding the beta galactosidase of L. sakei was constructed by this method, and the Escherichia coli lacZ gene could then be used as a reporter gene to investigate the regulation of atkYB. Results show that the atkYB operon is induced by small concentrations of CuSO(4) (30 to 40 microM) but not when CuSO(4) is omitted or added at higher concentrations. PMID- 11010871 TI - Characterization, seasonal occurrence, and diel fluctuation of poly(hydroxyalkanoate) in photosynthetic microbial mats. AB - In situ poly(hydroxyalkanoate) (PHA) levels and repeating-unit compositions were examined in stratified photosynthetic microbial mats from Great Sippewissett Salt Marsh, Mass., and Ebro Delta, Spain. Unlike what has been observed in pure cultures of phototrophic bacteria, the prevalence of hydroxyvalerate (HV) repeating units relative to hydroxybutyrate (HB) repeating units was striking. In the cyanobacteria-dominated green material of Sippewissett mats, the mole percent ratio of repeating units was generally 1HB:1HV. In the purple sulfur bacteria dominated pink material the relationship was typically 1HB:2HV. In Sippewissett mats, PHA contributed about 0.5 to 1% of the organic carbon in the green layer and up to 6% in the pink layer. In Ebro Delta mats, PHA of approximately 1HB:2HV repeating-unit distribution contributed about 2% of the organic carbon of the composite photosynthetic layers (the green and pink layers were not separated). Great Sippewissett Salt Marsh mats were utilized for more extensive investigation of seasonal, diel, and exogenous carbon effects. When the total PHA content was normalized to organic carbon, there was little seasonal variation in PHA levels. However, routine daily variation was evident at all sites and seasons. In every case, PHA levels increased during the night and decreased during the day. This phenomenon was conspicuous in the pink layer, where PHA levels doubled overnight. The daytime declines could be inhibited by artificial shading. Addition of exogenous acetate, lactate, and propionate induced two- to fivefold increases in the total PHA levels when applied in the daylight but had no effect when applied at night. The distinct diel pattern of in situ PHA accumulation at night appears to be related, in some phototrophs, to routine dark energy metabolism and is not influenced by the availability of organic nutrients. PMID- 11010872 TI - Generation of new hydrogen-recycling Rhizobiaceae strains by introduction of a novel hup minitransposon. AB - Hydrogen evolution by nitrogenase is a source of inefficiency for the nitrogen fixation process by the Rhizobium-legume symbiosis. To develop a strategy to generate rhizobial strains with H(2)-recycling ability, we have constructed a Tn5 derivative minitransposon (TnHB100) that contains the ca. 18-kb H(2) uptake (hup) gene cluster from Rhizobium leguminosarum bv. viciae UPM791. Bacteroids from TnHB100-containing strains of R. leguminosarum bv. viciae PRE, Bradyrhizobium japonicum, R. etli, and Mesorhizobium loti expressed high levels of hydrogenase activity that resulted in full recycling of the hydrogen evolved by nitrogenase in nodules. Efficient processing of the hydrogenase large subunit (HupL) in these strains was shown by immunoblot analysis of bacteroid extracts. In contrast, Sinorhizobium meliloti, M. ciceri, and R. leguminosarum bv. viciae UML2 strains showed poor expression of the hup system that resulted in H(2)-evolving nodules. For the latter group of strains, no immunoreactive material was detected in bacteroid extracts using anti-HupL antiserum, suggesting a low level of transcription of hup genes or HupL instability. A general procedure for the characterization of the minitransposon insertion site and removal of antibiotic resistance gene included in TnHB100 has been developed and used to generate engineered strains suitable for field release. PMID- 11010873 TI - Cloning, expression, and characterization of the katG gene, encoding catalase peroxidase, from the polycyclic aromatic hydrocarbon-degrading bacterium Mycobacterium sp. strain PYR-1. AB - A 81-kDa protein from Mycobacterium sp. strain PYR-1 was expressed in response to exposure of the strain to the polycyclic aromatic hydrocarbon pyrene and recovered by two-dimensional gel electrophoresis. The N-terminal sequence of the protein indicated that it was similar to catalase-peroxidase. An oligonucleotide probe designed from this sequence was used to screen a genomic library of Mycobacterium sp. strain PYR-1, and a positive clone, containing a part of the gene encoding the 81-kDa protein, was isolated. A gene-walking technique was used to sequence the entire gene, which was identified as katG for catalase peroxidase. The deduced KatG protein sequence showed significant homology to KatGII of Mycobacterium fortuitum and clustered with catalase-peroxidase proteins from other Mycobacterium species in a phylogenetic tree. The katG gene was expressed in Escherichia coli to produce a protein with catalase-peroxidase activity. Since the induction of this catalase-peroxidase occurred in pyrene induced cultures and the exposure of these cultures to hydrogen peroxide reduced pyrene metabolism, our data suggest that this enzyme plays a role in polycyclic aromatic hydrocarbon metabolism by strain PYR-1. PMID- 11010874 TI - Cytological effects of cellulases in the parasitism of Phytophthora parasitica by Pythium oligandrum. AB - The ubiquitous oomycete Pythium oligandrum is a potential biocontrol agent for use against a wide range of pathogenic fungi and an inducer of plant disease resistance. The ability of P. oligandrum to compete with root pathogens for saprophytic colonization of substrates may be critical for pathogen increase in soil, but other mechanisms, including antibiosis and enzyme production, also may play a role in the antagonistic process. We used transmission electron microscopy and gold cytochemistry to analyze the intercellular interaction between P. oligandrum and Phytophthora parasitica. Growth of P. oligandrum towards Phytophthora cells correlated with changes in the host, including retraction of the plasma membrane and cytoplasmic disorganization. These changes were associated with the deposition onto the inner host cell surface of a cellulose enriched material. P. oligandrum hyphae could penetrate the thickened host cell wall and the cellulose-enriched material, suggesting that large amounts of cellulolytic enzymes were produced. Labeling of cellulose with gold-complexed exoglucanase showed that the integrity of the cellulose was greatly affected both along the channel of fungal penetration and also at a distance from it. We measured cellulolytic activity of P. oligandrum in substrate-free liquid medium. The enzymes present were almost as effective as those from Trichoderma viride in degrading both carboxymethyl cellulose and Phytophthora wall-bound cellulose. P. oligandrum and its cellulolytic enzymes may be useful for biological control of oomycete pathogens, including Phytophthora and Pythium spp., which are frequently encountered in field and greenhouse production. PMID- 11010875 TI - Quantitative flow cytometric evaluation of maximal Cryptosporidium parvum oocyst infectivity in a neonate mouse model. AB - The importance of waterborne transmission of Cryptosporidium parvum to humans has been highlighted by recent outbreaks of cryptosporidiosis. The first step in a survey of contaminated water currently consists of counting C. parvum oocysts. Data suggest that an accurate risk evaluation should include a determination of viability and infectivity of counted oocysts in water. In this study, oocyst infectivity was addressed by using a suckling mouse model. Four-day-old NMRI (Naval Medical Research Institute) mice were inoculated per os with 1 to 1,000 oocysts in saline. Seven days later, the number of oocysts present in the entire small intestine was counted by flow cytometry using a fluorescent, oocyst specific monoclonal antibody. The number of intestinal oocysts was directly related to the number of inoculated oocysts. For each dose group, infectivity of oocysts, expressed as the percentage of infected animals, was 100% for challenge doses between 25 and 1,000 oocysts and about 70% for doses ranging from 1 to 10 oocysts/animal. Immunofluorescent flow cytometry was useful in enhancing the detection sensitivity in the highly susceptible NMRI suckling mouse model and so was determined to be suitable for the evaluation of maximal infectivity risk. PMID- 11010876 TI - Use of two-dimensional electrophoresis To study differential protein expression in divercin V41-resistant and wild-type strains of Listeria monocytogenes. AB - The use of bacteriocins from food-grade lactic acid bacteria to fight against the food-borne pathogen Listeria monocytogenes has been gaining interest. However, the emergence of resistant cells is frequently reported when Listeria is exposed to such antibacterials. A two-dimensional electrophoresis study of whole-cell protein expression of Listeria monocytogenes variants sensitive or resistant to the action of a bacteriocin produced by Carnobacterium divergens V41, divercin V41, is reported in this paper. The resistant variant obtained from the sensitive strain of L. monocytogenes P was also resistant to piscicocins V1 and SF668, but remained sensitive to nisin. Its growth rate was 50% less than the sensitive strain, and the MIC for it was 10(4) times higher. No reversion of the resistance was observed after 20 successive cultures in the absence of divercin V41. Comparison of the protein patterns by two-dimensional gel electrophoresis analysis showed clear differences. In the resistant variant pattern, at least nine spots had disappeared and eight new ones were observed. One of the newly synthesized proteins was identified as a flagellin of L. monocytogenes. Direct interaction between flagellin and divercin V41 was not evidenced. Intracellular synthesis of flagellin is probably an indirect effect of a modification in transcriptional regulation with widespread effects through a sigma factor. An intense protein, only present in the sensitive strain, was identified as a non heme iron-binding ferritin displaying strong similarities to Dps proteins. Common modifications in the transcriptional regulation for these two proteins are discussed. PMID- 11010877 TI - Characterization of reutericyclin produced by Lactobacillus reuteri LTH2584. AB - Lactobacillus reuteri LTH2584 exhibits antimicrobial activity that can be attributed neither to bacteriocins nor to the production of reuterin or organic acids. We have purified the active compound, named reutericyclin, to homogeneity and characterized its antimicrobial activity. Reutericyclin exhibited a broad inhibitory spectrum including Lactobacillus spp., Bacillus subtilis, B. cereus, Enterococcus faecalis, Staphylococcus aureus, and Listeria innocua. It did not affect the growth of gram-negative bacteria; however, the growth of lipopolysaccharide mutant strains of Escherichia coli was inhibited. Reutericyclin exhibited a bactericidal mode of action against Lactobacillus sanfranciscensis, Staphylococcus aureus, and B. subtilis and triggered the lysis of cells of L. sanfranciscensis in a dose-dependent manner. Germination of spores of B. subtilis was inhibited, but the spores remained unaffected under conditions that do not permit germination. The fatty acid supply of the growth media had a strong effect on reutericyclin production and its distribution between producer cells and the culture supernatant. Reutericyclin was purified from cell extracts and culture supernatant of L. reuteri LTH2584 cultures grown in mMRS by solvent extraction, gel filtration, RP-C(8) chromatography, and anion-exchange chromatography, followed by rechromatography by reversed-phase high-pressure liquid chromatography. Reutericyclin was characterized as a negatively charged, highly hydrophobic molecule with a molecular mass of 349 Da. Structural characterization (A. Holtzel, M. G. Ganzle, G. J. Nicholson, W. P. Hammes, and G. Jung, Angew. Chem. Int. Ed. 39:2766-2768, 2000) revealed that reutericyclin is a novel tetramic acid derivative. The inhibitory activity of culture supernatant of L. reuteri LTH2584 corresponded to that of purified as well as synthetic reutericyclin. PMID- 11010878 TI - Involvement of an extracellular protease in algicidal activity of the marine bacterium Pseudoalteromonas sp. strain A28. AB - The marine bacterium Pseudoalteromonas sp. strain A28 was able to kill the diatom Skeletonema costatum strain NIES-324. The culture supernatant of strain A28 showed potent algicidal activity when it was applied to a paper disk placed on a lawn of S. costatum NIES-324. The condensed supernatant, which was prepared by subjecting the A28 culture supernatant to ultrafiltration with a 10,000-M(w) cutoff membrane, showed algicidal activity, suggesting that strain A28 produced extracellular substances capable of killing S. costatum cells. The condensed supernatant was then found to have protease and DNase activities. Two Pseudoalteromonas mutants lacking algicidal activity, designated NH1 and NH2, were selected after N-methyl-N'-nitrosoguanidine mutagenesis. The culture supernatants of NH1 and NH2 showed less than 15% of the protease activity detected with the parental strain, A28. The protease was purified to homogeneity from A28 culture supernatants by using ion-exchange chromatography followed by preparative gel electrophoresis. Paper-disk assays revealed that the purified protease had potent algicidal activity. The purified protease had a molecular mass for 50 kDa, and the N-terminal amino acid sequence was determined to be Ala Thr-Pro-Asn-Asp-Pro. The optimum pH and temperature of the protease were found to be 8.8 and 30 degrees C, respectively, by using succinyl-Ala-Ala-Pro-Phe-p nitroanilide as a substrate. The protease activity was strongly inhibited by phenylmethylsulfonyl fluoride, diisopropyl fluorophosphate, antipain, chymostatin, and leupeptin. No significant inhibition was detected with EDTA, EGTA, phenanthroline or tetraethylenepentamine. These results suggest that Pseudoalteromonas sp. strain A28 produced an extracellular serine protease which was responsible for the algicidal activity of this marine bacterium. PMID- 11010879 TI - Molecular characterization of Yarrowia lipolytica and Candida zeylanoides isolated from poultry. AB - Yeast isolates from raw and processed poultry products were characterized using PCR amplification of the internally transcribed spacer (ITS) 5.8S ribosomal DNA region (ITS-PCR), restriction analysis of amplified products, randomly amplified polymorphic DNA (RAPD) analysis, and pulsed-field gel electrophoresis (PFGE). ITS PCR resulted in single fragments of 350 and 650 bp, respectively, from eight strains of Yarrowia lipolytica and seven strains of Candida zeylanoides. Digestion of amplicons with HinfI and HaeIII produced two fragments of 200 and 150 bp from Y. lipolytica and three fragments of 350, 150, and 100 bp from C. zeylanoides, respectively. Although these fragments showed species-specific patterns and confirmed species identification, characterization did not enable intraspecies typing. Contour-clamped heterogeneous electric field PFGE separated chromosomal DNA of Y. lipolytica into three to five bands, most larger than 2 Mbp, whereas six to eight bands in the range of 750 to 2,200 bp were obtained from C. zeylanoides. Karyotypes of both yeasts showed different polymorphic patterns among strains. RAPD analysis, using enterobacterial repetitive intergenic sequences as primers, discriminated between strains within the same species. Cluster analysis of patterns formed groups that correlated with the source of isolation. For ITS-PCR, extraction of DNA by boiling yeast cells was successfully used. PMID- 11010880 TI - Cold shock induction of thermal sensitivity in Listeria monocytogenes. AB - Cold shock at 0 to 15 degrees C for 1 to 3 h increased the thermal sensitivity of Listeria monocytogenes. In a model broth system, thermal death time at 60 degrees C was reduced by up to 45% after L. monocytogenes Scott A was cold shocked for 3 h. The duration of the cold shock affected thermal tolerance more than did the magnitude of the temperature downshift. The Z values were 8.8 degrees C for controls and 7.7 degrees C for cold-shocked cells. The D values of cold-shocked cells did not return to control levels after incubation for 3 h at 28 degrees C followed by heating at 60 degrees C. Nine L. monocytogenes strains that were cold shocked for 3 h exhibited D(60) values that were reduced by 13 to 37%. The D value reduction was greatest in cold-shocked stationary-phase cells compared to cells from cultures in either the lag or exponential phases of growth. In addition, cold-shocked cells were more likely to be inactivated by a given heat treatment than nonshocked cells, which were more likely to experience sublethal injury. The D values of chloramphenicol-treated control cells and chloramphenicol treated cold-shocked cells were no different from those of untreated cold-shocked cells, suggesting that cold shock suppresses synthesis of proteins responsible for heat protection. In related experiments, the D values of L. monocytogenes Scott A were decreased 25% on frankfurter skins and 15% in ultra-high temperature milk if the inoculated products were first cold shocked. Induction of increased thermal sensitivity in L. monocytogenes by thermal flux shows potential to become a practical and efficacious preventative control method. PMID- 11010881 TI - Cold shock and its effect on ribosomes and thermal tolerance in Listeria monocytogenes. AB - Differential scanning calorimetry (DSC) and fatty acid analysis were used to determine how cold shocking reduces the thermal stability of Listeria monocytogenes. Additionally, antibiotics that can elicit production of cold or heat shock proteins were used to determine the effect of translation blockage on ribosome thermal stability. Fatty acid profiles showed no significant variations as a result of cold shock, indicating that changes in membrane fatty acids were not responsible for the cold shock-induced reduction in thermal tolerance. Following a 3-h cold shock from 37 to 0 degrees C, the maximum denaturation temperature of the 50S ribosomal subunit and 70S ribosomal particle peak was reduced from 73.4 +/- 0.1 degrees C (mean +/- standard deviation) to 72.1 +/- 0.5 degrees C (P < or = 0.05), indicating that cold shock induced instability in the associated ribosome structure. The maximum denaturation temperature of the 30S ribosomal subunit peak did not show a significant shift in temperature (from 67.5 +/- 0.4 degrees C to 66.8 +/- 0.5 degrees C) as a result of cold shock, suggesting that either 50S subunit or 70S particle sensitivity was responsible for the intact ribosome fragility. Antibiotics that elicited changes in maximum denaturation temperature in ribosomal components also elicited reductions in thermotolerance. Together, these data suggest that ribosomal changes resulting from cold shock may be responsible for the decrease in D value observed when L. monocytogenes is cold shocked. PMID- 11010882 TI - PCR primers that amplify fungal rRNA genes from environmental samples. AB - Two PCR primer pairs were designed to amplify rRNA genes (rDNA) from all four major phyla of fungi: Ascomycota, Basidiomycota, Chytridomycota, and Zygomycota. PCRs performed with these primers showed that both pairs amplify DNA from organisms representing the major taxonomic groups of fungi but not from nonfungal sources. To test the ability of the primers to amplify fungal rDNA from environment samples, clone libraries from two avocado grove soils were constructed and analyzed. These soils possess different abilities to inhibit avocado root rot caused by Phythophthora cinnamomi. Analysis of the two rDNA clone libraries revealed differences in the two fungal communities. It also revealed a markedly different depiction of the soil fungal community than that generated by a culture-based analysis, confirming the value of rDNA-based approaches for identifying organisms that may not readily grow on agar media. Additional evidence of the usefulness of the primers was obtained by identifying fungi associated with avocado leaves. In both the soil and leaf analyses, no nonfungal rDNA sequences were identified, illustrating the selectivity of these PCR primers. This work demonstrates the ability of two newly developed PCR primer sets to amplify fungal rDNA from soil and plant tissue, thereby providing unique tools to examine this vast and mostly undescribed community of organisms. PMID- 11010883 TI - Bacterial functional redundancy along a soil reclamation gradient. AB - A strategy to measure bacterial functional redundancy was developed and tested with soils collected along a soil reclamation gradient by determining the richness and diversity of bacterial groups capable of in situ growth on selected carbon substrates. Soil cores were collected from four sites along a transect from the Jamari tin mine site in the Jamari National Forest, Rondonia, RO, Brazil: denuded mine spoil, soil from below the canopy of invading pioneer trees, revegetated soil under new growth on the forest edge, and the forest floor of an adjacent preserved forest. Bacterial population responses were analyzed by amending these soil samples with individual carbon substrates in the presence of bromodeoxyuridine (BrdU). BrdU-labeled DNA was then subjected to a 16S-23S rRNA intergenic analysis to depict the actively growing bacteria from each site. The number and diversity of bacterial groups responding to four carbon substrates (L serine, L-threonine, sodium citrate, and alpha-lactose hydrate) increased along the reclamation-vegetation gradient such that the preserved forest soil samples contained the highest functional redundancy for each substrate. These data suggest that bacterial functional redundancy increases in relation to the regrowth of plant communities and may therefore represent an important aspect of the restoration of soil biological functionality to reclaimed mine spoils. They also suggest that bacterial functional redundancy may be a useful indicator of soil quality and ecosystem functioning. PMID- 11010884 TI - Antisense downregulation of sigma(32) as a transient metabolic controller in Escherichia coli: effects on yield of active organophosphorus hydrolase. AB - Plasmids containing an antisense fragment of the sigma(32) gene were constructed and introduced into Escherichia coli cells. Downregulation of the sigma(32) mediated stress response was evaluated under heat shock and ethanol stress and during the production of organophosphorus hydrolase (OPH). Northern blot analyses revealed that sigma(32) sense mRNA was virtually undetected in antisense producing cultures from 5 to 20 min after antisense induction. However, lower molecular-weight bands were found, presumably due to partial degradation of sigma(32) mRNA. While a >10-fold increase in sigma(32) protein level was found under ethanol stress in the control cultures, antisense producing cultures resulted in a <3-fold increase, indicating downregulation of sigma(32). Correspondingly, antisense synthesis resulted in a decreased level of a sigma(32) regulated chaperone (GroEL) for the first 2 h after induction relative to control cultures without sigma(32) antisense mRNA. The total yield of OPH in the presence of sigma(32) antisense was, on average, 62% of the yield without antisense. However, during sigma(32) antisense production, a sixfold-higher specific OPH activity was observed compared to non-antisense-producing cultures. PMID- 11010885 TI - Bacterial origin and community composition in the barley phytosphere as a function of habitat and presowing conditions. AB - An understanding of the factors influencing colonization of the rhizosphere is essential for improved establishment of biocontrol agents. The aim of this study was to determine the origin and composition of bacterial communities in the developing barley (Hordeum vulgare) phytosphere, using denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes amplified from extracted DNA. Discrete community compositions were identified in the endorhizosphere, rhizoplane, and rhizosphere soil of plants grown in an agricultural soil for up to 36 days. Cluster analysis revealed that DGGE profiles of the rhizoplane more closely resembled those in the soil than the profiles found in the root tissue or on the seed, suggesting that rhizoplane bacteria primarily originated from the surrounding soil. No change in bacterial community composition was observed in relation to plant age. Pregermination of the seeds for up to 6 days improved the survival of seed-associated bacteria on roots grown in soil, but only in the upper, nongrowing part of the rhizoplane. The potential occurrence of skewed PCR amplification was examined, and only minor cases of PCR bias for mixtures of two different DNA samples were observed, even when one of the samples contained plant DNA. The results demonstrate the application of culture-independent, molecular techniques in assessment of rhizosphere bacterial populations and the importance of the indigenous soil population in colonization of the rhizosphere. PMID- 11010886 TI - PCR-based identification of MAT-1 and MAT-2 in the Gibberella fujikuroi species complex. AB - All sexually fertile strains in the Gibberella fujikuroi species complex are heterothallic, with individual mating types conferred by the broadly conserved ascomycete idiomorphs MAT-1 and MAT-2. We sequenced both alleles from all eight mating populations, developed a multiplex PCR technique to distinguish these idiomorphs, and tested it with representative strains from all eight biological species and 22 additional species or phylogenetic lineages from this species complex. In most cases, either an approximately 800-bp fragment from MAT-2 or an approximately 200-bp fragment from MAT-1 is amplified. The amplified fragments cosegregate with mating type, as defined by sexual cross-fertility, in a cross of Fusarium moniliforme (Fusarium verticillioides). Neither of the primer pairs amplify fragments from Fusarium species such as Fusarium graminearum, Fusarium pseudograminearum, and Fusarium culmorum, which have, or are expected to have, Gibberella sexual stages but are thought to be relatively distant from the species in the G. fujikuroi species complex. Our results suggest that MAT allele sequences are useful indicators of phylogenetic relatedness in these and other Fusarium species. PMID- 11010887 TI - Concentration and detection of caliciviruses in water samples by reverse transcription-PCR. AB - Human caliciviruses (HuCVs) cause waterborne outbreaks of gastroenteritis. Standard indicators of a safe water supply do not adequately predict contamination of water by viruses, including HuCVs. We developed a method to concentrate and detect HuCVs in water samples by using a cultivable primate calicivirus (Pan-1) as a model. Viable Pan-1 was seeded in different types of water and then filtered with a 1MDS filter, eluted with beef extract (BE), and reconcentrated by polyethylene glycol (PEG) precipitation. The viruses in the final samples were tested by plaque assay or by reverse transcription (RT)-PCR following extraction of the RNA with Trizol. Pan-1 was more sensitive to high-pH treatment than poliovirus was; a pH 9.0 BE solution was found to recover 35% more viable Pan-1 than a pH 9.5 BE solution recovered. Pan-1 was recovered from small volumes of deionized, finished, ground, and surface waters at efficiencies of 94, 73, 67, and 64%, respectively, when samples were assayed after elution without further concentration. When larger volumes of water (up to 40 liters) were tested after elution and concentration with PEG, 38, 19, and 14% of the seeded Pan-1 were recovered from finished, ground, and surface waters, respectively. The limit of detection of Pan-1 by RT-PCR was estimated to be 0.75 to 1.5 PFU in 40 liters of finished water. This method may be adapted for monitoring HuCVs in drinking water and other types of water for public health safety. PMID- 11010888 TI - Bacterial phosphating of mild (unalloyed) steel. AB - Mild (unalloyed) steel electrodes were incubated in phosphate-buffered cultures of aerobic, biofilm-forming Rhodococcus sp. strain C125 and Pseudomonas putida mt2. A resulting surface reaction leading to the formation of a corrosion inhibiting vivianite layer was accompanied by a characteristic electrochemical potential (E) curve. First, E increased slightly due to the interaction of phosphate with the iron oxides covering the steel surface. Subsequently, E decreased rapidly and after 1 day reached -510 mV, the potential of free iron, indicating the removal of the iron oxides. At this point, only scattered patches of bacteria covered the surface. A surface reaction, in which iron was released and vivianite precipitated, started. E remained at -510 mV for about 2 days, during which the vivianite layer grew steadily. Thereafter, E increased markedly to the initial value, and the release of iron stopped. Changes in E and formation of vivianite were results of bacterial activity, with oxygen consumption by the biofilm being the driving force. These findings indicate that biofilms may protect steel surfaces and might be used as an alternative method to combat corrosion. PMID- 11010889 TI - Identification of collagen-binding proteins in Lactobacillus spp. with surface enhanced laser desorption/ionization-time of flight ProteinChip technology. AB - Biosurfactants produced by Lactobacillus fermentum RC-14, L. rhamnosus GR-1 and 36, and L. casei Shirota were found to contain proteins that bind to both collagen types III and VI, as determined by surface-enhanced laser desorption/ionization (SELDI)-time of flight mass spectrometry. Both collagen types III and VI immobilized on SELDI preactivated ProteinChip arrays detected several different sizes (2 to 48 kDa) of collagen-binding proteins. Overall, the RC-14-produced biosurfactant contained the greatest number of collagen-binding proteins (RC-14 > GR-1 > 36 > Shirota), including the mature form of a previously cloned 29-kDa collagen-binding protein (referred to in its mature 26-kDa form). Although biosurfactants isolated from L. casei Shirota and L. rhamnosus 36 and GR 1 also contain several collagen-binding proteins, they do not contain the 26-kDa collagen-binding protein. Together, these results demonstrate the utility of the SELDI system as a means of rapidly characterizing clinically important but complex biosurfactant solutions. PMID- 11010890 TI - Field evaluation of a semiautomated method for rapid and simple analysis of recreational water microbiological quality. AB - An early warning system using a rapid enzymatic semiautomated method suitable for fecal coliform detection in recreational waters within 8 h was developed further and evaluated in this study. This rapid method was compared to the standard method followed in the United Kingdom. We used 1,011 samples originating from 206 different locations in Wales. When we assessed the presence or absence of fecal coliforms, targeting very low levels of contamination, we obtained 83.9% agreement between the rapid method and the lauryl sulfate broth-membrane filtration technique, whereas direct confirmation of the samples processed by the rapid method showed 89. 3% agreement. Environmental enzymatic background activity was found to be the main limiting factor for this method. Owing to a specific and integrated handling of the results by the software of the instrument, the percentage of false-positive results (a consequence of enzymatic background) was successfully limited to 2.9% by the direct confirmation evaluation. However, 7.8% false-negative results due to "late-growers" had to be accepted in order to produce results within a working day. At present, the method can be used in a more conservative way to assess the environmental threshold of 100 CFU of fecal coliforms per 100 ml in recreational waters. The implications of our findings with regard to the applicability of rapid enzymatic methods are discussed. PMID- 11010891 TI - Correlations between morphological, molecular biological, and physiological characteristics in clinical and nonclinical isolates of Acanthamoeba spp. AB - Eleven Acanthamoeba isolates, obtained from Acanthamoeba keratitis patients, from contact lens cases of non-Acanthamoeba keratitis patients, from asymptomatic individuals, from necrotic tissue, and from tap water and two reference strains were investigated by morphological, molecular biological, and physiological means in order to discriminate clinically relevant and nonrelevant isolates. All clinically relevant isolates showed Acanthamoeba sp. group II morphology. 18S ribosomal DNA sequencing revealed sequence type T4 to be the most prevalent group among the isolates and also the group recruiting most of the pathogenic strains. Interestingly, within T4 the strains of no clinical relevance clustered together. Moreover, physiological properties appeared to be highly consistent with initial pathogenicity and with sequence clustering. Altogether, the results of our study indicate a correlation between the phylogenetic relationship and pathogenicity. PMID- 11010892 TI - Vesicle-mediated transfer of virulence genes from Escherichia coli O157:H7 to other enteric bacteria. AB - Membrane vesicles are released from the surfaces of many gram-negative bacteria during growth. Vesicles consist of proteins, lipopolysaccharide, phospholipids, RNA, and DNA. Results of the present study demonstrate that membrane vesicles isolated from the food-borne pathogen Escherichia coli O157:H7 facilitate the transfer of genes, which are then expressed by recipient Salmonella enterica serovar Enteritidis or E. coli JM109. Electron micrographs of purified DNA from E. coli O157:H7 vesicles showed large rosette-like structures, linear DNA fragments, and small open-circle plasmids. PCR analysis of vesicle DNA demonstrated the presence of specific genes from host and recombinant plasmids (hly, L7095, mobA, and gfp), chromosomal DNA (uidA and eaeA), and phage DNA (stx1 and stx2). The results of PCR and the Vero cell assay demonstrate that genetic material, including virulence genes, is transferred to recipient bacteria and subsequently expressed. The cytotoxicity of the transformed enteric bacteria was sixfold higher than that of the parent isolate (E. coli JM109). Utilization of the nonhost plasmid (pGFP) permitted the evaluation of transformation efficiency (ca. 10(3) transformants microg of DNA(-1)) and demonstrated that vesicles can deliver antibiotic resistance. Transformed E. coli JM109 cells were resistant to ampicillin and fluoresced a brilliant green. The role vesicles play in genetic exchange between different species in the environment or host has yet to be defined. PMID- 11010893 TI - MET17 and hydrogen sulfide formation in Saccharomyces cerevisiae. AB - Commercial isolates of Saccharomyces cerevisiae differ in the production of hydrogen sulfide (H(2)S) during fermentation, which has been attributed to variation in the ability to incorporate reduced sulfur into organic compounds. We transformed two commercial strains (UCD522 and UCD713) with a plasmid overexpressing the MET17 gene, which encodes the bifunctional O-acetylserine/O acetylhomoserine sulfhydrylase (OAS/OAH SHLase), to test the hypothesis that the level of activity of this enzyme limits reduced sulfur incorporation, leading to H(2)S release. Overexpression of MET17 resulted in a 10- to 70-fold increase in OAS/OAH SHLase activity in UCD522 but had no impact on the level of H(2)S produced. In contrast, OAS/OAH SHLase activity was not as highly expressed in transformants of UCD713 (0.5- to 10-fold) but resulted in greatly reduced H(2)S formation. Overexpression of OAS/OAH SHLase activity was greater in UCD713 when grown under low-nitrogen conditions, but the impact on reduction of H(2)S was greater under high-nitrogen conditions. Thus, there was not a good correlation between the level of enzyme activity and H(2)S production. We measured cellular levels of cysteine to determine the impact of overexpression of OAS/OAH SHLase activity on sulfur incorporation. While Met17p activity was not correlated with increased cysteine production, conditions that led to elevated cytoplasmic levels of cysteine also reduced H(2)S formation. Our data do not support the simple hypothesis that variation in OAS/OAH SHLase activity is correlated with H(2)S production and release. PMID- 11010894 TI - Adaptation of the nisin-controlled expression system in Lactobacillus plantarum: a tool to study in vivo biological effects. AB - The potential of lactic acid bacteria as live vehicles for the production and delivery of therapeutic molecules is being actively investigated today. For future applications it is essential to be able to establish dose-response curves for the targeted biological effect and thus to control the production of a heterologous biopeptide by a live lactobacillus. We therefore implemented in Lactobacillus plantarum NCIMB8826 the powerful nisin-controlled expression (NICE) system based on the autoregulatory properties of the bacteriocin nisin, which is produced by Lactococcus lactis. The original two-plasmid NICE system turned out to be poorly suited to L. plantarum. In order to obtain a stable and reproducible nisin dose-dependent synthesis of a reporter protein (beta-glucuronidase) or a model antigen (the C subunit of the tetanus toxin, TTFC), the lactococcal nisRK regulatory genes were integrated into the chromosome of L. plantarum NCIMB8826. Moreover, recombinant L. plantarum producing increasing amounts of TTFC was used to establish a dose-response curve after subcutaneous administration to mice. The induced serum immunoglobulin G response was correlated with the dose of antigen delivered by the live lactobacilli. PMID- 11010895 TI - Possible interactions within a methanotrophic-heterotrophic groundwater community able to transform linear alkylbenzenesulfonates. AB - The relationships and interactions within a methanotrophic-heterotrophic groundwater community were studied in a closed system (shake culture) in the presence of methane as the primary carbon and energy source and with the addition of the pure linear alkylbenzenesulfonate (LAS) congener 2-[4-(sulfophenyl)]decan as a cometabolic substrate. When cultured under different conditions, this community was shown to be a stable association, consisting of one obligate type II methanotroph and four or five heterotrophs possessing different nutritional and physiological characteristics. The results of experiments examining growth kinetics and nutritional relationships suggested that a number of complex interactions existed in the community in which the methanotroph was the only member able to grow on methane and to cometabolically initiate LAS transformation. These growth and metabolic activities of the methanotroph ensured the supply of a carbon source and specific nutrients which sustained the growth of four or five heterotrophs. In addition to the obligatory nutritional relationships between the methanotroph and heterotrophs, other possible interactions resulted in the modification of basic growth parameters of individual populations and a concerted metabolic attack on the complex LAS molecule. Most of these relationships conferred beneficial effects on the interacting populations, making the community adaptable to various environmental conditions and more efficient in LAS transformation than any of the individual populations alone. PMID- 11010896 TI - Identification and analysis of the polyhydroxyalkanoate-specific beta ketothiolase and acetoacetyl coenzyme A reductase genes in the cyanobacterium Synechocystis sp. strain PCC6803. AB - Synechocystis sp. strain PCC6803 possesses a polyhydroxyalkanoate (PHA)-specific beta-ketothiolase encoded by phaA(Syn) and an acetoacetyl-coenzyme A (CoA) reductase encoded by phaB(Syn). A similarity search of the entire Synechocystis genome sequence identified a cluster of two putative open reading frames (ORFs) for these genes, slr1993 and slr1994. Sequence analysis showed that the ORFs encode proteins having 409 and 240 amino acids, respectively. The two ORFs are colinear and most probably coexpressed, as revealed by sequence analysis of the promoter regions. Heterologous transformation of Escherichia coli with the two genes and the PHA synthase of Synechocystis resulted in accumulation of PHAs that accounted for up to 12.3% of the cell dry weight under high-glucose growth conditions. Targeted disruption of the above gene cluster in Synechocystis eliminated the accumulation of PHAs. ORFs slr1993 and slr1994 thus encode the PHA specific beta-ketothiolase and acetoacetyl-CoA reductase of Synechocystis and, together with the recently characterized PHA synthase genes in this organism (S. Hein, H. Tran, and A. Steinbuchel, Arch. Microbiol. 170:162-170, 1998), form the first complete PHA biosynthesis pathway known in cyanobacteria. Sequence alignment of all known short-chain-length PHA-specific acetoacetyl-CoA reductases also suggests an extended signature sequence, VTGXXXGIG, for this group of proteins. Phylogenetic analysis further places the origin of phaA(Syn) and phaB(Syn) in the gamma subdivision of the division Proteobacteria. PMID- 11010897 TI - Molecular genetic manipulation of truncated Cry1C protein synthesis in Bacillus thuringiensis to improve stability and yield. AB - Cry1 protoxins of Bacillus thuringiensis are insecticidal 135-kDa proteins synthesized and assembled into parasporal crystals during sporulation. After ingestion, these crystals dissolve in the midgut and active toxins with molecular masses of about 65-kDa are released from the N-terminal half of the molecule by midgut proteases. Direct synthesis of the toxin-containing N-terminal half of Cry1 molecules using recombinant DNA techniques results in a low level of unstable truncated proteins that do not crystallize. In the present study, inclusions of truncated Cry1C (Cry1C-t) were obtained by combining genetic elements from other endotoxin genes and operons that enhance Cry protein synthesis and crystallization. Increased levels of Cry1C-t synthesis were achieved by using cyt1A promoters to drive expression of the 5' half of cry1C that included in the construct the 5' cry3A STAB-SD mRNA stabilizing sequence and the 3' stem-loop transcription terminator. RNA dot blot analysis showed that the STAB-SD and 3' transcriptional termination sequences were important for stabilization of truncated cry1C (cry1C-t) mRNA. A low level of cry1C-t mRNA was present when only the cyt1A promoters were used to express cry1C-t, but no accumulation of Cry1C-t was detected in Western blots. The orientation of the transcription terminator was important to enhancing Cry1C-t synthesis. Inclusion of the 20- and 29-kDa helper protein genes in cry1C-t constructs further enhanced synthesis. The Cry1C-t protein was toxic to Spodoptera exigua larvae, though the toxicity (50% lethal concentration [LC(50)] = 13.2 microg/ml) was lower than that of full-length Cry1C (LC(50) = 1.8 microg/ml). However, transformation of the HD1 isolate of B. thuringiensis subsp. kurstaki with the cry1C-t construct enhanced its toxicity to S. exigua as much as fourfold. PMID- 11010898 TI - Internal trehalose protects endocytosis from inhibition by ethanol in Saccharomyces cerevisiae. AB - Endocytosis in Saccharomyces cerevisiae is inhibited by concentrations of ethanol of 2 to 6% (vol/vol), which are lower than concentrations commonly present in its natural habitats. In spite of this inhibition, endocytosis takes place under enological conditions when high concentrations of ethanol are present. Therefore, it seems that yeast has developed some means to circumvent the inhibition. In this work we have investigated this possibility. We identified two stress conditions under which endocytosis was resistant to inhibition by ethanol: fermentation during nitrogen starvation and growth on nonfermentable substrates. Under these conditions, yeast accumulates stress protectors, primarily trehalose and Hsp104, a protein required for yeast to survive ethanol stress. We found the following. (i) The appearance of ethanol resistance was accompanied by trehalose accumulation. (ii) Mutant cells unable to synthesize trehalose also were unable to develop resistance. (iii) Mutant cells that accumulated trehalose during growth on sugars were resistant to ethanol even under this nonstressing condition. (iv) Mutant cells unable to synthesize Hsp104 were able to develop resistance. We conclude that trehalose is the major factor in the protection of endocytosis from ethanol. Our results suggest another important physiological role for trehalose in yeast. PMID- 11010899 TI - Microbial degradation of the multiply branched alkane 2,6,10,15,19, 23 hexamethyltetracosane (Squalane) by Mycobacterium fortuitum and Mycobacterium ratisbonense. AB - Among several bacterial species belonging to the general Gordonia, Mycobacterium, Micromonospora, Pseudomonas, and Rhodococcus, only two mycobacterial isolates, Mycobacterium fortuitum strain NF4 and the new isolate Mycobacterium ratisbonense strain SD4, which was isolated from a sewage treatment plant, were capable of utilizing the multiply branched hydrocarbon squalane (2,6,10,15,19, 23 hexamethyltetracosane) and its analogous unsaturated hydrocarbon squalene as the sole carbon source for growth. Detailed degradation studies and high-pressure liquid chromatography analysis showed a clear decrease of the concentrations of squalane and squalene during biomass increase. These results were supported by resting-cell experiments using strain SD4 and squalane or squalene as the substrate. The degradation of acyclic isoprenoids and alkanes as well as of acids derived from these compounds was also investigated. Inhibition of squalane and squalene degradation by acrylic acid indicated the possible involvement of beta oxidation in the degradation route. To our knowledge, this is the first report demonstrating the biodegradation of squalane by using defined axenic cultures. PMID- 11010901 TI - PCR assays that identify the grapevine dieback fungus Eutypa lata. AB - Eutypa lata is the causal fungal agent of Eutypa dieback, a serious grapevine necrotic disease. The erratic and delayed (1 to 2 months) appearance of characteristic conidia on culture media and the presence of numerous microorganisms in decaying wood make it difficult either to identify or to detect E. lata in grapevine wood samples. We designed six pairs of PCR primers for diagnosis of E. lata. Three primer pairs were derived from ribosomal DNA internal transcribed spacer sequences, and three pairs were derived from randomly amplified polymorphic DNA fragments. The six primer pairs could be used to amplify DNAs extracted from all of the E. lata isolates tested. They did not amplify DNAs from fungi and bacteria representing more than 50 different species of microorganisms associated with grapevine. We developed a simple protocol, leading to a rapid release of DNA, that enabled us to identify E. lata from pure or mixed cultures as well as from grapevine wood samples. Identification of E. lata in wood was achieved within a few hours, instead of the several weeks required for classical cultures on agar medium. We believe that the procedure described here can be adapted to detect other microorganisms involved in woody plant diseases. PMID- 11010900 TI - Geographical segregation of the neurotoxin-producing cyanobacterium Anabaena circinalis. AB - Blooms of the cyanobacterium Anabaena circinalis are a major worldwide problem due to their production of a range of toxins, in particular the neurotoxins anatoxin-a and paralytic shellfish poisons (PSPs). Although there is a worldwide distribution of A. circinalis, there is a geographical segregation of neurotoxin production. American and European isolates of A. circinalis produce only anatoxin a, while Australian isolates exclusively produce PSPs. The reason for this geographical segregation of neurotoxin production by A. circinalis is unknown. The phylogenetic structure of A. circinalis was determined by analyzing 16S rRNA gene sequences. A. circinalis was found to form a monophyletic group of international distribution. However, the PSP- and non-PSP-producing A. circinalis formed two distinct 16S rRNA gene clusters. A molecular probe was designed, allowing the identification of A. circinalis from cultured and uncultured environmental samples. In addition, probes targeting the predominantly PSP producing or non-PSP-producing clusters were designed for the characterization of A. circinalis isolates as potential PSP producers. PMID- 11010902 TI - Commensal interactions in a dual-species biofilm exposed to mixed organic compounds. AB - There is limited knowledge of interspecies interactions in biofilm communities. In this study, Pseudomonas sp. strain GJ1, a 2-chloroethanol (2-CE)-degrading organism, and Pseudomonas putida DMP1, a p-cresol-degrading organism, produced distinct biofilms in response to model mixed waste streams composed of 2-CE and various p-cresol concentrations. The two organisms maintained a commensal relationship, with DMP1 mitigating the inhibitory effects of p-cresol on GJ1. A triple-labeling technique compatible with confocal microscopy was used to investigate the influence of toxicant concentrations on biofilm morphology, species distribution, and exopolysaccharide production. Single-species biofilms of GJ1 shifted from loosely associated cell clusters connected by exopolysaccharide to densely packed structures as the p-cresol concentrations increased, and biofilm formation was severely inhibited at high p-cresol concentrations. In contrast, GJ1 was abundant when associated with DMP1 in a dual species biofilm at all p-cresol concentrations, although at high p-cresol concentrations it was present only in regions of the biofilm where it was surrounded by DMP1. Evidence in support of a commensal relationship between DMP1 and GJ1 was obtained by comparing GJ1-DMP1 biofilms with dual-species biofilms containing GJ1 and Escherichia coli ATCC 33456, an adhesive strain that does not mineralize p-cresol. Additionally, the data indicated that only tower-like cell structures in the GJ1-DMP1 biofilm produced exopolysaccharide, in contrast to the uniform distribution of EPS in the single-species GJ1 biofilm. PMID- 11010903 TI - Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments. AB - Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-) carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-10(5) CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well. PMID- 11010904 TI - Metabolic engineering of an aerobic sulfate reduction pathway and its application to precipitation of cadmium on the cell surface. AB - The conversion of sulfate to an excess of free sulfide requires stringent reductive conditions. Dissimilatory sulfate reduction is used in nature by sulfate-reducing bacteria for respiration and results in the conversion of sulfate to sulfide. However, this dissimilatory sulfate reduction pathway is inhibited by oxygen and is thus limited to anaerobic environments. As an alternative, we have metabolically engineered a novel aerobic sulfate reduction pathway for the secretion of sulfides. The assimilatory sulfate reduction pathway was redirected to overproduce cysteine, and excess cysteine was converted to sulfide by cysteine desulfhydrase. As a potential application for this pathway, a bacterium was engineered with this pathway and was used to aerobically precipitate cadmium as cadmium sulfide, which was deposited on the cell surface. To maximize sulfide production and cadmium precipitation, the production of cysteine desulfhydrase was modulated to achieve an optimal balance between the production and degradation of cysteine. PMID- 11010905 TI - Detection and identification of bacterial endosymbionts in arbuscular mycorrhizal fungi belonging to the family Gigasporaceae. AB - Intracellular bacteria have been found previously in one isolate of the arbuscular mycorrhizal (AM) fungus Gigaspora margarita BEG 34. In this study, we extended our investigation to 11 fungal isolates obtained from different geographic areas and belonging to six different species of the family Gigasporaceae. With the exception of Gigaspora rosea, isolates of all of the AM species harbored bacteria, and their DNA could be PCR amplified with universal bacterial primers. Primers specific for the endosymbiotic bacteria of BEG 34 could also amplify spore DNA from four species. These specific primers were successfully used as probes for in situ hybridization of endobacteria in G. margarita spores. Neighbor-joining analysis of the 16S ribosomal DNA sequences obtained from isolates of Scutellospora persica, Scutellospora castanea, and G. margarita revealed a single, strongly supported branch nested in the genus Burkholderia. PMID- 11010906 TI - A fruiting body tissue method for efficient Agrobacterium-mediated transformation of Agaricus bisporus. AB - We describe a modified Agrobacterium-mediated method for the efficient transformation of Agaricus bisporus. Salient features of this procedure include cocultivation of Agrobacterium and fruiting body gill tissue and use of a vector with a homologous promoter. This method offers new prospects for the genetic manipulation of this commercially important mushroom species. PMID- 11010907 TI - Bacterial activity in South Pole snow. AB - Large populations (200 to 5,000 cells ml(-1) in snowmelt) of bacteria were present in surface snow and firn from the south pole sampled in January 1999 and 2000. DNA isolated from this snow yielded ribosomal DNA sequences similar to those of several psychrophilic bacteria and a bacterium which aligns closely with members of the genus Deinococcus, an ionizing-radiation- and desiccation resistant genus. We also obtained evidence of low rates of bacterial DNA and protein synthesis which indicates that the organisms were metabolizing at ambient subzero temperatures (-12 to -17 degrees C). PMID- 11010908 TI - Use of combined microautoradiography and fluorescence in situ hybridization to determine carbon metabolism in mixed natural communities of uncultured bacteria from the genus Achromatium. AB - Combined microautoradiography and fluorescence in situ hybridization (FISH) was used to investigate carbon metabolism in uncultured bacteria from the genus Achromatium. All of the Achromatium species identified in a freshwater sediment from Rydal Water, Cumbria, United Kingdom, which were distinguishable only by FISH, assimilated both [(14)C]bicarbonate and [(14)C]acetate. This extends previous findings that Achromatium spp. present at another location could only utilize organic carbon sources. Achromatium spp., therefore, probably exhibit a range of physiologies, i.e., facultative chemolithoautotrophy, mixotrophy, and chemoorganoheterotrophy, similar to other large sulfur bacteria (e.g., Beggiatoa spp.). PMID- 11010909 TI - Development of 16S rRNA-based probes for the Coriobacterium group and the Atopobium cluster and their application for enumeration of Coriobacteriaceae in human feces from volunteers of different age groups. AB - Two 16S rRNA-targeted probes were developed: one for the Coriobacterium group and the other for the Atopobium cluster (which comprises most of the Coriobacteriaceae species, including the Coriobacterium group). The new probes were based on sequences of three new Coriobacteriaceae strains isolated from human feces and clinical material and sequences from databases. Application of the probes to fecal samples showed that formula-fed infants had higher numbers of Coriobacterium group cells in their feces than breast-fed infants. In addition, based on the presented results, it is hypothesized that with the increasing age of a person, the diversity of Atopobium cluster species present in the feces increases. PMID- 11010910 TI - Cometabolic degradation of dibenzofuran by biphenyl-cultivated Ralstonia sp. strain SBUG 290. AB - Cells of the gram-negative bacterium Ralstonia sp. strain SBUG 290 grown in the presence of biphenyl are able to cooxidize dibenzofuran which has been 1,2 hydroxylated. Meta cleavage of the 1, 2-dihydroxydibenzofuran between carbon atoms 1 and 9b produced 2-hydroxy-4-(3'-oxo-3'H-benzofuran-2'-yliden)but-2-enoic acid, which was degraded completely via salicylic acid. The presence of these intermediates indicates a degradation mechanism for dibenzofuran via lateral dioxygenation by Ralstonia sp. strain SBUG 290. PMID- 11010911 TI - Pacific Northwest marine sediments contain ammonia-oxidizing bacteria in the beta subdivision of the Proteobacteria. AB - The diversity of ammonia-oxidizing bacteria in aquatic sediments was studied by retrieving ammonia monooxygenase and methane monooxygenase gene sequences. Methanotrophs dominated freshwater sediments, while beta-proteobacterial ammonia oxidizers dominated marine sediments. These results suggest that gamma proteobacteria such as Nitrosococcus oceani are minor members of marine sediment ammonia-oxidizing communities. PMID- 11010912 TI - Sulfur regulation of the sulfate transporter genes sutA and sutB in Penicillium chrysogenum. AB - Penicillium chrysogenum uses sulfate as a source of sulfur for the biosynthesis of penicillin. Sulfate uptake and the mRNA levels of the sulfate transporter encoding sutB and sutA genes are all reduced by high sulfate concentrations and are elevated by sulfate starvation. In a high-penicillin-yielding strain, sutB is effectively transcribed even in the presence of excess sulfate. This deregulation may facilitate the efficient incorporation of sulfur into cysteine and penicillin. PMID- 11010913 TI - Improved template preparation for PCR-based assays for detection of food-borne bacterial pathogens. AB - Shigella flexneri, Salmonella enterica serotype Typhimurium, and Listeria monocytogenes were applied to FTA filters, and the filters were used directly as templates to demonstrate their sensitivity and applicability in PCR-based detection assays. With pure cultures, the sensitivities of detection by FTA filter-based PCR were 30 to 50 and 200 CFU for the gram-negative enterics and Listeria, respectively. Different numbers of S. flexneri cells were used in controlled contamination experiments with several different foods (produce, beef, and apple cider). Aliquots from concentrated food washes subsequently spotted onto FTA filters and assayed by PCR gave consistently positive results and detection limits similar to those observed with pure-culture dilutions. This universal method for PCR template preparation from bacterial cells is rapid and highly sensitive and reduces interference from food-associated inhibitors of PCR. In addition, its broad applicability eliminates the need for multiple methods for analysis of food matrices. PMID- 11010914 TI - Microscale diversity of the genus Nitrobacter in soil on the basis of analysis of genes encoding rRNA. AB - We looked at the diversity of [NO(2)](-) oxidizers at field scale by examining isolates at clump scale and in microsamples of soil (diameter, 50 microm). The genetic distances (as determined by amplified ribosomal DNA restriction analysis performed with Nitrobacter-specific primers) in a small clump of soil were as large as those between reference strains from large geographical areas. Diversity in individual microsamples was shown by serotyping. PMID- 11010915 TI - The presence of humic substances and DNA in RNA extracts affects hybridization results. AB - RNA extracts obtained from environmental samples are frequently contaminated with coextracted humic substances and DNA. It was demonstrated that the response in rRNA-targeted oligonucleotide probe hybridizations decreased as the concentrations of humic substances and DNA in RNA extracts increased. The decrease in hybridization signal in the presence of humic substances appeared to be due to saturation of the hybridization membrane with humic substances, resulting in a lower amount of target rRNA bound to the membrane. The decrease in hybridization response in the presence of low amounts of DNA may be the result of reduced rRNA target accessibility. The presence of high amounts of DNA in RNA extracts resulted in membrane saturation. Consistent with the observations for DNA contamination, the addition of poly(A) to RNA extracts, a common practice used to prepare RNA dilutions for membrane blotting, also reduced hybridization signals, likely because of reduced target accessibility and membrane saturation effects. PMID- 11010916 TI - Rapid and simple determination of the Escherichia coli phylogenetic group. AB - Phylogenetic analysis has shown that Escherichia coli is composed of four main phylogenetic groups (A, B1, B2, and D) and that virulent extra-intestinal strains mainly belong to groups B2 and D. Actually, phylogenetic groups can be determined by multilocus enzyme electrophoresis or ribotyping, both of which are complex, time-consuming techniques. We describe a simple and rapid phylogenetic grouping technique based on triplex PCR. The method, which uses a combination of two genes (chuA and yjaA) and an anonymous DNA fragment, was tested with 230 strains and showed excellent correlation with reference methods. PMID- 11010917 TI - Structure and species composition of mercury-reducing biofilms. AB - Mercury-reducing biofilms from packed-bed bioreactors treating nonsterile industrial effluents were shown to consist of a monolayer of bacteria by scanning electron microscopy. Droplets of several micrometers in diameter which accumulated outside of the bacterial cells were identified as elemental mercury by electron-dispersive X-ray analysis. The monospecies biofilms of Pseudomonas putida Spi3 initially present were invaded by additional strains, which were identified to the species level by thermogradient gel electrophoresis (TGGE) and 16S rDNA sequencing. TGGE community fingerprints of the biofilms showed that they were composed of the effluent bacteria and did not contain uncultivable microorganisms. Of the 13 effluent bacterial strains, 2 were not mercury resistant, while all the others had resistance levels similar to or higher than the inoculant strain. PMID- 11010918 TI - mRNA detection by reverse transcription-PCR for monitoring viability over time in an Enterococcus faecalis viable but nonculturable population maintained in a laboratory microcosm. AB - The viable but nonculturable (VBNC) state is a survival strategy adopted by bacteria when they are exposed to hostile environmental conditions. It has been shown that VBNC forms of bacteria are no longer capable of growing on conventional bacteriological media but conserve pathogenic factors and/or genes. It is thus necessary to develop methods capable of detecting nonculturable bacteria and of establishing their viability when the microbiological quality of environments is monitored. In this study we demonstrated that a gene was expressed during the VBNC state in a low-nutrient-concentration microcosm through detection of Enterococcus faecalis pbp5 mRNA by reverse transcription-PCR over a 3-month period. The presence of mRNA correlated with metabolic activity and resuscitation capability, indicating the viability of the VBNC cells. PMID- 11010919 TI - Incorporation of protease K into larval insect membrane vesicles does not result in disruption of integrity or function of the pore-forming Bacillus thuringiensis delta-endotoxin. AB - Bacillus thuringiensis delta-endotoxins insert into the brush border membranes of insect larval cells to form ion channels. A possible interaction of these toxins with a cytoplasmic component was examined by preloading vesicles from insect larval cells with protease K followed by incubation with toxin. There was no evidence for toxin antigens smaller than the intact toxin in extracts of solubilized vesicles, nor was there an effect of the inclusion of protease K on either of two functional properties, the formation of toxin aggregates or of ion pores. These toxins, physically and functionally, appear to be confined to the membrane. PMID- 11010920 TI - A PCR assay To discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. AB - Our purpose was to develop a rapid, inexpensive method of diagnosing the source of fecal pollution in water. In previous research, we identified Bacteroides Prevotella ribosomal DNA (rDNA) PCR markers based on analysis. These markers length heterogeneity PCR and terminal restriction fragment length polymorphism distinguish cow from human feces. Here, we recovered 16S rDNA clones from natural waters that were close phylogenetic relatives of the markers. From the sequence data, we designed specific PCR primers that discriminate human and ruminant sources of fecal contamination. PMID- 11010921 TI - Phenylacetyl-coenzyme A is the true inducer of the phenylacetic acid catabolism pathway in Pseudomonas putida U. AB - Aerobic degradation of phenylacetic acid in Pseudomonas putida U is carried out by a central catabolism pathway (phenylacetyl-coenzyme A [CoA] catabolon core). Induction of this route was analyzed by using different mutants specifically designed for this objective. Our results revealed that the true inducer molecule is phenylacetyl-CoA and not other structurally or catabolically related aromatic compounds. PMID- 11010922 TI - Antisense silencing of the creA gene in Aspergillus nidulans. AB - Antisense expression of a portion of the gene encoding the major carbon catabolite repressor CREA in Aspergillus nidulans resulted in a substantial increase in the levels of glucose-repressible enzymes, both endogenous and heterologous, in the presence of glucose. The derepression effect was approximately one-half of that achieved in a null creA mutant. Unlike results for that mutant, however, growth parameters and colony morphology in the antisense transformants were not affected. PMID- 11010923 TI - Cross-resistance of pink bollworm (Pectinophora gossypiella) to Bacillus thuringiensis toxins. AB - Two strains of pink bollworm (Pectinophora gossypiella) selected in the laboratory for resistance to Bacillus thuringiensis toxin Cry1Ac had substantial cross-resistance to Cry1Aa and Cry1Ab but not to Cry1Bb, Cry1Ca, Cry1Da, Cry1Ea, Cry1Ja, Cry2Aa, Cry9Ca, H04, or H205. The narrow spectrum of resistance and the cross-resistance to activated toxin Cry1Ab suggest that reduced binding of toxin to midgut target sites could be an important mechanism of resistance. PMID- 11010924 TI - A rhamnolipid biosurfactant reduces cadmium toxicity during naphthalene biodegradation. AB - A model cocontaminated system was developed to determine whether a metal complexing biosurfactant, rhamnolipid, could reduce metal toxicity to allow enhanced organic biodegradation by a Burkholderia sp. isolated from soil. Rhamnolipid eliminated cadmium toxicity when added at a 10-fold greater concentration than cadmium (890 microM), reduced toxicity when added at an equimolar concentration (89 microM), and had no effect at a 10-fold smaller concentration (8.9 microM). The mechanism by which rhamnolipid reduces metal toxicity may involve a combination of rhamnolipid complexation of cadmium and rhamnolipid interaction with the cell surface to alter cadmium uptake. PMID- 11010925 TI - A bioluminescent whole-cell reporter for detection of 2, 4-dichlorophenoxyacetic acid and 2,4-dichlorophenol in soil. AB - A bioreporter was made containing a tfdRP(DII)-luxCDABE fusion in a modified mini Tn5 construct. When it was introduced into the chromosome of Ralstonia eutropha JMP134, the resulting strain, JMP134-32, produced a sensitive bioluminescent response to 2, 4-dichlorophenoxyacetic acid (2,4-D) at concentrations of 2.0 microM to 5.0 mM. This response was linear (R(2) = 0.9825) in the range of 2.0 microM to 1.1 x 10(2) microM. Saturation occurred at higher concentrations, with maximal bioluminescence occurring in the presence of approximately 1.2 mM 2,4-D. A sensitive response was also recorded in the presence of 2,4-dichlorophenol at concentrations below 1.1 x 10(2) microM; however, only a limited bioluminescent response was recorded in the presence of 3-chlorobenzoic acid at concentrations below 1.0 mM. A significant bioluminescent response was also recorded when strain JMP134-32 was incubated with soils containing aged 2,4-D residues. PMID- 11010926 TI - Antibacterial activity evaluation of microcin J25 against diarrheagenic Escherichia coli. AB - The inhibitory activities of known microcins were evaluated against some diarrheagenic Escherichia coli strains. Some antibacterial properties of microcin J25, the most active one, were studied. A rapid two-step purification was performed. The MIC and the minimum bactericidal concentration of J25 against E. coli O157:H7 were 1 and 100 microg ml(-1), respectively. A 10(4)-CFU ml(-1) contamination by this strain was destroyed in milk and meat extract by 6.25 microg of J25 ml(-1) and in half-diluted egg yolk by 50 microg of J25 ml(-1). PMID- 11010927 TI - It's more than an apple a day: an appropriately processed, plant-centered dietary pattern may be good for your health. PMID- 11010928 TI - Colorful cancer prevention: alpha-carotene, lycopene, and lung cancer. PMID- 11010929 TI - Folate, DNA methylation, and gene expression: factors of nature and nurture. PMID- 11010930 TI - Differential oxidation of individual dietary fatty acids in humans. AB - BACKGROUND: Dietary fatty acids that are more prone to oxidation than to storage may be less likely to lead to obesity. OBJECTIVE: The aim of this study was to determine the effect of chain length, degree of unsaturation, and stereoisomeric effects of unsaturation on the oxidation of individual fatty acids in normal weight men. DESIGN: Fatty acid oxidation was examined in men consuming a weight maintenance diet containing 40% of energy as fat. After consuming the diet for 1 wk, subjects were fed fatty acids labeled with (13)C in the methyl or carboxyl position (10 mg/kg body wt). The fatty acids fed in random order were laurate, palmitate, stearate, oleate, elaidate (the trans isomer of oleate), linoleate, and linolenate blended in a hot liquid meal. Breath samples were collected for the next 9 h and the oxidation of each fatty acid was assessed by examining liberated (13)CO(2) in breath. RESULTS: Cumulative oxidation over the 9-h test ranged from a high of 41% of the dose for laurate to a low of 13% of the dose for stearate. Of the 18-carbon fatty acids, linolenate was the most highly oxidized and linoleate appeared to be somewhat conserved. (13)C recovery in breath from the methyl-labeled fatty acids was approximately 30% less than that from the carboxyl-labeled fatty acids. CONCLUSIONS: In summary, lauric acid is highly oxidized, whereas the polyunsaturated and monounsaturated fatty acids are fairly well oxidized. Oxidation of the long-chain, saturated fatty acids decreases with increasing carbon number. PMID- 11010931 TI - Prospective study of major dietary patterns and risk of coronary heart disease in men. AB - BACKGROUND: Previous studies on diet and coronary heart disease (CHD) focused primarily on individual nutrients or foods. OBJECTIVE: We examined whether overall dietary patterns derived from a food-frequency questionnaire (FFQ) predict risk of CHD in men. DESIGN: This was a prospective cohort study of 44875 men aged 40-75 y without diagnosed cardiovascular disease or cancer at baseline in 1986. RESULTS: During 8 y of follow-up, we documented 1089 cases of CHD (nonfatal myocardial infarction and fatal CHD). Using factor analysis, we identified 2 major dietary patterns using dietary data collected through a 131 item FFQ. The first factor, which we labeled the "prudent pattern," was characterized by higher intake of vegetables, fruit, legumes, whole grains, fish, and poultry, whereas the second factor, the "Western pattern," was characterized by higher intake of red meat, processed meat, refined grains, sweets and dessert, French fries, and high-fat dairy products. After adjustment for age and CHD risk factors, the relative risks from the lowest to highest quintiles of the prudent pattern score were 1.0, 0. 87, 0.79, 0.75, and 0.70 (95% CI: 0.56, 0.86; P: for trend = 0.0009). In contrast, the relative risks across increasing quintiles of the Western pattern score were 1.0, 1.21, 1.36, 1.40, and 1.64 (95% CI: 1.24, 2.17; P: for trend < 0.0001). These associations persisted in subgroup analyses according to cigarette smoking, body mass index, and parental history of myocardial infarction. CONCLUSIONS: These data suggest that major dietary patterns derived from the FFQ predict risk of CHD, independent of other lifestyle variables. PMID- 11010932 TI - Fruit and vegetable intake and risk of cardiovascular disease: the Women's Health Study. AB - BACKGROUND: Prospective data relating fruit and vegetable intake to cardiovascular disease (CVD) risk are sparse, particularly for women. OBJECTIVE: In a large, prospective cohort of women, we examined the hypothesis that higher fruit and vegetable intake reduces CVD risk. DESIGN: In 1993 we assessed fruit and vegetable intake among 39876 female health professionals with no previous history of CVD or cancer by use of a detailed food-frequency questionnaire. We subsequently followed these women for an average of 5 y for incidence of nonfatal myocardial infarction (MI), stroke, percutaneous transluminal coronary angioplasty, coronary artery bypass graft, or death due to CVD. RESULTS: During 195647 person-years of follow-up, we documented 418 incident cases of CVD including 126 MIs. After adjustment for age, randomized treatment status, and smoking, we observed a significant inverse association between fruit and vegetable intake and CVD risk. For increasing quintiles of total fruit and vegetable intake (median servings/d: 2. 6, 4.1, 5.5, 7.1, and 10.2), the corresponding relative risks (RRs) were 1.0 (reference), 0.78, 0.72, 0.68, and 0.68 (95% CI comparing the 2 extreme quintiles: 0.51, 0.92; P: for trend = 0.01). An inverse, though not statistically significant, trend remained after additional adjustment for other known CVD risk factors, with RRs of 1.0, 0.75, 0.83, 0.80, and 0.85 (95% CI for extreme quintiles: 0.61, 1.17). After excluding participants with a self-reported history of diabetes, hypertension, or high cholesterol at baseline, the multivariate-adjusted RR was 0.45 when extreme quintiles were compared (95% CI: 0.22, 0.91; P: for trend = 0.09). Higher fruit and vegetable intake was also associated with a lower risk of MI, with an adjusted RR of 0.62 for extreme quintiles (95% CI: 0.37, 1.04; P: for trend = 0.07). CONCLUSION: These data suggest that higher intake of fruit and vegetables may be protective against CVD and support current dietary guidelines to increase fruit and vegetable intake. PMID- 11010933 TI - Consumption of energy-dense, nutrient-poor foods by adult Americans: nutritional and health implications. The third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Current dietary guidance recommends limiting the intake of energy dense, nutrient-poor (EDNP) foods, but little is known about recent consumption patterns of these foods. OBJECTIVE: The contribution of EDNP foods to the American diet and the associated nutritional and health implications were examined. DESIGN: Data from the third National Health and Nutrition Examination Survey (n = 15611; age >/=20 y) were used. EDNP categories included visible fats, nutritive sweeteners and sweetened beverages, desserts, and snacks. The potential independent associations of EDNP food intake with intakes of energy, macronutrients, micronutrients, and serum vitamin, lipid, and carotenoid profiles were examined with linear and logistic regression procedures. RESULTS: EDNP foods supplied approximately 27% of energy intake; alcohol provided an additional 4%. The relative odds of consuming foods from all 5 food groups and of meeting the recommended dietary allowance or daily reference intake for protein and several micronutrients decreased with increasing EDNP food intake (P: < 0.0001). Energy intake and percentage of energy from fat were positively related to EDNP intake. Serum concentrations of vitamins A, E, C, and B-12; folate; several carotenoids; and HDL cholesterol were inversely related (P: +/- SEM: 7.5 +/- 1.2% with 28% carbohydrate, 9.2 +/- 1.5% with 53% carbohydrate, and 19.4 +/- 3.8% with 75% carbohydrate) and was nearly zero in a group of patients who had fasted for an average of 28 h (1.0 +/- 0.2%). In multiple regression analysis, DNL was correlated with carbohydrate intake, but not with body weight or plasma insulin concentrations. Endogenous glucose production, assessed with a dual-isotope technique, was not significantly different between the 3 groups of patients (13.7-15.3 micromol * kg(-1) * min(-1)), indicating impaired suppression by carbohydrate feeding. Gluconeogenesis was measured with [(13)C]bicarbonate, and increased as the carbohydrate intake increased (from 2.1 +/- 0.5 micromol * kg(-1) * min(-1) with 28% carbohydrate intake to 3.7 +/- 0.3 micromol * kg(-1) * min(-1) with 75% carbohydrate intake, P: < 0. 05). CONCLUSION: Carbohydrate feeding fails to suppress endogenous glucose production and gluconeogenesis, but stimulates DNL in critically ill patients. PMID- 11010936 TI - Energy metabolism after 2 y of energy restriction: the biosphere 2 experiment. AB - BACKGROUND: An adaptive decrease in energy expenditure (EE) in response to 6 mo of severely restricted energy intake was shown in a classic semistarvation study the Minnesota experiment. OBJECTIVE: Our objective was to examine whether such adaptation also occurs in response to less severe but sustained energy restriction. DESIGN: Body composition, 1-wk total EE (TEE), 24-h sedentary EE, and spontaneous physical activity were measured in 8 healthy subjects (4 men and 4 women) at the end of a 2-y confinement inside Biosphere 2. Unexpectedly, the food supply was markedly restricted during most of the confinement and all subjects experienced a marked, sustained weight loss (9.1 +/- 6.6 kg; P: < 0.001) from the low-energy (7000-11000 kJ/d), low-fat (9% of energy), but nutrient dense, diet they consumed. RESULTS: The TEE inside Biosphere 2, assessed 3 wk before exit, averaged 10700 +/- 560 kJ/d (n = 8). Within 1 wk after exit, the adjusted 24-h EE and spontaneous physical activity were lower in the biospherians (n = 5) than in 152 control subjects (6% and 45%, respectively; both P: < 0.01). Six months after exit and return to an ad libitum diet, body weight had increased to preentry levels; however, adjusted 24-h EE and spontaneous physical activity were still significantly lower than in control subjects. CONCLUSIONS: In lean humans, an adaptive decrease in EE appears to occur not only in states of life threatening undernutrition, but also in response to less severe energy restriction sustained over several years. PMID- 11010937 TI - Postprandial modulation of dietary and whole-body nitrogen utilization by carbohydrates in humans. AB - BACKGROUND: Sucrose exerts a sparing effect on whole-body protein metabolism, mainly during the absorptive phase. OBJECTIVE: We aimed to characterize the acute postprandial effect of addition of sucrose on deamination of dietary and endogenous nitrogen, with particular consideration being given to the effects of bioavailability. DESIGN: Twenty-one subjects equipped with ileal tubes ingested (15)N-labeled soy protein combined with [(13)C]glycine, with (n = 10) or without (n = 11) sucrose. Dietary and endogenous ileal flow of nitrogen were determined from the ileal effluents. The kinetics of dietary amino acid transfer to the blood were characterized by (13)CO(2) enrichment in breath and (15)N enrichment in plasma amino acids. Deamination of dietary and endogenous amino acid was determined from body urea, urinary nitrogen, and (15)N enrichment. RESULTS: (13)CO(2) recovery in breath and (15)N plasma amino acid enrichments were highly correlated (R:(2) >/= 0.95, P: < 0.001, for both meals) and markedly delayed by sucrose (half-(13)CO(2) recovery: 274 min compared with 167 min), whereas exogenous and endogenous ileal nitrogen kinetics and balances remained unchanged. Addition of sucrose halved the early (0-2 h) deamination peak of dietary nitrogen and reduced endogenous nitrogen oxidation over the first 4 h. Both were reduced by 18-24% over the 8-h period after the meal. CONCLUSIONS: Without changing the nitrogen absorptive balance, sucrose markedly affected the bioavailability profile, which is governed by gastric emptying. Endogenous and dietary nitrogen were not spared in the same way and over the same periods, showing that the metabolism of endogenous and dietary nitrogen may be affected differently by nutritional modulation, even if the effects are of a similar magnitude over the entire postprandial period. PMID- 11010938 TI - Iron deficiency anemia: higher prevalence in Mexican American than in non Hispanic white females in the third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Mexican American females have a higher prevalence of iron deficiency than do non-Hispanic white females. OBJECTIVE: The objective was to estimate the prevalence of iron deficiency anemia and examine potential reasons for this difference between Mexican American (n = 1194) and non-Hispanic white (n = 1183) females aged 12-39 y. DESIGN: We used data from the third National Health and Nutrition Examination Survey (1988-1994). Iron deficiency anemia was defined as abnormal results from >/=2 of 3 tests (erythrocyte protoporphyrin, transferrin saturation, and serum ferritin) and a low hemoglobin concentration. We used multiple logistic regression to adjust for factors that were more prevalent in Mexican American females and significantly associated with iron deficiency anemia. RESULTS: The prevalence of iron deficiency anemia was 6.2 +/- 0.8% (f1.gif" BORDER="0"> +/- SE) in Mexican American females and 2.3 +/- 0.4% in non Hispanic white females. Mean dietary iron intake, mean serum vitamin C concentrations, and the proportion of females using oral contraceptives were similar in the 2 groups. Age <20 y and education were not associated with iron deficiency anemia. After adjustment for poverty level, parity, and iron supplement use, the prevalence of iron deficiency anemia was 2.3 times higher in Mexican American than in non-Hispanic white females (95% CI: 1.4, 3.9). In those with a poverty income ratio (based on household income) >3.0, however, the prevalence of iron deficiency anemia was 2.6 +/- 0.9% in Mexican American and 1.9 +/- 0.6% in non-Hispanic white females (NS). CONCLUSION: Although much of the ethnic disparity in iron deficiency anemia remains unexplained, factors associated with household income may be involved. PMID- 11010939 TI - Use of vitamin-mineral supplements by female physicians in the United States. AB - BACKGROUND: Rates of vitamin-mineral supplement use by US female physicians are unknown but are of particular interest for several epidemiologic and clinical reasons. OBJECTIVE: The objective was to determine rates of and variations in vitamin-mineral supplement use among US female physicians. DESIGN: We used data from the Women Physicians' Health Study, a large (n = 4501) national, randomly sampled mail survey of female physicians aged 30-70 y. RESULTS: Half of the physicians took a multivitamin-mineral supplement; 35.5% of these did so regularly. However, /=6 mo after supplementation, except in subjects with low iron stores, whose serum ferritin returned to baseline within 3 mo. Fecal ferritin excretion increased 2.5-fold (P: < 0.05) during supplementation. CONCLUSIONS: Healthy individuals, even those with low iron stores, had reduced nonheme-iron absorption from food in response to iron supplementation. Despite this partial adaptation, iron stores were greater after iron supplementation than after placebo and this difference was sustained, except in individuals with the lowest iron stores. PMID- 11010942 TI - Intake of specific carotenoids and risk of lung cancer in 2 prospective US cohorts. AB - BACKGROUND: Carotenoids may reduce lung carcinogenesis because of their antioxidant properties; however, few studies have examined the relation between intakes of individual carotenoids and lung cancer risk. OBJECTIVE: The aim of this study was to examine the relation between lung cancer risk and intakes of alpha-carotene, beta-carotene, lutein, lycopene, and beta-cryptoxanthin in 2 large cohorts. DESIGN: During a 10-y follow-up period, 275 new cases of lung cancer were diagnosed in 46924 men; during a 12-y follow-up period, 519 new cases were diagnosed in 77283 women. Carotenoid intakes were derived from the reported consumption of fruit and vegetables on food-frequency questionnaires administered at baseline and during follow-up. The data were analyzed separately for each cohort and the results were pooled to compute overall relative risks (RRs). RESULTS: In the pooled analyses, alpha-carotene and lycopene intakes were significantly associated with a lower risk of lung cancer; the association with beta-carotene, lutein, and beta-cryptoxanthin intakes were inverse but not significant. Lung cancer risk was significantly lower in subjects who consumed a diet high in a variety of carotenoids (RR: 0.68; 95% CI: 0.49, 0.94 for highest compared with lowest total carotenoid score category). Inverse associations were strongest after a 4-8-y lag between dietary assessment and date of diagnosis. In subjects who never smoked, a 63% lower incidence of lung cancer was observed for the top compared with the bottom quintile of alpha-carotene intake (RR: 0.37; 95% CI: 0.18, 0.77). CONCLUSION: Data from 2 cohort studies suggest that several carotenoids may reduce the risk of lung cancer. PMID- 11010943 TI - Genomic DNA methylation decreases in response to moderate folate depletion in elderly women. AB - BACKGROUND: Methylation of genomic DNA is dependent on an adequate supply of folate coenzymes. Previous data support the hypothesis that abnormal DNA methylation plays an integral role in carcinogenesis. To date, no studies assessing the effect of inadequate folate status on DNA methylation in older women (aged >63 y) have been reported. OBJECTIVE: The effect of moderate folate depletion followed by folate repletion on leukocyte genomic DNA methylation was investigated in elderly women (aged 60-85 y) to evaluate whether DNA methylation could be used as a functional indicator of folate status. DESIGN: Healthy, postmenopausal women (n = 33) consumed a moderately folate-depleted diet (118 microg folate/d) for 7 wk, followed by 7 wk of folate repletion with 200 or 415 microg/d, each provided as 2 different dietary treatments for a total of 4 treatment groups (n = 30). Leukocyte DNA methylation was determined on the basis of the ability of DNA to incorporate [(3)H]methyl groups from labeled S: adenosylmethionine in an in vitro assay. RESULTS: Incorporation of [(3)H]methyl groups increased significantly (P: = 0.0025) in response to folate depletion, suggesting undermethylation of DNA. No significant changes were detected in [(3)H]methyl incorporation in any group over the 7-wk repletion period compared with postdepletion values. CONCLUSIONS: DNA methylation status may be used as a functional indicator of moderately depleted folate status. The slow response to the repletion diets observed suggests that normalization of DNA methylation after moderate folate depletion may be delayed in older women. PMID- 11010944 TI - Responsiveness of dark-adaptation threshold to vitamin A and beta-carotene supplementation in pregnant and lactating women in Nepal. AB - BACKGROUND: Impaired dark adaptation occurs commonly in vitamin A deficiency. OBJECTIVE: We sought to examine the responsiveness of dark-adaptation threshold to vitamin A and beta-carotene supplementation in Nepali women. DESIGN: The dark adapted pupillary response was tested in 298 pregnant women aged 15-45 y in a placebo-controlled trial of vitamin A and beta-carotene; 131 of these women were also tested at 3 mo postpartum. Results were compared with those for 100 nonpregnant US women of similar age. The amount of light required for pupillary constriction was recorded after bleaching and dark adaptation. RESULTS: Pregnant women receiving vitamin A had better dark-adaptation thresholds (-1.24 log cd/m(2)) than did those receiving placebo (-1.11 log cd/m(2); P: = 0. 03) or beta carotene (-1.13 log cd/m(2); P: = 0.05) (t tests with Bonferroni correction). Dark-adaptation threshold was associated with serum retinol concentration in pregnant women receiving placebo (P: = 0.001) and in those receiving beta carotene (P: = 0.003) but not in those receiving vitamin A. Among women receiving placebo, mean dark-adaptation thresholds were better during the first trimester ( 1.23 log cd/m(2)) than during the second and third trimesters (-1.03 log cd/m(2); P: = 0.02, t test). The mean threshold of nonpregnant US women (-1.35 log cd/m(2)) was better than that of all 3 Nepali groups (P: < 0.001, t test, for all 3 groups). CONCLUSIONS: During pregnancy, pupillary dark adaptation was strongly associated with serum retinol concentration and improved significantly in response to vitamin A supplementation. This noninvasive testing technique is a valid indicator of population vitamin A status in women of reproductive age. PMID- 11010945 TI - Infant growth patterns in the slums of Dhaka in relation to birth weight, intrauterine growth retardation, and prematurity. AB - BACKGROUND: Relations between size and maturity at birth and infant growth have been studied inadequately in Bangladesh, where the incidence of low birth weight is high and most infants are breast-fed. OBJECTIVE: This study was conducted to describe infant growth patterns and their relations to birth weight, intrauterine growth retardation, and prematurity. DESIGN: A total of 1654 infants born in selected low-socioeconomic areas of Dhaka, Bangladesh, were enrolled at birth. Weight and length were measured at birth and at 1, 3, 6, 9, and 12 mo of age. RESULTS: The infants' mean birth weight was 2516 g, with 46.4% weighing <2500 g; 70% were small for gestational age (SGA) and 17% were premature. Among the SGA infants, 63% had adequate ponderal indexes. The mean weight of the study infants closely tracked the -2 SD curve of the World Health Organization pooled breast fed sample. Weight differences by birth weight, SGA, or preterm categories were retained throughout infancy. Mean z scores based on the pooled breast-fed sample were -2.38, -1. 72, and -2.34 at birth, 3 mo, and 12 mo. Correlation analysis showed greater plasticity of growth in the first 3 mo of life than later in the first year. CONCLUSIONS: Infant growth rates were similar to those observed among breast-fed infants in developed countries. Most study infants experienced chronic intrauterine undernourishment. Catch-up growth was limited and weight at 12 mo was largely a function of weight at birth. Improvement of birth weight is likely to lead to significant gains in infant nutritional status in this population, although interventions in the first 3 mo are also likely to be beneficial. PMID- 11010946 TI - Tracking of body mass index from childhood to adolescence: a 6-y follow-up study in China. AB - BACKGROUND: Although extensive descriptive research shows that childhood obesity predisposes a person to adult obesity, little is understood about the dynamics of weight during childhood and the predictors of weight tracking. OBJECTIVE: Our objective was to examine tracking patterns of body mass index (BMI) as well as their predictors between childhood and adolescence. DESIGN: A cohort of 975 Chinese children aged 6-13 y was followed for 6 y (1991-1997). Tracking of BMI was defined as an individual maintaining a certain status (overweight or underweight) or relative position (relative BMI quartile) over time. Relative BMI related BMI to age- and sex-specific BMI cutoffs. RESULTS: After 6 y, approximately 40% of the subjects had maintained their relative positions, but 30% had moved into a lower or higher quartile. The BMIs of thin and fat children were more likely to track: 51% and 46% remained in the bottom and upper quartiles, respectively. Nearly one-third of the underweight children remained underweight in 1997. Overweight children were 2.8 times as likely as all other children to become overweight adolescents; underweight children were 3.6 times as likely to remain underweight as adolescents. Parental obesity and underweight, individuals' initial BMIs, dietary fat intake, and family income helped predict tracking and changes in BMI. CONCLUSION: In a society undergoing enormous changes in diet and activity, BMI tracking is still very important between childhood and adolescence in China. PMID- 11010947 TI - Energy expenditure of stunted and nonstunted boys and girls living in the shantytowns of Sao Paulo, Brazil. AB - BACKGROUND: Stunting increases the risk of obesity in developing countries, particularly in girls and women, but the underlying reason is not known. OBJECTIVE: Our objective was to test the hypothesis that stunted children have lower energy expenditure than do nonstunted children, a factor that has predicted an increased risk of obesity in other high-risk populations. DESIGN: A cross sectional study was conducted in shantytown children from Sao Paulo, Brazil. Twenty-eight stunted children aged 8-11 y were compared with 30 nonstunted children with similar weight-for-height. Free-living total energy expenditure (TEE) was measured over 7 d by using the doubly labeled water method. In addition, resting energy expenditure (REE) was measured by indirect calorimetry and body composition was measured by dual-energy X-ray absorptiometry. RESULTS: There were no significant associations between stunting and any measured energy expenditure parameter, including REE adjusted for weight (f1.gif" BORDER="0"> +/- SEM: 4575 +/- 95 compared with 4742 +/- 91 kJ/d, in stunted and nonstunted children, respectively) and TEE adjusted for weight (8424 +/- 239 compared with 8009 +/- 221 kJ/d, in stunted and nonstunted children, respectively). In multiple regression models that included fat-free mass and fat mass, girls had significantly lower TEE than did boys (P: < 0.05) but not significantly lower REE (P: = 0.17). CONCLUSIONS: There was no association between stunting and energy expenditure after differences between groups in body size and composition were accounted for. However, the girls had lower TEE than did boys, which may help to explain the particularly high risk of obesity in stunted adolescent girls and women in urban areas of developing countries. PMID- 11010948 TI - Prevalence and trends of overweight among preschool children in developing countries. AB - BACKGROUND: Obesity during childhood is a matter of growing concern. Several reports show increasing rates of obesity in developed countries, whereas the extent of the problem in developing countries remains unknown. OBJECTIVE: The aim of this study was to fill this gap by quantifying the prevalence and trends of overweight among preschool children in developing countries. DESIGN: One hundred sixty nationally representative cross-sectional surveys from 94 countries were analyzed in a standardized way to allow comparisons across countries and over time. Overweight was defined as a weight-for-height >2 SDs from the National Center for Health Statistics/World Health Organization international reference median. Prevalences of wasted children (< -2 SDs) are also presented to enable comparisons between both ends of the distribution. RESULTS: The global prevalence of overweight was 3.3%. Some countries and regions, however, had considerably higher rates, and overweight was shown to increase in 16 of 38 countries with trend data. Countries with the highest prevalences of overweight are located mainly in the Middle East, North Africa, and Latin America. Rates of wasting were generally higher than those of overweight; Africa and Asia had wasting rates 2.5 3.5 times higher than overweight rates. Countries with high wasting rates tended to have low overweight rates and vice versa. CONCLUSIONS: These estimates show that attention should be paid to monitoring levels and trends of overweight in children. This, however, should not be done at the expense of decreasing international commitments to alleviating undernutrition. The data presented confirm that undernutrition remains a major public health problem worldwide. PMID- 11010949 TI - How do fiber-supplemented formulas affect antroduodenal motility during enteral nutrition? A comparative study between mixed and insoluble fibers. AB - BACKGROUND: Fiber supplementation during enteral nutrition has been recommended, but the effect of soluble compared with insoluble fiber supplements on antroduodenal motility is unknown. OBJECTIVE: The objective of this study was to compare antroduodenal motor patterns in 8 healthy volunteers during and after gastric infusion of 3 different diets: a fiber-free diet, an insoluble-fiber diet, and a mixed-fiber diet (50% soluble fiber and 50% insoluble fiber). DESIGN: Manometric studies with the 3 different diets (2100 kJ) were performed in random order. Antroduodenal motility was monitored continuously for 6 h by using a pneumohydraulic system to calculate the number, amplitude, and duration of the pressure waves; the area under the curve (AUC); and the percentage of time occupied by motor activity before, during, and after each type of infusion. Variations in antral areas were measured by ultrasonography. RESULTS: The gastric motor response was significantly higher, whatever the diet, in the distal antral recording site than in the 2 more proximal sites. In the proximal but not the distal antrum, the number of waves, the AUC, and the percentage of time occupied by motor activity were higher (P: < 0.04) with the mixed-fiber than with the insoluble-fiber diet. No significant differences in variations of antral area were observed among the 3 diets. In the duodenum, motor variables were not significantly different among the 3 diets. CONCLUSIONS: A gastric infusion induced a greater motor response in the distal than in the proximal antrum. A mixed-fiber diet was associated with significantly greater proximal antral motility than was an insoluble-fiber diet. There was no significant difference among the 3 formulas in duodenal motor variables or in variations in antral area as measured by ultrasound. PMID- 11010950 TI - Garlic consumption and cancer prevention: meta-analyses of colorectal and stomach cancers. AB - BACKGROUND: Animal and in vitro studies have provided evidence of an anticarcinogenic effect of active ingredients in garlic. OBJECTIVE: The objective was to conduct meta-analyses of the epidemiologic literature on the association between garlic consumption and risk of stomach, colon, head and neck, lung, breast, and prostate cancers. DESIGN: Meta-analyses were conducted for all cancers mutually and separately for colorectal and stomach cancers in relation to consumption of exclusively raw garlic, cooked garlic, or both (RC garlic). Eighteen studies reported a relative risk estimate for RC garlic consumption and cancer risk. RESULTS: In the meta-analyses of colorectal and stomach cancer, the reference categories ranged from no consumption to consumption of 3.5 g/wk, whereas the highest categories ranged from any consumption to >28.8 g/wk. The average difference between the highest and lowest categories was 16 g/wk. The random-effects relative risk (RR) estimate of colorectal cancer and RC garlic consumption, excluding garlic supplements, was 0.69 (95% CI: 0.55, 0.89). For stomach cancer, the random-effects RR estimate was 0.53 (95% CI: 0.31, 0.92). The heterogeneity among studies for the latter outcome (P: = 0.0002) indicates the questionableness of the generalizability of this summary estimate. An indication of publication bias for all cancers combined is evident from a funnel plot of RC garlic consumption and cancer risk and from the results of the Begg and Mazumdar test (P: = 0.049). CONCLUSIONS: High intake of RC garlic may be associated with a protective effect against stomach and colorectal cancers. Heterogeneity of effect estimates, differences in dose estimation, publication bias, and possible alternative hypotheses (eg, confounding by total vegetable consumption) preclude sole reliance on summary effect estimates. PMID- 11010951 TI - Comparison of total body potassium with other techniques for measuring lean body mass in men and women with AIDS wasting. AB - BACKGROUND: Lean body mass is an important predictor of survival and functional status in patients with AIDS wasting. The bias between different techniques for assessing body composition in AIDS wasting is not known. DESIGN: We compared total body potassium (TBK) with fat-free mass (FFM) determined by dual-energy X ray absorptiometry (DXA), bioelectrical impedance analysis (BIA), and skinfold thickness measurement (SKF) in 132 patients (63 men, 69 women) with AIDS wasting (weight < 90% of ideal body weight, or weight loss > 10% of original, or both). None of the subjects exhibited clinical lipodystrophy. Comparisons were made by using different BIA equations. RESULTS: Lean body mass determined by DXA was highly correlated with TBK in men (r = 0.79, P: < 0.0001) and women (r = 0.84, P: < 0.0001). FFM(BIA) and FFM(DXA) were significantly different (P: < 0.01 in men and P: < 0.0001 in women). The difference between FFM(DXA) and FFM(BIA) was significantly greater with greater weight and body fat, particularly in HIV infected women (r = -0.39, P: = 0.001 for weight; r = -0.60, P: < 0.0001 for fat). The comparability of FFM and fat mass determined by DXA and BIA was dependent on the specific BIA equation used. Among men, no single BIA equation was more highly predictive of fat mass and FFM in comparison with DXA. CONCLUSIONS: The differences between DXA, BIA, and SKF in the determination of fat mass and FFM are significant in patients with AIDS wasting. BIA overestimates FFM compared with DXA in those with greater body fat. Standard BIA equations may not accurately estimate FFM and fat mass in men and women with AIDS wasting. PMID- 11010953 TI - No evidence for a link between consumption of chocolate and coronary heart disease. PMID- 11010956 TI - High-carbohydrate diets, physical activity, and plasma lipoprotein lipids. PMID- 11010957 TI - Breast-feeding, breast-milk feeding, and intelligence quotient. PMID- 11010958 TI - Physiologically meaningful biological models. PMID- 11010959 TI - Widespread distribution of binding sites for the novel Ca2+-mobilizing messenger, nicotinic acid adenine dinucleotide phosphate, in the brain. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca(2+) mobilizing agent in invertebrate eggs that has recently been shown to be active in certain mammalian and plant systems. Little, however, is known concerning the properties of putative NAADP receptors. Here, for the first time, we report binding sites for NAADP in brain. In contrast to sea urchin egg homogenates, [(32)P]NAADP bound reversibly to multiple sites in brain membranes. The rank order of potency of NAADP, 2',3'-cyclic NAADP and 3'-NAADP in displacing [(32)P]NAADP was, however, the same in the two systems and in agreement with their ability to mobilize Ca(2+) from homogenates. These data indicate that [(32)P]NAADP likely binds to receptors mediating Ca(2+) mobilization. Autoradiography revealed striking heterogeneity in the distribution of [(32)P]NAADP binding sites throughout the brain. Our data strongly support a role for NAADP-induced Ca(2+) signaling in the brain. PMID- 11010961 TI - Involvement of phospholipid hydroperoxide glutathione peroxidase in the modulation of prostaglandin D2 synthesis. AB - Antigenic cross-linking of the high affinity IgE receptors on mast cells induced the synthesis of prostaglandin D(2) (PGD(2)). The production of PGD(2) in L9 cells, which overexpressed non-mitochondrial phospholipid glutathione peroxidase (PHGPx), was only one-third that in the control line of cells (S1 cells). The reduction in the formation of PGD(2) in L9 cells was reversed upon inhibition of PHGPx activity by buthionine sulfoximine. Experiments with inhibitors demonstrated that prostaglandin H synthase-2 (PGHS-2) was the isozyme responsible for the production of PGD(2) upon cross-linking of IgE receptors. The conversion of radiolabeled arachidonic acid to prostaglandin H(2) (PGH(2)) was strongly inhibited in L9 cells, whereas the rate of conversion of PGH(2) to PGD(2) was the same in L9 cells and S1 cells, indicating that PGHS was inactivated in L9 cells. The PGHS activity in L9 cells was about half that in S1 cells. However, PGHS activity in L9 cells increased to the level in S1 cells upon the addition of the hydroperoxide 15-hydroperoxyeicosatetraenoic acid or of 3-chloroperoxybenzoic acid. These results suggest that non-mitochondrial PHGPx might be involved in the inactivation of PGHS-2 in nucleus and endoplasmic reticulum via reductions in levels of the hydroperoxides that are required for full activation of PGHS. Therefore, it appears that PHGPx might function as a modulator of the production of prostanoids, in addition to its role as an antioxidant enzyme. PMID- 11010960 TI - Identification of a novel neuromedin U receptor subtype expressed in the central nervous system. AB - Neuromedin U is a neuropeptide prominently expressed in the upper gastrointestinal tract and central nervous system. Recently, GPR66/FM-3 (NmU-R1) was identified as a specific receptor for neuromedin U. A BLAST search of the GenBank(TM) genomic database using the NmU-R1 cDNA sequence revealed a human genomic fragment encoding a G protein-coupled receptor that we designated NmU-R2 based on its homology to NmU-R1. The full-length NmU-R2 cDNA was subsequently cloned, stably expressed in 293 cells, and shown to mobilize intracellular calcium in response to neuromedin U. This response was dose-dependent (EC(50) = 5 nm) and specific in that other neuromedins did not induce a calcium flux in receptor-transfected cells. Expression analysis of human NmU-R2 demonstrated its mRNA to be most highly expressed in central nervous system tissues. Based on these data, we conclude that NmU-R2 is a novel neuromedin U receptor subtype that is likely to mediate central nervous system-specific neuromedin U effects. PMID- 11010962 TI - Interaction between the unphosphorylated receptor with high affinity for IgE and Lyn kinase. AB - Chinese hamster ovary fibroblasts previously transfected with the high affinity receptor for IgE (FcepsilonRI) were further transfected with the alpha subunit of the receptor for interleukin 2 (Tac) or with chimeric constructs in which the cytoplasmic domain of Tac was replaced with the C-terminal cytoplasmic domain of either the beta subunit or the gamma subunit of FcepsilonRI. Whereas native Tac failed to affect the aggregation-induced phosphorylation of FcepsilonRI, both chimeric constructs substantially inhibited this reaction. Alternatively, the FcepsilonRI-bearing fibroblasts were transfected with two chimeric constructs in which the cytoplasmic domain of Tac was replaced with a modified short form of Lyn kinase. The Lyn in both of the chimeric constructs had been mutated to remove the sites that are normally myristoylated and palmitoylated, respectively; one of the constructs had in addition been altered to be catalytically inactive. The catalytically active construct enhanced, and the inactive construct inhibited, aggregation-induced phosphorylation of the receptors. All of the chimeric constructs were largely distributed outside the detergent resistant microdomains, and whereas aggregation caused them to move to the domains in part, their aggregation was neither necessary nor enhanced their effects. These results and others indicate that the receptor and Lyn interact through protein-protein interactions that neither are dependent upon either the post-translational modification of the kinase with lipid moieties nor result exclusively from their co-localization in specialized membrane domains. PMID- 11010963 TI - Regulation of the ligand binding activity of the human very low density lipoprotein receptor by protein kinase C-dependent phosphorylation. AB - The very low density lipoprotein receptor (VLDL-R) binds and internalizes several ligands, including very low density lipoprotein (VLDL), urokinase-type plasminogen activator:plasminogen activator inhibitor type 1 complexes, lipoprotein lipase, and the 39-kDa receptor-associated protein that copurifies with the low density lipoprotein receptor-related protein/alpha(2)-macroglobulin receptor. Although several agonists regulate VLDL-R mRNA and/or protein expression, post-transcriptional regulation of receptor activity has not been described. Here, we report that the ligand binding activity of the VLDL-R in THP 1 monocytic cells, endothelial cells, smooth muscle cells, and VLDL-R-transfected HEK 293 cells is diminished after treatment with phorbol 12-myristate 13-acetate. This response was blocked by inhibitors of protein kinase C (PK-C), including a specific inhibitor of the PK-C beta II isoform, and was associated with phosphorylation of serine residues in the cytoplasmic domain of the receptor. Culture of endothelial cells in the presence of high glucose concentrations, which stimulate diacylglycerol synthesis and PK-C beta II activation, also induced a PK-C-dependent loss of VLDL-R ligand binding activity. Taken together, these studies demonstrate that the ligand binding activity of the VLDL-R is regulated by PK-C-dependent phosphorylation and that hyperglycemia may diminish VLDL-R activity. PMID- 11010964 TI - Molecular and functional characterization of organic cation/carnitine transporter family in mice. AB - Carnitine is essential for beta-oxidation of fatty acids, and a defect of cell membrane transport of carnitine leads to fatal systemic carnitine deficiency. We have already shown that a defect of the organic cation/carnitine transporter OCTN2 is a primary cause of systemic carnitine deficiency. In the present study, we further isolated and characterized new members of the OCTN family, OCTN1 and 3, in mice. All three members were expressed commonly in kidney, and OCTN1 and -2 were also expressed in various tissues, whereas OCTN3 was characterized by predominant expression in testis. When their cDNAs were transfected into HEK293 cells, the cells exhibited transport activity for carnitine and/or the organic cation tetraethylammonium (TEA). Carnitine transport by OCTN1 and OCTN2 was Na(+) dependent, whereas that by OCTN3 was Na(+)-independent. TEA was transported by OCTN1 and OCTN2 but not by OCTN3. The relative uptake activity ratios of carnitine to TEA were 1.78, 11.3, and 746 for OCTN1, -2, and -3, respectively, suggesting high specificity of OCTN3 for carnitine and significantly lower carnitine transport activity of OCTN1. Thus, OCTN3 is unique in its limited tissue distribution and Na(+)-independent carnitine transport, whereas OCTN1 efficiently transported TEA with minimal expression of carnitine transport activity and may have a different role from other members of the OCTN family. PMID- 11010965 TI - Selective degradation of oxidized calmodulin by the 20 S proteasome. AB - We have investigated the mechanisms that target oxidized calmodulin for degradation by the proteasome. After methionine oxidation within calmodulin, rates of degradation by the 20 S proteasome are substantially enhanced. Mass spectrometry was used to identify the time course of the proteolytic fragments released from the proteasome. Oxidized calmodulin is initially degraded into large proteolytic fragments that are released from the proteasome and subsequently degraded into small peptides that vary in size from 6 to 12 amino acids. To investigate the molecular determinants that result in the selective degradation of oxidized calmodulin, we used circular dichroism and fluorescence spectroscopy to assess oxidant-induced structural changes. There is a linear correlation between decreases in secondary structure and the rate of degradation. Calcium binding or the repair of oxidized calmodulin by methionine sulfoxide reductase induces comparable changes in alpha-helical content and rates of degradation. In contrast, alterations in the surface hydrophobicity of oxidized calmodulin do not alter the rate of degradation by the proteasome, indicating that changes in surface hydrophobicity do not necessarily lead to enhanced proteolytic susceptibility. These results suggest that decreases in secondary structure expose proteolytically sensitive sites in oxidized calmodulin that are cleaved by the proteasome in a nonprocessive manner. PMID- 11010966 TI - Molecular cloning of the human kallikrein 15 gene (KLK15). Up-regulation in prostate cancer. AB - Kallikreins are a subgroup of serine proteases with diverse physiological functions. Growing evidence suggests that many kallikreins are implicated in carcinogenesis. By using molecular cloning techniques, we identified a new human kallikrein gene, tentatively named KLK15 (for kallikrein 15 gene). This new gene maps to chromosome 19q13.4 and is located between the KLK1 and KLK3 genes. KLK15 is formed of five coding exons and four introns, and shows structural similarity to other kallikreins and kallikrein-like genes. KLK15 has three alternatively spliced forms and is primarily expressed in the thyroid gland and to a lower extent in the prostate, salivary, and adrenal glands and in the colon testis and kidney. Our preliminary results indicate that the expression of KLK15 is up regulated by steroid hormones in the LNCaP prostate cancer cell line. The KLK15 gene is also up-regulated, at the mRNA level, in prostate cancer in comparison to normal prostatic tissue. KLK15 up-regulation was found to be associated with more aggressive forms of prostate cancer. This newly discovered gene has the potential of being used as a diagnostic and/or prognostic marker for prostate cancer. PMID- 11010967 TI - Synergistic, p160 coactivator-dependent enhancement of estrogen receptor function by CARM1 and p300. AB - Members of the p160 coactivator family (steroid receptor coactivator-1 (SRC-1), glucocorticoid receptor interacting protein 1 (GRIP1), and activator of thyroid and retinoic acid receptors (ACTR)) mediate transcriptional activation by nuclear receptors. After being recruited to the promoter by nuclear receptors, the p160 coactivator transmits the activating signal via two C-terminal activation domains, AD1 and AD2. AD1 is a binding site for the related coactivators cAMP response element binding protein binding protein (CBP) and p300, whereas AD2 binds to another coactivator, coactivator-associated arginine methyltransferase 1 (CARM1), a protein-arginine methyltransferase. The current study explored the cooperative functional and mechanistic relationships among GRIP1, CARM1, and p300 in transient transfection assays, where they enhanced the ability of the estrogen receptor (ER) to activate transcription of a reporter gene. The coactivator functions of p300 and CARM1 depended on the co-expression of GRIP1. Simultaneous co-expression of all three coactivators caused a synergistic enhancement of ER function. Deletion of the AD1 domain of GRIP1 abolished the ability of p300 to potentiate ER activity but had no effect on CARM1-mediated stimulation. In contrast, when the AD2 domain of GRIP1 was deleted, p300 still stimulated ER function through the mutant GRIP1, but CARM1 failed to do so. Thus, both binding of p300 to AD1 and binding of CARM1 to AD2 are required for their respective coactivator functions and for their synergy. Furthermore, CARM1 and p300 function independently through different activating domains of GRIP1, and their synergy suggests that they enhance transcription by different, complementary mechanisms. PMID- 11010968 TI - Identification and characterization of a novel follistatin-like protein as a binding protein for the TGF-beta family. AB - Follistatin is an activin-binding protein that prevents activin from binding to its receptors and neutralizes its activity. Follistatin also binds bone morphogenetic proteins (BMPs). In this study, we report the identification of a novel follistatin-like protein from mouse. The mouse cDNA encodes a 256-residue precursor and most likely a mouse homologue of human FLRG, which was found at the breakpoint of the chromosomal rearrangement in a B-cell line. Whereas follistatin has three follistatin domains, which are presumed to be growth factor binding motifs, FLRG possesses only two follistatin domains. Northern blotting revealed that mRNAs for FLRG were abundantly expressed in heart, lung, kidney, and testis in mouse. The recombinant mouse FLRG proteins were found to have binding activity for both activin and bone morphogenetic protein-2. Like follistatin, FLRG has higher affinity for activin than for BMP-2. The FLRG protein inhibited activin induced and BMP-2-induced transcriptional responses in a dose-dependent manner. Glutathione S-transferase fusion proteins encoding various regions of FLRG were produced and studied. Ligand blotting using (125)I-activin revealed that the COOH terminal region containing the second follistatin domain was able to bind activin. Our finding implies that cellular signaling by activin and BMPs is tightly regulated by multiple members of the follistatin family. PMID- 11010969 TI - The isolation, characterization, and molecular cloning of a 75-kDa gelatinase B like enzyme, a member of the matrix metalloproteinase (MMP) family. An avian enzyme that is MMP-9-like in its cell expression pattern but diverges from mammalian gelatinase B in sequence and biochemical properties. AB - We have isolated a novel 75-kDa gelatinase from a chicken macrophage cell line, HD11. Biochemical and immunological characterization of the purified enzyme demonstrated that it is distinct from the chicken 72-kDa gelatinase A (MMP-2). The enzyme is capable of specific gelatin binding and rapid gelatin cleavage. Incubation with an organomercurial compound (p-aminophenylmercuric acetate) induces proteolytic processing and activation of this enzyme, and the resultant gelatinolytic activity is sensitive to both zinc chelators and tissue inhibitors of metalloproteinases. A full-length cDNA for the enzyme has been cloned, and sequence analysis demonstrated that the enzyme possesses the characteristic multidomain structure of an MMP gelatinase including a cysteine switch prodomain, three fibronectin type II repeats, a catalytic zinc binding region, and a hemopexin-like domain. The 75-kDa gelatinase is produced by phorbol ester-treated chicken bone marrow cells, monocytes, and polymorphonuclear leukocytes, cell types that charac- teristically produce the 92-kDa mammalian gelatinase B (MMP 9). The absence of a 90-110-kDa gelatinase in these cell types indicates that the 75-kDa gelatinase is likely the avian counterpart of gelatinase B. However, the protein is only 59% identical to human gelatinase B, whereas all previously cloned chicken MMP homologues are 75-90% identical to their human counterparts. In addition, the new 75-kDa chicken gelatinase lacks the type V collagen domain that is found in all mammalian gelatinase Bs. Furthermore, the secreted enzyme appears structurally distinct from known gelatinase Bs and the activated enzyme can cleave fibronectin, which is not a substrate for mammalian gelatinase B. Thus the results of this study indicate that a second MMP gelatinase exists in chickens, and although it is MMP-9/gelatinase B-like in its overall domain structure and expression pattern, it appears to be biochemically divergent from mammalian gelatinase B. PMID- 11010970 TI - Regulation of a Ca2+-sensitive adenylyl cyclase in an excitable cell. Role of voltage-gated versus capacitative Ca2+ entry. AB - In nonexcitable cells, we had previously established that Ca(2+)-sensitive adenylyl cyclases, whether expressed endogenously or heterologously, were regulated exclusively by capacitative Ca(2+) entry (Fagan, K. A., Mahey, R. and Cooper, D. M. F. (1996) J. Biol. Chem. 271, 12438-12444; Fagan, K. A., Mons, N., and Cooper, D. M. F. (1998) J. Biol. Chem. 273, 9297-9305). Relatively little is known about how these enzymes are regulated by Ca(2+) in excitable cells, where they predominate. Furthermore, no effort has been made to determine whether the prominent voltage-gated Ca(2+) entry, which typifies excitable cells, overwhelms the effect of any capacitative Ca(2+) entry that may occur. In the present study, we placed the Ca(2+)-stimulable, adenylyl cyclase type VIII in an adenovirus vector to optimize its expression in the pituitary-derived GH(4)C(1) cell line. In these cells, a modest degree of capacitative Ca(2+) entry could be discerned in the face of a dramatic voltage-gated Ca(2+) entry. Nevertheless, both modes of Ca(2+) entry were equally efficacious at stimulating adenylyl cyclase. A striking release of Ca(2+) from intracellular stores, triggered either by ionophore or thyrotrophin-releasing hormone, was incapable of stimulating the adenylyl cyclase. It thus appears as though the intimate colocalization of adenylyl cyclase with capacitative Ca(2+) entry channels is an intrinsic property of these molecules, regardless of whether they are expressed in excitable or nonexcitable cells. PMID- 11010971 TI - Alternative splicing in intracellular loop connecting domains II and III of the alpha 1 subunit of Cav1.2 Ca2+ channels predicts two-domain polypeptides with unique C-terminal tails. AB - Novel splice variants of the alpha(1) subunit of the Ca(v)1.2 voltage-gated Ca(2+) channel were identified that predicted two truncated forms of the alpha(1) subunit comprising domains I and II generated by alternative splicing in the intracellular loop region linking domains II and III. In rabbit heart splice variant 1 (RH-1), exon 19 was deleted, which resulted in a reading frameshift of exon 20 with a premature termination codon and a novel 19-amino acid carboxyl terminal tail. In the RH-2 variant, exons 17 and 18 were deleted, leading to a reading frameshift of exons 19 and 20 with a premature stop codon and a novel 62 amino acid carboxyl-terminal tail. RNase protection assays with RH-1 and RH-2 cRNA probes confirmed the expression in cardiac and neuronal tissue but not skeletal muscle. The deduced amino acid sequence from full-length cDNAs encoding the two variants predicted polypeptides of 99.0 and 99.2 kDa, which constituted domains I and II of the alpha(1) subunit of the Ca(v)1.2 channel. Antipeptide antibodies directed to sequences in the second intracellular loop between domains II and III identified the 240-kDa Ca(v)1.2 subunit in sarcolemmal and heavy sarcoplasmic reticulum (HSR) membranes and a 99-kDa polypeptide in the HSR. An antipeptide antibody raised against unique sequences in the RH-2 variant also identified a 99-kDa polypeptide in the HSR. These data reveal the expression of additional Ca(2+) channel structural units generated by alternative splicing of the Ca(v)1.2 gene. PMID- 11010973 TI - CHOP/GADD153 gene expression response to cellular stresses inhibited by prior exposure to ultraviolet light wavelength band C (UVC). Inhibitory sequence mediating the UVC response localized to exon 1. AB - CHOP/GADD153 is both an activating and repressing transcription factor that is markedly induced in response to a variety of cellular stresses. The CHOP/GADD153 gene was originally cloned because of its inducibility by ultraviolet light wavelength band C (UVC) and has since been found to be activated in response to many different cellular stresses. Some of the recent studies have questioned the UVC responsiveness of the CHOP gene. Contradiction in our own data led us to reexamine the UVC effects on CHOP expression. UVC is capable of strongly activating the mouse CHOP promoter in stably transfected NIH 3T3 cells but has only a modest and transient effect on the level of the CHOP messenger RNA. In addition to its positive effect on CHOP promoter activity, we show that UVC negatively affects CHOP mRNA and protein expression. Pretreatment of NIH 3T3 cells with UVC markedly attenuates the subsequent induction of CHOP mRNA by the cellular stress activators methylmethane sulfate, tunicamycin, glucose deprivation, and methionine deprivation for as long as at least 16 h. This inhibitory effect of UVC on CHOP expression in response to stress is independent of the presence or absence of p53 and does not involve mRNA degradation as opposed to the UVC effect that inhibits p21 expression seen only in the absence of p53. The target of the inhibitory effect of UVC on CHOP expression is located in the first exon of the gene, a 5'-untranslated region that is unusually conserved between different species. These findings suggest that an unknown function encoded by the 5'-untranslated region somehow modifies the response of CHOP gene transcription to UVC. PMID- 11010972 TI - BCR/ABL regulates expression of the cyclin-dependent kinase inhibitor p27Kip1 through the phosphatidylinositol 3-Kinase/AKT pathway. AB - Deregulation of cell cycle checkpoints is an almost universal abnormality in human cancers and is most often due to loss-of-function mutations of tumor suppressor genes such as Rb, p53, or p16(INK4a). In this study, we demonstrate that BCR/ABL inhibits the expression of a key cell cycle inhibitor, p27(Kip1), by signaling through a pathway involving phosphatidylinositol 3-kinase (PI3K). p27(Kip1) is a widely expressed inhibitor of cdk2, an essential cell cycle kinase regulating entry into S phase. We demonstrate that the decrease of p27(Kip1) is directly due to BCR/ABL in hematopoietic cells by two different approaches. First, induction of BCR/ABL by a tetracycline-regulated promoter is associated with a reversible down-regulation of p27(Kip1). Second, inhibition of BCR/ABL kinase activity with the Abl tyrosine kinase inhibitor STI571 rapidly increases p27(Kip1) levels. The PI3K inhibitor LY-294002 blocks the ability of BCR/ABL to induce p27(Kip1) down-regulation and inhibits BCR/ABL-induced entry into S phase. The serine/threonine kinase AKT/protein kinase B is a known downstream target of PI3K. Transient expression of an activated mutant of AKT was found to decrease expression of p27(Kip1), even when PI3K was inhibited by LY-294002. The mechanism of p27(Kip1) regulation is primarily related to protein stability, since inhibition of proteasome activity increased p27(Kip1) levels in BCR/ABL transformed cells, whereas very little change in p27 transcription was found. Overall, these data are consistent with a model in which BCR/ABL suppresses p27(Kip1) protein levels through PI3K/AKT, leading to accelerated entry into S phase. This activity is likely to explain in part previous studies showing that activation of PI3K was required for optimum transformation of hematopoietic cells by BCR/ABL in vitro and in vivo. PMID- 11010974 TI - Multiprotein complex formation at the beta myosin heavy chain distal muscle CAT element correlates with slow muscle expression but not mechanical overload responsiveness. AB - To examine the role of the beta-myosin heavy chain (betaMyHC) distal muscle CAT (MCAT) element in muscle fiber type-specific expression and mechanical overload (MOV) responsiveness, we conducted transgenic and in vitro experiments. In adult transgenic mice, mutation of the distal MCAT element led to significant reductions in chloramphenicol acetyltransferase (CAT) specific activity measured in control soleus and plantaris muscles when compared with wild type transgene beta293WT but did not abolish MOV-induced CAT specific activity. Electrophoretic mobility shift assay revealed the formation of a specific low migrating nuclear protein complex (LMC) at the betaMyHC MCAT element that was highly enriched only when using either MOV plantaris or control soleus nuclear extract. Scanning mutagenesis of the betaMyHC distal MCAT element revealed that only the nucleotides comprising the core MCAT element were essential for LMC formation. The proteins within the LMC when using either MOV plantaris or control soleus nuclear extracts were antigenically related to nominal transcription enhancer factor 1 (NTEF-1), poly(ADP-ribose) polymerase (PARP), and Max. Only in vitro translated TEF-1 protein bound to the distal MCAT element, suggesting that this multiprotein complex is tethered to the DNA via TEF-1. Protein-protein interaction assays revealed interactions between nominal TEF-1, PARP, and Max. Our studies show that for transgene beta293 the distal MCAT element is not required for MOV responsiveness but suggest that a multiprotein complex likely comprised of nominal TEF-1, PARP, and Max forms at this element to contribute to basal slow fiber expression. PMID- 11010975 TI - Germ-free and colonized mice generate the same products from enteric prodefensins. AB - The use of germ-free mice offers the possibility to study antibacterial components in a gut uncolonized by bacteria. We have developed a method to extract and high pressure liquid chromatography-fractionate the antibacterial factors present in the small intestine of a single mouse. By mass spectrometry and sequence analyses of fractions exhibiting antimicrobial activity, we identified and characterized the defensin region in germ-free mice as well as in colonized mice. Defensins made up around 15% of the total antibacterial activity both in germ-free and colonized mice. The intestine of germ-free mice exhibited the same set of mature enteric defensins (defensins 1, 2, 3, 4, and 6) as mice colonized by a normal microflora. Mature defensins are generated through processing of larger precursors by enzymatic removal of a signal peptide and a propiece. We found that all prodefensins were cleaved at a Ser/Ala-Leu bond, giving 34-residue propiece peptides and only trace amounts of the predicted 39 residue peptide. This first step must be followed by the removal of a residual peptide to render the mature defensins, indicating that the processing is more complex than previously anticipated. The same propieces were found in both germ free and colonized mice, suggesting that the same processing operates independent of bacterial presence in the intestine. PMID- 11010976 TI - Differential activation of p38 mitogen-activated protein kinase isoforms depending on signal strength. AB - We have investigated the ability of the mitogen-activated protein kinase (MAPK) kinase MKK6 to activate different members of the p38 subfamily of MAPKs and found that some MKK6 mutants can efficiently activate p38alpha but not p38gamma. In contrast, a constitutively active MKK6 mutant activated both p38 MAPK isoforms to similar extents. The same results were obtained upon co-expression in Xenopus oocytes and in vitro using either MKK6 immunoprecipitates from transfected cells or bacterially produced recombinant proteins. We also found that the preferential activation of p38alpha by MKK6 correlated with more efficient binding of MKK6 to p38alpha than to p38gamma. Furthermore, increasing concentrations of constitutively active MKK6 differentially activated either p38alpha alone (low MKK6 activity) or both p38alpha and p38gamma (high MKK6 activity), both in vitro and in injected oocytes. The determinants for selectivity are located at the carboxyl-terminal lobe of p38 MAPKs but do not correspond to the activation loop or common docking sequences. We also showed that different stimuli can induce different levels of endogenous MKK6 activity that correlate with differential activation of p38 MAPKs. Our results suggest that the level of MKK6 activity triggered by a given stimulus may determine the pattern of downstream p38 MAPK activation in the particular response. PMID- 11010977 TI - NMR analysis of type III antifreeze protein intramolecular dimer. Structural basis for enhanced activity. AB - The structure of a new antifreeze protein (AFP) variant, RD3, from antarctic eel pout (Rhigophila dearborni) with enhanced activity has been determined for the first time by nuclear magnetic resonance spectroscopy. RD3 comprises a unique translational topology of two homologous type III AFP globular domains, each containing one flat, ice binding plane. The ice binding plane of the N domain is located approximately 3.5 A "behind" that of the C domain. The two ice binding planes are located laterally with an angle of 32 +/- 12 degrees between the planes. These results suggest that the C domain plane of RD3 binds first to the ice [1010] prism plane in the <0001> direction, which induces successive ice binding of the N domain in the <0101> direction. This manner of ice binding caused by the unique structural topology of RD3 is thought to be crucial for the significant enhancement of antifreeze activity, especially at low AFP concentrations. PMID- 11010978 TI - Modulation of amyloid precursor protein metabolism by X11alpha /Mint-1. A deletion analysis of protein-protein interaction domains. AB - Modulation of amyloid precursor protein (APP) metabolism plays a pivotal role in the pathogenesis of Alzheimer's disease. The phosphotyrosine-binding/protein interaction (PTB/PI) domain of X11alpha, a neuronal cytosolic adaptor protein, binds to the YENPTY sequence in the cytoplasmic carboxyl terminus of APP. This interaction prolongs the half-life of APP and inhibits Abeta40 and Abeta42 secretion. X11alpha/Mint-1 has multiple protein-protein interaction domains, a Munc-18 interaction domain (MID), a Cask/Lin-2 interaction domain (CID), a PTB/PI domain, and two PDZ domains. These X11alpha protein interaction domains may modulate its effect on APP processing. To test this hypothesis, we performed a deletion analysis of X11alpha effects on metabolism of APP(695) Swedish (K595N/M596L) (APP(sw)) by transient cotransfection of HEK 293 cells with: 1) X11alpha (X11alpha-wt, N-MID-CID-PTB-PDZ-PDZ-C), 2) amino-terminal deletion (X11alpha-DeltaN, PTB-PDZ-PDZ), 3) carboxyl-terminal deletion (X11alpha-DeltaPDZ, MID-CID-PTB), or 4) deletion of both termini (PTB domain only, PTB). The carboxyl terminus of X11alpha was required for stabilization of APP(sw) in cells. In contrast, the amino terminus of X11alpha was required to stimulate APPs secretion. X11alpha, X11alpha-DeltaN, and X11alpha-PTB, but not X11alpha DeltaPDZ, were effective inhibitors of Abeta40 and Abeta42 secretion. These results suggest that additional protein interaction domains of X11alpha modulate various aspects of APP metabolism. PMID- 11010979 TI - Lead poisoning. PMID- 11010980 TI - Dysfunctional voiding. PMID- 11010981 TI - Back to basics: host responses to infection. PMID- 11010982 TI - Earning CME credit-completing the PIR quiz PMID- 11010983 TI - The child and death. PMID- 11010985 TI - Treatment of antibody deficiency syndromes. PMID- 11010986 TI - Message from the Incoming Editor. PMID- 11010987 TI - A Tribute to Paul Green. PMID- 11010988 TI - New Evidence for the Role of Mechanical Forces in the Shoot Apical Meristem. AB - The mechanism for initiation of lateral organs in the shoot apical meristem is still unknown. In this article we investigate one critical component of a buckling mechanism of organ initiation (that is, the presence and distribution of compressive stresses in the meristem). Direct evidence for compression in the sunflower capitulum was obtained from the gaping pattern of shallow cuts and the propagation of fractures. Cuts gaped widely in the central region of the capitulum but remained closed, or nearly so, in the generative and differentiation regions, suggesting the presence of circumferential compression at these locations. Fractures were initiated in the generative region and propagated circumferentially over most of their length. They did not cross the generative region perpendicularly, suggesting again the presence of compressive stresses in the circumferential direction. This conclusion was confirmed by the stress distribution computed from the geometry of the capitulum at three stages of development. One interpretation of these results is that the generative region corresponds to a zone of compression that could control the initiation of new primordia by means of buckling of the tunica layer. PMID- 11010989 TI - Leaves as Shell Structures: Double Curvature, Auto-Stresses, and Minimal Mechanical Energy Constraints on Leaf Rolling in Grasses. AB - Grass leaves are natural examples of shell structures because they are thin and display a double curvature. An important mechanical property of shells is that changes in longitudinal and transverse curvatures are not independent. The basis of this mechanical coupling is presented using simple diagrams. The relevance of the structural constraints for the processes of hydronastic rolling and developmental unrolling in grass leaves is then reviewed. I show that mechanical constraints can explain a large part of the genetic and developmental variability of hydronastic rolling in grasses, without reference to specific anatomic features such as bulliform cells. Mechanical analysis of a rolled maize mutant also revealed that developmental unrolling is not limited to a pure transverse expansion of hinge cells and involves both longitudinal and transverse dimensional changes in the upper epidermis. Interest in using mechanical models as a tool to reveal structural interactions at the tissue and organ level is discussed, and the importance of Paul Green's biophysical approach to the study of plant morphogenesis is emphasized. PMID- 11010990 TI - Tensile Tissue Stress Affects the Orientation of Cortical Microtubules in the Epidermis of Sunflower Hypocotyl. AB - In turgid multicellular organs, it is convenient to differentiate between the two kinds of tensile forces acting in cell walls as a result of turgor pressure. The primary forces occur both in situ and in cells isolated from the organ, whereas the secondary forces occur only in situ. The latter are an unavoidable physical consequence of the variation in mechanical parameters of tissues forming layers or strands. The most rigid tissue is under maximal tensile force, whereas the least rigid is under maximal compressive force. These forces cause tissue stresses (that is, certain tissues are under tensile stress, whereas others are under compressive stress in the organ). The primary and secondary forces result in primary and secondary stress in cell walls, respectively. The anisotropy of the primary stress is a function of cell shape. For instance, in cylindric cells the anisotropy expressed as the ratio of longitudinal to transverse stresses is 0.5. The anisotropy of the secondary stress is a function of the compound structure of the organ. For example, in the epidermis of sunflower hypocotyl, the longitudinal secondary stress is much higher than the transverse stress. The primary and secondary stresses are superimposed, and, as a consequence, the stress anisotropy in the outer thick walls of epidermal cells is greater than 1. These outer epidermal walls transmit most of the tissue stress. When the epidermis is peeled but remains turgid, only primary stress remains, but loading of the peel can reestablish the original stress anisotropy. We studied the effect of stress anisotropy changes on the orientation of cortical microtubules (CMTs) in the sunflower hypocotyl epidermis. We showed that changes in stress anisotropy cause the CMT orientation to change in the direction of maximal wall stress. In situ, the relatively high tensile tissue stress in the epidermis causes maximal stress in the longitudinal direction and relatively steep CMT orientation. When the tissue stress is removed from the epidermis by peeling, the CMTs tend to reorient toward the transverse direction, which is the direction of maximal stress in the primary component. On application of external longitudinal stress, to substitute for tissue stress, CMTs tend to reorient in the longitudinal direction. However, a relatively high rate of plastic strain is caused by the stress applied to the peel in an acid medium. This produces a less steep orientation of CMTs. It appears that the change in stress anisotropy orients the CMT in the direction in which the stress is maximal after the change, but there is also some effect of the growth rate on the orientation. PMID- 11010991 TI - Co-Ordination of Cell Division and Tissue Expansion in Sunflower, Tobacco, and Pea Leaves: Dependence or Independence of Both Processes? AB - Temporal analyses of cell division and tissue expansion in pea, tobacco, and sunflower leaves reveal that both processes follow similar patterns during leaf development. Relative cell division and relative tissue expansion rates are maximal and constant during early leaf development, but they decline later. In contrast, relative cell expansion rate follows a bell-shaped curve during leaf growth. Cell division and tissue expansion have common responses to temperature, intercepted radiation, and water deficit. As a consequence, final leaf area and cell number remain highly correlated throughout a large range of environmental conditions for these different plant species, indicating that cell division and tissue expansion are co-ordinated during leaf development. This co-ordination between processes has long been explained by dependence between both processes. Most studies on dicotyledonous leaf development indicate that leaf expansion rate depends on the number of cells in the leaf. We tested this hypothesis with a large range of environmental conditions and different plant species. Accordingly, we found a strong correlation between both absolute leaf expansion rate and leaf cell number. However, we showed that this relationship is not necessarily causal because it can be simulated by the hypothesis of independence between cell division and tissue expansion according to Green's theory of growth (1976). PMID- 11010992 TI - Clonal Analysis Provides Evidence for Transient Initial Cells in Shoot Apical Meristems of Seed Plants. AB - Drift of mutated sectors in sectorial or mericlinal plant chimeras has been interpreted as indirect evidence of initial impermanence at the apex. However, the same effect may result from mutation in noninitial cells positioned close to the vertex of the apical dome. Clonal analysis of the cell packets present in the superficial layer of spruce and magnolia apices provided the library of patterns suggesting that the position and the number of initial cells, and in some cases also the meristem axis inclination, may change over time. Multicellular clones originating from a single cell have been found in the geometric center of some apices, whereas in other apices the cellular center (where three or four clonal borders meet) did not correspond to the geome PMID- 11010993 TI - Effect of Soil pH on Growth and Cation Deposition in the Root Tip of Zea mays L. AB - The effects of sandy soil pH on the distribution of growth velocities and on cation concentrations and deposition rates in root growth zones of Zea mays L. seedlings were investigated. The pH values of the rooting medium varied between 4.2 and 8.6 in sand culture (70% saturated) without external supply of nutrients. At all pH values, densities (in umoles per g fresh weight) of potassium, magnesium, and calcium increased toward the root tip. Lower pH in the medium increased calcium tissue density fivefold and magnesium density 1.7-fold, whereas the density of potassium, the overall elongation rate, and the growth velocity distribution did not show any significant pH dependence. Throughout the growth zone the deposition rates of the divalent cations, as calculated on the basis of the continuity equation, increased with lower pH. The data are consistent with the hypothesis that the effects of pH on the cation deposition rates are due to the increase in the divalent cation concentration of the soil solution at low pH and that the abundant uronic acid residues of the young walls of the meristem provide a reservoir of storage capacity for Ca and Mg under conditions of low nutrient availability. PMID- 11010994 TI - Comparison of Leaf Plastochron Index and Allometric Analyses of Tooth Development in Arabidopsis thaliana. AB - Two methods of analyses were used to investigate tooth development in serrate (se) mutant and wild-type Columbia-1 (Col-1) Arabidopsis thaliana leaves. There were almost twice as many teeth with deeper sinuses and two orders of toothing on the margins of serrate compared with Columbia-1 leaves. The main objective of this study was to test three hypotheses relative to the source of polymorphism in tooth development: (i) Teeth share similar growth rates and initial sizes, but the deeper teeth are initiated earlier in leaf development. (ii) Teeth share similar timing of initiation and growth rates, but the deeper teeth have a larger initial size. (iii) Teeth share similar timing of initiation and initial sizes, but the deeper teeth have a faster growth rate. Leaf plastochron index (LPI) was used as the time variable for leaf development. Results showed teeth in se were initiated at -27 LPI, 15 plastochrons earlier than those of Col-1. Serrate leaf expansion was biphasic, with the early phase expanding at half the relative plastochron rate of the later phase, which equaled the constant relative expansion rate of Col-1 leaves. Allometric analyses of tooth development obscured the interactions between time of tooth and leaf initiation and the early phase of leaf expansion characteristic of serrate leaves and teeth. Timing of developmental events that allometric analysis obscured can be readily detected with the LPI as a developmental index. PMID- 11010995 TI - Ball tonometry: a rapid, nondestructive method for measuring cell turgor pressure in thin-walled plant cells. AB - In this article we describe a new method for the determination of turgor pressures in living plant cells. Based on the treatment of growing plant cells as thin-walled pressure vessels, we find that pressures can be accurately determined by observing and measuring the area of the contact patch formed when a spherical glass probe is lowered onto the cell surface with a known force. Within the limits we have described, we can show that the load (determined by precalibration of the device) divided by the projected area of the contact patch (determined by video microscopy) provides a direct, rapid, and accurate measure of the internal turgor pressure of the cell. We demonstrate, by parallel measurements with the pressure probe, that our method yields pressure data that are consistent with those from the pressure probe. Also, by incubating target tissues in stepped concentrations of mannitol to incrementally reduce the turgor pressure, we show that the pressures measured by tonometry accurately reflect the predicted changes from the osmotic potential of the bathing medium. The advantages of this new method over the pressure probe are considerable, however, in that we can move rapidly from cell to cell, taking measurements every 20 s. In addition, the nondestructive nature of the method means that we can return to the same cell repeatedly for periodic pressure measurements. The limitations of the method lie in the fact that it is suitable only for superficial cells that are directly accessible to the probe and to cells that are relatively thin walled and not heavily decorated with surface features. It is also not suitable for measuring pressures in flaccid cells. PMID- 11010996 TI - The Double-Water-Film Electrode: A Device for Measuring the Resistance and the Capacitance of the Internode/Node Interface of Chara as Functions of Time and Temperature. AB - A "double-water-film electrode technique" has been developed for the long-term characterization of the electrical properties across the interface between the nodal (N) and internodal (A or B) cells and the vacuole along the length of an internode of Chara as a function of time and temperature. The electrode unit consisted of a pair of the water-film electrodes described elsewhere (Chilcott 1988; Chilcott and others 1983; Coster and others 1984; Lucas 1985; and Ogata 1983). The distance between two water-film probes was fixed at 1.0 cm. By scanning the electrode unit, the spatial variations in electrical resistance and capacitance along the longitudinal axis of Chara were observed. Analysis was performed by applying an electrical equivalent circuit for the biomembrane (Philippson 1921). Across the internode (-A or -B)/central nodal cells interface, the specific parallel resistance (Rm) and the parallel capacitance (Cm) at 20 degrees C were 30 +/- 5 x 10(-3) Omegam(2) and 1.5 +/- 0.5 x 10(-1)Fm(-2) (at 30 Hz), respectively. And the series resistance, corresponding to the vacuole of the internode was 8 x 10(-3) Omegam(2). Study of temperature dependencies of Rm and Cm suggested that a dynamic homeostatic regulation was operating at the interface where numerous plasmodesmata were observed with an electron microscope (Pickett Heaps 1967; Spanswick and Costerton 1967). Assuming that the individual cylinder of plasmodesma was filled only with cytoplasm, the number of plasmodesma per interface was estimated at 2.6 x 10(5). PMID- 11010997 TI - Effect of Abscisic Acid on Banana Fruit Ripening in Relation to the Role of Ethylene. AB - The role of abscisic acid (ABA) in banana fruit ripening was examined with the ethylene binding inhibitor, 1-methylcyclopropene (1-MCP). ABA (0, 10(-5), 10(-4), or 10(-3) mol/L) was applied by vacuum infiltration into fruit. 1-MCP (1 uL/L) was applied by injecting a measured volume of stock gas into sealed glass jars containing fruit. Fruit ripening, as judged by ethylene evolution and respiration associated with color change and softening, was accelerated by 10(-4) or 10(-3) mol/L ABA. ABA at 10(-5) mol/L had no effect. The acceleration of ripening by ABA was greater at 10(-3) mol/L than at 10(-4) mol/L. ABA-induced acceleration of banana fruit ripening was not observed in 1-MCP treated fruit, especially when ABA was applied after exposure to 1-MCP. Thus, ABA's promotion of ripening in intact banana fruit is at least partially mediated by ethylene. Exposure of ABA treated fruit to 0.1 uL/L ethylene for 24 h resulted in increased ethylene production and respiration, and associated skin color change and fruit softening. Control fruit (no ABA) was unresponsive to similar ethylene treatments. The data suggest that ABA facilitates initiation and progress in the sequence of ethylene mediated ripening events, possibly by enhancing the sensitivity to ethylene. PMID- 11010998 TI - Opiate receptor avidity is increased in rhesus monkeys following unilateral optic tract lesion combined with transections of corpus callosum and hippocampal and anterior commissures. AB - Opiate receptor avidity (B(max)'/K(D)) was measured in four rhesus monkeys following unilateral lesioning of the optic tract combined with transection of the corpus callosum and the hippocampal and anterior commissures depriving one hemisphere of visual input (Tract and Split), two animals with transection of commissures only (Split), and nine healthy monkeys with positron emission tomography (PET) and 6-deoxy-6-beta-[(18)F]fluoronaltrexone (cyclofoxy, CF), a mu and kappa-opiate receptor antagonist. Opiate receptor avidity was found to be significantly higher in the Tract and Split animals, only, bilaterally, throughout the lateral cortex and in the cingulate and posterior putamen (41 117%). Ipsilateral changes were consistently greater than those contralateral, but this asymmetry was of statistical significance only in the parietal and occipital cortices. Cyclofoxy avidity was decreased in the medial cortex of both the Tract and Split and Split animals ( approximately 25%). The results suggest that opiate pathways undergo extensive alteration in response to changes in brain functional activities brought about through hemispheric visual deprivation. PMID- 11010999 TI - Release of synaptic zinc is substantially depressed by conventional brain slice preparations. AB - Research on synaptically-released zinc is frequently done in vitro with acute brain slice preparations. We show here the in vitro hippocampal slice preparation has two major pitfalls for zinc research. First, up to 50% of the synaptic zinc is lost during slice cutting and/or the first 10 min of slice incubation, with the losses being most pronounced on the edges of the slice. Second, the release of the remaining zinc from a slice is substantially depressed (up to 50%) at the low temperatures (32 degrees C) typically used for brain slice studies. In concert, these effects reduce zinc release about 75% in vitro, compared to in vivo. Implications for research on synaptically-released zinc are discussed. PMID- 11011000 TI - Bilirubin does not modulate ionotropic glutamate receptors or glutamate transporters. AB - Bilirubin, a product of haemoglobin metabolism, has been suggested to damage neurons by increasing activation of N-methyl-D-aspartate (NMDA) receptors when it reaches high levels in the blood [15,19], as occurs in neonatal jaundice [7]. Bilirubin is also generated in the brain following synthesis of the messenger carbon monoxide (CO) by haem oxygenase, and haem oxygenase is upregulated in Alzheimer's disease [23]. We examined the effect of bilirubin on currents generated by NMDA and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors in hippocampal pyramidal cells, and on glutamate transporter currents in retinal glial cells. Bilirubin did not modulate either receptor-gated currents or transporter currents. These data show the negative, but important result that bilirubin does not induce neuronal death by acting directly on NMDA or AMPA receptors, nor indirectly by blocking glutamate uptake and raising the extracellular concentration of glutamate. PMID- 11011001 TI - A comparison of morphine analgesic tolerance in male and female mice. AB - Studies comparing morphine tolerance in males and females are rare, and all studies to date have utilized the rat. To generalize from findings with rats morphine tolerance was investigated in male and female mice using the tail withdrawal test. Three and 7 days of systemic morphine injections produced significant but unequal rightward shifts in the morphine dose-response curve such that females displayed greater increases in analgesic ED(50) values when compared to males. In a separate experiment, males and females displayed similar reductions in morphine analgesic sensitivity when %MPE (maximum possible effect) and %total (area under the curve) were compared after 3 days of morphine. Differences in initial morphine sensitivity between sexes were not observed in either study. The data demonstrate that, in contrast to rats, female mice undergo greater reductions in morphine analgesia relative to males following chronic morphine, but this sex difference may depend on the method of assessing analgesia. Furthermore, the duration and/or cumulative dose of morphine treatment does not affect the expression of sex differences in morphine tolerance. PMID- 11011002 TI - Intracerebroventricular met-enkephalin administration modulates adjuvant arthritis. AB - The purpose of this study was to investigate the anti-inflammatory properties of intracerebroventricular met-enkephalin (met-enk) administration in an animal model of arthritis. Adjuvant arthritis was induced in rats by intradermal inoculation of mycobacterium butyricum and the effects of intraventricular met enk+thiorphan (enkephalinase inhibitor) were studied. Treatment was initiated either simultaneously with the bacterial inoculation (preventive group) or on post-inoculation day 17 after the appearance of inflammation (treatment group). The degree of inflammation was evaluated by measuring the diameter and the circumference of the ankle joint immediately before the sacrifice (day 31) and by histologic examination of ankle joint sections. The results of this study revealed that combined intraventricular injections of met-enk+thiorphan reduced the arthritic-like inflammation in the preventive group as well as in the treatment group. These findings suggest that centrally applied met-enk+thiorphan may suppress the development adjuvant arthritis as well as the symptoms of manifest arthritis. Thus central met-enk may be involved in both hypothalamic pituitary adrenal axis and immune forms of stress-induced modulation. PMID- 11011003 TI - The role of intrinsic and agonist-activated conductances in determining the firing patterns of preoptic area neurons in the guinea pig. AB - Whole-cell and intracellular recordings were made in coronal hypothalamic slices prepared from ovariectomized female guinea pigs. 62% of preoptic area (POA) neurons fired action potentials in a bursting manner, and exhibited a significantly greater afterhyperpolarization (AHP) than did non-bursting POA neurons. The majority (70%) of POA neurons (n=76) displayed a time-dependent inward rectification (I(h)) that was blocked by CsCl (3 mM) or by ZD 7288 (30 microM). In addition, 51% of the cells expressed a low-threshold spike (LTS) associated with a transient inward current (I(T)) that was blocked by NiCl(2) (200 microM). A smaller percentage of POA neurons (29%) expressed a transient outward, A-type K(+) current that was antagonized by a high concentration of 4 aminopyridine (3 mM). Moreover, POA neurons responded to bath application of the mu-opioid receptor agonist DAMGO (93%) or the GABA(B) receptor agonist baclofen (83%) with a membrane hyperpolarization or an outward current. These responses were accompanied by a decrease in input resistance or an increase in conductance, respectively, and were attenuated by BaCl(2) (100 microM). In addition, the reversal potential for these responses closely approximated the Nernst equilibrium potential for K(+). These results suggest that POA neurons endogenously express to varying degrees an AHP, an I(h), an I(T) and an A-type K(+) current. The vast majority of these neurons also are inhibited upon mu opioid or GABA(B) receptor stimulation via the activation of an inwardly rectifying K(+) conductance. Such intrinsic and transmitter-activated conductances likely serve as important determinants of the firing patterns of POA neurons. PMID- 11011004 TI - Neurotoxicity of HIV-1 proteins gp120 and Tat in the rat striatum. AB - HIV-associated dementia complex is a serious disabling disease characterized by cognitive, behavioral and motor dysfunction. Basal ganglia involvement in HIV-1 infection may be responsible for some of the psychomotor symptoms associated with HIV dementia. The objectives of the present study were to determine: (1) whether gp120 and Tat produce striatal toxicity, and (2) whether gp120 and Tat show synergistic toxicity in the striatum. In these studies, the recombinant proteins gp120, Tat, or saline (0.9%) were stereotaxically injected in the striatum of adult male rats. The striatal sections were evaluated for area of tissue loss (Cresyl-violet stained sections) and the number of GFAP immunoreactive cells 7 days after the injections. Doses of gp120 250 ng/microl or higher and Tat 5 microg/microl or higher produced a significant area of tissue loss and significantly increased the number of GFAP reactive cells. We found no toxicity in animals treated with immunoabsorbed gp120 or Tat. Combined gp120 (100 ng/microl)+Tat (1 microg/microl) injections into the rat striatum significantly increased the area of tissue loss and altered morphology and increased number of GFAP reactive cells, as compared to controls. Thus, the present results suggest the involvement of gp120 and Tat in striatal toxicity and provide a model for further studies to fully characterize their role in HIV-1 toxicity and to develop therapeutic strategies for HIV-1 associated dementia complex. PMID- 11011005 TI - Diadenosine polyphosphates facilitate the evoked release of acetylcholine from rat hippocampal nerve terminals. AB - Diadenosine polyphosphates are present in synaptic vesicles, are released upon nerve stimulation and possess membrane receptors, namely in presynaptic terminals. However, the role of diadenosine polyphosphates to control neurotransmitter release in the CNS is not known. We now show that diadenosine pentaphosphate (Ap(5)A, 3-100 microM) facilitated in a concentration dependent manner the evoked release of acetylcholine from hippocampal nerve terminals, with a maximal facilitatory effect of 116% obtained with 30 microM Ap(5)A. The selective diadenosine polyphosphate receptor antagonist, diinosine pentaphosphate (Ip(5)I, 1 microM), inhibited by 75% the facilitatory effect of Ap(5)A (30 microM), whereas the P(2) receptor antagonists, suramin (100 microM) and pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS, 10 microM) only caused a 18-24% inhibition, the adenosine A(1) receptor antagonist, 1,3-dipropyl 8-cyclopentylxanthine (20 nM), caused a 36% inhibition and the adenosine A(2A) receptor antagonist, 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo [2,3-a][1,3, 5]triazin-5-ylamino]ethyl)phenol (ZM 241385, 20 nM), was devoid of effect. These results show that diadenosine polyphosphates act as neuromodulators in the CNS, facilitating the evoked release of acetylcholine mainly through activation of diadenosine polyphosphate receptors. PMID- 11011006 TI - Short-term ethanol exposure alters calbindin D28k and glial fibrillary acidic protein immunoreactivity in hippocampus of mice. AB - The effects of a short-term ethanol treatment on hippocampus have been studied in mice exhibiting intoxication signs. The alterations of neurons and astrocytes as well as quantitative changes of calbindin D28k-immunoreactivity and glial fibrillary acidic protein-immunoreactivity (GFAP-IR) in selected regions of the dorsal hippocampus were examined using anti-calbindin and anti-GFAP monoclonal anti-body (mAb), respectively. The administration of 6% (v/v) ethanol during first week led to the neuronal death and decrease of the total number of calbindin-IR neurons in the examined brain regions. Moreover, the calbindin positive neurons were shown to have diminished processes following short-term ethanol exposure. These neuronal changes were associated with the increase of the GFAP-IR astrocytes. Hypertrophy of cell bodies and cytoplasmic processes of reactive astrocytes were also seen. In addition, dense, thick and highly-stained GFAP-IR cells with long processes in granular cell layer appeared entering into molecular layer of dentate gyrus. In agreement with the discrepancy percentage of neuronal cell loss and increase of reactive astrocytes detected by calbindin and GFAP-IR using image quantitative analysis, the regional differences in the vulnerability to the neurotoxic effects following short-term ethanol exposure were found: CA3>CA2>CA1>DG. These findings also illustrate the importance of correlation between calbindin and GFAP-IR when determining the morphological alteration of neuron and astroglial following short-term ethanol treatment. The disruption of calbindin and GFAP could affect neuronal-astroglial interaction, resulting in disturbance of behaviors dependent on hippocampus. PMID- 11011007 TI - Effects of threonine injections in the lateral hypothalamus on intake of amino acid imbalanced diets in rats. AB - Previous work from this laboratory suggests that animals decrease their intake of an amino acid imbalanced diet (IMB), due in part to a drop in the concentration of the dietary limiting amino (DLAA) in the anterior piriform cortex (APC). Administration of the DLAA, but not of a non-limiting amino acid into the APC, blocks the anorectic response to IMB. To our knowledge, the effects of DLAA injections on intake of a diet devoid of the DLAA (DEV), have not been examined in areas outside the APC. We hypothesized that the LH is a potential chemosensory area for DLAA. Our objectives were: (1) to determine whether injections of the DLAA threonine into the lateral hypothalamus (LH) alter intake of a threonine devoid diet (DEV); and (2) to examine the dose-response effects of threonine injections into the LH on intake of threonine-corrected diet (COR). Administration of threonine into the LH stimulated DEV intake during the first 6 h at the 0.25 and 1-nmol doses by approximately 26 and 24%, respectively. Threonine (0.25, 2.5 nmol) did not alter COR intake at any time during the first 12 h. Our results suggest that: (1) the LH, along with the APC, likely acts as a chemosensory brain area for indispensable amino acids; and (2) both the APC and LH are part of a circuit that is involved in the short term anorectic response to amino acid imbalanced diets. PMID- 11011008 TI - Odor input generates approximately 1.5 Hz and approximately 3 Hz spectral peaks in the helix pedal ganglion. AB - In 1999 we reported that odorants evoke in the Helix pedal ganglion (PG) activity patterns which are largely odorant-specific and related to the nature of odor and its behavioral output. Notably, some activities (for example, approximately 1.5 and approximately 3 Hz), nonspecific to odorants, were consistently evoked in PG. The present contribution goes farther in a deeper survey of the intrinsic and odorant-evoked activities of PG with special weight on the nonspecific fluctuations. We address the following questions. (i) What are the features of the activities? (ii) Are they comparable to the activities found in the motor systems of the other invertebrates? (iii) To what functions can they be related? Three main frequency components represented by power peaks at <1 Hz, 1-2 Hz and 2 8 Hz seem to feature the response activities of PG. (a) The aversive odorants induce odorant-specific patterns represented by peak power frequencies at <1 Hz. (b) The oscillation at approximately 1 Hz, which exists intrinsically in the Helix PG, can be specifically enhanced by appetitive odors. Activities induced in the procerebrum (PC), the visceral ganglion (VG) and PG by appetitive odorants, such as ethanol and apple, peak at 1.3-2 Hz, whereas those induced by aversive ones, such as formic acid and onion at <1 Hz. (c) The 2-8 Hz components always accompany the odorant-evoked activities of the PG either as the second or third strongest component, or in the form of conspicuous, long-lasting approximately 3 Hz oscillations. (d) The nonspecific odor-evoked 1-2 Hz and approximately 3 Hz activities, and the intrinsic approximately 1 Hz activity of the PG seem to be interrelated by a degree of mutual exclusion. We may therefore consider these activities as elementary, slow components that are involved in the processing of signals in this ganglion. It can be inferred from the findings in other invertebrates that the 1-3 Hz spontaneous discharge is strongly connected with motor activity that involves the feedback mechanism of the procerebro-cerebro buccal or -procerebro-cerebro-pedal circuit. Our approach differs from most others reported so far in the following aspects: (i) use of gross steel electrodes for recording population activities; (ii) lengthy stimulation (10 min); (iii) long observation during and after stimulation; (iv) power spectral presentation of temporal evolution of activity patterns; (v) estimation of peak power frequency by Frequency-Amplitude Plot (FAP) (obtained from signals averaged in the frequency domain; a method based on systems theory). PMID- 11011009 TI - Quisqualate induces an inward current via mGluR activation in neocortical pyramidal neurons. AB - Activation of metabotropic glutamate receptors (mGluRs) has multiple effects on the excitability of pyramidal neurons in rat frontal neocortex. Synaptic transmission and intrinsic excitability are both affected. During studies of the effects of quisqualate on synaptic activity, it was observed that quisqualate also induced a slow inward current. Whole-cell patch clamp recordings were obtained from layer II/III pyramidal neurons of neocortical slices in vitro. The bath solution contained APV, CNQX and bicuculline to block ionotropic glutamate and GABA(A) receptors. At a holding potential of -70 mV, quisqualate (2 microM) induced an inward current of about 60 pA. The response was reversible upon washing. This current was associated with an increase in membrane conductance and was still seen in the presence of TTX (0.5 microM). Bath application of the nonselective mGluR antagonist, (R, S)-alpha-methyl-4-carboxyphenyglycine (MCPG, 200-500 microM) reduced the current by 70%. Other mGluR agonists (ACPD, DHPG, L CCG-1 and L-AP4) did not induce a significant inward current at the concentrations tested. The current-voltage relation of the quisqualate-induced current was linear with a reversal potential near 0 mV suggesting involvement of nonselective cation channels. The quisqualate-induced inward current was markedly reduced (72%) with 200 microM GDP-beta-S in the pipette solution, indicating that it is a postsynaptic phenomenon mediated by a G-protein dependent mechanism. These results suggest that mGluRs can directly increase the postsynaptic excitability of pyramidal cells. PMID- 11011010 TI - Enhancement of the acoustic startle response by dopamine agonists after 6 hydroxydopamine lesions of the substantia nigra pars compacta: corresponding changes in c-Fos expression in the caudate-putamen. AB - Rats with 6-hydroxydopamine (6-OHDA) lesions of the nigrostriatal pathway show enhanced locomotor and stereotyped behaviors when challenged with direct and indirect dopamine (DA) agonists due to the development of postsynaptic supersensitivity. To determine if this phenomenon generalizes to other motor behaviors, we have used this rat model of Parkinson's disease to examine the effects of the direct dopamine D(1) receptor agonist SKF 82958 and the indirect DA agonist L-3,4-dihydroxyphenylalanine (L-DOPA) on the acoustic startle response. In addition, we used the expression of c-Fos protein as a marker of neuronal activity to assess any corresponding drug-induced changes in the caudate putamen (CPu) after L-DOPA administration. Male Sprague-Dawley rats received bilateral injections of 6-OHDA into the substantia nigra pars compacta and 1 week later were tested for startle after systemic administration of SKF 82958 (0.05 mg/kg) or L-DOPA (1, 5, 10 mg/kg). SKF 82958 produced a marked enhancement of startle with a rapid onset in 6-OHDA-lesioned but not SHAM animals. L-DOPA produced a dose- and time-dependent enhancement of startle in 6-OHDA-lesioned rats that had no effect in SHAM animals even at the highest dose (10 mg/kg). Furthermore, L-DOPA produced a dramatic induction of c-Fos in the CPu in 6-OHDA lesioned animals. Consistent with other literature, these data suggest that neurons in the CPu become supersensitive to the effects of DA agonists after 6 OHDA-induced denervation of the nigrostriatal pathway and that supersensitive dopamine D(1) receptors may mediate the enhancement of startle seen in the present study. PMID- 11011011 TI - Autonomic and cardiovascular reflex responses to central estrogen injection in ovariectomized female rats. AB - The role of estrogen in central autonomic nuclei was examined in ovariectomized female Sprague-Dawley rats supplemented daily for 7 days with either estrogen (5 microg/kg; sc) or saline (0.9%; sc). Animals were subsequently anaesthetized with sodium thiobutabarbital (Inactin; 100 mg/kg; ip) and instrumented to record blood pressure and heart rate. Efferent vagal parasympathetic (VPNA) and renal sympathetic (RSNA) nerve activities were recorded and used to assess baseline and reflexive changes in autonomic tone. The cardiac baroreflex was evoked using a single bolus injection of phenylephrine (0.1 mg/kg) both before and following either intrathecal injection of estrogen (0.5 microM; 1 microl) or bilateral injection of estrogen (0.5 microM; 100 nl/side) into several central autonomic nuclei. In estrogen-replaced rats, both the baseline and PE-evoked values for mean arterial pressure and RSNA were significantly decreased following injection of estrogen into the nucleus tractus solitarius (NTS), rostral ventrolateral medulla (RVLM), parabrachial nucleus (PBN), central nucleus of the amygdala (CNA) and the intrathecal space. Baseline heart rate and VPNA were significantly decreased following injection of estrogen into NTS, nucleus ambiguous (Amb), PBN and the intrathecal space. PE-evoked changes in heart rate and VPNA were significantly enhanced following injection of estrogen into these same nuclei. Injection of estrogen into the insular cortex (IC) produced significant decreases in baseline and PE-evoked RSNA only. The cardiac baroreflex was significantly enhanced following injection of estrogen into all nuclei and the intrathecal space. In saline-replaced females, injection of estrogen into NTS, RVLM, Amb and the intrathecal space had similar effects on both baseline and PE-evoked parameters although of a reduced magnitude compared to estrogen-replaced rats. However, no significant changes in autonomic tone and baroreflex function were observed following the injection of estrogen into the PBN, CNA or IC of saline replaced rats. These results demonstrate a role for estrogen in central autonomic nuclei in female rats and suggest a possible alteration of estrogen receptor distribution or efficacy within the central nervous system of estrogen-deficient female rats. PMID- 11011013 TI - Haemodynamic correlates of penumbral depolarization following focal cerebral ischaemia. AB - Transient ischaemic depolarizations (IDs) are thought to play a key role in the pathogenesis of focal cerebral ischaemia. Most transient IDs are akin to spreading depression (SD), although a negative DC shift is not observed in half the cases. The other IDs may represent transient anoxic depolarizations. Using cortical DC and blood flow recordings, following middle cerebral artery occlusion in rats, we show here that: (i) these later depolarizations do indeed represent transient anoxic depolarizations; (ii) SD-like IDs, DC and haemodynamic parameters are similar to those of SDs when blood flow remains close to base line and; (iii) when blood flow decreases, the hyperaemia associated with SD-like IDs is largely reduced and there is an increasing proportion of cortical sites which fail to display a DC shift. These data demonstrate the coexistence of two mechanisms of IDs, and yield new information as to the flow-dependence of DC and haemodynamic correlates of SD-like IDs, the pathophysiological significance of which remains to be determined. PMID- 11011012 TI - The 4F2hc/LAT1 complex transports L-DOPA across the blood-brain barrier. AB - L-DOPA is transported across the blood-brain barrier (BBB) by an amino acid transporter, system L. Recently, it has been demonstrated that system L consists of two subunits, 4F2hc and either LAT1 or LAT2. 4F2hc/LAT1 and 4F2hc/LAT2 show different transport characteristics, while their distribution in the brain has not been determined. To clarify whether 4F2hc/LAT1 participates in L-DOPA transport across the BBB, we first examined the expression of 4F2hc/LAT1 in the mouse brain capillary endothelial cell line, MBEC4, as an in vitro BBB model. Northern hybridization and immunoblotting revealed that both 4F2hc and LAT1 are expressed and form a heterodimer in MBEC4 cells. To confirm whether 4F2hc/LAT1 acts as system L to transport L-DOPA, we characterized L-DOPA uptake into the cells. The uptake process was time-dependent, temperature-sensitive, and Na(+) independent. Neutral amino acids with bulky side chains and a bicyclic amino acid, 2-aminobicyclo-[2, 2,1]-heptane-2-carboxylic acid (BCH), inhibited L-DOPA uptake into MBEC4 cells to a great extent, while an acidic amino acid, basic amino acids, and glycine had no effect. Other neutral amino acids, such as alanine, asparagine, glutamine, serine, and threonine inhibited L-DOPA uptake by 40-70% at most. These characteristics are more compatible with those of 4F2hc/LAT1, rather than those of 4F2hc/LAT2. Finally, immunohistochemistry with anti-LAT1 antibody demonstrated that LAT1 is predominantly expressed in the microvessels of the central nervous system. This is the first report showing that the 4F2hc/LAT1 complex participates in L-DOPA transport across the BBB. PMID- 11011014 TI - Effects of amygdala lesions on sleep in rhesus monkeys. AB - The amygdala is important in processing emotion and in the acquisition and expression of fear and anxiety. It also appears to be involved in the regulation of sleep and wakefulness. The purpose of this study was to assess the effects of, fiber-sparing lesions of the amygdala on sleep in rhesus monkeys (Macaca mulatta). We recorded sleep from 18 age-matched male rhesus monkeys, 11 of which had previously received ibotenic acid lesions of the amygdala and seven of which were normal controls. Surface electrodes for sleep recording were attached and the subjects were seated in a restraint chair (to which they had been adapted) for the nocturnal sleep period. Despite adaptation, control animals had sleep patterns characterized by frequent arousals. Sleep was least disrupted in animals with large bilateral lesions of the amygdala. They had more sleep and a higher proportion of rapid-eye-movement (REM) sleep than did either animals with smaller lesions or control animals. Based on these results, it seems likely that, in the primate, the amygdala plays a role in sleep regulation and may be important in mediating the effects of emotions/stress on sleep. These findings may also be relevant to understanding sleep disturbances associated with psychopathology. PMID- 11011015 TI - Ovarian steroids influence the activity of neuroendocrine dopaminergic neurons. AB - The secretion of prolactin (PRL) from the anterior lobe (AL) of the pituitary gland is tonically inhibited by dopamine (DA) of hypothalamic origin. While ovarian steroids play a role in the regulation of the secretion of PRL, their effect on all three populations of hypothalamic neuroendocrine dopaminergic neurons is not fully understood. In this study we describe the effects of ovarian steroids on regulation of the release of DA from tuberoinfundibular dopaminergic (TIDA), tuberohypophyseal dopaminergic (THDA) and periventricular-hypophyseal dopaminergic (PHDA) neurons. Adult female rats were bilaterally ovariectomized (OVX) and, 10 days following ovariectomy (day 0), injected with corn oil (vehicle), estrogen, or estrogen plus progesterone (day 1). Animals were sacrificed every 2 h from 09.00 to 21.00 h by rapid decapitation. Trunk blood was collected and the concentration of PRL in serum was determined by radioimmunoassay. The median eminence (ME) and the AL, intermediate (IL) and neural (NL) lobes of the pituitary gland were dissected and the concentration of DA and DOPAC in each was measured by HPLC-EC. OVX rats presented small but significant increases in the secretion of PRL at 15.00 and 17.00 h. Replacement of estrogen or estrogen plus progesterone increased the basal concentration of PRL. Moreover, injection of estrogen only, or estrogen plus progesterone increased the concentration of PRL in serum at 15.00 h through 19.00 h, respectively, followed by a decrease to baseline thereafter. The turnover of DA in the ME and NL of OVX rats increased at 13.00 and returned to low levels. Turnover of DA in the IL of OVX rats increased in the morning by 11.00 h and remained elevated before decreasing by 17.00 h. The turnover of DA in the ME, IL and NL of OVX rats increased by 19.00 h. Injection of estrogen advanced the increase of TIDA activity by 2 h in the ME compared to OVX rats. Moreover, administration of estrogen suppressed the activity of THDA and PHDA neurons in the afternoon compared to OVX rats. In estrogen plus progesterone-treated rats, the activity of hypothalamic neuroendocrine dopaminergic neurons terminating in the ME, IL, and NL was inhibited prior to the increase in the secretion of PRL. The concentration of DA in the AL diminished prior to the estrogen-induced increase of PRL. Administration of progesterone, in concert with estrogen, delayed the increase of PRL in serum and the decrease of DA in the AL, compared to estrogen-treated rats, by 4 h. These data suggest a major role for ovarian steroids in controlling increases in the secretion of PRL by not only stimulating PRL release from lactotrophs, but also by inhibiting the activity of all three populations of hypothalamic neuroendocrine DAergic neurons. PMID- 11011016 TI - The effects of L-NAME on vestibular compensation and NOS activity in the vestibular nucleus, cerebellum and cortex of the guinea pig. AB - Nitric oxide (NO) has been implicated in the processes by which animals recover from peripheral vestibular damage ('vestibular compensation'). However, few data exist on the dose-response effects of systemic administration of the nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), on the vestibular compensation process. The aim of this study was to investigate the effects on compensation of 5, 10, 50 or 100 mM L-NAME administered by s.c osmotic minipump for 50 h following unilateral vestibular deafferentation (UVD) in guinea pig, either commencing the drug treatment at 4 h pre-UVD or at the time of the UVD (i.e., post-UVD). Post-UVD treatment with L-NAME, at any of the four concentrations used, had no effect on the compensation of spontaneous nystagmus (SN), yaw head tilt (YHT) or roll head tilt (RHT). By contrast, pre-UVD treatment with 100 mM L-NAME resulted in a significant decrease in SN frequency (P<0.05) and a change in the rate of its compensation (P<0.0005). Pre-UVD L-NAME resulted in a significant increase in the overall magnitude of YHT (P<0.005); however, post-hoc comparisons revealed no significant differences between any specific L NAME and vehicle groups. Pre-UVD L-NAME had no effect on RHT at any concentration. Analysis of NOS activity in the pre-UVD L-NAME treatment groups at 50 h post-UVD showed that only 100 mM L-NAME resulted in a significant decrease in NOS activity in the contralateral medial vestibular nucleus (MVN)/prepositus hypoglossi (PH) (P<0.05) and that NOS activity in the ipsilateral MVN/PH was not significantly affected. However, NOS activity was significantly inhibited in the bilateral cerebellum and cortices for several concentrations of L-NAME. These results suggest that pre-UVD systemic administration of L-NAME can significantly increase the rate of SN compensation in guinea pig and that this effect is correlated with inhibition of NOS activity in several regions of the CNS. PMID- 11011017 TI - Behavioral training-induced c-Fos expression in the rat nucleus basalis of Meynert during aging. AB - The present study investigated the behavioral training-induced c-Fos expression in the nucleus basalis of Meynert (nbM) in differently aged rats. This study demonstrated that the c-Fos expression in nbM was significantly increased and the peak occurred at 2 h after dark-avoidance training. Although the increase of c Fos expression was also observed after pseudotraining, the number of Fos-like immunoreactive neurons in pseudotrained rats was significantly less than that in dark-avoidance trained rats at each time-point. This result suggested that c-Fos expression might be involved in learning and memory processes. In addition, all the pseudotraining-, training- and memory arousing-induced c-Fos expression was decreased with increasing age, and the decrease was more notable in trained and memory aroused rats. This suggested that the total number of nbM neurons and/or the sensitivity of nbM neurons to experimental manipulations, especially learning and memory performance, might reduce during aging. PMID- 11011018 TI - Cellular determinants of reduced adaptability of the aging brain: neurotransmitter utilization and cell signaling responses after MDMA lesions. AB - Senescence is accompanied by the loss of neurons and synapses, and the maintenance of function depends on adaptive change at the levels of synaptic activity and cellular responsiveness. In the current study, we administered the neurotoxin MDMA, to young and aged mice and assessed the effects on indices of neuronal activity and cell signaling mediated through adenylyl cyclase. Young mice given MDMA showed 80% depletion of dopamine in the caudate and 30% depletion in the cerebral cortex; measurements of dopamine turnover indicated a compensatory upregulation of the activity of the remaining neurons in the caudate but downregulation in the cerebral cortex. Serotonin levels were comparatively less affected but serotonin turnover was decreased significantly in both regions. At the level of cell signaling, the young mice showed heterologous upregulation of adenylyl cyclase activity and a consequent enhancement of responses mediated through neurotransmitter receptors. In aged mice, MDMA treatment produced the same degree of lesioning but substantially different changes in neuronal activity and cell signaling. In the cerebral cortex, dopamine turnover was increased, and serotonin turnover decreased, effects opposite in direction to those seen in young mice. In the aged group, MDMA elicited heterologous loss of adenylyl cyclase responses instead of displaying the supersensitivity that had been seen in the young group. The aging brain thus displays maladaptation to the loss of monoaminergic input, effects that may augment the functional impairment associated with neurodegenerative disorders or stroke. PMID- 11011019 TI - Androgen receptor-immunoreactivity in the forebrain of the Eastern Fence lizard (Sceloporus undulatus). AB - Androgen receptor (AR) distribution in the lizard forebrain and optic tectum was examined using PG21 immunohistochemistry. In the male Eastern Fence lizard, AR immunoreactive (-ir) nuclei were observed in the medial preoptic area, ventromedial and arcuate hypothalamic nuclei, periventricular hypothalamus, premammillary nucleus, bed nucleus of the stria terminalis, and ventral posterior amygdala. Punctate immunostaining of neuronal processes (axons and/or dendrites) was concentrated in the cortex, hypothalamus, and optic tectum. AR-ir nuclei in the female brain were confined to the ventral posterior amygdala and ventromedial hypothalamic nucleus. The AR distribution in the lizard brain is similar to that reported for other vertebrate classes. Sex differences in AR-immunoreactivity may contribute to sex-specific behaviors in the Eastern Fence lizard. PMID- 11011020 TI - Motor neuron cell death in a mouse model of FALS is not mediated by the p53 cell survival regulator. AB - Mutant Cu/Zn superoxide dismutase (SOD1) associated with familial amyotrophic lateral sclerosis (FALS) causes selective motor neuron loss through unknown mechanisms of cell damage. Damaged neurons frequently undergo apoptosis mediated by the p53 cell survival regulator. We therefore studied whether motor neuron disease (MND) in mice expressing the human SOD1 mutant G93A is dependent on p53 by crossing G93A mice with p53-knockout mice. Since p53-/- mice's life expectance is usually shorter (160+/-49 days, n=11) than the time at which the G93A mice die from MND (212+/-50 days, n=7), only a few of the G93A/p53-/- double transgenics were expected to live to experience MND. Nevertheless, four of the 22 G93A/p53-/- mice succumbed to MND after 160+/-28 days, as expected under these conditions of competing death risks if the absence of p53 fails to protect from MND. Thus, MND in mice expressing G93A does not require p53. This conclusion is supported by histology: pre-symptomatic G93A mice display disease-associated vacuoles within the dendrites of motor neurons regardless of p53 status. PMID- 11011021 TI - The spinal 5-HT system contributes to the generation of fictive locomotion in lamprey. AB - Activation of NMDA receptors evokes sustained fictive locomotion in the isolated spinal cord of the sea lamprey Petromyzon marinus (P. marinus), but in the river lamprey Lampetra fluviatilis (L. fluviatilis) the ventral root activity is often irregular. A previous study showed that the number of 5-HT immunoreactive fibres, neurones and varicosities are much lower in the spinal cord of L. fluviatilis than in P. marinus. To further analyse the underlying mechanisms, the present study investigated the role of the 5-HT system in stabilising fictive locomotion. In P. marinus a blockade of 5-HT1A receptors by spiperone reversibly increased the frequency and the coefficient of variation. This implies that there is an endogenous release of 5-HT during fictive locomotion that is important for the generation of locomotor activity. In L. fluviatilis bath applied NMDA or D glutamate evoked in most cases irregular activity. An addition of 5-HT (0.5-2 microM) rapidly stabilised the burst generation and led to a sustained fictive locomotion. In a split-bath configuration, NMDA administered to the rostral part of the spinal cord in P. marinus evoked fictive locomotion in both the rostral part and the first few segments of the caudal part. When spiperone was added to the caudal part, the burst activity changed into tonic activity within 10 min. Taken together, these results indicate that activity in the intrinsic 5-HT system in the lamprey spinal locomotor network contributes significantly to the rhythm generation. The quantitative differences with regard to the 5-HT plexus between P. marinus and L. fluviatilis may account for the observed discrepancy between the two species. PMID- 11011022 TI - An in-vivo magnetic resonance imaging study of the olfactory bulbectomized rat model of depression. AB - The olfactory bulbectomized (OB) rat is a well-accepted animal model of depression. The present magnetic resonance imaging (MRI) investigation demonstrates alterations in signal intensities in cortical, hippocampal, caudate and amygdaloid regions in OB animals, but not in sham operated controls. Ventricular enlargement was also evident in OB animals. These alterations have implications with regard to the face and construct validity of this model. PMID- 11011023 TI - Retention of spatial information in hippocampally damaged rats overtrained on a cartographic task. AB - Hippocampal rats were overtrained on a cartographic task until they reached a performance equal to that of the control group. Twenty-four days later, during a retraining period, lesioned rats showed a profound retention deficit as compared to controls. However, Expt. 2 shows no retention deficit when a guidance strategy is used to acquire the spatial task. These results suggest that the hippocampus is crucial for long-term retention/consolidation of allocentric spatial information. PMID- 11011024 TI - Laterality, somatotopy and reproducibility of the basal ganglia and motor cortex during motor tasks. AB - We investigated the basal ganglia, motor cortex area 4, and supplementary motor area (SMA) using functional magnetic resonance imaging (fMRI) and five motor tasks: switching between finger and toe movements, writing, finger tapping, pronation/supination, and saccadic eye movements. We found reliable activation in the caudate nucleus and putamen in single subjects without the need for inter subject averaging. Percent signal changes in basal ganglia were smaller by a factor of three than those in SMA or motor cortex (1% vs. 2.5-3%). There was a definite foot-dorsal, hand-ventral basal ganglia somatotopy, similar to prior data from primates. Saccadic eye movements activated the caudate nucleus significantly more than the other tasks did. Unilateral movements produced bilateral activation in the striatum even when motor cortex activation was unilateral. Surprisingly, bilateral performance of the tasks led, on average, to consistently smaller basal ganglia activation than did unilateral performance (P<0.001), suggesting less inhibition of contralateral movements during bilateral tasks. Moreover, there was a striking dominance pattern in basal ganglia motor activation: the left basal ganglia were more active than the right for right handers, regardless of the hand used. This lateralization appears much stronger than that previously reported for motor cortex. Comparisons of inter-subject and intra-subject reproducibility indicated a much larger variability in basal ganglia and SMA compared to motor cortex, in spite of similar percent signal changes in the latter two structures. PMID- 11011025 TI - Intrathecal anti-IL-6 antibody and IgG attenuates peripheral nerve injury-induced mechanical allodynia in the rat: possible immune modulation in neuropathic pain. AB - Interleukin-6 (IL-6) is a pleiotrophic cytokine with a diverse range of actions including the modulation of the peripheral and central nervous system. We have previously shown significant IL-6 protein and messenger RNA elevation in rat spinal cord following peripheral nerve injury that results in pain behaviors suggestive of neuropathic pain. These spinal IL-6 levels correlated directly with the mechanical allodynia intensity following nerve injury. In the current study, we sought to determine whether it is possible to attenuate mechanical allodynia and/or alter spinal glial activation resulting from peripheral nerve injury by specific manipulation of IL-6 with neutralizing antibodies or by global immune modulation utilizing immunogamma-globulin (IgG). Effects of peripheral administration of normal goat IgG and intrathecal (i.t.) administration of IL-6 neutralizing antibody, normal goat or normal rat IgG on mechanical allodynia associated with L5 spinal nerve transection were compared. Spinal glial activation was assessed at day 10 post surgery by immunohistochemistry. Low dose (0.01-0.001 microg) goat anti-rat IL-6 i.t. administration (P=0.025) significantly decreased allodynia and trended towards significance at the higher dose (0.08 microg to 0.008 microg, P=0.062). Low doses (0.01-0.001 microg) i.t. normal goat and rat IgG significantly attenuated mechanical allodynia, but not at higher doses (0.08-0.008 microg; P=0.001 for both goat and rat IgG). Peripherally administered normal goat IgG (30 or 100 mg/kg) did not attenuate mechanical allodynia. Spinal glial activation was unaltered by any treatment. These data provide further evidence for the role of central IL-6 and neuroimmune modulation in the etiology of mechanical allodynia following peripheral nerve injury. PMID- 11011026 TI - Differential effects of stimulants on monoaminergic transporters: pharmacological consequences and implications for neurotoxicity. AB - Many psychostimulants alter plasmalemmal monoaminergic transporter function. Some, such as cocaine, prevent the reuptake of newly released dopamine, serotonin or norepinephrine into their associated neurons. Others, such as the amphetamines, facilitate release of these transmitters into the extraneuronal space by causing a reversal of function of these carrier proteins. An understanding of how psychostimulants regulate the function of not only plasmalemmal, but also vesicular monoamine transporter function is important to appreciate the pharmacological and sometimes neurotoxic consequences of administering these drugs, as well as the physiological regulation of these carrier proteins. Hence, this review will describe recent ex vivo studies investigating the rapid and differential affects of several stimulants on both plasmalemmal and vesicular monoamine transporter function. PMID- 11011027 TI - Nitric oxide donors and angiotensin-converting enzyme inhibitors act in concert to inhibit human angiotensin-converting enzyme activity and platelet aggregation in vitro. AB - This study investigates the effects of exogenous and endogenous nitric oxide (NO) on human circulating and endothelial angiotensin-converting enzyme activity and platelet aggregation. The NO donor S-nitroso-N-acetylpenicillamine (10(-8)-10(-6) M) significantly and dose-dependently inhibited serum angiotensin-converting enzyme activity. The concomitant addition of S-nitroso-N-acetylpenicillamine to angiotensin-converting enzyme inhibitor-treated (captopril or enalaprilat) serum, further reduced angiotensin-converting enzyme activity. In cultured endothelial cells from human umbilical veins (HUVECs), both S-nitroso-N-acetylpenicillamine and 3-morpholinosydnonimine (SIN-1) significantly reduced angiotensin-converting enzyme activity. An additative effect was seen with a combined treatment of captopril and S-nitroso-N-acetylpenicillamine. Treatment with the NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA) did not affect angiotensin converting enzyme activity. Thrombin inhibited endothelial angiotensin-converting enzyme activity, an effect that was abolished when cells were pretreated with L NMMA. Adenosine 5'-diphosphate (ADP)-induced platelet aggregation was inhibited with S-nitroso-N-acetylpenicillamine, SIN-1 and nitroglycerine. Captopril did not affect aggregation, while a high concentration of enalaprilat (10(-4) M) reduced it. The concomitant addition of 10(-5) M angiotensin-converting enzyme inhibitor to NO donor-treated platelets resulted in a further reduction of platelet aggregation. This effect was most evident with SIN-1 and enalaprilat. In conclusion, both exogenous and endogenous NO inhibit human angiotensin-converting enzyme activity. NO donors and angiotensin-converting enzyme inhibitors act in concert to inhibit angiotensin-converting enzyme and platelet aggregation. PMID- 11011028 TI - The chemotherapeutic oxaliplatin alters voltage-gated Na(+) channel kinetics on rat sensory neurons. AB - The chemotherapeutic oxaliplatin causes a sensory-motor neuropathy with predominantly hyperpathic symptoms. The mechanism underlying this hyperexcitability was investigated using rat sensory nerve preparations, dorsal root ganglia and hippocampal neurons. Oxaliplatin resulted in an increase of the amplitude and duration of compound action potentials. It lengthened the refractory period of peripheral nerves suggesting an interaction with voltage gated Na(+) channels. Application of oxaliplatin to dorsal root ganglion neurons resulted in an increase of the Na(+) current, a block of the maximal amplitude and a shift of the voltage-response relationship towards more negative membrane potentials. The effect was detectable on 13 of 18 tested cells. This observation, together with the absence of any effect on Na(+) currents of hippocampal neurons, suggests that the interaction of oxaliplatin is restricted to one or more channel subtypes. The effect of oxaliplatin could be antagonised by the Na(+) channel blocker carbamazepine which could be used to reduce side effects of oxaliplatin therapy in patients. PMID- 11011029 TI - BRL37344, but not CGP12177, stimulates fuel oxidation by soleus muscle in vitro. AB - The beta(3)-adrenoceptor agonist, (RR+SS)-(+/-)-4-[2-)2-)3-chlorophenyl)-2 hydroxyethyl)amino)propyl]ph enoxyacetate (BRL37344), stimulated fuel utilisation by isolated mouse soleus muscle at concentrations 10- to 100-fold lower than those required to stimulate lipolysis in brown adipocytes. At 1x10(-10) M BRL37344, uptake and phosphorylation of 2-deoxyglucose was increased (40%), as was glucose-oxidation (50%), palmitate-oxidation (70%) and oxidation of [2 14C]pyruvate (2-fold), indicating stimulation of tricarboxylic acid cycle reactions. Oxidation of [1-14C]pyruvate was unaffected, indicating no stimulation of pyruvate dehydrogenase activity. Other beta(3)-adrenoceptor agonists, disodium(RR)-5-[2-[[2-(3-chlorophenyl)-2-hydroxyethyl]-amino]propyl]- 1,3 benzodioxazole-2,2-dicarboxylate (CL316,243, 1x10(-7) M) and (S)-4-?2-[2-hydroxy 3-(4-hydroxyphenoxy)propylamino]ethyl?pheno xymeth ylcyclohexylphosphiric acid lithium salt (SB226552, 1x10(-9) M), achieved similar stimulation of 2 deoxyglucose uptake and phosphorylation but (+/-)-4-(3-t-butylamino-2 hydroxypropoxy)benzimidazol-2-one (CGP12177A) had no effect. The inhibitor of protein kinase A, H-89 (isoquinolinesulfonamide), had little effect on the stimulation of pyruvate-oxidation by BRL37344, while the specific inhibitor of protein kinase C, bisindolylmaleimide IX, reduced the stimulated rate to slightly below basal values. We consider that these responses provide evidence of the presence of a novel beta-adrenoceptor in skeletal muscle, which we have termed beta(skel)-adrenoceptor. PMID- 11011030 TI - A new structural class of subtype-selective inhibitor of cloned excitatory amino acid transporter, EAAT2. AB - We have studied the pharmacological effects of (RS)-2-amino-3-(3-hydroxy-5 methylisoxazol-4-yl)propionic acid (AMPA) and the enantiomers of (RS)-2-amino-3 (3-hydroxy-1,2, 5-thiadiazol-4-yl)propionic acid (TDPA) on cloned human excitatory amino acid transporter subtypes 1, 2 and 3 (EAAT1-3) expressed in Cos 7 cells. Whereas AMPA and (R)-TDPA were both inactive as inhibitors of [3H]-(R) aspartic acid uptake on all three EAAT subtypes, (S)-TDPA was shown to selectively inhibit uptake by EAAT2 with a potency equal to that of the endogenous ligand (S)-glutamic acid. (S)-TDPA thus represents a new structural class of EAAT2 inhibitor that will serve as a lead for the design of EAAT selective inhibitors. PMID- 11011031 TI - Effect of anti-inflammatory bowel disease drug, E3040, on urate transport in rat renal brush border membrane vesicles. AB - To confirm the assumption that 6-hydroxy-5, 7-dimethyl-2-methylamino-4-(3 pyridylmethyl)benzothiazole (E3040) acts on urate reabsorption by inhibiting urate-anion exchange at the luminal membrane of renal tubules, we investigated the inhibitory effect of E3040 and its two conjugated metabolites on hydroxyl ion (OH(-)) gradient-dependent urate uptake into brush border membrane vesicles from rat renal cortex and compared it with other uricosuric agents. The order of potency was AA193 (5-chloro-7, 8-dihydro-3-phenylfuro[2,3-g]-1,2-benzisoxazole-7 carboxylic acid)>benzbromarone>E3040>probenecid>E3040 sulfate>E3040 glucuronide. Furthermore, kinetic analysis revealed that E3040 may be a competitive inhibitor of the OH(-) gradient-dependent uptake of urate into brush border membrane vesicles. PMID- 11011032 TI - Inhibitors of tyrosine phosphatases block angiotensin II inhibition of Na(+) pump. AB - To determine how angiotensin II inhibits the Na(+) pump (Na(+), K(+)-ATPase) in rat zona glomerulosa, we selectively blocked signaling proteins that could be activated by the angiotensin AT(1) receptor and known to affect Na(+) pump activity. Inhibitors of protein kinase C [calphostin C (1 microM); staurosporine (1 microM)], phospholipase A(2) [arachidonyl triflouromethyl ketone (25 microM); quinacrine (75 microM)], diacylgycerol lipase [RHC-80267 (5 microM)], and tyrosine phosphorylation [tyrphostin 47 (100 microM)] had no effect on angiotensin II inhibition of the Na(+) pump. On the other hand, inhibitors of tyrosine phosphatases [phenylarsine oxide (5 microM) and 4-bromotetramisole oxalate (100 microM)] blocked angiotensin II inhibition, where as inhibitors of serine/threonine phosphatases [okadaic acid (1 microM) and microcystin (1.5 microM)] did not. Thus, angiotensin II inhibition of the Na(+) pump may in part be mediated by a tyrosine phosphatase. PMID- 11011033 TI - Effects of dopaminergic agents on carrageenan hyperalgesia in rats. AB - The present study explored the role of central dopaminergic transmission in a model of carrageenan-induced inflammatory pain by examining the effects of selective agonists and antagonists of dopamine receptors. The results were as follow: (1) LY171555 (trans-(-)-4aR-4,4a,5,6,7,8,8a,9-Octahydro-5-propyl-1H pyrazolo[3, 4-g]quinoline hydrochloride), dopamine D(2) receptor agonist, produced anti-hyperalgesia or hypoalgesia in the inflamed hindpaws and non inflamed hindpaws, respectively; spiperone hydrochloride (8-[4-(4-Fluorophenyl)-4 oxobutyl]-1-phenyl-1,3,8-triazaspiro[4, 5]decan-4-one hydrochloride), dopamine D(2) receptor antagonist, decreased the pain threshold of the non-inflamed hindpaws. (2) (+/-)-SKF38393 hydrochloride ((+/-)-1-Phenyl-2,3,4, 5-tetrahydro (1H)-3-benzazepine-7,8-diol hydrochloride), dopamine D(1) receptor agonist, produced anti-hyperalgesia or hypoalgesia when administered in a high dose (600 nmol), and decreased the pain threshold of non-inflamed hindpaws when administered in a low dose (150 nmol); R(+)-SCH23390 hydrochloride (R(+)-7-Chloro 8-hydroxy-3-methyl-1-phenyl-2,3,4, 5-tetrahydro-1H-3-benzazepine hydrochloride), dopamine D(1) receptor antagonist, induced anti-hyperalgesia or hypoalgesia, respectively. The present study suggests that the dopaminergic system is involved in the central modulation of inflammatory hyperalgesia, and that the different effects are probably induced by the different receptors. PMID- 11011034 TI - Effects of flurbiprofen and its enantiomers on the spinal c-Fos protein expression induced by noxious heat stimuli in the anaesthetized rat. AB - We have evaluated the effects of either intravenous or intraplantar administration of racemic-, S(+)- and R(-)-flurbiprofen on the spinal c-Fos protein expression after a single noxious heat stimulation (52 degrees C for 15 s) of the rat hindpaw in urethane anaesthetized rats. Two hours after noxious heat, numerous c-Fos protein immunoreactive (c-Fos-IR) nuclei (>70 c-Fos-IR nuclei per section at the level of L4-L5 segments) were observed with essential localization in the superficial (I-II) laminae of the spinal dorsal horn, i.e. areas containing numerous neurons driven exclusively by noxious stimuli. Considering the number of c-Fos-IR nuclei in laminae I-II, the intravenous injection of racemic-flurbiprofen (0.3, 3 and 9 mg/kg) was inefficacious and S(+) flurbiprofen had weak and non-dose-related effects. The same doses of R(-) flurbiprofen produced dose-related effects (r=0.58, P<0.05) with weak, but significant, effects for doses of 3 and 9 mg/kg (18+/-6% and 26+/-5% reduction of the number of noxious heat-evoked c-Fos-IR nuclei in laminae I-II, P<0.05 and P<0.01, respectively). The weak effects of R(-)-flurbiprofen are probably due to the central site of action since the intraplantar injection of a relatively high dose of 30 microg is inefficacious. These results provide further evidence for weak effects of non-steroidal anti-inflammatory drugs and their enantiomers on the acute responses to nociceptive stimulus which are very efficacious upon inflammatory nociception, but not upon brief noxious heat-evoked nociception. PMID- 11011035 TI - Role of endogenous endothelins in catecholamine secretion in the rat adrenal gland. AB - We investigated the role of endogenous endothelins in catecholamine secretion in response to transmural electrical stimulation in the retrogradely perfused rat adrenal gland. (R)2-[(R)-2-[(S)-2-[[1-(hexahydro-1H-azepinyl)]carbonyl]amino-4-++ +methy l-pentanoyl]amino-3-[3-(1-methyl-1H-indoyl)]propionyl]amino-3-(2-+ ++pyridyl) propionic acid (FR139317; 0.03-3 microM), an endothelin ET(A) receptor antagonist, inhibited the electrical stimulation-induced epinephrine and norepinephrine output. Neither N-cis-2, 6-dimethylpiperidinocarbonyl-L-gamma methylleucyl-D-1- methoxycarbonyl tryptophanyl-D-norleucine (BQ-788; 0.03-3 microM), an endothelin ET(B) receptor antagonist, nor phosphoramidon (1-100 mM), an endothelin-converting enzyme inhibitor, affected the catecholamine output responses. However, the inhibition by FR139317 of the catecholamine output responses was abolished by pretreatment with phosphoramidon (100 mM) or BQ-788 (3 microM). These results indicate that activation of endothelin ET(B) receptors by endogenous endothelins inhibits the catecholamine output responses under the condition in which endothelin ET(A) receptors are blocked. Exogenous endothelin-1 (1-100 nM) did not affect the catecholamine output responses, but it inhibited the responses under treatment with phosphoramidon and FR139317. Activation of endothelin ET(A) receptors may interfere with the endothelin ET(B) receptor mediated inhibitory action on the neuronally evoked secretion of adrenal catecholamines. PMID- 11011036 TI - The cyclo-oxygenase inhibitor nimesulide induces conditioned place preference in rats. AB - Two cyclo-oxygenase inhibitors, indomethacin and nimesulide, have been shown to potentiate morphine-induced stimulation of meso-accumbens dopamine neurons. In this study, an unbiased conditioned place preference procedure was used to evaluate whether nimesulide produces motivational effect after systemic administration in rats. These results show that nimesulide, at doses 0.1, 0.5 and 1 mg/kg, even lower than those usually applied for inflammatory conditions, induces conditioned place preference in rats, suggesting that it might possess rewarding properties in humans. PMID- 11011037 TI - Guanylate cyclase and not ATP-dependent K(+) channels seems temperature-dependent in smooth muscle relaxation of human umbilical arteries. AB - The effects of K(+) channel opener, nicorandil [N-(2-hydroxyethyl)-nicotinamide nitrate], on isolated human umbilical arteries were investigated at two temperatures: 37 degrees C and 25 degrees C. The purpose of this investigation was: (1) to confirm the relaxant effect of nicorandil, (2) to elucidate the influence of endothelium and temperature on nicorandil-induced relaxation, (3) to determine which of guanylate cyclase or ATP-sensitive K(+) channels was implicated in temperature-induced relaxation of smooth muscles. Rings, 3-mm-wide, were suspended in organ chambers for isometric force measurement. All solutions were aerated with 95% O(2)-5% CO(2) and maintained at 37 degrees C or 25 degrees C (cooling), pH 7.4. The presence of an intact endothelium was confirmed by immunohistochemistry. During the first set of experiments after contraction with 5-hydroxytryptamine (5-HT 10(-5) M), nicorandil (10(-9)-10(-4) M) was added to the organ chambers with controls and in with rings incubated with L-arginine, N nitro-L-arginine (L-NNA) an inhibitor of nitric oxide (NO) synthase, [1-H-(1,2,4) oxadiazole (4,3-a) quinoxalin-1-one] (ODQ), a specific inhibitor of guanylate cyclase, or glibenclamide, an antagonist of nicorandil, all at 10(-5) M. In another set of experiments, rings were contracted with 5-HT (10(-5) M) and relaxed with 3-morpholinosydnonimine [SIN-1 (10(-9)-3x10(-5) M) or cromakalim (10(-9)-3x10(-5) M)]. Our results showed that nicorandil induced concentration dependent relaxation. At 37 degrees C, in the control, the maximum relaxation was 90+/-5%, and 60+/-8% at 25 degrees C (P<0.01). However, the relaxation at 37 degrees C or 25 degrees C remained unchanged after pretreatment with L-arginine, L-NNA, this suggests that the same concentration of drugs used in this type of vessel does not appear to modulate the relaxant effect of nicorandil. On the other hand, we observed that the relaxant effect of SIN-1 was 72+/-5% at 37 degrees C and only 28+/-7% at 25 degrees C (P<0.01). However, relaxations with cromakalim were partly influenced by cooling. In the presence of ODQ, the nicorandil-induced relaxation observed at 37 degrees C or 25 degrees C was less than that in the control and in the rings incubated with glibenclamide. These results for human umbilical arteries indicate that cooling decreases the relaxation response of smooth muscles and that this is partly due to a decreased response to guanylate cyclase. PMID- 11011039 TI - Hemodynamic effects of MF 10058, a new cardioselective muscarinic M(2) receptor antagonist, in conscious dogs. AB - The 5-?4-[4-(diethylamino)butyl]-1-piperidinyl?acetyl-5H-dibenz[b, f]-azepine (MF 10058) is a new potent and selective muscarinic M(2) receptor antagonist. The hemodynamic effects of MF 10058 were investigated in conscious freely moving dogs. Placebo and three doses of MF 10058 (2, 4 and 8 mg/kg) were orally administered according to a randomised four-way crossover design. Heart rate, cardiac conduction times, systolic and diastolic blood pressure were telemetrically recorded for 12-24 h after dosing. After placebo administration, a consistent reduction over time in heart rate was observed during the night-time period (-15%, P=0.019). MF 10058 administration antagonised the nocturnal bradycardia and shortened QT interval. The effect of the drug reached statistically significance, compared to placebo, with the highest dose of 8 mg/kg (+19% on heart rate, P=0.013; -4% on QT interval, P=0.049). The effect on heart rate lasted for the entire 24-h observation period (+16%, P=0.030). Nocturnal systolic and diastolic blood pressure were not significantly affected by MF 10058. No other signs of peripheral or central cholinergic block were observed at any dose. The results of this study demonstrated that oral administration of MF 10058 produces long-lasting hemodynamic effects in the conscious dog. The drug has a therapeutic potential for the treatment of bradycardic disorders. PMID- 11011038 TI - Possible mechanism of cardioprotective effect of angiotensin preconditioning in isolated rat heart. AB - The present study is designed to investigate the mechanism of cardioprotective effect of angiotensin II preconditioning. Isolated perfused rat heart was subjected to global ischaemia for 30 min followed by reperfusion for 120 min. Coronary effluent was analysed for lactate dehydrogenase and creatine kinase enzyme release to assess the degree of cardiac injury. Myocardial infarct size was estimated macroscopically using triphenyltetrazolium chloride staining. Four episodes of angiotensin II preconditioning markedly reduced lactate dehydrogenase and creatine kinase release in the coronary effluent and decreased myocardial infarct size. Administration of prazosin (alpha(1)-adrenoceptor antagonist) before global ischaemia reduced the extent of ischaemia-reperfusion-induced myocardial injury. Moreover, administration of prazosin during angiotensin II preconditioning or depletion of biogenic amines by reserpinisation (0.5 mg/kg i.p.) did not affect the cardioprotective effect of angiotensin II preconditioning. On the other hand, colchicine (5 mg/kg i.p.) or polymyxin B (50 microM) treatment markedly attenuated the cardioprotective effect of angiotensin II preconditioning. On the basis of these results, it may be concluded that the cardioprotective effects of angiotensin II preconditioning may be mediated through protein kinase C and may not involve release of norepinephrine or activation of alpha(1)-adrenoceptor. PMID- 11011040 TI - Effect of a decrease in pH on responses mediated by P2 receptors in the rat mesenteric arterial bed. AB - The present study investigated the effect of acidosis (reduction in pH of the Krebs' solution from 7.4 to 6.9) on responses to vasoconstrictors and vasodilators, with a focus on purines, in the rat isolated perfused mesenteric arterial bed. alpha,beta-Methylene ATP (alpha,beta-meATP) (10 microM), a selective P2X receptor agonist, elicited a desensitizing vasocontraction, which was not significantly affected by a reduction in pH to 6.9. Contractions to ATP were also not significantly different at pH 6.9 compared to pH 7.4. In contrast, contractile responses to noradrenaline, methoxamine, and vasopressin were greatly attenuated at pH 6.9 (by 48-83%; P<0.01). At raised tone, vasorelaxations to ADP at P2Y receptors, and to calcitonin gene-related peptide (CGRP), were not different at pH 7.4 and pH 6.9. These data indicate that a reduction in pH (to 6.9) differentially affects responses to vasoconstrictors in the rat mesenteric arterial bed. There is no effect on contractions mediated via P2X receptors, but contractions to noradrenaline, methoxamine and vasopressin are greatly attenuated. PMID- 11011041 TI - Comparison of bisoprolol and carvedilol cardioprotection in a rabbit ischemia and reperfusion model. AB - Carvedilol, a selective alpha(1) and non-selective beta-adrenoceptor antagonist and antioxidant, has been shown to provide significant cardiac protection in animal models of myocardial ischemia. To further explore the mechanisms contributing to the efficacy of carvedilol cardioprotection, the effects of carvedilol on hemodynamic variables, infarct size and myeloperoxidase activity (an index of neutrophil accumulation) were compared with a beta(1) selective adrenoceptor antagonist, bisoprolol. Carvedilol (1 mg/kg) or bisoprolol (1 mg/kg) was given intravenously 5 min before reperfusion. In vehicle-treated rabbits, ischemia (45 min) and reperfusion (240 min) resulted in significant increases in left ventricular end diastolic pressure, large myocardial infarction (64.7+/-2.6% of area-at-risk) and a marked increase in myeloperoxidase activity (64+/-14 U/g protein in area-at-risk). Carvedilol treatment resulted in sustained reduction of the pressure-rate-index and significantly smaller infarcts (30+/-2.9, P<0.01 vs. vehicle) as well as decreased myeloperoxidase activity (26+/-11 U/g protein in area-at-risk, P<0.01 vs. vehicle). Administration of bisoprolol at 1 mg/kg resulted in a pressure-rate-index comparable to that of carvedilol and also decreased infarct size (48.4+/-2.5%, P<0.001 vs. vehicle, P<0.05 vs. carvedilol), although to a significantly lesser extent than that observed with carvedilol. Treatment with bisoprolol failed to reduce myeloperoxidase activity in the ischemic myocardial tissue. In addition, carvedilol, but not bisoprolol, markedly decreased cardiac membrane lipid peroxidation measured by thiobarbituric acid formation. Taken together, this study suggests that the superior cardioprotection of carvedilol over bisoprolol is possibly the result of carvedilol's antioxidant and anti-neutrophil effects, not its hemodynamic properties. PMID- 11011042 TI - Cytokine enhancement of endothelin ET(B) receptor-mediated contraction in human temporal artery. AB - Segments of human temporal artery were incubated in organ culture for 2 days in the absence or presence of cytokines. Thereafter, contractions were induced by the selective endothelin ET(B) receptor agonist sarafotoxin S6c, a peptide that does not induce contraction in fresh human temporal artery. Interleukin-1beta was most potent in increasing the sarafotoxin-induced contraction in cultured segments. Tumour necrosis factor (TNF)-alpha increased the magnitude of contraction to a similar degree, but at a higher dose. A significant increase was also induced by interferon-gamma, but not by interleukin-6 at the concentrations used. The results suggest that endothelin ET(B) receptor-mediated contraction can be enhanced by pro-inflammatory cytokines in a concentration-dependent manner, and this may have relevance for pathophysiological conditions where inflammation and vasoactivity are important. PMID- 11011043 TI - Myocardial contractility after infarction and carnitine palmitoyltransferase I inhibition in rats. AB - Inhibition of carnitine palmitoyltransferase I with etomoxir increases sarcoplasmic reticulum Ca(2+)-transport and V(1) isomyosin expression. To test whether etomoxir attenuates contractile dysfunction after myocardial infarction, we compared the contractility of papillary muscles from etomoxir- and placebo treated rats 6 weeks after infarction. Etomoxir induced cardiac hypertrophy in animals with small infarctions, and enhanced compensatory heart growth at large infarct size. Contractile function of papillary muscles from etomoxir-treated rats was improved particularly in animals with small infarctions. Thus, induction of mild cardiac hypertrophy by etomoxir in rats with small infarctions may be beneficial for myocardial performance. PMID- 11011044 TI - Tempol, a membrane-permeable radical scavenger, reduces dinitrobenzene sulfonic acid-induced colitis. AB - Inflammatory bowel disease is characterized by oxidative and nitrosative stress, leukocyte infiltration, and up-regulation of intercellular adhesion molecule 1 (ICAM-1) expression in the colon. The aim of the present study was to examine the effects of tempol, a membrane-permeable radical scavenger, in rats subjected to experimental colitis. Colitis was induced in rats by intracolonic instillation of dinitrobenzene sulfonic acid. Rats experienced bloody diarrhea and significant loss of body weight. At 4 days after the administration of dinitrobenzene sulfonic acid, the colon injury comprised of large areas of mucosal necrosis. Neutrophil infiltration (measured as increase in myeloperoxidase activity in the mucosa) was associated with up-regulation of ICAM-1 and expression of P-selectin and high levels of malondialdehyde (an indicator of lipid peroxidation). Immunohistochemistry for nitrotyrosine and poly (ADP-ribose) synthetase showed an intense staining in the inflamed colon. Treatment of rats with tempol (15 mg/kg daily i.p.) significantly reduced the appearance of diarrhea and the loss in body weight. This was associated with a remarkable amelioration of the disruption of the colonic architecture as well as a significant reduction in the degree of both neutrophil infiltration and lipid peroxidation in the inflamed colon. Tempol also reduced the appearance of nitrotyrosine and poly (ADP-ribose) synthetase immunoreactivity in the colon as well as the up-regulation of ICAM-1 and P selectin. The results of this study suggest that membrane-permeable radical scavengers, such as tempol, exert beneficial effects in experimental colitis and may, hence, be useful in the treatment of inflammatory bowel disease. PMID- 11011045 TI - Effect of a novel bifunctional endothelin receptor antagonist, IRL 3630A, on guinea pig respiratory mechanics. AB - This study characterized the in vitro pharmacological properties of a newly developed endothelin receptor antagonist, N-butanesulfonyl-[N-(3, 5 dimethylbenzoyl)-N-methyl-3-[4-(5-isoxazolyl)-phenyl]-(D)- alanyl]-( L) valineamide sodium salt (IRL 3630A), and its in vivo effects on respiratory mechanics were determined. IRL 3630A showed highly balanced affinities to human endothelin ET(A) and ET(B) receptors, giving apparent K(i) values of 1.5 and 1.2 nM, respectively. This compound also potently antagonized the endothelin-1 induced intracellular Ca(2+) increases in both embryonic bovine tracheal (EBTr) cells expressing endothelin ET(A) receptors and human Girardi heart (hGH) cells expressing endothelin ET(B) receptors. In guinea pig isolated tracheas having both endothelin ET(A) and ET(B) receptors, IRL 3630A greatly inhibited endothelin 1-induced contraction (pA(2)=7.1), which was partially or scarcely suppressed by the endothelin ET(A) receptor antagonist cyclo[-(D)-Trp-(D)-Asp-(L)-Pro-(D)-Val (L)-Leu-] (BQ-123) or the endothelin ET(B) receptor antagonist N-(3, 5 dimethylbenzoyl)-N-methyl-3-(4-phenyl)-(D)-phenylalanyl-(L)-t ryptop han (IRL 2500), respectively. Bolus i.v. injections of IRL 3630A administered into anaesthetized guinea pigs at 10 and 30 microg/kg inhibited endothelin-1 (1.3 microg/kg)-induced changes in respiratory resistance and compliance in a dose dependent manner, whereas both sodium 2-benzo[1, 3]dioxol-5-yl-4-(4-methoxy phenyl)-4-oxo-3-(3,4, 5-trimethoxy-benzyl)-but-2-enoate (an endothelin ET(A) receptor antagonist: PD 156707) and IRL 2500 at doses of up to 30 microg/kg did not affect endothelin-1-induced changes in respiratory mechanics, reflecting the in vitro results. IRL 3630A is thus an effective bifunctional endothelin receptor antagonist, and will be useful in clarifying the role of endothelin in pulmonary diseases such as bronchial asthma. PMID- 11011046 TI - Involvement of blockade of leukotriene B(4) action in anti-pruritic effects of emedastine in mice. AB - The anti-pruritic activity of emedastine difumarate was studied in mice. Emedastine (0.03-0.3 mg/kg) inhibited scratching induced by intradermal injection of histamine (100 nmol/site). Scratching induced by substance P (100 nmol/site) and leukotriene B(4) (0.03 nmol/site), but not by serotonin (100 nmol/site), was also suppressed by emedastine (0.03-0.3 mg/kg). Intradermal injection of substance P increased the cutaneous concentration of leukotriene B(4), which was not affected by emedastine. These results suggest that the inhibition by emedastine of substance P-induced itch-associated response is mediated by the blockade of leukotriene B(4) action. Anti-leukotriene B(4) action, as well as the anti-histamine action, may contribute to the anti-pruritic effects of emedastine. PMID- 11011047 TI - The 5-HT(2C/2B) receptor agonist, m-chlorophenylpiperazine, increases plasma glucagon levels in rats. AB - Effects of the 5-HT(2C/2B) receptor agonist m-chlorophenylpiperazine (mCPP) on plasma glucagon levels were investigated in rats. mCPP dose dependently increased plasma glucagon levels. Hyperglucagonemia elicited by mCPP was prevented by the 5 HT(2A/2B/2C) receptor antagonist, ritanserin, while the 5-HT(2A) receptor antagonist, ketanserin, did not show any effect. Increases in glucagon levels induced by mCPP were inhibited by prior adrenodemedullation. These results indicate that increases in plasma glucagon levels induced by mCPP are mediated by the 5-HT(2C/2B) receptor which in turn facilitates adrenaline release. PMID- 11011048 TI - Serum leptin levels after central and systemic injection of a serotonin precursor, 5-hydroxytryptophan, in mice. AB - Effects of peripheral and central injections of a serotonin (5-hydroxytryptamine, 5-HT) precursor, 5-hydroxytryptophan (5-HTP), on serum leptin levels were studied in mice. Intracerebroventricular (i.c.v.) injection of 5-HTP or 5-HT did not increase serum leptin levels, although the peripheral injection of 5-HTP elicited an apparent hyperleptinemia. The elevation of serum leptin levels in mice induced by the peripheral injection of 5-HTP was inhibited by pretreatment with the peripheral aromatic amino acid decarboxylase inhibitor, benserazide. Furthermore, the peripheral injection of 5-HT increased serum leptin levels. These results suggest that the hyperleptinemia following systemic injection of 5-HTP is elicited by 5-HT formed in the peripheral system. PMID- 11011050 TI - 2-Arachidonylglycerol, an endogenous cannabinoid, inhibits tumor necrosis factor alpha production in murine macrophages, and in mice. AB - 2-Arachidonylglycerol (2-AG) inhibits the production in vitro of tumor necrosis factor-alpha (TNF-alpha) by mouse macrophages, as well as in mice. It has no effect on the production of nitric oxide (NO). The effect on TNF-alpha is enhanced when 2-AG is administered together with 2-linoleylglycerol (2-Lino-G) and 2-palmitylglycerol (2-PalmG), an 'entourage effect' previously noted in several behavioral and binding assays. 2-AG also suppresses the formation of radical oxygen intermediates. PMID- 11011049 TI - Inverse agonism by Dmt-Tic analogues and HS 378, a naltrindole analogue. AB - The potent delta-opioid receptor antagonist H-2',6-L-tyrosine(Dmt)-1, 2,3,4 tetrahydroisoquinoline-3-carboxylic acid (Tic-OH) exhibited partial inverse agonism (EC(50)=6.35 nM, E(max)=-18.87%) for [35S]GTPgammaS binding and H-Dmt-Tic NH(2) was a neutral antagonist (no effect up to 30 microM). In contrast N,N(CH(3))(2)-Dmt-Tic-NH(2) was a full inverse agonist (EC(50)=2.66 nM, E(max)= 35.95%) similar to ICI 174864 ([N,N-diallyl-Tyr(1),Aib(2,3),Leu(5)]enkephaline) but with a 3.5-fold higher EC(50). In comparison, naltrindole was a neutral antagonist while its analogue HS 378 was a partial inverse agonist (E(max)= 12.99%). PMID- 11011051 TI - Transduction of the discriminative stimulus effects of zolpidem by GABA(A)/alpha1 receptors. AB - Zolpidem is an imidazopyridine with high affinity at gamma-aminobutyric acid(A) (GABA(A)) receptors expressing alpha1 subunits. In squirrel monkeys trained to discriminate a high dose of zolpidem (> or =3.0 mg/kg) from saline, zolpidem and another GABA(A)/alpha1 receptor-preferring agonist, zaleplon, substituted dose dependently for zolpidem, whereas the non-selective agonists diazepam and triazolam were did not substitute at any dose tested. These findings offer the first evidence for a selective role of GABA(A)/alpha1 receptors in the interoceptive effects of high doses of zolpidem. PMID- 11011062 TI - Neuromagnetic integrated methods tracking human brain mechanisms of sensorimotor areas 'plastic' reorganisation. AB - The potential for reorganization in the adult brain has been largely underestimated in the past and we are just beginning to understand the organisational principles involved in functional recovery. A bulk of experimental evidences have been accumulated in support of the hypothesis that neuronal aggregates adjacent to a lesion in the cortical brain areas can be progressively vicarious to the function of the damaged neurones. Such a reorganisation, if occurring in the affected hemisphere of a patient with a monohemispheric lesion, should significantly modify the interhemispheric symmetry of somatotopic organisation of the sensorimotor cortices, both in terms of absolute surfaces and number of "recruited" neurons, as well as of spatial coordinates. In fact, a roughly symmetrical organisation of sensorimotor - particularly for the hand contorl - in the right and left hemisphere has been observed in healthy humans by different methods of functional brain imaging, including fMRI, TMS, MEG, HD-EEG. Not uniform results about the functional brain activity related to sensory, motor and cognitive functions in normal and diseased subjects are often due to differences in the experimental paradigm designed as well as in the spatial and temporal resolution of the neuroimaging techniques used. The multi-modal integration of data obtained with several neuroimaging techniques allowed a coherent modelling of human brain higher functions. Functional magnetic resonance imaging (fMRI) provided fine spatial details (millimetres) of the brain responses, which were compared with the cortical maps of the motor output to different body districts obtained with transcranial magnetic stimulation (TMS). Magnetoencephalography (MEG) ability to study sensorimotor areas by analysing cortical magnetic fields, is also complementary to the motor cortex topographical mapping provided by TMS. MEG high temporal resolution allows to detect relatively restricted functional neuronal pools activated during cerebral processing of external stimuli. Moreover, these brain responses can be investigated with magnetoencephalography (MEG) and high density electroencephalography (EEG) techniques, with elevated time resolution (ms). With respect to the high resolution EEG technique, the MEG technique allowed a more precise localisation of the sites of neural activity buried into the cortical sulci, but was unable to detect the response of the crown of the cortical giri and of the frontal-mesial cortex (including the supplementary motor area), because of its poor sensitivity to radially oriented dipoles. The integration of functional and anatomical information provide cues on the relationship between brain activity and anatomic sites where this takes place, allowing the characterisation of the physiological activity of the cortical brain layers as well as to study the plastic reorganisation of the brain in different pathological conditions following stroke, limb amputation, spinal cord injury, hemisperectomy. PMID- 11011063 TI - Processing of multiple kinematic signals in the cerebellum and motor cortices. AB - The cerebellum and motor cortices are hypothesized to make fundamentally different but synergistic contributions to the control of movement. Richly interconnected, these structures must communicate and translate salient parameters of movement. This review examines the similarities and differences in the encoding of multiple limb movement parameters in the cerebellum and motor cortices. Also presented are recent data on direction and speed coding by cerebellar Purkinje cells and primary motor and dorsal premotor cortical neurons during a visually-instructed, manual tracking task. Both similarities and differences have been found in the way that these two motor areas process movement parameters. For example, the two motor control structures encode direction with almost identical depths of modulation, which may simplify the exchange of directional signals. Two major differences between the cerebellum and motor cortices consist of the distribution of the preferred directions and the manner in which direction and speed are jointly signaled within the discharge of individual neurons. First, an anterior-posterior distribution of preferred directions has been shown for both reaching and manual tracking, consistent with an intrinsic reference frame and/or the structure of afferent input. In contrast, neurons in the motor cortices have uniformly distributed preferred directions, consistent with general purpose directional calculations. Secondly, Purkinje cells in the cerebellum and motor cortices combine movement direction and speed information differently. For example, Purkinje cell discharge encodes combinations of direction and speed, a 'preferred velocity', while the motor cortical neurons use a temporal parcellation scheme to encode multiple parameters of movement. These results demonstrate that the cerebellum and motor cortices process and use kinematic information in fundamentally different ways that may underlie the functional uniqueness of the two motor control structures. PMID- 11011064 TI - Molecular mechanisms regulating the retrograde axonal transport of neurotrophins. AB - Neurotrophins are released from target tissues following neural innervation and bind to specific receptors situated on the nerve terminal plasma membrane. The neurotrophin-receptor complex undergoes retrograde axonal transport towards the cell soma, where it signals to the nucleus. This process allows neurotrophins to perform their numerous functions, which include the promotion of neuronal survival and the outgrowth of axons towards certain target tissues. The molecular events controlling each of the components of retrograde axonal transport are beginning to become defined. There is good evidence for the participation of phosphatidylinositol 3-kinase, phosphatidylinositol 4-kinase and the actin cytoskeleton in neurotrophin retrograde axonal transport in vivo. It also appears that the retrograde motor protein dynein mediates the retrograde axonal transport in vivo of neurotrophins such as nerve growth factor. This review discusses the role of the neurotrophin receptors in binding and axonal transport, the endocytic processes required for neurotrophin internalization, the targeting and trafficking of neurotrophins, and the propagation of neurotrophin-induced signals along the axon. PMID- 11011065 TI - Chemoarchitecture of the anuran auditory midbrain. AB - The anuran torus semicircularis consists of several subnuclei that are part of the ascending auditory pathway as well as audiomotor interface structures. Additionally, recent anatomical studies suggest that the midbrain tegmentum is an integral part of the audiomotor network. To describe the chemoarchitecture of these nuclei, taking into account the toral subdivisions, we investigated the distribution of serotonin, leucine-enkephalin, substance P, tyrosine-hydroxylase, dopamine D2-receptor, parvalbumin, aspartate, GABA, and estrogen-binding protein immunoreactivity in the midbrain of Bombina orientalis, Discoglossus pictus and Xenopus laevis. In the torus semicircularis, the highest density of immunoreactive fibers and terminals for all transmitters was found in the laminar nucleus. Parvalbumin-like immunoreactivity was highest in the principal nucleus, and D2-receptor-like immunoreactivity was uniformly distributed throughout the torus. In the tegmentum, axons and/or dendrites were stained with all antibodies except estrogen-binding protein. Additionally, heavily stained enkephalin and substance P-immunopositive fiber plexus were found in the lateral and dorsal tegmentum. The immunostainings revealed no qualitative differences between the three species. Immunopositive cell bodies were labeled in several brain areas, the connectivity of which with torus and tegmentum is discussed on the background of functional questions. The putative neuromodulatory innervation of both the laminar nucleus of the torus semicircularis and the tegmentum may be the anatomical basis for the influence of the animal's endogenous state on the behavioral reaction to sensory stimuli. These data corroborate earlier anatomical and physiological findings that the neurons of these nuclei are key elements in the audio-motor interface. PMID- 11011066 TI - Neurotrophic factors in Alzheimer's and Parkinson's disease brain. AB - The biomedical literature on the subject of neurotrophic growth factors has expanded prodigiously. This essay reviews neurotrophic factors (NTF) and their receptors in Alzheimer's disease (AD) and Parkinson's disease (PD) brain and recent updates on receptor signaling. The hypotheses for specific NTF involvement in neurodegenerative diseases in human and as potential therapy are based mainly on experimental animal and in vitro models. There are wide gaps in information on regional synthesis and cell contents of NTFs and their receptors in human brain. Observations on AD brain indicate increases in NGF and decreases in BDNF in surviving neurons of hippocampus and certain neocortical regions and decreases in TrkA in cortex and nucleus basalis. In PD brain, the few data available indicate decreases in neuronal content of GDNF and bFGF in surviving substantia nigra dopaminergic neurons. There are very few data regarding age-dependent effects on NTFs and on their receptors in human brain. Since NTFs in neurons are subject to retrograde and, in at least some cases, to anterograde transport from and to target neurons, their effects may be related to synthesis in local or remote sites or to changes in axoplasmic transport. Also, certain NTFs and their receptors are found to be expressed in activated glia. Thus, comparative in situ data for transcription levels and protein contents for NTFs and their receptors in both sites of neuronal origin and termination in human brain are needed to understand their potential roles in treating human diseases. PMID- 11011067 TI - Cortistatin: a member of the somatostatin neuropeptide family with distinct physiological functions. AB - Cortistatin is a recently discovered neuropeptide relative of somatostatin named after its predominantly cortical expression and ability to depress cortical activity. Cortistatin-14 shares 11 of the 14 amino acids of somatostatin-14 yet their nucleotide sequences and chromosomal localization clearly indicate they are products of separate genes. Now cloned from human, mouse and rat sources, cortistatin is known to bind all five cloned somatostatin receptors and share many pharmacological and functional properties with somatostatin including the depression of neuronal activity. However, cortistatin also has many properties distinct from somatostatin including induction of slow-wave sleep, apparently by antagonism of the excitatory effects of acetylcholine on the cortex, reduction of locomotor activity, and activation of cation selective currents not responsive to somatostatin. Expression of mRNA encoding cortistatin follows a circadian rhythm and is upregulated on deprivation of sleep, suggesting cortistatin is a sleep modulatory factor. This review summarizes recent advances in our understanding of the neurobiology of cortistatin, examines the similarities and differences between cortistatin and somatostatin, and asks the question: does cortistatin bind to a cortistatin-specific receptor? PMID- 11011068 TI - Female gonadal hormones, serotonin, and sexual receptivity. AB - The inhibitory and facilitatory effect of serotonergic (5-HT) receptor agonists and antagonists on the female rat lordosis reflex is reviewed. Emphasis is placed on the role of 5-HT(1A) and 5-HT(2) receptors. The effect of estrogen and progesterone on the lordosis response to 5-HT receptor-selective compounds is reviewed and potential mechanisms for hormonal modulation of the 5-HT system are suggested. Evidence that 5-HT modifies the female's position relative to a threshold for lordosis is presented. Finally, it is hypothesized that 5-HT's dual regulation of lordosis contributes to the female's ability to regulate mating behavior so that it occurs under physiological and environmental conditions that are conducive for individual, as well as species, survival. PMID- 11011069 TI - Adenosine: does it have a neuroprotective role after all? AB - A neuroprotective role for adenosine is commonly assumed. Recent studies revealed that adenosine may unexpectedly, under certain circumstances, have the opposite effects contributing to neuronal damage and death. The basis for this duality may be the activation of distinct subtypes of adenosine receptors, interactions between these receptors, differential actions on neuronal and glial cells, and various time frames of adenosinergic compounds administration. If these aspects are understood, adenosine should remain an interesting target for therapeutical neuroprotective approaches after all. PMID- 11011070 TI - The pharmacology of latent inhibition as an animal model of schizophrenia. AB - The nature of the primary symptoms of schizophrenia and our lack of knowledge of its underlying cause both contribute to the difficulty of generating convincing animal models of schizophrenia. A more recent approach to investigating the biological basis of schizophrenia has been to use information processing models of the disease to link psychotic phenomena to their neural basis. Schizophrenics are impaired in a number of experimental cognitive tasks that support this approach, including sensory gating tasks and models of selective attention such as latent inhibition (LI). LI refers to a process in which noncontingent presentation of a stimulus attenuates its ability to enter into subsequent associations, and it has received much attention because it is widely considered to relate to the cognitive abnormalities that characterise acute schizophrenia. Several claims have been made for LI having face and construct validity for schizophrenia. In this review of the pharmacological studies carried out with LI we examine its claim to predictive validity and the role of methodological considerations in drug effects. The data reviewed demonstrate that facilitation of low levels of LI is strongly related to demonstrated antipsychotic activity in man and all major antipsychotic drugs, both typical and atypical, have been shown to potentiate LI using a variety of protocols. Very few compounds without antipsychotic activity are active in this model. In contrast, disruption of LI occurs with a wide range of drugs and the relationship with psychotomimetic potential is less clear. Although reversal of disrupted LI has also been used as a model for antipsychotic acticity, mostly using amphetamine-induced disruption, insufficient studies have been carried out to evaluate its claim to predictive validity. However, like facilitation, it is sensitive to both typical and atypical antipsychotic agents. The data we have reviewed here demonstrate that facilitation of LI and, perhaps to a lesser extent, reversal of disrupted LI fulfil the criteria for predictive validity. PMID- 11011071 TI - Catecholamine systems in the brain of vertebrates: new perspectives through a comparative approach. AB - A comparative analysis of catecholaminergic systems in the brain and spinal cord of vertebrates forces to reconsider several aspects of the organization of catecholamine systems. Evidence has been provided for the existence of extensive, putatively catecholaminergic cell groups in the spinal cord, the pretectum, the habenular region, and cortical and subcortical telencephalic areas. Moreover, putatively dopamine- and noradrenaline-accumulating cells have been demonstrated in the hypothalamic periventricular organ of almost every non-mammalian vertebrate studied. In contrast with the classical idea that the evolution of catecholamine systems is marked by an increase in complexity going from anamniotes to amniotes, it is now evident that the brains of anamniotes contain catecholaminergic cell groups, of which the counterparts in amniotes have lost the capacity to produce catecholamines. Moreover, a segmental approach in studying the organization of catecholaminergic systems is advocated. Such an approach has recently led to the conclusion that the chemoarchitecture and connections of the basal ganglia of anamniote and amniote tetrapods are largely comparable. This review has also brought together data about the distribution of receptors and catecholaminergic fibers as well as data about developmental aspects. From these data it has become clear that there is a good match between catecholaminergic fibers and receptors, but, at many places, volume transmission seems to play an important role. Finally, although the available data are still limited, striking differences are observed in the spatiotemporal sequence of appearance of catecholaminergic cell groups, in particular those in the retina and olfactory bulb. PMID- 11011072 TI - Role of phospholipase A(2) on the variations of the choline signal intensity observed by 1H magnetic resonance spectroscopy in brain diseases. AB - Phospholipase A(2) catalyzes the hydrolysis of membrane glycerophospholipids leading to the production of metabolites observable by both 1H and 31P magnetic resonance spectroscopy. The signal of choline-containing compounds (Cho) observed by 1H magnetic resonance spectroscopy is constituted of metabolites of phosphatidylcholine, especially phosphocholine (PCho) and glycerophosphocholine (GPCho). The phosphomonoester (PME) and phosphodiester (PDE) signals observed by 31P magnetic resonance spectroscopy are, respectively, precursors and catabolites of phospholipids. A large number of brain diseases have been reported to cause variations in the intensity of the Cho, PME and PDE signals. Changes in the activity of phospholipase A(2) have been measured in many brain diseases. In this review, the relationships between the results of 1H and 31P magnetic resonance spectroscopy and the phospholipase A(2) assays are analyzed. In many brain diseases, the variation in the Cho signal intensity can be correlated with a stimulation or inhibition of the phospholipase A(2) activity. PMID- 11011073 TI - Local regulation of T cell numbers and lymphocyte-inhibiting activity in the interstitial tissue of the adult rat testis. AB - The testis is an immunologically privileged site, and transplantation data suggest that this privilege may be enhanced in cryptorchidism. Although alphabetaT cells, which mediate and promote the immune response, have access to the normal testis, relatively little is known about these cells in the abdominally located testis. An increase in testicular lymphocyte-inhibiting cytokines has also been implicated in enhanced graft survival following the experimental induction of cryptorchidism. Consequently, T cell traffic and lymphocyte-inhibiting activity in testes of cryptorchid adult rats were examined in the following study. Numbers of alphabetaT cells and the cytotoxic CD8(+) T cell subset in the testis were unaffected following 1 month of cryptorchidism. In contrast, subcutaneous testosterone implants, which inhibit Leydig cell function through suppression of gonadotrophin secretion, reduced these parameters in both scrotal and abdominal testes. Testicular T cell numbers were positively correlated with the number of testicular resident macrophages, which also were reduced by subcutaneous testosterone implants. The concentration of lymphocyte inhibiting activity in the testicular interstitial fluid was reduced by 80% in short-term (1 month) and longer-term (3 months) cryptorchidism. These data indicate that the T cell population, and in particular the CD8(+) T cell subset, in the rat testis is functionally related to the resident macrophages or Leydig cells. On the other hand, testicular lymphocyte-inhibiting activity does not appear to be a determinant of the number of testicular T cells, and may not be a major factor in the prolonged survival of certain grafts in the abdominal testis. PMID- 11011074 TI - Ability of recombinant human TNF binding protein-1 (r-hTBP-1) to inhibit the development of experimentally-induced endometriosis in rats. AB - The aim of this study was to assess whether r-hTBP-1 (recombinant human tumor necrosis factor binding protein-1), the soluble form of tumor necrosis factor alpha (TNF) receptor type 1 might be effective in counteracting the proliferation of ectopic endometrium using an in vivo experimental model of endometriosis. The in vivo model involved transplanting a square fragment of autologus uterine tissue onto the inner surface of the abdominal wall in rats. r-hTBP-1 was administered for 1 week at 10 mg/kg, s.c. divided into two daily injections. The gonadotropin-releasing hormone antagonist antide was used for reference and given at the dose of 2 mg/kg, s.c. every 3 days for 1 week. The animals were killed 2 and 9 days after the last treatment and the size of endometriotic implants measured. Blood samples and spleens were also taken for assessment of estradiol 17beta levels and natural killer (NK) activity in vitro against murine YAC cells, respectively. The results of this study indicate that r-hTBP-1 is effective in reducing the size of the endometriotic-like foci mainly at the later sacrifice time-point when they were significantly decreased by 64% as compared to control animals. As expected, antide induces an almost complete and statistically significant remission both at the 2-day (94%) and the 9-day (88%); sacrifice time point. Histological examination indicates that, compared to controls, r-hTBP-1 induces a slightly increased degeneration of the stromal tissues of the implants at both examination times and, limitedly to the earlier observation time, of the mucosal epithelium. No differences in the spleen cell NK activity were observed at either sacrifice time-points in any treatment group. Estradiol-17beta concentrations are significantly decreased in the antide-treated groups only at 9 days while no statistically significant changes are found in the animals receiving r-hTBP-1. The results of this study carried out in a rat experimental model of endometriosis provide evidence of the potential effectiveness of r-hTBP 1 in this pathological condition and support the role of TNF in its development. PMID- 11011075 TI - Activation and deposition of human breast-milk complement C3 opsonins on serum sensitive Escherichia coli 0111. AB - Little is known about the physiological roles and contribution of the human breast-milk complement system in the protection of both the maternal mammary gland and the nursing infant. The ability of a serum-sensitive Escherichia coli to activate the complement components of human breast-milk and colostrum was assessed in vitro. The consequent deposition of C3 fragments, C3b, iC3b and C3dg, on the bacteria was analysed, using a slight modification of a standard ELISA technique for the assessment of activated C3 fragments. The deposition C3 fragments from human milk were demonstrated on the killed bacteria, E. coli NCTC 8007, serotype 0111 K58(B4) H2, using buffers with and without detergent, supporting both the classical and alternative pathways of complement activation. The milk fat appears to competitively inhibit the deposition of these opsonins on the solid-phase bacteria. This study suggests that the complement system is able to contribute to the increased resistance of breast-fed infants against infections. PMID- 11011076 TI - Simplified procedure for detection of enteric pathogenic viruses in shellfish by RT-PCR. AB - Epidemiological evidence linking the transmission of enteric viral disease to shellfish has been known for a long time. A variety of methods have been described for the detection of viral contaminants in shellfish using RT-PCR. However, these methods generally include numerous, often fastidious and time consuming steps for virus release from shellfish tissues and viral RNA isolation. A simplified procedure based on the enzymatic liquefaction of shellfish digestive tissues without any mechanical homogenisation step, followed by a simple clarification of the lysate using dichloromethane extraction, was developed. Viral RNA is isolated directly from the shellfish extract by a guanidium thiocyanate-silica extraction method, adapted for the use of a vacuum manifold system. Virus-specific RT-PCR assays were set up for detection of genomic sequences of the predominant viral pathogens, HAV, Astrovirus and Norwalk-like viruses (from genogoups I or II). The specificity of the amplicons is confirmed finally by hybridisation with DIG-labelled specific probes. The overall procedure applied to shellfish samples spiked with HAV particles allowed a detection of 20 pfu of HAV per g of hepatopancreas. In addition, up to 20 samples can be tested within 24 h. PMID- 11011077 TI - HCV RNA levels in hepatocellular carcinomas and adjacent non-tumorous livers. AB - To determine the antiviral effects of drugs targeted to hepatitis C virus (HCV) in chronic hepatitis patients, an accurate quantitative method with high sensitivity is needed. Reverse transcription nested polymerase chain reaction (RT PCR) is the most sensitive method for the detection of HCV sequences in clinical specimens. However, this method is not quantitative. For this purpose, a quantitative competitive assay was developed that combines RT and PCR followed by image analysis to quantify HCV RNA. This assay targets the highly conserved 5' non-coding region of HCV and is based on the co-amplification of wild type HCV RNA with known amounts of mutant synthetic RNA. The mutant internal control used in these experiments differs from the wild type RNA by two nucleotide substitutions, which introduces an internal restriction enzyme site. In this report, this method was used to determine the levels of positive strand RNA in 11 HCV positive hepatocellular carcinomas (HCC) and compared these with adjacent non tumorous liver tissue. To confirm that the difference in viral titers is not related to variations in the amount of RNA used in the assay, glyceraldehyde 3 phosphate dehydrogenase (GAPDH) mRNA was also assessed by competitive RT-PCR in all tissue extracts. Using this competitive assay it was determined that HCV RNA levels in the liver and tumor samples ranged from 10(3) to 10(6) molecules per microg of total RNA which is similar to previous reports. Interestingly, the amount of HCV in all the non-tumorous liver specimens were found to be significantly higher (P<0.05) than the surrounding tumors, while the GAPDH mRNA levels were found to be similar in both liver and tumor. Competitive RT-PCR is a sensitive, accurate and reliable method to determine HCV titers in clinical specimens. Using this method it was determined that malignant tumor cells harbor less HCV as compared with the surrounding non-tumorous liver cells. PMID- 11011078 TI - The use of phylogenetic data to develop molecular tools for the detection and genotyping of Yam mosaic virus. Potential application in molecular epidemiology. AB - Molecular detection of Yam mosaic virus (YMV), the most important potyvirus for yam plants (Dioscorea spp.) has been limited by its high genetic diversity. According to recent phylogenetic data, suitable molecular tools were developed for detection and genotyping. A reliable and rapid molecular test is described using single-tube immunocapture (IC)-RT-PCR combining the use of a monoclonal antibody able to recognise all YMV serotypes and primers designed to take into account the whole variability of YMV. Phylogenetic data were then used to develop an uncoupled IC-RT-PCR procedure adapted to the genotyping of the three major phylogenetic groups and the two recombinants implicated in the YMV epidemics in the Caribbean and French Guiana. A third exploitation of phylogenetic data was carried out with the development of a typing test based on the analysis of short nucleotide sequences. The direct sequencing of a 450 nts fragment constituted a reliable complementary typing tool as the resulting information was quite similar to that obtained with sequences of the complete coat protein gene. The approach, consisting in a robust study of YMV variability followed by the use of phylogenetic data to develop reliable detection and genotyping tools, offers new perspectives for powerful molecular epidemiological studies. PMID- 11011079 TI - Improved RNA extraction and one-tube RT-PCR assay for simultaneous detection of control plant RNA plus several viruses in plant extracts. AB - A procedure was developed for simultaneous detection of plant RNA viruses and of plant RNA, as a control. RT-PCR amplification with primers designed for the detection of the plant mRNAs encoding malate dehydrogenase (MDH) and the large subunit of ribulose bisphosphate carboxylase oxygenase (RubiscoL) was used for the development of a plant extraction procedure that consistently yields extracts that can be amplified. The control amplification was used successfully on extracts from cane, leaf and/or bud tissues from grapevine, apple, raspberry, strawberry, peach, apricot, plum and wheat. Multiplex RT-PCR conditions were established for the simultaneous detection in grapevine extracts of either arabis mosaic virus, rupestris stem pitting associated virus and malate dehydrogenase mRNA, or grapevine virus A, grapevine virus B, grapevine leafroll associated virus-3, and RubiscoL mRNA. PMID- 11011080 TI - Determination of tumor necrosis factor-alpha levels in dengue virus infected patients by sensitive biotin-streptavidin enzyme-linked immunosorbent assay. AB - A modified sandwich enzyme-linked immunosorbent assay using biotin-streptavidin system (BS-ELISA) was developed to determine levels of tumor necrosis factor alpha (TNF-alpha) in serum samples of children infected with dengue virus (n=99) and healthy controls (n=41). The minimum detectable concentration of TNF-alpha by the BS-ELISA was 3.3 pg/ml. The mean TNF-alpha level was highest in those patients with dengue shock syndrome (DSS) or dengue hemorrhagic fever (DHF) grade III (37.44+/-42.0 pg/ml). Lower levels were found in DHF grade I (28.44+/-42.7 pg/ml), DHF grade II (24. 21+/-25.4 pg/ml) and dengue fever (DF) (14.10+/-24.0 pg/ml). TNF-alpha in the sera of DF and DHF patients could be detected on days 2 6 after the onset of fever, the high level occurring on day 5. TNF-alpha was detected in 41.4% (24.01+/-35.2 pg/ml) of dengue virus infected patients and 7.3% (4.2+/-15.6 pg/ml) of control subjects. The sera of patients contained significantly higher levels of TNF-alpha than the sera of controls, P value<0.001. DHF patients had significantly higher levels of TNF-alpha than DF patients (P-value=0.020) but no difference in the TNF-alpha levels from sera of DHF grades I-III patients was observed (P-value=0.295). The results indicate that the BS-ELISA is a very sensitive method for determining TNF-alpha in serum samples of DF and DHF patients. The TNF-alpha levels might be associated with dengue virus infection and related to disease severity of DHF. PMID- 11011081 TI - A comparative analysis of freon substitutes in the purification of reovirus and calicivirus. AB - Freon 113 (Freon) is an essential component used in some viral purification methods to separate virus from infected cell debris. With its environmental and toxic hazards, Freon's availability is limited and more tightly regulated. Several organic solvent substitutes were selected to identify a suitable Freon replacement for the purification of both cultivable reovirus and fastidious calicivirus. Reovirus was extracted from tissue cultured cells with each solvent tested and purified in cesium chloride gradients by standard techniques. Purified virions were analyzed for conservation of physical and biological properties by morphological examination and infectivity studies. The purification of calicivirus nucleic acid from stool samples using selected solvents was also examined. Solvent-extracted calicivirus RNA was reverse transcribed and quantified by polymerase chain reaction amplification of a standard diagnostic 117 bp amplicon. These studies indicated that Vertrel XF (a newly developed environmentally friendly Freon substitute) and a 7:3 mixture of isopentane/1 chlorobutane are suitable replacements. Considerations of flammability and ease of use suggest that Vertrel XF is the preferred choice as a Freon substitute for the purification of these non-enveloped viruses. PMID- 11011082 TI - Development of an immunomagnetic capture reverse transcription-PCR assay for the detection of Norwalk virus. AB - Norwalk virus (NV) is the prototype human virus of the family Caliciviridae. A rapid immunomagnetic capture/reverse transcription-(IMC/RT-)PCR assay was developed for the detection of NV. Immunomagnetic capture (IMC) utilizes paramagnetic beads coupled to a virus-specific antibody and allows separation of virus from contaminating materials and virus concentration in a single step. The detection limit of the developed assay was approximately 250-750 genomic equivalents/ml of 10% stool suspension. The detection limit of the assay was not altered by the presence of excess hepatitis A virus (HAV), although non-specific binding of HAV to the paramagnetic beads was observed. A panel of 100 stools from experimental human infections was screened for NV using a previously described heat release method, an antigen ELISA, or IMC/RT-PCR. NV was detected in 65/100 of these samples by IMC/RT-PCR compared to only 38/99 by heat release and 31/95 by antigen detection ELISA. All samples that were negative by IMC were also negative by both heat release and antigen ELISA. The number of samples in which RT-PCR was inhibited was greatly reduced by the use of IMC/RT-PCR compared to the heat release method (1/100 and 16/95 samples inhibited, respectively). The ability of IMC to concentrate virus (> or =2000-fold greater than heat release) and effectively remove inhibitory substances gives this assay distinct advantages over both the heat release and antigen ELISAs. PMID- 11011083 TI - Development of an RT-PCR diagnostic test for an avian astrovirus. AB - Astroviruses are small round viruses that cause enteric disease in the young of several species. Detection and diagnosis of astrovirus infection in non-human hosts relies heavily on electron microscopy and fluorescent antibody tests. Recently, our laboratory isolated and sequenced an avian astrovirus from poult enteritis mortality syndrome affected turkeys. These studies describe the development of RT-PCR methods, which specifically detect regions of the viral capsid and polymerase genes, and demonstrate their use in detecting astrovirus infection in commercial turkey flocks. PMID- 11011084 TI - Packaging of small molecules into VP1-virus-like particles of the human polyomavirus JC virus. AB - Recombinantly expressed VP1-virus-like particles (VP1-VLP) of human polyomavirus JC virus (JCV) were described recently as a new DNA transporter system. It was shown that DNA molecules could be packaged into VP1-VLP during a controlled chemical reassociation/dissociation process. In the present study VP1-VLP were studied as carriers for pharmaceutical substances. Propidium iodide (PI) was packaged into VP1-VLP as a reporter molecule. The PI-containing VP1-VLP could be detected directly by flow cytometry. The fluorescence intensity of the VP1-VLP depended strongly on the initial PI concentration. This packaging method is easy to handle and applicable to viruses and VP1-VLP which can be dissociated and reassociated chemically. PMID- 11011085 TI - Use of electron microscopic and immunogold labeling techniques to determine polyomavirus recombinant VP1 capsid-like particles entry into mouse 3T6 cell nucleus. AB - Murine polyomavirus major structural protein VP1 could assemble into capsid-like particles when expressed in the baculovirus system. The recombinant capsid-like particles that were purified by CsCl density gradient centrifugation were capable of packaging host DNA. Electron microscopic and immunogold labeling techniques were used to study the entry of these VP1 recombinant capsid-like particles into mouse 3T6 cells. It was found that these VP1 recombinant capsid-like particles, which lack polyomavirus minor structural proteins (VP2 and VP3), use the same mechanism to enter mouse 3T6 cell cytoplasm and nucleus as that used by native polyomavirus virions. PMID- 11011086 TI - Elimination of HIV-1 plasmid DNA from virus samples obtained from transfection by calcium-phosphate co-precipitation. AB - Molecular genetics is a powerful tool to analyze the replication cycle of human immunodeficiency virus type 1 (HIV-1). Culture fluids obtained from HIV-1 plasmid transfected cells by calcium-phosphate co-precipitation were treated with ethyleneglycol bis (beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and DNase I to obtain HIV-1 stocks virtually free of input plasmid DNAs. Even after amplification by polymerase chain reaction (PCR), no plasmid DNA was detected in cells following infection with EGTA/DNase I-treated virus samples. This method is particularly useful for the examination of the early replication phase of HIV-1 by PCR. PMID- 11011087 TI - Leaf secretion composition of the mangrove species Avicennia germinans (L.) in relation to salinity: a case study by using total-reflection X-ray fluorescence analysis. AB - The aim here was to use total-reflection X-ray fluorescence (TXRF) to determine the elemental composition (besides Na), and the relative contribution of each element in leaf secretion of the mangrove species Avicennia germinans, grown under contrasting salinities (0-865 mol m(-3) NaCl). Na was determined by conventional atomic absorption spectrometry (AAS). Total secretion was 2.00+/ 0.28 mmol m(-2) per day in control plants (at 0 NaCl) and increased progressively up to 46.87+/-7.14 mmol m(-2) per day at 865 mol m(-3) NaCl. Na and Cl accounted for 85.91+/-1.81% of the total secretion in control plants and about 96.32+/ 0.30% in salt-treated plants. The excretion of Na exceeded that of Cl when salinity increased and this led to a progressive increase in Na/Cl ratios from 0.46+/-0.02 in control plants up to 2.75+/-0.42 (at 865 mol m(-3) NaCl). Other elements were also secreted in sizeable amounts such as K, S, Ca, Br and Zn. However, the relative importance of these elements, in terms of total secretion, was considerable in control plants (15% of the total secretion) and declined significantly under salinity treatments (<5% of the total secretion). In conclusion, TXRF has been shown to be a powerful tool allowing quantitative determination of Cl (1.2-13 mmol m(-2) per day), secreted in relatively large quantities, as well as other elements secreted in intermediate (S, Ca and K; 0.07 1.00 mmol m(-2) per day) and in trace quantities (Br and Zn; 0.6-4 umol m(-2) per day). PMID- 11011088 TI - An efficient method for the production of transgenic plants of peanut (Arachis hypogaea L.) through Agrobacterium tumefaciens-mediated genetic transformation. AB - Cotyledon explants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (>90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and beta-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of transgenic plants. Transformed individuals were obtained through selection on medium containing 125 mg l(-1) kanamycin. A large number of independently transformed plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of transgenic plants. Analysis of 35 transgenic plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120-150 days were required between the initiation of explant transformation and transfer of rooted plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation. PMID- 11011089 TI - In vitro plant regeneration in Macroptilium atropurpureum, a legume with a broad symbiont range for nodulation. AB - Macroptilium atropurpureum is a model legume with a broad symbiont range for nodulation. We have achieved the first in vitro plant regeneration of this species using cv. Siratro. Hypocotyl explants excised from dark-grown seedlings generated slimy, friable calli after three weeks' culture on B5 medium containing 1-2 mg/l kinetin and 0.05 mg/l alpha-naphthaleneacetic acid. This was followed by the generation of green organogenic callus with shoot buds by subculturing the explants to hormone-free B5 medium 20 days after the start of culture. The green organogenic calli with shoot buds were maintained as organogenic callus by subculturing on the same medium, and shoots were elongated on hormone-free B5 medium. Elongated shoots were rooted on half-strength B5 medium. Most regenerated plants were morphologically normal, diploid and fertile, although tetraploid plants appeared at a low frequency (8%). PMID- 11011090 TI - Partial characterization and localization of a novel type of antifungal protein (IWF6) isolated from sugar beet leaves. AB - An antifungal protein was isolated from the intercellular washing fluid (IWF) of leaves of sugar beet (Beta vulgaris L., cv. Monova) and purified to homogeneity. The protein, IWF6, comprising 37 amino acids with six cysteines, was able to inhibit the growth of the pathogen Cercospora beticola (Sacc.) in vitro, by 75% after 120 h of growth at a concentration of 20 ug ml(-1). The amino acid sequence data were used to generate a polymerase chain reaction (PCR) clone, employed for the isolation of a corresponding cDNA clone. The cDNA encodes a precursor protein with an N-terminal putative signal sequence of 45 amino acids, followed by the mature protein of 37 amino acids. Antibodies raised against a synthetic peptide covering the complete sequence of IWF6 were used in immunolocalization studies. The protein was recognized by the antibody in nearly all leaf cell types except epidermal cells. In necrotic tissue, the protein was mainly recognized on C. beticola hyphae growing in a 'pellet' (ball-like) structure. The hyphal 'pellets' are primarily located beneath the stomata. IWF6 shows less than 26% identity to any previously described protein. PMID- 11011091 TI - Pepper gene encoding a basic class II chitinase is inducible by pathogen and ethephon. AB - A chitinase cDNA clone (designated CAChi2) was isolated from the cDNA library of pepper leaves infected with Xanthomonas campestris pv. vesicatoria. The 1004-bp full-length CAChi2 cDNA encodes a basic chitinase with an N-terminal 24 amino acid signal peptide followed by a catalytic region. An analysis of its sequence indicates that CAChi2 is a class II chitinase, because it does not have chitin binding domain and C-terminal extension sequences. The deduced amino acid sequence of CAChi2 has a high level of identity with class II chitinases from potato, tomato, tobacco and petunia. Southern analysis demonstrated that the CAChi2 chitinase is encoded by a single or two copy genes in the pepper genome. Following X. campestris pv. vesicatoria or Phytophthora capsici infection, the CAChi2 chitinase mRNA was more highly expressed in the incompatible interaction, compared to expression in the compatible interaction. Treatment with ethylene releasing ethephon resulted in a strong accumulation of the transcripts in the leaves. In contrast, DL-beta-amino-n-butyric acid, salicylic acid and methyl jasmonate were not effective in inducing CAChi2 transcripts in pepper leaves. PMID- 11011092 TI - Phytochrome and post-translational regulation of nitrate reductase in higher plants. AB - The possible influence of phytochrome on the activity state of nitrate reductase (NR) was investigated in etiolated plants where the expression of the NR gene is known to be under the control of phytochrome. Activity state is defined as NR activity assayed in the presence of Mg(2+) as percentage of NR activity measured in the absence of Mg(2+). This measurement is assumed to reflect non phosphorylated NR as percentage of total NR. Beside etiolated barley and maize leaves, a photosynthetic mutant of Lemna aequinoctialis was investigated and compared with the wild type. The increase of NR activity following a red light pulse, mediated via phytochrome, was confirmed in all etiolated plant species investigated as well as in both strains of L. aequinoctialis cultivated in glucose-containing medium. The effect of continuous red light surpassed the effect of a single red light pulse in each case. However, the results did not show any stimulating effect of phytochrome on the activity state caused by post translational modulation. The activity state was strongly increased by continuous red light in the wild type of L. aequinoctialis but not in the photosynthetic mutant. These results show that the phytochrome system is not important for the post-translational regulation of NR. PMID- 11011093 TI - Growth promoting effect of two Sinorhizobium meliloti strains (a wild type and its genetically modified derivative) on a non-legume plant species in specific interaction with two arbuscular mycorrhizal fungi. AB - In the present study, we have investigated whether the ubiquitous rhizosphere soil organism Sinorhizobium meliloti has a plant growth promoting (PGP) effect on non-leguminous plant species. Such PGP activity was investigated for both a wild type strain and its genetically modified (GM) derivative, which had an enhanced biofertilizer capability. The PGP effect of these rhizobial strains was tested in interaction with two arbuscular-mycorrhizal (AM) fungi: G. mosseae or G. intraradices on lettuce (Lactuca sativa L.) plants. Both rhizobial strains were efficient in increasing lettuce biomass and also induced modifications on root morphology, particularly in mycorrhizal plants; thus these strains behave as plant growth promoting rhizobacteria. In non-mycorrhizal plants, both strains exhibited a similar growth promoting effect on lettuce. However, both rhizobial strains differed in mycorrhizal plants with regard to (i) biomass production, (ii) the length of axis and lateral roots, and (iii) the number of lateral roots formed; effects which were, in turn, affected by the AM fungus involved. Microbial treatments were more effective on root growth and morphology at earlier developmental stages (20 days of plant growth) but, in a later stage (after 40 days), the microbial effects were more relevant at increasing plant biomass. The interaction between the GM rhizobial strain and G. mosseae produced the highest growth promoting effect (476% over control), in spite of the fact that G. intraradices showed a quicker and higher colonization ability than G. mosseae. Microbial interactions inducing PGP effects did not benefit AM colonization nor the succinate dehydrogenase activity in the AM fungal mycelium. Irrespective of the underlying mechanisms, which are being now investigated, the interactions between rhizobial strains, as free-living saprophs, and AM fungi are noteworthy, and depend on the microbial genotype involved. PMID- 11011094 TI - A DNA-binding activity for the promoter of the gene encoding C(4) phosphoenolpyruvate carboxylase is modulated by phosphorylation during greening of the Sorghum leaf. AB - Electrophoresis mobility shift assay (EMSA) identified nuclear proteins with binding activity to a 430 bp promoter fragment of the Sorghum C(4) phosphoenolpyruvate carboxylase gene (SvC4). The DNA binding activities (two main retarded bands; PC1 and PC2) were high in nuclear extracts from etiolated leaves, decreased during greening and became very low or null in nuclear extracts from green leaves. This process was found to be mediated by phytochrome and was apparently irreversible since the DNA-binding activities were not restored in green plants kept in continuous darkness. The AT-rich region of the promoter fragment was identified to be the interaction domain of PC2. The detection of PC2 with EMSA was markedly reduced by preincubation of nuclear protein extracts with Mg-ATP or Mg-GTP and restored in the presence of a general protein serine/threonine-kinase inhibitor, K252a. The results suggested that the PC2 binding activity was modulated by phosphorylation during the greening process of the Sorghum leaf. PMID- 11011095 TI - Chilling stress-induced changes of antioxidant enzymes in the leaves of cucumber: in gel enzyme activity assays. AB - To investigate the antioxidant defense system, chilling stress-induced changes of antioxidant enzymes were examined in the leaves of cucumber (Cucumis sativus L.). Chilling stress preferentially enhanced the activities of the superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and peroxidase specific to guaiacol, whereas it induced the decrease of catalase activity. In order to analyze the changes of antioxidant enzyme isoforms against chilling stress, foliar extracts were subjected to native PAGE. Leaves of cucumber had four isoforms of Mn-SOD and two isoforms of Cu/Zn-SOD. Fe-SOD isoform was not observed in this plant. Expression of Cu/Zn-SOD and Mn-SOD was preferentially enhanced by chilling stress. Expression of Mn-SOD-2 and -4 was enhanced after 48 h of the poststress period. Five APX isoforms were presented in the leaves of cucumber. The intensities of APX-4 and -5 were enhanced by chilling stress, whereas that of APX-3 was significantly increased in the poststress periods after chilling stress. Gel stained for GR activity revealed six isoforms in the plant. Activation levels for most of GR isoforms were higher in the stressed-plants than the control and poststressed-plants, but that of GR-1 isoform was significantly higher in the poststressed-plants than chilling stressed-plants. These results collectively suggest that chilling stress activates the enzymes of an SOD/ascorbate-glutathione cycle under catalase deactivation in the leaves of cucumber, but the response timing of enzyme isoforms against various environmental stresses is not the same for all isoforms of antioxidant enzymes. PMID- 11011096 TI - Analysis of grape ESTs: global gene expression patterns in leaf and berry. AB - Analysis of 2479 ESTs from Vitis vinifera berry tissue and 2438 from leaf revealed that 1% of the ESTs match to known Vitis proteins, 72% to plant proteins, 11% to non-plant, and 16% had no match (P[N]>0.5). The levels of redundancy were similar in the leaf and berry libraries. Only 12% of the genes matched by the ESTs were common to both libraries indicating marked differences in the genes expressed in the two tissues. The abundance of transcripts with predicted cellular roles in leaf and berry were estimated by classifying the primary BLAST matches to known proteins (score >80) into functional categories. Thirty-six percent of the leaf transcripts were involved in photosynthesis, compared to 3% in the berry. This is a much higher proportion of transcripts involved with a function limited to specialized cells, than was found when transcripts of 33 human tissues were compared using a similar approach, suggesting plant cells may involve their cellular machinery to a greater extent in specialized activities than animal cells. Relatively enhanced expression of specific transcription factors, and genes involved in defense, detoxification, stress response, proteolysis, trafficing, and signal transduction, suggests berry tissue is actively engaged in responding to environmental stimuli. PMID- 11011097 TI - Pepper gene encoding a basic beta-1,3-glucanase is differentially expressed in pepper tissues upon pathogen infection and ethephon or methyl jasmonate treatment. AB - A basic beta-1,3-glucanase cDNA clone (CABGLU) was isolated from the cDNA library constructed from hypersensitive response lesions of pepper leaves infected with avirulent strain of Xanthomonas campestris pv. vesicatoria. The deduced polypeptide of CABGLU which contains a C-terminal extension N-glycosylated at a single site characterized as typical structure of class I beta-1,3-glucanase has a high level of identity with tobacco basic beta-1,3-glucanase (77.4%), but only a moderate level of identity with tomato acidic beta-1,3-glucanase (42.6%). Genomic DNA gel blot analysis indicates that the pepper genome contains one or two beta-1,3-glucanase copy genes. Transcripts of the CABGLU gene were more induced in incompatible interactions than in compatible interactions, when inoculated with X. campestris pv. vesicatoria or Phytophthora capsici. Accumulation of CABGLU mRNA was strongly induced in pepper leaves by both ethephon and methyl jasmonate. The CABGLU mRNA was constitutively expressed only in the roots of all the plant organs. These data indicate that the basic beta-1,3 glucanase gene may be induced by pathogen attack and abiotic stresses. PMID- 11011098 TI - Photosynthesis and antioxidant enzymes of phyllodes of Acacia mangium. AB - Physiological processes are influenced by environmental factors and plant characteristics. The distribution of photosynthetic capacity of phyllodes of Acacia mangium Willd. seedlings was studied in relation to the in vivo photosystem II (PSII) function, photosynthetic gas exchange, chlorophyll fluorescence and activities of antioxidant enzymes (superoxide dismutase (SOD) and ascorbate peroxidase (APX)) of phyllodes at different positions on seedlings. There was a vertical gradient in photosynthetic capacity of phyllodes along the shoot. Phyllode 1 (at the apex) showed negative carbon uptake at PPFD lower than 400 umol m(-2) s(-1). High photosynthetic capacities, chlorophyll concentrations, DeltaF/F'(m), and q(P) were observed in phyllodes 4, 6 and 8. The high photosynthetic capacities of mature phyllodes could be attributed to the enhanced availability of CO(2) and the high efficiency of PSII in energy absorption and utilization. Total SOD and APX activities (on a dry weight basis) were highest at phyllode 1 and decreased as the phyllodes matured. The high photosynthetic capacity and low respiration loss in mature phyllodes could be important factors, responsible for the rapid establishment and fast growth of A. mangium in reforestation programs. PMID- 11011099 TI - Cerium and lanthanum promote floral initiation and reproductive growth of Arabidopsis thaliana. AB - The effects of cerium and lanthanum on the vegetative growth, floral initiation and reproductive growth of Arabidopsis thaliana were studied. Addition of cerium nitrate (0.5-10 uM) or lanthanum nitrate (0.5-50 uM) to the culture medium significantly increased the lengths of primary roots, but had no significant effects on the number of rosette leaves produced per plant, plant heights and dry weights during the vegetative growth stage (17 days after seed germination). The percentage of plants bolted was significantly increased with the addition of 0.5 10.0 uM cerium nitrate or lanthanum nitrate. The combination of 0.5 uM cerium nitrate and 0.5 uM lanthanum nitrate was found to be most effective on the induction of floral initiation. The height, dry weight and average number of flower numbers of 35-day-old plants growing in media containing cerium nitrate or/and lanthanum nitrate (0.5-10.0 uM) were found to be significantly higher than those in the control medium. The endogenous levels of cytokinins (zeatin riboside, dihydrozeatin riboside and isopentenyl adenosine) and carbohydrates (sucrose, glucose and fructose) in leaf and root tissues of plants growing in the medium supplemented with 0.5 uM cerium nitrate and 0.5 uM lanthanum nitrate were not significantly different from those of plants in the control medium. Application of 0.5 uM cerium nitrate and 0.5 uM lanthanum nitrate enhanced the effects of 10(-6) M IPA on root growth, plant height and flowering. The role of cerium and lanthanum in promoting floral initiation and reproductive growth and the possibility of developing non-hormonal flowering promoting agents are discussed. PMID- 11011100 TI - Changes in amino acid composition and nitrogen metabolizing enzymes in ripening fruits of Lycopersicon esculentum Mill. AB - The free amino acid content of tomato (Lycopersicon esculentum Mill.) fruits from cultivars Platense, Vollendung and Cherry were determined during ripening. It was found that glutamate markedly increased in red fruits of the three cultivars under study. At this stage, the cv Cherry had the highest relative glutamate molar content (52%) of all the analyzed tomato fruit cultivars. Measurements of nitrogen-assimilating enzyme activities of these fruits showed a decrease in glutamine synthetase (GS, EC 6.3.1.2) during fruit ripening and a concomitant increase in NADH-glutamate dehydrogenase (GDH, EC 1.4.1.3) and aspartate aminotransferase (EC 2.6.1.1) activities. Western blot analysis of protein extracts revealed that while GS was principally present in green fruit extracts, GDH was almost exclusively observed in the extracts of red fruits. These results suggest a reciprocal pattern of induction between GS and GDH during tomato fruit ripening. PMID- 11011101 TI - Characterization of salt-induced changes in gene expression in tomato (Lycopersicon esculentum) roots and the role played by abscisic acid. AB - Examination of tomato (Lycopersicon esculentum Mill) root mRNA profiles by differential display-polymerase chain reaction (DD-PCR) revealed that a salt treatment induced, promoted or repressed the expression of a number of genes. The majority of the observed changes were indicative of a rapid and transient salt induced alteration in gene expression. Twenty partial cDNAs corresponding primarily to salt-induced or up-regulated mRNAs were subsequently cloned and sequenced. The role of abscisic acid (ABA) in regulating salt-responsive gene expression in roots was explored. The DD-PCR data indicate that the majority of the salt-induced changes in the root mRNA profile occurred in an ABA-independent manner. The expression of genes corresponding to six cDNAs was shown unequivocally to be responsive to a salt treatment by RNA blot hybridization. Just two of these were responsive to exogenous ABA and, in salt-treated roots of the ABA-deficient mutant flacca, all were expressed to a level comparable to that in the wild-type. The identity of two of the salt-responsive partial cDNAs is known. The deduced amino acid sequence of one was similar to that of laccases that polymerize a variety of substrates to form resilient structures within the cell wall. One other shared amino acid sequence similarity with the C-terminus of a tobacco pathogen-induced oxygenase (PIOX). It is possible that the PIOX is involved in generating signaling molecules that mediate a general stress response. PMID- 11011102 TI - Significance of sulfhydryl groups in the activity of urease from pigeonpea (Cajanus cajan L.) seeds. AB - Titration of urease from pigeonpea (Cajanus cajan L.), a hexameric protein (mol. wt. 480000; subunit mol. wt. 80000), with 5,5'-dithiobis-(2-nitrobenzoate) (DTNB) reveals the presence of 5.82+/-0.13 'accessible' sulfhydryl groups per molecule of the enzyme protein (i.e. about one 'accessible' SH group per subunit). Denatured enzyme was found to titrate for 12.1+/-0.1 SH groups per molecule (i.e. about two SH groups per subunit). Half of the 'accessible' groups react faster than the remaining at pH 8.5 as well as pH 7.5. However, the reaction was slower at pH 7.5 than 8.5. Time-dependent loss of enzyme activity with DTNB was also found to be biphasic. The enzyme was inactivated at low concentration of p chloromercuribenzoate (p-CMB), N-ethyl maleimide (NEM) and iodoacetamide. The inactivation reactions were biphasic, with half of the activity lost more rapidly than the remaining half. The loss of activity with p-CMB was linearly related to the blocking of accessible SH groups. Inactivation by p-CMB is largely reversible by addition of excess of cysteine. Fluoride ion strongly protects the enzyme against NEM inactivation, however, substrate urea provides much weaker protection against SH group reagents. The significance of these results is discussed. PMID- 11011103 TI - Redox state and peroxidase system in sunflower plants exposed to ozone. AB - Sunflower plants subjected to a short-term fumigation with O(3) (150 ppb for 4 h repeated for 4 days) exhibited an increase in total ascorbate content, accompanied by a marked oxidation of ascorbate, leading to a decrease in its redox state, either at intracellular or extracellular level. O(3) exposure induced a rise in free extracellular peroxidase (POD) activity, assayed by syringaldazine as electron donor, as well as in the ionically and covalently cell wall bound PODs. On the contrary, the activity of both extracellular and intracellular guaiacol-POD did not show significant changes as a consequence of the pollutant exposure. The stimulation of syringaldazine-POD activities may be related to the effect of ozone on the growth of the cells, inducing an early senescence through the activation or acceleration of lignification processes. Beside, the reduced plasticity of the cell wall may oppose an unspecific mechanical resistance against the abiotic stress induced by the ozone exposure. PMID- 11011104 TI - A fuzzy-neural system for identification of species-specific alarm calls of Gunnison's prairie dogs. AB - In this study we describe the design and application of an automated classification system that utilizes artificial intelligence to corroborate the finding that Gunnison's prairie dogs have different alarm calls for different species of predators. This corroboration is strong because it utilizes an entirely different analysis technique than that used in the original research by Slobodchikoff et al. [Slobodchikoff, C.N., Fischer, C., Shapiro, J., 1986. Predator-specific alarm calls of prairie dogs. Am. Zool. 26, 557] or in subsequent study done by Slobodchikoff et al. [Slobodchikoff, C.N., Kiriazis, J., Fischer, C., Creef, E., 1991. Semantic information distinguishing individual predators in the alarm calls of Gunnison's prairie dogs. Anim. Behav. 42, 713 719]. The study described here also is more completely automated than earlier study in this area. This automation allowed a large volume of field data to be processed where all measurements of relevant parameters were performed through software control. Previous study processed a smaller data set and utilized manual measurement techniques. The new classification system, which combines fuzzy logic and an artificial neural network, classified alarm calls correctly according to the eliciting predator species, achieving accuracy levels ranging from 78.6 to 96.3% on raw field data digitized with low quality audio equipment. PMID- 11011105 TI - Rats more readily acquire a time-of-day go no-go discrimination than a time-of day choice discrimination. AB - Male Sprague-Dawley rats were used to compare six time-place training procedures that differed with respect to housing or training conditions. All procedures involved training food-deprived rats to enter one choice arm of a T-maze during a morning test session and to enter the other choice arm during an afternoon session to obtain Cocoa Puffs(R). The task proved to be difficult. Only 39 of 49 rats attained a criterion of nine correct choices on ten consecutive trials within a total of 120 trials. Making one choice arm distinct, limiting consecutive same correct choices to two, giving one session during the light and one during the dark portion of the light cycle, extinguishing perseveration of the same choice responses, and housing the rats with a natural light cycle all failed to significantly decrease the errors or trials to a 90% correct choice criterion. In contrast, all responding rats (n=7) showed significantly better than chance performance within 48 trials when the task was a go no-go discrimination based on time of day. Learning to make a response during a session when a choice to either choice arm is reinforced and to withhold responding during a session when a choice to neither choice arm is reinforced was relatively easy for the rats to acquire. Continued high level performance after the light cycle was eliminated, a random feeding schedule was initiated, and other time related cues were masked suggests that the rats used internal cues or an internal clock to make the correct go or no-go response. It was concluded that rats are prepared to use time of day as an occasion-setting stimulus, but have difficulty using time of day as a signal for a specific response. PMID- 11011106 TI - Prey catching in the archer fish: does the fish use a learned correction for refraction? AB - An answer to the question of how the archer fish hits an aerial insect, despite the refraction of light at the surface of the water has not yet been found. The aims of the present studies are to find out: (1) whether the fish applies a learned correction with the virtual image as a point of reference; (2) whether deprivation of practice in squirting affects performance. For the first aim the accuracy of squirts was measured in 30 subjects. Contrary to suggestions from the literature, elevation failures were prominent but the frequencies of over- and under-squirting did not differ, which does not support the idea that the fishes applied a learned correction for refraction by using feedback from the efficacy of squirts. For the second aim, five experimental subjects were deprived of practice, whereas six control subjects got daily practice, during 6 months. The only significant difference, found thereafter, was that during the first session experimental subjects aimed more often before squirting than control subjects did, but hitting was not affected. A number of subjects developed abnormal mandibles which inevitably led to squirting too high. Our findings do not support the hypothesis that the archer fish uses learned corrections for refraction. PMID- 11011107 TI - Influence of gender and behavioural lateralisation on two exploratory models of anxiety in C3H mice. AB - Behavioural lateralisation, which has been postulated to be an individual personality trait, is related to the activity of various physiological systems including the immune system. As lateralisation has been related to anxiety, which is known to influence immune reactivity, it can be hypothesized that the relation between lateralisation and immune reactivity involves individual behavioural patterns as they appear in exploratory-based anxiety models. In order to answer this question, a behavioural investigation focussing on exploratory activity was undertaken in male and female C3H mice previously selected for their paw preference. The observations were performed using two generic paradigms: elevated plus-maze and open field. Exploratory behaviour in the open field, but not in the plus-maze, was influenced by the interactive effect of gender and behavioural lateralisation. A significant difference between male and female mice was found in left-pawed but not in right-pawed nor ambidextrous animals, left-pawed female mice displaying the less exploratory behaviours. These results provide a first evidence of inter-individual variations in exploratory behaviours involving interaction between gender and lateralisation. PMID- 11011108 TI - Time estimation by pigeons on a fixed interval: the effect of pre-feeding. AB - Pigeons well trained on a fixed interval 10-s schedule of reinforcement were tested on the peak procedure. In a successive conditions design, they were either pre-fed or not in the experimental setting. Pre-feeding decreased the rate of responding. It also led to a maximum rate of responding that occurred 2-3 s later than in the control condition, where the maximum occurred at the usual time of reinforcement. The shift in peak time in response to pre-feeding shows that peak time may not be a pure measure of timing. The results are also interpreted in terms of timing theories. PMID- 11011109 TI - Effects of auditory stimuli correlated with different probabilities of water delivery in a limited-hold temporal schedule. AB - Four rats were exposed to two different tone frequencies, each tone being correlated with independent probabilities of water delivery in a temporally defined schedule. The schedule consisted of a 60-s T cycle, with 30-s t(D) and t(Delta) succesive subcycles. t(D) and t(Delta) were assigned complementary probabilities of water delivery (1.0-0, 0.75-0.25, 0.5-0.5 and 1.0-0) in succesive phases. Each of the first three experimental phases was followed by five consecutive probe sessions using the same probability values, and in which water was delivered independently of responding at the end of each subcycle. Three additional rats were exposed to the same conditions, except that only one tone was used in t(D) and t(Delta). The highest frequencies of responding were observed in two of the rats exposed to differential tone frequencies, an effect that was correlated with lower percentages of lost water deliveries, shorter response latencies, and less variability in the intervals between successive water presentations. In the last phase in which probabilities of water delivery in t(D) and t(Delta) were 1.0 and 0, respectively, response frequency was always higher in t(Delta). These results suggest that the number of water deliveries obtained in the early cycles of every session were responsible for the total number of responses and its correlated effects. PMID- 11011110 TI - An evidence-based approach to prescribing NSAIDs in the treatment of osteoarthritis and rheumatoid arthritis: The Second Canadian Consensus Conference. AB - The Second Canadian Consensus Conference was convened to discuss the latest developments in the management of osteoarthritis (OA) and rheumatoid arthritis (RA), and to make evidence-based recommendations, specifically regarding the use of nonsteroidal anti-inflammatory drugs (NSAIDs) for these indications in primary care practice. The recent availability of cyclo-oxygenase-2-specific inhibitors has raised questions as to their role in the pharmacological management of OA and RA, particularly in relation to conventional treatments such as acetaminophen and nonspecific NSAIDs (with or without misoprostol or proton pump inhibitors). The recommendations in this document, which were arrived at through critical review of data from published randomized, clinical trials, deal with treatments of choice, information to discuss with patients, use of NSAIDs in patients at risk for serious upper gastrointestinal complications, renal or hepatic impairment or congestive heart failure, appropriate follow-up, and the use of NSAIDs with anti- hypertensives, warfarin, low dose acetylsalicylic acid and other medications. The goal of these recommendations is to improve patient outcomes in the primary care setting by maximizing treatment efficacy and minimizing rates of adverse events. PMID- 11011111 TI - Polymerase chain reaction with confronting two-pair primers for polymorphism genotyping. AB - A novel PCR method using confronting two-pair primers, named PCR-CTPP, is introduced to detect a single nucleotide polymorphism (base X or Y). One primer for the X allele is set to include X' at the 3' end (antisense), where X' is the antisense of X, with the counterpart sense primer upstream. For the Y allele, a sense primer including Y at the 3' end is set, with the antisense primer downstream. One common band and one specific band for each allele are amplified, which allows genotyping directly by electrophoresis. This method is exemplified by application to the polymorphisms of beta-adrenoceptor 2 and interleukin 1B. It is simpler than PCR-RFLP (restriction fragment length polymorphism), which requires incubation with a restriction enzyme, and is suitable for genotyping in studies of genetic epidemiology involving hundreds of samples. PMID- 11011112 TI - A novel gene "Niban" upregulated in renal carcinogenesis: cloning by the cDNA amplified fragment length polymorphism approach. AB - A modified AFLP (amplified fragment length polymorphism) method was employed to isolate genes differentially expressed in renal carcinogenesis of Tsc2 gene mutant (Eker) rats. One gene, selected for further investigation, was named "Niban" "second" in Japanese), because it is the second new gene to be found after Erc (expressed in renal carcinoma) in our laboratory. Importantly, "Niban" is well expressed even in small primary rat Eker renal tumors, more than in progressed cell lines, and is also expressed in human renal carcinoma cells, but not in normal human or rat kidneys. Chromosome assignment was to RNO 13 in the rat, and HSA 1. This "Niban" gene is a candidate as a marker for renal tumor, especially early-stage renal carcinogenesis. PMID- 11011113 TI - Absence of germline CHK2 mutations in familial gastric cancer. AB - Recently, the CHK2 gene was identified as being a candidate gene responsible for Li-Fraumeni syndrome (LFS). Gastric cancer is often clustered in families with LFS, so it is possible that germline CHK2 mutation is also present in familial gastric cancer (FGC). We therefore defined the genomic structure of the CHK2 gene, designed intronic primers, and searched for germline CHK2 mutations in 25 FGC cases by polymerase chain reaction-single strand conformational polymorphism analysis of the entire coding region. In all of the 25 cases, at least two siblings had histories of gastric cancer. There were no FGC cases that showed germline CHK2 mutations. Thus, it was indicated that germline CHK2 mutations do not contribute to the familial clustering of gastric cancer. PMID- 11011114 TI - No association of the 5' promoter region polymorphism of CYP17 with breast cancer risk in Japan. AB - To examine the association between breast cancer risk and a T-to-C substitution polymorphism at the 5' promoter region of CYP17, a case-control study was conducted at Aichi Cancer Center Hospital in Japan. Subjects were 144 histologically confirmed breast cancer patients diagnosed in the past 4 years and 166 hospital controls without cancer. Allele frequency among controls was 44.9% (95% confidence interval; 39.5 - 50.2) for C allele. Odds ratio (OR) of the polymorphism relative to TT-genotype was 0.97 (0.58 - 1.64) for TC-genotype and 0.81 (0.39 - 1.68) for CC-genotype. Subgroup analyses revealed that the OR was not statistically significant for the subgroups stratified by interval after diagnosis, age at menarche, age at first birth, menopausal status, body mass index, and mother / sisters' history of breast cancer. Consistent with previous studies conducted in other countries, the 5' promoter region polymorphism of CYP17 affected breast cancer risk of Japanese women to a limited extent. Although this is not a large-scale case-control study with population controls, these findings provide enough information to discourage further studies on the association between this polymorphism and breast cancer risk in Japan at large, and suggest that this polymorphism is useless for breast cancer risk estimation. PMID- 11011115 TI - Chemoprevention by nimesulide, a selective cyclooxygenase-2 inhibitor, of 2-amino 1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced mammary gland carcinogenesis in rats. AB - Breast cancer is common in women all over the world, and exploration of chemopreventive approaches to this cancer is very important. Nimesulide, a selective inhibitor of cyclooxygenase-2 (COX-2), is a good candidate as a chemopreventive agent with low toxicity. We examined its effects on mammary tumor development in female Sprague-Dawley rats induced with the environmental carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Rats at 7 weeks of age received intragastric intubations of PhIP (85 mg / kg body weight) 4 times weekly for 2 weeks and were maintained on control diet (high fat diet) or experimental diet (high fat diet supplemented with 400 ppm nimesulide) throughout the experiment. COX-2 protein was over-expressed in epithelial cancer cells and stromal cells of the PhIP-induced mammary carcinomas, but was weak or not apparent in normal mammary gland cells. The development of mammary carcinomas was clearly suppressed by administration of nimesulide. The carcinoma incidence was 51% as compared to 71% for the control diet group. The average multiplicity of carcinomas in the experimental diet group was 1.2 +/- 0.2 (P < 0.05), significantly smaller than the control diet group value (2.6 +/- 0. 5). The size of carcinomas was also clearly decreased; 1.1 +/- 0.4 cm(3) / rat in experimental diet group (P < 0.05), 4.1 +/- 1.3 cm(3) / rat in the control diet group. The results therefore provide evidence that the selective COX-2 inhibitor, nimesulide, possesses chemopreventive activity against PhIP-induced mammary carcinogenesis in rats. PMID- 11011116 TI - Chemopreventive effect of curcumin on N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in rats. AB - Modifying effects of curcumin (derived from the rhizome of Curcuma longa L.) during the initiation or post-initiation phase of N-nitrosomethylbenzylamine (NMBA)-induced esophageal carcinogenesis were investigated in male F344 rats. Five-week-old rats were divided into 5 groups, and groups 1, 2 and 3 were given intraperitoneal injections of NMBA (0.5 mg / kg body weight / injection 15 times) for 5 weeks from 7 weeks old to induce esophageal neoplasms. Groups 2 and 3 were fed the diet containing 500 ppm curcumin during the initiation and post initiation phases, respectively. Group 4 was given the diet containing curcumin throughout the experiment, and group 5 was kept on the basal diet alone and served as an untreated control. Incidence and multiplicity of esophageal neoplasms of group 1 (NMBA alone) were 66.7% and 0.83 +/- 0.70, respectively. Those of groups 2 and 3 were significantly less than those of group 1 (39.3%, 0.46 +/- 0.64, P < 0.05; 33.3%, 0.36 +/- 0.56, P < 0.05, respectively). Furthermore, the incidence and multiplicity of esophageal preneoplastic lesions (moderate or severe epithelial dysplasia) of group 2 (57.1%, 0.61 +/- 0.57; 40%, 0.29 +/- 0.46) or 3 (56.7%, 0.67 +/- 0.66; 23.3%, 0.23 +/- 0.43) were less than those of group 1 (100%, 1.67 +/- 0.70; 70.8%, 0.92 +/- 0.72) (P < 0.05). In this experiment, feeding of curcumin significantly decreased the expression of cell proliferation biomarkers (5-bromo-2'-deoxyuridine labeling index) in the non lesional esophageal epithelium (P < 0.01). These findings indicate that curcumin inhibits NMBA-induced esophageal carcinogenesis when given during the post initiation as well as initiation phase. This inhibition may be related to suppression of the increased cell proliferation induced by NMBA in the esophageal epithelium. PMID- 11011118 TI - Co-expression of thymidine phosphorylase and heme oxygenase-1 in macrophages in human malignant vertical growth melanomas. AB - Expression of thymidine phosphorylase (TP) is often associated with tumor angiogenesis and / or prognosis in patients. Further, infiltration of macrophages is closely correlated with the depth of tumor and angiogenesis in melanomas. In this study, we examined the expression of TP and an activated macrophage-specific enzyme, heme oxygenase-1 (HO-1), involved in malignancy in 22 cases with melanomas. TP was strongly expressed not only in CD68-positive macrophages in and around tumors, but also in S100 protein-positive melanoma cells, fibroblasts and keratinocytes. By contrast, HO-1 was specifically expressed in macrophages, but only slightly in melanoma cells and other cell types in the stroma of melanomas. We thus observed apparent co-expression of TP and HO-1 in macrophages infiltrating in the late stage of malignant melanomas. There appeared increasing numbers of TP-positive cells in Clark level IV and V melanoma compared with Clark level I (in situ) melanoma, and there was also a close correlation between numbers of TP-positive cells and HO-1-positive cells. Both TP- and HO-1-positive macrophages could be observed in the stroma in and around tumors in vertical growth melanomas. PMID- 11011117 TI - Lack of modifying effects of environmental estrogenic compounds on the development of thyroid proliferative lesions in male rats pretreated with N-bis(2 hydroxypropyl)nitrosamine (DHPN). AB - The modifying effects of various environmental estrogenic compounds on thyroid carcinogenesis were investigated in a rodent two-stage carcinogenesis model. The compounds examined were a soy isoflavone mixture (SI) and genistein (GEN) as phytoestrogens, nonylphenol (NP) as a xenoestrogen, 3-chloro-4-(dichloromethyl)-5 hydroxy-2(5H)-furanone (MX) as a thyroid carcinogen and sulfadimethoxine (SDM) as a known thyroid tumor promoter. Five-week-old male F344 rats were given a single subcutaneous injection of N-bis(2-hydroxypropyl)nitrosamine (DHPN; 2800 mg / kg, body weight) or the vehicle alone. Starting one week thereafter, GEN (250 or 25 ppm in diet), SI (400 ppm in diet), NP (250 or 25 ppm in diet), MX (30 ppm, in drinking water) or SDM (1000 ppm in drinking water) was administered for 12 weeks. Major organs including the thyroid, pituitary, liver, kidney, testis, brain and pancreas were weighed and histopathological observation was performed. Thyroid weights were significantly increased (P < 0.001) only in the SDM treatment groups, especially with DHPN pretreatment. Kidney weights were slightly increased in the NP or MX treatment groups, albeit without statistical significance. Histopathologically, thyroid proliferative lesions were only observed in the SDM alone or DHPN + SDM group with significant focal hyperplasias, adenomas and adenocarcinomas limited to the combined treatment case. There were no organ weight changes or histopathological lesions in the major organs including the thyroid in the GEN, SI, NP, and MX treatment groups regardless of DHPN pretreatment. Our results thus indicate that the weakly estrogenic compounds GEN, SI and NP and the environmental rat thyroid carcinogen MX do not exert any modifying effects on thyroid carcinogenesis in rats under the present experimental conditions. PMID- 11011119 TI - Regulation of granulocyte colony-stimulating factor and parathyroid hormone related protein production in lung carcinoma cell line OKa-C-1. AB - Previously we have established a clonal squamous cell carcinoma cell line OKa-C-1 derived from lung cancer of a patient with marked leukocytosis and hypercalcemia. OKa-C-1 cells simultaneously produce granulocyte colony-stimulating factor (G CSF) and parathyroid hormone-related protein (PTHrP) at the single cell level and cause paraneoplastic syndromes in nude mice bearing the tumor. It is known that the production of G-CSF and PTHrP is individually regulated by inflammatory cytokines in various malignant cells. To investigate the common factors in the regulation of G-CSF and PTHrP production in OKa-C-1 cells, we examined the effects of some inflammatory agents [lipopolysaccharide (LPS), phorbol-12 myristate-13-acetate (PMA), tumor necrosis factor-alpha (TNF-alpha), interleukin 1 (IL-1) beta and IL-6] on G-CSF and PTHrP production, by means of enzyme-linked immunosorbent assay (ELISA), immunoradiometric assay (IRMA) and quantitative reverse transcription-polymerase chain reaction (RT-PCR). TNF-alpha or IL-1beta induced both G-CSF and PTHrP production in the conditioned medium. TNF-alpha synergized with IL-1beta to significantly increase G-CSF production. In addition, TNF-alpha and IL-1beta strongly induced G-CSF mRNA with peaks at 2 and 6 h respectively. Although PTHrP production was also strongly induced by TNF-a PTHrP mRNA expression was more strongly induced by PMA than by TNF-alpha. Thus, TNF alpha and IL-1beta could be common factors that individually and synergistically regulate G-CSF and PTHrP production in OKa-C-1 cells. Moreover, G-CSF and PTHrP production could be not only transcriptionally, but also posttranscriptionally regulated by other factors. PMID- 11011120 TI - Prognostic significance of occult bone marrow micrometastases of breast cancer detected by quantitative polymerase chain reaction for cytokeratin 19 mRNA. AB - Amplification of cytokeratin 19 (CK19) transcripts by reverse transcriptase polymerase chain reaction (RT-PCR) has been shown to be a highly sensitive assay for the detection of bone marrow micrometastases (BMM) of breast cancer, but recent studies have demonstrated the occurrence of false-positive results due to low-level, illegitimately transcribed CK19 in normal bone marrow tissue. One approach to solve this problem is to develop a quantitative CK19 RT-PCR assay and to introduce a cut-off value which can distinguish between illegitimate expression and cancer-specific expression levels. In the present paper, we describe a quantitative CK19 RT-PCR assay using a real-time automated PCR system. The number of CK19 transcripts was normalized to that of GAPDH transcripts as an internal control for quality and quantity of cDNA. The cut-off value for the ratio of CK19 to GAPDH transcripts was set at 10(-4) since the ratio never exceeded this value in the control bone marrow samples (n = 12). In total, 117 bone marrow aspirates from stage I - III patients with invasive breast cancers were subjected to CK19 RT-PCR assay and immunocytological examination. Forty (34.2%) were found to be BMM-positive by CK19 RT-PCR assay whereas only three (2.6%) were found to be BMM-positive by immunocytology. Multivariate analysis has shown that occult BMM detected by CK19 RT-PCR is a significant risk factor for relapse, being independent of axillary lymph node metastases. PMID- 11011121 TI - ICAM-1 (intercellular adhesion molecule-1) gene transfection inhibits lymph node metastasis by human gastric cancer cells. AB - Lymph node metastasis is one of the prognostic factors in gastric cancer. We have previously reported that decreased intercellular adhesion molecule-1 (ICAM-1) expression on cancer cells is associated with lymph node metastasis using a gastric cancer cell. In this study, we transfected ICAM-1 gene into a gastric cancer cell line, 2MLN, and analyzed the effect on lymph node metastasis in vitro and in vivo. A significantly greater amount of peripheral blood mononuclear cells (PBMC) adhered to ICAM-1 transfected 2MLN cells, 2MLN / ICAM cells, than to 2MLN / Vector cells. The lysis of 2MLN / ICAM cells by PBMC was significantly increased compared with that of 2MLN / Vector cells. The tumor growth rate of 2MLN / ICAM cells was significantly decreased in vivo. Lymph node metastases caused by 2MLN / ICAM cells were recognized as being fewer in number and smaller, while many lymph node metastases were caused by 2MLN cells. Histologic findings showed that leukocytes were heavily infiltrated in both the 2MLN / ICAM tumors and metastatic lesions, while only a few leukocytes were observed in the lesions associated with 2MLN cells. The above findings indicate that ICAM-1 gene transduction could prove to be an effective gene therapy for lymph node metastasis of gastric cancer. PMID- 11011122 TI - Modulation of myogenic differentiation in a human rhabdomyosarcoma cell line by a new derivative of 5-fluorouracil (QF-3602). AB - The in vitro study of mechanisms involved in drug-induced maturation has made it possible to use differentiation-based therapy in clinical practice. The goal of this new therapy is the development of specific agents to induce cancer cells to stop proliferating and express characteristics of normal cells. Recently, by structural modifications of 5-fluorouracil (5-FU), we synthesized a new pyrimidine acyclonucleoside-like compound, 1-?[3-(3-chloro-2-hydroxypropoxy)-1 methoxy]propyl?-5-fluorouracil (QF-3602), which showed in rhabdomyosarcoma cells a low toxicity and time-dependent growth inhibition. In this work, we compared the degree of myogenic differentiation of RD rhabdomyosarcoma (RMS) cells after treatment with QF-3602 and 5-FU. Scanning and transmission electron microscopy (SEM and TEM) and immunocytochemical analyses showed that QF-3602 induced the appearance of myofilaments along the myotube-like giant RD cells, an increase in fibronectin and a decrease in vimentin expression. In contrast, only minor changes were observed with 5-FU. Moreover, polymerase chain reaction (PCR) analyses showed that QF-3602 did not induce overexpression of the mdr 1 gene, a resistance mechanism that frequently appears in classical cytotoxic therapy in these tumors. Compounds obtained by structural modifications of 5-FU may be useful in differentiation therapy as a new approach to the treatment of RMS. PMID- 11011123 TI - p53 mutations in prostatic intraepithelial neoplasia and concurrent carcinoma: analysis of laser capture microdissected specimens from non-transition and transition zones. AB - Prostatic intraepithelial neoplasia (PIN) is characterized by intraluminal proliferation of epithelial cells and is divided into high-grade (HGPIN) and low grade (LGPIN) lesions. HGPIN is regarded as the most likely precursor of prostatic cancer (PCA). Microdissected DNA selectively extracted from paraffin embedded sections of 27 cases with PCA were analyzed for p53 mutation in exons 5 8 by single-strand conformation polymorphism of polymerase chain reaction amplified DNA fragments (PCR-SSCP) followed by direct sequencing. These patients received total prostatectomy (27 cases). After a review of histologic sections, DNA was extracted from 193 locations; 111 lesions from 27 cases with HGPIN (75 lesions from non-transition zone and 36 from transition zone), 55 lesions from 27 cases with PCA (30 lesions from non-transition zone and 25 from transition zone), and 27 from 27 benign glands. Analysis revealed 27 mutations of the p53 gene in 24 lesions from 12 cases. Benign glands adjoining PIN and / or PCA had no mutations. All mutations were point mutations: 17 missense, 7 silent, and 2 nonsense. Mutations were detected in 6 cases (22.2%) or 13 of 111 lesions (11.7%) with HGPIN and 8 cases (29.6%) or 11 of 55 lesions (20.0%) with PCA. In a given case, HGPIN and PCA lesions had different p53 mutations from each other, suggesting multiclonal development of prostatic precancerous lesions. The frequency of p53 mutation of PCA was significantly higher in the non-transition zone (33.3%) than in the transition zone (4%), and higher in the stage T3 cases (30.3%) than in the stage T2 cases (4.5%, 1 of 22 lesions) (both P < 0.05). Frequency of p53 mutation of PIN in the non-transition zone (14.7%) was higher than that in the transition zone (5.6%), although the difference was not significant. The frequency rate of p53 mutation in HGPIN close to PCA ( 2 mm from PCA (3%). All these findings indicate that the p53 gene mutations are involved in prostatic carcinogenesis and explain why the non-transition zone is the predominant site of PCA. PMID- 11011124 TI - Relations of insulin resistance and serum concentrations of estradiol and sex hormone-binding globulin to potential breast cancer risk factors. AB - There is a hypothesis that hyperinsulinemia or insulin resistance may be a mediator for breast cancer risk factors. On the other hand, some, but not all, of the well-known risk factors of breast cancer have been associated with serum estrogen concentrations. We assessed the relationships of potential breast cancer risk factors to indicators of insulin resistance, fasting plasma insulin concentration and homeostasis model assessment insulin resistance (HOMA-R), in 88 postmenopausal Japanese women. We also examined whether insulin resistance would explain the association of breast cancer risk factors with serum estradiol and sex hormone-binding globulin (SHBG). Information on potential breast cancer risk factors, such as demographic characteristics, smoking and drinking habits, diet, exercise, menstrual and reproductive factors, was obtained by self-administered health questionnaire including a validated semiquantitative food frequency questionnaire. Body mass index (BMI) was significantly correlated with the ratio of estradiol to SHBG (Spearman r = 0.30, P = 0.0004), fasting plasma insulin (r = 0.45) and HOMA-R (r = 0.43, P = 0.0001) after controlling for age. The correlations were still significant between BMI and estradiol / SHBG ratio (r = 0.21, P = 0.047) after controlling for fasting plasma insulin and between BMI and fasting plasma insulin (r = 0.40, P = 0. 0001) as well as HOMA-R (r = 0.38, P = 0.0003) after controlling for estradiol / SHBG ratio. There is a possibility that effect of BMI on breast cancer risk is mediated by both insulin resistance and estrogen metabolism. PMID- 11011136 TI - Metabolism of palmitic and docosahexaenoic acids in Reuber H35 hepatoma cells. AB - In this work, we have modified the fatty acid composition of Reuber H35 hepatoma cells by supplementation of the culture medium with a saturated (palmitic) or a polyunsaturated (docosahexaenoic) acid. These fatty acids were incorporated into total lipids and phospholipids of hepatoma cells. Palmitic acid readily increased the percentage of its monounsaturated derivative (16:1 n-7). When both fatty acids were supplemented at the same concentration, the percentage of docosahexaenoic acid in the total lipids and phospholipids of Reuber H35 cells increased more than that of palmitic acid. Although the levels of 16:0 increased, the addition of docosahexaenoic acid to the culture medium decreased the percentages of monoenoic acids. From our results, it can be concluded that palmitic and docosahexaenoic acids modify the fatty acid composition of Reuber H35 hepatoma cells. The profound changes induced by docosahexaenoic acid, especially those in the phospholipid fraction, may be of great interest given the main role of these components in the regulation of chemical and physical properties of biological membranes and/or membrane systems. PMID- 11011137 TI - Binding of an intrinsic ATPase inhibitor to the F(1)FoATPase in phosphorylating conditions of yeast mitochondria. AB - Yeast mitochondrial ATP synthase has three regulatory proteins; ATPase inhibitor, 9K protein, and 15K protein. A mutant yeast lacking these three regulatory factors was constructed by gene disruption. Rates of ATP synthesis of both wild type and the mutant yeast mitochondria decreased with decrease of respiration, while their membrane potential was maintained at 170-160 mV under various respiration rates. When mitochondrial respiration was blocked by antimycin A, the membrane potential of both types of mitochondria was maintained at about 160 mV by ATP hydrolysis. ATP hydrolyzing activity of F(1)FoATPase solubilized from normal mitochondria decreased in proportion to the rate of ATP synthesis, while the activity of the mutant F(1)FoATPase was constant regardless of changes in the rate of phosphorylation. These observations strongly suggest that F(1)FoATPase in the phosphorylating mitochondria is a mixture of two types of enzyme, phosphorylating and non-phosphorylating enzymes, whose ratio is determined by the rate of respiration and that the ATPase inhibitor binds preferentially to the non phosphorylating enzyme. PMID- 11011138 TI - EmPLiCS: an empirical approach for structure-based design of natural peptide drugs. AB - The computer implementation of a peptide drug-design strategy has been developed. The system is named EmPLiCS (Empirical Peptide Ligand Construction System) according to the strategy of the system, which searches for peptide-ligand structures by referring to empirical rules that are derived from known protein 3D structures. The system was tested on several known peptide-protein complexes. The results demonstrated the ability of this system to detect key residues of peptides that are crucial for interaction with their specific proteins. The system also showed the ability to detect the main chain trace of these peptides. Some of the main chain atoms were detected even though the complete primary structures were not reproduced, suggesting that main chain structure is important in peptide-protein recognition. The results of the present study demonstrated that the empirical rules-based system can generate significant information for use in the design of natural peptide drugs. PMID- 11011140 TI - Enhanced thermostability of the single-Cys mutant subtilisin E under oxidizing conditions. AB - We obtained enhanced thermostability by replacing Ser161 with Cys in subtilisin E from Bacillus subtilis, a cysteine-free alkaline serine protease. The Ser161Cys mutant subtilisin E was purified from the culture supernatant of the recombinant B. subtilis in an oxidizing environment. SDS-polyacrylamide gel electrophoresis and mass spectrometry under oxidizing conditions indicated that the mutant enzyme in part formed an oligomeric protein, which may contain an intermolecular disulfide bond between two surface Cys residues at position 161. Further, no free sulfhydryl groups were detected in the mutant enzyme, suggesting the sulfhydryl modification in a monomeric form under oxidizing conditions. The Ser161Cys mutant enzyme showed a catalytic efficiency equivalent to that of the wild-type enzyme. The half-life of thermal inactivation of the mutant was found to be 2-4 times longer than that of the wild-type enzyme. The optimum temperature of the mutant was 55 degrees C, which was 5 degrees C higher than that of the wild-type enzyme. Under reducing conditions, however, the characteristics of the mutant enzyme reverted to those of the wild-type enzyme. Similar results were obtained for another Cys mutant as to position 194 (wild-type, Ser), which is the same surface residue as Ser161. Possible reasons for the enhanced thermostability of the single-Cys mutant subtilisins E under oxidizing conditions are discussed in terms of two different mechanisms. PMID- 11011139 TI - Genetic characterization of rbt mutants that enhance basal transcription from core promoters in Saccharomyces cerevisiae. AB - While this Saccharomyces cerevisiae SIN4 gene product is a component of a mediator complex associated with RNA polymerase II, various studies suggest the involvement of Sin4 in the alteration of higher-order chromatin structure. Our previous analysis of a sin4 mutant suggested that the mechanisms of transcriptional repression by Sin4 (mediator) and the Tup1-Ssn6 complex (general repressor) are different. To elucidate the way in which these two repression systems are interrelated, we isolated mutants that exhibit enhanced transcription of a reporter gene harboring the upstream activation sequence (UAS), but still are subject to Tup1-Ssn6-mediated repression. Besides sin4, rgr1, tup1, and ssn6 mutants, we also obtained new mutants that enhance basal transcription even from a core promoter without UAS. Such mutants, designated rbt for regulator of basal transcription, can be classified into at least six complementation groups, i.e., four single (rbt1 to rbt4) and two apparently double (rbt5 rbt6 and rbt7 rbt8) mutations. The phenotype of rbt mutants is dependent on the TATA box and not specific to the integration site or kind of core promoter. No significant difference in micrococcal nuclease (MNase) accessibility to the core promoter of test genes was observed between rbt mutants and the wild-type strain, indicating that the higher-order chromatin structure of the core promoter region is not significantly altered in these mutants. The rbt1 to rbt4 mutations are suppressed by the Dgal11 mutation as in the case of the sin4 mutation, but give rise to a different profile from the sin4 mutation with regard to the activity of some of the promoters. From these observations, we suggest that RBT gene product(s) could be novel mediators that act with or in close association with Sin4 but have a function distinct from that of Sin4. Moreover, the fact that rbt mutations nullify Tup1-Ssn6 general repressor-mediated repression is consistent with the idea that the mechanisms of Rbt (mediator)- and Tup1-Ssn6 (general repressor) mediated repression are interconnected but substantially different. PMID- 11011141 TI - The role of structural intersubunit microheterogeneity in the regulation of the activity in hysteresis of ribulose 1, 5-bisphosphate carboxylase/oxygenase. AB - Many enzymes are composed of subunits with the identical primary structure. It has been believed that the protein structure of these subunits is the same. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) comprises eight large subunits with the identical amino acid sequence and eight small subunits. Rotation of the side chains of the lysine residues, Lys-21 and Lys-305, in each of the eight large subunits in spinach RuBisCO in two ways produces microheterogeneity among the subunits. These structures are stabilized through hydrogen bonds by water molecules incorporated into the large subunits. This may cause different effects upon catalysis and a hysteretic, time-dependent decrease in activity in spinach RuBisCO. Changing the amino acid residues corresponding to Lys-21 and Lys-305 in non-hysteretic Chromatium vinosum RuBisCO to lysine induces hysteresis and increases the catalytic activity from 8.8 to 15.8 per site per second. This rate is approximately five times higher than that of the higher plant enzyme. PMID- 11011142 TI - Mouse T-cell antigen rt6.1 has thiol-dependent NAD glycohydrolase activity. AB - Mouse Rt6.1 and Rt6.2, homologues of rat T-cell RT6 antigens, catalyze arginine specific ADP-ribosylation. Without an added ADP-ribose acceptor, Rt6.2 shows NAD glycohydrolase (NADase) activity. However, Rt6.1 has been reported to be primarily an ADP-ribosyltransferase, but not an NADase. In the present study, we obtained evidence that recombinant Rt6.1 catalyzes NAD glycohydrolysis but only in the presence of DTT. The NADase activity of Rt6.1 observed in the presence of DTT was completely inhibited by N-ethylmaleimide (NEM). Native Rt6.1 antigen, immunoprecipitated from BALB/c mouse splenocytes with polyclonal antibodies generated against recombinant RT6.1, also exhibited NADase activity in the presence of DTT. Compared with Rt6.2, Rt6.1 has two extra cysteine residues at positions 80 and 201. When Cys-80 and Cys-201 in Rt6.1 were replaced with the corresponding residues of Rt6.2, serine and phenylalanine, respectively, Rt6.1 catalyzed the NADase reaction even in the absence of DTT. Conversely, replacing Ser-80 and Phe-201 in Rt6.2 with cysteines, as in Rt6.1, converted the thiol independent Rt6.2 NADase to a thiol-dependent enzyme. Kinetic study of the NADase reaction revealed that the affinity of Rt6.1 for NAD and the rate of catalysis increased in the presence of DTT. Moreover, the NADase activity of Rt6.1 expressed on COS-7 cells was stimulated by culture supernatant from activated mouse macrophages, even in the absence of DTT. From these observations, we conclude that t!he Rt6.1 antigen has thiol-dependent NADase activity, and that Cys-80 and Cys-201 confer thiol sensitivity to Rt6.1 NADase. Our results also suggest that upon the interaction of T-cells expressing Rt6.1 with activated macrophages, the NADase activity of the antigen will be stimulated. PMID- 11011144 TI - Autoantibody activity of IgG rheumatoid factor increases with decreasing levels of galactosylation and sialylation. AB - The occurrence of N-linked oligosaccharides lacking galactose is significantly higher than normal in serum IgG of patients with rheumatoid arthritis (RA) in whom rheumatoid factor (RF), an autoantibody against autologous IgG, has been detected. In the present study, IgGs with and without RF activity (IgGRF and non RF IgG, respectively) were prepared from sera of RA patients, and their oligosaccharide structures were characterized in order to investigate the relationship between RF activity and glycosylation. Three IgGRF fractions and a non-RF IgG fraction were obtained based on their ability to bind to an IgG Sepharose column. The specific RF activity, as measured by immunoassays, was highest in the IgGRF fraction, which bound most avidly to the IgG-Sepharose. When the oligosaccharides were released by hydrazinolysis, and analyzed by MALDI-TOF mass spectrometry and HPLC, in combination with sequential exoglycosidase treatment, all the IgG samples were found to contain a series of biantennary complex-type oligosaccharides. The incidence of galactose-free oligosaccharides was significantly higher in both IgGRFs and non-RF IgG from RA patients compared with IgG from healthy individuals. In all IgGRFs, the levels of sialylation and galactosylation were lower than those in non-RF IgG from RA patients; the sialylation of non-RF IgG was the same as that of IgG from healthy individuals. In addition, the decreases in galactosylation and sialylation of oligosaccharides in IgGRF correlated well with the increase in RF activity. These findings could contribute to our understanding of the mechanisms of IgG-IgG complex formation and the pathogenicity of these complexes in RA patients. PMID- 11011143 TI - Kinetic basis for the donor nucleotide-sugar specificity of beta1, 4-N acetylglucosaminyltransferase III. AB - The kinetic basis of the donor substrate specificity of beta1, 4-N acetylglucosaminyltransferase III (GnT-III) was investigated using a purified recombinant enzyme. The enzyme also transfers GalNAc and Glc moieties from their respective UDP-sugars to an acceptor at rates of 0.1-0.2% of that for GlcNAc, but Gal is not transferred at a detectable rate. Kinetic analyses revealed that these inefficient transfers, which are associated with the specificity of the enzyme, are due to the much lower V(max) values, whereas the K(m) values for UDP-GalNAc and UDP-Glc differ only slightly from that for UDP-GlcNAc. It was also found that various other nucleotide-Glc derivatives bind to the enzyme with comparable affinities to those of UDP-GlcNAc and UDP-Glc, although the derivatives do not serve as glycosyl donors. Thus, GnT-III does not appear to distinguish UDP-GlcNAc from other structurally similar nucleotide-sugars by specific binding in the ground state. These findings suggest that the specificity of GnT-III toward the nucleotide-sugar is determined during the catalytic process. This type of specificity may be efficient in preventing a possible mistransfer when other nucleotide-sugars are present in excess over the true donor. PMID- 11011145 TI - Monoclonal antibodies recognizing surface residues of the beta subunit of Escherichia coli F(1) ATPase: functional importance of the epitope residues. AB - Two monoclonal antibodies, beta 208 and beta 210, against the beta subunit of the F(1) ATPase from Escherichia coli reacted with an intact beta subunit and also a peptide corresponding to a portion of beta between residues 1 and 145. Mutations at Ala-1, Val-15, Glu-16, Phe-17, Leu-29, Gly-65, or Leu-66, and His-110 or Arg 111 for beta 210 and beta 208, respectively, caused decreased antibody binding to beta, suggesting that these residues form the epitopes and are thought to lie close together on the surface of the beta subunit. The topological locations of the corresponding residues in the atomic structure of the bovine beta subunit agree well with these expectations, except for Ala-1 and Leu-29. beta 210 binds to two beta strands including the epitope residues that are 50 residues apart, indicating that this antibody recognizes the tertiary structure of the N-terminal end region. Mutations in the epitope residues of beta 210 do not affect the F(1) ATPase activity, suggesting that surfaces of the two beta strands in the amino terminal end region are not functionally essential. To analyze the functional importance around His-110 recognized by beta 208 we introduced site specific mutations at residues His-110 and Ile-109. Ile-109 to Ala or Arg, and His-110 to Ala or Asp caused defective assembly of F(1). However, the His-110 to Arg mutation had no effect on molecular assembly, suggesting that Ile-109 and His 110, especially the positive charge of His-110 are essential for the assembly of F(1). The His-110 to Arg mutation caused a large decrease in F(1)-ATPase activity, suggesting that a subtle change in the topological arrangement of the positive charge of His-110 located on the surface of beta plays an important role in the catalytic mechanism of the F(1)-ATPase. PMID- 11011146 TI - Calcium-dependent and -independent hetero-oligomerization in the synaptotagmin family. AB - Synaptotagmins constitute a family of membrane proteins that are characterized by one transmembrane region and two C2 domains. Recent genetic and biochemical studies have indicated that oligomerization of synaptotagmin (Syt) I is important for expression of function during exocytosis of synaptic vesicles. However, little is known about hetero-oligomerization in the synaptotagmin family. In this study, we showed that the synaptotagmin family is a type I membrane protein (N(lumen)/C(cytoplasm)) by introducing an artificial N-glycosylation site at the N-terminal domain, and systematically examined all the possible combinations of hetero-oligomerization among synaptotagmin family proteins (Syts I-XI). We classified the synaptotagmin family into four distinct groups based on differences in Ca(2+)-dependent and -independent oligomerization activity. Group A Syts (III, V, VI, and X) form strong homo- and hetero-oligomers by disulfide bonds at an N-terminal cysteine motif irrespective of the presence of Ca(2+) [Fukuda, M., Kanno, E., and Mikoshiba, K. (1999) J. Biol. Chem. 274, 31421 31427]. Group B Syts (I, II, VIII, and XI) show moderate homo-oligomerization irrespective of the presence of Ca(2+). Group C synaptotagmins are characterized by weak Ca(2+)-dependent (Syts IX) or no homo-oligomerization activity (Syt IV). Syt VII (Group D) has unique Ca(2+)-dependent homo-oligomerization properties with EC(50) values of about 150 microM Ca(2+) [Fukuda, M., and Mikoshiba, K. (2000) J. Biol. Chem. 275, 28180-28185]. Syts IV, VIII, and XI did not show any apparent hetero-oligomerization activity, but some sets of synaptotagmin isoforms can hetero-oligomerize in a Ca(2+)-dependent and/or -independent manner. Our data suggest that Ca(2+)-dependent and -independent hetero-oligomerization of synaptotagmins may create a variety of Ca(2+)-sensors. PMID- 11011147 TI - Nitric oxide inactivates glyoxalase I in cooperation with glutathione. AB - We previously found that glyoxalase I (Glo I) is inactivated upon exposure of human endothelial cells to extracellular nitric oxide (NO), and this event correlates with an increase in its pI on two-dimensional gels. In this study, we demonstrate that NO can modulate Glo I activity in cooperation with cellular glutathione (GSH). Severe depletion of intracellular GSH prevents the inactivation of Glo I in response to NO, although such depletion enhances the inactivation of glyceraldehyde-3-phosphate dehydrogenase (G3PDH), a well-known enzyme susceptible to NO-induced oxidation. S-Nitrosoglutathione (GSNO), an adduct of GSH and NO, lowers the activity of purified human Glo I, while S nitrosocysteine (CysNO) inactivates the enzyme only in the presence of GSH. This indicates that a dysfunction in Glo I would require the formation of GSNO in situ. Competitive inhibitors of Glo I, S-(4-bromobenzyl)glutathione and its membrane-permeating form, completely abolish the NO action in vitro and inside cells, respectively. Taken together, these results reveal that Glo I can interact directly with GSNO, and that the interaction converts Glo I into an inactive form. Moreover, the data suggest that the substrate recognition site of Glo I might be involved in the interaction with GSNO. PMID- 11011148 TI - Synthesis and characterization of the spore proteins of Bacillus subtilis YdhD, YkuD, and YkvP, which carry a motif conserved among cell wall binding proteins. AB - We have previously reported that YaaH and YrbA are spore proteins of Bacillus subtilis that are required for spore resistance and/or germination and that they have a motif conserved among so-called cell wall binding proteins [Kodama et al. (1999) J. Bacteriol. 181, 4584-4591, Takamatsu et al. (1999) J. Bacteriol. 181, 4986-4994]. In this study, we analyzed the expression of ydhD, ykuD, and ykvP genes, which encode putative proteins containing the same motif. Transcription of ydhD was dependent on SigE, and the mRNA was detectable from 2 h after the cessation of logarithmic growth (T(2) of sporulation). ykuD was transcribed by SigK RNA polymerase from T(4) of sporulation. Both SigK and GerE were essential for ykvP expression, and this gene was transcribed from T(5) of sporulation. Inactivation of these genes by insertion of an erythromycin resistance gene did not affect vegetative growth, spore resistance to heat, chloroform, and lysozyme, or spore germination in the presence of L-alanine or in a mixture of L asparagine, D-glucose, D-fructose, and potassium chloride. The His tag fusions of YdhD, YkuD, and YkvP downstream of their natural promoter regions were introduced into a multicopy plasmid. These fusion proteins were produced during sporulation in B. subtilis transformants and were detected in mature spores, indicating that YdhD, YkuD, and YkvP are all proteins intrinsic to spores. Excessive YkuD and YkvP in the sporulating cells did not affect spore resistance or germination. The cells producing excessive YdhD also did not show impaired spore resistance, but their germination properties were changed: the spores revealed reduced response to L-alanine and some of them germinated even without germinants. Escherichia coli b-lactamase, whose signal sequence had been genetically replaced by the cell wall binding motif of YaaH, was produced in sporulating cells, and Western blot analysis indicated that the fused protein was assembled into spores. We speculate that the conserved motif functions as a kind of signal sequence involved in assembly of these proteins on forespores. PMID- 11011149 TI - Multiple actin-related proteins of Saccharomyces cerevisiae are present in the nucleus. AB - An increasing number of actin-related proteins (Arps), which share the basal structure with skeletal actin but possess distinct functions, have been found in a wide variety of organisms. Individual Arps of Saccharomyces cerevisiae were classified into Arps 1-10 based on the relatedness of their sequences and functions, where Arp1 is the most similar to actin, and Arp10 is the least similar. While Arps 1-3 and their orthologs in other organisms are localized exclusively in the cytoplasm, Arp4 (also known as Act3) is localized in the nucleus and is involved in transcriptional regulation. Here we examined the more divergent Arps for possible nuclear functions. We show that Arps 5-9 are localized in the nucleus, but Arp10 is not. The nuclear export signals identified in actin are well conserved in the cytoplasmic Arps, Arps 1-3, but less conserved in the nuclear Arps. Gel filtration chromatography experiments show that the nuclear Arps are larger than monomer in size and thus are present in multi protein complexes. Since nuclear protein complexes containing Arps are found to be responsible for histone acetylation and chromatin remodeling, it is suggested that most of the divergent Arps are involved in the !transcriptional regulation through chromatin modulation. PMID- 11011150 TI - Substrate recognition mechanism of prolyl aminopeptidase from Serratia marcescens. AB - Molecular cloning of the gene and the crystal structure of the prolyl aminopeptidase [EC 3.4.11.5] from Serratia marcescens have been studied by us [J. Biochem. 122, 601-605 (1997); ibid. 126, 559-565 (1999)]. Through these studies, Phe139, Tyr149, Glu204, and Arg136 were estimated to be concerned with substrate recognition. To elucidate the details of the mechanism for the substrate specificity, the site-directed mutagenesis method was applied. The F139A mutant showed an 80-fold decrease in catalytic efficiency (k(cat)/K(m)), but the Y149A mutant did not show a significant change in catalytic efficiency. The catalytic efficiency of the E204Q mutant was about 4% of that of the wild type. The peptidase activity of the mutant (R136A) was markedly decreased, however, arylamidase activity with Pyr-bNA was retained as in the wild-enzyme. From these results, it was clarified that the pyrrolidine ring and the amino group of proline at the S1 site were recognized by Phe139 and Glu204, respectively. P1' of a substrate was recognized by Arg136. On the other hand, the enzyme had two cysteine residues. Mutants C74A and C271A were inhibited by PCMB, but the double mutated enzyme (C74/271A) was resistant to it. PMID- 11011151 TI - Stereochemistry of the transamination reaction catalyzed by aminodeoxychorismate lyase from Escherichia coli: close relationship between fold type and stereochemistry. AB - Aminodeoxychorismate lyase is a pyridoxal 5'-phosphate-dependent enzyme that converts 4-aminodeoxychorismate to pyruvate and p-aminobenzoate, a precursor of folic acid in bacteria. The enzyme exhibits significant sequence similarity to two aminotransferases, D-amino acid aminotransferase and branched-chain L-amino acid aminotransferase. In the present study, we have found that aminodeoxychorismate lyase catalyzes the transamination between D-alanine and pyridoxal phosphate to produce pyruvate and pyridoxamine phosphate. L-Alanine and other D- and L-amino acids tested were inert as substrates of transamination. The pro-R hydrogen of C4' of pyridoxamine phosphate was stereospecifically abstracted during the reverse half transamination from pyridoxamine phosphate to pyruvate. Aminodeoxychorismate lyase is identical to D-amino acid aminotransferase and branched-chain L-amino acid aminotransferase in the stereospecificity of the hydrogen abstraction, and differs from all other pyridoxal enzymes that catalyze pro-S hydrogen transfer. Aminodeoxychorismate lyase is the first example of a lyase that catalyzes pro-R-specific hydrogen abstraction. The result is consistent with recent X-ray crystallographic findings showing that the topological relationships between the cofactor and the catalytic residue for hydrogen abstraction are conserved among aminodeoxychorismate lyase, D-amino acid aminotransferase and branched-chain L-amino acid aminotransferase [Nakai, T., Mizutani, H., Miyahara, I., Hirotsu, K., Takeda, S., Jhee, K.-H., Yoshimura, T., and Esaki, N. (2000) J. Biochem. 128, 29-38]. PMID- 11011152 TI - Solution structure of myosin-ADP-MgFn ternary complex by fluorescent probes and small-angle synchrotron X-ray scattering. AB - In the presence of excess amounts of fluorine, a physiological divalent cation, magnesium (Mg(2+)), forms a novel phosphate analogue, magnesium fluoride (MgFn). Park et al. [Biochim. Biophys. Acta 1430, 127-140 (1999)] previously demonstrated that MgADP. MgFn forms a complex with myosin subfragment-1 (S-1), and the S 1.ADP. MgFn ternary complex mimics a transient state in the activity cycle of ATPase. In the present study, localized conformations in the regions of highly reactive cysteine and lysine residues, Cys 707 (SH1), Cys 697 (SH2), and Lys 83 (RLR), which change their conformations markedly during ATP hydrolysis, were studied using fluorescent probes and chemical modification. The global shape of the complex was also studied using small angle X-ray solution scattering and compared it with other previously reported myosin.ADP. fluorometal ternary complexes. The results suggest that the overall conformation and localized functional regions of the complex are quite similar to those in the presence of ATP, indicating that the complex mimics the M(**).ADP.P(i) steady state. PMID- 11011153 TI - Conformational changes in the unique loops bordering the ATP binding cleft of skeletal muscle myosin mediate energy transduction. AB - Myosin has three highly-conserved, unique loops [B (320-327), M (677-689), and N (127-136)] at the entrance of the ATP binding cleft, and we previously showed that the effects of actin are mediated by a conformational change in loop M [Maruta and Homma (1998) J. Biochem. 124, 528-533]. In the present study, loops M and N were photolabeled respectively with fluorescent probes Mant-8-N(3)-ADP and Mant-2-N(3)-ADP in order to study conformational changes in the loops related to energy transduction. The effect of actin on the conformation of loop N was examined by analyzing fluorescence polarization and acrylamide quenching; the results were then compared with those previously reported for loop M. In contrast to loop M, the fluorescence polarization and the value of K(sv) of the Mant groups crosslinked to loop N were slightly affected by actin binding. To study conformational changes in loops M and N during the ATPase cycle, FRET was analyzed using TNP-ADP.BeFn and TNP-ADP. AlF(4)(-) as FRET acceptors of Mant fluorescence. The resultant estimated distances between loop M and the active site differed for the Mant-S1.TNP-ADP.BeFn and Mant-S1.TNP-ADP.AlF(4)(-) complexes, whereas the distances between loop N and the active site differed slightly. These findings indicate that the conformation of loop M changes during the ATPase cycle, suggesting that Loop M acts as a signal transducer mediating communication between the ATP- and actin-binding sites. Loop N, by contrast, is not significantly flexible. PMID- 11011154 TI - Fibrinogen binds to integrin alpha(5)beta(1) via the carboxyl-terminal RGD site of the Aalpha-chain. AB - Fibrinogen interactions with vascular endothelial cells are implicated in various physiological and pathophysiological events, including angiogenesis and wound healing. We have shown previously that integrin alpha(5)beta(1) is a fibrinogen receptor on endothelial cells [Suehiro, K., Gailit, J., and Plow, E.F. (1997) J. Biol. Chem. 272, 5360-5366]. In the present study, we have characterized fibrinogen interactions with purified alpha(5)beta(1) and have identified the recognition sequence in fibrinogen for alpha(5)beta(1). The binding of fibrinogen to immobilized alpha(5)beta(1) was selectively supported by Mn(2+). Fibrinogen bound to purified alpha(5)beta(1) in a time-dependent, specific, and saturable manner in the presence of Mn(2+), and the binding was blocked completely by Arg Gly-Asp (RGD)-containing peptides and by anti-alpha(5) and anti-alpha(5)beta(1) monoclonal antibodies. A monoclonal antibody directed to the C-terminal RGD sequence at Aalpha572-574 significantly inhibited the binding of fibrinogen to alpha(5)beta(1), whereas monoclonal antibodies directed to either the N-terminal RGD sequence at Aalpha95-97 or the C-terminus of the gamma-chain did not. Furthermore, substituting RGE for RGD at position Aalpha95-97 in recombinant fibrinogen had a minimal effect on binding, whereas substituting RGE for RGD at position Aalpha572-574 decreased binding by 90%. These results demonstrate that the C-terminal RGD sequence at Aalpha572-574 is required for the interaction of fibrinogen with alpha(5)beta(1). PMID- 11011156 TI - [The association of French-speaking dermatologists]. PMID- 11011155 TI - A half-type ABC transporter TAPL is highly conserved between rodent and man, and the human gene is not responsive to interferon-gamma in contrast to TAP1 and TAP2. AB - TAPL is a half-type ABC transporter with sequence similarity to TAP1 and TAP2 that is transcribed in various rat tissues [Yamaguchi, Y., Kasano, M., Terada, T., Sato, R., and Maeda, M. (1999) FEBS Lett. 457, 231-236]. Primary structures of the human and mouse orthologous counterparts were deduced from cDNAs cloned by means of polymerase chain reaction, and they were compared with that of the rat. The mammalian TAPLs (rat, mouse, and human) are highly conserved, since about 95% of the amino acid residues are identical between rodents and man. Phylogenetic analysis demonstrated that the evolutional rate of TAPL is much slower than those of TAP1 and TAP2, although TAPL could have diverged from an ancestor of TAP1 or that of TAP1 and TAP2. The TAPL-GFP fusion protein transiently expressed in Cos-1 cells was co-localized with PDI, suggesting that TAPL is inserted into endoplasmic reticulum membrane. The conservation of the peptide-binding motifs of TAP proteins in TAPL raises the possibility that the TAPL might be a peptide transporter. The gene for human TAPL is assigned to chromosome 12q24.31-q24.32, while those for TAP1 and TAP2 are located at the MHC locus of chromosome 6p21.!3. Furthermore, the transcription of TAPL gene is not responsive to interferon gamma, in contrast to TAP1 and TAP2. These results indicate that the gene regulation of TAPL is different from those of TAP1 and TAP2. PMID- 11011157 TI - [Should immunosuppressors be used for the treatment of atopic dermatitis?]. PMID- 11011158 TI - [Bioavailability evaluation of dermocorticoids using differential infrared thermography]. AB - INTRODUCTION: Differential infrared thermography, proposed in this paper, is a technique based on direct observations of infrared radiations emitted by the skin. The evolution of cutaneous temperature caused by the application of dermocorticoids on healthy skin demonstrates their pharmaco-dynamic properties. MATERIALS AND METHODS: The cutaneous thermal image was recorded in real time. Image processing using subtraction function readily provided differential infrared thermographic analysis of the effects. Four activity classes of dermocorticoids had been applied on healthy skin. A test immediately carried out after dermocorticoid application on skin without occlusion and the classical skin blanching-test have been performed. RESULTS: Temperature differences between the dermocorticoids were detected within the first three hours after the application on the skin without occlusion. The dermocorticoid class II cream formulation under study induced a decrease in temperature more pronounced than the others dermocorticoids. The skin-blanching effect was more noticeable for dermocorticoids class I and II and it was not detected for class IV. DISCUSSION: Subtraction thermograms provide a means of differential imaging. Evolutive temperature differences subsequent to unique application without dermocorticoid occlusion are evidenced during a short duration (the first three hours). This may correspond to efficacy differences while classical tests of vasoconstriction analyse the cutaneous blanching induced after the 6th hour. Concerning the skin blanching effect, results of this first investigation are not sufficient for a precise qualitative and quantitative interpretation. The main interest of differential infrared thermography is to be quantitative, without contact, continuous in real time. Differential infrared thermography is more sensitive than classical thermography. It allowed an objective evolution survey for dermocorticoids. PMID- 11011159 TI - [Herpetiform ulceration: 5 cases]. AB - BACKGROUND: Mouth ulcers are frequently observed, mostly caused by trauma and aphthae. We report five cases of herpetiform ulceration, a rare clinical form of aphthae often mistaken for herpetic stomatitis because of the similar clinical presentation. CASE REPORTS: All patients were men with late age at onset (44.8 years). All had numerous tiny very painful ulcers covering all parts of the oral cavity. Consequences were sometimes severe (weight loss and general weakness in two patients, requiring hospitalization in one). Diverse clinical courses were observed (from no recurrence to monthly recurrence). Systemic steroids resulted in dramatic improvement in two patients, colchicine prevented recurrences in three of them. DISCUSSION: These observations illustrate well this rare disease: herpetiform ulceration. We found oral corticosteroids provide effective cure and observed the preventive action of colchicine. Both findings should be confirmed in larger series. PMID- 11011160 TI - [Gonorrhea in sexually abused young girls in Lome (Togo)]. AB - INTRODUCTION: Sexually transmitted diseases due to a sexual abuse cause are rarely documented in black Africa. The purpose of this study was to demonstrate the existence of sexual abuse in young girls with gonorrhea observed in Lome (Togo). PATIENTS AND METHODS: A cross-sectional study was conducted to document cases of sexually transmitted diseases diagnosed in young children (0 to 11 years of age) seen at the dermato-venereology unit of the Lome teaching hospital over a 20 month period. Syphilis serology (TPHA-VDRL) and HIV serology were carried out for all children with sexually transmitted disease and repeated after two weeks for TPHA-VDRL, and three months for HIV serology in children who had been sexually abused. RESULTS: During this period, 13 of 33 cases of sexually transmitted diseases diagnosed in young children were gonorrhea (mean age 7.2 +/- 2.7 years). It was due to sexual abuse in 12 cases (all in young girls). The abuser was a domestic employee in the child's home (n =3), a member of the child's family (n =7), an educator (n =1), a neighbor (n =1). Mean age of the alleged authors of sexual abuse was 25.7 +/- 5.5 years. Syphilis serology was negative, but one case of HIV infection in a 10-year-old girl was observed with identification of the contaminator. CONCLUSION: The results of this study confirm that sexual abuse in children is not an uncommon occurrence in black Africa and that it often leads to gonorrhea. The classic consequences of such abuse are aggravated by the high prevalence of HIV infection observed in the majority of the countries in black Africa. PMID- 11011161 TI - [Eosinophilic ulceration of the tongue]. AB - BACKGROUND: Eosinophilic ulceration of the tongue is a classic condition little reported in the literature. We describe a case in a patient taking nicorandil. CASE REPORT: An 84-year-old patient consulted for an ulceration of the tongue which had progressed for 5 months. The ulcer was very painful and the patient had lost 2.5 kg. The patient had been taking nicorandil at the dose of 10 mg/d for a year. Biopsies of the tongue ulcer confirmed the diagnosis of eosinophilic ulceration. Immunohistochemistry determined predominantly T-cell infiltration. Search for Epstein-Barr virus was negative. The ulcer completely regressed within one month without withdrawal of nicorandil. DISCUSSION: This was a typical case of eosinophilic ulceration of the tongue, both from the point of view of the clinical expression and the histological findings. A nicorandil-induced ulcer was ruled out on clinical (low daily dose, delay to onset) and histological arguments as well as the spontaneous regression without drug withdrawal. Nicorandil may have played a role in the abnormally long duration of the ulceration. PMID- 11011162 TI - [Localized bullous pemphigoid induced by thermal burn]. AB - BACKGROUND: Induced bullous pemphigoid is known to be triggered by drugs and some physical agents. Six cases of bullous pemphigoid induced by thermal burns have been reported since 1991. CASE REPORT: We observed a bullous eruption around thermal burns of the left leg in a 89 year-old woman. Bullous pemphigoid was diagnosed by histological and immunohistochemical findings with a sub-epidermal blister and linear IgG and C3 deposition at the dermal-epidermal junction. Indirect immunofluorescence was negative. The eruption rapidly resolved with topical steroids. DISCUSSION: We describe a bullous pemphigoid induced by a thermal burn. This case is original because the eruption was localized only around the site of the burn and healed with topical steroids. In the literature, there are 6 other cases reported but only one localized. We discuss the hypothesis of asymptomatic bullous pemphigoid exacerbated by presentation of bullous pemphigoid antigen by thermal burns or self immunization against antigens altered by burns. PMID- 11011163 TI - [Granuloma annulare of the photoexposed areas in two liver transplant recipients]. AB - BACKGROUND: We report two cases of generalized granuloma annulare occurring in photoexposed areas in two liver transplant recipients. CASE REPORTS: Case 1 was a 65-year-old man who had undergone liver transplantation in 1992. He was given immunosuppressive and antihypertensive therapy. Within 18 months of transplantation, he developed a confluent rash with maculae on sun-exposed areas (neck opening, nape, arms) and sparing the undershirt area. Clinical examination was normal. Skin biopsy revealed a palissade infiltrate located in the middle and upper derma, suggestive of granuloma annulare, with elastophagocytosis patterns (orcein stain and ultrastructure study). Photoexposure granuloma was diagnosed. Case 2 was a 59-year-old man who had undergone liver tranplantation in 1994. He was given immunosuppressive and antihypertensive therapy and developed within 4 months a dark rash on the neck opening and nape, sparing photoprotected areas. Histopathology revealed granuloma annulare. Elastophagocytosis was disclosed by orcein stain and the ultrastructure study. DISCUSSION: This clinical presentation of granuloma annulare in two liver transplant recipients is unusual. We discuss the clinical and histopathological patterns observed in our two cases and the relationships between granuloma annulare and immunosuppression. PMID- 11011164 TI - [Microsporum canis mycetoma of the scalp]. AB - BACKGROUND: Mycetoma is a chronic subcutaneous tumefaction with presence of grains or granules. Etiological agents include bacteria or filamentous fungi. Mycetoma due to dermatophytes is uncommon, mainly occurring in Africa. To our knowledge, no case has been reported in the West Indies. Only two observations of Micosporum canis mycetoma in humans have been reported in the literature. We report a third case of mycetoma of the scalp caused by this fungus. CASE REPORT: A 22-year-old woman from Martinique, French West Indies, presented with an indolent tumefaction of the scalp evolving over five years. She had mental retardation due to congenital adrenal hyperplasia with 21-hydroxylase deficiency. The lesion was extracted surgically. Pathology and mycology examinations showed features of Microsporum canis mycetoma. Two months later, the scalp lesion recurred and the patient was treated with griseofulvin after surgical extraction. DISCUSSION: Mycetoma due to dermatophytes is very uncommon, mainly observed on the scalp and nape of the neck. A history of a skin lesion is frequent, leading to transcutaneous penetration of the fungus and mycetoma formation. Several dermatophyte species have been identified as causal agents (Microsporum ferrugineum, Trichophyton rubrum, Trichophyton verrucosum, Trichophyton mentagrophytes, Microsporum audouinii, Microsporum langeronii). Microsporum canis is rarely demonstrated in humans: two cases in children in Africa and Australia. Our observation was similar to the two cases in the literature: indolent and mobile tumefaction of the scalp, in a child or young adult, suggestive of lipoma or epidermal cyst, with excision leading to diagnosis. Association with tinea capitis and skin or nail involvement can also be observed. PMID- 11011165 TI - [Lymphoma presenting as Sister Mary-Joseph's nodule]. AB - BACKGROUND: Cutaneous metastasis of the umbilicus is generally associated with intra-abdominal adenocarcinoma, rarely with a malignant hemopathy. CASE REPORT: We report a case of lymphoma presenting as Sister Mary-Joseph's nodule, in a 79 year-old man. Histology evidenced small-cell lymphoma. DISCUSSION: Umbilical metastasis is uncommon, most of these lesions arising from gastric adenocarcinoma. Only two cases of lymphoma, associated with an umbilical metastasis have been reported. This case opens the discussion on the fact that the umbilicus is generally not a site of metastatic spread of lymphomas. PMID- 11011166 TI - [Erythema annulare centrifugum and relapsing polychondritis]. AB - BACKGROUND: Relapsing polychondritis is a rare systemic disease. Skin involvement occurs in 20 to 50 % of cases. Cutaneous signs are most often related to a leukocytoclastic vasculitis. Association of relapsing polychondritis with neutrophilic dermatosis have also been reported. We report the first case of an erythema annulare centrifugum-like dermatosis associated with relapsing polychondritis, with a two years delay between both conditions. CASE REPORT: A 74 year-old man was seen for papulo-erythematous centrifugal annular lesions that appeared 18 months earlier in a context of bad general conditions. Biological tests were normal. Several skin biopsies were performed, showing at the beginning features of drug reaction and then of lupus-lichen. Treatment with hydroxychloroquine, topical corticosteroids, dapsone and thalidomide were unsuccessful. In the following months, the patient developed fever and relapsing bronchitis. Suddenly, a chondritis of the ears appeared, leading to the diagnosis of relapsing polychondritis. All the cutaneous, chondritic and respiratory signs disappeared with oral steroid therapy. Two years after the diagnosis of relapsing polychondritis the patient developed refractory anemia. DISCUSSION: Cutaneous signs of relapsing polychondritis are frequent and may occur several months or years before the chondritis. They are polymorphous, but to the best of our knowledge, a clinical aspect of erythema annulare centrifugum has never been described. Our observation recalls the sometimes long delay between the cutaneous and the chondritic signs of relapsing polychondritis and the high frequency of dysmyelopoiesis in relapsing polychondritis with cutaneous involvement. PMID- 11011167 TI - [Cutaneous manifestations of Yersinia enterocolitica infection]. AB - BACKGROUND: Cutaneous manifestations occurring in infections due to Yersinia enterocolitica are usually erythema nodosum, erythema multiforme or cutaneous vasculitis. The association between Yersinia infection and Sweet's syndrome is rare. We describe such a case contributing to the discussion on this association. CASE-REPORT: A 29-year-old woman had a papulo-pustular eruption with fever associated with arthralgia. The results of the infectious laboratory investigations were negative but Yersinia enterocolitica type 0.9 was isolated from a stool culture. The serologic diagnosis of Yersinia enterocolitica using serum agglutinins was negative. The diagnosis of Sweet's syndrome was made on a skin biopsy specimen. Search for hematology disease or underlying neoplasia was negative. The clinical course was rapidly favorable with antibiotic treatment (ciprofloxacin). DISCUSSION: The diagnosis of Yersinia enterocolitica infection is difficult. Microbiologic diagnosis of Yersinia infection is best achieved by isolation of the bacterium from a clinical specimen of involved tissue. The agglutination test is not highly specific or sensitive. Immunoblotting appears to be more sensitive. PMID- 11011168 TI - [Comments on the 'Case for diagnosis' reported by Vieira-mota et al]. PMID- 11011170 TI - [Wasp stings with localized eruption]. PMID- 11011172 TI - [Hypocromatic spots on pigmented skin]. PMID- 11011171 TI - [Familial disseminated amylosis with cutaneous and cardiac predominance by apolipoprotein A1 mutation]. PMID- 11011173 TI - [Ecchymotic aeras to biopsy]. PMID- 11011174 TI - [Recommendations of the French Society of Photodermatology for systemic PUVA therapy in psoriasis vulgaris. French Society of Photodermatology]. PMID- 11011176 TI - [Cell therapy: perspectives for curing skin wounds?]. PMID- 11011175 TI - [Management of pain]. PMID- 11011178 TI - [New nail and hair side-effects of cyclosporin]. PMID- 11011177 TI - [Erythematous pemphigus (seborrheic pemphigus)]. PMID- 11011179 TI - [Challenge test for toxic skin reactions to drugs]. PMID- 11011180 TI - [DermIS, University of Erlangen at Nuremberg]. PMID- 11011182 TI - [Guichard Joseph Duverney (1648-1730), first French otologist in the 17th century]. AB - PURPOSE OF STUDY: Study and understand the role of Guichard Joseph Duverney in the history of French otology. METHOD: Study of the different works and articles published about him, linked with reading his principal writings. RESULTS: Duverney published one of the first complete works on otology titled Traite de l'organe de l'ouie (Treatise of the organ of hearing) in 1683. Following a first part containing numerous new notions of anatomy, Duverney developed in the second part a theory of hearing very similar to that of von Helmholtz. In the third part, he finally cover several pathologies of the ear without great innovation. Translated into many languages, this book became the book of reference for all the otologists of the 17th and 18th centuries. CONCLUSION: Duverney is rightly considered to be the first French otologist of the 17th century and also, as certainly one of the first European otologists. PMID- 11011183 TI - [Maurice Sourdille and the operating microscope]. AB - The authorship of the operating microscope appears to be shared by Nylen, who had the idea of using a microscope for ear surgery and Gullstrand, who invented the slit-lamp and who was probably the counsellor of Holmgren in his work on operative field magnification. Holmgren was the first to use an operating binocular magnifying glass and Sourdille created the first binocular magnifying glass designed for microsurgery. PMID- 11011184 TI - [Tympanoplasty and Sourdille]. AB - In his article published earlier in 1929, Maurice Sourdille proposed two innovations in middle ear surgery: the tympanomeatal flap and surgery of chronic otitis. Those events were not only the starting point of a new era of modern tympanoplasty but also led to the way to the development of middle ear exploration with purposeful resection of the tympanic framework for accessing to the windows and for sealing a perforated tympanic membrane. PMID- 11011185 TI - [A century in the history of chronic ear surgery]. AB - Although some advocated surgery of the tympanic membrane and stapes as early as early as the end of the nineteenth century, it took a long time for widespread development. Mastoid surgery had just been born and was to be greatly improved over the next century. Preservation of the tympano-ossicular its later reconstruction gradually came into use with the creation of the tympanomeatal flap and the recent development of middle ear surgery, followed by posterior tympanotomy and myringoplasty, thus allowing closed tympanoplasty. Likewise, the ventilation tube, forgotten for more than half a century, has found an important place in the treatment of chronic otitis with a closed ear drum. PMID- 11011186 TI - [A century of ostospongiose or the "century" of ostospongiosis?]. PMID- 11011187 TI - [A hundred years of vertigo]. PMID- 11011188 TI - [A century of ENT cancerology]. PMID- 11011217 TI - Potential influence of lipids in diabetic nephropathy: insights from experimental data and clinical studies. AB - Diabetic nephropathy is associated with an altered lipid profile characterized by elevated triglyceride rich lipoproteins, present even in the earlier stages of the renal disease. Although many experimental studies have demonstrated a significant deleterious role for hyperlipidemia in both the initiation and progression of renal injury, data remain more conflicting in humans. A few prospective studies, mostly in type 2 diabetes, have suggested an independent role for serum cholesterol level in the subsequent development of incipient or overt diabetic nephropathy. Furthermore, studies have reported in both types of diabetes an independent deleterious influence of serum total cholesterol on the decline in renal function and/or progression of albuminuria. However, the majority of these studies were post hoc analyses of previously controlled therapeutic trials with several observational studies not confirming these findings. It remains controversial whether apolipoprotein E gene polymorphism is an important factor in the development of diabetic nephropathy. Most of the interventional studies with lipid-lowering therapy in diabetic nephropathy have used HMG CoA reductase inhibitors and have been inconclusive. This may be due to a too short follow-up or insufficient number of patients. Further larger prospective studies are therefore required to better ascertain the role of lipids in the progression of diabetic nephropathy. PMID- 11011218 TI - Post-prandial hyperglycemia. post-prandial hyperglycemia and diabetes. AB - Post-prandial hyperglycemia (PPHG) is an independent risk factor for the development of macrovascular complications. It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose. Many factors affect the post-prandial blood glucose excursion. The glycemic index of the meal depends on the nature of the ingested food and starch composition. Gastric emptying is influenced by various factors including gut hormones such as GIP and GLP1, which potentiate insulin secretion, especially in its acute first phase, now referred to as an incretin effect. They also modulate glucagon secretion. Post-prandial hyperglycemia is limited by uptake of glucose by the liver and by inhibition of endogenous glucose production in this organ. In healthy controls, hepatic glucose production is halved after a meal, whereas in glucose-intolerant individuals and type 2 diabetics this inhibition is impaired (20-30% versus 50%). The persistence of endogenous glucose production during the post-prandial phase appears to be the main culprit in the PPHG. This reduced decrease in endogenous glucose in glucose intolerant and type 2 diabetic patients depends not only on the first acute phase of insulin secretion, but above all on the non-suppressed glucagon level during the post-prandial phase. Glucagon levels fall in healthy control subjects during the post-prandial phase. Although peripheral glucose uptake by insulin-dependent tissues is altered in type 2 diabetic patients, it does not appear to be the major cause of the PPHG as there are patients with insulin resistance but without post-prandial hyperglycemia. PMID- 11011219 TI - Contribution of total and intact proinsulins to hyperinsulinism in subjects with obesity, impaired glucose tolerance or type 2 diabetes. AB - The recent development of specific radioimmunoassays of insulin (Ins) and proinsulin (PI) led some authors to question the classical data on insulinosecretion in patients with abnormal glucose tolerance. The aim of this work was to determine the participation of intact proinsulin (iPI) and its split fragments to total insulinsecretion in obese subjects and in various stages of glucose intolerance determined by an oral glucose load according to the WHO recommendations. Five groups were constituted: non obese controls (C), obese subjects with normal glucose tolerance (O), non obese subjects with impaired glucose tolerance (I), obese subjects with impaired glucose tolerance (OI) and diabetic patients (D). The basal level of total Proinsulin (tPI) of D and OI was significantly higher than that of C, but the tPI/Ins ratio did not differ between the five groups. After glucose load, this ratio tended to be higher in D, but not significantly. No statistical difference between groups was observed for the iPI/Ins ratio. These results indicate that the determination of tPI is at least as informative or more than that of iPI. Furthermore, the proportionally constant participation of PI to insulin secretion observed in various stages of glucose intolerance suggests that the results obtained in the past with non specific insulin radioimmunoassays remain valid. PMID- 11011220 TI - The DECODE study. Diabetes epidemiology: collaborative analysis of diagnostic criteria in Europe. AB - The current criteria for the diagnosis of diabetes used in France are now based on those published by the American Diabetes Association in 1997: fasting plasma glucose >/= 7.0 mmol/l (126 mg/dl) (previously >/= 7.8 mmol/l (140 mg/dl)), 2 hour glucose >/= 11.1 mmol/l (200 mg/dl) following a 75g oral glucose tolerance test. However, while the American Diabetes Association recommended that the post charge test not be used, both the World Health Organisation and the French Language Association for the study of Diabetes and Metabolic diseases (ALFEDIAM) retained this test. The DECODE (Diabetes Epidemiology: Collaborative analysis of Diagnostic criteria in Europe) study analysed the effect of these changes on the prevalence of diabetes, and whether the changes were justified by the mortality in the various glycaemic groups, using epidemiological data on close to 30,000 subjects from twenty European epidemiological studies. The prevalence of diabetes, using fasting rather than the 2-hour glucose concentrations (as had previously been recommended for epidemiological studies) resulted in changes in the prevalence of diabetes, an increase or a decrease, depending on the population studied. The fasting criteria tended to diagnose younger and more obese subjects than the 2-hour criteria. The subjects who would now be diabetic with the new fasting diagnostic criteria suffered a high mortality, similar to that of other diabetic subjects, thus the new criteria can be justified. However, the diabetic subjects who only have a post-charge diabetic hyperglycaemia (>/= 11.1 mmol/l (200 mg/dl)), are now even less likely to be screened as diabetic, despite the fact that they have a risk of premature death of the same order as other diabetic subjects. PMID- 11011221 TI - What do the needles, syringes, lancets and reagent strips of diabetic patients become in the absence of a common attitude? About 1070 questionnaires in diabetic clinics. AB - The aim was to investigate the fate of injection and monitoring material after its use by diabetic patients in different countries (France, Belgium, Luxemburg, Switzeland and Tunisia). Some suitable containers are available for disposal but little is known about the attitudes of patients and physicians to them. 1 070 questionnaires were completed by patients (age: 50 +/- 18 years; diabetes duration: 15 +/- 11 years; 2.8 +/- 1.1 injections per day) visiting 109 doctors. Injections were done at home (72.6%), or both at home and at work (26.6%). At home: needles, syringes, lancets and reagent strips were thrown directly into the bin in 46.9%, 49.9%, 52.2% and 67.6% of cases, respectively; and in a closed plastic bottle in 29. 6%, 28.5%, 28.9% and 19.9% of cases, respectively. Specific containers were used in 8.6% and 6.3% of cases for needles and syringes, respectively. 62% of the bottles and containers were thrown directly into the bin, whereas 15.5% were returned to a pharmacy (4.5% taken to hospitals, 2.9% were burned). At work: 63% of the patients brought their needles and syringes home for disposal, 6.9% kept suitable containers at work and 30% threw their materials directly into local bins. We conclude that awareness should be increased and the organization of the collection of used material improved. PMID- 11011222 TI - Case complexity and clinical outcome in diabetes mellitus. A prospective study using the INTERMED. AB - The aim of this study was to assess a population of patients with diabetes mellitus by means of the INTERMED, a classification system for case complexity integrating biological, psychosocial and health care related aspects of disease. The main hypothesis was that the INTERMED would identify distinct clusters of patients with different degrees of case complexity and different clinical outcomes. Patients (n=61) referred to a tertiary reference care centre were evaluated with the INTERMED and followed 9 months for HbA1c values and 6 months for health care utilisation. Cluster analysis revealed two clusters: cluster 1 (62%) consisting of complex patients with high INTERMED scores and cluster 2 (38%) consisting of less complex patients with lower INTERMED. Cluster 1 patients showed significantly higher HbA1c values and a tendency for increased health care utilisation. Total INTERMED scores were significantly related to HbA1c and explained 21% of its variance. In conclusion, different clusters of patients with different degrees of case complexity were identified by the INTERMED, allowing the detection of highly complex patients at risk for poor diabetes control. The INTERMED therefore provides an objective basis for clinical and scientific progress in diabetes mellitus. Ongoing intervention studies will have to confirm these preliminary data and to evaluate if management strategies based on the INTERMED profiles will improve outcomes. PMID- 11011224 TI - [Survival analysis of diabetic patients on dialysis in Guadeloupe]. AB - BACKGROUND: Both diabetes and hypertension, two conditions that can lead to renal failure, have a high prevalence in Guadeloupe. OBJECTIVE: To determine the clinical and epidemiological features of diabetic patients on end stage renal failure and to evaluate their survival. PATIENTS AND METHOD: Data of the Guadeloupe Kidney registry were analysed for patients who began chronic dialysis during 1978-1997. Follow up information on survival status was obtained up to January 26(th) 1999. Cox proportional hazard analysis was used to determine the relative risk (RR) of death between levels of independant variables. RESULTS: There were 784 dialysis patients of whom 174 (22%) were diabetics. Among the latter, there were 97 women (55,7%), mean age at the start of dialysis was 60.6 years (range 26-83) and arterial hypertension was present before the start of dialysis in 67% of them. Median survival MS (95%CI) was significantly lower in diabetics 42 months (31-52) than in non diabetics 83 months (70-96), p<10(-4). In diabetics, the cumulative probability of survival was 83% (1 year) and 39% (5 years) and the RR of death (95% CI) were 1.90 (1.10-3.22) and 3.43 (2.00-5.87) for diabetics admitted in dialysis in age-class 55-64 years and 65-83 years, respectively, when that for age-class 54 years was set at 1. 00. The RR for diabetics was 1.67 (1.33-2.10) relative to non diabetics. CONCLUSION: Prospective studies are warranted to describe the role of comorbid conditions in diabetic patients survival. Prevention of degenerative complications should be a priority in this population. PMID- 11011223 TI - Possible activation of auto-immune thyroiditis from continuous subcutaneous infusion of genapol-containing insulin. AB - A case of a type 1 diabetic woman with auto-immune thyroiditis is reported, in whom repeated exposure to insulin containing Genapol(R) (polyethylen polypropylenglycol) over 3 years reproducibly parallels with an increase of serum TSH (thyroid-stimulating hormone) above the normal limit. Previously, adverse effects of Genapol(R) insulin have been related to its intraperitoneal application, and thought to be restricted to anti-insulin-immunity; activating effects on thyroid auto-immunity have been repeatedly disputed. We suggest that Genapol(R) insulin should be replaced by other insulin preparations with a better safety record. PMID- 11011225 TI - Telling about diabetes canhelp disease acceptance in indulin dependent diabetes. A clinical experience . AB - Type 1 diabetes (IDDM) is an original, chronical illness which concerns the physician as well as the patient, who is strongly implicated in the self management of his treatment. In order to help them to cope with their illness, 31 IDDM patients patients usually attending our diabetology unit were submitted to an open questionaire. We considered that IDDM may be a psycho-somatic disease, and that psychological factors may interfere with glycemic balance. The purpose was 1) to explore the patient's relationship to himself as a diabetic subject, while listening to his "story"; 2) if telling his story is able to reduce his burden or if there is a risk of setting down a psychic level. In fact, it allowed a better self-knowing and understanding, so that the patient could get back some power against illness; it appears a useful tool for a better dialog between the physician and the patient. PMID- 11011226 TI - [Is diabetes mellitus a sufficient condition to suspect hemochromatosis?]. AB - Genetic Hemochromatosis (GH) is a highly prevalent autosomal recessive disorder, which outcome has been dramatically improved by early phlebotomy. Attempts to screen for the disease, using biological and genetic approaches, are currently under evaluation. Diabetes mellitus often complicates GH. However, as it occurs late in the course of the disease, in most cases when cirrhosis is already present, its usefulness for the screening of GH seems reduced. Diabetes mellitus, when isolated, appears also as a poor predictor of hemochromatosis. Indeed, the risk of being carrier of the disease is not increased in diabetic patients compared with non diabetic sujects. This risk is however highly enhanced by the co-existence of cirrhosis. Thus, in the face of a newly diagnosed diabetes mellitus, the search for hemochromatosis must be performed only when it associates with cirrhosis or with other evocative clinical conditions. PMID- 11011228 TI - [Treatment of type 2 diabetes]. PMID- 11011227 TI - [Diabetic retinopathy in Dakar and review of African literature: epidemiologic elements]. AB - Literature review shows that in the African Sub-Sahara, prevalence of diabetic retinopathy is between 15 and 52%, the main age being the fifties (between 45.5 and 53.25 years in our cases); after 5 years of diabetes evolution (8.45 +/- 6.93 years in our cases) men being two to three fold more affected than women; non insulin dependent diabetes representing 77% and insulin dependent diabetes 23%. The type of retinopathy is usually mixed, both edematous and ischemic (88% being non proliferative). Retinopathy concerned mainly non obese, non insulin dependent diabetics in our cases. Risk factors are poor glycemic control and arterial hypertension. Diabetes duration was the sole and most evident risk factor. Diabetes duration was 5.28 +/- 5.28 years when retina was normal and reached 8.58 +/- 5.33 years for Stage II and 12.8 +/- 6.31 years for retinopathy Stage III and 16.75 +/- 7.25 years for retinopathy Stage IV. These factors, delayed diagnosis and diabetes progression in Africa justify improvement of diabetes care by multidisciplinary team to prevent blindness. PMID- 11011229 TI - [Introduction]. PMID- 11011230 TI - [General method]. PMID- 11011231 TI - [Strategy for documentary research]. PMID- 11011232 TI - [Strategies for care of the type 2 diabetic patient excluding care of complications. Recommendations of ANAES (National Agency for Health Accreditation and Evaluation March 2000]. PMID- 11011233 TI - [Health care in non-insulin dependent diabetics in lower Normandy]. AB - To contribute to better quality health care for patients with type 2 diabetes mellitus living in the French Department Basse-Normandie, the Regional Union of General Practitioners (URML) and the Regional Union of the Nation Health Insurance (URCAM) studied ambulatory practices in this population in 1999. Eight hundred diabetics whose health care reimbursements suggested they were not followed by a specialized center were studied: 400 received a mailed questionnaire and for the 400 others, another questionnaire was filed out by the primary care physician. The physicians' responses demonstrated that they do not feel certain recommended complementary investigations are always necessary for following diabetic patients (microalbuminuria, glycated hemoglobin, search for neuropathy, ophthalmology consultation). Interaction with diabetologists, nutritionists and chiropodists was found to be insufficient. Globally, the responses of the diabetics was coherent with the physicians' statements, providing further precision concerning health care delivery and propositions for improvement. These results can be used to better target education programs for associations, specialists and general practitioners. PMID- 11011234 TI - [A public health program for better patient management]. PMID- 11011235 TI - [A public health program for better patient management]. PMID- 11011236 TI - [Epidemiology of diabetes in metropolitan France]. AB - The statistical analysis of French HealthCare's database, containing total reimbursements to affiliates and a listing of delivered drug prescriptions in the form of specific drug codes, allowed us to update data on the epidemiology of diabetes. This study, performed during the first trimester of 1999 on the entire population of metropolitan France, found an overall prevalence of 2.78 % for diabetes in all affiliates of the regimen (Regime general) stricto sensu (3.06 % in the general population), 0.41 % corresponding to diabetic treated exclusively with insulin (0.44 % in the overall population) and to 0.12 % receiving both insulin and oral hypoglycemic agents concomitantly (0.13 % in the general population). The average age of diabetics, irrespective of the treatment they were receiving, was 63.1 years with a sex ratio M/F of 1.04. These findings confirm epidemiological forecasts which had predicted a significant increase in the prevalence of diabetes over the past few years. The study also confirmed data from the medical literature concerning geographic disparities in the epidemiology of this disease except for the results obtained in the north of France, a region generally thought to have a lower prevalence of diabetes, and which was found to have a prevalence equal to even greater than the national average. PMID- 11011238 TI - [Modalities of follow-up on non-insulin treated diabetics treated in metropolitan France in 1998]. AB - The statistical analysis of French HealthCare's database allows a retrospective evaluation of the overall clinical care given to diabetics in the light of the guidelines (RBPC) promulgated by the National Agency of Health Accreditation and Evaluation (ANAES). This database contains no information on the stage of the disease process, the completeness of the clinical evaluation (neurological or foot examination) or the results of significant blood tests. We studied the follow-up care given to diabetics whom we were able to identify by isolating reimbursements for hypoglycemic agents using their corresponding drug codes. Considering the time needed for this new drug-coding system to become fully operational, the results can be extrapolated to the entire population of non insulin treated diabetics. In 1998, almost all follow-up care was provided by general practitioners since only 5.5 % of the patients consulted an endocrinologist during the year studied. Blood sugar was generally monitored with fasting blood sugars rather than HbA1c which was only prescribed in 41.3 % of the patients during the final six months of the period under consideration. Screening and follow-up for degenerative complications were insufficient. Fewer than 40 % of the patients had received reimbursement for an ophtalmological consultation during the preceeding year and fewer than 30 % had an ambulatory resting EKG over the same time period. A certain number of factors were studied in order to determine if they were correlated to better clinical follow-up. The best follow up was encountered in patients who had a consultation with an endocrinologist at least once during the year, those who were exonerated from co-payments (ALD 30) and finally, those aged between 40 and 59 years (at least with respect to appropriate laboratory tests). PMID- 11011237 TI - [The cost of diabetes in metropolitan France]. AB - The statistical analysis of French HealthCare's database, containing total reimbursements to affiliates and a listing of delivered drug prescriptions in the form of specific drug codes, allowed us to update data of the cost of diabetes. This study, realized during the first trimester of 1999, used the combined databases of 110 local healthcare offices (CPAM) among the 128 existing in metropolitan France. French Social Security spent a total of 31.9 billion francs (4.9 billion euros) in care given to diabetics (or 24,137 francs per patient) comprised by 14.2 francs (2.2 billion euros) for in-hospital care and 17.7 billion francs (2.7 billion euros) for outpatient services. The total expenses for diabetic patients represented 4.7% of French Social Security's overall expenses for all its affiliates. The differential cost of diabetes (the difference, at equivalent ages, between cost generated by diabetic patients and those attributable to the remainder of the affiliated population) was estimated, for the generalized health-care regimen (regime general) in 1998, to be 13.3 billion francs (2.0 billion euros). These figures underscore the need to reinforce prevention and screening for end-organ complications since it is generally admitted that the disease's high cost is essentially related to its complications. PMID- 11011240 TI - [Totally reimbursed care for type 2 diabetes in 1994: characteristics and follow up]. AB - Non-insulin-dependent diabetes mellitus (NIDDM) is an important health issue in France. According to projections from current data, the problem should increase over the upcoming years. The aim of this study was to determine the characteristics of patients with NIDDM who are enrolled on the list of 30 long term diseases (ALD30) and examine their follow-up and disease-related complications. This study was performed with the data base established by the French National Health Insurance for Salaried Workers (CMANTS) in 1994. Diagnosis and medical information were coded by official physicians, who either examined the patients or consulted their primary care physician (69% of the cases), or relied on available medical records. Eight thousand two hundred forty-seven patients (sex ration=0.95) with NIDDM enrolled on the long term disease list were included. Mean age was 65.1+/- 0.13 years (m+/- SEM). More than one blood glucose level per year was available for 89% of the patients and an HA1c assay in only 34%. In addition, for patient follow-up at least once a year, 80%, 47%, and 42% of the patients had a serum creatinine assay, a funus examination, or an electorcariogram respectively. Irrespective of the criteria retained (fasting glucose=1.26 g/l; fasting glucose=1.40 g/l) there was a high percentage of patients whose level was well above the limit (75% and 63% respectively). For HbA1c, only a quarter of the patients had values lower than the limits defined by the ANAES and AFFSAPS guidelines. The percentage of diabetic patients without complications was 41% (hypertension excluded). The most frequently observed complications isolated peripheral vascular conditions (31%). These results were obtained from a database created in 1994, before guidelines on type 2 diabetes were officially published. They can serve as a reference to follow medical practices. PMID- 11011239 TI - [Analysis of health insurance benefits: material and methods]. AB - The statistical analysis of national health insurance's database, containing a record of the care reimbursed to affiliated members and delivered drugs in the form of special pharmacy codes, enable us to evaluate the follow-up care given to diabetics, to update epidemiological information and to determine the actual cost of diabetes. The method used consisted in the identification of diabetics by isolating patients who received hypoglycemic agents during the 3-month inclusion period followed by a retrospective determination of total reimbursements made through the 128 local health offices (CPAM) located in metropolitan France in order to study the reimbursed care (medical consultations, procedures and drug prescriptions) given to diabetics over the preceding 12 month period. Considering that the pharmacy coding system was still incomplete at the time the study was undertaken, we had to verify that the population selected by the coding system was only partially operational, requiring a calculation involving weighted coefficients. Finally, we needed to take into account the fact that the resulting data contained virtually no information on facilities receiving a once a year total budget allocation (all public hospital). PMID- 11011241 TI - [Pharmacoepidemiology of diabetes: assessment of good use of oral antidiabetic drugs]. AB - Diabetes' treatment is specific; therefore diabetic patients can be identified by the prescription of antidiabetic drugs. This pathology is characterized by chronicity and occurrence of complications which need additional medications. All the drugs have official use guidelines due to drug interactions and/or physiological status of the patients. To evaluate how these guidelines are followed in general practice, we used a data base from the French Health care System. Two thousand eight hundred and ninety eight (2 898) prescriptions on which, at least, one oral antidiabetic drug was noticed, were analysed. The number of drugs on each prescription was between 1 and 18 (mean (5,6 +/- 2,9)). Considering the drugs which need precautions of use in association with sulfonylureas, coprescription occurred with B-bloquants and non steroid anti inflammatory drugs in 18 and 17 % of the cases respectively. The drugs which need precautions of use in association with biguanides were diuretics, oestrogens and/or progestatives and steroids, coprescribed in 28, 1, 1 % of the cases respectively. Percentages of coprescriptions which need precautions of use were quite high. The relevance of detected coprescriptions. and the compatibility between recommendations for "good prescription" and "real prescription practice" should be taken into account considering diabetic patients' characteristics. PMID- 11011242 TI - [Description of type 2 diabetes mellitus in residents of Ile-de-France aged at most 70 years]. AB - Our study of patients with type 2 diabetes mellitus residing in the Paris area (Ile-de-France) aged 70 years or younger included 1 591 patients examined by national health insurance expert physicians who also filled out a self administrated questionnaire. Their primary care physicians also answered a questionnaire (79% participation, indicating their personal implication). We found that diabetes was most often discovered during regular check-ups. Single drug regimens were the most widely used. Sulfamides were prescribed for 77% of the patients. Among the 912 patients who were overweight (BMI > 28 Kg/m2), 34.3% were taking sulfamides alone despite the fact that metformin is recommended as the first intention drug for these patients. Blood pressure control was not satisfactory in 30% of the patients who were treated or not for high blood pressure. This proportion rose to about 50% among treated patients alone. The patients appeared to be knowledgeable about the risk of complications, particularly ocular complications. They were aware of a certain number of messages but the results would suggest difficulty in application. PMID- 11011243 TI - [Identification of factors hindering better management of patients with type 2 diabetes]. AB - The aim of this study was to help improving management of patients with type 2 diabetes by evaluating their knowledge of the disease and its treatment and by identifying factors which might influence management. A questionnaire was sent to diabetic patients taking oral antidiabetic drugs affiliated to one of the three main national health insurance schemes in the French Departments Seine-Maritime and Eure. Four hundred responses were analyzed. Patient knowledge of diabetes and its complications, follow-up, surveillance and dietary control were rather poor. Use of a personal diary (used by 24%) was associated with better knowledge of the disease and better participation in follow-up and treatment. Actions aimed at better education of diabetic patients promoting use of a personal diary would be helpful to improve disease management, necessary to reduce the risk of complications. PMID- 11011244 TI - [Malignant duodenal stenoses: can endoscopic or radiological insertion of metal stent replace surgical derivation?]. PMID- 11011245 TI - [Iron deficiency in patients over 60 years. Descriptive study in the consultant population of health screening centers]. AB - AIM: The aim of the study was to evaluate the frequency of iron deficiency with serum ferritin in elderly population, and to appreciate the opportunity of early screening according to digestive diseases. SUBJECTS AND METHODS: Data were collected from 3524 men and 3120 women aged 60 to 75 years during a health screening examination. Evaluation of diagnosis and treatment were obtained through questionnaire completed by treating physician. RESULTS: The frequency of hypoferritinemia was about 2.3% in our population (hypoferritinemia was defined by serum ferritin<20 microg/L or between 20-40 microg/L if C reactive protein was > 12 mg/L). Anemia was found in 3.3% of patients. Logistic regression model adjusting for multiple variables was used to examine factors associated with hypoferritinemia. The probability was greater among non-anemic patients with chronic digestive bleeding (odds-ratio: 2.3), or with positive occult blood testing (odds-ratio: 2.3). Information about the medical follow-up was obtained in 81% of patients with hypoferritinemia. A digestive exploration was made in 38 cases. Digestive disease was found among 24.3% patients with hypoferritinemia, and three colorectal cancers were observed. CONCLUSION: The screening of hypoferritinemia in elderly population examined in health screening centres could not be recommended as its frequency was low in this population, despite a strong correlation between hypoferritinemia and digestive diseases. PMID- 11011246 TI - [Palliative endoscopic treatment of malignant duodenal stenosis by metal prosthesis]. AB - PURPOSE: To evaluate the efficacy of through-the-scope metal stents for palliation of malignant duodenal stenosis. MATERIAL AND METHODS: Fourty two patients with malignant primary or secondary duodenal stenoses who were treated with a through-the-scope metal stent were analysed. When obstructive jaundice occurred either before, during, or after the initial episode of gastrointestinal luminal obstruction, a biliary stent was inserted. RESULTS: Duodenal metal stents were deployed in 40 patients. Endoprosthesis insertion led to restoration of oral intake in 39 patients. The procedure was not associated with morbidity or mortality. During a mean follow-up of 9.7 weeks, adequate oral intake was maintained in 38/39 cases. Tumour in-growth led to stent occlusion in 4 cases and re-cannulation was obtained by placement of another stent within the original stent. Obstructive jaundice occurred during the course of the illness in 32 patients and was successfully treated with a biliary metal stent in all cases. CONCLUSIONS: Endoscopically placed metal stents offer an effective, well tolerated alternative to surgical palliation in case of incurable malignant obstruction to gastric outflow. PMID- 11011249 TI - [Pre-test]. PMID- 11011247 TI - [Peroxisome proliferator-activated receptor (PPAR) gamma: a new target for the treatment of inflammatory bowel disease]. AB - The peroxisome proliferator-activated receptor (PPAR) gamma is highly expressed in the colon mucosa. In vitro, it regulates inflammation. AIM: To evaluate the anti-inflammatory functions of PPARgamma agonist during a trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. METHODS: Colitis was induced in Balb/c mice after intra-rectal administration of TNBS. The intensity of inflammation was assessed 2 and 5 days after colitis induction by macroscopic and histologic scores and by the quantification of colon myeloperoxidase (MPO), IL-1B and TNFalpha mRNA concentrations. The therapeutic role of PPARgamma agonist given by oral gavage was assessed in preventive and treatment modes. RESULTS: TNBS induced severe macroscopic and histologic lesions, with high mucosal MPO, IL-1B and TNFalpha mRNA concentrations. PPARgamma agonist given preventively or in treatment mode allowed a significant decrease of macroscopic and histologic scores through a normalization of MPO, IL-1B and TNFalpha mRNA concentrations. CONCLUSION: PPARgamma agonist decreases the intensity of TNBS induced colitis through normalization of IL-1B and TNFalpha expression. PPARgamma agonists may be proposed as new therapeutic agents in inflammatory bowel diseases. PMID- 11011248 TI - [Natural history of chronic alcoholic pancreatitis]. PMID- 11011250 TI - [Angina-like chest pain: physiopathology, diagnostic measure and treatment]. PMID- 11011251 TI - [Chest pain from hypertrophic esophageal muscles]. PMID- 11011253 TI - [Answers to the pre-test]. PMID- 11011252 TI - [Interview: questions to Pr Jean-Paul Galmiche]. PMID- 11011254 TI - [Biliary prosthesis and malignant obstruction of extraheptic bile ducts]. PMID- 11011255 TI - [Pegylated interferon: new progress in treatment for chronic hepatitis C]. PMID- 11011256 TI - [Malignant obstructive jaundice: the role of percutaneous metallic stents]. AB - OBJECTIVES: Efficiency evaluation of percutaneous metallic stents in palliative treatment of malignant biliary obstruction. METHODS: One hundred sixteen percutaneous metallic stents were implanted in 80 patients with malignant biliary obstruction. Thirty-five patients had hilar obstruction, 32 patients obstruction of the common bile duct and 12 patients obstruction of a bilioenteric anastomosis. RESULTS: Adequate biliary drainage was achieved in 79 patients. Early complications occurred in 23.75% of patients; 12.5% of patients died within 30 days. The procedure-related mortality rate was 5%; 18.75% of patients showed recurrent jaundice after an average of 175 days. CONCLUSION: Percutaneous metallic stents are an efficient means of treating malignant biliary strictures, particularly of upper biliary obstructions. However, this treatment has risks and limits that require careful patient recruitment. PMID- 11011257 TI - [Evaluation of nursing care during treatment of viral hepatitis C by interferon]. AB - OBJECTIVE: Interferon alpha treatment requires parenteral administration and some patients receive injections from nurses. The aim of this study was to evaluate the role of nursing care during ambulatory treatment of hepatitis C by interferon alpha. METHODS: A questionnaire was sent by mail to all nurses who practice ambulatory care in the Val de Marne region (n =545). RESULTS: One hundred fourteen questionnaires were returned (return rate: 20%). Among these nurses, only 42 (37%) were caring for at least 1 patient for the treatment of hepatitis C by interferon. In the 16 months before the survey, these 42 nurses had cared for 135 patients. The nurses cared for the patients throughout the entire treatment in 81% and only to teach them in 19% of cases. The time spent for each injection was 12 min 48 sec, 2 min 42 sec for the injection itself, 3 min 54 sec to answer to patient's questions, 3 min 30 sec for psychological support, and 2 min 48 sec for unspecific time. In relation to hepatitis C and interferon treatment, 74% and 64% of the nurses felt that patients asked questions they did not ask their physician. Seventy seven of the nurses felt they were did not have enough training to answer the questions. Psychological support for the patient was noted as important or very important by 38 % of nurses. CONCLUSIONS: The role of nurses during interferon treatment is not limited to injecting the drug. Advice and psychological support takes up most of the time spent with patients. Specific training and better collaboration with the physician who prescribes the treatment are needed. PMID- 11011258 TI - [Living donor in liver transplantation. Present and future alternatives]. PMID- 11011259 TI - [Yellow nail syndrome associated with intestinal lymphangiectasia]. AB - The yellow nail syndrome, a combination of yellow discoloured nails, lymphedema and pleural effusions, is a rare clinical condition. We report a case of the yellow nail syndrome associated with intestinal lymphangiectasia revealed by chylous ascites and protein-losing gastroenteropathy. This association reported in only three cases in the literature leads us to discuss the relations between yellow nail syndrome, primitive intestinal lymphangiectasia and primary lymphatic disorders. PMID- 11011260 TI - [Peritonitis with Mycobacterium avium complex in a patient with alcoholic cirrhosis]. AB - Before highly active antiretroviral therapy were available, disseminated Mycobacterium avium complex infection was common in adults with HIV. Diagnosis was often made by blood culture in these immunocompromised patients. Although Mycobacterium avium complex disease can involve any organ of the body, infection of serosal surfaces is very rare. Mycobacterium avium complex peritonitis is rare and usually occurs in immunocompetent patients with chronic ambulatory peritoneal dialysis. We report a case of Mycobacterium avium complex peritonitis in a patient with alcoholic cirrhosis with no evidence of HIV infection. Diagnosis was made by culture of a lymphocytic ascites which showed Mycobacterium avium complex at 4 weeks. Interestingly, blood and hepatic cultures remained negative. At three months, marked improvement occurred with antimycobacterial treatment, so that orthotopic liver transplantation could be performed. PMID- 11011261 TI - [Primary T-cell lymphoma of the common bile duct]. AB - Involvement of the gastrointestinal tract is frequently reported among the extranodal sites of non-Hodgkin's lymphoma, but primary lymphoma of the common bile duct is extremely rare. We report the case of a 29-year-old man who presented with obstructive jaundice, leading to the diagnosis of high-grade primary non Hodgkin's T-cell lymphoma, originating from the extrahepatic biliary tract, and confirmed by endosonography and magnetic resonance cholangiography. This patient was treated by sequential chemotherapy without resection and remained in complete remission after one year. PMID- 11011262 TI - [Bronchiolitis obliterans with organized pneumonia and hemorrhagic rectocolitis]. PMID- 11011263 TI - [Gastric heterotopia in the rectum: a new case]. PMID- 11011264 TI - [Delayed urticaria with oxaliplatin]. PMID- 11011265 TI - [Lymphocytic colitis due to Modopar]. PMID- 11011266 TI - [Interferon and cryoglobulinemia: how to inject without precipitation]. PMID- 11011267 TI - [Digestive endoscopy aanesthesia: data from the SFAR]. PMID- 11011268 TI - [Reply from Remy Dumas to Andre Lienhart]. PMID- 11011269 TI - [Office-based colonoscopy with possible sedation as requested performed by the endoscopist: a method also possible in France]. PMID- 11011270 TI - [Reply from Remy Dumas to Bernard Maroy and Philippe Moullot]. PMID- 11011271 TI - [Reply from Denis Grasset, Christian Seigneuric and Jean-Jacques Morfoisse to Bernard Maroy and Philippe Moullot]. PMID- 11011272 TI - [Prevention of hemolytic disease of the fetus and the newborn: it is necessary to act]. PMID- 11011273 TI - Echographic signs of trisomy 21 in the second trimester of pregnancy: actual value after analysis of the literature. AB - OBJECTIVE: To evaluate the interest and to compare the major echographic signs of Down syndrome in the second trimester of pregnancy. METHODS: A bibliographic research has been performed for most of the echographic signs known and studied until now. For each study and in average for each sign, we have computed its sensitivity, its specificity, its positive and negative predictive values using the results of the different authors. Then, we have compared the benefits/risk ratio for each of these signs: the number of Down syndrome cases detected versus healthy fetus lost due to amniocentesis complications. RESULTS: The different signs can be ranked according to their benefits/risk ratio from top to bottom as follows: nuchal skinfold thickness, wide space between first and second toe, pyelectasis, large iliac angle, short humerus, short femur, hypoplasia of the middle phalanx of the fifth digit. CONCLUSION: These results suggest that second trimester echographic signs of Down syndrome must be evaluated as a function of the Down syndrome risk in the population under study. The presence of these signs does not always justify an amniocentesis; it should lead to a re-evaluation of the individual risk of a Down syndrome (a chart is given to guide this re evaluation). PMID- 11011275 TI - [Repetition of corticoid treatment for fetal lung maturation: clinical and experimental scientific data]. AB - Before 34 weeks, one course of antenatal corticosteroids significantly decreases neonatal mortality and morbidity. Courses are repeated every week, because of their alleged loss of efficiency after 7 days, although the relative benefits and risks of repeated courses are poorly known. Data from the literature suggest that improvement in lung function after repeated courses depends on the reduction of the treatment/birth interval and not on the rise in the cumulative dose. These benefits must be balanced against the risk of decreased birth weight induced by repeated courses. Because of their effect on fetal growth, weekly courses of corticosteroids should no longer be given systematically. In clinical practice, pregnancies at high risk of preterm delivery must be reassessed every week before prescribing a new course. Both better understanding of treatment duration of corticosteroids and randomized controlled trials comparing one and multiple courses are needed to improve antenatal management of neonatal respiratory stress syndrome. PMID- 11011274 TI - [Parkinson's disease and pregnancy: case report and literature review]. AB - Parkinson's disease begins before the age of 40 in only 5% of patients. We report a case of pregnancy in a 35-year old woman with Parkinson's disease. The patient was treated by levodopa associated with benzerazid. No effect was observed on the pregnancy and the fetus. Only the symptoms were moderately worsened. A literature review is performed. PMID- 11011276 TI - [Infraclinical breast lesions: apropos of a retrospective study of 261 cases]. AB - OBJECTIVE: To determinate the rate of malignant lesions discovered after surgery for nonpalpable breast lesions diagnosed by radiology. TOOLS AND METHODS: We performed a retrospective study of 261 patients who underwent surgery for nonpalpable breast lesions at the Saint-Etienne University Hospital between 1991 and 1998. We included women who did not have any palpable breast lesion. We excluded bilateral lesions and nonpalpable lesions associated with a palpable lesion in the same breast. We grouped outcome by alarming signs, those of complementary exams such as mammography, ultrasound, fine-needle aspiration biopsy and setreotaxis core needle breast biopsy in their sensitivity, specificity and predictive values, and classed results by pathology and type of surgery performed. All nonpalpable lesions were localized by stereotaxis maneuvers. RESULTS: In our series, 61.7% of the patients were seen for individual detection, 3% had participated in mass screening programs and 35.3% were referred for suspected conditions. The predictive value of malignant lesions on mammography, analyzed in all cases and interpreted in 275, was 42.2%. The sensitivity and specificity of ultrasound exams performed in 50.2% of the cases were 85% and 91% respectively. Fine-needle aspiration biopsy and core needle biopsy were performed in 45 (18%) and 23 (9%) cases respectively. Twelve patients underwent total mastectomy with axillary dissections, 58 had tumorectomy with axillary dissections, and 191 had simple tumorectomy. Benign lesions were found in 128 (49.1%) cases, atypical hyperplasia in 10.7%, carcinoma in situ in 57 (cases 21.8%). The rate of malignant lesions was 38.6%. I was 50.9% when borderline lesions (atypical hyperplasia and in situ lobular carcinoma) were included. CONCLUSION: In our series, 38.6% of all nonpalpable operated breast lesions were cancers. Ultrasound, fine-needle aspiration and needle core biopsy showed high sensitivity and specificity allowing good biopsy outcome. PMID- 11011277 TI - [Complications of vaginal hysterectomy on non-prolapsed uterus]. AB - OBJECTIVE: To assess our rate of complications after vaginal hysterectomy and those described in the literature, particularly in high risk groups such as obese, nulliparous women with previous abdomino-pelvic surgery or after GnRH administration. METHOD: We retrospectively looked for complications in 490 patients who underwent vagina l hysterectomy between 1990 and 1998: infections, hemorrhage, intraoperative complications, thrombophlebitis or pulmonary embolism, postoperative complications up to one month. The chi-squared test, t test for equality of means and Fisher's exact test were used for statistical analysis. RESULTS: The complication rate observed in our series (20.7%) was similar to those reported in the literature. We found no difference in the obese group (n=109) the nulliparous women (n=34), after GnRH administration (n=24) or i women with previous abdomino-pelvic surgery (n=128). Heavy bleeding was not increased by morcellation. CONCLUSION: Vaginal hysterectomy is easy and safe. The vaginal route is the best way for obese subjects and can be carried out in most cases, especially in nulliparous women or after GnRH analogs, even when morcellation is necessary, without increasing morbidity. PMID- 11011278 TI - [Regret after tubal sterilization]. AB - OBJECTIVE: To examine thoroughly the profile and motivations of a group of women who regretted sterilization so much that they were prepared to undergo reversal microsurgery or in vitro fertilization treatment. MATERIAL: and methods. The study followed one hundred women from their first consultation for a period of 3 years in order to analyze the outcome of their initial decision. RESULTS: Analysis of the circumstances at the time of the sterilization indicate 3 main factors which later caused the women to regret the decision Three years after the initial consultation, 69% of the patients have withdrawn from further investigation and /or treatment for different reasons analyzed in this study. 17 of the patients have undergone reversal microsurgery and 14 have attempted one or more in vitro fertilization treatments. Of these 31 patients, 12 (38.7%) have given birth to at least one child. CONCLUSION: These results sufficiently prove that tubal sterilization is worthy of consideration as much by the patient as by the physician as a method of irreversible contraception despite the developments in tubal microsurgery or in medically assisted procreation. PMID- 11011279 TI - [Prenatal screening for trisomy 21 with maternal serum markers: information for decision-making in pregnant women]. AB - INTRODUCTION: French health professionals are now required to provide relevant information on the test screening for Down syndrome and its possible implications when making the test available to pregnant women. A questionnaire was designed to assess prescription modalities, explanation of results, opinion on the information received when test was offered and test results were given, and likely decision after screening. RESULTS: The response rate was 39% (1473 questionnaires analyzed). In 90.5% of the answers, women declared that an interview had preceded test prescription. 61.2% of women were satisfied with both the clarity and quantity of explanations provided and 57.6% considered that the information had helped them decide to undergo or refuse screening. 54.1% were satisfied with the explanations of test results. Amniocentesis was proposed to 125 women, 79.2% of which decided to undergo the procedure. Their opinion concerning explanations of test results was similar to that of women who refused amniocentesis or had not yet made up their minds. CONCLUSION: Such results demonstrate the necessity to improve and adapt the information delivered by health professionals, keeping in mind that in this type of situation, decision making requires the rapid integration of complex information. PMID- 11011280 TI - [Maternal mortality in the maternity ward of Befelatanana. University Hospital of Antananarivo]. AB - OBJECTIVE: A retrospective study was conducted to assess mortality rates at the Befelatanana Women's Hospital from January 1st 1988 through December 31st 1997, define trends, study causes and identify problems encountered during treatment and those related to the referral system, in order to improve prevention and patient management MATERIAL: and method. Birth and death registries, operation reports and patient charts were analyzed. All maternal deaths wether of direct or indirect obstetrical origin were considered. RESULTS: There were 997 cases of maternal death among 91, 032 births, i.e. 1.09% or 1,245 maternal deaths per 100,000 live births. Among direct obstetrical causes of death (93%), 65.8% involved infectious complications and 22.9% renal and vascular complications.Other causes were anesthesia accidents and thrombolic events. Among the direct obstetrical causes, complicated delivery was responsible for 57.4% of the deaths and 42.6% of the abortions. Indirect obstetrical causes were due to malaria (68.6%), heart disease (21.4%) and asthma (10%). DISCUSSION: This high rate of maternal death illustrates the serious situation of obstetrics in our region. Our aim is to reduce this rate by promoting the Birth Place Without Risk program and improving management conditions. Implementation of this ambitious program requires implication of political decisions makers convinced of the importance of improving women's conditions and the health care system in Madagascar. PMID- 11011282 TI - [Waldmann disease and pregnancy]. AB - Waldman disease, also called intestinal lymphangiectasis, is a protein-loosing enteropathy caused by anomalous intestinal lymphatic canals. The condition is transmitted by autosomal dominant inheritance. Diet therapy including medium chain triglycerides improves the disease course. The condition occurs in early childhood. We report a case observed in a pregnant woman with intestinal lymphangiectasis. She delivered a full-term infant and experienced no major complications. PMID- 11011281 TI - [Determination of acceptability of 2 cutaneous estradiol gels, in a dose of 1.5 mg daily]. AB - OBJECTIVE: The aim was to compare acceptability of a percutaneous 0. 1% estradiol gel (Gel A, Estreva(R) Gel, Laboratoire Theramex, Monaco) to that of an 0.06% estradiol gel (Gel B, Oestrodose(R), Laboratoires Besins-Iscovesco) in its new formulation and packaging. MATERIAL AND METHODS: This randomized, crossed, simple blind study was carried out in 48 volunteer healthy postmenopausal women. The volunteers applied on one forearm 1.5 mg/day of cutaneous estradiol in the form of either gel, according to randomized allocation, for four days without free period between the two therapeutic periods. The application and drying times of the two gels were measured during the first application; gel subjective women assessment was collected at the beginning and at the end of the study. RESULTS: Mean application and drying times with Gel A are significantly reduced, compared to Gel B (p=0.0259 and p=0.0001, respectively) with drying time 61% shorter; these data are confirmed by subjective women evaluation. The two gels are not significantly different regarding several criteria as consistency, ease of application and sensation of lasting stickiness. However, a significant difference is found in favour of Gel A on the following items: practicality of application (p=0.007), ease of penetration (p<0.001), quantity of gel to apply (p<0.001) after the first application. After four days of administration, a same significant difference is observed concerning practicality of the gel (p=0.0078), duration of use (p<0. 001), packaging, women opinion on the gel (p=0.022) and the product, gel and packaging (p<0.001). At the end of the study, gel A utilization is considered by women more practical (p=0.001) with an easier application (p<0.001) and less restricting while applying (p=0.001), compared to Gel B; 72.9% of women prefer the Gel A and 12. 5% of women prefer the Gel B. CONCLUSION: A better acceptability of the 0.1% estradiol gel and of its packaging compared to that of the 0.06% estradiol gel in this new formulation and packaging is observed in this study. PMID- 11011284 TI - [In Process Citation] PMID- 11011283 TI - [Patient information: responsibility of all physicians to provide proof (hospital as well as private)]. PMID- 11011285 TI - [Breech presentation: delivery]. PMID- 11011298 TI - [MONICA Project. At the heart of the epidemiological question]. PMID- 11011299 TI - Mammographic screening in women with a family history of breast cancer: some results from the Swedish two-county trial. AB - BACKGROUND: The objective of this study is to compare the effectiveness of mammographic screening in women with a family history of breast cancer to those without. In the invited arm of a randomised trial of breast cancer screening, data on family history of breast cancer were available on 29.179 women aged 40-74 attending for screening. Among those women, 358 were diagnosed with breast cancer during the trial. METHODS: Those with and without a family history were compared with respect to mammographic parenchymal pattern, interval cancer rates, mean sojourn time and sensitivity of screening. In the 358 cancers, the effect of family history was estimated on survival, incidence of advanced cancers and their relationship to screen detection. RESULTS: A significantly higher proportion of high risk mammographic patterns was observed in association with family history among women aged 40-49. Interval cancer rates were higher in women with a family history, and in older women at least, mean sojourn time was shortened in women with a family history (1.89 years compared to 2.70). Survival was better (although not significantly so) in cancers in women with a family history (relative hazard=0.52) independently of detection mode and was significantly poorer in interval cancers then screen detected cancers (relative hazard=2.72) independently of family history. Similarly, interval cancers tended to be larger, and worse malignancy grade in those with and without a family history of breast cancer. CONCLUSIONS: These results suggest that the policy often adopted of annual screening for woman aged 40-49, with a family history of breast cancer, is a reasonable one, and that it may also be necessary to shorten the inter screening interval to one year in women aged over 50 but with a positive family history. PMID- 11011300 TI - [Breast cancer screening and medical consumption]. AB - BACKGROUND: The use of general health services by women targeted by breast cancer screening campaigns, and its variations, are poorly known. A survey has been undertaken during one of these campaigns in a britton department. This study aimed at pointing out possible differences in use of general health services: search for an overuse by women whose screening resulted positive/negative, in relation to women whose screening resulted full negative; search for an overuse by women having participated, in relation to women within the age range of the screening campaign and not having wished to participate. METHODS: Three groups of women, aged 50 to 69 years, free of breast cancer, were compared. A random sample was drawn in each group, weighing for age in group D0. Inclusion of cases was undertaken between January 1995 (beginning of the screening campaign) and June 1996. Use of health services was recorded during the six months following the first screening test, using the health insurance services data. Expenses related to inpatient care were not recorded. RESULTS: The structure of age of the three groups was comparable. The proportion of women having requested no reimbursement was identical between the groups D+ and D-. For the D+ group, the average cost was 4,246 Francs per woman. It dropped to 3,871 Francs after deduction of expenses related to additional medical examinations following the finding of a positive screening test. It was 5% higher than the average cost in the D- group (3,698 Francs). The cost varied a great deal within each group and the differences observed between the groups were not statistically significant The proportion of women of the D0 group not having requested any reimbursement was significantly lower than those of the groups D- and D+, overall, and for the majority of the categories of expenditure. CONCLUSION: The result of the screening does not induce significant variation of use of general health services, apart from the expenses induced by the invalidation of the positive result. The group of women not having participated in screening includes a stronger proportion of both low and higher health services users. PMID- 11011301 TI - [Personal exposure to fine particles (PM 2.5) in the Grenoble population: European EXPOLIS study]. AB - BACKGROUND: The aim of this article is to describe PM(2.5) personal exposures within the Grenoble population and to identify the implications of such measurements on epidemiological or risk assessment studies of air quality. METHODS: Non smoking adult volunteers, selected in summer 1996 (n=40), in winter 1997 (n=40) and in summer 1998 (n=20), carried a case containing 2 PM(2.5) personal monitors. One of the monitors was running continuously for 48h. (cumulative exposures), the other one was running only while indoors (indoor exposure). PM(2.5) masses were determined by reflectometry (black smoke method) and by deionised weighting (Mettler MT5 micro-balance; gravimetric method). RESULTS: Cumulative PM(2.5) personal exposures ranged on average from 21.9 in summer to 36.7 microgram/m(3) in winter (arithmetic mean), using the gravimetric results; the dispersion of these personal exposures was greatest in winter (s.d.=23.1 microgram/m(3)) than in summer (s.d.=10.4 microgram/m(3)). There was a good correlation (R=0.7) between the reflectometry and weighting results in winter, but not in summer. Outdoor personal exposures, determined by difference between the cumulated and indoor masses, were slightly higher than the cumulative personal exposures: the geometric means was 49.6 microgram/m(3) (geometric standard deviation=2.7 microgram/m(3)) in summer and 55.1 microgram/m(3) (3.7 microgram/m(3)) in winter (gravimetric results). Due to these greater outdoor concentrations, the fraction of outdoor exposure was high (25%) relative to the small amount of time spent outdoors (less than 10%). CONCLUSION: These descriptive data, consistent with the literature, show the importance of "expology" studies aiming at characterizing PM(2.5) personal measurements across the year. This would lead, in the future, to optimizing the use of "ecological" estimates of exposures from ambient air concentrations provided by the ambient air quality networks, for the characterization of exposure in epidemiological or risk assessment studies. PMID- 11011302 TI - [Psychosocial distress and its moderating factors in patients living in precarious socioeconomic conditions consulting in a preventive health and social work center]. AB - BACKGROUND: The effects of a precarious socioeconomic condition on mental health have already been widely described. Nevertheless, sociodemographic, biographical, and mainly psychological determinants of psychosocial distress in these populations are still incompletely known. METHODS: 2315 consecutive subjects, aged 16 to 59, consulting for a free work-up in a preventive health center supported by the National French Health Insurance system, were invited to fill out a series of questionnaires, mainly the GHQ-28 (psychosocial distress), the LOT (dispositional optimism) and the WCCL (coping mechanisms). Socioeconomic and administrative criteria were adopted to define populations living in precarious conditions. RESULTS: A total of 78.9% complete records could be analyzed: 55% of the studied population constituted the precarious condition group (44.5% males and 55.5% females; mean age 36.2+/-11). This group was characterized by higher psychosocial distress and higher sub-scores of anxiety, social dysfunction and depression, but not somatisation, as well as by increased emotion focused coping and impaired optimism. Globally, GHQ score was positively correlated with emotion focused coping (r=0.36) and negatively with problem focused coping (r=-0.17) and with optimism (r=-0.39). In a multivariate analysis a set of 9 independent variables explained 38% of GHQ-28 total variance: perceived stress, optimism, emotion focused coping, problem focused coping, age, educational degree, precariousness, money problems and parental quarrels during youth. Introducing precariousness in the model constituted of the set of the remaining variables could explain only 2% of additional variance. CONCLUSION: These results confirm the presence of higher levels of psychosocial distress in subjects living in precarious conditions and underline the moderating role of several psychological variables. They could guide support interventions aimed at helping subjects living in precarious conditions, focused on enhancing their coping resources. PMID- 11011303 TI - [Two years follow-up of a heroin users cohort treated with high dosage buprenorphine. Results of the SPESUB study (pharmacoepidemiologic follow-up of general practice Subutex)]. AB - BACKGROUND: Since February 1996, French GPs are allowed to prescribe high dosage buprenorphine for maintenance treatment of major opioid drug addiction. A prospective cohort of major opioid addicts was initiated in order to assess patient outcomes: follow-up, retention rate in treatment, drug use, intravenous injection and social situation evolution. METHODS: Each GP, known to be involved in drug user management, had to include the first 10 opioid drug addict patients to whom he prescribed high dosage buprenorphine, with a maximum inclusion period of 3 months. Patients were followed up for two years and a regular standardized information was collected (usual data on drug users and prescription modalities). RESULTS: Between May and July 1996, 919 patients (664 men and 255 women, mean age: 30 years) were included by 101 GPs. They had a long and serious history of drug addiction, important parallel consumption of cocaine, codeine and other illicit drugs and psychiatric problems (28% of definite problems and 45% of probable) and frequent hepatic conditions (hepatitis B: 23%, hepatitis C: 21%). Two years later, 55% of patients were still followed-up by the same GP and an additional 12% were followed by another GP or in a health care service (hospitalized or receiving methadone in a specialized centre). 13% were not followed, but GPs were able to describe their situation. 8% had been included by GPs who had dropped the study. Finally, 12% of patients were lost to follow-up. Among the 508 patients still followed-up by the same GP after 2 years, the substitution treatment rate was 84%. The dosage bracket had widened (inclusion: mean dosage=7.8 mg +/-4.5, minimum=0.8, maximum=28, median=8; after 2 years: mean=7.6 mg +/-5.4, minimum=0.4, maximum=28, median=8) and the duration of the prescription and dispensing had increased. Declaration of heroin intake in the previous month had fell from 40% to 11% and declaration of drug intake from 53% to 20%. Social situation had improved on average (housing conditions and work). There were 12 seroconversions for hepatitis B, 21 for hepatitis C and 4 for HIV. 14% of patients had declared intravenous injection of high dosage buprenorphine in the previous month. CONCLUSION: After two years of follow-up, 55% of patients were still followed-up by the same GP and an additional 12% was followed by another GP or in a health care service. Among patients still followed up by the same GP, a reduction of drug related harm (seroconversions for hepatitis B, hepatitis C and HIV) was observed. PMID- 11011305 TI - [Modeling incomplete observations]. AB - Incomplete observations, common in epidemiology as in many other fields, lead to problems of bias, precision and power. Using a simple example with 3 binary variables, we discuss situations where the observed odds ratio is biased. We present and compare the main strategies of analysis: complete observations modeling, missing data indicator, weighted analysis, simple imputation, multiple imputation, selection models, shared variable models. PMID- 11011306 TI - [Lot quality assurance sampling: methods and applications in public health]. AB - Lot Quality Assurance Sampling (LQAS), developed to meet industrial quality control needs, has been applied to health surveys. The WHO used this method to assess immunization coverage. The sampling strategy was developed to classify lots as acceptable or unacceptable. Lot sampling is an efficient, simple and time efficient procedure for quality assurance. Under certain conditions, efficiency can be improved with double sampling. We describe the method and its theoretical basis and illustrate applications of LQAS in epidemiological surveillance and quality control of medical records. The advantages and disadvantages of this method are presented. PMID- 11011304 TI - [Occupational exposure to organic solvents and pregnancy. Review of current epidemiologic knowledge]. AB - The findings and methodological issues of recent epidemiological studies (1989 1999) on adverse developmental effects of maternal occupational exposure to organic solvents are reviewed. Several studies suggest the possibility of a moderate increase in the risk of spontaneous abortion and congenital malformations, especially facial clefts, associated with maternal exposure to solvents. A common methodological weakness of these studies is the inaccuracy of data on exposures. Positive findings encourage further epidemiological studies of high-quality design and use of protective measures to minimize exposure to these agents from the preconceptional period to the end of pregnancy. PMID- 11011307 TI - [Cardiovascular risk factors among school children in Sousse, Tunisia]. PMID- 11011308 TI - Genetics of coronary heart disease: current understanding and future prospects. PMID- 11011309 TI - Recommendations for national and local regulatory authorities concerning research in genetic markers of disease. PMID- 11011310 TI - Ethical and legal issues in genetic testing to predict risk of heart disease. PMID- 11011311 TI - Genetics of coronary heart disease: current knowledge and research principles. PMID- 11011312 TI - Left atrial enlargement and atrial fibrillation. PMID- 11011314 TI - Algorithms or nomograms for the busy physician? PMID- 11011316 TI - Role of salt in hypertensive left venticular diastolic function. PMID- 11011318 TI - Discrepancy between noninvasive and invasive measurements of MVA after BMC. PMID- 11011319 TI - Aspirin for the treatment of pulmonary embolism: vasoconstriction versus physical obstruction. PMID- 11011321 TI - Genetic approaches for the investigation of genes associated with coronary heart disease. PMID- 11011320 TI - Parasympathetic dysfunction as a risk factor in myocardial infarction: what is the treatment? PMID- 11011322 TI - Genetic analysis for common complex disease. PMID- 11011323 TI - Genetic susceptibility to cardiovascular diseases. PMID- 11011324 TI - Genetics, genomics, proteomics: what is a cardiologist to do? PMID- 11011325 TI - The mode selection trial (MOST) in sinus node dysfunction: design, rationale, and baseline characteristics of the first 1000 patients. AB - BACKGROUND: More than 200,000 permanent pacemakers will be implanted in the United States in 2000 at a cost of more than $2 billion. Sick sinus syndrome (SSS) will likely account for approximately half of all cases necessitating implantation. Pacemaker technology permits the selection of ventricular (single chamber) or dual-chamber devices. However, clinical and outcomes data are inadequate to support a clear recommendation that one or the other type of device be used. METHODS: The Mode Selection Trial (MOST) is a single-blind study supported by the National Heart, Lung, and Blood Institute designed to enroll 2000 patients with SSS. All patients will receive a DDDR pacemaker programmed to VVIR or DDDR before implantation. The average time of follow-up will be 3 years. MOST has a >90% power to detect a 25% reduction in the primary end point-nonfatal stroke or total (all cause) mortality-in the DDDR-treated group. Secondary end points will include health-related quality of life and cost effectiveness, atrial fibrillation, and development of pacemaker syndrome. Prespecified subgroups for analysis will include women and the elderly. Enrollment was completed in October 1999, with a total of 2010 patients. RESULTS: The median age of the first 1000 enrolled patients is 74 years, with 25% of patients 80 years or older. Women comprise 49%, and 17% are nonwhite, predominantly black (13%). Before pacemaker implantation, 22% of patients reported a history of congestive heart failure, 11% coronary angioplasty, and 25% coronary bypass surgery. Supraventricular tachycardia including atrial fibrillation was present in 53% of patients. A prior stroke was reported by 12%. Antiarrhythmic therapy was in use in 18% of patients. CONCLUSIONS: MOST will fill the clinical need for carefully designed prospective studies to define the benefits of dual-chamber versus single-chamber ventricular pacing in patients with SSS. The MOST population is typical of the overall pacemaker population in the United States. Thus the final results of MOST should be clinically generalizable. PMID- 11011326 TI - Extending the useful life of clinical trials. PMID- 11011327 TI - Detection of threatening atherosclerotic plaque: technologic advancement and opportunities. PMID- 11011328 TI - Projected long-term costs of coronary stenting in multivessel coronary disease based on the experience of the Bypass Angioplasty Revascularization Investigation (BARI). AB - BACKGROUND: Stents are now used in the majority of percutaneous coronary revascularization procedures. It is not clear whether the higher initial cost of stenting is later repaid by reducing costly complications and repeat revascularization procedures, especially for patients with multivessel disease. METHODS: To project the long-term costs of using coronary stents, angioplasty, or bypass surgery to treat patients with multivessel coronary artery disease, we developed a decision model based on the outcomes documented in the Bypass Angioplasty Revascularization Investigation (BARI) randomized trial of coronary artery bypass grafting (CABG) and percutaneous transluminal coronary angioplasty (PTCA). We studied 2 clinical strategies: provisional stenting of suboptimal PTCA results and primary stenting of all angiographically eligible lesions. The cost of CABG was also updated to reflect contemporary practice. RESULTS: Provisional stenting had lower projected costs over a 4-year period than either traditional PTCA (-$1742, or -3.4%) or contemporary CABG (-$832, or -1.7%), mostly because of reductions in emergency CABG after PTCA. In contrast, primary stenting had higher projected costs over a 4-year period than either PTCA (+$333, or +0. 7%) or contemporary CABG (+$1243, or +2.5%), mainly because of the higher initial procedure costs. These results were not substantially altered when we systematically varied the key parameters of the models in 1-way and 2-way sensitivity analyses. CONCLUSIONS: A primary stenting strategy in patients with multivessel disease has higher projected long-term costs than CABG. In contrast, a provisional stenting strategy in multivessel disease has lower projected costs than either PTCA or CABG. PMID- 11011329 TI - Tissue characterization of atherosclerotic plaques by intravascular ultrasound radiofrequency signal analysis: an in vitro study of human coronary arteries. AB - BACKGROUND: Conventional gray-scale images of intravascular ultrasound (IVUS) cannot accurately differentiate histologic subtypes of sonolucent coronary plaques with or without a lipid core. METHODS: We analyzed radiofrequency signals obtained in vitro from 24 regions of interest (ROI) of noncalcified (sonolucent) plaques in 10 atherosclerotic coronary artery specimens pressure-fixed by formalin. Radiofrequency signals were sampled with a 30-MHz IVUS catheter and digitized at 500 MHz in 8-bit resolution. The ROIs were histologically categorized into 12 plaques with a lipid core and 12 plaques without it. Integrated backscatter and statistical parameters of the radiofrequency envelope (mean/SD ratio [MSR], skewness, and kurtosis) within the ROI were calculated offline, and their ability to detect a lipid core was compared with visual analysis of the IVUS video images. In the group with lipid cores, percent area of a lipid core in each ROI was measured in a digitized histologic image by a computerized planimeter. RESULTS: Sensitivity and specificity of MSR, skewness, and kurtosis for lipid core detection were substantially greater than visual video image analysis (83.3% and 91.7%, 100% and 91.7%, 100% and 91.7% vs 53.3% and 71.7%). Furthermore, the parameters of integrated backscatter, MSR, skewness, and kurtosis were significantly correlated to percent of core area (r = -0.64, 0.73, 0.78, and 0.63, respectively; P<.05). CONCLUSIONS: Compared with IVUS video images, the parameters of radiofrequency signal analysis may be used to aid in more accurate detection and quantitative evaluation of a lipid core, which is one of the major factors of a vulnerable coronary plaque. PMID- 11011330 TI - Cardiac structure and function in fetuses of mothers infected with HIV: the prospective PCHIV multicenter study. AB - BACKGROUND: This study was designed to determine if vertically transmitted HIV infection and maternal infection with HIV are associated with altered cardiovascular structure and function in utero. METHODS: Fetal echocardiography was performed in 173 fetuses of 169 HIV-infected mothers (mean gestational age, 33.0 weeks; SD = 3.7 weeks) at 5 centers. Biparietal diameter, femur length, cardiovascular dimensions, and Doppler velocities through atrioventricular and semilunar valves and the umbilical artery were measured. Measurements were converted to z scores based on published normal data. RESULTS: Fetuses determined after birth to be HIV-infected had similar echocardiographic findings as fetuses later determined to be HIV-uninfected except for slightly smaller left ventricular diastolic dimensions (P =.01). The femur length (P =.03) was also smaller in the fetuses postnatally identified as HIV-infected. Differences in cardiovascular dimensions and Doppler velocities were identified between fetuses of HIV-infected women and previously published normal fetal data. The reason for the differences may be a result of maternal HIV infection, maternal risk factors, or selection bias in the external control data. CONCLUSIONS: Vertically transmitted HIV infection may be associated with reduced left ventricular size but not with altered cardiac function in utero. Fetuses of HIV-infected mothers may have abnormal cardiovascular structure and function and increased placental vascular resistance, regardless of whether the fetuses are subsequently found to be infected with HIV. PMID- 11011331 TI - Misrepresentation of left ventricular contractile function by endocardial indexes: clinical implications after coarctation repair. AB - BACKGROUND: Endocardial function indexes overestimate myocardial fiber shortening, a geometric effect proportional to wall thickness. We hypothesized that elevated endocardial indexes of left ventricular contractile function after repair of isolated coarctation of the aorta could be related to this effect. METHODS: Chamber dimensions and wall thickness were measured from 59 echocardiograms in 57 patients aged 1.2 to 32 years, 8.5 +/- 5.6 years after coarctation repair, and in 305 normal controls aged 1 to 35 years. Midwall and endocardial shortening indexes and end-systolic fiber stress were calculated. The stress-velocity index (SVI), a load-independent index of contractility, was derived from these variables. All values were expressed as z scores. RESULTS: After coarctation repair, the midwall-derived SVI was elevated, but significantly less so than the endocardial-derived SVI (0.6 +/- 1.6 vs 1.3 +/- 2.6; P =.01). The endocardial-derived SVI correlated with the end-systolic thickness/dimension ratio (P <.0001), but the midwall-derived SVI did not. There was no linear relation between the midwall-derived SVI and the residual blood pressure gradient. The mean midwall-derived SVI was higher compared with the normal population in those with a minor residual blood pressure gradient (15 mm Hg), but this achieved statistical significance only in the latter group (0.5 +/- 1.6, P =.08; and 0.8 +/- 1.7, P =.03, respectively). CONCLUSIONS: Endocardial indexes of function and contractility overestimate fiber shortening after coarctation repair. Nevertheless, midwall shortening indexes demonstrate enhanced contractility, particularly in those with residual coarctation. PMID- 11011332 TI - Evaluation of left ventricular volumes and ejection fraction with a nonfluoroscopic endoventricular three-dimensional mapping technique. AB - BACKGROUND: Recently, a novel nonfluoroscopic 3-dimensional electromechanical mapping technique was introduced in the clinical arena. Although initial in vitro and in vivo studies suggested the reliability of the system in volumetric and hemodynamic evaluation of the left ventricle, no validation in human beings has been performed. METHODS: A nonfluoroscopic electromechanical mapping (NOGA, Biosense-Webster) procedure was performed in 44 patients. All patients received a contrast left ventriculogram during the same session. Volumetric (end-diastolic [EDV] and end-systolic volumes [ESV]) and hemodynamic (left ventricular ejection fraction [LVEF] and stroke volume) parameters of both systems were compared. RESULTS: Two uncomplicated pericardial effusions occurred with the first generation mapping catheters. No procedural complications were noted with the new generation mapping catheters. Significant correlations were found between mapping derived and ventriculography-based measurements for both ESV (r = 0.67, P <.001) and LVEF (r = 0.78, P <.001). Absolute volumes, however, were only comparable for ESV (46.6 +/- 25.3 mL vs 48.8 +/- 37.0 mL, respectively; P =.13) but differed greatly for LVEF (35% +/- 13% vs 65% +/- 19%, respectively; P <.001), EDV (69.1 +/- 28.6 mL vs 125.9 +/- 53.4 mL, respectively; P <.001) and stroke volume (22.4 +/- 9.9 mL vs 77.1 +/- 33.7 respirations; P <.001). Moreover, Bland-Altman analysis showed the clinical noninterchangeability between these techniques for the measurement of hemodynamic parameters. CONCLUSION: Measurement of hemodynamic parameters with nonfluoroscopic mapping of the left ventricle is feasible and safe. The system provides data that strongly correlate but that are in clinical disagreement with angiographic data. Therefore the interchangeability of these techniques may be questioned. PMID- 11011333 TI - Abciximab provides cost-effective survival advantage in high-volume interventional practice. AB - BACKGROUND: Placebo-controlled randomized trials of platelet glycoprotein (GP) IIb/IIIa blockade during percutaneous coronary intervention have demonstrated efficacy of these agents for reducing the risk of periprocedural ischemic events. However, cost-effectiveness of this adjunctive pharmacotherapy has been scrutinized. Extrapolation of cost-efficacy observations from clinical trials to "real world" interventional practice is problematic. METHODS: Consecutive percutaneous coronary interventions (n = 1472) performed by Ohio Heart Health Center operators at The Christ Hospital, Cincinnati, Ohio, in 1997 were analyzed for procedural and long-term (6-month) outcomes and charges. Observations on cost and efficacy (survival) were adjusted for nonrandomized abciximab allocation by means of "propensity scoring" methods. RESULTS: Abciximab therapy was associated with a survival advantage to 6 months after percutaneous coronary intervention. The average reduction in mortality rate at 6 months was 3.4% (unadjusted) and 4.9% when adjusted for nonrandomization. The average charge increment to 6 months was $1512 (unadjusted) and $950 when adjusted for nonrandomization. Patients deriving the greatest reduction in mortality rates also had a reduction in total cardiovascular charges to 6 months. Distinguishing demographics of this population included multivessel coronary intervention, coronary stent deployment, intervention within 1 week of myocardial infarction, and lower left ventricular ejection fraction. The average cost per life-year gained in this study was $2875 for all patients (unadjusted) and $1243 when adjusted for nonrandomization. CONCLUSIONS: Abciximab provides a cost-effective survival advantage in high volume interventional practice that compares favorably with currently accepted standards. Clinical and procedural demographics associated with increased cost effectiveness included multivessel coronary intervention, stent deployment, recent (<1 week) myocardial infarction, and impaired left ventricular function. PMID- 11011334 TI - Ventricular dilation after anterior ST-elevation myocardial infarction in the thrombolytic era. AB - BACKGROUND: The aim of this work was to study changes in end-diastolic volume 6 months after Q-wave and non-Q-wave anterior ST-elevation myocardial infarction by echocardiography. Ventricular dilation after anterior Q-wave myocardial infarction is well-recognized. However, there is a dearth of information about the natural history of ventricular volumes after non-Q-wave myocardial infarction. METHODS: One hundred ninety patients receiving thrombolytic therapy after anterior ST-elevation myocardial infarction were studied. All patients had 2D echocardiograms and 12-lead electrocardiograms recorded within 24 hours of symptoms and at 3, 42, and 180 days later. In addition, a further electrocardiogram was recorded on day 7 to assess patients for the presence of Q waves. Peak creatine kinase over the first 3 days of admission was recorded. End diastolic volume index was the study end point. RESULTS: Peak creatine kinase was strongly associated with ventricular dilation in both groups (P <.001). Mean end diastolic volume in the Q-wave group increased significantly from day 1 to 6 months (P <.05) but did not alter after non-Q-wave infarction. However, when patients were selected on predefined criteria for significant change in ventricular dilation (>10 mL/m(2)), then 35% of those with and 15% of those without Q waves fell into this category. Within this group, the increase in end diastolic volume followed a similar pattern, with the maximum percentage increase occurring between day 1 and 6 weeks. CONCLUSIONS: In the postthrombolytic group of anterior ST-elevation myocardial infarction, a minority of patients without Q wave development also undergo significant ventricular dilation. PMID- 11011335 TI - Renal endothelin in heart failure and its relation to sodium excretion. AB - BACKGROUND: Recent studies have shown that endothelin-1 (ET-1) antagonists increase sodium excretion and improve renal blood flow in experimental heart failure (HF). However, despite a number of investigations that have reported a significant increase in ET-1 plasma levels in patients with HF, it is still not known whether increased renal synthesis and urinary excretion of ET-1 occur. Our aim was to investigate renal ET-1 formation and its relation to sodium excretion in patients with HF. METHODS: One hundred forty-seven patients with HF, subdivided according to New York Heart Association (NYHA) functional classes, and 28 healthy controls were studied. ET-1 and big ET-1 were measured in plasma and in 24-hour urine by radioimmunoassay. Atrial and brain natriuretic peptide, arginine vasopressin, plasma renin activity, and hemodynamic variables were also investigated. RESULTS: Urinary ET-1 excretion was already increased in NYHA class II patients (P <.001 vs controls), whereas plasma ET-1 increased only in NYHA class III and IV patients (P <.001). In the 71 subjects who were not receiving diuretic treatment, urinary ET-1 was selected as the strongest predictor of sodium excretion by multivariate stepwise analysis. CONCLUSIONS: Urinary ET-1 excretion increases in an earlier phase of HF than plasma ET-1 and appears to be closely correlated with sodium excretion, indicating renal ET-1 is a target for ET-1 antagonists in patients with HF. PMID- 11011336 TI - Efficacy, safety, and effects on quality of life of bisoprolol/hydrochlorothiazide versus amlodipine in elderly patients with systolic hypertension. AB - BACKGROUND: Several studies have shown the benefits of antihypertensive treatment in elderly patients in terms of cardiovascular morbidity and mortality rate reduction. Low-dose drug combinations may be of interest in treating older subjects. A randomized, multicenter, double-blind, parallel group study was conducted to compare the efficacy and safety of bisoprolol 2.5 mg/hydrochlorothiazide 6.25 mg (n = 84) to amlodipine 5 mg (n = 80) in isolated systolic hypertension in patients older than 60 years. METHODS: After a 2- to 4 week placebo washout period, both drugs were administered once daily and taken for 12 weeks. Blood pressure was measured 24 hours after treatment administration. RESULTS: Systolic and diastolic blood pressure changes from baseline to week 12 were similar for both the bisoprolol and amlodipine groups ( 20. 0/-4.5 mm Hg and -19.6/-2.4 mm Hg, respectively). Overall adverse events for bisoprolol and amlodipine were 39% and 40%, respectively. Changes in quality of life scores were +2.5 for bisoprolol and +3.2 for amlodipine, with a positive change indicating improvement. CONCLUSIONS: This study demonstrates comparable efficacy and tolerability of bisoprolol 2.5 mg/hydrochlorothiazide 6.25 mg and amlodipine 5 mg. The low-dose combination of bisoprolol and hydrochlorothiazide may be an appropriate alternative for elderly patients with systolic hypertension. PMID- 11011337 TI - Long-term central hemodynamic effects at rest and during exercise of losartan in essential hypertension. AB - BACKGROUND: Losartan reduces blood pressure in patients with essential hypertension, but the long-term central hemodynamic effects at rest and during exercise are not known. METHODS AND RESULTS: After 8 months of losartan treatment (50 to 100 mg daily, mean 82 mg), intra-arterial pressure was reduced from 165/102 mm Hg to 145/91 mm Hg at rest and from 193/104 mm Hg to 179/96 mm Hg during 100 W exercise in 28 patients with essential hypertension. Cardiac index and heart rate remained unchanged, but total peripheral resistance index was reduced 12% to 15%. Stroke index was unchanged at rest but increased 7% to 9% during exercise. Twenty-four-hour ambulatory blood pressure was reduced 10% to 13%. Left ventricular mass was reduced 27% in patients with left ventricular hypertrophy (n = 18). CONCLUSION: Losartan lowers blood pressure by reducing total peripheral resistance at rest and during exercise but cardiac pump function is unchanged or slightly improved. In patients with left ventricular hypertrophy, losartan induces a sizeable reduction in left ventricular mass. PMID- 11011338 TI - Effect of niacin, warfarin, and antioxidant therapy on coagulation parameters in patients with peripheral arterial disease in the Arterial Disease Multiple Intervention Trial (ADMIT). AB - BACKGROUND: Patients with peripheral arterial disease (PAD) have high rates of cardiovascular morbidity and mortality, including that caused by associated coronary heart disease and cerebrovascular disease. Previous studies have shown that coagulation parameters are altered in PAD and that altered coagulation may play a critical role in the susceptibility to cardiovascular complications in PAD. It is therefore important to assess the effect of secondary prevention measures on coagulation in patients with PAD. The Arterial Disease Multiple Intervention Trial (ADMIT), a multicenter, randomized, placebo-controlled trial, was conducted to determine the feasibility of a combined lipid-modifying, antioxidant, and antithrombotic treatment regimen in patients with PAD. The objective of this study was to assess the effect of the ADMIT interventions on coagulation. METHODS: ADMIT participants were randomly assigned to low-dose warfarin, niacin, and antioxidant vitamin cocktail or corresponding placebos in a 2 x 2 x 2 factorial design. Specialized coagulation studies were performed in a subset of 80 ADMIT participants at baseline and after 12 months of treatment. RESULTS: Low-dose warfarin (1 to 4 mg/d) resulted in a significant decrease in factor VIIc (P <.001) and in plasma F1.2 (P =.001). Unexpectedly, niacin treatment also resulted in significant decrease in both fibrinogen (48 mg/dL; P <.001) and F1.2 (P =.04). von Willebrand factor increased after antioxidant vitamin treatment (P =.04). CONCLUSIONS: A regimen of low-dose warfarin effectively modifies coagulation in patients with PAD. Niacin also favorably modifies fibrinogen and plasma F1.2. Niacin, in addition to its lipid effects, modifies abnormal coagulation factors that accompany PAD. PMID- 11011339 TI - Superiority of enoxaparin versus unfractionated heparin for unstable angina/non-Q wave myocardial infarction regardless of activated partial thromboplastin time. AB - BACKGROUND: Whether the clinical superiority of enoxaparin versus unfractionated heparin (UFH) depends on a more stable antithrombotic effect or the proportion of patients not reaching the therapeutic level with UFH has not been addressed. METHODS: All patients participating in the Thrombolysis In Myocardial Infarction 11B trial who received UFH and had sufficient activated partial thromboplastin time (aPTT) data (n = 1893) were compared with patients who received enoxaparin (n = 1938). Patients receiving UFH were divided into 3 categories depending on mean aPTT values throughout 48 hours: subtherapeutic, for those in whom the average aPTT fell below 55 seconds; therapeutic, between 55 and 85 seconds; and supratherapeutic, longer than 85 seconds. Events and bleeding rates were determined at 48 hours. RESULTS: A small portion of patients (6. 7%) had a subtherapeutic average aPTT value (n = 127). Forty-seven percent of patients (n = 891) fell within the therapeutic range, and 46% were in the supratherapeutic level (n = 875). Event rates were 7. 0% in the UFH group versus 5.4% with enoxaparin (P =.039). Events rates were higher in every aPTT strata compared with enoxaparin and statistically significant in the supratherapeutic group (odds ratio 0.65; 95% confidence interval, 0.47-0.89). Major bleeding rates were 0%, 0.6%, and 0.9% for the subtherapeutic, target, and supratherapeutic strata, respectively, and 0.8% with enoxaparin. Minor hemorrhages occurred in 5.1% of patients receiving enoxaparin versus 3.9%, 2%, and 2.3%, respectively, for the UFH subgroups (P <. 001 for all UFH groups vs enoxaparin). CONCLUSIONS: Enoxaparin showed a better clinical profile compared with every level of anticoagulation with UFH. Potential mechanisms for enoxaparin superiority are stable antithrombotic activity, lack of rebound thrombosis, and intrinsic superiority. PMID- 11011340 TI - Effect of delay on racial differences in thrombolysis for acute myocardial infarction. AB - OBJECTIVE: To analyze the effect of delay times on racial differences in thrombolysis for acute myocardial infarction. BACKGROUND: Lower rates of thrombolytic therapy in blacks with acute myocardial infarction have recently been reported, but the reasons for this disparity are unknown. We hypothesized that lower rates of thrombolysis are caused by delay in presentation after symptom onset. METHODS: From November 1992 through November 1996, consecutive patients with a first acute myocardial infarction presenting to a large, urban teaching hospital were prospectively enrolled. Delay times were determined retrospectively from review of medical records. Patients were prospectively followed up for in-hospital cardiac events and death. A multivariable regression model was built to relate presentation times and other variables to thrombolysis administration. RESULTS: A total of 395 patients were included in the study, of which 33% were black. Symptom onset to emergency department presentation and door to-needle times were significantly longer in blacks. Thrombolysis was administered significantly less often in blacks compared with whites (47% vs 68%, P =.001). Black race and age above 60 years were independently associated with delayed presentation and prolonged door-to-needle times. Black race, time to presentation, and non-Q-wave myocardial infarction were independently associated with not receiving thrombolysis. In-hospital mortality rates were similar in both groups. CONCLUSIONS: Blacks presented later than whites for first acute myocardial infarction. Late arrival strongly influenced the rate of thrombolysis administration. Lower rates of thrombolysis and prolonged door-to-needle times were apparent in blacks after adjustment for delay times and other clinical factors, a finding that merits further investigation. PMID- 11011341 TI - Adequacy of intracoronary versus intravenous adenosine-induced maximal coronary hyperemia for fractional flow reserve measurements. AB - BACKGROUND: Fractional flow reserve (FFR) is a measure of coronary stenosis severity that is based on pressure measurements obtained at maximal hyperemia. The most widely used pharmacologic stimulus for maximal coronary hyperemia is adenosine, administered either as a continuous intravenous (IV) infusion or intracoronary (IC) bolus. IV adenosine has more side effects and is more costly than IC adenosine but has a more stable and prolonged hyperemic effect. METHODS: We compared the efficacy of IC and IV adenosine administration for the measurement of FFR in a multicenter trial. Fifty-two patients with 60 lesions underwent determination of FFR with both IV and IC adenosine. IV adenosine was administered as a continuous infusion at a rate of 140 microgram/kg per minute until a steady state hyperemia was achieved. IC adenosine boluses were administered at a dose of 15 to 20 microgram in the right and 18 to 24 microgram in the left coronary artery. FFR was calculated as the ratio of the distal coronary pressure (from pressure guide wire) to the aortic pressure (guide catheter) at maximal hyperemia. RESULTS: A total of 26 left anterior descending, 23 right, 9 left circumflex, and 3 left main coronary arteries were evaluated. Mean percent stenosis for both groups was 55.8% +/- 23.6% (range 0% to 95%), and mean FFR was 0.78 +/- 0.15 (range 0.41 to 0.98). There was a strong and linear correlation between FFR measurements with IV and IC adenosine (R = 0.978, y = 0. 032 + 0.964x, P <.001). The agreement between the 2 sets of measurements was also high, with a mean difference in FFR of -0.004 +/- 0.03. However, a small random scatter in both directions of FFR measurements was noted with 5 lesions (8.3%) where FFR with IC adenosine was higher by 0.05 or more compared with IV infusions, suggesting a suboptimal hyperemic response in these patients. Changes in heart rate and blood pressure were significantly higher with IV adenosine. Two patients with IV, but none with IC adenosine, had severe side effects (bronchospasm and severe nausea). CONCLUSION: These results suggest that IC adenosine is equivalent to IV infusion for the determination of FFR in the majority of patients. However, in a small percentage of cases, coronary hyperemia was suboptimal with IC adenosine. PMID- 11011343 TI - Changes in rates of beta-blocker use between 1994 and 1997 among elderly survivors of acute myocardial infarction. AB - BACKGROUND: Results of recent studies suggest that beta-blockers are underused by elderly acute myocardial infarction (AMI) survivors. The goals of this study were to examine changes in post-AMI beta-blocker use occurring between 1994 and 1997 and to identify factors associated with outpatient use of beta-blockers. METHODS: Patients included 9534 individuals >/=65 years of age who were enrolled in Pennsylvania's Pharmaceutical Assistance Contract for the Elderly (PACE) and who survived AMI between 1994 and 1997. With the use of outpatient prescription claims, beta-blocker usage rates were examined by year, and multivariate logistic regression was used to identify predictors of beta-blocker use. RESULTS: Post-AMI beta-blocker use increased from 39.6% in 1994 to 58.6% in 1997. Controlling for AMI year, individuals who had any prescriptions written by a cardiologist were more likely to use a beta-blocker than individuals who received all prescriptions from noncardiologists (odds ratio 1.52, P =.0001). Elderly patients who did not use beta-blockers tended to have greater severity of illness, non-Q-wave infarctions, atrial fibrillation, and comorbidities such as congestive heart failure, chronic obstructive pulmonary disease, and asthma. Use of calcium channel blockers and diuretics was negatively associated with beta-blocker use, but persons using lipid-lowering agents were more likely to use beta-blockers. CONCLUSIONS: Significant improvements in beta-blocker use by elderly AMI survivors have occurred since 1994. Differences among physician specialties in beta-blocker prescribing appear to remain. Across all prescriber specialties, historic contraindications constitute major factors in the choice of post-AMI therapy. Further efforts should be made to encourage the use of beta-blockers in elderly survivors of AMI, particularly those with heart failure. PMID- 11011342 TI - Incidence of left atrial thrombi in patients in sinus rhythm and with a recent neurologic deficit. AB - OBJECTIVES: We sought to determine the incidence of left atrial (LA) thrombi in patients in sinus rhythm (SR) and with a recent neurologic deficit and to analyze the relation between LA thrombi and LA chamber and appendage function in patients in SR. METHODS: A prospective study was conducted in 869 consecutive patients. The study group consisted of 583 patients in SR (67%). The remaining 286 patients had atrial fibrillation (AF) and served as controls (33%). RESULTS: The incidence of LA thrombi was significantly higher in patients with AF (n = 39 [14%]) compared with patients in SR (n = 6 [1%]; P <.001). Three of 6 patients with thrombi in SR had mitral stenosis, 1 patient had aortic stenosis, 1 patient had coronary artery disease, and another patient had a cardiomyopathy. Of the patients with detected thrombi, those in SR did not receive anticoagulation, whereas those with AF did in 18 cases. Patients with thrombi in SR and with AF did not significantly differ in LA diameter (5.1 +/- 0.8 cm vs 4.8 +/- 0.7 cm; 95% confidence interval [CI], -0.78 to 0.45), left ventricular ejection fraction (46% +/- 13% vs 42% +/- 15%; 95% CI, -18.7 to 7.4), LA appendage area (5.8 +/- 2.7 cm(2) vs 6.7 +/- 3.2 cm(2); 95% CI, -1.9 to 3.6), peak emptying velocity of the LA appendage (0.19 +/- 0.08 m/s vs 0.17 +/- 0.07 m/s; 95% CI, -0.08 to 0.04), or LA spontaneous echo contrast (3. 5 +/- 0.6 vs 3.9 +/- 0.5; 95% CI, -0.06 to 0.45). CONCLUSIONS: LA appendage thrombi are an infrequent cause of thromboembolism in patients in SR and are associated either with mitral valve disease or LA chamber and appendage dysfunction. Routine transesophageal echocardiography for the exclusion of LA thrombi is not recommended in patients in SR without underlying heart disease and normal LA function as assessed by transthoracic echocardiography. PMID- 11011344 TI - Twenty-four-hour autonomic nervous function in sustained and "white coat" hypertension. AB - BACKGROUND: It is unknown whether "white coat" hypertension, also known as isolated clinic hypertension, shares similarities in pathophysiologic background with sustained hypertension. Therefore we evaluated 24-hour autonomic nervous function in sustained and white coat hypertension. METHODS: We selected 12 patients with sustained hypertension (clinic blood pressure >/=140/90 mm Hg and daytime blood pressure >135/85 mm Hg) and 12 patients with white coat hypertension (clinic blood pressure >/=140/90 mm Hg and daytime blood pressure <135/85 mm Hg) from patients undergoing ambulatory blood pressure monitoring and 12 normotensives for study inclusion. Groups were matched for age, sex, and body mass index and had similar dietary pattern and occupational status (civil servants with sedentary jobs). Subjects underwent noninvasive 24-hour monitoring of blood pressure, R-R interval of the electrocardiogram, body position, activity rate, and ambient temperature. Power spectral analysis of R-R intervals was performed with an autoregressive model to obtain the low-frequency component, the high-frequency component, and their ratio. Subjects also collected 24-hour urine samples for examination of norepinephrine and epinephrine excretion by high performance liquid chromatography. RESULTS: Work and sleep time, body position, ambient temperature, and activity were not different among the groups. Daytime, nighttime, and 24-hour low-frequency/high-frequency ratios were significantly higher in patients with sustained hypertension than in patients with white coat hypertension (3.4 +/- 0.45 vs 2.65 +/- 0.45, 2.35 +/- 0.60 vs 1. 82 +/- 0.45, and 3.04 +/- 0.45 vs 2.4 +/- 0.35, respectively, P <. 05). Urinary norepinephrine excretion (53 +/- 12 microg vs 29.5 +/- 6 microg; P <.05) and vanillylmandelic acid excretion (4.45 +/- 0.6 mg vs 3.1 +/- 0.55 mg; P <.05) during the 24 hours were significantly higher in patients with sustained hypertension than in those with white coat hypertension. There was no difference between those with white coat hypertension and normotensives concerning the aforementioned parameters. CONCLUSIONS: Our findings indicate whole-day sympathetic overactivity in sustained hypertension but not in white coat hypertension, suggesting that these conditions show some differences in pathophysiologic background. PMID- 11011345 TI - Sex differences on the electrocardiographic pattern of cardiac repolarization: possible role of testosterone. AB - BACKGROUND: Cardiac repolarization has been shown to be shorter and faster in men compared with women. In this study, we examined the electrocardiographic pattern of repolarization in patients with abnormal plasma levels of testosterone to gain insight into the role that this hormone plays in modulating repolarization. METHODS AND RESULTS: Repolarization was measured in 27 castrated men, 26 women with virilization, and 53 control subjects pair-matched for age and sex. Repolarization in castrated men was slower and longer than that of normal men. Women with virilization exhibited a shorter and faster repolarization than normal women and castrated men. These differences are the opposite of those found in the normal population. Finally, the changes observed in castrated men may be reverted by testosterone. CONCLUSIONS: We conclude that testosterone plays an important role in modulating cardiac repolarization. PMID- 11011346 TI - Endothelium-dependent responses in patients with hypercholesterolemic coronary artery disease under the effects of simvastatin and enalapril, either separately or combined. AB - BACKGROUND: Endothelial dysfunction is an early event in atherosclerotic disease that precedes clinical manifestations and complications. The noninvasive assessment of endothelial function in patients with risk factors undergoing clinical treatment is an important medical advance. In this setting, altered endothelial function in patients with coronary hypercholesterolemia and its modifications by treatment with enalapril and simvastatin, either separately or in combination, was assessed in the brachial artery in a randomized, double blind, 2-period crossover study. METHODS: Thirty-eight patients were separated in 2 groups. Group 1 (18 patients, 3 female, mean age 63 +/- 6.0 years) received simvastatin 10 mg/d for 8 weeks and simvastatin plus enalapril 5 mg/d for another 8 weeks. Group 2 (20 patients, 3 female, mean age 64 +/- 5.8 years) received enalapril 5 mg/d for 8 weeks and enalapril plus simvastatin 10 mg/d for another 8 weeks. All subjects underwent measurements of brachial artery diameter before and after postischemic hyperemia with high-resolution ultrasound at basal conditions (control) and under the effects of the drugs at the end of 8 and 16 weeks. RESULTS: Cholesterol and LDL cholesterol levels significantly decreased with simvastatin treatment alone or with enalapril. Mean baseline arterial diameter was 5.24 +/- 1.25 mm in group 1 and 4.83 +/- 0.99 mm in group 2 (not significant). In group 1 after the first 8 weeks, endothelium-dependent vasodilation significantly increased with simvastatin treatment (control, 4.4%; 8 weeks, 7.6%; P <.001). After 16 weeks with the addition of enalapril, a further increase in vasodilation was seen (8.6%, P <.05 vs 8 weeks). In group 2, with enalapril treatment an increase in vasodilation versus control was seen (control, 4.3%; 8 weeks, 5.8%; P <.01). After 16 weeks, with the addition of simvastatin an additional increase in vasodilation was observed (9.1%, P <.001 vs 8 weeks). After nitroglycerin, vasodilation in group 1 at control, 8, and 16 weeks was 17%, 17.5%, and 18%, respectively. In group 2, nitroglycerin vasodilation at control, 8, and 16 weeks was 21%, 21%, and 22%, respectively. CONCLUSIONS: Simvastatin significantly increased the postischemic vasodilator response in patients with coronary hypercholesterolemia, either as single treatment or added to enalapril. Similarly, the response was increased by enalapril, either alone or while simvastatin was being administered. Both drugs improve vasodilation and additive effects appear to be present. PMID- 11011347 TI - Cardiac biochemical markers after cardioversion of atrial fibrillation or atrial flutter. AB - BACKGROUND: Cardioversion or defibrillation of cardiac arrhythmias is often necessary in acutely ill cardiac patients. The electrical current may cause elevation of biochemical markers used to diagnose acute myocardial infarction. Therefore it is important to find cardiac markers with high specificity for myocardial necrosis. The purpose of this study was to assess the effects of elective cardioversion of atrial fibrillation or flutter on troponin T and I among conventional markers in patients with no evidence of acute ischemia. METHODS AND RESULTS: Fifty-seven consecutive patients underwent 1 to 4 direct current shocks (mean cumulative energy 407 J, range 100 to 920 J) under general anesthesia. At baseline, all had normal troponin levels; 50 patients (mean age 68 years, range 33 to 84 years) had normal cardiac enzymes and were included in the final analysis. Blood samples were drawn at baseline, and 1 to 2, 6 to 8, and 20 to 24 hours after cardioversion. The troponin levels were unaffected by cardioversion in all patients, whereas creatine kinase and myoglobin increased more than 10-fold. Creatine kinase MB mass and aspartate aminotransferase were above reference limits in 18% and 24% of patients, respectively, 20 to 24 hours after cardioversion. There was a significant association between elevated creatine kinase, myoglobin, and creatine kinase MB levels with cumulated energy delivered as well as when possible confounders such as age and sex were adjusted for. High international normalized ratio with warfarin use was associated with increased levels of creatine kinase, aspartate aminotransferase, lactate dehydrogenase, and myoglobin. CONCLUSIONS: The increase of conventional biochemical markers after direct current cardioversion is positively associated with cumulative energy delivered and international normalized ratio (INR) values; neither influences levels of the cardiac troponins. PMID- 11011348 TI - Introduction. PMID- 11011350 TI - Depression and ischemic heart disease. AB - Major depression is a common comorbidity associated with ischemic heart disease (IHD). There is growing evidence that psychological stress in general and depression in particular predispose to cardiovascular disease. Persons who have mental stress during daily life are at twice the risk of myocardial ischemia, and patients with post-myocardial infarction depression have higher mortality rates than nondepressed controls. These data suggest a psychophysiologic mechanism underlying the vulnerability of depressed patients to IHD. Clinical studies have demonstrated that depression is associated with a much higher risk of both cardiovascular morbidity and mortality, which could be caused by platelet activation. Physicians should maintain a heightened level of clinical suspicion for depression and depressive disorders in persons with IHD, particularly those individuals who are recovering from an acute ischemic event, such as myocardial infarction. Furthermore, depression may complicate the recovery of IHD, but in most cases depression can be effectively treated with antidepressant agents. PMID- 11011349 TI - Are platelets the link between depression and ischemic heart disease? AB - The role of platelets as the link between ischemic heart disease and depression is reviewed. Platelet function abnormalities, including increased platelet reactivity, may predispose depressed patients to clotting diatheses and may explain their vulnerability to cardiovascular disease. Platelet physiologic characteristics, influences on platelet response, and indicators of platelet reactivity are discussed. Measurements of platelet activation, secretion, and aggregation have enabled the study of platelet responses in patients with major depression. The findings of clinical trials evaluating platelet responses to antidepressant treatment are described, and recommendations for future studies are proposed. PMID- 11011351 TI - Depression as a contributing factor in cerebrovascular disease. AB - Stroke is a leading cause of morbidity and mortality in the United States. Physical and psychological impairment from stroke may negatively affect quality of life. From a psychological perspective, depression, which is common in stroke patients, may influence functional recovery and possibly mortality after a stroke. Although somewhat controversial, the risk of poststroke depression may be related to lesion location, specifically left anterior or left basal ganglia lesions. Because poststroke depression can affect recovery, treatment with antidepressants may be warranted. Emerging evidence suggests that nortriptyline and fluoxetine may be effective; paroxetine, sertraline, and other selective serotonin reuptake inhibitors have shown efficacy in treating a related phenomenon known as pathologic crying. The influence of depression on the risk of stroke has not been extensively studied; however, several epidemiologic studies raise the possibility that a relation exists. In one study of elderly individuals, higher depressive symptoms increased the risk of stroke. Many older individuals who have depression develop later in life have evidence of subtle cerebrovascular disease. Specifically, silent strokes that do not involve the sensory or motor parts of the brain are quite common in the elderly population. Vascular depression is depression that occurs in patients with cerebrovascular or ischemic changes in the brain. Preliminary evidence indicates that these vascular changes may be related to atherosclerosis, hypertension, or myocardial infarction. However, additional studies are needed to gain a better profile of these patients and to explore potential treatment modalities. PMID- 11011352 TI - Heart rate variability in depressive and anxiety disorders. AB - Loss of normal autonomic nervous system control of heart rate and rhythm is an important risk factor for adverse cardiovascular events. After myocardial infarction, reduction in beat-to-beat heart rate variability, a measure of cardiac autonomic innervation by the brain, is a strong predictor of death. With loss of vagal innervation, as is noted in patients with severe neuropathy and in heart transplant recipients, there is loss of heart rate variability. It is speculated that decreased parasympathetic innervation exposes the heart to unopposed stimulation by sympathetic nerves. Individuals with high hostility scores and patients with anxiety or depressive disorders have low heart rate variability and may be at increased risk for cardiovascular death associated with coronary heart disease and arrhythmias. After myocardial infarction, depressed patients exhibit higher mortality rates compared with nondepressed patients. Men with "phobic anxiety," a construct that appears to overlap substantially with panic disorder, also have higher rates of sudden cardiac death and coronary artery disease than control populations. The reduction in autonomic nervous system control to the heart may be one link between psychopathology and heart disease. Although tricyclic antidepressants reduce heart rate variability, at least one study has suggested that, in patients with panic disorder, treatment with the selective serotonin reuptake inhibitor paroxetine normalizes heart rate variability. Hence there is potential for the treatment of psychiatric disorders to affect positively the development and course of cardiovascular disease. PMID- 11011353 TI - Considerations for the use of antidepressants in patients with cardiovascular disease. AB - There is convincing evidence that depression can significantly and adversely affect cardiovascular health and increase mortality rates in patients with documented ischemic heart disease. It is unknown whether treatment of depression can reduce the risk of IHD or if treatment can decrease mortality rates after myocardial infarction. Nonetheless, the available evidence strongly suggests that depression in patients with cardiovascular disease should be treated. Tricyclic antidepressants had been considered acceptable for use in patients with ischemic heart disease until data from the Cardiac Arrhythmia Suppression Trial (CAST) demonstrated a significantly increased mortality rate after myocardial infarction in patients treated with type I antiarrhythmics. Because tricyclic antidepressants are type IA antiarrhythmics, they presumably carry a risk similar to that of moricizine in patients with ischemic disease. The limited but growing data available on the use of selective serotonin reuptake inhibitors and bupropion in patients with cardiac disorders suggest that these agents are safer antidepressant treatment alternatives. Larger, long-term, randomized, controlled studies are needed to confirm that selective serotonin reuptake inhibitors are indeed safe in depressed patients with cardiovascular disease. PMID- 11011363 TI - [Topical organization of the cat inferior colliculus under conditions of the simulated sound source motion]. AB - Binaurally presented click trains with gradually changing ITDs were used to imitate the sound source motion. Focal EP-trains from the cat inferior colliculus, IC, were recorded, and changing amplitude of the successive EPs following successive clicks was taken for estimation of the motion effect (ME). It was shown that a) in 25% of cases the ME depends on the motion direction; b) the ME is better presented ipsilaterally in respect to the recording point, with equal dependence on motion velocity both on the left and on the right; c) the ME phenomenon is connected with prevalence of contralateral afferentation as compared to the ipsilateral one; d) the ME is connected with inhibitory phenomena such as binaural suppression of monaural afferentation; e) the ME of the best amplitude is most pronounced in the ventrolateral area of the IC central nucleus, its localisation being near to that of the great multipolar neurones, whereas slightly and moderately pronounced MEs are presented rather uniformly over the central nucleus like the "basic" neurones are; MEs of different pronouncement are supposed to depend differently on effectiveness of ipsi- and contralateral stimulation--in accordance with properties of multipolar and "basic" neurons. PMID- 11011362 TI - [The role of ventrolateral thalamic nucleus in switching of the descending influence over to the rat motor activity]. AB - Facilitated influence of preliminary transection of the rat rubrospinal tract on motor activity and instrumental reflexes recovering after lesion of the red nucleus was more obvious after a chemical lesion rather than the electrolytic lesion. This seems to be due to remaining cerebello-thalamic fibres after a chemical lesion of the red nucleus. A preliminary destruction of the ventrolateral thalamic nucleus was shown to complicate switching of the motor activity in the rats with transected rubrospinal tract and lesioned red nucleus. PMID- 11011364 TI - [Comparison of binaural release from forward masking in animals and humans. Electrophysiologic study]. AB - The EPs of the inferior colliculus and auditory cortex in anaesthetized guinea pigs and the long latency auditory EPs in alert humans were studied. The stimuli consisted of binaurally presented pairs of clicks used as a masker, and the probe, with a variable time delay between them. The greatest relative differences between out-of-phase and in-phase probe responses were observed at the beginning of the recovery course. They averaged as 1.6, 1.5 and 1.4 for the responses of the inferior colliculus, auditory cortex and long latency potentials, resp., at the stimuli intensities of 50-65 dB SPL, and then decreased to zero during the time course of the probe response recovery. Correlation of this parameter with the stimulus intensity was positive. PMID- 11011365 TI - [Mechanisms of the otolith signal processing]. AB - A series of current hypotheses of mechanisms of the otolith signals processing and organization of adaptive responses of the oculomotor system to gravito sensitive (otolith) inputs are reviewed: the "frequency segregation of the otolith signals", "gravito-inertial force resolution", "spatio-temporal transformation of the angular vestibulo-ocular reflex", "inertial processing of the vestibular signals", "asymmetry of the otolith signals", and their experimental corroboration. PMID- 11011368 TI - [Auditory analysis of the sound source approaching and withdrawing: psychoacoustic and neurophysiological correlates]. AB - The findings seemed to be based on direction and velocity of modelling the radial sound source shifting in free acoustic field. The threshold and the optimal parameters of the acoustic model imitating the approaching and withdrawing of the sound source shifting in silence and under conditions of noise, were established. A correlation between peak-to-peak amplitudes of the N1-P2 components of auditory EPs and the imitated direction of the sound shifting, was shown. The role of different left and right hemispheres' areas in perception of the radial sound source was analysed. The detector features of the central auditory neurones were shown as a possible mechanism of estimating the sound source approaching and withdrawal. PMID- 11011367 TI - [Disinhibition as a mechanism of tuning of cross-like figures in the visual cortex neurons]. AB - A discrete neural net was used for simulation of cross-sensitivity in 40% of neurones of the cat visual cortex' area 17th. It is based on disinhibition of the end-stopping inhibition in receptive field from the side-disinhibitory zone. Highly selective or invariant sensitivity of the simulated neurone in respect to shape and orientation of a cross-like figure was observed under changes of location, size and weight of the receptive field zones. The disinhibitory mechanism seems to be critically involved in the selection of the second-order features of the images in the primary visual cortex. PMID- 11011366 TI - [Intercellular integration in the central nervous system]. AB - A humoral mechanism of the intercellular communication in the CNS based on diffusion of neuroactive compounds within the brain extracellular space, was studied. The leading role of the volume transmission was shown in the early stage of ontogeny. Structural basis of this mechanism was studied in adult mammals, and the data on extrasynaptic receptors and release of classical neurotransmitters into the extracellular space was reviewed. PMID- 11011369 TI - [Effect of serotonin on isolated cells with the various functionality from the lamprey spinal cord]. AB - The differential actions of 5-hydroxytryptamine (5-HT) (100 microM) were investigated on isolated motoneurons, interneurons, and primary sensory neurons from the lamprey spinal cord using patch-clamp techniques. Application of 5-HT did not evoke membrane currents in any of the spinal neurons tested (n = 62). However, in most motoneurons and interneurons (15 of 18), 5-HT produced a small depolarization (2-6 mV), which was not accompanied by a change in input resistance. In the remaining motoneurons and interneurons (3 of 18), 5-HT induced a large depolarization (up to 10-20 mV) and a decrease in input resistance of 20 60%. In most sensory neurons (dorsal sensory cells, DSCs), 5-HT evoked a short lasting, low-amplitude depolarization, followed by a long-lasting hyperpolarization of 2-7 mV. The DSCs showed no significant change in input resistance to 5-HT application (n = 8). Spike afterpolarization were also differentially modulated by 5-HT. In motoneurons and interneurons, 5-HT decreased the amplitude of the afterhyperpolarization following the action potential while increasing the amplitude of the after depolarization. In the DSCs, no significant effect of 5-HT on spike afterpolarization was observed. 5-HT differentially modulated the current induced by application of N-methyl-D-aspartate (NMDA). In motoneurons and interneurons, 5-HT enhanced NMDA-evoked current, while in DSCs, 5 HT decreased this current. These results demonstrate that 5-HT differentially modulates the activity of functionally different groups of spinal neurons. In motoneurons and interneurons, 5-HT enhances excitation by inducing depolarization and decreasing the afterhyperpolatization, while NMDA currents are enhanced. These effects facilitate the appearance of rhythmic discharges in these cells in the presence of NMDA. In primary dorsal sensory cells, 5-HT enhances inhibition by hyperpolarizing the cells and depressing NMDA currents. These differential effects are presumably mediated by different types of 5-HT receptors on these classes of spinal neurons. PMID- 11011370 TI - [Studies of directional sensitivity of neurons in the cat primary auditory cortex]. AB - A set of impulsive transient signals has been synthesized for earphone delivery whose waveform and amplitude spectra, measured at the eardrum, mimic those of sounds arriving from a free-field source. The complete stimulus set forms a "virtual acoustic space" (VAS) for the cat. VAS stimuli are delivered via calibrated earphones sealed into the external meatus in cats under barbiturate anesthesia. Neurons recorded extracellularly in primary (AI) auditory cortex exhibit sensitivity to the direction of sound in VAS. The aggregation of effective sound directions forms a virtual space receptive field (VSRF). At about 20 dB above minimal threshold, VSRFs recorded in otherwise quiet and anechoic space fall into categories based on spatial dimension and location. The size, shape and location of VSRFs remain stable over many hours of recording and are found to be shaped by excitatory and inhibitory interactions of activity arriving from the two ears. Within the VSRF response latency and strength vary systematically with stimulus direction. In an ensemble of such neurons these functional gradients provide information about stimulus direction, which closely accounts for a human listener's spatial acuity. Raising stimulus intensity, introducing continuous background noise or presenting a conditioning stimulus all influence the extent of the VSRF but leave intact the gradient structure of the field. These and other findings suggest that such functional gradients in VSRFs of ensembles of AI neurons are instrumental in coding sound direction and robust enough to overcome interference from competing environmental sounds. PMID- 11011371 TI - [Visual, auditory, and bimodal activity in the banks of the lateral suprasylvian sulcus in the cat]. AB - In addition to visually driven cells we found within the lateral suprasylvian visual cortex of cats a considerable number of auditory and/or bimodal cells. Most of the visually driven cells were direction and orientation selective with responses that were neither highly stimulus time locked nor very stable. Most of the auditory responses were also not very stable, had relatively high thresholds and were readily habituated. Previous studies have suggested that populations of cells within the lateral suprasylvian area are specialized for the analysis of optic flow fields. Given that a remarkable proportion of cells within this area can be also driven by auditory stimuli we hypothesize that the "optic flow" model may be extended to the bimodal domain rather than restricted to visual clues only. This, however, remains to be corroborated experimentally. PMID- 11011372 TI - [Neural mechanisms of sound localization in owls]. AB - Barn owls localize sound by using the interaural time difference of the horizontal plane and the interaural intensity difference for the vertical plane. The owl's auditory system possesses the two binaural cues in separate pathways in the brainstem. Owls use a process similar to cross-correlation to derive interaural time differences. Convergence of different frequency bands in the inferior colliculus solves the problems of phase-ambiguity which is inherent in cross-correlating periodic signals. The two pathways converge in the external nucleus of the inferior colliculus to give rise to neurons that are selective for combinations of the two cues. These neurons form a map of auditory space. The map projects to the optic tectum to form a bimodal map which, in turn, projects to a motor map for head turning. The visual system calibrates the auditory space map during ontogeny in which acoustic variables change. In addition to this tectal pathway, the forebrain can also control the sound-localizing behaviour. PMID- 11011373 TI - Killer waves on the East coast? PMID- 11011374 TI - The roots of homicide. PMID- 11011376 TI - Biology--cell death. One-hit wonder. PMID- 11011375 TI - Womb wars. PMID- 11011377 TI - Speech without accountability. PMID- 11011378 TI - The wireless Web. PMID- 11011379 TI - The Internet in your hands. PMID- 11011380 TI - The promise and perils of WAP. PMID- 11011381 TI - The future is here. Or is it? PMID- 11011382 TI - The third-generation gap. PMID- 11011383 TI - Operating on a beating heart. PMID- 11011385 TI - Animals imitate, too. PMID- 11011384 TI - The power of memes. PMID- 11011386 TI - Meme theory oversimplifies how culture changes. PMID- 11011387 TI - People do more than imitate. PMID- 11011388 TI - Nabada: the buried city. PMID- 11011389 TI - Better decisions through science. PMID- 11011391 TI - Down among the micrograms. PMID- 11011390 TI - Contact lenses. The hard and the soft. PMID- 11011392 TI - Million-dollar minesweeper. PMID- 11011393 TI - Getting high. Booze, dope, metaphysics and other rhapsodic matters. PMID- 11011394 TI - [Nikolai Vladimirovich Ekk (1849-1908)]. PMID- 11011395 TI - ["Multiple organ failure syndrome". Nature of the concept and correctness of the nomenclature]. AB - The syndrome principle of studying complex pathological processes is justified if the key link of the pathogenesis is determined. The multi-organ systemic dysfunction is due to altered synergism of all visceral functions rather than to the sum of functional alterations of several organs. The main ideas and regularities of the synergism as the fundamental theoretical directions determine the coordination of vital activity and the probability of its disturbance depending on genetically determined and other factors. Grounds are presented for the prospects of using the Mandelbroth's fractals and regularities of synergism in creation of the prognostic dynamic models in the organism's extreme states. PMID- 11011397 TI - [Strategy of treatment of aortic stenosis complicated by total calcinosis]. AB - The authors made an analysis of problems of strategy of surgical treatment of 132 patients with aortal stenosis complicated by calcinosis. Detailed indications to surgical treatment for stenosis of the aortal valve are given depending on the stage of the disease and complications. The nuances of the technique of performing the operation of prosthesis of the aortic valve are described. PMID- 11011396 TI - [Age-dependent dynamics of biomechanical and hydrodynamic properties of the valve aortic complex of man]. AB - A comparative analysis of geometrical, biomechanical and hydrodynamic characteristics of the valve-aortic complexes taken from humans of different age has shown a determining role of distensibility of the aortic root in the functioning of valve cusps. An alteration of geometrical relationships of elements of the aortic root, its less distensibility, worse biomechanical and hydrodynamic characteristics of the aortic valve in humans older than 50-60 years of age is established. PMID- 11011398 TI - [A comparative evaluation of different methods of esophagoplasty]. AB - Intraoperative comparative oxyhemo- and thermometry of fragments of the gastrointestinal tract used for creation of the artificial esophagus were performed in 41 patients. The relationship of the degree of ischemic alterations in the transplant tissues and the development of postoperative complications was determined. PMID- 11011399 TI - [Long-term outcome of surgical treatment of patients with esophageal cancer]. AB - Under study were the results of surgical treatment of 185 patients with esophageal carcinoma. One-year survival was 75.4%, 2-year survival was 49.6%, 3 year--43.4%, and 4-year survival--33.3%. Among the patients with tumors with the invasion degree T1, T2, T3 and having no metastases to the regional lymph nodes the resection of the esophagus in combination with the extended lympho dissection resulted in 4-year survival in more than 50% of the patients operated upon. In patients with tumors T3N1, T4N1 and especially with the simultaneous involvement of several lymph collectors the 4-year survival was 21.4%. The operative intervention at this stage of the disease should be considered as a palliative means in the greater part of the patients. PMID- 11011400 TI - [Surgical treatment in cancer of the right half of the colon complicated by occlusive ileus]. AB - Results of treatment of 68 patients with carcinoma of the right half of the colon complicated by occlusive ileus are presented. The operation of choice is thought to be radical surgical intervention with the primary ablation of the tumor. Palliative operations were performed on 28 (41.2%) of 68 patients with occlusive ileus of the right half of the colon having risk factors, radical operations--on 40 patients (58.8%). Seven patients died at the postoperative period. Over-all lethality was 10.3%. PMID- 11011401 TI - [Mechanism of action of paravasal infiltration]. AB - In order to study the mechanisms of action of paravasal infiltration the content of middle mass molecules and general thiol groups in the central and regional blood flow were studied in 65 patients with obliterating atherosclerosis. The relationship between the stage of extremity ischemia and concentration of general thiol groups was shown. PMID- 11011402 TI - [Assessment of the value of certain clinical and laboratory indices in improvement of the diagnosis of acute appendicitis in children]. AB - The results of diagnosis and treatment of 1483 children were analyzed in order to determine the frequency of clinical symptoms and laboratory indices in children with acute appendicitis. Seven most valuable objective parameters were stressed which allowed to differentiate the strategy in situations difficult for diagnosis. PMID- 11011403 TI - [Surgical treatment of children with autosomal dominant polycystic kidney disease]. AB - The authors dealt with 59 children with different variants of polycystic kidney disease. The age of the patients was from 1 month to 19 years. The autosomal dominant type of inheritance was noted in 52 patients, in 24 of them percutaneous ignipuncture, lumbotomy with ignipuncture and dissection of the cysts, endovideosurgical opening and dissection of the cysts transperitoneally and retroperitoneally were performed. Long-term results were followed during the periods from 6 months till 17 years. Good results were obtained in 12 patients, satisfactory--in 2 patients, unsatisfactory--in 7. The most effective methods are traditional lumbotomy and endovideosurgical correction which are not followed by growth of cysts in the operated kidneys at the postoperative period. PMID- 11011404 TI - [Infectious complications in electric cardiac stimulation]. AB - There were 4424 patients operated for alterations of the cardiac rhythm. Primary electrocardiostimulation was performed in 3183 of them, reoperations in 1241 patients (39%). Infectious complications took place in 146 patients (3.3%). Based on their experiences the authors developed the strategy of treatment of patients with purulent complications and sepsis depending on their character and time of their appearance. PMID- 11011405 TI - [New trends in the treatment of liver abscesses]. AB - The authors share their experiences with treatment of 85 patients with abscesses of the liver aged from 17 to 83 years. The size of abscesses was from 2.5 to 23 cm in diameter. For the topical diagnostics the following methods were used: ultrasound scanning, computed tomography, scintigraphy, diagnostic puncture biopsy of the liver, laparoscopy etc. Decision between the methods of operative treatment depended on the concrete clinical situation. Both open and minimally invasive methods were used. Hydropressive technologies and ozone therapy were used for better sanitation of the abscess cavity. A new method of the assisted minimally invasive intervention was developed especially for treatment of patients with large abscesses of the liver. The new technologies allowed to reduce the amount of postoperative complications and to make the time of staying at the hospital substantially shorter. PMID- 11011406 TI - [Hemorheological indices in patients with amebic abscesses of the liver and their correction]. AB - Hemorheological indices were studied in 89 patients. In 61 patients the authors have revealed somewhat increased viscosity potential of blood and negligible changes in the aggregation activity of erythrocytes which is thought by the authors to be due to the saved compensatory potentialities of the organism. In 28 patients with the complicated course of the disease the rheological parameters of blood became worse because of the suppressed compensatory potentialities of organism. So, in addition to the adequate transfusion-infusion therapy the complex of treatment must include immunostimulating means (Levamisole, Prodigiozan, Tactivin etc.). PMID- 11011407 TI - [Antioxidant therapy with xymedon of patients with chronic osteomyelitis]. AB - An investigation of the antioxidant activity of blood plasma allowed a conclusion to be made of an alteration of the free radical processes in patients with chronic osteomyelitis. Experimental data on the influence of the antioxidants Xymedon and Methyluracil on the main free radicals, lipid peroxidation and chemiluminescence of peritoneal macrophages of rats allowed the substantiation of using the medicines in the complex treatment of patients in the pre- and postoperative periods. As a result, in 25 patients Xymedon improved the blood indices, increased antioxidant activity of plasma and reduced the amount of recurrences of the disease as compared with the patients taking Methyluracil. PMID- 11011408 TI - [Complex treatment of patients with complicated forms of erysipelas]. AB - The authors made an analysis of variants of treatment strategy and its results in patients with different forms of erysipelas. The use of complex conservative and operative treatment allowed lethality to be reduced from 4 to 1.9%, in patients with severe course of the disease the frequency of complications being reduced to 10%. PMID- 11011409 TI - [Extracorporeal hemocorrection in acute pancreatitis]. AB - An experience with using 340 operations of extracorporeal hemocorrection in complex intensive therapy of 160 patients with acute pancreatitis has been generalized. In 111 of these patients (69%) pancreatic necrosis complicated by the syndrome of multiple organ failure was diagnosed. Based on the mechanisms of medical efficiency the authors have developed differential indications for using different extracorporeal technologies depending on the clinico-laboratory profile of the endogenous intoxication, structure and degree of organic and systemic dysfunctions. The adoption of such technologies allowed lethality to be reduced from 37.5 to 27.6%. PMID- 11011410 TI - [Determination of indications for preoperative blood transfusion in patients with chronic ulcer of the stomach and duodenum complicated by hemorrhage]. AB - Results of the examination and treatment of 61 patients with chronic gastroduodenal ulcers complicated by bleeding have been analyzed. The possibility is shown to use hemoglobin indices for the estimation of severity of bleedings and for determination of indications for preoperative hemotransfusion to patients with chronic gastroduodenal ulcers complicated by bleeding. It was found that to refuse or reduce blood transfusion at the preoperative period was possible depending on the initial level of hemoglobin with the purpose of blood saving and in order to decrease the risk of reactions to hemotransfusion and complications. PMID- 11011412 TI - [Video endoscopy of the residual cavity in hydatidectomy of the liver]. AB - Results of treatment of 162 patients with echinococcosis of the liver were analyzed. Traditional surgical treatment of these patients was followed by endovideoscopy of the residual cavity of the liver. During endovideoscopy of the residual cavities the remnants of the chitinous membranes and small daughter vesicles not noticed during the open stage of the operation were removed in 7.4% of the patients, in 25.9% of the patients cystobiliary fistulas were detected. The endovideoscopy considerably improves revision of the residual cavities of the liver and facilitates liquidation of cystobiliary fistulas which gives better results of the treatment of such patients. PMID- 11011411 TI - [Dynamics of fluid volumes of body sectors in patients after laparoscopic surgery]. AB - An investigation of the state of volumes of the organism's fluid sectors (general, extra- and intracellular fluid) by a noninvasive method of double frequency impedancemetry has revealed the influence of preoperative preparing the patients and restriction of the water regimen after operation upon the development of moderate dehydration of the extracellular sector. During the operation lasting about an hour the intravenous administration of 400 ml of crystalloid solutions maintains the volumes of fluid sectors within the due values. These solutions injected in greater volumes result in accumulation of the extra- and intracellular fluid. The antiperistaltic purgation of the intestine before the operation reduces the concentration of calcium in blood by 1 mmol/l at an average. PMID- 11011413 TI - [Nephrectomy in general surgical hospital]. AB - In the central regional hospital 50 nephrectomies were performed for traumas and different diseases of the kidney, 44 of them being fulfilled through a laparoscopic access. This access allowed the volume of operation to be expanded in 58% of the patients. In 8% of the patients massive hemorrhage took place. Four patients died. Nephrectomy following pretreatment of the renal vessels allows to reduce risk of the operation. PMID- 11011414 TI - [Surgical treatment of middle-aged and elderly patients with chronic ischemia and distal occlusions]. AB - Operative treatment for critical ischemia was performed on 346 elderly and senile patients. An assessment of the distal bed and outflow pathways has shown the conditions for the reconstruction of the vessels to be unfavorable in 257 (74%) patients. Femoro-proximal-popliteal shunts were made in 82 patients with good effect in 74 of them. Femoro-distal-popliteal and tibial shunts were made in 62 patients with good effect in 52. In 12 patients out of 20 with the isolated plasty of the profound femoral artery better blood flow and arrest of pains in rest were noted. Lumbar sympathectomy was performed in 142 (41%) patients, in 46 of them it was combined with continuous intraarterial infusion and in 20 patients -in combination with the rotor osteotrepanation on the femur and shin. Good results after sympathectomy were noted in 102 patients. PMID- 11011415 TI - [Infectious lesions of recurrent laryngeal and hypoglossal nerves as a rare complication of phlegmon of deep neck spaces]. PMID- 11011416 TI - [Late rupture of the left ventricular posterior wall with the formation of pseudoaneurysm after implantation of mitral and aortic valve prostheses]. PMID- 11011418 TI - [Treatment of a patient with a large defect of the vertical part of the duodenum and fistula formation]. PMID- 11011417 TI - [Perforating gunshot wound of the duodenum with injury of the inferior vena cava and multiple wounds of the jejunum]. PMID- 11011419 TI - [Hemicolectomy in a patient with hemophilia A]. PMID- 11011421 TI - [Certain general problems of surgery]. PMID- 11011420 TI - [Diagnostic tactics in localized fibroadenomatosis of the breast]. AB - Results of the examination and treatment of 203 patients with localized fibroadenomatosis of the mammary gland are described. The authors give an estimation of diagnostic algorithms used when detecting the physical signs of the disease. It was shown that the examination of patients with the palpation signs of nodular mastopathy should necessarily include mammography which can eliminate unwarranted risk of hypodiagnosing carcinoma of the mammary gland and determine the group of patients in whom the emergency operation is not necessary. PMID- 11011422 TI - [The 7th International Conference of Surgeons-Hepatologists of Russia and CIS countries (City of Smolensk, 14-16 November 1999)]. PMID- 11011423 TI - [Risk factors in the etiology of lung cancer]. PMID- 11011424 TI - [Injuries of extremity blood vessels]. PMID- 11011425 TI - [Incidence of high grade prostatic intraepithelial neoplasia in urologic practice]. AB - High grade (PIN AG) intraepithelial neoplasia of the prostate is a likely precursor of prostate adenocarcinoma (PA) because of their association. Since the risk to suffer PA increases in patients with no previous PIN AG, its finding requires an arduous search for PA. This paper reviews the incidence of PIN AG in 499 histological studies in prostate transrectal biopsies, prostate TUR and adenomectomy specimens and radical prostatectomy (RP) sections. Evaluation of data obtained, type of presentation and association to prostate carcinoma, indicating the approach taken in the various cases. PMID- 11011426 TI - [Prostatic carcinoma. Radical prostatectomy. Impact of neoadjuvant hormonal treatment]. AB - OBJECTIVE: To assess the influence of neo-adjuvant hormone therapy on the pathologic stage, margins and disease-free survival in patients with T1-T2 prostate cancer after radical prostatectomy. MATERIAL AND METHODS: Forty-two non randomised, T1-T2 patients who underwent complete neo-adjuvant blockade and radical prostatectomy between 1988-1998 were included in the study. This group is part of a larger cohort of 220 T1-T2 patients treated with radical surgery. The group is compared to: a) the remaining patients (178) (Group I) and b) a selected group (54 patients) of consistent characteristics relative to clinical variables (Group II). Mean follow-up: 35 months; median: 32 months. RESULTS: Compared to Group I patients with neo-adjuvant therapy show worse Gleason (p: 0.01), worse clinical stage (p: 0.02), higher rate of patients with high clinical risk (p: 0.03), and higher PSA (though not statistically different). No differences were seen relative to pathological status or margins. Compared to Group II (54 patients) no differences are found in clinical or pathological variables. Groups are comparable in terms of progression-free survival. CONCLUSIONS: Neo-adjuvant hormone therapy in high clinical risk, T1-T2 patients does not involve improvement in pathological stage, margins or progression-free survival. PMID- 11011427 TI - [Ureteroscopy as diagnostic and treatment method in ureteral processes: our experience]. AB - Endoscopic approach to the ureter both as a diagnostic procedure in various diseases and treatment of low ureteral lithiasis is a widely used technique. Review of case reports where this technique was used over the last four years. A total of 73 ureteroscopies were conducted, 64 as therapeutic approach (89.04%) and 9 for diagnosis (10.96%) achieving an overall 94.4% success rate. Multiple variables such as type and location of the stones, patient's age and sex, diagnostic methods, length of stay etc. All the above data were used for a descriptive analysis of the situation in our centre. In our experience, ureteroscopy is a good therapeutic option for low ureteral lithiasis, mainly when no extracorporeal lithotrity is available, and an excellent diagnostic tool in other ureteral processes when more traditional methods fail. PMID- 11011428 TI - [Morphologic course of prostatic hyperplasia]. AB - OBJECTIVE: To find a morphologic model of the development of prostate hyperplasia. METHODS: Study of the transitional zone in 60 patients (30 with infravesical obstruction, 30 with no obstructive symptoms) and quantification of the involved surface, number of pure stromal and glandular-stromal nodes, node area for each of them, and non-nodular area of the transitional zone, correlating each parameter to age based on clinical status. RESULTS: The greater transitional zone area is seen in patients with obstruction: 1376.83 +/- 408.17 mm2 vs 321.39 +/- 151.49 mm2 in asymptomatic patients, mainly due to a higher number of glandular-stromal nodes (17 vs 2.2) and their size, with a correlation to age (p = 0.03). Moderate increases of non-nodular areas are also found. CONCLUSIONS: These findings suggest that onset of prostate hyperplasia may be due to a consistent increase of the transitional area, and that in some patients, probably because of local factors, nodular development occurs as a result of both an increase in nodes number and size. PMID- 11011429 TI - [Radical prostatectomy in stage pT3C stage prostatic adenocarcinoma]. AB - OBJECTIVES: To assess the efficacy of surgery in pT3c patients; to discern whether this is an independent influential factor for survival and clinical factor for diagnostic prediction. MATERIAL AND METHODS: Forty-five pT3c (TNM 1982) patients from a cohort of 220 subjects with T1-T2 prostate cancer who underwent radical prostatectomy. Mean and median follow-up: 42 and 36 months, respectively. RESULTS: pT3 stage accounts for 20% (45/220) patients. They display a significantly higher mean PSA (25 +/- 2 ng/ml) (0.001), worse Gleason (0.0002) and clinical stage (0.0003), greater margins involvement (0.0007), and biochemical (0.02), local (0.05) and metastatic (0.001) progression. Independent influential factors are: PSA > 20 ng/ml; T2bc and Gleason 7-10. From these values patients can be divided into 3 risk groups: a) Group I (0-1 unfavourable variables): risk of seminal vesicle involvement 7%; Group II (2 unfavourable variables): risk 47%; Group III (3 unfavourable variables): risk 61%. Progression free survival: significantly lower than patients with pT2 (58 +/- 8% vs 66 +/- 6% at 5 years) (0.002), but similar to those with pT3ab (0.91). Seminal vesicle involvement is not an independent influential factor (Cox's multivariate study). Influential factors are PSA, Gleason and clinical stage. CONCLUSIONS: pT3c tumours are not an independently influential group in progression-free survival. Progression-free survival is lower than pT2 but similar to pT3ab. Their poor prognosis is dependent on association to negatively influential clinico pathological factors. PMID- 11011430 TI - [Survival in prostatic cancer, treated conservatively]. AB - OBJECTIVES: To analyse the survival rate among prostate cancer patients with conservative treatment. To compare survival between individuals with localised and invasive tumours, as well as between subjects below and above 70 years old at the time of diagnosis. METHODS: This study includes 80 patients diagnosed with prostate cancer between 1987 and 1988 at the Urology Service of the Central Hospital of Asturias (Spain). The cases were taken and followed retrospectively for 10 years. Kaplan Meter curves and Cox regression analyses were used for studying patients globally. Patients were categorised by tumour stage as well as by their age at the time of initial diagnosis. RESULTS: 10 patients remained living 5 years after diagnosis, corresponding to a 17% survival rate. However, 10 years after diagnosis, only 1 subject remained alive. Mean survival expressed in days was 677.28% of the cases died after the first year of diagnosis. Significant differences in survival rates were not found with regard to the age of patients ar the time of the diagnosis. However, survival is significantly higher among patients with localised tumours as opposed to invasive ones (p = 0.025; RR = 2.3). CONCLUSIONS: The survival rates is low in this study. The majority of tumours were at advanced stages when diagnosed and were mostly found in very old patients. Today, the use of PSA allows doctors to diagnose prostate cancer at earlier stages and in younger patients. This has lead to an increase in the frequency of radical prostatectomy. Future research should investigate whether this increase in radical prostatectomy has significantly changed the mortality rate of patients with prostate cancer. PMID- 11011431 TI - [Transrectal ultrasonography in the assessment of patients after radical prostatectomy. Normal and pathologic ultrasonography anatomy of vesico-urethral anastomosis]. AB - OBJECTIVE: To determine the ultrasound characteristics of the vesico-urethral anastomotic complex in patients with radical prostatectomy to separate normal prostate fossae from those affected by local recurrence or residual tumour. MATERIAL AND METHODS: Between January 1998 and June 1999 15 ultrasound guided transrectal biopsies of vesico-urethral anastomosis or prostate fossae were performed in patients with radical prostatectomy. The selection criteria to perform puncture in these patients were a negative extension study (abdominal pelvic CT and bone scan), PSA higher than 0.4 ng/mL and/or suspicious DRE. The study was completed with 10 transrectal ultrasound (TRU) after radical prostatectomy in patients with normal PSA and DRE. TRU parameters in patients with tumour-positive biopsies were compare to those from patients with negative biopsies and those obtained from TRU in patients with both normal PSA and normal DRE. RESULTS: The ultrasound parameters that best described prostate fossa abnormalities were the presence of hypoechoic masses or nodes and the integrity or not of the retro-anastomotic fat layer. When a hypoechoic mass or node was found at the anastomosis level, 80% biopsies were positive for tumour local recurrence. The percentage increases when changes are seen at the retro anastomotic fat layer. CONCLUSIONS: In spite of a good correlation between ultrasound abnormalities and positive biopsies, ultrasound findings from the vesico-urethral anastomosis in patients with radical prostatectomy are not well defined. Nodes or ultrasound irregularities can exist in normal prostate fossae as well as normal anastomosis ultrasound in the presence of tumoral relapses. We believe TRU of vesico-urethral anastomosis to be the best method available to evaluate local recurrence or tumoral persistence after radical prostatectomy. PMID- 11011433 TI - [Testicular embryonal carcinoma with contralateral synchronous intratubular germ cell neoplasia: analysis of a case]. AB - We report the case of a 20-year old male with a right testicular tumor. Bilateral orchidectomy was practised considering the synchronous clinical, ultrasonographical and histological (intraoperative biopsy) findings of the left testis. The definitive pathological report showed a right embryonal carcinoma with wide intratubular germ cell neoplasia (IGCN) of the contralateral testis. IGCN (formerly carcinoma in situ) is present in about 5% of cases in the contralateral gonad of those patients with a testicular neoplasm. More than 50% will develop cancer in that testis. Clinical and physical examination findings are usually unspecific. The diagnosis of IGCN is based on biopsy, although ultrasonography could give some data too, as some authors report. We analyze the therapy options for IGCN: (orchidectomy, chemotherapy, radiotherapy, or "wait and see"). In our case, the first one was made. Chemotherapy was used due to existence of retroperitoneal lymph node metastases, with an excellent follow-up afterwards. PMID- 11011432 TI - [Primary malignant melanoma of the female urethra. Report of a case]. AB - Primary malignant melanoma of the female urethra is a rare tumour with great agressivity and poor prognosis. An early diagnosis may benefit of radical surgery and adjuvant immunotherapy with curative effect. We present a sixty-five years old female with a malignant melanoma of urethra and a secondary additional melanoma in vulva, that was treated with both radical ureterectomy and vulvectomy. A bilateral inguinal lymphadenectomy and immunotherapy with alpha interferon were added. The patient is free of disease one year postoperatively. PMID- 11011435 TI - [Bladder compression by collateral veins]. AB - Many space-filling injuries (e.g. haematoma, abscess, aneurysm, lymphocele, neoplasia etc.) may result in extrinsic compression of the bladder. We contribute one case report of compression of the bladder from collateral venous circulation in a patient who came for a left varicocele. PMID- 11011436 TI - [Intraperitoneal bladder perforation in emphysematous cystitis]. AB - Contribution of one case report of a 69-year old diabetic male patient with neurogenic bladder. The patient developed emphysematous cystitis and peritonitis secondary to intraperitoneal perforation of the bladder. Emphysematous cystitis is an uncommon clinico-radiologic entity more frequent in diabetic and female patients and the elderly. Early diagnosis and treatment may avoid a fatal outcome. The paper includes a review of this condition. PMID- 11011434 TI - [Cystic ureteritis: importance of chronic infection-inflammation as etiologic factor. Report of a clinical case]. AB - Cystic Ureteritis is a very rare disease. It is more commun in elderly patients. This pathology is characterized by the formation of several submucous cystic in ureter, pelvis and/or bladder. We project the importance of the infectious and inflammatory factors as the disease etiology. We report a case of a woman with recurrent urinary tract infection, coral lithiasis and left ureteritis cystica. It is included a literature review and we explain the main clinical, diagnostic and therapeutic aspects. PMID- 11011437 TI - [Wolfram syndrome. Urologic implications]. AB - OBJECTIVES: We report on four new cases of Wolfram's Syndrome. We emphasize in urological aspects of this disease. PATIENTS AND METHODS: Three male siblings. The other patient is also a male, without familiar relation with the other ones. All four patients presents different levels of urological alterations, mainly urinary collecting system dilation and decrease in detrusor muscle contractility. CONCLUSIONS: Urological findings are cardinal aspects in Wolfram syndrome. Due to its high frequency and prognostic value in natural history of disease. Urological disease seems to be within a systemic neurological tissues affectation of etiology that remains unknown. PMID- 11011439 TI - [Gonadal thermoregulation in aquatic mammals. An evolutionary guarantee for the preservation of the species]. PMID- 11011438 TI - [Giant inguinoscrotal bladder hernia]. AB - Inguinoscrotal bladder hernia occur in 0.4 to 3% of general poblation, though massive hernia are much rare. We present a new case of a giant inguinoscrotal bladder hernia, which was solved by surgery. It includes a clinical, diagnostic and therapeutic aspects, and a review of the relative literature. PMID- 11011440 TI - [Effect of sacral roots block in the prostatic structure of the rat]. AB - OBJECTIVE: To validate a simplified model of neural manipulation of the lower urinary tract of the rat, by means of alcoholic blockade of sacral roots. MATERIAL AND METHODS: Twenty Sprague-Dawle rats (aged 3 months) underwent alcohol sacral root blockade under total anesthesia with previous needle placement monitoring (electrostimulation). After 28-30 days, ipsilateral ventral prostatic lobe (VL) was obtained. Macroscopic and light microscopy (LM) studies were performed (computerized image analysis). Results were compared with 20 LV from non manipulated rats. RESULTS: A slight non significant decrease of LV weight in the blockade group was notice. LM study showed a reduced epithelial height after manipulation. Gland compartment mean proportions were: control group: 28.1% (epithelial), 29.8% (stromal), y 70.2% (glandular -lumen plus epithelium-); study group: 30.5% (not significant), 37.4% y 62.3% (both p < 0.05) respectively. Epithelium area did not suffer any variation. A reduced vascular overall count was noted in the study group. Ipsilateral diffusion of blockade solution was shown in 9 rats (45%), and bilateral in 11 (55%). CONCLUSIONS: Alcoholic sacral root blockade produces an atrophy of the glandular component (due to luminal shrinkage, without epithelial change). No differences were found after the macroscopic study. This study could not reproduce the changes produced after standard surgical denervation, therefore we cannot recommend this method to be used in future models. PMID- 11011442 TI - [Relationship between tumor recurrence and the expression of the p53 gene in primary superficial transitional carcinoma of the bladder]. AB - Contribution of 60 patients with primary surface transitional cell tumours of the bladder where nuclear expression of p53 protein was prospectively studied and compared to known prognostic factors in an attempt to find out its role in the development of relapses. An statistically significant relationship was found between the protein expression and cytology, tumoral multifocality, stage, relapse development and tumoral progression. It can be concluded that expression of this protein can be of use as relapse predictor. PMID- 11011441 TI - [Initial superficial bladder tumors: our experience. Our criteria. Study Group of Bladder Tumors]. AB - OBJECTIVES: Evaluation of a homogeneous series of Superficial Primary TCC of the Bladder. Utility of randomized biopsies (BMN). Study of risk factors and risk groups as a basis for determining treatment and followup patterns. MATERIALS AND METHODS: Survey of 1,529 PSBT cases that underwent TURB and BMN, with a mean followup of 4.2 years. Strict statistical treatment: multivariate analysis using Cox logistic regression according to Kaplan-Meier curves. RESULTS: Tumor features of note: single tumor (65%), smaller than 1.5 cm (45%), high grade (G3: 26%), and high stage (T1: 64%). BMN showed associated CIS in 284 cases (19%). Main risk factors for recurrence were multiplicity, size > 3 cm and association with CIS. The main risk factor for progression was grade 3 (OR 19.9). Grade 3 and association with CIS were found to increase mortality risk. On the basis of this data, we grouped tumors according to low-, middle-, and high-risk, and established our proposed treatment and followup for each group. CONCLUSIONS: This multivariate analysis of a homogeneous cohort of 1,529 primary superficial bladder tumor cases allowed vs to determine risk groups for treatment and followup. PMID- 11011443 TI - [Neoadjuvant chemotherapy MVAC in the treatment of infiltrating bladder carcinoma]. AB - INTRODUCTION: Aiming to preserve the bladder in patients with infiltrative carcinoma of the bladder and to offer patients improved quality of life with no detriment for survival, a therapeutical protocol was set up. MATERIAL AND METHODS: Between August 1988 and January 1997 63 patients with stage T2 and T3a infiltrative carcinoma of the bladder, with no metastasis or node extension detectable with imaging techniques were treated in our unit. 45 of these patients met all protocol criteria and were given 3 neoadjuvant chemotherapy courses with MVAC (methotrexate, vincristine, adriamycin, cisplatin). INCLUSION CRITERIA: age under 75 years, Karnofsky greater than 50%, leucocytes greater than 2,500 cell/mL and platelet greater than 100,000/mL. Following chemotherapy, re-assessment was performed through lab tests, chest X-rays, abdomino-pelvic CT, bone scanning, cystoscopy, multiple randomized biopsies, tumoral bed scar resection and resection of relapsed urothelioma. Patients with complete remission were given radiotherapy. Those showing stabilisation of progression were proposed to undergo cystectomy. Fisher's test or chi 2 test were used for the comparison of qualitative variables. The survival analysis was performed using the Kaplan-Meier method. The curves comparison was done with Breslow's exact test. A Cox's proportional risk method allowed to calculate the relative risks together with their 95% confidence interval. RESULTS: 53.7% patients included in this protocol showed complete remission, 41.5% stable disease and 4.9% progression. 62.2% of patients were given radiotherapy versus 17.8% who underwent cystectomy. 20% received other therapies after rejecting both cystectomy or radiotherapy. Median follow-up was 43.38 months. Overall median survival was 96 months. The accumulate probability of survival at 4 years was 79%. 50% patients with complete clinical response relapsed during follow-up. Tumoral stage of those who relapsed was lower than the initial one in 63.7% cases, remained the same in 18.2%, and higher in 18.2%. With regards to grading, 66.7% patients had lower grading at relapse if tumour was initially grade 2. For those with initial tumour grade 3, only 20% had a lower grade. CONCLUSION: 64.4% patients retained their bladder. In 26.7% there was demonstrable metastatic disease. No differences were seen in the distribution or survival time based on the different treatment given after chemotherapy (p = 0.22). Patients with complete remission after chemotherapy have greater actuarial survival which is statistically significant (p = 0.04). PMID- 11011444 TI - [N1 prostatic adenocarcinoma treated with radical surgery and immediate hormonal management]. AB - OBJECTIVES: To study the survival in patients with prostate adenocarcinoma, lymph nodes involvement in the intraoperative pathological examination, evaluating both radical surgery and early ablation as treatment. Identification of clinical factors that can predict node involvement. METHOD: 20 patients who underwent radical prostatectomy between 1988-1998 were included in the study. All patients clinically diagnosed with T1-2N0Mx prostate adenocarcinoma (T.N.M. 1992), single metastatic node involvement in the pathological study and early sub-albuginea orchidectomy. This group was compared to a 200-patient control group also with T1 2N0Mx prostate adenocarcinoma but with no pathologic nodular involvement. Statistical study: contingency 2Xb tables (Pearson's chi 2 or Fisher's exact test) to compare qualitative variables; Student's t test for means comparison; Kaplan-Meier for actuarial survival calculations and comparison of Log-rank survival curves. For the study of clinical variants with influence in node involvement a logistic regression model was used. RESULTS: Mean age was 63 +/- 0.8 years. Median follow up 56 months. Mean PSA 33 +/- 4.4 ng/mL and 55% had Gleason 5-7. 16 patients were stage pT3. Specific 5-year survival was 90 +/- 8% and biochemical progression free survival 63 +/- 12%. Clinical variables with statistical significance for node involvement are: pre-surgical PSA greater than 20 ng/mL (RR = 4.6), and Gleason higher than 4 (RR = 3). The remaining variables showed no statistical significance. CONCLUSIONS: Gleason and PSA are the only clinical values that predict node involvement. The procedure performed obtains good results and a survival comparable to that of the control group. PMID- 11011447 TI - [Percutaneous nephrostomy as management technique of obstructive uropathy: review of 105 cases]. AB - 105 percutaneous nephrostomies in 73 patients. Indication for nephrostomy was obstruction in the drainage system in all cases. Nephrostomy was bilateral in 24 cases. Reasons for nephrostomy were: lithiasis in 35 cases, carcinoma of the bladder in 17, post-surgical iatrogenic stenosis in 13, prostate carcinoma in 9, cancer of the rectum in 9, infection in 5, neoplasia of the upper urinary tract in 5, retroperitoneal fibrosis in 3, glandular cystitis cystica in 3, ovarian cancer in 3, congenital in 2, lymphoma in 1. Six case were single-kidney patients. Renal puncture through the lower calyceal group was the preferred approach. Purulent urine was extracted in 10 cases. Complications of nephrostomy included haematuria in 34 cases, contrast extravasation in 8, fever in 6, 1 case of death due to septic shock, 2 perirenal haematoma (nephrectomy was required in 1 case), and catheter detachment in 5 cases. All patients showed improvement of both clinical signs and symptoms, and lab results. PMID- 11011445 TI - [Diagnostic algorithm based on a multivariate model to reduce the percentage of negative prostatic biopsies in patients with changed PSA or suspicious rectal palpation]. AB - OBJECTIVES: To determine what tests have a better diagnosis utility in patient with suspected prostate cancer due to PSA equal or greater than 4 ng/ml or abnormal digital rectal examination in order to reduce the number of negative prostate biopsies. MATERIAL AND METHODS: We carried out a ultrasound-guided sextant transrectal biopsy in a series of 400 patient with prostate-specific antigen (PSA) levels equal or greater than 4 ng/ml or abnormal digital rectal examination. All patients had also transrectal rectal ultrasonography to value the echographic prostatic characteristics and the prostatic volume, and a free PSA determination. RESULTS: The free/total PSA ratio (PSAl) and the PSA density (DPSA) were the most powerful predictors of prostate cancer, both with a 66% sensitivity and a 70% specificity (at a 0.15 cutoff), followed by the total PSA (PSAt), the digital rectal examination and the hypoechogenic prostatic nodules. We constructed a logistic multivariate model with these data. The outcome variable of logistic model was the probability of having prostate cancer. The significant predictive variables of the model were the PSAl, the DPSA, the digital rectal examination and the presence and extension of hypoechogenic prostatic. This model had a 81% sensitivity and 79% specificity at a 0.24 probability cutoff. We considered a 0.1 probability cutoff to reduce the number of false negative. With this strategy the sensitivity was 94% and the specificity 54%. If we had applied this model to the patient of our series then, we would be able to avoid prostate biopsy in 164 cases (the 41% of the patient), leaving only 7 cases without diagnosis of prostate cancer. CONCLUSIONS: The most effective combination parameters were the digital rectal examination, PSAl, presence and extension of prostatic hypoechogenic zones and DPSA. This combination allows us to diminish the percentage of negative prostate biopsy in patient with prostate specific antigen (PSA) levels equal or greater than 4 ng/ml or abnormal digital rectal examination without significantly descending the number of detected prostate cancers. PMID- 11011446 TI - [Prostatic rebiopsy. Prognosis factors of the anatomopathologic result]. AB - The indications for repeat prostate needle biopsy after a previous biopsy are not defined. We examined 107 prostate biopsies (in 98 patients) without a diagnosis of malignancy, which we repeat. Carcinoma was detected in 31 patients (31.6%). We didn't find statistic relationship between the repeat biopsy's outcome and: interval between biopsy and repeat biopsy, PSA value, PSA density (biopsy), PSAD of the transitional area (PSAD ad., on repeat biopsy). We found relationship with: prostatic weight (p = 0.002 on the biopsy, p = 0.0002 on the repeat biopsy), volume of the transitional area (p = 0.02 on the biopsy, p = 0.0001 on the repeat biopsy), PSA value (p = 0.02, on the repeat biopsy), PSAD ad. (p = 0.002, on the repeat biopsy), and with PSA velocity (p = 0.008). We only found clinic usefulness for the PSA velocity: patients with PSA velocity greater than 1 ng/ml/year are at high risk for prostate carcinoma on the repeat biopsy, specially in small prostates. PMID- 11011448 TI - [Primary bladder actinomycosis]. AB - Primary actinomycosis of the bladder is an infrequent presentation of this condition. The main concern is obtaining a differential diagnosis from tumours of the bladder and the subsequent impact this has on the therapeutic choices. Contribution of one case report of a male patient with actinomycosis of the bladder, with remarks on relevant diagnostic (including differential diagnosis) and therapeutic data. PMID- 11011450 TI - [Malignant fibrohistiocytoma of the bladder]. AB - We report on new case of a rare vesical tumour. We result the importance of immunohistochemistry and ultrastructural study to support the diagnosis of malignant fibrous histiocytoma of the urinary bladder. There has been described another 16 cases of this tumour in the literature, however, only four of them five with ours- reports an extensive immunohistochemical and ultrastructural study. The rarity of vesical localization of this tumour may delay its diagnosis. It must be supported by a immunohistochemistry and/or ultrastructural study, in order to differentiate from other tumours with fibrohistiocytoma-like pattern: leiomyosarcoma and sarcomatoid carcinoma of the bladder. After radical removal of tumour, adjuvant therapy is recommended both systemic chemotherapy and local radiotherapy, although survival rates are over 5.3 months after first therapeutical actuation. PMID- 11011449 TI - [Renal malacoplakia. Report of a new case]. AB - The malacoplakia is an infrequent granulomatous inflammatory disease which pathogenesis is not well understood and affects predominantly the genitourinary system, through every organ can be affected. We present a case of unilateral renal malacoplakia in a male that made ist debut with septic shock and perinephritic abscess. It was operated under the suspicious diagnostic of renal neoplasm. The histologic study grave us the diagnosis of renal malacoplakia extending until the skin. After nephrectomy and medical treatment the patient had an uneventful course. PMID- 11011451 TI - [Technical solution in kidney retransplantation in a patient with femoro-femoral bypass]. AB - Over the last decade kidney transplantation has become a common therapeutic procedure for patients with end-stage renal diseases. Between 1988 and 1998 donors rate per million population has dramatically increased in our environment, thus providing us with more chances to offer kidney transplantation to a larger number of patients. The technico-surgical difficulties that have to be faced however, are increasingly frequent and require a search for new approaches and innovative changes to the usual surgical techniques for our patient's benefit. PMID- 11011452 TI - [Treatment of pyelocaliceal transitional cell carcinoma with partial nephrectomy]. AB - Nephroureterectomy is the choice treatment for tumours of the upper urinary tract. Conservative surgical management is sometimes warranted due to the risk of renal failure or just as palliative treatment. Contribution of two cases of transitional tumours in the pyelocalyceal zone treated with partial nephrectomy, and discussion of the usefulness and indication of conservative surgery for these tumours. PMID- 11011453 TI - [Adrenal cortex carcinoma with right atrium involvement. Surgery with cardiopulmonary bypass]. AB - INTRODUCTION: Primary adrenal carcinoma is a low incidence tumour. About 50% are functional and the majority result in Cushing's syndrome, while detection of the rest is commonly incidental. Surgical management is through radical surgery. CASE REPORT: A 36-year old female who presented with a one-month old abdominal pain in the hypochondrium and the right lumbar fossa. Physical examination found a discretely painful mobile mass. Adrenal hormones values were normal. Ultrasound and CT studies detected a 14-cm adrenal tumour with cava vein thrombosis up to the right auricle. The surgical procedure included laparotomy, liver mobilisation and cardiopulmonary by-pass. The thrombus was completely removed by auriculotomy. Therapy with mitotane and 5-fluorouracil was then instituted. Thirteen months after surgery the patient is asymptomatic and tumour free. REMARKS: Dissemination of adrenal tumours to veins ranges between 6 to 30%. Clinical manifestation of cava vein and auricle involvement is variable but it may develop with no symptoms due to collateral circulation. Pre-operative diagnosis is critical to plan for adequate surgical approach. Ultrasound and computerised tomography can adequately diagnose the process, but nuclear magnetic resonance provides more information on the thrombus extension and location. In the absence of node spread, local extension, or invasion of the cava wall prognosis is similar to that of patients with no vein involvement. The best therapeutic option is surgery with removal of the lesion and cardiopulmonary by-pass. Survival at 5 years is 43%. Post operative administration of chemotherapy with mitotane is a useful and recommendable choice. PMID- 11011454 TI - [Glandular cystic cystitis]. AB - Cystic-glandular cystitis is considered as part of the urothelial pre-neoplastic proliferative abnormalities. This group includes atypical hyperplasia. Von Brunn's nidus, and cystitis cystica. They are a consequence of the changes experienced at the urothelium level in response to inflammation, irritation or carcinogens. Diagnosis is mainly based in the pathoanatomical study of the biopsy obtained following endoscopic resection. The signs and symptoms it presents are varied and show a clear relationship to distribution and extension of cysts. Treatment is based in the removal of irritative factors. Cystectomy with urinary by-pass may be necessary if required by clinical evolution. PMID- 11011456 TI - Immunisation in the age of the human genome. PMID- 11011455 TI - [Relationship between preoperative urine cultures and prostatic gland cultures in patients surgically treated for prostatic benign hyperplasia]. PMID- 11011457 TI - The investigation of a 'cluster' of hepatitis B in teenagers from an indigenous community in North Queensland. AB - BACKGROUND: In early 1999, five teenagers from the same Indigenous community were notified as having hepatitis B. Hepatitis B vaccine should have been offered to this cohort of teenagers in a 'catch-up' program during the late 1980s when they were of pre-school age. OBJECTIVES: To determine the vaccination status of residents of the community born between 1981 and 1985 (inclusive) and to ascertain the prevalence of markers of hepatitis B infection and carriage in the incompletely vaccinated teenagers in this cohort. METHODS: Community health records were examined to identify all residents in the study cohort. Immunisation records were obtained from local hospital records and from a statewide computerised vaccination database. Serological tests for markers of hepatitis B infection and carriage were performed on blood samples from the incompletely vaccinated teenagers. RESULTS: Only 44% of 235 teenagers who had their vaccination status assessed were fully vaccinated. One hundred and eleven (47%) of the cohort had not received any hepatitis B vaccine. Over 90% of the incompletely vaccinated had been infected with the hepatitis B virus and 26% of these were hepatitis B carriers. CONCLUSIONS: Despite the availability of an effective hepatitis B vaccine and the recommendation for a catch-up program, the pre-school aged cohort of children at the community were not effectively targeted for vaccination. Hepatitis B remains a consequential infection in Indigenous communities in North Queensland. IMPLICATIONS: Initiatives to control hepatitis B need to be enhanced within existing maternal and child health, sexual health, alcohol and drug and chronic disease management programs. PMID- 11011458 TI - Health promotion and older people: the role of the general practitioner in Australia in promoting healthy ageing. AB - OBJECTIVE: The Commonwealth Government's Enhanced Primary Care initiative supports measures to enhance the role of general practitioners (GPs) in promoting healthy ageing as part of a population health approach. This paper comments on how the health assessments can be conducted to best effect, to strengthen the role of GPs in primary care and to promote autonomy and independence in older people. METHOD: The relevant literature was collated to produce a review of public health and health promotion approaches and to ascertain the effectiveness of health promotion interventions for older people. A broad definition of health promotion including primary, secondary and tertiary prevention was adopted. RESULTS: The evidence base suggests there is scope for greater targeting of health promotion activities towards older people. The rationale for the Australian GP to assume a major health promotion role with their older patients is provided. Associated barriers and enablers are discussed. CONCLUSIONS: Prevention of disability is a key public health issue. The new MBS items may enable systematic evaluation of function and assist healthy ageing for all older people, including the frail aged. An increase in the preventive advice given to older patients has the potential to increase healthy behaviours and alter health outcomes. IMPLICATIONS: The annual health assessment items on the Medicare Benefits Schedule, by enabling the GP to focus on prevention and coordination of care, have the potential to improve the health, physical, psychological and social function of older Australians. PMID- 11011459 TI - Response rates of Victorian general practitioners to a mailed survey on miscarriage: randomised trial of a prize and two forms of introduction to the research. AB - AIMS: To identify whether the form of introduction to a study and knowledge of a substantial prize influence the response rate of general practitioners (GPs) to a postal survey. METHODS: A postal survey of 700 randomly selected Victorian GPs concerning management of early pregnancy bleeding and miscarriage, incorporating two randomised-controlled trials of recruitment methods; analysis of response rates and costs at 4 weeks and 11 weeks. RESULTS: The response rate was 61.5% of eligible participants. Doctors made aware of a prize were more likely to respond in the first four weeks (difference in response rate 10.2%, 95% confidence interval (CI) 2.8%-17.6%). This difference diminished after the first four weeks. Doctors introduced to the survey by a telephone call were no more likely to respond than those introduced by a postcard. The use of a postcard saved 73% of the cost of introducing the survey by telephone. Female doctors were more likely than males to reply (difference 12.3%, 95% CI 4.7%-19.9%). Rural doctors were no more likely to reply than urban doctors. Very few doctors (16.2%) completed a Practice Assessment activity associated with the survey. CONCLUSIONS: A valuable prize will accelerate response to a survey by GPs, thereby reducing the costs of follow-up. The cost of telephoned introductions is not justified, when compared with a brief written introduction. PMID- 11011460 TI - Socioeconomic correlates of mortality differentials by local government area in rural northern New South Wales, 1981-1995. AB - OBJECTIVE: Many studies have examined the correlation between socioeconomic status (SES) and mortality in Australia, but little is known about the correlation in rural areas and most studies have not explored the trends in SES differentials in mortality. This ecological study examines this correlation and explores the impact of the national strategies to reduce SES differentials in mortality in a rural area. METHODS: Mortality data for residents in the New England Health Area, New South Wales (NSW), 1981 to 1995, were analysed. Twenty Local Government Areas (LGAs) in New England were ranked and aggregated into 4 groups according to a composite SES indicator from the 1996 census, and age/sex adjusted mortality rates were calculated for each group and compared. Poisson regression models were used to assess the linear trends in mortality for 1981-95. RESULTS: A strong relationship between working age adult mortality and SES was found for both sexes. The rates for the most disadvantaged LGAs were significantly higher than the least disadvantaged LGAs for both sexes. The mortality rate was consistently higher for the most disadvantaged LGAs than the least disadvantaged LGAs. CONCLUSION AND IMPLICATION: Although there has been an overall decline in death for all 4 groups of LGAs, the gap between the most disadvantaged and the least disadvantaged groups has widened over the last 15 years. This widening gap in death rates suggests that the strategies implemented as part of the Health for All initiative to reduce inequalities in mortality differentials have not been effective in this rural area. PMID- 11011461 TI - More or less equal? Comparing Australian income-related inequality in self reported health with other industrialised countries. AB - OBJECTIVE: To measure the distribution of self-reported health by income in order to compare the level of health inequality in Australia with other industrialized countries. METHOD: Using data from the two National Health Surveys undertaken in 1989-90 and 1995, concentration indexes were calculated to quantify the distribution of self-reported health by equivalent income. The concentration index for Australia was compared with those reported for nine industrialized countries in Europe and North America. RESULTS: The estimated income-related concentration indexes were -0.1172 in 1989-90 and -0.1094 in 1995. CONCLUSION: The level of health inequality is not significantly different from the US or the UK, but significantly greater than seven other European nations. IMPLICATIONS: Australia has significant income related health inequalities and the distribution of health appears to be more unequal than in many other industrialized nations. There is a need to further investigate and quantify those features of the anglophone societies that set them apart from some other industrialized nations. PMID- 11011462 TI - Queenslanders' use of personal strategies to minimise risk of mosquito-borne disease. AB - OBJECTIVE: To describe Queenslanders' awareness, knowledge and attitudes towards mosquito-borne diseases and their transmission, and to determine which factors influence the adoption of effective individual prevention strategies. METHODS: In 1995-6, cross-sectional surveys of adult residents in the western suburbs of Brisbane and registered voters in Cairns were conducted. Forced entry logistic regression was used to predict use of personal protection and elimination of domestic breeding sites in the two cities. RESULTS: Final sample sizes were 347 in Cairns and 165 in Brisbane with response rates of approximately 70%. RRVD awareness was nearly universal in both cities. A majority of residents (60% in Brisbane and 65% in Cairns) report they are careful to avoid being bitten by mosquitoes. 25% of Cairns residents and 18% of Brisbane residents report always using some method of personal protection. Cairns residents are also more likely to say that they actively prevent mosquitoes from breeding in their yards (76% in Brisbane and 87% in Cairns). Knowledge of mosquitoes and disease transmission was slightly higher in Cairns. In Brisbane, dislike of mosquitoes and being regularly bitten were significant in the multivariate model predicting personal protection, whereas concern for disease and being female were significant in Cairns. Concern about disease was a significant predictor of eliminating breeding sites in both cities. CONCLUSIONS: Raising concern about mosquito-borne disease can increase use of personal prevention strategies. However, providing information on prevention strategies may not be effective. The most effective strategies are not practiced or seen by the public to be related to minimising risk of disease. IMPLICATIONS: Greater emphasis in health promotion campaigns should be placed on encouraging permanent alterations to the domestic environment rather than temporary methods that are difficult to sustain and not effective against the common vectors for mosquito-borne diseases in Queensland. Educational messages should explicitly link preventive behaviours with the reduction in the likelihood of contracting a serious disease. PMID- 11011463 TI - Collecting and using aboriginal health information in New South Wales. AB - OBJECTIVE: To describe the development of guidelines for the management of Aboriginal health information in NSW. The purpose of the guidelines is to promote the ethical management of Aboriginal health information, with appropriate consideration for cultural factors. METHODS: The guidelines were developed collaboratively by the NSW Aboriginal Health Partnership, which comprises NSW Health (the central administrative office, named NSW Department of Health, and the Area Health Services) and the NSW Aboriginal Health and Medical Research Council (AHMRC), the peak body representing member Aboriginal Community Controlled Health Services in NSW. A lengthy and comprehensive consultation process enabled a wide range of interested groups to have input into the guidelines. RESULTS: The project culminated in the production of the NSW Aboriginal Health Information Guidelines, covering the collection, ownership, storage, security, release, usage, interpretation and reporting of information, as well as issues of privacy and confidentiality. The Guidelines formed the basis of a formal Memorandum of Understanding, signed by the NSW Minister for Health, NSW Department of Health and the AHMRC, on 24 August 1998. CONCLUSIONS AND IMPLICATIONS: The Guidelines make an important contribution to meeting a need for protocols on the collection, ownership and use of Aboriginal health information. Their production reflects successful collaboration between government and Aboriginal Community Controlled Health Services in NSW. Future reviews of the Guidelines will ensure their effectiveness and consistency with Aboriginal community principles. PMID- 11011464 TI - Reproductive health, infertility and sexually transmitted infections in indigenous women in a remote community in the Northern Territory. AB - OBJECTIVE: To investigate markers of reproductive health in a remote Indigenous community in Northern Australia. METHODS: A retrospective, cross-sectional analysis of case notes of 342 women between the ages of 20 and 45 years, living in one community in a remote region of the Northern Territory. RESULTS: The total rate of current infertility in the community was 26.3%; 8.2% for primary infertility and 18.1% for secondary infertility. An additional 3.3% of women had resolved infertility. Only 43% of the women had sought medical help for the problem. A history of ectopic pregnancy was recorded in 2.6%, stillbirth in 1.8%, miscarriage in 14.3% and neonatal death in 12.3%. Depot steroidal contraception or tubal ligation were used by 50% of the women but 45.9% used no contraception. A history of pelvic inflammatory disease (PID), T. vaginalis N. gonorrhoeae, genital C. trachomatis infection, syphilis or bacterial vaginosis was noted in 32%, 46%, 27%, 30%, 41% and 9% respectively. Current alcohol consumption was reported in 23% and cigarette smoking in 76%. In multivariate analysis, infertility was strongly associated with PID (adjusted OR 8.5), alcohol consumption (AOR 3.1), T. vaginalis (AOR 2.5), N. gonorrhoeae (AOR 2.2) and bacterial vaginosis (AOR 2.9). CONCLUSION: Reproductive health is poor in this community of Indigenous women, with endemic levels of STDs, PID and tobacco consumption. The absence of barrier contraception (e.g. condoms, diaphragms) has implications for HIV and STD control. Clinical and public health interventions are urgently required but the implementation of these is hindered by a number of structural, social and economic barriers. PMID- 11011465 TI - Drowning and alcohol in New Zealand: what do the coroner's files tell us? AB - OBJECTIVE: To provide a systematic review of the details on alcohol involvement available in the coronial files to determine if there is enough evidence to estimate the role of alcohol in drowning. METHOD: We reviewed the coroner's files of persons 10 years or older who drowned in New Zealand between 1992-1994 inclusive. RESULTS: A total of 320 coroner's files were examined. Blood Alcohol Concentrations (BACs) were taken in 115 cases (36%) and positive for 50% of these. When accounting for the incomplete testing by using all the information on alcohol involvement collected, between 30-40% of the cases were estimated to have a positive BAC and between 17-24% to have a BAC 100 mg/dL or higher. CONCLUSION: The quality and completeness of current coronial information on alcohol involvement in drowning is insufficient to arrive at an accurate estimate of the percentage of drownings where alcohol was a factor. IMPLICATIONS: Coroners should test drowning victims 10 years and older for BAC. These data should be systematically recorded and processed with the goal of determining who should be targeted in drowning and alcohol prevention programs. PMID- 11011466 TI - The pattern of diabetes care in New South Wales: a five-year analysis using Medicare occasions of service data. AB - OBJECTIVE: To use Medicare occasions of service data to establish the pattern and standard of care received by people with diabetes. METHOD: Information about visits to medical practitioners as well as utilisation of diabetes related procedures for people living in New South Wales (NSW) for the individual years between 1993 to 1997 was retrieved using a Health Insurance Commission data file. Individuals were deemed to have diabetes if an HbA1c which can only be ordered for a person with known diabetes, had been performed over the five-year period. RESULTS: On average over the study period, persons with diabetes accounted for 3.1% of the population but they used 5.5% of general practitioner services. A large proportion of patients also received care at the specialist and consultant physician level, 51.2% and 38.6% respectively, a three to four fold increase when compared with their non-diabetic counterparts. There was also a 1.3 to 1.8 fold increase in the mean number of attendances to the various medical practitioners. Surveillance of diabetes parameters was inadequate but small improvements were seen over the 5 year study period (proportion of persons with diabetes with a HbA1c performed: 48.8% to 56.8%; Lipids: 49.4% to 52.0%; HDL cholesterol: 18.3% to 18.8%; microalbuminuria: 4.7% to 11.6%). CONCLUSION: This study has highlighted the heavy burden imposed by diabetes on our health care system. IMPLICATIONS: The use of Medicare occasions of service data represents a cost efficient way of monitoring health service utilisation. PMID- 11011467 TI - Sites for depression on the web: a comparison of consumer, professional and commercial sites. AB - OBJECTIVE: To provide information on the range of treatments for depression provided by commercial, professional and consumer web sites. METHODS: An audit of the 21 most popular depression sites on the world wide web undertaken from March 1999 to July 1999, in Canberra, ACT. Treatment types and categories of treatment were compared among commercial, professional and consumer web sites. RESULTS: A total of 53 treatments or treatment types were mentioned. The number of treatments mentioned per site ranged from 2 to 38. Antidepressant medication and psychotherapy were noted by almost all sites. Consumer sites mentioned psychological therapies less frequently but did not mention dietary supplements or complementary treatments more frequently. CONCLUSIONS AND IMPLICATIONS: Consumer web sites provide a point of comparison to those from commercial or professional sources. They provide a voice to inform psychiatrists and GPs about preferred treatments and side effects and to alert researchers to areas that need formal investigation. PMID- 11011469 TI - A workforce survey of health promotion education and training needs in the state of Victoria. AB - OBJECTIVE: This study investigates the education and training needs of health professionals and factors affecting participation in education and training. METHODS: A survey of health promotion professionals, health professionals, GPs and CEOs of community health centres, conducted across different settings and locations. Information was obtained on: involvement in health promotion activities, most useful content and format of past training, current preferences for education and training and barriers and incentives to education and training. RESULTS: Health promotion professionals were involved in the widest variety of health promotion activities, including more evaluation, research and planning than GPs and other health professionals who were involved in more client-focussed activities. Professionals' preference for training content reflected the type of activities in which they were most frequently involved. Practical courses, of short duration, delivered by experienced peers or health promotion experts were preferred over university and TAFE courses. Professionals in rural and provincial locations require both greater access to information on training and conveniently located training. More organisational support, funding and time release would encourage the training of professionals in government departments, community health centres and public hospitals. CONCLUSIONS: To be most effective, training must be tailored to suit the specific needs of different professionals involved in health promotion and take into consideration how factors, such as financial incentives and time release, influence participation across different settings and locations. IMPLICATIONS: Further development of the health promotion workforce will require recognition of its professional diversity and a more responsive and organised approach to education and training programs. PMID- 11011468 TI - Sharing obstetric care: barriers to integrated systems of care. AB - OBJECTIVES: To map the provision of shared obstetric care in Victoria, and investigate the views of care providers about the ways in which current practice could be improved. METHOD: All Victorian public hospitals with > or = 300 births per annum and a purposive sample of hospitals with < 300 births per annum were mailed a questionnaire seeking information about current practice. Interviews with key informants (n = 32) were conducted at four case study sites. RESULTS: The response rate to the hospital survey was 98% (42/43). Fourteen different models of shared care were identified. Two-thirds of hospitals with > or = 300 births per annum (16/28) had three or more different models of shared care. Six hospitals (15%) had written guidelines for all models of shared care offered; 13 (32%) had written guidelines covering some models. Practice varied considerably in relation to: exclusion criteria, recommended schedule of visits and use of patient-held records. There was little consensus about the content of visits and responsibility for covering particular aspects of care. Few hospitals (6/42) had written information for women about shared care. Care providers expressed divergent views regarding the question of where ultimate responsibility lies for individual patient care and for the overall management of shared care. CONCLUSIONS: Current funding arrangements provide strong incentives to expand enrollment in shared obstetric care. Expansion of shared care has occurred without the development of formal, consultative and agreed arrangements between providers, or adequate provision for monitoring, evaluation and review. The variety, complexity and fluidity of models of shared care and lack of agreed procedures contribute to difficulties experienced by both providers and women participating in shared care. IMPLICATIONS: Detailed evidence-based agreed guidelines developed in consultation with hospital and community providers, and provision of improved information to women about what to expect in shared care arrangements are urgently required. PMID- 11011470 TI - Cigarette smoking, menstrual symptoms and miscarriage among young women. AB - OBJECTIVE: To examine associations between cigarette smoking and menstrual symptoms and miscarriage among young women. METHOD: The study sample consists of 14,779 women aged 18-23 years who participated in the mailed baseline survey for the Australian Longitudinal Study on Women's Health, conducted in 1996. The main outcome measures are self reported menstrual symptoms and miscarriages. RESULTS: Current smokers and ex-smokers had an increased risk of menstrual symptoms and miscarriages compared with women who had never smoked, with the highest risk occurring in heavy smokers (adjusted odds ratios for those smoking > or = 20 cigarettes per day: premenstrual tension 1.5 (95% confidence interval 1.3 to 1.7), irregular periods 1.5 (1.3 to 1.8), heavy periods 1.6 (1.4 to 1.9), severe period pain 1.5 (1.4 to 1.7), one or more miscarriages 2.0 (1.5 to 2.8). The odds ratios generally increased with numbers of cigarettes smoked and a younger age of starting to smoke. CONCLUSION: This study suggests that young women who smoke are at higher risk of a range of menstrual problems and miscarriage than those who have never smoked. The immediacy of this risk (in contrast to the longer term risks of chronic disease) can be used to improve the relevance of anti-smoking campaigns targeted to young women. PMID- 11011471 TI - Cohort trends in the age of initiation of drug use in Australia. AB - AIM: To examine birth cohort trends in the prevalence of use and the age of initiation of use of: alcohol, tobacco, cannabis, amphetamines, LSD, and heroin. METHOD: Data were taken from the 1998 National Drug Strategy Household Survey, a survey of a representative sample of Australians aged 14 years and over. Nine five-year cohorts were examined among persons born between 1940 and 1984. The weighted prevalence of use by ages 15 years, 21 years, and lifetime use, was estimated, as was the average age of first use among users. The significance of trends was tested using logistic regression (for lifetime use, use by 15 and 21 years) and linear regression (for age of first use). RESULTS: Lifetime prevalence of alcohol and tobacco use was similar among all birth cohorts. The prevalence of illicit drug use--cannabis, amphetamines, LSD and heroin--increased with successive birth cohorts and more recent birth cohorts reported using licit and illicit drugs at a younger age. CONCLUSIONS: More recent cohorts are more likely to use illicit drugs at some point in their lives. Greater numbers of persons from more recent birth cohorts may be at risk of developing substance-related problems. PMID- 11011472 TI - Systematic review processes and the management of opioid withdrawal. AB - OBJECTIVE: To assess the diversity of research on the management of opioid withdrawal, identify sources of heterogeneity and provide a context for subsequent systematic reviews to establish evidence-based best practice. METHODS: References were identified through searches of multiple electronic databases and handsearching the reference lists of retrieved articles. The principal criterion for inclusion in the literature mapping process was that it be a study of an intervention intended to manage the process of opioid withdrawal. RESULTS: Of 218 references assessed, all participants were dependent on heroin in 41% and on methadone or l-alpha acetyl methadol (LAAM) in 24%. More than 17 different types of treatment approach were identified. Only 42% of references used a rating instrument to assess withdrawal severity and reported sufficient results to indicate the timing and magnitude of the peak and/or duration of withdrawal. The type of rating instrument used and the way in which results were reported varied enormously. A clear parameter for completion of detoxification was used for 37% of references. CONCLUSIONS: The capacity for rigorous systematic reviews of the management of opioid withdrawal is currently limited. There are multiple sources of heterogeneity that will need to be taken into account. IMPLICATIONS: The use of narrative reviews and observational studies are important complements to formal systematic reviews in the establishment of evidence-based practice in any area that combines aspects of psychology, behaviour, social context and medical treatment. PMID- 11011473 TI - Bilateral health aid: lessons from Europe. AB - OBJECTIVE: The objective of this study is to document current issues in policy and planning for development assistance in health from the perspective of key bilateral aid donors in Europe, and to explore its relevance for Australia. METHODOLOGY: This study is based on seventy-seven interviews and documentary analysis undertaken between July and December 1999 with policy, technical and field staff of major European bilateral aid donors, the World Health Organisation (WHO), and key academic institutions. Notes were taken of interviews, and relevant documentation collected. Forty-five key interviews were tape recorded and transcribed for analysis. RESULTS: The shift towards program and Sector Wide Approaches in development assistance reflects broader political changes in Europe since the collapse of the Communist block. Increasingly, achieving an impact on poverty is seen as linked to better governance, improved policy frameworks, and collaboration between donors and partner governments, and between donors themselves, reflected in the broad support for Sector Wide Approaches to development assistance. CONCLUSIONS: These perspectives bring new demands to donors such as Australia: structural changes, procedural changes, collaborative approaches to programs, closer relationships with counterparts. IMPLICATIONS: The uncertainty in which health sector reform operates necessitates a reflective and adaptive approach to management of aid, and responsiveness to monitoring and evaluation and the development of new knowledge. Given Australia's strategic positioning in its sphere of influence, the experience from Europe should inform the development of our own future directions. PMID- 11011474 TI - Smoking among ethnic Chinese patients and their recall of quit advice by Chinese speaking general practitioners in Sydney. AB - OBJECTIVES: To collect smoking-related data from Chinese patients attending Chinese-speaking general practitioners (CSGPs), including self-reported smoking status, recall of a question about smoking status during the previous six months by their GP and, for smokers, their recall of quit advice in their most recent consultation. DESIGN: Descriptive research first involving self-administered waiting-room questionnaires and, afterwards, a second self-administered questionnaire to examine behaviour in that most recent consultation. SETTING: Surgeries of 24 CSGPs in Central Sydney. SUBJECTS: 1084 Chinese patients aged 18 70 years. RESULTS: Self-reported smoking prevalence was 25% (95% CI: 21%-29%) for men and 4% (95% CI: 2%-5%) for women. Of 103 smokers who had visited their regular GP during the previous 6 months, 42% recalled a question about smoking status and 38% quit advice. As elicited at follow-up, 72% recalled any discussion in their most recent consultation about smoking although effective techniques were rarely recalled. Patient's sex (OR: 1.78; 95% CI: 1.28-2.47) and smoking status (OR: 1.39; 2.38; 95% CI: 0.85-2.25, 1.49-3.81) independently predicted a question about smoking status. Among smokers, marital status was the only independent predictor of quit advice. As assessed by a stage-of-change scale, significantly more Chinese smokers were not ready to contemplate cessation, compared with Angio-Celtic community survey results. CONCLUSIONS: Our study provides unique information about smoking among ethnic Chinese patients attending CSGPs. Findings also suggest CSGPs need greater support to maximize their clinical opportunities to advise Chinese smokers to quit. PMID- 11011476 TI - The turmoil of aboriginal enumeration: mobility and service population analysis in a central Australian community. AB - This paper documents Aboriginal population change and mobility over time in a remote community in central Australia. The movement of population has implications for service delivery and resource allocation. Aboriginal population in the region is characterised by high mobility. We conducted four population surveys in a selected remote community over a 12 month period and categorised individuals into four mutually exclusive groups: residents, dual residents, visitors and absent residents. Based on these categories we developed two population classifications: actual and potential service populations. The potential service population was consistently higher than the Australian Bureau of Statistics (ABS) census figure. We question the use of ABS census estimates as appropriate population figures for determining resource allocation to remote communities. We quantify inter- and intra-community mobility. When the potential population is used as denominator, 35% of the population of this community was classified as inter-community mobile. Given this level of mobility we argue that: (1) Resources should be allocated to compensate health services for the additional time and resource requirements to deal with the high level of population mobility. (2) Health programs such as STD control, trachoma, scabies and other communicable diseases common in Central Australia should be coordinated and delivered as regional programs often crossing State/Territory borders. PMID- 11011475 TI - A general practice-based recruitment strategy for colorectal cancer screening. AB - OBJECTIVE: To determine whether a screening recruitment strategy for first degree relatives of people with colorectal cancer is effective in enabling eligible relatives to request screening from their general practitioner (GP) and to assess acceptability to GPs and patients. METHODS: Thirty GPs, from 26 practices, and 303 of their patients aged over 50 who were first degree relatives of a person with colorectal cancer, participated in a randomised controlled trial of a GP based recruitment strategy, in the Newcastle Area of New South Wales, Australia. RESULTS: The proportion of relatives requesting screening was statistically significantly higher in the intervention group than in the control group (18% compared to 4%, respectively; p = 0.01). CONCLUSIONS: Interest in the study by GPs was low, however for GPs who were involved, the recruitment strategy did prompt first degree relatives to discuss screening. The strategy may be even more effective when combined with other interventions such as a media campaign. The results may be generalizable to feasibility studies of general population screening for colorectal cancer in Australia. IMPLICATIONS: The results of this work are potentially informative to public health practice in Australia given the ensuing pilot programs of colorectal cancer screening. PMID- 11011477 TI - Researching health care and public policy. PMID- 11011478 TI - Do women have equitable access to quality breast prosthesis services? AB - Although up to 90% of women who have had a mastectomy use breast prostheses, significant gaps exist around current breast prosthesis services for Australian women. These gaps include the timeliness and quality of information provision, the disparity in financial assistance, and the lack of knowledge regarding the determinants of what constitutes a "quality" breast prosthesis. Revised policy initiatives are central to addressing these gaps to ensure equitable access to quality breast prosthesis services. PMID- 11011479 TI - The long-term factors associated with removal from parents amongst indigenous prisoners in NSW. PMID- 11011480 TI - Smoke rings around the Olympics. PMID- 11011481 TI - A randomised trial of telephone versus postcard prompts to enhance response rate in a phased population-based study about community preferences. PMID- 11011482 TI - Long-term ventilating tube with tympanosclerosis. PMID- 11011483 TI - Pedunculated granuloma of the vocal fold. PMID- 11011484 TI - Endoscopic views of nasal septal polyps. PMID- 11011485 TI - Electronystagmography: dizziness and syncope. PMID- 11011486 TI - Giant-cell reparative granuloma. PMID- 11011487 TI - Are we all just stupid? PMID- 11011488 TI - Nasal congestion: a review of its etiology, evaluation, and treatment. AB - The most common clinical syndromes that cause nasal congestion are allergic rhinitis, vasomotor rhinitis, chronic sinusitis, and upper respiratory viral infections (common colds). Nasal congestion, in turn, can lead to sequelae such as sinusitis, otitis media, and the onset or worsening of mild to severe sleep disturbances, including obstructive sleep apnea. There is a host of conservative treatments, including decongestant pharmacotherapy, antiallergy measures, and nasal dilation devices. Several surgical procedures are also available. This article reviews the current guidelines for the workup and diagnosis of nasal congestion and briefly describes the many and varied approaches to treatment. PMID- 11011489 TI - Audiometric manifestations of Waardenburg's syndrome. AB - We studied 23 patients (from 11 families) who had Waardenburg's syndrome. Patients were evaluated by conventional audiometric methods and by distortion product otoacoustic emissions to determine the penetrance and the degree and type of hearing loss. Twelve of the patients had the type I form of the syndrome and 11 had type II. Overall, we found hearing loss in 19 of the 23 patients (83%); hearing loss affected nine type I patients (75%) and 10 type II patients (91%). Five type I patients (42%) and eight type II patients (73%) had a hearing loss of > 100 dB. Bilateral symmetrical hearing loss was the most common type of loss, as it was seen in six of the type I patients (50%) and eight of the type II patients (73%). At lower frequencies, distortion-product otoacoustic emission amplitudes were found to be significantly above the noise floor in five of the 11 patients whose hearing thresholds were 60 dB HL or worse by click auditory brainstem response testing. These findings led us to conclude that it is necessary to use otoacoustic emissions in patients with Waardenburg's syndrome in order to provide optimum fitting of hearing aids, especially in children. PMID- 11011490 TI - Unplanned admissions following outpatient otologic surgery: the University of Arkansas experience. AB - We analyzed the outpatient otologic surgery experience at our institution to identify those factors that are associated with a high risk of postoperative complications that require an unplanned hospital admission. We found that among a group of 662 patients who underwent group II otologic procedures (i.e., tympanoplasty with or without mastoidectomy, stapedotomy, and middle ear exploration), the overall admission rate was 4.7%, of which 3.9% were unplanned. A significantly larger percentage of children were admitted than adults (5.7 vs. 2.3%), primarily for nausea and vomiting. Three factors were significantly associated with unplanned admissions: the type of surgery (tympanomastoidectomy with ossicular reconstruction), the duration of general anesthesia (> 2 hr), and asthma as a coexisting condition. The choice of antiemetic administered (ondansetron or droperidol) and the specific agents used for general anesthesia did not appear to have any significant impact on unplanned admissions. We recommend that the three predisposing factors be taken into consideration when formulating the treatment plan. Scheduling an inpatient procedure for patients who have risk factors for complications requiring hospitalization would avoid the extra costs and stress associated with unplanned admissions. PMID- 11011491 TI - Giant benign sinonasal squamous papilloma: report of a case. AB - We treated a patient with a giant squamous papilloma in the nasal cavity and maxillary sinus that extended through a bony defect into the oral cavity. The mass was excised with a combined endoscopic, Caldwell-Luc, and transoral approach. Lesions of this type are rare, but when they do occur, the rate of recurrence is high. Therefore, long-term followup, including endoscopic examination, is important. PMID- 11011492 TI - A new ligation approach to the management of chronic epistaxis. AB - Standard cauterization therapy for chronic epistaxis is less than ideal because of the inadequacy of topical anesthesia and the difficulty of treating young, often uncooperative patients. The author has developed a new procedure for treating these patients, which entails ligation and cauterization while the patient is under light general anesthesia in an outpatient surgical facility. Of the approximately 75 procedures the author has performed, only one failed to achieve expected results. This article describes how this safe, simple, and ultimately cost-effective procedure is performed. PMID- 11011493 TI - Nasopharyngeal cyst of branchiogenic origin: report of a case and review of the literature. AB - To date, only 22 cases of nasopharyngeal cyst of branchiogenic origin have been reported in the literature. In this article, we report a new case in a 35-year old woman. We also present a review of the literature and a discussion of the sites of origin, histopathology, and treatment modalities. PMID- 11011495 TI - Primary non-Hodgkin's lymphoma of the frontal sinus: how we diagnosed it. AB - Isolated primary non-Hodgkin's lymphoma of the frontal sinus is rare. We describe the case of a middle-aged man who came to us with signs of orbital cellulitis complicating an acute infected frontal mucocele. His condition was initially controlled with medical therapy and subsequent endoscopic sinus surgery, but his symptoms eventually returned. We were able to diagnose the lymphoma only by approaching the sinus externally to obtain a biopsy. This case highlights the importance of making a full visual inspection of the involved sinus in order to avoid missing an unexpected, albeit a rarely encountered, pathology. PMID- 11011494 TI - Canalith repositioning for benign paroxysmal positional vertigo: a randomized, controlled trial. AB - We assessed the efficacy of the canalith repositioning maneuver by comparing it with no treatment in a population of patients with benign paroxysmal positional vertigo (BPPV). In this randomized, controlled, 6-month efficacy trial, outcomes were measured subjectively by patients' reports of symptom status and objectively by Hallpike testing. During the first month of the study, the treated group experienced significantly better outcomes than did the control group, but this trend was not sustained at 3 and 6 months. PMID- 11011496 TI - In memoriam: Martin T. Orne, 1927-2000. PMID- 11011498 TI - Descriptive outcomes of the American Lung Association of Ohio hypnotherapy smoking cessation program. AB - Hypnosis smoking cessation treatment is one type of program available to smokers. This paper describes a large randomly selected sample from such a program, which has not been previously reported. During 1997, 2,810 smokers participated in single-session, group hypnotherapy smoking cessation programs sponsored by the American Lung Association of Ohio. A randomly selected sample of 452 participants completed telephone interviews 5 to 15 months after attending a treatment session. Twenty-two percent of participants (n = 101) reported not smoking during the month prior to the interview. Use of other smoking cessation strategies since the treatment session were assessed. Interestingly, only 20% of participants who used pharmaceutical products to assist with smoking cessation took them for the recommended treatment duration. Hypnotherapy smoking cessation treatment offers an alternative cessation method, which may meet the unique needs of certain individuals. PMID- 11011497 TI - Specific autobiographical memory following hypnotically induced mood state. AB - This study investigated the impact of hypnotically induced mood on the specificity of autobiographical memory. High (n = 24) and low (n = 21) hypnotizable participants were administered a hypnotic induction for sad, neutral, or happy mood and were asked to retrieve specific autobiographical memories in response to positive and negative cue words. Whereas high hypnotizable participants in the sad condition provided fewer specific memories in response to positive rather than negative cues, those in the neutral and happy conditions responded similarly to positive and negative cues. Findings suggest that impaired recall of specific memories may be mediated by state factors associated with sad mood. These results point to the utility of hypnotic mood induction as a means to experimentally investigate the relationship between mood and autobiographical memory. PMID- 11011499 TI - Autonomic and psychological responses to an acute psychological stressor and relaxation: the influence of hypnotizability and absorption. AB - This study examined the influence of hypnotizability and absorption on psychological and autonomic responses to an experimental stressor and a relaxation procedure of 13 high and 13 low hypnotizable subjects. Heart-rate variability was the measure of autonomic reactivity. Absorption was found to be the only significant predictor of autonomic reactivity in both experimental conditions. Expectation and previous relaxation training, but not absorption or hypnotizability, predicted perceived relaxation in the relaxation condition. The results suggest that in a nonhypnotic context the influence of hypnotizability on responses to experimental conditions may be less prominent than the influence of absorption. Absorption may be associated with greater awareness of internal physical and psychological processes, and the results support previous clinical findings of positive correlations between absorption, subjective perception of autonomic arousal, and somatic symptom reporting. PMID- 11011500 TI - Goal-directed fantasy does not explain the training effect of the Carleton Skills Training Package. AB - Two treatments to enhance the hypnotic responsiveness of subjects who pretested as low in hypnotic susceptibility were compared. Subjects in one modification treatment (n = 20) received the Carleton Skills Training Package (CSTP). Those in a second modification treatment (n = 20) were administered a partial training that did not include information about imaginal strategies. The authors of the CSTP assume that goal-directed fantasies underlie subjects' feelings of involuntariness, so teaching imaginal strategies is the primary agent of subjective changes after the training. Controls (n = 20) received no treatment. Both training packages enhanced behavioral and subjective response to an equivalent degree. Control subjects' performances were stable across tests. Modest increases in hypnotizability scores following training were related neither to enhancing goal-directed fantasies nor to use of imagery. Other mechanisms, possibly compliance, may underlie the CSTP effect. PMID- 11011501 TI - Ericksonian approaches to hypnosis and therapy: where are we now? AB - Ericksonian approaches to psychotherapy and hypnosis have had a significant impact on many clinical practitioners over the last two decades. This article reviews the current empirical research with regard to the efficacy of these treatment approaches as well as for the key Ericksonian assumptions of: (a) belief in an altered state of consciousness and the existence of specific markers indicating an altered state; (b) the superiority of indirect suggestion over direct suggestion; and (c) client hypnotizability is a function of the hypnotist's skill. The current literature provides empirical support neither for efficacy nor for these key assumptions. The article concludes with a discussion of the need for empirically based research to test the efficacy of Ericksonian therapy and its core components, lest this approach become isolated from the scientific hypnosis and therapy communities. PMID- 11011502 TI - Where Ericksonian legend meets scientific method: a comment on Matthews. PMID- 11011503 TI - The effect of union type on psychological well-being: depression among cohabitors versus marrieds. AB - Marital status is a key determinant of psychological well-being. I use data from both waves of the National Survey of Families and Households to evaluate the effect of union type (i.e., cohabitation versus marriage) on depression. Cohabitors report higher levels of depression than their married counterparts, net of sociodemographic factors. The greater depression characterizing cohabitors is primarily due to their higher relationship instability relative to marrieds. Cohabitors' reports of relationship instability are about 25 percent higher than marrieds' reports. High levels of relationship instability are especially detrimental for cohabitors who have been in their union for a long period of time. Additionally, cohabitors' depression scores are exacerbated by the presence of biological and step children, whereas marrieds' depression scores are impervious to children. Longitudinal analyses that correct for selection bias confirm that the lower levels of well-being characterizing cohabitors are not due to the types of people who choose to cohabit. PMID- 11011504 TI - Life stress, anger and anxiety, and delinquency: an empirical test of general strain theory. AB - General strain theory (Agnew 1992) departs from traditional strain theories by emphasizing the role of the individual's affective responses to negative life experiences in fostering deviant behavior. In this analysis, we examine the central hypotheses of general strain theory using data from a three-wave panel study of high school youths in the Boston metropolitan area (N = 939). Covariance structure models reveal that anger and hostility in response to negative life events do play a causal role in fostering more aggressive forms of delinquency, but are not significantly related to either nonaggressive delinquency or marijuana use. Furthermore, the conditional effects predicted by general strain theory, in which the impact of strain on delinquency varies by youths' personal and social resources, are inconsistent. Discussion centers on the prospect of increasing the utility of general strain theory by further imbuing it with concepts and perspectives from the sociology of mental illness. PMID- 11011505 TI - Alcohol and employment in the transition to adulthood. AB - We elaborate the relationship between work hours and alcohol use during the transition from adolescence to adulthood. Both hours of employment and drinking may be products of weak bonds to school and family. Alternatively, work may exert an independent effect on alcohol use by exposing adolescents to opportunities and associates that facilitate drinking. Using longitudinal data from the Youth Development Study (YDS), we present static score regression models showing that long work hours increase levels of drinking during high school. These effects are mediated in large part by work-derived independence from parents, suggesting that a precocious transition to adult roles may be the mechanism connecting work hours and alcohol use. Work effects on drinking are short-lived, however, as adolescent hours of employment do not significantly influence alcohol use after high school. PMID- 11011506 TI - Perceived discrimination and depression among Mexican-origin adults in California. AB - We contend that perceived discrimination has an independent effect on depression outcomes among adults of Mexican origin. Using a sample of 3,012 Mexican-origin respondents in Fresno, California in 1995/96 (ages 18-59) we investigate the direct and moderating connections between perceived discrimination, acculturative stress, and mental health (CES-D). We also investigate the social patterning of perceived discrimination. While more highly acculturated immigrant respondents were more likely to experience discrimination than their less acculturated counterparts, more highly acculturated U.S. born respondents were less likely to experience discrimination. Discrimination was directly related to depression, but this effect was moderated through nativity/country of residence, English-language acculturation, sex, and country of education variables. Moderate levels of legal status acculturative stress were especially depressive for native-born U.S. residents. PMID- 11011508 TI - Settlement has many faces: physicians, attorneys and medical malpractice. AB - We conduct an analysis of the jurisdictional dispute over the management of medical malpractice lawsuits, focusing on the process through which liability is defined. We utilize a North Carolina sample of physicians who have been sued, their defense counsel, and counsel for the plaintiff in the case. A comparison of the perspectives of these three parties reveals that over half of the physicians who settle perceive themselves as not liable. Defense counsel are more adept at predicting both negotiated resolutions and whether or not money will be paid than either plaintiffs' counsel or physicians. Almost two-thirds of physicians who thought they were not liable expressed a desire for vindication. Almost half the time when the physicians denied liability money was nonetheless paid to resolve the claim. Physician responses to the outcome of their cases focus on the need for reform, especially in terms of a call for peer or expert review. We identify and discuss culture conflict between law and medicine. For lawyers "settlement" is not a negative thing, but for physicians it implies fault. We challenge existing literature which analyzes the settlement of medical malpractice claims solely in terms of rational economic models, and we argue that social psychological variables are equally important. PMID- 11011507 TI - Unfair treatment, neighborhood effects, and mental health in the Detroit metropolitan area. AB - Why do racial differences in many indicators of mental and emotional well-being show inconsistent patterns? We propose that mental and emotional well-being are influenced by aspects of the social context, including experiences of unfair treatment and the concentration of households with incomes below the poverty level, and that differential exposure to these factors influences racial differences in mental well-being. We analyze the reporting of psychological distress and life satisfaction in a multistage area probability sample of 1,139 African American and white residents of the Detroit metropolitan area aged 18 and older. Both psychological distress and life satisfaction are significantly associated with exposure to unfair treatment and with the proportion of households in the census block group that were below the poverty level. Racial differences in psychological distress and life satisfaction were eliminated or reversed once differentials in the percent of households living below the poverty line and exposure to unfair treatment were accounted for. These findings contribute to a growing body of evidence that "race" effects operate through multiple pathways that include race-based residential segregation and its attendant economic disinvestment at the community level, and interpersonal experiences of unfair treatment. PMID- 11011509 TI - Death makes news: the social impact of disease on newspaper coverage. AB - This paper is an integrated analysis of newspaper coverage, epidemiological rates, and recent social history of six prominent diseases. HYPOTHESES: Newspaper coverage of a disease has three developmental stages (emergence, maturation, and decline & death). Trends in newspaper coverage of a disease reflect trends in its mortality, prevalence, and incidence. Magnitudes of newspaper coverage of diseases reflect their differential mortality rates. DATA: Using the LEXIS-NEXIS news archive for major U.S. newspapers, we retrieve articles about cancer, heart disease, AIDS, diabetes, Alzheimer disease, and arthritis for the period 1977 1997. We also obtain mortality, prevalence, and incidence trends for the six diseases. RESULTS: During the two decades, newspaper coverage emerges for AIDS and Alzheimer disease and is in the mature stage for the other diseases; declines begin for heart disease and AIDS. Trends in news coverage closely parallel mortality trends, and less consistently prevalence and incidence trends. Sharp downturns and upturns in mortality are mirrored in news volume. High-mortality diseases prompt both the most news coverage and the largest proportions of articles with death topics. CONCLUSION: Newspaper coverage of diseases is responsive to their mortality levels and trends. PMID- 11011510 TI - Homophone meaning generation: a new test of verbal switching for the detection of frontal lobe dysfunction. AB - Executive functions in their broadest sense may be impaired in patients with frontal lobe lesions. The Wisconsin Card Sorting Test, perhaps the most robust test sensitive to frontal lobe dysfunction, requires the flexibility to switch a response pattern to meet the change in task demands. However this task has only two category switches and normal respondents tend to score at ceiling. Verbal fluency tasks also incorporate a switching component and it was hypothesized that a word generation task that maximized a switching requirement might provide a more satisfactory verbal measure of frontal lobe dysfunction. A new test of homophone meaning generation that requires multiple switches between verbal concepts (e.g., tick = insect, correct, etc.) was devised. Normative data was obtained from a sample of 170 control participants. Seventy-one patients with unilateral anterior or posterior cerebral lesions were also tested. A normal distribution of scores was obtained in the standardization sample. The anterior lesion groups were more impaired than the posterior groups. There were no significant differences due to laterality. This homophone meaning generation task is a measure of frontal lobe dysfunction that has the advantage of psychometric properties that permit measurement of the degree of impairment. PMID- 11011513 TI - Does literacy have an effect on stick construction tasks? AB - Since constructional apraxia is often concomitant with brain lesions, the study of constructional tasks in the non-brain-damaged population might be useful in helping to disentangle other causal factors. This paper explores the performance of illiterate individuals (N = 29) as compared to that of semiliterates (N = 21) and literates (N = 23) in order to see the effect of reading and writing abilities on constructional tasks. Each participant was asked to construct 4 figures based upon models having varying degrees of complexity. A global criterion of lack of fidelity and several analytic criteria (related to distortion, rotation, and disarticulation errors) were used to evaluate performance. Although illiterates generally made more errors than semiliterates and semiliterates more than literates, only some of these differences were statistically significant. Significant differences were found for lack of global fidelity and disarticulation errors when all 4 figures were considered together. Subtler data emerged with respect to single figures. PMID- 11011511 TI - The functional relevance of affect recognition errors in schizophrenia. AB - To evaluate the clinical and ecological validity of affect recognition (AR) measures in a sample of community-dwelling schizophrenic outpatients (N = 40), we analyzed the relation of facial and vocal AR to intellectual, symptomatic, and quality-of-life criteria. Facial and vocal AR showed virtually identical patterns of association with these criteria, suggesting that both modalities of AR draw on the same underlying heteromodal capacity. Specifically, AR was correlated with a subset of intellectual abilities (verbal-semantic, executive-attentional), but was unrelated to age, education, or neuroleptic dose. In terms of clinical and ecological criteria, AR errors correlated with more severe psychotic symptoms (positive and disorganized) and with lower quality of life (relationships, community participation, and richness of intrapsychic experience). Even after controlling for subjects' intellectual abilities and illness severity, inaccurate AR was associated with bizarre behaviors (involving sociosexual interactions, clothing, appearance) and with impoverished interpersonal relations. Thus, while difficulty identifying basic affective cues is related to general cognitive and illness-severity factors, it appears to have specific functional implications that do not depend on generalized impairment. Assessment of AR may identify a subgroup of schizophrenic patients who have a central defect in the heteromodal monitoring of emotional-social displays, associated with dysregulation of social behaviors and disruption of interpersonal relations. PMID- 11011512 TI - Affective self-report during the intracarotid sodium amobarbital test: group differences. AB - Emotional reactions are sometimes observed during the intracarotid sodium amobarbital test. For instance, euphoric/indifference reactions can be seen during right hemisphere inactivation and catastrophic reactions may accompany left hemisphere inactivation. Less dramatic changes can also be detected in affective self-report during left and right hemisphere amobarbital tests, with more negative affect reported during left hemisphere inactivation and either neutral or mildly positive affective states reported during right hemisphere inactivation. The current study not only replicated this effect, but in addition, found significant group differences. The first group (right way) showed a pattern of affective self-report during left and right amobarbital tests entirely consistent with prior findings, while a second group (wrong way) showed results that behaved in a diametrically opposite fashion. A third group (no change) showed little, if any, difference in affective self-report during left and right amobarbital tests. The major factor distinguishing the wrong way group from the other two appeared to be an asymmetrical distribution of left and right temporal lobe lesions in the former group. In contrast, the factor differentiating the right way group from the no change group appeared to be the relative degree of left hemisphere inactivation during the left hemisphere amobarbital test. The results are discussed not only in terms of their impact on theories of cerebral lateralization for emotion, but also in terms of methodological issues in this field. PMID- 11011514 TI - Characteristics of memory dysfunction in body dysmorphic disorder. AB - Although body dysmorphic disorder (BDD) is receiving increasing empirical attention, very little is known about neuropsychological deficits in this disorder. The current study investigated the nature of memory dysfunction in BDD, including the relationship between encoding strategies and verbal and nonverbal memory performance. We evaluated 17 patients with BDD and 17 healthy controls using the Rey-Osterrieth Complex Figure Test (RCFT) and the California Verbal Learning Test (CVLT). BDD patients differed significantly from healthy controls on verbal and nonverbal learning and memory indices. Multiple regression analyses revealed that group differences in free recall were statistically mediated by deficits in organizational strategies in the BDD cohort. These findings are similar to patterns previously observed in obsessive-compulsive disorder (OCD), suggesting a potential relationship between OCD and BDD. Studies in both groups have shown that verbal and nonverbal memory deficits are affected by impaired strategic processing. PMID- 11011515 TI - Perception and estimation of time in long-term survivors of childhood posterior fossa tumors. AB - We examined short duration perception (400 ms), long duration estimation (30 and 60 min), and spatiotemporal estimation in long-term survivors of childhood cerebellar tumors with a mean time since diagnosis of 14.2 years. Groups of individuals with tumors treated with surgery only (astrocytoma, N = 20) were compared to those with tumors treated with surgery, focal radiation, and craniospinal radiation (medulloblastoma, N = 20), and to age-matched controls (N = 40). Childhood lesions of the cerebellum produced enduring deficits in short duration perception, but spared the ability to functionally estimate long durations, regardless of the pathology or treatment of the tumor. Evidence did not support any functional recovery over time of the cerebellar system that underlies short-duration perception. Younger age at treatment was not a protective factor. Although no group differences were present in the functional measures of long-duration estimation, tumor-related prospective memory deficits interfered with the ability to produce long-duration prospective estimates. The utilization of sensory and somatomotor information to refine real-world spatiotemporal estimates was compromised in the medulloblastoma group only. PMID- 11011516 TI - Conceptual apraxia and semantic memory deficit in Alzheimer's disease: two sides of the same coin? AB - This study was designed to examine the patterns of apraxic disturbances and the relationships between action knowledge and other measures of semantic knowledge about objects in 10 well-characterized Alzheimer's disease (AD) patients. Five tasks were used to assess components of action knowledge (action-tool relationships, pantomime recognition, and sequential organization of action) and praxis execution (actual use, pantomiming) according to the cognitive model of praxis. Three tasks (verbal comprehension, naming, and a visual semantic matching task) were used to assess verbal-visual semantics. Considering patterns of apraxia first, conceptual apraxia was found in 9 out of the 10 AD patients, suggesting that it is a common feature even in the early stages of AD. Second, we found partly parallel deficits in tests of action-semantic and verbal-visual semantic knowledge in 9 AD patients. Impaired action knowledge was found only in patients with a semantic language deficit. These findings provide no evidence that "action semantics" may be separated from other semantic information. Our results support the view of a unitary semantic system, given that the representations of action-semantic and other semantic knowledge of objects are often simultaneously disrupted in AD. PMID- 11011518 TI - In defense of the frontal lobe hypothesis of cognitive aging. PMID- 11011517 TI - The frontal aging hypothesis evaluated. AB - That the human frontal lobes are particularly vulnerable to age-related deterioration has been frequently invoked as an explanation of functional decline in aging. This "frontal aging hypothesis" is evaluated in this review by examining evidence of selectively reduced frontal lobe function in aging. The frontal aging hypothesis predicts that functions largely dependent on frontal regions would decline in aging, while functions largely independent of frontal lobes would remain relatively spared. The hypothesis further predicts that age related brain change would selectively impact frontal regions. The literatures on working memory, visuospatial attention, face recognition, and implicit memory were reviewed as exemplars of functions dependent on prefrontal, parietal, temporal and occipitotemporal cortices, respectively, with a view to establishing mediating structures and effects of aging. Age sensitivity was seen both in functions dependent on frontal integrity as well as in functions apparently independent of frontal integrity. Further, although prefrontal areas exhibit age related decreases in regional volume, blood flow and metabolism, nonfrontal cortical regions undergo similar declines. It is concluded that while the frontal lobes are subject to age-related changes reflected in both behavior and pathology, there is only weak and conflicting evidence that frontal regions are selectively and differentially affected by aging. It is argued that a network based theory of cognitive aging has advantages over the localizationist approach inherent in the frontal aging hypothesis. PMID- 11011519 TI - Negative seizures. AB - Negative phenomena can occur with seizures, but some ictal negative manifestations are rare and may lead to misdiagnosis. A patient series is presented with unusual ictal negative phenomena: neglect syndrome, catastrophic depression, apraxia, aphasia, amnesia, homonomous hemianopsia, and hemiparesis. One had repeated episodes with PLEDs but no parenchymal lesions. Clinicians should consider seizures in the setting of unexplained deficits, even if there are no positive ictal phenomena. PMID- 11011520 TI - ECG of the month. The untamed heart. Supraventricular tachycardia, type? PMID- 11011521 TI - Surgical management of the aging upper face. AB - Creating a more youthful appearance of the aging upper face requires a treatment plan which is tailored to address the patient's individual needs. The process begins with a comprehensive knowledge of anatomy and the physical effects of aging, involves discussion and understanding of aesthetic ideals, and culminates in the selection and performance of the appropriate procedure. PMID- 11011522 TI - Radiology case of the month. Incidental discovery on mammography done for a palpable breast mass. Granular cell tumor. AB - A 32-two-year-old woman complains of a palpable lump in her left breast. Her past medical and family history is non-contributory. There was no palpable lump on clinical breast examination. A mammogram was performed. PMID- 11011523 TI - A clinical report on intravenous saline infusion in the wards of the New Orleans Charity Hospital from June 1888 to June 1891. PMID- 11011524 TI - The Medical Education Commission report at the turn of then new millennium 2000. AB - The State of Louisiana Medical Education Commission was formed by Act 3 of the 1997 Louisiana Legislature. The members are appointed by the Louisiana State University Health Sciences Center, Tulane University Medical Center, and Alton Ochsner Medical Foundation and report to and advise the Department of Health and Hospitals (DHH). This summation from the Medical Education Committee is designed to answer three questions. First, how does the State of Louisiana compare nationally in the renewal of physician supply and capability? Second, what are the current status and trends of graduate medical education in Louisiana? Third, what recommendations are proposed to continue and improve the important relationship between the State public hospitals and educational programs? The State of Louisiana has participated proportionately relative to population in the growth and progress of medical education in this last century and compares favorably with other states and the nation. Louisiana exceeds national averages in the increase of primary care residency programs and positions and in the retention of trainees in practice sites in the State. The three-year trends in total number of graduate medical education filled positions has been consistent with only 0.4% change, with primary care showing an overall increase of 9%, reflecting increases in Family Medicine (56%) and Medicine/Pediatrics (41%). PMID- 11011525 TI - Does the admissions committee select medical students in its own image? AB - A previous issue of the Journal referred to a national study of the Myers-Briggs Type Indicator (MBTI) in which 12 schools participated. A comparison of our students' type preference with that of their peers at the 11 other schools revealed that only two of the 11 schools demonstrated our strong preference for the combination of sensing, thinking, and judging. We wondered if our Admissions Committee members, too, would demonstrate a preference for the same combination of Myers-Briggs dimensions. In fact, students' trait preferences matched those of Admissions Committee members on all MBTI dichotomies, suggesting that the Committee's own image may have influenced the selection of medical students. A Committee member's awareness and appreciation of his own personality characteristics may be extremely helpful in the admissions process. PMID- 11011527 TI - Kind strangers? Physicians through the eyes of Tennessee Williams. AB - In Louisiana, Tennessee Williams is usually thought of as a famous denizen of the French Quarter or perhaps as our greatest playwright. Medicine rarely enters into it. Illness, however, particularly mental illness, shaped much of Williams' life and his work. The playwright had mixed feelings about physicians and their effect on his life and that of his close relations. These feelings worked their way into his plays. Through it all Williams gives a vivid, humorous, and deeply truthful image of the doctor-patient relationship in the first half of the twentieth century. Here we give a brief review of medicine in Williams' work. PMID- 11011526 TI - Tuberculosis screening, referral, and treatment in an inner city homeless shelter in Orleans parish. AB - Tuberculosis screening and preventive therapy among the homeless has been a challenge because of the lack of coordinated follow-up. Homeless persons at a homeless shelter in inner city New Orleans were screened for tuberculosis infection and referred for follow-up evaluation and preventive therapy. Fifty-two percent of the 104 persons screened completed the initial evaluation. Twenty-two percent of these patients had latent tuberculosis infection. Forty-two percent of infected patients completed the referral and follow-up process. Patients during the second 3 months of the program were twice as likely to complete the initial evaluation, the referral, and the follow-up process as were patients during the first 3 months due to enhanced awareness and increased educational intervention. A competent referral system for homeless persons may be achieved by implementing a single-clinic, on-site tuberculosis screening and follow-up system with the active participation and coordination of state agencies, the medical community, and organizations which operate homeless facilities. PMID- 11011528 TI - Six degrees of Kevin Bacon--Al Eskan disease and "dirty dust.". PMID- 11011529 TI - Lower Marine combat mortality. PMID- 11011530 TI - Lumbar puncture headache exacerbated by recumbent position. PMID- 11011531 TI - Vietnam nursing: the experience lives on. Interview by Margaret Ann La Salle. AB - Through personal interviews and a review of the literature, this article examines the experiences of three nurses who served in the Vietnam conflict. Common themes emerging were the strength of the medical teams, the camaraderie, and the willingness of everyone to pitch in. Common coping strategies were turning to God for comfort and emotional support through peers. PMID- 11011533 TI - Psychological changes of Japan Self-Defense Forces personnel during selection and training for the peacekeeping mission in the Golan Heights. AB - We sought to determine whether a selection process for deployment had a measurable effect on psychological symptoms by comparing scores on the Yatabe Guilford Personality Index, the Manifest Anxiety Scale, and the 30-item version of the General Health Questionnaire between deployed and nondeployed mission candidates from the Japan Self-Defense Forces. The studies were undertaken in Japan during education and training for the United Nations peacekeeping mission in the Golan Heights. The participants included 80 candidates for deployment. Personnel who were not deployed had significantly higher measures of manifest anxiety and general psychological distress than deployed personnel, whereas deployed personnel showed more symptoms suggesting somatization. The selection process and training for deployment appear to have been stressful for all personnel, whether deployed or not. PMID- 11011532 TI - Injuries and injury risk factors among men and women in U.S. Army Combat Medic Advanced individual training. AB - No previous reports have evaluated injuries or injury risk factors during the advanced individual training (AIT) that follows the Army's initial or basic combat training (BCT). This study examined injuries and injury risk factors among 439 men and 287 women participating in combat medic AIT. A questionnaire addressing demographic and lifestyle characteristics (age, race, tobacco and alcohol use, physical activity, etc.) was administered to all subjects. Stature and body mass were obtained from battalion records. Injuries occurring during both BCT and AIT were transcribed from subject medical records. Results indicated that cumulative injury incidence (subjects with one or more injuries) in BCT was 26% for men and 52% for women (p < 0.01), in consonance with previous investigations. In AIT, injury incidence was 24% for men and 30% for women (p = 0.08). In both BCT and AIT, overuse injuries and lower body injuries accounted for the largest proportions of injuries by diagnosis and anatomical location. Logistic regression revealed that older age (> 25 years), split option (a break in service between BCT and AIT), and higher body mass were independent risk factors for AIT injuries among women. None of the examined variables were independent risk factors for AIT injuries among men. PMID- 11011534 TI - Personality characteristics of South African Navy submarine personnel. AB - This study aimed to determine the extent to which the Institute of Personality and Ability Testing Anxiety Scale and the 16 Personality Factor Questionnaire can be used to describe successful submariners in the South African Navy. The Anxiety Scale scores were within normal limits when the profiles of 85 submariners were analyzed to describe the personality characteristics of submarine personnel. Although no significant factors were found, the results indicate that four personality factors appeared to be most descriptive of the sample. They were adventurousness, confidence, group orientation, and self-sentiment. These factors appeared appropriate when discussed from an environmental demand perspective. PMID- 11011535 TI - A field-expedient algorithmic approach to the clinical management of chemical and biological casualties. AB - Warriors on the modern battlefield face considerable danger from possible attack with chemical and biological weapons. Aggravating this danger is the fact that medical resources at the lowest echelons of care, already likely to be strained to capacity during modern conventional combat, are at present inadequate to handle large numbers of chemical or biological casualties. Complicating this problem further is the austere nature of diagnostic modalities available at lower echelons. With this in mind, and given the urgency required to adequately manage chemical and biological casualties, it is likely that such casualties will initially require significant empiric care in the absence of a definitive diagnosis. Such care under field conditions, often rendered by relatively inexperienced medical personnel, might best be provided using an algorithmic approach. We have developed such an algorithm. PMID- 11011536 TI - Physiological and practical evaluation of a biological/chemical protective device for infants. AB - The Chemical Infant Protective System (CHIPS) is a special hood-like system into which a small battery-operated blower delivers filtered air. Because it is a semiclosed system, there is a risk of dangerous CO2 accumulation within the device, which particularly affects infants with acute or chronic respiratory disorders. Eleven infants hospitalized with various respiratory illnesses wore the device for 15 minutes. Inspired O2, inspired CO2, heart rate, respiratory rate, oxygen saturation, and inside temperature and humidity were measured before and during this test period. Inspired O2 and heart rate during the test period were significantly lower than baseline levels (O2, 19.1 vs. 20.1%; heart rate, 133 vs. 142 beats/min). Inspired CO2 and inside temperature during the test period were significantly higher than baseline levels (CO2, 0.23 vs. 0.06%; temperature, 25.0 vs. 23.1 degrees C). Oxygen saturation, respiratory rate, and humidity were not different from baseline levels. A short-term stay within the CHIPS in well-ventilated surroundings did not result in significant clinical and physiological impact for sick infants. Nevertheless, trends were identified that may be worrisome during longer periods and in sealed rooms. PMID- 11011537 TI - The utility of screening chest radiographs for flight physicals. AB - Previous studies have shown low efficacy of screening chest radiographs in various populations. Findings of approximately 3,500 screening chest radiographs performed for flight duty were reviewed to determine the rate of detection of significant abnormalities. There were abnormal findings in 107 chest radiographs (3%). Fifty-five of these (1.6% of total), after additional evaluation, were found to be false positive. Only two medically significant conditions were found in the screening population. Based on our data, routine screening of flight duty applicants does not appear to be justified. PMID- 11011538 TI - Surgical implications of multicentricity in renal cell carcinoma. AB - PURPOSE: To determine the incidence and metastatic pattern of multifocal renal cell carcinoma in radical nephrectomy specimens and to use these findings to guide the choice of operative procedure. MATERIALS AND METHODS: Pathology reports for 161 patients who underwent radical nephrectomy at Walter Reed Army Medical Center between 1980 and 1995 were reviewed to evaluate tumor size, stage, and incidence of multifocal tumors. RESULTS: Multicentric neoplasms occurred in 16 of 161 (9.9%) radical nephrectomy specimens from the same institution. Thirteen of the 16 multifocal tumors had cross-sectional diameters > or = 3.5 cm. Of the 16 multifocal carcinomas, 11 were noted in stage I tumors. Multifocal neoplasms were not identified with primary tumors of < 2 cm. CONCLUSION: Our data support previous reports that the incidence of multicentric renal malignancy is low and suggest that patients with small, unilateral masses (< 2 cm) can undergo kidney sparing surgery with little risk of local recurrence. PMID- 11011539 TI - Diagnoses, demographics, and utilization of care as encountered by three U.S. Navy general medical officers. AB - U.S. Navy general medical officers (GMOs) are physicians serving as general practitioners. Although exceptions exist, most GMOs are not board-certified in a specialty. They are post-graduate year 1 (PGY-1)-trained, state-licensed physicians analogous to civilian general practitioners. We conducted a retrospective study using data generated from patient visits with active duty males and females from June 1 to 30, 1998, to describe diagnoses, demographics, and utilization of care patterns encountered by three PGY-1-trained GMOs at an ambulatory clinic. A total of 781 patient encounters with 123 diagnoses from a patient population of 3,178 were recorded. This is an average of 260 patient encounters per GMO, at a rate of 2.52 patients seen per patient-care hour. Fifty seven consultations/referrals were requested (7.3% of encounters, 1.8% of the patient population). Personnel assigned to the clinic accounted for 4.2% of visits (2% of the patient population). Patient satisfaction was rated as "excellent" to "satisfactory," and no significant morbidity was observed at 1.5 year follow-up. With PGY-1 training, GMOs provide primary care to a substantial volume of prescreened patients and treat patients with a majority of diagnoses without referral or unacceptable complications. The role of GMOs, and perhaps other physicians without specialty training (i.e., general practitioners), in selected settings seems valid and may have advantageous medicoeconomic implications for military and civilian managed care systems. PMID- 11011540 TI - Noise attenuation of hearing protectors against heavy weapon noise. AB - This study evaluated the noise attenuation of earplugs and earmuffs or their combined use against heavy weapon noise in field conditions for military personnel. The noise attenuation was measured with a miniature microphone inserted into the ear canal. The subjects (13) were tested against pink noise and against the noise of explosions and bazooka, mortar, cannon, and howitzer. The attenuation (insertion loss) was 16 to 23 dB for earplugs, 10 to 20 dB for earmuffs, and 24 to 34 dB for the combined use of plugs and muffs. The transfer function of an open ear was 5 to 7 dB when measured as the C-weighted peak level. The combined use of earplugs and earmuffs gave smaller attenuation values than expected. If the limit for the C-weighted peak level is 140 dB for unprotected ears, then protection against low-frequency noise is provided for up to 156 dB by earplugs, up to 150 dB by earmuffs, and up to 165 dB by the combined use of plugs and muffs. PMID- 11011541 TI - Eye injuries in the U.S. Armed Forces. AB - OBJECTIVE: To develop prevention and treatment modalities for eye injuries, ophthalmologists require epidemiological data on the various types of eye injuries. This study sought to define eye injury patterns in the U.S. armed forces. METHOD: Data on patterns of eye injury in the armed forces were obtained through voluntary reporting by U.S. military ophthalmologists throughout the world. The reporting format was standardized with the U.S. Eye Injury Registry initial and follow-up report forms. The data were analyzed for significant injury patterns. RESULTS: Data on 112 patients were submitted, representing a broad range of the military population. Data on a total of 96 patients with a 6-month follow-up were analyzed in this study. Immediately after injury, 43% of the patients were noted to have poor vision (worse than 20/200). After treatment, only 20% were noted to have poor vision. Patients lost an average of 21.6 days of work after a severe eye injury. CONCLUSION: An eye injury is a traumatic and potentially debilitating event. The loss of visual acuity can be drastic, resulting in an extensive recovery period. PMID- 11011542 TI - Care for the Caregivers: a program for Canadian military chaplains after serving in NATO and United Nations peacekeeping missions in the 1990s. AB - The Mental Health Department of the Canadian Forces Support Unit (Ottawa) developed the Care for the Caregivers program to help participants deal with stressful events experienced directly or vicariously from the NATO and United Nations military missions of the 1990s. The program was developed after complaints of postdeployment stress were received from various military care providers. The objectives were to improve the skills of support personnel and to reduce the distress that some caregivers experienced. Thirty-one chaplains who had been exposed to stressful military operations participated in five workshops. These educational 4-day small-group workshops covered topics such as post traumatic stress disorder, vicarious traumatization, coping techniques, spirituality, self-care, and family issues. An adult education model was chosen to encourage dialogue. Outcomes included reports of professional and personal benefits, requests for additional programs, local education initiatives, and referrals to mental health professionals. Having met its objectives, the program has become a normal concluding part of stressful deployments. PMID- 11011543 TI - A model of the expected occurrence of adverse pregnancy outcomes aboard U.S. Navy ships. AB - The goal of this study was to model the incidence of ectopic pregnancy and spontaneous abortion if pregnant women in the first 20 weeks of gestation were to remain aboard ship while at sea during deployments. Ectopic pregnancies and other pregnancy complications at sea can be life-threatening events. Data sources included shipboard medical departments, an Enlisted Personnel Survey, and the Naval Health Research Center Hospitalized Pregnancy and Women Aboard Ship studies. The overall pregnancy rate was 19 per 100 woman-years (95% confidence interval, 18-20), based on the complement of women assigned to participating ships. If pregnant women routinely were to remain aboard ships at sea during deployments through their first 20 weeks of pregnancy, it is expected that approximately 9 ectopic pregnancies and 40 spontaneous abortions would occur aboard ships at sea. PMID- 11011544 TI - Efficacy of 1% permethrin for the treatment of head louse infestations among Kosovar refugees. AB - We assessed the prevalence of head louse infestation and the effectiveness of 1% permethrin against head lice in Kosovar refugees. A currently infested case was defined as a person with observable crawling lice (adults or nymphs) or a person with nits on the hair shaft within a quarter-inch of the scalp. Of the 1,051 refugees screened upon arrival in the United States, 107 (10%) were infested. Crawling lice (adults or nymphs) were observed on 62 (6%) of the individuals examined. Refugees with crawling lice were treated with a pediculicide containing 1% permethrin. Of these, 57 were reexamined the next day. Twenty of the 57 individuals were reexamined 7 days after treatment. No crawling lice were found on any of the refugees examined after treatment. We conclude that 1% permethrin treatment was effective in louse control in this refugee population. PMID- 11011545 TI - Comparison of service member and military spouse satisfaction with installation fitness facilities and exercise programs. AB - Satisfaction with fitness facilities has long been accepted as a positive contributing factor to physical activity, readiness, and overall quality of life for military families. Our findings are based on a random sample of military families surveyed at 38 installations worldwide and at remote locations. A total of 8,572 service member and 3,493 spouse (55% and 32% response rate, respectively) questionnaires were completed and returned. Overall, members were satisfied with fitness facilities and programs, but spouses were less satisfied and more unfamiliar with these facilities. Most valued programs were fitness centers and swimming pools. Members reported that elimination of fitness facilities would greatly decrease the quality of life on installations, particularly at sites outside the continental United States, whereas spouses reported that this would not have a dramatic effect. The results of this study could be used to direct funding allocations, improve the health and fitness of military families, and suggest areas for further research. PMID- 11011546 TI - Trust, efficiency, and cooperation the heart of improving the quality of health care in New Jersey. PMID- 11011547 TI - HMOs still avoid meaningful patient care reform. PMID- 11011548 TI - Senate candidates answer health care questions. PMID- 11011549 TI - The prompt-pay law and other issues. PMID- 11011551 TI - Anthrax. An old disease returns as a bioterrorism weapon. PMID- 11011550 TI - With all deliberate speed: results of the first New Jersey Physician Prompt-Pay Survey. PMID- 11011552 TI - Avoiding the hostile work environment. PMID- 11011553 TI - Umbilical cord blood stem cell transplantation and banking. PMID- 11011554 TI - A provider's risk management services checklist. PMID- 11011555 TI - A mighty task. Remarks on the swearing-in of Paul W. Armstrong as Superior Court Judge. PMID- 11011556 TI - The irony that follows Polly Murray's good work. PMID- 11011557 TI - Doctor Kitzhaber/Governor Kitzhaber. PMID- 11011558 TI - On being a scientist. PMID- 11011559 TI - The Tao of medicine. PMID- 11011560 TI - The Alfred Blalock centennial. PMID- 11011561 TI - Lessons of syphilis for the age of AIDS. PMID- 11011562 TI - Return to Aesop's fables. PMID- 11011563 TI - Birthright. PMID- 11011565 TI - Professionalism. PMID- 11011566 TI - Abortion. PMID- 11011564 TI - Audivisual memoirs of leaders in American medicine. Gertrude B. Elion, D.Sc. (Hon.), 1918-1999. PMID- 11011567 TI - Eavesdropping on the womb: the advent of fetal auscultation. PMID- 11011568 TI - The quintuple threat. PMID- 11011569 TI - "The right to die mad". PMID- 11011570 TI - [On the necessity to use better diagnostic methods and treatment of ENT diseases]. PMID- 11011571 TI - [Dysfunction of eustachian tube. New aspects of diagnostics and treatment]. AB - The authors review and provide a comparative assessment of diagnostic and treatment methods in acoustic tube dysfunction. New etiopathologically sound methods of treating long-term acoustic tube dysfunction are proposed. Therapeutic results are analysed for 36 patients with tuber dysfunction. PMID- 11011572 TI - [Potentials for the endonasal endoscopic surgery in the treatment of maxillary cysts]. AB - To evaluate efficacy of endoscopic surgery in the treatment of maxillary cysts (MC), 152 operations in 134 patients were analysed. Besides MC, 91% of the patients had additional involvement of the nasal cavity and middle nasal passage which required surgery. By computer tomography data, all the cysts were divided into four groups by wall location: posteroinferior, superior, lateral, medial or anterior. Such division is important for the choice of the surgical approach: cysts of the first three locations can be easily removed through the middle nasal passage, in medial and anterior cysts this approach is effective only in half the cases. In the rest cases the sinus was opened through the anterior wall. In all, removal of the cyst by the endonasal approach proved feasible in 73.7%, by the anterior wall--in 26.3% of cases. The algorithm proposed for the choice of optimal intervention implies the initial use of the external approach only in MC located on the anterior and medial walls in the absence of concomitant lesions. In all the other cases, surgery should begin with partial resection of the unciform process and revision of the natural anastomosis of the sinus. PMID- 11011573 TI - [Contemporary status of the issues in the treatment of children with respiratory papillomatosis]. AB - For 15 years the authors treated 205 children aged 9 months to 15 years with respiratory papillomatosis. Besides surgical endolaryngeal removal of the papillomas, they applied immunomodulators, plasmapheresis and antiviral drugs. Of different methods of artificial lung ventilation, the best is thought to be transcatheter high-frequency artificial lung ventilation. As shown by the studies of infection resistance, the combined treatment should incorporate such drugs as zovirax and leukomax. Ultrasound disintegration and laser photodestruction with Ho laser were introduced in surgical removal of the papillomas. The combined treatment shortens the treatment duration and prolongs recurrence-free period. PMID- 11011574 TI - [Assessment of the formation of interferon neutralizing antibodies and their influence on the effectiveness of interferon therapy in children with juvenile respiratory papillomatosis]. AB - Formation of neutralizing antibodies (NAB) to alpha-interferon (aINF) was assessed in the reaction of antiviral activity neutralization in 36 children with severe juvenile respiratory papillomatosis (JRP). Surgical removal of papillomas was combined with administration of recombinant aINF preparations (reaferon, intron A, viferon). High and moderate neutralizing activity was determined in 22 (61.1%) patients. 7, 2 and 13 of them received reaferon, intron A and viferon, respectively. NAB were not registered in the rest 14 (38.9%) patients. The frequency of NAB formation was not related to the preparation used, only in the group on reaferon it was insignificantly higher. It was found that the highest response to therapy occurred in children with low NAB titer (8 of 12 children, 66.7%), while children with high NAB titer (3 of 4 children, 75%) did not respond. PMID- 11011576 TI - [Aspects of nasopharyngeal microflora and middle ear function in children]. AB - Exudative otitis media (EOM) was detected in 13 (14.8%) children out of 88 preschool children at audiological investigation. In their nasopharynx, these children carried pneumococci and hemophylic bacilli. The latter were identified 1.7 times more often in children with EOM than in those free of it. Both pathogens were encountered in EOM children 2.6 times more frequently. It is suggested that the above pathogens may support development and maintenance of EOM. PMID- 11011575 TI - [The study of aerobic bacterial background and antibiotic-sensitive isolated strains in the ENT department]. AB - Aerobic microbes in ENT departments were studied. The following strains were isolated: Staphylococcus, Streptococcus, Enterococcus, Acinetobacter, Klebsiella, Pseudomonas, Flavobacter, Citrobacter, Enterobacter, Escherichia, Aspergillus, Candida. 54.5% of the isolated strains were of hospital origin. Comparative evaluation of antibiotic resistance has shown a wide spectrum of resistance of the isolated microorganisms to antimicrobial drugs, among those to the drugs used in the ENT departments. PMID- 11011577 TI - [The evaluation of the palatine tonsil function in patients with chronic tonsillitis]. AB - The authors review the results of the research performed in patients with chronic tonsillitis in 1997-1999. Immunological activity of their palatine tonsils was evaluated according to the original technique which demonstrated relations between external functional activity of the palatine tonsils and age as well as symptoms of chronic tonsillitis. Additional information is presented on interrelations between lymphocytopoietic production of the palatine tonsils and hormonal status, metabolism, ecological and physical factors. The study of lifetime activity of the palatine tonsils is necessary for examination of local immunity of the mucous membranes of the lymphoid pharyngeal ring. PMID- 11011578 TI - [Vertical vestibulo-ocular interaction in unilateral labyrinth lesion (communication II)]. AB - Vertical vestibuloocular reflex (VVOR) suppression was studied in 25 patients with unilateral labyrinthine disorders and 22 healthy individuals. The method of the suppression investigation was based on stimulation of the internal semicircular channels of the labyrinth by active sagittal nodding with glance fixation of the object moving with the head. VVOR suppression by glance reflex in patients with unilateral labyrinthine lesion is characterized by reduced reflex suppression effectiveness, especially on the side of the affected labyrinth, via a significant VVOR phase shift down. PMID- 11011579 TI - [The relationship between the condition of the brain stem compartments of the acoustic analyzer and rheoencephalography in people who participated in the liquidation of the consequences of the Chernobyl' nuclear plant disaster]. AB - Pure-tone and speech threshold and suprathreshold audiometry, short-latent acoustic evoked potentials (SLAEP) and rheoencephalography (REG) were applied in examination of 67 patients with neurosensory hypoacusis (NSH) aged 21 to 45 years. These patients worked as wreckers in the zone contaminated with radionuclides 5 years and later after the Chernobyl accident. Relationships between functional condition of the truncal structures of the acoustic analyser and cerebral hemodynamics were studied. Correlation analysis has shown that there is close correlation between SLAEP and REG, especially in the vertebrobasillar area of cerebral circulation. The closest relations existed between the duration of the latent period of SLAEP wave V, interpeak interval I-V and rheographic index in the vertebrobasillar system. Combination of SLAEP and REG potentialities opens wide prospects for revelation of the key pathogenetic links for disorders of the acoustic system of unconventional origin, improves quality of these disorders diagnosis, promotes individual approach to therapy of such patients. PMID- 11011580 TI - [Otogenic cerebellar abscesses (in the light of 10-year observation materials at the ENT clinic at the Tashkent institute of continuing education of physicians)]. AB - In 1989-1998 ENT specialists of the Tashkent Institute of Postgraduate Medical Education treated 109 patients with otogenic intracranial complications. 13 (11.9%) of them had cerebellar abscess. Clinical symptoms of the abscesses were obscure or absent. Head ache was the leading symptom. Other hypertensive symptoms presented with nausea, (n = 2), vomiting (n = 5), bradycardia (n = 7). Changes on the fundus of the eye were not registered in 6 patients. Defective coordination of movements, scanning speech were observed in 11 and 3 patients, respectively. Large-swinging, mixed horizontal nystagmus was truncal, in 11 patients it was directed to the side of the abscess, in 2 patients--in both directions. Symptoms of the secondary meningitis arose in 12 patients. The patients underwent surgical cleaning of the middle ear and opening cerebellar abscess under drug therapy. One patient died of purulent ventriculitis (lethality 7.6%). It is inferred that cerebellar abscesses often run with mild symptoms. This creates diagnostic difficulties. PMID- 11011581 TI - [Approaches to surgical treatment of snoring and sleep apnea]. AB - Examination and treatment data on 186 patients suffering from snoring and sleep apnea and surgical methods to manage these pathological conditions are reviewed. The stages of the surgical treatment of snoring and sleep apnea are the following: correction of nasal breathing, surgical intervention on the palatine using different tools (laser, scalpel, loop, electric knife). Effectiveness of the treatment methods, complications, postoperative developments are compared. PMID- 11011582 TI - [Prevention and treatment of upper respiratory tract infections with IRS-19 and polydex with phenylephrine]. PMID- 11011583 TI - [The use of controlled hypotonia in endonasal microsurgical interventions on the paranasal sinuses in patients with chronic rhinosinusitis]. AB - The technique of controlled hypotonia was developed for adequate management of anesthesia in endonasal microsurgical interventions on the paranasal sinuses in patients with chronic rhinosinusitis. A total of 92 patients with chronic polypous rhinosinusitis aged 18 to 65 years were operated using the following variants of anesthesia: local, local in combination with neuroleptanalgesia and/or ataralgesia with controlled hypotonia, endotracheal oxide-fluoroxene and endotracheal oxide-oxygen anesthesias with neuroleptanalgesia and/or ataralgesia with controlled hypotonia. This was achieved with clofelin. Controlled hypotonia reduces blood loss in endonasal microsurgery on the paranasal sinuses in patients with chronic polypous rhinosinusitis compared to patients unexposed to controlled hypotonia. PMID- 11011584 TI - [Nasonex effectiveness in allergic rhinitis and chronic polypous rhinosinusitis]. PMID- 11011585 TI - [Long-term presence of the foreign bodies in the maxillary sinuses and nasal cavities]. PMID- 11011586 TI - [Psychogenic aphonia which has developed in a female with unilateral paresis of the recurrent nerve]. PMID- 11011587 TI - [Current aspects of surgical treatment of Meniere's disease]. PMID- 11011588 TI - NPs and the Internet. PMID- 11011589 TI - Easing a mother's worries. PMID- 11011591 TI - Movin' on up. PMID- 11011590 TI - To the test: assessment skills. PMID- 11011592 TI - Renegotiating your salary: words for the wise. PMID- 11011593 TI - Building bone. PMID- 11011594 TI - Uneven ground. HIV in women of color. PMID- 11011595 TI - On their own. School and community management of pediatric asthma. PMID- 11011596 TI - Chronic otitis media in children. Helping families cope. PMID- 11011597 TI - Community-acquired pneumonia. The 'captain of death' can be controlled. PMID- 11011598 TI - Patient information. Preventing adverse drug reactions. PMID- 11011599 TI - Alternative therapies. Tools for the new millennium. PMID- 11011601 TI - STD issues in cyberspace. PMID- 11011600 TI - Legal documentation. A case study in basic concepts. PMID- 11011602 TI - Population growth--is it our concern? PMID- 11011603 TI - Time spent in a mentor's garden. PMID- 11011604 TI - Diagnosing irritable bowel syndrome. PMID- 11011605 TI - Focus on fluids. Examining maternal hydration and amniotic fluid volume. PMID- 11011606 TI - Infant abduction. Lowering the risk. PMID- 11011608 TI - HIV and women. Understand your responsibilities; reduce your risk. PMID- 11011607 TI - Managing second-stage labor. PMID- 11011609 TI - Using games to enhance nursing continuing education. PMID- 11011610 TI - Clinton signs legislation to protect breastfeeding. PMID- 11011611 TI - Uncertified opinion. One nurse's brush with certification. PMID- 11011612 TI - Mental health nurses must try to avoid conflict. PMID- 11011613 TI - Is it safe for nurses to make up i.v. infusions? PMID- 11011614 TI - Case 16: limitations of competence. Nurse who undertook a procedure outside her competence. PMID- 11011615 TI - Peripheral i.v. cannula dressings: advantages and disadvantages. AB - There are many peripheral intravenous cannula dressings being used throughout the country. This article is a review of these dressings and looks at those used in the authors' trust as well as other products on the market. The article gives an overview of the advantages and disadvantages of the different dressings, including costs, that allow medical and nursing staff to make an informed choice about the type of dressing they use. The authors felt, given all the evidence, the traditional method of using gauze, tape or mefix was no longer acceptable practice. PMID- 11011616 TI - Care study: providing effective care in palliative nursing. AB - Currently, nurses are under great pressure to demonstrate effective, evidence based patient care. In this article a single case study will be used to demonstrate that care can be based on recent research within a palliative care setting to produce effective outcomes. Research-based interventions, which were used to provide care for a patient who will be referred to as Maria, will be discussed and evaluated to illustrate the contribution that evidence-based practice made to her nursing care. Although it is a single case study and the findings cannot be generalized, it is suggested that this article can be used to demonstrate the need for accurate interpretation of assessment data, illustrate the use of research to support nursing interventions in pain management, constipation and skin pressure damage, and show that empirical evidence alone may not be a sufficient basis for care and that moral issues must be taken into account. PMID- 11011617 TI - Safe, sound and supportive: forensic mental health services. AB - Certain independent inquiries, the failure to implement the care programme approach, the new mental health strategy, the recent National Service Framework for Mental Health (Department of Health (DoH), 1999), and the Fallon Inquiry into the Personality Disorder Unit at Ashworth Special Hospital have all raised many questions about psychiatric care which remain unanswered. The issue of organizing and developing forensic services can be regarded as a key element if safer services are to be a reality. This article offers a model for forensic mental health services which is derived from policy statements, the published strategy, research, and discussion papers. PMID- 11011618 TI - The development of mental health care pathways: friend or foe? AB - Greater emphasis is being placed on mental health services to be more responsive to clinical variation. Care pathways can standardize clinician response to established diagnostic groups such as schizophrenia. This article discusses care pathways using the findings of a research study carried out in London to explore the development and implementation of a care pathway. Respondents encountered many difficulties such as the principles behind individualized care and generic roles and responsibilities in preformulating their work into a care pathway sequence. This article concludes with areas for further application and research. PMID- 11011619 TI - Evaluating expressed emotion and schizophrenia. AB - This article presents another review from the Network for Psychiatric Nursing Research (NPNR) National Journal Club (see Vol 8(15): 1010-12). The article reviewed was Willets and Leff's (1997) 'Expressed emotion and schizophrenia: the efficacy of a staff training programme'. The collated feedback comments from the regional journal clubs is based on the format proposed by Cormack (1991). The views expressed in this article are those of the journal club members and not necessarily those of the NPNR. PMID- 11011620 TI - Providing correct pressure-relieving devices for optimum outcome. AB - Pressure redistributing (PR) support surfaces are positioned by manufacturers against different levels of patient vulnerability to developing pressure ulcers. However, there is no agreed method through which such linkage between a patient and an appropriate support surface can be made. This lack of clarity leads to new devices being positioned in a manner not representative of their eventual use in clinical practice. Using a data hierarchy approach, three strands of evidence (interface pressure, a pilot national survey and a randomized controlled trial) are presented to illustrate, and support, the repositioning of the Viaclin dynamic mattress overlay (formerly the Pegasus Overture) from use with medium risk patients to use with higher-risk patients. The need for amendment is seen most strikingly in the two clinical studies featured in this article, during which 87% of all patients nursed on the Viaclin dynamic mattress overlay were at high- to very high-risk of developing ulcers. PMID- 11011621 TI - Transcultural competence: the challenge for nurse education. AB - Several recent health policy directives highlight the need to overcome the health disadvantage and discrimination experienced by many members of minority ethnic communities. However, there is ample evidence to indicate that nurses frequently fall short of providing sensitive and appropriate care to minority ethnic patients. If these policy objectives are to be met, it is essential that nurse education provides students with the opportunity to develop transcultural competence. This article examines the current policy context before considering the principles underpinning the development of transcultural competence. It concludes by discussing how nurse education might more effectively prepare nurses to meet the needs of minority ethnic communities. PMID- 11011622 TI - Confidentiality. 11: terminal illness and the duty of confidentiality. PMID- 11011623 TI - Wound irrigation techniques: new Steripod normal saline. AB - Wound cleansing is performed on a regular basis by many healthcare professionals. However, the modern concept of cleansing has not always been a priority in wound management. There are many questions which challenge those responsible for cleaning wounds such as how often a wound should be cleaned and which techniques should be used. This article begins with a brief guide to wound cleansing and then focuses on the new Steripod normal saline produced by SSL International. PMID- 11011624 TI - General nursing does not seem to have progressed. PMID- 11011625 TI - Year 2000. Technology of the future and rediscovering home haemodialysis. PMID- 11011626 TI - Improving adequacy improves haemodialysis outcome. AB - Our goal of maintenance dialysis for the coming millennium is optimal rather than just adequate dialysis. Delivering a large amount of dialysis expressed in terms of urea Kt/V is a necessary but insufficient measure to improve clinical outcome. Cardiovascular morbidity and mortality remain very high in haemodialysis. This is due in great part to the insufficient control of extra cellular volume and blood pressure. Ours, as well as published data, indicate that up to now, only increasing dialysis time either by prolonging the session or increasing its frequency has proven value in overcoming this critical issue. PMID- 11011627 TI - Review of arteriovenous fistula care. AB - The patient with End Stage Renal Failure is largely dependent upon haemodialysis for survival. Vascular access is the vital lifeline. This paper provides a review of the literature and aims to advocate the advantages of arteriovenous fistulae compared with other vascular access. It will discuss the main causes of thrombosis in arteriovenous fistula failure, measurements of early detection of venous stenosis and the care of an arteriovenous fistula to ensure its patency. The responsibility of the dialysis nurse is also considered. PMID- 11011628 TI - Influence of post haemodialysis sampling on Kt/V result. AB - The recommended Kt/V is 1.2. Unfortunately there is no written policy for nurses on the procedure for taking blood urea nitrogen samples post haemodialysis. The aim of this study was to establish the Kt/V variability of haemodialysis patients depending on the method of collection of post-haemodialysis blood urea nitrogen. Twenty-two patients were analysed. A Kt/V was performed every 15 days during a period of 2 months. It was taken five times on each patient: 30 minutes before the end of a haemodialysis session (Kt/V30), at the end of haemodialysis (Kt/V1), after slowing flows (50 ml/min) for 2 minutes (Kt/V2) and after the blood circuit had been returned to the patient at 5 and 15 minutes respectively. (Kt/V5, Kt/V15). The Kt/V results were: Kt/V1 1.23 +/- 0.2 Vs Kt/V2 1.14 +/- 0.19 (p < 0.003); Kt/V5- 1.05 +/- 0.19 (p < 0.002 Vs Kt/V2); Kt/V15 1 +/- 0.16 (p < 0.05 Vs Kt/V5); Kt/V30 1.12 +/- 0.21 (pNS Vs Kt/V2). In conclusion, there was a large variability in the Kt/V depending on the method of collection of the blood urea nitrogen sample post-haemodialysis. PMID- 11011629 TI - One haemodialysis unit's experience of hepatitis B. AB - Haemodialysis patients are at high risk of contracting Hepatitis B Virus (HBV). This blood borne virus has been responsible for many critical incidents within haemodialysis units around the world. This paper seeks to explore one such outbreak and at the same time look more closely at HBV and connected issues. The incident discussed occurred within a haemodialysis unit situated in the Northwest of England. This paper is presented as a case study. When HBV was detected in a previously HBV negative patient a crisis management team moved quickly to contain any possible outbreak and sought to discover the origin of the infection. An outline of the personal and financial cost to the organisation, what lessons have been learnt and why recommendations have been made to immunise the local dialysis population again HBV will be presented. PMID- 11011630 TI - Heparinisation in chronic haemodialysis treatment: bolus injection or continuous homogeneous infusion? AB - After several incidents where patients either bled after dialysis or suffered severe anaphylactic reactions caused by intravenous heparin injections, we decided to administer heparin as a continuous homogeneous intravenous infusion. This had been usual practice in our unit in the past. Following the shocking experience of the death of a patient due to an anaphylactic reaction during his first dialysis, we wanted to warn colleagues for the potential dangers of administering intravenous heparin as a bolus injection. PMID- 11011631 TI - Experiences of patients called to hospital for a renal transplant. AB - When patients develop end stage renal failure (ESRF) the effects can be devastating. Patients perceive transplantation as a solution to their many problems and it is often the treatment of choice. The aim of this study was to examine the experiences of patients with ESRF when called to the hospital for a renal transplant. Semi-structured interviews were taped, transcribed and a thematic data analysis carried out. Results demonstrated that patients felt they were given adequate support on arrival at hospital but they found the wait to determine if they were a suitable match for the kidney difficult. There was evidence that some patients coped with the situation by shutting out negative possibilities. PMID- 11011632 TI - Evaluation of potential transplant recipients and living donors. AB - The transplant waiting list needs to be actively managed by the transplant centre in a systematic way to achieve any degree of efficiency and effectiveness. This paper describes the key role of the transplant nurse in a large centre with over 300 people on the transplant waiting list. It focuses on the way patients are assessed, how the list is updated and the benefit of nurse involvement in the work-up of potential live donors. PMID- 11011633 TI - EDTNA/ERCA dialysis technology Journal Club. Microbiologic quality of dialysis water. PMID- 11011634 TI - Another choice for the blind. AB - Automated Peritoneal Dialysis has proved to be an effective treatment for many patients. This presentation outlines the training issues and management of a 48 year-old registered blind, insulin dependent diabetic, who developed renal failure. PMID- 11011635 TI - Nutritional education of CAPD patients and media influence. AB - The aim of our paper was to investigate the short period of training offered to peritoneal dialysis patients. We focused our attention on the nutritional knowledge of patients who often gather information, and advice from external factors such as their family and the media. We organized group meetings on nutrition for 22 patients and 17 relatives. Questionnaires were completed before and after the education programme. The group meetings improved knowledge on nutrition and highlighted the influence of the media. The important role of the nurse at these meetings has also been recognised. PMID- 11011636 TI - Complications of complicated grief in renal failure. AB - This paper sets out to bring together recurrent themes in the understanding and care of patients with Chronic/End Stage Renal Failure centering on the key issue of loss. A multi-disciplinary conceptual framework is proposed based on three perspectives--Systemic, Existential and Narrative. The integrated framework for understanding the complexity of grief issues for dialysis and pre-dialysis patients focuses on the perception of self and the inter-relationship with others directly involved in their care. PMID- 11011637 TI - Quality of life and exercise in renal disease. AB - Many publications describe the advantages of physical exercise in improving quality of life. The aim of this study was to highlight the need for physical activity when patients commence haemodialysis. A correlation was found between muscular deterioration and the years on haemodialysis and it was accentuated in men. From our study we recommend adequate physical activity at the beginning of, and during the lifetime of haemodialysis to enhance the physical and psychological well being of patients. PMID- 11011638 TI - Hospice nursing--taking charge of our future. PMID- 11011639 TI - Disease management--carpe diem home care! PMID- 11011640 TI - Critical competencies for nurses in the new millennium. PMID- 11011641 TI - Advanced practice nurses in home care are essential. PMID- 11011643 TI - A home healthcare nurse's wish list: version 2000. PMID- 11011642 TI - Behavioral health home care 2000: integration and collaboration. PMID- 11011644 TI - Major home health regulatory changes in 2000. PMID- 11011645 TI - Palliative care ... 2000. PMID- 11011646 TI - Evidence-based practice for home healthcare nurses in the 21st century. PMID- 11011647 TI - Accreditation in the new millennium. PMID- 11011648 TI - The "R" word and the new millennium. PMID- 11011649 TI - Challenges for home healthcare nurses in the 21st century. PMID- 11011650 TI - Establishing a bond with clients of different cultures. AB - In nursing, it is well known that establishing a successful nurse/client relationship depends on the nurse's ability to promote a bond of trust between them (Arnold & Boggs, 1995). A home care nurse working with a client from a different culture will need to be mindful and take the extra steps mentioned in this and other articles. Such steps will help promote this bond of trust and aid the nurse in providing more culturally competent care. However, because every person is unique, these same approaches and structured questions can be asked of all patients. To do so will enable the nurse to have a more complete understanding of each patient's health care beliefs, practices, and decision making strategies. As has been shown through the case studies presented, gaining a more thorough understanding of the patient and his/her family's health care beliefs is critical to achieving cost-effective and clinically positive outcomes. In each of the examples discussed, if these cultural assessments had not been performed, more nursing resources and longer-term service would have been required. PMID- 11011651 TI - Home Talk/Healthy Talk: improving patient's health status with telephone technology. AB - This article describes Home Talk and Healthy Talk, two practical applications of telephone technology developed at the VNA of Cleveland. Home Talk is a disease specific monitoring system with preprogrammed automated questions that immediately score patient responses. If any responses fall out of the safety zone, the system calls the supervisor and a live intervention follows for assessment and appropriate action. Healthy Talk is a series of age-specific automated health screens used by nurses in community centers to engage clients to follow through with self-selected health goals. The use, cost-effectiveness, and impact on outcomes are also discussed. PMID- 11011652 TI - New Joint Commission home care standards and changes for 2000. PMID- 11011653 TI - Are you Y2K ready? PMID- 11011654 TI - Home Care: No Boundaries. PMID- 11011655 TI - "What a long, strange trip it's been". PMID- 11011656 TI - Cognitive behavioural group anger management for out-patients: a retrospective study. AB - This study proposes that anger as the "forgotten emotion" (DiGiuseppe 1994), can cause distress and hardship for the individual who experiences dysfunctional anger in the form of high levels of intensity, duration and rapid onset as well as causing distress to those who observe or are victims of the behavioural responses to anger It is very difficult to calculate the cost of dysfunctional anger to an individuals physical well being and relationships, or to society in general. The author suggests the costs are immense. With this in mind anger management training has gradually emerged in differing forms and from different psychotherapeutic alliances. However, very little outcome data is available to clinicians which may highlight the benefits or deficits of such programmes. To this aim, and using Novaco's (1994) model of anger the author has attempted to address this issue. Twenty three subjects (Ss) attended for screening assessment six group treatment sessions and a three month follow-up session of anger management based on Cognitive Behavioural approaches. The groups were small in number of between 3-6 Ss. All the Ss were from the out-patient, primary care setting, and none of them were compulsory required to attend the sessions. Quantitative data was collected using the 1994 Novaco Anger Scale, (NAS), and scientifically analysed. Qualitative data was collected using a semi-structured post treatment interview, and some of the comments are discussed. The quantitative data showed significant gains across all measures from pre to post treatment. No gains were made during the period spent (3-6 months) on the waiting list. The results and findings are discussed in relation to therapeutic value and service planning. PMID- 11011657 TI - On the importance of congruence between psychiatric staff and patient about the patient's status. AB - This study explores the usefulness of a method for comparing the congruence of the staff's and the patients' perceptions of the patients' well-being. At a psychiatric rehabilitation clinic, a comparison was made between the patients' and the staff's views of the patients' well-being. The staff and the patients completed questionnaires about the patients' well-being on ten occasions during a 5-year period. Through a principal components analysis of the questionnaire scores, two conceptually similar dimensions (health-illness, and contact distance) were identified on each set of scores. The staff and patient ratings on the two dimensions could thus be compared. During the time of the study, a reorganisation of the clinic was made. This made it possible to compare older and newer units. The results indicated that the staff at long-existing units rated the patients' health as lower than the patients did, whereas the staff at newer units rated the patients' health as better than the patients did. The staff at units where the patients lived in their own apartments saw the patients as less contact-seeking than the patients did, and the staff at in-patient units saw the patients as more contact-seeking than the patients did. The two-dimensional PC models made it possible to visually illustrate the developments for three individual patients. The results thus showed that the method is useful for research purposes as well as for clinical evaluations. In the final section, problems and assets of the method are discussed. PMID- 11011658 TI - The milieu and milieu therapy in adolescent mental health nursing. AB - The milieu is a well discussed concept in the mental health nursing literature; so too is milieu therapy. Nevertheless, despite evidently clear understandings of the milieu's significance, and of its therapeutic potential, little published mental health nursing research focuses on these areas of practice. As a result, within the specialty, theoretical conceptualizations of the milieu remain unchallenged and theory development remains in its infancy. Findings from an inquiry into the practice of adolescent mental health nursing demonstrate the: (1) nature, (2) construction, (3) maintenance, (4) function, (5) therapeutic effects, and (6) outcomes of the milieu. Through these findings, present theoretical conceptualizations of the milieu are examined and expanded, and a beginning theory of the adolescent milieu, and of milieu therapy, is presented. PMID- 11011659 TI - Stop, look, and listen: the challenge for children with ADHD. AB - Attention deficit-hyperactivity disorder (ADHD) is a common neuropsychiatric disorder that appears in 4% to 13% of the U. S. population. Contrary to earlier beliefs, ADHD frequently persists into adolescence and adulthood and is associated with poor health choices and risk-taking behavior. Despite a vast amount of scientific literature addressing the disorder, many misconceptions and myths prevail creating confusion for children, their families, and health professionals as they attempt to make treatment decisions. One new theoretical model of ADHD shows promise in its ability to provide a more comprehensive explanation of the disorder; it could prove useful in the development of interventions and research programs to improve health maintenance and decrease health-risk behavior in the ADHD population. PMID- 11011660 TI - Problems related to the diagnosis and treatment of adolescents with leukemia. AB - This descriptive study investigated the problems related to the diagnosis and treatment of adolescents diagnosed with leukemia. Subjects were 8 females and 22 males diagnosed with leukemia for at least one year and who received outpatient treatment. The study found that the information needs of the adolescents related more to the treatment and the prognosis of the disease than to the etiology. All subjects reported that they experienced physical problems relevant to the disease and the treatment, and 50% reported that they experienced social problems. Also, their social problems decreased as the time period after the diagnosis increased and subjects were in the remission stage. Adolescents also have fears and concerns related to the side effects of treatment and prognosis. Especially in the recurrence stage, their fears and concerns about death are increased. Friends and family relationships of the adolescents changed, and their future life plans were disrupted after the disease. PMID- 11011661 TI - Experience of teenagers born with cleft lip and/or palate and interventions of the health nurse. AB - The objective of this study was to describe and understand the experiences of teenagers with cleft lip and/or palate. The human ecology approach of Bronfenbrenner (1979) was chosen as a basis for this study. From this perspective, the living environment of teenagers with cleft lip and/or palate was considered, and as such, qualitative research from an ethnographic standpoint was conducted. The data were collected by semistructured interviews and recorded on magnetic tape. Ten teenagers, between 15 and 17 years of age, were chosen and consulted while undergoing therapeutic treatment and surgery in a cleft lip and/or palate clinic. The results of this study indicate that teenagers born with this congenital malformation experience various physical, functional, or interactional problems throughout their development. To improve this situation, nursing care has been developed to enable the nurse to act within the multidisciplinary team treating cleft lip and/or palate cases, thereby allowing the teenager to develop in an environment that offers a better quality of life. PMID- 11011662 TI - Understanding sibling adaptation to childhood cancer. AB - Clinical research conducted over the past 40 years has described adjustment difficulties healthy siblings face when they have a brother or sister with childhood cancer. Research on healthy siblings of children with pediatric malignancies has advanced over the past few years and has broadened from distinguishing psychosocial problems following the patient's death to identifying stressors during the illness experience. More recent research endeavors have focused on recognizing what behaviors or interventions health care providers understand to be most significant in promoting the patient's, parents', and siblings' coping efforts with childhood cancer and its treatment. In addition, descriptive research is exploring interventions used by pediatric oncology health care providers to render social support to siblings of children with cancer (Murray, 1999). Despite the growing body of literature on sibling adaptation to childhood cancer, an understanding of this experience from a developmental perspective is lacking. This lack of theoretical understanding may contribute to inadequate care of siblings of children with cancer. Understanding the meaning cancer has for well siblings is critical. This article discusses the theoretical framework of child development in relation to understanding sibling adaptation to the childhood cancer experience. PMID- 11011663 TI - Developmental Stifling: an emerging symptom pattern. AB - "Developmental Stifling" is a term recently developed by us to denote a symptom pattern observed in a variety of clinical practice sites. The purpose of our paper is to define developmental stifling and compare this recently identified condition with Munchausen Syndrome by Proxy. A case example of developmental stifling is provided, as well as recommendations for early identification and treatment. PMID- 11011664 TI - Calcium signalling mediated by the 9 acetylcholine receptor in a cochlear cell line from the immortomouse. AB - 1. We have investigated the characteristics of the alpha9 acetylcholine receptor (alpha9AChR) expressed in hair cell precursors in an immortalized cell line UB/OC 2 developed from the organ of Corti of the transgenic H-2Kb-tsA58 mouse (the Immortomouse) using both calcium imaging and whole-cell recording. 2. Ratiometric measurements of fura-2 fluorescence revealed an increase of intracellular calcium concentration in cells when challenged with 10 microM ACh. The calcium increase was seen in 66 % of the cells grown at 39 degrees C in differentiated conditions. A sm aller fraction (34%) of cells grown at 33 degrees C in proliferative con ditions responded. 3. Caffeine (10mM) elevated cell calcium. In the ab sence of caffeine, the majority of imaged cells responded only once to A Ch presentations. Pretreatment with caffeine ingibited all calcium respo nses to ACh. 4. In whole cell tight-seal recordings 10 microM ACh activa ted inward current was dependent on the extracellular calcium concentrat ion with an estimated PCa/PNa of 80 for the alpha9 receptor at physiological calcium levels. 5 . The data indicate that ACh activates a calcium-permeable channel alpha 9AChR in UB/OC-2 cells and that the channel has a significantly higher c alcium permeability than other AChRs. The results indicate that the alp ha9AChR may be able to elevate intracellular calcium levels in hair cell s both directly and via store release. PMID- 11011665 TI - Pathology of femoral head collapse following transtrochanteric rotational osteotomy for osteonecrosis. AB - We investigated the pathology of femoral head collapse following transtrochanteric anterior rotational osteotomy. Six femoral heads were obtained during total hip arthroplasty some 2-12 years after osteotomy. In all cases, the preoperatively necrotic lesions exhibited mostly osteonecrosis with accumulation of bone marrow cell debris and trabecular bone with empty lacunae, although repair tissue such as granulation tissue and appositional bone formation were observed in limited areas in some cases. In the transposed intact articular surface of the femoral head, osteoarthritic changes such as fissure penetration to the subchondral bone and osteophyte formation were commonly observed. In newly created subchondral areas at weight-bearing sites, trabecular thickness and the number of trabecular bones had decreased, with few osteoblasts, osteoclasts, and osteocytes being present, resulting in a coarse lamellar structure of the trabecular bone. These findings suggest that transposed areas in cases of failure consist mostly of low-turnover osteoporotic lesions which could cause collapse of the femoral head. PMID- 11011666 TI - Shockwave application in calcifying tendinitis of the shoulder--prediction of outcome by imaging. AB - This prospective study examined 62 patients (65 shoulders) with chronic courses of calcifying tendinitis of the shoulder before and after low-energy extracorporeal shockwave application (ESWA) in order to identify variables associated with the outcome of this treatment. Before ESWA, radiographs and contrast-enhanced magnetic resonance imaging (MRI) of the affected shoulders were obtained in order to document the size and morphology of the calcifications and the contrast media reactions in areas of interest (deposit, synovia, bursae), respectively. In addition, a clinical evaluation was performed. After ESWA (mean follow-up 18.2 months), clinical evaluations of all 65 shoulders revealed an increase in the Constant score from 44% to 78% (p < 0.0001). While size (p = 0.61) and morphology (p = 0.7) of the deposits before ESWA were not associated with the clinical outcome, negative contrast reactions around the deposits (p) = 0.0001), synovia (p = 0.0049) and bursae (p < 0.01) were associated with improved clinical outcomes. After the total study group was divided into two groups, one with Constant scores > or = 75% (n = 43) and the other with scores < 75% (n = 22), the positive predictive value (ppv), specificity (sp) and sensitivity (se) were determined for the negative reaction around the deposit (ppv: 0.94; sp: 0.95; se: 0.38), synovia (ppv: 0.84; sp: 0.82; se: 0.49) and bursae (ppv: 0.86; sp: 0.86; se: 0.44). In 5 cases (7.7%), surgery of the affected shoulder during the follow-up period was performed. No major side-effects were seen in the study group. In conclusion, our results suggest that in patients with chronic calcifying tendinitis, the absence of contrast enhancement, especially around the deposit, is a strong predictive parameter of a positive clinical outcome of ESWA. PMID- 11011667 TI - Femoral nerve palsy in hip replacement due to pelvic cement extrusion. AB - We report a case in which cement protrusion into the pelvis led to a major complication. During reaming and preparation of the anterosuperior acetabulum, a bony defect resulted which made fixation of an uncemented cup impossible, and a cemented polyethylene cup was used instead. After surgery the patient suffered a complete loss of femoral nerve function. Postoperative X-rays and CTs showed that a huge mass of bone cement protruded into the pelvis in close proximity to the iliac vessels and the femoral nerve. This probably led to nerve damage during the cement's polymerisation process due to the heat. PMID- 11011668 TI - Morphologic and morphometric study of patellar resurfacing with woven carbon filamentous pads. AB - Analysis of retrieved woven carbon filamentous pads, used for resurfacing of the patellar joint surface, disclosed a 4-zonal organizational pattern. Zone 1, facing the articular cavity, was devoid of carbon filaments and consisted of fibrous tissue. Foreign body granulation tissue and fibrous tissue occupied about one-third and approximately 50%-60% of the interfilamentous space in zones 2 and 3, respectively. Carbon filaments formed 2%-9% of zone 2 and 14%-16% of zone 3. An interfacial membrane-like zone 4 separated the carbon filamentous pads from a trabecular bony shell. The bone volume within the latter was approximately 25%. Given that the purpose of articular resurfacing with implants is repopulation of the defect by chondrocytes producing a cartilaginous matrix, the woven carbon filamentous pads did not fulfill this expectation. In an environment of an ongoing foreign body-induced granulomatous reaction, the stem cells permeating the interstices of the woven carbon filamentous pad are apparently incapable of maturing into highly differentiated cells (chondrocytes) synthesizing a highly complex (cartilaginous) matrix. PMID- 11011669 TI - Recurrent intrapelvic cyst complicating metal-on-metal cemented total hip arthroplasty. AB - Spontaneous intrapelvic masses causing vascular compression raise the suspicion of a neoplasm or infection. We present a patient who developed a recurrent intrapelvic cyst 14 years after a McKee-Farrar total hip arthroplasty which presented as acute onset of lower limb swelling, threatening the vascularity of the limb. This cyst recurred three times, and the patient finally needed revision total hip arthroplasty. The most probable cause of the recurrence was the cement and metal wear debris. Such cysts have been described in the literature, but to the best of our knowledge, this is the only report that describes it complicating a metal-on-metal prosthesis. PMID- 11011670 TI - The epidemic of ankle fractures in the elderly--is surgical treatment warranted? AB - Ankle fractures in the elderly are extremely common (up to 184 fractures per 100,000 persons per year, and of these approximately 20%-30% occur in the elderly). The medical literature contains no research that has investigated ankle fractures in the elderly. A prospective, randomised study was conducted of 84 patients with displaced ankle fractures, who were over the age of 65 years and were assigned to operative or conservative treatment after closed reduction. The results of treatment assessed according to the American Orthopedic Foot and Ankle Society (AOFAS) Score showed a mean of 91.37 +/- 8.96 in the non-operated group compared with 75.2 +/- 14.38 (P = 0.001) in the operated group. The costs of treatment were accordingly higher. These results call for consideration of a non operative approach to the treatment of well-reduced ankle fractures in the elderly. Increased efforts should be invested in the prevention of these common fractures. PMID- 11011671 TI - Localization, detection and guided removal of soft tissue in the hands using sonography. AB - Penetrating wounds and lacerations are frequent pathologies treated in the emergency room. The management of hand trauma represents a large part of the work in any surgical practice. Although X-rays are routinely taken, numerous foreign bodies remain undetected, and the wounds are just locally debrided and the lacerations sutured. Unfortunately, as not all foreign bodies are radio-opaque, the radiography results may appear normal, but the patient fails to recover. Patients complaining of persistent wound tenderness were sent for ultrasound investigations, and foreign bodies were detected. Had ultrasonography been carried out initially in the emergency room, the correct diagnosis would have been made, and the sonographic equipment could have helped to guide the physician in his attempt to remove the foreign body. Usually, in response to continued pain, an ultrasound investigation is ordered, and the pathology becomes apparent. A number of examples are briefly described in order to highlight the present inadequacies. It is suggested that hospital administrators consider the need to provide ultrasonographic services as an integral facility of the emergency room. PMID- 11011672 TI - Reduction of blood loss using tranexamic acid in total knee and hip arthroplasties. AB - There have been several attempts to reduce postoperative blood loss in patients undergoing total arthroplasty. Benoni et al. reported the usefulness of tranexamic acid in total knee arthroplasty (TKA). We investigated its effect in TKA and total hip arthroplasty (THA). Blood loss was significantly reduced in patients given tranexamic acid in both the TKA and THA groups, and no severe complications, such as venous or pulmonary thrombosis, were noted in any of the patients who received the agent. Administration of tranexamic acid seems to be useful for reducing postoperative blood loss in TKA and THA. PMID- 11011673 TI - Hyaluronan in synovial fluid of patients with loose total hip prosthesis. Comparison with hyaluronan in patients with hip osteoarthritis and idiopathic osteonecrosis of femoral head. AB - The concentration and molecular weight of hyaluronan (HA) in the synovial fluid of the hip joint were determined in 13 patients (aged 62.8 +/- 9.4 years) who had undergone prior total hip arthroplasty(THA), 23 patients (aged 65.0 +/- 8.2 years) with osteoarthritis of the hip joint (OA), and 13 patients (aged 40.2 +/- 2.7 years) with idiopathic osteonecrosis of the femoral head (ION). A sample of synovial fluid was obtained during revision THA because of loosening of the total hip prosthesis for the THA group, and during the first replacement surgery or osteotomy for the OA and ION groups. The concentration of HA in the synovial fluid was 0.64 +/- 0.42 mg/ml in the THA group, 1.07 +/- 0.28 mg/ml in the OA group, and 1.30 +/- 0.56 mg/ml in the ION group. The concentration of HA in the synovial fluid of the THA patients was significantly lower than that of the OA and ION patients (P = 0.0156 vs OA, P = 0.003 vs ION). The molecular weight of HA was 309 +/- 88.3 x 10(4) Da in the THA group, 377 +/- 201 x 10(4) Da in the OA group, and 240 +/- 148 x 10(4) Da in the ION group; these values do not differ significantly (P = 0.259 vs OA, P = 0.174 vs ION). Among the THA patients, there was no relation between the concentration of HA and the age of the patient, length of time since the first operation, or type of prosthesis fixation; there was also no relation between the molecular weight of HA and each of these factors. These results suggest that a pseudosynovial membrane is regenerated after THA, and that it produces HA of the same molecular weight as that in patients with OA and ION, although in smaller quantities. PMID- 11011674 TI - Posterior cruciate ligament and coupled posterolateral instability of the knee. AB - We wanted to investigate the role of the posterior cruciate ligament (PCL) in the knee's posterolateral stability and the magnitude of the coupled posterolateral instability with the knee examined at 90 degrees of flexion. The coupled posterolateral instability of the knee was studied by selective ligament cutting in cadaver knees set at 90 degrees. The coupled posterolateral displacement after cutting the PCL was 173% of the intact knee. With an intact PCL, the coupled posterolateral displacement after cutting the popliteus tendon and lateral collateral ligament with the knee at 90 degrees of flexion was 299% of the intact knee. When the PCL was cut together with the popliteus tendon and lateral collateral ligament, the coupled posterolateral displacement was 367%. The PCL plays an important role in the posterolateral stability of the knee, and its injury may cause mild (< 5 mm) to moderate (5-10 mm) posterolateral instability. Thus, in a knee with posterolateral instability, injury of the PCL must be considered. With an intact PCL, the posterolateral instability was very recognizable with the knee at 90 degrees of flexion, and injury to the PCL further increased the posterolateral instability and caused posterior translation of the knee. Therefore, examination for posterolateral instability of the knee should also be performed with the knee at 90 degrees of flexion, which is much easier to perform in a clinical setting. A positive posterior translation rather than posterolateral instability at different knee positions differentiates knees with combined PCL and posterolateral instability from knees with isolated posterolateral instability. PMID- 11011675 TI - Isolated palmar dislocation of the fifth carpometacarpal joint diagnosed by stress X-rays. AB - Isolated palmar ulnar dislocation of the fifth carpometacarpal (CMC) joint is a rare injury which often yields only subtle radiographic findings that may be easily overlooked, especially when there is no associated fracture. We reported a case of an isolated palmar dislocation of the fifth CMC joint, diagnosed correctly by means of simple stress X-rays (traction and axial compression stress views). This method proved to be useful and needs no special equipment. PMID- 11011676 TI - Snapping of the proximal interphalangeal joint due to the lateral band. AB - An unusual case is reported of snapping of the proximal interphalangeal joint (PIPJ) of the right little finger in a 14-year-old girl. The lateral band of the radial side slipped over the peak of the condyle of the proximal phalanx at 40 deg of flexion. The lateral band was cut and sutured to the central slip. At follow-up examination 4 months later, there had been no recurrence. PMID- 11011677 TI - Synovial sarcoma with a large hematoma in the inguinal region. AB - We report the case of a patient with synovial sarcoma and a large hematoma of the inguinal region. The patient underwent tumor resection of the lower 2/3 of the acetabulum after preoperative chemotherapy. Twenty-four months after surgery, she is alive without any relapse and can walk without support. PMID- 11011678 TI - Isolated dislocation of the hamate bone. Case report and review of the literature. AB - A case of isolated dislocation of the hamate bone is reported. Mechanism of the injury and treatment are described including a review of the literature. PMID- 11011679 TI - Operative treatment of destructive spondyloarthropathy of the lumbar spine in a patient on chronic dialysis. AB - We present a patient with end-stage renal disease treated by chronic dialysis, in whom progressive destructive spondyloarthropathy of the lumbar spine was successfully managed by fusion with internal fixation, resulting in relief of pain and restoration of activity. PMID- 11011680 TI - Rotationplasty as a salvage procedure in revision of tumor endoprosthesis of the distal femur--a report of two cases. AB - We performed two rotationplasties (type A1 and A2) as salvage procedures in cases of custom-made endoprosthetic replacements of the distal femur and knee joint. The patients suffered from malignant bone and soft-tissue tumors of the knee, respectively. One of them developed chronic osteomyelitis around both stems of the prosthesis, and three local revisions turned out to be ineffective. The other patient, suffering from parossal osteosarcoma of the distal femur, developed a local recurrence following endoprosthetic reconstruction. The functional abilities following the operation of these two patients suggested that rotationplasty was a viable alternative to above-the-knee amputation in failed or severely complicated total arthroplasty of the knee with massive bone loss. PMID- 11011681 TI - Recurrent chondroblastoma of the upper thoracic spine. A case report and review of the literature. AB - A very rare case of recurrent chondroblastoma of the upper thoracic spine with a follow-up of more than 20 years is presented and discussed with a review of the literature. PMID- 11011682 TI - Iris pigment epithelial cell translocation in exudative age-related macular degeneration. A pilot study in patients. AB - BACKGROUND: This prospective, non-controlled pilot study investigates the practicability of IPE translocation and functional outcome in ARMD patients. Removal of submacular choroidal neovascularization (CNV) in age-related macular degeneration (ARMD) is usually associated with RPE damage and poor visual prognosis. Homologous RPE transplants fail to preserve macular function, possibly due to immune rejection. Instead of homologous RPE, we suggest translocating autologous iris pigment epithelium (IPE), building on earlier evidence from animal and in vitro investigations that IPE can substitute RPE functions in the experimental animal. Immunological cell rejection is avoided. METHODS: Four eyes with well-defined and eight eyes with ill-defined subfoveal CNV were submitted to operation and followed up for a minimum of 6 months. IPE cells were harvested from a peripheral iridectomy. A vitrectomy was performed. Submacular membranes were removed, and isolated IPE cells were injected into the subretinal space. Examinations included ETDRS visual acuity, fluorescein angiography, and SLO microperimetry. RESULTS: All patients underwent successful surgical removal of CNV and subretinal IPE injection. Compared to preoperative visual acuity (20/400 20/100) no significant change was observed after 6 months (20/320-16/80). A change of more than two ETDRS chart lines was defined as significant. One eye with preoperative ill-defined CNV developed a recurrence, leading to reduced visual acuity. In all patients, postoperative fluorescence angiography revealed early hyperfluorescence (window defect) in the surgically denuded area. Central fixation was demonstrated in 50% of eyes. CONCLUSIONS: Preliminary data suggests that IPE translocation in submacular surgery for ARMD can preserve but not improve preoperative visual acuity over 6 months. Functional results are promising compared to submacular membrane extraction alone and RPE transplantation. Continued research on improvement of IPE translocation seems justified. PMID- 11011683 TI - Correlation among refractive, keratometric and topographic astigmatism after myopic photorefractive keratectomy. AB - BACKGROUND: Photorefrative keratectomy can be used to flatten the curvature of the anterior cornea and reduce the myopic refraction of the eye. This leads to unphysiological topographical changes of the cornea and may alter the conditions for examinations of corneal surface topography. The purpose of this study was to check for mutual agreement of three different methods of assessment of astigmatism before and after myopic photorefractive keratectomy (PRK). PATIENTS AND METHODS: Forty-seven eyes of 28 patients (age 32.7+/-6.6 years) following PRK using an 193-nm excimer laser were included in this study. 37 eyes were treated for pure myopia (-4.9+/-2.4 D) and 10 eyes for myopic astigmatism (sphere -2.0 to -7.0 D, cylinder -1.0 to -3.0 D). Preoperatively and at 18 months postoperatively, subjective refractometry, keratometry and topography analysis were performed. The axes of topographic and keratometric cylinder were standardized periodically (180 degrees) with respect to the refractive cylinder axis. RESULTS: Pre- and postoperatively, the absolute astigmatism values correlated highly significantly between all three methods (P< or =0.001). The mean refractive cylinder was 0.65+/-0.61 D preoperatively and 0.46+/-0.41 D postoperatively (P=0.2). The mean keratometric astigmatism was 1.14+/-0.64 D before and 0.94+/-0.50 D after PRK treatment (P=0.2). Among the three methods, the mean topographic astigmatism was the highest (P<0.001) preoperatively (1.31+/ 0.56 D) and postoperatively (1.21+/-0.52 D) (P=0.3). In eyes treated for pure myopia, no difference between pre- and postoperative refractive, keratometric and topographic astigmatism was detected (P>0.5). The axes of both topographic and keratometric astigmatism correlated highly significantly with the refractive cylinder axis (R> or =30.9, P<0.0001). CONCLUSION: Up to 2 years after myopic PRK, the difference between refractive and keratometric astigmatism does not differ from the preoperative value, indicating an even corneal surface. The absolute astigmatism values and the cylinder axis correlated well between subjective and objective methods of astigmatism assessment. Thus, objective measurements may be helpful in determining the cylinder component of best spectacle correction after PRK. However, topographic analysis overestimates astigmatism values systematically before and after PRK. PMID- 11011685 TI - Corneal thickness and corneal endothelium in normotensive subjects with unilateral exfoliation syndrome. AB - PURPOSE: To examine the central corneal thickness (CCT) and corneal endothelium in both eyes of patients with unilateral exfoliation syndrome (EXS). To determine the effect of CCT on the measurement of intraocular pressure (IOP). METHODS: In this cross-sectional clinical study, comparisons were made of CCT (Humphrey Ultrasonic Pachometer), corneal endothelial cells (Keeler-Konan contact specular microscope) and IOP (Goldmann applanation tonometer) between the exfoliative (E) and fellow nonexfoliative (NE) eyes in 40 normotensive patients with unilateral EXS. The CCT was used to obtain a corrected value for the IOP. RESULTS: The E eyes had significantly higher values for CCT (0.528+/-0.030 vs 0.523+/-0.032 mm, P<0.01) and IOP (15.7+/-3.6 vs 14.4+/-2.9 mmHg, P<0.001) than the fellow NE eyes. The paired E and NE eyes did not differ in endothelial cell density (2779+/-540 vs 2870+/-386 cells/mm2), in the coefficient of variation of cell size (0.25+/ 0.03 vs 0.26+/-0.03) or in the frequency of hexagonal cells (80.5+/-6.5 vs 82.0+/ 5.0%). After correcting IOP for CCT, the E eyes still had significantly higher IOP than the NE eyes (15.1+/-4.4 vs 14.2+/-3.7 mmHg, P<0.05). CONCLUSION: Normotensive eyes with early EXS did not have quantitative (cell density) or qualitative (variation in cell size, frequency of hexagonal cells) morphological changes in corneal endothelium, but had higher values for IOP and CCT. After correcting IOP for CCT, the E eyes still had significantly higher IOP than the fellow NE eyes. PMID- 11011684 TI - Optic disc vasculitis. AB - BACKGROUND: Our studies had indicated that optic disc vasculitis (ODV) is a distinct clinical entity. We investigated the presentation and clinical characteristics of ODV and determined the efficacy of systemic corticosteroids in its management. METHODS: From 1973 to 1997, we investigated 32 patients (34 eyes) with ODV. The information was obtained by complete medical and ophthalmic history taking and a detailed ophthalmic examination at the initial and follow-up visits. Non-parametric analysis of demographic characteristics and Cox proportional hazard modeling of treatment outcomes was performed. RESULTS: The most common presenting symptom was blurred vision (in 31/34 eyes--91.2%). Visual acuity was 6/12 or better in 82.4% and 6/30 or better in 94.1% of the eyes. All eyes had an enlarged blind spot, vitreous cells and optic disc edema. Fluorescein angiography demonstrated retinal peripapillary phlebitis in 52.6% of the eyes. Systemic hypertension in young patients (P=0.004) and frequency of smoking (P=0.004 for males, P=0.046 for females) were significantly higher than in the general population. Treatment with systemic corticosteroids improved the rate of resolution of optic disc edema and vitreous cells, reducing median resolution times (P=0.042). There was a tendency towards improved central visual fields with treatment (P=0.09). Final visual acuity was 6/12 or better in 25 (92.6%) and 6/18 or better in all of the 27 eyes followed by us. CONCLUSION: Our study indicates that our cases represent a distinct clinical entity of ODV and not just a potpourri of unilateral optic disc edema of different etiologies. It is a self limiting disease, usually with good prognosis, and benefits from treatment with systemic corticosteroids. The presence of retinal peripapillary phlebitis on fluorescein angiography suggests mild vasculitis of the optic nerve head as the primary pathological mechanism in this disorder. PMID- 11011686 TI - Counterrotation of the globe in macular translocation. AB - BACKGROUND: Macular translocation is a new treatment option in age-related maculopathy. The aim of this study was to report experience with the handling of the sensory problems concerning subjective cyclorotation and binocularity that arise with this technique. METHODS: The data of 33 patients who had undergone macular translocation and counterrotation of the globe by muscle surgery in a first operation, followed by silicone oil removal and sometimes revision of the eye muscles in a second operation, were evaluated. The counterrotation consisted of a partial transposition of the rectus muscles and extensive surgery on the anterior margins of the oblique eye muscles. Subjective monocular cyclorotation, motility and binocularity were tested. RESULTS: In 5 patients after the two operations binocular single vision was proved; in one of these cases there was even stereoscopic function. Seventeen patients excluded one eye: 13 excluded the operated eye, 4 the fellow eye. Eleven patients experienced double vision or tilted images under binocular viewing conditions and thus required further surgery or occlusion of one eye. CONCLUSION: The majority of patients were not hindered by subjective cyclorotation. Nevertheless, in a large proportion of patients binocularity could not be reestablished because of still reduced visual acuity or cyclotropia. The data show that the aim of restoring binocularity can be achieved but in practice is often not attained. PMID- 11011687 TI - Measurement of a novel optic disc topographic parameter, "spikiness", in glaucoma. AB - BACKGROUND: Structural changes in the lamina cribrosa have been implicated in the pathogenesis of glaucomatous optic atrophy, but not observed. This paper presents a novel parameter of topographic variability within the optic disc, termed "spikiness", which may reflect glaucoma-related changes in the lamina. METHODS: Four age-matched groups of normal patients (n=12, mean age 64.8 years) and patients with ocular hypertension (n=14, mean age 63.1), primary open-angle glaucoma (n=11, mean age 70) and low-tension glaucoma (n=15, mean age 66.3) were recruited. Images of normal and glaucomatous eyes from the Heidelberg Retina Tomograph were imported into ERDAS image processing software where the spikiness data (30 consecutive mean surface height values across the base of the optic cup in both the vertical and horizontal meridians) were extracted in a format that facilitated further statistical analysis. RESULTS: Significant differences in topographic variability (spikiness) existed in the vertical (F=3.64, P=0.01) but not the horizontal meridian (F=1.25, P=0.3) through the optic disc. Spikiness was inversely related to Humphrey mean deviation (P<0.05), and cup-disc ratio (P<0.004) and was directly related to nerve fibre layer thickness (P<0.005). Of particular interest was the finding that the spikiness measure was the only optic disc parameter to significantly discriminate low tension glaucoma from primary open angle glaucoma. CONCLUSION: A new measure of surface variability (topography) at the floor of the optic cup has been described. The new index of spikiness may represent a measurement of lamina cribrosa fragility which has been implicated, but not previously estimated, in glaucomatous eyes. PMID- 11011688 TI - Long-term results of pneumatic retinopexy. AB - PURPOSE: To evaluate the long-term outcome (>6 years) of retinal reattachment by pneumatic retinopexy. METHODS: Seventy-eight eyes were examined retrospectively after treatment of a retinal detachment with pneumatic retinopexy. The average follow-up period was 6.35 years. The fundus was examined and visual acuity was documented and compared to that of the fellow eye. Any reoperations were evaluated. RESULTS: In 98.7% of cases permanent reattachment of the retina was achieved. In 85.9% the primary intervention had been successful. However, the retina redetached in 19.4% of these during the first 3 months and in a further 4.5% later during the observation period, requiring reoperation. The average number of operations necessary to obtain permanent reattachment of the retina in the 77 eyes was 1.39. One eye was lost due to phthisis bulbi and had to be enucleated. Six years after the first intervention visual acuity of the eyes without reoperation was 0.57 (+/-0.25 SD) with initially detached macula [fellow eye 0.67 (+/-0.27)] and 0.7 (+/-0.27) with attached macula [fellow eye 0.7 (+/ 0.25)]. Visual acuity of the eyes with reoperation was 0.36 (+/-0.3) [fellow eye 0.51 (+/-0.29)] with initially detached macula and 0.64 (+/-0.26) [fellow eye 0.63 (+/-0.35)] with attached macula. CONCLUSION: Six years after initial pneumatic retinopexy the retina was found to be attached in 98.7% of eyes. In 65.4% this result had been achieved by minimally invasive primary surgery alone. The need for reoperation was evident during the first 3 months in 89% of the cases. The final visual outcome was satisfactory overall. PMID- 11011689 TI - Cellular migration into neural retina following implantation of melanin granules in the subretinal space. AB - BACKGROUND: In some retinal diseases and following transplantation of retinal pigment epithelium (RPE), melanin granules are liberated to the subretinal space. Our aim was to investigate the cellular response to implanted extracellular melanin. METHODS: After pars plana vitrectomy, 17 albino rabbits received a suspension of melanin granules in the subretinal space. Postoperative examination included ophthalmoscopy, color fundus photography, histology using monoclonal antibodies identifying RPE cells (AE1/3), macrophages (RAM 11), B-lymphocytes (CD20) and T-lymphocytes (CD45), and electron microscopy. The follow-up time was 2 weeks, 4 weeks and 6 months. RESULTS: On fundus photographs, the layer of melanin showed focal attenuation with lighter areas at 6 months. Melanin granules were phagocytosed by RPE cells and macrophages at 2 weeks, as identified by monoclonal antibodies. In areas where an abundance of melanin was present, multilayers of macrophages were seen associated with considerable photoreceptor damage. Pigment-laden cells invaded the neural retina. The cellular infiltration of the retina was focal, and when it involved the outer nuclear layer the photoreceptor damage was severe. Electron microscopy demonstrated the presence of melanosomes intracellularly in Muller glia. The process of phagocytosis and removal of melanin granules from the subretinal space was slow and not completed at 6 months. CONCLUSION: Our experiments show that implantation of melanin granules in the subretinal space of albino rabbits may induce a considerable phagocytic cellular response featuring the host's RPE, macrophages and glial cells. The migration of pigment-laden cells into the neural retina was associated with focal photoreceptor damage. PMID- 11011690 TI - Morphological changes of retinal pigment epithelial and glial cells at the site of experimental retinal holes. AB - PURPOSE: To investigate morphological changes of retinal pigment epithelial (RPE) cells and glial cells in the immediate and early phases of restoration at the site of experimental retinal holes. METHODS: We made 0.2-disc-diameter retinal holes with a vitreous cutter in albino rabbit eyes. To assess very early changes of RPE cells, the eyes were enucleated at 0,1,3, and 5 h postoperatively and observed by scanning electron microscopy (SEM). At 1,5, and 7 days after surgery, eyes were enucleated and prepared in thin and ultra-thin sections for observation by light and transmission electron microscopy (TEM). RESULTS: SEM showed the surface of the RPE cells lifting a few hours after surgery at the site of the retinal holes. The tissue filling the retinal hole was amorphous and homogeneous, as viewed by light microscopy. TEM revealed that this tissue consisted of glial cell processes containing many organelles. In addition, the cytoplasm of the cells was relatively dark, while the surface of the filling tissue continued smoothly to the internal limiting membrane. These findings suggested that these cell processes were parts of Muller cells. CONCLUSION: RPE cells and glial cells assembling at the site of the retinal hole may play an important role in retinal hole closure in this experimental model. In addition, RPE cells were morphologically changed in an immediate phase of restoration. PMID- 11011691 TI - Inhibition of lens epithelial cell proliferation by substituted PMMA intraocular lenses. AB - BACKGROUND: Migration and proliferation of lens epithelial cells (LECs) are based on interactions between cellular integrins and extracellular matrix proteins (ECMs). We were able to design polymers substituted with COO- and SO3- groups that counteract LEC proliferation through modification of the integrin/ECM interaction. The objective of this study was to compare the cell proliferation inhibiting properties of differently substituted copolymers with those of a homopolymer. METHODS: Polymers were synthesized by radical polymerization and characterized by nuclear resonance spectroscopy. Second-passage rabbit LECs were cultured either on PMMA homopolymers (control) or on substituted PMMA copolymers. Cells were counted on days 2, 7, and 17. Cell vitality was evaluated by trypan blue staining. Experiments were run in quadruplicate. Statistical analysis was performed using the Wilcoxon test and the Mann-Whitney U-test. RESULTS: Compared with control, cells cultured on copolymers grew to significantly lower cell numbers. The inhibitory effect became evident at day 2 after seeding and persisted throughout the experiment. Different ratios of COO-/SO3- substitutions yielded different grades of inhibition. No toxic effect was seen on rabbit LECs. CONCLUSIONS: The tested copolymers inhibit cell proliferation and are nontoxic. Further evaluation could lead to the development of new intraocular lens materials that prevent secondary cataract. PMID- 11011692 TI - Ballistic transfer of minimalistic immunologically defined expression constructs for IL4 and CTLA4 into the corneal epithelium in mice after orthotopic corneal allograft transplantation. AB - BACKGROUND: Experiments were performed to determine whether corneal epithelium transfected with minimalistic immunologically defined expression constructs for the extracellular fragment of CTLA4 and for interleukin-4 (IL-4) or interleukin 10 (IL-10) is able to modulate an allospecific immune response after orthotopic corneal grafting in mice. METHODS: Six groups of BALB/c (H-2d) mice received a C3H (H-2k) corneal graft and dexamethasone eye drops until day 11. Five groups of BALB/c mice had gold particles delivered into the corneal epithelium by Gene Gun on day 10 after transplantation. In four groups, minimalistic immunologically defined gene expression (MIDGE) vectors were delivered into the corneal epithelium by ballistic transfer. The levels of expressed IL-4 and IL-10 were determined by an enzyme-linked immunosorbent assay (ELISA) in shock-frozen homogenized corneas. The expression kinetics of Gene-Gun-transfected corneas were determined by measuring luciferase in lysed whole corneas at different time intervals. RESULTS: Luciferase expression was detectable during the first 5 days following transfection. ELISA was used to determine IL-4 and IL-10 expression in corneal tissue 36 h after transfection. Ballistic IL-4 and CTLA4 gene transfer significantly prolonged corneal graft survival in comparison with the gold treated control group and the IL-10-treated group. CONCLUSION: The beneficial effect of IL-4 and CTLA4, but not IL-10 gene transfer into the corneal epithelium by MIDGE vectors was demonstrated for the first time in corneal transplantation. PMID- 11011693 TI - Ex vivo canine lens capsular sac explants. AB - BACKGROUND: Lens capsular sac explants from human cadaver eyes were used to investigate posterior capsular opacification (PCO). The purpose of this study was to characterize a similar model using canine tissue and to determine whether transferrin (Tf), transforming growth factor beta-2 (TGF-beta2), and insulin-like growth factor-1 (IGF-1) are secreted by lens epithelial cells (LEC) of these ex vivo sacs. METHODS: The lens from canine eyes was removed by extracapsular cataract extraction, the lens sac dissected free, pinned to a petri dish, and cultured in either serum-supplemented or serum-free medium. Morphologic characteristics and growth rate to confluence on the posterior capsule were studied by phase-contrast microscopy. Vimentin, alpha smooth muscle actin, and panTGF-beta expression by LEC were determined by immunohistochemistry. Tf, TGF beta2, and IGF-1 levels were measured by ELISA in the supernatant of sacs cultured in serum-free medium. RESULTS: The mean time to confluence of LEC onto the posterior capsule was 5.4+/-1.1 days (n=22) and 14.7+/-3.7 days (n=14) for sacs in serum-supplemented and serum-free medium, respectively. Following development of confluence, explants displayed opacification and light scatter from cellular proliferation and capsular contraction. Confluent LEC expressed vimentin, alpha smooth muscle actin, and TGF-beta2, and both Tf and TGF-beta2 were secreted into the culture supernatant. CONCLUSION: Canine lens sac explants have characteristics virtually identical to those of human origin, and appear to be a useful alternative tissue source for this model when human cadaver eyes are unavailable. Tf and TGFbeta-2, but not IGF-1, are secreted by LEC in explanted lens sacs and may influence the proliferation and metaplasia of LEC during the development of PCO. PMID- 11011694 TI - Maculo-papillary branch retinal artery occlusions following the Wada test. AB - BACKGROUND: The Wada test induces short-term anesthesia of one hemisphere by injection of sodium amytal into an internal carotid artery. It is an important presurgical diagnostic tool in epileptic patients. PATIENT: A 22-year-old man with idiopathic epilepsy noticed a shadow in the central visual field of his right eye immediately following a Wada test of the right hemisphere. RESULTS: The patient presented with an occlusion of two small branch retinal arteries and corresponding defects in his visual field. Fluorescence angiography revealed small dense hyperfluorescent spots within the occluded retinal vessels. CONCLUSION: Branch retinal artery occlusions are a possible complication of the Wada test, possibly induced by undissolved contrast medium or sodium amytal. PMID- 11011695 TI - Molecular dynamics estimates of ion diffusion in model hydrophobic and KcsA potassium channels. AB - Molecular dynamics simulations are carried out to obtain estimates of diffusion coefficients of biologically important Na+, K+, Ca2+ and Cl- ions in hydrophobic cylindrical channels with varying radii and large reservoirs. Calculations for the cylindrical channels are compared to those for the KcsA potassium channel, for which the protein structure has recently been determined from X-ray diffraction experiments. Our results show that ion diffusion is maintained at reasonably high levels even within narrow channels, and does not support the very small diffusion coefficients used in some continuum models in order to fit experimental data. The present estimates of ion diffusion coefficients are useful in the calculation of channel conductance using the Poisson-Nernst-Planck theory, or Brownian dynamics. PMID- 11011696 TI - Effect of Adriamycin on the boundary lipid structure of cytochrome c oxidase: pico-second time-resolved fluorescence depolarization studies. AB - The fluorescence dynamics of the dye 3,3'-diethyloxadicarbocyanine iodide (DODCI) was used to probe the microenvironment of cytochrome c oxidase (CcO) and cardiolipin. The dye was partitioned between an aqueous and a hydrophobic phase. The 'bound' and 'free' populations of DODCI could be separated by analysis of the time-resolved fluorescence decay of the dye. The anisotropy decay of the DODCI bound to CcO showed a unique 'dip and rise' shape that was analyzed by a combination of rotational correlation times with time-dependent weight factors for each lifetime component. Rotational dynamics studies revealed the existence of a restricted motion of the dye bound at the enzyme surface. Adriamycin, an anticancer, albeit cardiotoxic drug, was previously proposed to affect the surface structure of CcO, most likely by causing a disorder to the surface lipid arrangement. A drastic change in the rotational correlation time of the dye bound to the enzyme surface was observed, which suggested a depletion of cardiolipin layer due to complexation with the drug. The effect of Adriamycin on cardiolipin was drastic, leading to its phase separation. The present study suggests that the effect of Adriamycin on CcO is primarily a segregation of the cardiolipins. PMID- 11011697 TI - Phospholipid monolayer of plant lipid bodies attacked by phospholipase A2 shows 80 nm holes analyzed by atomic force microscopy. AB - In plant storage tissue, lipid bodies are composed of triacylglycerides and surrounded by a phospholipid monolayer which is stabilized by oleosins. At the onset of lipid body mobilization, cells express phospholipase A2, which partially degrades the monolayer and thus provides access for the subsequently acting triacylglyceride degrading enzymes. Analyzing the lipid body surface by atomic force microscopy we show that, at the stage of maximal phospholipase A2 expression, the monolayer contains holes of approximately 80 nm in width and 2.45 +/- 0.46 nm in depth. Non-contact mode imaging was performed with a lateral resolution of approximately 10 nm and a vertical resolution of less than 0.1 nm. The depth of the holes corresponds to the width of the monolayer, while the width of the channels is sufficiently large to provide access to 100 kDa enzymes, such as lipoxygenase and lipases. PMID- 11011699 TI - A new outlook on the energetics of muscle contraction. AB - Analysis of experimental data on two muscles demonstrates that, in contracting striated muscle, the total rate of ATP splitting, nu(t) (number of ATP molecules split per active myosin head per second), comprises of three separate components: nu(p), which is required for the generation of the contractile force P which is equal to the external load; nu(v) which is devoted to the development of the velocity of shortening V; and nu(w), which is responsible for the production of the mechanical power (PV). Nu(p) is proportional to P and nu(v) to V, which means that the sliding distance is independent of P. The mechanical power was found to be equal to the free energy change associated with the hydrolysis of nu(w), which means that the thermodynamic efficiency of the power-producing component is practically 100%. It is concluded that ATP hydrolysis is actually three different reactions. The analysis leads to Hill's force-velocity relationship. Its empirical constants a and b are expressed by thermodynamic and molecular parameters. The constant a was found to be inversely proportional to the sliding distance. The same considerations and conclusions should apply also to other muscles and to the movement under load of microtubules interacting with, e.g. kinesin. PMID- 11011698 TI - DNA, RNA and hybrid RNA-DNA oligomers of identical sequence: structural and dynamic differences. AB - A 27-mer sequence was synthesised as DNA duplex (DD), RNA duplex (RR), and RNA DNA (RD) hybrid in order to characterise their structural and dynamic features. The hydrodynamic radius (Rh) and the rise (b) values of the three samples were consistent with the conformations predicted by CD analysis. The value of the torsional constant (alpha) of the samples containing RNA was approximately twice that of the DD sample and followed the order: DD < RD < RR. The same order was observed in the thermodynamic stability and in the reduction of the electrophoretic mobility. gamma-Ray footprinting analysis was carried out to resolve the individual strand conformation in the hybrid. The RNA strand preserved its conformation, while the DNA strand showed local deformations mainly at TA and TG steps. PMID- 11011700 TI - Analysis of potentiometric titrations of heterogeneous natural polyelectrolytes in terms of counterion condensation theory: application to humic acid. AB - A model, developed within the framework of the counterion condensation theory of linear polyelectrolytes, is presented in this paper to describe the acid-base properties of linear polyelectrolytes, consisting of several types of functional ionizable groups. This formalism has been successfully applied to Fluka humic acid under salt-free conditions, as well as in the presence of supporting simple 1:1 salt (KNO3) at three different concentrations. As part of this approach, the charge density of the humic acid is obtained from the activity coefficient measurements of potassium counterions at different humic acid concentrations at a constant degree of dissociation of the polyelectrolyte. The humic acid average charge density was found to be 0.80 +/- 0.05. Using the present model, we are able to satisfactorily describe the experimental data obtained from acid-base potentiometric titrations. Four main functional groups making up the polymer are determined through their fractional abundances (Xi) and intrinsic pK (pK0i) values. The fractional abundances remained constant and independent of the ionic strength, indicating that the humic acid constitution does not depend on the concentration of excess salts. The pK0i values show a small change with ionic strength, which can be explained by the polyelectrolytic behavior of the solution. PMID- 11011701 TI - Effect of temperature on viscosity properties of some alpha-amino acids in aqueous urea solutions. AB - Viscosities for solutions of glycine, DL-alpha-alanine, DL-alpha-amino-n-butyric acid, DL-valine, DL-leucine and L-serine in 5 mol kg(-1) aqueous urea have been determined at 278.15, 288.15, 298.15 and 308.15 K. The viscosity B-coefficients for the amino acids in the aqueous urea solution have been calculated at different temperatures. The effect of temperature on the B-coefficients is discussed on the basis of the Feakins equation. The contribution of solute to the activation parameters (delta mu0*2, deltaH0*2, deltaS0*2) for viscous flow of the solution have been calculated, together with the Gibbs energy, enthalpy and entropy of transfer for the amino acids from the ground-state solvent to the hypothetical viscous transition state solvent. The contributions of the charged end group (NH3+, COO-) and CH2 groups of the amino acids to B-coefficient and delta mu0*2 have been also estimated using the linear correlations between B coefficient or delta mu0*2 and the number of carbon atoms in the alkyl chains of the amino acids. All the activation parameters are discussed in terms of the solute-solvent interactions in the ground and transition states. PMID- 11011702 TI - Relating helix tilt in a bilayer to lipid disorder: a mean-field theory. AB - We present a mean-field theory relating the helix tilt angle in a bilayer to lipid disorder. The theory provides a method to compare the rotational barriers for different helices in lipid bilayers. The results suggest that the helix tilt angle is strongly affected by both the hydrophobicity of the helix and the average lipid disorder. This leads us to point out future experiments that could shed light on lipid-protein interactions. PMID- 11011703 TI - A thermodynamical study of the clockwork hypothesis proposed by E. Schrodinger. AB - In the present study, the 'clockwork' hypothesis proposed by Schrodinger was examined from the viewpoint of thermodynamics. Firstly, noticing a unidirectional transfer of entropy in a heat engine, the logic was briefly explained about a close relation between this entropy transfer and an irreversible cycle performed by a working body. Next, paying attention to two fundamental differences between a heat engine and a biological system, we considered an isolated system Asigma consisting of three one-component systems (Ai, A, Ao) and noted a case that the same molecules as the component ones flowed quasistatically into Ai from the outside. Then, the unidirectional flows of the molecules, energy and entropy, which were induced by the above inflow in Asigma, were formulated on the basis of the equilibrium thermodynamics for an open system. Furthermore, it was clarified that the fundamental equation for these flows is the Schrodinger inequality and that the necessary-sufficient condition for this inequality is the existence of an irreversible cycle performed by A. Here A corresponds to a working body in a heat engine. It was, thus, concluded that the 'clockwork' hypothesis by Schrodinger is considered to be reasonable for a biological system composed of various irreversible subsystems. PMID- 11011704 TI - Transpupillary thermotherapy for age-related macular degeneration: long-pulse photocoagulation, apoptosis, and heat shock proteins. AB - OBJECTIVE: To provide a biophysical foundation for using transpupillary thermotherapy (TTT) to manage choroidal neovascularization in age-related macular degeneration (ARMD). METHODS: Retinal temperature rise in laser therapy is proportional to retinal irradiance (laser power/area) for a particular spot size, exposure duration, and wavelength. TTT is a low irradiance, large spot size, prolonged exposure (long-pulse), infrared laser photocoagulation protocol. Results from an experimentally confirmed, finite element model of retinal light absorption and heat conduction are used to analyze laser parameter selection and its consequences. Results from apoptosis, heat shock protein and hyperthermia research are used to examine how chorioretinal damage from clinical procedures might be reduced. RESULTS: Chorioretinal thermal equilibration occurs during long pulse TTT photocoagulation. Retinal temperature increases are similar in the RPE where laser radiation absorption is significant and in the adjacent neural retina where there is negligible radiation absorption. For parameters used to treat occult choroidal neovascularization in lightly-pigmented fundi (800-mW, 810-nm, 3 mm retinal spot diameter, 60-sec exposure duration), the maximum chorioretinal temperature elevation is calculated to be roughly 10 degrees C, significantly lower than the 20 degrees C temperature elevations measured in threshold, conventional short-pulse retinal photocoagulation. CONCLUSIONS: To achieve a preselected temperature rise, TTT laser power must be increased or decreased in proportion to the diameter rather than the area of the laser spot. Clinical power settings should be adjusted for fundus pigmentation and media clarity because both of these factors affect absorbed retinal irradiance and thus retinal temperature rise. Noninvasive thermal dosimetry currently is unavailable for clinical retinal photocoagulation, but potential thermometric techniques include MRI, liposomal-encapsulated dyes, multispectral imaging or reflectometry, and subretinal or episcleral thermometry. TTT may be useful not only as independent therapy, but also as an adjunct to PDT, antiangiogenic drugs and ionizing radiation therapy in the management of neovascular ARMD. Low temperature, long pulse photocoagulation is a potential strategy for decreasing neural retinal damage in subsequent TTT or short-pulse photocoagulation and perhaps even for treating glaucoma or retinal degenerations. PMID- 11011705 TI - Scleral buckling for stage 4 retinopathy of prematurity. AB - BACKGROUND AND OBJECTIVE: Both vitrectomy and scleral buckling have been used to treat patients with stage 4 retinopathy of prematurity (ROP). The standard procedure of scleral buckling for treatment of stage 4 ROP is encircling scleral buckling. The effectiveness of segmental scleral buckling is still unclear. The purpose of this study was to analyze the surgical results of patients with stage 4 retinopathy of prematurity treated with either encircling or segmental scleral buckling. PATIENTS AND METHODS: We retrospectively analyzed the anatomic outcomes of 23 eyes (18 infants) with stage 4 A or B ROP treated with scleral buckling. Segmental buckling was used to treat 15 eyes with detachment limited to the temporal side of the retina, while 9 eyes with detachment involving both the nasal and temporal sides were treated with encircling scleral buckling. RESULTS: Macular reattachment during a mean follow-up period of 34 months (range: 2 weeks to 9 years) was accomplished in 11 (79%) of the 14 eyes treated with segmental scleral buckling and 4 (44%) of the 9 eyes that received encircling scleral buckling. There were no intraoperative complications recorded. None of the eyes that achieved macular reattachment developed recurrent macular detachment during the follow-up period. CONCLUSIONS: Scleral buckling appears to be effective for treatment of stage 4 ROP. In patients with temporal side retinal detachment only, segmental scleral buckling provides adequate treatment for stage 4 ROP. PMID- 11011706 TI - Visual field results after vitreous surgery in a case series of patients with open-angle glaucoma. AB - BACKGROUND AND OBJECTIVE: Visual field loss is a complication of vitrectomy and a concern for patients with glaucoma. Our objective was to determine whether vitrectomy is associated with new field defects in patients with glaucoma. PATIENTS AND METHODS: This retrospective case series involved 7 eyes of 7 patients, who had open-angle glaucoma and underwent vitrectomy for macular hole (2) or epiretinal membrane (5). RESULTS: Acuity improved by at least two lines in 5 eyes. Visual fields showed a change in mean deviation following vitrectomy (mean difference -1.50 dB, P=0.0006, paired Student's t-test). One eye had confirmed progression of a pre-existing visual field defect, but no new defects were apparent in the other eyes. Transient intraocular pressure rises requiring therapy occurred in 3 eyes. One eye developed macular edema, but no other operative complications occurred. CONCLUSIONS: The results of our small case series suggest that vitrectomy may be an acceptable intervention in eyes with glaucoma and co-existing macular problems. PMID- 11011707 TI - Pars plana vitrectomy for macular holes combined with cataract extraction and lens implantation. AB - BACKGROUND AND OBJECTIVE: To evaluate the safety and efficacy of cataract extraction and lens implantation at the time of pars plana vitrectomy (PPV) for macular hole (MH) repair. MATERIALS AND METHODS: A retrospective study was conducted of 21 eyes that had PPV for MH combined with cataract extraction and lens implantation. Visual acuities (VA) were followed and complications noted. RESULTS: Of 21 eyes, successful anatomic closure was achieved in 16 (76.2%), and 13 (62.0%) improved in VA by at least 2 Snellen lines. Few complications occurred. CONCLUSION: Procedures that combine PPV for MH with cataract extraction and lens implantation decrease the need for future ophthalmic surgery, allow for clearer intraoperative visualization, and facilitate visual rehabilitation. This combined procedure is a relatively safe and effective alternative to separate pars plana and cataract surgeries. PMID- 11011708 TI - One-year follow-up of laser trabeculoplasty using Q-switched frequency-doubled Nd:YAG laser of 523 nm wavelength. AB - BACKGROUND AND OBJECTIVE: To investigate clinical efficacy and safety of a new laser treatment of open angle glaucoma that targets pigmented cells in the trabecular meshwork for disruption. MATERIALS AND METHODS: The 16 eyes of chronic open angle glaucoma patients were treated in the method similar to argon laser trabeculoplasty using Q-switched, frequency-doubled Nd:YAG laser (523 nm). The treatment results were followed and analyzed for a one-year period. RESULTS: Mean intraocular pressure reduction of 3.81 mm Hg (15%) at 3 months, 3.94 mm Hg (16.1%) at 6 months, and 4.93 mm Hg (20.2%) at 12 months from the baseline were obtained. Slightly greater reductions were seen in the responder group that consisted of eyes showing 3 mm Hg or greater reduction. Adverse reactions were minimal and not significant. CONCLUSION: This new laser trabeculoplasty has shown reasonable efficacy and safety on a limited number of eyes over a one year duration. PMID- 11011709 TI - Prospective randomized comparison of simultaneous and sequential bilateral photorefractive keratectomy for the correction of myopia. AB - BACKGROUND AND OBJECTIVE: To evaluate the efficacy and safety of simultaneous bilateral photorefractive keratectomy (PRK). PATIENTS AND METHODS: There were 52 patients (104 eyes) with myopia less than -6 diopters entered in this prospective trial and randomized to simultaneous or sequential bilateral PRK. Main outcome measures were uncorrected visual acuity (UCVA), percentage of eyes within +/-0.5 D of attempted outcome, present pain intensity (PPI) and total pain rating index (PRI(R)T) scores resulting from McGill the Pain Questionnaire, subjective symptoms, haze, and patient satisfaction. Follow-up extended to 14 months. RESULTS: There was no significant difference in UCVA (P=0.996) and percentage of eyes within +/-0.5 D of attempted correction (P=0.932). Present pain intensity and PRI(R)T scores were significantly higher in the sequential group (P=0.037; P=0.034). The percentage of satisfied patients after bilateral PRK was significantly higher in the simultaneous group (P=0.036). CONCLUSIONS: Simultaneous bilateral PRK was safe and effective, without any significant difference from sequential treatment. PMID- 11011710 TI - Corneal endothelial damage after neodymium:YAG laser iridotomy. AB - BACKGROUND AND OBJECTIVE: This study examined the long-term effect of neodynium:YAG (Nd:YAG) laser iridotomy on the corneal endothelium. PATIENTS AND METHODS: A prospective study was designed. Patients with narrow and occludable angles or fellow eye of acute angle closure glaucoma attack were treated with Nd:YAG laser iridotomy. For one year, 31 eyes of 21 patients underwent complete follow-up. Corneal endothelial specular microscopy was performed before and after laser iridotomy at 1, 3, 6, and 12 months. RESULTS: The decrease of endothelial cell density after YAG laser iridotomy was statistically significant at 1 month (P = 0.036), 6 months (P = 0.004), and 12 months (P = 0.000), respectively. The decrease was not statistically significant at 3 months (P = 0.467). Linear regression analysis indicated no statistical correlation between the percentage change in endothelial cell density and the total energy used during the treatment (1 month: P=0.08, 3 months: P= 0.3, 6 months: P=0.9, 12 months: P=0.2). CONCLUSION: This study demonstrated significant endothelial cell loss in the 1 year follow-up. The result suggested that Nd:YAG laser iridotomy may pose a long term hazard to the corneal endothelium. PMID- 11011711 TI - Exophiala werneckii endophthalmitis following cartaract surgery in an immunocompetent individual. AB - A case of infectious endophthalmitis caused by the saprophyte Exophiala werneckii is reported. This has not been recognized as a pathogen for ocular infections previously. The infection followed uncomplicated cataract surgery involving phacoemulsification and IOL implant. Clinical presentation was that of an indolent endophthalmitis with relatively acute onset. Pars plana vitrectomy, fungal stains, and culture established the diagnosis. Initial management consisted of empirical intravitreal injection of vancomycin, ceftazidime, and amphotericin B. Treatment was supplemented with a 3-week course of systemic fluconazole and topical therapy with natamycin, atropine, ciprofloxacin, and diclofenac. The visual acuity returned to 20/20-2 with no recurrence of infection. The source of the infection could not be determined. Fungal endophthalmitis has to be considered as a rare, though important, complication following ophthalmic surgery. Specific fungal stains and cultures are helpful for establishing the diagnosis early in the course of disease. E werneckii should be considered in the differential diagnosis of fungal endophthalmitis. PMID- 11011712 TI - Combined hamartoma of the retina and retinal pigment epithelium with full thickness retinal hole and without retinoschisis. AB - Combined hamartoma of the retina and retinal pigment epithelium has been described as a rare, benign tumor formed by an overgrowth of several constituents of the retina such as the retinal pigment epithelial cells, vascular elements, and glial components.1 Despite its varied clinical appearance it is important to recognize this tumor as it is known to simulate intraocular malignancies like retinoblastoma and choroidal melanoma. PMID- 11011713 TI - Pre-Descemet's hematoma in a patient of congenital glaucoma with Descemet's detachment: a unique form of hemorrhage. AB - A unique form of anterior segment hemorrhage called Pre-Descemet's hematoma is a previously unreported type of hemorrhage in which blood collects in the narrow space between the corneal stroma and detached Descemet's membrane, which in our case was because of congenital glaucoma associated with Haab's straie. PMID- 11011714 TI - Phacoemulsification and opaque intraocular lens implantation for the treatment of intractable diplopia. AB - Intractable diplopia secondary to paralytic strabismus may be treated with the occlusion of the affected eye. Established treatments include occlusion with a patch, opaque spectacles or contact lenses, and performing upper lid lowering procedures or tarsorrhaphies. Two cases are presented in which intractable diplopia was resolved by performing clear lens phacoemulsification and the insertion of an opaque intraocular lens. Both patients reported a resolution of their diplopia having been dissatisfied with other forms of treatment. Although opaque intraocular lens insertion is not recommended as the initial treatment of choice in such cases, it offers a valuable treatment option for those patients in whom other forms of therapy have failed. PMID- 11011715 TI - CO2-laser in the removal of a plexiform neurofibroma from the eyelid. AB - We present a child with neurofibromatosis type I (NF-I) who underwent excision of a large upper lid neurofibroma using a CO2-laser. Plexiform neurofibromas are notorious for their bleeding tendency and the inability to achieve complete surgical excision. The CO2-laser is an adjunct to achieving better hemostasis and delineation of the tissue in the absence of natural tissue planes caused by the tumor's diffuse mode of growth. Using the CO2-laser achieves better cosmesis, while reducing operation time and limiting complications. PMID- 11011716 TI - Epithelial folds following slippage of LASIK flap. AB - We present a case of a slipped corneal flap after laser-assisted in-situ keratomileusis, LASIK. A 22-year-old, female with myopia and a refractive error of -5.50 D sph OU had an uneventful LASIK performed on her left eye with a Chiron Automated Corneal Shaper (ACS) and a VISX-Star Excimer laser using a nasal hinged flap. On the first postoperative day, slit lamp examination revealed 4.0 mm slippage of the corneal flap. The flap was rolled and folded on itself within the inferior cul-de-sac. With the aid of the operating microscope, the flap was repositioned on the stromal bed. Immediately following the repositioning, many folds were still visible despite centering the flap on the stromal bed and performing stretching maneuvers. Detailed examination showed the folds to be of edematous epithelial origin. The underlying stromal bed was flat. Within days following repositioning, the folds had disappeared and the flap assumed the typical postoperative LASIK appearance. The final uncorrected visual acuity returned to 20/20. PMID- 11011717 TI - Repairing the hole in hydroxyapatite orbital implants following peg removal. AB - Pegging hydroxyapatite implants may lead to improved prosthetic eye movement but may also be associated with complications that may occasionally require peg removal. Removing the peg and leaving the peg hole to spontaneously granulate in, is one technique to deal with the remaining implant hole. The author describes a simple technique to fill the peg hole that provides a smooth implant-conjunctival prosthesis interface and may potentially allow repegging. PMID- 11011718 TI - Bilateral retinoschisis, retinal neovascularization, and severe myopia in a young female. PMID- 11011719 TI - Performance of capillary columns for high-temperature gas chromatography AB - The developments in stationary-phase synthesis and capillary column technology have opened new perspectives in the analysis of high-molecular-weight compounds (600 daltons) and thermolabile organic compounds by high-temperature-high resolution gas chromatography. This branch of high-resolution gas chromatography deals with analysis performed up to 390 degrees C oven temperature (with some applications going up to 420 degrees C and even a few applications to 450 degrees C maximum). The technique has been applied to many different fields of science (e.g., organic geochemistry, environmental chemistry, archeology, and natural product research). Apolar and medium-polar gum phases can now be operated at temperatures from 400 to 480 degrees C, but these higher temperatures are seldom used because of the thermostability of the material used to make the capillary tubing. This paper shows the performance of nine commercial high-temperature columns when used in routine applications. PMID- 11011720 TI - Measurement of gas-liquid partition coefficient and headspace concentration profiles of perfume materials by solid-phase microextraction and capillary gas chromatography-mass spectrometry AB - An empirical model describing the relationship between the partition coefficients (K) of perfume materials in the solid-phase microextraction (SPME) fiber stationary phase and the Linearly Temperature Programmed Retention Index (LTPRI) is obtained. This is established using a mixture of eleven selected fragrance materials spiked in mineral oil at different concentration levels to simulate liquid laundry detergent matrices. Headspace concentrations of the materials are measured using both static headspace and SPME-gas chromatography analysis. The empirical model is tested by measuring the K values for fourteen perfume materials experimentally. Three of the calculated K values are within 2-19% of the measured K value, and the other eleven calculated K values are within 22-59%. This range of deviation is understandable because a diverse mixture was used to cover most chemical functionalities in order to make the model generally applicable. Better prediction accuracy is expected when a model is established using a specific category of compounds, such as hydrocarbons or aromatics. The use of this method to estimate distribution constants of fragrance materials in liquid matrices is demonstrated. The headspace SPME using the established relationship between the gas-liquid partition coefficient and the LTPRI is applied to measure the headspace concentration of fragrances. It is demonstrated that this approach can be used to monitor the headspace perfume profiles over consumer laundry and cleaning products. This method can provide high sample throughput, reproducibility, simplicity, and accuracy for many applications for screening major fragrance materials over consumer products. The approach demonstrated here can be used to translate headspace SPME results into true static headspace concentration profiles. This translation is critical for obtaining the gas-phase composition by correcting for the inherent differential partitioning of analytes into the fiber stationary phase. PMID- 11011721 TI - High-performance liquid chromatographic determination of glycogen in sea urchin gonads with refractive index detection. AB - A high-performance liquid chromatographic method using refractive index detection for the determination of glycogen in sea urchin (Paracentrotus lividus) gonads is developed. After alkaline digestion with sodium carbonate, samples are adjusted to pH 4.6 with citric acid and incubated with amyloglucosidase to hydrolyze the glycogen. The resulting glucose is determined using a Spherisorb NH2 column as the stationary phase and an acetonitrile-water mixture (80:20, v/v) as the mobile phase. The relative standard deviation (%) was 3.57, the limit of detection was 40.1 microg/mL, and the recovery percentage was 97.2%. PMID- 11011722 TI - Computer simulation for the simultaneous optimization of any two variables and any chromatographic procedure. AB - Computer software that allows the simulation of any chromatographic separation as a function of simultaneous changes in any one or two variables that can affect sample separation order (selectivity) is described. For one example, an application is described for the simultaneous variation of the mobile phase pH and gradient time in reversed-phase liquid chromatography. The accuracy of such predictions is examined for a sample mixture of 17 substituted benzoic acids and anilines, and requirements for an acceptable predictive accuracy are summarized. In a second example, the separation of three peptides by capillary electrophoresis is optimized. PMID- 11011723 TI - Quantitation of a metal deactivator additive by derivatization and gas chromatography--mass spectrometry AB - The quantitative analysis of phenolic and amine-containing petroleum additives can be challenging. One such compound-N,N'-disalicylidene-1,2-propanediamine, a common metal deactivator additive (MDA)--is thought to inhibit fuel oxidation catalyzed by metals both in the fuel and on surfaces. The ability to measure the concentration of MDA in storage stability tests, thermal-stressing studies, and field samples is important. Quantitating low concentrations of MDA can be difficult because of surface adsorptivity due to the phenol and amine functional groups. This paper describes the shortcomings of direct-injection gas chromatography-mass spectrometry to quantitate MDA as well as a solution to the analytical problem using the common silylation agent BSA to derivatize the MDA. Results demonstrate that the silylation technique is suitable for the determination of MDA concentrations in aviation fuel samples and suggests that the MDA may be readily determined in other petroleum products with a lower detection limit for MDA of 0.5 mg/L. Measurements conducted in heated batch reactors indicate that MDA concentration is reduced as hydrocarbon fuels are stressed. In addition, only free or available MDA is measured by this technique, not MDA that is complexed with metals. PMID- 11011724 TI - Extraction of atrazine and its metabolites using supercritical fluids and enhanced-fluidity liquids AB - Supercritical fluid and enhanced-fluidity liquid extractions are performed on spiked sediment samples containing atrazine (ATRA) and five of its metabolites including desisopropyldesethylatrazine, desethylhydroxyatrazine (DEHA), desisopropylatrazine, desethylatrazine, and hydroxyatrazine (HA). The hydroxylated metabolites are of particular interest because of their increased water solubility and the fact that their high polarity makes them difficult to analyze. Soxhlet extractions using methanol are conducted for the purpose of comparison. Results of the extractions show that the hydroxy-containing metabolites of ATRA are not effectively extracted with supercritical CO2 alone. The solvating or desorbing power of carbon dioxide appears too low to extract HA and DEHA. The extraction recoveries of the hydroxylated metabolites increase when enhanced-fluidity liquid mixtures of methanol/CO2 are used, and these rates increase with the methanol concentration. Enhanced-fluidity ternary liquid mixtures of H2O/methanol/CO2 yield the best recoveries for these compounds. ATRA recoveries are equally effective when using supercritical CO2 or enhanced fluidity mixtures. The other nonhydroxy-containing metabolites require the increased solvent strength of either large percentages of methanol in CO2 or ternary mixtures of H2O, methanol, and CO2 for high recoveries. Recoveries with enhanced-fluidity liquid ternary mixtures are better than the recoveries from Soxhlet for all the compounds in the study. PMID- 11011726 TI - How can I clean my column having an alkyl amine IP reagent? PMID- 11011725 TI - Chromatographic properties of the ion-exclusion column IonPac ICE-AS6 and its application in environmental analysis, Part II: application in environmental analysis AB - Biological processes, geochemical reactions, anthropogenic emissions, and transformation reactions of xenobiotics are responsible for the widespread occurrence of aliphatic carboxylic acids in the environment. To study the performance of the ion-exclusion chromatography column IonPac ICE-AS6 in the analysis of environmental and environmental-technical samples, organic acids are investigated in composting seepage, silage effluents, aqueous extracts of sewage sludge, molasses hydrolysate, and alkaline cellulose hydrolysates. With respect to the diverse sample matrix and composition, different chromatographic conditions are applied. It is possible to determine various volatile fatty acids, dicarboxylic acids, (poly)hydroxy acids, and keto acids as main and trace components in samples with very high and low dissolved organic carbon content. Low baseline noise allows the determination of malic and succinic acid in the concentration range of approximately 1 microM/L in the presence of higher concentrations of fully ionized compounds. The applicability of the column in environmental analysis may be limited by the poor retardation of strong organic acids, insufficient separation of some relevant substance combinations (i.e., citric and isocitric acid), and very strong hydrophobic interactions with straight-chain monocarboxylic acids containing four or more carbon atoms. PMID- 11011727 TI - An occupancy factor of unity should always be used for waiting rooms and other highly-occupied public areas. PMID- 11011728 TI - X-ray videodensitometric methods for blood flow and velocity measurement: a critical review of literature. AB - Blood flow rate and velocity are important parameters for the study of vascular systems, and for the diagnosis, monitoring and evaluation of treatment of cerebro and cardiovascular disease. For rapid imaging of cerebral and cardiac blood vessels, digital x-ray subtraction angiography has numerous advantages over other modalities. Roentgen-videodensitometric techniques measure blood flow and velocity from changes of contrast material density in x-ray angiograms. Many roentgen-videodensitometric flow measurement methods can also be applied to CT, MR and rotational angiography images. Hence, roentgen-videodensitometric blood flow and velocity measurement from digital x-ray angiograms represents an important research topic. This work contains a critical review and bibliography surveying current and old developments in the field. We present an extensive survey of English-language publications on the subject and a classification of published algorithms. We also present descriptions and critical reviews of these algorithms. The algorithms are reviewed with requirements imposed by neuro- and cardiovascular clinical environments in mind. PMID- 11011729 TI - Localization of cerebral arterovenous malformations using digital angiography. AB - Since 1989 we performed stereotactic radiotherapy treatments of cerebral arterovenous malformations (AVM), estimating three-dimensional (3-D) localization and shape of target volumes by the Leksell stereotactic helmet on two orthogonal radiographic projections. Due to the limitations of this method, we developed a new technique for the localization of the target volume using digital subtraction angiography (DSA) and digital image processing. To achieve this result we first developed a method to correct nonlinear distortion of DSA images using spatial relocation of image pixels based on a calibration grid. We then developed an algorithm for localization of the target volume using two independent DSA projections. Target volume coordinates in the helmet system are calculated using two DSA acquisitions taken with a free angle (approximately 90 degrees), one in the AP and the other in the LL direction. The helmet can be freely positioned between the x-ray source and the image plane. The projections of eight reference points inserted in the helmet at a known location, are used to calculate the transformation matrix between the two coordinate systems. We performed numerical and experimental validation of the system. A hypothetical random error (up to 2 mm) on image coordinates of the reference points allowed to determine that the error in target localization was less than 0.2 mm. Using DSA images of target points with a known location within a phantom, the error between calculated and actual location was, on average, 0.30+/-0.13 mm (mean+/-SD), with a maximum error of 0.49 mm. The results of numerical and experimental validations show that the system we have developed allows fast and accurate localization of the center of the target volume and it is suitable for efficient guiding during stereotactic radiosurgery of AVM. PMID- 11011730 TI - Fast reconstruction with uniform noise properties in halfscan computed tomography. AB - The hybrid algorithms developed recently for the reconstruction of fan-beam images possess computational and noise properties superior to those of the fan beam filtered backprojection (FFBP) algorithm. However, the hybrid algorithms cannot be applied directly to a halfscan fan-beam sinogram because they require knowledge of a fullscan fan-beam sinogram. In this work, we developed halfscan hybrid algorithms for image reconstruction in halfscan computed tomography (CT). Numerical evaluation indicates that the proposed halfscan-hybrid algorithms are computationally more efficient than are the widely used halfscan-FFBP algorithms. Also, the results of quantitative studies demonstrated clearly that the noise levels in images reconstructed by use of the halfscan-hybrid algorithm are generally lower and spatially more uniform than are those in images reconstructed by use of the halfscan-FFBP algorithm. Such reduced and uniform image noise levels may be translated into improvement of the accuracy and precision of lesion detection and parameter estimation in noisy CT images without increasing the radiation dose to the patient. Therefore, the halfscan-hybrid algorithms may have significant implication for image reconstruction in conventional and helical CT. PMID- 11011731 TI - Quantitative in vivo analysis of the kinematics of carpal bones from three dimensional CT images using a deformable surface model and a three-dimensional matching technique. AB - The purpose of this study was to obtain quantitative information of the relative displacements and rotations of the carpal bones during movement of the wrist. Axial helical CT scans were made of the wrists of 11 volunteers. The wrists were imaged in the neutral position with a conventional CT technique, and in 15-20 other postures (flexion-extension, radial-ulnar deviation) with a low-dose technique. A segmentation of the carpal bones was obtained by applying a deformable surface model to the regular-dose scan. Next, each carpal bone, the radius, and ulna in this scan was registered with the corresponding bone in each low-dose scan using a three-dimensional matching technique. A detailed definition of the surfaces of the carpal bones was obtained from the regular-dose scans. The low-dose scans provided sufficient information to obtain an accurate match of each carpal bone with its counterpart in the regular-dose scan. Accurate estimates of the relative positions and orientations of the carpal bones during flexion and deviation were obtained. This quantification will be especially useful when monitoring changes in kinematics before and after operative interventions, like mini-arthrodeses. This technique can also be applied in the quantification of the movement of other bones in the body (e.g., ankle and cortical spine). PMID- 11011732 TI - Evaluation of an algorithm for the assessment of the MTF using an edge method. AB - An algorithm to calculate the presampling modulation transfer function (MTF) of an imaging system from an angled edge image has its own inherent transfer function. Factors such as the angle of the sampling aperture to the edge, registration of edge function profiles using the determined edge angle, differentiation, smoothing, and folding all combine to produce the frequency response of the algorithm. In this work, the profile registration transfer function accounting for an error in the determined edge angle has been derived. This has been incorporated with other, previously reported, algorithm component transfer functions to fully characterize the MTF calculation algorithm. When registering profiles, small errors in the edge angle determination were found to result in large errors in the MTF, as the misalignment errors increase with the number of profiles. For example, registering 50 profiles a 0.07 degree error in a 7 degree edge angle (1% error) produces a 36% error in the MTF at the system cutoff frequency f=f(c) when profiles are oversampled at a frequency f(s)=8f(c)(f(c) is defined as the maximum frequency reproducible without aliasing when sampling at the limiting system Nyquist frequency f(s) = 2f(c)). These results highlight the importance of quantifying the transfer function of the algorithm used to determine an imaging system modulation transfer function. The MTF calculation algorithm and the transfer function analysis have been incorporated into a Windows-based software program to be made available for general use. PMID- 11011733 TI - Analytic reconstruction of magnetic resonance imaging signal obtained from a periodic encoding field. AB - We have proposed a two-dimensional PERiodic-Linear (PERL) magnetic encoding field geometry B(x,y) = g(y)y cos(q(x)x) and a magnetic resonance imaging pulse sequence which incorporates two fields to image a two-dimensional spin density: a standard linear gradient in the x dimension, and the PERL field. Because of its periodicity, the PERL field produces a signal where the phase of the two dimensions is functionally different. The x dimension is encoded linearly, but the y dimension appears as the argument of a sinusoidal phase term. Thus, the time-domain signal and image spin density are not related by a two-dimensional Fourier transform. They are related by a one-dimensional Fourier transform in the x dimension and a new Bessel function integral transform (the PERL transform) in the y dimension. The inverse of the PERL transform provides a reconstruction algorithm for the y dimension of the spin density from the signal space. To date, the inverse transform has been computed numerically by a Bessel function expansion over its basis functions. This numerical solution used a finite sum to approximate an infinite summation and thus introduced a truncation error. This work analytically determines the basis functions for the PERL transform and incorporates them into the reconstruction algorithm. The improved algorithm is demonstrated by (1) direct comparison between the numerically and analytically computed basis functions, and (2) reconstruction of a known spin density. The new solution for the basis functions also lends proof of the system function for the PERL transform under specific conditions. PMID- 11011734 TI - Simultaneous iterative reconstruction for emission and attenuation images in positron emission tomography. AB - The quality of the attenuation correction strongly influences the outcome of the reconstructed emission scan in positron emission tomography. Usually the attenuation correction factors are calculated from the transmission and blank scan and thereafter applied during the reconstruction on the emission data. However, this is not an optimal treatment of the available data, because the emission data themselves contain additional information about attenuation: The optimal treatment must use this information for the determination of the attenuation correction factors. Therefore, our purpose is to investigate a simultaneous emission and attenuation image reconstruction using a maximum likelihood estimator, which takes the attenuation information in the emission data into account. The total maximum likelihood function for emission and transmission is used to derive a one-dimensional Newton-like algorithm for the calculation of the emission and attenuation image. Log-likelihood convergence, mean differences, and the mean of squared differences for the emission image and the attenuation correction factors of a mathematical thorax phantom were determined and compared. As a result we obtain images improved with respect to log likelihood in all cases and with respect to our figures of merit in most cases. We conclude that the simultaneous reconstruction can improve the performance of image reconstruction. PMID- 11011735 TI - Partial wave analysis of the ultrasound comet tail artifact. AB - An adaptation of linear acoustic scattering theory is used to describe the comet tail artifact which is observed in the diagnostic ultrasound imaging of small spherical masses. The calculated backscattered pressure wave is shown to correspond well with the appearance of the artifact in a variety of clinical images. PMID- 11011736 TI - Application of coloring theory to reduce intensity modulated radiotherapy dose calculations. AB - Coloring theory is applied to reduce the dose calculations under intensity modulated radiotherapy. Intensity modulated radiotherapy varies the intensity profile across the beam. The beam face is divided into a panel of small squares or "bixels." Each square may be opened or closed for different lengths of time by moving collimator leaves in and out of the beam. It has been shown that the distribution of dose from radiation directed through any open square depends on whether the adjacent squares are opened or closed. Taking the states of neighboring bixels into account greatly increases the required dose calculations. There are 2(8) possible ways to select open or closed states for the eight neighbors of a given bixel. Each combination represents one coloring of a 3 x 3 panel, and each coloring demands a separate dose calculation. The number of calculations is reduced by considering the symmetries of a square. The 256 possible colorings can be divided into 51 distinct patterns through application of Burnside's theorem. Each pattern consists of selections of closed bixels that are the same except for a symmetric transformation of coordinates. If the symmetry between x and y coordinates is broken by collimator leaves whose ends and sides have different effects on bordering bixels, the number of patterns increases to 84. The theoretic gain in the number of calculations through the application of Burnside's theorem is fivefold if bixel borders are symmetric, and threefold if the borders are asymmetric. The results are applied to examples of generated intensity maps. The symmetry rules divide the bixel arrangements into proportionately fewer patterns as the intensity maps become larger, allowing computational gains to be achieved. PMID- 11011737 TI - Computer verification of fluence map for intensity modulated radiation therapy. AB - In a treatment planning system for intensity modulated radiation therapy (IMRT), the time sequence of multileaf collimator (MLC) settings are derived from an optimal fluence map as a postoptimization process using a software module called a "leaf sequencer." The dosimetric accuracy of the dynamic delivery depends on the functionality of the module and it is important to verify independently the correctness of the leaf sequences for each field of a patient treatment. This verification is unique to the IMRT treatment and has been done using radiographic film, electronic portal imaging device (EPID) or electronic imaging system (BIS). The measurement tests both the leaf sequencer and the dynamic multileaf collimator (MLC) delivery system, providing a reliable assurance of clinical IMRT treatment. However, this process is labor intensive and time consuming. In this paper, we propose to separate quality assurance (QA) of the leaf sequencer from the dynamic MLC delivery system. We describe a simple computer algorithm for the verification of the leaf sequences. The software reads in the leaf sequences and simulates the motion of the MLC leaves. The generated fluence map is then compared quantitatively with the reference map from the treatment planning system. A set of pre-defined QA indices is introduced to measure the "closeness" between the computed and the reference maps. The approach has been used to validate the CORVUS (NOMOS Co., Sewickley, PA) treatment plans. The results indicate that the proposed approach is robust and suitable to support the complex IMRT QA process. PMID- 11011738 TI - An optimized forward-planning technique for intensity modulated radiation therapy. AB - Intensity modulated radiation therapy (IMRT) has stirred considerable excitement in the radiation oncology community. Its objective is to make the dose conform to the tumor and spare other organs. Instead of resorting to the rather complex inverse-planning, the technique described here is an extension of the conventional treatment planning technique. The beam orientation and wedge angles are chosen in the conventional rule-based manner. However, within each conformal beam's eye view (BEV) field including margin, a number of sub-field openings are added. The smaller field openings are designed to irradiate the tumor, while sparing the normal tissue of the organs at risk (OARs) that intrude into the target region in the BEV. As the number of intrusions into the target BEV increases, the number of sub-fields for each beam increases. The Cimmino simultaneous projection method was employed to obtain the optimized weighting for each field of each beam. In cases where the dose constraints for the tumor and for the OARs are reasonable, it is possible to obtain a plan with a fairly small number of beams that satisfies the specified dose objectives. This is illustrated for the treatment of prostate cancer, where the rectum creates a concavity in the planning target volume. An advantage of this technique is that the quality assurance for the delivery of these plans does not require extensive special efforts. PMID- 11011739 TI - Estimating the dose variation in a volume of interest with explicit consideration of patient geometric variation. AB - A method to measure the effects of internal organ motion and deformation and patient setup error on cumulative dose variation in a volume of interest is proposed. The method uses multiple CT scans and electronic portal images of a single patient to numerically simulate dose-volume effects over the entire course of the patient's external beam treatment. The results are expressed in the form of a novel dose-volume histogram, called an expected dose-volume histogram (EDVH). PMID- 11011740 TI - A two-step algorithm for predicting portal dose images in arbitrary detectors. AB - Recently, portal imaging systems have been successfully demonstrated in dosimetric treatment verification applications, where measured and predicted images are quantitatively compared. To advance this approach to dosimetric verification, a two-step model which predicts dose deposition in arbitrary portal image detectors is presented. The algorithm requires patient CT data, source detector distance, and knowledge of the incident beam fluence. The first step predicts the fluence entering a portal imaging detector located behind the patient. Primary fluence is obtained through ray-tracing techniques, while scatter fluence prediction requires a library of Monte Carlo-generated scatter fluence kernels. These kernels allow prediction of basic radiation transport parameters characterizing the scattered photons, including fluence and mean energy. The second step of the algorithm involves a superposition of Monte Carlo generated pencil beam kernels, describing dose deposition in a specific detector, with the predicted incident fluence. This process is performed separately for primary and scatter fluence, and yields a predicted dose image. A small but noticeable improvement in prediction is obtained by explicitly modeling the off axis energy spectrum softening due to the flattening filter. The algorithm is tested on a slab phantom and a simple lung phantom (6 MV). Furthermore, an anthropomorphic phantom is utilized for a simulated lung treatment (6 MV), and simulated pelvis treatment (23 MV). Data were collected over a range of air gaps (10-80 cm). Detectors incorporating both low and high atomic number buildup are used to measure portal image profiles. Agreement between predicted and measured portal dose is better than 3% in areas of low dose gradient (<30%/cm) for all phantoms, air gaps, beam energies, and detector configurations tested here. It is concluded that this portal dose prediction algorithm is fast, accurate, allows separation of primary and scatter dose, and can model arbitrary detectors. PMID- 11011741 TI - Dependence of the tray transmission factor on collimator setting and source surface distance. AB - When blocks are placed on a tray in megavoltage x-ray beams, generally a single correction factor for the attenuation by the tray is applied for each photon beam quality. In this approach, the tray transmission factor is assumed to be independent of field size and source-surface distance (SSD). Analysis of a set of measurements performed in beams of 13 different linear accelerators demonstrates that there is, however, a slight variation of the tray transmission factor with field size and SSD. The tray factor changes about 1.5% for collimator settings varying between 4x4 cm and 40 x 40 cm for a 1 cm thick PMMA tray and approximately 3% for a 2 cm thick PMMA tray. The variation with field size is smaller if the source-surface distance is increased. The dependence on the collimator setting is not different, within the experimental uncertainty of about 0.5% (1 s.d.), for the nominal accelerating potentials and accelerator types applied in this study. It is shown that the variation of the tray transmission factor with field size and source-surface distance can easily be taken into account in the dose calculation by considering the volume of the irradiated tray material and the position of the tray in the beam. A relation is presented which can be used to calculate the numerical value of the tray transmission factor directly. These calculated values can be checked with only a few measurements using a cylindrical beam coaxial miniphantom. PMID- 11011742 TI - A noninvasive dose estimation system for clinical BNCT based on PG-SPECT- conceptual study and fundamental experiments using HPGe and CdTe semiconductor detectors. AB - A noninvasive method for measuring the absorbed dose distribution during the administration of clinical boron neutron capture therapy (BNCT) using an online three-dimensional (3D) imaging system is presented. This system is designed to provide more accurate information for treatment planning and dosimetry. The single-photon emission computed tomography (SPECT) technique is combined with prompt gamma-ray analysis (PGA) to provide an ideal dose estimation system for BNCT. This system is termed PG-SPECT. The fundamental feasibility of the PG-SPECT system for BNCT is confirmed under the following conditions: (1) a voxel size of 1 x 1 x 1 cm3, comparable to the spatial resolution of our standard dosimetric technique using gold wire activation, where data are available for every 5-10 mm of wire length; (2) a reaction rate of 10B(n,alpha)7Li within the measurement volume is greater than 1.1 x l0(6) interactions/cm3/s, corresponding to a thermal neutron flux of 5 x 10(8) n/cm2/s and a 10B concentration of greater than 10 ppm for the deepest part of the tumor volume under typical BNCT clinical conditions; (3) statistical uncertainty of the count rate for 10B(n,alpha)7Li prompt gamma rays is 10% or less. The desirable characteristics of a detector for the PG-SPECT system were determined by basic experiments using both HPGe and CdTe semiconductor detectors. The CdTe semiconductor detector has the greatest potential for this system because of its compactness and simplicity of maintenance. PMID- 11011743 TI - Proton loss model for therapeutic beam dose calculations. AB - A transport algorithm called the proton loss (PL) model is developed for proton pencil beams of therapeutic energies. The PL model takes into account inelastic nuclear reactions, pathlength straggling, and energy-loss straggling and predicts the 3D dose distribution from a proton pencil beam. In proton beams, the multiple scattering and ionizational energy loss processes approach their diffusional limit where scattering and energy loss probability densities become Gaussian. Therefore we chose to derive the PL model from the Fermi-Eyges diffusional multiple scattering theory and the Gaussian theory of energy straggling. We first introduce a generalization of the Fermi-Eyges equation for proton pencil beams, labeled the proton loss (PL) transport equation. This new equation includes terms that model inelastic nuclear reactions as a depth-dependent absorption and pathlength straggling as a quasi-absorption. Then energy straggling is taken into account by using a weighted superposition of a discrete number of elementary pencil beams. These elementary pencil beams have different initial energies and lose energy according to the CSDA, thus they have different ranges of penetration. A final solution for the proton beam transport is obtained as a linear combination of elementary pencil beam solutions with weights defined by the Gaussian evolution of the proton energy spectrum with depth. A numerical comparison of the dose distribution predictions of the PL model with measurements and PTRAN Monte Carlo simulations indicates the model is both computational fast and accurate. PMID- 11011744 TI - GammaPlan-Leksell Gamma Knife radiosurgery treatment planning verification method. AB - This work provides a method for an independent check of Gamma Knife GammaPlan radiosurgery calculations, named the spherical approximation method or SAM. Based on skull dimension measurements, the treated volume of the head is modeled as a sphere of radius R. With this approximation, an analytical solution for fast ray tracing of the path length, for each of the 201 beamlets, of the Gamma Knife helmet collimator was possible. The dose rate at the focus of a single shot is the sum of the contributions of all active beamlets adjusted for both the collimator factor and attenuation. For an arbitrary point, the dose rate is derived at the beamlet level from the focus values adjusted for the new path length attenuation and the appropriate collimators' off-axis profiles. The sum over all beamlets' contributions gives the dose rate at that particular point. At the single shot level, SAM independent check results agree with the GammaPlan for patient calculations to better than +/-6% and, as expected, in spherical phantoms the agreements improve to better than +/-1.0%. For an arbitrary point, multi-shot procedure, the agreement is better than +/-3% and +/-1.5, respectively. PMID- 11011745 TI - Red marrow dosimetry for radiolabeled antibodies that bind to marrow, bone, or blood components. AB - Hematologic toxicity limits the radioactivity that may be administered for radiolabeled antibody therapy. This work examines approaches for obtaining biodistribution data and performing dosimetry when the administered antibody is known to bind to a cellular component of blood, bone, or marrow. Marrow dosimetry in this case is more difficult because the kinetics of antibody clearance from the blood cannot be related to the marrow. Several approaches for obtaining antibody kinetics in the marrow are examined and evaluated. The absorbed fractions and S factors that should be used in performing marrow dosimetry are also examined and the effect of including greater anatomical detail is considered. The radiobiology of the red marrow is briefly reviewed. Recommendations for performing marrow dosimetry when the antibody binds to the marrow are provided. PMID- 11011746 TI - Accuracy in catheter reconstruction in computed tomography planning of high dose rate prostate brachytherapy. AB - In high dose rate prostate brachytherapy, inadequate reconstruction of catheter geometry in treatment planning may result in erroneous dose delivery. Catheters may be digitized with: (1) Parallel reconstruction: digitized at only one point and assumed parallel and horizontal: (2) Straight reconstruction: digitized at both ends and assumed straight while at an angle: (3) Slice-by-slice reconstruction: digitized on all slices to obtain exact geometry. Our results show that individual catheters are often not parallel to each other, but fairly straight. Parallel reconstruction is the least accurate for dosimetric planning, while slice-by-slice reconstruction is time-consuming. Straight (two-point) reconstruction represents a balance between accuracy and efficiency. PMID- 11011747 TI - Experimental determination of dosimetric characteristics of Best 125I brachytherapy source. AB - 125I brachytherapy sources are being used for interstitial implants in tumor sites such as the prostate. Recently, the Best 125I source became commercially available for interstitial brachytherapy treatment. Dosimetric characteristics (dose rate constant, radial dose function, and anisotropy function) of this source were experimentally determined, following the AAPM Task Group 43 recommendations, and were related to the NIST 1999 calibration assigned to this source. Measurements were performed in Solid Water phantom using LiF thermoluminescent dosimeters. The results indicated a dose rate constant, lambda, of 1.01 +/- 0.08 cGy h(-1) U(-1) for the new source. The radial dose function, g(r), of the new source was measured at distances ranging from 0.5 to 10.0 cm. The anisotropy function, F(r, theta), of the new source was measured at distances of 2, 5, and 7 cm from the source center. These data compare favorably with those from the Nycomed/Amersham Models 6711 and 6702 sources. The anisotropy constant, phi(an), of the Best 125I source was found to be 0.982. Complete dosimetric parameters of the new source are presented in this paper. PMID- 11011748 TI - Dosimetric characteristics of the Pharma Seed model BT-125-I source. AB - 125I brachytherapy sources are being used with increasing frequency for interstitial implants in tumor sites, especially the prostate. Recently, a new 125I source design has become commercially available for clinical applications. Dosimetric characteristics (i.e., dose rate constant, radial dose function, and anisotropy function) of this source were experimentally and theoretically determined following the AAPM Task Group 43 (TG-43) recommendations and were related to the 1999 NIST calibration assigned to this source [S(k), 99std]. Measurements were performed in a Solid Water phantom using LiF thermoluminescent dosimeters. The measured data were used to validate the Monte Carlo simulations that were performed in Solid Water using the PTRAN code. The Monte Carlo calculations were then performed in liquid water to obtain the dosimetric information for clinical applications in accordance with TG-43 recommendations. The results indicated that the dose rate constant, lambda, of the Pharma Seed model BT-125-I 125I source was 0.90 +/- 0.06 cGy h(-1) U(-1) using thermoluminescent dosimeter (TLD) measurements and 0.92 +/- 0.03 cGy h(-1) U(-1) using Monte Carlo simulations in Solid Water. The calculated value in liquid water was found to be 0.95 +/- 0.03 cGy h(-1) U(-1). The radial dose function, g(r), of the new 125I source was measured at distances ranging from 0.5 to 10 cm using LiF TLD in Solid Water phantom material. The Monte Carlo simulations were performed for distances ranging from 0.1 to 10 cm from the source center in Solid Water and liquid water. The anisotropy function, F(r, theta), was measured at distances of 2, 5, and 7 cm from the source center and calculated at distances of 0.5, 1, 2, 3, 5, and 7 cm from the source center. The anisotropy constant, phi(an), of the Pharma Seed source in water was found to be 0.975. Complete dosimetric data are described in this manuscript. Per TG-43, the values reported in water should be used for clinical treatment planning systems. PMID- 11011749 TI - A Monte Carlo investigation of the dosimetric characteristics of the CSM11 137Cs source from CIS. AB - The purpose of this study is to calculate basic dosimetry data for a CSM11 low dose rate 137Cs source in water. This source is widely used in afterloadable dome cylinders designed to homogeneously irradiate the vaginal cuff alone or additional areas of the vagina in hysterectomized patients. In this study, the Monte Carlo simulation code GEANT, incorporating in detail source geometry, is used to investigate the dosimetric characteristics of the source. The calculated data were analyzed using a fitting procedure that is described in detail. Absolute dose rate distributions in water were calculated around this source and are presented as conventional 2D Cartesian lookup tables (classically along-away tables). Also, the dose calculation formalism endorsed by the Interstitial Collaborative Working Group and the AAPM Task Group 43 have been calculated. The calculated dose rate constant for this source is lambda = 1.096 +/- 0.002 cGy h( 1) U(-1). The anisotropy function results in about 50% deviations from isotropy at positions near the long axis of the source. The radial dose function is given as a polynomial that reproduces the calculated data up to 20 cm. Best-fit values of attenuation coefficients suitable for use in Sievert integral calculations have been derived. PMID- 11011750 TI - Comment on "In vivo biodistribution of 125IPIP and internal dosimetry of 123IPIP radioiodinated agents selective to the muscarinic acetylcholinergic receptor complex" [Med. Phys. 27, 778-786 (2000)]. PMID- 11011751 TI - Comment on "Full breast digital mammography with an amorphous silicon-based flat panel detector: physical characteristics of a clinical prototype" [Med. Phys. 27, 558-567 (2000)]. PMID- 11011752 TI - Management of asthma exacerbations: home treatment. AB - Asthma is a major cause of morbidity in children. Delays in care and inappropriate home management practices can contribute to morbidity and mortality. Guidelines for the diagnosis and management of asthma were developed in 1991 and revised in 1997 by The National Heart, Lung and Blood Institute. In this article we review the recommended pharmacological protocol for home treatment of asthma exacerbations, and then discuss in more detail behavioral components of asthma management, including monitoring of symptoms, seeking medical care, developing clinician-patient partnerships, and practical issues in equipment and medication usage. Discrepancies between guideline recommendations and current management practices are also discussed. PMID- 11011753 TI - Applicability of the Asthma Opinion Survey in the Spanish population: distribution and relationship with sociodemographic and clinical variables. AB - We evaluated a Spanish version of the Asthma Opinion Survey (AOS) in a sample of 90 adult Spanish asthma patients. The Spanish AOS showed acceptable internal consistency, and a factorial structure similar to that reported by others, i.e., vulnerability, perceived quality of care, and recognition and control, although we additional ly obtained a fourth factor, attitudes toward patient knowledge. Vulnerability was significantly higher among older patients, and among patients with lower educational level, lower income, and more severe asthma. Perceived quality of care, and recognition and control were positively correlated with frequency of attacks, and recognition and control was significantly higher in older patients. The factor attitudes toward patient knowledge was significantly higher in younger patients and in patients with relatively high income. These results support the cross-cultural validity of this questionnaire. PMID- 11011754 TI - The association of skin test reactivity, total serum IgE levels, and peripheral blood eosinophilia with asthma in Kuwait. AB - There is evidence that elevated serum immunoglobulin E (IgE) and eosinophilia correlate well with allergic skin test reactivity. These parameters have been used as alternative methods to characterize atopic subjects. Skin test reactivity is the only measure used routinely in clinical practice in Kuwait to reflect atopy in asthma patients. This study examines the usefulness of the two other parameters of atopy in patients with asthma, and to determine the most common allergens involved in Kuwait. Between 1998 and 1999, 101 asthma patients and 33 healthy controls were recruited for this study. Skin sensitivity test, serum total and specific IgE, total blood eosinophil count (B-EOS), and eosinophil cationic protein (ECP) tests were performed in patients and controls. Nine allergens known to be prevalent in this environment were selected for the skin test and specific IgE test. Spirometry was also measured. These parameters were repeated after 4 weeks of therapy in the patients only. Skin test reaction was positive in 81% of the patients, while total IgE above 200 kU/L was obtained in 63% of cases. B-EOS above 300 x 10(3)/L was found in 75% of cases. House dust mite reactivity (positivity) was the most frequently encountered skin allergy, occurring in 28% of the patients. IgE correlated positively with B-EOS and ECP. B EOS similarly correlated positively with ECP. There was a negative correlation between ECP and forced expiratory volume in 1 sec (FEV1) (% predicted) as expected. At least one positive parameter of atopy was found in 95% of the patients. In 48% of the patients, all three parameters of atopy were found to be positive. Skin test reactivity and elevated IgE were found together in 62% of the cases. This study reveals a significant degree of allergy among patients with asthma in this environment. Skin testing was found to be the most effective measure of atopy in this environment, and correlates well with the other more sensitive newer tests. PMID- 11011755 TI - Assessment of magnesium status in patients with bronchial asthma. AB - To elucidate the contribution of magnesium to bronchial hyperreactivity in patients with stable bronchial asthma, magnesium concentrations in serum (S-Mg), erythrocytes (R-Mg), and lymphocytes (L-Mg) were measured in 25 patients with bronchial asthma (BA group) and 9 age-matched healthy subjects (control group). A parenteral magnesium loading test, a continuous low-dose magnesium infusion of 0.2 mEq/kg over 4 hr, was performed in 10 of 25 asthmatic patients and in the control group. R-Mg was significantly lower in the BA group than in the control group (4.96 +/- 0.47, 6.13 +/- 0.62 mEq/L, p < 0.001, respectively), although S Mg (2.4 +/- 0.1, 2.4 +/- 0.2 mg/dL) and L-Mg (1.28 +/- 0.26, 1.15 +/- 0.13 microg/mg/protein) were not significantly different between the two groups. Magnesium deficiency in total body stores was revealed in 40% of patients (4/10 patients) and 11% of control subjects (1/9 subjects) by parenteral magnesium loading test. The ratio of magnesium retention to urinary excretion through the parenteral magnesium loading test showed a significant inverse correlation with R Mg (r = -0.78, p < 0.01). Bronchial reactivity to inhaled methacholine had a significant inverse correlation with R-Mg (r = -0.42, p < 0.05). We conclude that 40% of asthmatic patients demonstrated magnesium deficiency, and that the low magnesium concentration in erythrocytes reflects decreased magnesium stores in patients with bronchial asthma. PMID- 11011756 TI - Secretion of adrenocorticotropic hormone induced by allergen inhalation in patients with atopic asthma. AB - Allergen inhalation in atopic patients results in cytokines production or release of preformed cytokines, some of which are known to induce adrenocorticotropic hormone (ACTH) secretion in experimental conditions. We examined whether allergen inhalation can induce ACTH secretion in vivo. A significant elevation of ACTH levels was observed in 2 and 24 hr after allergen inhalation challenge. However, methacholine challenge with the same degree of airflow limitation did not induce ACTH elevation, indicating that this may not be due to bronchoconstriction per se. Our results indicate that allergen inhalation can trigger ACTH secretion in patients with atopic asthma. PMID- 11011757 TI - A study of the relationship among self-reported noncompliance, symptomatology, and psychological variables in patients with asthma. AB - We examined the association among self-reported noncompliance and clinical status, symptomatology, functional activity, and psychological variables in asthma. Eighty-five asthmatics, who were hospitalized (H group), outpatients previously hospitalized (OH group), or outpatients not previously hospitalized (O group) completed questionnaires. Compliance was assessed during a structured interview. The higher prevalence of self-reported noncompliance in H group and OH group, compared to O group, was not explained by differences in respiratory function. Self-reported noncompliance was related to symptoms and emotional distress associated with disease and treatment, but not to functional or emotional status. A patient subgroup that catalogued itself as noncompliant may also be at risk for hospitalization, and was characterized by emotional distress associated with disease and treatment. PMID- 11011758 TI - Inhaled corticosteroid nonadherence and immediate avoidable medical events in older adults with chronic pulmonary ailments. AB - This study examined the association between nonadherence to inhaled corticosteroids and risk of an immediate avoidable medical event (emergency department visit and hospitalization) in elderly patients with chronic pulmonary ailments. The study comprised a cross-sectional analysis comparing patients with and without avoidable medical event occurrence in each year for adherence preceding the event, and a case-crossover analysis examining the same patient's adherence in years with and without events. The cross-sectional study showed risk odds reductions of nearly 20% in the partially adherent patient, and 40% in the adherent patient compared to the nonadherent patient, after controlling for potential group differences. PMID- 11011759 TI - Knowledge, attitudes, and practices of parents of asthmatic children in Cape Town. AB - To obtain information useful to asthma care in a relatively poor, high asthma prevalence population, a focus group study was undertaken with 72 parents of children with asthma or recurrent wheezing. There was a reasonable level of understanding of the causes of asthma, although variable acceptance of the diagnosis. Willingness to undertake home management of acute episodes and environmental control measures was high. Reported treatment was characterized by reliance on syrups, use of home remedies, resistance to inhaled therapy, and relatively low compliance with prescribed treatment. There was agreement on the poor level of service offered by doctors and public sector clinics. Asthma education in this population needs to build on what parents know and accept, but stress the relative safety of inhaled therapy, the need for maintenance therapy, and the value of pre-exercise prophylaxis, house dust mite control, and smoking cessation. There is an urgent need to improve the quality of asthma care provided by public sector clinics. PMID- 11011760 TI - Stemming the tide of genomics: computational genomics III. PMID- 11011761 TI - Relationship between whole proteome aminoacid composition and static DNA curvature. AB - To study possible relationships between an organism's genomic DNA curvature and the aminoacid composition of its proteome, every peptidic sequence from fully determined genomes was retrotranslated using the E. coli codon preferences, and the curvature profiles of the resulting DNA sequences were calculated and compared. A clear interdependence between these two variables was observed, as each retrotranslated proteome presented a distinctive, statistically significant DNA curvature profile biased toward its natural DNA curvature profile. In addition, by comparing the profiles arising from real and randomly permuted proteomes, we also found a position-dependent contribution of the peptidic sequence to DNA curvature. The implications of these results support the idea of a possible selection toward a specific global curvature of genomes. PMID- 11011762 TI - Improving quality of expressed sequence tag (EST) databases: recovery of reversed, antisense cDNA sequences. AB - Expressed sequence tag (EST) databases contain a significant number (5-20%) of reversed, antisense, cDNA sequences that can be recognized by the label "reversed clone: similarity on wrong strand" in the annotations to the sequence. Despite this high number of altered sequences, no attempt has been made to explain the alteration in molecular terms, or to evaluate their effect on the quality of the information curated in EST databases. In this paper we try to explain the way these altered sequences are originated, and propose a plausible mechanism: a "double priming" of the first strand oligo-dT primer at both ends of nascent cDNAs. In this way, a symmetrical cDNA intermediate is generated, an intermediate that can be cloned after partial digestion with the restriction enzyme used for the directional cloning. Furthermore, when "secondary" priming takes place inside the cDNA, the chain synthesized is prone to be truncated prematurely, with the subsequent loss of upstream information. One of the most subtle effects of this cloning alteration is the generation of virtual open reading frames (ORFs) in sequences with no homologues available for comparison. Nevertheless, and according to our model and our data, the "double priming mechanism" does not shift the ORF effected, so antisense sequences should be considered as normal ones after a simple transformation in their inverse-complementary forms. PMID- 11011763 TI - Sequencing of the Francisella tularensis strain Schu 4 genome reveals the shikimate and purine metabolic pathways, targets for the construction of a rationally attenuated auxotrophic vaccine. AB - Francisella tularensis is the etiological agent of tularemia, a serious disease in several Northern hemisphere countries. The organism has fastidious growth requirements and is very poorly understood at the genetic and molecular levels. Given the lack of data on this organism, we undertook the sample sequencing of its genome. A random library of DNA fragments from a highly virulent strain (Schu 4) of F. tularensis was constructed and the nucleotide sequences of 13,904 cloned fragments were determined and assembled into 353 contigs. A total of 1.83 Mb of nucleotide sequence was obtained that had a G+C content of 33.2%. Genes located on plasmids pOM1 and pNFL10, which had been previously isolated from low virulence strains of F. tularensis, were absent but all of the other known F. tularensis genes were represented in the assembled data. F. tularensis Schu4 was able to grow in the absence of aromatic amino acids and orthologues of genes which could encode enzymes in the shikimate pathway in other bacteria were identified in the assembled data. Genes that could encode all of the enzymes in the purine biosynthetic and most of the en- zymes in the purine salvage pathways were also identified. This data will be used to develop defined rationally attenuated mutants of F. tularensis, which could be used as replacements for the existing genetically undefined live vaccine strain. PMID- 11011764 TI - Molecular cloning of the Atlantic cod chymotrypsinogen B. AB - The cDNA encoding Atlantic cod (Gadus morhua) chymotrypsinogen B has been isolated and sequenced. Its deduced amino acid sequence consists of a 16-residue signal sequence and a mature polypeptide of 247 residues, being two residues longer than its vertebrate analogs. This mature polypeptide corresponds to a calculated molecular mass of 26.5 kDa and shares 70% sequence identity with cod chymotrypsinogen A. However, the identity between cod chymotrypsinogen B and its other vertebrate analogues is 63-66%. In common with most fish serine proteases, cod chymotrypsinogen B contains a high number of methionine residues. The presence of a threonine instead of a highly conserved serine residue at position 189 is a novel characteristic of this enzyme. Cod chymotrypsin B, as its type B vertebrate analogs, has an alanine at position 226, whereas a glycine is most commonly found at this position in the type A chymotrypsins. PMID- 11011765 TI - Causal analysis of CpG suppression in the Mycoplasma genome. AB - Some bacterial genomes are known to have low CpG dinucleotide frequencies. While their causes are not clearly understood, the frequency of CpG is suppressed significantly in the genome of Mycoplasma genitalium, but not in that of Mycoplasma pneumoniae. We compared orthologous gene pairs of the two closely related species to analyze CpG substitution patterns between these two genomes. We also divided genome sequences into three regions: protein-coding, noncoding, and RNA-coding, and obtained the CpG frequencies for each region for each organism. It was found that the observed/expected ratio of CpG dinucleotides is low in both the protein-coding and noncoding regions; while that ratio is in the normal range in the RNA-coding region. Our results indicate that CpG suppression of the Mycoplasma genome is not caused by (1) biased usage amino acid; (2) biased usage of synonymous codon; or (3) methylation effects by the CpG methyltransferase in the genomes of their hosts. Instead, we consider it likely that a certain global pressure, such as genome-wide pressure for the advantages of DNA stability or replication, has the effect of decreasing CpG over the entire genome, which, in turn, resulted in the biased codon usage. PMID- 11011766 TI - Probing biopolymers with the atomic force microscope: a review. AB - This short review presents an overview of atomic force microscopy (AFM) of biopolymers and specific examples of some of the biopolymers that have been analyzed by AFM. These specific examples include extracellular polymeric substances on the surfaces of bacterial biofilms, condensed DNA, DNA constructs, and DNA-protein interactions. In addition, two examples are presented for AFM analyses of proteins: laminin flexing its arms in solution and neurofilaments entropically brushing away the space around themselves. PMID- 11011767 TI - Biocompatibility and performance in vitro of a hemostatic gelatin sponge. AB - The biocompatibility of a hemostatic gelatin sponge (Cutanplast Standard) was evaluated in vitro. Cytotoxicity was assessed by neutral red uptake and amido black staining tests; genotoxicity was assayed using the Ames test, Sister Chromatides Exchanges (SCE) and chromosomal aberrations. The ability of the hemostatic gelatin sponge to induce platelet adhesion and release reaction was also determined. The undiluted extract of the test material was found to be cytotoxic, but cell viability was not affected by 1:2 and 1:10 diluted extract. The same extract was found to be non-genotoxic using the three assays for genotoxicity. A significant decrease of platelet number, as well as a significant release of platelet factor 4 was found to be caused by the solid material. In conclusion, Cutanplast Standard is neither cytotoxic nor genotoxic, while inducing platelet adhesion and release reaction when challenged with blood. PMID- 11011768 TI - Surface characterization and platelet adhesion studies on fluorocarbons prepared by plasma-induced graft polymerization. AB - It is believed that the interactions between the biological environment and biomaterial surface are the key factors influencing its biocompatibility. Therefore, plasma processing, which can vary the surface properties without altering the bulk properties, has been considered as one of the important techniques for improving a materials' biocompatibility. In this investigation, plasma-induced grafting polymerization of vinylidene fluoride (VDF) and chlorotrifluoroethylene (CTFE), instead of direct plasma polymerization, was attempted with an aim to improve the substrate blood compatibility. Contact angle measurement indicated both fluorocarbon-grafted Pdyethylenes (PEs) are hydrophobic. Due to the additional fluorine and chlorine atoms on the CTFE chain, the PCTFE-grafted PE exhibited a higher hydrophobicity than the PVDF-grafted one. ESCA analysis has revealed that these two plasma-induced fluorocarbon deposits contain almost no CFx (x > 2) binding on the surface layer, indicating the grafting polymerization mainly follows the free radical mechanism instead of the molecule-highly-fragmented reaction steps commonly seen in the direct plasma polymerization treatment. In addition, ATR-FTIR has shown the surface chemical configuration of these PVDF- and PCTFE-grafted PEs to be very similar to those of the bulk samples of PVDF and PCTFE. The surface roughness decreased after oxygen plasma treatment and was further reduced by VDF and CTFE grafting polymerization. In vitro platelet adhesion testing indicated these two fluorocarbon grafted PEs are less platelet-activating than the nontreated PE control and oxygen plasma activated one. PMID- 11011769 TI - Extended release peptide delivery systems through the use of PLGA microsphere combinations. AB - The purpose of this study was to evaluate the utility of combining polymer matrices to overcome extended lag periods or unacceptably short durations of action intrinsic in the individual polymer systems. Leuprolide, an LHRH superagonist, was incorporated into a variety of poly(lactide-co-glycolide) (PLGA) matrices using a solvent extraction/evaporation method. The in vitro release of Leuprolide from these matrices was evaluated at pH 7.0 and 37 degrees C in phosphate buffer. The formulations were administered to an animal model at 3 or 9 mg kg(-1) doses and serum testosterone levels were followed using a RIA method. A two-part system was made by combining microspheres made from a 75:25 acid terminated PLGA and microspheres made from a 75:25 ester terminated PLGA. This combination elicited chemical castration from 10-100 days. A three-part combination composed of an ester terminated 75:25 PLGA formulation, an ester terminated 50:50 PLGA formulation and an acid terminated 50:50 PLGA formulation also provided a composite profile with an onset of 10 days and a duration of approximately 100 days. Additionally, a single polymer system composed of a high molecular weight ester terminated 75:25 PLGA was employed to produce release over the desired 90-day release period. This study demonstrates that microsphere combinations can potentially provide effective therapies over extended intervals when combined at the proper ratio. PMID- 11011770 TI - Intensive promotion of spheroid formation by soluble factors in a hepatocyte conditioned medium. AB - We developed a hybrid artificial liver and a drug metabolism simulator using polyurethane foam (PUF) in which primary hepatocytes spontaneously form functional spheroids. Gel filtration liquid chromatography analysis of a hepatocyte-conditioned medium during spheroid formation showed that some substances secreted by primary rat hepatocytes accumulated advantageously inside the pores of PUF compared with outside. Similar substances were detected in a hepatocyte-conditioned medium from a positively-charged surface by concentrating the substances using an ultrafiltration membrane of a molecular weight-cutoff of 50 kD. These substances were shown to act as soluble factors on freshly isolated primary rat hepatocytes to promote spontaneous and rapid spheroid formation, depending on their concentration by preventing them from initially attaching and spreading on a positively-charged surface. In particular, using 50-fold concentrated substances, about 80% of total hepatocytes formed the floating spheroids within 72 h of culture. The resulting spheroids had a diameter distribution mainly ranging from 40 to 70 microm and expressed high-level liver specific functions compared with a conventional monolayer. PMID- 11011771 TI - Self-complex formation of nicotinamide-modified dextran with carboxymethyl dextran using their degradation products. AB - A pseudo-metabolic cycle as a self-degradation system was designed: enzymatic degradation products from a polysaccharide generate oxidants which introduce a cationic charge into the polysaccharide chains, and can form a polyion complex with an anionic polysaccharide. As a component of such a system, dextran, with various degrees of nicotinamide substitution, was prepared. Its degradation by dextranase, redox reaction via glucose oxidase-catalysis, and polyion complex formation with carboxymetyl dextran (CMD) were examined. Nicotinamide-modified dextran (NA-Dex) with nine nicotinamide moieties per 100 glucose units was soluble in PBS and completely oxidized by > 100 mM H2O2. The oxidized type of NA Dex was found to form a 1:1 complex with CMD. By the addition of dextranase, isomaltase, and glucose oxidase (GOD) to phosphate buffer solution of the reduced type of NA-Dex and CMD, the transmittance of the solution dropped, suggesting polyion complex formation via the oxidation of 1,4-dihydronicotinamide in NA-Dex by H2O2 generated from GOD-catalytic reaction. These findings are of great importance for designing a self-complex formation system aimed at biodegradable and osillative drug release. PMID- 11011772 TI - Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata. AB - A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEK-OH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[] NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN. PMID- 11011773 TI - Retrograde nailing for subtrochanteric femur fractures. AB - This study reviews the results of subtrochanteric femur fractures treated with a retrograde nail at a level 1 trauma center. Sixteen patients with 17 fractures were followed up until union. The average Injury Severity Score (ISS) was 19, and average knee range of motion was 127 degrees. Four patients required dynamization of their nail, and three healed over a prolonged period of time. Two patients required exchange nailing: one after failure of union after dynamization and one with a broken nail that was revised to an antegrade nailing. The average varus deformity was 5.06 degrees and only 35% of fractures healed in greater than 5 degrees of varus, with an average time to union of 22.4 weeks (range, 6-54 weeks). Antegrade nailing remains the treatment of choice for the vast majority of subtrochanteric femur fractures. However, retrograde femoral nailing may be an effective treatment option for some subtrochanteric fractures in a selected group of patients. PMID- 11011774 TI - The destroyed elbow. AB - Surgeons need an advanced knowledge of upper extremity anatomy and function to care for the destroyed elbow. This paper describes strategies for evaluating and treating this severe injury. Outcomes can be optimized with contemporary techniques and technologies. Surgeons need to instill realistic expectations in patients with advanced compromise. PMID- 11011775 TI - Transsacral versus modified pelvic landmarks for percutaneous iliosacral screw placement--a computed tomographic analysis and cadaveric study. AB - The alar roots of the first sacral body are the usual confines for iliosacral screw (IS) placement when stabilizing a sacroiliac joint injury or sacral fracture. The traditional transsacral method of IS placement aligns the screw horizontally through the sacral ala on both the inlet and outlet views of the sacrum. A modified oblique method of IS placement aligns the screw in an oblique fashion, directed inferiorly to superiorly and posteriorly to anteriorly. The purpose of this investigation was to first define the S-1 segment boundaries for both methods of placement by analyzing the 3-dimensional (3-D) composites of 40 pelvic computed tomography (CT) scans, and then to evaluate the actual placement of ISs under fluoroscopy in 10 cadaveric pelves comparing the transsacral with the modified oblique techniques. Critical dimensions of 7.3 mm and 14.6 mm were considered as the diameter sizes of one and two cannulated screws, respectively. From the 3-D CT composites, the mean anterior/posterior (A/P) measurements were 10.9 mm and 18.0 mm, comparing transsacral with modified oblique methods, respectively. Moreover, 9/40 (22.5%) of the transsacral A/P measurements were <7.3 mm, while all of the modified oblique A/P measurements were >7.3 mm. The mean superior/inferior (S/I) measurements were 18.0 mm for transsacral and 26.2 mm for modified oblique placement. Out of 40 transsacral S/I measurements, 4 (10%) were <14.6 mm, while all the modified oblique S/I measurements were >14.6 mm. In the second part of this study, 10 uninjured cadaveric pelves had unilateral percutaneous IS placed under fluoroscopic guidance (inlet, outlet, and lateral projections) by one orthopedic traumatologist. The final position of all 10 screws was confirmed on fluoroscopy by two independent orthopedic trauma surgeons. The first 5 screws were placed by using transsacral pelvic landmarks. Modified landmarks guided the other 5 screws. The accuracy of final screw position was determined by "postoperative" CT scans interpreted by a blinded musculoskeletal radiologist. The screws inserted using transsacral pelvic landmarks were errant in 3 of the 5 cases. Neurovascular complications could be expected from the extraosseous position of all 3 screws. All 5 screws were located within the confines of the S-1 segment by means of the modified oblique technique. Thus, the modified oblique placement technique allowed greater accuracy and reliability over transsacral landmarks in placing percutaneous ISs. The use of the modified oblique pelvic landmarks is warranted during percutaneous iliosacral screw stabilization of the posterior pelvis. PMID- 11011777 TI - Evaluation and treatment of high subtrochanteric femur fractures. AB - This retrospective study reviews the biomechanical factors and surgical approaches for the treatment of high subtrochanteric femur fractures to determine whether management should be directed toward using an intramedullary or extramedullary device. Results have demonstrated that both plates and intramedullary implants work equally well, with intramedullary implants resulting in significant decreases in surgical times and blood losses. Because intramedullary devices can now be placed through percutaneous trochanteric insertions, they have become more attractive for the management of these injuries. However, intramedullary devices may be difficult to use in fractures presenting with a trochanteric extension, and adjunctive reduction techniques are still required to obtain fracture reductions and prevent varus malunions. When deciding on which technique to use, surgeons should evaluate the fracture pattern and determine whether the implants and techniques that are familiar to them will allow good functional outcomes, high rates of unions, and low rates of complications. PMID- 11011776 TI - Free-flap reconstruction of traumatic lower extremity wounds. AB - A retrospective review of 20 free-tissue transfers for reconstruction of traumatic lower extremity wounds was undertaken. The wounds consisted of Gustilo type IIIb injuries resulting from motor vehicle accidents, falls, motorcycle and four-wheel, all-terrain vehicle accidents, and a laceration from a lawnmower blade. Fifty percent of the procedures were for open tibia fractures. The most common flap used was the rectus abdominis, followed by the latissimus dorsi, serratus anterior, and radial forearm. There were two major complications resulting in flap loss. Adequate and multiple debridements, selection of the appropriate flap, meticulous surgical technique, and postoperative care continue to be the essentials of free-flap coverage. With a motivated patient, limb salvage and return to function can be achieved after most high-energy injuries. PMID- 11011778 TI - Femoral recon nails for metastatic disease: indications, technique, and results. AB - The senior author has used 41 femoral recon nails of one design in 41 consecutive patients with 30 impending and 11 pathologic femur fractures. All nails were inserted with minimally invasive surgical technique. The hospital stay was shorter (average, 5.6 days) and discharge to home was more likely (77%) after stabilization of impending pathologic fractures compared with fixation of completed fractures (length of hospital stay averaged 7.8 days, with a 36% discharge-to-home rate), as patients with completed fractures required a higher level of postoperative care. Fixation prior to fracture occurrence results in predictably better early results than that of fixation after fracture occurrence, confirming the benefit of prophylactic fixation of impending pathologic fractures. To date, there have been no fixation failures and only one significant complication: a nonfatal pulmonary embolus. This technique provides a safe and effective method of stabilizing properly selected femoral lesions resulting from metastatic disease, especially impending pathologic fractures. PMID- 11011779 TI - Impact of enteral and parenteral nutrition on hepatic and muscle glucose metabolism. AB - Liver and muscle metabolism were assessed in dogs adapted to long-term total parenteral (TPN) and enteral (TEN) nutrition. Studies were done in 13 conscious long-term catheterized dogs in which sampling (artery, portal and hepatic vein, and iliac vein), infusion catheters (inferior vena cava, duodenum), and transonic flow probes (hepatic artery, portal vein, and iliac artery) were implanted. Fourteen days after surgery dogs were grouped to receive TPN or TEN. After 5 days of TPN/TEN, substrate balances across the liver and limb were assessed. The liver was a marked net consumer of glucose in both groups (23.6 +/- 3.3 vs 22.6 +/- 2.8 micromol x kg(-1) x min(-1), TPN vs TEN) despite near normoglycemia (6.5 +/- 0.3 vs 6.7 +/- 0.2 mmol/L). Arterial insulin levels were higher during TEN (96 +/- 6 vs 144 +/- 30 pmol/L; p < .05). The majority (79 +/- 13 vs 76% +/- 7%) of the glucose taken up by the liver was released as lactate. Despite higher insulin levels during TEN the nonsplanchnic tissues consumed a lessor quantity of glucose (25.9 +/- 3.3 vs 16.1 +/- 3.9 micro x mol x kg(-1) x min(-1)). In summary, the liver undergoes a profound adaptation to TPN and TEN making it a major site of glucose uptake and conversion to lactate irrespective of the route of nutrient delivery. However, the insulin requirements are higher with TEN possibly secondary to impaired peripheral glucose removal. PMID- 11011780 TI - Individual neuropeptides regulate gut-associated lymphoid tissue integrity, intestinal immunoglobulin A levels, and respiratory antibacterial immunity. AB - BACKGROUND: Total parenteral nutrition (TPN) leads to atrophy of the gut associated lymphoid tissue (GALT) and a significant decrease in intestinal immunoglobulin A (IgA) levels, a major constituent of mucosal immunity. Bombesin (BBS) prevents TPN-induced GALT atrophy and maintains intestinal IgA levels. BBS, a neuropeptide analogous to gastrin-releasing peptide in humans, stimulates the release of other gut neuropeptides including cholecystokinin (CCK), gastrin, and neurotensin (NT). This study investigates the ability of CCK, gastrin, or NT to individually prevent TPN-induced GALT atrophy and preserve respiratory immunity. METHODS: Experiment 1: Male mice were randomly assigned to receive chow, TPN, TPN plus CCK, TPN plus gastrin, or TPN plus NT. After 5 days of feeding, Peyer's patches (PP) from the proximal and distal small bowel were harvested and analyzed for cell yields. PP cells were also analyzed for GALT cell type. Small bowel IgA levels were measured by enzyme-linked immunosorbent assay (ELISA). Experiment 2: Mice were randomly assigned to receive either liposomes containing Pseudomonas antigen or liposomes without antigen. After 10 days, mice were randomly assigned to the same five treatment groups, fed for 5 days, and then given intratracheal Pseudomonas. Mortality was assessed after 48 hours. RESULTS: Experiment 1: GALT cell reductions due to IV-TPN were greater in the distal than proximal small bowel. All three neuropeptides prevented most TPN-induced GALT atrophy due mainly to the maintenance of the B-cell and T-cell populations in the PP of the distal bowel. Intestinal IgA levels were significantly higher in the animals treated with neuropeptides than animals treated with TPN only; however, these IgA levels were not maintained at levels observed in chow-fed animals. Experiment 2: Immunization resulted in significantly lower mortality in animals fed chow, TPN plus CCK, and TPN plus gastrin. TPN alone and TPN plus NT resulted in loss of immunity and mortality rate at comparable levels to unimmunized animals. CONCLUSIONS: Supplementation of IV-TPN with CCK, gastrin, and NT prevents GALT atrophy, primarily in the distal bowel. Intestinal IgA levels improve but not to normal levels. CCK and gastrin reversed IV-TPN-induced effects on antibacterial pneumonia in immunized animals while NT did not. PMID- 11011782 TI - Malnutrition impairs CD11b/CD18 expression on circulating polymorphonuclear neutrophils and subsequent exudation into inflammatory sites in the early phase of glycogen-induced murine peritonitis. AB - BACKGROUND: The effects of malnutrition on polymorphonuclear neutrophil (PMN) exudation are not well understood. The purpose of this study was to examine the effects of short-term dietary restriction on adhesion molecule expression on circulating PMNs and PMN exudation into the inflamed site in a glycogen-induced peritonitis model. METHODS: Twelve mice were randomly assigned to one of two groups. The ad libitum and diet-restricted groups received mouse chow ad libitum (estimated consumption: 132 g/kg per day) and 33 g/kg per day, respectively, for 7 days. Then, 2 mL of a 1% glycogen solution was intraperitoneally administered to all mice. After 4 hours, the animals were killed. Whole blood was drawn by cardiac puncture. Peritoneal exudative cells were harvested by lavaging the peritoneal cavity. Expressions of CD11b, CD18, and CD62L were measured by flow cytometry. RESULTS: Dietary restriction did not affect the numbers of circulating leukocytes, PMNs, or monocytes. However, CD11b and CD18 expressions on circulating PMNs were significantly lower in the diet-restricted than in the ad libitum group. In contrast, CD62L expression on circulating PMNs was not affected by dietary restriction. The number of exudative PMNs was significantly lower in the diet-restricted group than in the ad libitum group. The expressions of CD11b, CD18 and CD62L on exudative PMNs were unaffected by dietary restriction. There was a significant positive correlation between exudative PMN numbers and CD18 expression on circulating PMNs. CONCLUSIONS: Severe dietary restriction in our murine model decreased beta2 integrin expression on circulating PMNs and inhibited PMN exudation into inflamed sites in the early phase of inflammation. These events may increase susceptibility to bacterial infection. Nutritional replenishment may improve host defense in part by enhancing PMN adhesion molecule expression. PMID- 11011781 TI - Glutamine-enriched total parenteral nutrition maintains intestinal interleukin-4 and mucosal immunoglobulin A levels. AB - BACKGROUND: Total parenteral nutrition (TPN) prevents progressive malnutrition but fails to maintain intestinal gut-associated lymphoid tissue (GALT) or established respiratory antiviral or antibacterial mucosal immunity. Our previous work demonstrated that decreases in intestinal immunoglobulin A (IgA) were associated with decreases in Th2-type IgA-stimulating cytokines, interleukin (IL) 4 and IL-10. Because glutamine supplementation of TPN partially preserves respiratory defenses and normalizes GALT, we investigated the ability of parenteral glutamine to normalize respiratory and intestinal IgA levels and measured Th2 cytokines in intestinal homogenates. METHODS: Animals were cannulated and randomly assigned to receive chow (n = 17), TPN (n = 18), or an isonitrogenous, isocaloric TPN solution formulated by removing the appropriate amount of amino acids and replacing them with 2% glutamine (n = 18) for 5 days. Respiratory tract and intestinal washings were obtained for IgA and the intestine homogenized and analyzed for IL-4 and IL-10. RESULTS: TPN decreased intestinal and respiratory IgA in association with decreases in intestinal IL-4 and IL-10 compared with chow-fed animals. Glutamine significantly improved respiratory and intestinal IgA levels, significantly improved IL-4 compared with TPN animals, and maintained IL-10 levels midway between chow-fed and TPN animals. CONCLUSIONS: Glutamine-enriched TPN preserved both extraintestinal and intestinal IgA levels and had a normalizing effect on Th2-type IgA-stimulating cytokines. PMID- 11011783 TI - Arginine supplementation is well tolerated but does not enhance mitogen-induced lymphocyte proliferation in elderly nursing home residents with pressure ulcers. AB - BACKGROUND: Immune function declines with age, increasing risk for infection and delaying wound healing. Arginine enhances immune function and healing of standardized wounds in healthy elderly persons. The purpose of this study was to determine what level of arginine supplementation was orally and metabolically tolerated and effective in enhancing immune function in elderly persons with pressure ulcers. METHODS: Residents with one or more pressure ulcers were recruited from two local nursing homes. Subjects were randomized to receive 0 g (n = 10; age, 82 +/- 3 years), 8.5 g (n = 11; 81 +/- 3 years), or 17 g (n = 11; 87 +/- 2 years) of supplemental arginine each day for 4 weeks. Oral tolerance, ie, absence of nausea, vomiting, abdominal distention, or diarrhea, was assessed daily. Metabolic tolerance was assessed weekly by evaluating serum electrolytes. Lymphocyte proliferation to phytohemagglutinin and interleukin 2 production were measured at baseline and after 4 weeks of supplementation as indicators of immune function. RESULTS: Supplemental arginine significantly increased plasma arginine levels and was orally and metabolically tolerated with no complaints of abdominal distress or no clinically relevant changes in electrolyte levels among groups. Lymphocyte proliferation and interleukin 2 production were significantly different between nursing homes. When data from nursing homes were considered individually, arginine supplementation did not enhance the proliferative response. In subjects from nursing home 2 only, there was a 38% and 75% decrease (p < .05) in lymphocyte proliferation with 8.5 and 17 g of supplemental arginine, respectively. Interleukin 2 production was no different among supplementation groups. CONCLUSIONS: Pharmacologic doses of arginine were well tolerated but did not enhance lymphocyte proliferation or interleukin 2 production in nursing home residents with pressure ulcers. CLINICAL RELEVANCY: Enteral formulas supplemented with pharmacologic levels of arginine are frequently administered to elderly persons. This study demonstrates that the very old can tolerate these nitrogen loads if baseline renal function is normal and fluid intake is encouraged. Further research needs to be completed investigating the effect of arginine supplementation on immune function in this population before recommending arginine use. PMID- 11011784 TI - Enterotrophic effect of insulin-like growth factor-I but not growth hormone and localized expression of insulin-like growth factor-I, insulin-like growth factor binding protein-3 and -5 mRNAs in jejunum of parenterally fed rats. AB - BACKGROUND: Administration of insulin-like growth factor (IGF)-I, but not growth hormone (GH), stimulates mucosal hyperplasia in surgically stressed rats with intestinal atrophy induced by hypocaloric total parenteral nutrition (TPN). Our aim was to characterize the basis for this disparity in enterotrophic action by assessing the relationships between stimulation of intestinal growth, nutritional adequacy, and localization of expression of IGF-I, insulin-like growth factor binding protein (IGFBP)-3 and IGFBP-5 mRNAs in jejunum. METHODS: Rats were maintained with TPN for 8 days and treated with IGF-I or GH and adequate nutrition for 5 days after recovery from surgery. Jejunal mass, morphology, and sucrase activity were assessed. Localization of expression of IGF-I, IGFBP-3, and IGFBP-5 mRNAs in jejunum was accomplished by in situ hybridization. RESULTS: Serum IGF-I and body weight gain were significantly increased by IGF-I or GH. Jejunal mucosal dry mass, morphology, and sucrase activity were improved with IGF I but not GH. There were no differences in IGF-I mRNA. IGFBP-3 mRNA was localized in the lamina propria of the villi. IGF-I or GH stimulated IGFBP-3 expression. IGF-I strongly stimulated IGFBP-5 expression in the lamina propria and the muscularis and induced a twofold increase in IGFBP-5 mRNA based on RNase protection assay of intact jejunum total RNA. GH induced a modest increase in IGFBP-5 expression in the muscularis with no effect on intact jejunum total RNA. CONCLUSIONS: The GH resistance observed in the jejunal mucosa of TPN rats cannot be fully explained by inadequate nutrition. The expression of IGFBP-5 in the lamina propria suggests it may modulate the enterotrophic action of exogeneous IGF-I. PMID- 11011785 TI - Hazard analysis and critical control point system approach in the evaluation of environmental and procedural sources of contamination of enteral feedings in three hospitals. AB - BACKGROUND: The administration of contaminated diets may contribute to severe infections, mainly in immunosuppressed patients. To determine the microbiologic quality of enteral feedings and the critical control points involved in the processing of the formulas, a study was carried out in three hospitals in Sao Paulo, Brazil. METHODS: One hundred sixteen diets were evaluated according to the system known as hazard analysis and critical control points (HACCP). The techniques included the monitoring of the cleaning and disinfection of utensils, surfaces, and equipment; time and temperature controls; and microbiologic analyses that comprised the enumeration of facultative aerobic mesophilic bacteria, yeasts and molds, coliforms, and Escherichia coli. The diets were assessed based on the British Dietetic Association and the Food and Drug Administration standards. RESULTS: The hospital-formulated diets and the commercially made powdered feeds presented statistically significant higher counts of mesophilic bacteria and percentages of no compliance with the standards for coliforms when compared with the commercially prepared, ready-to-feed enteral formula supplied in cans. According to the British standards, 77% and 38% of the diets were inadequate for mesophilic and coliform bacteria, respectively, just after preparation. After 24-hour storage in refrigerators, the percentages rose to 83% and 45%, respectively; the mesophilic and coliforms counts were also significantly higher. The following critical control points (CCP) were identified: cleaning and disinfecting of surfaces, utensils, and equipment; the time spent in preparation; the water used for reconstitution; the final temperature of the diet; the exposure to room temperatures; the lack of a chilling step; the refrigeration temperature; the inadequate disinfecting of the handlers' hands; and the lack of external cleaning and disinfecting of the cans before opening. CONCLUSIONS: The results of this study are worrisome and show the need for implementation of good practices of hygiene and handling of the diets, and the regular monitoring of their preparation. The microbiologic quality of the enteral diets was compromised according to international standards, representing a potential risk of infection to the patients who require enteral nutrition. PMID- 11011786 TI - A multicenter, prospective study of the placement of transpyloric feeding tubes with assistance of a magnetic device. The Magnet-Guided Enteral Feeding Tube Study Group. AB - BACKGROUND: Placement of feeding tubes in the transpyloric position can be helpful in the management of enterally fed patients with pancreatitis, gastric atony, enterocutaneous fistulae, or pulmonary aspiration risk. The attainment of transpyloric position is difficult, and numerous techniques have been proposed to help in achieving this location. Recently, the use of a magnet-tipped feeding tube, dragged into proper position with an external magnet, has been described with an excellent success rate. METHODS: At 10 participating institutions, practitioners were trained in the use of the device. Successful tube placement was determined by abdominal radiograph. RESULTS: One hundred fifty-six tube placements were attempted. Transpyloric position was obtained in 60%. Placement into the third portion of the duodenum or distal was obtained in only 32%. Analysis of the data did not reveal a learning curve at the institutions, and 7 of 10 had a 50% or lower success rate. CONCLUSIONS: Placement of feeding tubes with the assistance of a magnetic device was infrequently successful at the majority of institutions where it was attempted. We report a lower success rate than the original article, which described an 88% success rate of transpyloric intubation. Although this technique has a high failure rate, some individuals seem to be very successful using it, which could reduce the need for endoscopy or transport for the placement of feeding tubes. PMID- 11011787 TI - A novel efficient method to identify beta-glucuronidase activity in rat small intestine. AB - BACKGROUND: Recent epidemiologic studies promote the notion that high intake of food rich in phytochemicals protects against degenerative diseases such as coronary heart diseases and cancer. Phytochemicals are detoxified in mammalian tissues by conjugation with glucuronic acid yielding less active glucuronide conjugates. However, in several tissues glucuronide conjugates are reactivated by the cleaving enzyme beta-glucuronidase. The aim of the present study was to develop a routinely manageable, rapid technique to localize the beta glucuronidase activity in the small intestinal tissue. METHODS: Histologic slices of rat duodenum, jejunum, and ileum were incubated with a specific chromogenic beta-glucuronidase substrate, 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X GlcU). After enzymatic cleavage, X-GlcU yields 5-bromo-4-chloro-3-indol, a dark blue crystalline precipitate easily monitored by light microscopic technique. RESULTS: The number and intensity of the crystals were highest in the jejunum and lowest in the ileum. In all three sections of the small intestine, the highest activity was observed at the villar tip and in the tela submucosa and only moderate activity in other layers of the intestinal tissue. CONCLUSIONS: By using the X-GlcU-technique, it could be demonstrated convincingly that beta glucuronidase exists in all three segments of the rat small intestine. The proposed method is an efficient, simple, and convenient method to visualize beta glucuronidase activity. PMID- 11011788 TI - Digital VLSI algorithms and architectures for support vector machines. AB - In this paper, we propose some very simple algorithms and architectures for a digital VLSI implementation of Support Vector Machines. We discuss the main aspects concerning the realization of the learning phase of SVMs, with special attention on the effects of fixed-point math for computing and storing the parameters of the network. Some experiments on two classification problems are described that show the efficiency of the proposed methods in reaching optimal solutions with reasonable hardware requirements. PMID- 11011789 TI - Implementation of a pulse coupled neural network in FPGA. AB - The Pulse Coupled neural network, PCNN, is a biologically inspired neural net and it can be used in various image analysis applications, e.g. time-critical applications in the field of image pre-processing like segmentation, filtering, etc. a VHDL implementation of the PCNN targeting FPGA was undertaken and the results presented here. The implementation contains many interesting features. By pipelining the PCNN structure a very high throughput of 55 million neuron iterations per second could be achieved. By making the coefficients re configurable during operation, a complete recognition system could be implemented on one, or maybe two, chip(s). Reconsidering the ranges and resolutions of the constants may save a lot of hardware, since the higher resolution requires larger multipliers, adders, memories etc. PMID- 11011791 TI - VLSI implementation of neural networks. AB - Currently, fuzzy controllers are the most popular choice for hardware implementation of complex control surfaces because they are easy to design. Neural controllers are more complex and hard to train, but provide an outstanding control surface with much less error than that of a fuzzy controller. There are also some problems that have to be solved before the networks can be implemented on VLSI chips. First, an approximation function needs to be developed because CMOS neural networks have an activation function different than any function used in neural network software. Next, this function has to be used to train the network. Finally, the last problem for VLSI designers is the quantization effect caused by discrete values of the channel length (L) and width (W) of MOS transistor geometries. Two neural networks were designed in 1.5 microm technology. Using adequate approximation functions solved the problem of activation function. With this approach, trained networks were characterized by very small errors. Unfortunately, when the weights were quantized, errors were increased by an order of magnitude. However, even though the errors were enlarged, the results obtained from neural network hardware implementations were superior to the results obtained with fuzzy system approach. PMID- 11011790 TI - A programmable VLSI filter architecture for application in real-time vision processing systems. AB - An architecture is proposed for the realization of real-time edge-extraction filtering operation in an Address-Event-Representation (AER) vision system. Furthermore, the approach is valid for any 2D filtering operation as long as the convolutional kernel F(p,q) is decomposable into an x-axis and a y-axis component, i.e. F(p,q)=H(p)V(q), for some rotated coordinate system [p,q]. If it is possible to find a coordinate system [p,q], rotated with respect to the absolute coordinate system a certain angle, for which the above decomposition is possible, then the proposed architecture is able to perform the filtering operation for any angle we would like the kernel to be rotated. This is achieved by taking advantage of the AER and manipulating the addresses in real time. The proposed architecture, however, requires one approximation: the product operation between the horizontal component H(p) and vertical component V(q) should be able to be approximated by a signed minimum operation without significant performance degradation. It is shown that for edge-extraction applications this filter does not produce performance degradation. The proposed architecture is intended to be used in a complete vision system known as the Boundary-Contour-System and Feature Contour-System Vision Model, proposed by Grossberg and collaborators. The present paper proposes the architecture, provides a circuit implementation using MOS transistors operated in weak inversion, and shows behavioral simulation results at the system level operation and electrical simulation and experimental results at the circuit level operation of some critical subcircuits. PMID- 11011792 TI - Convergence analysis of cascade error projection--an efficient learning algorithm for hardware implementation. AB - In this paper, we present a mathematical foundation, including a convergence analysis, for cascading architecture neural network. Our analysis also shows that the convergence of the cascade architecture neural network is assured because it satisfies Liapunov criteria, in an added hidden unit domain rather than in the time domain. From this analysis, a mathematical foundation for the cascade correlation learning algorithm can be found. Furthermore, it becomes apparent that the cascade correlation scheme is a special case from mathematical analysis in which an efficient hardware learning algorithm called Cascade Error Projection(CEP) is proposed. The CEP provides efficient learning in hardware and it is faster to train, because part of the weights are deterministically obtained, and the learning of the remaining weights from the inputs to the hidden unit is performed as a single-layer perceptron learning with previously determined weights kept frozen. In addition, one can start out with zero weight values (rather than random finite weight values) when the learning of each layer is commenced. Further, unlike cascade correlation algorithm (where a pool of candidate hidden units is added), only a single hidden unit is added at a time. Therefore, the simplicity in hardware implementation is also achieved. Finally, 5 to 8-bit parity and chaotic time series prediction problems are investigated; the simulation results demonstrate that 4-bit or more weight quantization is sufficient for learning neural network using CEP. In addition, it is demonstrated that this technique is able to compensate for less bit weight resolution by incorporating additional hidden units. However, generation result may suffer somewhat with lower bit weight quantization. PMID- 11011793 TI - Simulink-based HW/SW codesign of embedded neuro-fuzzy systems. AB - We propose a semi-automatic HW/SW codesign flow for low-power and low-cost Neuro Fuzzy embedded systems. Applications range from fast prototyping of embedded systems to high-speed simulation of Simulink models and rapid design of Neuro Fuzzy devices. The proposed codesign flow works with different technologies and architectures (namely, software, digital and analog). We have used The Mathworks' Simulink environment for functional specification and for analysis of performance criteria such as timing (latency and throughput), power dissipation, size and cost. The proposed flow can exploit trade-offs between SW and HW as well as between digital and analog implementations, and it can generate, respectively, the C, VHDL and SKILL codes of the selected architectures. PMID- 11011794 TI - Automating parallel implementation of neural learning algorithms. AB - Neural learning algorithms generally involve a number of identical processing units, which are fully or partially connected, and involve an update function, such as a ramp, a sigmoid or a Gaussian function for instance. Some variations also exist, where units can be heterogeneous, or where an alternative update technique is employed, such as a pulse stream generator. Associated with connections are numerical values that must be adjusted using a learning rule, and and dictated by parameters that are learning rule specific, such as momentum, a learning rate, a temperature, amongst others. Usually, neural learning algorithms involve local updates, and a global interaction between units is often discouraged, except in instances where units are fully connected, or involve synchronous updates. In all of these instances, concurrency within a neural algorithm cannot be fully exploited without a suitable implementation strategy. A design scheme is described for translating a neural learning algorithm from inception to implementation on a parallel machine using PVM or MPI libraries, or onto programmable logic such as FPGAs. A designer must first describe the algorithm using a specialised Neural Language, from which a Petri net (PN) model is constructed automatically for verification, and building a performance model. The PN model can be used to study issues such as synchronisation points, resource sharing and concurrency within a learning rule. Specialised constructs are provided to enable a designer to express various aspects of a learning rule, such as the number and connectivity of neural nodes, the interconnection strategies, and information flows required by the learning algorithm. A scheduling and mapping strategy is then used to translate this PN model onto a multiprocessor template. We demonstrate our technique using a Kohonen and backpropagation learning rules, implemented on a loosely coupled workstation cluster, and a dedicated parallel machine, with PVM libraries. PMID- 11011795 TI - FPNA: interaction between FPGA and neural computation. AB - Neural networks are usually considered as naturally parallel computing models. But the number of operators and the complex connection graph of standard neural models can not be directly handled by digital hardware devices. More particularly, several works show that programmable digital hardware is a real opportunity for flexible hardware implementations of neural networks. And yet many area and topology problems arise when standard neural models are implemented onto programmable circuits such as FPGAs, so that the fast FPGA technology improvements can not be fully exploited. Therefore neural network hardware implementations need to reconcile simple hardware topologies with complex neural architectures. The theoretical and practical framework developed, allows this combination thanks to some principles of configurable hardware that are applied to neural computation: Field Programmable Neural Arrays (FPNA) lead to powerful neural architectures that are easy to map onto FPGAs, thanks to a simplified topology and an original data exchange scheme. This paper shows how FPGAs have led to the definition of the FPNA computation paradigm. Then it shows how FPNAs contribute to current and future FPGA-based neural implementations by solving the general problems that are raised by the implementation of complex neural networks onto FPGAs. PMID- 11011796 TI - Effects of hyperbaric oxygen on ventricular performance, pulmonary blood volume, and systemic and pulmonary vascular resistance. AB - The cardiovascular effects of hyperbaric (3 atm abs) air, 100% oxygen, and hyperbaric oxygen (HBO2) at 3 atm abs were investigated in 13 pentobarbital anesthetized dogs. We measured heart rate, arterial pressure, pulmonary artery pressure, right atrial pressure, left and right ventricular pressure, and cardiac output. From these we determined end diastolic pressure, +/-maximal dp/dt, maximal (dp/p), end systolic elastance, cardiac work, and systemic (SVR) and pulmonary vascular resistance (PVR). Pulmonary blood volume was obtained from the mean transit time of ascorbic acid. The significant results with HBO2 were a decrease in heart rate, cardiac output, and cardiac work. All left ventricular performance indices decreased, without a change in preload or afterload. In contrast, only right ventricular -dp/dt decreased. SVR increased but PVR did not change; 100% O2 produced similar but less pronounced responses. Hyperbaric air had only mild effects. Pulmonary blood volume and lung wet/dry ratio did not change. Our data suggest that HBO2 may act by a differential effect on the autonomic innervation of the right and left ventricles. The resultant ventricular imbalance may be of clinical importance in the mechanism of pulmonary edema in patients in congestive heart failure undergoing hyperbaric therapy. PMID- 11011797 TI - Influence of gender and menstrual cycle on a cold air tolerance test and its relationship to thermosensitivity. AB - This investigation evaluated the influence of gender and phase of menstrual cycle [follicular (FOL): Days 2-6) and luteal (LUT: Days 19-24)] on a cold air tolerance test (CATT: 90-min of exposure to 5 degrees C air) in 8 females (22.7 +/- 3.0 yr) and 15 males (22.3 +/- 2.9 yr). In addition, central thermosensitivity (beta; W x kg(-1) x degrees C(-1) [i.e., the slope of the relationship between the decrease in esophageal temperature (Tes) and the increase in heat production (HP)], gathered during a separate water trial in 20 degrees C water, was correlated to the change (delta) in Tes and HP across the 90 min of resting exposure during the CATT. Analysis of variance revealed no significant differences between phase of menstrual cycle or gender for HP, mean skin temperature (Tsk), and insulation; however, a main effect for time for these parameters was demonstrated. Despite these similarities, Tes differed (P < 0.05) between males and females. Additionally, no relationship was found between beta and deltaHP and deltaTes in the males and females. Also, there was no relationship between beta and thermoregulation during the CATT in these subjects. These data suggest that menstrual cycle phase did not cause a differential response in Tes, Tsk, and HP during a CATT. Furthermore, women maintained a higher Tes than men during the CATT despite similarities in HP and Tsk. PMID- 11011798 TI - Effects of smoking on cost and duration of hyperbaric oxygen therapy for diabetic patients with non-healing wounds. AB - During this study to determine the effects of smoking on diabetic patients undergoing hyperbaric oxygen therapy (HBO2T) for nonhealing wounds, one physician visited five hyperbaric facilities and reviewed records on 1,006 patients who had received HBO2T for diabetic wounds. Smoking history was documented on 469 patients, while 180 patients had complete information on number of HBO2Ts, outcome, age, duration of diabetes, transcutaneous oxygen baseline in air at ambient conditions, Wagner score of the worst wound, smoking history, and intensity of treatment. These factors were statistically significant predictors of treatment outcome using multiple regression modeling. No difference was found between smokers with less than 10 pack-years of cumulative history and nonsmokers. After that point there was a significant increase in the number of HBO2Ts needed to produce at least some healing in smokers vs. patients who had never smoked. The average patient with a greater than 10 pack-year smoking history who benefited from treatment was estimated to need between 8 and 14 more HBO2Ts. This translates into an added treatment cost of $4,000 to $7,000 for the average patient who has smoked, and an estimated $22-37 million annually for the United States. PMID- 11011799 TI - Hearing function in a hyperbaric environment. AB - Navy divers' hearing function was assessed as part of three saturation deep dives to 1,000 feet of sea water (fsw) to determine explanations for threshold shifts observed under hyperbaric conditions. Across the three deep dives, different aspects of the ear were evaluated, including air- and bone-conduction pure-tone thresholds, real ear probe microphone measurements, auditory evoked potentials, and central auditory processing assessments. Attempts to measure middle ear function and cochlear function (through otoacoustic emissions) were unsuccessful. Baseline measurements were obtained at 0 fsw in air before and after the saturation deep dives. Results showed that some aspects of hearing function remained unchanged with increases in depth. In general, audiometric thresholds at depth were similar to those measured on the surface at 500, 1,000, 2,000, 3,000, and 4,000 Hz. However, hearing sensitivity actually improved at depth at 6,000 and 8,000 Hz. The use of a specially designed sound booth for a pressurized heliox environment yielded significantly lower ambient noise levels and improved the accuracy of threshold measurement. Auditory evoked potential measurements and central auditory processing function were relatively unaffected by changes in depth. Significant changes at depth were seen in ear canal resonance which shifted up in frequency; this finding was attributed to the effect of helium on the hearing mechanism. Because objective measurement of middle ear and inner ear function were not methodologically possible, questions still remain regarding the interpretation of middle and inner ear function at depth. Nonetheless, our overall findings suggest that most aspects of hearing functioning are similar under high atmospheric pressures and in heliox as they are on the surface, with the exception of shifts in ear canal resonance and improvements in audiometric thresholds at high frequencies. PMID- 11011800 TI - Performance of the Baxter Flo-Gard 6201 volumetric infusion pump for monoplace chamber applications. AB - For non-hyperbaric purposes, the Baxter Flo-Gard 6201 volumetric pump is capable of infusing multiple types of fluids at rates of 1-1,999 ml x h(-1). We designed a study to determine flow accuracy of this pump at variable rates, fluid viscosities, and volumes over a range of chamber pressures. For hyperbaric use, the pump pressure sensor was adjusted. Sodium chloride solution 0.9% (NS), enteral formula, and packed red blood cells (PRBC) were infused at varying rates from 86.1 to 304 kPa (0.85 to 3.0 atm abs). For NS, measured compared to set flow rates ranged from 12.5% to -7.5% at settings of 1 and 5 ml x h(-1) from 86.1 to 304 kPa (0.85 to 3.0 atm abs) pressures, respectively. For NS infusions at a set rate of 100 ml x h(-1), the measured flow was identical to the set rate at all pressures. At flow settings of 1,999 ml x h(-1), the measured flow varied from the set flow by +/-4.9% Enteral infusion at 100 ml x h(-1) showed approximately a 3% increase in the measured vs. set flow rate. PRBC measured flow rates ranged from -0.4 to 6% of the set rate. During chamber compression and decompression, with set flow rates from 1 to 10 ml x h(-1), the measured flow was considerably less than expected during compression and more than expected during decompression. In conclusion, the Baxter Flo-Gard 6201 infusion pump demonstrated acceptable performance for infusing saline, enteral formula, and PRBC at low and high infusion rates into the pressurized monoplace hyperbaric chamber up to 304 kPa (3 atm abs), with the exception of low rates during compression and decompression. PMID- 11011801 TI - Heparins in the new millennium: will unfractionated heparin survive? Introduction. PMID- 11011802 TI - An update on heparins at the beginning of the new millennium. AB - Unfractionated heparin has enjoyed the sole anticoagulant status for almost half a century. Besides an effective anticoagulant, this drug has been used in several additional indications. Despite the development of newer anticoagulant drugs, unfractionated heparin has remained the drug of choice for surgical anticoagulation and interventional cardiology. In the area of hematology and transfusion medicine, unfractionated heparin has continued to play a major role as an anticoagulant drug. The development of low-molecular-weight heparins (LMWHs) represents a refinement for the use of heparin. These drugs represent a class of depolymerized heparin derivatives with a distinct pharmacologic profile that is largely determined by their composition. These drugs produce their major effects by combining with antithrombin and exerting antithrombin and anti-Xa inhibition. In addition, the LMWHs also increase non-antithrombin-dependent effects such as TFPI release, modulation of adhesion molecules, and release of profibrinolytic and antithrombotic mediators from the blood vessels. The cumulative effects of each of the different LMWHs differ and each product exhibits a distinct profile. Initially these agents were developed for the prophylaxis of postsurgical deep-vein thrombosis. However, at this time these drugs are used not only for prophylaxis, but also for the treatment of thrombotic disorders of both the venous and arterial type. To a large extent, the LMWHs have replaced unfractionated heparin in most subcutaneous indications. With the use of these refined heparins, outpatient anticoagulant management has gone through a dramatic evolution. For the first time, patients with thrombotic disorders can be treated in an outpatient setting. Thus, the introduction of LMWHs represents a major advance in improving the use of heparin. The development of the oral formulation of heparin and LMWHs also provides an important area that may impact on the use of heparin and LMWHs. The increased awareness of heparin-induced thrombocytopenia has necessitated the development of newer methods to identify patients at risk of developing this catastrophic syndrome. Furthermore, a strong interest has developed in alternate drugs or the management of patients with this syndrome. Despite the development of alternate anticoagulants that are mostly antithrombin derived (hirudins, hirulog), these agents have failed to provide similar clinical outcome as heparin in many indications. However, antithrombin drugs are useful in the anticoagulant management of heparin-compromised patients. The FDA has approved a recombinant hirudin (Refludan) and a synthetic antithrombin agent, argatroban (Novastan), for this indication. The development of synthetic heparin pentasaccharide and anti-Xa agents may have an impact on the prophylaxis of thrombotic disorders. However, these monotherapeutic agents do not mimic the polytherapeutic actions of heparin. Furthermore, these agents do not inhibit thrombin. Heparin and LMWHs are capable of inhibiting not only factor Xa and thrombin, but other serine proteases in the coagulation network. The only way the newer drugs can mimic the actions of heparin is in combination modalities (polytherapeutic approaches). It has been suggested that newer antiplatelet drugs also exhibit anticoagulant actions. While these drugs may exhibit weak effects on thrombin generation, none of the currently available antiplatelet drugs exhibit any degree of antithrombin actions. It is likely that heparins synergize or augment the effects of the new antiplatelet drugs. Currently, combination approaches are used to anticoagulate patients in these studies. The dosage of heparins has been arbitrarily reduced. This may not be an optimal procedure. Additional clinical studies are needed to study these combinations where the alterations of these drugs are compared. Such combinations will require newer monitoring approaches. The development of oral thrombin agents, GP IIb PMID- 11011803 TI - Development of tinzaparin: a heparinase-digested low-molecular-weight heparin. AB - Although heparin and its properties had been discovered in the early 1920s, the different characteristics associated with different molecular-weight fractions of heparin were only recognized in the late 1970s. Tinzaparin is a low-molecular weight heparin (LMWH) produced by heparinase digestion of heparin. Preclinical research on tinzaparin established that there were no differences in the antithrombotic activity compared with heparin. Clinical studies evaluating tinzaparin vs. standard heparin for thromboprophylaxis of deep-vein thrombosis in general and orthopedic surgery found that tinzaparin was as effective as standard heparin. Tinzaparin was also evaluated vs. standard heparin in the treatment of acute proximal vein thrombosis; time-to-event curves suggested that this LMWH could be more effective than standard heparin. PMID- 11011804 TI - Comparative pharmacokinetics of LMWHs. AB - A variety of pharmaceutical preparations of low-molecular-weight heparins (LMWHs) are available. They belong to the same family of compounds-ie, heparin derivatives with a narrow distribution of mean molecular weights (MWs). LMWHs have different methods of preparation, which result in variations in mean MW, distribution of MW, and pharmacokinetic (PK) and pharmacodynamic (PD) profiles. The mean MW of these compounds ranges from 3,600 to 6,500 daltons. The ratio of anti-Xa (aXa) and anti-IIa (aIIa) activities of the different LMWHs ranges from 1.5 to >10. After subcutaneous (SC) injection of a prophylactic or therapeutic dose, the peak values for plasma aXa or aIIa activity may vary twofold to threefold because of differences in bioavailability, plasma clearance (Clplasma), and half-life (t1/2). The injection of equivalent amounts of product, based on aXa and aIIa international units (IU), may result in different areas under the curve for the respective activities. Although tinzaparin has a high aIIa specific activity per milligram (and consequently, a low aXa/aIIa ratio), SC injection of 40 mg of enoxaparin (4,000 aXa IU) results in a higher aXa peak value in patients with total hip replacement than 4,500 aXa IU of tinzaparin. Differences in aIIa and aXa peak activities are more striking when high doses of LMWHs are used. The activated partial thromboplastin time (aPTT) can be significantly prolonged, an effect that is related to aIIa and aXa activity. The volume of distribution of LMWHs is of the same order of magnitude as that of the plasma volume. The mean retention time of aXa activity varies from 5.2 (dalteparin) to approximately 7 h (enoxaparin, nadroparin). Bioavailability of prophylactic doses of LMWHs ranges from 86% (dalteparin) to 98% (enoxaparin, nadroparin). PK parameters appear to be minimally affected by a patient's age. The Clplasma is different for each LMWH: 16 mL/min enoxaparin, 21 mL/min nadroparin, 33 mL/min dalteparin, 19 mL/min reviparin, and 22 mL/min tinzaparin. Accumulation of product has been observed for almost all LMWHs in patients with renal insufficiency. LMWHs are effective and safe for treatment or prophylaxis of venous thromboembolism during pregnancy, because they do not cross the placenta. No data are available regarding the passage of LMWHs into the milk in lactating women. Although LMWHs are also effective in prevention and treatment of thromboembolic disease in children, optimal use of these agents in pediatric patients has not been determined. In summary, the PD and PK of LMWHs have been well documented and have demonstrated that LMWHs have a more predictable response, a greater bioavailability, and a longer aXa t1/2 than unfractionated heparin. However, their distribution of MW affects their physicochemical and biological properties, as well as PK characteristics. The concept of aXa/aIIa ratio (determined in vitro) does not account for the differing PK of aXa and aIIIa activity in circulating blood. PMID- 11011805 TI - Comparative efficacy of different low-molecular-weight heparins (LMWHs) and drug interactions with LMWH: implications for management of vascular disorders. AB - The low-molecular-weight heparins (LMWHs) are more efficacious and safer than unfractionated heparin (UFH) in the prevention and treatment of venous thrombosis and to a certain extent in the treatment of acute ischemic syndromes. Because of their predictable pharmacokinetics and bioavailability after subcutaneous administration, LMWHs can be more convenient for outpatient use than UFH. Differences in the manufacturing process of LMWHs result in significant structural and molecular weight differences; thus, LMWHs have individual biochemical and pharmacological profiles and are not interchangeable on the basis of either mass or anti-Xa activity. Using thromboelastograph (TEG) and platelet aggregometry, this investigation compared the in vitro efficacy among various LMWHs and examined the interactions between LMWHs and platelet glycoprotein (GP) IIb/IIIa antagonists. TEG was used to determine the ability of platelet and fibrin interactions to augment blood clots, an effect measured under conditions of maximal platelet activation during clot formation accelerated by recombinant human tissue factor (TF). The comparative efficacy of LMWHs on different mediator induced clot retraction in human blood was assessed by TEG, which demonstrated the potency of different LMWHs to inhibit various mediator-induced clot formations under shear. Tinzaparin was relatively more effective in inhibiting TF , lipopolysaccharide-, factor (f) Xa-, and thrombin-induced clot formation under shear. Under these conditions, platelets significantly enhance clot strength (eightfold vs. platelet-free fibrin clots). LMWHs appear to have broader efficacy than other anticoagulants. Abciximab and roxifiban further inhibited clot strength by affecting the transmission of platelet contractile force to fibrin by platelet GPIIb/IIIa receptors. Subtherapeutic doses of tinzaparin combined with abciximab or roxifiban resulted in a distinct synergy that improved anticoagulant and antiplatelet efficacy mediated by TF, fXa, or thrombin. As these data suggest, the combination of low-dose tinzaparin with low-dose GPIIb/IIIa antagonists (abciximab, roxifiban) may be efficacious in the prevention and treatment of various thromboembolic disorders. PMID- 11011806 TI - Prophylaxis and treatment of deep-vein thrombosis. AB - In 1980, unfractionated heparin (UFH) was the established agent for the prophylaxis of venous thromboembolic (VTE) disease in patients undergoing general surgery. VTE prophylaxis was the first indication in which low-molecular-weight heparins (LMWHs) were tested. Approximately 40 trials have demonstrated that LMWHs are at least as effective and safe as UFH. LMWHs exhibit a number of advantages over UFH, including ease of administration, convenient once-daily dosing, and facilitation of outpatient management. The ideal time of administration and the dose of the initial one or two injections of LMWH remain unresolved issues. LMWHs are used with increasing frequency in the treatment of acute deep vein thrombosis (DVT), having been studied in comparison to UFH in 16 major clinical trials. LMWHs are at least as effective as UFH in the prevention of VTE, but higher doses than those used for prophylaxis are required. There is still an ongoing debate about whether the daily dose should be administered in one or two subcutaneous injections. In some recent studies, symptomatic new DVTs or pulmonary emboli (PE) were the primary end points, which had to be verified by objective methods, but such end points may be not be sensitive enough to detect major differences in the efficacy of different LMWHs. PMID- 11011807 TI - Comparative efficacy of low-molecular-weight heparins in orthopedic surgery. AB - The low-molecular-weight heparins (LMWHs) have been evaluated in the prevention of postoperative thromboembolic disease and have been found to be clinically efficacious and safe. Studies conducted in similar surgery settings have resulted in significantly different reductions in the incidence of deep-vein thrombosis, making an analysis of grouped studies complex. Only two studies have reported head-to-head comparisons of two different LMWHs and showed no difference in clinical end points between enoxaparin and either reviparin or tinzaparin. Our study at the Aalborg hospital in Denmark, comparing two different dosage regimens of LMWH tinzaparin, supported the conclusions of the head-to-head comparative studies. LMWHs are distinct drug entities that cannot be interchanged at equivalent anti-Xa dosages, and the interpretation of their relative efficacy and safety may be biased by the degree of clinical experience of the individual investigators. PMID- 11011808 TI - An equivalence study of two low-molecular-weight heparins in the prevention and treatment of deep-vein thrombosis after total hip replacement. AB - Low-molecular-weight heparins have been evaluated against unfractionated heparin and have been shown to have clinical benefits in the perioperative setting during orthopedic surgery. Two low-molecular-weight heparins, tinzaparin and enoxaparin, were assessed for equivalency in efficacy and safety in 499 patients undergoing elective hip replacement. The two anticoagulants were found to be effective and safe in this setting in the specified patient type. No statistical difference was observed in the incidence of bleeding. Clinical side effects tended to be greater in patients with a higher body mass index, which could serve as a possible indicator for the risk of deep vein thrombosis. PMID- 11011809 TI - Low-molecular-weight heparin in the treatment of venous thromboembolism. AB - The low-molecular-weight heparins (LMWHs) have been evaluated in the prevention and treatment of deep-vein thrombosis and pulmonary embolism. LMWHs have been found to be safe and effective in this clinical setting and have advantages over unfractionated heparin. These advantages include less serious and less frequent therapeutic complications. The favorable pharmacokinetic profile of LMWHs compared with heparin has allowed for safe, effective, and convenient treatment of patients with venous thromboembolism. Use of LMWHs ultimately results in considerable cost savings for the health care system. PMID- 11011810 TI - Low-molecular-weight heparin and cancer. AB - Heparin is a familiar anticoagulant drug with properties that may impede tumor growth; it modifies properties of cells that contribute to malignant dissemination such as angiogenesis, growth factor and protease activity, immune function, proliferation, and gene expression. Heparin has antitumor effects in animal models of malignancy, and studies in human malignancy show improved cancer outcome with heparin treatment. Meta-analyses comparing unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) for treatment of deep-vein thrombosis have shown apparent substantial improvement in cancer outcome in the subset of patients with malignancy who were randomly assigned to receive LMWH. This experience, together with the favorable pharmacokinetic properties of LMWH, provides a rationale for prospective clinical trials of LMWH in patients with cancer. Such trials should provide (a) definitive data on possible antitumor effects of this treatment, (b) insight into possible heterogeneous responses to heparin treatment among different histological types and stages of malignancy, and (c) a setting for exploring mechanisms of antineoplastic effect in human malignancy. PMID- 11011812 TI - Heparins in the new millennium: will unfractionated heparin survive? Epilogue. PMID- 11011811 TI - Antithrombotics and anticoagulants in coronary syndromes and stroke. AB - The low-molecular-weight heparins (LMWHs) have been proven superior to placebo in reducing the incidence of acute coronary ischemic syndromes. Comparative studies vs. unfractionated heparin have not demonstrated superiority in favor of the LMWH dalteparin. In the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-wave Coronary Events (ESSENCE) trial, enoxaparin was demonstrated to have a benefit over heparin. The results have contributed to a better understanding of the relative efficacy of LMWHs in acute coronary syndromes. A second trial with enoxaparin supported the conclusions of the ESSENCE trial. The antithrombotic effects of LMWHs have also been evaluated for the management of ischemic stroke with varied results. A trial assessing tinzaparin in acute ischemic stroke has completed enrollment, and its results may shed new light on the use of an LMWH for the management of stroke. PMID- 11011813 TI - CHOP Infant Coma Scale ("Infant Face Scale"): a novel coma scale for children less than two years of age. AB - The Glasgow Coma Scale (GCS) is the most frequently used tool worldwide for assessing the severity of neurologic injury after brain trauma, although applying this scale to infants and younger children can be problematic. The CHOP Infant Coma Scale, or Infant Face Scale (IFS), is a novel scale for children under 2 years of age which differs from other pediatric coma scales in the following ways: (1) it relies on objective behavioral observations; (2) it assesses cortical as well as brainstem function; (3) it parallels the GCS in scoring but is based on infant-appropriate behaviors; and (4) it can be applied to intubated patients. We report the results of a prospective study designed to compare interrater reliability between the IFS and GCS in children less than 2 years of age. Seventy-five hospitalized children less than 2 years of age were assessed simultaneously by a pair of observers, representing a spectrum of health care professionals, who scored the children using both the IFS and GCS. Interrater reliability for each pair of observers for each scale was assessed using the kappa statistic. A second series of 10 infants in the intensive care unit with specific diagnoses of acute traumatic or hypoxic/ischemic brain injury were similarly assessed. In the 75 hospitalized infants with a variety of diagnoses, interrater reliability for the GCS was in the "almost perfect," "slight," and "fair" range for the eye-opening, motor, and verbal subtests, respectively. In contrast, the IFS showed interrater reliability in the "almost perfect," "substantial," and "almost perfect" ranges for the three subtests. When applied to infants in an intensive care unit with acute traumatic brain injury or hypoxia/ischemia, the GCS interrater reliability scores were in the "fair" range, while the IFS scores were in the "almost perfect" range. The IFS demonstrates improved interrater reliability in direct comparison to the GCS, particularly in the "verbal/face" component where most pediatric coma scales are deficient. The IFS may prove to be a simple and practical bedside index of brain injury severity in children less than two years of age. PMID- 11011814 TI - Inhibition of neocortical plasticity during development by a moderate concussive brain injury. AB - To determine if a moderate traumatic brain injury (TBI) sustained early in life alters the capacity for developmental plasticity, 17-20-day-old rat pups received a lateral fluid percussion and then reared in an enriched environment for 17 days. Compared to sham-injured controls, this moderate TBI prevented the increase in cortical thickness (1.48 vs. 1.68 mm, p < 0.01) as well as the corresponding enhancement in cognitive performance in the Morris Water Maze (39 vs. 25 trials to criterion, p < 0.05). These injured animals exhibited no significant neuronal degeneration and no evidence of neurologic or motor deficits. These findings strongly support the conclusion that a diffuse brain injury is capable of inhibiting both anatomical and cognitive manifestations of experience-dependent developmental plasticity. PMID- 11011815 TI - Role of nociceptin/orphanin FQ in age-dependent cerebral hemodynamic effects of brain injury. AB - This study was designed to compare the role of the newly described endogenous opioid nociceptin/orphanin FQ (NOC/oFQ) in the reductions of cerebral blood flow (CBF) and pial artery diameter observed following fluid percussion brain injury (FPI) in chloralose anesthetized newborn and juvenile pigs as a function of time postinsult. FPI elevated CSF NOC/oFQ concentration from 70 +/- 3 to 444 +/- 51 within 1 h and to 1,931 +/- 112 pg/mL (n = 7) within 8 h, whereas concentrations returned to control value within 168 h in the newborn. In contrast, FPI elevated CSF NOC/oFQ from 77 +/- 4 to 202 +/- 16 pg/mL (n = 7) within 1 h, while values returned to control value within 8 h in the juvenile. Topical NOC/oFQ (10(-8), 10(-6) M) induced vasodilation was reversed to vasoconstriction by FPI in the newborn while such responses were only attenuated in the juvenile at 1 h post insult (control, 9 +/- 1 and 16 +/- 1%; FPI newborn, -8 +/- 1 and -14 +/- 1%; FPI juvenile, 2 +/- 1 and 5 +/- 1%, n = 7). Such altered dilation returned to control value within 168 h in newborns and 8 h in juveniles. Blood flow in the cerebrum was reduced from 57 +/- 4 to 23 +/- 3 mL x min(-1) x 100 g(-1) (n = 7) within 1 h and returned to control value with 168 h post FPI in newborns. In animals pretreated with [F/G] NOC/oFQ (1-13) NH2 (1 mg/kg, i.v.), a NOC/oFQ antagonist, however, CBF only fell to 39 +/- 4 mL x min(-1) x 100 g(-1) (n = 7) at 1 h post insult in newborns. In contrast, CBF was only reduced from 57 +/- 6 to 32 +/- 2 in untreated and to 39 +/- 3 mL/min(-1) x 100 g(-1) (n = 7) in treated juveniles within 1 h post FPI. Similar observations for reductions in pial artery diameter were made in untreated and treated newborns and juveniles. These data suggest that an elevated CSF NOC/oFQ concentration and altered vascular responsiveness to this opioid contribute to reductions in CBF and pial artery diameter observed following FPI. Because such NOC/oFQ changes were greater in newborns versus juveniles, these data further suggest that NOC/oFQ contributes to age-related cerebral hemodynamic differences in the effects of FPI. PMID- 11011816 TI - Spinal cord injury "outliers": an analysis of etiology, outcomes, and length of stay. AB - Spinal cord injury (SCI) requires significant and expensive medical intervention, including prolonged hospitalization and intense in-patient treatment and rehabilitation. Development of predictive models for lengths of stay in spinal cord injury patients provides a method for early prediction of patients that will require greater care, incur greater costs, and need more intensive medical and rehabilitative services. Early identification of SCI patients at high risk of extended lengths of stay will also allow physicians to treat those patients more aggressively, and permit families, as well as sponsors, to estimate the costs of long-term care. Results of a forward-conditional stepwise multiple logistic regression indicate that the model including age at injury, number of days to rehabilitation admission, number of pressure ulcers, number of medical complications, level of injury, and sponsor of initial hospitalization significantly (chi2 = 220.063, p < 0.001) predicts outliers in terms of rehabilitation length of stay. Overall, the percentage of persons who were correctly classified by the multivariate model was 96.92% (chi2 = 66.85, p < 0.001). The correct prediction rate for outliers was 46% and nonoutliers 97%. This model provides a tool that can be used by health providers, sponsors and patients to aid in the identification of individuals with SCI that may require extended lengths of stay. Today, unfortunately, our health care system is burdened by the heavy influence of economics rather than functional outcome. We maintain that implementation of this model will help to improve care of the SCI patient by early identification of those in need of more extensive resources and improve the economic efficiency needed to provide maximal functional outcome. PMID- 11011817 TI - Riluzole and methylprednisolone combined treatment improves functional recovery in traumatic spinal cord injury. AB - The potential use of riluzole (a glutamate release inhibitor) alone or in combination with methyl-prednisolone (MP) in treating acute spinal cored injury (SCI) was examined. Rats received a contusion injury to the spinal cord using the NYU impactor and were treated with vehicle, riluzole (8 mg/kg), MP(30 mg/kg), or riluzole + MP at 2 and 4 h following injury. Animals continued to receive riluzole treatment (8 mg/kg) for a period of 1 week. The animals were then tested weekly for functional recovery using the BBB open field locomotor score. At the end of testing (6 weeks after injury), each spinal cord was examined for the amount of remaining tissue at the injury site and a myelination index was used to quantify remaining axons in the ventromedial white matter. In this study, only the combination treatment was found to significantly improve behavioral recovery as assessed using the BBB open field locomotor scale. In addition, the combination treatment promoted tissue sparing at the lesion epicenter, but had no clear effect on the index of myelination. The results of this study clearly demonstrate the potential beneficial effects of a combination approach in the treatment of traumatic SCI. PMID- 11011818 TI - Schwannosis: role of gliosis and proteoglycan in human spinal cord injury. AB - Schwannosis (aberrant proliferation of Schwann cells and nerve fibers) has been reported following spinal cord injury (SCI). In this study, we examined the incidence of schwannosis following human SCI, and investigated its relationship to gliosis. We found evidence of schwannosis in 32 out of 65 cases (48%) of human SCI that survived 24 h to 24 years after injury; this incidence rose to 82% in those patients who survived for more than 4 months. Schwannosis was not observed in cases that survived less than 4 months after injury. In affected cases, it was generally noted in areas that had low immunoreactivity for glial fibrillary acidic protein (GFAP), suggesting that reduced gliosis might have contributed to the aberrant proliferation of Schwann cells following SCI. Since chondroitin sulfate proteoglycan (CSPG) has been proposed to play a role in Schwann cell/glial interaction, we performed immunohistochemical staining for CSPG to investigate its potential relationship with schwannosis. CSPG in the injured cord was generally associated with the blood vessel walls, but was also sometimes noted in reactive astrocytes. In SCI with schwannosis, CSPG staining was more prominent and confined largely to the extracellular matrix and basal lamina of proliferating Schwann cells. Our study suggests that Schwann cells, which may have been displaced from spinal roots and introduced into the injured cord through a break in the pial surface, are capable of proliferating and producing CSPG, particularly in the setting of reduced gliosis. Since CSPG has been associated with inhibition of neurite outgrowth, its increased production by aberrant Schwann cells may impair spinal cord regeneration after injury. PMID- 11011819 TI - Effect of liposome-mediated macrophage depletion on Schwann cell proliferation during Wallerian degeneration. AB - The axolemma is considered a well-established mitogen, responsible for Schwann cell proliferation during Wallerian degeneration in the peripheral nerve. However, very little is known about the role of macrophages in Schwann cell proliferation. To test the possible influence of macrophages on Schwann cell proliferation during Wallerian degeneration, macrophages were depleted by dichloromethylene diphosphonate (CI2MDP)-containing liposomes in two-month old C57BL/6J mice. CI2MDP-containing liposomes were injected into the mice intravenously prior to inducing Wallerian degeneration. The injection was repeated every other day to maintain macrophage depletion. Physiologic saline was injected into the control mice. To assess macrophage depletion in vitro, cells were isolated from sciatic nerves at 1, 2, 3, 5, and 7 days post-transection (DPT) and Mac-1 positive cells attached to coverslips were counted. In an in vivo study, Mac-1 positive cells were counted on sciatic nerve sections at the same time points. Throughout the course, the number of Mac-1 positive cells in macrophage-depleted mice was less than that in the control mice both in vivo and in vitro. Schwann cell proliferation was assessed by an in vitro system that reflects in vivo status at the time of cell isolation. Schwann cells were isolated from sciatic nerves at the same time points and proliferation rate was measured by thymidine autoradiography. The proliferation rate was mildly suppressed in macrophage-depleted mice, especially for the initial 3 DPT; however, the pattern of proliferation was not significantly different from controls. These results suggest that macrophages contribute to Schwann cell proliferation during Wallerian degeneration however, their contribution may be relatively limited. PMID- 11011820 TI - The clinical features of severe community-acquired pneumonia presenting as septic shock. Norasept II Study Investigators. AB - PURPOSE: The purpose of this article was to determine the outcome, clinical and prognostic features, and microbiology of a large group of patients with community acquired pneumonia (CAP) presenting in septic shock. MATERIALS AND METHODS: The placebo limb of the Norasept II database was examined. Data were collected on patients in septic shock with a diagnosis of CAP who presented to a participating site from home. RESULTS: One hundred and forty-eight patients met the study criteria. The 28-day survival was 53%. One hundred and four pathogens were isolated from 77 (52%) patients with 24 (16%) patients having polymicrobial infections. The most common pathogen was Streptococcus pneumoniae (19%), followed by Staphylococcus aureus (18%), Haemophilus influenzae (14%), Klebsiella pneumoniae (11%), and Pseudomonas aeruginosa (7%). Infection with P aeruginosa or Acinetobacter species carried a very high mortality (82%). The only clinical variables recorded in the database that could identify patients with pseudomonas or acinetobacter infection was a history of alcohol abuse. Comorbidities were present in 74% of patients, involving predominantly the cardiorespiratory system. Logistic regression analysis demonstrated APACHE II score and serum interleukin 6 (IL-6) concentration to be significant independent predictors of mortality. Patients with pseudomonas or acinetobacter infection had significantly higher IL 6 levels and significantly lower tumor necrosis factor alpha levels when compared with the rest of the cohort of patients. CONCLUSION: A diverse spectrum of both gram-positive and gram-negative pathogens were implicated in patients with CAP presenting in septic shock, necessitating broad spectrum empiric antimicrobial coverage. This coverage should include antipseudomonal activity, particularly in alcoholic patients. Severity of illness (APACHE II score) and IL-6 levels were important prognostic factors. Infection with P aeruginosa and Acinetobacter species carried a very high mortality. PMID- 11011821 TI - Acute respiratory distress syndrome: resource use and outcomes in 1985 and 1995, trends in mortality and comorbidities. AB - PURPOSE: The purpose of this study was to compare resource consumption and mortality between (ARDS) patients with adult respiratory distress syndrome treated at our center in 1985 (45 patients) and those treated in 1995. MATERIALS AND METHODS: This was a retrospective observational study, considering trauma and nontrauma ARDS separately. We recorded severity index scores (APACHE III), infectious complications and multiorgan failure, intensive care unit (ICU) resource consumption (TISS 28), length of stay, time on mechanical ventilation, and ICU mortality. RESULTS: We found no variation in overall ARDS mortality and no reduction in mortality in the ARDS trauma group (43.5% in 1985 vs. 38.5% in 1995, not significant) but a significant increase in mortality among nontrauma septic ARDS patients (68.2% vs. 82.9%, P < .001), largely attributable to the new comorbidities of human immunodeficiency virus (HIV) infection and hematologic malignancy. TISS-28 showed an overall reduction over this time period (49.7 +/- 6.6 vs. 38.3 +/- 9.7, P < .001), due to fewer monitoring measures, particularly a lower use of pulmonary artery catheter. There were no overall changes in length of stay or days on mechanical ventilation between 1985 and 1995, but these variables did increase among the trauma subgroup. CONCLUSION: In our setting, mortality remained constant from 1985 to 1995 among ARDS trauma patients but not among nontrauma ARDS patients because of the new case-mix of the latter population, which now includes HIV and other immunodepressed patients. PMID- 11011822 TI - Influence of amrinone on intestinal villus blood flow during endotoxemia. AB - PURPOSE: The objective of this study was to determine the effects of a continuous infusion of the phosphodiesterase (PDE) inhibitor amrinone on mucosal villus blood flow in a normotensive model of endotoxemia. MATERIALS AND METHODS: Twenty four anesthetized and ventilated rats underwent laparotomy, and an ileal portion was exteriorized and opened by an antimesenteric incision. The ileal segment was fixed on a plexiglass stage with the mucosal surface upward. Microcirculatory parameters were assessed by intravital videomicroscopy. The animals were randomly assigned to receive one of three treatments: infusion of Escherichia coli lipopolysaccharides (LPS, 2 mg/kg/h) without phosphodiesterase inhibitor pretreatment (LPS group); or infusion of LPS with amrinone pretreatment (40 microg/kg/min, start 30 minutes before LPS infusion) (amrinone group), or infusion of equivalent volumes of NaCl 0.9% (control group). Macrohemodynamic parameters (MAP, HR) and microhemodynamic parameters of ileal mucosa (mean diameter of central arterioles = D(A) and mean erythrocyte velocity within the arterioles = VE) were measured 30 minutes before and at 0, 60, and 120 minutes after induction of endotoxemia. Mucosal villus blood flow was calculated from D(A) and VE. RESULTS: In this normotensive endotoxemia model, MAP remained stable in the control and the LPS group but significantly decreased in the amrinone group.The endotoxin-induced decrease of V(E) and D(A) of central arterioles of mucosal villi could be attenuated and prevented, respectively. Thus, the endotoxin-induced decrease of mucosal villus blood flow was diminished but not fully restored by amrinone infusion. CONCLUSION: Our results indicate that amrinone during an early stage of sepsis is of limited value. It attenuates mucosal hypoperfusion but contributes to systemic hypotension. PMID- 11011823 TI - Partial liquid ventilation shows dose-dependent increase in oxygenation with PEEP and decreases lung injury associated with mechanical ventilation. AB - PURPOSE: The purpose of this article is to evaluate the effect of positive end expiratory pressure (PEEP) during partial liquid ventilation (PLV) and to investigate if lung damage associated with mechanical ventilation can be reduced by PLV. MATERIALS AND METHODS: Twenty-two New-Zealand white rabbits were ventilated in pressure-controlled mode maintaining constant tidal volume (10 mL/kg). Lung injury was induced by repeated saline lavage (PaO2 < 100 mm Hg). Two incremental PEEP steps maneuvers (IPSMs) from 2 to 10 cm H2O in 2 cm H2O steps were performed sequentially. The control group received the first IPSM in the supine position and were turned prone for the second IPSM. In the PLV group (n = 7), 12 mL/kg of perfluorodecalin was instilled after lung injury before the two IPSMs. The early prone group (n = 7) received both IPSMs in the prone position. Parameters of gas exchange, lung mechanics, and hemodynamics as well as pathology were examined. RESULTS: During the first IPSM, the PLV group showed a significant increase in PaO2 after instillation of perfluorodecalin (P < .05) and then showed a dose-dependent increase in PaO2 with PEER. The control and EP groups showed improvement in PaO2 only at higher PEEP, eventually showing no intergroup differences at PEEP of 10 cm H2O. During the second IPSM only the PLV group retained its ability to increase PaO2 to the level obtained during the first IPSM (P < .05 compared with control and EP groups). During the first IPSM all three groups showed increasing trend in static compliance (Cst) with PEEP peaking at PEEP of 8 cm H2O. During the second IPSM, only the PLV group showed increase in static compliance with PEEP (P < .05 compared with other groups). Lung histology revealed significantly less hyaline membrane formation in the PLV group (P < .05). CONCLUSION: PLV shows dose-dependent increase in oxygenation with PEEP and may reduce lung damage associated with mechanical ventilation. PMID- 11011824 TI - Effects of milrinone on platelet aggregation in swine with pulmonary hypertension. AB - PURPOSE: The purpose of this study was to investigate whether the effect of milrinone on platelet aggregation was related to the selectivity of vasodilation vasculature in a swine model with PH. MATERIALS AND METHODS: To induce pulmonary hypertension, we injected two sets of acid-washed glass beads in 15 swine, which were divided into two groups (those receiving milrinone or not) and compared with each other. RESULTS: The induction of pulmonary hypertension decreased the platelet count and increased the plasma levels of thromboxane B2 and 6-keto prostaglandin F1alpha. CONCLUSION: A locally high concentration of prostaglandin I2, at least in part, may produce selectivity of vasodilation in the pulmonary vasculature. PMID- 11011825 TI - Measurement of soluble Fas antigen and ligand in circulating serum and intra abdominal or cerebrospinal fluid during gastrointestinal or cerebrovascular surgery. AB - PURPOSE: Soluble Fas antigen (sFas) and ligand (sFasL), which are associated with apoptosis, have not been evaluated in gastrointestinal or cerebrovascular surgery. The aim of this study was to measure these substances in serum, intra abdominal fluid, or cerebrospinal fluid, and to speculate a pathophysiologic role for Fas-FasL apoptosis in surgery. MATERIALS AND METHODS: Arterial blood and intraabdominal or cerebrospinal fluid were collected at intervals from 27 gastrointestinal surgical patients, 10 cerebrovascular surgical patients, and 10 spinal anesthesia patients. RESULTS: Serum sFas levels did not change during and after surgery. Intra-abdominal and cerebrospinal sFas levels were identical to and lower than those in serum. Serum sFasL levels did not change during surgery, but decreased after surgery. Intra-abdominal and cerebrospinal sFasL levels were higher than and identical to those in serum. In spinal anesthesia, cerebrospinal sFas and sFasL levels were lower than those in serum. CONCLUSIONS: Serum sFasL decreases after surgery, whereas intra-abdominal or cerebrospinal sFasL increases, although sFas in each site does not change, suggesting redistribution of activated lymphocytes into local surgical wounds and induction of apoptosis in this site. PMID- 11011826 TI - Personality disorders, history of trauma, and posttraumatic stress disorder in subjects with anxiety disorders. AB - The purpose of the study was to examine the association of personality disorders, history of trauma, and posttraumatic stress disorder (PTSD) in a large sample of subjects with anxiety disorders. Categorical and continuous indices of personality disorders were compared in three groups from the Harvard/Brown Anxiety Disorders Research Project (HARP): subjects with no history of trauma (n = 403), subjects with a history of trauma but no history of PTSD (n = 151), and subjects with a current or past diagnosis of PTSD (n = 68). Subjects with PTSD were more likely to meet criteria for borderline or self-defeating personality disorder than subjects in the other two groups. PTSD subjects also had higher scores on the continuous measures (total number of criteria met) for borderline and self-defeating personality disorder than the other two groups. The findings suggest that a diagnosis of PTSD rather than a history of trauma is associated with borderline and self-defeating personality disorder features. Alternative conceptualizations of axis II features in individuals with PTSD are discussed. PMID- 11011827 TI - Computerized measurement of cognitive impairment and associated neuropsychiatric dimensions. AB - This study aimed to cross-validate the capacity of a computer software program to detect and measure, using a measurement method applied to the content and form analysis of 5-minute speech samples, cognitive impairment and associated comorbid neuropsychiatric psychobiological dimensions in drug-abusing patients. At the University of California-Irvine (UCI) Neuropsychiatric Center, 28 drug-abusing inpatients using illegal drugs were clinically evaluated. Their scores for cognitive impairment derived by the computerized content analysis method were compared with scores derived from selected tests from the Halstead-Reitan Neuropsychological Test Battery, the computerized Automated Neuropsychological Assessment Metric Battery (ANAM), the Alzheimer's Disease Assessment Scale Cognitive Portion, the Stroop Color and Word Test, the Symbol Digit Modalities Test, and the Controlled Oral Word Association Test. The statistical significance (P value) of the correlations of scores from these different measures with scores obtained from the computerized content analysis measures was less than .05 to .001. The comparative "hit rate," detecting cognitive impairment above the norms for each measure administered to these drug-abusing patients, for the computerized content analysis measures and some of the ANAM neuropsychological measures was 75% to 89%, and for the other neuropsychological measures, 25% to 64%. In conclusion, the computerized content analysis methodology applied to 5 minute verbal samples is a valid, rapid, easily administered measurement instrument for assessing the magnitude of cognitive impairment and comorbid neuropsychiatric dimensions. PMID- 11011828 TI - Comorbid psychiatric diagnosis and long-term drinking outcome. AB - This longitudinal study of alcoholics investigated which psychiatric comorbidities among alcoholics would predict very long-term drinking outcome. Previous research has yielded inconsistent findings. We hypothesized that antisocial personality characteristics alone among psychiatric comorbidities would show an association with poorer drinking outcome. The use of multiple measures of psychopathology, a relatively large sample size, and an absence of systematic treatment matching to particular patient groups were all aspects of the current study which allowed for a comprehensive examination of this issue. The study used single and multivariate correlational analyses. The setting was an inpatient Veterans Administration alcohol dependence treatment unit and follow-up clinic. Participants were 255 adult male veterans diagnosed with alcohol dependence. The predictors were the Symptom Checklist 90 (SCL), Minnesota Multiphasic Personality Inventory (MMPI), and Psychiatric Diagnostic Interview (PDI). The outcome measure was the Clinician Rating of Drinking Scale (CRDS). The study showed that antisocial personality characteristics alone were consistently associated with a worse long-term drinking outcome. However, despite the consistent presence of a statistical association between antisocial personality characteristics and a poorer long-term drinking outcome, the small size of the relationship is a very important issue which is discussed in detail. PMID- 11011829 TI - Bipolar II disorder and comorbidity. AB - The validity and reliability of the diagnosis of bipolar II disorder has been questioned by means of comorbidity with nonaffective disorders, including substance abuse, personality disorders, and anxiety disorders. This study examined the comorbid diagnosis of a sample of bipolar II patients, comparing patients with comorbidity and those with "pure" bipolar II disorder. Forty Research Diagnostic Criteria (RDC) bipolar II patients were assessed by means of the Schedule for Affective Disorders and Schizophrenia, Lifetime Version (SADS-L) and Structured Clinical Interview for DSM-III-R axis I (SCID-II) for personality disorders. Patients fulfilling RDC criteria for any psychiatric disorder (except personality disorders) or DSM-IV criteria for any personality disorder were compared with patients without comorbidity. For practical reasons, cyclothymia was not considered as a comorbid diagnosis. Half of the sample had lifetime comorbidity with other psychiatric disorders, mainly personality disorders (33%), substance abuse or dependence (21%), and anxiety disorders (8%). However, only the rates of suicidal ideation (74% v 24%, chi square [chi2] = 9.03, P = .003) and suicide attempts (45% v 5%, chi2 = 8.53, P = .003) were significantly different between patients with and without comorbidity. In summary, although the rates of comorbidity are relatively high in bipolar II disorder, most clinical and course variables are strikingly similar in patients with and without comorbidity except for suicidal behavior, suggesting that comorbidity does not reduce the validity of the diagnosis of bipolar II disorder. PMID- 11011830 TI - Psychosocial issues for women with trichotillomania. AB - Trichotillomania, the compulsive urge to pull out one's hair, is a poorly studied chronic condition affecting 2 to 8 million people, 90% of whom are women. Significant gains have been made about the nature and clinical presentation of this disorder, and yet conceptualizations of trichotillomania remain inconsistent and controversial. Consequently, treatments for trichotillomania have proven to be largely ineffective. Anecdotal case studies have provided us with preliminary data about the impact of this condition on emotional, psychological, and social well-being. To date, there are no empirical studies that focus on patients' subjective experiences with their disorder. The purpose of this report is to identify and discuss the concerns of women with trichotillomania. We have also provided clinical examples to highlight how these concerns are evident in the lives of these women. A total of seven women participated in focused interviews which asked them to reflect on their experiences with compulsive hair pulling. The interview transcripts were analyzed for themes using techniques from the constant comparative method. Additionally, women were asked to complete a demographic self-report. Our analysis identified themes pertinent to negative affects, control, and triggering precipitants. We argue that identifying and integrating these themes in current treatment protocols is the first step to improving the efficacy of treatment for trichotillomania. PMID- 11011831 TI - Alexithymia, gender, and hemispheric functioning. AB - It has been hypothesized that alexithymia is related to an impairment of the right hemisphere or a deficiency in interhemispheric transfer. We used the Toronto Alexithymia Scale-20 (TAS-20) and the tactile finger localization task of Zeitlin et al. to test these relationships on nonclinical samples of college men and women, and also considered the role of short-term memory. Among 47 men, the TAS-20 facets of difficulty identifying feelings or difficulty describing feelings were correlated with poorer performance by the right compared with the left hemisphere in uncrossed trials and poorer interhemispheric transfer of information on crossed trials; short-term memory was not related. Thus, both hemispheric hypotheses were supported for men. However, among 58 women, alexithymia was completely unrelated to either index of hemispheric functioning; instead, poorer short-term memory (specifically digits backwards) strongly predicted poorer interhemispheric transfer. We conclude that deficiencies in right hemisphere function and interhemispheric transfer may contribute to alexithymia in men, but not in women. PMID- 11011832 TI - The impact of parental death versus separation from parents on the mental health of Israeli adolescents. AB - Early research suggested that the loss of a parent during childhood can lead to depression and other psychopathology in children and adults. More recent research has differentiated between loss due to death and separation from parents and has questioned the link between early parental death and psychopathology. We examined the hypothesis that separation from parents has a more detrimental effect than the death of a parent on the mental health of adolescents in the community. Israeli adolescents (N = 844) with a mean age of 16.7 +/- 1.0 years (mean +/- SD) participated in this study. Seventy reported that a parent did not live at home during one of three 5-year periods until they reached the age of 15; 37 reported the death of a parent and 777 were from intact families. All participants completed the Brief Symptom Inventory (BSI), General Well Being Schedule (GWB), Parental Bonding Instrument (PBI), and Perceived Social Support Family/ Friends Scales (PSS-Fam and PSS-Fr). Adolescents who had experienced separation from parents had more psychiatric symptoms (BSI), expressed a lower sense of well being (GWB), experienced less support from their family (PSS-Fam), and felt less cared for and more controlled by their parents (PBI) compared with those belonging to intact families. The death of a parent was not associated with significantly different scores on these variables. When we controlled for parental bonding (PBI) and perceived social support from family (PSS-Fam), the differences between the separation group and the intact family group were no longer significant. The psychological impact of separation from parents involves a greater risk for psychopathology than the death of a parent. The quality of the relationship with the parents moderates the negative impact of separation from them. PMID- 11011833 TI - Parasuicide, anhedonia, and depression. AB - Previous research has shown that anhedonia characterizes suicide attempters. The present study aimed to replicate this finding using the level of depression as a control. Seventy-three depressed parasuicides, 30 nondepressed parasuicides, and 104 matched controls were assessed on the Revised Physical Anhedonia Scale (PAS). Consistent with previous studies, depressed parasuicides were significantly more anhedonic than controls, but nondepressed parasuicides were not significantly more anhedonic than controls. Moreover, multiple linear regression analysis showed that the PAS score explained a negligible part of the variance in the distinction between parasuicides and controls. Anhedonia in parasuicides constitutes a depressive feature and not a temperamental trait. PMID- 11011834 TI - Phenotypic differences in early- and late-onset obsessive-compulsive disorder. AB - Early-onset forms of many medical diseases have been associated with specific genetic anomalies. To assess the potential marker value of onset age in obsessive compulsive disorder (OCD), we examined and compared the phenotypic characteristics of patients with early and later onset. The study sample included 38 children with DSM-IV OCD and 129 adults 19 years of age or older, 77 of whom reported OCD onset prior to age 18 and 52 of whom reported OCD onset at 18 years of age or older. DSM-IV diagnoses were ascertained for all subjects using an amended version of the Diagnostic Interview for Genetic Studies (DIGS). An initial comparison of children and adults with childhood onset revealed several differences, including an earlier onset of clinically significant symptoms without impairment and earlier onset of DSM-IV OCD, a higher frequency of learning disabilities, and fewer obsessions and compulsions among our child patients. For this reason, subsequent analyses included only adult patients with early and later OCD onset. Nonimpairing symptom onset prior to puberty, a relatively aggressive course, and a greater number of obsessions and compulsions unrelated to the amount of time in illness characterized early-onset OCD. Later onset OCD was characterized by nonimpairing symptom onset during puberty, a static course, and relatively few obsessions and compulsions that were variably related to the amount of time in illness. We conclude that children with OCD and adults with childhood onset differ in their report of clinical characteristics and should be analyzed separately in studies concerning the phenotypic characteristics of OCD. Early- and late-onset forms of OCD appear to be characterized by phenotypic features that have important neurobiologic and perhaps genetic implications. PMID- 11011835 TI - Association between cerebral structural abnormalities and dermatoglyphic ridge counts in schizophrenia. AB - Dermatoglyphic ridge counts (1) reflect ontogenic processes during the second trimester of pregnancy and (2) can be influenced by some of the factors that also affect cerebral development. Therefore, the demonstration of an association between dermatoglyphic and cerebral structural measures in patients with schizophrenia would give credence to the view that the structural brain abnormalities associated with this disorder have their origin early in development. Twenty-eight male subjects with schizophrenia and 19 male controls underwent magnetic resonance imaging (MRI) and dermatoglyphic analysis. The pattern of association between the ab-ridge count and nine MRI features was dissimilar in cases and controls for two measures. Associations between dermatoglyphic features, on the one hand, and the frontal CSF (r = .54, P = .004) and fourth ventricular volume (r = .38, P = .05), on the other, were larger in the cases versus the controls (test for interaction, P = .08 and P = .06, respectively). These findings, while in need of replication, support the view that the cerebral structural abnormalities found in patients with schizophrenia are the result of an early pathologic process affecting the development of fetal ectodermal structures. PMID- 11011836 TI - Impaired Wisconsin card sorting test performance in first-episode schizophrenia: resource or motivation deficit? AB - Patients with first-episode schizophrenia (n = 27) and age- and education-matched healthy controls (n = 27) were administered the standard version of the Wisconsin Card Sorting Test (WCST), the Wechsler Adult Intelligence Scale-Revised (WAIS-R), and the Rorschach according to the Comprehensive System (CS). Schizophrenic patients achieved a significantly lower full-scale IQ and made more perseverative responses and achieved fewer categories on the WCST than the healthy control group. No significant associations were observed between effort or motivation and WCST performance. Schizophrenic patients who made more perseverative responses tended to be impoverished in terms of available resources, and functioned in a simplistic way when attending to details of the stimulus field. First-episode schizophrenics are able to generate motives and initiate goal-directed activity, but some of them fail to achieve their goals because the cognitive abilities and available resources required for effective planning, purposeful action, or effective performance are impaired. PMID- 11011837 TI - Expressed emotion and family distress in relatives of patients with schizophrenia in Japan. AB - We evaluated the association between distress and expressed emotion (EE) in family members of patients with schizophrenia by the General Health Questionnaire (GHQ), the Camberwell Family Interview (CFI), and the Five-Minute Speech Sample (FMSS). The GHQ score was higher in the high-EE group determined by both the CFI and FMSS. The difference in the GHQ score between high-EE and low-EE groups was more marked for the CFI. Family distress is closely associated with the EE classification, but the EE classification by the CFI more markedly reflected family distress versus the FMSS. Even in relatives with low EE, distress was marked, and therefore, coping with mental health in family members is important. PMID- 11011838 TI - Depressive disorders in teenage-onset anorexia nervosa: a controlled longitudinal, partly community-based study. AB - The study objective was to examine the prevalence and course of depressive disorders (DDs) in teenage-onset anorexia nervosa (AN) over a period of 10 years. Fifty-one adolescents with AN and a sex- and age-matched control group (n = 51) were assessed at ages 16, 21, and 24 years. Probands and controls were examined in depth using semistructured and structured interviews. Their parents were interviewed on the occasion of the first examination. DDs were assessed using DSM III-R criteria. Subjects with AN had a greatly increased rate of DDs (85%) of all kinds and at all ages as compared with control subjects. The risk of DD during the follow-up period from 21 up to and including 24 years could be predicted by diagnostic group status and the presence of DD during the period from 16 to 21 years, while the risk of DD during the follow-up period from 16 up to and including 21 years was solely predicted by the presence of AN at age 16 years. Long-term resolution of the eating disorder (ED) was associated with the absence of mood disorder or vice versa. Bipolar disorder (BP) occurred at roughly the expected rate (11%) among subjects (probands and controls) with major depression (MDD). In conclusion, depression is a very common comorbid problem in AN: more than four of five individuals with teenage-onset AN had at least one episode of DSM-III-R depression (MD or dysthymia [DT]) within 10 years after onset of the ED. AN appears to trigger the first episode of depression, but once it is manifest, depression predicts further depressive episodes. PMID- 11011839 TI - Pathways and regulation of homocysteine metabolism in mammals. AB - Two intersecting pathways, the methionine cycle and the transsulfuration sequence, compose the mechanisms for homocysteine metabolism in mammals. The methionine cycle occurs in all tissues and provides for the remethylation of homocysteine, which conserves methionine. In addition, the cycle is essential for the recycling of methyltetrahydrofolate. The synthesis of cystathionine is the first reaction in the irreversible pathway for the catabolism of homocysteine by means of the sequential conversion to cysteine and sulfate. This pathway has a limited distribution and is found primarily in the liver, kidney, small intestine and pancreas. Regulation of homocysteine metabolism is achieved by changes in the quantity of homocysteine distributed between the two competing pathways. Two mechanisms are basic to the regulatory process. Changes in tissue content of the relevant enzymes are the response to sustained perturbations. The inherent kinetic properties of the enzymes provide an immediate response to alterations in the tissue concentrations of substrates and other metabolic effectors. S adenosylmethionine, S-adenosylhomocysteine, and methyltetrahydrofolate are of particular importance in that context. PMID- 11011840 TI - Consequences of homocysteine export and oxidation in the vascular system. AB - The risk for arteriosclerosis and thrombosis of patients with severe hyperhomocysteinemia is reduced by homocysteine-lowering therapy. Whether this is the case in patients with mild hyperhomocysteinemia remains to be proved. Another challenge for researchers is to establish a satisfying pathological mechanism of the vasotoxicity of a disturbed homocysteine metabolism. Unfortunately, most in vitro studies use physiologically irrelevant concentrations or forms, or both, of homocysteine. The role of the different oxidized and reduced forms of homocysteine in its metabolism has gained little attention. In the cell, homocysteine is mainly present in its reduced form. In this article export of homocysteine out of the cell is reported to be regulated by a "reduced homocysteine carrier." In vitro endothelial cells export homocysteine at a constant rate in a folate dose-dependent matter. Even at high-normal folate levels, endothelial cells export homocysteine. As soon as homocysteine is exported out of the cell, it will be oxidized to a disulfide with any compound containing a thiol function or undergo a disulfide exchange reaction, both resulting in formation of disulfides of homocysteine. Consequently, in plasma, about 99% of homocysteine is bound to disulfides. Before homocysteine can be metabolized, it needs to be taken up by the cell via carriers, channels, or receptors recognizing the different homocysteine disulfides. In the cell, the homocysteine disulfides are reduced, liberating homocysteine in its reduced form. Next, homocysteine can be metabolized after binding to the homocysteine converting enzymes. In particular, the liver and kidney supposedly take up and metabolize significant amounts of homocysteine. PMID- 11011841 TI - Assay methods for the measurement of total homocyst(e)ine in plasma. AB - Analytical methods to measure plasma total homocyst(e)ine (tHcy) concentrations are reviewed. High-pressure liquid chromatography (HPLC) with fluorometric detection is the most widely used method to determine plasma tHcy concentrations. Both monobromobimane and ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate (SBD-F) are popular thiol-specific fluorogenic agents suitable for tHcy analysis. Monobromobimane has the advantage that it is highly reactive towards thiols, but its hydrolysis products are also fluorogenic, thus necessitating complex chromatography to obtain satisfactory separation between the compounds of interest and interferents. SBD-F does not show fluorescence, thus allowing isocratic separation of SBD-F derivatized thiols. SBD-thiol adducts are light sensitive and require protection against light to ensure reliable results. HPLC with electrochemical detection is also often used and has the advantage that no derivatization of thiols is required prior to detection. A recently reported liquid chromatography electrospray tandem mass spectrometric assay has the potential to become the reference method for plasma tHcy analysis. Other methods to measure plasma tHcy concentrations include gas-liquid chromatography, capillary electrophoresis, and immunoassays. Fluorescence polarization immunoassay compares well with gas chromatography-mass spectrometry and may become the method of choice in routine diagnostic clinical chemistry laboratories. Instability of tHcy in whole blood as well as postprandial and orthostatic variation are preanalytical factors that should be accounted for in plasma tHcy analysis. Between-method and between-laboratory variations in serum tHcy analysis are not yet satisfactory; certified reference material and standardization of the plasma tHcy assay will be essential to reduce between laboratory bias. PMID- 11011842 TI - Homocysteine, coagulation, platelet function, and thrombosis. AB - Over the last 30 years, a growing body of evidence has documented the role of hyperhomocysteinemia (HHcy) as an independent vascular risk factor. However, the mechanisms through which elevated circulating levels of homocysteine (Hcy) cause vascular injury and promote thrombosis remain elusive. Most findings have been achieved in in vitro studies employing exceedingly high concentrations of Hcy, whereas only a few studies have been carried out in vivo in humans. In homocystinuric patients, homozygotes for mutations of the gene coding for the cystathionine beta-synthase enzyme, abnormalities of coagulation variables reflecting a hypercoagulable state, have been reported. In vitro studies provide a biochemical background for such a state. In homocystinuric patients, an in vivo platelet activation has also been reported. The latter abnormality is not corrected by the bolus infusion of concentrations of hirudin, which determines a long-lasting impairment of the conversion of fibrinogen to fibrin by thrombin; in contrast, it appears at least in part lowered by the administration of the antioxidant drug probucol. During the autooxidation of Hcy in plasma, reactive oxygen species are generated. The latter initiate lipid peroxidation in cell membranes (potentially responsible for endothelial dysfunction) and in circulating lipoproteins. Oxidized low-density lipoproteins (LDL) may trigger platelet activation as well as some of the hemostatic abnormalities reported in such patients. Thus the oxidative stress induced by Hcy may be a key process in the pathogenesis of thrombosis in HHcy. Accumulation of adenosylhomocysteine in cells (a consequence of high circulating levels of homocysteine) inhibits methyltransferase enzymes, in turn preventing repair of aged or damaged cells. This mechanism has been recently documented in patients with renal failure and HHcy and provides an additional direction to be followed to understand the tendency to thrombosis in moderate HHcy. PMID- 11011843 TI - Genetic modulation of homocysteinemia. AB - With the identification of hyperhomocysteinemia as a risk factor for cardiovascular disease, an understanding of the genetic determinants of plasma homocysteine is important for prevention and treatment. It has been known for some time that homocystinuria, a rare inborn error of metabolism, can be due to genetic mutations that severely disrupt homocysteine metabolism. A more recent development is the finding that milder, but more common, genetic mutations in the same enzymes might also contribute to an elevation in plasma homocysteine. The best example of this concept is a missense mutation (alanine to valine) at base pair (bp) 677 of methylenetetrahydrofolate reductase (MTHFR), the enzyme that provides the folate derivative for conversion of homocysteine to methionine. This mutation results in mild hyperhomocysteinemia, primarily when folate levels are low, providing a rationale (folate supplementation) for overcoming the genetic deficiency. Additional genetic variants in MTHFR and in other enzymes of homocysteine metabolism are being identified as the cDNAs/genes become isolated. These variants include a glutamate to alanine mutation (bp 1298) in MTHFR, an aspartate to glycine mutation (bp 2756) in methionine synthase, and an isoleucine to methionine mutation (bp 66) in methionine synthase reductase. These variants have been identified relatively recently; therefore additional investigations are required to determine their clinical significance with respect to mild hyperhomocysteinemia and vascular disease. PMID- 11011844 TI - Biological and environmental determinants of plasma homocysteine. AB - This article gives an overview over common physiological, lifestyle, and pathological conditions that may modulate the homocysteine status. The interplay of several environmental factors, including age, gender, nutrition, smoking, and coffee consumption and physical activity with commonly used drugs and prevalent diseases are described. In most cases, an abnormal homocysteine status is not caused by a single factor alone but often is the result of combined effects. We address these frequently found "clusters" of homocysteine-modulating factors. Finally, we give an overview of likely causes of hyperhomocysteinemia found in an authentic material. This material is based on 2462 routine measurements of plasma total homocysteine carried out at the Haukeland University Hospital. The data represent the total number of combined homocysteine and methylmalonic acid determinations, requested by general practitioners in Norway during February 1998. PMID- 11011845 TI - Hyperhomocysteinemia, vascular pathology, and endothelial dysfunction. AB - Hyperhomocysteinemia has been associated with premature atherothrombotic vascular disease. It is not known whether hyperhomocysteinemia induces a distinct type of vascular disease. Its interaction, if any, with traditional risk factors also remains unclear. The pathophysiological mechanisms linking hyperhomocysteinemia to vascular disease have been extensively studied in vitro and in animals. From these studies, it has been suggested that homocysteine limits the bioavailability of nitric oxide (NO), increases oxidative stress, stimulates smooth cell proliferation, and alters elastic wall properties. The relevance of these proposed mechanisms in vivo is unclear, because clinical studies have yielded controversial results with regard to the relation between plasma homocysteine levels and indices of endothelial function, such as brachial artery flow-mediated vasodilatation and plasma levels of endothelium-derived marker proteins. Up till now, there have been no controlled data on the effects of homocysteine-lowering treatment on vascular function or clinical end points. The precise mechanisms (if any) by which homocysteine mediates its adverse vascular effects are in fact unknown but may relate to impaired endothelial and smooth muscle cell function. PMID- 11011846 TI - Mild hyperhomocysteinemia is an independent risk factor of arterial vascular disease. AB - Evidence of a positive association between mild hyperhomocysteinemia and arterial vascular disease has been accumulating in the last decade. Mild hyperhomocysteinemia acts as an independent vascular risk factor with equal strength as hypercholesterolemia and smoking. If jointly present with hypertension and smoking, its effect seems synergistic. This could make the outcome of homocysteine-lowering intervention beneficial, particularly in cases with concomitance of conventional vascular risk factors. So far, however, data on the clinical outcome of homocysteine-lowering treatment with a simple, safe, and cheap vitamin regimen are lacking. Trials investigating a beneficial clinical effect of homocysteine-lowering treatment using folic acid in a dose ranging from 0.2 to 5 mg daily, alone or in combination with vitamin B12 with or without vitamin B6 versus placebo, are ongoing. Furthermore, exploration of the unifying mechanism by which increased homocysteine levels may lead to both arterial and venous occlusions is warranted. These lines of investigations have to provide the ultimate proof of causality of hyperhomocysteinemia in vascular disease in the near future. PMID- 11011847 TI - Homocysteine and venous thrombosis: outline of a vitamin intervention trial. AB - In the past years several case-control studies established the association of an elevated plasma homocysteine concentration and the risk of venous thromboembolism. It is still unclear if elevated homocysteine concentrations can cause venous thrombosis. The VITRO (VItamins and ThROmbosis) trial is the first multicenter, randomized, double-blind and placebo-controlled study to evaluate the effect of homocysteine-lowering therapy by means of 5 mg folic acid, 0.4 mg vitamin B12 and 50 mg vitamin B6. The study is a secondary prevention trial in 600 patients who suffered from a first episode of idiopathic deep vein thrombosis (DVT) or pulmonary embolism (PE), or both. There will be 300 hyperhomocysteinemic and 300 normohomocysteinemic patients included, all with an objectivated venous thrombosis. The end point is recurrence of venous thrombosis. PMID- 11011848 TI - Interaction between hyperhomocysteinemia and inherited thrombophilic factors in venous thromboembolism. AB - Hyperhomocysteinemia is an established risk factor for deep vein thrombosis. Factor V Leiden has been reported to potentiate the thrombotic risk related with severe hyperhomocysteinemia, being more represented in thrombotic patients with homocystinuria as compared with patients without a history of thrombosis. The results concerning the interaction between moderate hyperhomocysteinemia and inherited thrombophilic factors such as Factor V Leiden or the prothrombin G20210A mutation are contradictory. The relative risk for venous thrombosis has been reported to be increased 10- to 50-fold in patients carrying both hyperhomocysteinemia and inherited thrombophilia in comparison with normal controls, suggesting a synergistic interaction, yet other studies failed to confirm such conclusion. The heterogeneity of these findings is in part due to the small number of individuals with double defects, leading to statistically unreliable results. Genotyping for mutations that are possible causes of moderate hyperhomocysteinemia, such as the thermolabile variant (C677T) of methylenetetrahydrofolate reductase (MTHFR), does not seem useful to identify individuals at higher risk for venous thromboembolism. In fact, in most of the studies the presence of the C677T MTHFR homozygous genotype does not increase the thrombotic risk associated with Factor V Leiden or the prothrombin mutation. PMID- 11011849 TI - Homocysteine and renal disease. AB - Hyperhomocysteinemia refers to an elevated circulating level of the sulfur containing amino acid homocysteine and has been shown to be a risk factor for vascular disease in the general population. In patients with renal failure, hyperhomocysteinemia is a common feature. The underlying pathophysiological mechanism for this phenomenon is unknown. Proposed mechanisms include reduced renal elimination of homocysteine and impaired nonrenal disposal, possibly because of inhibition of crucial enzymes in the methionine-homocysteine metabolism by the uremic milieu. Absolute or relative deficiencies of folate, vitamin B6, or vitamin B12 may also play a role. Several case-control and prospective studies have now indicated that hyperhomocystenemia is an independent risk factor for atherothrombotic disease in patients with predialysis and end stage renal disease. In renal patients, plasma homocysteine concentration can be reduced by administration of folic acid in doses ranging from 1 to 15 mg per day. In more than 50% of the cases, however, the homocysteine concentration remains above 15 micromol/L. The effects of vitamin B12 or vitamin B6 are unclear. Large intervention trials are now needed to establish whether homocysteine-lowering therapy will reduce atherothrombotic events in patients with renal failure. These studies are now planned or are ongoing. PMID- 11011850 TI - Hyperhomocysteinemia and risk of vascular disease in women. AB - Plasma levels of total homocysteine (tHcy), a possible risk factor for vascular disease, are generally lower in women than in men and lower in premenopausal women than in postmenopausal women. This article reviews studies that have investigated associations of hyperhomocysteinemia with risk of vascular disease among women or that compared risk by stratum of gender or menopausal status. Seven out of 12 epidemiological studies that included both men and women found hyperhomocysteinemia to be a stronger risk factor in women than in men. However, the interaction effect was statistically significant for only 1 study. Three studies observed no risk difference between men and women, and 2 observed a weaker association in women. In addition, 3 studies that consisted (almost) entirely of women observed direct associations of hyperhomocysteinemia with vascular disease risk, comparable to associations observed in male populations. Hyperhomocysteinemia was associated with increased risk in populations of both young and elderly women, but only few studies have compared risks among premenopausal and postmenopausal women. However, the limited data indicate that hyperhomocysteinemia is also associated with elevation of vascular disease risk before the menopause. The stronger association among women in some studies may be explained by aspects of the study design, such as age at inclusion (i.e., women usually suffer from vascular diseases later in life than do men), or aspects of the data analysis, such as use of an overall instead of a gender-specific cutoff point. Of course, one cannot exclude the possibility that women are somehow more susceptible to detrimental effects of tHcy than men are, although there is evidence from other studies that estrogens have a "protective" effect on the vascular wall and a favorable effect on hemostasis. In conclusion, we should consider hyperhomocysteinemia as a potential risk factor for vascular disease in both men and women, before and after the menopause. PMID- 11011851 TI - Vascular complications of severe hyperhomocysteinemia in patients with homocystinuria due to cystathionine beta-synthase deficiency: effects of homocysteine-lowering therapy. AB - Homocystinuria (HCU) due to cystathionine beta-synthase (CBS) deficiency leads to severe hyperhomocysteinemia (HHcy). Vascular events (VE) remain the major cause of morbidity and mortality in the untreated patients with HCU. The study on the natural history of untreated HCU disclosed that, at the time of maximal risk, in other words beyond 10 years old, there was one event per 25 years. Recent studies from Australia (n = 32), The Netherlands (n = 28), and Ireland (n = 24) have documented the effects of long-term treatment on the vascular outcome of a total of 84 patients with 1314 patient-years of treatment for HCU. The mean (range) age was 27.8 (2.5 to 70) years. Five VE were recorded during treatment; one pulmonary embolism, two myocardial infarctions, and two abdominal aneurysms. All five VE occurred in B6-responsive patients at a mean (range) age of 48.8 (30 to 60) years. In 1314 patient-years of treatment, 53 VE would have been expected if they remained untreated; instead only 5 were documented, relative risk = 0.091 (95% confidence interval [CI] 0.043 to 0.190; p < 0.001). Appropriate homocysteine lowering therapy for severe HHcy significantly reduced the vascular risk in patients with HCU. VE were rare with treatment despite the fact that the post treatment homocysteine levels were several times higher than the cutoff point for homocysteine in the normal population. The present findings may have relevance to the current concept of "mild HHcy" as a risk factor for vascular disease, with elevated plasma homocysteine levels considerably lower than that of the post treatment levels in this group of reported patients. PMID- 11011853 TI - Fortifying food with folic acid. AB - Folic acid supplementation may help prevent the following three common and important disorders: neural tube defect pregnancies, ischemic heart disease and strokes, and possibly colon cancer. Several studies have irrefutably established that folic acid supplementation in proper doses reduces neural tube defect pregnancies. Fortified food would be the most reliable way to address this health issue. Several epidemiologic reports have established a close link between hyperhomocysteinemia and ischemic heart disease. This is supported by evidence obtained from genetic defects leading to elevated plasma levels of homocysteine. There is reasonable evidence to conclude that hyperhomocysteinemia is the actual cause of excessive cardiac diseases. Again, folic acid supplementation in proper doses could potentially reduce ischemic heart disease by 40% There is also some evidence to suggest that folic acid treatment reduces the incidence of colon cancer. The reduction in all three health problems is dependent upon the dose of folic acid administered. The levels needed are rarely supplied by the daily food intake, even under ideal conditions, although breakfast cereals address this problem at least in part. It is proposed that flour be fortified because it is a food product heavily used by most people, and that governmental agencies should oversee such programs because they are responsible for setting public health policies. PMID- 11011852 TI - Vitamin supplements and cardiovascular risk: review of the randomized trials of homocysteine-lowering vitamin supplements. AB - Epidemiological studies have shown that higher blood homocysteine levels appear to be associated with higher risks of coronary, cerebral, and peripheral vascular disease and are inversely related to blood levels of folate and of vitamin B12 and vitamin B6. However, observational studies cannot exclude the possibility that elevated homocysteine levels may be associated with some other factor, rather than being causally related to vascular disease. Large-scale clinical trials of sufficient dose and duration of treatment are required to test this hypothesis, but there was substantial uncertainty about the optimal vitamin regimen to test in such trials. A meta-analysis of 12 randomized trials of vitamin supplements to lower homocysteine levels was carried out to determine the optimal dose of folic acid required to lower homocysteine levels and to assess whether vitamin B12 or vitamin B6 had additive effects. This meta-analysis demonstrated that reductions in blood homocysteine levels were greater at higher pretreatment blood homocysteine levels and at lower pretreatment folate concentrations. After standardization for a pretreatment homocysteine concentration of 12 micromol/L and folate concentration of 12 nmol/L (approximate average concentrations for western populations), dietary folic acid reduced homocysteine levels by 25% (95% confidence interval [CI]: 23 to 28%) with similar effects in a daily dosage range of 0.5 to 5 mg. Vitamin B12 (mean 0.5 mg) produced an additional reduction in blood homocysteine of 7%, whereas vitamin B6 (mean 16.5 mg) did not have any significant effect. Hence, in typical populations, daily supplementation with both 0.5 to 5 mg folic acid and about 0.5 mg vitamin B12 would be expected to reduce homocysteine levels by one quarter to one third (from about 12 micromol/L to about 8 to 9 micromol/L). Large-scale randomized trials of such regimens are now required to determine whether lowering homocysteine levels by folic acid and vitamin B12, with or without added vitamin B6, reduces the risk of vascular disease. PMID- 11011854 TI - New international network will study HIV prevention. PMID- 11011855 TI - Dermatofibrosarcoma protuberans: early recognition and treatment. PMID- 11011856 TI - Possible ticlopidine-induced cholestatic jaundice. PMID- 11011857 TI - Vacuum extraction: a necessary skill. PMID- 11011858 TI - Weighing the evidence for vitamin supplementation and CVD prevention. PMID- 11011859 TI - Sickle cell disease in childhood: Part II. Diagnosis and treatment of major complications and recent advances in treatment. AB - Treatment advances over the past 25 years have significantly decreased morbidity and mortality in children with sickle cell disease. Aggressive management of fever, early diagnosis of acute chest syndrome, judicious use of transfusions and proper treatment of pain can improve quality of life and prognosis for these children. Prophylactic hydroxyurea therapy has been shown to reduce the incidence and severity of pain crises in adults with sickle cell disease and has been effective in limited studies conducted in children. Research into stem cell transplantation provides hope that a cure for sickle cell disease may be possible. PMID- 11011860 TI - Assisted vaginal delivery using the vacuum extractor. AB - Vacuum extractors have replaced forceps for many situations in which assistance is required to achieve vaginal delivery. Compared with metal-cup vacuum extractors, soft-cup devices are easier to use and cause fewer neonatal scalp injuries; however, they detach more frequently. Vacuum extractors can cause neonatal injury. These devices should be employed when indicated, usually for a nonreassuring fetal heart tracing or failure to progress in the second stage of labor. Complications may be minimized if the physician recognizes contraindications to the use of vacuum extraction. Complete documentation is essential. PMID- 11011861 TI - Alternative therapies: Part II. Congestive heart failure and hypercholesterolemia. AB - Natural supplements are widely used by the American public but, while claims of their therapeutic effects abound, medical research does not always support their effectiveness. Clinical trials using Q10 for the management of congestive heart failure have had conflicting results; hawthorn is prescribed in Germany for the treatment of this condition, but no trials have been conducted in the United States. Although initial research about the use of garlic in the management of hypercholesterolemia was encouraging, follow-up studies have failed to verify these results. Substituting soy protein for high-fat animal protein diets, however, does have a beneficial effect on serum lipid levels. So far, cholestin (a natural product containing several statins) has proved to be a cost-saving lipid-lowering medication, and fenugreek may offer modest improvement as well. Gugulipid is also promising but requires further research. PMID- 11011862 TI - Proteinuria in adults: a diagnostic approach. AB - Proteinuria is a common finding in adults in primary care practice. An algorithmic approach can be used to differentiate benign causes of proteinuria from rarer, more serious disorders. Benign causes include fever, intense activity or exercise, dehydration, emotional stress and acute illness. More serious causes include glomerulonephritis and multiple myeloma. Alkaline, dilute or concentrated urine; gross hematuria; and the presence of mucus, semen or white blood cells can cause a dipstick urinalysis to be falsely positive for protein. Of the three pathophysiologic mechanisms (glomerular, tubular and overflow) that produce proteinuria, glomerular malfunction is the most common and usually corresponds to a urinary protein excretion of more than 2 g per 24 hours. When a quantitative measurement of urinary protein is needed, most physicians prefer a 24-hour urine specimen. However, the urine protein-to-creatinine ratio performed on a random specimen has many advantages over the 24-hour collection, primarily convenience and possibly accuracy. Most patients evaluated for proteinuria have a benign cause. Patients with proteinuria greater than 2 g per day or in whom the underlying etiology remains unclear after a thorough medical evaluation should be referred to a nephrologist. PMID- 11011863 TI - Management of bipolar disorder. AB - Bipolar disorder most commonly is diagnosed in persons between 18 and 24 years of age. The clinical presentations of this disorder are broad and include mania, hypomania and psychosis. Frequently associated comorbid conditions include substance abuse and anxiety disorders. Patients with acute mania must be evaluated urgently. Effective mood stabilizers include lithium, valproic acid and carbamazepine. A comprehensive management program, including collaboration between the patient's family physician and psychiatrist, should be implemented to optimize medical care. PMID- 11011864 TI - Update on vitamin supplements for the prevention of coronary disease and stroke. AB - Dietary antioxidants and folic acid may play a role in the pathophysiology of coronary disease and stroke. We review patient-oriented evidence on the effectiveness of supplementation with antioxidants and/or folic acid in the prevention of myocardial infarction and stroke. Observational data suggest cardiovascular benefit of vitamin E supplementation, but results of controlled clinical trials are inconsistent regarding the effect on nonfatal myocardial infarction. Moreover, studies have not shown a protective effect of vitamin E against fatal myocardial infarction and have not addressed stroke. For vitamin C and folic acid supplementation, observational data are inconsistent and controlled clinical trials are lacking. Thus, the available evidence is insufficient to recommend the routine use of vitamin E, vitamin C or folate supplements for the prevention of myocardial infarction or stroke. The evidence argues against the use of beta carotene supplements for this purpose. The costs and risks associated with these supplements are low, however, and physicians may choose to recommend vitamin E, folate and/or vitamin C supplementation pending conclusive evidence from clinical trials. PMID- 11011865 TI - ACOG issues recommendations for the management of endometriosis. American College of Obstetricians and Gynecologists. PMID- 11011866 TI - Spatial stimulus-response compatibility and coding of tactile motor events: influence of distance between stimulated and responding body parts, spatial complexity of the task and sex of subject. AB - We studied spatial stimulus response compatibility in the somatosensory modality by instructing 16 men and 16 women to press a key using the left or the right thumb in response to a nonnoxious electric stimulation delivered either to the left or to the right little finger or, in different blocks, to the left or to the right malleolar region. The task was performed in compatible (stimulus and key response on the same side of the corporeal midline) and in incompatible conditions (stimulus and key-response on opposite sides of the corporeal midline). In Exp. 1 subjects were tested while keeping their limbs in anatomic position; in Exp. 2 subjects performed the task while keeping the left upper and lower limbs on the right side and the right limbs on the left side of the bodily midline (crossed position). The compatibility effect was observed in both experiments and was higher for stimuli delivered to the little finger than to the malleolar region. This suggests that the cost of inhibiting compatible responses is maximal when stimulated and responding body parts are contiguous. Moreover, in the spatially most demanding task (Exp. 2) men outperformed women for both speed and accuracy suggesting a sex related specialisation in the spatial processing of somatosensory information. PMID- 11011867 TI - Laser-induced afterimages in humans. AB - Afterimages induced by incoherent light sources have been studied as complex visual perceptions for over 200 years; however, individuals who have viewed a coherent source (a laser) on-axis have reported visual experiences unlike those observed for full-field flashes from incoherent sources. In the present study, 10 volunteers viewed all combinations of blue, green, and red laser light and background colors that matched the wavelengths of the laser sources (a total of 9 conditions). The bright focal (50 microm retinal irradiance diameter) 3-sec. exposures (approximately 9 log trolands-40% of the maximum permissible exposure level) given to the volunteers were administered as they performed a simulator tracking task. A 50/50 mirror (a mirror which reflected 50% of the laser beam into the optical pathway while allowing the 50% of the light from the visual scene to pass through the mirror) permitted simultaneous intrabeam viewing of the laser source and the scene. The volunteers were asked to report what they saw immediately after the laser was turned off and 1-min postexposure. The immediate reports indicated that the image they observed was the same color as the laser source and not the complementary color. Also, the images were of ten surrounded by well-defined borders and, regardless of laser's or background's col or, were generally red. One minute following presentation of the laser light the images seen were predominantly purple, dark or no longer present; however, the edge color when present generally appeared red. The immediate appearance of the images was inferred to be neural in nature and not photochemical. These results suggest the properties of the laser source, i.e., coherency, monochromaticity, and the laser's capacity to place an intense beam of light in a small retinal area, all contributed to the unique appearance of the postflash images. PMID- 11011868 TI - Topology and mental processes. AB - The study reported here considers the effect of rotation on the decision time taken to compare nonrigid objects, presented as like and unlike pairs of knots and unknots. The results for 48 subjects, 21 to 45 years old, support the notion that images which have a characteristic 'foundation part' are more easily stored and accessed in the brain. Also, there is evidence that the comparison of deformable objects is processed by mental strategies other than self-evident mental rotation. PMID- 11011869 TI - Coincidence timing of a soccer pass: effects of stimulus velocity and movement distance. AB - The effect of stimulus velocity and movement extent on coincidence timing and spatial accuracy of a soccer pass was investigated. A Bassin anticipation timer provided light stimulus velocities of 1.79 or 2.68 m/sec. (designated as "Low" and "High", respectively), and subjects were required to kick a stationary soccer ball so that it struck a target in coincidence with the arrival of the light stimulus at the end of the runway. Two kick types were used. The "Short" condition began with the subject 70 cm from the ball and required a single forward step with the nonkicking leg before making the kick. The "Long" condition began 140 cm from the ball and required two steps before the kick. Twenty male subjects were given 16 trials under each of the four combinations of stimulus velocity and kick type. The expectation that the faster stimulus velocity would be associated with lower coincidence timing scores for both absolute error (AE) and variable error (VE) and with late responding for constant error (CEO) was upheld with the exception that for the Long Kick-High Velocity condition, AE was highest. The index of preprogramming (IP) was used to test the hypothesis that a two-stage control process would characterise coincidence anticipation performance involving whole-body movements. Results showed that the preparatory phase of responding produced zero-order IPs signifying reliance on feedback control. Also, while the striking phase produced high IP and suggested reliance on preprogrammed control, the possibility that the High Velocity conditions may have limited the responses was recognised. As a consequence, the role of open-loop processes remained equivocal. The findings are, however, in agreement with the view that the sensorimotor and movement-execution phases of responding require a process that is characterised by adaptability to regulatory features of the environment via closed loop mechanisms involving perception-action coupling. PMID- 11011870 TI - The dominant ear. AB - Presented are some aspects of the dominant ear or earedness which are also important for other lateralities. Earedness refers not only to a sensory but also to a motor phenomenon. PMID- 11011871 TI - Comparison of health promotion and deterrent prompts in increasing use of stairs over escalators. AB - This study compared the effectiveness of two point-of-decision prompts within the same environmental setting. The effects of a health promotion sign were compared with activity change resulting from a deterrent sign. Individuals were observed using the upward stairs or upward escalator at a midwest regional air port during a 5-week period in which intervention signs were compared with no sign conditions on activity choice. During Weeks 1, 3, and 5 behaviors were assessed without any prompts. During Week 2, a health promotion sign was posted at the behavioral choice point which read "Keep your heart healthy, use the stairs." During Week 4, a deterrent sign which read "Please limit escalator use to staff and those unable to use the stairs" was posted at the same behavioral choice point. Younger (<40) women (14.8%) and men (10.8%) used the stairs more frequently, followed by older women (9.3%) and older men (6.9%). A hierarchical log linear analysis showed that stair use increased during both interventions, which was significantly moderated by age. Point-of-decision prompts appear to be effective environmental interventions for promoting increases in physical activity. PMID- 11011872 TI - Satisfaction with life scale: analysis of factorial invariance for adolescents and elderly persons. AB - The purpose of this work is to analyze the factorial invariance of the Satisfaction With Life Scale across samples of adolescents and elderly persons. Data from 266 subjects were analyzed. Half were Spanish junior high-school students (65 girls and 68 boys) and the other half were Spanish elderly people (68 women and 65 men). Single-group analyses showed an acceptable one-factor model for both adolescent and elderly groups. Sequential multigroup analyses to test the equivalence of factor structures for adolescent and elderly groups showed that factor loadings and variances are not invariant. The scale is sensitive to age in these groups. PMID- 11011873 TI - The self-choice elaboration effects on incidental memory of Japanese historical facts. AB - The subjects performed an orienting task involving 3 conditions, followed by unexpected tests which included free recall, name-matching and name-selec tion. Conditions were designed to force self-generated elaboration, self-choice elaboration, and experimenter-provided elaboration. In the self-generated elaboration condition, subjects were presented target sentences, e.g., Nobunaga ODA burned down ENRYAKUJI Temple, and asked to answer an elaborative interrogation, e.g., Why did Nobunaga ODA burn down ENRYAKUJI Temple? about each sentence. In the self-choice elaboration condition, subjects selected one of the alternative answers to an elaborative interrogation about each sentence. In the experimenter provided elaboration condition, subjects were presented an answer which they rated for congruity as the correct answer to the elaborative interrogation. In the free recall test, self generated elaboration led to better performance than the other two conditions for which no difference was observed. However, in the name-matching and name-selection tests, scores were better for self choice elaboration and self-generated elaboration than for experimenter provided elaboration. These results were interpreted as demonstrating that self choice elaboration, in addition to self-generated elaboration, led to effective encoding in memory. PMID- 11011874 TI - Sex differences in sport spectator involvement. AB - Among 107 male and 166 female college students, the present study examined sex differences in involvement with sports as spectators. A 14-item Likert-type scale was used to measure the subjects' emotional, cognitive, and behavioral involvement in sports as spectators. Confirming prior findings, male subjects exhibited higher involvement with sport spectating than did females. The current data also provide support for the unidimensionality, reliability, and validity of the Sport Spectator Involvement Scale. PMID- 11011875 TI - Relationship between preference for red and locus of control. AB - The validity of the assumptions about the color red in the Luscher color theory were examined by correlating preference for red to locus of control and sex. Undergraduates (100 men and 100 women) who scored < or = 8 or > or = 16 on the Rotter Internal-External Locus of Control Scale were administered the short form of the Luscher Color Test. Consistent with the assumptions by Luscher about the color red, the analysis of variance showed that Internal scorers preferred red significantly more than External scorers, but no sex difference was found. PMID- 11011876 TI - Competitive worries, sport confidence, and performance ratings for young swimmers. AB - The purpose of this study was to examine the relationships among competitive worries, sport confidence, and performance of young athletes. Participants were 143 young swimmers aged 11 to 12 years. The athletes completed trait and state questionnaires (competitive worries and sport confidence) in noncompetitive and competitive conditions, respectively. The results indicated: (a) significant relationships among trait and state characteristics and between trait General Self-confidence and performance ratings in both races as well as between performance ratings in Races 1 and 2; (b) significant differences in state variables among athletes with moderate or high and low scores on the trait variables; (c) significant differences in performance among athletes with moderate or high and low scores on the trait General Self-confidence and trait Positive Thinking in Race 1 and on variables trait General Self-confidence, state General Self confidence, and state Positive Thinking Race 2; and (d) trait General Self-confidence and trait Confidence in Unfavorable Situations were the most important predictors of young swimmers' performances. These results may be useful in application to competitive sports for young athletes. They may help in psychodiagnostic procedures and may be used for both the content and the direction of individual programs for psychological preparation of young athletes. PMID- 11011877 TI - Latin square analysis of variance using summary statistics. AB - The method for computing a Latin square analysis of variance using summary statistics is presented. PMID- 11011878 TI - Comparison of water- and land-based exercise in the reduction of state anxiety among older adults. AB - This study examined the effect of an acute exercise program of different exercise conditions on state anxiety among older adults. 73 healthy subjects were recruited and randomly assigned to either water exercise group (n = 36) or land exercise group (n = 37). State anxiety was assessed before exercise and following exercise. Subjects in the water exercise performed a 70-min. exercise program consisting of a 10-min. warm-up, a 20-min. brisk walking, a 20-min. rhythmic dancing, a 10-min. resistance training, and a 10-min. cool down exercise. The land exercise program contained a 10-min. warm up and a 30-min. combined endurance and resistance exercise, followed by a 10-min. cool down exercise. Analysis showed that both exercise groups scored significantly (p<.001) lower on anxiety after exercise, but there was no significant interaction for group by trial for scores on state anxiety. The data indicate that elderly persons who participate in these types of exercise report lower state anxiety. PMID- 11011879 TI - Suicide and occupation in Hong Kong. AB - Suicide rates were highest among Service workers and the lowest in Agriculture workers and fisherfolk in Hong Kong. PMID- 11011880 TI - Effect of exercise-based cardiac rehabilitation on mobility and self-esteem of persons after cardiac surgery. AB - This study adopted an experimental design with using a nonequivalent, posttest only control group to study the rehabilitation outcomes of 152 persons who received cardiac surgery. 37 subjects in a rehabilitation group participated in a 2-mo. exercise-based cardiac rehabilitation programme, and another 115 subjects who did not attend the programme formed the control group. The subjects' self esteem was measured on the Adult Source of Self-esteem Inventory by Elvoson and Fleming, and their mobility skill was measured by a simple mobility test based on New York Heart Association Classification. Analysis of covariance (with covariance analysis of the subjects' age, years of education, occupational skill, and mobility skill) indicated that the experimental group scored higher on positive self esteem and showed significantly better improvement in mobility skill. The exercise-based cardiac rehabilitation programme positively affected physical and psychological outcomes. Also, subjects' self-esteem was significantly correlated with their mobility skills (r=.21, p<.05) among those aged under 60 years (n=96) but not among those aged 60 or above (n=53). PMID- 11011881 TI - Loneliness and adaptation among second-generation Portuguese migrants to France: roles of perceived responsibility and control. AB - This study explores the association of perceived responsibility for loneliness and perceived control over loneliness on social adaptation and psychological adaptation among 109 second-generation Portuguese migrants to France. Analysis showed that rated perceived control was more highly associated with scores on social adaptation and psychological adaptation than perceived responsibility. PMID- 11011882 TI - Incidence of Savant Syndrome in Finland. AB - The general incidence of Savant Syndrome was assessed in Finland. First, a survey was made of all 583 facilities which served people with mental retardation. Second, letters asking for information regarding people with Savant Syndrome were published in two key Finnish journals of the field. We received reports of 45 cases of Savant Syndrome. This makes an incidence rate of 1.4 per 1,000 people with mental retardation. The most common form of exceptional skills was calendar calculation, followed by feats of memory. PMID- 11011883 TI - Stutterers' vocal reaction times to unilaterally presented high and low frequency verbs. AB - The current study investigated vocal reaction times of 11 stuttering participants to verbs with high and low frequencies of occurrence when these verbs were unilaterally visually presented. No significant main effects or interactions between stimuli and visual-half field were found. Reaction times were 16 msec. faster after right visual field presentations and the Pearson correlation coefficient between visual fields .71 (p=.02). These results were interpreted as suggesting that the left hemisphere was dominant for processing the current lexical items, findings which parallel those for normal speakers. PMID- 11011884 TI - Response amendment in fencing: differences between elite and novice subjects. AB - Reaction time (RT), movement time (MT), total response time (RMT), and accuracy of 3 elite and 3 novice fencers were studied under a dual response paradigm requiring a full lunge. Electromyographic activity (EMG) from selected arm and leg muscles was used to compare response profiles of the two groups. Although the elite subjects had slower MTs, their faster RTs resulted in significantly shorter total response times. The EMG analysis showed that in comparison to the novice subjects, onset of muscle activity was significantly faster for the elite group in five of the six muscles studied. In addition, the elite subjects showed more coherent muscle synergies and more consistent patterns of muscle coordination. Although the requirement to change targets (signalled by the arrival of a second stimulus) led to slightly more target misses for the elite group, the overall frequency was low, which indicates that it did not pose difficulty for either group. The present findings show that measures of response timing and neuromuscular coordination differentiate skill level in the fencing lunge and draw attention to practical implications for skill assessment and training. PMID- 11011885 TI - Tactual Performance Test: internal consistency reliability of the memory and location scores. AB - Internal consistency reliabilities were computed for the Tactual Performance Test Memory and Location scores (N=602). After adjusting for unequal item difficulty, the reliabilities for Memory and Location were .69 and .79, respectively. PMID- 11011886 TI - Further evidence of associations of type a personality scores and driving-related attitudes and behaviors. AB - The present study examined the effects of Type A personality on specific self reported driving attitudes and behaviors when operating a motor vehicle. 102 undergraduate students completed the student version of the Jenkins Activity Survey (Form T) and several questionnaires asking participants about their driving history, driving attitudes, and driving behaviors. When the full range of Type A scores were examined, Type A personality was significantly related to more traffic accidents, greater frequency of breaking traffic laws, higher impatience when driving, more displays of aggression on the road, and engaging in more risky driving behaviors (rs<.17). When extreme Type A and Type B scores were compared, Type A drivers reported being involved in significantly more motor vehicle accidents and reported displaying more aggression on the road. Further research should examine actual behavioral data using more diverse samples to validate the results. PMID- 11011887 TI - A new conceptual model of asymmetry in motor performance for bidimensional fast oscillating movements in selected variants of performance. AB - Spatial characteristics and lateral differences between two upper extremities were investigated in unilateral graphical tasks involving fast oscillating movements in the vertical plane based on the model of restricted (less than 10 degrees) horizontal abduction adduction in the shoulder joint. The spatial locations of reversal points were used to identify two groups of motor performance: with big angles and gross vertical vectors (stretched accordion group), and small projectile angles with small vertical vectors (compressed accordion group). Both groups appeared in right and left arm performance. The former group had a strong pattern of distribution of big and small projectile angles which reflects a particular variant of execution with a significant difference between angles and intermittent big and small angles (BB). Two other variants of execution relating to specific angular patterns of performance were identified in the compressed accordion group: one (Bs) showed a big difference between big and small angles but without intermittance; the other (ss) had only small differences between magnitudes of angles. The Bs variant of execution was observed only in left-handed performance, whilst ss was typical of both extremities. The performances affiliated to the stretched accordion group with the BB variant of execution mostly operated with reciprocal cooperation between alterations of X and Y vectors for the right arm. Performance related to the same group with the Bs variant of execution used concurrent collaboration involving alteration of these vectors for the left arm. The compressed accordion group which deployed the ss variant of execution mostly displayed concurrent alteration of vectors irrespective of the side of performance. It is suggested that the spatial movement strategies might reflect several different schemes of motor control wherein coupling of oscillators controls vertical and horizontal movements. It is also proposed that specific subunits of the functional system of nervous elements responsible for the expression of spatial derivatives of motor programmes may exist at lower levels of the CNS and might be initiated by the left brain or by the cooperative activity of the left and right hemispheres. PMID- 11011888 TI - Arousal and mood factors in the "Mozart effect". AB - Some investigators of the "Mozart effect" have not controlled for the influence of differences in arousal or mood induced by treatment conditions. Studies by Rideout and colleagues reported differences in spatial reasoning after listening to a Mozart sonata compared against a relaxation instruction tape. The conditions may have affected subjects' arousal differentially, with the sonata increasing arousal and the relaxation instructions decreasing arousal, which could have affected spatial reasoning performance. Evidence is cited in support of this suggestion and indicates the importance of analyzing the influence of arousal differences in Mozart effect research. PMID- 11011889 TI - Re-evaluating Rudolf Laban's choreutics. AB - The choreutic conception of kinesthetic-motor space originally proposed by Rudolf Laban is given psychological validation through comparisons with similar conceptual models in spatial cognition and motor control research. Highly differentiated systems of reference are distinguished. Spatial paths are mentally represented as a series of locations linked into polygonal trajectories. Map-like images of the kinesphere' are structured with polyhedral networks. A fundamental practice consists of transforming spatial information with various operations (rotation, reflection, sizing, etc.). Loci are linked into spatial 'scales' which provide symmetrical solutions to geometric puzzles such as the traveling salesman problem and those devised by Hamilton (Icosian game) and Euler (Koenigsberg bridge problem). In the body these are considered to be prototypes for spatial paths used during adjustments of dynamic equilibrium. The choreutic conception of polyhedral body space is especially similar to the trajectory formation model. The choreutic principal of topological forms deflecting across various kinespheric nets is analogous to N. Bernstein's description of the "net of the motor field ... as oscillating like a cobweb in the wind." These commonalities indicate that further critical reviews of "choreutic laws" might be valuable in suggesting conceptual models and analytical tools for movement study in cognitive science. PMID- 11011890 TI - Indoor travel and simple tasks as physical exercise for people with profound multiple disabilities. AB - 2 to 4 30-min. periods of indoor travel and simple tasks were arranged daily as physical exercise for two women with profound mental retardation, deafness, and profound visual impairment. The questions were whether (a) the women could manage this fairly extensive exercise regimen (over about 9 and 4.5 mo.) with support technology and only minimal staff supervision and (b) the exercise could affect aspects of the women's physical condition. Data indicated that both women could successfully manage the exercise regimen with support technology, and the exercise seemed to have some beneficial effects on physical aspects such as bone metabolism and step width. PMID- 11011891 TI - Bisection of shapes and lines: analysis of the visual and motor aspects of pseudoneglect. AB - This study examined the visual and motor components of pseudoneglect as expressed on horizontal bisection tasks. Ten participants were tested on bisection of lines and elliptical shapes and judgement of pretransected lines. Results showed the same magnitude of leftward error on all three tasks, in contrast to previous findings for patients with visuospatial neglect who are known to bisect ellipses and circles much more accurately than lines. Participants' mean line bisection error was significantly further to the left than the mean subjective midpoint found on the judgement task. The findings contradict the claim that the visual component of pseudoneglect is caused by the same mechanism that causes visuospatial neglect but support the hypothesis that the motor component of pseudoneglect is similar to directional hypometria. PMID- 11011892 TI - Recognition of emotional facial expressions in depressed children and adolescents. AB - Of the neurobiological models of children's and adolescents' depression, the neuropsychological one is considered here. Experimental and clinical evidence has allowed us to identify a lateralization of emotional functions from the very beginning of development, and a right hemisphere dominance for emotions is by now well-known. Many studies have also correlated depression with a right hemisphere dysfunction in patients of different ages. The aim of our study was to analyze recognition of different facial emotions by a group of depressed children and adolescents. Patients affected by Major Depressive Disorder recognized less fear in six fundamental emotions than a group of healthy controls, and Dysthymic subjects recognized less anger. The group of patients' failure to recognize negative-aroused facial expressions could indicate a subtle right hemisphere dysfunction in depressed children and adolescents. PMID- 11011893 TI - Decision making in dual-task environments: analysis of hemispheric competition effects. AB - Performance degradations in multitasking situations have been reported frequently as a predictable effect of competition that arises from different processing demands whose hemispheric locations are too proximal. This model might be useful in explaining performance deficits in complex workplaces. To test this assertion, a laboratory study was designed to create an analogue of the processing demands required by a tactical decision-making task performed by 24 right-handed men. Vocalization, dichotic listening and decision-making performance were assessed under single- and dual-task conditions. The results were consistent with the predictions from hemispheric competition in the case of dichotic listening but not with vocalization. The results are discussed in terms of their implications for both research and systems design. PMID- 11011894 TI - Relationships between physical education classes and the enhancement of fifth grade pupils' self-concept. AB - Physical education classes given to a group of 46 pupils (27 boys and 19 girls) over a 6-mo. term differed from the usual program (control group of 29 pupils) in identifying individual goals, promoting participative and noncompetitive games, and praising students every chance possible during the classes. The enhancement of self-concept, particularly the physical self-concept, was significant for the experimental group as measured by the Self-description Questionnaire and the Child and Youth Physical Self-perception Profile. PMID- 11011895 TI - Differences in reactions to paintings by male and female college students. AB - 38 male and 55 female college students rated digitized color facsimiles of 40 paintings that varied in artistic period (Renaissance, Rococo, Impressionist, Post-Impressionist, 20th Century) and subject matter (portrait, landscape, still life, behavior depiction) on 12 7-point semantic differential scales, e.g., simple-complex. Women judged the content of Rococo and Impressionist paintings as more pleasing than did men and Impressionist paintings evoked greater feelings of pleasure and relaxation among women than among men. In addition, paintings that depicted behaviors evoked more pleasure and alertness among women than among men. The results were interpreted in terms of underlying differences between men and women in perceptual style and emotional sensitivity. PMID- 11011896 TI - Thoughts regarding the qualitative analysis of human movement. AB - The qualitative analysis of human movement is composed of many parts. Trying to assess the contribution of each of these components to the whole process of qualitative analysis is difficult. Evaluating a qualitative approach to human movement analysis using quantitative procedures can lead to misinterpretations. Qualitative analysis of movement has components important to its success which cannot be gauged by traditional biomechanical standards. PMID- 11011897 TI - Optimization characteristics of walking with and without a load on the trunk of the body. AB - Previous work showed that subjects naturally adopt a walking speed which optimizes energy cost of locomotion and stability of stride; however, no studies have examined whether these criteria are affected by carrying an external load. The purpose of this study was to compare optimization characteristics during loaded or unloaded walking. Energy cost and stride characteristics were measured for 10 subjects with and without a load on the trunk of the body of 10% of the body weight during 4 sessions. The first 2 sessions represent free walking at 2.5, 3, 3.5, 4, 4.5, and 5 km x hr.(-1). The last sessions represent free vs forced walking at constant speed (preferred frequency and +/- 10 PF and +/-20% of preferred frequency). Results show an effect of load on energy cost of walking but no effect on the optimal speed for stability. Furthermore, when carrying a load the subject does not adopt systematically the speed that minimizes physiological cost. Our findings suggest the necessity to consider this effect to prevent gait disturbance and maintain the health benefits of walking. PMID- 11011898 TI - Right to left hemispheric shift in occipital electroencephalographic responses to repeated Kimura figures. AB - Gordon and Carmon (1976) reported that repeated presentations of an initially novel stimulus were associated with a transfer of cerebral dominance over time (trials) from the right to the left hemisphere. To test the generalizability of these results the proportions of alpha rhythms over the left and right occipital and parietal lobes were measured following the presentation of recurring and nonrecurring complex visual patterns (the Kimura Figures) to the upper or lower, left or right peripheral visual fields. Analysis showed increased electrical activation (as inferred by attenuated proportions of alpha rhythms) of the left occipital lobe but decreased activation of the right occipital lobe. This shift occurred during repeated presentations of the same stimuli but not during single presentations of different novel stimuli. There was no significant shift in this activity over the parietal lobes. These results are consistent with the reports of other researchers who have found a shift of dominant neuroelectrical activity from the right to the left hemisphere as the novelty of a visuospatial stimulus decreases. PMID- 11011899 TI - Relation of useful field of view and other screening tests to on-road driving performance. AB - The purpose of this study was to examine the value of a clinical driving assessment battery in predicting performance on an on-road driving test. 43 participants referred to the Bryn Mawr Rehab Adapted Driving Program for evaluation of driving ability underwent an evaluation consisting of a predriver screening and an on road driving test. The predriver screening included a vision screening, a reaction rime task, a split-attention task, the Hooper Visual Organization Test, verbal and symbolic sign recognition, and assessment of Useful Field of View. Logistic regression analyses were applied to identify which predriver screening variables could be used to predict outcome on the on road driving test (pass/fail); UFOV was that best single predictor. The addition of screening tests beyond UFOV alone did not increase predictive validity. These findings suggest that UFOV may serve as an indicator of the need for further driving assessment. PMID- 11011900 TI - Chronic stress affects blood pressure and speed of short-term memory. AB - Subjects high and low on chronic stress (daily hassles) were engaged in a short term memory-scanning task with (n = 12) or without (n = 12) an active listening task. Blood pressure and heart rate were obtained before and during the experiment. Subjects with high stress were slower in short-term memory processing, had higher systolic blood pressure and heart rate (at baseline and during task performance), and reported more health complaints than those low in stress. The processing efficiency theory may predict slower memory scanning in subjects with high chronic stress, as part of these subjects' working memory capacity is taken up by more worries than that of the other subjects. Speed of short-term memory was even lower for subjects with high stress when performing the memory task in the active listening condition. PMID- 11011901 TI - Effects of speaker variability and phoneme location on speech perception. AB - The effects of variation in a speaker's voice and temporal phoneme location were assessed through a series of speeded classification experiments. Listeners monitored speech syllables for target consonants or vowels. The results showed that speaker variability and phoneme-location variability had detrimental effects on classification latencies for target sounds. In addition, an interaction between variables showed that the speaker variability effect was obtained only when temporal phoneme location was fixed across trials. A subadditive decrement in latencies produced by the interaction of the two variables was also obtained, suggesting that perceptual loads may not affect perceptual adjustments to a speaker's voice in the same way that memory loads do. PMID- 11011902 TI - Assessment of visual-motor coordination in 6- to 11-yr.-olds. AB - The concurrent and content validity of the Rey-Osterrieth Complex Figure Test and the Beery Developmental Test of Visual-Motor Integration-Third Revision were investigated through correlational analysis. 432 children, ages 6 to 11 years, were administered both tests. Across age groups participants performed better on both measures, providing support for the similarity of the measures and their sensitivity to the development of visuomotor integration. Although analysis indicated considerable overlap in the content of the two scales, the shared variance ranged from 7% to 31%, depending on the age of the child. The Rey Osterrieth figure is composed of overlapping squares, rectangles, triangles, and various other shapes. Given this complex combination, scores on this test reflect the examinees' visual organization and motor planning skills. On the other hand, the Beery test consists of a series of shapes which progress from simple figures to more complex ones. Because the figures become more difficult to copy, the score on this test reflects the examinees' developmental level of visuomotor ability. Despite these differences in test stimuli and interpretation of performance, the present study showed considerable shared variance in the scores of the examinees who took both tests. Finally, local norms for the Rey-Osterrieth figure using the scoring approach of E. M. Taylor (1959, adapted from Osterrieth, 1944) and including standard scores are presented for children ages 6 to 11 years. PMID- 11011903 TI - Effects of minimal exercise and cognitive behavior modification on adherence, emotion change, self-image, and physical change in obese women. AB - The present investigation tested a 12-wk. treatment protocol which employed low intensity cardiovascular and resistance exercise as well as cognitive-behavior modification on 13 obese, previously sedentary women. Separate analyses were conducted on program maintenance, emotional change, and physiological change. Although self motivation was lower in the treatment group than in the control group (n=35), measures of exercise maintenance were significantly higher. Analyses within the treatment group only indicated significant improvements in measures of State Anxiety, Depression, Anger, Fatigue, Tension, and Vigor, also Health Evaluation, Body Area Satisfaction, and heightened Overweight Preoccupation, over the 12 weeks. Their feelings after individual bouts of exercise indicated significantly increased Positive Engagement, Revitalization, and Tranquility, and reduced Physical Exhaustion. Maximum volume of oxygen uptake (VO2max) significantly increased (2nd to 10th percentile), but not resting heart rate. No significant correlation was found between cardiorespiratory change and change in scores on depression and anxiety. No significant association was found between physiological change and change in body image. Preliminary evaluation of the minimal exercise treatment was given. The need to replicate findings with larger and different samples was emphasized. PMID- 11011904 TI - Environmental events and induction of metamotivational reversals. AB - Three administrations of the Telic State Measure were used to test the notion from reversal theory that environmental events may induce reversals in metamotivational state. Significant changes in scores on Telic State Measure items from three different groups of students experiencing three different environmental events and one student control group (total N=96) confirmed that unexpected environmental events could induce reversals from the telic to the paratelic metamotivational state. PMID- 11011905 TI - Development of a random response scale for the multiple affect adjective check list-revised. AB - A scale was constructed to identify random responses on the Multiple Affect Adjective Check List-Revised. Items chosen were the 14 least frequently checked items and 14 most frequently checked items, plus the seven most frequently checked negative items and the seven least frequently checked positive items (total=42). The Random Response Scale successfully differentiated random protocols from those produced by 420 college students, and scores on the scale were significantly higher for the college students than for the random sample. In addition, correlations between scores on the Random Response Scale and the Communality Scale (Adjective Check List) and the NEO-FFI Conscientiousness Scale suggest its usefulness as a measure of "conscientiousness" or "dependableness." PMID- 11011906 TI - Illusory correlation: a function of availability or representativeness heuristics? AB - The present study sought to investigate the illusory correlation phenomenon by experimentally manipulating the availability of information through the use of the "lag" effect (Madigan, 1969). Seventy-four university students voluntarily participated in this study. Similar to Starr and Katkin's (1969) methodology, subjects were visually presented with each possible combination of four experimental problem descriptions and four sentence completions that were paired and shown twice at each of four lags (i.e., with 0, 2, 8 and 20 intervening variables). Subjects were required to make judgements concerning the frequency with which sentence completions and problem descriptions co-occurred. In agreement with previous research (Starr & Katkin, 1969), the illusory correlation effect was found for specific descriptions and sentence completions. Results also yielded a significant effect of lag for mean ratings between 0 and 2 lags; however, there was no reliable increase in judged co-occurrence at lags 8 and 20. Evidence failed to support the hypothesis that greater availability, through the experimental manipulation of lag, would result in increased frequency of co occurrence judgements. Findings indicate that, in the present study, the illusory correlation effect is probably due to a situational bias based on the representativeness heuristic. PMID- 11011907 TI - Effects of obesity and body fat distribution on lipids and lipoproteins in nondiabetic American Indians: The Strong Heart Study. AB - OBJECTIVES: To examine the relationship between obesity and lipoprotein profiles and compare the effects of total obesity and central adiposity on lipids/lipoproteins in American Indians. RESEARCH METHODS AND PROCEDURES: Participants were 773 nondiabetic American Indian women and 739 men aged 45 to 74 years participating in the Strong Heart Study. Total obesity was estimated using body mass index (BMI). Central obesity was measured as waist circumference. Lipoprotein measures included triglycerides, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, apolipoprotein AI (apoAI), and apolipoprotein B (apoB). Partial and canonical correlation analyses were used to examine the associations between obesity and lipids/ lipoproteins. RESULTS: Women were more obese than men in Arizona (median BMI 32.1 vs. 29.2 kg/m2) and South Dakota and North Dakota (28.3 vs. 28.0 kg/m2), but there was no sex difference in waist circumference. Men had higher apoB and lower apoAI levels than did women. In women, when adjusted for center, gender, and age, BMI was significantly related to HDL cholesterol (r = -0.24, p < 0.001). There was a significant but weak relation with apoAI (r = -0.14, p < 0.001). Waist circumference was positively related to triglycerides (r = 0.14, p < 0.001) and negatively related to HDL cholesterol (r = -0.23, p < 0.001) and apoAI (r = 0.13, p < 0.001). In men, BMI was positively correlated with triglycerides (r = 0.30, p < 0.001) and negatively correlated with HDL cholesterol (r = -0.35, p < 0.001) and apoAI (r = -0.23, p < 0.001). Triglycerides increased with waist circumference (r = 0.30, p < 0.001) and HDL cholesterol decreased with waist circumference (r = -0.36, p < 0.001). In both women and men there was an inverted U-shaped relationship between obesity and waist with LDL cholesterol and apoB. In canonical correlation analysis, waist circumference received a greater weight (0.86) than did BMI (0.17) in women. However, the canonical weights were similar for waist (0.46) and BMI (0.56) in men. Only HDL cholesterol (-1.02) carried greater weight in women, whereas in men, triglycerides (0.50), and HDL cholesterol (-0.64) carried a large amount of weight. All the correlation coefficients between BMI, waist circumference, and the first canonical variable of lipids/lipoproteins or between the individual lipid/lipoprotein variables and the first canonical variable of obesity were smaller in women than in men. Triglycerides and HDL cholesterol showed clinically meaningful changes with BMI and waist circumference in men. All lipid/lipoprotein changes in women in relation to BMI and waist circumference were minimal. DISCUSSION: The main lipoprotein abnormality related to obesity in American Indians was decreased HDL cholesterol, especially in men. Central adiposity was more associated with abnormal lipid/lipoprotein profiles than general obesity in women; both were equally important in men. PMID- 11011908 TI - Lifetime overweight status in relation to serial changes in body composition and risk factors for cardiovascular disease: The Fels Longitudinal Study. AB - OBJECTIVE: The aims were to determine if 1) individuals who became and maintained overweight during their entire lifetime differ from those who were never overweight in terms of annual changes in adiposity and concurrent changes in cardiovascular disease (CVD) risk factors; 2) the changes and their relationships to each other varied between these groups or by sex within the groups; and 3) alcohol usage, smoking habits, and level of physical activity differed between groups. RESEARCH METHODS AND PROCEDURES: Data from 16,315 examinations of 414 individuals were utilized to assess lifetime overweight (body mass index [BMI] > 25 kg/m2) status. A regressive analytic approach was used to determine the average annual changes for each individual over an adult serial interval ranging from 4 to 20 years. RESULTS: Men and women who have become and maintained overweight have higher blood pressure and a poorer lipid/lipoprotein risk profile than those who have never been overweight. There is an accelerated deterioration in the atherogenic profile of overweight men and women as indicated by annual changes in CVD risk factors about double that of their never-overweight counterparts. In women, increased risk is derived from increasing systolic and diastolic blood pressure, whereas in men the increased risk comes not only from increasing diastolic blood pressure but also cholesterol, triglycerides, and low density lipoprotein cholesterol levels and, to a lesser extent, decreasing high density lipoprotein cholesterol. DISCUSSION: The reduced physical activity observed in the overweight adults may be related to their accumulation of adipose tissue at a rate about double their never-overweight counterparts, and this may be driving the higher rate of increase of CVD risk factors in the overweight groups. PMID- 11011909 TI - Effects of sibutramine plus orlistat in obese women following 1 year of treatment by sibutramine alone: a placebo-controlled trial. AB - OBJECTIVE: This study assessed whether adding orlistat to sibutramine would induce further weight loss in patients who previously had lost weight while taking sibutramine alone. RESEARCH METHODS AND PROCEDURES: Patients were 34 women with a mean age of 44.1 +/- 10.4 years, weight of 89.4 +/- 13.8 kg, and body mass index (BMI) of 33.9 +/- 4.9 kg/m2 who had lost an average of 11.6 +/- 9.2% of initial weight during the prior 1 year of treatment by sibutramine combined with lifestyle modification. Patients were randomly assigned, in double-blind fashion, to sibutramine plus orlistat or sibutramine plus placebo. In addition to medication, participants were provided five brief lifestyle modification visits during the 16-week continuation trial. RESULTS: Mean body weight did not change significantly in either treatment condition during the 16 weeks. The addition of orlistat to sibutramine did not induce further weight loss as compared with treatment by sibutramine alone (mean changes = +0.1 +/- 4.1 kg vs. +0.5 +/- 2.1 kg, respectively). DISCUSSION: These results must be interpreted with caution because of the study's small sample size. The findings, however, suggest that the combination of sibutramine and orlistat is unlikely to have additive effects that will yield mean losses > or =15% of initial weight, as desired by many obese individuals. PMID- 11011910 TI - Does weight loss maintenance become easier over time? AB - OBJECTIVE: Studies of health-related behaviors, including weight loss, have shown that risk of relapse decreases over time, although reasons for this relationship are unclear. The purpose of this cross-sectional study was to determine if subjects who have maintained weight losses for varying periods of time report different strategies for weight loss maintenance or differences in the effort and pleasure associated with weight maintenance behaviors. RESEARCH METHODS AND PROCEDURES: Subjects were 758 women and 173 men who had maintained losses of at least 30 lb (mean = 60 lb) for 2 years or longer (mean = 6.8 +/- 7.0 years). Self administered questionnaires assessed subjects' use of weight maintenance strategies in the past year and their perceptions of the effort, attention, and pleasure associated with weight maintenance. RESULTS: Subjects who had maintained weight losses longer used fewer weight maintenance strategies and reported that less effort was required to diet and maintain weight and that less attention was required to maintain weight. The pleasure derived from exercise, low-fat eating, and maintaining weight was unrelated to duration of weight loss maintenance. DISCUSSION: As duration increases, a shift in the balance between the effort and pleasure of weight maintenance may occur. This shift may increase the likelihood of continued maintenance. PMID- 11011911 TI - Occupational status, cortisol secretory pattern, and visceral obesity in middle aged men. AB - OBJECTIVE: Despite several studies indicating that social gradients are predictive of cardiovascular mortality, the pathogenetic mechanisms remain incompletely understood. RESEARCH METHODS AND PROCEDURES: A population sample of 51-year-old men (N = 284) was divided into a socioeconomic gradient with manual laborers, civil servants, and university graduates. Anthropometric measurements were registered. Cortisol concentrations were measured in saliva, collected repeatedly during an ordinary working day, and a low-dose dexamethasone suppression test was performed. RESULTS: Lower socioeconomic status was associated with visceral obesity and higher cortisol values in relation to perceived stress. However, total cortisol secretion over the day of study was not elevated. The regulation of cortisol secretion showed less plasticity and dexamethasone inhibition was less efficient in the men in the lower socioeconomic status group than in those with a higher socioeconomic status. These are known consequences of long term stress. Longer duration in low socioeconomic conditions seemed to worsen these phenomena. DISCUSSION: It was concluded that a low socioeconomic status is associated with perturbed cortisol secretion, which is elevated in relation to perceived stress. When the hypothalamic-pituitary-adrenal axis is subjected to prolonged increases in cortisol elicited by chronic stress, the regulation of cortisol secretion is affected, indicating neuroendocrine dysregulations. These observations may provide a means for understanding the association of socioeconomic impairments with visceral obesity and the social inequality in risk for prevalent and serious diseases. PMID- 11011912 TI - Teasing history, onset of obesity, current eating disorder psychopathology, body dissatisfaction, and psychological functioning in binge eating disorder. AB - OBJECTIVE: The primary goal of this study was to examine associations among teasing history, onset of obesity, current eating disorder psychopathology, body dissatisfaction, and psychological functioning in women with Binge Eating Disorder (BED). RESEARCH METHODS AND PROCEDURES: Subjects were 115 female adults who met DSM-IV criteria for BED. Measurements assessing teasing history (general appearance [GAT] and weight and size [WST] teasing), current eating disorder psychopathology (binge frequency, eating restraint, and concerns regarding eating, shape, and weight), body dissatisfaction, and psychological functioning (depression and self-esteem) were obtained. RESULTS: History of GAT, but not WST, was associated with current weight concerns and body dissatisfaction, whereas both GAT and WST were significantly associated with current psychological functioning. Patients with earlier onset of obesity reported more WST than patients with later onset of obesity, but the groups did not differ significantly in GAT, current eating disorder psychopathology, body dissatisfaction. or psychological functioning. Obese women reported more WST than non-obese women, but no differences in GAT or the other outcome variables were observed. Higher frequency of GAT was associated with greater binge frequency in obese women, and with greater eating restraint in non-obese women. DISCUSSION: Although physical appearance teasing history is not associated with variability in most eating disorder psychopathology, it is associated with related functioning, most notably body dissatisfaction, depression, and self-esteem. Our findings also suggest that the age of onset of obesity and current body mass index status in isolation are not associated with eating psychopathology or associated psychological functioning in adult patients with BED. PMID- 11011913 TI - Obesity management: Australian general practitioners' attitudes and practices. AB - OBJECTIVES: To document general practitioners' (GPs) attitudes and practices regarding the prevention and management of overweight and obesity. RESEARCH METHODS AND PROCEDURES: A cross-sectional survey of a randomly selected sample of 1500 Australian GPs was conducted, of which 752 questionnaires were returned. The measures included views on weight management, definitions of success, views regarding the usefulness of drugs, approaches to and strategies recommended for weight management, and problems and frustrations in managing overweight and obesity. RESULTS: GPs view weight management as important and feel they have an important role to play. Although they consider themselves to be well prepared to treat overweight patients, they believe that they have limited efficacy in weight management and find it professionally unrewarding. GPs view the assessment of a patient's dietary and physical activity habits and the provision of dietary and physical activity advice as very important. The approaches least likely to be considered important and/or least likely to be practiced were those that would support the patient in achieving and maintaining lifestyle change. DISCUSSION: There remains considerable opportunity to improve the practice of GPs in their management of overweight and obesity. Although education is fundamental, it is important to acknowledge the constraints of the GPs' existing working environment. PMID- 11011914 TI - Differential expression of leptin receptor in high- and low-fat-fed Osborne Mendel and S5B/Pl rats. AB - OBJECTIVE: The regulation of body weight and body composition involves input from genes and the environment. This interaction is demonstrated by the different susceptibility of Osborne-Mendel (OM) and S5B/P1 rat strains to obesity when offered a high-fat diet. In animals and humans, diet-induced obesity has been characterized by hyperleptinemia, which has been interpreted as evidence for leptin resistance. This investigation determined if altered expression of leptin receptors (ObR) in the hypothalamus could potentially contribute to altered sensitivity to diet-induced obesity between OM and S5B/P1 rats. RESEARCH METHODS AND PROCEDURES: OM and S5B/P1 rats were fed high-fat (HF) or low-fat (LF) diets for 14 days. Ribonuclease protection assays and Western blotting were used to assay the levels of mRNA and protein, respectively, for short (ObR-S) and long (ObR-L) forms of the leptin receptor in the hypothalamus. RESULTS: The mRNA encoding ObR-L, the predominant signaling form of the receptor, was higher in OM rats than in S5B/P1 rats (p < 0.01) both on HF and LF diets. No changes in ObR-L mRNA expression were observed in OM rats with diet, but, S5B/P1 rats showed a slight increase in the ObR-L on the LF diet. On the contrary, there were no changes in ObR-S mRNA expression due to diet or strain. Western blots showed that both the short and long forms of the receptor were increased on the LF diet, but there were no strain differences. OM and SSB/P1 rats had comparable leptin levels after maintenance on a LF diet (6.20 +/- 0.63 and 4.81 +/- 0.82 ng/mL, respectively). Serum leptin levels in OM rats were increased by the HF diet and were elevated 2-fold over those of their S5B/P1 counterparts. DISCUSSION: These results suggest that a decrease in the levels of both the long form and short form of the receptor may contribute to the leptin resistance seen in HF-fed rats. These effects appear to be post-transcriptional, because equivalent changes were not observed in the expression of ObR-L and ObR-S mRNAs. They may be related to the increase in circulating leptin levels, suggesting that high serum leptin levels contribute to increased leptin resistance and subsequently lead to obesity. We conclude that down-regulation of receptor protein levels is associated with hypothalamic leptin resistance of HF-fed rats. PMID- 11011915 TI - Augmented insulinotropic action of arachidonic acid through the lipoxygenase pathway in the obese Zucker rat. AB - OBJECTIVE: The metabolism of arachidonic acid (AA) has been shown to be altered in severe insulin resistance that is present in obese (fa/fa) Zucker rats. We examined the effects and mechanism of action of AA on basal and glucose stimulated insulin secretion in pancreatic islets isolated from obese (fa/fa) Zucker rats and their homozygous lean (Fa/Fa) littermates. RESEARCH METHODS AND PROCEDURES: Islets were isolated from 10- to 12-week-old rats and incubated for 45 minutes in glucose concentrations ranging from 3.3 to 16.7 mM with or without inhibitors of the cyclooxygenase or lipoxygenase pathways. Medium insulin concentrations were measured by radioimmunoassay, and islet production of the 12 lipoxygenase metabolite, 12-hydroxyeicosatetraenoic acid (12-HETE), was measured by enzyme immunoassay. RESULTS: In islets from lean animals, AA stimulated insulin secretion at submaximally stimulatory glucose levels (<11.1 mM) but not at 16.7 mM glucose. In contrast, in islets derived from obese rats, AA potentiated insulin secretion at all glucose concentrations. AA-induced insulin secretion was augmented in islets from obese compared with lean rats at high concentrations of AA in the presence of 3.3 mM glucose. Furthermore, the inhibitor of 12-lipoxygenase, esculetin (0.5 microM), inhibited AA-stimulated insulin secretion in islets from obese but not lean rats. Finally, the islet production of the 12-HETE was markedly enhanced in islets from obese rats, both in response to 16.7 mM glucose and to AA. DISCUSSION: The insulin secretory response to AA is augmented in islets from obese Zucker rats by a mechanism related to enhanced activity of the 12-lipoxygenase pathway. Therefore, augmented action of AA may be a mechanism underlying the adaptation of insulin secretion to the increased demand caused by insulin resistance in these animals. PMID- 11011916 TI - Allergic contact dermatitis from hydrangea--is it so rare? AB - 8 cases of allergic contact dermatitis from hydrangea seen in Angers, France, during the last 15 years are reported and compared to other cases found in the literature. In this review, allergic contact dermatitis from hydrangeas appears to be an occupational dermatosis among nursery workers, presenting as an eczema involving the hands and especially the first 3 fingers, with chronic features of fissuring and scaling and with a chronic course. Differential diagnosis from irritant contact dermatitis may be difficult. Patch tests with the stem as well as the leaf of hydrangeas gave strong positive reactions in all patients, and hydrangenol, the allergen of hydrangea, when tested, always also gave a positive reaction. Sensitization seems to occur after close and prolonged contact with the plant, which could explain the relative frequency in Angers because it provides almost 90% of hydrangea seedling production in France. PMID- 11011917 TI - Contact allergy and hand eczema in Swedish dentists. AB - Hand eczema and contact allergy in Swedish dentists were studied in a multidisciplinary project. The aims of the study were to establish diagnoses, to investigate the occurrence of contact allergy, in particular to (meth)acrylates, and to evaluate certain consequences of hand eczema. A postal questionnaire on skin symptoms, atopy and occupational experience was mailed to 3,500 dentists aged <65 years, and licensed 1965-1995. The response rate was 88%. Among dentists living in 3 major cities, 14.9% (n= 191) reported hand eczema during the previous year. They were invited to a clinical examination, including patch testing with a standard and a dental series. 158/191 (83%) dentists attended, and hand eczema diagnosis was confirmed in 149/158 (94%). Irritant contact dermatitis was diagnosed in 67% and allergic contact dermatitis in 28%. On patch testing, 50% presented at least 1 positive reaction. The most frequent allergens were nickel sulfate, fragrance mix, gold sodium thiosulfate and thiuram mix. 7 (5%) had positive reactions to (meth)acrylates, all to 2-hydroxyethyl methacrylate and 6 also to ethyleneglycol dimethacrylate. 38% had consulted a physician, 4% had been on sick-leave and 1% had changed occupational tasks due to hand eczema. No dentist with allergy to acrylates had been on sick-leave or changed occupation. It is concluded that dentistry is a high-risk occupation for hand eczema, and that irritant contact dermatitis is most common. The prevalence of contact allergy to acrylates was below 1% in the population of responding dentists, and in most cases did not have serious medical, social or occupational consequences. PMID- 11011918 TI - Dyshidrotic eczema and occupation: a descriptive study. AB - Dyshidrotic eczema (DE) is a chronic and/or recurrent vesicular eczema of the palmar sides of the hands and sometimes the soles of the feet. The major aetiologies to be considered are atopic DE, contact DE, systemic contact DE and DE due to fungal or microbial allergy. To clarify the role of occupational contact allergens, 59 workers with DE were examined to determine whether they had atopy, environmental contact sensitization and/or contact allergy coupled with work activity. For all, patch testing was carried out using the European standard series and the material brought in by the patients. No significant difference was found between the rate (42.4%) of history of atopy among the 59 workers with DE and that (30.6%) observed among 160 workers with other forms of hand eczema (HE). Positive patch test reactions were found in 72.8% of dyshidrotic patients. A relationship to occupational activity was strong for 18 (30.5%) of them but this rate was significantly lower than that (55%) observed among the 160 with other types of eczema. Results of the present study showed that contact sensitization is as much involved in workers with DE as in workers with other form of HE, even though the relationship to work is significantly less frequent for DE. It also suggests that atopic histories are no more involved in DE than in other forms of HE. PMID- 11011919 TI - Lymphocyte subset changes in blood and gastrointestinal mucosa after oral nickel challenge in nickel-sensitized women. AB - This study investigates lymphocyte subsets in both the gastrointestinal mucosa and blood, in patients with nickel allergic contact dermatitis, after 10 mg oral nickel challenge (double-blind, placebo-controlled). 6 such patients with cutaneous symptoms induced only by skin contact with nickel (group A), 6 with a flare-up of cutaneous symptoms after food nickel ingestion (group B) and 6 healthy controls (group C) were enrolled. Blood lymphocyte subsets (CD4, CD45RO, CD8) were analyzed before and after 4 and 24 h from the challenge (test 1, 2, and 3), and intestinal biopsies were performed 2 days later. Challenges were positive in group B and negative in group A and controls. Serum and urine nickel levels significantly increased after nickel ingestion, with no differences between the 3 groups. At test 3, a significant decrease of the all CDs studied was found in group B. Biopsies of this group showed higher levels of CD45RO+ cells in the lamina propria and in the epithelium and lower levels of epithelial CD8+ lymphocytes. This study confirms that ingested nickel may induce flare-up of cutaneous reactions in some nickel-allergic patients, independently of the degree of sensitization and the intake of metal. In these patients, oral nickel stimulates the immune system, inducing maturation of T lymphocytes from virgin into memory cells; these latter cells seem to accumulate in the intestinal mucosa. The immunoreaction also involves CD8+ cells, whose role is not yet clear. PMID- 11011920 TI - Repeated exposures to cobalt or chromate on the hands of patients with hand eczema and contact allergy to that metal. AB - The present study aimed at evaluating the effects of daily repeated exposures to low cobalt or chromate concentrations on the hands of patients with hand eczema and cobalt or chromate allergy. For 2 weeks, the patients immersed a finger for 10 min daily into the appropriate metal salt solution in water. During the 1st week, this was a 10 or 50 mg/l cobalt concentration or a 10 mg/l chromate concentration, and, during the 2nd week, a 100 or 200 mg/l cobalt concentration or a 100 mg/l chromate concentration. This regimen elicited a flare of hand eczema only in the chromate-exposed chromate-sensitive patients. During the exposure period, accumulation of cobalt or chromate in the nail was demonstrated. Standardization of chemical methods of quantification of skin exposure to allergens, combined with experimental exposure studies in patients with specific contact allergy, will increase the possibility of providing evidence-based medicine in the area of allergic contact dermatitis in the future. PMID- 11011922 TI - Contact urticaria from panthenol in hair conditioner. PMID- 11011921 TI - Multicentre study of allergic contact cheilitis from toothpastes. AB - The present work reports the results of a multicentre study of toothpaste allergic contact cheilitis (TACC) conducted by GIRDCA (Gruppo Italiano Ricerca Dermatiti da Contatto e Ambientali). The study examined 54 patients with eczematous lesions on the lips, the possible cause of which was suspected to be the use of toothpastes. Patch tests were conducted with a standard series, a specially-targeted series (toothpaste cheilitis series, TCS), and with suspected toothpaste(s). A stop-restart test (SRT) was carried out with these, together with a use test to identify possible alternative products. The TCS produced 17 positive reactions in 13 patients, the most frequent being to spearmint oil. Of the 54 patients, 5 displayed positive reactions only to the TCS. The patch tests with toothpaste produced positive reactions in 11/32 patients, the SRT a positive response in 10/12 cases. The diagnosis of TACC was confirmed in 15/54 patients. Alternative products were identified for 5 patients. In conclusion, the allergens most frequently responsible for TACC were the flavourings, and the additional series proved to be useful in many cases (together with patch tests with toothpastes and the SRT) for correct diagnosis and to initiate effective prevention. PMID- 11011923 TI - Allergic contact dermatitis due to diphenylthiourea in a neoprene slimming suit. PMID- 11011924 TI - Occupational contact dermatitis from diethylene glycol monobutyl ether in a podiatrist. PMID- 11011925 TI - A case of contact hypersensitivity to platinum salts. PMID- 11011926 TI - A series of 3 patients sensitized to reactive dyes during patch testing. PMID- 11011927 TI - The role of nickel in oral disease. PMID- 11011928 TI - Photoallergic eczema caused by sunscreens in a 12-year-old girl. PMID- 11011929 TI - Occupational contact urticaria syndrome from cefotiam dihydrochloride in a latex allergic nurse. PMID- 11011930 TI - Contact allergy to doxepin. PMID- 11011931 TI - Chronic urticaria from an acrylic dental prosthesis. PMID- 11011933 TI - Allergic contact dermatitis due to 1,3-butylene glycol. PMID- 11011932 TI - Allergic contact dermatitis from diethyl sebacate in lanoconazole cream. PMID- 11011934 TI - An outbreak of occupational textile dye dermatitis from Disperse Blue 106. PMID- 11011935 TI - Di(2-ethylhexyl) phthalate (DOP) in the dotted polyvinyl-chloride grip of cotton gloves as a cause of contact urticaria syndrome. PMID- 11011937 TI - Cucumber contact dermatitis. PMID- 11011936 TI - The biocide iodopropynyl butylcarbamate (IPBC) as an allergen in cutting oils. PMID- 11011938 TI - Allergic contact dermatitis from formaldehyde and quaternium-15 in photocopier toner. PMID- 11011940 TI - Allergic contact dermatitis from chromate and 4-tert-butylphenol-formaldehyde resin in a father and daughter. PMID- 11011939 TI - Allergic contact dermatitis from airborne exposure to acrylates. PMID- 11011941 TI - Hydrolyzed protein shampoo additives are not a common contact allergen. PMID- 11011942 TI - Contact anaphylaxis due to hair dye. PMID- 11011943 TI - Immediate, anaphylactic and delayed reactions to jellyfish. PMID- 11011944 TI - Occupational UVA-induced allergic photodermatitis in a welder due to hydrochlorothiazide and ramipril. PMID- 11011945 TI - Systemic contact dermatitis from implanted disulfiram. PMID- 11011946 TI - Sorbic acid: an unusual cause of systemic contact dermatitis in an infant. PMID- 11011947 TI - Contact dermatitis from parabens used as preservatives in eyedrops. PMID- 11011948 TI - Delayed-type hypersensitivity to heparin with tolerance of its intravenous administration. PMID- 11011949 TI - Occupational contact dermatitis from ylang-ylang oil. PMID- 11011950 TI - Overweight and obesity in European children and adolescents. AB - The conclusions of this mini-workshop are not substantially different from those previously published based on the USA experience [9]. This does not necessarily mean that there are no important differences between Europe and the USA, rather it reflects the paucity of data, especially in Europe. Moreover, European data are dominated by studies in Northern and Western European countries, which are closest in economic terms to the USA. This review has not been able to reflect the position in the less economically advantaged parts of Europe where more substantial differences may occur. Together, these reviews have highlighted the limited and inadequate data available on one of the most important and burgeoning public health problems in the world today. There is an urgent need for more research into the fundamental mechanisms of weight gain and effective interventions if we are to prevent the next generation becoming fatter than their parents. PMID- 11011951 TI - Overweight and obesity in European children and adolescents: causes and consequences, treatment and prevention. An introduction. PMID- 11011952 TI - Overweight and obesity in European children: definition and diagnostic procedures, risk factors and consequences for later health outcome. AB - In this article, information on the definition of obesity in childhood and adolescence, its differential diagnosis and its adverse health effects is provided. This information can be considered as a basis for the evaluation of an obese patient. Review of the international and European literature shows that the co-morbidities of childhood obesity cover a large medical area ranging from secondary hormonal disturbances to orthopaedic problems. Most of them have so far been underestimated. However, the most severe problem of an obese child is his or her psychosocial discrimination. Longitudinal data demonstrate a positive relationship between childhood obesity and increased morbidity and mortality in adulthood. CONCLUSION: Data from the literature are in some respect sparce since they do not allow to calculate the exact prevalence of adverse effects nor to assess the outcome of an obese child when it is successfully reducing the relative amount of body fat. PMID- 11011953 TI - Epidemiology of childhood obesity in Europe. AB - At present, estimation of the prevalence and secular trends in paediatric obesity in Europe is severely hampered by methodological problems in the definition of obesity and the paucity of data sets that mirror the demographic, cultural and socioeconomic composition of the European population. The available cross sectional data, however imperfect, suggest that there are complex patterns in prevalence which vary with time, age, sex and geographical region. Overall, the prevalence of obesity in young children is relatively low compared to adolescents. Gender differences in prevalence are inconsistent. The highest rates of obesity are observed in eastern and southern European countries and even within countries there may be marked variability in the rates of obesity. It is not clear whether the trends in paediatric obesity are a simple consequence of an overall increase in fatness in Europe or whether there may be sub-groups of children who, at certain ages, are either particularly susceptible to environmental challenges or are selectively exposed to such challenges. The respective contributions of dietary energy intake and patterns of physical activity to the aetiology of childhood obesity present a confused and confusing picture. Changing demographic and social circumstances throughout Europe are linked to childhood obesity but it is highly unlikely that these interact in similar ways in the genesis of obesity in different individuals and population groups. CONCLUSION: At present, our limited understanding of the variability in susceptibility to obesity in European children and adolescents provides powerful justification for the development of preventive strategies which are population based rather than selectively targeted at high-risk children. PMID- 11011954 TI - Aetiology of overweight and obesity in children and adolescents. AB - The epidemic diffusion of obesity in industrialised countries has promoted research on the aetiopathogenesis of this disorder. The purpose of this review is to focus mainly on the contribution that European research has made to this field. Available evidence suggests that obesity results from multiple interactions between genes and environment. Parents obesity is the most important risk factor for childhood obesity. Twin, adoption, and family studies indicated that inheritance is able to account for 25% to 40% of inter-individual difference in adiposity. Single gene defects leading to obesity have been discovered in animals and, in some cases, confirmed in humans as congenital leptin deficiency or congenital leptin receptor deficiency. However, in most cases, genes involved in weight gain do not directly cause obesity but they increase the susceptibility to fat gain in subjects exposed to a specific environment. Both genetic and environmental factors promote a positive energy balance which cause obesity. The relative inefficiency of self-adapting energy intake to energy requirements is responsible for fat gain in predisposed individuals. The role of the environment in the development of obesity is suggested by the rapid increase of the prevalence of obesity accompanying the rapid changes in the lifestyle of the population in the second half of this century. Early experiences with food, feeding practices and family food choices affect children's nutritional habits. In particular, the parents are responsible for food availability and accessibility in the home and they affect food preferences of their children. Diet composition, in particular fat intake, influences the development of obesity. The high energy density and palatability of fatty foods as well as their less satiating properties promotes food consumption. TV viewing, an inactivity and food intake promoter, was identified as a relevant risk factor for obesity in children. Sedentarity, i.e. a low physical activity level, is accompanied by a low fat oxidation rate in muscle and a low fat oxidation rate is a risk factor of fat gain or fat re-gain after weight loss. CONCLUSION: Further research is needed to identify new risk factors of childhood obesity, both in the genetic and environmental areas, which may help to develop more effective strategies for the prevention and treatment of obesity. PMID- 11011955 TI - Physical activity in relation to overweight and obesity in children and adolescents. AB - Fitness and physical activity levels of children and adolescents are commonly addressed but data on scientific investigations are both equivocal and methodologically diverse. The intensity and type of physical activity that benefit health and development during childhood are not known. Measurement of activity in children is problematic since there is no valid method of assessing activity levels that is feasible for use in large field studies. Most studies using self-report methods, heart rate studies using low heart rate threshold and doubly labelled water studies indicate relatively high levels of activity in children. The three national surveys on large representative samples reported that 60% to 70% of all children were involved in sufficient physical activity according to various definitions. Heart rate studies demonstrate that children generally perform short bouts of moderate to vigorous activities and seldom participate in long-sustained vigorous activities. They also proved that children perform large volumes of activity in the lower heart rate zones. It is generally accepted that boys are more active than girls and physical activity declines by age (peak around 13 to 14 years of age). The difference between the physical activity of European and North American children or between children living in different European countries is difficult to judge due to the diversity of methodology and definitions. CONCLUSION: There is a need to identify more clearly the quantity and type of activity which improves the health and promotes the normal development of children and to improve the methods assessing physical activity. PMID- 11011956 TI - Prevention and treatment of overweight and obesity in children and adolescents. AB - Increasing numbers of obese children and adolescents all over the world demand an investment in the primary and secondary prevention of obesity and overweight in this age group. The goal of preventive measures in children is to avoid the negative short- and long-term health problems associated with obesity. Primary prevention aims at establishing a healthy, active lifestyle and keeping children and adolescents within a range of body weight which is considered to be healthy. Constant availability and affordability of palatable and energy-dense food in the affluent society of the western world demands preventive strategies. Universal or public health prevention seems to be the most suitable form because several other cofactors of morbidity and mortality of affluent societies can also be prevented. However, in most European countries there is a lack of awareness of the necessity of prevention programmes, not only among the general population but also among the medical society. More awareness and consciousness to the problem of obesity must be generated in order to lead to effective therapeutic programmes. For those children and adolescents who are already obese, secondary prevention is mandatory. Therapeutic intervention programmes for the obese aim at long-term weight maintenance and normalisation of body weight and body fat. They have to modify eating and exercise behaviour of the obese child and establish new, healthier behaviour and lifestyle. Treatments programmes must include behavioural components in order to permanently change nutrition and physical exercise of the obese children and adolescents. However, long-term results of treatment programmes in European countries are scarce and the reported results, even of multidisciplinary regimens, are not impressive. CONCLUSION: In most European countries there is an urgent need not only for a growing awareness of the problem of obesity in children and adolescents but also for development of new comprehensive approaches in treating this group. PMID- 11011957 TI - Androgen receptor--an update of mechanisms of action in prostate cancer. AB - Androgen receptor (AR), a key nuclear transcription factor in the prostate gland, is expressed in all histological types and stages of prostate cancer. The AR regulates proliferation of prostate cancer cells by stimulation of cyclin dependent kinases. However, in some prostate tumors AR stimulates expression of cell cycle inhibitors, thus leading to down-regulation of cellular proliferation. Androgens, by activation of the AR, control differentiation of prostate cells and synthesis of neutral lipids. There are several mechanisms by which prostate cancer cells adapt to an environment with low androgen supply during endocrine therapy. The AR expression and activity increase in several cell lines that are used as an in vitro model for monitoring changes during long-term androgen ablation. Mutant ARs are of importance for monitoring the natural course of the disease and for determining the response to anti-androgens in metastatic lesions from prostatic carcinoma. In addition, AR activity is up-regulated by various stimulators of intracellular protein kinases. Current research efforts are focused on elucidation of function of AR coregulatory proteins, coactivators and corepressors. Their inappropriate expression and/or function might critically influence cellular events in advanced carcinoma of the prostate. It is hoped that information on these coregulatory proteins will serve as a basis for a more efficient pharmacological inhibition of the AR in advanced carcinoma of the prostate. PMID- 11011958 TI - Renal tubular apoptosis after complete ureteral obstruction in the presence of hyperoxaluria. AB - Hyperoxaluria is a well-known cause of renal stone disease and in vitro studies have shown that oxalate crystals have a stimulatory effect on apoptosis of renal tubular epithelial cells. Total and partial ureteral obstruction also have an accelerating effect on apoptosis of renal tubular epithelial cells. The aim of the present study was to investigate the apoptotic effect of unilateral ureteral obstruction in the presence of hyperoxaluria on the rat kidney. Twenty-eight male Wistar rats were divided into four groups, with seven rats in each. The groups were named G1 (control), G2 (hyperoxaluric), G3 (obstructive) and G4 (hyperoxaluric + obstructive). G2 and G4 rats were given 1% ethylene glycol (a precursor for oxalates) in their drinking water. G1 and G2 rats underwent sham operation, while left proximal ureteral ligation with a 5-zero silk suture was performed on G3 and G4 animals. The rats were sacrificed 2 weeks after the operation; left nephrectomy was then performed. We searched for the apoptotic cells by direct immuno-peroxidase detection of digoxigenin-labeled genomic DNA. The mean +/- SD values of the apoptotic cell count was 0.86+/-0.90 in G1 and 4.33+/-3.81 in G2. The values for G3 and G4 were 30.17+/-16.85 and 302.67+/ 184.45, respectively. We found a statistically significant difference between all groups (P < 0.001). When compared with the control group (G1), the mean apoptotic cell count was fivefold that of G2 and 35- and 351-fold those of G3 and G4, respectively. Our study demonstrated that hyperoxaluria with complete ureteral obstruction induces an excessive level of apoptosis, which is responsible for renal damage, and that ureteral obstruction is a more important factor for apoptosis than hyperoxaluria. Considering these data, we also believe that research studies for medical preventive measures must be considered for patients with ureteral obstruction and/or hyperoxaluria. PMID- 11011959 TI - The role of estrogen receptor, androgen receptor and growth factors in diethylstilbestrol-induced programming of prostate differentiation. AB - Recently, others and we have demonstrated that prenatal exposure to an extremely low dose of diethylstilbestrol (DES) and other estrogenic compounds produces a significant effect on mouse prostate development in vivo and in vitro in the presence and absence of androgen. In this study, we investigated the mechanism by which DES produces this effect and determined the role of its estrogenic activity on the growth and branching, induced by DES in the 17-day-old fetal prostate in culture. Additionally, we investigated whether the androgen receptor (AR) plays a role and whether any of the growth factors, namely, EGF and IGF-1 which are known to modulate the estrogen receptor (ER) and androgen receptor (AR)-dependent process, mediate the DES-induced effects. Using the organ culture bioassay of prostate development, we demonstrate that DES enhanced the growth and branching of the prostate at both 0.1 and 0.5 pg/ml dosages, thus, confirming a previous report of ours. An anti-estrogen, ICI164,387 blocked both of the effect of DES, suggesting that both of these two effects are ER dependent. Anti-androgen, flutamide also blocked both branching and prostatic growth induced by DES, while cyproterone acetate blocked only the branching effect, suggesting a role for AR in the DES-induced effects. Depletion of EGF by anti-EGF antibody blocked the DES induced effects and this was reversed following EGF replacement in the organ culture system. Anti-IGF-1 antibody, on the other hand, only blocked the branching effect, but produced no effect on the prostatic growth, induced by DES. Estrogenic chemicals, bisphenol A and DES enhanced EGF-mRNA level of the cultured prostates. Taken together, it appears that DES-induced prostatic enlargement involves enhancement of ER-dependent EGF and IGF-1 synthesis, mediating prostatic enlargement and androgen action. PMID- 11011960 TI - Epidermal growth factor in urine from patients with bladder cancer. AB - Epidermal growth factor (EGF), a mitogenic polypeptide with a molecular weight of 6000, is excreted in human urine in nanomolar quantities. Recently, some reports showed that urothelial neoplasm was related to the concentration of EGF in urine. In this study, EGF concentration in urine was measured by enzyme-linked immunosorbent assay (ELISA) in 207 samples from 112 male patients (30-90 years old, median 66.2) who had previously been treated for bladder cancer. Then, we tried to clarify the significance of urinary EGF as a marker for recurrence of bladder cancer in comparison with urine cytology. The samples were collected on occasional follow up cystoscopy. Urine from nine age-adjusted males without urological disease was also measured to obtain normal control values. In 123 samples from patients without tumors, EGF concentrations in urine decreased with age. In 84 samples obtained from patients with recurrent tumor, EGF concentrations were significantly lower than those in 123 samples from patients without tumors (P < 0.001) Furthermore, EGF concentrations in longitudinal samples collected the same patients during tumor recurrence and at the times when no tumor was detected were measured in 56 patients. EGF concentrations in the samples collected during tumor recurrence was significantly lower than that in specimens collected when there was no tumor (P < 0.01). There were no significant differences between the same samples collected during tumor recurrence with regard to tumor grade, stage shape and number of tumors. However, EGF concentration in urine from patients with carcinoma in situ (CIS) was lower than that in specimens from patients without CIS. These results indicate the usefulness of determining the EGF concentration as a marker for detecting bladder cancer recurrence. Urine cytology was also examined in the same series and findings were compared with those of urinary EGF. On cytology, class IV and V were considered positive, and on urinary EGF, less than 10 ng/mgCr were considered positive. Sensitivity was 25% for cytology and 57% for urinary EGF, while specificity was 98% and 66%, respectively. The predictive positive value was 0.88 and 0.53, respectively. With the combined use of urinary EGF and cytology, the sensitivity, specificity and predictive positive value were 68%, 64% and 0.92, respectively. In conclusion, urinary EGF seems to be a useful marker for detecting bladder cancer recurrence if performed in addition to cytology. cancer recurrence if performed in addition to cytology. PMID- 11011961 TI - Inhibition by vitamin C of DNA adduct formation and arylamine N-acetyltransferase activity in human bladder tumor cells. AB - Previous studies have already demonstrated the protective role of vitamin C (ascorbic acid) in certain types of cancer. This study reports on the effects of vitamin C on arylamine N-acetyltransferase (NAT) activity and DNA adduct formation in a human bladder tumor cell (T24) line. The activity of NAT was measured using high-performance liquid chromatography (HPLC), by assaying for the amounts of acetylated 2-aminofluorene (AF) and p-aminobenzoic acid (PABA) and the remaining amounts of AF and PABA. T24 cells were used for examining NAT activity and carcinogen DNA adduct formation. The results demonstrated that NAT activity and 2-aminofluorene DNA adduct formation in T24 cells were inhibited and decreased by vitamin C in a dose-dependent manner. The apparent kinetic parameters (apparent values of Km and Vmax) from T24 cells were also determined with and without vitamin C cotreatment. The data also indicated that vitamin C decreased the apparent values of Km and Vmax from T24 cells. PMID- 11011962 TI - Epithelial mucin expression in bladder cancer: correlation with pathological and clinical parameters. AB - Recently, attention has been drawn to the role of polymorphic epithelial mucin (PEM) as a possible target for cancer immunotherapy. To investigate the expression of this molecule in bladder tissue, we used two mouse monoclonal antibodies (HMFGI and HMFG2) raised against the core protein of the PEM. The localization of these two anti-PEM antibodies was examined in normal (n = 10), inflammatory (n = 10) and malignant (n = 67) bladder tissue samples with the use of a three-step avidin-biotin method. For HMFG1 and HMFG2 localization was successful in 78% and 60% of the bladder cancer samples, respectively, where as they were localized only in 30% and 40% of normal bladder tissue samples, respectively. Staining of either antibodies did not correlate with the grade, stage, or survival of bladder cancer patients. We conclude that PEM is frequently overexpressed by bladder cancer cells and HMFG1 is the antibody of choice to be used as a carrier of a cytotoxic agent for application of intravesical targeted therapy of bladder cancer. PMID- 11011963 TI - First experience with gamma probe guided sentinel lymph node surgery in penile cancer. AB - Because of the curative approach, the detection of lymph node metastases in squamous cell carcinoma (SCC) of the penis is of significant clinical relevance. Sentinel lymph node (SLN) identification by means of lymphangiography has been proven to be insufficiently safe. However, the high morbidity of inguinal lymphadenectomy and the considerable individual variability regarding the location of lymph node metastases justify the necessity of a technique that enables the identification of SLNs. Since 1998, SLNs have been intraoperatively identified and selectively dissected, after peritumoral injection of technetium 99m nanocolloid and using lymphoscintigraphy, in three patients (one with malignant melanoma and two with SCC). At least one SLN could be detected in each patient. The maximum surgical time was 30 min. There were no severe complications. Lymph node metastases did not occur in any patient. Upon a mean follow-up of 10 months, all patients are currently free of tumor. Owing to the long-term results of sentinel lymphadenectomy in malignant melanoma of other locations and our preliminary results with respect to penile carcinoma. we consider the current method appropriate as the only primary operation for lymph node staging in early stages and, in combination with modified inguinal lymphadenectomy, in locally advanced stages. PMID- 11011964 TI - An in vitro model for videoimaging of human bladder smooth muscle cell contractions. AB - Knowledge regarding human bladder smooth muscle cell (SMC) physiology is very limited. Only a few specific medical therapies for bladder disorders have therefore been established. The objective of this study was to develop a model for videomicroscopy of bladder SMC contractions. Cells were isolated from human cystoprostatectomy specimens and cultured in a modified EMEM medium. These cells were identified as SMCs by means of immunohistochemistry. For videomicroscopy, the culture flasks were coated with a viscous agent to allow cell contraction. Contractions were visualized by means of a cell culture microscope with a time lapse videosystem. For cholinergic stimulation of the cells, acetylcholine, in concentrations ranging from 100 microM to 10 mM, was applied. The percentage of contracting cells within the observation field was evaluated for quantitative analysis. In control experiments without contractile stimulant 6% of the cells were observed to contract. Stimulation with acetylcholine induced a significant dose-dependent increase to 47% in contracting cells. These results demonstrated that videomicroscopy is an appropriate tool to investigate the contraction mechanisms of bladder SMCs. This model offers the possibility of studying drug effects on the human detrusor in vitro. PMID- 11011966 TI - The role of intracellular and extracellular calcium in mechanical and intracellular electrical activity of human urinary bladder smooth muscle. AB - We studied the role of extracellular and intracellular Ca2+ in human detrusor smooth muscle contraction. Simultaneous recordings of mechanical and intracellular electrical activity were made in three different Ca2+ concentrations: normal Krebs' solution (100%), 10% of the standard Ca2+ concentration and a solution in which Ca2+ was omitted from the medium (0%). Spontaneous contractions and KCl or CCh induced contractions were studied. Ryanodine and caffeine were used to manipulate the intracellular Ca2+ stores. The present results show that only a very small amount of Ca2+ in the extracellular space is sufficient to support spontaneous and induced contractions. Spike-shaped potentials and long lasting depolarisations were recorded in all three solutions. However, the prevalence of long lasting depolarisations increased when the extracellular Ca2+ concentration was reduced. The amplitude of the spike-shaped potentials and long lasting depolarisations appeared to be negatively affected by diminishing the extracellular Ca2+ concentration. Additionally, the duration of the long lasting depolarisations was reduced in 0% Ca2+. The contraction upon KCl stimulation was primarily depending on the extracellular Ca2+. Upon muscarinic receptor stimulation, a combined activation of Ca2+ mobilisation from intracellular and extracellular stores may occur; the ratio of contribution of these two sources changes in accordance with the requirements of the conditions. PMID- 11011965 TI - Inhibition of diabetic bladder smooth muscle cell proliferation by endothelin receptor antagonists. AB - Urinary bladder hypertrophy and hyperplasia are well recognised in diabetic cystopathy. The urinary bladder is known to synthesise endothelin-1 (ET-1), a potent vasoconstrictor peptide with mitogenic properties. Using diabetic New Zealand White (NZW) rabbits, we investigated the potential role of ET receptor subtypes (ETA and ET(B)) on the proliferation of bladder smooth muscle cells (SMC). Diabetes mellitus was induced in adult male NZW rabbits. After 6 months, control (n = 6) and diabetic (n = 6) bladders were removed and SMC from the dome and bladder neck were grown using standard explant methodology. At passage two, the cells were made quiescent and then further incubated in foetal calf serum (FCS), control age-matched rabbit serum (CRS) or diabetic rabbit serum (DRS) in the presence or absence of ET(A)-antagonist (BQ123) or ET(B)-antagonist (BQ788). SMC proliferation was then measured with 5-bromo-2'deoxy-uracil 24 h later and by cell counting (using a haemocytometer) at 48 h. Neither BQ123 nor BQ788 influenced detrusor or bladder neck SMC proliferation in FCS or CRS. However, in the presence of DRS, BQ123 and BQ788 significantly inhibited diabetic detrusor and bladder neck SMC proliferation at 30 and 100 nmol/l (P < 0.03 and P < 0.01, respectively). Cell counts were also significantly reduced from the diabetic detrusor and bladder neck (P < 0.01 and P < 0.03 with BQ123 and BQ788, respectively). These results suggest that ET may play a pathophysiological role in the bladder SMC hyperplasia associated with diabetes mellitus. PMID- 11011967 TI - Determination of mouse bladder inflammatory response to E. coli lipopolysaccharide. AB - Evaluation of the severity of histologic changes associated with cystitis is often subjective and inconsistent from one sample to the next. The objective of this study was to establish a consistent, reproducible method to quantify histologic changes in a mouse model of lipopolysaccharide (LPS)-induced cystitis. Either LPS (n = 8) or pyrogen-free saline (n = 8) was instilled intravesically into the bladders of female C57bk-6 J mice. Twenty-four hours later, mice in these groups as well as eight untreated controls were sacrificed and bladders were removed, fixed in formalin, and stained with hematoxylin and eosin (H&E). A bladder inflammatory index (BII) was described by reviewing tissues for edema, leukocyte infiltration, and hemorrhage. Cross-sections were evaluated by a single pathologist in a blinded manner based on the objective BII described. The BII method for objectively analyzing bladder inflammation was effective and reproducible. Bladders instilled with LPS had significantly increased inflammation scores for edema, leukocyte infiltration, and hemorrhage compared with those instilled with saline or untreated controls (n = 8, P < 0.05). These results demonstrate that LPS causes bladder inflammation when instilled intravesically and that inflammation of mouse bladders can be objectively quantified using the histological method described. PMID- 11011968 TI - A new approach to calculate the risk of calcium oxalate crystallization from unprepared native urine. AB - This work focuses on the in vitro calcium-oxalate (CaOx) crystallization behaviour of native and synthetic urine samples in order to establish a CaOx crystallization risk index for unprepared native urine. Native 24-h urine samples from healthy persons and from stone-formers were examined. Within a [Ca2+] versus added oxalate (Ox2-) diagram, we observed fields which allow the discrimination of each urine sample in terms of more or less risk. The [Ca2+]/(Ox2-) ratio is calculated and termed the "Bonn-Risk Index" (BRI; per litre). We propose that BRIs > 1/l denote samples "at risk", whereas BRIs < or = 1/l denote those "without risk". Second. the effects of different concentrations of citrate and Mg2+ on BRI were investigated in artificial urine. The transferability of BRI between native and synthetic urine samples is proved. To evaluate the impact of the proposed BRI, it is compared with the more familiar relative urine saturation index calculated for CaOx and brushite. Urine sampled from stone-formers shows risk indexes between 0.278 and 23.0/l (mean 2.87/l), while urine from healthy persons varied between 0.060 and 4.890/l (mean 1.05/l). Comparing the results of healthy volunteers and patients, the significance of BRI and relative urine supersaturation (RS) with respect to CaOx is P < 0.0005 and P = 0.013, respectively. Fast and easy to perform, determination of the risk index is a suitable tool for estimating the actual CaOx formation "status"--"at risk" or "without risk"--from the native urine of any person. PMID- 11011969 TI - Poor semen quality from patients with malignancies does not rule out sperm banking. AB - Cancer therapy can further impair the already poor semen quality in cancer patients. This study evaluated the prefreeze and postthaw semen quality before treatment of patients with malignancies to examine the rationale for sperm banking for these men. Records of nine patients with different malignant tumors, who had been referred for sperm cryopreservation between 1982 and 1997, were reviewed and the results were compared with those of 50 normal healthy donors. Patients did not differ from donors in age, ejaculate volume, or duration of sexual abstinence. The total motile sperm count (median and interquartile range) was significantly different between patients and donors for prefreeze specimens (P = 0.026) and postthaw specimens (P = 0.008). Also, the percent motility was significantly lower in the patients as compared with the donors in prefreeze (P = 0.035) and postthaw specimens (P = 0.005). The percentage change in motility after thawing was also larger for patient samples (-54% versus -47%, P = 0.39). Other sperm motion characteristics did not significantly differ between the two groups except for postthaw curvilinear velocity (P = 0.01). This study concludes that fresh and frozen thawed semen from patients with malignant tumors is poor in quality but is still adequate for assisted reproductive techniques. As cancer therapy may further impair semen quality, patients should be offered the chance to bank sperm before undergoing cancer therapy. PMID- 11011970 TI - Frontal lobe tasks do not reflect frontal lobe function in patients with probable Alzheimer's disease. AB - Thirty-one patients with probable Alzheimer's disease (AD) according to NINCDS ADRDA criteria were psychometrically tested with various frontal lobe tasks. The results were correlated with regional cerebral glucose metabolism (rCMRG1) as measured by positron emission tomography of 18F-2-fluoro-2-deoxy-D-glucose. RCMRG1 of frontal functional-anatomically defined regions was not linked to the performance seen in frontal lobe testing. The majority of the frontal lobe tasks showed a high correlation to severity of dementia that was related to rCMRG1 of the temporo-parietal cortex. There were high intercorrelations of frontal lobe test scores to other tests. Thus, these tasks seem to measure nonspecific cognitive changes in AD patients. PMID- 11011971 TI - Hoffmann reflex profiles and strength ratios in postoperative anterior cruciate ligament reconstruction patients. AB - Rupture of the anterior cruciate ligament (ACL) typically leads to surgical reconstruction followed by an extensive rehabilitation program. One of the most commonly experienced complications associated with ACL rupture and reconstruction is quadriceps muscle atrophy. A clear understanding of the exact mechanisms associated with ACL related atrophy remains undocumented. The purpose of the present study was to investigate maximum H-reflex to maximum M-wave ratio as well as quadriceps deficit for both isometric and isokinetic peak torques in a post ACL reconstruction population. Forty subjects volunteered for participation in this study. The experimental group comprised 20 subjects who had undergone patellar tendon graft reconstruction of a torn ACL. A matched control group of 20 subjects were also measured for comparative purposes. The results indicated the control group had significantly higher quadriceps to quadriceps ratio than did the ACL group [t(38) = 9.05 p < .001]. In contrast, there was no difference in the H-max/M-max ratio for either group or leg. The strength findings of this study support previous findings. The spinal reflex results support the need for additional research in this area, specifically with more acutely injured subjects. PMID- 11011972 TI - Dynamic systems theory approach to consciousness. AB - The review of the main existing approaches to consciousness in the literature indicates that consciousness is multileveled and that humans have an ability to experience a wide range of its various states. The common views of the main approaches to consciousness are identified and considered in a three-dimensional model of states of consciousness developed by Kokoszka. This model is then described and discussed using a mathematical terms of dynamic systems theory. Since mathematics is an accepted tool for the description of the physical world, the congruence of psychological and mathematical models is understood as a meaningful support to the psychological one. Connections of the models with artificial neural networks and some applications in psychology are considered. PMID- 11011973 TI - The perception of rorschach inkblots in schizophrenia: a neural network model. AB - Schizophrenia is a psychiatric disorder characterized by a variety of cognitive deficits, including perceptual distortions and hallucinations. In recent years several studies have proposed that schizophrenia may involve a disturbance of "context". We have used a three layer neural network model constructed from an input layer followed by two computational layers to simulate responses of schizophrenic patients to the Rorschach test. In this test subjects respond to a set of ambiguous patterns created by ink blots on paper. Our model proposes that a disturbance of context caused by altered noise-to-signal ratio at the level of the single units, is responsible for schizophrenic responses to the Rorschach test. The assumption that catecholaminergic neurotransmitter systems regulate noise-to-signal ratio in cortical neurons constitutes a link between findings of altered neurotransmitter activity and deficits of cognitive functions requiring contextual integration in schizophrenia. The development of models for specific task deficits in schizophrenia could advance our insights regarding the neurological mechanisms underlying serious mental disorders such as schizophrenia. PMID- 11011974 TI - The effects of melatonin on the antioxidant systems in experimental spinal injury. AB - Melatonin has been recently shown by various in-vivo and in-vitro studies to exert potent neutralising effects on hydroxyl radicals, stimulate glutathione peroxidase (GSH-Px) activity, and protect catalase (CAT) from the destructive activity of hydroxyl radicals in neural tissue. We aimed to investigate the possible effects of pharmacological dose of melatonin on some of the antioxidant defence systems in an in-vivo study of experimental spinal injury. Seven groups of adult male Sprague Dawley rats were used in the following scheme: Group I: Naive (n = 6), Group II: Lesion (n = 8), Group III: Melatonin (n = 5), Group IV: Melatonin + Lesion (n = 8), Group V: Placebo + Lesion (n = 5), Group VI: Sham operation (n = 5), and Group VII: Placebo (n = 5). Experimental spinal injury was induced at level T7-T8 by 5 sec compression of the total cord with an aneurysm clip on anaesthetised and laminectomized animals. The total 10 mg/kg dose of melatonin (Sigma) dissolved in alcohol-water was administered i.p. four times in 2.5 mg/kg doses, at 20 min pre-, at the time of and at 1 h and 2h post compression. At 24 +/- 2h post-injury, the rats were euthanized and the lesioned segments of cord were dissected and homogenised with special care taken to distribute equal amount of injured tissue in each sample for analysis of reduced glutathione (GSH), oxidised glutathione (GSSG), superoxide dismutase (SOD), and CAT activity. Compression injury decreased GSH/GSSG ratio significantly (p < .0001). Melatonin, by itself, significantly decreased GSSG content (p < .05) and increased CAT activity (p < .05) in the naive rats. Melatonin treatment decreased GSSG activity, thus elevating GSH/GSSG ratio, and also increased SOD and CAT activity without reaching statistical significance in the lesioned animals. In conclusion, pharmacological dose of systemically applied melatonin seemed to support some features of the antioxidant defence systems in our hands. PMID- 11011975 TI - Probability and interstimulus interval effects on the N140 and the P300 components of somatosensory erps. AB - Event-related potentials (ERPs) evoked by somatosensory stimuli were studied in two experiments with manipulation of the interstimulus interval (ISI) in the range from 1 to 4 seconds and the stimulus probability from 10 to 50% in fine steps. All the stimuli were presented randomly on both index fingers as odd-ball paradigm, and the stimulus intensities on both sides were equally set at 2.5 times the subjective sensory threshold at which the stimuli could easily be ignored when delivered as nontarget stimuli. During the course of two experiments, the N140 was evoked by the target and nontarget stimuli, otherwise the P300 was evoked by the target stimuli alone. In the experiment on the effects of ISIs change, the N140 amplitude showed a tendency to increase with increasing ISI and the P300 amplitude increased with increasing ISI. In the other experiment on the effects of change in stimulus probability, the amplitudes of both the N140 and the P300 decreased with increasing stimulus probability. From the results across two experiments. it is argued that the effects of both probability and ISI changes on the amplitudes of both the N140 and the P300 do not directly reflect the recovery time of psychological functions but may also involve the complicated effects derived from the habituation of the individual generators and the difficulty for performing the task. Moreover, it is indicated that the stimulus intensity used is one of major factors to be considered for comparing results to discuss the effects of ISI and probability on the ERPs. PMID- 11011976 TI - Somatosensory evoked potentials associated with thermal activation of type II Adelta mechanoheat nociceptive afferents. AB - We evaluated evoked potentials (EPs) to noxious contact heat pulses delivered to hairy skin of healthy adults. Heat pulses from an adapting temperature of 34 degrees C to a target temperature of 52 degrees C, produced two scalp positive waves. The first peaked at 44 degrees to 45 degrees C (approximately 500 ms following stimulus onset), while the second peaked approximately 300 ms following the 52 degrees C heat pulse (approximately 1 s after stimulus onset). The first positive wave was absent from an adapting temperature of 39 degrees C, suggesting loss of synchronized activation of warm and/or low threshold mechanothermal afferents. The second EP was observed following stimulation from both adapting temperatures and was associated with subjective report of first pain. Latency difference of the pain EP from arm and leg were consistent with conduction in Adelta nociceptive afferents (approximately 10/ms). EPs to painful contact thermal stimuli may be of value in the evaluation of small fiber peripheral neuropathies and assessment of altered pain states. PMID- 11011977 TI - The relationship of neuropsychological functioning to measures of substance use in an adolescent drug abusing sample. AB - The purpose of this research was to investigate the relationship between recent and long term substance use on adolescents' neuropsychological functioning. Subjects were 77 adolescents who were referred for outpatient treatment for drug and conduct problems. Subjects were administered the Luria-Nebraska Neuropsychological Battery-III, structured interviews to assess substance use, and urine tests. Subjects were divided into neuropsychologically impaired and nonimpaired groups. Results indicated no significant group differences for: self reports of lifetime use of alcohol, cannabis, and hard drugs; self or collateral reports of recent (past 30 days) use of alcohol, cannabis, and hard drugs; or urinalysis detection of alcohol, cannabis, and hard drugs. Correlations between these dependent measures of substance use and neuropsychological functioning were also nonsignificant (all ps > .05). Although neuropsychological impairment was observed for some subjects in this sample, it was unrelated to their cannabis, hard drug, or alcohol use. These results are consistent with those found in the adult literature, given the relatively short period of time that these youth have used such substances. However, given the severity of their absolute levels of substance usage, they may be at greater risk for developing future neuropsychological problems, related directly to the ingestion of alcohol and illicit drugs, and to their secondary effects (e.g., head traumas, malnutrition). PMID- 11011978 TI - Factor structure of nonverbal cognition. AB - In order to define the factor structure of nonverbal cognitive processes, 156 twenty to sixty year-old participants were selected in Medellin (Colombia). A neuropsychological test battery for assessing different nonverbal cognitive domains (attention, memory, visuoperceptual and visuoconstructive abilities. executive functions, praxis abilities, and written calculation abilities) was administered. Initially, independent factor analyses were carried out for each domain. Three attention factors (Sustained Attention, Divided Attention, and Processing Speed, 73.1% of the variance); two memory factors (Categorical and Non Categorical Memory, 59.7% of the variance): two visuoperceptual and visuoconstructive factors (Sequential and Simultaneous, 54.0% of the variance); and two executive function factors (Categorization and Trial Error, 82.0% of the variance) were found. Further, several sequential factor analyses using Varimax orthogonal rotations for noncorrelated variables were performed. The 32 test variables were included, but progressively some variables were removed. This procedure finally selected 13 variables corresponding to five factors accounting for 72.6% of variance. Factor I was an Executive Function factor (30% of variance). Factor 2 corresponded to a Sequential Constructional factor (14.7%). Factor 3 represented a Processing Speed factor and accounted for 10.6% of the variance. Factor 4 was Visuoperceptual factor (9.5% of the variance). Finally, Factor 5 (7.8% of the variance) was a Nonverbal Memory factor. It was concluded that several, different cognitive dimensions are included in nonverbal cognition. PMID- 11011979 TI - Quantitative correlation between initial dissolution rate and heat of fusion of drug substance. AB - The initial dissolution rates of amorphous, partial crystalline and crystalline samples of terfenadine polymorphs (forms I and II) were measured by the rotating disk method. The heats of fusion due to crystalline fraction of samples were obtained by the differential scanning calorimetry (DSC) data taking into account the heat of crystallization and the heat capacity change at glass transition during the heating process. The logarithms of initial dissolution rates of different crystallinity samples were linearly correlated with the corrected heats of fusion, irrespective of the crystal forms. PMID- 11011980 TI - Preparation of enteric coated timed-release press-coated tablets and evaluation of their function by in vitro and in vivo tests for colon targeting. AB - As a new oral drug delivery system for colon targeting, enteric coated timed release press-coated tablets (ETP tablets) were developed by coating enteric polymer on timed-release press-coated tablets composed of an outer shell of hydroxypropylcellulose and core tablet containing diltiazem hydrochloride (DIL) as a model drug. The results of the in vitro dissolution tests in JP 1st fluid (pH 1.2) and JP 2nd fluid (pH 6.8) indicated that these tablets showed both acid resistance and timed-release. To clarify whether ETP tablets could have been of use in the gastrointestinal tract, ETP tablets with a layer of phenylpropanolamine hydrochloride (PPA) (a marker of gastric emptying) between the enteric coating layer and outer shell were prepared, and were administered to beagle dogs. The gastric emptying time and lag time after gastric emptying were evaluated by determining the times at which PPA and DIL first appeared in the plasma (TFA(PPA) and TFA(DIL), respectively). TFA(PPA) and TFA(DIL) were about 4 and 7 h, respectively. This value of TFA(PPA) indicated that ETP tablets displayed acid resistance in the stomach as well as in JP Ist fluid. Subtraction of TFA(PPA) from TFA(DIL) gave a value of about 3 h which agreed well with the lag time determined by in vitro dissolution test in JP 2nd fluid. Also, the results seemed to be in accordance with the time at which the tablets reached the colon after gastric emptying. Therefore, ETP tablets seemed to be an effective tool for oral site-specific delivery including targeting of the colon. PMID- 11011981 TI - Determination of plasma concentrations of losartan in patients by HPLC using solid phase extraction and UV detection. AB - PURPOSE: To establish a HPLC assay for plasma losartan and its active metabolite EXP3174 to facilitate clinical pharmacokinetic studies. METHODS: the HPLC system consisted of a 250 x 2 mm i.d. C18 reversed phase column preceded by a 4 x 4 mm guard column, a UV detector set at 254 nm, and an integrator. The mobile phase was a mixture of 0.01 M ammonium phosphate: acetonitrile: methanol (6:3:1) containing 0.02 % sodium azide and 0.04% TEA, with pH adjusted to 3.2. The system was operated isocratically at ambient temperature at a flow rate of 0.3 ml/min. Losartan and its active metabolite EXP3174 were extracted from plasma using C2 bonded silica gel standard solid phase extraction. RESULTS: recoveries of losartan and EXP3174 from plasma were greater than 70%. Using 0.5 ml of plasma sample, standard curves were linear from 10 to 300 ng/ml (r2 = 0.996 and 0.997 for losartan and EXP 3174, respectively). Sensitivity of the assay was < 10 ng/ml. Intra-and inter-assay variations were < 10 and 15%. respectively. The assay has been successfully applied to measuring plasma concentrations of losartan and EXP3174 in patients receiving a daily dose of losartan (50-100 mg). CONCLUSION: The HPLC assay has adequate sensitivity, reproducibility, and specificity for clinical pharmacokinetic studies. PMID- 11011982 TI - Plasticisation of amylodextrin by moisture: consequences for drug release from tablets. AB - Moisture influences the consolidation behaviour of amylodextrin powders and the porosity and mechanical strength of compacts thereof. The aim of this study is to relate moisture content and compact properties to drug release characteristics of amylodextrin tablets. Therefore, amylodextrin tablets containing theophylline monohydrate were prepared and their release characteristics were studied as a function of moisture content and initial porosity. Drug release from amylodextrin tablets occurs through a leaching mechanism in which cracks are progressively formed in the hydrated part of the matrix leading to almost constant release rates. Small variations in moisture content resulted in large changes of the release rate. A unique relationship between porosity and release rate, which was independent on moisture content and compaction pressure, was observed. Above a critical porosity of 0.075 crack formation was followed by disintegration and fast release. Below this critical porosity, tablets stayed intact despite of the formation of cracks, and sustained release was observed. It is concluded that control over moisture content is essential for the production of amylodextrin tablets with reproducible release characteristics. Using amylodextrin containing 10-17%, moisture, tablets with a constant release behaviour can be obtained if sufficient compaction pressure ( > 300 MPa) is applied. Lubrication of amylodextrin powders reduces the effect of porosity significantly and improves the robustness of amylodextrin tablets as a release controlling excipient in tablets largely. PMID- 11011983 TI - Intranasal administration of different liquid formulations of bumetanide to rabbits. AB - The bioavailability of bumetanide in rabbits after intranasal administration of eight formulations intended for use in acute situations has been studied. The vehicles tested were combinations of phosphate buffer, pH 7.4, glycofurol 75. polyethylene glycol 200 and coconut oil. A mixture of 51% glycofurol in polyethylene glycol 200 was administered containing doses of 1 and 8 mg bumetanide respectively. For all other formulations the lower dose level only was studied. The tmax obtained ranged from 3 to 10 min. The vehicles resulting in the highest rate of absorption were 60% glycofurol in coconut oil and pure glycofurol. The observed bioavailability for the different formulations ranged from 16 to 37% for the time period 0-120 min. The bioavailability was also calculated omitting the initial peak seen after i.v. injection, which may be undesirable. Using this method bioavailabilities of 33-82, for the time interval 5-120 min was found. The study also demonstrated that the total amount of bumetanide absorbed increased proportionally to the dose administered. The rate of absorption of bumetanide from all formulations tested may be relevant for the treatment of acute oedematous states. The tmax obtained after intranasal administration was shorter than reported for other non-parenteral routes of administration. PMID- 11011984 TI - Fabrication of oral acid resistant Fermosorb (Ferment + sorbent)-type preparations can be simplified using industrial Bacillus subtilis culture filtrates as an enzyme source. AB - In our recent article [Biziulevicius, G.A. and Zukaite, V., 1999. Int. J. Pharm. 189, 43-55] we described a novel approach in design of oral preparations intented for intestinal delivery of enzymes, based on reversible immobilization of the latter onto the polymer matrix. Fermosorb (ferment + sorbent)-type preparations, produced in such a way, are characterized as two-component delayed-release enzyme formulations being stable at acidic pH and thus ensuring the protection of an active substance in the environment of the gastric region and liberating the active substance through dissociation of the enzyme-polymer complex at neutral pH values characteristic for the intestines. In the present paper we report our updated findings showing that the technology of fabrication of two Fermosorb-type preparations manufactured (namely Fermosorb and Polyferm) can be simplified (as well as their production cost reduced) substituting the acetone precipitated enzyme preparation solutions, currently used as an enzyme source, by their precursors--industrial Bacillus subtilis culture filtrates. Moreover, we give a description of how one more Fermosorb-type preparation aimed at intestinal delivery of amylolytic enzymes can be produced in a similar manner. Determination of immobilization conditions has revealed, that irrespectively of the enzyme origin (lytic, proteolytic or amylolytic) and its source (1%, acetone precipitated preparation solution or culture filtrate), optimal for immobilization v/w ratio of the liquid phase and the polymer matrix (Biocarb L) remains the same and is equal to 10:1 (approximate). The main differences have been found to be in optimal for immobilization pH values as well as process duration in regard to the two enzyme sources applied. In case of proteolytic and amylolytic enzymes only one of the variables was different (process duration for the first ones, optimal pH for the second ones), while in case of lytic enzymes both variables were different. The percentage of the enzymes activity uptaken from the reaction mixture formed by either of the enzyme sources and the polymer matrix (approximately 60%) as well as activity losses at drying the enzyme polymer complexes ( approximately 20%), tolling in the final activity yield of near 40%, were found to be very similar. PMID- 11011985 TI - Effect of dosing time on the total intestinal transit time of non-disintegrating systems. AB - The total gastrointestinal transit time of nondisintegrating tablets may be affected by dosing time; available literature on this topic is inconclusive. OROS systems are nondisintegrating osmotically driven tablets that release drug over a period of time during their transit through the gastrointestinal tract and are excreted intact in the feces. Total transit times following morning administration of OROS systems pooled from various studies (n = 1,163 systems) showed a distribution with peak frequencies clustering around 24 and 48 h and following night administration (n = 80 systems) was found to cluster around 12 and 36 h. The total transit time distribution appears to be different following morning and night administration. However, on reanalyzing the data considering clock time when the tablet was collected rather than time post-administration, most of the difference between the distribution patterns disappeared. This suggested that total transit times after morning or night administration may be related to the bowel movement habits of the study population. Therefore, OROS systems total transit time were compared to the intrinsic bowel movement pattern of the general population reported in the literature and indeed a good correlation was seen between the two. The total transit time appears to be determined by two factors: the defecation frequency and the probability of its inclusion in the defecation event which is related to its location in the GI tract. A tablet is more likely to be excreted if it is further down in the GI tract. The total transit time data for OROS systems suggest that with the morning dosing the tablet is more likely to be excreted in the bowel movement the next morning. With the night time dosing the tablet may not be far enough in the colon to be excreted in the next morning bowel movement and therefore, it is more likely to be excreted the following morning. PMID- 11011986 TI - Application of acid-treated yeast cell wall (AYC) as a pharmaceutical additive I. AYC as a novel coating material. AB - Acid-treated yeast cell wall (AYC) was newly prepared by acidifying the cell wall of brewer's yeast and the potential to use AYC as a novel coating material was studied. AYC had an oval shape with the diameter of several microm. The rheogram of AYC aqueous dispersion showed the plastic fluid property that is generally observed in the suspension. Core tablets containing 3% of acetaminophen (AAP) were coated with the AYC aqueous dispersion containing 5% (w/v) of AYC and 0.35% (w/v) of glycerol at various coating percents. The AAP release profile from the AYC-coated tablets was studied by the JP13 paddle method using solutions at various pH. Tensile strength and permeability of oxygen and water vapor of AYC cast film were measured. The AAP release from the AYC-coated tablets showed sigmoidal release profile with an initial lag time and the duration of the lag time depended on the coating percent of AYC. The pH of the dissolution fluid or the storage at room temperature for 120 days had little affect on AAP release from the AYC-coated tablets. These results suggest that it is possible to control the start time of medicine release independent of the pH by coating of AYC, that is the time-controlled release. The AYC cast film showed a large tensile strength and an extremely small oxygen permeability coefficient and a sufficient level of water permeability coefficient in order to protect from moisture. These results present that AYC has the high utility as a novel aqueous coating material for DDS preparations. PMID- 11011987 TI - Statistical assessment of between batch stability equivalence. AB - A statistical method for testing the equivalence between batches regarding their stability is proposed. This method is based on the statistical linear model making use of a set of dummy variables to code the different batches. The method gives us the point estimates of the slope and zero intercept of one batch, and the differences and the corresponding confidence intervals with the remaining batches. In a second step, zero intercepts and slopes are estimated for all the batches. Stability equivalence assessment is based on the comparison of the confidence intervals for the differences between batches with the maximum difference allowable. The main advantages of this method are the possibility to compare several batches, to disclose the equivalence stability criteria from the statistical hypothesis about the equality between slopes, and the joint estimated of the residual variance whatever the decision to pool or not the data from different batches. This method is illustrated with two data set; the first one, previously published by other authors, involved six batches; the second data set include two batches and arose in a stability study of a commercial human insulin conducted in our laboratory. PMID- 11011988 TI - An investigation of some of the factors influencing the jet nebulisation of liposomes. AB - Multilamellar egg phosphatidylcholine liposomes with or without cholesterol have been aerosolised using four jet nebulisers. The size of aerosols generated from liposome suspensions, as measured by laser diffraction, was independent of liposome size and bilayer composition. However, increasing the phospholipid concentration caused an increase in the median size of the secondary aerosol size, although the extent of this effect was dependent on the design on the nebuliser. The total mass output of liposomal aerosols was similar for the Pari LC and Sidestream nebulisers, though the rate of output was higher for the Sidestream. In both cases, increasing lipid concentration produced a reduced rate of aerosol output. For all the nebulisers studied, a size selective process was found, resulting in the retention of the largest liposomes. PMID- 11011989 TI - Characterization and in vitro release of methotrexate from gelatin/methotrexate conjugates formed using different preparation variables. AB - The purpose of this study was to evaluate effects of preparation variables on the composition of gelatin-methotrexate conjugates, and to evaluate their in vitro stability. Conjugation variables of pH, amount of conjugating agent 1-ethyl-3 (diaminopropyl)carbodiimide HCl (EDC), and methotrexate (MTX), with unfractionated gelatin were examined. Conjugate composition was determined spectrophotometrically. The molar ratios of MTX to gelatin in the conjugates ranged from 5.9 to 64.9. Molar ratios increased with molecular weight (MW) of gelatin in the conjugate, but the weight ratio was constant. This common conjugating procedure, however, produces by-product crosslinking and produces a mix of covalent MTX binding to carboxyl and amino groups of the gelatin. For release studies, gelatin was fractionated by size exclusion spectra (SEC) into MW of 21, 91, and 195 kDa prior to conjugation. MTX release from conjugates in dialysis cassettes at 25, 37, and 50 degrees C, in isotonic pH 7.4, buffer over 72 h was assayed by high performance liquid chromatography (HPLC). There was no effect of gelatin MW on MTX release. MTX release was approximately linear and attained 2.3, 7.2, and 13% by 72 h at 25, 37, and 50 degrees C, respectively, for the 91 kDa conjugates. First-order release rate constants were 0.23 x 10(-3), 0.95 x 10(-3), and 1.8 x 10(-3) x h(-1), respectively. The calculated activation energy for MTX release was 15.8 kcal/mol. Rate constants and the activation energy for MTX release are consistent with hydrolysis of a peptide bond. Non degraded MTX species were found in the release medium at amounts similar to free MTX and were attributed to MTX polymers and MTX/gelatin fragments < 10 kDa. PMID- 11011990 TI - Penetration of topical and oral ofloxacin into the aqueous and vitreous humor of inflamed rabbit eyes. AB - PURPOSE: This study aimed to investigate the penetration of topical and oral ofloxacin into aqueous humor and vitreous humor in post-traumatic endophthalmitis model in rabbits. METHODS: A standardized intraocular infection after penetrating injury was made in the right eyes of 16 rabbits. Intraocular infection was induced by intravitreal injection of a suspension of Staphylococcus aureus. The intact left eyes were maintained as controls. The animals were divided randomly into two groups. (1) In the topical group, two drops of ofloxacin 0.3% eyedrops were instilled to both eyes every 30 min for 4 h. (2) In the topical-oral group, two doses of 25 mg/kg of ofloxacin at 12-h intervals were given orally, then the protocol of the first group was applied. Aqueous and vitreous humor samples were taken 30 min after the last drop. Ofloxacin concentrations were measured by using HPLC. RESULTS: Mean aqueous levels of ofloxacin in control eyes were: 3.25 +/- 2.55 microg/ml in topical group. 4.58 +/- 5.39 microg/ml in topical-oral group. Mean aqueous levels in inflamed eyes were: 5.21 +/- 4.55 microg/ml in topical group, 10.34 +/- 8.88 microg/ml in topical-oral group. Mean vitreous levels of ofloxacin in control eyes were: 0.17 +/- 0.07 microg/ml in topical group, 1.30 +/ 1.23 microg/ml in topical-oral group. Mean vitreous levels in inflamed eyes were: 0.35 +/- 0.22 microg/ml in topical group, 3.48 +/- 2.69 microg/ml in topical-oral group. There was no significant difference among the groups (P > 0.05), however. CONCLUSIONS: The result of this study suggests that oral supplementation of ofloxacin to topical instillation increased the ocular levels of ofloxacin in the post-traumatic endophthalmitis model. Mean drug concentrations in aqueous and vitreous humors were above the 90% minimum inhibitory concentrations (MIC90) for most of the common microorganisms causing endophthalmitis in all eyes, except in the vitreous humors of the intact eyes instilled topically. PMID- 11011991 TI - The effects of prolonged acute use and inflammation on the ocular penetration of topical ciprofloxacin. AB - PURPOSE: To study the aqueous and vitreous penetration of ciprofloxacin after prolonged acute topical administration and to investigate the effects of inflammation on drug penetration. METHODS: A standardized model of intraocular infection after penetrating injury was made in the right eyes of eight rabbits. The intact left eyes were maintained as the control. Two drops of ciprofloxacin 0.3% eyedrops were instilled topically every 1 h for 7 h to all eyes of the rabbits. Aqueous and vitreous samples (100 microl) were obtained half an hour after the last drop. Instillation was continued for 7 h more and samples were obtained as before. Drug concentrations were measured using HPLC. RESULTS: The mean aqueous humor levels of ciprofloxacin were: in control eyes 1.31 +/- 0.78 microg/ml after 7 h and 1.85 +/- 1.69 microg/ml after 14 h of instillation: in inflamed eyes 2.18 +/- 1.02 microg/ml after 7 h and 2.91 +/- 2.12 microg/ml after 14 h. The mean vitreous humor levels were: in control eyes 0.65 +/- 0.44 microg/ml after 7 h and 0.72 +/- 0.8 microg/ml after 14 h of instillation; in inflamed eyes 0.67 +/- 0.77 microg/ml after 7 h and 1.01 +/- 0.43 microg/ml after 14 h. However, the differences among the groups were not significant (P > 0.05). CONCLUSIONS: Ciprofloxacin penetration into aqueous humor was higher in 14-h topical application than that for 7 h. Inflammation increased the penetration of topical ciprofloxacin into aqueous while administered for 7 h and into both aqueous and vitreous humor while administered for 14 h. c PMID- 11011992 TI - Synthesis and in vivo evaluation of [11C]ICI 118551 as a putative subtype selective beta2-adrenergic radioligand. AB - Erytro-(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[ iso-propylamino]-2 butanol (ICI 118551) a potent clinically used beta2 adrenergic antagonist, was labelled with carbon-11 (t1/2 = 20.4 min) as a potential radioligand for the non invasive assessment of beta2 adrenergic receptors in the lung with positron emission tomography (PET). The radiolabelled compound was prepared by reductive N alkylation of its des-isopropyl precursor with [2-11C]acetone. (+/-)-[11C]ICI 118551 was obtained in greater than 98% radiochemical purity in 30 min with a radiochemical yield of 15 + 5% (non-decay corrected) and a specific radioactivity 2.5 +/- 0.5 Ci/micromol. The biological evaluation of racemic erythro (+/-) [11C]ICI 118551 in rats and Macaca Nemestrina shows a high radioactivity uptake in lung and heart. However, in both animal models no detectable displacement of lung radioactivity concentration was observed after pre-treatment with propranolol or ICI 118551, which indicates that in this organ, radioligand uptake is mostly due to non-specific binding. The biological data suggest that erythro (+/-)-[11C]ICI 118551 is not adequate to be further developed as a tracer for beta2 adrenergic receptor imaging in vivo. PMID- 11011993 TI - Gastric retention and stability of lipidized Bowman-Birk protease inhibitor in mice. AB - Bowman-Birk protease inhibitor (BBI) was modified with a reversible lipidizing agent. The palmitoylated product, Pal-BBI, and BBI were iodinated and orally administered to mice using a gavage needle. A prolonged retention of Pal-BBI was found in the stomach. Furthermore, a significant amount of Pal-BBI was detected as intact polypeptide in the stomach of mice fed with Pal-BBI, while only degradation products were detected with BBI. There was also a significant increase of radioactivity in the blood and liver in mice 1.5 h post administration of Pal-BBI. These results indicate that lipidized polypeptide can have a longer retention and lower digestion in the stomach. They also suggest that the Pal-BBI may have a higher gastrointestinal absorption than the original polypeptide. PMID- 11011994 TI - Effects of liquid phase migration on extrusion of microcrystalline cellulose pastes. AB - The behaviour of water-based microcrystalline cellulose pastes undergoing ram extrusion has been investigated. Factors affecting the redistribution of water within the extruding paste and the upstream barrel compact, such as the initial water content, extrusion rate and die geometry, have been considered. The rates of dewatering for these given systems were characterised by the gradients of the extrusion pressure-ram displacement profiles. A linear relationship between the pressure-displacement gradient and the inverse square root of the paste velocity was obtained for a given paste and extrusion geometry. At velocities where water migration was significant, the extrudate was found to have a higher water content than that of the paste in the barrel at any given time; both the extrudate and the barrel paste decreased in water content with increasing ram displacement. Spheronisation of extrudate samples has shown that the redistribution of liquid during extrusion is an important factor affecting the quality of the spheres. A paste flow model, incorporating pseudo-plastic and shear deformation terms, was used to predict the change in extrusion pressures caused by liquid phase migration. The model parameters were obtained as functions of water content and gave good agreement with the experimental extrusion profiles. PMID- 11011995 TI - Oral mucosal bioadhesive tablets of pectin and HPMC: in vitro and in vivo evaluation. AB - The potential of tablets containing 1:4, 1:1 and 4:1 weight ratios of pectin and hydroxypropyl methylcellulose (HPMC) for the sustained release of diltiazem by sublingual administration has been investigated. Measurements of maximum adhesive force to rat peritoneal membrane indicated a satisfactory bioadhesive strength. An in vitro sustained release of diltiazem over 5 h was achieved with bilayer tablets composed of a drug-free ethylcellulose layer in addition to the pectin/HPMC layer containing drug. Plasma concentration-time curves obtained following sublingual administration to rabbits of single and bilayer tablets with 1:1 weight ratios of pectin and HPMC showed evidence of sustained release of diltiazem. Bioavailability of diltiazem was 2.5 times that achieved by oral administration for single layer tablets and 1.8 times for the bilayered tablets. PMID- 11011996 TI - Hydrogen bonding in salicylsalicylic acid (salsalate) crystals. AB - An X-ray crystallographic study of the drug salsalicylic acid (salsalate) has been performed. Crystal formation of the drug is influenced by both inter- and intra-molecular hydrogen bonding. In addition an OH group in salsalate can occupy alternate ortho positions resulting in two hydrogen bonding motifs within a single crystal. PMID- 11011997 TI - Combined effect of oleic acid and polyethylene glycol 200 on buccal permeation of [D-ala2, D-leu5]enkephalin from a cubic phase of glyceryl monooleate. AB - The combined use of the lipophilic permeation enhancer, oleic acid together with polyethylene glycol 200 (PEG 200) as a co-enhancer and incorporated into the cubic liquid crystalline phase of glyceryl monooleate was investigated in the ex vivo buccal permeation of [D-Ala2, D-Leu5]enkephalin (DADLE) through porcine buccal mucosa mounted in a Franz cell. The addition of oleic acid (1%) and PEG 200 (1-10%) did not change the intact appearance of the cubic phase. PEG 200 increased the aqueous solubility of oleic acid incorporated into the cubic phase and hence promoted the transport of oleic acid into the porcine buccal mucosa. The solubilising effect of PEG 200 on oleic acid incorporated into the cubic phase was dependent on the PEG 200 concentration but was non-linear. The buccal permeation flux of DADLE significantly increased when 5% (P<0.01) or 10% (P<0.001) of PEG 200 was co-administered with 1% oleic acid compared with the cubic phase containing 1% oleic acid alone. The present results suggest that PEG 200 enhances the action of the lipophilic permeation enhancer oleic acid and that the combination of oleic acid and PEG 200 as a co-enhancer can be a useful tool to improve the membrane permeability in the buccal delivery of peptide drugs using a cubic liquid crystalline phase of glyceryl monooleate and water. PMID- 11011999 TI - Investigation of drug release from suspension using FTIR-ATR technique: part II. Determination of dissolution coefficient of drugs. AB - Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy was applied to a release experiment in order to determine the dissolution coefficient of drug particles in heterogeneous semisolid formulations. The drug release experiment was carried out using ketoconazole suspended in Vaseline with various amounts of paraffinum liquidum as donor and an artificial dodecanol collodion (DDC) membrane as acceptor compartment. Monitoring changes in IR bands due to ketoconazole the decrease of the drug content near the ATR crystal ointment was followed as a function of time. A mathematical model based on Fick's second law with a source term was used to derive the apparent dissolution coefficient Kdis by numerical fitting the experimental data. It was found that Kdis is dependent on the fraction of paraffinum liquidum in the suspension. Taking into account all experimental parameters required, the transport process was simulated and discussed in terms of drug concentration- time and drug concentration- distance profiles. Calculating the area under the mass-time curve it was tried to predict the 'dermal bioavailability' in the acceptor (AUCa). PMID- 11011998 TI - Investigation of drug release from suspension using FTIR-ATR technique: part I. Determination of effective diffusion coefficient of drugs. AB - Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy was used to study directly the release of drug particles (ketoconazole) in a liquid medium (paraffinum liquidum). In the case of the release experiment, the formulation is placed on the ATR crystal and the acceptor membrane on the top of the ointment. The decrease of the drug content in the sediment near the interface ATR crystal-formulation in the course of the release process was quantified by monitoring the changes of the IR spectrum in relevant spectral ranges using multivariate analysis. A mathematical model based on Fick's second law with appropriate initial and boundary conditions was applied in order to determine the diffusion coefficient of the drug in the liquid medium. Knowing this value, it is possible to calculate the effective diffusion coefficient of the drug in heterogeneous semisolid formulation (Vaseline) as a function of the volume fraction of the solid phase. PMID- 11012000 TI - Skin delivery of oestradiol from lipid vesicles: importance of liposome structure. AB - The aim of this study was to investigate the importance of liposome structure in oestradiol skin delivery as a tool for understanding the delivery mechanism from lipid vesicles. Liposomes of phosphatidylcholine (PC) (1), PC, sodium cholate; 86:14 w/w (II), PC, Span 80; 86.7:13.3 w/w (III) and PC, oleic acid: 84:16 w/w (IV) with 1 mg/ml radiolabelled drug were prepared. Saturated radiolabelled oestradiol solutions containing the components of I-IV were separately prepared in 90% w/w propylene glycol in water. In addition, saturated solutions containing cholate, Span, oleic acid and ethanol at the same concentrations used in vesicles were formulated. Oestradiol permeation through human epidermis was studied. Formulations I-IV increased oestradiol flux by 8.6, 17, 17 and 13-fold when used as vesicles compared with control and by 2.9. 4.0, 4.7 and 6.9-fold when used in solution with drug. Testing individual components in solution, relative fluxes were 2.9, 0.87, 1.1, 2.9 and 1.1 for PC, cholate. Span, oleic acid and 7% ethanol, respectively. Accordingly, it is important to prepare phospholipids as vesicles for efficient oestradiol skin delivery even after inclusion of oleic acid. Penetration enhancement is not the main mechanism for improved flux. Liposome components in solution have additive effect with a possible synergism in some cases. PMID- 11012001 TI - Design and in vitro evaluation of adhesive matrix for transdermal delivery of propranolol. AB - Propranolol hydrochloride, a water-soluble drug, was incorporated in three transdermal delivery systems using three polymers (hydroxypropylmethylcellulose, polyisobutylene and Ucecryl MC808). The influence of different factors (polymeric material, matrix thickness, drug content, thickness of the adhesive layer and presence of a dissolution enhancer) was investigated. Microscopic observations and DSC thermograms have permitted to demonstrate that propranolol was essentially dissolved in the HPMC matrix and dispersed in the two other matrix types. In vitro dissolution study was carried out according to European Pharmacopoeia. Release from HPMC matrices without adhesive coating was fast. Release from these matrices became more regular (reduction of the burst effect) and slow when they are coated with a 12 microm thick Ucecryl layer. Release from different PIB matrices was too slow to be suitable as TDDS for propranolol. The best release modulation was obtained from Ucecryl matrices. In all matrices types, propylene glycol accelerated propranolol release rate. The kinetic of drug release from most matrix types was more closely described by the square-root model (Higuchi). PMID- 11012002 TI - T cells reactive to keratinocyte antigens are generated during induction of contact hypersensitivity in mice. A model for autoeczematization in humans? AB - BACKGROUND: The role of keratinocytes (KC) in contact hypersensitivity (CH) has been examined more with respect to cytokine secretion and tolerance induction and less as a source of antigenic proteins to which chemical haptens can conjugate. OBJECTIVE: To determine whether KC-derived proteins can serve as antigenic carriers for haptens such as dinitrofluorobenzene (DNFB). METHODS: We examined the capacity of draining lymph node cells from BALB/c mice sensitized to DNFB to proliferate in response to hapten or to hapten-conjugated protein extracts derived from a KC line (DNP-Pam KC extract). Using limiting dilution microculture of these lymph node cells, we established DNP-specific T cell clones as well as DNP-Pam KC extract-reactive T cell clones. We also examined the proliferative responses of the DNP-Pam KC extract-reactive clones and of lymph node cells from mice sensitized to different haptens. RESULTS: Lymph node cells from DNFB sensitized mice proliferated well to hapten or to DNP-Pam KC extract. Six la(d) restricted, alphabeta TCR-bearing, CD4+ clones were established: 4 proliferated specifically to soluble hapten (DNBS), whereas 2 proliferated in response to DNP Pam KC extract. Surprisingly, the DNP-Pam KC extract-reactive clones proliferated as well to Pam KC extract without hapten. Lymph node cells from hapten-sensitized mice not only proliferated specifically in response to the hapten to which they were sensitized, but also proliferated to Pam KC extract without hapten. CONCLUSIONS: T cell clones generated during the induction of (CH) in mice include those reactive to hapten as well as those reactive to KC antigens independent of hapten. Analogous mechanisms in humans might account for autoreactive events such as id reactions associated with CH and angry back syndrome during patch testing. PMID- 11012003 TI - Incidence rates of occupational allergic contact dermatitis caused by metals. AB - BACKGROUND: Data on the incidence rates (IR) of occupational dermatoses are scarce. MATERIAL AND METHODS: We calculated the IR of occupational allergic contact dermatitis (ACD) caused by chromium, nickel and cobalt by occupation, during a 7-year period (1991-1997) from the data of the Finnish Register of Occupational Diseases and from the statistics on the working population in different occupations. RESULTS: A total of 2543 cases of occupational ACD were reported during 1991-1997. Cr caused 143 (5.6%) cases of ACD, Ni 176 cases (6.9%) and Co 41 cases (1.6%) of ACD. Women had greater number of occupational ACD from nickel, whereas occupational ACD from chromate and cobalt was more frequent in men. The ranking list of the IR of occupational ACD caused by Cr per 10,000 working years was (incidence rate in parenthesis) (1) tanners, fellmongers, and pelt dressers (12.20); (2) cast concrete product workers (6.94); (3) leather goods workers (4.71), (4) metal plating and coating workers (3.66); (5) bricklayers (3.44); (6) reinforcement concreters (2.79); and (7) building workers (1.32). The corresponding ranking list for Ni was (1) footwear workers (2.55); (2) machine and metal product assemblers (2.40); (3) electrical and teletechnical equipment assemblers (2.03); (4) precision instrument mechanics (1.73); (5) postal officials (1.48); (6) hairdressers, beauticians, and bath attendants (1.24); (7) industrial tailors and seamstresses (1.08); and (8) waiters in cafes and snack bars (1.04). The corresponding ranking list for Co was (1) printers (0.80); (2) turners, machinists, and toolmakers (0.36); and (3) machine and engine mechanics (0.17). CONCLUSION: The Finnish Register of Occupational Diseases forms a good basis for calculating IR. As IR illustrates the risk to become sensitized, preventive measures should be directed at occupations with the highest IR. PMID- 11012004 TI - Tolerance profile of a sterile moisturizer and moisturizing cleanser in irritated and sensitive skin. AB - BACKGROUND: This study evaluates the tolerance of preservative free sterile cleanser and sterile moisturizer in irritated and sensitive face skin. MATERIALS AND METHODS: An exploratory, open-label study using the cleanser and the moisturizer in combination was performed with 98 patients with a documented history of allergic contact dermatitis. The 2 products could each be used once or twice daily for 28 days. The assessment parameters at baseline and end of treatment (day 28) included the intensity of erythema, dryness/scaling by the investigator and subjective signs (burning, pruritus and stinging), according to a defined 4-point scale (absent to severe). In addition, a global assessment of the change from baseline and the overall tolerance of the products were performed by the investigator at the end of treatment. RESULTS: Ninety-four patients were included for the efficacy analysis and 96 patients for the safety analysis. At baseline, a majority of patients expressed some degree of erythema (63%), and dryness/scaling (56%). Fewer patients experienced subjective signs at baseline (44%). At the end of treatment, the results showed a statistically significant improvement of all the objective signs of irritated skin (P = .0001, Mac Nemar test), as well as the subjective signs of sensitive skin (P < 0.02). This was confirmed by the overall investigator assessment, showing an excellent or good response in 90% of the patients. In the safety analysis, 1 patient developed contact allergy to 1 ingredient of the test products (carbomer), and 3 patients exacerbated their dermatitis. CONCLUSION: Taken together, these results suggest that adequately formulated cosmetics might reduce both irritated and sensitive skin, with clinical improvement of dryness, erythema and stinging. PMID- 11012005 TI - Efficacy of a skin-protective foam in the treatment of chronic hand dermatitis. AB - BACKGROUND: Irritant and/or allergic hand dermatitis in individuals who are unable to avoid causative exposures is difficult to control. Usefulness of protective creams has been generally unsatisfactory. OBJECTIVE: To determine if hand dermatitis, primarily of an occupational nature, could be improved by the use of a protective foam containing dimethicone and glycerin. METHODS: Adult male and female subjects with chronic hand dermatitis for at least 12 months, felt to be either allergic, irritant, or combined in nature were given the study foam for routine application after a 2-week observation period. At 2 and 6 weeks the skin was evaluated by the investigator and subject for parameters including redness, scaling, fissuring, blistering, and pruritus on a numerical scale. A global evaluation also was performed. Photographs were taken at each visit. Usage of topical corticosteroids was recorded. No systemic therapies other than antihistamines were allowed. RESULTS: Thirty-one subjects were enrolled in the study and 28 completed it. The physician and subject's initial global evaluations averaged 6.13 and 5.48, respectively (0-10 scale). The final scores were 3.68 and 4.75, respectively (P < .001 for physician rating; P = .259 for subject rating). Topical corticosteroid usage was reduced in 16 of 30 (53.6%) subjects. Twenty-one of 30 subjects (70.0%) had improved over the course of the study. No adverse effects were noted. CONCLUSION: This protective foam greatly or moderately improved chronic hand dermatitis in a sizable number of individuals with previously uncontrolled dermatitis despite continuing in their regular occupation. PMID- 11012006 TI - Measurement of sensory irritation of the skin. AB - The sensory reaction to contact with chemical irritants has not been extensively studied. This neglect has been caused partly by a lack of understanding of the physiological basis of subjective irritation, and partly by inadequate methods of perceptual measurement. This article begins with a brief overview of the neurophysiology of cutaneous chemoreception (chemesthesis), and continues with a discussion of numerous sensory, physical, and perceptual factors that can affect the sensitivity to irritants. Then, guidelines for conducting perceptual measurements of subjective irritation that take these factors into account are offered. An alternative approach to direct chemosensory measurement also is proposed that relies on the fact that many of the cutaneous receptors that are sensitive to chemicals also are sensitive to temperature and/or mechanical stimulation (pain or itch). It is suggested that thermal and mechanical sensitivity might prove useful as measures of the severity of acute sensory irritation and as indicators of subclinical sensitization by environmental irritants. PMID- 11012008 TI - Pitfalls in patch testing. AB - Patch testing is an invaluable diagnostic tool in the evaluation of allergic contact dermatitis. While TrueTest has simplified the technique for many practitioners, there remains potential for error. We asked 4 experts to describe their approach to several dilemmas encountered in patch testing. Their responses will be helpful to both the veteran and tyro. PMID- 11012007 TI - Pruritic eruption at the site of a temporary tattoo. PMID- 11012010 TI - Amplitude degradation of time-reversed pulses in nonlinear absorbing thermoviscous fluids. AB - The linear wave equation in a lossless medium is time reversible, i.e., every solution p(x, t) has a temporal mirror solution p(x, -t). Analysis shows that time reversal also holds for the lossless nonlinear wave equation. In both cases, time-reversal invariance is violated when losses are present. For nonlinear propagation loses cannot normally be ignored; they are necessary to prevent the occurrence of multivalued waveforms. Further analysis of the nonlinear wave equation shows that amplification of a time-reversed pulse at the array elements also leads to a violation of time reversal even for lossless nonlinear acoustics. Numerical simulations are used to illustrate the effect of nonlinearity on the ability of a time-reversal system to effectively focus on a target in an absorbing fluid medium. We consider both the amplitude and arrival time of retrodirected pulses. The numerical results confirm that both shock generation (with the accompanying absorption) and amplification at the array, adversely affect the ability of a time-reversal system to form strong retrodirective sound fields. PMID- 11012009 TI - An old reaction in a new setting: the paraben paradox. PMID- 11012011 TI - Experimental and theoretical waveforms of Rayleigh waves generated by a thermoelastic laser line source. AB - An analytical model has been developed for the generation of surface acoustic (Rayleigh) waves in an isotropic solid by a thermoelastic laser line source. For a Gaussian light intensity profile, this model leads to an expression in closed form for the normal surface displacement of the Rayleigh wave either in the near field or in the far field domain. Quantitative agreement has been found for experiments carried out with an interferometric optical probe on a duraluminum plate. PMID- 11012012 TI - Wave propagation in 0-3/3-3 connectivity composites with complex microstructure. AB - This work presents a study of the properties of particulate composites. The whole range of particle volume fraction (0-1) and ideal 0-3, 3-3 and intermediate 0-3/3 3 connectivities are analysed. Two different approaches to produce a realistic model of the complex microstructure of the composites are considered. The first one is based on a random location of mono-dispersed particles in the matrix; while the second incorporates a size distribution of the particles based on experimental measurements. Different particle shapes are also considered. A commercial finite element package was used to study the propagation of acoustic plane waves through the composite materials. Due to the complexity of the problem, and as a first step, a two-dimensional model was adopted. The results obtained for the velocity of sound propagation from the finite element technique are compared with those from other theoretical approaches and with experimental data. The study validates the use of this technique to model acoustic wave propagation in 0-3/3-3 connectivity composites. In addition, the finite element calculations, along with the detailed description of the microstructure of the composite, provide valuable information about the micromechanics of the sample and the influence of the microstructure on macroscopic properties. PMID- 11012013 TI - Reflectometry using longitudinal, shear and Rayleigh waves. AB - A new technique of reflectometry using longitudinal, shear and Rayleigh waves is presented. Reflection coefficient as a function of angle incidence of an ultrasound beam with a finite beamwidth was measured for water-aluminum, water brass, and water-glass interfaces. The measured values have matched very favorably with the results of numerical calculations based on the angular spectrum of waves method. It has been shown that the speeds of longitudinal, shear and Rayleigh waves of a solid can be determined very accurately by measuring a spectacularly reflected signal versus angle of incidence. PMID- 11012014 TI - Measurements of acoustic properties of aqueous dextran solutions in the VHF/UHF range. AB - The acoustic properties of aqueous solutions of dextran are characterized in the frequency range of 70-400 MHz by the bio-ultrasonic spectroscopy system using an ultrasonic transmission comparison method. The attenuation, velocity, impedance, and density of aqueous dextran solutions, for six molecular weights in the range of 10,400-2,000,000 Da in the concentration range 520% by weight, are reported. All four parameters increase with increasing concentration. As the molecular weight increases, the attenuation coefficient increases and the velocity decreases. The precise frequency and molecular weight dependences of the acoustic properties of the solutions are readily determined by the system. PMID- 11012015 TI - Measuring Newtonian viscosity from the phase of reflected ultrasonic shear wave. AB - In this paper, an acoustic shear impedance model is employed to obtain a relation between the viscosity of a Newtonian fluid and phase characteristics of ultrasonic shear wave reflection from a solid-fluid interface. The phase and magnitude of the reflection coefficient can be decoupled in this model. The decoupling allows an independent relation between the acoustic shear impedance (viscosity-density product) and phase of the reflection coefficient. The model was experimentally verified for different fluid-solid combinations. Comparison of the results with the commonly used absolute reflection coefficient method demonstrates that phase measurement provides improved measurements. PMID- 11012016 TI - Experimental measurement of the acousto-electric interaction signal in saline solution. AB - Acoustic pressure alters local electrical conductivity in tissues and solutions. This work concerns the measurement of electrical conductivity in a liquid which is subjected to an acoustic pressure field created by a focused transducer. Measurements were made with four electrodes positioned in the ultrasonic focal zone, and the signal concerned is referred to as the acousto-electric interaction signal. A solution of sodium chloride in a measurement cell was subjected to ultrasound pressures of upto 1 MPa. It was shown that it is possible to quantitate the acousto-electric interaction signal once the ultrasonic vibration potential due to the Debye effect has been subtracted. The acousto-electric interaction signal was shown to be directly proportional to both the applied acoustic pressure and current. For the measurement cell used in this work, the interaction factor was found to be 5.3 microVmA(-1) MPa(-1). PMID- 11012017 TI - A novel approach to the aperture windowing in medical imaging. AB - A new technique is proposed to improve the lateral resolution in the conventional B-mode imaging systems, which enables a simple array aperture windowing in the transmitting mode. Amplitude shaping is performed without modifying the transmitting voltage of the array elements, but only varying the excitation pulse length from one element to another. This method presents some attractive practical advantages, and the reduction of the sidelobe energy is comparable to that attainable with a conventional aperture windowing. Parametric plots are given, which transform an amplitude apodization into a 'time apodization' for any type of transducer array. PMID- 11012018 TI - Computer-aided design, synthesis and biological assay of p-methylsulfonamido phenylethylamine analogues. AB - Class III antiarrhythmic agents selectively delay the effective refractory period (ERP) and increase the transmembrance action potential duration (APD). Based on our previous studies, a set of 17 methylsulfonamido phenylethylamine analogues were investigated by 3D-QSAR techniques of CoMFA and CoMSIA. The 3D-QSAR models proved a good predictive ability, and could describe the steric, electrostatic and hydrophobic requirements for recognition forces of the receptor site. According to the clues provided by this 3D-QSAR analysis, we designed and synthesized a series of new analogues of methanesulfonamido phenylethylamine (VIa i). Pharmacological assay indicated that the effective concentrations of delaying the functional refractory period (FRP) 10ms of these new compounds have a good correlation with the 3D-QSAR predicted values. It is remarkable that the maximal percent change of delaying FRP in microM of compound VIc is much higher than that of dofetilide. The results showed that the 3D-QSAR models are reliable. PMID- 11012019 TI - Oxygenated chalcones and bischalcones as potential antimalarial agents. AB - Oxygenated chalcones (3a,b) and bischalcones (4a-j) have been synthesized and evaluated for antimalarial activity against chloroquine sensitive and resistant strains of Plasmodium berghei in mice. Some of the screened compounds, 3a, 4c, 4e, 4f and 4i, have shown significant activity at 100 mg/kg dose against sensitive strain. PMID- 11012020 TI - Carbohydrate-modulated DNA photocleavage: design, synthesis, and evaluation of novel glycosyl anthraquinones. AB - Novel and artificial anthraquinone-carbohydrate hybrids were designed and synthesized, and found to effectively cleave DNA under irradiation with a long wavelength UV light and also exhibit cytotoxicity against HeLa S3 cells. PMID- 11012022 TI - Pyrrolo[2,3-d]pyrimidines containing an extended 5-substituent as potent and selective inhibitors of lck II. AB - Pyrrolo[2,3-d]pyrimidines containing a 5-(4-phenoxyphenyl) substituent are novel, potent and selective inhibitors of lck in vitro. Exploration of C-6 position of the pyrrolo[2,3-d]pyrimidine and the terminal phenyl group structure-activity relationship (SAR) is detailed. Compound 1 is orally active in animal models. PMID- 11012021 TI - Pyrrolo[2,3-d]pyrimidines containing an extended 5-substituent as potent and selective inhibitors of lck I. AB - Pyrrolo[2,3-d]pyrimidines containing a 5-(4-phenoxyphenyl) substituent are potent and selective inhibitors of Ick in vitro; some compounds are selective for lck over src. Data are shown for two compounds demonstrating that they are potent and selective inhibitors of IL2 production in cells. PMID- 11012023 TI - Novel antifungals based on 4-substituted imidazole: a combinatorial chemistry approach to lead discovery and optimization. AB - A series of 4-substituted imidazole sulfonamides has been prepared by solid-phase chemistry. These compounds were found to have good in vitro antifungal activity and constitute the first examples of C-linked azoles with such activity. The most potent inhibitor (30) demonstrated inhibition of key Candida strains at an in vitro concentration of < 100nM and compared favorably with in vitro potency of itraconazole. PMID- 11012024 TI - The synthesis and SAR of rhodanines as novel class C beta-lactamase inhibitors. AB - Beta-lactam antibiotics such as the cephalosporins and penicillins have diminished clinical effectiveness due to the hydrolytic activity of diverse beta lactamases, especially those in molecular classes A and C. A structure activity relationship (SAR) study of a high-throughput screening lead resulted in the discovery of a potent and selective non-beta-lactam inhibitor of class C beta lactamases. PMID- 11012025 TI - Expeditious synthesis and cytotoxic activity of new cyanoindolo[3,2-c]quinolines and benzimidazo[1,2-c]quinazolines. AB - Novel 6-cyanoindolo[3,2-c]quinoline and 6-cyanobenzimidazo[1,2-c]quinazoline derivatives have been synthesised by treatment of the appropriate aromatic amines with 4.5-dichloro-1,2,3-dithiazolium chloride 1 (Appel salt). The cytotoxicity and the effect of these compounds on cellular growth were measured. PMID- 11012026 TI - Synthesis and biological evaluation of novel piperidine carboxamide derived calpain inhibitors. AB - Calpain inhibitors which are derived from piperidine carboxamides in the P2 region were prepared and evaluated for mu-calpain inhibition. In particular, the keto amides 11f and 11j have Ki of 30 and 9 nM and display a more than 100-fold selectivity over the closely related cysteine protease cathepsin B. Furthermore, these compounds inhibit NMDA induced convulsions in mice indicating that calpain inhibition in brain results in some anticonvulsive properties. PMID- 11012027 TI - Synthesis and antiproliferative activity of some benzimidazole-4,7-dione derivatives. AB - A series of benzimidazole-4,7-diones bearing at the 2-position the thiomethyl group or the 2-pyridyl moiety has been synthesized and tested in vitro on three tumor cell lines. Two of them show a very good antiproliferative effect. Compounds 1 and 2d are more active or equiactive, respectively, than MMC against human lymphoblastic leukemia. Both compounds exhibit high activity on human non Hodgkin lymphoma. Compound 1 is non toxic at all the concentrations used in the antiproliferative assay and 2d is toxic only at high concentration. PMID- 11012028 TI - Second generation mimics of ganglioside GM1 as artificial receptors for cholera toxin: replacement of the sialic acid moiety. AB - In a program directed towards the design and synthesis of mimics of ganglioside GM1, the NeuAc recognition domain was replaced by simple hydroxy acids, and the affinity of the new ligands to the cholera toxin was determined by fluorescence spectroscopy. The (R)-lactic acid derivative 4 was found to display the highest affinity of the series (KD = 190 microM). PMID- 11012029 TI - Interaction of cyanine dyes with nucleic acids. Part 19: new method for the covalent labeling of oligonucleotides with pyrylium cyanine dyes. AB - New chemistry for the fluorescent labeling of oligonucleotides with cyanine dyes is proposed. It is based on the use of pyrylium salts as amine-specific reagents. Monomethyne pyrylium cyanine dye 1 was covalently linked to 5'-aminoalkyl modified oligonucleotide, with simultaneous conversion of the non-fluorescent dye 1 into fluorescent pyridinium cyanine structure 2. PMID- 11012030 TI - Enhanced pneumocystis carinii activity of new primaquine analogues. AB - New analogues of the venerable antimalarial drug primaquine have been synthesized and bioassayed in vivo against Pneumocystis carinii, a life-threatening infection common among immunosuppressed patients. Two of these new compounds are significantly more active than primaquine itself, and provide new information for future drug design and development in this area. PMID- 11012031 TI - Diphenylsulfone muscarinic antagonists: piperidine derivatives with high M2 selectivity and improved potency. AB - Piperidine analogues of our previously described piperazine muscarinic antagonists are described. Piperidine analogues show a distinct structure activity relationship (SAR) that differs from comparable piperazines. Compounds with high selectivity and improved potency for the M2 receptor have been identified. The lead compound, 12b, increases acetylcholine release in vivo. Compounds of this class may be useful for the treatment of cognitive disorders such as Alzheimer's disease (AD). PMID- 11012032 TI - Biologically active oligodeoxyribonucleotides. Part 12: N2-methylation of 2' deoxyguanosines enhances stability of parallel G-quadruplex and anti-HIV-1 activity. AB - 2'-Deoxyguanosine residues of a 3',5'-end-modified hexadeoxyribonucleotide (R 95288) with anti-HIV-1 activity were substituted with N2-methyl-2'-deoxyguanosine (m2dG). These modified oligodeoxyribonucleotides (ODNs) showed a 2-fold higher activity than R-95288. Also, the CD spectra of these ODNs indicated that the m2dG modification stabilized the tertiary structure of the G-quadruplex. PMID- 11012033 TI - Development of plasmin and plasma kallikrein selective inhibitors and their effect on M1 (melanoma) and HT29 cell lines. AB - trans-4-Aminomethylcyclohexanecarbonyl-Tyr(O-Pic)-octylamide (YO-2) inhibited plasmin (PL) selectively, while trans-4-aminomethylcyclohexanecarbonyl-Phe-4 carboxymethylanili de (YO-1) inhibited plasma kallikrein (PK). YO-2 induced apoptosis of M1 (melanoma) cell line and HT29 colon carcinoma cells during 24 h through activation of caspase-3, while YO-1 did not affect either cell line even during 48 h. PMID- 11012034 TI - Synthesis and antiviral activities of N-9-oxypurine 1,3-dioxolane and 1,3 oxathiolane nucleosides. AB - Two series of 1,3-dioxolanes and 1,3-oxathiolane nucleosides containing N-9 oxypurine were synthesized as potential antiviral agents. These compounds were prepared by reacting the sugar moieties with iodo- or bromotrimethylsilane, followed by treatment with a mixture of sodium hydride and the desired N-hydroxy purine base. The preparation of these N-hydroxybases was also described. No significant antiviral activity was observed against HIV, HBV, HSV-1, HSV-2, or HCMV. PMID- 11012035 TI - Construction of two-stranded alpha-helix peptides based on influenza virus M1 protein selectively bound to RNA. AB - Various 2alpha-helix peptides were designed and synthesized based on the RNA binding region of matrix protein M1 in influenza virus. The binding properties of the peptides to model ssRNA, ssDNA, dsDNA, and virus RNA were examined by the fluorescence studies of a dansyl group incorporated into the peptides. The peptide containing the hydrophilic residues of M1 RNA-binding region bound RNAs selectively. PMID- 11012036 TI - Synthesis and antiproliferative evaluation of 7-aminosubstituted pyrroloiminoquinone derivatives. AB - Coupling of five amines on the 7-methoxy-1,3,4,5-tetrahydropyrrolo[4,3,2 de]quinoline core was achieved and afforded, in particular, an opened analogue of the natural alkaloid wakayin. Evaluation of cytotoxic activity of compounds 2, 10 13 on L1210 cells afforded IC50 in the range 0.25 5.3 microM. PMID- 11012038 TI - Ain't no rhythm fast enough: EEG bands beyond beta. PMID- 11012037 TI - The synthesis and biological evaluation of a novel series of phthalazine PDE4 inhibitors I. AB - This communication describes the synthesis and in vitro evaluation of a novel and potent series of phosphodiesterase type IV (PDE4) inhibitors. The compounds described represent conformationally constrained analogues of RP 73401, Piclamilast. Preliminary evidences of reduced side effects of II compared to standards are also reported. PMID- 11012039 TI - High-frequency oscillations (20 to 120 Hz) and their role in visual processing. AB - Oscillatory firing of neurons in response to visual stimuli has been observed to occur with different frequencies at multiple levels of the visual system. In the cat retina, oscillatory firing patterns occur with frequencies in the range of 60 to 120 Hz (omega-oscillations). These millisecond-precise temporal patterns are transmitted reliably to the cortex and may provide a feed-forward mechanism of response synchronization. In the cortex, visual responses often show oscillatory patterning with frequencies between 20 and 60 Hz (gamma-oscillations), which are not phase locked to the stimulus onset and therefore do not show up in regularly averaged evoked potentials. Gamma-oscillatory responses synchronize with millisecond precision over long distances and are mediated by the reciprocal corticocortical connectivity. Modulatory systems like the ascending reticular activating system facilitate synchronization and increase the strength of gamma oscillations. During states of such functional cortical activation, the dominant frequency of the EEG is shifted from lower frequencies in the delta-/theta-range to higher frequencies in the gamma-range. Therefore, functional states indicate different degrees of temporal precision with which large neuronal populations interact. Response synchronization also depends on relations of global stimulus features. This suggests that millisecond-precise neuronal interactions serve as a fundamental mechanism for visual information processing. PMID- 11012040 TI - Ripple (approximately 200-Hz) oscillations in temporal structures. AB - Spontaneous network oscillations near 200 Hz have been described in the hippocampus and parahippocampal regions of rodents and humans. During the last decade the characteristics and the mechanisms behind these field "ripples" have been studied extensively, mainly in rodents. They occur during rest or slow-wave sleep and provide a very fast, short-lasting (approximately 50 msec) rhythmic and synchronous activation of specific projection cells and interneurons. Ripples are frequently triggered by a massive synaptic activation from the hippocampal CA3 subfield, which is called a sharp wave. Recent evidence suggests that ripples have a specific task in memory processing-namely, that they convey information stored in the hippocampus to the cortex where it can be preserved permanently. Network mechanisms involved in ripple oscillations may be relevant for understanding pathologic synchronization processes in temporal lobe epilepsy. PMID- 11012041 TI - Linking 600-Hz "spikelike" EEG/MEG wavelets ("sigma-bursts") to cellular substrates: concepts and caveats. AB - Somatosensory evoked human EEG and magnetoencephalographic (MEG) responses comprise a brief burst of low-amplitude, high-frequency (approximately 600 Hz) spikelike wavelets ("sigma-bursts") superimposed on the primary cortical response (e.g., the N20 to electrical median nerve stimulation). The recent surge of interest in these macroscopic sigma-burst responses is energized by the prospect of monitoring noninvasively, highly synchronized and rapidly repeating population spikes generated in the human thalamic and cortical somatosensory system. Thus, analyses of spike-related sigma-bursts could uniquely complement conventional low frequency EEG/MEG, reflecting mass excitatory and inhibitory postsynaptic potentials that potentially also incorporate subthreshold activities of undetermined functional relevance. Recent studies using spatiotemporal source analysis of multichannel recordings identified regional burst sources subcortically (near-thalamic) as well as cortically. At the primary somatosensory cortex, sigma-burst generators showed the well-established homuncular somatotopic ordering. Functionally, the 600-Hz burst appears to comprise multiple subcomponents with differential sensitivity to stimulus rate, intensity, sleep wake cycle, tactile interference, subject age, and certain movement disorders. A plenitude of cellular candidates contributing to burst generation at different levels can already now be envisaged, including cuneothalamic and thalamocortical relay cells, as well as cortical bursting pyramidal cells and fast-spiking inhibitory interneurons. Although cellular burst coding might serve to relay information with high efficiency, concepts to link macroscopic sigma-bursts and cellular substrates call for additional study. PMID- 11012042 TI - I-waves in motor cortex. AB - I-waves refer to high-frequency (approximately 600 Hz) repetitive discharge of corticospinal fibers produced by single-pulse stimulation of the motor cortex. First detected in animal preparations, this multiple discharge can also be recorded in humans with epidural electrodes over the spinal cord, and with recently developed noninvasive paired-pulse transcranial magnetic stimulation protocols. The exact nature of the generation of I-waves is still unclear, but there is convincing evidence that they originate in the motor cortex, mainly through activation of corticocortical projections onto corticospinal neurons. The ability to measure I-waves in human motor cortex allows one to test the integrity and excitability of the underlying corticocortical circuits in health and disease. PMID- 11012043 TI - Train duration effects on perception: sensory deficit, neglect, and cerebral lateralization. AB - The mechanisms of conscious perception are uncertain. In a preliminary study, dramatic effects of train duration on perception in a patient with right brain stroke were noted. In this study, the mechanisms of train duration on perception of peripheral somatosensory stimuli are examined. Subjects included healthy adults and patients with right brain infarctions. Train duration effects on perception were examined in relation to cerebral infarction, handedness, age, elevated peripheral threshold via bupivacaine, and impaired attention via diazepam or scopolamine. Perceptual thresholds to electrical pulses on the hand decreased as train duration increased, but only over the first several hundred milliseconds. Compared to controls, right brain stroke patients showed much greater lowering of threshold in the affected hand as train duration was extended. Age and bupivacaine elevated thresholds, but had little or no influence on train duration effects. Diazepam and scopolamine had no effect on thresholds. Thresholds were lower in the left than right hand of healthy dextral subjects, irrespective of age. Sinistral subjects had less left/right asymmetry. Increased train duration effect in patients is not explained by a primary elevation in threshold or by impaired vigilance. Lower perceptual thresholds in the left hand of healthy dextral subjects is consistent with right cerebral dominance for externally directed attention. PMID- 11012044 TI - EEG telemetry with closely spaced electrodes in frontal lobe epilepsy. AB - The use of additional electrodes (other than standard 10-20 electrodes) has proved to be extremely useful in the investigation of patients with temporal lobe epilepsy. The development of 32- and 64-channel EEG machines, along with the reformatting capabilities of digital EEG has greatly increased the possibilities in the number of electrodes and recording montages. The authors wanted to determine whether the use of closely spaced electrodes designed to increase the coverage of frontocentral regions is of benefit in the investigation of patients with frontocentral epilepsy. Patients investigated for frontocentral epilepsy underwent EEG telemetry with closely spaced electrodes based on the 10-10 nomenclature. Twenty-three patients were studied. An additional 30 minutes was required by technicians to create the montage. Unilateral frontal or frontocentral epileptic abnormalities were observed in 10 patients, independent bifrontal in 5 patients, synchronous bifrontal in 4 patients, and no EEG changes in 4 patients. In no patient did the addition of closely spaced electrodes lead to a change in the classification of the EEG. Closely spaced electrodes did not reveal focal abnormalities, which were not already apparent with 10-20 electrodes, nor did they demonstrate evidence of laterality in bilaterally synchronous discharges. PMID- 11012045 TI - Abnormal vasoreaction to arousal stimuli--an early sign of diabetic sympathetic neuropathy demonstrated by laser Doppler flowmetry. AB - Early diagnosis of diabetic autonomic neuropathy contributes to the prevention of serious complications and improves the prognosis of patients with diabetes. Common tests of peripheral autonomic function are the quantitative sudomotor axon reflex test or the sympathetic skin response (SSR). Quantitative sudomotor axon reflex test is quantifiable but technically demanding. Sympathetic skin response cannot be quantified easily. To study whether measurement of skin vasomotion is suited to assess early sympathetic peripheral neuropathy, we monitored skin blood flow at the index finger pulp using laser Doppler flowmetry before and after electrical stimulation. We assured that the stimulus was sufficient to elicit an efferent sympathetic response by monitoring palmar SSR ipsilateral to the flow measurement. In 21 diabetic patients with at least stage one polyneuropathy and 21 age-matched controls, SSR was recorded from one palm and sole following electrical stimulation at the contralateral wrist. Sympathetic skin response was present at the palms in all patients and controls and absent at the sole of two patients only. Eight patients (38.9%) had abnormal SSR, with absent plantar responses in two patients, prolonged plantar latencies in six patients, and prolonged volar SSR latencies in two patients. Skin blood flow responses were more often abnormal (46.1%) than SSR (P < 0.05), responses were delayed in two patients and absent in another 8 patients. Skin blood flow retest reliability was high with a repeatability coefficient of 10.64% in controls and 12.34 % in patients. Skin blood flow monitoring after sympathetic stimulation provides a reproducible parameter of sympathetic vasomotor control and complements the diagnostic value of SSR testing. PMID- 11012046 TI - Maximal and minimal motor conduction velocity in amyotrophic lateral sclerosis and X-linked bulbospinal muscular atrophy measured by Harayama's collision method. AB - Measurement of the maximal (Vmax) and minimal (Vmin) motor nerve conduction velocities was performed in amyotrophic lateral sclerosis (ALS), bulbospinal muscular atrophy (BSMA), and control subjects. The collision method as described initially by Harayama and coworkers was used. This allowed for the correction of the velocity recovery effect (VRE) in Hopf's original method. The purpose of this study is to clarify the controversial results regarding the Vmin and the difference between Vmax and Vmin (Vmax-Vmin) in ALS and to compare these results with BSMA, and clarify the usefulness of Harayama's method. In ALS, a reduction of Vmax and Vmin, and an increase of Vmax-Vmin were found in both median and posterior tibial nerve. In BSMA, a reduction of Vmin and an increase of Vmax-Vmin in the median nerve were noted. Some patients whose results of conventional nerve conduction study were entirely within normal range showed abnormal results in Vmin and/or Vmax-Vmin. These results suggest that the correction of VRE is essential to determine a Vmin, and motor fibers with abnormally slow conduction velocities were present in ALS and BSMA. Harayama's collision method is useful to detect abnormalities of motor fibers with submaximal conduction velocities. PMID- 11012047 TI - Radical excision of intramedullary spinal cord tumors: surgical morbidity and long-term follow-up evaluation in 164 children and young adults. AB - OBJECT: The majority of intramedullary spinal cord tumors (IMSCT) in children and young adults are low-grade gliomas. Radical resection of similar tumors in the cerebral hemisphere or cerebellum is usually curative; however, the conventional management for IMSCTs remains partial resection followed by radiotherapy because of the concern for surgical morbidity. Nevertheless, radical resection of IMSCTs without routine adjuvant treatment has been the rule at our institution since 1980. In an attempt to resolve this controversy, the long-term morbidity and survival in a large series of children have been retrospectively reviewed. METHODS: The database records and current status of 164 patients 21 years of age and younger in whom an IMSCT was resected were reviewed. A gross-total resection (>95%) was achieved in 76.8% of the surgical procedures. Subtotal resections (80 95%) were performed in 20. 1%. The majority of patients (79.3%) had histologically low-grade lesions. There were no deaths due to surgery. When comparing the preoperative and 3-month postoperative functional grades, 60.4% stayed the same, 15.8% improved, and 23.8% deteriorated. Only 13 patients deteriorated by more than one functional grade. Patients with either no deficits or only mild deficits before surgery were rarely injured by the procedure, reinforcing the importance of early diagnosis and treatment. The major determinant of long-term patient survival was histological composition of the tumor. The 5-year progression-free survival rate was 78% for patients with low grade gliomas and 30% for those with high-grade gliomas. Patients in whom an IMSCT was only partially resected (<80%) fared significantly worse. CONCLUSIONS: The long-term survival and quality of life for patients with low-grade gliomas treated by radical resection alone is comparable or superior to minimal resection and radiotherapy. The optimum therapy for patients with high-grade gliomas is yet to be determined. PMID- 11012048 TI - Nonsurgically managed patients with degenerative spondylolisthesis: a 10- to 18 year follow-up study. AB - OBJECT: Controversy exists concerning the indications for surgery and choice of surgical procedure for patients with degenerative spondylolisthesis. The goals of this study were to determine the clinical course of nonsurgically managed patients with degenerative spondylolisthesis as well as the indications for surgery. METHODS: A total of 145 nonsurgically managed patients with degenerative spondylolisthesis were examined annually for a minimum of 10 years follow-up evaluation. Radiographic changes, changes in clinical symptoms, and functional prognosis were surveyed. Progressive spondylolisthesis was observed in 49 patients (34%). There was no correlation between changes in clinical symptoms and progression of spondylolisthesis. The intervertebral spaces of the slipped segments were decreased significantly in size during follow-up examination in patients in whom no progression was found. Low-back pain improved following a decrease in the total intervertebral space size. A total of 84 (76%) of 110 patients who had no neurological deficits at initial examination remained without neurological deficit after 10 years of follow up. Twenty-nine (83%) of the 35 patients who had neurological symptoms, such as intermittent claudication or vesicorectal disorder, at initial examination and refused surgery experienced neurological deterioration. The final prognosis for these patients was very poor. CONCLUSIONS: Low-back pain was improved by restabilization. Conservative treatment is useful for patients who have low-back pain with or without pain in the lower extremities. Surgical intervention is indicated for patients with neurological symptoms including intermittent claudication or vesicorectal disorder, provided that a good functional outcome can be achieved. PMID- 11012049 TI - Incidence and outcome of kyphotic deformity following laminectomy for cervical spondylotic myelopathy. AB - OBJECT: The authors undertook a study to explore the predisposing risk factors, frequency of occurrence, and clinical implications of kyphosis following laminectomy for cervical spondylotic myelopathy (CSM). METHODS: Preoperative radiological studies were available in 46 patients with CSM who had undergone laminectomy. Records were reviewed to obtain demographic data and operative reports. Preoperative radiographs were assessed to determine spinal alignment. In a follow-up interview the authors established clinical outcome and patient satisfaction. Postoperative cervical alignment and mobility was also determined by assessing lateral neutral, flexion, and extension x-ray films. Preoperatively, the cervical spine was shown to be kyphotic in four (9%) of 46, straight in 20 (43%) of 46, and lordotic in 22 (48%) of 46 patients. Nine (21%) of 42 patients with either straight or lordotic alignment demonstrated in the preoperative period developed kyphosis after surgery. Kyphosis developed in six (30%) of 20 patients in whom straight spinal alignment was demonstrated preoperatively and in only three (14%) of 22 patients in whom lordosis was found preoperatively. Clinically, 13 (29%) of 45 patients improved and 19 (42%) of 45 remained unchanged after an average 4-year follow-up period; 36 (80%) patients believed that their surgery was successful (one patient, who was mentally retarded, could not respond to the follow-up questionnaire). Spinal alignment was not predictive of outcome; cervical mobility as demonstrated on flexion and extension, however, correlated with improved functional performance (p = 0.005). CONCLUSIONS: Kyphosis may develop in up to 21% of patients who have undergone laminectomy for CSM. Progression of the deformity appears to be more than twice as likely if preoperative radiological studies demonstrate a straight spine. In this study, clinical outcome did not correlate with either pre- or postoperative sagittal alignment. PMID- 11012050 TI - Treatment of Down syndrome-associated craniovertebral junction abnormalities. AB - OBJECT: Operative intervention for craniovertebral junction (CVJ) instability in patients with Down syndrome has become controversial, with reports of a low incidence of associated neurological dysfunction and high surgical morbidity rates. The authors analyzed their experience in light of these poor results and attempted to evaluate differences in management. METHODS: Medical and radiographic records of 36 consecutive patients with Down syndrome and CVJ abnormalities were reviewed. The most common clinical complaints included neck pain (15 patients) and torticollis (12 patients). Cervicomedullary compression was associated with ataxia and progressive weakness. Hyperreflexia was documented in a majority of patients (24 cases), and 13 patients suffered from varying degrees of quadriparesis. Upper respiratory tract infection precipitated the presentation in five patients. Four patients suffered acute neurological insults after a minor fall and two after receiving a general anesthetic agent. Atlantoaxial instability was the most common radiographically observed abnormality (23 patients), with a rotary component present in 14 patients. Occipitoatlantal instability was also frequently observed (16 patients) and was coexistent with atlantoaxial dislocation in 15 patients. Twenty individuals had bone anomalies, the most frequent of which was os odontoideum (12 patients) followed by atlantal arch hypoplasia and bifid anterior or posterior arches (eight patients). Twenty-seven patients underwent surgical procedures without subsequent neurological deterioration, and a 96% fusion rate was observed. In five of 11 patients basilar invagination was irreducible and required transoral decompression. Overall, 24 patients enjoyed good or excellent outcomes. CONCLUSIONS: The results of this series highlight the clinicopathological characteristics of CVJ instability in patients with Down syndrome and suggest that satisfactory outcomes can be achieved with low surgical morbidity rates. PMID- 11012051 TI - Single-stage anterior-posterior decompression and stabilization for complex cervical spine disorders. AB - OBJECT: To evaluate the applicability and safety of single-stage combined anterior-posterior decompression and fusion for complex cervical spine disorders, the authors retrospectively reviewed 72 consecutive procedures of this type performed using a uniform technique at a single center. METHODS: The indications for decompression and stabilization included: postlaminectomy kyphosis (15 patients), trauma (19 patients), spondylosis and congenital stenosis (32 patients), and ossification of the posterior longitudinal ligament (six patients). All patients underwent anterior cervical corpectomies in which allograft fibula and plates were placed, with 89% of patients undergoing two- or three-level procedures (range one-four levels). Lateral mass plating with autograft (morselized iliac crest) fusion was performed in all patients while the same anesthetic agent was still in effect. A hard cervical collar was used postoperatively in all patients (mean 13 weeks). All patients were followed for a minimum of 2 years (mean 29 months). Fusion was determined to be successful in all 72 patients (100%). Although the short-term morbidity rate reached 32%, the significant long-term morbidity rate was only 5%. At the 2-year follow-up examination, anterior cervical plate dislodgment was seen in one patient, and 16 of the 516 lateral mass screws implanted were observed to have partially backed out. However, there were no cases of nerve root injury, strut graft extrusion, or anterior plate or screw fracture. There were no clinically significant hardware complications and no patient required repeated operation. CONCLUSIONS: The combined single-stage anterior-posterior decompression, reconstruction, and instrumentation procedure represents a viable option in the treatment of a select group of patients with complex cervical spinal disorders. The technique provides immediate rigid stabilization of the cervical spine, prevents anterior plate failure or strut graft extrusion, and eliminates the need for halo immobilization postoperatively. Furthermore, a higher rate of fusion is achieved with this combined approach than with the anterior approach alone. PMID- 11012052 TI - A randomized prospective study of an anterior cervical interbody fusion device with a minimum of 2 years of follow-up results. AB - OBJECT: Despite variations in technique, inherent problems persist with current approaches to anterior cervical fusion. This study was performed to determine whether anterior cervical fusion performed using an investigational device was safe and effective in the treatment of degenerative cervical disc disorders and whether this device offered advantages over current techniques. METHODS: Fifty four patients with radiculopathy with or without mild myelopathy due to one- or two-level cervical degenerative disc disease were randomized as part of a Food and Drug Administration device study. Following microsurgical discectomy, the control group was treated with iliac crest graft fusion; the experimental study group underwent insertion of an interspace cage and placement of a local autograft. All patients received postoperative follow-up care for at least 2 years. Good or excellent results were found in approximately 97% of the experimental group and 88% of the control group. A solid fusion was achieved in all patients who underwent one-level cage placement, and a solid fusion at one or both levels was achieved in over 90% of both groups. Chronic donor site pain was reported by 31% of the control group. CONCLUSIONS: In this study, the use of an interbody fusion cage avoided donor site morbidity and placement of autograft achieved a high rate of good or excellent results. Interbody fusion cages appear safe and effective, and their use helps to avoid some of the inherent problems in performing current anterior cervical fusion techniques. PMID- 11012053 TI - Direct anterior screw fixation for recent and remote odontoid fractures. AB - OBJECT: The management of odontoid fractures remains controversial. Only direct anterior screw fixation provides immediate stabilization of the spine and may preserve normal C1-2 motion. To determine the indications, optimum timing, and results for direct anterior screw fixation of odontoid fractures, the authors reviewed the surgery-related outcome of patients who underwent this procedure at two institutions. METHODS: One hundred forty-seven consecutive patients (98 males and 49 females) who underwent direct anterior screw fixation for recent (< or = 6 months postinjury [129 patients]) or remote (> or = 18 months postinjury [18 patients]) Type II (138 cases) or III (nine cases) odontoid fractures at the University of Utah (94 patients) and National Institute of Traumatology in Budapest, Hungary (53 patients) between 1986 and 1998 are included in this study (mean follow up 18.2 months). Data obtained from clinical examination, review of hospital charts, operative findings, and imaging studies were used to analyze the surgery-related results in these patients. In patients with recent fractures there was an overall bone fusion rate of 88%. The rate of anatomical bone fusion of recent fractures was significantly (p < or = 0.05) higher in fractures oriented in the horizontal and posterior oblique direction (compared with anterior oblique), but this finding was independent (p > or = 0.05) of age, sex, number of screws placed (one or two), and the degree or the direction of odontoid displacement. In patients with remote fractures there was a significantly lower rate of bone fusion (25%). Overall, complications related to hardware failure occurred in 14 patients (10%) and those unrelated to hardware in three patients (2%). There was one death (1%) related to surgery. CONCLUSIONS: Direct anterior screw fixation is an effective and safe method for treating recent odontoid fractures (<6 months postinjury). It confers immediate stability, preserves C1-2 rotatory motion, and achieves a fusion rate that compares favorably with alternative treatment methods. In contradistinction, in patients with remote fractures (> or = 18 months postinjury) a significantly lower rate of fusion is found when using this technique, and these patients are believed to be poor candidates for this procedure. PMID- 11012054 TI - Spinal nerve root repair and reimplantation of avulsed ventral roots into the spinal cord after brachial plexus injury. AB - OBJECT: The authors review the first series of 10 cases in which injured intraspinal brachial plexus were surgically repaired. They describe the technique of spinal cord implantation or repair of ruptured nerve roots, as well as patient outcome. METHODS: Spinal root repair/implantation was performed from 10 days to 9 months postinjury. There were nine male patients and one female patient. Postoperatively in most cases, regeneration of motor neurons from the spinal cord to denervated muscles could be demonstrated. The first signs of regeneration were noted approximately 9 to 12 months postoperatively. Useful function with muscle power of at least Medical Research Council Grade 3 occurred in three of 10 cases. Magnetic brain stimulation studies revealed a normal amplitude and latency from the cortex to reinnervated muscles on surgically treated and control sides. A certain degree of cocontraction between antagonistic muscles (for example, biceps triceps) compromised function. With time there was a reduction of cocontractions, probably due to spinal cord plasticity. In these cases there was also, surprisingly, a return of sensory function, although the mechanism by which this occurred is uncertain. Sensory stimulation (thermal and mechanical) within the avulsed dermatomes was perceived abnormally and/or experienced at remote sites. There was some return of patients' sense of joint position. CONCLUSIONS: A short time lag between the accident and the surgery was recognized as a significant factor for a successful outcome. Reimplantation of avulsed nerve roots may be combined with other procedures such as nerve transfers in severe cases of brachial plexus injury. PMID- 11012055 TI - Anatomical projection of the cervical uncinate process in ventral, ventrolateral, and posterior decompressive surgery. AB - OBJECT: The cervical uncinate processes (UPs), their variations, and the relationships between the neurovascular structures and surrounding bone were investigated in this anatomical study. The object of this study was to highlight the important surgery-related considerations associated with ventral, ventrolateral, and posterior decompressive surgery. METHODS: Forty-nine adult C3 7 dry bone samples were used, and 10 measurements were obtained for each vertebra. The anterior measurements involved the cervical uncinate process (UP): height, width, length, distance between its tip and vertebral foramina, interuncinate process distance, sagittal angle with the superior margin of the vertebral body (VB), VB anteroposterior diameter, and VB width. Posterior measurements involved the vertical distance between the superior border of the lamina at the lamina-facet joint and the tip of the UP, as well as the horizontal distance between the medial-most border of the superior facet and the tip of the UP. All symmetrical structures were measured bilaterally. There were no statistically significant differences between right- and left-sided measurements in this series. The height of the UP increased gradually at each segmental level between C-3 and C-7. The width of the UP did not change with segmental level (5.0 mm at C-3 compared with 5.3 mm at C-7). On average, the length of the UP was relatively constant. The distance from the tip of the UP to vertebral foramina averaged 1 mm at the C2-3 level and 1.5 mm at the C5-6 level. Interuncinate distance and VB width gradually increased and were highly variable, which appeared to be related with osteophyte formation. There was a slight gradual increase from C-3 to lower segments, and it paralleled with the midline anteroposterior diameter of the same VB. The angle between the UP and the superior margin of the VB exhibited great variety. The posterior measurements decreased gradually from C-3 to C-7. CONCLUSIONS: Based on the data obtained in this study, a surgeon is provided with a three-dimensional orientation as well as anatomical knowledge. This knowledge also allows for a more effective neurovascular decompression by minimizing the surgery-related complications. PMID- 11012056 TI - In vitro biomechanical analysis of three anterior thoracolumbar implants. AB - OBJECT: The goal of this study was to evaluate the comparative efficacy of three commonly used anterior thoracolumbar implants: the anterior thoracolumbar locking plate (ATLP), the smooth-rod Kaneda (SRK), and the Z-plate. METHODS: In vitro testing was performed using the T9-L3 segments of human cadaver spines. An L-1 corpectomy was performed, and stabilization was achieved using one of three anterior devices: the ATLP in nine spines, the SRK in 10, and the Z-plate in 10. Specimens were load tested with 1.5-, 3-, 4.5-, and 6-Nm in flexion and extension, right and left lateral bending, and right and left axial rotation. Angular motion was monitored using two video cameras that tracked light-emitting diodes attached to the vertebral bodies. Testing was performed in the intact state in spines stabilized with one of the three aforementioned devices after the devices had been fatigued to 5000 cycles at +/- 3 Nm and after bilateral facetectomy. There was no difference in the stability of the intact spines with use of the three devices. There were no differences between the SRK- and Z-plate instrumented spines in any state. In extension testing, the mean angular rotation (+/- standard deviation) of spines instrumented with the SRK (4.7 +/- 3.2 degrees) and Z-plate devices (3.3 +/- 2.3 degrees) was more rigid than that observed in the ATLP-stabilized spines (9 +/- 4.8 degrees). In flexion testing after induction of fatigue, however, only the SRK (4.2 +/- 3.2 degrees) was stiffer than the ATLP (8.9 +/- 4.9 degrees). Also, in extension postfatigue, only the SRK (2.4 +/- 3.4 degrees) provided more rigid fixation than the ATLP (6.4 +/- 2.9 degrees). All three devices were equally unstable after bilateral facetectomy. The SRK and Z-plate anterior thoracolumbar implants were both more rigid than the ATLP, and of the former two the SRK was stiffer. CONCLUSIONS: The authors' results suggest that in cases in which profile and ease of application are not of paramount importance, the SRK has an advantage over the other two tested implants in achieving rigid fixation immediately postoperatively. PMID- 11012057 TI - In vitro biomechanical investigation of the stability and stress-shielding effect of lumbar interbody fusion devices. AB - OBJECT: Interbody fusion devices are rapidly gaining acceptance as a method of ensuring lumbar interbody arthrodesis. Although different types of devices have been developed, the comparative reconstruction stability remains controversial. It also remains unclear how different stress-shielded environments are created within the devices. Using a calf spine model, this study was designed to compare the construct stiffness afforded by 11 differently designed lumbar interbody fusion devices and to quantify their stress-shielding effects by measuring pressure within the devices. METHODS: Sixty-six lumbar specimens obtained from calves were subjected to anterior interbody reconstruction at L4-5 by using one of the following interbody fusion devices: four different threaded fusion cages (BAK device, BAK Proximity, Ray TFC, and Danek TIBFD), five different nonthreaded fusion devices (oval and circular Harms cages, Brantigan PLIF and ALIF cages, and InFix device); two different types of allograft (femoral ring and bone dowel) were used. Construct stiffness was evaluated in axial compression, torsion, flexion, and lateral bending. Prior to testing, a silicon elastomer was injected into the cages and intracage pressures were measured using pressure needle transducers. CONCLUSIONS: No statistical differences were observed in construct stiffness among the threaded cages and nonthreaded devices in most of the testing modalities. Threaded fusion cages demonstrated significantly lower intracage pressures compared with nonthreaded cages and structural allografts. Compared with nonthreaded cages and structural allografts, threaded fusion cages afforded equivalent reconstruction stiffness but provided more stress-shielded environment within the devices. PMID- 11012058 TI - Lasting paraplegia caused by loss of lumbar spinal cord interneurons in rats: no direct correlation with motor neuron loss. AB - OBJECT: The aims of this study were to investigate further the role played by lumbar spinal cord interneurons in the generation of locomotor activity and to develop a model of spinal cord injury suitable for testing neuron replacement strategies. METHODS: Adult rats received intraspinal injections of kainic acid (KA). Locomotion was assessed weekly for 4 weeks by using the Basso, Beattie, and Bresnahan (BBB) 21-point locomotor scale, and transcranial magnetic motor evoked potentials (MMEPs) were recorded in gastrocnemius and quadriceps muscles at 1 and 4 weeks. No changes in transcranial MMEP latency were noted following KA injection, indicating that the descending motor pathways responsible for these responses, including the alpha motor neurons, were not compromised. Rats in which KA injections included much of the L-2 segment (10 animals) showed severe locomotor deficits, with a mean BBB score of 4.5 +/- 3.6 (+/- standard deviation). Rats that received lesions rostral to the L-2 segment (four animals) were able to locomote and had a mean BBB score of 14.6 +/- 2.6. Three rats that received only one injection bilaterally centered at L-2 (three animals) had a mean BBB score of 3.2 +/- 2. Histological examination revealed variable loss of motor neurons limited to the injection site. There was no correlation between motor neuron loss and BBB score. CONCLUSIONS: Interneuron loss centered on the L 2 segment induces lasting paraplegia independent of motor neuron loss and white matter damage, supporting earlier suggestions that circuitry critical to the generator of locomotor activity (the central pattern generator) resides in this area. This injury model may prove ideal for studies of neuron replacement strategies. PMID- 11012059 TI - Effect of aluminum on neurological recovery in rats following spinal cord injury. AB - OBJECT: This investigation was undertaken to study the effect of aluminum on neurobehavioral, electrophysiological, structural, and biochemical changes in rats following spinal cord injury (SCI). METHODS: Adult male Sprague-Dawley rats classified into different groups were given aluminum sulfate-dosed drinking water in the concentrations of 0%, 0.25%, 0.5% and 1%, respectively. After 30 days of aluminum treatment, the animals were subjected to spinal cord trauma. Laminectomy was performed at T7-8 in anesthetized rats, followed by placement of a compression plate (2.2 x 5 mm) loaded with a 35-g weight over the exposed spinal cord for 5 minutes. Control animals underwent the same surgical procedure, but the compression injury was not induced (sham). Postoperative neurological function was assessed using the inclined-plane test and by obtaining a modified Tarlov score and vocal/sensory score daily for 10 days. Electrophysiological changes were assessed using corticomotor evoked potentials, whereas pathological changes were assessed by light microscopy. The level of vitamin E in the spinal cord was measured as an index of antioxidant defense. The behavioral, biochemical, and histological analyses were performed in a blinded fashion. CONCLUSIONS: Analysis of results obtained in the behavioral studies revealed that the compression of spinal cord produced transient paraparesis in which a maximum motor deficit occurred at Day 1 following SCI and resolved over a period of 10 days. Administration of aluminum significantly impaired the recovery following SCI. Analysis of the results of the biochemical, electrophysiological, and histopathological studies also confirmed the deleterious effects of aluminum on recovery from SCI in rats. PMID- 11012060 TI - New approach to cervical flexion deformity in ankylosing spondylitis. Case report. AB - The treatment of cervical fixed flexion deformity in ankylosing spondylitis presents a challenging problem that is traditionally managed by a corrective cervicothoracic osteotomy. The authors report a new approach to this problem that involves performing a two-level osteotomy at the level of maximum spinal curvature, thereby achieving complete anatomical correction in a one-stage procedure. This 48-year-old woman with ankylosing spondylitis presented with a 30 year history of progressive neck deformity that left her unable to see ahead and caused her to experience difficulty eating, drinking, and breathing on exertion. On examination, she exhibited a 90 degrees fixed flexion deformity of the cervical spine, which was maximum at C-4; this was confirmed on imaging studies. A two-level osteotomy was performed at C3-4 and C4-5 around the area of maximum spinal curvature, and the deformity was corrected by extending the head on its axis of rotation through the uncovertebral joints. The spine was stabilized using a Ransford loop. An excellent anatomical position was achieved, as was complete correction of the deformity. A two-level midcervical osteotomy performed at the level of maximum spinal curvature in ankylosing spondylitis enables complete correction of severe fixed flexion deformity in a single procedure. Preservation of the uncovertebral joints allows smooth and safe correction of the deformity about their axis of rotation. PMID- 11012061 TI - Dorsal arachnoid web with spinal cord compression: variant of an arachnoid cyst? Report of two cases. AB - Spinal arachnoid cysts are diverticula of the subarachnoid space that may compress the spinal cord; these lesions are most commonly found in the thoracic spine. Two patients who presented with thoracic myelopathy were noted on magnetic resonance imaging to have focal indentation of the dorsal thoracic cord, with syringomyelia inferior to the site of compression. Both patients were found at operation to have discrete arachnoid "webs" tenaciously attached to the dura mater and pia mater. These webs were not true arachnoid cysts, yet they blocked the flow of cerebrospinal fluid (CSF) and caused focal compression of the spinal cord. The mass effect appeared to be the result of a pressure gradient created by the obstruction of CSF flow in the dorsal aspect of the subarachnoid space. Both patients responded well to resection of the arachnoid web. Arachnoid webs appear to be rare variants of arachnoid cysts and should be suspected in patients with focal compression of the thoracic spinal cord. PMID- 11012062 TI - Calcifying pseudoneoplasms of the spine with myelopathy. Report of two cases. AB - The authors describe two cases of calcifying pseudoneoplasms, rare degenerative lesions that mimic tumor or infection. One case involved the cervical spine and the second the thoracic spine. Both patients experienced progressive myelopathy from extradural compression of the spinal cord. The radiological evaluation, pathological findings in the lesions, treatment, and follow up are described. Total or subtotal excision can relieve symptoms and prevent recurrence of this lesion. PMID- 11012063 TI - Subacute posttraumatic ascending myelopathy after spinal cord injury. Report of three cases. AB - Subacute posttraumatic ascending myelopathy is a rare disorder, unrelated to syrinx formation or mechanical instability, that may gradually emerge within the first 1 to 2 weeks after a spinal cord injury. The authors describe three patients with this syndrome and discuss its possible causes as well as its clinical presentation, imaging characteristics, treatment, and patient prognosis. PMID- 11012064 TI - Rotational compression of the vertebral artery at the point of dural penetration. Case report. AB - Vertebrobasilar insufficiency (VBI) due to vertebral artery (VA) compression occurs in a significant number of patients. Rotational compression of the VA usually occurs below C-2, where the artery is pinched during head rotation, leading to thrombus formation and subsequent cerebellar infarction. Although this problem has been reported to occur at the atlantooccipital levels, a review of the literature revealed no published cases of VA compression at the point of dural penetration, which is located above the atlantooccipital membrane. The authors report the case of a 30-year-old woman who presented with signs and symptoms of VBI. Dynamic angiography demonstrated left-sided VA compression at the site at which dural penetration had occurred, proximal to the posterior inferior cerebellar artery. Surgical decompression of the left VA at the point of dural penetration relieved the symptoms, and postoperative dynamic angiography demonstrated complete resolution of the positional compression of the left VA. Because of these findings, an additional possible location for rotational compression of the VA is described, namely, the point of dural penetration. The authors suggest a method of surgically treating rotational VA compression at this site. PMID- 11012065 TI - Embolization of spinal cord arteriovenous malformations with an ethylene vinyl alcohol copolymer dissolved in dimethyl sulfoxide (Onyx liquid embolic system). Report of two cases. AB - In this paper the authors describe the first use of a new liquid embolic agent (Onyx) to treat spinal cord arteriovenous malformations (AVMs). Because its properties make it more predictable to use than currently available liquid agents, the authors believe that this material has great potential in the endovascular management of both spinal cord and brain AVMs. This very promising agent merits further clinical study. PMID- 11012066 TI - Split cord malformation with partial eventration of the diaphragm. Case report. AB - The authors describe the case of a 3-year-old girl who presented with a dorsal split cord malformation (SCM) and was found to have eventration of the diaphragm. Although the child did not undergo surgery for eventration, its presence suggests a need for careful preoperative planning and clinical evaluation to rule out or confirm the anomalies associated with spinal dysraphism or SCM. PMID- 11012067 TI - Eosinophilic granuloma of the spine: early spontaneous disappearance of tumor detected on magnetic resonance imaging. Case report. AB - Two cases of eosinophilic granuloma (EG) of the spine associated with neurological deficits are presented. The patients were treated conservatively by using external fixation with a brace as well as bed rest. Neurological deficits and pain diminished and finally disappeared as the tumor mass decreased in size, as seen on magnetic resonance (MR) imaging. During the 5-year follow-up period no recurrence of the tumors was detected on MR images. Surgical treatment for spinal EG in children presenting with typical vertebra plana is not recommended except for those with severe or progressive palsy and for those in whom the disease requires differential diagnosis. PMID- 11012068 TI - Intraspinal familial clear cell meningioma in a mother and child. Case report. AB - The authors present a case of familial clear cell meningioma in which the proband is a child with an intraspinal tumor. The clear cell meningioma variant has recently been studied. The literature regarding clear cell meningioma is reviewed. PMID- 11012069 TI - Intraspinal extradural myxopapillary ependymoma of the sacrum arising from the filum terminale externa. Case report. AB - Extradural ependymomas of the sacrococcygeal region are very rare, with most arising from the soft tissues of the presacral area or from the regions dorsal to the sacrum. In even rarer circumstances, the tumor may arise within the sacral canal, likely as a result of ependymal cells of the extradural filum terminale. Because of bone erosion caused by extension of the tumor into the pelvis or dorsal to the sacrum, a truly intraspinal extradural ependymoma in this region has until now never been clearly demonstrated. The authors present a patient with a myxopapillary ependymoma arising from the filum terminale externa in which there was no involvement of the intradural filum or extension outside the sacral canal. A review of the literature is presented, with emphasis on the pathogenesis and clinical management of these rare tumors. PMID- 11012070 TI - Traumatic intratumoral hemorrhage as the presenting symptom of a spinal neurinoma. Case report. AB - Intratumoral hemorrhage as the presenting symptom of spinal tumors is rare. The authors describe a patient who presented with rapidly progressing paraplegia 24 hours after sustaining a minor traumatic injury of the thoracic spine. Radiological evaluation demonstrated a low-thoracic intradural tumor that was resected and found to be a neurinoma in which severe intra- and peritumoral hemorrhage was revealed. The radiological, surgical, and pathological findings are presented and discussed. PMID- 11012071 TI - A rare complication of hardware failure in neurostimulation. Report of two cases. AB - The authors report on a rare complication of neurostimulation. Two patients presented with a skin rash after undergoing neurostimulator implantation, and the implants were found to have faulty electrical insulation. The rash was centered over the source of current leak and disappeared when the problem was corrected. PMID- 11012072 TI - The "open book" technique for preparation of the lumbar transverse process for posterolateral fusion. AB - A new technique is reported for preparation of the recipient graft bed for posterolateral intertransverse process fusion of the lumbar spine. The dorsal surface of each transverse process is reflected open like the pages of a book. This increases the surface area of cancellous bone in the recipient bed, thereby promoting better contact with the grafted bone. This maneuver also preserves much of the periosteal blood supply to the dorsal aspect of the transverse process and much of the cortical bone that would normally be drilled away during decortication. The technical details are described. PMID- 11012073 TI - Vertebral erosion and a ligamentum flavum cyst. Case illustration. PMID- 11012074 TI - Spinal nerve root repair after brachial plexus injury. PMID- 11012075 TI - Interbody cages. PMID- 11012076 TI - The use of methylprednisolone. PMID- 11012077 TI - Diffuse idiopathic skeletal hyperostosis. PMID- 11012078 TI - Endoscopic sympathectomy. PMID- 11012079 TI - DNases and apoptosis. AB - Here we review the different apoptotic DNases. From a functional point of view, DNases implicated in apoptosis may be classified into three groups: the Ca2+/Mg2+ endonucleases, the Mg2+-endonucleases, and the cation-independent endonucleases. The first group includes DNase I which has no specificity for the linker region, DNase gamma which has some homology with DNase I, and other DNases which cleave DNA in the linker region. Both DNase I and DNase gamma have been cloned. The other nucleases of this category have dispersed molecular weights. Their sequences are unknown and it is difficult to determine their role(s) in apoptosis. It seems that different pathways are present and that these nucleases may be activated either by caspases or serine proteases. The caspase 3 activated DNase (CAD, CPAN, or DFF40) belongs to the Mg2+-dependent endonucleases. DNase II belongs to the third group of acid endonucleases or cation-independent DNases. We have shown the involvement of DNase II in lens cell differentiation. Recently, the molecular structure of two different enzymes has been elucidated, one of which has a signal peptide and appears to be secreted. The other, called L-DNase II, is an intracellular protein having two enzymatic activities; in its native form, it is an anti-protease, and after posttranslational modification, it becomes a nuclease. PMID- 11012080 TI - Molecular correlates of the action of bis(ethyl)polyamines in breast cancer cell growth inhibition and apoptosis. AB - Polyamines are known to be involved in cell growth regulation in breast cancer. To evaluate the efficacy of bis(ethyl)polyamine analogs for breast cancer therapy and to understand their mechanism of action we measured the effects of a series of polyamine analogs on cell growth, activities of enzymes involved in polyamine metabolism, intracellular polyamine levels, and the uptake of putrescine and spermidine using MCF-7 breast cancer cells. The IC50 values for cell growth inhibition of three of the compounds, N1,N12-bis(ethyl)spermine, N1,N11 bis(ethyl)norspermine, and N1,N14-bis(ethyl)homospermine, were in the range of 1 2 microM. Another group of three compounds showed antiproliferative activity at about 5 microM level. These compounds are also capable of suppressing colony formation in soft agar assay and inducing apoptosis of MCF-7 cells. The highly effective growth inhibitory agents altered the activity of polyamine biosynthetic and catabolic enzymes and down-regulated the transport of natural polyamines, although each compound produced a unique pattern of alterations in these parameters. HPLC analysis showed that cellular uptake of bis(ethyl)polyamines was highest for bis(ethyl)spermine. We also analyzed polyamine analog conformations and their binding to DNA minor or major grooves by molecular modelling and molecular dynamics simulations. Results of these analyses indicate that tetramine analogs fit well in the minor groove of DNA whereas, larger compounds extend out of the minor groove. Although major groove binding was also possible for the short tetramine analogs, this interaction led to a predominantly bent conformation. Our studies show growth inhibitory activities of several potentially important analogs on breast cancer cells and indicate that multiple sites are involved in the mechanism of action of these analogs. While the activity of an analog may depend on the sum of these different effects, molecular modelling studies indicate a correlation between antiproliferative activity and stable interactions of the analogs with major or minor grooves of DNA. PMID- 11012082 TI - Disruption of LT-antigen/p53 complex by heat treatment correlates with inhibition of DNA synthesis during transforming infection with SV40. AB - Transforming infection of Go/G1-arrested primary mouse kidney cell cultures with simian virus 40 (SV40) induces cells to re-enter the S-phase of the cell cycle. In Go-arrested cells, no p53 is detected, whereas in cells induced to proliferate by infection, a gradual accumulation of p53 complexed to SV40 large T-antigen is observed in the nucleus. Heat treatment of actively proliferating SV40-infected cells leads to inhibition of DNA synthesis and growth arrest. To determine the fate of p53 after heat treatment, proliferating infected cells were exposed to mild heat (42.5 degrees C) for increasing lengths of time. The results presented here show that after ninety minutes of treatment, the arrest of DNA synthesis by heat correlates with the disruption of the p53/LT-antigen complex. Longer treatments induce, in addition, a reduction in the solubility of p53, which was recovered tightly associated with the nuclear fraction. This contrasted with large T-antigen, whose solubility remained unaffected by heat treatment. Although the total amount of p53 in the nucleus remained constant, as shown by immunoblot analyses, p53 was no longer detectable after immunoprecipitation or by immunofluorescent staining techniques. These results suggest that heat treatment had either induced conformational changes in its antigenic sites, or had sequestered the sites through aggregation or binding to insoluble nuclear components. PMID- 11012081 TI - Stress-relaxation and contraction of a collagen matrix induces expression of TGF beta and triggers apoptosis in dermal fibroblasts. AB - Extracellular matrix serves as a scaffold for cells and can also regulate gene expression and ultimately cell behaviour. In this study, we compared the effects of three forms of type I collagen matrix, which differed only in their mechanical properties, and plastic on the expression of transforming growth factor-beta1 (TGF-beta1), matrix metalloproteinase-1 (collagenase), and type I collagen and on the growth and survival of human dermal fibroblasts. These effects were correlated with alterations in cell morphology and organization of intracellular actin. Cells in detached or stress-relaxed matrices were spherical, lacked stress fibres, and showed increased TGF-beta1 mRNA compared to the cells in anchored collagen matrices or on plastic, which were polygonal or bipolar and formed stress fibres. The levels of TGF-beta measured by bioassay were higher in detached and stress-relaxed collagen matrices, than in anchored collagen matrices. Cells on plastic contained little or no immunoreactive TGF-beta, while most cells in collagen matrices were stained. The levels of collagenase mRNA were significantly higher in all the collagen matrix cultures compared to those on plastic, but there were no statistically significant differences between them. Levels of mRNA for procollagen type I were not significantly affected by culture in the collagen matrices. Apoptotic fibroblasts were detected by the TUNEL assay in detached (5.7%) and to a lesser extent in stress-relaxed (2.2%) matrices, but none were observed in anchored collagen matrices or on plastic. These results show that alterations in the mechanical properties of matrix can induce the expression of TGF-beta and trigger apoptosis in dermal fibroblasts. They further suggest that inability to reorganize this matrix could be responsible for the maintenance of the fibroproliferative phenotype associated with fibroblasts in hypertrophic scarring. PMID- 11012083 TI - Effect of pineal indoles on activities of the antioxidant defense enzymes superoxide dismutase, catalase, and glutathione reductase, and levels of reduced and oxidized glutathione in rat tissues. AB - Male Sprague-Dawley rats were randomly divided into four groups. Two of the groups received a single intraperitoneal injection of melatonin and 5 methoxytryptamine (5 mg/kg body weight), respectively, at 9 PM. One group received an intraperitoneal injection of 5-methoxytryptophol (5 mg/kg body weight) at 9 AM. The remaining group received alcoholic saline (vehicle) and served as the control. All rats were sacrificed 90 min after injection and the livers, kidneys, and brains were dissected. The activities of superoxide dismutase, catalase, and glutathione reductase in the organs were measured. It was found that both melatonin and 5-methoxytryptamine were approximately equipotent in enhancing the activities of superoxide dismutase and glutathione reductase in the kidney and liver, while 5-methoxytryptophol displayed a weaker effect. Both melatonin and 5-methoxytryptamine augmented the level of reduced glutathione in the kidney and liver, while 5-methoxytryptophol did so only in the kidney. All three pineal indoles increased the activity of superoxide dismutase and lowered the ratio of oxidized to reduced glutathione in the brain. PMID- 11012084 TI - Identification and characterization of gelatin-cleavage activities in the apically located extracellular matrix of the sea urchin embryo. AB - We have identified and partially characterized several gelatinase activities associated with the sea urchin extraembryonic matrix, the hyaline layer. A previously identified 41-kDa collagenase/gelatinase activity was generally not found to be associated with isolated hyaline layers but was dissociated from the surface of 1-h-old embryos in the absence of Ca2+ and Mg2+. While hyaline layers, freshly prepared from 1-h-old embryos, were devoid of any associated gelatinase activities, upon storage at 4 degrees C for 4 days, a number of gelatin-cleavage activities appeared. Comparative analysis of these activities with the 41-kDa collagenase/gelatinase revealed that all species were inhibited by ethylenediamine tetraacetic acid but were refractory to inhibition with the serine protease inhibitors, phenylmethyl sulfonyl fluoride and benzamidine. In contrast, the largely Zn2+ specific chelator 1,10-phenanthroline had markedly different effects on the gelatinase activities. While several of the storage induced, hyaline-layer-associated gelatinase activities were inhibited, the 41 kDa collagenase/gelatinase was refractory to inhibition as was a second gelatinase species with an apparent molecular mass of 45 kDa. We also examined the effects of a series of divalent metal ions on the gelatin-cleavage activities. In both qualitative and quantitative assays, Ca2+ was the most effective activator while Mn2+, Cu2+, Cd2+, and Zn2+ were all inhibitory. In contrast, Mg2+ had a minimal inhibitory effect on storage-induced gelatinase activities but significantly inhibited the 41-kDa collagenase/gelatinase. These results identify several distinct gelatin-cleavage activities associated with the sea urchin extraembryonic hyaline layer and point to diversity in the biochemical nature of these species. PMID- 11012085 TI - Lectins from bulbs of the Chinese daffodil Narcissus tazetta (family Amaryllidaceae). AB - The isolation of three lectins with similar N-terminal amino acid sequences from the bulbs of the Chinese daffodil Narcissus tazetta was achieved. The isolation protocol involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on mannose-agarose, and fast protein liquid chromatography-gel filtration on Superose 12. The lectins were all adsorbed on mannose-agarose and demonstrated a single band with a molecular weight of 13 kDa in SDS polyacrylamide gel electrophoresis and a single 26 kDa peak in gel filtration, indicating that they were mannose-binding, dimeric proteins. The lectins differed in hemagglutinating activity, with the magnitude of the activity correlating with the ionic strength of the buffer required to elute the lectin from the DEAE cellulose column. The bulb lectin did not exert potent cytotoxicity against cancer cell lines or fetal bovine lung cells but inhibited syncytium formation in, and reinstated viability of, fetal bovine lung cells infected with bovine immunodeficiency virus. PMID- 11012086 TI - Divergent responses of ras-transfected and non-ras-transfected human keratinocytes to extracellular calcium. AB - Raising extracellular calcium (Ca(o)) induces terminal differentiation in cultured epidermal keratinocytes. The introduction of the ras oncogene into keratinocytes results in resistance to Ca(o)-mediated differentiation. To understand the signaling mechanism involved, we examined the Ca(o)-induced formation of inositol triphosphate (IP3) and changes in intracellular Ca2+ (Ca(i)) concentration in non-ras-transfected and ras-transfected HaCaT lines of human keratinocytes. When switched from 0.05- to 1.5-mM Ca(o) medium, the non-ras HaCaT line showed a rapid twofold increase in IP3 formation, whereas the IP3 level in the ras-transfected I-7 line was slightly affected. G-protein-coupled activation of phospholipase was intact in both lines, as evidenced by the generation of similar amounts of IP3 in response to addition of bradykinin or guanosine 5'-[gamma-thio]-triphosphate. Addition of 1.0 mM Ca(o) evoked similar Ca(i) responses in both non-ras- and ras-transfected cells: a transient elevation, followed by a sustained lower plateau. However, the two lines differed in their later responses: after being maintained in 1.0 mM Ca2+ for 24 h, the Ca(i) level was significantly lower in ras-transfected cells than in non-ras transfected HaCaT cells. The Ca(o)-induced increase in Ca(i) in both lines was inhibited by the Ca2+ entry blocker SK&F 96365 or depolarization in high K+ bathing solution, demonstrating its dependence of calcium influx. The results suggest fundamental differences in the early signal that are generated in response to an increase in Ca(o) in ras-transfected keratinocytes, with the absence of a Ca(o)-induced rise in IP3--a signaling pathway defect that may play a role in the differentiation block the cells exhibit. In addition, the inability of ras-transfected cells to sustain a prolonged Ca(i) plateau may also contribute to their inability to differentiate in response to the Ca(o) signal. PMID- 11012087 TI - The ZF87/MAZ transcription factor functions as a growth suppressor in fibroblasts. AB - ZF87/MAZ is a zinc finger transcription factor that activates expression of tissue-specific genes and represses expression of the c-myc proto-oncogene. Infection of NIH3T3 fibroblasts with a retrovirus expressing ZF87/MAZ leads to a significant reduction in G418-resistant colonies, compared to cells infected with a retroviral control. Further, only a small fraction of the G418-resistant colonies express ZF87/MAZ. When the ZF87/MAZ-expressing colonies are expanded, they demonstrate a slow growth phenotype, a delayed transit through G1 phase and a decrease in endogenous c-myc gene expression and cyclin A and cyclin E protein levels. Consistent with a partial G1 arrest, the ZF87/MAZ-expressing cells show a reduced sensitivity to the S phase specific chemotherapeutic agent camptothecin. These data indicate that ZF87/MAZ is a growth suppressor protein in nontransformed cells, in part, by affecting the levels of key cell cycle regulatory proteins. PMID- 11012088 TI - Reversible changes in size of cell nuclei isolated from Amoeba proteus: role of the cytoskeleton. AB - Micrurgically isolated interphasal nuclei of Amoeba proteus, which preserve F actin cytoskeletal shells on their surface, shrink after perfusion with imidazole buffer without ATP, and expand to about 200% of their cross-sectional area upon addition of pyrophosphate. These changes in size may be reproduced several times with the same nucleus. The shrunken nuclei are insensitive to the osmotic effects of sugars and distilled water, whereas the expanded ones react only to the distilled water, showing further swelling. The shrinking-expansion cycles are partially inhibited by cytochalasins. They are attributed to the state of actomyosin complex in the perinuclear cytoskeleton, which is supposed to be in the rigor state in the imidazole buffer without ATP, and to dissociate in the presence of pyrophosphate. Inflow of external medium to the nuclei during dissociation of the myosin from the perinuclear F-actin may be due to colloidal osmosis depending on other macromolecular components of the karyoplasm. PMID- 11012089 TI - Kidney proximal tubule cells: epithelial cells without EGTA-extractable annexins? AB - The expression and the subcellular localizations of annexins I, II, IV, VI, and XIII in renal epithelial cells were investigated, using immunological techniques with specific monoclonal antibodies. Upon performing Western blotting experiments, no annexins VI and XIII were detected in kidney, whereas annexins I, II, and IV were. Immunofluorescence labelling procedure performed on thin frozen renal sections showed the presence of these three annexins along the plasma membrane of the collecting duct cells with a restricted expression of annexin I at principal cells. Annexin I was also found present in some glomerular cells. None of these annexins, however, were detected in the proximal tubular cells upon performing immunofluorescence labelling and electrophoretic analysis on an EGTA (ethylenebis(oxyethylenenitrilo)tetraacetic acid)-extractable annexin fraction prepared from freshly isolated cells. This is the first time a mammalian epithelial cell has been found to express non-typical annexin (at least partly solubilized with EGTA). However, when these cells were grown in primary culture, they were found to express annexins I, II, IV, and V. As well as being located along the basolateral membrane, annexins I and II are also present on vesicles, which suggests that these annexins may be involved in vesicular traffic under cell culture conditions. PMID- 11012090 TI - Contractile elements of Lemna trisulca L. glycerinated cell models during chloroplast translocations. AB - Electron microscopy of Lemna glycerinated cell models depicts contractile elements during chloroplast translocations. One contractile element, the thin ectoplasmic layer, is < or = 0.4 microm thick, pressed against plasma membrane cell wall. Thin ectoplasmic layer contains numerous oriented filaments and some appear to be actin and myosin. Another contractile element is the outer chloroplast membrane which envelops each chloroplast and joins or fuses with the thin ectoplasmic layer. Chloroplast interconnections are formed between two or more chloroplasts by outer chloroplast membranes; they enhance chloroplast communications, translocations, and molecular exchanges. PMID- 11012091 TI - Expression of heat shock protein 47(Hsp47) mRNA levels in rabbit connective tissues during the response to injury and in pregnancy. AB - Hsp47 (also termed "colligin") is a 47 kDa protein that is localized in the ER and cis-Golgi vesicles of fibrocytes, chondrocytes, and other collagen-secreting cells. Under stress conditions, Hsp47 expression is upregulated as part of the heat shock/stress response that mitigates cell damage from noxious stimuli such as elevated temperature, heavy metals, and oxidative stress. Under non-stress conditions, Hsp47 functions as a collagen-specific molecular chaperone that facilitates intracellular procollagen polypeptide synthesis, and triple helix assembly in connective tissues. Previously it has been shown that levels of collagen-specific gene expression are significantly altered in ligaments, menisci, and other connective tissues of the rabbit following surgically induced injuries (increased), and during pregnancy (decreased). The present study was undertaken to determine whether expression of mRNA for the Hsp47 collagen-binding stress protein was also influenced in these experimental models. Since no sequence information was available on the rabbit Hsp47 gene, a partial cDNA for rabbit Hsp47 was first isolated and cloned using reverse transcriptase PCR (RT PCR) with degenerate oligonucleotide primers. Rabbit Hsp47 sequence-specific primers then designed enabled analysis of Hsp47 mRNA expression in rabbit connective tissues using semiquantitative RT-PCR. It was found that Hsp47 expression is affected in a complex, tissue-specific manner by injury and pregnancy. Hsp47 transcript levels were elevated in the medial collateral ligament (MCL) of the rabbit knee following surgical induction of a gap injury. Transection of the anterior cruciate ligament (ACL), which leads to chronic progressive damage to menisci of the rabbit knee joint, was accompanied by an upregulation of Hsp47 expression in the medial and lateral menisci. Hsp47 mRNA levels were depressed during pregnancy in the kidney and ACL of primigravid adolescent rabbits, but were not altered in corneal tissue during pregnancy or in the ACL of skeletally mature multiparous females. The changes in Hsp47 transcript levels within these connective tissues following injury/pregnancy often, but not always, paralleled changes in collagen-specific gene expression. PMID- 11012092 TI - Characterization of the interaction of recombinant apolipoprotein(a) with modified fibrinogen surfaces and fibrin clots. AB - Elevated levels of lipoprotein(a) [Lp(a)] in plasma are a significant risk factor for the development of atherosclerotic disease, a property which may arise from the ability of this lipoprotein to inhibit fibrinolysis. In the present study we have quantitated the binding of recombinant forms of apolipoprotein(a) [17K and 12K r-apo(a); containing 8 and 3 copies, respectively, of the major repeat kringle sequence (kringle IV type 2)] to modified fibrinogen surfaces. Iodinated 17K and 12K r-apo(a) bound to immobilized thrombin-modified fibrinogen (i.e., fibrin) surfaces with similar affinities (Kd approximately 1.2-1.6 microM). The total concentration of binding sites (Bmax) present on the fibrin surface was approximately 4-fold greater for the 12K than for the 17K (Bmax values of 0.81 +/ 0.09 nM, and 0.20 +/- 0.01 nM respectively), suggesting that the total binding capacity on fibrin surfaces is reduced for larger apolipoprotein(a) (apo(a)) species. Interestingly, binding of apo(a) to intact fibrin was not detected as assessed by measurement of intrinsic fluorescence of free apo(a) present in the supernatants of sedimented fibrin clots. In other experiments, the total concentration apo(a) binding sites available on plasmin-modified fibrinogen surfaces was shown to be 13.5-fold higher than the number of sites available on unmodified fibrin surfaces (Bmax values of 2.7 +/- 0.3 nM and 0.20 +/- 0.01 nM respectively) while the affinity of apo(a) for these surfaces was similar. The increase in Bmax was correlated with plasmin-mediated exposure of C-terminal lysines since treatment of plasmin-modified fibrinogen surfaces with carboxypeptidase B produced a significant decrease in total binding signal as detected by ELISA (enzyme linked immunosorbent assay). Taken together, these data suggest that apo(a) binds to fibrin with poor affinity (low microM) and that the total concentration of apo(a) binding sites available on modified-fibrinogen surfaces is affected by both apo(a) isoform size and by the increased availability of C-terminal lysines on plasmin-degraded fibrinogen surfaces. However, the low affinity of apo(a) for fibrin indicates that Lp(a) may inhibit fibrinolysis through a mechanism distinct from binding to fibrin, such as binding to plasminogen. PMID- 11012093 TI - Transforming growth factor beta1 selectively regulates ferritin gene expression in malignant H-ras-transformed fibrosarcoma cell lines. AB - Transforming growth factor beta1 is an important growth regulator in many cell types, usually exerting a negative effect on cellular growth. Inhibition of DNA synthesis and cell proliferation is frequently lost during malignant transformation, and in some cases, tumor cell proliferation is actually stimulated by TGF-beta1. The present study demonstrates a novel link between alterations in TGF-beta1 regulation during malignant conversion, and the expression of ferritin, an important activity involved in a number of biological functions including iron homeostasis and cell-growth control. A series of H-ras transformed mouse 10 T 1/2 cell lines, exhibiting increasing malignant potential, was investigated for possible TGF-beta1-mediated changes in ferritin gene expression. Selective induction of gene expression was observed, since only H-ras transformed cells with malignant potential exhibited marked elevations in ferritin gene expression, in particular, alterations in H-ferritin gene expression. The regulation of H-ferritin gene expression in response to TGF-beta1 did not involve alterations in transcription, but occurred through mechanisms of post-transcriptional stabilization of the H-ferritin mRNA. Additionally, evidence was obtained for a cycloheximide-sensitive regulator of H-ferritin gene expression, since the presence of this protein synthesis inhibitor increased H ferritin message levels, and in combination with TGF-beta1, cooperated in an additive manner to augment H-ferritin gene expression. These results show for the first time that TGF-beta1 can regulate ferritin gene expression in malignant H ras transformed cells, and suggest a mechanism for growth factor stimulation of malignant cells, in which early alterations in the control of H-ferritin gene expression are important. PMID- 11012094 TI - The role of blood tumor marker measurement (using a biochemical index score and c erbB2) in directing chemotherapy in metastatic breast cancer. AB - The role of blood tumor markers in monitoring response in advanced breast cancer is established in endocrine therapy and standard chemotherapy. This study examines marker levels in patients receiving new chemotherapy regimens. Thirty patients were recruited into two multicenter trials in which docetaxel-based regimens were used in 15 patients. The other 15 received doxorubicin-based regimens. Biochemical response calculated from a score using CA15.3, CEA and ESR was compared with UICC response. Marker changes at 2, 4 and 5 months correlated with UICC response at 3, 4(1/2) and 6 months, respectively (p < 0.03). Eleven patients achieved both clinical/radiological and biochemical response at the end of treatment; markers had not yet returned to below cutoffs in seven, suggesting a possible advantage to continue chemotherapy. No patient showed a biochemical response whilst judged clinically/radiologically progressive. Nineteen patients had progressed either clinically/radiologically or biochemically at six months; of these, eight showed progression assessed earlier by markers so that a median of four cycles of chemotherapy could have been saved. Measurements of serum c erbB2 showed a correlation with tissue c-erbB2 staining in the primary tumor (p < 0.003). Among the patients with positive tissue staining, sequential changes in serum c-erbB2 completely paralleled initial response. PMID- 11012095 TI - CCND1 mRNA overexpression is highly related to estrogen receptor positivity but not to proliferative markers in primary breast cancer. AB - To elucidate the role of CCND1 alterations in sporadic breast cancer we investigated the possible link between CCND1 mRNA levels versus estrogen-receptor (ER) status and a proliferation marker, S-phase fraction (SPF), measured by flow cytometry. CCND1 expression was quantified by means of real-time quantitative RT PCR in a well-characterized series of 33 primary breast cancer patients. Eighteen tumors (54.5%) showed CCND1 overexpression ranging from 3.3 to 29.5 times the level observed in normal breast tissue. Seventeen (94.4%) of the 18 cases with CCND1 overexpression were ER-positive compared to seven (46.7%) of the 15 cases with normal CCND1 expression (p=0.0074). CCND1 overexpression was independent of SPF and DNA-ploidy status. These data suggest that the CCND1 gene does not act as an oncogene responsible for more rapid cell proliferation in breast cancer, but could be involved in the regulation of hormone sensitivity associated with ER. PMID- 11012096 TI - Relationship of tumoral hyaluronic acid and cathepsin D contents with histological type of gastric carcinoma. AB - The aim of this work was to evaluate the cytosolic contents of hyaluronic acid (HA) and cathepsin D (CatD) in gastric carcinomas and their possible relationships with the clinicopathological parameters of the tumors. Our study demonstrated a wide variability in the cytosolic levels of HA (mean +/- SEM: 3748 +/- 411 ng/mg protein) and cathepsin D (52 +/- 4 pmol/mg protein) in the tumors of 78 gastric cancer patients. In addition, the tumoral contents of HA and CatD were significantly higher (p<0.005) in diffuse type (HA: 6027 +/- 1099 ng/mg protein; CatD: 75 +/- 13 pmol/mg protein) than in intestinal type (HA: 2735 +/- 242 ng/mg protein; CatD: 42 +/- 3 pmol/mg protein) carcinomas. These data suggest that both markers may contribute to the biological characterization of gastric carcinomas. PMID- 11012097 TI - BTA-TRAK combined with urinary cytology is a reliable urinary indicator of recurrent transitional cell carcinoma (TCC) of the bladder. AB - This study evaluated the diagnostic accuracy of BTA-TRAK in combination with urinary cytology (UC) in the follow-up of patients with a history of transitional cell carcinoma (TCC) of the bladder. The overall sensitivity of BTA-TRAK, UC and the two tests combined for the detection of recurrences was 82.7% (48/58), 84.2% (48/57) and 91.2% (52/57), respectively. BTA and UC showed comparable sensitivity for superficial recurrences (76.7% (33/43) and 78.5% (33/42), respectively) and for invasive recurrences (100% (15/15)); when the two tests were used in combination, the sensitivity for superficial lesions increased to 88% (37/42). BTA-TRAK was more sensitive than UC for G1 recurrences (81.2% (13/16) vs. 68.7% (11/16)), and when the two tests were combined the sensitivity increased to 87.5% (14/16). The sensitivity of the combination was 100% (15/15) for G3 lesions. The differences in urinary BTA-TRAK levels between patients with recurrences and those without evidence of disease were statistically significant (Wilcoxon's test, p<0.05); among patients with recurrences BTA levels were significantly higher in the invasive and poorly differentiated subtypes. In the series of patients studied by us, BTA-TRAK combined with UC was shown to be a non-invasive, accurate test to predict TCC recurrences. Periodic measurement of BTA-TRAK combined with urinary cytology seems to provide additional information for the monitoring of patients treated for TCC; however, due to the presence of false positive and false negative results, this test cannot replace cystoscopy. In a selected group of patients it could, if combined with cytology and ultrasonography and if correctly used and interpreted, orient the timing and indication for cystoscopy. PMID- 11012098 TI - Cholestasis is the main determinant of abnormal CA 19-9 levels in patients with liver cirrhosis. AB - BACKGROUND/AIMS: Altered CA19-9 levels are commonly found in patients with liver cirrhosis though a clear explanation for this finding has not yet been given. The aim of this study was to investigate whether CA19-9 levels might be related to alterations in biochemical parameters and/or to functional impairment in cirrhotic patients with and without hepatocellular carcinoma. METHODS: We studied 126 patients with liver cirrhosis, 60 of whom also had hepatocellular carcinoma. CA19-9 values were related to clinical, biochemical and functional parameters. In half of the patients CA19-9 levels were related to the monoethylglycinexylidide test, which is a dynamic liver function test. RESULTS: In more than half the cases CA19-9 values were above the upper limit. Liver function worsening as assessed by Child-Pugh's score and monoethylglycinexylidide test did not seem to influence the alteration of the marker. By contrast, in univariate analysis CA19 9 correlated with aminotransferases, gamma-glutamyltransferase and alkaline phosphatase. Multivariate analysis showed that besides alkaline phosphatase also the presence of hepatocellular carcinoma might influence the alteration of CA19 9, although the marker was of no use for the diagnosis of liver cancer in patients with altered though not diagnostic alpha-fetoprotein levels. CONCLUSIONS: In our study we confirmed the correlation of CA19-9 levels with cholestasis and cytolysis parameters. Moreover, we found no association between CA19-9 levels and impaired liver function as assessed by means of the Child Pugh's score and the monoethylglycinexylidide test, which is cholestasis independent and explores liver metabolic and clearance activities. The cholestatic picture that characterizes liver cirrhosis might enhance the expression and passage of the marker from the bile to the blood. The addition of CA19-9 assessment is not useful for the diagnosis of hepatocellular carcinoma in patients with non-diagnostic levels of alpha-fetoprotein. Caution should therefore be used when evaluating CA19-9 in cirrhotic patients with cholestasis, since false positive results may occur. PMID- 11012099 TI - C-erbB-2 oncoprotein content in gastric cancer and in adjacent mucosa. AB - The aim of this study was to evaluate, by means of an immunoenzymatic assay, the membranous and cytosolic c-erbB-2 oncoprotein contents in primary tumors and in adjacent mucosa from gastric cancer patients. Fifty-two patients with primary gastric adenocarcinomas were enrolled in this prospective study. c-erbB-2 protein levels were significantly higher in membranous than in cytosolic samples, both in neoplastic tissues (median: 3602 vs 525 NHU/mg protein; p<0.0001) and in adjacent mucosa samples (median: 3174 vs 509 NHU/mg protein; p<0.0001). Nevertheless, there was a significant positive relation between membranous and cytosolic c-erbB 2 protein contents in both neoplastic tissue (p<0.001) and adjacent mucosa (p<0.001) samples. There was no significant difference in the membranous c-erbB-2 protein content between neoplastic tissues and adjacent mucosa samples. However, the cytosolic c-erbB-2 content was significantly higher in neoplastic tissues than in adjacent mucosa (p<0.05). Finally, the results did not show any significant correlations of these oncoprotein contents with patient characteristics, clinicopathologic parameters and overall survival of the study population. PMID- 11012100 TI - Clinical evidence for correlation of insufficient tissue oxygen supply (hypoxia) and tumor-associated proteolysis in squamous cell carcinoma of the head and neck. AB - Hypoxic tumors of patients with squamous cell carcinoma of the head and neck show a consistent trend towards poor treatment outcome. We now report that tumor hypoxia in these patients is correlated with elevated antigen content of the tumor-associated serine protease uPA (urokinase-type plasminogen activator), a marker of tumor cell invasion and metastasis. PMID- 11012101 TI - Towards a "good" death: end-of-life narratives constructed in an intensive care unit. AB - End-of-life decisions regarding the withdrawal and withholding of life supporting technology have become commonplace within intensive care units (ICUs). In this paper, we examine the dialogue between ICU team members and families regarding limitation of treatment as a therapeutic narrative--that is, as a story which frames therapeutic events as well as the critically ill patient's experience in a meaningful and psychologically comforting way for families and health care providers alike. The key themes of these end-of-life narratives are discussed, as well as the qualities that the stories share with other narratives of the same genre. PMID- 11012102 TI - Cultural identity and illness: Fulani views. AB - Cultural identity--who the Fulani think they are--informs thinking on illnesses they suffer. Conversely, illness, so very prevalent in sub-Saharan Africa, provides Fulani with a constant reminder of their distinctive condition in Guinea. How they approach being ill also tells Fulani about themselves. The manner in which Fulani think they are sick expresses their sense of difference from other ethnic groups. Schemas of illness and of collective identity draw deeply from the same well and web of thoughts. Three different approaches of schema theory are used to trace what ties illness to identity. These are 1) the schema as prototype; 2) a connectionist approach associates schemas for illness with other cultural schemas; and 3) a hierarchy of schemas. The hierarchy includes master schemas for ethnic identity, schemas for illness generally and sub-schemas for separate ailments. Schemas orient and provide a framework for the practice of being Fulani--in the sense that Bourdieu would describe practice as the application of practical knowledge. Illnesses above the waist are said to be part of the Fulani condition of belonging in arid climates while they need suffer the humidity of Guinea. Illnesses below the waist are thought to arise when one does not act like a Fulani, especially in matters of food and sex. As individuals disclose or conceal illness, as they discuss illness and the problem of others they reflect standards of Fulani life--being strong of character not necessarily of body, being disciplined, rigorously Moslem, and leaders among lessors. To disregard standards or to suggest one does not care about such standards is shameful and places one out of phase with others and with cultural norms. But to be in step with others and with cultural norms is to have pride in the self and the foundations of Fulani life. PMID- 11012103 TI - False teeth and real suffering: the social course of 'germectomy' in eastern Uganda. AB - The removal of the incipient canine teeth ('germectomy') in small babies is a practice carried out in many parts of eastern Africa. This article describes how 'germectomy' among the Jop'Adhola in Eastern Uganda is an important idiom of distress, referred to as false teeth by English speaking people, and lakijo marach (bad teeth) or gira kwanya (that which is removed) in the local language Dhop'Adhola. Through an analysis of how the notion of false teeth is shaped by macro social forces of war and poverty as well as by negotiations within the local social world, the discussion is taken beyond the question of cultural belief. False teeth as a practice seems to have spread through vast geographical areas within a few decades, but as the example of the Jop'Adhola shows, it has taken a particular social course in eastern Uganda--as it is most likely to also have done everywhere else it has gained a footing. By analyzing its social course we may gain insight into important mediating social processes which may have as much to do with actual health outcome in a particular area as health care per se. PMID- 11012104 TI - Latinas, amniocentesis and the discourse of choice. AB - Little attention has been paid to the impact of the increasingly routine use of fetal diagnosis on how U.S. minority women experience their pregnancies and decide whether to have their fetuses tested. Using narrative analysis, we offer the account of one Latina who, despite considerable turmoil, ultimately accepted an offer of amniocentesis. We describe her reasoning in choosing a course of action. Data from interviews with 147 Latinas who were faced with the same decision are used to contextualize the case study material. We seek to illuminate how a blending of Mexican and European American cultural influences helped shape the woman's experience and define the dilemma she faced when she learned her fetus might be born with a grave or incurable condition because she was ideologically opposed to abortion. PMID- 11012105 TI - Balloon dilation of cor triatriatum dexter in a dog. PMID- 11012106 TI - The case for measuring plasma colloid osmotic pressure. PMID- 11012107 TI - Measurement of plasma colloid osmotic pressure in normal thoroughbred neonatal foals. AB - A normal plasma colloid osmotic pressure (COP) interval was established for foals and compared to values for adult horses. Plasma samples were obtained from 38 Thoroughbred foals that had normal findings on postfoaling examination and 10 healthy Thoroughbred adult horses. Samples were analyzed using a commercially available colloid osmometer. Fifty samples were obtained from 38 foals. Twelve foals had 2 samples taken, 1 during the 1st 24 hours of life and the 2nd between 24 and 72 hours of life. For foals with 2 samples, only 1 randomly selected value was used in group analysis. Total plasma protein, albumin, and globulin concentrations were measured on all samples from foals. The mean measured plasma COP for foals was 18.8 +/- 1.9 mm Hg for the 38 samples analyzed. Measured plasma COP did not differ significantly over the time period examined for either the 12 paired samples (P = .13) or with regression analysis of the 38 samples (P = .13). Calculation of mean COP, based on previously published quadratic equations using total protein, albumin, and globulin concentrations, underestimated mean measured foal COP values except for when total protein measured by refractometer was used in the Landis-Pappenheimer equation. In conclusion, the plasma COP interval (95% CI: 15.0 mm Hg, 22.6 mm Hg) obtained for healthy foals in this study was found to be lower than that of healthy adult Thoroughbreds (20.6 +/- 0.7 mm Hg, P = .006). PMID- 11012108 TI - Treatment of canine hemangiosarcoma: 2000 and beyond. AB - Canine hemangiosarcoma (HSA) is an aggressive and malignant neoplasia with a grave prognosis. Surgery and chemotherapy have limited success in prolonging survival times and increasing quality of life in dogs with HSA. Advances in medical oncology are resulting in increased survival rates and a better quality of life for veterinary cancer patients. An understanding of mechanisms of metastasis has led to the development of new treatments designed to delay or inhibit tumor spread. Promising new treatment options include novel delivery systems (inhalation or intracavitary chemotherapy); use of immunomodulators such as liposome-encapsulated muramyl tripeptide-phosphatidylethanolamine; antimetastatic agents such as inhibitors of angiogenesis (interferons, thalidomide), matrix metalloproteinase inhibitors, and minocycline; dietary modifications; and gene therapy. Inhibitors of angiogenesis seem to be safe and, unlike conventional chemotherapy, do not induce drug resistance. Although many of the newer approaches are still under development and review, the use of multimodality therapy incorporating innovative treatment modalities may offer the best therapeutic option for dogs affected with HSA. PMID- 11012109 TI - Prognostic factors in dogs with urinary bladder carcinoma. AB - Medical records and biopsy specimens were retrospectively reviewed from 25 dogs diagnosed with unresectable urinary bladder carcinoma and treated with chemotherapy. Our intention was to identify clinical, histologic, and immunohistochemical indicators of prognosis. Immunohistochemical stains for P glycoprotein, glutathione-S-transferase pi, and factor VIII-related antigen were applied to archived tissue. There were more spayed female dogs than castrated male dogs (76% versus 24%). Transitional cell carcinoma was the most common tumor (88%, n = 22), followed by undifferentiated carcinoma (8%, n = 2) and squamous cell carcinoma (4%, n = 1). Overall median survival was 251 days. Histologic diagnosis and immunohistochemical characteristics did not correlate with prognosis. Spayed females survived significantly longer than castrated males (358 days versus 145 days, P = .042). Dogs that received either doxorubicin or mitoxantrone in addition to a platinum-based chemotherapeutic (either cisplatin or carboplatin) lived significantly longer than those that received only a platinum compound (358 days versus 132 days, P = .042). PMID- 11012110 TI - Primary myelodysplastic syndromes of dogs: a report of 12 cases. AB - Clinical bone marrow specimens submitted to the University of Minnesota's Veterinary Teaching Hospital Cytology Service over a 3-year period were evaluated for the presence of myelodysplastic features. Of 220 bone marrow specimens examined, 30 contained dysplastic features. Twenty-seven of these dogs were evaluated further. Twelve were categorized as primary myelodysplastic syndromes, and 15 were categorized as secondary myelodysplastic syndromes. Of the primary myelodysplastic syndromes, 4 were subcategorized as refractory anemia and 8 were categorized as myelodysplasia. Primary refractory anemia was characterized by nonregenerative anemia without leukopenia or thrombocytopenia and with prolonged survival. Primary myelodysplasia was characterized by pancytopenia, greater than 5% myeloblasts in bone marrow, dysplastic features in all bone marrow cell lines, and short survival time. Results of this study indicate that differentiating primary refractory anemia from primary myelodysplasia has both therapeutic and prognostic significance. Dogs with primary refractory anemia tend to have prolonged survival and respond to erythropoietin treatment, whereas dogs with primary myelodysplasia have short survival and do not respond to standard treatments. PMID- 11012111 TI - Cisplatin and doxorubicin combination chemotherapy for the treatment of canine osteosarcoma: a pilot study. AB - Sixteen dogs with histologically confirmed appendicular osteosarcoma were treated by amputation followed by cisplatin and doxorubicin chemotherapy. All dogs began chemotherapy within 24 hours of surgery. Cisplatin was administered at 50 mg/m2 intravenously (IV) concurrent with saline-induced diuresis. Doxorubicin was administered 24 hours later at 15 mg/m2 as a slow IV bolus. This protocol was given on a 21-day cycle for 4 cycles. No dose delays were required, but dose reduction of doxorubicin was required in 2 dogs because of neutropenia. Thoracic radiography was performed every 2 months after completion of therapy to monitor for metastatic disease. Two dogs were still alive and free from disease at the time of last contact (24 and 75 months, respectively). Postmortem examinations were performed on 13 of the 14 dogs that died. Eight of these dogs were euthanized because of metastatic osteosarcoma. Of the remaining 5 dogs, euthanasia was performed because of complications of idiopathic megaesophagus (n = 1), arthritis (n = 2), and hemangiosarcoma (n = 2). The median disease-free interval and survival times were 15.7 and 18 months, respectively. When compared to a historical group of 36 dogs with appendicular osteosarcoma treated with surgery and 4 doses of cisplatin. both disease-free interval and overall survival were significantly longer in the study population (P < .015 and P < .007, respectively). PMID- 11012112 TI - Thiopurine methyltransferase in red blood cells of dogs, cats, and horses. AB - Our objective was to determine if thiopurine methyltransferase (TPMT), the enzyme important in the metabolism of azathioprine in human beings, is detectable in red blood cell lysates (RBCL) of healthy dogs, cats, and horses. Values for TPMT activity were determined from blood collected from 20 healthy dogs, cats, and horses. The TPMT activity in each animal's RBCL was determined using a radioenzymatic end point involving TPMT-facilitated metabolism of 6 mercaptopurine to 6-methylmercaptopurine (6-MMP). One unit of TPMT activity represents the formation of 1 nmol of 6-MMP per milliliter of packed red blood cells per hour of incubation at 37 degrees C. TPMT activity in RBCL was detectable in all species, with mean RBC values +/- standard deviation of 17.9 +/ 3.79 U/mL in dogs; 2.76 +/- 0.70 U/mL in cats; and 2.185 +/- 0.36 U/mL in horses. Values for TPMT in the 3 species were significantly (P < .05) different from one another. TPMT values for dogs were significantly higher than the other species, and TPMT values for cats were significantly higher than those for horses. We conclude that RBCL TPMT values are measurable in dogs. cats, and horses and that dogs have higher values than cats or horses. These findings are consistent with the lower tolerance for azathioprine in cats as compared with dogs. It remains to be determined whether RBCL TPMT values in these species correlate with TPMT activity in the liver, where most of the metabolization of azathioprine is believed to occur. PMID- 11012113 TI - Serum alhpa 1-acid glycoprotein concentrations in healthy and tumor-bearing cats. AB - The purpose of this study was to evaluate alpha 1-acid glycoprotein (AGP) concentrations in tumor-bearing and healthy cats. The hypothesis of the present study was that AGP concentrations would be significantly increased in tumor bearing cats. Serum from 51 healthy and 97 tumor-bearing, client-owned cats was harvested at the time of presentation and stored at -80 degrees C until assayed. Cats with measurable, histologically confirmed malignancies, and healthy cats of similar ages were included. Serum was assayed for AGP concentration by using a radial immunodiffusion method. AGP concentrations were significantly (P = .0051) higher in tumor-bearing (763 +/- 595 microg/mL; mean +/- SD) when compared to healthy cats (501 +/- 377 microg/mL; mean +/- SD). Of the tumor-bearing cats, 35 had carcinomas, 33 had sarcomas, and 26 had discrete, round cell tumors. AGP concentrations were 645 +/- 62 microg/mL, 660 +/- 540 microg/mL, and 967 +/- 860 microg/mL, respectively, and there were no significant differences among the groups. PMID- 11012114 TI - Plasma homocysteine, B vitamins, and amino acid concentrations in cats with cardiomyopathy and arterial thromboembolism. AB - Arterial thromboembolism (ATE) is a common complication of cats with cardiomyopathy (CM), but little is known about the pathophysiology of ATE. In people, high plasma concentrations of homocysteine and low B vitamin concentrations are risk factors for peripheral vascular disease. In addition, low plasma arginine concentrations have been linked to endothelial dysfunction. The purpose of this study was to compare concentrations of homocysteine, B vitamins, and amino acids in plasma of normal cats to those of cats with CM and ATE. Plasma concentrations of homocysteine, vitamin B6, vitamin B12, folate, and amino acids were measured in 29 healthy cats, 27 cats with CM alone, and 28 cats with both CM and ATE. No differences were found between groups in homocysteine or folate. Mean vitamin B12 concentration (mean +/- standard deviation) was lower in cats with ATE (866 +/- 367 pg/mL) and cats with CM (939 +/- 389 pg/mL) compared with healthy controls (1,650 +/- 700 pg/mL; P < .001). Mean vitamin B6 concentration was lower in cats with ATE (3,247 +/- 1.215 pmol/mL) and cats with CM (3,200 +/- 906 pmol/mL) compared with healthy control animals (4,380 +/- 1,302 pmol/mL; P = .005). Plasma arginine concentrations were lower in cats with ATE (75 +/- 33 nmol/mL) compared with cats with CM (106 +/- 25 nmol/mL) and healthy control animals (96 +/- 25 nmol/ mL; P < .001). Vitamin B12 concentration was significantly correlated with left atrial size. We interpret the results of this study to suggest that vitamin B12 and arginine may play a role in CM and ATE of cats. PMID- 11012115 TI - Clinical, echocardiographic, and neurohormonal effects of a sodium-restricted diet in dogs with heart failure. AB - The use of low-sodium diets in dogs with heart failure is common practice, but randomized, double-blind studies have not been conducted to examine the benefits or problems with this approach. The purpose of this study was to determine the effects of a low-sodium diet on clinical, echocardiographic, and neurohormonal parameters in dogs with heart failure. Dogs with stable chronic heart failure were fed exclusively a low-sodium (LS) and a moderate-sodium (MS) diet for 4 weeks each in a randomized, double-blind, crossover design. At days 0, 28, and 56, echocardiography and thoracic radiography were performed, and blood was analyzed for electrolytes and neurohormones. Fourteen dogs completed the study (9 with chronic valvular disease and 5 with dilated cardiomyopathy). Electrolyte abnormalities were common during the study, and serum sodium and chloride concentrations decreased significantly on the LS diet. Neurohormones did not change significantly between diet groups. Maximum left atrial (P = .05) and standard left atrial (P = .09) size decreased on the LS diet. For dogs with chronic valvular disease, vertebral heart score (P = .05), left ventricular internal dimension in diastole (P = .006) and systole (P = .02), standard left atrial dimension (P = .03), maximum left atrial dimension (P = .02), end diastolic volume index (P = .02), and end-systolic volume index (P = .04) decreased significantly on the LS diet compared to the MS diet. Although analysis of these data suggests some benefits of a low-sodium diet, future studies with improved study design are needed to further evaluate the advantages and disadvantages of sodium restriction in dogs with heart failure. PMID- 11012116 TI - Cats surviving natural infection with Cytauxzoon felis: 18 cases (1997-1998). AB - Eighteen cats surviving natural infection with Cytauxzoon felis were identified. All cats came from a limited geographic area in northwestern Arkansas and northeastern Oklahoma. Clinical signs in most cats were similar to those described for cytauxzoonosis; however, 4 cats were asymptomatic. All cases were initially diagnosed by microscopic identification of signet ring-shaped piroplasms in erythrocytes of peripheral blood smears. Four of 4 cats tested had detectable serum antibodies to C felis. Four different cats were positive by polymerase chain reaction (PCR). Partial sequencing of the PCR product from 1 cat revealed >99% homology with the reported sequence of C felis. Repeated examination of blood smears from 12 cats revealed that the erythroparasitemia was generally persistent for the duration of follow-up (3-154 days). Survival did not seem dependent on treatment, as only 1 cat was treated with a drug with potential antiprotozoal activity (imidocarb dipropionate), and 4 cats received no treatment. The findings of this study may indicate the existence of a less virulent strain of C felis. PMID- 11012117 TI - Effects of enalapril versus placebo as a treatment for canine idiopathic glomerulonephritis. AB - A blinded, multicenter, prospective clinical trial assessed the effects of enalapril (EN) versus standard care in dogs with naturally occurring, idiopathic glomerulonephritis (GN). Twenty-nine adult dogs with membranous (n = 16) and membranoproliferative (n = 13) GN were studied. Dogs were randomly assigned to receive either EN (0.5 mg/kg PO q12-24h; n = 16) or placebo (n = 14) for 6 months (1 dog was treated first with the placebo and then with EN). All dogs were treated with low-dose aspirin (0.5-5 mg/kg PO q12-24h) and fed a commercial diet. At baseline, serum creatinine (SrCr), systolic blood pressure (SBP), and glomerular histologic grade were not different between groups, but the urine protein/creatinine ratio (UP/C) was greater in the EN group compared with the placebo group (8.7 +/- 4.4 versus 4.7 +/- 2.3). After 6 months of treatment, the change in UP/C from baseline was significantly different between groups (EN = 4.2 +/- 1.4 versus 1.9 +/- 0.9 in the placebo group). When data were adjusted for changes in SrCr (SrCr X UP/C) a similar significant reduction was noted ( 2.2 +/- 15.2 versus 8.4 +/- 10.1). The change in SBP after 6 months of treatment also was significantly different between groups (EN = -12.8 +/- 27.3 versus 5.9 +/- 21.5 mm Hg in the placebo group). Response to treatment was categorized as improvement (assigned a value of 2), no progression (assigned a value of 1), and progression (assigned a value of 0). Response was significantly better in the EN group (1.4 +/- 0.8) compared with the placebo group (0.3 +/- 0.5). These results suggest that EN treatment is beneficial in dogs with naturally occurring idiopathic GN. PMID- 11012118 TI - Quantitative bacterial cultures and cytological examination of bronchoalveolar lavage specimens in dogs. AB - Cytology and quantitative bacterial cultures of lower respiratory tract secretions are widely used in human medicine to differentiate airway infection from simple bacterial colonization. A retrospective study was conducted to determine the usefulness of quantitative aerobic cultures and Gram stain intracellular bacteria counts from bronchoalveolar lavage (BAL) specimens in dogs in diagnosing lower respiratory tract infection (LRTI) and to determine whether chronic bronchitis is associated with marked bacterial growth in dogs. The threshold determined to define clinically relevant bacterial growth was 1.7 x 10(3) colony-forming units per milliliter of BAL fluid. We used this threshold and found that diagnostic sensitivity and specificity were 86% and 100%, respectively. With a threshold for infection of >2 intracellular bacteria observed in any of 50 fields, microscopic examination of Gram stain BAL preparations had a sensitivity of 71% and a specificity of 97% in establishing LRTI. There was a high correlation between bacterial morphology on BAL Gram stain and bacterial cultures. Combining the results of intracellular bacteria counts from the BAL Gram stain with those from the quantitative cultures, the sensitivity in diagnosing LRTI was 87% and the specificity was 97%. BAL quantitative cultures as well as quantitating intracellular bacteria on Gram stain BAL cytology were revealed to be useful in identifying LRTI in dogs. Chronic bronchitis does not appear to be associated with marked bacterial growth in dogs. PMID- 11012119 TI - Reduced serum insulin-like growth factor (IGF) and IGF-binding protein-3 concentrations in two deerhounds with congenital portosystemic shunts. PMID- 11012120 TI - Pulmonary alveolar proteinosis in a dog. PMID- 11012121 TI - Demonstration of thiopurine methyltransferase activity in the erythrocytes of cats. AB - Azathioprine is a purine analogue used as an immunosuppressive and immunomodulator agent in various mammals, including cats. Several adverse reactions have been reported and have limited the use of the drug in the cat. Adverse reactions to azathioprine in humans have been correlated with reduced activity of thiopurine methyltransferase (TPMT) in erythrocytes. The purpose of this preliminary study was to determine if cats have TPMT activity in their erythrocytes and to compare the values obtained with the normal range for humans and the normal range for dogs in a preliminary report. Activity of the enzyme was measured in blood samples drawn from 41 cats. Blood also was taken from 5 dogs. The mean erythrocyte TPMT activity in the cats was 2.4 +/- 0.4 nmol (range, 1.2 3.9 nmol) per hour per milliliter of red blood cells (U/mL RBC) or 2-8 nmol per hour per gram of hemoglobin (U/g Hb). This range was far lower than the normal human range (8-15 U/mL RBC; 16-33 U/g Hb) and was of monopolar distribution. This observation apparently precludes any diagnostic purpose in assaying erythrocyte TPMT in this species. Erythrocyte TPMT activity in the 5 dogs ranged from 5.5 to 13.1 U/mL RBC (11-27 U/g Hb), which was comparable with normal and carrier ranges for humans, but proof of TPMT genetic polymorphism in either species will require genotyping studies. PMID- 11012122 TI - Enabling the quality in care. PMID- 11012123 TI - Unilateral neonatal testicular torsion. AB - Testicular torsion in the neonatal period is an unusual finding. Because of the high morbidity associated with this condition, early recognition and appropriate management are imperative. Testicular torsion may be unilateral or bilateral and may occur prenatally or in the early postnatal period. Often, the nurse is the first health care provider to examine a newborn after birth and must be aware of the signs of this condition to minimize the risk of testicular loss. PMID- 11012124 TI - Is motherhood good for women? A feminist exploration. AB - Motherhood as biologic destiny for women, has been long assumed. As nurses who aid women struggling with the demands of career and motherhood, we need to question this assumption. An exploration of feminist theory offers nurses ideas (and perhaps permission) to question the purpose and effects of mothering and the effects of mothering on women in our culture. Additionally, parallels between motherhood and the nursing profession are drawn. PMID- 11012125 TI - An inpatient cervical cancer screening program to reach underserved women. AB - Underserved women (e.g., African American, older women of all races) are less likely to be screened than other groups. The Johns Hopkins Hospital began the Cervical Cancer Screening Program (CCSP) to provide direct Papanicolaou (Pap) test screening to inpatient females. Over a 1-year period, 256 women received Pap tests as part of the CCSP. Of these, 56% (n = 144) were African American, 36% (n = 92) received medical assistance benefits, and only 47% (n = 120) had health insurance. The CCSP is an effective way to screen low-income and underserved females who otherwise may not be screened. PMID- 11012126 TI - Prenatal perineal massage: preventing lacerations during delivery. AB - OBJECTIVE: To investigate the associations between perineal lacerations and 13 variables associated with the incidence of perineal lacerations. Of particular interest was the variable of prenatal preparation of the perineum. DESIGN: This retrospective descriptive study used a convenience sample of 368 women whose delivery was attended by at least one of two midwives practicing in the Northwest between 1979 and 1995. SETTING AND PARTICIPANTS: All births in the study occurred in a home-based midwifery practice in the Northwest. The sample was primarily white and included 307 multiparous and 61 primiparous women. MAIN OUTCOME MEASURES: The initial chi squares indicated that five of the 13 factors investigated were significantly associated with the degree of laceration: parity, maternal age, maternal position at delivery, length of second stage of labor, and prenatal perineal massage. However, further analyses showed that when parity was controlled, the only factors independently associated with the seriousness of lacerations were parity and prenatal perineal massage. CONCLUSION: This study supports the conclusion that teaching perineal massage to primiparous women and multiparae who had episiotomies with their previous births is a useful intervention. It suggests that further study may help clarify the optimum frequency, timing, and technique of massage. PMID- 11012127 TI - Experienced obstetric nurses' decision-making in fetal risk situations. AB - OBJECTIVE: To determine which clinical factors experienced obstetric nurses consider most important in determining fetal risk during the intrapartum period. DESIGN: Ten dichotomized variables relevant to participants' clinical decision making were manipulated in fractional factorial vignettes. Participants were asked to rate the severity of fetal risk on a Likert scale after reading the vignettes. SETTING: About 87% of the participants worked in institutions with 4,000 or fewer deliveries per year. More than 25% worked in tertiary level facilities, and the remainder were employed in primary or secondary level facilities. PARTICIPANTS: Participants (N = 573), randomly selected from a list of nurses certified in "inpatient obstetric nursing" by the National Certification Corporation, were mailed the vignettes. The average number of years of intrapartum nursing experience was 13. MAIN OUTCOME MEASURE: Multiple regression analysis was used to determine the weights given to the 10 clinical factors by participants. RESULTS: Fetal scalp pH, maternal parity, amniotic fluid color, and long-term variability of the fetal heart rate were the most important predictors of nurses' fetal risk assessments. CONCLUSIONS: Long-term variability was the most important cardiotocographic factor in nurses' fetal risk assessments and the only cardiotocographic factor of the best 4 predictors of fetal risk. This indicated that participants were realistic about the limitations of electronic fetal monitoring (EFM). PMID- 11012128 TI - Identifying labor support actions of intrapartum nurses. AB - OBJECTIVE: To identify specific nursing actions that best characterize labor support from the nurse's perspective. DESIGN: A descriptive survey design using a three-round Delphi technique was used to explore the views of intrapartum nurses in the United States related to labor support. PARTICIPANTS: Five hundred AWHONN members who identified themselves as intrapartum nurses were invited to participate in the survey. One hundred eighty-six nurses agreed to participate. Participants who submitted usable surveys in each round: round one, n = 166; round two, n = 115; and round three, n = 117. Eighty-seven nurses participated in all three rounds of the survey. RESULTS: The participants identified 55 specific nursing actions as supportive care. These interventions ranged from psychosocial support such as remaining with the mother if she is fearful to physical support measures such as position changes. The nurses clearly distinguished between supportive nursing care and the assessment and technical aspects of their job. The nurses also identified the overall goals of intrapartum nursing were to assure a safe outcome for the newborn (82.8% of participants) and for the mother (75% of participants). CONCLUSIONS: The supportive actions identified by this panel of intrapartum nurses were similar to ones identified by mothers in other studies. Nurses made a clear distinction between supportive nursing care and assessment skills or technical tasks. PMID- 11012129 TI - The circadian rhythm of blood pressure during pregnancy. AB - OBJECTIVE: To review the literature on the circadian rhythm of blood pressure during pregnancy. DATA SOURCES: Computerized searches on MEDLINE, CINAHL, and MIRLYN. STUDY SELECTION: Selected studies from 1 969 to 1997 were evaluated. DATA EXTRACTION: Data were extracted and information was organized under the following areas: definition of and the interconnection between circadian rhythm and blood pressure; the circadian variability of blood pressure throughout the trimesters; the patterns of the circadian rhythm of blood pressure in pregnancies defined as normal and those complicated by chronic hypertension and preeclampsia; and clinical implications. DATA SYNTHESIS: The circadian rhythm of blood pressure in pregnancy is the same as in the non-pregnant state, with a nocturnal decrease, especially during sleep. In patients with chronic hypertension, the nocturnal fall in blood pressure may be steeper. Patients with mild preeclampsia may experience a less pronounced nocturnal decrease in blood pressure. Patients with severe preeclampsia may display a reversed circadian rhythm, with no decrease and/or an increase in nocturnal blood pressure. CONCLUSIONS: The patterns of the circadian rhythm of blood pressure during normal pregnancy and pregnancies complicated by chronic hypertension and preeclampsia warrant consideration when monitoring patients and implementing management plans. PMID- 11012130 TI - The development of AWHONN's fetal heart monitoring principles and practices workshop. Association of Women's Health, Obstetric and Neonatal Nurses. AB - The Fetal Heart Monitoring Principles and Practices (FHMPP) workshop was designed at the request of AWHONN's Committee on Education to meet member requests for standardized education in fetal heart assessment, including didactic and psychomotor skill content. Events and decisions are chronicled, from inception of the FHMPP program to development, evaluation, and revision. Current statistics and participation demonstrate the success of the workshop and its ongoing need. Today, as the project continues to grow and improve, its primary strength lies in member involvement and support. PMID- 11012131 TI - Legal implications of fetal heart assessment. AB - The standard of care requires perinatal nurses to perform fetal heart (FH) assessment competently and safely. Failure to adhere to established guidelines and standards for FH assessment may result in negative outcomes for the fetus or newborn and contributes to claims of nursing negligence. The perinatal nurse must be fully cognizant of professional guidelines and standards for FH assessment and comply with agency policies and procedures when conducting assessment of the fetal heart. Guidelines for FH assessment during the antepartum and intrapartum period are discussed within the context of restructured health care settings and today's medicolegal climate. PMID- 11012132 TI - Computer analysis of the fetal heart rate. AB - Computer analysis of the fetal heart rate is a technology of the Information Age commercially available for research and clinical practice. Intelligent systems are engineered with algorithms or neural networks designed to simulate expert knowledge. Automated analysis has provided objective, standardized, and reproducible data used to research fetal heart rate responses in the antepartum and intrapartum setting. Perinatal information systems can integrate FHR analysis and data management. PMID- 11012133 TI - Fetal pulse oximetry: an adjunct to electronic fetal heart rate monitoring. AB - Electronic fetal heart rate monitoring is routinely used as an indirect measure of fetal oxygenation, yet its value continues to be questioned. With a nonreassuring fetal heart rate pattern, the clinician often needs additional information about fetal oxygen status. Fetal pulse oximetry is a new fetal assessment technology. After consideration of the results of a multicenter randomized, controlled, clinical trial on fetal pulse oximetry in the United States, the U.S. Food and Drug Administration approved the technology for clinical use on May 12, 2000. The results of this trial are anticipated to be published in late 2000. PMID- 11012134 TI - Hypoxic-ischemic encephalopathy. AB - Cerebral hypoxia-ischemia (asphyxia) occurring in the fetus and newborn infant is a major cause of acute mortality and chronic neurological disability in survivors. This review highlights many practical aspects of perinatal hypoxic ischemic brain damage, including neuropathological features, obstetrical antecedents, and clinically important aspects of identification, management, and prognosis. Diagnostic techniques, including neuro-imaging, to diagnose hypoxic ischemic encephalopathy also are discussed. A thorough knowledge of the clinical spectrum of perinatal hypoxic-ischemic encephalopathy should enable neonatologists to undertake appropriate management strategies and prognostic indicators. PMID- 11012135 TI - Congenital chylothorax. AB - The objective of this paper is a retrospective study of all infants treated for congenital chylothorax at the Royal Children's Hospital (RCH), Melbourne, Australia and King Fahad National Guard Hospital (KFNGH), Riyadh, Kingdom of Saudi Arabia. The charts of all infants with congenital chylothorax admitted to RCH over a period of 13 years, June 1982-August 1994, and admissions to KFNGH over a 7-year period, June 1992-August 1998 inclusive, were reviewed including management outcome and complications. There were 19 infants, 13 from RCH and 6 from KFNGH; 11 females and 8 males. Three infants were managed antenatally. Fifteen infants presented immediately after birth. Seven were born with hydrops fetalis, 6 infants had syndromes and 10 infants were born prematurely. Regular infant feeding formula and/or breast milk were used successfully in 12 infants, while in 7 infants medium chain triglycerides (MCT) rich formula was used. Sixteen infants were mechanically ventilated with 75% of them ventilated for < or = 28 days. Fifteen infants received total parenteral nutrition (TPN), and in 80% for < or = 32 days. Hydropic infants had longer duration of mechanical ventilation and hospital stay with mean (range) of 33.9 (3-120) and 115 (23-225) days, respectively, compared with 18 (1-62) and 34.3 (14-88) days for nonhydropic infants. Five infants underwent surgery with failure in four. Sepsis and bronchopulmonary dysplasia were the main complications. The survival rate was 100% regardless of the mode of therapy. The prognosis of Isolated congenital chylothorax in term, and preterm infants is good even in the presence of hydrops. Breast milk and/or regular infant feeding formula should be used initially before proceeding to MCT-rich formula, which may be necessary in some cases. Surgery should be considered if conservative management of congenital chylothorax fails after 4-5 weeks. PMID- 11012136 TI - Association between epidural analgesia and intrapartum fever. AB - The objective of this paper is to determine whether or not epidural analgesia is an independent risk factor for intrapartum fever. Maternal temperature was measured every 4 h during labor to 1004 consecutive women in term labor. Women with fever or on antibiotics were excluded. Epidural analgesia was administered upon patients' request. Of the 406 (40%) women who received epidural analgesia, 11.8% (n = 48) developed a fever > or = 37.8 degrees C during labor compared with only 0.2% (n = 1) of women not receiving epidural analgesia. Women who received epidural analgesia were more likely to have one or more risk factors for intrapartum infection. Their labor and ruptured membranes were longer, they were more likely to have internal monitoring and have more vaginal examinations. Compared with women who received epidural analgesia and did not develop intrapartum fever, women that did develop fever had longer epidurals and more risk factors for infection. However, in a logistic regression analysis with fever as dependent variable, only the duration of epidural was significantly associated with the occurrence of fever. The rate of fever increased with longer labors, from 5% with labor < 3 h to 28% with labor > 6 h. In 90% of women the fever resolved within a few hours after delivery. Sepsis evaluation was negative in all of the newborns to mother who had intrapartum fever. Our data support a noninfectious etiology for intrapartum fever in the vast majority of our patients. However, infection must be ruled out before a decision is made to withhold antibiotic therapy. PMID- 11012137 TI - Yield and costs of screening growth-retarded infants for torch infections. AB - Many infants with intrauterine growth retardation (IUGR) are screened for TORCH infections. The yield and costs of such a practice may not be justifiable. Medical charts of infants with IUGR who had a workup for toxoplasmosis, other (infections), rubella, cytomegalovirus (infection), and herpes (simplex) (titer) (TORCH) infections were reviewed for the presence of clinical findings, laboratory and head ultrasound abnormalities associated with intrauterine infections. Maternal charts and reports of placental pathology were reviewed for identifying maternal illnesses and placental causes associated with IUGR. Seventy five out of 182 infants (41%) with IUGR had a workup for TORCH infection. Maternal conditions associated with IUGR included: pregnancy-induced hypertension (19%), tobacco use (43%), alcohol abuse (21%), illicit drug use (24%), chronic hypertension, diabetic vasculopathy or collagen vascular disease (12%), and multiple gestation (3%). Placental pathology was available in 53/75 cases. Thirty six of fifty-three (67%) placentae had abnormalities associated with IUGR: placental infarcts (22 of 36), vasculitis/villitis (15 of 36), placenta previa (1 of 36), abruptio placenta (2 of 36), and velamentous insertion of umbilical cord (1 of 36). Clinical findings among infants included hepatosplenomegaly, cataract or rash (1 of 75), thrombocytopenia and/or neutropenia and/or direct hyperbilirubinemia (11 of 75). Seven out of 75 infants had dysmorphic features. None of the infants (0 of 75) had positive IgM titers for toxoplasma, rubella, cytomegalovirus (CMV), or herpes simplex virus (HSV). No infants (0 of 43) had elevated total IgM titers; one infant (1 of 57) had a positive urine culture for CMV. One infant had evidence of calcifications on head ultrasound and a second infant had hydrocephalus (2 of 43). The costs associated with workup for TORCH infections among 75 infants included: TORCH titers determination: $17,816, total IgM titers: $1318, urine culture for CMV: $5734, and head ultrasound: $28,165. The yield of workup for TORCH infection among infants with IUGR is poor and does not justify the incurred costs. PMID- 11012138 TI - Anemia during pregnancy and birth outcome: a meta-analysis. AB - To determine the relationship between maternal anemia [hemoglobin (Hgb) < 10-11 g/dL] and various birth outcomes, a meta-analysis was conducted based on published literature identified by MEDLINE and manual search from 1966 through 1999. Odds ratios (OR) from selected studies were pooled according to the gestational age at anemia diagnosis. The meta-analysis shows that maternal anemia during early pregnancy was associated with slightly increased preterm birth [pooled adjusted OR (aOR): 1.32, 95% confidence interval (CI): 1.01-1.74], and nonstatistically significant increased low birth weight [pooled aOR: 1.39 (0.70 2.74)], and was not associated with fetal growth restriction [pooled aOR: 1.01 (0.73-1.38)]. However, there was a nonstatistically significant inverse relationship between anemia during late pregnancy and preterm birth [pooled aOR: 0.92 (0.54-1.84)] and low birth weight [pooled aOR: 0.80 (0.64-1.00)]. Anemia was not statistically significantly associated with hypertensive disorders of pregnancy regardless of stage of pregnancy [pooled OR: 0.80 (0.53-1.20)]. The relationship between anemia and perinatal mortality was inconclusive. A few studies indicated that severe maternal anemia (Hgb < 8-8.5 g/dL) was associated with increased risk of poor outcomes. We conclude that early pregnancy anemia is associated with slightly increased risk of preterm birth. The trend toward an inverse association of anemia determined during late pregnancy with preterm birth and low birth weight may reflect the benefit of plasma volume expansion. PMID- 11012139 TI - Sandifer's syndrome in a breast-fed infant. AB - Sandifer's syndrome is a rare manifestation of gastroesophageal reflux (GER) in children, occurring in association with abnormal movements of the head, neck, and upper part of the trunk. Out of 65 children with Sandifer's syndrome described in literature, only 2 were breast-fed. We report on a 15-day-old breast-fed girl affected by Sandifer's syndrome. Pathological GER was diagnosed with 24 h pH esophageal monitoring. In our patient, all the symptoms of Sandifer's syndrome disappeared when she was cow's milk formula-fed. The role of food allergy to dietary proteins ingested by a lactating mother is discussed. PMID- 11012140 TI - Severe aortic thrombosis in the neonate--successful treatment with low-molecular weight heparin: two case reports and review of the literature. AB - Small- to moderate-sized aortic thrombi in the sick newborn are frequently demonstrated, however, severe aortic thrombosis is rarely encountered. We report two newborn infants presenting with signs of decreased lower limb perfusion shortly after birth. An occlusive infrarenal aortic thrombus was demonstrated in both infants by ultrasound. No predisposing condition for thrombosis could be found in the first infant; in the second, an umbilical arterial catheter was the likely cause. The infants were successfully treated with low-molecular-weight heparin. In both infants resolution of the thrombus was observed by ultrasound. No complications of treatment were encountered. The clinical spectrum of neonatal aortic thrombosis and treatment options are reviewed. PMID- 11012141 TI - Variations in neutrophil count in preterm infants with respiratory distress syndrome who subsequently developed chronic lung disease. AB - Neutrophil counts were studied in 62 preterm infants receiving mechanical ventilation for neonatal respiratory distress syndrome (NRDS). Exploratory analysis indicated that the severity of NRDS, as demonstrated by fractional inspired oxygen (FiO2), mean airway pressure (MAP), arterial-alveolar PO2 ratio (a/APO2) and oxygenation index (OI), was correlated with percentage change of neutrophil counts during the first 5 days of life. Further analysis demonstrated that infants with NRDS who subsequently developed chronic lung disease (CLD) (n = 21) had statistically significant differences in variation of neutrophil counts when compared with the remainder (n = 41) without CLD (-35.0% +/- 4.3 vs. -16.9% +/- 5.8, p < 0.02). It is concluded that significant variations in neutrophil counts during the first 5 days of life may be found in infants with NRDS who subsequently develop CLD and that these changes may have predictive value regarding the development of CLD. PMID- 11012142 TI - Mode of delivery and neonatal hematocrit. AB - The objective of this paper is to test the hypothesis that the 2-h hematocrit (HCT) is lower in infants born by cesarean section (CS) than in infants born vaginally and that the postnatal rise of HCT is lower in infants born by CS than in infants delivered vaginally. We prospectively studied 31 infants delivered by elective CS, 21 infants delivered by CS because of arrest of descent and 30 vaginally delivered (VD) infants. All pregnancies were term, uncomplicated in nonsmoking mothers. Apgar scores were > 7 at 1 and 5 min. In all infants umbilical cord was clamped early. Umbilical vein and 2-h peripheral venous micro HCT were measured by centrifugation. Cord-blood HCT, 2-h HCT, and rise in HCT were similar in the two CS groups and significantly lower than in the VD group. In multiple regression, gestational age, Apgar score, or the presence or not of labor did not influence cord-blood HCT, 2-h HCT, or the rise in HCT. Infants born by CS have lower HCT than infants born vaginally. Prediction of 2-h HCT from cord blood HCT must take into account the mode of delivery. PMID- 11012143 TI - A tribute to Theodor M. Fliedner. PMID- 11012144 TI - Regulatory mechanisms controlling hematopoiesis: principles and problems. AB - Hematopoiesis is regulated by the combined action of specialized stromal cells and a consortium of hematopoietic regulatory factors. The multiplicity of these regulatory controls does result in overlapping regulator action, but multiple regulators are required to stimulate stem cell proliferation and are more efficient than single regulators when stimulation of progenitor cells is required. Gene inactivation studies have indicated that despite overlapping actions each hematopoietic regulator does have unique functions. Delayed elevations of stem and progenitor cells in the blood are a feature of enhanced hematopoiesis induced by the injection of regulators. These cells are not a random sample of marrow cells in such situations and may well be selected to rapidly amplify hematopoiesis by seeding previously inactive hematopoietic regions. PMID- 11012145 TI - The role of blood stem cells in hematopoietic cell renewal. AB - It has been the purpose of this keynote address to review available evidence for the notion that the stem and progenitor cells circulating in the peripheral blood play a decisive role in the homeostasis of blood cell formation distributed throughout dozens of bone marrow units in the skeleton. Furthermore, if this notion is correct, one could speculate that the quantity and quality of stem and progenitor cells in the blood should reflect the functional state of the hematopoietic stem cell system throughout the skeletal bone marrow and provide a new tool for the evaluation of alteration in blood cell production. On this basis, the following questions are considered: A) What do we know about the quality and quantity of blood stem cells in steady state conditions? B) In what way do blood stem cells respond to perturbations of the "steady state" of blood cell formation? C) Which role do blood stem cells play during hemopoietic development assuming that the establishment of bone marrow hemopoiesis requires the "seeding" of blood stem cells into an appropriate cellular environment? D) What is the role of blood stem cells in hemopoietic regeneration after partial body irradiation with a small volume of marrow (and hence stem cells) protected? and E) What are the mechanisms and/or kinetics of hemopoietic recovery if stem cells introduced into the circulation were collected from exogenous (autologous or allogeneic) sources? In this review presentation, experimental work of our group and of other members of the scientific community is summarized. It becomes obvious that blood stem and progenitor cells play a key role in hematopoietic homeostasis. Furthermore, their physiology and pathophysiology deserve rigorous experimental studies in order to develop a novel tool in the diagnosis and prognosis of neoplastic and non-neoplastic disorders of blood cell formation. PMID- 11012146 TI - Phenotype of the engrafting stem cell in mice. AB - The present data on engraftment into non-myeloablated mice strongly suggest that engraftment is determined by host-donor ratios as opposed to opening space. Theoretically, if the ratios of donor to host stem cells could be altered, especially without causing toxicity to the host animal, then the phenotypic readout could be increased in a clinically applicable manner. To research this further, we investigated low-dose irradiation (100 cGy) for its effects on marrow, spleen and peripheral blood counts, as well as engrafting stem cell levels. We found a transient but significant depression in the white blood cell and platelet counts in the peripheral blood which returned to normal by two weeks, with no apparent deleterious effect on the animals. However, the same irradiation dose after two months impaired marrow repopulation and reduced engraftment potential to less than 20% capacity. These results suggested that we could obtain much higher phenotypic readouts after engraftment with this model; thus, we assessed the engraftment of 40 million male BALB/c marrow cells into female hosts exposed to 100 cGy at two, five and eight months after cell infusion. The resultant high levels of chimerism, reaching 100% in many cases, strongly suggest that the key to engraftment in these models is host-donor stem cell ratios. One important issue relative to the above finding is whether cytokine-stimulated proliferating stem cells have irreversibly lost engraftment capacity or whether changes in the engraftment capacity are of a plastic nature, possibly related to cell cycle transit. A number of experiments following engraftment have shown that the engraftment defect is reversible and can be repeatedly lost and regained during the initial portions of a cytokine-stimulated culture. The above results suggest that, at least at the more primitive stem cell level, hematopoietic stem cell regulation may in part be based on a cell cycle model rather than a hierarchical system. PMID- 11012147 TI - Comparative effects of retroviral-mediated gene transfer into primary human stromal cells of Flt3-ligand, interleukin 3 and GM-CSF on production of cord blood progenitor cells in long-term culture. AB - The effects of different cytokines on growth of human cord blood CD34 cells was studied by performing long-term culture (LTC) with primary human stromal cells transduced with genes for either Flt3-ligand (L) (human transmembrane, murine soluble or murine membrane-bound forms), human interleukin 3 (IL-3) or human GM CSF. Molecular analysis of genomic DNA from transduced stromal cells using neo specific polymerase chain reaction demonstrated gene transfer of G418-selected stromal cell populations. Enzyme-linked immunosorbent assay and biological assays of conditioned media from transduced stromal cells indicated expression and release of soluble cytokines. Numbers of both immature and more mature progenitors (colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte; CFU-GEMM, BFU-E, CFU-GM) were increased threefold compared to control in the Flt3-L (transmembrane) LTC throughout five weeks of culture. IL-3 and GM-CSF feeders increased progenitor cell output also, but these effects were significantly lower than Flt3-L feeders. The two Flt3-L isoform engineered feeders, Ex6 (soluble isoform) and 5H (membrane-bound isoform), showed a decreased effect compared to the transmembrane Flt3-L feeders and, in particular, Ex6 feeders were similar to control feeders and 5H feeders were comparable to Flt3-L feeders only in the first two weeks of LTC. These results were apparent also by limiting dilution assays that showed a higher frequency of pre-CFU in the transmembrane Flt3-L feeders compared to control and the other cytokine feeders. Exogenous addition of soluble growth factors to suspension cultures without feeder layers, while superior to stromal feeders for short-term expansion of early progenitors, were inferior to the long-term maintenance/output on stromal feeders. Pre-CFU analysis supported these data. These results may be of some significance to understanding the actions of Flt3-L on blood cell production. PMID- 11012148 TI - Unilineage hematopoietic differentiation in bulk and single cell culture. AB - The rarity of hematopoietic stem and progenitor cells (HSCs, HPCs) has hampered the analysis of cellular and molecular mechanisms underlying early hematopoiesis. Methodology for HPC purification has partially offset this limitation. A further hurdle has been represented by the heterogeneity of the analyzed HPC/precursor populations: recently, development of unilineage HPC differentiation cultures has provided homogeneous populations of hematopoietic cells, particularly in the early differentiation state, i.e., populations pertaining to a single lineage and a restricted stage of differentiation/maturation, but sufficiently large for cellular/molecular analysis. This report focuses on the development and characterization of the unilineage HPC differentiation culture systems. A section is devoted to selected cellular and molecular mechanisms underlying hematopoiesis, which have been investigated by the HPC unilineage culture approach. Finally, recent advances in the development of HPC unilineage cultures at single cell level are discussed. PMID- 11012149 TI - Differences between normal and CML stem cells: potential targets for clinical exploitation. AB - Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder in which there is a deregulated amplification of CML progenitors at intermediate stages of their differentiation along the myeloid, erythroid and megakaryocyte pathways. Such cell populations are routinely quantified using standard in vitro colony forming cell (CFC) assays. The excessive production of leukemic CFC that is seen in most CML patients at diagnosis may be explained at least in part by their increased proliferative activity. An anomalous cycling behavior in vivo has also been found to extend to more primitive CML progenitor populations detectable as long-term culture-initiating cells (LTC-IC). Although the molecular basis of these changes in CML progenitor regulation is not fully understood at the level of the primitive CFC compartment, a selective inability of CML progenitors to be inhibited by certain -C-C-type chemokines has been demonstrated. Failure of the CML stem cell compartment to expand in vivo at the same rate as later progenitor cell types may be explained by their unique additional possession of an intrinsically upregulated probability of differentiation. Such a mechanism would be consistent with the observed loss of LTC-IC activity by CML cells incubated in vitro under conditions that sustain or expand normal LTC-IC populations. Initial clinical studies undertaken at our center established the feasibility of exploiting the differential behavior of primitive normal and CML cells in vitro as a potential purging strategy for reducing the leukemic stem cell content of CML marrow autografts. The results of a larger, second trial now in progress on a group of unselected patients are encouraging. Future studies of nonobese diabetic/severe-combined immunodeficiency mice engrafted with CML cells should provide another useful preclinical model for evaluating treatments that may more effectively eradicate the neoplastic clone in vivo. PMID- 11012150 TI - Maturation hierarchy of leukemic stem cells. AB - Here we discuss the characteristics of various progenitor cell assays for human acute myeloid leukemia (AML) with emphasis on the repopulating properties of human AML progenitor cells, their potential relevance for pathophysiological studies, and treatment development in AML. PMID- 11012151 TI - Human hematopoietic progenitor in bone marrow and peripheral blood. AB - The ability to enhance the frequency of progenitors in the peripheral circulation has resulted in peripheral blood progenitor cell collections being the preferred procurement method for autologous transplants and an accepted alternative for allografts. In autografts, peripheral blood-derived progenitor cells appear to have an advantage over conventional marrow transplants in the form of earlier engraftment without compromising sustained marrow function. Studies were performed to evaluate the broad class of CD34+ progenitors and their subtypes in preparations of bone marrow and peripheral blood cells. As a result, peripheral blood appears to contain a larger quantity of more mature progenitors, which is the likely reason for a more rapid return of granulocytes and platelets. However, controveries continue to exist as to whether the measurements derived from these studies can be used to predict time to engraftment and subsequent reestablishment of a marrow reserve. Some of these issues have been addressed in pilot studies where allografts using G-CSF-mobilized peripheral blood progenitor cells were compared to conventionally harvested bone marrow and bone marrow obtained from G CSF-stimulated donors. The time to engraftment of neutrophils and platelets were shortened when G-CSF-mobilized peripheral blood or marrow progenitor cells were used compared to case-matched grafts of steady-state marrow. The significance of the suggested differences between steady-state bone marrow grafts and mobilized peripheral blood cells remains to be determined. However, preparations of cytokine-mobilized peripheral blood progenitor cell preparations are being used with increasing frequency to replace conventional marrow harvests. PMID- 11012152 TI - Hemopoietic progenitor cells in the blood as indicators of the functional status of the bone marrow after total-body and partial-body irradiation: experiences from studies in dogs. AB - The granulocyte-macrophage colony-forming cells (GM-CFC) were studied in the blood of dogs to evaluate their relationship to the bone marrow GM-CFC under normal conditions and their involvement in hemopoietic regeneration after different types of exposure to ionizing radiation. The GM-CFC could be defined as regular blood elements showing characteristic levels of their concentration in individual dogs in the range from 20 to 300 cells per ml. In relative terms, the GM-CFC numbers present in the whole blood of normal dogs were found to be on the order of 0.1% of the GM-CFC numbers present in the bone marrow. A small fraction of the GM-CFC population in the bone marrow, i.e., about 1%, can be mobilized into the peripheral blood within three h by intravenous injection of dextran sulfate (DS). These cells are characterized by a small size and a low S-phase fraction similar to the GM-CFC that are normally present in the blood. Total-body irradiation with single doses of 0.8 Gy and more caused a characteristic pattern of sequential changes in the blood GM-CFC concentration that were related to the recovery of the bone marrow GM-CFC population. The blood GM-CFC concentration showed an extreme depression within the first 15 days, a transient increase from day 17 to day 35 and remained at subnormal values for several weeks and months. The regeneration of the GM-CFC population in the bone marrow that could be mobilized into the blood by DS was similarly delayed as the recovery of the blood GM-CFC values. In dogs which were kept under continuous radiation exposure (0.019 Gy/day) causing permanent damage to the hemopoietic system, the GM-CFC numbers in the blood remained permanently depressed. Partial-body irradiation of dogs with a myeloablative dose (11.7 Gy) given to the anterior part of their body was followed by sequential changes in the blood GM-CFC concentration specific for this type of exposure. The pattern of changes was determined by direct radiation effects, the compensatory responses in the protected bone marrow and the regeneration events in the irradiated bone marrow. On the other hand, it could be shown that the repopulation and the restoration of the hemopoietic tissue is initiated by the seeding of hemopoietic cells (including GM-CFC) from the protected marrow. PMID- 11012153 TI - Role of the CD34+ 38- cells in posttransplant hematopoietic recovery. AB - Using three different statistical tests in parallel, we showed in a preliminary study that neither mononuclear cells, CD34+ 33+ or 33- cells, nor CD34+ 38+ cells significantly correlated with engraftment kinetics following autologous blood cell transplantation (ABCT). We additionally demonstrated here, in a series of patients suffering from malignant diseases, that the graft content in CD34+ 38- cells is individually a more sensitive indicator of the earliest, as well as the latest post-ABCT trilineage hematopoietic recovery than the colony-forming units granulocyte-macrophage and even the total CD34+ cell content. This suggests that the CD34+ 38- cell population is itself subdivided into two more subsets, one being already lineage-committed and responsible for short-term engraftment, the other containing only very primitive hematopoietic cells responsible for sustained engraftment. Strong arguments favor the probability that these subsets correspond to HLA-DR+ and DR cells, respectively. We also defined an optimal threshold value of 0.05 x 10(6) CD34+ 38- cells/kg of the patient's body weight (b.w.) above which a rapid and sustained trilineage engraftment safely occurs. In fact, infusion of lower numbers of cells seems to have a more significant impact on long-term compared to short-term neutrophil recovery and on platelet kinetics engraftment. We additionally looked for the eventual influence on engraftment time of the type of disease, and of post-ABCT administration of hematopoietic growth factors (HGF). When the type of disease appeared to have no influence on the engraftment time, posttransplant HGF administration significantly reduced the time to trilineage engraftment in patients transplanted with < 0.05 x 10(6) CD34+ 38- cells, thus justifying it in case of reinfusion of low numbers of CD34+ 38- cells. On the other hand, the administration of HGF after infusion of more than 0.05 x 10(6) CD34+ 38- cells/kg b.w. did not hasten more, or only very little, the engraftment time, thus becoming not only unprofitable for the patients but costly as well. PMID- 11012154 TI - Hematopoietic progenitor cells in the blood and bone marrow in various hematologic disorders. AB - Hematopoietic progenitor cells are present in the blood and the bone marrow. Changes in the numbers of hematopoietic progenitor cells reflect alteration of pluripotent stem cells. We discuss such changes in common hematologic diseases including aplastic anemia, paroxysmal nocturnal hemoglobinuria (PNH) and thalassemia. In aplastic anemia, the numbers of burst forming units-erythroid (BFU-E) and colony-forming units-granulocyte-macrophage (CFU-GM) are much decreased; the decrease still exists after recovery from therapy. In PNH, the numbers of progenitor cells are low, even in the presence of marrow hypercellularity. In thalassemia, the numbers of progenitor cells are much increased; more pronounced in splenectomized patients. PMID- 11012155 TI - In vivo expansion of the circulating stem cell pool. AB - Stem cell trafficking between extravascular marrow sites and circulating blood is an essential part of the blood stem cell transplantation technology. Recombinant human G-CSF (rHuG-CSF) is widely used for stem cell peripheralization alone or together with chemopriming mobilizing early and pluripotent CD34+ cell subsets. New cytokine/chemokine mobilization regimens are under investigation such as combined rHuG-CSF and rHu thrombopoietin, rHuG-CSF and interleukin 3, rHuG-CSF and rHu stem cell factor, rHuG-CSF and Flt-3 ligand, human macrophage inflammatory protein, interleukin 1, and interleukin 8. Modifying the adherence of CD34+ cells to extracellular matrix molecules is a new mechanism by which hematopoietic progenitor cells are released into the circulating blood. Blocking the alpha4beta1 integrin receptor on CD34+ progenitor cells by using monoclonal antibodies specific for the heterodimeric complex alpha4beta1 has been shown to further increase the circulating stem cell concentration when given following rHuG-CSF priming. The current clinical research is primarily focused on improving stem cell mobilization efficiency in heavily pretreated and poorly mobilizing patients, and to decrease adverse effects of cytokine treatment. PMID- 11012156 TI - Mobilization of blood stem cells. AB - Deliberately increasing the number of hematopoietic stem and progenitor cells in the circulation allows faster and more efficient collection of sufficient cells for transplantation in both the allogeneic and autologous settings. These mobilized stem cells, when transplanted, provide quicker hematopoietic recovery for the patient than do nonmobilized blood stem cells or steady-state marrow derived stem cells. Currently used clinical procedures to produce stem cell mobilization include administration of G-CSF or GM-CSF, either as single agents or in combination with myelosuppressive chemotherapy. Some autologous blood stem cell donors exhibit indifference to currently applied mobilization therapies. This failure to mobilize has been associated with prior stem cell toxic therapy, e.g., radiation therapy and chemotherapy, but the association is incomplete. The observation that occasional normal donors have failed to respond to mobilization therapy indicates that factors other than stem cell damage could also be involved. Recently, a murine model has provided evidence that a circulating factor inhibits mobilization in some settings. Preliminary investigations have suggested that a circulating factor may inhibit mobilization of human hematopoietic progenitor cells in some instances. Studies to identify this factor(s) are underway. The mechanisms of blood stem cell mobilization are still poorly understood and there continues to be the potential to improve this process. PMID- 11012157 TI - Mobilization and transplantation of peripheral blood stem cells. AB - Two hundred nineteen patients underwent peripheral blood stem cell (PBSC) transplantation from 1990 to 1997. Stem cells were mobilized with cyclophosphamide (CY), or with CY plus Taxol or etoposide, followed by cytokines, and collected when leukocyte counts > or = 1,000/microl, or when CD34+ counts > or = 20/microl. On average, four to five collections were needed to obtain sufficient PBSC for engraftment. When CD34+ counts were used, the average number of collections decreased from 5.4 to 4.2. A discrepancy was noted in the extraction ratios and number of collections that depended on the optical density (I/O) setting of the leukapheresis machine. Patients collected at a setting of 100 had higher extraction ratios and required fewer collections (mean = 2.7) than those collected at 150 (mean = 4.4). This result was unexpected, because the entire mononuclear cell layer is collected at the higher I/O setting. Further analysis revealed that a larger volume of red cells was collected at 150 than at 100. These procedures used a small-volume collection chamber, so the chamber was apparently overloaded by RBC at the higher setting. More rapid recovery of neutrophil counts and platelet counts was seen in PBSC transplants than in autologous marrow transplants; moreover, PBSC transplant patients required fewer RBC and platelet transfusions. Sixteen out of 21 normal donors for allogeneic PBSC transplants gave adequate collections (> 2.5 x 10(6) CD34+ cells/kg), but three donors failed to yield > or = 1.5 x 10(6) CD34 cells/kg. This suggests an inherent difference among certain normal donors that may make PBSC mobilization difficult. PMID- 11012158 TI - The role of endothelium in the regulation of hematopoietic stem cell migration. AB - Mobilization of hematopoietic progenitor cells appears to be a multifactorial process which is at least partially regulated at the level of bone marrow microvascular endothelium (BMEC). In order to study the regulation of progenitor cell migration by endothelium in vitro, methods have been developed to isolate BMEC from bone marrow aspirates. In addition, immortalized BMEC cell lines have been generated. Using an in vitro model of migration across bone marrow endothelium, we demonstrate that only a small number of more mature, committed progenitors migrate spontaneously. In this model, adhesion molecules of the beta2 integrin family and the corresponding endothelial ligands are involved. The low spontaneous migratory capacity suggests that, in addition to adhesion molecules which mediate direct cellular contacts, paracrine cytokines and chemokines may play a role in progenitor migration across endothelium. Growth-factor-stimulated hematopoietic cells can produce cytokines which act on endothelial cells (e.g., vascular endothelial growth factor, VEGF), modifying their motility, growth, permeability, and fenestration. Therefore, VEGF might be involved in the mobilization and homing of hematopoietic progenitor cells. Furthermore, transendothelial migration of progenitors in vitro is substantially enhanced by the chemokine stromal-cell-derived factor-1 (SDF-1), which is produced by bone marrow stromal cells. More primitive progenitors, which do not migrate spontaneously, also respond to this chemokine. We conclude that transendothelial progenitor cell migration is regulated by adhesion molecules, paracrine cytokines, and chemokines. Mobilizing hematopoietic growth factors stimulate proliferation of hematopoietic cells, which may indirectly result in changes of the local cytokine and chemokine milieu, adhesion molecule expression, and eventually the mobilization of hematopoietic progenitor cells. PMID- 11012159 TI - Rapidly mobilizable stem cells. Do they belong to a special subpopulation? AB - Some characteristics of rapidly mobilized stem cells as a possible distinct subset of the murine bone marrow stem cell population are overviewed. Some of the agents that rapidly mobilize stem cells are toxic and possibly act through disrupting anchorage to the microenvironment. The mobilization occurring days after cytostatics and/or colony-stimulating factors (CSF), however, is a consequence of increased production or differentiation. While stem cells (colony forming units-spleen; CFU-S) circulating normally in blood have low self-renewal capacity (SRC), SRC of rapidly mobilized CFU-S is closer to that of bone marrow stem cells and is similar to that of the late mobilized stem cells. The survival rate of mice after transplantation of rapidly mobilized stem cells did not differ from that of bone marrow stem cells. One year after transplantation of rapidly mobilized stem cells, the SRC value of bone marrow did not differ from those transplanted with bone marrow cells. Replacement of a rapidly mobilizable stem cell pool requires 48 h under physiological conditions and a longer time after damage to hemopoiesis (irradiation, hydroxyurea injection). Possible physiological mechanisms in the anchorage of stem cells are discussed. PMID- 11012160 TI - Changes in the cytokine regulation of stem cell self-renewal during ontogeny. AB - The last 10 years have seen the development of a quantitative assay that is specific for transplantable totipotent murine hematopoietic cells with durable in vivo blood-forming ability. Recently, this assay has been successfully adapted to allow the detection and enumeration of an analogous population of human hematopoietic stem cells using myelosuppressed immunodeficient (nonobese diabetic/severe-combined immunodeficiency) mice as recipients. Characterization of the cells detected by this assay indicates their close relationship in both mice and humans with cells detected in vitro as long-term culture-initiating cells (LTC-IC). Culture conditions have now been identified that support a significant net expansion of these cells from both species. More detailed analyses of the cytokine requirements for this response indicate that the viability, mitogenesis and maintenance of LTC-IC function by human CD34+ CD38- cells can be independently regulated by exogenous factors. Superimposed on this uncoupling of hematopoietic stem cell "self-renewal" and proliferation control is a change during ontogeny in the particular cytokines that regulate their responses. These findings unite stochastic and deterministic models of hematopoietic stem cell control through the concept of a molecular mechanism that actively blocks stem cell differentiation and must be maintained when these cells are stimulated to divide by exposure to certain types and concentrations of cytokines. PMID- 11012161 TI - In vivo expansion of hemopoietic stem cells. AB - Under conditions of steady-state hemopoiesis, a small fraction of immature hemopoietic cells, including stem cells, circulates in peripheral blood (PB). In rhesus monkeys, a median number of 1.2 x 10(7)/l CD34+ cells was observed as opposed to a median number of 1.5 x 10(9)/l in aspirated bone marrow (BM). The concentration of circulating CD34+ cells is therefore approximately two logs less than that in BM. Since a 4-kg rhesus monkey has an estimated number of 3 x 10(10) BM cells and approximately 300 ml of blood, the fraction of CD34+ cells that circulates can be estimated at approximately 0.4% of the total pool of CD34+ cells. During hemopoietic reconstitution following a cytotoxic insult such as results from a midlethal dose of TBI, PB CD34+ cell numbers appeared to be correlated to those of BM, suggesting that PB CD34+ cells may reflect reconstitution of BM CD34+ cells. Reconstitution of BM immature cells can be accelerated by treatment with pharmacological doses of growth factors, resulting in largely expanded immature cell populations within a few weeks after TBI. Growth factors observed to exert such an effect included, notably, thrombopoietin. Such an acceleration can be monitored by daily assessment of circulating CD34+ cells. Expansion of immature circulating cells indicates expansion of similar cells in the bone marrow rather than growth factor-induced selective mobilization of immature cells. PMID- 11012162 TI - Development of natural killer cells from lymphohematopoietic progenitors of murine fetal liver. AB - We established a clonal culture system which supports the growth of murine immature natural killer (NK) cells. When we plated day 14 fetal thymocytes in methylcellulose media containing interleukin 2 (IL-2), IL-7 and steel factor (SF), we observed diffuse colonies which could not be classified into known colony types. Cells in the colonies were blast-like and expressed Thy-1 and CD25 but not lineage-specific markers. Cells in the colonies developed into NK1.1+ cells in fetal thymus organ culture indicating that the colonies consist of immature NK cells. We then examined the colony-forming ability of fetal liver cells. The combination of IL-2, IL-7 and SF with or without IL-11 supported formation of few immature NK cell colonies from purified progenitors. Interestingly, addition of IL-11 to the culture stimulated formation of mixed colonies consisting of immature NK cells, B cells, macrophages and/or mast cells. The clonal origin of the mixed NK cell colonies was confirmed by micromanipulation of the colony-forming cells. This culture assay should facilitate the analysis of the pathway and cytokine regulation of NK cell development. PMID- 11012163 TI - Ex vivo manipulation of hematopoietic stem and progenitor cells. AB - Approaches to manipulate peripheral blood progenitor cells (PBPC) ex vivo currently include the selection of CD34+ cells as a means to purge contaminating tumor cells from leukapheresis preparations or to provide a homogeneous starting population for the expansion of hematopoietic progenitor cells as well as the induction of postprogenitor cells of either the myeloid or megakaryocytic lineage. The latter cell populations might be used for an additional transplantation together with PBPC to possibly shorten the period of aplasia. In addition, ex vivo expansion of CD34+ cells can be used to generate autologous tumor-antigen-presenting dendritic cells for immunotherapeutic approaches aiming to treat minimal residual disease following high-dose chemotherapy. PMID- 11012164 TI - T lymphocytes determine the development of xeno GVHD and of human hemopoiesis in NOD/SCID mice following human umbilical cord blood transplantation. AB - Over the past decade the human-immunodeficient mouse chimera has become a well established in vivo model for studying the human immune system and/or hemopoiesis. Under certain experimental conditions and depending on the composition of the human cell graft, the recipient mice may develop a fatal disease, designated as discordant xenogenic graft-versus-host-disease (GVHD), which differs in target tissues and histopathology from allogenic GVHD. Experimental evidence is presented that immunodeficient mice are equally susceptible to allogenic GVHD as normal immunocompetent mice. Whole human cord blood and distinct cellular subpopulations from a single cord blood harvest were transplanted in NOD/severe combined immunodeficient mice and the repopulation of human cells was monitored over time. Depending on the ratio of lymphocytes to hemopoietic stem cells, proliferation of human T cells, hemopoiesis or a combination of the two is observed in widely varying proportions. When the graft contains a preponderance of lymphocytes, fatal protracted discordant xenogenic GVHD develops. Mice receiving purified CD34 cells survived up to 207 days in good health with more than 95% human cells in the bone marrow. In those mice all lineages (B and T lymphocytes, monocytes, granulocytes, erythrocytes and thrombocytes) were demonstrated in the bone marrow and peripheral blood. PMID- 11012165 TI - Amplification of true hemopoietic stem cells: possibilities and limitations. PMID- 11012167 TI - New vectors for gene therapy. AB - Retrovirus-based vectors are presently the most efficient gene transfer vehicles for introducing genes into human hematopoietic stem and progenitor cells. However, their use for gene therapy is still problematic. A major obstacle is viral sequences such as the tRNA primer binding site or the dimerization and encapsidation signals that are not required for the expression of the therapeutic gene. These sequences can recombine with endogenous and/or exogenous retroviruses to generate new forms of unpredictable retroviruses. Moreover, these sequences are the targets for transcriptional repressors which inhibit the expression of the transduced genes. Therefore we have developed a new generation of retrovirus vectors which self-delete upon integration. The vectors are based on the natural life cycle of retroviruses, involving duplication of the terminal control regions U5 and U3 to generate long terminal repeats, and on the ability of the P1-phage site-specific recombinase (Cre) to excise any sequences positioned between two target sequences (loxP). Thus, while inserting a therapy gene into the genome, the vectors simultaneously excise most proviral sequences that are not required for gene expression. PMID- 11012166 TI - Retroviral vectors containing a variant dihydrofolate reductase gene for drug protection and in vivo selection of hematopoietic cells. AB - Transfer of drug resistance genes to hematopoietic cells is being studied as a means to protect against the myelosuppression associated with cancer chemotherapy and as a strategy for the in vivo selection and amplification of genetically modified cells. The goal of this study was to test if retroviral-mediated gene transfer of a dihydrofolate reductase (DHFR) variant (L22Y) could be used for in vivo selection of transduced myeloid cells and to determine what proportion of transduced cells was required for protection from myelosuppression. Based on previous work suggesting that selection with antifolates may also require inhibition of nucleoside transport mechanisms, mice transplanted with DHFR transduced bone marrow cells were treated with trimetrexate and the nucleoside transport inhibitor prodrug nitrobenzylmercaptopurine riboside phosphate. In vivo selection of transduced myeloid progenitors was seen in the bone marrow and in circulating mature peripheral blood cells following drug treatment. These results show that the novel combination of the L22Y-DHFR cDNA, trimetrexate and nitrobenzylmercaptopurine riboside phosphate can be used to select for transduced myeloid cells, and that this approach warrants further study in large animal models. A bicistronic vector containing a human CD24 reporter gene was used to determine the number of modified cells needed for chemoprotection. Partial protection from neutropenia was seen when greater than 10% of myeloid cells expressed the vector, and high levels of protection were obtained when the proportion exceeded 30%. These results suggest that gene transfer may be useful for myeloprotection in certain pediatric cancers, but that more efficient gene transfer will be required to apply this approach to adult cancer patients. PMID- 11012168 TI - Results of retroviral and adenoviral approaches to cancer gene therapy. AB - Genetic modification for cancer treatment has involved the introduction of chemotherapy protection and sensitization genes into normal and tumor cells, respectively, for the purpose of improving the outcome of conventional approaches to the treatment of solid tumor neoplasms. This paper will review the use of multidrug resistance-1 retroviral vectors and cytosine deaminase adenoviral prodrug activation vectors for this purpose. PMID- 11012169 TI - Cytokine gene transfer in cancer therapy. AB - New strategies based on gene transfer technology are employed in cancer therapy. Cytokines are polypeptides involved in immunity and inflammation, and essentially control the magnitude of the immune response. Genetically modified tumor cells releasing various cytokines have been shown to enhance tumor immunogenicity and to induce the regression of preexisting tumors. In some instances, immunological memory has been generated to resist the subsequent challenge with unmodified, parental tumor cells. Cytokine gene transfer into antitumor effector cells, as well as antigen presenting cells, is also being investigated to augment antitumor immune responses. PMID- 11012171 TI - Trombopoietin: from molecule to medicine. Preface. PMID- 11012170 TI - Cover art: the watercolor drawings of James Homer Wright. PMID- 11012172 TI - The molecular control of hematopoiesis: progress and problems with gene manipulation. AB - The in vitro-based discovery and characterization of hematopoietic regulators were of great value in identifying many of the agents active in controlling hematopoiesis. Subsequent in vivo studies have validated most of the information obtained from the in vitro studies, although the in vitro studies proved to be somewhat misleading in predicting which agents would exhibit the greatest quantitative effects in vivo. Establishing more clearly the actual situation in vivo has required a return to more complex, and often less satisfactory, studies on genetically-manipulated whole animals. Of the two possible general approaches, gene inactivation models have proved more informative than transgenic, overexpression models. Each model has raised multiple questions in need of further resolution and the deletion studies have also indicated that other regulators must exist for various lineages, but have yet to be discovered. Of particular interest is the finding from gene inactivation studies that both G-CSF and thrombopoietin are necessary for the maintenance of normal numbers of progenitor cells in multiple lineages, suggesting that each of these lineage dominant regulators may have broader actions when operating on cells in the stem cell and progenitor cell compartments. PMID- 11012173 TI - Native thrombopoietin: structure and function. AB - Thrombopoietin (TPO), the c-Mpl ligand, is produced constitutively in liver and other organs, circulates in the bloodstream, and is delivered to bone marrow, where it stimulates the early development of multiple hematopoietic lineages and megakaryocytopoiesis. The concentration of TPO in blood is regulated by c-Mpl mass on platelets and megakaryocytes. In addition to regulation by the number of TPO molecules, including the possible modulation of TPO mRNA abundance in bone marrow, megakaryocytopoiesis and platelet production may be regulated as a result of modulation of TPO activity by proteolytic processing that generates truncated forms of the molecule. Characterization of TPO partially purified from human plasma, however, revealed that the full-length molecule was the predominant form in the blood of both normal individuals and thrombocytopenic patients, although small amounts of truncated species were detected. Thus, truncation of TPO, at least that in the circulation examined, does not appear to contribute to the direct regulation of platelet production in response to increased demand. Given that native TPO isolated from the plasma of thrombocytopenic animals comprises truncated forms, the truncation of TPO is likely of physiological importance in the life history of this molecule. PMID- 11012174 TI - Peptide agonists of the thrombopoietin receptor. AB - We have screened a variety of L-amino acid peptide libraries against the extracellular domain of the human thrombopoietin (HuTPO) receptor, c-Mpl. A large number of peptide ligands were recovered and categorized into two families. Peptides from each family compete with the binding of HuTPO and with the binding of peptides from the other familiy. Representative peptides were synthesized and found to activate the full-length HuTPO receptor expressed in Ba/F3 cells to promote proliferation. These peptide families show no apparent homology to the primary sequence of TPO. We have focused our optimization efforts on one of the peptides, a linear 14-mer (IEGPTLRQWLAARA) with an IC50 of 2 nM in a competition binding assay and an EC50 of 400 nM in the proliferation assay. In order to enhance the potency of the compound, we constructed dimeric peptides by linking the carboxy-termini of the 14-mers to a lysine branch. These molecules exhibited slightly higher affinity (0.5 nM) and greatly increased potency (0.1 nM). The EC50 of the dimeric peptide was equivalent to that of the 332 aa form of baculovirus-expressed recombinant HuTPO. As previously shown for the erythropoietin-mimetic peptides, the TPO-mimetic peptides probably activate the TPO receptor by binding and inducing receptor dimerization. This supposition is supported by the observation that covalent dimerization of the peptide enhances its potency by 4,000-fold over that of the monomer. The peptide dimer is also active in stimulating in vitro proliferation of progenitors and maturation of megakaryocytes from human bone marrow, and in promoting an increase in platelet count when administered to normal mice. PMID- 11012175 TI - IL-3 does not contribute to platelet production in c-Mpl-deficient mice. AB - Thrombopoietin is a lineage-dominant cytokine involved primarily in the control of platelet production. The physiological importance of thrombopoietin (TPO) in the regulation of megakaryocyte and platelet production was demonstrated by the production of mice deficient in TPO or its receptor, c-Mpl. Even though these mice are profoundly thrombocytopenic they maintain a basal level of approximately 10% of the normal count of fully functional platelets. These platelets prevent any abnormal bleeding episodes and highlight the potential importance of other factors in the control of platelet production. Among the factors with in vitro megakaryocytopoietic activity, the most potent is undoubtedly interleukin 3 (IL 3). To analyze the contribution of IL-3 to platelet formation in the absence of TPO, we have generated mice deficient in both c-Mpl and IL-3Ralpha by taking advantage of a natural mutation present in this gene in the A/J mouse. Surprisingly, these double knockout mice did not show any further reduction in their platelet or megakaryocyte counts when compared with c-Mpl-deficient mice. Similarly, progenitors from other lineages that are also reduced in c-Mpl deficient mice are not further affected by the absence of a functional IL-3Ralpha gene. These results demonstrate that IL-3 alone is not responsible for the production of a basal level of normal platelets in the absence of thrombopoietin signaling. PMID- 11012176 TI - Compared effects of Mpl ligand and other cytokines on human MK differentiation. AB - The discovery of the Mpl ligand (Mpl-L), also called thrombopoietin (TPO), has facilitated in vitro investigation of human megakaryocytopoiesis. By confocal microscopy, endomitosis appeared as abortive mitosis skipping late stages of mitosis. No telophase and cytokinesis were observed. A spherical multipolar spindle which limits chromatid segregation was observed. The nuclear envelope subsequently reformed isolating all chromatids in a single nucleus. Platelet shedding was ultrastructurally studied. Platelet release occurred after formation of long cytoplasmic extensions (proplatelet formation), constriction areas delineating platelet territories. Heterogeneity in platelet size may be determined by the length of these extensions. Pegylated-recombinant human megakaryocyte growth and development factor, a truncated form of Mpl-L, was the most efficient cytokine to produce proplatelet-bearing megakaryocytes (MKs) and platelets in vitro. However, functional platelets with a normal ultrastructure could be produced in the presence of a combination of other cytokines. Finally, we investigated whether the induction of MK differentiation by the MS-5 stromal cell lines is due to Mpl-L. MS-5 cells synthesized Mpl-L transcripts and a biologically active protein. When human CD34+ cells were grown in contact or noncontact cultures with MS-5 cells, MK differentiation was observed. Soluble Mpl receptor (sMpl-Fc) addition inhibited MK growth, suggesting that the MK-promoting activity was due to Mpl-L production. Marrow stromal cell lines derived from TPO /- mice were also able to sustain MK growth. Despite the absence of any production of Mpl-L, the sMpl-Fc continued to inhibit MK differentiation. This result suggests that the sMpl has a direct inhibitory effect and may explain the divergent results in the literature concerning the precise role of Mpl-L on the MK terminal differentiation. PMID- 11012177 TI - The role of IL-II in hematopoiesis as revealed by a targeted mutation of its receptor. AB - Interleukin 11 (IL-11) is a pleiotropic growth factor with several actions in common with members of the IL-6 family. IL-11 utilizes a specific receptor chain encoded by two genes, IL-11Ra, which is expressed in hematopoietic and other tissues and, IL-11Ra2, which has a restricted pattern of expression. The actions of IL-11 in the hematopoietic compartment include support of multilineage and committed progenitors contributing to myeloid, erythroid, megakaryocyte, and lymphoid lineages. IL-11 demonstrates a prominent thrombopoietic activity which is being evaluated in clinical trials. In contrast to the multiple in vitro and in vivo effects of IL-11, mice with a targeted mutation of the IL-11Ra gene (IL 11Ra-/-) did not exhibit an overt hematological phenotype. Generation of a null phenotype was confirmed by independent assays. The numbers of progenitor cells of various lineages as well as their terminally differentiated progeny were undisturbed in the IL-11Ra-/- mice. In addition, the mutant mice were able to respond appropriately to increased demand in situations of hematopoietic stress. This study has highlighted the growth factor redundancy operative in the hematopoietic compartment, and in addition, has served to identify a critical action of IL-11 in nonhematopoietic organs. PMID- 11012178 TI - Use of the zebrafish (Danio rerio) to define hematopoiesis. AB - Hematopoiesis in the vertebrate is characterized by the induction of ventral mesoderm to form hematopoietic stem cells and the eventual differentiation of these progenitors to form the peripheral blood lineages. Several genes have been implicated in the differentiation and development of hematopoietic and vascular progenitor cells, yet our understanding of the discrete steps involved in the induction of these cells from the ventral mesoderm is still incomplete. One method of delineating these processes is based on the use of lower vertebrates. The zebrafish (Danio rerio) is an especially robust vertebrate system for both isolating and characterizing genes involved in these processes. Hematopoietic mutants have been generated with defects in many of the steps of both the primitive and definitive hematopoietic programs. Cloning of the genes that underlie these mutations should yield valuable details of hematopoiesis and may have therapeutic implications for bone marrow transplantation and stem cell gene therapy. PMID- 11012179 TI - Transcription factor GATA-1 in megakaryocyte development. AB - The transcription factor GATA-1 is specifically expressed in hematopoietic lineages. Prior gene knockout experiments established an essential role for GATA 1 in red blood cell production, but could not provide direct evidence with respect to a requirement in megakaryopoiesis. We summarize here recent lineage selective gene targeting in mice that establishes critical functions for GATA-1 in controlling megakaryocyte growth and maturation, and platelet production. GATA 1 megakaryocytes are delayed in their cellular maturation, exhibit marked hyperproliferation and generate fewer than normal, yet enlarged, platelets in vivo. Thus GATA-1 is a central regulator in both the erythroid and megakaryocytic lineages. PMID- 11012180 TI - Analysis of hematopoietic stem cell reprogramming with toxigenicity. AB - The molecular mechanisms by which a stem cell is committed to individual lineage are largely unknown. Two different models, though not mutually exclusive, are currently debated. The first describes the temporal and hierarchical coordination of lineage-specific transcriptional programs. The second suggests that multilineage genes are expressed in a self-renewing and undifferentiated cell prior to lineage commitment. To challenge these two models in in vivo-appropriate conditions, the expression of an exogenous toxigene was used to create transgenic animals in which an inducible, reversible cell knock-out at a specific stage of differentiation could be achieved. Both additional transgenesis using the megakaryocyte specific alphaIIb promoter and targeted transgenesis were used to express the herpes virus thymidine kinase (tk) gene in the megakaryocytic lineage. When the tk gene was targeted to the locus of the megakaryocyte-specific alphaIIb gene, a typical Glanzman thrombasthenic syndrome was created. Despite this bleeding disorder, the lack of expression of the alphaIIb gene did not affect the development of the mice. In both transgenic and targeted animals, all progenitor cells were sensitive to the effect of the gancyclovir (GCV), both in vivo and ex vivo. Long-term bone marrow cell cultures on stromal layers indicated that most of the very early progenitor cells expressed the enzyme. All the results obtained with this inducible toxic phenotype indicated that genetic programs that are in control of the expression of lineage-specific genes are operative in a totipotent stem cell prior to lineage commitment and strongly support the concept that stem cells express a multilineage transcriptome. PMID- 11012181 TI - Cellular and molecular biology of megakaryocyte differentiation in the absence of lineage-restricted transcription factors. AB - Targeted gene disruption of two distinct lineage-restricted hematopoietic transcription factors has provided useful insights into the transcriptional control of platelet production. Absence of either the basic leucine-zipper protein NF-E2 or of the zinc-finger protein GATA-1 in vivo results in severe thrombocytopenia secondary to distinct patterns of arrested megakaryocyte cytoplasmic maturation; in addition, megakaryocyte-selective loss of GATA-1 expression leads to dysregulated proliferation of progenitor cells. The ultrastructure of the defective megakaryocytes suggests that absence of the respective transcription factors impairs biogenesis of platelet-specific granules and proper development and organization of demarcation membranes. In particular, transcriptional targets of NF-E2 may be implicated in the very final stages of megakaryocyte differentiation, which involve the organization and release of platelets. Preliminary characterization of genes that are downregulated in NF-E2 /- megakaryocytes is in progress and is likely to lead to mechanistic insights into thrombocytopoiesis. PMID- 11012182 TI - Cyclin D3 and megakaryocyte development: exploration of a transgenic phenotype. AB - The roles of cell cycle regulatory proteins in megakaryocyte development are poorly understood. We have previously demonstrated that cyclin D3 is expressed in megakaryocytes and is induced upon treatment with Mpl ligand. Transgenic mice in which cyclin D3 is overexpressed in the megakaryocytic lineage show features similar to in vivo Mpl ligand treatment, including increased megakaryocyte number and ploidy. Terminal maturation and platelet production are not enhanced, however, and transgenic megakaryocytes show a defect in demarcation membrane development. We have examined expression of the transcription factor nuclear factor (NF)-E2, known to be involved in cytoplasmic maturation and platelet fragmentation, in these transgenic mice and controls treated with Mpl ligand. Our findings demonstrate marked induction of NF-E2 mRNA in control megakaryocytes in response to Mpl ligand, but no NF-E2 increase in transgenic cells, potentially explaining the lack of platelet increase in these transgenic mice. Transgenic megakaryocytes treated with Mpl ligand display a limited increase in NF-E2. In response to literature reports of Mpl ligand-induced transient increases in p21Cip1/WAF1 mRNA in polyploidizing megakaryocytic cell lines, we have examined p21 transcript levels in both normal and transgenic megakaryocytes. In normal mouse spleen, only a small percentage of megakaryocytes express detectable levels of p21 mRNA, with the majority of these cells expressing at high intensity. p21 levels are not affected by treatment with Mpl ligand, while the frequency of expressing cells increases transiently. Transgenic megakaryocytes exposed to Mpl ligand also show an increased frequency of p21-positive cells, and stimulation with Mpl ligand resulted in a further increase in this frequency. The nature of this effect will require further investigation. PMID- 11012184 TI - The effect of MGDF on platelet function and thrombosis in animal models. AB - Recombinant human megakaryocyte growth and development factor (rHuMGDF) is a recombinant form of the endogenous c-mpl ligand, thrombopoietin (TPO). rHuMGDF (and c-mpl ligands in general) can produce a measurable sensitization of platelets to known platelet agonists. Our laboratory has observed this sensitization in vitro in both platelet-rich plasma and whole blood and ex vivo in platelets from animals receiving rHuMGDF. Concurrently, clear increases in the tyrosine phosphorylation of Jak2 and c-mpl receptor can be observed both in vitro and ex vivo. To assess the in vivo prothrombotic potential of rHuMGDF, a rabbit carotid artery model of cyclic flow reduction (CFR) was used. Intravenous administration of platelet-sensitizing dosages of rHuMGDF had no effect on the CFR pattern, whereas control experiments demonstrated that the CFR pattern can be modulated by both platelet sensitizing (epinephrine) and antithrombotic (aspirin and ketanserin) agents. We conclude that thrombopoiesis can be observed at dosages that do not sensitize platelets, and further, that platelet-sensitizing dosages of rHuMGDF do not necessarily enhance platelet-dependent thrombosis in this model. PMID- 11012183 TI - Effects of Mpl ligands on platelet production and function in nonhuman primates. AB - Endogenous thrombopoietin (TPO) stimulates platelet production in nonhuman primates dose-dependentbyinducing megakaryocyte development from early marrow hematopoietic progenitors and subsequent proliferation and endoreduplication. Recombinant human TPO, nonpegylated or pegylated recombinant human megakaryocyte growth and development factor produce log-linear responses in peak peripheral platelet counts (or peripheral platelet mass turnover), platelet TPO receptor density, and marrow megakaryocyte volume, ploidy, number and mass. Mpl ligand therapy sustains normal peripheral platelet concentrations following myelosuppressive chemotherapy in baboons and corrects peripheral platelet counts in HIV-infected chimpanzees with severe thrombocytopenia. Whereas Mpl ligands do not directly induce platelet aggregation in vitro, they enhance aggregatory responsiveness of platelets to physiologic agonists both in vitro and transiently ex vivo following treatment with Mpl ligands. However, platelet recruitment into forming thrombus is not augmented by these agents when evaluated in quantitative rabbit or baboon models of platelet-dependent thrombus formation, except for the direct effect of platelet concentration per se. These findings indicate that appropriate dosing of these agents prevents thrombocytopenia without increasing the risk of platelet-dependent thrombo-occlusive complications. PMID- 11012185 TI - The efficacy of recombinant TPO in murine And nonhuman primate models for myelosuppression and stem cell transplantation. AB - Radiation-induced pancytopenia proved to be a suitable model system in mice and rhesus monkeys to study thrombopoietin (TPO) target cell range and efficacy. TPO was highly effective in rhesus monkeys exposed to the midlethal dose of 5-Gy (300 kV x-rays) TBI, a model in which it alleviated thrombocytopenia, promoted red cell reconstitution, accelerated reconstitution of immature CD34+ bone marrow (BM) cells and potentiated the response to growth factors such as GM-CSF and G CSF. The accelerated reconstitution of BM CD34+ cells appeared to be reflected by a similar rise in peripheral blood CD34+ cells, both being augmented by concomitant GM-CSF. However, TPO was ineffective following transplantation of limited numbers of autologous BM or highly purified stem cells in monkeys conditioned with 8-Gy TBI. In the 5-Gy model, a single dose of TPO 24 h after TBI was effective in preventing thrombocytopenia and was augmented by GM-CSF. The strong erythropoietic stimulation may result in iron depletion and TPO treatment should be accompanied by monitoring of iron status. In mice, similar observations were made and the importance of dose and dose schedule for stimulation of multilineage repopulating cells versus the lineage-dominant thrombopoietic response studied in detail. PMID- 11012186 TI - The combined administration of daniplestim and Mpl ligand augments the hematopoietic reconstitution observed with single cytokine administration in a nonhuman primate model of myelosuppression. AB - This study evaluated the ability of daniplestim, a high affinity interleukin 3 receptor agonist, to enhance the hematopoietic response of Mpl ligand (Mpl-L) administration in nonhuman primates following severe, radiation-induced myelosuppression. Rhesus monkeys were total body x-irradiated (TBI) to 600 cGy, midline tissue dose. Beginning on day 1 post-TBI, animals were s.c. administered daniplestim (100 microg/kg bid; n = 4), Mpl-L (10 microg/kg qd; n = 3), daniplestim (100 microg/kg bid) plus Mpl-L (10 microg/kg qd) (n = 4) or 0.1% autologous serum (AS) (n = 11) for 18 days. CBCs were monitored for 60 d after TBI. The duration of thrombocytopenia (platelet count; PLT <20,000/microl) was significantly decreased by the administration of daniplestim (6.5 d, p = .01), Mpl-L (3.0 d, p = .003) and the coadministered daniplestim/Mpl-L (1.3 d, p = .001) compared to controls (10.4 d). As monotherapy Mpl-L but not daniplestim significantly improved the PLT nadir (21,000/microl, p = .023 and 5,000/microl, p = .266, respectively) compared to the control (3,000/microl). The combined administration of daniplestim and Mpl-L significantly improved the PLT nadir (28,000/microl, p = .007) compared to both the control cohort (3,000/microl) and the daniplestim only cohort (5,000/microl, p = .043). Recovery of PLT to preirradiation values occurred earlier in the daniplestim only (d 21) or the daniplestim/Mpl-L cohorts (d 18) than in the Mpl-L only or control cohorts (d 28, d 29, respectively). The administration of daniplestim or Mpl-L alone neither shortened the duration of neutropenia (ANC<500/microl) compared to the controls (15.8 d, 16.0 d versus 16.2 d, respectively), nor improved the recovery time of neutrophils to baseline values (d 22, d 25, and d 23, respectively). The ANC nadir was significantly improved by daniplestim alone but not Mpl-L administration (76/microl, p = .001 and 50/microl, p = .093, respectively) compared to the controls (8/microl). Coadministration of daniplestim and Mpl-L significantly improved the ANC nadir (196/microl, p = .001) compared to either the AS- or the monotherapy-treated cohorts. Also the duration of neutropenia observed in the AS-controls (16.2 d) was significantly reduced in the daniplestim/Mpl-L cohort (10.8 d, p = .002). The combined administration of daniplestim and Mpl-L significantly improved hematopoietic recovery and further enhanced the stimulatory effect of cytokine monotherapy, as well as reducing clinical support requirements after radiation-induced bone marrow myelosuppression. PMID- 11012187 TI - Myelofibrosis: experimental models and human studies. AB - Thrombopoietin (TPO) is the central regulator of megakaryocytopoiesis and thrombocytopoiesis. Preclinical data and human studies have so far shown that the recombinant molecule is safe to administer and associated with very little toxicity. Nevertheless, different experimental animal models have revealed that a chronic exposure to very high doses of TPO could result in myeloproliferative syndromes with a spectrum of pathological features in common with human idiopathic myelofibrosis (PMF). A number of investigators have researched whether TPO or its receptor Mpl were involved in the pathogenesis of human myeloproliferative syndromes which are also characterized by a predominant megakaryocytic involvement, in PMF and primitive essential thrombocythemia. In both diseases, megakaryocyte (MK) progenitors develop autonomously in serum deprived cultures. This spontaneous MK development is also observed at limiting dilution demonstrating that MK escape the normal regulatory controls. Furthermore, this abnormal MK proliferation and maturation is neither due to an autocrine stimulation by TPO nor by point mutation or deletion in the coding region of the c-mpl gene. This paper will review the data that have been reported to date on the effects of an overexpression of Mpl ligand and related molecules on the induction of experimental myelofibrosis and highlight recent insights into the pathogenesis of PMF. PMID- 11012188 TI - Endogenous TPO (eTPO) levels in health and disease: possible clues for therapeutic intervention. AB - The factor which is the primary regulator of megakaryocyte and platelet production has recently been identified as the ligand for the receptor Mpl. This discovery has resulted in substantial advances in our understanding of platelet homeostasis. The access to new experimental reagents has enabled studies of the endogenous circulating form of this ligand, endogenous thrombopoietin, in normal individuals and in patients with altered platelet numbers. The relationship of endogenous TPO in health and disease will be examined with consideration of the implications for successful therapeutic intervention with exogenous recombinant Mpl ligands in selected settings. PMID- 11012189 TI - Defective c-Mpl signaling in the syndrome of thrombocytopenia with absent radii. AB - Thrombocytopenia with absent radii (TAR) syndrome is a rare congenital defect with severe hypomegakaryocytic thrombocytopenia and bilateral radial aplasia. To elucidate a possible relationship between thrombocytopenia in TAR and defects in the thrombopoietin (TPO)/c-Mpl system, we examined TPO activity in sera from six patients and in vitro reactivity of the patients' platelets to recombinant human TPO. We found elevated TPO serum levels in all patients, excluding a TPO production defect as a pathomechanism for the thrombocytopenia. In contrast to healthy controls, however, platelets of TAR patients failed to respond to recombinant TPO as measured by testing TPO synergism to suboptimal concentration of platelet activators. Most interestingly, TPO-induced tyrosine phosphorylation of platelet proteins was completely absent (four out of five) or markedly decreased (one out of five). More detailed investigations of the signal cascades of c-Mpl demonstrated the absence of Jak2 phosphorylation after TPO stimulation in a TAR patient's platelets. A defect in the early events of c-Mpl signal transduction might be the reason for impaired megakaryocytopoiesis in TAR syndrome. PMID- 11012190 TI - A novel thrombopoietin signaling defect in polycythemia vera platelets. AB - The pathogenesis of polycythemia vera (PV), a disease involving a multipotent hematopoietic progenitor cell, is unknown. Thrombopoietin (TPO) is a newly characterized hematopoietic growth factor which regulates the production of multipotent hematopoietic progenitor cells as well as platelets. To evaluate the possibility that an abnormality in TPO-mediated signal transduction might be involved in the pathogenesis of PV, we examined TPO-induced protein tyrosine phosphorylation using platelets as a surrogate model system. Platelets were isolated from the blood of patients with PV as well as from patients with other chronic myeloproliferative disorders and control subjects. Impaired TPO-mediated platelet protein tyrosine phosphorylation was a consistent observation in patients with PV as well as those with idiopathic myelofibrosis (IMF), in contrast to patients with essential thrombocytosis, chronic myelogenous leukemia, secondary erythrocytosis, iron deficiency anemia, hemochromatosis, or normal volunteers. Thrombin-mediated platelet protein tyrosine phosphorylation was intact in PV platelets as was expression of the appropriate tyrosine kinases and their cognate substrates. However, expression of the platelet TPO receptor, Mpl, as determined by immunoblotting, chemical crosslinking or flow cytometry was markedly reduced or absent in 34 of 34 PV patients and also in 13 of 14 IMF patients. Impaired TPO-induced protein tyrosine phosphorylation in PV and IMF platelets was uniformly associated with markedly reduced or absent expression of Mpl. We conclude that reduced expression of Mpl is a phenotypic characteristic of platelets from patients with PV and IMF. The abnormality appears to distinguish PV from other forms of erythrocytosis and may be involved in the platelet function defect associated with PV. PMID- 11012191 TI - Overview of the safety and biologic effects of PEG-rHuMGDF in clinical trials. AB - Completed randomized placebo-controlled phase I/II studies of pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) have demonstrated that this recombinant Mpl ligand has potent and lineage-dominant effects on megakaryopoiesis and platelet production. Platelets produced after PEG rHuMGDF administration display normal ultrastructure and functional attributes. In these early studies, PEG-rHuMGDF accelerated the recovery of baseline platelet counts after cytotoxic chemotherapy in cancer patients by six to seven days, indicating the potential for clinical benefit in this setting. PEG-rHuMGDF has been well-tolerated in clinical trials, with similar adverse events in placebo and PEG-rHuMGDF populations, and an observed adverse event profile consistent with the effects of underlying malignancy and chemotherapy. The lack of inflammatory cytokine effects in the clinic is consistent with results of animal studies, the narrow tissue distribution of Mpl and the lineage-dominant effect of PEG-rHuMGDF on megakaryopoiesis. Additional phase I/II studies have commenced in the fields of cancer chemotherapy and augmentation of platelet donation, and a phase III study is underway in patients undergoing bone marrow transplantation. PMID- 11012192 TI - Recombinant human thrombopoietin clinical development. AB - Patients undergoing anticancer therapy are often at risk for developing severe and/or prolonged posttreatment thrombocytopenia. This can be associated with significant bleeding; currently, it is treated with supportive platelet transfusions. Frequent platelet transfusions can cause alloimmunization which requires HLA-matched donors and more frequent blood transfusions, and transmission of both viral and bacterial infections via platelet transfusions remains a concern. Furthermore, thrombocytopenia can mandate a decrease in the dose intensity of cytotoxic therapy by causing either delays or dose reductions in therapy administration. An intervention that reduces the risk or shortens the duration of severe thrombocytopenia would represent an important medical advance. Thrombopoietin (TPO), a naturally occurring, glycosylated polypeptide that was cloned by Genentech in 1994, is capable of inducing differentiation of stem cells into megakaryocytes and accelerating the maturation of megakaryocytes, thereby increasing the platelet count. Recombinant human TPO (rHuTPO) is currently undergoing testing in phase 1 and 2 studies in patients receiving myelosuppressive or myeloablative therapy. For the purposes of illustration, preliminary safety and activity data from one ongoing phase 1 myelosuppression trial (rHuTPO in women with advanced gynecologic malignancies receiving carboplatin) and one ongoing phase 1 myeloablation trial (rHuTPO for peripheral blood progenitor cell mobilization prior to myeloablative chemotherapy for high risk breast cancer) will be presented. PMID- 11012193 TI - FDA licensure of NEUMEGA to prevent severe chemotherapy-induced thrombocytopenia. AB - This paper discusses background information and the body of clinical data that has been accumulated to demonstrate the efficacy and safety of NEUMEGA (recombinant human interleukin 11) when used to prevent severe chemotherapy induced thrombocytopenia and reduce the need for platelet transfusions in patients with nonmyeloid malignancies. NEUMEGA is recommended to be used at a dose of 50 microg/kg s.c. once daily starting the day after chemotherapy ends until a platelet count of 50,000 cells/microl is achieved after the expected nadir. PMID- 11012194 TI - Early Australian clinical studies with pegylated recombinant human megakaryocyte growth and development factor. AB - Pegylated recombinant human megakaryocyte growth and development factor (PEG rHuMGDF, an Mpl ligand) is a truncated form of native thrombopoietin currently undergoing clinical development. A series of studies in Australia have examined the safety and biological activities of PEG-rHuMGDF. Administration of PEG rHuMGDF causes a dose-dependent increase in platelet count but has no effect on white cell count or hematocrit. These platelets are morphologically and functionally normal. When administered following moderately myelosuppressive chemotherapy, PEG-rHuMGDF significantly enhances platelet recovery, although scheduling in relation to chemotherapy may be important in optimizing the full effects. PEG-rHuMGDF mobilizes progenitor cells of multiple hematopoietic lineages, and alters the kinetics of peripheral blood progenitor cell mobilization after chemotherapy and filgrastim. PEG-rHuMGDF is well tolerated and does not cause toxicity similar to that observed with other thrombopoietic cytokines. Numerous studies are underway to help determine the precise role of PEG-rHuMGDF in clinical practice. PMID- 11012195 TI - The use of PEG-rhuMGDF in platelet apheresis. AB - Platelet transfusions are increasingly being used to treat thrombocytopenic conditions ranging from aplastic anemia to that caused by cancer chemotherapy. Although historically whole-blood transfusions were the primary source of platelets for transfusion, random donor platelet concentrates and single-donor apheresis platelets are currently the only products used. The use of these products in the United States varies widely for different medical conditions; for example, surgical patients receive random donor platelet concentrates much more commonly than single-donor apheresis products, while the opposite is true for hematology/oncology patients. The past decade has seen a great change in the type of platelet product prescribed. Whereas random donor platelet concentrates were mostly used in the past, over 60% of the platelets transfused are now obtained from donors by apheresis. A crucial variable in the ability to collect platelets by apheresis is the donor platelet count. With the recent availability of thrombopoietin, there has been considerable interest in using this hematopoietic growth factor to stimulate platelet production in donors. Preliminary studies with the administration to platelet donors of one of the thrombopoietic growth factors, PEG-rHuMGDF, have demonstrated a marked increase in the apheresis yield and no side effects. The PEG-rHuMGDF-mobilized platelets were effective upon transfusion. Whether stimulation of platelet production in donors with thrombopoietic growth factors will become a widely accepted method will depend largely on the safety of this approach for the donor as well as on a number of lesser issues which concern the recipient and blood center. PMID- 11012196 TI - The role of c-Mpl ligands in the expansion of cord blood hematopoietic progenitors. AB - The major limitations to the widespread use of high-dose chemotherapy or radiotherapy followed by autologous or allogeneic transplantation are the scarcity of stem cell donors and the depletion of the autologous stem cell reservoir. Cord blood is a readily available source of stem cells, which, however, might be limited in number. For this reason, up to now, cord blood transplantation has been restricted to children. Therefore, a major goal for experimental and clinical hematology is the identification of mechanisms and conditions that support the expansion of transplantable hematopoietic stem cells. Two systems have been described to identify in vitro these progenitor cell populations in both mice and humans: A) long-term culture-initiating cells (LTC IC), so named because of their ability to support the growth of hemopoietic colonies (colony-forming cell [CFC]) for five to six weeks when cocultured on stromal layers, and B) the generation of hematopoietic progenitors CFC from stroma-free liquid cultures for extended periods of time, which is another indirect evidence for the presence of primitive stem cells. The two systems detect largely overlapping but not identical cell populations of progenitor cells; thus, the identification of the growth factor requirements for the maintenance and amplification of both systems is relevant. The studies presented here demonstrate that CD34+ cord blood cells can be grown in stroma-free liquid cultures for extremely prolonged periods of time (up to six months). During such a period, hemopoietic precursors and committed progenitors belonging to all of the hematopoietic lineages are continuously and massively generated. Such a massive expansion is sustained by an increasingly larger expansion of primitive stem cells (CFU-BI and LTC-IC). The presence of both FL and thrombopoietin (TPO) was necessary and sufficient to support this phenomenon. The addition of KL +/- interleukin 6 (IL-6) does not appear to substantially modify the extent of LTC-IC expansion. FL and TPO appear to be two unique growth factors that preferentially support the self-renewal of primitive stem cells; the additional presence of KL and IL-6 seems to enhance the proliferative potential of at least a subpopulation of daughter stem cells which can undergo at least three differentiation pathways. PMID- 11012197 TI - Opportunities for the use of thrombopoietic growth factors. AB - Highly lineage specific thrombopoietic factors are now available which can produce substantial increments in platelet counts and attenuate the severity of therapy induced thrombocytopenia in preclinical models and early trials in patients. These striking findings are not equivalent to proving clinical benefit to patients, however, and studies are now in progress in a variety of clinical settings. Methodologic issues which could influence the interpretation of studies using thrombopoietin to mitigate thrombocytopenia in patients treated for acute myeloid leukemia and following stem cell transplantation are discussed. Studies in thrombocytopenic patients with myelodysplasia, aplastic anemia, immune thrombocytopenic purpura, as well as selected platelet and stem cell donors, are also of interest. It is unclear, however, whether patients with impaired marrow function can benefit from further exogenous stimulation, and side effects must be very carefully monitored in studies with normal donors. PMID- 11012198 TI - Megakaryocyte and platelet structure in thrombocytopoiesis: the effect of cytokines. AB - This paper presents an overview of selected data which the author considers crucial to an understanding of structure/function relationships of megakaryocytes (MK) and platelets. The observation that platelet territories form within the MK cytoplasm and that, therefore, MK and platelet plasma membranes need not be structurally or antigenically identical is substantiated on the basis of results obtained with a variety of experiments. While the predominant site of MK fragmentation is still debated, it is generally accepted that such terms as "proplatelets," "giant platelets" or "megathrombocytes" refer to MK fragments consisting of more than one platelet territory. It is suggested that such fragments be called "compound" platelets to convey a unifying concept. The terms "young" or "immature" could be reserved for platelets which still contain ribosomes, rough endoplasmic reticulum or other organelles not usually seen in circulating platelets. Finally, the structure changes induced by cytokines, such as interleukin 3 (IL-3), IL-6, IL-11 and thrombopoietin have been illustrated. PMID- 11012199 TI - Measurements of in vivo megakaryocytopoiesis: studies in nonhuman primates and patients. AB - In vivo megakaryocytopoiesis was directly analyzed for megakaryocyte (MK) number and mass, expression of lineage-specific and myeloid differentiation markers, and cell maturation as determined by size, granularity and ploidy. Using a rapid method for multiparameter correlative analysis with three-color flow cytometry (FCM) and a single-argon-ion-laser analyzer, cell DNA in aspirated marrow was stained with 7-amino-actinomycin D, and surface membrane receptors were analyzed with antibodies and cytokines labeled with fluorescein, phycoerythrin and peridinin chlorophyll protein. MKs expressing glycoprotein (GP) IIb/IIIa were enumerated in relation to the nucleated erythroid precursors expressing glycophorin A, and MK diameters were measured by time-of-flight technique. In human marrow (n = 10) the average MK diameter is 37 microm (range: 21 microm for 2N to 56 microm for 64N cells), volume is 26 x 10(3) fL, and MK number is 10 x 10(6)/kg, giving a total MK mass of 26 x 10(10) fL/kg. The modal ploidy is 16N. In essential thrombocythemia patients (n = 10) with a mean platelet count of 907 +/- 23 x 10(6)/L, MK number and volume increased twofold with modal ploidy of 32N, and MK mass fourfold the normal value. After reducing the platelet count to 353 +/- 42 x 10(6)/L with anagrelide therapy, MK number and volume decreased with modal ploidy of 16N, resulting in reduced MK mass by 50%. By contrast, patients with chronic myelogenous leukemia (n = 3) showed an increase in small MKs with a modal ploidy of 8N. In non-human primates, treatment with interleukin 6 or GM-CSF increased MK volume and ploidy with a variable increase in cell number and platelet counts. Treatment with recombinant human MK growth and development factor (n = 6, 5 microg/kg for 28 days) increased platelet count fivefold, MK number fourfold, MK volume twofold and total mass sevenfold. Using three-color FCM, marrow MKs labeled for GPIIb/IIIa and stained for DNA expressed high levels of von Willebrand factor with a high resolution of 2N/4N MKs from the total marrow cells. The expression of myeloid markers including CD36, CD45 and IgG-Fc gammaRII CDw32 correlated directly with increasing cell maturation, concordant with the expression of GPIIb/IIIa and GPIb. Conversely, the expression of HLA-DR declined with maturation. We conclude that pathophysiologic and therapeutic changes in megakaryocytopoiesis in vivo are readily quantified using FCM measurements. PMID- 11012200 TI - Early suppressive effects of chemotherapy on recovery of bone marrow megakaryocyte precursors: possible relationship to platelet recovery. AB - This study utilized a recently developed culture and quantitation system to detect megakaryocyte precursors in CD34+ bone marrow cells from normal donors and breast cancer patients treated with 5-fluorouracil, leucovorin, adriamycin and cyclophosphamide (FLAC). Bone marrow was obtained from patients before and then after their first cycle of FLAC once blood cell counts had recovered. CD34+ cells were isolated and placed in liquid culture with growth factors to stimulate proliferation and lineage commitment. Absorbance values from an enzyme-linked immunosorbent assay were used to quantitate expression of platelet glycoprotein GPIIb/IIIa. There was an increase in absorbance with increasing numbers of cells seeded per culture that was associated with an increase in the number of megakaryocyte lineage cells produced. After 10 days in liquid culture, absorbance values for expression of GPIIb/IIIa from 2,000 normal donor and pre-chemotherapy CD34+ marrow cells were > or = 1.0. Absorbance values from cultures of post chemotherapy CD34+ cells from four patients were similar to values from pre chemotherapy CD34+ cells. In contrast, absorbance values from cultures of post chemotherapy CD34+ cells from two other patients were low (absorbance < 0.5). Low absorbance values for GPIIb/IIIa expression indicate that megakaryocyte production from those CD34+ cells was reduced. Both of those patients developed prolonged thrombocytopenia and platelet nadirs of less than 20,000/microl during FLAC chemotherapy. In contrast, only one out of four patients whose cultures of post-chemotherapy CD34+ cells had absorbance values > or = 1.0 developed platelet nadirs less than 20,000/microl. These results suggest that low platelet nadirs and delayed platelet recovery may be associated with suppressive effects of chemotherapy on recovery of megakaryocyte precursors. PMID- 11012201 TI - The regulated expression of a TATA-less, platelet-specific gene, alphaIIb. AB - The megakaryocyte (MK)-specific integrin, alphaIIb, is the alpha-subunit of the alphaIIb/beta3 complex found on the surface of platelets. This complex is a receptor for fibrinogen and other ligands when platelets are activated. Because the alphaIIb gene is specifically expressed in MKs, this gene was studied as a potential model for MK-specific gene expression. Previous studies have defined some of the important regulatory elements in 912 bp of the immediate 5'-flanking region of this gene. These studies defined several important elements including two GATA-binding elements and an Ets-binding element. Using a primary rat marrow expression system, we demonstrated that one of the GATA-binding elements, -454 bp upstream of the transcriptional start site (GATA454), is critical for expression of the alphaIIb gene. A potential negative regulatory element was found between 100 and -200 bp upstream of both the rat and human alphaIIb genes. The biological basis by which this negative regulatory region effects expression is not well understood. Recent studies have focused on the issue of the molecular basis by which this TATA-less gene is properly transcribed. We found that a GA-rich region approximately 14 bp upstream from the transcriptional start site appears to be a nonconsensus Sp1-binding site that interacts with an Ets-consensus site approximately 20 bp further upstream. These studies provide further evidence of the role of interactions between Ets-like proteins and Sp1 in transcriptional activation when a TATA box is not present in the promoter region of a gene. Based on the presented studies and previous results, a model is proposed for the regulation of expression of the alphaIIb gene. In studies looking at more distal regulatory elements, we have found, using the primary rat marrow expression system, that 2.9 kb of 5'-flanking alphaIIb sequence has as high a level of expression as the 912 bp construct. Whether either of these lengths of 5' flanking region can result in tissue-specific expression in transgenic models is presently being investigated. In addition, while a published report suggests that the two genes alphaIIb and beta3 are physically linked within a 250 kb region of genomic DNA, analysis of yeast artificial chromosome clones and genomic pulsed field gel electrophoresis analysis are consistent with these two genes not being tightly linked and being >1 mb apart, suggesting that these two genes do not form a single, tissue-specific locus. PMID- 11012202 TI - Preclinical biology of megakaryocyte growth and development factor: a summary. AB - Since the discovery of the ligand for the cytokine receptor c-Mpl, much has transpired. The development of this protein has been rapid, and the amount of information available on the effects of this molecule in vitro and in vivo is vast. This paper will highlight some of the major studies and observations which are part of the ongoing pre-clinical development of the megakaryocyte growth and development factor encoding the erythropoietin-like domain of the c-Mpl ligand. A summary of in vitro effects on human cells, as well as the key in vivo observations, are included. This molecule is currently in clinical trials, and the initial results are promising. PMID- 11012203 TI - The role of recombinant interleukin 11 in megakaryocytopoiesis. AB - Recombinant human interleukin 11 (rHuIL-11) is a multifunctional cytokine with activities on a broad range of hematopoietic cells including primitive stem cells and mature progenitor cells. Analysis of rHuIL-11 in vitro has revealed that its hematopoietic activities are predominantly a result of synergistic interactions with other early-acting factors such as IL-3 and Steel factor. Studies indicate that rHuIL-11 acts directly on purified stem and progenitor cell populations and can support the growth of colony forming units-megakaryocyte in these cultures. In normal animals, rHuIL-11 has a potent effect on cells of the megakaryocyte (MK) lineage. Administration of rHuIL-11 results in a two- to threefold increase in circulating platelets, stimulation of bone marrow (BM) and spleen progenitor numbers, and enhanced MK maturation as measured by a shift to higher ploidy values. rHuIL-11 administration in preclinical models of myelosuppression induced by chemotherapy and/or irradiation has shown a reproducible acceleration of platelet recovery and, in some models, enhanced neutrophil and red blood cell recovery. rHuIL-11 has been tested in a non-human primate myelosuppression model using carboplatin. Administration of rHuIL-11 following carboplatin treatment was found to eliminate the period of severe thrombocytopenia (<20,000 platelets/ml) and enhance the recovery of platelets to normal levels (>100,000/ml). Recently, human clinical trials conducted with rHuIL-11 in patients treated with chemotherapy have demonstrated its potent thrombopoietic activity, including improved platelet nadirs, enhanced platelet recovery and a significant decrease in the number of patients who require platelet transfusions. Combined with the preclinical results, these studies confirm that this cytokine will be an effective agent in the treatment of myelosuppression and thrombocytopenia associated with cancer chemotherapy and BM transplantation. PMID- 11012204 TI - Characterization, molecular cloning and expression of megakaryocyte potentiating factor. AB - We examined whether the conditioned media of 64 kinds of cell lines, which have been maintained by a protein-free culture system, could produce megakaryocyte potentiating (Meg-POT) activity. In these cell lines, HPC-Y5, established from human pancreatic cancer, was shown to have the highest level of activity. The megakaryocyte potentiating factor (MPF) was purified from its conditioned medium by a combination of ion-exchange chromatography, gel filtration and reversed phase HPLC. The purified MPF showed Meg-POT activity almost equal to human (Hu) interleukin 6 (IL-6) in the presence of murine IL-3 in a colony-forming assay with mouse bone marrow cells. The molecular weight of MPF was estimated to be 33 kDa by SDS-PAGE. Glycopeptidase F digestion and amino sugar analysis of the factor demonstrated that MPF is a glycoprotein carrying at least one N-linked sugar chain. The N-terminal amino acid sequence of MPF was determined to be Leu Ala-Gly-Glu-Thr-Gly-Gln-Glu-Ala-Ala-Pro-Leu-Asp-Gly-Val-Leu-Ala-Asn. The same or homologous amino acid sequence has not been found in known proteins, demonstrating that MPF may be a novel cytokine which has Meg-POT activity. Then, we isolated HuMPF cDNA from an HPC-Y5 cDNA library using polymerase chain reaction and plaque hybridization methods. The HuMPF cDNA encodes a polypeptide consisting of 622 amino acids, including a signal peptide of 33 amino acids, and with a deduced molecular weight of 68 kDa, although HPC-Y5 cells secrete a 33 kDa form of HuMPF. HuMPF cDNA does not show any significant homology with other known sequences. The cDNA was expressed in COS-7 and Chinese hamster ovary (CHO) cells, and Meg-POT activity was detected in their culture supernatant. The COS-7 cells secreted only a 33 kDa recombinant (r)HuMPF, however, an additional 30 kDa form was detected in the culture medium of CHO cells. The 33 kDa rHuMPF from CHO cells showed Meg-POT activity, but not the purified 30 kDa rHuMPF. The difference in structure and activity between the 33 and 30 kDa forms of HuMPF was ascribed to the existence in the 33 kDa form of the C-terminal 25 amino acid residues. The expression of MPF mRNA was examined by Northern blot analysis using labeled MPF cDNA as a probe. MPF mRNA was detected in HPC-Y5 cells, with an approximate molecular size of 2.4 kb. We also examined the expression of the MPF gene in various human tissues, and the 2.4 kb band was detected only in lung. Then, the immunohistocytochemical analysis and in situ hybridization revealed that MPF producing cells were identified as lung macrophages. MPF may exhibit other biological activities such as regeneration of the lung tissues. PMID- 11012205 TI - Hierarchical structure of human megakaryocyte progenitor cells. AB - Megakaryocytopoiesis is a complex biological process involving a series of cellular events that begins with the pluripotent hematopoietic stem cell and ultimately results in the biogenesis of platelets. A hierarchy of megakaryocyte (MK) progenitor cells has been previously defined based upon studies of in vitro megakaryocytopoiesis. Ontogeny-related changes in MK progenitor cells were analyzed in order to further define this cellular hierarchy. Unifocal colony forming unit-megakaryocyte (CFU-MK)-derived colonies cloned from fetal bone marrow (FBM) formed after fewer days of in vitro culture and were 2.6-fold larger than those colonies cloned from adult bone marrow (ABM). The frequency of CFU-MK derived colonies cloned from ABM was significantly greater. MK colonies, however, cloned from FBM morphologically consisted of both pure MK colonies and mixed colonies containing MKs, in which a core of CD41- cells were surrounded by CD41+ MKs. Large colonies resembling the primitive BFU-MK also were assayed from both FBM and ABM. These BFU-MK-derived colonies appeared after fewer days of incubation when FBM was assayed, compared to ABM, but at a significantly lower frequency. In addition, large unifocal MK colonies consisting of >300 cells (300 1000) appeared from cells cloned from fetal, but not adult, marrow. This type of colony represents a unique type of MK progenitor cell, termed the high proliferative-potential cell-MK. Such colonies represent the progeny of the most primitive human MK progenitor cell identified to date. We also attempted to investigate the process of commitment of stem cells to the MK lineage. We explored the actions of thrombopoietin (TPO) on primitive hematopoietic cells in order to gain an understanding of stem cell commitment. CD34+ Thy-1+ Lin- marrow cells, which are enriched for pluripotent hematopoietic stem cells, were shown to express c-Mpl by the polymerase chain reaction. In addition, TPO alone was capable of inducing CD34+ Thy-1+ Lin- cells after two to three weeks to produce progeny composed entirely of MKs. These studies indicate that TPO has a profound effect on hematopoietic stem cells, and that the hierarchy of MK progenitor cells begins with the pluripotent hematopoietic stem cell. PMID- 11012206 TI - Cloning and functional analysis of erythropoietin-, interleukin-3- and thrombopoietin-inducible genes. AB - The receptor for thrombopoietin (TPO) is a member of the cytokine receptor superfamily. This superfamily also includes the receptors for erythropoietin (EPO), interleukin 3 (IL-3), GM-CSF and several other cytokines. Stimulation of cytokine receptors with their cognate ligands results in the activation of multiple signal transduction pathways and ultimately in the induction of new genes. The cloning of these specific genes provides one approach for analyzing cytokine-specific responses. In the current study, we have designed a strategy for isolating inducible genes. While the strategy has been used to identify EPO specific and IL-3-specific inducible genes, the strategy can be extended to clone TPO-inducible genes. PMID- 11012207 TI - The physiology of platelet production. AB - The production of platelets from the bone marrow megakaryocytes is a well regulated process. Nearly 100 years ago, James Homer Wright described how platelets formed from megakaryocytes and entered the circulation. Subsequent clinical and animal studies have enumerated a number of principles of platelet physiology: the platelet count is constant in any one individual but varies greatly between individuals; an inverse relationship exists between the platelet count and platelet size; the body conserves the mass, not the number, of platelets; and megakaryocyte number, size and ploidy vary in response to changing demands for platelets. With the discovery of thrombopoietin (TPO), a number of additional physiological principles have emerged: TPO takes 24 h to rise maximally and has a maximal half-life of 45 min; TPO levels are inversely and exponentially proportional to the platelet mass; platelets bind and clear TPO from the circulation; and hepatic TPO product on is not altered by changes in the platelet mass. Using these principles, a model for the regulation of platelet production by TPO has been proposed in which the constitutive hepatic TPO produced is removed from the circulation by the platelet mass. Changes in the platelet mass or its ability to clear TPO produce changes in TPO levels resulting in an altered platelet production rate. Using this model, a number of pathological disorders of platelet production, such as essential thrombocythemia and idiopathic thrombocytopenic purpura, are analyzed. PMID- 11012208 TI - Biological roles for the second domain of thrombopoietin. AB - Thrombopoietin (TPO) has been isolated from the plasma of animals in several laboratories as several molecular species which vary in molecular weight from 19 kDa to 35 kDa. Upon biochemical analyses, all of these forms appear to represent proteolytic fragments of TPO which share the same amino-terminal sequences and are therefore truncated at their C-terminal ends. Intact recombinant TPO produced in cell culture is a 70 kDa glycoprotein which contains the full polypeptide backbone encoded by the cDNA sequence. A series of deliberate TPO truncations have been constructed by the introduction of stop codons into the cDNA at various positions. The proteins encoded by the truncated cDNAs have been purified to homogeniety for the purpose of detailed biological and biochemical comparisons of such C-terminally truncated TPO proteins with the full-length intact TPO. These comparisons serve to illuminate the possible biological role(s) of the C-terminal domain, which is so far unique in the family of hematopoietic cytokines. The C terminal domain appears to regulate the specific activity of TPO, to regulate its circulating half-life, and to promote efficient biosynthesis and secretion of the protein. PMID- 11012209 TI - Do the preclinical effects of thrombopoietin correlate with its in vitro properties? AB - In the short time since its cloning, much has been learned of the in vitro properties of thrombopoietin (TPO). In addition to effects on the differentiation of megakaryocytes, TPO has also been shown to stimulate the proliferation of megakaryocytic progenitor cells, colony-forming units-megakaryocytes (CFU-MK), to act in synergy with interleukin 3 or c-kit ligand and erythropoietin (Epo) to stimulate the development of early and the generation of late erythroid progenitor cells, and to affect the rate of entry into the cell cycle and proliferative capacity of hematopoietic stem cells. An important question posed by these observations, for both TPO and for hematopoietic research in general, is whether the in vitro effects of a cytokine are mirrored by its preclinical and clinical biology. The results of recent studies in mice and nonhuman primates will be presented which have attempted to address this issue. In normal animals, TPO increases the numbers of marrow and spleen CFU granulocyte/erythroid/macrophage/megakaryocyte, CFU-MK, CFU-GM and BFU-E, but its effects in the peripheral blood are limited to marked increases in the platelet count. The reason for widespread progenitor cell effects, yet stable leukocyte and erythrocyte blood counts, is likely the predominant regulatory effects of G CSF and Epo; in the absence of elevated levels of the lineage-dominant regulator of each of these cell types, expanded progenitor cell numbers are not translated into increased peripheral blood counts. However, in states of increased blood cell demand such as follows myelosuppressive therapy, elevated levels of Epo and G-CSF allow the effects of TPO on erythropoiesis and myelopoiesis to become manifest. The administration of TPO to myelosuppressed animals is associated with not only greatly expanded hematopoietic progenitor cell recovery, but also improvement in platelet, red cell and leukocyte nadir levels and greatly accelerated recovery of all three cell lineages. These results indicate that the panhematopoietic properties of TPO identified by in vitro culture techniques correlate well with its effects in animals. The results of ongoing clinical trials should soon establish whether these conclusions can be extended to patient care. PMID- 11012210 TI - The role of transcription factor NF-E2 in megakaryocyte maturation and platelet production. AB - To determine the in vivo functions of the transcription factor NF-E2, we have disrupted the gene encoding its hematopoietic-specific p45 subunit in embryonic stem cells and generated knockout mice. These animals have a surprisingly mild erythroid cell abnormality but experience lethal hemorrhage as a result of profound thrombocytopenia. Impaired platelet formation is secondary to a cytoplasmic maturation arrest within megakaryocytes, characterized by a marked reduction in granule numbers. Although the proliferative response to recombinant thrombopoietin is intact, this is not accompanied by correction of the differentiation defect. Absence of the smaller (p18) subunit of NF-E2 does not have similar consequences. These findings implicate target genes of the NF-E2 transcription factor in critical aspects of late megakaryocyte maturation and platelet formation. PMID- 11012211 TI - Dissection of c-Mpl and thrombopoietin function: studies of knockout mice and receptor signal transduction. AB - The physiological roles and mechanisms of action of thrombopoietin (TPO) and its receptor c-Mpl have been studied through the analysis of mice genetically deficient in these molecules, as well as through the dissection of signaling events utilizing chimeric receptors. The evidence clearly demonstrates that the TPO/c-Mpl system provides dominant control in the regulation of megakaryocytopoiesis. The signaling mechanisms that underlie this process appear to be similar to those noted with other members of the hematopoietic cytokine and cytokine receptor families. PMID- 11012212 TI - Studies of the c-Mpl thrombopoietin receptor through gene disruption and activation. AB - The c-mpl gene encodes a receptor for thrombopoietin (TPO), a cytokine that potently stimulates megakaryocytopoiesis. To study the mechanisms of c-Mpl activation, we generated constitutively active receptor mutants. Substitution of cysteine residues into a dimer interface homology domain of c-Mpl forced ligand independent homodimerization and constitutive receptor activation. In factor dependent cells, mutant receptors induced autonomous growth and tumorigenicity. The receptors were constitutively phosphorylated in these cells, as were signal transduction molecules implicated in Mpl function. These data suggest that the normal process of ligand-induced Mpl activation involves receptor homodimerization and that mutated forms of the cellular mpl gene can contribute to tumorigenicity. We have also examined the biological role of c-Mpl in mpl deficient mice generated via homologous recombination in embryonic stem cells. Homozygous mutant animals were deficient in megakaryocytes and severely thrombocytopenic. Mature cells from all other hemopoietic lineages were unaffected. Bone marrow cells from mpl-/- mice were incapable of binding to TPO or responding to the cytokine in clonogenic assays, and further displayed a marked deficiency in progenitor cells capable of megakaryocyte colony formation in response to other stimuli. Moreover, total progenitor numbers were also deficient and included significant reductions in colony-forming cells of multiple hemopoietic lineages. Unexpectedly, the numbers of progenitor cells of all lineages were not perturbed in mid-gestation mpl-/- fetal liver. Our analyses suggest an indispensable role for c-Mpl in megakaryocyte development and reveal that the function of TPO and its receptor is not confined solely to activities in megakaryocytopoiesis. PMID- 11012213 TI - Physiologic role of TPO in thrombopoiesis. AB - Recombinant human thrombopoietin (rHuTPO) serves as a megakaryocyte colony stimulating factor and predominantly acts on GPIIb/IIIa+ rat late megakaryocyte progenitor cells, colony forming units-megakaryocyte (CFU-MK). The GPIIb/IIIa+ fraction of CFU-MK differentiates into mature megakaryocytes and further into proplatelets in liquid culture containing rHuTPO. rHuTPO stimulates cultured megakaryocytes generated from rat GPIIb/IIIa+ CFU-MK to enhance proplatelet formation and to increase megakaryocyte size. rHuTPO also induces a big size of megakaryocyte colonies from human cord blood CD34+ cells. rHuTPO does not cause aggregation of platelets from normal mice and mice made thrombocytotic by consecutive administration of rHuTPO, but preincubation with rHuTPO enhances adenosine diphosphate-induced aggregation, suggesting that platelets induced by rHuTPO administration may have a normal function. Administration of rHuTPO to normal mice daily for five days causes a dose-dependent thrombocytosis. On the other hand, rHuTPO induces a significant decrease in hemoglobin concentration and does not affect white blood cell counts. rHuTPO increases the size and number of marrow megakaryocytes and the number of marrow CFU-MK, and also influences the development of other hematopoietic progenitor cells. The effects of rHuTPO on thrombocytopenia associated with myelosuppression were examined in animal models. Following treatment with mitomycin C, mice received daily injections of various doses of rHuTPO. rHuTPO reduced the severity of thrombocytopenia, accelerated the recovery of platelets and improved neutropenia. Similar therapeutic efficacy was observed in cynomolgus monkeys treated with nimustine. These results suggest the clinical usefulness of rHuTPO for the treatment of thrombocytopenia. PMID- 11012214 TI - Protein characteristics of thrombopoietin. AB - Thrombopoietin (TPO) was purified from irradiated thrombocytopenic rat plasma. In the process of purification, some biochemical and biological characteristics were investigated. Rat plasma TPO was extremely hydrophobic and exhibited multiple peaks of activity on gel filtration. Both the low and high molecular weight fractions were separately subjected to further purification. Consequently, a rat TPO cDNA was cloned based on the amino acid sequences of purified rat plasma TPO. It revealed that each final purified rat plasma TPO was not a full-length form. In addition, rat hepatocytes and three rat hepatoma cell lines were found to produce rat TPO. Each native TPO derived from cultured cells was also partially purified, and hepatocyte-derived TPOs were shown to be heterogeneous in molecular weight. To study the structure of TPO, various recombinant TPO molecules were generated. Two disulfide bonds (Cys7-Cys151 and Cys29-Cys85) located in the N terminal domain of TPO have an important effect on its biological activity. The human TPO muteins, sequentially deleted from the C-terminal, were expressed in COS-1 cells. TPO (1-151) was active, but TPO (1-150), which lacks Cys151, did not exhibit TPO activity. These findings indicate that the region essential for TPO activity is the N-terminal domain, which contains two disulfide bonds. Although the role(s) of the C-terminal domain is not clear at present, the potential N glycosylation in the C-terminal domain is not directly required for exhibiting TPO activity. PMID- 11012215 TI - Gene expression and transcriptional regulation of thrombopoietin. AB - Thrombopoietin (TPO) is predominantly expressed in the liver among various tissues that express TPO transcripts. To investigate the transcriptional regulation of the human TPO gene in the liver, we determined the major transcription initiation site by means of 5'-RACE and Northern blotting. From these analyses, we concluded that TPO gene transcription started at various points, and the transcription initiation sites of the human TPO gene were localized downstream, close to a point we determined by S1 nuclease mapping. The human TPO promoter region contains consensus sequences of GATA, Evi-1, and Ets binding sites. We used the hepatocellular carcinoma cell line, HepG2, that expresses TPO mRNA to analyze its promoter activity by transfecting various reporter plasmids containing a sequentially 5'-deleted human TPO promoter. Although GATA binding factors increased the promoter activity, their effect was independent of the GATA binding consensus sequence. On the other hand, Evi-1 did not affect transcription. Moreover, we defined the core promoter region, in which an Ets binding consensus sequence was located. The deletion or mutation of the Ets binding site resulted in a loss of the promoter activity. These results suggested that TPO is regulated by the Ets family of transcription factors. PMID- 11012217 TI - Megakaryocytopoiesis in vitro: from the stem cells' perspective. AB - Megakaryocytopoiesis is a complex network regulated by different megakaryocyte (MK)-stimulating factors (i.e., thrombopoietin [TPO], stem cell factor [SCF], interleukin 3 [IL-3], IL-6, IL-11 and GM-CSF). Although all of these factors can affect human and murine megakaryocytopoiesis at different levels of MK development, the effect on very primitive hematopoietic stem cells (HSC) is not well understood. We have further characterized the in vitro biological activity of recombinant murine TPO, SCF and IL-3 on the maturation and proliferation of MK progenitors from different murine primitive hematopoietic cells in a fibrin clot system under serum-free conditions. Neither TPO nor SCF alone induced MK colony formation (CFU-MK) from Lin- Sca+ cells. However, isolated large and mature MKs were observed in the presence of TPO. In contrast, IL-3 exerted a potent effect on CFU-MK formation from Lin- Sca+ cells. On this population of HSC, a significant increase of large MK colonies with mature MK were obtained under those conditions in which TPO was combined with IL-3 or SCF plus IL-3. Similar results were obtained with murine bone marrow cells enriched by primitive progenitors from day 3 post-5-fluorouracil treated mice (5-FUBMC). In contrast, TPO-sensitive precursors were detected in fetal liver cells (FLC). These cells differentiate and proliferate to MK progenitors in the presence of TPO. A significant increase in the number of CFU-MK was induced when TPO was combined with either IL-3 or SCF. On these populations of primitive hematopoietic progenitors, IL-3 induced both the proliferation and differentiation of MK progenitors. Because erythropoietin and TPO share similarities between their molecules and their receptors, we studied whether these growth factors may modulate megakaryocytopoiesis from FLC. Flow cytometry analysis of FLC expressing erythroid markers demonstrated that these cells expressed c-Mpl receptor. In our in vitro studies, although EPO by itself did not induce MK colonies from FLC, it enhanced the proliferative activity of TPO. High ploidy and proplatelet-shedding MK were observed in Lin- Sca+ cells, 5-FUBMC and FLC stimulated with TPO alone or in combination with other MK-stimulating factors. Based on these observations, we propose that TPO, IL-3 and SCF constitute early MK-acting factors with differential proliferative and differentiative activities on murine stem cells. TPO by itself does not appear to be involved in the proliferation of MK progenitors from bone marrow HSC. TPO appears to induce in these cells the commitment toward MK differentiation. However, this growth factor may enhance the proliferative activity of IL-3. IL-3 is an early MK-stimulating factor able to induce in vitro the proliferation and differentiation of MK progenitors from HSC. PMID- 11012216 TI - Cytokine-induced alteration of platelet and hemostatic function. AB - A number of nonplatelet-specific cytokines that augment platelet recovery following chemo/radiotherapy have been described. The members of the interleukin 6 (IL-6) family have properties that influence the hematopoietic system beyond their modest thrombocytopoietic effects. Studies performed in a canine model with IL-6 have shown that this factor augments plasma fibrinogen and von Willebrand factor (vWf) concentrations and decreases the level of free protein S. IL-6 appears to decrease the bleeding time in thrombocytopenic dogs, although this effect does not seem to be due to a direct influence of the factor on endothelial vWf or tissue factor production. The factor does not directly alter platelet function in vitro, but when administered to dogs, it increases the sensitivity of the platelets to activation by thrombin. Normal platelets injected into IL-6 treated dogs, and platelets from IL-6-treated dogs injected into normal animals, survive normally. Following injection of either IL-6 or the more specific thrombocytopoietic cytokine thrombopoietin (TPO), IL-6 increases platelet responsiveness to thrombin-induced activation to a greater extent than does TPO. The data show that IL-6 has certain properties that might be construed as prohemostatic, and these properties may prove to be useful clinically. PMID- 11012218 TI - Thrombopoietin, a direct stimulator of viability and multilineage growth of primitive bone marrow progenitor cells. AB - Thrombopoietin (TPO), the ligand for c-mpl, has recently been demonstrated to be the primary regulator of megakaryocytopoiesis and platelet production. In addition, several studies have demonstrated that c-mpl is expressed on hematopoietic cell populations highly enriched in primitive progenitor cells. Here we summarize and discuss recent studies from our laboratory, as well as others, demonstrating that TPO has effects on primitive hematopoietic progenitor cells. When acting alone, TPO stimulates little or no growth, but promotes viability and suppresses apoptosis of murine multipotent (Lin- Sca-1+) bone marrow progenitor cells in vitro. In addition, TPO directly and potently synergizes with other early acting cytokines (kit ligand, flt3 ligand and interleukin 3) to promote multilineage growth of the same progenitor cell population. Although it remains to be established whether TPO also acts on the long-term reconstituting pluripotent stem cells, these studies combined with progenitor cell studies in c-mpl-deficient mice, suggest that TPO, in addition to its key role in platelet production, might also have an important impact on early hematopoiesis. PMID- 11012219 TI - A new transgenic mouse model for the study of cell cycle control in megakaryocytes. AB - During the development of the megakaryocytic lineage, the megakaryoblasts give rise to megakaryocytes which undergo repeated S phases in the absence of cytokinesis (endomitosis). The cellular oncogene myc plays a central role in the proliferation and differentiation of several cell types. In a previous study, we generated transgenic mice carrying c-myc fused to the estrogen receptor under the control of the platelet factor four (PF4) megakaryocyte-specific promoter. The bone marrow of female transgenic mice, but not of male mice, displayed increased megakaryopoiesis. Here we report that beta-estradiol-induced activation of c-myc in cultured bone marrow cells derived from male or female transgenic mice resulted in prolonged survival of the cells in vitro. Addition of a cocktail of hemopoietic growth factors to beta-estradiol-treated cells, including interleukin 6 (IL-6), IL-3 and stem cell factor further improved the survival time in culture and increased the percentage of large mature cells, but did not result in immortalization. The majority of these PF4-expressing cells, however, did not reach the differentiation stage at which acetylcholinesterase is expressed and did not appear as large megakaryocytes. We conclude that cultured megakaryocytes overexpressing myc are induced to proliferate, but have a limited potential to fully differentiate. Under these conditions, cyclin D3 was downregulated while the level of cyclin A was slightly upregulated. PMID- 11012220 TI - The platelet glycoprotein Ib-V-IX system: regulation of gene expression. AB - Platelet glycoproteins (GPs) Ib-V-IX form the surface receptor for von Willebrand factor, and this receptor-ligand interaction mediates the shear-dependent adhesion of platelets to damaged arterial vessel walls. The receptor is a multicomponent structure consisting of four distinct polypeptides (heterodimeric GPIb: Ib alpha-Mr143k and Ib beta-Mr22k; GPV-Mr83k; GPIX-Mr20k), and each of the four cDNAs and genes has been cloned and characterized. The genes appear to have evolved from a common progenitor genomic sequence related to that encoding GPIX. They share simple structures with few introns and possess common consensus regulatory sequences (GATA, ets, Sp-1) in their 5' flanks. Both the GPIb alpha and the GPIX promoters have been analyzed by transfection of reporter constructs into hematopoietic and nonhematopoietic cells. The promoters function in a tissue specific fashion, and gel shift and mutational analyses indicate that GATA and ets sequences regulate activity. In the case of the GPIX promoter, footprints confirm the role of the ets-related consensus region. Recent studies of GPIb beta transcriptional regulation suggest that an aberrant polyadenylation signal, located in the 3' end of the gene immediately upstream of the GPIb beta gene, allows in vitro expression of a rare extended fusion transcript encoding both the upstream protein and GPIb beta. Little detailed information is available in regard to expression of the GPV gene. In summary, the genes of the GPIb-V-IX system display features of other megakaryocyte/platelet genes, but the unique regulatory events that direct the selective expression of these genes in megakaryocytes remain to be defined. PMID- 11012221 TI - Hematopoietic differentiation of embryonic stem cells: an in vitro model to study gene regulation during megakaryocytopoiesis. AB - We are interested in the regulation of the tissue specificity of the megakaryocyte-specific platelet glycoprotein IIb gene. The murine embryonic stem (ES) cells are able to differentiate into erythroid, mast and granulomonocytic cells in appropriate culture conditions. Our goal is to optimize the production of myeloid cells including megakaryocytes (MKs) by ES cells. We have found that coculture with MS-5 stromal cells and the presence of a cocktail of hematopoietic growth factors (HGFs) [stem cell factor, interleukin 3 (IL-3), IL-6, IL-11, G-CSF and erythropoietin] had a high synergistic activity on differentiation of ES cells into pure and MK-containing myeloid colonies from day 12 embryoid bodies. Thrombopoietin increased the number of MKs only when added to the HGF cocktail in the presence of MS-5 cells. Interestingly, many MKs exhibited a "hairy" appearance evocative of pseudopodial proplatelet formation. Expression of genes specific for the megakaryocytic lineage, GPIIb, PF4, mpl and GPIIIa, was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) during differentiation of ES cells, and their relative time course was evaluated. This demonstrates that optimized culture conditions for the differentiation of ES cells into the MK lineage provide a useful tool for the study of the regulation of expression of genes during megakaryocytopoiesis. PMID- 11012222 TI - Dissecting megakaryocytopoiesis in vivo with toxigenes. AB - The genetic programs that regulate the commitment of a totipotent stem cell to the megakaryocytic lineage remain poorly defined and require appropriate in vivo models. Using a cell-specific obliteration technique, a transgenic mouse model was produced where perturbations of megakaryocytopoiesis and platelet production may be induced on demand. This was achieved by targeting the expression of the herpes virus thymidine kinase (HSV-tk) to megakaryocytes using the regulatory regions of the gene coding for the alphaIIb gene, an early marker of megakaryocytopoiesis, which encodes the alpha subunit of the platelet integrin alphaIIb beta3. The HSV-tk gene is not toxic by itself, but sensitizes the target cell to the effect of ganciclovir (GCV), leading to the inhibition of DNA synthesis in dividing cells. The programmed eradication of the megakaryocytic lineage was induced by treating transgenic mice bearing the hybrid construct (alphaIIb-tk) with GCV. After 10 days of treatment, the platelet number was reduced by greater than 96.5% and megakaryocytes were not detectable in the bone marrow (BM). After discontinuing GCV, BM was repopulated with megakaryocytes, and the platelet count was restored within seven days. The recovery was accelerated by the administration of interleukin 11. Prolonged GCV treatment induced erythropenia in the transgenic mice. Assays of myeloid progenitor cells in vitro demonstrated that the transgene was expressed in early erythro-megakaryocytic bipotent progenitor cells. The reversibility and facility of this system provide a powerful model to determine both the critical events in megakaryocytic and erythroid lineage development, and for evaluating the precise role that platelets play in the pathogenesis of a number of vascular occlusive disorders. PMID- 11012223 TI - Methodologic approaches for investigating human megakaryocyte development at a molecular level. AB - Rapid advances in defining the extracellular regulators of megakaryocyte (MK) development have heightened interest in defining the biochemical and molecular mechanisms of action of these cytokines. The recent development of molecular micromethodologies such as in situ hybridization, the polymerase chain reaction, and the use of antisense oligodeoxynucleotides now make such studies possible in normal cells. To illustrate the utility of these methods, data gathered using these methods on developing normal human hematopoietic cells and MKs is presented. As might be expected, the results obtained demonstrate that growth factors have complex time and concentration effects on gene expression in morphologically recognizable human MKs. They also suggest that a more complete understanding of normal MK development at the molecular level will soon be possible. PMID- 11012224 TI - Fibronectin isoforms in megakaryocytes. AB - Our studies have shown that megakaryocytes (MK) can synthesize fibronectin (FN) and alternatively spliced fibronectin, FN EIIIB. FN EIIIB is primarily present in embryonic, proliferating and migrating cells, and thought to be important for cell maturation. MK, but not nonmegakaryocytic bone marrow cells, contain FN EIIIB and thus, MK and platelets are among a small number of adult cells and tissues that synthesize and contain FN EIIIB. Thrombin can induce the secretion of general FN, but does not cause the secretion of FN EIIIB into the medium. Analysis of immunostained cells by confocal microscopy revealed that both general FN and FN EIIIB accumulated on the MK surface following thrombin treatment. Thus, FN EIIIB can be released only to be bound to the MK surface. The expression of FN EIIIB on the MK surface may have a unique role in MK migration and maturation. PMID- 11012225 TI - Regulation of expression of megakaryocyte and platelet proteoglycans. AB - The existence of proteoglycans in hematopoietic cells has been recognized for many years. However, elucidation of the structure and function of these molecules has only begun to be explored in recent years. This paper reviews the current status of knowledge of the structure, function and metabolism of the serglycin proteoglycan in megakaryocytes and megakaryocytic tumor cells. We have identified complex metabolic patterns of the serglycin proteoglycan in terms of regulation of overall hydrodynamic size, glycosaminoglycan chain length and disaccharide composition, and processing of the core protein in control cells or in the presence of phorbol 12-myristate 13-acetate or dimethylsulfoxide. We are currently studying the regulation of synthesis of this protein by analysis of promoter constructs in megakaryocytic and non-megakaryocytic hematopoietic cells. We have also tentatively identified a second proteoglycan, betaglycan, which is known also as the Type III transforming growth factor beta receptor. We have identified this molecule in human erythroleukemia and CHRF 288-11 cells by the presence of characteristic core proteins between 92-120 kDa, by its ability to adhere to Octyl Sepharose and by detection of mRNA. We hope to apply studies of proteoglycan metabolism in these cells to understanding the development of alpha granules and membrane elements in megakaryocytes. PMID- 11012226 TI - Transcriptional regulation in megakaryocytes: the thrombopoietin receptor gene as a model. AB - The MPL gene codes for the thrombopoietin receptor, whose ligand specifically controls megakaryocytic differentiation. In order to understand the molecular basis for the megakaryocyte-specific expression of MPL, we analyzed the regulatory elements of this gene. Two regions are hypersensitive to DNase I in nuclei of cells that express MPL: the promoter and a portion of intron 6. The latter behaves as a chromatin-dependent enhancer. A 200 bp fragment of the promoter is sufficient for high-level specific expression. This fragment can bind several transacting factors in vitro, including GATA-1 and members of the Ets family. GATA-1 binds with low affinity to a unique GATA motif at -70 in the MPL promoter, and destruction of this site yields only a modest decrease in expression level in human erythroleukemia (HEL) cells. Ets proteins also bind with low affinity to two sites. One is located at position -15 and its destruction reduces expression to 50%; the other is located immediately downstream of the GATA motif and plays a crucial role in expression of the promoter in HEL cells, as its inactivation reduces expression to 15%. This study indicates a molecular basis for the coregulation of markers of megakaryocyte differentiation. Finally, we describe other nuclear factor binding sites that may be involved in the cell-type-specific expression of MPL. PMID- 11012227 TI - Platelet immunoregulatory factors. AB - A number of soluble and membrane-associated proteins are known to mediate platelet:leukocyte interactions. Platelet-derived factors that have attracted the most attention to date include transforming growth factor beta, interleukin 1 and platelet factor 4. Recently, we have uncovered another protein within platelets that has leukocyte modulatory activity. It was previously characterized as an endometrial glycoprotein named placental protein 14 (PP14) with suppressive effects upon lymphocyte proliferation, pro-inflammatory cytokine production and natural killer cell function. The "hematopoietic" PP14 derived from cells of the megakaryocytic lineage shares this immunosuppressive property, as evaluated by two-way mixed lymphocyte cultures. Interestingly, two alternatively spliced hematopoietic PP14 mRNAs have been cloned which differ in their encoded proteins. Cell-free translation and transfection analyses have verified the translatability of both PP14 mRNA species and allowed for the analysis of their glycosylation properties. PP14, a member of the lipocalin structural superfamily of proteins, now offers an intriguing new link between the coagulation and immune systems. PMID- 11012228 TI - In vivo biological effects of various forms of thrombopoietin in a murine model of transient pancytopenia. AB - Thrombopoietin (TPO) is the natural regulator of platelet production in the bone marrow of mammals. This cytokine also seems to play an important role in the development of the erythroid lineage when recovering from anemic conditions. Here we study the effects of various TPO molecules on the recovery of hematopoietic lineages in a mouse model of pancytopenia. Based on previous animal experimentation and clinical experience with other hematopoietic cytokines, we found that daily dosing with TPO augmented the recovery of both the megakaryocyte and erythroid lineages in a mouse model of pancytopenia. However, further experiments showed that no benefit was gained by using more than a single dose of recombinant murine (rm)TPO(335) given 24 h after the initiation of the myelosuppressive treatment. This response to a single dose of rmTPO(335) is dose dependent. However, the response was attenuated when a truncated, short half-life TPO molecule (rmTPO[153]) was used. Increasing the half-life of the molecule with 10 kDa polyethylene glycol (PEG) does not improve the response. Only when larger PEG molecules (20 kDa or 40 kDa) are linked to the rmTPO(153) is the response to single doses restored to the level of the full-length molecule. These data suggest that, unlike our experience with other cytokines, the commitment of progenitors to a megakaryocytic cell line is accomplished by a single short exposure to TPO. PMID- 11012229 TI - The clinical development of recombinant human interleukin 11 (NEUMEGA rhIL-11 growth factor). AB - Completed phase I and II studies of recombinant human interleukin 11 (rhIL-11) demonstrate its potential as a treatment for chemotherapy-induced thrombocytopenia. In a phase I study, 16 women with breast cancer received rhIL 11 (10, 25, 50, 75 or 100 microg/kg s.c. once daily) before and during cycles of moderately dose-intensive chemotherapy. Platelet counts increased in all patients before chemotherapy. During chemotherapy, the mean platelet count nadirs were 67,000 cells/microl (rhIL-11 10 microg/kg) and greater than 150,000 cells/microl (25, 50 and 75 microg/kg). Thus, doses of 25 microg/kg and higher appeared to prevent chemotherapy-induced thrombocytopenia in this study. In a randomized, placebo-controlled study, rhIL-11 (50 microg/kg) prevented the need for platelet transfusions during a subsequent chemotherapy cycle in patients who had already experienced severe chemotherapy-induced thrombocytopenia. Among 82 evaluable patients, 8 (30%) of 27 patients administered rhIL-11 50 microg/kg avoided platelet transfusions versus one (4%) of 28 who received placebo (p < 0.05). rhIL 11-treated patients received approximately two-thirds the number of platelet transfusions that placebo-treated patients received. The median duration of thrombocytopenia (<50,000 cells/microl) was seven days in rhIL-11-treated patients compared to 10 days among patients given placebo. This is the first study in which patients with a history of severe chemotherapy-induced thrombocytopenia who were receiving a variety of chemotherapy regimens have been shown to avoid platelet transfusions following the administration of a thrombopoietic growth factor. This activity of rhIL-11, and the demonstration in preclinical models that it ameliorates chemotherapy-induced mucositis, have promoted its further clinical development as a supportive therapy in patients receiving chemotherapy. PMID- 11012230 TI - The problem of thrombocytopenia after hematopoietic stem cell transplantation. AB - Thrombocytopenia after hematopoietic stem cell transplantation (HSCT) is associated with an increased risk of bleeding and utilization of significant resources. This review presents an analysis of risk factors associated with delayed platelet engraftment. The retrospective analysis included 1,468 recipients of autologous or allogeneic transplants treated between January 1, 1990 and July 1, 1995. Risk factors associated with delayed platelet engraftment after autologous HSCT included use of marrow rather than peripheral blood as the source of stem cells, being transplanted for acute myeloid leukemia rather than other diseases, positive patient serology for cytomegalovirus and the presence of infection post-transplant before engraftment. Risk factors associated with delayed platelet engraftment after allogeneic marrow transplantation included unrelated as opposed to related donor transplants, being transplanted for diseases other than chronic myelogenous leukemia, increased age, onset of acute graft-versus-host disease (AGVHD), male gender, the administration of methotrexate for GVHD prophylaxis and the presence of infection before engraftment. Delayed platelet recovery is associated with decreased survival after both autologous and allogeneic transplants. Management of delayed platelet recovery by transfusion of blood products requires significant medical resources and is of some risk to the patients. Further development of new strategies may safely reduce the need for blood products. These include peripheral blood stem cell transplants (allogeneic and autologous), new algorithms for administering routine platelet transfusions and investigative biological agents for stimulating megakaryocytopoiesis. Further studies may elucidate the cause of increased platelet consumption associated with infection and GVHD. PMID- 11012231 TI - The role of platelet growth factors in cancer therapy. AB - The use of myeloid growth factors has markedly reduced the complications of chemotherapy-induced neutropenia, however, abrogation of severe thrombocytopenia remains a major clinical problem. Platelet transfusions remain the standard method of preventing or treating thrombocytopenia but are associated with a variety of complications and are a limited resource. A number of cytokines have been clinically investigated for their thrombopoietic activity, the most promising of which is the recently cloned ligand to the hematopoietic growth factor receptor, c-Mpl. The c-Mpl ligand, also referred to as thrombopoietin, megakaryocyte growth and development factor (MGDF) and megapoietin, is a potent lineage-specific agent that promotes growth and maturation of megakaryocytes and their progenitors. It holds promise for clinical use in the treatment of iatrogenic or disease-associated bone marrow failure states and possibly in syndromes of excessive platelet consumption. Early clinical trials assessing the safety and activity of recombinant human MGDF are now underway. PMID- 11012233 TI - Fluoroquinolone-resistance in Neisseria gonorrhoeae, Hawaii, 1999, and decreased susceptibility to azithromycin in N. gonorrhoeae, Missouri, 1999. AB - In 1999, 360,076 cases of gonorrhea were reported in the United States (1). Gonorrhea is a major cause of pelvic inflammatory disease, often leading to ectopic pregnancy and infertility, and it can facilitate human immunodeficiency virus (HIV) transmission (2). During the 1980s, resistance to penicillin and tetracycline among gonococcal isolates became widespread; as a result, CDC recommended that other antimicrobial agents be used to treat gonorrhea. This report summarizes investigations of an increase in fluoroquinolone-resistant Neisseria gonorrhoeae in Hawaii and of a cluster of N. gonorrhoeae infections with decreased susceptibility to azithromycin in Missouri. PMID- 11012235 TI - What determines benefit from hearing aids? PMID- 11012234 TI - State-specific changes in singleton preterm births among black and white women- United States, 1990 and 1997. AB - National infant mortality rates among non-Hispanic black women are twice those of non-Hispanic white women (1). Nearly two-thirds of this disparity is attributable to a higher rate of preterm delivery (PTD) (i.e., < or = 37 weeks' gestation) among blacks (2). To investigate state-specific changes in PTD rates among blacks and whites, natality data for 1990 and 1997 were analyzed from 50 states and the District of Columbia (DC). These data indicated that, although the PTD rate was twice as high among blacks than among whites, the disparity decreased as the result of an increase in preterm births among whites and a decrease among blacks (3). PMID- 11012236 TI - Distortion product otoacoustic emissions to single and simultaneous tone pairs. AB - Distortion product otoacoustic emissions (DPOAEs) evoked by single tone pairs and three simultaneous tone pairs were recorded in 60 normal-hearing adult ears. The purpose was to replicate a previous study using the commercially available probe assembly of the Grason Stadler GSI 60 and including ear of presentation in the statistical analysis along with frequency and condition. DPOAE levels were comparable between ears and conditions, although differences among frequencies were found. Noise levels were comparable between ears and tended to increase with increases in frequency for both conditions. The latter trend was not noted with the previous study and may be due to differences in the probe assembly and/or test environment. Further, noise levels were significantly greater at 2,000 and 8,000 Hz in the simultaneous condition. Caution should be exercised when interpreting results in the simultaneous condition because not all frequencies may have optimal signal-to-noise ratios when the test is terminated. PMID- 11012237 TI - Effects of electrode location on speech recognition with the Nucleus-22 cochlear implant. AB - Speech recognition performance was measured as a function of electrode in two experiments with the Nucleus-22 cochlear implant using 4-electrode SPEAK speech processors. In experiment 1, the four stimulated electrode pairs were shifted in 0.75-mm steps over 3 mm in the apical-basal direction. In experiment 2, the four electrodes were closely spaced and positioned apically, medially, or basally. An additional condition spaced the four electrodes as widely as possible. In experiment 1, City University of New York sentence scores showed a significant decrease in performance as the electrodes were shifted basally; no other speech measures showed a significant change with electrode location. For experiment 2, all scores were the best with the processor that had the electrodes spaced as widely as possible. In both experiments, all 4-electrode SPEAK processors produced significantly poorer speech recognition than the subject's own 20 electrode processor. These results indicate that the location of electrodes is an important factor in implant performance. PMID- 11012238 TI - Hearing aid benefit in patients with high-frequency hearing loss. AB - Patients with hearing loss limited to frequencies above 2 kHz are often considered borderline candidates for hearing aids. In this study, we used the Profile of Hearing Aid Benefit to access 134 patients' perceived benefit with a variety of linear hearing aids, some more capable than others at achieving prescribed frequency gain targets. We also sought to explore various audiologic and subject factors that might have led patients to report different degrees of success or failure with their hearing aids. Results demonstrate that subjects with hearing loss limited to frequencies above 2 kHz benefit significantly from amplification. However, the amount of benefit reported is mostly unrelated to the hearing aid gain and frequency response. Of numerous audiologic and demographic factors explored in the present study, the number of hours of hearing aid use per day turned out to be the most important single factor that was significantly related to the amount of reported hearing aid benefit. However, the predictive value of knowing how many hours per day subjects wore their aids, or any other combination of factors explored, was quite limited and only accounted for a small amount of the variability observed in user benefit. PMID- 11012239 TI - Development and standardization of SCAN-C Test for Auditory Processing Disorders in Children. AB - This paper reports on the development and standardization of SCAN-C: Test for Auditory Processing Disorders in Children-Revised. The revisions include new test instructions that have been reworded to make them easier for young children, aural stimuli presented on a compact disc, a revision of the Competing Words subtest, and the addition of a Competing Sentences subtest. Normative data on the new test were obtained on 650 children age 5 years, 0 months, to 11 years, 11 months. Analysis of new standardization data revealed systematic improvement in performance with increasing age. A new method of calculating the composite standard score gives equal weighting to each subtest of SCAN-C. Subtest test retest reliability was substantially improved over the original SCAN, with SCAN-C correlations ranging from .65 to .82 for 5 to 7 year olds. Concurrent validity tests found that SCAN-C test results can be viewed with the same confidence as SCAN. PMID- 11012240 TI - Early-onset sensorineural hearing loss in a child with Turner syndrome. AB - Turner syndrome is among the more common but less familiar syndromes that include sensorineural hearing loss and middle ear disease. This article provides a review of the syndrome, an illustrative case, and a review of specific issues relevant to audiologic management of patients with Turner syndrome. PMID- 11012241 TI - Effects of selective serotonin reuptake inhibitors on auditory processing: case study. AB - Auditory sensitivity and processing ability were evaluated in a patient who suffered from hyperacusis, difficulty understanding speech, withdrawn depression, lethargy, and hypersensitivity to touch, pressure, and light. Treatment with fluvoxamine and fluoxetine (selective serotonin reuptake inhibitors) reversibly alleviated complaints. Testing while medicated and unmedicated (after voluntary withdrawal from medication for several weeks) revealed no difference in pure-tone thresholds, speech thresholds, word recognition scores, tympanograms, or acoustic reflex thresholds. Medicated SCAN-A (a screening test for central auditory processing disorders) results were normal, and unmedicated results were abnormal. Unmedicated transient otoacoustic emissions and auditory brainstem response waves I, III, and V were significantly larger bilaterally. Uncomfortable loudness levels indicated greater tolerance during the medicated condition. Central processing and vigilance were evaluated with analog-synthesized three-formant consonant-vowel syllables. While medicated, responses to stimuli at each ear revealed well-defined, labeling crossovers of about 90 msec. Vowel identification matched normal subject responses; labeling of /gE/jE/ and /bE/wE/ continua was well defined but all crossover points differed from normals (p < .0001). During unmedicated testing, responses to /gE/jE/ began at medicated levels but approached chance levels for the entire continuum within 10 min; labeling of /bE/wE/ was consistent with medicated responses throughout with earlier than normal crossover points. PMID- 11012242 TI - A vigorous protest. PMID- 11012244 TI - Dendritic cells: multi-lineal and multi-functional. PMID- 11012243 TI - A high-resolution view of NK-cell receptors: structure and function. PMID- 11012245 TI - The biology of recognitive repertoires. PMID- 11012246 TI - British Society for Matrix Biology Meeting. Aberdeen, 6-7 September 1999. Abstracts. PMID- 11012247 TI - Marked cognitive decline in a premorbidly very bright girl after onset of psychosis. PMID- 11012248 TI - Early onset schizophrenia in a patient with premature birth, germinal matrix hemorrhage and periventricular leukomalacia. PMID- 11012249 TI - Retraction of several recent papers from Dr. Friedhelm Herrmann's laboratories. PMID- 11012250 TI - CML vaccines as a paradigm of the specific immunotherapy of cancer. AB - T cells are implicated in the effective control of chronic myeloid leukemia (CML). Recently, several clinical observations supported by laboratory data, indicate the presence of CML-specific T cells. Many proteins potentially act as leukemia-specific antigens for MHC-restricted cytotoxicity in CML. These include the bcr-abl fusion protein, myeloid-specific differentiation antigens and minor histocompatibility antigens. There is recent evidence to suggest that bcr-abl junctional peptides are capable of eliciting both CD4 and CD8 responses in normal healthy donors and in patients with CML. Moreover, T cell lines can be generated that react with autologous or HLA-matched fresh CML cells, suggesting that the bcr-abl fusion protein can be processed and expressed in the MHC cell surface molecules. Clinical trials exploiting the new understanding of the immunology of CML are underway. PMID- 11012251 TI - Gene regulation and deregulation: a beta globin perspective. AB - The study of the beta globin gene has provided great insights into the mechanisms of gene regulation and expression. In this review, we consider the normal regulation and expression of the beta globin gene and illustrate how the various steps may be affected, providing a basis for understanding the molecular pathophysiology of beta thalassemia. Mutations causing beta thalassemia can be classified as beta0 or B+ according to whether they abolish or reduce the production of beta globin chains. The vast majority of beta thalassemia is caused by point mutations, mostly single base substitutions, within the gene or its immediate flanking sequences. Rarely, beta thalassemia is caused by major deletions of the beta globin cluster. All these mutations behave as alleles of the beta locus but in several families the beta thalassemia phenotype segregates independently of the beta globin complex, and are likely to be caused by mutations in trans-acting regulatory factors. PMID- 11012252 TI - Reducing the risk of infection from plasma products: specific preventative strategies. AB - Collection and testing procedures of blood and plasma that are designed to exclude donations contaminated by viruses provide a solid foundation for the safety of all blood products. Plasma units may be collected from a selected donor population, contributing to the exclusion of individuals at risk of carrying infectious agents. Each blood/plasma unit is individually screened to exclude donations positive for a direct (e.g., viral antigen) or an indirect (e.g. anti viral antibodies) viral marker. As infectious donations, if collected from donors in the testing window period, can still be introduced into manufacturing plasma pools, the production of pooled plasma products requires a specific approach that integrates additional viral reduction procedures. Prior to the large-pool processing, samples of each donation for fractionation are pooled ('mini-pool') and subjected to a nucleic acid amplification test (NAT) by, for example, the polymerase chain reaction (PCR) to detect viral genomes (in Europe: HCV RNA plasma pool testing is now mandatory). Any individual donation found PCR positive is discarded before the industrial pooling. The pool of eligible plasma donations (which may be 2000 litres or more) may be subjected to additional viral screening tests, and then undergoes a series of processing and purification steps that, for each product, comprise one or several reduction treatments to exclude HIV, HBV HCV and other viruses. Viral inactivation treatments most commonly used are solvent-detergent incubation and heat treatment in liquid phase (pasteurization). Nanofiltration (viral elimination by filtration), as well as specific forms of dry-heat treatments, have gained interest as additional viral reduction steps coupled with established methods. Viral reduction steps have specific advantages and limits that should be carefully balanced with the risks of loss of protein activity and enhancement of epitope immunogenicity. Due to the combination of these overlapping strategies, viral transmission events of HIV, HBV, and HCV by plasma products have become very rare. Nevertheless, the vulnerability of the plasma supply to new infectious agents requires continuous vigilance so that rational and appropriate scientific countermeasures against emerging infectious risks can be implemented promptly. PMID- 11012253 TI - Cost effectiveness of sildenafil calls for political discussion. PMID- 11012254 TI - Debate about medical treatment of life prisoners. PMID- 11012255 TI - General practitioners have important roles in cancer. PMID- 11012256 TI - Reviving academic medicine in Britain. More honorary chairs are needed. PMID- 11012257 TI - Reviving academic medicine in Britain. Research and education must be given equal weight. PMID- 11012258 TI - Improving education for senior house officers. PMID- 11012259 TI - Preventive home visits to elderly people in the community. Visits are most useful for people aged >/= 75. PMID- 11012260 TI - Preventive home visits to elderly people in the community. Studies reviewed have methodical flaws. PMID- 11012261 TI - Preventive home visits to elderly people in the community. Further research is needed. PMID- 11012262 TI - Ethnicity and analgesia in accident departments. Authors did not exclude type II error or perform power calculation. PMID- 11012264 TI - Palliative Care Congress. Warwick, United Kingdom, 27-29 March 2000. Abstracts. PMID- 11012263 TI - Magnets and children--an attractive combination? PMID- 11012265 TI - [National Days of Laboratory Medicine of Russia. Moscow, October 12-14, 1999. Workshop: Laboratory Medicine on the Eve of the XXIst Century: synthesis of traditions and novel trends (analysis, diagnosis, technology, and economy). Abstracts]. PMID- 11012266 TI - New model law aims to expand criteria used by states for involuntary treatment of mental illness. PMID- 11012267 TI - Despite high compliance, GAO survey finds loopholes limit effectiveness of federal mental health parity law. PMID- 11012268 TI - Federal oversight of psychiatric hospitals needs to be strengthened, HHS watchdog concludes after review. PMID- 11012269 TI - Almost half of Americans report suicide attempt by close friend or relative. PMID- 11012270 TI - Chasing hyponatraemia in preterm infants. PMID- 11012271 TI - Low soluble FcRIII receptor demonstrates reduced neutrophil reserves in preterm neonates. PMID- 11012272 TI - Suxamethonium is safe in safe hands; mivacurium should also be considered. PMID- 11012273 TI - Incidence of severe retinopathy of prematurity. PMID- 11012274 TI - Premedication for intubation in neonates. PMID- 11012275 TI - Blood pressure standards for very low birthweight infants. PMID- 11012276 TI - Patient triggered ventilation in neonatal respiratory distress syndrome. PMID- 11012277 TI - Pyridoxine dependent epilepsy: a suggestive electroclinical pattern. PMID- 11012278 TI - Parental visiting in neonatal units. PMID- 11012279 TI - Ureaplasma colonisation and chronic lung disease in neonates. PMID- 11012280 TI - Stamps in neonatology. Incubators. PMID- 11012281 TI - [Complement--clinical significance of hypocomplementemia]. PMID- 11012282 TI - A specific electrotherapy technique in the treatment of osteoarthritis of the knee: three case reports. PMID- 11012283 TI - Qigong: a personal experience. PMID- 11012284 TI - [Is palliative non-fractionated radiotherapy really the same as palliative fractionated radiotherapy?] . PMID- 11012285 TI - Capture, binding, and detection technologies in clinical diagnostics. Proceedings and abstracts of the 32nd annual Oak Ridge Conference. 5-6 May 2000, Boston, Massachusetts, USA. PMID- 11012286 TI - When You're 24, 34 ... 84? PMID- 11012287 TI - World cardiovascular disease burden. PMID- 11012289 TI - 5th National Conference on Anticoagulant Therapy. May 14-15, 1999. Vancouver, British Columbia, Canada. Proceedings and abstracts. PMID- 11012288 TI - Myocardial infarction redefined. PMID- 11012290 TI - [Lipodystrophy syndrome. After 3 years every third patient has lost his fat]. PMID- 11012291 TI - [Nutritional therapy in HIV infection. Many approaches, meager data]. PMID- 11012292 TI - [Switch strategy: replacing the protease inhibitor. Quality of life improves]. PMID- 11012293 TI - [Ritonavir/Saqinavir: dual protease inhibitor combination. Salvage therapy]. PMID- 11012294 TI - Electrical stimulation of the renal nerve neither replicates its natural burst pattern nor proves the importance of that pattern for renal function. PMID- 11012295 TI - Changing strategies for treatment of systemic mycoses. AB - There have been a number of changes in strategies in antifungal therapy in the past few years. AIDS related mycoses have decreased, and the increase of fluconazole resistant Candida albicans may be slowing because fewer severely immune depressed patients require constant fluconazole suppression. Candida species continue to be relatively common blood culture isolates. About half of these are C. albicans and half non-albicans species. In recent years, we have moved from the use of amphotericin B to fluconazole for initial treatment of candidemia. We have seen fluconazole resistant isolates emerge, primarily C. glabrata and a few C. krusei, but also C. albicans. It is unclear whether the increasing use of fluconazole in intensive care units will worsen this problem. There appears to be no advantage for the lipid formulations of amphotericin B, though they are useful to reduce or prevent renal toxicity. In the United States and Europe, prevention and treatment of aspergillosis have become increasingly important. There are increasing data suggesting that lipid formulations are more effective for both treatment and prevention of invasive disease in the most vulnerable patients with this infection. Renal toxicity is reduced but not avoided by use of the lipid formulations of amphotericin B. For those patients with less acutely progressing disease, the triazoles may be effective options. It is unclear at present whether itraconazole, voriconazole, or posaconazole will be the most favored drug. One promising new class, now in clinical trials, is the echinocandin group. Other agents, such as the sordarins, the chitin synthase inhibitors, and topoisomerase inhibitors, have promise but are much earlier in development. Unfortunately, we still have >50% treatment failure with acute invasive aspergillosis, and 20%-30% failures with candidemia. Now that we have multiple classes of antifungal drugs available, and others in preclinical trials, it would be advantageous to begin more active exploration of combination therapy with antifungals and with combined immune modulators and antifungals. PMID- 11012296 TI - Antifungal susceptibility testing: progress and future developments. AB - The establishment of a standardized broth reference method for antifungal susceptibility testing of yeasts has opened the door to a number of interesting and useful developments. The adaptation of the reference macrodilution method to a microdilution method has significantly increased the clinical utility of antifungal susceptibility testing, and both methods are now included in the NCCLS document M27-A. The publication of quality control limits for five antifungal agents, coupled with the establishment of interpretive MIC breakpoints for three agents, provides useful parameters to survey clinical isolates of Candida and other yeast species. Adaptations of the M27 microdilution method for testing molds has also proved feasible. These developments have made it possible for a number of recent studies designed to expand the capabilities of laboratories to perform antifungal susceptibility testing and to enhance our understanding of trends in antifungal susceptibility. The availability of reference methods also provides a useful touchstone for the development of commercial products that promise to be more user friendly and to further improve test standardization. Products in varying stages of development include two colorimetric microdilution methods (Sensititre and KPI) and the Etest stable gradient MIC method. Preliminary data indicate that these methods are viable alternatives to the reference method for testing of yeasts. Furthermore, Etest may also prove useful for testing molds. Future expectations for antifungal susceptibility testing includes improved ability to detect amphotericin B resistance, development of an NCCLS document for susceptibility testing of molds, and application of these methods for testing dermatophytes. Incorporation of antifungal susceptibility testing methods into the clinical trials of new antifungal agents will facilitate the establishment of clinical correlates and further enhance the clinical utility of antifungal susceptibility testing. PMID- 11012297 TI - Atherosclerosis is a paradigmatic disease of the elderly, the roots of which are laid in youth, whereas the clinically manifested consequences become evident at old age. PMID- 11012298 TI - "New" risk factors for atherosclerotic diseases. AB - Many potential "new" risk factors may predispose to atherosclerotic diseases. The most promising currently seem to be fibrinogen, C-reactive protein and other inflammatory markers; hyperhomocysteinemia; low vitamin E intake; insulin resistance; lipoprotein(a); and fetal undernutrition. However, clinical trial proof of a causal role between several of these risk factors and CHD may be difficult to obtain unless selective interventions are developed to lower them. Certainly, current emphasis for primary and secondary prevention needs to focus on the established risk factors, particularly those for which clinical trial evidence has demonstrated interventions are efficacious for CHD prevention. PMID- 11012299 TI - Early diagnosis and treatment of pancreatic dysplasia in patients with a family history of pancreatic cancer. PMID- 11012300 TI - Prospective analysis of endoscopic papillary balloon dilatation and endoscopic sphincterotomy for removal of common bile duct stones. PMID- 11012301 TI - Clinical evaluation of endoscopic injection sclerotherapy using N-butyl-2 cyanoacrylate for gastric variceal bleeding. PMID- 11012302 TI - Gamma-hydroxy butyrate withdrawal delirium: a case report. PMID- 11012303 TI - Self-inflicted bilateral eye injury by a schizophrenic patient. PMID- 11012304 TI - Beyond hemiplegia. PMID- 11012305 TI - The 2nd S. Takahashi Memorial International Workshop on 3-Dimensional Conformal Radiotherapy. Nagoya, Japan, December 11-13, 1998. PMID- 11012306 TI - Regarding scoring of radiation pneumonitis. PMID- 11012307 TI - Defining a uniform biologically effective dose for organs with parallel architecture. PMID- 11012308 TI - Fraction size and dose parameters related to the incidence of pericardial effusions: regarding Martel et al. IJROBP 40(1):155-161; 1998. PMID- 11012309 TI - Optimizing for the Gamma knife. PMID- 11012310 TI - Response to Drs. Bauman et al.: pretreatment factors predict overall survival for patients with low-grade glioma: a recursive partitioning analysis (IJROBP 1997;45[4]:923-929, 1999) PMID- 11012311 TI - Should the delta-TCP concept be replaced by "local control probability ratios"? PMID- 11012312 TI - Role of magnesium in alleviating pain: newer insights. PMID- 11012313 TI - Painful gynecomastia: an unusual toxicity of gabapentin? PMID- 11012314 TI - Visualization of the vesicular acetylcholine transporter in living cholinergic cells. PMID- 11012315 TI - [Spring reunion of the Southwest Delegation of the SFR (SFRIMSO)--French Society of Radiology and Medical Imaging of the Southwest Region]. PMID- 11012316 TI - [Coordination meeting on the prevention of transfrontier transmission of malaria in selected countries of the WHO East Mediterranean and European Regions]. PMID- 11012317 TI - [The historical experience of malaria control (on the 75th anniversary of the resolution of the Soviet People's Commissariat of the RSFSR, Measures for the Control of Malaria)]. PMID- 11012318 TI - [The current problems of malariology (a survey of the materials from the international conference on malaria, the MIM African Malaria Conference, Durban, South Africa, 14-19 March 1999). Multilateral Initiative on Malaria]. PMID- 11012319 TI - Sleep-disordered breathing and hypertension. PMID- 11012320 TI - Sleep-disordered breathing and hypertension. PMID- 11012321 TI - Sleep-disordered breathing and hypertension. PMID- 11012322 TI - Hemodynamic effects of sildenafil. PMID- 11012323 TI - Use of right precordial leads during exercise testing. PMID- 11012324 TI - Haptoglobin phenotype and vascular complications in patients with diabetes. PMID- 11012325 TI - Fortification of foods with folic acid. PMID- 11012326 TI - Fortification of foods with folic acid. PMID- 11012327 TI - Fortification of foods with folic acid. PMID- 11012328 TI - Fortification of foods with folic acid. PMID- 11012329 TI - Life-threatening toxic epidermal necrolysis with thalidomide therapy for myeloma. PMID- 11012330 TI - Improvement of Huntington's disease with olanzapine and valproate. PMID- 11012331 TI - Signal transduction. An arresting tale. PMID- 11012332 TI - Resuscitation of a choking child in 1841. PMID- 11012333 TI - Timing and etiology of neonatal cerebral infarction. PMID- 11012334 TI - Universal neonatal hearing screening. PMID- 11012336 TI - Cardiopulmonary resuscitation in very low birth weight infants. PMID- 11012335 TI - Universal neonatal hearing screening. PMID- 11012337 TI - First urinary tract infections in Swedish children. PMID- 11012338 TI - Contraception and adolescents. PMID- 11012339 TI - Current age of onset of puberty. PMID- 11012340 TI - On looking inward and being scientific: a tribute to G.L. Engle, MD. PMID- 11012341 TI - Chemoradiation for carcinoma of the cervix: advances and opportunities. AB - Although it is possible to cure many patients with locally advanced cervical cancer using radiation therapy alone, loco-regional relapse continues to be a component of most recurrences. To improve control rates, clinicians have investigated ways of combining chemotherapy and radiation for more than 30 years. Despite encouraging results from phase II trials of neoadjuvant chemotherapy, randomized trials failed to improve on the results with radiation therapy alone. For a number of reasons, early trials of concurrent chemoradiation were inconclusive. However, recent reports of five large prospective randomized trials demonstrated dramatic improvements in survival and local control rates when cisplatin-containing chemotherapy was given during radiation therapy. These results also suggest a number of avenues for future research. PMID- 11012342 TI - Identification of potential mRNA biomarkers in peripheral blood lymphocytes for human exposure to ionizing radiation. AB - Since early in the Atomic Age, biological indicators of radiation exposure have been sought, but currently available methods are not entirely satisfactory. Using cDNA microarray hybridization to discover new potential biomarkers, we have identified genes expressed at increased levels in human peripheral blood lymphocytes after ex vivo irradiation. We recently used this technique to identify a large set of ionizing radiation-responsive genes in a human cell line (Oncogene 18, 3666-3672, 1999). The present set of radiation markers in peripheral blood lymphocytes was identified 24 h after treatment, and while the magnitude of mRNA induction generally decreased over time, many markers were still significantly elevated up to 72 h after irradiation. In all donors, the most highly responsive gene identified was DDB2, which codes for the p48 subunit of XPE, a protein known to play a crucial role in repair of ultraviolet (UV) radiation damage in DNA. Induction of DDB2, CDKN1A (also known at C1P1/WAF1) and XPC showed a linear dose-response relationship between 0.2 and 2 Gy at 24 and 48 h after irradiation, with less linearity at earlier or later times. These results suggest that relative levels of gene expressions in peripheral blood cells may provide estimated of environmental radiation exposures. PMID- 11012344 TI - Mutations induced by alpha-particle radiation--no evidence for a bystander effect. PMID- 11012343 TI - Proficient repair of potentially lethal damage sensitive to hypertonic treatment in osteosarcoma cells. AB - Fast-repairing potentially lethal damage (PLD) in seven osteosarcoma cell lines was analyzed after treatment with a hypertonic 0.5 M NaCl solution for 20 min and compared to that in seven human fibroblast strains. Fixation of PLD after exposure to ionizing radiation was observed without exception in both the osteosarcoma cells and the fibroblast strains. The percentages by which the D(o)'s of the osteosarcoma cells decreased were significantly higher than the percentage decreases in the C(o)'s of the fibroblast strains (P < 0.01). Hypertonic treatment resulted in radiosensitization due to fixation of PLD in all of the osteosarcoma cell lines, demonstrating that osteosarcoma cells can repair PLD better than normal fibroblast cells. The radiobiological response of the osteosarcoma cells, with enhanced killing after hypertonic treatment, was similar to that of normal untreated fibroblast cells. PMID- 11012345 TI - [Optimizing radiation protection of patients and nurses]. PMID- 11012346 TI - Forensic epidemiology. Vaccine theory of AIDS origins disputed at Royal Society. AB - At center stage at a meeting on the origin of the AIDS epidemic, held here this week at the Royal Society, was a controversial theory that a contaminated polio vaccine tested in Africa more than 40 years ago sparked the epidemic. The theory took a hit when researchers revealed that tests of old samples of the vaccine provided no supporting evidence, and the main proponent of the theory, British writer Edward Hooper, endured a verbal battering himself from several prominent scientists. But Hooper, unbowed, got in plenty of jabs of his own. PMID- 11012347 TI - Biomedicine. 'Glue grant' boosts cell signaling consortium. PMID- 11012348 TI - U.S. postdocs. Report urges better treatment, status. PMID- 11012349 TI - Appointments. Salk Institute goes north for new CEO. AB - The revolving door at the top of the Salk Institute for Biological Studies in La Jolla, California, took another spin last week with the appointment of neuroscientist Richard Murphy as president and CEO. Murphy, director of the Montreal Neurological Institute, will become Salk's fourth chief executive in 4 years when he takes up the reins on 1 October. Murphy, 56, says his main job will be to raise enough money to keep the endowment-poor research institute in the scientific big leagues. PMID- 11012350 TI - Molecular biology. Cancer fighter's modus operandi revealed. AB - Researchers have deciphered how a promising cancer drug acts like a smart bomb, homing in on only a very narrow range of its potential targets in the cell. The compound, known as STI-571, has shown remarkable success in early clinical trials on patients with chronic myelogenous leukemia. Now, in work reported on page 1938, scientists reveal just how the compound works--information that could aid in the design of similar cancer therapies. PMID- 11012351 TI - European science. Call to arms for life scientists. PMID- 11012352 TI - Virology. Evolution on life's fringes. AB - Earlier this summer, two dozen scientists gathered here to take another crack at the question of viral origins and evolution. Fresh evidence that viruses have existed for billions of years suggests that they were on hand when the first cells arose. That has scientists wondering what role these stripped-down microbes played in the evolution of life. PMID- 11012353 TI - Biochemistry. High-tech lures hook into new marine microbes. AB - Two scientific teams, one of which reports its findings on page 1902, have identified large populations of bacteria that convert sunlight hitting the sea surface into energy. These two groups of bacteria, which harness light to move electrons and power cellular processes, likely help bring energy into the food chain. And that could help to explain a puzzle that has confounded researchers trying to understand marine ecosystems: how so many bacteria can survive in the open ocean, where there seems to be relatively little for them to eat. PMID- 11012354 TI - Conservation in the real world. PMID- 11012355 TI - The NIH guidelines on stem cell research. PMID- 11012356 TI - How to resolve the debate on the origin of AIDS. PMID- 11012357 TI - An early taste of things to come. PMID- 11012358 TI - Not-so-simple minds. PMID- 11012359 TI - PCBs are a health risk for humans and wildlife. PMID- 11012360 TI - Intellectual property. Publication rights in the era of open data release policies. AB - The open data release policy adopted by the large-scale DNA sequencing centers has made accessible valuable information that facilitates research. Herein, we argue that the data producers' rights to receive credit for at least some portion of the analyses of the data must be protected. We suggest that this protection take the form of a specification of the probable content of the primary paper the data producers intend to publish when the data gathering is complete. Rights to publish that paper ought then be restricted to the producers unless they give permission otherwise. PMID- 11012361 TI - Evolution. Terrestrial life--fungal from the start? AB - Fungi have been implicated in the early colonization of land by plants. As Blackwell explains in her Perspective, Redecker et al.'s discovery of a fossil fungus provides strong support for this hypothesis. The important role of symbiotic associations of fungi with plants and animals is increasingly being recognized, as is the fact that some of these associations date back to the origin of terrestrial life on Earth. PMID- 11012362 TI - Cloning. Pigs is pigs. AB - Since the first report of a cloned animal (Dolly the sheep) 3 years ago, cloning mammals has become something of a cottage industry. As Prather discusses in his Perspective, pigs can now be added to the august list of cloned animals, which includes cows, goats, and mice. This is a particularly spectacular achievement because pig cloning has turned out to be notoriously difficult. The pig is also a valuable domestic animal to have cloned because, being physiologically close to humans, its organs can be used in xenotransplantation. PMID- 11012363 TI - Neuroscience. Regional differences in cortical organization. AB - Although there are elegants maps of the human brain that reveal differences in cellular architecture between different cortical regions, there is not much information about how corresponding cortical regions differ between the left and right hemispheres. As Gazzaniga explains in his Perspective, new results reveal the surprising finding of asymmetry in area 22 (which is important for language processing) of the left and right hemisphere (Galuske et al.). Clusters of neurons in area 22 of the left hemisphere are spaced farther apart and have longer axons cabling them together than neuronal clusters in area 22 of the right hemisphere. PMID- 11012364 TI - Tech.sight. Genetic testing--present and future. PMID- 11012365 TI - [Correct usage of drugs. 8/12--Preparation and administration of injectable drugs]. PMID- 11012366 TI - [Nursing responsibility. 9/13--The responsibility of the pediatric nurse]. PMID- 11012367 TI - [Management of an imminent home delivery]. PMID- 11012368 TI - [Hygienic care and prevention. 3--shampooing]. PMID- 11012369 TI - PEF versus FEV1. PMID- 11012370 TI - Caring for indigenous Australian children with asthma. PMID- 11012371 TI - Differentiation between mitral stenosis and coexisting PPH. PMID- 11012372 TI - Intravenous montelukast in acute asthma: expensive aminophylline? PMID- 11012374 TI - Initial platelet activity may predict efficacy after chronic oral glycoprotein IIb/IIIa blockade: should we still consider uniform treatment regimens? PMID- 11012373 TI - TB guidelines. PMID- 11012375 TI - Does clinical significance exist in small platelet aggregates detected by the laser light scattering method? PMID- 11012376 TI - The effects of hormone replacement therapy on thrombin generation, fibrinolysis inhibition, and resistance to activated protein C: prospective cohort study and review of literature. AB - Recent studies have found that hormone replacement therapy (HRT) is associated with a two- to fourfold increased risk of venous thromboembolism, but the thrombogenic mechanism of HRT remains unclear. To investigate whether HRT use induces a procoagulant state, we undertook a prospective cohort study in postmenopausal women to investigate the effects of 3 months of treatment with oral HRT (conjugated equine estrogen 0.625 mg daily and medroxyprogesterone 2.5 mg daily) on markers of thrombin generation (prothrombin fragment 1+2, thrombin antithrombin complexes), fibrinolytic potential (plasminogen activator inhibitor 1 (PAI-1) activity), and activated protein C (APC) resistance. In addition, we reviewed the literature for studies investigating the effects of HRT on markers of thrombin generation and fibrinolytic potential. In 12 patients who received HRT for a mean of 3.8 months, there was no significant effect of HRT on levels of F1+2, thrombin-antithrombin complexes, or the APC ratio. HRT use had the greatest effect on PAI-1 activity (mean difference = -3.75 UI/mL; 95% confidence interval: - 8.9, 1.1) compared to other coagulation parameters, but this did not attain statistical significance (p = 0.12). In the literature review, the effects of HRT on markers of thrombin generation were inconsistent across studies. There was a consistent pattern of increased fibrinolytic potential with HRT use associated with one marker (PAI-1), but not with another marker (tissue plasminogen activator antigen). We conclude that there is a lack of consistent evidence that the increased risk of venous thromboembolism associated with HRT use is due to a procoagulant state related to increased thrombin generation, decreased fibrinolytic potential, or acquired APC resistance. PMID- 11012378 TI - [Academician Boris Alekseevich Korolev (on his 90th birthday)]. PMID- 11012377 TI - In vitro characterization of a novel factor Xa inhibitor, RPR 130737. AB - RPR 130737 inhibited factor Xa (FXa) with a Ki of 2.4 nM and also displayed excellent specificity toward FXa relative to other serine proteases. It showed selectivity of more than 1000-fold over thrombin, activated protein C, plasmin, tissue-plasminogen activator and trypsin. RPR 130737 prolonged plasma activated partial thromboplastin time and prothrombin time in a dose-dependent fashion. In the activated partial thromboplastin time assay, the concentrations required for doubling coagulation time were 0.32 microM (human), 0.61 microM (monkey), 0.44 microM (dog), 0.15 microM (rabbit), and 0.82 microM (rat). The concentrations required to double prothrombin time were 0.86 microM (human), and 1.26 microM (monkey), 1.15 microM (dog), 0.39 microM (rabbit) and 7.31 microM (rat). Kinetic studies revealed that RPR 130737 was a fast binding, reversible and competitive inhibitor for FXa when Spectrozyme FXa, a chromogenic substrate, was used. A coupled-enzyme assay measuring thrombin activity following prothrombinase conversion of prothrombin to thrombin indicated that RPR 130737 was a potent inhibitor for prothrombinase-bound FXa. In this assay, RPR 130737 showed IC50s of 17 nM and 35.9 nM, respectively when artificial phosphatidylserine/phosphatidylcholine (PS/PC) liposomes or gel-filtered platelets were used as the phospholipid source. An FX-deficient plasma clotting time correction assay further demonstrated that RPR 130737 was a specific inhibitor of FXa. RPR 130737 showed no effect on platelet aggregation in vitro. These results indicate that RPR 130737 has the potential to be developed as an antithrombotic agent based on its potent and selective inhibitory effect against FXa. PMID- 11012379 TI - [Cardiac Imaging 2000. Proceedings of a symposium. 4-5 June 1999]. PMID- 11012380 TI - [Fifty years of the Institute of Higher Nervous Activity and Neurophysiology of the Russian Academy of Sciences]. PMID- 11012381 TI - [42nd Kasseler Symposium on Surgical Diseases of the Bowel. 25-26 June 1999, Baunatal. Abstracts]. PMID- 11012382 TI - [New vaccines developed in the Russian Federation]. PMID- 11012384 TI - 50th Anniversary Meeting of the Bone and Tooth Society. 10-12 July 2000, Cambridge, United Kingdom. Abstracts. PMID- 11012383 TI - [The registration and postregistration control of foreign vaccines]. PMID- 11012385 TI - Possible mechanisms of increased blood viscosity in systemic hypertension. PMID- 11012386 TI - Arrhythmic right ventricular cardiomyopathy. PMID- 11012387 TI - A view on the science: physical anthropology at the millennium. PMID- 11012388 TI - Biomechanics of Mammalian Feeding and Primate Evolution. Proceedings of a symposium. Research Triangle Park, North Carolina, USA. April 9-13, 1996. PMID- 11012389 TI - Molecular analysis of Tn1546 in vanA-containing Enterococcus spp. isolated from humans and poultry. PMID- 11012390 TI - Establishing criteria for assessment of efficacy of antimicrobial agents in acute otitis media. PMID- 11012392 TI - Proceedings of the 21st Symposium on Biotechnology for Fuels and Chemicals. May 2 6, 1999. Fort Collins, Colorado, USA. PMID- 11012391 TI - Efavirenz-induced decrease in plasma amprenavir levels in human immunodeficiency virus-infected patients and correction by ritonavir. PMID- 11012393 TI - Aged dialyzers and vision loss. PMID- 11012394 TI - Underscoring the importance of mentoring. PMID- 11012395 TI - Onderstepoort distemper vaccine. PMID- 11012396 TI - Index of suspicion. Case 3. Acetaminophen overdose. PMID- 11012397 TI - Stakeholder analysis: a review. AB - The growing popularity of stakeholder analysis reflects an increasing recognition of how the characteristics of stakeholders--individuals, groups and organizations -influence decision-making processes. This paper reviews the origins and uses of stakeholder analysis, as described in the policy, health care management and development literature. Its roots are in the political and policy sciences, and in management theory where it has evolved into a systematic tool with clearly defined steps and applications for scanning the current and future organizational environment. Stakeholder analysis can be used to generate knowledge about the relevant actors so as to understand their behaviour, intentions, interrelations, agendas, interests, and the influence or resources they have brought--or could bring--to bear on decision-making processes. This information can then be used to develop strategies for managing these stakeholders, to facilitate the implementation of specific decisions or organizational objectives, or to understand the policy context and assess the feasibility of future policy directions. Policy development is a complex process which frequently takes place in an unstable and rapidly changing context, subject to unpredictable internal and external factors. As a cross-sectional view of an evolving picture, the utility of stakeholder analysis for predicting and managing the future is time limited and it should be complemented by other policy analysis approaches. PMID- 11012398 TI - How to act--implementing health and safety promotion in organizations. AB - This interdisciplinary review focuses on strategies for implementing health and safety promotion activities in organizations. Theories are summarized in a checklist and illustrated by some practical examples from Sweden and abroad. The points illustrated appear obvious and logical, but they are seldom applied in practise. PMID- 11012399 TI - A planning framework for community empowerment goals within health promotion. AB - Health promotion often comprises a tension between 'bottom-up' and 'top-down' programming. The former, more associated with concepts of community empowerment, begins on issues of concern to particular groups or individuals, and regards some improvement in their overall power or capacity as the important health outcome. The latter, more associated with disease prevention efforts, begins by seeking to involve particular groups or individuals in issues and activities largely defined by health agencies, and regards improvement in particular behaviours as the important health outcome. Community empowerment is viewed more instrumentally as a means to the end of health behaviour change. The tension between these two approaches is not unresolvable, but this requires a different orientation on the part of those responsible for planning more conventional, top-down programmes. This article presents a framework intended to assist planners, implementers and evaluators to systematically consider community empowerment goals within top-down health promotion programming. The framework 'unpacks' the tensions in health promotion at each stage of the more conventional, top-down programme cycle, by presenting a parallel 'empowerment' track. The framework also presents a new technology for the assessment and strategic planning of nine identified 'domains' that represent the organizational influences on the process of community empowerment. Future papers analyze the design of this assessment and planning methodology, and discuss the findings of its field-testing in rural communities in Fiji. PMID- 11012400 TI - Health policy-making in central and eastern Europe: lessons from the inaction on injuries? AB - The burden of disease due to injuries has elicited virtually no public health response in the countries of central and eastern Europe, even though injuries have long been a much greater problem in the east of Europe than in the west, with children especially affected. This paper seeks to identify factors that have inhibited policy development on this topic and to draw lessons for health policy development in this region more generally. Several factors emerge. Deaths from injuries have had low visibility. Data have not been assembled in a way that would facilitate identification of the burden of disease that they constitute. Those organizations responsible for public health, whether within government or at local level, were typically very weak with little capacity either to identify the nature and scale of threats to the health of their populations or to develop strategies to address them. There was uncertainty about ownership, with fragmentation of responsibility but no tradition of intersectoral working. Non governmental organizations, which have placed injuries on the health policy agenda in the west, are weak or non-existent. International donors, who could have had a role, have focused on issues such as health care reform. This analysis provides a potential framework for examining policy responses, or lack thereof, to other health challenges in this region. It highlights the need for a better understanding of the potential for using available data, which, in turn, requires a major strengthening of capacity. However, in many countries, there is a need for new ways of working, involving a broadening of the sense of ownership, with clearly designated responsibilities but designed in ways that encourage rather than inhibit intersectoral action. There is also a need to develop non governmental organizations that have sufficient capacity to undertake their own analyses and to place issues on the agenda. PMID- 11012401 TI - Hospital practices in maternity wards in Lebanon. AB - This study was conducted in Lebanon with the main objectives of acquiring baseline data on practices and routines applied in the obstetrics ward for women having normal delivery; estimating the frequency of certain practices; and assessing whether women are given choice in these practices. A national sample of 39 hospitals was selected. The director, head midwife, or head nurse of the obstetrics department was interviewed using a semi-structured questionnaire. The hospitals studied are largely equipped to cope with emergencies and services are available 24 hours a day. On average, the caesarean section rate is 18% and the stillbirth rate is 10 per 1000, but with considerable variability between facilities. The majority of hospitals do not have written policies or standard birth procedures and lack mechanisms for evaluation. Generally, minimal prenatal information is given to women. Companions are allowed during labour but this access is more restricted in delivery. The reported configuration of professional care during labour and delivery is favourable to high quality care. In terms of mobility during labour, most hospitals allow women to move. However, 31 hospitals set an i.v. drip to all women and some use continuous fetal monitoring method. Mobility is restricted in delivery; in 23 hospitals women are tied down. Nearly all hospitals give intra-muscular anaesthesia whereas epidurals are used less frequently. As for postpartum care, most hospitals do not initiate breastfeeding within one hour of birth and few have rooming in. The majority of hospitals do not provide women with family planning methods and a few do not even discuss methods with them. The approach used in this study constitutes a tool for understanding and assessing maternity services that should be applied in other settings. The tool is available from the authors. PMID- 11012402 TI - Improving the monitoring of immunization services in Kyrgyzstan. AB - Following the disbanding of the Soviet Union in 1991, the government of Kyrgyzstan was unable to maintain the previous level of health services. To revitalize the health services, the Ministry of Health (MOH) first focused on improving their immunization services, including the immunization component of the Health Management Information System (HMIS). Secondly, to increase immunization coverage, the MOH set as a priority the elimination of prescribing false contraindications to immunization. To accomplish both goals, the MOH updated the national immunization policies and established a more effective structure for managing immunization services. To support the MOH, the US Agency for International Development (USAID) Resources for Child Health (REACH) and Basic Support for Institutionalizing Child Survival (BASICS) projects provided technical assistance through a resident coordinator and consultants, and by organizing an international seminar. The improvements extended beyond systems and forms, but, instead, emphasized monitoring by the frontline health worker and supervising the quality of health information. To accomplish their objectives, the MOH appointed a Working Group to define the problems, revise record-keeping procedures, and develop monitoring tools. This group, representing both national and local levels, was composed of MOH epidemiologists, paediatricians and a management information specialist. To reduce the burden of excessive record keeping and reporting requirements, the Working Group identified four key indicators for the service delivery level: (1) DPT3 immunization coverage rates for children less than 1 year of age; (2) contraindication rates for DPT; (3) usage of DPT vaccine; and (4) daily refrigerator temperatures. Additional indicators were included at district and provincial levels. After a successful 1 year trial, the MOH implemented the revised HMIS nationally. Not only did the quality of the information system improve, but the new approach provided visible evidence, from facility to national levels, that the MOH was approaching their objective of reducing contraindication rates for DPT immunizations to 5% or less, and that vaccine wastage could be substantially reduced. The project demonstrated that giving health workers the basic epidemiologic skills to monitor their own work measurably improved the quality of the data, and by acquiring the new skills, the workers developed a sense of pride in their work. PMID- 11012403 TI - Financing blood transfusion services in sub-Saharan Africa: a role for user fees? AB - The provision of a secure and safe blood supply has taken on new importance in sub-Saharan Africa with the onset of the AIDS epidemic. Blood transfusion services capable of providing safe blood are not cheap, however, and there has been some debate on the desirability and sustainability of different financing mechanisms for blood transfusion services. This paper examines patterns of financing blood transfusion in three countries--Cote d'Ivoire, Zimbabwe and Mozambique. It goes on to consider the conceptual options for financing safe blood, and to examine in detail the possible role of user fees for blood transfusion in Africa, developing a simple model of their likely burden to patients based on data from Cote d'Ivoire. The model indicates that, at best, there can only be a limited role for user fees in the financing of safe blood transfusion services, due mainly to the relatively high cost of producing a unit of safe blood. Charging individuals for the blood they receive is likely to be administratively complex and costly, could realistically recover only a fraction of the production costs involved, and is further complicated by the fact that the main recipients of blood transfusion in sub-Saharan Africa are children and pregnant women. If cost-recovery for safe blood is to be attempted, the most viable option appears to be that of charging a collective fee, levied upon all inpatients, not just on those who receive blood. Such a mechanism is not without problems, not least in its failure to offer incentives for more appropriate blood use, and it is still likely to recover only a portion of the costs of producing safe blood. Whether or not cost-recovery is instituted, there will remain an important role for public funding of blood transfusion services, and, by implication, an important role for foreign donor support. PMID- 11012404 TI - The paradox of the cost and affordability of traditional and government health services in Tanzania. AB - Since the introduction of user fee systems in the government health facilities of most African countries, which shifted part of the burden of financing health care onto the community, affordability of basic health care has been a much discussed topic. It is sometimes assumed that in areas where high levels of spending for traditional treatments are common, people would be able to pay for basic health care at governmental facilities, but may not be willing to do so. However, examining willingness to pay and ability to pay in the broader context of different types of illness and their treatment leads us to a very different conclusion. In the course of a medical-ethnographic study in south-eastern Tanzania, we found evidence that people may indeed be willing, but may nevertheless not be able, to pay for biomedical health care--even when they can afford costly traditional medicine. In this article, we suggest that the ability to pay for traditional treatment can differ from ability to pay for hospital attendance for two main reasons. First, many healers--in contrast to the hospital -offer alternatives to cash payments, such as compensation in kind or in work, or payment on a credit basis. Secondly, and more importantly, the activation of social networks for financial help is different for the two sectors. For the poor in particular, ability to pay for health care depends a great deal on contributions from relatives, neighbours and friends. The treatment of the 'personalistic' type of illness, which is carried out by a traditional healer, involves an extended kin-group, and there is high social pressure to comply with the requirements of the family elders, which may include providing financial support. In contrast, the costs for the treatment of 'normal' illnesses at the hospital are usually covered by the patient him/herself, or a small circle of relatives and friends. PMID- 11012405 TI - Financing reforms for the Thai health card scheme. AB - The Thai health card scheme originated from a pilot study on community financing and primary health care in maternal and child health in 1983. The scheme later changed to one of voluntary health insurance and finally received a matching subsidy from the government. The coverage of the scheme is described by a U curve, i.e. it started with 5% of the total population in 1987, declined to 3% in 1992, with an upturn to 14% in 1997. The upturn has been the result of concerns about universal coverage policy, together with reforms of fund management. The provincial fund is responsible for basic health, basic medical, referral, and accident and emergency services. The central fund takes 2.5% of the total fund to manage cross-boundary services and high cost care (a reinsurance policy). On average, the utilization rate of the voluntary health card was higher than that of the compulsory (social security) scheme. And amongst three variants of health cards, the voluntary health card holders used health services twice to three times more than the community and health volunteer card holders. Cost recovery was low, especially in the provinces with low coverage. In the province with highest coverage, cost recovery was as high as 90% of the non-labour recurrent cost. Only 10% of the budgeted fund for reinsurance was disbursed, implying considerable management inefficiency. The management information system as well as the management capacity of the Health Insurance Office should be strengthened. After comparing the health card with other insurance schemes in terms of coverage, cost recovery, utilization and management cost, it is recommended that this voluntary health insurance should be modified to be a compulsory insurance, with some other means of premium collection and minimal co-payment at the point of delivery. PMID- 11012406 TI - From state to market: the Nicaraguan labour market for health personnel. AB - Few countries in Latin America have experienced in such a short period the shift from a socialist government and centrally planned economy to a liberal market economy as Nicaragua. The impact of such a change in the health field has been supported by the quest for reform of the health system and the involvement of external financial agencies aimed at leading the process. However, this change has not been reflected in the planning of human resources for health. Trends in education reflect the policies of past decades. The Ministry of Health is the main employer of health personnel in the country, but in recent years its capacity to recruit new personnel has diminished. Currently, various categories of health personnel are looking for new opportunities in a changing labour environment where new actors are appearing and claiming an influential role. It may take more than political willingness from the government to redefine the new priorities in the field of human resources for health and subsequently turn it into positive action. PMID- 11012407 TI - Health sector reform and human resources: lessons from the United Kingdom. AB - The objective of the paper is to assess the human resource (HR) dimension of the National Health Service (NHS) reforms in the United Kingdom, and to highlight lessons for the health systems of countries undergoing reform or restructuring. Health sector reform in many countries in the 1980s and 1990s has focused on structural change, cost containment, the introduction of market mechanisms and consumer choice. This focus has inevitably challenged the ways that health professionals and other staff are employed and deployed. The methods used to manage human resources in health care may also in themselves be a major constraint or facilitator in achieving the objectives of health sector reform. The impact on the HR function of the NHS reforms is assessed in the paper by examining three central requirements of the HR function: to maintain effective staffing levels and skill mix; to establish appropriate employee relations policy and procedures; and to be involved in pay determination. The paper concludes that the most significant changes which have occurred as result of the NHS reforms have been in staffing change and organizational culture, and the individual attitudes of NHS management and staff. Attempts to alter methods of conducting employee relations and determining pay and conditions of employment have been less successful. However, an overall approach to HR management, which would have been unthinkable in the pre-reform NHS, is now accepted, albeit grudgingly by some, as the way forward. In general, the changes in the NHS HR function can be characterized as a partially successful attempt to adopt private sector HR management techniques to meet the challenges of public sector reform. PMID- 11012408 TI - The impact of traditional birth attendant training on delivery complications in Ghana. AB - OBJECTIVES: In their efforts to reduce maternal and neonatal morbidity and mortality, many national and international agencies make considerable investments in training traditional birth attendants (TBAs). The value of TBA training is controversial, and plausible arguments are made both for and against. Numerous process evaluations are reported in the literature and the results are mixed, though generally positive. Outcome evaluations, however, are scarce. This article describes an outcome evaluation of TBA training conducted in two districts of Brong-Ahafo Region, Ghana, during 1996. DESIGN AND METHODS: Data from a random sample survey of 1961 clients of TBAs were subjected to logistic regression modelling to determine the effect of training on maternal outcomes, controlling for other independent variables. RESULTS: Of eight outcomes modelled, three were associated with training and five were not. Three additional outcomes were not modelled, primarily due to low prevalence. CONCLUSIONS: Despite some inherent design limitations, this study found that the evidence for a beneficial impact of TBA training was not compelling. Training sponsors should consider alternative health investments and, where TBA training remains the intervention of choice, be realistic about expectations of impact. PMID- 11012409 TI - A participatory approach to sanitation: experience of Bangladeshi NGOs. AB - This study assesses the role of participatory development programmes in improving sanitation in rural Bangladesh. Data for this study came from a health surveillance system of BRAC covering 70 villages in 10 regions of the country. In depth interviews were conducted with one adult member of a total of 1556 randomly selected households that provided basic socioeconomic information on the households and their involvement with NGO-led development programmes in the community. The findings reveal that households involved with credit programmes were more likely to use safe latrines than others who were equally poor but not involved in such programmes. The study indicates that an unmet need to build or buy safe and hygienic latrines existed among those who did not own one. Such latent need could be raised further if health education at the grassroots level along with supervised credit supports were provided to them. Unlike conventional belief, the concept of community-managed jointly owned latrines did not seem a very attractive alternative. The study argues that social and behavioural aspects of the participatory development programmes can significantly improve environmental sanitation in a traditional community. PMID- 11012411 TI - Conferences. AB - International Conference on Humanization of Childbirth * November 2-4, 2000 * Fortaleza, Ceara, Brazil The XVth International Conference on the Social Sciences & Medicine - Societies & Health in Transition * October 16-20, 2000 * Eindhoven, The Netherlands PMID- 11012410 TI - A stakeholder analysis. AB - This paper provides guidance on how to do a stakeholder analysis, whether the aim is to conduct a policy analysis, predict policy development, implement a specific policy or project, or obtain an organizational advantage in one's dealings with other stakeholders. Using lessons learned from an analysis of alcohol policy development in Hungary, it outlines issues to be considered before undertaking the stakeholder analysis concerning the purpose and time dimensions of interest, the time-frame and the context in which the analysis will be conducted. It outlines advantages and disadvantages of an individual or team approach, and of the use of insiders and outsiders for the analysis. It describes how to identify and approach stakeholders and considers the use of qualitative or quantitative data collection methods for estimating stakeholder positions, levels of interest and influence around an issue. A key message is that the process of data collection and analysis needs to be iterative; the analyst needs to revise and deepen earlier levels of the analysis, as new data are obtained. Different examples of ways of analyzing, presenting and illustrating the information are provided. Stakeholder analysis is a useful tool for managing stakeholders and identifying opportunities to mobilize their support for a particular goal. However, various biases and uncertainties necessitate a cautious approach in using it and applying its results. PMID- 11012412 TI - Publications. AB - The World Health Organization (WHO) has recently published a booklet of Guidelines for the Regulatory Assessment of Medicinal Products for use in Self Medication. The guidelines' purpose is to suggest criteria and methods which drug regulatory authorities can use in determining the suitability of medicinal products for self-medication. PMID- 11012414 TI - Radiology 2000: david G. Disler completes his service as deputy editor PMID- 11012413 TI - Radiology 2000: ann S. Fulcher, MD, appointed senior deputy editor PMID- 11012415 TI - Congratulations to the 2000 editorial fellows PMID- 11012416 TI - Cardiac imaging. AB - The emergence of noninvasive imaging techniques for the definitive diagnosis and monitoring of cardiovascular disease has greatly altered cardiac imaging in the past 25 years. The practice of cardiac imaging in 1975 was centered on conventional radiography and angiography, but, in the past 2 decades, noninvasive techniques have substantially replaced catheterization and angiography. The reliance on echocardiography for the evaluation of many cardiac diseases had a profoundly negative influence on the role of the radiologist in cardiac imaging, since the exercise of this modality has been a nearly exclusive province of the cardiologist. However, in the past decade, magnetic resonance imaging has been gradually assuming more importance in cardiovascular diagnosis; with this increase in importance, the role of the radiologist has been reactivated. In 1975, fellowship training in cardiac imaging was frequently combined with training in angiography. Now, training may be more effective by combining cardiac and pulmonary imaging in a thoracic imaging fellowship, but cross-training with an associated subspeciality will be influenced by priorities and personnel in various departments. PMID- 11012417 TI - Managed care and social justice. PMID- 11012418 TI - Dynamic contrast-enhanced MR imaging of the scrotum: reality check. PMID- 11012419 TI - Posttransplantation lymphoproliferative disorder in children: clinical, histopathologic, and imaging features. AB - Posttransplantation lymphoproliferative disorder (PTLD) is a condition in patients who receive transplants in which chronic immunosuppression leads to an unregulated expansion of lymphoid cells; the condition ranges from hyperplasia to malignant lymphoid proliferation. Risk factors affecting the incidence of PTLD include allograft type, Epstein-Barr virus infection, and immunosuppression. In this article, we review the clinical, histopathologic, and imaging features of PTLD in children. Because PTLD can affect nearly any organ system, a wide variety of clinical manifestations is possible. The heterogeneous nature of the disease is also reflected on imaging studies. The goals of imaging in patients with PTLD are to detect disease, guide biopsy, and direct appropriate follow-up imaging rather than to establish a specific diagnosis. Because the clinical and imaging manifestations of PTLD are nonspecific and are not reliably predictive of histopathologic subtype, tissue biopsy is necessary for final diagnosis. PMID- 11012420 TI - Carotid artery stent placement for atherosclerotic disease: rationale, technique, and current status. AB - Carotid arterial endarterectomy is considered to be the standard for the treatment of atherosclerotic carotid arterial occlusive disease. This has been validated with results of several randomized controlled trials in which its effectiveness has been demonstrated over that of the best nonsurgical therapy. In the past several years, however, carotid angioplasty with stent placement has emerged as a potential alternative to carotid endarterectomy. This article represents a critical examination of the rationale for carotid revascularization; the history of endovascular techniques for the treatment of carotid atherosclerosis, beginning with balloon angioplasty and evolving to the use of stents; and the evidence supporting the effectiveness of the endovascular approach. A brief description of the current technical aspects of carotid artery stent placement is presented. The future status of the endovascular approach will be determined with randomized trials in which carotid artery stent placement is directly compared with endarterectomy, as well as by the potential for further innovation and improvement in endovascular devices, technique, and safety. PMID- 11012421 TI - Systemic lupus erythematosus: brain MR imaging and single-voxel hydrogen 1 MR spectroscopy. AB - PURPOSE: To evaluate the usefulness of magnetic resonance (MR) imaging and hydrogen 1 MR spectroscopy in the detection of brain involvement in patients with systemic lupus erythematosus (SLE) with or without neuropsychiatric symptoms. MATERIALS AND METHODS: Twenty-six patients who had SLE with (n = 17) or without (n = 9) neuropsychiatric symptoms were examined at MR imaging and (1)H MR spectroscopy. The voxel was placed in the basal ganglia and peritrigonal white matter. Eight healthy volunteers were included. RESULTS: Five of nine patients with major neuropsychiatric symptoms and one of eight patients with minor neuropsychiatric symptoms had abnormal MR imaging findings. (1)H MR spectroscopy showed a significantly decreased N:-acetylaspartate-creatine (Cr) ratio in the basal ganglia and an increased choline-Cr ratio in the peritrigonal white matter in patients with major symptoms compared with those with minor symptoms, those without symptoms, and healthy control subjects. Among patients with major symptoms, there was no difference in metabolite ratios between those with and those without abnormal MR imaging findings. Among patients with normal MR imaging findings, abnormal spectral changes were observed only in those with major neuropsychiatric symptoms. In patients without neuropsychiatric symptoms, results of (1)H MR spectroscopy and MR imaging were normal. CONCLUSION: In patients with SLE, (1)H MR spectroscopic findings seem to reflect the cerebral metabolic disturbance related to the severity of the neuropsychiatric symptoms and are not related to the presence of abnormal MR imaging findings. PMID- 11012422 TI - Intracranial leptomeningeal metastases: comparison of depiction at FLAIR and contrast-enhanced MR imaging. AB - PURPOSE: To compare contrast material-enhanced T1-weighted and fluid-attenuated inversion-recovery (FLAIR) magnetic resonance (MR) images in depicting leptomeningeal metastases. MATERIALS AND METHODS: Malignant lesions detected at cytologic examination of cerebrospinal fluid in 70 patients were reviewed. There were 58 studies in which both FLAIR and contrast-enhanced T1-weighted spin-echo MR images were available. A senior neuroradiologist reviewed the images from each sequence individually and separately for signs of leptomeningeal metastases and assigned a diagnostic rating of positive, indeterminate, or negative. RESULTS: Leptomeningeal metastases were depicted in 38 cases on contrast-enhanced T1 weighted spin-echo images and in 20 cases on FLAIR images. In three cases, leptomeningeal metastases were detected by using only FLAIR images. In 20 cases, leptomeningeal metastases were detected by using only contrast-enhanced T1 weighted spin-echo images. FLAIR imaging has a sensitivity of 34% for cytologically proved leptomeningeal metastases. Gadolinium-enhanced MR imaging has a sensitivity of 66%. CONCLUSION: Used alone, contrast-enhanced T1-weighted images are better than FLAIR images for detecting leptomeningeal metastases. This is particularly true for cases in which leptomeningeal metastases manifest primarily or solely as cranial nerve involvement. PMID- 11012423 TI - Lens of the eye: radiation dose in balloon dacryocystoplasty. AB - PURPOSE: To evaluate the absorbed radiation dose to the lens of the eye, which is the critical organ in the primary beam during fluoroscopically guided transluminal balloon dilation of the lacrimal drainage system (balloon dacryocystoplasty) for obstructive epiphora and to evaluate the possibility of deterministic radiation effect on the lens. MATERIALS AND METHODS: The radiation dose to the lens of the eye during balloon dacryocystoplasty (which includes pre- and postintervention dacryocystography) was measured in 10 consecutive patients by using thermoluminescent dosimeters on the lids of both eyes as close as possible to the lenses. A C-arm angiographic unit coupled with a digital imaging system was used, with similar exposure and geometric parameters in all cases. RESULTS: The mean radiation dose to the lens of the treated eye was 4.6 mGy +/- 2.2 (dose range, 1.9-9.1 mGy) and to that of the untreated eye was 38.5 mGy +/- 17.5 (dose range, 14.7-67.8 mGy). CONCLUSION: The lens of the untreated eye receives a higher dose than that of the treated eye because of its closer proximity to the x-ray tube in a lateral projection. In the lens, even the highest measured radiation dose (67.8 mGy) still was well below the deterministic threshold for lens opacity and cataract formation. PMID- 11012424 TI - Cost-effectiveness of functional imaging tests in the diagnosis of Alzheimer disease. AB - PURPOSE: To evaluate the cost-effectiveness of functional neuroimaging in the work-up of patients at specialized Alzheimer disease clinics. MATERIALS AND METHODS: A decision model was used to calculate costs and benefits (in quality adjusted life-years [QALYs]) that accrued to hypothetical cohorts of patients at presentation to an Alzheimer disease center. Sensitivity analysis was performed to examine the effects of diagnostic test characteristics, therapeutic efficacy, disease severity, and costs on cost-effectiveness. RESULTS: The incremental cost effectiveness ratio of dynamic susceptibility contrast material-enhanced magnetic resonance (MR) imaging was $479,500 per QALY (compared with the usual diagnostic work-up), while visual or quantitative single photon emission computed tomography (SPECT) was dominated (higher costs, lower effectiveness) by the usual diagnostic work-up. These results depend critically on the sensitivity and specificity of the standard diagnostic work-up, the effectiveness of drug treatment, and the disease severity. Varying these parameters resulted in estimates of incremental cost-effectiveness for dynamic susceptibility contrast-enhanced MR imaging of $24,680 to $8.6 million per QALY. SPECT either was dominated by the usual diagnostic work-up or had cost-effectiveness ratios of $180,200 to $6 million per QALY. CONCLUSION: The addition of functional neuroimaging to the usual diagnostic regimen at Alzheimer disease clinics is not cost-effective given the effectiveness of currently available therapies. PMID- 11012425 TI - A scale of methodological quality for clinical studies of radiologic examinations. AB - PURPOSE: To develop and evaluate a scale for assessment of the methodological quality for clinical investigations of radiologic studies. MATERIALS AND METHODS: A scale was developed that included methodological standards compiled from established sources for assessing the methodological quality of study designs in clinical research and characteristics related to biases commonly observed in clinical radiologic research. The scale was composed of 15 standards and was tested with the results of 96 studies on imaging of liver hemangioma. Interrater reliability was measured between two observers by using percentage agreement and kappa statistics. Interrater reliability between two observers for a composite quality index that encompassed the 15 standards was measured with the intraclass correlation coefficient. RESULTS: Agreement between the two observers was almost perfect (kappa value, 0.8-1.0) for 11 standards and substantial (kappa value, 0.74-0.78) for four standards. Agreement between the observers with regard to the composite quality index also was high (intraclass correlation coefficient r, 0.91 [95% CI: 0.87, 0.94]). CONCLUSION: The scale appears to be reliable for the assessment of methodological quality of clinical investigations of radiologic studies. PMID- 11012426 TI - Nonsurgical management of blunt splenic injury: use of CT criteria to select patients for splenic arteriography and potential endovascular therapy. AB - PURPOSE: To determine if contrast material-enhanced spiral computed tomography (CT) can be used to select patients with blunt splenic injuries to undergo arteriographic embolization. MATERIALS AND METHODS: During a 15-month period, 78 patients who were hemodynamically stable and required no immediate surgery underwent contrast-enhanced spiral CT followed by splenic arteriography. CT scans were assessed for splenic vascular contrast material extravasation or posttraumatic splenic vascular lesions. Medical records were reviewed for splenic arteriographic results and clinical outcome. RESULTS: There were 25 grade I, 12 grade II, 27 grade III, 12 grade IV, and two grade V splenic injuries. CT showed active contrast material extravasation in seven patients and splenic vascular lesions in 19 patients. At CT, splenic vascular contrast material extravasation was 100% (seven of seven patients) and a posttraumatic splenic vascular lesion was 83% (10 of 12 patients) sensitive on the basis of arteriographic or surgical outcome in predicting the need for transcatheter embolization or splenic surgery. Overall, CT had a sensitivity of 81% (17 of 21 patients), a specificity of 84% (48 of 57 patients), negative and positive predictive values of 92% (48 of 52 patients) and 65% (17 of 26 patients), respectively, and an accuracy of 83% (65 of 78 patients) in predicting the need for splenic injury treatment. CONCLUSION: Contrast-enhanced spiral CT plays a valuable role in selecting hemodynamically stable patients with splenic vascular injury who may be treated with transcatheter therapy and potentially improves the success rate of nonsurgical management. PMID- 11012427 TI - US vascular mapping before hemodialysis access placement. AB - PURPOSE: To prospectively assess the effect of preoperative ultrasonographic (US) mapping on surgical selection, placement of arteriovenous fistulas (AVFs) and grafts, and negative surgical exploration rates. MATERIALS AND METHODS: US assessment of the upper extremity arterial and venous anatomy was performed in 70 patients with chronic renal failure before surgical evaluation. The surgeon documented the planned access procedure, which was based on physical examination results, and then reviewed the US preoperative mapping report. The surgical procedure and outcome were recorded. RESULTS: Fifty-two of the 70 patients who underwent mapping had vascular access placement. Preoperative US mapping resulted in a change in the planned surgical procedure in 16 (31%) of the 52 patients. An AVF rather than the planned graft was placed in eight (15%) patients. The AVF placement rate increased from 32% (126 of 395 patients) to 58% (30 of 52 patients). Unsuccessful surgical explorations decreased from 11% (28 of 256) to 0%. CONCLUSION: Preoperative US mapping before hemodialysis access placement can result in a change in surgical management, with an increased number of AVFs placed and an improved likelihood of selecting the most functional vessels preoperatively. Further study is needed to determine longer term outcomes. PMID- 11012428 TI - Tunneled infusion catheters: increased incidence of symptomatic venous thrombosis after subclavian versus internal jugular venous access. AB - PURPOSE: To compare the incidence of symptomatic venous thrombosis after tunneled infusion catheter placement via the internal jugular vein (IJV) versus the subclavian vein (SCV). MATERIALS AND METHODS: A retrospective analysis was performed of 774 catheters placed. Only patients with complete follow-up were included, which yielded a population of 279 catheters in 238 patients (166 in the SCV, 113 in the IJV; total of 26,242 catheter days). All catheters were placed by interventional radiologists with ultrasonographic (in IJV) or venographic (in SCV) guidance. RESULTS: Initial complications were limited to one pneumothorax in the SCV group and one episode of oversedation in the IJV group. There was no difference in infection rates between the two sites (SVC vs IJV: 0.25 vs 0.32 per 100 catheter days; P >.99). The mean dwell time was slightly longer for SCV catheters (103 days) than for IJV catheters (79 days) (P =.04). Venous thrombosis developed in 13% of patients (0.12 per 100 catheter days) with an SVC catheter placed as compared with in 3% (0.04 per 100 catheter days) with an IJV catheter (P =.018). This difference persisted after adjustment for catheter size and side of placement (P =.025). The mean time to thrombosis was 36 days for SCV catheters and 142 days for IJV catheters. CONCLUSION: The IJV is the preferred site for tunneled infusion catheter placement because of the lower incidence of symptomatic venous thrombosis. PMID- 11012429 TI - Peripheral arterial obstruction: prospective study of treatment with a transluminally placed self-expanding stent-graft. International Trial Study Group. AB - PURPOSE: To evaluate the safety and efficacy of an endoluminal prosthesis for treatment of peripheral arterial occlusive disease (PAOD). MATERIALS AND METHODS: A self-expanding endoprosthesis with an expanded polytetrafluoroethylene tube inside a nitinol support structure was implanted in 127 patients with symptomatic PAOD in the iliac (61 limbs) and femoral arteries (80 limbs). Clinical category status, ankle-brachial index, and color duplex flow imaging results were recorded before treatment, at discharge, and at 1, 3, 6, and 12 months after treatment. Aspirin was administered throughout the study, and heparin was administered during and for 2 days after the procedure. RESULTS: Endoprosthesis deployment was technically successful in all patients. Complications occurred in 24 of 141 procedures and included three major complications. Early thrombosis (within 30 days) occurred in one iliac and three femoral arteries. Late restenosis or reocclusion was observed in five iliac and 14 femoral arteries within the 1st year. Primary patency rates in iliac arteries were 98% +/- 3% (standard error) and 91% +/- 4%, respectively, at 6 and 12 months after treatment. Primary patency rates in femoral arteries were 90% +/- 3% and 79% +/- 5%, respectively, at 6 and 12 months. Secondary patency rates were 95% and 93% for iliac and femoral arteries, respectively, at 12 months after treatment. CONCLUSION: The device used in this study can be implanted without additional risks to the patient and provided encouraging patency rates up to 1 year. PMID- 11012430 TI - Peripheral arterial disease: meta-analysis of the diagnostic performance of MR angiography. AB - PURPOSE: To summarize the overall diagnostic performance of magnetic resonance (MR) angiography in the evaluation of peripheral arteriosclerotic occlusive disease and to identify the most important sources of variation in diagnostic accuracy between studies. MATERIALS AND METHODS: A search strategy in MEDLINE and citation tracking were used to identify relevant English-language articles published since 1991. Each article was critically appraised for examination, patient, and study design characteristics. The accuracy data from different studies were analyzed by constructing summary receiver operating characteristic curves; multiple linear regression was used to examine the variation between study results. RESULTS: Twenty-three studies were included. There was much heterogeneity in the study results, which could not be explained as differences in the threshold for a positive result. About half of the variation was due to the type of MR angiographic examination and the extent of image evaluation. The relative diagnostic odds ratio (DOR) for three-dimensional (3D) gadolinium enhanced MR angiography compared with two-dimensional (2D) time-of-flight MR angiography was 7.46 (95% CI: 2.48, 22.20). The relative DOR for review of transverse source images or multiplanar reformations in addition to maximum intensity projections (MIPs) compared with the use of only MIPs for image evaluation was 4.53 (95% CI: 1.46, 13.87). CONCLUSION: The diagnostic accuracy of 3D gadolinium-enhanced MR angiography is superior to that of 2D time-of-flight MR angiography. Also, the review of transverse source images or use of additional postprocessing techniques, such as multiplanar reformation, results in significantly better diagnostic performance. PMID- 11012431 TI - Suppression of intravascular signal on fat-saturated contrast-enhanced thoracic MR arteriograms. AB - PURPOSE: To assess the prevalence of artifactual signal intensity loss within the aortic arch and proximal branch vessels on fat-saturated contrast material enhanced magnetic resonance (MR) arteriograms of the thoracic aorta and to hypothesize about the cause of the loss of signal intensity. MATERIALS AND METHODS: Between January and June 1998, 105 consecutive MR arteriograms of the thoracic aorta were acquired in 103 patients at 1.5 T. Imaging included an arterial phase three-dimensional (3D) fat-saturated contrast-enhanced gradient echo (GRE) sequence followed by a delayed two-dimensional (2D) transverse fat saturated GRE sequence. All MR images were reviewed by two radiologists who were blinded to patient history and results of imaging studies and who evaluated the images for the presence of intraluminal loss of signal intensity in the aortic arch and the proximal branch vessels. RESULTS: Intravascular loss of signal intensity was present in at least one vessel on 23 of the 105 arterial phase 3D studies. Seventy-one of 91 left subclavian arterial segments had loss of signal intensity on the delayed 2D studies. CONCLUSION: Intravascular signal intensity loss can be present on contrast-enhanced fat-saturated images of the aortic arch and proximal branch vessels, particularly the left subclavian artery. This phenomenon, which is to the authors' knowledge previously unreported and which is hypothesized to result from undesired water saturation, should not be misinterpreted as stenotic or occlusive vascular disease. PMID- 11012432 TI - Percutaneous radio-frequency thermal ablation of nonresectable hepatocellular carcinoma after occlusion of tumor blood supply. AB - PURPOSE: To evaluate the usefulness of percutaneous radio-frequency (RF) thermal ablation of nonresectable hepatocellular carcinoma (HCC) after occlusion of the tumor arterial supply. MATERIALS AND METHODS: Sixty-two patients with cirrhosis and biopsy-proved HCC underwent RF ablation after interruption of the tumor arterial supply by means of occlusion of either the hepatic artery with a balloon catheter (40 patients) or the feeding arteries with gelatin sponge particles (22 patients). RESULTS: After a single RF procedure in 56 patients and after two procedures in six patients, spiral computed tomography (CT) demonstrated a nonenhancing area corresponding in shape to the previously identified HCC, which was suggestive of complete necrosis. No major complications occurred. Two patients subsequently underwent surgical resection; the remaining 60 patients were followed up with spiral CT. During a mean follow-up of 12.1 months, 11 HCC nodules showed areas of local progression; 49 were identified as nonenhancing areas with a 40%-75% reduction in maximum diameter. The 1-year estimate of failure risk was 19% for local recurrence and 45% for overall intrahepatic recurrence. The estimated 1-year survival was 87%. Histopathologic analysis of one autopsy and two surgical specimens revealed more than 90% necrosis in one specimen and 100% necrosis in two. CONCLUSION: HCC nodules 3.5-8.5 cm in diameter can be ablated in one or two RF sessions after occlusion of the tumor arterial supply. PMID- 11012433 TI - Serum ethanol levels in children and adults after ethanol embolization or sclerotherapy for vascular anomalies. AB - PURPOSE: To evaluate the serum ethanol level in children and adults after ethanol embolization or sclerotherapy for vascular anomalies. MATERIALS AND METHODS: Serum ethanol level was evaluated in 71 patients with vascular anomalies who underwent ethanol embolization, sclerotherapy, or both. Blood used to determine serum ethanol level was drawn at the end of each procedure. RESULTS: The relationship between serum ethanol level and amount of ethanol administered was statistically significant. Results of stepwise linear regression analysis showed that the amount of ethanol administered (P <.001) and type of malformation (P =.02) were multivariate predictors of serum ethanol level. Results of logistic regression analysis revealed that the only uni- or multivariate predictor of legal intoxication was the amount of ethanol administered (P =.018). Five patients fulfilled the criteria for legal intoxication, and the mean amount of ethanol administered to these patients was 0.87 mL per kilogram of body weight +/ 0.18 (SD) (range, 0.55-0.99 mL/kg). CONCLUSION: The volume of ethanol administered is the most reliable predictor of serum ethanol level and legal intoxication. Patients who receive up to 1.0 mL/kg ethanol during embolization or sclerotherapeutic procedures may have elevated serum ethanol levels that could put them at risk of respiratory depression, cardiac arrhythmias, seizures, rhabdomyolysis, and hypoglycemia. PMID- 11012434 TI - Case 31 PMID- 11012435 TI - Case 27: intrapancreatic accessory spleen. PMID- 11012436 TI - Heterotopic pancreas in the stomach: CT findings. AB - PURPOSE: To describe the computed tomographic (CT) findings of heterotopic pancreas in the stomach. MATERIALS AND METHODS: CT findings in 12 patients with heterotopic pancreas in the stomach were reviewed. Surgical resection (n = 11) or endoscopic excision (n = 1) was performed in cases of symptomatic heterotopic pancreas (n = 4), suspected submucosal tumors (n = 7), and gastric carcinoma (n = 1). Seven patients underwent helical CT with water as an oral contrast agent; five underwent nonhelical CT with water-soluble contrast material. RESULTS: Nine heterotopic pancreata were in the antrum and one each was in the body, fundus, and perigastric fat. Seven lesions were on the greater curvature aspect; five, on the lesser curvature aspect. Common CT findings were well-defined oval or round masses with smooth or serrated margins in the gastric antral wall. Four of the seven lesions in which helical CT was performed enhanced similarly to normal pancreas. Preoperatively, CT depicted 11 of the 12 lesions, but CT findings were interpreted correctly as heterotopic pancreas in only two; the remaining 10 were misinterpreted as other lesions. Atypical findings were cystic dilatation of heterotopic pancreatic duct in two, unusual location in the fundus or perigastric fat in two, and malignant transformation in one. CONCLUSION: CT findings of heterotopic pancreas in the stomach appear to be nonspecific for diagnosis, except for location. PMID- 11012437 TI - Fibrolamellar hepatocellular carcinoma: pre- and posttherapy evaluation with CT and MR imaging. AB - PURPOSE: To determine the features of advanced hepatic and extrahepatic fibrolamellar hepatocellular carcinomas (HCCs) and their effects on immediate surgical management and tumor recurrence. MATERIALS AND METHODS: Thirty-one patients with fibrolamellar HCC underwent pretherapy computed tomography (CT); 11 underwent pretherapy magnetic resonance (MR) imaging. All 40 patients underwent posttherapy CT; four, follow-up MR imaging. Imaging, surgical, and histopathologic findings were correlated. RESULTS: Twenty-five (81%) patients had solitary tumors (mean maximum diameter, 13 cm). Thirteen (42%) patients had intrahepatic biliary obstruction; 27 (87%) patients had involvement of the portal or hepatic veins. Thirteen (42%) had extrahepatic tumor spread, nine (29%) had distant metastases on pretherapy images, and 20 (65%) had lymphadenopathy. Thirty two (80%) of 40 patients underwent exploration surgery; curative resection was attempted in 25 (62%), including four patients who underwent liver transplantation. Only 17 patients were considered to have had hepatic and extrahepatic tumors completely excised. Tumor recurred in all eight of the 17 patients who had extrahepatic disease at pretherapy CT and in four of the seven patients who seemed to have tumor limited to the liver. A combination of repeat tumor resection and adjuvant chemotherapy resulted in prolonged tumor-free survival in some cases. CONCLUSION: Fibrolamellar HCC frequently demonstrates aggressive local invasion and nodal and distant metastases. Pretherapy and follow up imaging are important for staging, surveillance, and optimal management. Aggressive surgical resection may be helpful to control fibrolamellar HCC and to prolong survival in appropriately selected cases. PMID- 11012438 TI - Hepatic lesion detection and characterization: value of nonenhanced MR imaging, superparamagnetic iron oxide-enhanced MR imaging, and spiral CT-ROC analysis. AB - PURPOSE: To determine the accuracy for detection and characterization of focal hepatic lesions of nonenhanced, superparamagnetic iron oxide (SPIO)-enhanced, or a combination of nonenhanced and SPIO-enhanced MR imaging and contrast-enhanced spiral computed tomography (CT). MATERIALS AND METHODS: Spiral CT and T2-weighted SPIO-enhanced (ferucarbotran-enhanced) MR imaging were performed in 35 patients within 2 weeks before surgery for malignant hepatic lesions. Only malignant lesions with histopathologic proof were considered. A total of 875 images with and 800 images without focal lesions were presented to five readers, who were asked to assess the presence and characterization of lesions by using a five point confidence scale. Receiver operating characteristic analysis was performed. RESULTS: Nonenhanced and SPIO-enhanced images together and SPIO-enhanced images alone yielded the best performance for lesion detection. No differences were found among all imaging techniques with regard to lesion characterization (benign vs malignant). The combined approach resulted in larger area under the ROC curve (A(z) = 0.9062) and accuracy (85.3%) (P < 0.02), as compared with SPIO-enhanced MR imaging (A(z) = 0.8667; accuracy, 73.1%). CONCLUSION: SPIO-enhanced T2 weighted MR imaging was more accurate than nonenhanced T1-weighted and T2 weighted MR imaging and contrast-enhanced spiral CT for the detection of focal hepatic lesions. The combined analysis of nonenhanced and SPIO-enhanced images was more accurate in the characterization of focal hepatic lesions than was review of SPIO-enhanced images alone. PMID- 11012439 TI - Right lower quadrant pain and suspected appendicitis: nonfocused appendiceal CT- review of 100 cases. AB - PURPOSE: To determine the value of standard, nonfocused computed tomography (CT) in examining patients with right lower quadrant (RLQ) pain and suspected appendicitis. MATERIALS AND METHODS: The CT scans and medical records of 100 consecutive patients who presented to the emergency department with RLQ pain and were clinically suspected of having appendicitis were retrospectively reviewed. Helical CT of both the abdomen and pelvis was performed at 7-mm increments after oral and intravenous contrast material administration. CT scans were evaluated for the presence of appendiceal or other disease. Results were correlated with surgical and pathologic findings in 34 patients or with 3-month clinical follow up in 66 patients. RESULTS: CT depicted abnormalities in 66 patients (66%). In 59 (59%) patients, the abnormality was located in the pelvis; 23 (39%) of these patients had appendicitis. Seven (7%) patients had abnormalities outside of the pelvis, a region not typically scanned during focused appendiceal imaging. Four of these seven patients required surgery. Thus, if only pelvic focused RLQ CT had been performed, overall sensitivity would have decreased from 99% to 88% (P <.05) and sensitivity for cases necessitating surgery would have decreased from 96% to 82% (P <.05). CONCLUSION: Both abdominal and pelvic CT examinations are necessary to increase sensitivity and identify the many possible causes of RLQ pain in patients with clinically suspected appendicitis. PMID- 11012440 TI - Influence of image acquisition parameters on CT artifacts and polyp depiction in spiral CT colonography: in vitro evaluation. AB - PURPOSE: To quantify the effects of spiral computed tomographic (CT) acquisition parameters on the magnitude of three-dimensional (3D) rippling artifacts and polyp depiction. MATERIALS AND METHODS: An in vitro colon phantom was constructed with air-filled acrylic cylinders that contained synthetic polyps of 3-13 mm. The phantom was submerged in fluid and positioned at four angles of inclination relative to the z axis. Image data were acquired at collimation and pitch combinations of 3 mm and 1.67 and 5 mm and 1.6, respectively. Rippling artifacts were quantified by measuring the longitudinal variation of in-plane phantom edge width, and the influence of these artifacts on the depiction of pedunculated and sessile polyps was assessed qualitatively. RESULTS: The in-plane magnitude of the rippling artifact was a function of the angle of inclination relative to the longitudinal axis and the table increment. The through-plane periodicity of the artifact was equal to one-half the table increment. CONCLUSION: The table increment and angle of inclination of the surface of the object relative to the z axis determine the periodicity and magnitude of the rippling artifact at 3D spiral CT colonography. Although the depiction of small pedunculated polyps was not compromised, some sessile polyps were degraded by the artifact. PMID- 11012441 TI - Stent appearance at contrast-enhanced MR angiography: in vitro examination with 14 stents. AB - PURPOSE: To evaluate signal intensity changes influencing assessment of stent patency at contrast material-enhanced magnetic resonance (MR) angiography. MATERIALS AND METHODS: By using an in vitro model, 14 stents-nine nitinol, one tantalum, two stainless steel, and two cobalt alloy-were investigated regarding their appearance at MR imaging. A vascular phantom consisting of tubes filled with 2.00 mmol/L gadopentetate dimeglumine in saline solution was studied in different orientations within the magnetic field. Imaging was performed with a fast three-dimensional gradient-echo sequence (4. 70/1.89 [repetition time msec/echo time msec]). Relative signal intensity reduction within the stents and the degree of artificial narrowing of the stent lumen were calculated. RESULTS: The stent lumen was visible within 13 stents. A total signal void inside the stent lumen appeared in only one cobalt alloy stent. Artificial narrowing of the diameter was less than 33% in 10 of 14 stents. The tantalum stent and four nitinol stents seemed best suited for contrast-enhanced MR angiography. A bandlike artifact occurred at the ends of the stents when positioned along the readout direction. CONCLUSION: To differentiate between artifacts and stenoses, knowledge of the degree of signal intensity reduction and artificial lumen narrowing within vascular stents is essential. Stent geometry, relative orientation to the magnetic field, and alloy composition influence signal intensity alteration within the stent lumen. PMID- 11012442 TI - CT of the middiaphyseal femur: cortical bone mineral density and relation to porosity. AB - PURPOSE: To determine whether computed tomography (CT) can be used to quantify age- and site-related changes in cortical bone mineral density (cBMD) at the middiaphyseal femur and whether cBMD differences are related to intracortical porosity. MATERIALS AND METHODS: Cortical bone specimens from 163 femurs were studied with CT and microradiography. Femurs were from 77 males and 86 females in a white anthropologic collection covering a broad age spectrum. In each sample, the cBMD was measured in the entire cortical width and in periosteal, midcortical, and endosteal subregions of interest. Age- and site-related changes in cBMD were tested for significance by using a two-way analysis of variance for both sexes. By using linear regression, cBMD was compared with porosity in the entire cortical width and in each subregion. RESULTS: There were significant age related differences in cBMD (P <.001 in females, P =.008 in males). In addition, cBMD values were significantly different between the three cortical subregions (P <.001 for both sexes), decreasing from the periosteum to the midcortex to the endosteum. The cBMD values were closely related to porosity, and porosity contributed to 71.6% of the variance in cBMD in the overall population. CONCLUSION: CT is effective in the measurement of age- and site-related changes in cBMD. Decreases in cBMD are closely correlated with increased cortical porosity. PMID- 11012443 TI - Femoral head osteonecrosis: detection and grading by using a rapid MR imaging protocol. AB - PURPOSE: To design and evaluate a limited magnetic resonance (MR) imaging examination that can be performed rapidly and potentially inexpensively in patients with clinical suspicion of osteonecrosis. MATERIALS AND METHODS: Both a limited and a full hip MR examination were performed prospectively in 179 hips in 92 patients with clinical suspicion of femoral head osteonecrosis. The presence of osteonecrosis was determined by two radiologists. The percentage of involvement of the femoral head weight-bearing surface was evaluated subsequently for osteonecrosis-positive hips on both sets of images. RESULTS: Both examinations were performed successfully in all cases. Agreement between the limited and full examinations for presence of osteonecrosis was 98.9% (177 of 179 cases; kappa, 0.97). Forty-six (92%) of 50 patients with femoral head osteonecrosis at both examinations were placed in the appropriate quartile of percentage of femoral head weight-bearing surface involvement by both readers (weighted kappa, 0.94). Incidental findings were made at the full examination that could not be made or were difficult to make at the limited examination. CONCLUSION: There was excellent agreement between the full and screening MR examinations for both detection of and determining the extent of osteonecrosis. The time and potential cost reduction achieved with a limited examination may allow introduction of MR imaging earlier in the diagnosis of femoral head osteonecrosis, as well as its more widespread use in patient care. PMID- 11012444 TI - Characterization of the "red zone" of knee meniscus: MR imaging and histologic correlation. AB - PURPOSE: To determine the extent and vascularity of knee menisci with conventional and gadolinium-enhanced magnetic resonance (MR) imaging in cadaveric specimens, with histologic findings as the reference standard, and to investigate signal intensity changes in menisci and perimeniscal soft tissues in symptomatic patients. MATERIALS AND METHODS: Radial dimensions and enhancement patterns of menisci were recorded and compared in (a) 12 cadaveric menisci examined with conventional and gadolinium-enhanced intermediate-weighted and fat-suppressed T1 weighted spin-echo MR imaging, high-spatial-resolution T1-weighted and fast low angle shot MR imaging, and gross anatomic and histologic specimens and (b) 18 patients examined with conventional and gadolinium-enhanced fat-suppressed T1 weighted spin-echo MR imaging. RESULTS: No differences in radial measurements of the meniscus were found for different MR techniques (P =.551). Despite the presence of vessels in the peripheral 10%-15% of the menisci, no enhancement of menisci was detected in specimens or patients. Perimeniscal soft-tissue enhancement adjacent to the posterior horn was greater than that adjacent to the anterior horn (P <.05), and enhancement of the lateral meniscal body was greater than that of the medial meniscal body (P <.05). CONCLUSION: The wedge-shaped low signal-intensity structure seen on MR images represents the entire meniscus. Intravenous injection of contrast material does not appear to be useful for differentiation of the vascularized from the nonvascularized zone of the meniscus. PMID- 11012445 TI - Pulley system in the fingers: normal anatomy and simulated lesions in cadavers at MR imaging, CT, and US with and without contrast material distention of the tendon sheath. AB - PURPOSE: To describe the normal anatomy of the finger flexor tendon pulley system, with anatomic correlation, and to define criteria to diagnose pulley abnormalities with different imaging modalities. MATERIALS AND METHODS: Three groups of cadaveric fingers underwent computed tomography (CT), magnetic resonance (MR) imaging, and ultrasonography (US). The normal anatomy of the pulley system was studied at extension and flexion without and with MR tenography. Pulley lengths were measured, and anatomic correlation was performed. Pulley lesions were created and studied at flexion, extension, and forced flexion. Two radiologists reviewed the studies in blinded fashion. RESULTS: MR imaging demonstrated A2 (proximal phalanx) and A4 (middle phalanx) pulleys in 12 (100%) of 12 cases, without and with tenography. MR tenography showed the A3 (proximal interphalangeal) and A5 (distal interphalangeal) pulleys in 10 (83%) and nine (75%) cases, respectively. US showed the A2 pulley in all cases and the A4 pulley in eight (67%). CT did not allow direct pulley visualization. No significant differences in pulley lengths were measured at MR, US, or pathologic examination (P: =.512). Direct lesion diagnosis was possible with MR imaging and US in 79%-100% of cases, depending on lesion type. Indirect diagnosis was successful with all methods with forced flexion. CONCLUSION: MR imaging and US provide means of direct finger pulley system evaluation. PMID- 11012446 TI - The musculoskeletal crescent sign. PMID- 11012447 TI - Rectal bleeding after radiation therapy for prostate cancer: endoscopic evaluation. AB - PURPOSE: To analyze the frequency and clinical importance of proctitis and hematochezia after radiation therapy for prostate cancer. MATERIALS AND METHODS: Of 63 patients with prostate cancer treated with curative intent by a single radiation oncologist between July 1, 1993, and December 31, 1997, 30 were asymptomatic, but 33 had heme-positive digital rectal examination (DRE) results or hematochezia at routine follow-up. Twenty-six of these patients underwent endoscopy of the sigmoid colon or colon for evaluation of these symptoms. Median doses of 60.0 Gy at postoperative radiation therapy and 68.4 Gy at definitive radiation therapy were delivered to four fields daily by using blocking customized on the basis of computed tomographically documented evidence of disease. The Fisher exact test and the Kaplan-Meier method were used to analyze the results. RESULTS: The frequency of rectal bleeding approached 80% at 3 years after radiation therapy in definitively treated patients. Only 14 patients had proctitis: eight as the only sign, and six in association with other disease. Six patients had other disease without proctitis, and four patients had normal examination findings. The frequency of rectal bleeding in the presence of proctitis was similar to that in the presence of other disease (Fisher exact test, P =.68). CONCLUSION: Hematochezia or positive DRE findings are frequent sequelae of definitive radiation therapy for prostate cancer; however, causes other than proctitis are often documented at endoscopy. Symptomatic individuals warrant rigorous evaluation to rule out serious coexistent disease. PMID- 11012448 TI - Scrotal disorders: evaluation of testicular enhancement patterns at dynamic contrast-enhanced subtraction MR imaging. AB - PURPOSE: To evaluate testicular enhancement patterns in various scrotal disorders at dynamic contrast medium-enhanced subtraction magnetic resonance (MR) imaging. MATERIALS AND METHODS: Forty-two patients with scrotal symptoms (22 testicular diseases, 20 extratesticular scrotal disorders) underwent three-dimensional (3D) fast field-echo or fast spin-echo dynamic subtraction MR imaging after injection of paramagnetic contrast medium. The relative percentages of peak height and mean slope of the testes on the affected side were compared with those on the unaffected side by using time-signal intensity curves. RESULTS: Extratesticular scrotal disorders (time-signal intensity curve mean peak height, 93.1%; mean slope, 89.8%) showed gradual and progressive increase in homogeneous testicular contrast enhancement in all normal testes. Relative percentages of peak height and mean slope of testicular torsion (mean peak height, 17.3%; mean slope, 10.6%), infarction (mean peak height, 30.4%; mean slope, 19.8%), traumatic hemorrhagic necrosis (mean peak height, -3.5%; mean slope, -12.0%), and epidermoid cyst (mean peak height, -6.6%; mean slope, -14.2%) were significantly lower than those of extratesticular scrotal disorders. Acute mumps orchitis (mean peak height, 135.1%; mean slope, 307.5%) and malignant testicular tumor (mean peak height, 178.7%; mean slope, 467.6%) showed higher relative percentages of peak height and mean slope. CONCLUSION: Dynamic contrast-enhanced subtraction MR imaging can provide information about testicular perfusion on the basis of contrast enhancement and can be used to differentiate testicular diseases from scrotal disorders. PMID- 11012449 TI - Symptomatic fibroleiomyomata: MR imaging of the uterus before and after uterine arterial embolization. AB - PURPOSE: To determine the magnetic resonance (MR) imaging features of uterine fibroleiomyomata after uterine arterial embolization (UAE) and identify pretreatment MR imaging features that may be predictive of successful UAE. MATERIALS AND METHODS: T1- and T2-weighted and dynamic gadolinium-enhanced T1 weighted images were obtained before and 3 months after UAE in 31 patients. Up to five fibroleiomyomata (total of 125) were evaluated for volume, location, signal intensity characteristics, and vascularity. Region-of-interest curves were used to assess the vascular enhancement pattern of each fibroleiomyoma and adjacent myometrium. Each patient completed a questionnaire on symptoms 3 months after UAE. RESULTS: UAE resulted in significant reductions in mean uterine volume (from 588.6 to 393.1 cm(3)) and mean fibroleiomyoma volume (from 69.4 to 41.4 cm(3)) (P <.005). After UAE, lesions showed signal intensity changes consistent with hemorrhagic infarction. The vascularity of fibroleiomyomata was decreased (P <.001), with no significant change in myometrial vascularity. Submucosal location was a strong positive predictor of fibroleiomyoma volume reduction (P < 001). When a reduction in vascularity was the measure of success, hypervascularity was a strong indicator of success (P <. 005). CONCLUSION: MR imaging is useful for quantitative assessment of signal intensity and morphologic changes before and after UAE. Pretreatment MR imaging findings may help predict the success of the procedure. PMID- 11012450 TI - Fetal lip and primary palate: three-dimensional versus two-dimensional US. AB - PURPOSE: To determine if three-dimensional (3D) ultrasonography (US) improves the ability to define the location and extent of facial clefting prenatally compared with two-dimensional (2D) US. MATERIALS AND METHODS: Thirty-one fetuses suspected of having a facial cleft were examined prospectively with 2D and 3D US. Follow-up was performed in all fetuses. RESULTS: Twenty-eight fetuses had a cleft lip at birth. The location of the cleft lip was correctly identified in all fetuses with 3D US and in 26 of 28 with 2D US. Twenty-two fetuses had a cleft primary palate. Nineteen and nine of 22 cleft palates were identified by using 3D and 2D US, respectively. Three fetuses suspected of having a facial cleft at 2D US had a normal palate at 3D US and at birth. CONCLUSION: Three-dimensional US is useful to identify the location and extent of facial clefting. The advantages of 3D US are the following: (a) The face may be viewed in a standard orientation, (b) the defect may be viewed systematically by using an interactive display, and (c) the rendered image provides landmarks for the planar images. Patient decisions may be affected, since they can view the abnormality on a recognizable 3D rendered image. PMID- 11012451 TI - Nonpalpable breast lesions: evaluation with power Doppler US and a microbubble contrast agent-initial experience. AB - PURPOSE: To evaluate power Doppler ultrasonography (US) performed with a microbubble US contrast agent in the differentiation of nonpalpable breast lesions. MATERIALS AND METHODS: Fifty nonpalpable breast lesions in 50 patients were prospectively evaluated with power Doppler US before and after injection of the contrast agent SH U 508A. Lesion vascularity and the morphology of vessels on US scans were analyzed and were correlated with histologic results. RESULTS: Surgical excision revealed 22 cancers and 28 benign lesions. At nonenhanced power Doppler US, eight (36%) of 22 cancers and four (14%) of 28 benign lesions were vascular. At contrast agent-enhanced power Doppler US, 21 (95%) cancers and six (21%) benign lesions were vascular (P <.001). Irregular vessels were seen in three cancers and one benign lesion at nonenhanced power Doppler US and in 11 cancers and one benign lesion at contrast-enhanced power Doppler US. By using the presence of vascularity in the mass as the diagnostic criterion for malignancy, the sensitivity, specificity, and positive and negative predictive values of power Doppler US changed from 36%, 86%, 67%, and 63%, respectively, to 95%, 79%, 78%, and 96% after contrast agent injection. CONCLUSION: Contrast-enhanced power Doppler US was superior to nonenhanced power Doppler US in the demonstration and characterization of tumor vascularity in nonpalpable breast lesions. Contrast enhanced power Doppler US may be useful for the differentiation between nonpalpable breast cancers and benign tumors. PMID- 11012452 TI - New bilateral microcalcifications at mammography in a postlactational woman: case report. AB - A 33-year-old woman with a strong family history of breast cancer who was referred for mammography 5 weeks after completing lactation was found to have new diffuse bilateral microcalcifications in the breast ducts. Contrast material enhanced magnetic resonance imaging of the breast showed bilateral patchy areas of abnormal enhancement. Large-core needle biopsy showed diffuse calcifications within expanded benign ducts in a background of lactational change, without evidence of malignancy. To the authors' knowledge, these calcifications have not been previously reported and are possibly related to milk stasis or apoptosis associated with lactation. PMID- 11012453 TI - Small pulmonary nodules: volumetrically determined growth rates based on CT evaluation. AB - PURPOSE: To determine the accuracy of high-resolution computed tomographic (CT) volumetric measurements of small pulmonary nodules to assess growth and malignancy status. MATERIALS AND METHODS: The accuracy of three-dimensional (3D) image extraction and isotropic resampling techniques was assessed by performing three experiments. The first experiment measured volumes in spherical synthetic nodules of two diameters (3.20 and 3.96 mm), the second measured deformable silicone synthetic nodules prior to and after their shape had been altered markedly, and the third measured nodules of various shapes and sizes. Three dimensional techniques were used to assess growth in 13 patients for whom the final diagnosis was known and whose initial nodule diameters were less than 10 mm. By using the exponential growth model and the calculated nodule volume at two points in time, the doubling time for each subject was calculated. RESULTS: The three synthetic nodule studies revealed that the volume could be measured accurately to within +/-3%. All five malignant nodules grew, and all had doubling times less than 177 days. Some malignant nodules had asymmetric patterns of growth identified by using the 3D techniques but not the two-dimensional methods. All eight benign nodules had doubling times of 396 days or greater or showed a decrease in volume. CONCLUSION: CT volumetric measurements are highly accurate for determining volume and are useful in assessing growth of small nodules and calculating their doubling times. PMID- 11012454 TI - Solitary pulmonary nodules in patients with extrapulmonary neoplasms. AB - PURPOSE: To determine the frequency of single lung metastasis, primary lung cancer, and benign lesions in patients with a solitary lung nodule and a primary extrapulmonary neoplasm. MATERIALS AND METHODS: The authors evaluated the electronic charts of 149 patients with an extrapulmonary malignant neoplasm and a solitary pulmonary nodule. The histologic characteristics of the nodule were correlated with those of the extrapulmonary neoplasm and with patient age and smoking history. RESULTS: Patients with carcinomas of the head and neck, bladder, breast, cervix, bile ducts, esophagus, ovary, prostate, or stomach were more likely to have primary bronchogenic carcinoma than lung metastasis (ratio, 25:3 for patients with head and neck cancers; 26:8 for patients with other types of cancer combined). Patients with carcinomas of the salivary glands, adrenal gland, colon, parotid gland, kidney, thyroid gland, thymus, or uterus had fairly even odds (ratio, 13:16). Patients with melanoma, sarcoma, or testicular carcinoma were more likely to have a solitary metastasis than a bronchogenic carcinoma (ratio, 23:9). Thirty patients had a benign nodule. There was substantial overlap in age distribution among the patients with benign disease, lung cancer, and metastasis, although no patient younger than 44 years had a lung cancer. Smokers had a 3.5-fold higher chance of developing lung cancer compared with nonsmokers. CONCLUSION: The likelihood of a primary lung cancer versus a metastasis depends on the histologic characteristics of the extrapulmonary neoplasm and the patient's smoking history. PMID- 11012455 TI - Infectious pseudoaneurysms suspected at echocardiography: electron-beam CT findings. AB - PURPOSE: To review the electron-beam computed tomographic (CT) findings in patients with clinical endocarditis and suspected of having perivalvular pseudoaneurysms at echocardiography and to compare these findings with echocardiographic data. MATERIALS AND METHODS: Data on 17 patients who underwent electron-beam CT for suspicion of perivalvular infectious pseudoaneurysm at echocardiography were retrospectively reviewed. Thirteen patients had a history of valvular surgery. Electron-beam CT findings-lesion size, number, extent, and relationships with surrounding structures, and associated lesions-were compared with echocardiographic and surgical and/or autopsy data. RESULTS: In all patients, electron-beam CT depicted one or more abnormal cavities that filled with contrast material after bolus injection. The mean size (3.5 cm) and number (n = 21) of pseudoaneurysms recorded with electron-beam CT were greater than those recorded with echocardiography (2.9 cm and n = 13, respectively). Associated electron-beam CT findings included valvular vegetations in three patients; mediastinitis in two; and coronary arterial involvement in six. In eight (47%) patients, electron-beam CT depicted a pseudoaneurysm or an additional pseudoaneurysm that was only suspected-not depicted-at echocardiography. Transthoracic and transesophageal echocardiography resulted in underestimation of lesion number, size, and extent and associated lesions, particularly in patients with valvular prostheses or voluminous lesions. CONCLUSION: Thoracic infectious pseudoaneurysms are well depicted with electron-beam CT, which may be a useful addition to echocardiography for detection of this disease and thus help in preoperative planning. PMID- 11012456 TI - MR coronary angiography with breath-hold targeted volumes: preliminary clinical results. AB - PURPOSE: To assess the clinical value of a magnetic resonance (MR) coronary angiography strategy involving a small targeted volume to image one coronary segment in a single breath hold for the detection of greater than 50% stenosis. MATERIALS AND METHODS: Thirty-eight patients referred for elective coronary angiography were included. The coronary arteries were localized during single breath-hold, three-dimensional imaging of the entire heart. MR coronary angiography was then performed along the major coronary branches with a double oblique, three-dimensional, gradient-echo sequence. Conventional coronary angiography was the reference-standard method. RESULTS: Adequate visualization was achieved with MR coronary angiography in 85%-91% of the proximal coronary arterial branches and in 38%-76% of the middle and distal branches. Overall, 187 (69%) of 272 segments were suitable for comparison between conventional and MR coronary angiography. The diagnostic accuracy of MR coronary angiography for the detection of hemodynamically significant stenoses was 92%; sensitivity, 68%; and specificity, 97%. The sensitivity in individual segments was 50%-77%, whereas the specificity was 94%-100%. CONCLUSION: Adequate visualization of the major coronary arterial branches was possible in the majority of patients. The observed accuracy of MR coronary angiography for detection of hemodynamically significant coronary arterial stenosis is promising, but it needs to be higher before this modality can be used reliably in a clinical setting. PMID- 11012457 TI - Coronary artery bypass grafts: improved electron-beam tomography by prolonging breath holds with preoxygenation. AB - In 45 patients with coronary bypass grafts, the breath-hold interval with and that without preoxygenation was measured. Its effect on depiction of the distal graft anastomosis at electron-beam tomography was evaluated. Preoxygenation prolonged the breath-hold interval in most patients, thereby allowing greater anatomic coverage including more distal anastomoses. Preoxygenation may improve scanning of coronary bypass grafts and increase detectability of graft stenoses. PMID- 11012458 TI - Contrast-enhanced 3D MR angiography with simultaneous acquisition of spatial harmonics: A pilot study. AB - A partially parallel image acquisition technique, simultaneous acquisition of spatial harmonics, or SMASH, was used to increase the spatial and/or temporal resolution in contrast material-enhanced three-dimensional magnetic resonance angiography of the abdominal aorta and renal arteries. In eight healthy subjects, the breath-hold duration was halved at constant spatial resolution, or the spatial resolution was doubled at fixed breath-hold duration, with a 30%-55% reduction in the signal-to-noise ratio but otherwise preserved or improved image quality. PMID- 11012459 TI - Real-time projection MR angiography: feasibility study. AB - Intraarterial injections of small doses of gadopentetate dimeglumine were combined with a fast spoiled-gradient-echo magnetic resonance (MR) sequence to obtain real-time projection angiographic images of the rabbit aorta and canine coronary arteries. Arterial filling and washout, as well as venous and perfusion phases, were clearly displayed, demonstrating that arterial fluoroscopy in which an MR technique is used is feasible. PMID- 11012460 TI - Proton MR spectroscopic imaging without water suppression. AB - To improve reproducibility in proton magnetic resonance (MR) spectroscopic imaging in human brain, simultaneous acquisition of the internal water reference and metabolite signals was evaluated. Measurements in healthy volunteers showed that the increase in dynamic range from signal oversampling was sufficient to avoid digitization errors. In addition, use of singular value decomposition techniques and finite impulse response filters proved effective in separating water and metabolite signals and providing estimates of the metabolite concentrations. PMID- 11012461 TI - MR imaging of acute cervical spine injuries. PMID- 11012462 TI - Academic degrees in the United States and abroad. PMID- 11012464 TI - In memoriam PMID- 11012463 TI - Follow-up of probably benign breast lesions. PMID- 11012465 TI - In memoriam PMID- 11012466 TI - Abstracts of current literature PMID- 11012467 TI - Review article: gastrin and colorectal cancer. AB - The polypeptide hormone gastrin was identified nearly a hundred years ago and its role in the regulation of acid secretion is well established. Gastrin also acts as a growth factor and is trophic for the normal gastric oxyntic mucosa. This growth promoting action has led to the extensive investigation of its role in carcinogenesis, in particular colorectal neoplasia. The relationship between gastrin and colorectal adenocarcinoma has been subject to controversy, however the findings from several recent studies have resulted in a clearer understanding of the mechanism of action of gastrin in this is common cancer. The majority of colorectal cancers produce their own gastrin, which may act in an autocrine manner. The tumour cells also express gastrin/CCKB receptors (and/or a combination of isoforms) which mediate the proliferative action. This locally produced gastrin gives rise to a small increase in systemic gastrin levels. Autocrine gastrin may also have a role in tumour development, as expression occurs early in the adenoma-carcinoma sequence. In addition, several studies using animal models have shown that systemic hypergastrinaemia promotes the proliferation of both normal and neoplastic colonic epithelium. Hyperproliferative colonic epithelium in the presence of hypergastrinaemia has been recorded in humans and a well-designed epidemiological study has demonstrated an increased incidence of colorectal cancer. Gastrin is a potential therapeutic target in the treatment of colorectal cancer and several approaches have been assessed. Receptor antagonists and antisecretory agents have been demonstrated to be ineffectual. Novel methods of inhibition, including the use of anti-gastrin antibodies, are currently being evaluated. PMID- 11012468 TI - Esomeprazole improves healing and symptom resolution as compared with omeprazole in reflux oesophagitis patients: a randomized controlled trial. The Esomeprazole Study Investigators. AB - BACKGROUND: The pharmacologic profile of the new proton pump inhibitor esomeprazole has demonstrated advantages over omeprazole that suggest clinical benefits for patients with acid-related disease. METHODS: 1960 patients with endoscopy-confirmed reflux oesophagitis (RO) were randomized to once daily esomeprazole 40 mg (n=654) or 20 mg (n=656), or omeprazole 20 mg (n=650), the standard recommended dose for RO, for up to 8 weeks in a US, multicentre, double blind trial. The primary efficacy variable was the proportion of patients healed at week 8. Secondary variables included healing and heartburn resolution at week 4, time to first resolution and sustained resolution of heartburn, and per cent of heartburn-free days and nights. Safety and tolerability were also evaluated. RESULTS: Significantly more patients were healed at week 8 with esomeprazole 40 mg (94.1%) and 20 mg (89.9%) vs. omeprazole 20 mg (86.9%), using cumulative life table estimates, ITT analysis (each P < 0.05). Esomeprazole 40 mg was also significantly more effective than omeprazole for healing at week 4 and for all secondary variables evaluating heartburn resolution. The most common adverse events in all treatment groups were headache, abdominal pain and diarrhoea. CONCLUSION: Esomeprazole was more effective than omeprazole in healing and symptom resolution in GERD patients with reflux oesophagitis, and had a tolerability profile comparable to that of omeprazole. PMID- 11012469 TI - CYP2C19 genotype status and intragastric pH during dosing with lansoprazole or rabeprazole. AB - BACKGROUND: CYP2C19 has an important role in the catabolism of several proton pump inhibitors. However, the relative contribution of CYP2C19-mediated metabolism varies among the different proton pump inhibitors. AIM: To determine the effect of CYP2C19 genotype status on intragastric pH during dosing with lansoprazole or rabeprazole. SUBJECTS AND METHODS: The subjects were 20 male volunteers without Helicobacter pylori infection. Their CYP2C19 genotype status was determined by a polymerase chain reaction-restriction fragment length polymorphism method. Twenty-four-hour monitoring of intragastric acidity was performed three times: once without medication, once on the last day of a 7-day course of rabeprazole, and once on the last day of a 7-day course of lansoprazole. RESULTS: Subjects were divided into three groups on the basis of their CYP2C19 genotype status: homozygous extensive metabolizers (homo-EMs, n=7), heterozygous extensive metabolizers (hetero-EMs, n=9), and poor metabolizers (PMs, n=4). The median pH during rabeprazole administration was not influenced by CYP2C19 genotype. On the other hand, the median pH in PMs during lansoprazole dosing was higher than in homo-EMs and hetero-EMs. The percentage of time with pH < 4.0 had a similar tendency to that of median pH. CONCLUSION: CYP2C19 genotype status influences gastric acid suppression by lansoprazole, but not by rabeprazole. PMID- 11012470 TI - Proton pump inhibitors: better acid suppression when taken before a meal than without a meal. AB - BACKGROUND: Proton pump inhibitors including omeprazole and lansoprazole inhibit gastric acid secretion by selectively and non-competitively inactivating the H+, K+ ATPase molecules of the parietal cell, but possibly only those that are actively secreting acid. This might imply that stimulation of acid secretion by a meal is necessary for optimal inhibition of gastric secretion. AIM: To quantify and compare the effect on daytime gastric acidity of omeprazole 20 mg or lansoprazole 30 mg daily taken 15 min before breakfast, with that of the same drug taken without a meal. METHODS: Twenty-one healthy volunteers were randomized to receive either omeprazole or lansoprazole. They were given the drug for two separate periods of 7 days in randomized order and at least 7 days apart. During one period the study medication was taken before breakfast; during the other it was taken at the same hour, but with no meal until 12:00 hours. Lunch was standardized. On day 7, intragastric pH-metry was performed, starting at 08:00 hours. Tracings were analysed for the 8-h period from 08:00 hours until 16:00 hours with regard to percentage time for which gastric pH was below 4.0 and 3.0, and median gastric pH. Tracings were also analysed after removing the 1 h breakfast period, to exclude the buffering effect of the meal. RESULTS: When taking the drug with breakfast, the median percentage time for which gastric pH < 4.0 was 17.2 (interquartile range 4.6-45.5), compared with 42.0 (interquartile range 31.4-48.8) when taken without food (P=0.01). Fifteen subjects had better control of gastric acidity when the medication was taken with breakfast. A pH threshold of 3 and median pH showed similar differences. When the breakfast period was removed, the differences were no longer statistically significant. CONCLUSIONS: When therapy with omeprazole or lansoprazole is indicated, medication should be taken before a meal for optimal control of daytime gastric acidity. PMID- 11012471 TI - A randomized trial of polyurethane and silicone percutaneous endoscopic gastrostomy catheters. AB - BACKGROUND: No data are available on differences in complication rate and long term functioning between polyurethane and silicone percutaneous endoscopic gastrostomy (PEG) catheters. METHODS: We randomized patients who qualified for PEG placement to receive either a polyurethane or silicone PEG catheter. Patients were prospectively monitored for 28 days after placement for the occurrence of complications. Data on long-term PEG survival were obtained retrospectively from the Hospital and general practitioner's medical records. RESULTS: One hundred and six patients were randomized (polyurethane 50, silicone 56). During the first four weeks of follow-up, major complications occurred twice with both polyurethane and silicone PEGs (relative risk 1.1, 95% confidence interval: 0.11 11). Overall complications occurred four times with polyurethane and 17 times with silicone PEGs (relative risk 3.8, 95% confidence interval: 1.37-10.5). Long term follow-up was available in 96 patients. Seven polyurethane PEGs and 10 silicone PEGs were removed because of PEG malfunctioning, the remainder functioned well until death or the reinstitution of oral feeding. The median complication-free survival was 916 days for the polyurethane PEG and 354 days for the silicone PEG (Log rank test: P=0.24). CONCLUSION: Polyurethane PEG catheters were associated with less short-term complications than silicone catheters, but major complications and long-term function were comparable. PMID- 11012473 TI - Abnormal contractile properties of rectal smooth muscle in chronic ulcerative colitis. AB - BACKGROUND: Patients with ulcerative colitis have abnormal rectal motility. AIM: To compare the contractile properties of rectal smooth muscle from patients with ulcerative colitis and controls. METHODS: Rectal smooth muscle strips from patients undergoing resection for ulcerative colitis or cancer (control) were mounted in an organ bath. The effects of carbachol (receptor-mediated) and potassium (causes membrane depolarization) were studied. Acetylcholinesterase histochemistry was performed and nerve counts compared. RESULTS: Ulcerative colitis (n=41) and control (n=34) strips contracted in response to potassium and carbachol. Mean (S.E. M.) maximum response to potassium in the control and ulcerative colitis groups was 1.07 (0.06) g/mg and 1.02 (0.09) g/mg tissue, respectively (P=N.S.). EC50s (concentrations required to give 50% of maximal response) were 75 (1) mM and 73 (1) mM, respectively (P=N.S. ). Although maximum responses to carbachol were similar, 2.12 (0.12) g/mg and 1.95 (0.12) g/mg tissue (P=N.S.), ulcerative colitis strips exhibited an increased sensitivity to carbachol, EC50s: 5.05 x 10-6 (0.55 x 10-6) M vs. 8.36 x 10-6 (0.88 x 10-6) M, P=0.002). There was no significant difference in nerve counts between the tissues, as assessed by staining for acetylcholinesterase. CONCLUSIONS: Ulcerative colitis tissue has an increased sensitivity to carbachol and this is not due to denervation; it may result from increased calcium release from intracellular stores since contraction due to membrane depolarization is not altered. Modulation of this pathway could potentially be used to alter rectal motility in patients with ulcerative colitis. PMID- 11012472 TI - Impaired contractile response of mesenteric arteries in Crohn's disease. AB - BACKGROUND: Crohn's disease is associated with vascular injury and dysregulation of the intestinal immune system which together can provide disturbance of mesenteric circulation functional properties. AIM: To evaluate the vascular reactivity of mesenteric arteries from patients with Crohn's disease. METHODS: Phenylephrine-induced contractions were assessed from 10 patients with Crohn's disease and 8 control organ donors. NG-nitro-L-arginine-methyl-ester (L-NAME) was used to test the presence of inducible NO synthase. Endothelium dependent and independent relaxation was assessed using acetylcholine, bradykinin, calcium ionophore A23187 and sodium nitroprusside. RESULTS: The contractile response to phenylephrine was significantly decreased in arteries without endothelium from patients with Crohn's disease. Exposure to the NO synthase inhibitor L-NAME restored the contractile response to phenylephrine. Relaxation remained unaltered in both groups. CONCLUSION: These data provide direct evidence for fading of contraction caused by phenylephrine in Crohn's disease. The restored mesenteric artery tone by a specific NO synthase inhibitor suggests that an increased production for NO in vascular smooth muscle might be responsible of this altered vascular reactivity. PMID- 11012474 TI - Low serum retinol levels are associated with hepatocellular carcinoma in patients with chronic liver disease. AB - BACKGROUND: Retinol and other vitamin A derivatives affect the differentiation and growth of many tissues and have anti-tumour properties. AIM: To investigate serum retinol levels in patients with liver disease and hepatocellular carcinoma (HCC) and to assess its importance as a risk factor for the development of HCC. METHODS: Serum retinol levels were measured in healthy volunteers and 175 patients (34 with chronic hepatitis C, 117 with cirrhosis, and 24 with HCC. RESULTS: The serum retinol levels (mean +/- s.e.) in ng/mL, were 972.1 +/- 37.7 in the control group and 647 +/- 41.1 in patients with chronic hepatitis C. Serum retinol levels in patients with cirrhosis and HCC were lower than in patients with cirrhosis alone (365.8 +/- 43.1 vs. 438.9 +/- 22.1, P < 0.04). In particular, there was a more significant difference in serum retinol levels between Child-Pugh grade A patients with cirrhosis and Child-Pugh grade A patients with cirrhosis/HCC (serum retinol levels 532.4 +/- 26.7 vs. 366.1 +/- 86.4, P < 0.03). There was a significant difference in serum retinol levels between normal controls and all patients' groups (P < 0.001). There were significantly lower serum retinol levels in cholestatic Child-Pugh grade A patients with cirrhosis compared with noncholestatic Child-Pugh grade A patients with cirrhosis/HCC (411.5 +/- 30.3 vs. 579.7 +/- 32.7, P < 0.0004). Sixty percent of patients with Child-Pugh grade A cirrhosis/HCC had serum retinol levels below 350 ng/mL compared with only 18.4% of cirrhotics without HCC (chi 2-test, P=0.01). No correlation was found between serum retinol levels and alpha FP or any other liver function tests, apart from serum albumin, which showed a positive correlation (r=0.61 P < 0.018). CONCLUSIONS: There was a progressive reduction in serum retinol levels from controls to patients with liver cirrhosis. Those patients with cirrhosis and HCC had significantly lower values than patients with cirrhosis alone. Serum retinol levels may be a risk factor for the development of HCC. PMID- 11012475 TI - Ascorbic acid and intestinal metaplasia in the stomach: a prospective, randomized study. AB - BACKGROUND: Intestinal type metaplasia plays a role in intestinal type gastric carcinoma development. Ascorbic acid demonstrates a protective effect against gastric carcinogenesis, due to its ability to inactivate oxygen free-radicals as well as its nitrite-scavenging effects. AIM: To assess whether long-term ascorbic acid administration following Helicobacter pylori eradication could affect intestinal metaplasia regression in the stomach. METHODS: Sixty-five patients were included in the study. The inclusion criterion was the presence of intestinal metaplasia on the gastric mucosa after H. pylori eradication. An upper gastrointestinal endoscopy was performed and 3 biopsy specimens were taken in the antrum, 3 in the gastric body, and 2 in the incisura angularis. Patients were randomized to receive 500 mg of ascorbic acid o.d., after lunch (32 patients) for 6 months or no treatment (33 patients). All patients underwent to endoscopic control at the end of the 6 months. RESULTS: H. pylori infection recurrence was detected in 6 (9.4%) patients (three from each group), and these patients were excluded from further analysis. We were unable to find evidence of intestinal metaplasia in any biopsied site of the gastric mucosa in 9/29 (31%) patients from the ascorbic acid group and in 1/29 (3.4%) of the patients from the control group (P=0.006). Moreover, a further six (20.7%) patients from the ascorbic acid group presenting chronic inactive pangastritis with widespread intestinal metaplasia at entry, showed less extensive antritis with intestinal metaplasia at control, whilst a similar finding was only seen in one patient from the control group (P=0.051). CONCLUSION: The administration of ascorbic acid significantly helps to resolve intestinal metaplasia of the gastric mucosa following H. pylori eradication, and its use as a chemoprevention treatment should be considered. PMID- 11012476 TI - Helicobacter pylori-gastrin link in MALT lymphoma. AB - BACKGROUND: There is accumulating evidence for the role of Helicobacter pylori in the development of gastric cancer as well as of lymphomas that arise in mucosa associated lymphoid tissue (MALT). We reported recently that gastric cancer patients show high prevalence of cagA-positive H. pylori and express gastrin and gastrin receptors enabling them to stimulate tumour growth in autocrine fashion. AIMS: Since the H. pylori infection is considered to be more strongly associated with MALT lymphoma than with gastric cancer, we decided to determine the gastrin and its receptors' mRNA expression and gastrin content in this tumour as well as the release of this hormone both into plasma and gastric lumen. Twenty MALT lymphoma patients were compared with 100 age- and gender-matched controls with similar dyspeptic symptoms. RESULTS: The overall H. pylori seropositivity in MALT lymphoma was about 90% and CagA positivity was 70%, compared to 56% and 33%, respectively, in controls. The serum gastrin in MALT lymphoma was about sixfold higher than in controls while gastric luminal gastrin in these patients was over 70 times higher than in controls. Gastrin content in tumour was about 10-fold higher than in antral mucosa. Gastrin and gastrin-receptor (CCKB-receptor) mRNA were detected by reverse transcriptase-polymerase chain reaction in cancer tissue whilst in the fundic and antral mucosa, only enhanced expression of CCKB-receptor mRNA and gastrin mRNA was detected, respectively. Histamine stimulation in MALT lymphoma induced acid secretion that was only about 30% of control value due to atrophic gastritis. This study confirms an important role of CagA-positive H. pylori in the pathogenesis of MALT lymphoma and shows that this lymphoma is capable of synthesizing and releasing potent growth promoting gastrin, possibly due to the action on G-cells of H. pylori-originated Nalpha-methyl histamine and cytokines (tumour necrosis factor alpha and interleukin-8). CONCLUSIONS: Gastric MALT lymphoma is closely linked to CagA-positive H. pylori infection. Gastrin and its receptors may be implicated in the pathogenesis of gastric lymphoma. PMID- 11012477 TI - Proton pump inhibitor, clarithromycin and either amoxycillin or nitroimidazole: a meta-analysis of eradication of Helicobacter pylori. AB - AIM: To perform a meta-analysis of studies comparing twice daily, one-week triple therapy with a proton pump inhibitor, clarithromycin (C) and amoxycillin (A) (PCA) vs. those using proton pump inhibitor, clarithromycin and a nitroimidazole (N) (PCN) for H. pylori eradication. REVIEW METHODS: SELECTION CRITERIA: Comparative randomized trials of PCA vs. PCN were included. DATA SOURCES: PubMed database and abstracts from congresses until September 1999. STATISTICS: Meta analysis was performed combining the Odds Ratios (OR) of the individual studies in a global OR (Peto method) both on an intention-to-treat (ITT) and on a per protocol (PP) basis. RESULTS: Twenty-two studies fulfilled the inclusion criteria. Eighteen studies reported ITT and 20 PP analysis. Mean H. pylori eradication rates were 81% (95% CI: 79-83%) ITT, and 84% (82-86%) PP with PCA, and 81% (78-83%) ITT and 84% (82-86%) PP with PCN; the odds ratio for the effect of PCA vs. PCN was 1 (0.83-1.22) on an ITT, and 0.98 (0.8-1.2) on a PP basis. Subanalysis showed that mean H. pylori eradication efficacy with PC(250 b.d.)A was 81% (78-85%) ITT, vs. 86% (83-89%) with PC(250 b.d.)N. The odds ratio for this comparison was 0.68 (0.48-0.98). Finally, when comparing PC(500 b.d. )A against PC(250 b.d.)N ITT cure rates were 77% (74-80%), and 75% (72-78%) with an odds ratio of 1.18 (0.93-1.5). CONCLUSION: Overall, one-week combination regimens of PCA and PCN present similar H. pylori eradication efficacy. Nevertheless, the PCN regimen obtains significantly better results when using low doses of C (250 mg b.d.). PMID- 11012478 TI - Helicobacter pylori-positive duodenal ulcer: three-day antibiotic eradication regimen. AB - BACKGROUND: The most widely used treatments for ulcer healing and Helicobacter pylori eradication consist of a 1-2 week regimen of a proton pump inhibitor plus two or three antimicrobials. AIMS: To evaluate the efficacy, safety, cost, and tolerance of a three-day regimen with three antibiotics vs. a 10-day treatment with a proton pump inhibitor or vs. a ranitidine bismuth citrate triple therapy. METHODS: Two hundred and twenty-one patients with endoscopically-proven H. pylori positive duodenal ulcers were recruited to the study. Recruited patients were assigned to one of the following four regimens: (I) omeprazole 40 mg o.m. plus amoxycillin 1 g b.d. and clarithromycin 500 mg b.d. for 10 days (OAC: 55 patients); (ii) omeprazole 40 mg o.m. on days 1-5, plus amoxycillin 1 g b.d., clarithromycin 500 mg b.d. and metronidazole 500 mg b.d. on days 3-5 (OACM: 56 patients); (iii) ranitidine bismuth citrate 400 mg b.d. plus amoxycillin 1 g b.d. and clarithromycin 500 mg b.d. for 10 days (RAC: 54 patients); (iv) ranitidine bismuth citrate 400 mg b.d. on days 1-5, plus amoxycillin 1 g b.d., clarithromycin 500 mg b.d. and metronidazole 500 mg b.d. on days 3-5 (RACM: 56 patients). Fisher's exact test was used to compare data regarding healing and eradication in the four groups. RESULTS: The intention-to-treat eradication and ulcer healing rates for the RACM regimen were 95% and 98%, respectively. Statistically significant differences were observed, relating to the eradication and healing of ulcers, between RACM and either the RAC or OAC regimens. CONCLUSION: The three-day antibiotic therapy with amoxycillin, clarithromycin and metronidazole in addition to ranitidine bismuth citrate is a very effective anti H. pylori regimen. PMID- 11012479 TI - Third line treatment for Helicobacter pylori: a prospective, culture-guided study in peptic ulcer patients. AB - BACKGROUND: A third line treatment is needed in roughly 5% of patients infected with Helicobacter pylori. Few data have been reported on efficacy of treatment regimens in these patients. METHODS: A prospective trial was designed to study the effectiveness of third line treatment of H. pylori infection in ulcer patients. Two-week quadruple, culture-guided, combinations were used in 31 consecutive patients. Susceptibility to metronidazole and clarithromycin were studied by E-test, and thereafter a predetermined treatment regimen was used. Compliance was evaluated by pill count, and eradication defined by negative urea breath test at 6 weeks. RESULTS: Two main quadruple regimens were used in 29 patients. In spite of good compliance, the combination of omeprazole, tetracycline, bismuth and clarithromycin (OTBC) showed an eradication rate (per protocol analysis) of 36% (five out of 14; CI: 12.8-64.9), and if amoxycillin was used (OTBA) the rate was 67% (eight out of 12; CI: 34.9-90.1). The difference was not significant. No clinical factor was found to be associated with failure to eradicate. CONCLUSIONS: Third line treatment often fails to eradicate H. pylori infection. New strategies need to be developed and tested for this common clinical situation. PMID- 11012480 TI - Efficacy of two one-week rabeprazole/levofloxacin-based triple therapies for Helicobacter pylori infection. AB - BACKGROUND: One-week low-dose proton pump inhibitor-based triple therapies have usually proved to be effective treatments for Helicobacter pylori infection. AIM: To investigate the eradication efficacy, safety profile and patient compliance of two triple therapies containing a standard dose of rabeprazole and a new fluoroquinolone, levofloxacin. METHODS: One hundred patients referred to us for gastroscopy, who were H. pylori-positive, were consecutively recruited in a prospective, open-label study. The enrolled patients were randomised to receive a seven-day course of rabeprazole 20 mg o.d. plus levofloxacin 500 mg o.d. and either amoxycillin 1 g b.d. (RLA group) or tinidazole 500 mg b.d. (RLT group). Their H. pylori status was assessed by means of histology and rapid urease test at entry, and by 13C-urea breath test 8 weeks after the end of treatment. RESULTS: All 100 enrolled patients completed the study. Forty-six of 50 patients treated with RLA (both PP and ITT analysis: 92%; 95% CI: 81-98%) and 45 of 50 with RLT (both PP and ITT analysis: 90%: 95% CI: 78-97%), became H. pylori negative. Slight or mild side-effects occurred in 4 (8%) patients of the RLA group and in 5 (10%) of the RLT group. CONCLUSIONS: This study demonstrates the efficacy of two 1-week rabeprazole-based triple therapies including levofloxacin to eradicate H. pylori. These regimens prove to be safe, well-tolerated, and achieved good eradication rates. Levofloxacin may be an effective alternative to clarithromycin in triple therapy regimens. PMID- 11012481 TI - Changes in Helicobacter pylori-induced gastritis in the antrum and corpus during long-term acid-suppressive treatment in Japan. AB - BACKGROUND: Several studies have shown that acid-suppressive therapy aggravates corpus gastritis in patients with Helicobacter pylori infection, promoting the development of atrophic gastritis. AIM: To study the effects of long-term use of antisecretory agents on the H. pylori-positive gastric mucosa in Japan, a country with a high incidence of gastric cancer. METHODS: A total of 141 H. pylori positive patients who had peptic ulcers or reflux oesophagitis were treated for 3 years with either omeprazole (20 mg/day) alone (n=7) or with omeprazole for primary therapy (8 weeks), followed by famotidine (40 mg/day) for maintenance therapy (n=134). Endoscopy was performed before, during, and after treatment. Biopsy specimens were taken from the greater curvature of the antrum and corpus and were examined histologically. RESULTS: The long-term use of famotidine after 8 weeks of treatment with omeprazole distinctly decreased H. pylori density and neutrophil infiltration in the antrum, but did not change H. pylori density in the corpus. The gastritis score increased in patients who had no, or only mild corpus gastritis before treatment (n=74), and significantly decreased in those who had moderate or severe gastritis before treatment (n=60). In four of the seven patients who received long-term treatment with omeprazole alone, neutrophil infiltration and H. pylori density decreased not only in the antrum but also in the corpus. There was no increase in intestinal metaplasia or mucosal atrophy as assessed endoscopically during follow-up. CONCLUSION: Changes in corpus gastritis in response to acid-suppressive therapy depend on the severity of gastritis before treatment. Long-term use of acid-suppressive therapy apparently does not accelerate the development of atrophy or intestinal metaplasia in Japanese patients. PMID- 11012482 TI - (13)C-urea breath test without a test meal is highly accurate for the detection of Helicobacter pylori infection in Chinese. AB - BACKGROUND: Conventional (13)C-urea breath testing ((13)C-UBT) includes a test meal to delay gastric emptying, which, theoretically, improves the accuracy of the test. Citric acid has been proposed as the best test meal. However, recent studies have suggested that a test meal may not be necessary. AIM: To investigate a new (13)C-UBT protocol without a test meal in a Chinese population. METHODS: Consecutive dyspeptic patients referred for upper endoscopy were recruited. (13)C UBT was performed on two separate days with or without a test meal (2.4 Gm citric acid) and compared with the 'gold standard' (CLO test and histology). RESULTS: Two hundred and two patients were tested. Using receiver operating characteristics (ROC) analysis, the optimal delta-value and optimal measurement interval for UBT were 5% and 30 min, respectively, both with or without a test meal. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of (13)C-UBT with citric acid (96.5%, 97.7%, 98.2%, 95.6%, 97.0%) were similar to (13)C-UBT without a test meal (94.7%, 97.7%, 98.2%, 93.5%, 96.0%). CONCLUSION: This simplified (13)C-UBT protocol without a test meal produced highly accurate and reliable results in the Chinese population. PMID- 11012483 TI - Lower-dose (13)C-urea breath test to detect Helicobacter pylori infection comparison between infrared spectrometer and mass spectrometry analysis. AB - BACKGROUND: The expense of the (13)C-urea breath test (UBT) to detect Helicobacter pylori infection is mainly due to the cost of (13)C-urea and the analysis using isotope ratio mass spectrometry (IRMS). AIM: To test whether a UBT, using a lower dose of urea and lower-priced isotope-selective nondispersive infrared spectrometry (INIS), can preserve diagnostic efficacy in clinical practice. METHODS: A total of 177 dyspeptic patients received endoscopy for H. pylori culture and histology. All of them received a UBT in which the duplicate baseline, 10 min, and 15 min breath samples after ingestion of 50 mg (13)C-urea were collected to analyse the excess (13)CO(2)/(12)CO(2) ratio (ECR) by IRMS (ABCA, Europa Scientific, UK) and INIS (UBiT-IR200, Photal Otsuka Electronics, Japan), respectively. RESULTS: Of the 177 patients, 84 were infected and 93 were uninfected with H. pylori. A close correlation of ECR was found between IRMS and INIS (r=0.9829 at 10 min; r=0.9918 at 15 min, P < 0.0001). Analysing the 15-min samples, UBT by both IRMS and INIS achieved the same sensitivity (96. 4%) and specificity (98.9%). CONCLUSIONS: INIS is as effective as IRMS for UBT, and can use a lower dose of (13)C-urea. This can provide an economic UBT, using the lower priced INIS and a low dose of (13)C-urea. PMID- 11012484 TI - Gastric acid secretion in cyclooxygenase-1 deficient mice. AB - BACKGROUND: Constitutive cyclooxygenase-1 enzyme synthesizes prostaglandins which are thought to play an important role in the functional integrity of the stomach gastric mucosa. Recently, it was shown that cyclooxygenase-1 deficient mutant mice did not develop spontaneous gastric pathology and appear less sensitive to indomethacin-induced gastric damage. AIM: To investigate gastric acid secretion in cyclooxygenase-1 deficient mutant mice. METHODS: The basal and histamine or isobutyl methylxanthine-stimulated acid secretion in stomachs of cyclooxygenase-1 deficient homozygous mice and the effect of indomethacin was compared with that of heterozygous and wild-type mice using isolated lumen perfused mouse stomachs, in organ baths, monitored by pH-electrodes. RESULTS: There was no significant difference in the basal or histamine stimulated gastric acid secretion between wild-type or heterozygous or homozygous mice. However, isobutyl methylxanthine was more potent in the cyclooxygenase-1 deficient and heterozygous mice than in wild-type mice. Indomethacin, at concentrations below 1 mM, had no effect on either basal or histamine stimulated acid secretion in any of the mice populations. CONCLUSION: Gastric acid secretion is maintained without prostaglandin involvement in cyclooxygenase-1 deficient mice. The finding that basal and histamine-stimulated gastric acid secretion was similar in the cyclooxygenase-1 deficient, compared to wild-type mice is consistent with the lack of spontaneous gastric pathology in the cyclooxygenase-1 deficient mice. PMID- 11012485 TI - Local and systemic effects of peritoneal lavage with high concentrations of adenosine in rats. AB - BACKGROUND: Adenosine exerts actions which may be beneficial in treating diseases of the gastrointestinal tract. However, administered systemically, adenosine causes a 'stress reaction' and may adversely affect blood pressure and cardiac and renal function. AIM: To determine whether peritoneal lavage with adenosine provides pharmacological levels of adenosine in the intestines without elevating adenosine levels in the systemic circulation. METHODS: Rats received an intramesenteric artery infusion of angiotensin II (30 ng/min) plus methoxamine (3 microg/min) to reduce mesenteric blood flow by approximately 60%, and adenosine solutions were instilled into the abdominal cavity. In a second study, microdialysis probes were placed in the mesenteric vein and aortic arch of rats, and the peritoneal cavity was continuously lavaged with adenosine solutions. RESULTS: High concentrations (10(-3)M) of adenosine normalized the mesenteric blood flow without affecting blood pressure or heart rate. High concentrations of adenosine (10(-3)M) induced micromolar levels of adenosine and inosine in the mesenteric vein, without affecting adenosine or inosine levels in the aorta. CONCLUSIONS: Peritoneal lavage with high concentrations of adenosine provides pharmacological levels of adenosine in the gastrointestinal tract without systemic side-effects. Peritoneal lavage with high concentrations of adenosine may be useful for the treatment of a number of diseases of the gastrointestinal tract. PMID- 11012486 TI - Anaesthesia, what's in a name? - time for change. PMID- 11012487 TI - Substance misuse amongst anaesthetists. PMID- 11012488 TI - Substance misuse amongst anaesthetists in the United Kingdom and Ireland. The results of a study commissioned by the Association of Anaesthetists of Great Britain and Ireland. AB - Three hundred and four departments of anaesthesia in UK and Ireland were sent questionnaires about alcohol and drug abuse in anaesthetists over the preceding 10-year period. Information was sought on the nature and extent of substance problems, their presentation and management. The survey achieved a high response rate of 71.7% and a total of 130 cases were reported, of whom 34.6% were consultants and 43.2% were trainees. Over 50% of respondents felt a lack of confidence in dealing with alcohol or drug misuse amongst colleagues. The results of this survey demonstrate that over one anaesthetist per month has presented with significant alcohol or drug misuse in the UK and Ireland over the last 10 years. It is important that those with management responsibilities for departments of anaesthesia are aware that such problems exist and are likely to impact on the professional ability and health of the affected individual. The Working Party on Substance Abuse at the Association of Anaesthetists has recently published guidance in the management of these problems. A case is made for increasing awareness in this sensitive subject to enable early recognition and treatment of an anaesthetist who is misusing alcohol and drugs since intervention can be effective. PMID- 11012489 TI - Closed loop control of anaesthesia: an assessment of the bispectral index as the target of control. AB - We investigated the performance of a closed-loop system for administration of general anaesthesia, using the bispectral index as a target for control. One hundred patients undergoing gynaecological or general surgery were studied. In 60 patients, anaesthesia was maintained by intravenous infusion of a propofol/alfentanil mixture. In 40, an isoflurane/nitrous oxide based technique was used. For each technique, patients were randomly allocated to receive either closed-loop or manually controlled administration of the relevant agents (propofol/alfentanil or isoflurane), with an intra-operative target bispectral index of 50 in all cases. Closed-loop and manually controlled administration of anaesthesia resulted in similar intra-operative conditions and initial recovery characteristics. During maintenance of anaesthesia, cardiovascular and electro encephalographic variables did not differ between closed-loop and manual control groups and deviation of bispectral index from the target value was similar. Intra operative concentrations of propofol, alfentanil and isoflurane were within normal clinical ranges. Episodes of light anaesthesia were more common in the closed-loop group for patients receiving propofol/alfentanil anaesthesia and in the manual group for patients receiving isoflurane/nitrous oxide anaesthesia. Convenience aside, the closed-loop system showed no clinical advantage over conventional, manually adjusted techniques of anaesthetic administration. PMID- 11012490 TI - A comparison of antagonism of rocuronium-induced neuromuscular blockade during sevoflurane and isoflurane anaesthesia. AB - Volatile anaesthetic agents potentiate neuromuscular blocking agents and retard their rate of reversal. We hypothesised that there was a difference in the rate of reversal of rocuronium-induced neuromuscular blockade based on the selection of inhalation agent. Thirty-eight patients undergoing elective surgical procedures received either sevoflurane or isoflurane, by random allocation. Neuromuscular blockade was induced using rocuronium 0.6 mg.kg-1 followed by continuous intravenous infusion to maintain 90% suppression of the single twitch response. Upon completion of surgery, the rocuronium infusion was discontinued, neostigmine 50 microg.kg-1 and glycopyrrolate 10 microg.kg-1 were administered. Times from reversal to T1 = 25, 50 and 60% and train-of-four ratio = 0.6 were recorded. The mean (SD) times to train-of-four ratio = 0.6 in the isoflurane and sevoflurane groups were 327 (132) and 351 (127) s, respectively. The mean (SD) times to single twitch response T1 = 25, 50 and 60% in the isoflurane group were 81 (33), 161 (59) and 245 (84) s, respectively, and in the sevoflurane group were 95 (35), 203 (88) and 252 (127) s, respectively. It is concluded that reversal of rocuronium-induced neuromuscular blockade is similar during isoflurane and sevoflurane anaesthesia. PMID- 11012491 TI - Determination of sevoflurane alveolar concentration for tracheal intubation with remifentanil, and no muscle relaxant. AB - The sevoflurane alveolar concentration needed for tracheal intubation with remifentanil was studied in 26 adult patients premedicated with 100 mg hydroxyzine. Anaesthesia was induced with sevoflurane in oxygen. The concentration was determined by Dixon's up-and-down method. The first patient was tested at 4.5%. One minute after obtaining the preselected value, remifentanil 1 microg.kg-1 was injected for 60 s followed by an infusion of 0.25 microg.kg-1. min-1. Ventilation was then manually assisted for 2 min and tracheal intubation was attempted. Tracheal intubation conditions and responses to intubation were noted. Heart rate and mean arterial pressure were recorded before induction, before remifentanil injection, and before and 3 min after tracheal intubation. The concentration for acceptable intubating conditions was 2.5 +/- 0.7%. From logistic regression, ED50 and ED95 were 2.0% (95% CI 1.3-2.5) and 3.2% (95% CI 2.6-5.6), respectively. With sevoflurane 2.5%, heart rate and mean arterial pressure decreased by 18% and 15%, respectively, after remifentanil administration and increased slightly after tracheal intubation. PMID- 11012492 TI - Effects of peribulbar bupivacaine as an adjunct to general anaesthesia on peri operative outcome following retinal detachment surgery. AB - Sixty premedicated, ASA physical status I or II patients weighing > 25 kg scheduled for elective retinal detachment repair were randomly assigned to receive either peribulbar block with 10 ml of 0.25% bupivacaine (block group) or intravenous morphine 150 microg.kg-1 (morphine group), prior to the induction of general anaesthesia (n = 30 in each group). Patients were evaluated for intra operative oculocardiac reflex, peri-operative pain relief, recovery from anaesthesia and postoperative nausea and vomiting. Apart from significantly reducing the incidence of oculocardiac reflex (30% vs. 70%, p = 0.0019), peribulbar bupivacaine also attenuated the severity of the reflex. Postoperative pain relief was superior in the block group. More block group patients had the maximum recovery score in the immediate postoperative period (80% vs. 27%, p < 0. 0001) and they achieved complete recovery significantly faster than the morphine group (17.3 (14.7) min vs. 66.7 (29.7) min, p < 0.0001). The incidence (40% vs. 77%, p = 0.004) and severity of postoperative nausea and vomiting were significantly less in the block group. In summary, peribulbar bupivacaine, when administered together with general anaesthesia, attenuated oculocardiac reflex, provided comparable intra-operative and superior postoperative analgesia, resulted in significantly earlier and better recovery from anaesthesia, and significantly reduced the incidence and severity of postoperative nausea and vomiting. PMID- 11012493 TI - Protective effects of plasma replacement fluids on erythrocytes exposed to mechanical stress. AB - Haemoglobin release from 40 suspensions of packed red blood cells in modified fluid gelatin, 4% albumin solution, 6% hydroxyethyl starch and normal saline was investigated in vitro during circulation with a roller pump from a heart-lung machine for 120 min at a flow rate of 2.5 l.min-1 at room temperature. The lowest haemoglobin release was obtained with erythrocytes in modified fluid gelatin, whereas free haemoglobin concentrations became progressively higher with albumin, hydroxyethyl starch and normal saline [median free haemoglobin (interquartile range) after 120 min circulation: gelatin 493 (360-601) mg.l-1, albumin 692 (590 1111) mg.l-1, hydroxyethyl starch 1121 (692-1518) mg.l-1, normal saline 1178 (881 1757) mg.l-1, p < 0.001]. Modified fluid gelatin appears to have potent erythrocyte protective properties similar to those of albumin. This effect could decrease mechanical haemolysis during extracorporeal circulation or cell saver autotransfusion if modified fluid gelatin is used as part of a priming solution or as an additive in wash solutions. PMID- 11012494 TI - Peripheral nerve injuries associated with anaesthesia. AB - Peripheral nerve injuries can occur at any time during the peri-operative period. The long-term disability that results may have serious consequences for a patient. The incidence of peri-operative nerve injuries can be reduced by anaesthetists being aware of their causes and pathophysiology. This review article aims to explain the incidence, pathophysiology and medicolegal implications of peri-operative nerve injury and provides suggestions as to how they may best be avoided. PMID- 11012495 TI - Supplementary oxygenation with the laryngeal mask airway: a comparison of four devices. AB - The provision of supplementary oxygen via the laryngeal mask airway used in the recovery room is important for patient safety. Several devices have been described for this purpose, but these studies have not included an accurate measurement of the most clinically important variable, the end-tidal oxygen concentration. We constructed an artificial model of spontaneous ventilation to compare the efficacy and safety of four devices; a circuit filter, a Hudson mask, the 'T-bag' device and a T-piece. We combined the use of oximetry with a pneumotachograph to provide a continuous picture of the oxygen delivery characteristics of the devices at flow rates of 2, 4 and 8 l.min-1. The performances of the T-bag and the T-piece were superior to those of the filter and Hudson mask, with end-tidal oxygen concentrations of 46.1%, 45.8% and 35.4%, 34.8%, respectively, at 8 l.min-1. Single point assessments of oxygen delivery, such as peak inspired oxygen concentration, may overestimate the efficacy of test devices. PMID- 11012496 TI - Drawbacks of pushovers. AB - Some military anaesthetists have started to use the Oxford Miniature Vaporiser in a pushover configuration with the Triservice anaesthetic apparatus. This vaporiser performs identically in the pushover and drawover configurations with the Cape TC 50 ventilator. We tested the Oxford Miniature Vaporiser with three other ventilators and found variable performance. When used in the pushover configuration with the Laerdal bag at normal minute volumes, the Oxford Miniature Vaporiser delivers a higher than set output. PMID- 11012497 TI - A double-blind randomised comparison of ropivacaine 0.5%, bupivacaine 0.375% - lidocaine 1% and ropivacaine 0.5% - lidocaine 1% mixtures for cataract surgery. AB - This study evaluated the efficacy and side-effects of plain ropivacaine compared with ropivacaine-lidocaine and bupivacaine-lidocaine mixtures for peribulbar blocks in cataract surgery. Ninety patients were randomly allocated to three groups and received peribulbar blockade using one of the three solutions. Speed of onset and quality of blockade were assessed using akinesia, surgical satisfaction and patient satisfaction. Complications and cardiovascular side effects were noted. There was a slower onset of akinesia using ropivacaine alone, although at 10 min after injection all groups were equal in this respect. There was no difference in surgical or patient satisfaction between the groups. There were no differences in pain on injection, preblock and postblock blood pressure, heart rate or oxygen saturation. The optimal time to surgical incision after peribulbar blockade is not less than 15 min and plain ropivacaine fulfils this criterion. PMID- 11012498 TI - The effect of the addition of ropivacaine or bupivacaine upon pruritus induced by intrathecal fentanyl in labour. AB - Sixty patients in early labour were randomly allocated to one of three groups. The control group received intrathecal fentanyl 25 microg, the ropivacaine group received intrathecal fentanyl 25 microg and ropivacaine 2.5 mg while the bupivacaine group received intrathecal fentanyl 25 microg and bupivacaine 2.5 mg. The incidence of pruritus was 100% in controls, compared with 85% in the ropivacaine group (not significant) and 75% in the bupivacaine group (p = 0.003). The severity of pruritus was significantly less in the ropivacaine (p = 0.006) and bupivacaine (p = 0.001) groups. Most patients developed pruritus by 30 min. Pruritus above the abdomen was not reduced in patients receiving local anaesthetics. There were no significant differences in the mean pain visual analogue score, systolic blood pressure, maternal heart rate and upper level of reduced pin-prick sensation in the first 30 min. Intrathecal ropivacaine and, more so, intrathecal bupivacaine reduce the incidence and severity of pruritus from intrathecal fentanyl for labour analgesia. PMID- 11012499 TI - Reducing red blood cell transfusion in elective surgical patients: the role of audit and practice guidelines. AB - In 1996, we prospectively audited peri-operative transfusion practice in elective surgical patients over a 3-month period. Two-unit transfusions represented 60% of all transfusions. Haemoglobin was measured infrequently prior to transfusion and the main 'trigger' for transfusion was an estimated blood loss in excess of 500 ml. Transfusion guidelines that required the haemoglobin level to be measured immediately before transfusion were introduced. The audit was repeated in 1998; transfusion 'triggers' and the number of transfusions for the two periods were compared. In the second audit, the total number of transfusions decreased by 43%. The mean estimated blood loss associated with a 2-unit transfusion had increased from 608 (373) ml to 1320 (644) ml (p < 0.01) and the estimated haemoglobin concentration after transfusion had decreased from 12.4 (1.8) g.dl-1 to 9.9 (2.4) g.dl-1 (p < 0.01). These results suggest that transfusion guidelines can have a significant impact on clinical practice. PMID- 11012500 TI - The incidence of transient neurological symptoms after spinal anaesthesia with lidocaine compared to prilocaine. AB - The purpose of this double-blind study was to investigate the incidence of transient neurological symptoms after the use of isobaric lidocaine and isobaric prilocaine for spinal anaesthesia. Seventy patients (ASA 1-2, age between 18 and 70 years) were randomly assigned to two groups of 35 patients each, to receive either isobaric 2% lidocaine 4 ml or isobaric 2% prilocaine 4 ml intrathecally, at the L3-4 interspace. One patient in the prilocaine group could not be included because data were incomplete. On the first postoperative day, patients were evaluated for transient neurological symptoms. Pain was scored on a 10-point scale. Seven patients (20%) in the lidocaine group had transient neurological symptoms with a mean pain score of 5.3, whereas no patient in the prilocaine group had these complaints (p = 0.006). Symptoms disappeared within 4 days. Prilocaine results in a lower incidence of transient neurological symptoms than lidocaine intrathecally and therefore it is more suitable for short surgical procedures. PMID- 11012501 TI - Paediatric intensive care transfers: 2. PMID- 11012502 TI - Paediatric intensive care transfers 3. PMID- 11012503 TI - A further comment PMID- 11012504 TI - Stress and trainees. PMID- 11012505 TI - Testing breathing systems. PMID- 11012506 TI - Fibreoptic intubation through a laryngeal mask airway facilitated by a guide wire. PMID- 11012507 TI - Auditing the application of cricoid pressure. PMID- 11012508 TI - Classification of laryngoscopic view. PMID- 11012509 TI - A novel method of delivering vasoconstrictors to the nasal passage. PMID- 11012510 TI - Failure of a 'Diprivan 1%' prefilled propofol syringe. PMID- 11012512 TI - Tec 6 vaporiser raises the temperature. PMID- 11012513 TI - The effect of smoking on postoperative nausea and vomiting. PMID- 11012515 TI - Cardiac arrest after video-assisted thoracoscopic drainage of pleural effusion. PMID- 11012516 TI - An unexpected difficult extubation. PMID- 11012517 TI - Urgent abdominal aortic aneurysm repair and sickle cell trait. PMID- 11012518 TI - Survival following ruptured aortic aneurysm. PMID- 11012519 TI - Internal jugular cannulation using a long spinal needle as a seeker needle. PMID- 11012520 TI - Bronchoscopic confirmation of the correct placement of the tracheostomy tube during translaryngeal tracheostomy. PMID- 11012522 TI - Transurethral vaporisation of the prostate. PMID- 11012523 TI - Iatrogenic foreign body in the bronchus. PMID- 11012524 TI - Succinylcholine-associated postoperative myalgias...not always! PMID- 11012525 TI - Anaesthesia and angiotensin II receptor antagonist. PMID- 11012526 TI - Remifentanil and seizures. PMID- 11012527 TI - Anaphylaxis caused by neostigmine. PMID- 11012528 TI - Soya bean oil in Diprivan. PMID- 11012529 TI - Vancomycin causes dangerous precipitation when infused with gelatin fluid. PMID- 11012530 TI - Should Mendelson's syndrome be renamed? PMID- 11012531 TI - Identification of the epidural space. PMID- 11012532 TI - Keep epidurals simple. PMID- 11012533 TI - Pitfalls of moisturising creams. PMID- 11012534 TI - Generic propofol: what's the point? PMID- 11012536 TI - Protecting pacemakers in prone patients. PMID- 11012535 TI - Airway protection during percutaneous tracheostomy. PMID- 11012537 TI - Hemolipodialysis attenuates oxidative stress and removes hydrophobic toxins. AB - Uremic patients undergoing hemodialysis often have increased oxidant stress and accumulation of uremic toxins. Hemodialysis, per se, often can exacerbate oxidant stress and may be inefficient at removing hydrophobic or protein bound toxins. We describe a new hemodialytic method that incorporates liposomes and antioxidants to remove hydrophobic/uremic toxins and minimize free radical mediated damage. In vitro experiments measured advanced oxidation protein products (AOPP), malonaldehyde, reactive carbonyls, and the removal of platelet activating factor (PAF) and bilirubin during extracorporeal circulation with or without liposomes. We observed a significant reduction of oxidation products as well as a significant removal of PAF and bilirubin compared to normal hemodialysis. PMID- 11012538 TI - Carotid intima-media thickness and plaques in hemodialysis patients. AB - Atherosclerosis is accelerated in hemodialysis patients. Using B-mode ultrasonography, we compared intima-media thickness (IMT) and the prevalence of plaques in the common carotid and internal carotid arteries in 28 randomly selected hemodialysis patients with that in 28 age- and sex-matched normal controls. The IMT values of the common carotid and internal carotid arteries were higher in hemodialysis patients than in controls with more hemodialysis patients having plaques. In hemodialysis patients, there was a relationship between age and IMT in the common carotid arteries, in the area of bifurcation, and in the internal carotid arteries. We found no relationship between IMT and atherosclerotic risk factors or duration of hemodialysis treatment. IMT at all sites correlated with the number of plaques. Age was the only significant determinant for number of plaques. The results indicate that hemodialysis patients showed advanced atherosclerosis in the carotid arteries compared with age- and sex-matched normal subjects. PMID- 11012540 TI - Plasmapheresis-related hypotension. AB - Hypotension is an uncommon complication of procedures involving extracorporeal circulation, including plasmapheresis. From November 1993 to March 1999, we treated 139 patients who underwent a total of 1,137 sessions of double filtration plasmapheresis (DFP). Hypotension was defined as a systolic blood pressure (BP) < 80 mm Hg or any decrease of systolic BP with systemic reactions. A total of 17 (1.5%) episodes of hypotension were documented in 15 patients during the study period. Hypotensive episodes occurred in 2.3% of patients with inflammatory neuropathy, 1.2% of patients with myasthenia gravis, and 1.2% of patients with all other medical diseases. Involvement of the autonomic nerve system (ANS) and a low baseline BP were associated with the occurrence of hypotension. Eight (47%) of 17 episodes were symptomatic and 2 were complicated with seizure. Patients with symptomatic hypotension had a higher level of systolic BP prior to DFP and a larger drop of systolic BP and pulse rate during hypotensive attacks compared to asymptomatic patients. Most hypotensive episodes were resolved briefly after intravenous infusion of saline within 30 min. Eight (47%) of the hypotensive episodes occurred during the first session of DFP treatment. Twelve (71%) of 17 episodes occurred during the last half period of treatment; 6 of them were noted during the terminating stage of DFP. In conclusion, in this series plasmapheresis related hypotension occurred in 1.5% of DFP sessions and had a higher prevalence in patients with ANS instability and low BP. Extra caution in monitoring BP during DFP therapy is warranted in these vulnerable patients, especially during the termination phase of the first DFP session. PMID- 11012539 TI - Ex vivo biocompatibility of avidin-agarose: a new device for direct adsorption of biotinylated antibodies from human whole blood. AB - Radioimmunotherapy using radiolabeled antitumor antibodies (RAA) is limited by the toxicity of unbound antibodies in the circulation. Removal of excessive antibodies by affinity-adsorption could therefore allow the administration of increased dosages of RAA while decreasing their adverse effects. Recently, avidin agarose (AA) minicolumns were used in animal experiments for the removal of biotinylated antibodies from whole blood exploiting the high affinity binding of biotin to avidin (pK 1015 M-1). This study was performed to evaluate the ex vivo biocompatibility of AA minicolumns with human blood. Ten ml AA minicolumns were perfused online ex vivo in the single pass mode with fresh blood from 8 healthy donors at a flow rate of 6.25 ml/min. The anticoagulation consisted of 0.5 IU heparin plus 0.0-2.1 mg citrate per ml of blood. In Part 1 of the study (40 min perfusion, n = 4), the optimal anticoagulation was found to be 0.5 IU heparin plus about 1 mg citrate per ml of blood. In Part 2 of the study, four 80 min test runs were performed. No signs of hemolysis were found, and the thrombogenicity of the AA gel was negligible. Cell counts and column inlet pressures remained constant; toward the end of the 80 min test-runs, some activation of blood cells (elastase, beta-thromboglobulin), the complement system (C3a, C5a) and the plasmatic coagulation (thrombin-antithrombin complex) was detectable. A moderate initial bradykinin release rapidly subsided to very low levels. In summary, AA minicolumns showed good biocompatibility upon contact with human whole blood and merit further investigation in a closed-loop system for a potential application of direct tumor antibody removal by hemoperfusion. PMID- 11012541 TI - Artificial sensorimotor integration in spinal cord injured subjects through neuromuscular and electrotactile stimulation. AB - Spinal cord injured (SCI) subjects lack sensorimotor functions. Neuromuscular electrical stimulation (NMES) systems have been used to artificially restore motor functions, but without proprioceptive feedback, SCI subjects can control NMES systems only when they can see their limbs. In a gait restoration system, the subject looks down to the ground to be aware of where his foot is while in a grasping activity, maximum grip strength is employed regardless of the force that is required to perform tasks. This report focuses on artificial sensorimotor integration. Multichannel stimulation was used to restore motor functions while encoded tactile sensation (moving fused phantom images) relating to artificially generated movements was provided by electrotactile stimulation during walking and grasping activities. The results showed that the sensorimotor integration attained yielded both the recognition of artificial grasp force patterns and a technique to be used by paraplegics allowing spatial awareness of their limb while walking. PMID- 11012542 TI - Age dependency of neointima formation on vascular prostheses in dogs. AB - Neointima formed quickly on vascular prostheses implanted in young dogs but not in aged dogs. Previously, we found that impeding neointima formation on vascular prostheses occurred more frequently in aged animals. From these observations, we hypothesized that neointima formation was age-dependent in dogs. To test the hypothesis, 26 fabric Dacron vascular prostheses were analyzed. Half of them were retrieved from aged dogs (more than 13 years old) while the other half were from young ones (less than 1 year old). The grafts were harvested at 8 weeks and 3 months after implantation. The graft surfaces were photographed and analyzed by computer for the ratio of the areas with and without thrombus. Light and scanning electron microscopic observation revealed that most of the thrombus-free areas were lined with endothelial cells. Then the endothelialized areas were calculated. Using data obtained from macroscopic, light microscopic, and scanning electron microscopic observations, the arithmetic means were calculated as the degree of neointima formation. In young animals, the degrees at 8 weeks and at 3 months were 89.1 +/- 8.5% (mean +/- SD) and 95.7 +/- 3.3%, respectively. In old animals, they were 27.9 +/- 5.9% and 31.5 +/- 6. 8%, respectively. From these results, we concluded that neointima formation was age-dependent in dogs. PMID- 11012543 TI - Is avoidance of air contact necessary for the in vitro evaluation of thrombogenicity in mechanical circulatory assist devices? AB - An effective in vitro protocol for the investigation of thrombogenicity can provide many advantages in the development of mechanical circulatory assist devices. Strict avoidance of air contact with blood recently was proposed for reliable in vitro evaluation. This study was performed to confirm the necessity of avoidance of air contact for the in vitro test of thrombogenicity in a rotary pump. Two sets of mock circuits with the same rotary blood pumps, reservoirs, and connecting tubes were made. In one system, blood came in contact with air while the other did not. The test blood was heparinized at the dose of 1 IU per 1 ml of blood. The tests were terminated at an activated coagulation time of 1.5 times the control value. The levels of hematocrit, platelet, factors VIII and XII, fibrinogen, thromboxane B2, and plasma-free hemoglobin were measured during the procedures. After the experiments, the thrombi formed were observed, measured, and compared with those formed in in vivo circumstances. The tests were repeated 12 times. There were no statistically significant differences between the 2 groups in hematologic parameters and the amounts of thrombi formed. The thrombi observed in both groups showed the same pathologic findings as those formed in vivo with the exception of intermittent multiple air bubbles found in thrombi of the air-contact group. In conclusion, the effect of air contact in the in vitro investigation of thrombogenicity was negligible while the proposed in vitro test models of thrombogenesis in the mechanical circulatory assist device proved to be reliable. PMID- 11012544 TI - Analytical investigation of leakage flow in disk clearance of a magnetically suspended centrifugal impeller. AB - Leakage flow through the disk clearance of a magnetically suspended centrifugal blood pump is essential for a good washout. An analytical approach, based on the theory of lubrication, is used to predict the leakage volume flow rate, nondimensional radial velocity, nondimensional mean pressure distribution, and comparative shear stress distribution for different disk clearance geometry under varying rotational speeds. The results showed that nondimensional mean pressure distribution and nondimensional radial velocity distribution along the clearance are independent of rotational speed. The flow through the gap depends on a nondimensional parameter S that denotes the ratio of centrifugal forces to the head generating capability of the impeller. It was found that an impeller having a lower S has less possibility of flow reversals in the gap, and gap with maximum height at the outside radius also is more susceptible to flow reversals at the impeller surface. The comparative shear stress within the gap reveals that, in general, the scalar stress is below 500 N/m2. PMID- 11012545 TI - Is erythrocyte damage prevented by gardos effect in hemodialyzed uremic patients? AB - The purpose of this study was to determine the changes of calcium and potassium content in red blood cells (RBC) from uremic patients during a hemodialysis (HD) session. In 17 uremic patients on HD, the calcium and potassium content of RBC was determined in 3 blood samples collected at 0 min-HD (pre-HD), 45 min-HD, and 240 min-HD (end-HD) during a 4 h HD session. The calcium and potassium content of RBC also was determined in 20 normal subjects (controls). The mean values (+/-SD) of RBC calcium content in patients at 0 min-HD, 45 min-HD, and 240 min-HD were 1.95 +/- 0.34, 2.82 +/- 0.50, and 2. 05 +/- 0.4 microg/ml, respectively, and in controls 0.61 +/- 0.14 microg/ml. These values show that the RBC calcium in patients was generally significantly higher (p < 0.0001) in comparison to that of controls. The RBC calcium at 45 min-HD was significantly higher as compared to that at 0 min-HD and at 240 min-HD (p < 0.0001). The mean values (+/-SD) of RBCs' potassium in uremic patients at the previously mentioned measurements were 95.9 +/- 3.34, 92.5 +/- 4.32, and 93.85 +/- 3.89 mEq/L, respectively, and in controls 98.46 +/- 2. 30 mEq/L. These values show that RBC potassium of patients was generally significantly lower in comparison to controls (0 min-HD: p < 0.01, 45 min-HD and 240 min-HD: p < 0.001). Potassium decrease also was significantly lower at 240 min-HD (p < 0.01) and even lower at 45 min-HD (p < 0.001) compared to that at 0 min-HD. In conclusion, uremic patients during an HD session present a high calcium and a low potassium content of erythrocytes. These changes may prevent swelling of the cells (Gardos effect). PMID- 11012546 TI - Serial measurement of polyethylene wear of well-fixed cementless metal-backed acetabular component in total hip arthroplasty: an over 10 year follow-up study. AB - Serial radiographic measurements of polyethylene wear were performed in 38 hips (33 patients) with primary cementless total hip arthroplasty (THA). The average follow-up period was 131.8 months. All prostheses were assessed as radiographically stable at the latest follow-up. A two-dimensional method was used to calculate the relative migration of the femoral head center to the cup center. The average total linear wear and wear rate were 1.22 mm and 0.11 mm/year, respectively. The degree of wear in the first 2 postoperative years accounted for nearly 40% of the total wear at the end of the study (average follow up: 131.8 +/- 10.0 months, +/-SD). The migration of the femoral head at an average period of 3. 4 months after operation accounted for 56% of the amount of wear in the first 2 years. Wear rate decreased gradually with time and stabilized after the fourth year. However, in 2 patients, a progressive increase in the wear rate was associated with severe osteolysis and failure of THA. Both creep and wear contributed to the femoral penetration into the polyethylene liner. The influence of creep cannot be ruled out, especially in the early period after operation. Polyethylene wear is a multifactorial process, and the study of individual wear patterns might be useful in identifying patients who are at risk of late failure of THA. PMID- 11012547 TI - Radiopacity and fatigue characterization of a novel acrylic bone cement with sodium fluoride. AB - Acrylic bone cement must provide good radiographic visibility and good long-term mechanical resistance in joint replacements. A new formulation of cement with 6% barium sulfate and 6% sodium fluoride was developed (Fluoride Bone Cement). Barium sulfate is a necessary addition to allow radiographic visibility although it reduces the mechanical strength of the material. Sodium fluoride promotes bone formation. However, its effect on the mechanical behavior is currently unknown while its influence on radiopacity can only be roughly estimated. The aim of this investigation was to establish if the new formulation would be suitable for clinical trials. In this respect, a mechanical (fatigue test) and radiographic (optical density measurements on x-ray films) characterization was performed on a typical commercially available cement with barium sulfate added and on the Fluoride Bone Cement. It was demonstrated that the fluoride cement has a (marginally) superior fatigue strength and comparable radiopacity to commercial radiopaque cements. PMID- 11012548 TI - Transaortic inflow and outflow cannula for centrifugal mechanical assistance. AB - A specific cannula allowing single site transaortic inflow and outflow cannulation for centrifugal assist devices is described. The cannula is inserted through a straight 18 mm collagen coated Dacron tube anastomosed to the anterior aspect of the ascending aorta. The inflow conduit of the cannula is positioned into the left ventricle through the aortic valve, and the end hole of the outflow conduit is positioned in the ascending aorta. The cannula was evaluated in vivo in 3 adult pigs by the institution of a centrifugal pump for left ventricular support. Optimal flow varied between 5 and 6 L/min and mean aortic pressure between 55 and 70 mm Hg throughout the 3 days of left ventricular support. Recently, we employed this cannula in a patient who was not able to be weaned from cardiopulmonary bypass for the institution of left ventricular support using a centrifugal pump. The cannula provided effective inflow and outflow drainage with an optimal flow of 5.2 L/min throughout the 72 h of support. The patient was successfully weaned from support on the fourth postoperative day. This rational transaortic approach of cannulation using this specific cannula is a refined implantation technique which allows direct left ventricular inflow drainage, reduces the time of implantation, spares left ventricular myocardium, avoids bleeding that is sometimes encountered at other cannulation sites, and avoids compression of the heart by cannulas. PMID- 11012549 TI - Implantation of one piece biventricular assist device by left thoracotomy in an ovine model. AB - In this report, we describe an operative procedure for our implantable 1 piece biventricular assist device (BiVAD) based on a moving actuator mechanism, using an ovine model. Our implantable BiVAD is a volumetric coupled 1 piece unit including right and left blood sacs and an actuator assembly based on the moving actuator mechanism. The BiVAD was controlled by fixed rate control with 75 bpm for the most part. Both the left and the right full ejection modes with the maximum stroke angle were selected to minimize blood stasis in the blood sacs because of low assist flow condition. Three Corriedale sheep were used for the device implantation by a left thoracotomy incision. Cannulation was successfully performed in all cases. Although exposability of the right atrial appendage varied from animal to animal, the insertion of the cannula was easily performed. The cannulas were connected to the pump-actuator assembly in the preperitoneal pocket. All 3 animals survived the experimental procedure. During implantation of the device, in the 1 month survival animal, pump flow was maintained between 2.0 L/min and 2.5 L/min, mean aortic pressure was 90-110 mm Hg, and mean pulmonary artery pressure was 20-30 mm Hg. The left and right atrial pressure were maintained between 0 and 5 mm Hg. In conclusion, this ovine model for implantation of the 1 piece BiVAD can be an effective alternative for testing in vivo biocompatibility of the device although it needs more consideration for anatomical fittability for future human application. PMID- 11012551 TI - Drug-induced proarrhythmic effects: assessment of changes in QT interval. PMID- 11012550 TI - Macrolide - induced clinically relevant drug interactions with cytochrome P-450A (CYP) 3A4: an update focused on clarithromycin, azithromycin and dirithromycin. PMID- 11012553 TI - In vitro evaluation of potential in vivo probes for human flavin-containing monooxygenase (FMO): metabolism of benzydamine and caffeine by FMO and P450 isoforms. AB - AIMS To determine the FMO and P450 isoform selectivity for metabolism of benzydamine and caffeine, two potential in vivo probes for human FMO. METHODS Metabolic incubations were conducted at physiological pH using substrate concentrations of 0.01-10 mM with either recombinant human FMOs, P450s or human liver microsomes serving as the enzyme source. Products of caffeine and benzydamine metabolism were analysed by reversed-phase h.p.l.c. with u.v. and fluorescence detection. RESULTS CYP1A2, but none of the human FMOs, catalysed metabolism of caffeine. In contrast, benzydamine was a substrate for human FMO1, FMO3, FMO4 and FMO5. Apparent Km values for benzydamine N-oxygenation were 60 +/- 8 microM, 80 +/- 8 microM, > 3 mM and > 2 mM, for FMO1, FMO3, FMO4 and FMO5, respectively. The corresponding Vmax values were 46 +/- 2 min-1, 36 +/- 2 min-1, < 75 min-1 and < 1 min-1. Small quantities of benzydamine N-oxide were also formed by CYPs 1A1, 1A2, 2C19, 2D6 and 3A4. CONCLUSIONS: FMO1 and FMO3 catalyse benzydamine N-oxygenation with the highest efficiency. However, it is likely that the metabolic capacity of hepatic FMO3 is a much greater contributor to plasma levels of the N-oxide metabolite in vivo than is extrahepatic FMO1. Therefore, benzydamine, but not caffeine, is a potential in vivo probe for human FMO3. PMID- 11012552 TI - Parkinson's disease and CYP1A2 activity. AB - AIMS MPTP, a neurotoxin which induces parkinsonism is partially metabolized by the enzyme CYP1A2. Smoking appears to protect against Parkinson's disease (PD) and cigarette smoke induces CYP1A2 activity. Thus, we investigated the hypothesis that idiopathic PD is associated with lower CYP1A2 activity using caffeine as a probe compound. METHODS CYP1A2 activity was assessed using saliva paraxanthine (PX) to caffeine (CA) ratios. Caffeine half-life was also estimated from salivary concentrations of caffeine at 2 and 5 h post dose. 117 treated and 40 untreated patients with PD and 105 healthy control subjects were studied. RESULTS PX/CA ratios were 0. 57, 0.93 and 0.77 in treated patients, untreated patients and healthy control subjects, respectively, with no significant differences between study groups (95% CI: treated patients vs controls -0.24, 0.57; untreated patients vs controls -0.75, 0.35). However, patients with PD (treated or untreated) had caffeine half-lives shorter than that in controls (treated patients: 262 min, untreated patients: 244 min, controls: 345 min; 95% CI: controls vs treated patients 23, 143 (P = 0.003); controls vs untreated patients 19, 184 (P = 0.011)). Amongst the patients with PD, caffeine half-life was also inversely related to the age of onset of disease (P = 0.012); gender and concomitant drugs did not influence this significantly. CONCLUSIONS: Based on PX/CA ratio, there was no evidence of decreased CYP1A2 activity in patients compared with control subjects. The observed decrease in the elimination half life of caffeine in PD may be caused by increased CYP2E1 activity, an enzyme that also contributes to the metabolism of caffeine. The latter warrants further investigation. PMID- 11012554 TI - Population pharmacokinetics of doxorubicin, etoposide and ifosfamide in small cell lung cancer patients: results of a multicentre study. AB - AIMS: To determine the population pharmacokinetic (PK) parameters of doxorubicin (Dox), etoposide (Eto) and ifosfamide (Ifo) in small cell lung cancer (SCLC) patients, to assess the potential relationship between those parameters and to estimate the impact of individual morphological and biological covariates on patients' PK parameters. METHODS: Twenty-four patients with either SCLC limited to the thorax or extensive SCLC entered the study. All but one received at least two 3 day courses of the standard AVI (Dox 50 mg m-2 day 1, Eto 120 mg m-2 day 1,2,3, Ifo 2000 mg m-2 day 1,2) regimen. Individual blood samples were collected during each course and data on 47 courses were available. Data were analysed with the NONMEM program. Dox, Eto and Ifo plasma concentrations were studied with multicompartment (3, 2 and 2, respectively) models. Inter-individual and interoccasion (course-to-course) variabilities were estimated. The influence of individual covariates (age, sex, stage of the disease, weight, height, body surface area, serum creatinine, total protein, LDH, ASAT, ALAT, alkaline phosphatase, gamma-GT, bilirubin) on PK parameters was also assessed. Correlations between individual PK parameters of Dox, Eto and Ifo were explored by using Pearson's correlation coefficient. RESULTS: Multiple data were available for each patient. Dox clearance (CL) and volume of distribution (Vd) were 32.0 l h-1 and 9.3 l (Inter-individual variability: 17.2% and 19.2%). Eto CL (l h-1) and Vd were, respectively, 3.34-0.0083* serum creatinine (micromol l-1) and 6.38 l (interindividual variability: 15.6% and 18.7%). Ifo CL and Vd at day 1 were 5.6 l h-1 and 26.0 l (interindividual variability: 10.1% and 17.2%, respectively). Estimation of course-to-course variability improved the precision of PK models in some cases. No correlation was observed between the respective PK parameters of each drug. Of individual covariates tested, only serum creatinine correlated with Eto CL (r = -0.37, P < 0.001). Self-induction of the metabolism of Ifo was apparent (mean CL increase from day 1 to day 2 : 42%) and individually correlated with the CL value at day 1 (r = -0.61, P < 0.001). CONCLUSIONS: Assessment of potential relationships between individual systemic exposure of chemotherapy and therapeutic endpoints (tumour response, toxicity and survival) will be required to adjust drugs dosages based on individual PK parameters rather than questionable body-surface area. However, all three drugs in the AVI regimen should be monitored simultaneously. PMID- 11012555 TI - Effect of altered gastric emptying and gastrointestinal motility on metformin absorption. AB - AIMS: The purpose of this in vivo human study was to assess the effect of altered gastric emptying and gastrointestinal motility on the absorption of metformin in healthy subjects. METHODS: An open-label, three treatment, three period crossover study was conducted in 11 healthy volunteers. Each subject received 550 mg metformin hydrochloride in solution alone; 5 min after a 10 mg i.v. dose of metoclopramide; and 30 min after a 30 mg oral dose of propantheline. Metformin solution was radiolabeled by the addition of 99mTc-DTPA. The gastrointestinal transit of the solution was monitored by gamma scintigraphy and the pharmacokinetic data were correlated with the scintigraphic findings. RESULTS: Scintigraphic data indicated that pretreatment with metoclopramide decreased gastric emptying time and increased gastrointestinal motility while pretreatment with propantheline had the opposite effect. The systemic disposition of metformin was not altered by pretreatment with metoclopramide and propantheline, as judged by unchanged renal clearance and elimination half-life of metformin. Extent of metformin absorption was essentially unchanged after pretreatment with metoclopramide. However, AUC(0,infinity) and % UR (percent dose excreted unchanged in urine) generally increased with increase in gastric emptying time and small intestinal transit times. GI overlay plots showed that the absorption phase of metformin plasma profile always coincided with gastric emptying and the beginning of decline of metformin plasma concentrations was usually associated with the colon arrival. Only in cases where the intestinal transit was drastically prolonged by propantheline pretreatment, was a decline in plasma levels observed prior to colon arrival. CONCLUSIONS: Metformin is primarily absorbed from the small intestine. The extent of metformin absorption is improved when the gastrointestinal motility is slowed. These findings have significant implications in the design of a metformin modified release dosage form. PMID- 11012556 TI - Effect of an oral contraceptive preparation containing ethinylestradiol and gestodene on CYP3A4 activity as measured by midazolam 1'-hydroxylation. AB - AIMS: To characterize the effect of an oral contraceptive (OC) containing ethinylestradiol and gestodene on the activity of CYP3A4 in vivo as measured by the 1'-hydroxylation of midazolam. METHODS: In this randomised, double-blind, cross-over trial nine healthy female subjects received either a combined OC (30 microg ethinylestradiol and 75 microg gestodene) or placebo once daily for 10 days. On day 10, a single 7.5 mg dose of midazolam was given orally. Plasma concentrations of midazolam and 1'-hydroxymidazolam were determined up to 24 h and the effects of midazolam were measured with three psychomotor tests up to 8 h. RESULTS: The combined OC increased the mean AUC of midazolam by 21% (95% CI 2% to 40%; P = 0.03) and decreased that of 1'-hydroxymidazolam by 25% (95% CI 10% to 41%; P = 0.01), compared with placebo. The metabolic ratio (AUC of 1' hydroxymidazolam/AUC of midazolam) was 36% smaller (95% CI 19% to 53%; P = 0.01) in the OC phase than in the placebo phase. There were no significant differences in the Cmax, tmax, t(1/2) or effects of midazolam between the phases. CONCLUSIONS: A combined OC preparation caused a modest reduction in the activity of CYP3A4, as measured by the 1'-hydroxylation of midazolam, and slightly increased the AUC of oral midazolam. This study suggests that, at the doses used, ethinylestradiol and gestodene have a relatively small effect on CYP3A4 activity in vivo. PMID- 11012557 TI - A double-blind, placebo-controlled study to assess tolerability, pharmacokinetics and preliminary pharmacodynamics of single escalating doses of Z13752A, a novel dual inhibitor of the metalloproteases ACE and NEP, in healthy volunteers. AB - AIMS: The objective of this study was to evaluate the tolerability of a novel dual ACE-NEP inhibitor, Z13752A, after the oral administration of rising single doses in healthy volunteers. This study was also a preliminarily investigation of Z13752A pharmacodynamics (PD) and pharmacokinetics (PK). METHODS: In this randomized, placebo-controlled, sequential study, two alternating panels of eight healthy male volunteers each (six subjects receiving the active treatment + two subjects receiving placebo) were treated with increasing oral doses of Z13752A: 10, 50, 200, and 600 mg were given to panel I and 20, 100, 400 and 800 mg were given to panel II. The study was double-blind relative to placebo or active treatment, and was open with respect to the dose levels. The same volunteer received placebo only once. RESULTS: Single oral doses of Z13752A, as high as 800 mg, were well tolerated. Only six mild-to-moderate adverse events mainly headache, were reported and appeared to be of little clinical relevance. After administration of 200, 400, 600 and 800 mg of Z13752A, a nonsignificant fall in diastolic blood pressure was detected, in both the standing and supine position. After single oral doses of Z13752A, ACE inhibition appeared to be significant at all the doses tested, linearly correlated with the dose and was almost complete at doses > or = 100-200 mg. NEP inhibition was indicated by elevation of ANP and cGMP plasma concentrations in almost all subjects. In the 200-800 mg dose range, Z13752A produced a 50-100% increase of plasma cGMP levels and a 50-80% elevation in urinary cGMP concentrations. Detectable plasma levels of Z13752A were found in all the treated subjects. Z13752A was well and rapidly absorbed, with peak concentrations reached approximately 2.5 h after administration. The mean apparent elimination half-life from plasma was approximately 12 h. The pharmacokinetics of Z13752A after single oral doses were characterized by low intersubject variability and appeared to be dose-independent. CONCLUSIONS: Z13752A showed a good single dose tolerability profile at doses up to 800 mg. The pharmacokinetic data indicate that Z13752A administered orally is rapidly absorbed and available to the systemic circulation in humans. The relatively slow clearance indicates that a once-a-day dose regimen could be considered for Z13752A. PMID- 11012558 TI - Pharmacodynamic and pharmacokinetic profile of S 17092, a new orally active prolyl endopeptidase inhibitor, in elderly healthy volunteers. A phase I study. AB - AIMS: The aim of this study was to characterize the pharmacodynamics and the pharmacokinetics of S 17092, a new orally active prolyl endopeptidase inhibitor following single and repeated administration in elderly healthy volunteers. METHODS: This was a double-blind, randomized, placebo-controlled, single and multiple dose study in elderly healthy male and female volunteers (n = 36). Four doses were investigated in sequential order: 100, 400, 800 and 1200 mg. Each dose was administered orally once a day in single administration and then, after a 1 week washout period, during 7 days. Pharmacodynamics were assessed by measurement of plasmatic prolyl endopeptidase (PEP) activity, quantitative electroencephalogram (EEG) and psychometric tests. S 17092 concentrations in plasma were quantified by high performance liquid chromatography with tandem mass spectrometric detection. RESULTS: PEP activity in plasma was dose-dependently inhibited both after administration of a single dose and after repeated doses of S 17092. The mean maximal inhibition was obtained within 0.5-2 h after dosing, while inhibition lasted at least 12 h after dose administration. S 17092 appeared to be a centrally active substance as it induced statistically significant modifications in EEG compared with placebo. S 17092 at 100 mg exerted an acute increase in alpha band following single administration at 4 h and 8 h postdosing. When administered repeatedly over 7 days S 17092 did not appear to induce significant lasting central nervous system (CNS) effects. In psychometric tests, response times in the numeric working memory were significantly reduced compared with placebo, following the 800 mg dose. There were some beneficial residual effects of the 1200 mg dose on day 13: delayed word recall and word recognition sensitivity improved compared with the declines noted under placebo. Maximum measured concentration (Cmax) and area under the curve (AUC) parameters increased in proportion to the dose. The terminal half-life (t(1/2)) values ranged between 9 and 31 h on day 1 and between 7 and 18 h on day 14. A high interindividual variability was observed at all dose levels. S 17092 was well tolerated with no clinically significant changes in laboratory or physical parameters observed at any dose. CONCLUSIONS: S 17092 had a potent, dose-dependent inhibitory effect on plasmatic PEP, increased alpha band EEG at the 100 mg dose and improved performance in two verbal memory tests at the 1200 mg dose while there were disruption to the vigilance task. The results obtained in elderly healthy subjects indicated that S 17092 is suitable for once-daily dosing without any serious adverse events. PMID- 11012559 TI - Effects of high ambient temperature on parasympathetically mediated cardiovascular reflexes in normal man. AB - AIMS: To examine the effects of high ambient temperature ('heat stressor') on parasympathetically mediated cardiovascular reflexes (power of respiratory sinus dysrhythmia; change in heart rate elicited by change in posture from lying to standing ['30 : 15 ratio']). METHODS: Twelve healthy male volunteers participated in four weekly sessions, each of which was associated with one treatment condition (placebo at an ambient temperature of 22 degrees C; propranolol 40 mg at 22 degrees C; placebo at 40 degrees C; propranolol 40 mg at 40 degrees C), according to a balanced double-blind design. Heart rate was recorded by ECG, finger tremor (7-12 Hz) with an accelerometer strapped to the middle finger of the nondominant hand, and sublingual temperature by a mercury thermometer. Power of finger tremor and the variations of the R-R intervals of the ECG were obtained from Fourier transformations of the data. Data were analysed by analysis of variance, with repeated measures using a significance criterion of P < 0.05; individual comparisons of active treatment with placebo and of data obtained at 40 degrees C with those obtained at 22 degrees C were made with Fisher's Least Significant Difference test. RESULTS: Heart rate was increased by the heat stressor, and this increase was abolished by propranolol. The heat stressor reduced the power of respiratory sinus dysrhythmia and the 30 : 15 ratio, and increased the power of physiological finger tremor. Propranolol did not affect heat stressor-induced changes in the parasympathetic cardiac reflexes, but reduced the heat stressor-induced enhancement of finger tremor. CONCLUSIONS: The increase in the power of physiological finger tremor at high ambient temperature is consistent with sympathetic activation, whereas the reduction in the power of respiratory sinus dysrhythmia and 30 : 15 ratio indicates a decrease in parasympathetic activity. These results demonstrate that high ambient temperature may induce vagal withdrawal in the heart. PMID- 11012560 TI - The rates of common adverse events reported during treatment with proton pump inhibitors used in general practice in England: cohort studies. AB - AIMS: To estimate the rates of common adverse events in patients treated with the proton pump inhibitors omeprazole, lansoprazole and pantoprazole in general practice in England. METHODS: In prescription-event monitoring cohort studies, data on dispensed prescriptions prescribed by general practitioners in England soon after each drug was launched were linked to subsequent clinical events recorded by the prescriber. 16 205 patients prescribed omeprazole between June 1989 and June 1990, 17 329 patients prescribed lansoprazole between May and November 1994, and 11 541 patients prescribed pantoprazole between December 1996 and June 1997 were studied. RESULTS: The commonest adverse events in the omeprazole, lansoprazole and pantoprazole cohorts were diarrhoea (incidence: 0. 18, 0.39 and 0.23 per 1000 days of exposure, respectively); nausea/vomiting (incidence: 0.16, 0.22 and 0.18 per 1000 days of exposure, respectively); abdominal pain (incidence: 0.17, 0.21 and 0. 17 per 1000 days of exposure, respectively); and headache (incidence rates: 0.10, 0.17 and 0.15 per 1000 days of exposure, respectively). The remaining adverse events occurred at rates of less than 0.11 per 1000 days of exposure. There were little absolute differences in the rates of most events between the three proton pump inhibitors. However, diarrhoea was more commonly associated with lansoprazole compared with omeprazole (rate difference: 0.21 per 1000 days of exposure; 95% CI 0.17, 0.25; rate ratio: 2.11; 1.78, 2.51), and there was a clear age-response relationship. CONCLUSIONS: Adverse events occurred relatively infrequently in all three cohorts. There were only small absolute differences in event rates between the three drugs, although these data suggest the hypothesis that lansoprazole is associated with more frequent occurrence of diarrhoea, particularly in the elderly. PMID- 11012561 TI - The pharmacokinetics of trinitroglycerin and its metabolites in patients with chronic stable angina. AB - AIMS: To study the pharmacokinetics of trinitroglycerin (GTN) and its four metabolites in angina patients during their transient use of transdermal GTN tape following intravenous administration of GTN. METHODS: Four patients received a GTN tape following intravenous administration of 0.1 microg kg-1 min-1 GTN, and the other four patients received two GTN tapes following intravenous administration of 0.2 microg kg-1 min-1 GTN. RESULTS: Plasma concentrations of GTN in both groups during tape application showed a slight decrease for the hour after the application of the tape and then were con- stant for 24 h. In contrast, the concentrations of dinitroglycerins (GDNs) and mononitroglycerins (GMNs) depended on the duration of previous intravenous administration of GTN. Neither significant cardiovascular changes nor undesirable complications were observed during the study. CONCLUSIONS: The results suggest that appropriate replacement of intravenous GTN administration with transdermal tape application could maintain a therapeutic GTN level. PMID- 11012562 TI - Nebivolol decreases systemic oxidative stress in healthy volunteers. AB - AIMS: Nebivolol is a selective, vasodilatory beta1-adrenergic receptor antagonist which has been suggested to possess additional antioxidative properties. The aim of the present study was to assess the actions of nebivolol in antihypertensive doses on systemic oxidative stress in healthy volunteers, reflected by 24 h urinary excretion of 8-iso-PGF2alpha. METHODS: In a double-blind, cross-over study, 12 healthy volunteers received 5 mg nebivolol once daily or placebo for a total of 7 days, separated by a wash out period of 2 weeks. After each treatment period 24 h urinary excretion of 8-iso-PGF2alpha was determined by gas chromatography-tandem mass spectrometry. RESULTS: After the 7 day treatment period nebivolol decreased significantly urinary excretion of 8-iso-PGF2alpha by 24% from 55.3 +/- 5.1 pmol mmol-1 creatinine during the placebo period to 42.3 +/ 4.7 pmol mmol-1 creatinine (mean +/- s.e. mean, P = 0. 01), a mean decrease of 13 pmol mmol-1 creatinine (95% CI: -22.8; -3. 1). CONCLUSIONS: Our data show for the first time that nebivolol decreases systemic oxidative stress in young healthy volunteers. PMID- 11012563 TI - Influences on plasma cortisol of different formulations of beclomethasone dipropionate. PMID- 11012564 TI - Recent advances in the molecular genetics of congenital and acquired primary adrenocortical failure. PMID- 11012565 TI - Characterization of a novel DNA polymorphism in the human CYP21 gene and application for DNA diagnosis of congenital adrenal hyperplasia. AB - OBJECTIVES: Congenital adrenal hyperplasia (CAH) is a common endocrine disorder and CYP21 (21-OH, EC 1.14.99.10) deficiency is the most common cause of the disease. The presence of a pseudogene and a wide range of mutation types often makes DNA diagnosis difficult. Analysis of mutant alleles from patients with CAH identified a new DNA polymorphism in exon 10. To test the usefulness of this polymorphism, linkage analysis was performed using three RFLP's at both end of the CYP21 gene in patients and controls. DESIGN AND PATIENTS: Genomic DNA was extracted from 21 unrelated patients and 39 unaffected individuals. Haplotyping analysis was performed for three RFLPs, MspI and Fnu4HI in intron 2 and a new SmaI RFLP in exon 10. All three polymorphic sites were characterized by DNA sequencing and usefulness of these RFLPs for DNA diagnosis was tested in patients' families. MEASUREMENT: CAH patients were diagnosed by clinical symptoms and biochemical tests. Allele frequencies and heterozygosities were studied for three RFLP's in patients and controls using polymerase chain reaction (PCR). RESULTS: A new SmaI RFLP showed a sequence difference as G or A at the nucleotide position 2694 in exon 10. Sequences at the MspI and Fnu4HI polymorphic sites were T or C at the nucleotide position 395 and 453, respectively. All of these RFLPs showed biallelic DNA polymorphisms and codominant segregation in family analysis. Heterozygosities were 0.31 for MspI, 0.48 for Fnu4HI and 0.44 for SmaI in normal individuals. There was no linkage disequilibrium for three RFLPs between patients and controls. CONCLUSIONS: The SmaI RFLP can be useful for linkage analysis and DNA diagnosis in conjunction with RFLPs in intron 2 in CAH families because the polymorphic site is within the active gene at the 3' end. DNA sequencing results revealed that these RFLPs were created by gene conversions as with other mutations in the CYP21 gene. PMID- 11012566 TI - Tall stature in familial glucocorticoid deficiency. AB - OBJECTIVE: Familial glucocorticoid deficiency (FGD) has frequently been associated with tall stature in affected individuals. The clinical, biochemical and genetic features of five such patients were studied with the aim of clarifying the underlying mechanisms of excessive growth in these patients. PATIENTS AND METHODS: Five patients with a clinical diagnosis of FGD are described in whom the disorder resulted from a variety of novel or previously described missense or nonsense mutations of the ACTH receptor (MC2-R). All patients demonstrated excessive linear growth over that predicted from parental indices and increased head circumference. RESULTS: Growth hormone and IGF-I values were normal. Growth charts suggest that the excessive growth is reduced to normal following the introduction of glucocorticoid replacement. A characteristic facial appearance including hypertelorism, marked epicanthic folds and prominent frontal bossing was noted. CONCLUSIONS: These findings indicate that ACTH resistance resulting from a defective ACTH receptor may be associated with abnormalities of cartilage and/or bone growth independently of the GH-IGF-I axis, but probably dependent on ACTH actions through other melanocortin receptors. PMID- 11012567 TI - Comparison of one week 0900 h serum cortisol, low and standard dose synacthen tests with a 4 to 6 week insulin hypoglycaemia test after pituitary surgery in assessing HPA axis. AB - OBJECTIVE: To compare the use of 0900 h serum cortisol and both low and standard dose Synacthen tests, one week after pituitary surgery with an insulin hypoglycaemia test performed 4-6 weeks after surgery in assessing the integrity of the hypothalamic-pituitary-adrenal (HPA) axis. DESIGN: 0900 h basal serum cortisol was measured on days 6 and 7 after pituitary surgery (24 h off replacement hydrocortisone) followed by a low dose Synacthen test (1 microg intravenously) on day 6 and a standard dose Synacthen test (250 microg intramuscularly) on day 7. Three to 5 weeks later an insulin hypoglycaemia test was performed on all patients. Both low and standard dose Synacthen tests were performed on control subjects using an identical protocol. SUBJECTS: Forty-two patients (21 male, 21 female), median age 49 years (range 23-73) who had pituitary surgery (Cushing's disease excluded). One patient had undergone repeat surgery for residual tumour and was studied following each operation. Sixteen healthy normal volunteers, median age 37 years (range 21-55). MEASUREMENTS: Serum cortisol measured by radioimmunoassay. RESULTS: Two standard deviations below the mean serum cortisol (logarithmic transformation) in the normal volunteers 30 minutes after low dose Synacthen (1 microg) was 496 nmol/l and after standard dose Synacthen (250 microg) was 504 nmol/l. A normal response was therefore taken as serum cortisol > 500 nmol/l at 30 minutes in both tests (using 496 and 504 nmol/l did not alter results). 0900 h serum cortisols 1 week after surgery were > 250 nmol/l in 31 patients and 29 of these had a normal response to hypoglycaemia (peak serum cortisol > 550 nmol/l). Of the remaining two patients, one had 0900 h serum cortisol on day 6 and 7 after surgery of 405 and 441 nmol/l with a peak serum cortisol response to hypoglycaemia of 451 nmol/l; the other patient had 0900 h serum cortisols of 416 and 251 nmol/l and a peak cortisol response to hypoglycaemia of 498 nmol/l. All eight patients who had a 0900 h serum cortisol < 100 nmol/l failed a subsequent insulin hypoglycaemia test. Seven discrepancies were noted between the low dose Synacthen test and the insulin hypoglycaemia test in the 41 patients who had both tests. In six of these, a subnormal response to low dose Synacthen was followed by a normal response to hypoglycaemia. Three discrepancies were noted between the standard dose Synacthen test and the insulin hypoglycaemia test in the 40 patients who had both tests. In all three cases a normal response to Synacthen was followed by a subnormal response to hypoglycaemia. CONCLUSIONS: A 0900 h serum cortisol < 100 nmol/l (24 h off replacement hydrocortisone) indicated ACTH deficiency and need for lifelong steroid replacement. A 0900 h serum cortisol > 450 nmol/l one week after pituitary surgery is highly suggestive of a normal cortisol response to hypoglycaemia. A 0900 h serum cortisol between 250 and 450 nmol/l one week after pituitary surgery permits safe withdrawal of steroid therapy pending an insulin hypoglycaemia test 1 month after surgery. Patients with 0900 h serum cortisol between 100 and 250 nmol/l should continue replacement steroids until definitive testing. Low dose and standard dose Synacthen tests 1 week after pituitary surgery are unreliable and should not be used. PMID- 11012568 TI - Adrenal responsiveness to high, low and very low ACTH 1-24 doses in obesity. AB - OBJECTIVE: To investigate adrenal activity in visceral obesity in which adrenal hyperactivity has been hypothesized. This could reflect hypothalamus-pituitary alterations leading to slight hyperfunction of the adrenal. Primary adrenal hypersensitivity to ACTH drive in obesity has also been suggested. However, it has also been reported that dehydroepiondrosterone (DHEA) levels in obesity are reduced and it has been hypothesized that this could play a role in the increased cardiovascular risk in obese patients. SUBJECTS: We have studied seven obese women with visceral adiposity (OB, age: 33.6+/-3.3 years, BMI: 33.8+/-1.3 kg/m2, WHR: 0.88+/-0.01). The results in OB were compared with those recorded in a group of age-matched normal women (NS, age: 30+/-1.3 years, BMI: 19.9+/-0.4 kg/m2, WHR: 0.76+/-0.02). METHODS: We have studied the cortisol (F), aldosterone (A) and DHEA responses to ACTH 1-24 administered at low (LD, 0.5 microg/m2) or very low (VLD, 0.125 microg/m2) dose followed by a second challenge with supramaximal dose (HD, 250 microg). RESULTS: Basal F, A and DHEA levels in OB were similar to those in NS. The peak F responses to ACTH were dose-related in both groups. At each dose the F peaks in OB (VLD: 495.6+/-43.9 nmol/l, HD: 722.3+/-67.7 nmol/l; LD: 519.2+/ 46.0 nmol/l, HD: 729.6+/-44.7 nmol/l) were similar to those in NS (VLD: 556.7+/ 45.9 nmol/l, HD: 704.8+/-20.7 nmol/l; LD: 511.8+/-22.8 nmol/l, HD: 726.7+/-26.5 nmol/l). The peak A responses to ACTH were dose-related in both groups. At each dose, the A peaks in OB (VLD: 0.55+/-0.03 pmol/l, HD: 0.79+/-0.09 pmol/l; LD: 0.63+/-0.04 pmol/l, HD: 0.78+/-0.09 pmol/l) were similar to those in NS (VLD: 0.8+/-0.10 pmol/l, HD: 0.86+/-0.09 pmol/l; LD: 0.8+/-0.10 pmol/l, HD: 0.95+/-0.12 pmol/l). The peak DHEA responses to ACTH were dose-related in both groups. At each dose the DHEA peaks in OB (VLD: 58.6+/-13.3 nmol/l, HD: 61.9+/-13.1 nmol/l; LD: 55.18+/-6.4 nmol/l, HD: 72.3+/-9.8 nmol/l) were similar to those in NS (VLD: 54.3+/-8.2 nmol/l, HD: 57.8+/-8.2 nmol/l; LD: 42.2+/-3.7 nmol/l, HD: 56.9+/-4.3 nmol/l). CONCLUSIONS: This study shows that the cortisol, aldosterone and dehydroepiondrosterone responses to high, low and very low ACTH doses in obese women overlap with those in age-matched lean controls; these findings suggest normal sensitivity of the different zones of the adrenal cortex to ACTH in obesity. PMID- 11012569 TI - Vascular reactivity in acromegalic patients: preliminary evidence for regional endothelial dysfunction and increased sympathetic vasoconstriction. AB - BACKGROUND AND OBJECTIVES: Hypertension is found in one-third of acromegalic patients. An heterogenous distribution of cardiac output has been recently demonstrated in acromegalic patients with an increased blood flow at the level of the upper limb, suggesting that acromegalic patients may have some degree of endothelial dysfunction. Elsewhere, studies involving hypopituitary GH-deficient adults have shown that GH and/or IGF-I may have direct effect on endothelial function. SUBJECTS AND METHODS: We thus compared cutaneous vasoreactivity responses in 10 normotensive patients with active acromegaly (A) (six women and four men) aged 25-59 (mean, 43.2 years), whose basal GH and IGF-I levels ranged from 7.4 to 158 mU/l and from 401 to 1690 microg/l, respectively, and in 10 normal age- and sex-matched controls (NC) by means of Laser Doppler flowmetry at the levels of the palm and the dorsum of the right hand. Circulatory skin velocities were studied basally and after increasing skin temperature to 44 degrees C (in order to study direct nonspecific vasodilatation response which is independent of endothelial or autonomous nervous system and reflects normal vascular muscle function), after shear-stress (known to produce flow-dependent vasodilatation, mediated by nitric oxyde (NO) originating from endothelial cells) and after cold-stress applied on the opposite hand (known to produce vaso constriction mediated by the sympathetic nervous system). RESULTS: The warm test induced a significant (P<0.001) and similar increase in both dorsal and palmar skin perfusion in A (mean +/- SD) (240+/-96 and 238+/-134%, respectively) and NC (232+/-137 and 233+/-73, respectively). Ischaemia release induced a significant increase in both dorsal and palmar skin blood flows in the two groups (P<0.001), but reactivities in acromegalic patients were about one half of those measured in controls (22.9+/-16.2% (A) vs. 46.9 25% (NC), 2P<0.02, at the level of the dorsum; and 45.0+/-43.6% (A) vs. 104.7+/-40.1 (NC), 2P<0.01, at the level of the palm). Cold pressor test resulted in significant decreases in both cutaneous flows (P<0.01) in the two groups, with a larger vasoconstriction (that did not reach statistical significance) in acromegalic patients as compared with controls (P< 0.10). CONCLUSION: Vascular smooth cell ability to produce skin vasodilatation is normal but endothelium-dependent vasodilatation appears to be impaired while sympathetic-mediated vasoconstrictive response might be increased in acromegaly. This endothelial dysfunction may contribute to hypertension and represent a risk factor for cardiovascular complications in acromegaly. PMID- 11012570 TI - Sustained reduction in circulating cholesterol in adult hypopituitary patients given low dose titrated growth hormone replacement therapy: a two year study. AB - OBJECTIVE: To study the effects of short (6 months) and longer-term (up to 24 months) growth hormone (GH) replacement therapy using a dose titration regimen, on lipid and glucose metabolism in GH-deficient, hypopituitary adults. DESIGN: On going open study of GH treatment up to 24 months. Measurements were performed at baseline and at 6, 12, 18 months and 2 years during therapy (data shown at 6 months and 2 years only). Using a dose titration regimen the median GH dose used to achieve and maintain IGF-I levels above the median, but below the upper limit of the age-related reference range (median IGF-I 202.5 microg/l, range 76-397 microg/l), was 1.2 IU daily (range 0.4-3 IU) [0.8 IU/day, males; 1.6 IU/day, females]. PATIENTS: Ninety GH-deficient hypopituitary adults (54 female, median age 48 years, range 19-79 years) entered the study and 24 (14 female, median age 45 years, range 32-79 years) have concluded the 2 year period of assessment. MEASUREMENTS: Body mass index (BMI), waist and hip circumference ratio (WHR), fasting lipids, glucose and glycated haemoglobin (HbA1c) levels were measured at 6 month intervals during GH therapy. RESULTS: Using the dose titration regimen, compared to pretreatment values, total and low density lipoprotein (LDL) cholesterol levels were significantly lower at 6 months (mean +/- SEM, 5.61+/-0.1 vs. 5.25+/-0.1, and 3.85+/-0.19 vs. 3.43+/-0.26, respectively, P<0.05), and were maintained throughout the study. Male patients had significantly lower pretreatment total and LDL cholesterol levels than females (mean +/- SEM, 5.33+/ 0.16 mmol/l vs. 5.7+/-0.12 mmol/l and 3.8+/-0.23 mmol/l vs. 3.92+/-0.29 mmol/l, respectively, P< 0.05). A decrease in total cholesterol was confined to patients with pretreatment total cholesterol levels above 5.8 mmol/l; patients with the highest pretreatment cholesterol levels (> 6.4 mmol/l) obtained the greatest cholesterol reduction (mean +/- SEM, 7.13 +/- 0.14 mmol/l vs. 5.76+/-0.31 mmol/l, P<0.05). A cholesterol-lowering effect of GH therapy was evident in patients who had elevated pre-GH total cholesterol levels even if they were already receiving and continuing lipid lowering medication (mean +/- SEM, 5.62+/-0.22 vs. 5.03+/ 0.285, P<0.05). A modest increment in high density lipoprotein (HDL)-cholesterol was evident at 18 months but there was no significant change in triglycerides at any time point. Fasting plasma glucose increased significantly at 6 months but remained within the reference range. Glycated haemoglobin increased significantly at 6 months and was maintained throughout the study; one patient developed frank diabetes mellitus while receiving treatment. There was a weak but significant correlation between the increment in glycated haemoglobin and pretreatment BMI (r = + 0.215, P<0.05). CONCLUSION: The effect of GH on lowering total and low density lipoprotein-cholesterol is more prominent in patients with higher pretreatment cholesterol levels and is evident even in patients receiving other lipid-lowering medication. A modest increment in mean fasting glucose (within the reference range) and mean glycated haemoglobin persisted throughout the study. One patient developed diabetes mellitus. A GH replacement regimen using low dose and careful titration to avoid elevated IGF-I levels and adverse effects is associated with sustained beneficial effects on circulating lipids. PMID- 11012572 TI - Expression of thyroid receptor isoforms in the human fetal central nervous system and the effects of intrauterine growth restriction. AB - BACKGROUND: Congenital hypothyroidism is known to be associated with mental retardation which, if recognized promptly, is largely prevented by thyroid hormone replacement. Intrauterine growth restriction (IUGR) is a major cause of perinatal mortality and morbidity, and is also associated with neurodevelopmental delay. Fetuses with IUGR have reduced circulating concentrations of free thyroxine (T4) and free triiodothyronine (T3), leading to the hypothesis that a reduction in the tissue effects of thyroid hormones in the central nervous system (CNS) may contribute to neurodevelopmental morbidity. Since thyroid hormone effects are mediated through binding to specific nuclear thyroid hormone receptors (TRs), we have defined the pattern of TR isoform expression in the CNS throughout normal human development and have compared TR expression in the CNS of normal fetuses with those affected by IUGR. METHODS: Samples of normal human fetal brain from first and second trimesters were obtained at surgical termination of pregnancy. Appropriately grown and third trimester fetuses affected by Intrauterine growth restriction (IUGR) were also investigated after unexplained stillbirth at post mortem examination. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to examine the expression of TR isoform mRNAs in frozen cerebral cortex from 10 to 16 weeks gestation. TR protein expression in human fetal brains (both cerebral hemispheres and cerebellum) was also examined in formalin fixed sections and expression of TR alpha1, alpha2, beta1 and beta2 isoforms being defined using semiquantitative immunocytochemistry. RESULTS: RT-PCR revealed the presence of mRNAs encoding TR alpha1, beta1 and beta2 isoforms and the nonfunctional TRalpha2 variant in the fetal cerebral cortex from week 10 of human pregnancy. Immunostaining of the fetal brain revealed TR alpha1 and alpha2 protein from week 11 of human gestation. Expression of all TR isoform proteins was largely confined to the pyramidal neurones of the cerebral cortex and the Purkinje cells of the cerebellum with increasing receptor expression evident with gestational age. Semiquantitative observer scoring showed that by the second trimester, there was a marked increase in the proportion of pyramidal and Purkinje cells expressing TR isoforms, while by the third trimester, all these cells immunostained. Comparison of TR immunostaining in the cerebral cortex and cerebellum from IUGR fetuses (n = 18) matched for gestational age to normal fetuses revealed a lower intensity of expression of all the TR isoforms confirmed by observer scoring and quantification using TR protein immunofluoresence (P<0.01). CONCLUSIONS: Our findings indicate both pre- and post-translational expression of TR alpha and beta isoforms in the cerebral cortex of first trimester fetuses. These findings support the view that the transplacental passage of thyroid hormone in early gestation may be critical to neurological development. Our finding that in severe IUGR the expression of TR isoforms in the human fetal cerebral cortex and cerebellum was significantly reduced, in association with reduced circulating thyroid hormone concentrations indicate that changes in free ligand and receptors may affect CNS development. These findings should prompt further investigation of the potential therapeutic role of peripartum thyroid hormone treatment. PMID- 11012571 TI - Functional characterization of five constitutively activating thyrotrophin receptor mutations. AB - OBJECTIVE: Gain of function mutations of the thyrotrophin receptor (TSHR) affect several functional characteristics, such as cAMP and inositol phosphate (IP) accumulation, cell surface expression and TSH affinity. In this study we compared five constitutively activating TSHR mutations, four receptors with a point mutation (S505N, L629F, I630L, V656F) and a nine amino acid (aa) deletion mutant (aa positions 613-621) for these functional parameters in parallel transfection experiments. METHODS: The wild-type TSHR (wt) and TSHRs containing the mutations S505N, L629F, I630L, V656F and the deletion 613-621 (all cloned in the expression vector pSVL) were transiently expressed in COS-7 cells in parallel experiments. Forty-eight hours after transfection the basal and stimulated cAMP and inositol phosphate accumulation as well as the cell surface expression (by FACS and ELISA), KD-values and TSHR down regulation by different stimuli were determined. RESULTS: In contrast to the very different values for specific constitutive activity (sca) (ranging from 7.5 to 100.3-fold wt) and very different levels of receptor cell surface expression (11-94% wt level) the basal cAMP accumulation determined in transfected COS-7 cells was surprisingly uniform (6.5-8.0 over wt basal). None of the point mutated receptors constitutively activates the phospholipase C cascade. In contrast the deletion 613-621 mutant showed constitutive activity for the IP pathway with a twofold increase in basal IP accumulation compared to the wild type TSHR. All investigated TSHR-mutants showed a TSH-stimulated receptor down-regulation, which seems to be independent of the phospholipase C pathway. CONCLUSIONS: The uniform basal cAMP values in spite of the large variation in specific constitutive activity values suggest that the COS 7 cell overexpression system used for the in vitro characterization is partly regulated. This regulation is most likely due to receptor down regulation. The TSHR deletion mutant (613-621) showed a constitutive activity for both the Galphas and the Galphaq/11 pathways. The TSH-mediated IP-stimulation by this mutant contrasts with its unresponsiveness to TSH for cAMP accumulation and therefore supports the model of different active conformations of the TSHR. PMID- 11012573 TI - Goitre prevalence and thyroid abnormalities at ultrasonography: a comparative epidemiological study in two regions with slightly different iodine status. AB - OBJECTIVE: The association between severe iodine deficiency and endemic goitre is well established, but little information is available on the relation between milder degrees of iodine deficiency and goitre prevalence. SUBJECTS: In a comparative epidemiological study performed in two regions in Denmark, we examined 4649 subjects from the general population, women aged 18-65 years and men aged 60-65 years. METHODS: Ultrasonography and palpation of the thyroid was performed in all participants. Iodine excretion was measured in casual urine samples. Previous thyroid disease was detected by questionnaires, personal interviews and tracing of records. RESULTS: The median iodine excretion was 61 microg/l (mild iodine deficiency (ID)) and 45 microg/l (moderate ID) in the two regions. Median thyroid volume at ultrasonography was 11. 9 ml (mild ID) and 13.6 ml (moderate ID), P <0.001, and thyroid enlargement was found in 15.0% (mild ID) and 22.6% (moderate ID), P<0.001. Goitre prevalence increased in both regions with age to the age group 40-45 years, but not after that age. Subjects who had moved from the moderate ID to the mild ID area had the same prevalence of thyroid enlargement as the subjects staying permanently in the mild ID area. Thyroid nodules at ultrasonography were found in 30% in both regions, but nodules were larger and more often palpable in the moderate ID area. Palpable goitre was found in 9.8% (mild ID) and 14.6% (moderate ID), P<0.001. The greatest regional difference in thyroid abnormalities was found among men. CONCLUSION: Marked differences in the prevalence of thyroid abnormalities were found in these regions with modest differences in iodine excretion. PMID- 11012574 TI - Postpartum thyroid dysfunction in women with normal thyroid function during pregnancy. AB - OBJECTIVE: The aim of this study was to establish the risk of postpartum thyroid dysfunction (PPTD) in women who had normal thyroid function during pregnancy and no history of thyroid disease. DESIGN: Four thousand and twenty-two consecutive pregnant women were screened for thyroid function and antithyroid antibody. Among women with normal thyroid function during pregnancy and no history of thyroid disease, thyroid function were assessed in 131 of 388 antithyroid antibody positive (Group I) and 1030 of 3503 antibody negative (Group II) women at 1 and 3 months postpartum. In Group I women who experienced PPTD, the frequency of later manifestation of Hashimoto's disease was compared according to titres of antithyroid antibodies. MEASUREMENTS: Blood samples in early pregnancy, and at 1 month and 3 months postpartum were obtained using the dried blood spot method. Levels of fT4 were measured by RIA, TSH by fluoroimmunoassay or ELISA, antimicrosome antibody (AMC) and antithyroglobulin antibody (ATG) by indirect agglutination reactions. RESULTS: The prevalence of PPTD at 1 month and 3 months postpartum were found to be 6.9% and 21.3% in Group I, and 5.3% and 4.7% in Group II, respectively. The prevalence of PPTD was significantly higher at 3 months postpartum in Group I (P<0.05). 27.3% of women with PPTD in Group I were later found to have Hashimoto's disease and 9.1% manifested hypothyroidism without goitre. A high AMC titre (> or = 25600) at 3 months postpartum in women with PPTD was related to the manifestation of Hashimoto's disease. AMC titres of PPTD women and women who developed Hashimoto's disease were significantly higher than those of control women who did not experience PPTD. CONCLUSION: A high prevalence of PPTD was found in women with antithyroid antibodies who were euthyroid during pregnancy. Prolonged follow-up of the subsequent thyroid function may be needed in women who experience PPTD and/or show a high titre of antithyroid antibody. PMID- 11012575 TI - How common are polycystic ovaries and the polycystic ovarian syndrome in women with Cushing's syndrome? AB - OBJECTIVE: Women with Cushing's syndrome (CS) may present with menstrual irregularity and symptoms/signs of hyperandrogenism, a phenotype similar to that of the polycystic ovarian syndrome (PCOS); however, currently there are no data on the prevalence of either polycystic ovaries (PCO) and/or PCOS in patients with CS. The aim of this study was to investigate their presence among women of reproductive age presenting with CS by analysing clinical, endocrinological and ultrasonographic features. DESIGN: Prospective study of all women within the reproductive age (range 18-40 years) who presented with CS between August 1994 and January 2000. SUBJECTS AND METHODS: Thirteen women (median age 32 years, range 18-39 years) with CS were evaluated. The diagnosis of CS was based on the presence of appropriate clinical features and an elevated serum midnight cortisol with failure to suppress 0900 hours serum cortisol to less than 50 nmol/l following a formal low-dose dexamethasone suppression test (LDDST). All women had their clinical features relevant to possible hyperandrogenism, menstrual disorder and infertility recorded, and circulating gonadotrophins, oestradiol, androgens and SHBG levels measured; ovarian ultrasonography was performed during their initial assessment. Relevant MR/CT imaging of the pituitary and/or adrenal glands was performed. RESULTS: Eleven women had ACTH-dependent CS [nine Cushing's disease (CD), one ectopic ACTH syndrome due to a bronchial carcinoid, one periodic CS of unknown origin); two patients had ACTH-independent CS (adrenal adenomas). All women with CS had at least one symptom/sign of hyperandrogenism (13 hirsutism, seven acne, five male-pattern alopecia). Nine women (70%) had menstrual disturbances (four oligomenorrhoea, four amenorrhoea, one polymenorrhoea) while four women (30%) had a normal menstrual pattern. Serum oestradiol levels for the group as a whole were similar to those observed in the early follicular phase of normally menstruating women; however, seven women had low oestradiol, LH and FSH levels suggestive of hypogonadotrophic hypogonadism. Serum androgen levels (testosterone, androstendione and DHEAS), even in the presence of symptoms/signs of hyperandrogenism, were within the normal reference range but SHBG levels were uniformly decreased even in women with normal menstrual cycles. There was a negative correlation between urinary free cortisol, but not mean serum cortisol, and serum oestradiol, testosterone and SHBG levels (r = - 0.8, r = - 0.86 and r = - 0.66, P<0.02, P<0.01 and P<0.05, respectively), but not LH or FSH levels. Despite the fact that seven of these 13 patients lacked normal gonadotrophin stimulation, ovarian volumes of both ovaries were relatively preserved: right 7.3 ml, range 2.8-12.8 ml, and left 5.3 ml, range 2.3-13 ml. Women who were defined as oestrogen sufficient (E2 > 140 pmol/l) had higher serum androstenedione, and lower urinary free cortisol levels, than women who were oestrogen deficient (E2 < 140 pmol/l). Six of the 13 women (46%) had ovarian morphology suggestive of PCO, four of six oestrogen sufficient women and two of seven oestrogen deficient women. The results did not differ according to the underlying cause of CS. CONCLUSIONS: PCO and PCOS are common in women with Cushing's syndrome; women with Cushing's syndrome and only moderately elevated cortisol secretion maintain gonadotrophin stimulation to the ovary with normal oestradiol levels, in contrast to women with Cushing's syndrome and higher cortisol secretion who develop hypogonadotrophic hypogonadism. However, even in the latter group, high ovarian volumes were maintained and some had ovarian morphology suggestive of PCO. PMID- 11012576 TI - The prevalence of polycystic ovaries in women with a history of gestational diabetes. AB - OBJECTIVE: Women with a history of gestational diabetes mellitus (GDM) and women with polycystic ovary syndrome (PCOS) both demonstrate abnormalities in insulin action and secretion, and both are at increased risk of developing type 2 diabetes. To determine whether these similarities reflect a common pathophysiological basis, we examined the prevalence of ultrasound-based polycystic ovarian morphology in a large multiethnic group of women with a history of GDM and a group of women who had normal glucose tolerance during pregnancy. PATIENTS AND DESIGN: We studied 91 women with previous GDM (48 European, 20 South Asian, 10 Afro-Caribbean and 13 of other or mixed ethnicity) and 73 normoglycaemic control women (56 European, one South Asian, 14 Afro Caribbean and two of other or mixed ethnicity), a median (interquartile range) of 20 (11-36) and 29 (17-49) months postpartum, respectively. A detailed history was taken, and the prevalence of PCO morphology on ultrasound scan was assessed. Fasting lipids, insulin, glucose status, gonadotrophins and testosterone were measured. Estimates of beta-cell function (%B) and insulin sensitivity (%S) were derived using the HOMA algorithm. RESULTS: Women with previous GDM had higher fasting glucose (5.4 (4. 8-6.0) vs. 4.7 (4.4-5.0) mmol/l, P<0.0001) and features reminiscent of syndrome X: higher BMI (26.4 (22.8-31.4) vs. 23.8 (21. 0-27.5) kg/m2, P = 0.002), waist/hip ratio (0.82 (0.79-0.88) vs. 0. 77 (0.73-0.81), P<0.0001), fasting insulin (165 (68-299) vs. 54 (24-156) pmol/l, P<0.0001), triglycerides (1.1 (0.8-1.6) vs. 0.8 (0.6-1.1) mmol/l, P<0.0001) and lower insulin sensitivity (%S) (27 (16-62) vs. 86 (34-139)%, P<0.0001) compared to control women. The prevalence of PCO was higher in the previous GDM group than in the control subjects (47/91 (52%) vs. 20/73 (27%), chi2 = 9.86, P = 0. 002 overall, odds ratio 2.7, P = 0.007 by logistic regression allowing for ethnicity). There was no difference in any metabolic parameter between the post GDM PCO group and the post-GDM normal ovaries group, but irregular cycles were more prevalent in the PCO group (22/47 (47%) vs. 9/44 (21%), chi2 = 7.03, P = 0.008). CONCLUSIONS: We found a higher prevalence of polycystic ovarian morphology in women with a history of gestational diabetes. Among the women with previous gestational diabetes, irregular cycles were more prevalent in the PCO group than in the women with normal ovarian morphology, but no other differences in endocrine or metabolic parameters were detected. These findings confirm an association between PCO and gestational diabetes and suggest that women with gestational diabetes display metabolic abnormalities irrespective of ovarian morphology. PMID- 11012577 TI - Growth and endocrine disorders in multisystem Langerhans' cell histiocytosis. AB - INTRODUCTION: Langerhans' cell histiocytosis is a rare disorder, with diabetes insipidus occurring in up to half of patients. Causes of growth failure include the illness itself, treatments used and growth hormone insufficiency. PATIENTS AND METHODS: We identified all patients with an endocrinopathy secondary to Langerhans' cell histiocytosis (LCH). Growth data were analysed from all patients with multisystem involvement. RESULTS: Of 144 patients with multisystem LCH, 50 had an endocrinopathy, 49 of whom had diabetes insipidus. Growth hormone insufficiency (GHI) was present in 21 patients, seven of whom had other anterior pituitary deficiencies as well (gonadotrophin deficiency + GHI n = 2, gonadotrophin deficiency + TSH deficiency + GHI n = 2, panhypopituitarism n = 3). GH insufficiency, the development of which appeared to be independent of pituitary radiation, occurred at a median age of 8.3 years (4.7-18 years) and at a median interval of 3.5 years (0-11.8 years) after diagnosis of LCH. The median height SDS at diagnosis of growth hormone insufficiency was -2.9. Thirteen of the patients with growth hormone insufficiency attained final height with a median height SDS of -1.2. The final height SDS of 15 patients without GH insufficiency was closer to target height SDS, but not statistically different from that of the GH insufficient group. CONCLUSIONS: GH therapy significantly improves growth in GH insufficient patients with Langerhans' cell histiocytosis. Early institution of GH therapy may further improve height outcome. However, most children with Langerhans' cell histiocytosis regardless of endocrine function, failed to reach target height. PMID- 11012578 TI - Androgens and penile erection: evidence for a direct relationship between free testosterone and cavernous vasodilation in men with erectile dysfunction. AB - OBJECTIVE: Androgens are essential in the maintenance of nitric oxide-mediated erectile activity in the rat. The objective of the present study was to investigate the role of androgens in regulating trabecular smooth muscle relaxation in the corpus cavernosum in response to vasoactive challenge in men with erectile dysfunction (ED). DESIGN: Retrospective, double-blind correlation analyses. PATIENTS: Fifty-two impotent patients without confounding risk factors for ED were obtained from a total of 250 undergoing diagnostic evaluation. MEASUREMENTS: All patients had dynamic colour duplex ultrasound (D-CDU) and hormonal evaluation for LH, total and free testosterone, SHBG and oestradiol. RESULTS: Based upon D-CDU results patients were diagnosed as having arteriogenic (AR, n = 18; mean age 51) or corporeal venocclusive (CVO, n = 13; mean age 49) ED; in other patients (n = 21, mean age 43) a diagnosis of psychogenic (P)-ED was made by comprehensive psychogenic testing and confirmed by normal D-CDU results. AR and CVO patients had altered compliance of cavernous arteries recorded by D CDU [20-25% lower resistive index (RI) than patients with psychogenic ED], and lower free testosterone (FT) levels than psychogenic patients [42.3 +/-3.5 SE and 49.3+/-5.2 vs. 75.2+/-7.6 pmol/l, respectively; P<0.01]. More important, in all patients there was a strong direct correlation between resistive index values and FT levels (r = 0.47, P = 0.002); the relationship was maintained also when adjusted for age, SHBG and oestradiol (r = 0.37, P = 0.02). CONCLUSIONS: These results indicate that in men with erectile dysfunction low free testosterone may correlate independently of age with the impaired relaxation of cavernous endothelial and corporeal smooth muscle cells to a vasoactive challenge. These findings give clinical support to the experimental knowledge of the importance of androgens in regulating smooth muscle function in the penis. PMID- 11012579 TI - Adrenal hyperandrogenism in adolescent girls with a history of low birthweight and precocious pubarche. AB - OBJECTIVE: Girls with precocious pubarche (PP) are at increased risk for ovarian dysfunction, hyperinsulinism and dyslipidaemia in adolescence, in particular when PP is preceded by reduced fetal growth. However, it is not known whether PP girls still have adrenal hyperandrogenism after puberty and if so, which fraction of PP girls develops so-called functional adrenal hyperandrogenism (FAH), an entity characterized by ACTH-dependent 17-ketosteroid excess. PATIENTS AND DESIGN: Data were longitudinally collected from 47 girls with PP: at birth (weight for gestational age), at diagnosis of PP (age 6.7+/- 1.1 years) and in adolescence (age 15.0+/-1.9 years). MEASUREMENTS: Serum dehydroepiandrosterone sulphate (DHEAS) and androstenedione were measured at PP diagnosis, as well as the 17 hydroxyprogesterone (17-OHP) response to ACTH; postpubertal evaluation included assessment of adrenal and ovarian function, and of insulin responses to a glucose load. PP girls were considered to have FAH in adolescence if both DHEA and androstenedione responses to ACTH were excessive (> 1500 ng/dl and > 350 ng/dl, respectively). RESULTS: At diagnosis of PP, girls had high DHEAS and androstenedione levels, as well as high 17-OHP responses to ACTH. In adolescence, PP girls had a normal BMI, presented with mild hirsutism and had high baseline and post-ACTH concentrations of most adrenal androgens, low SHBG levels and tended to have hyperinsulinemia and to present biological signs of ovarian hyperandrogenism. More than a third of the PP cohort developed FAH in adolescence. Neither baseline DHEAS, androstenedione, nor post-ACTH 17-OHP values at diagnosis of PP predicted the development of FAH in adolescence. In PP girls, only a low weight at birth was found to be significantly associated with subsequent FAH. CONCLUSIONS: These longitudinal findings in girls with PP point to the possibility of an endocrine sequence of prenatal onset: low weight at birth, PP in childhood and adrenal hyperandrogenism in adolescence. The pathophysiological mechanisms underpinning this newly recognized sequence remain to be identified. PMID- 11012580 TI - Optic chiasmal herniation--an under recognized complication of dopamine agonist therapy for macroprolactinoma. AB - The initial presentation of macroprolactinoma with visual field impairment, especially in males, is well recognized. Successful treatment with dopamine agonist therapy is characterized by a reduction in hyperprolactinaemia and often rapid and progressive resolution of the visual impairment. A small proportion of patients may subsequently develop a secondary deterioration in both their visual fields and visual acuities despite normalization of prolactin levels and tumour shrinkage. When pituitary apoplexy can be excluded this may result from traction on the optic chiasm which is pulled down into the now partially empty sella. We report a series of seven patients in whom chiasmal traction is believed to be the cause of their secondary deterioration in visual acuity occurring after dopamine agonist therapy for macroprolactinoma. The clinical history of two patients both of whom had rapid resolution of field defect with bromocriptine therapy but subsequently developed a recurrence of their bitemporal hemianopia is detailed. In both patients MRI scanning showed not only tumour involution but also marked optic chiasm herniation into the pituitary fossa. Surgical treatment was considered too risky; but on reduction of bromocriptine dosage the field defect improved in both cases; there was a modest elevation of prolactin and a degree of tumour re-expansion. The latter is believed to have released tethering of the optic chiasm and/or its vascular supply and thus obviated the need for surgery. Regular monitoring of visual fields in patients with macroprolactinoma receiving medical treatment is therefore important. Early recognition of secondary field loss due to chiasmal herniation enables correction of the visual field loss by manipulation of the medical therapy. PMID- 11012581 TI - Gigantism due to growth hormone excess in a boy with optic glioma. AB - True gigantism is rare in early childhood and is usually due to excess GH secretion from a pituitary adenoma. We report a case in which the endocrine abnormality is secondary to an optic glioma. Careful endocrine evaluation has shown that GH peak amplitude was not increased but rather there was failure of GH levels to suppress to baseline and a lack of pulsatility. There is no evidence of a direct secretory role for the tumour and we postulate that the tumour is affecting GH secretion through an effect on somatostatin tone. Specific tumour therapy is not indicated for this patient in the absence of mass effect or visual disturbance. The GH excess is being treated with somatostatin analogue (Octreotide) and as he has developed precocious puberty he is also receiving long acting GnRH analogue (Zoladex). This boy appears likely to have neurofibromatosis type 1 (NF1) which raises the question of subtle GH excess in NF1 patients with tall stature. PMID- 11012582 TI - Cushing's syndrome in a 16 year old girl due to ectopic ACTH precursor production from a pancreatic tumour. PMID- 11012583 TI - Muscle mass and function in thyrotoxic patients before and during medical treatment. PMID- 11012584 TI - Muscle mass and function in thyrotoxic patients before and during medical treatment PMID- 11012585 TI - Management of nail psoriasis. AB - Nail psoriasis is often difficult to cure, but may respond to a range of different approaches used alone or together. As with treatment of psoriasis affecting skin, relapse is common and therapies may need to be maintained or repeated. The exact benefits of many of these treatments are not well documented in the literature. Basic nail care is important and topical therapies represent the main modality of treatment for the majority of cases. In severe nail disease, where there is a hypertrophic element, injection therapy with triamcinolone may be helpful. PUVA and other forms of radiation may be of benefit, but as with systemic therapy, they are usually useful in the context of treatment of psoriasis elsewhere on the body. PMID- 11012586 TI - Cutaneous leishmaniasis. AB - Leishmaniasis is a major World health problem, which is increasing in incidence. In Northern Europe it is seen in travellers returning from endemic areas. The protozoa is transmitted by sandflies and may produce a variety of clinical syndromes varying from a simple ulcer to fatal systemic disease. This review considers the management of simple cutaneous leishmaniasis. Patients usually have a single ulcer which may heal spontaneously, requiring only topical, or no treatment at all. Lesions caused by Leishmania braziliensis may evolve into the mucocutaneous form, 'espundia', and should be treated with systemic antimony. Sodium stiboglucoante 20 mg/kg/day i. v. for 20 days is the appropriate first line treatment in these cases. Although it may cause transient bone marrow suppression, liver damage, a chemical pancreatitis, and disturbances in the electrocardiogram, it appears to be safe. The success of treatment should be assessed 6 weeks after it has been completed and patients should be followed up for 6 months. PMID- 11012587 TI - General practitioner referral guidelines for dermatology: do they improve the quality of referrals? AB - Guidelines have proliferated in recent years in all areas of medicine, sometimes with little evaluation of their effect. Referral guidelines for dermatology were compiled by the dermatologist at the Royal Surrey County Hospital in consultation with local general practitioners. An audit was undertaken to assess how appropriate referrals were just before and after distribution of the guidelines and was repeated 2 years later to determine whether they had made any significant impact. The results revealed a 40% increase in the number of appropriate referrals immediately after introduction of the guidelines, but this was not sustained 2 years later. Five common conditions accounted for two-thirds of inappropriate referrals before and after the guidelines were sent. The need for continued general practitioner education in dermatology to reinforce referral guidelines is demonstrated. PMID- 11012588 TI - Erythema nodosum in Singapore. AB - A retrospective study was carried out of all patients presenting with erythema nodosum over a 3-year period (1994-1997) at the National Skin Centre, Singapore. There were 75 patients, aged 4-70 years with a peak (40%) in the 21-30-year-old age group; 65 were female and 10 were male, giving a female to male ratio of 6.5 : 1. Precipitating factors included viral respiratory tract infections and streptococcal pharyngitis in 20 cases (26%), tuberculosis in two cases (3%), pregnancy in three cases (4%) and miscellaneous conditions: Behcet's syndrome (two cases), gonorrhoea (one case), varicella (one case) and cat-scratch disease (one case). The cause was unknown in 45 cases (60%). The pattern of erythema nodosum in Singapore is similar to that reported in the Western literature except that sarcoidosis and inflammatory bowel disease are uncommon associations. PMID- 11012589 TI - Multiple basal cell carcinomas and malignant melanoma following radiotherapy for ankylosing spondylitis. AB - We present the case of a 53-year-old Caucasian woman with seven basal cell carcinomas and one malignant melanoma in situ along her back overlying her spine, which was irradiated in 1968 for ankylosing spondylitis. These lesions developed between 1997 and 1999. She has no other known risk factors for cutaneous malignancy, in particular no history of excessive sun exposure. She has skin type 2. Molecular studies of glutathione S-transferase and cytochrome P450 status showed her genotype not to constitute an overall increased inherited susceptibility. We therefore postulate that all her skin cancers have arisen as a consequence of her radiotherapy. To our knowledge this is the first case of multiple basal cell carcinoma in addition to a malignant melanoma following radiotherapy for benign disease. PMID- 11012590 TI - Incipient osteomalacia occurring in chronic actinic dermatitis. AB - Incipient osteomalacia developed in a Pakistani patient living in the UK after strict sunlight avoidance forming part of the management of the photosensitivity disorder, chronic actinic dermatitis. The patient's skin type and diet, which included calcium-binding phytates in chappattis, had increased his risk of the condition. Proximal muscle weakness and bony tenderness resulting from the disorder resolved on vitamin D replacement therapy. PMID- 11012591 TI - Chemotherapy-induced acral erythema: report of a case and immunohistochemical findings. AB - Chemotherapy-induced acral erythema (CAE) is an uncommon and distinct reaction seen in patients receiving high-dose chemotherapy. The exact pathogenic mechanisms of this disorder are still unknown. We report a 27-year-old woman who presented with red, swollen and painful macules on both palms, clinically consistent with this disease. Histological examination demonstrated vacuolar degeneration of the basal cell layer and spongiotic blisters in the epidermis, especially in the atrophied eccrine ducts and papillary oedema with mild perivascular infiltration of mononuclear and hypersegmented neutrophils. Immunohistochemistry showed that the infiltrating mononuclear cells were CD3 CD16+CD56+ leucocyte function antigen-1+, possibly natural killer cells. The eccrine ducts expressed HLA-DR and intracellular adhesion molecule-1 (ICAM-1). Our findings suggest that cell-to-cell interaction between NK cells and keratinocytes in the eccrine apparatus may induce CAE and may be involved in the pathogenesis of the skin reaction in our patient and possibly in this disease. PMID- 11012592 TI - Calciphylaxis in the absence of renal failure. AB - We report a patient with severe bilateral leg ulceration that was resistant to treatment. A biopsy confirmed the cause as calciphylaxis. Calciphylaxis refers to a syndrome of calcium deposition in the small and intermediate dermal vasculature which can lead to epidermal ischaemia, ulceration and necrosis. Most cases occur in those with chronic renal failure and secondary hyperparathyroidism. We describe the rare presentation of calciphylaxis in a patient with normal renal function and primary hyperparathyroidism who had many classical features. Unfortunately she developed gangrene, sepsis and died. PMID- 11012593 TI - Orificial tuberculosis and Kaposi's sarcoma in an HIV-negative individual. AB - Orificial tuberculosis is a rare form of cutaneous tuberculosis. It happens mainly in immunocompromised patients and is characterized by rapidly ulcerating papules in the orifices, including oropharynx or anus, due to autoinoculation of internal tuberculosis. It is a symptom of advanced underlying disease. Kaposi's sarcoma occurs more frequently with the increase of HIV-infected patients. We report on an HIV-negative individual who developed rapid-deteriorating orificial tuberculosis concomitant with Kaposi's sarcoma and had a fatal outcome. PMID- 11012594 TI - Dermatitis herpetiformis evolving into bullous pemphigoid: a probable example of 'epitope spreading'. AB - We report a case of dermatitis herpetiformis which, 11 years after its original diagnosis, evolved into bullous pemphigoid. Only a few similar cases supported by immunofluorescence studies have been reported previously, and we believe that they represent examples of 'epitope spreading', an increasingly recognized phenomenon used to explain the coexistence of autoimmune diseases. PMID- 11012595 TI - Co-existing actinic granuloma and giant molluscum contagiosum. AB - Actinic granuloma of O'Brien and giant molluscum contagiosum in immunocompetent adults are rare skin disorders. There have been no previous reports of these two conditions occurring together. We report this occurrence in a 57-year-old Caucasian woman. PMID- 11012596 TI - Persistent annular erythema of infancy associated with intestinal Candida colonization. AB - We report a case of persistent annular erythema of infancy in a 4-month-old boy. Physical and laboratory parameters showed no sign of internal disease or specific infection except a massive Candida albicans colonization (> 103 organisms/mm3) of the lower gastrointestinal tract. Oral treatment with amphotericin B for 2 weeks resulted in a complete remission of the skin lesions indicating Candida colonization as a trigger. PMID- 11012597 TI - An unusual case of purpuric erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is the most common of the erythropoietic porphyrias. Recent advances in diagnostic laboratory tests have led to the discovery of a number of previously undiagnosed cases. We describe a case of EPP presenting late with a purpuric eruption and discuss the pathogenesis and significance of purpura in EPP. PMID- 11012598 TI - Rupioid psoriasis with arthropathy. AB - A 31-year-old male presented with limpet-like, cone-shaped skin lesions on the scalp, the extremities, and the trunk of 4 months' duration, and had had severe joint pain in his right fingers, wrist, and knee for 1 month. Radiological examination revealed arthritis of the above mentioned multiple joints. Rheumatoid factor was serologically negative. Histopathological findings showed dense inflammatory cell infiltration and remarkable Munro's microabscesses in the horny layer in addition to psoriasiform epidermal hyperplasia, and predominant dermal oedema. Based on clinicopathological findings, a diagnosis of rupioid psoriasis with arthropathy was made. Following treatment for arthralgia using low doses of systemic steroid, the effects of cyclosporin combined with topical steroids was seen to alleviate dramatically the skin lesions and arthritis within 2 weeks. There was no recurrence of such skin and joint lesions during a follow-up period of 1 year. PMID- 11012600 TI - Laser therapy of telangiectatic leg veins: clinical evaluation of the 810 nm diode laser. AB - Telangiectatic leg veins (TLV) are experienced by millions of women and men. As they cause significant cosmetic embarrassment, many women seek effective treatment. Options include sclerotherapy and treatment with different lasers. We investigated the safety and efficacy of an 810-nm diode laser in the treatment of TLV. Ten females had treatment to a >/= 25 cm2 area of TLV four times at 4-weekly intervals at 25-35 W, 0.1 s duration, 0.5 s interval and a 5-mm circular spot size (fluence of 12.7-17.8 J/cm2). Assessment of response was performed 4 weeks after the final treatment and patients were asked to complete a questionnaire requesting information regarding the treatment. No statistically significant clearing of the leg veins was demonstrated although some improvement was noted in individual patients. Side-effects were negligible, consisting of discomfort or mild pain. There was no scarring, purpura or pigment alteration. A visual analogue scale questionnaire (0-10) revealed that patients found the treatment acceptable (range, 4-10). Overall difference as assessed by the patients ranged from 0 to 8.4 and degree of satisfaction with the treatment from 0 to 9.8. Currently, the treatment of choice for TLV remains sclerotherapy. Our results suggest that the 810-nm diode laser is safe and well tolerated. Further research could establish whether effectiveness can be improved with higher fluences or in combination with sclerotherapy. PMID- 11012599 TI - Langerhans cell migration. AB - Epidermal Langerhans cells (LC) play pivotal roles in the induction of cutaneous immune responses. Encounter with antigen in the skin, or other stimuli, cause the mobilization of LC and their migration, via afferent lymphatics, to draining lymph nodes where they localize within the paracortex. During their movement from the skin LC acquire the characteristics of immunostimulatory dendritic cells (DC) such that the antigen-bearing cells which accumulate in lymph nodes are able to provoke specific T-lymphocyte responses. Epidermal cytokines initiate and regulate LC migration (and maturation), of particular importance being interleukin-1beta and tumour necrosis factor-alpha. Collectively, these cytokines, together with relevant chemokine receptor-ligand interactions, effect the liberation of LC from the epidermis and their directed movement to, and localization within, peripheral lymph nodes. Described here are the phenotypic changes induced during the activation of LC and the mechanisms through which their migration is regulated. PMID- 11012601 TI - Collagenase digestion and mechanical disaggregation as a method to extract and immunophenotype tumour lymphocytes in cutaneous T-cell lymphomas. AB - Various enzymatic or mechanical methods have been proposed in the past to dissociate cells from different solid tissues. An automated mechanical disaggregation device (Medimachine) has recently been proposed. Unfortunately, most of these techniques are associated with a high cellular damage and a low cell recovery and are difficult to apply to skin biopsies. In this paper, we propose a combined enzymatic and mechanical method based on Medimachine, useful for the isolation of skin infiltrating T-lymphocytes from small cutaneous biopsies. As this method is easy and allows for a more correct qualitative and quantitative cytofluorimetric analysis of the lymphocyte subsets, it may be useful in the immunophenotyping of cutaneous T-cell lymphomas. PMID- 11012602 TI - Do plasminogen activators play a role in lichen sclerosus? AB - The histological changes of lichen sclerosus suggest that significant remodelling of the extracellular matrix is occurring. As the proteases of the plasminogen activator system have been implicated in tissue remodelling, cell migration and tumour invasion, we performed an immunohistochemical study to look for evidence of alteration in the expression of plasminogen/plasmin, urokinase-type plasminogen activator, tissue-type plasminogen activator and alpha2-antiplasmin in biopsies of clinically typical vulval lichen sclerosus obtained from 11 untreated adult women. Normal vulva obtained from gynaecological procedures and samples of the patients' uninvolved thigh tissue were used as controls. No significant difference was seen in the staining pattern between the lichen sclerosus tissue and control tissue. However, although we found no immunohistochemical evidence that the plasminogen activator system is involved in the pathogenesis of vulval lichen sclerosus, it may be that other proteases are involved. PMID- 11012603 TI - Disaccharide analysis of skin glycosaminoglycans in atrophoderma of Pasini and Pierini. AB - There are divergent opinions as to whether atrophoderma of Pasini and Pierini (APP) is a nosologic entity or a primary atrophic morphoea. In this study, we used high performance liquid chromatography to analyse the skin disaccharide contents of glycosaminoglycan (GAG) in two patients with APP and compared the results with those from a typical atrophic morphoea patient. Perilesional uninvolved skin was used as a control in each patient. In the atrophic phase morphoea, both the total amount of disaccharide per skin punch-biopsy and the amount of DeltaDi-4S(DS) - the main disaccharide unit of dermatan sulphate - per mg dry weight were increased. These changes were consistent with sclerotic phase morphoea. In contrast, the total amount of disaccharide per skin punch-biopsy was decreased and the amount of DeltaDi-4S(DS) per mg dry weight was decreased or unchanged in APP. Our results suggest that GAG metabolism in APP may be unique and quite different from that in morphoea. PMID- 11012604 TI - Heterozygous germline missense mutation in the p63 gene underlying EEC syndrome. AB - Mutations in the p63 gene have recently been delineated as the molecular basis for some cases of the ectrodactyly, ectodermal dysplasia and cleft lip/palate (EEC) syndrome, an autosomal dominant disorder (MIM 129900). In this report, we describe a 35-year-old woman with EEC syndrome and document a heterozygous germline missense mutation, R304W, in exon 8 of the p63 gene. As with most other p63 mutations in EEC syndrome, this mutation has arisen de novo and is located within the core DNA-binding domain of p63. Identification of this mutation has implications for genetic counselling and the feasibility of future DNA-based prenatal diagnosis in this individual. PMID- 11012605 TI - Dowling Oration 2000. Dermatology patients: what do they really need? AB - Dermatology services must be organized to meet the needs of patients. These needs change as society changes and medical knowledge increases. Considerable insight into the true needs of individual and groups of patients can be gained from the use of quality of life measures. Patients with widespread inflammatory skin disease are most severely handicapped by their skin disease and can be most helped by dermatology services. It is essential that such patients be given priority in the delivery of dermatological care. Where necessary, protected clinic time and specialist support services should be created to ensure that such patients are not adversely affected by pressures to review patients in other diagnostic groups. PMID- 11012606 TI - PUVA therapy of granuloma annulare. PMID- 11012607 TI - Are patients with localized nodular granuloma annulare more likely to have diabetes mellitus? PMID- 11012608 TI - Pemphigoid gestationis with intra-uterine death associated with foetal cerebral haemorrhage in the mid-trimester. PMID- 11012610 TI - Serological markers of disease activity in tuberculosis and HIV infection. PMID- 11012611 TI - Pulmonary tuberculosis and serum IgE. AB - Several recent studies indicate that mycobacterium or viral infection may reduce IgE levels or suppress atopy or both. The present study was undertaken to investigate whether Mycobacterium tuberculosis infection and its successful treatment down-regulate serum total IgE levels, a marker of a Th2 response, due to enhancement of a Th1 response in adult patients with tuberculosis (TB). We prospectively studied the changes in serum total IgE and DTH response to tuberculin, a marker of a Th1 response in 10 healthy controls, 20 patients with pulmonary TB, and 19 asthma patients without TB. Measurement of serum total IgE and tuberculin skin tests were performed before initiation of treatment and after successful completion of 6 months treatment in TB patients, and at the corresponding intervals in controls and asthmatics. The initial serum total IgE concentrations were significantly higher in TB patients than in healthy controls (282 +/- 26 U/ml (mean +/- s.e.m.) in TB patients versus 126 +/- 56 U/ml in controls; P = 0.03). However, serum total IgE concentrations significantly decreased (282 +/- 26 U/ml before versus 151 +/- 12 U/ml after treatment; P = 0.03) and tuberculin indurations significantly increased (23.6 +/- 1.8 mm before versus 29.6 +/- 2.1 mm after treatment; P = 0.04) in TB patients. In contrast, initial serum IgE concentrations and tuberculin indurations did not differ significantly from post-observation data in both healthy controls and asthmatics (P>0.30). The present study confirmed that immune responses to M. tuberculosis down-regulate a Th2 immune response, and might contribute to the decreased prevalence of allergic disorders. PMID- 11012609 TI - Chronic granulomatous disease. PMID- 11012612 TI - Atopic eczema is associated with delayed maturation of the antibody response to pneumococcal vaccine. AB - The aim of this study was to investigate a previously undocumented observation, that children with atopic eczema under 9 years of age tended to have a poor antibody response to Pneumococcal vaccination. Thirty-five children (mean age 8.8 years, range 3-16 years) with moderate to severe atopic eczema but no history of systemic infection were studied retrospectively. Pneumococcal antibody responses after immunization with Pneumovax II were compared with a hospital control group consisting of 36 children (mean age 6.0 years, range 3-16 years) with recurrent upper respiratory tract infections. Only 17% of children with atopic eczema aged 3-8 years responded to Pneumovax. This response was significantly poorer than that of the controls (57%) (odds ratio 0.20, 95% confidence interval (CI) 0.05 0.84, P = 0.03). There were no significant differences in the levels of total IgG2, the component of IgG associated with protective antibody responses to Pneumococcus between the two groups. Delay in maturation of the total IgG and IgG2 antibody response to Pneumococcus is a feature in this group of children with moderately severe atopic eczema. PMID- 11012613 TI - Role of caspases in dexamethasone-induced apoptosis and activation of c-Jun NH2 terminal kinase and p38 mitogen-activated protein kinase in human eosinophils. AB - Eosinophils are the principal effector cells for the pathogenesis of allergic inflammation. Glucocorticoids such as dexamethasone have long been used therapeutically for eosinophilia in allergic inflammation by inducing eosinophil apoptosis, but little is known about the intracellular mechanisms mediating dexamethasone-induced apoptosis. In the present study, we investigated the effect of dexamethasone on three mitogen-activated protein kinases (MAPK) involved in the intracellular signalling pathway: c-Jun NH2-terminal kinase (JNK), p38 MAPK and extracellular signal-regulated kinase (ERK). We found that dexamethasone could activate JNK and p38 MAPK in a time-dependent manner but not ERK. Further, SB 203580, a specific p38 MAPK inhibitor, was additive with dexamethasone in inducing eosinophil apoptosis, while JNK1/2 antisense phosphorothioate oligodeoxynucleotides did not show any significant effect. These suggest that dexamethasone-induced JNK1/2 and p38 MAPK activation are not crucial to the induction of apoptosis. Pretreatment of eosinophils with benzyloxycarbonyl-Val Ala-Asp-fluoromethylketone (Z-VAD.FMK), a broad-spectrum caspase inhibitor, could inhibit dexamethasone-induced apoptosis in eosinophils dose-dependently. Moreover, Z-VAD.FMK partially inhibited dexamethasone-activated JNK and p38 MAPK activities. However, dexamethasone treatment did not activate specific caspase-3, -8 activity in eosinophils compared with spontaneous apoptosis. We therefore conclude that dexamethasone-induced apoptosis and activation of JNK and p38 MAPK activity in eosinophils are regulated by caspases but not through the common apoptosis-related caspase-3, -8 as in other cell types. Elucidation of the important role of caspases in eosinophil apoptosis may facilitate the development of more specific and effective treatment for allergic inflammation. PMID- 11012614 TI - Effects of IL-4 and IL-12 on experimental immune-mediated blepharoconjunctivitis in Brown Norway rats. AB - IL-12 and IL-4 are critical cytokines for Th1 and Th2 differentiation, respectively. To assess the roles of these cytokines in the development of experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats, their effects were tested either in vitro or in vivo. Draining lymph node cells from rats immunized with ragweed pollen (RW) in Al(OH)3 were collected and cultured for 3 days with RW in the presence of IL-4, IL-12, or PBS as a control. After harvesting the culture supernatants for cytokine ELISA and the cells for cytokine reverse transcriptase-polymerase chain reaction, 10 million cells were injected intravenously into syngeneic recipient rats (n = 12 per group). The rats were challenged with RW by eye drops 4 days after transfer. Eyes were harvested for histology 24 h later. Furthermore, IL-12 (500 ng per injection) or PBS was injected intraperitoneally every other day seven times from the day of active immunization (n = 6 per group). One day after the last injection, rats were challenged and EC was evaluated as above. Transfer of cells with IL-4 in vitro augmented eosinophilic infiltration in the conjunctiva compared with the other two groups, whereas IL-12 in vitro suppressed eosinophilic infiltration and increased lymphocytic infiltration. Interferon-gamma production was augmented by IL-12. IL-4 RNA expression was augmented by IL-4. IL-12 administration in vivo augmented lymphocytic infiltration in the conjunctiva without affecting infiltration of eosinophils. In conclusion, IL-4 and IL-12 either in vitro or in vivo augmented Th2 and Th1 immunity, respectively, thus leading to distinct histological features of EC. PMID- 11012615 TI - Eosinophil granule-derived major basic protein induces IL-8 expression in human intestinal myofibroblasts. AB - Eosinophil infiltration occurs in a variety of allergic and inflammatory diseases. The release of preformed mediators from eosinophils may contribute to inflammatory responses. We investigated the ability of eosinophil-derived major basic protein and eosinophil-derived neurotoxin to stimulate production of IL-8 from intestinal myofibroblasts. Intestinal myofibroblasts (18-Co cells) were incubated with major basic protein, eosinophil-derived neurotoxin, or a synthetic analogue of major basic protein, poly-L-arginine. Immunoreactive IL-8 was measured by ELISA and IL-8 mRNA levels were analysed by Northern blot or reverse transcription-polymerase chain assay. Major basic protein induced IL-8 mRNA production and release of significant levels of IL-8 immunoreactive protein. By contrast, eosinophil-derived neurotoxin stimulated little IL-8 release. The induction of IL-8 mRNA by poly-L-arginine was significantly inhibited by actinomycin D. These findings demonstrate a novel interaction between eosinophils and intestinal fibroblasts that may be involved in the pathogenesis of diseases associated with tissue eosinophilia. PMID- 11012616 TI - Antigen-induced IL-17 response in the peripheral blood mononuclear cells (PBMC) of healthy controls. AB - IL-17 is a T cell cytokine with a complex and important role in the immune system. It has been detected in rheumatoid arthritis (RA) synovial membrane and found to stimulate the production of the proinflammatory cytokines IL-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony stimulating factor (GM-CSF) in vitro. To date, there are few data available on the agents that stimulate IL-17 production. We therefore investigated the in vitro IL-17 response to a variety of mitogens and antigens, and compared the IL 17 response to interferon-gamma (IFN-gamma), IL-4, IL-10 and TNF-alpha. In this study we used a type-0 antigen, tetanus toxoid (TT), a type-1 antigen, PPD from Mycobacterium tuberculosis, a potential type-2 rye grass (RG) antigen (Lol I) and an autoantigen SS.B (La), to stimulate PBMC from healthy controls. Cytokine mRNA was measured using semiquantitative reverse transcriptase-polymerase chain reaction and cytokine protein measured using specific ELISA techniques, while the frequency of IL-17-producing T cells was determined by flow cytometry. The mitogens concanavalin A, phytohaemagglutinin and phorbol myristate acetate/ionomycin induced a significant increase in IL-17, with the highest levels being produced by anti-CD3/anti-CD28 stimulation. The antigens TT and PPD significantly increased IL-17 mRNA expression over time, but failed to have such an effect at the protein level. IL-17 protein was also detectable in both antigen specific (TT, SS. B) and non-specific T cell clones, but at levels lower than IFN gamma. IL-17 production did not correlate with either the type-1 cytokine IFN gamma or TNF-alpha or the type-2 cytokine IL-4 or IL-10 at either the mRNA or protein level. PMID- 11012617 TI - Glycosylphosphatidylinositol (GPI)-deficient Jurkat T cells as a model to study functions of GPI-anchored proteins. AB - Many cell surface proteins attached to the membrane by GPI are involved in cell signalling. However, the role of the GPI membrane anchor itself remains poorly understood. GPI-defective cells from patients with paroxysmal nocturnal haemoglobinuria (PNH) are relatively resistant to apoptosis induction. We developed a Jurkat T cell model for GPI deficiency by isolating a GPI-negative mutant, which is defective in the GPI biosynthetic gene PIG-A. Using retroviral PIG-A gene transfer along with the transfer of a vector control, we obtained two genetically identical cell lines, distinguished only by expression of the PIG-A gene and, thus, their ability to produce GPI. Cell proliferation and survival were not affected by this difference. Apoptotic stimuli such as serum starvation and camptothecin exposure elicited similar responses. In contrast, GPI-defective Jurkat cells were more susceptible to Fas-mediated apoptosis than GPI-positive cells. These results indicate that a deficiency in GPI-anchored proteins, as is found in PNH, does not confer resistance to apoptosis. PMID- 11012618 TI - Antibodies, directed towards Campylobacter jejuni antigens, in sera from poultry abattoir workers. AB - Occupational exposure of susceptible humans to Campylobacter jejuni appears to result in resistance to disease. This is believed to be due to acquired protective immunity. To support this hypothesis the levels of C. jejuni-specific IgG and IgM antibodies were determined in sera from poultry abattoir workers. Such individuals are persistently exposed to C. jejuni, but apparently rarely acquire campylobacteriosis. Sera from 43 short-term workers (employed < or = 1 month), 78 long-term workers and 40 blood donors were investigated by ELISA. In 51 individuals a second serum sample, taken at least 1 month after the first, was also investigated. Eight workers had C. jejuni-positive faecal cultures and only one, a short-term worker, had symptoms of campylobacteriosis. There were significantly higher levels of specific IgG antibodies in long-term workers than in either of the other groups. There was no significant difference detectable in specific IgM antibody levels between any of the groups. The results provide supporting evidence that long-term exposure to C. jejuni induces circulating antibodies which reflect apparent reduced susceptibility to disease. Western blotting showed flagellin and polypeptides of 45, 40, 32 and 30 kD bound antibodies significantly more frequently by sera from long-term workers than short-term workers and blood donors. The most commonly detected antigens were the 40-kD (80%) and flagellin (55%). The results indicate that specific serum IgG responses induced by endemic exposure to C. jejuni might be directed towards a small number of protein antigens with apparently conserved epitopes. PMID- 11012619 TI - Induction of necrosis and apoptosis of neutrophil granulocytes by Streptococcus pneumoniae. AB - Apoptosis followed by macrophage phagocytosis is the principal mechanism by which neutrophil granulocytes (PMN) are removed from the site of inflammation. To investigate whether Streptococcus pneumoniae causes apoptosis of PMN, we exposed PMN to viable and heat-killed pneumococci and purified pneumococcal cell walls (PCW). The occurrence of PMN cell death was quantified by flow cytometry using annexin V/propidium iodide labelling of the cells. Intracellular histone associated DNA fragments were quantified by ELISA. The presence of apoptosis was confirmed by in situ tailing. Exposure of PMN to viable pneumococci caused necrosis of the cells. The pneumococcal cytotoxin pneumolysin, the bacterial production of hydrogen peroxide, and PCW contributed to necrosis. Heat-killed pneumococci accelerated the process of apoptosis observed in cultivated non stimulated PMN in vitro. These results demonstrated that pneumococci induce PMN cell death. Depending on the intensity of the stimulus, PMN necrosis and apoptosis were observed. PMID- 11012620 TI - Intrapulmonary concentrations of inflammatory cytokines in a mouse model of chronic respiratory infection caused by Pseudomonas aeruginosa. AB - We investigated the role of inflammatory cytokines in a mouse model of chronic Pseudomonas aeruginosa infection mimicking diffuse panbronchiolitis (DPB), and determined the effects of clarithromycin therapy on the production of these cytokines. The concentrations of IL-1beta, IL-2, IL-4, IL-5, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) were measured serially in the lungs of mice with experimentally induced chronic respiratory P. aeruginosa infection until 60 days after inoculation. The concentrations of these cytokines during the course of the disease were significantly higher than baseline (before inoculation, P<0.01 for all cytokines). Clarithromycin significantly inhibited the production of IL-1beta and TNF-alpha in the lung (P<0.01). The same treatment also reduced the levels of other cytokines, albeit insignificantly. Treatment with anti-TNF-alpha antibody significantly reduced the number of pulmonary lymphocytes and concentration of IL-1beta in the lung (P<0.01), but did not change the number of viable bacteria. Our findings resemble those detected in bronchoalveolar lavage fluid of patients with DPB and indicate that inflammatory cytokines play an important role in chronic P. aeruginosa lung infection. Our results also show that macrolides modulated the production of these cytokines, ultimately reducing lymphocyte accumulation in the lung. Our data suggest that anti-TNF-alpha antibody might be a useful new strategy for the treatment of chronic respiratory P. aeruginosa infection. PMID- 11012622 TI - Tuberculosis (TB) and HIV infection are independently associated with elevated serum concentrations of tumour necrosis factor receptor type 1 and beta2 microglobulin, respectively. AB - The aim of this study was to identify immune markers that are independently associated with HIV infection or TB in vivo. Using commercially available assays, we measured concentrations of five immune markers in sera from 175 out-patients attending medical clinics in Cote D'Ivoire and Ghana, West Africa. Patients were categorized into groups with TB only (TB+HIV-, n = 55), TB and HIV co-infection (TB+HIV+, n = 50), HIV infection only (TB-HIV+, n = 35), or neither infection (TB HIV-, n = 35). TB+HIV+ and TB-HIV+ groups were matched for blood CD4+ lymphocyte count. Mean +/- s.d. concentrations of beta2-microglobulin were similarly increased in both the TB-HIV+ (5.3+/-2.1 microg/ml, P<0.0001) and the TB+HIV+ (5.0+/-1.5 microg/ml, P<0.0001) groups compared with the TB-HIV- group (2.2+/-1.8 microg/ml), but were only slightly increased in the TB+HIV- group (3.2+/-1.8 microg/ml, P = 0.01). In contrast, mean serum concentrations of soluble tumour necrosis factor receptor type I (sTNF-RI) were similarly elevated in the TB+HIV- (1873+/-799 pg/ml, P<0.0001) and TB+HIV+ (1797+/-571 pg/ml, P<0.0001) groups compared with uninfected subjects (906+/-613 pg/ml), but there was only a small increase in sTNF-RI in the TB-HIV+ group (1231+/-165 pg/ml, P = 0.03). Both TB and HIV infection were associated with substantial elevation of serum concentrations of soluble CD8, soluble CD54, and sTNF-R type II. Analysis of additional samples from groups of TB+HIV- and TB+HIV+ patients receiving anti-TB treatment showed significant and equal reductions in mean serum sTNF-RI concentrations, but no significant change in mean beta2-microglobulin. Thus, serum beta2-microglobulin and sTNF-RI serve as relatively independent markers of HIV infection and TB, respectively, in studies of co-infected persons. PMID- 11012621 TI - A single measurement of CD38CD8 cells in HIV+, long-term surviving injecting drug users distinguishes those who will progress to AIDS from those who will remain stable. AB - This study compares the predictive power of a single measurement of CD8+CD38+, CD8+CD45RO+ or CD8+CD38+CD45RO+ subpopulations in predicting progression to AIDS in a cohort of HIV+ long-term surviving injecting drug users. The results showed that both the total CD8+ percentage, and the CD8+CD38+ and CD8+CD38+CD45RO+ subpopulations of cells all individually predicted progression to AIDS. In combination with CD4, only the CD8+CD38+ subpopulation enhanced the predictive power of the CD4 percentage alone. The CD8+ percentage correlated negatively with the CD4 percentage and the CD8+CD45RO+ subpopulation did not predict disease progression. The proportion of CD8+CD38+ cells identified which patients with a moderate CD4 level were more likely to progress to AIDS, and conversely, which patients with a low CD4 count were likely to remain clinically stable. The results were consistent irrespective of whether time was measured from the date of seroconversion, or from the date of the test. This study is the first to measure these markers in HIV-infected injecting drug users, and in long-term survivors. The results demonstrate the considerable added value of the CD8+CD38+ cell percentage over the CD4 count alone, in predicting HIV clinical progression. PMID- 11012623 TI - Efficient generation of respiratory syncytial virus (RSV)-neutralizing human MoAbs via human peripheral blood lymphocyte (hu-PBL)-SCID mice and scFv phage display libraries. AB - RSV is one of the major causes of pneumonia and bronchiolitis in infants and young children and is associated with high mortality. RSV neutralizing human antibody (hu-Ab) is known to mediate resistance to viral infection as well as to be an effective treatment for severe lower respiratory tract RSV infection. We have previously demonstrated that human primary and secondary immune responses can be established in severe combined immunodeficient mice engrafted with human peripheral blood lymphocytes (hu-PBL-SCID). By combining this animal model with the single-chain Fv antibody (scFv) phage display library technique, we were able to investigate further its clinical potential by generating a panel of human scFvs that exhibit both high F glycoprotein (RSV-F) binding affinities ( approximately 108 M(-1)) and strong neutralizing activities against RSV infection in vitro. Sequencing analysis of the randomly isolated anti-RSV-F scFv clones revealed that they were derived from different VH families with mutations in the complementarity-determining region 1 (CDR1). The results suggest that: (i) RSV-F specific human immune responses and affinity maturation can be induced in hu-PBL SCID mice; and (ii) this approach can be applied to generate large numbers of human scFvs with therapeutic potential. Despite the fact that hu-PBL-SCID mouse and human scFv phage display library have individually been established, our approach contributes a simple and significant step toward the generalization of antigen-specific human monoclonal antibody (hu-MoAb) production and their clinical applications. PMID- 11012624 TI - Influence of age and HLA type on interferon-gamma (IFN-gamma) responses to a naturally occurring polymorphic epitope of Plasmodium falciparum liver stage antigen-1 (LSA-1). AB - Antigenic polymorphism and HLA restriction may limit the immunogenicity of a subunit vaccine against liver-stage Plasmodium falciparum. We examined 59 clinical isolates and five laboratory clones of P. falciparum for polymorphism in the N- and C-terminal regions of LSA-1, evaluated binding of the corresponding peptides to selected HLA class I alleles, and measured IFN-gamma responses in residents of a malaria-endemic area of Papua New Guinea where HLA-A*1101, -24, B13, and -B40 are the most common class I alleles. LSA-1 polymorphism was limited to a single non-synonymous mutation encoding serine (S), proline (P), or threonine (T) at amino acid 85. Nine-mer 84-92 peptides with S, T, or P at the primary anchor position bound differentially to HLA-A11, -A2, and -B7. IFN-gamma ELISPOT responses increased with age in malaria-exposed subjects: 14-16% and 30 36% of 2-5- and 6-54-year-olds, respectively, had > or =10 IFN-gamma-secreting cells/106 peripheral blood mononuclear cells when stimulated with at least one peptide variant (P<0.05). IFN-gamma responses to all three peptides were also greater for older than younger individuals. No children < 3 years old had lymphocytes that responded to all three 84-92 peptides, whereas 45% of adults (mean age 48 years) had aggregated IFN-gamma responses. These data support the notion that age-related cumulative exposure to P. falciparum increases the frequency of IFN-gamma responses to polymorphic epitopes of liver-stage antigens such as LSA-1. PMID- 11012625 TI - Humoral hyporesponsiveness to a conjugate contraceptive vaccine and its bypass by diverse carriers using permissible adjuvant. AB - A contraceptive vaccine directed against human chorionic gonadotropin (hCG) has previously undergone clinical testing that demonstrated the feasibility of the approach in preventing pregnancy in women. Some immunized volunteers however, did not respond with an adequate anti-hCG antibody response despite employing highly immunogenic bacterial toxoids as carriers. Since there is some evidence that T cell responses to a complex protein typically focus on a few immunodominant epitopes, we investigated the responsiveness to hCG in mice of different haplotypes using the protein carrier diphtheria toxoid (DT). Our data showed a differential carrier effect of DT. With the aim of making a more potent immunogen employing promiscuous pathogen-derived Th peptides as carriers, peptide:antigen stoichiometric ratios were optimized. When tested individually using alum as the adjuvant, three such peptide conjugates improved the anti-hCG response, though not consistently to levels higher than the DT conjugate. Immunization with a combination of these synthetic epitopes generated anti-hCG responses higher than those achieved with DT or with the individual peptides. Antibodies were of high affinity and capable of neutralizing the bioactivity of hCG, but were devoid of anti-peptide reactivity. These results support our view that differential hyporesponsiveness in a diverse population may arise from inadequate carrier effect and that it can be overcome by use of pathogen-derived broadly reactive non-B Th epitopes employing only alum, a permissible adjuvant. PMID- 11012626 TI - Local bioactive tumour necrosis factor (TNF) in corneal allotransplantation. AB - The aim of this study was to examine the kinetic profile of bioactive TNF levels in aqueous humour of rabbit eyes undergoing corneal allograft rejection and to investigate the effect of locally blocking TNF activity after corneal transplantation. In a rabbit corneal transplantation, endothelial allograft rejection was identified and correlated with increase in central graft thickness. Samples of aqueous humour obtained on alternate days following transplantation were tested for TNF mRNA and bioactive TNF protein. To investigate the effect of locally blocking TNF activity in allograft recipients, the fusion protein TNFR-Ig was administered by injections into the anterior chamber after transplantation. Pulsatile increases in levels of this cytokine were found in 14 of 15 allograft recipients. Peaks of TNF bioactivity preceded by varying intervals the observed onset of rejection in allograft recipients. TNF levels were not elevated in aqueous humour from corneal autograft recipient controls or in serum of allografted animals. mRNA levels were elevated before onset of and during clinically observed allograft rejection. In three of seven animals receiving TNFR Ig injections on alternate days from day 8 to day 16 post-transplant, clear prolongation of corneal allograft survival was demonstrated. Bioactive TNF is present in aqueous humour following rabbit corneal allotransplantation. Rather than correlating directly with endothelial rejection onset, pulsatile peak levels of TNF precede and follow the observed onset of endothelial rejection. Blockade of TNF activity prolongs corneal allograft survival in some animals, indicating that this cytokine may be a suitable target in local therapy of corneal allograft rejection. PMID- 11012627 TI - Antigen-specific T cell responses in human peripheral blood leucocyte (hu-PBL) mouse chimera conditioned with radiation and an antibody directed against the mouse IL-2 receptor beta-chain. AB - A weakness of the hu-PBL-SCID model for the study of human immune functions is the appearance of anergy and the consequent loss of T cell function. We demonstrate here that human T cells retain normal functions during the early stage of chimerism. At 1 and 2 weeks post-engraftment, T cells isolated from the peritoneal cavity of hu-PBL chimeras could be activated and proliferated upon stimulation with phytohaemagglutinin (PHA) or specific antigens to which the cell donor was known to be immune. T cells derived from hu-PBL-SCID and hu-PBL NOD/LtSz-scid (NOD/SCID) mice not only proliferated but also produced interferon gamma (IFN-gamma) and IL-5 following in vitro stimulation with tetanus toxoid (TT) or hepatitis B surface antigen (HBsAg). These antigen-specific T cells could only be demonstrated when cognate antigen was administered together with or immediately following the PBL transfer. Without an early rechallenge with antigen in vivo, no TT- or HBsAg-specific T cell responses could be elicited, showing the vulnerability and antigen-dependence of the T cell response. Vigorous anti-TT or anti-HBs responses could be observed in all chimeras. Administration of antigen together with the PBL graft enhanced the humoral anti-TT response in SCID and NOD/SCID mice but had little effect on the anti-HBs antibody response in NOD/SCID mice. These data confirm the observation that the B cell compartment in hu-PBL SCID chimera is largely antigen-independent and extend this to SCID/NOD. PMID- 11012628 TI - A pivotal role of cell-bound but not soluble CD4 molecules in full development of lupus-like manifestations in MRL-Fas(lprcg)/Fas(lprcg) mice. AB - The role of CD4 molecules in the autoimmune and lymphoproliferative syndrome caused by murine Fas mutations was studied using the novel systemic lupus erythematosus (SLE) model, MRL-Fas(lpr(cg))/Fas(lprcg) (MRL-lpr(cg)) mice, in combination with the novel mutant CD4 gene producing soluble CD4 (sCD4) instead of membrane-bound CD4 (mCD4). For this purpose, various autoimmune manifestations were compared among MRL-lpr(cg) mice homozygous (CD4slprcg), heterozygous (CD4s/mlpr(cg)), and wild-type (CD4mlpr(cg)) for the CD4 mutation. The mortality, glomerulonephritis, proteinuria, and lymphadenopathy were significantly ameliorated in CD4slprcg compared with CD4mlpr(cg) and CD4s/mlpr(cg) mice, both being comparable in these clinical characteristics. In parallel with the clinical improvement, the serum levels of immunoglobulin, anti-DNA antibodies, anti nuclear antibodies and immune complexes, and the extent of glomerular immune deposition, were significantly lower in the former. The results indicate that mCD4 is important and can not be replaced by sCD4 in full development of SLE-like manifestations, and suggest that CD4+ T cells may aggravate the autoimmune disease by stimulating autoreactive B cells to produce autoantibodies through their helper activity in Fas mutant models. The sCD4 levels in the serum and spleen elevated with the increased accumulation of B220+CD4-CD8- (double-negative (DN)) T cells in CD4slpr(cg) mice. This, together with the significantly milder lymphadenopathy associated with lower DN T cell contents in CD4slpr(cg) than CD4mlpr(cg) mice, implies that some of abnormal DN T cells may be derived from cells of the CD4 lineage. PMID- 11012630 TI - Methods for the detection of blood parasites. PMID- 11012629 TI - Tumour necrosis factor-alpha (TNF-alpha) enhances lymphocyte migration into rheumatoid synovial tissue transplanted into severe combined immunodeficient (SCID) mice. AB - Adhesion mechanisms play a major role in the recruitment of peripheral blood lymphocytes (PBL) which characteristically infiltrate rheumatoid arthritis (RA) synovium and other chronically inflamed tissues. Through a sequential series of complex integrated adhesion and signalling events, 'multistep model of migration', specific subsets of PBL are recruited into inflamed tissues. In this process both leucocyte receptors and microvascular endothelial (MVE) counter receptors play a critical role. The MVE in particular, during an inflammatory state, is the target of various inflammatory mediators that cause the up regulation of several cell adhesion molecules (CAM). One of the most important factors known to be a powerful inducer of MVE CAM is TNF-alpha. Conversely, blocking TNF-alpha causes a down-modulation of CAM expression. To test directly the capacity of TNF-alpha to induce cell migration into RA synovium we adapted a model in which synovial grafts were implanted into SCID mice subcutaneously. Using this model we demonstrate that: (i) transplants remain viable and become vascularized and fed by mouse subdermal vessels; (ii) the mouse vasculature connects to the transplant vasculature which maintains the ability to express human CAM; (iii) intragraft injections of TNF-alpha up-regulate the expression of human CAM, following the down-regulation which occurred 4 weeks post transplantation; and (iv) the up-regulation of graft CAM is associated with increased human PBL migration into the transplants. This study provides direct evidence in vivo of the capacity of TNF-alpha to induce cell migration. In addition, it provides the experimental background for the optimal use of this model. PMID- 11012631 TI - Automated counting of white and red blood cells in the cerebrospinal fluid. AB - The objective of this study was to examine to what extent the automated method of the Bayer H*2 instrument could replace the visual counting of white and red blood cells in cerebrospinal fluid. The number of white blood cells as well as the percentage of mononuclear and polymorphonuclear cells were counted in the 'Baso channel' (Research screen 3) whereas the number of red cells were registered as the 'R-count' (Research screen 1). All automated cell counts were compared to visual estimates. The automated count yielded reliable results down to 5 x 106 white blood cells/l and 5 x 108 red blood cells/l. In some samples 'noise' was present in the Baso channel. A correct white blood cell count could then be obtained by counting the cells directly as dots on the screen. It was possible to differentiate between polymorphnuclear cells and mononuclear cells at all WBC concentrations. The automated counting of cerebrospinal fluid can be performed without changing thresholds or sample volumes of the instrument. Thus, in the routine practice it will be possible to alternate between automated counting of whole blood samples and cerebrospinal fluid samples. PMID- 11012632 TI - Two colour analysis of reticulated platelets. PMID- 11012633 TI - Angiocentric nasal T/natural killer cell lymphoma: a single centre study of prognostic factors in 108 patients. AB - Angiocentric T cell/natural killer (NK) nasal lymphoma remains a rare clinical presentation in North America and Europe but is more common in Asia and Latin America. We have reviewed 108 cases of angiocentric T/NK cell lymphoma of the nasal cavity with a view to establishing prognostic factors. Most patients were high or high intermediate clinical risk and had additional poor prognostic factors such as bulky disease, high levels of beta 2 microglobulin, advanced stage and multiple extranodal involvement. At 8 years, overall survival was 82%, 90% and 84% for low-intermediate, high-intermediate and high clinical, respectively. Disease free survival was very similar: 79%, 83% and 80%, respectively. Multivariate analysis did not identify any factor influencing overall survival and disease-free survival. There was no evidence that the international prognostic index (IPI) was applicable in these patients and it appears that angiocentric T/NK cell lymphoma is an independent prognostic factor itself. PMID- 11012634 TI - Severe iron deficiency anaemia and stroke. AB - Neurological complications of severe anaemia in childhood are rare. We report a case of severe iron deficiency in a child of 23 months, presenting as an acute hemiparesis. PMID- 11012635 TI - Efficacy of low dose intravenous immunoglobulins for post-splenectomy treatment of autoimmune haemolytic anaemia in a patient with hereditary spherocytosis. AB - Summary Idiopathic autoimmune haemolytic anaemia developed in a patient with hereditary spherocytosis. The behaviour of some osmotic fragility tests throughout the illness and the efficacy of intravenous immunoglobulins in controlling autoimmune haemolysis which recurred post splenectomy are discussed PMID- 11012636 TI - The cutaneous side-effects of hydroxyurea. AB - The cutaneous side-effects of long-term hydroxyurea therapy are not widely known and only rarely reported. We report on a patient who developed widespread skin changes, including the recently recognized hydroxyurea dermopathy, during long term treatment with hydroxyurea for polycythaemia rubra vera. The time course of the clinical changes suggests that they result from direct toxicity of hydroxyurea on the basal layer of the epidermis and mucosal surfaces. We aim to increase clinical awareness of this problem. PMID- 11012637 TI - Acute myeloid leukaemia with trilineage myelodysplasia complicated by masked diabetes insipidus. AB - We describe a rare case of acute myeloid leukaemia with trilineage myelodysplasia complicated by central diabetes insipidus. In the present case, diabetes insipidus was masked by corticosteroid deficiency due to hypopituitarism and clinical symptoms presented after administering methylprednisolone. Although the remission of leukaemia was not achieved by chemotherapy, excessive urinary output was well-controlled by nasal administration of 1-desamino-8-D-arginine vasopressin (DDAVP) during the course. PMID- 11012638 TI - Natural killer-cell lymphocytosis and strongyloides infection. AB - We report a case of strongyloides infection in a 72-year-old man presenting with acute angio-oedema and urticaria. He was also found to have natural killer cell (NK) large granular lymphocytosis (LGL). We discuss the possible relationship between the strongyloides infection and the NK-LGL lymphocytosis. PMID- 11012639 TI - Multiple myeloma and immune thrombocytopenia. AB - Immune thrombocytopenia (ITP) is frequently encountered in patients with lymphoproliferative disorders. However this is only rarely reported in patients with multiple myeloma. We describe three cases who presented initially with the clinical manifestations of ITP but were subsequently found to have multiple myeloma. Platelet count increments to standard treatment modalities for ITP were observed in all three patients with transient or partial response. The importance of recognizing the immune mediated thrombocytopenia in patients with myeloma and the implications of this combination are discussed. PMID- 11012640 TI - Laboratory findings associated with thrombophilia are not more common in inflammatory bowel disease. AB - Thromboembolic disease (TED) has been recognized as a complication of inflammatory bowel disease (IBD) since the 1930s (Bargen & Barker 1936). The relative contributions of inherited or acquired thrombophilia and the inflammatory response to the mechanism of this tendency is unclear. Thrombotic events are more common in active disease although significant numbers also occur spontaneously, when the disease is in clinical remission (Talbot et al. 1986; Jackson et al. 1997). Studies looking at the prevalence of specific thrombophilic states such as Antithrombin III deficiency (Jackson et al. 1997; Lake, Stauffer & Stuart 1978; Cianco et al. 1996; Ghosh et al. 1983), Factor V Leiden mutation (APC Resistance) (Jackson et al. 1997; Probert et al. 1997; Ardizzone et al. 1998; Liebman et al. 1998), anticardiolipin antibodies (Ciancio et al. 1996), Protein C (Wyshock, Caldwell & Crowley 1988; Korsten & Reis 1992) and Protein S deficiencies (Jorens et al. 1990; Aadland et al. 1992) in IBD have been contradictory or equivocal. We had previously found that IBD patients with a history of TED are not more likely to have a laboratory thrombophilic abnormality than those with uncomplicated disease. We also demonstrated that the prevalence of heterogenous laboratory thrombophilic abnormalities (usually minor) in all IBD patients may be as high as 60%, much higher than the recognized prevalence of TED (Lim, Jones & Gould 1996). We wondered how this would compare with the healthy non-IBD population. We have therefore explored the prevalence of such thrombophilic abnormalities in a group of IBD patients who had no history of TED and compared them with healthy age and sex matched controls. PMID- 11012641 TI - Thromboembolism in a patient with transient eosinophilia and thrombocytopenia. AB - A 24-year-old woman with an unremarkable medical history who developed bilateral deep venous thrombosis and pulmonary emboli is presented. Associated findings were severe eosinophilia and moderate thrombocytopenia. Since the major acquired and hereditary thrombogenic disorders were ruled out in this case (including antiphospholipid syndrome and heparin-induced thrombocytopenia), we believe that the severe eosinophilia per se could be the pro-coagulant factor leading to thrombosis and embolism in our patient. The role of eosinophilia in thrombosis is discussed. PMID- 11012642 TI - More effective ways of health care service delivery in the UK. PMID- 11012643 TI - The nose and nitric oxide: a review. PMID- 11012644 TI - Paediatric airway stenosis: laryngotracheal reconstruction or cricotracheal resection? AB - Modern surgical management of paediatric laryngotracheal stenosis includes a wide variety of surgical procedures. These can broadly be divided into two groups. First, laryngotracheal reconstruction (LTR) procedures in which the cricoid cartilage is split and the framework is expanded with various combinations of cartilage grafts and stents; and second, cricotracheal resection (CTR) where a segmental excision of the stenotic segment is done and an end-to-end anastomosis is performed. In this article we review the literature and our experience and discuss the relative indications for CTR and LTR in children. High decannulation rates have been reported for CTR; however, it remains a more extensive procedure than LTR involving extensive tracheal mobilization. If the tracheostomy site is included in the resection then a significant length of trachea is excised. Alternatively, LTR with cartilage grafting can precisely correct a specific stenosis with minimum morbidity and high decannulation rates for grade 2 and selected grade 3 stenosis. For the more severe stenosis treatment with LTR has been less successful. Retrospective data from this institution suggests that the children with grade 4 stenosis treated with LTR are more likely to require a subsequent open procedure to achieve decannulation than those treated with CTR. LTR is a less extensive procedure and is preferred for grade 2, selected grade 3 stenosis. CTR is the preferred option for grade 4 and severe grade 3 stenosis with a clear margin between the stenosis and the vocal cords. PMID- 11012645 TI - Sjogren's syndrome. PMID- 11012646 TI - Powered laryngeal shavers and laryngeal papillomatosis: a preliminary report. AB - Carbon dioxide (CO2) laser ablation is widely accepted as the most effective first line surgical treatment of recurrent laryngeal papilloma. However, vaporization of papilloma with the CO2 laser exposes the patient and staff to a potentially infective plume. Furthermore, unsuitable pathological tissue specimens and scarring of the surrounding laryngeal architecture are inevitable consequences of the thermal energy generated by this instrument. Following their innovative use in endonasal surgery, powered laryngeal microresecting instruments (Xomed Shavers) have been developed for laryngeal surgery. We assessed the value of these new instruments in five patients who had previously undergone laser ablation. All of the patients reported that their postoperative recovery was the 'same as' or 'better than' that following laser surgery. The surgeon reported that though there was more bleeding, shaver excision of papilloma provided good clearance of disease; easy collection of pathological samples; and a potentially safer and faster alternative to laser excision. PMID- 11012647 TI - The distribution of hypopnoeic events in relation to apnoeic events in patients with sleep-induced upper airway narrowing. AB - The purpose of this study was to investigate patients with sleep-induced upper airay narrowing to see if: (1) apnoeic and hypopnoeic events regularly coexist; (2) if sleep-induced upper airway narrowing usually involves more than one level; and (3) whether the distribution of hypopnoeic events reflects that of apnoeic events. A retrospective review of 50 patients with sleep-induced upper airway narrowing was conducted. Recordings were made with a solid-state microtransducer pressure-catheter and an external thermistor for assessment of the distribution and number of obstructive events. Ninety-four per cent of the patients had a mixture of apnoeic and hypopnoeic events. Both transpalatal and subpalatal obstructions were identified in 84% of patients. In 80% of the patients the level of apnoeic events, classified as either mainly (> 50% of total) transpalatal (upper) or subpalatal (lower) reflected that of hypopnoeic events. The percentage upper to total amount of hypopnoeic events correlated significantly with that of apnoeic events (R = 0.77, P < 0.001; Spearman rank correlation). PMID- 11012648 TI - Recurrence and malignant degeneration of 89 cases of inverted papilloma diagnosed in a non-tertiary referral population between 1975 and 1995: clinical predictors and p53 studies. AB - Eighty-nine patients with sinonasal inverted papilloma presenting between 1975 and 1995 were reviewed with the aims of studying predictors of tumour behaviour and correlating outcome with p53 expression. Correlation of clinical, radiological features and p53 status was made using chi2 and multiple logistic regression analysis with recurrence and malignant degeneration as the main outcome measures. Two patients had synchronous malignancy but no malignant degeneration was seen. There was no significant difference in recurrence between minor intranasal procedures and more extensive surgery for the first event. Younger patients were more likely to recur. (P = 0.0493, odds ratio 0.43). Those who smoked showed a trend towards multiple recurrence. p53 was expressed in 41% but did not predict recurrence. Morbidity was related to the extent of surgery. Inverted papilloma presenting to a non-tertiary centre is more benign than previously reported. Initial management by less extensive endoscopic surgery may reduce morbidity. PMID- 11012649 TI - Head and neck basal cell carcinoma: treatment using a 2-mm clinical excision margin. AB - Treatment of basal cell carcinoma (BCC) should completely remove the tumour whilst preserving the maximum amount of normal surrounding skin. Therefore, treatment is a compromise between safe excision margins and obtaining a satisfactory cosmetic result. We report the results of a prospective study of 63 patients who underwent excision of well-demarcated BCCs in the head and neck region. The surgical excision margin used was 2 mm. Histological assessment confirmed complete excision in 95% and there was no evidence of recurrence of the BCC over a 24-month follow-up period in all patients. We propose that a clinical excision margin of 2 mm is adequate for treatment of simple, well demarcated BCCs arising in the head and neck. PMID- 11012650 TI - Management of posterior epistaxis by endoscopic clipping of the sphenopalatine artery. AB - Posterior epistaxis poses a challenge to the otolaryngologist as the bleeding point itself cannot easily be identified. Haemostasis by conventional means is usually nasal packing and this results in repeated/persistent haemorrhage, morbidity and prolonged bed occupancy. In recent years the increased availability of rigid endoscopes and a better understanding of the anatomy of the nasal cavity have facilitated a direct approach to the sphenopalatine artery. Using a 0 degrees or 30 degrees rigid nasendoscope the sphenopalatine artery-the main blood supply to the nose-can be clearly identified and treated. Over the last 12 months we have employed endoscopic intranasal clipping of the said artery under a general anaesthetic to control persistent posterior nasal bleeding. Twelve patients have undergone 14 procedures within 48 h of failure of their conservative management. In all the epistaxis was controlled no complications were noted. The average follow-up period in our series was nine months. We believe that intranasal endoscopic clipping of the sphenopalatine artery is effective and less traumatic than either any other site of arterial ligation technique or repeated packing. In this series we employed only clipping of the artery and not diathermy/electrocautery to reduce the theoretical risk to adjacent structures PMID- 11012651 TI - Mandibular osteoradionecrosis: clinical behaviour and diagnostic aspects. AB - We have retrospectively studied 73 patients with mandibular osteoradionecrosis (ORN) following radiotherapy (high voltage external beam radiation alone, or combined with brachytherapy) ranging from 50 to 130 Gy (mean 76) with fields including the mandible. Based on this analysis we defined four distinctive stages: Stage 0: mucosal defects only; Stage I: radiological evidence of necrotic bone with intact mucosa; Stage II: positive radiographic findings with denuded bone intra-orally; Stage III: clinically exposed radionecrotic bone, verified by imaging techniques, along with skin fistulas and infection. The clinical features and characteristics of these stages are described, along with their clinical courses and outcome. The incompleteness of current ORN definitions, mainly emphasizing mucosal defects and bone exposure, and the need for a revised ORN classification is pointed out. Based on these observations and the insufficiency of previous definitions, we arrived at the following ORN definition: Radiological evidence of bone necrosis within the radiation field, where tumour recurrence has been excluded. PMID- 11012652 TI - Brain herniation and chronic otitis media: diagnosis and surgical management. AB - Herniation of the brain into the middle ear is a rare, but potentially life threatening complication of chronic otitis media. Fifty patients with a tegmen defect associated with chronic otitis media were operated on between 1985 and 1998. Among these 50, 15 patients presented brain herniation associated with the bony defect. Fourteen patients had undergone previous mastoid surgery for chronic otitis media. Neurological symptoms were encountered in five patients. In 10, magnetic resonance imaging (MRI) was performed before surgery, and a diagnosis of brain herniation could be made. The hernia was repaired in all patients using a middle fossa craniotomy, combined with a transmastoid approach in 11 cases where a large hernia, and/or inflammatory tissues were present in the mastoid. The herniated brain tissue was resected in all, and the dural and bony defects were closed with fascia and bone. No complication or recurrence occurred, during a mean follow-up of 2 years. In conclusion, the occurrence of severe neurological complications as a consequence of brain herniation emphasizes the necessity for recognition and appropriate management of this disease. Computerized tomography (CT) scanning allows the suspicion of brain herniation, but a definitive diagnosis can best be established with an MRI study. The hernia should be repaired using a middle fossa craniotomy, combined with a transmastoid approach in one or two stages, when necessary. PMID- 11012653 TI - Age-specific size of the normal adenoid pad on magnetic resonance imaging. AB - Conclusions regarding the significance and appearance of the adenoids incidentally noted on magnetic resonance (MR) scans of the brain largely rely on observations of previously published plain film data. In order to determine the age specific appearance of normal adenoid tissue as measured on sagittal T1 weighted midline MR images, we evaluated 189 patients without a history or clinical evidence of adenoid disease, who were sequentially referred for an MR scan of the brain. The thickness of the adenoid pad was measured to the nearest 1 mm along a line through the pharyngeal tubercle constructed perpendicular to the anterior clival surface. Patients were grouped according to age. Normal subjects demonstrated an age specific variation in the size of the pad with the maximal size being attained in early childhood and then steadily decreasing in later childhood and adulthood (P = 0.0001). The adenoids were largest in the 7-10 years age group (mean, 14.59 mm) and steadily declined to 4.83 mm by 60 years of age. Previous surgery had no effect on adenoid measurement (P = 0.582). Magnetic resonance scans provide an excellent method for assessing the adenoid pad. PMID- 11012654 TI - Performance data and survival in head and neck cancer. AB - A retrospective longitudinal investigation of 2701 patients extending from 1963 was conducted to study the effect of performance status on survival, locoregional recurrence and associations with host and tumour factors. It was found that those patients who are physically fit tended to have smaller tumours, less risk of neck node metastases and more chance of laryngeal cancer. In patients with advanced disease, locoregional recurrence did not appear to be more common in those with poor general condition but the death rate from the tumour appeared to increase as the general physical condition decreased. It is assumed this is due to the development of distant metastases. There is evidence that immunity is less well developed in those with poor general condition and this in turn, may be due to alcohol abuse and poor general nutrition. PMID- 11012655 TI - Evaluation of the Herbst Mandibular Advancement Splint in the management of patients with sleep-related breathing disorders. AB - Sleep-related breathing disorders such as snoring and obstructive sleep apnoea syndrome are the cause of significant social disruption and hypersomnolence. Several intraoral appliances for the treatment of these disorders have been described, especially where nasal continuous positive airway pressure is poorly tolerated. Mandibular Advancement Splints, such as the Herbst splint used in this study can also be offered to patients with mild to moderate sleep apnoea and simple snorers. The success and compliance rate noted in the literature are quite diverse. Few side-effects have been reported. We therefore undertook this study to assess: (1) the compliance; (2) the effectiveness; and (3) the side-effects in the long- and short-term. All patients for whom a Herbst splint had been prescribed in the last 18 months were sent a postal questionnaire regarding the above mentioned issues. In all, 179 questionnaires were posted and on analysis of the 132 returned it was noted that 82% of splints were worn and 88% of patients found the device to be effective. The long-term side-effects were minimal. In addition objective assessment on 10 patients with and without a jaw-retaining device was also obtained. We conclude that the Herbst Mandibular Advancement Splint is a justifiable option in selected subjects with sleep-related breathing disorders. PMID- 11012656 TI - A randomised controlled trial of the effect of regional nerve blocks on immediate post-tonsillectomy pain in adult patients. AB - Post-operative pain is the main cause of morbidity following tonsillectomy. The efficacy of glossopharyngeal and lesser palatine nerve blocks in controlling postoperative pain in adult patients was investigated prospectively. Patients 16 years and older admitted for elective tonsillectomy were randomised to one of three groups to receive a pre-incisional oropharyngeal injection of 0.5% bupivicaine, a 'dummy' injection of saline or no injection. Dissection tonsillectomy and general anaesthetic techniques were standardized. Postoperative pain was monitored for 24 h. ANOVA, chi2 and Fisher's exact test were used for intergroup comparisons. Ninety-two patients (72 women and 20 men), mean age 22 years were studied. Twenty-nine patients received 0.5% bupivicaine, 30 saline and 33 no pre-incisional injection. The overall mean pain scores of 2.1, 1.9 and 1.9 in the bupivicaine, saline and no injection groups were similar. Glossopharyngeal and lesser palatine 0.5% bupivicaine nerve blocks are not effective in reducing early post-tonsillectomy pain. PMID- 11012657 TI - The effect of Isshiki type 1 thyroplasty on quality of life and vocal performance. AB - Type 1 thyroplasty for unilateral vocal cord palsy improves many vocal outcome measures but there is little information on quality of life despite its increasingly recognized importance. Our prospective study examined its effect on a range of subjective and objective measures and quality of life. Twenty-seven patients underwent thyroplasty. Before and after surgery they completed a vocal performance questionnaire and the Nottingham Health Profile (NHP); instrumental analyses of jitter, shimmer and noise-harmonic ratio (NHR); and perceptual analyses of grade, roughness, breathiness, aesthenia and strain (GRBAS) were also performed. Significant improvements were found in instrumental, perceptual and self-assessment of voice and the energy, social and emotional dimensions of the NHP. Three patients had initially poor results but were successfully revised. These results of type 1 thyroplasty compare favourably with those previously published. The improvement in quality of life appears to result directly from improved voice. Many thyroplasty patients have limited life expectancy: early surgical intervention should be considered. PMID- 11012658 TI - Acoustic rhinometric evaluation of the nasal response to exercise in patients with nasal septal deviation. AB - We aimed to investigate nasal response to exercise in patients with a nasal septal deviation. Acoustic rhinometric measurements of cross-sectional areas (CSA) at 3.3, 4.0, 6.4 cm from the nostril, and nasal volume (NV) were conducted in 10 patients. The consecutive measurements were undertaken just before and immediately after 10, 20, and 30 min of treadmill exercise. In the concave nasal cavities, NV and CSA at 3.3 cm and 6.4 cm from the nostril showed a significant increase in immediate post-exercise and 10-minute post-exercise measurements. At 4.0 cm from the nostril, a significant increase was sustained by 20 min after exercise. However, in convex nasal cavities, exercise did not result in a significant increase except for the immediate post-exercise measurement at 4 cm from the nostril. The results of this study indicated that, in nasal septal deviation, the mucosal response is more prominent in the concave nasal cavities than in the convex nasal cavities. PMID- 11012659 TI - Spontaneous resolution of tonsillitis in children on the waiting list for tonsillectomy. AB - The aim of this study was to determine whether children undergo spontaneous resolution of recurrent acute tonsillitis whilst awaiting surgery. Eighty children who had been on the waiting list for tonsillectomy for at least 9 months were requested to attend a review clinic with a second review 6 months later if appropriate. Nineteen (27%) of the 70 children who completed the study no longer warranted surgery and were removed from the waiting list. We conclude that those children who have been on the waiting list for a long period of time should be reassessed prior to the planned tonsillectomy to see if surgery is still indicated. PMID- 11012660 TI - Thioredoxin-thioredoxin reductase--a system that has come of age. PMID- 11012661 TI - Physiological functions of thioredoxin and thioredoxin reductase. AB - Thioredoxin, thioredoxin reductase and NADPH, the thioredoxin system, is ubiquitous from Archea to man. Thioredoxins, with a dithiol/disulfide active site (CGPC) are the major cellular protein disulfide reductases; they therefore also serve as electron donors for enzymes such as ribonucleotide reductases, thioredoxin peroxidases (peroxiredoxins) and methionine sulfoxide reductases. Glutaredoxins catalyze glutathione-disulfide oxidoreductions overlapping the functions of thioredoxins and using electrons from NADPH via glutathione reductase. Thioredoxin isoforms are present in most organisms and mitochondria have a separate thioredoxin system. Plants have chloroplast thioredoxins, which via ferredoxin-thioredoxin reductase regulates photosynthetic enzymes by light. Thioredoxins are critical for redox regulation of protein function and signaling via thiol redox control. A growing number of transcription factors including NF kappaB or the Ref-1-dependent AP1 require thioredoxin reduction for DNA binding. The cytosolic mammalian thioredoxin, lack of which is embryonically lethal, has numerous functions in defense against oxidative stress, control of growth and apoptosis, but is also secreted and has co-cytokine and chemokine activities. Thioredoxin reductase is a specific dimeric 70-kDa flavoprotein in bacteria, fungi and plants with a redox active site disulfide/dithiol. In contrast, thioredoxin reductases of higher eukaryotes are larger (112-130 kDa), selenium dependent dimeric flavoproteins with a broad substrate specificity that also reduce nondisulfide substrates such as hydroperoxides, vitamin C or selenite. All mammalian thioredoxin reductase isozymes are homologous to glutathione reductase and contain a conserved C-terminal elongation with a cysteine-selenocysteine sequence forming a redox-active selenenylsulfide/selenolthiol active site and are inhibited by goldthioglucose (aurothioglucose) and other clinically used drugs. PMID- 11012662 TI - Thioredoxin reductase two modes of catalysis have evolved. AB - Thioredoxin reductase (EC 1.6.4.5) is a widely distributed flavoprotein that catalyzes the NADPH-dependent reduction of thioredoxin. Thioredoxin plays several key roles in maintaining the redox environment of the cell. Like all members of the enzyme family that includes lipoamide dehydrogenase, glutathione reductase and mercuric reductase, thioredoxin reductase contains a redox active disulfide adjacent to the flavin ring. Evolution has produced two forms of thioredoxin reductase, a protein in prokaryotes, archaea and lower eukaryotes having a Mr of 35 000, and a protein in higher eukaryotes having a Mr of 55 000. Reducing equivalents are transferred from the apolar flavin binding site to the protein substrate by distinct mechanisms in the two forms of thioredoxin reductase. In the low Mr enzyme, interconversion between two conformations occurs twice in each catalytic cycle. After reduction of the disulfide by the flavin, the pyridine nucleotide domain must rotate with respect to the flavin domain in order to expose the nascent dithiol for reaction with thioredoxin; this motion repositions the pyridine ring adjacent to the flavin ring. In the high Mr enzyme, a third redox active group shuttles the reducing equivalent from the apolar active site to the protein surface. This group is a second redox active disulfide in thioredoxin reductase from Plasmodium falciparum and a selenenylsulfide in the mammalian enzyme. P. falciparum is the major causative agent of malaria and it is hoped that the chemical difference between the two high Mr forms may be exploited for drug design. PMID- 11012663 TI - Thioredoxin reductase as a pathophysiological factor and drug target. AB - Human cytosolic thioredoxin reductase (TrxR), a homodimeric protein containing 1 selenocysteine and 1 FAD per subunit of 55 kDa, catalyses the NADPH-dependent reduction of thioredoxin disulfide and of numerous other oxidized cell constituents. As a general reducing enzyme with little substrate specificity, it also contributes to redox homeostasis and is involved in prevention, intervention and repair of damage caused by H2O2-based oxidative stress. Being a selenite reducing enzyme as well as a selenol-containing enzyme, human TrxR plays a central role in selenium (patho)physiology. Both dietary selenium deficiency and selenium oversupplementation, a lifestyle phenomenon of our time, appear to interfere with the activity of TrxR. Selenocysteine 496 of human TrxR is a major target of the anti-rheumatic gold-containing drug auranofin, the formal Ki for the stoichiometric inhibition being 4 nM. The hypothesis that TrxR and extracellular thioredoxin play a pathophysiologic role in chronic diseases such as rheumatoid arthritis, Sjogren's syndrom, AIDS, and certain malignancies, is substantiated by biochemical, virological, and clinical evidence. Reduced thioredoxin acts as an autocrine growth factor in various tumour diseases, as a chemoattractant, and it synergises with interleukins 1 and 2. The effects of anti tumour drugs such as carmustine and cisplatin can be explained in part by the inhibition of TrxR. Consistently, high levels of the enzyme can support drug resistance. TrxRs from different organisms such as Escherichia coli, Mycobacterium leprae, Plasmodium falciparum, Drosophila melanogaster, and man show a surprising diversity in their chemical mechanism of thioredoxin reduction. This is the basis for attempts to develop specific TrxR inhibitors as drugs against bacterial infections like leprosy and parasitic diseases like amebiasis and malaria. PMID- 11012664 TI - AhpF and other NADH:peroxiredoxin oxidoreductases, homologues of low Mr thioredoxin reductase. AB - A group of bacterial flavoproteins related to thioredoxin reductase contain an additional approximately 200-amino-acid domain including a redox-active disulfide center at their N-termini. These flavoproteins, designated NADH:peroxiredoxin oxidoreductases, catalyze the pyridine-nucleotide-dependent reduction of cysteine based peroxidases (e.g. Salmonella typhimurium AhpC, a member of the peroxiredoxin family) which in turn reduce H2O2 or organic hydroperoxides. These enzymes catalyze rapid electron transfer (kcat > 165 s-1) through one tightly bound FAD and two redox-active disulfide centers, with the N-terminal-most disulfide center acting as a redox mediator between the thioredoxin-reductase like part of these proteins and the peroxiredoxin substrates. A chimeric protein with the first 207 amino acids of S. typhimurium AhpF attached to the N-terminus of Escherichia coli thioredoxin reductase exhibits very high NADPH:peroxiredoxin oxidoreductase and thioredoxin reductase activities. Catalytic turnover by NADH:peroxiredoxin oxidoreductases may involve major domain rotations, analogous to those proposed for bacterial thioredoxin reductase, and cycling of these enzymes between two electron-reduced (EH2) and four electron-reduced (EH4) redox states. PMID- 11012665 TI - Catalytic differences between porcine blastocyst and placental aromatase isozymes. AB - Two isozymes of porcine aromatase, the placental and the blastocyst forms, were expressed in CHO cells using the mammalian cell transfection method. Using an 'in cell' assay (a 3H-water release method), catalytic parameters of the porcine placental aromatase were found to be very similar to those of the human enzyme; however, the activity of the blastocyst isozyme was found to be one-thirtieth that of the placental isozyme. Product isolation assay (using testosterone as the substrate) revealed that the major steroid products were 17beta-estradiol and 19 nortestosterone. The product ratio of estradiol/19-nortestosterone was found to be 94 : 6 for the porcine placental form, 6 : 94 for the porcine blastocyst form, and 92 : 8 for the human wild-type aromatase. Therefore, the porcine blastocyst aromatase isozyme catalyzes mainly androgen 19-desmethylation rather than aromatization. In addition, inhibition profile analyses on the placental and blastocyst isozymes were performed using three steroidal inhibitors [4 hydroxyandro-stenedione (4-OHA), 7alpha-(4'-amino)phenylthio-1, 4-androstandiene 3,17-dione (7alpha-APTADD), and bridge (2, 19-methyleneoxy) androstene-3,17-dione (MDL 101,003)], and four nonsteroidal inhibitors [aminoglutethimide (AG), CGS 20267, ICI D1033, and vorozole (R83842)]. While the two isozymes of porcine aromatase share 93% amino-acid sequence identity, our results indicate that the two porcine aromatase isozymes have distinct responses to various aromatase inhibitors. PMID- 11012666 TI - Cloning, expression and chromosomal localization of a novel human dipeptidyl peptidase (DPP) IV homolog, DPP8. AB - Dipeptidyl peptidase (DPP) IV has roles in T-cell costimulation, chemokine biology, type-II diabetes and tumor biology. Fibroblast activation protein (FAP) has been implicated in tumor growth and cirrhosis. Here we describe DPP8, a novel human postproline dipeptidyl aminopeptidase that is homologous to DPPIV and FAP. Northern-blot hybridization showed that the tissue expression of DPP8 mRNA is ubiquitous, similar to that of DPPIV. The DPP8 gene was localized to chromosome 15q22, distinct from a closely related gene at 19p13.3 which we named DPP9. The full-length DPP8 cDNA codes for an 882-amino-acid protein that has about 27% identity and 51% similarity to DPPIV and FAP, but no transmembrane domain and no N-linked or O-linked glycosylation. Western blots and confocal microscopy of transfected COS-7 cells showed DPP8 to be a 100-kDa monomeric protein expressed in the cytoplasm. Purified recombinant DPP8 hydrolyzed the DPPIV substrates Ala Pro, Arg-Pro and Gly-Pro. Thus recombinant DPP8 shares a postproline dipeptidyl aminopeptidase activity with DPPIV and FAP. DPP8 enzyme activity had a neutral pH optimum consistent with it being nonlysosomal. The similarities between DPP8 and DPPIV in tissue expression pattern and substrates suggests a potential role for DPP8 in T-cell activation and immune function. PMID- 11012667 TI - Functional regions in the essential light chain of smooth muscle myosin as revealed by the mutagenesis approach. AB - The endogenous essential light chain (LC17) of myosin from intestine smooth muscle was replaced with mutated essential light chains prepared using recombinant techniques. Complete exchange was observed with histidine-tagged derivatives of LC17a, LC17b and E122A-LC17a (LC17a and LC17b are the usual constituants of smooth muscle myosin), with small changes in the ATPase activity of reconstituted myosins. Much less exchange was observed with the light-chain derivative lacking the last 12 amino acid residues, demonstrating the importance of this segment, which may act as one arm of a pair of pincers to bind the myosin heavy chain. PMID- 11012668 TI - Dual metabolic pathway of 25-hydroxyvitamin D3 catalyzed by human CYP24. AB - Human 25-hydroxyvitamin D3 (25(OH)D3) 24-hydroxylase (CYP24) cDNA was expressed in Escherichia coli, and its enzymatic and spectral properties were revealed. The reconstituted system containing the membrane fraction prepared from recombinant E. coli cells, adrenodoxin and adrenodoxin reductase was examined for the metabolism of 25(OH)D3, 1alpha,25(OH)2D3 and their related compounds. Human CYP24 demonstrated a remarkable metabolism consisting of both C-23 and C-24 hydroxylation pathways towards both 25(OH)D3 and 1alpha,25(OH)2D3, whereas rat CYP24 showed almost no C-23 hydroxylation pathway [Sakaki, T. Sawada, N. Nonaka, Y. Ohyama, Y. & Inouye, K. (1999) Eur. J. Biochem. 262, 43-48]. HPLC analysis and mass spectrometric analysis revealed that human CYP24 catalyzed all the steps of the C-23 hydroxylation pathway from 25(OH)D3 via 23S, 25(OH)2D3, 23S,25,26(OH)3D3 and 25(OH)D3-26,23-lactol to 25(OH)D3-26, 23-lactone in addition to the C-24 hydroxylation pathway from 25(OH)D3 via 24R,25(OH)2D3, 24-oxo-25(OH)D3, 24-oxo 23S,25(OH)2D3 to 24,25,26,27-tetranor-23(OH)D3. On 1alpha,25(OH)2D3 metabolism, similar results were observed. These results strongly suggest that the single enzyme human CYP24 is greatly responsible for the metabolism of both 25(OH)D3 and 1alpha,25(OH)2D3. We also succeeded in the coexpression of CYP24, adrenodoxin and NADPH-adrenodoxin reductase in E. coli. Addition of 25(OH)D3 to the recombinant E. coli cell culture yielded most of the metabolites in both the C-23 and C-24 hydroxylation pathways. Thus, the E. coli expression system for human CYP24 appears quite useful in predicting the metabolism of vitamin D analogs used as drugs. PMID- 11012669 TI - Dual nature of the distal histidine residue in the autoxidation reaction of myoglobin and hemoglobin comparison of the H64 mutants. AB - The oxygenated form of myoglobin or hemoglobin is oxidized easily to the ferric met-form with generation of the superoxide anion. To make clear the possible role(s) of the distal histidine (H64) residue in the reaction, we have carried out detailed pH-dependence studies of the autoxidation rate, using some typical H64 mutants of sperm whale myoglobin, over the wide range of pH 5-12 in 0.1 M buffer at 25 degrees C. Each mutation caused a dramatic increase in the autoxidation rate with the trend H64V >/= H64G >/= H64L >> H64Q > H64 (wild-type) at pH 7.0, whereas each mutant protein showed a characteristic pH-profile which is essentially different from that of the wild-type or native sperm whale MbO2. In particular, all the mutants have lost the acid-catalyzed process that can play a dominant role in the autoxidation reaction of most mammalian myoglobins or hemoglobins. Kinetic analyses of various types of pH-profiles lead us to conclude that the distal histidine residue can play a dual role in the nucleophilic displacement of O2- from MbO2 or HbO2 in protic, aqueous solution. One is in a proton-relay mechanism via its imidazole ring, and the other is in the maximum protection of the FeO2 center against a water molecule or an hydroxyl ion that can enter the heme pocket from the surrounding solvent. PMID- 11012670 TI - Purification and primary structure of a new bovine spermadhesin. AB - Z13 is a new seminal plasma protein made up of two disulfide-linked 13-kDa subunits that was identified in our laboratory by 2D PAGE. In this report we present the purification of Z13 from bovine seminal plasma. In solution, the protein is a nonglycosylated dimer that presents one interchain disulfide bond and does not show heparin-binding properties. The complete primary structure and the localization of the S-S bridges are reported. The results suggest that Z13 is a new protein of the spermadhesin family whose members are thought to play a prominent role in different aspects of fertilization. PMID- 11012671 TI - Regulation of Gadd45gamma expression by C/EBP. AB - The Gadd45gamma (growth arrest and DNA damage-inducible) gene is activated transcriptionally by at least two kinds of agents: DNA damaging agent such as methyl methanesulfonate (MMS) and UV radiation, or cytokines such as interleukin (IL)-6, IL-2 and granulocyte colony-stimulating factor (G-CSF). To investigate the sequences and transcription factors involved in induction of Gadd45gamma after treatment with IL-6, the human gene was cloned and sequenced. We found C/EBP (CCAAT/enhancer-binding protein) family proteins, major transcription factors in the IL-6 signal transduction pathway, could regulate the transcriptional activity of the Gadd45gamma promoter. In addition, a noncanonical C/EBP-binding site within the Gadd45gamma promoter where C/EBPbeta and C/EBPdelta could bind, was identified by electrophoretic mobility shift assay (EMSA) and reporter gene analysis. Furthermore, we found a coordinated expression profile between Gadd45gamma mRNA and C/EBPs (beta and delta) protein during the differentiation of M1 cells: the amount of Gadd45gamma transcripts became maximal when both C/EBPbeta and C/EBPdelta levels were high, on day 1 of differentiation of M1 cells after treatment with IL-6. These findings suggest that mitotic growth arrest coupled to M1 cell differentiation is mediated by C/EBPs stimulation of growth arrest-associated genes such as Gadd45gamma. PMID- 11012672 TI - A masquerade-like serine proteinase homologue is necessary for phenoloxidase activity in the coleopteran insect, Holotrichia diomphalia larvae. AB - Previously, we reported the molecular cloning of cDNA for the prophenoloxidase activating factor-I (PPAF-I) that encoded a member of the serine proteinase group with a disulfide-knotted motif at the N-terminus and a trypsin-like catalytic domain at the C-terminus [Lee, S.Y., Cho, M.Y., Hyun, J.H., Lee, K.M., Homma, K.I., Natori, S. , Kawabata, S.I., Iwanaga, S. & Lee, B.L. (1998) Eur. J. Biochem. 257, 615-621]. PPAF-I is directly involved in the activation of pro phenoloxidase (pro-PO) by limited proteolysis and the overall structure is highly similar to that of Drosophila easter serine protease, an essential serine protease zymogen for pattern formation in normal embryonic development. Here, we report purification and molecular cloning of cDNA for another 45-kDa novel PPAF from the hemocyte lysate of Holotrichia diomphalia larvae. The gene encodes a serine proteinase homologue consisting of 415 amino-acid residues with a molecular mass of 45 256 Da. The overall structure of the 45-kDa protein is similar to that of masquerade, a serine proteinase homologue expressed during embryogenesis, larval, and pupal development in Drosophila melanogaster. The 45 kDa protein contained a trypsin-like serine proteinase domain at the C-terminus, except for the substitution of Ser of the active site triad to Gly and had a disulfide-knotted domain at the N-terminus. A highly similar 45-kDa serine proteinase homologue was also cloned from the larval cDNA library of another coleopteran, Tenebrio molitor. By in vitro reconstitution experiments, we found that the purified 45-kDa serine proteinase homologue, the purified active PPAF-I and the purified pro-PO were necessary for expressing phenoloxidase activity in the Holotrichia pro-PO system. However, incubation of pro-PO with either PPAF-I or 45-kDa protein, no phenoloxidase activity was observed. Interestingly, when the 45-kDa protein was incubated with PPAF-I and pro-PO in the absence, but not in the presence of Ca2+, the 45-kDa protein was cleaved to a 35-kDa protein. RNA blot hybridization revealed that expression of the 45-kDa protein was increased in the Holotrichia hemolymph after Escherichia coli challenge. PMID- 11012673 TI - Shifting the NAD/NADP preference in class 3 aldehyde dehydrogenase. AB - Among pyridine-nucleotide-dependent oxidoreductases, the class 3 family of aldehyde dehydrogenases (ALDHs) is unusual in its ability to function with either NAD or NADP. This is all the more surprising because an acidic residue, Glu140, coordinates the adenine ribose 2' hydroxyl. In many NAD-dependent dehydrogenases a similarly placed carboxylate is thought to be responsible for exclusion of NADP. The corresponding residue in most (approximately 71%) sequences in the ALDH extended family is also Glu, and most of these are NAD-specific enzymes. Site directed mutagenesis was performed on this residue in rat class 3 ALDH. Our results indicate that this residue contributes to tighter binding of NAD in the native enzyme, but suggest that additional factors must contribute to the ability to utilize NADP. Mutagenesis of an adjacent basic residue (Lys137) indicates that it is even more essential for binding both coenzymes, consistent with its conservation in nearly all ALDHs (> 98%). PMID- 11012674 TI - Spectroscopic probing of the influence of calcium and the gla domain on the interaction between the first EGF domain in factor VIIa and tissue factor. AB - The binding of factor VIIa (FVIIa) to tissue factor (TF) initiates blood coagulation. The binary complex is dependent on Ca2+ binding to several sites in FVIIa and is maintained by multiple contacts distributed throughout the various domains. Although the contributions from various residues and domains, including the Ca2+ coordination, to the global binding energy have been characterized, their importance for specific local interactions is virtually unknown. To address this aspect, we have attached four spectroscopic probes to an engineered Cys residue replacing Phe140 in soluble TF (sTF). This allows the monitoring of local changes in hydrophobicity and rigidity upon complex formation at the interface between the first epidermal growth factor-like (EGF1) domain of FVIIa and sTF. The fluorescent labels used sense a more hydrophobic environment and the spin labels are dramatically immobilized when FVIIa binds sTF. The results obtained with a 4-carboxyglutamic acid (Gla)-domainless derivative of FVIIa indicate that the Gla domain has no or minimal influence on the interaction between EGF1 and sTF. However, there is a difference in local Ca2+ dependence between Gla domainless and full-length FVIIa. PMID- 11012675 TI - Identification of novel prostate-specific antigen-binding peptides modulating its enzyme activity. AB - Prostate-specific antigen (PSA) is a serine protease with highly prostate specific expression. Measurement of PSA in serum is widely used for diagnosis and monitoring of prostate cancer. PSA dissolves the seminal gel forming after ejaculation. It has been suggested to mediate invasion and metastasis of prostate cancer but also to exert antiangiogenic activity. We have identified peptides specific for PSA by screening cyclic phage display peptide libraries. PSA-binding peptides were isolated from four different libraries and produced as a fusion protein with glutathione S-transferase (GST). The phage and fusion proteins were shown to bind to PSA specifically as indicated by lack of binding to other serine proteinases. A peptide with four cysteines showed the highest affinity for PSA. Zn2+, an inhibitor of PSA activity, increased the affinity of the peptides to PSA. The binding specificity was characterized by cross-inhibition using monoclonal anti-PSA antibodies of known epitope specificities. The peptides bound to the same region as mAbs specific for free PSA indicating that they bind close to the active site of the enzyme. The peptides enhanced the enzyme activity of PSA against a chromogenic substrate. These results show that peptides binding to PSA and modulating its enzyme activity can be developed by phage display technique. The peptides have the potential to be used for identification of PSA variants and for imaging and targeting of prostatic tumors. PMID- 11012676 TI - Kinetic evidences for facilitation of peptide channelling by the proteasome activator PA28. AB - The activation kinetics of constitutive and IFNgamma-stimulated 20S proteasomes obtained with homomeric (recPA28alpha, recPA28beta) and heteromeric (recPA28alphabeta) forms of recombinant 11S regulator PA28 was analysed by means of kinetic modelling. The activation curves obtained with increasing concentrations of the individual PA28 subunits (RecP28alpha/RecP28beta/RecP28alpha + RecP28beta) exhibit biphasic characteristics which can be attributed to a low-level activation by PA28 monomers and full proteasome activation by assembled activator complexes. The dissociation constants do not reveal significant differences between the constitutive and the immunoproteasome. Intriguingly, the affinity of the proteasome towards the recPA28alphabeta complex is about two orders of magnitude higher than towards the homomeric PA28alpha and PA28beta complexes. Striking similarities can been revealed in the way how PA28 mediates the kinetics of latent proteasomes with respect to three different fluorogenic peptides probing the chymotrypsin-like, trypsin-like and peptidylglutamyl-peptide hydrolyzing like activity: (a) positive cooperativity disappears as indicated by a lack of sigmoid initial parts of the kinetic curves, (b) substrate affinity is increased, whereby (c), the maximal activity remains virtually constant. As these kinetic features are independent of the peptide substrates, we conclude that PA28 exerts its activating influence on the proteasome by enhancing the uptake (and release) of shorter peptides. PMID- 11012677 TI - The assembly factor P17 from bacteriophage PRD1 interacts with positively charged lipid membranes. AB - The interactions of the assembly factor P17 of bacteriophage PRD1 with liposomes were investigated by static light scattering, fluorescence spectroscopy, and differential scanning calorimetry. Our data show that P17 binds to positively charged large unilamellar vesicles composed of the zwitterionic 1-palmitoyl-2 oleoyl-phosphatidylcholine and sphingosine, whereas only a weak interaction is evident for 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles. P17 does not bind to negatively charged membranes composed of 1-palmitoyl-2-oleoyl phosphatidylglycerol and 1-palmitoyl-2-oleoyl-phosphatidylcholine. Our differential scanning calorimetry results reveal that P17 slightly perturbs the phase behaviour of neutral phosphatidylcholine and negatively charged multilamellar vesicles. In contrast, the phase transition temperature of positively charged dimyristoylphosphatidylcholine/sphingosine multilamellar vesicles (molar ratio 9 : 1, respectively) is increased by approximately 2.4 degrees C and the half width of the enthalpy peak broadened from 1.9 to 5.6 degrees C in the presence of P17 (protein : lipid molar ratio 1 : 47). Moreover, the enthalpy peak is asymmetrical, suggesting that lipid phase separation is induced by P17. Based on the far-UV CD spectra, the alpha-helicity of P17 increases upon binding to positively charged micelles composed of Triton X-100 and sphingosine. We propose that P17 can interact with positively charged lipid membranes and that this binding induces a structural change on P17 to a more tightly packed and ordered structure. PMID- 11012678 TI - Interaction of the targeting sequence of chloroplast precursors with Hsp70 molecular chaperones. AB - We have analyzed the interaction of DnaK and plant Hsp70 proteins with the wild type ferredoxin-NADP+ reductase precursor (preFNR) and mutants containing amino acid replacements in the targeting sequence. Using an algorithm already developed [Rudiger, S., Germeroth, L., Schneider-Mergener, J. & Bukau, B. (1997) EMBO J. 16, 1501-1507] we observed that 75% of the 727 plastid precursor proteins analyzed contained at least one site with high likelihood of DnaK binding in their transit peptides. Statistical analysis showed a decrease of DnaK binding site frequency within the first 15 amino-acid residues of the transit peptides. Using fusion proteins we detected the interaction of DnaK with the transit peptide of the folded preFNR but not with the mature region of the protein. Discharge of DnaK from the presequence was favored by addition of MgATP. When a putative DnaK binding site was artificially added at the N-terminus of the mature protein, we observed formation of complexes with bacterial and plant Hsp70 molecular chaperones. Reducing the likelihood of DnaK binding by directed mutagenesis of the presequence increased the release of bound DnaK. The Hsp70 proteins from plastids and plant cell cytosol also interacted with the preFNR transit peptide. Overall results are discussed in the context of the proposed models to explain the organelle protein import. PMID- 11012679 TI - The rat kidney acylase I, characterization and molecular cloning. Differences with other acylases I. AB - The soluble acylase I from rat kidney was purified to homogeneity using a five step procedure. As the resulting protein was found to have a relative molecular mass of 125 kDa based on size-exclusion chromatography and 44 kDa based on SDS/PAGE, the native protein was taken to consist of three subunits. The amino acid sequence of a peptide resulting from limited proteolysis of the polypeptide chain with proteinase K, which was determined by microsequencing (RHEFHALRAGFALDEGLA), was found to be very similar to the corresponding sequence of porcine kidney acylase I. However, as N-furyl-acryloyl-L-methionine, a synthetic substrate for porcine acylases, was not hydrolyzed by the rat enzyme, it was suggested that the polypeptide chain might differ in other respects from those of the other acylases I. A full length cDNA coding for the rat kidney acylase I was therefore isolated and found to contain a 1224-bp open reading frame encoding a protein consisting of 408 amino-acid residues, which corresponded to a calculated molecular mass of 45 847 Da per subunit. The deduced amino-acid sequence showed 93.6% and 87.2% identity with that of the human liver and porcine kidney, respectively. PMID- 11012680 TI - Signal transduction pathways triggered by fibroblast growth factor receptor 1 expressed in Xenopus laevis oocytes after fibroblast growth factor 1 addition. Role of Grb2, phosphatidylinositol 3-kinase, Src tyrosine kinase, and phospholipase Cgamma. AB - Xenopus oocytes expressing fibroblast growth factor receptor 1 (FGFR1) were used as a biological model system to analyse the signal transduction pathways that are triggered by fibroblast growth factor 1 (FGF1). Germinal vesicle breakdown (GVBD) and phosphorylation of extracellular signal-regulated protein kinase 2 (ERK2) occured 15 h after FGF1 addition. These events were Ras-dependent as they were blocked by a Ras dominant negative form. The Ras activity was promoted by three upstream effectors, growth factor-bound protein 2 (Grb2), phosphatidylinositol 3 kinase (PI3K) and Src cytoplasmic kinase. Ras activation was inhibited by a Grb2 dominant negative form (P49L), by PI3K inhibitors, including wortmannin, LY294002, the N-SH2 domain of p85alpha PI3K and by the SH2 domain of Src. Src activation induced by FGF1 was blocked by the SH2 domain of Src and PP2, a specific inhibitor of Src. The Grb2 adaptor was recruited by the upstream Src homology 2/alpha-collagen-related (Shc) effector, as the SH2-Shc domain prevented the GVBD and the ERK2 phosphorylation induced by FGF1. The importance of another signalling pathway involving phospholipase Cgamma (PLCgamma) was also investigated. The use of the PLCgamma inhibitory peptide, neomycin and the calcium chelator BAPTA-AM on oocytes expressing FGFR1 or the stimulation by PDGF BB of oocytes expressing PDGFR-FGFR1 mutated on the PLCgamma binding site, prevented GVBD and ERK2 phosphorylation. This study shows that the transduction cascade induced by the FGFR1-FGF1 interaction in Xenopus oocytes represents the sum of Ras-dependent and PLCgamma-dependent pathways. It emphasizes the role played by PI3K and Src and their connections with the Ras cascade in the FGFR1 signal transduction. PMID- 11012681 TI - Interaction of peroxisomes with microtubules. In vitro studies using a novel peroxisome-microtubule binding assay. AB - The association of membrane-bounded cell organelles to microtubules is crucial for determination of their shape, intracellular localization and translocation. We have shown previously the high affinity binding of peroxisomes to microtubules which appears to be of static nature as in vivo studies indicate that only a few peroxisomes move along the microtubular tracks. In order to characterize the interactions of peroxisomes with microtubules, we have developed a semiquantitative in vitro binding assay, which is based on the association of highly purified rat liver peroxisomes to microtubules coated onto microtiterplates. The binding was visualized by differential interference contrast and immunofluorescence using a confocal laser scanning microscope. The binding was concentration dependent and saturable, being affected by time, temperature, and pH. Addition of ATP or the motor proteins kinesin and dynein increased the binding capacity, while ATP-depletion or microtubule associated proteins (MAPs) decreased it. KCl treatment of peroxisomes reduced the binding, which was restored by dialyzed KCl-stripping eluate as well as by rat liver cytosol. The reconstituting effect of cytosol was abolished by its pretreatment with proteases or N-ethylmaleimide. Moreover, the treatment of peroxisomes with proteases or N-ethylmaleimide reduced their binding, which was not reversed by cytosol. These results suggest the involvement of a peroxisomal membrane protein and cytosolic factor(s) in the binding of peroxisomes to microtubules. This notion is supported by the observation that distinct subfractions of dialyzed KCl stripping eluate obtained by gel chromatography augmented the binding. Those subfractions, as well as purified peroxisome fractions, exhibited strong immunoreactivity with an antibody to cytoplasmic linker protein (CLIP)-115, revealing a 70-kDa polypeptide. Moreover, immunodepletion of KCl-stripping eluate and its subfractions with an antibody to the conserved microtubule binding domain of CLIPs, abolished their promoting effect on the binding, thus suggesting the involvement of a CLIP-related protein in the binding of peroxisomes to microtubules. PMID- 11012682 TI - Ether lipids in the cell membrane of Mycoplasma fermentans. AB - Two new ether lipids, 1-O-alkyl/alkenyl-2-O-acyl-glycero-3-phosphocholine and its lyso form, 1-O-alkyl/alkenyl-glycero-3-phosphocholine, were identified in the cell membrane of Mycoplasma fermentans using chemical analyses, GLC-MS, MALDI-TOF MS, and 1D and 2D NMR spectroscopy. The lipids are heterogeneous with respect to both acyl and alkyl/alkenyl residues. The acyl residues at position 2 of glycerol are hexadecanoyl and octadecanoyl in a molar ratio of 3.6 : 1 with a trace amount of octadecenoyl. The alkyl/alkenyl residues at position 1 of glycerol are hexadecyl (78%), octadecyl (7%), octadecenyl (14%), and hexadecenyl (traces). In the octadecenyl residue, the double bond has a cis configuration and is located at either position 1' (plasmalogen-type lipid) or 9' in a ratio approximately 1 : 1. This is the first report of the presence of alkyl and vinyl (alk-1'-enyl) ether lipids in the cell membrane of aerobically grown mycoplasmas. Lipids of this type have been found in some Gram-positive bacteria, thus supporting the hypothesized close taxonomical relationship of these bacteria to mycoplasmas. The ether lipids of M. fermentans are structurally similar to platelet activating factor; it was demonstrated that the 2-O-acetylated lyso form lipid can mimic platelet-activating factor activity in isolated perfused and ventilated rat lungs. PMID- 11012683 TI - Comparison of the substrate specificity of the human T-cell leukemia virus and human immunodeficiency virus proteinases. AB - Human T-cell leukemia virus type-1 (HTLV-1) is associated with a number of human diseases. Based on the therapeutic success of human immunodeficiency virus type 1 (HIV-1) PR inhibitors, the proteinase (PR) of HTLV-1 is a potential target for chemotherapy. To facilitate the design of potent inhibitors, the subsite specificity of HTLV-1 PR was characterized and compared to that of HIV-1 PR. Two sets of substrates were used that contained single amino-acid substitutions in peptides representing naturally occurring cleavage sites in HIV-1 and HTLV-1. The original HIV-1 matrix/capsid cleavage site substrate and most of its substituted peptides were not hydrolyzed by the HTLV-1 enzyme, except for those with hydrophobic residues at the P4 and P2 positions. On the other hand, most of the peptides representing the HTLV-1 capsid/nucleocapsid cleavage site were substrates of both enzymes. A large difference in the specificity of HTLV-1 and HIV-1 proteinases was demonstrated by kinetic measurements, particularly with regard to the S4 and S2 subsites, whereas the S1 subsite appeared to be more conserved. A molecular model of the HTLV-1 PR in complex with this substrate was built, based on the crystal structure of the S9 mutant of Rous sarcoma virus PR, in order to understand the molecular basis of the enzyme specificity. Based on the kinetics of shortened analogs of the HTLV-1 substrate and on analysis of the modeled complex of HTLV-1 PR with substrate, the substrate binding site of the HTLV-1 PR appeared to be more extended than that of HIV-1 PR. Kinetic results also suggested that the cleavage site between the capsid and nucleocapsid protein of HTLV-1 is evolutionarily optimized for rapid hydrolysis. PMID- 11012684 TI - Combinatorial mutagenesis and pseudorevertant analysis to characterize regions in loop E of the CP47 protein in Synechocystis sp. PCC 6803. AB - Deletion of the I265-F268 and T271-K277 regions in the large lumenally exposed loop of the CP47 protein are known to lead to a loss of photoautotrophic growth. Here, these regions have been investigated by combinatorial mutagenesis and pseudorevertant mapping. No single amino-acid residue in the I265-F268 region was found to be critical for function, but a large hydrophobic residue at position 267 and preferentially an aromatic residue at position 268 appeared to be required for photoautotrophic growth. Starting from an obligate photoheterotrophic mutant lacking the T271-K277 region, photoautotrophic pseudorevertants were generated with short in-frame tandem repeats near the site of the original deletion, partially or fully restoring the length of the original protein. These pseudorevertants were sensitive to oxygen indicating that the T271 K277 region may provide PS II stability and/or protection against oxygen dependent photoinactivation. Pseudorevertants with much improved photoautotrophic growth were also generated for one of the combinatorial mutants in the I265-F268 region. Surprisingly, the secondary mutations in these pseudorevertants mapped to the ferrochelatase gene. We speculate that the secondary mutation in ferrochelatase gene resulted in altered ferrochelatase activity. Decreased heme (phycobilin) biosynthesis and/or increased chlorophyll biosynthesis could then lead to improved PS II performance of the combinatorial CP47 mutant. PMID- 11012685 TI - Microheterogeneity of recombinant human phenylalanine hydroxylase as a result of nonenzymatic deamidations of labile amide containing amino acids. Effects on catalytic and stability properties. AB - The microheterogeneity of recombinant human phenylalanine hydroxylase (hPAH) was investigated by isoelectric focusing and 2D electrophoresis. When expressed in Escherichia coli four main components (denoted hPAH I-IV) of approximately 50 kDa were observed on long-term induction at 28-37 degrees C with isopropyl thio-beta D-galactoside (IPTG), differing in pI by about 0.1 pH unit. A similar type of microheterogeneity was observed when the enzyme was expressed (1 h at 37 degrees C) in an in vitro transcription-translation system, including both its nonphosphorylated and phosphorylated forms which were separated on the basis of a difference in mobility on SDS/PAGE. Experimental evidence is presented that the microheterogeneity is the result of nonenzymatic deamidations of labile amide containing amino acids. When expressed in E. coli at 28 degrees C, the percentage of the acidic forms of the enzyme subunit increased as a function of the induction time with IPTG, representing about 50% on 8 h induction. When the enzyme obtained after 2 h induction (containing mainly hPAH I) was incubated in vitro, its conversion to the acidic components (hPAH II-IV) revealed a pH and temperature dependence characteristic of a nonenzymatic deamidation of asparagine residues in proteins, with the release of ammonia. Comparing the microheterogeneity of the wild-type and a truncated form of the enzyme expressed in E. coli, it is concluded that the labile amide groups are located in the catalytic domain as defined by crystal structure analysis [Erlandsen, H., Fusetti, F., Martinez, A., Hough, E., Flatmark, T. & Stevens, R. C. (1997) Nat. Struct. Biol. 4, 995-1000]. It is further demonstrated that the progressive deamidations which occur in E. coli results in a threefold increase in the catalytic efficiency (Vmax/[S]0.5) of the enzyme and an increased susceptibility to limited tryptic proteolysis, characteristic of a partly activated enzyme. The results also suggest that deamidation may play a role in the long term regulation of the catalytic activity and the cellular turnover of this enzyme. PMID- 11012687 TI - The International Haemophilia Training Centres of the World Federation of Hemophilia: a 30-year review. PMID- 11012686 TI - Potency and selectivity of inhibition of cathepsin K, L and S by their respective propeptides. AB - The prodomains of several cysteine proteases of the papain family have been shown to be potent inhibitors of their parent enzymes. An increased interest in cysteine proteases inhibitors has been generated with potential therapeutic targets such as cathepsin K for osteoporosis and cathepsin S for immune modulation. The propeptides of cathepsin S, L and K were expressed as glutathione S-transferase-fusion proteins in Escherichia coli. The proteins were purified on glutathione affinity columns and the glutathione S-transferase was removed by thrombin cleavage. All three propeptides were tested for inhibitor potency and found to be selective within the cathepsin L subfamily (cathepsins K, L and S) compared with cathepsin B or papain. Inhibition of cathepsin K by either procathepsin K, L or S was time-dependent and occurred by an apparent one-step mechanism. The cathepsin K propeptide had a Ki of 3.6-6.3 nM for each of the three cathepsins K, L and S. The cathepsin L propeptide was at least a 240-fold selective inhibitor of cathepsin K (Ki = 0.27 nM) and cathepsin L (Ki = 0.12 nM) compared with cathepsin S (Ki = 65 nM). Interestingly, the cathepsin S propeptide was more selective for inhibition of cathepsin L (Ki = 0.46 nM) than cathepsin S (Ki = 7.6 nM) itself or cathepsin K (Ki = 7.0 nM). This is in sharp contrast to previously published data demonstrating that the cathepsin S propeptide is equipotent for inhibition of human cathepsin S and rat and paramecium cathepsin L [Maubach, G., Schilling, K., Rommerskirch, W., Wenz, I., Schultz, J. E., Weber, E. & Wiederanders, B. (1997), Eur J. Biochem. 250, 745-750]. These results demonstrate that limited selectivity of inhibition can be measured for the procathepsins K, L and S vs. the parent enzymes, but selective inhibition vs. cathepsin B and papain was obtained. PMID- 11012688 TI - Total joint arthroplasty in haemophilia. AB - In severely affected haemophilic patients arthropathy is a common problem which can lead to considerable pain and functional deficit. Surgical management, including total joint arthroplasty, can be undertaken if conservative management fails. A search of the literature showed that a number of studies describing the use of total knee arthroplasty (TKA) and total hip arthroplasty (THA) in haemophilia have been published, whereas shoulder, elbow and ankle arthroplasties are confined to case reports. This paper reviews the functional outcome of arthroplasty in the different joints, the postoperative and long-term complications, and the impact of HIV. Although complications are commonly described and the surgery is technically demanding, the results suggest that arthroplasty, particularly of the hip and knee, can be a valuable option in the management of severe haemophilic arthropathy. PMID- 11012689 TI - Porcine factor VIII provides clinical benefit to patients with high levels of inhibitors to human and porcine factor VIII. AB - The development of an inhibitor against Factor VIII is an important complication of haemophilia and occurs in approximately 31% of patients [1]. Despite various approaches to their management, the presence of these inhibitors remains a major cause of morbidity and mortality. Inhibitors may be low level [<5 Bethesda Units (BU)] or high level [>10 BU]. Low-level inhibitors usually remain low and do not respond to administered factor VIII with a rise of the inhibitor titre; high level inhibitors, in contrast, are characterized by a rapid rise in titre following treatment with factor VIII. Modalities of treatment of acute bleeding episodes in patients with inhibitors include 'overcoming' the inhibitor with very large doses of factor VIII, 'bypassing' the inhibitor blockade with products such as activated prothrombin complex concentrates or recombinant factor VIIa, 'removing' the inhibitor by plasmapheresis or immunoabsorption and 'repressing' it with immunosuppressive drugs and immune tolerance induction. An alternative approach is to use a porcine factor VIII concentrate which does not cross-react with the human factor VIII inhibitor. Following treatment with porcine factor VIII, functional factor VIII can be detected and the therapeutic levels correlate with cessation of bleeding. The use of porcine factor VIII may, however, result in the development of antiporcine factor VIII antibodies, limiting its use. Experience in patients with high-titre inhibitors indicates that porcine factor VIII therapy could be used in the presence of factor VIII inhibitors [2-11], including inhibitors to porcine factor VIII [5,7], and that haemostasis may be obtained in the absence of detectable levels of circulating factor VIII [7,11]. PMID- 11012690 TI - Protease inhibitor therapy and bleeding. AB - Shortly after the introduction of protease inhibitor drugs (PIs) for the treatment of human immunodeficiency virus infection an association between these drugs and an increased bleeding tendency in patients with hereditary bleeding disorders was observed. Not only do patients experience an increased bleed frequency in usual sites, but bleeds can also occur in unusual places such as the finger joints. Mucus membrane bleeding and haematuria are also common. Ritonavir appears to be associated with the highest risk of bleeding followed by indinavir. As yet there has not been enough experience with the newer PIs to assess fully their potential to induce increased bleeding, although nelfinavir seems to pose less of a risk than the original PIs. PI-associated bleeds tend to be more resistant to factor concentrate treatment and periods of prophylaxis may be required in individuals with frequent persistent bleeds. Patients continuing on PI therapy tend to develop a tolerance to this adverse effect with time. The mechanism of the bleeding tendency has not been elucidated. There is no consistent evidence of a disturbance of coagulation, fibrinolysis or platelet function which raises the possibility that PIs may exert a direct local effect on blood vessels. It is very important that this class-specific side-effect is recognized and understood by both treaters and patients. PMID- 11012691 TI - The economics of bleeding disorders. PMID- 11012693 TI - Combined factor V and VIII deficiency in Indian population. AB - The clinical and haematological heterogeneity in cases of the rare combined factor V and VIII deficiency has not been reported so far from India. Nine such cases belonging to five unrelated families have been analysed in the present study for the various haematological and clinical parameters. A very mild clinical presentation is seen in all these cases. The clinical manifestations, however, do not correlate with the plasma levels of these factors. PMID- 11012692 TI - Haemophilia care in central Scotland 1980-94. I. Demographic characteristics, hospital admissions and causes of death. AB - To estimate the resources required to manage patients with haemophilia in Scotland, we studied the demographic features, hospital admissions and causes of deaths for individuals with haemophilia A and B and von Willebrand disease, treated with blood products, during the period 1980-94 living in central Scotland. Data were obtained from 413 adults and children (93% ascertainment). The age distribution in 1980 revealed a paucity of individuals over 60 years but the number in this age group increased over the study period. Of those with haemophilia A and B, 63 and two respectively, became HIV positive. Hospital admissions rose from 103 to 168 per annum; the number of annual bed days utilized also increased, but there was marked annual fluctuation (790-1832). The rate of admission was greater for those with severe haemophilia A and this increased during the 15-year period mainly due to the clinical consequences of human immunodeficiency virus (HIV) and hepatitis C virus (HCV). The admission rate for haemophilia B was significantly lower than that for haemophilia A, and was similar for all degrees of severity of the disorder. Throughout the 15-year period the incidence of admissions for acute bleeds was constant, as was the average duration in hospital. For those with a factor VIII inhibitor, the rate of admission was about double the rate of those without an inhibitor, although the duration of hospital stay was similar for both groups. There were 61 deaths; the death rate increased during the study period principally due to HIV and HCV, and 12 patients died from haemorrhage. We conclude that: (i) the life expectancy for haemophiliacs in Scotland was generally increasing, although HIV and HCV caused increasing mortality and morbidity (as shown by the increase in hospital admissions); (ii) hospital bed usage for the treatment of acute bleeds continued to be required, but fluctuated greatly; and (iii) the clinical impression that haemophilia B is less clinically severe than haemophilia A is confirmed by objective data. The planning implications for haemophilia care in Scotland and similar countries are discussed. PMID- 11012694 TI - Seventeen years' experience with Autoplex/Autoplex T: evaluation of inpatients with severe haemophilia A and factor VIII inhibitors at a major haemophilia centre. AB - A retrospective case analysis of the use of Autoplex T (anti-inhibitor coagulant complex [heat treated]) at a major haemophilia treatment centre is described. Twenty-three haemophilia A patients with a history of high-titre (>10 BU), high response inhibitors were treated for a total of 54 bleeding episodes requiring hospital admission and/or surgical intervention. Effective or partially effective haemostatic control was achieved within 72 h in 94% of reported cases (effective 85%; partially effective 9%). The incidence of adverse reactions was low and there were no thrombotic complications. In bleeding episodes for which BU titres were determined within 6 months pre- and 6 months post-treatment, there was little tendency for Autoplex T to cause an anamnestic response (1 of 19 bleeding episodes). In the three invasive surgical procedures included in the case review, coverage with Autoplex T resulted in no postoperative complications, no bleeding at the surgical site, and no postoperative haematoma. The results support the efficacy and safety of Autoplex T in the treatment of bleeding episodes and haemostatic coverage of surgical procedures in haemophilia A patients with inhibitors. PMID- 11012695 TI - In vitro stability of recombinant human factor VIII (Recombinate). AB - Factor VIII (FVIII) is currently administered in diverse settings and by a range of methods, and it is important that the stability of specific FVIII preparations be documented for these varying uses. This study of Recombinate recombinant human FVIII (rhFVIII) evaluated: (i) thermostability; (ii) photostability; (iii) stability during simulated continuous infusion; and (iv) stability after dilution. This evaluation was conducted over a range of initial rhFVIII potencies and under differing conditions of temperature, light exposure, dilution and heparin usage. FVIII biological activity was measured by one-stage and chromogenic substrate assays. Microbiological assessment was also performed. Lyophilized rhFVIII was found to be highly thermostable, as evidenced by an energy of activation (Ea) of 16.2 kcal mol-1 and recovery of 99.3% of initial activity after incubation for 6 months at 40 degrees C and 93.8% at 60 degrees C for 2 months. No significant loss of activity could be detected after accelerated simulated natural daylight exposure of lyophilized rhFVIII, although partial activity loss was observed after similar exposure of reconstituted rhFVIII. Shielding in foil wrap effectively prevented such photodegradation of reconstituted rhFVIII. Based upon these results, exposure of lyophilized rhFVIII to sunlight is unlikely to affect stability adversely. Activity of reconstituted rhFVIII (22-106 IU mL-1) remained stable during simulated continuous infusion for 96 h at ambient (20-25 degrees C) and elevated (28-32 degrees C) temperature, and in the presence or absence of 1 U mL-1 heparin. After dilution of reconstituted rhFVIII, an immediate 14-42% loss of expected rhFVIII activity was observed depending upon diluent composition. Accordingly, potential partial loss of rhFVIII activity should be taken into account when dilution is being considered. rhFVIII remained sterile at least 96 h during simulated continuous infusion. rhFVIII is a robust preparation exhibiting biological stability under a wide array of clinically relevant conditions. PMID- 11012696 TI - Immune tolerance in a haemophilia A patient with high inhibitor using locally prepared lyophilized cryoprecipitate. AB - The immune tolerance in a 12-year-old haemophilia A patient was carried out at the Faculty of Medicine, Ramathibodi Hospital, Bangkok in 1998. His inhibitor titres ranged from 10 to 3450 Bethesda units (BU). He suffered from serious bleeding episodes requiring prolonged hospitalization and the disarticulation of the left knee joint. After obtaining informed consent, locally prepared lyophilized cryoprecipitate (LC), heat treated at 60 degrees C for 25 h, was given in a dose of 13 units kg-1 body weight of factor VIII three times per week. His inhibitor was increased from 15 to 580 BU within the first 4 weeks of immune tolerance. Finally, it was decreased to 40 BU in the 36th week. The only adverse effect was seroconversion of anti-hepatitis C virus after receiving 108 bottles of LC for 36 weeks. In conclusion, the locally prepared LC was able to control the bleeding episodes in a haemophilia A patient with high inhibitor. To our knowledge, this is the first reported case in Thailand. PMID- 11012697 TI - Immune tolerance for haemophilia patients with inhibitors: analysis of the western United States experience. The Tri-Regional Nursing Group. AB - The experience with immune tolerance (IT) induction therapy for haemophilia patients with inhibitors, instituted during 1990-97 at 17 haemophilia treatment centres in the western United States, was reviewed. IT was instituted in 104 of 139 (75%) of all identified haemophilia A and haemophilia B patients with inhibitors. Doses and schedules for IT varied but most patients were treated with a daily administration of 25-200 units kg-1 day-1. Successful immune tolerance (defined as Bethesda Unit titres < 1.0) was achieved in 57 of 81 (78%) patients who completed therapy. IT success was significantly greater for those patients with historic titres < 100 BU (P < 0.0003) and those with titres of < 10 BU at initiation of IT (P < 0.0001). IT success was uniformly achieved in 16 patients with low to moderate responding inhibitors. IT in infants < 2 years of age was problematic due to the presence of high titre inhibitors and complications of venous access. Frequent complications of IT included increased bleeding during intravenous access device insertion (20%) and infections associated with venous access devices (64%). IT is widely accepted as a treatment modality for haemophilia patients with inhibitors. Patient participation in IT regimens and management during therapy is facilitated greatly by the efforts of treatment centre nurse coordinators who are knowledgeable about haemophilia and its complications. Additional clinical trials will be necessary to define better initial dose, schedule, dose adjustment and success/failure criteria for IT. PMID- 11012698 TI - Proposal of a standard approach to dental extraction in haemophilia patients. A case-control study with good results. AB - We found no case-control studies on dental extraction in haemophilia patients in the literature even though the use of antifibrinolytic agents following a single infusion of factor VIII or IX has been accompanied by a lower number of bleeding complications in dental extractions. In this study we verified the incidence of bleeding complications after dental extraction in a group of 77 haemophilia patients. One hundred and eighty-four male patients requiring dental extraction represented the control group. All haemophilia patients received 20 mg kg-1 of tranexamic acid and a single infusion of factor VIII or IX to achieve a peak level about 30% of factor VIII or IX in vivo prior to dental extraction. Forty five of 98 (45.9%) dental extractions in haemophilia patients and 110 of 239 (46%) dental extractions in the control group were surgical ones. We registered two bleeding complications in the group of haemophilia patients (one late bleeding and one haematoma in the site of the anaesthetic injection) and one (a late bleeding) in the control group. The difference of bleeding complications in the two groups of patients were not statistically significant (P=0.2; OR 0.2; CI 0.01-2.22). The protocol proposed in this study, characterized by the feasibility and the number of haemorrhagic complications not different from normal population, make dental extractions in haemophilia patients possible on an out patient basis with a cost reduction for the community and minor discomfort for the patients. PMID- 11012699 TI - Participation in sports by Dutch persons with haemophilia. AB - Patients with haemophilia are now widely advised to participate in sports. However, no extensive data are available about their actual participation. A self administered questionnaire was used to investigate sports participation by persons with haemophilia in the Netherlands. All 293 questionnaires that were distributed have been returned; 217 were from people with severe haemophilia A or B (this is 40% of the severely affected Dutch population with haemophilia). Of this group, 155 (71%) participate in one or more sports: 66 swim, 36 cycle, 21 play tennis, 18 do fitness exercises, 16 ice skate, 10 ski and 10 play table tennis; in total 44 different sports are mentioned. Of a group of 16 persons with haemophilia who are unable to run, 10 (63%) participate in sport and of a group of 91 who can only run sometimes, 64 (70%) engage in sports. This shows that there are ample opportunities for persons with haemophilia to participate in sports, even if they are unable to run. Severely affected persons with haemophilia are as active as the general Dutch population; in fact, a higher proportion swims and/or cycles. In conclusion, sports participation in the Netherlands by persons with haemophilia is in accordance with World Federation of Hemophilia directives. PMID- 11012700 TI - Isotopic synovectomy with P-32 in paediatric patients with haemophilia. AB - Isotopic synovectomy is being proposed as an option in the treatment of patients with haemophilic arthropathy. We present our experience with 11 paediatric patients who underwent 17, P-32 isotopic synovectomies for chronic haemophilic arthropathy. P-32 was injected into the joint per protocol, approved by the institutional review board. All our patients were male. Nine were factor VIII and two were factor IX deficient. The following joints were treated: ankle (n=10 procedures), elbow (n=5) and knee (n=2). The first procedure was performed on December 1993. None were human immunodeficiency virus positive. Mean age at the first procedure was 10.8 years (range, 5.2-15.2 years). Mean pretreatment joint clinical scores using the World Federation of Hemophilia guidelines for the ankle was 5.5 (SD +/- 2.3), the elbow 4.2 (+/-2.5), and knee 5.5 (+/-3.5); the corresponding post-treatment scores were 2.6 (+/-2.0), 1.4 (+/-0.5) and 2.5 (+/ 3.5) respectively. Presynovectomy mean radiological scores using the Pettersson method were: ankle 1.8, elbow 1.8, and knee 1.5. A scoring system used in our centre for evaluating joints using magnetic resonance imaging (MRI) gave the following mean pretreatment scores: ankle 9.5, elbow 8.4, and knee 5.0. A marked decrease (an 80-100% decrease) in bleeding was seen in 13 of 17 procedures, and a moderate decrease (51-79% decrease) in two procedures, accounting for 85% reduction in bleeding into the target joints. The procedure was well tolerated and no untoward side-effects were noted as of May 1999, with a median follow-up of 40 months (range 19-65 months). None had any clinical evidence of cancer. Three patients had their joints retreated [elbow (one), ankle (two)]. These procedures were also well tolerated. In conclusion, in our study, isotopic synovectomy using P-32 appears to be feasible, safe and efficacious in the treatment of haemophilic arthropathy in paediatric patients who have been followed for a median of 40 months. As previously shown, MRI appears to give more detailed information about joint arthropathy than plain radiographs. PMID- 11012701 TI - Elbow joint, crutches and locomotion: special reference to persons with haemophilia. AB - A 52-year-old, trans-femoral amputee with haemophilia was hospitalized because of ambulatory problems arising from the osteo-arthropathic involvement of other major articulations. Reduced function in the upper limbs, caused by the effects of recurrent haemarthroses, resulted in additional problems concerning the usage of auxiliary ambulatory aids. The advantages and disadvantages of traditional and experimental crutches highlight the functional problems of ambulation in persons with concomitant upper limb pathologies. PMID- 11012702 TI - Synovectomy with rifampicine in haemophilic haemarthrosis. AB - The purpose of this paper was to assess the effectiveness of intra-articular injected rifampicine in haemophilic patients in order to achieve synovectomy by preventing repeated intra-articular bleeding. We have used this technique in haemophilic patients previously and reported our results on 13 cases [1]. Two hundred and fifty milligrams of rifampicine was injected into the elbow and ankle joints and 500 mg was injected into knee joints with 3-10 mL of lidocaine, depending on the joint size. The injections were repeated once a week for 7 weeks. Patients were only covered with antihaemophilic factor on the day of the injection at 30% above their coagulation level. We evaluated the results using two measures: subjective reports from the patient and objective assessment by the examiner. In the subjective reports the patient graded the results from their own perspective from 1 (poor) to 10 (excellent): 1-3, poor; 4-6, fair; 7-8, good; and 9-10, excellent. In the objective reports the grading was: excellent ('dry joint', full function, no haemarthrosis, no synovitis); good (clinical improvement, synovitis, reduction of haemarthroses, full function); fair synovitis (reduction of haemarthroses, no change in function); poor synovitis (persistent haemarthroses). This paper reports on the results of 38 patients with 39 joints with more that 3 years follow up, mean 1.8 years. There were 22 knees, nine elbows and eight ankles. Subjectively, there were excellent results in 21 joints (11 knees, six elbows and four ankles) good results in 15 joints (eight knees, three elbows and four ankles), fair results in two knees and a poor result in one knee. Objectively, results obtained were excellent in 20 joints (11 knees, six elbows and three ankles); good in 17 (nine knees, three elbows and five ankles); fair in one knee and poor in one knee. PMID- 11012703 TI - Intra-articular hyaluronic acid in the treatment of haemophilic arthropathy of the knee. Clinical, radiological and sonographical assessment. AB - Hyaluronic acid has been used successfully in the treatment of osteoarthritis since 1989. There is no experience in haemophiliacs in larger study groups. In a prospective study, 20 patients (21 knees) with haemophilic arthropathy of the knee received 20 mg hyaluronic acid by intra-articular injection for 5 consecutive weeks. Assessment included clinical scores, X-ray, magnetic resonance imaging (MRI) and biomechanical motion analysis before and 3 months after the first injection. The score of the WFH advisory committee and the Aichroth score for special evaluation of the knee were used. After an average period of 26 months, the World Federation of Hemophilia (WFH) score, the Aichroth score and the visual analogue scale were evaluated again. All patients had pain caused by their arthropathy, nine of them had positive antibodies to human immunodeficiency virus, and 15 had chronic hepatitis C. The mean WFH score was 8.1 points, the Petterson score was 7.3 points and the Aichroth score was 38 points (maximum 55 points). The WFH score decreased to 7.3 points, the Aichroth score improved to 40 points and the subjective assessment measured with a visual analogue scale improved from 5.3 to 3.7 points. No differences from MRI controls were detected. After 3 months, 14 of 20 patients improved subjectively, particularly in longer walking distance, stair-climbing or initial pain. These positive aspects were limited by arthropathy in adjacent joints. After 26 months 10 patients still are benefiting for up to 31 months follow-up. The average WFH score was 7.3 points, the Aichroth score 39 points, the visual analogue scale 4.0 points. We recommend hyaluronic acid for haemophilic arthropathy of the knee when regular conservative therapy has failed and operative treatment is not feasible. PMID- 11012704 TI - Participation in research: the economic advantages in a haemophilia research population. PMID- 11012705 TI - Sensitivity of the reaction time of the resonance thrombogram for factor VIII:C and factor IX deficiencies in the blood of dogs with haemophilia A or B. AB - The sensitivity of the reaction time of the resonance thrombogram (RTG-r) was examined for two instruments based on 105 canine samples with a reduced factor VIII:C activity and 26 samples with a reduced factor IX activity. These samples were taken from dogs suffering from haemophilia A (n=18) or B (n=3) before and at different times after the substitution therapy with fresh frozen plasma. The RTG r sensitivity was low; with reference to the total of all examined samples, it was between 0.39 and 0.65 for the factor VIII:C activity and between 0.46 and 0.62 for the factor IX activity, depending on the instrument and the testing procedure used. Low sensitivity was shown especially by the fact that even some of the samples with a factor VIII:C activity < 5% showed false negative results. Moreover, the correlation found between RTG-r and factor VIII:C activity (Spearman's rank correlation coefficient, r(S)=-0.556 to -0.700) as well as between factor IX activity and RTG-r (r(S)=-0.380 to -0.612) was only moderate. The low sensitivity of the RTG towards a reduced activity of the haemophilia factors VIII:C and IX contradicts the exclusive use of the RTG for monitoring haemophilic dogs as well as a global screening test for the haemostasis of dogs. PMID- 11012706 TI - Recombinant FVIIa (NovoSeven) continuous infusion and total hip replacement in patients with haemophilia and high titre of inhibitors to FVIII: experience of two cases. AB - In this report we describe our experience of total hip replacement in two patients with severe haemophilia A and high titres of inhibitors to FVIII. We used rFVIIa replacement therapy by continuous infusion to perform the surgery. The total amount of rFVIIa used in these two patients was very similar but the manner of administration was quite different. In our experience, it is an advantage to use a higher dose for shorter periods than a lower dose for a longer treatment period. Tranexamic acid by continuous infusion, and parallel saline infusion were useful for good haemostasis and avoided local thrombophlebitis in the side of rFVIIa infusion. PMID- 11012707 TI - Cardiac tamponade due to post-cardiac injury syndrome in a patient with severe haemophilia A and HIV-1 infection. AB - An 18-year-old man with severe haemophilia A (FVIII:C < 1%) and human immunodeficiency virus 1 (HIV-1) infection was admitted to the hospital with fever and chest pain for 7 days. Eight weeks prior to his admission he had an accident for which he underwent, at another hospital, clinical and laboratory examination that revealed bone fractures of the nose cavity, and he was given factor VIII concentrates for seven days due to nasal bleeding. On admission, chest roentgenogram showed a large cardiac silhouette and echocardiography confirmed the presence of a large quantity of pericardial fluid. A presumptive diagnosis of the post-cardiac injury syndrome was made and he was given anti inflammatory drugs plus infusion of recombinant factor VIII concentrate (35 units kg-1 b.i.d.). On the seventh day he exhibited cardiac tamponade for which he underwent subxiphoid pericardiotomy with drainage of approximately 1500 mL of bloody exudate. He had an uncomplicated recovery and 10 days later he left hospital. He was given a continuous prophylactic treatment of 15 units kg-1 of recombinant FVIII every 2 days for 6 months, and 30 months after this episode the patient is free of any symptom. PMID- 11012708 TI - Continuous infusion of recombinant activated factor VII during caesarean section delivery in a patient with congenital factor VII deficiency. AB - Recombinant activated factor VII (rFVIIa) can be used as an alternative therapy in patients with FVII deficiency. However, as the drug has a very short half life, continuous infusion could be a meaningful administration modality. We report the case of a 30-year-old woman with moderate FVII deficiency and human immunodeficiency virus infection who underwent a caesarean section delivery. She was treated with a continuous infusion of rFVIIa and did not suffer any bleeding complication. The continuous infusion of rFVIIa was a safe and effective therapeutic approach for our patient, maintaining her levels of FVII:C and avoiding bleeding during caesarean section and afterwards. PMID- 11012709 TI - Oral immune tolerance induction to factor VIII via breast milk, a possibility? PMID- 11012710 TI - Stress and transposable elements: co-evolution or useful parasites? AB - The activity of transposable elements can be induced by environmental and population factors and in particular by stresses in various organisms. A consequence of the increase in transposable element mobility is the creation of new genetic variability that can be useful in the face of stressful conditions. In this review, results supporting this hypothesis are presented and discussed. The main question is how stress induces the activity of transposable elements. We discuss hypotheses based upon the existence of promoters or fixation sites of transcription activators in the untranslated regions of transposable elements, similar to those found in regulatory regions of host defence genes. PMID- 11012711 TI - Candidate gene analysis of thyroid hormone receptors in metamorphosing vs. nonmetamorphosing salamanders. AB - We used two different experimental approaches to test the hypothesis that thyroid hormone receptor (TR) variation is associated with alternate life cycles modes in ambystomatid salamanders. In the first experiment, the inheritance of TRalpha and TRbeta genotypes was determined for metamorphic and non metamorphic offspring from backcrosses between Ambystoma mexicanum (an obligate metamorphic-failure species) and metamorphic F1 hybrids (A. mexicanum x A. tigrinum tigrinum). The segregation of TR genotype was independent of the expression of life cycle mode phenotype, and neither TR locus was linked to DNA markers that flank a major effect locus for life cycle mode. In the second experiment, a portion of the ligand-binding domain of TRalpha and TRbeta was cloned and sequenced for DNA samples from 14 different ambystomatid salamander populations, including obligate metamorphic, facultative metamorphic, and obligate metamorphic-failure taxa. Nucleotide sequence variation was found for both TRalpha and TRbeta, with several nonsynonomous substitutions that presumably code for nonconservative amino acid replacements. However, no general relationship was found between TR allelic variation and life cycle mode among populations or species. These data do not implicate TRs as candidate loci involved in the current maintenance or past evolution of alternate life cycle modes in members of the tiger salamander complex. PMID- 11012712 TI - The evolution of senescence under curtailed life span in laboratory populations of Musca domestica (the housefly). AB - The evolution of senescence may be explained by two different, but not mutually exclusive, genetic mechanisms. The antagonistic pleiotropy hypothesis predicts that senescence is a consequence of the fixation of alleles with pleiotropic effects favouring early life fitness, but bearing a cost in later life. The mutation-accumulation hypothesis attributes senescence to the accumulation of deleterious mutations with late-acting effects on fitness in mutation-selection balance. Experiments were carried out on the housefly, Musca domestica, in which large and small populations were maintained so that reproduction was limited to four or five days after reaching sexual maturity. Longevity declined significantly under the husbandry protocol and was largely the same in large and small populations; this is consistent with the random accumulation of deleterious alleles affecting longevity under curtailed life span, although laboratory adaptation cannot be ruled out entirely as a causal mechanism. An analysis of life-history data did not provide evidence for a trade-off between longevity and age at sexual maturity, developmental time, or dry body weight, but there was an apparent trade-off between longevity and early progeny production, in support of antagonistic pleiotropy. PMID- 11012713 TI - Genetic diversity in tetraploid populations of the endangered daisy Rutidosis leptorrhynchoides and implications for its conservation. AB - Polyploidy is an important variable in assessing the genetics of endangered plant species. Species consisting of populations with different chromosome numbers pose questions as to the mode of inheritance, relative variability status, population divergence and gene flow. The self-incompatible species Rutidosis leptorrhynchoides (Asteraceae) in south-eastern Australia is a good example. The remnant populations in the northern sector of the species range are diploid, whereas southern ones are either diploid or tetraploid. Allozyme analysis of the tetraploid populations showed tetrasomic inheritance confirming an autopolyploid genetic system, a modest increase in their allelic richness over diploid populations in the same region and a lack of genetic divergence. Conservation and replenishment strategies should take account of these genetic features of mixed ploidy. PMID- 11012714 TI - Transinfection of Wolbachia in the mediterranean flour moth, Ephestia kuehniella, by embryonic microinjection. AB - Wolbachia are maternally transmitted intracellular bacteria found in many arthropod species. They cause a reproductive incompatibility called cytoplasmic incompatibility (CI) in several hosts, including the Mediterranean flour moth, Ephestia kuehniella. Two strains of E. kuehniella, one from Yokohama city and the other from Tsuchiura city, express different levels of CI: the Yokohama strain expresses CI at a higher level than the Tsuchiura strain. In order to determine whether the difference of CI levels depends on Wolbachia or the host, we performed transinfection experiments in E. kuehniella by means of embryonic microinjection, and successfully transferred Wolbachia carried by the Yokohama strain into the Tsuchiura strain, from which the original Wolbachia had been removed by tetracycline treatment. The resulting transinfected strain expressed CI at a level near that of the Yokohama strain, suggesting that, in these strains of E. kuehniella, the level of CI is determined by Wolbachia rather than by the host. PMID- 11012715 TI - Altitudinal variation for B chromosome frequency in the characid fish Astyanax scabripinnis. AB - The analysis of three populations of the characid fish Astyanax scabripinnis located at different altitudes along the same stream has revealed the presence of a macro B chromosome in two high-altitude populations (1800 m and 1920 m) but its absence from a low-altitude population (700 m). Because the stream flows through very rugged mountains, with numerous falls ensuring that any gene flow occurs downstream only, the absence of B chromosomes from the low-altitude populations is best interpreted in the light of the parasitic theory of B chromosome evolution. Under this theory, we would expect B chromosomes to be more frequent where environmental conditions are more favourable for the species, because Bs are best tolerated there. The widespread presence of these B chromosomes in numerous Brazilian river headwaters, which are the preferred habitat for this species, support this possibility. PMID- 11012716 TI - Inter- and intraspecific sexual discrimination in the flour beetles Tribolium castaneum and Tribolium confusum. AB - In Tribolium castaneum (CS) and T. confusum (CF), intra- and interspecific rates of homosexual mounting have been measured. The intraspecific results are compatible with the hypothesis of both species being sexually indiscriminate. However, the CF intraspecific rates were very high (35%-53% of mountings were homosexual), suggesting a lower sexual attractiveness, or a stronger rejection to being mounted, of CF females relative to conspecific males. CS males discriminate between species but, in interspecific contacts, preferentially mounted CF males rather than CF females. CF males do not discriminate between species, but the loss of sexual attractiveness of CF females, or their rejection to being mounted, may act as a precopulatory isolation mechanism. PMID- 11012717 TI - Contact zone between chromosomal races of Mus musculus domesticus. 2. Fertility and segregation in laboratory-reared and wild mice heterozygous for multiple robertsonian rearrangements. AB - Litter size, anaphase I nondisjunction and X-Y dissociation at metaphase I were studied in homozygous and heterozygous house mice from a central Italian chromosomal hybrid zone between the CD (2n=22) race and the standard race (2n=40). We also observed the segregation of the two chromosomal forms (Robertsonian and non-Robertsonian) in male and female multiple heterozygotes from the karyotype of their offspring and chromosomal arm counts of metaphase II. Litter size was significantly reduced in the F1 hybrids, but there was no difference in litter size between male and female F1s. Fertility in wild mice decreased with increasing numbers of structural heterozygosities (0-5). Some metacentrics appear to be under meiotic drive but there was no rule as to which of the two forms was favoured in backcrosses. An original observation of a negative correlation between the length of metacentrics and transmission rate was described in hybrids. Slight cosegregation of chromosomes with a similar morphology was present in the progeny of males and females. These observations are discussed in relation to the stability of this hybrid zone through time. PMID- 11012718 TI - Identification of native and hybrid elms in Spain using isozyme gene markers. AB - Two elm taxa occur naturally in the Iberian Peninsula: the Field elm (Ulmus minor) and the Wych elm (U. glabra). In addition, a third taxon, the foreign Siberian elm (U. pumila), was probably introduced in the 16th century as an ornamental tree and has spread spontaneously throughout the Peninsula. The natural hybridization between U. minor and U. pumila produced new individuals whose morphological traits appear to be mixed. Ulmus pumila, as well as its hybrids, has a high resistance to Dutch elm disease (DED). For this reason, it is commonly used in breeding programmes. Extensive hybridization and the high mortality produced by the last DED epidemic have endangered the conservation of the native elm. In this study, isozyme analyses are used to characterize the taxa U. minor and U. pumila. Siberian elms from Spain and China are compared with the native U. minor. Siberian elm produces isozyme patterns that completely differentiate it from U. minor. Three loci are completely different between the species: 6Pgd2, Mdh1 and Prx2. Isozyme markers can also be used to distinguish native elms from the hybrids that have evolved for generations. PMID- 11012719 TI - Population genetic structure of Australian magpies: evidence for regional differences in juvenile dispersal behaviour. AB - Territorial group size in Australian magpies (Gymnorhina tibicen) ranges from monogamous pairs to groups of more than 20 individuals. It has been hypothesized that large territorial groups result from the retention of juveniles after a breeding effort. If this is true, local populations consisting of large groups are likely to exhibit the most genetic structure, because over time similar genotypes will tend to be confined to limited areas if juveniles are predominantly philopatric. The objective of the present study was to test this hypothesis using allozyme and mitochondrial DNA data to provide indirect estimates of regional gene flow (derived from hierarchical population subdivision analyses). These data were used in combination with estimates of group size to infer patterns of dispersal among magpie populations across mainland Australia. Territorial groups were significantly larger in the south-west compared to three eastern regions. Although inferred levels of gene flow were substantial for all four regions, a striking pattern emerged from both sets of genetic data: more differentiation was evident among populations in the south-western region than in any eastern region. We conclude that levels of juvenile dispersal influence group size in G. tibicen, because in the south-western region where groups were largest, populations were most genetically differentiated. Our results suggest that contrasting population genetic structures may develop within a single species as a result of differences in social system. PMID- 11012720 TI - Negative genetic correlation between traits of the Drosophila head, and interspecific divergence in head shape. AB - For morphological traits that are negatively correlated, the genetic correlation (rg) between them might strongly influence patterns of morphological divergence and shape. Here, the pattern of divergence between two sibling species of cactophilic Drosophila, D. buzzatii and D. koepferae, is examined for two traits that are known to be negatively correlated in other Drosophila species: face width (FW) and width of both eyes (EW). Head width (HW, the sum of FW and EW, i. e. the total width of the head capsule) was also examined. Genetic and phenotypic correlations were estimated in the laboratory G2 generation of a sample of wild D. buzzatii derived from a population where D. koepferae is not present. Phenotypic correlations were also estimated in D. buzzatii and D. koepferae from another, very different, population where the species are sympatric. Consistent with studies in other Drosophila species, rg was negative and significant for the correlation between FW and EW, and positive (but nonsignificant at a matrix-wide P-value of 0.05) for the correlations of HW with both FW and EW. This well defined correlation pattern was also consistent with the phenotypic correlations in both D. buzzatii and D. koepferae. No significant difference in these traits was detected between D. buzzatii populations, but head shape has diverged between D. buzzatii and D. koepferae. Specifically, the two negatively correlated traits, FW and EW, have evolved in opposite directions in these two species, with HW showing no significant interspecific difference. The overall picture of this divergence pattern shows a striking concordance with the present evidence of negative correlations between FW and EW, and is consistent with the notion of rg related constraints on the pattern of interspecific differentiation. PMID- 11012722 TI - Wolbachia segregation rate in Drosophila simulans naturally bi-infected cytoplasmic lineages. AB - Wolbachia are maternally transmitted endocellular bacteria infecting several arthropod species. In order to study Wolbachia segregation rate, Drosophila simulans females from an Indo-Pacific population (Seychelles) bi-infected by the two Wolbachia variants wHa and wNo were backcrossed to uninfected males in two conditions. In the first case, Seychelles males from a stock cured from its Wolbachia by tetracycline treatment were used. In the second case, the males came from a naturally uninfected Tunisian population. It was found that (i) the two Wolbachia variants can segregate, so that bi-infected females can produce a few offspring infected only by wHa or wNo. This occurs in both backcross conditions. (ii) Segregation leads more frequently to wHa than to wNo mono-infection. (iii) Wolbachia transmission is lower when the Seychelles genome is introgressed by the Tunisian genome, suggesting that host genomic factors might influence infection fate. PMID- 11012721 TI - Genetics of alpha-amanitin resistance in a natural population of Drosophila melanogaster. AB - The genetic basis of variation in resistance to natural toxins is of interest for both ecological and evolutionary genetics. The wide variety of larval resources used by Drosophila, both within and between species, makes flies an excellent system for studying causes and consequences of selection resulting from exposure to natural toxins associated with different resources. In this study we carry out a genetic analysis of alpha-amanitin resistance in a population sample of Drosophila melanogaster. Data from mapping crosses of chromosome III support a role for a naturally occurring polymorphism in a multidrug resistance gene (Mdr65A) in alpha-amanitin resistance. However, there are no amino acid differences between resistant and sensitive chromosomes at Mdr65A. Therefore, if Mdr65A mutants contribute to the difference between alpha-amanitin-resistant and alpha-amanitin-sensitive third chromosome lines, the underlying cause is a gene regulatory mutation. PMID- 11012723 TI - Physical mapping of rDNA genes, (TTAGGG)n telomeric sequence and other karyological features in two earthworms of the family Lumbricidae (Annelida: Oligochaeta). AB - A cytogenetical study was carried out on the chromosomes and nuclear DNA amounts of the terrestrial earthworms Octodrilus complanatus and Eisenia foetida (Annelida: Oligochaeta: Lumbricidae). Chromosomes were studied using Giemsa staining, banding methods and fluorescent in situ hybridization (FISH) with two repetitive DNA probes [rDNA and (TTAGGG)n]. rDNA FISH and silver staining consistently identified one chromosome pair per spread in both species. The telomeric sequence (TTAGGG)n hybridized with termini of all the chromosomes in both earthworms. Flow cytometry DNA assays showed that O. complanatus and E. foetida had different nuclear DNA contents (2C value=1.72 and=1.40 pg, respectively) but very similar base composition in their genomes. PMID- 11012724 TI - Chromosomal regions involved in hybrid performance and heterosis: their AFLP(R) based identification and practical use in prediction models. AB - In this paper, a novel approach towards the prediction of hybrid performance and heterosis is presented. Here, we describe an approach based on: (i) the assessment of associations between AFLP(R) markers and hybrid performance and specific combining ability (SCA) across a set of hybrids; and (ii) the assumption that the joint effect of genetic factors (loci) determined this way can be obtained by addition. Estimated gene effects for grain yield varied from additive, partial dominance to overdominance. This procedure was applied to 53 interheterotic hybrids out of a 13 by 13 half-diallel among maize inbreds, evaluated for grain yield. The hybrid value, representing the joint effect of the genetic factors, accounted for up to 62.4% of the variation in the hybrid performance observed, whereas the corresponding efficiency of the SCA model was 36.8%. Efficiency of the prediction for hybrid performance was evaluated by means of a cross-validation procedure for grain yield of (i) the 53 interheterotic hybrids and (ii) 16 hybrids partly related to the 13 by 13 half-diallel. Comparisons in prediction efficiency with the 'distance' model were made. Because the map position of the selected markers is known, putative quantitative trait loci (QTL) affecting grain yield, in terms of hybrid performance or heterosis, are identified. Some QTL of grain yield detected in the present study were located in the vicinity of loci reported earlier as having quantitative effects on grain yield. PMID- 11012725 TI - Assessment of genetic variability in a traditional cassava (Manihot esculenta Crantz) farming system, using AFLP markers. AB - Despite the urgent need to conserve domesticated plant genetic resources, and developing 'on farm' strategies of conservation, the impact of traditional farming practices and of their interaction with ecological factors on the structure and evolutionary dynamics of the genetic variability of crop populations has been little documented. We assessed the genetic variability of 31 varieties of cassava (M. esculenta Crantz) traditionally grown by Makushi Amerindians from Guyana, using AFLP markers. We used a sample of 38 varieties from an ex situ core collection as a reference. Accessions of wild cassava were also included. While clonality of the varieties was expected due to the vegetative propagation of cassava, 21 varieties presented intravarietal polymorphism. Among the varieties from a single site in Guyana, genetic diversity was the same as that in the accessions from the core collection. We suggest that incorporation of volunteer seedlings, produced by sexual reproduction, into the stock of varieties grown by the Makushi plays a major role in explaining both intravarietal polymorphism and the high level of genetic diversity. No correspondence was found between the structure of molecular diversity and variation observed for agronomic traits that are targets for selection by cultivators. As found in previous studies, all wild forms of cassava clustered together and separately from the cultivated varieties in a Neighbour-Joining dendrogram. These results are consistent with the hypothesis of a limited domestication event in a restricted area, followed by rapid diffusion of cultivated phenotypes and convergent evolution. Our results show that local varieties are an important source of genetic diversity, and highlight the importance of the interaction between human and ecological factors in the dynamics of this diversity. PMID- 11012726 TI - Intratetrad mating, heterozygosity, and the maintenance of deleterious alleles in Microbotryum violaceum (=Ustilago violacea). AB - The mating system of Microbotryum violaceum was investigated in populations that are polymorphic for mating-type bias, where individuals produce viable haploids of only one of the two required mating types. The cause of mating-type bias was identified as deleterious recessive alleles linked to mating type. Maintenance of the deleterious alleles was promoted by early conjugation among products of single meioses, such that the duration of the free-living haploid stage is minimized. This development was also observed in nonbiased isolates. As a consequence, the mating system tends toward mating within the tetrad, which might be expected to reduce heterozygosity. However, complete centromere linkage of mating type ensures conjugation between first division meiotic products, such that mating in M. violaceum is analogous to forms of meiotic parthenogenesis with first division restitution (i.e. automixis with central fusion). This fungus was used to test the prediction that this mating system would maintain heterozygosity in regions of the genome linked to centromeres. Therefore, populations were screened for additional heterozygous lethal recessive alleles linked to centromeres, and several examples were found. Furthermore, the occurrence of intratetrad mating in M. violaceum provides an explanation for low variation among individuals within populations, inconsistent estimates of outcrossing rates, low levels of mating between tetrads of one diploid individual, and high frequencies of haplo-lethal alleles in natural populations. PMID- 11012727 TI - F1 hybrid inviability in eucalyptus: the case of E. ovata x E. globulus. AB - The impact of inbreeding and hybridization on fitness was compared in the two co occurring forest tree species, Eucalyptus ovata and E. globulus, aimed at explaining the rarity of their hybrids in nature. The success of selfing, open pollination and outcrossing of both species and interspecific hybridization was monitored from seed-set to 10-year's growth in a field trial. There was a unilateral barrier to hybridization with seed-set obtained only with E. ovata females. The F1 hybrids exhibited reduced viability compared to intraspecific cross-types at virtually all stages of the life cycle and are clearly at a selective disadvantage compared with their open-pollinated E. ovata half-sibs with which they would directly compete in nature. Eucalyptus ovata and E. globulus overlap in their flowering time but the F1 hybrids flowered later with virtually no overlap with either species. The asynchronous flowering and reduced reproductive fitness of F1 hybrids would markedly limit the opportunity for advanced generation hybridization. Inbreeding similarly had a deleterious effect on the fitness of both species, and the F1 hybrids were most competitive with the E. ovata selfs. It is argued that changes in inbreeding levels of parental populations may be a key factor affecting the relative fitness of hybrids and their potential to impact on the pure species gene pool. Reduced fitness of the pure species through inbreeding may result in hybridization having its greatest evolutionary impact in small founder or relict populations. PMID- 11012728 TI - Population structure of the pestiferous moth Helicoverpa armigera in the eastern Mediterranean using RAPD analysis. AB - The genetic structure of the cotton bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), was studied in the eastern Mediterranean. Moths were sampled in six locations (five in Israel, and one in Turkey) and their genetic relationship was analysed using RAPD-PCR. Three 10-oligonucleotide primers revealed 84 presumptive polymorphic loci that were used to estimate population structure. Results reveal low level of genetic distances among Israeli and Turkish populations. The estimated values of FST and theta for the eastern Mediterranean populations were very low across all populations, indicating a high level of gene flow. Four distinct RAPD-product profile types were defined, and found in all Israeli and Turkish populations. Although no isolation by geographical distance was detected, topographical barriers may play a role in such isolation. PMID- 11012729 TI - Sex, linkage disequilibrium and patterns of parasitism in three species of cyclically parthenogenetic Daphnia (Cladocera: Crustacea). AB - To gain insight into genetic variation for resistance to parasites, this study assayed clonal variation in cyclically parthenogenetic Daphnia magna with respect to parasitic infection. Samples were collected from natural populations, and the allozyme phenotypes of infected hosts were compared to those of uninfected hosts. Differences between the clonal composition of the infected and uninfected class were evident in only two of 16 populations examined. This result stands in contrast to a study of species in the D. pulex and D. longispina species complexes, where clonal variation for infection was found in 12 of 25 populations (Little & Ebert, 1999). Considering all populations from both studies, associations between host genotype and infection were typically evident only in populations that showed low genotypic diversity and evidence of genetic disequilibria, with D. magna showing the least amount of disequilibria. This pattern is compatible with at least two possibly overlapping hypotheses. First, it may be that those populations lacking clonal variation for infection experienced weaker parasite-mediated selection. We can not rule out variation in selection pressure as an explanation, but found no evidence that the prevalence or intensity of parasitism differed either among species, or between those populations which showed clonal variation for infection and those that did not. Second, it could be that some populations, especially those of D. magna, have more frequent sexual recruitment than others. Sexual recombination breaks up gene combinations which are in linkage disequilibrium, and our method to detect clonal variation for resistance relies on linkage between genetic markers (allozymes) and resistance loci. Past work on Daphnia has shown that the level of sexual recruitment (which is in turn mediated by habitat permanency) is indeed commonly linked to the occurrence of genetic disequilibria. Our results may thus underestimate the prevalence of clonal variation for infection (especially for D. magna), because most of the populations analysed appeared to have high levels of sexual recruitment and therefore lacked the linkage disequilibrium that underlies associations between allozymes and susceptibility. PMID- 11012731 TI - Distribution of the transposable element mariner in anopheline mosquitoes. AB - We have surveyed the distribution of the transposable element mariner using PCR in 23 species of Anopheles mosquitoes, including all of the most important vectors of malaria in South-east Asia. Sequencing of the nine positive species revealed elements from the irritans, mauritiana and mellifera subfamilies. These are the first data showing the presence of three subfamilies of mariners in anophelines. The elements we encountered are likely to be inactive, based on the presence of multiple stop codons and/or frameshifts. PMID- 11012730 TI - Dispersion of the Cf-4 disease resistance gene in Lycopersicon germplasm. AB - In the past, numerous Cf genes have been reported in tomato (Lycopersicon esculentum Mill.) that confer resistance against leaf mould (Cladosporium fulvum Cke.). We are interested in genetic variation at Cf loci. Therefore, previously uncharacterized Cf genes were further analysed. Recognition of the AVR4 elicitor, DNA gel blot analysis, PCR analysis and sequencing of part of the Cf-4 locus showed that a large proportion of the accessions tested harboured the Cf-4 resistance gene. We concluded that despite differences in nomenclature, all these accessions harbour the same Cf-4 locus, probably introgressed from the same donor. The origin of the Cf-4 locus and the reasons for discrepancies with earlier reports are discussed. PMID- 11012732 TI - The genetic diversity of native, stocked and hybrid populations of marble trout in the Soca river, Slovenia. AB - The marble trout (Salmo marmoratus) is an endangered species in Slovenia (and in Italy, Croatia and Albania) because of hybridization resulting from intensive stocking. Eleven populations of trout from the Soca river basin (Slovenia) were analysed using 31 presumptive enzyme loci. Three European control samples representing the main European genotypes of brown trout were also analysed. This analysis confirmed the existence of extensive hybridization in the main river, resulting in an equal mixture of alleles belonging to marble trout, and Danubian and Atlantic brown trout (Salmo trutta). Despite the high level of introgression observed in the main Soca River, nearly pure populations of marble trout were found in the headwaters of five tributaries. The five stations with pure marble trout will be the basis of a rehabilitation programme. Hardy-Weinberg and linkage disequilibria were observed and hypotheses are discussed to explain this. A scenario for the colonization of the Adriatic region is proposed. PMID- 11012733 TI - The use of molecular markers to investigate the genetic structure of an oil palm breeding programme. AB - RFLP markers (40 probes covering 60% of the oil palm genome) have been used to assess genetic diversity within 54 palms of a specific oil palm breeding programme. A further 10 palms encompassing a broader range of origins were also included to provide a wider framework for comparative analysis. These palms represent a majority of the parents in a crossing programme which aims to combine the best features of African tenera germplasm with South-east Asian Deli dura material. Progeny from these crosses are planted in up to six sites on three continents, where the palms are being assessed in detail for yield components. A total of 157 RFLP bands were scored and the data analysed by calculating genetic distances according to Nei & Li, and by correspondence and cluster analysis. The relationships that emerge through this molecular analysis correspond well with known pedigree and provenance. For example, south-east Asian Deli dura material forms a cluster clearly distinct from AVROS pisifera gene pools, and a selection of African breeding material forms a further and broader grouping. The potential ability of markers to assist in oil palm breeding through examination of the genetic structure of crossing programmes is discussed, as is their value in parental selection to maintain residual diversity within specific breeding pools and use in evaluating the fidelity of breeding lineages. PMID- 11012734 TI - The effect of drought stress on inbreeding depression in four populations of the Mediterranean outcrossing plant Crepis sancta (Asteraceae). AB - The effect of physiological stress on the magnitude of inbreeding depression in plants has been the subject of few studies and is currently controversial because of contradictory results. We measured the inbreeding depression at three drought stress levels, precisely defined by a preliminary physiological experiment. We also tested the hypothesis that more highly self-compatible populations exhibit reduced inbreeding depression due to purging of deleterious mutations. The study was conducted on two populations of the annual and allogamous plant Crepis sancta collected from the French Mediterranean region and two other populations from marginal areas with various self-incompatibility levels. Drought stress did not increase inbreeding depression in terms of plant mortality but significantly increased the inbreeding depression for the date of first flowering, number of heads per plant and relative growth rate. The most self-fertile marginal population showed an absence of inbreeding depression in most of the measured traits indicating that purging could have taken place in this population. The three others populations showed relatively low and similar estimates of inbreeding depression (delta approximately 0.35). The relatively low values obtained compared to the results found in allogamous plants suggests that the absence of competition for C. sancta in our experiment probably underestimated the effects of inbreeding in natural populations where competition occurs. PMID- 11012735 TI - Hyperplastic polyps and DNA microsatellite unstable cancers of the colorectum. AB - Although the scientific and clinical rationale for classifying colorectal cancer according to mechanisms underlying genetic instability is well supported, little is known of the early morphogenesis of sporadic cancer showing high levels of DNA microsatellite instability (MSI-H). Evidence is accumulating that the traditional adenoma-carcinoma sequence may not apply to sporadic MSI-H colorectal cancer. The serrated pathway comprising hyperplastic polyps, mixed polyps and serrated adenomas may serve as the missing link. This review relates the recently described CpG island methylator phenotype (CIMP) to the serrated pathway. Two rate-limiting genetic steps may underlie the neoplastic pathway associated with CIMP. A transmembrane receptor expressed by pericryptal myofibroblasts (HPP1) may be implicated in the transition from normal to hyperplasia whereas inactivation of hMLH1 is responsible for the conversion of hyperplasia to dysplasia through loss of DNA mismatch repair. These mechanisms fit with clinical observations relating to sporadic MSI-H colorectal cancer, specifically proximal location, multiplicity, higher frequency among females and rapid evolution of early cancer. PMID- 11012736 TI - Expression of promyelocytic leukaemia protein in thyroid neoplasms. AB - AIMS: Promyelocytic leukaemia protein (PML) is an oncoprotein involved in the pathogenesis of acute promyelocytic leukaemia and is localized in distinct PML nuclear bodies. Our previous observation of overexpression of the PML in hormone sensitive normal tissues and malignant solid tumours, including the thyroid, led to this analysis of the PML expression in various thyroid neoplasms to characterize the importance of the PML in thyroid carcinogenesis. METHODS AND RESULTS: Immunohistochemistry was performed on paraffin-embedded tissue samples from 106 thyroid neoplasms after antigen retrieval by microwave. Immunoblotting was done with fresh frozen tissues in a few tumours. The PML was strongly expressed in all papillary carcinomas in diffuse or ball-shaped patterns. In the follicular neoplasms, the PML expression was variable, but there was no significant difference between adenomas and carcinomas. In the medullary carcinomas, the PML expression was either not detectable or was lower than in non neoplastic thyroids. Quantitatively different expression of the PML in various thyroid neoplasms was confirmed by immunoblotting. CONCLUSION: A significant difference of the PML expression according to the type of thyroid neoplasms suggests that the PML is important in papillary thyroid carcinomas, and furthermore, that PML expression may be used in differential diagnosis of thyroid neoplasms. PMID- 11012737 TI - Epstein-Barr virus in gastric carcinomas with lymphoid stroma. AB - AIMS: To clarify the relationship between the Epstein-Barr virus (EBV) and gastric carcinoma with lymphoid stroma (GCLS) in Koreans, and to characterize the EBV-positive GCLS. METHODS AND RESULTS: EBV infection was examined using EBER in situ hybridization and polymerase chain reaction in 45 cases of GCLS among Koreans, and in 292 consecutive cases of gastric carcinomas without lymphoid stroma (non-GCLS) as controls. EBV infection was found in 30 tumours (67%) of GCLS and 10 tumours (3.4%) of non-GCLS (P < 0.05). EBV-positive GCLS was more prevalent in males, poorly differentiated histological type and diffuse type in Lauren's classification, and tended to be located more in the middle third of the stomach than EBV-negative GCLS (P < 0.05). p53 overexpression was observed in 22% of GCLS (17% of EBV-positive GCLS and 33% of EBV-negative GCLS), and 34% of non GCLS (EBV-positive GCLS vs. non-GCLS: P = 0.056). The survival of the patient with GCLS was not correlated with EBV infection or p53 immunoexpression (follow up period: 11-97 months). CONCLUSIONS: GCLS in Koreans is strongly associated with EBV infection. The prognosis in GCLS is not dependent upon either the status of EBV infection or the status of p53 immunoexpression. PMID- 11012738 TI - Congenital gastrointestinal stromal tumour is morphologically indistinguishable from the adult form, but does not express CD117 and carries a favourable prognosis. AB - AIMS: The histological and immunohistochemical features of a congenital stromal tumour of the jejunum are compared with those of adult gastrointestinal stromal tumours (GIST). The literature concerning the diagnosis and prognosis of congenital small intestinal stromal tumours is reviewed. METHODS AND RESULTS: A term female infant presented with intestinal obstruction, from birth. Histology of a 15-mm jejunal nodule showed a predominantly spindle-cell tumour with epithelioid areas. There was a low mitotic count and mild nuclear pleomorphism, extensive necrosis and haemorrhage, and focal calcification. Immunohistochemically, tumour cells stained for muscle specific actin and vimentin. Staining for CD117 (c-kit), S100, desmin and CD34 was negative. The features were compared to those of seven adult cases: no morphological feature was specific to the congenital tumour, which was smaller than the adult cases. There were no ultrastructural features specific for a particular line of differentiation. Immunohistochemical staining patterns were similar, except for CD117, which was strongly positive in all adult tumours, but negative in the congenital tumour. CONCLUSIONS: This congenital jejunal stromal tumour morphologically resembled adult GIST, but lack of c-kit expression suggests that it is nosologically distinct. Despite the presence of histological features which would cause the tumour to be categorized as malignant in an adult, it is apparent from previous reports of congenital small intestinal stromal tumours that the prognosis is favourable. PMID- 11012739 TI - Secondary neoplasms of the male genital tract with different patterns of involvement in adults and children. AB - AIMS: The incidence, presentation and macroscopic and histological features of secondary neoplasms of the male genital tract are described with reference to their differential diagnosis. METHODS AND RESULTS: A retrospective study of cases from the Royal London Hospital yielded a total of 31 secondary neoplasms involving the testis: 14 at postmortem examination and 17 surgical specimens. Nine cases were leukaemias: six acute lymphoblastic and two acute myeloid leukaemias in children, and one chronic lymphocytic leukaemia in an adult. The commonest primary sites of metastases to the testis were prostate (six cases), stomach (five cases) and lung (three cases). There were two malignant melanomas and isolated examples of metastases from the adrenal gland (neuroblastoma), cerebellum (medulloblastoma), soft tissue (alveolar rhabdomyosarcoma), pancreas and rectum. Of the metastases from solid tumours, 12 involved the right testis only, three involved the left and four were bilateral. In seven of these cases there were multiple testicular nodules, in seven there was a single mass, and in the rest there was diffuse involvement. Secondary neoplasms represented 4.6% of all testicular neoplasms at autopsy, and 1.6% in surgical specimens. There were five secondary penile neoplasms: two each from the pancreas and prostate and one from the bladder. Two neoplasms metastatic to the spermatic cord, both from a gastric primary, were included in the series. CONCLUSIONS: Secondary neoplasms of the testis occur with a frequency comparable to other sites in the genitourinary tract, and metastases to the spermatic cord, epididymis, and penis, are rare in comparison. Disseminated neoplasms rarely present initially at this site and are histologically distinctive in adults, but in children they must be distinguished from primary small round blue cell tumours. PMID- 11012740 TI - Cutaneous perineurioma: a poorly recognized tumour often misdiagnosed as epithelioid histiocytoma. AB - AIMS: Eleven cases of cutaneous perineurioma were studied to further characterize the histological features of this entity. METHODS AND RESULTS: The histological and immunocytochemical features of 11 cases of cutaneous perineurioma were studied and detailed by two pathologists. Clinical data were obtained from the referring clinician. Seven patients were female and four were male with ages ranging from 19 to 67 years (median 41 years). Six lesions arose on the leg. Macroscopically lesions ranged from 4 mm to 14 mm in maximum diameter (median 7 mm). Diagnostic histological features included a nonencapsulated but sharply demarcated tumour with a dumbbell architecture. The tumour cells were spindle shaped with delicate inconspicuous cytoplasm and arranged in sheets, whorls or with a vague fascicular pattern. Epithelioid cells with moderate amounts of eosinophilic cytoplasm were frequently admixed with the spindle cells. One tumour had trabeculae of cells embedded within a dense collagenous stroma as described in sclerosing perineurioma. One case displayed a prominent myxoid stroma. Three further cases contained small foci of fibrosis or myxoid change suggesting a morphological spectrum exists in cutaneous perineurioma. Mitoses were exceptionally rare and necrosis and significant cytonuclear pleomorphism was not found. All tumours were epithelial membrane antigen (EMA) positive. Six cases showed focal positivity for factor XIIIa. Follow-up ranged from 5 months to 6 years (median 1 years). No tumour recurred or metastasized. CONCLUSIONS: The histological appearance of this tumour is broader than hitherto realized. Several cases in this series were misdiagnosed histologically and cutaneous perineuriomas may be more common than currently appreciated. PMID- 11012741 TI - Tamoxifen-associated postmenopausal adenomyosis exhibits stromal fibrosis, glandular dilatation and epithelial metaplasias. AB - AIMS: Adenomyosis is relatively rare in postmenopausal women but recent reports have described its occurrence in this age group in patients taking tamoxifen. This study describes the pathology of nine cases of adenomyosis in postmenopausal women who were taking this medication. METHODS AND RESULTS: The pathology of the nine tamoxifen-associated cases was compared to five cases of postmenopausal adenomyosis not associated with tamoxifen. Morphological features present within adenomyosis more often in those taking tamoxifen were cystic dilatation of glands (which sometimes resulted in grossly visible intramural cystic lesions), fibrosis of the stroma and various epithelial metaplasias. The proliferative activity within the adenomyosis, as determined by MIB1 staining, was higher in the tamoxifen group. CONCLUSIONS: The study supports an association between tamoxifen therapy and the presence of adenomyosis in postmenopausal women. The aforementioned morphological features are characteristic of polypoid and nonpolypoid surface endometrium associated with tamoxifen and their occurrence within the adenomyosis is likely to represent extension of the surface epithelial changes. Adenomyosis may be more common than is generally realized in women taking tamoxifen and may account for postmenopausal bleeding in these patients. PMID- 11012742 TI - Distribution of apoptotic cells and expression of apoptosis-related proteins along the intrahepatic biliary tree in normal and non-biliary diseased liver. AB - AIMS: The cell kinetics and homeostasis of biliary epithelial cells may be maintained differently along the biliary tree. In this study, the role of apoptosis in the maintenance and homeostasis of the intrahepatic biliary tree was evaluated. METHODS AND RESULTS: By counting apoptotic biliary cells and by immunostaining apoptosis-related proteins in normal liver, fatty liver, and those with acute viral hepatitis, chronic viral hepatitis, and hepatitis virus-related cirrhosis, it was found that the larger the intrahepatic bile ducts became, the more biliary epithelial cells underwent apoptosis. bcl-2, an inhibitor of apoptosis, was diffusely expressed in the interlobular bile ducts, but rarely detectable in the large and septal bile ducts. bcl-XL and mcl-1, inhibitors of apoptosis, and bax, a promoter of apoptosis, were diffusely expressed along the intrahepatic biliary tree. CD95, a direct inducer of apoptosis, was present in the large and septal bile ducts, but rarely in the interlobular bile ducts. CONCLUSION: The ratio of bax to bcl-2, as well as the expression of CD95 which differed at the interlobular versus large and septal bile ducts, may be responsible for the unique distribution of apoptotic biliary cells and involved in the homeostasis of the intrahepatic biliary tree. PMID- 11012743 TI - Changing patterns of histological subgroups during therapy of Ph1+ chronic myelogenous leukaemia. AB - AIMS: Bone marrow histopathology reveals a striking heterogeneity at diagnosis of Philadelphia chromosome positive (Ph1+) chronic myelogenous leukaemia (CML). Based on semiquantitative evaluations of the number of megakaryocytes and the content of fibres, various histological subtypes have been postulated. However, little information exists on whether these groups represent stable categories of the different classification systems and whether therapeutic regimes exert any influence on the putative shift of histological patterns. METHODS AND RESULTS: A retrospective clinicopathological study was performed on 396 bone marrow biopsies derived from 173 patients. There were at least two representative trephines taken at diagnosis and at median intervals of 16 months. Processing of the specimens involved immunostaining with CD61 (megakaryopoiesis) and Ret40f (erythropoiesis) and Gomori's silver impregnation technique. Based on morphometric analysis and in accordance with the general appearance of bone marrow histology three different histological subtypes were distinguished. These consisted of a granulocytic (51 patients), a predominantly megakaryocytic (73 patients) and a myelofibrotic pattern (49 patients). Follow-up biopsies revealed that a significant transition of histological groups occurred and that, independently of treatment modalities, the myelofibrotic category was associated with an unfavourable prognosis. Of the 124 patients without myelofibrosis at onset, 42% later transformed into the myelofibrotic subtype. However, these patients showed no prevalence of either a pre-existing granulocytic or megakaryocytic growth. Myelofibrotic changes were significantly associated with interferon (IFN) and busulfan (BU) therapy. On the other hand, a transition of a myelofibrotic into a nonfibrotic subtype was detectable in 17 of the 49 patients under study and related to hydroxyurea (HU) treatment. CONCLUSIONS: Histological classification systems of bone marrow features in CML do not represent stable patterns, but may be significantly altered by therapy, in particular IFN and HU. PMID- 11012744 TI - Lymphomatous features of aggressive NK cell leukaemia/lymphoma with massive necrosis, haemophagocytosis and EB virus infection. AB - AIMS: Aggressive natural killer (NK) cell leukaemia will be categorized as a distinct entity in the new WHO classification of malignant lymphomas. However, its non-leukaemic features remain unclear. We therefore investigated the morphological and immunophenotypic features of this lymphoma. METHODS AND RESULTS: Four cases with aggressive NK cell lymphoma were morphologically and immunohistochemically studied. All cases followed an aggressive course with death occurring within about 3 months of initial presentation. In these cases, the neoplastic cells disseminated throughout systemic lymph nodes and invaded various tissues and organs. The lymphoma cells were large cells showing nuclear irregularity and a pattern of sinusoidal invasion in lymph nodes. Apoptosis and coagulation necrosis were both frequently observed. Haemophagocytosis was observed in all cases. Neoplastic cells in paraffin-embedded tissue specimens from these patients had CD3(CD3epsilon)+ CD56(123C3)+ granzyme+ TIA-1+ EBERT+ CD43(MT1)- CD45RO(UCHL-1)- CD57(Leu7)- CD20(L26)- phenotypes. In the two cases where tissue was available for immunohistochemical study in frozen sections, neoplastic cells showed CD56(Leu19)+ perforin+ Fas ligand(FasL)+ CD2(Leu5b)- CD3(Leu4)- CD4(Leu3)- CD5(Leu1)- CD7(Leu9)- CD8(Leu2)- betaF1- TCRdelta1- phenotypes. CD16(Leu11b) was positive in one case. CONCLUSIONS: : Natural killer cell lymphomas appear to represent a non-leukaemic counterpart of aggressive natural killer cell leukaemia, a relationship similar to that in adult T-cell leukaemia/lymphoma. Awareness and diagnosis of this aggressive lymphoma is important because of its fulminant course. PMID- 11012746 TI - From this month's histopathology PMID- 11012745 TI - Commentary: aggressive NK cell lymphomas: insights into the spectrum of NK cell derived malignancies. PMID- 11012747 TI - Toll-like receptors: molecular mechanisms of the mammalian immune response. PMID- 11012748 TI - Pollution and the immune response: atopic diseases--are we too dirty or too clean? PMID- 11012749 TI - Characterization of CD4- CD8- CD3+ T-cell receptor-alphabeta+ T cells in murine cytomegalovirus infection. AB - In this study, we have investigated that after the intraperitoneal infection with murine cytomegalovirus (MCMV), the CD3+ CD4- CD8-(double negative; DN) T-cell receptor (TCR)alphabeta+ T cells increased in peritoneal cavity, liver and spleen in both resistant C57BL/6 and susceptible BALB/c mice. The total cellular population of these cells showed peak levels around day 5 after infection in all the three investigated organs and the following phenotypical and functional characteristics emerged. The peritoneal DN TCRalphabeta+ T cells expressed highly skewed TCRVbeta8 on day 5 after infection compared with the uninfected mice, but those in spleen and liver showed moderate and low skewed TCRVbeta8, respectively. The percentages of NK1.1+ DN TCRalphabeta+ T cells gradually decreased as did modulation of some of their activation markers consistent with an activated cell phenotype. The peritoneal DN TCRalphabeta+ T cells on day 5 after infection expressed the genes of interferon-gamma (IFN-gamma), tumour necrosis factor alpha, Eta-1 (early T-cell activation-1) and MCP-1 (monocyte chemoattractant protein 1) but lacked expression of interleukin-4 (IL-4). After in vitro stimulation with phorbol 12-myristate 13-acetate and calcium ionophore in the presence of Brefeldin A, higher frequencies of intracellular IFN-gamma+ DN TCRalphabeta+ T cells were detected in all three investigated organs of infected mice compared with those of uninfected mice. Stimulation of peritoneal DN TCRalphabeta+ T cells with plate-bound anti-TCRbeta monoclonal antibodies showed proliferation and also produced IFN-gamma but not IL-4. These results suggest that DN TCRalphabeta+ T cells were activated and may have an antiviral effect through producing IFN-gamma and some macrophage-activating factors during an early phase of MCMV infection. PMID- 11012750 TI - Disparate effects of phorbol esters, CD3 and the costimulatory receptors CD2 and CD28 on RANTES secretion by human T lymphocytes. AB - This study has examined the stimuli required for secretion of regulated upon activation, normal T-cell expressed, presumed secreted (RANTES) from T lymphocytes and found that stimuli such as phorbol 12-myristate 13-acetate (PMA), which are unable to support T-cell proliferation and interleukin-2 (IL-2) production, are nevertheless able to elicit strong secretion of RANTES. Conversely, stimuli such as CD2 and CD28 ligation, which are able to support T cell proliferation, are unable to elicit RANTES secretion. Coligation of CD3 and CD28 drives T-cell proliferation to a similar degree as CD2 and CD28 coligation, yet also supports modest RANTES secretion. Furthermore, CD28 ligation enhances the secretion of RANTES stimulated by PMA and this costimulatory effect is abrogated by the phosphoinositide 3-kinase inhibitor wortmannin. Our data also indicate that the observed effects of PMA on RANTES secretion are probably due to activation of protein kinase C (PKC) isoenzymes, since RANTES secretion was unaffected by the non-PKC activating 4alpha-phorbol ester, whilst the general PKC inhibitor Ro-32-0432 inhibits PMA-stimulated RANTES secretion. Moreover, the effect of PMA appears to be chemokine-specific because PMA was unable to increase secretion of the related CC chemokine MIP-1alpha. Under stimulation conditions where increases in [Ca2+]i occur (e.g. PMA plus ionomycin or CD3 plus CD28 ligation) RANTES secretion can be severely reduced compared with the levels observed in response to the phorbol ester PMA. Hence, whilst PKC-dependent pathways are sufficient for strong RANTES secretion, a calcium-dependent factor is activated which negatively regulates RANTES secretion. This correlates well with the observation that ligation of cytolytic T lymphocyte-associated antigen-4 (CTLA-4) (expression of which has been reported to be dependent on a sustained calcium signal), inhibits RANTES secretion induced by CD3/CD28, but has no effect on PMA-stimulated RANTES secretion. PMID- 11012751 TI - Activation of T-cell receptor-gammadelta+ cells in the intestinal epithelia of KN6 transgenic mice. AB - We analysed the properties of intraepithelial lymphocytes of small intestine (SI IEL) in KN6-transgenic (Tg) mice expressing cDNA of T-cell receptor (TCR) gammadelta specific for the T22b molecule. While most splenic Tg TCR-gammadelta+ cells from KN6-Tg mice with H-2d/d background (Tgd/d mice) were Thy-1+ CD8alpha- CD44dull+ CD45RB+ CD69-, Tg TCR-gammadelta+ cells in SI-IEL (Tg gammadelta-IEL) were heterogeneous in the expression of Thy-1, CD8alpha and CD44 molecules and predominantly CD45RB+ CD69+. Tg gammadelta-IEL exhibited a much reduced proliferative response to the antigen (irradiated H-2b splenocytes) than splenic Tg TCR-gammadelta+ cells; the CD44+ subset, but not the CD44- subset, in Tg gammadelta-IEL responded to the antigen. Furthermore, Tg gammadelta-IEL, but not splenic Tg TCR-gammadelta+ cells, displayed cytolytic activity whether they were prepared from conventional or germ-free KN6-Tg mice. Comparative analysis of young and aged KN6-Tg mice revealed that the proportion of CD44+ cells in Tg gammadelta-IEL increased but the proliferative response of Tg gammadelta-IEL to the antigen attenuated in association with ageing. Moreover, although Tg gammadelta-IEL from Tgb/d mice contained a higher proportion of CD44+ cells than Tgd/d mice, they did not respond to the antigen. These results demonstrate that Tg TCR-gammadelta+ cells lose the ability to recognize the antigen following activation in the intestinal epithelia. PMID- 11012752 TI - Poly-guanosine motifs costimulate antigen-reactive CD8 T cells while bacterial CpG-DNA affect T-cell activation via antigen-presenting cell-derived cytokines. AB - Pathogen-derived pattern recognition ligands like lipopolysaccharide (LPS) and bacterial cytidine-guanosine (CpG)-DNA not only activate dendritic cells and macrophages but are also mitogenic for B cells. Less clear are the claimed effects of CpG-DNA on T cells, which range from direct activation, costimulation, or indirect transient activation via antigen-presenting cell (APC)-derived interferon type I (IFN type I). Here we demonstrate that CpG-DNA sequence specifically triggers macrophages to produce IFN type I, interleukin (IL)-12, IL 6 and tumour necrosis factor (TNF), but lacks the ability to directly costimulate T cells. Strikingly, poly-guanosine (poly-G) extensions to CpG-containing oligonucleotides (ODN) abolished the macrophage stimulatory potential yet generated T-cell costimulatory activities. In fact, independently of CpG-motifs, poly-G-ODN displayed the ability to costimulate T cells. Costimulation was operative on CD8 T cells but not CD4 T cells. Poly-G-mediated costimulation resulted in IL-2-driven T-cell proliferation and induced cytolytic T cells. Overall the data imply that poly-G motifs costimulate antigen reactive CD8 T cells, while CpG-DNA motifs fail to do so but may affect T-cell activation via APC derived cytokines such as IFN type I. PMID- 11012753 TI - Cyclooxygenase-independent inhibition of dendritic cell maturation by aspirin. AB - When immature human myeloid dendritic cells were differentiated in vitro in the presence of aspirin, they were unable to stimulate T-cell proliferation. Aspirin and its major metabolite salicylate changed the surface marker phenotype of dendritic cells. The drugs particularly suppressed the levels of CD83 and the secreted p40 unit of interleukin-12 (IL-12), both markers of mature dendritic cells; 50% inhibitory concentration (IC50) values were 2.5 mM, a concentration more than 100 times greater than the concentration at mid-point inhibition (ID50) value for inhibition of prostaglandin synthesis. Concomitantly, the levels of CD14, a marker of monocytes/macrophages, increased above the levels found in immature dendritic cells. Cyclooxygenase inhibitors ketoprofen, indomethacin and NS-398 had no effect at concentrations more than a thousand-fold higher than their IC50 values. The effects were independent of the presence of prostaglandin E2 in the medium. Salicylates suppressed activation of the nuclear transcription factor kappaB, which regulates dendritic cell differentiation, but their effects on mature dendritic cells were negligible. Hence, aspirin inhibits dendritic cell function by inhibiting their terminal differentiation at concentrations achieved in the blood of patients chronically treated with high-dose aspirin. PMID- 11012754 TI - Calcitonin gene-related peptide inhibits lipopolysaccharide-induced interleukin 12 release from mouse peritoneal macrophages, mediated by the cAMP pathway. AB - Previously we showed that calcitonin gene-related peptide (CGRP), a neuropeptide, inhibited lipopolysaccharide (LPS)-induced tumour necrosis factor-alpha (TNF alpha) production and increased interleukin (IL)-6 release at low concentrations via activation of the cAMP pathway in mouse peritoneal macrophages (Mphi). In this study we examined whether CGRP could modulate IL-12 release from mouse peritoneal Mphi, and if so, what signal transduction pathway was involved. Mphi were obtained from the peritoneal exudate of male BALB/c mice. The cells were plated on culture dishes at a density of 5 x 105 cells per well and allowed to adhere for 2 hr. After incubation for 24 hr, the Mphi were cultured with 0.1 microg/ml of LPS, alone or together with CGRP (1-1000 nM) for 24 hr. The amount of IL-12 in the cell medium was measured by enzyme-linked immunosorbent assay (ELISA). The results showed that CGRP attenuated LPS-induced IL-12 release in a concentration-dependent manner. Production of IL-12 was decreased from 95.9+/-4.6 to 73.4+/-5.7 pg/ml by 100 nM CGRP. The two cAMP phosphodiesterase (PDE) inhibitors, 3-isobutyl-1-methyl-xanthine (IBMX) and rolipram, significantly potentiated the CGRP response, and the level of IL-12 was further decreased by 28% and 47%, respectively. However, CGRP had no effect on IL-12 production from unstimulated Mphi. The LPS-induced IL-12 release from Mphi could also be reduced by forskolin, an activator of adenylate cyclase, and 8-Br-cAMP, an analogue of cAMP. Using the reverse transcription-polymerase chain reaction (RT-PCR), we found that CGRP also decreased the LPS-induced IL-12 p40 mRNA levels. Furthermore, pretreatment with H89 (0.1 microM or 1 microM), an inhibitor of cAMP dependent protein kinase, diminished CGRP effects, IL-12 production and gene expression. These data suggest that LPS-induced IL-12 release and gene expression were attenuated by CGRP via an activated cAMP-protein kinase A (PKA) pathway in mouse peritoneal Mphi. PMID- 11012756 TI - Analysis of the mechanism for extracellular processing in the presentation of human immunodeficiency virus-1 envelope protein-derived peptide to epitope specific cytotoxic T lymphocytes. AB - An immunodominant epitope of human immunodeficiency virus-1 (HIV-1) gp160 recognized by Dd class I major histocompatibility complex (MHC) molecule restricted, CD8+ cytotoxic T lymphocytes (CTL) was originally identified as a peptide composed of 15 amino acids (P18IIIB: RIQRGPGRAFVTIGK). However, further study has indicated that a 10-mer peptide, I-10 (RGPGRAFVTI), within P18IIIB is the minimal-sized epitope and the trimming step(s) of two carboxyl terminal amino acids (GK) is essential to produce I-10 from P18IIIB. In the processing, angiotensin-1-converting enzyme (ACE), found in sera, plays a central role in generating I-10. Target cells could be sensitized with I-10 under conditions where ACE activity in the sera was abrogated. In contrast, in the case of P18IIIB, requiring further processing to delete the C-terminus of two amino acids in order to act, sensitization of target cells was completely abrogated under the conditions. Pretreatment of target cells with brefeldin A (BFA), preventing the presentation of endogenous antigens from the class I MHC molecule pathway, did not inhibit the presentation of P18IIIB. Moreover, glutaraldehyde-fixed cells, which can not process native protein, though they could present the exogenously added peptides, were also sensitized by P18IIIB. These results clearly demonstrate that the fine processing to produce I-10 occurred in the extracellular milieu. Furthermore, our result suggests that the longer P18IIIB can bind to the class I molecules on the cell surface, and then be trimmed by ACE while it is bound. The mechanisms behind the extracellular processing outlined in this paper will offer important information for designing peptide-based vaccines to elicit MHC molecule-restricted effectors. PMID- 11012755 TI - In vitro treatment of human transforming growth factor-beta1-treated monocyte derived dendritic cells with haptens can induce the phenotypic and functional changes similar to epidermal Langerhans cells in the initiation phase of allergic contact sensitivity reaction. AB - Human monocyte-derived dendritic cells (MoDCs) obtained from peripheral blood monocytes (PBMC) cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) can be activated in vitro by a variety of simple chemicals such as haptens and several metals. Recently, it has been demonstrated that transforming growth factor-beta1 (TGF-beta1) can induce further differentiation of MoDCs to the cells that share some characteristics with epidermal Langerhans cells, i.e. they contain Birbeck granules and express E cadherin. In this study, using such TGF-beta1-treated dendritic cells (TGF-beta1+ DCs), we examined the in vitro effects of representative haptens, i.e. NiCl2 and dinitrochlorobenzene (DNCB), on their phenotypic and functional characteristics, comparing with those reported in vivo in epidermal Langerhans cells during the sensitization phase of a contact sensitivity reaction. Treatment of TGF-beta1+ DCs with NiCl2 increased their expression of the molecules related to antigen presentation such as CD86, major histocompatibility complex class I and class II, and CD83, although weakly, in addition to that of those essential for their migration to the regional lymph nodes, such as CD49e, CD44 and its variant 6, while it down-regulated the expression of the molecules required for homing to the skin and staying in the epidermis, such as cutaneous leucocyte antigen (CLA) and E-cadherin. It also increased the production of tumour necrosis factor-alpha, but not that of IL-1beta or IL-12. DNCB also increased their CD86 expression and down-regulated E-cadherin and CLA, but did not affect other phenotypic changes that were observed in TGF-beta1+ DCs treated with NiCl2. TGF-beta1+ DCs treated with either NiCl2 or DNCB increased their allogeneic T-cell stimulatory function. In addition, reverse transcribed polymerase chain reaction revealed augmented expression of chemokine receptor 7 mRNA by TGF-beta1+ DCs when treated with either NiCl2 or DNCB. Moreover, consistent with this data, TGF-beta1+ DCs treated with these chemicals chemotactically responded to macrophage inflammatory protein 3beta. These data suggest the possibility that TGF-beta1+ DCs present a good in vitro model to study the biology of epidermal Langerhans cells. PMID- 11012757 TI - The contribution of both oxygen and nitrogen intermediates to the intracellular killing mechanisms of C1q-opsonized Listeria monocytogenes by the macrophage-like IC-21 cell line. AB - Listeria monocytogenes is a facultative intracellular pathogen which is internalized by host mammalian cells upon binding to their surface. Further listerial growth occurs in the cytosol after escape from the phagosomal-endosomal compartment. We have previously reported that C1q is able to potentiate L. monocytogenes phagocytosis upon bacterial opsonization by ingestion through C1q binding structures. In this report, we analysed the post-phagocytic events upon internalization of C1q-opsonized L. monocytogenes and found an induction of macrophage (Mphi)-like IC-21 cell bactericidal mechanisms displayed by the production of oxygen and nitrogen metabolites. Both types of molecules are effective in L. monocytogenes killing. Further analysis of the cellular responses promoted by interaction of C1q with its surface binding structures, leads us to consider C1q as a collaborative molecule involved in Mphi activation. Upon interaction with surface binding structures, C1q was able to trigger and/or amplify the production of reactive oxygen and nitrogen intermediates induced by stimuli such as interferon-gamma and L. monocytogenes phagocytosis. PMID- 11012758 TI - Reversal of ultraviolet radiation-induced immune suppression by recombinant interleukin-12: suppression of cytokine production. AB - Exposure to ultraviolet (UV) radiation, a complete carcinogen, suppresses the immune response. Data from a number of laboratories have indicated that one consequence of UV exposure is suppressed T helper type 1 (Th1) cell function with normal Th2 cell activation, resulting in a shift to a Th2-like phenotype. The reversal of UV-induced immune suppression and tolerance induction by recombinant interleukin-12 (rIL-12) supports this observation. The focus of this study was to determine the mechanism(s) by which rIL-12 reverses UV-induced immune suppression. Two possibilities were considered: up-regulation of interferon-gamma (IFN-gamma) secretion by rIL-12 and suppression of UV-induced cytokine secretion by rIL-12. To our surprise we found that the ability of rIL-12 to overcome UV induced immune suppression was independent of its ability to up-regulate IFN gamma secretion. Rather, rIL-12 suppressed the production of cytokines that are known to be important in UV-induced immune suppression. Injecting UV-irradiated mice with rIL-12, or adding rIL-12 to UV-irradiated keratinocyte cultures suppressed IL-10 secretion, in part by affecting the transcription of the IL-10 gene. Furthermore, we found that rIL-12 suppressed UV-induced tumour necrosis factor-alpha (TNF-alpha) production. Because IL-10 is involved in the UV-induced suppression of delayed-type hypersensitivity and TNF-alpha in the UV-induced suppression of contact allergy, these findings provide a mechanism to explain how rIL-12 overcomes UV-induced immune suppression in these related but different immune reactions. In addition, they suggest a novel mechanism by which rIL-12 alters immune reactivity, direct suppression of cytokine secretion induced by UV radiation. PMID- 11012759 TI - Alteration in the responsiveness to tumour necrosis factor-alpha is crucial for maximal expression of monocyte chemoattractant protein-1 in human neutrophils. AB - We previously reported delayed expression of monocyte chemoattractant protein-1 (MCP-1) in human neutrophils cultured with a cytokine-rich crude supernatant of phytohaemagglutinin-stimulated peripheral blood mononuclear cells (PHA-sup). Tumour necrosis factor-alpha (TNF-alpha) contained in the PHA-sup played a key role in this event, but there appeared to be another factor(s) in the same supernatant that co-operated with TNF-alpha for maximal MCP-1 expression. In the present study, we reduced TNF-alpha concentrations in the PHA-sup to minimal levels using anti-TNF-alpha affinity columns (TNF-depleted-sup) and investigated the co-operation between TNF-alpha and TNF-depleted-sup. Nine hours of preincubation with TNF-depleted-sup altered the responsiveness of neutrophils to TNF-alpha and enabled TNF-alpha to increase the level of MCP-1 expression to a maximal level within 4 hr. The priming effect was not due to the increased expression of cell-surface TNF receptors. However, the activation of primed cells by TNF-alpha was clearly through TNF receptor-p55. Finally, the activity in the TNF-depleted-sup that co-operated with TNF-alpha was eluted at 60 000 MW on high performance liquid chromatography-gel filtration. Thus, delayed neutrophil expression of MCP-1 is regulated by a cytokine-dependent mechanism that induces neutrophils to enter a 'mature' stage. PMID- 11012760 TI - Structure/function studies of human decay-accelerating factor. AB - The decay-accelerating factor (DAF) contains four complement control protein repeats (CCPs) with a single N-linked glycan positioned between CCPs 1 and 2. In previous studies we found that the classical pathway regulatory activity of DAF resides in CCPs 2 and 3 while its alternative pathway regulatory activity resides in CCPs 2, 3 and 4. Molecular modelling of the protein predicted that a positively charged surface area on CCPs 2 and 3 (including KKK125-127) and nearby exposed hydrophobic residues (L147F148) on CCP3 may function as ligand-binding sites. To assess the roles of the N-linked glycan and the above two sets of amino acids in the function of DAF, we mutated N61 to Q, KKK125-127 to TTT and L147F148 to SS. Following expression of the mutated cDNAs in Chinese hamster ovary cells, the glycosylphosphatidylinositol (GPI)-anchored mutant proteins were affinity purified and their functions were assessed. In initial assays, the proteins were incorporated into sheep and rabbit erythrocytes and the effects of the mutations on regulation of classical and alternative C3 convertase activity were quantified by measuring C3b deposition. Since DAF also functions on C5 convertases, comparative haemolytic assays of cells bearing each mutant protein were performed. Finally, to establish if spatial orientation between DAF and the convertases on the cell surface played any role in the observed effects, fluid phase C3a generation assays were performed. All three assays gave equivalent results and showed that the N-linked glycan of DAF is not involved in its regulatory function; that L147F148 in a hydrophobic area of CCP3 is essential in both classical and alternative pathway C3 convertase regulation; and that KKK125 127 in the positively charged pocket between CCPs 2 and 3 is necessary for the regulatory activity of DAF on the alternative pathway C3 convertase but plays a lesser role in its activity on the classical pathway enzyme. PMID- 11012761 TI - Immunoglobulin E antibodies of atopic individuals exhibit a broad usage of VH gene families. AB - The term 'atopy' describes the genetically determined tendency to mount immunoglobulin E (IgE) antibody responses against per se harmless antigens (allergens). In this study we investigated the usage of VH families in the formation of IgE antibodies in 10 patients suffering from mucosal and/or skin manifestations of atopy. IgE antibody reactivities to exogenous allergen sources as well as to autoallergens were determined and, by immunoabsorption, it was demonstrated that allergen-specific IgE accounted for most of the total serum IgE levels in these patients. Using primers with specificity for the VH1-6 gene families and a primer specific for the first constant region of human IgE, cDNAs coding for IgE heavy chain fragments were amplified using the reverse transcription-polymerase chain reaction (RT-PCR) from peripheral blood lymphocytes of the 10 atopic individuals. Hybridization of the heavy chain encoding cDNAs with an IgE-specific internal oligonucleotide probe revealed a broad usage of all VH-gene families in the atopic individuals. The spectrum of VH families used in a given atopic individual was neither associated with the type or severity of clinical symptoms nor with the number of allergens recognized. The fact that allergen-specific IgE antibodies in atopic individuals originate from a broad variety of B cells thus reflects the activation of multiple B-cell clones during allergen sensitization. This finding should be borne in mind if therapeutic strategies for Type I allergy are considered that aim at a clonal elimination of allergen-specific B cells. PMID- 11012762 TI - Feline leukaemia virus: protective immunity is mediated by virus-specific cytotoxic T lymphocytes. AB - Feline leukaemia virus (FeLV) nucleic acid vaccination of domestic cats affords protection against viraemia and the development of latency without inducing antiviral antibodies.1 To determine the contribution of cell-mediated immunity to the control of virus replication and clearance from the host, FeLV-specific cytotoxic T lymphocyte (CTL) responses were compared in vaccine-protected, transiently viraemic, and persistently viraemic cats. Vaccinal immunity was associated with the detection of higher levels of virus-specific effector CTL in the peripheral blood and lymphoid organs to FeLV Gag/Pro and Env antigens than those observed in unvaccinated control, persistently viraemic cats (P<0.001). Likewise, higher levels of virus-specific CTLs were also observed in transiently viraemic cats which recovered following exposure to FeLV. In cats that controlled their infection, recognition of Gag/Pro antigens was significantly higher than the recognition of Env antigens. This is the first report highlighting the very significant role that virus-specific CTL have in determining the outcome of FeLV infection in either vaccinated cats or cats recovering naturally from FeLV exposure. PMID- 11012763 TI - Immunity to vaginal herpes simplex virus-2 infection in B-cell knockout mice. AB - We investigated the involvement of antibody in protection against vaginal herpes simplex virus type-2 (HSV-2) infection by comparing intact and B-cell knockout (KO) mice. Vaginal immunization of intact mice with attenuated HSV-2 markedly reduced an HSV-2 challenge infection in the vagina. In contrast, immunization of B-cell KO mice produced less immunity against the challenge infection and that immunity occurred in a different pattern. At 20 hr after challenge, immunostaining of virus proteins in the vaginal epithelium and shed virus protein titres in the vaginal secretions were not significantly different between immunized and non-immunized B-cell KO mice and were much greater than in immunized intact mice. At 48 hr after challenge, the vaginal infection in immunized B-cell KO mice was markedly less than at 20 hr but remained approximately sevenfold higher than in intact mice. This pattern of challenge infection in the vagina indicates that B cells, and probably the antibody derived from them, provided significant protection against reinfection in intact mice, especially during the first 20 hr after challenge, while other effector mechanisms became important between 20 and 48 hr after challenge. To determine whether T-cell immunity in immunized B-cell KO mice was equal to that in intact mice, we assessed interferon-gamma (IFN-gamma) secretion by memory T cells in vivo in the vagina at 20 hr after challenge. We found no significant differences in the up-regulation of major histocompatibility complex (MHC) class II antigens in the epithelium, up-regulation of vascular cell adhesion molecule-1 (VCAM-1) in vascular endothelium, or recruitment of T cells to the mucosa, indicating that the memory T-cell response to virus challenge was the same in intact and B-cell KO mice. PMID- 11012764 TI - T lymphocytes from granulocyte colony-stimulating factor-/- mice produce large quantities of interferon-gamma in a chronic infection model. AB - Little is known about the role of granulocyte colony-stimulating factor (G-CSF) in the response to chronic bacterial infections. To address this we infected G CSF knock out (G-CSF-/-) mice with Mycobacterium avium. Infection was not exacerbated in G-CSF-/- mice despite a deficiency in the total bone marrow cells, colony-forming haemopoietic cells, granulocytes and monocyte precursors in the bone marrow. Peritoneal cells from G-CSF-/- produced less nitric oxide (NO) upon culture in vitro with antigen than did wild-type (WT) cells. Unexpectedly, T cells from infected G-CSF-/- mice were able to produce significantly more interferon-gamma (IFN-gamma) than the wild type (WT) controls. T cells from G-CSF /- mice still produced more IFN-gamma even when in vitro NO production was inhibited, while enzyme-linked immunospot assay (ELISPOT) assays showed more IFN gamma-producing cells in the G-CSF-/- mice. This was confirmed by intracellular cytokine staining (ICCS), which showed that there were more IFN-gamma producing T cells in vivo in the G-CSF-/- than the WT controls following M. avium infection. It is possible that a deficit of NO in vivo allows T cells to develop a higher IFN-gamma-producing phenotype. Thus we show a novel relationship between G-CSF and IFN-gamma production by T cells revealed in this chronic bacterial infection model. PMID- 11012765 TI - Interleukin-8 fails to induce human immunodeficiency virus-1 expression in chronically infected promonocytic U1 cells but differentially modulates induction by proinflammatory cytokines. AB - This study addresses the role of interleukin (IL)-8, a CXC-chemokine, the level of which is reported to be raised in the peripheral circulation of human immunodeficiency virus-1 (HIV-1)-infected individuals, during the induction of HIV-1 expression from latency and during cytokine-mediated HIV-1 up-regulation. IL-8 at the higher concentrations tested (> or = 100 ng/ml) was unable to induce HIV-1 expression in the chronically infected promonocytic U1 cell line, as measured by p24 antigen enzyme-linked immunosorbent assay (ELISA), whereas at lower concentrations of 1 and 10 ng/ml, constitutive HIV-1 expression was only marginally reduced. HIV-1 replication in acutely infected U937 cells was also significantly reduced by IL-8. The potent up-regulation of HIV-1 expression in U1 cells by tumour necrosis factor-alpha (TNF-alpha) remained unaffected by the addition of IL-8. HIV-1 induction by IL-1beta, IL-6 and TNF-beta, cytokines grouped here as intermediate HIV-1 inducers, was suppressed by IL-8 at concentrations of 1 and 10 ng/ml. However, IL-8 at 100 ng/ml did not significantly alter the effect of IL-1beta, synergized with IL-6 in enhancing, and marginally suppressed TNF-beta-induced HIV-1 expression. IL-8 suppressed granulocyte-macrophage colony-stimulating factor (GM-CSF) and enhanced interferon gamma (IFN-gamma)-induced HIV-1 expression in a dose-dependent manner. Pretreatment of U1 cells with IL-8 did not alter the IL-8-mediated effects on cytokine-induced HIV-1 expression, suggesting that this chemokine exerts its effect at the time of HIV-1 induction or at a postinduction stage. Furthermore, IL-8 was itself induced by cytokines that up-regulate HIV-1 expression in U1 cells and the levels produced correlated directly with the levels of p24 antigen produced, suggesting common pathways for cytokine induction of both HIV-1 and IL 8. These results show that IL-8, typically a non-inducer, can differentially modulate HIV-1 expression in U1 cells and that this is dependent on the inducing cytokine and on the concentration of IL-8. PMID- 11012766 TI - The Rubino test for leprosy is a beta2-glycoprotein 1-dependent antiphospholipid reaction. AB - We describe the isolation and identification of three components required for the Rubino reaction (RR), which is the rapid sedimentation of formalinized sheep red blood cells (SRBC) initiated by serum from leprosy patients with defective Mycobacterium leprae-specific cell immunity. The Rubino reaction factor (RRF) required for this phenomenon, previously identified as an immunoglobulin M (IgM), was purified from leprosy patient serum by adsorption to formalinized SRBC. Purified RRF IgM, when added to formalinized SRBC, did not produce a positive RR. However, when the contact was carried out in the presence of normal human serum (NHS), cells rapidly sedimented. The purified cofactor from NHS contained two components of 70 000 and 50 000 molecular weight (MW), as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The latter was recognized by the RRF IgM on immunoblot and its N-terminal sequence indicated that it was beta2-glycoprotein 1 (beta2-GP1), an anionic phospholipid-binding protein. Methanol-treated formalinized SRBC did not support the RR. Thin-layer chromatography of an extract of membranes indicated that the SRBC ligand was a cell-surface phospholipid. Cardiolipin inhibited the RR. These data demonstrate that the RR involves a trimolecular interaction in which IgM, beta2-GP1 and an SRBC phospholipid participate. By analogy with the antiphospholipid antibodies (anti-PL) that occur in autoimmune processes, serum samples from 29 systemic lupus erythematosus patients with high levels of anticardiolipin antibodies were submitted to the RR. A positive RR was obtained for 45% (13 of 29 patients). These results modify the paradigm of the absolute specificity of the RR for leprosy and demonstrate that RRF IgM is a beta2-GP1-dependent anti-PL. PMID- 11012767 TI - Genetically detoxified mutants of heat-labile enterotoxin from Escherichia coli are effective adjuvants for induction of cytotoxic T-cell responses against HIV-1 gag-p55. AB - There is an urgent need for prophylactic and therapeutic vaccines against human immunodeficiency virus (HIV). Mucosal immunization strategies have great potential to elicit both mucosal and systemic cellular immunity required to protect against HIV-induced acquired immune deficiency syndrome (AIDS). However, mucosal immunizations with soluble protein antigens generally require adjuvants. In this study, we tested two mutants of the heat-labile enterotoxin (LT) from Escherichia coli, LTK63: with no measurable ADP-ribosyltransferase activity, and LTR72: with residual ADP-ribosyltransferase activity, as mucosal adjuvants for induction of cytotoxic T lymphocyte (CTL) responses to coadministered HIV gag p55 protein. We found that intranasal (i.n.) immunizations with HIV gag p55 protein coadministered with LTK63 or LTR72 induced systemic CTL responses comparable to that obtained following intramuscular (i. m.) immunizations with the same adjuvants. Moreover, oral coadministration of LTR72, but not LTK63, resulted in local as well as systemic p55-specific CTL responses in mesenteric lymph nodes (MLN) and spleens (SP) of the immunized mice. These data have important implications for current efforts to develop a safe vaccine against HIV. PMID- 11012768 TI - Intestinal lymphocyte number, migration and antibody secretion in young and old rats. AB - This study demonstrates that the mucosal immune response to cholera toxin (CT) is compromised in old rats in comparison with young animals. The total number of immunoglobulin A (IgA)-secreting cells is similar or higher in the intestinal inductor and effector sites in old animals. However, the number of specifically induced anti-CT IgA antibody-secreting cells is lower in these tissues in comparison with those in young animals. The kinetics of this immune response in the different gut-associated lymphoid tissues studied suggests that the age associated decline in the number of anti-CT IgA-secreting cells in the intestinal mucosa reflects impaired IgA immunoblast migration. Our data from lymphocyte adoptive transfer studies indicate that factors intrinsic to both the donor cells and the host recipient influence the migration of immunoblasts from the Peyer's patches to the effector site. For example, donor cells from old donors transferred to either young or old recipient rats migrate slower than young donor lymphocytes transferred into old host animals. In vitro studies clearly indicate that ageing does not impair antibody secretion by intestinal mucosal plasma cells. Therefore, the age-related decline in the intestinal mucosal immune response, e.g. diminished specific antibody titres in intestinal lavage, reflects fewer antibody-secreting cells in the mucosa. PMID- 11012769 TI - CD28, CTLA-4 and their ligands: who does what and to whom? PMID- 11012770 TI - Phenotypic analysis of peripheral CD4+ CD8+ T cells in the rat. AB - Among peripheral T cells, the expression of CD4 and CD8 is almost mutually exclusive. However, here we show, using flow cytometric analysis, that ex vivo approximately 6% of rat T cells stained for both CD4 and CD8. These double positive cells were also detected by confocal microscopy. Only around 50% of double positive cells expressed the CD8beta chain, the remaining cells expressed the CD8alpha chain alone. Double positive cells were blast-like with a phenotype, distinct from that of either CD4 or CD8 single positive cells, suggestive of an activated state. Previous reports of double positive T cells have also suggested that coexpression of CD4 and CD8 is linked to the activation state of the cell. There was an indication that priming animals with a hapten-carrier complex increased the ratio of CD8alphaalpha : alphabeta expressing double positive T cells, although we did not detect an increase in the frequency of double positive T cells following priming. We also show that the frequency of double positive cells was reduced following thymectomy and with age. In conclusion, these studies show that peripheral T cells expressing both CD4 and CD8 can be detected in the rat and that they are phenotypically distinct from CD4 and CD8 single positive T cells. PMID- 11012771 TI - Staphylococcal enterotoxin B induces potent cytotoxic activity by intraepithelial lymphocytes. AB - In food poisoning, Staphylococcus aureus secretes staphylococcal enterotoxin B (SEB), a superantigen that causes intense T-cell proliferation and cytotoxicity. The effects of SEB on lytic activity by human intestinal intraepithelial lymphocytes (IEL) were investigated. Jejunal IEL, from morbidly obese individuals undergoing gastric bypass operations, were tested for SEB-induced cytotoxicity against C1R B-lymphoblastoid cells, HT-29 adenocarcinoma cells, or CD1d transfected cells using the 51Cr-release assay. Fas and Fas ligand expression were detected by immunofluorescence and flow cytometry and soluble ligand by enzyme-linked immunosorbent assay (ELISA). In the presence of SEB, IEL became potently cytotoxic against C1R cells and interferon-gamma (IFN-gamma)-precultured HT-29 cells, causing 55+/-10% and 31+/-6% lysis, respectively, greater than that by phytohaemagglutinin (PHA)-, interleukin-2 (IL-2)-, or anti-T-cell receptor (TCR)-activated IEL. SEB-stimulated peripheral blood (PB) CD8+ T cells lysed similar numbers of C1R cells but fewer HT-29 cells (53+/-13% and 8+/-5%, respectively). IEL killing of C1R cells involved interaction of major histocompatibility complex (MHC) class II with TCR, CD2 with CD58, and CD11a with CD54, and was perforin mediated. SEB-induced IEL lysis of HT-29 cells, in contrast, was caused by an unknown target cell structure, not MHC class II or CD1d, and resulted from a combination of perforin and Fas-mediated events. The potent cytotoxic activities of IEL promoted by SEB utilize two different mechanisms, depending on the surface receptors expressed by the target cells. PMID- 11012772 TI - HLA-G has a concentration-dependent effect on the generation of an allo-CTL response. AB - Human leucocyte antigen (HLA) -G is expressed on trophoblast cells during pregnancy, suggesting a role in protection of the semiallogeneic fetus. Published data suggest that HLA-G protects a cell against natural killer cell lysis. It has been hypothesized that HLA-G may also protect the fetus by preventing allo cytotoxic T lymphocyte (CTL) responses. To test this hypothesis, we assayed the effects of various concentrations of purified HLA-G on CTL response in a mixed lymphocyte culture (MLC) system. We found that concentrations > or =0.1 microg/ml of HLA-G suppressed the allo-CTL response by 30-100% over the control, but, paradoxically, concentrations of 0.01-0.05 microg/ml of HLA-G augmented the allo CTL response by 25-50% over the control. Concentrations < or = 0.001 microg/ml HLA-G had no effect. Addition of HLA-G to preprimed allo-CTL effector cells did not affect their killing ability. Allo-CTL suppressive doses of HLA-G induced a T helper type 2 (Th2) cytokine response, whereas allo-CTL-enhancing doses of HLA-G induced a Th1-type cytokine response. HLA-G purified from first-trimester placenta does not affect allo-proliferative responses nor does it alter the percentage of CD4+ or CD8+ T cells in MLCs. These findings support a potential role for HLA-G-mediated suppression of allo-CTL formation in normal pregnancies. In addition, the effects observed at lower concentrations of HLA-G may have interesting implications for the condition of pre-eclampsia in which concentrations of this HLA class I molecule are reduced. PMID- 11012773 TI - CD5-positive and CD5-negative human B cells converge to an indistinguishable population on signalling through B-cell receptors and CD40. AB - Whether CD5 on B cells marks a subset functionally distinct from the conventional CD5 negative (CD5neg) adult population or is more an indicator of activation, remains contentious. Here we have investigated whether CD5 positive (CD5pos) and CD5neg B cells can be distinguished in terms of their response to surrogate signals aimed to model, in vitro, T-cell dependent (TD) and T-independent (TI) encounters with antigen in vivo: the predominantly CD5pos B-cell population found in cord blood, CD5 B cells positively selected from tonsils and their CD5neg counterparts, were compared. Neonatal B cells displayed a near-identical phenotype to that of adult CD5pos B cells, being characterized by uniform immunoglobulin M (IgM), immunoglobulin D (IgD), CD23 and CD44 coexpression. When cultured with anti-IgM maintained at high density on CD32-tranfected mouse L cells to model TI responses or on CD40 ligand (CD40L)-bearing L cells (with or without captured anti-IgM) to model TD encounters, DNA synthesis was stimulated to a similar extent in all three populations. Focusing on CD5 and CD23, we found that - although the signals delivered promoted distinct profiles of expression - under each condition of activation, the phenotypes that emerged for adult CD5pos and CD5neg B cells were remarkably similar. Neonatal B cells displayed a greater diminution in CD5 expression than adult CD5pos B cells following CD40 signals but otherwise the two populations again behaved similarly. The inclusion of interleukin-4 (IL-4) to cultures where cells were costimulated via surface (s)IgM and CD40 resulted in a complete loss of CD5 expression and a corresponding hyperexpression of CD23, irrespective of the population studied. The near identical response of CD5pos and CD5neg B cells to surrogate TD or TI signals in vitro and their convergence to indistinguishable phenotypes is wholly supportive of CD5 being a fluctuating marker of activation rather than it delineating functionally distinct subsets. PMID- 11012774 TI - Signals sustaining human immunoglobulin V gene hypermutation in isolated germinal centre B cells. AB - Affinity maturation of antibody responses depends on somatic hypermutation of the immunoglobulin V genes. Hypermutation is initiated specifically in proliferating B cells in lymphoid germinal centres but the signals driving this process remain unknown. This study identifies signals that promote V gene mutation in human germinal centre (GC) B cells in vitro. Single GC B cells were cultured by limiting dilution to allow detection of mutations arising during proliferation in vitro. Cells were first cultured in the presence of CD32L cell transfectants and CD40 antibody (the 'CD40 system') supplemented with combinations of cytokines capable of supporting similar levels of CD40-dependent GC B-cell growth [interleukin (IL)-10 + IL-1beta + IL-2 and IL-10 + IL-7 + IL-4]. Components of the 'EL4 system' were then added to drive differentiation, providing sufficient immunoglobulin mRNA for analysis. Analysis of VH3 genes from cultured cells by reverse transcription-polymerase chain reaction (RT-PCR)-based single-strand conformation polymorphism indicated that the combination IL-10 + IL-1beta + IL-2 promoted active V gene mutation whereas IL-10 + IL-7 + IL-4 was ineffective. This was confirmed by sequencing which also revealed that the de novo generated mutations were located in framework and complementarity-determining regions and shared characteristics with those arising in vivo. Somatic mutation in the target GC B-cell population may therefore be actively cytokine driven and not simply a consequence of continued proliferation. The experimental approach we describe should facilitate further studies of the mechanisms underlying V gene hypermutation. PMID- 11012776 TI - M-ficolin is expressed on monocytes and is a lectin binding to N-acetyl-D glucosamine and mediates monocyte adhesion and phagocytosis of Escherichia coli. AB - Ficolins are a group of multimeric proteins that contain collagen-like and fibrinogen-like (FBG) sequences. Three types of ficolins have been characterized: H-, L- and M-ficolins. Both H- and L-ficolins have demonstrated lectin activities. In the present study, the FBG domain of M-ficolin was expressed and shown to bind to N-acetyl-D-glucosamine. M-ficolin mRNA was expressed in monocytes but not in the more differentiated macrophages and dendritic cells. By flow cytometry, surface biotinylation and immunoprecipitation, we showed that M ficolin was associated with the surface of promonocytic U937 cells. M-ficolin transiently expressed in COS-7 cells was also clearly detected on the cell surface by immunoprecipitation. By flow cytometry, M-ficolin was detected on peripheral blood monocytes but not on lymphocytes or granulocytes. Immobilized rabbit anti-M-ficolin F(ab')2 mediated U937 cell adhesion, and the antibody also inhibited phagocytosis of Escherichia coli K-12 by U937 cells. Therefore, M ficolin might act as a phagocytic receptor or adaptor on circulating monocytes for micro-organism recognition and may potentially mediate monocyte adhesion. PMID- 11012775 TI - Increased immunoglobulin A levels in milk by over-expressing the murine polymeric immunoglobulin receptor gene in the mammary gland epithelial cells of transgenic mice. AB - The polymeric immunoglobulin receptor (pIgR) transports dimeric immunoglobulin A (dIgA) across the epithelial cell layers into the secretions of various mucosal and glandular surfaces of mammals. At these mucosal sites, such as the gastrointestinal tract, respiratory tract, urogenital tract and the mammary glands, dIgA protects the body against pathogens. The pIgR binds dIgA at the basolateral side and transports it via the complex mechanism of transcytosis to the apical side of the epithelial cells lining the mucosa. Here, the extracellular part of the receptor is cleaved to form the secretory component (SC), which remains associated to dIgA, thereby protecting it from degradation in the secretions. One pIgR molecule transports only one dIgA molecule (1 : 1 ratio) and the pIgR is not recycled after each round of transport. This implies that the amount of available receptor could be a rate-limiting factor determining both the rate and amount of IgA transported per cell and therefore determining the total IgA output into the lumen or, in case of the mammary gland, into the milk. In order to test this hypothesis, we set up an in vivo model system. We generated transgenic mice over-expressing the murine pIgR gene under lactogenic control, by using a milk gene promoter, rather than under immunological control. Mice over expressing the pIgR protein, in mammary gland epithelial cells, from 60- up to 270-fold above normal pIgR protein levels showed total IgA levels in the milk to be 1.5-2-fold higher, respectively, compared with the IgA levels in the milk of non-transgenic mice. This indicates that the amount of pIgR produced is indeed a limiting factor in the transport of dIgA into the milk under normal non inflammatory circumstances. PMID- 11012777 TI - Contrasting effects of myeloid dendritic cells transduced with an adenoviral vector encoding interleukin-10 on organ allograft and tumour rejection. AB - Mouse bone marrow-derived myeloid dendritic cells (DC) propagated in granulocyte macrophage colony-stimulating factor and transforming growth factor-beta1 (TGF beta1) (so-called 'TGF-beta DC') are phenotypically immature, and prolong allograft survival. Interleukin-10 (IL-10) has been shown to inhibit the maturation of DC by down-regulating surface major histocompatibility complex (MHC) class II, co-stimulatory and adhesion molecule expression. Genetic engineering of TGF-beta DC to overexpress IL-10 might enhance their tolerogenic potential. In this study, adenoviral (Ad) vectors encoding the mouse IL-10 gene were transduced into B10 (H2b) TGF-beta DC. Transduction with Ad-IL-10 at a multiplicity of infection (MOI) of 50-100 resulted in a modest reduction in the incidence of DC expressing surface MHC class II, CD40, CD80 and CD86. Paradoxically, Ad-IL-10 transduction enhanced the allostimulatory activity of DC in mixed leucocyte reactions and cytotoxic T lymphocyte assays, and increased their natural killer cell stimulatory activity. Systemic injection of normal C3H recipients with Ad-IL-10-transduced B10-DC 7 days before organ transplantation, exacerbated heart graft rejection and augmented circulating anti-donor alloantibody titres. Contrasting effects were observed in relation to tumour growth. All mice preimmunized with Ad-IL-10-transduced, tumour antigen (B16F10) pulsed DC developed palpable tumours, associated with significant inhibition of splenic anti-tumour cytotoxic T lymphocyte generation. Animals pretreated with control Ad-LacZ-transduced, B16F10-pulsed DC however, remained tumour free. These findings are consistent with the multifunctional immunomodulatory properties of mammalian IL-10. PMID- 11012778 TI - Major histocompatibility complex class II- fetal skin dendritic cells are potent accessory cells of polyclonal T-cell responses. AB - Whereas dendritic cells (DC) and Langerhans cells (LC) isolated from organs of adult individuals express surface major histocompatibility complex (MHC) class II antigens, DC lines generated from fetal murine skin, while capable of activating naive, allogeneic CD8+ T cells in a MHC class I-restricted fashion, do not exhibit anti-MHC class II surface reactivity and fail to stimulate the proliferation of naive, allogeneic CD4+ T cells. To test whether the CD45+ MHC class I+ CD80+ DC line 80/1 expresses incompetent, or fails to transcribe, MHC class II molecules, we performed biochemical and molecular studies using Western blot and polymerase chain reaction analysis. We found that 80/1 DC express MHC class II molecules neither at the protein nor at the transcriptional level. Ultrastructural examination of these cells revealed the presence of a LC-like morphology with indented nuclei, active cytoplasm, intermediate filaments and dendritic processes. In contrast to adult LC, no LC-specific cytoplasmic organelles (Birbeck granules) were present. Functionally, 80/1 DC in the presence, but not in the absence, of concanavalin A and anti-T-cell receptor monoclonal antibodies stimulated a vigorous proliferative response of naive CD4+ and CD8+ T cells. Furthermore, we found that the anti-CD3-induced stimulation of naive CD4+ and CD8+ T cells was critically dependent on the expression of FcgammaR on 80/1 DC and that the requirement for co-stimulation depends on the intensity of T-cell receptor signalling. PMID- 11012779 TI - Granulocyte-macrophage colony-stimulating factor regulates cytokine production in cultured macrophages through CD14-dependent and -independent mechanisms. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has multiple effects on the antigen phenotype and function of macrophages. In this study we investigated the effect of GM-CSF on cytokine production by macrophages. We found that GM-CSF may modify the tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) response to lipopolysaccharide (LPS) through two different mechanisms. Relatively early in culture, GM-CSF increases the amount of cytokines synthesized by responding cells; this effect appears to be unrelated to modulation of CD14 expression and LPS-binding capacity. After prolonged incubation, GM-CSF up regulates both CD14 expression and LPS-binding capacity, and the frequency of cytokine-producing cells. Release of CD14 in the culture supernatant was decreased in the presence of GM-CSF, suggesting that a reduced shedding was responsible for the effect of GM-CSF on CD14 expression. Enhancement of cytokine production was also observed in GM-CSF-treated macrophages after stimulation by phorbol 12-myristate 13-acetate (PMA), thus indicating that GM-CSF affects both CD14-dependent and -independent cytokine production. Finally, GM-CSF did not modulate the LPS- and PMA-induced production of IL-10 and IL-12. We conclude that GM-CSF may play a role in manipulating the activation-induced expression of pro inflammatory cytokines by macrophages. Enhanced production of these cytokines could play an important role in the pathogenesis of Gram-negative septic shock syndrome and in defence against infectious agents. PMID- 11012780 TI - The lipid A region of lipopolysaccharides from Rhizobiaceae activates bone marrow granulocytes from lipopolysaccharide-hyporesponsive C3H/HeJ and C57BL/10ScCr mice. AB - We established in previous studies that the binding of Salmonella lipopolysaccharide (LPS) to constitutive receptors of low affinity triggers the expression of the inducible LPS-binding molecule CD14 in bone marrow cells (BMC) of C3H/HeOU mice, but not in BMC from C3H/HeJ mice. We show in this study that BMC from C3H/HeJ and C57BL/10ScCr mice do not express CD14 after exposure to LPSs from Salmonella enterica and Bordetella pertussis, but do express this marker when treated with several LPSs from Rhizobiaceae, or their lipid A fragments. This shows that the constitutive LPS receptor in BMC from C3H/HeJ and C57BL/10ScCr mice is fully able to trigger a complete signalling cascade. Results of cross-inhibition of the binding of radiolabelled LPS indicated that active LPSs (from R. species Sin-1 and R. galegae) and inactive LPSs (from S. enterica and B. pertussis) bind to the same site of the constitutive LPS receptor of C3H/HeJ cells. Furthermore, binding of R. species Sin-1 LPS, and signalling induced by this LPS, were both inhibited by pre-exposure of C3H/HeJ cells to B. pertussis lipid A. This correlation between binding and signalling suggests that in C3H/HeJ cells, the constitutive receptor, which recognizes a large panel of LPSs from different origins, appears selectively unable to be activated by some particular LPSs, such as those of Enterobacteria and Bordetella. PMID- 11012781 TI - Human endothelial cells are activated by interferon-gamma plus tumour necrosis factor-alpha to kill intracellular Pseudomonas aeruginosa. AB - Proinflammatory cytokines have been shown to activate endothelial cells. To investigate the effect of cytokines on the interaction of human umbilical vein endothelial cells (HUVEC) with Pseudomonas aeruginosa, cells were treated with interferon-gamma (IFN-gamma) plus tumour necrosis factor-alpha (TNF-alpha) for 24 hr and exposed to P. aeruginosa suspension for 1 hr. Light microscopy showed that activated cells internalized significantly more bacteria than control cells. To ascertain the effect of cytokines on the microbicidal activity of HUVEC, the concentrations of viable intracellular (IC) bacteria in control and activated cells were determined, at 1 and 5 hr postinfection, by the gentamicin exclusion assay. In control cells, no significant decrease in the concentration of bacteria was detected 5 hr postinfection. In contrast, in activated cells the concentration of viable bacteria at 5 hr was significantly lower. Concentrations of superoxide and hydrogen peroxide detected in supernatants of activated cells were significantly higher than in control cell supernatants. HUVEC anti-P. aeruginosa activity was insensitive to the antioxidants superoxide dismutase, dimethylthiourea and allopurinol as well as to the L-arginine analogues aminoguanidine and NG-monomethyl-L-arginine (L-NMMA), but was significantly inhibited by catalase. Our results indicate that HUVEC can be activated by IFN gamma plus TNF-alpha to kill IC P. aeruginosa and suggest a role for reactive oxygen radicals, notably hydrogen peroxide, in HUVEC antibacterial activity. PMID- 11012782 TI - Soluble isoforms of CEACAM1 containing the A2 domain: increased serum levels in patients with obstructive jaundice and differences in 3-fucosyl-N-acetyl lactosamine moiety. AB - CEACAM1 (biliary glycoprotein or CD66a) is a member of the carcinoembryonic antigen (CEA) subgroup of the CEA family. Eleven RNA isoforms derived from the splicing of a single CEACAM1 gene have been described. Some of the CEACAM1 isoforms have been recognized by the CD66 antibodies in T and B lymphocytes, natural killer cells, granulocytes and epithelial cells in several human tissues. Although it is also present in soluble form in bile and serum, and elevated levels have been found in the serum of patients with liver diseases, it is not known which isoforms are primarily involved. In order to learn more about the distribution and properties of particular CEACAM1 isoforms, we have prepared a monoclonal antibody specific for the A2 domain of CEACAM1, designated TEC-11. This antibody does not cross-react with other members of the CEA family. Immunoblotting analysis revealed that the TEC-11 epitope was present in all cell types expressing CEACAM1 containing the A2 domain [CEACAM1(A2)], including granulocytes (160 000 MW isoform) and sperm cells (140 000 MW isoform). A 115 000 MW isoform of CEACAM1(A2) was present in human serum, bile, saliva and seminal fluid. Human bile, saliva and seminal fluid also contained the 160 000 MW CEACAM1(A2) isoform. Significantly higher serum levels of the 115 000 MW CEACAM1(A2) isoform were detected in patients with obstructive jaundice. The 160 000 MW isoform of CEACAM1(A2) in bile, but not a 115 000 MW isoform in serum and bile, carried the 3-fucosyl-N-acetyl-lactosamine moiety. The combined data indicate that various isoforms of CEACAM1(A2) are present in different body fluids where they could take part in different CEACAM1-mediated functions. PMID- 11012783 TI - Specificity of a new lipid mediator produced by testicular and peritoneal macrophages on steroidogenesis. AB - Macrophage-derived factor (MDF) is a lipophilic factor produced by rat testicular and peritoneal macrophages that maximally stimulates testosterone production by rat Leydig cells through a steroidogenic acute regulatory protein independent mechanism. The purpose of the present study was to determine whether MDF is also produced by human macrophages, and/or if it acts on human steroidogenic cells. We also studied the tissue-specific functions of MDF by determining if it also acts on steroidogenic cells of the ovary and adrenal glands and, if so, does it require new protein synthesis. It was found that MDF was produced by human peritoneal macrophages, and was capable of stimulating human steroidogenic cells. In terms of tissue specificity, it was found that primary cultures of rat adrenocortical cells respond to MDF with increased secretion of aldosterone and corticosterone, as did rat granulosa cells by producing progesterone. MDF acted in the presence of cycloheximide, indicating that it does not require new protein synthesis. These results indicate that MDF may have significant therapeutic potential and provide a basis for future studies concerning its physiological role in humans. These results further suggest that MDF is not only involved in paracrine regulation of Leydig cells, but also has the potential for the local regulation of steroidogenesis in both granulosa and adrenal cortical cells. PMID- 11012784 TI - Marmoset spermatogenesis: organizational similarities to the human. AB - The common marmoset (Callithrix jacchus), a small New World primate of high fecundity, is widely used in reproductive research. The aim of the present study was to determine the organization of the germ cells within the seminiferous epithelium, the duration of the spermatogenic cycle and the number of spermatogonial mitoses. Antibodies to cAMP response element modulator (CREM) and proliferating nuclear cell antigen (PCNA) and a cRNA directed against protamine P2 and morphological criteria were used to discriminate between stages of the spermatogenic cycle. Plastic sections were used to document the cell associations present in each of the nine stages of spermatogenesis. Up to five such stages could be observed within individual cross-sections of seminiferous tubules. Based on the pattern of incorporation of bromodeoxyuridine the length of the spermatogenic cycle was estimated to be 10 days and the duration of spermatogenesis to be 37 days. Four mitotic divisions were noted in spermatogonia. It is concluded that the organization of spermatogenesis in the marmoset has similarities to the human ('helical') and this makes the marmoset a suitable model for studies relevant to human testicular function. PMID- 11012785 TI - Immunohistochemical localization of V-ATPases in rat spermatids. AB - The sperm acrosome exhibits a low pH. However, the mechanism of acidification in the acrosome remains unclear. Vacuolar-type proton ATPase (V-ATPase) has been shown to play a principle role in generating and maintaining the acidity of organelles such as lysosomes and endosomes. In this study, we examined whether V ATPase is localized in the acrosome membranes using immunohistochemical techniques. Sections of rat testis were immunostained using antibodies against V ATPase. Under light microscopic observation, the perinuclear region in spermatids at an early stage of development was heavily immunostained. At the electron microscopic level, gold particles showing the presence of V-ATPase were localized to the acrosome membranes in the developing spermatids. V-ATPase was also localized to the membrane of vesicles locating between the trans-Golgi area and the acrosome. These observations suggest that V-ATPase may play a role in acrosome acidification. PMID- 11012786 TI - In vitro inhibition of rat cauda epididymal sperm glycolytic enzymes by ornidazole, alpha-chlorohydrin and 1-chloro-3-hydroxypropanone. AB - Chlorinated antifertility compounds are known to inhibit glycolysis of spermatozoa as they reside in the epididymis but new compounds need to be evaluated that retain antifertility action but do not exhibit side-effects. In this study, two known antifertility agents and a related compound were compared for their inhibition of rat sperm metabolism and motility in vitro. The dose dependent inhibition in vitro of the glycolytic enzymes glyceraldehyde-3 phosphate dehydrogenase (GAPDH) and triosephosphate isomerase (TPI) of distal cauda epididymal rat spermatozoa by (R)-, (S)- and (R,S)-ornidazole (ORN), (R,S) alpha-chlorohydrin (ACH) and 1-chloro-3-hydroxypropanone (CHOP) was compared. The direct inhibition of GAPDH by ORN suggests that it inhibits without prior conversion outside the cell but inhibition was not stereo-specific. The GAPDH, but not TPI, activity of spermatozoa incubated with ACH and CHOP was highly correlated with kinematic parameters of spermatozoa incubated in pyruvate- and lactate-free medium. ACH only inhibited the activity of intact spermatozoa and the inhibition was not reversed by washing the particulate sperm fraction after sonication. High concentrations of ACH (100 mmol/L) killed intact rat spermatozoa and decreased the extent of GAPDH inhibition. CHOP, unlike ACH, was an effective inhibitor of both intact and sonicated cells. Pre-CHOP, the dimethylketal precursor of CHOP, and its other hydrolysis product MeOH, were both ineffective in vitro. CHOP and related ketals may be more effective inhibitors of sperm glycolysis than ACH and may prove useful for investigating sperm-specific glycolytic inhibition, a prerequisite for the development of antiglycolytic, post testicular acting contraceptives. PMID- 11012787 TI - Immunocytochemical detection of mitochondrial dihydroorotate dehydrogenase in human spermatozoa. AB - In mammalian cells the requirement for pyrimidines is met by uridine phosphate (UMP) de novo synthesis and, to a greater or lesser extent, by salvage of free nucleosides. The fourth enzyme of the de novo synthesis, the mitochondrially bound dihydroorotate dehydrogenase (DHODH) was the focus of the present study. Rabbit anti-DHODH IgG, which was generated using an immunization protocol with truncated recombinant human DHODH protein and purified by an immunosorbent method, was used for immunocytochemical detection and localization of this enzyme in ejaculated human spermatozoa. The presence of DHODH protein was demonstrated by Western blotting of solubilized membrane fractions with peroxidase conjugated anti-rabbit IgG in combination with chemiluminescence detection. Indirect immunofluorescence microscopy, using Cy3-conjugated anti-rabbit IgG, revealed specific binding in the midpiece of spermatozoa. As these cells no longer have a demand for de novo biosynthesis of pyrimidines, we hypothesize that the pathway could serve a specialized function in nitrogen or zinc metabolism during the process of spermiogenesis and/or epididymal maturation. PMID- 11012788 TI - Comparative study of disomy and diploidy rates in spermatozoa of fertile and infertile men: a donor-adapted protocol for multi-colour fluorescence in situ hybridization (FISH). AB - Spermatozoa from seven healthy donors (two of whom had already fathered children) and five infertile patients taking part in the local programme of intracytoplasmic sperm-injection (ICSI) were investigated for the disomy rates of chromosomes 13/21, 18, X and Y as well as for the diploidy rates. Two- and three colour fluorescence in situ hybridization (FISH) was applied after a donor adapted decondensation pre-treatment: in a preliminary decondensation series the optimum fluorescence signals were individually determined by variation of the concentration of the decondensation reagents and the duration of incubation with these reagents. Strict scoring criteria were applied. The average disomy rates ranged from 0.10% (chromosomes 13/21) to 0.44% (disomy XY) in the infertile donors and from 0.07% (disomy XX) to 0.36% (disomy XY) in the controls. The average diploidy rates were 0.22% and 0.20% for the infertile donors and the controls respectively. There was no statistically significant difference between the two groups with respect to the disomy and diploidy rates. Within the two groups there were inter-individual differences which were partly statistically significant, indicating considerable inter-donor variation of the aneuploidy rates. PMID- 11012789 TI - Diurnal scrotal skin temperature and semen quality. The Danish First Pregnancy Planner Study Team. AB - It is well established that heat is associated with reduced sperm production, but the role of physiological variation in temperature has never been scrutinized in humans. We studied diurnal scrotal temperature and markers of male fertility in a population of couples planning their first pregnancy. Sixty men from a cohort of couples who were planning their first pregnancy were included and scrotal skin temperature was monitored during 3 days using a portable data recorder. Working hours and working postures were recorded daily in a questionnaire. Each man provided a fresh semen sample and the couples were followed for six menstrual cycles or until a clinical pregnancy was recognized. The median value of scrotal skin temperature was 33.3 degrees C in the daytime and 34.8 degrees C at night. In periods of sedentary work, the median temperature was on average 0.7 degrees C higher (SE=0.2 degrees C). In addition, scrotal temperature was higher in the daytime, in summer, and in leisure time compared with working hours. Median sperm concentration among men with more than 75% of their daytime readings above 35 degrees C was 33.4 x 10(6)/mL, compared with 91.8 x 10(6)/mL for men with less than half of their readings above 35 degrees C (difference 58.4; 95% CI: 25.9 77.8 x 10(6)/mL). It is concluded that a sedentary position is a significant source of increased scrotal skin temperature, and even moderate and physiological elevation in scrotal skin temperature is associated with a substantially reduced sperm concentration. Sedentary work should be considered as an important potential confounder for reduced sperm count in epidemiological research. PMID- 11012791 TI - This month in JAN PMID- 11012790 TI - Addiction: global problem and global response. Complacency or commitment. PMID- 11012792 TI - Cognitive therapy supervision as a framework for clinical supervision in nursing: using structure to guide discovery. AB - Cognitive therapy supervision as a framework for clinical supervision in nursing: using structure to guide discovery Cognitive therapy has an undisputed evidence base upon which its clinical application flourishes. This approach is now a well recognized and widely adopted method used in the treatment of a diversity of psychological problems. More recently, prominent innovators of this psychotherapy have devised a framework to guide the clinical supervision of cognitive therapists. In keeping with its therapeutic application, the cognitive therapy framework for supervision is focused, structured, educational and collaborative. It serves to enhance the therapeutic proficiency of the cognitive therapist. In contrast, the supervision models reported in the recent nursing literature are less precise in their mission and when evaluated their contribution to nursing is shown to be dubious. Following an overview of the supervision models commonly cited in the nursing literature, a more focused comment on the evaluative research concerning Proctor's three-function interactive model will be offered. It is suggested that the unconvincing research findings may be related to the conceptual muddle surrounding clinical supervision, and the expectation for clinical supervision to deliver more than an opportunity for the progression of our therapeutic integrity. From this, a cognitive therapy supervision framework is described and suggested by the authors as a structure from which supervisors can guide discovery. PMID- 11012793 TI - An evaluation of nurse rostering practices in the National Health Service. AB - An evaluation of nurse rostering practices in the National Health Service The scheduling of nursing time on hospital wards is critical to the delivery of patient care, resource utilization and employee satisfaction. Over the past decade many hospital wards in the United Kingdom (UK) have moved away from the traditional planning of rosters by a single manager, towards more participative processes known as self-rostering and team rostering. This paper tests the hypothesis, developed from the literature, that the three types of rostering approach may be positioned along a continuum. Self-rostering at one extreme, is conducive to staff empowerment, motivation and roster effectiveness, whilst departmental rostering, at the other, leads to perceived autocracy, reduced empowerment, lower levels of staff motivation and roster effectiveness. Team rostering is positioned mid-way on this continuum. This paper reports the findings of an empirical study of nurse rostering practices in the UK National Health Service (NHS), with a view to developing an understanding of the implications of implementing these three rostering approaches and testing the above hypothesis. The survey of rostering practices in 50 NHS wards, and in-depth case studies of seven wards, revealed that each of the three rostering approaches has benefits and limitations and a picture emerges quite different from that implied by the research hypothesis. Whilst the literature suggests that the choice of rostering approach determines the level of perceived autocracy, staff motivation and roster effectiveness, it is proposed in this paper that selection of rostering approach should be contingent upon operational context. The paper concludes with a framework which stipulates that the choice of rostering approach for a ward should be determined on the basis of four contingent variables, namely, ward size, demand variability, demand predictability, and complexity of skill mix. It is recommended that departmental rostering be applied in large wards with complex rostering problems, whilst team rostering is more appropriate for medium sized wards, and self-rostering appropriate for small wards. PMID- 11012794 TI - Hospital nurses' job satisfaction, individual and organizational characteristics. AB - Hospital nurses' job satisfaction, individual and organizational characteristics Using the Ward Organizational Features Scales (WOFS), relationships between aspects of the organization of acute hospital wards, nurses' personal characteristics and nurses' job satisfaction are examined among a nationally representative sample of 834 nurses in England. The analysis contributes to a growing body of evidence demonstrating the importance of interpersonal relationships to nurses' job satisfaction. In particular, the positive contribution of the cohesiveness of ward nursing staff is highlighted, but the potential for many current NHS staffing strategies and work environments to undermine the development of cohesive working relationships is also noted. Other influential factors are nurses' relationships with medical staff, perceptions of their workload and their evaluation of the appropriateness of the system of nursing being practised. The importance of measuring nurses' subjective assessments of their work environment is emphasized. A weak association was found between grade and job satisfaction. Individual nurse characteristics were found not to be associated with job satisfaction. PMID- 11012795 TI - The current scope and future direction of perioperative nursing practice in Victoria, Australia. AB - The current scope and future direction of perioperative nursing practice in Victoria, Australia Unlike some overseas countries, perioperative nursing roles in Australia are largely undeveloped, and very little research has explored the opinions of perioperative nurses regarding the development of their specialty. This study was undertaken to identify the current scope, possible future directions, and the barriers to developing perioperative nursing practice in the state of Victoria (Australia). The study design was descriptive and utilized survey methodology. An instrument was constructed by the researcher for the specific purposes of the study, and comprised three stages of development: questionnaire development; validation by a panel of experts; and a pilot study. The questionnaire was sent randomly to selected perioperative nurses (n=224) and a response rate of 79% (n=176) was obtained. Data were analysed using the Statistical Package for the Social Sciences (SPSS). Descriptive statistics were used to summarize data, and independent sample t-tests, one-way ANOVA and Wilcoxon signed rank tests were used to test differences between groups. For the purpose of data reduction factor analysis was undertaken. The findings of the study indicated that the perioperative nursing model in its espoused form is not being practised, but a large majority of participants believe that perioperative nursing practice should not be restricted to the intraoperative phase of the perioperative model. Expanded rather than extended activities and functions were perceived as more important for development of the specialty, and perceived barriers to advanced perioperative nursing practice centred predominantly on the current application of the perioperative model. Recommendations include refinement of the perioperative nursing model to reflect changes in the delivery of healthcare, addressing issues of cost associated with further education, and undertaking interpretive research to provide more detailed information on the possible future direction for perioperative nursing. PMID- 11012796 TI - A comparative study of the financing, provision and quality of care in nursing homes. The approach of four European countries: Belgium, Denmark, Germany and the Netherlands. AB - A comparative study of the financing, provision and quality of care in nursing homes. The approach of four European countries: Belgium, Denmark, Germany and the Netherlands As result of an increase in the numbers of frail elderly people, most European countries are facing problems with the financing and provision of services by nursing homes. At the same time, the expectations of quality of these services continue to rise. The main question investigated in this study was that of how countries approach the problems of financing and service provision by nursing homes and, at the same time, attempt to increase the quality levels in these institutions. The study was conducted in Belgium, Denmark, Germany and the Netherlands. A study was made of the relevant literature and questionnaires were sent to experts in each country. The four countries are addressing the problems of financing and sufficient service provision by controlling the use of nursing home services. In addition, financial problems are approached by extending co payments, encouraging cheaper forms of care and putting pressure on nursing homes to operate at lower costs. Problems in the provision of care are addressed by applying more selective admission criteria and offering alternative forms of care outside the nursing home. As a result nursing home beds are used for those with the greatest care-dependency. Nursing home services are adjusted to rising quality expectations by offering a greater range of provision, decreasing the number of residents per room, improving comfort and improving the training of nursing staff. Another way to increase the quality of care is to separate the housing and service functions. Many nursing homes nowadays collaborate intensively with other facilities for the elderly to cope with all these problems and changes. The workload for nursing home staff has increased because of the increasing care-dependency of residents, the demand for higher quality of services and the financial problems. PMID- 11012797 TI - Quality in health care and ethical principles. AB - Quality in health care and ethical principles The last three decades have seen rapid changes in the way United States of America (USA) health care has been delivered, financed and regulated. Four major stakeholders have emerged in the health care debate: patients, providers, payers and public regulatory agencies. These groups do not agree on a definition of quality health care. This paper suggests five ethical principles - autonomy, justice, beneficence, non maleficence, and prudence - be included in the framework of quality health care. A framework that outlines possible relationships among these ethical attributes and four major stakeholders is presented. PMID- 11012798 TI - Protecting children: intuition and awareness in the work of health visitors. AB - Protecting children: intuition and awareness in the work of health visitors This paper is based upon an ethnographic study of a group of health visitors engaged in child protection work. The purpose of this paper is to explore the meanings individual health visitors attach to events concerned with identifying children who may be at risk of harm from child abuse, and also the idiosyncratic nature of health visiting in this complex but everyday social situation. The paper focuses on understanding the importance of a particular form of knowledge which the health visitors referred to as 'intuitive awareness'. PMID- 11012799 TI - Who cares? Offering emotion work as a 'gift' in the nursing labour process. AB - Who cares? Offering emotion work as a 'gift' in the nursing labour process The emotional elements of the nursing labour process are being recognized increasingly. Many commentators stress that nurses' 'emotional labour' is hard and productive work and should be valued in the same way as physical or technical labour. However, the term 'emotional labour' fails to conceptualize the many occasions when nurses not only work hard on their emotions in order to present the detached face of a professional carer, but also to offer authentic caring behaviour to patients in their care. Using qualitative data collected from a group of gynaecology nurses in an English National Health Service (NHS) Trust hospital, this paper argues that nursing work is emotionally complex and may be better understood by utilizing a combination of Hochschild's concepts: emotion work as a 'gift' in addition to 'emotional labour'. The gynaecology nurses in this study describe their work as 'emotionful' and therefore it could be said that this particular group of nurses represent a distinct example. Nevertheless, though it is impossible to generalize from limited data, the research presented in this paper does highlight the emotional complexity of the nursing labour process, expands the current conceptual analysis, and offers a path for future research. The examination further emphasizes the need to understand and value the motivations behind nurses' emotion work and their wish to maintain caring as a central value in professional nursing. PMID- 11012800 TI - The contexts of adherence for African Americans with high blood pressure. AB - The contexts of adherence for African Americans with high blood pressure African American men between the ages of 18 and 49 years have the lowest rates of awareness, treatment and control of high blood pressure (HBP) of all age/race/gender groups in the United States. A qualitative study was done to gain an understanding of urban black males' experiences of living with HBP. In-depth semi-structured interviews were conducted with 19 black males. The interviews explored perceptions of health, health problems and priorities, and concerns of daily living that influenced appointment keeping and medication taking. The sample was a subset of 309 men participating in a 3-year clinical trial to improve HBP control in an inner city African-American population. Content analysis of transcribed interviews identified the following themes and related concerns: (a) personal contexts: meaning of health, high blood pressure and treatments; (b) social context: living as a young black male in an urban environment; and (c) cultural context of relating: patient-provider relationship can make a difference. Influencing participants' responses were: interpreting symptoms; adjusting medication taking; protecting personal privacy; allocating limited resources; dealing with addiction; and feeling cared for by a health care provider. Adherence appeared to be multifaceted and changing depending upon: the men's social, economic and personal circumstances; empathetic and non-judgemental assistance from providers; financial concerns and employment; and drug addiction. Findings are useful in refining high blood pressure interventions. PMID- 11012801 TI - Re-thinking stroke rehabilitation: the Corbin and Strauss chronic illness trajectory framework. AB - Re-thinking stroke rehabilitation: the Corbin and Strauss chronic illness trajectory framework The dramatic effects of a stroke can have far-reaching implications for patients and carers. Effective recovery involves a considerable array of coping strategies that facilitate and promote engagement in the social world. Their development is a long-term process that requires considerable effort, motivation and enterprise on the part of patients and their families. Traditional approaches to the provision of stroke rehabilitation services, however, appear to be underpinned by frameworks that are short-term in outlook. As a consequence, nursing interventions often focus on the progression of the patient through the care system, rather than on facilitating future recovery. Much of the work of stroke recovery is consequently done by patients and their families at home, with little provision of ongoing professional help and advice. This paper explores the application of the Corbin and Strauss Chronic Illness Trajectory Framework for stroke. In particular, the major concepts of the framework are applied to a vignette derived from a longitudinal study of patients' experiences of recovery. The trajectory framework is shown to be a useful structure that has the potential to enhance the appropriateness of nursing interventions for stroke patients. However, the validity of the framework can only be established through its application and evaluation in clinical practice. The purpose of this paper is to contribute to a debate that encourages consideration of the framework's utility for nurses to enhance the stroke rehabilitation experience. PMID- 11012802 TI - Pattern recognition as a caring partnership in families with cancer. AB - Pattern recognition as a caring partnership in families with cancer The purpose of this study was to address the process of a caring partnership by elaborating pattern recognition as nursing intervention with families with cancer. It is based on Newman's theory of health as expanding consciousness within the unitary transformative paradigm and is an extension of a previous study of Japanese women with ovarian cancer. A hermeneutic, dialectic method was used to engage 10 Japanese families in which the wife-mothers were hospitalized because of cancer diagnosis. The family included at least the woman with cancer and her primary caregiver. Each of four nurse-researchers entered into partnership with a different family and conducted three interviews with each family. The participants were asked to describe the meaningful persons and events in their family history. The family's story was transmuted into a diagram of sequential patterns of interactional configurations and shared with the family at the second meeting. Evidence of pattern recognition and insight into the meaning of the family pattern were identified further in the remaining meetings. The data revealed five dimensions of a transformative process. Most families found meaning in their patterns and made a shift from separated individuals within the family to trustful caring relationships. One-third of them went through this process within two interviews. The families showed increasing openness, connectedness and trustfulness in caring relationships. In partnership with the family, each nurse researcher grasped the pattern of the family as a whole and experienced the meaning of caring. Pattern recognition as nursing intervention was a meaning making transforming process in the family-nurse partnership. PMID- 11012803 TI - Nurses' experiences of being present with a patient receiving a diagnosis of cancer. AB - Nurses' experiences of being present with a patient receiving a diagnosis of cancer This paper reports the findings of a study which describes the experiences of nurses who were present with a patient when they received a diagnosis of cancer, cancer recurrence or prognosis of terminal cancer. Semi-structured interviews were conducted with six nurses who had experience of caring for patients with cancer in an acute surgical setting. Data were analysed using a phenomenological descriptive approach. Participants' descriptions revealed the following seven core themes: 'What if it was me?'; divergent feelings; being there; becoming closer; method of disclosure; time as an influence and learning by reflection. Possible implications for nursing practice and education are discussed and recommendations are made for future research. PMID- 11012804 TI - The needs of family caregivers of frail elders during the transition from hospital to home: a Taiwanese sample. AB - The needs of family caregivers of frail elders during the transition from hospital to home: a Taiwanese sample This study explored the needs of family caregivers during the transition from hospital to home. Data from 37 face-to-face interviews with 16 caregivers before discharge and at 2 weeks and 1 month after discharge were subjected to constant comparative analysis. Findings revealed changes in family caregivers' needs during the discharge transition. While preparing to take caregiving responsibility, caregivers reported a need for various types of information. After the discharge, but before home caregiving settled into a pattern, they needed help with caregiving practices. After reaching a pattern for providing family care, caregivers frequently reported needs for continuous emotional support. These findings provide a basis for transitional care services such as discharge planning and home health care services. PMID- 11012805 TI - Family and staff perceptions of the role of families in nursing homes. AB - Family and staff perceptions of the role of families in nursing homes Admission to a nursing home is generally regarded as the termination of family care and the commencement of institutional care. Research suggests that following placement families are often expected to relinquish their dependent older relative to the bureaucracy of the institution. The aim of this study was to investigate family and nursing home staff perceptions of the role of families caring for residents in nursing homes. A convenience sample of 44 family carers and 78 nursing home staff completed questionnaires, and interviews were conducted with 10 family carers and 10 nursing home staff. The results suggest that family carers perceived themselves to have a greater role in caring for relatives than that perceived by the nursing home staff. Either families overestimated their involvement, or staff underestimated family involvement in caring for residents in nursing homes. Families were mostly satisfied with their role and with the care provided in nursing homes. They perceived nurses as providers of technical care and they perceived themselves as having an important role in providing social and emotional care. Families trusted the clinical judgement of the staff but the staff were reluctant to trust family carers, especially in situations where care involved an element of risk. Family roles were limited by members' own ability to care and the dependency of the resident, while professional responsibility and accountability discouraged nurses from sharing some caring roles. The results indicate that families in this study were more willing to help in nursing home care and were perhaps under-valued as a resource within the nursing home setting. PMID- 11012806 TI - Psychiatric care as seen by the attempted suicide patient. AB - Psychiatric care as seen by the attempted suicide patient This study highlights the experiences of patients during in-patient psychiatric care in Sweden following a suicide attempt. Eighteen patients were interviewed as close to being discharged as possible. Each respondent was asked to narrate his/her experiences of the care received. An interview guide concerned the following areas: admission to the hospital, feelings and reactions, and positive as well as negative experiences during the hospital stay. The interviews were transcribed verbatim and a qualitative content analysis concerning the meanings, intuitions, consequences and the context of the data was performed. Three central categories were identified: being a psychiatric patient, patients' perceptions of the caregivers and the care provided, as well as important aspects of the psychiatric care received. The importance of being well cared for and receiving understanding and confirmation was emphasized. Lack of confirmation may have contributed in some cases to a feeling of being burdensome, demands for discharge or even another suicide attempt. Verbal contacts with the staff were seen as essential for the process of healing and for the desire to go on living. PMID- 11012807 TI - Determinants of absconding by patients on acute psychiatric wards. AB - Determinants of absconding by patients on acute psychiatric wards Absconding by patients from acute psychiatric wards is a high risk behaviour and has been linked to harm to self and others. Previous research on the characteristics of absconders has been overly reliant on officially generated statistics and small numbers of variables, limiting the conclusions that may be drawn. This paper reports on a prospective study of absconders from 12 acute admission wards in three English National Health Service Trusts over 5 months, compared to a control group matched for ward. Extensive data on absconder and control characteristics were collected from case records and from nursing staff. Absconders were significantly different from controls in many respects. Absconding is linked to other forms of non-compliant patient behaviour, e.g. medication refusal and involvement in violent incidents. Significant variations in the rates of absconding were found between different wards, and between different consultant psychiatrists. Predictive factors were identified by logistic regression. Study in the diverse fields of non-compliance should be brought together as these phenomena are likely to be interrelated. Further investigation is required to determine exactly what it is that consultant psychiatrists and ward nurses do that affects absconding rates. PMID- 11012808 TI - Woodlands therapy: an ethnographic analysis of a small-group therapeutic activity for people with moderate or severe dementia. AB - Woodlands therapy: an ethnographic analysis of a small-group therapeutic activity for people with moderate or severe dementia This paper reports on an analysis of Woodlands therapy (WT), a sensory-motor therapeutic activity that aims to encourage interaction with people with moderate or severe dementia. The aims of the study were to examine: the responses of patients to the sensory experiences and play-based activities that make up a WT session; the factors that influenced their responses; and the verbal interventions employed by the staff who facilitated WT sessions. Methods derived from ethnography were used. The data comprised videotapes of five WT sessions, notes of other sessions, and discussions with staff involved with the approach. Thematic groups of patients' responses were formulated. WT mainly appears to engender the responses of attention, participation and comment. Factors that influence patients' responses are discussed. The findings indicate that staff facilitation strategies strongly influence patients' responses to WT. Verbal interventions made by staff are grouped in terms of their relative helpfulness for promoting positive responses to WT. Suggestions for practice are made for professional carers undertaking sensory-motor therapeutic activities for people with dementia. PMID- 11012809 TI - Correlates of safer sex communication among college students. AB - Correlates of safer sex communication among college students The purpose of this study was to examine factors that are thought to promote communication about safer sex and HIV among college students in the United States of America and to determine the extent to which communication about safer sex is important in the use of condoms. A better understanding of factors associated with safer sex communication can be helpful in developing HIV and STD prevention programmes for college students. Following approval from the institutional review boards of the six participating colleges and universities, researchers collected data from a random sample of students. The study included participant responses if participants were 18-25 years of age, single and sexually active. For the sample of 1349 participants, the mean age was 20.6 years. Sixty-three per cent of the sample was female, 50.5% white, 42.3% African-American, and the remainder of other ethnic groups. Over 50% of respondents reported frequent condom use, with 28% noting that they used a condom every time and 30.6% reporting condom use almost every time they had sex. Only 9.6% indicated that they never used a condom. The results of hierarchical multiple regression analysis revealed that the perception of quality of general communication with parents, the perception of a partner's attitude towards communication, communication self-efficacy, and communication outcome expectancies, were associated with safer sex communication. However, the association between safer sex communication and condom use was weak, suggesting that other factors excluded from this study are important in determining condom use for this sample of respondents. The findings provide some implications for HIV interventions. Interventions that enhance self-efficacy and positive outcome expectancies related to communication about safer sex are likely to foster discussion with a sexual partner. However, they might not lead to actual condom use. PMID- 11012810 TI - Stress, pre-term labour and birth outcomes. AB - Stress, pre-term labour and birth outcomes Preliminary studies have suggested that stress may be associated with the onset, treatment and outcomes of pre-term labour; however, a systematic comparison of the stress of women with and without pre-term labour has not been reported. Therefore, the purpose of this exploratory study was to compare the stress (daily hassles and mood states) and birth outcomes of black and white women who experienced pre-term labour (PTL) during pregnancy with those who did not. The convenience sample consisted of 35 pregnant women hospitalized in 1996-1997 for the treatment of PTL (24-35 weeks gestation) and 35 controls matched on age, race, parity, gestational age and method of hospital payment. Women in the PTL group had significantly higher tension-anxiety and depression-dejection on the Profile of Mood States (POMS), lower mean birthweight and mean gestational age, and a higher percentage of babies born <37 weeks and weighing 2500 g or less. Black women in the PTL group and white women in the control group had significantly higher scores on the fatigue sub-scale of the POMS and the work and future security sub-scales of the Daily Hassles Scale. Women in the PTL group whose babies weighed 2500 g or less had significantly higher scores on the health, inner concern and financial responsibility sub scales of the Daily Hassles Scale. The findings from this study indicate the need for further exploration of the interaction of race and stress in understanding and preventing PTL and low birthweight and the need to examine the role of social support in preventing pre-term birth after an episode of PTL. PMID- 11012811 TI - Primary health care in Saudi Arabia: applying global aspects of health for all, locally. AB - Primary health care in Saudi Arabia: applying global aspects of health for all, locally This paper describes the application of primary health care principles in the Islamic Kingdom of Saudi Arabia. It arose from a doctoral supervisory experience on a joint programme for women students, operating between a British and Saudi Arabian University. The research looked at nutritional advice given by diploma-level nurses to pregnant women attending primary health care centres in Saudi Arabia. The supervisor supported research that drew on internationally recognized trends in nursing research (the reflexive learner) whilst attending to local requirements and conventions of the culture. The student was encouraged explicitly to site the research within the framework of Islamic teaching and Saudi culture. The Qur'an was used as an overarching framework within which the tenets of primary health care were explored. This was seen to be crucial in addressing World Health Organisation and the International Council of Nurses' views on contextualizing nursing for the greatest benefit of the population. This was of particular relevance in Saudi Arabia where research carried out in the community by women is novel, and as yet there are no nurse theorists from within Saudi culture. PMID- 11012813 TI - Intolerable human suffering and the role of the ancestor: literary criticism as a means of analysis. AB - Intolerable human suffering and the role of the ancestor: literary criticism as a means of analysis This essay explores the experience of intolerable human suffering in Toni Cade Bambara's novel, The Salt Eaters. The method of analysis is literary criticism, a technique that shares many of the same goals as other types of inquiry. It employs close reading to illuminate the novel's meaning(s), thereby revealing information about the nature of intolerable human suffering. Morrison's characteristics of black art is the literary and cultural framework that guides the analysis of Bambara's novel. The paradigm has broad application for nursing. The purpose of this analysis was to describe the role of the ancestral system as a predictor of the trajectory of suffering. The results extend Morrison's paradigm and her notion of ancestor to include traditions and other non-corporeal factors that are essential for well-being and survival. The protagonist in Bambara's novel, Velma Henry, is the patient and exemplar who does not succumb to intolerable suffering because of its cumulative weight, but because she has lost touch with the traditions of her people, an essential component of her ancestral system. The ancestral system is a rich and complex network of individuals, groups, customs and beliefs that are instructive, protective and benevolent. Ancestors are also timeless and provide wisdom, but when the ancestral system is weak or absent, the trajectory of suffering is not favourable. Nurses must learn to recognize intolerable human suffering, to identify the patient's ancestral system, and to work within that system to keep suffering patients from harm. PMID- 11012812 TI - An action research inquiry into a health visitor parenting programme for parents of pre-school children with behaviour problems. AB - An action research inquiry into a health visitor parenting programme for parents of pre-school children with behaviour problems Parent education programmes have recently been given a high profile in this country, such provision deemed necessary because of the commonality of children's behaviour problems and because of their potentially far-reaching effects on the child, the family and society. This action research study, undertaken in Scotland, aims to explore a health visitor parenting programme for parents of pre-school children with behaviour problems. Within this study, children with behaviour problems were identified as those whose parents perceived them to have such problems, the children eliciting a wide range of behaviours and to varying degrees. Critical social science is the chosen methodology, since it allows exploration of how people comprehend their social reality. Group interactions were recorded on audio-tape and themes associated with the methodology of enlightenment, empowerment and emancipation were identified. The results of the study demonstrate how a liberating, collaborative approach to parental education can empower mothers, by increasing understanding and raising self-esteem. PMID- 11012814 TI - On the use of narratives in nursing research. AB - On the use of narratives in nursing research Narratives have always been a path to knowledge in nursing care but are a recent element within nursing research. Therefore, this article deals with the narrative and its use within nursing research. First, the use of narratives in nursing care and nursing research is examined. Second, Paul Ricoeur's narrative theory with its dimensions of interpretation, time, action and ethics is presented as a possible methodological basis. Third, the use of Ricoeur's narrative theory in nursing research is examined, showing that, at present, support mainly comes from Ricoeur's text interpretation theory. Finally, a nursing research approach to the narrative, based on the life-world, is suggested. PMID- 11012815 TI - Using web-based discussion as a teaching strategy: bioethics as an exemplar. AB - Using web-based discussion as a teaching strategy: bioethics as an exemplar Nurses are required to be competent in the area of bioethics based on several accreditation standards. Bioethics courses are one strategy to develop such a competency. Bioethics education itself has various goals including the need to engage in discussion and the ability to debate diverse issues. Providing an educational experience that meets both disciplinary and topic needs can be challenging. On-line conferencing through WebBoard 3.0, a web-based discussion software program, within a graduate level bioethics course successfully contributed to meeting these needs. A technical description of the WebBoard is provided in the following discussion along with a description of the integration of this device into a bioethics course. The WebBoard conferencing allowed all students to enter the class discussion and especially facilitated discussion for reticent speakers and distance learners. Evaluations of the bioethics course from both the faculty and students' perspectives were generally positive. Most students found the technique easy to use and liked the asynchronous format that allowed them to read and contribute at any hour of the day or night. Despite the labour intensive nature of this experience, the strategy ensured participation by every student, a major goal of the project. Developing an interdisciplinary situation would increase the value of such an experience. PMID- 11012816 TI - Moving on from interpretivism: an argument for constructivist evaluation. AB - Moving on from interpretivism: an argument for constructivist evaluation This paper examines the research process in the context of an evaluation of work-based learning. The findings of the evaluation are used to illustrate issues around roles and relationships within interpretivist research where a separation is maintained between the researcher, as investigator, and the participants, as the subject of their investigation. The discussion focuses on: the threatening nature of evaluation and the way in which that affects the process of inquiry and learning; the ways in which people's perceptions of research can act as barriers to the implementation of change; and the consequences of this role separation for practice development. In exploring these issues the paper argues for an approach to evaluation research which (a) emphasizes collaboration, (b) is orientated to change and (c) treats the evaluation process as a learning opportunity through which professionals acquire the skills and knowledge to investigate and advance their own practice. PMID- 11012817 TI - Korean nurses' adjustment to hospitals in the United States of America. AB - Korean nurses' adjustment to hospitals in the United States of America Due to shortage of nurses, more nurses from other countries are employed in health care settings in the United States of America (USA). Little attention has been paid to understanding how culturally different international nurses adjust to USA hospitals. The purpose of this study was to investigate how Korean nurses adjust to USA hospital settings. Grounded theory method was used for sampling procedure, data collection and analysis in order to describe Korean nurses' experiences from their perspective and to develop a substantive theory that explains their process of adjustment. Data were collected using semi-structured formal interviews with a purposive sample of 12 Korean nurses. The interviews were audio-taped and transcribed. Analysis of data, using the constant comparative method, revealed 'adjustment to USA hospitals' as the basic social psychological process. Five categories composed the process: (1) relieving psychological stress; (2) overcoming the language barrier; (3) accepting USA nursing practice; (4) adopting the styles of USA problem-solving strategies; and (5) adopting the styles of USA interpersonal relationships. These five categories capture the essential aspects of the adjustment process and each category contains a set of sub-categories that describe Korean nurses' day-to-day experiences that are critical and also problematic to their adjustment. The process evolves in two stages. In the initial stage, the first three of the five categories greatly influenced the nurses' adjustment. From the perspective of the nurses in the study, the initial stage lasts about 2 to 3 years. The remaining two categories are principal components of the later stage. It takes an additional 5 to 10 years to complete this stage. This model highlights both distress and accomplishments of Korean nurses during their adjustment to USA hospitals. The results of the study may help USA nurses gain insight in designing and implementing orientation programmes to facilitate and support Korean nurses' adjustment to USA hospitals. PMID- 11012818 TI - An impossible dream? images of nursing held by pre-registration students and their effect on sustaining motivation to become nurses. AB - An impossible dream? Images of nursing held by pre-registration students and their effect on sustaining motivation to become nurses Each year approximately 13 000 people enter higher education programmes leading to nurse registration in England. Evidence suggests that students enter nursing with strong images about how they will practice. This paper explores the nature of such images and how they are used to inform students' approaches to nursing practice. Findings come from a longitudinal study designed to investigate how students develop their professional knowledge whilst working in clinical settings. Eight pre registration degree-course nursing students participated in the study. A multi method approach to data collection was used over their 4-year programme. One involved in-depth interviews taking place during each of students' clinical placements. Data were analysed manually and subjected to a constant comparative method of analysis. From this material individual case studies of the five completing students were constructed, with participants checking their own case study to ensure that it reflected their intended meaning. A second phase followed where cross-case comparison addressed each of the original research questions. The question relevant to this paper was: What were students' conceptions of nursing on entry, and how do these influence their development? Findings indicate that participating students' preconceptions of nursing had a profound influence on their decision whether to continue with their course, sometimes despite social and academic set-backs, or to leave nursing. An important contribution to realizing their aims was their supernumerary status and effective support from knowledgeable and experienced practitioners. These findings indicate that with better understanding of the relationship between this form of personal knowing and practice, educators would have more information with which to select students and to design professional curricula. PMID- 11012819 TI - Social perceptions of cancer and their impacts: implications for nursing practice arising from the literature. AB - Social perceptions of cancer and their impacts: implications for nursing practice arising from the literature At the millennium cancer still holds a special mystique and is imbued with socio-cultural meanings, which extend far beyond the rational, scientific and biological facts of the disease. Excessive fear and dread may cause family and friends to display avoidance or overprotective behaviours to the ill person, who may subsequently perceive dissatisfaction with social support. Drawing on a literature review this paper explores the impact of cancer on social relationships. Interpersonal strain in relationship is often explained in the stigmatization of the illness and this concept is explored through contemporary social theorizing. These findings have direct implications for nursing practice where the goal of care is to enhance the support relationship. PMID- 11012820 TI - Towards an aesthetics of nursing. AB - Towards an aesthetics of nursing This paper re-appraises the work of Barbara Carper on aesthetics and the art of nursing. It identifies serious flaws in Carper's original arguments and the way in which she and subsequent authors have conflated the concepts of art and aesthetics in nursing. The paper explores a broader approach to aesthetics and proposes a way in which a theoretical approach to nursing aesthetics could be developed. The paper concludes that nursing is a fit object of aesthetic appreciation and that aesthetic quality is a necessary attribute of good nursing practice. PMID- 11012821 TI - Do parents or surrogates have the right to demand treatment deemed futile? An analysis of the case of Baby L. AB - Do parents or surrogates have the right to demand treatment deemed futile? An analysis of the case of Baby L The purpose of this discussion paper is to address and analyse the ethical issues arising from the following questions: Do parents or surrogates of newborn infants have the right to demand treatment deemed 'futile'? Should the religious beliefs of the infant's parents be given special consideration when deciding on the correct course of action? The case of Baby L, an infant born with severe disability will be used to aid the analysis. It is argued that health care providers have no obligation to provide 'futile' treatment based on the surrogates' right to autonomy, but an obligation may arise from a duty of benevolence. However, acting from a duty of benevolence can ignore considerations of justice and fairness and does not always prompt the right course of action. Any decision regarding treatment options will involve ranking the beliefs and values of the parents or surrogate against the integrity of the health care team and the interest of society as a whole. The consequences of continuing or discontinuing life-sustaining treatment for the infant, the parents and the health care team will also be considered and examined. PMID- 11012822 TI - Forthcoming contents of volume 32, number 4, october 2000 PMID- 11012823 TI - Digital imaging in transmission electron microscopy. AB - The digital revolution currently under way, as evidenced by the rapid development of the Internet and the world-wide-web technologies, is undoubtedly impacting the field of transmission electron microscopy (TEM). Digital imaging systems based on charge-coupled device (CCD) technologies, with pixel array size up to 2 k x 2 k at the present and increasing, are available for TEM applications and offer many attractions. Is it time to phase out film cameras on TEMs and close the darkrooms for good? This paper reviews digital imaging technologies for TEM at different voltages, and contrasts the performance of digital imaging systems with that of TEM film. The performance characteristics of CCD-based digital imaging systems, as well as methods for assessing them, are discussed. Other approaches to digital imaging are also briefly reviewed. PMID- 11012824 TI - Two-photon image correlation spectroscopy and image cross-correlation spectroscopy. AB - We introduce two-photon image correlation spectroscopy (ICS) using a video rate capable multiphoton microscope. We demonstrate how video rate two-photon microscopic imaging and image correlation analysis may be combined to measure molecular transport properties over ranges typical of biomolecules in membrane environments. Using two-photon ICS, we measured diffusion coefficients as large as 10(-8) cm2 s(-1) that matched theoretical predictions for samples of fluorescent microspheres suspended in aqueous sucrose solutions. We also show the sensitivity of the method for measuring microscopic flow using analogous test samples. We demonstrate explicitly the advantages of the image correlation approach for measurement of correlation functions with high signal-to-noise in relatively short time periods and discuss situations when these methods represent improvements over non-imaging fluorescence correlation spectroscopy. We present the first demonstration of two-photon image cross-correlation spectroscopy where we simultaneously excite (via two-photon absorption) non-identical fluorophores with a single pulsed laser. We also demonstrate cellular application of two photon ICS for measurements of slow diffusion of green fluorescent protein/adhesion receptor constructs within the basal membrane of live CHO fibroblast cells. PMID- 11012825 TI - Stereological estimation of integral mixed curvature with application AB - This article addresses a bounded system of compact smooth surfaces in three dimensional space. Recently, an unbiased estimator of the integral mixed curvature of the given system based on a vertical sampling design has been proposed. The aims of the present paper are: (i) to show that the proposed estimator may have an infinite variance; (ii) to suggest a modification that has better statistical properties; (iii) to extend the point estimator to a function that monitors the curvature along the gradient microstructures; (iv) to present an application of the method to real microscopic images. PMID- 11012826 TI - Testing local dependence of spatial structures on images AB - Associations between two spatial processes can be due to a real dependence between the two processes or to the dependence on common underlying variables. We propose to test the existence of a real dependence by use of local tests, leading to a global test of real dependence and a map of local interactions. We present first how classical interaction tests based on random rotations between completely observed processes such as those developed by Berman (Berman. Appl. Statist. (1986) 35, 54-62), can be integrated in local analyses. For this purpose, tests are first performed locally, and the distribution of their p values is then compared to the corresponding value under the null hypothesis. A similar approach is proposed to test non-stationarity of a point pattern by using distance statistics popularized by Diggle (Diggle. Statistical Analysis of Spatial Point Patterns. (1983) Academic Press, New York). The problem of testing the interaction between a random field and a censoring area pattern process is discussed and an approach similar to the preceding ones is then proposed. The methods are mainly applied to agricultural examples but they can be applied to any microscopical images for which one wishes to analyse the spatial structure. PMID- 11012827 TI - Is it reasonable to assume a uniformly distributed cooling-rate along the microslide of a directional solidification stage? AB - It is commonly assumed that the cooling-rate along the microslide of a directional solidification stage is uniformly distributed, an assumption which is typically applied in low cooling-rates studies. A new directional solidification stage has recently been presented, which is specified to achieve high cooling rates of up to 1.8 x 104 degrees C min-1, where cooling-rates are still assumed to be uniformly distributed. The current study presents a closed-form solution to the temperature distribution and to the cooling-rate in the microslide. Thermal analysis shows that the cooling-rate is by no means uniformly distributed and can vary by several hundred percent along the microslide in some cases. Therefore, the mathematical solution presented in this study is essential for experimental planning of high cooling-rate experiments. PMID- 11012828 TI - Cathodoluminescence microscopy characterization of chrome-free refractories for copper smelting and converting furnaces AB - The results of an experimental program assessing the potential of several chrome free refractory materials as potential replacements for the mag-chrome brick currently used in copper production furnaces are presented. Several commercial chrome-free bricks were subjected to the standard dip test in a high-copper calcium ferrite slag. The mineralogical changes in the bricks resulting from their interaction with the molten slag are described. The use of optical cathodoluminescence microscopy as an analytical tool is highlighted, along with reflected light microscopy and scanning electron microscopy-energy dispersive spectroscopy. Using the three tools together provides a description of the experimental results not achievable using one or two alone. The penetration resistance of the potential replacement refractories is comparable to that of mag chrome, but the substitution of Fe2+ for Mg2+ in the periclase and spinel crystalline structures of the replacements reduces their corrosion resistance compared with mag-chrome, diminishing the likelihood that they will serve as reliable alternatives. PMID- 11012829 TI - Electric field depolarization in high aperture focusing with emphasis on annular apertures AB - Electromagnetic focusing theory of light predicts that at high apertures field components arise that are polarized perpendicular to the initial polarization. Although vectorial depolarization has received considerable attention in focusing theory, no evidence has been presented as to its relevance in experiments. We measure the intensity of the perpendicularly orientated field in the focal region by utilizing monomolecular, fluorescent polydiacetylene layers whose transition dipoles are orientated in a single direction. For a 1.4 numerical aperture oil objective lens illuminated with linearly x-polarized light, we find that the integral of the modulus squared of the y-polarized focal field amounts to 1.5% of its x-polarized counterpart. In particular, we show here that the depolarization increases when using annular apertures. Annuli formed by a central obstruction with a diameter of 89% of that of the entrance pupil raise the integral to 5.5%. This compares well with the value of 5.8% predicted by electromagnetic focusing theory; however, the depolarization is also due to imperfections connected with focusing by refraction. Besides fluorescence microscopy and single molecule spectroscopy, the measured intensity of the depolarized component in the focal plane is relevant to all forms of light spectroscopy combining strong focusing with polarization analysis. PMID- 11012830 TI - Strategies for the compensation of specimen-induced spherical aberration in confocal microscopy of skin. AB - We consider various strategies for confocal imaging of human skin which seek to reduce the effects of the specimen-induced aberrations. We calculate the spherical aberration introduced by the stratified structure of skin and show how the confocal signal is affected when attempting to image at various depths within the dermis. Using simple methods it is shown how images might be improved by compensating for the induced aberration. The methods include the use of an iris to reduce the pupil area, changing the refractive index of the immersion medium and using a lens with variable coverglass correction. PMID- 11012831 TI - A two-channel four-dimensional image recording and viewing system with automatic drift correction. AB - Four-dimensional image acquisition systems have been described to analyse various developmental processes, for example, the Caenorhabditis elegans cell lineage. A practical problem that is often encountered during recordings is mechanical slippage of the microscope stage, causing the sample to drift out of focus. Furthermore, with the advent of green fluorescent protein (GFP) as an in vivo marker, affordable two-channel imaging systems are needed to correlate gene expression with changes through development. To overcome the mechanical drift a device-independent, software-only solution for the MacOS was devised that can compensate for Z-axis drifts in sample position. The software also allows recording of 4D stacks in two channels. To correct for drift, a small reference object beside the main object to be recorded is kept in focus using a simple autofocus principle, and this automatic drift correction allows for effective 4D recordings. In addition to the Z-axis drives and the shutters of the microscope, a video camera can be computer controlled to switch between two light levels. Second channel live GFP recordings are presently limited by the fact that the high intensity of the blue light heats and kills C. elegans embyros quickly. To view and annotate the stacks a MacOS viewing application was developed. PMID- 11012833 TI - Neuroendocrinology briefings 10: biological timekeeping PMID- 11012832 TI - Neuroendocrinology briefings 10: biological timekeeping. AB - The existence of biological clocks was once the subject of considerable debate. However, research has confirmed that a 24 h (circadian) clock in our brain has a pronounced influence on when we sleep, eat, drink, etc. Disruptions in clock function compromise normal daily patterns of hormone release, sleep and body temperature, and can affect mental and physical well-being. The recent identification of clock genes and the neural pathways to the clock will facilitate the development of effective strategies to treat disorders of circadian function. PMID- 11012834 TI - Ultradian corticosterone rhythm and the propensity to behave aggressively in male rats. AB - Ultradian fluctuations in plasma glucocorticoids have been demonstrated in a variety of species including humans. The significance of such rhythms is poorly known, although disorganized ultradian glucocorticoid rhythms have been associated with behavioural disorders. Here we report that ultradian glucocorticoid rhythms may establish the propensity to behave aggressively in male rats. Male rats were significantly more aggressive in the increasing phase of their corticosterone fluctuation when confronting a male intruder than counterparts in the decreasing phase of their corticosterone fluctuations facing such opponents. Corticosterone fluctuations were mimicked by a combination of treatments with the corticosterone synthesis inhibitor metyrapone and corticosterone. Again, males with increased plasma corticosterone levels were more aggressive than counterparts with a decreased plasma corticosterone concentration. These data suggest that the behavioural response to an aggressive challenge may vary in the same animal across the day due to the pulsating nature of corticosterone secretion. Aggressive behaviour is also episodic in humans; moreover, intermittent explosive behaviour is recognized as a psychological disorder. It can be hypothesized that a temporal coincidence between the occurrence of a challenge and a surge in plasma corticosterone concentration may be one of the factors that promote episodic aggressive outbursts. PMID- 11012835 TI - Basic fibroblast growth factor priming increases the responsiveness of immortalized hypothalamic luteinizing hormone releasing hormone neurones to neurotrophic factors. AB - The participation of growth factors (GFs) in the regulation of luteinizing hormone releasing hormone (LHRH) neuronal function has recently been proposed, but little is known about the role played by GFs during early LHRH neurone differentiation. In the present study, we have used combined biochemical and morphological approaches to study the ability of a number of GFs normally expressed during brain development, including basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I) to induce survival, differentiation, proliferation, and phenotypic expression of immortalized (GT1-1) LHRH neurones in vitro, at early (3-days in vitro, 3-DIV) and late (8-DIV) stages of neuronal differentiation. Comparison of GF-treated vs untreated neurones grown in serum-deprived (SD) medium demonstrated bFGF to be the most potent, and insulin the least active in promoting neuronal differentiation. Thus, at both 3-DIV and 8-DIV, but especially at 8-DIV, bFGF induced the greatest increase in the total length and number of LHRH processes/cell and in growth cone surface area. bFGF was also the most active at 3-DIV, and IGF-I at 8-DIV, in counteracting SD-induced cell death, whereas EGF was the most potent in increasing [3H]thymidine incorporation. All GFs studied decreased the spontaneous release of LHRH from GT1-1 cells when applied at 3-DIV or 8-DIV, except for insulin which was inactive at both time-points and bFGF which was inactive at 8-DIV. Pre-treatment of GT1-1 cells with a suboptimal ('priming') dose of bFGF for 12 h followed by application of the different GFs induced a sharp potentiation of the neurotrophic and proliferative effects of the latter and particularly of those of IGF-I. Moreover, bFGF priming counteracted EGF-induced decrease in LHRH release and significantly stimulated LHRH secretion following IGF-I or insulin application, suggesting that bFGF may sensitize LHRH neurones to differentiating effects of specific GFs during development. PMID- 11012836 TI - Hypervascularization in the magnocellular nuclei of the rat hypothalamus: relationship with the distribution of aquaporin-4 and markers of energy metabolism. AB - In the magnocellular nuclei of the hypothalamus, there is a rich vascular network for which the function remains to be established. In the supraoptic nucleus, the high vascular density may be one element, which together with the water channel aquaporin-4 expressed in the astrocytes, is related to a role in osmoreception. We tested the osmoreception hypothesis by studying the correlation between vascular and cellular densities in the paraventricular nucleus and the supraoptic nucleus. Whether aquaporin-4 is likely to contribute to osmoreception was tested by studying the distribution in the magnocellular nuclei of the hypothalamus. The high vascular density may also reflect a high metabolic activity due to the synthesis of vasopressin and oxytocin. This metabolic hypothesis was tested by studying the regional cytochrome oxidase histochemistry, the local cerebral blood flow, and the density of glucose transporter type-1 in the supraoptic and paraventricular nuclei. All the magnocellular nuclei were characterized by an extended and intense aquaporin-4 labelling and a weak cytochrome oxidase histochemistry. The highest vascular density was found in the supraoptic nucleus and the magnocellular regions of the paraventricular nucleus. The local cerebral blood flow rates were surprisingly low in the paraventricular nucleus and the supraoptic nucleus in comparison to the cerebral cortex. Furthermore in these nuclei, the antibody for glucose transporter type-1 revealed two populations of vessels differing by their labelling intensity. The similarities observed between the different nuclei suggest that, in the hypothalamus, all magnocellular regions sense the plasma osmolarity. The low local cerebral blood flow, and the patterns of glucose transporter type-1 labelling and cytochrome oxidase histochemistry suggest that the high vascularization of these hypothalamic nuclei is not related to a high metabolic capacity in basal conditions. PMID- 11012837 TI - The effects of ionotropic agonists of excitatory amino acids on the release of arginine vasopressin in rat hypothalamic slices. AB - The effects of ionotropic excitatory amino acids agonists on the release of vasopressin from rat hypothalamic slices were studied. Incubation with increasing doses of NMDA, kainate or AMPA decreased the release of vasopressin in a dose dependent manner. The values of the IC50 were 1.0, 9.6, or 3.7 x 10-8 M, respectively. The inhibitory effect of the various excitatory amino acids tested was blocked by coincubation with their respective antagonists. Vasopressin secretion was stimulated to 140.3 +/- 7.6% of controls when the slices were obtained from chronically (7 days) salt-loaded rats. Addition of 1 x 10-7 M NMDA or 1 x 10-6 M kainate to the incubation medium antagonized the salt loading induced increase in vasopressin release. Incubation with 1 x 10-4 M tetrodotoxin did not change basal vasopressin release, but it blocked the decrease in vasopressin secretion induced by 1 x 10-7 M NMDA or 1 x 10-6 M kainate or 1 x 10 6 M AMPA. Incubation with 1 x 10-5 M phaclophen (a GABAB antagonist) and 1 x 10-5 M bicuculline (a GABAA antagonist) was without effect on basal vasopressin secretion while it reversed the inhibition of vasopressin release induced by 1 x 10-7 M NMDA. Incubation with 1 x 10-6 M GABA alone decreased vasopressin secretion to 64.6 +/- 6.9% of control values. The inhibitory effect of GABA did not change when 1 x 10-7 M NMDA was added to the incubation medium. These findings demonstrate that ionotropic excitatory amino acids agonists inhibit vasopressin secretion from hypothalamic slices. They strongly suggest that this inhibitory effect is mediated through local GABAergic interneurones. PMID- 11012838 TI - Agouti related protein in the rat adrenal cortex: implications for novel autocrine mechanisms modulating the actions of pro-opiomelanocortin peptides. AB - Agouti related protein (AgRP) is a recently discovered melanocortin receptors (MCR) antagonist implicated in the control of feeding behaviour. Expression of AgRP has been shown to be localized by in situ hybridization to the arcuate nucleus and median eminence of the brain, where it acts as an antagonist to the MC3 and MC4 receptors, while in the periphery the only significant expression was located in the adrenal medulla. As AgRP is only a weak antagonist of the MC2 and MC5 receptors, which are expressed principally by adipocytes and in the adrenal cortex, the question arizes as to the function of peripheral AgRP. In this study, we investigated the expression of AgRP in the rat adrenal and suggest that it is expressed in the adrenal cortex and not as previously described in the medulla. We also show that AgRP mRNA expression is upregulated in the adrenal during fasting and in the contralateral gland following unilateral adrenalectomy but not during chronic stress. These results indicate an as yet undefined role for AgRP in the periphery and are supportive of the suggestion that a further melanocortin receptor exists. PMID- 11012839 TI - Pharmacological evidence that prolactin acts from late gestation to promote maternal behaviour in rabbits. AB - We investigated the role of prolactin and suckling stimulation in the expression of maternal behaviour of primiparous rabbits. Bromocriptine (1 mg/kg/day), given to intact mothers across postpartum days 1-5, decreased serum concentrations of prolactin to undetectable levels, reduced crouching, and increased time inside the nest. Failure of maternal nest-building, provoked by bromocriptine injections from pregnancy day 26 to parturition or to postpartum day 5, correlated with a stronger reduction in crouching and an increased time inside the nest, measures of disturbed maternal behaviour, on postpartum days 3 and 5. Preventing suckling by thelectomy did not prevent prolactin release but reduced crouching incidence and increased the time spent inside the nest on postpartum days 3 and 5. Bromocriptine, injected in thelectomized mothers across postpartum days 1-5, further reduced the incidence of crouching and increased the time spent inside the nest on postpartum days 3 and 5. We conclude that prolactin acting prepartum facilitates maternal behaviour initiation in rabbits and, together with pup stimulation, maintains this behaviour across lactation. PMID- 11012840 TI - Calcium-independent, tyrosine phosphorylation-dependent effects of serum on the morphology of cultured neurohypophysial astrocytes. AB - Activation of adenylate cyclase induces cultured neurohypophysial astrocytes (pituicytes) to change from a protoplasmic, nonstellate form to a stellate form. Stellation is inhibited and reversed (destellation) by serum. The objective of the present studies was to examine the roles of Ca2+ and tyrosine phosphorylation in mediating these morphological changes. The effects of forskolin (to induce stellation) and serum (to inhibit and reverse stellation) were not affected by replacement of Ca2+ with Co2+ in the medium or by treatment of cultures with thapsigargin. However, genistein, a specific inhibitor of tyrosine kinase(s), significantly reduced the effect of serum on forskolin-induced stellation. Also, dephostatin, a specific inhibitor of tyrosine phosphatase, inhibited forskolin induced stellation. In contrast, genistein did not have a dramatic effect on serum-induced destellation. The data demonstrate that morphological changes exhibited by cultured pituicytes are independent of Ca2+ but may be modulated by the activity of tyrosine kinase(s) and phosphatase(s). PMID- 11012841 TI - Effect of suckling on NADPH-diaphorase (Nitric oxide synthase, NOS) reactivity and NOS gene expression in the paraventricular and supraoptic nuclei of lactating rats. AB - This study examined the effect of suckling on nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d, a histochemical marker for nitric oxide synthase, NOS) reactivity and neuronal NOS mRNA expression in the paraventricular (PVN) and supraoptic (SON) nuclei of lactating rats. Freely nursing (non-separated) dams and those separated from pups for 12 h and then reunited for 0, 15, 30, 60, 90, 120 and 180 min were used for the study. Dams separated from pups and sacrificed at time zero (without reunion) showed a significant decrease in NADPH-d staining and NADPH-d positive cells as well as in the NOS mRNA expression in the PVN and SON compared to that observed in non-separated dams. Reunion with pups and restoration of suckling significantly increased NADPH-d reactivity after 15, 30, 60 min, but not after 90, 120 and 180 min compared to non-reunited pups-deprived dams. A pattern of NADPH-d reactivity and neuronal NOS mRNA expression indistinguishable from that observed during free lactation was reinstated shortly (15 min) after the restoration of suckling stimulus, suggesting that the NADPH-d reactivity in lactation depends on the presence of the suckling stimulus. These results show that suckling stimulus may play a modulatory role in the regulation of NOS reactivity in the magnocellular neurones of the hypothalamic PVN and SON during lactation. PMID- 11012842 TI - The effects of ciliary neurotrophic factor on the hypothalamo-pituitary gonadal axis in vitro in female rats. AB - Ciliary neurotrophic factor (CNTF) is a member of the neuropoietic family of cytokines. CNTF exerts its actions through activation of a receptor complex, which shows similarity of sequence, second messenger systems and distribution to the leptin receptor. Leptin has been demonstrated to exert profound effects on the hypothalamo-pituitary gonadal axis. This study examines the in vitro effects of CNTF on hypothalamic luteinizing hormone releasing hormone release (LHRH) and pituitary luteinizing hormone (LH) release compared to those of leptin in the female. We report that CNTF stimulates LHRH release from medial basal hypothalamic explants harvested from proestrous female rats and this effect is of similar magnitude to that seen with leptin. In contrast, CNTF suppresses LHRH stimulated LH release from dispersed anterior pituitary cells harvested from proestrous female rats but has no effect on basal LH release. Leptin stimulates basal LH release but has no effect on LHRH-stimulated LH release. The suppressive effect of CNTF on LHRH-stimulated LH release has been confirmed in perifused anterior hemipituitaries. These results suggest a differential effect of CNTF on the hypothalamo-pituitary gonadal axis and a possible role in the modulation of pituitary gonadal function. PMID- 11012844 TI - Selective corticotropin-releasing factor type 1 receptor antagonist blocks conditioned fear-induced release of noradrenaline in the hypothalamic paraventricular nucleus of rats. AB - The effects of conditioned fear on the release of noradrenaline in the hypothalamic paraventricular nucleus (PVN) and the involvement of corticotropin releasing factor (CRF) receptor type 1 (CRFR1) in conditioned fear-induced changes in noradrenaline release were examined by intracerebral microdialysis in rats. Conditioned fear was produced by placing animals into a box where they had previously been exposed to a 5-min period of electric footshock, 135 min prior to the start of experiment. Conditioned fear for 20 min produced a significant increase in the release of noradrenaline in the PVN. Intraperitoneal preadministration of a selective nonpeptidic CRFR1 antagonist, CRA1000, completely blocked the conditioned fear-induced release of noradrenaline. These results suggest that CRFR1 is involved in the release of noradrenaline in the hypothalamic PVN induced by conditioned fear. PMID- 11012843 TI - Blunted pituitary-adrenocortical stress response in adult rats following neonatal dexamethasone treatment. AB - Glucocorticoids have a prominent impact on the maturation of the stress-related neuroendocrine system and on the postnatal establishment of adaptive behaviour. The present study aimed at investigating the stress responsiveness of the hypothalamo-pituitary-adrenocortical (HPA) axis in young and adult rats after neonatal treatment with the synthetic glucocorticoid agonist, dexamethasone. Newborn male Wistar rats were injected s.c. with 1 microg/g dexamethasone on postnatal days 1, 3 and 5. Circulating adrenocorticotropic hormone (ACTH) and corticosterone concentrations were measured in the resting state and following a 30-min cold stress at the age of 10 days, as well as after a 30-min restraint stress at the age of 14 weeks. Also in adults, pituitary and adrenocortical hormone responsiveness was evaluated after i.v. administration of 2 microg/kg corticotropin releasing hormone (CRH). In addition, glucocorticoid (GR) and mineralocorticoid receptor (MR) binding capacities were assessed in the pituitaries of adult rats. The results showed that at day 10 basal ACTH concentration was elevated while the cold stress-evoked ACTH response was attenuated in the dexamethasone-treated rats. As adults, treated rats showed a suppressed elevation of both ACTH and corticosterone plasma concentrations in response to restraint, while basal hormonal concentrations were not altered. There was no difference in the magnitude of the CRH-induced elevation of ACTH and corticosterone concentrations initially; however, the dexamethasone-treated animals showed a prolonged secretion of both hormones. These animals also showed a selective decrease in pituitary GR binding capacity. Neonatal dexamethasone treatment strongly suppressed body weight gain, and adrenal and thymus weights in the early phase of postnatal development. By adulthood, the body and adrenal weights were normalized while thymus weight was greater than in controls. These findings indicate that neonatal dexamethasone treatment permanently alters HPA axis activity by reducing stress responses to cold and restraint probably through supra-pituitary actions, and by decreasing the effectiveness of feedback through a diminished GR binding in the pituitary. PMID- 11012845 TI - Glutamatergic regulation of gonadotropin releasing hormone mRNA levels during development in the mouse. AB - The aims of these studies were to investigate the time course of the increase in gonadotropin releasing hormone (GnRH) mRNA levels during sexual development in the mouse, and to test the hypothesis that the neurotransmitter glutamate regulates the GnRH secretory system via actions at the level of GnRH gene expression. GnRH mRNA abundance was estimated by measuring silver grains generated by in situ hybridization of an 35S-labelled oligonucleotide probe. There was a significant increase in GnRH mRNA abundance between the day of birth (P0) and postnatal day 2 (P2) in male mice, but no further increases at later ages when overt pubertal changes are manifest. GnRH mRNA levels also increased significantly between P0 and P2 in female mice. Treatment with the glutamate agonist NMDA caused a significant increase in GnRH mRNA levels in neonatal (P0) mice and adult male mice within 30 min of treatment, which is consistent with previous studies in the rat implicating glutamate in the regulation of GnRH mRNA stability. Treatment with the glutamate antagonist CGP40116 caused an equally rapid decrease in GnRH mRNA levels in adult mice and in mice on P5 after the neonatal increase in GnRH gene expression, but was without effect in mice on P0, prior to the developmental increase. These observations that the effect on GnRH mRNA levels of blocking endogenous glutamatergic signalling depends upon the developmental stage suggest that endogenous glutamate maintains GnRH mRNA levels in adult mouse, and is a potential regulator of the developmental increase seen in the neonatal period. PMID- 11012846 TI - Selective blockade of the mineralocorticoid receptor impairs hypothalamic pituitary-adrenal axis expression of habituation. AB - The present study investigated the role of mineralocorticoid receptors (MR) and glucocorticoid receptors (GR) in the expression of habituation of the hypothalamic-pituitary-adrenal (HPA) axis response to stress. Male rats were restrained for 1 h per day for six consecutive days. On day 6, 1 h prior to restraint stress, both restraint-naive and repeatedly restrained rats were injected s.c. with either vehicle (propylene glycol) or one of three corticosteroid receptor antagonist treatments: selective MR antagonist (RU28318 or spironolactone), selective GR antagonist (RU40555), or both MR and GR antagonists combined (RU28318 + RU40555). Blood samples were collected for corticosterone measurement at the beginning of stress, during stress, and 1 h after stress termination. Repeated restraint stress produced significant habituation of corticosterone responses. Acute treatment with the combined MR and GR antagonists prevented the expression of habituation. When tested alone, the MR antagonist also blocked the expression of corticosterone-response habituation, whereas the GR antagonist had no effect. Neither the MR, nor the GR antagonists alone, significantly altered the corticosterone response to restraint in rats exposed to restraint for the first time. The final experiment examined the corticosterone response to a corticotropin releasing hormone (CRH, 3 microg/kg i.p.) challenge. Neither previous exposure to restraint or acute pretreatment with the combined MR and GR antagonists (RU28318 + RU40555) altered the corticosterone response to CRH challenge. This result indicates that the expression of habituation and its blockade by corticosteroid receptor antagonists is not a result of altered pituitary-adrenal response to CRH. Overall, this study suggests that MR plays an important role in constraining the HPA axis response to restraint stress in restraint-habituated rats. The dependence of the HPA axis on MR-mediated corticosteroid negative feedback during acute stress may be an important mechanism that helps maximize the expression of stress habituation and thereby minimize exposure of target tissues to corticosteroids in the context of repeated stress. PMID- 11012847 TI - Drooling: review of the literature and proposals for management. AB - The aetiology of drooling is described alongside the alternative treatment strategies for what is a disturbing disorder for many children. The advantages and disadvantages of each treatment modality are discussed. Conditions for optimal management are described. PMID- 11012848 TI - Three-dimensional force measurements on oral implants: a methodological study. AB - This paper describes a methodology that allows in vitro and in vivo quantification and qualification of forces on oral implants. Strain gauges are adapted to the outer surface of 5.5 and 7 mm standard abutments (Branemark System, Nobel Biocare, Sweden). The readings of the strain gauges are transformed into a numerical representation of the normal force and the bending moment around the X- and Y-axis. The hardware and the software of the 3D measuring device based on the strain gauge technology is explained and its accuracy and reliability tested. The accuracy level for axial forces and bending moments is 9.72 N and 2.5 N x cm, respectively, based on the current techniques for strain gauged abutments. As an example, an in vivo force analysis was performed in a patient with a full fixed prosthesis in the mandible. Since axial loads of 450 N and bending moments of 70 N x cm were recorded, it was concluded that the accuracy of the device falls well within the scope of our needs. Nevertheless, more in vivo research is needed before well defined conclusions can be drawn and strategies developed to improve the biomechanics of oral implants. PMID- 11012849 TI - Stress distribution in the temporomandibular joint affected by anterior disc displacement: a three-dimensional analytic approach with the finite-element method. AB - The purpose of this study was to investigate the influences of anterior disc displacement on TMJ loading during maximum clenching by use of finite-element analysis. Based on a young human dry skull, an analytic model of the mandible including the TMJ was developed. In addition to the standard model with normal disc-condyle relation, two models were designed to simulate various degrees of anterior disc displacement. In the standard model, compressive stresses were induced in the anterior, middle and lateral areas on the condyle and glenoid fossa, whereas tensile stresses were observed in the posterior and medial regions. In the models with anterior disc displacement, compressive stresses were recognized in all the areas of TMJ components excluding the bilaminar zone. Shear stresses in the articular disc and bilaminar zone significantly increased in most areas. In conclusion, stress distributions in the TMJ with a normal disc position was substantially different from those with anterior disc displacement, suggesting that the progress in disc displacement may have some association with the nature of stress distributions in the TMJ, in the articular disc in particular. PMID- 11012850 TI - Sectional prosthesis with hollow obturator portion made of thin silicone layer over resin frame. AB - Magnetically retained sectional obturator prostheses, consisting of a hollow obturator portion made of thin silicone layer over a resin frame and a denture portion, were fabricated to alleviate some of the limitations of silicone materials, and applied to two edentulous patients with maxillectomy defects. The silicone obturator allowed profound engagement of undercuts within the defect, resulting in adequate retention, support and stability of the prostheses for over 3 years. The patients have achieved improvement in speech and mastication without complications, by use of the obturator prostheses. PMID- 11012851 TI - Bolus dimensions in normal chewing. AB - Samples of 17 different types of chewing gum weighing between 0.3 and 22 g were presented to eight subjects. After chewing the samples for between 20 and 100 strokes the boluses of gum were removed from the mouth and their length was measured. Bolus length increased with weight from 0.3 to 4 g, but remained constant between 4 and 18 g at which point it began to increase once more. The range of sizes associated with this plateau is similar to the range of sizes of natural bites with other foods. PMID- 11012852 TI - Evaluation of stresses caused by dentin pin with finite elements stress analysis method. AB - The aim of the present study was to show the dimensions and the amount of stresses caused by pins on dentin. Mathematically modelled stainless steel and titanium pins were applied to mandibular first molar teeth with extensive crown destruction. The stress caused by the pins was examined with the finite elements method (FEM). In both types of pin, the maximum diffuse and the dense stress areas were located at the bottom of the pin channel. It is believed that these stresses should be taken into consideration when evaluating the advantages and disadvantages of pin application to teeth with destroyed crowns. PMID- 11012853 TI - Acoustic emissions generated in aged dental composites using a laser thermoacoustic technique. AB - The heating up of dental composites by laser will produce acoustic emissions (AEs) that may be related to fracture mechanisms in the composites. It has been proved that the mechanical properties of dental composites are affected by storage in food simulating liquids, i.e. 75% ethanol, which has a solubility parameter approximating to that of bisphenol glycidyl dimethacrylate (BisGMA) resin. A new method was innovated to evaluate the laser-induced AEs in dental composites aged by 75% ethanol solution. Model systems (50/50 BisGMA/TEGDMA resin filled with 0% and 75 wt.% 5-10 microm silanized BaSiO6) as well as three commercial composites (Marathon One, Z100 and Herculite XRV) were used in this study. Nine samples acting as the control group were tested to establish the correlation of AEs to laser power. The effect of ageing by immersion in 75% ethanol on AEs and diametral tensile strength (DTS) was then evaluated. A quasi continuous wave CO2 laser was used to heat up the composites. AEs of frequency 100-200 kHz were collected, filtered, recorded and processed using a 4610 Smart Acoustic Monitor. Burst patterns, which formally were assumed to be correlated to fracture mechanisms, were also identified from the data obtained at laser power > or = 5 W for commercial composites and > or = 4 W for model systems. Higher laser powers cause the AE to increase for all composites except unfilled model resin. AEs as a function of power for all aged systems were flat (< 100 events) below 4 W. Emissions then rose sharply to > 1000 events at 7.1 W. Statistically significant differences were found between the AEs obtained at 5 W (commercial composites) and those at 4.3 W (model systems) for material systems and storage times. Marathon One was less affected by the laser and an abrupt change in AE was found between days 0 and 7 of storage for all commercial composites. The AE value from the unfilled model resin was found to be significantly different from that of the model composites. However, they showed an increase in AEs with length of storage time, which was inversely associated with the decreased tendency of their immersed DTS values. Laser-induced AEs may be a valuable adjunct to conventional mechanical testing. PMID- 11012854 TI - In vivo measurement of colour changes in natural teeth. AB - It has been observed that teeth become lighter when they are dried. The present study was designed to quantify these changes and the time taken for tooth colour to return to normal. The colour of an upper central incisor in each of seven subjects was measured using a reflectance spectrophotometer before and after application of a rubber dam and, in another seven subjects before and after taking a polyvinylsiloxane impression. There were statistically significant changes in the L*, a* and b* values following rubber dam application and in the L* value following impression taking. The results demonstrate that teeth become brighter and less colour saturated after rubber dam application and brighter after impression taking. The original values were regained after 30 min. PMID- 11012855 TI - Influence of alteration of occlusal relationship on activity of jaw closing muscles and mandibular movement during submaximal clenching. AB - The aim of this study was to investigate the relationships between occlusal contacts, responses of muscles, and jaw movements during simulated clenching. Seven healthy human males who possessed complete natural dental arches with normal occlusion, ranging from 24 to 29 years of age, volunteered for this study. Acrylic occlusal stops were fabricated for the lower jaw to simulate various occlusal conditions. Vertical movements of the lower jaw were measured by four sets of linear variable differential transformers. Simultaneously, electromyographic (EMG) activity from the bilateral masseter and anterior temporal muscles was measured. Under experimentally altered occlusal conditions, the subjects performed clenching tasks at 50% of their maximal voluntary contraction level. Analysis of EMG responses revealed clenching on the unilateral occlusal support tended to cause a unilateral activity of the ipsilateral anterior temporalis. Analysis of the movement amplitude revealed a significant difference between the experimental occlusal conditions (P < 0.05). Clenching on unilateral occlusal stops caused a larger upward movement on the contralateral side. Bilateral first premolar clenching without molar support caused a larger upward movement of the mandible in the posterior region, whereas bilateral second molar clenching did not cause a significant upward movement. PMID- 11012856 TI - Cephalometric estimation of vertical dimension of occlusion. AB - The literature does not establish a single proven method for determining lower facial height, which is called the Vertical Dimension of Occlusion (VDO), and the concept of a vertical comfort range is generally accepted. This study aimed to test the statistical significance of correlations of mandibular shape versus lower facial height in occlusion, using cephalometric measurements. Correlations for 505 consecutive healthy adults were calculated between angles that estimate the lower facial height and angles that estimate the mandibular shape. The mandibular angle (gonial) showed a higher coefficient of correlation (r = 0.691) than the inferior gonial angle. The dispersion remained large, i.e. r2 = 0.478. Cephalometric measurements, despite theirs imperfections, could help the practitioner to understand what the best course of treatment would be in order to obtain a lower facial height in occlusion showing a skeletal harmony with the mandibular shape. PMID- 11012857 TI - Morphological interface between hybrid ionomers and dentin with and without smear layer removal. AB - To evaluate the micromorphological interface between dentin and several hybrid ionomer restoratives, a flat dentin surface was obtained on the occlusal surfaces of extracted human molar teeth after sectioning the enamel with an Isomet saw. Three poly-acid-modified composite resins, Compoglass, Dyract and F2000, and two resin-modified glass-ionomer cements, Fuji II LC and Photac-Fil were applied to the dentin surface. A second section, 2 mm apical from the first one, was made to produce a dentin segment containing the tested materials. Each disc was then split fractured along the dentin/material interface. For the poly-acid-modified composites, one half of the disc was stored in 6 mol/L HCl for 48 h to remove the dentin. The other was gently decalcified and deprotenized at the interface between the hybrid ionomer and the dentin. Both halves were then sputtered with gold and examined using SEM. For resin-modified glass-ionomer, samples were only evaluated at the interface. The three poly-acid-modified composite resins showed the formation of hybrid layers and resin tags at the interface to the dentin. Removal of the smear layer significantly improves hybridization of these materials. Also, Fuji II LC produced a hybrid layer while the Photac-Fil showed no evidence of hybridization. PMID- 11012858 TI - The temporomandibular opening index (TOI) in patients with closed lock and a control group with no temporomandibular disorders (TMD): an initial study. AB - Mandibular movement may be assessed by measuring maximum mouth opening. This is a linear measurement from the maxillary to the mandibular incisal edge and is affected by gender, age and ramus length. It cannot be used to classify patients. The temporomandibular opening index (TOI) appears to be independent of these variables and has been shown to differ for different categories of patients with temporomandibular disorders (TMD). This study examined the TOI in a group of 11 patients with closed lock and a control group of 11 individuals with no signs or symptoms of TMD. The group with closed lock had a significantly different TOI from that of the control group (P < 0.001) according to the Mann-Whitney U-test. This suggests that the TOI may have greater diagnostic value than linear mouth opening. PMID- 11012859 TI - A study on the mandibular movement of anterior openbite patients. AB - The purpose of this study was to compare the mandibular movements of anterior openbite patients using those of normal bite (angle class I) patients, to ascertain which components of mandibular movement are different in the two groups, and to use this information for occlusal treatment. The Saphon Visi trainer Model 3 and the Denar Pantronic were used to record mandibular movement and a Pantronic survey was performed using an arbitrary hinge axis, according to manufacturer's instructions. The subjects were 43 adults and included 28 subjects presenting with acceptable normal occlusion (angle class I) with no sign of TM dysfunction syndrome (TMD) and 15 subjects with anterior openbite with no anterior guidance. In the anterior openbite group, the average anterior and lateral condylar inclination, maximum opening and the distance between the intercuspal position with retruded contact position distance (anterior-posterior) were significantly lower than normal. The results suggest that in openbite patients the condyle inclination is flatter and the function of the TMJ is more restricted than in the mandibular movements of the normal group. It is hoped that these results will be useful for the correction of the anterior openbite condition. PMID- 11012861 TI - Simulated driving performance following prolonged wakefulness and alcohol consumption: separate and combined contributions to impairment. AB - The separate and combined effects of prolonged wakefulness and alcohol were compared on measures of subjective sleepiness, simulated driving performance and drivers' ability to judge impairment. Twenty-two males aged between 19 and 35 years were tested on four occasions. Subjects drove for 30 min on a simulated driving task under conditions determined by the factorial combination of 16 and 20 h of wakefulness and blood alcohol concentrations of 0.00 and 0.08%. The simulated driving session took place 30 min postingestion; subjects in the two alcohol conditions participated in a second 30-min driving session 90-min postingestion. Subjects made simultaneous ratings of their impairment while driving and retrospective ratings at the end of each test session. Subjective sleepiness measures were completed before and after each driving session. The combination of 20 h of prolonged wakefulness and alcohol produced significantly lower ratings of subjective sleepiness and driving performance that was worse, but not significantly so, than would be expected from the additive effects of each condition alone. Driving performance was always worse in the second driving session, during the elimination phase of alcohol metabolism, despite blood alcohol concentrations being lower than during the first driving session. There was a modest association between perceived and actual impairments in driving performance following prolonged wakefulness and alcohol. The findings suggest that the combination of prolonged wakefulness and alcohol consumption produced greater decrements in simulated driving performance than each condition alone and that drivers have only a modest ability to appreciate the magnitude of their impairment. PMID- 11012860 TI - Functional neuroimaging of normal human sleep by positron emission tomography. AB - Functional neuroimaging using positron emission tomography has recently yielded original data on the functional neuroanatomy of human sleep. This paper attempts to describe the possibilities and limitations of the technique and clarify its usefulness in sleep research. A short overview of the methods of acquisition and statistical analysis (statistical parametric mapping, SPM) is presented before the results of PET sleep studies are reviewed. The discussion attempts to integrate the functional neuroimaging data into the body of knowledge already acquired on sleep in animals and humans using various other techniques (intracellular recordings, in situ neurophysiology, lesional and pharmacological trials, scalp EEG recordings, behavioural or psychological description). The published PET data describe a very reproducible functional neuroanatomy in sleep. The core characteristics of this 'canonical' sleep may be summarized as follows. In slow-wave sleep, most deactivated areas are located in the dorsal pons and mesencephalon, cerebellum, thalami, basal ganglia, basal forebrain/hypothalamus, prefrontal cortex, anterior cingulate cortex, precuneus and in the mesial aspect of the temporal lobe. During rapid-eye movement sleep, significant activations were found in the pontine tegmentum, thalamic nuclei, limbic areas (amygdaloid complexes, hippocampal formation, anterior cingulate cortex) and in the posterior cortices (temporo-occipital areas). In contrast, the dorso-lateral prefrontal cortex, parietal cortex, as well as the posterior cingulate cortex and precuneus, were the least active brain regions. These preliminary studies open up a whole field in sleep research. More detailed explorations of sleep in humans are now accessible to experimental challenges using PET and other neuroimaging techniques. These new methods will contribute to a better understanding of sleep functions. PMID- 11012862 TI - Diurnal variations in the waking EEG: comparisons with sleep latencies and subjective alertness. AB - Daytime measures of sleep latency and subjective alertness do not correlate with one another, suggesting that they assess different aspects of alertness. In addition, their typical diurnal variations show very different time courses. Quantitative analysis of the waking electroencephalogram (EEG) has been proposed as an objective measure of alertness, but it is not clear how it compares with other measures. In this study, the waking EEG was measured in the daytime to determine the presence of diurnal variations in the activity of standard frequency bands and to compare these variations with the temporal patterns typical of sleep propensity and subjective alertness. Alertness was evaluated in four men and 12 women, aged 19-33 y. Assessments were conducted every 2 h, from 10.00 to 24.00, in the following order: a visual analogue scale of alertness, a waking EEG recording and a sleep latency test. The waking EEG was recorded with eyes open. For each recording session, 32-60 s of artefact-free signals were selected from the C3/A2 derivation, then subjected to amplitude spectral analysis. Four EEG frequency bands showed significant diurnal variations: delta, theta, sigma and beta1. None of these variations showed a significant correlation with the temporal patterns of sleep latencies or subjective alertness. At the individual level, however, theta band activity increased when subjective alertness decreased, suggesting that the theta band can be used to monitor variations in alertness in a given individual, even at the moderate levels of sleepiness experienced during the daytime. PMID- 11012863 TI - Gender-related sleep differences in neonates in thermoneutral and cool environments. AB - Although thermoregulation and sleep exhibit gender differences in adults, the question is still debated in neonates. The aim of this study was to examine the relationship between gender-related sleep differences and cool defence mechanisms in neonates. Sleep and thermoregulation were recorded in healthy preterm neonates (21 boys and 17 girls, 37 +/- 2 weeks post-conceptional age) exposed to thermoneutral and cool conditions. Sleep was analysed for continuity and structure. Although the cool exposure did not strongly impair body homeothermia, sleep was altered but without any significant gender difference. However, when data recorded under each of the thermal conditions were pooled, some gender differences emerged: boys slept less, with more wakefulness after sleep onset, more active sleep and less quiet sleep than girls. In contrast to sleep architecture, most of the sleep continuity parameters exhibited greater variability in boys than in girls. This variability may bias the statistical analyses and probably explains the varying conclusions reported in the literature regarding gender-specific sleep-related differences. PMID- 11012864 TI - Tactile arousal threshold of sleeping king penguins in a breeding colony. AB - The tactile arousal threshold of sleeping birds has not been investigated to date. In this study, the characteristics of this threshold were assessed by stimulating either the upper back or a foot of two groups (one cutaneous site per group) of 60 sleeping king penguins (Aptenodytes patagonica) in the breeding colony of Baie du Marin (Crozet Archipelago). Increasing weights were put onto one of the feet or the upper back of individuals that had been sleeping for more than 5 min until they showed behavioural signs of arousal (head raising). The weight applied to the upper back that was needed to awaken a sleeper (837 +/- 73 g) was 20 times greater than that applied to a foot (38 +/- 6 g). In terms of pressure, the difference remained five times higher for the back (209 +/- 18 g/cm(2)) than the foot (40 g +/- 7 g/cm(2)). Because the king penguin incubates its single egg and rears its young chick on its feet, the low threshold measured at this level could be viewed as an adaptation against progeny predation. Sleepers are frequently bumped by conspecifics walking through the colony. The increased arousal threshold associated with tactile stimulation of the back may help to preserve sleep continuity under these conditions. PMID- 11012865 TI - Rest and activity states in a gray whale. AB - The behaviour of a female gray whale (Eschrichtius robustus) that had been rescued 14 months previously was recorded continuously on a video-recorder for 9 days at 'Sea World' in San Diego. On average, during the first six recording days, active wakefulness accounted for 37.9 +/- 1.7% of each 24 h; transitional stage for 17.4 +/- 1.4% and rest for 41.2 +/- 1.7%. In the rest stage the whale was lying on the bottom of the pool (13.2 +/- 1.7%) or hanging on the surface (28. 0 +/- 1.7%). During the rest stage, it was immobile most of the time and moved only for respiration. In the rest stage both eyes could be open, one eye could be open while the other was closed or, more rarely, both eyes could be closed. Characteristic jerks of the head, neck and sometimes of the whole body were observed in the whale during the rest stage. Most jerks were single and only 10% of all jerks were serial (occurring within 10 s of a prior jerk). Eyelid movements accompanied 40% of jerks. In two episodes, intense jerks followed each other continuously for 3 and 4 s and were accompanied by eyelid movements. These jerks resembled the twitches characteristic of paradoxical sleep in terrestrial mammals. During these episodes the whale was falling slowly onto its side and subsequently started to swim in the pool. PMID- 11012866 TI - Does caffeine confound relationships among adrenergic tone, blood pressure and sleep apnoea? AB - The purpose of this study was to determine whether caffeine consumption confounds the relationship among adrenergic tone, as measured by urinary norepinephrine (NE), blood pressure (BP) and obstructive sleep apnoea (OSA). Data were analysed using correlation and regression analysis, analysis of covariance and t-tests. Subjects included normotensives and hypertensives with and without OSA: 38 men, 23 women, aged 30-60 y; 100-150% of ideal body weight; without other major illness. Patients were studied using polysomnography, caffeine consumption was assessed, 24-h urinary NE levels were examined and ambulatory BP was recorded. Patients with OSA (N=27) reported significantly greater caffeine consumption than those without OSA (N=34) (295 vs. 103 mg, P=0.010), but caffeine was not significantly correlated with their ambulatory BP. In contrast, NE excretion correlated with caffeine consumption (r=0.24, P=0.041), apnoea severity (r=0.65, P < 0.001) and BP (r=0.34, P < 0.005). Significant OSA-NE and BP-NE relationships remained even after controlling for caffeine consumption. Patients with OSA consumed nearly three times the amount of caffeine as patients without OSA. While caffeine partially explains the increased adrenergic tone in patients with OSA and the relationship between BP and NE, it does not appear to contribute significantly to the relationship between OSA and elevated BP. PMID- 11012867 TI - EEG arousals and awakenings in relation with periodic leg movements during sleep. AB - It is known that periodic leg movements are frequently accompanied by full awakenings or by signs of EEG arousals. The time relationship of these EEG arousals with leg movements varies from patient to patient. They may precede or follow leg movements or occur simultaneously. It is not clear whether these arousals trigger leg movements or, alternatively, whether both EEG arousals and leg movements are separate expressions of a common pathophysiological mechanism. We investigated the temporal relationship of five EEG arousals, such as alpha activity, K-complexes, spindles, K-alpha, K-spindle activities and awakenings, with leg movements in 10 periodic leg movement patients. These EEG arousals were considered to be associated with leg movements if they occurred 10 s before/after or simultaneously with the onset of right or left tibialis muscle EMG potentials. It was found that 49.19% of EEG arousals occurred before leg movements, 30.61% occurred simultaneously and 23.18% occurred just after leg movements. The number of EEG arousals was significantly higher in the 10 s preceding leg movement than simultaneously or in the 10 s following. Alpha activity was the phenomenon associated most frequently with leg movements, irrespective of its temporal organization and was significantly higher during the 10 s preceding movement. Spindle and K-spindle activities were significantly higher before leg movement, whereas K-complex activity was significantly more frequent during leg movements. The number of awakenings was significantly higher after leg movements than simultaneously. These results indicated that leg movements are not primary, but rather are a phenomenon associated with an underlying arousal disorder. PMID- 11012868 TI - Computerized endopharyngeal myotonometry (CEM): a new method to evaluate the tissue tone of the soft palate in patients with obstructive sleep apnoea syndrome. AB - This study compared the tissue tone of the soft palate in nonsnoring subjects and patients with obstructive sleep apnoea syndrome (OSAS) during wakefulness. Here, tissue tone means the biomechanical property of the tissue which can be characterized by two main parameters: stiffness and elasticity. Tissue tone includes both structural and neural components. A new method to evaluate the tissue tone of the soft palate was used - computerized endopharyngeal myotonometry (CEM). This method records and analyses the response of the soft palate tissues to a brief mechanical impact. The method enabled us to evaluate the most important parameters of tissue tone: stiffness, which is expressed as a frequency; and elasticity, expressed as a logarithmic decrement of the damped oscillation. First, a self-reported questionnaire was completed about the medical history of the subjects. Subjects then underwent a physical examination of the oropharynx and polysomnography with overnight pulse oximetry. The results of the CEM method indicated that patients with OSAS show an increased stiffness of the soft palate tissues (20.3, SD 4.7 Hz) compared with nonsnoring subjects (12.2, SD 1.8 Hz). In patients with sleep apnoea, elasticity is not increased in a similar way to stiffness. Thus, the disproportion between tissue stiffness and elasticity of the soft palate is a measure of the pathological changes in patients with sleep apnoea. PMID- 11012869 TI - Increased sodium-proton antiporter activity in patients with obstructive sleep apnoea. AB - Cytosolic pH (pH(i)) and the activity of the sodium-proton antiporter (Na(+)/H(+) antiporter) were measured in lymphocytes from 22 patients with obstructive sleep apnoea and from 24 age-matched healthy subjects (Controls). The cellular Na(+)/H(+) antiporter was measured spectrophotometrically using a pH-sensitive fluorescent dye after intracellular acidification using sodium propionate. Resting pHi was similar in lymphocytes from patients with obstructive sleep apnoea and from controls (7.36 +/- 0.20, n=22; vs. 7.35 +/- 0.19, n=24; mean +/- SD). The Na(+)/H(+) antiporter activity was significantly higher in patients with obstructive sleep apnoea than in controls (11.87 +/- 3.26 x 10(-3) pH(i)/s vs. 4.38 +/- 1.40 x 10(-3) pH(i)/s; P < 0. 0001). The apparent affinity of the Na+/H+ antiporter was not significantly different between the groups (6.90 +/- 0.23 vs. 6.87 +/- 0.20). In patients with obstructive sleep apnoea the activity of the Na(+)/H(+) antiporter remained stable during the night. The activity of the Na(+)/H(+) antiporter was 13.49 +/- 4.80 x 10(-3) pH(i)/s at 20.00 and 13.26 +/- 6.13 x 10(-3) pH(i)/s at 02.00. From the present results it is concluded that an increased cellular Na(+)/H(+) antiporter activity may be a genetic marker for patients who are predisposed to obstructive sleep apnoea. PMID- 11012870 TI - Excessive daytime sleepiness in patients suffering from different levels of obstructive sleep apnoea syndrome. AB - Excessive daytime sleepiness (EDS) is a frequent symptom of patients with obstructive sleep apnoea (OSA). EDS is a high-risk factor for accidents at work and on the road. Thirty untreated patients with different levels of severity of OSA were studied concerning night sleep and EDS. The criterion for severity was the respiratory disturbance index (RDI): 15 patients were classified as 'moderately' apnoeic (RDI < 40), 15 as 'severely' apnoeic (RDI > 40). Following night-time polysomnography, objective and subjective aspects of EDS were studied. To assess objective EDS the Maintenance of Wakefulness Test (MWT) and a computer based vigilance performance test were used. Subjective EDS was determined using the Stanford Sleepiness Scale (SSS), the Epworth Sleepiness Scale (ESS) and the Visual Analogue Scales for Performance (VAS-P) and Tiredness (VAS-T). Well-being was assessed using the Scale of Well-Being by von Zerssen (Bf-S/Bf-S'). Severe apnoea patients spent more time in stage 1 and less in slow-wave sleep. MWT latencies tended to be shorter in the severe apnoea group. Vigilance testing revealed no group differences. Patients with moderate apnoea described themselves as more impaired in all subjective scales, but only SSS scores reached statistical significance. Our results suggest that there is no simple correlation between polysomnographic and respiratory sleep variables at night on the one hand, and the extent of EDS on the other hand. Furthermore, subjective and objective evaluation of EDS does not yield the same results. New approaches which allow a more detailed analysis of night sleep and daytime function are required to identify high-risked patients. PMID- 11012871 TI - Quality of life assessment of treatment with dental appliance or UPPP in patients with mild to moderate obstructive sleep apnoea. A prospective randomized 1-year follow-up study. AB - The objectives of this study were: to evaluate the change in the three quality of life (QOL) dimensions of vitality, contentment and sleep before intervention and 1 year after treatment with a dental appliance or uvulopalatopharyngoplasty (UPPP); to compare the effect of treatment between these two treatment groups on these three dimensions; and to determine the relation between the QOL scores and somnographic values. Ninety-five patients with mild to moderate obstructive sleep apnoea (OSA) (AI > 5) were randomly allocated to either a dental appliance or UPPP treatment group. Seven patients withdrew after randomization but before treatment, leaving 88 patients eligible for treatment. The patients were examined using somnography and administered the Minor Symptoms Evaluation-Profile (MSE-P), a QOL questionnaire, before and 1 year after intervention. Thirty-seven patients in the dental appliance group and 43 in the UPPP group completed the 1-year follow-up. The mean values for the three dimensions vitality, contentment and sleep improved significantly 1 year after intervention in the dental appliance and UPPP groups. No difference in the QOL scores at baseline was noted between the groups. One year after intervention the UPPP group showed significantly more contentment than the dental appliance group. In contrast, vitality and sleep dimensions did not differ between the two treatment groups. No significant correlations were observed between the QOL scores and somnographic values. In conclusion, quality of life improved significantly in the dental appliance and UPPP groups 1 year after intervention. However, the dental appliance group showed a lower level of contentment than the UPPP group, even though the somnographic values were superior in the former group. PMID- 11012872 TI - Night-time sleep and daytime sleepiness in narcolepsy. AB - This report describes night-time sleep and daytime sleepiness in a large (N=530) sample of patients meeting the International Classification of Sleep Disorders criteria for diagnosis of narcolepsy. Sleep data were obtained from polysomnographic recordings on two consecutive nights. Sleepiness was assessed using the Multiple Sleep Latency Test, the Maintenance of Wakefulness Test and the Epworth Sleepiness Scale. Analysis revealed that sleep was mild to moderately disturbed on both recording nights. A first-night effect was suggested by decreased REM latency and increased percentage REM and slow-wave sleep on the second night. Sleepiness and sleep disturbance varied across patient subgroups created based on patient ethnicity and on the presence/absence of cataplexy, sleep apnoea, and periodic limb movements. Covariation of sleep and sleepiness measures across patients was significant but weak. Strong association was found between subgroup means of sleep and sleep disturbance measures. The findings reported here show that sleepiness and sleep disturbance vary across patient subgroups and that sleep disturbance is related to, although unable to account, for the pathological sleepiness of narcolepsy. PMID- 11012873 TI - Treatment of advanced renal failure: low-protein diets or timely initiation of dialysis? AB - Until 1996, no guidelines existed for the initiation of dialysis in patients with progressive renal failure. The publication of the National Kidney Foundation Dialysis Outcome Quality Initiative guidelines has generated a debate on the management of advanced renal failure and the role of low-protein diets (LPDs). We performed a review of the literature to identify articles on the initiation of dialysis and LPDs, particularly those since 1996. Delayed referral of patients is widespread in both the United States and Europe, and almost 25% of patients are started on dialysis at a glomerular filtration rate (GFR) of <5 mL/min/1.73 m2. There is a high prevalence of malnutrition at the time of first dialysis, which progressively improves upon initiation of dialysis. There is no evidence regarding the efficacy or safety of LPDs in nondiabetic patients younger than 70 years old [approximately 40% of U.S. incident end-stage renal disease (ESRD) patients] and in diabetics with GFR <25 mL/min/1.73 m2 (>40% of incident U.S. ESRD). In nondiabetics who are younger than 70 years old, adherence to LPD for four to five years can be estimated to result in a delay in dialysis by 6 to 11 months. However, suboptimal energy intake is widespread in advanced renal failure, which declines further upon institution of LPD. Even nutritionally sound patients develop subclinical nutritional decline despite intense counseling. There are no data on the efficacy or safety of LPD in subgroups that constitute approximately 80% of incident ESRD patients. Concerns still exist regarding their nutritional safety in the remainder. Initiation of dialysis results in improved nutritional status and should be considered in a timely fashion. PMID- 11012874 TI - Connective tissue growth factor: potential role in glomerulosclerosis and tubulointerstitial fibrosis. AB - Transforming growth factor beta (TGF-beta) is a pivotal driver of glomerulosclerosis and tubulointerstitial fibrosis in renal diseases. Because TGF beta also plays important anti-inflammatory and antiproliferative roles in mammalian systems, there has been a recent drive to elucidate downstream mediators of TGF-beta's pro-fibrotic effects with the ultimate goal of developing new anti-fibrotic strategies for treatment of chronic diseases. Connective tissue growth factor (CTGF) belongs to the CCN family of immediate early response genes. Several lines of evidence suggest that CTGF is an important pro-fibrotic molecule in renal disease and that CTGF contributes to TGF-beta bioactivity in this setting. CTGF expression is increased in the glomeruli and tubulointerstium in a variety of renal disease in association with scarring and sclerosis of renal parenchyma. In model systems in vitro, mesangial cell CTGF expression is induced by high extracellular glucose, cyclic mechanical strain and TGF-beta. Recombinant human CTGF augments the production of fibronectin and type IV collagen by mesangial cells and the effects of high glucose on mesangial cell CTGF expression and matrix production are attenuated, in part, by anti-TGF-beta antibody. In aggregate, these observations identify CTGF as an attractive therapeutic target in fibrotic renal diseases. PMID- 11012875 TI - Thirteen novel mutations of the replicated region of PKD1 in an Asian population. AB - BACKGROUND: Mutations of PKD1 are thought to account for approximately 85% of all mutations in autosomal dominant polycystic kidney disease (ADPKD). The search for PKD1 mutations has been hindered by both its large size and complicated genomic structure. To date, few mutations that affect the replicated segment of PKD1 have been described, and virtually all have been reported in Caucasian patients. METHODS: In the present study, we have used a long-range polymerase chain reaction (PCR)-based strategy previously developed by our laboratory to analyze exons in the replicated region of PKD1 in a population of 41 unrelated Thai and 6 unrelated Korean families with ADPKD. We have amplified approximately 3.5 and approximately 5 kb PKD1 gene-specific fragments (5'MR and 5'LR) containing exons 13 to 15 and 15 to 21 and performed single-stand conformation analysis (SSCA) on nested PCR products. RESULTS: Nine novel pathogenic mutations were detected, including six nonsense and three frameshift mutations. One of the deletions was shown to be a de novo mutation. Four potentially pathogenic variants, including one 3 bp insertion and three missense mutations, were also discovered. Two of the nonconservative amino acid substitutions were predicted to disrupt the three dimensional structure of the PKD repeats. In addition, six polymorphisms, including two missense and four silent nucleotide substitutions, were identified. Approximately 25% of both the pathogenic and normal variants were found to be present in at least one of the homologous loci. CONCLUSION: To our knowledge, this is the first report of mutation analysis of the replicated region of PKD1 in a non-Caucasian population. The methods used in this study are widely applicable and can be used to characterize PKD1 in a number of ethnic groups using DNA samples prepared using standard techniques. Our data suggest that gene conversion may play a significant role in producing variability of the PKD1 sequence in this population. The identification of additional mutations will help guide the study of polycystin-1 and better help us to understand the pathophysiology of this common disease. PMID- 11012876 TI - Structural analysis of the 11beta-hydroxysteroid dehydrogenase type 2 gene in end stage renal disease. AB - BACKGROUND: Mutations in the 11beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2) gene cause a rare form of low-renin hypertension leading to end stage renal disease (ESRD) in some affected subjects. To date, no search for mutations in the HSD11B2 gene was performed in a large population to obtain an estimate its prevalence. METHODS: The HSD11B2 gene was analyzed in 587 subjects, including 260 ESRD patients (either dialysis or transplanted) for mutations in the exons 2 through 5 and corresponding intronic regions by polymerase chain reaction (PCR) using appropriate overlapping primers, gel analysis by single strand conformational polymorphism (SSCP), and sequencing of identified migration variants. RESULTS: The prevalence of single-nucleotide polymorphisms (SNPs) in ESRD patients and controls was 26%. The following genetic variants were found among all subjects investigated: exon 2 T442G (Leu148/Val, N = 70) and C470A (Thr156/Thr, N = 67), exon 3 G534A (Glu178/Glu, N = 69), and exon 5 C1274T (Asp388/Asp, N = 2). Four SNPs were identified in intron 4 only. In the control population, the prevalence of the variants Leu148 and Thr156 was 14% each. Glu178 was 11%, while no variants were found in exon 5. In ESRD patients, the prevalence of the variant Leu148 was 9%, and Thr156 was 8%. Glu178 was 13%, while the Asp388 variant was 0.7%. In patients with a short duration between the time of diagnosis of the renal disease and the onset of ESRD, the prevalence of the Leu148 and Glu178 variants was higher than in subjects with slowly progressing renal disease. The 11betaHSD2 activity of all of these SNPs is predictably unaltered. CONCLUSIONS: There is a high prevalence of SNPs of the HSD11B2 gene, without causing exonic mutations generating a 11betaHSD2 enzyme with altered activity. Based on statistical analyses, the frequency of homozygosity for mutated alleles of the HSD11B2 gene can be derived as <1/250,000 when a Caucasian population is considered. PMID- 11012877 TI - Cerivastatin prevents angiotensin II-induced renal injury independent of blood pressure- and cholesterol-lowering effects. AB - BACKGROUND: Statins are effective in prevention of end-organ damage; however, the benefits cannot be fully explained on the basis of cholesterol reduction. We used an angiotensin II (Ang II)-dependent model to test the hypothesis that cerivastatin prevents leukocyte adhesion and infiltration, induction of inducible nitric oxide synthase (iNOS), and ameliorates end-organ damage. METHODS: We analyzed intracellular targets, such as mitogen-activated protein kinase and transcription factor (nuclear factor-kappaB and activator protein-1) activation. We used immunohistochemistry, immunocytochemistry, electrophoretic mobility shift assays, and enzyme-linked immunosorbent assay techniques. We treated rats transgenic for human renin and angiotensinogen (dTGR) chronically from week 4 to 7 with cerivastatin (0.5 mg/kg by gavage). RESULTS: Untreated dTGR developed hypertension, cardiac hypertrophy, and renal damage, with a 100-fold increased albuminuria and focal cortical necrosis. dTGR mortality at the age of seven weeks was 45%. Immunohistochemistry showed increased iNOS expression in the endothelium and media of small vessels, infiltrating cells, afferent arterioles, and glomeruli of dTGR, which was greater in cortex than medulla. Phosphorylated extracellular signal regulated kinase (p-ERK) was increased in dTGR; nuclear factor-kappaB and activator protein-1 were both activated. Cerivastatin decreased systolic blood pressure compared with untreated dTGR (147 +/- 14 vs. 201 +/- 6 mm Hg, P < 0.001). Albuminuria was reduced by 60% (P = 0.001), and creatinine was lowered (0.45 +/- 0.01 vs. 0.68 +/- 0.05 mg/dL, P = 0. 003); however, cholesterol was not reduced. Intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression was diminished, while neutrophil and monocyte infiltration in the kidney was markedly reduced. ERK phosphorylation and transcription factor activation were reduced. In addition, in vitro incubation of vascular smooth muscle cells with cerivastatin (0.5 micromol/L) almost completely prevented the Ang II-induced ERK phosphorylation. CONCLUSION: Cerivastatin reduced inflammation, cell proliferation, and iNOS induction, which led to a reduction in cellular damage. Our findings suggest that 3-hydroxy-3-methylglutaryl coenzyme (HMG-CoA) reductase inhibition ameliorates Ang II-induced end-organ damage. We suggest that these effects were independent of cholesterol. PMID- 11012878 TI - Stretch activation of jun N-terminal kinase/stress-activated protein kinase in mesangial cells. AB - BACKGROUND: Mesangial cells (MCs) grown on extracellular matrix (ECM)-coated plates and exposed to cyclic stretch/relaxation proliferate and produce ECM protein, suggesting that this may be a useful in vitro model for MC behavior in response to increased physical forces. The induction of c-fos in response to MC stretch has been shown. Stimuli that lead to c-fos induction pass through mitogen activated protein (MAP) kinase pathways. We have seen early activation of jun N terminal kinase/stress-activated protein kinase (SAPK/JNK) in MCs exposed to cyclic stretch. Accordingly, we studied SAPK/JNK activation in stretched MCs and the downstream consequences of this signaling. METHODS: MCs (passages 5 to 10) cultured on type 1 collagen-coated, flexible-bottom plates were exposed to 2 to 60 minutes of cyclic strain (60 cycles per minute) by generation of vacuums of 10 to -27 kPa, inducing approximately 16 to 28% maximum elongation in the diameter of the surfaces. Control MCs were grown on coated rigid bottom plates. Protein levels (by Western blot) and activity assays for SAPK/JNK were performed under these conditions. We observed marked activation at -18 kPa and above and at two minutes, and then we studied activation mechanisms under these conditions. Nuclear protein binding to activator protein-1 (AP-1) consensus sequences was also examined. The role of calcium was studied with EGTA and BAPTA-AM to chelate extra- and intracellular calcium, respectively. Protein kinase C (PKC) was down regulated by incubation with phorbol ester (PMA) for 24 hours prior to stretch. In unstretched MCs, A23187 was used as a calcium ionophore, and PKC was up regulated with PMA application for 30 minutes to determine the effects on SAPK/JNK. Nuclear protein binding to AP-1 was also determined under these conditions. The effects of stretch, acute PMA, and A23187 on fibronectin mRNA levels were studied using reverse transcriptase-polymerase chain reaction (RT PCR). RESULTS: Cyclic strain/relaxation led to increased SAPK/JNK activity only at two minutes and -18 kPa and above. The activation of SAPK/JNK was dependent on intracellular calcium, with BAPTA-AM almost completely abrogating the response to stretch. EGTA was without effect. Down-regulation of PKC also led to a diminution of activity. In static cells, the calcium ionophore A23187 increased SAPK/JNK activity, and this was potentiated by acute PMA. Stretch, acute PMA, and A23187 all increased nuclear protein binding to AP-1 consensus sequences. mRNA levels for fibronectin were increased by stretch in MCs and by PMA and A23187 in static MCs. No change was observed in the amount of SAPK/JNK protein present in stretched MCs by Western blot. CONCLUSIONS: Stretch leads to early activation of SAPK/JNK in MCs. This is dependent on intracellular calcium and PKC and can be replicated by activation of these stimuli in static MCs. A downstream induction of nuclear protein binding to AP-1 consensus sequences was seen in a pattern that was completely concordant with the SAPK/JNK induction. PMID- 11012879 TI - Down-regulation of human osteoblast PTH/PTHrP receptor mRNA in end-stage renal failure. AB - BACKGROUND: Resistance to the action of parathyroid hormone (PTH) has been demonstrated in end-stage renal failure and is considered to be important in the pathogenesis of secondary hyperparathyroidism. The mechanism of resistance is unknown. However, altered regulation of cellular PTH/PTH-related protein (PTH/PTHrP) receptor (PTH1R) has been assumed to be important. METHODS: We have used in situ hybridization to examine PTH1R mRNA expression by osteoblasts in human bone and have compared the expression in high- and low-turnover renal bone disease, high-turnover nonrenal bone disease (healing fracture callus and Pagetic bone), and normal bone. Bone biopsies were formalin fixed, ethylenediaminetetraacetic acid decalcified, and paraffin wax embedded. A 1.8 kb PTH1R cDNA probe, labeled with 35S, was used, and the hybridization signal was revealed by autoradiography. The density of signal over osteoblasts was quantitated using a semiautomated Leica image analysis software package. RESULTS: The mean density of PTH1R mRNA signal over osteoblasts in renal high-turnover bone was only 36% of that found in nonrenal high-turnover bone (P < 0.05) and 51% of that found in normal bone (P < 0.05). Osteoblast PTH1R mRNA signal in adynamic bone from individuals with diabetes mellitus was 28% of normal bone (P < 0.05) and 54% of that found in renal high-turnover bone (P < 0.05). CONCLUSIONS: These results demonstrate a down-regulation of osteoblast PTH1R mRNA in end-stage renal failure in comparison to normal and high-turnover bone from otherwise healthy individuals, and provide an insight into the mechanisms of "skeletal resistance" to the actions of PTH. PMID- 11012880 TI - Functional modulation of the mineralocorticoid receptor by cis diamminedichloroplatinum (II). AB - BACKGROUND: Renal salt wasting and hypotension are some of the frequent complications in patients treated with cis-diamminedichloroplatinum (II) (cDDP), and it is suggested that cDDP produces an abnormality in the renin-angiotensin system. However, not only the underlying mechanism but also prophylactic treatment of this cDDP toxicity remains unknown. In the present study, we investigated the molecular mechanism of this cDDP-induced disturbance of renal sodium handling with focusing on the effect of cDDP on mineralocorticoid receptor (MR) function. METHODS: The effect of cDDP was studied on nuclear translocation, DNA binding activity, and transactivation function of the MR. RESULTS: In a transient transfection assay, cDDP suppressed MR-dependent reporter gene expression. This cDDP-mediated repression of MR function, at least in part, is suggested to be due to the generation of reactive oxygen species and a subsequent decrease in ligand-dependent nuclear translocation and suppression of the interaction with DNA of the MR. This redox-dependent repression of MR function both in vitro and in vivo was reversed by treatment with reducing reagents. Moreover, cDDP, most possibly via formation of DNA adducts, inhibited MR-DNA interaction in a redox-independent fashion. CONCLUSIONS: MR function is impaired by cDDP at multiple levels, via redox-dependent and -independent mechanisms. PMID- 11012881 TI - Nephrin in experimental glomerular disease. AB - BACKGROUND: The recently identified gene NPHS1 with its mutations causing congenital nephrotic syndrome of the Finnish type (CNF) is highly promising in providing new understanding of pathophysiology of proteinuria. Earlier we cloned a rat NPHS1 homologue, as well as characterized and raised antibodies to the respective protein product nephrin. METHODS: Changes in the expression levels of nephrin-specific mRNA in commonly used experimental models of proteinuria were examined using semiquantitative reverse transcription-polymerase chain reaction, immunofluorescence, and immunoelectron microscopy (IEM) of nephrin. RESULTS: Notably, a 40% down-regulation of the nephrin-specific mRNA of cortical kidney was seen already at day 3 after induction of the puromycin aminonucleoside nephrosis (PAN), while no major elevation of urinary protein secretion was seen at this stage. A further decrease of 80% of nephrin message was seen at the peak of proteinuria at day 10. A similar decrease of up to 70% from the basal levels was seen in mercuric chloride-treated rats. Changes in the protein expression paralleled those of the mRNA in indirect immunofluorescence. Interestingly, a remarkable plasmalemmal dislocation from the normal expression site at the interpodocyte filtration slits could be observed in IEM. CONCLUSIONS: Nephrin appears to be an important causative molecule of proteinuria and shows a remarkable redistribution from the filtration slits to the podocyte plasma membrane, especially in PAN. PMID- 11012882 TI - Osteopontin expression in progressive renal injury in remnant kidney: role of angiotensin II. AB - BACKGROUND: Osteopontin (OPN) is a macrophage chemotactic and adhesion molecule and has been shown to play a role in glomerular and tubulointerstitial injury in several kidney disease models. METHODS: The present study examined whether OPN expression is involved in the progression of renal disease following subtotal (5/6) nephrectomy (STNx) in rats and whether angiotensin II (Ang II) mediates the up-regulation of renal OPN expression and macrophage accumulation in this model by administering valsartan, an Ang II type I (AT1) receptor antagonist, or ramipril, an angiotensin-converting enzyme (ACE) inhibitor. RESULTS: In normal and sham-operated rat kidneys, OPN was expressed in a few tubules (<5%) and was absent in glomeruli. Following STNx (weeks 2 to 16), there was substantial up regulation of OPN mRNA and protein expression in glomeruli [2 to 12 cells/glomerular cross section (gcs)] and tubular epithelial cells (20 to 75% OPN+). The up-regulation of OPN expression was associated with macrophage accumulation within the kidney, severe proteinuria, loss of renal function, and severe histologic damage, including tubulitis and tubulointerstitial fibrosis (all P < 0.001). Treatment with either valsartan or ramipril completely abrogated the up-regulation of OPN mRNA and protein expression in glomeruli and tubules. The reduction in OPN expression was associated with a significant inhibition of macrophage accumulation and progressive renal injury (P < 0.001). CONCLUSION: An up-regulation of OPN expression may play a role in progressive renal injury following STNx. Inhibition of OPN expression may be one of the mechanisms by which Ang II blockade attenuated renal injury after renal ablation. PMID- 11012883 TI - Mechanism of chronic obstructive uropathy: increased expression of apoptosis promoting molecules. AB - BACKGROUND: We have demonstrated that renal tubular and interstitial cells undergo pronounced apoptosis during the course of chronic obstructive uropathy (COU). Apoptosis is a complex cellular process consisting of multiple steps, each of which is mediated by families of related molecules. These families may include receptor/ligand molecules such as Fas, Fas ligand, tumor necrosis factor receptor 1 (TNFR-1), and TNF-related apoptosis inducing ligand (TRAIL); signal transduction adapter molecules such as Fas-associated death domain (FADD), TNFR-1 associated death domain (TRADD), receptor-interacting protein (RIP), Fas associated factor (FAF), and Fas-associated phosphatase (FAP); or effector molecules such as caspases. However, the mechanism of tubular cell apoptosis, as well as the pathogenetic relevance of these apoptosis-related molecules in COU, remains poorly understood. METHODS: Kidneys were harvested from sham-operated control mice and mice with COU created by left ureter ligation sacrificed in groups of three at days 4, 15, 30, and 45. To detect apoptotic tubular and interstitial cells, in situ end labeling of fragmented DNA was performed. To detect the expression of apoptosis-related molecules, ribonuclease protection assay was used with specific antisense RNA probes for Fas, Fas ligand, TNFR-1, TRAIL, FADD, TRADD, RIP, FAF, FAP, and caspase-8. Immunostaining for Fas, Fas ligand, TRAIL, TRADD, RIP, and caspase-8 was also performed. To assess the role of these molecules in COU-associated renal cell apoptosis, the frequencies of apoptotic tubular and interstitial cells were separately quantitated for each experimental time point, and their patterns of variation were correlated with those of apoptosis-related molecules. RESULTS: The obstructed kidneys displayed increased apoptosis of both tubular and interstitial cells. Tubular cell apoptosis appeared at day 4 after ureter ligation, peaked (fivefold of control) at day 15, and decreased gradually until the end of the experiment. In contrast, interstitial cell apoptosis sustained a progressive increase throughout the experiment. Apoptosis was minimal at all experimental time points for control and contralateral kidneys. Compared with control and contralateral kidneys, the ligated kidneys displayed a dynamic expression of mRNAs for many apoptosis related molecules, which included an up to threefold increase for Fas, Fas ligand, TNF-R1, TRAIL, TRADD, RIP, and caspase-8, and an up to twofold increase for FADD and FAP, but there was little change for FAF. These mRNAs increased between days 4 and 15, decreased until day 30, but then increased again until day 45. The rise and fall of mRNAs between days 4 and 30 paralleled a similar fluctuation in tubular cell apoptosis in that period. The subsequent increase of mRNAs was correlated with a continuous rise of interstitial cell apoptosis. We demonstrated a positive immunostaining for Fas and Fas ligand in the tubular cells at early time points as well as in interstitial inflammatory cells at later time points. Although increased expression of TRAIL, TRADD, RIP, and caspase-8 was noted in tubular cells, there was no staining for these molecules in interstitial cells. CONCLUSION: The current study documents a dynamic expression of several molecules that are known to mediate the most crucial steps of apoptosis. It implicates these molecules in COU-associated renal cell apoptosis and in the pathogenesis of this condition. It also lays the foundation for interventional studies, including genetic engineering, to evaluate the molecular control of apoptosis associated with COU. PMID- 11012884 TI - Up-regulation of monocyte chemoattractant protein-1 in tubulointerstitial lesions of human diabetic nephropathy. AB - BACKGROUND: We previously described that monocyte chemoattractant protein-1 (MCP 1) plays an important role in progressive glomerular and interstitial damage in inflammatory renal diseases. However, the expression of MCP-1 in diabetic nephropathy remains to be investigated. METHODS: We examined whether locally expressed MCP-1 participates in human diabetic nephropathy via recruiting and activating monocytes/macrophages (Mphi). Urinary and serum MCP-1 levels were measured by enzyme-linked immunosorbent assay in 45 patients with diabetic nephropathy. The presence of MCP-1 in diseased kidneys was determined by immunohistochemical and in situ hybridization analyses. RESULTS: Urinary MCP-1 levels were significantly elevated in patients with diabetic nephrotic syndrome and advanced tubulointerstitial lesions. Moreover, urinary levels of MCP-1 were well correlated with the number of CD68-positive infiltrating cells in the interstitium. In contrast, serum MCP-1 levels remained similar to those of healthy volunteers. Furthermore, we detected the MCP-1-positive cells in the interstitium of diabetic nephropathy via both immunohistochemical and in situ hybridization analyses. CONCLUSION: These observations suggest that locally produced MCP-1 may be involved in the development of advanced diabetic nephropathy, especially in the formation of tubulointerstitial lesions possibly through Mphi recruitment and activation. Moreover, up-regulation of MCP-1 may be a common pathway involved in the progressive tubulointerstitial damage in diabetic nephropathy as well as inflammatory renal diseases. PMID- 11012885 TI - Microsphere-adenoviral complexes target and transduce the glomerulus in vivo. AB - BACKGROUND: Developing new treatments for glomerulonephritis makes the glomerulus a logical target for gene therapy. Microspheres may lodge in the glomerulus, and replication-deficient recombinant adenoviruses are potent mediators of gene transfer. We postulated that adenoviral-microsphere complexes could result in DNA transfer (transduction) into glomerular cells in vivo. METHODS: Two adenoviruses, each one containing a luciferase or beta-galactosidase (beta-gal) transgene expression cassette, were complexed to polystyrene microspheres. To assess in vivo glomerular transduction with this tool, male Sprague-Dawley rats underwent aortic injections with adenovirus linked to 11 or 16 microm diameter microspheres. RESULTS: After 48 hours, adenoviral-microsphere complexes resulted in transduction of up to 19% of glomeruli per kidney section. Endothelial and mesangial cells were transduced with this approach, and transprotein expression persisted for 21 days. Transduction efficiency was greater in the 16 microm group. For all rats, there was a strong correlation between kidney luciferase levels and the number of beta-gal-positive glomeruli (r = 0.87), indicating that transgene expression was primarily glomerular in location. This was supported by reverse transcriptase in situ polymerase chain reaction, which demonstrated glomerular localization of the beta-gal transgene. CONCLUSIONS: The aortic injection of adenoviral-microsphere complexes transduces the glomerulus in vivo and may be a useful tool in developing approaches to gene therapy of glomerular disease. PMID- 11012886 TI - Hepatocyte growth factor induces branching tubulogenesis in MDCK cells by modulating the activin-follistatin system. AB - BACKGROUND: The activin-follistatin system is expressed in tubular cells of the kidney. The present study was conducted to examine the role of the activin follistatin system in tubulogenesis using Madin-Darby canine kidney (MDCK) cells as a model system. METHODS: Tubulogenesis was assessed using MDCK cells cultured in collagen gel. The effect of recombinant human activin A on tubulogenesis was examined. Blockade of the action of endogenous activin was achieved by either adding follistatin or transfection of dominant-negative mutant of the type II activin receptor. The production of activin A was examined by Northern blotting, in situ hybridization, and Western blotting. RESULTS: MDCK cells expressed mRNA for the betaA subunit of activin. These cells formed spherical cysts when cultured in collagen gel. Hepatocyte growth factor (HGF) added to the spherical cysts induced branching tubulogenesis. When activin A was added together with HGF, activin A blocked the branching tubulogenesis induced by HGF, and the activin-treated cells were scattered. Conversely, follistatin, an antagonist of activin A, induced branching tubulogenesis qualitatively similar to that induced by HGF. Adenovirus vector-mediated transfer of the gene encoding truncated type II activin receptor, which acts as a dominant negative mutant, also induced branching tubulogenesis. Finally, HGF markedly inhibited the production of activin A in MDCK cells cultured in collagen gel. CONCLUSION: Activin A produced in MDCK cells tonically inhibits branching tubulogenesis, and HGF induced branching tubulogenesis mainly by blocking the production of activin A. PMID- 11012887 TI - Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney. AB - BACKGROUND: Reabsorption of albumin from the glomerular filtrate occurs via receptor-mediated endocytosis in the proximal tubule. This process is initiated by binding of albumin in apical clathrin-coated pits, followed by endocytosis and degradation in lysosomes. Although binding sites have been characterized by kinetic studies, the receptors responsible for the binding of albumin have not been fully identified. Two giant glycoproteins, cubilin and megalin, constitute important endocytic receptors localized to the kidney proximal tubule. METHODS: In the present study, we examined the colocalization of cubilin and megalin in the endocytic pathway and the relationship between the uptake of albumin and the expression of cubilin and megalin in opossum kidney (OK) proximal tubule cells by immunocytochemistry and immunoblotting. RESULTS: OK cells expressed both cubilin and megalin. The light microscope labeling patterns for cubilin and megalin were almost identical and were mainly located at the surface area of the cells. Cubilin and megalin were also shown to colocalize on cell surface microvilli, in coated pits, and in endocytic compartments at the electron microscope level. Endocytosed bovine serum albumin (BSA) was identified exclusively in cells expressing megalin and cubilin. Uptake of BSA-FITC was saturable and inhibited by receptor-associated protein (RAP) and by intrinsic factor-vitamin B12 complex (IF B12) at high concentrations. Significant inhibition was also observed by specific antibodies to cubilin, and megalin and cubilin antisense oligonucleotides likewise significantly reduced albumin uptake. Egg albumin did not affect the uptake of BSA. CONCLUSION: The present observations suggest that the two receptors cubilin and megalin are both involved in the endocytic uptake of albumin in renal proximal tubule cells. PMID- 11012888 TI - Diabetes mellitus increases endothelin-1 gene transcription in rat kidney. AB - BACKGROUND: Mesangial cell hypertrophy and increased extracellular matrix (ECM) contribute to mesangial expansion in early progressive diabetic nephropathy. Previous studies suggest that the growth factor endothelin-1 (ET-1) is not only up-regulated in diabetes, but may mediate the effects of hyperglycemia on mesangial cell hypertrophy and ECM synthesis. In models of diabetes mellitus, the mechanisms underlying increased ET-1 peptide and mRNA remain unknown. Therefore, our purpose is to determine whether ET-1 gene activity increases in kidneys of streptozotocin (SZT)-treated rats. METHODS: Male Sprague-Dawley rats were injected with either SZT or vehicle. Parameters including glucose, body weight, 24-hour urine volume, urinary protein, and urinary ET-1 excretion were recorded. All rats were sacrificed at 12 weeks postinjection. Prepro-ET-1 mRNA from whole kidneys was determined using both RNase protection and reverse transcription polymerase chain reaction (RT-PCR). The abundance of ET-1 peptide in primary cultured mesangial cells was detected by indirect immunofluorescence following treatment with 5.6, 11.2, or 22.5 mmol/L D-glucose for 24 hours. Cellular ET-1 mRNA was measured using RT-PCR in control cells at time 0 and also following exposure to increasing concentrations of glucose for 24 hours. Rat mesangial cells were transfected with a luciferase reporter construct containing the rat ET 1 promoter (pET1. Luc), and relative ET-1 promoter activity was measured after a 24-hour exposure to 5.6 and 22.5 mmol/L of D- or L-glucose. RESULTS: After 12 weeks of hyperglycemia, diabetic rats gained less weight (344 +/- 23.9 vs. 548.75 +/- 15.08 g), had increased urinary volume (158.6 +/- 24.32 vs. 8.38 +/- 1.56 mL/day), and had marked proteinuria (101.7 +/- 12.2 vs. 14.1 +/- 2.8 mg/day) compared with controls. Total urinary ET-1 peptide increased 26.4-fold in diabetic versus control rats (17.5083 +/- 5.405 vs. 0.6635 +/- 0.343 ng/day). ET 1 mRNA extracted from whole rat kidneys was increased 2.1-fold in diabetic versus control animals. Primary cultured rat mesangial cells demonstrated a significant increase in immunofluorescence labeling of ET-1 peptide and ET-1 mRNA in response to increasing concentrations of glucose. Furthermore, transfected mesangial cells exposed to 22.5 mmol/L D-glucose showed a 1.6-fold increase in ET-1 promoter activity relative to those treated with 5.6 mmol/L glucose. CONCLUSION: Glucose increases ET-1 gene expression in the kidney of the SZT-treated rat model of diabetes mellitus. Furthermore, high glucose induces ET-1 expression in primary cultured rat mesangial cells and directly enhances ET-1 promoter activity. The greater relative increase in peptide compared with transcription suggests the potential participation of other mechanisms such as increased mRNA stability, protein stability, and/or enhanced translational efficiency. PMID- 11012889 TI - The Duffy antigen receptor for chemokines is up-regulated during acute renal transplant rejection and crescentic glomerulonephritis. AB - BACKGROUND: Recruitment of leukocytes during immune responses requires the coordinate expression of adhesion molecules in concert with chemokines and their receptors. The Duffy antigen receptor for chemokines (DARC) binds multiple chemokines and is expressed on postcapillary venules in the normal kidney. The chemokine receptor CCR5, which shares the ligand regulated upon activation, normal T-cell expressed and secreted (RANTES) with DARC, is expressed by infiltrating T cells in the renal interstitium. As DARC might be involved in the attraction of CCR5-positive cells, we studied the distribution of DARC and CCR5 in two forms of cell-mediated renal injury: renal allograft rejection and crescentic glomerulonephritis (cGN). METHODS: A total of 87 renal specimens, including 12 pretransplant biopsies, 47 transplant biopsies (Banff 1, N = 10; Banff 2, N = 19; and various other lesions N = 18), and 28 biopsies from patients with cGN, was analyzed. Immunohistochemistry for CCR5 and DARC was performed on serial sections of formalin-fixed and paraffin-embedded tissue. RESULTS: Compared with pretransplant biopsies, the mean number of DARC-positive interstitial venules was significantly increased during both transplant rejection and cGN. This was accompanied by an infiltration of CCR5-positive leukocytes. During transplant rejection, the number and distribution of CCR5-positive cells correlated with DARC-positive venules. Infiltrating CCR5-positive leukocytes were found mainly in the interstitium, often clustering around Bowman's capsules in biopsies from cGN. The number of glomerular CCR5 positive cells is low, but they are common in a subset of crescents. CONCLUSIONS: We hypothesize that the increased number of DARC-positive venules in areas of interstitial injury and the colocalization with CCR5-positive infiltrating leukocytes may indicate a role for endothelial DARC expression during leukocyte adhesion and interstitial infiltration. PMID- 11012890 TI - Small proteoglycans of normal adult human kidney: distinct expression patterns of decorin, biglycan, fibromodulin, and lumican. AB - BACKGROUND: Among the members of the small leucine-rich proteoglycan family, decorin, biglycan, and fibromodulin have been proposed to be potent modulators of transforming growth factor-beta (TGF-beta) activity, thereby playing an important role in the pathogenesis of fibrotic kidney diseases. Furthermore, decorin expression influences the expression of p21WAF1/CIP1, which has been related to kidney hypertrophy and hyperplasia. However, none of the members of this proteoglycan family have been investigated in normal adult human kidney cortex, thus making it impossible to correlate disease-mediated alterations of their expression with the normal situation in vivo. METHODS: The chondroitin/dermatan sulfate proteoglycans, decorin and biglycan, and the keratan sulfate proteoglycans, fibromodulin and lumican, were investigated in normal human adult renal cortex by immunohistochemistry on the light and electron microscopic level and by in situ hybridization. Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) methods were used to get an estimate of their expression in isolated glomeruli. Decorin excretion with the urine was measured by Western blotting. RESULTS: Two bands of decorin and a single band of biglycan mRNA were identified in Northern blots of isolated glomeruli. Amplification by RT-PCR was required to detect the signals for fibromodulin and lumican. All four proteoglycans were preferentially expressed in the renal interstitium with accumulations around tubules. Weak expression was found in the mesangial matrix. Biglycan was expressed by glomerular endothelial cells and, together with fibromodulin, was synthesized and deposited in distal tubular cells and collecting ducts. Immunogold labeling indicated the presence of the proteoglycans in the glomerular basement membrane, which was interpreted as a result of glomerular filtration. Indirect evidence suggested tubular reuptake of decorin after glomerular filtration. CONCLUSION: The data indicate that the different cells of the adult human kidney are characterized by a distinct expression pattern of the four small proteoglycans. It is suggested that these proteoglycans may have distinct pathophysiological roles depending upon whether they are expressed by mesangial cells, endothelial cells, epithelial cells, or cells of the tubulointerstitium. PMID- 11012891 TI - CD59 protects rat kidney from complement mediated injury in collaboration with crry. AB - BACKGROUND: As previously reported, the membrane-bound complement regulator at the C3 level (Crry/p65) is important in maintaining normal integrity of the kidney in rats. However, the role of a complement regulator at the C8/9 level (CD59) is not clear, especially when activation of complement occurs at the C3 level. The aim of this work was to elucidate the in vivo role of CD59 under C3 activating conditions. METHODS: Two monoclonal antibodies, 5I2 and 6D1, were used to suppress the function of Crry and CD59, respectively. In order to activate alternative the pathway of complement, the left kidney was perfused with 5I2 and/or 6D1 and was recirculated. RESULTS: In the kidneys perfused with 5I2 alone, deposition of C3 and membrane attack complex (MAC) was observed in the peritubular capillaries, vasa recta, and tubular basement membranes. Cast formation, tubular dilation and degeneration, and cellular infiltration were observed at days 1 and 4, and they recovered by day 7. Further suppression of CD59 by 6D1 significantly enhanced the deposition of MAC and worsened the already exacerbated tubulointerstitial injury. These effects of 6D1 were dose dependent. Perfusion with 6D1 alone did not induce histologic damage or MAC deposition in the tubulointerstitium. CONCLUSIONS: In rats, CD59 maintains normal integrity of the kidney in collaboration with Crry in rats against complement-mediated damage in vivo. PMID- 11012892 TI - Complement is activated in kidney by endotoxin but does not cause the ensuing acute renal failure. AB - BACKGROUND: Acute renal failure (ARF) in sepsis occurs when the release of multiple inflammatory mediators is induced by bacterial endotoxins. C3 mRNA is markedly up-regulated in mouse kidney after exposure to lipopolysaccharide (LPS). We hypothesized that LPS could induce tubular synthesis and secretion of C3, leading to activation of the complement cascade and direct renal tubular injury. METHODS: ARF was induced in mice by intravenous injection of LPS and was confirmed by an acute rise in blood urea nitrogen (BUN) and histologically by acute tubular necrosis. Three separate strategies were used to investigate the role of the complement system in this model of ARF: (1) Crry-Ig, a recombinant protein containing the potent murine complement C3 activation inhibitor Crry was injected at the same time as LPS (N = 8). (2) LPS was injected into transgenic mice overexpressing Crry in glomeruli and tubules (N = 8), and (3) LPS was injected into C3-deficient mice (N = 5). RESULTS: Compared with unmanipulated mice, C3 staining by immunofluorescence (IF) microscopy in mice injected with LPS was greater in renal cortical tubular cells (IF score of 2. 1 +/- 0.1 vs. 1.4 +/- 0.2 in controls, P = 0.013), most prominently at the basolateral surface. LPS injection led to a 16- to 42-fold increase in urinary C3 excretion. Despite reduction or complete elimination of renal C3 with maneuvers suppressing complement activation, BUN values were not statistically different across all groups. In no experiment did BUN values correlate with the extent of C3 staining. CONCLUSION: Although LPS up-regulates renal C3 synthesis, resulting in basolateral tubular C3 deposition, this is not responsible for LPS-induced ARF in mice. PMID- 11012893 TI - Elastic fiber proteins in the glomerular mesangium in vivo and in cell culture. AB - BACKGROUND: Glomerular capillaries of the mammalian kidney are exposed to high intraluminal hydrostatic pressures and require elastic constraint to maintain size, shape, and integrity. Previous morphological and functional studies indicated that the extracellular matrices of glomeruli, that is, basement membrane and mesangial matrix, contribute to glomerular resilience and mechanical stability. Immunofluorescence microscopy findings demonstrated elastic fiber components to be located in the renal vasculature, including glomeruli. The aim of this study was to clarify the exact glomerular localization, composition, and cellular production of these proteins. METHODS: We examined the renal distribution of the elastic fiber proteins fibrillin-1, emilin, microfibril associated glycoproteins (MAGPs) 1 and 2, latent transforming growth factor binding protein-1 (LTBP-1), and elastin using immunohistology and immunoelectron microscopy of human, rat, and mouse kidneys. In mesangial cell cultures, we also studied the expression and extracellular deposition of such proteins by use of Northern blotting and immunocytochemistry. RESULTS: Fibrillin-1, emilin, MAGPs 1 and 2, and LTBP-1 were present in glomeruli of mouse, rat, and human kidney, where they were located predominantly in the mesangial extracellular matrix underlying glomerular endothelium and basement membrane. Several of these proteins, as well as elastin, were also expressed in the renal vasculature. While elastin localized to the glomerular vascular pole in afferent and efferent arterioles extending to Bowman's capsule, it was not found in the glomerular capillary tuft. Cultured mesangial cells of rat, mouse, and human kidneys expressed mRNAs of fibrillin-1, emilin, MAGP-2, and elastin, and the respective proteins localized within and outside of mesangial cells, as shown by immunocytochemistry. mRNA expression of fibrillin-1, emilin, and elastin was strong in quiescent mesangial cells; their gene expression was further up regulated by transforming growth factor-beta1, while it was transiently reduced when cells were exposed to mitogenic 10% fetal calf serum and platelet-derived growth factor. CONCLUSIONS: These findings demonstrate that specific elastic fiber proteins are produced and secreted by mesangial cells. This process is regulated by growth factors. Their abundance in the extracellular matrix of the mesangium is in keeping with the concept that elastic fiber proteins contribute to the mechanical stability and elastic strength of the glomerular capillary tuft. PMID- 11012894 TI - Expression of DNA topoisomerases in chronic proliferative kidney disease. AB - BACKGROUND: Circulating autoantibodies to human topoisomerases have been reported in glomerular kidney disease associated with scleroderma and systemic lupus erythematosus. However, limited information is available about the expression of topoisomerases in the kidney under normal and pathological conditions. METHODS: The expression of DNA topoisomerases I and IIalpha was studied by immunohistochemistry on archival biopsies from 70 patients with chronic renal diseases. Normal kidney tissue was examined for comparison. Topoisomerase I was detected by means of monoclonal antibody (mAb) C21, and topoisomerase IIalpha was detected by means of mAb Ki-S4. In addition, mAb Ki-M1p was used to assess the density of monocytic infiltrates. All parameters were assessed in a semiquantitative manner. RESULTS: Glomerular topoisomerase IIalpha levels were increased in mesangial proliferative glomerulonephritis (MPGN), rapidly progressive glomerulonephritis (RPGN), and lupus nephritis (LN) and were reduced in membranous glomerulonephritis (MGN), chronic transplant nephropathy (CTN), and tubulointerstitial nephritis (TIN). Tubular epithelia displayed high topoisomerase IIalpha levels in mesangiocapillary glomerulonephritis (MCGN), RPGN, TIN, miscellaneous entities (MISC) and LN, and displayed low levels in MPGN and CTN. Topoisomerase I expression was high in the glomeruli of focal segmental glomerulosclerosis (FSGS), MCGN, and RPGN and was extreme in LN, whereas it was strikingly diminished in the glomeruli of MGN, CTN, and TIN. Almost all conditions displayed lower tubular topoisomerase I levels than normal kidney, except for LN, in which the enzyme content was markedly increased. Increased glomerular monocytic infiltrates were found in FSGS, MCGN, RPGN, TIN, and LN, and tubulointerstitial Ki-M1p+ cells were seen at high numbers in MCGN, RPGN, TIN, MISC, and LN. The expression of the topoisomerases I and IIalpha was significantly correlated; also, topoisomerases showed a positive association with the density of monocytic infiltrates. The parameter profiles exhibited significant differences between distinct types of chronic renal disease. CONCLUSION: Topoisomerase IIalpha expression is tightly linked to cell cycling, and topoisomerase I is likely a reflection of gene transcription. Rapidly progressing glomerular disease therefore appears to be accompanied by active mesangial cell proliferation and increased metabolic activity in glomerular cells. The correlation with inflammatory infiltrates is likely to reflect a positive feedback mechanism involving cytokines, growth factors, and adhesion molecules. Assessment of topoisomerases may therefore be of diagnostic help and might allow prognostic predictions. Provided that our observations are supported by clinicopathological follow-up studies, one might envisage the use of topoisomerase inhibitors in the therapy of chronic proliferative renal disease refractory to current treatment protocols. PMID- 11012895 TI - Binding properties of a selective tritiated vasopressin V2 receptor antagonist, [H]-SR 121463. AB - BACKGROUND: [3H]-SR 121463 is the first radiolabeled selective nonpeptide vasopressin V2 receptor antagonist ligand that has been reported to date. In the present work, we studied the binding properties of [3H]-SR 121463 for renal V2 receptors from animal and human origins. METHODS: Binding studies were performed with [3H]-SR 121463 in Chinese hamster ovary (CHO) cells transfected with the human V2 receptor and in various kidney preparations expressing the native V2 receptors (rat, rabbit, dog, pig, monkey, and human). Autoradiographies were performed in rat and human kidney sections. RESULTS: [3H]-SR 121463 binding to CHO cells stably transfected with the cloned human renal V2 receptor was specific, highly stable, time dependent, saturable, and reversible. A single population of high-affinity binding sites was identified (Kd = 0.94 +/- 0.34 nmol/L, Bmax = 9876 +/- 317 fmol/mg protein). Of note, [3H]-SR 121463 revealed a higher number (about 40%) of V2 sites than [3H]-AVP in the same preparation. Displacement of [3H]-SR 121463 binding by reference peptide and nonpeptide vasopressin/oxytocin compounds exhibited a typical AVP V2 profile. [3H]-SR 121463 also displayed a high affinity for native V2 receptors in several kidney preparations from rat, pig, dog, rabbit, bovine, monkey, and human. The autoradiographic experiments using rat and human kidney sections showed intense labeling in the medullopapillary region and lower intensity in the cortex, consistent with a main localization of V2 receptors on collecting tubules. CONCLUSION: [3H]-SR 121463 is a useful ligand for the specific labeling of animal and human V2 receptors and could be a suitable probe for the search and in situ localization of V2 sites. PMID- 11012896 TI - Chronic effect of parathyroid hormone on NHE3 expression in rat renal proximal tubules. AB - BACKGROUND: The most abundant Na+/H+ exchanger in the apical membrane of proximal tubules is the type 3 isoform (NHE3), and its activity is acutely inhibited by parathyroid hormone (PTH). In the present study, we investigate whether changes in protein abundance as well as in mRNA levels play a significant role in the long-term modulation of NHE3 by PTH. METHODS: Three groups of animals were compared: (1) HP: animals submitted to hyperparathyroidism by subcutaneous implantation of PTH pellets, providing threefold basal levels of this hormone (2.1 U. h-1); (2) control: sham-operated rats in which placebo pellets were implanted; (3) PTX: animals submitted to hypoparathyroidism by thyroparathyroidectomy followed by subcutaneous implantation of thyroxin pellets, which provided basal levels of thyroid hormone. After eight days, we measured bicarbonate reabsorption in renal proximal tubules by in vivo microperfusion. NHE3 activity was also measured in brush border membrane (BBM) vesicles by proton dependent uptake of 22Na. NHE3 expression was evaluated by Northern blot, Western blot and immunohistochemistry. RESULTS: Bicarbonate reabsorption in renal proximal tubules was significantly decreased in HP rats. Na+/H+ exchange activity in isolated BBM vesicles was 6400 +/- 840, 9225 +/- 505, and 12205 +/- 690 cpm. mg-1. 15 s-1 in HP, sham, and PTX groups, respectively. BBM NHE3 protein abundance decreased 39.3 +/- 8.2% in HP rats and increased 54.6 +/- 7.8% in PTX rats. Immunohistochemistry showed that expression of NHE3 protein in apical BBM was decreased in HP rats and was increased in PTX rats. Northern blot analysis of total kidney RNA showed that the abundance of NHE3 mRNA was 20.3 +/- 1.3% decreased in HP rats and 27. 7 +/- 2.1% increased in PTX. CONCLUSIONS: Our results indicate that the chronic inhibitory effect of PTH on the renal proximal tubule NHE3 is associated with changes in the expression of NHE3 mRNA levels and protein abundance. PMID- 11012897 TI - Microcomputed tomography of kidneys following chronic bile duct ligation. AB - BACKGROUND: In hepatic cirrhosis, renal sodium and water retention can occur prior to decreases in renal blood flow (RBF). This may be explained in part by redistribution of the intrarenal microcirculation toward the juxtamedullary nephrons. To appreciate this three-dimensional spatial redistribution better, we examined the intrarenal microcirculatory changes using microcomputed tomography (micro-CT) in rats subjected to chronic bile duct ligation (CBDL). METHODS: Six kidneys from control rats and eight kidneys from rats that had undergone CBDL for 21 days were perfusion fixed in situ at physiological pressure, perfused with silicon-based Microfil containing lead chromate, embedded in plastic, and scanned by micro-CT. The microvasculature in the reconstructed three-dimensional renal images was studied using computerized image-analysis techniques. To determine the physiological condition of the rats, parallel experiments were conducted on six control and six CBDL rats to measure mean arterial pressure (MAP), RBF, glomerular filtration rate (GFR), urine flow (UF) rate, and sodium excretion by conventional methods. RESULTS: The percentage of vasculature in the renal cortex from CBDL rats was significantly decreased (10.8 +/- 0.4% vs. 16.8 +/- 2.7% control values). However, the vascular volume fractions of the medullary tissues were not significantly altered. There were no significant differences in the number of glomeruli between groups (36,430 +/- 1908 CBDLs, 36,609 +/- 3167 controls). The CBDL rats had a similar GFR than the controls but a reduced MAP, RBF, UF, and sodium excretion. CONCLUSIONS: The results indicate that after CBDL, there is a selective decrease in cortical vascular filling, which may contribute to the salt and water retention that accompanies cirrhosis. PMID- 11012898 TI - H+-ATPase activity on unilateral ureteral obstruction: interaction of endogenous nitric oxide and angiotensin II. AB - BACKGROUND: A number of cytokines, vasoactive compounds, chemoattractant molecules, and growth factors are up-regulated in obstruction. Following the onset of ureteral obstruction, angiotensin II production is rapidly stimulated. Cytokine-induced expression of inducible nitric oxide synthase (iNOS) has been reported in primary cultures of inner medullary collecting duct (IMCD) cells. We found that the defective urinary acidification in unilateral ureteral obstruction (UUO) includes an intensive decrease in bafilomycin-sensitive H+-ATPase activity in microdissected IMCD segments. METHODS: To investigate the interaction between endogenous nitric oxide and angiotensin II on H+-ATPase activity, we used microdissected IMCD segments of unilaterally obstructed, contralateral, and control kidneys to measure the bafilomycin-sensitive ATPase activity and nitric oxide synthase (NOS) activity. The generated NO was also evaluated. RESULTS: Preincubation of obstructed IMCD segments in the presence of a competitive inhibitor of NOS, NG-nitro-L-arginine methyl ester (L-NAME) 1 mmol/L, and in the presence of a specific inhibitor of calcium/calmodulin-independent NOS (iNOS), aminoguanidine 1 mmol/L, each for 60 minutes, significantly increased bafilomycin sensitive H+-ATPase. A greater increase on iNOS activity (fmol [3H] citrulline/min/microg protein) and a lesser increase in calcium/calmodulin dependent NOS activity (cNOS) were observed in the obstructed renal medulla. This inhibitory effect of obstruction was abolished when IMCDs were incubated with 10 5 to 10-8 mol/L losartan. Decreasing doses of the angiotensin II type 1 (AT1) receptor inhibitor caused an increase in bafilomycin-sensitive H+-ATPase, with a maximum increase at 10-8 mol/L losartan. A decrease on iNOS activity was demonstrated in the obstructed renal medulla incubated with losartan in concentrations of 10-5 to 10-8 mol/L, the same losartan concentrations that showed recovery of vacuolar H+-ATPase activity. Similarly, a decrease on the generation of NO after incubation with losartan 10-5 to 10-8 mol/L was shown. CONCLUSION: From these results, we suggest that endogenous NO increased by iNOS is involved in the inhibition of H+-ATPase activity in obstructed IMCD segments. The recovery of H+-ATPase activity in IMCD of obstructed kidneys induced by losartan may be related to a decrease of inducible NOS activity. PMID- 11012899 TI - Cyclosporine stimulates Na+-K+-Cl- cotransport activity in cultured mouse medullary thick ascending limb cells. AB - BACKGROUND: Cyclosporine (CsA) has been shown to alter the activity of plasma membrane transporters in kidney epithelial cells. In this study, we have investigated the effects of CsA on Na+,K+-ATPase and Na+-K+-Cl- cotransport activities in cultured cells derived from microdissected mouse medullary thick ascending limb (mTAL) cells. METHODS: Experiments were carried out on subcultured confluent mouse TAL cells. Reverse transcription-polymerase chain reaction experiments showed that they expressed the mNKCC2 electroneutral Na+-K+-Cl- cotransporter and ROM-K1 and ROMK2 potassium channel mRNA. Western blotting also revealed the presence of the 40 kD ROMK protein using an anti-ROMK antibody. The effect of CsA (100 ng/mL) on ion transport was assessed by measuring the influx and efflux of rubidium (86Rb+) and 36Cl-, used as tracers of K+ and Cl- movements, on cells grown on Petri dishes or permeable filters. RESULTS: CsA inhibited by 38% the ouabain-sensitive component of 86Rb+ influx mediated by the Na+,K+-ATPase pumps. CsA also increased by 38% the ouabain-resistant furosemide sensitive component (Or-Fs) of 86Rb+ influx, reflecting the Na+-K+-Cl- cotransport activity and stimulated the basolateral efflux of 36Cl- from mTAL cells grown on filters. The CsA-stimulated basal efflux of Cl- was prevented by the basal addition of the Cl- channel blocker 5-nitro-2-(3-phenylpropylamino) benzoate (NPPB, 10-4 mol/L). Apical addition of the K+ channel blocking agent Ba2+ (10-4 mol/L) partially prevented the CsA-stimulated basal efflux of Cl-. Adding Ba2+ to the luminal side of cells grown on Petri dishes also prevented the rise in apical 86Rb+ efflux and the increased Or-Fs component of 86Rb+ influx caused by CsA. CONCLUSION: These results indicated that CsA may stimulate the Na+ K+-Cl- cotransport activity and also suggested that this immunosuppressive agent may interfere in the recycling of apical K+ in this model of cultured mouse TAL cells. PMID- 11012900 TI - Proinflammatory gene expression and macrophage recruitment in the rat remnant kidney. AB - BACKGROUND: Macrophage (Mphi) infiltration may contribute to chronic renal injury. We therefore sought to examine the expression of genes associated with Mphi recruitment in the rat remnant kidney model. METHODS: Male Munich Wistar rats underwent 5/6 nephrectomy or sham operation (SHM, N = 18) and received no treatment (VEH, N = 18), enalapril 100 mg/L (ENA, N = 18), or candesartan 70 mg/L (CSN, N = 24) in drinking water. Competitive, quantitative reverse transcription polymerase chain reaction was used to determine renal cortex mRNA levels for cell adhesion molecules vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), the Mphi chemoattractant monocyte chemoattractant protein-1 (MCP-1), Mphi products interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), and the profibrotic cytokine transforming growth factor beta1 (TGF-beta1), at intervals post-nephrectomy. RESULTS: Glomerular and interstitial Mphi infiltration in VEH rats was associated with an early (4 week) and sustained rise in MCP-1 and TGF-beta1 mRNA levels. Progressive increases in ICAM-1, VCAM-1, IL-1beta, and TNF-alpha expression followed at 8 and 12 weeks. Immunostaining in VEH rats localized TGF-beta1 to glomeruli, tubules, and interstitium; MCP-1 to tubules and interstitial cells; ICAM-1 to glomeruli; and IL-1beta and TNF-alpha to tubules and interstitial cells. At 12 weeks, both treatments normalized systolic blood pressure (ENA, 105 +/- 6; CSN, 97 +/- 3 mm Hg) and the urinary protein excretion rate (ENA, 8.4 +/- 0.9; CSN, 5.7 +/- 0.8 mg/day), prevented renal injury (focal and segmental glomerulosclerosis: ENA, 3.3 +/- 0.9; CSN, 1.3 +/- 0.4%), and suppressed Mphi infiltration and cytokine expression (with the exception of TNF-alpha) to near SHM levels. CONCLUSIONS: These findings support the hypothesis that the coordinated up-regulation of several molecules regulating Mphi recruitment and activation is a fundamental response to renal mass ablation and is dependent on an intact renin-angiotensin system. We speculate that these responses may play a role in the pathogenesis of the ensuing glomerulosclerosis and tubulointerstitial fibrosis. PMID- 11012901 TI - Effects of diabetes and hypertension on glomerular transforming growth factor beta receptor expression. AB - BACKGROUND: Several studies have suggested that transforming growth factor-beta1 (TGF-beta1) is an important determinant of diabetic glomerular injury. TGF-beta1 forms a heteromeric complex with two cellular receptor subtypes, designated TGF beta RII and TGF-beta RI, but the effects of diabetes mellitus on glomerular TGF beta receptor expression have not been completely elucidated. We first compared the effect of experimental type I diabetes mellitus and uninephrectomy on glomerular TGF-beta receptor expression in spontaneously hypertensive rats (SHRs), and then sought to determine whether changes in TGF-beta receptor expression were strain specific by studying normotensive Wistar-Kyoto (WKY) rats. METHODS: Five groups of male SHRs were studied. The first group received streptozotocin (60 mg/kg IV) and was studied after one week. The second group received streptozotocin and was studied after two weeks. The third group received streptozotocin (60 mg/kg IV) but received insulin to maintain euglycemia. The fourth group of age-matched SHRs served as the control group, while a fifth group of SHRs underwent uninephrectomy. Four groups of male WKY rats were also studied. The first group of WKY rats served as the age-matched control group. The second group of WKY rats received streptozotocin, while a third group of WKY rats underwent uninephrectomy. The fourth group underwent uninephrectomy and received streptozotocin. At each time point, glomeruli were isolated for protein extraction, and the protein was subjected to Western blot analysis of TGF-beta RII and TGF-beta RI expression. RESULTS: Basal expression of both TGF-beta receptors per microgram of glomerular protein was similar in normotensive WKY rats and hypertensive SHRs. Hyperglycemia (blood glucose level, 17.8 +/- 2.9 mmol/L) led to an early twofold increase in TGF-beta RII protein expression and a fourfold increase in TGF-beta RI protein expression in the glomeruli of hypertensive diabetic SHRs compared with euglycemic SHRs (blood glucose level, 5.8 +/- 0.8 mmol/L), which was sustained after two weeks. Insulin treatment (blood glucose level, 5. 2 +/- 0.9 mmol/L) normalized both TGF-beta RII and TGF beta RI expression in the glomeruli of SHRs that received streptozotocin. Glomerular capillary hypertension in the uninephrectomized SHRs led to a twofold increase in glomerular TGF-beta RII protein expression, but did not reproduce the effect of diabetes mellitus on TGF-beta RI expression. In contrast to the findings in SHRs, neither hyperglycemia (blood glucose level, 15.5 +/- 2.1 mmol/L), uninephrectomy, nor hyperglycemia (blood glucose level, 16.8 +/- 3.0 mmol/L) and uninephrectomy altered TGF-beta receptor expression in the glomeruli of normotensive WKY rats. CONCLUSION: These studies support the hypothesis that hemodynamic factors and metabolic factors influence glomerular TGF-beta receptor in vivo in the SHRs. PMID- 11012902 TI - Ryanodine receptor and capacitative Ca2+ entry in fresh preglomerular vascular smooth muscle cells. AB - BACKGROUND: A multiplicity of hormonal, neural, and paracrine factors regulates preglomerular arterial tone by stimulating calcium entry or mobilization. We have previously provided evidence for capacitative (store-operated) Ca2+ entry in fresh renal vascular smooth muscle cells (VSMCs). Ryanodine-sensitive receptors (RyRs) have recently been identified in a variety of nonrenal vascular beds. METHODS: We isolated fresh rat preglomerular VSMCs with a magnetized microsphere/sieving technique; cytosolic Ca2+ ([Ca2+]i) was measured with fura-2 ratiometric fluorescence. RESULTS: Ryanodine (3 micromol/L) increased [Ca2+]i from 79 to 138 nmol/L (P = 0.01). Nifedipine (Nif), given before or after ryanodine, was without effect. The addition of calcium (1 mmol/L) to VSMCs in calcium-free buffer did not alter resting [Ca2+]i. In Ca-free buffer containing Nif, [Ca2+]i rose from 61 to 88 nmol/L after the addition of the Ca2+-ATPase inhibitor cyclopiazonic acid and to 159 nmol/L after the addition of Ca2+ (1 mmol/L). Mn2+ quenched the Ca/fura signal, confirming divalent cation entry. In Ca-free buffer with Nif, [Ca2+]i increased from 80 to 94 nmol/L with the addition of ryanodine and further to 166 nmol/L after the addition of Ca2+ (1 mmol/L). Mn2+ quenching was again shown. Thus, emptying of the sarcoplasmic reticulum (SR) with ryanodine stimulated capacitative Ca2+ entry. CONCLUSION: Preglomerular VSMCs have functional RyR, and a capacitative (store-operated) entry mechanism is activated by the depletion of SR Ca2+ with ryanodine, as is the case with inhibitors of SR Ca2+-ATPase. PMID- 11012903 TI - Changes in conjugated linoleic acid and its metabolites in patients with chronic renal failure. AB - BACKGROUND: Conjugated linoleic acid (CLA) is a mixture of isomers of linoleic acid with conjugated double bonds that constitutes the most abundant fatty acid with conjugated dienes (CDs) in humans. CLA, erroneously considered in the past as a product of lipoperoxidation, has a dietary origin and has shown to possess anticarcinogenic and anti-atherogenic activity, mainly in animal studies. CLA can be metabolized to conjugated linolenic acid (CD18:3) and to conjugated eicosatrienoic acid (CD20:3) and these metabolites may be implicated in CLA activity. Because of the presence of dyslipidemia and the high incidence of cardiovascular and neoplastic diseases in uremic patients, we evaluated CLA and its metabolites in these patients in order to evaluate their metabolism and site distribution. METHODS: We measured CLA, CD18:3, CD20:3, CD fatty acid hydroperoxides (lipoperoxidation products), and linoleic acid in the plasma, adipose tissue, and red blood cell (RBC) membranes by using high-pressure liquid chromatography in the following groups: (1) 23 chronic renal failure (CRF) patients with creatine clearance (CCr)> 10 mL/min (26.2 +/- 16.7); (2) 21 end stage CRF patients in conservative treatment with CCr <10 mL/min (6.8 +/- 1.8); (3) 30 hemodialysis (HD) patients; and (4) 30 healthy controls. RESULTS: The incorporation of CLA, CD18:3, and CD20:3 in RBC membranes was significantly reduced in group 1 and was even more reduced in groups 2 and 3. CLA significantly increased both in the plasma and adipose tissue of end-stage CRF patients only. CD18:3 and CD20:3 did not change in the plasma and adipose tissue of any group. No significant changes in linoleic acid and CD fatty acid hydroperoxides were found. CONCLUSIONS: The alterations of CD in CRF patients are not due to lipoperoxidation. The increased levels of CLA in plasma and adipose tissue of end stage CRF patients may be due either to a reduced metabolization of CLA to CD18:3 and CD20:3, or to an altered site distribution with reduced incorporation in cellular membranes and accumulation in the plasma and adipose tissue. The clinical significance of these changes remains to be investigated. PMID- 11012904 TI - Inflammation and microalbuminuria in nondiabetic and type 2 diabetic subjects: The Insulin Resistance Atherosclerosis Study. AB - BACKGROUND: Microalbuminuria is a risk factor for cardiovascular disease, but the underlying pathomechanisms are still poorly understood. A relationship between C reactive protein (CRP), a sensitive marker of inflammation, and atherosclerotic disease has been reported recently. METHODS: We hypothesized that microalbuminuria might be associated with chronic inflammation and investigated the relationship of urinary albumin excretion, as assessed from the albumin-to creatinine ratio (ACR), in an untimed morning urine specimen, and two inflammatory markers (CRP and fibrinogen) in the large, triethnic population of the Insulin Resistance Atherosclerosis Study (IRAS). After exclusion of subjects with macroalbuminuria, 1481 subjects were studied. RESULTS: Both inflammatory markers were related to urinary ACR (r = 0.17 for CRP and r = 0.14 for fibrinogen, both P = 0.0001), an association that remained significant after adjustment for demographic variables, diabetic status, smoking, and use of angiotensin-converting enzyme inhibitors (P < 0.01). Mean levels of CRP and fibrinogen were elevated in microalbuminuric (N = 262) versus normoalbuminuric (N = 1219) subjects (5.37 +/- 0.47 vs. 3.80 +/- 0.15 mg/L and 295.7 +/- 4. 0 vs. 278.2 +/- 1.6 mg/dL, both P < 0.0001). The associations were consistent among nondiabetic and type 2 diabetic subjects and among the three ethnic groups of the IRAS (non-Hispanic whites, blacks, Hispanics). In a logistic regression model, fibrinogen was independently associated with microalbuminuria (P = 0.047), along with hypertension, female gender, waist circumference, and fasting blood glucose, while CRP was not independently related to microalbuminuria in this model (P = 0.26). CONCLUSION: We have shown an association of CRP and fibrinogen with urinary albumin excretion in the microalbuminuric range in type 2 diabetic and nondiabetic individuals. Chronic inflammation therefore emerges as a potential mediator between microalbuminuria and macrovascular disease. PMID- 11012905 TI - Renal function and blood pressure five years after puumala virus-induced nephropathy. AB - BACKGROUND: Nephropathia epidemica (NE) is a mild form of hemorrhagic fever with renal syndrome caused by Puumala hantavirus. Its long-term prognosis is considered favorable. Some reports suggest, however, that a previous hantavirus infection increases the risk of hypertension. METHODS: We studied 46 previously healthy subjects (26 males and 20 females, mean age of 44 years) who had serologically confirmed NE three to seven years previously, and 38 healthy, seronegative controls (22 males and 16 females, mean age of 44 years). Ambulatory blood pressure (ABP) was monitored. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were determined by 51CrEDTA and 131I-hippurate clearances, respectively. The filtration fraction (FF) was calculated. Quantitative 24-hour urinary protein excretion (UprotE) and timed overnight urinary excretion of alpha1-microglobulin were measured. RESULTS: The NE patients had a higher mean ambulatory systolic BP than the controls (123 +/- 13 vs. 117 +/ 9 mm Hg, P = 0. 008). GFR and FF were increased in patients compared with controls (GFR, 120 +/- 20 vs. 109 +/- 14 mL/min/1.73 m2, P = 0.006; FF, 19 +/- 3 vs. 18 +/- 3%, P = 0.030), but ERPF did not differ between the groups. The patients also had higher UPE than the controls (median 0. 18 g/day, range 0.12 to 0.38 vs. median 0.14 g/day, range 0.09 to 0. 24, P < 0.001, respectively). The overnight urinary excretion rate of alpha1-microglobulin exceeded 7 microg/min in nine patients. CONCLUSION: Three to seven years after NE, the patients had higher GFR and FF, more proteinuria, and higher ambulatory systolic BP compared with the healthy controls. NE may thus cause mild renal lesions and alterations in BP in some patients. PMID- 11012906 TI - Causes of albuminuria in patients with type 2 diabetes without diabetic retinopathy. AB - BACKGROUND: The causes of albuminuria in patients with type 2 diabetes are heterogeneous and are scantily investigated, particularly if the patient has a lack of diabetic retinopathy. Therefore, we evaluated the structural background of albuminuria in a large consecutive group of Caucasian patients with type 2 diabetes without retinopathy. METHODS: Three hundred forty-seven consecutive patients with type 2 diabetes with persistent albuminuria (>300 mg/24 h) were recorded. Fundus photo (80%) and ophthalmoscopy were performed. Ninety-three (27%) had no retinopathy, and a kidney biopsy was performed in 52 (56%) of these patients. An insufficient tissue sample was obtained in one patient. The biopsies were evaluated by three masked nephropathologists. RESULTS: The biopsies revealed diabetic glomerulopathy in 69% of the patients (28 males and 7 females), while the remaining 31% (95% CI, 18 to 44) had either nondiabetic glomerulopathies such as glomerulonephritis (N = 7, 6 males and 1 female, 13%) or normal glomerular structure (N = 9, 7 males and 2 females, 18%). No significant differences in sex, age (56 +/- 8 vs. 53 +/- 10 years, mean SD), body mass index (30 +/- 4 vs. 31 +/- 8 kg/m2), known duration of diabetes (6 +/- 6 vs. 4 +/- 3 years), GFR (95 +/- 29 vs. 89 +/- 31 mL/min/1.73 m2), albuminuria (1304 +/- 169 to 4731 vs. 1050 +/- 181 to 5176 mg/24 hours), blood pressure (150/87 +/- 16/9 vs. 145/89 +/- 16/9 mm Hg), prevalence of hypertension (89 vs. 100%), hemoglobin A1c (8.2 +/- 1.6% vs. 9.0 +/ 2.5%), and serum total cholesterol (7.1 +/- 2.4 vs. 6.3 +/- 1.6 mmol/L) were found between patients with and without diabetic glomerulopathy. CONCLUSIONS: Albuminuric patients with type 2 diabetes without diabetic retinopathy have a prevalence of biopsies with normal glomerular structure or nondiabetic kidney diseases of approximately 30%. A separation between diabetic and nondiabetic glomerular lesions was not possible based on demographic, clinical, or laboratory data. Consequently, such patients may require further evaluation, including a kidney biopsy. PMID- 11012907 TI - A modern approach to selectivity of proteinuria and tubulointerstitial damage in nephrotic syndrome. AB - BACKGROUND: The selectivity of proteinuria, introduced in clinical nephrology in 1960 and useful in predicting steroid responsiveness in nephrotic syndrome, found little place in clinical practice in subsequent decades, since its assessment did not appear to help predict histologic diagnosis or determine prognosis. The amount of proteinuria and the degree of tubulointerstitial damage appeared to be better predictors of functional outcome. A correlation between them has been found, referred to some toxicity of proteinuria on tubular cells, but so far no single feature or component of proteinuria has been identified as being responsible for this toxicity. METHODS: We evaluated 89 patients with nephrotic syndrome [9 with minimal change disease (MCD), 29 with primary focal segmental glomerulosclerosis (FSGS), and 51 with idiopathic membranous glomerulonephritis (MGN)] to determine if the selectivity of proteinuria was associated with tubulointerstitial damage. A semiquantitative grading of histologic lesions and qualitative evaluation of the "tubular" component of proteinuria expressed as a pattern of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and as fractional excretion of the low molecular weight (LMW) protein alpha1 microglobulin (FE alpha1m) were used. A second aim of the study was to assess the predictive value on functional outcome [remission or progression to chronic renal failure (CRF)] and response to therapy of the selectivity of proteinuria, considered alone and in combination with FE alpha1m. RESULTS: Proteinuria was classified as highly selective [selectivity index (SI) < or = 0.10, N = 15], moderately selective (SI > or = 0.11 < or = 0.20, N = 34), or nonselective (SI > or = 0.21, N = 40). A significant relationship was found between the SI and the histologic degree of tubulointerstitial damage (score 0 to 1 vs. score > or =2, P = 0.000), severity of the tubular component of proteinuria (mixed SDS-PAGE pattern with LMW proteins not lower than 23 kD vs. mixed pattern with LMW proteins up to 20 to 10 kD, P = 0.000), and FE alpha1m (values below vs. above a defined cut-off, P = 0.000). The functional outcome was evaluated in 60 patients with baseline normal renal function (serum creatinine 0.97 +/- 0.19 mg/dL). The patients with high, moderate, or nonselective proteinuria had 100, 50, and 29% of complete or partial remission (P = 0.0001) and 0, 25, and 35% of progression to CRF, respectively (P = 0.050). In 45 patients with moderately selective (N = 28) and nonselective (N = 17) proteinuria, according to some arbitrary cutoffs for FE alpha1m (MGN, < or = vs. > 0. 240% of creatinine clearance; FSGS and MCD, < or = vs. > 0.350%), the remission rate was 62 versus 6% in patients with FE alpha1m below or above the cutoffs (P = 0.0001), and progression to CRF was 7 and 69%, respectively (P = 0.0001). The response to therapy (complete or partial remission at the last observation), evaluated retrospectively in 40 patients, was 100, 67, and 33% in high, moderate, and nonselective proteinuria (P = 0.0002); in 30 patients with moderate and nonselective proteinuria, according to an FE alpha1m value that was < or = or > the cutoffs, the response rate was 75 versus 10% (P = 0.001). CONCLUSIONS: There is a significant relationship between selectivity of proteinuria and tubulointerstitial damage. Moreover, the selectivity of proteinuria has a predictive value on functional outcome. When proteinuria is highly selective, the tubulointerstitial damage is rather infrequent, and 100% of patients develop clinical remission. When proteinuria is moderately selective or nonselective, increasing numbers of patients develop tubulointerstitial damage; in these patients, the functional outcome and response to therapy is partly dependent on tubulointerstitial involvement, and the best predictor of functional outcome is the combination of SI and FE alpha1m. PMID- 11012908 TI - Serum C-peptide concentrations poorly phenotype type 2 diabetic end-stage renal disease patients. AB - BACKGROUND: A homogeneous patient population is necessary to identify genetic factors that regulate complex disease pathogenesis. In this study, we evaluated clinical and biochemical phenotyping criteria for type 2 diabetes in end-stage renal disease (ESRD) probands of families in which nephropathy is clustered. C peptide concentrations accurately discriminate type 1 from type 2 diabetic patients with normal renal function, but have not been extensively evaluated in ESRD patients. We hypothesized that C-peptide concentrations may not accurately reflect insulin synthesis in ESRD subjects, since the kidney is the major site of C-peptide catabolism and would poorly correlate with accepted clinical criteria used to classify diabetics as types 1 and 2. METHODS: Consenting diabetic ESRD patients (N = 341) from northeastern Ohio were enrolled. Clinical history was obtained by questionnaire, and predialysis blood samples were collected for C peptide levels from subjects with at least one living diabetic sibling (N = 127, 48% males, 59% African Americans). RESULTS: Using clinical criteria, 79% of the study population were categorized as type 1 (10%) or type 2 diabetics (69%), while 21% of diabetic ESRD patients could not be classified. In contrast, 98% of the patients were classified as type 2 diabetics when stratified by C-peptide concentrations using criteria derived from the Diabetes Control and Complications Trial Research Group (DCCT) and UREMIDIAB studies. Categorization was concordant in only 70% of ESRD probands when C-peptide concentration and clinical classification algorithms were compared. Using clinical phenotyping criteria as the standard for comparison, C-peptide concentrations classified diabetic ESRD patients with 100% sensitivity, but only 5% specificity. The mean C-peptide concentrations were similar in diabetic ESRD patients (3.2 +/- 1.9 nmol/L) and nondiabetic ESRD subjects (3.5 +/- 1.7 nmol/L, N = 30, P = NS), but were 2.5-fold higher compared with diabetic siblings (1.3 +/- 0.7 nmol/L, N = 30, P < 0.05) with normal renal function and were indistinguishable between type 1 and type 2 diabetics. Although 10% of the diabetic ESRD study population was classified as type 1 diabetics using clinical criteria, only 1.5% of these patients had C peptide levels less than 0.20 nmol/L, the standard cut-off used to discriminate type 1 from type 2 diabetes in patients with normal renal function. However, the criteria of C-peptide concentrations> 0.50 nmol/L and diabetes onset in patients who are more than 38 years old identify type 2 diabetes with a 97% positive predictive value in our ESRD population. CONCLUSIONS: Accepted clinical criteria, used to discriminate type 1 and type 2 diabetes, failed to classify a significant proportion of diabetic ESRD patients. In contrast to previous reports, C-peptide levels were elevated in the majority of type 1 ESRD diabetic patients and did not improve the power of clinical parameters to separate them from type 2 diabetic or nondiabetic ESRD subjects. Accurate classification of diabetic ESRD patients for genetic epidemiological studies requires both clinical and biochemical criteria, which may differ from norms used in diabetic populations with normal renal function. PMID- 11012909 TI - Effects of bicarbonate- and lactate-buffered replacement fluids on cardiovascular outcome in CVVH patients. AB - BACKGROUND: Bicarbonate-buffered replacement fluid (RF-bic) in continuous venovenous hemofiltration (CVVH) may be superior to lactate-buffered replacement fluid (RF-lac) in acute renal failure. In an open, randomized, multicenter study, we investigated the effects of RF-bic and RF-lac on cardiovascular outcome in patients requiring CVVH following acute renal failure. METHODS: One hundred seventeen patients between the age of 18 and 80 years were randomized to CVVH either with RF-bic (N = 61) or RF-lac (N = 56). Patients were treated with CVVH for five days or until either renal function was restored or the patient was removed from the study. Data were analyzed on day 5 or according to the "last observation carried forward" (LOCF) option. Adverse events were classified according to the WHO-Adverse Reaction Terminology system. RESULTS: Blood lactate levels were significantly lower and blood bicarbonate levels were significantly higher in patients treated with RF-bic than in those treated with RF-lac (lactate, 17.4 +/- 8.5 vs. 28.7 +/- 10.4 mg/dL, P < 0.05; bicarbonate, 23.7 +/- 0.4 vs. 21.8 +/- 0.5 mmol/L, P < 0. 01). The number of hypotensive crises was lower in RF-bic-treated patients than in RF-lac-treated patients (RF-bic 14 out of 61 patients, RF-lac in 29 out of 56 patients; 0.26 +/- 0.09 vs. 0.60 +/- 0.31 episodes per 24 h, P < 0.05). Nine out of 61 patients (15%) treated with RF-bic and 21 out of 56 patients (38%) treated with RF-lac developed cardiovascular events during CVVH therapy (P < 0. 01). A multiple regression analysis showed that the occurrence of cardiovascular events was dependent on replacement fluid and previous cardiovascular disease and not on age or blood pressure. Patients with cardiac failure died less frequently in the group treated with RF-bic (7 out of 24, 29%) than in the group treated with RF-lac (12 out of 21, 57%, P = 0.058). In patients with septic shock, lethality was comparable in both groups (RF-bic, 10 out of 27, 37%; RF-lac, 7 out of 20, 35%, P = NS). CONCLUSIONS: The results show that the administration of RF-bic solution was superior in normalizing acidosis of patients without the risk of alkalosis. The data also suggest that the use of RF-bic during CVVH reduces cardiovascular events in critically ill patients with acute renal failure, particularly those with previous cardiovascular disease or heart failure. PMID- 11012910 TI - Mortality caused by sepsis in patients with end-stage renal disease compared with the general population. AB - BACKGROUND: In the United States, infection is second to cardiovascular disease as the leading cause of death in patients with end-stage renal disease (ESRD), and septicemia accounts for more than 75% of this category. This increased susceptibility to infections is partly due to uremia, old age, and comorbid conditions. Although it is intuitive to believe that mortality caused by sepsis may be higher in patients with ESRD compared with the general population (GP), no such data are currently available. METHODS: We compared annual mortality rates caused by sepsis in patients with ESRD (U.S. Health Care Financing Administration 2746 death notification form) with those in the GP (death certificate). Data were abstracted from the U.S. Renal Data System (1994 through 1996 Special Data request) and the National Center for Health Statistics. Data were stratified by age, gender, race, and diabetes mellitus (DM). Sensitivity analyses were performed to account for potential limitations of the data sources. RESULTS: Overall, the annual percentage mortality secondary to sepsis was approximately 100- to 300-fold higher in dialysis patients and 20-fold higher in renal transplant recipients (RTRs) compared with the GP. Mortality caused by sepsis was higher among diabetic patients across all populations. After stratification for age, differences between groups decreased but retained their magnitude. These findings remained robust despite a wide range of sensitivity analyses. Indeed, mortality secondary to sepsis remained approximately 50-fold higher in dialysis patients compared with the GP, using multiple cause-of-death analyses; was approximately 50-fold higher in diabetic patients with ESRD compared with diabetic patients in the GP, when accounting for underreporting of DM on death certificates in the GP; and was approximately 30-fold higher in RTRs compared with the GP, when accounting for the incomplete ascertainment of cause of death among RTRs. Furthermore, despite assignment of primary cause-of-death to major organ infections in the GP, annual mortality secondary to sepsis remained 30- to 45-fold higher in the dialysis population. CONCLUSIONS: Patients with ESRD treated by dialysis have higher annual mortality rates caused by sepsis compared with the GP, even after stratification for age, race, and DM. Consequently, this patient population should be considered at high-risk for the development of lethal sepsis. PMID- 11012912 TI - Unrestricted pore area (A0/Deltax) is a better indicator of peritoneal membrane function than PET. AB - BACKGROUND: How to measure the peritoneal exchange in uremic patients treated with peritoneal dialysis (PD) is still a matter of controversy. Most clinics use the peritoneal equilibration test (PET), but from a theoretical point of view, it would be more appropriate to determine the "area" parameter, A0/Deltax. The latter reflects the total unrestricted pore area per centimeter diffusion distance and can be obtained by three-pore analysis using, for example, the PD capacity test (PDC). To evaluate the different estimates of peritoneal function, PET data and the A0/Deltax parameters were compared with the independently determined uptake of a small diffusible tracer, iohexol (molecular weight of 821 D), from the abdominal cavity to blood. METHODS: Fourteen patients on routine PD underwent determinations of PET and A0/Deltax using PDC. Within a month, the two hour uptake of iohexol (6 mg/mL) was also determined from the plasma iohexol concentration following abdominal filling. RESULTS: A strong correlation was found between the rate of iohexol plasma concentration increase (k30-120) and A0/Deltax (A0/Deltax = 76,300. k30-120 - 1.56; r2 = 0.799; N = 14) for the 2 L dwell, while the PET data were far less related to iohexol uptake (D/DPurea, r2 = 0.409; D/Pcreatinine, r2 = 0.436; and D/D0glucose, r2 = 0.015, respectively). CONCLUSION: The "area" parameter, A0/Deltax, is superior to the more widely used routine PET as an indicator of peritoneal membrane function. In addition, the concept of A0/Deltax has the virtue of supplying quantitative information about the peritoneal pathophysiology and physiology. PMID- 11012911 TI - Effects of lactate-buffered and lactate-free dialysate in CAVHD patients with and without liver dysfunction. AB - BACKGROUND: Continuous modalities of renal replacement deplete patients of bicarbonate, which is traditionally replaced indirectly by lactate in dialysate or replacement fluids. We have compared a new lactate-free dialysate (unbuffered dialysate with separate bicarbonate replacement of dialytic bicarbonate loss) with standard lactate-buffered dialysate in terms of acid-base control, lactate accumulation, and hemodynamic stability in patients undergoing continuous renal replacement therapy in an intensive care unit. METHODS: A nonrandomized crossover cohort study involving 54 patients with multi-organ failure (of whom 19 had significant hepatic dysfunction) was performed. All patients completed 24-hour continuous hemodiafiltration against both lactate-buffered and lactate-free dialysate. Arterial pH, blood gases, bicarbonate, and lactate, venous sodium, blood pressure, and inotrope requirements were measured before and at six hourly intervals during the first 24 hours of dialysis against each dialysate. RESULTS: Lactate-free dialysate provided more rapid control of acidosis than lactate buffered with less total administration of buffer than that given during the lactate-buffered period (total mmol bicarbonate vs. total mmol lactate + bicarbonate). Lactate accumulation was slight in both periods, but was higher during lactate-buffered continuous venovenous hemodiafiltration (CVVHD). The mean arterial pressure rose during lactate-free dialysis with decreased inotrope doses and fell during lactate-buffered dialysis with increased inotrope requirement. Results in patients with liver dysfunction were not significantly different from those without it. CONCLUSIONS: Over the time scale of 24 hours, lactate derived from continuous dialysis circuits is efficiently cleared from the blood of most patients with multi-organ failure, but with less effect on systemic acidosis than is produced by equivalent amounts of bicarbonate. PMID- 11012913 TI - Nitric oxide inhibits the formation of advanced glycation end products. AB - BACKGROUND: Advanced glycation end products (AGEs) are elevated in renal failure and have been implicated in the pathogenesis of several uremic complications. Their formation is closely associated with oxidative stress. The recent observation that nitric oxide (NO) has an antioxidant effect led us to examine the possible role of NO in the generation of AGEs. METHODS: We examined the effect of NO donors, 2, 2'-(hydroxynitrosohydrazono)bis-ethanamine (NOC18) and S nitroso-N-acetyl-DL-penicillamine (SNAP), on the in vitro formation of pentosidine, which was used as a surrogate marker for AGEs. Bovine serum albumin was incubated under air at 37 degrees C in a medium containing either several AGE precursors or uremic plasma. To elucidate further the mechanism of the NO effect on AGE formation, we examined the generation of free radicals and carbonyls in pentose-driven pentosidine formation. RESULTS: NO donors significantly inhibit the formation of pentosidine in a dose-dependent manner. The effect is abolished by the addition of a NO scavenging agent, 2-(4-carboxyphenyl)-4,4,5, 5 tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO). The inhibitory effect results from NO but not from the NO donor molecule. It is best explained by the ability of NO to scavenge carbon-centered radicals, hydroxyl radical, and carbonyl compounds. CONCLUSIONS: NO inhibits pentosidine formation by scavenging free radicals and by inhibiting carbonyl compound formation. NO might be implicated in the atherogenic and inflammatory effects of AGEs: Reduced NO production and increased oxidative stress associated with atherosclerotic lesions may accelerate AGE formation and, thus, exacerbate endothelial dysfunction and accelerate the development of atherosclerosis in uremia. PMID- 11012914 TI - Cyclosporine bone remodeling effect prevents steroid osteopenia after kidney transplantation. AB - BACKGROUND: It is well established that prednisone above 7.5 mg/day may induce osteopenia in association with decreased bone formation. In contrast, the effect of cyclosporine on bone remodeling and bone mineral density (BMD) is controversial. Multiple confounding factors explain this controversy, especially after renal transplantation. METHODS: Fifty-two renal transplanted patients never exposed to aluminum while on dialysis were selected because they had no rejection and no hypercalcemia for 24 months while being treated with low dose prednisone/cyclosporine A (daily dose at 10 mg and 4.8 mg/kg, respectively, beyond 3 months). Bone remodeling markers (BRMs; plasma osteocalcin, bone and total alkaline phosphatases for formation, and urinary pyridinolines for resorption) were sequentially measured together with plasma creatinine, intact parathyroid hormone (PTH) and 25 OH vitamin D and cyclosporine from day 0 to 24 months. BMD was measured at 3, 6, 12, and 24 months by quantitative computerized tomography (QCT) at the lumbar spine and by double-energy x-ray absorptiometry (DEXA) at this site, as well as at the femoral neck, radius shaft, and ultradistal (UD) radius. RESULTS: Plasma concentrations of creatinine, PTH, and 25 OH vitamin D initially decreased and stabilized beyond three months at 137 micromol/L, 1.5 the upper limit of normal (ULN) and 11 ng/mL, respectively. All BRM increased significantly above the ULN at six months and then decreased. The BMD Z score at three months was low at all sites measured by DEXA and QCT. Follow up measurements showed stability of absolute value and of Z score at all sites measured by DEXA. A comparison of the lumbar QCT Z score, which was available in 42 patients at 3 and 24 months, showed an increase in 28 and a decrease in 14, so that the increase for the whole group was significant (P < 0.04). Compared with patients with a decreased Z score, those with an increased Z score had significantly higher cyclosporine and lower prednisone dosages and a greater BRM increase at six months, whereas age, sex ratio, and plasma creatinine, PTH and 25 OH vitamin D were comparable and stable from months 3 through 24. The mean trough level of cyclosporine for the first six months was positively correlated to osteocalcin and total alkaline phosphatase increase at six months, and both bone formation and resorption marker increases were significantly correlated to the lumbar QCT Z score increase at 24 months. CONCLUSIONS: Combined low-dose prednisone and cyclosporine immunosuppression are associated with a stabilization of BMD measured at all sites with DEXA 3 to 24 months after renal transplantation and with a prevention of age-related loss of vertebral trabecular bone, as shown by the significant increase in lumbar spine QCT Z score. It is suggested that cyclosporine, together with the decrease of prednisone dosage but independent of renal function, PTH, and vitamin D status, contributes to a transient stimulation of bone remodeling at six months, which counterbalances the deleterious effect of prednisone on bone formation and BMD. PMID- 11012915 TI - Progressive adriamycin nephropathy in mice: sequence of histologic and immunohistochemical events. AB - BACKGROUND: As an experimental analogue of human focal glomerular sclerosis (FGS), adriamycin (ADR)-induced nephropathy is well-characterized in rats. Previously, this model has not been fully established in mice. The extension of this model to the mouse would be useful in the application of genetic and monoclonal antibody technology to characterize mechanisms of progressive renal disease. Herein, we describe a stable and reproducible murine model of chronic proteinuria induced by ADR. METHODS: Male BALB/c mice were intravenously injected with a single dose of ADR (10 to 11 mg/kg). Seven mice were sacrificed at weeks 1, 2, 4, and 6. Renal function, quantitative morphometric analysis, and electron microscopic studies were performed. Peripheral CD4+ and CD8+ T cells were evaluated using flow cytometric analysis of splenocytes. The leukocytic inflammatory pattern was analyzed by immunohistochemical examination. RESULTS: Overt proteinuria was observed from day 5 and remained significantly elevated throughout the study period. A focal increase in reabsorption droplets in tubular cells appeared at weeks 1 and 2, and numerous intraluminal casts were present after two weeks. Glomerular vacuolation and mild FGS appeared at week 4. At week 6, extensive focal and even global glomerular sclerosis, associated with moderate interstitial expansion and severe inflammation, were observed. A prominent macrophage infiltration appeared within both interstitium and glomeruli at week 2, followed by accumulation of both CD4+ and CD8+ T cells in interstitium but not glomeruli. There were almost no B lymphocytes seen at any time. CONCLUSION: This model should be useful in unraveling the pathogenesis of glomerular and interstitial inflammation and fibrosis in chronic proteinuric renal disease. PMID- 11012916 TI - Computational analysis of blood volume curves and risk of intradialytic morbid events in hemodialysis. AB - BACKGROUND: Blood volume (BV) curves have been used to prevent intradialytic morbid events (IMEs) caused by hypotensive episodes in hemodialysis treatment. However, no standardized parameter is available to describe BV dynamics and to enable online interference with ultrafiltration rates in unselected patients. Moreover, only time-dependent BV reduction and absolute hematocrit threshold, but not BV variability, have been suggested as markers of pending hypotension. The present study therefore deals with a computer-aided analysis of indices characterizing both BV reduction per time and BV variability in treatments of nonselected maintenance hemodialysis patients. METHODS: The methodology uses indices obtained by mathematical analysis of BV curves and was designed to potentially enable automatic interference with ultrafiltration. RESULTS: In 46 out of 380 treatments (12.1%), IMEs occurred. In these treatments, the indices for long- and short-term variability and slope of the curves were significantly lower than in treatments without IMEs. Moreover, the last 10 minutes before an IME were characterized by additionally decreased variability and slope. In a risk analysis of long-term variability and IMEs, we established an index below 16 to be associated with the highest risk of IMEs. CONCLUSIONS: Using these kind of index thresholds and online analysis of BV curves, automatic management of ultrafiltration by BV dynamics could be a promising concept to avoid intradialytic morbidity. PMID- 11012917 TI - Color doppler ultrasonography imaging to guide transluminal angioplasty of venous stenosis. AB - BACKGROUND: The objective of access surveillance is the early recognition of dysfunction in order to be able to correct the stenosis by angioplasty or surgery before access thrombosis occurs. The advent of color Doppler imaging has enabled studies of color Doppler ultrasonography (CDU) for the guidance of percutaneous transluminal angioplasty (PTA). The aim of the present study was to investigate whether color Doppler imaging alone can be safely and effectively used to diagnose vascular graft access stenoses and guide subsequent PTA. METHODS: Using the ultrasound velocity dilution method, we measured access blood flow (Qa) during the first hour of hemodialysis every month in patients with grafts as vascular access. When the decrease in Qa from the baseline value was 40% or more, CDU was performed and immediately followed by PTA in the presence of a stenosis of more than 50%. The Qa was then measured during the first dialysis after PTA and one month later. Repeated-measure analysis of variance was applied to evaluate the early and late (after one month) effect of PTA. RESULTS: Twelve PTAs were performed under CDU guidance in nine patients and led to the elimination of the stenosis or its reduction (two cases). The mean Qa was 809 +/- 263 mL/min at baseline, 468 +/- 153 before PTA, and 820 +/- 281 after PTA. The difference between the pre-PTA and post-PTA values was highly significant (P < 0.001), and the mean value after PTA was not different from baseline (P = 0.672). There were no relevant complications directly related to the procedure. CONCLUSIONS: The CDU procedure is effective for the diagnosis of vascular access stenosis and as a guide during the PTA procedure. It could improve stenosis screening by avoiding the risks of exposure to ionizing radiation and of adverse reactions to contrast media. PMID- 11012918 TI - Nitric oxide, advanced glycation end products, and uremia. PMID- 11012919 TI - Light from DARCness. PMID- 11012921 TI - Reply from the authors PMID- 11012920 TI - Renoprotection and blood pressure. PMID- 11012923 TI - In this issue PMID- 11012922 TI - Expanding the kidney donor pool: ethical and medical considerations. PMID- 11012924 TI - The Cambridge Conference: background. PMID- 11012925 TI - Combining marks, scores and grades. Reviewing common practices reveals some bad habits. PMID- 11012926 TI - Academic medicine In progress: reports of new approaches in medical education PMID- 11012927 TI - Marks, scores and grades: scaling and aggregating student assessment outcomes. AB - The term marks conflates the concepts of scores (raw test performance) and grades (level of performance). Neither scores nor grades represent interval scales, and therefore properly speaking arithmetic means should not be calculated during aggregation. The distributions of scores from a variety of kinds of assessment are considered, and ways of converting scores to grades are discussed. Methods of aggregation are also considered, and several strategies for implementing these via spreadsheets are made available. It is recommended that: 1 Scores should always be converted to grades before aggregation. The process of converting scores to grades requires both subject-specific skills, and familiarity with educational principles. 2 Whatever grade scale is used, it should be readily distinguishable from scores. 3 The median should be calculated as the measure of overall performance, not the arithmetic mean. 4 The interquartile range should be calculated as the measure of dispersion. 5 Students should be informed of both their score and grade for each assessment. 6 Where possible, assessment should report performance by individual assessment, not by a single aggregated mark. 7 When aggregation takes place, it should be possible to aggregate student performance by type of assessment as well as by academic subject. 8 Students who perform inconsistently should receive particular scrutiny during assessment. PMID- 11012928 TI - Development of a three-centre simultaneous objective structured clinical examination. AB - OBJECTIVE: To describe the development, organization, implementation and evaluation of a yearly multicentre, identical and simultaneous objective structured clinical examination (OSCE). SUBJECTS: All fifth-year medical students in a 6-year undergraduate medical programme. SETTING: The Christchurch, Dunedin and Wellington Schools of Medicine of the University of Otago, New Zealand. METHOD: One practice and two full 18-station OSCEs have been completed over 2 years, for up to 72 students per centre, in three centres. The process of development and logistics is described. Data are presented on validity, reliability and fairness. RESULTS: Face and content validity were established. Internal consistency was 0.83-0. 86 and interexaminer reliability, as assessed by the coefficient of correlation, averaged 0.78. Students rated the OSCE highly on relevance. Of the total variance in total OSCE marks, the schools contributed 6.9%, and the students 93.1%, in the first year. In the second year the schools contributed 6.2% and the students 93.8%. CONCLUSION: Implementation of a psychometrically sound, multicentre, simultaneous and identical OSCE is possible with a low level of interschool variation. PMID- 11012929 TI - Does studying for an objective structured clinical examination make a difference? AB - PURPOSE: This study examines the extent to which second-year medical students studied for an objective structured clinical examination (OSCE), how they studied, and the impact of self-reported studying on OSCE performance. METHOD: One class of 113 medical students completed an end-of-second-year OSCE, held on two consecutive evenings. The OSCE was comprised of eight stations, each of which was of 20 minutes' duration. The OSCE was formative: students received performance feedback but were not graded. Prior to the OSCE, students completed a brief survey regarding their preparation for the OSCE and their perceptions of confidence, anxiety and preparedness. Only 78 students returned surveys with names, comprising the data for these analyses. RESULTS: Mean studying time was 3.3 h, ranging from 0 to 19 h. Studying time was positively associated with age and negatively associated with basic science examination scores. The most study time was dedicated to reviewing the physical examination textbook, class notes and supplemental course readings. The breadth of study strategies increased as more time was spent in OSCE preparation. OSCE performance was related to study time and to achievement on pre-clinical basic science examinations. DISCUSSION: The students whose performance was above average seemed to be the talented students whose records indicated a history of academic success. The amount of time they reported for OSCE preparation was comparable to that reported by students with below average performance. It appears that prior academic performance rather than preparatory studying time is a better predictor of OSCE outcomes. PMID- 11012930 TI - The management of patient encounter time in a high-stakes assessment using standardized patients. AB - OBJECTIVES: The purpose of this study was to gather information regarding the appropriateness of the length of time allotted for candidates to complete the history taking and physical examination tasks in a high-stakes standardized patient (SP) assessment. DESIGN: Data were collected on actual time used by 1548 examinees for each of their 10 standardized patient encounters, for which a maximum of 15 minutes was allotted, but not required. SETTING: The Clinical Skills Assessment Center of the Educational Commission for Foreign Medical Graduates (ECFMG), Philadelphia, Pennsylvania, USA. SUBJECTS: Graduates of foreign medical schools who are seeking ECFMG certification. RESULTS: The average time spent with the standardized patient was 13.3 minutes, suggesting that the 15 minute time limit was sufficient. A positive correlation was found between data gathering scores and patient interview times. Candidates did tend to spend more time with SPs presenting with cases involving complex histories, as well as with cases of chronic conditions. CONCLUSIONS: Candidate time use varied as a function of type of clinical encounter, providing additional evidence of the content validity of the Clinical Skills Assessment. PMID- 11012931 TI - Introduction to the papers from the ninth cambridge conference on medical education PMID- 11012932 TI - Clinical teaching: maintaining an educational role for doctors in the new health care environment. AB - CONTEXT AND OBJECTIVES: Good clinical teaching is central to medical education but there is concern about maintaining this in contemporary, pressured health care environments. This paper aims to demonstrate that good clinical practice is at the heart of good clinical teaching. METHODS: Seven roles are used as a framework for analysing good clinical teaching. The roles are medical expert, communicator, collaborator, manager, advocate, scholar and professional. RESULTS: The analysis of clinical teaching and clinical practice demonstrates that they are closely linked. As experts, clinical teachers are involved in research, information retrieval and sharing of knowledge or teaching. Good communication with trainees, patients and colleagues defines teaching excellence. Clinicians can 'teach' collaboration by acting as role models and by encouraging learners to understand the responsibilities of other health professionals. As managers, clinicians can apply their skills to the effective management of learning resources. Similarly skills as advocates at the individual, community and population level can be passed on in educational encounters. The clinicians' responsibilities as scholars are most readily applied to teaching activities. Clinicians have clear roles in taking scholarly approaches to their practice and demonstrating them to others. CONCLUSION: Good clinical teaching is concerned with providing role models for good practice, making good practice visible and explaining it to trainees. This is the very basis of clinicians as professionals, the seventh role, and should be the foundation for the further development of clinicians as excellent clinical teachers. PMID- 11012933 TI - Effective supervision in clinical practice settings: a literature review. AB - CONTEXT: Clinical supervision has a vital role in postgraduate and, to some extent, undergraduate medical education. However it is probably the least investigated, discussed and developed aspect of clinical education. This large scale, interdisciplinary review of literature addressing supervision is the first from a medical education perspective. PURPOSE: To review the literature on effective supervision in practice settings in order to identify what is known about effective supervision. CONTENT: The empirical basis of the literature is discussed and the literature reviewed to identify understandings and definitions of supervision and its purpose; theoretical models of supervision; availability, structure and content of supervision; effective supervision; skills and qualities of effective supervisors; and supervisor training and its effectiveness. CONCLUSIONS: The evidence only partially answers our original questions and suggests others. The supervision relationship is probably the single most important factor for the effectiveness of supervision, more important than the supervisory methods used. Feedback is essential and must be clear. It is important that the trainee has some control over and input into the supervisory process. Finding sufficient time for supervision can be a problem. Trainee behaviours and attitudes towards supervision require more investigation; some behaviours are detrimental both to patient care and learning. Current supervisory practice in medicine has very little empirical or theoretical basis. This review demonstrates the need for more structured and methodologically sound programmes of research into supervision in practice settings so that detailed models of effective supervision can be developed and thereby inform practice. PMID- 11012934 TI - Strategic planning in medical education: enhancing the learning environment for students in clinical settings. AB - BACKGROUND: The 1999 Cambridge Conference was held in Northern Queensland, Australia, on the theme of clinical teaching and learning. It provided an opportunity for groups of academic medical educators to consider some of the challenges posed by recent changes to health care delivery and medical education across a number of countries. PURPOSE: This paper describes the issues raised by the practical challenges posed by the current environment and how they might be addressed in ways that could promote more effective learning in clinical settings. METHOD: A SWOT analysis is a tool that can help in forward planning by identifying the strengths, weaknesses, opportunities and threats presented by any situation. Our SWOT analysis was used to generate a list of items, from which we chose those most feasible and most likely to promote positive change. RESULTS: Twenty different issues were identified, with four of them chosen by consensus for further elaboration. The discussion gave rise to four main recommended strategies: ensuring that clinical teachers thoroughly understand the purpose and process of learning in clinical settings; equipping learners with 'survival skills'; making the best use of learning resources within different clinical environments and making judicious use of information technology to enhance learning efficiency. CONCLUSIONS: The four strategies were selected not only because of their inherent importance, but also because of their feasibility. Modest changes can motivate students to feel part of a clinical team and a 'community of practice' and enhance their capacity for self-regulated practice. PMID- 11012936 TI - In-training assessment - its potential in enhancing clinical teaching. AB - CONTEXT: In-training assessment (ITA) has established its place alongside formative and summative assessment at both the undergraduate and postgraduate level. In this paper the authors aimed to identify those characteristics of ITA that could enhance clinical teaching. METHODS: A literature review and discussions by an expert working group at the Ninth Cambridge Conference identified the aspects of ITA that could enhance clinical teaching. RESULTS: The features of ITA identified included defining the specific benefits to the learner, teacher and institution, and highlighting the patient as the context for ITA and clinical teaching. The 'mapping' of a learner's progress towards the clinical teaching objectives by using multiple assessments over time, by multiple observers in both a systematic and opportunistic way correlates with the incremental nature of reaching clinical competence. CONCLUSIONS: The importance of ITA based on both direct and indirect evidence of what the learner actually does in the real clinical setting is emphasized. Particular attention is given to addressing concerns in the more controversial areas of assessor training, ratings and documentation for ITA. Areas for future research are also identified. PMID- 11012935 TI - Patient-oriented learning: a review of the role of the patient in the education of medical students. AB - AIM: To explore the contribution patients can make to medical education from both theoretical and empirical perspectives, to describe a framework for reviewing and monitoring patient involvement in specific educational situations and to generate suggestions for further research. METHODS: Literature review. RESULTS: Direct contact with patients can be seen to play a crucial role in the development of clinical reasoning, communication skills, professional attitudes and empathy. It also motivates through promoting relevance and providing context. Few studies have explored this area, including effects on the patients themselves, although there are examples of good practice in promoting more active participation. CONCLUSION: The Cambridge framework is a tool for evaluating the involvement of patients in the educational process, which could be used by curriculum planners and teachers to review and monitor the extent to which patients are actively involved. Areas for further research include looking at the 'added value' of using real, as opposed to simulated, patients; more work on outcomes for patients (other than satisfaction); the role of real patients in assessment; and the strengths and weaknesses of different models of patient involvement. PMID- 11012937 TI - A review of the evaluation of clinical teaching: new perspectives and challenges. AB - PURPOSE: This article discusses the importance of the process of evaluation of clinical teaching for the individual teacher and for the programme. Measurement principles, including validity, reliability, efficiency and feasibility, and methods to evaluate clinical teaching are reviewed. CONTEXT: Evaluation is usually carried out from the perspective of the learner. This article broadens the evaluation to include the perspectives of the teacher, the patient and the institutional administrators and payers in the health care system and recommends evaluation strategies. RESULTS: Each perspective provides specific feedback on factors or attributes of the clinical teacher's performance in the domains of medical expert, professional, scholar, communicator, collaborator, patient advocate and manager. Teachers should be evaluated in all domains relevant to their teaching objectives; these include knowledge, clinical competence, teaching effectiveness and professional attributes. CONCLUSIONS AND IMPLICATIONS: Using this model of evaluation, a connection can be made between teaching and learning about all the expected roles of a physician. This can form the basis for systematic investigation into the relationship between the quality of teaching and the desired outcomes, the improvement of student learning and the achievement of better health care practice. It is suggested that the extent of effort and resources devoted to evaluation should be commensurate with the value assigned to the evaluation process and its outcomes. PMID- 11012938 TI - The accountability of clinical education: its definition and assessment. AB - BACKGROUND: Medical education is not exempt from increasing societal expectations of accountability. Competition for financial resources requires medical educators to demonstrate cost-effective educational practice; health care practitioners, the products of medical education programmes, must meet increasing standards of professionalism; the culture of evidence-based medicine demands an evaluation of the effect educational programmes have on health care and service delivery. Educators cannot demonstrate that graduates possess the required attributes, or that their programmes have the desired impact on health care without appropriate assessment tools and measures of outcome. OBJECTIVE: To determine to what extent currently available assessment approaches can measure potentially relevant medical education outcomes addressing practitioner performance, health care delivery and population health, in order to highlight areas in need of research and development. METHODS: Illustrative publications about desirable professional behaviour were synthesized to obtain examples of required competencies and health outcomes. A MEDLINE search for available assessment tools and measures of health outcome was performed. RESULTS: There are extensive tools for assessing clinical skills and knowledge. Some work has been done on the use of professional judgement for assessing professional behaviours; scholarship; and multiprofessional team working; but much more is needed. Very little literature exists on assessing group attributes of professionals, such as clinical governance, evidence-based practice and workforce allocation, and even less on examining individual patient or population health indices. CONCLUSIONS: The challenge facing medical educators is to develop new tools, many of which will rely on professional judgement, for assessing these broader competencies and outcomes. PMID- 11012939 TI - Graeme catto PMID- 11012940 TI - Residual strain in the gastrointestinal tract: a new concept. PMID- 11012941 TI - Human postprandial gastric emptying of indigestible solids can occur unrelated to antral phase III. AB - According to animal experiments, postprandial gastric emptying of indigestible solids is mainly related to the antral phase III activity of the migrating motor complex. Gastric emptying of indigestible solids in humans has not been directly correlated to pressure recordings. The aim of the present study was to investigate the postprandial emptying pattern of indigestible solids in humans and its relation to fed and fasted antral motility. Ten healthy volunteers participated. After an overnight fast they had a standard breakfast. Two sizes of radiopaque markers (ROMs) were given with the test meal; ten cubes each of side measurement 1.5 mm and 3 mm, respectively. Emptying of the ROMs from the stomach was followed by fluoroscopy with simultaneous antral manometry. In six of the subjects, fasting antral manometry was performed on one day and on another day, the emptying of 7 mm cylindrical particles together with 3 mm cubes, in the absence of a gastric tube was recorded. All ROMs were emptied within 5 h (range 1.5-4.5 h). In all subjects, the smaller particles (1.5 mm) showed a slight, insignificant tendency to move from the stomach more rapidly than the larger (3 mm) particles. None of the subjects had an antral phase III before all ROMs were emptied from the stomach. Instead, the typical irregular postprandial pressure activity was present in all subjects until the emptying was completed. Furthermore, the highest postprandial motility index during the emptying study was far below the motility index during phase III, but comparable to the motility index during late phase II. Emptying of the 7 mm particles occurred significantly more slowly at 1.5-2.5 h, but otherwise was similar to the emptying of the smaller particles. There was no difference between emptying of the 3 mm cubes with or without the presence of the tube. Contrary to common opinion, gastric emptying of indigestible solids after a meal can occur unrelated to the antral phase III, at least up to a particle size of 3 mm and perhaps even 7 mm. These findings are of great importance for the evaluation of gastric emptying of indigestible solids, including the pharmacodynamics of orally administered drugs. PMID- 11012942 TI - Acute intraduodenal bile salt depletion leads to strong gallbladder contraction, altered antroduodenal motility and high plasma motilin levels in humans. AB - Cholecystokinin is the main hormone involved in postprandial gallbladder contraction. There is also considerable gallbladder contraction in the fasting state, associated with phase III of the gastrointestinal migrating motor complex and release of the hormone motilin. It has been proposed that intraduodenal bile salts exert a negative-feedback control on postprandial cholecystokinin release and resulting gallbladder contraction. We wanted to elucidate whether a similar control mechanism on gallbladder contraction exists in the fasting state. We therefore performed gallbladder ultrasonography and 24-h antroduodenal motility registrations and determined plasma cholecystokinin and motilin levels in six healthy subjects before and after acute (4 g) and chronic (8 days; 8 g day(-1)) oral cholestyramine. Acute cholestyramine strongly decreased gallbladder volumes and increased motilin without changed cholecystokinin levels. There was a negative relationship between gallbladder volumes and plasma motilin levels. Although there was a persistent fasting pattern of antroduodenal motility, its cycle length was increased (P < 0.03) with markedly longer phase II (P < 0. 005). Fasting gallbladder volumes 24 h later were still strongly decreased but gradually increased to pretreatment levels. Before and after 8 days cholestyramine, interdigestive and postprandial gallbladder emptying, intestinal migrating motor complex and hormone levels did not differ. We conclude that acute (but not chronic) intraduodenal bile salt depletion with cholestyramine affects gallbladder and antroduodenal motility, possibly partly related to motilin release. PMID- 11012943 TI - Schistosoma mansoni infection causing diffuse enteric inflammation and damage of the enteric nervous system in the mouse small intestine. AB - Schistosomiasis mansoni is a major health problem, mainly occurring in developing countries. A large proportion of infected individuals suffers from motility related gastrointestinal problems. In the present study, the diffuse inflammatory response in the small bowel wall, as compared to the egg-induced granulomatous inflammation, was investigated. For this purpose, OF1 mice infected with Schistosoma mansoni 8-16 weeks prior to the experiment, and uninfected control mice were studied. The ileum showed both a diffuse mucosal inflammation as well as a granulomatous reaction. The diffuse mucosal inflammation caused an increase in the thickness of the mucosa, with blunting of the villi. A significant, transient increase of thickness of the muscularis propria after 12 weeks of infection was noted. There was an infection-related mast cell infiltrate in the muscularis propria, consisting of formalin fixation-insensitive connective tissue mast cells. Ganglionitis of the myenteric plexus was noted. Rarely, ganglia of the myenteric plexus contained apoptotic cells. A general pharmacological set of experiments showed a significant increase in intestinal contractility, both to exogenously administered, as well as to endogenously released neurotransmitters. Our results demonstrate that S. mansoni infection in the mouse ileum leads to diffuse specific enteric inflammation that is associated with an enhanced response to contractile agents. PMID- 11012944 TI - Endomorphin-1 and -2, endogenous ligands for the mu-opioid receptor, inhibit striated and smooth muscle contraction in the rat oesophagus. AB - Recently, morphological evidence for an interaction of autonomic nerve fibres and extrinsic motor innervation of the rat oesophagus has emerged. The aim of the present study was to investigate the possible influence of endogenous and exogenous opioids on rat oesophageal smooth and striated muscle function in vitro. The entire oesophagus (excluding the lower oesophageal sphincter) with both Nervi (Nn) vagi, including the Nn recurrentes, was dissected and placed in an organ bath (100 mL, 37 degrees) with oxygenated Krebs-Ringer buffer. Contractile activity was measured in a longitudinal direction with a force transducer. Both Nn vagi were placed on a bipolar platinum electrode 2 cm distant from the oesophagus. Vagal stimulation (VS), applied for 1 s (40 V, 0.5 ms, 20 Hz) resulted in a biphasic contractile response that was completely blocked by 10(-6) M tetrodotoxin. The first part consisted of a tetanic striated muscle contraction, as it was abolished by tubocurarine (10(-5) M, n=5) but unaffected by atropine (10(-6) M, n=3) or hexamethonium (10(-4) M, n=4). In contrast, the second part was completely inhibited by hexamethonium (10(-4) M) and atropine (10(-6)M), whereas tubocurarine (10(-5) M) showed no influence, indicating a stimulation of preganglionic nerve fibres supplying oesophageal smooth muscle (muscularis mucosae) via relays in myenteric ganglia. In order to characterize opioid influence on the oesophageal striated and smooth muscle contractility, the following experiments were carried out. 10(-6) M endomorphin-1 and -2, endogenous mu-opioid-receptor agonists, reduced the contractile response of the striated (EM 2, -25.1+/-5.3%; n=16), and the smooth muscle (EM-2, -81.9+/-3.3%; n=11). Both effects were reversible by the opioid receptor antagonist naloxone (10(-6) M) and therefore, mediated via opioid receptors. Neither SNC-80, an agonist on the delta opioid-receptor, U-69593, an agonist on the kappa-opioid-receptor, nor nociceptin, an agonist at the ORL1 (opioid receptor-like) receptor, had a significant effect on the striated muscle contraction. In contrast to SNC-80, U 69593 and nociceptin inhibited smooth muscle contraction but this relaxation could not be antagonized by naloxone. None of the opioid receptor antagonists used had an effect on basal tonus or muscle contraction following VS. Our data provide evidence for an autonomic modulation of vagal motor innervation of the striated and smooth oesophageal muscle. Endomorphin-1 and -2, both selective mu opioid receptor agonists, cause an inhibition of striated and smooth muscle response which is reversible by naloxone, an opioid receptor antagonist. The location of the mu-opioid receptor still has to be established. PMID- 11012945 TI - Increased mast cells in the irritable bowel syndrome. AB - Mast cells (MC) release potent mediators which alter enteric nerve and smooth muscle function and may play a role in the pathogenesis of the irritable bowel syndrome (IBS). The aim of this study was to determine if MC were increased in the colon of IBS patients compared to controls. Biopsy specimens were obtained from the caecum, ascending colon, descending colon and rectum of 28 patients: 14 IBS (Rome criteria); seven normal; and seven inflammatory controls. Tissue was stained immunohistochemically using a monoclonal mouse antibody for human mast cell tryptase (AA1). Tissue area occupied by tryptase-positive MC (volume density of mast cells) was quantified by image analysis. The number of plasma cells, lymphocytes, eosinophils, neutrophils and macrophages were each graded semiquantitatively (0-4) in haematoxylin and eosin stained sections. Mast cell volume density was significantly (P < 0.05) higher in IBS (0.91 +/- 0.18; CI 0.79; 1.0) than normal controls (0.55 +/- 0.14; CI 0.40; 0.69) in the caecum but not at other sites. Apart from MC, there was no evidence of increased cellular infiltrate in the IBS group. MC were significantly increased in the caecum of IBS patients compared to controls. The multiple effects of the intestinal mast cell alone, or as a participant of a persistent inflammatory response, may be fundamental to the pathogenesis of IBS. PMID- 11012946 TI - Involvement of tachykinin NK2 receptors in the modulation of spontaneous motility in rat proximal colon. AB - The role of endogenous tachykinins and the mechanisms whereby they act on NK2 receptors, modulating spontaneous motility, were investigated in rat isolated proximal colon. The mechanical activity was detected as changes in intraluminal pressure. The NK2 receptor antagonist, MEN 10627, produced a concentration dependent reduction of the contraction amplitude. [beta-Ala8]-neurokinin A(4-10), an NK2 receptor agonist, and [Sar9, Met(O2)11]-Substance P ([Sar9, Met(O2)11] SP), an NK1 receptor agonist, induced a concentration-dependent contractile response, characterized by an increase in basal tone with superimposed phasic contractions. MEN 10627 antagonized the response to [beta-Ala8]-neurokinin A(4 10), without affecting that to [Sar9, Met(O2)11]-SP. Tetrodotoxin (TTX), hexamethonium and Nomega-nitro-L-arginine methyl ester (L-NAME) significantly reduced the response to MEN 10627. The NK3 receptor agonist, senktide, was able to activate the nitrergic inhibitory pathway, as it induced a TTX-and L-NAME sensitive inhibitory effect. [beta-Ala8]-neurokinin A(4-10) was able to antagonize the inhibitory response to senktide. These findings suggest that tachykinins acting on NK2 receptors play a role in the modulation of the spontaneous mechanical activity. The mechanism of this action would be, in part, acting directly on the smooth muscle cells, and, in part neurogenic, sustained by nicotinic inputs, and possibly due to inhibition of NO tonic release. PMID- 11012948 TI - How much do we really know about postobstructive pulmonary oedema? PMID- 11012947 TI - The spatial behaviour of spike patches in the feline gastroduodenal junction in vitro. AB - In the isolated feline gastroduodenal region, the spatial propagation of slow waves and of individual spikes was reconstructed. Recordings were performed simultaneously from 240 extracellular electrodes positioned on the serosal surface across the junction. Results from nine experiments (22 slow waves) showed that the slow wave never propagated across the gastroduodenal region and that this block was due to the presence of a zone of quiescence caudal to the pylorus. In contrast, spikes (n=155) were able to propagate into the quiescent zone, either from the antrum (15.4%) or from the duodenum (34.0%) and occasionally, were able to propagate from one organ to the other (10.9%). However, in all cases, spike conduction was self-limited and activated a local area termed a 'patch'. The length of the patches located in the gastroduodenal region was significantly longer than in the rest of the duodenum (20.2 mm +/- 9. 7 vs. 9.5 mm +/- 3.2; P < 0.001) indicating a possible enhancement of spike propagation in this region. In conclusion, in spite of the total conduction block for slow waves, individual spikes are able to propagate across the gastroduodenal region, albeit in self-limited areas or 'patches'. These spike patches could form the building blocks for gastroduodenal coordination. PMID- 11012949 TI - The perioperative management of children with phaeochromocytoma. AB - The safe anaesthetic management of a child with a phaeochromocytoma requires an understanding of the pathophysiology of the disease, together with a thorough knowledge of its pharmacology, in order to avoid or minimize the potentially harmful cardiovascular changes that may occur during anaesthesia. Although there is a considerable amount of information on the management of the adult with phaeochromocytoma, much less has been written concerning children with the disease. Children differ significantly from adults in the incidence, location, presentation and management of this condition and these differences are discussed here together with some of the more controversial issues of management. PMID- 11012950 TI - Preoperative psychological preparation for children undergoing ENT operations: a comparison of two methods. AB - A psychological preparation programme was developed for outpatient surgery in children. The purpose of this study was to determine if the programme could increase retrieval of information and reduce anxiety prior to ENT surgery. After ethical committee approval, 160 children and their parents were included. Eighty children (group 1) received conventional verbal information from an ENT nurse, and another 80 children (group 2) received specific information, including role play, from a nurse anaesthetist at a preadmission visit. Children's and parents' experience of premedication, operation theatre (OR), i.v.-needle insertion and induction of anaesthesia were evaluated from a self-rating questionnaire. The questionnaire included ratings for anxiety and satisfaction with information and care. The results indicate a clear improvement of the preoperative acquisition of knowledge in all age groups. When it comes to alleviation of fear, a positive effect of the preparation programme was noticed, especially among the younger children (< 5 years), while preoperative anxiety overall was a significantly smaller problem among the older children. The effects of the programme were also related to previous experience of anaesthesia and most beneficial among young children with such experience. Overall, the most negative procedure reported by the children was the i.m. injection for premedication (a routine which was abandoned as a result of the study), followed by the insertion of the i.v. needle. The parents experienced watching their child fall asleep during induction of anaesthesia as most negative, followed by the insertion of the i.v.-needle. Parents also reported more satisfaction and less anxiety after having received specific information and preparation preoperatively. It was concluded that this preoperative preparation programme is useful in all age groups with regard to information, while alleviation of anxiety and fear was seen mainly among the younger children with previous experience of anaesthesia. PMID- 11012951 TI - Parental presence during anaesthesia induction for outpatient surgery of the infant. AB - We conducted a randomized controlled trial of parental presence during anaesthesia induction for outpatient surgery in 73 infants (aged 1-12 months). Effects of parental presence on infant and parental outcomes, including anxiety, health care attitudes and satisfaction with the anaesthesia and surgery experience were evaluated. Results demonstrated that parental presence had no impact on infant behavioural distress during induction. In addition, parents who were present demonstrated comparable anxiety levels and health care attitudes before and after surgery, as well as comparable levels of satisfaction with the surgical experience compared to parents who were absent during induction. We discuss reasons for the lack of treatment effects from parental presence, and new directions for future research to identify subgroups of children who may most benefit from the opportunity to have parents involved in the perioperative period. PMID- 11012952 TI - The time-course of action and recovery of rocuronium 0.3 mg x kg(-1) in infants and children during halothane anaesthesia measured with acceleromyography. AB - This study compares the time-course of action of neuromuscular paralysis after 0.3 mg x kg(-1) of rocuronium during nitrous oxide-halothane anaesthesia in children of three different age groups. With appropriate approval and informed consent from the parents, 51 children, ASA I-II, scheduled for elective surgery requiring muscle relaxation, were studied. The children were assigned to three groups according to age: group 1, 0-6 months; group 2, 6-24 months; and group 3, > 24 months of age. Induction of anaesthesia and tracheal intubation were performed under halothane anaesthesia. Acceleromyography of the thumb was recorded after supramaximal transcutaneous ulnar nerve stimulation using train-of four (TOF) stimulation. Rocuronium 0.3 mg x kg(-1) was given as a rapid i.v. bolus prior to surgical incision. The onset time (time to max effect) and the maximal depth of the block, the time to recovery of the first twitch (T1) to 25% and 75% of its baseline, the recovery index (RI), and the time to recovery of the TOF ratio to 70% after the end of injection of rocuronium were all measured. The mean (SD) age of the children in groups 1, 2, and 3 was 3.1 (1.6), 12.6 (3.7), and 63 (46) months, respectively. The onset time of rocuronium was 47 (12), 83 (42) and 94 (12) s, respectively, in groups 1, 2, and 3 (P<0.05 group 1 vs. 2 and 3). One hundred percent block was achieved in 18/19 patients in group 1, 12/14 in group 2 and 6/18 in group 3. The times to 25% and 75% recovery of T1 and the time for recovery of the TOF ratio to 70% were all significantly longer in groups 1 and 2 compared to group 3. Group 1 and 2 showed no significant differences in recovery times. The RI was significantly prolonged in group 1 versus 3. The authors conclude that rocuronium 0.3 mg x kg(-1) during halothane anaesthesia causes more neuromuscular depression and has a longer duration of action in infants than in children older than 2 years. PMID- 11012953 TI - Comparison of ropivacaine 0.1% and 0.2% with bupivacaine 0.2% for single-shot caudal anaesthesia in children. AB - We compared analgesic efficacy and degree of motor block induced by ropivacaine 0.1% (R 0.1) and 0.2% (R 0.2) vs. bupivacaine 0.2% (B 0. 2) after caudal anaesthesia in children. Total and free plasma concentrations were measured after caudal injection. Duration of caudal analgesia (median/range) was significantly shorter in group R 0.1 (1.7 h/0.2-6 h) than in group R 0.2 (4.5 h/1.7-6 h) or group B 0. 2 (4 h/1-6 h) (P<0.05). Motor block in the first 2 h postoperatively was significantly less for both ropivacaine groups compared with bupivacaine (P<0.05). Peak plasma concentrations after ropivacaine 0.2% were higher and protein binding lower than after bupivacaine 0.2% (P<0.05). We conclude that caudal analgesia with ropivacaine 0.1% is less effective and of shorter duration than that of ropivacaine 0.2%, whereas ropivacaine 0.2% provides pain relief similar to bupivacaine 0.2%. Motor block in the early postoperative period is less with ropivacaine than with bupivacaine. PMID- 11012954 TI - Comparison of three techniques for internal jugular vein cannulation in infants. AB - Central venous cannulation allows accurate monitoring of right atrial pressure and infusion of drugs during the anaesthetic management of infants undergoing cardiopulmonary bypass. In this prospective, randomized study, we compared the success and speed of cannulation of the internal jugular vein in 45 infants weighing less than 10 kg using three modes of identification: auditory signals from internal ultrasound (SmartNeedle, SM), external ultrasound imaging (Imaging Method, IM) and the traditional palpation of the carotid pulsation and other landmarks (Landmarks Method, LM). The cannulation time, number of attempts with LM and SM techniques were greater than those with IM technique. The incidence of carotid artery puncture and the success rate were not significantly different among the three groups. In infants, a method based on visual ultrasound identification (IM) of the internal jugular vein is more precise and efficient than methods based on auditory (SM) and tactile perception (LM). PMID- 11012955 TI - Anaesthesia with midazolam and S-(+)-ketamine in spontaneously breathing paediatric patients during magnetic resonance imaging. AB - We evaluated safety and efficacy of a sedation technique based on rectal and intravenous S-(+)-ketamine and midazolam to achieve immobilization during Magnetic Resonance Imaging (MRI). Thirty-four paediatric patients were randomly assigned to undergo either the sedation protocol (study group) or general anaesthesia (control group). Imaging was successfully completed in all children. Children in the study group received a rectal bolus (0.5 mg x kg(-1) midazolam and 5 mg x kg(-1) S-(+)-ketamine) and required additional i.v. supplementation (20+/-10 microg x kg(-1) x min(-1) S-(+)-ketamine and 4+/-2 microg x kg(-1) x min(-1) midazolam), spontaneous ventilation was maintained. Transient desaturation occurred once during sedation and four times in the control group (P=0.34). PECO2 was 5.3+/-0.5 kPa (40+/-4 mm Hg) in the study group and 4.1+/-0.6 kPa (31+/-5 mm Hg) in the control group (P<0.001). Induction and discharge times were shorter in the study group (P<0.001), recovery times did not differ significantly between the groups. Our study confirms that a combination of rectal and supplemental intravenous S-(+)-ketamine plus midazolam is a safe and useful alternative to general anaesthesia for MRI in selected paediatric patients. PMID- 11012956 TI - Analgesic efficacy of ketorolac 0.5% ophthalmic solution (Accular) in paediatric strabismus surgery. AB - This prospective double-blind study was designed to assess the analgesic efficacy of ketorolac 0.5% ophthalmic solution compared with placebo in 30 healthy children undergoing extraocular muscle recession for correction of strabismus. After paracetamol 20 mg.kg-1 preoperatively, a standard anaesthetic was given. There were no significant differences in Children's Hospital of Eastern Ontario Pain Scale (CHEOPS) and faces pain scale (FPS) scores, requirement for supplementary analgesia or in postoperative vomiting between the two groups over the following 24 h. This study did not demonstrate improved postoperative analgesia when topical ketorolac eye drops were given in addition to paracetamol. This observed lack of efficacy may reflect difficulties in the use of CHEOPS and FPS in this age group with this pain model. PMID- 11012957 TI - Percutaneous central venous catheterization in small infants: axillary block can facilitate the insertion rate. AB - Central venous cannulation through a peripheral vein is the technique of choice in awake nonsedated critically ill infants. Such a technique has a high failure rate. We undertook a retrospective study to determine whether a brachial plexus block performed via the axillary approach could improve the success rate for the insertion of a central venous catheter from a peripheral vein of the upper limb in small infants. Data from 128 infants, submitted or not submitted to the axillary block, were analysed. The failure rate for insertion of the central venous catheter was 27% in the group without the use of the axillary block and 9% with the axillary block (P<0.05). The use of brachial plexus block via the axillary route, although evaluated retrospectively, improves the success rate for the insertion of small diameter central venous silicon catheter from a peripheral vein of the upper limb in small infants. PMID- 11012959 TI - Parental opinions regarding the route of administration of analgesic medication in children. AB - Parents were asked about the acceptability of different routes of administration of analgesia for their children. Their opinions were also sought regarding the need for consent and of the sources and quality of information provided to them perioperatively. Questionnaires before and after surgery were administered to 150 consecutively recruited parents. The majority (58%) of parents considered the rectal route to be the most unpleasant way of giving medication, compared to 19% for intramuscular and 11% and 9% for intravenous and oral, respectively. Over 70% thought additional consent, either verbal or written, is not necessary for administering medication by any route. The best information was received verbally in the opinion of 90%. Contrary to expectations, the rectal route was the most unpopular of all in this study. Parents' limited knowledge and experience may be important determining factors. Despite their negative opinions, parents did not consider separate consent necessary for the rectal or any other route. PMID- 11012958 TI - The feasibility of pain treatment at home after adenoidectomy with ketoprofen tablets in small children. AB - In this study, we investigated the feasibility of pain treatment using ketoprofen 25 mg tablets (5 mg x kg(-1) x day(-1)) at home in children after daycase adenoidectomy. We also determined the adverse events and the incidence of postoperative bleeding during the first week after surgery. Initially, we studied 611 children aged 1-9 years. The study design was prospective, longitudinal, and open. The final data consisted of 555 (91%) children, and 522 children who received ketoprofen at home. The parents administered four (1-10, median with 10th and 90th percentiles) ketoprofen tablets to their children during the first week. A total of 20% of the parents experienced problems in administering tablets, and problems were three times more common in children under 48 months compared to older children. The main problems were swallowing difficulties and the unpleasant taste of the tablet. Neither serious adverse events, nor clinically significant bleeding occurred. Ketoprofen at the dose of 5 mg x kg(-1) x day(-1) proved to be a safe analgesic in children for short-term use after adenoidectomy. PMID- 11012960 TI - Anaesthetic management of children with congenital insensitivity to pain with anhidrosis. AB - Congenital insensitivity to pain with anhidrosis (CIPA) is a rare sensory neuropathy, which affects patients' pain sensation and thermoregulation. There are several issues to consider when planning anaesthesia for those with this congenital disorder. Over a 20-year period, six patients with CIPA underwent 20 surgical procedures under general anaesthesia in our institution. We analysed our experience with these patients retrospectively. We conclude that patients with CIPA are able to undergo surgical procedures under general anaesthesia without major problems. PMID- 11012961 TI - The topics of international publications on paediatric anaesthesia from 1993 to 1998. AB - A comprehensive compilation of the current international literature on paediatric anaesthesia is still lacking. It was the aim of this study to identify all publications with a focus on paediatric anaesthesia, and to determine the spectrum of topics, as well as the publication type and language for the period between 1993 and 1998. All articles published in 12 major anaesthesia journals were evaluated and, additionally, a computerized, Internet-based Medline-search was performed using selected keywords. The analysis was limited to original articles, case reports, reviews and editorials. For the period between 1993 and 1998, a total of 2259 (377 per year) publications on paediatric anaesthesia were identified in 295 different journals, the majority of which were on the topic of 'providing anaesthesia' in children (n=1424, 63.0%). In contrast, publications on, for example, 'postanaesthesia care' (6. 3%), and 'organizational aspects of paediatric anaesthesia' (2.2%) were rare. Most articles were written in English (85.1%), and more than 50% reported original data (57.1%). Our results suggest that several topics may be of interest for future research and communication in the field of paediatric anaesthesia and new results should be published in English to reach a large international readership. PMID- 11012962 TI - Protein concentration in pulmonary oedema fluid for negative pressure pulmonary oedema in children. AB - We report a case of acute pulmonary oedema following upper airway obstruction in a 1-year-old patient. We discuss the pathophysiology of this infrequent complication of upper airway obstruction and the interest of alveolar protein/blood protein ratio measurement to determine the mechanism of pulmonary oedema. PMID- 11012963 TI - Pseudopolyp of the right laryngeal ventricle following atraumatic intubation: a diagnostic dilemma. AB - The traumatic effects of tracheal intubation are well recognized. Whilst these usually follow prolonged intubation, acute lesions have been described, usually following a traumatic intubation technique. We present a case of acute, localized swelling of the right laryngeal ventricle which followed an entirely atraumatic intubation by an experienced paediatric anaesthetist. The lesion was not present at the time of intubation, but developed subsequently during the surgical procedure. Although previously observed at our institution, such lesions have not been described in the literature. The potential for a diagnostic pitfall, by mistaking the lesion for a laryngeal cyst or nodule, is discussed. PMID- 11012964 TI - Inadvertent administration of intravenous ropivacaine in a child. AB - Following abdominal surgery with inhalation anaesthesia and epidural ropivacaine analgesia, inadvertent intravenous (i.v.) administration of ropivacaine occurred in a 1-year-old boy. The child spent 75 min in the postanaesthesia care unit and was transferred to the paediatric intensive care unit. Two hours after transfer, it was noted that the epidural tubing was connected to the peripheral i.v. line. The child remained awake, vital signs were stable, and his oxygen saturation ranged from 96-98% on room air. The epidural catheter was removed. He did not require further pain relief for the next 10 h. PMID- 11012965 TI - Positive test dose in a neonate with a caudally placed epidural catheter. AB - We present a case of a neonate with an epidural catheter placed via the caudal route after induction of general anaesthesia in whom the test doses of epinephrine-containing local anaesthetic was positive on two occasions. Remarkable tachycardia was noted after each of two separate injections through the catheter. Blood was never aspirated from the catheter and placement was without difficulty. After the catheter was removed, blood was noted at the tip. PMID- 11012966 TI - Train of four fade in a child with stiff baby syndrome. AB - A case is described of a child with stiff baby syndrome who underwent open reduction and femoral shortening of congenital dislocated hip under general anaesthesia. Neuromuscular function was measured electromyographically and demonstrated a great degree of train of four fade (57%) after sevoflurane inhalational induction of anaesthesia. The response to suxamethonium (2 mg x kg( 1)) was normal. The neuromuscular response to volatile anaesthetic agents and suxamethonium may be abnormal in these children with stiff baby syndrome and intraoperative neuromuscular monitoring is recommended. PMID- 11012967 TI - Anaesthetic management of erythropoietic protoporphyria. PMID- 11012968 TI - Novel use of neonatal cuffed tracheal tube to occlude tracheo-oesophageal fistula. PMID- 11012969 TI - Authors' reply PMID- 11012970 TI - Thumbs up for a new epidural catheter clamp! PMID- 11012971 TI - Horner syndrome due to thoracic epidural analgesia in children. PMID- 11012972 TI - Spinal anaesthesia in infants and children. PMID- 11012973 TI - Echinococcus granulosus: assays for hydatid immunoregulatory factors using established lymphoid cell lines. AB - Mitosis, mitochondrial metabolic rate and proliferation were measured in established lymphoid cell lines exposed to chromatographic fractions of equine Echinococcus granulosus hydatid fluid. In several cell lines, one or more of the three parameters were modified by the exposure. As an assay for potential immunoregulatory activity, the method was simple and repeatable. The following novel observations were made: (1) Mitotic reaction was found among lines of T cell, B-cell and macrophage origin; (2) mitosis was accompanied by proliferation in the B-cell lines, B9 and A20, and in the macrophage lines, HL-60 and P388d. With mitotically responsive T-cells, proliferation was slight in CTLL-2 and absent in D10, implying cell-cycle modification; (3) mitotic responsiveness tended to occur in cell lines with mature characteristics; (4) among cytokine dependent cell lines, hydatid fluid FPLC fraction 1 mimicked IL-1 and several fractions mimicked IL-2 and IL-6 in the maintenance of mitosis; and (5) there was significant statistical interaction between the influences of mammalian cytokines and hydatid fluid fractions, implying that the propensity of antigenically unprimed lymphoid cells to be regulated by E. granulosus is conditioned by cytokine activity. PMID- 11012974 TI - Interleukin-5 deficient mice exhibit impaired host defence against challenge Trichinella spiralis infections. AB - Enteric nematode infections are characterized by both peripheral and tissue eosinophilia. The cytokine interleukin (IL)-5 is considered a critical factor in the proliferation and recruitment of eosinophils, however, studies suggest it plays little role in host defence, at least during primary Trichinella spiralis infections. Less is known concerning its role in host defence or in the inflammatory response that develops against challenge infections with the same parasite. We examined these questions by infecting IL-5 deficient and wild-type mice, with T. spiralis parasites. Both strains expelled the primary infection by day 21. Forty days after the primary infection, we challenged the mice with a second T. spiralis infection and counted tissue eosinophils and worms in the intestine. While wild-type mice developed a large tissue eosinophilia, IL-5 deficient mice showed little increase in eosinophil numbers within the intestine. Throughout the challenge infection, significantly larger worm burdens were recovered from IL-5 deficient mice, and worm expulsion was also significantly slower (day 21) compared to wild-type mice (day 14). Thus, unlike in a primary infection, IL-5 is not only essential for the onset of intestinal eosinophilia, but also makes a significant contribution to enteric host defence during challenge T. spiralis infections. PMID- 11012975 TI - The roles of the glycosylphosphatidylinositol anchor on the production and immunogenicity of recombinant ookinete surface antigen Pbs21 of Plasmodium berghei when prepared in a baculovirus expression system. AB - Malarial ookinetes express an immunodominant surface protein (P28) that is a priority candidate for the development of transmission-blocking vaccines. The full length P28 gene from Plasmodium berghei [Pbs21(1-213)] and a deletion construct [Pbs21(1-188)] encoding a protein that lacks the 25 C-terminal amino acids, including the glycosylphosphatidylinositol (GPI) anchor signal, were expressed in insect cells using baculovirus vectors. Pbs21(1-213) protein is strongly hydrophobic, found in the cytoplasm and on the surface of Spodoptera Sf21 cells, and in the culture medium. Pbs21(1-188) protein was largely found in the aqueous phase of the medium and in the cytoplasm of Sf21 cells, but was not detected on the cell surface. The presence of 25 C-terminal amino acids is therefore critical to the attachment of recombinant Pbs21 to the parasite plasma membrane. Mice were immunized subcutaneously or intramuscularly with affinity purified recombinant Pbs21(1-213), Pbs21(1-188) or native Pbs21 proteins. Following two immunizations, native Pbs21 induces higher titres when administered by either route, than the recombinant protein bearing an insect GPI anchor, which in turn is markedly more immunogenic than the recombinant polypeptide lacking a GPI anchor. When specific anti Pbs21 antibody titres exceeded 1 mg/ml all three antigens were capable of inducing transmission blockade > or = 90%, below 1 mg/ml blockade did not correlate with antibody concentration. PMID- 11012977 TI - Evidence for determining parasitic factors in addition to host genetics and immune status in the outcome of murine Leishmania infantum visceral leishmaniasis. AB - C.B-17 SCID and congenic BALB/C mice were used to examine Leishmania infantum strain pathogenicity independently of host genetic factors. While parasite loads were significantly higher in immunodeficient mice than in immunocompetent mice, the kinetics of infection during a long-term follow-up were similar, suggesting that intrinsic parasitic factors also influence the outcome of L. infantum infection. PMID- 11012976 TI - HLA-A*0201 restricted CD8+ T-lymphocyte responses to malaria: identification of new Plasmodium falciparum epitopes by IFN-gamma ELISPOT. AB - The role of antigen specific CD8+ T-lymphocytes in mediating protection against sporozoite-induced malaria has been well established in murine models. In humans, indirect evidence has accumulated suggesting a similar protective role for antigen-specific CD8+ T-lymphocytes. Nevertheless, the low frequency of circulating specific cells together with the lack of sensitive methods to quantify them has hampered the direct assessment of their function. Using a combination of short-term cell culture and IFN-gamma ELISPOT, we studied CD8+ T lymphocyte responses to a panel of HLA-A*0201 binding peptides. In addition to confirming the response to already described epitopes, we also identified five new CD8+ T-lymphocyte epitopes. These epitopes are presented in pre-erythrocytic stages gene products of Plasmodium falciparum 7G8 strain and correspond to the following protein segments: circumsporozoite (CS) 64-72, 104-113, 299-308 and 403 411; liver stage antigen (LSA-1) repeat region; sporozoite surface protein 2 or thrombospondin related anonymous protein (SSP2/TRAP) 78-88 and 504-513. Four of these peptides are conserved amongst all published sequences of P. falciparum strains. We conclude that the modified IFN-gamma ELISPOT assay is a sensitive technique to monitor antigen-specific CD8+ T-lymphocyte responses in human malaria which may help in the improvement and assessment of the efficacy of malaria subunit vaccines. PMID- 11012979 TI - Proteolytic activity of cysteine protease in excretory-secretory product of Paragonimus westermani newly excysted metacercariae pivotally regulates IL-8 production of human eosinophils. AB - This study investigated the effect of the excretory-secretory product (ESP) of Paragonimus westermani newly excysted metacercariae (PwNEM) on IL-8 production of human mature eosinophils. Treatment of eosinophils with lower concentrations (0.3 and 1 microg/ml) of the ESP significantly (P < 0.01) induced IL-8 production, whereas treatment of cells with higher concentrations (3 and 10 microg/ml) did not. This effect of the ESP was concentration-dependent. Interestingly, the amount of IL-8 production released into the culture supernatants was inversely correlated with the rate of eosinophil survival. When eosinophils were cultured with the same concentrations of the ESP for 24 h, the ESP resulted in eosinophil death in a dose-dependent manner. To investigate whether high proteolytic activity of proteases in the ESP could cause little production of IL-8, 10 microg/ml of ESP was pretreated with heat at 100 degrees C for 10 min or 56 degrees C for 30 min to reduce its proteolytic activity. IL-8 production of eosinophils incubated with heat-treated ESP was markedly increased comparable to that of cells treated with the lowest concentration used in this study. These findings suggest that the protease in the ESP of PwNEM pivotally regulates IL-8 production by controlling of eosinophil survival, depending on the amount of ESP released in vivo. Thus, the cysteine protease in the ESP of PwNEM could provide a novel role to control recruitment of inflammatory cells in larval-infected lesions in human paragonimiasis. PMID- 11012978 TI - The 14-3-3 protein is secreted by the adult worm of Echinococcus granulosus. AB - The 14-3-3 protein, already described in the metacestode of Echinococcus multilocularis, has been characterized in the Echinococcus granulosus adult worm. Immunolocalization studies show the presence of the 14-3-3 protein in the periphery of testes and externally associated with the apical rostellum and adjacent worm tegument. The alcian blue staining in consecutive parasite sections gave similar reactivity patterns, suggesting that the 14-3-3 protein is produced and secreted by rostellar glands. Immunoblot analysis showed the presence of the 14-3-3 protein in somatic and excretory-secretory worm products with higher and smaller apparent molecular masses, respectively, than those detected in E. multilocularis or E. granulosus metacestode tissues. Conversely, the 14-3-3 protein was not detected in metacestode secretory products. Detection of anti-E. granulosus 14-3-3 reactivity in sera of experimentally infected dogs was achieved at early stages of infection. Specific antibody titres decreased during the course of infection. The possible origin and functions of the 14-3-3 protein produced by the adult worm are discussed. PMID- 11012980 TI - Renal carcinogenesis induced by ferric nitrilotriacetate in mice, and protection from it by Brazilian propolis and artepillin C. AB - The protective effect of Brazilian propolis and its extract Artepillin C against ferric nitrilotriacetate (Fe-NTA)-induced renal lipid peroxidation and carcinogenesis was studied in male ddY mice. Fe-NTA-induced renal lipid peroxidation leads to a high incidence of renal cell carcinoma (RCC) in mice. Administration of propolis by gastric intubation 2 h before or Artepillin C at either the same time, 2 h, or 5 h before the intraperitoneal injection of Fe-NTA (7 mg Fe/kg) effectively inhibited renal lipid peroxidation. This was evaluated from the measurement of renal thiobarbituric acid-reactive substances (TBARS) or histochemical findings of 4-hydroxy-2-nonenal (4-HNE)-modified proteins and 8 hydroxy-2'-deoxyguanosine (8-OHdG). Repeated injection of Fe-NTA (10 mg Fe/kg per day, twice a week for a total of 16 times in 8 weeks) caused subacute nephrotoxicity as revealed by necrosis and pleomorphic large nuclear cells in the renal proximal tubules, and gave rise to RCC 12 months later. A protective effect from carcinogenicity was observed in mice given propolis or Artepillin C. Furthermore, the mice given Fe-NTA only developed multiple cysts composed of precancerous lesions with multilayered and proliferating large atypical cells. Mice treated with propolis and Artepillin C also had cysts, but these were dilated and composed of flat cells. These results suggest that propolis and Artepillin C prevent oxidative renal damage and the carcinogenesis induced by Fe NTA in mice. PMID- 11012982 TI - The world health organization classification of malignant lymphomas in japan: incidence of recently recognized entities. Lymphoma Study Group of Japanese Pathologists. AB - New insights into the immunology and genetics of malignant lymphomas have allowed the recognition of new entities and the refinement of previously recognized disease categories. The relative incidence of these subtypes of malignant lymphoma is also known to differ according to geographic location. In order to clarify the current status of malignant lymphomas in Japan and the relative incidences of their subtypes, 3194 patients were classified according to the new World Health Organization (WHO) classification. Among these were 3025 cases (94.71%) of non-Hodgkin's lymphoma (2189 cases (68.53%) of B-cell lymphoma, 796 cases (24.92%) of T-cell lymphoma) and 141 cases (4.41%) of Hodgkin's lymphoma. The incidences of the major subtypes of non-Hodgkin's lymphoma were 33.34% for diffuse large B-cell lymphoma, 8.45% for marginal zone B-cell lymphoma of mucosa associated lymphoid tissue (MALT) type, 8.05% for plasma cell myeloma, 7.45% for adult T-cell leukemia/lymphoma (ATLL), 6.7% for follicular lymphoma, 6.67% for peripheral T-cell lymphoma of unspecified type, 2.79% for mantle cell lymphoma, 2.6% for nasal and nasal-type T/NK cell lymphoma, 2.35% for angioimmunoblastic T cell lymphoma, and 2.35% for precursor B-cell lymphoblastic leukemia/lymphoma, in decreasing order. The other subtypes comprised less than 2%, mainly precursor T cell lymphoblastic lymphoma/leukemia (1.72%), anaplastic large-cell lymphoma of T and null-cell types (1.53%), and B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (1.31%). The incidence of ATLL was influenced by its high percentage (19.20%) in the south-western Japanese island, Kyushu, an endemic area of human T-cell leukemia virus type 1 (HTLV-1), but which appeared to be lower than that in a previous study. The nodular sclerosis and mixed cellularity types of Hodgkin's disease occupied 1.78% and 1.63%, respectively. These data are distinct from those in Western countries and similar in several ways to those in the East, although the relatively high rate of ATLL was attributed to the geographical difference in the etiologic factor, HTLV-1. PMID- 11012981 TI - Microsatellite instability and alteration of E2F-4 gene in adenosquamous and squamous cell carcinomas of the stomach. AB - Microsatellite instability (MSI) due to defective DNA mismatch repair (MMR) is a form of genomic instability underlying the tumorigenesis of various human neoplasms. To evaluate the roles of MSI in the pathogenesis of gastric carcinomas with squamous differentiation, 17 primary stomach cancer patients (15 adenosquamous and two squamous cell carcinomas) were examined for MSI frequency using five microsatellite markers and the criteria for MSI recommended by the National Cancer Institute Workshop. The molecular causes and consequences of MSI in these neoplasms were further researched through the immunohistochemistry of MMR proteins and the mutational analysis of cancer-associated genes targeted by MSI, respectively. Two of the 17 (12%) cases demonstrated MSI at the most examined loci and were classified as having high level MSI (MSI-H). These tumors also exhibited frame-shift mutations at mononucleotide repeats in the target genes, including TGFbetaRII, IGFIIR, BAX, and hMSH6. It is interesting to note that the mutations of the serine (AGC)13 repeats within the E2F-4 gene were found only in the squamous cell carcinoma portions of them, whereas such alterations were not detected in any of the adenocarcinomatous portions. This suggests that E2F-4 might be implicated in the transformation of adenocarcinoma into squamous cell carcinoma and further studies are needed to understand its role in squamous differentiation. PMID- 11012983 TI - Clinicopathological study of pleomorphic xanthoastrocytoma: correlation between histological features and prognosis. AB - The correlation between histopathological characteristics and prognosis was studied in six cases of pleomorphic xanthoastrocytomas (PXA) found in five patients. With regard to the duration from onset to the resection of the tumor, and the postoperative course, three cases had a favorable prognosis, although one case fatally recurred and in another, serial CT showed rapid tumor growth for 3 years. The histological characteristics of the favorable group of PXA comprised remarkable degeneration, low mitotic activity and a low MIB-1 labeling index. In contrast, the characteristics of the latter two cases of PXA rarely showed degeneration, had atypical mitoses, increasing mitotic activity and a higher MIB 1 labeling index, which indicates that the findings of degeneration, atypical mitoses, mitotic activity and MIB-1 labeling index correlate with the biological behavior of PXA. However, with regard to histological appearance and clinical course, PXA are tumors with a wide range of biological behavior. PMID- 11012984 TI - Immunohistochemical inactivation of p14ARF concomitant with MDM2 overexpression inversely correlates with p53 overexpression in oral squamous cell carcinoma. AB - The CDKN2 gene encodes two structurally different proteins: a cyclin-dependent kinase inhibitor called p16, which regulates retinoblastoma protein (pRb) dependent G1 arrest, and a cell cycle inhibitor designated p14ARF, which arrests cell growth in G1-S and also in G2-M. Whereas inactivation of p16 has been described as a frequent event in various cancers, including oral cancer, the current function of p14ARF is still poorly understood. A physical association between p14ARF and MDM2 blocks MDM2-induced p53 degradation, resulting in increased levels of p53, which in turn leads to cell cycle arrest. The present study immunohistochemically examined the expression of p16 and p14ARF together with pRb, MDM2 and p53 status in a series of oral cancers. The results showed that p14ARF was frequently absent in the oral cancers (15/37, 41%) as was p16 immunostaining. Concomitant immunopositivity for p14ARF and MDM2 overexpression was frequently observed in a subset of the cancers, whereas an inverse correlation between p14ARF and MDM2 expression and the diffuse staining of p53 was clearly detected. Moreover, the results showed that in most cases of oral cancer (35/37, 95%) at least one protein was altered, and lymph node metastasis was more frequent in the tumors with alterations in both the p16/pRb and p14ARF/p53 pathway (8/16, 50%) than in the tumors with one or no alteration of these two major pathways. PMID- 11012985 TI - Correlation of tumor markers p53, bcl-2 and cathepsin-D with clinicopathologic features and disease-free survival in laryngeal squamous cell carcinoma. AB - Various recognized prognostic factors in squamous cell carcinoma (SCC) of the larynx influence the therapeutic options offered to an individual patient in order to extend the survival expectancy. Additional prognostic indicators are required in specific patient subgroups. The present study used a standard immunohistochemical technique in order to retrospectively evaluate the accumulation of p53 gene product and the immunoreactivity of bcl-2 protein and cathepsin-D as possible prognostic markers of laryngeal SCC. Formalin-fixed, paraffin-embedded tumor materials were obtained from a series of 64 patients with cancer of the larynx. Immunostaining was evaluated by computerized image analysis. The accumulation of p53 protein was found in 57.8% (37/64) of the patients and was associated with large tumor size. The percentage of p53-positive neoplastic cells increased in high-grade carcinomas, particularly when they simultaneously demonstrated cathepsin-D immunoreaction in stromal cells (P = 0.049); bcl-2 immunoexpression was found to be generally limited. Cathepsin-D immunostaining was observed in tumor parenchymal and stromal cells (31.25% and 37.5% of all cases, respectively); it was found to be useful in defining patient subgroups with differences in relapse-free survival. Among patients with posi tive lymph nodes, those with cathepsin-D immunopositive tumor cells were at higher risk for relapsing (P = 0.0395). Although the classical prognostic factors of laryngeal carcinoma retain their predominance, cathepsin-D immunoreactivity may serve as an additional prognosticator in specific patient subgroups. PMID- 11012986 TI - beta-catenin expression and mutational analysis in renal cell carcinomas. AB - beta-Catenin acts as a downstream transcriptional activator of the Wingless-Wnt signaling pathway. The beta-catenin-Tcf complex transactivates the downstream genes that regulate cell proliferation or inhibit apoptosis. The activation of this pathway through stabilization of beta-catenin is caused either by inactivating mutations of adenomatous polyposis coli (APC) tumor suppressor gene or by activating mutations in beta-catenin exon 3. To determine whether the abnormal expression and activating mutations in exon 3 of the beta-catenin gene are implicated in renal cell carcinogenesis, 52 renal cell carcinomas (RCC) were analyzed by immunohistochemistry, polymerase chain reaction-single-strand conformational polymorphism analysis (PCR-SSCP), and direct DNA sequencing. Immunohistochemically, all cases, as well as normal kidneys, showed membranous and/or cytoplasmic staining patterns without nuclear localization. However, the cytoplasmic accumulations of beta-catenin were observed in five (22.7%) of 22 cases of conventional (clear cell) renal carcinoma, but not in papillary or chromophobe renal carcinomas. The beta-catenin mutation was identified in only one case of conventional renal carcinoma and was a single-base missense mutation on codon 61, leading to substitution of glutamine by arginine. In conclusion, this study demonstrates that beta-catenin mutations are a relatively rare event in RCC and that cytoplasmic accumulations of beta-catenin protein are found only in conventional (clear cell) renal carcinomas. These data suggest that the activation of the beta-catenin signaling pathway may partly play a role in the development of conventional RCC. PMID- 11012987 TI - Angiomatoid (malignant) fibrous histiocytoma: a peculiar low-grade tumor showing immunophenotypic heterogeneity and ultrastructural variations. AB - To clarify the cellular differentiation features and facilitate diagnosis of angiomatoid (malignant) fibrous histiocytoma (AFH), four cases of AFH were examined by clinicopathologic, immunohistochemical and ultrastructural analyses. The age of the patients ranged from 10 to 24 years (mean, 17 years) and the sex distribution was equal. All cases were of subcutaneous origin: three arose in the trunk and one in the upper extremity. All patients presented with systemic symptoms, including inflammatory signs and anemia. After a mean follow up of 11 years 3 months, all patients were alive and well, although one patient twice developed local recurrence after surgery. All cases presented as multinodular, cystic and hemorrhagic tumors ranging in size from 4 to 11 cm (mean, 8 cm) and were characterized by sheets of bland spindle or round cells with oval nuclei within a circumscribed nodule often surrounded by a lymphocytic cuff. One tumor showed predominantly round cell morphology similar to Ewing's sarcoma/primitive neuroectodermal tumor. All cases (100%) exhibited immunoreactivity for vimentin, desmin, CD68 and CD57 (Leu-7). Three cases (75%) were positive for synaptophysin, and reactivity for alpha-smooth muscle actin, epithelial membrane antigen, neuron specific enolase and CD99 (O-13) was present in two cases (50%) each. The three cases examined by electron microscopy had a mixture of fibrohistiocytic, myofibroblastic and undifferentiated cells containing cytoplasmic processes and dense-core granules. It is important for accurate diagnosis of this peculiar soft tissue tumor to recognize that it has a variety of immunophenotypes, such as histiocytic, myofibroblastic, epithelial and neural, and may occasionally have a predominantly round cell morphology. PMID- 11012988 TI - Pigmented villonodular synovitis secondary to laceration of the perforating branch of the peroneal artery. AB - A case of peroneal artery injury subsequently developed into a lesion resembling an extra-articular tenosynovial giant cell tumor, which is a type of pigmented villonodular synovitis (PVNS). This case supports the hypothesis that accident trauma, such as a vascular injury, can be the etiology of PVNS. PMID- 11012989 TI - Desmoplastic cerebral astrocytoma of infancy intermingling with atypical glial cells. AB - Despite the rarity of desmoplastic cerebral astrocytoma of infancy (DCAI), it has distinct clinical and pathological features. The present case is a typical DCAI except for its detection and operational age and intermingling with pleomorphic glial cells. In this case, although a cystic lesion of the right temporal lobe was noticed when the patient was 6 months old, it was not regarded as a tumor and wasn't removed until he was 9 years old. It is quite unusual that a DCAI was able to exist in the cerebrum for 9 years. However, no metastasis occurred and distinct macroscopic and microscopic features of the tumor were not different from typical DCAI except for an intermingling with pleomorphic glial cells. Furthermore, even in the pleomorphic areas, the absence of necrosis and an MIB-1 index of 2.9% indicated non-aggressive growth. These features of the present case may provide additional information as to the character of DCAI, which generally has a favorable prognosis. PMID- 11012990 TI - Lymphoid hyperplasia of the large intestine: a case report with immunohistochemical and gene analysis. AB - A case of lymphoid hyperplasia arising in the large intestine of a 54-year-old woman is described. Barium enema X-ray and colonoscopic examination revealed multiple small polyps in the right side of the colon. Pathological findings from forceps biopsy revealed similar features to a mucosa-associated lymphoid tissue (MALT) lymphoma. A right hemicolectomy with mesenteric lymph node dissection was carried out. Histological sectioning demonstrated hypertrophic lymphoid follicles with well-formed germinal centers. Occasionally, lymphocytes infiltrated the crypts, in a way similar to that found in lymphoepithelial lesions, which was suggestive of a MALT lymphoma diagnosis. Cryptitis was also observed in the lamina propria. Immunohistochemically, proliferating lymphocytes were positive for CD20 (L26) and negative for CD45RO (UCHL-1). Analyses of immunoglobulin gene (IgHJH) rearrangement could not detect any monoclonality in these cells. These findings suggested that the present case should be categorized as lymphoid hyperplasia rather than lymphoma. PMID- 11012991 TI - Hepatoid variant of yolk sac tumor of the testis. AB - A case of testicular yolk sac tumor (endodermal sinus tumor) consisting predominantly of hepatoid cells is documented. A mass measuring approximately 4 x 3 cm was noted in the left testis of a 64-year-old man. Preoperative examination revealed an elevated serum level of alpha-fetoprotein (5479 ng/mL). Histologically, the lesion was composed predominantly of sheet-like or trabecular proliferation of hepatocyte-like cells with eosinophilic or clear cytoplasm. The tumor cells were immunoreactive for alpha-fetoprotein, antimitochondrial antibody, cytokeratin (AE1/AE3), alpha-1-antichymotrypsin, alpha-1-antitrypsin, albumin, carcinoembryonic antigen and epithelial membrane antigen. It was necessary to distinguish this variant lesion from metastatic hepatocellular carcinoma, embryonal carcinoma and hepatoid carcinoma. PMID- 11012993 TI - A new era? Back to the future, again. PMID- 11012992 TI - Mesenchymal dysplasia of the placenta. AB - A severe case of placental mesenchymal dysplasia occurred in association with intrauterine fetal death (IUFD). The gravida-1, para-1 mother was a 26-year-old Japanese. The first pregnancy was unremarkable and a healthy female infant was delivered. The present pregnancy had been uneventful until 34 weeks of gestation when IUFD was detected. The 1516-g (mean +/- SD, 2050 +/- 387 g) stillborn infant had no external abnormalities and the karyotype was 46,XX. The placenta was markedly enlarged (1050 g; mean +/- SD, 452 +/- 202 g), and approximately 80% was occupied by extraordinary enlarged villous structures with a myxoid appearance. Histologically, the dysplastic villi had myxoid stroma and a decreased number of, occasionally obliterated, fetal vessels. There was no abnormal trophoblastic proliferation. Large-sized fetal vessels in the chorionic plate frequently contained organized thrombi. This is the first case of placental mesenchymal dysplasia, which possibly lead to the IUFD. PMID- 11012994 TI - Implementation of the Zuluaga-Raysmith (Z-R) model for assessment of perceived basic human needs in home health clients and caregivers. AB - The Zuluaga-Raysmith (Z-R) model is a conceptual framework that incorporates accepted concepts of universal basic human needs developed by Maslow, yet removes the hierarchical nature of these. The Z-R model recognizes the existence of a health-illness continuum and accepts that an entity (individual, family, aggregate, or community) may move freely in the direction of greater health and self-actualization or towards illness and premature death. The Z-R model identifies 10 basic needs and recognizes that a perceived deficit in any one of these needs can adversely affect the level of wellness of the entity being considered. This exploratory and descriptive study used 11 nurses as interviewers. Subjects consisted of a convenience sample of homebound clients of a home health agency in a metropolitan city, and selected caregivers (n = 27). A modified functional wellness inventory (developed in 1993 by Louvenia Carter) was used with several open-ended questions, which together related to the 10 needs of the Z-R model. Reliability coefficient of the instrument was 0.84. Descriptive statistics were used to analyze the data, using means, percentages, and frequencies. Open-ended questions were grouped according to content and ranked in order of frequency. The five most pressing needs of this small sample were income; physical health; opportunity to make a contribution; mobility; and mental, emotional, social, and spiritual health (MESSH). Nurses unanimously reported that use of the instrument and the Z-R model helped them to focus on the total person, identify strengths in their clients, identify perceived needs deficits, and therefore, with the client, facilitate the preparation of a timely and cost-effective interdisciplinary plan of care to help the entity to move to a higher level of wellness despite the presence of chronic disease, disability, or impending death. These findings suggested that further research is warranted to explore the use of the Z-R model. A replication study is in progress. PMID- 11012995 TI - Cost analyses of home care and nursing home services in the southern Taiwan area. AB - This study compares the cost of long-term care provided at patient homes with that of long-term care provided in nursing homes in southern Taiwan. Caring for a patient with a high degree of dependence at home is more expensive than caring for a patient in a nursing home facility when family costs and provider costs are considered together. This phenomenon is not demonstrated for patients with medium degrees of dependence. To be cost-effective, home care services should target patients with medium physical disability, and nursing home care should focus on patients with high levels of dependence. PMID- 11012996 TI - The crisis nature of health care transitions for rural older adults. AB - The complex health, socioeconomic, and environmental problems experienced by many American elders often place them at high risk for disease and disability. Over time, acutely or chronically ill older persons experience numerous transitions across various health care settings. Although availability of health services is improving in rural areas, barriers such as distance, geography, and poor distribution often limit access to health care. In a longitudinal rural ethnography, the health care transition experiences of older adults, families, and health care providers were examined. A major ethnographic theme emerged from analysis data from interviews, participant observations, and photographs: the crisis nature of health care transitions experienced by rural older adults and their families and observed by rural nurses and other health care providers. Several patterns were observed including the crisis was compounded by surprise; limited knowledge of local resources exacerbated the crisis; inconsistent discharge planning disrupted transitions; changing family support necessitated admission to nursing homes; continuity of care in nursing home discharge lessened transition crisis; and rural home health care was identified as a strength. Recommendations were made for community-based interventions to improve the transition experience. Comprehensive care management services provided by public health nurses (PHNs) in the local rural community were recommended. PMID- 11012997 TI - Extending the extended family for homeless and marginally housed African American women. AB - Young homeless African American women and elderly marginally housed African American women have health, housing, and personal concerns specific to their age cohort, yet they also have parallel and complementary needs. The young struggle to find affordable housing, while the old may have difficulty in maintaining their homes. This article reports select findings from a pilot study designed to describe these two groups of women. The preliminary study was conducted preparatory to the development of a larger study to explore factors that would facilitate or hinder linking the two groups of women for mutual assistance in housesharing arrangements. Interviews and housing history findings revealed contrasts and similarities among the women and between both cohorts that reflected individual differences, common yet divergent life courses, and collective responses to family life situations, societal trends, and policies. Advantages and disadvantages of housesharing were delineated with 56.3% of the homeless women and 81.3% of the elderly women viewing coresidential living as an option worth considering. Housesharing arrangements should be further investigated by nurses and colleagues. Findings from this study are foundational for establishing alliances that may be a means to promote health and strengthen "family" in both populations. PMID- 11012998 TI - Missouri rural adolescent pregnancy project (MORAPP). AB - The purpose of this study was to investigate the pregnancy outcomes and services available to adolescent women in Missouri, comparing rural and urban residents. A secondary analysis of a large public use data set obtained from the Missouri Department of Health was done for the 5-year period 1992 to 1996. Data were collected by county, with each of the 114 counties of the state classified as rural or urban. The SAS program was used for analysis. Results demonstrated definite patterns of similarity and dissimilarity among the teen mothers based on residence, when age and race were controlled. There were over 54,000 births to adolescent mothers in the 5-year study period. Among the outcomes of pregnancy studied were: abortion rates, inadequate weight gain, intrauterine growth retardation, and low-birthweight (LBW) infants. Among the services available to the young mothers that were studied were fertility services, WIC (women, infants, and children supplemental nutrition) program, food stamps, and Medicaid coverage for pregnancy and infant care. The outcomes are probably generalizable to Midwestern, rural/urban states similar to Missouri. PMID- 11012999 TI - A substance use prevention framework: considering the social context for African American girls. AB - Shifting patterns of substance use (that is, early initiation, increased marijuana use, narrowing differences in gender use) and the disproportionate socioeconomic obstacles that are related to substance use among ethnically diverse adolescent females create the need to develop ethnic and gender-specific substance use prevention frameworks. This article describes and applies a substance use prevention framework to African American females. Gender socialization and self-efficacy are presented as key concepts, along with the assertion that every substance use prevention framework should examine the influences of specific societal factors (such as racism, sexism, classism, and ageism) on substance use. Rationale and guidelines for designing ethnically sensitive and gender-specific research projects and intervention programs regarding substance use prevention are offered. Public health nurses (PHNs) are uniquely positioned to use this framework in their work with African American adolescent girls, specifically, and in general with other ethnically diverse groups. PMID- 11013001 TI - The clinical specialist in community health nursing: a solution for the 21st century. AB - The clinical specialist (CS) in community health nursing (CHN) addresses health problems in populations and communities in ways that are different from and complimentary to strategies used by practitioners of individualized patient care. Four programs based on comprehensive assessments, systematic program planning, deliberate implementation, and both formative and summative evaluation are presented. The programs were developed by graduate students in the CHN clinical specialization major at the Frances Payne Bolton School of Nursing. With expert faculty guidance and a competency-driven program of study, strategies were implemented to promote the health of community-dwelling well elderly people, home health care nurses who provide service to patients with respiratory problems, and high school students in a private religious school. Experienced parish nurses also benefited from a program designed to enhance their skills in theological reflection. This article also describes the curricular design developed to educate this clinician for the 21st century. Lastly, continued attention to the dynamic criteria of the American Nurses Association Standards of Community Health Nursing Practice is recommended as crucial to the further development of this clinical specialization. PMID- 11013000 TI - Community resiliency: the potential for community health nursing theory development. AB - There is insufficient theory development within community health nursing that addresses the community as a collective and not simply as groupings of aggregates. Conceptual clarification of nursing's domains is needed, however, before theory development can take place. In an attempt to rectify this situation, this article discusses the environment domain. Community resiliency, or the ability of a community to deal with adversity and in so doing reach a higher level of functioning, is presented as a useful framework to gain greater understanding of this particular domain. Findings from research that have resulted in a community resiliency model for describing the environment domain, within which rural-based community health nurses (CHNs) work, are presented. The relevance for community health nursing theory is included with recommendations for future research and subsequent theory development. PMID- 11013002 TI - Health protecting and health promoting behaviors of African Americans living in Appalachia. AB - The importance of the influence of health protecting and health promoting behaviors on the health of Appalachian African Americans has not been widely published. Review of the literature revealed a paucity of research data describing these behaviors. Therefore, the purpose of this descriptive study was to describe health protecting and health promoting behaviors of Appalachian African Americans. Pender's (1996) health promotion conceptual framework was used as the organizing framework for this study. Data were collected through structured interviews with 204 Appalachian African Americans representing two states and six counties in Appalachia. Quantitative research methodology was used to describe health protecting and health promoting behaviors. Data analysis revealed that these participants actively engaged in health protecting and health promoting behaviors. Morbidity data, mortality data, and family history of diseases were consistent with those reported nationally. Findings from this study support the need for a long-range research study of Appalachian African Americans. PMID- 11013003 TI - Osteoporosis education programs: changing knowledge and behaviors. AB - Osteoporosis is an age related metabolic disease that primarily affects women and causes bone demineralization that results in fractures. Early identification of risk factors for osteoporosis and development of prevention programs is needed to halt the increasing incidence of the disease. Public health nurses (PHNs), with their emphasis on primary, secondary, and tertiary prevention with individuals and families, are in a unique position to protect the health of these vulnerable populations who are at risk for osteoporosis. This article describes the implementation and program evaluation of three osteoporosis prevention educational programs that use three levels of intensity of design. Each design is based upon the learning needs of the targeted audience. The goals of each program were to increase knowledge of osteoporosis, increase health beliefs, and increase the frequency of osteoporosis preventing behaviors. Theoretical aspects from adult learning and the Health Belief Model (HBM) were used to develop the programs. For the program evaluation, participants completed evaluation instruments before and 3 weeks after participating in an osteoporosis health education program. Participants in all programs had significantly higher levels of knowledge after completing the programs; however, overall, there was no change in health beliefs or behaviors. Implications of these findings are discussed. PMID- 11013004 TI - Molecular analysis of human immunoglobulin heavy chain variable region associated determinants recognized by anti-VH3 antibodies 7B4, B6 and D12. AB - 7B4, B6 and D12 are murine monoclonal antibodies (MoAb) that bind to some human immunoglobulin heavy chain products of the closely related V3-30, V3-30.3 and V3 33 genes from the VH3 family. B6 and D12 have additional reactivities with some immunoglobulins (Ig) encoded by the V3-11 and V3-7 genes; D12 also reacts with some V3-43 gene Ig. We show here, by site-directed mutagensis, that the lysine at position 57 in the complementarity-determining region 2 (CDR-2) of the V3-30 gene product is crucial for epitope recognition by all three anti-VH3 MoAbs. Further analysis of the amino-acid sequences of a large panel of Ig reactive, or nonreactive, with MoAb 7B4 indicates that the determinant recognized by 7B4 is dependent on the presence of the tetrapeptide sequence NKYY between positions 56 and 59 in the CDR-2. Comparing the efficiency of 7B4 reactivity with VH3 gene encoded human Ig indicates that amino-acid position 4 in the frame region 1 (FR 1) may also influence the binding of 7B4 to Ig encoded by three very closely related germline genes, V3-30, V3-30.3 and V3-33. NKYY is also found on the gp120 V3 region of human immunodeficiency virus (HIV)-2, SIV and HTLV-4. We also report that other tetrapeptide sequences found on the 56-59 motif of heavy chain variable regions encoded by germline genes are expressed on the solvent exposed V2 region of gp120 of HIV-1 isolates. The possible significance of these observations is discussed. PMID- 11013005 TI - Protective effect of Plantago major L. Pectin polysaccharide against systemic Streptococcus pneumoniae infection in mice. AB - The antibacterial effect of a soluble pectin polysaccharide, PMII, isolated from the leaves of Plantago major, was examined in inbred NIH/OlaHsd and Fox Chase SCID mice experimentally infected with Streptococcus pneumoniae serotype 6B. Serotype 6B is known to give a more protracted infection when injected intraperitoneally into susceptible mice than more virulent serotypes like type 4. PMII was administered i.p. either once 3 days before challenge or once to thrice from 3 to 48 h after challenge. The number of bacteria in blood and the mouse survival rate were recorded. Pre-challenge administration of PMII and also lipopolysaccharide (LPS), included as a control, gave a dose-dependent protective effect against S. pneumoniae type 6B infection. However, injection of PMII after establishment of the infection in NIH/OlaHsd mice had no effect. The data demonstrate that, firstly, the polysaccharide fraction PMII from P. major protects against pneumococcal infection in mice when administered systemically prechallenge, and secondly that the protective effect is owing to stimulation of the innate and not the adaptive immune system. PMID- 11013006 TI - Splenectomy and adoptive cell transfer reveal a prominent role for splenic memory lymphocytes in the development of chronic relapsing experimental autoimmune encephalomyelitis. AB - We previously reported that acute experimental autoimmune encephalomyelitis (EAE), induced by active immunization of SJL mice, could be converted into chronic relapsing EAE (CR-EAE) by a pretreatment with neuroantigen and killed mycobacteria 2 months earlier. This finding indicates that immune memory, established by the pretreatment, influences the subsequent EAE induction. The present study shows that splenectomy and lymphadenectomy, applied 1 week before the subsequent active immunization of the pretreated mice, efficiently abort the chronic nature of CR-EAE. Furthermore, we have found that adoptive transfer of lymphocytes from the spleen (but not of those from the local draining lymph nodes) of the pretreated mice to naive syngeneic recipients 1 week before the acute EAE-induction immunization results in the development of CR-EAE. On the other hand, the transfer of lymphocytes from the local draining lymph nodes aggravates the acute disease. These data support a critical role for immune memory of the previous suboptimal challenge in the development of chronic relapsing demyelinating disease. PMID- 11013007 TI - Neutrophil response of transgenic mice expressing human group IIA phospholipase A2 in bacterial infections. AB - Group IIA phospholipase A2 (PLA2) is a newly recognized acute phase protein with marked antibacterial properties. We have shown previously that transgenic C57BL/6 J mice expressing human group IIA PLA2 (PLA2+ mice) are more resistant to bacterial infections than nontransgenic C57BL/6 J mice that, among mice, are unusual in that they lack the mouse analogue of group IIA PLA2 (PLA2- mice). To elucidate the possible mechanisms involved in the host response of these mice in bacterial infection, peripheral inflammatory cell responses of PLA2+ and PLA2- mice were studied after i.p. administration of Escherichia coli, E. coli lipopolysaccharide or Staphylococcus aureus. Uninfected PLA2+ mice had higher numbers of lymphocytes and polymorphonuclear neutrophil leukocytes (PMNs) in their blood than PLA2- mice. In PLA2+ mice, the number of PMNs increased in peripheral blood in parallel with the concentration of group IIA PLA2 after the administration of bacteria, whereas these responses were not seen in PLA2- mice. High concentrations of group IIA PLA2 in PLA2+ mice may increase the synthesis of bioactive molecules, such as prostaglandins, which in turn may mobilize PMNs into circulation. Our results support the hypothesis that group IIA PLA2 is an important inflammatory mediator in bacterial infections. PMID- 11013008 TI - Hypersusceptibility of A/J mice to tuberculosis is in part due to a deficiency of the fifth complement component (C5). AB - Mycobacterium tuberculosis (MTB) causes tuberculosis in man, which occurs as an acute, chronic or dormant disease reactivating over several years. The mechanisms of persistence and reactivation are not well understood and there is a need for animal models. Moderate-dose, aerosol infection killed A/J mice earlier than partially resistant C57Bl/6 mice, whereas a low-dose, aerosol-induced chronic infection exacerbated earlier in A/J mice. A/J mice lethally infected with MTB but drug cured of disease underwent reactivation of tuberculosis at least 100 days before similarly infected C57Bl/6 mice. Because A/J mice were C5 deficient, congenic B10 mice sufficient and deficient for C5 were infected intravenously with MTB to define the role of C5. C5-deficient mice again showed enhanced growth of MTB in the lungs. MTB-infected macrophages from C5-deficient mice showed enhanced growth of MTB coinciding with a reduced secretion of both cytokines (TNF alpha, IL-1beta, IL-6, IL-12) and chemokines (KC, MIP-2 and MIP-1alpha) in A/J and TNF-alpha and chemokines in C5-deficient mice. Because C5-deficient macrophages could be activated from extraneous C5 and TNF-alpha we suggest that both play a role in the macrophage-mediated killing as well as containment mechanisms in tuberculosis. PMID- 11013009 TI - The Vdelta1 T cell receptor repertoire in human nasal mucosa. AB - The gamma/delta T cells play important roles as the first line of defence in mucosal immunity. Vdelta1 bearing T cells comprise the major population of gamma/delta T cells in the human nasal mucosa. Recently, it was reported that certain receptors are predominant at given anatomical locations. The purpose of this study was to elucidate the gamma/delta T-cell receptor repertoire in human nasal mucosa. Nasal epithelia were scraped and peripheral blood were obtained from three healthy volunteers. We performed reverse-transcription polymerase chain reaction (RT-PCR) analysis and characterized the junctional sequences of Vdelta1 T-cell receptor (TCR) transcripts. Vgamma and Vdelta gene usage in intraepithelial lymphocytes (IELs) in right and left nasal mucosa were almost the same, but they were different from those in peripheral blood lymphocytes (PBLs). The VDJ sequence of nasal Vdelta1 T-cell receptor transcripts showed diversity and were different for each individual. Vdelta1 in the IELs were rearranged to Jdelta1 and Vdelta1 in the PBLs were rearranged not only to Jdelta1 but also to Jdelta3. Vdelta1 sequences in the IELs in the nasal mucosa showed no overlap with those in the PBLs. These results indicate that gamma/delta T cells in the nose are organ specific lymphocytes. PMID- 11013010 TI - Down-regulation of interleukin-12, interleukin-12R expression/activity mediates the switch from Th1 to Th2 granuloma response during murine Schistosomiasis mansoni. AB - In murine schistosomiasis mansoni the worm egg-induced granulomatous inflammation is bi-phasic: an initial Th1 type is subsequently switched to a Th2 type response. Analysis of the cellular, molecular base of the Th1-associated response (5-6 weeks post infection) revealed mRNA messages for interleukin (IL)-12 p40, IL 12Rbeta2 and interferon (IFN)-gamma in the granulomatous livers. When the Th2 type granulomas matured (8 weeks post infection) message expression weakened or became extinct. Macrophages of the Th1 type granulomas produced maximal amounts of IL-12, but production diminished in the mature granulomas. A similar pattern of IL-12 responsiveness of granuloma lymphocytes was observed. In vitro IL-12 production by Th1 type granuloma macrophages was enhanced by tumour necrosis factor (TNF)-alpha and IFNgamma, whereas lymphocyte IL-12 responsiveness was boosted only by TNF-alpha. Both systems were down-regulated by IL-4 and IL-10 cytokines. Treatment of mice with anti-IL-10 monoclonal antibodies (MoAb) between 6 and 7 weeks of the infection enhanced mRNA expression for IFN-gamma and IL 12Rbeta2, but not for IL-12 p40. It is concluded that IL-12 and IL-12R expression and function regulate the Th1 phase of the liver granulomatous response. This phase is cross-regulated by type-2 cytokines especially IL-10. PMID- 11013011 TI - Tamoxifen decreases renal inflammation and alleviates disease severity in autoimmune NZB/W F1 mice. AB - It has been documented that sex hormone may play a role in the pathogenesis of murine lupus. To determine the effect of tamoxifen (TAM) on NZB/W F1 female mice, a total dose of 800 microg (22 mg/kg body weight) of TAM was administered subcutaneously every 2 weeks. The control mice were injected with peanut oil only. After treatment with TAM for 5 months, the mice were killed and immunological parameters were evaluated. The results suggest that NZB/W F1 mice treated with TAM had less severe proteinuria and increased survival rate compared to controls. Flow cytometric analysis of splenocytes revealed a significantly lower percentage of B cells and CD5+ B cells in the TAM-treated group. There was a significantly lower serum level of soluble tumour necrosis factor (TNF) receptor I and II molecules in the TAM-treated mice. Immunohistological study showed that control mice had severe immune complex deposition in the kidney. In contrast, TAM-treated mice had much less pathological change. In summary, this study demonstrated that TAM treatment might be able to alleviate the symptoms of lupus nephritis, influence B-cell count, modulate the expression of cytokine receptors and thereby subsequently affect immune function. Further studies to determine the cellular mechanisms in lupus nephritis may increase our understanding of this complex disease and provide additional targets for therapeutic intervention. PMID- 11013012 TI - Analysis of the tissue distribution of the rat C5a receptor and inhibition of C5a mediated effects through the use of two MoAbs. AB - The C5-anaphylatoxin C5a is a protein of 74 (human) or 77 (rat) amino-acid residues, respectively, the generation of which may be induced by either the classical and/or the alternative pathways. C5a binds specifically to its receptor (C5aR/CD88) which belongs to the superfamily of G-protein-coupled receptors with seven transmembrane segments. In this study we describe the tissue distribution of the rat C5aR (rC5aR) and the blocking of its ligand by the application of two monoclonal antibodies (MoAbs). The first antibody (MoAb R63) which is directed against the amino-terminal domain Ex1 of the rat C5aR was generated in mice immunized with RBL-2H3 cells which had been stably transfected with the rat C5a receptor gene. Checking the rC5aR expression in various tissues bronchial epithelial cells stained positive only in tissue samples from animals with a mycoplasm infection indicating that the receptor may be induced in this cell type as a consequence of an inflammatory process. Using immunohistochemistry there was no evidence for nonmyeloid expression in the large or small intestine, heart, lung, kidney or liver of the normal rat. The MoAb R63 was found to be a reliable tool for the investigation of the expression of the receptor by FACS analyses or immunohistochemistry. Despite numerous attempts neutralizing antibodies could not be generated against the receptor. Therefore a C5a-ligand neutralizing MoAb was generated against the synthesized carboxyterminal 20mer peptide. This antibody (6 9F) recognized the carboxy terminus of C5a/C5a-FLUOS and prevented its binding at a three-fold molar excess as evidenced by FACS-analyses. It also blocked the C5a mediated signal transduction as demonstrated by the inhibition of intracellular Ca2+-release (at a 16-fold molar excess) and the release of N-Acetyl-beta-D glucosaminidase (at a 25-fold molar excess). PMID- 11013013 TI - Detection of a soluble form of the complement membrane attack complex inhibitor CD59 in plasma after acute myocardial infarction. AB - Activation of the complement system has been documented in both experimental and clinical studies of acute myocardial infarction (AMI). Our earlier immunohistochemical studies have shown that the deposition of the membrane attack complex (MAC) of complement is associated with the loss of protectin (CD59), a glycosyl-phosphatidylinositol (GPI)-anchored sarcolemmal regulator of MAC, from the human and rat infarcted myocardium. In this study we detected, using an enzyme immunoassay (EIA), CD59 in the plasma of AMI patients at a concentration of 23.0+/-8.4 ng/ml (mean +/- SD; n = 17) at 4 h and 27.3+/-11.8 ng/ml (n = 24) at 24 h after AMI. Both values were significantly higher than in healthy controls (7.8+/-6.4 ng/ml; n = 20; P<0.001). The amount of CD59 correlated with the level of soluble terminal complement complexes (SC5b-9; r = 0.84; P<0.01) in the plasmas of AMI patients. Our results suggest that myocardial damage leads to release of CD59 from the sarcolemmal cell membranes during AMI. PMID- 11013014 TI - The function of a journal. PMID- 11013015 TI - An economic assessment of Apligraf (Graftskin) for the treatment of hard-to-heal venous leg ulcers. AB - Several recent advances in wound care may offer promise for the treatment of hard to-heal venous leg ulcers. One such treatment is Apligraf (Graftskin), a bilayered, living human skin construct. To assess the economic impact of Graftskin, a model was constructed to compare the annual medical costs and cost effectiveness of treating hard-to-heal venous leg ulcers with Graftskin vs. compression therapy using Unna's boot. A semi-Markov model was used to describe the pattern of ulcer treatment, healing, and recurrence among patients with venous leg ulcers. Patients received 1 of 2 treatment regimens, Graftskin or Unna's boot, and were followed in the model for a 12-month period. The analysis was done from the perspective of a commercial health plan; therefore, only direct medical costs were included. Health care resource use included the primary therapeutic intervention, additional compression dressings, physician office visits, home health visits, laboratory tests and procedures, management of adverse events, and hospitalizations. The model estimated the annual medical cost of managing patients with hard-to-heal venous leg ulcers to be $20,041 for those treated with Graftskin and $27,493 for those treated with Unna's boot. In addition, treatment with Graftskin led to approximately 3 more months in the healed state per person per year than did treatment with Unna's boot. Because patients treated with Graftskin experienced improved healing compared with those treated with compression therapy using Unna's boot, they required fewer months of treatment for unhealed ulcers. As a result, the use of Graftskin for treating hard-to-heal venous leg ulcers resulted in lower overall treatment costs. PMID- 11013016 TI - Randomized clinical trial of the effect of semi-occlusive dressings on the microflora and clinical outcome of acute facial wounds. AB - This study investigated whether treatment with occlusive, hydroactive polyurethane dressings affects the microflora and clinical outcome of acute wounds. A randomized, controlled trial was performed on 60 patients with acute facial lacerations. Following primary closure, patients were provided with either a dry-gauze dressing or a hydroactive polyurethane dressing (Cutinova Thin). The wounds were assessed clinically and microbiologically prior to closure, then after 5, 28 and 56 days. The dressing, which was removed at day 5, was also sent for microbiological culture. Wounds treated with the polyurethane dressing showed improved comfort and contour (p < 0.04), less erythema (p < 0.03) and less potential for scarring (p < 0.01) at day 5. At day 28 and day 56, there were no significant differences in the clinical assessment between the test and control groups (p > 0.05). A total of 518 isolates were recovered from the patients during the study. Mean number of isolates and bacterial growth density were calculated in both groups. A similar range of microorganisms were obtained from both treatment groups with no clear difference in organism colonization. This study shows the potential usefulness of facial wounds as a human model for studying acute wound healing responses and anti-scarring therapy. While short term, clinical benefits of occlusive dressings were evident in the management of acute wounds, these data clearly show that all studies of scarring following acute wounding should extend over at least 3 months post-injury to allow for spontaneous improvement to occur. PMID- 11013017 TI - Simple biochemical markers to assess chronic wounds. AB - We investigated the potential for the biochemical analysis of chronic wound fluid to predict healing using simple and widely available analytes in an out-patient clinic setting. Wound fluid was collected from 12 patients attending a leg ulcer clinic and analyzed for a variety of analytes, including lactate, total protein, and albumin. Twelve weeks after collection the wound was assessed for healing (defined as complete healing or greater than 50% reduction in wound size). The median total protein (44.3 +/- 8.8 g/l) and albumin (25.0 +/- 2.3 g/l) concentrations in exudate collected from four healing wounds were significantly higher (p < 0.05) than in exudate from eight nonhealing wounds (median total protein 29.7 +/- 7.6 g/l, median albumin 17.0 +/- 4.3 g/l). No significant difference was observed for lactate. A second specimen of wound fluid was collected from four of the patients (three nonhealing and one healing). The protein analysis confirmed the pattern observed for the first collection: nonhealing wounds had total protein and albumin which remained low compared to healing wounds. No wound with an exudate albumin of less than 20 g/l healed. Both total protein and albumin are stable analytes which can be easily measured in any laboratory and may offer a simple biomarker of healing in chronic wounds. PMID- 11013018 TI - In vitro fibroblast populated collagen lattices are not good models of in vivo clinical wound healing. AB - In chronic wounds, the healing process is prolonged and incomplete, proceeding in an uncoordinated manner, and resulting in poor anatomical and functional outcome. There have been numerous attempts to discover models that mimic human wound healing processes. The fibroblast populated collagen lattice is one such model that has been proposed. This study evaluated whether the fibroblast populated collagen lattice can be a model of chronic wound healing using the pressure ulcer as a paradigm. Fibroblast cultures of wound biopsies and wound volume measurements were obtained serially during a four arm blinded, placebo-controlled sequential cytokine clinical trial of pressure ulcers. Fibroblasts obtained from study patients were added to collagen lattices and contraction was determined daily for 10 days. Collagen gel-area measurements were converted to reflect percentage of gel contraction. These data of both edge and base wound biopsies on days 0, 10, and 36 were categorized into treatment groups and one-way analysis of variance showed no significant differences in contraction among these groups. When considering all fibroblast populated collagen lattices, there was significantly greater contraction at days 10 and 36 for cells from both edge and base biopsies compared to day 0 (p < 0.05). The Spearman Rank Correlation test comparing all patients with fibroblast populated collagen lattice results from fibroblasts obtained at the edge or base of the wound at days 0, 10, and 36 and clinical pressure ulcer healing on day 36 showed no correlation. This lack of correlation not only persisted for each of the four treatment arms but also for responder status based on decrease in wound volume over the 35 day trial period. In conclusion, chronic wound healing is a complex process that is not modeled by in vitro fibroblast populated collagen lattices. PMID- 11013019 TI - Denervation retards but does not prevent toetip regeneration. AB - Toetips of mammals regrow after amputation by a process similar, but not identical, to that which occurs during regeneration of a newt limb. Nerve is needed as a mitotic stimulant for newt limb regeneration but the requirement for nerve during rodent digit-tip regeneration is not known. Nerve dependence in rats was tested by severing the sciatic nerve in one hindlimb, amputating digit-tips from the central digits of both hind feet, and comparing the amount of regrowth in innervated and denervated digits. Denervation delayed soft-tissue wound healing. However, denervation did not significantly affect bone regrowth when animals were examined at one month. Because we suspected delayed bone regrowth, we used a new method that we developed to follow bone growth at several time points in each animal. Termed visible bone fluorescence through nail, this technique used serial injections of fluorescent calcium-deposition markers and observation through the toenails to observe bone growth in living animals. Using this method it was possible to detect retarded bone regrowth in denervated digits. Thus, although denervation of rodent tips delayed both soft tissue healing and bone regrowth, it did not prevent ultimate restitution of the amputated part. This suggests that neurotrophic stimulation in the mammalian digit-tip is not identical to that documented during newt limb regeneration, and that growth stimulation may be provided by tissues other than nerve. PMID- 11013020 TI - Matrix metalloproteinase production in regenerating axolotl spinal cord. AB - In urodele amphibian spinal cord regeneration, the ependymal cells lining the central canal remodel the lesion site to favor axonal regrowth. We profiled the production of matrix metalloproteinases by injury-reactive mesenchymal ependymal cells in vivo and in vitro and found that matrix metalloproteinases are involved in this remodeling process in the axolotl (Ambystoma mexicanum). The production of cell-associated matrix metalloproteinases in vivo was shown to be identical to that in our cultured ependymal cell model system. Activated and zymogen forms of matrix metalloproteinases were identified using zymography, chemical inhibitors of matrix metalloproteinases, and cleavage of propeptides by organomercurials. The principal cellular proteinases consisted of matrix metalloproteinase-2 (gelatinase A) and matrix metalloproteinase-1 (type I collagenase), which display characteristic shifts in molecular weight following proenzyme processing by organomercurials. In addition, ependymal cell conditioned medium contained secreted forms of the enzyme undetectable in situ. Matrix metalloproteinase-9 (gelatinase B) as well as matrix metalloproteinase-2 and matrix metalloproteinase 1 were secreted and casein substrate zymography showed the presence of a small amount of a very high molecular weight matrix metalloproteinase-3 (prostromelysin) secreted into the culture medium. Matrix metalloproteinases were still present at 4 weeks post-lesioning when the ependymal cells have just re epithelialized, but decreased near the completion of regeneration (8 weeks post lesioning). Zymography showed no detectable matrix metalloproteinases in unlesioned cord but the presence of tissue inhibitor of metalloproteinase-1 in intact cord was seen by Western blotting. This study shows that matrix metalloproteinases are associated with urodele spinal cord regeneration and validates the use of our ependymal cell tissue culture model system to evaluate ependymal cell behavior during spinal cord regeneration. PMID- 11013021 TI - Increase in wound breaking strength in rats in the presence of positively charged dextran beads correlates with an increase in endogenous transforming growth factor-beta1 and its receptor TGF-betaRI in close proximity to the wound. AB - We have previously shown that positively charged beads (DEAE A25) increase wound breaking strength in linear incisions in rats and nonhuman primates at days 10-14 post-wounding. The increased wound strength may result in part from a stimulation of cells adjacent to the DEAE A25 beads to produce growth factors important for wound healing. In this report, we investigate this hypothesis by comparing the relative expression levels of transforming growth factor-beta1 and its receptor transforming growth factor-beta receptor type I in DEAE A25-treated and contralateral untreated rat linear incisions. DEAE A25-treated incisions were stronger than untreated control wounds at 3 days post-wounding, and the difference in breaking strength reached statistical significance at days 5, 7 and 10. Immunohistochemical analysis revealed a significant increase in transforming growth factor-beta1 and transforming growth factor-beta receptor type I expression in DEAE A25-treated incisions, up to 7 days post-wounding, as compared to untreated control wounds. FACS analysis revealed that macrophage cell lines exposed to DEAE A25 in vitro upregulate transforming growth factor-beta1 and transforming growth factor-beta receptor type I expression by 2-3 fold. Therefore, the increase in expression of transforming growth factor-beta1 and transforming growth factor-beta receptor type I in DEAE A25-treated incisions may be due to an increase in the concentration of macrophages adjacent to DEAE A25 beads, as well as the stimulation of individual macrophages to produce greater amounts of transforming growth factor-beta1 and transforming growth factor-beta receptor type I. This study also supports the significance of transforming growth factor-beta1 in wound healing. PMID- 11013022 TI - Development of a chronic skin defect model and a study of cytokine secretion using the model. AB - In this study, we established a delayed healing chronic type wound model in order to investigate the etiology of chronic wound healing, including wound contraction. Establishment of the model was important for clarification of the mechanism(s) of chronic wound healing and wound contraction and for use in evaluating therapeutic efficacy. A pedicled skin flap was raised beneath the panniculus carnosus membrane on the backs of mice, and after the loose connective tissue at the base of the flap was completely removed surgically, the flap was replaced and sutured. Seven days after surgery, a full-thickness defect measuring 1.5 x 1.5 cm was made in the center of the skin flap. At that time, a defect of the same size, including the panniculus carnosus membrane, was made in another group of mice as controls, and changes with time in wound area were compared between the two groups. The exudate retained on the wound surface was collected, and various cytokines contained in the exudate were measured. In the control group, the wound rapidly contracted and almost completely epithelialized and closed 21 days after surgery. On the other hand, the wound area was significantly larger in the delayed model than in the control animals during the observation period, revealing a delay in wound contraction. Transforming growth factor-beta, interleukin-1beta, and tumor necrosis factor-alpha in the exudates from the wound of the model were significantly higher than in those of the control group, whereas interleukin-6 was low in the model. From these results, it was concluded that this model could be a useful experimental system for studies on wound contraction as well as clarifying the mechanism of so called chronic type wounds. PMID- 11013023 TI - Presidential address: where science touches man, it turns to art. PMID- 11013024 TI - Is routine use of the intensive care unit after elective infrarenal abdominal aortic aneurysm repair necessary? AB - INTRODUCTION: Postoperative care after infrarenal abdominal aortic aneurysm (AAA) repair has traditionally involved admission to the intensive care unit (ICU). With the advent of endovascular AAA repair, the management of open procedures has received increased scrutiny. We recently modified our AAA clinical pathway to include selective use of the ICU. METHODS: Consecutive elective infrarenal AAA repairs performed by members of the vascular surgery division at a university medical center from 1994 to 1999 were analyzed retrospectively with a computerized database, the Medical Archival Retrieval System. Group I consisted of 245 patients who were treated in the ICU for 1 or more days, and Group II included 69 patients admitted directly to the floor. Ruptured, symptomatic, suprarenal, endovascular, and reoperative repairs were excluded. Outcome variables were compared over the 6-year period. RESULTS: Floor admissions increased over the study period with 0%, 0%, 3.3%, 16.3%, 48.6%, and 43.6% of patients admitted directly to the surgery ward from 1994 to 1999. The average ICU length of stay declined from 4.6 to 1.2 days, whereas the hospital length of stay decreased from 12.5 to 6.8 days from 1994 to 1999. The change in ICU use had no effect on death (2.4% in Group I vs 0% in Group II). Major and minor morbidity was comparable. Hospital charges were significantly lower for patients in Group II. CONCLUSION: A policy of selective utilization of the ICU after elective infrarenal AAA repair is safe. It can reduce resource use without a negative impact on the quality of care. PMID- 11013025 TI - Carotid endarterectomy: the financial impact of practice changes. AB - PURPOSE: New techniques in the management of extracranial carotid occlusive disease have focused attention on the outcome and economics of carotid endarterectomy (CEA). Changing practice patterns for CEA must be assessed to allow accurate comparisons. The purpose of this study was to evaluate the effect of practice modifications related to CEA on patient outcome and cost data. METHODS: Data on patients undergoing CEAs at a single institution from fiscal year 1992 to 1998 were prospectively collected and entered into a computerized database. Records were reviewed for patient demographics and outcome with regard to stroke and death. Selected years that corresponded to transitions in perioperative management were audited for complete hospital financial information from. RESULTS: We performed 960 CEAs during the study period, with a combined stroke and death rate of 1.1%. Inflation-adjusted hospital costs per patient in 1998 dollars for the years 1992, 1996, and 1998 were $5494, $4476, and $3350, respectively. In 1998, costs for patients who required arteriography were $1825 greater than those operated on during duplex scan examination alone in 1998. Statistically significant differences occurred in the year-to-year comparisons in the use of arteriography, intensive care unit monitoring, same day admissions, and length of stay. There were no statistically significant differences in the stroke and death rate between years. CONCLUSION: Practice changes related to CEA have resulted in significant savings without detriment in patient outcome. Comparisons between CEA and endovascular techniques will need to be evaluated within this context. Given these advances in perioperative management, it will be difficult to justify carotid stenting on the basis of current economic considerations. PMID- 11013026 TI - Cranial and cervical nerve injuries after repeat carotid endarterectomy. AB - BACKGROUND AND PURPOSE: The incidence of cranial and/or cervical nerve injuries after primary carotid endarterectomy (CEA) ranges from 3% to 48%; however, the clinical outcome of these injuries after repeat CEA has not been thoroughly analyzed in the English-language medical literature. This prospective study analyzes the incidence and outcome of cranial nerve injuries after repeat CEA. PATIENTS AND METHODS: This study includes 89 consecutive patients who had repeat CEAs. Preoperative and postoperative cranial nerve evaluations were performed, including clinical examinations (neurologic) and direct laryngoscopy. Patients with vagal or glossopharyngeal nerve injuries also underwent comprehensive speech evaluations, video stroboscopy, fluoroscopy, and methylene blue testing for aspiration. Patients with postoperative cranial nerve injuries were followed up for a long time to assess their recovery. RESULTS: Twenty-five cranial and/or cervical nerve injuries were identified in 19 patients (21%). They included 8 hypoglossal nerves (9%), 11 vagal nerves or branches (12%) (6 recurrent laryngeal nerves [7%], 3 superior laryngeal nerves [3%], and 2 complex vagal nerves [2%]), 3 marginal mandibular nerves (3%), 2 greater auricular nerves (2%), and 1 glossopharyngeal nerve (1%). Twenty-two (88%) of these injuries were transient with a complete healing time ranging from 2 weeks to 28 months (18 of 22 injuries healed within 12 months). The remaining three injuries (12%) were permanent (1 recurrent laryngeal nerve, 1 glossopharyngeal nerve, and 1 complex vagal nerve injury). The recurrent laryngeal nerve injury had a longer healing time than the other cranial nerve injuries. CONCLUSIONS: Repeat CEA is associated with a high incidence of cranial and/or cervical nerve injuries, most of which are transient. However, some of these have a long healing time, and a few can be permanent with significant disability. PMID- 11013027 TI - Use of shunts with eversion carotid endarterectomy. AB - PURPOSE: The purpose of this study was to examine the utility of carotid shunting in the context of eversion endarterectomy. A comparison of patients who underwent carotid endarterectomy by eversion with and without shunts was performed. METHODS: Over a 5-year period, 2724 eversion carotid endarterectomies were performed. In most of these operations patients were under cervical block anesthesia. A shunt was used in 112 eversion endarterectomies (4.1%). Cervical block anesthesia was used in 103 patients (92.0%), general anesthesia was used in 5 patients (4.5%), and 4 patients (3.6%) were converted from cervical block to general anesthesia intraoperatively. The indications for shunting were neurologic deterioration in 99 patients (88.4%) who were under cervical block anesthesia, procedures performed in neurologically unstable or otherwise compromised patients who were under general anesthesia, and the operator's discretion in the remaining eight patients. RESULTS: There was a combined stroke/death rate of 2.7% in the shunt group. These three cases included one death from myocardial infarction and one delayed death due to intracerebral hemorrhage after discharge. Shunt insertion was unrelated to the negative outcome in these two cases. One perioperative major stroke in the shunt group was identified. Follow-up averaged 12.3 months (range, 1-53 months). CONCLUSION: Carotid shunts can be used effectively in the context of eversion endarterectomy. Shunt insertion is not associated with an increased stroke/death rate in these patients. PMID- 11013028 TI - Prevalence of deep venous reflux in patients with primary superficial vein incompetence. AB - PURPOSE: This prospective study was designed to determine the prevalence of deep reflux and the conditions under which it may occur in patients with primary superficial venous reflux and absence of deep venous thrombosis (DVT). METHODS: We studied 152 limbs in 120 consecutive patients in the standing position who had superficial venous reflux with color flow duplex scanning. Limbs with documented evidence of DVT or post-thrombotic vein wall changes during the examination were studied but not included in the analysis. Limbs were divided into those that had at least reflux in the saphenofemoral, the saphenopopliteal, or the gastropopliteal junction and into those with nonjunctional reflux in the superficial and gastrocnemial veins. Peak velocity and duration of reflux were measured. To examine the recirculation theory, we tested the deep veins by occluding and refluxing saphenous veins 10 cm below the sampling site. RESULTS: Thirteen limbs in 11 patients (9%) were excluded because of previous DVT. Of the remaining 139 limbs, 106 (76%) had junctional reflux. Saphenofemoral junction was involved in 89 limbs (84%), saphenopopliteal junction in 18 (17%), and gastropopliteal junction in 7 (4%). In 33 limbs (24%), reflux was detected in the main trunk or tributaries of the saphenous veins alone with no junctional incompetence. Femoral or popliteal reflux was present in 31 limbs (22%). This reflux was segmental in 27 limbs, and it was limited in the junction in 24 limbs. The mean duration of deep venous reflux was 0.9 seconds, it ranged from 0.6 to 3.7 seconds, and it was significantly shorter than that in the superficial veins (2.6 seconds; P <.0001). In the absence of junctional reflux, the prevalence of deep venous insufficiency (DVI) was significantly lower compared with that in limbs with junctional involvement (2 of 33 vs 29 of 106; P =.038). The mean duration of deep venous reflux in these groups was comparable (0.85 seconds vs 0. 91 seconds; P =.44). Occlusion of the incompetent superficial veins reduced somewhat the duration of the deep venous reflux but did not abolish it (0.88 seconds vs 0.82 seconds; P =.072). The presence of DVI was associated with junctional reflux of high peak velocity and long duration. CONCLUSIONS: The prevalence of DVI in patients with primary superficial venous reflux and without history of DVT is 22%. However, this reflux is segmental, mainly in the common femoral vein, and is of short duration. It is associated with the presence of junctional incompetence that has a high peak velocity and long duration. These findings may explain why surgical correction of superficial reflux abolishes DVI. PMID- 11013029 TI - Prospective screening for postoperative deep venous thrombosis in patients undergoing infrainguinal revascularization. AB - PURPOSE: The incidence of deep venous thrombosis (DVT) in patients undergoing infrainguinal bypass graft procedures has not been well documented, and the need for routine prophylaxis remains controversial. The purpose of this study was to prospectively evaluate the risk of postoperative DVT complicating infrainguinal revascularization. METHODS: Seventy-four patients undergoing infrainguinal bypass graft procedures during a 12-month period were prospectively screened for DVT. Bilateral lower extremity venous duplex scan imaging was performed preoperatively and within 1 week and 6 weeks, postoperatively. Routine DVT prophylaxis was not used, with anticoagulation reserved for specific indications. RESULTS: Of the 74 patients screened, three patients (4.1%) had DVT identified on preoperative venous duplex scan imaging and were excluded from the study. Of the remaining 71 patients enrolled, only two patients (2.8%) had postoperative DVT. Postoperative DVT was ipsilateral to the bypass graft extremity in both patients, with involvement of the peroneal vein in one patient and the femoral vein in the other. Although routine prophylaxis was not used, 18 of these patients (25%) were anticoagulated for other indications, with DVT occurring in one patient (5.6%). Of the remaining 53 patients who did not receive postoperative anticoagulation, only one patient (1.8%) had DVT. CONCLUSIONS: According to this prospective study, the risk of postoperative DVT in patients undergoing infrainguinal revascularization is low. Routine prophylaxis is not recommended, with postoperative anticoagulation reserved for specific indications. PMID- 11013030 TI - Adverse consequences of internal iliac artery occlusion during endovascular repair of abdominal aortic aneurysms. AB - OBJECTIVE: Embolization of the internal iliac artery (IIA) may be performed during endovascular abdominal aortic aneurysm (AAA) repair if aneurysmal disease of the common iliac artery precludes graft placement proximal to the IIA orifice. The IIA may also be unintentionally occluded because of iliac trauma or coverage by the endograft. The purpose of this study was to determine the incidence, etiology, and consequences of IIA occlusion during endoluminal AAA repair. METHODS: Over 2 years, 96 patients have undergone endoluminal AAA repair. The details of the operative procedure, reasons for IIA occlusion, perioperative complications, and clinical follow-up were recorded. RESULTS: The IIA was intentionally occluded in 15 patients (16%) to treat 13 common iliac artery aneurysms, one IIA aneurysm, and one external iliac artery aneurysm. The IIA was unintentionally occluded in 9 patients (9%), resulting from traumatic iliac dissection in 5 patients and coverage of the IIA by the endograft in the remaining 4 patients. Three patients had colon ischemia. One patient with a unilateral IIA occlusion had sigmoid infarction necessitating resection. The other two patients underwent intentional occlusion of one IIA followed by unintentional occlusion of the contralateral IIA because of a traumatic iliac dissection. Both had postoperative abdominal pain and distention; rectosigmoid ischemia was revealed through colonoscopy. Conservative treatment with bowel rest and broad-spectrum antibiotics was successful in both cases. Nondisabling hip and buttock claudication occurred in seven patients (32%) at 1 month but resolved by 6 months in three of these patients. CONCLUSION: Embolization of the IIA for iliac aneurysmal disease and unintentional IIA occlusion due to trauma or graft coverage occurs in a considerable number of patients undergoing endoluminal AAA repair. Most patients with unilateral occlusion do not experience colon ischemia or disabling claudication. Therefore, unilateral embolization of the IIA is well tolerated and allows for the endoluminal treatment of patients with both an AAA and an iliac artery aneurysm, thereby expanding the number of patients who can be managed with an endovascular approach. Although acute, bilateral IIA occlusions should be avoided, significant consequences were not observed in our small series of patients. PMID- 11013031 TI - Safety of coil embolization of the internal iliac artery in endovascular grafting of abdominal aortic aneurysms. AB - PURPOSE: During endovascular grafting of an abdominal aortic aneurysm (AAA), iliac limb extension to the external iliac artery may be indicated when the common iliac artery is ectatic or aneurysmal. Preliminary or concomitant coil embolization of the internal iliac artery (IIA) is thus necessary to prevent potential reflux and endoleak. We sought to determine the safety of hypogastric flow interruption in this setting. METHODS: We retrospectively reviewed 156 patients who underwent stent-graft AAA repair at two institutions between February 1, 1998, and January 31, 1999. Coil embolization of one or both IIAs was undertaken when the diameter of the common iliac artery was more than 20 mm to enable limb endograft extension to the external iliac artery. Bilateral procedures were staged. RESULTS: Thirty-nine (25%) of 156 patients were selected for coil embolization of one (n = 28) or both (n = 11) IIAs. The interventions were performed before (n = 31) or during (n = 8) the stent-graft procedure. Complications included groin hematomas in 3 patients, iliac artery dissection in 1, failure to catheterize the IIA in 2, and transient rise in the serum creatinine level in 3. One patient had erectile dysfunction, and five patients (13%) had buttock claudication after unilateral occlusion. Serious ischemic complications were not observed. CONCLUSION: Coil embolization of one or both IIAs appears to be safe in the setting of endovascular grafting of AAA. Buttock claudication is a relatively significant problem and may limit applicability of this strategy to patients who are unfit for standard open repair. PMID- 11013032 TI - Long-term fate of the aneurysmal sac after endoluminal exclusion of abdominal aortic aneurysms. AB - PURPOSE: Shrinkage of an abdominal aortic aneurysm (AAA) is the hallmark of successful endoluminal treatment. Our goal was to prospectively assess the midterm to long-term shrinkage of the AAA sac after endovascular repair. METHODS: A total of 123 patients with AAA underwent endoluminal treatment with the Ancure device at our institution between February 1996 and February 2000. At least a 1 year follow-up was available for 70 of the 123 patients. AAA sac size, presence of endoleaks, calcifications, and outcome data were collected on these patients at 6, 12, 24, and 36 months after repair and compared with the preoperative AAA size and characteristics. All endoleaks found at the 6-month follow-up visit were treated aggressively with embolotherapy. An AAA sac regression of 0.5 cm or more was considered the minimum measurable decrease. Regression of the sac diameter to 3.5 cm or less was considered a complete collapse of the sac. RESULTS: Successful endoluminal repair was accomplished in 119 of 123 patients. The mortality rate was 0.8% (1/123). There was a steady decrease in AAA sac size from baseline (5.56 +/- 0.1 cm), to 6 months (5.0 +/- 0.14 cm, P =.0006), to 12 months (4.65 +/- 0.13 cm, P =.04), and to 24 months (4.26 +/- 0.16 cm, P =.03). At 24 months, 74% (29/39) had a decrease in sac size of 0.5 cm or more, with 28% (11/39) complete collapse. Patients with initial endoleaks had the same likelihood of regression of sac size (> or = 0.5 cm) when compared with the group of patients with no endoleaks at the 24-month evaluation (64% vs 76%, P =.09). CONCLUSION: Endoluminal AAA repair resulted in a significant reduction in sac size that continues up to 2 years. Significant shrinkage occurs as early as 6 months after placement. The initial presence of endoleaks does not predict the lack of sac regression. PMID- 11013033 TI - Transrenal fixation of aortic stent-grafts for the treatment of infrarenal aortic aneurysmal disease. AB - PURPOSE: We evaluated our early experience with the transrenal fixation of aortic stent-grafts to determine the efficacy of this procedure and its effects on renal artery patency and hemodynamics. METHODS: Twenty-eight patients (22 men) had endoluminally placed modular bifurcated stent-grafts with a bare spring structure at the proximal end crossing the origin of both renal arteries; no patient with infrarenal fixation was included for analysis. The mean age of the patients was 75 +/- 7 years (range, 58-86 years); the mean aneurysm size was 5.8 +/- 0.8 cm (range, 4.7-7.2 cm). Eight patients had preoperative or intraoperative angiographic evidence of renal artery atherosclerotic disease, but only four vessels had luminal narrowing of 50% or greater. No complications were noted during stent-graft placement, and all patients have returned for follow-up visits, ranging from 1 to 12 months (mean follow-up, 6 +/- 4 months). Follow-up evaluations included clinical assessment, duplex ultrasound scan of the renal arteries and kidneys, and computed tomographic angiography. RESULTS: No evidence of lobular or sublobular perfusion defects of the renal parenchyma was detected postoperatively. Two patients exhibited postoperative changes in renal artery hemodynamics-one progressing from a 30% diameter reduction to a greater than 60% diameter stenosis at the 12-month follow-up visit and one with a normal renal artery preoperatively having elevated flow velocities indicative of a greater than 60% stenosis at the 1-month visit. Of 19 patients with normal preoperative renal function, only one has had persistently elevated serum creatinine levels. CONCLUSION: We conclude from this experience that the transrenal placement of open stents is safe and effectively excludes the aneurysm, potentially expanding the availability of this technique to more patients with a short infrarenal aortic neck. Long-term follow-up is essential to determine the overall efficacy of this technique and to identify potential effects on renal artery hemodynamics or kidney function. PMID- 11013034 TI - Does the endovascular repair of aortoiliac aneurysms pose a radiation safety hazard to vascular surgeons? AB - OBJECTIVES: Endovascular aortoiliac aneurysm (EAIA) repair uses substantial fluoroscopic guidance that requires considerable radiation exposure. Doses were determined for a team of three vascular surgeons performing 47 consecutive EAIA repairs over a 1-year period to determine whether this exposure constitutes a radiation hazard. METHODS: Twenty-nine surgeon-made aortounifemoral devices and 18 bifurcated devices were used. Three surgeons wore dosimeters (1) on the waist, under a lead apron; (2) on the waist, outside a lead apron; (3) on the collar; and (4) on the left ring finger. Dosimeters were also placed around the operating table and room to evaluate the patient, other personnel, and ambient doses. Exposures were compared with standards of the International Commission on Radiological Protection (ICRP). RESULTS: Total fluoroscopy time was 30.9 hours (1852 minutes; mean, 39.4 minutes per case). Yearly total effective body doses for all surgeons (under lead) were below the 20 mSv/y occupational exposure limit of the ICRP. Outside lead doses for two surgeons approximated recommended limits. Lead aprons attenuated 85% to 91% of the dose. Ring doses and calculated eye doses were within the ICRP exposure limits. Patient skin doses averaged 360 mSv per case (range, 120-860 mSv). The ambient (> 3 m from the source) operating room dose was 1.06 mSv/y. CONCLUSIONS: Although the total effective body doses under lead fell within established ICRP occupational exposure limits, they are not negligible. Because radiation exposure is cumulative and endovascular procedures are becoming more common, individuals performing these procedures must carefully monitor their exposure. Our results indicate that a team of surgeons can perform 386 hours of fluoroscopy per year or 587 EAIA repairs per year and remain within occupational exposure limits. Individuals who perform these procedures should actively monitor their effective doses and educate personnel in methods for reducing exposure. PMID- 11013035 TI - Aortic fenestration for acute or chronic aortic dissection: an uncommon but effective procedure. AB - BACKGROUND: Aortic fenestration is rarely required for patients with acute or chronic aortic dissection. To better define its role and the indications for its use and to evaluate its success at relieving organ or limb malperfusion, we reviewed our experience with direct fenestration of the aorta. METHODS: A retrospective analysis of all consecutive aortic fenestrations performed between January 1, 1979, and December 31, 1999, was performed. Fourteen patients, 12 men and two women (mean age, 59.6 years; range, 43-81), underwent fenestration of the aorta. All patients were hypertensive and had a history of tobacco use. By Stanford classification, there were three type A and 11 type B patients. In the acute dissection group (n = 7), indications for surgery were malperfusion in six patients (leg ischemia, 4; renal ischemia, 5; bowel ischemia, 3) and intra abdominal bleeding from rupture in two. In the chronic dissection group (n = 7), indications for surgery were abdominal aortic aneurysm in 4 patients (infrarenal, 3; pararenal, 1), thoracoabdominal aneurysm in 1, hypertension from coarctation of the thoracic aorta in 1, and aortic occlusion with disabling claudication in 1. RESULTS: Emergency aortic fenestration was performed in seven patients (surgically for 6 and percutaneously for 1). Fenestration level was infrarenal in four and pararenal in three. Concomitant abdominal aortic graft replacement was performed in four patients, combined with ascending aortic replacement (n = 1) and bilateral aortorenal bypasses (n = 1). In two patients, acute fenestration was performed for organ malperfusion after prior proximal aortic replacement (ascending aorta, 1; descending thoracic aorta, 1). Seven elective aortic fenestrations were performed for chronic dissection (descending thoracic aorta, 2; paravisceral aorta, 2; infrarenal aorta, 2 and pararenal aorta, 1). Concomitant aortic replacement was performed in six patients (abdominal aorta, 5; thoracoabdominal aorta, 1). Fenestration was successful at restoring flow in all 10 patients with malperfusion. Operative mortality for emergency fenestration was 43% (3/7). The three deaths that occurred were of patients with anuria or bowel ischemia, or both. There were no postoperative deaths for elective fenestration. At a mean follow-up of 5.1 years, there were no recurrences of malperfusion and no false aneurysm formations at the fenestration site. CONCLUSION: Fenestration of the aorta can effectively relieve organ or limb ischemia. Bowel ischemia and anuria are indicators of dismal prognosis and emergency fenestration in these patients carries a high mortality. Elective fenestration combined with aortic replacement can be performed safely in chronic dissection. Aortic fenestration is indicated for carefully selected patients with malperfusion and offers durable benefits. PMID- 11013036 TI - Visceral pseudoaneurysms due to pancreatic pseudocysts: rare but lethal complications of pancreatitis. AB - OBJECTIVE: Erosion of pancreatic pseudocysts into adjacent vessels is a rare but highly lethal cause of intra-abdominal hemorrhage. Percutaneous angiographic embolization (PAE) of the bleeding artery has recently been advocated as the preferred therapy. This study was undertaken to survey the outcome after treatment of this complication and to make recommendations for its management. METHODS: An 11-year retrospective analysis was performed of all patients treated at a large tertiary care referral center for visceral artery pseudoaneurysms associated with pancreatic pseudocysts. RESULTS: From 1988 to 1998, 256 patients were admitted for complications of pancreatic pseudocysts. Sixteen patients (11 men and 5 women) were identified in whom a pseudocyst had eroded into a major blood vessel with hemorrhage or development of a false aneurysm. The mean age was 45 years (range, 23-67 years). Active bleeding was present in 13 patients, whereas three had evidence of recent hemorrhage. Ten of 16 patients initially underwent operative therapy, four elective and six emergency, whereas six stable patients were initially treated with PAE. Technical failures of the initial treatment or secondary complications required both therapeutic modalities in six patients, which resulted in 13 total surgical interventions and 10 PAEs. The surgical morbidity rate was 62% (8 of 13), whereas that of PAE was 50% (5 of 10). Three deaths occurred after emergency operations, two of which failed to stop the bleeding, accounting for all of the deaths in the series (3 [19%] of 16). A trend was noted toward increased death with necrotizing pancreatitis (P =.07) and emergency surgery (P =.06). Ranson's criteria were not found to be predictive of death in this series. Surgical drainage procedures were required in seven (44%) of 16 patients for infections (n = 3) or mass effect of the pseudoaneurysm (n = 3). The mean size of pseudoaneurysms that required operative intervention for secondary complications was 13.9 cm, compared with 7.7 cm for all others in the series (P =.046). Long-term follow-up was available in all 13 survivors at a mean of 44 months (range, 1-108 months). CONCLUSIONS: The management of pancreatic pseudocyst-associated pseudoaneurysms remains a challenging problem with high morbidity and death rates. Operation and PAE play complementary management roles. PAE is recommended as the initial therapy for hemodynamically stable patients. Surgery should be reserved for actively bleeding, hemodynamically unstable patients; for failed embolization; and for other secondary complications such as infection or extrinsic compression. PMID- 11013037 TI - Suggested treatment protocol for improving patency of femoral-infrapopliteal cryopreserved saphenous vein allografts. AB - PURPOSE: Cryopreserved saphenous vein allografts are used for femoral infrapopliteal bypass graft purposes when adequate autogenous vein is unavailable. Anticoagulation, immunosuppression therapy, or both have been suggested means for improving allograft patency. Immunosuppression has significant cost and morbidity and has produced variable results. Our successful treatment of luminal surface hypercoagulability associated with certain endovascular procedures prompted the use of an anticoagulation protocol prospectively to improve graft patency and limb salvage for patients receiving femoral-infrapopliteal cryopreserved saphenous vein allografts. METHODS: Between September 1995 and October 1999, 24 patients (15 men and nine women) were enrolled in a prospective clinical trial for salvage of 26 severely ischemic lower limbs with femoral-infrapopliteal cryopreserved saphenous vein allograft bypass grafts. All patients were treated with a protocol (aspirin, low-dose heparin, low molecular weight dextran 40, dipyridamole, and warfarin), and no immunosuppressive agents were used. The cryopreserved saphenous vein allografts were matched to patients by ABO and Rh compatibility. Indications for revascularization were ischemic rest pain (n = 8), nonhealing ulcer (n = 13), or focal gangrene (n = 5), and no usable autogenous vein was available. Follow-up ranged from 2 to 35 months (mean, 19 months). We studied the location and type of outflow anastomosis, specific outflow vessel, morbidity, death, secondary procedures (digital/transmetatarsal amputation), and complications related to the treatment protocol. Life table analyses of primary graft patency and limb salvage were compared with other current reported data. RESULTS: Primary graft patency with Kaplan-Meier life table analysis was 96% at 6 months, 87% at 12 months, and 82% at 18 and 24 months. There were no reoperations for acute graft occlusion. One graft underwent late segmental aneurysmal degeneration and rupture. There were no procedure-related deaths or bleeding complications. During late follow up, anticoagulation was discontinued in three patients (12%) because of gastrointestinal bleeding. Limb salvage was 88% at 6 months and 80% at 12, 18, and 24 months. Patients returned to ambulatory status that was limited only by their other comorbidities. CONCLUSION: Femoral-infrapopliteal bypass graft for limb salvage with a cryopreserved saphenous vein allograft can be an acceptable alternative when autogenous vein is not available. Our treatment protocol substantially improved allograft patency and limb salvage when compared with current published data. PMID- 11013038 TI - Incidence and risk factors of late rupture, conversion, and death after endovascular repair of infrarenal aortic aneurysms: the EUROSTAR experience. European Collaborators on Stent/graft techniques for aortic aneurysm repair. AB - OBJECTIVE: The EUROSTAR (European Collaborators on Stent/graft techniques for aortic aneurysm repair) Registry was established in 1996 to collect data on the outcome of treatment of patients with infrarenal aortic aneurysms with endovascular repair. To date, 88 European centers of vascular surgery have contributed. The purpose of the study was to evaluate the results of this treatment in the medium term (up to 4 years) according to the analysis of "hard" or primary end points of rupture, late conversion, and death. PATIENTS AND METHODS: Patients with aortic aneurysms suitable for endovascular aneurysm repair were notified to the EUROSTAR Data Registry Centre before treatment to eliminate bias due to selective reporting. The following information was collected on all patients: (1) demographic details and the anatomic characteristics of their aneurysms, (2) details of the endovascular device used, (3) complications encountered during the procedure and the immediate outcome, (4) results of contrast enhanced computed tomographic imaging at 3, 6, 12, and 18 months after operation and at yearly intervals thereafter, and (5) all adverse events. Life table analysis was performed to determine the cumulative rates of (1) death from all causes, (2) rupture, and (3) late conversion to open repair. Risk factors for rupture and late conversion were identified through regression analysis. RESULTS: By March 2000, 2464 patients had been registered, and their mean duration of follow-up was 12.19 months (SD, 12.3 months). There were 14 patients with confirmed rupture of their aneurysms. The cumulative rate (risk) of rupture was approximately 1% per year. Emergency surgery was undertaken in 12 (86%) patients, of whom five (41.6%) survived. Two patients who were not treated surgically also died, which resulted in an overall death rate of 64.5% (9/14) of the patients. Significant risk factors for rupture were proximal type I endoleak (P =.001), midgraft (type III) endoleak (P =.001), graft migration (P =.001), and postoperative kinking of the endograft (P =.001). Forty-one patients underwent late conversion to open repair with a perioperative mortality rate of 24.4% (10/41). The cumulative rate (risk) of late conversion was approximately 2.1% per year. Risk factors (indications) for late conversion were proximal type I endoleak (P =. 001), midgraft (type III) endoleak (P =.001), type II endoleak (P =. 003), graft migration (P =.001), graft kinking (P =.001), and distal type I endoleak (P =.001). CONCLUSIONS: Endovascular repair of infrarenal aortic aneurysms with the first- and second-generation devices that predominated in this study was associated with a risk of late failure, according to an analysis of observed hard end points of 3% per year. Action taken to address the risk factors identified by the study may improve results in the future. PMID- 11013039 TI - Reducing the risk of carotid surgery: a 7-year audit of the role of monitoring and quality control assessment. AB - BACKGROUND AND PURPOSE: The current risk of stroke after carotid endarterectomy may be worse than reported in the international trials. Because studies have suggested that most operative strokes follow surgeon error, the aim of the current study was to audit the impact of introducing a strategy of perioperative monitoring and quality control assessment on outcome. METHODS: A total of 500 patients underwent carotid endarterectomy with intraoperative transcranial Doppler scan monitoring, completion angioscopy, and 3 hours of postoperative transcranial Doppler scan monitoring. The last of these guided selective dextran therapy in patients with high rates of postoperative embolization, which in previous series has been shown to be highly predictive of progression to thromboembolic stroke. RESULTS: Intimal flaps were repaired in 3% of patients and luminal thrombus removed in 4% of patients. The rate of intraoperative stroke was 0.2%. A total of 313 patients had more than one embolus detected postoperatively (96% within 2 hours of flow restoration), but only 22 patients had sustained embolization requiring dextran. Embolization ceased in all but one patient receiving dextran, although the dose had to be increased in seven patients (36%). One patient was unable to receive adequate dextran therapy because of severe cardiac failure. Overall, the 30-day death/stroke rate was 2.2%, no patient had a perioperative stroke because of carotid thrombosis, and the rate of ipsilateral embolic stroke was 0.8%. Most complications resulted from cardiac pathology or intracranial hemorrhage. CONCLUSIONS: A program of monitoring and quality control assessment has been associated with a 60% decrease in the operative risk in comparison with that observed before implementation of the protocol. PMID- 11013040 TI - Stent placement for treatment of central and peripheral venous obstruction: a long-term multi-institutional experience. AB - PURPOSE: The clinical success and patency of central and peripheral venous stents in patients with symptomatic venous obstruction (SVO) were assessed. METHODS: The records of patients with SVO treated with venous stents from 1992 to 1999 were reviewed. Demographic and procedural variables were analyzed to determine their effect on clinical success, primary patency, and secondary patency. Patency was determined by means of a follow-up duplex scan or venogram. RESULTS: Forty central venous (CV) and 14 peripheral venous (PV) obstructions were treated in 49 patients. Sixty-five stents were placed (50 CV and 15 PV), 54 in previously unstented lesions and 11 in previously stented lesions. Causes of CV lesions included catheter placement (82%), tumor compression (6%), arteriovenous fistula (AVF) and no prior catheter (2%), and other (10%). All PV lesions resulted from complications of dialysis. Indications for CV stents included limb edema (46%), AVF malfunction (30%), both limb edema and AVF malfunction (14%), and other (10%). PV stent indications were AVF malfunction (86%) and limb edema (14%). Thirteen CV stents indicated to treat tumor compression (three cases), May Thurner syndrome (one case), deep venous thrombosis (three cases), superior vena cava syndrome (one case), and lower-extremity catheter-related lesions (five cases) were excluded from the analysis of clinical outcome. Fifty-two stents (37 CV and 15 PV) were included in the analysis of clinical outcome. All CV lesions included in the analysis were complications of prolonged catheterization. Eighty nine percent of patients had end-stage renal disease and an AVF. Complications developed in 26% of patients with PV stents and in no patients with CV stents (P <.002). The mean follow-up period was 16 months. Sixty-two percent of patients required a reintervention for recurrent SVO. Only 32% of the interventions resulted in sustained symptomatic improvement. For CV stents, the primary patency rate was 85%, 27%, and 9% at 3, 12, and 24 months, respectively; the secondary patency rate was 91%, 71%, and 39% at 3, 12, and 24 months, respectively; and the clinical success rate was 94%, 94%, and 79%, at 3, 12, and 24 months, respectively. For PV stents, the primary patency rate was 73%, 17% and 17% at 3, 12, and 24 months, respectively; the secondary patency rate was 80%, 56%, and 35% at 3, 12, and 24 months, respectively; and the clinical success rate was 92%, 75%, and 42% at 3, 12 and 24 months, respectively. CONCLUSION: Stents provide a temporary benefit in most patients with central or peripheral upper-extremity SVO. Regular follow-up and reinterventions are required to maintain patency and achieve long-term clinical success. Stents used for CV lesions have higher clinical success rates than stents used for PV lesions. Patients with a reasonable life expectancy or who are unable to return for subsequent procedures should be considered for undergoing alternative therapy. PMID- 11013041 TI - Percutaneous endovascular repair of infrarenal abdominal aortic aneurysms: a feasibility study. AB - BACKGROUND: Endovascular grafting has markedly reduced the invasiveness of the treatment of abdominal aortic aneurysms. By using a modification of technique for available closure devices, we have been able to achieve percutaneous repair of aneurysms. This study reviewed our initial experience with this technique. METHODS: Demographics and background data from patients undergoing endovascular repair of abdominal aortic aneurysms were reviewed from prospectively collected registry data. Operative notes and angiographic and computed tomography scan data were retrospectively reviewed to assess the success of the percutaneous approach. RESULTS: Fourteen patients have undergone percutaneous placement of the AneuRx (Medtronic, Sunnyvale, Calif) endovascular graft, with a modification of the technique for the Prostar (Perclose, Redwood City, Calif) device for access site closure. Main graft body introduction with a 22F sheath proved successful in nine of 12 (75%) deployments. Contralateral limb deployment through a 16F sheath was successful in 10 of 14 deployments (71.4%). Reasons for conversion to open groin incisions include inadequate percutaneous hemostasis (six cases), iliofemoral dissection (four cases), device failure (one case), and compromised distal flow (one case). Percutaneous deployment success appears to be improved with larger iliac artery dimensions, decreased calcification, and limited tortuosity, because of the limitation of complications related to delivering a larger diameter sheath. Of the 13 percutaneous endograft insertions that were attempted, six (46.2%) were completely successful. CONCLUSION: Percutaneous deployment of available devices is technically feasible by using modifications of technique with percutaneous closure devices, despite large introducer sizes. Further experience with this technique offers the potential for identifying patients in whom this will prove successful and for even further reducing hospital stay and recovery times for aneurysm repair. PMID- 11013042 TI - Relationship between preoperative patency of the inferior mesenteric artery and subsequent occurrence of type II endoleak in patients undergoing endovascular repair of abdominal aortic aneurysms. AB - OBJECTIVES: The purpose of this study was (1) to find out whether preoperative inferior mesenteric artery (IMA) patency (on radiographic imaging) predicts IMA related endoleaks after endovascular repair of infrarenal abdominal aortic aneurysms, (2) to determine feasibility of measuring aneurysm sac pressures in patients with endoleaks, and (3) to report early evidence of effective endovascular obliteration of IMA endoleaks. METHODS: We studied 76 consecutive cases of infrarenal aortic aneurysms that were repaired with an endovascular approach (March 1998-April 1999). RESULTS: There were 13 (17%) endoleaks persistent 30 days after the procedure. Eleven (85%) of these 13 were IMA-related endoleaks, which were documented with selective superior mesenteric artery angiography. The preoperative finding (on computed tomographic scan) of a patent IMA does not always predict an IMA-related endoleak, but results in a statistically and clinically significant higher ratio of patients with IMA related endoleaks in the immediate postoperative period (24% versus 3%, P <.035). In eight of the 11 patients with persistent IMA-related endoleaks, measurement of intra-aneurysm sac pressures was possible, and six of these patients had systemic pressures within the excluded aneurysm sac. Nine (82%) of 11 IMA-related endoleaks were successfully obliterated by means of selective IMA embolization. CONCLUSIONS: Many endoleaks are caused by a patent IMA, and this can result in persistence of systemic pressure within the aneurysm sac. The preoperative finding (on computed tomographic scan) of a patent IMA is a predictor of increased rates of IMA endoleaks, and IMA endoleaks can be successfully obliterated through endovascular procedures, after endovascular abdominal aortic aneurysm repair. PMID- 11013043 TI - Endothelial cell response to different mechanical forces. AB - PURPOSE: Endothelial cells (ECs) are subjected to the physical forces induced by blood flow. The aim of this study was to directly compare the EC signaling pathway in response to cyclic strain and shear stress in cultured bovine aortic ECs. MATERIALS AND METHODS: The ECs were seeded on flexible collagen I-coated silicone membranes to examine the effect of cyclic strain. The membranes were deformed with a 150-mm Hg vacuum at a rate of 60 cycle/min for up to 120 minutes. For a comparison of the effect of shear stress, ECs from the same batch as used in the strain experiments were seeded on collagen I-coated silicone sheets. The ECs were then subjected to 10 dyne/cm(2) shear with the use of a parallel flow chamber for up to 120 minutes. Activation of the mitogen- activated protein kinases was assessed by determining phosphorylation of extracellular signal regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38 with immunoblotting. RESULTS: ERK, JNK, and p38 were activated by both cyclic strain and shear stress. Both cyclic strain and shear stress activated JNK with a similar temporal pattern and magnitude and a peak at 30 minutes. However, shear stress induced a more robust and rapid activation of ERK and p38, compared with cyclic strain. CONCLUSIONS: Our results indicate that different mechanical forces induced differential activation of mitogen-activated protein kinases. This suggests that there may be different mechanoreceptors in ECs to detect the different forces or alternative coupling pathways from a single receptor. PMID- 11013044 TI - Pulsatile wall motion and blood pressure in aneurysms with open and thrombosed endoleaks--comparison of a wall track system and M-mode ultrasound scanning: an in vitro and animal study. AB - OBJECTIVE: Pulsatile wall motion has been suggested as a means by which to evaluate abdominal aortic aneurysms after exclusion from the circulation to determine whether the treatment has been effective. The objective of this study was to investigate the relations between pulsatile wall motion and both the mean and pulse pressures within the aneurysmal sac for both patent and thrombosed endoleaks. Furthermore, we compared the measurements of pulsatile wall motion by means of M-mode ultrasound scanning and a wall track system to determine the most reliable technique. METHODS: First, interobserver and intraobserver variability of M-mode ultrasound scan measurements was determined at different pressure levels in a cow iliac artery placed in an in vitro circulation. M-mode ultrasound scanning and a wall track system were compared in the same model. Second, in an animal experiment, an aneurysm and endoleak model with both patent and thrombosed endoleaks was created. Systemic and aneurysmal mean and pulse pressures were recorded synchronically with pulsatile wall motion by means of M-mode ultrasound scanning and a wall track system. RESULTS: The intraobserver and interobserver variability values for M-mode ultrasound scan measurement in vitro were 0.11 mm (SD = 0.10 mm) and 0.15 mm (SD = 0.13 mm), respectively. In the animal study, a significant difference existed with respect to the level of pulse pressure within the aneurysmal sac between the group with pulsatile wall motion and the group without such motion (P <.0001). The presence of pulsatile wall motion was not correlated with the level of aneurysmal mean pressure. The level of pulsatile wall motion determined by means of M-mode ultrasound scanning correlated well with the level determined by means of the wall track system (r = 0. 74; P =.01). For the level of pulsatile wall motion determined by means of M-mode ultrasound scanning, a significant difference between patent and thrombosed endoleaks existed (P =.04). For detecting endoleaks, the sensitivity and specificity of pulsatile wall motion as determined by means of the wall track system were 52% and 100%, respectively, and the sensitivity and specificity of pulsatile wall motion as determined by means of M-mode ultrasound scanning were 64% and 67%, respectively. For the detection of pulse pressure in the aneurysmal sac, the sensitivity and specificity of pulsatile wall motion as determined by means of the wall track system were 76% and 100%, respectively, and the sensitivity and specificity of pulsatile wall motion as determined by means of M-mode ultrasound scanning were 90% and 71%, respectively. CONCLUSIONS: We found that pulsatile wall motion is correlated with aneurysmal pulse pressure but not with the mean level of pressure inside the aneurysm. Although measurements of pulsatile wall motion are of great theoretic value when groups of patients who have undergone endovascular aneurysm repair are being compared, this method appears to be unreliable in a clinical setting with respect to determining whether the aneurysmal sac is still pressurized in individual patients. PMID- 11013045 TI - Recombinant thrombomodulin inhibits arterial smooth muscle cell proliferation induced by thrombin. AB - PURPOSE: Restenosis after angioplasty or bypass grafting to restore circulation to ischemic organs is still an unsolved problem. Thrombin generated in high concentrations at the sites of vascular injury plays a central role in thrombosis and hemostasis. alpha-Thrombin has also been implicated as a mitogen for smooth muscle cell (SMC) proliferation that contributes to arterial restenosis. Thrombomodulin has a high affinity of binding with thrombin and converts thrombin from a procoagulant to an anticoagulant. This study was designed to examine whether thrombomodulin could also moderate the thrombin-mediated SMC proliferative response. METHODS: Porcine carotid artery SMCs (passages 4-7) were plated onto 96-well plates and incubated for 3 days. After growth arrest in a defined serum-free medium for 2 to 3 days, SMCs were subjected to the reagents as follows: (1) human alpha-thrombin, (2) recombinant human soluble thrombomodulin containing a chondroitin sulfate moiety, (3) thrombin receptor agonist peptide (SFLLRNPNDKYEPF), and (4) alpha-thrombin or thrombin receptor agonist peptide combined with recombinant thrombomodulin (rTM). The viability and proliferation status of SMCs were quantified with MTT (thiazolyl blue) mitochondrial function and bromodeoxyuridine (BrdU)-DNA incorporation assays. RESULTS: Human alpha thrombin increased SMC proliferation in a dose dependent manner by more than 25% and 30% with thrombin 1 U/mL to 3 U/mL compared with control groups on day 7 (P <.006). rTM concentrations from 0.5 microg/mL to 3 microg/mL have no significant effect on SMC growth. The stimulation of SMC proliferation induced by alpha thrombin at 0.5 U/mL, 1 U/mL, and 2 U/mL was significantly inhibited with rTM at 2 microg/mL and 3 microg/mL on days 3, 7, and 10 as evaluated with MTT assay (P <.01 to <.05) and BrdU-DNA incorporation assay on day 3 (P <.008). Thrombin receptor agonist peptide increased SMC BrdU-DNA incorporation at 48 hours (P <.007), and its effect was not altered by rTM. CONCLUSION: rTM containing all of the extracellular domains of thrombomodulin inhibits the effect of thrombin on SMC proliferation in vitro. Because thrombin is a mitogenic mediator of SMC in vascular injury, inhibition of its function in vivo could help to prevent SMC hyperplasia. The success of further studies in vivo may lead to use of rTM for decreasing or preventing arterial restenosis. PMID- 11013047 TI - Bypass graft to the contralateral internal jugular vein for venous outflow obstruction of a functioning hemodialysis access fistula. AB - As survival among patients with renal failure improves, vascular access becomes more difficult, and preservation of functioning access increases in importance. Subclavian vein thrombosis associated with a distal arteriovenous fistula can result in massive and debilitating swelling of the affected extremity. We describe a novel crossover bypass grafting procedure to the contralateral internal jugular vein in a patient with a thrombosed internal jugular and subclavian vein. This procedure resulted in preservation of the functioning arteriovenous fistula and resolution of the symptoms. Unlike previously described crossover procedures to the contralateral basilic or axillary veins, this bypass graft has the added benefit of not obviating future fistula creation in that extremity. PMID- 11013046 TI - Laparoscopic release of celiac artery compression syndrome facilitated by laparoscopic ultrasound scanning to confirm restoration of flow. AB - A 43-year-old woman presented with symptomatic mesenteric ischemia caused by median arcuate ligament compression of her celiac artery. Magnetic resonance angiography clearly demonstrated stenosis of the proximal celiac artery. She underwent laparoscopic decompression by division of the ligament and excision of the celiac plexus. Laparoscopic Doppler ultrasound scanning demonstrated markedly improved flow in the artery. She was discharged in 15 hours and reported complete resolution of her symptoms at the 3-month postoperative visit. Laparoscopy provides a less invasive but equally effective method for decompressing the celiac artery as well as assessing adequacy of flow after its release. PMID- 11013048 TI - Malignant carotid body tumor: a case report. AB - Carotid body tumors (CBTs) have an unpredictable history with no correlation between histology and clinical behavior. Of reported cases since 1891, local and distant metastases appear in approximately 10% of cases and remain the hallmark of malignancy. Currently, there are not enough data to support a single treatment regimen for malignant CBTs. The reported case demonstrates some unanswered issues with regard to malignant CBTs to include lymph node dissection, the need for carotid resection, and the role of radiation therapy. A 46-year-old pathologist underwent a resection of a Shamblin I CBT, to include jugular lymph node sampling, without complication. There was lymph node involvement, and tumor cells were found on the margins of the pathologic specimen. Subsequent carotid resection with reversed interposition saphenous vein graft and modified neck dissection were performed again without complication. Follow-up at 4 years has been uneventful. Diagnosis of CBTs with the use of magnetic resonance angiography, magnetic resonance imaging, color flow duplex scanning, and the role of arteriography are reviewed. The current treatment options are discussed with reference to primary lymph node sampling, carotid resection, and neck dissection in malignant cases. This case demonstrates that the unpredictable nature of CBTs and their malignant potential warrant aggressive initial local treatment to include jugular lymph node sampling and complete tumor resection. PMID- 11013050 TI - The distinguished service award for the Society for Vascular Surgery, 2000: D. Emerick Szilagyi, MD. PMID- 11013049 TI - Ruptured internal carotid aneurysm resulting from neurofibromatosis: treatment with intraluminal stent graft. AB - PURPOSE: This report shows a method of treatment for life-threatening hemorrhage due to rupture of an aneurysm in the cervical internal carotid artery caused by neurofibromatosis. METHODS: Ten days after delivery of healthy twins, a 28-year old woman with known neurofibromatosis had sudden massive swelling in the left neck. After initial tracheostomy, angiography confirmed rupture of the mid cervical internal carotid artery as well as contribution to the resultant pseudoaneurysm from external carotid branches. Treatment began with coil embolization of the external carotid branches. The internal carotid lesion, a defect approximately 1 cm in length, was then closed through use of two stent grafts, each made from Palmaz stents and 3-mm polytetrafluorethylene grafts predilated to 6 mm. The neck hematoma was then evacuated surgically. RESULTS: Completion angiography and computed tomographic scanning confirmed control of the hemorrhage. The patient survived neurologically intact with the exception of cranial nerve deficits caused by the hemorrhage. The tracheostomy tube was removed 3 weeks postoperatively. Follow-up computed tomographic scanning showed a gradual decrease in the size of the cervical soft tissue and no recurrent aneurysm. CONCLUSION: Neurofibromatosis is a rare cause of aneurysmal degeneration of blood vessels. Repair of a ruptured cervical internal carotid artery aneurysm, though feasible, is difficult with stent grafts; however, this is a better option than surgical intervention in inaccessible vessels. PMID- 11013051 TI - Neuromusculoskeletal conditions of the upper extremity: are they due to repetitive occupational trauma? AB - The significance of neuromusculoskeletal conditions in the workplace is the subject of much discussion among occupational medicine professionals. There are differing philosophies as to what constitutes appropriate diagnostic criteria for identification of these conditions. The traditional diagnostic model requires the presence of objective pathology. An emerging symptom-based model accepts that symptoms by themselves can constitute a diagnostic entity. The extent to which these conditions are considered to be associated with occupational activity depends greatly upon which of the two models is employed. This chapter presents an overview of each diagnostic model and a discussion of the impact each has on the prevalence of identified conditions and the manner in which the various diagnostic requirements can affect treatment, prevention, and disability rating protocols. PMID- 11013052 TI - S.P.I.C.E.--a model for reducing the incidence and costs of occupationally entitled claims. AB - A review of national statistics and recent studies strongly suggests that current administrative and medical systems, when applied to managing workers' compensation claims and other disability-related benefit programs, are often ineffective and costly, and can even promote disability. With numerous medical and occupational health articles published daily, it is difficult to develop practical strategies for daily management of disability claims that make use of current information. It is the authors' view that a comprehensive, dynamic model for management exists within the military's "Forward Treatment" methodology. This model, originally published in The Journal of Occupational Rehabilitation, has been expanded to include those methods demonstrated in literature to both reduce claims and deal with them in an efficient, fair, and timely manner. Because military personnel and employed individuals are similarly entitled, the military's proven model can be effective in reducing claim rates and costs associated with workers' compensation as well as short- and long-term disability programs. The model, given the acronym S.P. I.C.E., includes five components: Simplicity, Proximity, Immediacy, Centrality, and Expectancy. PMID- 11013053 TI - Unified fitness report for the workplace. AB - Fitness statements often are required of physicians by patients, employers, governmental agencies, and insurance providers to determine if the patient is fit for duty. Physicians making these ability statements are legally obligated to carefully justify them when placing or excluding individuals from the workplace. The Americans with Disabilities Act (ADA) mandates that medical providers use justifiable criteria and rational thought when determining the capability and risk of an individual. This chapter reviews the legal requirements of the ADA for employers and physicians and presents a uniform methodology that both can use to determine the performance capability of an individual with a temporary or permanent impairment or disability. PMID- 11013054 TI - The importance of illness behavior in disability management. AB - Abnormal illness behaviors, ranging from non-deliberate distortion to intentional deception, are associated with clinical phenomena that lie along a continuum from unconscious symptom exaggeration to psychiatric disorders and malingering. Failure to recognize abnormal illness behavior leads to inappropriate treatment and erroneous estimates of impairment or disability. This review is divided into three sections. First, basic terms are defined, including dissimulation, distortion, deception, misattribution, false imputation, and malingering. Second, syndromes characterized by abnormal illness behavior are described, including somatization, somatoform disorders, factitious disorders, and symptom magnification. Third, methods for detecting deception are illustrated, including maximum voluntary effort assessment, objective personality inventories, and symptom validity testing. PMID- 11013055 TI - Management of chronic pain and control of long-term disability. AB - Chronic pain has become a major public health problem. Often, the availability of entitlement programs as well as psychosocial, occupational, and other nonmedical factors-rather than objective pathophysiology-are major contributors to disability. In this chapter, the authors discuss the relationship between impairment and disability and detail factors likely to predict or contribute to adverse clinical outcome and disability. Guidelines for disability prevention also are examined. PMID- 11013056 TI - Assessment and management of upper and lower extremity impairment and disability. AB - The assessment and management of occupational and nonoccupational injuries and illnesses of the extremities require knowledge of the same concepts of impairment and disability that are used when evaluating or treating problems of other body systems. Dr. Talmage discusses the need for an accurate and objectively verifiable diagnosis and focuses on upper extremity pain in workers and the term tendinitis'. The concept of maximal medical improvement is considered, with examples of knee injuries to the menisci and localized articular cartilage defects. Causation controversies are explored using carpal tunnel syndrome as the example. The impairment rating process is illustrated by use of the AMA Guides system. A "checklist" to insure completeness of the examination of extremity problems is offered. Disability assessment and management is discussed, with an emphasis on the difference between abilities (capacities) and risk-based restrictions. The author concludes with thoughts on how to minimize disability. PMID- 11013057 TI - Worksite disability management model for effective return-to-work planning. AB - The growth of disability management programs represents a paradigm shift from traditional clinic-based rehabilitation services to worksite-based interventions that dramatically reduce lost time and costs. Supportive policies and steps in the return-to-work process are illustrated, from the point of worker injury and early intervention, through work ability assessment, return-to-work planning, job site accommodation, and successful return to work. Creative return-to-work options, including "job banks," are discussed as practical methods to facilitate the worker's gradual return to full duty status while completing the medical recovery process. Worker, worksite, and community resource factors related to return-to-work outcomes are discussed. PMID- 11013059 TI - Use of functional employment testing to facilitate safe job placement. AB - Functional testing has evolved to a new sophistication, and is currently used in a variety of situations to assist the employer and physician in safely placing an individual at the job site. The functional capacity evaluation can be employed in several ways, not only to place individuals safely in jobs, but also to monitor their progress throughout recovery from an injury or illness and aid in the establishment of vocational counseling and planning. Many legal issues now alter employer techniques for hiring and assigning people to jobs. This state of affairs places increasing importance on functional testing. PMID- 11013058 TI - Psychosocial factors and risk of pain and disability. AB - This article reviews the research on the risk of pain and disability due to psychosocial variables. Variables such as general distress, psychopathology, depression, abuse, and catastrophizing are discussed in relation to the risk of disability. Ways to conceptualize the complex relationships among pain, disability, and several psychosocial variables are also explored. In addition, the identification of adaptive and of protective ways to manage pain and decrease the risk of disability is highlighted. Finally, the authors recommend areas for future research. PMID- 11013060 TI - To smile again. Reanimation for unilateral facial palsy. PMID- 11013061 TI - Organ donation. 3. Brain stem death. AB - Following an exploration of nurses' perceptions of organ donation, and of consent and patient's rights in the preceding articles, this series concludes with an examination of brain stem death, and in particular the literature which challenges popular assumptions about it. This makes challenging and sometimes disturbing reading, but as the author reminds us, these are issues which theatre nurses cannot ignore. PMID- 11013062 TI - Lifeskills training. You've read the books ... now it's time to apply it to life! PMID- 11013063 TI - Horizontal violence in the operating room. PMID- 11013064 TI - Concepts of health and the health promoting role of the perioperative nurse. AB - Health promotion is an important part of the work of a wide range of healthcare professionals. The United Kingdom Central Council (UKCC 1983) states that education is a statutory responsibility and one of the most important and challenging aspects of the nurse's role. The nurse has a professional as well as a moral responsibility to educate for health as well as to care for the sick (UKCC 1994). PMID- 11013065 TI - A framework for perioperative advanced practice. PMID- 11013066 TI - Breathing circuits and their uses. PMID- 11013067 TI - Estrogen-induced remodeling of hypothalamic neural circuitry. AB - For decades, sexual behavior has been a valuable model system for behavioral neuroscientists studying the neural basis of motivated behaviors. One striking example of a change in motivation is the binary switch in sexual receptivity that occurs during the estrous cycle in female rats. Investigations of the neural basis of this change in behavior have fundamentally advanced our understanding of both behaviorally relevant neural pathways and basic mechanisms of steroid action in the brain. These advances have made this behavioral model system a staple of neuroendocrinology. A challenge that remains before us, given our current understanding of the circuitry and chemistry, is to develop a coherent model of how neural plasticity in the hypothalamus contributes to the dependence of this behavior on motivational state. This review will focus on the ventromedial nucleus of the hypothalamus, especially its ventrolateral subdivision. First, the anatomical, neurochemical, and functional aspects of the macro- and microcircuitry of this brain region will be discussed, followed by a discussion of the likely mechanisms of estrogen action within the ventrolateral VMH. Then, the evidence for estrogen-induced neural plasticity will be considered, including a comparison with the effects of estrogen on synaptic organization in other brain regions. Finally, a working model of neural plasticity within the ventrolateral VMH microcircuitry will be presented as a starting point for future experiments to verify or, more likely, revise and expand. PMID- 11013068 TI - Growth hormone in the brain: characteristics of specific brain targets for the hormone and their functional significance. AB - During the past decade studies have shown that growth hormone (GH) may exert profound effects on the central nervous system (CNS). For instance, GH replacement therapy was found to improve the psychological capabilities in adult GH deficient (GHD) patients. Furthermore, beneficial effects of the hormone on certain functions, including memory, mental alertness, motivation, and working capacity, have been reported. Likewise, GH treatment of GHD children has been observed to produce significant improvement in many behavioral problems seen in these individuals. Studies also indicated that GH therapy affects the cerebrospinal fluid levels of various hormones and neurotransmitters. Further support that the CNS is a target for GH emerges from observations indicating that the hormone may cross the blood-brain barrier (BBB) and from studies confirming the presence of GH receptors in the brain. It was previously shown that specific binding sites for GH are present in discrete areas in the CNS of both humans and rats. Among these regions are the choroid plexus, hippocampus, hypothalamus, and spinal cord. The density of GH binding in the various brain regions was found to decline with increasing age. More recently, we were able to clone and determine the structure of several GH receptors in the rat and human brain. Although the brain receptor proteins for the hormone were shown to differ in molecular size compared to those present in peripheral tissues the corresponding transcripts did not seem to differ from their peripheral congeners. GH receptors in the hypothalamus are likely to be involved in the regulatory mechanism for hormone secretion and those located in the choroid plexus have been suggested to have a role in the receptor-mediated transport of GH across the BBB. The functions mediated by the GH receptors identified in the hippocampus are not yet known but recently it was speculated that they may be involved in the hormone's action on memory and cognitive functions. PMID- 11013069 TI - New members of the parathyroid hormone/parathyroid hormone receptor family: the parathyroid hormone 2 receptor and tuberoinfundibular peptide of 39 residues. AB - The parathyroid hormone (PTH) family currently includes three peptides and three receptors. PTH regulates calcium homeostasis through bone and kidney PTH1 receptors. PTH-related peptide, probably also through PTH1 receptors, regulates skeletal, pancreatic, epidermal, and mammary gland differentiation and bladder and vascular smooth muscle relaxation and has a CNS role that is under investigation. Tuberoinfundibular peptide of 39 residues (TIP39) was recently purified from bovine hypothalamus based on selective PTH2 receptor activation. PTH2 receptor expression is greatest in the CNS, where it is concentrated in limbic, hypothalamic, and sensory areas, especially hypothalamic periventricular neurons, nerve terminals in the median eminence, superficial layers of the spinal cord dorsal horn, and the caudal part of the sensory trigeminal nucleus. It is also present in a number of endocrine cells. Thus TIP39 and PTH2 receptor influenced functions may range from pituitary and pancreatic hormone release to pain perception. A third PTH-recognizing receptor has been found in zebrafish. PMID- 11013070 TI - Fine-scale comparative mapping of the human 7q11.23 region and the orthologous region on mouse chromosome 5G: the low-copy repeats that flank the Williams Beuren syndrome deletion arose at breakpoint sites of an evolutionary inversion(s). AB - Williams-Beuren syndrome (WBS) is a developmental disorder caused by haploinsufficiency for genes deleted in chromosome band 7q11.23. A common deletion including at least 16-17 genes has been defined in the great majority of patients. We have completed a physical and transcription map of the WBS region based on analysis of high-throughput genome sequence data and assembly of a BAC/PAC/YAC contig, including the characterization of large blocks of gene containing low-copy-number repeat elements that flank the commonly deleted interval. The WBS deletions arise as a consequence of unequal crossing over between these highly homologous sequences, which confer susceptibility to local chromosome rearrangements. We have also completed a clone contig, genetic, and long-range restriction map of the mouse homologous region, including the orthologues of all identified genes in the human map. The order of the intradeletion genes appears to be conserved in mouse, and no low-copy-number repeats are found in the region. However, the deletion region is inverted relative to the human map, exactly at the flanking regions. Thus, we have identified an evolutionary inversion with chromosomal breakpoints at the sites where the human 7q11.23 low-copy-number repeats are located. Additional comparative mapping suggests a model for human chromosome 7 evolution due to serial inversions leading to genomic duplications. This high-resolution mouse map provides the framework required for the generation of mouse models for WBS mimicking the human molecular defect. PMID- 11013071 TI - Human-chimpanzee DNA sequence variation in the four major genes of the renin angiotensin system. AB - The renin angiotensin system (RAS) is involved in blood pressure control and water/sodium metabolism. The genes encoding the proteins of this system are candidate genes for essential hypertension. The RAS involves four main molecules: angiotensinogen, renin, angiotensin I-converting enzyme, and the angiotensin II type 1 receptor (encoded by the genes AGT, REN, DCP1, and AGTR1, respectively). We performed a molecular screening over 17,037 bp of the coding and 5' and 3' untranslated regions of these genes, from three to six common chimpanzees. We identified 44 single-nucleotide polymorphisms (SNPs) in chimpanzee samples, including 18 coding-region SNPs, 5 of which led to an amino acid replacement. We observed common and different features at various sites (synonymous, nonsynonymous, and noncoding) within and between the four chimpanzee genes: (1) the nucleotide diversity at noncoding sites was similar; (2) the nucleotide diversity at nonsynonymous sites was low, probably reflecting purifying selection, except for the AGT gene; (3) the nucleotide diversity at synonymous sites, which was dependent on the G+C content at the third position of the codon, was high, except for the AGTR1 gene. Comparison of the chimpanzee SNPs with those previously reported for humans identified 119 sites with fixed differences (including 62 coding sites, 17 of which resulted in amino acid differences between the species). Analysis of polymorphism within species and divergence between species shed light on the evolutionary constraints on these genes. In particular, comparison of the pattern of mutation at polymorphic and fixed sites between humans and chimpanzees suggested that the high G+C content of the DCP1 gene was maintained by positive selection at its silent sites. Finally, we propose 68 ancestral alleles for the human RAS genes and discuss the implications for their use in future hypertension-susceptibility association studies. PMID- 11013072 TI - Cross-referencing radiation hybrid data to the recombination map: lessons from mouse chromosome 18. AB - We are building a framework map of known-order anchor markers between the mouse T31 radiation hybrid (RH) panel and the recombination map based on The Jackson Laboratory (TJL) interspecific backcross panels using the established genetic order to evaluate and strengthen the RH results. In making this map comparison, we have elucidated several problems inherent in RH mapping and minimized these by careful attention to data gathering and interpretation methods. We describe lessons and pitfalls of developing radiation hybrid maps, using the example of mouse Chromosome 18, for which we have built a framework map of microsatellite anchor loci spanning the entire chromosome at significant LOD with no gaps. Sixty five D18Mit- simple sequence length polymorphism (SSLP) markers form a continuous linkage along the T31 RH Chromosome 18 (RH map length 1598 cR, genetic length 41 cM) with all LODs greater than 6. These markers are also placed on TJL interspecific backcrosses, and the order of the markers in the two systems is in complete agreement. We are continuing to cross-reference the RH data to TJL backcross data for the other mouse chromosomes to improve further the power of RH mapping and to integrate more precisely the extensive existing recombination mapping data for the mouse with the incoming radiation hybrid map data. PMID- 11013073 TI - The characterization of the common fragile site FRA16D and its involvement in multiple myeloma translocations. AB - Fragile sites appear as breaks, gaps, or decondensations on metaphase chromosomes when cells are grown under specific culture conditions. The breaks are nonrandom, appearing in defined, conserved locations throughout the mammalian genome. Common fragile sites, as their name implies, are present in virtually all individuals. With three common fragile sites cloned, their mechanism of expression and the role, if any, they play in human disease are still unclear. We have assembled a BAC contig of >1 Mb across the second most active common fragile site, FRA16D (16q23.2). We fluorescently labeled these BACs and used them as probes on metaphases from aphidicolin-induced lymphocytes and demonstrated that FRA16D decondensation/breakage occurs over a region of at least 1 Mb. Thus, this is the largest common fragile site cloned to date. Microsatellite markers that map within FRA16D show a very high loss in prostate, breast, and ovarian tumors, indicating that loss within this fragile site may be important in the development or progression of these tumors. In addition, a common t(14q32;16q23) translocation is observed in up to 25% of all multiple myelomas (MM). We localized four of four such cloned t(14;16) MM breakpoints within the FRA16D region. This work further demonstrates that the common fragile sites may play an important role in cancer development. PMID- 11013074 TI - Quantitative trait loci affecting prion incubation time in mice. AB - Although the gene encoding prion protein (PrP) is the major determinant of susceptibility to prion disease, other genes also affect prion incubation time in mice and may be involved in prion replication. Scrapie incubation time was analyzed as a quantitative trait using crosses between SJL/J and CAST/Ei mice; these mouse strains encode identical PrP molecules but have different incubation periods. Our analysis revealed loci on Chromosomes 9 and 11 that affect prion susceptibility. PMID- 11013075 TI - Identification and structural analysis of human RBM8A and RBM8B: two highly conserved RNA-binding motif proteins that interact with OVCA1, a candidate tumor suppressor. AB - The OVCA1 gene is a candidate for the breast and ovarian tumor suppressor gene at chromosome 17p13.3. To help determine the function(s) of OVCA1, we used a yeast two-hybrid screening approach to identify OVCA1-associating proteins. One such protein, which we initially referred to as BOV-1 (binder of OVCA1-1) is 173 or 174 amino acids in length and appears to be a new member of a highly conserved RNA-binding motif (RBM) protein family that is highly conserved evolutionarily. Northern blot analysis revealed that BOV-1 is ubiquitously expressed and that three distinct messenger RNA species are expressed, 1-, 3.2-, and 5.8-kb transcripts. The 1-kb transcript is the most abundant and is expressed at high levels in the testis, heart, placenta, spleen, thymus, and lymphocytes. Using fluorescence in situ hybridization and the 5.8-kb complementary DNA probe, we determined that BOV-1 maps to both chromosome 5q13-q14 and chromosome 14q22-q23. Further sequence analysis determined that the gene coding the 1- and the 3.2-kb transcripts (HGMW-approved gene symbol RBM8A) maps to 14q22-q23, whereas a second highly related gene coding for the 5.8-kb transcript resides at chromosome 5q13 q14 (HGMW-approved gene symbol RBM8B). The predicted proteins encoded by RBM8A and RBM8B are identical except that RBM8B is 16 amino acids shorter at its N terminus. Molecular modeling of the RNA-binding domain of RBM8A and RBM8B, based on homology to the sex-lethal protein of Drosophila, identifies conserved residues in the RBM8 protein family that are likely to contact RNA in a protein RNA complex. The conservation of sequence and structure through such an evolutionarily divergent group of organisms suggests an important function for the RBM8 family of proteins. PMID- 11013077 TI - Identification and characterization of the miniature pig Huntington's disease gene homolog: evidence for conservation and polymorphism in the CAG triplet repeat. AB - Huntington's disease (HD) is associated with a significant expansion of a CAG trinucleotide repeat, which results in a lengthened polyglutamine tract in the single gene product, huntingtin, on human 4p16.3. We isolated cDNA clones that encompassed the entire coding sequence of the miniature pig HD gene (Sus HD) from two porcine testis cDNA libraries. The cDNA contig revealed a 12,749-nucleotide transcript coding for a 345-kDa protein (3139 amino acid residues), which exhibited 96% peptide sequence homology to human huntingtin. Northern blot analysis revealed that the Sus HD gene was ubiquitously expressed as two large transcripts of approximately 11 and 13 kb in size in all the tested tissues, much like the human HD gene. The CAG trinucleotide repeat was found to be interrupted by CAA triplets and to encode 17 or 18 consecutive glutamine residues. In our laboratory stock of miniature pig, three allotypes in the triplet repeat sequence were found. Thus, the Sus HD gene closely resembles its human counterpart in terms of sequence and expression pattern. In particular, human-miniature pig similarities in the normal length of the CAG triplet repeat as well as its repeat number polymorphism may indicate that miniature pig would provide a good animal model for Huntington's disease. PMID- 11013076 TI - Identification of the human Mnk2 gene (MKNK2) through protein interaction with estrogen receptor beta. AB - We have identified and characterized the human Mnk2 gene (HGMW-approved gene symbol MKNK2) through a yeast two-hybrid screen in which the Mnk2 protein interacted with the ligand-binding domain of estrogen receptor beta (ERbeta). Human Mnk2 is homologous to murine Mnk2 ( approximately 94% identical) and human Mnk1 (71% identical), both of which encode MAP kinase interacting kinases that are phosphorylated and activated by ERK1 and 2. This report presents a thorough genomic sequence analysis revealing that the human Mnk2 gene has two C-terminal splice variants, designated here as Mnk2a and Mnk2b. These two isoforms are identical over the first 385 amino acids of the coding sequence and differ only in the final exon which encodes an additional 80 residues for Mnk2a and 29 residues for Mnk2b. A more detailed biological analysis in yeast showed that the Mnk2 interaction was selective for ERbeta as opposed to ERalpha and that the interaction was specific to Mnk2b as opposed to Mnk2a or Mnk1. This pattern was reproduced in a mammalian two-hybrid system using a completely different set of fusion partners; and in both yeast and mammalian systems, the addition of estradiol decreased the interaction. While it remains unknown whether ERbeta is a substrate of Mnk2, the interaction of these two proteins is reminiscent of ERalpha and ribosomal S6 kinase (p90-RSK), another MAP kinase-regulated kinase homologous to Mnk2 that is known to phosphorylate ERalpha. PMID- 11013078 TI - Characterization of mRAD18Sc, a mouse homolog of the yeast postreplication repair gene RAD18. AB - The RAD18 gene of the yeast Saccharomyces cerevisiae encodes a protein with ssDNA binding activity that interacts with the ubiquitin-conjugating enzyme RAD6 and plays an important role in postreplication repair. We identified and characterized the putative mouse homolog of RAD18, designated mRAD18Sc. The mRAD18Sc open reading frame encodes a 509-amino-acid polypeptide that is strongly conserved in size and sequence between yeast and mammals, with specific conservation of the RING-zinc-finger and the classic zinc-finger domain. The degree of sequence conservation between mRAD18Sc, RAD18, and homologous sequences identified in other species (NuvA from Aspergillus nidulans and Uvs-2 from Neurospora crassa) is entirely consistent with the evolutionary relationship of these organisms, strongly arguing that these genes are one another's homologs. Consistent with the presence of a nuclear translocation signal in the amino acid sequence, we observed the nuclear localization of GFP-tagged mRAD18Sc after stable transfection to HeLa cells. mRNA expression of mRAD18Sc in the mouse was observed in thymus, spleen, brain, and ovary, but was most pronounced in testis, with the highest level of expression in pachytene-stage primary spermatocytes, suggesting that mRAD18Sc plays a role in meiosis of spermatogenesis. Finally, we mapped the mRAD18Sc gene on mouse chromosome 6F. PMID- 11013079 TI - Annexin A11 (ANXA11) gene structure as the progenitor of paralogous annexins and source of orthologous cDNA isoforms. AB - The genomic organization of the annexin A11 gene was determined in mouse and human to assess its congruity with other family members and to examine the species variation in alternative splicing patterns. Mouse annexin A11 genomic clones were characterized by restriction analysis, Southern blotting, and DNA sequencing, and the homologous human gene (HGMW-approved gene symbol ANXA11) was deciphered from high-throughput genomic sequence with coanalysis of expressed sequence tags. Exons 6-15 of the tetrad core repeat region differ from annexins A7 and A13 but are spliced identically to other phylogenetic descendents, making annexin A11 the putative primary progenitor of up to nine paralogous human annexins. The 5' regions consist of untranslated exon 1, followed by an extensive intron 1 comprising almost half the total gene length of >40 kb, and additional GC-rich exons 2-5 encoding the proline- and glycine-rich amino-terminus. Distinct cDNA isoforms in cow and human were determined to be unique to each species and hence of dubious general significance for this gene's function. Multiple transcription start sites were revealed by primer extension analysis of the mouse gene, and transfection constructs containing the prospective promoter generated transcriptional activity comparable to that of the SV40 promoter. Internal repetitive elements and vicinal gene markers were mapped for the complete human annexin A11 gene sequence to characterize the surrounding genomic environment. PMID- 11013080 TI - Cloning of the genomic locus of mouse SH2 containing inositol 5-phosphatase (SHIP) and a novel 110-kDa splice isoform, SHIPdelta. AB - The SH2 domain containing inositol 5'-phosphatase (SHIP) was initially described as a 145-kDa protein phosphorylated on tyrosines upon growth factor and cytokine stimulation. It was shown to be phosphorylated after Fc and B cell receptor activation and plays a role in negative signaling. Different isoforms of the SHIP protein result from alternative mRNA splicing, proteolysis, or a combination of both. The expression of discrete SHIP isoforms changes with the potential developmental-dependent maturation state of myeloid cells, suggesting mechanisms for the regulation of SHIP interactions with other signaling molecules. A p135 (SHIPbeta) spliced isoform is known to be expressed in developing myeloid cells. Now we have identified a new SHIP isoform, SHIPdelta, which is the product of an out-of-frame splice with a deletion of 167 nucleotides in the C-terminal region, resulting in an approximately 110-kDa protein. Biochemically, SHIPdelta differs from SHIPalpha by exhibiting little or no tyrosine phosphorylation or association with the signaling protein Shc after M-CSF activation of FD-Fms cells. In addition, we have characterized the structure of the entire SHIP genomic locus, which provides a basis for understanding the alternative splicing events. SHIP is expressed in hematopoiesis and spermatogenesis, and we also describe the promoter for the SHIP gene, which has potential for explaining the tissue-specific expression pattern. PMID- 11013081 TI - Human NB-2 of the contactin subgroup molecules: chromosomal localization of the gene (CNTN5) and distinct expression pattern from other subgroup members. AB - NB-2 is one of the neural recognition molecules in the contactin subgroup, which belongs to the immunoglobulin superfamily. In rat, the six molecules in this subgroup that have been reported to date are contactin, TAG-1, BIG-1, BIG-2, NB 2, and NB-3. We have isolated cDNAs encoding the two splicing isoforms of human NB-2. The long isoform of human NB-2 consists of 1100 amino acids residues that are 91% homologous to rat NB-2 at the amino acid sequence level. The short isoform lacks 74 amino acid residues between residues 19 and 93 of the long isoform. Among various regions of the adult human brain, high-level expression of NB-2 was detected in the amygdala and occipital lobe, whereas expression was low in the corpus callosum, caudate nucleus, and spinal cord. Although there were some differences, the expression pattern of NB-2 was the most similar to that of BIG-1 in the brain. Likewise, contactin and BIG-2 exhibited similar expression patterns. The expression of TAG-1 showed the least regional differences. The human NB-2 gene (CNTN5) was mapped to chromosome 11q21-q22.2 by fluorescence in situ hybridization. Our results suggest that the NB-2 gene may contribute to human neurological disorders. PMID- 11013083 TI - Dating the origin of the V170M mutation causing non-type I cystinuria in Libyan Jews by linkage disequilibrium and physical mapping of the SLC7A9 gene. AB - Cystinuria is an autosomal recessive disorder of the transepithelial transport of amino acids, clinically manifested by the development of kidney stones. Mutations in the gene encoding rBAT (SLC3A1, on chromosome 2p16.3) are linked to type I cystinuria, while the SLC7A9 locus (19q13.1), expressing b0,+ AT protein, is involved in non-type I cystinuria, which is very common among Libyan Jews. Applying two methods for linkage disequilibrium analysis to haplotype data spanning six 19q12-q13.1 polymorphic markers, and relying on the physical distances between the markers and the recently mapped SLC7A9 (CSNU3) locus, the age of the founder missense V170M mutation causing non-type I cystinuria in Jews of Libyan ancestry is calculated to be approximately 14 to 15 generations (g) (95% confidence interval: 9-20 g) or slightly more. The estimated age dates the most recent common ancestor of the mutation-bearing chromosomes back to the time (or some decades before) Jewish families settled in Libya following their expulsion from the Iberian Peninsula. This finding makes the molecular population genetics of cystinuria understandable in the context of the Libyan Jews' history. PMID- 11013084 TI - Genetic and physical mapping of the cerebellar deficient folia (cdf) locus on mouse chromosome 6. AB - Cerebellar deficient folia (cdf) is a recessive mouse mutation causing ataxia and cerebellar cytoarchitectural abnormalities, including hypoplasia, foliation defects, and Purkinje cell ectopia. To identify the cdf gene, we have generated a high-resolution genetic map of a 3.24 +/- 0.55 cM (95% CI) region encompassing the cdf gene using 1997 F2 mice generated from a (C3H/HeSnJ-cdf/cdf x CAST/Ei)F1 intercross. Linkage analysis showed that the cdf gene cosegregates with D6Mit208, D6Mit359, and D6Mit225. A contig of five YACs, nine BACs, and three P1s was constructed across the cdf nonrecombinant region. Based on genetic and physical maps, the cdf gene was localized to the 0.28 +/- 0.23 cM (95% CI) interval between D6Mit209 and D6Ack1. These results will greatly facilitate the map-based cloning of the cdf gene, which in turn should further knowledge of the molecular mechanisms of neuronal positioning and foliation during cerebellar development. PMID- 11013082 TI - Functional characterization of the gene encoding RLIM, the corepressor of LIM homeodomain factors. AB - RLIM is a RING H2 zinc finger protein that acts as a negative coregulator for LIM homeodomain transcription factors. We have isolated genomic lambda clones that cover the entire mouse RLIM-encoding Rnf12 gene. The Rnf12 gene encompasses 20 kb and consists of at least five exons and four introns. Several transcriptional start sites within a 24-bp region were mapped around 300 nt upstream of the translational start site. Rnf12-specific mRNA can be detected in many tissues as revealed by Northern blot analysis. Transient cotransfections reveal that the proximal Rnf12 promoter can be activated in vitro by ubiquitously and more restrictively expressed transcription factors, some of which are known mediators of signal transduction pathways, e.g., mammalian Kruppel-like transcription factors, Sox and ets-related proteins, and RBP-J. We isolated a cDNA encoding human RLIM, which is highly conserved with mouse and chick RLIM. By fluorescence in situ hybridization and interspecific backcross analysis, we have localized the Rnf12 gene to the central regions of mouse and human chromosome X. PMID- 11013085 TI - BAC contig from a 3-cM region of mouse chromosome 11 surrounding Brca1. AB - Even with the completion of a draft version of the human genome sequence only a fraction of the genes identified from this sequence have known functions. Chromosomal engineering in mouse cells, in concert with gene replacement assays to prove the functional significance of a given genomic region or gene, represents a rapid and productive means for understanding the role of a given set of genes. Both techniques rely heavily on detailed maps of chromosomal regions, initially to understand the scope of the regions being modified and finally to provide the cloned resources necessary to allow both finished sequencing and large insert complementation. This report describes the creation of a BAC clone contig on mouse chromosome 11 in a region showing conservation of synteny with sequences on human chromosome 17. We have created a detailed map of an approximately 3-cM region containing at least 33 genes through the use of multiple BAC mapping strategies, including chromosome walking and multiplex oligonucleotide hybridization and gap filling. The region described is one of the targets of a large effort to create a series of mice with regional deletions on mouse chromosome 11 (33-80 cM) that can subsequently be subjected to further mutagenesis. PMID- 11013086 TI - Molecular cloning of ring finger protein 21 (RNF21)/interferon-responsive finger protein (ifp1), which possesses two RING-B box-coiled coil domains in tandem. AB - We have cloned the full length of a novel cDNA, named ring finger protein 21 (RNF21), composed of the RING finger-B box-coiled coil (RBCC) domain and the B30.2 domain, which are characteristic of the RBCC-B30.2 family. As a structural feature, the RNF21 cDNA possessed at least three kinds of isoforms, due to alternative splicing, consisting of the long form with the RBCC-RBCC-B30.2 domain, the medium form with the RBCC-B30.2 domain, and the short form with only the RBCC domain. Moreover, respective transcripts corresponding to the three isoforms were detected in various human organs by reverse transcription-PCR and Northern blot analyses. Interestingly, the medium form of the RNF21 mRNA expressed most predominantly was dramatically up-regulated within 8-16 h by interferon stimulation of HeLa cells. These findings suggest that RNF21 is a downstream gene that may mediate interferon's biological action. PMID- 11013087 TI - Curative treatment of cerebral arteriovenous malformations by embolisation using cellulose acetate polymer followed by surgical resection. AB - AIM: To investigate the usefulness of embolising cerebral arteriovenous malformations (AVMs) with a cellulose acetate polymer solution before surgical resection. METHODS: The cases of 12 patients with AVMs treated by embolisation before surgical resection were renewed. Two types of cellulose acetate polymer solutions were used to occlude 40 feeding vessels. All patients underwent surgical resection 1-51 days after embolisation. RESULTS: Reduction of the nidus volume after embolisation ranged from 20% to nearly 100%. Transient neurological deficits occurred in three patients, persistent deficits occurred in one and there were no haemorrhagic complications. All but one arteriovenous malformation were completely resected. Embolisation helped to identify feeding vessels and ease dissection. Histopathological examination of resected specimens disclosed mild inflammatory reactions in the acute stage and no unfavourable granulomatous changes in the chronic stage. CONCLUSION: Embolisation with cellulose acetate polymer solutions followed by surgical resection is safe and efficacious for treating cerebral AVMs. PMID- 11013088 TI - Seizure control after radiosurgery on cerebral arteriovenous malformations. AB - Among 462 cases of cerebral arteriovenous malformation (AVM) treated with gamma radiosurgery, the initial presentations were haemorrhage in 68%, epilepsy in 12.8%, neurological deficits in 3.2%, minor symptoms in 7.6% and asymptomatic in 4.5% respectively. There were 79 cases (17.1%) who had had a convulsive seizure before radiosurgery and they were classified into two groups: 58 cases presented with seizure as an initial symptoms (group A) and the other 21 cases mostly had seizures following intracranial haemorrhage (group B). Before radiosurgery, generalised seizure was the predominant seizure pattern in both groups, followed by pure partial and complex partial seizures. There was no major difference in seizure patterns or seizure frequency in group A and B. At radiosurgery AVMs were treated with a mean maximum dose of 37.2 Gy and a marginal dose of 19.8 Gy. Seizures had apparently decreased in most of the cases at the last follow-up (mean 24 months) according to the obliteration of the nidus. Seizures were either decreased or had disappeared in 91.6% of group A and 62.5% of group B patients. The overall results indicate that seizures improved in 85.5%, were changed in 11.6% and deteriorated in 2.9% of patients. Radiosurgery is effective not only for the obliteration of nidus of cerebral AVM, but also for seizure control, even before complete occlusion of the nidus. PMID- 11013089 TI - Image-guided microsurgery with the Mehrkoordinaten Manipulator system for cerebral arteriovenous malformations. AB - Four patients with cerebral arteriovenous malformations (AVMs) underwent image guided microsurgery with the Mehrkoordinaten Manipulator (MKM) system, which integrates a robotic microscope with a computer workstation. The patients were all male, from 8 to 51 years old (mean = 24), all presenting with intracerebral haemorrhage. The lesion was located in the deep sylvian fissure in one patient, the fronto-parieto-occipital area in one and the trigone in two. Stereotactic computed tomography and magnetic resource imaging of 1-mm slices were taken. The extent of AVM and the draining vein, predetermined with the MKM workstation, could be superimposed on the microscopic view, resulting in minimum scalp incision and craniotomy, as well as allowing for a stereotactic approach to deep seated lesions. Superimposition of the contour of the lesion was also useful for resecting the lesion, although intraoperative diagnosis of the total resection required intraoperative digital subtraction angiography. In conclusion, image guided microsurgery with the MKM system can assist minimally invasive and maximally effective microsurgery for cerebral AVMs. PMID- 11013090 TI - Endovascular treatment of aneurysms associated with cerebral arteriovenous malformations: experiences after the introduction of Guglielmi detachable coils. AB - The authors have treated 172 patients with arteriovenous malformation (AVM) since 1993. Among them, 25 patients had aneurysms with a total number of 43. The aneurysms were divided into four groups; proper feeder aneurysm (4), flow-related distal aneurysm (beyond the circle of Willis or M1, 7), flow-related proximal aneurysm (26) and remote aneurysm (6). Guglielmi detachable coil (GDC) embolisation was performed in 12 patients with 15 aneurysms. Gamma knife radiosurgery for AVM was performed in 10 of those 12 patients. Two of the proper feeder aneurysms were embolised with liquid material, together with the corresponding part of the AVM. The other aneurysms were treated surgically (14) or observed (12). There was no bleeding from aneurysms after treatment. In conclusion, GDC embolisation is a useful treatment for aneurysms associated with AVM, especially if the AVM is treated by radiosurgery. Liquid embolisation of a proper feeder aneurysm is one of the treatment options. Ltd. PMID- 11013091 TI - Surgery or gamma -knife for the treatment of arteriovenous malformations? AB - Decision making for either surgery or gamma-knife for the treatment of arteriovenous malformations (AVMs) cannot be uniform. The skill of the neurosurgeon in operating on AVMs is now being compared with that of the gamma knife. The decision varies from case to case and is to be taken by the neurosurgeon. This report presents three cases in which such decision making was not easy. Case 1 was a non-ruptured cingulate AVM of 2.5 cm diameter in the cingulate cortex. The operative field was anticipated to be very narrow between the parietal bridging veins. Case 2 was a tiny ruptured AVM in the speech-motor area which was buried underneath the cortex. Case 3 was a large ruptured thalamo stiriate-capsular AVM with feeders from the anterior and posterior choroidal arteries. All cases were operated without serious morbidity. A combination of pre operative intravascular surgery (cases 1 and 3) or postoperative gamma-knife (case 3) was adopted. In conclusion, there is no unitary rule to decide on surgery or gamma-knife for the treatment of AVMs. It depends on what good or harm the responsible surgeon or the gamma-knife does. PMID- 11013092 TI - Treatment of arteriovenous malformation of the brain--preliminary experience. AB - With the availability of new techniques, such as intravascular embolisation and radiosurgery, the therapeutic approach to arteriovenous malformations (AVMs) of the brain has recently been modified. The present study reports the authors, experiences in treating AVMs over the past 13 years. Spetzler-Martin grading of AVMs was I and II in 19 cases, III in 12, IV in 5 and V in 1 case. Four therapeutic regimens were utilised: surgical resection alone, embolisation and resection, and radiosurgery alone or after surgical resection. Generally, for low grade AVMs (Spetzler-Martin grades I, II and III), the therapeutic choice was surgical resection in 27 cases, in combination with pre-operative embolisation in two of these patients. Two cases received radiotherapy only and one case received radiosurgery after embolisation, while one case was treated conservatively. Of the five cases of grade IV, four required surgical treatment, whereas the fifth case was treated conservatively. Favourable results (good recovery and moderate disability) were obtained in 96% of the low-grade AVMs as compared with the high grade AVMs (66%) that had a poor outcome (due to primary brain damage resulting from haemorrhage at the onset in three cases and due to postoperative re-bleeding in one case). This report summarises preliminary experience in treating intracranial AVMs by surgical resection, intravascular embolisation and radiotherapy. Good therapeutic results can be expected by combining these therapeutic modalities. PMID- 11013093 TI - Successful surgical treatment of a large mixed pial-dural arteriovenous malformation. AB - A large mixed pial-dural arteriovenous malformation was successfully treated by surgical resection and occlusion of the draining veins. Treatment of this type of malformation is discussed. PMID- 11013094 TI - Surgical management of cerebral arteriovenous malformations with intraoperative digital subtraction angiography. AB - Complete excision of cerebral arteriovenous malformations (AVMs) is requisite to improving the outcome of patients with AVMs. Five patients with a small or medium cerebral AVM underwent surgery with an intraoperative digital subtraction angiography (DSA) unit. There were no residual AVMs and no complications in the examination of intraoperative DSA. The findings of postoperative angiography were consistent with those of intraoperative DSA. Intraoperative DSA provided the benefits of not only identification of a feeding artery, but also recognition of the complete excision during surgery. PMID- 11013095 TI - Dural arteriovenous malformations associated with cerebral aneurysms. AB - Only limited information is available concerning dural arteriovenous malformations (AVMs) found in association with cerebral aneurysms. The present report focuses on six such cases, concentrating attention on clinical characteristics and significance. Of a total of 46 dural AVMs encountered over a given period, six (13%) were linked with cerebral aneurysms. Particularly strong associations were noted for dural AVMs in the anterior cranial fossa (three of four cases) and convexity (all of three cases). With dural AVMs in the anterior cranial fossa, subarachnoid haemorrhage is common, so that where this is encountered the possibility of a complicating aneurysm should be considered in order to select an appropriate treatment. PMID- 11013096 TI - Haemodynamic evaluation of cerebral arteriovenous malformations by quantification of transit time using high speed digital subtraction angiography: basic considerations. AB - It is important to evaluate the haemodynamics of cerebral arteriovenous malformations (AVMs) in order to predict and avoid complications following surgical intervention. This study measured the transit time of each component of AVMs by high-speed digital subtraction angiography (DSA) to understand the haemodynamics and to evaluate which parameter is an appropriate indicator for the monitoring. Eleven cases of cerebral AVMs were studied. Haemodynamics were evaluated by analysis of tracer transit using DSA. Tracer transit parameters including transit time and peak time were calculated by application of gamma fitting to time density curve, and measured in the following compartments; feeding artery, nidus and draining vein. Significant correlations of tracer transit parameters were present among compartments. Although there was no significant correlation of nidus volume and tracer transit parameters, the peak time was shorter in haemorrhage cases than in non-haemorrhage cases. Measurements of tracer transit parameters by high-speed DSA may be a useful tool for characterising and monitoring the haemodynamics of AVMs following surgical intervention. PMID- 11013097 TI - Colour Doppler flow imaging of the superior ophthalmic vein in dural arteriovenous fistulas before and after surgery. AB - The aim of this study was to examine the clinical significance of the colour Doppler flow imaging (CDFI) findings of the superior ophthalmic vein (SOV) in intracranial dural arteriovenous fistulas (DAVF). The SOV was examined by CDFI in 12 cases of DAVF before and after surgery. Before surgery, the average SOV diameter was 3.57+/-1.18 mm (mean +/- standard deviation, which was significantly wide (P<0.05) compared with the control value. One case showed reversed flow. Four cases showed an abnormal waveform. The cases with the more severe clinical symptoms showed wider SOV diameters and more abnormal waveforms than those with mild clinical symptoms. Postoperatively, the mean SOV diameter and mean resistance index improved significantly (P< 0.05); the flow direction and waveform became normal in each. The SOV CDFI findings were found to be useful as screening and follow-up techniques for the intracranial DAVFs. PMID- 11013098 TI - Surgery for dural arteriovenous fistula in superior sagittal sinus and transverse sigmoid sinus. AB - The aim of this study was to evaluate the outcome of surgically treated dural arteriovenous fistula (DAVF). The authors performed surgical removal of DAVF in 12 patients. The locations of DAVF were the transverse sigmoid sinus in seven patients and superior sagittal sinus in five patients. These 12 patients had undergone endovascular embolisation prior to removal. Among them, six patients were completely cured, according to angiography immediately after embolisation, but these six patients showed the recurrence of DAVF within 1 year. The other six patients showed a decrease of feeding vessels. Therefore, all 12 patients underwent surgical removal of DAVF. The surgical strategies were as follows. The feeding vessels and the cortical veins with retrograde filling were occluded and cut. The affected sinus was skeletonised, and if it was occluded or almost occluded, the sinus was removed. Postoperatively, transient aphasia was seen in one patient. There was no surgical morbidity or mortality. During the follow-up period (mean 2.9 years), no recurrence of DAVF was seen. Surgical treatment is a safe and effective treatment manoeuvre for DAVF around the transverse sigmoid sinus and superior sagittal sinus. PMID- 11013099 TI - Spinal intradural perimedullary arteriovenous fistula with varix in infant. AB - A rare occurrence of type IV spinal arteriovenous malformation (intradural perimedullary arteriovenous fistula) is described in an 18-month-old boy initially misdiagnosed with Guillain-Barre syndrome. An intramedullary mixed intensity mass lesion at Th1 was demonstrated by magnetic resonance imaging together with flow voids over the dorsal aspect of the swollen spinal cord. Angiography demonstrated an intradural perimedullary arteriovenous fistula including an intraparenchymal vascular pocket. After partial embolisation of the posterior spinal arteries through the left intercostal-radicular artery, the arteriovenous fistula was removed completely together with an organised haematoma. The fistula directly opened into a vascular pocket, which was confirmed pathologically to be a varix. The postoperative course was uneventful, and the patient resumed ambulation within 4 months. The case, subclassifiable as a type IVb spinal perimedullary AVF, was unique given its location and the patient's age at presentation. PMID- 11013100 TI - Combination of intraoperative embolisation with surgical resection for treatment of giant cerebral arteriovenous malformations. AB - OBJECTIVE: To reduce the risk of surgical resection of giant arteriovenous malformation (AVM) and prevent normal perfusion pressure breakthrough (NPPB) and thus to lower postoperative mortality. METHODS: During the operation, which was carried out under general anaesthesia, the proximal ends of the feeding arteries were first ligated and 0.5 ml IBCA mixed with 0.5 ml of 5% glucose was injected into the vessels towards the AVM, then the malformed vessels were totally resected. Postoperative digital subtraction angiography (DSA) of the four vessels was performed in all patients. RESULTS: Fifty patients with giant AVMs survived after operation, only 6 (12.0%) had transient neurological dysfunction and 44 (88.0%) recovered after a follow-up of 6-36 months. No patient suffered from NPPB. CONCLUSIONS: The embolisation could block the arteriovenous shunts sufficiently to decrease the blood flow away from the normal areas of the brain so as to prevent the incidence of intra- and post-operative rebleeding, especially in NPPB. Therefore, the combination of intraoperative embolisation with surgical resection is an effective strategy in the treatment of giant cerebral AVMs, which makes it possible to operate on patients who used to be regarded as inoperable cases. PMID- 11013101 TI - Strategy for the treatment of arteriovenous malformations. AB - The treatment of arteriovenous malformations (AVMs) is still a challenging problem in the neurosurgical field. The deep-seated AVMs are a definite indication for radiosurgery for the small AVMs and with pre-embolisation for the large AVMs. The superficial AVMs are a good indication for surgery. In the case of small AVMs, surgery alone is a viable option; however, in the case of large AVMs, pre-operative embolisation is essential for prevention of NPPB (normal perfusion pressure breakthrough). Embolisation alone cannot be used, except for a small AVM in the non-eloquent cortex. Preoperative embolisation makes surgery easy; however, it causes the surrounding cortex to infarct. Hyperperfusion may occur after the direct removal of high-flow large AVMs, therefore postoperative management will be difficult in these cases. In eloquent cortex minimally invasive surgery is more reliable with respect to the morbidity produced. Therefore in cases of small AVMs in the functional cortex, direct surgery is the only choice. In cases of high-flow large AVMs, surgery and postoperative management are risky because of NPPB. Therefore pre-operative embolisation followed by surgery is a better choice. In high-flow AVMs, local blood circulation is not decreased by temporary clipping of the feeding arteries. So we recommend temporary clipping of all feeding arteries, even away from the nidus where it is easier to control bleeding. PMID- 11013102 TI - Palliatively treated cerebral arteriovenous malformations: follow-up results. AB - The purpose of this study was to evaluate the benefits of palliative embolisation of patients with inoperable intracranial AVMs. It involved the analysis of the long-term clinical follow-up results of 27 patients with inoperable intracranial AVMs. Sixteen patients were treated medically and 11 patients received partial embolisation. The mean size of the lesions was 7.2 +/- 2.6 cm, and the mean follow-up period was 99 +/- 44 months (range 52-192 months). There were no significant statistical differences in the sex, age, size, venous drainage pattern, location of the lesions or presented symptoms. The analysis of the long term clinical follow-up results showed no significant difference in either the risk of haemorrhage or clinical status of the patients in the two treatment groups. Of the 16 patients in the medical treatment group, 8 (50%) showed a clinical improvement, 3 no improvement and 5 (31%) deterioration. Of the 11 patients in the embolisation group, 5 (45.5%) showed clinical improvement, 3 no improvement and 3 (27.3%) deterioration (P = 0.871). Twenty-five percent (4 cases) of patients in the medical group and 45.5% (5 cases) of patients in the embolisation group suffered from haemorrhage during the follow-up period (P = 0.270). Complications related to embolisation occurred in three cases. This comparative study shows that palliative partial embolisation of intracranial AVMs, in all probability does not produce better clinical results than medical treatment. PMID- 11013103 TI - Haemorrhagic complication after total extirpation of huge arteriovenous malformations. AB - Two cases with huge arteriovenous malformations (AVMs) who developed haemorrhagic complications after surgery are described. The cause of the postoperative haemorrhage was considered to be the normal perfusion pressure breakthrough phenomenon and/or occlusive hyperaemia. These two haemodynamic insults possibly occur simultaneously and induce life-threatening haemorrhage. It was concluded that a huge high-flow AVM with a large venous ampulla in its deep drainers has a high risk of a postoperative intravenous thrombosis, resulting in haemorrhage together with normal perfusion pressure breakthrough phenomenon. PMID- 11013104 TI - Resectability of Spetzler-Martin grade IV and V cerebral arteriovenous malformations. AB - This study retrospectively analysed treatment modalities and outcomes in 63 cases of high grade cerebral arteriovenous malformations (Spetzler-Martin's classification grade IV 51, V 12) who were admitted to this institute between January 1986 and April 1998. Twenty-seven of 37 cases with haemorrhagic onset received surgical resection with or without pre-operative embolisation. Total extirpation was achieved in 23 cases, and small residual nidus was treated by postoperative radiosurgery in two of four subtotally resected cases. Another 6 of 37 haemorrhagic cases received radiosurgery with or without preradiosurgical embolisation. Four cases with haemorrhagic onset received partial embolisation. Finally, 29 of 37 haemorrhagic cases obtained complete disappearance of nidus. On the other hand, 4 of 26 non-haemorrhagic cases received surgical total resection, 5 radiosurgery, 9 partial embolisation and 8 observation without treatment. Microsurgical morbidity was 9% and mortality was 0%. Post-treatment haemorrhage was seen in 11 cases (1 after subtotal resection, 4 after radiosurgery, 6 after partial embolisation) and was fatal in 3 cases. Surgical resection can be achieved with acceptable morbidity, and complete obliteration of nidus should be accomplished in any treatment modality to avoid post-treatment haemorrhage. PMID- 11013105 TI - Embolisation of cerebral arteriovenous malformations to assure successful subsequent radiosurgery. AB - This study investigated the angiographic changes in embolised arteriovenous malformations (AVMs) pre- and post-embolisation and preradiosurgery to clarify the usefulness of embolisation as a pretreatment for radiosurgery and the strategy of embolisation for the radiosurgical success. A total of 37 patients with cerebral AVMs treated over a period of 4 years was investigated. All the AVMs were embolised with N-butyl cyanoacrylate and 2 months later they were treated by radiosurgery. The size of AVM nidus reduced just following the embolisation (mean 21.9 ml to 3.9 ml). The angiogram taken in preparation for radiosurgery showed a further size reduction in the nidus of 16 AVMs, no change in 10 and regrowth in 11. In all the cases where size was reduced, the nidus was densely packed, while all the regrown AVMs were of the diffuse type. Five AVMs disappeared following radiosurgery, all of which were size-reduction or no-change cases. In conclusion, to achieve success in subsequent radiosurgery, nidus embolisation and the occlusion of fistulous and meningeal feeders are mandatory. Imprudent proximal feeder occlusion and the use of embolic materials with a risk of recanalisation should be avoided to prevent regrowth of the nidus, which may lead to errors in planning the radiosurgery to follow. PMID- 11013106 TI - Strategic embolisation for successful resection of a large cerebral arteriovenous malformation. AB - The risks accompanied by the treatment of cerebral arteriovenous malformation (AVM) are still cumulative despite recent progress in available treatment options. Pre-operative embolisation is one such option, however, it seldom makes the surgical resection difficult. The excessive embolised nidus makes the surgical resection difficult because it cannot be compressed during the resection surgery and embolised nidus as a 'glue ball' with marginal hypervascular territory is most difficult to remove. The aim of pre-operative embolisation for successful surgical resection is to put glue into the marginal part of the nidus so as to make a cleavage between the surrounding normal tissues. Remaining feedings via the dilatated leptomeningeal anastomoses from surrounding normal cortical arteries do not interfere with the resection and can be eliminated easily by coagulating the pia matter around the nidus. Strategic planning with regard to the systemic course of treatment, including the manner of resection, is important for effective pre-operative embolisation. PMID- 11013107 TI - Surgical resection of cerebral arteriovenous malformation combined with pre operative embolisation. AB - To assess the importance of pre-operative embolisation, 27 cases of cerebral artriovenous malformation (AVM) treated in this institute between July 1994 and October 1998 were analysed. The patients' ages ranged from 3 to 70 years (average 36.9) with a follow-up period of 1-41 months (average 19.2). The patient presented with haemorrhage in 21 cases and seizure in five. In 21 of 27 cases, surgical resection of a nidus was performed, gamma knife therapy was applied in three and conservative therapy was chosen in three. Of 21 cases treated surgically, total removal was achieved in 19 cases and a residual nidus was seen in one (a large basal ganglia AVM). In the remaining case, postoperative angiography was not available. Pre-operative embolisation followed by surgical resection of the nidus was performed in seven cases in which there was a large AVM. A volume index was calculated to indicate the size of the nidus using X x Y x Z, where X is the maximum diameter (cm) of the nidus on the lateral angiogram, Y is the diameter (cm) perpendicular to X and Z is the maximum diameter (cm) on the anteroposter or angiogram. The index averaged 45.9 for the cases in which pre operative embolisation was performed, while it was 5.6 in the cases without embolisation. Pre-operative embolisation was performed to reduce the nidus flow as much as possible, to prevent overload to the surrounding structures. At surgery, the nidus was resected from the surrounding tissue and care was taken not to enter the nidus. Postoperatively, the systolic blood pressure was maintained at 90-100 mmHg for several days in the intensive care unit. The results were excellent in 15 cases, good in three (hemiparesis due to the initial haemorrhage remained in all three), fair in one (a patient with a severe subarachnoid haemorrhage). Two patients died (acute pulmonary oedema and severe meningitis). Minor postoperative bleeding or oozing was seen in three cases. In conclusion, reducing the shunt flow through a nidus in a step-wise fashion with pre-operative embolisation of a large AVM seems to be quite helpful in preventing postoperative haemodynamic overload to the surrounding brain. It is also important not to enter the nidus when it is removed at surgery. This helps to prevent intraoperative and/or postoperative bleeding, and led to successful total removal of the nidus with a good postoperative course. PMID- 11013108 TI - Defective deletion mutant amplification. AB - Defective deletion mutants can be replicated in superinfected cells by parasitism of the intact virus' replication machinery, and through replication with the host cell. We show by analysis of a mathematical model that dynamic stability of superinfected cell growth is crucial in determining the frequency of deletion mutant infected cells, i.e. there is a critical infectivity threshold rho(sc)below which the density of proliferative virus is significantly reduced by the presence of defective deletion mutants. Above rho(sc), proliferative virus principally occurs as superinfected cells (wild type with defective deletion mutant). The threshold rho(sc), and the interference effects of the deletion mutant, increase with deletion mutant parasitism of the wild-type replication machinery in superinfected cells. The interaction of virally infected cells with host homeostasis determines whether immune escape by deletion mutant infected cells is necessary for the interference window to exist. Only when the deletion mutant has a detrimental effect on infected host cell replication did we observe periodic behaviour. PMID- 11013109 TI - On the dynamic persistence of cooperation: how lower individual fitness induces higher survivability. AB - We study a model in which cooperation and defection coexist in a dynamical steady state. In our model, subpopulations of cooperators and defectors inhabit sites on a lattice. The interactions among the individuals at a site, in the form of a prisoner's dilemma (PD) game, determine their fitnesses. The chosen PD payoff allows cooperators, but not defectors, to maintain a homogeneous population. Individuals mutate between types and migrate to neighboring sites with low probabilities. We consider both density-dependent and density-independent versions of the model. The persistence of cooperation in this model can be explained in terms of the life cycle of a population at a site. This life cycle starts when one cooperator establishes a population. Then defectors invade and eventually take over, resulting finally in the death of the population. During this life cycle, single cooperators migrate to empty neighboring sites to found new cooperator populations. The system can reach a steady state where cooperation prevails if the global "birth" rate of populations is equal to their global "death" rate. The dynamic persistence of cooperation ranges over a large section of the model's parameter space. We compare these dynamics to those from other models for the persistence of altruism and to predator-prey models. PMID- 11013110 TI - Sperm competition games: a comparison of loaded raffle models and their biological implications. AB - Our main aim is to compare the additive model, due to Mesterton-Gibbons, and the multiplicative model, due to Parker, of sperm allocation under sperm competition, when other influences are treated in the same way. We first review these (and other) models and their foundations, leading to a generalization of the multiplicative model. Sperm is assumed to cost energy, and this constraint is incorporated differently in the two models. These give the same results in the random-roles situation when the males occupy roles (of first and second to mate) randomly: the number of sperm ejaculated in the favoured role is greater than that in the disfavoured role by an amount that depends on the effect of sperm limitation (i.e. the probability that there is insufficient sperm to ensure full fertility). If the latter is negligible, or the fertilization raffle fair, this difference is zero, as Parker found originally. In the constant roles situation (where males of a particular type always occupy the same role) the predictions differ: the additive model has the same predictions as in the random roles case, but the multiplicative model predicts that males of the type occupying the favoured role ejaculate less than males of the type occupying the disfavoured role, in accord with Parker's original conclusion. The fitnesses of the two types of male can be calculated in the multiplicative model: the fitness of the favoured male is usually higher, even if he has to expend more energy in "finding" a female, e.g. through fighting, etc. These conclusions relate to inter male behaviour (i.e. of different male types), as distinct from intra-male behaviour (i.e. of a given male when in different roles). We analyse situations in which one male type has some probability of acting in its less usual role: calculations with varying amounts of sperm limitation are presented. It is found that the presence of a male of a different type has an effect on intra-male ejaculate behaviour, which also depends critically on the role usually occupied. We conclude that the multiplicative model is the more accurate model and provides more information. Some experimental data on sperm numbers are used to find the effects of sperm limitation. For species which conform to the loaded raffle model, sperm limitation typically has small or negligible effects: in this case, we argue that empiricists should look for equal ejaculates in the two roles when studying random role situations; when roles are occupied non-randomly average sperm expenditure should be greater by male types typically occupying the disfavoured role, but within a male type, expenditure should be greater in the role it typically occupies. PMID- 11013111 TI - A mathematical model describing kinetics of conversion of violaxanthin to zeaxanthin via intermediate antheraxanthin by the xanthophyll cycle enzyme violaxanthin de-epoxidase. AB - The xanthophyll cycle is one of the mechanisms protecting the photosynthetic apparatus against the light energy excess. Its action is still not well understood on the molecular level. Our model makes it possible to follow independently the kinetics of the two de-epoxidation steps occurring in the xanthophyll cycle: the conversion of violaxanthin into antheraxanthin and the conversion of antheraxanthin into zeaxanthin. Using a simple form of the transition rates of these two conversions, we model the time evolution of the concentration pattern of violaxanthin, antheraxanthin and zeaxanthin during the de-epoxidation process. The model has been applied to describe the reactions of de-epoxidation in a system of liposome membranes composed of phosphatidylcholine and monogalactosyldiacylglycerol. Results obtained within the model fit very well with the experimental data. Values of the transition probabilities of the violaxanthin conversion into antheraxanthin and the antheraxanthin conversion into zeaxanthin calculated by means of the model indicate that the first stage of the de-epoxidation process is much slower than the second one. PMID- 11013112 TI - Coupled biological oscillators in a cave insect. AB - Insects that live in the interior of caves show the basic internal temporal organization of coupled oscillators. An analysis is made of the coupled moulting and oviposition cycles of Folsomia candida, a cave-dwelling Collembolan, with regard to their oscillatory nature, their phase dependent responses to external perturbations, the effect of coupling on these responses, and conjecture about the link of these cycles with circadian clocks in other organisms. PMID- 11013113 TI - Optimization of coding potentials using positional dependence of nucleotide frequencies. AB - We study the coding potential of human DNA sequences, using the positional asymmetry function (D(p)) and the positional information function (I(q)). Both D(p)and I(q)are based on the positional dependence of single nucleotide frequencies. We investigate the accuracy of D(p)and I(q)in distinguishing coding and non-coding DNA as a function of the parameters p and q, respectively, and explore at which parameters p(opt)and q(opt)both D(p)and I(q)distinguish coding and non-coding DNA most accurately. We compare our findings with classically used parameter values and find that optimized coding potentials yield comparable accuracies as classical frame-independent coding potentials trained on prior data. We find that p(opt)and q(opt)vary only slightly with the sequence length. PMID- 11013114 TI - Cytokine-modulated regulation of helper T cell populations. AB - Helper T (Th) cells are a crucial component of the adaptive immune system and are of fundamental importance in orchestrating the appropriate response to pathogenic challenge. They fall into two broad categories defined by the cytokines each produces. Th1 cells produce interferon- gamma and are required for effective immunity to intracellular bacteria, viruses and protozoa whereas Th2 produce IL-4 and are required for optimal antibody production to T-dependent antigens. A great deal of experimental data on the regulation of Th1 and Th2 differentiation have been obtained but many essential features of this complex system are still not understood. Here we present a mathematical model of Th1/Th2 differentiation and cross regulation. We model Fas-mediated activation-induced cell death (AICD) as this process has been identified as an important mechanism for limiting clonal expansion and resolving T cell responses. We conclude that Th2 susceptibility to AICD is important for stabilizing the two polarized arms of the T helper response, and that cell-cell killing, not suicide, is the dominant mechanism for Fas-mediated death of Th1 effectors. We find that the combination of the anti proliferative effect of the cytokine TGF- beta and the inhibiting influence of IL 10 on T cell activation are crucial controls for Th2 populations. We see that the strengths of the activation signals for each T helper cell subset, which are dependent on the antigen dose, co-stimulatory signals and the cytokine environment, critically determine the dominant helper subset. Switches from Th1- to Th2-dominance may be important in chronic infection and we show that this phenomenon can arise from differential AICD susceptibility of T helper subsets, and asymmetries in the nature of the cross-suppressive cytokine interactions. Our model suggests that in some senses a predominantly type 2 reaction may well be the "default" pathway for an antigen-specific immune response, due to these asymmetries. PMID- 11013115 TI - Microvillar ion channels: cytoskeletal modulation of ion fluxes. AB - The recently presented theory of microvillar Ca(2+)signaling [Lange, K. (1999) J. Cell. Physiol.180, 19-35], combined with Manning's theory of "condensed counterions" in linear polyelectrolytes [Manning, G. S. (1969). J. Chem. Phys.51, 924-931] and the finding of cable-like ion conductance in actin filaments [Lin, E. C. & Cantiello, H. F. (1993). Biophys. J.65, 1371-1378], allows a systematic interpretation of the role of the actin cytoskeleton in ion channel regulation. Ion conduction through actin filament bundles of microvilli exhibits unique nonlinear transmission properties some of which closely resemble that of electronic semiconductors: (1) bundles of microfilaments display significant resistance to cation conduction and (2) this resistance is decreased by supply of additional energy either as thermal, mechanical or electromagnetic field energy. Other transmission properties, however, are unique for ionic conduction in polyelectrolytes. (1) Current pulses injected into the filaments were transformed into oscillating currents or even into several discrete charge pulses closely resembling that of single-channel recordings. Discontinuous transmission is due to the existence of counterion clouds along the fixed anionic charge centers of the polymer, each acting as an "ionic capacitor". (2) The conductivity of linear polyelectrolytes strongly decreases with the charge number of the counterions; thus, Ca(2+)and Mg(2+)are effective modulator of charge transfer through linear polyelectrolytes. Field-dependent formation of divalent cation plugs on either side of the microvillar conduction line may generate the characteristic gating behavior of cation channels. (3) Mechanical movement of actin filament bundles, e.g. bending of hair cell microvilli, generates charge translocations along the filament structure (mechano-electrical coupling). (4) Energy of external fields, by inducing molecular dipoles within the polyelectrolyte matrix, can be transformed into mechanical movement of the system (electro-mechanical coupling). Because ionic transmission through linear polyelectrolytes is very slow compared with electronic conduction, only low-frequency electromagnetic fields can interact with the condensed counterion systems of linear polyelectrolytes. The delineated characteristics of microvillar ion conduction are strongly supported by the phenomenon of electro-mechanical coupling (reverse transduction) in microvilli of the audioreceptor (hair) cells and the recently reported dynamics of Ca(2+)signaling in microvilli of audio- and photoreceptor cells. Due to the cell-specific expression of different types and combinations of ion channels and transporters in the microvillar tip membrane of differentiated cells, the functional properties of this cell surface organelle are highly variable serving a multitude of different cellular functions including receptor-mediated effects such as Ca(2+)signaling, regulation of glucose and amino acid transport, as well as modulation of membrane potential. Even mechanical channel activation involved in cell volume regulation can be deduced from the systematic properties of the microvillar channel concept. In addition, the specific ion conduction properties of microfilaments combined with their proposed role in Ca(2+)signaling make microvilli the most likely cellular site for the interaction with external electric and magnetic fields. PMID- 11013116 TI - Regulation of platelet production: the normal response to perturbation and cyclical platelet disease. AB - An age-structured model for the regulation of platelet production is developed, and compared with both normal and pathological platelet production. We consider the role of thrombopoietin (TPO) in this process, how TPO affects the transition between megakaryocytes of various ploidy classes, and their individual contributions to platelet production. After the estimation of the relevant parameters of the model from both in vivo and in vitro data, we use the model to numerically reproduce the normal human response to a bolus injection of TPO. We further show that our model reproduces the dynamic characteristics of autoimmune cyclical thromobocytopenia if the rate of platelet destruction in the circulation is elevated to more than twice the normal value. PMID- 11013117 TI - A model of beta-cell mass, insulin, and glucose kinetics: pathways to diabetes. AB - Diabetes is a disease of the glucose regulatory system that is associated with increased morbidity and early mortality. The primary variables of this system are beta-cell mass, plasma insulin concentrations, and plasma glucose concentrations. Existing mathematical models of glucose regulation incorporate only glucose and/or insulin dynamics. Here we develop a novel model of beta -cell mass, insulin, and glucose dynamics, which consists of a system of three nonlinear ordinary differential equations, where glucose and insulin dynamics are fast relative to beta-cell mass dynamics. For normal parameter values, the model has two stable fixed points (representing physiological and pathological steady states), separated on a slow manifold by a saddle point. Mild hyperglycemia leads to the growth of the beta -cell mass (negative feedback) while extreme hyperglycemia leads to the reduction of the beta-cell mass (positive feedback). The model predicts that there are three pathways in prolonged hyperglycemia: (1) the physiological fixed point can be shifted to a hyperglycemic level (regulated hyperglycemia), (2) the physiological and saddle points can be eliminated (bifurcation), and (3) progressive defects in glucose and/or insulin dynamics can drive glucose levels up at a rate faster than the adaptation of the beta -cell mass which can drive glucose levels down (dynamical hyperglycemia). PMID- 11013119 TI - Editorial PMID- 11013120 TI - Pharmacologic relevance of K(+)Channel remodeling in atrial fibrillation. PMID- 11013121 TI - Angiotensin II induces nuclear factor- kappa B activation in cultured neonatal rat cardiomyocytes through protein kinase C signaling pathway. AB - Rat neonatal ventricular cardiomyocytes (RNVM) possess G protein-coupled AT(1)receptors for angiotensin II (AngII) that activate multiple intracellular pathways. To elucidate potential signaling mechanisms involved, we focussed on the nuclear transcription factor-kappa B (NF- kappa B) in RNVM culture. Using specific antibody to NF- kappa Bp65, immunolocalization of NF- kappa B was cytoplasmic in unstimulated cardiomyocytes, whereas NF- kappa B was translocated into the RNVM nucleus in response to AngII. This translocation was inhibited in the presence of calphostin C, a specific inhibitor of protein kinase C (PKC). Western blot analysis showed an increase of NF- kappa B in AngII-stimulated cardiomyocyte nuclear extracts as compared to controls. Biomolecular interaction analysis (BIA analysis) of NF- kappa B activation showed that only AngII-nuclear extracts bound to NF- kappa B consensus sequence with a high degree of affinity. This DNA-binding capacity was completely lost in calphostin C-treated cells. At transcriptional level in RNVM, AngII mediates the upregulation of matrix gelatinase (MMP-9), which is totally inhibited by calphostin C treatment. In conclusion, cardiomyocyte nuclear NF- kappa B translocation in response to Ang II via PKC pathway activates cardiomyocyte-specific transcription of MMP-9 and may activate transcription from responsive genes which are involved in cardiac hypertrophy process and/or cardiac remodeling. PMID- 11013122 TI - Neonatal mouse cardiac myocytes exhibit cardioprotection induced by hypoxic and pharmacologic preconditioning and by transgenic overexpression of human Cu/Zn superoxide dismutase. AB - Although mouse models have been increasingly used for studies of cardiac pathophysiology, there is little information regarding cultured murine cardiac myocytes. Accordingly, we have developed a cell culture model of neonatal mouse cardiac myocytes by modifying a protocol used to prepare neonatal rat myocytes. The principal change is the substitution of cytosine arabinoside for bromodeoxyuridine to prevent fibroblast proliferation. Neonatal murine myocytes exhibited persistent spontaneous contraction and were viable for up to 14 days in culture. By flow cytometry 85% of the cells were cardiac myocytes. In sparse cultures (average cell density 259 cells/mm(2)), both hypoxic preconditioning (n=5) and phenylephrine pretreatment (n=8) produced significant protection of cardiac myocytes from cell death during a prolonged period of severe hypoxia (<0.5% O(2)for 18-20 h, both P<0.05). The phenylephrine effect was inhibited by the alpha(1)-adrenoceptor antagonist prazosin (n=4, P<0.05) and by an xi PKC peptide antagonist (xi V1-2) coupled to a TAT peptide (n=5, P<0. 05). Interestingly, the mixed alpha(1)- and beta -adrenoceptor agonist norepinephrine, which stimulates hypertrophy as measured by(14)[C]phenylalanine incorporation in neonatal rat cardiac myocytes, did not cause hypertrophy in mouse myocytes, suggesting that the signaling pathways for myocardial protection and hypertrophy are likely to be both divergent and species specific. In cardiac myocytes prepared from transgenic mice either homozygous or heterozygous for human Cu/Zn superoxide dismutase, there was protection from cell death (n=3) and restoration of(14)[C]phenyl- alanine uptake (n=4) during prolonged hypoxia (1% O(2)for 3 days, both P<0.05). We conclude that this cellular model, which is relatively simple to prepare, can be used for in-vitro examination of cardiac protection induced by preconditioning agents, various transgenes, and potentially by targeted gene deletions. PMID- 11013123 TI - The stress-responsive MAP kinase p38 is activated by low-flow ischemia in the in situ porcine heart. AB - Stress-responsive p38 MAP kinase is activated by phosphorylation during global and severe regional myocardial ischemia. However, it is unknown whether or not moderate, low-flow ischemia also activates p38 MAP kinase. Therefore, we investigated p38 MAP kinase activation in an established model of short-term hibernation and stunning. In anesthetized swine, coronary blood flow into the left anterior descending coronary artery was decreased in order to reduce regional contractile function by identical with 50%. Transmural myocardial biopsies were taken before (controls) and during ischemia as well as after reperfusion. Creatine phosphate content, after an early ischemic reduction, recovered to control values at 90 min ischemia. The expression of phospholamban, SERCA2a, calsequestrin, and troponin inhibitor was unchanged under these conditions (Northern and Western blotting). At 8 min of ischemia, however, p38 MAP kinase was activated to 221% of the pre-ischemic value as judged by its elevated phosphorylation state. Then, it returned to control values by 85 min ischemia. We conclude that low-flow ischemia transiently activates the stress responsive p38 MAP kinase which might act to trigger cardioprotective events. PMID- 11013124 TI - Pre-synaptic NO-cGMP pathway modulates vagal control of heart rate in isolated adult guinea pig atria. AB - The role of nitric oxide (NO) in the vagal modulation of heart rate (HR) is controversial. We tested the hypothesis that NO acts via a pre-synaptic, guanylyl cyclase (GC) dependent pathway. The effects of inhibiting NO synthase (NOS) and GC were evaluated in isolated atrial/right vagal nerve preparations from adult (550-750 g) and young (150-250 g) female guinea pigs. Levels of NOS protein were quantified in right atria using Western blotting and densitometry. The non specific NOS inhibitor N- omega -nitro- L -arginine (L -NA, 100 microM, n=5) significantly reduced the negative chronotropic response to vagal nerve stimulation (VNS) at 3 and 5 Hz in the adult guinea pig. This effect was reversed with 1 m ML -arginine. Similar results were observed with the specific neuronal NOS inhibitor vinyl-N5-(1-imino-3-butenyl)- L -ornithine (L -VNIO, 100 microM, n=7). Inhibition of GC with 1H-(1,2,4)-oxadiazolo-(4, 3-a)-quinoxalin-1-one (ODQ, 10 microM, n=7) also significantly reduced the negative chronotropic response to VNS at 3 and 5 Hz in adult guinea pigs. Neither L -NA (n=6), L -VNIO (n=5) nor ODQ (n=6) changed the HR response to cumulative doses of carbamylcholine in adult guinea pig atria suggesting that the action of NO is pre-synaptic. The HR response to VNS was unaffected by L -NA (n=7) or ODQ (n=7) in young guinea pigs and Western blot analysis showed significantly lower levels of nNOS protein in right atria from young animals. These results suggest a pre-synaptic NO-cGMP pathway modulates cardiac cholinergic transmission, although this may depend on the developmental stage of the guinea pig. PMID- 11013125 TI - CTGF expression is induced by TGF- beta in cardiac fibroblasts and cardiac myocytes: a potential role in heart fibrosis. AB - Connective tissue growth factor (CTGF) is a cysteine-rich protein induced by transforming growth factor beta (TGF- beta) in connective tissue cells. CTGF can trigger many of the cellular processes underlying fibrosis, such as cell proliferation, adhesion, migration and the synthesis of extracellular matrix; however, its role in acute and chronic cardiac injury is not fully understood. Here, we show that TGF- beta is a specific inducer of CTGF expression in both cardiac fibroblasts and cardiac myocytes. The activity of a CTGF promoter-based reporter construct correlated with endogenous CTGF expression, suggesting that TGF- beta induces CTGF expression most likely by activating its promoter. Upregulation of CTGF coincided with an increase in fibronectin, collagen type I and plasminogen activator inhibitor-1 production. Forskolin, a stimulator of cyclic AMP, blocked TGF- beta induced CTGF expression and reduced the basal level of CTGF, whereas an inhibitor that blocks the MAP kinase signaling pathway (PD 98059) significantly enhanced TGF- beta induced CTGF expression. Furthermore, we found that both TGF- beta and CTGF mRNAs were significantly elevated in the left ventricles and septa of rat hearts 2-16 weeks following myocardial infarction. This correlated well with concomitant increases in fibronectin, and type I and type III collagen mRNA levels in these animal hearts. Significant upregulation of CTGF was also detected in human heart samples derived from patients diagnosed with cardiac ischemia. Based on these findings, we propose that CTGF is an important mediator of TGF- beta signaling in the heart and abnormal expression of this gene could be used as a diagnostic marker for cardiac fibrosis. PMID- 11013126 TI - Augmented expression of cardiotrophin-1 and its receptor component, gp130, in both left and right ventricles after myocardial infarction in the rat. AB - Cardiotrophin-1 (CT-1) is a potent cytokine that stimulates the assembly of sarcomeric units in series in cardiomyocytes through gp130 signaling, resulting in myocardial cell hypertrophy. To clarify the role of CT-1 and the gp130 signaling pathway during ventricular remodeling after myocardial infarction, we examined the expression of CT-1 and gp130 in a rat model of myocardial infarction. At 1, 3, 7, 14, 28 and 56 days (n=12 for each group) after ligation of a coronary artery, tissue samples were obtained from infarct tissue, the ventricular septum and the right ventricle. All animals developed large myocardial infarctions, with infarct sizes ranging from 39.8% to 50.3%. Progressive left ventricular dilatation and inadequate hypertrophy of the surviving myocardium were confirmed by echocardiography. CT-1 and gp130 mRNA levels were determined by semiquantitative reverse transcription-polymerase chain reaction using 1 or 5 microg of total RNA followed by Southern blotting. The densitometric analysis of the Southern blots revealed a significant increase in CT-1 and gp130 mRNA levels (P<0.01) compared with those of the sham-operated rats at 1, 3, 7, 14, 28 and 56 days post-infarct in the infarct area, the ventricular septum (non-infarcted area) and right ventricle. The protein levels of CT-1 and gp130, determined by Western blot analysis, were significantly increased (P<0.05) compared with those of sham-operated rats, peaked during the acute stage and declined thereafter in the three regions described above. Immunohistochemical staining showed that CT-1 and gp130-immunoreactivities were detected in cardiomyocytes and fibroblast-like cells and that the intensity of staining was increased at 7 days post-infarct compared with that in sham-operated rats. An augmented CT-1 and gp130 system thus appears to play an important role during ventricular remodeling after myocardial infarction. PMID- 11013127 TI - Role of cGMP-inhibited phosphodiesterase and sarcoplasmic calcium in mediating the increase in basal heart rate with nitric oxide donors. AB - Nitric oxide (NO) donors increase heart rate (HR) through a guanylyl cyclase dependent stimulation of the pacemaker current I(f), without affecting basal I(Ca L). The activity of I(f)is known to be enhanced by cyclic nucleotides and by an increase in cytosolic Ca(2+). We examined the role of cGMP-dependent signaling pathways and intracellular Ca(2+)stores in mediating the positive chronotropic effect of NO donors. In isolated guinea pig atria, the increase in HR in response to 1-100 micromol/l 3-morpholino-sydnonimine (SIN-1; with superoxide dismutase, n=6) or diethylamine-NO (DEA-NO, n=8) was significantly attenuated by blockers of the cGMP-inhibited phosphodiesterase (PDE3; trequinsin, milrinone or Ro-13-6438, n=22). In addition, the rate response to DEA-NO or sodium nitroprusside (SNP) was significantly reduced following inhibition of PKA (KT5720 or H-89, n=15) but not PKG (KT5728 or Rp-8-pCPT-cGMPs, n=16). Suppression of sarcoplasmic (SR) Ca(2+)release by pretreatment of isolated atria with ryanodine or cyclopiazonic acid (2 micromol/l and 60 micromol/l, n=16) significantly reduced the chronotropic response to 1-100 micromol/l SIN-1 or DEA-NO. Moreover, in isolated guinea pig sinoatrial node cells 5 micromol/l SNP significantly increased diastolic and peak Ca(2+)fluorescence (+13+/-1% and +28+/-1%, n=6, P<0.05). Our findings are consistent with a functionally significant role of cAMP/PKA signaling (via cGMP inhibition of PDE3) and SR Ca(2+)in mediating the positive chronotropic effect of NO donors. PMID- 11013128 TI - Transcriptional regulation of L-type calcium channel expression in cardiac myocytes. AB - The L-type calcium channel is a heteromultimeric protein complex, which is expressed in the cardiac sarcolemma. Although post-translational regulation of its subunits by protein kinase A (PKA) has been widely reported, little is known about molecular processes that regulate expression of calcium channel subunits (alpha(1C), alpha(2)- delta, and beta(2A)subunits). Previous studies from our group demonstrate that the steady-state mRNA level of the alpha(1C)unit is increased by treatment of myocytes with beta -adrenergic agonists. The current study is designed to determine whether the mRNA levels for all subunits of the L type calcium channel are coordinately controlled by a beta -adrenergic agonist, and whether this occurs predominantly through control of rate of transcription. Nuclear run-on assays were used to determine the transcription initiation rate of these genes in cultured neonatal rat cardiac myocytes. In isoproterenol (10(-7)m) treated myocytes, transcription of genes encoding the alpha(1C), alpha(2)- delta, and beta(2A)subunits was enhanced. The increases in transcription initiation rate for alpha(1C), alpha(2)- delta, and beta(2A)subunits genes were 404%, 367%, and 240% of control, respectively. Pretreatment with the beta -adrenergic antagonist propranolol (10(-5)m) or PKA inhibitor H-89 (10(-6)m) blocked the effects of isoproterenol, while either drug alone did not affect the gene transcription rate significantly. Steady state mRNA levels of the subunits increased following isoproterenol treatment. These results suggest that beta -adrenergic stimulation and the PKA signaling pathway play an important role in transcriptional regulation of the L-type calcium channel in myocyte. The expression of all the subunits of this ion channel is under coordinate transcriptional control. PMID- 11013129 TI - Activation of the Na(+)/H(+) exchanger is required for reperfusion-induced Ins(1,4,5)P(3) generation. AB - Post-ischemic reperfusion causes a change in inositol phosphate responses to norepinephrine from primary generation of inositol(1,4) bis phosphate (Ins(1,4)P(2)) to generation of inositol(1,4,5) tris phosphate (Ins(1,4,5)P(3)) that is required for the initiation of reperfusion arrhythmias. The current study was undertaken to investigate the role of Na(+)/H(+)exchange in facilitating this transient change in inositol phosphate response. Rat hearts were subjected to 20 min ischemia followed by 2 min reperfusion and Ins(1, 4,5)P(3)content was measured by mass analysis or by anion-exchange HPLC following [(3)H]inositol labeling. Reperfusion caused generation of [(3)H]Ins(1,4,5)P(3)(1732+/-398 to 3103+/-214, cpm/g tissue, mean+/-S.E.M., n=5, P<0.01) and the development of arrhythmias. Inhibition of Na(+)/H(+)exchange, by reperfusing at pH 6.3 or by pretreating with HOE-694 (10 n M-3 microM) or HOE-642 (3 microM) prevented the [(3)H]Ins(1,4,5)P(3)generation, without causing any suppression of norepinephrine release. Increases in Ins(1,4,5)P(3)mass were similarly reduced by inhibition of Na(+)/H(+)exchange. Thus, activation of Na(+)/H(+)exchange is required for the enhanced Ins(1,4,5)P(3)response observed under reperfusion conditions, and prevention of Ins(1,4,5)P(3)generation may be an important contributor to the anti-arrhythmic actions of inhibitors of Na(+)/H(+)exchange. PMID- 11013130 TI - Metabolic inhibition activates a non-selective current through connexin hemichannels in isolated ventricular myocytes. AB - Intracellular Na(+)accumulation and K(+)loss play important roles in the pathogenesis of arrhythmias and injury in the ischemic heart. We investigated the role of metabolically sensitive connexin hemichannels as a potential route for Na(+)influx and K(+)efflux during ischemia, using dye uptake and electrophysiological measurements to assay hemichannel activity in isolated rabbit ventricular myocytes. Consistent with the known size selectivity of connexin hemichannels,;50% of myocytes exposed to either low extracellular Ca(2+)(an established method for opening connexin hemichannels) or to metabolic inhibitors (a recently described method for opening hemichannels) accumulated fluorescent dyes with <1000 MW (propidium iodide and calcein), but excluded a larger dye with 1500-3000 MW (dextran-rhodamine). Using the whole cell patch clamp technique, we found that metabolic inhibitors activated a non-selective current permeant to both small and large cations, and blocked by La(3+), similar to the properties of connexin 43 when overexpressed in human embryonic kidney (HEK) cells. These findings indicate that isolated cardiac myocytes endogenously express metabolically-sensitive connexin hemichannels. If activated during ischemia, these hemichannels could contribute significantly to altered ionic fluxes promoting arrhythmias and myocardial injury. PMID- 11013131 TI - Functional suppression of sodium channels by beta(1)-subunits as a molecular mechanism of idiopathic ventricular fibrillation. AB - Ventricular fibrillation leading to sudden cardiac death can occur even in the absence of structural heart disease. One form of this so-called idiopathic ventricular fibrillation (IVF) is characterized by ST segment elevation (STE) in the electrocardiogram. Recently we found that IVF with STE is linked to mutations of SCN5A, the gene encoding the cardiac sodium channel alpha -subunit. Two types of defects were identified: loss-of-function mutations that severely truncate channel proteins and missense mutations (e.g. a double mutation, R1232W and T1620M) that cause only minor changes in channel gating. Here we show that co expression of the R1232W+T1620M missense mutant alpha -subunits in a mammalian cell line stably transfected with human sodium channel beta(1)-subunits results in a phenotype similar to that of the truncation mutants. In the presence of beta(1)subunits the expression of both ionic currents and alpha -subunit specific, immunoreactive protein was markedly suppressed after transfection of mutant, but not wild-type alpha -subunits when cells were incubated at physiological temperature. Expression was partially restored by incubation at reduced temperatures. Our results reconcile two classes of IVF mutations and support the notion that a reduction in the amplitude of voltage-gated sodium conductance is the primary cause of IVF. PMID- 11013132 TI - The ultrarapid and the transient outward K(+) current in human atrial fibrillation. Their possible role in postoperative atrial fibrillation. AB - Atrial fibrillation (AF) causes distinct changes in atrial conduction, characterized as electrical remodeling. Experimental data on the possible significance of alterations of specific K(+)outward currents in this process are still limited in human AF. The ultra-rapid delayed rectifier current (I(Kur)) has not been studied in AF with respect to its sensitivity to 4-Aminopyridine (4-AP). To clarify the role of (1) the 4-AP sensitive I(Kur)current, compared to recordings without using 4-AP (I(Kur*)), and (2) the transient outward current (I(to)) in changes of atrial repolarization associated with AF, whole cell voltage-clamp recordings were obtained from atrial myocytes of patients undergoing elective cardiac surgery, with and without a history of atrial fibrillation (AF/non-AF). Further, a possible relation between experimental data and postoperative AF was studied. In AF patients, I(Kur*)was reduced by 40% [5.00+/-0.32 pA/pF (non-AF) and 2.91+/-0. 45 pA/pF (AF) at +50 mV, P<0.0001, n=22/11], I(Kur)by 55% [3.81+/-0. 30 pA/pF (non-AF) and 1.71+/-0.20 pA/pF (AF) at +50 mV, P<0.0001, n=22/11]. The mean amplitude of I(Kur)was significantly smaller than I(Kur*). Consistently, I(to)was reduced by 44% [11.57+/-0.77 pA/pF (non-AF) and 6.51+/-1.31 pA/pF (AF), P<0.01, n=25/11]. In 48% of non-AF patients, postoperative AF was detected. The corresponding voltage-clamp recordings showed a trend to reduced I(Kur*)and I(Kur)currents, although it did not reach statistical significance. The consistent reduction of all three K(+)currents investigated due to the presence of AF indicates an important association of abnormalities in cellular repolarization with the onset and the self-sustaining nature of human AF. PMID- 11013133 TI - Inhibition of Na(+)/H(+) exchange at the beginning of reperfusion is cardioprotective in isolated, beating adult cardiomyocytes. AB - Stimulation of Na(+)/H(+)exchange during ischemia-reperfusion results in cardiac damage. However, it is unclear whether the Na(+)/H(+)exchanger is active during the ischemic period or during reperfusion. Adult beating cardiomyocytes were exposed to an ischemia mimetic solution for 90 min and then reperfused with a normal solution for 30 min. 5-(N,N-dimethyl)-amiloride (DMA), a blocker of the Na(+)/H(+)exchanger, was administered during ischemia and the first 3 min of reperfusion or only during the first 3 min of reperfusion. Administration of DMA only upon reperfusion resulted in increased cell survival (81+/-1%, P<0.05) compared to using the drug during ischemia and reperfusion (63+/-3%) and in the absence of drug (60+/-1%). During ischemia, pH(i)was lower when DMA was present in the ischemic solution. The inhibition of the Na(+)/H(+)exchanger retarded the recovery of pH during reperfusion. The highest recovery of active cell shortening was observed when DMA was used at the beginning of reperfusion. The use of DMA also reduced the level of passive cell shortening during reperfusion, and when used at the beginning of reperfusion significantly increased the recovery of Ca(2+)transients. Our results demonstrate that the exchanger is primarily active during reperfusion and that inhibition of the exchanger solely at this time has a strong cardioprotective effect. PMID- 11013134 TI - Isolated 2-methylbutyrylglycinuria caused by short/branched-chain acyl-CoA dehydrogenase deficiency: identification of a new enzyme defect, resolution of its molecular basis, and evidence for distinct acyl-CoA dehydrogenases in isoleucine and valine metabolism. AB - Acyl-CoA dehydrogenase (ACAD) defects in isoleucine and valine catabolism have been proposed in clinically diverse patients with an abnormal pattern of metabolites in their urine, but they have not been proved enzymatically or genetically, and it is unknown whether one or two ACADs are involved. We investigated a patient with isolated 2-methylbutyrylglycinuria, suggestive of a defect in isoleucine catabolism. Enzyme assay of the patient's fibroblasts, using 2-methylbutyryl-CoA as substrate, confirmed the defect. Sequence analysis of candidate ACADs revealed heterozygosity for the common short-chain ACAD A625 variant allele and no mutations in ACAD-8 but a 100-bp deletion in short/branched chain ACAD (SBCAD) cDNA from the patient. Our identification of the SBCAD gene structure (11 exons; >20 kb) enabled analysis of genomic DNA. This showed that the deletion was caused by skipping of exon 10, because of homozygosity for a 1228G-->A mutation in the patient. This mutation was not present in 118 control chromosomes. In vitro transcription/translation experiments and overexpression in COS cells confirmed the disease-causing nature of the mutant SBCAD protein and showed that ACAD-8 is an isobutyryl-CoA dehydrogenase and that both wild-type proteins are imported into mitochondria and form tetramers. In conclusion, we report the first mutation in the SBCAD gene, show that it results in an isolated defect in isoleucine catabolism, and indicate that ACAD-8 is a mitochondrial enzyme that functions in valine catabolism. PMID- 11013135 TI - General equations for Pt, Ps, and the power of the TDT and the affected-sib-pair test. AB - Several equations are highlighted here, whose algebraic symmetries and generality make them very useful for understanding and comparing the properties of the transmission disequilibrium test (TDT) and affected sib-pair test. Methods using the equations are also presented that yield precise estimates of sample sizes needed for genome scans or for testing a single candidate gene, and these power methods are shown to compare favorably with alternative approaches recently described by Knapp (1999) and by Tu and Whittemore (1999). Simple relationships are also noted that summarize the relative sample sizes required for equivalent power to detect association by the TDT or case-control designs. As single nucleotide polymorphism (SNP) maps revolutionize the search for disease-causing genes, the equations should prove useful for planning and evaluating studies of linkage and association across a broad range of possible disease models and relationships between markers and linked disease loci. PMID- 11013136 TI - Mutations of the SCO1 gene in mitochondrial cytochrome c oxidase deficiency with neonatal-onset hepatic failure and encephalopathy. AB - Cytochrome c oxidase (COX) catalyzes both electron transfer from cytochrome c to molecular oxygen and the concomitant vectorial proton pumping across the inner mitochondrial membrane. Studying a large family with multiple cases of neonatal ketoacidotic comas and isolated COX deficiency, we have mapped the disease locus to chromosome 17p13.1, in a region encompassing two candidate genes involved in COX assembly-namely, SCO1 and COX10. Mutation screening revealed compound heterozygosity for SCO1 gene mutations in the patients. The mutated allele, inherited from the father, harbored a 2-bp frameshift deletion (DeltaGA; nt 363 364) resulting in both a premature stop codon and a highly unstable mRNA. The maternally inherited mutation (C520T) changed a highly conserved proline into a leucine in the protein (P174L). This proline, adjacent to the CxxxC copper binding domain of SCO1, is likely to play a crucial role in the tridimentional structure of the domain. Interestingly, the clinical presentation of SCO1 deficient patients markedly differs from that of patients harboring mutations in other COX assembly and/or maturation genes. PMID- 11013137 TI - Cloning of the gene encoding a novel integral membrane protein, mucolipidin-and identification of the two major founder mutations causing mucolipidosis type IV. AB - Mucolipidosis type IV (MLIV) is an autosomal recessive lysosomal storage disorder characterized by severe psychomotor retardation and ophthalmologic abnormalities, including corneal opacity, retinal degeneration, and strabismus. Unlike the situation in other lysosomal disorders, the accumulation of heterogeneous storage material observed in MLIV does not result from a block in the catabolic pathways but is due to an ill-defined transport defect in the late steps of endocytosis. With the aim of cloning the MLIV gene, we searched in the 19p13.2-13.3 region, where the locus previously had been assigned by linkage mapping. In this region, we have identified a novel gene that is mutated in all patients with MLIV who were enrolled in our study. One patient was homozygous for the splice-acceptor mutation, and another was homozygous for a deletion removing the first six exons of the gene. In addition, four compound heterozygotes for these two mutations were identified. Haplotype analysis indicates that we have identified the two major founder mutations, which account for >95% of MLIV chromosomes in Ashkenazi Jewish patients. The gene, ML4, encodes a protein named "mucolipidin, " which localizes on the plasma membrane and, in the carboxy-terminal region, shows homologies to polycystin-2, the product of the polycystic kidney disease 2 gene (PKD2) and to the family of transient receptor potential Ca(2+) channels. Mucolipidin is likely to play an important role in endocytosis. PMID- 11013145 TI - [Recent transmission of tuberculosis in Madrid (Spain): usefulness of molecular techniques]. AB - BACKGROUND: Recent studies suggest that many tuberculosis cases in urban areas result from recent transmission. The aim of this study was to determine patterns of tuberculosis transmission in Madrid. PATIENTS AND METHODS: A prospective population-based molecular epidemiological study of patients diagnosed of tuberculosis was conducted in three urban districts of Madrid (455.050 inhabitants) during 1997-1998. Clinical, demographic and epidemiological data were reviewed. Patients were included in clusters when their isolates contained: a) six or more IS6110 bands in an identical pattern, or b) five or fewer IS6110 bands that matched identically and had an identical spoligotyping pattern. RESULTS: Of 207 positive-culture patients, 148 (71,5%) were DNA fingerprinted. A total of 18 clusters which included 62 patients (41,9%) were identified. Clusters contained between 2 and 12 cases. Risk factors for clustering included: age < 35 years (OR = 4,1, 95% CI: 1,9-8,9), injection drug use (OR = 4,7, 95% CI: 1,6 14,8), HIV infection (OR = 2,7, 95% CI: 1,1-6,8), and a history of imprisonment (OR = 2,9, 95% CI: 1,2-7,2). The epidemiological investigation identified connections among 27% of clustered patients. CONCLUSIONS: A high proportion of cases of tuberculosis in urban Madrid result from recent transmission. Molecular epidemiology studies give valuable information for urban tuberculosis control. PMID- 11013146 TI - [Transmission of tuberculosis in the prisons of Madrid]. AB - BACKGROUND: Prisons are recognized as high-risk settings for tuberculosis transmission. The purpose of this study was to describe the characteristics of prisoners with tuberculosis and to determine the extent of transmission of tuberculosis in the prison population of Madrid. PATIENTS AND METHODS: Patients with positive culture for Mycobacterium tuberculosis between 1997 and 1998 from the prison system of Madrid were included. The medical records for these patients were reviewed, and they were also interviewed. Patients were included in clusters when their isolates contained 1) six or more IS6110 bands in an identical pattern or 2) five or fewer IS6110 bands that matched identically and had an identical spoligotyping pattern. RESULTS: Culture-proven tuberculosis was diagnosed in 97 prisoners (case rate: 693 per 100.000 per year). Isolates from 73 (75%) patients were available for fingerprinting. The mean age of these patients was 33 yr (SD: 6,9), 92% were male, and 14% were immigrants. Seventy-one percent (71%) of patients were HIV-positive, 32% were active intravenous drug users and 70% nonintravenous drug users in the last two years. Primary drug resistance to isoniazid was 1,8%. Forty-one percent of prisoners (41%) were grouped in 9 clusters. Epidemiological links were found in 37% of clustered patients. Risk factors for clustering were not detected. CONCLUSION: The results suggest recent transmission of tuberculosis in the prisons of Madrid. It is essential that correctional facilities comply fully with the recomendations for prevention and control of tuberculosis. PMID- 11013147 TI - [Injury rate and incidence of accidents with biological risk among infirmary students]. AB - BACKGROUND: A study of the incidence and characteristics of biological accidents among infirmary students during their practicals at the hospital. MATERIAL AND METHOD: A retrospective study carried out at five centres by means of two questionnaires, one on the duration of the training and the rate of accidents and the other on the characteristics, precautions and ports exposure behaviour. RESULTS: Out of 397 students, 70,5% had accidents at a rate of 64% (CI 95%, 59 68). Of these, 15% were accidents with biological risk, the majority being jabs (39%) and splashes (32,5%). It is worth note that 49,2% occurred while putting away the material and 58% in the absence of any individual protective measures. One out of 8 accidents implied a biological risk. CONCLUSIONS: A very high rate of accidents was observed with important deficiencies in security. PMID- 11013148 TI - [Prevalence and incidence of hepatitis A in patients with hepatitis B and C]. AB - BACKGROUND: Patients with chronic hepatitis are in risk to acquire a fulminant hepatitis associated with hepatitis A virus superinfection. PATIENTS AND METHODS: Antibodies against hepatitis A were study in serum from 353 patients with chronic hepatitis B or C. RESULTS: The prevalence of IgG-HAV antibodies was 81% in chronic hepatitis C patients, and 77% in chronic hepatitis B patients. The presence of anti-HAV antibodies was related to the patients' age. None case of acute hepatitis A in chronic hepatitis patients was detected. CONCLUSIONS: The prevalence of anti-HAV antibodies is high in patients with chronic viral hepatitis but the incidence of the disease is low. Hepatitis A vaccination should do with previously screening. PMID- 11013149 TI - [Tuberculosis, recent transmission and prisons]. PMID- 11013150 TI - [Tools for the practice of evidence based medicine (I). Update in evidence based information resources for clinical practice]. PMID- 11013151 TI - [On the revalidation for doctors in Spain]. PMID- 11013152 TI - [Diagnosis and treatment of myasthenia gravis]. PMID- 11013153 TI - [Abdominal pain and diarrhoea in a 33 year-old man]. PMID- 11013154 TI - [Consensus document on the prevention and control of tuberculosis in Spain]. PMID- 11013155 TI - [Methodological considerations about the prevalence of eating disorders]. PMID- 11013156 TI - [Reply] [In Process Citation] PMID- 11013157 TI - [Neuroleptic malignant syndrome and atypical antipsychotics]. PMID- 11013158 TI - [ [In Process Citation] PMID- 11013159 TI - Author's reply PMID- 11013160 TI - Low-cost high-volume ECCE with PC I0L PMID- 11013161 TI - Low-cost high-volume ECCE with PC I0L. AB - The following two letters address an article that appeared in the October 1999 issue of the Journal: Ruit S, Tabin GC, et al. Low-cost high-volume extracapsular cataract extraction with posterior chamber intraocular lens implantation in Nepal. (Ophthalmology 1999;106:1887-92) PMID- 11013163 TI - Author's reply PMID- 11013162 TI - Carotid artery compression of the optic nerve. PMID- 11013165 TI - Authors' reply PMID- 11013164 TI - Author's reply PMID- 11013166 TI - Long-term follow-up of retinal detachment from macular hole. PMID- 11013167 TI - LASIK after penetrating keratoplasty. PMID- 11013168 TI - Study of the AMO ARRAY silicone lens and a monofocal IOL. PMID- 11013170 TI - Author's reply PMID- 11013169 TI - Detection of choroidal calcium by CT scan. PMID- 11013171 TI - Corneal thickness and measured IOP. PMID- 11013172 TI - Author's reply PMID- 11013173 TI - Intracameral lidocaine in routine phacoemulsification. PMID- 11013174 TI - Authors' reply PMID- 11013175 TI - Author's reply PMID- 11013176 TI - Accurate intraocular pressure measurement-the myth of modern ophthalmology? PMID- 11013177 TI - If intraocular pressure measurement is only an estimate-then what? PMID- 11013178 TI - Mapping the visual field to the optic disc in normal tension glaucoma eyes. AB - PURPOSE: To establish the anatomical relationship between visual field test points in the Humphrey 24-2 test pattern and regions of the optic nerve head (ONH) DESIGN: Cross-sectional study. PARTICIPANTS: Glaucoma patients and suspects from the Normal Tension Glaucoma Clinic at Moorfields Eye Hospital. METHODS: Sixty-nine retinal nerve fiber layer (RNFL) photographs with well-defined RNFL defects and/or prominent bundles were digitized. An appropriately scaled Humphrey 24-2 visual field grid and an ONH reference circle, divided into 30 degrees sectors, were generated digitally. These were superimposed onto the RNFL images. The relationship of visual field test points to the circumference of the ONH was estimated by noting the proximity of test points to RNFL defects and/or prominent bundles. The position of the ONH in relation to the fovea was also noted. MAIN OUTCOME MEASURES: The sector at the ONH corresponding to each visual field test point, the position of the ONH in relation to the fovea, and the effect of the latter on the former. RESULTS: A median 22 (range, 4-58), of a possible 69, ONH positions were assigned to each visual field test point. The standard deviation of estimations was 7.2 degrees. The position of the ONH was 15.5 degrees (standard deviation 0.9 degrees ) nasal and 1.9 degrees (standard deviation 1.0 degrees ) above the fovea. The location of the ONH had a significant effect on the corresponding position at the ONH for 28 of 52 visual field test points. CONCLUSIONS: A clinically useful map that relates visual field test points to regions of the ONH has been produced. The map will aid clinical evaluation of glaucoma patients and suspects, as well as form the basis for investigations of the relationship between retinal light sensitivity and ONH structure. PMID- 11013179 TI - Accuracy of clinical estimates of intraocular pressure in Chinese eyes. AB - PURPOSE: To investigate the validity of clinical estimates of intraocular pressure (IOP) in Chinese people. DESIGN: Prospective, cross-sectional, hospital based in vivo study. PARTICIPANTS: Twenty-three ethnic Chinese adults (aged 35-82 years) undergoing routine phacoemulsification surgery were examined. TESTING: "True" IOP was measured with a solid-state hemodynamic monitor through a cannula in the anterior chamber. IOP was set successively to 10, 20, and 30 mmHg in each subject, using a reservoir of balanced salt solution. Intraocular pressure was simultaneously estimated by use of a hand-held applanation tonometer (Perkin's) and a Tono-Pen. The association between ocular biometric variables and measurement error was examined. MAIN OUTCOME MEASURES: The median of three readings at each IOP level was taken as the IOP estimate of each instrument. Measurement error was calculated as the mean difference (tonometer minus direct measurement). RESULTS: The error for the hand-held applanation tonometer was 1.6, -4.3 and -5.7 at 10, 20, and 30 mmHg, respectively. For the Tono-Pen the measurement error was +0.4, -2.0, and -4.1 at 10, 20, and 30 mmHg, respectively. We could identify no association between measurement error and corneal thickness or curvature, anterior chamber depth, or axial length. CONCLUSIONS: The applanation tonometer and Tono-Pen underestimate the true IOP in Chinese eyes. Error increases as true IOP increases. These tonometers do not give an accurate estimate of IOP in East Asians. PMID- 11013180 TI - Long-term follow-up of trabeculectomy with intraoperative 5-fluorouracil for uveitis-related glaucoma. AB - PURPOSE: To determine the effect of intraoperative application of 5-fluorouracil (5-FU) on the long-term outcome of trabeculectomy in uveitis-related glaucoma. DESIGN: An open, prospective, noncomparative case study to evaluate the use of intraoperative application for 5 minutes of 25 mg/ml 5-FU in primary trabeculectomy for uveitis-related glaucoma. PARTICIPANTS: Fifty eyes of 43 consecutive patients were enrolled in the study. METHODS: Data were recorded prospectively on specifically designed forms at visits every 3 months after surgery. MAIN OUTCOME MEASURES: The intraocular pressure, need for additional medication, and the need for repeat surgery were the criteria that determined the outcome of surgery. RESULTS: No significant intraoperative, postoperative, or late complications of 5-FU administration were noted. Successful filtration was achieved in 82% of eyes at 1 and 2 years after surgery. The success rate was 67% at 5 years. Failure was significantly more common and occurred earlier in patients of black ethnic origin. No deleterious effect on control of uveitis in relation to surgical intervention was observed. CONCLUSIONS: Intraoperative application of 5-FU appears to be a long-term, safe, and effective adjunct to trabeculectomy in uveitis-related glaucoma. PMID- 11013181 TI - Management of alkali burns : an 11-year retrospective review. AB - OBJECTIVE: To review the spectrum of patients with alkali burns admitted over an 11-year period and to assess the clinical outcomes after the introduction of a standard alkali burn treatment protocol. DESIGN: Retrospective nonrandomized comparative study. PATIENTS AND INTERVENTIONS: A total of 121 patient records with alkali burns (n = 177 eyes) admitted to a tertiary hospital between 1987 and 1998 were reviewed. Eyes treated with a standard alkali burn treatment protocol, which included intensive topical steroids, ascorbate, citrate, and antibiotics, were compared with eyes treated by conservative management with antibiotics, and a short course of steroids. MAIN OUTCOME MEASURES: Time to corneal reepithelialization, final best-corrected visual acuity, and time to visual recovery, length of hospital stay, and complications were analyzed. RESULTS: The standard protocol tended to delay corneal reepithelialization by one day (P: = not significant) in eyes with grade 1 burns (n = 76) and by 2 days (P: = 0.04) in grade 2 burns (n = 52), with no difference in final visual outcome. There were 37 eyes with grade 3 burns. Those treated with the standard protocol showed a trend toward more rapid corneal reepithelialization. Twenty-seven of 29 (93%) eyes with grade 3 injuries achieved a final best-corrected visual acuity of 20/40 or better compared with 3 of 6 (50%) eyes not treated according to the standard protocol (P: = 0.02). Eyes with grade 4 burns (n = 12), whether treated with the standard protocol or not, required 10 to 12 weeks for corneal reepithelialization. There was no statistically significant difference in final visual acuity. CONCLUSIONS: On the basis of our findings, a number of recommendations can be made for the management of alkali injuries. Patients with a grade 1 or 2 injury do not require routine admission and do not benefit from the use of intensive treatment with ascorbate and citrate. A trend toward more rapid healing and a better final visual outcome were apparent in grade 3 burns, but our standard protocol made no difference in grade 4 burns. PMID- 11013182 TI - Surgical treatment of advanced pellucid marginal degeneration. AB - PURPOSE: To determine the efficacy of simultaneous peripheral crescentic lamellar keratoplasty (LK) and central penetrating keratoplasty (PK) for advanced pellucid marginal degeneration (PMD). DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Five patients with advanced PMD. METHOD: Simultaneous peripheral crescentic LK and central PK followed by selective suture removal and astigmatic keratotomy in the postoperative period. MAIN OUTCOME MEASURES: These included interval of time required for visual rehabilitation, best spectacle-corrected visual acuity achieved, and amount of corneal astigmatism, as measured by videokeratography. Measuring the change in corneal astigmatism by videokeratography over the subsequent follow-up period after spectacle correction had been prescribed assessed stability of the achieved refraction. RESULTS: Visual acuity results were 20/40 in three eyes, 20/80 in one eye, and 20/400 in one eye. The latter two had decreased acuity from posterior subcapsular cataract formation, which may have been caused by topical steroid use. The time required for visual rehabilitation ranged from 5.13 to 10.93 (mean, 9.92) months, and the amount of corneal astigmatism at the end of this period ranged from 0.3 diopters (D) to 5.3 D. A tendency for an increase in "with the rule astigmatism" after the rehabilitation period was noted. Two patients had elevations of intraocular pressure that responded to reduction in topical steroid dose. CONCLUSIONS: The short-term results with this technique are excellent in that it provides early and stable visual rehabilitation in patients with advanced PMD. Low to moderate levels of postkeratoplasty astigmatism were achieved in all the eyes treated. The usual tendency of an increase in "against the rule astigmatism" that occurs when PK alone is done for PMD was eliminated. PMID- 11013183 TI - Flow cytometric analysis of conjunctival epithelium in ocular rosacea and keratoconjunctivitis sicca. AB - PURPOSE: To investigate by flow cytometry and impression cytology (IC) specimens the inflammatory status of the conjunctival epithelium and goblet cell density in two series of patients with rosacea and dry eye syndrome compared with a population of healthy subjects. DESIGN: Nonrandomized, prospective, comparative case series. PARTICIPANTS: Twenty-six eyes of 13 patients with rosacea, 26 eyes of 13 patients with dry eye syndrome, and 24 eyes of 12 control subjects were included in this study. METHODS: IC specimens were collected after clinical examination of the ocular surface and analyzed by flow cytometry, using antibodies directed to human lymphocyte antigen-DR (HLA-DR), intercellular adhesion molecule-1 (ICAM-1) (CD 54), and the peptidic core of the conjunctival mucin (M1/MUC5AC). The percentage of positive cells was calculated and levels of fluorescence expression quantified and compared with those obtained in a series of 12 healthy subjects. MAIN OUTCOME MEASURES: Tear break-up time (TBUT), Schirmer test, fluorescein and lissamin green stainings, and IC were realized in this study. RESULTS: A significant increase of HLA-DR and ICAM-1 expressions by epithelial cells was consistently found in the two pathologic groups compared with levels calculated in normal eyes. The two markers were well correlated with each other and inversely with TBUT and Schirmer test. The percentage of goblet cells was significantly decreased in rosacea patients and in dry eye patients compared with the normal group with a significant negative correlation with both HLA DR and ICAM-1 markers. CONCLUSIONS: Ocular rosacea and keratoconjunctivitis sicca were associated with severe ocular surface changes, such as an overexpression of inflammatory markers and a significant decrease in the number of goblet cells. PMID- 11013185 TI - Discussion by herbert E. Kaufman, MD PMID- 11013184 TI - Preliminary clinical results of posterior lamellar keratoplasty through a sclerocorneal pocket incision. AB - PURPOSE: To report the preliminary results of a surgical technique for transplantation of posterior corneal tissue through a sclerocorneal pocket incision for corneal endothelial disorders. DESIGN: Retrospective, noncomparative, interventional cases series. PARTICIPANTS AND INTERVENTION: In seven sighted human eyes, a deep stromal pocket was created across the cornea through a 9.0-mm superior scleral incision. A 7.0- or 7.5-mm diameter, posterior lamellar disc was excised and replaced by a 'same size' donor posterior disc, without suture fixation. The scleral incision was sutured. MAIN OUTCOME MEASURES: Intra- and postoperative complications, best spectacle-corrected visual acuity, keratometry, topography, biomicroscopy, pachymetry, and endothelial cell density were evaluated. RESULTS: Six to 12 months after surgery, all transplants were clear and in position. Best spectacle-corrected visual acuity was limited by preexisting maculopathies in two eyes and varied from 20/80 to 20/20. Postoperative astigmatism averaged 1. 54 diopters (D; standard deviation [SD] +/- 0.81 D), pachymetry averaged 0.49 mm (SD +/- 0.09 mm), and postoperative endothelial cell density averaged 2520 cells/mm(2) (SD +/- 340 cells/mm(2)). In one eye, a microperforation occurred during stromal pocket dissection so that the procedure was converted into a penetrating keratoplasty. CONCLUSIONS: Posterior lamellar keratoplasty through a sclerocorneal pocket incision is a feasible surgical approach to manage corneal endothelial disorders. PMID- 11013187 TI - Discussion by stephen trokel, MD PMID- 11013186 TI - Early results of hyperopic and astigmatic laser in situ keratomileusis in eyes with secondary hyperopia. AB - PURPOSE: To assess the safety and efficacy of laser in situ keratomileusis (LASIK) for secondary hyperopia and hyperopic astigmatism and to develop a VISX STAR S2 LASIK nomogram (VISX Inc., Santa Clara, CA) for consecutive hyperopia after prior myopic refractive surgery. DESIGN: Prospective, nonrandomized, self controlled interventional study. PARTICIPANTS: Thirty patients with consecutive hyperopia or hyperopia and astigmatism after LASIK, photorefractive keratectomy, automated lamellar keratoplasty, or radial keratotomy. INTERVENTION/METHODS: Prospective evaluation of LASIK in 30 secondary eyes with fogged manifest sphere from +0.5 to +6.0 diopters (D) and cylinder from 0 to +5.0 D. MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), and spherical equivalent (SE). RESULTS: Mean manifest SE was +1.73 +/- 0.79 D before surgery, -0.13 +/- 1.00 D at 6 months after surgery, and -0.18 +/- 1.08 D at 1 year after surgery. At 6 months, 84% of patients with secondary hyperopia had UCVA of 20/40 or better; 76% were within +/-1 D of emmetropia. At 1 year, 85% had UCVA of 20/40 or better and 85% were within +/-1 D of emmetropia. No patients with secondary hyperopia lost 2 or more lines of BCVA at 1 year. Complications included intraoperative bleeding (3.3%), intraoperative epithelial defect (3.3%), transient interface debris (3.3%), significant dry eye (3.3%), blood in interface (3.3%), irregular astigmatism (6.7%), slight decentration (6.7%), trace haze (6.7%), or mild epithelial ingrowth not requiring removal (3.3%). CONCLUSIONS: These early data suggest that LASIK for consecutive hyperopia from +0.5 to +5.50 D and astigmatism from 0 to +2.75 D using the VISX STAR S2 benefits from a nomogram adjusted for preoperative refraction, age, and prior refractive surgery, and is safe and effective. PMID- 11013188 TI - Contrast sensitivity evaluation after laser in situ keratomileusis. AB - OBJECTIVE: To determine the effects of laser in situ keratomileusis (LASIK) on best-corrected visual acuity (BCVA) and contrast sensitivity. DESIGN: Prospective, observational case series. PARTICIPANTS: One hundred twelve eyes, in 65 patients with myopia and myopia with astigmatism, who underwent LASIK. TESTING AND MAIN OUTCOME MEASURES: Best-corrected visual acuity using the Snellen visual acuity chart and contrast sensitivity using the CSV 1000 (Vector Vision, Dayton OH) was tested before surgery and 1 week, 1 month, and 3 months after surgery in patients who underwent LASIK. RESULTS: Contrast sensitivity was depressed for patient eyes with spherical equivalence (SE) between -1.25 diopters (D) and 13.75 D, at 12 cycles/degree for at least 3 months and at 18 cycles/degree for 1 week after LASIK. For patient eyes with SE between -1.25 D and -6.00 D, contrast sensitivity was depressed only at 12 cycles/degree for at least 3 months after LASIK. For patient eyes with SE between -6.00 D and -13.75 D, contrast sensitivity was depressed at 6, 12, and 18 cycles/degree 1 week after LASIK but returned toward preoperative levels by 1 month after surgery. Despite the slight decreases in contrast sensitivity, all scores were still within the range of normal values except for 12 cycles/degree for 3 months and 18 cycles/degree at 1 week after surgery in the high myopia group. Although highly myopic patients, compared with patients with low myopia, had slightly less BCVA before surgery, both groups maintained their preoperative BCVA at all postoperative visits. CONCLUSIONS: Based on this study, we conclude that LASIK has little effect on BCVA and contrast sensitivity for up to 3 months after surgery. PMID- 11013190 TI - Discussion by allan E. Kolker, MD PMID- 11013189 TI - Posterior capsular opacification in phacotrabeculectomy : a long-term comparative study of silicone versus acrylic intraocular lens. AB - PURPOSE: To investigate the long-term outcomes of silicone versus acrylic intraocular lens (IOL) implantation in phacotrabeculectomy (PT) with special emphasis on posterior capsular opacification. DESIGN: Long-term follow-up on prior 1-year prospective, randomized study. PARTICIPANTS: A total of 200 eyes of 200 consecutive primary open-angle glaucoma patients who had undergone primary PT with capsular bag implantation of either a silicone IOL (102 eyes) or an acrylic IOL (98 eyes) according to the initial short-term prospective, randomized study protocol. INTERVENTION: The study eyes underwent primary trabeculectomy, phacoemulsification, and posterior chamber IOL implantation. Adjunctive mitomycin C was used selectively, primarily in patients with one or more risk factors for filtration failure. MAIN OUTCOME MEASURES: Incidence of posterior capsular opacification (PCO), best-corrected visual acuity (BCVA), intraocular pressure (IOP), number of pressure-lowering medications, and filtration success rates, defined as maintenance of target IOP while on one (criteria 1) or zero (criteria 2) pressure-lowering medications without further surgical intervention. RESULTS: At 3-year follow-up, the PCO rate and BCVA did not differ significantly between the two groups (P: > 0.05 for both). In addition, there were no significant differences in IOP, number of medications, and filtration success rate between the two groups (P: > 0.05 for each). CONCLUSIONS: There were no significant long term differences between the silicone and acrylic IOL groups in PCO, BCVA, IOP, number of medications, and success of filtration surgery after PT. Both groups attained significant improvement in BCVA and IOP control after surgery. PMID- 11013191 TI - Pediatric orbital floor fracture : direct extraocular muscle involvement. AB - OBJECTIVE: To study the clinical presentation, operative findings, and postoperative results of a surgical series of isolated orbital floor fractures in children. DESIGN: Noncomparative, retrospective, consecutive case series. PARTICIPANTS: Thirty-four patients (34 orbits) less than 18 years of age with isolated orbital floor fractures. Indications for surgery were severe limitation of extraocular ductions, 22 of 34; enophthalmos, 8 of 34: or both, 4 of 34. INTERVENTION: Surgical repair. MAIN OUTCOME MEASURES: Cause of fracture, symptoms, clinical signs, radiographic data, operative findings, postoperative results, and complications. RESULTS: Children older than 12 years of age were more likely to sustain an orbital floor fracture as a result of interpersonal violence than were children less than 12 years of age (P: = 0.020). Sixty-two percent of patients (21 of 34) exhibited pain with eye movements and/or nausea and vomiting. Most had a trapdoor type fracture (21 of 34). The inferior rectus muscle was entrapped in the orbital floor fracture in 69% (18 of 26) of patients with a severe limitation of ocular ductions. Preoperative nausea and vomiting were immediately relieved after surgery. The median time for improvement of preoperative duction deficits and diplopia was 4 days for patients receiving surgery within 7 days and 10.5 days for those undergoing surgery after 14 days (P: = 0.030). Resolution of duction deficits or diplopia was not dependent on time of surgery if performed within 1 month of injury. Loss of vision, worsening of motility, or implant complications did not occur. CONCLUSIONS: Pediatric patients with isolated orbital floor fractures who had pain, nausea, vomiting, and severe limitation of extraocular motility often have direct entrapment of the inferior rectus muscle into the fracture site. Surgical repair rapidly relieved preoperative pain, nausea, and vomiting. For patients with severe limitation of ductions, early surgical repair within 7 days of injury resulted in more rapid improvement of ductions and diplopia than surgery performed later. PMID- 11013192 TI - Restoration of fusion in children with intracranial tumors and incomitant strabismus. AB - PURPOSE: Intracranial tumors may cause eye misalignment and interruption of sensory fusion. The ocular misalignment may be permanent or may be corrected after tumor treatment with or without specific strabismus treatment. This report analyzes the binocular vision outcome of children with misaligned eyes from brain tumors who regain orthotropia. DESIGN: A retrospective noncomparative case series. PARTICIPANTS: Twenty-three surviving children less than 18 years of age with a new heterotropia and absence of fusion associated with the development of a brain tumor. INTERVENTIONS: Tumor resection/radiation/chemotherapy and necessary strabismus management. MAIN OUTCOME MEASURE: Stereopsis (>/= 4 of 10 circles) measured with the Randot II stereo test at near fixation. RESULTS: Fourteen children regained orthotropia either after tumor therapy, strabismus treatment, or both. Ten of these 14 children with realigned vision regained high grade stereovision. Nine patients did not regain orthotropia and were excluded. The mean age at tumor diagnosis of the fusing group was 9.9 years (range, 3-17 years) compared with 8.5 years (range, 6-12 years) in the four realigned nonfusing patients. The mean duration of misalignment was 12 months (range, 2-51 months) for the fusing group, and 45 months (range, 14-120 months) for the nonfusing group. CONCLUSIONS: Children and adolescents with brain tumor associated eye misalignment may regain the ability to fuse if their misalignment can be corrected. An improved prognosis was noted for those patients when the misalignment had been present for a shorter duration. These data suggest that the outcome may be better for incomitant strabismus than that reported for acute comitant esotropia. PMID- 11013193 TI - Efficacy of diclofenac versus dexamethasone for treatment after strabismus surgery. AB - OBJECTIVE: To compare the efficacy of topical diclofenac sodium 0.1% versus dexamethasone 0.1% on the conjunctival healing process and on intraocular pressure (IOP) after strabismus surgery. DESIGN: A randomized clinical trial. PARTICIPANTS: Forty consecutive pediatric patients who underwent strabismus surgery. INTERVENTION: The patients were assigned before surgery to receive topical diclofenac 0.1% (study group, 20 patients) or dexamethasone 0.1% (control group, 20 patients) from immediately after surgery to up to 4 weeks after surgery (both combined with chloramphenicol 0.2%, polymyxin B sulfate 2500 U). MAIN OUTCOME MEASURES: Between-group comparison of five parameters: patient discomfort, conjunctival chemosis, inflammation, gap, and intraocular pressure (IOP) at 1, 2, and 4 weeks after surgery. RESULTS: At postoperative week 2, the diclofenac-treated group showed significantly less patient discomfort and less conjunctival inflammation, edema, and gap than the dexamethasone group (P: = 0.003, P: = 0.04, P: = 0.02, P: = 0. 001, respectively). At week 4, the study patients continued to show less discomfort and conjunctival gap (P: = 0.02). The dexamethasone group showed a significant change in IOP between the preoperative and the fourth postoperative week (P: = 0.001 in the right eye, P: = 0.0005 in the left eye) and an increased prevalence of higher IOP during the fourth postoperative week (P: = 0.01 in the right eye, P: = 0.02 in the left eye). Thirty-eight percent of the dexamethasone group showed an increase in IOP to more than 21 mmHg during the four postoperative weeks. No increase in IOP was noted in the diclofenac group. CONCLUSIONS: Topical diclofenac is superior to dexamethasone for each of the five postoperative parameters examined. Its maximal effect occurred at 2 weeks after surgery, without an increase in IOP or in local subconjunctival hemorrhage. PMID- 11013194 TI - Comparison of artificial eye amplitudes with acrylic and hydroxyapatite spherical enucleation implants. AB - PURPOSE: To compare artificial eye amplitudes in patients who randomly received either a hydroxyapatite or an acrylic, scleral-covered spherical implant after enucleation. DESIGN: Randomized, controlled trial. PARTICIPANTS: Thirty-four consecutive patients who underwent enucleation because of an intraocular melanoma and 21 healthy control participants from the hospital staff. METHODS: Eligible patients randomly received a hydroxyapatite or an acrylic, scleral-covered spherical orbital implant. Fourteen patients were fitted with a hydroxyapatite implant, and 16 were fitted with an acrylic implant. We measured horizontal and vertical saccadic amplitudes of both the artificial eye and the healthy eye. Measurements were performed with the magnetic search coils technique. Saccadic amplitudes of the artificial eye were compared with the healthy eye of the patient. The amplitudes of the healthy eyes were compared with saccadic amplitudes of control participants. The interval from surgery to measurements was at least 3 months in all patients. Saccadic gain (artificial eye and eye amplitude divided by target amplitude) and saccadic symmetry (artificial eye amplitude divided by healthy eye amplitude) were calculated. MAIN OUTCOME MEASURES: Saccadic gain and saccadic symmetry. RESULTS: The gain in the healthy eyes of the patients was comparable with the gain of the control eyes. Saccadic symmetry was 1.0 in control participants. In patients, it was 0.334 in horizontal saccades and 0.577 in vertical saccades. However, saccadic symmetry did not differ significantly between the acrylic group and the hydroxyapatite group (P: > 0.1 for any saccadic direction). Equivalence was detectable with a power more than 90% for horizontal saccades and more than 80% for vertical saccades. Curvilinearity was rejected for both patient groups and for all saccadic directions (P: > 0.5). CONCLUSIONS: When no motility peg is placed, acrylic and hydroxyapatite spherical implants yield comparable saccadic amplitudes of the artificial eye. Artificial eye amplitudes were markedly more restricted horizontally than vertically. In all saccadic directions, the relation between target amplitude and artificial eye amplitude was linear. PMID- 11013195 TI - Eyelid abnormalities in lamellar ichthyoses. AB - PURPOSE: To describe eyelid abnormalities in lamellar ichthyoses (LI). DESIGN: Retrospective observational case series. PARTICIPANTS: Eight patients with classic LI and two patients with congenital ichthyosiform erythroderma. METHODS: Results of eyelid and corneal examinations of 10 patients with LI were reviewed and analyzed with emphasis on the relationship between eyelid ectropion and corneal damage. RESULTS: All patients presented with cicatricial lagophthalmos. Of the eight patients with classic LI, five had ectropion of the four eyelids, one had only lower ectropion, and two had no degree of ectropion. Two patients with congenital ichthyosiform erythroderma had distinct eyelid abnormalities, including madarosis and eyelash retraction. Loss of vision caused by corneal damage was found in three patients with classic LI. Of these three patients, two did not have upper eyelid ectropion. CONCLUSIONS: Severe corneal damage can occur in LI even if there is no upper or lower eyelid ectropion. PMID- 11013196 TI - Computed tomography in the diagnosis and prognosis of open-globe injuries. AB - OBJECTIVE: Determine sensitivity, specificity, and prognostic signs of orbital and ocular computed tomography (CT) in diagnosing patients with open globe injury. DESIGN: Randomized masked review of computed tomograms and retrospective clinical correlation of patients with ocular trauma. PARTICIPANTS: Two hundred patients who underwent CT evaluation for ocular trauma between 1989 and 1993. METHODS: CTs were read by three masked observers; findings were tabulated and compared for variability among observers; sensitivity and specificity were calculated and CT findings were grouped according to visual outcome retrieved from record review. RESULTS: In the absence of clinical information, sensitivity and specificity were 75% and 93%, respectively. The positive predictive value ranged from 88% to 97%, with a calculated overall positive predictive value of 95%. Patients who had a poor visual outcome (visual acuity <2/200) or who underwent enucleation had significantly more CT findings than patients with a good visual outcome. Vitreous hemorrhage, absence of lens, and severe distortion of vitreous space are among the most common CT findings associated with poor visual outcome. CONCLUSIONS: CT is not sensitive enough to be solely relied upon for diagnosis of all open globe injuries. CT findings only complement clinical findings, increasing the clinician's overall ability to make an accurate diagnosis of open globe injury, and may provide useful prognostic information regarding visual outcome. PMID- 11013197 TI - Pseudotumor cerebri and optic nerve sheath decompression. AB - OBJECTIVE: To determine the efficacy and safety of optic nerve sheath decompression in a large population of patients with pseudotumor cerebri with visual loss despite medical treatment and to suggest a treatment algorithm on the basis of these data. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: One hundred fifty-eight eyes in 86 patients with pseudotumor cerebri. INTERVENTION: Optic nerve sheath decompression. MAIN OUTCOME MEASURES: Visual acuity, visual fields, and surgical complications. RESULTS: After optic nerve sheath decompression for pseudotumor cerebri, visual acuity stabilized or improved in 148 of 158 (94%) eyes, and visual fields stabilized or improved in 71 of 81 (88%) eyes. Surgical complications, most of which were transient and benign, were seen in 39 of 86 patients. Only one eye in one patient had permanent severe visual loss secondary to an operative complication. CONCLUSIONS: In patients with pseudotumor cerebri with progressive visual loss despite maximum medical therapy, optic nerve sheath decompression is a safe and effective means of stabilizing visual acuity and the visual fields of those tested. PMID- 11013199 TI - Discussion by H. Stanley thompson, MD PMID- 11013198 TI - Portable pupillography of the swinging flashlight test to detect afferent pupillary defects. AB - OBJECTIVE: To investigate the ability of a portable, personal computer-driven, pupillometer to record the pupillary response curve during the swinging flashlight test. Also, to determine whether these response curves can be used to identify and quantify relative asymmetry in the pupillary light reflex between eyes in healthy volunteers with simulated afferent pupil defects (APDs) and patients with optic neuropathies. DESIGN: Comparative, observational case series and instrument validation. PARTICIPANTS: Healthy volunteers with no known ocular disease and patients (n = 20) with various optic neuropathies noted to have relative APDs on examination. METHODS: Pupillary response curves of the right eye were recorded with a portable, electronic, infrared pupillometer from healthy volunteers (with and without simulated APDs) and patients with APDs while the light stimulus alternated between eyes, simulating the swinging flashlight test. Simulated APDs in healthy volunteers were created with increasingly dense neutral density filters in front of the left eye. MAIN OUTCOME MEASURES: Differences in constriction amplitude, latency, and constriction velocity of the pupillary response with right eye stimulation versus left eye stimulation in both groups of subjects. RESULTS: A significant correlation between neutral density filter strength and intereye differences was seen for all measurement parameters in volunteers with simulated APDs. Depending on the measurement parameter and stimulus intensity, simulated APDs of 0.6 log units or more could be distinguished from normal responses. Clinically graded true APDs had intereye differences similar to simulated APDs of the same density. Those with real and simulated APDs of 0.9 log units or more could be distinguished from healthy volunteers with 80% sensitivity and 92% specificity. Responses from those with real and simulated small APDs of 0.3 to 0.6 log units could not be distinguished reliably. CONCLUSIONS: Portable, personal-computer driven, electronic, infrared pupillography can record the swinging flashlight test accurately and identify large afferent pupillary defects. An affordable, portable, reliable device for identifying relative APDs would be useful in the identification and follow-up of patients with neurogenic vision loss. PMID- 11013200 TI - Primary scleral buckling in open-globe injury involving the posterior segment. AB - PURPOSE: To determine whether scleral buckle placement at the time of primary repair of open-globe injury of the posterior segment is beneficial. DESIGN: Retrospective, comparative, nonrandomized interventional study. PARTICIPANTS: One hundred twenty-five open-globe injuries treated at the Duke University Medical Center from June 1980 to May 1997. METHODS: Open-globe injuries were classified with the Open-globe Injury Classification. Eyes that had zone 2 and 3 injuries that had a primary buckle placed were compared with those that did not. MAIN OUTCOME MEASURES: Subsequent retinal detachment, visual outcome, and need for subsequent scleral buckling. RESULTS: The rate of retinal detachment and the visual outcome were similar in the two groups. More than half of those who did not have a primary buckle placed had subsequent scleral buckling surgery. CONCLUSIONS: Many open-globe injuries of the posterior segment require eventual scleral buckle. There may be a role for placement of a scleral buckle at the time of primary repair. PMID- 11013201 TI - Radioactive plaque therapy for metastatic choroidal carcinoma. AB - PURPOSE: To describe the outcome of radioactive episcleral plaque therapy for treatment of metastatic carcinoma to the choroid. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Five patients (six eyes) with carcinoma metastatic to the choroid. METHODS: Retrospective review of the clinical records of five patients (six eyes) who underwent radioactive episcleral plaque therapy for choroidal metastases. MAIN OUTCOME MEASURES: Tumor height, visual acuity, radiation optic neuropathy, and radiation retinopathy. RESULTS: Radioactive episcleral plaque therapy resulted in shrinkage of the treated tumors and resolution of subretinal fluid in all eyes. After plaque treatment, best corrected visual acuity was maintained within two lines of initial visual acuity for two eyes, decreased more than two lines for one eye, and improved more than two lines in three eyes. The treatment was well tolerated and there was no acute toxicity. Late complications included optic nerve atrophy (at 2 years) with proliferative radiation retinopathy (at 3 years) in one eye and optic atrophy (at 6 months) in another eye that had received prior external beam therapy. CONCLUSIONS: In carefully selected cases, radioactive episcleral plaque therapy appears to be an effective and reasonable treatment for carcinoma metastatic to the choroid. PMID- 11013202 TI - Dexamethasone concentration in the subretinal fluid after a subconjunctival injection, a peribulbar injection, or an oral dose. AB - PURPOSE: To determine dexamethasone concentrations in the subretinal fluid of patients after a peribulbar injection, a subconjunctival injection, or an oral dose of dexamethasone and to compare the results with those of previous similar studies of dexamethasone concentrations in the vitreous. DESIGN: Prospective, nonrandomized, comparative trial. PARTICIPANTS: One hundred forty-eight patients with a rhegmatogenous retinal detachment. METHODS: Fifty patients received a peribulbar injection of 5 mg dexamethasone disodium phosphate, 49 received a subconjunctival injection of 2.5 mg dexamethasone disodium phosphate, and 49 received an oral dose of 7. 5 mg dexamethasone at various time intervals before surgery. At the time of surgery, a subretinal fluid sample was taken from each patient. MAIN OUTCOME MEASURES: The dexamethasone concentration in the subretinal fluid measured by radioimmunoassay. RESULTS: The estimated maximum dexamethasone concentrations in the subretinal fluid after the peribulbar injection, the subconjunctival injection, and the oral dose were, respectively, 82.2 ng/ml (standard error, 17. 6), 359 ng/ml (standard error, 80.2), and 12.3 ng/ml (standard error, 1.61). Corrected for dose, the maximum dexamethasone concentrations after subconjunctival injection and peribulbar injection were, respectively, 120 (95% confidence interval, 54/180) and 13 (95% confidence interval, 6.8/20) times greater than after oral administration. CONCLUSIONS: A subconjunctival injection of dexamethasone disodium phosphate is more effective in delivering dexamethasone into the subretinal fluid of patients with a rhegmatogenous retinal detachment compared with peribulbar injection or oral administration. The subretinal dexamethasone concentrations were higher than concentrations measured in the vitreous in previous studies with a similar setup after all three delivery methods. PMID- 11013204 TI - Discussion by raymond R. Margherio, MD PMID- 11013203 TI - Macular hole surgery with and without internal limiting membrane peeling. AB - OBJECTIVE: To compare results of surgery for idiopathic macular hole with and without internal limiting membrane (ILM) peeling in a series of consecutive patients over a 5-year period. DESIGN: A retrospective, nonrandomized, comparative trial with concurrent control group. PARTICIPANTS: Forty-four eyes with macular holes of less than or equal to 6 months duration without ILM peeling were compared to 116 eyes with ILM peeling and the same hole duration. A third group of 65 eyes with ILM peeling and duration greater than 6 months was also evaluated. INTERVENTION: All eyes underwent pars plana vitrectomy with or without ILM peeling, intravitreous gas, and positioning face down. No adjunctive therapies were used in any group. MAIN OUTCOME MEASURES: Comparing the closure and/or reopening rate, prognosis, visual acuity, and complications for macular holes with and without ILM peeling. RESULTS: All patients had postsurgical follow up of 18 months or greater. Primary closure was significantly improved with ILM peeling with 116 of 116 eyes (100%) showing no reopenings versus 36 of 44 holes (82%) primarily closed, 9 of which (25%) reopened without ILM peeling (P: < 0.00001) in holes less than or equal to 6 months. The 27 eyes without ILM peeling that had successful surgery displayed a mean postoperative vision of 20/40, which is the same as the successful eyes with ILM peeling (P: = 0.6). The 52 stage II eyes with ILM peeling had a mean postoperative vision of 20/30, and 48 of the 52 eyes (92%) were 20/40 or better. Stage III eyes (greater than 400-microm holes) without ILM peeling had a poor prognosis, with 6 of the 25 eyes (24%) having initial surgery fail and an additional 4 of 25 eyes (16%) reopening. Without ILM peeling, holes less than 300 microm had only one reopen, whereas holes greater than or equal to 300 microm had 16 of the 17 (94%) primary failures and/or reopenings (P: < 0.001). All 12 holes that reopened and/or primarily failed were repaired with ILM peeling with excellent visual recovery. Macular holes with a duration greater than 6 months were treated with ILM peeling, and 63 of 65 holes (97%) were closed primarily and 65% had an increase in vision by two or more Snellen lines. CONCLUSIONS: ILM peeling significantly improves visual and anatomic success in all stages of recent and chronic macular holes and reopened and failed holes, while eliminating reopening for holes greater than 300 microm. PMID- 11013205 TI - Effect of stimulus size on static visual fields in patients with retinitis pigmentosa. AB - PURPOSE: To determine the effect of stimulus size on sensitivity of patients with retinitis pigmentosa (RP) as measured by automated static perimetry. DESIGN: Comparative case series. PARTICIPANTS: Thirty-nine patients with RP and a control group of 10 healthy volunteers. METHODS: Automated static perimetry (full threshold programs 24-2 or 30-2) was performed twice on one eye of each participant using stimulus sizes III (0.43 degrees diameter) and V (1.72 degrees diameter). Data from the same 50 test locations were used from each field. MAIN OUTCOME MEASURES: At each location, for each participant, the size effect was computed as the difference (in decibels) in sensitivities for sizes V and III, and the average sensitivity was computed as the mean of sensitivities for the two sizes. RESULTS: For both patient and control groups, the size effect was negatively correlated with average sensitivity (r(2) > 0.124; P: < 0.001). The mean size effect was significantly greater for the patient group than for the control group: 8.6 (+/- 3.6) dB versus 5. 4 (+/- 2.2) dB (t = 18.0; P: < 0.001). The percentage of abnormal locations (more than 8 dB below mean normal) tended to be lower for size V than for size III, with a mean of 67% for size V versus 95% for size III. The percentage of absolute defects was also lower for size V than for size III, with a mean of 35% for size V versus 54% for size III. CONCLUSIONS: In damaged regions of the visual fields of patients with RP, increase in stimulus size from III to V can produce abnormally large increases in perimetric sensitivity. Size III may be more useful than size V for detection of field abnormality, whereas size V may be more useful than size III for observing progression of advanced RP. PMID- 11013206 TI - Ocular-central nervous system lymphoma mimicking posterior scleritis with exudative retinal detachment. AB - OBJECTIVE: We describe an unusual ocular presentation of ocular-central nervous system lymphoma in a young patient. DESIGN: Interventional case report and literature review. METHODS: A previously well 24-year-old white woman presented with left eye pain and reduced vision. Episcleral injection, globe tenderness, an afferent pupil defect, and exudative retinal detachment were present. Computed tomographic scan of the head and orbits demonstrated scleral thickening, retinal detachment, and no other abnormality. A provisional diagnosis of posterior scleritis with exudative retinal detachment was made. Investigation for underlying connective tissue diseases was negative. There was an initial prompt response to corticosteroid therapy. The patient's symptoms and signs then recurred, and a left third cranial nerve palsy developed. Systemic investigations including lumbar puncture ultimately led to the diagnosis of primary T-cell central nervous system (CNS) lymphoma. Serologic tests for human immunodeficiency virus were negative. MAIN OUTCOME MEASURES AND RESULTS: The patient underwent orbital and cranial irradiation and intrathecal and systemic chemotherapy. Despite an initial response to treatment, she returned with a recurrence of the lymphoma in the anterior segment of the left eye. Her systemic disease progressed rapidly, and she died shortly thereafter. CONCLUSIONS: This patient's young age and initial presentation mimicking posterior scleritis with unilateral exudative retinal detachment, without evidence of vitreous involvement, are highly unusual for ocular involvement in primary CNS lymphoma. A review of the literature highlights the atypical nature of this presentation. PMID- 11013207 TI - Differential electron flow around photosystem I by two C(4)-photosynthetic-cell specific ferredoxins. AB - In the C(4) plant maize (Zea mays L.), two ferredoxin isoproteins, Fd I and Fd II, are expressed specifically in mesophyll and bundle-sheath cells, respectively. cDNAs for these ferredoxins were introduced separately into the cyanobacterium Plectonema boryanum with a disrupted endogenous ferredoxin gene, yielding TM202 and KM2-9 strains expressing Fd I and Fd II. The growth of TM202 was retarded under high light (130 micromol/m(2)/s), whereas KM2-9 grew at a normal rate but exhibited a nitrogen-deficient phenotype. Measurement of photosynthetic O(2) evolution revealed that the reducing power was not efficiently partitioned into nitrogen assimilation in KM2-9. After starvation of the cells in darkness, the P700 oxidation level under far-red illumination increased significantly in TM202. However, it remained low in KM2-9, indicating an active cyclic electron flow. In accordance with this, the cellular ratio of ATP/ADP increased and that of NADPH/NADP(+) decreased in KM2-9 as compared with TM202. These results demonstrated that the two cell type-specific ferredoxins differentially modulate electron flow around photosystem I. PMID- 11013208 TI - A short sequence in the N-terminal region is required for the trimerization of type XIII collagen and is conserved in other collagenous transmembrane proteins. AB - The recombinant transmembrane protein type XIII collagen is shown to reside on the plasma membrane of insect cells in a 'type II' orientation. Expressions of deletion constructs showed that sequences important for the association of three alpha1(XIII) chains reside in their N- rather than C-terminal portion. In particular, a deletion of residues 63-83 immediately adjacent to the transmembrane domain abolished the formation of disulfide-bonded trimers. The results imply that nucleation of the type XIII collagen triple helix occurs at the N-terminal region and that triple helix formation proceeds from the N- to the C-terminus, in opposite orientation to that of the fibrillar collagens. Interestingly, a sequence homologous to the deleted residues was found at the same plasma membrane-adjacent location in other collagenous transmembrane proteins, suggesting that it may be a conserved association domain. The type XIII collagen was secreted into insect cell medium in low amounts, but this secretion was markedly enhanced when the cytosolic portion was lacking. The cleavage occurred in the non-collagenous NC1 domain after four arginines and was inhibited by a furin protease inhibitor. PMID- 11013209 TI - Targeted deletion of keratins 18 and 19 leads to trophoblast fragility and early embryonic lethality. AB - It has been reported previously that keratin 8 (K8)-deficient mice of one strain die from a liver defect at around E12.5, while those of another strain suffer from colorectal hyperplasia. These findings have generated considerable confusion about the function of K8, K18 and K19 that are co-expressed in the mouse blastocyst and internal epithelia. To resolve this issue, we produced mice doubly deficient for K18 and K19 leading to complete loss of keratin filaments in early mouse development. These embryos died at around day E9.5 with 100% penetrance. The absence of keratins caused cytolysis restricted to trophoblast giant cells, followed by haematomas in the trophoblast layer. Up to that stage, embryonic development proceeded unaffected in the absence of keratin filaments. K18/19 deficient mouse embryos die earlier than any other intermediate filament knockouts reported so far, suggesting that keratins, in analogy to their well established role in epidermis, are essential for the integrity of a specialized embryonic epithelium. Our data also offer a rationale to explore the involvement of keratin mutations in early abortions during human pregnancies. PMID- 11013211 TI - Membrane proteins organize a symmetrical virus. AB - Alphaviruses are enveloped icosahedral viruses that mature by budding at the plasma membrane. According to a prevailing model maturation is driven by binding of membrane protein spikes to a preformed nucleocapsid (NC). The T = 4 geometry of the membrane is thought to be imposed by the NC through one-to-one interactions between spike protomers and capsid proteins (CPs). This model is challenged here by a Semliki Forest virus capsid gene mutant. Its CPs cannot assemble into NCs, or its intermediate structures, due to defective CP-CP interactions. Nevertheless, it can use its horizontal spike-spike interactions on membrane surface and vertical spike-CP interactions to make a particle with correct geometry and protein stoichiometry. Thus, our results highlight the direct role of membrane proteins in organizing the icosahedral conformation of alphaviruses. PMID- 11013210 TI - Transfer of palmitate from phospholipids to lipid A in outer membranes of gram negative bacteria. AB - Regulated covalent modifications of lipid A are implicated in virulence of pathogenic Gram-negative bacteria. The Salmonella typhimurium PhoP/PhoQ-activated gene pagP is required both for biosynthesis of hepta-acylated lipid A species containing palmitate and for resistance to cationic anti-microbial peptides. Palmitoylated lipid A can also function as an endotoxin antagonist. We now show that pagP and its Escherichia coli homolog (crcA) encode an unusual enzyme of lipid A biosynthesis localized in the outer membrane. PagP transfers a palmitate residue from the sn-1 position of a phospholipid to the N-linked hydroxymyristate on the proximal unit of lipid A (or its precursors). PagP bearing a C-terminal His(6)-tag accumulated in outer membranes during overproduction, was purified with full activity and was shown by cross-linking to behave as a homodimer. PagP is the first example of an outer membrane enzyme involved in lipid A biosynthesis. Additional pagP homologs are encoded in the genomes of Yersinia and Bordetella species. PagP may provide an adaptive response toward both Mg(2+) limitation and host innate immune defenses. PMID- 11013212 TI - Decreased UDP-GlcNAc levels abrogate proliferation control in EMeg32-deficient cells. AB - The hexosamine pathway provides UDP-N:-acetylhexosamine donor substrates used in cytosolic and Golgi-mediated glycosylation of proteins and for formation of glycosylphosphatidylinositol (GPI) anchors, which tether proteins to the outer plasma membrane. We have recently identified the murine glucosamine-6-phosphate (GlcN6P) acetyltransferase, EMeg32, as a developmentally regulated enzyme on the route to UDP-N:-acetylglucosamine (UDP-GlcNAc). Here we describe embryos and cells that have the EMeg32 gene inactivated by homologous recombination. Homozygous mutant embryos die at around embryonic day (E) 7.5 with a general proliferative delay of development. In vitro differentiated EMeg32(-/-) ES cells show reduced proliferation. Mouse embryonic fibroblasts (MEFs) deficient for EMeg32 exhibit defects in proliferation and adhesiveness, which could be complemented by stable re-expression of EMeg32 or by nutritional restoration of intracellular UDP-GlcNAc levels. Reduced UDP-GlcNAc levels predominantly translated into decreased O-GlcNAc modifications of cytosolic and nuclear proteins. Interestingly, growth-impaired EMeg32(-/-) MEFs withstand a number of apoptotic stimuli and express activated PKB/AKT. Thus, EMeg32-dependent UDP GlcNAc levels influence cell cycle progression and susceptibility to apoptotic stimuli. PMID- 11013213 TI - Crystal structure of the GAP domain of Gyp1p: first insights into interaction with Ypt/Rab proteins. AB - We present the 1.9 A resolution crystal structure of the catalytic domain of Gyp1p, a specific GTPase activating protein (GAP) for Ypt proteins, the yeast homologues of Rab proteins, which are involved in vesicular transport. Gyp1p is a member of a large family of eukaryotic proteins with shared sequence motifs. Previously, no structural information was available for any member of this class of proteins. The GAP domain of Gyp1p was found to be fully alpha-helical. However, the observed fold does not superimpose with other alpha-helical GAPs (e.g. Ras- and Cdc42/Rho-GAP). The conserved and catalytically crucial arginine residue, identified by mutational analysis, is in a comparable position to the arginine finger in the Ras- and Cdc42-GAPs, suggesting that Gyp1p utilizes an arginine finger in the GAP reaction, in analogy to Ras- and Cdc42-GAPs. A model for the interaction between Gyp1p and the Ypt protein satisfying biochemical data is given. PMID- 11013214 TI - p56(dok-2) as a cytokine-inducible inhibitor of cell proliferation and signal transduction. AB - p56(dok-2) acts as a multiple docking protein downstream of receptor or non receptor tyrosine kinases. However, the role of p56(dok-2) in biological functions of cells is not clear. We found that transcription of the p56(dok-2) gene in macrophages was increased markedly in response to cytokines such as macrophage colony-stimulating factor (M-CSF), granulocyte/macrophage-CSF and interleukin-3 (IL-3). Forced expression of p56(dok-2) inhibited M-CSF-, granulocyte-CSF-, IL-3- and stem cell factor-induced proliferation of myeloid leukemia cells, M-NFS-60. The p56(dok-2)-overexpressing cells showed an impaired induction of c-myc but not of c-jun, junB or c-fos when stimulated with M-CSF. Consistent with these results, the peritoneal cavity of the hairless (hr/hr) strain of mutant mice, whose cells expressed less p56(dok-2) than wild-type mice, contained more macrophages than that of +/hr mice. Moreover, the inhibition of endogenous p56(dok-2) expression in macrophage-like tumor cells, J774A.1, by stable expression of antisense p56(dok-2) mRNA accelerated cell proliferation. The study identifies a novel role for p56(dok-2) as a molecule that negatively regulates signal transduction and cell proliferation mediated by cytokines in a feedback loop. PMID- 11013215 TI - CD95 (APO-1/Fas) linkage to the actin cytoskeleton through ezrin in human T lymphocytes: a novel regulatory mechanism of the CD95 apoptotic pathway. AB - CD95 (APO-1/Fas) is a member of the tumor necrosis factor receptor family, which can trigger apoptosis in a variety of cell types. However, little is known of the mechanisms underlying cell susceptibility to CD95-mediated apoptosis. Here we show that human T cells that are susceptible to CD95-mediated apoptosis, exhibit a constitutive polarized morphology, and that CD95 colocalizes with ezrin at the site of cellular polarization. In fact, CD95 co-immunoprecipitates with ezrin exclusively in lymphoblastoid CD4(+) T cells and primary long-term activated T lymphocytes, which are prone to CD95-mediated apoptosis, but not in short-term activated T lymphocytes, which are refractory to the same stimuli, even expressing equal levels of CD95 on the cell membrane. Pre-treatment with ezrin antisense oligonucleotides specifically protected from the CD95-mediated apoptosis. Moreover, we show that the actin cytoskeleton integrity is essential for this function. These findings strongly suggest that the CD95 cell membrane polarization, through an ezrin-mediated association with the actin cytoskeleton, is a key intracellular mechanism in rendering human T lymphocytes susceptible to the CD95-mediated apoptosis. PMID- 11013216 TI - MDM2 induces hyperplasia and premalignant lesions when expressed in the basal layer of the epidermis. AB - The MDM2 oncogene is overexpressed in 5-10% of human tumours. Its major physiological role is to inhibit the tumour suppressor p53. However, MDM2 has p53 independent effects on differentiation and does not predispose to tumorigenesis when it is expressed in the granular layer of the epidermis. These unexpected properties of MDM2 could be tissue specific or could depend on the differentiation state of the cells. Strikingly, we found that MDM2 has p53 dependent effects on differentiation, proliferation and apoptosis when it is expressed in the less differentiated basal layer cells. MDM2 inhibits UV induction of p53, the cell cycle inhibitor p21(WAF1/CIP1) and apoptosis ('sunburn cells'). Importantly, MDM2 increases papilloma formation induced by chemical carcinogenesis and predisposes to the appearance of premalignant lesions and squamous cell carcinomas. p53 has a natural role in the protection against UV damage in the basal layer of the epidermis. Our results show that MDM2 predisposes to tumorigenesis when expressed at an early stage of differentiation, and provide a mouse model of MDM2 tumorigenesis relevant to p53's tumour suppressor functions. PMID- 11013217 TI - A Caenorhabditis elegans MAP kinase kinase, MEK-1, is involved in stress responses. AB - The c-Jun N-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family, was shown to be involved in the response to various stresses in cultured cells. However, there is little in vivo evidence indicating a role for a JNK pathway in the stress response of an organism. We identified the Caenorhabditis elegans mek-1 gene, which encodes a 347 amino acid protein highly homologous to mammalian MKK7, an activator of JNK. Mek-1 reporter fusion proteins are expressed in pharyngeal muscle, uterus, a portion of intestine, and neurons. A mek-1 deletion mutant is hypersensitive to copper and cadmium ions and to starvation. A wild-type mek-1 transgene rescued the hypersensitivity to the metal ions. Double mutants of mek-1 with an eat-5, eat-11 or eat-18 mutation, which are characterized by a limited feeding defect, showed distinct growth defects under normal conditions. Expression of an activated form of MEK-1 in the whole animal or specifically in the pharynx inhibited pharyngeal pumping. These results suggest a role for mek-1 in stress responses, with a focus in the pharynx and/or intestine. PMID- 11013218 TI - H2O2 sensing through oxidation of the Yap1 transcription factor. AB - The yeast transcription factor Yap1 activates expression of antioxidant genes in response to oxidative stress. Yap1 regulation involves nuclear accumulation, but the mechanism sensing the oxidative stress signal remains unknown. We provide biochemical and genetic evidence that upon H2O2 treatment, Yap1 is activated by oxidation and deactivated by enzymatic reduction with Yap1-controlled thioredoxins, thus providing a mechanism for autoregulation. Two cysteines essential for Yap1 oxidation are also essential for its activation by H2O2. The data are consistent with a model in which oxidation of Yap1 leads to disulfide bond formation with the resulting change of conformation masking recognition of the nuclear export signal by Crm1/Xpo1, thereby promoting nuclear accumulation of the protein. In sharp contrast to H2O2, diamide does not lead to the same Yap1 oxidized form and still activates mutants lacking cysteines essential for H2O2 activation, providing a molecular basis for differential activation of Yap1 by these oxidants. This is the first example of an H2O2-sensing mechanism in a eukaryote that exploits the oxidation of cysteines in order to respond rapidly to stress conditions. PMID- 11013219 TI - Crystal structure of FadR, a fatty acid-responsive transcription factor with a novel acyl coenzyme A-binding fold. AB - FadR is a dimeric acyl coenzyme A (acyl CoA)-binding protein and transcription factor that regulates the expression of genes encoding fatty acid biosynthetic and degrading enzymes in Escherichia coli. Here, the 2.0 A crystal structure of full-length FadR is described, determined using multi-wavelength anomalous dispersion. The structure reveals a dimer and a two-domain fold, with DNA-binding and acyl-CoA-binding sites located in an N-terminal and C-terminal domain, respectively. The N-terminal domain contains a winged helix-turn-helix prokaryotic DNA-binding fold. Comparison with known structures and analysis of mutagenesis data delineated the site of interaction with DNA. The C-terminal domain has a novel fold, consisting of a seven-helical bundle with a crossover topology. Careful analysis of the structure, together with mutational and biophysical data, revealed a putative hydrophobic acyl-CoA-binding site, buried in the core of the seven-helical bundle. This structure aids in understanding FadR function at a molecular level, provides the first structural scaffold for the large GntR family of transcription factors, which are keys in the control of metabolism in bacterial pathogens, and could thus be a possible target for novel chemotherapeutic agents. PMID- 11013221 TI - Transcriptional silencing and promoter methylation triggered by double-stranded RNA. AB - Double-stranded RNA induces a post-transcriptional gene silencing process, termed RNAi, in diverse organisms. It is shown here that transcriptional gene silencing accompanied by de novo methylation of a target promoter in plants can be triggered by a double-stranded RNA containing promoter sequences. Similar to the double-stranded RNA involved in RNAi, this promoter double-stranded RNA, which is synthesized in the nucleus, is partially cleaved into small RNAs approximately 23 nucleotides in length. Both transcriptional and post-transcriptional gene silencing can thus be initiated by double-stranded RNAs that enter the same degradation pathway. The results also implicate double-stranded RNA in directing DNA methylation. Different constructs designed to produce double-stranded promoter RNA in various ways were evaluated for their ability to induce gene silencing in tobacco and Arabidopsis. RNA hairpins transcribed from inverted DNA repeats were the most effective trans-acting silencing signals. This strategy could be useful for transcriptionally downregulating genes in a variety of plants. PMID- 11013220 TI - Smad2, Smad3 and Smad4 cooperate with Sp1 to induce p15(Ink4B) transcription in response to TGF-beta. AB - Transforming growth factor-beta (TGF-beta) arrests growth of epithelial cells by inducing the transcription of p15(Ink4B), a cyclin-dependent kinase inhibitor. In this study, we demonstrate that p15(Ink4B) induction was mediated by a TGF-beta induced complex of Smad2, Smad3, Smad4 and Sp1. Mutations in the Sp1- or Smad binding sequences decreased or abolished the TGF-beta responsiveness of the p15(Ink4B) promoter. Interference with, or deficiency in, Smad2, Smad3 or Smad4 functions also reduced or abolished the TGF-beta-dependent p15(Ink4B) induction, whereas the absence of Sp1 reduced the basal and TGF-beta-induced p15(Ink4B) transcription. In the nucleoprotein complex, Smad2 interacted through its C domain with Sp1 and enhanced the DNA binding and transcriptional activity of Sp1. Smad3 interacted indirectly with Sp1 through its association with Smad2 and/or Smad4, and bound directly to the p15(Ink4B) promoter. Finally, Smad4 interacted through its N-domain with Sp1. Our data demonstrate the physical interactions and functional cooperativity of Sp1 with a complex of Smad2, Smad3 and Smad4 in the induction of the p15(Ink4B) gene. These findings explain the tumor suppressor roles of Smad2 and Smad4 in growth arrest signaling by TGF-beta. PMID- 11013222 TI - Targeting the chromatin-remodeling MSL complex of Drosophila to its sites of action on the X chromosome requires both acetyl transferase and ATPase activities. AB - Dosage compensation in Drosophila is mediated by a multiprotein, RNA-containing complex that associates with the X chromosome at multiple sites. We have investigated the role that the enzymatic activities of two complex components, the histone acetyltransferase activity of MOF and the ATPase activity of MLE, may have in the targeting and association of the complex with the X chromosome. Here we report that MLE and MOF activities are necessary for complexes to access the various X chromosome sites. The role that histone H4 acetylation plays in this process is supported by our observations that MOF overexpression leads to the ectopic association of the complex with autosomal sites. PMID- 11013223 TI - Quorum-sensing signal binding results in dimerization of TraR and its release from membranes into the cytoplasm. AB - Promoter binding by TraR and LuxR, the activators of two bacterial quorum-sensing systems, requires their cognate acyl-homoserine lactone (acyl-HSL) signals, but the role the signal plays in activating these transcription factors is not known. Soluble active TraR, when purified from cells grown with the acyl-HSL, contained bound signal and was solely in dimer form. However, genetic and cross-linking studies showed that TraR is almost exclusively in monomer form in cells grown without signal. Adding signal resulted in dimerization of the protein in a concentration-dependent manner. In the absence of signal, monomer TraR localized to the inner membrane while growth with the acyl-HSL resulted in the appearance of dimer TraR in the cytoplasmic compartment. Affinity chromatography indicated that the N-terminus of TraR from cells grown without signal is hidden. Analysis of heterodimers formed between TraR and its deletion mutants localized the dimerization domain to a region between residues 49 and 156. We conclude that binding signal drives dimerization of TraR and its release from membranes into the cytoplasm. PMID- 11013224 TI - Synergistic transcription activation: a dual role for CRP in the activation of an Escherichia coli promoter depending on MalT and CRP. AB - Activation of the Escherichia coli malEp promoter relies on the formation of a higher order structure involving cooperative binding of MalT to promoter-proximal and promoter-distal sites as well as CRP binding to three sites located in between. MalT is the primary activator and one function of CRP is to facilitate cooperative binding of MalT to its cognate sites by bending the intervening DNA. It is shown here that CRP also participates directly in malEp activation. This function is carried out by the molecule of CRP bound to the CRP site centered at 139.5 (CRP site 3). This molecule of CRP recruits RNA polymerase by promoting the binding of the RNA polymerase alpha subunit C-terminal domain (alphaCTD) to DNA immediately downstream from CRP site 3, via a contact between alphaCTD and activating region I of CRP. The action of MalT and CRP at malEp hence provides the example of a novel and complex mechanism for transcriptional synergy in prokaryotes whereby one activator both helps the primary activator to form a productive complex with promoter DNA and interacts directly with RNA polymerase holoenzyme. PMID- 11013225 TI - Mapping the fMet-tRNA(f)(Met) binding site of initiation factor IF2. AB - The interaction between fMet-tRNA(f)(Met) and Bacillus stearothermophilus translation initiation factor IF2 has been characterized. We demonstrate that essentially all thermodynamic determinants governing the stability and the specificity of this interaction are localized within the acceptor hexanucleotide fMet-3'ACCAAC of the initiator tRNA and a fairly small area at the surface of the beta-barrel structure of the 90-amino acid C-terminal domain of IF2 (IF2 C-2). A weak but specific interaction between IF2 C-2 and formyl-methionyl was also demonstrated. The surface of IF2 C-2 interacting with fMet-tRNA(f)(Met) has been mapped using two independent approaches, site- directed mutagenesis and NMR spectroscopy, which yielded consistent results. The binding site comprises C668 and G715 located in a groove accommodating the methionyl side-chain, R700, in the vicinity of the formyl group, Y701 and K702 close to the acyl bond between fMet and tRNA(f)(Met), and the surface lined with residues K702-S660, along which the acceptor arm of the initiator tRNA spans in the direction 3' to 5'. PMID- 11013226 TI - Ribosomal protein L2 is involved in the association of the ribosomal subunits, tRNA binding to A and P sites and peptidyl transfer. AB - Ribosomal proteins L2, L3 and L4, together with the 23S RNA, are the main candidates for catalyzing peptide bond formation on the 50S subunit. That L2 is evolutionarily highly conserved led us to perform a thorough functional analysis with reconstituted 50S particles either lacking L2 or harboring a mutated L2. L2 does not play a dominant role in the assembly of the 50S subunit or in the fixation of the 3'-ends of the tRNAs at the peptidyl-transferase center. However, it is absolutely required for the association of 30S and 50S subunits and is strongly involved in tRNA binding to both A and P sites, possibly at the elbow region of the tRNAs. Furthermore, while the conserved histidyl residue 229 is extremely important for peptidyl-transferase activity, it is apparently not involved in other measured functions. None of the other mutagenized amino acids (H14, D83, S177, D228, H231) showed this strong and exclusive participation in peptide bond formation. These results are used to examine critically the proposed direct involvement of His229 in catalysis of peptide synthesis. PMID- 11013228 TI - Misinsertion and bypass of thymine-thymine dimers by human DNA polymerase iota. AB - Human DNA polymerase iota (pol(iota)) is a recently discovered enzyme that exhibits extremely low fidelity on undamaged DNA templates. Here, we show that poliota is able to facilitate limited translesion replication of a thymine thymine cyclobutane pyrimidine dimer (CPD). More importantly, however, the bypass event is highly erroneous. Gel kinetic assays reveal that pol(iota) misinserts T or G opposite the 3' T of the CPD approximately 1.5 times more frequently than the correct base, A. While pol(iota) is unable to extend the T.T mispair significantly, the G.T mispair is extended and the lesion completely bypassed, with the same efficiency as that of the correctly paired A. T base pair. By comparison, pol(iota) readily misinserts two bases opposite a 6-4 thymine-thymine pyrimidine-pyrimidone photoproduct (6-4PP), but complete lesion bypass is only a fraction of that observed with the CPD. Our data indicate, therefore, that poliota possesses the ability to insert nucleotides opposite UV photoproducts as well as to perform unassisted translesion replication that is likely to be highly mutagenic. PMID- 11013227 TI - Subunit-specific degradation of the UmuD/D' heterodimer by the ClpXP protease: the role of trans recognition in UmuD' stability. AB - The Escherichia coli UmuD' protein is a subunit of the recently described error prone DNA polymerase, pol V. UmuD' is initially synthesized as an unstable and mutagenically inactive pro-protein, UmuD. Upon processing, UmuD' assumes a relatively stable conformation and becomes mutagenically active. While UmuD and UmuD' by themselves exist in vivo as homodimers, when together they preferentially interact to form heterodimers. Quite strikingly, it is in this context that UmuD' becomes susceptible to ClpXP-mediated proteolysis. Here we report a novel targeting mechanism designed for degrading the mutagenically active UmuD' subunit of the UmuD/D' heterodimer complex, while leaving the UmuD protein intact. Surprisingly, a signal that is essential and sufficient for targeting UmuD' for degradation was found to reside on UmuD not UmuD'. UmuD was also shown to be capable of channeling an excess of UmuD' to ClpXP for degradation, thereby providing a mechanism whereby cells can limit error-prone DNA replication. PMID- 11013229 TI - Pituitary tumor transforming gene causes aneuploidy and p53-dependent and p53 independent apoptosis. AB - The pituitary tumor transforming gene, PTTG, is abundantly expressed in several neoplasms. We recently showed that PTTG overexpression is associated with apoptosis and therefore have now studied the role of p53 in this process. In MCF 7 breast cancer cells that express wild type p53, PTTG overexpression caused apoptosis. p53 was translocated to the nuclei in cells expressing PTTG. Overexpression of p53, along with PTTG, augmented apoptosis, whereas expression of the human papillomavirus E6 protein inhibited PTTG-induced apoptosis. In MG-63 osteosarcoma cells that are deficient in p53, PTTG caused cell cycle arrest and subsequent apoptosis that was inhibited by caspase inhibitors. A proteasome inhibitor augmented PTTG expression in stable PTTG transfectants, suggesting that down-regulated PTTG expression is required for cell survival. Finally, MG-63 cells expressing PTTG showed signs of aneuploidy including the presence of micronuclei and multiple nuclei. These results indicate that PTTG overexpression causes p53-dependent and p53-independent apoptosis. In the absence of p53, PTTG causes aneuploidy. These results may provide a mechanism for PTTG-induced tumorigenesis whereby PTTG mediates aneuploidy and subsequent cell transformation. PMID- 11013230 TI - Direct binding of activated c-Src to the beta 3-adrenergic receptor is required for MAP kinase activation. AB - Both beta(2)- and beta(3)-adrenergic receptors (ARs) are able to activate the extracellular signal-regulated kinase (ERK) pathway. We previously showed that c Src is required for ERK activation by beta(2)AR and that it is recruited to activated beta(2)AR through binding of the Src homology 3 (SH3) domain to proline rich regions of the adapter protein beta-arrestin1. Despite the absence of sites for phosphorylation and beta-arrestin binding, ERK activation by beta(3)AR still requires c-Src. Agonist activation of beta(2)AR, but not beta(3)AR, led to redistribution of green fluorescent protein-tagged beta-arrestin to the plasma membrane. In beta-arrestin-deficient COS-7 cells, beta-agonist-dependent co precipitation of c-Src with the beta(2)AR required exogenous beta-arrestin, but activated beta(3)AR co-precipitated c-Src in the absence or presence of beta arrestin. ERK activation and Src co-precipitation with beta(3)AR also occurred in adipocytes in an agonist-dependent and pertussis toxin-sensitive manner. Protein interaction studies show that the beta(3)AR interacts directly with the SH3 domain of Src through proline-rich motifs (PXXP) in the third intracellular loop and the carboxyl terminus. ERK activation and Src co-precipitation were abolished in cells expressing point mutations in these PXXP motifs. Together, these data describe a novel mechanism of ERK activation by a G protein-coupled receptor in which the intracellular domains directly recruit c-Src. PMID- 11013231 TI - Cell wall assembly by Pneumocystis carinii. Evidence for a unique gsc-1 subunit mediating beta -1,3-glucan deposition. AB - Pneumocystis carinii remains a persistent cause of severe pneumonia in immune compromised patients. Recent studies indicate that P. carinii is a fungal species possessing a glucan-rich cyst wall. Pneumocandin antagonists of beta-1,3-glucan synthesis rapidly suppress infection in animal models of P. carinii pneumonia. We, therefore, sought to define the molecular mechanisms of beta-glucan cell wall assembly by P. carinii. Membrane extracts derived from freshly purified P. carinii incorporate uridine 5'-diphosphoglucose into insoluble carbohydrate, in a manner that was completely inhibited by the pneumocandin L733-560, an antagonist of Gsc-1-type beta-glucan synthetases. Using degenerative polymerase chain reaction and library screening, the P. carinii Gsc-1 catalytic subunit of beta 1,3-glucan synthetase was cloned and characterized. P. carinii gsc1 exhibited homology to phylogenetically related fungal beta-1,3-glucan synthetases, encoding a predicted 214-kDa integral membrane protein with 12 transmembrane domain structure. Immunoprecipitation of P. carinii extracts, with a synthetic peptide anti-Gsc-1 antibody, specifically yielded a protein of 219.4 kDa, which was also capable of incorporating 5'-diphosphoglucose into insoluble glucan carbohydrate. As opposed to other fungi, the expression of gsc-1 mRNA is uniquely regulated over P. carinii's life cycle, having minimal expression in trophic forms, but substantial expression in the thick-walled cystic form of the organism. These results indicate that P. carinii contains a unique catalytic subunit of beta-1,3 glucan synthetase utilized in cyst wall formation. Because synthesis of beta-1,3 glucan is absent in mammalian cells, inhibition of the P. carinii Gsc-1 represents an attractive molecular target for therapeutic exploitation. PMID- 11013232 TI - Indirubins inhibit glycogen synthase kinase-3 beta and CDK5/p25, two protein kinases involved in abnormal tau phosphorylation in Alzheimer's disease. A property common to most cyclin-dependent kinase inhibitors? AB - The bis-indole indirubin is an active ingredient of Danggui Longhui Wan, a traditional Chinese medicine recipe used in the treatment of chronic diseases such as leukemias. The antitumoral properties of indirubin appear to correlate with their antimitotic effects. Indirubins were recently described as potent (IC(50): 50-100 nm) inhibitors of cyclin-dependent kinases (CDKs). We report here that indirubins are also powerful inhibitors (IC(50): 5-50 nm) of an evolutionarily related kinase, glycogen synthase kinase-3beta (GSK-3 beta). Testing of a series of indoles and bis-indoles against GSK-3 beta, CDK1/cyclin B, and CDK5/p25 shows that only indirubins inhibit these kinases. The structure activity relationship study also suggests that indirubins bind to GSK-3 beta's ATP binding pocket in a way similar to their binding to CDKs, the details of which were recently revealed by crystallographic analysis. GSK-3 beta, along with CDK5, is responsible for most of the abnormal hyperphosphorylation of the microtubule-binding protein tau observed in Alzheimer's disease. Indirubin-3' monoxime inhibits tau phosphorylation in vitro and in vivo at Alzheimer's disease specific sites. Indirubins may thus have important implications in the study and treatment of neurodegenerative disorders. Indirubin-3'-monoxime also inhibits the in vivo phosphorylation of DARPP-32 by CDK5 on Thr-75, thereby mimicking one of the effects of dopamine in the striatum. Finally, we show that many, but not all, reported CDK inhibitors are powerful inhibitors of GSK-3 beta. To which extent these GSK-3 beta effects of CDK inhibitors actually contribute to their antimitotic and antitumoral properties remains to be determined. Indirubins constitute the first family of low nanomolar inhibitors of GSK-3 beta to be described. PMID- 11013233 TI - Small maf (MafG and MafK) proteins negatively regulate antioxidant response element-mediated expression and antioxidant induction of the NAD(P)H:Quinone oxidoreductase1 gene. AB - The antioxidant response element (ARE) is known to regulate expression and induction of NQO1, GST Ya, and other detoxifying enzyme genes in response to antioxidants and xenobiotics. The nuclear transcription factor Nrf2 and Nrf1 bind to the ARE and positively regulate expression and induction of the NQO1 and GST Ya genes. In this study, we demonstrate that overexpression of small Maf (MafG and MafK) proteins negatively regulate ARE-mediated expression and tert-butyl hydroquinone induction of the NQO1 and GST Ya genes in transfected Hep-G2 cells. In similar experiments, overexpression of small Maf proteins also repressed Nrf2 mediated up-regulation of ARE-mediated NQO1 and GST Ya genes expression in Hep-G2 cells co-transfected with Nrf2 and small Maf proteins. Band and supershift assays with the NQO1 gene ARE and nuclear proteins demonstrate that small MafG and MafK bind to the ARE as Maf-Maf homodimers and Maf-Nrf2 heterodimers. Therefore, Maf Maf homodimers and possibly Maf-Nrf2 heterodimers play a role in negative regulation of ARE-mediated transcription and antioxidant induction of NQO1 and other detoxifying enzyme genes. In contrast to Maf-Nrf2, the Maf-Nrf1 heterodimers failed to bind with the NQO1 gene ARE and did not demonstrate the repressive effect in transfection assays. PMID- 11013234 TI - Identification in Saccharomyces cerevisiae of two isoforms of a novel mitochondrial transporter for 2-oxoadipate and 2-oxoglutarate. AB - The nuclear genome of Saccharomyces cerevisiae encodes 35 members of a family of membrane proteins. Known members transport substrates and products across the inner membranes of mitochondria. We have localized two hitherto unidentified family members, Odc1p and Odc2p, to the inner membranes of mitochondria. They are isoforms with 61% sequence identity, and we have shown in reconstituted liposomes that they transport the oxodicarboxylates 2-oxoadipate and 2-oxoglutarate by a strict counter exchange mechanism. Intraliposomal adipate and glutarate and to a lesser extent malate and citrate supported [14C]oxoglutarate uptake. The expression of Odc1p, the more abundant isoform, made in the presence of nonfermentable carbon sources, is repressed by glucose. The main physiological roles of Odc1p and Odc2p are probably to supply 2-oxoadipate and 2-oxoglutarate from the mitochondrial matrix to the cytosol where they are used in the biosynthesis of lysine and glutamate, respectively, and in lysine catabolism. PMID- 11013235 TI - Characterization of an upstream enhancer region in the promoter of the human endothelial nitric-oxide synthase gene. AB - The endothelial nitric-oxide synthase gene is constitutively expressed in endothelial cells. Several transcriptionally active regulatory elements have been identified in the proximal promoter, including a GATA-2 and an Sp-1 binding site. Because they cannot account for the constitutive expression of endothelial nitric oxide synthase gene in a restricted number of cells, we have searched for other cell-specific regulatory elements. By DNase I hypersensitivity mapping and deletion studies we have identified a 269-base pair activator element located 4.9 kilobases upstream from the transcription start site that acts as an enhancer. DNase I footprinting and linker-scanning experiments showed that several regions within the 269-base pair enhancer are important for transcription factor binding and for full enhancer activity. The endothelial specificity of this activation seems partly due to interaction between this enhancer in its native configuration and the promoter in endothelial cells. EMSA experiments suggested the implication of MZF-like, AP-2, Sp-1-related, and Ets-related factors. Among Ets factors, Erg was the only one able to bind to cognate sites in the enhancer, as found by EMSA and supershift experiments, and to activate the transcriptional activity of the enhancer in cotransfection experiments. Therefore, multiple protein complexes involving Erg, other Ets-related factors, AP-2, Sp-1-related factor, and MZF-like factors are important for the function of this enhancer in endothelial cells. PMID- 11013236 TI - Intron-exon swapping of transglutaminase mRNA and neuronal Tau aggregation in Alzheimer's disease. AB - In order to understand the mechanism for insoluble neurotoxic protein polymerization in Alzheimer's disease (AD) brain neurons, we examined protein and gene expression for transglutaminase (TGase 2; tissue transglutaminase (tTG)) in hippocampus and isocortex. We found co-localization of tTG protein and activity with tau-positive neurofibrillary tangles, whereas mRNA and sequence analysis indicated an absolute increase in tTG synthesized. Although apoptosis in AD hippocampus is now an established mode of neuronal cell death, no definite underlying mechanism(s) is known. Since TGase-mediated protein aggregation is implicated in polyglutamine ((CAG)(n)/Q(n) expansion) disorder apoptosis, and expanded Q(n) repeats are excellent TGase substrates, a role for TGase in AD is possible. However, despite such suggestions almost 20 years ago, the molecular mechanism remained elusive. We now present one possible molecular mechanism for tTG-mediated, neurotoxic protein polymerization leading to neuronal apoptosis in AD that involves not its substrates (like Q(n) repeats) but rather the unique presence of alternative transcripts of tTG mRNA. In addition to a full-length (L) isoform in aged non-demented brains, we found a short isoform (S) lacking a binding domain in all AD brains. Our current results identify intron-exon "switching" between L and S isoforms, implicating G-protein-coupled signaling pathways associated with tTG that may help to determine the dual roles of this enzyme in neuronal life and death processes. PMID- 11013237 TI - Regulation of glycogen synthesis by amino acids in cultured human muscle cells. AB - Insulin and a number of metabolic factors stimulate glycogen synthesis and the enzyme glycogen synthase. Using human muscle cells we find that glycogen synthesis is stimulated by treatment of the cells with lithium ions, which inhibit glycogen synthase kinase 3. Insulin further stimulates glycogen synthesis in the presence of lithium ions, an effect abolished by wortmannin and rapamycin. We report also that amino acids stimulate glycogen synthesis and glycogen synthase, these effects also being blocked by rapamycin and wortmannin. Amino acids stimulate p70(s6k) and transiently inhibit glycogen synthase kinase 3 without effects on the activity of protein kinase B or the mitogen-activated protein kinase pathway. Thus, the work reported here demonstrates that amino acid availability can regulate glycogen synthesis. Furthermore, it demonstrates that glycogen synthase kinase 3 can be inactivated within cells independent of activation of protein kinase B and p90(rsk). PMID- 11013238 TI - Eosinophil peroxidase oxidation of thiocyanate. Characterization of major reaction products and a potential sulfhydryl-targeted cytotoxicity system. AB - Although the pseudohalide thiocyanate (SCN(-)) is the preferred substrate for eosinophil peroxidase (EPO) in fluids of physiologic halide composition, the product(s) of this reaction have not been directly identified, and mechanisms underlying their cytotoxic potential are poorly characterized. We used nuclear magnetic resonance spectroscopy (NMR), electrospray ionization mass spectrometry, and quantitative chemical analysis to identify the principal reaction products of both the EPO/SCN(-)/H(2)O(2) system and activated eosinophils as roughly equimolar amounts of OSCN(-) (hypothiocyanite) and OCN(-) (cyanate). Red blood cells exposed to increasing concentrations of OSCN(-)/OCN(-) are first depleted of glutathione, after which glutathione S-transferase and glyceraldehyde-3 phosphate dehydrogenase then ATPases undergo sulfhydryl (SH) reductant-reversible inactivation before lysing. OSCN(-)/OCN(-) inactivates red blood cell membrane ATPases 10-1000 times more potently than do HOCl, HOBr, and H(2)O(2). Exposure of glutathione S-transferase to [(14)C]OSCN(-)/OCN(-) causes SH reductant-reversible disulfide bonding and covalent isotope labeling. We propose that EPO/SCN( )/H(2)O(2) reaction products comprise a potential SH-targeted cytotoxic system that functions in striking contrast to HOCl, the highly but relatively indiscriminantly reactive product of the neutrophil myeloperoxidase system. PMID- 11013239 TI - Specific desensitization of glycogen synthase activation by insulin in 3T3-L1 adipocytes. Connection between enzymatic activation and subcellular localization. AB - A protocol was developed in 3T3-L1 adipocytes that resulted in the specific desensitization of glycogen synthase activation by insulin. Cells were pretreated for 15 min with 100 nm insulin, and then recovered for 1.5 h in the absence of hormone. Subsequent basal and insulin-induced phosphorylation of the insulin receptor, IRS-1, MAPK, Akt kinase, and GSK-3 were similar in control and pretreated cells. Additionally, enhanced glucose transport and incorporation into lipid in response to insulin were unaffected. However, pretreatment reduced insulin-stimulated glycogen synthesis by over 50%, due to a nearly complete inhibition of glycogen synthase activation. Removal of extracellular glucose during the recovery period blocked the increase in glycogen levels, and restored insulin-induced glycogen synthase activation. Furthermore, incubation of pretreated 3T3-L1 adipocytes with glycogenolytic agents reversed the desensitization event. Separation of cellular lysates on sucrose gradients revealed that glycogen synthase was primarily located in the dense pellet fraction, with lesser amounts in the lighter fractions. Insulin induced glycogen synthase translocation from the lighter to the denser glycogen-containing fractions. Interestingly, insulin preferentially activated translocated enzyme while having little effect on the majority of glycogen synthase activity in the pellet fraction. In insulin-pretreated cells, glycogen synthase did not return to the lighter fractions during recovery, and thus did not move in response to the second insulin exposure. These results suggest that, in 3T3-L1 adipocytes, the translocation of glycogen synthase may be an important step in the regulation of glycogen synthesis by insulin. Furthermore, intracellular glycogen levels can regulate glycogen synthase activation, potentially through modulation of enzymatic localization. PMID- 11013240 TI - A loss of function mutant of the presenilin homologue SEL-12 undergoes aberrant endoproteolysis in Caenorhabditis elegans and increases abeta 42 generation in human cells. AB - The familial Alzheimer's disease-associated presenilins (PSs) occur as a dimeric complex of proteolytically generated fragments, which functionally supports endoproteolysis of Notch and the beta-amyloid precursor protein (betaAPP). A homologous gene, sel-12, has been identified in Caenorhabditis elegans. We now demonstrate that wild-type (wt) SEL-12 undergoes endoproteolytic cleavage in C. elegans similar to the PSs in human tissue. In contrast, SEL-12 C60S protein expressed from the sel-12(ar131) allele is miscleaved in C. elegans, resulting in a larger mutant N-terminal fragment. Neither SEL-12 wt nor C60S undergo endoproteolytic processing upon expression in human cells, suggesting that SEL-12 is cleaved by a C. elegans-specific endoproteolytic activity. The loss of function of sel-12 in C. elegans is not associated with a dominant negative activity in human cells, because SEL-12 C60S and the corresponding PS1 C92S mutation do not interfere with Notch1 cleavage. Moreover, both mutant variants increase the aberrant production of the highly amyloidogenic 42-amino acid version of amyloid beta-peptide similar to familial Alzheimer's disease associated human PS mutants. Our data therefore demonstrate that the C60S mutation in SEL-12 is associated with aberrant endoproteolysis and a loss of function in C. elegans, whereas a gain of misfunction is observed upon expression in human cells. PMID- 11013241 TI - Thyroglobulin is selected as luminal protein cargo for apical transport via detergent-resistant membranes in epithelial cells. AB - Thyroid hormone synthesis by thyrocytes depends upon apical secretion of thyroglobulin (Tg), the glycoprotein prohormone. In stably transfected MDCK cells, recombinant Tg is also secreted apically. All secreted Tg has undergone Golgi carbohydrate modification, whereas most intracellular Tg (which is slow to exit the endoplasmic reticulum) is sensitive to digestion with endoglycosidase H. However, in MDCK cells and PC Cl3 thyrocytes, a subpopulation of newly synthesized recombinant and endogenous Tg, respectively, is recovered in a Triton X-100 insoluble, glycosphingolipid/cholesterol-enriched (GEM/raft) fraction, and this small subpopulation is overwhelmingly endoglycosidase H resistant. Upon apical secretion, Tg solubility is restored. Apical secretion of Tg is inhibited by cellular cholesterol depletion. In FRT cells, recombinant Tg becomes Triton X 100 insoluble within 60 min after synthesis and a portion is actually endoglycosidase H-sensitive, suggesting early Tg entry into GEMs/rafts. Interestingly in FRT cells, Tg remains associated with the apical plasma membrane upon exocytosis, and all surface Tg is GEM/raft-associated. Thus, Tg is the first secretory protein demonstrated to enter Triton X-100 insoluble membranes en route to the apical surface of epithelial cells. The data imply that Tg utilizes a cargo-selective mechanism for apical sorting. PMID- 11013242 TI - Reverse methionine biosynthesis from S-adenosylmethionine in eukaryotic cells. AB - The intracellular ratio between methionine and its activated form S adenosylmethionine (AdoMet) is of crucial importance for the one-carbon metabolism. AdoMet recycling into methionine was believed to be largely achieved through the methyl and the thiomethyladenosine cycles. We show here that in yeast, AdoMet recycling actually occurs mainly through the direct AdoMet dependent remethylation of homocysteine. Compelling evidences supporting this result were obtained owing to the identification and functional characterization of two new genes, SAM4 and MHT1, that encode the yeast AdoMet-homocysteine methyltransferase and S-methylmethionine-homocysteine methyltransferase, respectively. Homologs of the Sam4 and Mht1 proteins exist in other eucaryotes, indicating that such enzymes would be universal and not restricted to the bacterial or fungal kingdoms. New pathways for AdoMet or S-methylmethionine dependent methionine synthesis are presented. PMID- 11013243 TI - Tyrosine 220 in the 5th transmembrane domain of the neuromedin B receptor is critical for the high selectivity of the peptoid antagonist PD168368. AB - Peptoid antagonists are increasingly being described for G protein-coupled receptors; however, little is known about the molecular basis of their binding. Recently, the peptoid PD168368 was found to be a potent selective neuromedin B receptor (NMBR) antagonist. To investigate the molecular basis for its selectivity for the NMBR over the closely related receptor for gastrin-releasing peptide (GRPR), we used a chimeric receptor approach and a site-directed mutagenesis approach. Mutated receptors were transiently expressed in Balb 3T3. The extracellular domains of the NMBR were not important for the selectivity of PD168368. However, substitution of the 5th upper transmembrane domain (uTM5) of the NMBR by the comparable GRPR domains decreased the affinity 16-fold. When the reverse study was performed by substituting the uTM5 of NMBR into the GRPR, a 9 fold increase in affinity occurred. Each of the 4 amino acids that differed between NMBR and GRPR in the uTM5 region were exchanged, but only the substitution of Phe(220) for Tyr in the NMBR caused a decrease in affinity. When the reverse study was performed to attempt to demonstrate a gain of affinity in the GRPR, the substitution of Tyr(219) for Phe caused an increase in affinity. These results suggest that the hydroxyl group of Tyr(220) in uTM5 of NMBR plays a critical role for high selectivity of PD168368 for NMBR over GRPR. Receptor and ligand modeling suggests that the hydroxyl of the Tyr(220) interacts with nitrophenyl group of PD168368 likely primarily by hydrogen bonding. This result shows the selectivity of the peptoid PD168368, similar to that reported for numerous non-peptide analogues with other G protein-coupled receptors, is primarily dependent on interaction with transmembrane amino acids. PMID- 11013244 TI - Hepatocyte growth factor activator inhibitor type 1 is a specific cell surface binding protein of hepatocyte growth factor activator (HGFA) and regulates HGFA activity in the pericellular microenvironment. AB - Hepatocyte growth factor activator (HGFA) is responsible for proteolytic activation of the precursor form of hepatocyte growth factor in injured tissues. To date, two specific inhibitors of HGFA have been identified, namely HGFA inhibitor type 1 (HAI-1) and type 2 (HAI-2)/placental bikunin (PB). Both inhibitors are first synthesized as integral membrane proteins having two Kunitz domains and a transmembrane domain, and are subsequently released from cell surface by shedding. Here we show that an active form of HGFA is specifically complexed with membrane-form HAI-1, but not with HAI-2/PB, on the surface of epithelial cells expressing both inhibitors. This binding required the enzyme activity of HGFA. The selective binding of HGFA to the cell surface HAI-1 was further confirmed in an engineered system using Chinese hamster ovary cells, in which only the cells expressing HAI-1 retained exogenous HGFA. The binding of HGFA to HAI-1 was reversible, and no irreversible modifications affecting the enzyme activity occurred during the binding. Importantly, HAI-1 and the HGFA.HAI 1 complex were quickly released from the cell surface by treatment with phorbol 12-myristate 13-acetate or interleukin 1beta accompanying the generation of 58 kDa fragments of HAI-1, which are less potent against HGFA, as well as significant recovery of HGFA activity in the culture supernatant. This regulated shedding was completely inhibited by BB3103, a synthetic zinc-metalloproteinase inhibitor. We conclude that HAI-1 is not only an inhibitor but also a specific acceptor of active HGFA, acting as a reservoir of this enzyme on the cell surface. The latter property appears to ensure the concentrated pericellular HGFA activity in certain cellular conditions, such as tissue injury and inflammation, via the up-regulated shedding of HGFA.HAI-1 complex. These findings shed light on a novel function of the integral membrane Kunitz-type inhibitor in the regulation of pericellular proteinase activity. PMID- 11013245 TI - p13(SUC1) and the WW domain of PIN1 bind to the same phosphothreonine-proline epitope. AB - The WW domain of the human PIN1 and p13(SUC1), a subunit of the cyclin-dependent kinase complex, were previously shown to be involved in the regulation of the cyclin-dependent kinase complex activity at the entry into mitosis, by an unresolved molecular mechanism. We report here experimental evidence for the direct interaction of p13(SUC1) with a model CDC25 peptide, dependent on the phosphorylation state of its threonine. Chemical shift perturbation of backbone (1)H(N), (15)N, and (13)Calpha resonances during NMR titration experiments allows accurate identification of the binding site, primarily localized around the anion binding site, occupied in the crystal structure of the homologous p9(CKSHs2) by a sulfate molecule. The epitope recognized by p13(SUC1) includes the proline at position +1 of the phosphothreonine, as was shown by the decrease in affinity for a mutated CDC25 phosphopeptide, containing an alanine/proline substitution. No direct interaction between the PIN1 WW domain or its catalytic proline cis/trans isomerase domain and p13(SUC1) was detected, but our study showed that in vitro the WW domain of the human PIN1 antagonizes the binding of the p13(SUC1) to the CDC25 phosphopeptide, by binding to the same phosphoepitope. We thus propose that the full cyclin-dependent kinase complex stimulates the phosphorylation of CDC25 through binding of its p13(SUC1) module to the phosphoepitope of the substrate and that the reported WW antagonism of p13(SUC1)-stimulated CDC25 phosphorylation is caused by competitive binding of both protein modules to the same phosphoepitope. PMID- 11013246 TI - Tau filament formation in transgenic mice expressing P301L tau. AB - Mutations in the microtubule-associated protein tau, including P301L, are genetically coupled to hereditary frontotemporal dementia with parkinsonism linked to chromosome 17. To determine whether P301L is associated with fibril formation in mice, we expressed the longest human tau isoform, human tau40, with this mutation in transgenic mice by using the neuron-specific mouse Thy1.2 promoter. We obtained mice with high expression of human P301L tau in cortical and hippocampal neurons. Accumulated tau was hyperphosphorylated and translocated from axonal to somatodendritic compartments and was accompanied by astrocytosis and neuronal apoptosis indicated by terminal deoxynucleotidyl transferase mediated biotinylated dUTP nick end-labeling staining. Moreover, P301L tau formed abnormal filaments. Electron microscopy of sarcosyl-insoluble protein extracts established that the filaments had a straight or twisted structure of variable length and were approximately 15 nm wide. Immunoelcecton microscopy showed that the tau filaments were phosphorylated at the TG3, AT100, AT8, and AD199 epitopes in vivo. In cortex, brain stem, and spinal cord, neurofibrillary tangles were also identified by thioflavin-S fluorescent microscopy and Gallyas silver stains. Together, our results show that expression of the P301L mutation in mice causes neuronal lesions that are similar to those seen in human tauopathies. PMID- 11013247 TI - Calmodulin kinase II attenuation of gene transcription by preventing cAMP response element-binding protein (CREB) dimerization and binding of the CREB binding protein. AB - Calmodulin Kinase II (CamKII) inhibits the transcription of many CRE-dependent genes, but the mechanism of dominant transcriptional inhibition is unknown. Here we show that phosphorylation of serine 142 in CREB by CamKII leads to dissociation of the CREB dimer without impeding DNA binding capacity. CamKII modified CREB binds to DNA efficiently as a monomer; however, monomeric CREB is unable to recruit the CREB-binding protein (CBP) even when phosphorylated at serine 133. Thus, CamKII confers a dominant inhibitory effect on transcription by preventing dimerization of CREB, and this mechanism may account for the attenuation of gene expression. PMID- 11013248 TI - Intrinsically bent DNA in the promoter regions of the yeast GAAL1-10 and GAL80 genes. AB - Circular permutation analysis has detected fairly strong sites of intrinsic DNA bending on the promoter regions of the yeast GAL1-10 and GAL80 genes. These bends lie in functionally suggestive locations. On the promoter of the GAL1-10 structural genes, strong bends bracket nucleosome B, which lies between the UAS(G) and the GAL1 TATA. These intrinsic bends could help position nucleosome B. Nucleosome B plus two other promoter nucleosomes protect the TATA and start site elements in the inactive state of expression but are completely disrupted (removed) when GAL1-10 expression is induced. The strongest intrinsic bend ( approximately 70 degrees ) lies at the downstream edge of nucleosome B; this places it approximately 30 base pairs upstream of the GAL1 TATA, a position that could allow it to be involved in GAL1 activation in several ways, including the recruitment of a yeast HMG protein that is required for the normally robust level of GAL1 expression in the induced state (Paull, T., Carey, M., and Johnson, R. (1996) Genes Dev. 10, 2769-2781). On the regulatory gene GAL80, the single bend lies in the non-nucleosomal hypersensitive region, between a GAL80-specific far upstream promoter element and the more gene-proximal promoter elements. GAL80 promoter region nucleosomes contain no intrinsically bent DNA. PMID- 11013249 TI - A single arginyl residue in plastocyanin and in cytochrome c(6) from the cyanobacterium Anabaena sp. PCC 7119 is required for efficient reduction of photosystem I. AB - Positively charged plastocyanin from Anabaena sp. PCC 7119 was investigated by site-directed mutagenesis. The reactivity of its mutants toward photosystem I was analyzed by laser flash spectroscopy. Replacement of arginine at position 88, which is adjacent to the copper ligand His-87, by glutamine and, in particular, by glutamate makes plastocyanin reduce its availability for transferring electrons to photosystem I. Such a residue in the copper protein thus appears to be isofunctional with Arg-64 (which is close to the heme group) in cytochrome c(6) from Anabaena (Molina-Heredia, F. P., Diaz-Quintana, A., Hervas, M., Navarro, J. A., and De la Rosa, M. A. (1999) J. Biol. Chem. 274, 33565-33570) and Synechocystis (De la Cerda, B., Diaz-Quintana, A., Navarro, J. A. , Hervas, M., and De la Rosa, M. A. (1999) J. Biol. Chem. 274, 13292-13297). Other mutations concern specific residues of plastocyanin either at its positively charged east face (D49K, H57A, H57E, K58A, K58E, Y83A, and Y83F) or at its north hydrophobic pole (L12A, K33A, and K33E). Mutations altering the surface electrostatic potential distribution allow the copper protein to modulate its kinetic efficiency: the more positively charged the interaction site, the higher the rate constant. Whereas replacement of Tyr-83 by either alanine or phenylalanine has no effect on the kinetics of photosystem I reduction, Leu-12 and Lys-33 are essential for the reactivity of plastocyanin. PMID- 11013250 TI - Evidence that silencing of the HPRT promoter by DNA methylation is mediated by critical CpG sites. AB - The strong correlation between promoter hypermethylation and gene silencing suggests that promoter methylation represses transcription. To identify methylation sites that may be critical for maintaining repression of the human HPRT gene, we treated human/hamster hybrid cells containing an inactive human X chromosome with the DNA demethylating agent 5-azadeoxycytidine (5aCdr), and we then examined the high resolution methylation pattern of the HPRT promoter in single cell-derived lines. Reactivation of HPRT correlated with complete promoter demethylation. In contrast, the 61 5aCdr-treated clones that failed to reactivate HPRT exhibited sporadic promoter demethylation. However, three specific CpG sites remained methylated in all unreactivated clones, suggesting these sites may be critical for maintaining transcriptional silencing of the HPRT gene. Re-treatment of partially demethylated (and unreactivated) clones with a second round of 5aCdr did not increase the frequency of HPRT reactivation. This is consistent with mechanisms of methylation-mediated repression requiring methylation at specific critical sites and argues against models invoking overall levels or a threshold of promoter methylation. Treatment of cells with the histone deacetylase inhibitor, trichostatin A, failed to reactivate HPRT on the inactive X chromosome, even when the promoter was partially demethylated by 5aCdr treatment, suggesting that transcriptional repression by DNA methylation is unlikely to depend upon a trichostatin A-sensitive histone deacetylase. PMID- 11013251 TI - High and low levels of cottontail rabbit papillomavirus E2 protein generate opposite effects on gene expression. AB - The papillomavirus E2 protein plays an important role in viral transcriptional regulation and replication. We chose to study the cottontail rabbit papillomavirus (CRPV) E2 protein as a transcriptional regulator because of the availability of an animal model for papilloma formation, which may be relevant for human papillomavirus (HPV) infection and replication. We studied the effect of expression levels of E2 on the long control region, which contains transcriptional promoter and enhancer elements, and synthetic E2-dependent artificial promoters in which the E2 was the dominant factor in the transcriptional activation. These experiments indicated that high levels of E2 were inhibitory and low levels were stimulatory for transactivation. In addition, we showed that the complex formed between CRPV E2 and the cognate binding site was less stable than the complex formed between HPV E2 and the same cognate binding site. Furthermore, we showed that CRPV E2 binding to its transcriptional regulatory sequence was stabilized by other proteins such as E1, which produced increments in transcriptional activation of E2-dependent genes. The data may be used to define conditions in which the rabbit model can be used for the screening of drugs which are inhibitory to the HPV and CRPV replication and gene expression. PMID- 11013253 TI - The gene encoding p202, an interferon-inducible negative regulator of the p53 tumor suppressor, is a target of p53-mediated transcriptional repression. AB - The p53 tumor suppressor protein regulates the transcription of regulatory genes involved in cell cycle arrest and apoptosis. We reported previously that overexpression of p202, an interferon-inducible negative regulator of cell growth, negatively regulates the transcriptional activity of p53. Now we identify the gene encoding p202 as one whose mRNA and protein expression decrease in cells following the expression of wild-type, but not mutant, p53. Furthermore, the levels of p202 also decrease after exposure of cells to ultra violet light, which correlate with increase in the levels of p53. We report that the sequence specific DNA binding of p53 to the 5'-regulatory region of the 202 gene contributes to the transcriptional repression of the 202 gene. Interestingly, overexpression of p202 in cells induced to undergo p53-dependent apoptosis significantly delays this process, indicating that the negative regulation of the 202 gene by wild-type p53 is important to potentiate apoptosis. PMID- 11013254 TI - Aldehyde dehydrogenase 6, a cytosolic retinaldehyde dehydrogenase prominently expressed in sensory neuroepithelia during development. AB - We have isolated the chick and mouse homologs of human aldehyde dehydrogenase 6 (ALDH6) that encode a third cytosolic retinaldehyde-specific aldehyde dehydrogenase. In both chick and mouse embryos, strong expression is observed in the sensory neuroepithelia of the head. In situ hybridization analysis in chick shows compartmentalized expression primarily in the ventral retina, olfactory epithelium, and otic vesicle; additional sites of expression include the isthmus, Rathke's pouch, posterior spinal cord interneurons, and developing limbs. Recombinant chick ALDH6 has a K(0.5) = 0.26 microm, V(max) = 48.4 nmol/min/mg and exhibits strong positive cooperativity (H = 1.9) toward all-trans-retinaldehyde; mouse ALDH6 has similar kinetic parameters. Expression constructs can confer 1000 fold increased sensitivity to retinoic acid receptor-dependent signaling from retinol in transient transfections experiments. The localization of ALDH6 to the developing sensory neuroepithelia of the eye, nose, and ear and discreet sites within the CNS suggests a role for RA signaling during primary neurogenesis at these sites. PMID- 11013255 TI - Cloning and characterization of the human activity-dependent neuroprotective protein. AB - We have recently cloned the mouse activity-dependent neuroprotective protein (ADNP). Here, we disclose the cloning of human ADNP (hADNP) from a fetal brain cDNA library. Comparative sequence analysis of these two ADNP orthologs indicated 90% identity at the mRNA level. Several single nucleotide polymorphic sites were noticed. The deduced protein structure contained nine zinc fingers, a proline rich region, a nuclear bipartite localization signal, and a homeobox domain profile, suggesting a transcription factor function. Further comparative analysis identified an ADNP paralog (33% identity and 46% similarity), indicating that these genes belong to a novel protein family with a nine-zinc finger motif followed by a homeobox domain. The hADNP gene structure spans approximately 40 kilobases and includes five exons and four introns with alternative splicing of an untranslated second exon. The hADNP gene was mapped to chromosome 20q12-13.2, a region associated with aggressive tumor growth, frequently amplified in many neoplasias, including breast, bladder, ovarian, pancreatic, and colon cancers. hADNP mRNA is abundantly expressed in distinct normal tissues, and high expression levels were encountered in malignant cells. Down-regulation of ADNP by antisense oligodeoxynucleotides up-regulated the tumor suppressor p53 and reduced the viability of intestinal cancer cells by 90%. Thus, ADNP is implicated in maintaining cell survival, perhaps through modulation of p53. PMID- 11013256 TI - The loop region covering the iron-sulfur cluster in bovine adrenodoxin comprises a new interaction site for redox partners. AB - The amino acid in position 49 in bovine adrenodoxin is conserved among vertebrate [2Fe-2S] ferredoxins as hydroxyl function. A corresponding residue is missing in the cluster-coordinating loop of plant-type [2Fe-2S] ferredoxins. To probe the function of Thr-49 in a vertebrate ferredoxin, replacement mutants T49A, T49S, T49L, and T49Y, and a deletion mutant, T49Delta, were generated and expressed in Escherichia coli. CD spectra of purified proteins indicate changes of the [2Fe 2S] center geometry only for mutant T49Delta, whereas NMR studies reveal no transduction of structural changes to the interaction domain. The redox potential of T49Delta (-370 mV) is lowered by approximately 100 mV compared with wild type adrenodoxin and reaches the potential range of plant-type ferredoxins (-305 to 455 mV). Substitution mutants show moderate changes in the binding affinity to the redox partners. In contrast, the binding affinity of T49Delta to adrenodoxin reductase and cytochrome P-450 11A1 (CYP11A1) is dramatically reduced. These results led to the conclusion that Thr-49 modulates the redox potential in adrenodoxin and that the cluster-binding loop around Thr-49 represents a new interaction region with the redox partners adrenodoxin reductase and CYP11A1. In addition, variations of the apparent rate constants of all mutants for CYP11A1 reduction indicate the participation of residue 49 in the electron transfer pathway between adrenodoxin and CYP11A1. PMID- 11013257 TI - The thioredoxin system of the malaria parasite Plasmodium falciparum. Glutathione reduction revisited. AB - In most living cells, redox homeostasis is based both on the glutathione and the thioredoxin system. In the malaria parasite Plasmodium falciparum antioxidative proteins represent promising targets for the development of antiparasitic drugs. We cloned and expressed a thioredoxin of P. falciparum (pftrx), and we improved the stable expression of the thioredoxin reductase (PfTrxR) of the parasite by multiple silent mutagenesis. Both proteins were biochemically characterized and compared with the human host thioredoxin system. Intriguingly, the 13-kDa protein PfTrx is a better substrate for human TrxR (K(m) = 2 microm, k(cat) = 3300 min( )(1)) than for P. falciparum TrxR (K(m) = 10.4 microm, k(cat) = 3100 min(-)(1)). Possessing a midpoint potential of -270 mV, PfTrx was found to reduce the disease related metabolites S-nitrosoglutathione and GSSG. The rate constant k(2) for the reaction between reduced P. falciparum thioredoxin and GSSG was determined to be 0.039 microm(-)(1) min(-)(1) at 25 degrees C and pH 7.4. The k(2) for thioredoxins from man, Drosophila melanogaster, and Escherichia coli was approximately 5 times lower. Our data suggest that GSSG reduction can be supported at a high rate by the TrxR/Trx system in glutathione reductase deficient cells; this may be relevant for certain stages of the malarial parasite but also for cells containing high [GSSG] of other organisms like dormant forms of Neurospora, glutathione reductase-deficient yeast mutants, or CD4(+) lymphocytes of AIDS patients. PMID- 11013258 TI - Two basic residues of the h-VPAC1 receptor second transmembrane helix are essential for ligand binding and signal transduction. AB - We mutated the vasoactive intestinal peptide (VIP) Asp(3) residue and two VPAC(1) receptor second transmembrane helix basic residues (Arg(188) and Lys(195)). VIP had a lower affinity for R188Q, R188L, K195Q, and K195I VPAC(1) receptors than for VPAC(1) receptors. [Asn(3)] VIP and [Gln(3)] VIP had lower affinities than VIP for VPAC(1) receptors but higher affinities for the mutant receptors; the two basic amino acids facilitated the introduction of the negatively charged aspartate inside the transmembrane domain. The resulting interaction was necessary for receptor activation. 1/[Asn(3)] VIP and [Gln(3)] VIP were partial agonists at VPAC(1) receptors; 2/VIP did not fully activate the K195Q, K195I, R188Q, and R188L VPAC(1) receptors; a VIP analogue ([Arg(16)] VIP) was more efficient than VIP at the four mutated receptors; and [Asn(3)] VIP and [Gln(3)] VIP were more efficient than VIP at the R188Q and R188L VPAC(1) receptors; 3/the [Asp(3)] negative charge did not contribute to the recognition of the VIP(1) antagonist, [AcHis(1),D-Phe(2),Lys(15),Arg(16),Leu(27)] VIP ()/growth hormone releasing factor (8-27). This is the first demonstration that, to activate the VPAC(1) receptor, the Asp(3) side chain of VIP must penetrate within the transmembrane domain, in close proximity to two highly conserved basic amino acids from transmembrane 2. PMID- 11013260 TI - Transmembrane topology of the secretory Na+-K+-2Cl- cotransporter NKCC1 studied by in vitro translation. AB - The secretory Na(+)-K(+)-2Cl(-) cotransporter NKCC1 is a member of a small gene family of electroneutral salt transporters. Hydropathy analyses indicate that all of these transporters have a similar general structure consisting of large hydrophilic N and C termini on either side of a central, relatively well conserved, hydrophobic domain. Programs that predict the transmembrane topology of polytopic membrane proteins identify 10-12 putative membrane-spanning segments (MSSs) in this hydrophobic domain; but to date, there is little experimental data on the structure of this region for any of these transporters. In this report, we have studied the transmembrane topology of NKCC1 using an in vitro translation system designed to test the membrane insertion properties of putative MSSs (Bamberg, K., and Sachs, G. (1994) J. Biol. Chem. 269, 16909-16919). Fusion proteins consisting of putative NKCC1 MSSs inserted either (i) between an N terminal cytosolic anchor sequence and a C-terminal reporter sequence containing multiple N-linked glycosidation sites or (ii) between an N-terminal signal anchor sequence and the same glycosidation flag were expressed in the presence of canine pancreatic microsomes. The glycosidation status of the reporter sequence, which indicated its luminal or extraluminal location in the microsomes, was then used to characterize the signal anchor or stop transfer activity of the inserted MSSs. The results of this experimental analysis yielded a topology scheme consisting of 12 membrane-spanning segments, two pairs of which apparently form rather tight hairpin-like structures within the membrane. PMID- 11013259 TI - Identification of residues within the drug-binding domain of the human multidrug resistance P-glycoprotein by cysteine-scanning mutagenesis and reaction with dibromobimane. AB - P-glycoprotein (P-gp) can transport a wide variety of cytotoxic compounds that have diverse structures. Therefore, the drug-binding domain of the human multidrug resistance P-gp likely consists of residues from multiple transmembrane (TM) segments. In this study, we completed cysteine-scanning mutagenesis of all the predicted TM segments of P-gp (TMs 1-5 and 7-10) and tested for inhibition by a thiol-reactive substrate (dibromobimane) to identify residues within the drug binding domain. The activities of 189 mutants were analyzed. Verapamil-stimulated ATPase activities of seven mutants (Y118C and V125C (TM2), S222C (TM4), I306C (TM5), S766C (TM9), and I868C and G872C (TM10)) were inhibited by more than 50% by dibromobimane. The activities of mutants S222C (TM4), I306C (TM5), I868C (TM10), and G872C (TM10), but not that of mutants Y118C (TM2), V125C (TM2), and S776C (TM9), were protected from inhibition by dibromobimane by pretreatment with verapamil, vinblastine, or colchicine. These results and those from previous studies (Loo, T. W. and Clarke, D. M. (1997) J. Biol. Chem. 272, 31945-31948; Loo, T. W. and Clarke, D. M. (1999) J. Biol. Chem. 274, 35388-35392) indicate that the drug-binding domain of P-gp consists of residues in TMs 4, 5, 6, 10, 11, and 12. PMID- 11013261 TI - The immunophilin-like protein XAP2 regulates ubiquitination and subcellular localization of the dioxin receptor. AB - The dioxin (aryl hydrocarbon) receptor is a ligand-dependent transcription factor that induces expression of a number of genes encoding drug metabolizing enzymes. The nonactivated form of the dioxin receptor is associated with heat shock protein (hsp) 90, the co-chaperone p23, and the immunophilin-like protein XAP2. Whereas hsp90 has a role in maintenance of the high-affinity ligand binding conformation of the dioxin receptor complex, and p23 stabilizes receptor-hsp90 interaction, the exact role of XAP2 is largely unknown. Here we show that XAP2 protected the ligand-free form of receptor against ubiquitination, resulting in increased dioxin receptor protein levels. Upon exposure to ligand, nuclear translocation of the dioxin receptor was markedly delayed by XAP2, indicating an additional role of XAP2 in regulation of the subcellular localization of the receptor by a mechanism of cytoplasmic retention. In order to mediate these effects, XAP2 required stable association with the hsp90-p23 molecular chaperone complex. The association of XAP2 as well as p23 with the dioxin receptor was determined by the functional state of hsp90. These data indicate a novel mode of regulation of dioxin receptor signaling by the hsp90-dependent molecular chaperone machinery. PMID- 11013262 TI - Protein-tyrosine phosphatase D1, a potential regulator and effector for Tec family kinases. AB - Etk, also named Bmx, is a member of the Tec tyrosine kinase family, which is characterized by a multimodular structure including a pleckstrin homology (PH) domain, an SH3 domain, an SH2 domain, and a catalytic domain. The signaling mechanisms regulating Etk kinase activity remain largely unknown. To identify factor(s) regulating Etk activity, we used the PH domain and a linker region of Etk as a bait for a yeast two-hybrid screen. Three independent clones encoding protein-tyrosine phosphatase D1 (PTPD1) fragments were isolated. The binding of PTPD1 to Etk is specific since PTPD1 cannot associate with either the Akt PH domain or lamin. In vitro and in vivo binding studies demonstrated that PTPD1 can interact with Etk and that residues 726-848 of PTPD1 are essential for this interaction. Deletion analysis of Etk indicated that the PH domain is essential for PTPD1 interaction. Furthermore, the Etk-PTPD1 interaction stimulated the kinase activity of Etk, resulting in an increased phosphotyrosine content in both factors. The Etk-PTPD1 interaction also increased Stat3 activation. The effect of PTPD1 on Etk activation is specific since PTPD1 cannot potentiate Jak2 activity upon Stat3 activation. In addition, Tec (but not Btk) kinase can also be activated by PTPD1. Taken together, these findings indicate that PTPD1 can selectively associate with and stimulate Tec family kinases and modulate Stat3 activation. PMID- 11013263 TI - A novel nuclear receptor corepressor complex, N-CoR, contains components of the mammalian SWI/SNF complex and the corepressor KAP-1. AB - Transcriptional silencing by many transcription factors is mediated by the nuclear receptor corepressor (N-CoR). The mechanism by which N-CoR represses basal transcription involves the direct or indirect recruitment of histone deacetylases (HDACs). We have isolated two multiprotein N-CoR complexes, designated N-CoR-1 and N-CoR-2, which possess histone deacetylase activity that is mediated by distinct HDACs. Based on Western blotting using antibodies against known subunits, the only HDAC found in the N-CoR-1 complex was HDAC3. In contrast, N-CoR-2 contained predominantly HDAC1 and HDAC2 as well as several other subunits that are found in the Sin3A.HDAC complex. Using mass spectrometry and Western blotting, we have identified several novel components of the N-CoR-1 complex including the SWI/SNF-related proteins BRG1, BAF 170, BAF 155, BAF 47/INI1, and the corepressor KAP-1 that is involved in silencing heterochromatin. Indirect immunofluorescence has revealed that both KAP-1 and N-CoR colocalize throughout the nucleus. These results suggest that N-CoR is found in distinct multiprotein complexes, which are involved in multiple pathways of transcriptional repression. PMID- 11013264 TI - Use of diethyl(2-methylpyrrolidin-2-yl)phosphonate as a highly sensitive extra- and intracellular 31P NMR pH indicator in isolated organs. Direct NMR evidence of acidic compartments in the ischemic and reperfused rat liver. AB - The novel phosphorylated pyrrolidine diethyl(2-methylpyrrolidin-2-yl)phosphonate (DEPMPH) was evaluated as a (31)P NMR probe of the pH changes associated with ischemia/reperfusion of rat isolated hearts and livers. In vitro titration curves indicated that DEPMPH exhibited a 4-fold larger amplitude of chemical shift variation than inorganic phosphate yielding an enhanced NMR sensitivity in the pH range of 5.0-7.5 that allowed us to assess pH variations of less than 0.1 pH units. At the non-toxic concentration of 5 mm, DEPMPH distributed into external and cytosolic compartments in both normoxic organs, as assessed by the appearance of two resonance peaks. An additional peak was observed in normoxic and ischemic livers, assigned to DEPMPH in acidic vesicles (pH 5.3-5.6). During severe myocardial ischemia, a third peak corresponding to DEPMPH located in ventricular and atrial cavities appeared (pH 6.9). Mass spectrometry and NMR analyses of perchloric extracts showed that no significant metabolism of DEPMPH occurred in the ischemic liver. Reperfusion with plain buffer resulted in a rapid washout of DEPMPH from both organs. It was concluded that the highly pH-sensitive DEPMPH could be of great interest in noninvasive ex vivo studies of pH gradients that may be involved in many pathological processes. PMID- 11013265 TI - Fate of endogenously synthesized cholesterol in Niemann-Pick type C1 cells. AB - Mammalian cells obtain cholesterol via two pathways: endogenous synthesis in the endoplasmic reticulum and exogenous sources mainly through the low density lipoprotein (LDL) receptor pathway. We performed pulse-chase experiments to monitor the fate of endogenously synthesized cholesterol and showed that, after reaching the plasma membrane from the endoplasmic reticulum, the newly synthesized cholesterol eventually accumulates in an internal compartment in Niemann-Pick type C1 (NPC1) cells. Thus, the ultimate fate of endogenously synthesized cholesterol in NPC1 cells is the same as LDL-derived cholesterol. However, the time required for endogenous cholesterol to accumulate internally is much slower than LDL-derived cholesterol. Different pathways thus govern the post plasma membrane trafficking of endogenous cholesterol and LDL-derived cholesterol to the internal compartment. Results using the inhibitor N-butyldeoxynojirimycin, which depletes cellular complex glycosphingolipids, demonstrates that the cholesterol trafficking defect in NPC1 cells is not caused by ganglioside accumulation. The ultimate cause of death in NPC disease is progressive neurological deterioration in the central nervous system, where the major source of cholesterol is derived from endogenous synthesis. Our current study provides a plausible link between defects in intracellular trafficking of endogenous cholesterol and the etiology of Niemann-Pick type C disease. PMID- 11013266 TI - Inhibition of Trypanosoma brucei gene expression by RNA interference using an integratable vector with opposing T7 promoters. AB - RNA interference is a powerful method for inhibition of gene expression in Trypanosoma brucei (Ngo, H., Tschudi, C., Gull, K., and Ullu, E. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 14687-14692). Here we describe a vector (pZJM) for in vivo tetracycline-inducible synthesis of double-stranded RNA (dsRNA) in stably transformed cells. The dsRNA is synthesized from opposing T7 promoters. We tested the vector with genes involved in processes such as kinetoplast DNA replication, mitochondrial mRNA synthesis, glycosyl phosphatidylinositol biosynthesis, glycosome biogenesis, and polyamine biosynthesis. In most cases the induction of dsRNA caused specific and dramatic loss of the appropriate mRNA, and in many cases there was growth inhibition or cell death. One striking phenotype was the loss of kinetoplast DNA after interference with expression of a topoisomerase II. The gene being analyzed by this procedure need not even be fully sequenced. In fact, many of the genes we tested were derived from partial sequences in the T. brucei genome data base that were identified by homology with known proteins. It takes as little as 3 weeks from identification of a gene sequence in the data base to the appearance of a phenotype. PMID- 11013268 TI - Perlecan heparan sulfate proteoglycan. A novel receptor that mediates a distinct pathway for ligand catabolism PMID- 11013267 TI - Nalpha -terminal acetylation of eukaryotic proteins. PMID- 11013269 TI - Identification and cloning of two histone fold motif-containing subunits of HeLa DNA polymerase epsilon PMID- 11013270 TI - Epstein-Barr virus, the CNS, and AIDS-related lymphomas: as close as flame to smoke. PMID- 11013271 TI - Epstein-Barr virus infection is predictive of CNS involvement in systemic AIDS related non-Hodgkin's lymphomas. AB - PURPOSE: This study aimed at correlating Epstein-Barr virus (EBV) infection of systemic AIDS-related non-Hodgkin lymphomas (AIDS-NHL) with the development of a CNS localization of the tumor. PATIENTS AND METHODS: Demographic, epidemiologic, clinical, histologic, and virologic features were collected for all systemic AIDS NHL patients included in the study (n = 50). Pathologic specimens were classified according to the working formulation for NHL and the Revised European-American Lymphoma classification. EBV infection in tumor tissue samples was studied by EBV small encoded RNA in situ hybridization; EBV-DNA detection in CSF was carried out by nested polymerase chain reaction using Epstein-Barr nuclear antigen-1-specific primers. In addition, selected EBV-positive lymphomas were subjected to a detailed characterization of EBV molecular heterogeneity. RESULTS: Eleven patients had a CNS involvement at some point during their clinical history (four at diagnosis and seven at relapse). Thirty patients (11 with CNS involvement and 19 without) harbored EBV infection of the tumor. Sensitivity, specificity, and positive and negative predictive values of EBV-DNA detection in CSF for CNS involvement by lymphoma were 90%, 100%, 100%, and 97.6%, respectively. Factors significantly predictive of CNS involvement were EBV infection of the tumor (P=.003), an extranodal disease at diagnosis other than CNS (P=.006), and a non CNS relapse (P=.01). In four cases of CNS involvement, EBV-DNA in CSF preceded any other sign of disease by a mean of 35 days. CONCLUSION: These results show that EBV infection of the tumor clone significantly increases the risk of CNS involvement by systemic AIDS-NHL, without regard of specific molecular features. The detection of EBV-DNA in the CSF of AIDS-NHL patients may select cases with higher risk of CNS involvement and, therefore, may prove useful in the therapeutic stratification of these tumors. PMID- 11013272 TI - Expression of interferon regulatory factor 4 in chronic myeloid leukemia: correlation with response to interferon alfa therapy. AB - PURPOSE: Mice experiments have established an important role for interferon regulatory factor (IRF) family members in hematopoiesis. We wanted to study the expression of interferon regulatory factor 4 (IRF4) in various hematologic disorders, especially chronic myeloid leukemia (CML), and its association with response to interferon alfa (IFN-alpha) treatment in CML. MATERIALS AND METHODS: Blood samples from various hematopoietic cell lines, different leukemia patients (70 CML, 29 acute myeloid leukemia [AML], 10 chronic myelomonocytic leukemia [CMMoL], 10 acute lymphoblastic leukemia, and 10 chronic lymphoid leukemia patients), and 33 healthy volunteers were monitored for IRF4 expression by reverse transcriptase polymerase chain reaction. Then, with a focus on CML, the IRF4 level was determined in sorted cell subpopulations from CML patients and healthy volunteers and in in vitro-stimulated CML cells. Furthermore, IRF4 expression was compared in the CML samples taken before IFN-alpha therapy and in 47 additional CML samples taken during IFN-alpha therapy. IRF4 expression was then correlated with cytogenetic response to IFN-alpha. RESULTS: IRF4 expression was significantly impaired in CML, AML, and CMMoL samples. The downregulation of IRF4 in CML samples was predominantly found in T cells. In CML patients during IFN-alpha therapy, a significant increase in IRF4 levels was detected, and this was also observed in sorted T cells from CML patients. The increase seen during IFN-alpha therapy was not due to different blood counts. In regard to the cytogenetic response with IFN-alpha, a good response was associated with high IRF4 expression. CONCLUSION: IRF4 expression is downregulated in T cells of CML patients, and its increase is associated with a good response to IFN-alpha therapy. These data suggest IRF4 expression as a useful marker to monitor, if not predict, response to IFN-alpha in CML. PMID- 11013273 TI - Phase III randomized trial of amifostine as a radioprotector in head and neck cancer. AB - PURPOSE: Radiotherapy for head and neck cancer causes acute and chronic xerostomia and acute mucositis. Amifositine and its active metabolite, WR-1065, accumulate with high concentrations in the salivary glands. This randomized trial evaluated whether amifostine could ameliorate these side effects without compromising the effectiveness of radiotherapy in these patients. PATIENTS AND METHODS: Patients with previously untreated head and neck squamous cell carcinoma were eligible. Primary end points included the incidence of grade > or =2 acute xerostomia, grade > or =3 acute mucositis, and grade > or =2 late xerostomia and were based on the worst toxicity reported. Amifostine was administered (200 mg/m(2) intravenous) daily 15 to 30 minutes before irradiation. Radiotherapy was given once daily (1.8 to 2.0 Gy) to doses of 50 to 70 Gy. Whole saliva production was quantitated preradiotherapy and regularly during follow-up. Patients evaluated their symptoms through a questionnaire during and after treatment. Local-regional control was the primary antitumor efficacy end point. RESULTS: Nausea, vomiting, hypotension, and allergic reactions were the most common side effects. Fifty-three percent of the patients receiving amifostine had at least one episode of nausea and/or vomiting, but it only occurred with 233 (5%) of 4,314 doses. Amifostine reduced grade > or =2 acute xerostomia from 78% to 51% (P<.0001) and chronic xerostomia grade > or = 2 from 57% to 34% (P=.002). Median saliva production was greater with amifostine (0.26 g v 0.10 g, P=.04). Amifostine did not reduce mucositis. With and without amifostine, 2-year local regional control, disease-free survival, and overall survival were 58% versus 63%, 53% versus 57%, and 71% versus 66%, respectively. CONCLUSION: Amifostine reduced acute and chronic xerostomia. Antitumor treatment efficacy was preserved. PMID- 11013275 TI - Pretreatment nomogram for predicting the outcome of three-dimensional conformal radiotherapy in prostate cancer. AB - PURPOSE: Several studies have defined risk groups for predicting the outcome after external-beam radiotherapy of localized prostate cancer. However, most models formed patient risk groups, and none of these models considers radiation dose as a predictor variable. The purpose of this study was to develop a nomogram to improve the accuracy of predicting outcome after three-dimensional conformal radiotherapy. MATERIALS AND METHODS: This study was a retrospective, nonrandomized analysis of patients treated at the Memorial Sloan-Kettering Cancer Center between 1988 and 1998. Clinical parameters of the 1,042 patients included stage, biopsy Gleason score, pretreatment serum prostate-specific antigen (PSA) level, whether neoadjuvant androgen deprivation therapy was administered, and the radiation dose delivered. Biochemical (PSA) treatment failure was scored when three consecutive rises of serum PSA occurred. A nomogram, which predicts the probability of remaining free from biochemical recurrence for 5 years, was validated internally on this data set using a bootstrapping method and externally using a cohort of patients treated at the Cleveland Clinic, Cleveland, OH. RESULTS: When predicting outcomes for patients in the validation data set from the Cleveland Clinic, the nomogram had a Somers' D rank correlation between predicted and observed failure times of 0.52. Predictions from this nomogram were more accurate (P<.0001) than the best of seven published risk stratification systems, which achieved a Somers' D coefficient of 0.47. CONCLUSION: The development process illustrated here produced a nomogram that seems to predict more accurately than other available systems and may be useful for treatment selection by both physicians and patients. PMID- 11013274 TI - High-dose chemotherapy as initial salvage chemotherapy in patients with relapsed testicular cancer. AB - PURPOSE: To assess the role of high-dose chemotherapy as initial salvage chemotherapy in patients with relapsed testicular cancer. PATIENTS AND METHODS: From August 1992 to April 1998, 65 patients with testicular cancer were treated with high-dose carboplatin and etoposide followed by peripheral-blood stem-cell transplantation or autologous bone marrow transplantation rescue as initial salvage chemotherapy at Indiana University. An identical course was given after hematopoietic reconstitution. Postchemotherapy resection of residual disease was performed in selected patients with incomplete radiographic response associated with normalization of markers. The median follow-up was 39 months (range, 16 to 91 months). RESULTS: Thirty-seven (57%) of the 65 patients are continuously disease-free. Three additional patients are disease-free with subsequent surgery. High-dose chemotherapy was associated with significant morbidity but no treatment related mortality. CONCLUSION: High-dose chemotherapy as initial salvage chemotherapy achieved impressive long-term survival with acceptable toxicity in patients with relapsed testicular cancer. PMID- 11013276 TI - Effect of radiotherapy after breast-conserving treatment in women with breast cancer and germline BRCA1/2 mutations. AB - PURPOSE: Recent laboratory data suggest a role for BRCA1/2 in the cellular response to DNA damage. There is a paucity of clinical data, however, examining the effect of radiotherapy (RT), which causes double-strand breaks, on breast tissue from BRCA1/2 mutation carriers. Thus the goals of this study were to compare rates of radiation-associated complications, in-breast tumor recurrence, and distant relapse in women with BRCA1/2 mutations treated with breast conserving therapy (BCT) using RT with rates observed in sporadic disease. PATIENTS AND METHODS: Seventy-one women with a BRCA1/2 mutation and stage I or II breast cancer treated with BCT were matched 1:3 with 213 women with sporadic breast cancer. Conditional logistic regression models were used to compare matched cohorts for rates of complications and recurrence. RESULTS: Tumors from women in the genetic cohort were associated with high histologic (P =.0004) and nuclear (P =.009) grade and negative estrogen (P=.0001) and progesterone (P=.002) receptors compared with tumors from the sporadic cohort. Using Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer toxicity scoring, there were no significant differences in acute or chronic morbidity in skin, subcutaneous tissue, lung, or bone. The 5-year actuarial overall survival, relapse-free survival, and rates of tumor control in the treated breast for the patients in the genetic cohort were 86%, 78%, and 98%, respectively, compared with 91%, 80%, and 96%, respectively, for the sporadic cohort (P = not significant). CONCLUSION: There was no evidence of increased radiation sensitivity or sequelae in breast tissue heterozygous for a BRCA1/2 germline mutation compared with controls, and rates of tumor control in the breast and survival were comparable between BRCA1/2 carriers and controls at 5 years. Although additional follow-up is needed, these data may help in discussing treatment options in the management of early-stage hereditary breast cancer and should provide reassurance regarding the safety of administering RT to carriers of a germline BRCA1/2 mutation. PMID- 11013277 TI - Phase II study of vinorelbine with protracted fluorouracil infusion as a second- or third-line approach for advanced breast cancer patients previously treated with anthracyclines. AB - PURPOSE: To evaluate the feasibility and activity of vinorelbine in association with protracted infusional fluorouracil in patients with advanced breast cancer who were previously treated with anthracycline-containing regimens. PATIENTS AND METHODS: Eighty-three consecutive patients were entered onto the study. Forty three patients experienced treatment failure or relapse after anthracycline based, first-line chemotherapy for advanced disease and 29 experienced treatment failure or relapse after first- and second-line approaches; 11 patients experienced progressive disease within 6 months of completion of adjuvant anthracycline therapy. Sites of involvement were as follows: liver involvement, 42 patients (50.6%); lung 24 (28.9%); bone, 49 (59.0%); and skin/lymph nodes, 21 (25.3%). Treatment consisted of vinorelbine 30 mg/m(2) administered on days 1 and 15 every 28 days and fluorouracil 200 mg/m(2)/d given continuously over a 24-hour period. RESULTS: Toxicity was recorded for 441 cycles. The scheme was well tolerated: grade 1/2 nausea/vomiting occurred in 13 patients (15.6%), grade 1/2 diarrhea in nine (10.8%), and grade 2/3 stomatitis in six (7.2%). Three patients (3.6%) experienced grade 3/4 leukopenia and four (4.8%) experienced grade 2/3 anemia. Grade 2/3 neurologic toxicity was observed in three cases (3.6%), and grade 2/3 hand-foot syndrome was observed in three (3.6%). The median relative dose-intensity was 92% and 100% for vinorelbine and fluorouracil, respectively. Six patients (7.2%) attained a complete clinical response and 45 (54.2%) attained a partial response, for an overall response rate of 61.4% (95% confidence interval, 50.9% to 71.9%). Twenty-one patients (25.3%) obtained disease stabilization, and 11 (13.3%) experienced disease progression. Median time to progression in responding patients was 15 months; median overall survival of the entire population was 22 months. CONCLUSION: Vinorelbine associated with protracted infusional fluorouracil is an active and manageable scheme in advanced breast cancer patients previously treated with anthracyclines. The response obtained is durable. PMID- 11013278 TI - Impact of neoadjuvant chemotherapy on postoperative morbidity in soft tissue sarcomas. AB - PURPOSE: The purpose of this study was to test the hypothesis that neoadjuvant chemotherapy (NeoCT) does not increase morbidity in patients undergoing radical surgery for soft tissue sarcomas. PATIENTS AND METHODS: The records of 309 patients who presented to The University of Texas M.D. Anderson Cancer Center for definitive surgical management of primary soft tissue sarcomas were retrospectively reviewed. One hundred five patients who received NeoCT were compared with 204 patients who had surgery first (Surg). Patients had extremity sarcomas (71 NeoCT patients and 130 Surg patients) or retroperitoneal/visceral sarcomas (34 NeoCT and 74 Surg). RESULTS: NeoCT patients had larger tumors (median, 12 v 8 cm), more frequently had high-grade tumors (90% v 64%), and were younger (median age 47 v 55 years). The incidence of surgical complications was not different for NeoCT patients than for Surg patients, both in those with extremity sarcomas (34% v 41%) and in those with retroperitoneal/visceral sarcomas (29% v 34%). The most common complications were wound infections and other wound complications. Preoperative radiation therapy, autologous flap coverage, and extremity tumors were associated with increased wound complications. No significant differences in length of hospital stay, rate of readmission, or rate of reoperation for complications were found between the NeoCT and Surg groups. One of the three postoperative deaths in our series occurred in the NeoCT group. CONCLUSION: In this retrospective review, there was no evidence that NeoCT increased postoperative morbidity in patients with soft tissue sarcomas. Prospective, randomized studies are needed to confirm these results. PMID- 11013279 TI - Eastern Cooperative Oncology Group Phase I trial of protracted venous infusion fluorouracil plus weekly gemcitabine with concurrent radiation therapy in patients with locally advanced pancreas cancer: a regimen with unexpected early toxicity. AB - PURPOSE: We performed a phase I trial of protracted venous infusion (PVI) fluorouracil (5-FU) plus weekly gemcitabine with concurrent radiation therapy in patients with locally advanced pancreas cancer to determine the maximum-tolerated dose of gemcitabine that could be safely administered. We also sought to identify the toxicities associated with this treatment protocol. PATIENTS AND METHODS: Seven patients with locally advanced pancreas cancer were treated with planned doses of radiation (59.4 Gy) and PVI of 5-FU (200 mg/m(2)/d) with gemcitabine doses of 50 to 100 mg/m(2)/wk. RESULTS: Two of three patients at the 100 mg/m(2)/wk dose level experienced dose-limiting toxicity (DLT), as did three of four at the 50-mg/m(2)/wk dose level. One patient experienced a mucocutaneous reaction described as a Stevens-Johnson syndrome that was attributed to chemotherapy. Three patients developed gastric or duodenal ulcers with severe bleeding requiring transfusion. One patient developed severe thrombocytopenia lasting longer than 4 weeks. Three of the five episodes of DLT developed at radiation doses < or = 36 Gy. CONCLUSION: Based on this experience, we cannot recommend further investigation of regimens incorporating gemcitabine into regimens of radiation with PVI 5-FU. The mechanism of this synergistic toxicity remains to be determined. PMID- 11013280 TI - Phase III comparative study of high-dose cisplatin versus a combination of paclitaxel and cisplatin in patients with advanced non-small-cell lung cancer. AB - PURPOSE: New effective chemotherapy is needed to improve the outcome of patients with advanced non-small-cell lung cancer (NSCLC). Paclitaxel administered as a single agent or in combination with cisplatin has been shown to be a potentially new useful agent for the treatment of NSCLC. PATIENTS AND METHODS: Between January 1995 and April 1996, 414 patients with stage IIIB or IV NSCLC were randomized to received either a control arm of high-dose cisplatin (100 mg/m(2)) or a combination of paclitaxel (175 mg/m(2), 3-hour infusion) and cisplatin (80 mg/m(2)) every 21 days. RESULTS: Compared with the cisplatin-only arm, there was a 9% improvement (95% confidence interval, 0% to 19%) in overall response rate for the paclitaxel/cisplatin arm (17% v 26%, respectively; P=.028). Median time to progression was 2.7 and 4.1 months in the control and paclitaxel/cisplatin arm, respectively (P=.026). The study, however, failed to show a significant improvement in median survival for the paclitaxel/cisplatin arm (8.6 months in the control arm v 8.1 months in the paclitaxel/cisplatin arm, P=.862). There was more hematotoxicity, peripheral neuropathy, and arthralgia/myalgia on the paclitaxel/cisplatin arm, whereas the high-dose cisplatin arm produced more ototoxicity, nausea, vomiting, and nephrotoxicity. Quality of life (QOL) was similar overall between the two arms. CONCLUSION: This large randomized phase III trial failed to show a significant improvement in survival for the paclitaxel/cisplatin combination compared with high-dose cisplatin in patients with advanced NSCLC. However, the paclitaxel/cisplatin combination did produce a better clinical response, resulting in an increased time to progression while providing a similar QOL. PMID- 11013281 TI - Treatment of brain metastases of small-cell lung cancer: comparing teniposide and teniposide with whole-brain radiotherapy--a phase III study of the European Organization for the Research and Treatment of Cancer Lung Cancer Cooperative Group. AB - PURPOSE: Approximately 60% of patients with small-cell lung cancer (SCLC) develop brain metastases. Whole-brain radiotherapy (WBRT) gives symptomatic improvement in more than 50% of these patients. Because brain metastases are a sign of systemic progression, and chemotherapy was found to be effective as well, it becomes questionable whether WBRT is the only appropriate therapy in this situation. PATIENTS AND METHODS: In a phase III study, SCLC patients with brain metastases were randomized to receive teniposide with or without WBRT. Teniposide 120 mg/m(2) was given intravenously three times a week, every 3 weeks. WBRT (10 fractions of 3 Gy) had to start within 3 weeks from the start of chemotherapy. Response was measured clinically and by computed tomography of the brain. RESULTS: One hundred twenty eligible patients were randomized. A 57% response rate was seen in the combined-modality arm (95% confidence interval [CI], 43% to 69%), and a 22% response rate was seen in the teniposide-alone arm (95% CI, 12% to 34%) (P<.001). Time to progression in the brain was longer in the combined modality group (P=.005). Clinical response and response outside the brain were not different. The median survival time was 3.5 months in the combined-modality arm and 3.2 months in the teniposide-alone arm. Overall survival in both groups was not different (P=.087). CONCLUSION: Adding WBRT to teniposide results in a much higher response rate of brain metastases and in a longer time to progression of brain metastases than teniposide alone. Survival was poor in both groups and not significantly different. PMID- 11013282 TI - Contribution of dexamethasone to control of chemotherapy-induced nausea and vomiting: a meta-analysis of randomized evidence. AB - PURPOSE: To synthesize the available randomized evidence on the efficacy of dexamethasone when used for protection against acute and delayed nausea and vomiting in patients receiving highly or moderately emetogenic cancer chemotherapy. MATERIALS AND METHODS: A meta-analysis was performed using trials identified through MEDLINE (1966 to April 1999), Embase, Derwent Drug File, and the Cochrane Library's Database of Controlled Trials. Data on acute and delayed emesis and nausea were collected. All randomized studies comparing dexamethasone to placebo, no treatment, or other antiemetics qualified, including cross-over trials providing first-cycle data. RESULTS: Of 1,200 citations screened, 32 studies with 42 pertinent comparisons and 5,613 patients were included in the meta-analysis. Dexamethasone was superior to placebo or no treatment for complete protection from acute emesis (odds ratio, 2.22; 95% confidence interval [CI], 1.89 to 2.60) and for complete protection from delayed emesis (odds ratio, 2.04; 95% CI, 1.63 to 2.56). The results were similar for complete protection from nausea. The pooled risk difference for complete protection from emesis was 16% for both the acute and delayed phases (95% CI, 13% to 19% and 11% to 20%, respectively). The beneficial effect was similar in subgroups defined by various study design parameters. No trial addressed the efficacy of dexamethasone in the delayed phase without having administered dexamethasone for acute-phase protection as well. CONCLUSION: Dexamethasone is clearly effective in protecting from emesis both in the acute and delayed phases, with emesis avoided in one patient out of six treated. Future trials should determine whether the delayed phase effect is independent of the acute-phase benefit. PMID- 11013284 TI - Unusual manifestations of acute leukemia. Case 1. CNS extramedullary relapse of acute promyelocytic leukemia after arsenic trioxide-induced remission. PMID- 11013283 TI - Phase I assessment of the pharmacokinetics, metabolism, and safety of emitefur in patients with refractory solid tumors. AB - PURPOSE: To determine the toxicities, dose-limiting toxicities (DLT), maximum tolerated dose, and pharmacokinetic profile of emitefur (BOF-A2) in patients with advanced solid tumors. METHODS: This was a phase I dose-escalating trial in which cohorts of patients received BOF-A2 (cohort 1, 300 mg/m(2) orally [PO] tid; cohort 2, 200 mg/m(2) PO tid; cohort 3, 200 mg/m(2) bid; and cohort 4, 250 mg/m(2) bid) for 14 consecutive days followed by 1 week of rest (cycle 1). Pharmacokinetics, toxicity, and tumor response were monitored. RESULTS: Nineteen patients received 110 cycles (three patients in cohort 1, three patients in cohort 2, 10 patients in cohort 3, and three patients in cohort 4). DLT (grade 3 stomatitis, diarrhea, leukopenia) was observed in cohorts 1, 2, and 4. Pharmacokinetics indicated that prolonged systemic expression of fluorouracil (5 FU) is maintained after administration of BOF-A2 at a dose of 200 mg bid for 14 days. The mean steady-state concentration of plasma 5-FU was > or = 24 ng/mL, which was 184-fold greater than the minimum effective cytotoxic concentration in vitro. Lack of variation of 5-FU trough levels within a day at steady-state indicates suppression of circadian variation. One patient in cohort 3 achieved a partial response and five patients maintained stable disease in excess of 6 months. CONCLUSION: BOF-A2 at a dose of 200 mg PO bid for 14 days followed by 7 days of rest is well tolerated. Prolonged exposure to 5-FU above the predicted preclinical minimum effective concentration is maintained, without evidence of circadian variation. Furthermore, evidence of antitumor activity is suggested. PMID- 11013285 TI - Unusual manifestations of acute leukemia. Case 2. Leukemia and rash: paraneoplastic or drug-induced? PMID- 11013286 TI - Unusual manifestations of acute leukemia. Case 3. Unilateral leukemic hypopyon. PMID- 11013287 TI - The art of oncology: when the tumor is not the target. Tell it like it is. PMID- 11013288 TI - Myelodysplasia after autotransplantation. PMID- 11013290 TI - Radiation therapy or surgery for base-of-tongue tumors? PMID- 11013289 TI - Problematic WHO reclassification of myelodysplastic syndromes. Members of the International MDS Study Group. PMID- 11013291 TI - Paclitaxel for limited-stage small-cell lung cancer. PMID- 11013292 TI - Thalidomide in the treatment of high-grade gliomas. PMID- 11013293 TI - Prophylactic surgery: oophorectomy or adnexectomy? PMID- 11013294 TI - Stage shift and complementary/alternative medicine. PMID- 11013295 TI - Urinary alpha-tocopherol metabolites in alpha-tocopherol transfer protein deficient patients. AB - Patients with alpha-tocopherol transfer protein (alpha-TTP) defects experience neurological symptoms characteristic of vitamin E deficiency and depend on continuous high alpha-tocopherol supplements. We investigated the excretion of 2,5,7, 8-tetramethyl-2(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC), a urinary metabolite of alpha-tocopherol, as a putative marker for the alpha-tocopherol status of alpha-TTP-deficient patients and control subjects. In three patients vitamin E supplementation was stopped for short periods of time, during which plasma alpha-tocopherol concentrations and urinary alpha-CEHC excretion were measured. In the patients, plasma alpha-tocopherol decreased below normal (<5 micromol/l) but alpha-CEHC excretion remained above the range of unsupplemented control subjects (0.118-0.306 mg/day, n = 6). In healthy subjects, however, alpha CEHC excretion was increased only after surpassing a plasma alpha-tocopherol threshold of 30-40 micromol/l. Such a threshold did not exist in patients. The general mechanism of alpha-tocopherol degradation did not appear to differ between patients and control subjects. The presumed mechanism of omega- and subsequent beta-oxidation was supported by the detection of alpha- CPHC, an alpha -CEHC homolog with a side chain longer by 3 carbon atoms, both in supplemented patients and in control subjects. PMID- 11013297 TI - Structural and biosynthetic studies of a principal bile alcohol, 27-nor-5beta cholestane-3alpha,7alpha,12alpha,24,25-pentol, in human urine. AB - The stereochemistry at C-24 and C-25 of 27-nor-5beta-cholestane 3alpha,7alpha,12alpha,24 ,25-pentol, a principal bile alcohol in human urine, and its biosynthesis are studied. Four stereoisomers of the C(26)-24,25-pentols were synthesized by reduction with LiAlH(4) of the corresponding epoxides prepared from (24S)- or (24R)-27-nor-5beta-cholest-25-ene-3alpha, 7alpha,12alpha,24 tetrol. The stereochemistries at C-25 were deduced by comparison of the C(26) 24,25-pentols with the oxidation products of (24Z)-27-nor-5beta-cholest-24-ene 3alpha,7alpha, 12alpha-triol with osmium tetraoxide. On the basis of this assignment, the principal bile alcohol excreted into human and rat urine was determined to be (24S,25R)-27-nor-5beta-cholestane-3alpha,7alpha, 12alpha,24,25 pentol, accompanied by a lesser amount of (24R, 25R)-isomer. To elucidate the biosynthesis of the C(26)-24,25-pentol, a putative intermediate, 3alpha,7alpha, 12alpha-trihydroxy-27-nor-5beta-cholestan-24-one, derived from 3alpha,7alpha, 12alpha-trihydroxy-24-oxo-5beta-cholestanoic acid by decarboxylation during the side-chain oxidation of 3alpha,7alpha, 12alpha-trihydroxy-5beta-cholestanoic acid, was incubated with rat liver homogenates. The 24-oxo-bile alcohol could be efficiently reduced to yield mainly (24R)-27-nor-5beta-cholestane-3alpha,7alpha, 12alpha,24-tetrol. If a 25R-hydroxylation of the latter steroid occurs, it should lead to formation of (24S,25R)-C(26)-24,25-pentol. Now it has appeared that a major bile alcohol excreted into human urine is (24S,25R)-27-nor-5beta-cholestane 3alpha,7alpha, 12alpha, 24, 25-pentol, which might be derived from 3alpha,7alpha, 12alpha-trihydroxy-27-nor-5beta-cholestan-24-one via (24R)-27-nor-5beta cholestane-3alpha, 7alpha,12alpha,24-tetrol. PMID- 11013296 TI - Age-related decrease of dehydroepiandrosterone concentrations in low density lipoproteins and its role in the susceptibility of low density lipoproteins to lipid peroxidation. AB - The incidence of atherosclerosis and related diseases increases with age. The aging process may enhance lipoprotein modification, which leads to an increase in the susceptibility of low density lipoprotein (LDL) and high density lipoprotein (HDL) to oxidation. Dehydroepiandrosterone (DHEA), the most abundant steroid hormone in humans, has been shown to have antiatherogenic effects. This hormone also decreases dramatically with age. In the present study, we were interested in determining the presence of DHEA/DHEAS (dehydroepiandrosterone sulfate) and changes in their concentrations in HDL and LDL lipoproteins with age. Moreover, we studied the susceptibility of LDL to oxidation with age in the presence or absence of vitamin E or DHEA. We demonstrated that vitamin E is unable to restore the decreased resistance to oxidation of LDL from elderly subjects to that of LDL obtained from young subjects. Furthermore, our results provide evidence that DHEA is an integral part of LDL and HDL and disappears to almost nondetectable levels during aging. The DHEA incorporated into the LDL from elderly subjects increased LDL resistance to oxidation in a concentration-dependent manner. The increased resistance provided by DHEA was higher than that with vitamin E. DHEA seems to act either by protecting vitamin E from disappearance from LDL under oxidation or by scavenging directly the free radicals produced during the oxidative process. Our results suggests that DHEA exerts an antioxidative effect on LDL, which could have antiatherogenic consequences. Careful clinical trials of DHEA replacement should determine whether this ex vivo effect could be translated into any measurable antiatherogenic (cardioprotective) action. PMID- 11013298 TI - Polymorphism of class A scavenger receptors in C57BL/6 mice. AB - Scavenger receptors class A (SR-A) have been hypothesized to regulate the development of atherosclerotic lesions through recognition of modified low density lipoprotein (LDL) and macrophage adhesion to substrata. Supporting data have been collected from studies using the monoclonal antibody 2F8, an antibody developed from the BALB/c strain-derived macrophage cell line, RAW.264. Although 2F8 immunostained both cultured peritoneal macrophages (MPM) and thymic macrophages from Swiss, BALB/c, and DBA/2 mice, no immunostaining was detected in cells and tissues from C57BL/6 mice, one of the most commonly used atherosclerosis-susceptible mouse strains. Similarly, 2F8 detected SR-A protein in MPM by Western blotting in all strains except C57BL/6. However, a guinea pig antiserum developed to a fusion protein of the extracellular SR-A domain detected appropriately sized bands in all strains. Incubation with 2F8 antagonized acetylated low-density lipoprotein (AcLDL)-induced cholesterol esterification in MPM from BALB/c, Swiss, and DBA/2 strains but had no effect on MPM from C57BL/6 mice. Sequencing of SR-A cDNA from C57BL/6 mice demonstrated complete identity with published sequence in the collagen-like domain. However, four single-residue substitutions were noted in the alpha-helical coiled-coil domain. Site-directed mutagenesis demonstrated that a single substitution (L168S) in this domain accounted for the loss of 2F8 immunoreactivity. Differing reactivities toward a commonly used monoclonal antibody were used to identify polymorphism of SR-A in C57BL/6 mice. PMID- 11013299 TI - Characterization of the basis of lipoprotein [a] lysine-binding heterogeneity. AB - Although elevated plasma concentrations of lipoprotein [a] (Lp[a]) are considered to be a risk factor for atherosclerosis, the mechanisms by which Lp[a] mediates its pathogenic effects have not been conclusively determined. The apolipoprotein [a] (apo[a]) component of Lp[a] confers unique structural properties to this lipoprotein, including the ability to bind to lysine residues in biological substrates. It has been shown, however, that only a fraction of plasma Lp[a] (Lp[a]-Lys(+)) binds to lysine-Sepharose in vitro. The nature of the non-lysine binding Lp[a] fraction in plasma (Lp[a]-Lys(-)) is currently unknown. In the present study, the Lp[a]-Lys(+) fraction was determined in the plasma of six unrelated individuals; the Lp[a]-Lys(+) fraction in these plasma samples ranged from approximately 37 to approximately 48%. Interestingly, purification of the Lp[a] by density gradient ultracentrifugation followed by gel filtration and ion exchange chromatography resulted in progressive increases in the Lp[a]-Lys(+) fraction. Addition of either purified low density lipoprotein (LDL) or fibronectin to the purified Lp[a] at a 1:1 molar ratio reduced the Lp[a]-Lys(+) fraction (maximal decrease of 34 and 20%, respectively) whereas addition of both fibronectin and LDL to the purified Lp[a] resulted in a further decrease (45% maximally) in this fraction. Similar results were obtained by using a recombinant expression system for apo[a]: addition of a 4-fold molar excess of either LDL or fibronectin to conditioned medium containing metabolically labeled recombinant apo[a] reduced the Lys(+) fraction by 49 and 23%, respectively. Taken together, our data suggest that the lysine-binding heterogeneity of plasma Lp[a] is not primarily an intrinsic property of the lipoprotein, but rather results in large part from its ability to noncovalently associate with abundant plasma components such as LDL and fibronectin. These interactions appear to mask the lysine-binding site in apo[a] kringle IV type 10, which mediates the interaction of Lp[a] with lysine-Sepharose. The contribution of these interactions to the function of Lp[a] in vivo remains to be investigated. PMID- 11013300 TI - Electrospray ionization mass spectrometry analyses of nuclear membrane phospholipid loss after reperfusion of ischemic myocardium. AB - The role of nuclear membrane phospholipids as targets of phospholipases resulting in the generation of nuclear signaling messengers has received attention. In the present study, we have exploited the utility of electrospray ionization mass spectrometry to determine the phospholipid content of nuclei isolated from perfused hearts. Rat heart nuclei contained choline glycerophospholipids composed of palmitoyl and stearoyl residues at the sn-1 position with oleoyl, linoleoyl, and arachidonoyl residues at the sn-2 position. Diacyl molecular species were the predominant molecular subclass in the choline glycerophospholipids, with the balance of the molecular species being plasmalogens. In the ethanolamine glycerophospholipid pool from rat heart nuclei approximately 50% of the molecular species were plasmalogens, which were enriched with arachidonic acid at the sn-2 position. A 50% loss of myocytic nuclear choline and ethanolamine glycerophospholipids was observed in hearts rendered globally ischemic for 15 min followed by 90 min of reperfusion in comparisons with the content of these phospholipids in control perfused hearts. The loss of nuclear choline and ethanolamine glycerophospholipids during reperfusion of ischemic myocardium was partially reversed by the calcium-independent phospholipase A(2) (iPLA(2)) inhibitor bromoenol lactone (BEL), suggesting that the loss of nuclear phospholipids during ischemia/reperfusion is mediated, in part, by iPLA(2). Western blot analyses of isolated nuclei from ischemic hearts demonstrated that iPLA(2) is translocated to the nucleus after myocardial ischemia. Taken toghether, these studies have demonstrated that nuclear phospholipid mass decreases after myocardial ischemia by a mechanism that involves, at least in part, phospholipolysis mediated by iPLA2. PMID- 11013301 TI - Acute effects of low density lipoprotein apheresis on metabolic parameters of apolipoprotein B. AB - Apheresis is a treatment option for patients with severe hypercholesterolemia and coronary artery disease. It is unknown whether such therapy changes kinetic parameters of lipoprotein metabolism, such as apolipoprotein B (apoB) secretion rates, conversion rates, and fractional catabolic rates (FCR). We studied the acute effect of apheresis on metabolic parameters of apoB in five patients with drug-resistant hyperlipoproteinemia, using endogenous labeling with D(3)-leucine, mass spectrometry, and multicompartmental modeling. Patients were studied prior to and immediately after apheresis therapy. The two tracer studies were modeled simultaneously, taking into account the non-steady-state concentrations of apoB. The low density lipoprotein (LDL)-apoB concentration was 120+/-32 mg dl(-1) prior to and 52+/-18 mg dl(-1) immediately after apheresis therapy. The metabolic studies indicate that no change in apoB secretion (13.9+/- 4.9 mg kg(-1) day(-1)) is required to fit the tracer and apoB mass data obtained before and after apheresis and that in four of the five patients the LDL-apoB FCR (0.21+/-0.02 day(-1)) was not altered after apheresis. In one subject the LDL-apoB FCR temporarily increased from 0.22 day(-1) to 0.35 day(-1) after apheresis. The conversion rate of very low density lipoprotein (VLDL)-apoB to LDL-apoB is temporarily decreased from 76 to 51% after apheresis and thus less LDL-apoB is produced after apheresis. We conclude that an acute reduction of LDL-apoB concentration does not affect apoB secretion or LDL-apoB FCR, but that apoB conversion to LDL is temporarily decreased. Thus, in most patients the decreased rate of delivery of neutral lipids or apoB to the liver does not result in an upregulation of LDL receptors or in decreased apoB secretion. PMID- 11013302 TI - Inhibition of cholesteryl ester transfer protein by substituted dithiobisnicotinic acid dimethyl ester: involvement Of a critical cysteine. AB - SC-71952, a substituted analog of dithiobisnicotinic acid dimethyl ester, was identified as a potent inhibitor of cholesteryl ester transfer protein (CETP). When tested in an in vitro assay, the concentration of SC-71952 required for half maximal inhibition was 1 microm. The potency of SC-71952 was enhanced 200-fold by preincubation of the inhibitor with CETP, and was decreased 50-fold by treatment with dithiothreitol. Analogs of SC-71952 that did not contain a disulfide linkage were less potent, did not display time dependency, and were not affected by dithiothreitol treatment. Kinetic and biochemical characterization of the inhibitory process of CETP by SC-71952 suggested that the inhibitor initially binds rapidly and reversibly to a hydrophobic site on CETP. With time, the bound inhibitor irreversibly inactivates CETP, presumably by reacting with one of the free cysteines of CETP. Liquid chromatography/mass spectroscopy (LC/MS) analyses of tryptic digests of untreated or SC-71952-inactivated CETP was used to identify which cysteine(s) were potentially involved in the time-dependent, irreversible component of inactivation by the inhibitor. One disulfide bond, Cys143-Cys184, was unaffected by treatment with the inhibitor. Inactivation of CETP by SC-71952 correlated with a progressive decrease in the abundance of free Cys-13 and Cys 333. Conversion of Cys-13 to alanine had no effect on the rapid reversible component of inactivation by SC-71952. However, it abolished the time-dependent enhancement in potency seen with the inhibitor when using wild-type CETP. These data indicate that Cys-13 is critical for the irreversible inactivation of CETP by SC-71952 and provides support for the structural model that places Cys-13 near the neutral lipid-binding site of CETP. PMID- 11013303 TI - Identification of genes specifically expressed in the accumulated visceral adipose tissue of OLETF rats. AB - The Otsuka Long-Evans Tokushima fatty (OLETF) rat is an animal model of type 2 diabetes, characterized by abdominal obesity, insulin resistance, hypertension, and dyslipidemia. To elucidate the underlying molecular mechanism of obesity and its related complications, we used representational difference analysis and identified the genes more abundantly and specifically expressed in the visceral adipose tissue (VAT) of obese OLETF rats compared with the diabetes-resistant counterpart, that is, Long-Evans Tokushima Otsuka (LETO) rats. By Northern blot analysis, we confirmed the differential expression of 13 genes, including 3 novel genes. The upregulated expression of well-characterized lipid metabolic enzymes, such as lipoprotein lipase, phosphoenolpyruvate carboxykinase, and cholesterol esterase, were observed in VAT of OLETF rats. We demonstrated the differential expression of secreted proteins in VAT of OLETF rats, such as thrombospondin 1 and contrapsin-like protease inhibitor. In contrast to lipid enzymes, the secreted proteins revealed exclusive mRNA expression and they were not detected in VAT of LETO rats. Furthermore, the novel genes OL-16 and OL-64 were also expressed specifically in VAT of OLETF rats and were absent in that of LETO rats and other tissues, including subdermal and brown adipose tissues. The C-terminal partial amino acid sequence of OL-64 revealed that it showed approximately 40% homology with alpha(1)-antitrypsin and it seemed to be a new member of the serine proteinase inhibitor (SERPIN) gene family. VAT of OLEFT rats had a unique gene expression profile, and the accumulated VAT-specific known and novel secreted proteins may play a role(s) in the pathogenesis of obesity and its related complications. PMID- 11013304 TI - Apolipoprotein A4-1/2 polymorphism and response of serum lipids to dietary cholesterol in humans. AB - The response of serum lipids to dietary changes is to some extent an innate characteristic. One candidate genetic factor that may affect the response of serum lipids to a change in cholesterol intake is variation in the apolipoprotein A4 gene, known as the APOA4-1/2 or apoA-IVGln360His polymorphism. However, previous studies showed inconsistent results. We therefore fed 10 men and 23 women with the APOA4-1/1 genotype and 4 men and 13 women with the APOA4-1/2 or 2/2 genotype (carriers of the APOA4-2 allele) two diets high in saturated fat, one containing cholesterol at 12.4 mg/MJ, 136.4 mg/day, and one containing cholesterol at 86.2 mg/MJ, 948.2 mg/day. Each diet was supplied for 29 days in crossover design. The mean response of serum low density lipoprotein cholesterol was 0.44 mmol/l (17 mg/dl) in both subjects with the APOA4-1/1 genotype and in subjects with the APOA4-2 allele [95% confidence interval of difference in response, -0.20 to 0.19 mmol/l (-8 to 7 mg/dl)]. The mean response of high density lipoprotein cholesterol was also similar, 0.10 mmol/l (4 mg/dl), in the two APOA-4 genotype groups [95% confidence interval of difference in response, 0.07 to 0.08 mmol/l (-3 to 3 mg/dl)]. Thus, the APOA4-1/2 polymorphism did not affect the response of serum lipids to a change in the intake of cholesterol in this group of healthy Dutch subjects who consumed a background diet high in saturated fat. Knowledge of the APOA4-1/2 polymorphism is probably not a generally applicable tool for the identification of subjects who respond to a change in cholesterol intake. PMID- 11013306 TI - Influence of phospholipid depletion on the size, structure, and remodeling of reconstituted high density lipoproteins. AB - This study shows that phospholipid depletion has a major impact on the size and structure of spherical, reconstituted high density lipoproteins (rHDL) and their remodeling by cholesteryl ester transfer protein (CETP). Spherical rHDL, 9.2 nm in diameter with a phospholipid/cholesteryl ester/unesterified cholesterol/apolipoprotein A-I (apoA-I) (PL/CE/UC/A-I) molar ratio of 37.3/24.5/4.1/1.0, were depleted progressively of phospholipids by incubation with phospholipase A(2). After 30 min of incubation the PL/CE/UC/A-I molar ratio of the rHDL was 8.0/31.2/4.4/1.0 and their diameter had decreased to 8.0 nm. Comparable changes in rHDL size and composition were also apparent when the incubations were carried out in the presence of other lipoprotein classes and lipoprotein-deficient plasma. The changes in size and composition were not accompanied by the dissociation of apoA-I from the rHDL. Phospholipid depletion did not affect rHDL surface charge or the structure and stability of apoA-I. The remodeling of unmodified and phospholipid-depleted rHDL by CETP was also investigated. When the rHDL were incubated for 3 h with CETP and Intralipid, transfers of core lipids between the phospholipid-depleted rHDL and Intralipid were decreased relative to unmodified rHDL. This difference was no longer apparent when the incubations were extended beyond 3 h. In these incubations apoA I dissociated from the phospholipid-depleted and unmodified rHDL at 3 and 12 h, respectively. At 24 h the respective diameters of the unmodified rHDL and phospholipid-depleted rHDL were 8.0 and 7.8 nm. In conclusion, phospholipid depletion has a major impact on rHDL size and their remodeling by CETP. PMID- 11013305 TI - 24-hydroxycholesterol is a substrate for hepatic cholesterol 7alpha-hydroxylase (CYP7A). AB - (24S)-Hydroxycholesterol is formed from cholesterol in the brain and is important for cholesterol homeostasis in this organ. Elimination of (24S) hydroxycholesterol has been suggested to occur in the liver but little is known about the metabolism of this oxysterol. In the present investigation, we report formation of 7alpha, 24-dihydroxycholesterol in pig and human liver. 7alpha hydroxylase activity toward both isomers of 24-hydroxycholesterol [(24S) and (24R)] was found in a partially purified and reconstituted cholesterol 7alpha hydroxylase (CYP7A) enzyme fraction from pig liver microsomes. In contrast, a purified enzyme fraction of pig liver oxysterol 7alpha-hydroxylase with high activity toward 27-hydroxycholesterol did not show any detectable activity toward 24-hydroxycholesterol. 7alpha-Hydroxylation of 24-hydroxycholesterol was strongly inhibited by 7-oxocholesterol, a known inhibitor of CYP7A. Human CYP7A, recombinantly expressed in Escherichia coli and in simian COS cells, showed 7alpha-hydroxylase activity toward both cholesterol and the two isomers of 24 hydroxycholesterol, with a preference for the (24S)-isomer. Our results show that 24-hydroxycholesterol is metabolized by CYP7A, an enzyme previously considered to be specific for cholesterol and cholestanol and not active toward oxysterols. Because CYP7A is the rate-limiting enzyme in the major pathway of bile acid biosynthesis, the possibility is discussed that at least part of the 24 hydroxycholesterol is converted into 7alpha-hydroxylated bile acids by the enzymes involved in the normal biosynthesis of bile acids. PMID- 11013307 TI - Distribution of phospholipid transfer protein in human plasma: presence of two forms of phospholipid transfer protein, one catalytically active and the other inactive. AB - Plasma phospholipid transfer protein (PLTP) plays an important role in the maintenance of plasma high-density lipoprotein (HDL) content and remodeling of HDL in the circulation. In the present study we have used different fractionation methods to investigate the distribution of PLTP in human plasma. A novel enzyme linked immunosorbent assay developed during the study allowed for simultaneous assessment of both PLTP mass and activity in the fractions obtained. Size exclusion chromatography and plasma fractionation by nondenaturing polyacrylamide gel electrophoresis (PAGE) yielded similar results demonstrating that PLTP associates in native plasma with two distinct particle populations, while ultracentrifugation with high salt leads to detachment of PLTP from lipoprotein particles and loss of a majority of its phospholipid transfer activity. Interestingly, analysis of the size-exclusion chromatography fractions demonstrated that PLTP exists in the circulation as an active population that elutes in the position of HDL corresponding to an average molecular mass of 160+/ 40 kDa and an inactive form with an average mass of 520+/-120 kDa. The inactive fraction containing approximately 70% of the total PLTP protein eluted between HDL and low density lipoprotein (LDL). Thus, the two PLTP pools are associated with different types of lipoprotein particles, suggesting that the PLTP activity in circulation is modulated by the plasma lipoprotein profile and lipid composition. PMID- 11013308 TI - Cholesterol delivered to macrophages by oxidized low density lipoprotein is sequestered in lysosomes and fails to efflux normally. AB - Oxidized low density lipoprotein (LDL) has been found to exhibit numerous potentially atherogenic properties, including transformation of macrophages to foam cells. It is believed that high density lipoprotein (HDL) protects against atherosclerosis by removing excess cholesterol from cells of the artery wall, thereby retarding lipid accumulation by macrophages. In the present study, the relative rates of HDL-mediated cholesterol efflux were measured in murine resident peritoneal macrophages that had been loaded with acetylated LDL or oxidized LDL. Total cholesterol content of macrophages incubated for 24 h with either oxidized LDL or acetylated LDL was increased by 3-fold. However, there was no release of cholesterol to HDL from cells loaded with oxidized LDL under conditions in which cells loaded with acetylated LDL released about one-third of their total cholesterol to HDL. Even mild degrees of oxidation were associated with impairment of cholesterol efflux. Macrophages incubated with vortex aggregated LDL also displayed impaired cholesterol efflux, but aggregation could not account for the entire effect of oxidized LDL. Resistance of apolipoprotein B (apoB) in oxidized LDL to lysosomal hydrolases and inactivation of hydrolases by aldehydes in oxidized LDL were also implicated. The subcellular distribution of cholesterol in oxidized LDL-loaded cells and acetylated LDL-loaded cells was investigated by density gradient fractionation, and this indicated that cholesterol derived from oxidized LDL accumulates within lysosomes. Thus impairment of cholesterol efflux in oxidized LDL-loaded macrophages appears to be due to lysosomal accumulation of oxidized LDL rather than to impaired transport of cholesterol from a cytosolic compartment to the plasma membrane. PMID- 11013309 TI - Increased low density lipoprotein degradation in aorta of irradiated mice is inhibited by preenrichment of low density lipoprotein with alpha-tocopherol. AB - We previously reported that upper thoracic exposure to ionizing radiation (IR) accelerates fatty streak formation in C57BL/6 mice and that such effects are inhibited by overexpression of the antioxidant enzyme CuZn-superoxide dismutase (SOD). Notably, IR-accelerated lesion formation is strictly dependent on a high fat diet (i.e., atherogenic lipoproteins) but does not involve alterations in circulating lipid or lipoprotein levels. We thus proposed that IR promotes changes in the artery wall that enhance the deposition of lipoprotein lipids. To address this hypothesis, we examined the effects of IR on aortic accumulation and degradation of low density lipoproteins (LDL). Ten-week-old C57BL/6 mice were exposed to a single (8-Gy) dose of (60)Co radiation to the upper thoracic area or were sham irradiated (controls) and were then placed on the high fat diet. Five days postexposure, the mice received either (125)I-labeled LDL ((125)I-LDL) (which was used to measure intact LDL) or (125)I-labeled tyramine cellobiose ((125)I-TC)-LDL (which was used to measure both intact and cell-degraded LDL) via tail vein injection. On the basis of trichloroacetic acid (TCA)-precipitable counts in retroorbital blood samples, > or =95% of donor LDL was cleared within 24 h and there were no differences in time-averaged plasma concentrations of the two forms of LDL among irradiated and control mice. Aortic values increased markedly within the first hour and thereafter exhibited a slow increase up to 24 h. There were no differences between irradiated and control mice at 1 h, when values primarily reflected LDL entry, but a divergence was observed thereafter. At 24 h, (125)I-TC-associated counts were 1.8-fold higher in irradiated mice (P = 0.10). In contrast, (125)I-LDL-associated counts were 30% lower in irradiated mice (P< 0.05), suggesting that most of the retained (125)I-TC was associated with LDL degradation products. Consistent with the proposed involvement of oxidative or redox-regulated events, IR-induced LDL degradation was lower in SOD transgenic than wild-type mice (P<0.05). The importance of LDL oxidation was suggested by observations that IR-induced LDL degradation was significantly reduced by preenriching LDL with alpha-tocopherol. On the basis of these results, we propose that IR elicits SOD-inhibitable changes in the artery wall that enhance LDL oxidation and degradation leading to the deposition of LDL-borne lipids. These studies provide additional support for the role of oxidation in lipoprotein lipid deposition and atherogenesis and suggest that IR promotes an arterial environment that stimulates this process in vivo. PMID- 11013310 TI - Hepatic apolipoprotein E expression promotes very low density lipoprotein apolipoprotein B production in vivo in mice. AB - In addition to its role in the uptake of apolipoprotein B (apoB)-containing lipoproteins, apoE promotes hepatic very low density lipoprotein-triglyceride (VLDL-TG) production in animal models. However, it is not known if apoE increases the amount of TG per VLDL particle or the number of VLDL particles secreted. VLDL apoB production is a measure of the rate of VLDL particle secretion. We determined the effects of apoE deficiency and apoE overexpression on VLDL-apoB production in mice. [(35)S]methionine was injected into endogenously label VLDL apoB and Triton WR-1339 was simultaneously injected to block the catabolism of VLDL. Compared with wild-type mice, the VLDL-apoB production rate was decreased by 33% in apoE-deficient mice. Conversely, VLDL-apoB production was increased by 48% in mice overexpressing apoE compared with controls. Nascent VLDL, obtained from post-Triton plasma, had a decreased, not increased, content of TG per apoB in the apoE-overexpressing group compared with the control group. This study demonstrates that hepatic apoE expression increases the output of VLDL triglyceride by increasing the production rate of VLDL-apoB, suggesting that hepatic apoE influences the number of VLDL particles secreted by the liver. PMID- 11013311 TI - Sphingolipids and cholesterol modulate membrane susceptibility to cytosolic phospholipase A(2). AB - Modulation of cytosolic phospholipase A(2) (cPLA(2)) activity by sphingomyelin (SPH), ceramide (Cer), and cholesterol (Chol) was investigated in CHO-2B cells activated by the calcium ionophore A23187 and epinephrine. Chol depletion of CHO 2B cells by treatment with methyl-beta-cyclodextrin (5 mm) resulted in the inhibition of the release of arachidonic acid whereas the restoration of the level by Chol-loaded cyclodextrin relieved inhibition. Conversion of CHO-2B cellular SPH to Cer by Staphylococcus aureus sphingomyelinase enhanced endogenous cPLA(2) activation as well as uptake by cells of C2- and C6-ceramide analogs. These results were confirmed in vitro with purified human recombinant cPLA(2) acting on a model phospholipid substrate. The enzyme activity was inhibited by SPH but reactivated by Cer as well as by Chol added to glycerophospholipid liposomal substrates containing SPH. The results of this study, which combine in situ and in vivo experimental approaches, indicate that membrane microdomains enriched in SPH and Chol play a role in the modulation of the activity of cPLA2 and in arachidonic acid-derived mediator production. PMID- 11013312 TI - Eicosanoids as endogenous regulators of leptin release and lipolysis by mouse adipose tissue in primary culture. AB - Prostaglandin E(2) (PGE(2)) stimulated leptin release over a 24-h incubation of mouse adipose tissue in primary culture. The maximal stimulation of leptin release was seen with 100 nm PGE(2). The role of endogenous eicosanoids in the regulation of lipolysis and leptin formation was examined in the presence of NS 398, a selective cyclooxygenase-2 inhibitor. NS-398 at a concentration of 5 microm enhanced lipolysis by 30% and lowered leptin release by 24%. This concentration of NS-398 almost completely inhibited PGE(2) formation. An inhibition of basal lipolysis by PGE(2) or N(6)-cyclopentyladenosine (CPA) was seen in the presence but not in the absence of NS-398. CPA, whose receptor, like that of PGE(2) inhibits cyclic AMP accumulation in adipose tissue, also enhanced leptin release. These data indicate that PGE2 can stimulate leptin release and suggest that endogenous eicosanoids affect both lipolysis and leptin formation by mouse adipose tissue. PMID- 11013313 TI - International symposium on basic aspects of HDL metabolism and disease prevention. PMID- 11013314 TI - Taking issue: pharmacological progress: seeking balance. PMID- 11013316 TI - Datapoints: Effects of changing from five to ten preauthorized outpatient sessions. PMID- 11013317 TI - Law & psychiatry: Pegram v. Herdrich: the Supreme Court passes the buck on managed care. PMID- 11013318 TI - Personal accounts. Managed care wars: a first casualty. PMID- 11013319 TI - Managed care: Public-sector managed behavioral health care: VI. The Iowa approach to profit and community reinvestment. PMID- 11013321 TI - Economic grand rounds: Prevalence and cost of treating mental disorders among elderly recipients of Medicare services. PMID- 11013322 TI - Clozapine treatment of outpatients with schizophrenia: outcome and long-term response patterns. 1993. PMID- 11013323 TI - Drugs of the psychopharmacological revolution in clinical psychiatry. PMID- 11013325 TI - Client-case manager racial matching in a program for homeless persons with serious mental illness. AB - OBJECTIVE: This study evaluated the relationship between client-case manager racial matching and both service use and clinical outcomes in a case management program for homeless persons with serious mental illness. METHODS: The study focused on 1,785 clients from the first cohorts that entered the Center for Mental Health Services' Access to Community Care and Effective Services and Supports (ACCESS) program, a five-year demonstration program for homeless persons with mental illness established at 18 sites between 1994 and 1996. A series of two-way analyses of variance was used to assess the effect of client and case manager race and their interaction on changes in outcomes and service use over a 12-month period. RESULTS: Although African Americans had more severe problems on several measures and higher levels of service use at baseline, no differences in service use at 12 months or in the changes in client outcomes as measured by nine variables were associated with the different pairings of African-American and white clients and case managers. White clients had a greater reduction in psychotic symptoms than did African-American clients, regardless of client- case manager racial pairing. No differences were found between white and African American clients on the amount of services received over time. CONCLUSIONS: This study found virtually no evidence of a relationship between client race, case manager race, or client-case manager racial matching on either outcomes or service use. PMID- 11013324 TI - Racial differences in the prevalence of dementia among patients admitted to nursing homes. AB - OBJECTIVE: The purpose of this study was to determine the prevalence of dementia among black and white residents on admission to nursing homes and to determine whether demographic and health characteristics known to be associated with dementia were correlated with dementia in this population. METHODS: Data from medical records and structured interviews with family members, nursing staff, and nursing home residents were gathered for 2,285 persons newly admitted to nursing homes in Maryland from 1992 to 1995. A stratified sample of 59 nursing homes was used. An expert panel of five physicians classified each resident as demented, nondemented, or indeterminate. Associations between dementia status, race, and selected characteristics were examined. RESULTS: Black residents (77 percent) were significantly more likely than white residents (57 percent) to be classified as demented. Older age was associated with dementia in both races. Less education, male gender, and a history of a cerebrovascular accident were associated with an increased prevalence of dementia among white residents only. After demographic and health characteristics associated with dementia were controlled for, black race remained independently associated with a diagnosis of dementia. CONCLUSIONS: The rate of dementia on admission to nursing homes was higher among black residents than among white residents, a finding that has implications for the delivery of care. The higher rate may be due to psychosocial factors operating differently in blacks and whites that influence the timing of admission to a nursing home. PMID- 11013326 TI - A managed behavioral health organization operated by an academic psychiatry department. AB - As a means of adapting to managed care, the psychiatry department at Wake Forest University developed a managed behavioral health organization (MBHO) to manage the care of enrollees in QualChoice, the health maintenance organization of the Wake Forest University Baptist Medical Center. Before the academic MBHO was created, care was managed by a for-profit MBHO. In this case study, financial and utilization data were obtained from both MBHOs and from QualChoice. The data confirm that the academic MBHO was able to offer competitive rates for its services. It also was able to increase enrollees' use of the medical center's own providers and facilities by making more referrals than were made by the for profit MBHO. Developing a managed behavioral health organization can allow academic psychiatry departments, either individually or as consortia, to preserve the patient base they require for teaching, research, and financial stability. PMID- 11013327 TI - Admissions, length of stay, and medication use among women in an acute care state psychiatric facility. AB - OBJECTIVE: Despite growing numbers of elderly persons with serious mental illness such as schizophrenia, little research has been conducted on the manifestation of serious mental illness in later life, and our understanding of the mental health care needs of this population is limited. This study examined length of stay and medication use among women age 50 and older admitted to an acute care state psychiatric facility. METHODS: A computerized record search for all women discharged from a large urban state psychiatric facility over a one-year period was undertaken. Demographic and resource utilization data, including total length of stay and pharmacy utilization, were obtained from the hospital database. RESULTS: The database search produced the records of 564 women, with a mean age of 37.5 years. Women constituted 60 percent of patients over age 50. The mean length of stay was 16.1 days for the entire group; for women under age 50 (N=492), mean length of stay was 15 days, and for women age 50 and older (N=72), it was 23.1 days (p=. 01). Among those age 50 and older, 58.3 percent had a diagnosis of schizophrenia or schizoaffective disorder, compared with only 38 percent of those under age 50. CONCLUSIONS: Our data suggest that women constitute the majority of patients over age 50 in a state psychiatric facility and that they have longer stays than younger women. PMID- 11013328 TI - Bed closures and incarceration rates among users of Veterans Affairs mental health services. AB - OBJECTIVE: This study examined incarceration rates of users of Department of Veterans Affairs (VA) mental health services in 16 northeastern New York State counties between 1994 and 1997-a time of extensive bed closures in the VA system to determine whether incarceration rates changed during this period. METHODS: Data were obtained for male patients who used inpatient and outpatient VA mental health services between 1994 and 1997 and for men incarcerated in local jails during this period. For comparison, services use and incarceration data were obtained for all men who received inpatient behavioral health care at community general hospitals and state mental hospitals between 1994 and 1996 in the same counties. Probabilistic population estimation, a novel statistical technique, was employed to evaluate the degree of overlap between clinical and incarceration populations without relying on person-specific identifiers. RESULTS: Of all male users of VA mental health services between 1994 and 1997, a total of 15.7 percent 39.6 percent of those age 18 to 39 years and 9.1 percent of those age 40 years and older-were incarcerated at some time during that period. Dual diagnosis patients had the highest rate of incarceration (25 percent), followed by patients with substance abuse problems only (21 percent) and those with mental health problems only (11 percent). The rate of incarceration among male patients hospitalized in VA facilities was lower than among men in general hospitals or state hospitals (11.6 percent, 23 percent, and 21.7 percent, respectively), but was not significantly different. No significant increase occurred in the annual rate of incarceration among VA patients from 1994 to 1997 (3.7 percent to 4 percent), despite extensive VA bed closures during these years. CONCLUSIONS: Substantial proportions of mental health system users were incarcerated during the study period, especially younger men and those with both substance use and mental health disorders. Rates of incarceration were similar across health care systems. The closure of a substantial number of VA mental health inpatient beds did not seem to affect the rate of incarceration among VA service users. PMID- 11013329 TI - The relationship between command hallucinations and violence. AB - OBJECTIVE: The purpose of this study was to describe the relationship between command hallucinations and violent behavior. METHODS: One hundred and three psychiatric inpatients completed measures of command hallucinations, other psychotic symptoms, violent behavior, and social desirability response biases. RESULTS: Thirty percent of the patients reported having had command hallucinations to harm others during the last year, and 22 percent of the patients reported they complied with such commands. Logistic regression analyses suggested that patients who experienced command hallucinations to harm others were more than twice as likely to be violent, even when the analysis controlled for demographic variables, history of substance abuse, and social desirability response biases. CONCLUSIONS: The results support the clinical utility of asking about command hallucinations when assessing the risk of violence in patients with major mental disorders. PMID- 11013330 TI - Help seeking by persons of Mexican origin with functional impairments. AB - OBJECTIVE: Mexican Americans have low rates of service utilization for mental health problems. This study examined the use of health services by persons who have symptoms of mental distress that they say impair their ability to function. METHODS: A stratified field survey was conducted in central California to select a probabilistic sample of persons of Mexican origin. A multinomial logistic regression was used for the analysis. RESULTS: Among respondents who reported functional impairments attributable to their symptoms, 58 percent met diagnostic criteria for a mental disorder. Of that group, 69 percent did not use any health services. Respondents who sought medical services for mental health symptoms were more likely to be women and to meet diagnostic criteria for a mental disorder. Respondents who sought mental health services for their symptoms were more likely to be female, to be unmarried, and to have had 12 or more years of education. Health insurance status was not associated with type of service used. CONCLUSIONS: Even among Mexican Americans who met diagnostic criteria for a mental illness, most did not use services of any sort, and many of those who sought treatment for their symptoms turned to medical and informal services. PMID- 11013331 TI - Principal and additional DSM-IV disorders for which outpatients seek treatment. AB - OBJECTIVE: Epidemiological studies indicate that most patients in the community do not get treatment for psychiatric disorders. It is unknown whether persons who present for outpatient psychiatric services seek treatment for all the disorders they have or only for the principal disorder for which they are seeking treatment. The goal of this study was to determine which axis I psychiatric disorders motivate patients to seek treatment. METHODS: Four hundred outpatients at a hospital-affiliated, community-based, psychiatric clinical practice were interviewed with the Structured Clinical Interview for DSM-IV (SCID). For patients with more than one disorder, the diagnoses were assigned as principal or additional according to the DSM-IV convention of whether it was the patient's stated primary reason for presenting for treatment or was an additional disorder. For all current disorders, patients were asked whether the symptoms of each diagnosed disorder were a reason, or one of the reasons, for seeking treatment. RESULTS: Nearly all patients with major depression wanted treatment for this disorder, and more than 85 percent of patients with panic disorder, posttraumatic stress disorder, and generalized anxiety disorder indicated that the symptoms of these disorders were a reason for seeking treatment. Half to two-thirds of patients with social phobia, obsessive-compulsive disorder, intermittent explosive disorder, body dysmorphic disorder, and substance use disorders reported that the symptoms of these disorders were a reason for seeking treatment. Only 30 percent of those with specific phobia indicated that their phobic fears were a reason for seeking treatment. CONCLUSIONS: Patients often seek treatment for symptoms of disorders that are diagnosed as comorbid, rather than principal, conditions. It is important for clinicians to conduct thorough diagnostic interviews in order to diagnose disorders that are not related to the patient's chief complaint, as patients often desire treatment for these additional diagnoses. PMID- 11013332 TI - Satisfaction of forensic psychiatric patients with remote telepsychiatric evaluation. AB - The level of satisfaction with telepsychiatry evaluations was determined in a sample of 43 forensic psychiatric patient inmates in a large urban jail. A forensic psychiatrist interviewed 20 patients in person, the other 23 remotely via interactive video. Demographic characteristics, physical health status, and psychiatric symptom severity on the Global Severity Index of the Brief Symptom Inventory were comparable in the two groups. Patient satisfaction with the evaluations was moderately high for patients in both groups, with no significant differences between them. PMID- 11013333 TI - Establishing a function-based mental health service line in a VA medical center. AB - From 1994 through 1996, a general Veterans Affairs (VA) medical center reorganized its mental health services from a traditional discipline-based structure to a unitary service line organized around patient care functions. A comparison of data from 1993 and 1997 indicated increased efficiency, substantial transfer of patients from inpatient to outpatient care, and growth in academic programs not explainable solely by temporal, regional, or national trends or by trends within the VA medical center. Although the results should be interpreted conservatively because of the observational nature of the study, the reorganization appeared to facilitate the positive changes that occurred over the study period. PMID- 11013334 TI - Coping resources of African-American and white patients hospitalized for bipolar disorder. AB - Forty-two African-American and 80 white patients hospitalized for bipolar disorder completed the Coping Resources Inventory. Total resource scores of the African Americans were significantly higher than scores of the whites, although differences in background variables between the two groups were not evident. The African-American group also scored significantly higher than the whites on the three scales indicating internal resources-cognitive, emotional, and spiritual philosophical. No statistically significant differences were found for the two groups on the social and physical scales. Cultural orientations influencing personal life philosophies may explain the differences between the African American and white patients on perceptions of their coping resources. PMID- 11013335 TI - Identifying and using reinforcers to enhance the treatment of persons with serious mental illness. AB - Successful engagement of clients with serious and persistent mental illnesses in the planning and implementation of treatment requires the identification of individualized reinforcers through motivational analyses. Reinforcer surveys are interviews or questionnaires that list numerous objects, persons, activities, and settings, and then assess the client's perception of the value of each item. Such surveys help identify the type of reward that might be useful for motivating the client. If properly assessed and delivered, reinforcers can increase clients' skill acquisition, attainment of goals, and feelings of self-efficacy. PMID- 11013336 TI - Improving police response to mentally ill people. PMID- 11013337 TI - Linking discharged patients with peers in the community. PMID- 11013338 TI - "I vote. I count.". PMID- 11013339 TI - Tarasoff warnings. PMID- 11013340 TI - Seclusion and restraints. PMID- 11013342 TI - 1alpha-Hydroxylase and the action of vitamin D. AB - The active form of vitamin D, 1,25-dihydroxvitamin D(3) (1, 25(OH)(2)D(3)), is a pleiotropic hormone whose actions include the regulation of calcium homeostasis, control of bone cell differentiation and modification of immune responses. Synthesis of 1, 25(OH)(2)D(3) from the major circulating metabolite, 25 hydroxyvitamin D(3) (25(OH)D(3)), is catalysed by a mitochondrial cytochrome P450 enzyme, 25-hydroxyvitamin D-1alpha-hydroxylase (1alpha-OHase). Although 1alpha OHase is expressed predominantly in the kidney, extra-renal production of 1,25(OH)(2)D(3) has also been demonstrated in tissues such as lymph nodes and skin. The tight regulation of 1alpha-OHase which occurs in both renal and peripheral tissues has made studies of the expression and regulation of this enzyme remarkably difficult. However, the recent cloning of mouse, rat and human cDNAs for 1alpha-OHase (CYP1alpha/Cyp1alpha) has enabled a more thorough characterization of this enzyme. In particular, analysis of the CYP1alpha gene has identified mutations causing the inherited disorder vitamin D-dependent rickets type 1, also known as pseudo-vitamin D deficiency rickets. Studies from our own group have focused on the distribution of 1alpha-OHase in both renal and extra-renal tissues. Data indicate that the enzyme is expressed throughout the nephron, suggesting discrete endocrine and paracrine/autocrine functions. Further immunohistochemical analyses have shown that the enzyme is widely distributed in extra-renal tissues, and this appears to be due to the same gene product as the kidney. Collectively, these observations have raised important new questions concerning the role of 1alpha-OHase in vitamin D signalling at a local level. The relationship between expression of protein for 1alpha-OHase and enzyme activity has yet to be fully characterized and may be dependent on membrane proteins such as megalin. Similarly, elucidation of the mechanisms involved in differential regulation of renal and extra-renal 1,25(OH)(2)D(3) production will be essential to our understanding of the tissue-specific functions of 1alpha-OHase. These and other issues are discussed in the current review. PMID- 11013343 TI - Role of aromatase in sex steroid action. PMID- 11013344 TI - Molecular evolution of the growth hormone-releasing hormone/pituitary adenylate cyclase-activating polypeptide gene family. Functional implication in the regulation of growth hormone secretion. AB - Growth hormone-releasing hormone (GHRH) and pituitary adenylate cyclase activating polypeptide (PACAP) belong to the same superfamily of regulatory neuropeptides and have both been characterized on the basis of their hypophysiotropic activities. This review describes the molecular evolution of the GHRH/PACAP gene family from urochordates to mammals and presents the hypothesis that the respective roles of GHRH and PACAP in the control of GH secretion are totally inverted in phylogenetically distant groups of vertebrates. In mammals, GHRH and PACAP originate from distinct precursors whereas, in all submammalian taxa investigated so far, including birds, amphibians and fish, a single precursor encompasses a GHRH-like peptide and PACAP. In mammals, GHRH-containing neurons are confined to the infundibular and dorsomedial nuclei of the hypothalamus while PACAP-producing neurons are widely distributed in hypothalamic and extrahypothalamic areas. In fish, both GHRH- and PACAP-immunoreactive neurons are restricted to the diencephalon and directly innervate the adenohypophysis. In mammals and birds, GHRH plays a predominant role in the control of GH secretion. In amphibians, both GHRH and PACAP are potent stimulators of GH release. In fish, PACAP strongly activates GH release whereas GHRH has little or no effect on GH secretion. The GHRH/PACAP family of peptides thus provides a unique model in which to investigate the structural and functional facets of evolution. PMID- 11013345 TI - Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. AB - The reverse transcription polymerase chain reaction (RT-PCR) is the most sensitive method for the detection of low-abundance mRNA, often obtained from limited tissue samples. However, it is a complex technique, there are substantial problems associated with its true sensitivity, reproducibility and specificity and, as a quantitative method, it suffers from the problems inherent in PCR. The recent introduction of fluorescence-based kinetic RT-PCR procedures significantly simplifies the process of producing reproducible quantification of mRNAs and promises to overcome these limitations. Nevertheless, their successful application depends on a clear understanding of the practical problems, and careful experimental design, application and validation remain essential for accurate quantitative measurements of transcription. This review discusses the technical aspects involved, contrasts conventional and kinetic RT-PCR methods for quantitating gene expression and compares the different kinetic RT-PCR systems. It illustrates the usefulness of these assays by demonstrating the significantly different levels of transcription between individuals of the housekeeping gene family, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). PMID- 11013346 TI - Identification of a novel calcitonin-response element in the promoter of the human p21WAF1/CIP1 gene. AB - The cyclin-dependent kinase inhibitor p21/WAF1/CIP1 is induced in many cell types in response to a variety of extracellular signals and causes cell cycle arrest in both the G1 and G2/M phases of the cell cycle. We reported previously that calcitonin (CT) receptor (CTR)-mediated growth inhibition of HEK-293 cells stably transfected with the human CTR is accompanied by a rapid and sustained induction of p21 and cell cycle arrest in G2. In the present study we have shown that CT stimulates transcription from a p21 promoter-luciferase construct. Using deletion and mutation analysis of the p21 promoter we have demonstrated that transcriptional activation of p21 by CT is p53-independent and is mediated through specific activation of Sp1-binding sites in a region of the promoter between -82 and -69, relative to the transcription start site. CTR-mediated transcriptional activation of p21 was specific for the insert-negative isoform of the human CTR. Butyrate, which was shown previously to activate the same Sp1 site, synergised with CT to increase further p21 promoter activity. These results provide the first demonstration that CTR can induce gene transcription through the constitutively expressed transcription factor Sp1, and define a mechanism of cell growth suppression that may have implications for other members of the seven transmembrane domain G protein-coupled receptor superfamily. PMID- 11013347 TI - Oestradiol decreases rat apolipoprotein AI transcription via promoter site B. AB - Oestrogens protect against ischaemic heart disease in the post-menopausal female by increasing serum concentrations of apolipoprotein (apo) AI and the abundance of high-density lipoprotein particles. In men and experimental male animals, the administration of oestrogen has variable effects on apo AI expression. As the major mode of oestrogen action on target genes involves regulating promoter activity and hence transcription, oestrogen is expected to alter transcription of the apo AI gene. To test this hypothesis, the effect of 17beta-oestradiol (E(2)), on rat apo AI promoter activity in male hepatoma HuH-7 cells, was tested by co transfecting a reporter template, pAI.474.CAT containing-474 to-7 of the rat apo AI promoter and an oestrogen receptor (ER) expression vector, pCMV-ER. Transfected cells exposed to E(2) showed a dose-dependent decrease in chloramphenicol acetyltransferase (CAT)-activity, with a maximum 91+/-1.5% reduction at 1 microM E(2). Deletional analysis of the promoter localized the inhibitory effect of ER and E(2) to site B (-170 to-144) with an adjacent 5' contiguous motif, site S (-186 to-171) acting as an amplifier. HuH-7 cell nuclear extracts showed binding activities with both sites S and B, but recombinant human ER did not. Furthermore, nuclear extracts from E(2)-treated HuH-7 cells showed weaker binding activity to site B, but not to site S. In summary, the inhibitory effect of ER and E(2) on rat apo AI gene activity is mediated by a promoter element, site B. This inhibitory effect arises from a mechanism that does not involve direct ER binding to the B-element. The conclusion that E(2) inhibits apo AI transcription was confirmed in vivo. Treatment of male adult Sprague-Dawley rats with up to 200 microg E(2) for 7 days decreased apo AI protein and hepatic mRNA by 72+/-21% and 68+/-1.4% respectively. Results of 'run-on' transcription of the apo AI gene in isolated hepatic nuclei showed a 55% decrease in hormone treated male rats. These findings suggest that E(2) exerts primarily an inhibitory effect within male hepatic nuclei. PMID- 11013348 TI - Characterization of a human 20alpha-hydroxysteroid dehydrogenase. AB - It has been suggested that 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) is a T-cell differentiation marker in mice. In the human, this enzyme has generally been associated with types 1 and 2 17beta-HSDs, which belong to the short-chain alcohol dehydrogenase family, whereas the rat, rabbit, pig and bovine 20alpha HSDs are members of the aldoketo reductase superfamily, which also includes the 3alpha-HSD family. In this study, we report the cloning, from a human skin cDNA library, of a cDNA that shows, after transfection into human embryonic kidney (HEK-293) cells, high 20alpha-HSD activity but negligible 3alpha- and 17beta hydroxysteroid dehydrogenase activities. A comparison of the amino acid sequence of the human 20alpha-HSD with those of other related 20alpha- and 3alpha-HSDs indicates that the human 20alpha-HSD shares 79.9, 68.7 and 52.3% identity with rabbit, rat and bovine 20alpha-HSDs, whereas it shows 97, 84 and 65% identity with human type 3, type 1 and rat 3alpha-HSDs. In contrast, the enzyme shares only 15.2 and 15.0% identity with type 1 and type 2 human 17beta-HSDs. DNA analysis predicts a protein of 323 amino acids, with a calculated molecular weight of 36 767 Da. In intact transfected cells, the human 20alpha-HSD preferentially catalyzes the reduction of progesterone to 20alpha hydroxyprogesterone with a K(m) value of 0.6 microM, the reverse reaction (oxidation) being negligible. In a cell cytosolic preparation, the enzyme could use both NADPH and NADH as cofactors, but NADPH, which gave 4-fold lower K(m) values, was preferred. We detected the expression of 20alpha-HSD mRNA in liver, prostate, testis, adrenal, brain, uterus and mammary-gland tissues and in human keratinocyte (HaCaT) cells. The present study clearly indicates that the genuine human 20alpha-HSD belongs to the aldoketo reductase family, like the 20alpha-HSDs from other species. PMID- 11013349 TI - Functional characteristics of a novel murine estrogen receptor-beta isoform, estrogen receptor-beta 2. AB - We have isolated a highly expressed splice variant mRNA of murine estrogen receptor-beta (ERbeta), mERbeta2, containing an in-frame 54 nucleotide insertion between exons 5 and 6 of wild-type mERbeta1. The predicted ERbeta2 protein contains 18 amino acids inserted in the ligand binding domain of mERbeta1. Recombinant protein generated by in vitro transcription/translation showed that mERbeta2 had markedly reduced ligand binding (K(D)=17.7+/-4.7 nM, mean+/-s.e.m., n=3) compared with mERbeta1-bound (3)H-estradiol (K(D)=0.56+/- 0.19 nM, mean+/ s.e.m., n=3). Both receptors bound similarly to palindromic estrogen responsive elements (EREs) in vitro and in vivo, and similarly bent DNA. Transcriptional activity was assessed using transient transfection analysis into a homologous murine cell line, NIH 3T3 cells. mERbeta1 transactivated ERE-tk-CAT reporter genes similarly to mERalpha, whereas mERbeta2 had little activity except at high ligand concentrations. However, under conditions in which mERbeta2 is unlikely to be ligand saturated, co-transfected mERbeta2 inhibited activity of mERalpha and possibly mERbeta1 on ERE-tk-CAT genes. Using a 'novel raloxifene responsive' gene reporter system (TGF-beta3-CAT), we found the ability of estradiol and LY117018 to activate both mERalpha and mERbeta1 on this promoter was identical, and mERbeta2 activity in the presence of either estradiol or LY117018 was only slightly less than that observed with either mERbeta1 or mERalpha. Both mERbeta1 and mERbeta2 when liganded with LY117018 inhibited transcription at a classical ERE-regulated promoter under these transfection conditions, which was in marked contrast to their stimulatory effect at the transforming growth factor-beta3 promoter. These data suggest that responsiveness of gene expression to a relatively highly expressed variant murine ERbeta isoform, mERbeta2, is both ligand and promoter specific. Determination of the relative level of expression of mERbeta1 mRNA and mERbeta2 mRNA in mouse tissues indicated predominance of mERbeta2 mRNA in some but not all tissues. These data suggest that the mERbeta2 may have some tissue-specific and promoter-specific modulatory effects. PMID- 11013350 TI - Quantification of rainbow trout (Oncorhynchus mykiss) estrogen receptor-alpha messenger RNA and its expression in the ovary during the reproductive cycle. AB - This study developed a quantitative reverse transcription-polymerase chain reaction (RT-PCR) method to measure estrogen receptor-alpha (ERalpha) mRNA in the rainbow trout (Oncorhynchus mykiss). Using RT-PCR, and primers based on the known ERalpha DNA sequence in this species, cDNA sequences representing most of the protein coding region were obtained from ovary poly A(+) RNA. Using these DNA sequences as probes in Northern blot hybridizations confirmed that a single transcript of 4.2 kilobases in poly A(+) RNA could be detected in liver and ovary RNA. For the quantitative RT-PCR assay an internal standard RNA molecule was produced to control for inherent inter-tube differences in amplification efficiency and permit accurate quantification of ERalpha mRNAs. The quantitative RT-PCR assay proved to be highly specific for ERalpha mRNA with a detection limit of 6.9 fg, which corresponds to 273 fg ERalpha mRNA/microg total RNA. The quantitative RT-PCR assay was used to measure the levels of ERalpha mRNA in ovaries of rainbow trout at different stages of reproductive development. Ovarian ERalpha mRNA expression was found during two distinct periods of reproductive development, in pre-vitellogenic ovaries of fish with ovarian follicle diameters (OFDs) 1000 microm. ERalpha mRNA could not be detected in the ovaries of fish with OFDs >100 microm but 2000 microm. PMID- 11013351 TI - Induction of erythroid proliferation and differentiation by a trophoblast specific cytokine involves activation of the JAK/STAT pathway. AB - Pregnancy is characterized by increased erythropoiesis within maternal and fetal compartments. The placenta has been shown to produce factors that stimulate erythropoiesis but convincing evidence for placental production of erythropoietin (EPO) is still lacking. Prolactin-like protein E (PLP-E) was recently found to stimulate expression of the adult beta major globin gene in mouse erythroleukemia cells. Here we demonstrate that PLP-E transiently expressed in COS-7 cells stimulates proliferation and erythroid differentiation of murine and human erythroid progenitor cell lines. Electrophoretic mobility shift assays were used to show the activation of STAT5 by PLP-E in the human erythroid cell line TF1. Furthermore, we compared the effects of PLP-E on murine myeloid FDCP1 cells which do not express EPO receptors (EPORs) with effects on cells genetically engineered to express functional EPORs. We provide evidence that PLP-E-dependent proliferation and STAT5 activation is independent of the expression of the EPOR. Taken together, these data suggest that PLP-E acts on specific receptors of erythroid-committed murine and human cells by the activation of intracellular signaling pathways promoting cell growth and differentiation. PMID- 11013352 TI - CYFRA 21-1 determination in patients with esophageal squamous cell carcinoma: clinical utility for detection of recurrences. AB - BACKGROUND: While there are reports that CYFRA 21-1 is a useful tumor marker, to our knowledge the clinical utility of this marker to detect recurrences for squamous cell carcinoma of the esophagus has not been addressed. METHODS: By immunoradiometric assay, human serum levels of CYFRA 21-1, SCC antigen and CEA were measured in esophageal squamous cell carcinoma patients prior to their initial treatment. Monthly follow-ups of these tumor markers was done after surgery. RESULTS: The diagnostic sensitivity of CYFRA 21-1 was 43.9% (18 of 41), a value superior to that for SCC antigen (26.8%) and CEA (17.0%) (P < 0.05). The positive rates of CYFRA 21-1 increased with progression of the disease, 22.2% of pTNM Stage 0-IIA and 77.8% of pTNM Stage IIB/III (P = 0.013), whereas SCC antigen and CEA rates were not related to pTNM stage. Among 13 patients with clinical evidence of a recurrence, 76.9% (10 of 13) exhibited an increase in CYFRA 21-1, and this increase was evident before clinical detection of the recurrence in 9 of these 13 patients (69.2%). Consequently, postoperative elevations of serum CYFRA 21-1 levels were indicative of a tumor recurrence 1-13 months before acquisition of clinical and radiological data. CONCLUSIONS: The assay of CYFRA 21-1 is useful not only for diagnosis but also for close monitoring of patients with esophageal squamous cell carcinoma. PMID- 11013353 TI - Pathology and prognosis of gastric carcinoma: well versus poorly differentiated type. AB - BACKGROUND: The most important parameters predicting outcome of patients with gastric carcinoma are the depth of wall invasion and the status of lymph node metastasis, but the prognostic significance of histologic type is unclear. The aim of this study was to clarify the prognostic value of two major histologic types of gastric carcinoma, that is well and poorly differentiated types. METHODS: Histopathologic findings and outcomes of 504 patients with gastric carcinoma were evaluated by well and poorly differentiated types. Well differentiated gastric carcinoma (WGC) included papillary and tubular adenocarcinomas, poorly differentiated medullary carcinoma, and well differentiated mucinous carcinoma; whereas poorly differentiated gastric carcinoma (PGC) included poorly differentiated scirrhous carcinoma, signet ring cell carcinoma, and poorly differentiated mucinous carcinoma. RESULTS: Patients with WGC were characterized by old age, male predominance, tumor location in the lower third of the stomach, small tumor size, and liver metastasis; whereas patients with PGC were distinguished by their tumor location in the middle third of the stomach, serosal invasion, lymph node metastasis, advanced stage, and peritoneal dissemination. The overall 5-year survival rate for patients with WGC was higher than that for patients with PGC (76% vs. 67%; P = 0.058), especially for patients with >/= 10 cm tumors (42% vs. 14%; P = 0.017). The 5-year survival rate for patients with serosa positive but node negative tumors was higher in WGC patients than in PGC patients (83% vs. 59%; P = 0.086); whereas the 5-year survival rate for patients with serosa negative but node positive tumors was lower in WGC patients than in PGC patients (63% vs. 88%; P = 0.008). Multivariate analysis indicated that among pathologic variables of the tumor, histologic type (WGC vs. PGC) was one of the independent prognostic factors. CONCLUSIONS: Histologic type is important for estimating the tumor progression and outcomes of patients with gastric carcinoma. In addition to the depth of wall invasion and status of lymph node metastasis, histologic type, including well or poorly differentiated type, should be evaluated in the management of gastric cancer. PMID- 11013354 TI - Outcome of prophylactic radical lymphadenectomy with gastrectomy in patients with early gastric carcinoma without lymph node metastasis. AB - BACKGROUND: Prophylactic extended lymphadenectomy with gastrectomy may prolong survival in patients with early gastric carcinoma without lymph node metastasis. However, the therapeutic value of extensive lymphadenectomy in patients with early gastric carcinoma remains controversial. METHODS: The authors retrospectively analyzed 423 patients with early gastric carcinoma without lymph node metastasis who underwent gastrectomy and did not die of other diseases to evaluate the effect of prophylactic extended lymphadenectomy on postoperative survival. The postoperative survival rate of patients who underwent prophylactic extended lymphadenectomy was compared with that of patients who underwent prophylactic limited lymphadenectomy. RESULTS: Although extended lymphadenectomy did not appear to improve the postoperative survival rate of patients with mucosal tumors, it did improve the postoperative survival rate of patients with submucosal tumors. Whether prophylactic extended lymphadenectomy was performed significantly affected outcome in patients with early gastric carcinoma who had submucosal tumors without regional lymph node metastasis. CONCLUSIONS: Extensive lymphadenectomy with gastrectomy should be performed to prolong the survival of the patients with submucosal tumors. PMID- 11013355 TI - Detection of Helicobacter species in the liver of patients with and without primary liver carcinoma. AB - BACKGROUND: Several studies have shown the presence of Helicobacter species in the human biliary tract and in the intestinal tract of animals. In this study, the presence of Helicobacter species in liver samples from patients with primary hepatic carcinomas was evaluated. METHODS: Sixteen liver specimens were studied (8 from patients with primary liver carcinoma and 8 from patients without primary liver carcinoma). Histology with standard stains, culture, and polymerase chain reaction (PCR) amplification using two sets of primers located in the 16S ribosomal DNA (rDNA) were used to detect the presence of bacteria. Amplified products were sequenced to determine the genus and species of the bacteria. A search for other genes that were specific for Helicobacter pylori also was carried out by PCR. RESULTS: PCR performed with the 16S rDNA primers revealed the presence of bacteria from the genus Helicobacter in all of the liver specimens from patients with primary liver carcinoma (eight of eight patients) and in one specimen from a patient without primary liver carcinoma (one of eight patients). When the nucleotide sequence of > 80% of the 16S rDNA was determined, the closest similarity was with the 16S rDNA from H. pylori in eight patients. In 1 patient sample from which only 398 nucleotides were sequenced, the closest match was Helicobacter felis. CONCLUSIONS: The results presented in this study indicate that Helicobacter species can be present in the liver of patients with primary hepatic carcinoma, but their eventual role in the carcinogenesis process, although it is plausible, remains to be proven. Based on sequence similarity, it seems that Helicobacter species that are related closely to H. pylori but are distinct from it have been found. PMID- 11013356 TI - Glutathione depletion causes cell growth inhibition and enhanced apoptosis in pancreatic cancer cells. AB - BACKGROUND: Recent studies have demonstrated that various tumors express enhanced levels of the radical scavenger glutathione (GSH). Moreover, there are grounds for claiming that GSH plays a crucial role in cell proliferation and tumor resistance. In the current study, we investigated the relation between cell growth and GSH levels in the pancreatic adenocarcinoma cell line, AsPC-1, and the significance of GSH in tumor resistance to chemotherapy. METHODS: Cell growth in AsPC-1 was initiated through transforming growth factor-alpha (TGF-alpha) or fetal calf serum (FCS). Then, cell cycle, cell proliferation, and cellular GSH content were analyzed at different times in the presence or absence of buthionine sulfoximine (BSO). The impact of GSH on chemotherapy-induced apoptosis was studied using 5-fluorouracil or melphalan in the presence or absence of BSO. Finally, we compared the GSH content of 15 pancreatic tumor specimens with 10 normal pancreatic tissue specimens. RESULTS: Analysis of GSH in pancreatic tissues demonstrated increased GSH levels in cancerous compared with normal tissue (17.5 +/- 2.3 vs. 8. 8 +/- 1.4 nmol/mg protein; P < 0.004). Incubation of AsPC-1 with TGF-alpha or FCS resulted in cell proliferation and cell cycle activity, whereas GSH content was not altered. Incubation of GSH-depleted cells with TGF-alpha did not stimulate cell growth. In addition, GSH-depletion resulted in an increased rate of apoptosis after melphalan (6.3 +/- 0.3 % vs. 11.2 +/- 0.3 %; P < 0.001), but not after 5-fluorouracil treatment. CONCLUSIONS: Taken together, our results show enhanced GSH levels in pancreatic carcinoma and an essential role of GSH in cell proliferation and in resistance of AsPC-1 cells. Therefore, GSH-depletion may improve the efficacy of adjuvant therapy in pancreatic carcinoma. PMID- 11013357 TI - Increased secretory leukoprotease inhibitor in patients with nonsmall cell lung carcinoma. AB - BACKGROUND: It is well known that acute pulmonary inflammation, such as that observed in pneumonia, elevates secretory leukoprotease inhibitor (SLPI) levels in serum. A previous report indicated that serum SLPI levels in lung carcinoma patients with concurrent pneumonia were significantly higher than in those in patients whose disease was unaccompanied by pneumonia or in healthy subjects. The authors hypothesized that serum SLPI may increase in patients with lung carcinoma, even carcinoma occurring without pneumonia, and that cells in lung carcinoma might produce SLPI. METHODS: Serum SLPI levels in 58 patients with primary lung carcinoma unaccompanied by pneumonia and in 42 healthy subjects were measured by an enzyme immunoassay. Twenty-four specimens from 24 of the patients with primary lung carcinoma also were examined immunohistochemically using the rabbit antihuman SLPI antibody. RESULTS: The results of the current study confirmed that the serum levels of SLPI in patients with primary lung carcinoma were higher than those in healthy subjects, and further found there was no significant correlation between serum SLPI levels and C-reactive protein in lung carcinoma patients without pneumonia. When classifying primary lung carcinoma by its histology, SLPI levels in patients with adenocarcinoma and squamous cell carcinoma were significantly higher than in those in patients with small cell lung carcinoma (SCLC). In patients with nonsmall cell lung carcinoma (NSCLC), the SLPI levels in the advanced group (International Union Against Cancer Stages III and IV disease; n = 35) were significantly elevated compared with the nonadvanced group (Stages I and II disease; n = 12), and such elevated SLPI levels were reduced in some cases by an efficient response to surgical therapy or chemotherapy. Immunohistochemical studies showed that all the NSCLC tissues were stained with anti-human SLPI antibody, whereas staining was not noted in any of the SCLC tissues. CONCLUSIONS: The authors believe that the findings of the current study demonstrate that cells of NSCLC produce SLPI. Furthermore, they suggest that serum SLPI levels in serum may be a helpful marker in patients with NSCLC unaccompanied by pneumonia, and that SLPI also could be used as an immunohistochemical marker to distinguish between NSCLC and SCLC. PMID- 11013358 TI - Ki-67 immunostaining and other prognostic factors including tobacco smoking in patients with resected nonsmall cell lung carcinoma. AB - BACKGROUND: To estimate the effectiveness of expression of the tumor proliferative marker Ki-67 antigen (Ki-67) as a postoperative prognostic marker, the authors analyzed Ki-67 expression and its correlation with postoperative survival and other clinicopathologic factors, including preoperative smoking habits, in patients with resected nonsmall cell lung carcinoma (NSCLC). METHODS: A total of 156 patients with resected NSCLC at the study institution were investigated. Postoperative survival rates were estimated based on demographic and clinicopathologic factors, including Ki-67 expression and preoperative tobacco smoking habits. RESULTS: The overall postoperative 5-year survival rate in patients with high Ki-67 labeling indices (>/= 20%) was 39.6% compared with 67.7% in patients with low Ki-67 labeling indices. This finding was significant for all resected cases and for each pathologic disease stage (P < 0.05). The postoperative 5-year survival rate in patients with a history of heavy smoking (>/= 30 pack-years) was 47.6% compared with 62.5% for other patients (P = 0.027). This result was especially significant in patients with International Union Against Cancer Stage I disease and in patients with nonsquamous cell carcinoma (P < 0.03). The authors also observed a positive correlation between the Ki-67 labeling index and preoperative smoking habits (P = 0.0002). Multivariate analysis demonstrated that lymph node involvement, tumor differentiation, and Ki 67 labeling index were significant prognostic factors in NSCLC (P < 0.01). CONCLUSIONS: Tumor Ki-67 expression is a strong prognostic factor in NSCLC, especially adenocarcinoma. It may be hypothesized that tobacco mutagenicity may play a role in the growth and extension of NSCLC, which is one of the major impediments to postoperative survival in patients with a history of heavy smoking. PMID- 11013359 TI - Expression of connective tissue growth factor in cartilaginous tumors. AB - BACKGROUND: Connective tissue growth factor (CTGF) predominantly is expressed in hypertrophic chondrocytes and its specific receptors are demonstrated on chondrocytic cells. Therefore, CTGF may be involved in the proliferation and/or differentiation of cartilage cells. In the current study, CTGF expression was examined both in chondrosarcoma and enchondroma to clarify the relation between the expression of CTGF and the grade of malignancy. METHODS: The expression of CTGF and proliferating cell nuclear antigen (PCNA) were analyzed immunohistochemically in 34 cartilaginous tumor specimens. Eighteen tumors were determined to be chondrosarcoma including 8 Grade 1 tumors, 6 Grade 2 tumors, and 4 Grade 3 tumors. The percentage of CTGF positive and PCNA positive cells was quantified using at least 500 cells. RESULTS: CTGF was expressed in 70.1% of enchondroma cells, 84.0% of Grade 1 chondrosarcoma cells, 53.7% of Grade 2 tumor cells, and 26.8% of Grade 3 tumor cells (rho = -0.501; P = 0.0053). In chondrosarcoma cases, CTGF expression was correlated closely with tumor grade (rho = -0.920; P = 0.0001). There was a strong correlation between PCNA expression and tumor grade (rho = 0.907; P < 0.0001) and a strong negative correlation between CTGF and PCNA expression (rho = -0.493; P = 0.0061). In chondrosarcoma cases, patients with high expression of CTGF (>/= 30%) showed higher overall survival compared with those with low expression (< 30%) (P = 0.004). CONCLUSIONS: The current study revealed a correlation between the histologic grade of chondrosarcoma and prognosis, and the concomitant association between CTGF immunostaining and tumor grade and prognosis. Therefore, immunohistochemical staining with CTGF is a useful procedure for assessing the tumor grade and clinical course in patients with chondrosarcoma. PMID- 11013360 TI - Circulating CD44 and intercellular adhesion molecule-1 levels in low grade non hodgkin lymphoma and B-cell chronic lymphocytic leukemia patients during interferon-alpha-2a treatment. AB - BACKGROUND: Despite published reports regarding the association of elevated circulating CD44 and CD54 levels with unfavorable outcome in non-Hodgkin lymphoma (NHL) and chronic lymphocytic leukemia (CLL), little is known about therapy related changes in either adhesion molecule. In an attempt to evaluate the effect of interferon-alpha (IFNalpha), the authors measured serum CD44 (sCD44) and sCD54 in 22 low grade NHL and 14 CLL patients. METHODS: Twenty-six patients fulfilling the criteria for therapy received daily doses of 3 x10(6) IU IFNalpha-2a. Ten patients not requiring therapy also were observed for the same period. Fasting sera were collected at baseline and in 4 monthly intervals from all patients including the IFNalpha-treated (Group 1) and the untreated group (Group 2). RESULTS: Higher baseline sCD44 values were observed in Group 1 as compared with the Group 2 patients (P = 0.057). Within Group 1, patients were further divided between those who responded to IFNalpha, referred to as responders, and those who did not respond to IFNalpha, known as nonresponders. Responders showed a gradual decrease in sCD44 starting at the 4th month until the 12th month (P = 0.003, P = 0.002, and P = 0.01). Neither a difference in baseline nor an IFNalpha effect on the sCD54 levels could be shown. Soluble CD44 levels between responders and nonresponders were not different at the baseline but were significantly lower in responders, starting at month 4 and continuing throughout the therapy period (P = 0.04, P = 0.017, and P = 0.043). This decrease did not accompany a leukocyte or lactate dehydrogenase decrease and was not correlated with an early disappearance of the clinical findings. Similarly, after treatment, the sCD44 values of Group 1 were lower than the Group 2 levels at months 4 and 8 (P = 0.007 and P = 0.05, respectively). CONCLUSIONS: Soluble CD44, but not sCD54, can be used for monitoring therapy in patients with low grade NHL and CLL who have received IFNalpha, and for deciding whether further IFN therapy is necessary for those patients who have not responded to IFNalpha over a previous 12-month period. PMID- 11013361 TI - Avascular necrosis of the femoral head in chronic myeloid leukemia patients treated with interferon-alpha: a synergistic correlation? AB - BACKGROUND: The objectives of this study were to describe cases of avascular necrosis of the femoral head (ANFH) observed in chronic myeloid leukemia (CML) patients who were treated with interferon-alpha and to review the literature. METHODS: The authors undertook a case review of the M. D. Anderson experience with ANFH occurring in CML patients who were managed with interferon-alpha-based therapy. MEDLINE (from 1966 to November 1999) and CancerLit (from 1983 to November 1999) searches were conducted to identify cases of avascular necrosis (AVN) associated with either CML or interferon-alpha. RESULTS: Three patients with ANFH were identified from the authors' experience. No common features related to the disease or therapy were seen among them, except for the presence of thrombocytosis and loss of response. A literature review revealed seven cases of ANFH associated with CML with or without interferon-alpha-based therapy. ANFH was not reported in association with interferon-alpha use for indications other than the treatment of patients with CML. CONCLUSIONS: ANFH may be the result of an interaction between CML and interferon-alpha therapy. ANFH that occurs in patients with CML who are treated with interferon-alpha should be recognized for treatment implications. Thrombocytosis with consequent microvascular thrombi and avascular necrosis manifesting in susceptible vascular or weight-bearing areas (e.g., the femoral head) may be an associated finding along with loss of response to interferon-alpha therapy. PMID- 11013362 TI - Intermediate dose recombinant interferon-alpha as second-line treatment for patients with recurrent cutaneous melanoma who were pretreated with low dose interferon. AB - BACKGROUND: Interferon (IFN) is widely considered the most effective agent in the adjuvant therapy of patients with cutaneous melanoma (CM). However, little is known about the effect of IFN on pretreated CM patients who experience disease recurrence. The authors conducted a Phase II study to determine whether intermediate doses of IFN could be beneficial for these patients. METHODS: A series of 24 consecutive CM patients who had undergone surgery for local, in transit, or lymph node disease recurrence during adjuvant therapy with low dose IFN (IFNalpha-2b, 3 million units [MU] per day, three times per week) were enrolled for second-line therapy with intermediate dose IFN (IFNalpha-2b, 10 MU per day) for one year. RESULTS: IFN was discontinued in 7 patients (29.2%) because of toxicity. Several patients complained of impairment in their daily activities. Progression of disease was registered in 17 patients (70. 8%), with a median disease free survival of 5.5 months (95% confidence interval, 3.4-14.2). The median follow-up for the 7 patients who did not experience disease recurrence was 15 months (range, 13-22 months). CONCLUSIONS: An increased dose of IFN as second-line adjuvant treatment was poorly tolerated and produced negative clinical outcomes in patients with CM. However, these patients probably were unresponsive to IFN regardless of the dosage level. In fact, the first adjuvant IFN treatment was ineffective in all patients. Thus, the key factor in the treatment of CM seems to be patient responsiveness to IFN rather than the total dosage achieved. PMID- 11013363 TI - Long term results of a randomized study by the Swedish Melanoma Study Group on 2 cm versus 5-cm resection margins for patients with cutaneous melanoma with a tumor thickness of 0.8-2.0 mm. AB - BACKGROUND: Large, prospective, randomized trials with long term follow-up are required to obtain an unbiased evaluation of the significance of resection margins in patients with cutaneous melanoma. METHODS: The Swedish Melanoma Study Group performed a prospective, randomized, multicenter study of patients with primary melanoma located on trunk or extremities and with a tumor thickness > 0.8 mm and 0.8 mm thick and 5 cm, were found to have a high rate of distant failure; such patients merit consideration of a trial that examines the efficacy of systemic therapy. PMID- 11013365 TI - Fiberoptic ductoscopy for patients with nipple discharge. AB - BACKGROUND: Breast carcinoma and precancer are thought to start in the lining of the milk duct or lobule, yet until recently, we have not had direct access to this area other than by blindly removing tissue by core biopsy or fine-needle aspiration. Fiberoptic ductoscopy (FDS) is an emerging technique allowing direct visual access to the ductal system of the breast through nipple orifice exploration. METHODS: We applied ductoscopy to 259 women who had nipple discharge, and we analyzed the visual findings, the cytological washings, and the subsequent histopathology. RESULTS: In 92 (36%) of these women, fiberoptic ductoscopy was successful in detecting an intraductal papillary lesion. Of these observed lesions, 68 (74%) were single papilloma, 21 (23%) were multiple discrete papillomas, and 3 (3%) were diffuse intraductal thickening which corresponded to diffuse papillomatosis on histopathological analysis. The overall positive predictive value of FDS screening was 83%. Of the lesions observed, 29.8% were located in the main (segmental) duct, 43.9% lesions in the first branch, 17.5% lesions in the second branch, 7.9% in the third branch, and 0.9% in the fourth branch. These lesions had an overall average distance of 2.7 cm from the nipple orifice. Ductal washings performed at the time of ductoscopy were effective at obtaining representative exfoliated ductal cells which could be evaluated for the presence of clumps (> 50 cells), clumps with atypia or single ductal cells. The presence of clumps with positive FDS increased the positive predictive value to 86%. CONCLUSIONS: Fiberoptic ductoscopy currently offers a safe alternative to ductography in guiding subsequent breast surgery in the treatment of nipple discharge. PMID- 11013366 TI - Preservation of the saphenous vein during inguinal lymphadenectomy decreases morbidity in patients with carcinoma of the vulva. AB - BACKGROUND: Traditional inguinal lymphadenectomy includes the removal of a portion of the saphenous vein. The authors hypothesized that preserving the saphenous vein would decrease morbidity without affecting treatment outcome. METHODS: A retrospective review of 83 patients with carcinoma of the vulva who underwent inguinal lymphadenectomy between 1990-1998 was performed. Postoperative short term and long term complications were evaluated. RESULTS: A total of 139 inguinal dissections were performed in 83 patients. The saphenous vein was preserved in 62 patients and ligated in 77 patients. The clinical characteristics of the patients, the operating time, and the estimated blood loss were not significantly different between the two groups. The incidence rate of short term complications including fever, seroma, phlebitis, lymphocyst, and deep venous thrombosis also was similar. Cellulitis occurred in 39% of the patients who underwent vein ligation compared with 18% of the patients who underwent a vein sparing procedure (P = 0.006). Short term (< 6 months) lower extremity lymphedema occurred in 70% of the vein-ligated group compared with 32% of the vein-spared group (P < 0. 001). Chronic edema (>/= 2 years) was present in only 3% of the patients who underwent saphenous vein preservation compared with 32% of those who underwent vein ligation (P = 0.003). Chronic lymphedema in the vein-spared group was observed in only one patient who received postoperative radiation. Overall, individuals with preservation of the saphenous vein were less likely to develop complications (56% vs. 23%; P < 0.001). There was no difference in the rate of incidence of recurrent disease between the two groups. CONCLUSIONS: Preservation of the saphenous vein during inguinal lymphadenectomy reduces both the short term and long term postoperative complications without affecting treatment outcome. The saphenous vein should be preserved routinely in patients undergoing inguinal lymphadenectomy. PMID- 11013367 TI - High growth fraction at 9 grays of radiotherapy is associated with a good prognosis for patients with cervical squamous cell carcinoma. AB - BACKGROUND: MIB-1, a murine monoclonal antibody, recognizes the Ki-67 antigen of routinely processed paraffin sections after microwave treatment. Cycling cells are positive for MIB-1 in their nucleus, and quiescent cells are negative for MIB 1. The MIB-1 labeling index represents the growth fraction of the cell population. METHODS: A total of 150 cervical biopsy specimens were taken from 75 consecutive patients with cervical squamous cell carcinomas before radiotherapy (RT) and at 9 grays (Gy) of RT and were investigated to analyze the correlation between the MIB-1 labeling index or the deltaMIB-1 labeling index and patient prognosis or local disease control. The deltaMIB-1 index was calculated as the MIB-1 index at 9 Gy minus the MIB-1 index before RT. RESULTS: The mean MIB-1 index was 38% (range, 11-67%) before RT and 54% (range, 15-85%) at 9 Gy. The MIB 1 index at 9 Gy was related to prognosis, including overall survival (P = 0.025), disease free survival (P = 0. 024), and metastasis free survival (P = 0.045). Patients who had a deltaMIB-1 index > 20% showed a trend toward a higher disease free survival rate compared with patients who had a deltaMIB-1 index < 20% (P = 0.093). Neither the MIB-1 index nor the deltaMIB-1 index was associated with local disease control. CONCLUSIONS: The high growth fraction at 9 Gy is regarded as a predictive factor for a good prognosis in patients with cervical squamous cell carcinoma patients who undergo RT alone. PMID- 11013368 TI - A model for predicting surgical outcome in patients with advanced ovarian carcinoma using computed tomography. AB - BACKGROUND: A reliable model for predicting the outcome of primary cytoreductive surgery may be a useful tool in the clinical management of patients with advanced epithelial ovarian carcinoma. METHODS: Forty-one women with a preoperative computed tomographic (CT) scan of the abdomen and pelvis and a histologic diagnosis of Stage III or IV epithelial ovarian carcinoma following primary surgery performed by one of nine gynecologic oncologists were identified from tumor registry databases. All CT scans were analyzed retrospectively using a panel of 25 radiographic features without knowledge of the operative findings. Patient demographics, surgical findings and outcome, Gynecologic Oncology Group performance status, and pre-operative serum CA125 values were collected from patient medical records. Residual disease measuring < or = 1 cm in maximal diameter was considered an optimal surgical result. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for each radiographic feature and clinical characteristic. Based on statistical probability of each factor predicting cytoreductive outcome, 13 radiographic features, in addition to performance status, were selected for inclusion in the final model. Each parameter was assigned a numeric value based on the strength of statistical association, and a total Predictive Index score was tabulated for each patient. Receiver operating characteristic (ROC) curve analysis was used to assess the ability of the model to predict surgical outcome. Statistical significance was evaluated using the Fisher exact test. RESULTS: Twenty of 41 patients (48.8%) underwent optimal cytoreduction to /= 2 cm), bowel mesentery involvement (>/= 2 cm), suprarenal paraaortic lymph nodes (>/= 1 cm), omental extension (spleen, stomach, or lesser sac), and pelvic sidewall involvement and/or hydroureter were most strongly associated with surgical outcome. Using the Predictive Index scores, a receiver operating characteristic curve was generated with an area under the curve = 0. 969 +/- 0.023. In the final model, a Predictive Index score >/= 4 had the highest overall accuracy at 92.7% and identified patients undergoing suboptimal surgery with a sensitivity of 100% (21/21). The specificity, or ability to identify patients undergoing optimal surgery, was 85.0% (17/20). The PPV of a Predictive Index score >/= 4 was 87.5% (21/24), and the NPV was 100%. The ability of this model to correctly predict surgical outcome was statistically significant (P < 0.001). CONCLUSIONS: In this model, a Predictive Index score >/= 4 demonstrated high sensitivity, specificity, PPV, and NPV, and was highly accurate in identifying patients with advanced epithelial ovarian carcinoma unlikely to undergo optimal primary cytoreductive surgery. The Predictive Index model may have clinical utility in guiding the management of patients with ovarian carcinoma. PMID- 11013369 TI - Noninvasive, microinvasive, and invasive mucinous carcinomas of the ovary: a clinicopathologic analysis of 40 cases. AB - BACKGROUND: Whether ovarian mucinous tumors with epithelial stratification of more than three cell layers in the absence of stromal invasion (i.e., carcinomas diagnosed according to the Hart and Norris criteria) should be placed in the same category as mucinous tumors with stromal invasion (i.e., unquestionable carcinomas) remains controversial. Because individual mucinous tumors frequently contain benign, borderline, and malignant components, the adequacy of sampling has been emphasized. METHODS: We examined 21 mucinous carcinomas with no destructive stromal invasion (MCNI), 4 mucinous carcinomas with microinvasion (MCMI) of < 2 mm, and 15 mucinous carcinomas with invasion (MCI) of > or = 2 mm. Tumors were diagnosed as MCNI according to Hart and Norris criteria (12 tumors) or when severe nuclear atypia was present (9 tumors). Cases of MCNI were selected for review if a section had been taken for each 2 cm or less of the tumor's greatest diameter. The International Federation of Gynecology and Obstetrics stage and follow-up data of each case were examined, and differences among MCNI, MCMI, and MCI were analyzed. RESULTS: All 21 patients with MCNI and all 4 patients with MCMI had Stage I disease; there was no recurrence or death in these cases. In contrast, 7 of 15 patients with MCI had Stage II or III disease, and 8 patients died. CONCLUSION: MCNI clearly should be distinguished from MCI and be classified as noninvasive carcinomas after the absence of destructive stromal invasion has been confirmed by examining a sufficient number of sections. PMID- 11013370 TI - Paclitaxel, cisplatin, and epirubicin first-line chemotherapy in stage III and IV ovarian carcinoma: long-term results of a phase II study. AB - BACKGROUND: The combination of paclitaxel with cisplatin or carboplatin has become the preferred chemotherapy regimen in the treatment of epithelial ovarian carcinoma. Anthracyclines also have activity in this disease. We conducted a Phase II study by using the combination of paclitaxel, cisplatin, and epirubicin for the treatment of advanced ovarian carcinoma. METHODS: Forty consecutive patients with optimally (n = 7) or suboptimally (n = 33) debulked advanced ovarian carcinoma (International Federation of Gynecology and Obstetrics (FIGO) Stage III or IV) were treated with paclitaxel, 135 mg/m(2), as a 3-hour intravenous infusion, cisplatin 75 mg/m(2) intravenously (i.v.), and epirubicin 50 mg/m(2) i.v. every 3 weeks on an outpatient basis. Granulocyte-colony stimulating factor was administered at a dose of 5 microg/kg/day on Days 5-9. RESULTS: Among 28 patients with measurable disease, 24 (86%%) achieved an objective response including 19 complete and 5 partial responses. Among 18 patients who underwent reassessment laparotomy, pathologic complete response was confirmed in 9 patients. At a minimum follow-up of 40 months, the median overall survival had not been reached whereas the median time to progression for all patients was 18.7 months. The median remission duration for women with measurable disease who responded to treatment was 14 months. The treatment was well tolerated without toxic deaths; the most common toxicity was Grade 3/4 neutropenia that occurred in 30% of patients. Significant neuropathy (Grade 2 or higher) developed in only 8% of patients. CONCLUSIONS: The combination of paclitaxel, cisplatin, and epirubicin is a well tolerated outpatient regimen with significant activity in the treatment of advanced epithelial ovarian carcinoma. PMID- 11013371 TI - The clinical value of serum concentrations of cancer antigen 125 in patients with primary fallopian tube carcinoma: a multicenter study. AB - BACKGROUND: Primary fallopian tube carcinoma (PFTC) is a rare disease, and data on the serum concentration of tumor marker cancer antigen 125 (CA 125) in patients with this disease are sparse. The authors assessed the clinical value of the serum concentration of CA 125 as a prognostic and monitoring marker in patients with surgically treated PFTC. METHODS: In a multicenter study, the concentration of CA 125 was measured in 406 serum samples from 53 patients with PFTC. The results were correlated with clinical data. RESULTS: The pretreatment median serum CA 125 level was 183 U/mL (range, 6.5-5440.0 U/mL) in patients with PFTC. In a univariate Cox regression model, tumor stage and serum CA 125 level were associated significantly with shortened disease free survival (P = 0.006 and P < 0.001, respectively) and with overall survival (P = 0.03 and P = 0. 001, respectively). Lymph node involvement, tumor grade, and patient age were not associated with the length of survival. A multivariate Cox regression model showed that pretreatment the serum CA 125 level was a prognostic factor of disease free and overall survival, independent of tumor stage (P = 0.005 and P = 0.01, respectively). The pretreatment serum CA 125 level was correlated with tumor stage (P < 0.001) but not with lymph node involvement (P = 0.8), histologic grade (P = 0.3), or patient age (P = 0.2). The serum CA 125 level during chemotherapy was correlated significantly with Gynecologic Oncology Group response criteria to chemotherapy (P = 0. 001). During the follow-up of patients, serum CA 125 levels reached sensitivity, specificity, positive predictive value, and negative predictive value of 92%, 90%, 67%, and 98%, respectively, for differentiating between no evidence of disease and the presence of recurrent disease. In 90% of the patients, an increase of serum CA 125 level preceded the clinical or radiologic diagnosis of recurrent disease with a median lead time of 3 months (range, 0.5-7.0 months). CONCLUSIONS: This is the largest study to date with respect to serum CA 125 levels in patients with PFTC. The current data indicate that the pretreatment serum CA 125 level is an additional independent prognostic factor of disease free and overall survival in patients with PFTC. The serum CA 125 level adequately defines the response to chemotherapy and displays good sensitivity and specificity characteristics during the follow-up of patients with PFTC. PMID- 11013372 TI - Protracted survival after resection of metastatic uveal melanoma. AB - BACKGROUND: The objective of this study was to evaluate the usefulness of resection of metastatic uveal melanoma and to analyze the characteristics of patients who may benefit from surgical intervention. PATIENTS AND METHODS Twelve patients underwent surgical removal of metastasis between 1976 and 1998. Data regarding primary uveal melanoma, systemic metastasis, surgical procedures, and outcomes were reviewed retrospectively. RESULTS: There were seven patients with liver metastases, two with lung metastases, one with brain metastasis, and two patients with metastases in the liver and other organs. Median time to systemic metastasis was 8 years. Seven of 12 patients were asymptomatic when they were found to have metastasis. Ten patients underwent complete resection of metastasis. No significant surgical complications were experienced. Median recurrence free and overall survival periods after complete resection were 19 months (range, 6-78 months) and greater than 27 months (range, 11-86 months), respectively. Recurrence free and overall 5-year survival rates of those patients were 15.6% and 53.3%, respectively. Three of these patients had no further systemic recurrence. All patients whose time to systemic metastasis was within 5 years developed further systemic recurrence within 2 years after surgery. In contrast, in 8 patients whose time to systemic metastases was greater than 5 years, 4 patients either were recurrence free or developed second metastasis more than 4 years after surgery. CONCLUSIONS: Complete surgical removal of metastatic uveal melanoma provided unexpectedly long survival without significant morbidity for the selected patients. These results are encouraging and justify a trial in which patients eligible for resection are randomized between standard treatment and surgery. PMID- 11013374 TI - Schwannoma with angiosarcoma. Report of a case and comparison with other types of nerve tumors with angiosarcoma. AB - BACKGROUND: Schwannoma with angiosarcomatous change is a rare tumor, the clinical characteristics of which have not been analyzed. METHODS: A patient with schwannoma with angiosarcoma arising in the midneck and clinically mimicking a carotid body paraganglioma is described with a literature review of all previously reported cases and a comparison of their clinical features with those of schwannoma with conventional malignant transformation and cases of neurofibroma and malignant peripheral nerve sheath tumor (MPNST) with angiosarcoma. RESULTS: There are four reported cases, including the present case. Schwannoma with angiosarcoma affects older adults, mainly men. Three tumors arose from the vagus nerve in the neck. Three of the four angiosarcomas were epithelioid in type. Treatment in all cases was surgical resection followed by radiation and chemotherapy in one case and by radiation alone in another. One patient died with residual local angiosarcoma 5 months after the diagnosis. The remaining three patients were alive and disease free at 27 months, 43 months, and 90 months, with distant metastasis (after 15 months) reported only in the patient described in this case report. CONCLUSIONS: Schwannoma with angiosarcoma should be included in the differential diagnosis of presumed carotid body paragangliomas. Like angiosarcoma alone and schwannoma with conventional malignant transformation, but unlike cases of neurofibroma and MPNST with angiosarcoma, the patients are older adults, and there is a male prevalence. Schwannoma with angiosarcoma is capable of local spread with a fatal outcome and of distant metastasis, but follow-up strongly suggests that these patients have a better prognosis than patients with neurofibroma or MPNST with angiosarcoma. Recommended treatment is attempted complete surgical resection followed by radiation therapy and chemotherapy, if it can be tolerated by the patient. PMID- 11013373 TI - A clinicopathologic reappraisal of brain stem tumor classification. Identification of pilocystic astrocytoma and fibrillary astrocytoma as distinct entities. AB - BACKGROUND: Brain stem tumors in children have been classified pathologically as low grade or high grade gliomas and descriptively as diffuse gliomas, intrinsic gliomas, midbrain tumors, tectal gliomas, pencil gliomas, dorsal exophytic brain stem tumors, pontine gliomas, focal medullary tumors, cervicomedullary tumors, focal gliomas, or cystic gliomas. METHODS: To search for a simplified and prognostic clinicopathologic scheme for brain stem tumors, the authors reviewed a consecutive cohort of patients younger than age 21 years with tumors diagnosed from 1980 through 1997. Pathology specimens and neuroimaging were classified by masked review. Statistical and survival analysis along with Cox proportional hazards regression was performed. RESULTS: Seventy-six patients were identified, with initial diagnostic magnetic resonance imaging available for 51 and pathology specimens for 48 patients. Twenty cases were classified histologically as pilocytic astrocytoma (PA), 14 as fibrillary astrocytoma (FA), and 14 as other tumors or indeterminate pathology. For all tumors, characteristics significantly associated with a worse survival rate were: symptom duration less than 6 months before diagnosis (P = 0.004); abducens palsy at presentation (P < 0.0001); pontine location (P = 0.0002); and engulfment of the basilar artery (P = 0.006). Pilocytic astrocytoma was associated with location outside the ventral pons (P = 0.001) and dorsal exophytic growth (P = 0.013); Fibrillary astrocytoma was associated with symptoms less than 6 months (P = 0. 006), abducens palsy (P < 0.001), and engulfment of the basilar artery (P = 0.002). Pilocytic astrocytoma showed 5-year overall survival (OS) of 95% (standard error [SE], 5%) compared with FA 1-year OS of 23% (SE, 11%;P < 0.0001). CONCLUSIONS: Brain stem tumors can be succinctly and better biologically classified as diffusely infiltrative brain stem gliomas-generally FA located in the ventral pons that present with abducens palsy, often engulf the basilar artery, and carry a grim prognosis-and focal brain stem gliomas-frequently PA arising outside the ventral pons, often with dorsal exophytic growth, a long clinical prodrome, and outstanding prognosis for survival. Our findings emphasize the individuality of PA as a distinct clinicopathologic entity with an exceptional prognosis. PMID- 11013375 TI - Malignant lymphoma in southern Taiwan according to the revised European-American classification of lymphoid neoplasms. AB - BACKGROUND: The purpose of the current study was to determine the distribution and relative frequency of each subtype of malignant lymphoma in southern Taiwan according to the revised European-American classification of lymphoid neoplasms (REAL). METHODS: The pathology files of a regional hospital in southern Taiwan for 1989-1998 were searched for malignant lymphoma, lymphoproliferative disorder, and Hodgkin disease (HD). The results of light microscopy, immunohistochemistry, and in situ hybridization for Epstein-Barr virus-encoded RNA (EBER) were correlated with clinical findings, and all cases were classified according to REAL. RESULTS: A total of 205 cases were analyzed retrospectively. There were 197 cases (96.1%) of non-Hodgkin lymphoma (NHL) and 8 cases (3. 9%) of HD. Among the 197 NHL cases, 161 (81.7%) were of B-cell lineage and 36 (18.3%) were of T /natural killer cell lineage. Diffuse large B-cell lymphoma, extranodal marginal zone lymphoma, and follicular lymphoma were the most common B-cell subtypes and represented 47.2%, 19.3%, and 6.1%, respectively, of all NHL cases. Among the 36 cases of T-/natural killer cell lineage, unspecified peripheral T-cell lymphoma (8.6%), T-/natural killer cell lymphoma (angiocentric lymphoma) (4.1%), and anaplastic large cell lymphoma (3.6%) were the most common subtypes. Seven of eight T-/natural killer cell lymphoma cases were positive for EBER. The eight cases of HD were classified as lymphocyte-rich classic (two cases), nodular sclerosis (two cases), and mixed cellularity (four cases) subtypes. Three of these eight cases were positive for EBER. CONCLUSIONS: To the authors' knowledge this study is the first in Taiwan using the REAL classification and it again confirms the different geographic distribution of the various subtypes of malignant lymphoma. The frequency of T-/natural killer cell lineage NHL in Taiwan is higher than that in Western countries but not as high as reported previously. PMID- 11013376 TI - Breast and cervical carcinoma mortality among women in the Appalachian region of the U.S., 1976-1996. AB - BACKGROUND: Previous studies have shown high cervical carcinoma mortality and increasing breast carcinoma mortality in the Appalachian region of the U.S. (which includes parts of 12 states and all of West Virginia). In the current study the authors report trends in breast and cervical carcinoma death rates among women in Appalachia for 1976-1996. METHODS: Death rates were calculated from information provided on death certificates and reported to the National Center for Health Statistics for Appalachian women and for women living elsewhere in the U.S. ("other U.S. women"). Trends were examined with joinpoint regression techniques overall and by age and race. Average annual mortality rates were calculated by state for 1992-1996 for each state's Appalachian and non Appalachian areas. RESULTS: Overall breast carcinoma mortality was lower among Appalachian women than among other U.S. women throughout the study period; however, after rates decreased among both groups in the 1990s, the difference appears to have narrowed. No such decline was observed for women age >/= 70 years. Overall cervical carcinoma mortality was higher among Appalachian women than among other U.S. women but decreased during the study period to rates closer to those for other U.S. women. No significant decrease was observed among women age < 50 years. Overall, for both black and white women, breast carcinoma mortality was lower and cervical carcinoma mortality higher among women in Appalachia compared with their counterparts elsewhere in the U.S. For both breast and cervical carcinoma, the average annual death rates (1992-1996) varied by geographic areas within the Appalachian states, but most differences were not significant. CONCLUSIONS: Analysis of mortality trends in breast and cervical carcinoma may provide guidance for prevention and control activities to reduce premature mortality from these diseases. PMID- 11013377 TI - Therapeutic use of interferon-alpha for lymphomatoid papulosis. AB - BACKGROUND: Lymphomatoid papulosis is a primary cutaneous, CD30 positive lymphoproliferative disorder with the potential to transform into systemic, malignant lymphoma. Therapeutic strategies for patients with lymphomatoid papulosis have been designed to prevent transformation but have proved to be either inefficacious or limited by side effects. METHODS: The authors compared the clinical, histologic, and immunohistochemical features from a group of five patients receiving interferon-alpha (IFN-alpha) subcutaneously three times per week with the same features from a group of six patients receiving conventional therapy, including photochemotherapy, antibiotics, topical corticosteroids, or surgery, in an open trial. RESULTS: In the IFN-alpha group, four patients showed a complete remission, and one patient showed a partial remission within a time period of 6 weeks. Two patients developed disease recurrences after discontinuation of short term IFN-alpha therapy (5-7 months). Thereof, one patient went into stable remission after long term IFN-alpha therapy (17 months), and one patient remains in partial remission. In the control group, one patient went into spontaneous remission, two patients showed partial remission, of which one patient developed progressive disease at a later time point, whereas three patients have recurrent disease despite of treatment. CONCLUSIONS: The current results indicate that the treatment with IFN-alpha of patients with lymphomatoid papulosis alters the clinical course of the disease with fewer side effects than previous regimens; however, short term treatment does not induce stable remission. Therefore, prolonged treatment appears to be warranted for these patients. PMID- 11013378 TI - Prophylactic action of oral fluconazole against fungal infection in neutropenic patients. A meta-analysis of 16 randomized, controlled trials. AB - BACKGROUND: Fluconazole is used widely for fungal prophylaxis. Although studies with bone marrow transplantation (BMT) recipients clearly showed the usefulness of oral fluconazole, results of the studies in neutropenic patients other than BMT recipients have been inconsistent. Therefore, the authors performed a meta analysis to evaluate the efficacy of fluconazole prophylaxis during chemotherapy induced neutropenia. METHODS: The authors identified reports that were not restricted to those in English and not restricted to published trials through MEDLINE, CANCERLIT, or the data base of the Pfizer company. The authors included prospective, randomized studies comparing oral fluconazole with placebo, no treatment, or oral polyenes as prophylaxis for fungal infections in neutropenic patients. Two independent authors extracted data from 16 trials with 3734 patients enrolled. The outcome measures were the development of fungal-related death, systemic and superficial fungal infections, the use of empiric intravenous amphotericin-B, and infections or colonization with fluconazole-resistant fungi. The summarized odds ratios (ORs) were calculated using the Mantel-Haenszel method and the DerSimonian-Laird method. RESULTS: Prophylactic fluconazole was not effective in reducing fungal-related death or in reducing proven, systemic fungal infections in non-BMT patients (OR, 0.91; 95% confidence interval [CI], 0.30-2.82 and OR, 0.85; 95% CI, 0.47-1.55, respectively). However, fluconazole was very effective in reducing superficial fungal infections (OR, 0.44; 95% CI, 0.24 0.80), even when it was given in lower doses (50-200 mg per day). There was no increase in proven, systemic infection of fluconazole-resistant fungi, although colonization of those fungi increased. When the results were combined in studies in which the incidence of systemic fungal infections was > 15%, fluconazole was effective in reducing such infections (OR, 0.23; 95% CI, 0.15-0.36). CONCLUSIONS: The current analyses failed to find an effect of fluconazole on both fatal fungal infection and systemic fungal infection in non-BMT patients. Further studies on severely neutropenic patients are warranted because prophylactic fluconazole seemed to be effective when the incidence of systemic fungal infection was expected to be > 15%. PMID- 11013379 TI - Usefulness of positron emission tomographic visualization for examination of in vivo susceptibility to metastasis. AB - BACKGROUND: Immune surveillance may play a role in protecting against the establishment of metastasis. The authors previously observed that the injection of as few as 10(4) lung metastatic B16BL6 melanoma cells/0.2 mL resulted in no metastasis and in a reduced rate of cell accumulation in the lung, the target organ. In the current study, the authors examined the correlation between metastatic potential and tumor cell trafficking by using a liver-metastatic model. METHODS: The liver-metastatic potential of RAW117-H10 cells was examined by varying the number of cells injected into mice through the portal vein. To investigate the trafficking of the cells, the authors performed positron emission tomography (PET) analysis, because advances in this technology now enable the use of PET to investigate the real-time trafficking of as few as 10(4) cells/0.2 mL. Furthermore, to clarify the role of the immune defense system, metastatic potential and cell trafficking also were examined by using macrophage-depleted mice. RESULTS: When 10(6) or 10(5) RAW117-H10 cells/0.2 mL were injected into mice, both quantities of cells caused liver metastasis, cells accumulated in the liver at a similar rate, and there was an approximately 10-fold difference in the number of accumulated cells between the two doses. However, the injection of 10(4) cells/0.2 mL did not produce metastasis, and the accumulation rate in the liver was less than one-tenth of that after the injection of 10(5) cells/0.2 mL. The treatment of mice with 2-chloroadenosine for depleting macrophages prior to the injection of 10(4) cells/0.2 mL resulted in the suppression of the fast elimination of the cells from the liver. Corresponding to this change in PET images, the injection of 10(4) cells/0.2 mL into 2-chloroadenosine-pretreated mice resulted in metastasis. CONCLUSIONS: The current study suggests that immune surveillance suppresses accumulation of tumor cells to the target and suppresses metastasis, and this effect is obvious when small numbers of tumor cells are used for the challenge. Furthermore, the immune defense system plays a role in the early stage of the metastatic process. PMID- 11013380 TI - Assessing symptom distress in cancer patients: the M.D. Anderson Symptom Inventory. AB - BACKGROUND: The purpose of this project was to develop the M. D. Anderson Symptom Inventory (MDASI), a brief measure of the severity and impact of cancer-related symptoms. METHODS: A list of symptoms was generated from symptom inventories and by panels of clinicians. Twenty-six symptoms and 6 interference items were rated by a validation sample of 527 outpatients, a sample of 30 inpatients from the blood and bone marrow transplantation service, and a cross-validation sample of 113 outpatients. Clinical judgment and statistical techniques were used to reduce the number of symptoms. Reliability, validity, and sensitivity of the MDASI were examined. RESULTS: Cluster analysis, best subset analysis, and clinical judgment reduced the number of symptoms to a "core" list of 13 that accounted for 64% of the variance in symptom distress. Factor analysis demonstrated a similar pattern in both outpatient samples, and two symptom factors and the interference scale were reliable. Expected differences in symptom pattern and severity were found between patients with "good" versus "poor" performance status and between patients in active therapy and patients who were seen for follow-up. Patients rated fatigue-related symptoms as the most severe. Groups of patients classified by disease or treatment had severe symptoms that were not on the "core" list. CONCLUSIONS: The core items of the MDASI accounted for the majority of symptom distress reported by cancer patients in active treatment and those who were followed after treatment. The MDASI should prove useful for symptom surveys, clinical trials, and patient monitoring, and its format should allow Internet or telephone administration. PMID- 11013382 TI - Treatment of atrophoderma of Pasini and Pierini-associated hyperpigmentation with the Q-switched alexandrite laser: a clinical, histologic, and ultrastructural appraisal. AB - BACKGROUND AND OBJECTIVE: Atrophoderma of Pasini and Pierini (APP) is an uncommon cutaneous disorder, with no known effective treatment, manifested by hyperpigmented patches that appear to be depressed compared with surrounding skin. This study investigated the effectiveness of the Q-switched alexandrite laser on a patient with extensive APP, and evaluated histopathologic and ultrastructural changes. STUDY DESIGN/MATERIALS AND METHODS: A man with stable APP underwent Q-switched alexandrite laser treatment to a patch on the trunk. Biopsies were obtained from treated and untreated sites of involvement. Light and transmission electron microscopic evaluation was performed to investigate melanosome number, size, and volume, as well as melanin granule number and size. RESULTS: After three treatment sessions, the treated area showed marked clinical improvement. Electron microscopy showed a 19% reduction in melanin granule number and size and a 65% reduction in melanosome number, size, and volume in larger melanosomes in treated compared with untreated sites. CONCLUSION: Treatment of APP with the Q-switched alexandrite laser results in clinical improvement. Electron microscopic evaluation suggests that the mechanism may be a reduction in the number, size, and volume of larger melanosomes as well as a decrement in melanin granule number and size. PMID- 11013381 TI - Optic nerve sheath fenestration with a novel wavelength produced by the free electron laser (FEL). AB - BACKGROUND AND OBJECTIVE: To determine whether 6.45-microm free electron laser (FEL) energy can successfully perform optic nerve sheath fenestration and to compare the acute and chronic cellular responses with this surgery. STUDY DESIGN/MATERIALS AND METHODS: Optic nerve sheath fenestration was performed in rabbits by using either FEL energy (< 2.5 mJ, 10 Hz, 325-microm spot size) or a knife. The optic nerve integrity and glial response were evaluated histologically acutely or 1 month postoperatively. RESULTS: The FEL at low energy effectively cut the optic nerve sheaths with minimal reaction in the underlying nerve. With FEL or knife surgical techniques, a mild astrocytic hypertrophy only adjacent to the fenestration was observed acutely in the glial fibrillary acidic protein (GFAP) -immunoreacted sections. The chronic healing responses after either technique appeared similar with: (1) a thin fibrous scar at the fenestration site, (2) cells uniformly distributed (hematoxylin and eosin), and (3) up regulation of GFAP and S100beta in astrocytes adjacent to the fenestration site. CONCLUSION: The FEL at low energy performs an optic nerve sheath fenestration in a small space with ease. Both FEL and knife incisions cause a similar rapid glial response near the fenestration site that remains 1 month later. PMID- 11013383 TI - Effect of Nd:YAG radiation at millisecond pulse duration on dentine crater depth. AB - BACKGROUND AND OBJECTIVE: The effect of laser parameters on laser-dentine interaction has not been explored fully. This in vitro study investigated the effect on dentine crater depth of Nd:YAG laser radiation with varying repetition rates, total delivered energy, and dentine site either dyed or undyed. STUDY DESIGN/MATERIALS AND METHODS: One hundred forty-four caries-free third molars were sectioned transversely to provide 288 upper and lower cut surfaces. The upper surfaces were dyed (IR5). These upper and lower cut surfaces were exposed to an Nd:YAG laser with a 7 millisecond (msec) pulse duration, pulse repetition rates (RR) of 2. 5, 5.4, and 10.5 Hz, and four total energies (2.28, 2.64, 3.6, 4.2 joules). Dentine crater depth was measured by using a Reflex microscope. Results were statistically analysed with analysis of variance. RESULTS: Crater depth increased with increase in total delivered energy and in dyed and inner dentine sites (P<0.0001) but decreased with increasing repetition rate (P< 0.0001). CONCLUSIONS: Increasing energy, dyed, and inner dentine sites produced deeper craters. PMID- 11013384 TI - Optimal light dose for interstitial photodynamic therapy in treatment for malignant brain tumors. AB - BACKGROUND AND OBJECTIVE: The primary goal was to determine the maximal tolerable light dose that can be administered to patients undergoing multifiber interstitial photodynamic therapy (PDT) of malignant brain tumors at a fixed dose of photosensitizer. STUDY DESIGN/MATERIALS AND METHODS: Eighteen patients (12 glioblastomas, 5 anaplastic astrocytoma, and 1 malignant ependymoma) were included in this study. The total light dose delivered to the tumor was divided into three groups of six patients each: 1,500-3,700 J, 3,700-4,400 J, and 4,400 5,900 J. RESULTS: Five patients (all glioblastomas) demonstrated postoperative permanent neurologic deficits. None of the patients in 1,500-3,700 J, two patients in 3,700-4,400 J, and three patients in 4,400-5,900 J had neurologic deficits. Glioblastomas recurred more often than anaplastic astrocytomas. Increasing the light dose did not make a difference in local/regional control of glioblastomas. Patients with anaplastic astrocytomas survived (mean, 493 days) longer than patients with glioblastomas (mean, 116.5 days) after PDT. Four patients had prolonged survival (more than a year) after PDT. CONCLUSIONS: Increasing the total light dose delivered to the tumor increases the odds of having a permanent neurologic deficit but does not increase survival or time to tumor progression. There was no difference in local or marginal recurrence with increasing light dose. Recurrent anaplastic astrocytomas tend to do better than recurrent glioblastomas with PDT. PMID- 11013385 TI - Significantly increased lesion size by using the near-infrared photosensitizer 5,10,15,20-tetrakis (m-hydroxyphenyl)bacteriochlorin in interstitial photodynamic therapy of normal rat liver tissue. AB - BACKGROUND AND OBJECTIVE: Penetration of tissues by activating light ultimately limits the size of the lesions achievable in interstitial photodynamic therapy. Measurements of the wavelength-dependence of tissue optical properties suggest that substantial improvements may be possible, particularly in pigmented organs such as the liver, by using drugs absorbing at near infrared wavelengths. STUDY DESIGN/MATERIALS AND METHODS: In this study, the extent of light induced necrosis with the photosensitive agents Photofrin (activated at 632 nm), meta tetra(hydroxyphenyl)chlorin (mTHPC) (activated at 652 nm) and 5,10,15,20 tetrakis(m-hydroxyphenyl)bacteriochlorin (mTHPBC) (activated at 740 nm) are compared in normal rat liver. Interstitial irradiation of mTHPBC-sensitized liver tissue resulted in significantly larger necrotic areas than irradiation of Photofrin and mTHPC-sensitised livers. CONCLUSION: The results illustrate the advantage of near-infrared photosensitizer activation and point to a specific role for mTHPBC in the interstitial treatment of liver tumours. PMID- 11013386 TI - Time-resolved fluorescence of human aortic wall: use for improved identification of atherosclerotic lesions. AB - BACKGROUND AND OBJECTIVE: This study characterized aortic time-resolved fluorescence spectra for stratified levels of atherosclerosis and proposed interpretation of spectrotemporal variations in terms of histologic changes. STUDY DESIGN/MATERIALS AND METHODS: Fluorescence emission transients were measured at 370-510 nm (337 nm excitation) on 94 excised human aortic samples, ranging from normal to advanced fibrous atherosclerotic lesion. Global analysis yielded a three-exponential approximation of the time-resolved spectra from which average lifetime and decay-associated spectra were derived. RESULTS: Average lifetime at 390 nm gradually increased from 2.4+/-0.1 nsec (normal aorta) to 3.9+/-0.1 nsec (advanced lesion). Fluorescence intensity was markedly decreased above 430 nm in intermediate and advanced lesions. Spectral intensity associated with the intermediate decay increased at 470-490 nm for early and intermediate lipid-rich lesions. CONCLUSION: Time-resolved fluorescence spectra of aortic samples presented distinctive features for each atherosclerotic lesion type, which could serve as characteristic markers for optical analysis of the aortic wall. PMID- 11013387 TI - In vitro effects of low-level laser irradiation at 660 nm on peripheral blood lymphocytes. AB - BACKGROUND AND OBJECTIVE: The effects of low-level laser light irradiation are still highly contested, and the mechanisms of its action still unclear. This study was conducted to test the effects of low-level laser irradiation at 660 nm on human lymphocytes and to investigate the possible mechanisms by which these effects are produced. STUDY DESIGN/MATERIALS AND METHODS: Whole blood obtained by phlebotomy was irradiated at 660 nm by using energy fluences between 0 and 5.0 J/cm(2). The lymphocytes were isolated after irradiation of the whole blood. For the control experiment, the lymphocytes were first isolated and then irradiated at the same wavelength and energy fluence for comparison. The proliferation of lymphocytes and the formation of free radicals and lipid peroxides were monitored. Hemoglobin was also irradiated in a cell-free environment to test for the production of lipid peroxides. RESULTS: Lymphocyte proliferation was significantly higher (P<0.05) as expressed by a Stimulation Index in samples irradiated in the presence of whole blood compared with lymphocytes irradiated after isolation from whole blood. Free radical and lipid peroxide production also increased significantly when samples were irradiated in the presence of red blood cells. CONCLUSION: The present study supports the hypothesis that one mechanism for the photobiostimulation effect after irradiation at 660 nm is the reaction of light with hemoglobin, resulting in oxygen radical production. PMID- 11013388 TI - New technique for producing hybridoma by using laser radiation. AB - BACKGROUND AND OBJECTIVE: With conventional methods, cell fusion rate is extremely low, and fusion of two specific cells is not possible. We developed a new method for inducing cell fusion under the microscope by using a microprocessing device by laser. STUDY DESIGN/MATERIALS AND METHODS: Under a microscope, the target cells were irradiated with laser beams for trapping. Then, the trapped cells were transferred and placed in contact with the corresponding cells, which were also fixed by laser beam. The pulse laser beams are focused on the contact surface to cut small perforations for mutual communication of the cytoplasms. RESULTS: The fusion rate of mouse myeloma cells was 38%. The rate of hybridoma production of myeloma cell and lymphocyte was 2%. We confirmed the proliferation of the newly formed hybridoma in HAT medium and the production of immunoglobulin G. CONCLUSION: This new cell fusion method is characterized by production of hybridomas of target cells, lower cell toxicity, and a high rate of hybrid production. PMID- 11013389 TI - Optical-thermal simulation of human tonsillar tissue irradiation: clinical implications. AB - BACKGROUND AND OBJECTIVE: Mucosa intact laser tonsillar ablation is an alternative to conventional tonsillectomy. The efficacy of this procedure was demonstrated in canines, but establishing the safety of irradiating human tonsils is paramount. STUDY DESIGN/MATERIALS AND METHODS: An optical-thermal simulation of tonsillar tissue irradiation was previously developed, but the effect of varying parameters was not investigated. The tissue response to irradiation at 5 25 watts for 1 minute and 10 watts for 10 seconds to 162 seconds is simulated. RESULTS: At 15 watts and greater, the peak temperature is over 100 degrees C and the mucosal temperature is over 70 degrees C. At the depth of the tonsil, the temperature does not vary significantly. The peak temperature is at 1 mm. The radial temperature profile is not significantly altered by longer irradiation times. CONCLUSIONS: The optimal dosimetry parameters for irradiation of human tonsillar tissue at 805 nm with the MILTA technique is under 15 watts for approximately 1 minute. PMID- 11013390 TI - Aneuploidy, the somatic mutation that makes cancer a species of its own. AB - The many complex phenotypes of cancer have all been attributed to "somatic mutation." These phenotypes include anaplasia, autonomous growth, metastasis, abnormal cell morphology, DNA indices ranging from 0.5 to over 2, clonal origin but unstable and non-clonal karyotypes and phenotypes, abnormal centrosome numbers, immortality in vitro and in transplantation, spontaneous progression of malignancy, as well as the exceedingly slow kinetics from carcinogen to carcinogenesis of many months to decades. However, it has yet to be determined whether this mutation is aneuploidy, an abnormal number of chromosomes, or gene mutation. A century ago, Boveri proposed cancer is caused by aneuploidy, because it correlates with cancer and because it generates "pathological" phenotypes in sea urchins. But half a century later, when cancers were found to be non-clonal for aneuploidy, but clonal for somatic gene mutations, this hypothesis was abandoned. As a result aneuploidy is now generally viewed as a consequence, and mutated genes as a cause of cancer although, (1) many carcinogens do not mutate genes, (2) there is no functional proof that mutant genes cause cancer, and (3) mutation is fast but carcinogenesis is exceedingly slow. Intrigued by the enormous mutagenic potential of aneuploidy, we undertook biochemical and biological analyses of aneuploidy and gene mutation, which show that aneuploidy is probably the only mutation that can explain all aspects of carcinogenesis. On this basis we can now offer a coherent two-stage mechanism of carcinogenesis. In stage one, carcinogens cause aneuploidy, either by fragmenting chromosomes or by damaging the spindle apparatus. In stage two, ever new and eventually tumorigenic karyotypes evolve autocatalytically because aneuploidy destabilizes the karyotype, ie. causes genetic instability. Thus, cancer cells derive their unique and complex phenotypes from random chromosome number mutation, a process that is similar to regrouping assembly lines of a car factory and is analogous to speciation. The slow kinetics of carcinogenesis reflects the low probability of generating by random chromosome reassortments a karyotype that surpasses the viability of a normal cell, similar again to natural speciation. There is correlative and functional proof of principle: (1) solid cancers are aneuploid; (2) genotoxic and non-genotoxic carcinogens cause aneuploidy; (3) the biochemical phenotypes of cells are severely altered by aneuploidy affecting the dosage of thousands of genes, but are virtually un-altered by mutations of known hypothetical oncogenes and tumor suppressor genes; (4) aneuploidy immortalizes cells; (5) non-cancerous aneuploidy generates abnormal phenotypes in all species tested, e.g., Down syndrome; (6) the degrees of aneuploidies are proportional to the degrees of abnormalities in non-cancerous and cancerous cells; (7) polyploidy also varies biological phenotypes; (8) variation of the numbers of chromosomes is the basis of speciation. Thus, aneuploidy falls within the definition of speciation, and cancer is a species of its own. The aneuploidy hypothesis offers new prospects of cancer prevention and therapy. PMID- 11013391 TI - Stochastic simulation of processive and oscillatory sliding using a two-headed model for axonemal dynein. AB - Computer simulations have been used in an attempt to understand experimental observations of processive and oscillatory sliding by one or a few axonemal dyneins. A simple two-headed model has been examined using stochastic simulation methods. To explain the experimental observations, the model must be capable of taking backward steps, as well as forward steps, and there must be hysteresis in switching between forward or backward stepping. When the effects of Brownian movement on motor strain are included, it is not possible to obtain oscillations as regular as the experimental records by using motor strain to regulate switching between forward or backward stepping. PMID- 11013392 TI - Hypophosphorylated neurofilament subunits undergo axonal transport more rapidly than more extensively phosphorylated subunits in situ. AB - Axonal transport of neurofilaments (NFs) has long been considered to be regulated by phosphorylation. We present evidence that in optic axons of normal mice, the rate of NF axonal transport is inversely correlated with the NF phosphorylation state. In addition to 200 kDa NF-H and 145 kDa NF-M, axonal cytoskeletons from CNS contained a range of phospho-variants of NF-H migrating between 160-200 kDa, and of NF-M migrating at 97-145 kDa. While 160 kDa phospho-variants of NF-H have been well characterized, we confirmed the identity of the previously-described 97 kDa species as a hypophospho-variant of NF-M since (1) pulse-chase metabolic labeling confirmed the 97 kDa species to be a new synthesis product that was converted by phosphorylation over time into a form migrating at 145 kDa, (2) the 97 kDa protein reacted with multiple NF-M antibodies, including one specific for hypophosphorylated NF-M, and (3) dephosphorylation converted NF-M isoforms to 97 kDa. Autoradiographic analyses following metabolic radiolabeling demonstrated that hypophosphorylated NF-H and NF-M isoforms underwent substantially more rapid transport in situ than did extensively phosphorylated isoforms, while NF-H subunits bearing a developmentally delayed C-terminal phospho-epitope transported at a rate slower than that of total 200 kDa NF-H. Differential transport of phospho-variants also highlights that these variants are not homogeneously distributed among NFs, but are segregated to some extent among distinct, although probably overlapping, NF populations, indicating that axonal NFs are not homogeneous with respect to phosphorylation state. PMID- 11013393 TI - Preferential incorporation of tubulin into the junctional region of ciliary outer doublet microtubules: a model for treadmilling by lattice dislocation. AB - Even in the presence of colchicine or Taxol(R), sea urchin embryonic cilia undergo substantial steady-state turnover, with a rate of tubulin incorporation approaching half that seen in full regeneration [Stephens: Mol Biol Cell 8:2187 2198, 1997]. Preliminary experiments suggest that tubulin incorporates differentially into the most stable portion of the outer doublet, the junctional protofilaments [Stephens: Cell Struct Funct 24:413-418, 1999]. To explore this possibility further, embryos of the sea urchin Tripneustes gratilla, a ciliary length inducible system [Stephens: J Exp Zool 269:106-115, 1994a], were pulse labeled with (3)H leucine during steady-state turnover or induced elongation, followed by regeneration in the presence of unlabeled leucine. Cilia were isolated by hypertonic shock and fractionated into detergent-soluble membrane plus matrix, thermally-solubilized microtubule walls, and insoluble 9-fold symmetric remnants of A-B junctional protofilaments plus associated architectural elements. The fractions were resolved by SDS-PAGE and the specific activity of alpha-tubulin was determined. In cilia undergoing turnover or elongation during an isotope pulse, the specific activity of tubulin in the junctional region approximated that of precursor membrane plus matrix tubulin but surpassed that of the tubule wall by a factor of approximately 1.5. In cilia regenerated during an isotope chase, the specific activity of junctional tubulin exceeded that of both the membrane plus matrix and the tubule wall by a similar factor. These data indicate that tubulin is preferentially incorporated into junctional protofilaments during steady-state turnover, induced elongation and regeneration. A model for directional incorporation based on surface lattice discontinuities in the outer doublet is proposed. PMID- 11013394 TI - Centrosome-directed translocation of Weibel-Palade bodies is rapidly induced by thrombin, calyculin A, or cytochalasin B in human aortic endothelial cells. AB - To examine the possible role of the cytoskeleton in exocytosis of Weibel-Palade bodies (WPBs), we used double immunofluorescence and electron microscopy to study the spatial relationships between WPBs and main cytoskeletal elements in endothelial cells treated with secretagogue, such as thrombin, or cytoskeleton damaging agents. Unexpectedly, we have found that WPBs undergo rapid translocation towards the centrosome both in cells treated with thrombin and in those treated with cytochalasin B or calyculin A. Typically, 3 or 5 min after agent addition compact cluster of WPBs became visible near the microtubule organizing center (MTOC) in most endothelial cells in which a fivefold increase in WPBs localized in close proximity to the mother centriole had been detected. In both thrombin- and cytochalasin-treated cells that exhibit a noticeable depletion in WPBs compared to control cells, WPBs located at the cell periphery were found to colocalize with vimentin intermediate filaments, but not with microtubules. In contrast, there was precise colocalization observed between WPBs and microtubules in calyculin-treated cells in which all WPBs undergo centrosome directed translocation within 15 min after the agent addition. When vimentin filaments were induced to collapse to a perinuclear location by the microtubule disrupting agent demecolcine, WPBs also translocated to the perinuclear region, where numerous WPBs were found to be localized within the bundles of intermediate sized filaments. The data provide the first direct evidence that secretory granules utilize microtubule-based transport system to move in retrograde direction, i.e., away from the plasma membrane, towards the centrosome. We suggest that anterograde movement of WPBs is primarily dependent on their interaction with vimentin intermediate filaments. PMID- 11013395 TI - Association of SPARC (osteonectin, BM-40) with extracellular and intracellular components of the ciliated surface ectoderm of Xenopus embryos. AB - SPARC (Secreted Protein, Acidic, Rich in Cysteine) was detected by immunohistochemistry in the sensorial layer of the bilayered embryonic epidermis of Xenopus laevis during neurulation, when a subset of the sensorial cells are selected to differentiate into ciliated cell precursors. After the ciliated cells had intercalated into the outer layer and had undergone ciliogenesis, intense SPARC immunostaining was associated with the cilia and remained associated with the cilia throughout their persistence on the epidermis. Circumferential SPARC immunostaining was also detected at the interface between surface epithelial cells. Animal cap explants indicated that the embryonic activation of SPARC expression in the dorsal ectoderm does not require signaling from factors secreted by the underlying mesoderm. Immunoelectron microscopy revealed that SPARC is intimately associated with the 9 + 2 microtubule arrays of cilia. Our data indicate that SPARC plays a role in the development and function of the surface ciliated epidermis of Xenopus embryos. We propose that the counter adhesive activity of SPARC facilitates the intercalation of ciliary cell precursors to the surface epithelial layer, where its Ca(2+)-binding abilities promote cell-cell adhesion. Based on its association with ciliary microtubule arrays, we also propose that intracellular SPARC may play a role in regulating ciliary beat frequency and polarity. PMID- 11013396 TI - Linear extrapolation method of analyzing solvent denaturation curves. AB - The two most common methods of measuring the conformational stability of a protein are differential scanning calorimetry and an analysis of solvent denaturation curves by using the linear extrapolation method. In this article, we trace the history of the linear extrapolation method, review how the method is used to measure protein stability, and then discuss some of the other important uses. PMID- 11013397 TI - Large contributions of coupled protonation equilibria to the observed enthalpy and heat capacity changes for ssDNA binding to Escherichia coli SSB protein. AB - Many macromolecular interactions, including protein-nucleic acid interactions, are accompanied by a substantial negative heat capacity change, the molecular origins of which have generated substantial interest. We have shown previously that temperature-dependent unstacking of the bases within oligo(dA) upon binding to the Escherichia coli SSB tetramer dominates the binding enthalpy, DeltaH(obs), and accounts for as much as a half of the observed heat capacity change, DeltaC(p). However, there is still a substantial DeltaC(p) associated with SSB binding to ssDNA, such as oligo(dT), that does not undergo substantial base stacking. In an attempt to determine the origins of this heat capacity change, we have examined by isothermal titration calorimetry (ITC) the equilibrium binding of dT(pT)(34) to SSB over a broad pH range (pH 5. 0-10.0) at 0.02 M, 0.2 M NaCl and 1 M NaCl (25 degrees C), and as a function of temperature at pH 8.1. A net protonation of the SSB protein occurs upon dT(pT)(34) binding over this entire pH range, with contributions from at least three sets of protonation sites (pK(a1) = 5.9-6.6, pK(a2) = 8.2-8.4, and pK(a3) = 10.2-10.3) and these protonation equilibria contribute substantially to the observed DeltaH and DeltaC(p) for the SSB-dT(pT)(34) interaction. The contribution of this coupled protonation ( approximately -260 to -320 cal mol(-1) K(-1)) accounts for as much as half of the total DeltaC(p). The values of the "intrinsic" DeltaC(p,0) range from -210 +/- 33 cal mol(-1) degrees K(-1) to -237 +/- 36 cal mol(-1)K(-1), independent of [NaCl]. These results indicate that the coupling of a temperature-dependent protonation equilibria to a macromolecular interaction can result in a large negative DeltaC(p), and this finding needs to be considered in interpretations of the molecular origins of heat capacity changes associated with ligand-macromolecular interactions, as well as protein folding. PMID- 11013398 TI - Confirmation of a unique intra-dimer cooperativity in the human hemoglobin alpha(1)beta(1)half-oxygenated intermediate supports the symmetry rule model of allosteric regulation. AB - The contribution of the alpha(1)beta(1)half-oxygenated tetramer [alphabeta:alphaO(2)betaO(2)] (species 21) to human hemoglobin cooperativity was evaluated using cryogenic isoelectric focusing. The cooperative free energy of binding, reflecting O(2)-driven protein structure changes, was measured as (21)DeltaG(c) = 5.1 +/- 0. 3 kcal for the Zn/FeO(2) analog. For the Fe/FeCN analog, (21)DeltaG(c) was estimated as 4.0 kcal after correction for a CN ligand rearrangement artifact, demonstrating that ligand rearrangement does not invalidate previous conclusions regarding this species. In the context of the entire Hb cooperativity cascade, which includes eight intermediate species, the 21 tetramer is highly abundant relative to the other doubly-ligated species, providing strong support for the previously determined consensus partition function of O(2) binding and for the Symmetry Rule model of hemoglobin cooperativity (Ackers et al., Science 1992;255:54-63). Cooperativity of normal human hemoglobin is shown to depend on site-configuration, and not solely the number of O(2) bound, nor the occupancy of alpha vs. beta subunits. Verification of a unique contribution from the alpha(1)beta(1)doubly-oxygenated species to the equilibrium O(2) binding curve strongly reinforces the Symmetry Rule interpretation that the alpha(1)beta(1)dimer acts both as a structural and functional element in cooperative O(2) binding. PMID- 11013399 TI - Structural thermodynamics of a random coil protein in guanidine hydrochloride. AB - An important problem in protein folding is to understand the relationship between the structure of a denatured ensemble and its thermodynamics. Using 0 - 6M GdnHCl at fixed pH, we evaluated dimensional changes of an extensively denatured ensemble along with a thermodynamic parameter (Deltaupsilon) that monitors the proton inventory of the ensemble. Reduced and carboxyamidated ribonuclease A (RCAM) is a member of a class of disulfide-free RNase A molecules believed to be random coils (extensively denatured) in aqueous solution. Because GdnHCl interacts more favorably with the protein than water does, this denaturant is observed to increase the Stokes radius of the random coil, with the greatest Stokes radius change occurring in the 0 - 1.5M GdnHCl range. Measurement of the degree of protonation (proton inventory) of the ensemble as a function of GdnHCl at the fixed pH shows that the thermodynamic character of the ensemble also changes markedly in the 0 - 1.5M GdnHCl range, but with little or no change beyond 1.5M GdnHCl. To obtain denaturant-independent DeltaG degrees (N-D) values, the linear extrapolation method (LEM) requires the thermodynamic character of the native and denatured ensembles to be invariant in the transition zone. The results reported here indicate that proteins with a transition midpoint in the 0 1.5M GdnHCl range will not give denaturant-concentration independent DeltaG degrees (N-D) values. Such LEM-derived DeltaG degrees (N-D) quantities are a property of the protein and the denaturant, a condition that considerably limits their value in understanding structural energetics. PMID- 11013400 TI - Energetics of the specific binding interaction of the first three zinc fingers of the transcription factor TFIIIA with its cognate DNA sequence. AB - The energetics of the specific interaction of a protein fragment (zf1-3) containing the three N-terminal zinc fingers of the Xenopus laevis transcription factor TFIIIA with its cognate DNA sequence, contained in a 15 bp DNA duplex were studied using isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC) and fluorescence titration. The use of both ITC and DSC is necessary to provide values for the thermodynamic parameters that have been corrected for thermal fluctuations of the interacting molecules. In the temperature range from 13 degrees C to 45 degrees C (where all the binding reaction components are folded), formation of the complex is enthalpically driven with a negative heat capacity effect (DeltaC(p)). In this respect, the binding reaction of zf1-3 is similar to those of other proteins that bind in the major groove of DNA. It is dissimilar to the association reactions of proteins, however, that bind in the minor groove of DNA and that are driven by a dominating entropy factor. Comparison of the experimental values of DeltaH(ass) and DeltaC(p) with expected values of these parameters, calculated from the burial of polar and nonpolar molecular surfaces, indicates that the polar groups at the protein/DNA interface are not completely dehydrated upon formation of the complex. It also seems that the expected large positive entropy of dehydration upon forming the zfl-3/DNA complex ( approximately 1900 J * K(-1) * mol(-1)) cannot be balanced by the reduction in translational/rotational and configurational freedom of the protein to the level of the observed entropy of binding (38 J * K(-1) * mol(-1)). It is suggested that the additional negative entropy contribution comes from a damping of torsional motions in the DNA duplex. PMID- 11013401 TI - Structural stability of binding sites: consequences for binding affinity and allosteric effects. AB - During the course of biological function, proteins interact with other proteins, ligands, substrates, inhibitors, etc. These interactions occur at precisely defined locations within the protein but their effects are sometimes propagated to distal regions, triggering highly specific responses. These effects can be used as signals directed to activate or inhibit other sites, modulate interactions with other molecules, and/or establish inter-molecular communication networks. During the past decade, it has become evident that the energy of stabilization of the protein structure is not evenly distributed throughout the molecule and that, under native conditions, proteins lack global cooperativity and are characterized by the occurrence of multiple independent local unfolding events. From a biological point of view, it is important to assess if this uneven distribution reflects specific functional requirements. For example, are binding sites more likely to be found in well structured regions, unstable regions, or mixed regions? In this article, we have addressed these questions by performing a structure-based thermodynamic stability analysis of non-structurally homologous proteins for which high resolution structures of their complexes with specific ligands are available. The results of these studies indicate that for all 16 proteins considered, the binding sites have a dual character and are characterized by the presence of regions with very low structural stability and regions with high stability. In many cases the low stability regions are loops that become stable and cover a significant portion of low molecular weight ligands upon binding. For enzymes, catalytic residues are usually, but not always, located in regions with high structural stability. It is shown that this arrangement provides significant advantages for the optimization of binding affinity of small ligands. In allosteric enzymes, low stability regions in the regulatory site are shown to play a crucial role in the transmission of information to the catalytic site. PMID- 11013402 TI - Thermodynamic analysis of interactions between denaturants and protein surface exposed on unfolding: interpretation of urea and guanidinium chloride m-values and their correlation with changes in accessible surface area (ASA) using preferential interaction coefficients and the local-bulk domain model. AB - A denaturant m-value is the magnitude of the slope of a typically linear plot of the unfolding free energy change DeltaG degrees (obs) vs. molar concentration (C(3)) of denaturant. For a given protein, the guanidinium chloride (GuHCl) m value is approximately twice as large as the urea m-value. Myers et al. (Protein Sci 1995;4:2138-2148) found that experimental m-values for protein unfolding in both urea and GuHCl are proportional to DeltaASA(corr)(max), the calculated maximum amount of protein surface exposed to water in unfolding, corrected empirically for the effects of disulfide crosslinks: (urea m value/DeltaASA(corr)(max)) = 0.14+/-0.01 cal M(-1) A(-2) and (GuHCl m value/DeltaASA(corr)(max)) = 0.28+/-0.03 cal M(-1) A(-2). The observed linearity of plots of DeltaG degrees (obs) vs. C(3) indicates that the difference in preferential interaction coefficients DeltaGamma(3) characterizing the interactions of these solutes with denatured and native protein surface is approximately proportional to denaturant concentration. The proportionality of m values to DeltaASA(corr)(max) indicates that the corresponding DeltaGamma(3) are proportional to DeltaASA(corr)(max) at any specified solute concentration. Here we use the local-bulk domain model of solute partitioning in the protein solution (Courtenay et al., Biochemistry 2000;39:4455-4471) to obtain a novel quantitative interpretation of denaturant m-values. We deduce that the proportionality of m value to DeltaASA(corr)(max) results from the proportionality of B(1)(0) (the amount of water in the local domain surrounding the protein surface exposed upon unfolding) to DeltaASA(corr)(max). We show that both the approximate proportionality of DeltaGamma(3) to denaturant concentration and the residual dependence of DeltaGamma(3)/m(3) (where m(3) is molal concentration) on denaturant concentration are quantitatively predicted by the local-bulk domain model if the molal-scale solute partition coefficient K(P) and water-solute exchange stoichiometry S(1,3) are independent of solute concentration. We obtain K(P,urea) = 1.12+/-0.01 and K(P,GuHCl) = 1.16+/-0.02 (or K(P,GuH+) congruent with 1.48), values which will be useful to characterize the effect of accumulation of those solutes on all processes in which the water-accessible area of unfolded protein surface changes. We demonstrate that the local-bulk domain analysis of an m-value plot justifies the use of linear extrapolation to estimate ( less, similar 5% error) the stability of the native protein in the absence of denaturant (DeltaG(o)(o)), with respect to a particular unfolded state. Our surface area calculations indicate that published m-values/DeltaASA ratios for unfolding of alanine-based alpha-helical oligopeptides by urea and GuHCl exceed the corresponding m-value/DeltaASA ratios for protein unfolding by approximately fourfold. We propose that this difference originates from the approximately fourfold difference (48% vs. 13%) in the contribution of polar backbone residues to DeltaASA of unfolding, a novel finding which supports the long-standing but not universally accepted hypothesis that urea and guanidinium cation interact primarily with backbone amide groups. We propose that proteins which exhibit significant deviations from the average m-value/DeltaASA ratio will be found to exhibit significant deviations from the expected amount and/or average composition of the surface exposed on unfolding. PMID- 11013403 TI - Protein heat capacity reflects the dynamics of enthalpy exchange between the single macromolecule and the surroundings. AB - Heat capacity has played a prominent role in relating macroscopic and microscopic properties of small molecules and crystals. However, its diagnostic power can also be used for macromolecules such as proteins. It is shown in the present study that the macroscopically observed protein heat capacity provides direct access to the thermodynamic state of the single protein molecule. The new model of the physical basis of protein heat capacity emphasizes the dynamic nature of protein molecules. It incorporates equilibrium fluctuations as an integral constituent and shows that the increase in the magnitude of equilibrium fluctuations is coupled to an increase in the enthalpy flux between the individual protein molecule and its surroundings. PMID- 11013404 TI - Binding of small organic molecules to macromolecular targets: evaluation of conformational entropy changes. AB - The conformational entropy is the largest unfavorable effect that must be overcome during protein folding and binding. Accurate predictions of protein stability and binding affinity require a precise way of evaluating conformational entropy changes. Previously we implemented a computational approach aimed at estimating conformational entropy changes in peptides (D'Aquino et al., Proteins 1996;25:143-156; Lee et al., Proteins 1994;20:68-84). Here we extend this approach to estimate conformational entropy changes in molecules of pharmaceutical interest. Calculations were carried out for a set of 36 small organic molecules containing one dihedral angle and different functional groups around the central bond. Entropy changes were calculated for these molecules as the difference between the entropy of the free molecule and the entropy of the molecule when it is constrained to occupy a particular range of dihedrals, as in the bound state. Entropy changes for binding of larger molecules can be estimated assuming additivity on a per bond basis. Thus, the results presented here provide an initial toolbox of conformational entropy values in the form of a lookup table that can be used in the estimation of entropy changes associated with binding processes of more complex molecules. To facilitate their use, the values were parameterized in terms of the number and type of atoms neighboring each specific dihedral. Both methods, lookup table and parameterized equation, provide a very fast way of evaluating conformational entropy changes, making them suitable for fast screening algorithms. PMID- 11013405 TI - Induction of DNA adducts by several polychlorinated biphenyls. AB - It is known that lower-chlorinated biphenyls are metabolically activated to electrophilic quinoid species capable of binding to DNA. Also, certain metabolites are capable of redox cycling, thereby increasing oxidative stress in biological systems. In the present study, we tested mono-, di-, tri-, tetra-, penta-, hexa-, and heptachlorinated biphenyls for their ability to bind with DNA and to induce oxidative DNA damage. We present additional evidence that several PCB congeners form DNA adducts after metabolic activation, which can be detected by the nuclease P1- or butanol-enrichment procedures of the (32)P-postlabeling technique. Butanol and nuclease P1 enrichments showed different adduct recoveries, depending on the level of chlorination of the biphenyls. Application of the nuclease P1 enrichment showed that the incubation of 2-chloro-; 3, 4 dichloro-; 2,4,4'-trichloro-; 3,4,5-trichloro-; and 2,2',5, 5' tetrachlorobiphenyl with calf thymus DNA and liver microsomes from rats treated with phenobarbital, followed by oxidation with a peroxidase, produced five to eight different DNA adducts. For these lower-chlorinated biphenyls, butanol enrichment generally showed a lower recovery. For some higher substituted congeners (3,3',4,4', 5-pentachloro-, 2,2',3,4,4',5'-hexachloro-, 2,2',4,4',5, 5' hexachloro-, and 2,2',3,4,4',5,5'-heptachlorobiphenyl), after butanol enrichment a single dominant spot was observed, which was absent in the nuclease P1 procedure. After incubation of calf thymus DNA with either higher- or lower chlorinated PCB congeners, we were not able to detect significantly increased levels of oxidative DNA damage above background levels, measured as 8-oxo-7, 8 dihydro-2'deoxyguanosine. In view of the carcinogenicity of PCB mixtures in animals and the ability of PCB metabolites to bind covalently to DNA, rats were orally treated with a mixture of PCBs (Aroclor 1242). PCB-DNA adduct levels were analyzed in PCB target organs: liver, thymus, glandular stomach, spleen, testes, seminal vesicles and prostate DNA. In vivo PCB-DNA adducts could not be detected by either the butanol- or by the NP1-enrichment procedure in rat target tissue DNA. Also, no differences in oxidative DNA damage could be observed between PCB treated rats and controls. These results indicate a lack of DNA reactivity of PCB mixtures in vivo. PMID- 11013406 TI - Effect of a dCTP:dTTP pool imbalance on DNA replication fidelity in Friend murine erythroleukemia cells. AB - Nucleotide pool imbalances have been reported to affect the fidelity of DNA replication and repair in prokaryotic and eukaryotic cells. We have reported previously that the mutagen-hypersensitive thymidine kinase (TK)-deficient Friend erythroleukemia (FEL) cells (subclones 707BUF and 707BUE), have a more than sixfold increase in the dCTP:dTTP pool ratio when compared to that of wild-type, TK-positive (TK(+)) clone 707 cells. In this study we present the results of an investigation of the effect of the dCTP:dTTP pool imbalance on the accuracy of DNA replication within 707BUF cells. We examined the spontaneous mutation spectra occurring at the adenine phosphoribosyltransferase (aprt) locus within clone 707 (TK(+)) and 707BUF (TK(-)) FEL cells. Mutations recovered at the aprt locus in FEL cells comprised: base substitutions (43:73), frameshifts (14:13.5), and deletions (43:13.5) [clone 707 (TK(+)):707BUF (TK(-)), respectively, expressed as percentages]. A comparison of the mutation spectra obtained for the two cell lines did not reveal any significant increase in misincorporation of dCTP, the nucleotide in excess, in 707BUF (TK(-)) cells, during DNA replication synthesis. These data suggest that the dCTP:dTTP pool imbalance does not alter the fidelity of DNA replication synthesis in 707BUF (TK(-)) FEL cells. Rather, the predominance of GC --> AT transitions (53%) in the 707BUF (TK(-)) spectrum may reflect a reduced efficiency of repair by uracil DNA glycosylase of uracil residues within these cells. PMID- 11013407 TI - Cloning and expression of rat CYP2E1 in Saccharomyces cerevisiae: detection of genotoxicity of N-alkylformamides. AB - A cDNA coding for rat cytochrome P450 2E1 was cloned into the multicopy vector pYeDP60 and expressed in haploid RSY6 and diploid RS112 yeast strains of Saccharomyces cerevisiae under control of the GAL10-CYC1 promoter. Spectral and catalytic properties of the expressed 2E1 were examined in whole cells or microsomes of both strains. The level of CYP2E1 obtained in RS112 (200 pmol/mg microsomal protein) was the highest among CYP2E1 produced in the various expression systems. The monooxygenase activity in the microsomes of both strains, measured as aniline hydroxylase, was found comparable to that of control rat hepatic microsomes. In a reconstituted system in the presence of exogenous rat P450 reductase, their activity increased about 10-fold. When exposed to the carcinogen NDMA, a known 2E1 substrate, the recombination frequency determined in the 2E1-expressing RS112 cells was enhanced, in a dose-dependent manner, up to 20 fold. The exposure of the same cells to the hepatotoxic solvents, N-methyl- and N ethylformamide, resulted in an induction of recombination frequency, which was not observed in the void plasmid containing RS112 cells in the presence of S9 hepatic fractions from pyrazole-induced rats, as a specific exogenous metabolic activation system. These results demonstrate that the 2E1-expressing cells metabolize the two N-alkylformamides to genotoxic intermediates and, therefore, they provide an useful tool to study the bioactivation mechanism of potential P450 2E1 substrates. PMID- 11013408 TI - Telomere sequences at the novel joints of four independent amplifications in Saccharomyces cerevisiae. AB - Primary gene amplification, the mutation from one copy of a gene per genome to two or more genes per genome is a major mechanism of oncogene overexpression. We previously developed a system in the yeast Saccharomyces cerevisiae to phenotypically detect primary amplifications of a reporter cassette, ADH4:CUP1. We present here the sequence analysis of novel joints from four independent, spontaneous circular amplifications identified by the ADH4:CUP1 system. All four novel joints consist of C(1-3) A telomeric repeats joined to short (14- to 16-bp) CA-rich tracts between ADH4 and the telomere of chromosome VII. In three of the four amplifications, the telomeric sequence and the CA-rich tract that are joined in the amplification are normally located in inverted orientation to each other on chromosome VII. In the fourth amplification, the CA-rich tract on chromosome VII is joined to telomere sequences from another chromosome. We suggest that formation of these amplifications was initiated by recombination between these CA rich tracts and a telomere. The resulting dicentric chromosome could start a breakage-fusion-bridge cycle that could be resolved by the formation of a circular amplification structure. PMID- 11013409 TI - Formation of circular amplifications in Saccharomyces cerevisiae by a breakage fusion-bridge mechanism. AB - Primary gene amplification, the mutation from one gene copy per genome to two or more copies per genome, is a major mechanism of oncogene overexpression in human cancers. Analysis of the structures of amplifications can provide important evidence about the mechanism of amplification formation. We report here the analysis of the structures of four independent spontaneous circular amplifications of ADH4:CUP1 in the yeast Saccharomyces cerevisiae. The structures of all four amplifications are consistent with their formation by a breakage fusion-bridge (BFB) mechanism. All four of these amplifications include a centromere as predicted by the BFB model. All four of the amplifications have a novel joint located between the amplified DNA and the telomere, which results in a dicentric chromosome, and is adjacent to all the copies of the amplified DNA as predicted by the BFB model. In addition we demonstrated that two of the amplifications contain most of chromosome VII in an unrearranged form in a 1:1 ratio with the normal copy of chromosome VII, again consistent with the predictions of the BFB model. Finally, all four amplifications are circular, one stable endpoint for molecules after breakage- fusion-bridge. PMID- 11013410 TI - Use of genetically engineered Salmonella typhimurium OY1002/1A2 strain coexpressing human cytochrome P450 1A2 and NADPH-cytochrome P450 reductase and bacterial O-acetyltransferase in SOS/umu assay. AB - The major pathway of bioactivation of procarcinogenic heterocyclic aromatic amines (HCAs) is cytochrome P450 1A2 (CYP1A2)-catalyzed N-hydroxylation and subsequent esterification by O-acetyltransferase (O-AT). We have previously reported that an umu tester strain, Salmonella typhimurium OY1001/1A2, endogenously coexpressing human CYP1A2 and NADPH-P450 reductase (reductase), is able to detect the genotoxicity of some aromatic amines [Aryal et al., 1999, Mutat Res 442:113-120]. To further enhance the sensitivity of the strain toward HCAs, we developed S. typhimurium OY1002/1A2 by introducing pCW"/1A2:hNPR (a bicistronic construct coexpressing human P450 1A2 and the reductase) and pOA102 (constructed by subcloning the Salmonella O-AT gene in the pOA101-expressing umuC"lacZ gene) in S. typhimurium TA1535. In addition, as an O-AT-deficient strain, we developed the OY1003/1A2 strain by introducing pCW"/1A2:hNPR and pOA101 into O-AT-deficient S. typhimurium TA1535/1,8-DNP. Strains OY1001/1A2, OY1002/1A2, and OY1003/1A2 expressed, respectively, about 150, 120, and 140 nmol CYP1A2/l culture (in whole cells), and respective cytosolic preparations acetylated 15, 125, and > or = 0 nmol isoniazid/min/mg protein as the O-AT activities of cytosolic preparations, respectively. We compared the induction of umuC gene expression as a measure of genotoxicity and observed that the OY1002/1A2 strain was more sensitive than OY1001/1A2 strain toward the genotoxicity of 2-amino-1,4-dimethylimidazo[4,5-f]quinol ine(MeIQ), 2-amino-3 methylimidazo[4,5-f]quinoline (IQ),2-amino-3, 8-dimethylimidazo[4,5-f]quinoxaline (MeIQx),2-aminoanthracene, 2-amino-6-methyldipyrido[1,2-a::3,2'-d]i midazole,3 amino-1, 4-dimethyl-5H-pyrido[4,3-b]indole, and 3-amino-1-methyl-5H-pyrido[4, 3 a]indole. However, the genotoxicity of MeIQ, IQ, and MeIQx was not detected with the OY1003/1A2 strain. These results indicate that the newly developed strain OY1002/1A2 can be employed in detecting potential genotoxic aromatic amines requiring bioactivation by CYP1A2 and O-acetyltransferase. PMID- 11013411 TI - Immunoperoxidase detection of polycyclic aromatic hydrocarbon-DNA adducts in mouth floor and buccal mucosa cells of smokers and nonsmokers. AB - Tobacco smoking is a major risk factor for oral cancer; mouth floor and buccal mucosa are among the most and least cancer-prone subsites, respectively, in the oral cavity. We investigated the applicability of immunohistochemistry of smoking induced DNA adducts in oral cells for assessing the exposure to carcinogens, and estimating the risk for oral cancer. Polycyclic aromatic hydrocarbon (PAH)-DNA adducts were measured in mouth floor and buccal mucosa cells of smokers (n = 26) and nonsmokers (n = 22) by means of a semiquantitative immunoperoxidase assay. Smokers had elevated levels of PAH-DNA adducts compared to nonsmokers in their mouth floor cells (0.045 +/- 0.022 versus 0.022 +/- 0.016, P = 0.0008 arbitrary units of immunohistochemistry) as well as in their buccal mucosa cells (0.058 +/- 0.028 versus 0.028 +/- 0.012, P = 0.001). Also, there was a correlation between the levels of PAH-DNA adducts in mouth floor cells and those in buccal mucosa cells (r = 0.4, P = 0.01). Furthermore, PAH-DNA adduct levels in both mouth floor and buccal mucosa cells were significantly related to current smoking indices (amount of tar and number of cigarettes consumed per day). Expectedly, the levels of PAH-DNA adducts neither in mouth floor cells nor in buccal mucosa cells, both being short-lived cells, were related to smoking history index (pack years). The levels of PAH-DNA adducts, however, in mouth floor cells as the cancer prone cells were lower than those in buccal mucosa cells (0.037 +/- 0.023 versus 0.044 +/- 0.026, P = 0.04). We conclude that immunohistochemistry of PAH-DNA adducts in oral cells can be used for exposure assessment of tobacco-related carcinogens, however, it cannot be used for oral cancer risk estimation. PMID- 11013413 TI - Adaptation to alkylation damage in DNA measured by the comet assay. AB - The alkylating mutagens N-methyl-N-nitrosourea (MNU) and methyl methanesulfonate (MMS) were studied for their potential to induce DNA strand breaks and abasic (AP) sites in meristematic nuclei of Vicia faba root tips by the comet assay. The alkaline unwinding/neutral electrophoresis (A/N) and alkaline unwinding/alkaline electrophoresis (A/A) protocols were used for detection of DNA damage. With the A/N comet assay, less DNA damage was seen after conditioning pretreatment with a low dose prior to a high challenging dose of alkylating mutagens as compared to application of the high dose only, whereas a nearly additive effect was seen when the A/A comet assay was used. Adaptation was even more obvious when AP sites were revealed by the AP-endonuclease activity of exonuclease III. The adaptation observed with the A/N comet assay was abolished by pretreatment with the protein synthesis inhibitor cycloheximide. These data suggest that the comet assay is able to detect on molecular level a phenomenon resembling clastogenic adaptation. PMID- 11013412 TI - Frequency of minisatellite repeat number changes at the MS205 locus in human sperm before and after cancer chemotherapy. AB - To determine whether the measurement of repeat number mutations at a minisatellite locus could detect human germline mutations induced by chemotherapy, we performed a longitudinal study of the mutation frequencies in sperm from 10 patients treated for Hodgkin's disease. Polymerase chain reaction on small pools of DNA equivalent to 100 sperm and Southern blotting were used to screen at least 7900 sperm in each sample to quantify the mutation frequency at the minisatellite MS205 locus. Pretreatment and posttreatment semen samples were obtained at least 2 months after completion of therapy from 4 patients treated with a regimen (Novantrone, Oncovin, vinblastine and prednisone [NOVP]) that lacks alkylating agents and from three patients treated with regimens (Cytoxan, vinblastine, procarbazine and prednisone/Adriamycin, bleomycin, dacarbazine, lomustine, and prednisone [CVPP/ABDIC] or mechlorethamine, Oncovin, procarbazine and prednisone [MOPP]) containing alkylating agents. There were no effects of NOVP or CVPP/ABDIC on the mutation frequencies. In the 1 patient treated with MOPP, the treatment with the highest dose of gonadotoxic alkylating agents, there was a statistically significant increase in mutation frequency from 0.79% pretreatment to 1.14% posttreatment, indicating induction of mutations in stem spermatogonia. During-treatment semen samples obtained from 2 patients treated with ABVD, which does not contain gonadotoxic alkylating agents, and 1 with NOVP also did not show any increases above the baseline mutation frequencies, indicating no increase in the minisatellite mutation frequency in spermatocytes. Thus, measurement of repeat number changes at minisatellite MS205 appears to be able to detect induced germline mutations in human sperm. However, most chemotherapy regimens do not significantly increase this class of mutations. PMID- 11013414 TI - Absence of significant genotoxicity in lymphocytes and urine from workers exposed to moderate levels of cobalt-containing dust: a cross-sectional study. AB - Mortality studies have shown that, in the past, lung cancer occurred after exposure to mixtures of cobalt metal and metallic carbide particles, the main constituents of hard metals, but apparently not when exposure was to cobalt alone. The major objective of this biomonitoring study was to assess genotoxic effects as a measure for carcinogenic risk in workers from cobalt refineries and hard metal plants currently exposed to the threshold limit value/time-weighted average (TLV-TWA) for cobalt-containing dust. The study comprised three groups of workers: 35 workers exposed to cobalt dust from three refineries, 29 workers exposed to hard metal dust from two producing plants, and 35 matched control subjects recruited from the respective plants. The study design integrated complementary methodologies to assess biomarkers of effects that represent both initial DNA damage (8-hydroxydeoxyguanosine [8-OHdG] in urine and comet assay on lymphocytes) and definitive chromosome breakage/loss (micronuclei in lymphocytes). Cobalt and cotinine were determined in urine as a measure for cobalt exposure and recent smoking, respectively. No significant increase of genotoxic effects was detected in workers exposed to cobalt-containing dust as compared to controls. No difference in any genotoxicity biomarker was found between workers exposed to cobalt and hard metal dusts. Multiple regression analysis indicated that workers who smoked and were exposed to hard metal dusts had elevated 8-OHdG and micronuclei values. Because this observation is in line with a previous epidemiological study of an increased risk of dying from lung cancer in workers from the hard metal industry who smoked, it is concluded that this specific occupational group needs closer medical surveillance. PMID- 11013415 TI - Erratum PMID- 11013416 TI - Analysis of metal complex azo dyes by high-performance liquid chromatography/electrospray ionization mass spectrometry and multistage mass spectrometry AB - Five metal complex azo compounds were analyzed using negative-ion electrospray ionization mass spectrometry (ESI-MS). Mass spectra of all compounds yield intense peaks corresponding to [M - H](-) ions without any fragmentation, where M denotes the neutral compound with a proton as the counterion. Under collision induced dissociation (CID) conditions, structurally important fragment ions were studied using the ion trap analyzer with a multistage mass spectrometry (MS(n) facility. Synthesized compounds with (15)N atoms in the azo group facilitated the fragmentation pattern recognition. A reversed-phase high-performance liquid chromatography (HPLC) method using 5 mM ammonium acetate in 70% aqueous acetonitrile as mobile phase was developed making possible the separation of all complex compounds tested. The lower detection limits of the ESI-MS method are in the range 10-20 ng of each compound. The HPLC/ESI-MS method makes possible the monitoring of ligand exchange in aqueous solutions of metal complex azo dyes, and also investigation of the stabilities of the complexes in solution. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11013417 TI - Two-dimensional gel electrophoresis/matrix-assisted laser desorption/ionisation mass spectrometry of a milk powder. AB - Proteins in a commercial milk powder have been separated by two-dimensional gel electrophoresis and analysed by matrix-assisted laser desorption ionisation mass spectrometry. The mass spectrometric analyses were conducted in two steps: analysis of the intact proteins following their passive extraction into a suitable solvent mixture and analysis in reflectron mode of in situ digests of a number of gel spots. The combination of the two methods allowed a reliable identification of a number of proteins, including nine caseins as well as certain protein modifications including single/multiple phosphorylation, lactose-protein conjugates and Coomassie Brilliant Blue adducts. Analyses of the intact proteins prior to their in situ digestion contributed to a more efficient and reliable consultation of protein databases. PMID- 11013418 TI - Influence of water vapour on selected ion flow tube mass spectrometric analyses of trace gases in humid air and breath. AB - Selected ion flow tube mass spectrometry (SIFT-MS) detects and quantifies in real time the trace gases, M, in air/breath samples introduced directly into a flow tube. Inevitably, relatively large partial pressures of water vapour are introduced with the sample and the water molecules become involved in the ion chemistry on which this analytical technique depends. When H(3)O(+) ions are used as the precursors for chemical ionisation and SIFT mass spectrometric analyses of M, they generally result in the formation of MH(+) ions. Also, when water vapour is present the H(3)O(+) ions are partially converted to hydrated hydronium ions, H(3)O(+).(H(2)O)(1,2,3). The latter may act as precursor ions and produce new product ions like MH(+).(H(2)O)(1,2,3) via ligand switching and association reactions. This ion chemistry and the product ions that result from it must be accounted for in accurate analyses by SIFT-MS. In this paper we describe the results of a detailed SIFT study of the reactions involved in the quantification of acetone, ethyl acetate, diethyl ether, methanol, ethanol, ammonia and methyl cyanide by SIFT-MS in the presence of water vapour. This study was undertaken to provide the essential data that allows more accurate analyses of moist air and breath by SIFT-MS to be achieved. It is shown using our standard analysis procedure that the error of SIFT-MS quantification caused by the presence of water vapour is typically 15%. An improved analysis procedure is then presented that is shown to reduce this error to typically 2%. Additionally, some fundamental data have been obtained on the association reactions of protonated organic molecules, MH(+) ions, with water molecules forming MH(+).H(2)O monohydrate ions. For some types of M, reaction sequences occur that lead to the formation of dihydrate and trihydrate ions. PMID- 11013419 TI - Radial stratification of ions as a function of mass to charge ratio in collisional cooling radio frequency multipoles used as ion guides or ion traps. AB - Collisional cooling in radio frequency (RF) ion guides has been used in mass spectrometry as an intermediate step during the transport of ions from high pressure regions of an ion source into high vacuum regions of a mass analyzer. Such collisional cooling devices are also increasingly used as 'linear', two dimensional (2D) ion traps for ion storage and accumulation to achieve improved sensitivity and dynamic range. We have used the effective potential approach to study m/z dependent distribution of ions in the devices. Relationships obtained for the ideal 2D multipole demonstrate that after cooling the ion cloud forms concentric cylindrical layers, each of them composed of ions having the same m/z ratio; the higher the m/z, the larger is the radial position occupied by the ions. This behavior results from the fact that the effective RF focusing is stronger for ions of lower m/z, pushing these ions closer to the axis. Radial boundaries of the layers are more distinct for multiply charged ions, compared to singly charged ions having the same m/z and charge density. In the case of sufficiently high ion density and low ion kinetic energy, we show that each m/z layer is separated from its nearest neighbor by a radial gap of low ion density. The radial gaps of low ion population between the layers are formed due to the space charge repulsion. Conditions for establishing the m/z stratified structure include sufficiently high charge density and adequate collisional relaxation. These conditions are likely to occur in collisional RF multipoles operated as ion guides or 2D ion traps for external ion accumulation. When linear ion density increases, the maximum ion cloud radius also increases, and outer layers of high m/z ions approach the multipole rods and may be ejected. This 'overfilling' of the multipole capacity results in a strong discrimination against high m/z ions. A relationship is reported for the maximum linear ion density of a multipole that is not overfilled. PMID- 11013420 TI - Characterization of photodegradation products of alachlor in water by on-line solid-phase extraction liquid chromatography combined with tandem mass spectrometry and orthogonal-acceleration time-of-flight mass spectrometry. AB - On-line solid-phase extraction liquid chromatography in combination with mass spectrometry (MS), i.e. MS/MS and orthogonal-acceleration time-of-flight MS, was used for the characterization of photodegradation products of alachlor in river water. Various MS/MS scan functions were used, in particular the precursor-ion and the daughter-ion modes, to screen for degradation products with structures closely related to that of alachlor and to obtain information on characteristic fragments of the degradation products. Elemental compositions of compounds found and some of their fragments were calculated from the accurate mass information obtained with orthogonal-acceleration time-of-flight MS. Some ten degradation products could be characterized by combining various types of mass spectral information. Since quite a number of isomers were identified, structures of the degradation products were proposed by considering the most likely fragmentation patterns in MS/MS experiments. Degradation products of alachlor found in the current study were compared with those reported in the literature. PMID- 11013421 TI - Probing protein unfolding through monitoring cysteine alkylation by matrix assisted laser desorption/ionisation mass spectrometry. AB - Matrix-assisted laser desorption/ionisation mass spectrometry was used to monitor interaction between three proteins and two basic Immobiline chemicals (pK 10.3 and pK >12) commonly used in immobilised pH gradients (IPG). For two of the investigated proteins, the observed alkylation channels of the cysteine residues exhibited unmistakable response to their gradual denaturation following treatment with different concentrations (0-8 M) of two commonly used denaturants, urea and guanidine hydrochloride. Our assessment for protein unfolding is based on the number and relative intensity of the alkylation channels, yet the present mass spectrometry data are in good agreement with data based on optical rotatory dispersion, in which another approach was used to assess protein unfolding. Whether the present simple, fast and specific mass spectrometry method can be developed as a probe for monitoring folding/unfolding of cysteine-containing proteins can only be demonstrated by generating similar data for a larger number of proteins. PMID- 11013422 TI - Detection of new propofol metabolites in human urine using gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry techniques. AB - Using hyphenated analytical techniques, gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS), a study on minor propofol metabolites in human urine was conducted. These techniques allowed identification of two new phase I metabolites (2-(omega-propanol)-6 isopropylphenol and 2-(omega-propanol)-6-isopropyl-1,4-quinol). In addition, their four corresponding conjugates (three glucuronides and one sulphate) were detected. Thus in human urine at least eight conjugate metabolites are produced, derived from four different aglycones (propofol; 2, 6-diisopropyl-1,4-quinol; 2 (omega-propanol)-6-isopropylphenol and 2-(omega-propanol)-6-isopropyl-1,4 quinol). PMID- 11013423 TI - Fast, generic gradient high performance liquid chromatography coupled to fourier transform ion cyclotron resonance mass spectrometry for the accurate mass analysis of mixtures AB - Fast gradient high performance liquid chromatography (HPLC) has been combined with a commercially available Fourier transform ion cyclotron resonance (FTICR) mass spectrometer for the routine and high performance analysis of mixtures. With this combination we were able to separate and detect, under high mass accuracy conditions, a six-component drug mixture in less than 5 minutes. The fast gradients described are now possible due to the development of mechanically robust, ultra pure silica packing materials, which allow relatively high flow rates (ca. 1 mL/min for a 2 mm diameter column). For the six compounds present in the model mixture, relative mass errors of less than 1 ppm were obtained (based on an external calibration) providing sufficient mass accuracy to make unequivocal assignments of empirical formulae. Preliminary results of fast gradient HPLC/FTICR-MS/MS are also shown for the same six-component mixture. Copyright 2000 John Wiley & Sons, Ltd. PMID- 11013424 TI - High-throughput cytochrome P450 inhibition screening via cassette probe-dosing strategy. IV. Validation of a direct injection on-line guard cartridge extraction/tandem mass spectrometry method for simultaneous CYP3A4, 2D6 and 2E1 inhibition assessment. AB - A highly efficient direct injection on-line guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 3A4, 2D6 and 2E1 inhibition potential via cassette dosing of midazolam, dextromethorphan and chlorzoxazone using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for analysis by DI-GCE/MS/MS. Due to the novel use of an extremely short C(18) guard cartridge (4 mm in length), this method exhibits several advantages such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI GCE/MS/MS method demonstrates acceptable accuracy and precision for the simultaneous quantification of 1'-hydroxymidazolam, dextrorphan and 6 hydroxychlorzoxazone in microsomal incubations. The inhibition potential of CYP3A4, 2D6 and 2E1 has been evaluated using their known selective inhibitors. The IC(50) values measured by the cassette dosing approach (high-throughput) are consistent with those observed by an individual dosing regimen (conventional) and are all in good agreement with the literature values. The results suggest that the cassette probe-dosing strategy may provide an in vitro approach to minimize cost while maximizing throughput of CYP inhibition evaluation of new chemical entities in support of drug discovery and development. PMID- 11013425 TI - Fragmentation of 3,7-dialkyl-1,5-diphenyl-3,7-diazabicyclo[3.3. 1]nonan-9-ones under electron ionization. AB - A series of 3,7-dialkyl-1,5-diphenyl-3,7-diazabicyclo[3.3. 1]nonan-9-ones was prepared, and the details of their fragmentation under electron ionization (EI) were elucidated. The molecular ions of each compound under consideration were quite abundant in their EI spectra. Full-scan spectra exhibited a number of fragment ions which were clearly assigned using MS/MS and accurate mass measurements. The basic fragmentation of 3,7-dialkyl-1,5-diphenylbispidinones was due to the cleavage of C(1)-C(2) bond followed by a hydrogen migration similar to an odd-electron McLafferty rearrangement. Alternatively, the C(1)-C(2) bond cleavage was followed by the elimination of an imine molecule, Alk-N=CH(2). Further fragmentation resulted in ions at m/z 234 and 103, present in the spectra of all the compounds under study. The fragmentation pathways proposed in this paper are based on the substituent shifts, accurate mass measurements and collision-induced dissociation spectra of selected ions. The results of the present work can be useful in selecting the fragment ions suitable for identification and quantitation of bispidinones in biological matrices. PMID- 11013426 TI - Tandem mass spectrometry in the determination of 4-hydroxy-2-nonenal at the cellular level. PMID- 11013427 TI - Nuclear transfer and cell transplantation: making more with less. AB - Pig cloning can be achieved by transfer of nuclei of differentiated somatic cells into enucleated oocytes. Then, developing embryos are placed into surrogate mothers for further development to full term. Although cloned pigs offer the possibility of unlimited organ supply for compatible xenotransplantation in humans, the yield of a predictable number of offspring is still at an experimental phase. Spermatogonial stem cells from a fertile donor can be transplanted to the testes of infertile recipients and generate sperm. At present, results from xenogeneic spermatogenesis by transplantation indicate that porcine, bovine and equine spermatogonia find the interior of the seminiferous tubular environment not favorable for further differentiation into meiotic prophase spermatocytes. Spermatogenic cell transplantation is a promising experimental alternative for understanding the conditions required by both the donor cells and the recipient testis to coexist and even cooperate towards the full development of fertilizing sperm. PMID- 11013428 TI - Paternal exposure to cyclophosphamide dysregulates the gene activation program in rat preimplantation embryos. AB - Although there has been progress in determining the mechanisms by which maternal toxicant exposure affects progeny, there is little information on the actions of drugs administered to the father. We investigated the effects of pre-conceptional paternal exposure to cyclophosphamide, an anti-cancer agent, on embryonic gene activation in the rat. The male pronucleus was formed earlier in embryos sired by cyclophosphamide-treated male rats than in those sired by controls; early male pronucleus formation was followed by alterations in the gene activation program. BrUTP incorporation into RNA and Sp1 transcription factor immunostaining were increased and spread over both cytoplasmic and nuclear compartments in 2-cell embryos sired by cyclophosphamide-treated males compared to controls. Total RNA synthesis was constant in 1-8 cell embryos sired by drug-treated fathers, while in control embryos RNA synthesis increased four-fold to peak at the 4-cell stage. In 2-cell embryos sired by drug-treated males, the relative abundance of candidate imprinted genes was elevated significantly above control; a peak in the expression of these genes was not observed until the 8-cell stage in control embryos. Thus, paternal drug exposure temporally and spatially dysregulated rat zygotic gene activation, altering the developmental clock. PMID- 11013429 TI - Detection of PACH1, a nuclear factor implicated in the transcriptional regulation of meiotic and early haploid stages of spermatogenesis. AB - Spermatogenesis occurs in a series of well-defined stages and serves as an excellent model for lineage-specific cell development. Yet, little is known regarding the transcriptional mechanisms responsible for cell- and stage dependent gene regulation in the male germ line. The rat and mouse proenkephalin genes are expressed from an alternative, spermatogenic cell-specific promoter specifically in meiotically-active pachytene spermatocytes and early post-meiotic spermatids. This promoter thus serves as an excellent model for defining transcriptional regulators involved in germ line-specific gene expression in meiotic cells. Previous transgenic studies identified a proximal, 51 bp 5' flanking sequence containing two direct repeat elements that are absolutely required for in vivo proenkephalin promoter activity in spermatocytes and spermatids. Here, footprinting analyses were used to further delineate the specific interactions of a spermatogenic cell nuclear factor with the repeat elements within the proximal promoter region. This repeat-binding factor was also shown to be developmentally upregulated specifically in pachytene spermatocytes. Using Southwestern analysis, we have identified a unique nuclear protein enriched in pachytene spermatocytes that specifically recognizes the repeat elements within the proximal 5'-flanking sequence. We propose that this DNA binding factor, termed PACH1, is a key transcriptional regulator of the proenkephalin and potentially other gene promoters, uniquely expressed during meiosis in the male germ line. PMID- 11013431 TI - Organization of a gene coding for an oviduct-specific glycoprotein (oviductin) in the hamster. AB - Hamster oviductin, a high molecular weight glycoprotein secreted by the oviducts, is believed to participate in fertilization and protection of the tubal epithelium. Expression of the oviductin gene is confined strictly to nonciliated secretory cells of the oviduct and is regulated by hormones. The objective of this study was to characterize the genomic organization and to identify potential regulatory elements implicated in the control of transcription of the oviductin gene. Polymerase chain reaction was performed on hamster genomic DNA, yielding 2.2 kb of the 5' flanking region as well as 13.6 kb of genomic sequence comprising the entire coding sequence of the oviductin gene distributed in 11 exons. Sequencing of the 5' flanking region revealed, among other elements, an almost perfect estrogen-responsive element (GGTCACTGTGACT), an atypical TATA box (TATTAA), and a perfect inverted Sp1 site located between the transcription start site and the atypical TATA box. Primer extension analyses indicated that the hamster oviductin transcript possesses an unusually short 5' untranslated region of only 14 nucleotides. The distinct organization of the hamster oviductin gene in the vicinity of the transcription start site provides an interesting ground for further functional studies. PMID- 11013430 TI - Evaluation of heterologous insulator function with regard to chromosomal position effect in the mouse blastocyst and fetus. AB - Insulators are located at the boundaries of differentially regulated genes and delimit their interactions by establishing independent chromatin structures. Recently, an insulator sequence has been found in the 5'-flanking region of arylsulfatase (ARS) gene from sea urchin. To investigate functional conservation of this ARS insulator in mice, we performed blastocyst assays to evaluate the effect of this insulator on the chromosomal position effect, quantitatively. We constructed transgenes that have a luciferase gene under the control of the CMV IE enhancer and the human elongation factor 1 alpha promoter in the presence or absence of the ARS insulator in both flanking regions. These transgenes were microinjected into 1-cell mouse embryos and luciferase activity was measured at the blastocyst stage. We found that the presence of ARS insulator sequence doubled the number of luciferase-expressing blastocysts, and that the proportion of the blastocysts with high-level expression (> or = 1 x 10(4) relative light units (RLU)) was increased more than tenfold. In the case of transgenic fetuses, however, the presence of ARS insulator did not seem to improve transgene expression. These results suggest that the sea urchin ARS insulator confers position-independent expression driven by the human elongation factor 1 alpha promoter, at least in the blastocyst stage of the mouse. PMID- 11013432 TI - Preimplantation bovine embryos express mRNA of growth hormone receptor and respond to growth hormone addition during in vitro development. AB - In previous studies we demonstrated that bovine cumulus oocyte complexes (COCs) obtained from small and medium sized follicles express growth hormone receptor (GHR) mRNA and respond to growth hormone (GH) addition during in vitro maturation. The aim of this study was to investigate whether bovine zygotes and preimplantation embryos continue the expression of GHR gene after in vitro fertilization and during early embryo development and whether supplementation of GH during embryo culture affects embryo development. Therefore, COCs obtained from small and medium sized follicles were cultured in M199 supplemented with 10% FCS and gonadotropins for 24 hr. After in vitro fertilization the embryos were cultured: (a) on a monolayer of buffalo rat liver (BRL) cells in M199 supplemented with 10% FCS and 100 ng/ml bovine GH (NIH-GH-B18); (b) in droplets of serum-free BRL-conditioned medium supplemented with 100 ng/ml GH; (c) in droplets of synthetic oviductal fluid (SOF) supplemented with 100 ng/ml GH. Cultures without GH served as controls. Embryos were scored morphologically and the efficiency of the culture system was evaluated (a) as the percentage of cleaved embryos 4 days after IVF, (b) the percentage of blastocysts on Day 9 expressed on the basis of the number of oocytes at the onset of culture, and (c) the percentage of hatched blastocysts on Day 11 expressed on the basis of the total number of blastocysts present at Day 9. For gene expression, immature (GV) and mature (MII) oocytes (as positive control), embryos with less than 8 cells, 16-32 cells, and hatched blastocysts were prepared for reverse transcriptase polymerase chain reaction (RT-PCR) to assess the expression of mRNA of GHR. Messenger RNA for GHR was found in GV and MII oocytes and in all stages of embryo development. No mRNA for GH could be detected in early and expanded blastocysts produced in SOF medium. Immunoreactive GHR was found both in trophoblastic and embryonic cells of hatched blastocysts. Addition of 100 ng/ml GH during embryo culture on a monolayer of BRL cells in M199 supplemented with 10% FCS did not affect embryo development. However, GH (100 ng/ml) supplementation during embryo culture in droplets of serum-free BRL conditioned medium significantly (P < 0.05) enhanced the proportion of > 8-cell embryos. Similarly, culture of embryos in droplets of SOF medium in the presence of GH (100 ng/ml) significantly (P < 0.05) enhanced the number of > 8-cell embryos from 53.8% in control to 70.6% in GH treated group. Day 9 blastocyst formation in SOF medium was also significantly (P < 0.01) increased in the presence of GH (33.9%) compared to the control (20.2%). Embryos cultured in SOF without GH rarely resulted in hatched blastocysts (0.7%). However, GH supplementation remarkably enhanced the proportion of the hatched blastocysts (13%). In conclusion, expression of GHR gene in preimplantation bovine embryos, presence of the receptor, and the beneficial effect of GH on cleavage, blastocyst formation and hatchability of the embryos point to the involvement of GH in early embryonic development. PMID- 11013433 TI - EDTA stimulates cleavage stage bovine embryo development in culture but inhibits blastocyst development and differentiation. AB - Culture of bovine zygotes in medium SOFaa supplemented with 100 microM EDTA significantly increased cleavage rates during the first 72 hr of development compared to development in SOFaa. However, continued culture in the presence of EDTA for a further 72 hr (total of 6 days of culture) resulted in significantly reduced development to the morulae/blastocyst and blastocyst stages compared to culture without EDTA. Highest rates of development to the morulae/blastocyst stage (56.5%) and to the blastocyst stage (43.2%) were achieved when zygotes were cultured for 72 hr with EDTA before transfer to medium SOFaa without EDTA. Resultant blastocysts also had significantly increased blastocyst cell number and ICM cell number compared to those cultured without EDTA in the first 72 hr. EDTA was shown to inhibit glycolytic activity of the cleavage stage embryo, thereby preventing the premature stimulation of glycolysis and enhancing development. However, EDTA should not be used for the later stage embryo as the inhibition of glycolysis reduces energy production at the blastocyst stage and significantly inhibits inner cell mass development. PMID- 11013434 TI - In vitro development of porcine nuclear transfer embryos constructed using fetal fibroblasts. AB - The in vitro development of porcine nuclear transfer embryos constructed using primary cultures from day 25 fetal fibroblasts which were either rapidly dividing (cycling) or had their cell-cycle synchronized in G0/G1 using serum starvation (serum-starved) was examined. Oocyte-karyoplast complexes were fused and activated simultaneously and then cultured in vitro for seven days to assess development. Fusion rates were not different for either cell population. The proportion of reconstructed embryos that cleaved was higher in the cycling group compared to the serum-starved group (79 vs. 56% respectively; P < 0.05). Development to the 4-cell stage was not different using either population. Both treatments supported similar rates of development to the morula (1.5 vs. 7%, cycling vs. serum-starved) and blastocyst stage (1.5 vs. 3%, cycling vs. serum starved). The blastocyst produced using cycling cells had a total cell number of 10. Total cell numbers for the three blastocysts produced serum-starved cells were 22, 24, and 33. These blastocysts had inner cell mass numbers of 0, 15, and 4, respectively. Six hundred and thirty-five nuclear transfer embryos reconstructed using serum-starved cells were transferred to 15 temporarily mated recipients for 3-4 days. Of these, 486 were recovered (77% recovery rate) of which 106 (22%) had developed to the 4-cell stage or later. These were transferred to a total of 15 recipients which were either unmated or mated. Seven recipients farrowed a total of 51 piglets. Microsatellite analysis revealed that none of these were derived from the nuclear transfer embryos transferred. PMID- 11013435 TI - Germ cell transplantation from large domestic animals into mouse testes. AB - Donor-derived spermatogenesis after spermatogonial transplantation to recipient animals could serve as a novel approach to manipulate the male germ line in species where current methods of genetic modification are still inefficient. The objective of the present study was to investigate germ cell transplantation from boars, bulls, and stallions, which are economically important domestic animals, to mouse recipients. Donor testis cells (fresh, cryopreserved, or cultured for 1 month) were transplanted into testes of immunodeficient recipient mice in which endogenous spermatogenesis had been destroyed. Recipient testes were analyzed from 1 to > 12 months after transplantation for the presence of donor germ cells by donor-specific immunohistochemistry. Donor cells were present in most recipient testes with species-dependent differences in pattern and extent of colonization. Porcine donor germ cells formed chains and networks of round cells connected by intercellular bridges but later stages of donor-derived spermatogenesis were not observed. Transplanted bovine testis cells initially appeared similar but then developed predominantly into fibrous tissue within recipient seminiferous tubules. Few equine germ cells proliferated in mouse testes with no obvious difference between cells recovered from a scrotal or a cryptorchid donor testis. The pattern of colonization after transplantation of cultured cells did not resemble spermatogonial proliferation. These results indicate that fresh or cryopreserved germ cells from large animals can colonize the mouse testis but do not differentiate beyond the stage of spermatogonial expansion. Species-specific differences in the compatibility of large animal donors and mouse recipients were detected which cannot be predicted solely on the basis of phylogenetic distance between donor and recipient species. PMID- 11013436 TI - Maturational changes of the CD52-like epididymal glycoprotein on cynomolgus monkey sperm and their apparent reversal in capacitation conditions. AB - A major epididymal secretory protein in men has a colinear cDNA sequence with lymphocyte CD52, a sialylated glycoprotein. Immunostaining and flow cytometric detection of cynomolgus monkey sperm CD52 during epididymal maturation showed increases from 20 to 85% stained sperm from the caput to the corpus with staining intensities doubled. Freshly prepared cauda sperm showed only 10% staining while they markedly increased in percentage and intensity of staining upon incubation at 37 degrees C under capacitating conditions, but not at 4 degrees C. Western blotting of proteins from fresh cauda sperm revealed no less antigen than corpus sperm. Staining of ejaculated sperm exhibited similar increases during incubation. Further washing with a high salt medium before staining to remove any electrostatically-bound molecules masking the antigen showed no effect. Incubation-induced increases in antigen binding were accelerated by the addition of neuraminidase (0.25 and 0.5 U/ml), but not affected by the sialyl residue-rich fetuin (5 mg/ml) competing for any endogenous neuraminidase. There were no concomitant decreases in the staining of sialic acid residues during capacitation incubation. These findings suggest a cryptic antigen epitope site as a consequence of sperm maturation and subsequent re-exposure under capacitation conditions, but not due to the removal of sialic acid residues by endogenous neuraminidase. Involvement of endogenous proteases was also ruled out, as incubation in the presence of protease inhibitors did not hinder the increases but resulted in a dose-dependent enhancement in staining, suggesting some protease-sensitive unmasking process. In conclusion, the monkey epididymal secreted CD52 on sperm underwent changes in antigenic characteristics during sperm maturation which were reversed under capacitation conditions. PMID- 11013437 TI - Porcine oocyte activation induced by a cytosolic sperm factor. AB - It is not known how the fertilizing sperm elicits the release of Ca(2+) from the oocyte's intracellular stores. We investigated whether a crude extract isolated from boar sperm could induce the Ca(2+) release and trigger subsequent early and late activation events upon injection into matured porcine oocytes. The sperm extract induced an immediate rise in the intracellular free Ca(2+) concentration in all oocytes tested, which was followed by repetitive Ca(2+) transients in 11 out of 14 oocytes. Heat or trypsin treatment of the extract totally abolished the Ca(2+) releasing activity of the sperm factor. The injected oocytes showed cortical granule exocytosis, they resumed meiosis and entered first interphase: pronuclei were formed in 89.2% (132/148) of the cases. Pronuclear formation was accompanied by the appearance of a new 22 kDa protein as normally seen at fertilization. Of the successfully injected oocytes 51.7% (105/203) cleaved and 2.0% (4/203) developed to the blastocyst stage after being cultured for 7 days in NCSU 23 medium. Injection of the carrier medium could not trigger these changes. The results indicate that the sperm might activate porcine oocytes by introducing a soluble factor into the oocyte's cytoplasm after gamete fusion. PMID- 11013438 TI - Differential expression of glycoside residues in the mammalian zona pellucida. AB - The mammalian zona pellucida is an extracellular matrix surrounding the oocyte, and is composed of three major glycoproteins, ZP1, ZP2, and ZP3. Previous studies have suggested that the sperm receptor activity of the zona pellucida resides in specific oligosaccharide chains on the ZP3 glycoprotein. However, the nature of the terminal monosaccharide(s) on these glycosidic chains to which sperm bind is a matter of active debate. Evidence has been presented to support a role for at least three distinct monosaccharides in sperm binding, alpha-galactose, L-fucose on Lewis X structures, and beta-N-acetylglucosamine. Previous studies have shown that beta-N-acetylglucosamine is uniformly distributed throughout the zona matrix. In this study, we have investigated the expression and distribution of alpha-galactose and fucose moieties during the maturation of the zona pellucida in mouse, rat, and hamster. Interestingly, alpha-galactose residues are expressed only during later stages of zona secretion and, consequently, are confined to the inner portions of the mature zona pellucida in mouse and rat. In hamster, alpha galactose residues are only detectable in the zona pellucida of ovulated eggs, and are not found in ovarian oocytes. Fucosyl residues linked to Lewis X glycosides are not detectable at any stage of zona maturation in these three species, whereas fucose linked to N-linked core oligosaccharides are present throughout the zona. These studies indicate a previously unappreciated heterogeneity in the composition of zona glycosides. The specific localization of alpha-galactose residues to the inner portions of the zona matrix suggest a role in the later stages of sperm penetration through the zona. Finally, due to their absence from the zona surface, alpha-galactose and Lewis X fucosyl residues are not likely to be mediators of primary sperm binding. PMID- 11013439 TI - Mutations of the calcium-sensing receptor (CASR) in familial hypocalciuric hypercalcemia, neonatal severe hyperparathyroidism, and autosomal dominant hypocalcemia. AB - The calcium-sensing receptor (CASR) is a plasma membrane G protein coupled receptor that is expressed in the parathyroid hormone (PTH) producing chief cells of the parathyroid gland and the cells lining the kidney tubule. By virtue of its ability to sense small changes in circulating calcium concentration ([Ca(2+)](o)) and to couple this information to intracellular signaling pathways that modify PTH secretion or renal cation handling, the CASR plays an essential role in maintaining mineral ion homeostasis. Inherited abnormalities of the CASR gene located on chromosome 3p13.3-21 can cause either hypercalcemia or hypocalcemia depending upon whether they are inactivating or activating, respectively. Heterozygous loss-of-function mutations give rise to familial (benign) hypocalciuric hypercalcemia (FHH) in which the lifelong hypercalcemia is asymptomatic. The homozygous condition manifests itself as neonatal severe hyperparathyroidism (NSHPT), a rare disorder characterized by extreme hypercalcemia and the bony changes of hyperparathyroidism which occur in infancy. The disorder autosomal dominant hypocalcemia (ADH) is due to gain-of-function mutations in the CASR gene. ADH may be asymptomatic or present with neonatal or childhood seizures. A common polymorphism in the intracellular tail of the CASR, Ala to Ser at position 986, has a modest effect on the serum calcium concentration in healthy individuals. PMID- 11013440 TI - Genetic lesions of bilirubin uridine-diphosphoglucuronate glucuronosyltransferase (UGT1A1) causing Crigler-Najjar and Gilbert syndromes: correlation of genotype to phenotype. AB - Uridine-diphosphoglucuronate glucuronosyltransferases (UGTs) are a family of enzymes that conjugate various endogenous and exogenous compounds with glucuronic acid and facilitate their excretion in the bile. Bilirubin-UGT(1) (UGT1A1) is the only isoform that significantly contributes to the conjugation of bilirubin. Lesions in the gene encoding bilirubin-UGT(1), lead to complete or partial inactivation of the enzyme causing the rare autosomal recessively inherited conditions, Crigler-Najjar syndrome type-1 (CN-1) and type 2 (CN-2), respectively. Inactivation of the enzyme leads to accumulation of unconjugated bilirubin in the serum. Severe hyperbilirubinemia seen in CN-1 can cause bilirubin encephalopathy (kernicterus). Kernicterus can be fatal or may leave behind permanent neurological sequelae. Here, we have compiled more than 50 genetic lesions of UGT1A1 that cause CN-1 (including 9 novel mutations) or CN-2 (including 3 novel mutations) and have presented a correlation of structure to function of UGT1A1. In contrast to Crigler-Najjar syndromes, Gilbert syndrome is a common inherited condition characterized by mild hyperbilirubinemia. An insertional mutation of the TATAA element upstream to UGT1A1 results in a reduced level of expression of the gene. Homozygosity for the variant promoter is required for Gilbert syndrome, but not sufficient for manifestation of hyperbilirubinemia, which is partly dependent on the rate of bilirubin production. Several structural mutations of UGT1A1, for example, a G71R substitution, have been reported to cause mild reduction of UGT activity toward bilirubin, resulting in mild hyperbilirubinemia, consistent with Gilbert syndrome. When the normal allele of a heterozygote carrier for a Crigler-Najjar type structural mutation contains a Gilbert type promoter, intermediate levels of hyperbilirubinemia, consistent with the diagnosis of CN-2, may be observed. PMID- 11013441 TI - Characterization of two unusual RS1 gene deletions segregating in Danish retinoschisis families. AB - Over 100 distinct retinoschisis gene (RS1) mutations, of which approximately 10% are single exon deletions, have been described to date. In this paper we have characterized in detail two dissimilar RS1 gene deletions which are accountable for RS in one-third of Danish patients. First, a 136 kb deletion, spanning from the 5' region of the RS1 gene to intron 3, was identified. Unexpectedly this large deletion abolishes exons of three adjacent genes: serine-threonine phosphatase gene (PPEF-1)/serine-threonine protein phosphatase gene (PP7), retinoschisis gene (RS1), and serine-threonine kinase gene (STK9). We demonstrate that the RS1 and STK9 genes are partly overlapping and the sequences of the PP7 and PPEF-1 genes are identical. This is the first study which reports of retinoschisis patients who also suffer from deletions in genes adjacent to RS1. The 136 kb deletion is also the first gross deletion of the retinoschisis gene deleting three exons. It results from a recombination between two repetitive sequences of the Alu family, one in 5' region of the RS1 gene and the other in RS1 intron 3. The second alteration, the actual Danish RS founder mutation, is a 4.4 kb noncontiguous two-part deletion composed of two deleted 1.5 and 2.9 kb segments, separated by an intact 1.2 kb segment. It extends from the 5' flanking region of the retinoschisis gene to RS intron 1. RS1 gene deletions of this type have not been identified previously. Despite these two unique deletions, which either lead to severely defective transcription or total absence of the retinoschisin and PPEF-1 protein, all the patients have a typical retinoschisis phenotype. PMID- 11013442 TI - High mutation detection rate in TCOF1 among Treacher Collins syndrome patients reveals clustering of mutations and 16 novel pathogenic changes. AB - Twenty-eight families with a clinical diagnosis of Treacher Collins syndrome were screened for mutations in the 25 coding exons of TCOF1 and their adjacent splice junctions through SSCP and direct sequencing. Pathogenic mutations were detected in 26 patients, yielding the highest detection rate reported so far for this disease (93%) and bringing the number of known disease-causing mutations from 35 to 51. This is the first report to describe clustering of pathogenic mutations. Thirteen novel polymorphic alterations were characterized, confirming previous reports that TCOF1 has an unusually high rate of single-nucleotide polymorphisms (SNPs) within its coding region. We suggest a possible different mechanism leading to TCS or genetic heterogeneity for this condition, as we identified two families with no apparent pathogenic mutation in the gene. Furthermore, our data confirm the absence of genotype-phenotype correlation and reinforce that the apparent anticipation often observed in TCS families is due to ascertainment bias. PMID- 11013443 TI - Diverse deletions in the growth hormone receptor gene cause growth hormone insensitivity syndrome. AB - Growth hormone insensitivity syndrome (GHIS; also known as Laron syndrome), is characterized by severe postnatal growth failure and normal growth hormone. The syndrome is frequently caused by point mutations in the growth hormone receptor gene (GHR). Here we report five families with GHIS and partial deletions of the GHR gene. The deletion breakpoints were sequenced and PCR-based diagnostic tests were developed. In a Cambodian family, a novel deletion removed part of exon 5 and 1.2 kb of the preceding intron. The deletion occurred by recombination within four identical nucleotides. In the mutant transcript, skipping of the truncated exon 5 leads to a frameshift and premature termination codon (PTC). A previously reported discontinuous deletion of GHR exons 3, 5, and 6 was identified in three Oriental Jewish families. An unaffected individual was heterozygous for the exon 5 and 6 deletion, but homozygously deleted for exon 3 suggesting that the exon 3 deletion is a polymorphism. The pathogenic deletion of exons 5 and 6 spans about 7.5 kb. Sequence analysis of the breakpoints revealed an imperfect junction between introns 4 and 6, with a four basepair insertion. A novel deletion of 13 nucleotides within exon 9 was identified in a Caucasian girl with GHIS who carries the I153T missense mutation on her other allele. The exon 9 deletion leads to a frameshift and PTC. The predicted protein retains the transmembrane domain and a short cytoplasmic tail. Four family members in three generations were carriers of this deletion, but only two of them were below normal for height, suggesting that this mutation by itself does not act as a dominant negative, as was reported for two other GHR mutations which lead to truncation of the intracellular domain. PMID- 11013444 TI - Direct genomic multiplex PCR for BRCA1 and application to mutation detection by single-strand conformation and heteroduplex analysis. AB - Most mutation detection methods are based on analysis of PCR amplified segments and the application of multiplex PCR is one central approach to improving screening efficiency. Genes like the breast-ovarian cancer susceptibility gene BRCA1 pose a difficult challenge to efficient mutation screening because of large coding regions, numerous exons, and complex mutational spectra. The application to BRCA1 of a general approach to effective multiplex PCR is described here. Fifteen triplex PCRs and a single PCR reaction condition were used for amplification of all BRCA1 coding regions and the BRCA1-specific segments from the duplicated promoter region. SSCP/HDX gel analysis of the multiplex products detected mobility distinctions for 34/34 sets of allelic BRCA1 fragments. A novel polymorphism was found, CTTCT(4)CT(10)CT(12) >CT(4)CT(11), a compound deletion in a region beginning at the +33 position of IVS7 and resulting in a net deletion of 15 bp. This change was shown to be one of the common polymorphisms that define the two major haplotypes of the BRCA1-RNU2 region in a large proportion of the world population. A triplex PCR for SSCP detection of this deletion and two other distantly located common polymorphisms may be used to screen haplotype content and facilitate comparison of samples with similar haplotypes in subsequent mutation screening. The approach for robust multiplex amplification is generally applicable and allows rapid development of efficient testing for a wide variety of mutations in any gene(s) encompassing a large coding region or numerous exons and including as many as 50 different genomic PCR products. PMID- 11013445 TI - Identification of specific BRCA1 and BRCA2 variants by DHPLC. AB - Denaturing high performance liquid chromatography (DHPLC) is generating increasing interest in clinical genetics as a reliable tool for the analysis of genetic alterations. In the work presented here our intentions were to optimize primer design and DHPLC analysis conditions for a qualitative detection of BRCA1 and BRCA2 variations. The BRCA1 and BRAC2 genes display a high proportion of polymorphisms. Sequencing efforts geared towards the distinction of tumor-related mutations and benign variants still remain time-consuming and expensive. DHPLC elution profiles, however, permit the correlation of a characteristic chromatographic profile with a specific sequence alteration. In this study we evaluate the sensitivity of DHPLC for the identification of unique polymorphisms, which are frequent in the Caucasian population, in lieu of sequence analysis. The complete BRCA1 gene and parts of BRCA2 were examined. In the case of BRCA1, 431 out of 432 heterozygotes were identified correctly. In addition, 18 new profiles were identified which had not been detected previously in our studies and which represented new mutations or rare polymorphisms. For BRCA2, 135 out of 137 simple sequence variants were classified correctly. In addition, six new profiles were identified which represented new mutations or rare polymorphisms. PMID- 11013446 TI - Oligonucleotide microarray based detection of repetitive sequence changes. AB - Prior studies of oligonucleotide microarray-based mutational analysis have demonstrated excellent sensitivity and specificity except in circumstances where a frameshift mutation occurs in the context of a short repeated sequence. To further evaluate this circumstance, a series of nucleic acid samples having heterozygous mutations within repetitive BRCA1 sequence tracts was prepared and evaluated. These mutations included single nucleotide insertions and deletions in homopolymer runs, insertions and deletions of trinucleotide repeats, and duplications. Two-color comparative hybridization experiments were used wherein wild type reference and test targets are co-hybridized to microarrays designed to screen the entire BRCA1 coding sequence for all possible sequence changes. Mutations in simulated heterozygote samples were detected by observing relative losses of test target hybridization signal to select perfect match oligonucleotide probes. While heterozygous mutations could be readily distinguished above background noise in 9/19 cases, it was not possible to detect alterations in a poly dA/dT tract, small triplet repeat expansions, and a 10 bp direct repeat. Unexpectedly, samples containing (GAT)(3) triplet repeat expansions showed significantly higher affinity toward specific perfect match probes relative to their wild type counterparts. Therefore, markedly increased as well as decreased test sample hybridization to perfect match probes should be used to raise a suspicion of repetitive sequence changes. PMID- 11013447 TI - DNA 2000: International Symposium on the State-of-the-Art in Genetic Analysis, June 1-3, 2000, Boston, U.S.A. AB - DNA 2000, an international symposium on state-of-the-art genetic analysis, was held at the Back Bay Hilton in Boston, Massachusetts, on 1-3 June, 2000. Meeting highlights are described. The meeting was organized and sponsored by the California Separation Science Society (CaSSS; www.casss.org) and other co sponsors including the Human Genome Organisation (HUGO). DNA 2000 brought together a group of specialists in DNA detection and analysis methods from around the world in a venue presenting not only new technologies but also their applications in candidate gene studies, molecular diagnosis, analysis of complex diseases, and even studying the origin and evolution of humans. PMID- 11013448 TI - Allelic imbalance of BRCA1 transcript in the IVS20 12-bp insertion carrier. AB - One of the unclassified variants of the BRCA1 gene which has drawn considerable attention in recent years is the 12-bp insertion/duplication in intron 20. In this report, we show that a contribution from one chromosome cannot be detected in the BRCA1 transcript of the 12 bp insertion carrier. We also demonstrate here that the single transcript variant we observe by cDNA analysis originates from the same BRCA1 allele that harbours the 12-bp insertion. Hum Mutat 16:371, 2000. PMID- 11013449 TI - Factor IX mutations in South Africans and African Americans are compatible with primarily endogenous influences upon recent germline mutations. AB - Similar patterns of germline mutations in the factor IX gene (F9) have been observed in certain geographically and racially diverse populations. Germline mutation data have not been available from any region of Africa or from the Black race. Analysis of mutation data for Blacks is of interest, since this race has a high frequency of polymorphism compared to other races. This high frequency has been interpreted as evidence for the "out of Africa" hypothesis for the origin of humans, but it is possible that Blacks have a higher mutation rate due to genetic differences or environmental exposures. We report 26 independent mutations that were detected in patients of mixed races with hemophilia B from South Africa. The pattern of mutation in patients from this African country was similar to that of U.S. Caucasians. In addition, 22 independent mutation were detected in African American patients. The patterns of independent germline mutation in 22 African Americans (and in a combination 34 North American and African Blacks) is similar to that of U.S. Caucasians. Neither genetic differences between the Black and Caucasian races nor environmental and cultural differences between South Africa and the U.S. alter the germline pattern of mutation observed in F9. Hum Mutat 16:372, 2000. PMID- 11013450 TI - Homocystinuria in the Arab population of Israel: identification of two novel mutations using DGGE analysis. AB - This study describes, for the first time, a thorough genetic investigation in Israeli Arab homocystinuric patients. By using a DGGE methodology and sequencing we were able to identify the disease causing mutation in all. Of the mutations that were detected, two are novel: a 785C>G transversion in exon 7 (T262R) and a 5-bp deletion in the 5' of IVS17 including the T in the +2 position that is crucial for correct splicing (g18327-18331del5). In spite of the highly consanguineous nature of this population several different mutations were found. This may suggest that the mutations arose only recently in the population. The results of our study would enable early prenatal diagnosis, genetic counseling and screening for the mutations in population at risk. Hum Mutat 16:372, 2000. PMID- 11013451 TI - Molecular anatomy of CTG expansion in myotonin protein kinase gene among myotonic dystrophy patients from eastern India. AB - We have studied the CTG repeat sizes in the DMPK gene and six biallelic markers which are in complete linkage disequlibrium with Caucasian DM patients, to identify any common founder haplotype in 30 clinically diagnosed unrelated DM patients from eastern India. Our results revealed that in 27 patients (90%), CTG expansion took place on a DraIII(-) - HhaI(-) - Alu(+) - HinfI(+) - Fnu4H I(-) - TaqI(+) haplotype (haplotype I), similar to what have been published for Caucasoid and other DM patients. However, in three patients (10%), the expansion of CTG repeat was on DraIII(+) - HhaI(+) - Alu(+) - HinfI(-) - Fnu4H I(+) - TaqI( ) background (haplotype II), indicating a new haplotype. The distribution of haplotypes in 52 normal individuals of eastern India revealed that percentage of haplotypes I and II were 23.1% and 7.7% respectively in normal chromosomes. Haplotype II is absent among Caucasian DM patients as well as normal individuals indicating that this particular haplotype may be characteristic of the Indian population. Hum Mutat 16:372, 2000. PMID- 11013452 TI - Identification and characterization of two novel mutations that produce acute intermittent porphyria: A 3-base deletion (841-843delGGA) and a missense mutation (T35M). AB - A partial deficiency of Porphobilinogen deaminase (PBGD) is responsible for acute intermittent porphyria (AIP). AIP is inherited in an autosomal dominant fashion, and the prevalence in the Argentinean population is about 1:125,000. Here, two new mutations and two previously reported were found in the PBGD gene in 22 Argentinean AIP patients corresponding to 8 different families. To screen for AIP mutations in symptomatic patients, genomic DNA isolated was amplified in 6 PCR reactions, then all coding exons and flanking intronic regions were sequenced. The novel mutations are 841-843delGGA in exon 14, which results in the loss of glycine-281 (G281del), and one 104C>T point mutation in the exon 4 (T35M). To further characterize both novel mutations, the pKK-PBGD construct for the mutant alleles were expressed in E. coli, the enzymatic activity of the recombinant proteins were 1% and 4% of the mean level expressed by the normal allele for 841 843delGGA and T35M, respectively. Hum Mutat 16:373, 2000. PMID- 11013453 TI - Identification of a novel missense mutation L329I in the episodic ataxia type 1 gene KCNA1--a challenging problem. PMID- 11013454 TI - Homozygous familial hypercholesterolemia: A novel point mutation (W556R) in a Turkish patient. PMID- 11013455 TI - Five novel genetic variants in the promoter and coding region of the alpha B crystallin gene (CRYAB): -652G>A, -650C>G, -249G>C, S41Y, P51L. PMID- 11013456 TI - Two novel serine repeat length polymorphisms (1043insS and 1043insSS) at exon 23 of the TSC1 gene. PMID- 11013457 TI - A novel mutation (1653insC) in the thyroid hormone receptor beta in a patient resistant to thyroid hormone. PMID- 11013458 TI - Novel single nucleotide polymorphisms (3203A>G and 3204C>T) in the 3' end of the mitochondrial 16S rRNA gene. PMID- 11013459 TI - Metachromatic leukodystrophy: a novel mutation (c237delC) and extension of the haplotype associated with the P426L mutation. PMID- 11013460 TI - Identification of a novel polymorphism (IVS45+20 C/A) in the splice site of intron 45 of the ryanodine receptor gene (RYR1). PMID- 11013461 TI - Delta 469 mutation in the type 3 repeat calcium binding domain of cartilage oligomeric matrix protein (COMP) disrupts calcium binding. AB - Cartilage oligomeric matrix protein (COMP/TSP5), a large glycoprotein found in the territorial matrix surrounding chondrocytes, is the fifth member of the thrombospondin (TSP) gene family. While the function of COMP is unknown, its importance is underscored by the finding that mutations in the highly conserved type 3 repeat domain causes two skeletal dysplasias. Pseudoachondroplasia (PSACH) and Multiple Epiphyseal Dysplasia, Fairbanks type (EDM1). The type 3 repeats are highly conserved low-affinity Ca(2+)binding domains that are found in all TSP genes. This study was undertaken to determine the effects of mutations on calcium binding and structure of the type 3 repeat domains. Wild-type (WT) and Delta469 recombinant COMP (rCOMP) proteins containing the entire calcium-binding domain were expressed in E. coli and purified. Equilibrium dialysis demonstrated that WT bound 10-12 Ca(2+)ions/molecule while Delta469 bound approximately half the Ca(2+)ions. Circular dichroism (CD) spectrometry had striking spectral changes for the WT in response to increasing concentrations of Ca(2+). These CD spectral changes were cooperative and reversible. In contrast, a large CD spectral change was not observed at any Ca(2+)concentration for Delta469. Moreover, both WT and Delta469 proteins produced similar CD spectral changes when titrated with Zn(2+), Cu(2+)and Ni(2+)indicating that the Delta469 mutation specifically affects only calcium binding. These results suggest that the Delta469 mutation, in the type 3 repeat region, interferes with Ca(2+)binding and that filling of all Ca(2+)binding loops may be critical for correct COMP protein conformation. PMID- 11013462 TI - Selective down-regulation of IP(3)receptor subtypes by caspases and calpain during TNF alpha -induced apoptosis of human T-lymphoma cells. AB - There are at least three types of inositol 1,4,5-trisphosphate receptor (IP(3)R) [IP(3)-gated Ca(2+)channels], which are expressed in different cell types and mammalian tissues. In this study, we have identified three IP(3)R subtypes in human Jurkat T-lymphoma cells. All three subtypes have a molecular mass of about 260 kDa, and display Ca(2+)channel properties in an IP(3)-dependent manner. We have also demonstrated that TNFalpha promotes the activity of different proteases (e.g. caspase-8, caspase-3 and calpain), alters the TCR-mediated Ca(2+)response and subsequently induces apoptosis in Jurkat cells. During the first 6 h of incubation with TNFalpha, several IP(3)R subtype-related changes occur (e.g. proteolysis of IP(3)R subtypes, inhibition of IP(3)binding and impairment of IP(3)-mediated Ca(2+)flux) concomitantly with an elevation of protease (caspase 8, caspase-3 and calpain) activity. Furthermore, the caspase inhibitor, Z-VAD fmk, significantly reduces TNFalpha-mediated perturbation of IP(3)R1 and IP(3)R2 (but not IP(3)R3) function; whereas the calpain inhibitor I, ALLN, is capable of blocking the inhibitory effect of TNFalpha on IP(3)R3 function. These findings suggest that IP(3)R1 and IP(3)R2 serve as cellular substrates for caspases, and IP(3)R3 is a substrate for calpain. We propose that the selective down-regulation of IP(3)R subtype-mediated Ca(2+)function by caspase-dependent and calpain sensitive mechanisms may be responsible for the early onset of the apoptotic signal by TNFalpha in human T-cells. PMID- 11013463 TI - NO-induced modulation of calcium-oscillations in pulmonary vascular smooth muscle. AB - The effect of the nitric oxide (NO) donor sodium nitroprusside (SNP) on both [Ca(2+)](i)and mechanical activity was studied in the rat isolated pulmonary artery (RPA). In freshly isolated myocytes loaded with 1 microM indo lacetoxymethyl ester for 30 min, short (40-60 s) application of ATP (100 microM) or ET-1 (0.1 microM) induced 3-6 cyclic rises in [Ca(2+)](i)(Ca-oscillations) of decreasing amplitude. Preincubation of cells with SNP (10-250 microM) for 10 min had no effect on the resting [Ca(2+)](i)value, but progressively abolished the oscillations. A similar effect was obtained with 8-bromo-cGMP (100-500 microM). SNP (0.001-100 microM) concentration-dependently relaxed ATP (10 mM, n = 4) and ET-1 (0.1 microM, n = 4)-precontracted RPA. 1H-[1,2,4]oxadiazolol [4,3, a]quinoxalin-1-one (ODQ, 10 microM), a potent inhibitor of the cytosolic guanylyl cyclase, fully reversed the effect of SNP on ATP-induced [Ca(2+)](i)oscillations as well as on ATP-precontracted RPA. In contrast, N-[2-(methylamino)ethyl]-5 isoquinolinesulfonamide (H8, 10 microM), a potent inhibitor of cGMP-dependent protein kinase (PKG), did not alter the effect of SNP. Caffeine (5 mM) induced only one transient [Ca(2+)](i)-increase (n = 24), the amplitude of which was altered neither by SNP nor by 8-bromo-cGMP. Our results show that the relaxing effect of NO in RPA is related, at least in part, to its action on the Ca signalling pathway. NO interacts with inositol trisphosphate pathway without interacting with the ryanodine-sensitive receptor. Finally, the effect of NO involves an increase in cGMP but appears independent of activation of PKG. PMID- 11013465 TI - Dynamic ca(2+)changes in neutrophil phagosomes A source for intracellular ca(2+)during phagolysosome formation? AB - An increase in cytosolic Ca(2+)concentration periphagosomally is critical for phagolysosomal formation and neutrophil elimination of microbes. The Ca(2+)increase could be achieved through release of Ca(2+)from mobilized intracellular stores. Alternatively, Ca(2+)that passively enter the phagosome during phagocytosis could be provided by the phagosome. Intraphagosomal Ca(2+)changes in single human neutrophils was measured during phagocytosis of serum opsonized Fura-2-conjugated zymosan particles, using a digital image processing system for microspectrofluorometry. A decrease in phagosomal Ca(2+)down to nanomolar concentrations was seen within minutes following phagosomal closure. Blockage of plasma membrane Ca(2+)channels by econazole abolished this decrease. The fluorescence properties of Fura-2 zymosan were retained after phagocytosis and stable to pH changes, reactive oxygen species, and proteolytic enzymes. We suggest that Ca(2+)ions present in the phagosome enter the cell cytosol through Ca(2+)channels in the phagosomal membrane, achieving a localized Ca(2+)rise that is important for phagosome processing. PMID- 11013464 TI - Calcium signalling in sarcoplasmic reticulum, cytoplasm and mitochondria during activation of rabbit aorta myocytes. AB - This study investigated the relationship between cytoplasmic, mitochondrial, and sarcoplasmic reticulum (SR) [Ca(2+)] in rabbit aorta smooth muscle cells, following cell activation. Smooth muscle cells were loaded with the Ca(2+) sensitive fluorescent indicator Mag-Fura-2-AM, and then either permeabilized by exposure to saponin, or dialyzed with a patch pipette in the whole-cell configuration to remove cytoplasmic indicator. When the intracellular solution contained millimolar EGTA or BAPTA, activation of SR Ca(2+)release through IP(3)or ryanodine receptors induced a decrease in the [Ca(2+)] reported by Mag Fura-2. However, when EGTA was present at < or =100 microM, the same stimuli caused an increase in the [Ca(2+)] reported by Mag-Fura-2. The increase in [Ca(2+)] caused by phenylephrine or caffeine was delayed, and prolonged, with respect to the cytoplasmic Ca(2+)transient. Evidence is presented that this Mag Fura-2 signal reflected a rise in mitochondrial [Ca(2+)]. Agents that inhibit mitochondrial function, such as FCCP or cyanide in combination with oligomycin B, converted the increase in organelle Mag-Fura-2 fluorescence to a decrease, while also prolonging the cytoplasmic Ca(2+)transient. There was considerable similarity between the localization of Mag-Fura-2 fluorescence and the mitochondria-selective indicator tetramethylrhodamine ethyl ester. Thus, we propose that there is close functional integration between the SR and mitochondria in aorta smooth muscle cells, with mitochondria taking up Ca(2+)from the cytoplasm following cell activation. PMID- 11013466 TI - Progress in ca(2+)dynamics: small temporal and spatial scales drive macroscopic dynamics. PMID- 11013467 TI - Duplicate and salami publications. PMID- 11013469 TI - The palm print as a sensitive predictor of difficult laryngoscopy in diabetics: a comparison with other airway evaluation indices. AB - AIMS: To evaluate the ink impression made by the palm of the dominant hand as a screening tool for difficult laryngoscopy in diabetic patients. SUBJECTS AND METHODS: In this prospective study, airway of 50 adult diabetic patients, undergoing elective surgery under general anaesthesia, was assessed preoperatively using the common clinical indices such as Modified Mallampati test, thyromental distance, degree of head extension and a specific index- the palm print test. Following induction of anaesthesia and neuromuscular relaxation, laryngoscopy was performed and the laryngoscopic view scored. The sensitivity, specificity and positive predictive value of each airway evaluation index were calculated. RESULTS: The incidence of difficult laryngoscopy was 16%. The palm print test had the highest sensitivity (75%) of all the indices. The thyromental distance less than six cm had the highest specificity (95.2%) but was least sensitive (25%). 87% of patients with difficult laryngoscopy had two or more indices abnormal. CONCLUSION: Though the palm print test was the most sensitive index of the four indices studied, a better prediction of difficult laryngoscopy can be achieved by evaluating all the four airway indices preoperatively. PMID- 11013468 TI - The evaluation of cerebral oxygenation by oximetry in patients with ischaemic stroke. AB - AIMS: To evaluate the clinical significance of estimation of the regional cerebral oxygen saturation (rSO2) in the patients with ischaemic stroke by the cerebral oximetry during acute, sub-acute and chronic phases. SUBJECTS AND METHODS: In this prospective study, 24 patients with ischaemic stroke in the middle cerebral artery territory were included. A detailed clinical examination and appropriate laboratory investigations were carried out. The rSO2 was determined by oximetery (INVOS 3100-SD) bilaterally on the first, third, seventh, and fifteenth days. The blood pressure, the peripheral capillary oxygen saturation and the arterial blood gas values were noted too. the changes were evaluated along with Glasgow coma scale (GCS) using unpaired student t-test and one way ANOVA test. RESULTS: There were significant differences between the rSO2 values in acute, subacute and chronic phases on the side of the lesion (p value < 0.05). The values of oxygen saturation gradually increased throughout the chronic phase. These values showed a positive correlation with GCS, but the results were not significant statistically. The rSO2 values were also significantly higher on the non-lesional side than those on the lesion side in the acute phase (p= 0.0034), the discrepancy disappeared during the sub-acute and chronic phases. CONCLUSION: Cerebral oximetry can be used as a measure to evaluate the cerebral oxygenation during the various phases of ischaemic stroke. It has a potential to serve as a useful marker for detection of cerebral oxygenation imbalances, to judge the effectiveness of the management and for the follow-up of patients with ischaemic stroke. PMID- 11013470 TI - Giant prosthesis for reinforcement of visceral sac for complex bilateral and recurrent inguinal hernias: a prospective evaluation. AB - AIMS: To evaluate giant prosthesis for reinforcement of visceral sac (GPRVS) as a treatment for complex bilateral and recurrent inguinal hernias. SUBJECTS AND METHODS: The prospective study carried out in a single surgical unit at a tertiary health care center involved consecutive series of 31 patients with complex bilateral and recurrent inguinal hernias who underwent GPRVS. All were men and the mean age was 58 years (range 49-95 years). Factors predicting high risk for recurrence included a large hernia ( greater, similar5cms, 32%, 10/31 patients), failure of one or more previous repairs (45%, 14/31 patients), chronic obstructive pulmonary disease (25%, 8/31 patients) and poor muscle tone (70%, 22/31 patients). Operative time, length of postoperative stay, complications and death were the main outcome measures. RESULTS: Mean -/+ SEM operative time was 65 -/+ 11 minutes (range 45-115 minutes). Mean -/+ SEM length of stay was 3.5 -/+ 0.7 days (range 2-5 days). There were 4 minor complications, but no mesh infections and death. Follow up was obtained for a mean period of 14.6 months (range 12-23 months); there were no recurrences. CONCLUSION: GPRVS provides a definitive and safe cure for repair of complex bilateral and recurrent inguinal hernias because of its simplicity, ease of the procedure, good results and low recurrence rate. PMID- 11013471 TI - A study of an epidemic of acute respiratory disease in Jaipur town. AB - AIM: To detect an association between the sudden epidemic with respiratory symptoms, and fogging with dichlorovos in Jaipur town and to find out probable mechanism of causation of the epidemic. SUBJECTS AND METHODS: In this community based study of the epidemic, house to house survey of households selected using systematic random sampling was carried out. The incidence in the exposed and unexposed population, the relative risk and attributable risk were calculated. RESULTS: The incidence of cases was high (58.9%) in subjects present on roads at the time of fogging as compared to in those who were inside rooms of the houses (5.4%) and in those who were not in the locality at that time (1.8%) [Relative Risk (RR)=32.7 and Attributable Risk (AR)=96.9%]. CONCLUSION: High RR and AR in the present epidemic indicate strong association between fogging and occurrence of symptoms. In absence of signs and symptoms of organophosphorus poisoning it suggests that this could have been due to an inappropriate solvent or defective functioning of fog generator, leading to generation of an unusual dark fog, that might have irritated eyes and respiratory tract of exposed residents. PMID- 11013472 TI - Scleredema adultorum. AB - Scleredema adultorum is a rare connective tissue disorder reported usually following streptococcal infection, influenza, measles, and mumps. It has been reported occasionally following trauma and tuberculous lymphadenitis. This is a report of scleredema adultorum developing after chicken pox in an eight-year-old male child. The diagnosis was established by characteristic picture on skin biopsy using special stain. The patient had a benign course and a spontaneous recovery in two weeks. The case has been reported as the first case of scleredema adultorum developing after chicken pox. PMID- 11013473 TI - An intragastric trichobezoar: computerised tomographic appearance. AB - A 26-year-old lady presented with a history of abdominal pain and distension since two months. The ultrasound examination showed an epigastric mass, which was delineated as a filling defect in the stomach on barium studies. The computerised tomographic scan showed a gastric mass with pockets of air in it, without post contrast enhancement. This case highlights the characteristic appearance on computerised tomography of a bezoar within the stomach, a feature that is not commonly described in medical literature. PMID- 11013474 TI - Sprengels deformity: anaesthesia management. AB - A 28 years old lady presented with Sprengels deformity and hemivertebrae for Fothergills surgery. Clinically there were no anomalies of the nervous, renal or the cardiovascular systems. She had a short neck and score on modified Mallapati test was grade 2. She was successfully anaesthetised using injection Propofol as a total intravenous anaesthetic agent after adequate premedication with injection Midazolam and injection Pentazocine. Patient had an uneventful intraoperative and postoperative course. PMID- 11013475 TI - Neurofibromatosis-Noonan syndrome or LEOPARD Syndrome? A clinical dilemma. AB - Neurofibromatosis (NF), Noonan syndrome (NS), and LEOPARD syndrome are all autosomal dominant conditions, each being a distinct clinical entity by itself. Rarely, one encounters cases with features of NF and NS and is termed as the 'Neurofibromatosis-Noonan syndrome' (NF-NS). The authors report a clinical dilemma with major clinical features of the NF-NS syndrome and LEOPARD syndrome co-existing in the same patient. Also, features of Noonan syndrome and LEOPARD syndrome are compared with the case reported. PMID- 11013476 TI - Eisenmenger syndrome in pregnancy. AB - Maternal mortality in the presence of Eisenmenger syndrome is reported to be 30 to 50% & increases further with associated complications. A case of Eisenmenger syndrome in pregnancy where the patient progressively deteriorated postpartum & expired 3 weeks later is reported. PMID- 11013477 TI - Imaging findings in a giant hepatic artery aneurysm. AB - A rare case of relatively asymptomatic giant hepatic artery aneurysm of atherosclerotic aetiology is presented. The importance of imaging findings in the diagnosis of this condition and the differential diagnosis including the pertinent literature on the topic is discussed. PMID- 11013478 TI - Malrotation of the gut manifested during pregnancy. AB - The diagnosis of intestinal obstruction during pregnancy poses problems, as vomiting which is an important symptom of the obstruction can be attributed to hyperemesis of pregnancy and radiological investigation are avoided during this period. A case of intestinal obstruction due to volvulus resulting from congenital malrotation of the gut is reported here. The patient first presented during pregnancy. The case emphasises the need for thorough investigations in a case of persistent vomiting in pregnancy. PMID- 11013479 TI - Isolated tuberculous hepatic abscess in a non-immunocompromised patient. AB - A 38 years old female presented with pain in the epigastrium, jaundice and fever since one and half month. The computerised tomographic scan of the abdomen revealed a multiloculated abscess of the left lobe of liver. The pus drained from the liver abscess at laparotomy showed acid fast bacilli on microscopy. A detailed search failed to identify any other focus of tuberculous infection. The case has been reported for the rarity of isolated hepatic tuberculous abscess and its presentation with jaundice, a rare feature, and to highlight the importance of microscopic or culture diagnosis in a suspected case of pyaemic abscess. PMID- 11013480 TI - Giant cell tumour of talar body. AB - Giant cell tumour (osteoclastoma) of talar bone is a rare entity and is seen more commonly in the third decade of life. We report this disease entity in a 17-years old girl. The patient presented with painful swelling of the left ankle with an osteolytic lesion in the talus on conventional radiographs. Intralesional curettage and autologous bone grafting was performed following which patient's pain and swelling disappeared. Complete range of movement at the ankle joint was regained with minimal restriction at the subtalar joint. There is no evidence of relapse at six months follow up. PMID- 11013481 TI - The art and science of presentation: the poster. PMID- 11013482 TI - Information retrieval in medicine: overview and applications. PMID- 11013483 TI - The art and science of web-based literature search: the MEDLINE. PMID- 11013484 TI - Images in medicine: Apert syndrome. PMID- 11013487 TI - Non-pathology: the bedrock of pathology. AB - Pathology, also called morbid anatomy, is macroscopically, microscopically, and molecularly so manifest an array of phenomena that it has compelled medical men to closely link it up with disease, dis-ease, and death. But there is more than meets the eye of the morbid anatomists, microscopists, and the molecular biologists. The obvious science of pathology is governed by numerous abstract, subtle, non-pathological factors. A pathological phenomenon is subservient to cosmic noumenon. Such a sea-change allows a newer perspective that cures modern medicine of many of its dogmas and provides epistemologically valid directions to research methodologies on the one hand and clinical practices on the other. PMID- 11013485 TI - Images in radiology: type III growing skull fracture. PMID- 11013486 TI - Images in pathology: verrucous haemangioma. PMID- 11013488 TI - Antinuclear antibodies: clinical applications. AB - One of the common serological hallmarks of autoimmune disorders is the presence of various autoantibodies in the sera of patients affected by these disorders. Antinuclear antibodies (ANA) detection is often needed to aid the diagnosis in several autoimmune disorders. In view of the different methodologies available for their detection, it becomes essential to understand the advantages and pitfalls of each procedure. This brief review discusses some methodological aspects of ANA detection and the clinical relevance of the presence of some of the autoantibodies found in the sera of patients with autoimmune disorders. PMID- 11013489 TI - Non-invasive respiratory monitoring in paediatric intensive care unit. AB - Monitoring respiratory function is important in a Paediatrics Intensive Care Unit (PICU), as majority of patients have cardio-respiratory problems. Non-invasive monitoring is convenient, accurate, and has minimal complications. Along with clinical monitoring, oxygen saturation using pulse oximetry, transcutaneous oxygenation (PtcO2) and transcutaneous PCO2 (PtcCO2) using transcutaneous monitors and end-tidal CO2 using capnography are important and routine measurements done in most PICUs. Considering the financial and maintenance constraints pulse oximetry with end tidal CO2 monitoring can be considered as most feasible. PMID- 11013491 TI - Imagery PMID- 11013490 TI - Eptifibatide: in the treatment of acute coronary syndromes. PMID- 11013492 TI - Fos expression in the rat brain and spinal cord evoked by noxious stimulation to low back muscle and skin. AB - STUDY DESIGN: Acute noxious stimulation delivered to lumbar muscles and skin of rats was used to study Fos expression patterns in the brain and spinal cord. OBJECTIVES: The present study was conducted to determine the differences in Fos expression in the brain and spinal cord as evoked by stimuli delivered to lumbar muscles and skin in rats. SUMMARY OF BACKGROUND DATA: Patients with low back pain sometimes show psychological symptoms, such as quiescence, loss of interest, decreased activities, appetite loss, and restlessness. The pathway of deep somatic pain to the brain has been reported to be different from that of cutaneous pain. However, Fos expression has not been studied in the central nervous systems after stimulation of low back muscles. METHODS: Rats were injected with 100 L of 5% formalin into the multifidus muscle (deep pain group; n = 10) and into the back skin of the L5 dermatome (cutaneous pain group; n = 10). Two hours after injection, the distribution of Fos-immunoreactive neurons was studied in the brain and spinal cord. RESULTS: Fos-immunoreactive neurons were observed in laminae I-V in the spinal cord in the cutaneous pain group, but they were not seen in lamina II in the deep pain group. In the brain, Fos immunoreactive neurons were significantly more numerous in the deep pain group than in the cutaneous pain group in the piriform cortex, the accumbens nucleus core, the basolateral nucleus of amygdala, the paraventricular hypothalamic nucleus, the ventral tegmental area, and the ventrolateral periaqueductal gray. CONCLUSION: The finding that Fos-immunoreactive neurons were absent from lamina II of the spinal cord in the deep pain group is similar to that of the projection pattern of the visceral pain pathway. Fos expression in the ventrolateral periaqueductal gray in the deep pain group may represent a reaction of quiescence and a loss of interest, activities, or appetite. Furthermore, the detection of large numbers of Fos-immunoreactive neurons in the core of accumbens nucleus, basolateral nucleus of amygdala, paraventricular hypothalamic nucleus, and ventral tegmental area in the deep pain group may suggest a dominant reaction of dopaminergic neurons to stress, and a different information processing pathway than from that of cutaneous pain. PMID- 11013493 TI - Impact response of the intervertebral disc in a finite-element model. AB - STUDY DESIGN: A three-dimensional nonlinear poroelastic finite-element model of a vertebra disc was used to analyze the biomechanical effects of impact loading on the spinal segment. OBJECTIVES: To predict changes in biomechanical parameters such as intradiscal pressure, dynamic stiffness, stresses in the endplate region, and the shock-absorbing mechanism of the spine under different impact duration/loading rates, and to investigate the relation between the rate of loading and the fracture potential of the vertebral body. SUMMARY OF BACKGROUND DATA: It is not practical to discern the role of impact duration using experimental protocols. Analytical studies are better suited to this purpose. However, previous poroelastic finite-element models of the motion segments have dealt mostly with creep phenomena. METHODS: A three-dimensional, L3-L4 motion segment, finite-element model was modified to incorporate the poroelastic properties of the disc, endplate, and cancellous core, and thus simulate the shock-absorbing phenomena. The results were analyzed under variable impact durations for a constant maximum compressive impact load of 3 kN. RESULTS: For a shorter impact duration and a given F(max), relatively high cancellous core pressure was generated as compared with a case of long impact duration, although the amount of impulse was increased. In contrast, relatively constant pore pressures were generated in the nucleus regardless of the impact duration. The changes in spinal segment stiffness as a function of impact duration indicated that for a shorter duration of impact, high dynamic stiffness increases the stability of the spinal segment against the impact load. However, the corresponding increase in stresses within the vertebral body and endplate may produce fractures. CONCLUSIONS: The finite-element technique was used to address the role of impact duration in producing trauma to the spinal motion segment. Within the limitations of the model, the results suggest that fractures are likely to occur under shorter impact duration conditions. Depending on the strength of the region, a fracture may be initiated in the endplate region or the posterior wall of the cortical shell. The nucleus pressure is independent of the impact duration and depends only on the magnitude of the impact force. PMID- 11013494 TI - Patient characteristics and patterns of use for lumbar spine radiographs: results from the Veterans Health Study. AB - STUDY DESIGN: Longitudinal data from the Veterans Health Study, an observational study of male patients receiving Veterans Administration ambulatory care, were analyzed. OBJECTIVE: To identify patient characteristics that predict different patterns in the use of lumbar spine radiographs. SUMMARY AND BACKGROUND DATA: In this study, 401 patients with low back pain receiving ambulatory care services in four Veterans Administration outpatient clinics in the greater Boston area were followed for 12 months. METHODS: Participants were mailed the Medical Outcome Study Short Form Health Survey and participated in scheduled interviews that included the completion of a low back questionnaire, a comorbidity index, and a straight leg raising test. Four groups of patients were defined according to the patterns of use for lumbar spine radiographs: prior use, repeat use, no use, and new use of lumbar spine radiographs. These groups were compared in terms of sociodemographics, comorbid conditions, low back pain intensity, radiating leg pain, straight leg raising, Medical Outcome Study Short Form Health Survey scores, and low back disability days. RESULTS: The patients with new lumbar spine radiographs showed worse physical and psychological distress than the participants in the other three groups. In contrast, the patients with no lumbar spine radiographs reported minor physical impairment. Compared with patients who had no repeat radiographs, patients with repeat lumbar spine radiographs had similar scores on physical health, but they showed worse scores of mental health. CONCLUSIONS: Both physical and psychological factors contribute to having new radiographic examinations, whereas psychological factors have increased importance in the repeat use of roentgenographic examinations. Repeat radiographs appear to be overused, judging by the severity of physical impairment as measured by low back pain intensity, the Medical OutcomeStudy Short Form Health Survey, and disability days. PMID- 11013495 TI - Patient-oriented outcomes from low back surgery: a community-based study. AB - STUDY DESIGN: This study used a prospective cohort design. OBJECTIVE: To examine factors associated with favorable self-reported patient outcomes 1 year after elective surgery for degenerative back problems. SUMMARY OF BACKGROUND DATA: Many previous studies addressing the results of low back surgery have been conducted in academic institutions or by single surgeons. As part of a quality improvement effort, surgeons in private practice led a community-based outcomes management project in Washington State. METHODS: Patients ages 18 and older with the following diagnoses were eligible for the study: degenerative changes, herniated disc, instability, and spinal stenosis. Nine orthopedists and neurosurgeons enrolled a total of 281 patients. Participants were asked to complete baseline and 1-year follow-up surveys. Data concerning diagnoses, clinical signs, and operative procedures were provided by the surgeons. The researchers examined sociodemographic characteristics, self-reported symptoms before surgery, preoperative clinical signs, diagnoses, and operative procedures associated with three primary outcomes: better functioning, improved quality of life, and overall treatment satisfaction. RESULTS: Follow-up surveys were completed by 236 (84%) of the enrolled patients. Approximately two thirds of the study participants reported much better functioning (65%), a great quality of life improvement (64%), and a very positive perspective about their treatment outcome (68%). The following variables were associated with worse patient outcomes: older age, previous low back surgery, workers' compensation coverage, and consultation with an attorney before surgery. Patients undergoing a fusion procedure were more likely to report good outcomes. CONCLUSIONS: The authors' experience indicates that community-based outcomes data collection efforts are feasible and can be incorporated into usual clinical practice. The study results indicate that compensation payments and litigation are two important predictors of poor outcomes after low back surgery in community practice. Because of small numbers, varied diagnoses, and possible selection bias, the findings with respect to fusion should be interpreted cautiously. PMID- 11013496 TI - Loss of sagittal plane correction after removal of spinal implants. AB - STUDY DESIGN: A retrospective review of a clinical series was performed. OBJECTIVES: To evaluate the incidence of adult patients who experienced spinal collapse after spinal implant removal after a long spinal arthrodesis, and to assess the various factors that may influence the likelihood of collapse after implant removal. SUMMARY OF BACKGROUND DATA: Published reports describing the benefits or complications of spinal implant removal do not exist. Spinal implant removal, often considered a benign procedure, is even required by the Food and Drug Administration (FDA) for certain implants. METHODS: The medical records and radiographs of 116 consecutive adult patients with long posterior instrumented fusions (>5 segments) were reviewed. The information obtained included original diagnosis, patient age, number of previous surgeries before implant removal, levels of anterior and posterior fusion, time from fusion to implant removal, time from implant removal to failure, and reason for hardware removal. Radiographs also were assessed including scoliosis, lordosis, and kyphosis measurements before implant removal, after hardware removal, after failure, and after revision surgery. RESULTS: Of 116 patients, 14 underwent spinal implant removal. Most of these patients reported prominent implants either proximally in the thoracic spine or distally in the ilium (Galveston technique). Of these 14 patients, 4 experienced increased pain and collapse after implant removal despite thorough intraoperative explorations demonstrating solid fusion. CONCLUSIONS: Spinal implant removal after long posterior fusion in adults may lead to spinal collapse and further surgery. Removal of instrumentation should be avoided or should involve partial removal of the prominent implant. PMID- 11013497 TI - Deep wound infections after neuromuscular scoliosis surgery: a multicenter study of risk factors and treatment outcomes. AB - STUDY DESIGN: A retrospective case-control study evaluating risk factors for infection, causative organisms, and results of treatment in patients with cerebral palsy or myelomeningocele who underwent fusion for scoliosis was performed. OBJECTIVES: To identify risk factors for infection, and to characterize the infections in terms of infecting organisms and response to treatment. SUMMARY OF BACKGROUND DATA: No previous studies have analyzed risk factors or causative organisms, nor have they indicated results of treatment for infections in this group of patients. METHODS: After a 10-year retrospective review of 210 surgically treated patients, deep wound infections developed in 16 patients with myelomeningocele and 9 patients with cerebral palsy. These patients were studied extensively for possible risk factors, along with 50 uninfected patients matched for age, diagnosis, and year of surgery. Statistical testing was performed to identify risk factors. The courses of the infections were characterized in terms of organisms isolated and response to treatment. Treatment was performed in a stepwise fashion and classified in terms of the most successful step: debridement and closure, granulation over rods, or instrumentation removal. RESULTS: Of the 10 risk factors tested, 2 were found to be significant: degree of cognitive impairment and use of allograft. Findings showed that 52% of the infections were polymicrobial. Gram-negative organisms were isolated as commonly as gram-positive organisms. The most common organisms were coagulase-negative Staphylococcus, Enterobacter, Enterococcus, and Escherichia coli.- Debridement and closure were successful in 11 of 25 patients with deep wound infection. Of the 14 patients with infection not resolved by serial debridements and closure, 2 were managed successfully by allowing the wound to granulate over rods, and 7 required rod removal for persistent wound drainage. There were three symptomatic pseudarthroses. Infections resulting from gram-positive organisms were most often managed successfully with debridement and closure (P = 0.012). CONCLUSIONS: Patients with cerebral palsy or myelomeningocele who have severe cognitive impairment, and those who received allograft may be at increased risk for infection. Infections are more often polymicrobial and caused by gram-negative organisms than is typical for elective orthopedic procedures. This suggests an enteric source. Treatment with debridement and closure was not always successful. Patients in whom infection develops are then at increased risk for pseudarthrosis. PMID- 11013498 TI - Surface electrode somatosensory-evoked potentials in spinal surgery: implications for indications and practice. AB - STUDY DESIGN: A retrospective study of 442 major spinal operations with spinal cord monitoring performed in a University Hospital between 1982 and 1992 was performed. OBJECTIVES: To assess the reliability of the authors' method for monitoring by somatosensory-evoked potential recording, to determine criteria for intraoperative corrective action, and to redefine the need for the wake-up test. SUMMARY OF BACKGROUND DATA: The routine use of somatosensory-evoked potential monitoring in spinal surgery remains controversial. In Nottingham, the authors have used a method of recording from either scalp or high cervical electrodes. METHODS: The recordings and outcomes of all monitored spinal operations between the years 1982 and 1992 were reviewed. RESULTS: In 442 procedures, 23 technical failures (no reliable monitoring) occurred. Most technical failures were in patients with severe preoperative neurology, identifiable by somatosensory-evoked potential recording before operation. In the remaining 419 procedures, a significant intraoperative change in response occurred in 70 procedures (16.7%). Using the definitions of the American EEG Society, the authors identified 10 true positives and 60 false-positives. There were no false-negatives. A wake-up test was performed if an amplitude drop greater than 50% from baseline value persisted after attempts to correct any possible identifiable intraoperative cause. This occurred in only 21 patients (5%). In the true-positive group, somatosensory evoked potential recordings remained persistently abnormal despite an apparently normal subsequent wake-up test. The sensitivity of the method according to current definitions was 100% and the specificity 85.33%. CONCLUSIONS: Modified guidelines are needed for routine intraoperative use of somatosensory-evoked potential monitoring in spinal surgery. Such guidelines should avoid the term "false-positive" as currently defined and concentrate on the causative analysis of abnormal responses that warn of critical spinal cord dysfunction before that becomes irreversible and allow for appropriate action. Information from this monitoring method alerted the surgeon to the possible need for corrective action in an additional 9.78% of the reported patients, who traditionally would have been regarded as false-positives. A wake-up test still is indicated in patients with persistent suppression of their somatosensory-evoked potential despite correction of any identifiable cause and in cases of technical failure. The current method proved flexible, versatile, and reliable for future use. PMID- 11013499 TI - One-year prevalence of low back pain in two Swiss regions: estimates from the population participating in the 1992-1993 MONICA project. AB - STUDY DESIGN: A cross-sectional survey was performed. OBJECTIVE: To estimate the extent of low back pain as a public health problem. SUMMARY OF BACKGROUND DATA: Health surveys converge on very high estimates of low back pain in general populations, but few studies have included severity criteria in their definition and conclusions. Because it is unlikely that interventions will influence the prevalence of minimal and infrequent symptoms, greater attention should be paid to characteristics of low back pain that indicate some impact on the life of survey respondents. METHODS: Two regions participated in the MONICA (MONitoring of trends and determinants in CArdiovascular disease) project in Switzerland. Participants randomly selected from the general population completed a standard self-administered questionnaire on cardiovascular risk factors. A special section on low back pain was added in the third (1992-1993) MONICA survey and completed by 3227 participants. RESULTS: A regional difference found in the 12-month prevalence rate disappeared with the inclusion of severity criteria. Low back pain over more than seven cumulated days was reported among men by 20.2% (age range, 25-34 years) to 28.5% (age range, 65-74 years), respectively, among women by 31.1% to 38.5%. Similar rates of reduction in activity (professional, housekeeping, and leisure time) and medical consultation (conventional and nonconventional) motivated by low back pain characterized the two participating regions. The cumulative duration of pain was related to all the indicators showing the impact of low back pain on everyday life. CONCLUSIONS: Determining the cumulative duration of low back pain over the preceding year is a straightforward task, and a cutoff at 1 week seems appropriate for distinguishing between low- and high-impact low back pain. PMID- 11013500 TI - Socioeconomic factors and disability retirement from back pain: a 1983-1993 population-based prospective study in Norway. AB - STUDY DESIGN: A prospective observational study with an 11-year follow-up period was performed. OBJECTIVE: To investigate the influence of education and socioeconomic position on the incidence of permanent disability retirement from back pain. SUMMARY OF BACKGROUND DATA: Early retirement because of back pain is the extreme end point of a disabling process that is a great burden to the individual and costly for the society. Groups of employees at particular risk for permanent back pain disability need to be identified. METHODS: All employed men and women in Norway between the ages of 20 and 53 years in 1980 were included (n = 1,333,556). Outcome measures were disability retirement from inflammatory back pain (ICD-9 code 720) and noninflammatory back pain (ICD-9 codes 721 to 724). RESULTS: The 11-year cumulative incidence was 0.15% (n = 1990) for disability retirement from inflammatory back pain and 1. 64% (n = 21,829) for noninflammatory back pain and was somewhat higher in women than in men. Each year of formal education was independently associated with decreased risk for disability retirement from noninflammatory back pain (odds ratio [OR] = 0.78; 95% confidence interval [CI] = 0.77-0.79) and from inflammatory back pain (OR = 0.83; 95% CI = 0.81-0.86). Whereas disability from inflammatory back pain was moderately associated with socioeconomic status, there was a consistent upward trend in the association between disability retirement from noninflammatory back pain and lower socioeconomic position. The OR for unskilled workers was 3.1 (95% CI = 2.6-3.7) for men and 2.1 (95% CI = 1.7-2.5) for women, as compared with that of higher professionals. Stepwise analyses suggest that the effect of education is not mediated by socioeconomic status. CONCLUSIONS: The consistent upward trend in the relation of disability retirement to lower levels of education and socioeconomic position, even for inflammatory back pain, shows that factors related to the occupational and social environment play an important role in the disabling process. The stepwise, monotonic relation between socioeconomic position and disability retirement from back pain, even at the higher end of the socioeconomic scale, suggests that the relation between social class and back pain disability cannot be explained solely in terms of manual versus nonmanual jobs. PMID- 11013501 TI - Differences in repositioning error among patients with low back pain compared with control subjects. AB - STUDY DESIGN: Trunk repositioning error was measured in 20 patients with chronic low back pain and 20 control subjects. OBJECTIVES: To measure trunk repositioning error as a method of measuring proprioception of the low back and to compare trunk repositioning error in patients with low back pain and in control subjects. SUMMARY OF BACKGROUND DATA: Although many current low back pain rehabilitation programs incorporate proprioceptive training, very little research has been performed on proprioception of the low back. METHODS: While standing with the legs and pelvis immobilized, the subject bent the trunk to a predetermined target position and then attempted to replicate the position. Repositioning error was calculated as the absolute difference between the actual target position and the subject-perceived target position. The multiple target positions in the frontal and sagittal planes were tested. Trunk position was measured with a 3Space Tracker, which analyzes the three-dimensional position of the body. RESULTS: Repositioning error in patients with low back pain was significantly higher than that of control subjects in flexion, and significantly lower than that of control subjects in extension. CONCLUSIONS: The increase in repositioning error of patients with low back pain during flexion implies that some aspects of proprioception are lost in patients with low back pain. The decrease in repositioning error in patients with low back pain in extension is not as easily explained, but could possibly be caused by increased activation of mechanoreceptors in facet joints. PMID- 11013502 TI - Functional restoration versus outpatient physical training in chronic low back pain: a randomized comparative study. AB - STUDY DESIGN: A randomized parallel-group comparative trial with a 1-year follow up period was performed. OBJECTIVE: To compare the effect of a comprehensive functional restoration program involving intensive physical training, ergonomic training, and behavioral support (39 hours per week for 3 weeks) with the effect of outpatient intensive physical training (1.5 hours three times per week for 8 weeks). SUMMARY OF BACKGROUND DATA: Nonrandomized studies conducted in the United States favor functional restoration for patients with chronic low back pain. Two previously reported randomized studies from the authors' Back Center in Copenhagen concur with this recommendation, although the positive effects in one of the studies had faded out after 2 years. Randomized functional restoration studies in Canada and Finland have failed to demonstrate any substantive effect. METHODS: Initially, 138 patients with chronic low back pain were included in the current study. They then were randomized to either functional restoration (n = 64) or outpatient intensive physical training (n = 74). Of the initial 138 patients, 11 never started (5 and 6, respectively); 21 dropped out during treatment (8 and 13); and 7 of the graduates did not take part in the 1-year follow-up evaluation (3 and 4). The conclusions were drawn from the 99 patients (48 and 51, respectively) who graduated and participated in a 1-year follow-up evaluation. The median age of the patients was 42 years (range, 21-55 years) The female-to-male ratio was 68 to 31, and the median sick leave days during the preceding 3 years was 180 (range, 0-1080 days). The average back pain was rated 5.5 on a scale of 0 (no pain) to 10 (maximal pain). For these variables, there were no important differences between the groups. However, the functional restoration group tended to be more capable of work at baseline (58% vs 42%; P = 0.09). RESULTS: At the 1-year follow-up evaluation, overall assessment favored functional restoration. Otherwise, no significant differences were observed regarding work capability, sick leave for those at work, health care contacts,back pain, leg pain, or self-reported activities of daily living. CONCLUSIONS: Only in terms of overall assessment, the functional restoration program was superior to a comparatively short time-consuming outpatient physical training program. DISCUSSION: It may be that lower economic benefits during sick leave in the United States lead to favorable results from functional restoration programs, whereas greater benefits in Canada, Finland, and Denmark result in different conclusions. Finally, it may be that the difference in results across studies points simply to whether the studies were randomized. PMID- 11013503 TI - Nonsteroidal anti-inflammatory drugs for low back pain: a systematic review within the framework of the Cochrane Collaboration Back Review Group. AB - STUDY DESIGN: A systematic review of randomized and double-blind controlled trials was performed. SUMMARY OF BACKGROUND DATA: Nonsteroidal anti-inflammatory drugs are the most frequently prescribed medications worldwide and are widely used for patients with low back pain. OBJECTIVES: To assess the effects of nonsteroidal anti-inflammatory drugs in the treatment of nonspecific low back pain with or without radiation, and to assess which type of nonsteroidal anti inflammatory drug is most effective. METHODS: For this study, the Cochrane Controlled Trials Register, Medline and Embase, and reference lists of articles were searched. Two reviewers blinded with respect to authors, institution, and journal independently extracted data and assessed the methodologic quality of the studies. If data were considered clinically homogeneous, a meta-analysis was performed. If data were considered clinically heterogeneous, a qualitative analysis was performed using a rating system with four levels of evidence: strong, moderate, limited, and no evidence. RESULTS: This review involved 51 trials and 6057 patients. Of these trials, 16 (31%) were of high quality. The pooled relative risk for global improvement after 1 week was 1.24 (95% confidence interval [CI] = 1.10-1.41), and for additional analgesic use was 1.29 (95% CI = 1.05-1.57), indicating a statistically significant but small effect in favor of nonsteroidal anti-inflammatory drugs as compared with a placebo. The results of the qualitative analysis showed that there is conflicting evidence (Level 3) that nonsteroidal anti-inflammatory drugs are more effective than paracetamol for acute low back pain, and that there is moderate evidence (Level 2) that nonsteroidal anti-inflammatory drugs are not more effective than other drugs for acute low back pain. There is strong evidence (Level 1) that various types of nonsteroidal anti-inflammatory drugs are equally effective for acute low back pain. CONCLUSIONS: The evidence from the 51 trials included in this review suggests that nonsteroidal anti-inflammatory drugs are effective for short-term symptomatic relief in patients with acute low back pain. Furthermore, there does not seem to be a specific type of nonsteroidal anti-inflammatory drug that is clearly more effective than others. Sufficient evidence on chronic low back pain still is lacking. PMID- 11013504 TI - Primary lumbosacral stability after open posterior and endoscopic anterior fusion with interbody implants: a roentgen stereophotogrammetric analysis. AB - STUDY DESIGN: After posterior stabilization of the spondylolytic lumbosacral level, mobility of the fused vertebrae could be studied before and after an additional anterior endoscopic interbody fusion using roentgen stereophotogrammetric analysis. OBJECTIVE: To determine the in vivo primary lumbosacral stability of additional anterior interbody fusion after transpedicular screw fixation. SUMMARY OF BACKGROUND DATA: In vitro studies indicate a significant decrease in segmental motion after pedicle screw fixation and additional anterior fusion. Roentgen stereophotogrammetric studies demonstrate the adequacy of transpedicular lumbar instrumentation in posterolateral fusions. There are no studies examining the effect of additional anterior interbody fusion after posterior instrumentation in vivo. METHODS: In this study, 15 patients with low-grade spondylolisthesis at L5-S1 underwent a two stage open posterior and endoscopic anterior lumbar fusion using carbon fiber (Brantigan I/F) cages. At surgery, tantalum markers were implanted into the fifth lumbar (L5) and the first sacral (S1) vertebra. All the patients were examined by roentgen stereophotogrammetric analysis after the first and second surgical procedures. RESULTS: After implantation of the posterior pedicle system only, the mean intervertebral mobility determined by roentgen stereophotogrammetric analysis was 0.23 mm in the transverse (x), 0.54 mm in the vertical (y), and 1.2 mm in the sagittal (z) axes. After additional anterior endoscopic fusion with carbon cages, the remaining translation between the fused segment L5/S1 decreased to 0.17 mm in the x, 0.16 mm in the y, and 0.44 mm in the z axes. CONCLUSION: Anterior endoscopic lumbosacral fusion significantly increases the primary stability of the posterior fusion with a pedicle system in two axes of motion. PMID- 11013505 TI - Spinal reflex attenuation associated with spinal manipulation. AB - STUDY DESIGN: This study evaluated the effect of lumbosacral spinal manipulation with thrust and spinal mobilization without thrust on the excitability of the alpha motoneuronal pool in human subjects without low back pain. OBJECTIVES: To investigate the effect of high velocity, low amplitude thrust, or mobilization without thrust on the excitability of the alpha motoneuron pool, and to elucidate potential mechanisms in which manual procedures may affect back muscle activity. SUMMARY OF BACKGROUND DATA: The physiologic mechanisms of spinal manipulation are largely unknown. It has been proposed that spinal manipulation may reduce back muscle electromyographic activity in patients with low back pain. Although positive outcomes of spinal manipulation intervention for low back pain have been reported in clinical trials, the mechanisms involved in the amelioration of symptoms are unknown. METHODS: In this study, 17 nonpatient human subjects were used to investigate the effect of spinal manipulation and mobilization on the amplitude of the tibial nerve Hoffmann reflex recorded from the gastrocnemius muscle. Reflexes were recorded before and after manual spinal procedures. RESULTS: Both spinal manipulation with thrust and mobilization without thrust significantly attenuated alpha motoneuronal activity, as measured by the amplitude of the gastrocnemius Hoffmann reflex. This suppression of motoneuronal activity was significant (P < 0.05) but transient, with a return to baseline values exhibited 30 seconds after intervention. CONCLUSIONS: Both spinal manipulation with thrust and mobilization without thrust procedures produce a profound but transient attenuation of alpha motoneuronal excitability. These findings substantiate the theory that manual spinal therapy procedures may lead to short-term inhibitory effects on the human motor system. PMID- 11013506 TI - The effect of neuromuscular blockade on pedicle screw stimulation thresholds. AB - STUDY DESIGN: Nerve root stimulation thresholds were studied relative to the level of neuromuscular blockade in patients undergoing lumbar decompression surgery. OBJECTIVES: To determine what levels of intraoperative neuromuscular blockade can be used during pedicle screw stimulation. BACKGROUND DATA: Previous studies of intraoperative pedicle screw stimulation thresholds have failed to determine the effect of neuromuscular blockade on the stimulation threshold. METHODS: Twenty-one roots in 10 patients undergoing lumbar decompression surgery were studied at different levels of neuromuscular blockade. Ninety-five nerve root thresholds were determined relative to level of blockade. RESULTS: Neuromuscular blockade below 80% provides nerve root thresholds similar to thresholds without blockade. CONCLUSIONS: Neuromuscular blockade should be less than 80% when using pedicle screw electrical stimulation testing. PMID- 11013508 TI - Follow-up evaluation of resected lumbar vertebral chordoma over 11 years: a case report. AB - STUDY DESIGN: A case report with an 11-year follow-up assessment after resection and reconstruction for lumbar chordoma is given. The literature relevant to this topic is reviewed. OBJECTIVES: To report the long-term outcome in a case of lumbar chordoma, to review the literature on vertebral chordoma, and to outline the rationale for surgical resection in such cases. SUMMARY OF BACKGROUND DATA: Chordoma is a malignant bone tumor that grows slowly, often recurs locally, and metastasizes late. Although different treatment approaches exist, including radiation and surgery, the only curative treatment is early and complete surgical excision of the tumor. Immediate spinal stability must be achieved with appropriate replacement or bone graft with rigid fixation. METHODS: The 11-year follow-up evaluation of a 42-year-old woman with L3 and L4 vertebral body chordoma treated with complete removal, femoral shaft allograft replacement, fusion, and rigid metal fixation is reported. The patient was observed with serial physical examinations, radiographs, and laboratory studies over 11 years. RESULTS: At this writing, 11 years after the resection of the L3 and L4 chordoma, the patient is asymptomatic without evidence of recurrence or metastasis. CONCLUSIONS: As reported, vertebral chordomas are not curable, but the authors' experience contradicts this. The surgeon should aim at a wide, or at least a marginal, excision followed by a stable reconstruction. PMID- 11013507 TI - Respiratory failure in postpneumonectomy syndrome complicated by thoracic lordoscoliosis: treatment with prosthetic implants, partial vertebrectomies, and spinal fusion. AB - STUDY DESIGN: This study investigated the case of a 17-year-old girl with postpneumonectomy syndrome, complicated by a thoracic lordoscoliosis, who was successfully treated with prosthetic implants, partial vertebrectomies, and anteroposterior spinal fusion. OBJECTIVE: To report a unique case and describe the authors' method of treatment. SUMMARY OF BACKGROUND DATA: Postpneumonectomy syndrome is an uncommon complication of pneumonectomy. Many case reports describe successful treatment with insertion of prosthetic implants into the empty hemithorax to shift the mediastinum to its original position. Thoracic lordoscoliosis reportedly has contributed to pulmonary compromise, but no cases have shown its occurrence in the setting of postpneumonectomy syndrome. METHODS: The patient was observed at the National Children's Hospital in Tokyo, referred to Children's Hospital in Los Angeles, California for surgical correction, and followed in Tokyo for the next year. RESULTS: Two prosthetic implants with an injection port for further expansion were positioned in the right hemithorax to restore the mediastinum to its normal position. Anterior discectomies, partial vertebrectomies, and fusion of T5-T10 was performed concurrently. Then 5 days later, posterior spinal fusion of T1-T12 with instrumentation and bone graft were performed to correct the thoracic lordoscoliosis and increase the chest cavity space. At 1 month after the surgery, the patient was extubated after being ventilator dependent for 5 months. At the time of operation, the girl was ventilator dependent and nonambulatory, but 1 year later could participate in all activities of daily living without any oxygen supplementation. CONCLUSIONS: Postpneumonectomy syndrome can be treated successfully with prosthetic implants to restore the normal position of the mediastinum. Thoracic lordoscoliosis can complicate the syndrome and may be corrected to help restore normal pulmonary function. PMID- 11013509 TI - Contralateral spondylolysis and fracture of the lumbar pedicle in an elite female gymnast: a case report. AB - STUDY DESIGN: The case of an elite female gymnast whose pathology started in her 12th year and whose evolution has been exceptional is reported. OBJECTIVE: To present a fracture of the right lumbar pedicle showing complete spontaneous consolidation despite gymnastic practice 15 hours a week. SUMMARY OF BACKGROUND DATA: Lumbar pain, which has an incidence of approximately 75% among young athletes, often results from diseases of the posterior arch of vertebrae in gymnasts, including spondylolysis. The association between unilateral spondylolysis and fracture of the contralateral lumbar pedicle in young athletes is poorly described. METHODS: An elite young female gymnast underwent clinical examination and lumbar radiographs (as systematically required by the French Federation for high-level gymnasts) from 1994 to 1997 to join a sports program in gymnastics. RESULTS: Clinical examination and lumbar radiographs systematically required of an asymptomatic female gymnast allowed the condensation of the right pedicle to be observed before lysis of the left isthmus of L5 in 1994, unilateral lysis of the left isthmus of L5 in 1995, a right pedicular fracture of L5 in 1996, and healing of the pedicular fracture in 1997. CONCLUSION: Inconsistency between radiographs and clinical observations can be noted, and spontaneous consolidation of pedicular fractures can occur despite the practice of the gymnastics 15 hours a week. PMID- 11013510 TI - Spine update: antimicrobial prophylaxis in spine surgery: basic principles and recent advances. PMID- 11013511 TI - Early intervention for back-injured nurses at a large hospital. PMID- 11013512 TI - Luque trolley and convex epiphysiodesis in the management of infantile and juvenile scoliosis. PMID- 11013513 TI - The measurement of static pelvic skeletal asymmetry. PMID- 11013514 TI - Revised Oswestry Disability questionnaire. PMID- 11013515 TI - Cause of idiopathic scoliosis. PMID- 11013516 TI - A computer simulation for the entry-level RN: enhancing clinical decision making. AB - Staff development educators struggle to prepare today's entry-level RN for effective decision making in a new healthcare climate that demands both skill and expertise with patients with high acuity levels. The clinical decision making simulator is an innovative approach to teaching and learning decision-making skills. The authors discuss the development and use of a computer simulation that provides repeated opportunities for clinical decision making for the newly licensed nurse without jeopardizing patient safety. PMID- 11013517 TI - Development of a discharge planning mentorship program. AB - Discharge planning coordinators at a tertiary medical center developed a Discharge Planning Mentorship Program. The group established the educational program to support the autonomy of the staff primary nurse in the discharge planning component of professional practice. To establish the program, the discharge planning coordinators used the underlying principle of "CollaMentoach." The term is an acronym combining the three core concepts of collaboration, mentoring, and coaching. Program planning and marketing, participant selection, curriculum development, and curriculum evaluation are included to guide staff development specialists and nurse managers in the formulation of a program to enhance staff nurse discharge planning skills. PMID- 11013518 TI - Providing mandatory safety information to hospital employees: finding a way that is fast, fun, and effective. AB - An interdisciplinary team of medical center employees transformed a 2-hour mandatory safety class into a 15-minute interactive marathon. The marathon, coupled with a self-learning packet and posttest, not only measured cognitive knowledge but included a component of competence demonstration. The information included 10 specific safety topics pertinent to all medical center employees including those in 12 physician satellite offices. This educational format has proven successful in providing safety information that is fast, fun, and effective. PMID- 11013519 TI - Facilitating critical thinking. AB - Supporting staff to think effectively is essential to improve clinical systems, decrease errors and sentinel events, and engage staff involvement to refine patient care systems in readiness for new care-delivery models that truly reflect the valued role of the RN. The authors explore practical methods, based on current research and national consulting experience, to facilitate the development of mature critical thinking skills. Assessment tools, a sample agenda for formal presentations, and teaching strategies using behavioral examples that make the important and necessary link of theory to reality are discussed in the form of a critical thinking test as well as a conceptual model for application in problem solving. PMID- 11013520 TI - Integrating technology and traditional teaching methods to stimulate different cognitive styles in a critical care course. AB - This article describes a method of instruction designed to stimulate many different cognitive styles in an RN critical care course. The class met on Friday evenings after many of the participants had just completed their workweek. Developing a stimulating classroom experience was imperative to counter the effects of fatigue felt by many of the nurses. A combination of technology and traditional teaching methods was used to facilitate as many different cognitive styles as possible in a single class period. The PowerPoint computer program was used to provide vivid visual representation of the class content as it was presented through a combination of lecture, concept mapping, and case study. PMID- 11013521 TI - A CQI approach to evaluating continuing education: processes and outcomes. AB - Traditional methods of evaluating continuing nursing education have focused on end-of-session feedback and/or testing related to the learning experience. A continuous quality improvement (CQI) framework and associated statistical process control tools provide an alternative approach to assessing both processes and outcomes of continuing nursing education. One implementation example demonstrates the value of this approach. The CQI model offers an exciting option to continuing education/staff development educators looking for creative and beneficial ways to evaluate educational activities. PMID- 11013522 TI - Human responses to pain: a global perspective. PMID- 11013523 TI - Pain as a mutual experience for patients, nurses and families: unique international perspectives from the Peoples' Republic of China, the United States, Malawi and Spain. AB - This article shares findings from a descriptive research study that explored pain recognition, pain intervention and perceived pain relief in laboring women, children and older adult patients from four very different countries. Researchers included co-investigators from each site and a team of more than 20 nurses from China, the United States, Malawi and Spain. This international, multi-site study investigated specific aspects of the phenomena of pain which are known to be of interest to nurses internationally. Critical issues related to feasibility of this study depended, in large part, on the ability of principle investigators to have access to a sample comprised of sub-samples of nationally different subjects that would represent the population of interest. Access to the four study countries was made possible through professional relationships previously developed by either one of the principal or co-investigators. PMID- 11013524 TI - Pain as a mutual experience for patients, nurses and families: a perspective from Shanghai, China. AB - The author of this article visited China for the purpose of helping the faculty of the School of Nursing learn research skills by participating in research--a kind of learn by doing. Both investigators had conducted quantitative studies in the US--one with children and one with adults--that were adapted for use in China. Seeking to also include a qualitative study, the investigators explored several possible research areas. Because pain is a universal phenomena, it was chosen as the subject for the study using the qualitative methodology reported in the first part of this article. PMID- 11013526 TI - Pain as a mutual experience for patients, nurses and families: a perspective from Lilongwe, Malawi. AB - The data for this study were collected in the Republic of Malawi (formerly Nyasaland), a small country in South Central Africa. Data were collected by registered nurses who were senior students in the first class of registered nurse midwives enrolled in a baccalaureate completion program who had completed a research course taught by the CUNY investigator. Fourteen women participated in the study; all were accompanied by a female relative or friend. Because of cultural norms and taboos, paternal attendance in labor settings rarely occurs and women in labor receive support from mothers, aunts and mothers-in-law. In summary, accurate estimation of patient pain among nurses and families occurred less than 25% of the time. PMID- 11013525 TI - Pain as a mutual experience for patients, nurses and families: a perspective from New York. AB - New York was unique among the cities included in this study. The most striking finding was that the self-described ethnic background of both nurses and patients was so varied. Many patients and nurses were born elsewhere and were nonnative speakers. The results cannot be described as cross-countries when so many nations are represented. Multinational is, perhaps, a more accurate description, but even that seems inadequate to describe a population as diverse as was this one. PMID- 11013527 TI - Pain as a mutual experience for patients, nurses and families: a perspective from Valencia, Spain. AB - The Spanish data collection was consistent with the overall study design and drew from three groups: laboring women, children and elderly patients. The Valencia data was amongst the most detailed, specific and complete in this international study. This is most likely due to the experienced nature of the Spanish research team. The study results revealed more commonalities than differences in all age groups with regard to pain identification and pain alleviation. Across age groups, pain was identified by study participants through observation and listening. PMID- 11013528 TI - Pain as a mutual experience for patients, nurses and families: international and theoretical perspectives from the four countries. AB - In Level III, country specific themes, outlined as quantitative and qualitative findings in Level II, were subjected to extended analyses using the same methods, from an international perspective. Aggregated themes, examined for patients, nurses and families, across all four countries are presented in that order. PMID- 11013529 TI - Controversies in the treatment and management of Lyme disease. AB - Lyme disease is an infection caused by the bite of the deer tick. It is the most common vector-transmitted disease in the United States. Past treatment practices included the administration of oral and/or i.v. antibiotics. Today the question is "to treat or not to treat." The etiology, signs and symptoms, and current treatments and management of Lyme disease and its associated somatic complaints are discussed. PMID- 11013530 TI - Certification and credentialing to define competency-based practice. AB - One of the challenges that healthcare organizations face today is how to effectively establish, document, and evaluate competency in intravenous practice. The process used to define practice parameters based on national certification and credentialing versus organizational policies is described. Key attributes, the role of mentoring, and adverse outcomes are addressed. PMID- 11013531 TI - Efficacy of ultrasonography in peripheral venous cannulation. AB - A retrospective study of 431 patients who had peripherally inserted midclavicular or central catheters placed during a consecutive 13-month period using the conventional landmark method for placement was compared with a second group of 326 patients, who during a 12-month period had such catheters placed using ultrasonography. The data demonstrate a 42% decrease in the number of needle penetrations needed to successfully cannulate veins when ultrasound was used during placement. There is a 26% greater chance of successful cannulation of the vein on the first attempt with ultrasound-guided placements than with those using the traditional landmark method. PMID- 11013532 TI - Tracking catheters: the care continuum. AB - Care coordination and development of a plan to track vascular access devices (VADs) is presented. The article discusses how to establish criteria so that central venous catheters can be monitored. Specific areas of focus for monitoring outcomes are reviewed, as is the process of obtaining invaluable information. This information can be used to support policy and procedure changes and clinical practice changes to provide quality outcomes. PMID- 11013533 TI - Cellular effects of cancer chemotherapy administration. AB - Cancer chemotherapy drugs (CDs) exert a cytotoxic effect by altering one or more aspects of the growth kinetics of cancer and malignant cells. The pattern and extent of cytotoxicity determines in part the scientific rationale of various CD regimens and accounts for many of the common side effects of treatment. This article provides an overview of the behavior of cancer at the cellular level and the impact exerted by CDs on normal and malignant cell growth. Examples are provided of how these scientific concepts affect treatment outcomes and clinical decisions. The nurse's role in monitoring and managing the related side effects of treatment also is discussed. PMID- 11013535 TI - Fitness for practice. PMID- 11013534 TI - Ethics and total parenteral nutrition: issues for intravenous nurse professionals. AB - Healthcare ethics is a hot topic these days. Decisions to withhold or withdraw various forms of medical therapy are daily events in most hospitals and long-term care facilities. Intravenous nurse professionals do not need to be bioethicists; however, they do need to be able to identify problems quickly and know how to address them. Some of the ethical issues that could be encountered by i.v. nurse professionals involved in the provision of total parenteral nutritional support in various clinical settings are examined and explored. PMID- 11013536 TI - Do new roles contribute to job satisfaction and retention of staff in nursing and professions allied to medicine? AB - BACKGROUND: Studies have suggested that job dissatisfaction is a major factor influencing nurses' and occupational therapists' intention to leave their profession. It has also been related to turnover of qualified nurses. However, literature relating to these factors among nurses and professions allied to medicine in innovative roles is scarce. AIMS: This paper considers the views of 452 nurses and 162 professionals allied to medicine (PAMs) in innovative roles, on job satisfaction, career development, intention to leave the profession and factors seen as hindering and enhancing effective working. METHODS: A self completion questionnaire was developed as part of a larger study exploring new roles in practice (The ENRiP Study). FINDINGS: Overall there was a high level of job satisfaction in both groups (nurses and PAMs). Job satisfaction was significantly related to feeling integrated within the post-holder's own professional group and with immediate colleagues, feeling that the role had improved their career prospects, feeling adequately prepared and trained for the role, and working to protocol. Sixty-eight percent (n = 415) of respondents felt the role had enhanced their career prospects but over a quarter of respondents (n = 163; 27%) said they would leave their profession if they could. Low job satisfaction was significantly related to intention to leave the profession. CONCLUSIONS: The vast majority of post-holders in innovative roles felt that the role provided them with a sense of job satisfaction. However, it is essential that the post-holders feel adequately prepared to carry out the role and that the boundaries of their practice are well defined. Career progression and professional integration both being associated with job satisfaction. PMID- 11013537 TI - The nurse executive: challenges for the 21st century. AB - AIM: The aim of this paper is to examine the challenges facing the nurse executive in the 21st century by questioning the traditional attributions of leadership to the nurse executive role. BACKGROUND: Historically, the leadership role in nursing has been assumed by the nurse executive. The predominantly female character of nursing, however, has ensured that demonstrations of leadership amongst nurses have been infrequent and compatible with prevailing male-defined ideologies. Examples of this include career restructuring and educational reforms in Australia. FINDINGS: This paper found that the apparent lack of leadership in nursing was able to be traced back to early management theories which categorized leadership as a function of management. CONCLUSIONS: If nurses are to assume leadership positions in the health care system of the 21st century, nurse leaders will have to let go of traditional managerial practices and behaviours. In the emerging health care system of the new century, nurse executive practices will focus on achieving change rather than predictability in organizational outcomes. PMID- 11013538 TI - Can information technology help ward sisters become ward managers? AB - AIM: This paper describes an evaluation methodology that focuses on management decisions at the ward level in a hospital. BACKGROUND: Successive governments in the 1990s have provided considerable funds to introduce decision support systems for ward management, but few have been evaluated rigorously. This paper reviews the approaches to systems' evaluation that have been developed. METHODS: Measuring instruments were designed to define and measure management decisions and those organizational factors which affect decision making. The resulting difficulty index was validated in a before-and-after study of ward decision making. FINDINGS: Changes in decision making, after the implementation of a decision support system, were found to relate to the identified organizational factors. CONCLUSION: The evaluation methodology described in this paper showed that the potential benefits of a computerized management system would not be realized unless managers, doctors and nurses carried out an organizational review before implementation and then took joint ownership of the system. PMID- 11013539 TI - Visual art dialogues with elderly persons: effects on perceived life situation. AB - AIM: The investigation was aimed towards constructing a visual art programme for communication with elderly. METHODS: Pictures of works of art were used in a controlled intervention study. Dialogues were performed with elderly persons (age 82.6 years) at a senior's apartment building. The Wheel Questionnaire parameters structure, motivation, and emotional investment were analysed using ANOVA (mixed model). FINDINGS: Significant improvement was found in the visual art group (n = 20) compared with a matched control group (n = 20) over the studied period of time. Communication directions were different in the intervention group compared with the control group. In the intervention group there was an inexhaustible source of topics to be discussed that originated from pictures of works of art. In the control group the dialogues dealt with daily events in the elderly persons' lives. During the final phase of the intervention period it was difficult to find topics of conversation in the control group compared with the intervention group. CONCLUSIONS: The visual art programme is an example of how the language of works of art could be used for nursing management. The findings show a new way to care for elderly persons that builds upon elderly persons' knowledge and personal experience. PMID- 11013540 TI - Direct care staff's need for support in their perceived work role in day activities units. AB - AIMS: This study sets out to investigate direct care staff's views of their need for support, supervision, and training in their practice in day care settings when supporting daily occupations among developmentally disabled persons. BACKGROUND: The first line staff are considered as having a prominent role in the successful delivery of service. METHODS: Three municipalities, one urban and two rural areas in southern Sweden, were chosen for the study. The studied population n = 81 consisted of 94.1% of all staff employed in day activities units supporting the clients' daily occupations or community-integrated, sheltered work employment. The data were collected by means of a questionnaire. FINDINGS: This material identified the staff's perceived work role, and their needs for support, supervision, and training in the areas of communication, environment adaptation, individual activation and training methods, with regard to learning disabilities and special needs. CONCLUSIONS: Care managers should focus upon preparation of staff support programmes to improve the quality and efficiency in this area of care. PMID- 11013541 TI - Is access to a standardized neonatal intensive care possible? AB - AIMS: This paper aims to determine the factors which impact on the issue of availability and access to a standard quality of neonatal intensive care provision in order to clarify strategies for change. BACKGROUND: New targets set within the NHS reforms have highlighted quality and fair access to services. Current frameworks for the provision of neonatal services may not be in place to support a standardized quality service provision. METHODS: The paper examines the historical and political basis for current models of service, giving a grounding for recommendations for change in an attempt to achieve a more standardized, equitable level of care. FINDINGS: Changes within health care policy in the late 1980s and 1990s have led to confusion within the service and a lack of the definition needed to provide universally accepted criteria on which to base a regional service provision. CONCLUSION: The organization of services will need to be reviewed to include the introduction of national and regional data collection and analysis, and national definitions standardizing service criteria to be used to support variations in regional models of care. In conjunction with these recommendations independent auditing processes must exist to allow for accreditation of services. PMID- 11013542 TI - Communication: the royal pathway to patient-centered medicine. PMID- 11013543 TI - The enduring and evolving nature of the patient-physician relationship. AB - Just as the molecular and chemistry oriented sciences were adopted as the 20th century medical paradigm, incorporation of the patient's perspective into a relationship-centered medical paradigm has been suggested as appropriate for the 21st century. It is the medical dialogue that provides the fundamental vehicle through which the paradigmatic battle of perspectives is waged and the therapeutic relationship is defined. In many regards, the primary challenge to the field is the development of operationally defined and measurable indicators of medical communication that will provide a valid representation of the conceptual models of the therapeutic relationship. The purpose of this essay is to explore the implications of a relationship-centered medical paradigm on the nature of the patient-physician relationship and its expression in the communication of routine medical practice. An organizing framework for distinguishing commonly measured communication elements into conceptually distinct components is suggested. Application of this framework is illustrated through an empirical study of communication in primary care practice. The results of the study demonstrate the usefulness of this approach in linking communication to models of therapeutic relationships. The importance of medical communication is further explored in a summary of studies that establish its association to outcomes and in an overview of future challenges to the field. PMID- 11013544 TI - Bridging the gap. The separate worlds of evidence-based medicine and patient centered medicine. AB - Modern medical care is influenced by two paradigms: 'evidence-based medicine' and 'patient-centered medicine'. In the last decade, both paradigms rapidly gained in popularity and are now both supposed to affect the process of clinical decision making during the daily practice of physicians. However, careful analysis shows that they focus on different aspects of medical care and have, in fact, little in common. Evidence-based medicine is a rather young concept that entered the scientific literature in the early 1990s. It has basically a positivistic, biomedical perspective. Its focus is on offering clinicians the best available evidence about the most adequate treatment for their patients, considering medicine merely as a cognitive-rational enterprise. In this approach the uniqueness of patients, their individual needs and preferences, and their emotional status are easily neglected as relevant factors in decision-making. Patient-centered medicine, although not a new phenomenon, has recently attracted renewed attention. It has basically a humanistic, biopsychosocial perspective, combining ethical values on 'the ideal physician', with psychotherapeutic theories on facilitating patients' disclosure of real worries, and negotiation theories on decision making. It puts a strong focus on patient participation in clinical decision making by taking into account the patients' perspective, and tuning medical care to the patients' needs and preferences. However, in this approach the ideological base is better developed than its evidence base. In modern medicine both paradigms are highly relevant, but yet seem to belong to different worlds. The challenge for the near future is to bring these separate worlds together. The aim of this paper is to give an impulse to this integration. Developments within both paradigms can benefit from interchanging ideas and principles from which eventually medical care will benefit. In this process a key role is foreseen for communication and communication research. PMID- 11013545 TI - Evidence-based guidelines for teaching patient-centered interviewing. AB - In a rare study of effectiveness of an interviewing method, we previously reported a randomized controlled trial demonstrating that training in a step-by step patient-centered interviewing method improved residents' knowledge, attitudes, and skills and had a consistently positive effect on trained residents' patients. For those who wish to use this evidence-based patient centered method as a template for their own teaching, we describe here for the first time our training program--and propose that the training can be adapted for students, physicians, nurse practitioners, physician assistants, and other new learners as well. Training was skills-oriented and experiential, fostered positive attitudes towards patient-centered interviewing, and used a learner centered approach which paid special attention to the teacher-resident relationship and to the resident's self-awareness. Skills training was guided by a newly identified patient-centered interviewing method that described the step by-step use of specific behaviors. PMID- 11013546 TI - Differences in counseling men and women: family planning in Kenya. AB - A comparison of family planning sessions with male and female clients in Kenya found distinct gender differences. Most men came for information, while women wanted to adopt, continue, or change contraceptive methods. Consultations with men and couples were more than twice as long as consultations with women. Men communicated actively (for example, by volunteering extra information, asking questions, and expressing worries) during 66% of their turns to speak, compared with 27% for women. Providers offered men more detailed information than women, asked them fewer questions, issued fewer instructions, and responded more supportively. These communication patterns may be seen as a reflection of Kenyan gender roles and men's and women's different reasons for seeking family planning services. Kenyan providers need to improve the quality of their interactions with women. They also need to anticipate men's outspokenness and understand the male agenda if they are to counsel men effectively. PMID- 11013547 TI - The practice orientations of physicians and patients: the effect of doctor patient congruence on satisfaction. AB - This study investigated the extent to which the individual orientations of physicians and patients and the congruence between them are associated with patient satisfaction. A survey was mailed to 400 physicians and 1020 of their patients. All respondents filled out the Patient-Practitioner Orientation Scale, which measures the roles that doctors and patients believe each should play in the course of their interaction. Patients also rated their satisfaction with their doctors. Among patients, we found that females and those who were younger, more educated, and healthier were significantly more patient-centered. However, none of these variables were significantly related to satisfaction. Among physicians, females were more patient-centered, and years in practice was related to satisfaction and orientation in a non-linear fashion. The congruence data indicated that patients were highly satisfied when their physicians either had a matching orientation or were more patient-centered. However, patients whose doctors were not as patient-centered were significantly less satisfied. PMID- 11013548 TI - Pharmacists' evaluation of key communication skills in practice. AB - This paper reports the results of a major research initiative into the identification of key communication skills in community pharmacist-patient consultations. It is now widely accepted that the quality of practitioner-patient communication is fundamental to effective health care. However, an analysis of the literature pertaining to the communication issues facing health professionals in general and pharmacists in particular emphasised the need for more empirical research, to chart what pharmacists themselves deemed to be the nature and range of skills which contribute to effective communication performance in community pharmacy practice. The main aim of this research investigation was, therefore, to identify what constituted effective communicative performance by community pharmacists. This paper provides full details of the repertoire of skills and sub skills identified as being the core communicative elements of practice. The results of this research will have relevance for health professionals and behavioural scientists, and will also contribute to the assurance of quality within the field of community pharmacy practice. PMID- 11013549 TI - Measuring patient-centredness: a comparison of three observation-based instruments. AB - The aim of this study was to assess the reliability and validity of three different observation-based measures of patient-centredness. The three face-valid instruments were each applied to the same sample of 55 videotaped GP consultations. Associations were explored with consultation 'input' variables (e.g. patient and doctor demographic characteristics, patient health status) and 'process' variables (e.g. consultation length). The three measures demonstrated varying levels of inter-rater reliability. Reliability was proportional to training requirements. Differences in construct validity of the three measures were evident and their concurrent validity was relatively low. Researchers must exercise caution in their choice of measurement method because of differences in how the concept of 'patient-centredness' is operationalized. Greater conceptual specificity and simplification are required for meaningful, reliable measurement. The implications for research, and for assessing the quality of individual doctors' 'interpersonal' care are discussed. PMID- 11013550 TI - Decision-making in nephrology: shared decision making? AB - Shared decision-making is considered an important ideal for physician-patient interaction. The ideal states that health-related values should be discussed together. It raises two questions: (a) for which decisions is the ideal of shared decision-making relevant? (b) Which aspects of treatment should be discussed? The nephrological practice under consideration in this article answers question (a) as follows: decisions about the type of dialysis are shared decisions, while decisions about the moment to start dialysis are medical decisions that should be taken by nephrologists. This situation can be criticized as important health related values play a role in decisions about starting dialysis. Question (b) is answered in the nephrological practice under consideration by discussing at least all important health-related aspects that raise uncertainty about its worth for a patient. This approach to question (b) is morally and practically defensible. PMID- 11013551 TI - Effects of video interaction analysis training on nurse-patient communication in the care of the elderly. AB - This paper describes an empirical evaluation of communication skills training for nurses in elderly care. The training programme was based on Video Interaction Analysis and aimed to improve nurses' communication skills such that they pay attention to patients' physical, social and emotional needs and support self care in elderly people. The effects of the training course were measured in an experimental and control group. They were rated by independent observers, by comparing videotapes of nursing encounters before and after training. Forty nurses participated in 316 videotaped nursing encounters. Multi-level analysis was used to take into account similarity among same nurse encounters. It was found that nurses who followed the training programme, provided the patients with more information about nursing and health topics. They also used more open-ended questions. In addition, they were rated as more involved, warmer and less patronizing. Due to limitations in the study design, it could not be demonstrated that these findings can entirely be ascribed to the training course. Further research, incorporating a randomized controlled design and larger sample sizes, is recommended to determine whether the results can be attributed to this specific type of training. PMID- 11013552 TI - Patients who present physical symptoms in the absence of physical pathology: a challenge to existing models of doctor-patient interaction. AB - Many patients seek physical treatment for physical symptoms in the absence of physical pathology and incur symptomatic interventions that are ineffective, costly and iatrogenic. It is therefore important to understand how decisions to provide physical intervention can arise in consultations in the absence of physical pathology. Existing models of doctor-patient communication are ill suited to understanding these consultations. A series of studies has provided the components of an alternative approach that is based on understanding consultation from the patients' perspective. Specifically, these studies have delineated: sources of patients' perception of their authority over doctors; what patients seek by consulting their doctors; and ways that patients use their authority to influence doctors to provide what they seek. Patients' authority reflects primarily their own sensory and infallible knowledge of symptoms. Their influence derives from descriptions of subjective symptoms and from additional strategies including descriptions of the psychosocial effects of symptoms, catastrophising and requesting treatment. This analysis suggests directions for future research and medical training. PMID- 11013554 TI - Evaluation of communication training programs in nursing care: a review of the literature. AB - An important aspect of nursing care is communication with patients. Nurses' major communication tasks are not only to inform the patient about his/her disease and treatment, but also to create a therapeutically effective relationship by assessing patients' concerns, showing understanding, empathy, and providing comfort and support. In this review, 14 studies, which focus on the evaluation of the effects of communication training programs for nurses, have been evaluated. The selected studies were screened on several independent, process and outcome variables as described by Francke et al. [8]. In this way not only is the training program taken into account as a variable which may be responsible for nurses' behavioural change and for changes in patient outcomes, but also a range of other variables which can give more nuanced explanations for a training program's degree of effectiveness. On the whole, the studies reviewed showed limited or no effects on nurses' skills, on nurses' behavioural changes in practice, and on patient outcomes. Finally, the majority of the studies had a weak design. The use of experimental research designs should be pursued in future studies in order to eliminate the influence of confounding variables. PMID- 11013553 TI - Doctor-patient communication in different European health care systems: relevance and performance from the patients' perspective. AB - Our aim is to investigate differences between European health care systems in the importance attached by patients to different aspects of doctor-patient communication and the GPs' performance of these aspects, both being from the patients' perspective. 3658 patients of 190 GPs in six European countries (Netherlands, Spain, United Kingdom, Belgium, Germany, Switzerland) completed pre and post-visit questionnaires about relevance and performance of doctor-patient communication. Data were analyzed by variance analysis and by multilevel analysis. In the non-gatekeeping countries, patients considered both biomedical and psychosocial communication aspects to be more important than the patients in the gatekeeping countries. Similarly, in the patients' perception, the non gatekeeping GPs dealt with these aspects more often. Patient characteristics (gender, age, education, psychosocial problems, bad health, depressive feelings, GPs' assessment of psychosocial background) showed many relationships. Of the GP characteristics, only the GPs' psychosocial diagnosis was associated with patient reported psychosocial relevance and performance. Talking about biomedical issues was more important for the patients than talking about psychosocial issues, unless the patients presented psychosocial problems to the GP. Discrepancies between relevance and performance were apparent, especially with respect to biomedical aspects. The implications for health policy and for general practitioners are discussed. PMID- 11013556 TI - [Nursing advise concerning nutritional intake: selecting food is more than just feeding]. PMID- 11013555 TI - [Assisting diabetics: nutrition counseling and health advise]. PMID- 11013557 TI - [Enteral feeding: hygiene as the primary rule]. PMID- 11013558 TI - [Health aspects of professional clothing. 1. Textile eczema is not a rarity]. PMID- 11013559 TI - [Wellness and fitness in old age homes: enhancing independence and quality of life for seniors]. PMID- 11013560 TI - [Staying healthy in everyday working life: stress factors in the nursing profession and how to avoid them]. PMID- 11013561 TI - [Demands on the nursing project manager: motivation through emotional intelligence]. PMID- 11013562 TI - [Problem-oriented learning: activating case studies and increasing learning results]. PMID- 11013563 TI - [Developing emotional competence: improving dealing with patients' emotions]. PMID- 11013564 TI - Are children's nurses fit for the next millennium? PMID- 11013565 TI - Child growth: "Hall" turned upside down. PMID- 11013566 TI - Comparison of flow rates of holes versus cross-cut teats for bottle-fed babies. AB - Under laboratory conditions the flow rates of two types of teat were compared. The openings of Group A teats were in the form of a cross-cut measuring 1.5 mm in radius plus a small central hole. Group B teats were pierced with a single hole of 0.5 mm in diameter. Five teats from each group were tested under atmospheric pressure conditions simulating the effect of a baby's sucking. The results of this trial confirms the hypothesis that the style of teat affects the flow rate and that faster flow rates can be obtained with a cross-cut teat rather than a single-hole teat. Babies using cross-cut teats may also learn to control flow rates by changing their sucking style. PMID- 11013567 TI - Fetal echocardiography and congenital heart disease. AB - Fetal echocardiography has proved a useful tool for prenatal detection of cardiac lesions and the diagnosis--and, in some cases, the treatment--of fetal arrhythmias. It is particularly indicated for mothers from high risk groups. Management of diagnosed heart disease leads either to termination of pregnancy or to optimal postnatal care for the baby and the mother. PMID- 11013568 TI - Nurse practitioner at a GP surgery. PMID- 11013569 TI - Advising on a thermometer for family use. AB - Families will find it useful to have a clinical thermometer, but parents should be taught that children's illnesses can present in a range of different ways and that a young baby can be ill despite having a normal temperature. For people who find mercury thermometers difficult to use and read, alternatives are digital, strip or ear thermometers. Digital thermometers are safe, accurate and very easy to read. Colour-strip thermometers are placed on the skin and change colour at certain temperatures. As well as forehead strips there is a newer kind which can be used under the tongue. Ear thermometers are increasingly popular for young children, being safe, easy to read and accurate. PMID- 11013570 TI - Enuresis: sharing new research and practice. AB - We now understand more about the causes and treatment but must work to overcome the stigma associated with enuresis. Training for professionals needs to be standardised. Terminology also needs to be standardised. European research suggests that the quality of the relationship between professional and child can affect the outcome of treatment. A range of treatments is possible but the first step is a clear assessment of the problem and cause. It is important to take account of the family setting. Never assume incontinence is inevitable for children with special needs and that nothing can be done. Much can be achieved with appropriate training programmes. PMID- 11013571 TI - Nappy rash: let's give mothers more help. AB - Nappy rash affects most babies at some stage. It can occur at any time during the nappy-wearing years, but reaches a peak between 7 and 12 months. Nappy rash causes discomfort to babies and distress to mothers, who often feel guilty and ill-prepared for this problem. Keeping the nappy area dry is the most important factor in prevention and treatment. Recommend frequent changes of nappy and leaving the area uncovered when possible. Thrush is present in many cases of nappy rash. Keeping the area dry deprives thrush of the opportunity to thrive. PMID- 11013572 TI - [Old age homes: in the process of reform]. PMID- 11013573 TI - [Violence and mistreatment]. PMID- 11013574 TI - [Patient care procedures in homes for the aged]. PMID- 11013576 TI - [Nursing support in an old age home]. PMID- 11013575 TI - [Nursing counseling in incontinence: a function to be developed]. PMID- 11013577 TI - [Palliative care, delay in France]. PMID- 11013578 TI - Cultural values in health care: questions and answers. PMID- 11013579 TI - Capital investment financing. PMID- 11013580 TI - Cultural values in health care: a personal view. PMID- 11013581 TI - Strategies for nurse managers in the 21st century. PMID- 11013582 TI - Promoting collaborative practice with culturally diverse populations. AB - Practicing nurses in multiple health care settings must be prepared to manage a diverse, multicultural patient population and work collaboratively with co workers representing different cultural backgrounds. To prepare professional nurses to practice effectively with our increasingly diverse population, frontline managers and nursing service administrators must act jointly to increase their knowledge of cultural diversity, share that knowledge, and combine forces to promote cultural awareness and sensitivity in practice settings. PMID- 11013583 TI - Effective use of interpreters in health care: guidelines for nurse managers and clinicians. AB - Because of the rapid growth of the non-English-speaking population in the United States, nurses are increasingly likely to be called on to provide care for patients whose first language is not English. As a result, nurse managers may need to make interpreters available to health care providers in a variety of settings to facilitate effective communication and provide quality care. The purpose of this article is to present nurse managers and nurse clinicians with information that will enable them to work effectively with interpreters. PMID- 11013584 TI - Leadership style for facilitating the integration of culturally appropriate health care. AB - The dramatic and rapid changes occurring in health care present many challenges and opportunities for nurse leaders as they deal with a culturally diverse society. Leadership style is an important consideration for a health care environment that promotes culturally appropriate care. This article presents a theoretical model that consists of management of change, principle-centered leadership, and transformational leadership, and describes how each of these concepts can assist in creating an environment that fosters culturally appropriate health care. PMID- 11013585 TI - The role of nursing administrators in empowering scholarly productivity among clinicians. AB - The authors discuss strategies nursing administrators can use to promote scholarly productivity among their nursing staff. By incorporating mentoring, promoting collegial teams, involving nurses in the development of goals for scholarly activity, promoting professionalism, and modifying assignments to ensure time for research and writing, nurse executives can significantly enhance scholarly productivity in their professional nursing staff. PMID- 11013586 TI - Providing care to a culturally diverse community: a mobile academic nursing center. AB - Health care institutions are increasingly collaborating with nursing education programs to provide quality and cost-effective primary health care services to the community. Mobile academic nursing centers are emerging as viable alternative sites for these services because they can provide immediate access to health care services and circumvent logistical problems in access to care. Although not a new concept, their development as an integral component of the nursing academic unit is being reexamined in light of health care reform and nursing's role in this reformation. Unlike stationary clinics, these "clinics on wheels" provide multisite access to diverse populations and communities. This article describes how a mobile academic nurse managed center can provide nurses and students with opportunities to develop competencies in community-based experiences. PMID- 11013587 TI - Toward a caring culture in professional nursing. AB - Nursing is a profession founded on the traditional value of caring. Faced with rapid and tumultuous change in the health care system, nurses are finding the environment of practice more stressful and less supportive. The ideal of nursing as a caring community is at risk because of economic and political forces. The authors review the impact of managed care and reaffirm the notion of the caring community in the nursing profession. Specific assessment guidelines and recommendations for the caring community are set forth for practitioners working in a variety of organizational settings. Organizational culture is emphasized as the context for caring practice. PMID- 11013588 TI - Integrating cultural diversity in patient education. AB - Diversity among clients in the American health care system is increasing as the population of the United States changes in composition. Health providers, educators, and supervisors are in unique positions to enhance patient education among members of diverse cultures by incorporating cultural research and health beliefs into patient and staff education. Using a culturally defined framework, health providers can more holistically assess the client and subsequently plan culturally appropriate care. PMID- 11013589 TI - The impact of market-based 'reform' on cultural values in health care. AB - The many issues managed care poses for providers and health networks are crystallized in the moral problems occasioned by its shifting of the financial risks of care from insurer to provider. The issues occasioned by market-based reform include: the problems presented by clashes between public expectations and payer restrictions; the corporatization of health service delivery and the cultural shift from humanitarian endeavor to business enterprise the depersonalization of treatment as time and money constraints stretch resources, and the culture rewards efficient "business-like" behavior the underfunding of care for the poor and uninsured, even as these populations grow the restructuring of care and reengineering of healthcare roles as the emphasis shifts from quality of care to conservation of resources rapid mergers of both health plans and institutional providers with all the inherent turmoil as rules change, services are eliminated, and support services are minimized to save money the unhealthy competition inherent in market-based reform that posits profit taking and market share as the measures of successful performance the undermining of the professional ethic of advocacy the use of incentives that pander to greed and self-interest. The costs of sophisticated technologies and the ongoing care of increasingly fragile patients have pulled many other elements into what previously were considered "privileged" professional interactions. The fact that very few citizens indeed could pay out-of-pocket for the treatment and ongoing care they might need led to social involvement (few people remember that both widespread health insurance and public programs are relatively recent phenomena- only about 30 years old). However, whether in tax dollars or insurance premiums, other people's money is being spent on the patient's care. Clearly, those "other people" never intended to give either the patient or the professional open-ended access to their collective pocketbooks. Just what form their involvement ought to take is being tested as "managed care" attempts to control the costs. What limits are acceptable to providers?: lower profit margins? quality controls? acceptable risk levels? To patients?: restricted choice? restricted mobility? restricted access to high tech? And to the general public?: decreased access to high tech? higher taxes? underserved populations? Abandonment of the sick or poor? Which "techniques" are acceptable, and which are not?: risk-sharing with providers? financial incentives for decision makers? rationing access? imposing behavioral parameters? The issues posed by market-based managed care cannot be adequately addressed merely in terms of social resources, nor will answers be found in subordinating human rights to practical materialism. Negotiating ethical guidelines for the "safe" handling of such problems to the good of individuals and of society requires a revitalization of the "old" values: the old commitment to master craftsmanship and altruism, the old emphasis on patient advocacy and human rights. However, these old values must be applied with the "new" knowledge of lifestyle choices (and thus personal responsibility), likely outcomes (and thus reasonable options), and the limits of success (and thus fair redeployment of health resources). PMID- 11013590 TI - [Ethics and rights of hospital users]. PMID- 11013591 TI - [Taking care of patients, a question of ethics and equipment]. PMID- 11013592 TI - [Between resentment and companionship: ethics]. PMID- 11013593 TI - [Ethics of abstention from or stoppage of therapy during resuscitation]. PMID- 11013594 TI - [Questions of ethics concerning the legislation of July 29, 1994, in relation to bioethics]. PMID- 11013595 TI - [Ethics, public hospitals and administration]. PMID- 11013596 TI - [Hospital management and professional ethics]. PMID- 11013597 TI - [Ethics of AP-HP (Public Assistance--Hospitals of Paris)]. PMID- 11013598 TI - [Pedagogic and ethical relations in the formation of health care teams]. PMID- 11013599 TI - [Function of the nursing team and its ethical responsibility]. PMID- 11013600 TI - [Educational use of the Internet in nursing care]. PMID- 11013602 TI - [An evaluation system for efficient training]. PMID- 11013601 TI - [Analysis of diagnostic reasoning of nursing students]. PMID- 11013603 TI - [Ethics of documentation, ethics of the usage of documentation]. PMID- 11013604 TI - Clinical assessment of the reliability of the examination (CARE). PMID- 11013605 TI - Prediction of risk for patients with unstable angina. PMID- 11013606 TI - [Carcinoma of the stomach. Role of imaging for primary diagnosis and preoperative tumor staging]. AB - The aim of imaging of gastric carcinoma has to be to diagnose the carcinoma as early as possible and to sort out the tumors that are resectable. At the same time imaging of gastric carcinoma should reduce the number of futile laparotomies in patients with advanced, non-resectable tumors to a minimum. Today, endoscopy is the method of choice to diagnose gastric carcinoma. Endosonography is advantageous if small carcinomas have to be judged for depth of tumor infiltration (early gastric carcinoma vs. advanced gastric carcinoma) because high resolution images of the gastric wall are obtained. Staging of large carcinomas that have grown beyond the gastric wall is best performed by hydro-CT because CT scans most accurately delineate infiltrations of surrounding organs/structures and help to plan surgery. Nowadays MRI is not suited for gastric imaging. MRI, however, has the highest potential to fundamentally improve staging of gastric carcinoma if high resolution imaging of the gastric wall is combined with screening for metastases in one examination. PMID- 11013608 TI - [Magnetic resonance imaging of hemorrhagic transformation in ischemic posterior infarction]. AB - PURPOSE: To describe the incidence, time course, and clinical correlates of hemorrhagic transformation (HT) of ischemic stroke in the posterior cerebral artery territory. METHODS: Within 42 months 48 patients with 52 occipital lobe infarctions were examined by T1- and T2-weighted MR imaging. The extent and distribution of secondary hemorrhage were analysed at different intervals after stroke. Volume of ischemic and hemorrhagic infarction was measured planimetrically. RESULTS: HT was observed in 71% of the infarcts between the 5th day and up to 1 year after stroke. HT was most frequently (88%) observed in the 2nd and 3rd month. HT was present in 55% of small infarcts (< 10 cm3), in 88% of medium size (10-50 cm3), and in all large (> 50 cm3) infarcts. In 92% HT presented with petechial bleedings within the cortex (64%) or less frequently (28%) in subcortical structures. The latter types of HT showed no progression and did not increase the clinical deficits. Space-occupying bleedings occurred in only two large defects. CONCLUSIONS: In ischemic posterior infarction, HT can frequently be detected within the first three months after stroke and is predominantly of the petechial type and seems not to be relevant with regard to clinical deficits. PMID- 11013607 TI - [Whole-body MRA on a rolling table platform (AngioSURF)]. AB - PURPOSE: Development of a technique for whole-body MR angiography based on a rolling table platform and integration of a surface coil. MATERIAL AND METHODS: The developed rolling table platform AngioSURF (System for Unlimited Rolling Field-of-view) with integrated surface coil can be mounted on top of the original patient table of a Siemens Symphony System. Data acquisition was performed with a standard body array surface coil. The system was tested on three volunteers and one patient with angiographically documented vascular pathology. Data acquisition was performed with a 3D-FLASH-sequence (TR/TE 2.1/0.7 ms, flip angle: 20 degrees, FOV 40 x 40 cm, 80 partitions, matrix 512 x 420 with zero interpolation). Five data sets were collected in immediate succession during continuous injection of a paramagnetic contrast agent. Time of acquisition per data set was 10 seconds. Table repositioning was performed manually within 3 seconds. Thus the total acquisition time amounted to 72 seconds. RESULTS: No problems with handling occurred in any of the four cases. The excellent image quality enables detailed assessment of the displayed vascular territories. CONCLUSIONS: The rolling table platform with integrated surface coil (AngioSURF) allows diagnostic display of the arterial vascular system from supraaortic vessels to the distal trifurcation arteries in only 72 seconds. PMID- 11013609 TI - [MRI of the prostate after combined radiotherapy (afterloading and percutaneous): histopathologic correlation]. AB - AIM: To identify the MRI changes of the prostate after combined (high-dose rate interstitial with external beam) radiotherapy for, localized prostate cancer and to correlate the findings with histology in order to determine the value of MR imaging in the follow-up of these patients. MATERIAL AND METHODS: Twenty-three patients underwent MR imaging at 1.5 T between 6 and 24 months after completion of combined radiotherapy. The prostate was imaged with axial and coronal T2 weighted sequences and axial T1-weighted sequences before and after intravenous administration of Gd-DTPA. Quadrant or sextant biopsy was performed in all cases and three patients with proven persistence of the tumor underwent salvage prostatectomy. The MRI findings were compared with the biopsy results or the large-area sections. RESULTS: On T2-weighted images the fibrotically changed peripheral zone was hypointense while persistent tumor tissue showed hyperintensity. Solid tumors were depicted when they had a diameter of 1 cm or more. Persistent tumors of the diffuse multifocal type escaped detection. Contrast-enhanced T1-weighted imaging yielded no additional information. The accuracy in detecting persistent tumor was 74%. CONCLUSIONS: Histopathologic changes seen after combined radiotherapy correlate with the findings on T2 weighted MR images. MR imaging cannot replace follow-up by routine biopsy. Its only role is assessing local operability in cases found to have increasing PSA levels during follow-up. Further studies are needed to determine the role of MR imaging in this patient population. PMID- 11013610 TI - [Indirect MR arthrography in the diagnosis of rotator cuff lesions]. AB - PURPOSE: To determine the value of indirect MR arthrography in lesions of the rotator cuff, prospectively versus arthroscopy. METHODS: 63 patients with suspected shoulder pathology were examined: oblique-coronary and axial T1w sequences, axial FLASH-2D sequences, furthermore oblique-coronary T2- and PD weighted sequences were taken. After intravenous administration of 0.1 mmol Gd DTPA/kilogram body weight and active motion of the shoulder T1w sequences were repeated. Signal intensities (SI) inside the tendon were quantitatively measured by the ROI technique (region-of-interest) and the percentual contrast-enhancement CE was calculated. In 32 patients the results were confirmed by surgical follow up. RESULTS: The mean SI measured in the supraspinous tendon were higher in lesions (degeneration, impingement, partial and total rupture), before as well as after contrast medium, compared to intact findings (p < 0.05). Similarly the percentual CE (p < 0.05) was higher. We detected 9 total ruptures and three impingements (for both sensitivity before/after Gd-DTPA 100%), furthermore 6 partial ruptures. Of the latter three were false positive and one false negative, leading to a sensitivity of 75% and a specificity of 50%. CONCLUSION: The visualization of rotator cuff lesions can be optimized with indirect MR arthrography. The detection of partial ruptures remains faulty, because the exact demarcation of degenerative change and partial rupture is still difficult. PMID- 11013612 TI - [Multislice spiral CT of the paranasal sinuses: first experiences using various parameters of radiation dosage]. AB - PURPOSE: To evaluate the potential of multislice CT in the diagnosis of the paranasal sinuses. METHODS: Dose measurements were performed exposing an Alderson Rando phantom in a four-slice spiral CT (MS-CT) while applying a variety of scan parameters. Additionally, 30 consecutive patients underwent a transversal examination by the MS-CT using 1/1/3.5/0.5 mm spiral parameters and an additional transversal or coronal scan on a conventional single slice spiral CT (SS-CT) with 2/3/1 mm. Coronal reformations of the MS-CT were compared with the primary coronal SS-CT, or coronal reformations of the transversal SS-CT, respectively, with regard to image quality and depiction of relevant anatomical details of the region. RESULTS: Superficial exposure values at the level of the eye lenses as well as for the thyroid gland were superior for MS-CT (3.62 mGy, and 0.12 mGy, resp.) as compared to SS-CT (2.96 mGy, and 0.07 mGy). Image quality was equal or superior for MS-CT as compared to SS-CT in all but one case. Drawbacks of SS-CT, such as dental amalgam artifacts, stair step artifacts or partial volume artifacts did not notably affect the coronal reformations of MS-CT. CONCLUSION: MS-CT seems to have the potential to replace primary coronal CT of the paranasal sinuses without any loss of image quality, but may even improve the overall diagnostic value. Radiation doses may still have to be reduced. PMID- 11013611 TI - [MRI-guided percutaneous radiofrequency ablation of hepatic neoplasms--first technical and clinical experiences]. AB - PURPOSE: To evaluate the safety and efficacy of MR-guided radiofrequency ablation (RFA) of hepatic neoplasms. MATERIAL AND METHODS: 16 hepatic neoplasms (1.3-3.0 cm in diameter) in 11 patients were treated by 22 percutaneous RFA sessions during a prospective study. 16 G, MR-compatible cooled-tip electrodes with active lengths of 2 cm and 3 cm, respectively, were placed under MR-guidance in an open 0.2 Tesla MR system (Magnetom Open, Siemens, Erlangen) using fast T1-weighted sequences. Pretreatment studies, evaluation of tumor necrosis (one week after last RFA), and further follow-up studies every 3 months were performed using 1.5 Tesla MR systems. RESULTS: The mean procedure time was 2.8 (1.5-3.3) h. Complications related to percutaneous treatment were not encountered. 14 of 16 neoplasms (87%) showed no CM enhancement during MRI after the last RFA and were judged to be completely necrotic. In 11 tumors one treatment session was necessary, in 4 tumors two and in one tumor three. Follow-up studies revealed persistent complete necrosis in 13 of 14 (93%) tumors during a period of 3-18 (median: 11.8) months. In 5 patients new intrahepatic tumors developed that were not suitable for further RFA treatment because of their number, size and location. CONCLUSION: MR-guided RFA using single cooled tip electrodes is safe and technically effective for treatment of hepatic neoplasms up to 3 cm in size, however further improvements are necessary. PMID- 11013613 TI - [Examination of the image quality from digital fluoroscopy equipment with 1k- and 2k-image matrix]. AB - PURPOSE: The image quality of 1k- and 2k-radiographs from digital fluoroscopy equipments (Sireskop SX and Polystar SX--Siemens) are characterized by comparison with each other to evaluate the relevance of this technique in clinical routine. MATERIAL AND METHODS: We examined fabricated and evaluated images with 1k- and 2k image matrix from several high and low contrast phantoms and from skeleton phantoms. On the one hand, the digital image values can be used directly for the evaluation, on the other hand the comparing evaluation by experienced radiologists resulted from a visual consideration in a blind study. RESULTS: The quality difference of the 1k- and 2k-images depends mainly on the distance of the investigated sector of the object to the image intensifier and on the scattering of the radiation in the object positioned between the investigated sector and the image intensifier. The nearer the investigated object is located to the intensifier and the smaller the radiation is scattered in the object, the more the image quality of a radiograph with a 2k-matrix is increasing in comparison to an image with 1k-matrix. The higher the tube voltage, the smaller are the differences. CONCLUSION: The image quality enhancement because of the more sensitive sampling of the Saticon target in the 2k-matrix is limited by the opening of the iris positioned in the light distributor. Therefore the image quality differences of medical 1k- and 2k-radiographs in many cases are small. PMID- 11013614 TI - [Various forms of rachitis with unusual, malignancy simulating a brown tumor]. PMID- 11013615 TI - [Compression of the suprascapular nerve by a ganglion--MRI diagnosis]. PMID- 11013616 TI - [Posttraumatic dissection of the renal artery--imaging diagnosis and interventional therapy]. PMID- 11013617 TI - [Stent graft implantation in femoral aneurysms with continuous vascular ultrasonography]. PMID- 11013618 TI - [On: "Acute heart arrest in spiral CT" by A. Stoger, B. Munsterer and A. Schinnerl, Innsbruck. in: Fortsch Rontgenstr 2000; 172:490=491]. PMID- 11013619 TI - Trigeminal neuralgia: pathogenesis and treatment. PMID- 11013620 TI - Subtotal petrosectomy in the treatment of cerebrospinal fluid fistulae of the lateral skull base. AB - Cerebrospinal fluid (CSF) fistulae almost invariably lead to meningitis, even in the absence of other clinically obvious sequelae of the fistula such as a CSF fluid leak. The only effective means of reducing the risk of meningitis is surgical closure of the fistula. If surgery is to be recommended to patients with CSF fistulae even if they are currently asymptomatic, the morbidity of the procedure must be a principal determinant of the chosen technique. Recovery after the extracranial approach to a CSF fistula is much more rapid than after an intracranial procedure. The extracranial route is also free of the long-term risk of epilepsy which accompanies a craniotomy. The principal disadvantage of the lateral extracranial approach, failure of treatment, has been largely eliminated following studies into the obliteration of simple bony cavities using free adipose grafts. This paper describes our use of the extracranial approach to closure of CSF fistulae of the lateral skull base. PMID- 11013621 TI - Meningiomas in childhood and adolescence: a report of 13 cases and review of the literature. AB - Meningiomas are relatively uncommon in childhood. They represent 1 to 2% of all intracranial tumours of infancy and childhood. During the last 30-year period, from 1964 to 1993, 13 children with a diagnosis of meningioma were operated on at the Department of Neurosurgery, Hacettepe University School of Medicine. The meningiomas in the paediatric age group were characterized by multiplicity (23%), frequent association with neurofibromatosis (23%), intraventricular location (8%), and the presence of a cystic component in the tumour tissue (23%). These findings in children differ from those in adults. This study constitutes a retrospective analysis of these cases and a review of the literature. PMID- 11013622 TI - Is the technique of posterior transarticular screw fixation suitable for rheumatoid atlanto-axial subluxation? AB - The rheumatoid patient with atlanto-axial subluxation presents a major challenge to the spinal surgeon, owing to the poor wound healing and bone quality. Traditional wiring techniques are associated with a high complication and failure rate. Posterior transarticular screw fixation of the atlanto-axial joint offers a credible alternative and when combined with a Gallie construct offers immediate true 3-point stability. It is, however, a difficult and demanding technique which carries a risk of vertebral artery, cranial nerve and spinal cord damage. The question that arises therefore is "Do the improved stability rates afforded by this technique really justify the risks of arterial and neurological damage?" To date there have been no studies of this technique dealing solely with the rheumatoid patient, with most reports dealing with a heterogeneous patient population, mainly trauma-related cases. The purpose of this report is to analyse critically our results with particular reference to the complications that we have encountered and the technical reasons for their occurrence. We analysed the clinical and radiological data of 38 rheumatoid patients (six males: 32 females, mean age of 54 years) with atlanto-axial subluxation who underwent transarticular screw fixation. Our analysis centred on screw malposition and complications. Parametric and non parametric statistical analysis was performed. Significance was accepted at the 5% level (p < 0.05). Our analysis revealed that three vertebral arteries were damaged. Two of these were recognized at the time of surgery, with the remaining case only suspected following postoperative CT to assess screw positioning. Vertebral artery occlusion was subsequently confirmed by angiography. All three patients were asymptomatic from their arterial injury. There was only one neurological complication in this series, and this was caused by a high screw, which damaged the hypoglossal nerve with a temporary nerve palsy ensuing. Four screws broke, all were made of titanium, but more importantly, all were also associated with contralateral screw malposition. Stability was achieved in 95% of cases overall. The high stability rates afforded by this technique do appear to justify the inherent risks of this procedure. If unilateral screw fixation only is achieved, we would recommend a period of halo immobilization until osseous union occurs. PMID- 11013623 TI - Synovial cysts of the lumbar spine: a review. AB - Four cases of synovial cyst (ganglion) arising from the facet joints of the lumbar spine are reported. A typical presenting feature was exacerbation of pain on standing or walking, mimicking vascular claudication. MRI proved in all four cases to be the definitive investigation and surgery the treatment of choice, producing excellent results. PMID- 11013624 TI - On the usefulness of brain PET scanning to the paediatric neuro-oncologist. AB - Positron emission tomography (PET) scanning adds a functional dimension to brain scanning; levels of metabolic activity are imaged and this information may complement the more "anatomic" imaging of CT and MRI. In a series of 10 young patients the usefulness of PET scanning technique was investigated. The major areas of usefulness were the distinction of posttreatment sequelae from active tumour (both postsurgical MRI changes from tumour and postradiation MRI changes from tumour) and the localization of persisting tumour amenable to radiosurgical treatment. The technique was beneficial in assessing continuing activity in pineal tumours (residual pineal teratoma mass and residual pineocytoma mass) and in assessing activity in brainstem/cerebellar peduncle gliomas (three cases). In one unusual case of widespread leptomeningeal melanoma, the PET scan under-read the situation. PMID- 11013625 TI - Intraventricular craniopharyngioma: a long-term follow-up of six cases. AB - Intraventricular craniopharyngiomas are rare tumours. They are wholly within the third ventricle and can be distinguished from suprasellar lesions which extend into the third ventricle by the presence of an intact floor of the third ventricle. They are attached to the wall of the third ventricle to a variable extent, most commonly in the region of the tuber cinereum. The long-term follow up on six cases treated by one of the authors is presented. Headache and visual disturbance were the most common presenting features but, unlike the more common suprasellar lesions, symptomatic endocrine disturbances were not a common presenting feature. Total surgical removal can cause hypothalamic damage and the resulting morbidity can be serious and sometimes life-threatening. Subtotal removal followed by radiotherapy is probably the treatment of choice for these lesions. PMID- 11013626 TI - Treatment of ependymomas. Clinical and non-clinical factors influencing prognosis: a review. AB - We review the epidemiological and clinical features of ependymomas as described in published series as well as the effect on outcome of various treatment strategies. PMID- 11013627 TI - Fatal tumoural haemorrhage following decompressive craniectomy: a report of three cases. AB - Three cases of large and deep seated anaplastic cerebral glioma were treated by bone and dural decompression. The patients worsened suddenly within 12 h of surgery and later died. Postmortem examination revealed a large intratumoural clot in each case. The effects of decompression and the probable causes of fatal bleeding are analysed in this report. PMID- 11013628 TI - Use of the Gigli saw in performing a mid-frontobasal or pterional craniotomy. AB - The use of the Gigli saw to deal with the inner bony ridges encountered in a mid frontobasal or pterional craniotomy is presented. Despite the fact that power driven craniotomes have certain advantages, the authors favour the Gigli saw in this situation. PMID- 11013629 TI - Free-lying ectopic meningioma within the orbit. AB - Intraorbital ectopic meningiomas, which in most cases occur along the orbital wall, are rare. The authors report a 27-year-old woman who suffered from an intraorbital meningioma lying apart from the optic nerve and having no connection with the periorbita or any small nerves. PMID- 11013630 TI - Recurrent self-inflicted craniocerebral injury: case report and review of the literature. AB - Self-inflicted craniocerebral injuries have been reported exclusively in mentally disturbed patients and criminals. We report a 28-year-old man with a severe mental disorder who initially hammered a nail into his brain and subsequently repeatedly inserted foreign objects into his brain. The literature is reviewed and the surgical and psychiatric management discussed. PMID- 11013631 TI - Case report: inflammatory pseudotumour of the spine, with literature review. AB - A case of inflammatory pseudotumour of the thoracic spine is reported. There appears to be only one previous report of a spinal case. The patient, a 58-year old woman, presented with symptoms and signs of spinal cord compression indistinguishable from those due to extradural tumour. The radiological, operative and pathological findings are presented together with a literature review. PMID- 11013633 TI - Supratentorial haemangioblastoma: appearances on MR imaging. AB - Haemangioblastoma is a rare, benign tumour of vascular origin which usually occurs in the posterior fossa. Supratentorial haemangioblastomas are exceptionally rare. In this report we present three cases of supratentorial haemangioblastoma with MRI findings. PMID- 11013632 TI - Acute bilateral extradural haematoma of the posterior cranial fossa. AB - Traumatic haematomas located in the posterior fossa are less frequent than those above the tentorium. Extradural haematomas are the most common type of haematomas in the posterior fossa and are usually unilateral. We present the case of a patient with a bilateral extradural haematomas of the posterior fossa and review eight cases previously reported in the literature. PMID- 11013634 TI - Bitemporal hemianopia caused by metastatic carcinoma in a patient with Wegener's granulomatosis: an unexpected finding. AB - Metastatic lesions in the suprasellar region are extremely rare. The differential diagnosis of a focal lesion at this site is wide ranging and lesions often have a similar radiological appearance. We present a woman with known Wegener's granulomatosis who lost vision while on medical treatment for a presumed granuloma at this site. Exploration revealed a metastatic carcinoma of unknown origin. Attention is drawn to the importance of making a histological diagnosis in cases of suprasellar lesions. PMID- 11013635 TI - Symptomatic hypophyseal granular cell tumour: endocrinological and clinicopathological analysis. AB - We report a case of a hypophyseal granular cell tumour (GCT) presenting with visual failure and hyperprolactinaemia (serum prolactin level, 274 ng/ml; normal, < 10). Magnetic resonance images demonstrated an intrasellar mass with anterosuperior extension. As the patient had chronic renal failure (CRF) and hyperprolactinaemia is frequent in CRF patients, a firm preoperative diagnosis of prolactinoma could not be made. Transsphenoidal removal of the tumour resulted in improvement of both vision and serum prolactin. Histopathological analysis of the surgical specimen revealed GCT. A cell kinetic study, the first such report for this type of tumour, revealed a relatively high Ki-67 staining index of 3.2%. On electron microscopy, numerous intracytoplasmic granules with various electron densities were demonstrated. Moreover, cell-processes extending from the granule rich cytoplasm contained intracytoplasmic filaments but few granules, suggesting that the filament-rich cells, which are occasionally seen in GCT tissues, are essentially identical to the granule-rich cells in origin. PMID- 11013636 TI - Primary intracranial extradural hydatid cyst extending above and below the tentorium. PMID- 11013637 TI - Stereotactic radiotherapy for arteriovenous malformations. PMID- 11013638 TI - Trepan and trephine. PMID- 11013639 TI - Medicine under socialism: reflections on neurosurgery in the Ukraine. PMID- 11013640 TI - The intensive care of severe head injury: a survey of non-neurosurgical centres in the United Kingdom. AB - Few data exist regarding the management of severe head injury in non neurosurgical centres within the UK. We aimed to discover the number of intensive care units admitting head injury patients, the number of patients admitted annually, and the monitoring and treatment methods followed. Questionnaires were sent to the senior nurse and consultant in 263 intensive care units within non neurosurgical hospitals. The response rate was 78.8%, with at least one response received from 93.2% of hospitals. The severely head injured were routinely admitted in 56.7% of units. Approximately 2100 patients are admitted annually, a mean of 15 per unit. Intracranial pressure monitoring is routine in only 9% of units and 7% are without 24-h facilities for CT, a cause for concern. More encouragingly, 63% of hospitals have access to rehabilitation facilities. Distribution of guidelines to all intensive care units participating in the care of head injury may improve management and outcome. PMID- 11013641 TI - Brain abscess and hereditary haemorrhagic telangiectasia: a report of three cases. AB - Hereditary haemorrhagic telangiectasis (HHT) has long been viewed as a rare disease which is not usually associated with high morbidity and mortality. We report three patients with HHT who developed cerebral abscesses secondary to pulmonary arteriovenous malformations (PAVM). PMID- 11013642 TI - Dynamic MRI of cerebrospinal fluid flow in endoscopic percutaneous ventriculostomy. AB - Endoscopic percutaneous ventriculostomy (EPV) is a new technique to restore the flow of the cerebrospinal fluid (CSF) to the basal cistern in patients with non communicating hydrocephalus (NCH). Cardiac gated MRI techniques have been applied to demonstrate normal and abnormal CSF flow in humans. The aim of this study was to evaluate the ability of a new multislice spin echo dynamic MRI technique and amplitude and phase reconstruction to demonstrate the impairment of CSF flow through the aqueduct preoperatively and to assess the patency of the ventriculostomy after surgery. Thirteen patients with NCH were studied with dynamic MRI before and after EPV using multiple sagittal multislice gated SE acquisitions to demonstrate the CSF flow. Nine patients were clinically improved by EPS, two remained unchanged and two deteriorated. The permeability of the ventriculostomy was confirmed in all patients. We think that MRI flow studies are useful to demonstrate the site of impairment of CSF flow and its aetiology preoperatively, and to assess the permeability of the ventriculostomy after surgery in a non invasive way. PMID- 11013643 TI - Persistent facial pain following microvascular decompression of the trigeminal nerve. AB - Pain in the face following microvascular decompression (MVD) can be due to persisting trigeminal neuralgia (TGN) or a variety of other facial pain syndromes. If magnetic resonance tomoangiography (MRTA) indicates continuing vascular compression and the patient has true persistent TGN, then the patient can be relieved of pain by repeating the MVD. When the MRTA is negative for continuing compression alternative techniques may be employed; section of the nerve at the pons may be the treatment of choice for true persistent TGN in the absence of neurovascular compression. In some cases the pain is dysaesthetic in nature and not persistent TGN. This is always associated with previous destructive lesions to the nerve, usually radio-frequency thermocoagulation. When this component to the pain is recognized pre-operatively the patient must be warned not to expect relief of this same component of the pain from MVD. When it is not possible to classify the facial pain clinically, improvement does not occur following MVD even when there is clear evidence of vascular compression on MRTA. PMID- 11013644 TI - Treatment by a specialist surgical neuro-oncologist does not provide any survival advantage for patients with a malignant glioma. AB - To determine whether patients with malignant glioma (glioblastoma and anaplastic astrocytoma) had longer survival times, and lower morbidity and mortality if operated on by a surgical neuro-oncologist rather than a general neurosurgeon the outcomes of 236 patients managed within a university surgical neurology department were analysed. Although both surgical morbidity (8.9 versus 11.8%) and mortality (3.6 versus 8.8%) were lower following surgery by the specialist neuro oncologist neither difference was statistically significant. Crude outcome data suggested patients operated upon by a specialist surgical neuro-oncologist survived longer (p = 0.067). However, after adjustment for case mix (type of tumour, year of treatment, MRC prognostic index) using multiple logistic regression and a hazards model, there was no difference in outcome (p = 0.46, HR 0.884, 95% CI 0.639-1.22). This retrospective study (i) suggests that other outcome measures are required to validate specialist surgical neuro-oncologist treatment of patients with malignant glioma; and (ii) confirms the importance of adjustment for case mix when comparing non-randomized treatment outcomes. PMID- 11013645 TI - Dynamic CSF flow study in the pathophysiology of syringomyelia associated with arachnoid cysts of the posterior fossa. AB - Two patients with posterior fossa arachnoid cysts associated with syringomyelia are discussed adding to the five cases already reported in the literature. The formation and progression of syringomyelia secondary to a posterior fossa cyst and its possible pathophysiology by dynamic CSF flow studies using magnetic resonance (MR) imaging are discussed and reviewed. PMID- 11013646 TI - Factors implicated in deaths from subarachnoid haemorrhage: are they avoidable? AB - The major causes of death following subarachnoid haemorrhage are the effects of the initial bleed, aneurysmal rebleeding and delayed cerebral ischaemia. Although in many cases the causes are unavoidable, in others they are potentially preventable. By conducting a Regional Audit of patients who have died from subarachnoid haemorrhage, we have attempted to quantify these concerns. The medical records and CT scans of 200 patients who died from subarachnoid haemorrhage in hospitals in the East Anglian Region over a 5-year period were analysed, with particular regard to the identification of potentially avoidable factors, including radiologically confirmed rebleeding, marked hydrocephalus, and poor fluid and electrolyte resuscitation. Thirty-three patients presented with World Federation of Neurosurgical Societies (WFNS) Grade V with fixed pupils. These patients were deemed unsalvageable. Of the remainder, 106 were good grade (WFNS I-III) at initial presentation, 77 (73%) of whom died as a result of rebleeding (mean 10.4 days post-bleed) indicating delays in transfer and definitive treatment. Of the 61 poor grade patients (WFNS IV and V with reactive pupils) marked hydrocephalus occurred in 15 (25%) and poor resuscitation in 37 (61%). These were considered contributing factors to their poor clinical condition. Rebleeding was the main cause of death in the good grade patients, and correctable factors were identified which are known to contribute to a poor clinical condition in poor grade patients. The need for more rapid and active clinical intervention is indicated. PMID- 11013647 TI - Seizures following posterior fossa surgery. AB - A retrospective analysis of seizure incidence in 511 patients who underwent posterior possa operations via a suboccipital craniectomy with prophylactic anticonvulsant agents, was performed. Thirty patients (5.9%) experienced seizures within 2 weeks postoperatively. Their mean age was 15 years. Twenty patients had seizures within 3 h of operation; 24 patients had generalized tonic clonic seizures. Focal motor seizures with secondary generalization and simple focal motor seizures were recorded in three patients each. The incidence of postoperative seizures was highest in patients with acoustic schwanommas (9.09%) followed by those with medulloblastomas (8.54%) and astrocytomas (8.33%). The sitting position, associated with venous air embolism (VAE) and or pneumocephalus, was related to the occurrence (p = 0.001) of postoperative seizures. Seizures occurred in 24 patients out of 250 cases operated on in the sitting position compared with 3/170 and 3/91 in the prone and lateral positions, respectively. Intraoperatively significant VAE occurred in 10 out of 30 patients and postoperative computed tomography revealed pneunocephalus in 20 out of 30 patients. A higher percentage was found in patients with a preoperative ventriculoperitoneal shunt or intraoperative ventriculostomy (6.5%) than in those without (5.1%), but the difference was not statistically significant. In conclusion, seizures after posterior fossa surgery are a significant problem which is not reflected in the published literature. Our study highlights the significance of the sitting position in the causation of seizures after posterior fossa surgery. PMID- 11013648 TI - A technique for dural repair following retromastoid suboccipital craniectomy. AB - The authors describe a simple technique used to achieve a water-tight closure of the dura mater during suboccipital craniectomy. PMID- 11013649 TI - Paired colloid cysts of the third and lateral ventricles. AB - We report the case of a man of aged 27 years who presented with obstructive hydrocephalus caused by a colloid cyst of the third ventricle. He was found to have an additional and larger colloid cyst lying adjacent to it, but within the lateral ventricle. The contents of the two cysts were of different consistency. We have been able to find only one previous report of more than one colloid cyst occurring in the same patient and none where one of the cysts lay within the lateral ventricle. PMID- 11013650 TI - Chronic encapsulated intracerebral haematoma in a patient with medically intractable epilepsy. AB - A patient with a chronic encapsulated intracerebral haematoma presenting with medically intractable epilepsy is described. A tough capsule containing an old haematoma was confirmed surgically, and consisted of dense collagenous tissue with rich neovascularization. The radiological features, aetiology and treatment of this rare occurrence are discussed. PMID- 11013651 TI - Giant cephalhaematoma in a 17-year-old boy with Ehlers-Danlos syndrome. AB - We report a case of giant cephalhaematoma in a 17-year-old boy with Ehlers-Danlos syndrome. This haematoma occurred after a minor head injury. It increased in size, immediately after needle aspiration and was responsible for considerable blood loss. Possible physiopathological mechanisms and treatment modalities are discussed. PMID- 11013652 TI - Intraventricular and leptomeningeal dissemination of a pilocytic cerebellar astrocytoma in a child with a ventriculoperitoneal shunt: case report. AB - Dissemination of a pilocytic cerebellar astrocytoma is a very rare occurrence. So far only eight cases have been reported in the literature and in only one of these cases had the tumour spread into the ventricles. We report a case of a child who presented with communicating hydrocephalus and a small cerebellar lesion. The patient was initially treated by a ventriculoperitoneal (VP) shunt and the lesion was followed-up. Two years later, intraventricular and leptomeningeal dissemination of the tumour which proved to be a pilocytic astrocytoma was documented. The role of the VP shunt in diverting metastasizing tumour cells into the ventricles is discussed. PMID- 11013653 TI - Low grade pilocytic astrocytoma presenting as a spontaneous intracerebral haemorrhage in a child. AB - A girl aged 13 years presented with spontaneous intracerebral haemorrhage. Initially, no cause could be found but she was later found to have a low grade pilocytic astrocytoma. This very uncommon presentation was not considered before surgery. PMID- 11013654 TI - Very late cerebral metastasis from malignant melanoma. AB - The appearance of cerebral metastases of malignant melanoma (MM) more than 10 years after the primary diagnosis is extremely rare. We report the case of a patient with a solitary brain metastasis of MM who came to our observation 11 years after the treatment of the cutaneous lesion. This patient, who up until then had appeared disease free, presented with two episodes of intracranial haemorrhage in a 5-month period. Neuroradiological findings (CT, MRI, angiogram) did not suggest a brain metastasis. The correct diagnosis was reached only after histopathological examination of the surgically removed lesion. On the basis of this experience, we stress the importance of a long-term clinical and radiological follow-up of all patients with MM. PMID- 11013655 TI - Spontaneous CSF rhinorrhoea due to temporosphenoidal encephalocele. AB - A 54-year-old woman was referred with spontaneous cerebrospinal fluid (CSF) rhinorrhoea. CT cisternography revealed a defect in the lateral wall of the left sphenoid sinus, with an anteromedial temperosphenoidal encephalocele associated with contrast leakage into the sinus. Subsequent monitoring showed raised intracranial pressure. Transcranial intradural repair of the encephalocele followed by lumboperitoneal shunt for the high pressure abolished the leakage of cerebrospinal fluid. PMID- 11013656 TI - Neuronavigation: where is the evidence and will there ever be any? PMID- 11013657 TI - The MEDUNSA sign. PMID- 11013658 TI - Comment on 'Value of early postoperative plain radiography following BOP fusion in cervical spinal surgery'. PMID- 11013659 TI - Multiple primary cerebral hydatid cysts. PMID- 11013660 TI - Cervical degenerative disease: discectomy or fusion? PMID- 11013661 TI - A survey of the peri-operative management of patients undergoing anterior cervical decompression in the UK and Eire. AB - The peri-operative management of patients undergoing single level anterior cervical decompression (ACD) in neurosurgical departments in the UK and Eire has been surveyed. Eighty-eight per cent (132 of 150) of consultant neurosurgeons responded and of these 118 perform ACD. Nearly all (96%) respondents use interbody grafts and most of these use autologous bone (63%). A small minority (4%) never use interbody grafts. Forty-four per cent of surgeons use the same surgical technique irrespective of the underlying pathology (Cloward 57%: Smith Robinson 33%; ACD without graft 8%). Surgibone grafts are used by 25% and BOP grafts by 5%, but 15% of surgeons have abandoned artificial graft materials because of concerns about non-union. This survey has revealed a diverse range of practice amongst UK neurosurgeons. A prospective randomized trial is required to identify whether the theoretical benefits of interbody grafting following cervical discectomy justify donor site morbidity and the expense and risks of substitute graft materials. PMID- 11013662 TI - The costs of managing patients with malignant glioma at a neuro-oncology clinic. AB - Malignant glioma (glioblastoma and anaplastic astrocytoma) remain incurable despite extensive resection, radiotherapy, chemotherapy and experimental therapies. Few studies have addressed either the costs of various treatments for malignant glioma or their cost effectiveness. The aims of this study were to identify direct hospital costs of treating patients with biopsy proven malignant glioma. The study was carried out within the setting of a dedicated neuro oncology clinic at a university teaching hospital and included 236 patients treated between 1989 and 1995. The study used the unit costing of each item of treatment according to NHS National Costing Project. The cost of treatment was broken down into its various components: bed days, investigations, surgery, radiotherapy, chemotherapy and neuro-oncology out-patient follow-ups. The mean costs for each of the items based on 1995 figures for the 157 patients having surgery followed by radiotherapy were neuroradiological investigations (442 Pounds), neurosurgical bed days (2407 Pounds), neurosurgery (2068 Pounds), neuropathology (434 Pounds), radiotherapy (8832 Pounds), out-patients (1078 Pounds) and chemotherapy (440 Pounds). Total treatment costs per patient ranged from 1978 Pounds to 26,980 Pounds. Median costs of care decreased sequentially with worsening MRC Brain Tumour prognostic group. Management of patients with the best prognosis (MRC index score of 1-10) cost a median of 16,550 Pounds (range 4572-26,090 Pounds) whilst the median management cost of those in the worst prognostic group (MRC score 34-38) was 6514 Pounds (range 1978-18,360 Pounds). The median cost of each week of survival in the patients with the best outcome (MRC score 1-10) was < 150 Pounds compared to 232 Pounds for each week of survival for patients in the worst prognostic group (MRC score 34-38). This study made no attempt to collect costs of supportive or community-based care. Prospective studies are required to collect such data, as well as assessing the costs effectiveness of alternative treatment strategies. PMID- 11013663 TI - Extradural haematoma and diffuse axonal injury in victims of fatal road traffic accidents. AB - Seven cases of victims of road traffic accidents with extradural haematoma (EH) and diffuse axonal injury (DAI) are reported. Such cases are part of a total of 120 victims of fatal road traffic accidents that were subjected to pathological study. The association of the two lesions occurred in 5.8% of the patients and all cases of EH were associated with DAI. The latter explains the immediate coma (absence of lucid interval) and the grave prognosis for all seven patients. It was noted that EH is relatively infrequent in road traffic accidents, but in such cases it is more severe since it is frequently associated with DAI and immediate coma. PMID- 11013664 TI - The microbial spectrum of brain abscess with special reference to anaerobic bacteria. AB - The bacteriological and clinical findings of 18 cases of brain abscess are presented. Identification and antimicrobial susceptibility of anaerobic strains was performed both by conventional methods and the newer RapID ANA II panel and E test methods, respectively. Characterization of the anaerobic isolates was done by restriction fragment length polymorphism (RFLP). Anaerobic or aerobic organisms alone were recovered in three (16.6%), and mixed aerobic and anaerobic in another three (16.6%) patients. There were nine anaerobic isolates. The predominant anaerobes were Prevotella melaninogenicus (four isolates), Bacteroides preacutus (three isolates), Fusobacterium nucleatum and Peptostreptococcus sp. (one isolate each). A total of six aerobic isolates, all of which were Gram-positive cocci were recovered. All the anaerobic isolates were susceptible to metronidazole, but two isolates of P. melaninogenicus were resistant to penicillin. There was complete agreement between the identification based on biochemical profiles and RFLP patterns. These findings indicate the microbial complexity of brain abscess and the need to target antimicrobial therapy against both the aerobic and anaerobic components of infection. PMID- 11013665 TI - The preoperative skin shave in neurosurgery: is it justified? AB - Shaving the scalp prior to surgery is a very common practice. Out of 105 cases operated upon without skin shave at the Royal London Hospital, only one became infected (0.95%). A search into the history of aseptic surgery shows that there are no scientific grounds for the practice of shaving. An examination of contemporary practices worldwide shows that there is a greater realization that preoperative skin shaving does not confer any benefit against postoperative wound infection and that, paradoxically, it may lead to higher rates of wound infection due to the epidermal injury that it inflicts. PMID- 11013666 TI - Patients with increased intracranial pressure cannot be monitored using near infrared spectroscopy. AB - The hypothesis of this study was that with near infrared spectroscopy a decrease of the regional cerebral saturation would be detectable in patients with increased intracranial pressure. For this purpose, two study groups were compared: (A) eight healthy volunteers and (B) eight heart beating organ donors with brain stem herniation. The regional cerebral saturation was measured according to the developer's suggestions. Mean arterial pressure and peripheral pulse oximetry were recorded. The statistical comparison, using the unpaired t test, revealed no significant differences for the regional cerebral saturation (p = 0.97) or for the mean arterial pressure or peripheral pulse oximetry between the groups. In both of the studied situations, no differences in the regional cerebral situation were detectable, even though the patient situations were completely antagonistic. Therefore, it is reasonable to discourage the use of the INVOS 3100 Cerebral Oxymeter to monitor patients with increased intracranial pressure indirectly. PMID- 11013667 TI - A preliminary experimental in vivo study of the effect of photodynamic therapy on human pituitary adenoma implanted in mice. AB - As surgery alone may prove inadequate to effect a cure for invasive pituitary adenomas, photodynamic therapy (PDT) was investigated as a possible adjuvant treatment for this group of tumours. Different subtypes of human pituitary adenoma cells were implanted subcutaneously into nude mice to study the in vivo effect of PDT on such lesions. The photosensitizer used in this study was polyhaematoporphyrin at a dose of 10 mg/kg b.w., followed by light irradiation at a wavelength of 630 nm with varying light doses between 10 and 75 J/cm2. Histopathological examination of the treated implants consistently showed tumour vascular changes with acute inflammatory reaction, interstitial haemorrhage, and evidence of cell death at higher doses of light. These changes were absent in the control groups. These findings indicate that the cytotoxic effect of PDT demonstrated in vitro in previous studies, is also present in vivo. PMID- 11013668 TI - The pallidotomy debate. AB - It has been suggested that image-guidance and macro-stimulation alone are not sufficiently accurate to result in safe and effective lesion localization in pallidotomy for Parkinson's disease when compared with micro-electrode recording. This review analyses the data in the series published to date, and compares the safety and efficacy of the two techniques, finding no evidence to support this claim. In addition, evidence regarding the necessary accuracy of lesion placement is reviewed. PMID- 11013670 TI - Clipping of an aneurysm of a fenestrated basilar artery. AB - We describe the case of a fenestrated basilar artery aneurysm successfully clipped by means of a right presigmoid petrosal approach. Three-dimensional CT angiography enabled the surgeon to plan an approach to an aneurysm in this unique location, and the presigmoid petrosal approach was suitable for clipping of the aneurysm. PMID- 11013669 TI - Brain stem contusion due to tentorial coup injury: case report and pathomechanical analysis from normal cadavers. AB - This report is in two parts. First, a case report on a 20-year-old man with a localized brain stem contusion. Second, in order to elucidate the mechanism of this injury, an anatomical study was performed. Ten cadaver heads were analysed to reveal the variations of spatial anatomy around the tentorial incisura. The lateral tentorial incisura (lateral to brain stem) was situated at the level of pontomesencephalic junction and nearest to the brain stem along its course. The shortest distance between them averaged 1.0 mm (0-4 mm). Based on these findings, primary brain stem injury caused by tentorial incisura occurs at its lateral portion due to the shortest distance to the brain stem and near the level of pontomesencephalic junction. In patients with a tentorial incisura closely related to or touching the brain stem, tentorial coup injury to the brain stem may occur even with a relatively minor injury. In our case, repeated CT and MRI proved that the location of contusion was at the pontomesencephalic junction, coinciding with the level of the tentorial edge. The injury started at the surface of brain stem. The tentorial edge was close to brain stem in this case. These radiological findings support the hypothesis that the brain stem contusion was caused by a tentorial coup injury. PMID- 11013671 TI - Craniofacial fibrous dysplasia complicated by acute, reversible visual loss: report of two cases. AB - We report two cases of craniofacial fibrous dysplasia which presented with acute visual loss. The first patient had a sphenoid sinus mucocele compressing the optic chiasm. In the second patient the optic canal was narrowed by dysplastic bone. In both cases optic nerve decompression restored vision to normal. PMID- 11013672 TI - Adynamic ileus complicating lumbar laminectomy: a report of two cases. AB - Adynamic ileus is a well recognized complication of spinal injuries and disease. We report two cases of adynamic ileus affecting both small and large bowel following uncomplicated L4/5 laminectomy. Both patients recovered with conservative treatment, but their hospital stay was prolonged. We have found three other cases in the literature of ileus affecting only the colon following laminectomy at this same level. PMID- 11013673 TI - Primary cranial Ewing's sarcoma. AB - Primary Ewing's sarcoma of the cranium is extremely rare, with only 17 cases reported so far. We describe a further case with involvement of the frontotemporal region and the orbit. The patient, an adult male, was admitted with headache and papilloedema, and later developed localised swelling and proptosis. He was treated with surgery, radiotherapy and chemotherapy, and was free from metastases during a follow-up of 14 months. The prognosis of Ewing's sarcoma is improving with radiotherapy and chemotherapy. Further cases are needed to study the biological behaviour of primary cranial Ewing's sarcoma. PMID- 11013674 TI - Brain abscess as the presenting feature of melioidosis. AB - Central nervous system involvement in melioidosis is rare and there are only a few reports of the causative organism, Burkholderia pseudomallei, causing a brain abscess. We report a patient who presented to us with a brain abscess due to this organism and emphasize the need for a high degree of suspicion for this disease in tropical countries and treatment with the appropriate antibiotics, as the mortality associated with this disease is very high. PMID- 11013675 TI - Anterior skull base osteoid osteoma: case report. AB - Osteoid osteoma is a benign bone tumour which rarely occurs in the skull. A case of such a tumour in the posterior ethmoid region bulging into the anterior cranial fossa is reported in a 42-year-old woman who presented with intense frontal headaches. Excision of the tumour was performed through a subfrontal approach. PMID- 11013676 TI - Unilateral proptosis and chemosis caused by dural arteriovenous malformation of the superior sagittal sinus. AB - We describe a patient with unilateral proptosis and chemosis resulting from a dural arteriovenous malformation (AVM) of the superior sagittal sinus (SSS) fed mainly by branches of both external carotid arteries. The symptoms may have been caused by increased SSS pressure and disturbance of venous flow by the dural AVM. PMID- 11013677 TI - Primary spinal melanoma: case report. AB - We report a case of primary melanoma of the thoracic spinal cord revealed by progressive bilateral lower extremity weakness associated with sensory loss and urinary dysfunction. The preoperative MRI revealed an intramedullary tumour from T7 to T9. Treatment was by complete surgical excision without radiotherapy. Histopathology and immuno-histochemical studies confirmed the diagnosis. The postoperative course was satisfactory with no sign of recurrence after 28 months of postsurgical follow-up. Primary spinal melanomas are rare intramedullary tumours that can be cured by appropriate surgical treatment. PMID- 11013678 TI - Trochlear nerve neurinoma. PMID- 11013679 TI - Medical editors trial amnesty (META) PMID- 11013680 TI - Sir Geoffrey Jefferson 1886-1961. PMID- 11013681 TI - Distal anterior cerebral artery aneurysms: a clinical series. AB - Thirty patients with distal anterior cerebral artery (DACA) aneurysms were seen at the Royal Adelaide Hospital in the period 1970-1996. There were seven males (23%) and twenty three females (77%) with a mean age of 50 years. The average follow up was 5 years. Multiple aneurysms were present in seven cases (23%). The mean size of aneurysms was 5 mm. There were two post-traumatic aneurysms and one mycotic aneurysm. Out of the 30 cases, 19 presented with subarachnoid haemorrhage from ruptured DACA aneurysms. Eight (42%) of them were in good clinical grade (I or II). Operations were carried out in 25 (83%) patients. All five cases with unruptured aneurysms and the eight patients with good clinical grade had good recovery. In contrast, only six (55%) out of 11 patients with poor clinical grade had good outcome. The overall management mortality for the 19 cases with ruptured aneurysms was 16%. Postoperative complication occurred in two cases (8%), one patient developed deep vein thrombosis and seizures, the other patient had transient upper limb weakness. Although there is a definite trend towards better management outcome in the published series of DACA aneurysms over the years, there is still significant mortality and morbidity in the poor grade patients. Early surgery will prevent the deaths from rebleeding and may allow optimal management of vasospasm. PMID- 11013682 TI - Histopathological changes following the use of biological and synthetic glue for dural grafts: an experimental study. AB - Dural defects and CSF leaks are common neurosurgical problems and the search for a suitable sealant continues. The effects of such sealants on meninges and brain have not been previously studied. To study the effect of two commonly used sealants, burrholes were made in 20 rats, with the creation of dural tears which were covered with human cadaveric dura smeared with synthetic (cyanoacrylate) glue and gelatine (GRG) glue. Plain dura, placed in one of the burrholes served as a control. An intense inflammatory reaction was observed with both glues, which was found to persist till the eleventh week. PMID- 11013684 TI - A clinical evaluation of the Codman MicroSensor for intracranial pressure monitoring. AB - The strain-gauge Codman MicroSensor intracranial pressure (ICP) transducer has shown consistently good laboratory performance. To assess the practical performance of the system in patients following acute brain injury, 10 patients were fitted with a MicroSensor and a second ICP monitor. In five cases this was a fibre-optic transducer and in five cases an intraventricular fluid-filled device. Paired ICP values were recorded every 5 min. ICP values ranged from 0 to 31 mmHg. Altman-Bland plots showed that individual readings could differ by as much as 9 mmHg. Further analysis showed that much of this disagreement could be explained by a constant offset on each occasion. Comparison traces of ICP in individual patients show high agreement in timing and size of changes. The unexplained constant offset leads to uncertainty about the true ICP. Treatment decisions are often based upon absolute levels of ICP and patient care may therefore differ depending upon the monitor used. PMID- 11013683 TI - Chemotherapy for aggressive or anaplastic high grade oligodendrogliomas and oligoastrocytomas: better than a salvage treatment. AB - This series reports a pilot study of chemotherapy conducted for newly diagnosed and relapsing patients with oligodendrogliomas and oligoastrocytomas. Twenty three patients with clinically or histologically aggressive tumours were eligible to receive procarbazine CCNU and vincristine (PCV) chemotherapy followed by radiotherapy in previously non irradiated patients. Sixteen (69%) responded to chemotherapy with complete responses in two patients and partial responses in 14. Previously irradiated patients were as likely to respond to PCV as those previously non irradiated. An over 1-year history of seizures was the main clinical prognostic factor of response. All toxicities were manageable and no treatment related deaths occurred. Chemotherapy is an effective treatment in aggressive oligodendrogliomas. Further studies must assess the role of chemotherapy in the multidisciplinary management of oligodendroglioma. PMID- 11013685 TI - Benign tumours of the pineal region: a prospective study from 1983 to 1997. AB - Tumours of the pineal region are rare. Between 1983 and 1997, 128 patients with pineal masses were treated in our institution. Forty-eight (38%) of these were benign. There were 13 patients with meningiomas, 11 with epidermoid tumours, 10 with cystic lesions, five with vascular lesions, five with infective pathology and four with mature teratomas. All patients were managed surgically. Ventriculo peritoneal shunts were inserted preoperatively to relieve hydrocephalus. Open surgery thereafter was mostly done through the supracerebellar infratentorial approach. The occipital transtentorial route was preferred for meningiomas. Stereotactic biopsy was used in two patients only. Radical excision of tumours was achieved in 55% of operated cases, partial excision in 25%. Radiation therapy, which was the mainstay of treatment earlier, was used only in two patients. Stereotactic radiosurgery was used in one patient. All patients followed up were found to be in good or excellent condition. PMID- 11013686 TI - A study on the efficacy of intraventricular urokinase in the treatment of intraventricular haemorrhage. AB - Twenty-one patients with intraventricular haemorrhage were randomized to two treatment groups. Both groups had bilateral external ventricular drains inserted, but only the treatment group received 50,000 IU urokinase instilled into the ventricles. The clinical and radiological progress, and 1- and 6-month outcomes were compared. The group that received urokinase treatment was shown to have an improved outcome, with a lower mortality and a lower incidence of hydrocephalus requiring shunt insertion. No haemorrhagic complications were seen in either group, although the treatment group had a slightly increased rate of drain related ventriculitis. PMID- 11013687 TI - Gliomatosis cerebri: disconnection of the cortical grey matter, demonstrated on PET scan. AB - 18-FDG and 11C methionine PET scans were performed on two patients with gliomatosis cerebri. The cortical grey matter was hypometabolic when compared with normal. The findings support the concept that the cerebral cortex becomes functionally disconnected in this disease owing to the infiltrative nature of the underlying tumour. This may account for the high incidence of dementia in the course of this disease. In one of the cases described here, there was clear evidence of progression from a discrete tumour mass of glioma to gliomatosis cerebri and this progression argues against the WHO classification of this disorder separately from glioma. PMID- 11013688 TI - The role of descending basal ganglia connections to the brain stem in parkinsonian akinesia. AB - Akinesia is the most disabling symptom of Parkinson's disease. The neural mechanisms underlying it probably involve the descending projections of the basal ganglia to the brain stem as it improves after a pallidotomy or subthalamic nucleotomy but not after a thalamotomy. We describe the effects of lesioning the pedunculopontine nucleus in the normal primate in generating an akinetic syndrome. The possible clinical implications of this study are discussed. PMID- 11013689 TI - Are there differences in the symptoms, signs and outcome after lumbar disc surgery in the elderly compared with younger patients? AB - Age is claimed by several authors to be a predictor of bad outcome after lumbar disc surgery. The aim of this study is to evaluate the influence of the patients age on preoperative symptoms and the outcome. General data, symptoms, signs and neurological findings of 219 patients were recorded preoperatively Ninety-two per cent of the patients received a follow-up examination by an independent investigator after a mean of 298 days. All patients older than 59 years were defined as belonging to the group of elderly patients (n = 30). The other patients were defined as the younger group (n = 189). The outcome was measured by a visual analogue rating scale. Outcome was not statistically different in both groups. In our opinion, the indications for surgery for a lumbar disc herniation should be no different for older patients even though nerve root tension signs appear to be less marked in older patients. PMID- 11013690 TI - Multilayer reconstruction of the anterior cranial fossa floor. AB - A case is described where a reconstruction of a complex defect in the floor of anterior cranial fossa was successfully carried out using multiple layers of vascularised pedicle flaps. The 42-year-old male patient had previously undergone multiple surgical procedures for persistent cerebrospinal fluid (CSF) rhinorrhoea following a vehicular accident 16 years previously. The extensive vascular supply to the layers of the scalp, pericranium, temporalis muscle and its fascial coverings was exploited in fashioning the flaps. PMID- 11013691 TI - Neurenteric sinus dorsal to the thoracic spine with an associated cutaneous haemangioma: case report. AB - The case of an infant born with a large cutaneous haemangioma overlying the thoracic spine is presented. A small midline pit lay within the haemangioma, histological examination of which confirmed it to be a neurenteric sinus. The possible embryological origins of this rare lesion are discussed. PMID- 11013692 TI - Primary sphenoid and petrous apex esthesioneuroblastoma: case report. AB - A 62-year-old woman presented with raised intracranial pressure and features of a right cerebellopontine angle tumour with extension into the right middle cranial fossa. The patient died before a surgical excision could be performed. The autopsy revealed a primary esthesioneuroblastoma of the sphenoid sinus eroding the petrous bone and extending into the middle cranial fossa with metastatic tumour in the liver, and paratracheal and hilar lymph nodes. Although rare, esthesioneuroblastoma must be considered in the differential diagnosis of petrous sphenoid lesions. PMID- 11013693 TI - Intramedullary ossified cavernous angioma of the spinal cord: case report. AB - A case of a heavily ossified cavernous angioma of the spinal cord along with its histological features and surgical implications is reported. The unusually dense calcification and even bone formation along with the unique eggshell-like cyst formation posed problems in diagnosis and surgical excision. This calcified vascular lesion was treated by subtotal excision. PMID- 11013694 TI - Posterior fossa arachnoid cyst presenting as high cervical cord compression. AB - We report a 16-year-old boy who presented with high cervical cord compression caused by a midline posterior fossa arachnoid cyst lying below the vermis. Cystoperitoneal shunting resulted in a dramatic improvement of neurological signs. PMID- 11013695 TI - Pineal apoplexy: an occurrence with no diagnostic clinicopathological features. AB - Symptomatic pineal apoplexy unlike pituitary apoplexy is uncommon. A patient with an apoplectic pineal cyst, identified preoperatively using MRI and confirmed histologically presented with episodic syncope, and features of raised intracranial pressure, but no localizing neuro-ophthalmological signs. This case prompted a review of the clinicopathological features of pineal apoplexy. There are no diagnostic clinical features and the neuropathological associations of pineal region haemorrhage are diverse. There is no consistent clinicopathological syndrome of pineal apoplexy. PMID- 11013696 TI - Anaplastic ganglioglioma: case report and review of the literature. AB - A 10-year-old girl underwent radical decompression of a right temporal tumour extending to the insular cortex which proved to be a differentiated ganglioglioma. One year later, she presented with a bleeding right frontal tumour extending to the insula, the histology of which showed it to be the anaplastic variant of ganglioglioma. She underwent postoperative radiotherapy but died 6 months after the second operation. Anaplastic ganglioglioma is a rare variant of ganglioglioma and progression of the latter to anaplastic ganglioglioma is an extremely rare occurrence. The literature regarding ganglioglioma is reviewed. PMID- 11013697 TI - Multiple cerebellar haemangioblastomas symptomatic during pregnancy. AB - A patient with von Hippel-Lindau disease presented during her seventh week of pregnancy with obstructive hydrocephalus and brainstem compression caused by multiple cerebellar haemangioblastomas. Surgery was performed to relieve the hydrocephalus and remove the right cerebellar tumour. The patient's subsequent course supports the hypothesis of vascular engorgement of cerebellar haemangioblastomas during pregnancy. PMID- 11013699 TI - Trigeminal neuralgia. PMID- 11013698 TI - Pituitary tumours: recommendations for service provision and guidelines for management of patients. Royal College of Physicians. PMID- 11013700 TI - The role of mebendazole in the surgical treatment of central nervous system hydatid disease. PMID- 11013701 TI - [Imaging of conductive hearing loss]. AB - CT study of the middle ear and ossicular chain plays a major role in assessing the etiology of conductive hearing loss. A normal otoscopic examination is compatible with a fenestral otosclerosis, a minor ossicular chain aplasia or a traumatic dislocation of the ossicular chain. Fenestral otosclerosis diagnosis does not usually require a CT examination except in children without familial history and mixed hearing loss with a transmissional component. In retraction pockets and middle ear cholesteatomas CT allows precise location, extension and defines surgical landmarks. In middle ear tumors CT and MR studies need to be used in conjunction to differentiate glomus tumor, primary cholesteatoma and congenital vascular anomalies. PMID- 11013702 TI - [Imaging of post-traumatic tinnitus, vertigo and deafness]. AB - Tinnitus, hearing loss, and more rarely disequilibrium are common sequela of temporal bone trauma. Hemotympanum may cause a transient and immediate conductive hearing loss. HRCT depicts ossicular dislocation (most frequently incus), producing a long-term conductive hearing loss. Labyrinthine trauma causing neurosensory hearing loss or/and acute vertigo may be depicted by MRI, showing an abnormal non-enhancing high signal T1 of the membranous labyrinthine fluid. MRI also may show low signal T1 and T2 fibrotic areas of the membranous labyrinth, especially of interest if cochlear implant surgery is planned. Perilymphatic fistulas are to be searched in case of fluctuant hearing loss. Both HRCT and MRI may show window damage: filling of the tympanic recess, rupture of the window membrane, intra-vestibular luxation of the stapes, or occasionally pneumolabyrinth. PMID- 11013703 TI - [Polyps and polyposis of the paranasal sinuses]. AB - Discussion is about the different types of naso-sinusal polyps, isolated inflammatory polyps and some other ambiguous types like inverted papilloma, so called haemorrhagic polyps, pseudo-polyps and reactional polyps collecting very heterogeneous and various clinical, CT and evolutive entities. PMID- 11013704 TI - [Imaging of tumors and pseudotumors of the ear]. AB - CT and MRI are in most of the cases associated in imaging of tumours and pseudo tumours of the temporal bone. The tumours of the external auditory meatus particularly the malignant ones are rare. It is of a great interest to delineate a possible extension to the chorda tympani. Secondary cholesteatoma are the most frequent pseudo-tumours of the middle ear: CT and MRI evaluate very well their extension particularly in the giant forms. Others tumours like tuberculosis, histiocytosis, primitive cholesteatoma, tympanic body++ tumours may be diagnosed by imaging as well as the neuroma of the facial nerve, the meningioma or some rarer lesions as the adenoma, or the carcinoid tumour of the middle ear. The adenoma of the endolymphatic sac is the only true tumour of the membranous labyrinth whose diagnosis is nicely made by MRI. The primitive cholesteatoma, the neuroma of the facial nerve may reach the labyrinthine bone and are easily diagnosed by imaging which generally speaking is very good in evaluating the kind of pathology of each cavities, delineating the extension of the lesions and orientating the surgical procedures. PMID- 11013705 TI - [Imaging of cervical masses]. AB - Imaging plays an essential role in the management of neck disease. It can identify true disease versus pseudomasses, and it allows to differentiate between lymph node metastases and other cervical masses. MR imaging and/or CT can show the exact location and extent of a lesion, and can help to anticipate its nature. PMID- 11013706 TI - [Radio-anatomy of lymph node groups in the area of otolaryngology]. AB - Imaging plays an essential role in the management of head and neck lymph node. Knowledge of the anatomy is indispensible to interpret ultrasound, CT, and MR examination. PMID- 11013707 TI - First report on the systematic sequencing of the small genome of Encephalitozoon cuniculi (Protozoa, Microspora): gene organization of a 4.3 kbp region on chromosome I. AB - Belonging to a large group of parasitic amitochondrial protozoans (Microspora), Encephalitozoon cuniculi infects humans and other mammals. Because of its medical importance and small genome size (2.9 Mbp), we are systematically sequencing its smallest (217 kbp) chromosome. The shotgun cloning strategy now has produced the sequence of randomly dispersed contigs representing more than 180 kbp of this chromosome. The present report describes analysis of the 4.3 kbp contig, which includes the complete coding regions of dihydrofolate reductase (DHFR), thymidylate synthase (TS), and serine hydroxymethyl transferase (SHMT) genes and the partial coding region of an aminopeptidase (AP) gene. In contrast to the other reported protozoan genes, DHFR and TS are encoded by two different open reading frames (ORFs). The SHMT gene is the first one identified in a protozoan and corresponds to the cytosolic form of the enzyme. No introns were detected, and the intergenic noncoding regions do not exceed 50 bp. The mean GC content is close to 60%, and there is a G or C third-base codon bias. Transcription and translation initiation signals also are analyzed, and a model for the mRNA-ssu rRNA interactions is proposed. PMID- 11013708 TI - Mapping chicken genes using preferential amplification of specific alleles. AB - To map the chicken genome, an international reference population was developed at our laboratory (East Lansing, MI) using an F2 backcross between inbred jungle fowl (JF) and inbred white leghorns (WL). To augment the number of type I genes on the East Lansing (E) map, segregation of the JF-specific allele was followed using preferential amplification of specific alleles (PASA) in polymerase chain reactions (PCR). Among 15 functional genes that were added to the E map, agrin and mannose-6-phosphate receptor genes were found to occur in conserved syntenic groups. Using this PCR-based approach, six conserved groups spanning more than 243 centimorgans (cM) in the chicken were syntenic with human and mouse. PMID- 11013709 TI - On the presence and organization of open reading frames of the nonmotile pathogen Brucella abortus similar to class II, III, and IV flagellar genes and to LcrD virulence superfamily. AB - Brucellae are pathogenic, nonmotile bacteria that are facultative intracellular parasites. Little is known about the genetics of these bacteria. Open reading frames from Brucella abortus with similarity to the flagellin, M-ring, and hook of related bacteria were discovered. The open reading frames encode proteins of three of the four flagellum gene classes, namely II, III, and IV. A homolog of the LcrD virulence superfamily was also found. This superfamily is involved in type III protein secretion. B. abortus has the potential for motility and type III secretion. PMID- 11013710 TI - Eggsacting standards. PMID- 11013711 TI - Nothing fishy at FDA seafood laboratory. PMID- 11013712 TI - International round-robin for dioxins and furans. PMID- 11013713 TI - Embalming in the Old Kingdom of Pharaonic Egypt. PMID- 11013714 TI - Exploring electrochemical interfaces with solid-state NMR. PMID- 11013715 TI - Screening and testing for endocrine disrupters. PMID- 11013716 TI - Isolation of amino acids from natural samples using sublimation. AB - Amino acids have appreciable vapor pressures above 150 degrees C and will sublime under partial vacuum at elevated temperatures without any racemization or decomposition. The recoveries of several amino acids including aspartic acid, serine, glycine, alanine, alpha-aminoisobutyric acid, and valine were optimized by varying the temperature and duration of sublimation. Sublimation has been shown to be a rapid and effective technique for the isolation of amino acids from natural samples for enantiomeric analyses and a good substitute for conventional cation-exchange desalting techniques. PMID- 11013717 TI - Voltammetric and pharmacological characterization of dopamine release from single exocytotic events at rat pheochromocytoma (PC12) cells. AB - Although rat pheochromocytoma (PC12) neurotransmitter storage vesicles are known to contain a variety of neurotransmitters including catecholamines, there is little evidence that the molecular species detected during amperometric monitoring of exocytosis is a catecholamine. Rather, as these are catecholamine containing cells, one assumes catecholamines are released. Additionally, although the total amount of transmitter released can be quantified, it has been extremely difficult to evaluate the concentration at the point of release for each exocytosis event. Interpreting voltammograms obtained in the attoliter volume affected between the electrode and the cell and defined by the size of the exocytosis pore during exocytosis is an extreme analytical challenge. Here we use voltammetry of approximately 10(-19) mol released from individual exocytosis events to identify, along with pharmacological evidence, the released compound at PC12 cells as a catecholamine, most likely dopamine. The area of the electrode at which oxidation occurs following an exocytosis event is proportional to the temporal delay prior to acquisition of a voltammogram. This model allows determination of relative concentrations from individual release events and has been used to examine events at control cells and cells incubated with the dopamine precursor, L-3,4-dihydroxyphenylalanine (L-DOPA). Exposure to L-DOPA (100 microM for 1 h) results in 145 detectable events for 11 cells compared to 77 events for 29 control cells, clearly indicating that vesicles can be "loaded" with dopamine. However, the concentrations measured at the electrode surface provide similar distributions for both L-DOPA-treated and control cells. Cyclic voltammetric measurements of relative concentration for zeptomole levels of transmitter in attoliter volumes provide evidence that loading vesicles by increased transmitter synthesis does not lead to elevated concentrations at individual release sites. PMID- 11013718 TI - Analytical applications of cooperative interactions associated with charge transfer in cyanometalate electrodes: analysis of sodium and potassium in human whole blood. AB - Nickel electrodes chemically modified with an interfacial layer of nickel ferrocyanide are shown to be of analytical utility for simultaneously sensing sodium and potassium ions in aqueous solutions, human whole blood serum, and human whole blood. By controlling the charge-transfer characteristics of this versatile interface, interfering blood proteins and potential interferences associated with other alkali cations can be avoided. A solid-state model which explains the excellent simultaneous selectivity and sensitivity of the nickel ferricyanide interface is proposed. PMID- 11013719 TI - Micromethod for the investigation of the interactions between DNA and redox active molecules. AB - A novel microscale and surface-based method for the study of the interactions of DNA with other redox-active molecules using DNA-modified electrodes is described. The method is simple, convenient, reliable, reagent-saving, and applicable for DNA studies, especially those involving microsamples. Information such as binding site size (s, in base pairs), binding constant (K), ratio (K0x/KRed) of the binding constants for the oxidized and reduced forms of a bound species, binding free energy (delta Gb), and interaction mode, including changes in the mode of interaction, and "limiting" ratio K0x0/KRed0 at zero ionic strength can be obtained using only 3-15 micrograms of DNA samples. The method was developed using [Co(Phen)3]3+/2+ (Phen = 1,10-phenanthroline)/double-stranded DNA (dsDNA) modified gold electrodes and [Co(bpy)3]3+/2+ (2,2'-bipyridyl)/dsDNA-modified gold electrodes as model systems. For the [Co(Phen)3]3+/2+/dsDNA-modified gold electrode system, a K2+ of (2.5 +/- 0.3) x 10(5) M-1 and an s of 5 bp were obtained in 5 mM pH 7.1 Tris-HCl buffer solution containing 50 mM NaCl. For [Co(bpy)3]3+/2+/dsDNA-modified gold electrodes, K3+ and s values of (1.3 +/- 0.3) x 10(5) M-1 and 3 bp, respectively, were obtained. While the s values are consistent with those reported in the literature obtained by solution methods, the K values are almost an order of magnitude larger. A transition in the nature of the interaction between dsDNA and [Co(Phen)3]3+/2+, from electrostatic to intercalative with increasing ionic strength, was found in our studies. Negative values of delta E0' for [Co(bpy)3]3+/2+ bound to dsDNA suggest that its interaction with dsDNA is predominantly electrostatic over the ionic strength range of 5-105 mM. The "limiting" ratio K3+0/K2+0 of 22 obtained for [Co(Phen)3]3+/2+ bound to dsDNA at zero ionic strength suggests that electrostatic interactions are predominant over intercalative ones under these limiting conditions. The ratio for [Co(bpy)3]3+/2+ of 16 also indicates that the 3+ form binds to dsDNA more strongly than the 2+ form at zero ionic strength. For [Co(Phen)3]3+/2+/single-stranded DNA (ssDNA)-modified gold electrodes, the nonuniformity of the surface structure of ssDNA-modified gold electrodes greatly complicates the analysis. A system consisting of a dsDNA-modified gold electrode and [Co(tppz)2]3+/2+ (tppz = tetra-2-pyridyl-1,4-pyrazine) was studied by this method, with a K2+ value of (5 +/- 1) x 10(5) M-1 and an 8 value of 7 bp being obtained. PMID- 11013720 TI - Amperometric glucose biosensor based on sol-gel organic-inorganic hybrid material. AB - A new type of sol-gel organic-inorganic hybrid material was developed and used for the production of biosensors. This material is composed of silica sol and a grafting copolymer of poly(vinyl alcohol) with 4-vinylpyridine. It prevents the cracking of conventional sol-gel-derived glasses and eliminates the swelling of the hydrogel. The optimum composition of the hybrid material was first examined, and then glucose oxidase was immobilized in this matrix to demonstrate its application. The characteristics of the biosensor were studied by cyclic voltammetry and chronoamperometry. The biosensor exhibited a series of good properties: high sensitivity (600 nA mmol-1 L-1), short response time (11 s) and remarkable long-term stability in storage (at least 5 months). In addition, the characteristics of the second-generation biosensor with the use of tetrathiafulvalene as a mediator were discussed. PMID- 11013721 TI - Counting of single protein molecules at interfaces and application of this technique in early-stage diagnosis. AB - The fluorescence-based detection and counting of single protein molecules after specific binding to antibodies at interfaces is presented. A diode laser was used as the excitation source. The unspecific binding at the interface has been reduced to a level of only 0.1% of the maximum signal level. At present, the detection limit of this molecule-counting process is in the range of 10(-17) mol/L, and the dynamic range of the signal corresponds to 7 orders of magnitude of antigen concentration, but these values are not limiting. As a preliminary application in early-stage diagnosis, we have investigated the detection of a single cardiac actin molecule in human plasma, which is of interest in myocardial infarction diagnosis. PMID- 11013722 TI - Direct analysis of single rat peritoneal mast cells with laser vaporization/ionization mass spectrometry. AB - A linear time-of-flight mass spectrometer was used as a detector for flow cytometry. These two techniques were coupled by a laser vaporization/ionization interface. The estimated mass detection limit of the combined system was 20 amol of serotonin standard with one laser pulse. An aqueous buffer at physiological pH was used to ensure compatibility with cells. Rat peritoneal mast cells (RPMCs) were dispensed into the mass spectrometer in a single file confined within a 20 micron-i.d. capillary. By using the mass spectrometer as a detector, no precolumn staining or derivatization is required. Determination of serotonin and histamine in individual cells was demonstrated. With this method, hundreds of cells can be analyzed within a few minutes. The average amounts of histamine and serotonin per RPMC were found to be 0.75 +/- 0.33 and 0.11 +/- 0.06 fmol, respectively. No correlation was found between the amounts of the two amines in each cell. PMID- 11013723 TI - Automation of data collection for matrix-assisted laser desorption/ionization mass spectrometry using a correlative analysis algorithm. AB - Automation of data collection in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry using a correlative analysis algorithm is demonstrated. This algorithm was employed to compensate for mass spectral jittering in MALDI data collection (e.g., peak shifts along the m/z axis, signal intensity deviations, etc.). Several important parameters for performing correlative analysis, such as the minimum correlation coefficient to be used and number of mass spectra to acquire prior to correlation, have been investigated and optimized. In addition, the correlation algorithm improved mass resolution of low and high-molecular-weight compounds by as much as a factor of 4. Signal reproducibility in MALDI quantitative analysis also is improved when correlation is employed for data collection. This data collection algorithm can be used in conjunction with other instrumental optimization programs to allow for fully automated MALDI analysis, which is required for the routine applications carried out in many analytical laboratories. PMID- 11013724 TI - Capillary isoelectric focusing-electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for protein characterization. AB - On-line combination of capillary isoelectric focusing (CIEF) with electrospray ionization Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometry is demonstrated for high-resolution analysis of model proteins, human hemoglobin variants, and Escherichia coli proteins. The acquisition of high resolution mass spectra of hemoglobin beta chains allows direct identification of hemoglobin variants A and C, differing in molecular mass by 1 Da. Direct mass determination of cellular proteins separated in the CIEF capillary is achieved using their isotopic envelopes obtained from ESI-FTICR. The factors which dictate overall performance of CIEF-ESI-FTICR, including duty cycle, mass resolution, scan rate, and sensitivity, are discussed in the context of protein variants and cell lysates analyzed in this study. PMID- 11013725 TI - SFE plus C18 lipid cleanup method for selective extraction and GC/MS quantitation of polycyclic aromatic hydrocarbons in biological tissues. AB - Lipid material represents a potential interference for determination of nonpolar compounds (e.g., polycyclic aromatic hydrocarbons) in biological tissue samples. This study reports the development of a selective extraction method using supercritical CO2 that allows the GC/MS quantitation of PAHs in the presence of a substantial lipid background. Selective extraction of PAHs relies upon addition of C18 adsorbent beads to the initial sample slurry. The dried mixture, including C18 adsorbent, is placed in the supercritical fluid extraction (SFE) chamber. During the SFE process, lipids are preferentially retained on the C18 beads. This "SFE plus C18" procedure was developed by first optimizing SFE conditions (100 degrees C, 350 bar) for recovery of PAH standards. PAHs containing added model lipid compounds (stearic acid and cholesterol) were then subjected to SFE plus C18 treatment followed by GC/MS analysis. Using this approach, a recovery of 94 100% of PAHs was obtained while only 9-17% of the lipid material present was coextracted from the same test sample. The developed method is demonstrated to permit efficient recovery and detection of PAHs spiked into crab tissue, a matrix with a high lipid content. PMID- 11013726 TI - Capillary electrochromatography of cannabinoids. AB - The applicability of capillary electrochromatography (CEC) with photodiode array UV detection for the analysis of cannabinoids is presented. Baseline separation of seven cannabinoids (cannabigerol, cannabidiol, cannabinol, delta-9 tetrahydrocannabinol, delta-8-tetrahydrocannabinol, cannabichromene, delta-9 tetrahydrocannabinolic acid) is obtained using a 3-micron CEC Hypersil C18 capillary with an acetonitrile/phosphate (pH 2.57) mobile phase. The effects of acetonitrile concentration, buffer concentration, voltage, temperature, stationary phase, and column length on the separation of the cannabinoids were investigated. Good short- and long-term precision in retention times are observed, with significant improvement obtained using relative retention times with cannabinol as reference compound. Although short- and long-term peak area precisions are poor, satisfactory reproducibility is obtained using relative peak areas with cannabinol as reference compound. The applicability of the CEC methodology to drug seizures was demonstrated on marijuana and hashish. Using a high-sensitivity UV flow cell with an extended path length of 1.2 mm, concentration sensitivities approaching HPLC were obtained. PMID- 11013727 TI - Effect of a predetection open segment in the column on speed and selectivity in capillary electrochromatography. AB - Columns in capillary electrochromatography (CEC) most commonly have the detection window located immediately after the retaining frit of the packed segment. Here, the properties of "duplex" columns having a predetection open segment between the frit and the detector window are examined with particular regard to the effect of the relative lengths of the packed and open segments on the separation of mixtures containing neutral and charged components. This configuration allows the use of columns with short packed segments in contemporary instruments for rapid separations. It is shown that, by varying the length of the packed segment, the balance of chromatographic and electrophoretic forces can be shifted, and the selectivity can be adjusted if the separation involves the interplay of both mechanisms. Expressions are presented for estimating the retention time in a duplex column if the chromatographic and electrophoretic properties of the sample components are known. The results are expected to facilitate CEC method development in selection of the respective column segment lengths for optimum separation. PMID- 11013728 TI - Analysis of nicotine and its oxidation products in nicotine chewing gum by a molecularly imprinted solid-phase extraction. AB - Chromatographic stationary phases showing exceptional selectivity for nicotine can be prepared by the technique of molecular imprinting. Such phases were used in the search for a rapid cleanup step for nicotine and some of its oxidation products in chewing gum formulations. Thus, using an organic mobile phase, the nicotine analytes from chewing gums dissolved in nonpolar solvent were retained, whereas the nonpolar matrix eluted close to the void peak. A subsequent switch to an acidic mobile phase resulted in elution of the analytes as one sharp peak. Due to weak binding of the less basic oxidation products, other imprinted polymers were tested, and the solid-phase extraction procedure was optimized. Polymers were prepared using various functional and cross-linking monomers, templates, porogens and thermal treatments. This resulted in phases that, when compared with a nonimprinted or a C18 reversed-phase column, showed significantly higher recoveries of the analytes. Furthermore, no bleeding of template from the phases could be detected. The cleanup step was coupled off-line to reversed-phase HPLC, and the efficiency of the analysis was compared with and without the cleanup step. Three out of four analytes were quantitatively recovered using the imprinted phase, whereas, using the nonimprinted phase, only nicotine was recovered. Without the cleanup step, none of the analytes could be determined using the reversed-phase HPLC method. PMID- 11013729 TI - Coupled-column liquid chromatography applied to the trace-level determination of triazine herbicides and some of their metabolites in water samples. AB - In the present work, a study is reported of the potential of coupled-column liquid chromatography (LC) applied to the determination of triazine residues in environmental water samples. For this purpose, two different techniques have been compared: on-line trace enrichment followed by LC (SPE-LC) and coupled-column liquid chromatography (LC-LC). First, a completely automated liquid chromatographic method based on on-line trace enrichment in a prepacked precolumn and using diode array detection has been developed for the simultaneous trace level determination of six triazine herbicides (simazine, cyanazine, atrazine, terbumeton, terbuthylazine, and terbutryn) and the main atrazine metabolites (desisopropylatrazine, desethylatrazine, and hydroxyatrazine). After preconcentration parameters were optimized by testing two different sorbents (C18 and PRP-1) in three cartridges with different dimensions, a sample volume of 100 mL was selected in order to achieve maximal solute preconcentration. Detection limits lower than 0.1 microgram.L-1 were obtained even for the most polar analyte (desisopropylatrazine), which presented recoveries of around 30%. The method was validated by means of recovery experiments in groundwater and surface water samples spiked with the analytes at different levels (0.2-2 micrograms.L-1). Afterward, the procedure was successfully applied in a program for monitoring of triazine residues in surface water carried out in a wet area of Castellon, Spain. Different triazine herbicides such as simazine, terbumeton, terbuthylazine, and terbutryn were identified and quantified. The identity of these compounds was confirmed by their absorption UV spectra and by GC/MS analysis. Finally, two rapid, sensitive, and selective procedures, previously developed in our laboratory for the trace-level determination of triazine compounds, both based on LC-LC, were compared with the former procedure. The SPE-LC approach showed a considerable improvement in the global sensitivity at the expense of a decrease in selectivity as well as in sample throughput. PMID- 11013731 TI - Collision-induced dissociation spectra obtained by Fourier transform ion cyclotron resonance mass spectrometry using a 13C,15N-doubly depleted protein. AB - Fourier transform ion cyclotron resonance mass spectra of 13C,15N-doubly depleted cystatin A M65L, produced by Escherichia coli grown on 99.9% [12C]glucose and 99.99% [14N]ammonium sulfate, showed salient monoisotopic peaks composed of 12C and 14N. Collision-induced dissociation spectra were obtained by increasing the capillary-skimmer potential for the electrospray ionization and by extending the trapping time in a radio frequency-only hexapole ion guide. Fragment ions in the spectra could be readily assigned to the amino acid sequence, owing to their markedly improved resolution and sensitivity as compared to those with the natural isotopic composition. Detailed analyses of the fragmentation patterns, facilitated by the use of 13C,15N-doubly depleted proteins, enabled the assignment of approximately 180 fragment ions to the sequence, while natural isotopic cystatin A allowed the assignment of approximately 110 fragment ions. Interestingly, no fragmentation was detected between residues 50-61 and 62-67, which are stretches known to be involved in the antiparallel beta-sheet at the center of the protein. PMID- 11013730 TI - Extraction of aldicarb and its metabolites from excreta and gastrointestinal tissue. AB - Carbamate insecticide screens often include aldicarb and its oxidative metabolites, aldicarb sulfoxide (ASX), and aldicarb sulfone (ASN). The rapid hydrolysis and thermal cleavage of the C-N bond within the carbamate functional group of these compounds produces nitrile transformation products. Nitriles are primary transformation products from aldicarb, its sulfoxide, or its sulfone. However, these nitriles are infrequently monitored. The method reported used acetonitrile/water extraction and HPLC postcolumn derivitization to determine aldicarb, ASX, and ASN from avian excreta and from gastrointestinal (GI) tissue. Recoveries of aldicarb, ASX and ASN from excreta were of 79% +/- 5.4, 120% +/- 7.7, and 93% +/- 6.2, respectively. Recoveries from tissue were 70% +/- 5.0, 80% +/- 12.1, and 85% +/- 6.7, respectively. The same extraction procedure and a GC FPD analysis were used to determine nitrile metabolites from the same tissues. Aldicarb nitrile, ASX nitrile, and ASN nitrile recoveries from excreta were 42% +/- 2.3, 65% +/- 3.6, and 79% +/- 3.3, respectively. Overall recoveries from tissue were 29% +/- 3.4, 72% +/- 8.3, and 83% +/- 11.4, respectively. Since aldicarb, ASX, and ASN are normally detectable in organ tissues for 1-2 days following exposure, determining the presence of nitrile cleavage products provides an important forensic tool for evaluating aldicarb exposures. PMID- 11013732 TI - [Hygienic characteristics of the influence of anthropogenic and natural geochemical factors on the population health in the South Ural]. AB - The natural and technogenic factors of the industrial towns and rural localities in the Orenburg Region were differentially analyzed. The modifying effects of xenobiotics and their complex action on man are considered. The study showed the high pollution of the atmosphere, the blanket of snow, soil, the trace composition imbalance of drinking water, foodstuffs, a reduction in biogenic elements and an increase in toxic elements in the children's substrates (blood, hair) and high morbidity among the children suggest the presence of the formed technogenic biogeochemical province in the Eastern zone and, those in terms of some xenobiotics, in the Central area of the Orenburg Region. PMID- 11013733 TI - [Microbiological analysis of the state of environment in the Orenburg region]. AB - The frequency of the residual staphylococcal carriage among the children who live in rural area having different levels and patterns of environmental pollution is estimated. A relationship is found between the frequency of the residual staphylococcal carriage and the rate of technogenic pollution of the ambient air. The findings suggest that the staphylococcal carriage among children may be used as an index of the unfavorable environment. PMID- 11013734 TI - [Anthropogenic environmental factors and their role in the occurrence of acute respiratory diseases]. AB - The incidence of acute respiratory diseases (ARD) in two districts of the Orenburg Region which have different levels of environmental pollution was comparatively analyzed. The higher incidence of ARD in the more polluted district (Kuvandyksky) than in the control one (Belyaevsky). The incidence rate (41.8%) of ARD correlated with the level of the ambient air pollution by dust, CO, NO2, NF and fluoride aerosols in the Kuvandyksky district. PMID- 11013735 TI - [Hygienic evaluation of staphylococcal nasal mucosal biocenosis in school children of an industrial city]. AB - The results of microbiological spatial monitoring in Orenburg were evaluated. A biological variety of Staphylococcus on the nasal mucosa of schoolchildren were detected. The highest proportion of carriage of the pathogen Staphylococcus was found in different districts of Orenburg, which had the highest air pollution. PMID- 11013736 TI - [Hygienic aspects of air pollution by sulfur-containing compounds]. AB - The physiological and biochemical states of children and adults who resided in the vicinity of the sulphide-containing gas processing plant were studied. The chemical agents emitted by the plant were found to have adverse effects of the children's functional status, namely, decreased vital capacity of the lung, mental performance, retarded sensomotor responses, altered enzymatic system activity. PMID- 11013737 TI - [Hygienic assessment of the effects of drinking water on the population's health]. AB - The paper presents the quality of drinking water. The carcinogenic risk of water and the coefficient of its toxic effect were calculated. The findings determine the priority of the pollutants detected in the drinking water, estimate the carcinogenic risk of water to be 1.4 per 10,000 population. It is recommended to revise specifications for the drinking water of dichloromethane, dichloroethylene, dioctyl phthalate and acrylonitrile. PMID- 11013738 TI - [Organic pollution of the Ural river in the area of open water supply of Orenburg]. AB - The water from the Ural river contains about 230 organic chemical substances, including especially hazardous pollutants. The levels of some chemicals are 2 to 13 times higher than their maximum allowable concentrations. It is necessary to use reliable methods for water treatment at waterworks to lower the levels of drinking water pollutants. PMID- 11013739 TI - [Effect of thermal power stations on the sanitary and biological conditions of water reservoirs]. AB - Discharge of thermal waters from power stations can result in the development of thermophilic microorganisms in the water reservoirs and increased water pollution. Increased water temperature changes relationships between lysozyme active and antilysozyme-active bacteria. The quality of water gets worse. PMID- 11013740 TI - [Hygienic aspects of the use of compact plants for sewage treatment in small localities]. AB - The waste-water system of small localities is an important measure for their sanitary well-being. At present, the firm "Ekobios" produces and installs block module plants for sewage treatment in small localities. PMID- 11013741 TI - [Hygienic characteristics of work conditions during primary metallurgic processing of natural alloy ores]. AB - Natural alloy ores contribute to specific pollution of the industrial environment in the blast-furnace department. The latter is characterized by elevated humidity and SO2, the higher incidence of blood circulatory disorders among the workers. PMID- 11013742 TI - [Role of environmental factors of an industrial city in the etiology of cardiovascular diseases in children]. AB - The prevalence of cardiovascular diseases among 2840 children aged 6 to 15 years and 7000 neonates in two Orenburg districts was studied. High environmental pollution in one of the study districts promotes the high spread of cardiovascular diseases among children. PMID- 11013743 TI - [Preschool children's health in towns with different levels of environmental pollution]. AB - A total of 3497 preschool children residing in the towns having with different man-made burden were examined. The study showed that most children referred to Health Groups 2 and 3. In the town with higher environmental pollution there were a larger proportion of children with dysfunctions of different organs and systems and higher incidence of diseases, including respiratory allergoses, etc. PMID- 11013744 TI - [Regional features of the immune status of schoolchildren in the Orenburg region]. AB - Immunity was studied in 319 schoolchildren from the Orenburg Region. They were found to have lower blood levels of leucocytes and T lymphocytes. It is suggested that imbalance of traces (mainly copper and zinc) is a cause of these changes. PMID- 11013745 TI - [Hygienic assessment of physical and psychophysiologic development of children residing in the area of emissions of Orenburg gas-producing plants]. AB - The children and adolescents residing in rural settlements located in the vicinity of sites of industrial extraction and processing of hydrogen sulphide containing gas were studied. The drinking water and ambient air in these settlements were shown to be polluted. Low indices of the children's mental, physiological, and physical development were revealed. PMID- 11013746 TI - [Biochemical parameters of children residing in an area of high-level technogenic load]. AB - Pollution of the ambient air and drinking water in two comparable regions was investigated. Twenty four metals and traces were found in the children's hair. Higher methemoglobin was found in the blood of the children residing in the higher polluted areas. Decreased concentrations of ascorbic acid were detected in the urine of the same children. It is recommended to develop prophylactic measures to protect children's health. PMID- 11013747 TI - [Radio-ecological and genetic assessment of late consequences of Totsk nuclear explosion]. AB - Radio-ecological and cytogenetic assessments were made in the vicinity of Totsk nuclear explosion in the Orenburg Region. Increased contents of radioactive isotopes Cs and Pu were detected in the soil. A comparative cytogenetic analysis indicated that the population continued to be exposed to radioactive agents. PMID- 11013748 TI - [Health status of children living in rural localities in the area of Totsk nuclear explosion]. AB - The paper presents late outcomes of radiation exposure of 3205 children aged 6-17 years from native persons residing in the areas exposed to radiation 40 years ago. The spread of noncommunicable diseases in the children from the polluted area is 92%. Organic abnormalities are prevalent among them. These include thyroid hyperplasia, lymphadenopathy, gastrointestinal, cardiovascular, and other diseases. Thus, the environmental radiological situation in the southern Urals and the Orenburg Region is poor. PMID- 11013749 TI - [Comparative characteristics of environmental factors of rural localities in the area of Totsk nuclear explosion]. AB - Environmental pollution of rural settlements in the vicinity of Totsk nuclear explosion was investigated. The findings show that the blanket of snow, soil and drinking water in the polluted area did not differ from those in the control district. High levels of stable strontium in the soil and drinking water and those of iodine in the water were detected. The concentrations of plutonium-239, 240 and cesium-137 in the soil were higher than the levels of global sediments and the baseline values of cesium-137 for the Orenburg Region. PMID- 11013750 TI - [Contribution of physical factors to the complex anthropogenic load in an industrial town]. AB - The paper provides evidence for the increasing significance of noise and electromagnetic fields. The correlation coefficients between urban noise and the incidence of digestive diseases were found to be 0.84, those between noise and complication pregnancy were 0.83, those between noise perinatal pathology was 0.79, those between electromagnetic exposure (EME) and neuropsychopathy, EME and cataract, and EME and circulatory disorders were 0.71, 0.91, and 0.78, respectively. PMID- 11013751 TI - [Comprehensive hygienic assessment of anthropogenic pollution of development lands of an industrial town]. AB - Pollution of the ambient air, soil, and drinking water in the Orenburg districts was studied. Comprehensive hygienic assessment revealed high man-made pollution of development lands of the environment, industrial districts of Orenburg town in particular. PMID- 11013752 TI - [Epidemiologic and immunologic parameters in the assessment of endemic goiter in the Orenburg region]. AB - Epidemiological investigations were carried out in 2328 children in the districts of the Orenburg district. Iodine deficiency was detected in the children living in the districts having low concentrations in the water and foodstuffs. There was a deviation of immunological parameters from the normal values in the children of the same districts. PMID- 11013753 TI - [Comprehensive hygienic assessment of environmental pollution in rural localities in the area affected by the Orenburg gas processing plant]. AB - Comprehensive analysis of environmental pollution was made in 15 rural settlements of Orenburgsky and Perevolotsky districts located in the vicinity of gas-processing works. The ambient air, the blanket of snow, and drinking water were assessed for pollution. The drinking water and ambient air in vicinity of the works were found to be polluted by toxic substances. PMID- 11013754 TI - The molecular evolution of Trypanosomatidae. AB - In the absence of a fossil record, theories relating to the evolution of protozoa have, for most of the twentieth century, been based on morphological and life cycle data despite their known limitations. However, recent advances in molecular methodology, notably the wide availability of accurate, automated DNA sequencing, have made it possible to deduce the evolutionary relationships of extant species from their genes. This paper focuses on new findings concerning the evolution of the Trypanosomatidae, based on the ever-expanding body of molecular data now available. Classically, the evolution of digenetic parasitism in kinetoplastids has centred around two opposing theories--invertebrate first or vertebrate first- depending on which was the original host of the monogenetic parasite. However, data supporting a close phylogenetic relationship between genera of monogenetic insect parasites and digenetic vertebrate parasites challenge the simplicity of these hypotheses and suggest that the transition may not have been a major evolutionary barrier. The implications of these observations for the evolution of parasitism within the group are discussed. Phylogenetic analysis of a diverse selection of trypanosomatid species suggests that the genus Trypanosoma is monophyletic and that the human parasites, T. brucei, T. cruzi and Leishmania spp., have fundamentally different patterns of evolution. T. brucei clusters with mammalian trypanosomes of African origin, suggesting an evolutionary history confined to Africa. T. cruzi shows association with trypanosomes from bats, T. rangeli, and trypanosomes from a range of South American mammals and an Australian kangaroo. The origins of most parasites within this clade lie in South America and Australia, suggesting an ancient southern super-continent origin for T. cruzi, possibly in marsupials. The divergence between the Leishmania and Trypanosoma lineages is also ancient. The topology of Leishmania phylogenies suggests an independent transition to digenetic parasitism, a neotropical origin and an early tertiary radiation of the parasite. PMID- 11013755 TI - Transovarial transmission in the microsporidia. AB - The microsporidia are an ancient and diverse group of protists which have many unusual characteristics. These include prokaryotic-like 70s ribosomes, enclosed nuclear division, a lack of mitochondria and complex life cycles which frequently involve vertical transmission. This use of vertical transmission is unparalleled by other protists and is seen only among bacterial endosymbionts and sex ratio distorters and in host cell organelles. Transovarially transmitted microsporidia can have unusual and profound effects on host population sex ratios. We here consider the mechanisms of transovarial transmission and its implications for parasite evolution. We review parasite/host relationships and the evolution of virulence under transovarial transmission and consider the implications of these parasites for host ecology and evolution. PMID- 11013756 TI - Adhesive secretions in the Platyhelminthes. AB - This review is the first to draw together knowledge about bioadhesives secreted by a group of parasites. Mechanisms of mechanical attachment are well known among parasites, but some can also attach to host surfaces by chemical means using a thin layer of adhesive material secreted at the parasite-host interface. Attachment by adhesives to living surfaces has not been studied in detail previously. A significant volume of research has determined much about the chemistry and nature of bioadhesives secreted by various marine macroinvertebrates from different phyla for attachment to inert substrates. Mussels and barnacles are sessile and adhere permanently, whereas starfish display temporary but firm adhesion during locomotion, feeding and burrowing. We focus on the Platyhelminthes that comprises the largely free-living Turbellaria and the wholly parasitic Monogenea, Cestoda, Digenea and Aspidogastrea. The term tissue adhesion is introduced to describe attachment by adhesives to epithelial surfaces such as fish epidermis and the lining of the vertebrate gut. These living layers regenerate rapidly, secrete mucus, are a site for immune activity and are therefore especially hostile environments for organisms that inhabit them, presenting a significant challenge for adhesion. Not all platyhelminths adhere to living surfaces and types of adhesion to inert substrates by the free living turbellarians are also reviewed. Tissue adhesion is particularly well exemplified by monopisthocotylean monogeneans, parasites that are especially mobile as larvae, juveniles and adults on the epidermis of the body and gill surfaces of fish. These monogeneans secrete adhesives from the anterior end when they move from site to site, but some have secondarily developed adhesives at the posterior end to supplement or replace mechanical attachment by hooks and/or by suction. The temporary but tenacious anterior adhesives of monogeneans display remarkable properties of instant attachment to and detachment from their host fish surfaces. In contrast to the mobility of turbellarians and monopisthocotylean monogeneans and the simplicity of their direct life cycles, the largely endoparasitic Cestoda and Digenea are considered to be less mobile as adults. The complex cestode and digenean life cycles, involving intermediate hosts, place different demands on their various stages. Diverse, mostly anterior, gland cells in larvae, metacestodes and adults of the true tapeworms (Eucestoda), and in larval and adult Gyrocotylidea and Amphilinidea are reviewed. Conspicuous gland cells, mostly but not exclusively at the anterior end, in miracidia, cercariae and adults of digeneans and in cotylocidia and adults of aspidogastreans are also reviewed. Unlike turbellarians and monogeneans, accounts of unequivocal adhesive secretions in the Cestoda, but especially in the Digenea and Aspidogastrea, are relatively rare. The primary purpose of many conspicuous glands in the different stages of these mostly endoparasitic flatworms is for penetration into, or escape from, different hosts in their life cycle. We provide a detailed review of current knowledge about adhesion (in the sense of a thin layer of chemical material) in the Platyhelminthes including uses among eggs, larval, juvenile and adult stages. Information on structure, morphology and ultrastructure of the various adhesive systems that have been described is reviewed. Application of the 'duo gland' model is discussed. Comparisons are made between the little that is known about the chemistry of flatworm adhesives and the significant knowledge of the chemical nature of other invertebrate bioadhesives, especially those from marine macroinvertebrates. The potential importance of adhesives in parasitism is discussed. (ABSTRACT TRUNCATED) PMID- 11013757 TI - The use of ultrasound in schistosomiasis. AB - Ultrasound was introduced in the 1970s as a method to detect schistosomal pathology both at hospital and field level. It has since been established as a safe, rapid, non-invasive and relatively inexpensive technique for assessing schistosomiasis-related lesions in individual patients and in community surveys. It can be used to validate laboratory tests to measure morbidity and provides an opportunity to visualize the evolution of pathological lesions after treatment. The interpretation of ultrasound imaging depends on the experience of the investigators and it may not be the ideal tool to detect early lesions of the affected organs. This paper reviews and critically discusses the present knowledge of morbidity due to the different types of schistosomiasis as it can be observed using ultrasound, with special reference to its use as a diagnostic and monitoring tool in field surveys. It analyses the practical use, benefits and drawbacks of ultrasound investigations to assess pathological lesions due to schistosomiasis in relation to other diagnostic tools. The role of ultrasound investigations among other monitoring approaches in control programmes is discussed in the context of rational control strategies. PMID- 11013758 TI - Ascaris and ascariasis. AB - In recent years much new information has been obtained about the epidemiology, population biology and public health significance of infections of Ascaris lumbricoides in humans. Results from experimental infections of A. suum in pigs have helped to elucidate the observations made in the community on human ascariasis. The main purpose of the review is to see how new information may contribute to further acceptance of ascariasis as a serious contributor to ill health and so to the design and implementation of sustainable control programmes intended to reduce the morbidity due to infection with A. lumbricoides. Eradication is neither a realistic nor prudent aim given the current shortage of appropriate sanitation in many countries where ascariasis is endemic. A substantial body of evidence shows that for the four common species of soil transmitted nematode, including A. lumbricoides, regular administration of broad spectrum anthelminthic drugs to children attending primary schools is a cost effective means of controlling the infections. Anthelminthic drugs must be of proven quality and efficacy and health professionals should be prepared to detect and manage drug resistance should that emerge. Despite a deeper understanding of the immune response of a variety of hosts to infections with either A. lumbricoides or A. suum there is at present little prospect of an effective vaccine against ascariasis. The relationship between A. lumbricoides and A. suum is addressed, particularly since both species, if they are indeed separate species, occur in people and their pigs in many communities. PMID- 11013759 TI - HIV therapeutics: past, present, and future. PMID- 11013760 TI - HIV drug resistance and viral fitness. PMID- 11013761 TI - Inhibitors of HIV-1 reverse transcriptase. PMID- 11013762 TI - HIV-1 protease: maturation, enzyme specificity, and drug resistance. PMID- 11013763 TI - HIV-1 integrase inhibitors: past, present, and future. PMID- 11013764 TI - Selection of HIV replication inhibitors: chemistry and biology. PMID- 11013765 TI - Therapies directed against the Rev axis of HIV autoregulation. PMID- 11013766 TI - HIV-1 gene therapy: promise for the future. PMID- 11013767 TI - Assessment of HIV vaccine development: past, present, and future. PMID- 11013768 TI - HIV-1-associated central nervous system dysfunction. AB - Despite more than 15 years of extensive investigative efforts, a complete understanding of the neurological consequences of HIV-1 CNS infection remains elusive. Although the resources of numerous investigators have been focused on studies of HIV-1-associated CNS disease, the complex nature of the disease processes that underlie the clinical, pathological, and cellular manifestations of HIV-1 CNS infection have required a larger volume of studies than was initially envisioned. Several major areas remain as the focus of current research efforts. One of the more pressing issues facing researchers and clinicians alike is the search for correlates to the development of HIV-1-associated CNS neuropathology and the onset of HIVD. Although numerous parameters have been studied, none have been shown to be absolute predictors or markers of HIV-1 related CNS dysfunction. The identification of solid correlates of HIVD is an important goal that would permit clinical identification of individuals at risk for developing potentially crippling, life-threatening CNS abnormalities and would facilitate early treatment of nascent neurological problems. A more complete comprehension of the cellular foundations of CNS dysfunction and HIVD is also a fundamental part of strategies designed to treat or prevent HIV-1 associated CNS disease. Future investigations will strive to expand the body of knowledge concerning the complex interactions between infected and uninfected neuroglial cells and the roles of numerous cytokines, chemokines, and other soluble agents that are deregulated during HIV-1 CNS infection. In particular, a thorough understanding of the mechanisms of neurotoxicity may facilitate the development of new therapies that alleviate or eliminate the clinical consequences of CNS infection. Finally, investigators will continue to study HIVD within the context of single and combination drug therapies used in the treatment of HIV-1 infection and AIDS. As newer and more effective systemic treatments for HIV-1 infection and AIDS are introduced, the effects of these treatments on the onset, incidence, and severity of HIVD will also require intensive study. The impact of drug therapies on the ability of the CNS to act as an HIV-1 reservoir will also need to be addressed. Introduction of each new drug or drug combination will necessitate studies of drug penetration into the CNS and efficacy against the development of CNS abnormalities. Furthermore, as more effective treatments prolong the lifespan of individuals infected with HIV-1, the impact of extended survival on the occurrence and severity of HIVD will also require further investigations. The quest for answers to these and other questions will be complicated by the diversity of experimental systems used to study different aspects of HIV-1 CNS infection and HIVD. Each system has its own unique strengths and weaknesses. Clinical observations provide a continuous spectrum of symptomatic findings but reveal little about the underlying mechanisms of disease. In vivo imaging techniques, such as CT and MRI, also provide a continuum of observations, but the images are limited in their resolution. Neuropathological examinations of postmortem HIV-1-infected brains offer gross, cellular, and molecular views (including phenotypic and genotypic analyses of CNS viral isolates) of the diseased brain, but only provide a snapshot of the end stage neurologic dysfunction. Studies that rely on animal surrogates for HIV-1, including SIV, simian-HIV (SHIV), feline immunodeficiency virus (FIV), visna virus, and HIV-1 SCID-hu models, permit experimental protocols that cannot be carried out in humans, but are limited by the fidelity with which each virus and animal model emulates the conditions and events observed in the human host. Finally, in vitro techniques, which include the use of primary cells and cell lines, adult or fetal human cell cultures, and BBB barrier model systems, are also convenient means by which aspe PMID- 11013769 TI - Molecular mechanisms of human immunodeficiency virus type 1 mother-infant transmission. PMID- 11013770 TI - Molecular epidemiology of HIV-1: an example of Asia. PMID- 11013771 TI - Simian immunodeficiency virus infection of monkeys as a model system for the study of AIDS pathogenesis, treatment, and prevention. AB - As presented in this review, there are a number of different models of both natural and experimental infection of monkeys with primate lentiviruses. There are numerous different viruses and multiple different monkey species, making for a potentially large number of different combinations. The fact that each different combination of virus isolate and host macaque species may show different behavior underscores the need to understand the different models and their key features. On the one hand, this diversity of systems underscores the need to provide some standardization of the systems used for certain kinds of studies, such as vaccine evaluations, in order to facilitate the comparison of results obtained in different experiments, but in essentially the same experimental system. On the other hand, the rich diversity of different systems, with different features and behaviors, represents a tremendous resource, among other things allowing the investigator to select the system that best recapitulates particular aspects of human HIV infection for study in a relevant nonhuman primate model. Such studies have provided, and may be expected to continue to provide, important insights to guide HIV treatment and vaccine development in the future. PMID- 11013772 TI - Animal models for AIDS pathogenesis. PMID- 11013773 TI - [The laryngeal mask in pediatric adenotonsillectomy. A meta-analysis of medical studies]. AB - Anaesthesia both for adenotomy (AT) and for tonsillectomy (TE) frequently presents a challenge. On one hand, children scheduled for adenotomy often have upper airway infections and are thus at risk of laryngo- and bronchospasm; on the other hand the ENT surgeon and the anaesthetist have to share the "workspace" in the patient's mouth. Since the succinyl choline debate in the early 1990s, the question of the best muscle relaxant has gone hand in hand with that of the most appropriate means of securing the airway. The concept of the laryngeal mask as airway was initially greeted with scepticism. Following several years' use of the mask for this purpose in AT and TE in young children, we report our experience and summarise the literature on this topic. The laryngeal mask represents a safe alternative to intubation, provided there is close cooperation with the ENT surgeon. PMID- 11013774 TI - [Dexamethasone for prophylaxis of postoperative nausea and vomiting. A meta analysis of randomized controlled studies]. AB - OBJECTIVE: Randomised controlled trials investigating the efficacy of dexamethasone alone or in combination with other antiemetics to prevent postoperative nausea and vomiting (PONV) were included in a meta-analysis to estimate the relative efficacy of these treatments. METHODS: Studies were systematically searched using Medline, EMBASE, the Cochrane-Library, and by manual screening the reference lists and current issues of locally available anaesthesia journals. Studies identified were divided into four different groups. For each subgroup an independent analysis was performed: 1. Dexamethasone vs. placebo, 2. Dexamethasone + other antiemetic vs. other antiemetic alone, 3. Dexamethasone + other antiemetic vs. dexamethasone alone, 4. Dexamethasone vs. other antiemetics. The main end point in each study was defined as complete absence of nausea, retching, and vomiting after prophylactic antiemetic treatment. The pooled odds-ratios, the relative risk (RR) and the numbers-needed to-treat (NNT) with their corresponding 95%-confidence intervals (given in parentheses) were calculated using a random effects model. RESULTS: A total of 26 studies with 2561 patients were analysed. 1. As a sole antiemetic agent dexamethasone is superior to placebo to prevent PONV (RR: 0.49 (0.15-0.42); NNT: 3.4 (2.5-5.3)). 2. When dexamethasone and an other antiemetic (e.g. a 5-HT3 antagonist) are combined this drug combination is significantly more effective than the single antiemetic without dexamethasone (RR: 0.60 (0.46-0.78); NNT: 7.3 (5.7-10.2)). 3. A similar result was obtained when the dexamethasone combination was compared with dexamethasone alone. The combination is statistically superior (RR: 0.16 (0.08-0.32); NNT: 3.2 (2.2-6.3)). 4. Dexamethasone was usually compared with 5-HT3-antagonist and to a less extends also with dopamine antagonists. Summarising these studies, there was no significant difference concerning effectiveness (RR: 1.35 (0.99-1.85); NNT: 10.6 (5.6-92.6)). CONCLUSION: Dexamethasone has antiemetics effects that are superior to placebo treatment and are comparable with conventional antiemetic agents (e.g. 5-HT3-antagonist, dopamine antagonists). The drug is especially useful in combination with other antiemetics and increases the efficacy of the antiemetic partner drug. PMID- 11013775 TI - [Effectiveness of morphine by periarticular injections after shoulder arthroscopy]. AB - Peripheral opioid receptors have been found in inflamed synovia and the analgesic effect of intra-articularly administered morphine after arthroscopic knee surgery has been proven. There is controversy about efficacy of intraarticular morphine after shoulder arthroscopy. Thirty-two patients with impingement syndrome underwent subacromial decompression in the course of arthroscopic shoulder surgery. At the end of the operation morphine (5 mg) or saline was injected periarticularly. Pain intensity (rest and passive mobilisation) was recorded after recovery and after 1,2,4, and 24 hours (Numeric Rating Scale); the use of rescue medication (piritramide by patient controlled analgesia) was noted. No relevant pain reduction was apparent in the morphine group. Piritramide consumption was identical in both groups (19.7 +/- 16 mg vs. 19.8 +/- 19 mg). We conclude that periarticularly administered morphine in arthroscopic subacromial decompression in the dosage applied in this study does exert no relevant analgesic effect. This is possibly due to the fact that either subacromial tissue, despite of chronic inflammation, does not show the same reagibility as synovia or it is a problem of the nearly complete resection of the subacromial bursa. PMID- 11013776 TI - [Pediatric emergencies. An epidemiologic study of mobile care units in Innsbruck]. AB - OBJECTIVES: We studied the epidemiology and outcome of prehospital pediatric emergencies treated by a physician-staffed mobile intensive care unit (MICU). METHODS: A 3-year retrospective analysis for the period 1991-1993. RESULTS: Children under the age of 15 years comprised 5.1% of the patients treated by the MICU (372/7423), 87.4% of whom were not in a life-threatening condition. The most common emergencies were: trauma (30.4%), febrile seizure (27.7%), and subglottal laryngitis (12.6%). In 44.6% of cases there was no medical indication for the MICU. Intubation at the scene was required by 17 patients (4.6%), and 11 (3.0%) underwent cardiopulmonary resuscitation following prehospital cardiac arrest; two children were successfully resuscitated but died in hospital. A total of 217 (61.3%) were admitted to the ward, 9 of these to a critical care unit. The average length of stay was 4.9 days, and 94.5% of patients were discharged in good health. CONCLUSION: Prehospital pediatric emergencies are rare and seldom life-threatening. Continuing education in pediatric emergency care is important for emergency physicians. PMID- 11013777 TI - [Establishment of a national database for ICU-associated infections. First results from the "Krankenhaus-Infections-Surveillance-System" (KISS)]. AB - OBJECTIVES: To establish a surveillance system as an element of internal quality management, participating intensive care units (ICUs) report their ICU-associated infection surveillance data for aggregation into a national database. METHODS: In order to provide data on ICU-associated infections, a nosocomial surveillance system in German intensive care units (Krankenhaus-Infections-Surveillance-System (KISS)) started in 1997. The method of data collection is based on the (adult) ICU surveillance component from the National Nosocomial Infections Surveillance (NNIS)-System. Until now 113 German ICUs (most of them medical/surgical ICUs) were included in this system. We continuously collected and calculated the data from site-specific infections (device-associated pneumonias, blood stream infections and urinary tract infections). RESULTS: There are now a total of 393,177 patient-days (100,015 patients) among them 176,415 ventilator-days, 295,221 central line-days and 316,799 urinary catheter-days in the data base. The data analysis showed the following device-associated infection rates: 11.2 pneumonias/1000 ventilator-days, 1.8 primary bloodstream infections/1000 central line-days and 4.0 urinary tract infections/1000 urinary catheter-days. CONCLUSION: The project has reached high interest in Germany and animated more ICUs to take part or to apply the same method in order to use the reference data for comparison. PMID- 11013778 TI - [The historical development of intensive care in Germany. Contemporary reflections. 18. Development of hygiene in intensive care]. PMID- 11013779 TI - [ESA 2000]. PMID- 11013780 TI - [Preclinical management of patients with stroke. Remarks and reply to the paper of H.-J. Hennes et al., Anaesthesist 1999) 48: 858-70]. PMID- 11013781 TI - [Perioperative infusion therapy in abdominal surgery]. PMID- 11013782 TI - [Tourniquets and muscle relaxation]. PMID- 11013783 TI - [Pulmonary gas exchange in anesthesia]. PMID- 11013784 TI - Prostate cancer. What men want from their family physicians. PMID- 11013785 TI - Privileging and consultation in maternity and newborn care. Maternity and Newborn Care Committee. PMID- 11013786 TI - Concerns about the critical appraisal of the role of H pylori in dyspepsia. PMID- 11013787 TI - Be careful about drawing conclusions. PMID- 11013788 TI - Don't shoot the messenger. PMID- 11013789 TI - Tips from a retired family physician. PMID- 11013790 TI - Canadian Family Physician: through a glass darkly. PMID- 11013791 TI - Which drugs are contraindicated during breastfeeding? Practice guidelines. AB - QUESTION: Many breastfeeding mothers are concerned about taking medications that might affect their babies. Are there any guidelines on which drugs are safe? ANSWER: Only a few drugs pose a clinically significant risk to breastfed babies. In general, antineoplastics, drugs of abuse, some anticonvulsants, ergot alkaloids, and radiopharmaceuticals should not be taken, and levels of amiodarone, cyclosporine, and lithium should be monitored. PMID- 11013792 TI - Ophthaproblem. Peripheral corneal thinning secondary to rheumatoid arthritis. PMID- 11013793 TI - Radial head fracture. PMID- 11013794 TI - Restless legs syndrome. PMID- 11013795 TI - Short report: what do men with prostate cancer want to know? PMID- 11013796 TI - [International Prostate Symptom Scale. Evaluation of the usefulness of a French version]. AB - OBJECTIVE: To evaluate the usefulness of a French-language version of the International Prostate Symptom Scale (I-PSS) by measuring, on this scale and on the quality of life index, the scores of patients with benign prostatic hyperplasia before and after prostate surgery. METHOD: The questionnaire was completed by 14 men, mostly between 60 and 80 years old, 24 hours before surgery and 1 month and 3 months after surgery. RESULTS: The French-language scale worked well. Scores changed from 19.1 before surgery to 7.5 3 months after surgery for prostate symptoms and from 8.5 to 4.5 for quality of life. CONCLUSION: This version of the questionnaire is a valid tool for evaluating prostate symptoms reported by French-speaking people. PMID- 11013797 TI - Referral of children with otitis media. Do family physicians and pediatricians agree? AB - OBJECTIVE: To determine factors influencing family physicians' and pediatricians' decisions to refer children with recurrent acute otitis media (RAOM) and otitis media with effusion (OME) to otolaryngologists for an opinion about tympanostomy tube insertion. DESIGN: Mailed survey. SETTING: Physicians' practices in Ontario. PARTICIPANTS: Random sample of 1459 family physicians and all 775 pediatricians in the province. MAIN OUTCOME MEASURES: Physicians' reports of the influence of 17 factors on decisions to refer (more likely, no influence, less likely to refer) and number of episodes of otitis media, months with effusion, level of hearing loss, or months of continuous antibiotics without improvement prompting referral. RESULTS: Physicians agreed (> 80% concordance) on six out of 17 factors as indications for referring children with RAOM or OME. Opinions about the importance of other factors varied widely. Family physicians would refer children with otitis media after fewer episodes of illness, fewer months of effusion, lower levels of hearing loss, and fewer months of prophylactic antibiotic therapy than pediatricians (all P < .001). Pediatricians would prescribe continuous antibiotics longer (11.8 weeks) than family physicians (8.9 weeks, P < .0001), which correlated with lower referral thresholds for family physicians. CONCLUSION: Family physicians' and pediatricians' self-reported referral practices for surgical opinions on children with otitis media varied considerably. These observations raise questions about the consistency of care for children with otitis media and whether revised clinical guidelines would be helpful. PMID- 11013798 TI - Is respite care available for chronically ill seniors? AB - OBJECTIVE: To determine family physicians' perceptions of how available respite care is and how easy it is to refer chronically ill older people to it, and to examine their opinions of respite care. DESIGN: Mailed survey to family physicians on the Thames Valley Family Practice Research Unit's mailing list. SETTING: London, Ont, and surrounding area. PARTICIPANTS: Of the 448 surveys mailed to eligible physicians, 288 were completed and returned for a response rate of 64.3%. MAIN OUTCOME MEASURES: Respondents' perceptions of how available respite care is and how easy it is to refer chronically ill older people to it and their opinions on the effectiveness of respite care. RESULTS: More than half the respondents reported that outpatient respite care is always available, but how available depended on practice location. Inpatient respite care was reported as less available. More than half the respondents found referral to respite care difficult. Respondents were very positive about the role of respite services in long-term care and in lowering caregiver stress. Respondents' perceptions varied according to where they had attended medical school. Their perceptions of respite care's role in long-term care and in helping patients remain at home were influenced by whether they thought respite care was available. CONCLUSION: Family physicians need education in the value of respite services for their chronically ill older patients and their families. Physicians also need information on the respite services available and strategies for accessing them. Our findings suggest a need for greater attention to regional discrepancies in availability of services. PMID- 11013799 TI - Treating obesity. Lost cause or new opportunity? AB - OBJECTIVE: To review therapies for treating obese patients. QUALITY OF EVIDENCE: Advice in this paper is based mainly on the results of randomized controlled trials. Some data from smaller, more physiologic studies are included. When appropriate, advice is based on consensus. MAIN MESSAGE: Recent medical evidence indicates that a modest weight loss (5% to 10%) can alleviate symptoms of obesity related comorbidity. Treatment of obesity should be comprehensive and integrated into a multi-component approach and should involve both patients and their families. The main challenge of obesity is maintaining a reduced weight. CONCLUSION: A multi-component approach to treating obesity can help make treatment less frustrating and more rewarding for patients and physicians. PMID- 11013800 TI - Status epilepticus. Current concepts and management. AB - OBJECTIVE: To inform primary care physicians about current issues around generalized convulsive status epilepticus (GCSE) emphasizing definition, pathophysiology, treatment, and prognosis. QUALITY OF EVIDENCE: MEDLINE (1994 to 1999) provided 479 references using the MeSH terms "status epilepticus" and "treatment." From these we selected 30 English-language articles covering clinical aspects, treatment, and animal research. Key source documents from previous years and information from modern textbooks and recent symposia were also included. MAIN MESSAGE: Generalized convulsive status epilepticus continues to be a medical emergency with high morbidity and mortality. It must be managed promptly and effectively. The operational definition of GCSE is a seizure that lasts longer than 5 minutes or two or more seizures between which patients do not recover. Main differential diagnosis is nonepileptic status. Intravenous therapy with combined lorazepam and phenytoin is the initial treatment of choice. Other preferred medications are diazepam, midazolam, and propofol. Some of these medications should be considered before arrival at hospital. Prognosis of GCSE is determined by underlying cause, delay in adequate treatment, and comorbidity. Patients with GCSE lasting longer than 30 minutes require intensive care and electroencephalogram monitoring. CONCLUSION: Intravenous lorazepam and phenytoin are currently the most effective drugs for initial management of GCSE. Timely administration of antiepileptic medication can prevent development of GCSE in some patients with known epilepsy. Main differential diagnosis is nonepileptic status. PMID- 11013802 TI - Canadians' smoking behaviour. Results from the National Population Health Survey. Centre for Chronic Disease Prevention and Control. PMID- 11013801 TI - Scaphoid fracture. Review of diagnostic tests and treatment. AB - OBJECTIVE: To help make diagnosis and treatment of scaphoid fracture more precise by review of published evidence. QUALITY OF EVIDENCE: MEDLINE was searched using the terms "scaphoid," "carpal navicular," "fracture," "computed tomography," "bone scan," and "scintigraphy." Most papers were case-series observational reports. Papers were cited if the case series was large or if there was a high degree of agreement among several observers. The main recommendation for change in treatment of scaphoid fracture is based on two randomized clinical trials involving more than 1000 patients with proven scaphoid fracture. MAIN MESSAGE: Fracture of the scaphoid requires a specific mechanism of injury. "Snuffbox" tenderness is not specific for scaphoid fracture and is not the most useful physical finding; other physical findings provide more specific evidence for or against scaphoid fracture. Physical examination remains the basis of initial treatment and should be thorough and meticulous. X-ray films must be of high quality and should be examined carefully for bone and soft tissue signs of fracture. A Colles'-type short arm cast is adequate for treating common undisplaced scaphoid waist fractures; the thumb need not be immobilized. For suspected scaphoid fractures, without radiologic evidence of fracture, treating symptoms is likely sufficient. CONCLUSION: Evidence found in the literature can be used to improve diagnostic accuracy for scaphoid fractures, to optimize treatment for these injuries, and to reduce unnecessary immobilization and disability for patients. PMID- 11013803 TI - Doctor, heal thyself. PMID- 11013804 TI - Lessons for Canada. The 13th International AIDS Conference. PMID- 11013805 TI - [Surgical and multimodal therapy of pseudomyxoma peritonei]. AB - The rarity of Pseudomyxoma peritonei, its complex biology and the remaining inconsistencies in terminology all contribute to rendering therapeutic recommendations a difficult task. In principle, a multimodal concept combining cytoreductive surgery with peri- and postoperative intraperitoneal chemotherapy constitutes the preferable treatment. Initially, surgical peritonectomy aims to achieve the complete removal of tumor cells at the macroscopic level. Subsequently in the early postoperative phase when the absence of adhesions allows for a homogeneous intraabdominal spread of cytotoxic drugs, intraperitoneal application of 5-fluorouracil and mitomycin C represents the most solidly established treatment. Therapeutic failures are often due to insufficiently radical cytoreductive surgery or a lack of homogeneous distribution of cytotoxic drugs in the abdomen. In view of the rarity of their condition, patients with Pseudomyxoma peritonei should be treated at or in conjunction with specialized centers. Only such a "center-oriented" approach will secure the standardization of treatment and help to clarify unsettled therapeutic questions. The development of improved therapeutic concepts will depend on concise histopathological classification. Already at present therapeutic decisions should not only be be based on the clinical aspect of a "mucinous abdomen" but should be guided by the pathological differentiation between disseminated peritoneal adenomucinosis (DPAM), with a relatively good prognosis, and the more aggressive peritoneal mucinous carcinomatosis (PMCA). PMID- 11013806 TI - [Malignant mesothelioma of the pleura, pericardium and peritoneum. 1: Etiology, pathogenesis, pathology]. AB - The incidence of primary malignant neoplasms of the pleura, the pericardium and the peritoneum in Germany has been rising since about the mid-1980s. A continuing rise is expected until about 2020, predominantly due to the peak of asbestos processing in Germany between 1965 and 1980. About 90% of the mesotheliomas stored in the files of the German Mesothelioma Registry in Bochum are asbestos related and therefore possibly due to occupational exposure that may be compensated by the professional associations. More than 500 mesotheliomas annually can be diagnosed in the German Mesothelioma Registry. In this first part of the series on mesotheliomas, current concepts on etiology and pathogenesis as well as diagnostic procedures and standards are discussed. At the present, specific chromosomal or genetic defects that constantly give rise to a mesothelioma are not known. The initiation and progression of malignant mesotheliomas is a highly complex mechanism that is based on individual genetic alterations. A reliable diagnosis is the basis for therapeutic, prognostic and medicolegal consequences; in general, it can be achieved by thoracoscopic inspection with selected biopsy. Surgery may gain a more important role in the therapy of malignant tumors of serosal membranes not only in palliative, but also in potential curative approaches if the diagnosis can be made at earlier tumor stages. PMID- 11013807 TI - [Mesothelioma of the pleura and peritoneum. Diagnostic and therapeutic sequelae]. AB - In patients with pleural or peritoneal mesothelioma, surgery is a technically difficult procedure. Whereas those rare forms of localized pleural mesotheliomy are being detected incidentally and can be cured by complete resection, most patients with diffuse malignant mesothelioma present with an advanced stage of disease. Most of these patients survive less than 12 months irrespective of the treatment modality. For diffuse pleural mesothelioma, some favorable prognostic factors were identified: IMIG (International Mesothelioma Interest Group), stages I and II, epithelial type, age under 50, female gender. In IMIG stages I and II, extended pleuropneumonectomy followed by chemo- and/or radiotherapy is recommended. For this subset of patients, a median survival time of between 20 and 30 months is reported. Pleurectomy and decortication are recommended as palliative surgical strategies against pleural effusion. In patients with technically inoperable infiltration of the thoracic wall, irradiation is helpful; sometimes partial remission and relief of pain can be achieved by chemotherapy. PMID- 11013808 TI - [The desmoid problem]. AB - Desmoid tumors (DTs) are rare nonmetastasizing neoplasms, occurring both sporadically and in the context of familial adenomatous polyposis (FAP). DTs show an unpredictable natural course with approximately 10% demonstrating a severely aggressive growth pattern, whereas others regress spontaneously. Surgery appears to have a triggering effect on desmoid growth, but is unavoidable in patients with FAP requiring prophylactic colectomy. Treatment recommendations remain controversial. Surgery may only be regarded as an option if a safety margin of at least 2 cm is guaranteed. Even under these circumstances recurrence rates are very high. Debulking operations invariably lead to a boost towards more aggressive growth. Most issues regarding DTs are contradictory, reflecting the lack of any systematic approach to the disease. One way out of this dilemma is the establishment of a clinical classification as a first step towards standardized therapy. PMID- 11013809 TI - [Extra-abdominal fibromatosis--extra-abdominal desmoid. Review and personal experiences]. AB - Extraabdominal desmoids represent one group of deep fibromatoses. These aggressive nonmetastasizing tumorlike lesions have a strong tendency to local infiltration, with a recurrence rate of about 40%. Trauma, hormones and heredity have been implicated as etiologic factors. Shoulder, chest wall, back and thigh are favored sites. By combination of different diagnostic procedures the number of differential diagnoses can be reduced to only a few. While in former times surgery was thought to be the only kind of therapy, nowadays adjuvant procedures like radiation, hormonal therapy and also chemotherapy are becoming more and more important. Amputation or other mutilating procedures should be done only if the tumor recurs repeatedly. PMID- 11013810 TI - [Evidence-based recommendations for perioperative antibiotic prophylaxis]. AB - The aim of perioperative antibiotic prophylaxis (PAP) is to reduce postoperative complications. In 1994 specialist American companies developed a PAP quality standard, whereby individual recommendations were divided into categories according to the degree of evidence. These recommendations have been tabulated and supplemented with the results of new, randomized, controlled studies. PMID- 11013811 TI - [Abdominal compartment syndrome: prevention and treatment]. AB - Abdominal compartment syndrome is defined by increased intraabdominal pressure above 20 mmHg with increased pulmonary peak pressure and oliguria. In primary abdominal compartment syndrome the increased intraabdominal pressure is caused directly by peritonitis, ileus or abdominal and pelvic trauma. Secondary compartment syndrome is a result of forced closure of the abdominal wall after abdominal surgery. The effects are decreased cardiac output, pulmonary atelectasis, oliguria to anuria and hepatic as well as intestinal reduction of perfusion. Effective monitoring is done by standardised measuring of urinary bladder pressure. Normal values are between 0 and 7 cm H2O, after elective laparotomies 5-12 cm H(2)0. Above 25 cm H(2)0 they are definitely pathological. For the prevention and therapy of manifested abdominal compartment syndrome the application of a laparostomy using a resorbable mesh is recommended. Between 1988 and 1999 we applied a laparostomy to lower the intraabdominal pressure in 377 patients. In 16% of the cases it was indicated by primary abdominal compartment syndrome with a bladder pressure of 31 +/- 4 cm H(2)0 preoperatively, which could be lowered to 17 +/- 4 cm H(2)0 by laparostomy. An early reconstruction of the abdominal wall could be performed in 18% of the cases. CONCLUSIONS: The abdominal compartment syndrome is an often underestimated problem in abdominal surgery involving multiple organ systems. The temporary laparostomy lowering intraabdominal pressure rather than a forced closure of the abdominal wall should be used in all circumstances. PMID- 11013812 TI - [Idiopathic megacolon. New findings on histopathology and musculo-mechanical causes]. AB - Nineteen native surgical specimens of an idiopathic megacolon were histopathologically investigated and a lack of the tendon-like collagen III movement net of circular and longitudinal muscles observed, including a lack of the connective tissue layer of myenteric plexus. This disease of the muscularis propria could be considered as a desmolysis, a hypoplastic or an aplastic desmosis. With reference to the normal muscle mechanics of the gut, it is shown that the pathogenetic principle of the idiopathic megacolon consists in absence of the tendon-like connective tissue net in the muscularis propria which results in a lack of peristalsis of the gut, despite of a regular enteric innervation. This finding demonstrates that an idiopathic megacolon can be histopathologically verified in whole-mount biopsies and referred for curative surgical treatment. PMID- 11013813 TI - [Can continence function after rectal resection be prognostically estimated?]. AB - AIM: To determine clinical and physiologic parameters enabling the prognosis of continence after protective ileostomy closure secondary to rectal resection for rectal cancer. METHODS: Patients who had undergone rectal resection (n = 65, of whom 24 had had radiochemotherapy) were evaluated by clinical examination, anorectal manometry and orthograde contrast enema before ileostomy closure. Continence was evaluated by clinical findings 91 +/- 52 weeks after stoma closure with the help of standardized questionnaires and classified according to the Wexner continence score. The relationship between findings before stoma closure and continence score was calculated with Pearson's correlation coefficient. RESULTS: Correlations were found to be significant between the continence score and the level of anastomosis (r = -0.58, p < 0.001), median resting pressure (r = -0.52, p < 0.001), rectal compliance (r = -0.43, p < 0.001). Additionally, radiochemotherapy impairs continence (p = 0.0001). Correlations were not significant between continence and functional sphincter length, squeeze pressure, threshold for perception, urge and maximal tolerable volume, and continence for semiliquid contrast medium. CONCLUSION: Incontinence after rectum resection is multifactorial: the level of anastomosis, resting pressure, rectal compliance and radiochemotherapy all play a dominant role. Based on these findings, the continence score can be calculated before closure of a diverting ileostomy by applying multivariate analysis with the help of the following formula: Continence score = 18.23 - 0.94 x level of anastomosis - 0.18 x resting pressure + 3.72 x radiochemotherapy. PMID- 11013814 TI - [Laparoscopic hernia therapy (TAPP) as a teaching operation]. AB - INTRODUCTION: Because of an increasing number of endoscopic hernia procedures, it is important to look into the possibility of standardizing these techniques helping surgeons to acquire the operative skills necessary. MATERIAL AND METHODS: To discuss these aspects, the documented data on TAPP operations that have been carried out in this department since 1993 were analyzed. The results of teaching procedures were compared with those of experts after they had gotten past the learning and development curve. RESULTS: A total of 778 teaching procedures were performed by 10 surgeons with an individual experience of 30.5 operations (median). Before starting the first procedure, 89 were done by assistants operating the camera. The morbidity of teaching operations was 1.9% compared to 1.4% for those performed by experts. After a median follow-up of 23 months there were two recurrences (0.23%) in the expert group and none in the teaching group (follow-up 16 months). CONCLUSION: Because of the potentials of standardization of the TAPP technique, the results of teaching were equal to expert operations. Therefore, TAPP is suitable for application in a routine setting. PMID- 11013816 TI - [Quality of life and subjective evaluation of outcome quality 3 years after laparoscopic antireflux surgery]. AB - PURPOSE: Evaluation of quality of life data and patient satisfaction to estimate the outcome of laparoscopic antireflux surgery (LARS) is nowadays an important issue, the long-term outcome of this has not yet received much attention. METHODS: In the present study we evaluated the outcome of quality of life data of 70 patients who underwent "floppy" Nissen fundoplication at our institute 3 years after surgery. Quality of life was evaluated with the Gastrointestinal Quality of Life Index (GIQLI). Additionally the subjectivity and objectivity of the quality of the procedure and possible side effects were evaluated with a questionnaire. RESULTS: Three years after laparoscopic Nissen fundoplication, patients gave their quality of life (GIQLI) in an overall score of 123.9 points. This is comparable to 122.6 points in the normal population. There was no difference detectable in the subdimensions of GIQLI. Ninety-eight percent of the patients estimated their satisfaction with the procedure as excellent or good and would undergo surgery again if necessary. Four patients suffered from minimal side effects from the procedure, but had no decrease in their quality of life. None of the patients needed antireflux medications postoperatively. Laparoscopic redo fundoplication was performed in two patients 3 months after initial surgery because of persisting dysphagia. CONCLUSION: The efficacy and long-term outcome of treatment of gastroesophageal reflux disease with laparoscopic "floppy" Nissen fundoplication can be evaluated by objective testing, but also by subjective judgment of the patient and with an evaluation of quality of life. PMID- 11013815 TI - [Effects of strength- and endurance oriented training during regional chemotherapy in metastatic rectal carcinoma. Case report as a contribution to surgical oncology]. AB - OBJECTIVE: How does adjuvant training of strength and endurance influence the muscular, cardial, respiratory, immunological systems, and the quality of life during intrahepatical chemotherapy in a veteran athlete with liver metastasis after resection of a carcinoma of the rectum. PATIENT AND METHOD: A 58-year-old athlete with metastatic carcinoma of the rectum (pT3, N0, M-liver, G2) underwent regional chemotherapy (folinic acid, 5-fluorouracil) via an intrahepatic port system. Six cycles with 5 days each were applied. During the intervals between cycles (14 days), beginning in the 6th postoperative week, a strength and endurance training (duration 13 weeks) was carried out twice weekly with an intensity of 40-60% of the maximum postoperative individual power and endurance (evaluated with weight and treadmill exercise tests and measurement of lactate). Before and after chemotherapy, an echocardiogram, resting and exercise electrocardiogram, lung function, natural killer cells and the gastrointestinal quality of life index (GLQI) was evaluated. After chemotherapy was finished, the effects of training were assumed. RESULTS: Strength increased between 0% and 144%. Endurance improvement was measured on the last comparable submaximal intensity step, expressed by reduction of heart rate and lactate concentration by 10 and 21.5%, respectively, improvement of lung function regarding FEV 1 and FCV by 12.9 and 11.3%, respectively, and VC IN 11.4%. The relative count of natural killer cells increased by 27.2%, and the GLQI improved from 109 points (pathologic) to 129 points (normal). CONCLUSION: There were positive effects on muscular and cardial mechanisms of adaptation and on illness-related quality of life. Postoperative sports activity during regional chemotherapy is possible. PMID- 11013817 TI - [Tolerance of early oral feeding after operations of the lower gastrointestinal tract]. AB - INTRODUCTION: Oral feeding is usually offered following surgery of the lower gastrointestinal tract when clinical signs of normal intestinal motility are present. However, some studies have shown that early oral feeding is well tolerated with low morbidity. METHODS: A prospective cohort study was performed to evaluate whether early oral feeding according to a standardized schedule is tolerated under normal clinical circumstances. One hundred consecutive patients following small- or large-bowel resection with anastomosis were offered fluids on post-operative day 1, soup on post-operative day 2, mashed food on post-operative day 3 and a regular diet on post-operative day 4. Parenteral nutrition was only given if necessary. Tolerance of oral feeding and the amount of food were checked twice a day. End points of the study were nausea (VAS score 1-100), vomiting (> 200 ml), reinsertion of a nasogastric tube, level of food intake, parenteral nutrition (ml), appetite and well-being. RESULTS: Loop ileostomies were done in 21 patients, colonic resections above the sigmoid in 32, and sigmoid and rectal resections in 47. The average age was 63 +/- 13 years. The frequency of nausea was less than 30% and of vomiting less than 10%. Only in two cases was a nasogastric tube inserted. Forty-three percent of all patients tolerated feeding very well according to the schedule. On post-operative day 3 more than 60% tolerated oral intake, on post-operative day 4, 74% and on post-operative day 5, 88%. Only 22% of the patients needed parenteral fluids on post-operative day 4. The first bowel movement was noted after 2.8 +/- 1.1 days. Surgical complications were documented in 18 patients and general complications in 6 patients. CONCLUSION: Most patients tolerated early oral feeding very well according to the schedule with low morbidity. Therefore, early feeding is now a substantial component of the postoperative treatment following small- or large-bowel resections. PMID- 11013818 TI - [Development of obstructive jaundice after hemorrhage into a benign, non parasitic liver cyst]. AB - Benign, non-parasitic liver cysts are usually asymptomatic and found incidentally by abdominal ultrasound or CT scan. We present the case of a 68-year-old patient who showed obstructive jaundice due to a hemorrhagic liver cyst compressing the choledochal duct. Due to the location of the cyst in the porta hepatis with partial compression of central liver vessels and the meanwhile organized hematoma, operative therapy was preferred in order to prevent secondary complications such as portal vein thrombosis and to exclude a malignant neoplasm. PMID- 11013819 TI - [Paratesticular sarcomas. Case report of leiomyosarcoma]. AB - Paratesticular sarcomas are rare. We discuss this entity with the aid of a case report and the existing literature. The therapy of these tumors includes resection and possible adjuvant radiotherapy. Rhabdomyosarcomas are an exception, because they also show a good response to chemotherapy. PMID- 11013820 TI - [Surgical principles in stomach carcinoma]. PMID- 11013821 TI - [Does history still have current applications?]. PMID- 11013822 TI - [Patient information for and about the hospital]. PMID- 11013823 TI - [Occupational evaluation--assessment of the most recent law regarding occupational jurisprudence on form and content]. PMID- 11013824 TI - Effect of blocking alpha-adrenoreceptors on the noradrenaline-induced changes in mouse uterine motility during different terms of pregnancy: in situ experiments. PMID- 11013825 TI - Paradoxical sleep considerably contributes to the night decrease in brain temperature in hibernating mammals in summer. PMID- 11013826 TI - Seasonal changes in the effect of L-DOPA and 5-OTP on the integrative activity of suslik brain. PMID- 11013828 TI - Adrenergic regulation of the circulating pool of phagocytic cells in acute stress. PMID- 11013827 TI - Reinforcement effect of glucocorticoid hormones. PMID- 11013829 TI - The effect of 6-hydroxydopamine on elicitation of long-term sensitization in the edible snail: electrophysiological study. PMID- 11013830 TI - Expression of alpha 2-adrenoreceptors in the brain cortex depends on the noradrenaline concentration. PMID- 11013831 TI - Synthetic pineal peptide inhibits growth of spontaneous tumors and increases lifespan in mice. PMID- 11013832 TI - Critical bandwidths of different types of neurons in the mouse auditory midbrain. PMID- 11013833 TI - Effect of short-term pulse adaptation to hypobaric hypoxia on epileptiform seizures and rheological blood properties in KM rats. PMID- 11013834 TI - A pineal peptide increases lifespan in Drosophila melanogaster. PMID- 11013835 TI - DNA damage and tumor cell death caused by the combined effect of vitamins B12b and C. PMID- 11013837 TI - Contribution of induced autolysis to digestion in secondary consumers (examplified by hydrobionts). PMID- 11013836 TI - Studies of synthetic attractants of the Siberian moth Dendrolimus superans Butl. (Lepidoptera: Lasiocampidae). PMID- 11013838 TI - Dwarf males and reproductive tactics in the mikizha Parasalmo mykiss Walbaum (Salmonidae, Salmoniformes) from the Kamchatka peninsula. PMID- 11013839 TI - Ocular rectus muscles in the hypophyseal fossa of the basisphenoid in the permian theriodont Suchogorgon golubevi (Reptilia). PMID- 11013840 TI - Do mammalian karyotypes evolve towards equalization of relative chromosome lengths? PMID- 11013841 TI - Disruption of the dynamics of food passage through the digestive tract in red voles (Clethrionomys rutilus) dwelling in an area near oil and gas extraction. PMID- 11013842 TI - Eye structure in arrow worms (Chaetognatha). PMID- 11013843 TI - Biocatalysis of matter transfer in a microcosm is inhibited by a contaminant: effects of a surfactant on Limnea stagnalis. PMID- 11013844 TI - Unusual hair structure in sloths (Edentata: Bradypodidae). PMID- 11013845 TI - Pedomorphosis in fossil and modern testudinata (exemplified by Trionychidae). PMID- 11013846 TI - The insect as a consumer: a model of efficient behavior. PMID- 11013847 TI - Morphogenetic response of the water vole (Arvicola terrestris L.) to atypical environmental conditions. PMID- 11013848 TI - Immunogenetic characteristics of patients with stomach cancer and predominant activity of either sympathetic or parasympathetic autonomic nervous system. PMID- 11013850 TI - Conodont (Euconodontophyles), a living fossil. PMID- 11013849 TI - Effect of the agouti gene on the hypothalamic-hypophyseal-adrenal system in mice. PMID- 11013851 TI - A new class of mutations in Drosophila melanogaster. PMID- 11013852 TI - A simple model of the spatial dynamics of the consumer-resource system. PMID- 11013853 TI - Induction of potassium transport in wheat sprout roots by low potassium content in the medium. PMID- 11013854 TI - Suppression of symbiotic digestion in red voles (Clethrionomys rutilus) as a result of anthropogenic impact. PMID- 11013855 TI - Protein synthesis inhibitors applied for a short time cause resumption of the resting state of NIH 3T3 strain cells more effectively than growth factors. PMID- 11013856 TI - Effect of ATP on different Ca2+ channels in PC12 pheochromocytoma cells. PMID- 11013857 TI - [Serum tyrosine in children with phenylketonuria and mild hyperphenylalaninemia]. AB - Serum tyrosine concentration, Phe/Tyr scores and psychomotor/mental development scores were analysed in 32 children with phenylketonuria (PKU) and 39 with mild hyperphenylalaninaemia. Observation period included the first 6 years of life. Tendency to tyrosine deficiency was observed; stronger in dietary treated PKU patients than in those with mild hyperphenylalaninaemia. Statistically significant differences between patient groups were found only in 3 and 6 years old children (lower tyrosine values in PKU patients). It was observed that evaluation of Phe/Tyr score value might be usefull in differentiation between PKU and mild hyperphenylalaninaemia. Moreover, the above score may help in the evaluation of hypo- and hyperalimentation state in the course of dietary treatment. The level of tyrosine deficiency in the analysed patient groups did not influence their normal intellectual development. PMID- 11013858 TI - [Sudden infant death syndrome (SIDS). Prevention recommendations]. AB - Etiology of sudden infant death syndrome still remains unknown. However, the risk factors are established and serve as a base for creating the recommendations for infants' sleep safety and SIDS prevention. PMID- 11013859 TI - [Apnea states in infants, anxiety of parents and pediatricians, and home monitoring of respiration in children]. AB - In order to determine parental reasons of the use of home apnea/respiration monitors and to estimate the grounds of these decisions correspondence contact was achieved with the parents of 71 infants monitored by means of the APNEAL-1 device. Filled up questionnaires were the source of data regarding the families (structure, health practices, mothers' age and education, course of pregnancy and delivery), monitored infants (birth weight, Apgar score, apnea events) and the reasons of monitoring. Also opinions concerning monitor usefulness and performance were gathered. The majority of our families (> 97%) consisted of both parents, their socio-economic status was medium (52.11%) or good (46.48%) and they were residents of cities (88.73%). Cigarette smoking was present in 22 families (31%) with 13 smoking mothers (18.31%). Mothers' mean age was 29.11 years (SD 4.84), 45% of them had high school education and 36.62%-university education. As many as 45% of the pregnancies had a pathological course and 34.3% were terminated by a preterm delivery. Mean birth weight of monitored children was 2914.93 g (SD 971.4), mean Apgar score was 8.31 (SD 2.46). More than 60% of children were breast fed during the first six months of life, and 30% of them- during first year of life. Bed sharing was reported in only 8% of children. Apnea episodes were observed in 30 infants (42.25 of whole group), in 20 of them only during the neonatal period. More apnea episodes were present in premature infants (48% of premature versus 30% of full term infants). Only 12 children (16.9% of whole group) manifested apnea events during the monitoring period. However, almost all the parents (97%) had a high opinion of the role played by the monitor (mean monitoring time 7.12 months, SD 4.44). In 27 families (38%) a cause of monitoring was a preceeding apnea event. The decision of remaining parents was based exclusively on fear. This fear was justified only in a small number of families (previous SIDS victim, GER). Numerous children were monitored solely on the basis of information of apnea existence in children. Such a difficult to accept monitoring reason probably has its source in inappropriate family health education. PMID- 11013860 TI - [Endoscopic evaluation of upper gastrointestinal tract mucosa in children with food hypersensitivity]. AB - The authors compared clinical symptoms with morphological and endoscopic picture of upper gastrointestinal tract mucosa in 48 children at pre-school and school age with established food allergy. PMID- 11013861 TI - [Production of gastrointestinal mucosa cytokines in Helicobacter pylori infection]. AB - The authors determined the role of cytokines in the pathogenesis of gastric mucous membrane inflammation in patients with H. pylori infection. Cytokines play an important role in the pathogenesis of mucous inflammation in secretory and gastromotoric disorders. PMID- 11013862 TI - [Preoperative diagnosis and surgical treatment tactics for anorectal malformations in light of personal observations]. AB - The article presents contemporary views on classification, diagnostics and surgical treatment of congenital anorectal anomalies. The model of one stage or multi stage repair depending on the kind of defect and the necessity the anal sphincter muscles reconstruction was discussed. It was noticed that the most important independent factor which can have influence on the functional outcome is a normally developed sacral bone. The objective method of evaluation of the sacrum proposed by Schneider Hospital in New York is presented. PMID- 11013863 TI - [The significance of iron in infant development and nutrition. I. Metabolism of iron]. AB - The role of iron in the child organism, as well as the blood's role in regulating the iron homeostasis is described on the basis of relevant literature. From developmental data on Warsaw children and from relevant literature, the child average body iron content and the child average daily iron requirement, which is at age 0-0.5, 0.5-1 and 1-2 years 0.51, 0.92 and 0.76 mg for boys and for girls 0.42, 0.86 and 0.74 mg respectively, are calculated. It is noted that the daily physiological iron loss during the first two years of life can be as much as 50% of the daily iron requirement. Among the causes of infant iron deficiency are the greater blood losses during the peri- and postnatal periods, decreased dietary iron, as well as increased need for iron related to significant body growth. The loss of one ml of blood in children results in the loss of from 0.33 to 0.66 mg of iron. On the basis of the changes in body iron content of infants having various birth weights, it is shown that the quantity of iron needed for proper development is independent of the birth weight. The quantity of iron required by premature infants and children with low birth weight is 25-50% greater, after the infant doubles its birth weight, than that needed by full-term and normal birth weight infants. PMID- 11013864 TI - [The significance of iron in development and nutrition of infants. II. Prevention of iron deficiency]. AB - The symptoms and consequences of iron deficiency in the child are described as found in the literature. The role of iron in immune processes is presented and it is noted that there is no proof for the theory of the development of bacterial infections as a result of giving oral iron supplements. The influence of breast milk and cow milk on the intestinal uptake and loss of iron is discussed. Feeding infants formulae or milk-substitute formulae after cessation of breastfeeding prevents the increased blood loss in the digestive tract, which results from the introduction of cow's milk. The iron content of selected Polish infant formulae, follow-on formulae and milk-substitute formulae available on the market are given and compared with FAO, WHO, ESPGAN and EC recommendations. Results of the authors' own researches on the iron content of selected Polish powdered whole cow's milks are presented. The measurements, made by atomic absorption spectroscopy (AAS), showed that iron content of whole powdered milks ranged from 0.23 to 1.19 mg/dm3. The values do not vary significantly from those in the literature on the iron content of unboiled cow's milk. It is shown that up to the point of the doubling of birth weight, formula-fed infants should be fed modified milk containing at least 2 mg/dm3 of iron. Thereafter infants should receive milk containing at least 7 mg/dm3 of iron. PMID- 11013865 TI - [Cryptorchidism--current opinions]. AB - Cryptorchidism is the most common developmental disorder of endocrine organs in boys. Incidence depends on the age of the child--is the highest in the premature infants (30%), in term infants 2-5%, in adults 0.3%. Aetiology is still unclear and it is thought to be multifactorial including: dysgenesis of the testis, endocrine abnormalities and anatomical block on the way of the descent of the testis. In 90% of cases the pathology is unilateral. Abnormal localisation of the testis leads to the histomorphological changes which may be the causes of complications. The most dangerous are: infertility and neoplasms. The author presents also the diagnostic methods and therapeutic management in cryptorchidism. PMID- 11013866 TI - [Evaluation of total energy balance and food habits of obese children]. AB - Total daily energy expenditure (TDEE) and time spent for activities of different intensities (heart rate monitoring method), as well as dietary and food habits (24-hour dietary recall) in two groups of 12-14 year-old boys, including 35 obese (weight 67.0 kg +/- 9.3, height 162 cm +/- 7.6) and 35 control group boys (weight 46.0 kg +/- 6.5, height 160 cm +/- 7.5) were estimated. Obese boys spent more time at rest and less time on physical activities than nonobese (p < 0.001). That resulted in differences of TDEE expressed per unit of body mass (p < 0.001). Time for sleeping and time in school was not statistically different in the two groups. The evaluation of nutritional habits indicated that the obese boys, when compared with controls, had higher energy intake and significantly higher percentage of fat energy in total energy intake, low intake of fruits and vegetables and irregular frequency of meals during the day. Diets of boys were not balanced with respect to calcium and phosphorus. PMID- 11013867 TI - [Evaluation of iron status in women with pregnancy complicated by tobacco smoking]. AB - Relatively common iron deficiency in pregnant women is assumed to be enhanced by cigarette smoking. In the presented studies we determined cotinine in serum and urine of 75 pregnant women in order to select groups of smoking women and tobacco abstinence. In the smoking group, a mean concentration of cotinine 1039 +/- 560 mg/L in serum and 1025 +/- 540 mg/L in urine were observed. For assessment of iron status we determined in serum: iron, iron-binding capacity (TIBC), transferrin, transferrin saturation, soluble transferrin receptor and ferritin. In serum of smoking woman in comparison to non smoking, the level of ferritin and transferrin was higher but non significantly. Significant increase of TIBC (p < 0.05) and decrease of transferrin saturation (p < 0.05) was observed. Therefore iron deficiency in transport compartment can not be excluded. The concentration of soluble transferrin receptor was the same in both groups studied. However, in late pregnancy (above 27 week of gestation) ferritin concentration less than 20 mg/L of serum was observed in 70% of smoking and only in 39% of non smoking women (p < 0.05). We concluded that cigarette smoking during pregnancy did not have any effect on the entry of iron-bearing transferrin to cells mediated by soluble transferrin receptor, but affected the level of iron-storage ferritin, which leads to iron deficiency in the storage compartment (ID I). PMID- 11013868 TI - [Molecular diagnosis: principles, tasks and purposes]. AB - Molecular diagnosis is a relatively young discipline and also one of the most dynamic among clinical laboratory sciences. Molecular genetic diagnosis is the detection of pathogenic mutations in DNA and RNA samples prepared from at-risk patients. The paper reviews some problems connected with molecular technicqes, organization of a reference laboratory and quality assurance system. PMID- 11013869 TI - [Analysis of mutations in the CFTR gene in patients diagnosed with cystic fibrosis in Poland]. AB - Polish CF patients were screened extensively for mutations in the CFTR gene. Screening data demonstrated a high heterogeneity of CFTR mutations in the Polish population. Total 30 different mutations were characterised in 24 exons or introns of the gene. Among them, six mutations have been reported for the first time and submitted to the CF Genetic Analysis Consortium. In addition, 15 different polymorphisms were found, including three new ones. The screening resulted in 9% increase of the detection rate of CFTR alleles in the tested population. Frequencies of two of the identified mutations (CFTRdele2,3 and 2184insA) are relatively high (2.6% and 1%, respectively) and justify their inclusion into routinely screened mutations in genetic testing of Polish CF population. PMID- 11013870 TI - [Secular growth trends in children and youth of Warsaw in the last twenty years]. AB - The aim of the study was to preliminarly evaluate the secular trend in Warsaw children and youth during the last 20 years. Mean values of body height, weight and chest circumference and weight to height percentile curves of children and youth aged I month to 18 years, examined in Department of Growth and Development of Children and Youth of the Institute of Mother and Child in Warsaw in 1976-80 and 1996-99, were compared. Secular changes in body height and weight and especially in chest circumference were observed in the studied population during the last 20 years. In children and youth growth acceleration occurred, but in infants a deceleration was observed. At all ages a slight tendency toward slimness was visible. The changes in the values of anthropometric growth indices, which occurred during the last 20 years have shown that there is a need for prospective growth monitoring and upgrading of growth standards. PMID- 11013871 TI - [Evaluation of sexual initiation circumstances in youth]. AB - The age of sexual initiation is connected with many factors such as: the age of biological maturity, temperament, personality, moral standards, family, culture and place of residence, religion, influence of partner. The aim of the study was evaluation of the age and circumstances of sexual initiation of youths. The material were voluntary and anonymous questionnaires. These questionnaires included multiple choice questions as well as fill in the blank questions. The questionnaires were given to high-school students. The ages of the 1374 questioned students were between 18 and 20. RESULTS: The age of sexual initiation was between 16 and 18 years of age. The most frequent place was bed at home (50.23 to 67.74%). CONCLUSIONS: The age of sexual initiation does not differ from other European countries. Circumstances of sexual initiation of Poles is similar to other countries. PMID- 11013872 TI - [Idiopathic headache in children]. AB - According to literature data, headaches occur in about 40% of preschool children and up to 70% of the children at school age. Most of the headaches, especially in school children, are idiopathic. Symptomatic headaches accompanying a diversity of diseases occur in a small percentage. The idiopathic headaches are the ones in which the pain is not only the main symptom but also the essence of the disease itself. Other idiopathic headaches include: migraine, tension-type headache and Horton headache. The diagnostic and therapeutic problems in idiopathic headaches are discussed. PMID- 11013873 TI - [Advances in treatment of congenital gastroschisis based on personal examinations]. AB - The aim of the study was the definition of standard approach to a newborn with gastroschisis. The rules of treatment introduced in 1992 enabled the closure of the abdominal wall primarily in all newborns and made it possible to decrease the mortality considerably. This was achieved despite the fact that we usually did not have influence on time and method of delivery or quality of transportation to our department. In the Department of Pediatric Surgery of Institute Mother and Child, in the years 1992-1997, twenty three newborns with gastroschisis were admitted. The standard approach in all patients was the following: insertion of the catheters to the small bowel through the appenddicocaecostomy and to the large bowel through the rectum, intraoperative irrigation of the meconium (mucosolvan), forceful stretching of the abdominal wall, returning of extraabdominal viscera under control of central venous pressure or middle airway pressure (anesthesiologic control). In the postoperative period all patients routinely had controlled ventilation, administration of analgesic drugs, catecholamines, antibiotics and total parenteral nutrition. Intestinal catheters were gradually removed every day for irrigations of all levels. Oral nutrition was started after the return of effective peristalsis. From 23 treated newborns 16 are alive. 7 children died, 4 of them were resuscitated before operation because of severe general condition. Two patients died because of sepsis in the second month of life, one because of SIDS (sudden infant death syndrome) in the third week of life. Sixteen children are followed up. The development of all children is normal. PMID- 11013874 TI - [Hearing assessment in children with Pierre-Robin syndrome]. AB - Physical examination of the ear and hearing assessment was carried out in 30 infants with Pierre-Robin syndrome, aged 3 to 8 months. In 93.3% dysfunction of the Eustachian tube was present. Hearing impairment of perceptive type was found in 13.3% of these children. PMID- 11013875 TI - [Nitric oxide (NO)--Nobel prize in medicine and physiology for 1998]. AB - On October 12, 1998. The Nobel Assembly announced the award of the Nobel Prize in Medicine and Physiology to pharmacologists Robert Furchgott, Louis Ignarro, and Ferid Murad. The Nobel Committee decided to award the prize for their discoveries concerning--nitric oxide as a signalling molecule in the cardiovascular system. Nitric oxide (NO) has a key importance for vascular tonus, acts as a signal molecule in the nervous system and plays an important function in the immunological system. Nitric oxide is a multifunction molecule which controls the blood pressure, modulates gastrointestinal motility. It is produced in abnormal level intensifies septic shock and destruction of nervous tissue. NO is important in different branches of medicine. For instance NO gas has been used to reduced high blood pressure in the lung of infants. Several unknown NO applications in medicine are waiting for discovery. PMID- 11013876 TI - Rice in deep water: "how to take heed against a sea of troubles". AB - Plants are aerobic organisms for which oxygen shortage poses a severe problem. Waterlogging and flooding are the main causes of anaerobiosis and can lead to damage or even death of the plant. Rice is well adapted to semi-aquatic conditions. It is the only cereal that can be grown in flooded areas such as the great river deltas of Asia. In rice, two major strategies have evolved to cope with conditions of flooding. One is to escape submergence and thereby avoid anaerobiosis as much as possible. This is achieved through elongation growth and through extensive aeration of submerged plant parts by way of internal and external air spaces. The second adaptation is a metabolic one which includes the efficient use of carbohydrate resources and maintenance of energy charge when the cells do become anaerobic. The mainly ethanolic fermentation pathway found in anaerobic rice avoids acidification of the cytoplasm and thereby contributes to the maintenance of cell integrity. Genetic analysis indicates that the submergence tolerance trait, which is based on metabolic changes, is encoded by only one or a few as yet unidentified gene(s). Identifying these genes is a major goal in anaerobic stress research. PMID- 11013877 TI - Flavonoid wing pigments increase attractiveness of female common blue (Polyommatus icarus) butterflies to mate-searching males. AB - Common blue butterflies (Polyommatus icarus) sequester flavonoids from their larval host plants and allocate these UV-absorbing pigments to the wings. In field experiments using dummies constructed from female butterflies, mate searching males inspected flavonoid-rich dummies more intensively than those with little or no flavonoids. Flavonoid content as signalled by UV-wing pattern may indicate ontogenetically determined female quality or enhance detectability to males. PMID- 11013878 TI - Walking on insect paths? Early ommatidial development in the compound eye of the ancestral crustacean, Triops cancriformis. PMID- 11013879 TI - Predator odour and its impact on male fertility and reproduction in Phodopus campbelli hamsters. AB - This study investigated the influence of cat urine odour in suppressing development and fertility in Campbell's hamster males. Exposure to this odour from postnatal day 11 until day 45 (sexual maturation) resulted in reduced sex organ weights, reduced testosterone levels and in an increase in abnormalities of the synaptonemal complex in both sex chromosomes and autosomes. Subsequent breeding experiments revealed a significant decrease in litter size. All these data indicate a severe effect of predator odour on the breeding success of potential prey species. It is assumed that these effects are caused by the sulphurous compounds in the urine; however, the underlying mechanisms are not yet known. PMID- 11013880 TI - Free-flight phonotaxis in a parasitoid fly: behavioural thresholds, relative attraction and susceptibility to noise. AB - The phonotactic capacity of tachinid flies to acoustically detect and localize a sound source simulating their cricket host was investigated in a large flight room. Acoustic measurements were performed to estimate the actual stimulus delivered to the flies, revealing highly heterogeneous sound fields. When presented with a simulated cricket song in red or infrared light conditions, the flies readily flew to the sound source and landed on it. Behavioural phonotactic thresholds were established as a function of carrier frequency and were found to coincide well with the frequency of the host's natural song (4.5-5.2 kHz). Experiments revealed that the same range of frequencies is preferentially attractive to the free-flying flies, and that the reliability of signal detection in the presence of noise is best at behaviourally relevant frequencies. PMID- 11013881 TI - A trail pheromone component of the ant Mayriella overbecki Viehmeyer (Formicidae: Myrmicinae). AB - The myrmicine ant Mayriella overbecki lays recruitment trails during foraging and nest emigrations. The trail pheromone originates from the poison gland. From ten identified components of the poison gland secretions only methyl 6 methylsalicylate 1 elicited trail following behavior. PMID- 11013882 TI - Queens and major workers of Acanthomyrmex ferox redistribute nutrients with trophic eggs. AB - In Acanthomyrmex ferox, two distinct egg-types are produced: reproductive eggs that give rise to offspring and trophic eggs that serve to distribute nutrients. Queens lay both reproductive and trophic eggs, while major and minor workers lay only trophic eggs in the presence of the queen. The queen lays on average 17% of the trophic eggs in a colony, while majors and minors produce 42% and 41%, respectively. The large proportion of trophic egg production by the queen and soldiers is quite remarkable, since ant queens are expected to focus entirely on reproduction and majors, which have a defensive function in many species, to be sterile. PMID- 11013883 TI - The brain as a photoreceptor: intracerebral ocelli in the firefly. AB - This paper deals with the structure and function of the intracerebral ocelli in the caudal area of the brain of the Japanese firefly. A pair of epilaterally placed specialized pigmented organs was found at the caudal ends of the brains of the fireflies Luciola cruciata and L. lateralis. On the basis of light and transmission electron micrographs of both male and female individuals these organs seemed photoreceptive in nature. Intracellular and extracellular recordings were obtained from the intracerebral ocelli of the fireflies with microelectrodes. The physiological evidence revealed that the cells found in the brain were, indeed, photoreceptors. PMID- 11013884 TI - Phylogeny of filamentous ascomycetes. AB - Phylogenetic studies of higher ascomycetes are enhanced by the introduction of molecular markers. Most studies employed sequences of the SSU rRNA gene, but recently data from additional genes (RPB2, LSU rRNA) have become available. Several groups defined by their ascoma-type, such as Pyrenomycetes, are supported while others, like the Discomycetes, appear to be paraphyletic. The Pezizales with operculate asci are basal to other eu-ascomycetes, while other Discomycetes appear to be derived eu-ascomycetes. The reevaluation of classical characters using molecular data is discussed using three examples. Ascus types are often regarded as being of major importance in ascomycete systematics, but prototunicate asci were found to be of poor taxonomic value, since ascomycetes with prototunicate asci are polyphyletic. The independence of the Agyriales, assumed from their morphological characters, is supported by sequence data but the relationship to supposed sister groups remains dubious. The phylogeny of ascolocularous fungi and their circumscription requires further study. While a circumscription based on bitunicate asci can be rejected, it remains unclear whether fungi with ascolocularous ascoma development represent a monophyletic entity. PMID- 11013885 TI - Neurosecretion: peptidergic systems in insects. AB - Insect neuropeptides are produced in less than 1% of the cells of the central nervous system. Despite this, they are important messenger molecules which influence nearly all physiological processes, including behaviour. They can act as transmitters, modulators and classical hormones, and often exhibit pleiotropic functions when released into the haemolymph. The large number of neuropeptides that has been identified from some of the model organisms among insects underlines the complexity of the neurosecretory system; studies about the coordinated actions of these substances are in their preliminary stages. Recent advances in insect neuropeptide research will be reviewed here, concentrating on the distribution of multiple peptide forms in the central nervous system and adjacent neurohaemal organs, and the role of neuropeptides in eclosion behaviour. PMID- 11013887 TI - Orientation and navigation during adult transport between nests in the ant Cataglypis iberica. AB - Cataglyphis iberica is a polydomous ant species in which adult transports between nests are frequently observed. When pairs of workers were captured and released at the same location, the transporters (Ts) field directly towards their destination nest and reached it in most of the cases. The transportees (Te), on the other hand, fled in the opposite direction and only a third of them eventually reached their nest of departure. Additional experiments suggest that this result may be explained by the fact that the Ts ants have a memory of the compass direction of the nest they are heading to and that they adjust their course by using a sequence of memorised landmarks. As regards to the Te, the reversal of their direction of transport seems to be based essentially on celestial cues. PMID- 11013886 TI - Biocompatibility correlation of polymeric materials using human osteosarcoma cells. AB - Metal implants are the preferred materials to generate articular prostheses, plates, or bone pegs in orthopedic surgery. Although titanium and titanium alloys show a relatively good biocompatibility, clinical experience revealed that coating of the metallic implant surface may increase the biocompatibility. In a search for optimum bone implant surfaces, we determined polarity and contact angle parameters of a variety of polymers and substances and correlated the findings in a biocompatibility assay using an in vitro bone cell model. We report that an optimum adherence of SAOS-2 cells to such surfaces and a good vitality for polymers are characterized by water-based contact angles of 80 degrees and 20 degrees for advancing and receding probes, respectively. PMID- 11013888 TI - Enzymatic pattern processing. AB - A table-top prototype has been constructed that uses the enzyme malate dehydrogenase to recognize input signal patterns. The device is controlled by the enzyme in response to injection of Mg2+ used as a signaling substance. Output is monitored spectroscopically. If Mg2+ is injected along either of two signal lines (i.e., if the input signal pattern is 10 or 01) the device emits an output of 1. Injection along neither or both lines results in an output of 0. The enzyme in effect is used as a transform that converts the linearly inseparable exclusive- or problem into a linearly separable problem. PMID- 11013889 TI - Pathological alterations typical of human Tay-Sachs disease, in the retina of a deep-sea fish. AB - Micrographs of retinas from the deep-sea fish Cataetyx laticeps revealed visual cells containing membranous whorls in the ellipsoids of the inner segments resulting from stretching and modifications of the mitochondria membranes and their cristae. These pathological structures seem to be homologous to the whorls observed in retinas of human carriers of Tay-Sachs disease. This disease, a genetic disorder, is found in humans and some mammals. Our findings in fish suggest that the gene responsible can be found throughout the vertebrate evolutionary tree, possibly dormant in most taxa. PMID- 11013890 TI - Light-dependent magnetoreception in birds: the effect of intensity of 565-nm green light. AB - In a previous study, Australian silvereyes tested in autumn under monochromatic 565-nm green light at intensities of 2.1 and 7.5 mW m-2 preferred their normal northerly migratory direction, whereas they showed a significantly different tendency towards northwest at 15.0 mW m-2. Repeating these experiments in spring with silvereyes migrating southward, we again observed well-oriented tendencies in the migratory direction at 2.1 and 7.5 mW m-2. At 15.0 mW m-2, however, the birds once more preferred northwesterly directions, i.e. their response under this condition proved to be independent of the migratory direction. This contradicts the interpretation that monochromatic green light of this high intensity leads to a rotation of compass information; instead, it appears to produce sensory input that causes birds to give up their migratory direction in favor of a fixed direction of as yet unknown origin. PMID- 11013891 TI - Radio-telemetric evidence of migration in the gregarious but not the solitary morph of the Mormon cricket (Anabrus simplex: Orthoptera: Tettigoniidae). AB - The Mormon cricket, Anabrus simplex, is one of just a few species of katydids (or bushcrickets, Orthoptera: Tettigoniidae) that, like migratory locusts, appear to have solitary and migratory morphs. Using radio telemetry we studied movements of individuals of two morphs of this flightless species. Individuals within each migratory band had similar rates of movements along similar directional headings whereas solitary individuals moved little and showed little evidence of directionality in movement. Our results also add to other recent radio-telemetry studies showing that flightless insects of 1-2 g in mass can be tracked successfully using these methods. PMID- 11013892 TI - Seed drops and caches by the harvester ant Messor barbarus: do they contribute to seed dispersal in Mediterranean grasslands? AB - To determine whether the harvester ant Messor barbarus acts as a seed disperser in Mediterranean grasslands, the accuracy level of seed processing was assessed in the field by quantifying seed drops by loaded foragers. In the vicinity of exploited seed patches 3 times as many diaspores were found as in controls due to seed losses by foragers. Over trails, up to 30% of harvested seeds were dropped, singly, by workers but all were recovered by nestmates within 24 h. Seeds were also dropped within temporary caches with very few viable diaspores being left per cache when ants no longer used the trail. Globally, ant-dispersed diaspores accounted for only 0.1% of seeds harvested by M. barbarus. We discuss the possible significance for grassland vegetation of harvester-ant-mediated seed dispersal. PMID- 11013893 TI - 2,3-dimethyl-5-(2-methylpropyl)pyrazine, a trail pheromone component of Eutetramorium mocquerysi Emery (1899) (Hymenoptera: Formicidae). AB - The ant Eutetramorium mocquerysi (Myrmicinae) is endemic to the island of Madagascar. During foraging and nest emigration the ants lay recruitment trails with secretions from the poison gland. We identified three pyrazine compounds in the poison gland secretion: 2,3-dimethyl-5-(2-methylpropyl)pyrazine 1, 2,3 dimethyl-5-(3-methylbutyl)pyrazine 3, 2,3-dimethyl-5-(2-methylbutyl)pyrazine 4. Only the first component elicited trail-following behavior in the ants. We were unable to investigate whether the other pyrazine components have a synergistic function. PMID- 11013894 TI - [The p300 cognitive event-related potential. I. Theoretical and psychobiologic perspectives]. AB - The P300 is a positive wave which arises when an attended stimulus is detected. Its parameters depend on a number of variables, in particular the subject's mental state, the task that has to be accomplished, the significance of the stimulus, and the degree of attention. It can be recorded with accuracy, and the different stages of information processing can therefore be analyzed. The P300 wave shows the modifications in neuronal activity which take place during the cognitive process: P300 latency provides an indirect indication of the duration of the processes involved in stimulus discrimination while its amplitude, which is influenced by a number of variables, provides an index of the intensity of the energetic activation or arousal involved. The P300 wave consists of several components which reflect distinct information-processing events (P3a, P3b, P3e, P SR, P-CR). According to the theoretical models, it is hypothesized that P300 could either represent the adaptation of the working memory to further environmental input ('context updating'), or indicate a closing process ('context closure') in information processing. As regards the physiological aspect of P300 and its association with cortical networks, various studies have suggested that several cortical generators of P300 may co-exist: the medial temporal lobe, the temporo-parietal junction, and the medial and lateral frontal lobe. Psychopharmacological studies have shown that different neurotransmitter systems are involved in the generation and modulation of P300, namely the cholinergic, noradrenergic, dopaminergic, serotoninergic and gabaergic systems. It appears that the noradrenergic agonists increase the amplitude of P300, dopaminergic agonists may have a biphasic effect (increase/reduction), while cholinergic antagonists and gabaergic agonists reduce P300 amplitude and prolong its latency. PMID- 11013895 TI - [The p300 cognitive event-related potential. II. Individual variability and clinical application in psychopathology]. AB - The P300 wave is one of the cognitive components of the event-related potential (ERP) that is used to investigate the cognitive processes, and which can be used to study patient populations with a variety of psychiatric disorders. Its clinical utility has been increased by the identification of factors that contribute to the variability in its amplitude and latency. However, its value as a diagnostic index has not been entirely established. It can provide a useful recording of patients' information processing, and indicate the severity of the clinical state and its possible evolution. It can also assist in determining what therapeutic approach to adopt. In the present review, the findings in the literature concerning interindividual variation in the P300 wave are first described; several variables significantly influence the amplitude and latency of this wave, such as age, gender, intelligence and personality. Following this, the relevance of the data in the literature on the clinical applications of P300 in psychopathology is examined, including the studies undertaken to obtain an objective diagnostic index for mental disorders and also those carried out to assess the problems concerning the interpretation of information connected with the mental pathologies examined. P300-associated findings on dementia, schizophrenia, depression, alcoholism, drug addiction, anxiety disorders (panic disorder, obsessive-compulsive disorder, and post-traumatic stress syndrome) and on personality disorders (schizoid, antisocial or borderline personality disorder) have been examined in detail. PMID- 11013896 TI - The effect of motivational instructions on P300 amplitude. AB - The aim of this investigation was to determine the effect on P300 amplitude of instructions aimed at increasing the subject's degree of task involvement. To this end, two different studies were carried out. In Study 1, 20 university students were tested with an auditory event-related potential (ERP) oddball paradigm (target: 1,100 Hz; standard: 1,000 Hz) in two consecutive runs, each with a different set of instructions; after the first run, subjects were verbally motivated to increase their level of performance in the second run. In Study 2 (performed 1 year later), ERPs were similarly obtained from the same subjects during two oddball runs, but this time both tests were preceded by neutral instructions. The amplitude and latency of N1 and P2 elicited by non-targets and of N2 and P3 in target waveforms were evaluated. The findings showed that following motivating instructions, P3 amplitude increased while P3 latency showed a non-significant decrease. The amplitude of P2 to non-target stimuli--which could be interpreted as P250--was also affected by the instructions provided. The overall results suggest that the presentation of motivating instructions is followed by a higher amount of attentional resources allocated to all stimuli, and a more efficient evaluation and discrimination of relevant targets. The implication of these findings for the clinical use of P300 has been discussed. PMID- 11013897 TI - Predictive value of EEG monitoring during drug withdrawal in children with cryptogenic partial epilepsy. AB - In order to evaluate the predictive value of the interictal EEG after discontinuation of anticonvulsant therapy in children with cryptogenic partial epilepsy, a prospective study was carried out on 84 children and adolescents who had been seizure-free for at least 2 years. Twenty-four children (28.6%) presented one or more relapses. EEG monitoring during drug withdrawal showed abnormalities in 67% of the patients (16/24) who relapsed with seizures, and in 10% of the patients (6/60) who did not relapse. Our study suggests that in children who have suffered from partial epilepsy, the detection of paroxysmal abnormalities during drug withdrawal, in a previously normal EEG is a risk factor for recurrence of seizures. It is therefore advocated that periodic EEG monitoring be carried out during the drug withdrawal period, because the presence of EEG abnormalities is associated with a high probability of seizures occurring during or after drug discontinuation. PMID- 11013898 TI - [Evoked mechanical and electrical anal sphincter responses after cortical and lumbar magnetic stimulation]. AB - In 13 healthy volunteers we recorded electrical and mechanical anal sphincter (AS) evoked responses. Electrical responses were obtained with surface electrodes, and mechanical responses by anal manometry; all responses were compared with those of anterior tibialis. Mechanical evoked responses' latency and amplitude were calculated. The mean total conduction time of the electrical responses was 26.07 ms at rest and 23.31 ms with mild contraction (p < 0.003). After lumbar magnetic stimulations, because of stimulation artefact, electrical responses couldn't be correctly measured. The mean central conduction time calculated with mechanical responses was 21.44 ms at rest and 18.81 ms with mild contraction (p < 0.003). The mean central conduction time to the anterior tibialis was shorter (14.38 ms; p < 0.003). Evoked mechanical responses', amplitudes were respectively 128.2 cm H2O (cortical stimulations at rest), 138.8 cm H2O (cortical stimulations with mild contraction) and 133.1 cm H2O (lumbar stimulations at rest) (NS). Evoked mechanical responses amplitudes were correlated with the AS maximal voluntary contraction amplitude (p < 10(-4)). Mechanical motor evoked responses after cortical and lumbar magnetic stimulations can be recorded and quantified. In anorectal disorders, this technique could be of major interest for the diagnosis of neurologic lesions versus behavioural problems. PMID- 11013899 TI - Expanding the reach of clinical trials. PMID- 11013900 TI - Reader looking for guidelines/policies on continuous infusion vesicants for assessments. PMID- 11013901 TI - Additional issues raised on blood returns and ports. PMID- 11013902 TI - Thyroid cancer: patients' experiences of receiving iodine-131 therapy. AB - PURPOSE/OBJECTIVES: To enhance understanding of the experiences and educational needs of patients receiving iodine-131 (131I) therapy for the treatment of thyroid cancer. DESIGN: Qualitative design using interpretive description. SETTING: A 24-bed oncology/acute-care medical unit in a large tertiary hospital in western Canada. SAMPLE: 5 men and 22 women (18-80 years of age) who have received 131I therapy in the past two years. METHODS: Unstructured focus group and telephone interviews and field notes. Interviews were tape-recorded, transcribed verbatim, and subjected to thematic analysis. FINDINGS: Researchers elicited four major themes from the analysis: (a) recognizing the totality of the cancer experience, (b) being isolated, (c) recognizing the totality of the treatment experience, and (d) understanding barriers to treatment. CONCLUSIONS: Healthcare providers require a better understanding of thyroid disease and the treatment and side effects of 131I therapy. Furthermore, educational programs are required to adequately prepare nurses and patients for future care. IMPLICATIONS FOR NURSING PRACTICE: A need exists to improve the care and education provided to patients receiving 131I therapy. In particular, recognition of the totality of the cancer experience and the need for both staff and patient education were illustrated. If nurses are to provide comprehensive cancer care, both psychosocial and physical needs must be addressed and fulfilling these needs requires a collaborative approach among patients, nurses, and other healthcare professionals. PMID- 11013903 TI - Current treatment approaches for neoplastic meningitis: nursing management of patients receiving intrathecal DepoCyt. AB - PURPOSE/OBJECTIVES: To review neoplastic meningitis and the nursing implications for the intrathecal administration of chemotherapy. DATA SOURCES: Published research and educational manuscripts, books, conference proceedings, and personal experience. DATA SYNTHESIS: Standard treatment for neoplastic meningitis includes radiotherapy to the neuraxis to palliate symptomatic disease, intrathecal chemotherapy to eradicate cancer cells in the cerebrospinal axis, and systemic chemotherapy. Intrathecal liposomal cytarabine (DepoCyt), a novel lipid encapsulated chemotherapeutic agent, prolongs tumor exposure to cytotoxic levels of cytarabine, improves patient response rates, and prolongs time to clinical progression. CONCLUSIONS: DepoCyt prolongs the half-life of cytarabine in the central nervous system, resulting in improved patient response to therapy and delayed disease progression. IMPLICATIONS FOR NURSING PRACTICE: The clinical success of DepoCyt treatment depends on effective implementation of the treatment regimen, attentiveness to patient and family education, and adverse-event management. PMID- 11013904 TI - An international feasibility study of parental decision making in pediatric oncology. AB - PURPOSE/OBJECTIVES: To describe parental decision making about treatment options for children with cancer and determine the feasibility of a similar but larger international study. DESIGN: Exploratory. SETTINGS: A pediatric catastrophic illness research hospital in the United States and children's hospitals in Australia and Hong Kong. SAMPLE: A convenience sample of 43 parents (5 fathers and 38 mothers ages 23-59 years). METHODS: Six open-ended interview questions posed to parents during private individual interviews. Content analysis techniques were used. MAIN RESEARCH VARIABLES: Parental perceptions of (a) factors considered in the decision-making process, (b) behaviors of healthcare professionals that affected the process, and (c) satisfaction with the process. Feasibility of a larger study was estimated by considering ease of access to parents, number of refusals to participate, understanding of the interview questions, and level of interest at each setting. FINDINGS: Access to parents was possible at all sites. Refusal to participate was reported only at the U.S. site. Certain factors (e.g., getting information from the healthcare team, trusting staff) were important to all parents considering end-of-life decisions. Site specific factors included considering alternative therapies (at the Australian site) and strengthening faith (at the U.S. site). CONCLUSIONS: A larger international study of parental decision making is feasible. Sufficient similarities in parental decision making exist across these sites to justify future efforts to identify universal decision-making factors that, in conjunction with site-specific differences, could be helpful in developing guidelines for healthcare professionals who assist parents in making treatment-related decisions for a sick child. PMID- 11013906 TI - Men's perspectives on the impact of prostate cancer: implications for oncology nurses. AB - PURPOSE/OBJECTIVES: To describe the perspectives of men with recurrent prostate cancer regarding their experiences with the disease, its impact, and the help they received and to compare these individuals to men without recurrent prostate cancer. DESIGN: Cross-sectional survey. SETTING: Physicians' offices and prostate cancer self-help groups in Canada. SAMPLE: A convenience sample of 120 men with recurrent prostate cancer and 845 men without recurrent disease. METHODS: A survey instrument was developed following in-depth interviews with men living with prostate cancer. The men received survey packages from their physicians or through a self-help group, completed the instrument at home, and returned it in a prestamped addressed envelope. MAIN RESEARCH VARIABLES: Impact on lifestyle, satisfaction with communication, importance of information, satisfaction with information received, problems experienced, and assistance received for problems. FINDINGS: Significant differences related to a number of factors were found between the men with recurrent prostate cancer and those without recurrent disease. A large number of those with recurrent disease experienced problems with side effects, anger, and pain and received help for their pain; experienced a negative impact on leisure time and on mental health; experienced difficulty talking with healthcare professionals; and felt the need to talk with someone about their cancer. Many of the men with recurrent disease were dissatisfied with the information they received about their medical condition and possible side effects. CONCLUSIONS: Men with prostate cancer experience both physical and psychosocial difficulties. Many perceive that they are not receiving adequate help for these difficulties. Future research is needed to increase understanding of how men are managing the impact of prostate cancer and what types of interventions would be most useful. IMPLICATIONS FOR NURSING PRACTICE: Findings emphasize the need for nurses to be certain that they are conducting broad-based assessments of patients with prostate cancer throughout the illness experience. Nurses need to inform patients about services available to assist them, help patients understand the emotional responses to illness, and provide opportunities for patients to talk about the illness and its impact. PMID- 11013905 TI - Implementing the Agency for Health Care Policy and Research's Smoking Cessation Guideline in a lung cancer surgery clinic. AB - PURPOSE/OBJECTIVES: To determine the effectiveness of a nurse-managed smoking cessation intervention based on the Agency for Health Care Policy and Research's (AHCPR's) Smoking Cessation Guideline in a lung cancer surgery clinic. DESIGN: Quasi-experimental. SETTING: Urban, Midwest, academic, and tertiary care. SAMPLE: 25 adult male and female smokers with a confirmed diagnosis of lung cancer that had been surgically managed. Subjects were assigned to an intervention group (n = 14) or a usual-care group (n = 11). METHODS: Participants in the intervention group received a nurse-delivered, AHCPR-based smoking cessation intervention that included face-to-face and phone follow-up contact beginning with the first preoperative clinic consultation. Usual-care participants received routine care provided at the institution. MAIN RESEARCH VARIABLES: Self-reported smoking status with expired air carbon monoxide confirmation six months postsurgery. FINDINGS: Seventy-one percent of the intervention group was biochemically confirmed to be abstinent by expired air carbon monoxide, as compared to 55% in the usual-care group. CONCLUSIONS: Smokers diagnosed with lung cancer desired to quit smoking and may benefit from an intensive smoking cessation intervention at time of diagnosis. IMPLICATIONS FOR NURSING PRACTICE: Further research should include continuing evaluation of an intensive smoking cessation intervention with this population, and all clinicians should be trained to implement AHCPR's Smoking Cessation Guideline in practice. PMID- 11013907 TI - Exploring the information flow: partners of women with breast cancer, patients, and healthcare professionals. AB - PURPOSE/OBJECTIVES: To explore the information flow between partners of women with breast cancer and patients and between partners and healthcare professionals (HCPs). DESIGN: Descriptive. SETTING: Community-based. SAMPLE: 109 partners of women with breast cancer. METHODS: Subjects completed a 30-item, self administered questionnaire that was designed to identify the informational flow between them and their information providers. Subjects also completed the Miller Behavioral Style Scale to determine the informational coping styles of individuals under threat. MAIN RESEARCH VARIABLE: Informational flow between partners and patients and partners and HCPs. FINDINGS: The majority of partners received information from patients and were satisfied with the amount of time they spent discussing breast cancer topics. Few partners thought that patients avoided giving them information. A significant association existed between the informational coping styles of partners and the communication flow between partners and patients. The majority of partners sought information from other sources. In addition, the majority of partners accompanied patients to their medical consultations and received information from HCPs. The communication flow among partners, patients, and HCPs largely depended on the amount of communication desired by the woman with breast cancer. CONCLUSIONS: This study provides new evidence on the information flow and communication patterns between partners and patients and between partners and HCPs. IMPLICATIONS FOR NURSING PRACTICE: Partners may have informational needs that are not met by patients. Therefore, nurses need to provide them with information to help satisfy their needs. In addition, nurses can help satisfy the informational needs of partners by encouraging good two-way communication and informational exchange between partners and patients. PMID- 11013908 TI - Assessment of patient satisfaction with pain management in small community inpatient and outpatient settings. AB - PURPOSE/OBJECTIVES: To describe patient outcomes (e.g., pain intensity and relief, satisfaction, expectations) and analgesic practices of healthcare providers for inpatients and outpatients in community hospital settings. DESIGN: Descriptive, correlational, and random sampling. SETTING: Three community-based institutions in southeast Louisiana. SAMPLE: 114 inpatients and outpatients with cancer-related or acute postoperative pain. Inpatients (n = 68) mostly were women and younger than 60 years of age. Outpatients (n = 46) mostly were men and older than 60 years of age. Both groups were predominantly well-educated and Caucasian. METHODS: Subjects completed a modified version of the American Pain Society's Patient Satisfaction Survey. Researchers completed a chart audit tool reviewing analgesic prescriptive and administrative practices. FINDINGS: Weak to moderately strong correlations existed for the relationships between the satisfaction variables and the pain intensity, pain relief, and expectation variables for all subjects. Satisfaction with current pain intensity was correlated most strongly with pain intensity and relief scores. Higher pain intensity and relief were related to lower satisfaction with current pain intensity. CONCLUSIONS: Regardless of setting or pain type, subjects experienced significant amounts of pain during a 24-hour period. Patient expectations for experiencing high levels of pain were realized, but expectations for significant pain relief were not. IMPLICATIONS FOR NURSING PRACTICE: Institutional pain management programs that approach pain from a multidimensional perspective need to be developed. Continued education for healthcare professionals and patients is a vital part of this process. PMID- 11013909 TI - [The right way to the future of knee endoprostheses]. PMID- 11013910 TI - [Orthopedics, orthopedic surgery, trauma surgery. Coming full circle?!]. PMID- 11013911 TI - [Knee endoprostheses: problems and technological developments from the manufacturer's point of view]. AB - Modern-day knee arthroplasty is based principally on the experience gained in the course of the past three decades. In the 1980s and 1990s, for example, the full constraint models of the 1970s were replaced by surface replacement prostheses with a combined rolling/sliding motion approximating the natural motion of the human knee joint for a wide range of applications. Not all of the relevant questions in the areas of material science, biomechanics, tribology, implant fixation and surgical technique have been fully explored by any means. Nevertheless, the medium-term and long-term results presently attained by knee arthroplasty are comparable to those achieved in the field of hip-joint arthroplasty. Although the success achieved by today's knee-joint prostheses is impressive, a critical analysis of therapeutic failures and complications is necessary to discover areas where further research is necessary and to formulate the challenges presenting themselves in R&D and in the manufacture of artificial knee joints. Currently, research efforts are focused on optimizing materials (especially in light of the presently unsatisfactory wear behavior of artificial knee joints), biomechanically-assisted design optimization to achieve a better harmonization between mobility and stability in the knee joint to be treated, alternative anchoring concepts, including consistently cement-free anchoring, and solutions for simplifying implantation techniques and making them more precise. The latter area, in particular, appears to be of great importance, since the quality of artificial knee joints and the long-term therapeutic outcome of knee joint arthroplasty depend to a great extent on the correct alignment of the leg axes, an optimal implant position, and the correct adjustment of soft tissue tension. Technical innovations in this area of implant technology exert a multifactorial impact on different aspects of arthroplasty--e.g., uniform force transfer to minimize wear, selective rotation of the femoral components to optimize patella tracking, and avoidance of load concentrations to improve fixation strength--and thus form the basis for a surgical approach that takes greater account of "the big picture". Today's sophisticated navigation systems are a case in point. The non-imaging systems, in particular, have proved to be exceptionally useful; their advantages lie in their simple handling which makes use of common surgical techniques, the avoidance of additional imaging data recording (e.g. CT images), and an inconsequential increase in operation times. PMID- 11013912 TI - [Knee endoprosthesis: aspects of surgical techniques]. AB - The implantation of a condylar knee has remained a challenge for surgeons and the equipment of the hospital. In contrast to total hip arthroplasty, not only is the correct insertion of the implant crucial, but also the treatment of the surrounding soft tissues. Mediolateral soft tissue balance and balance between flexion and extension gaps, as well as centering the patella, have to be done carefully. The soft tissue envelope of the knee joint is thin and prone to necrosis after multiple incisions. Damage to the extension mechanism can make a total joint useless. Tearing the patellar ligament from the tibial tuberosity must therefore be avoided by all means during operation. Precise cutting of the distal femur is only possible if excellent equipment is used by a skillful surgeon. The same is true for orientating saw cuts related to the long leg axes. These tasks can probably be taken over by navigation systems and robots in the future. Because the definite choice of implants has to be made intraoperatively, a complete modular system has to be present in the hospital. Only hospitals that can properly equip their surgeons to manage upcoming soft tissue problems and bony deficiencies should offer treatment to patients requiring condylar knee arthroplasty. PMID- 11013913 TI - [Knee endoprosthetic surgery: tasks and problems of the surgical instrument teams]. AB - About 220 knee endoprostheses are implanted at our clinic annually. This is a routine procedure for the instrument staff. Practice has shown, however, that deficient operating room management as regards the instruments, implants, operating room staff, and surgical planning causes problems. As a result of this, a prophylactic search for sources of error began (risk management). If the sources of error are found, described, and remedied, knee endoprosthetic surgery will be successful for both the patient and the surgical team. Cooperation and communication among manufacturer, surgeon, and surgical nursing staff should serve to minimize forensic risks. The most important task of the instrument staff is to produce adequate structure and procedural quality so that a positive result can be achieved. PMID- 11013914 TI - [Knee endoprosthesis: anesthesiological considerations]. AB - The anesthetic care of patients undergoing total knee arthroplasty (TKA) is described based on experience with reports of the recent literature. The problems and complications of cooperation between different personnel and speciality groups treating the patient are of a more organizational nature during the preoperative phase, and are aimed primarily at determining the appropriate scheduling of the operation. At the forefront intraoperatively is the appropriate monitoring of the patient, and the adjustment of anesthetic regimes to the individual needs and requirements of the patient. The obligation for intensive monitoring as well as an appropriate pain therapy are indispensable in the postoperative phase. PMID- 11013915 TI - [Knee endoprosthesis: biomechanical requirements and consequences]. AB - The experiments described herein permitted the dynamic in vitro measurement of the mechanical efficiency of the quadriceps muscle, both before and after total knee arthroplasty (TKA). The measurement of tibiofemoral pressure on fixed and mobile polyethylene inlays, as well as the movement of mobile inlays, is described. The Interax TKA system was implanted in 8 fresh frozen knee cadavers which were then mounted onto a knee-simulating machine. An isokinetic extension motion was simulated by extending the knee specimens under a constant extension torque of 31 Nm. The quadriceps muscle force required for this motion was not found to increase after implantation of the prostheses. Pressure on the mobile inlay was 60% lower than that measured on the fixed inlay. A 4-mm posterior migration of the mobile inlay on the tibial base plate was observed, with no motion of the inlay femoral component contact surface, which migrated posteriorly in the fixed bearing prosthesis. Thus, instead of moving posteriorly, the mobile inlay femoral component contact area remained at the center of the inlay during the entire extension motion. PMID- 11013916 TI - [Knee endoprosthesis: selection and requirements of materials]. AB - The possibilities of using different material combinations for the replacement of knee joint endoprostheses are very limited. Therefore, cobalt-chromium-molybdenum alloys are currently being used for femoral components and ultra-high molecular weight polyethylene for tribologically stressed tibial components. Titanium alloys can be considered for the tibial component only. For tribological reasons, it is not possible to use a femoral component made of titanium unless it has a corresponding coating. As far as the design is concerned, problems arise from the fact that, on the one hand, there is a demand for the smallest possible size or resection height. On the other hand, however, the forces and strains are rather high and therefore a certain material thickness is necessary in order to avoid fatigue fracture. Regarding polyethylene, the same known principle must be taken into consideration here--not to avoid using the so-called floating design, i.e., the polyethylene components should always have a supporting limitation, or cold flow might occur. The tribological behaviour of polyethylene is restricted in cases of constrained tibia plateaus in so far as the linear or punctual contact of the initial run-in phase leads to correspondingly high surface pressure consequently overstressing the polyethylene. In order to improve this, mobile meniscal bearings are used and the surface pressures achieved here can be endured by the polyethylene even over a long-term. The extent to which the new so-called cross-linked polyethylene can be used in knee joint endoprosthetics is currently being tested, and the simulator results in this respect have been promising so far. PMID- 11013917 TI - [Knee endoprosthesis: clinical aspects]. AB - Despite improvements in component design, instruments, and operative technique, there remains a significant complication and failure rate in total knee arthroplasty (TKA). Revision TKA accounts for every tenth operation in TKA. From June 1991 to June 1994, 209 TKAs (182 patients) were performed. Of these, 73% were uncemented, 11.2% cemented, and 15.8% hybrid. The patella was resurfaced in 96% with a cemented polyethylene or an uncemented metal-backed patella component. Follow-up showed significant improvement in knee and function scores 1 and 5-7 years postoperatively. Of the patients, 77% showed no general and 80% no local postoperative complications. Overall, 42 revisions (41 patients) were performed during the 7-year follow-up. Revision surgery was necessary because of aseptic loosening (8.1%), polyethylene wear (4.8%), complications of the extensor mechanism (2.4%), traumatic periprosthetic fractures or knee luxation (1.4%), and septic loosening (3.3%). Further improvements of polyethylene will reduce polyethylene wear. Uncemented fixation of this prosthesis can only be recommended with hydroxyapatite coating. PMID- 11013918 TI - [Knee endoprosthesis: sports orthopedics possibilities and limitations]. AB - Many patients would like to resume some sport activities after total knee replacement; however, most recommendations are based on subjective opinion rather than scientific evidence. The following paper presents a literature review of sports after total knee replacement and includes recommendations which are based on biomechanical laws. Most total knee designs show increased conformity near full extension. Beyond a certain knee flexion angle, the conformity ratio decreases due to a reduced femoral radius. Therefore, these designs accept higher loads near full extension than in flexion. In order to recommend suitable physical activities after total knee replacement, both the load and the knee flexion angle of the peak load must be considered. It has been shown that power walking and cycling produce the lowest polyethylene inlay stress of a total knee replacement and seem to be the least demanding endurance activities. Jogging and downhill walking show high inlay stress levels and should be avoided. Hence, for mountain hiking, patients are advised to avoid descents or at least use skipoles and walk slowly downhill to reduce the load on the knee joint. It must also be mentioned that any activity represents additional wear, which may affect the long term results of total knee replacements. Further clinical studies are necessary to validate the biomechanical investigations. PMID- 11013919 TI - [The 48th Annual Meeting of the Society of South Germany Orthopedists, Baden Baden, 28 April-1 May 2000]. PMID- 11013920 TI - Experiences in the realisation of a research project on anthroposophical medicine in patients with advanced cancer. AB - QUESTIONS UNDER STUDY: To date most of the published studies on the effectiveness of complementary therapies in cancer patients have yielded controversial results because of questionable methodology. Research strategies and methodologies acceptable to both conventional and unconventional medicine are difficult to find due to different belief systems. In this publication we describe the development and implementation of a project conducted as part of National Research Programme 34 (NFP 34). Detailed analysis of our experiences might provide some information on how to deal with practical difficulties in the planning and conduct of further research projects in this field. The project involved the anthroposophical Lukas Clinic in Arlesheim and the Institute of Medical Oncology of the University Hospital, Berne. This interdisciplinary research project was devised to study the relative merits of these two schools of medicine in the care of advanced cancer patients. The project was made up of three components: (1) a registration study aimed at comparing the case mix at the two institutions; (2) a three armed randomised study on the effectiveness of supportive therapy, comparing anthroposophy to psychosocial group therapy, and (3) a longitudinal study to monitor the evaluation of quality of life of patients at the anthroposophical clinic. METHODS: After a brief review of the study protocol, which presents the theoretical framework of the project, problems of its implementation are described. Aspects of accrual, acceptance of randomisation and data availability are presented using simple descriptive statistics and logistic regression. RESULTS: The registration study was duly completed with a total of 567 patients. For several reasons (not meeting inclusion requirements, high refusal rate) the accrual into the randomised study was slower than expected and required modification of the original design specifications with regard to inclusion criteria and data collection schedule. Additionally, a high dropout rate contributed to premature closure of this part of the project. The longitudinal study also suffered from low data availability at follow up. CONCLUSIONS: The study protocol constituted a major effort at compromise without loss of scientific rigour, and this effort demonstrates that it is possible to allow for different views on patients, on clinical interventions and on research strategies when establishing collaboration between different schools of medicine. Despite a theoretically sound framework, the randomised part of the project proved difficult in its practical execution. Some unexpected logistical constraints and some unmet expectations influenced the feasibility of this part of the project. Therefore, careful planning of research projects in this field of medicine should always include an extended analysis of various practical aspects of study implementation. PMID- 11013921 TI - Clinical features and analysis of the duration of colonisation during an outbreak of Salmonella braenderup gastroenteritis. AB - During an outbreak of acute Salmonella braenderup gastroenteritis we performed a standardised interview encompassing questions on clinical symptoms in 156 (127 adults and 29 children) of 215 identified patients. Sequential stool cultures were obtained for up to five months in these 156 cases. We restricted the analysis to the 122 patients with at least 3 or more available cultures. They were treated with a fluoroquinolone, trimethoprim-sulfamethoxazole, or not treated with antibiotics, according to the decision of the practitioners. For this reason, a randomised double blind study was not possible. Minimum inhibitory concentrations (MIC) of the prescribed drugs were measured for representative isolates before and after treatment. The most frequent symptoms were diarrhoea (98%) and abdominal pain (96%). Vomiting occurred in 43% of cases. Children were more severely ill. Seven weeks after acute gastroenteritis, stool cultures were still positive for salmonella in 71% of the 22 children and 30% of the 100 adults examined (p < 0.002). This rate decreased progressively in both groups to 5 and 3% respectively at 20 weeks (n.s.). Among adults, no significant difference in enteric carriage over time could be demonstrated between untreated patients and those treated with either a fluoroquinolone or trimethoprim-sulfamethoxazole. MIC for salmonella isolates remained unchanged after treatment. In a cohort of patients infected with a single strain of salmonella, fluoroquinolone therapy of acute gastroenteritis failed to influence the duration of enteric carriage, despite continuing susceptibility of the strain. In children, the rate of clearance of Salmonella braenderup from stool was statistically lower until the tenth week after the acute disease, but there was no further difference after 5 months. PMID- 11013922 TI - [Medical treatment of heart failure: an analysis of actual treatment practices in outpatients in Switzerland. The Swiss "IMPROVEMENT of HF" Group]. AB - BACKGROUND: From several studies in Europe and the USA there is evidence that drug treatment of patients with congestive heart failure (CHF) could be improved. There are only sparse data on the treatment of this population in Switzerland. METHODS: In the context of a European Study (IMPROVEMENT of HF Study), in 1999, the treatment of 474 patients with symptomatic CHF was recorded by chart review with primary care physicians throughout Switzerland. The effect of potential predictors of drug treatment was tested using multivariate logistic regression. RESULTS: Mean age of the study population was 75 +/- 12 years. Overall, angiotensin-converting enzyme inhibitors (ACE-I) and angiotensin receptor blockers (ARB) were prescribed to 65% of the study population. Beta-blockers, loop diuretics/thiazides, spironolactone and digitalis were prescribed to 25%, 73%, 13% and 31% respectively. Compared with CHF patients < 65 years of age, the odds ratio of ACE-I/ARB prescription in patients aged 65-74, 75-84, and > or = 85 years was 0.80, 0.58 and 0.40 respectively (p < 0.001). The respective odds ratios for beta blocker treatment were 0.37, 0.21 and 0.06 (p < 0.001). In addition, NYHA classification, comorbid conditions such as renal failure and contraindications strongly predicted drug prescription. Gender and geographical area were not associated with drug selection. CONCLUSIONS: Overall drug prescription among CHF patients in Swiss primary care appears to be satisfactory. However, prescription of ACE-I/ARB and beta-blockers falls steeply with increasing age, independent of measured comorbid conditions and contraindications. Thus, improvement of treatment should focus on a more consistent use of these drugs in the segment of elderly CHF patients. PMID- 11013923 TI - [Start of an adult living donor liver transplantation program in Switzerland]. AB - The shortage of cadaver organs has prompted transplant centres to seek new sources of grafts. While living-donor left lobe transplantation (segments II and III) is an established procedure for children, living donor right liver transplantation (segments V, VI, VII, VIII), which can provide adequate liver mass for an average-sized adult patient, is technically more demanding and potentially associated with higher risks for the donor. In view of the permanent shortage of organs in Switzerland, we started an adult living donor liver transplantation programme in 1999 with the approval of the Clinical Ethics Committee of Geneva University Hospitals. Donor evaluation was performed only after the recipient had been officially registered for transplantation in the national waiting list. Preoperative evaluation consisted of a preliminary information phase with blood tests and Doppler ultrasonography, a second phase with radiological non invasive investigations (CT scan with volume measurements, magnetic resonance cholangiography) and a third phase including liver biopsy and angiography. A formal psychiatric evaluation was performed in all cases and detailed consent was required. Eight potential donors were investigated, 5 were not retained because of too small right liver or steatosis, and 3 were accepted (wife, son, sister). Living-donor hepatectomy was performed without interrupting the vascular blood flow. The liver graft was perfused ex-situ with University of Wisconsin solution. The grafts were anastomosed to the preserved vena cava of the recipient and the portal and arterial anastomoses were performed without interposition grafts, with short cold ischaemic times in the 3 cases. The graft to-recipient weight ratio ranged from 1.04 to 1.12%. The grafts worked immediately; the post-operative course in the 3 recipients was unremarkable and no rejection episode occurred. Significant complications were observed in one donor (percutaneously drained bilioma and spontaneously resolved popliteal sensory palsy). Living-donor right liver transplantation is a potentially valuable solution to the increasing shortage of donor organs. The procedure can be performed safely provided stringent criteria for donor selection, for donor recipient coupling (> 1% graft to body weight ratio) and for centre selection (experience in liver surgery, reduced and split liver transplantation) are applied. PMID- 11013924 TI - [Expandable esophageal stent for the treatment of esophageal fistula]. PMID- 11013925 TI - [Long-term results after successful mitral valvuloplasty: comparison of Inoue and double balloon technique]. AB - Mitral balloon valvuloplasty is the treatment of choice for severe mitral stenosis in young patients with a minimally calcified and pliable mitral valve. The present study reports the results of the first 65 patients undergoing mitral valvuloplasty in Zurich with the double-balloon or Inoue-balloon technique. Early outcome and late follow-up over 4.1 +/- 2.5 years were evaluated in these patients. PATIENTS: Percutaneous mitral valvuloplasty was performed in 65 patients (48 females and 12 males, mean age 41 +/- 11 years). The double-balloon technique was used in 25 and the Inoue-balloon technique in 40 patients. Left ventricular pressure as well as pressure gradient and valve area were calculated before and after the intervention. Mitral valvuloplasty was considered to be successful when the valve area was > or = 1.5 cm2 and the pressure gradient < or = 8 mm Hg. RESULTS: Mitral valvuloplasty was successful in 22 patients of group 1 and 39 patients of group 2. Acute complications were observed in 4 patients (6%), i.e. 1 perforation of the left atrium, 1 perforation of the left ventricle, 1 peripheral embolisation and 1 rupture of the mitral leaflet. Mitral valve area increased from 1.0 to 1.9 cm2 with the double-balloon and from 1.0 to 2.0 cm2 with the Inoue-balloon technique. The pressure gradient over the mitral valve dropped significantly from 11 to 4 mm Hg in group 1 and from 15 to 5 mm Hg in group 2. Left ventricular ejection fraction remained unchanged but left atrial pressure decreased significantly in the first group from 20 to 9 mm Hg and in the second group from 22 to 12 mm Hg. Long-term follow-up over 4.1 years showed a mild (not significant) decrease in valve area from 1.7 to 1.6 cm2 in both groups, with NYHA class unchanged and bicycle exercise capacity increased from 76 to 82%. CONCLUSIONS: Mitral valvuloplasty with either the double-balloon or Inoue-balloon technique provides excellent clinical, echocardiographic and haemodynamic results. The long-term follow-up demonstrated a mild decrease in mitral valve area but clinical symptomatology and physical exercise capacity remained unchanged. From a technical standpoint the Inoue-balloon technique is easier to use and has a lower complication rate (2.5%) compared to the double-balloon technique (12%). Thus, in the last few years the double-balloon technique has been replaced by the Inoue-balloon technique, with a good long-term outcome over the first 4-5 years of follow-up. PMID- 11013926 TI - [Controversies in the treatment of carotid stenosis]. PMID- 11013927 TI - [Carotid stenosis: epidemiology and symptomatology (see comment)]. AB - Risk factors for atherosclerotic carotid stenoses are smoking, arterial hypertension, diabetes mellitus and hypercholesterolaemia. Atherosclerotic carotid artery disease is a risk factor for the development of coronary artery disease (CAD), and in older men the risk of CAD-related death is five times higher than the risk of death related to ischaemic stroke. The prevalence of > 50% carotid stenoses in population-based studies in 6-11% for older men and 5-7% in older women. Diagnosis and differential diagnosis of ischaemia-related ophthalmologic and neurological deficits in the territory of the carotid artery are discussed. PMID- 11013928 TI - [Imaging methods in the diagnosis of carotid stenosis: is angiography obsolete?]. AB - The aim of diagnosis is exact grading of carotid stenosis. The existence of different methods of measuring stenosis causes problems. Also, measurement of stenosis using Doppler ultrasound is based on a wholly different principle. Investigative methods for measurement of stenosis are selective angiography, CT angiography, MR angiography and duplex Doppler ultrasound. On the basis of the literature on the advantages and drawbacks, risks and accuracy of the various methods in symptomatic patients, ultrasound is recommended as the primary diagnostic tool. If Doppler ultrasound shows a stenosis of > 70%, MR angiography or CT angiography is recommended. If the results correspond, no further investigation is needed before surgery. If they do not correspond a selective carotid angiogram is required. Sonographic diagnosis of carotid occlusion needs confirmation by MR angiogram or CT angiogram. PMID- 11013930 TI - [Amyloidoma]. PMID- 11013929 TI - [When is interventional therapy necessary for carotid stenosis?]. AB - Stenoses of the internal carotid artery are common. 20-30% of ischaemic cerebrovascular events are due to atherosclerotic lesions of carotid arteries. For many years endarterectomy has been a method for prevention of cerebral ischaemia. There is confusion about how carotid stenosis should be measured on angiograms. Since the results of clinical trials based on different measurement methods are applied to routine clinical practice, the various measurement methods must be comparable. We must know which one was used, and how to convert them by a simple arithmetical equation. Carotid endarterectomy appears to be highly beneficial in patients with recent hemispheric transient ischaemic attacks or non disabling stroke and ipsilateral high-grade stenosis (> or = 60-70%) of the internal carotid artery. Even asymptomatic patients with high-grade stenosis benefit to some extent from surgery, and the beneficial effects correlate directly with the degree of stenosis. Endarterectomy in moderate symptomatic stenosis (50-69%) yields only a moderate reduction in stroke risk. In stenosis < 50% there is no benefit. Decisions about endarterectomy should take into account the natural history of stroke in carotid stenosis, surgical skills, and the individual patient's risk factors. PMID- 11013932 TI - [In Process Citation] PMID- 11013931 TI - [In Process Citation] PMID- 11013933 TI - [In Process Citation] PMID- 11013934 TI - [In Process Citation] PMID- 11013935 TI - [In Process Citation] PMID- 11013936 TI - [In Process Citation] PMID- 11013937 TI - [In Process Citation] PMID- 11013938 TI - [In Process Citation] PMID- 11013939 TI - [In Process Citation] PMID- 11013940 TI - [In Process Citation] PMID- 11013941 TI - [In Process Citation] PMID- 11013943 TI - [In Process Citation] PMID- 11013942 TI - [In Process Citation] PMID- 11013944 TI - [In Process Citation] PMID- 11013945 TI - [In Process Citation] PMID- 11013946 TI - [In Process Citation] PMID- 11013947 TI - [In Process Citation] PMID- 11013948 TI - [In Process Citation] PMID- 11013949 TI - [In Process Citation] PMID- 11013950 TI - [In Process Citation] PMID- 11013951 TI - [In Process Citation] PMID- 11013952 TI - [In Process Citation] PMID- 11013954 TI - [In Process Citation] PMID- 11013953 TI - [In Process Citation] PMID- 11013955 TI - [In Process Citation] PMID- 11013956 TI - [In Process Citation] PMID- 11013958 TI - [In Process Citation] PMID- 11013957 TI - [In Process Citation] PMID- 11013959 TI - [In Process Citation] PMID- 11013960 TI - Distinguishing B and T lymphocytes by scanning electron microscopy. AB - This work demonstrates differences between B lymphocytes and T lymphocytes as seen in the scanning electron microscope (SEM). Slides of routinely prepared benign and malignant lymphoid tissues were viewed in the SEM. The location of prominent populations of T lymphocytes and B lymphocytes was verified by immunocytochemical staining respectively with CD3 and CD20 antisera. Benign tissues, including infant thymus and adult hyperplastic lymph node, were compared with malignant lymphomas of T-cell and B-cell types. The SEM appearance of benign B and T lymphocytes is compared utilizing the backscattered electron (BSE) mode and secondary electron (SE) mode in adult hyperplastic lymph node and infant thymus, respectively. The BSE and SE modes reveal that the sectioned T-lymphocyte nucleus has a more complex configuration than that of the B lymphocyte. T lymphocytes appear more discrete and separated one from another, while B lymphocytes exhibit close cellular association to form a syncytial array. These features are noted also in malignant lymphomas of B-lymphocyte and T-lymphocyte types, respectively. The SEM can distinguish between B and T lymphocytes by studying the differences in nuclear and chiefly the cell to cell appearances. The syncytial configuration of B lymphocytes may reflect prominent expression of cell adhesion molecules, e.g., ICAM-1, as noted in the literature. PMID- 11013961 TI - Scanning electron microscopy of the interior of cells in Hurthle cell tumors. AB - Four cases of Hurthle cell tumor were examined by scanning electron microscopy after being macerated to remove all soluble components. By all morphological criteria, Hurthle cells are oncocytes with their usual augmented complement of mitochondria. The Hurthle cell mitochondria either are ovate with central stacks of cristae or elliptical or rod-like with cristae that often are finger-like. As in salivary gland oncocytes, the shelf-like cristae are anchored to the inner boundary membrane by tubular necks. In some Hurthle cells, all of the mitochondria exhibit reticulate cristae. A few mitochondria harbor a globular inclusion in their inner compartment. The Golgi apparatuses are relatively simple, consisting of imbricated saccules that are edged by small, bud-like structures. The rare lumina in the midst of clusters of Hurthle cells are lined by numerous microvilli. Thus, scanning electron microscopy of macerated Hurthle cell tumors has revealed a number of features, especially of their mitochondria, that have escaped detection by transmission electron microscopy. PMID- 11013962 TI - Clinical, biochemical, and pathological features in a patient with plasma cell dyscrasia and Fanconi syndrome. AB - Multiple myeloma is associated with a wide array of renal diseases that include myeloma cast nephropathy, monoclonal immunoglobulin deposition disease, amyloidosis, cryoglobulinemia, tubular dysfunction, pyelonephritis, nephrocalcinosis, urate nephropathy, and infiltration by atypical plasma cells (or myeloma cells). Filtered immunoglobulin light chains may affect both the distal and, more frequently, the proximal tubule. Tubular abnormalities in patients with plasma cell dyscrasia may be more frequent than previously thought. A patient with a plasma cell dyscrasia is described, who presented with biochemical features consistent with Fanconi syndrome. Immunoelectron microscopy performed on the renal biopsy confirmed the presence of kappa light chain immunoglobulin in intracytoplasmic crystals in proximal tubular epithelial cells. This report is one of a few demonstrating the presence of light-chain immunoglobulin in intratubular crystals in a human renal biopsy obtained from a patient with a plasma cell dyscrasia and Fanconi syndrome. PMID- 11013963 TI - Acinar cell carcinoma of the pancreas. AB - Four cases of acinar cell carcinoma of the pancreas are reported. An acinar cell carcinoma can resemble an islet cell tumor by routine light microscopy but the two differ considerably in their fine structure and immunostaining properties. Although cells of both tumors contain numerous dense-core granules, their size ranges are different, and atypical forms occur in the acinar cell tumors, including elongated bodies filled with filaments. Many mitochondria-rough endoplasmic reticulum complexes were present in one tumor. In a liver metastasis, nests of endocrine cells were discovered amid the groups of acinar cells, and some of the endocrine granules contained rectangular cores. PMID- 11013964 TI - Malignant fibrous histiocytoma: an ultrastructural perspective. AB - Malignant fibrous histiocytoma is a frequent diagnosis, but the relationship of the tumors to histologically similar soft tissue neoplasms is controversial. In this study, 157 examples representing the 4 main subtypes were reviewed by light microscopy and each tumor was studied with the electron microscope. Immunohistochemical stains were performed on 77 tumors. Electron micrographs on 100 fibrosarcomas were reviewed for comparison. Malignant fibrous histiocytomas often closely resemble fibrosarcomas at the ultrastructural level and differences between the two are generally of degree only. Evidence for true histiocytic differentiation was not found. The immunohistochemical results did not contradict the authors' impression from electron microscopy that malignant fibrous histiocytoma forms part of the histologic spectrum of tumors of fibroblasts. PMID- 11013965 TI - The altered structure of renal papillary outflow tracts in obesity. AB - Weight gain is associated with an expanded renal medullary interstitium in humans and in animal models of obesity. In this study, the consequence of obesity and this expanded matrix on renal papillary structure was examined in 15 obese rabbits fed a high fat diet for 8-12 weeks compared to 21 rabbits fed a standard diet. When examined under a dissecting microscope, the tips of the renal papillae from formalin-fixed, methylene blue-stained kidneys showed patent ducts of Bellini in 5 of 5 instances from 2 lean rabbits, but in only 2 out of 12 ducts from 3 obese rabbits. The ostia of the remaining ducts were significantly distended (205 +/- 42 microns versus 56 +/- 8 microns) and occupied by lightly staining granular material. When examined with scanning electron microscopy, all ducts were patent in lean rabbits (6 ducts in 4 rabbits, averaging 104 +/- 12 microns across), whereas only 6 of 11 ducts were patent in papillae from 4 obese rabbits. Renal medullary parenchymal tissue appeared at the openings of the remaining 5 ducts of Bellini in the 4 rabbits. Not only were these 5 ducts significantly distended by the interstitial material (with openings averaging 248 +/- 56 microns across), but the associated collecting ducts were dilated relative to control (100 +/- 15 microns versus 75 +/- 7 microns). Since the ducts of Bellini are the only renal openings that are not corsetted by a fibrous capsule, the authors speculate that the expanded medullary interstitium and increased renal sinus lipid partially obstruct renal outflow and elevate renal interstitial hydrostatic pressure in obesity, causing a prolapse of parenchymal contents, further obstructing urine outflow and leading to distention of the collecting ducts and ducts of Bellini. PMID- 11013966 TI - Cytoplasmic changes in satellite cells of spinal ganglia induced by cisplatin treatment in rats. AB - The effects of cisplatin (cis-DDP) therapeutic treatment on the cytoplasmic compartment of satellite cells (SC) of rat dorsal root ganglia (DRG) were evaluated. Female Wistar rats were treated once a week with i.p. injection of cis DDP (2 mg/kg) for 9 weeks. Morpho-quantitative changes of the cytoplasmatic organelles in SC cytoplasm from L4-L6 DRG were determined at the electron microscopic level. The quantitative changes in the lysosomal system components called dense bodies (DB) and in the mithocondria were stereologically evaluated. The data from SC were compared to those from the neurons. The cis-DDP treatment induced a great increase in DB and mithocondria volume of SC. Furthermore, the SC sheath showed an increase of the cytoplasmic lamellar expansions responsible of the physical dissociation of SC sheath from the nerve cell body surface. The comparative analysis from SC and neurons showed that the drug affected primarily the SC, supporting the idea that SC could be the initial target of cis-DDP molecule. The alterations of the anatomical relationships between SC and neurons could modify the cell control on extracellular solutions, altering the functional role barrier attributed to SC. It appears that not only the DRG neurons but also and principally the SC were involved in the peripheral neuropathy mechanisms caused typically from therapeutic cis-DDP administration. PMID- 11013967 TI - Meningioma with granulofilamentous inclusions. AB - The authors report a case of intracranial meningioma with granulofilamentous inclusions. A 50-year-old man had right trigeminal neuralgia due to trigeminal nerve compression by a petroclival tumor and received tumor resection. Microscopically, tumor cells containing eccentric nuclei and intracytoplasmic hyaline inclusions were arranged in sheets and whorls. The inclusions were negative for periodic acid-Schiff reaction. No histological anaplasia was seen. Immunohistochemistry showed epithelial membrane antigen reactivity on the cytoplasmic membrane. Immunoreactivity for vimentin was recognized in cytoplasm adjacent to inclusions. However, confocal laser microscopic study revealed immunoreactivity for vimentin even inside some inclusions. Ultrastructurally, interdigitation of cytoplasmic processes and desmosomes connecting adjacent cells were noted. Inclusions were composed of numerous fine osmiophilic granules attached by intermediates filaments. These findings were consistent with a meningioma with the granulofilamentous inclusions described earlier. The findings demonstrated by confocal laser microscopy and electron microscopy suggest that these granular materials may be the metabolic products of vimentin filaments. PMID- 11013968 TI - Apparent progression of acute glomerulonephritis to dense deposit disease. AB - One week after the diagnosis of meningococcal meningitis, an 8-year-old boy presented with acute renal failure and hypocomplementemia. A renal biopsy showed "postinfectious glomerulonephritis" and acute tubular necrosis. Hematuria, proteinuria, and low complement levels persisted, and 2 years later a follow-up renal biopsy revealed dense deposit disease. The apparent progression of postinfectious glomerulonephritis to dense deposit disease as observed in this patient has not been previously described. PMID- 11013969 TI - Cardiology and cost control: the ethical challenge for the new millennium. AB - Government interventions in the health care sector threaten the traditional role of physicians, since they are increasingly forced to consider the cost of medical care when making decisions on behalf of their patients. To prepare themselves for this ethical challenge and to actively participate in the debate about cost containment, physicians need to understand how health economists and politicians view the problem of rising medical costs. This review summarizes some essential facts and findings of the health economics literature that provide the rationale for different approaches to cost containment. The effects of rapidly growing health care cost on the economy are discussed, and improvement of medical technology is identified as the driving force behind this growth. The different policy instruments, which can be employed for cost containment, are explained against this background with an emphasis on Managed Care and global budgets. The outlined concepts are finally discussed in the context of the current debate about the proposed cost containment legislation in Germany. PMID- 11013970 TI - [C-reactive protein as an independent marker of prognosis in acute coronary syndrome: comparison with troponin T]. AB - BACKGROUND: It has been suggested that inflammatory processes play a role in the pathogenesis of acute coronary syndromes (ACS). C-reactive protein (CRP) is a classic acute phase protein. It is yet unclear whether, in addition to established markers as troponin T (TnT), determination of CRP in patients admitted for ACS contributes significantly to the diagnosis and prognosis of ACS. PATIENTS AND METHODS: We investigated 50 patients with ACS (59.4 SD 13.9 years) in the first hour after admission and 4-24 h later with respect to TnT (Elecsys, Roche Diagnostics) and CRP (biokit, modified Quantex CRP plus, analytical sensitivity 0.02 mg/dL). Fifty percent of the patients were classified as having unstable angina retrospectively. All patients were followed in the 6 weeks post discharge regarding death and recurrent ACS. RESULTS: The cumulative event rate at 6 weeks after discharge was 62.5% for patients being CRP and TnT positive compared to 35.3% in TnT positive and CRP negative patients. In TnT negative patients a positive CRP test predicted 33.3% of events and 28.8% of patients negative for CRP and TnT had events at 42 days post discharge. Logistic regression analysis regarding the primary endpoint including TnT and CRP (4-24 h values), age, gender and diagnosis resulted in independent prediction of ACS or death by TnT (cutoff 0.1 microgram/L, p = 0.048, odds ratio = 7.5) and CRP (cutoff 0.862 mg/dL, p = 0.026, odds ratio = 5.3). Sensitivity/specificity for AMI diagnosis were 69.6%/75% for TnT and 12%/72% for CRP in the first hour and 91.3%/68.2% for TnT and 68%/72% for CRP 4-24 h later. CONCLUSIONS: Besides TnT, high sensitivity CRP determination has no additional value for early AMI diagnosis. The prognosis of these patients during the first 24 hours is significantly and independently predicted by CRP measurements in addition to troponin T. PMID- 11013971 TI - [Risk stratification in heart surgery: do risk scores facilitate the determination of indications?]. AB - Various risk scores have been developed for the assessment of operative risk in cardiac surgery. Although risk stratification has been acknowledged as a useful tool to analyze trends in therapy and changes in patient populations, its relevance in assessing the indication for surgery has been questioned. It was the goal of this prospective study to compare 6 common risk scores with regard to the predictive value for mortality in individual patients. Between September 1998 and February 1999 all adult patients undergoing heart surgery were prospectively scored according to the following scores: initial Parsonnet, Cleveland Clinic, French, Euro, Pons, and the Ontario Province Risk score. Early lethality was assessed within 30 days postoperatively. Follow-up was completed in 504 patients. With the exception of the Ontario Province Risk score, lethality in the high risk group was overestimated by all scores, whereas lethality in low to moderate risk groups was underestimated by several scores. Mean scores of surviving and deceased patients showed a broad overlap with high standard deviations. Preoperative risk scores are effective tools for stratification of patient populations and the analysis of surgical outcome. With the aid of risk scores, operative risk can be sufficiently predicted for patient populations or subpopulations. The Euro score best predicted the outcome of our patients. However, when the indication for surgery is to be determined in an individual patient, risk scores should be only considered as an orientation in the decision process. PMID- 11013972 TI - [Restenosis predictors after stent implantation of venous aortocoronary bypass grafts]. AB - Follow-up studies after stent implantation of native coronary arteries have reported reduced rates of angiographic restenosis. In contrast, stent implantation in the treatment of obstructive disease of coronary artery bypass grafts is complicated by higher restenosis rates. We sought to determine, if different predictors contribute to the high restenosis rate following stent implantation of coronary artery bypass grafts. We investigated long-term angiographic outcome of 205 stent implantations performed in 177 patients. Multivariate analysis correlated clinical, procedural and angiographic variables with the incidence of angiographic restenosis defined as diameter stenosis > 50% at follow-up. Angiographic restenosis was observed in 34% of lesions treated. Multiple logistic regression analysis defined diabetes mellitus (OR 6.89, CI 2.41 9.69), graft recanalization (OR 2.69, CI 1.08-6.63), lesion at the aortic anastomosis (OR 6.98, CI 2.76-19.25), lesion at the coronary anastomosis (OR 2.95, CI 1.18-7.49), high diameter stenosis after stent placement (OR 7.01, CI 2.64-15.71), placement of long stents (OR 2.78, CI 1.11-7.36) and implantation of more than one stent (OR 7.34, CI 2.08-20.15) as independent predictors of graft in-stent restenosis. Critical consideration of these variables may help to identify patients who are poor candidates for stent implantation and who may benefit from different interventional approaches. PMID- 11013973 TI - [Is Na+Ca(2+) exchanger expression altered in the endomyocardium of patients with chronic heart valve diseases parallel to myocardial dysfunction?]. AB - BACKGROUND: Na(+)-Ca2+ exchanger (EXCH) is an important regulator of intracellular calcium homeostasis. To maintain a normal intracellular Ca2+ concentration, EXCH expression may be upregulated before the onset of end-stage heart failure. We tested for a correlation between the EXCH transcription level and the degree of myocardial dysfunction as well as the suitability of EXCH transcription as a molecular marker for early detection of a transition from adequate to inadequate myocardial adaptation to chronic pressure and/or volume overload in valvular heart disease (VHD). METHODS: The level of EXCH transcription was analyzed in myocardial biopsies from eleven patients with aortic stenosis (AS), five with aortic regurgitation (AR) and six with primary mitral regurgitation (MR) of different hemodynamic severity and myocardial impairment using the quantitative rt-PCR technique. In addition, endomyocardial tissue from thirteen explanted hearts with end-stage heart failure and biopsies from seven individuals without heart disease were investigated. RESULTS: The mean level of EXCH transcription in patients with AS was: 1.8 +/- 1.4 amol/ng total RNA, with AR: 1.9 +/- 0.8 amol/ng and with MR: 2.2 +/- +2.1 amol/ng. This was not from different controls (2.6 +/- 1.2 amol/ng total RNA). However, in myocardium from end-stage heart failure, EXCH transcription was increased fourfold amounting to 8.9 +/- 1.9 amol/ng total RNA. No difference in the EXCH transcription was found in VHD with respect to the degree of myocardial dysfunction: cardiac index (CI) > 3.5 l/min/m2 (EXCH 1.4 +/- 1.1 amol/ng total RNA); CI 3.5-2.4 (EXCH 2.5 +/ 1.8); CI < 2.4 (EXCH 1.8 +/- 1.0); EF-angio > 50% (EXCH 1.9 +/- 1.8); EF-angio < or = 50% (EXCH 1.9 +/- 0.9); EF-RNV > 50% (EXCH 2.4 +/- 1.8), EF-RNV < or = 50% (EXCH 1.7 +/- 1.0). CONCLUSION: Myocardial EXCH transcription does not change parallel to the degree of myocardial dysfunction in VHD. Consequently, myocardial EXCH transcription does not appear to be suitable as a parameter indicating the transition from adequate to inadequate myocardial adaptation to chronic volume and/or pressure overload. PMID- 11013974 TI - [Impediment of cellular immune response under treatment with ticlopidine in a patient with Staphylococcus aureus endocarditis]. AB - A 52-year-old male with coronary artery disease was admitted with acute aortic valve endocarditis and a temperature up to 39.5 degrees C caused by Staphylococcus aureus. The patient was treated with ticlopidine (Tiklyd) after percutaneous transluminal coronary angioplasties to reduce restenosis by inhibiting thrombocyte aggregation. Upon admission c-reactive protein (CRP) was 389 mg/l. Interleukin-6 (IL-6) and Interleukin-2-receptor (IL-2-rec) were distinctly increased. Monoclonal antimyocardial antibodies were found. Leukocyte count never exceeded 9.8 G/l; however, transesophageal echocardiography validated a soft vegetation of the aortic valve. Antibiotic therapy was initiated with imipenem, gentamicin and vancomycin; clarithromycin was added after five days. Temperature normalized after 24 days. The c-reactive protein decreased from 389 mg/l to 6 mg/l, and the elevated cytokine levels decreased accordingly. Agranulocytosis or pancytopenia by ticlopidine through a toxic mechanism have been described, which are normally reversible within three weeks; there has not yet been a description of a missing leukocyte response in endocarditis as in this case report. This is a special situation with lack of or impeded immunological response, which limits the use of ticlopidine, especially since a therapeutic alternative with clopidogrel is available. PMID- 11013975 TI - [Fibrinolytic therapy in thrombosis of a mitral valve prosthesis]. AB - A 48-year-old woman presented with progressive dyspnea due to thrombosis of a mitral valve prosthesis. The patient had undergone mitral valve replacement (St. Jude Medical) six years prior to admission because of mitral stenosis (Class III); three years later the prosthesis had to be replaced (St. Jude Medical) because of valve thrombosis. At admission, transesophageal echocardiography showed a thrombus on the atrial side of the fixed valve leaflet and a thrombus (2.4 x 1.6 cm) floating from the left atrial roof. Because of the previous thoracotomies, thrombolysis was initiated despite the mobile thrombus with the attendant risk of embolization. Urokinase was infused in a dose to maintain the fibrinogen level around 100 mg/dl. After 24 h, the mean pressure gradient across the prosthetic mitral valve (measured by doppler echocardiography) had decreased from 23 to 11 mmHg. After 13 days of this modified thrombolytic regimen, the clinical symptoms of the patient had resolved and echocardiography showed a normal function of the prosthetic mitral valve without evidence of residual thrombosis. This patient demonstrates that prolonged cautious thrombolysis can be effective for the treatment of prosthetic valve thrombosis in hemodynamically moderately compromised patients. PMID- 11013976 TI - [Ventricular septal aneurysm in a patient with Marfan syndrome]. AB - This is the first description of a patient with Marfan syndrome and an aneurysm of the ventricular septum. Apart from a borderline dilatation of the ascending aorta, there were no cardiovascular manifestations of Marfan syndrome. A transesophageal echocardiographic examination showed a large aneurysm of the ventricular septum. To prevent the imminent rupture and the acute occurrence of a significant left-to-right-shunt on the ventricular level, as well as increasing irritation of tricuspid valve, an elective operation was performed. Besides the routine cardiological diagnostics, the search for intracardial defects is also necessary in patients with Marfan syndrome. PMID- 11013977 TI - [Recommendations of the International Cardiology Forum (ICF) 1998 on revision of the current guidelines for the diagnosis and treatment of unstable angina pectoris and nontransmural myocardial infarct]. AB - Rapid progress in the understanding and treatment of unstable angina and acute coronary syndrome are prompting occasional revisions to current treatment guidelines. The recommendations contained in this paper are based on the consensus reached during discussions at the 'International Cardiology Forum' in September 1998. Consensus was reached on significant major points, although some aspects remain controversial. A substantial body of data accumulated in a host of studies justify changes to current treatment habits. PMID- 11013978 TI - [Use of Abciximab in threatening vascular occlusion after PTCA]. AB - The administration of GP IIb/IIIa antagonists has been shown to be effective in reducing myocardial infarction and cardial death when given before PTCA. This prospective study was performed to determine the efficacy of abciximab in a bail out situation to manage threatened or acute vessel closure. METHODS: Acute or threatened vessel closure was observed in 104 (5.5%) out of 1903 consecutive patients treated with PTCA in our institution. Of the 104 patients 46 (44%) were treated for unstable angina (CCS IV). Abciximab was administered in bail-out situations in a dosage of 0.25 mg/kg given as a bolus, which was followed by an intravenous infusion of 10 micrograms/min over 12 hours. Repeat PTCA was performed shortly after the administration of the abciximab bolus. After the procedure, the sheath was left in place and control angiography was carried out 24 h later. RESULTS: In 100 of the 104 patients TIMI flow III could be restored by abciximab therapy and RePTCA. In 4 patients an additional stent implantation was necessary due to persistent flow limitation. One day post PTCA, early follow up angiography demonstrated patency of all vessels except two. In-hospital events occurred in 4 patients. Three of these patients underwent emergency CABG due to subacute vessel closure a few hours after PTCA and died during or directly after surgery. Follow-up after one year included clinical status and control angiography of the target vessel. During long-term follow-up, MACE occurred in 15 patients (2 MI, 8 CABG and 5 RePTCA). CONCLUSION: The results of this prospective trial demonstrate the efficacy of abciximab therapy in bail-out situations occurring during or early after PTCA. The use of abciximab in bail-out situations appears clinically beneficial. Further studies have to compare the efficacy of this approach with prophylactic abciximab treatment. PMID- 11013979 TI - [Ross operation in aortic valve diseases]. AB - Currently used standard mechanical and bioprosthetic heart valves all have limitations that produce less than optimal results. The pulmonary autograft offers a promising alternative with theoretical advantages that have to be proven for final judgement of this innovative surgical technique. METHODS AND RESULTS: A survey including the international registry with 2523 patients enrolled since 1987 is provided along with the author's 10 years experience of 157 patients. The age of the patients ranges between one and 79 years. The indication is expanding integrating concomitant procedures like repair or replacement of aneurysms of the aortic root and ascending aorta, mitral valve reconstruction, coronary bypass grafting, etc. The operative mortality ranges between 0.6 and 2.5%. No anticoagulation is necessary. The function of the autograft is excellent. Late incidence of significant aortic insufficiency (> 2+) remains less than 5%. Even at exercise there is no pressure gradient across the autograft. Autograft failure with autograft revision or replacement stands at 1-4%. Homograft function is also found to be excellent. Right ventricular outflow tract revision rate ranges between 1-1.3%. In a few patients (7%), maximal pressure gradients between 20 and 35 mmHg developed across the homograft without major impairment of right ventricular function. CONCLUSIONS: The multicenter data confirm the superior performance of the autograft not matched by any other biological or mechanical valve replacement. Scientific efforts are required to further improve long-term function especially of the neopulmonary valve. PMID- 11013980 TI - ["Vacation, again...?"]. PMID- 11013981 TI - [Responsibility of the rheumatologist in skeletal radiology. A contribution to the "Bone and Joint Decade"]. PMID- 11013982 TI - [Patient education in rheumatology--quality control]. PMID- 11013983 TI - [Spinal stiffness. Differential imaging diagnosis with reference to acquired hyperostosis syndrome and spinal loading disease]. AB - There are numerous diseases which can irreversibly stiffen the whole spine or parts of the axial skeleton. Due to didactic reasons one can distinguish 3 groups of such disorders: Paradigmatic diseases of the spine with an inherent proneness to stiffness. Rare diseases which involve a certain danger of stiffening the spine. Rigidity of the spine due to scientifically unrecognised morphologic phenomena. PMID- 11013984 TI - [Possibilities and limits of conservative therapy of spondylitis and spondylodiscitis]. AB - BACKGROUND AND AIM OF THE STUDY: For many years the treatment of spondylitis and spondylodiscitis has been discussed controversially. The aim of this study is to report on objective and subjective mid-term results of therapy of spondylitis and to present a differentiated concept of treatment. METHODS: Between 1988 and 1996, 58 patients were treated with spondylitis or spondylodiscitis. Nine of these patients had to be operated. A biopsy was taken in all of the cases. According to the antibiogram obtained by the biopsy, antibiotics were applied intravenously. The patients were immobilized by a plaster bed for at least 6 weeks and were then treated by a spinal orthosis for another 3 months. Patients were re-examined clinically and radiographically and by a questionnaire (including Roland-Morris score), after 8 years on average. RESULTS: Patients were diagnosed correctly 4 months after the begin of the disease. Spondylitis was predominantly localized in the lumbar spine. A positive bacteriological culture was derived from one third of the biopsies; none of them was a specific culture. C-reactive protein was revealed as appropriate for diagnosis and follow-up of spondylitis. In 84% of the patients a total or partial bony fusion was demonstrated radiographically. Questionnaire assessment revealed a significant decrease of the Roland-Morris score (17.8-7.4) and a significant relief of pain (8.9-2.5). CONCLUSIONS: We recommend surgical treatment on patients with major vertebral body destruction, epidural abscess and progressive neurological impairment. These indications for operation can be prevented by a rapid diagnosis, so that spondylitis can be successfully treated by consequent immobilization in a plaster bed and appropriate antibiotics. PMID- 11013986 TI - Network Symposium Rheumatology and Clinical Immunology. March 23-25, 2000, Berlin, Germany. Proceedings and abstracts. PMID- 11013985 TI - [Mixed tophi. Calcium pyrophosphate dihydrate crystals in gout tophi]. AB - Calcification of tophi is a well-known secondary phenomenon in gout. Because there are no data available on the nature of the calcium component, light and scanning electron microscopic as well as X-ray microanalysis studies were performed on calcified tophi. In unstained histological slides, urate crystals were detected as negative birefringent needles; after incubation of the slides in distilled water, the urate crystals had disappeared, only positive birefringent calcium pyrophosphate dihydrate (CPPD) crystals of different size remained. An identical result was obtained by scanning electron microscopy. By X-ray microanalysis, peaks for calcium and phosphorus were measured, indicating the presence of CPPD. The occurrence of CPPD crystals in urate tophi as well as in foreign body granulomata indicates that the occurrence of CPPD is not restricted to articular tissues. The biochemical mechanism leading to the appearance of these crystals remains unknown, but it may be assumed that an increase of connective tissue destruction may be responsible for this calcification process. PMID- 11013987 TI - [Quality standards of the German Society of Rheumatology for Rheumatologic Patient Education--preliminary report, discharged by the governing body and with consultation of the German Society of Rheumatology in September 1999]. PMID- 11013988 TI - [Recommendations of the German Society of Rheumatology for therapy with tumor necrosis factor inhibitors. Pharmacotherapy Committee of the German Society of Rheumatology]. PMID- 11013989 TI - [Anesthesiological care in removal of bulky formations from functionally important hemispheric zones: craniotomy in conscious state]. AB - A method of anesthesia allowing patient's awakening to the level of verbal contact after trephination and opening of the dura mater was used in 37 patients with bulky formations (33 patients with tumors and 4 with arteriovenous malformations) in the speech zones of speech-dominant hemisphere. Speech mapping of the brain, carried out in alert patients, helped eliminate permanent speech deficiency during the postoperative period in all patients. The protocol of anesthesia was as follows: locoregional anesthesia of soft tissues of the head with a mixture of 2% xylocaine and 0.5% marcaine with epinephrine intravenous diprivan at stages requiring no patient's alertness, clopheline, fractionated midazolam (after identification of speech zones). The study showed that this protocol helps solve the problem and adequately protects the organism from surgical stress, provides good conditions for manipulations on the brain, involves no unpleasant sensations for the patient, and in 70% patients helps attain complete amnesia of the perioperative period. The most severe complication is an epileptic attack. Respiratory monitoring (pulse oximetry and capnography in the lateral duct) is obligatory for preventing respiratory disorders in patients on spontaneous respiration who were not intubated. PMID- 11013990 TI - [Cerebral oximetry in neurosurgical patients with cerebrovascular diseases. I. Analysis of causes of intraoperative changes in rSO2 values and its prognostic significance]. AB - A total of 144 neurosurgical interventions were performed in 138 patients with cerebrovascular diseases (arterial aneurysms in the acute and cold periods after a subarachnoidal hemorrhage, arteriovenous malformations, and carotid-cavernous anastomoses). Intraoperative monitoring of regional saturation of hemoglobin with oxygen in the blood flowing in brain vessels was carried out by a cerebral oximeter INVOS 3100 for early intraoperative diagnosis of cerebral ischemia. This paper analyzes factors affecting changes in rSO2 parameter during surgery and discusses the prognostic significance of the detected shifts. Various factors affected the time course of rSO2: decrease in AP within the framework of controlled arterial hypotone and other causes, insertion of autosupported retractor spatulae, temporary clipping and embolism of cerebral arteries, vasospasm, aneurysm rupture, dissection of arteriovenous malformation, etc. Any intraoperative shift of rSO2 surpassing 5%, no matter of what direction (decrease or increase), deserves special attention of anesthesiologist, as it indicates development of cerebral ischemia with a high degree of probability. PMID- 11013991 TI - [Cerebral oximetry in neurosurgical patients with cerebrovascular diseases. II. Comparison of electrophysiological methods of examination and cerebral oximetry]. AB - Simultaneous monitoring by cerebral oximetry and electrophysiological methods (electroencephalography and electrocorticography) was performed in 21 patients with arterial cerebral aneurysms during direct intracranial interventions, endovascular interventions, and Matas' test. Electrophysiological methods are sufficiently accurate indicators of cerebral ischemia even at a functionally reversible level. The results of simultaneous monitoring were similar in 80% cases, indicating cerebral ischemia even at functionally reversible level, which was detected by both electrophysiological methods and cerebral oximetry. Variously directed shifts were detected in 4 cases; the causes are discussed. Cerebral oximetry detects cerebral ischemia with a sensitivity similar to that of electrophysiological methods. The method does not depend on temperature or anesthetics. Limitations of the method are due to its local application. PMID- 11013992 TI - [Pharmacological protection of the brain during surgery in patients with giant cerebral aneurysms]. AB - The authors discuss the results of pharmacological protection of the brain, consisting in intraarterial or intravenous infusion of drugs (sodium thiopental, nimodipine, ketamine) during intracranial interventions in 13 patients with giant arterial aneurysms of the anterior portions of Willis' circle. In some patients drug protection of the brain prevented the development of ischemic focus in the basin of temporarily occluded vessel. Intraarterial infusion of protector drugs is preferable because it allows decreasing the doses and severity of untoward hemodynamic effects. PMID- 11013993 TI - [Epidural anesthesia in surgical interventions on the spine and spinal cord. I. Comparative analysis of the effectiveness of anesthesiological protection under conditions of epidural anesthesia and neuroleptanalgesia in surgery of the spine and and spinal cord]. AB - The study was carried out in 22 patients operated on for vertebral disk hernias and spinal tumors at lumbosacral level. The patients were divided in 2 groups depending on the type of anesthesia (epidural or neuroleptanalgesia-EA and NLA). In the test group all patients were operated under EA with local anesthetics combined with intravenous sedative drugs (diprivan + relanium) under conditions of spontaneous respiration and O2 inhalation through a mask. In the control group combined total intravenous anesthesia by myorelaxants and tracheal intubation were carried out (relanium + diprivan: induction dose 1.95 +/- 0.5 mg/kg, maintenance dose 5.3 +/- 0.4 mg/kg/h, and phentanyl). The purpose of the study was to compare the efficiency of anesthesiological protection under EA and traditional NLA in interventions on the spine, when surgical injury is inflicted in the immediate vicinity to the central structures responsible for pain impulsation. Only EA ensured adequate protection of the patients from surgical stress, as was seen from hemodynamic (arterial pressure and heart rate) and endocrine metabolic parameters (glucose, epinephrine, norepinephrine, hydrocortisone, and prolactin levels). Hence, EA fully demonstrated its protective properties during operations on the spine, and therefore can be regarded as a method of choice in this patient population. PMID- 11013994 TI - [Epidural anesthesia in surgical interventions on the spine and spinal cord. II. Effects of epidural anesthesia on somatosensory evoked potentials]. AB - Effects of epidural anesthesia (EA) on early components of somatosensory evoked potentials (SSEP) were studied and the objectiveness and efficiency of SSEP monitoring during interventions on the spine under EA were evaluated. Evoked potentials were studied in 21 patients operated on for vertebral disk hernias and extra-intradural tumors of the spine (lower thoracic and lumbar levels) under bupivacaine EA. Cortical SSEP were recorded and analyzed on a Viking IV neuroaverager (Nicolet, USA) stimulating n. tibialis posterior for obtaining a greater amplitude of evoked potentials. The following SSEP characteristics were measured: latency of PI (P37), NI (N45), and PII (P60) peaks, amplitude of PINI peak, and inter-peak latency of PI-NI and PI-PII. The major changes in cortical SSEP caused by EA (local anesthetic 0.5% bupivacaine) involve only PI and NI components which reflect the entry of information on an external stimulus into the cortex and objective physical parameters of this stimulus; this helps predict the onset of full-value epidural block and its duration. General anesthetic (propofol) affect mainly a later component of response, PII peak, which is responsible for processing of primary information about an external stimulus and reflects the activation of associative areas of the brain. Hence, SSEP regulation can be used together with traditional methods (pin prick test and Bromage scale) for evaluating epidural block in patients with spinal diseases. PMID- 11013995 TI - [Early awakening or prolonged sedation after neuroanesthesia: which is better for the patient?]. PMID- 11013996 TI - [Anesthesia in magnetic resonance tomography]. AB - Sixty patients were anesthesized for MRI. Intravenous and/or intramuscular anesthesia with ketamine, diazepam, pipolphen (promethazine) was supplemented by aminazine (chlorpromazine) or tisercin for adult patients. Spontaneous respiration was maintained, which helped avoid problems with endotracheal intubation and assisted ventilation. The method is safe, economic, and its side effects are negligible. Main features of MRI and specific anesthesiological problems associated with this method are discussed. PMID- 11013998 TI - [Effects of various protocols of respiratory support on hemodynamics in neurosurgical patients]. AB - Central hemodynamics is analyzed in neurosurgical patients during application of various methods of assisted ventilation. Invasive monitoring of central hemodynamics was performed during respiratory support of neurosurgical patients for the first time in Russia. Gradual decrease of PSV helps evaluate the readiness of patients to decrease of respiratory support. Metabolic respiratory oxygen value at least 37%, cardiac index at least 4.5 +/- 0.5 liter/min x cm2, and working load of the left and right heart at least 6.7 +/- 0.75 and 0.71 +/- 0.183 kg/m(min x m2), respectively, are hemodynamic criteria for transferring to spontaneous respiration. PMID- 11013997 TI - [Basic intensive care of ischemic cerebral stroke. Method of therapeutic hemodilution]. AB - Ischemic cerebral stroke (ICS) ranks among the most frequent causes of disability. The incidence of untoward consequences of ICS can be decreased by early adequate use of intensive care methods restoring tissue oxygenation in the focus. The most important methods of intensive care of ICS are measures notably improving microcirculation and essentially decreasing subsequent incidence of disabling complications. The basic component of intensive care ensuring favorable results is therapeutic hemodilution making use of modern plasma substitutes- second-generation hydroxyethyl starch Infukoll HES 6 and 10% solutions. Currently used methods including use of dextran-based solutions cannot normalize microcirculation in ICS patients, particularly if used in long course. Prolonged infusions of dextran-based solutions inevitably increases plasma viscosity and impairs the hemostasis system. An essential increase of plasma viscosity involves the need in a complex of appropriate drug therapy aimed at compensation for untoward aftereffects of high dextran doses. Therapeutic doses of dextran-based solutions do not notably improve blood rheology. By contrast, therapeutic hemodilution with Infukoll HES notably improves the results of intensive care of ICS. PMID- 11013999 TI - [Nosocomial pneumonia in patients with severe craniocerebral trauma in intensive care units]. AB - Eighty-three patients with severe craniocerebral injuries (CCI) were treated at Institute of Neurosurgery in 1999. Pulmonary infectious complications occurred in 16 of 25 patients with severe CCI. Early nosocomial pneumonia (NP) was diagnosed in 18% and the so-called late NP (associated with artificial ventilation of the lungs) in 35%. Coma longer than 4 days increased the incidence of NP to 62%. The main pathogens of NP are gram-negative aerobic bacteria (61%), the predominant agent being Pseudomonas aeruginosa (18.9%). 76% isolated microorganisms were multiresistant. The most significant risk factors as regards NP in patients with severe CCI were coma combined with bulbar and pseudobulbar disorders. PMID- 11014000 TI - [Principles of intensive care during the acute period of severe craniocerebral trauma in children]. AB - Prospective analysis of the course of severe craniocerebral injury (CCI) in 37 children showed that the results of treatment depended on methods of intensive care. In children the outcome of CCI depends primarily on adequacy of maintaining effective cerebral bloodflow during the acute period of CCI irrespective of the terms and etiological factors of coma. Adequate cerebral perfusion pressure is attained with osmodiuretics, saluretics, and hemodynamic therapy. Hyperventilation is justified in cases with coma developing within 2 h after the injury; it should not lead to long (more than 1 h) decrease of PaCO2 below 28 mm Hg. The authors emphasize that every delayed decrease of consciousness after the injury should be thoroughly analyzed in order to rule out intracranial complications (epi/subdural and intracerebrai hematomas) in no more than 6 h, which is the maximal time allowed for decision making. Use of unapproved medical methods and drugs in children during acute period of severe CCI is ineffective and even hazardous, because it distracts the physician's attention to negligible details of treatment; in addition many of such drugs have side effects. PMID- 11014001 TI - [Respiratory failure in disseminated sclerosis]. AB - The development and patterns of respiratory failure (RF) are analyzed in 9 patients with disseminated sclerosis (DS). Forced ventilation of the lungs was carried out with consideration for main location of the process. Relationship between patterns of respiratory disorders and neuroanatomy of respiratory regulation is discussed. Involvement of the corticospinal routes is paralleled by dissociation during functional pulmonary tests: spontaneous volumes are less than controlled inspirations. The most severe symptom complexes were observed in RF of predominantly bulbar localization: respiratory anarchy, blocking of airways caused by impaired swallowing, impaired mechanism of coughing reflex, loss of spontaneous respiration, sometimes apnea during sleeping. Involvement of the respiratory nuclei of medullary respiratory center and airways and of the corticonuclear routes of caudal cranial nerves causes the development of a triad of symptoms: glossopharyngolaryngeal paralysis, dysfunction of respiratory nuclei of medulla oblongata, and decreased sensitivity of respiratory center to CO2. Aspiration complications caused by dysphagia are characteristic of bulbar DS. Respiratory function in 5 patients without clinical picture of RF are specially discussed. The authors emphasize unfavorable prognostic significance of signs of extracorporeal obstruction indicating the probability of RF long before its manifestation. Special attention is paid to early diagnosis of symptoms of coming RF when evaluating the status of patients with DS during treatment. Timely use of respiratory resuscitation methods reduces the mortality and ensures a good chance for remissions with recovery of respiratory function, which are characteristic of RF. PMID- 11014002 TI - [Videofluoroscopy study of swallowing in neurogenic dysphagia]. AB - Videofluoroscopy (VFS) was for the first time used for examining swallowing in 49 patients with nervous diseases. Disturbances in each phase of swallowing act are analyzed with evaluation of the time parameters and defects, causes of aspiration in neurogenic dysphagia are discussed, and cricopharyngeal insufficiency is described. Neurogenic dysphagia is characterized by a combination of disorders which determine the degree of dysphagia. The most severe swallowing disorders were observed in patients with multiple foci in the brain stem and in diphtheritic polyneuropathies. The authors conclude that VFS is the optimal method for the diagnosis of neurogenic dysphagia. PMID- 11014003 TI - [Prevention of postoperative venous thromboembolic complications]. AB - Venous thromboembolic complications (VTEC) occurring during the postoperative period notably increase the mortality. Prevention of this complications requires detection of risk factors in order to evaluate the risk for every patient. Based on the risk factors described in this paper, all patients can be divided into categories at a high, moderate, and low risk of complications. Preventive measures corresponding to the risk are to be taken. These measures are to include measures improving venous flow and anticoagulant therapy (low-molecular heparins should be preferred). Though these measures do not 100% prevent thromboembolic complications, they notably decrease the risk of VTEC. PMID- 11014004 TI - [New long-acting local anesthetic: ropivacaine hydrochloride (naropin)]. PMID- 11014005 TI - [Case of uncommon motor reaction to administration of diprivan]. AB - A young patient without history of epilepsy developed a pronounced motor reaction simulating an epileptic attack in response to diprivan. Possible causes of motor excitation in response to diprivan and probability of their prediction are discussed. PMID- 11014006 TI - ["Artificial liver"]. AB - Acute liver failure is a life-threatening condition since conventional medical treatments have little effect on survival. Artificial liver support systems based on blood detoxification alone have proven to be ineffective. A liver support system should carry out essential functions such as the phase I reaction in which lipid-soluble toxic substance are rendered water-soluble by the enzyme system of the cytochrome P450 and NADPH-cytochrome reductase, and are therefore conjugated by the phase II reaction, before excretion. Liver support systems should be capable of sustaining patients until an organ is available for liver transplantation (bridging treatment), or improving the survival in patients for whom liver transplantation is not a therapeutic option. Recent advances in cell biology and tissue culture techniques have led the way for potential clinical use of isolated hepatocytes so that they are now an important element of bioartificial liver (BAL) support devices. Some of these BAL are currently under clinical investigation in the USA and Europe, and the results of the prospective controlled trials will be soon available. PMID- 11014007 TI - [Fulminant liver failure: etiopathogenesis and therapy]. AB - Fulminant hepatic failure (FHF) is a clinical syndrome characterized by the development of encephalopathy within eight weeks from the onset of the first symptoms, in the absence of previous hepatic disease. It is an uncommon but not rare disease, often fatal but potentially reversible. This article looks at the diverse aetiologies, clinical features, and current medical management, including orthotopic liver transplantation, and auxiliary orthotopic or eterotopic liver transplantation, that are the most recently adopted surgical procedures. Clinical experience with bio-artificial liver support systems of two of the most active research Groups in this field, concludes the paper. PMID- 11014008 TI - [Acute and fulminant liver failure: experimental models]. AB - Fulminant hepatic failure (FHF) is a complex clinical syndrome, with an invariably high mortality rate, that follows many possible and different infectious, pharmacologic and surgical liver injuries. The appearance of the syndrome is similar whatever the etiology, but the mechanisms which lead to the development of FHF are greatly varied. In order to understand the possible pathways which drive to FHF, experimental animal models have been used for a long time. Six requirements should be fulfilled by any FHF animal model: 1) reversibility; 2) reproducibility; 3) death from liver failure; 4) the presence of a therapeutic window; 5) the need of large laboratory animal; 6) minimal hazard to personnel involved in the study. In the present paper a number of models are reported and described, and advantages and disadvantages are discussed. It is concluded that with respect to the aforementioned criteria, no available experimental model is yet as satisfactory as expected. PMID- 11014009 TI - [Acute liver failure in chronic hepatic disease. Clinico-therapeutic evaluation]. AB - Chronic liver diseases are potentially evolving clinical situations which, independently by the etiology, could proceed towards progressive liver structural and functional impairments. The only efficient treatment is orthotopic liver transplantation. Chronic liver diseases, and up to 40% of liver cirrhosis, are initially asymptomatic, but cirrhosis is the most frequent cause of death among non-neoplastic digestive diseases. Important elements complicating a decompensated liver cirrhosis are ascites, hepatic encephalopathy, digestive bleeding and jaundice. Acute liver failure (ALF) is the expression of a clinical state, that is common to many conditions sharing severe liver structural and functional impairments. In patients affected by decompensated liver cirrhosis, ALF could be triggered by several factors, while the death is caused by bleeding episodes, hepato-renal syndrome, spontaneous bacterial peritonitis or hepatocarcinoma. In patients affected by chronic liver diseases, the diagnosis of ALF is based on progressively increasing jaundice, encephalopathy and coagulopathy. Recent clinical trials have evaluated the efficacy of extrahepatic liver support systems, either artificial or bio-artificial, in treating episodes of ALF in chronic liver patients. The preliminary results indicate a potential use of such systems in blood detoxification, but they also showed limits in increasing patient survival. PMID- 11014010 TI - [Hybrid (bioartificial) systems for liver support. History and current developments]. AB - Attempts to develop liver support systems for the treatment of acute liver failure patients have ranged in the past, from the use of hemodialysis, or plasma exchange, or activated charcoal particles and synthetic resins, as well as the use of bioreactors loaded with liver tissue. However, no system demonstrated a significant improvement of patient survival, nor has achieved a wide clinical use. Liver transplantation remains the only treatment for severe hepatic failure that can improve patient survival. On the other hands, the chronic scarcity of organs for transplantation, leads to an urgent necessity of liver support systems. In this paper, we reviewed the historical experience and current status of artificial liver support systems, with particular emphasis on the so-called hybrid or bioartificial liver, in which to the traditional artificial components, such as selective membranes, charcoal particles and resins, isolated hepatocytes are used. PMID- 11014011 TI - [Artificial liver support systems: state of the art]. AB - Previous attempts in improving the outcome of acute liver failure patients were based on extracorporeal artificial systems such as hemodialysis, hemoperfusion, plasmaperfusion, hemoadsorbtion and plasma exchange. Despite more than 30 years of research and development in this field, an artificial liver has still not become a reality, since purely artificial support systems have shown only minor improvement in patient's survival, and orthotopic liver transplantation is the only effective cure. This review takes a critical look at past and present concepts for an artificial liver support system, evaluating the advantages and disadvantages of the aforementioned techniques. Progress in methods and techniques for maintaining long-term hepatocyte culture in vitro indicate the potential use of isolated liver cells in bioartificial liver support systems. Recent studies suggest the use of traditional artificial liver support systems, such as hemoadsorbtion or plasma-absorbtion on activated charcoal particles or synthetic ion-exchange or neutral resins to improve and prolong the efficiency of the so-called hybrid or bioartificial liver, in which isolated hepatocytes are used. PMID- 11014012 TI - [Isolated hepatocytes and their potential therapeutic use]. AB - Isolated hepatocytes in culture represent an idoneous system for the study of liver physiology and metabolism. Furthermore, they are also widely utilized in pharmacological and toxicological study, in evaluating xenobiotic substance effects on the liver. In this paper, we reviewed the enzymatic methods for liver cell isolation in some mammalian species, as well as the techniques for qualitative and quantitative evaluation of cell number, vitality, purity, morphology and function. Recently, there has been a renewed interest in hepatocyte transplantation and hepatocyte-based liver support systems. From a clinical point of view, isolated hepatocytes could be useful in temporarily substituting an acutely damaged liver, a liver affected by a chronic pathology, or to correct an inherited liver disease carrying a severe metabolic derangement. Early experimental results of allogeneic hepatocyte transplantation, as well as the first clinical trials of bioartificial liver support systems employing xenogeneic hepatocytes are promising and contribute to maintain that interest in liver cell isolation and purification methods. PMID- 11014013 TI - [In vitro experimentation of a new model of radial flow bioreactor containing isolated hepatocytes]. AB - Hepatocyte based artificial liver support systems are under investigation to support acute liver failure patients. The main purpose of such systems is to serve as a bridge to liver transplantation, or to promote spontaneous liver recovery. Limitation in mass-transfer capacity makes hollow-fiber bioreactors unsuited for long-term functioning of hybrid devices. We developed a novel radial flow bioreactor in which the fluid perfuses the module from the center to the periphery, after having diffused through a space occupied by a three-dimensional structure filled with the hepatocytes. Five grams of freshly isolated porcine hepatocytes were seeded into uncoated, woven-non woven, hydrophilic polyester fabric, overlaid by two polyethersulfone membranes. Liver cells were perfused with 37 degrees C-warm, oxygenated, serum-free tissue culture medium, in which NH4Cl and Lidocaine were added at the final concentration of 1 mM and 60 micrograms/ml, respectively. Ammonium chloride removal, urea synthesis, monoethylglycinexylide (MEGX), pO2, pCO2, and pH were measured throughout the 14 day duration of the study. In a separate set of experiments, a scaled-up version of the radial flow bioreactor containing 150 grams of cells was perfused for 7 h with recirculating human plasma and MEGX production was monitored. During the 2 weeks of the study, an increasing production of urea was paralleled by constant ammonium removal. MEGX concentration after Lidocaine addition increased throughout the 14 days of perfusion with tissue culture medium, as well as after 7 hour perfusion with human plasma. Under transmission and scanning electron microscopy cells appeared attached to the polyester and one to each other, displaying ultrastructural features typical of functioning hepatocytes. Our study showed that liver cells were metabolically active when perfused into the radial flow bioreactor. This configuration allowed close contact between media, or plasma, and cells at a physiological flow rate, by equalizing the concentration of the perfusing components, including O2, throughout the module. Our results suggest a potential use of this system for temporary extracorporeal liver support in acute hepatic failure patients. PMID- 11014014 TI - [Therapeutic approach in Riedel thyroiditis]. AB - Riedel's invasive fibrous thyroiditis is a rare disorder of unknown origin with progressive extension and invasion of adjacent structures. Clinically it is impossible to distinguish between Riedel's/thyroiditis and other diseases as undifferentiated carcinoma, Hashimoto's disease etc... The patients often have dyspnea, dysphagia, paralysis of the vocal cord; fine needle puncture-aspiration and biopsy themselves may be insufficient, Surgical treatment depends on the stage of the disease, when both lobes are involved generous wedge resection of the isthmus may be the treatment of choice to relieve tracheal compression; in earlier stages radical operation are considered. Corticosteroid treatment in Riedel's thyroiditis, as multifocal disease has been successfully used. Other drugs with antifibrosing actions have also utilised in small groups of patients with encouraging results. PMID- 11014015 TI - [Our experience with duodenal tumors]. AB - The authors report 3 cases of benign duodenal tumors and one case of duodenal cancer. In two cases the benign tumors were adenomatous polyps, in one case with signs of dysplasia. One case of benign tumor was a lipoma. All the three benign tumors were removed through a duodenotomy, in one case associated with a sphincteroplasty. The case of duodenal cancer, arisen in a patient operated since 24 years with a gastric resection, was treated with a Whipple technique but for the very old age and the bad health conditions, the patient died in 11th post operative day for myocardium infarct. PMID- 11014016 TI - [Indications for the surgical treatment of cavernous angioma of the liver in adults. Report of a case]. AB - The authors report a case of giant cavernous hemangioma of the liver, almost entirely extrahepatic, bulging from the inferior surface of the right lobe into the abdomen, treated surgically on account of worsening symptoms referable to continuing growth. A thorough search of the literature allows a critical review of the surgical indication in this benign pathology, rarely complicated by significant clinical events. Surgery is indicated, in lack of relevant complications, only when significant symptoms, continuing or worsening, are undoubtedly referable to the presence of the angioma. PMID- 11014017 TI - [Retrospective analysis of the use of prophylactic drainage of the pelvis after anterior resection of the rectum]. AB - Although the use of drains is common in clinical practice, its real role in the prophylaxis and therapy of postoperative complications is still not clear. In the literature we can find both supporters of drains, and many opponents who consider their use unnecessary and sometimes even dangerous. In fact, during new experimental and clinical studies, it was impossible to determine the usefulness of prophylactic abdominal drain, at least in the case of colo-rectal anastomosis, it has been demonstrated that use of drains limits the risks of an anastomotic leakage but, in some cases, the same drains could be the cause of some of the complications that should be avoided. Although there is a considerable theoretical and practical evidences in favour of drainage, the dispute about "to drain or not to drain" the peritoneal cavity after elective colo-rectal surgery remains open. This retrospective study made on 150 patients operated on elective surgery for rectal cancer demonstrates that prophylactic drain does not significantly influence the general rate of leakage (3.15% for group A and 5.45% for group B, p > 0.1); in two of the three fistulas in patients with drains, the drains have permitted the diagnosis, but have not permitted the reduction of the number of operations for fistulas. PMID- 11014018 TI - [Unusual case of benign neoplasm of the breast mimicking a carcinoma: granular cell tumor. Case report]. AB - Granular cell tumors are rare, usually benign neoplasms of soft tissues which most commonly occur in the tongue, skin and subcutaneous tissue. Although the histogenesis is still object of debate, recent immunohistochemical studies and ultrastructural findings support the origin of this neoplasm from the peripheral nervous tissue, most likely from Schwann's cells. Occurrence of this neoplasm in the breast, although uncommon, warrants special attention, since its clinical, mammographic and pathological appearances on frozen sections "may often closely resemble" hose of breast malignancy. The authors analyze and commenton, with special reference to clinical aspects and surgical treatment, a case of benign granular cell tumor of the breast occurring in a 42 year-old woman. The mammographic and clinical findings suggested a breast carcinoma. The correct diagnosis was established by definitive microscopic examination of the paraffin embedded specimens and the treatment was a simple local excision of the tumour and a small rim of surrounding breast parenchyma. Although the granular cell tumor of the breast is a rare entity, surgeons and pathologists should be aware of its existence in order to avoid inappropriate radical surgery not justified by the benign behavior of the neoplasm. PMID- 11014019 TI - Spontaneous rupture of a giant hemangioma of the liver. AB - Hemangiomas are frequent benign tumors of the liver. Symptoms (abdominal pain and fullness) are mostly seen in giant lesions. Rupture is the most severe complication, can occur spontaneously, with intraperitoneal bleeding, in 1-4% of hemangiomas and has been described in about 30 cases in the international literature with a high mortality (about 60%). This complication is the principal indication for surgery. Although spiral CAT scan and MR are actually the most efficacious imaging methods for study of liver hemangiomas, after Echography, emergency techniques that allows a simultaneous therapeutic approach--as is angiography--are preferable. Trans-arterial embolization (TAE) is in fact useful to stop bleeding and then to perform a safer surgery. A successful embolization can delay the surgical resection of the hemangioma for the time necessary to recover from the hemodynamic distress. Aside from the success of angiographic approach, surgery remains mandatory, effective in stopping the bleeding and in preventing re-bleeding or other complications of TAE such as abscess, fever, etc.. Intraoperative echography currently is the best method to identify vasculo- biliary anatomy and to perform a correct resection. The absence of risk factors for spontaneous rupture of liver hemangiomas, makes this event unpredictable. The best treatment for non-ruptured hemangiomas is still controversial but surgery is usually limited to symptomatic tumors larger than 10 cm. PMID- 11014020 TI - [Adenocarcinoma arising on colo-rectal endometriosis: clinico-pathological considerations]. AB - Colo-rectal endometriosis represents a rare pathological event. Exceptionally, a malignant colo-rectal endometrioid carcinoma arising from endometriosis has been reported. In the present case, we describe a 32 year-old woman with clinical and radiological signs of large bowel sub-occlusive syndrome. Histologically, a sigmoid endometrioid adenocarcinoma associated with submucosal foci of eutopic endometrium was observed. Moreover, endometriotic tissue was observed at the level of the colonic left flexura and appendix. Colo-rectal endometriosis must be considered among risk factors for the development of large bowel adenocarcinoma in young female patients. PMID- 11014021 TI - [Umbilical endometriosis: report of a case and review of the literature]. AB - The authors present a clinical of umbilical endometriosis in a young woman. This patient never underwent any operation or diagnostic abdominal procedures. The case is very interesting because of problems concerning differential diagnosis with other umbilical and periumbilical diseases and pathogenetic theories. In this study the authors suggest a hypothesis to explain the localization of ectopic endometrium in this site. We review the literature, with some comments on therapeutic options; finally we compare the medical and the surgical approach and we suggest an advisable therapy. PMID- 11014022 TI - [Ileal occlusion caused by enterolith migrated from Meckel's diverticulum]. AB - The authors present a case report of an intestinal obstruction due to a relatively big coprolite migrated from a large Meckel's diverticulum to the distal Ileum. The patients apparently healthy and a vegetarian, complained on admission of the absence of emission of faeces and gas since four days before with noticeable abdominal distension. In the physical examination he presented intestinal meteorism, a hard abdomen, painful on deep palpation in the median quadrants, especially in the epigastric and mesogastric ones. The abdominal X-RAY in the standing position confirms: an occlusive state with numerous liquid levels in the Ileum. Because of a worsening of the symptomatology and the appearance of generalized comprimission, two days later an exploratory laparotomy was performed. The intervention showed the presence of a Meckel's diverticulum with approximately 10 cm in length, with an ample neck, the distal Ileum for approximately 15 cm in dilation returned rapidly to a normal calibre, after a pastous endoluminal formation borne in the Meckel's diverticulum (Meckel resection presented actually a large niche at the fundus level with a eroded wall) and migrated in the distal Ileum where it could cause the obstruction. In the present case it is probably useful to perform a preoperative CT scan in order to get a precise etiology and perform an ascending Colonscopy, so avoiding a surgical procedure. According to the authors a CT scan is indicated in all cases of intestinal occlusion of unknown case, in order to have a more precise definition of the physiopathology of the occlusion. PMID- 11014023 TI - Ready, steady, go for HCV antigen testing? PMID- 11014024 TI - Implications and importance of genotyping cryptosporidium. PMID- 11014025 TI - Serological diagnosis of syphilis. PHLS Syphilis Serology Working Group. AB - The availability of an increasing number of enzyme immunoassays (EIAs) for detecting syphilis antibodies makes it appropriate to review approaches to syphilis serology and to assess the role of syphilis EIAs in routine diagnostic microbiology laboratories. This paper summarises the principles and practice of syphilis serology and provides recommendations on the use of laboratory tests for syphilis in UK diagnostic microbiology laboratories. The main recommendations are summarised in a testing algorithm. Treponemal EIAs are an appropriate alternative to the use of combined Venereal Disease Research Laboratories/rapid plasma reagin and Treponema pallidum haemagglutination assay (TPHA) tests for screening for syphilis. If a treponemal EIA is used for screening an alternative treponemal test, such as TPHA, should be used for confirmatory testing. The fluorescent treponemal antibody-absorbed test is probably best reserved for specimens giving discrepant results. Such specimens may be referred to the PHLS laboratories that provide confirmatory treponemal testing for reference testing and to facilitate collection of surveillance data on what remains an important public health problem. PMID- 11014026 TI - Guidelines for the microbiological quality of some ready-to-eat foods sampled at the point of sale. PHLS Advisory Committee for Food and Dairy Products. AB - These guidelines for the microbiological quality of ready-to-eat foods represent a revision and expansion of guidelines first published by the PHLS in September 1992 and revised in March 1996. The latest guidelines incorporate many of the constructive comments received from food examiners and other microbiologists within and outside the PHLS and from environmental health officers throughout the United Kingdom. This document reviews the changes and the reasons they were made and sets out the new guidelines. It also clarifies the role of food examiners in interpreting the microbiological results of formal samples. PMID- 11014027 TI - Outbreak of meningococcal disease in Rotherham illustrates the value of coordination, communication, and collaboration in management. AB - On New Year's Eve 1998, two teenagers from the same school in Rotherham died of serogroup C meningococcal septicaemia. A third linked case occurred three days later. Over eight days starting on New Year's Eve, a further five cases of meningococcal disease arose across Rotherham district, four of whom were severely ill with serogroup C meningococcal septicaemia. Intense media interest, high levels of public concern, and anxieties caused by several 'false alarm' cases added to the challenge faced by a small health district. This article describes the epidemiology of the incident and policy decisions made, but focuses on the operational aspects of outbreak management (coordination, communication, and collaboration), an area often neglected in outbreak reports. PMID- 11014028 TI - Carriage of Neisseria meningitidis in residents and staff at a residential home for elderly people following a case of invasive disease. AB - A study of the carriage of Neisseria meningitidis among staff and residents of a 'closed community' (a residential home for elderly people) was conducted after a resident developed invasive meningococcal disease. All 39 other residents and 49 staff who worked at the home during the previous seven days were offered a throat swab and 38 residents and 49 staff consented. Two residents (none in the index case's social group) and one staff member were found to be carrying N. meningitidis, all phenotypically distinct from the infecting isolate. Thus, all four individuals carried different organisms and the index case was sporadic. Our findings suggest that residents or staff in long stay residential settings where a sporadic case occurs need not be offered meningococcal prophylaxis unless they fulfil the existing definition of 'close personal contacts' of the case. PMID- 11014029 TI - Optimising investigation of meningococcal disease to enhance surveillance. AB - Efforts were made to implement guidelines for microbiologists in order to optimise investigation and improve laboratory confirmation in children admitted to hospital with suspected meningococcal disease. In 1998, 75% of diagnoses (30/40) were confirmed microbiologically, compared with 51% (209/411) of cases elsewhere in the West Midlands. Thirty-two of the children admitted to our paediatric unit (80%) were investigated according to the guidelines; 94% of these cases were laboratory confirmed. Microbiological confirmation of clinical meningococcal disease is all the more important with the advent of mass vaccination. Our study shows that implementation of guidelines for the investigation of clinically suspected meningococcal disease improves the rate of confirmation. PMID- 11014030 TI - Outbreak of group C meningococcal infection affecting two preschool nurseries. AB - Five cases of meningococcal infection caused by Neisseria meningitidis group C type 2a, subtype P1.5, P1.2 occurred within six days. Two of the affected children attended the same preschool nursery. The mother of two other children at the nursery was affected, as were the mother and sibling of another child (not a case) at the nursery. The 'sibling' case attended a different nursery. As the situation evolved, the control aspects of the outbreak changed. Antibiotic prophylaxis and vaccination were given to all staff and children attending both nurseries, and to parents, siblings, and household contacts of all children attending the first nursery. PMID- 11014031 TI - Outbreak of tuberculosis associated with a church. AB - Investigation of an outbreak of tuberculosis (TB) in a West Midlands health district in 1999 revealed spread in an extended family network and to church contacts. Within the family four cases of smear positive TB, four cases of smear negative infection, and 14 cases requiring chemoprophylaxis were identified. One of the infectious cases visited a local church on two occasions, which resulted in a further 16 cases of infection including one case of tuberculous meningitis. DNA fingerprinting of isolates from five culture positive cases indicated that the same strain of Mycobacterium tuberculosis was responsible. This outbreak is a reminder that while outbreaks of TB usually arise within households or family networks, where close contact over extended periods provides more opportunity for exposure, community outbreaks of TB can occur after only causal contact. PMID- 11014032 TI - Limitations of national guidelines in the management of an outbreak of tuberculosis. AB - National guidelines offer no clear definition of a close (non-household) contact of tuberculosis. The resulting lack of distinction between close and casual contacts may lead to excessive screening in outbreaks of pulmonary tuberculosis in the United Kingdom. Poor compliance with chemoprophylaxis suggests that priority should be given to the follow up of high risk contacts, who should be more clearly defined. A review of the management of a family outbreak of tuberculosis illustrates the problem. Fifty-nine out of 213 non-household contacts screened for infection had grade 3 and 4 Heaf reactions and none developed clinical tuberculosis within one year. Three of the 59 did not attend for chest radiography and a further 11 patients did not attend the outpatients appointment offered. Twenty-three took chemoprophylaxis for three months, 14 did not complete chemoprophylaxis, six (11%) declined it and two were offered x-ray follow up. PMID- 11014033 TI - Survival after diagnosis of AIDS among adults resident in the United Kingdom in the era of multiple therapies. AB - The epidemiology of HIV and AIDS in the United Kingdom (UK) has changed markedly since highly active antiretroviral therapy (HAART) was introduced in 1996. HAART including protease inhibitors has considerably improved survival from AIDS diagnosis. The number of deaths of individuals with HIV infection in the UK, reported within 12 months of the end of the year of death, have decreased between 1995 and 1998. Concurrently AIDS diagnoses, reported within 12 months of the end of the year of diagnosis, have declined whilst diagnoses of HIV infection, similarly reported, have risen. Data from 13,689 adult AIDS cases diagnosed up to the end of 1996 were analysed. The overall median survival from AIDS diagnosis to death was 19.3 months. Over 50% of the cases diagnosed in 1996 were alive at the end of the survey therefore median survival exceeds 24 months, the maximum follow up time for the cohort. The opportunity for receiving HAART was modelled in three time periods: pre-multiple therapies (before September 1995), multiple reverse transcriptase inhibitor therapy available (September 1995 to March 1996), and multiple therapy including protease inhibitors available (April 1996 onwards). Survival rates improved significantly among female heterosexuals and men who have sex with men when multiple therapy including protease inhibitors became available. PMID- 11014034 TI - Evaluation of health information provided on the Internet by airlines with destinations in tropical and sub-tropical countries. AB - The travel industry is being called upon increasingly to provide health information for its clients. Websites of all 73 international airlines with tropical or sub-tropical destinations accessible via the International Air Transport Association homepage were evaluated for the quality of their health advice. Seven of the 73 homepages contained health information, six of which addressed health problems that might be encountered when travelling to tropical countries. Fifty-five of the 73 airlines had functional email addresses and were asked for pre-travel health advice on malaria prophylaxis using a fictitious scenario. Among the 25 airlines that replied, only half referred to malaria and a third gave wrong information about malaria endemicity in the destination area. The only two airlines that recommended a specific drug regimen correctly reflected the medical situation described in our fictitious email. In general, international airlines are not set up to provide health information. PMID- 11014035 TI - Acceptability of health status questionnaire screening and vaccination among Kosovar refugees. AB - During 1999, following the war in Kosovo, thousands of Kosovar refugees were received by different countries. The public health reception protocols for these refugees varied widely between and within reception countries. We surveyed a group of Kosovar refugees received in Bolton to assess the acceptability of vaccination and health status questionnaire screening. We also examined the refugees' opinions about the confidentiality of their medical history and their willingness to take part in research. Fifteen out of 29 people responded. Most respondents were happy with the interventions. PMID- 11014036 TI - Investigation and management of sporadic gastrointestinal infections with potentially Vero cytotoxin producing Escherichia coli in Scotland. AB - Recognition of the potential of Escherichia coli O157 and other Vero cytotoxin producing E. coli (VTEC) organisms to cause serious disease led to the recommendation that all diarrhoeal stool specimens be examined for E. coli O157. National guidelines exist for the testing and exclusion of cases and contacts of VTEC infection. A survey was conducted to discover the extent to which these recommendations are followed in Scotland by asking about current practices for public health management of identified cases and laboratory investigation of E. coli infection. About two thirds of Scottish health boards followed national guidelines for testing and exclusion of cases and contacts of VTEC O157 infection. Most laboratories tested all diarrhoeal stools for E. coli O157 but detection methods varied and a minority tested selected stools for non-O157 E. coli serogroups. Standardisation of policies for laboratory testing of VTEC infection would improve national surveillance. Adherence to evidence based guidelines would standardise public health management of VTEC infections in Scotland. PMID- 11014037 TI - Factors associated with seeking emergency treatment following suspected chemical contamination of a leisure pool. AB - A leisure pool was evacuated when children complained of breathing difficulties, cough, and eye irritation subsequently thought to be due to high chloramine levels. The duration of the suspected contamination before the evacuation was unknown: it was suggested that some subsequent attendances at accident and emergency (A&E) departments had been prompted by news reports of the incident. The extent and nature of symptoms, the cause of the incident, and the impact of media reporting were investigated with the help of a postal questionnaire. One hundred and thirty-seven people (all but six under 20 years of age) attended A&E departments after the incident, most commonly with sore eyes (79%), cough (76%), and sore throat (71%). The number of different symptoms was associated with the length of time spent at the pool, but not with being present at the time of the evacuation or with having heard about the incident on radio or television. PMID- 11014038 TI - Outbreak of giardiasis in a daycare nursery. AB - Seven children and one member of staff in a nursery in Kent became ill with Giardia lamblia infection in three months in a nursery in Kent and three further children were found to carry giardia. Person to person spread among young children who were not toilet trained appeared to be responsible. All staff and children were screened for infection, and the outbreak was controlled by reinforcing hygiene measures, and treating microbiologically confirmed cases and carriers with metronidazole. PMID- 11014039 TI - Chickenpox increasingly affects preschool children. AB - Between 1983 and 1998, age specific incidences of chickenpox derived from consultations with general practitioners taking part in the Royal College of General Practitioners Weekly Returns Service doubled in children aged 0 to 4 years, halved in children aged 5 to 14 years, and fell by almost a third in adults aged 15 to 44 years. This downward shift in age of contracting chickenpox may be a result of increased social contact between preschool children. PMID- 11014040 TI - Hepatitis B immunisation of infants at risk. AB - A retrospective cohort review survey assessed hepatitis B immunisation uptake in at-risk infants over 4 years and a GP questionnaire identified the problems of poor communication and uncertainty of responsibility for immunisation as barriers to successful implementation of the immunisation programme. Incorporating appointments for hepatitis B immunisation within the national child immunisation recall system would help improve uptake but, in the meantime, a manual appointment system could be incorporated within existing child immunisation systems. PMID- 11014041 TI - Needlestick injuries and hepatitis B virus vaccination in health care workers. AB - An analysis of 2646 needlestick injuries in hospitals in the Greater Manchester area between April 1992 and April 1999 was carried out. Ten per cent of members of staff injured in these incidents had never been vaccinated against hepatitis B virus (HBV) and 27% of those who had been vaccinated had no anti-HBs (< 10 IU/L). Although few health care workers were at risk of transmission of HBV through needlestick incidents in this study (0.6% (12/2084) of all source patients were HBsAg positive; 9 HBeAg positive, 7 anti-HBe positive), the large number of members of staff who were not protected from HBV infection indicates a need for occupational health departments to reinforce HBV vaccination policies. PMID- 11014042 TI - Pertussis vaccine efficacy in Birmingham. PMID- 11014043 TI - Screening for chlamydia. PMID- 11014044 TI - Procuring more organs for transplantation. PMID- 11014045 TI - Bioinformatics from the perspective of the public health laboratory. PMID- 11014046 TI - Production of standard operating procedures, algorithms, and guidance notes for virology and serology. PMID- 11014047 TI - Manual handling at work. PMID- 11014048 TI - Outbreak of cryptosporidiosis in Northern Ireland. PMID- 11014049 TI - National increase in Salmonella typhimurium DT104--update. PMID- 11014050 TI - People focus on optimistic scenarios and disregard pessimistic scenarios while predicting task completion times. AB - Task completion plans normally resemble best-case scenarios and yield overly optimistic predictions of completion times. The authors induced participants to generate more pessimistic scenarios and examined completion predictions. Participants described a pessimistic scenario of task completion either alone or with an optimistic scenario. Pessimistic scenarios did not affect predictions or accuracy and were consistently rated less plausible than optimistic scenarios (Experiments 1-3). Experiment 4 independently manipulated scenario plausibility and optimism. Plausibility moderated the impact of optimistic, but not pessimistic, scenarios. Experiment 5 supported a motivational explanation of the tendency to disregard pessimistic scenarios regardless of their plausibility. People took pessimistic scenarios into account when predicting someone else's completion times. The authors conclude that pessimistic-scenario generation may not be an effective debiasing technique for personal predictions. PMID- 11014051 TI - How serious are expressions of protected values? AB - People think that some things that they value should be protected from trade-offs with other things. For example, people think that no economic gain is great enough to justify clear-cutting old-growth forest. The authors probed the stability of these protected values (PVs) in several ways. Subjects were asked to think of counterexamples, and this had some effect on PVs. Subjects were then asked how they would resolve conflicts between 2 PVs. Resolutions did not differ from those between other values, but subjects tended to feel that conflicts between PVs did not occur in reality. Despite people's claims that PVs are unchanged by variation in quantity, expression of PVs was reduced when the magnitude or probability of the violation of a PV was smaller. In summary, PVs appear to be labile and amenable to challenge. Despite earlier concerns, apparent PVs may not always preclude the use of valuation measures in cost-effectiveness analysis or negotiated agreement on controversial issues. PMID- 11014052 TI - Asking questions can change choice behavior: does it do so automatically or effortfully? AB - The present research uses a technique that permits unique estimation of both automatic and effortful processes in the question-behavior link. Results show that individuals asked to report behavioral intent (vs. those not asked) are more likely to choose options that are highly accessible and positively valenced, regardless of cognitive resources available at the time of processing. This suggests that the effect of intent questions on subsequent behavior is primarily the result of automatic, as opposed to effortful, processing. Practically, this suggests that efforts to debias this robust effect need to affect nonconscious processes and adjust for the automatic impact of being asked an intention question on respondents' behavior. PMID- 11014053 TI - Process versus content in eyewitness metamemory monitoring. AB - Three studies (Ns = 200, 135, and 187 college undergraduates) contrasted process versus content accounts of eyewitness metamemory monitoring. Subjective vividness, a cue related to memory content, was a better predictor of confidence and accuracy than were cues related to the retrieval process. Participants who were asked to recall, rather than recognize, event details displayed greater insight into accuracy, primarily because vividness was a more valid accuracy cue under recall conditions. Results reinforce the value of recall-based protocols for eliciting eyewitness testimony and suggest some specific conditions (e.g., yes-no recognition) under which investigators should be especially cautious in relying on confidence to infer accuracy. In addition, results point to a general framework for understanding moderating effects on eyewitness metamemory accuracy. PMID- 11014054 TI - When to inspect? Recurrent inspection decisions in a simulated risky environment. AB - Participants scheduled inspections to detect costly events for which they were repeatedly at constant risk (probability of event onset) within a computerized environment. They were responsive to risk variations, conveyed either in advance or by experience with inspection outcomes, although experiencing unannounced increases in risk affected inspections more than experiencing unannounced decreases. Participants responded to variations in cost (time varying or fixed) when the effects were made perceptually salient. Compared with a normative model (R. L. Klatzky, D. M. Messick, & J. Loftus, 1992), some conditions showed near optimal inspecting or had flat payoff functions that tolerated observed departures from optimal performance. Costly departures occurred particularly when combined cost and risk levels caused optimal responses to be extreme (always or never inspect). Results assess people's processing of relevant variables and indicate circumstances in which they may set substantially nonoptimal inspection schedules. PMID- 11014055 TI - The impact of sleep deprivation on decision making: a review. AB - Few sleep deprivation (SD) studies involve realism or high-level decision making, factors relevant to managers, military commanders, and so forth, who are undergoing prolonged work during crises. Instead, research has favored simple tasks sensitive to SD mostly because of their dull monotony. In contrast, complex rule-based, convergent, and logical tasks are unaffected by short-term SD, seemingly because of heightened participant interest and compensatory effort. However, recent findings show that despite this effort, SD still impairs decision making involving the unexpected, innovation, revising plans, competing distraction, and effective communication. Decision-making models developed outside SD provide useful perspectives on these latter effects, as does a neuropsychological explanation of sleep function. SD presents particular difficulties for sleep-deprived decision makers who require these latter skills during emergency situations. PMID- 11014056 TI - What's new in the treatment of cerebrovascular disease?: an endovascular perspective. AB - Surgical therapy currently allows for the correction of many pathologic conditions affecting the vasculature of the central nervous system. Recent advances in technology have given the cerebrovascular specialist further options in the minimally invasive sphere. Endovascular techniques, complimentary to surgery in some conditions, are poised to replace conventional open surgery in others. A review of current interventional radiological procedures for the treatment and prevention of ischemic and hemorrhagic stroke are herewith presented. PMID- 11014057 TI - Physician-based tobacco dependence interventions: review and clinical practice recommendations. PMID- 11014058 TI - Ten commandments of risk management. PMID- 11014059 TI - MSMA joins Medem to provide members and their patients with authoritative and secure online information and communications. PMID- 11014060 TI - Putative peripheral neuropathy in suckling piglets. AB - Over a period of almost 2 years, a progressive motor disturbance was found to occur in 20-50% of the litters of both primiparous and multiparous sows in a large pig herd of 1000 sows. The motor disturbance sometimes affected the entire litter; however, in most cases only a few piglets per litter were affected. The clinical signs appeared at 3-5 days of age and consisted of difficult movement followed by anteflexion or retroflexion of the tarsal joints or 'rabbit-like posture'. Subsequently, primarily after weaning, inflammatory and necrotic lesions developed on the paralysed limbs as a result of secondary infections of injuries. The tibial nerve and the common fibular nerve of recently affected (5- to 6-day-old) piglets showed degeneration, demyelination and necrosis of some of the nerve fibres, accompanied by restorative changes in more chronic cases. The central nervous system, bones, skeletal muscles, tendons and joints showed no lesions that could have accounted for the symptoms of motor disturbance. Aetiological investigations excluded the possibility of lead, copper and cadmium toxicity. Vitamin B2 administered orally at 1 day old proved to be ineffective. The disease did not develop in piglets of sows kept at another farm under the same management and fed a diet prepared according to an identical formula from the same ingredients as those used on the affected farm, but with no milk whey added. This raised the suspicion of triaryl phosphate (TAP) poisoning, but this was found not to be the cause of the disease. New boars had not been brought to the farm in the year preceding the onset of disease, and the disease could not be linked to a specific boar or boar line. The aetiology of the disease has remained unclear. PMID- 11014061 TI - Characterization of lesions caused by a South American virulent isolate ('Quillota') of the hog cholera virus. AB - In this study, macroscopic and histopathological lesions produced by a virulent South American isolate ('Quillota') of hog cholera virus were studied. The virus was inoculated in doses of 10(5)TCID50 in each of 35 pigs of 20 kg live weight. The animals were slaughtered from 4 to 18 days post-inoculation. The presence of virus antigens in lymphatic tissue was confirmed by both direct immunofluorescence and Avidin-Biotin-Peroxidase techniques in formalin-embedded tissue samples. Histological sections were stained with haematoxylin-eosin and Mallory's phosphotungstic acid haematoxylin methods. The 'Quillota' isolate used in this study caused a disease characterized by vascular lesions (splenic infarcts, haemorrhages in the lymph nodes and the urinary system and disseminated microthrombosis), and necrosis of lymphocytes, particularly in the B-areas of the lymphoid organs, lesions that are characteristic of the acute form of the disease. Other lesions observed were a non-purulent meningoencephalitis, the necrosis of the epithelial cells of tonsils, the presence of fibrin nets in the red pulp and a marked thickening of the alveolar septa. PMID- 11014062 TI - The modification of fluorescent antibody virus neutralization (FAVN) test for the detection of antibodies to rabies virus. AB - The fluorescent antibody virus neutralization test (FAVN) for the detection of antibodies against rabies virus was modified by using a monoclonal anti-rabies antibodies and peroxidase anti-mouse conjugate instead of a fluorescent anti rabies conjugate. The results were read on an automatic multi-channel spectrophotometer. A total of 182 serum samples from dogs were tested by both the original and modified FAVN methods and the results were compared. Good correlation was found between the two tests. Practically, the modified FAVN test was quicker and could be used for a larger number of samples. PMID- 11014063 TI - Immunoglobulin G class identification from wild ungulates by cross-reactivity with antisera to domestic animals. AB - Seven species of Spanish ungulates were tested for the presence of homologous immunoglobulin G (IgG) with a gel-diffusion test using bovine, ovine, caprine and porcine IgG antisera. Homologous ovine and caprine IgG were detected in sera from chamois (Rupicapra pyrenaica), Spanish ibex (Capra pyrenaica hispanica), mouflon (Ovis orientalis musimon), red deer (Cervus elaphus), fallow deer (Dama dama) and roe deer (Capreolus capreolus). Homologous porcine IgG was detected in wild boar (Sus scrofa) serum. Immunoelectrophoretic assays were performed to compare the electrophoretic mobility of IgG from domestic and wild species. PMID- 11014064 TI - Isolation of Candida rugosa from turkeys. AB - The present study describes the isolation of Candida rugosa from young turkeys that died 10 days after the end of a therapeutic treatment for a recent outbreak of coccidiosis. Candida rugosa was isolated from the digestive tract of all the birds examined. This isolation is the first in turkeys and corroborates the fact that C. rugosa is an opportunistic yeast which circumvents host defences when other predisposing factors are present. PMID- 11014065 TI - The sensitivities of various erythrocytes in a haemagglutination assay for the detection of psittacine beak and feather disease virus. AB - The erythrocytes of various species were tested in psittacine beak and feather disease (PBFD) virus haemagglutination (HA) and haemagglutination inhibition assays to determine which are suitable for use in these assays. HA activity was observed for erythrocytes of the salmon-crested cockatoo, the sulphur-crested cockatoo, the umbrella cockatoo, the goffin's cockatoo and the cockatiel, with differences amongst individuals within species, but not for erythrocytes of humans, the pig, the guinea pig, the chicken, the goose, the rose-ringed parakeet or the budgerigar. Anti-PBFD virus rabbit sera inhibited the virus-induced agglutination of erythrocytes, confirming the specificity of HA activity. This suggests that selection of suitable psittacine species as well as suitable individuals within a species is necessary when obtaining erythrocytes for the PBFD virus HA assay. PMID- 11014066 TI - Effectiveness of doxycycline in the prevention of an experimental infection with Actinobacillus pleuropneumoniae in pigs. AB - The effectiveness of medication with doxycycline in feed in the control of pleuropneumonia in pigs was tested using an Actinobacillus pleuropneumoniae serotype 1 aerosol challenge model. Two groups of 10 animals were used for the challenge, a 'medicated group' and an 'unmedicated group'. A third group of four animals was used as a 'control group'. Pigs from the medicated group were provided with feed containing 250 p.p.m. doxycycline (HIPRAMIX/DOXI) for 8 consecutive days and were challenged on the fifth day of treatment. No clinical signs were observed in pigs from the 'control group'. Four animals from the 'unmedicated group' died within the first 48 h after challenge with clinical and lesional evidence of an acute form of pleuropneumonia. Clinical signs of animals surviving the first 48 h were progressively less severe and showed lesions similar to those described for subacute-chronic forms of the disease. However, only one animal from the 'medicated group' showed clinical signs of a chronic form of pleuropneumonia. Reisolation of A. pleuropneumoniae was more evident from lung tissues of animals fed the doxycycline-free feed (70%), coinciding with the presence of both acute and subacute lesions. However, the micro-organism could be reisolated from only one animal which belonged to the 'medicated group'. It is concluded that the treatment of pigs with 250 p.p.m. doxycycline (HIPRAMIX/DOXI) prevents disease caused by A. pleuropneumoniae. PMID- 11014067 TI - Influence of antibiotics used as feed additives on the immune effect of erysipelas live vaccine in swine. AB - To investigate the influence of antibiotics used as feed additives on the immune response to erysipelas live vaccine, the pig inoculation test was applied. Avilamycin, oxytetracycline quaternary salt, enramycin, virginiamycin and tylosin phosphate were selected as test antibiotics. Five experimental feeds containing each antibiotic at the highest concentration permitted for feed additives in Japan, and the basal diet lacking antibiotics were examined. Twenty-nine pigs were divided into six groups. At first all the groups were fed with the antibiotic-free basal diet for 7 days, and then each group received the experimental feeds. On the 14th day after feeding with test feeds all the pigs, except for one control pig in each group, were immunized with the vaccine and all the pigs were then challenge-exposed to a virulent strain of Erysipelothrix rhusiopathiae 14 days after vaccination. The clinical response was observed every day for 14 days. In all the groups, most of the vaccinated pigs did not develop any clinical signs of acute erysipelas after the challenge exposure, whereas non vaccinated control pigs died or showed severe generalized erythema with profound depression and anorexia. No differences in the protection against the challenge exposure were observed among the groups. Therefore, the present results suggest that these selected antibiotics would not interfere with the immune effect of the vaccine if given at the usual concentrations used for feed additives. PMID- 11014068 TI - Varying effects of infections with Mycoplasma hyopneumoniae on the weight gain recorded in three different multisource fattening pig herds. AB - Pigs in three specialized fattening herds were studied with respect to the effect of infection with Mycoplasma hyopneumoniae on weight gain. Individual pigs were weighed four times at 4-week intervals during the fattening period and their daily weight gain over the rearing period was calculated. A blood sample was collected on each weighing occasion and analysed for the presence of antibodies to M. hyopneumoniae. The lungs of the principals were inspected at slaughter and the extent of pneumonic lesions was registered by a specially developed technique that has been proven to warrant a high degree of repeatability. No serum antibodies to M. hyopneumoniae were detected in one of the herds, and no pneumonic lesions were recorded at slaughter in that herd. In the other two herds, the prevalence of pigs with serum antibodies to M. hyopneumoniae increased from 6 to 54% and from 31 to 81%, respectively, during the fattening period. The prevalence of pneumonic lesions at slaughter in these herds was higher the later the pigs seroconverted. On the other hand, the extension of the lung lesions tended to be higher among pigs that seroconverted early during the rearing period. Infections with M. hyopneumoniae acquired early during the rearing, presumably strengthened by secondary infections and environmental errors, was found to decrease the daily weight gain of the pigs. However, even non complicated M. hyopneumoniae infections acquired late in the fattening period were associated with reduced daily weight gain. That growth reduction was estimated to be at least 60 g (about 6%) after adjusting for herd, pen, initial weight and sex. PMID- 11014069 TI - Occurrence of chlamydiae in the genital tracts of sows at slaughter and their possible significance for reproductive failure. AB - The aim of this study was to investigate further the role of chlamydiae as pathogens in the genital tracts of sows at slaughter. Genital tracts of 101 randomly selected sows were collected and specimens of genital tract localizations were systematically examined for chlamydiae using immunohistochemistry and PCR. In the genital tracts of 10 sows, Chlamydia psittaci DNA was detected by PCR, and was further typed as 'serotype 1' in nine cases and as avian strain 6 BC in one animal. However, all specimens examined by immunohistochemistry were negative for chlamydiae. Pooled samples of scalding tank water were additionally investigated for 95 animals. Of these samples, 63.2% contained chlamydial DNA, mostly C. trachomatis, and in one sample C. psittaci 'serotype 1'. Although in most cases contamination through influx of faecally contaminated scalding water is a possible reason for the positive PCR results in the genital tract, latent infection cannot be excluded. In conclusion, the results obtained suggest that chlamydiae are of no or only minor importance in the examined group of Swiss breeding sows. Nevertheless, the role and significance of chlamydiae as pathogens in porcine reproductive disorders remain unresolved and require further investigation. PMID- 11014070 TI - HC2000 at Harrogate. Health informatics proceeds with caution. PMID- 11014071 TI - HIPAA 2000 Conference report. HIPAA is heating up. PMID- 11014072 TI - Health information on the Internet: let the viewer beware (caveat viewor). PMID- 11014073 TI - The granting game: Part II. How to love NIH form 398 and get that grant. PMID- 11014074 TI - Buy software or "pay-per-view": the ASP option. PMID- 11014075 TI - The Yale Center for Medical Informatics: clinical, neuro- and genomic informatics. PMID- 11014076 TI - A paradigm shift in healthcare. From disease management to patient-centered systems. PMID- 11014077 TI - New modalities complicate the legal balance. PMID- 11014078 TI - The "bank of health". A model for more useful patient health records. PMID- 11014079 TI - Behavioral health information systems. Evaluating readiness and user acceptance. PMID- 11014080 TI - Aortic dissection. AB - Acute and chronic aortic dissection is described in terms of diagnostics, surgical indications and present day operative treatment. The need for prompt decision making and intervention is highlighted. The results to be expected from surgery are outlined. PMID- 11014081 TI - New criteria for the diagnosis and treatment of atrioventricular node reentrant tachycardia. AB - The authors describe a new elecrocardiographic criterion to diagnose atrioventricular node reentrant tachycardia, review the electrocardiographic and electrophysiological diagnosis of this tachyarrhythmia and analyse the results of catheter ablation of fast versus slow pathway, selecting the fast pathway whenever the induction of atrioventricular node reentrant tachycardia is performed only after pharmacological maneuvers. The high specificity for atrioventricular node reentrant tachycardia of the criterion characterized by the absence of ST-segment depression found in left precordial leads from V4 to V6 suggests the need to include it in electrocardiographic algorithms for the differential diagnosis of tachycardias with a narrow QRS complex. The high primary and final success rates of catheter ablation obtained after a therapy stratification based on induction criteria, emphasize the use of the fast pathway ablation as a first option in selected cases and for the treatment of recurrences. PMID- 11014082 TI - Differential diagnosis of chest pain in an emergency department: the value of homocysteinemia as a diagnostic marker. AB - Only 20 to 45% of patients (pts) admitted to an emergency department (ED) with chest pain (CP) have a cardiac ischemic CP (ICP). Apart from the need for a rapid detection of ICP cases, it is important to avoid discharging patients with false negative ICP. We studied the capability of plasma homocysteine (Hcy) values to improve the differential diagnosis (DD) in patients admitted to an ED with CP. PATIENTS: 125 patients, 80 males, mean age 61 yrs (25-96 yrs), 75 (60%) without prior atherosclerotic cardiovascular disease (ACVD), consecutively admitted to the ED with CP which was not immediately clear. Definitive diagnosis: ICP-58 patients (46%); non-ischemic (non-ICP)--64 patients (54%). METHOD: Hcy was measured on admittance (fasting state not required), by means of a fluorescence polarization enzyme immunoassay (IMx, Abbott); at real conditions, the results were obtained within 2 hours (but not displayed). RESULTS: 1) Hcy (mumol/L) = 10.9 +/- 5.4 (non ICP without prior CDAV); 13.9 +/- 7.7 (ICP). 2) Hcy > or = 15.0 mumol/L in patients without prior ACVD: 5 patients (9%) with non ICP, 8 patients (38%) with ICP--p < 0.01; RR = 2.9 (95% CI = 1.2-7.1); positive value = 62%; negative predictive value = 79%. CONCLUSION: In patients without ACVD, Hcy may contribute to improve DD of equivocal CP, namely: 1) before a normal or non diagnostic ECG and negative ischemia markers, Hcy > or = 15.0 mumol/L will imply additional cardiological investigation in the ED; 2) Hcy < 15.0 increases the liability of a non-ICP diagnosis. PMID- 11014083 TI - Homocysteinemia in the emergency department. PMID- 11014084 TI - Blood pressure in pediatric years (8-13 years old) in the Oporto region. AB - The early detection and treatment of primary and secondary hypertension in children may contribute to the improvement of health later in life. During childhood, the occurrence of arterial hypertension should be understood as a risk situation, even if the clinical signs of the disease are not obvious. Comparatively to other populations, only few epidemiological studies exist in Portugal to be used as clinical references. Thus, the aim of this work was to study the distribution curves of 30, 60 and 90 percentiles of systolic blood pressure (SBP) and diastolic blood pressure (DBP), related to age, height, and weight. To define the "cut off points" of hypertension in our population, we estimated the 95 percentile of SBP and DBP. This is a cross-sectional study. The sample consisted of 474 children (242 boys and 232 girls), aged 8 to 13 years old, randomly selected in 30 different schools of the Oporto region. Weight, height and body mass index were evaluated as anthropometric reference values and blood pressures, SBP and DBP, were measured at rest. The results revealed a gradual increase in blood pressure values with age, height and weight. We found significant correlations (p < or = 0.05; p < or = 0.01) between SBP and DBP to the variables height, weight and blood mass index in both sexes. Multiple regression analysis showed that weight and age explain significantly (p < or = 0.01) the variation of SBP in both sexes. Related to DBP, in males and females, only age accounted significantly for the amount of variance. In conclusion, the present study pointed out the need to associate the SBP and DBP values to the age, weight and height of the subjects, mainly in boys. This work stresses the need for SBP and DBP percentile tables related to age, to weight and to height, for the Portuguese population, particularly for pediatric ages. PMID- 11014085 TI - [Diagnosis of paradoxal and concomitant pulmonary thromboembolism with transesophageal echocardiography: a clinical case]. AB - There is some controversy regarding the indications for transesophageal echocardiography in patients with suspected systemic embolism. The present case report refers to a 65 year old male admitted to the hospital for ischemic acute cerebrovascular accident, which was confirmed by cerebral computerized tomography. A transthoracic echocardiogram was performed showing right atrial and ventricular dilatation. A transesophageal echocardiogram was also performed to exclude thromboembolism and clarify dilatation of the right cavities. Mild spontaneous echocontrast was present in the left atrium without images of thrombus; an interatrial septal aneurysm with patent foramen ovale was found with right to left flow; an image compatible with a very mobile, large, proximal thrombus in the main pulmonary artery was observed. A venous duplex scan was performed, demonstrating venous thrombosis in the right popliteal and femoral veins. Pulmonary arteriography showed a large thrombus in the right pulmonary branch extending to the median lobe and a smaller thrombus in the left pulmonary branch. Apparently, the patient had no predisposing factors for thromboembolism. Full-dose heparin was started followed by oral anticoagulation. An inferior vena cava filter was implanted. At hospital discharge the pulmonary thrombus had disappeared and the right cardiac cavities had returned to normal size. The interatrial aneurysm had disappeared and foramen ovale was no longer patent. After 36 months of clinical follow up on oral anticoagulation, the patient remains asymptomatic without neurological sequelae nor respiratory distress. PMID- 11014086 TI - Severe mitral regurgitation due to a leak of a mitral ring--an echocardiographic view. PMID- 11014087 TI - Myocarditis due to influenza virus complicated by intravascular coagulopathy. PMID- 11014088 TI - Cardiopulmonary resuscitation by chest compression alone or with mouth-to-mouth ventilation. PMID- 11014089 TI - [Research in axillary lymphatic metastasis...which techniques for which material? What is at stake?]. PMID- 11014090 TI - [Lymph node micrometastasis? Reflections on their definition, demonstration, significance and conclusions to be drawn]. PMID- 11014091 TI - [The place of surgery in the treatment of advanced localized, recurrent and metastatic breast cancer]. AB - In the multidisciplinary treatment of locally advanced and metastatic breast cancer, aggressive surgical options can be chosen in selected cases. They may allow: survival to be prolonged by the resection of metastases (liver, ovary, lung), symptomatic treatment (bone pain, local recurrence, infiltration of the chest wall), prevention of potentially disabling complications (pathologic fractures, medullary compression), exclusion of another tumoural or non tumoural diseases. The decision to perform surgery has to be discussed between the surgeon and the oncologist so as to optimise its timing. Surgical treatment can follow induction therapy or can precede chemo- or hormonotherapy. PMID- 11014092 TI - [Sentinel lymph node biopsy in breast cancer: the Lausanne experience]. AB - A total of 40 patients (mean age 51 yrs; 36-89 yrs) with clinically T1-T2(< 3 cm)N0M0 breast cancer underwent sentinel node (SN) mapping with radioactive tracer (99mTc) injection only in 21 patients, with Patent blue V in 1 patient, or with both techniques in 19 patients. The preoperative injection of 99mTc (20-40 MBq) was followed by lymphoscintigraphy. A handheld gamma probe was used to detect the SN in the operative room. A lumpectomy and an axillary dissection were performed in all the patients. SNs could be identified in 39/40 patients, resulting in a sensitivity of 98%. Successful localization of the SNs was accomplished by isotope only in 19/20 patients, by blue dye only in 1/1 patient, and by both methods in 19/19 patients; in 2 of these 19 patients, SNs were identified by blue dye only. Axillary metastases were found in 12/40 patients (30%), the SN being the only nodal metastasis in 8/12 patients (75%). Six of these 12 patients (50%) had only evidence of micrometastasis. Negative SNs on serial sections stained with hematoxylin-eosin (H&E) were evaluated with cytokeratin immunostain (C11). In all cases of negative SNs the remaining axillary nodes were also free of tumor, resulting in a negative predictive value of 100%. We conclude that SN mapping is a highly accurate method for staging the axillary node status in breast cancer patients. Optimal localization is achieved by the combination of injection of 99mTc-colloid and blue dye as evidence by the cases of positive SN identified by only one of both methods. PMID- 11014093 TI - [Aromatase inhibitors in the treatment of breast cancer]. PMID- 11014094 TI - [Organized screening for breast cancer: the Vaud experience]. PMID- 11014095 TI - [Chemoprevention of breast cancer: a hope justified?]. AB - Chemoprevention of breast cancer, a dream? In one American study (National Surgical Adjuvant Breast and Bowel Project, NSABP-P1) tamoxifen decreased the incidence of invasive and non-invasive breast cancer, whereas in two English and Italian studies, it did not reduce this risk. Differences in the study population for the trials may underly these conflicting findings. New drugs, the SERMs, may be more promising. PMID- 11014096 TI - [What's new in hormone replacement therapy for postmenopausal women? I. Advantages of hormone replacement therapy (HRT)]. AB - Different cohort studies have shown that HRT decreases the risk of cardio vascular (C-V) disease and the risk of bone fracture by 30 to 50%. The only controlled study (HERS study) did not show any benefit of HRT with estradiol and medroxyprogesterone (MPG) in secondary prevention. The beneficial effect of estrogens on coronary dilatation and on HDL cholesterol could be attenuated by some progestogens such as MPG but not by nomegestrol acetate. In this framework, the comparative metabolic effects of different progestogens and tibolone are described in this article. The effects of estrogens on mood and of androgens on libido are discussed. The preventive effect of estrogens on osteoporosis and on Alzheimer disease is compared to other nonhormonal treatments. PMID- 11014097 TI - [What's new in hormone replacement therapy of postmenopausal women? II. Risks of hormone replacement therapy (HRT)]. AB - A 50 year old woman has a 10% lifetime risk of developing a breast cancer. Depending on the duration of the treatment, HRT can increase this risk by 30 to 45%. The risk of endometrial cancer, which affects 2.3% of women, is increased even if sequential progestogens are given together with estrogens. The risk of venous thrombosis is increased 3 times. The occurrence of ictus is not modified by HRT. On the other hand estrogens may prevent the abdominal deposit of fat. The cancer risks associated with HRT must be balanced against their protective effects on cardio-vascular (C-V)diseases. In untreated women, mortality due to C V disease is 39% whereas mortality due to breast cancer is 3% and only 0.3% for endometrial cancer. This article discusses also the possibility of HRT and of non hormonal treatments in patients with previous breast cancer. PMID- 11014098 TI - [Interleukin-1 inhibitors: a new alternative for the treatment of rheumatoid polyarthritis]. PMID- 11014099 TI - [Positron emission tomography (PET) with 18F-fluorodeoxyglucose (18F-FDG) in ORL and pulmonary oncology]. PMID- 11014100 TI - [St. John's wort--scientific tests. Report of an expert workshop in Berlin]. PMID- 11014101 TI - [Disability evaluation: support from the model of complexity]. PMID- 11014102 TI - [Image of the month. Renal vein thrombosis in the neonatal period]. PMID- 11014103 TI - [How I treat...decubitus ulcers surgically]. AB - Although pressure sores occur frequently during the evolution of paraplegic patients, they can present in patients confined to bed or in casts. Ischiatic, sacral and trochanteric sores are the most frequent. For stade III & IV lesions surgical excision followed by a coverage with healty tissue remain the only right principle. Usually, flap transfert meats these requirements. However, the selection of the surgical procedure and the post-op care are the main factors for a successful treatment. PMID- 11014104 TI - [Clinical case of the month. Abdominal wall hematoma and macroscopic hematuria in a 95 year-old patient]. AB - We report the case of a 95 year-old female patient who died from hypovolemic shock caused by severe hematuria and abdominal hemorrhage. The necropsy demonstrated a primary signet-ring cell adenocarcinoma of the urinary bladder. The diagnosis, prognosis and histogenesis of this uncommon tumor are discussed. PMID- 11014105 TI - [Medical-legal aspects in sexual assault matters]. AB - The examination of a victim of sexual acts represents an important activity in forensic medicine, so quantitatively that by his contribution to truth demonstration. Either ponctual or repeated facts, this investigation, required by judicial authorities, necessitates a big availability towards victims, an anatomical thorough knowledge in particular of various aspects of hymen as well as samples of high quality. This article lays emphasis on basic points for the good accomplishment of this forensic examination. PMID- 11014106 TI - [Bronchial morphologic modification in asthma]. AB - Asthma is an inflammatory disease of the airways clinically characterised by recurrent bronchial obstructions at least partially reversible. Recent epidemiologic data suggest that asthmatics have an increased rate of decrease of their expiratory volumes during life. This irreversible lung function impairment is associated with fundamental structural changes of the bronchial wall in terms of conjunctive tissue and smooth muscle composition. We describe these changes and explore the different mechanisms proposed to explain these structural modifications. We also review their consequences in terms of bronchial physiology and their potential influence on bronchial hyperresponsiveness. PMID- 11014107 TI - [Hydroxyurea and HIV infection]. AB - Hydroxyurea is an anticancerous product, used recently in the treatment of HIV-1 infection thanks to its inhibitory action in viral replication, potentialization of the nucleosides activity (particularly ddI or didanosine) and its cytostatic properties on CD4 and CD8 lymphocytes. Many studies showed its efficiency, as further drug, in initial regimen of a tritherapy (containing ddI) and salvage therapy. The dosage of 500 mg bid seems tolerated well by adults, and 20 mg/kg by children. Long-term tolerance remains unknown. With ddI, it could be proposed in developing countries. PMID- 11014108 TI - [The IGF system: summary and recent data]. AB - We are entitled to state that our knowledge about the IGF system has literally exploded in the last years. Having been considered for some time merely as trophic and mitogenic factors, the IGFs now appear as molecules essential for the differentiation of many cell types, and even more so, as powerful protective agents for the nervous and the cardiovascular systems. However, these properties so beneficial in normal physiological conditions, are subverted by the cancerous cells who use them to extend their life span and resist therapy. The IGFs did not live up to expectations in the treatment of diabetes; however, today their capacity to improve the condition of patients suffering from severe neurological, renal or muscle diseases is tested. The IGF system might also be targetted by the anticancer treatments. In the following paper we have briefly summarized our knowledge on the IGF system, and presented in more detail the recent data. PMID- 11014109 TI - [250 years ago "L'Homme machine" was published. La Mettrie (1709-1753). The origins of medical materialism]. AB - Some 250 years ago, the publication of "L'Homme machine" established the fact that human body and brain functioning was to be approached by human study just as like as physics or chemistry. PMID- 11014110 TI - [How I investigate...fetal pathology. Technique and value of the fetal and placental examination and the perinatal autopsy]. AB - Fetal pathology is a recent field in pathological anatomy, knowing at the present time an important technical and theoretical development. However, it is still insufficiently used to resolve cases of fetal death in utero, or peripartum or perinatal. In this paper, we describe the basics and the techniques used during the fetal examination. We underline the values of these techniques for the understanding of failures of pregnancy and for orientation of the genetical advice for future pregnancies. PMID- 11014111 TI - [Pharma-clinics. The drug of the month. Rofecoxib (Vioxx)]. AB - Rofecoxib (Vioxx, Merck Sharp & Dohme) is a potent and selective inhibitor of the COX-2 isoform of cyclooxygenase which is used as a nonsteroidal anti-inflammatory drug (NSAID). It is indicated in the symptomatic relief of pain due to osteoarthritis. The initial oral dosage of rofecoxib is 12.5 mg once daily in adults, and this dose may be increased up to a maximal dosage of 25 mg once daily if necessary. Its clinical efficacy seems to be similar to that of other NSAIDs at maximal recommended dosages, but its safety profile, especially gastrointestinal tolerance, is much better because of the COX-2 selectivity. Ongoing clinical trials are performed in patients with rheumatoid arthritis. PMID- 11014112 TI - [Clinical study of the month. The ALLHAT study: a recommendation for prudence in the use of alpha-blockers for the treatment of hypertension]. PMID- 11014113 TI - [What's new in the domain of obesity?]. AB - We will briefly comment on some interesting topics presented at the 10th European Congress on Obesity (ECO, Antwerp, Belgium, 24-27 May 2000). Special attention has been focused upon: 1) the multi-endocrine function of the adipocyte cell; 2) the role of gene-environment interactions; 3) the regulatory mechanisms of energy balance; 4) the neuroendocrine hypothesis for the development of visceral adiposity; 5) the numerous complications associated with obesity, more particularly type 2 diabetes; and, finally, 6) obesity prevention, especially in childhood, and new pharmacological perspectives for the treatment of obesity. PMID- 11014114 TI - [Rationing in health care: the elderly as victims?]. PMID- 11014115 TI - [Rationing: the royal path in health care?]. PMID- 11014116 TI - [Ethically fair accessibility in health care]. PMID- 11014117 TI - [Political considerations in discussing health care rationing]. PMID- 11014118 TI - [Rationing in the hospital: realities and decision pathways]. PMID- 11014119 TI - [Age as a rationing criterium?]. PMID- 11014120 TI - [Allocation of donated organs for elderly patients]. PMID- 11014121 TI - [Rational service allocation instead of threatened rationing of requested services for the elderly. The example of neurorehabilitation]. PMID- 11014122 TI - [Dementia, a rational rationing criterium?]. PMID- 11014123 TI - [Decision making in dementia--from the viewpoint of affected relatives]. PMID- 11014124 TI - [Attitude of seniors to rationing in health care--before and after a lecture series]. AB - Prior to the first lecture in the public series of lectures entitled "Rationing of Health Care: Are Elderly the Victims?" and before a lecture given to a Senior Citizens Association on the same subject, all participants were polled. The polling was repeated at the end of the lecture series. The initial participation totalled 69 with two-thirds women, average age 64 years (50% were from the health care or social welfare sector), and 57 retired business people (50% women). At the end, there were 45 listeners (average age 65 years), with 30 taking part in both surveys. The majority believed that rationing is already taking place, only 1/6 rejected such measures in principle. When asked, nearly all those surveyed were in favor of quality of life after treatment and life expectancy as criteria for the right to medical services. Personal criteria, such as nationality, social role, self-related causes and calendar age were rejected by the majority, although less clearly by the business people than by the general participants. This rejection became more marked after the lecture series. Those surveyed gave various specific judgements on decisions about the complex allocation of resources and favored decision-making by patients themselves unless they were senile. A large minority were opposed to very elderly patients making decisions themselves, even if they were not senile. CONCLUSION: The elderly persons polled demand that all rationalization measures be exploited before rationing is implemented. Rationing criteria must be of a universal nature and borne by the general public. PMID- 11014125 TI - [Proposals for rationing in public health from the viewpoint of the elderly]. PMID- 11014126 TI - [What is your diagnosis? Large-cell bronchial carcinoma cT3 N2-3 M0]. PMID- 11014127 TI - [Standard hepatitis B therapy]. AB - Standard treatment of chronic hepatitis B virus infection includes the use of interferon-a and antiviral nucleoside analogues, alternatively. In general, in patients with hepatic inflammatory activity, low viral titers and HBe positive wild-type infection interferon leads to a long-term seroconversion rate of about 30-50%. However in patients with advanced liver disease, in interferon non responders and HBe minus mutants HBV carriers nucleoside analogues like lamivudine represents a new and clinically proven therapeutical option. However, the longterm outcome and the problem of the emergence of HBV polymerase gene mutants has not been completely understood. PMID- 11014128 TI - ["Ultrasonographically detected liver lesion"--how to proceed?]. AB - Focal liver lesions found incidentally in ultrasound are predominantly uncomplicated cysts, cavernous hemangiomas or typical metastases. In most of these cases a reliable diagnosis can be made. Atypical findings, however, always are suspicious being malignant. In this particular situation imaging methods, such as CT, NMR or scintigraphy, should be applied. Despite of this a final diagnosis may not be found in all cases. An ultrasound-guided fine-needle biopsy is then required. Biopsy during laparotomy might be the procedure of choice in some situations. PMID- 11014129 TI - [Therapy of pancreatic and bile duct tumors: value of radiotherapy and photodynamic therapy]. AB - Prognosis of pancreatic cancer and bile duct cancer is extraordinary poor. Despite of the improvement in diagnostic procedures and the development of more radical resectional procedures prolongation of survival could not have been achieved yet. At the time of diagnosis only 20-30% of patients are suitable for resection, thus the value of non resectional methods for palliation has to be determined. Despite of numerous efforts to increase the local radiation dosage by means of distinguished radiation techniques none of these procedures could achieve an unequivocal prolongation of survival. After promising preliminary results of intraoperative radiotherapy (IORT) in pancreatic cancer following studies could not show a survival benefit for IORT allone or in combination with extracorporal radiotherapy (EBRT). EBRT is indicated at best for local pain control and is limited by the high radiation sensitivity of the adjacent organs. Intracorporal radiotherapy or brachytherapy with iridium 129-labelled wires leeds to local, intraluminal tumour control, but has also no influence on survival time. Up to now, photodynamic therapy (PDT) of pancreatic cancer must be characterized as highly experimental. But PDT can contribute to local tumour control. Preliminary data of photodynamic therapy in bile duct cancer show good results concerning reduction of cholestasis, improvement of life quality and even prolongation of survival time. The value of these preliminary results is being proved at present in controlled studies in comparison to mere endoprosthetic supply at a few centers. PMID- 11014130 TI - [Risk groups for pancreatic and bile duct carcinomas]. AB - Biliopancreatic carcinoma has a poor prognosis since the diagnosis of the tumor occurs late when advanced disease is present. The identification of potential causes and earlier diagnosis are needed to prevent the disease or identify it early enough to improve survival. The main risk factors for pancreatic cancer include advanced age, cigarette smoking, high-fat diet, diabetes mellitus, chronic pancreatitis (especially hereditary pancreatitis) and a positive family history of pancreatic cancer. The most important etiologic factor for the development of gallbladder cancer is gallstone disease. Patients with anatomic abnormalities and chronic inflammatory conditions (primary sclerosing cholangitis, infections with parasites) have an increased incidence of bile duct cancers. Several new and promising imaging techniques have recently become available and our understanding of the mechanisms of carcinogenesis are growing rapidly. However, there is currently no effective screening strategy applicable and it is unknown when to begin screening. For pancreatic cancer, reduction of risk is likely to occur with avoidance of smoking and promotion of healthful diets. Cholecystectomy rates have increased since the introduction of new laparoscopic techniques and will eventually reduce the incidence of gallbladder cancer. Improved imaging techniques, the identification of new genes and a better definition of genetic alterations that characterize preinvasive lesions will hopefully allow to develop sensitive and specific technologies to screen and to detect early biliopancreatic cancer for even premalignant lesions to improve the mostly fatal prognosis if this tumor. PMID- 11014132 TI - [Dengue fever in Denmark]. PMID- 11014131 TI - [Diabetic nephropathy. A modifiable risk factor]. PMID- 11014133 TI - [Risk factors in the progression of diabetic nephropathies]. AB - Diabetic nephropathy is a serious long-term complication of diabetes characterized by persistent albuminuria (> 300 mg/24 h or 200 micrograms/min) associated with a decline in GFR, increasing arterial blood pressure, and high risk of cardiovascular morbidity and mortality. Diabetic nephropathy has become the leading cause of end stage renal disease in Europe, USA and Japan. There is a great inter-individual variation in the deterioration in GFR explained by genetic and non-genetic progression promoters (risk factors for losing GFR). In particular arterial blood pressure, glycaemic control, and albuminuria act as non genetic promoters of progression. These factors are potentially modifiable. Genetic promoters of progression have also been identified, mainly the angiotensin converting enzyme insertion/deletion polymorphism. Knowledge of progression promoters can identify patients with a poor prognosis and direct therapy against modifiable risk factors, in particular hypertension and hyperglycaemia. PMID- 11014135 TI - [Hereditary dystonias]. AB - Dystonia is a heterogeneous, neurological disease characterized by involuntary, sustained muscle contractions, frequently causing twisting and repetitive movements or abnormal postures. The patients are often difficult to diagnose, and the treatment is almost always only symptomatic. It is believed that about 75% of all patients with dystonia have primary dystonia, and 25-85% of these are hereditary. Seven gene loci for autosomal, dominant inherited dystonia and two for X-linked, recessive inherited dystonia are known at present, but the underlying genes are known only for DYT1 and DYT5. Testing is possible for these two in Denmark. Growing molecular genetic knowledge will lead to earlier and correct diagnosing, including prognosis, and may elucidate the pathogenesis, making better treatment possible. PMID- 11014134 TI - [Dengue fever and dengue hemorrhagic fever]. AB - Dengue virus is transmitted by mosquitoes and causes dengue fever/dengue haemorrhagic fever throughout the tropical areas of the world. There is an increasing incidence of dengue infections. Because of increasing travel activity, infection among Danes travelling abroad as well as imported cases are expected to be seen more frequently. In this review we describe the clinical manifestations, diagnosis, pathogenesis, treatment and prevention of the disease. PMID- 11014136 TI - [Bite wounds and infection prophylaxis. Evidence-based therapeutic recommendations]. AB - Infections are common complications of bite wounds, changing in severity from local cellulitis to life-threatening septicaemia. Treatment recommendations are often contradictory and rarely reflect evidence-based medicine. Risk factors associated with higher rates of infection include localization on the hands, puncture wounds and age older than 50 years. The literature is critically reviewed for evidence-based treatment of bite wounds. The results are discussed with respect to common practice in Denmark. PMID- 11014137 TI - [Dengue fever among 44 Danish travellers investigated in the department of epidemics at the Rigshospitalet during 1988-1998]. AB - Dengue fever is a major cause of febrile illness in the tropics, and we describe 44 patients with dengue fever seen at Rigshospitalet, Copenhagen from 1988-98. A worldwide increase in transmission of dengue fever was reflected in the number of patients seen in 1997-1998. All patients had fever and headache, and were biochemically characterised by thrombocytopenia, leucopenia, increased levels of alanine aminotransferase and a rise in haematocrit. One patient had dengue haemorrhagic fever, and two patients exhibited unusual reactions to the infection, one in the form of extended febrile neutropenia complicated by an episode of of E. coli septicaemia, and one patient developed progressive paralysis of both legs, and remains partially paretic. PMID- 11014138 TI - [The amount of time spent in processing of 40 cases involving involuntary commitment under the legislation of 1989 in a Danish county 1990-1994]. AB - This investigation examines forty cases dealing with compulsory admission or detention which have been presented at the Local Magistrates Court in a Danish county under the Danish Mental Health Legislation of 1989 during a five year period from 1990-1994. The amount of time spent in processing the cases by the psychiatric department, the Forensic Medical Council and the magistrate was registered. The results show a written psychiatric statement from the department is available after seven to ten days. Five weeks pass when the case is presented to the Forensic Medical Council, (four weeks with the Council and one week with the magistrate). The importance of obtaining statements from the Forensic Medical Council and the time spent in considering the cases with reference to the Justice Department's survey of the Danish Mental Health Legislation of 1989 is discussed. The investigation concludes that despite the resulting prolonged case evaluation it is recommended that cases involving detainment are brought before the Forensic Medical Council. PMID- 11014140 TI - [Two years of experiences with a county clinic specialized for breast diseases]. PMID- 11014139 TI - [Stress and fertility. A follow-up study among couples planning the first pregnancy]. AB - Mental distress has often been suggested as a cause of unexplained infertility. However, the causal direction may well be from infertility to distress, and prospective data are needed. We therefore followed 393 couples, who were planning their first pregnancy, with prospective collection of information on distress from termination of birth control until pregnancy for a maximum of six menstrual cycles. The analyses included 1,475 menstrual cycles, and mental distress was measured in each cycle by the General Health Questionnaire (GHQ). Urine samples from each period of vaginal bleeding were analyzed for human chorionic gonadotrophic hormone, indicating early embryonal loss. We found that for cycles with the highest distress score (GHQ score above the 80 percentile) the probability of conception per cycle was 12.8% compared to 16.5% in other cycles (adjusted OR 0.6; 95% CI 0.4-1.0). The effect of distress was almost exclusively found among women with long menstrual cycles (OR 0.1; 95% CI 0.01-0.4 and OR 0.9; 0.5-1.4 for women with cycles > or = 35 and < 35 days, respectively). An increased incidence of early embryonal loss was also found among highly distressed women with long cycles, but was based on a small number of observations. It is concluded that psychological distress may be a risk factor for reduced fertility in women with long menstrual cycles. PMID- 11014141 TI - [Spontaneous infrapatellar tendon rupture in a patient with systemic lupus erythematosus]. AB - A case is described of a 33-year old woman with systemic lupus erythematosus (SLE) in longterm treatment with corticosteroids who experienced spontaneous rupture of the left patellar tendon. A comparative study of 28 previously reported cases of SLE patients with spontaneous tendon rupture in weight bearing joints is performed. It is suggested that renal disease may be an etiological factor for spontaneous tendon rupture in patients with systemic lupus erythematosus. PMID- 11014142 TI - [Dengue fever]. AB - Dengue virus infections have increased globally. This results in an increased incidence of patients with dengue fever in Denmark. In order to draw attention to this diagnosis among febrile persons returning from a stay in tropical and subtropical areas, we present two cases of dengue virus infection and describe the symptoms and diagnostic possibilities. Dengue Haemorrhagic Fever (DHF) is supposed to occur only in persons who have previously had dengue virus infection and who acquire a second infection with dengue virus. PMID- 11014143 TI - [A complicated course of Salmonella gastroenteritis during antihypertensive treatment]. AB - Although Salmonella gastroenteritis usually is a fairly mild and self-limiting disease, serious complications are sometimes seen. We present a case that was further complicated by treatment with enalapril. We recommend discontinuation of treatment with ACE inhibitors in patients with any kind of ongoing gastroenteritis with fluid loss in order to prevent a potentially serious outcome. PMID- 11014144 TI - [Breakthrough for gene therapy]. PMID- 11014145 TI - [Healthy soul]. PMID- 11014146 TI - [Screening for lung cancer--to prevent?]. PMID- 11014147 TI - [Helicobacter pylori diagnostics--how, when and by whom?]. PMID- 11014149 TI - Bridging the Taiwan Strait PMID- 11014148 TI - [Genetic mapping]. PMID- 11014150 TI - Reviews for reviews' sake PMID- 11014152 TI - UK advised to step up asteroid hunt PMID- 11014151 TI - Doubts grow over discovery of fossilized 'dinosaur heart'. PMID- 11014154 TI - Hunt for Higgs particle wins time for CERN collider PMID- 11014153 TI - World Bank is urged to give greater priority to science PMID- 11014155 TI - European space panel picks new priorities PMID- 11014156 TI - NASA reaches out to universities. PMID- 11014158 TI - Case of the stolen Enigma machine takes cryptic turn PMID- 11014157 TI - Germline gene therapy needs tight control, says US panel. PMID- 11014159 TI - Japan opens access to mouse cDNA data...and to report on strengths and weaknesses of genomics centre. PMID- 11014160 TI - A chemistry set for life. PMID- 11014161 TI - Allowing gene patents could be an expensive mistake for the US. PMID- 11014162 TI - No room on the carousel for meeting of like minds PMID- 11014163 TI - Standardized addresses would make web easier. PMID- 11014165 TI - Sky was not the limit for music-loving Herschel PMID- 11014164 TI - The Durban Declaration is not accepted by all. PMID- 11014166 TI - Good and bad science in US schools. PMID- 11014168 TI - The Great Goodbye PMID- 11014167 TI - Down to earth. PMID- 11014169 TI - Electrochemistry. A moving oxygen story PMID- 11014170 TI - Molecular biology. Small subunit, big science. PMID- 11014171 TI - Galactic rotation in real time PMID- 11014172 TI - Cancer. A radical approach to treatment. PMID- 11014173 TI - Ocean biogeochemistry. Calcification and CO2. PMID- 11014174 TI - AIDS. Escape from the immune system. PMID- 11014175 TI - Developmental biology. Post-expressionist flies. PMID- 11014176 TI - Elvin A. Kabat (1914-2000). PMID- 11014177 TI - Membrane changes during hibernation. PMID- 11014178 TI - Winter torpor in a large bird. PMID- 11014179 TI - Cloning of mice to six generations. PMID- 11014180 TI - Total silencing by intron-spliced hairpin RNAs. PMID- 11014181 TI - Loosening of plant cell walls by expansins. AB - Plant cell walls are the starting materials for many commercial products, from lumber, paper and textiles to thickeners, films and explosives. The cell wall is secreted by each cell in the plant body, forming a thin fibreglass-like network with remarkable strength and flexibility. During growth, plant cells secrete a protein called expansin, which unlocks the network of wall polysaccharides, permitting turgor-driven cell enlargement. Germinating grass pollen also secretes an unusual expansin that loosens maternal cell walls to aid penetration of the stigma by the pollen tube. Expansin's action has puzzling implications for plant cell-wall structure. The recent explosion of gene sequences and expression data has given new hints of additional biological functions for expansins. PMID- 11014182 TI - Structure of the 30S ribosomal subunit. AB - Genetic information encoded in messenger RNA is translated into protein by the ribosome, which is a large nucleoprotein complex comprising two subunits, denoted 30S and 50S in bacteria. Here we report the crystal structure of the 30S subunit from Thermus thermophilus, refined to 3 A resolution. The final atomic model rationalizes over four decades of biochemical data on the ribosome, and provides a wealth of information about RNA and protein structure, protein-RNA interactions and ribosome assembly. It is also a structural basis for analysis of the functions of the 30S subunit, such as decoding, and for understanding the action of antibiotics. The structure will facilitate the interpretation in molecular terms of lower resolution structural data on several functional states of the ribosome from electron microscopy and crystallography. PMID- 11014183 TI - Functional insights from the structure of the 30S ribosomal subunit and its interactions with antibiotics. AB - The 30S ribosomal subunit has two primary functions in protein synthesis. It discriminates against aminoacyl transfer RNAs that do not match the codon of messenger RNA, thereby ensuring accuracy in translation of the genetic message in a process called decoding. Also, it works with the 50S subunit to move the tRNAs and associated mRNA by precisely one codon, in a process called translocation. Here we describe the functional implications of the high-resolution 30S crystal structure presented in the accompanying paper, and infer details of the interactions between the 30S subunit and its tRNA and mRNA ligands. We also describe the crystal structure of the 30S subunit complexed with the antibiotics paromomycin, streptomycin and spectinomycin, which interfere with decoding and translocation. This work reveals the structural basis for the action of these antibiotics, and leads to a model for the role of the universally conserved 16S RNA residues A1492 and A1493 in the decoding process. PMID- 11014184 TI - The accelerations of stars orbiting the Milky Way's central black hole AB - Recent measurements of the velocities of stars near the centre of the Milky Way have provided the strongest evidence for the presence of a supermassive black hole in a galaxy, but the observational uncertainties poorly constrain many of the black hole's properties. Determining the accelerations of stars in their orbits around the centre provides much more precise information about the position and mass of the black hole. Here we report measurements of the accelerations of three stars located approximately 0.005 pc (projected on the sky) from the central radio source Sagittarius A* (Sgr A*); these accelerations are comparable to those experienced by the Earth as it orbits the Sun. These data increase the inferred minimum mass density in the central region of the Galaxy by an order of magnitude relative to previous results, and localize the dark mass to within 0.05 +/- 0.04 arcsec of the nominal position of Sgr A*. In addition, the orbital period of one of the observed stars could be as short as 15 years, allowing us the opportunity in the near future to observe an entire period. PMID- 11014185 TI - Onset of antiferromagnetism in heavy-fermion metals AB - There are two main theoretical descriptions of antiferromagnets. The first arises from atomic physics, which predicts that atoms with unpaired electrons develop magnetic moments. In a solid, the coupling between moments on nearby ions then yields antiferromagnetic order at low temperatures. The second description, based on the physics of electron fluids or 'Fermi liquids' states that Coulomb interactions can drive the fluid to adopt a more stable configuration by developing a spin density wave. It is at present unknown which view is appropriate at a 'quantum critical point' where the antiferromagnetic transition temperature vanishes. Here we report neutron scattering and bulk magnetometry measurements of the metal CeCu(6-x)Au(x), which allow us to discriminate between the two models. We find evidence for an atomically local contribution to the magnetic correlations which develops at the critical gold concentration (x(c) = 0.1), corresponding to a magnetic ordering temperature of zero. This contribution implies that a Fermi-liquid-destroying spin-localizing transition, unanticipated from the spin density wave description, coincides with the antiferromagnetic quantum critical point. PMID- 11014186 TI - Detection of geometric phases in superconducting nanocircuits AB - When a quantum-mechanical system undergoes an adiabatic cyclic evolution, it acquires a geometrical phase factor' in addition to the dynamical one; this effect has been demonstrated in a variety of microscopic systems. Advances in nanotechnology should enable the laws of quantum dynamics to be tested at the macroscopic level, by providing controllable artificial two-level systems (for example, in quantum dots and superconducting devices). Here we propose an experimental method to detect geometric phases in a superconducting device. The setup is a Josephson junction nanocircuit consisting of a superconducting electron box. We discuss how interferometry based on geometrical phases may be realized, and show how the effect may be applied to the design of gates for quantum computation. PMID- 11014187 TI - Ordering and self-organization in nanocrystalline silicon AB - The spontaneous formation of organized nanocrystals in semiconductors has been observed during heteroepitaxial growth and chemical synthesis. The ability to fabricate size-controlled silicon nanocrystals encapsulated by insulating SiO2 would be of significant interest to the microelectronics industry. But reproducible manufacture of such crystals is hampered by the amorphous nature of SiO2 and the differing thermal expansion coefficients of the two materials. Previous attempts to fabricate Si nanocrystals failed to achieve control over their shape and crystallographic orientation, the latter property being important in systems such as Si quantum dots. Here we report the self-organization of Si nanocrystals larger than 80 A into brick-shaped crystallites oriented along the (111) crystallographic direction. The nanocrystals are formed by the solid-phase crystallization of nanometre-thick layers of amorphous Si confined between SiO2 layers. The shape and orientation of the crystallites results in relatively narrow photoluminescence, whereas isotropic particles produce qualitatively different, broad light emission. Our results should aid the development of maskless, reproducible Si nanofabrication techniques. PMID- 11014188 TI - Direct electrochemical reduction of titanium dioxide to titanium in molten calcium chloride AB - Many reactive metals are difficult to prepare in pure form without complicated and expensive procedures. Although titanium has many desirable properties (it is light, strong and corrosion-resistant), its use has been restricted because of its high processing cost. In the current pyrometallurgical process--the Kroll process--the titanium minerals rutile and ilmenite are carbochlorinated to remove oxygen, iron and other impurities, producing a TiCl4 vapour. This is then reduced to titanium metal by magnesium metal; the by-product MgCl2 is removed by vacuum distillation. The prediction that this process would be replaced by an electrochemical route has not been fulfilled; attempts involving the electro deposition of titanium from ionic solutions have been hampered by difficulties in eliminating the redox cycling of multivalent titanium ions and in handling very reactive dendritic products. Here we report an electrochemical method for the direct reduction of solid TiO2, in which the oxygen is ionized, dissolved in a molten salt and discharged at the anode, leaving pure titanium at the cathode. The simplicity and rapidity of this process compared to conventional routes should result in reduced production costs and the approach should be applicable to a wide range of metal oxides. PMID- 11014189 TI - Reduced calcification of marine plankton in response to increased atmospheric CO2. AB - The formation of calcareous skeletons by marine planktonic organisms and their subsequent sinking to depth generates a continuous rain of calcium carbonate to the deep ocean and underlying sediments. This is important in regulating marine carbon cycling and ocean-atmosphere CO2 exchange. The present rise in atmospheric CO2 levels causes significant changes in surface ocean pH and carbonate chemistry. Such changes have been shown to slow down calcification in corals and coralline macroalgae, but the majority of marine calcification occurs in planktonic organisms. Here we report reduced calcite production at increased CO2 concentrations in monospecific cultures of two dominant marine calcifying phytoplankton species, the coccolithophorids Emiliania huxleyi and Gephyrocapsa oceanica. This was accompanied by an increased proportion of malformed coccoliths and incomplete coccospheres. Diminished calcification led to a reduction in the ratio of calcite precipitation to organic matter production. Similar results were obtained in incubations of natural plankton assemblages from the north Pacific ocean when exposed to experimentally elevated CO2 levels. We suggest that the progressive increase in atmospheric CO2 concentrations may therefore slow down the production of calcium carbonate in the surface ocean. As the process of calcification releases CO2 to the atmosphere, the response observed here could potentially act as a negative feedback on atmospheric CO2 levels. PMID- 11014190 TI - Increased dissolved oxygen in Pacific intermediate waters due to lower rates of carbon oxidation in sediments. AB - Concentrations of dissolved oxygen in the ocean seem to correlate well with climate instabilities over the past 100,000 years. For example, the concentration of dissolved oxygen in Pacific intermediate waters was considerably higher during Pleistocene glacial periods than it is today. This has been inferred from the presence of bioturbated sediments, implying that oxygen levels were sufficient for burrowing organisms to live. Today, basins in the northeastern Pacific Ocean are floored by laminated sediments implying lower oxygen levels, which may be explained by reduced ventilation. Here we report a recent return to bioturbated sediments in the northeastern Pacific Ocean since the late 1970s. From the carbon isotope composition of benthic foraminifers living in the sediment, we infer a twofold decrease in the carbon oxidation rate occurring within sediments, equivalent to an increase in dissolved oxygen concentration of 15-20 micromoles per litre. These changes, at the edges of the Santa Barbara, Santa Monica and Alfonso basins, are coincident with a change in North Pacific climate which has reduced upwelling by 20-30% and increased sea surface temperatures by 1.5-3 degrees C. This suggests that climate effects on surface productivity, reducing the supply organic matter to sediments, may have had a greater effect on benthic oxygen levels than changes in ocean circulation patterns. PMID- 11014191 TI - Variance in ecological consumer-resource interactions. AB - Food-web models use the effect size of trophic interactions to predict consumer resource dynamics. These models anticipate that strong effects of consumers increase spatial and temporal variability in abundance of species, whereas weak effects dampen fluctuations. Empirical evidence indicates that opposite patterns may occur in natural assemblages. Here I show that spatial variance in the distribution of resource populations is sensitive to changes in the variance of the trophic interaction, in addition to the mean effect of consumers, relative to other causes of spatial variability. Simulations indicate that both strong and weak direct effects of consumers can promote spatial variability in abundance of resources, but only trophic interactions with a large mean effect size can reduce variation. Predictions of the model agree with the results of repeated field experiments and are consistent with data from published consumer-resource interactions, proving to be robust across widely varying environmental conditions and species' life histories. Thus, food-web models that embody variance in trophic interactions may have increased capacity to explain the wide range of effects of consumers documented in empirical studies. PMID- 11014192 TI - Body image as a visuomotor transformation device revealed in adaptation to reversed vision. AB - People adapt with remarkable flexibility to reversal of the visual field caused by prism spectacles. With sufficient time, this adaptation restores visually guided behaviour and perceptual harmony between the visible and tactile worlds. Although it has been suggested that seeing one's own body is crucial for adaptation, the underlying mechanisms are unclear. Here we show that a new representation of visuomotor mapping with respect to the hands emerges in a month during adaptation to reversed vision. The subjects become bi-perceptual, or able to use both new and old representations. In a visual task designed to assess the new hand representation, subjects identified visually presented hands as left or right by matching the picture to the representation of their own hands. Functional magnetic resonance imaging showed brain activity in the left posterior frontal cortex (Broca's area) that was unique to the new hand representations of both hands, together with activation in the intraparietal sulcus and prefrontal cortex. The emergence of the new hand representation coincided with the adaptation of perceived location of visible objects in space. These results suggest that the hand representation operates as a visuomotor transformation device that provides an arm-centred frame of reference for space perception. PMID- 11014193 TI - IRS-2 pathways integrate female reproduction and energy homeostasis. AB - Severe dietary restriction, catabolic states and even short-term caloric deprivation impair fertility in mammals. Likewise, obesity is associated with infertile conditions such as polycystic ovary syndrome. The reproductive status of lower organisms such as Caenorhabditis elegans is also modulated by availability of nutrients. Thus, fertility requires the integration of reproductive and metabolic signals. Here we show that deletion of insulin receptor substrate-2 (IRS-2), a component of the insulin/insulin-like growth factor-1 signalling cascade, causes female infertility. Mice lacking IRS-2 have small, anovulatory ovaries with reduced numbers of follicles. Plasma concentrations of luteinizing hormone, prolactin and sex steroids are low in these animals. Pituitaries are decreased in size and contain reduced numbers of gonadotrophs. Females lacking IRS-2 have increased food intake and obesity, despite elevated levels of leptin. Our findings indicate that insulin, together with leptin and other neuropeptides, may modulate hypothalamic control of appetite and reproductive endocrinology. Coupled with findings on the role of insulin-signalling pathways in the regulation of fertility, metabolism and longevity in C. elegans and Drosophila, we have identified an evolutionarily conserved mechanism in mammals that regulates both reproduction and energy homeostasis. PMID- 11014194 TI - Cell-fate conversion of lymphoid-committed progenitors by instructive actions of cytokines. AB - The primary role of cytokines in haemato-lymphopoiesis is thought to be the regulation of cell growth and survival. But the instructive action of cytokines in haematopoiesis has not been well addressed. Here we show that a clonogenic common lymphoid progenitor, a bone marrow-resident cell that gives rise exclusively to lymphocytes (T, B and natural killer cells), can be redirected to the myeloid lineage by stimulation through exogenously expressed interleukin (IL) 2 and GM-CSF (granulocyte/macrophage colony-stimulating factor) receptors. Analysis of mutants of the beta-chain of the IL-2 receptor revealed that the granulocyte- and monocyte-differentiation signals are triggered by different cytoplasmic domains, showing that the signalling pathway(s) responsible for these unique developmental outcomes are separable. Finally, we show that the endogenous myelomonocytic cytokine receptors for GM-CSF and macrophage colony-stimulating factor (M-CSF) are expressed at low to moderate levels on the more primitive haematopoietic stem cells, are absent on common lymphoid progenitors, and are upregulated after myeloid lineage induction by IL-2. We conclude that cytokine signalling can regulate cell-fate decisions and propose that a critical step in lymphoid commitment is downregulation of cytokine receptors that drive myeloid cell development. PMID- 11014195 TI - Tat-specific cytotoxic T lymphocytes select for SIV escape variants during resolution of primary viraemia. AB - Human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infections are characterized by early peaks of viraemia that decline as strong cellular immune responses develop. Although it has been shown that virus-specific CD8-positive cytotoxic T lymphocytes (CTLs) exert selective pressure during HIV and SIV infection, the data have been controversial. Here we show that Tat specific CD8-positive T-lymphocyte responses select for new viral escape variants during the acute phase of infection. We sequenced the entire virus immediately after the acute phase, and found that amino-acid replacements accumulated primarily in Tat CTL epitopes. This implies that Tat-specific CTLs may be significantly involved in controlling wild-type virus replication, and suggests that responses against viral proteins that are expressed early during the viral life cycle might be attractive targets for HIV vaccine development. PMID- 11014196 TI - Superoxide dismutase as a target for the selective killing of cancer cells. AB - Superoxide dismutases (SOD) are essential enzymes that eliminate superoxide radical (O2-) and thus protect cells from damage induced by free radicals. The active O2- production and low SOD activity in cancer cells may render the malignant cells highly dependent on SOD for survival and sensitive to inhibition of SOD. Here we report that certain oestrogen derivatives selectively kill human leukaemia cells but not normal lymphocytes. Using complementary DNA microarray and biochemical approaches, we identify SOD as a target of this drug action and show that chemical modifications at the 2-carbon (2-OH, 2-OCH3) of the derivatives are essential for SOD inhibition and for apoptosis induction. Inhibition of SOD causes accumulation of cellular O2- and leads to free-radical mediated damage to mitochondrial membranes, the release of cytochrome c from mitochondria and apoptosis of the cancer cells. Our results indicate that targeting SOD may be a promising approach to the selective killing of cancer cells, and that mechanism-based combinations of SOD inhibitors with free-radical producing agents may have clinical applications. PMID- 11014197 TI - A chemical switch for inhibitor-sensitive alleles of any protein kinase. AB - Protein kinases have proved to be largely resistant to the design of highly specific inhibitors, even with the aid of combinatorial chemistry. The lack of these reagents has complicated efforts to assign specific signalling roles to individual kinases. Here we describe a chemical genetic strategy for sensitizing protein kinases to cell-permeable molecules that do not inhibit wild-type kinases. From two inhibitor scaffolds, we have identified potent and selective inhibitors for sensitized kinases from five distinct subfamilies. Tyrosine and serine/threonine kinases are equally amenable to this approach. We have analysed a budding yeast strain carrying an inhibitor-sensitive form of the cyclin dependent kinase Cdc28 (CDK1) in place of the wild-type protein. Specific inhibition of Cdc28 in vivo caused a pre-mitotic cell-cycle arrest that is distinct from the G1 arrest typically observed in temperature-sensitive cdc28 mutants. The mutation that confers inhibitor-sensitivity is easily identifiable from primary sequence alignments. Thus, this approach can be used to systematically generate conditional alleles of protein kinases, allowing for rapid functional characterization of members of this important gene family. PMID- 11014198 TI - The protein Aly links pre-messenger-RNA splicing to nuclear export in metazoans. AB - In metazoans, most pre-messenger RNAs contain introns that are removed by splicing. The spliced mRNAs are then exported to the cytoplasm. Recent studies showed that splicing promotes efficient mRNA export, but the mechanism for coupling these two processes is not known. Here we show that Aly, the metazoan homologue of the yeast mRNA export factor Yralp (ref. 2), is recruited to messenger ribonucleoprotein (mRNP) complexes generated by splicing. In contrast, Aly does not associate with mRNPs assembled on identical mRNAs that already have no introns or with heterogenous nuclear RNP (hnRNP) complexes. Aly is recruited during spliceosome assembly, and then becomes tightly associated with the spliced mRNP. Aly shuttles between the nucleus and cytoplasm, and excess recombinant Aly increases both the rate and efficiency of mRNA export in vivo. Consistent with its splicing-dependent recruitment, Aly co-localizes with splicing factors in the nucleus. We conclude that splicing is required for efficient mRNA export as a result of coupling between the splicing and the mRNA export machineries. PMID- 11014199 TI - Chromodomains are protein-RNA interaction modules. AB - In Drosophila, compensation for the reduced dosage of genes located on the single male X chromosome involves doubling their expression in relation to their counterparts on female X chromosomes. Dosage compensation is an epigenetic process involving the specific acetylation of histone H4 at lysine 16 by the histone acetyltransferase MOF. Although MOF is expressed in both sexes, it only associates with the X chromosome in males. Its absence causes male-specific lethality. MOF is part of a chromosome-associated complex comprising male specific lethal (MSL) proteins and at least one non-coding roX RNA. How MOF is integrated into the dosage compensation complex is unknown. Here we show that association of MOF with the male X chromosome depends on its interaction with RNA. MOF specifically binds through its chromodomain to roX2 RNA in vivo. In vitro analyses of the MOF and MSL-3 chromodomains indicate that these chromodomains may function as RNA interaction modules. Their interaction with non coding RNA may target regulators to specific chromosomal sites. PMID- 11014200 TI - Image reconstructions of helical assemblies of the HIV-1 CA protein. AB - The type 1 human immunodeficiency virus (HIV-1) contains a conical capsid comprising approximately 1,500 CA protein subunits, which organizes the viral RNA genome for uncoating and replication in a new host cell. In vitro, CA spontaneously assembles into helical tubes and cones that resemble authentic viral capsids. Here we describe electron cryo-microscopy and image reconstructions of CA tubes from six different helical families. In spite of their polymorphism, all tubes are composed of hexameric rings of CA arranged with approximate local p6 lattice symmetry. Crystal structures of the two CA domains were 'docked' into the reconstructed density, which showed that the amino terminal domains form the hexameric rings and the carboxy-terminal dimerization domains connect each ring to six neighbours. We propose a molecular model for the HIV-1 capsid that follows the principles of a fullerene cone, in which the body of the cone is composed of curved hexagonal arrays of CA rings and the ends are closed by inclusion of 12 pentagonal 'defects'. PMID- 11014201 TI - Taiwan backs experience in quest for biotech success. PMID- 11014202 TI - Unfavourable economics put postdocs across Europe under strain. PMID- 11014203 TI - Postdocs reject academic research. PMID- 11014204 TI - The power of two--molecular differentiation of the vascular and bone actions of the natriuretic peptides. PMID- 11014205 TI - Hepatic growth hormone signaling in the late gestation fetal rat. AB - The role of GH in the developing fetus is poorly understood. Several studies have demonstrated a limited role for GH in late fetal life. In fact, few data are available regarding GH signal transduction in the late gestation fetus. We therefore focused on a comparison of hepatic GH signaling in near-term fetal rats [embryonic day 19 (E19)] and adult rats using a combination of in vitro studies employing hepatocytes in primary culture and in vivo studies. We found that GH receptor (GHr) binding was comparable in fetal liver and adult liver. The long isoform of the GHr underwent tyrosine phosphorylation in response to GH stimulation of E19 fetal hepatocytes in a manner similar to that seen in cultured adult hepatocytes. Furthermore, downstream signaling via the Janus kinase-2 tyrosine kinase, STAT1 (signal transducer and activator of transcription), and STAT5 was also intact in both, as demonstrated by the tyrosine phosphorylation of these signaling proteins. To confirm the relevance of these findings to the in vivo situation, GH was directly administered by ip injection to E 19 fetal and adult rats. In both cases, tyrosine phosphorylation of STAT5 was markedly and rapidly induced. Finally, transfection of E19 fetal hepatocytes with GH responsive reporter elements [Spi2.1(-275/+85)-CAT and 8xGHRE-TKCAT] demonstrated intact transcriptional regulation. Our data indicate that GHr abundance and activity as well as downstream GH signaling are similar in the late gestation fetal rat and in the adult and that these mechanisms appear capable of supporting physiological GH functions in the developing liver. PMID- 11014206 TI - Conformational changes and coactivator recruitment by novel ligands for estrogen receptor-alpha and estrogen receptor-beta: correlations with biological character and distinct differences among SRC coactivator family members. AB - Ligands for the estrogen receptor (ER) that have the capacity to selectively bind to or activate the ER subtypes ERalpha or ERbeta would be useful in elucidating the biology of these two receptors and might assist in the development of estrogen pharmaceuticals with improved tissue selectivity. In this study, we examine three compounds of novel structure that act as ER subtype-selective ligands. These are a propyl pyrazole triol (PPT), which is a potent agonist on ERalpha but is inactive on ERbeta, and a pair of substituted tetrahydrochrysenes (THC), one enantiomer of which (S,S-THC) is an agonist on both ERalpha and ERbeta, the other (R,R-THC) being an agonist on ERalpha but an antagonist on ERbeta. To investigate the molecular mechanisms underlying the ER subtype selective actions of these compounds, we have determined the conformational changes induced in ERalpha and ERbeta by these ligands using protease digestion sensitivity, and we have tested the ability of these ligands to promote the recruitment of representatives of the three SRC/p160 coactivator protein family members (SRC-1, GRIP-1, ACTR, respectively) to ERalpha and ERbeta using yeast two hybrid and glutathione-S-transferase (GST) pull-down assays. We find that the ligand-ER protease digestion pattern is distinctly different for stimulatory and inhibitory ligands, and that this assay, as well as coactivator recruitment, are excellent indicators of their agonist/antagonist character. Interestingly however, compared with estradiol, the novel agonist ligands show some quantitative differences in their ability to recruit SRC-1, -2, and -3. This implies that while generally similar to estradiol, these ligands induce ER conformations that differ somewhat from that induced by estradiol, differences that are illustrative of the nature of their biological character. The application of methods to characterize the conformations induced in ER subtypes by novel ligands, as done in this study, enables a greater understanding of how ligand-receptor conformations relate to estrogen agonist or antagonist behavior. PMID- 11014207 TI - Double-stranded ribonucleic acid decreases C6 rat glioma cell numbers: effects on insulin-like growth factor I gene expression and action. AB - Poly(IC), a synthetic double-stranded RNA copolymer of inosinic and cytidilic acids, decreases the growth of normal and tumorigenic cells. We tested the hypothesis that Poly(IC) decreases C6 glioma cell growth by disrupting an autocrine insulin-like growth factor I (IGF-I) growth loop. Addition of Poly(IC) decreased C6 cell number in confluent and sparse cultures in a dose-dependent manner. Addition of exogenous IGF-I partially compensated for the decrease in cell number caused by Poly(IC) in confluent and subconfluent cultures of C6 cells, suggesting that one mechanism of Poly(IC) action is through down regulation of IGF-I gene expression and/or action. Treatment of confluent C6 cells with 10 and 200 microg/ml Poly(IC) for 24 h decreased IGF-I messenger RNA (mRNA) levels to 50% and 25% of the control value, respectively. Treatment of C6 cells with 200 microg/ml Poly(IC) for 24 h reduced IGF-I receptor mRNA levels to 50% of the control level. IGF-binding protein-1 (IGFBP-1), -2, and -6 mRNAs were not expressed in the C6 cells used in this study. Treatment of C6 cells with 200 microg/ml Poly(IC) for 24 h reduced IGFBP-4 mRNA and IGFBP-5 mRNA levels to 26% and 29% of the control level, respectively. There was no significant change in IGFBP-3, insulin receptor, or actin mRNA levels with Poly(IC) treatment. Treatment of confluent C6 cells with 200 microg/ml Poly(IC) for 24 h decreased levels of immunoreactive IGF-I in conditioned medium (CM) to 55% of the control value, decreased IGF-I receptor beta-subunit levels to 28% of the control value, and decreased levels of IGFBP-3, IGFBP-4, and IGFBP-5 protein in CM to 45%, 50%, and 30% of the control values, respectively. There was no significant change in actin and tubulin protein levels with Poly(IC) treatment. These results suggest that IGF-I gene expression is down-regulated by Poly(IC) treatment and that IGF-I bioavailability and action in C6 cells are also altered due to decreases in IGF-I receptor and binding protein levels. PMID- 11014208 TI - Modulation of pituitary somatostatin receptor subtype (sst1-5) messenger ribonucleic acid levels by changes in the growth hormone axis. AB - The role of individual components of the hypothalamic-pituitary-GH axis in the modulation of pituitary somatostatin (SRIF) receptor subtype (sst1-5) synthesis was assessed using multiplex RT-PCR to measure receptor messenger RNA (mRNA) levels in normal rats and spontaneous dwarf rats (SDRs). In SDRs, a strain with no immunodetectable GH, pituitary sst1 and sst2 mRNA levels were elevated, sst5 mRNA levels were reduced, and sst3 and sst4 mRNA levels did not significantly differ from those in normal controls. Treatment of SDRs with GH (72 h), but not insulin-like growth factor I, significantly decreased sst2 mRNA levels and increased sst4 and sst5 mRNA levels above vehicle-treated control levels. To test whether more rapid changes in circulating GH levels could alter SRIF receptor subtype expression, normal rats were infused (iv) with GH-releasing hormone (GHRH) for 4 h in the presence or absence of SRIF antiserum. GHRH infusion increased pituitary sst1 and sst2 and decreased sst5, but had no effect on sst3 and sst4 mRNA levels. Immunoneutralization of SRIF, which produced a rise in circulating GH levels, did not alter basal or GHRH-mediated SRIF receptor subtype expression. These observations indicate that acute suppression of SRIF tone does not regulate pituitary SRIF receptor subtype mRNA levels in vivo. The possibility that elevated circulating GH concentrations induced by GHRH infusion were responsible for the observed changes in SRIF receptor subtype mRNA levels was examined by infusing SDRs with GHRH for 4 h. GHRH did not increase sst1 mRNA levels in SDRs above their already elevated value. However, GHRH infusion produced an increase in sst2 and a decrease in sst5 mRNA levels similar to those observed in normal rats, indicating that the acute effects of GHRH on SRIF receptor subtype expression are independent of circulating GH levels. Primary rat pituitary cell cultures were incubated with GHRH (10 nM) or forskolin (10 microM) for 4 h to determine whether GHRH could directly mediate SRIF receptor subtype mRNA. GHRH treatment increased sst1 and sst2 mRNA levels and decreased sst5 mRNA levels, but had no effect on sst3 and sst4, similar to the results in vivo. The effect of forskolin mimicked that of GHRH on sst1, sst2, and sst5 mRNA, suggesting that GHRH acts through cAMP to directly mediate gene transcription or mRNA stability of these SRIF receptor subtypes. In addition, forskolin reduced sst3 and sst4 expression. These results strongly suggest that rat pituitary sst1, sst2, and sst5 mRNA levels are regulated both in vivo and in vitro by GHRH. The stimulatory action of GHRH on sst1 and sst2 and the inhibitory action on sst5 indicate that these receptor subtypes have independent and unique roles in the modulation of pituitary GH release. PMID- 11014209 TI - Prolactin (PRL) receptor gene expression in mouse adipose tissue: increases during lactation and in PRL-transgenic mice. AB - There are indications that PRL may exert important metabolic actions on adipose tissue in different species. However, with the exception of birds, the receptor has not been identified in white adipose tissue. The present study was designed to examine the possible expression and regulation of the PRL receptor (PRLR) in mouse adipose tissue. The long PRLR messenger RNA (mRNA) splice form (L-PRLR) and two short splice forms (S2- and S3-PRLR) were detected in mouse adipose tissue by RT-PCR. Furthermore, L-PRLR mRNA was detected by ribonuclease protection assay. Immunoreactive PRLR with a relative molecular mass of 95,000 was revealed by immunoblotting. Furthermore, L-PRLR mRNA expression was demonstrated in primary isolated adipocytes. In mouse adipose tissue, the level of L-PRLR mRNA expression increased 2.3-fold during lactation compared with those in virgin and pregnant mice. In contrast, in the liver the expression of L-PRLR increased 3.4-fold during pregnancy compared with those in virgin and lactating mice. When comparing the levels of L-PRLR expression in virgin female and male mice, no difference was detected in adipose tissue. However, in virgin female liver the expression was 4.5-fold higher than that in male liver. As PRL up-regulates its own receptor in some tissues, we analyzed L-PRLR expression in PRL-transgenic female and male mice. In PRL-transgenic mice L-PRLR expression was significantly increased in both adipose tissue (1.4-fold in females and 2.4-fold in males) and liver (1.9 fold in females and 2.7-fold in males) compared with that in control mice. Furthermore, in female PRL-transgenic mice retroperitoneal adipose tissue was decreased in weight compared with that in control mice. However, no difference was detected when comparing the masses of parametrial adipose tissue. Our results suggest a direct role for PRL, mediated by PRLR, in modulating physiological events in adipose tissue. PMID- 11014210 TI - Evidence that cleavage of the thyrotropin receptor involves a "molecular ruler" mechanism: deletion of amino acid residues 305-320 causes a spatial shift in cleavage site 1 independent of amino acid motif. AB - Some TSH receptors (TSHR) on the cell surface cleave into A and B subunits. Cleavage at upstream Site 1 is followed by the proteolytic excision of an intervening C peptide region terminating at a downstream Site 2. Although present evidence suggests that Site 1 lies between amino acid residues 303 and 317, the mechanism and exact amino acid(s) involved in cleavage are unknown. Previous amino acid substitutions at Site 1 failed to abrogate cleavage. We, therefore, performed deletion mutations within this region. Cleavage of cell surface TSHR, detected by 125I-TSH cross-linking to intact cells, was not prevented by deletion of four individual segments within the Site 1 cleavage region (delta305-308, delta309-312, delta313-316, delta317-320). However, deletion of the entire region (delta305-320) reduced the extent of cleavage and shifted the cleavage site upstream of the glycan at amino acid residue N302. Elimination of this glycan (N302Q substitution) reversed the effect of deleting amino acid residues 305-320 on TSHR cleavage, suggesting that reduced cleavage at the new, upstream cleavage site was caused by steric hindrance by the glycan at N302. In summary, deletion, as opposed to mutagenesis, of the TSHR cleavage Site 1 region produces a spatial shift in TSHR cleavage Site 1 from downstream to upstream of the glycan at N302. These observations provide strong evidence that TSHR cleavage at this site does not occur at a particular amino acid motif and suggests that cleavage involves a "molecular ruler" mechanism involving cleavage at a fixed distance from a protease attachment site. PMID- 11014211 TI - Proliferation of mouse mammary epithelial cells in vitro: interactions among epidermal growth factor, insulin-like growth factor I, ovarian hormones, and extracellular matrix proteins. AB - The purpose of the present study was to investigate the role of extracellular matrix proteins (ECMs; collagens I and IV, fibronectin, and laminin) in modulating proliferative responses of normal mammary epithelial cells in serum free culture to epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I). As EGF and IGF-I can alter steroid responses, the interactions among growth factors, estrogen, and R5020 were also investigated. We report the novel finding that all ECMs tested, but not a nonspecific attachment factor, poly-L lysine (PL), promoted a highly synergistic proliferative response to EGF plus IGF I. EGF receptors were significantly increased with culture time on all ECMs, but not on PL. IGF receptor expression was significantly 2- to 4-fold higher on all ECMs compared with PL. EGF decreased IGF-binding protein-2 (IGFBP-2) and IGFBP-3 by more than 50% in the presence of IGF-I on PL or collagen I. These results indicate that ECM-specific IGF-I/EGF synergism occurs in response to ECM up regulation of growth factor receptors and EGF down-regulation of inhibitory IGFBPs. Growth factors did not synergize with estrogen and/or R5020. Instead, estrogen plus R5020 decreased EGF-plus IGF-I-induced proliferation in an ECM dependent manner. These studies demonstrate that proliferation of normal mammary epithelial cells involves complex interactions among steroids, growth factors, binding proteins, and ECMs. PMID- 11014212 TI - Transcriptional regulation of human 11beta-hydroxylase (hCYP11B1). AB - Steroid 11beta-hydroxylase is a mitochondrial enzyme that catalyzes the conversion of deoxycortisol to cortisol. The gene encoding human 11beta hydroxylase (hCYP11B1) is expressed in the adrenal cortex under the control of circulating levels of ACTH. The current study was undertaken to define the cis regulatory elements and transacting factors that regulate hCYP11B1 transcription. The hCYP11B1 5'-flanking DNA was studied using transient transfection of luciferase reporter constructs in NCI-H295R human adrenocortical cells. A cAMP analogue ((Bu)2cAMP) increased expression of a construct containing -1102 bp of hCYP11B1 5'-flanking DNA (pB1-1102). An element at position -71/-64 (TGACGTGA, previously termed Ad1) resembling a consensus cAMP response element (CRE) was required for maximal induction by cAMP. The Ad1 element bound several transcriptional factors in electrophoretic mobility shift assays, including CRE binding protein, activating transcription factor-1 (ATF-1), and ATF-2, but only the ATF-2 complex migrated similarly to a complex seen using H295R nuclear extract. In addition, Western analysis of H295R and adrenal lysates demonstrated expression of high levels of ATF-2 and ATF-1. CRE-binding protein levels varied among the strains of H295R cells tested. Transcription of CYP11B1 also appeared to be regulated by steroidogenic factor-1 (SF-1). Luciferase reporter gene activity was increased after cotransfection with expression vectors containing SF 1. An element in hCYP11B1 at positions 242/-234 (CCAAGGCTC), previously termed Ad4, was required for maximal induction by SF-1 and was found to bind SF-1 in electrophoretic mobility shift assays. The key role for SF-1 in hCYP11B1 transcription is in contrast to its lack of an effect on expression of the hCYP11B2 (aldosterone synthase) isozyme. The differential effects of SF-1 on transcription of hCYP11B1 and hCYP11B2 may be one of the mechanisms controlling differential expression of these isozymes within the zonae fasciculata and glomerulosa of the human adrenal cortex. PMID- 11014213 TI - Effects of arsenite on estrogen receptor-alpha expression and activity in MCF-7 breast cancer cells. AB - To determine whether arsenite has estrogen-like activities, the effects of this compound on estrogen receptor-alpha (ERalpha) and other estrogen-regulated genes were measured in the human breast cancer cell line MCF-7. Treatment of cells with 1 microM arsenite resulted in a 60% decrease in the amount of ERalpha and in a parallel decrease of 40% in ERalpha messenger RNA. Progesterone receptor concentration increased 22-fold after arsenite treatment. pS2 messenger RNA also increased 2. 1-fold after treatment. The induction of progesterone receptor and pS2 was blocked by the antiestrogen ICI-182,780. In transient cotransfection experiments of wild-type ERalpha and an estrogen response element-reporter construct, arsenite stimulated chloramphenicol acetyltransferase (CAT) activity. In growth assays, arsenite significantly stimulated the proliferation of MCF-7 cells compared with cells grown in estrogen-depleted medium. Addition of an antiestrogen blocked growth stimulation by arsenite. In binding assays, arsenite blocked the binding of estradiol to ERalpha (Ki = 5 +/- 0.5 nM; n = 3), suggesting that the compound interacts with the hormone-binding domain of the receptor. To determine whether interaction of arsenite with the hormone-binding domain results in receptor activation, COS-1 cells were transiently cotransfected with the chimeric receptors GAL-ER, which contains the hormone-binding domain of ERalpha and the DNA-binding domain of the transcription factor GAL4, and a GAL4 responsive CAT reporter gene. Treatment of cells with estradiol or arsenite resulted in a 4-fold increase in CAT activity. The effects of arsenite on the chimeric receptor were blocked by the antiestrogen, suggesting that arsenite activates ERalpha through an interaction with the hormone-binding domain of the receptor. Transfection assays with ERalpha mutants identified C381, C447, H524, and N532 as interaction sites of arsenite with the hormone-binding domain. PMID- 11014214 TI - Cytological characterization of a pituitary folliculo-stellate-like cell line, Tpit/F1, with special reference to adenosine triphosphate-mediated neuronal nitric oxide synthase expression and nitric oxide secretion. AB - An immortal nonhormone-producing cell line with a characteristic star-shaped morphology, named Tpit/F1, was derived from an anterior pituitary gland of a temperature-sensitive large T antigen transgenic mouse. To characterize Tpit/F1 cells, we performed cytological studies, which revealed that Tpit/F1 cells express the messenger RNAs of neruonal nitric oxide (NO) synthase, S-100 protein, basic fibroblast growth factor, and pituitary-restricted transcription factor. The Tpit/F1 cells response to pituitary adenylate cyclase-activating peptide comprised the stimulated secretion of interleukin-6. Furthermore, glucocorticoids stimulate glutamine synthase production by Tpit/F1 cells. Considering these cytological characteristics together with their morphology, we deduced that Tpit/F1 cells are derived from pituitary folliculo-stellate (FS) cells. Our cytophysiological analyses of Tpit/F1 cells revealed that intracellular Ca2+ increased dose dependently on ATP administration (0-100 microM), and that this effect did not require the presence of extracellular Ca2+ and was not abolished by treatment with gadolinium, a Ca2+ channel blocker. The ATP-induced increase in intracellular Ca2+ ([Ca2+]i) was completely abolished by treatment with the Ca2+ adenosine triphosphatase (Ca2+-ATPase) inhibitor thapsigargin, which suggests that ATP increases [Ca2+]i by mobilizing internally stored Ca2+ followed by an influx of Ca2+. Moreover, UTP was equipotent with ATP in causing the [Ca2+]i increase in Tpit/F1 cells. Also, the Ca2+ response was prevented by the phospholipase C inhibitor, U-73122, but not by its inactive analog, U-73343. From these results we therefore concluded that ATP acts on Tpit/F1 cells via P2Y2 purinoceptors. Interestingly, both neuronal nitric oxide synthase messenger RNA and NO secretion were increased by ATP administration (10 and 100 microM). These results suggest the biological significance of the topological colocalization of FS cells and endocrine cells. Namely, ATP is cosecreted with hormones from endocrine cells and stimulates NO production by FS cells, and the released NO may regulate neighboring endocrine cell and blood vessels. PMID- 11014215 TI - Transcriptional down-regulation of human gonadotropin-releasing hormone (GnRH) receptor gene by GnRH: role of protein kinase C and activating protein 1. AB - Clinical applications of GnRH agonists (GnRHa) are based primarily on the decrease in gonadotropin release after down-regulation of the GnRH receptor (GnRHR) by continuous GnRHa administration. However, the molecular mechanisms underlying the transcriptional regulation of the human GnRHR gene after prolonged GnRH treatment remain poorly understood. In the present study GnRHa-mediated regulation of human GnRHR gene transcription was studied by transiently transfecting the mouse gonadotrope-derived (alphaT3-1) cells with a 2297-bp human GnRHR promoter-luciferase construct (p2300-LucF). A dose- and time-dependent decrease in human GnRHR promoter activity was observed after GnRHa treatment. An average 71% decrease in promoter activity was observed after 24-h treatment with 0.1 microM GnRHa, which was blocked by cotreatment of the GnRH antagonist, antide. This effect was mimicked by phorbol 12-myristate 13-acetate (TPA) administration. In addition, the GnRHa- and TPA-mediated decrease in the human GnRHR promoter activity was reversed by a specific protein kinase C (PKC) inhibitor, GF109203X, or depletion of PKC by TPA pretreatment. These findings indicate that the activation of the PKC pathway is important in regulating the human GnRHR gene expression. By progressive 5'-deletion studies, we have identified a 248-bp DNA fragment (-1018 to -771, relative to the translation start site) at the 5'-flanking region of the human GnRHR gene that is responsible for the GnRHa-mediated down-regulation of human GnRHR promoter activity. Analysis of this sequence reveals the existence of two putative activating protein-1 (AP 1) sites with 87% homology to the consensus sequence (5'-TGA(G/C)T(C/A)A-3'), located at -1000 to -994 (5'-TTAGACA-3', in complementary orientation) and -943 to 937 (5'-TGAATAA-3'). Using competitive gel mobility shift assays, AP-1 binding was observed within this 248-bp region. Site-directed mutation of the putative AP 1-binding site located at -1000 to -994 abolished the GnRHa-induced inhibition. Further competitive GMSA and supershift experiments confirmed the identity of AP 1 binding in this region. By the use of Western blot analysis, a significant increase in c-Jun (100%; P < 0.05) and c-Fos (50%; P < 0.05) protein levels was observed after GnRHa treatment in alphaT3-1 cells. In addition, our data suggested that a change in AP-1 composition, particularly c-Fos, was important in mediating GnRHa-induced inhibition of human GnRHR gene expression. We conclude that activation of the PKC pathway by GnRH is important in controlling human GnRHR gene expression. In addition, the putative AP-1-binding site located at 1000 to -994 of the human GnRHR5'-flanking region has been functionally identified to be involved in mediating this down-regulatory effect. PMID- 11014216 TI - Nicotine up-regulates expression of orexin and its receptors in rat brain. AB - Orexins are two recently discovered neuropeptides that can stimulate food intake. As the chronic use of tobacco typically leads to a reduction in body weight, it is of interest to determine whether nicotine, the major biologically active tobacco ingredient, has an effect on orexin metabolism in the brain. Using a semiquantitative RT-PCR technique, the levels of messenger RNA (mRNA) for prepro orexin, orexin A (OX1-R) and orexin B (OX2-R) receptors were 20-50% higher in rats receiving nicotine for 14 days at the level of 2-4 mg/kg day compared with rats receiving saline solvent alone. In animals treated with nicotine at 4 mg/kg x day, the expression levels of mRNA for prepro-orexin, OX1-R, and OX2-R were significantly higher compared with those in either the free-feeding control or pair-fed saline control rats. RIA data indicated that both orexin A and orexin B peptide levels were significantly elevated (45-54%; P < 0.01) in the dorsomedial nucleus (DMH) of the nicotine-treated rats compared with either solvent-only or pair-fed controls. Additionally, orexin B was significantly elevated (83%; P < 0.01), over levels in both types of the control animals, in the paraventricular nucleus (PVN) region. In summary, we demonstrated that an inverse association between nicotine and food intake as well as body weight held with doses comparable to those consumed by average human smokers. Moreover, our data indicated that chronic exposure to nicotine can induce a long-term increase in the expression levels of prepro-orexin and their receptor mRNA in the rat hypothalamus and in the levels of orexin A in the DMH and orexin B in the DMH and PVN among the six hypothalamic regions that we examined. PMID- 11014217 TI - Diazoxide restores beta3-adrenergic receptor function in diet-induced obesity and diabetes. AB - We previously demonstrated that the expression and function of the adipocyte specific beta3-adrenergic receptor (beta3AR) are significantly depressed in single gene and diet-induced rodent models of obesity. Furthermore, these models are relatively unresponsive to the anti-obesity effects of beta3AR agonists. Because all of these models are hyperinsulinemic, we hypothesized that hyperinsulinemia could be responsible for this abnormality in beta3AR function. The goal of this study was to determine whether lowering insulin with the K-ATP channel agonist, diazoxide (Dz) would reverse the depressed expression and function of the beta3AR found in a model of diet-induced diabetes and obesity in C57BL/6J (B6) mice. B6 male mice were placed on either high fat (HF) or low fat experimental diets. After 4 weeks, HF-fed mice were assigned to a group: HF or HF containing disodium (R,R)-5- [2-( [2-(3-chlorophenyl)-2-hydroxyethyl] amino]propyl-1,3-benzodioxole-2,2-di carboxylate (CL; 0.001%, wt/wt), Dz (0.32%, wt/wt), or their combination (CLDz). Dz animals exhibited significantly reduced plasma insulin levels as well as increased 3pAR expression and agonist-stimulated adenylyl cyclase activity in adipocytes. CLDz was more effective in reducing percent body fat, lowering nonesterified fatty acids, improving glucose tolerance, and reducing feed efficiency than either treatment alone. PMID- 11014218 TI - Expression of enzymes synthesizing (aldehyde dehydrogenase 1 and reinaldehyde dehydrogenase 2) and metabolizaing (Cyp26) retinoic acid in the mouse female reproductive system. AB - Vitamin A is required for female reproduction. Rodent uterine cells are able to synthesize retinoic acid (RA), the active vitamin A derivative, and express RA receptors. Here, we report that two RA-synthesizing enzymes [aldehyde dehydrogenase 1 (Aldh1) and retinaldehyde dehydrogenase 2 (Raldh2)] and a cytochrome P450 (Cyp26) that metabolizes vitamin A and RA into more polar metabolites exhibit dynamic expression patterns in the mouse uterus, both during the ovarian cycle and during early pregnancy. Aldh1 expression is up-regulated during diestrus and proestrus in the uterine glands, whereas Raldh2 is highly induced in the endometrial stroma in metestrus. Cyp26 expression, which is not detectable during the normal ovarian cycle, is strongly induced in the uterine luminal epithelium, 24 h after human CG hormonal administration. Raldh2 stromal expression also strongly responds to gonadotropin (PMSG and human CG) induction. Furthermore, Raldh2 expression can be hormonally induced in stromal cells of the vagina and cervix. All three enzymes exhibit differential expression profiles during early pregnancy. Aldh1 glandular expression is sharply induced at 2.5 gestational days, whereas Raldh2 stromal expression increases more steadily until the implantation phase. Cyp26 epithelial expression is strongly induced between 3.5-4.5 gestational days, i.e. when the developing blastocysts colonize the uterine lumen. These data suggest a need for precise regulation of RA synthesis and/or metabolism, in both cycling and pregnant uterus. PMID- 11014219 TI - Antiinflammatory effects of estrogen on microglial activation. AB - In the present study the effects of 17beta-estradiol on microglial activation are described. Estrogen replacement therapy has been associated with decreased severity of age-related neurodegenerative diseases such as Alzheimer's disease, and estrogens have potent immunosuppressive properties outside of the brain. To determine the role that microglial cells might play in estrogen-mediated neuroprotection, primary rat microglia and N9 microglial cell lines were treated with increasing doses of 17beta-estradiol before or during immunostimulation by lipopolysaccharide, phorbol ester, or interferon-gamma. Pretreatment with 17beta estradiol, but not 17alpha-estradiol or progesterone, dose dependently attenuated microglial superoxide release and phagocytic activity. Additionally, 17beta estradiol attenuated increases in inducible nitric oxide synthase protein expression, but did not alter nuclear factor-KB activation. The antiinflammatory effects of 17beta-estradiol were blocked by the antiestrogen ICI 182,780. Additionally, 17beta-estradiol induced rapid phosphorylation of the p42/p44 mitogen-activated protein kinase (MAP kinase), and the MAP kinase inhibitor PD 98059 blocked the antiinflammatory effects of 17beta-estradiol. Overall, these results suggest that estrogen receptor-dependent activation of MAP kinase is involved in estrogen-mediated antiinflammatory pathways in microglial cells. These results describe a novel mechanism by which estrogen may attenuate the progression of neurodegenerative disease and suggest new pathways for therapeutic intervention in clinical settings. PMID- 11014220 TI - Resveratrol acts as a mixed agonist/antagonist for estrogen receptors alpha and beta. AB - Epidemiological evidence indicates that phytoestrogens inhibit cancer formation and growth, reduce cholesterol levels, and show benefits in treating osteoporosis. At least some of these activities are mediated through the interaction of phytoestrogens with estrogen receptors alpha and beta (ERalpha and ERbeta). Resveratrol, trans-3,5,4'-trihydroxystilbene, is a phytoestrogen in grapes that is present in red wine. Resveratrol was shown to bind ER in cytosolic extracts from MCF-7 and rat uteri. However, the contribution of ERalpha vs. ERbeta in this binding is unknown. Here we report that resveratrol binds ERbeta and ERalpha with comparable affinity, but with 7,000-fold lower affinity than estradiol (E2). Thus, resveratrol differs from other phytoestrogens that bind ERbeta with higher affinity than ERalpha. Resveratrol acts as an estrogen agonist and stimulates ERE-driven reporter gene activity in CHO-K1 cells expressing either ERalpha or ERbeta. The estrogen agonist activity of resveratrol depends on the ERE sequence and the type of ER. Resveratrol-liganded ERbeta has higher transcriptional activity than E2-liganded ERbeta at a single palindromic ERE. This indicates that those tissues that uniquely express ERbeta or that express higher levels of ERbeta than ERalpha may be more sensitive to resveratrol's estrogen agonist activity. For the natural, imperfect EREs from the human c-fos, pS2, and progesterone receptor (PR) genes, resveratrol shows activity comparable to that induced by E2. We report that resveratrol exhibits E2 antagonist activity for ERalpha with select EREs. In contrast, resveratrol shows no E2 antagonist activity with ERbeta. These data indicate that resveratrol differentially affects the transcriptional activity of ERalpha and ERbeta in an ERE sequence-dependent manner. PMID- 11014221 TI - Neuroendocrine cell type-specific and inducible expression of the chromogranin B gene: crucial role of the proximal promoter. AB - Chromogranin B, a soluble acidic secretory protein, is widely distributed in neuroendocrine and neuronal cells, although not in other cell types. To identify the elements governing such widespread, yet selective, expression of the gene, we characterized the isolated mouse chromogranin B promoter. 5'-Promoter deletions localized neuroendocrine cell type-specific expression to the proximal chromogranin B promoter (from -216 to -91 bp); this region contains an E box (at [-206 bp]CACCTG[-201 bp]), four G/C-rich regions (at [-196 bp]CCCCGC[-191 bp], [ 134 bp]CCGCCCGC[-127 bp], [-125 bp]GGCGCCGCC[-117 bp], and [-115 bp]CGGGGC[-110 bp]), and a cAMP response element (CRE; at [-102 bp]TGACGTCA[-95 bp]). A 60-bp core promoter region, defined by an internal deletion from - 134 to -74 bp upstream of the cap site and spanning the CRE and three G/C-rich regions, directed tissue-specific expression of the gene. The CRE motif directed cell type specific expression of the chromogranin B gene in neurons, whereas three of the G/C-rich regions played a crucial role in neuroendocrine cells. Both the endogenous chromogranin B gene and the transfected chromogranin B promoter were induced by preganglionic secretory stimuli (pituitary adenylyl cyclase-activating polypeptide, vasoactive intestinal peptide, or a nicotinic cholinergic agonist), establishing stimulus-transcription coupling for this promoter. The adenylyl cyclase activator forskolin, nerve growth factor, and retinoic acid also activated the chromogranin B gene. Secretagogue-inducible expression of chromogranin B also mapped onto the proximal promoter; inducible expression was entirely lost upon internal deletion of the 60-bp core (from 134 to -74 bp). We conclude that CRE and G/C-rich domains are crucial determinants of both cell type specific and secretagogue-inducible expression of the chromogranin B gene. PMID- 11014222 TI - Transforming growth factor-beta2 mediates mesenchymal-epithelial interactions of testicular somatic cells. AB - Transforming growth factor-beta2 (TGFbeta2) is an important mediator of growth and differentiation. We here describe for the first time the complete sequence of the TGFbeta2 complementary DNA derived from peritubular myoid cells of the rat testis. The size of the rat TGFbeta2 complementary DNA was 1245 bp, and the deduced protein sequence contained 414 amino acids. Sequence comparison with the human and mouse amino acid sequences demonstrated 96.4% and 97.9% sequence identities, respectively. To elucidate the functional role of TGFbeta2 in testicular somatic cells, we studied its secretion in vitro in monocultures and cocultures of mesenchymal peritubular and epithelial Sertoli cells. The highest amounts of TGFbeta2 protein were secreted in the cocultures and by peritubular cells, whereas Sertoli cells secreted only minor amounts. Stimulation experiments with FSH revealed a reduced secretion of TGFbeta2 in cocultures, probably mediated by a paracrine interaction of the FSH-responsive Sertoli cells. In contrast, TGFbeta2 secretion by peritubular cells was increased after stimulation with glucocorticoids and after addition of recombinant TGFbeta2, indicating an autoregulation of TGFbeta2. Furthermore, application of recombinant TGFbeta2 to cocultures resulted in an enhanced aggregation and cell clustering of Sertoli cells, pointing to an important role of TGFbeta2 in the paracrine interaction of peritubular and Sertoli cells of the developing rat testis. PMID- 11014223 TI - Potentiation of growth hormone-induced liver suppressors of cytokine signaling messenger ribonucleic acid by cytokines. AB - Endotoxin and proinflammatory cytokines such as interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha) induce a state of GH resistance. A new family of suppressors of cytokine signaling (SOCS), induced by cytokines activating the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway, has been recently identified as a negative feedback loop of intracellular signaling. Overexpression of some SOCS (SOCS-3, CIS, and SOCS-2) has been reported to inhibit the JAK-STAT pathway stimulated by GH. To assess the possible role of these three SOCS proteins in the GH resistance induced by endotoxin and cytokines, we investigated the regulation of their gene expression by endotoxin and GH in rat liver and by proinflammatory cytokines and GH in primary culture hepatocytes. Both GH and lipopolysaccharide induced the three SOCS messenger RNAs (mRNAs) in vivo. In vitro, GH also increased the liver mRNAs encoding SOCS-2, SOCS-3, and CIS. Although IL-1/beta and TNFalpha alone induced only weakly the expression of SOCS-3 and CIS, these cytokines strongly potentiated the induction of these two SOCS by GH. In contrast, IL-6 alone markedly induced SOCS-3 mRNA, but did not potentiate the GH action on SOCS-3 and CIS mRNAs. The GH induction of SOCS-2 was not potentiated by any of these cytokines. Considering the ability of these SOCS to inhibit the JAK-STAT pathway induced by GH, these results suggest that the overexpression of SOCS-3 and CIS mRNAs induced by IL-1beta and TNFalpha or by endotoxin in vivo may play a role in the GH resistance induced by sepsis. PMID- 11014224 TI - Induction of calcitonin and calcitonin receptor expression in rat mammary tissue during pregnancy. AB - Human breast milk samples at early time points after parturition contain high levels of calcitonin (CT) in both normal and thyroidectomized mothers, suggesting that mammary tissue produces CT. Using blot hybridization and reverse transcriptase polymerase chain (RT-PCR) analysis of rat mammary RNA we found that CT messenger RNA is induced at midpregnancy (day 12), remains elevated through late pregnancy (day 19) but then decreases before the day of birth. RIA of mammary CT revealed that levels increase from 0.3 ng/g tissue in nonpregnant animals to peak at 1.6 ng/g on day 19 and then decline after that, paralleling messenger RNA expression. Dilution profiles for extracted mammary CT showed close parallelism with monomeric rat CT. Plasma samples from thyroparathyroidectomized rats contained 10-20 pg/ml CT that did not increase during pregnancy, suggesting that mammary CT is not released into plasma but functions locally. Consistent with this, RT-PCR detected that the CT receptor C1a isoform is expressed in rat mammary tissues during both pregnancy and lactation. This is the first report that mammary tissue expresses both CT and the CT receptor during pregnancy, suggesting that CT may have a paracrine regulatory role in the mammary gland. PMID- 11014225 TI - Glucagon-like peptide-1, but not glucose-dependent insulinotropic peptide, regulates fasting glycemia and nonenteral glucose clearance in mice. AB - Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) potentiate glucose-stimulated insulin secretion after enteral nutrient ingestion. We compared the relative incretin and nonincretin actions of GLP-1 and GIP in +/+ and GLP-1R-/- mice using exendin(9-39) and immunopurified anti-GIP receptor antisera (GIPR Ab) to antagonize GLP-1 and GIP action, respectively. Both antagonists produced a significant increase in glycemic excursion after oral glucose loading of +/+ mice (P < 0.05 for antagonists us. controls). Exendin(9 39) also increased blood glucose and decreased glucose-stimulated insulin in +/+ mice after ip glucose loading [0.58 +/- 0.02 vs. 0.47 +/- 0.02 ng/ml in saline- vs. exendin(9-39)-treated mice, respectively, P < 0.05]. In contrast, GIPR Ab had no effect on glucose excursion or insulin secretion, after ip glucose challenge, in +/+ or GLP-1R-/- mice. Repeated administration of exendin(9-39) significantly increased blood glucose and reduced circulating insulin levels but had no effect on levels of pancreatic insulin or insulin messenger RNA transcripts. In contrast, no changes in plasma glucose, circulating insulin, pancreatic insulin content, or insulin messenger RNA were observed in mice, 18 h after administration of GIPR Ab. These findings demonstrate that GLP-1, but not GIP, plays an essential role in regulating glycemia, independent of enteral nutrient ingestion in mice in vivo. PMID- 11014226 TI - Glucose-dependent insulinotropic polypeptide confers early phase insulin release to oral glucose in rats: demonstration by a receptor antagonist. AB - A novel GIP receptor antagonist was developed to evaluate the acute role of glucose-dependent insulinotropic polypeptide (GIP) in the insulin response to oral glucose in rats. Antisera to an extracellular epitope of the GIP receptor (GIPR) detected immunoreactive GIPR on rat pancreatic beta-cells. Purified GIPR antibody (GIPR Ab) specifically displaced GIP binding to the receptor and blocked GIP-mediated increases in intracellular cAMP. When delivered to rats by ip injection, GIPR Ab had a half-life of approximately 4 days. Treatment with GIPR Ab (1 microg/g BW) blocked the potentiation of glucose-stimulated insulin secretion by GIP (60 pmol) but not glucagon-like peptide-1 (GLP-1, 60 pmol) in anesthetized rats. The insulin response to oral glucose was delayed in conscious unrestrained rats that were pretreated with GIPR Ab. Plasma insulin levels were approximately 35% lower at 10 min in GIPR Ab treated animals compared with controls. As a result, the glucose excursion was greater in the GIPR Ab treated group. Fasting plasma glucose levels were not altered by GIPR Ab. We conclude that release of GIP following oral glucose may act as an anticipatory signal to pancreatic beta-cells to promote rapid release of insulin for glucose disposal. PMID- 11014227 TI - Juxtamembrane regions in the third intracellular loop of the thyrotropin releasing hormone receptor type 1 are important for coupling to Gq. AB - Juxtamembrane residues in the putative third intracellular (I3) loops of a number of G protein-coupled receptors (GPCRs) have been shown to be important for coupling to G proteins. According to standard hydropathy analysis, the I3 loop of the mouse TRH receptor type 1 (mTRH-R1) is composed of 51 amino acids from position-213 to position-263. We constructed deletion and site-specific I3 loop TRH-R mutants and studied their binding and TRH-stimulated signaling activities. As expected, the effects of these mutations on TRH binding were small (less than 5-fold decreases in affinity). No effect on TRH-stimulated signaling activity was found in a mutant receptor in which the I3 loop was shortened to 16 amino acids by deleting residues from Asp-226 to Ser-260. In contrast, mutants with deletions from Asp-222 to Ser-260 or from Asp-226 to Gln-263 exhibited reduced TRH stimulated signaling. In the region near transmembrane helix 6, single site specific substitution of either Arg-261 or Lys-262 by neutral glutamine had little effect on signaling, but mutant TRH-Rs that were substituted by glutamine at both basic residues exhibited reduced TRH-stimulated activity. The reduced signaling activity of this doubly substituted mutant was reversed by over expressing the a subunit of Gq. These data demonstrate that the juxtamembrane regions in the TRH-R I3 loop are important for coupling to Gq. PMID- 11014228 TI - Pituitary adenylate cyclase-activating polypeptide precursor is processed solely by prohormone convertase 4 in the gonads. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is abundant not only in the brain, but also in the testis. Immunohistochemical studies have shown that PACAP-LI in rat testis is expressed stage specifically in spermatids. This suggests that testicular PACAP participates in the regulatory mechanism of spermatogenesis. Additionally, the ovary contains a relatively small amount of PACAP, conceivably involved in the regulation of folliculogenesis. PACAP is synthesized as a preprohormone and is processed by prohormone convertases, such as PC1, PC2, and PC4. PC4 is expressed only in the testis and ovary, where neither PC1 nor PC2 is expressed. However, whether PC4 is the sole endoprotease for the PACAP precursor in the gonads remains unknown. Recent studies using PC4 transgenic mice revealed that male PC4-null mice exhibited severely impaired fertility, although spermatogenesis appeared to be normal. The female PC4-null mice exhibited delayed folliculogenesis in the ovaries. To examine whether PC4 is the sole processing enzyme for the PACAP precursor in the gonads, we analyzed testicular and ovarian extracts from the PC4-null and wild-type mice for PACAP (PACAP38 and PACAP27) and its messenger RNA using reverse phase HPLC combined with specific RIAs and ribonuclease protection assay, respectively. For RIAs, three different polyclonal antisera with different recognition sites were used to identify PACAP38, PACAP27, and its precursor. Neither the testis nor the ovary from the PC4-null mice expressed PACAP38 or PACAP27, but the levels of PACAP transcripts in the testis and ovary of homozygous PC4-deficient mice were considerably elevated compared with those of the wild-type and heterozygous animals. The findings indicate that PC4 is the sole processing enzyme for the precursor of PACAP in the testis and ovary of mice. The possibility that the absence of bioactive PACAP in the testis and ovary of PC4-null mice caused severely impaired fertility in the males and delayed folliculogenesis in females warrants investigation. PMID- 11014229 TI - Whole-cell recordings from preoptic/hypothalamic slices reveal burst firing in gonadotropin-releasing hormone neurons identified with green fluorescent protein in transgenic mice. AB - Central control of reproduction is governed by a neuronal pulse generator that underlies the activity of hypothalamic neuroendocrine cells that secrete GnRH. Bursts and prolonged episodes of repetitive action potentials have been associated with hormone secretion in this and other neuroendocrine systems. To begin to investigate the cellular mechanisms responsible for the GnRH pulse generator, we used transgenic mice in which green fluorescent protein was genetically targeted to GnRH neurons. Whole-cell recordings were obtained from 21 GnRH neurons, visually identified in 200-microm preoptic/hypothalamic slices, to determine whether they exhibit high frequency bursts of action potentials and are electrically coupled at or near the somata. All GnRH neurons fired spontaneous action potentials, and in 15 of 21 GnRH neurons, the action potentials occurred in single bursts or episodes of repetitive bursts of high frequency spikes (9.77 +/- 0.87 Hz) lasting 3-120 sec. Extended periods of quiescence of up to 30 min preceded and followed these periods of repetitive firing. Examination of 92 GnRH neurons (including 32 neurons that were located near another green fluorescent protein-positive neuron) revealed evidence for coupling in only 1 pair of GnRH neurons. The evidence for minimal coupling between these neuroendocrine cells suggests that direct soma to soma transfer of information, through either cytoplasmic bridges or gap junctions, has a minor role in synchronization of GnRH neurons. The pattern of electrical activity observed in single GnRH neurons within slices is temporally consistent with observations of GnRH release and multiple unit electrophysiological correlates of LH release. Episodes of burst firing of individual GnRH neurons may represent a component of the GnRH pulse generator. PMID- 11014231 TI - Selective involvement of interleukin-6 in the transcriptional activation of the suppressor of cytokine signaling-3 in the brain during systemic immune challenges. AB - Cytokine-inducible proteins named as suppressors of cytokine signaling (SOCS) are rapidly induced by interleukin-6 (IL-6) and other members sharing the gp130 receptor subunit after activation of the Janus kinases (JAK) and the signal transducers and activators of transcription (STAT). These inhibitory proteins generally prevent tyrosine phosphorylation of IL-6 receptor signaling subunit gp130, specific JAK and STAT or in acting at steps distal to JAK activation. Expression of these inhibitory proteins is therefore a useful tool to investigate the signaling events occurring in the brain during immunogenic stimuli that involve cytokines of the IL-6 family. This study investigated the effect of ip lipopolysaccharide (LPS) administration on the expression of one key member of the SOCS family, SOCS-3, in both rats and mice. In rats, the endotoxin caused a profound transcriptional activation of the inhibitory factor in the circumventricular organs subfornical organ, organum vasculosum of the lamina terminalis, arcuate nucleus/median eminence, area postrema, choroid plexus, leptomeninges, ependymal lining cells, and along the endothelium of the brain blood vessels. The hybridization signal for SOCS-3 messenger RNA was low at 1 h, but robust at 3 and 6 h and declined to return to basal levels 12 h after the single ip LPS injection. The pattern of SOCS-3 expression was similar in the brain of wild-type mice, although induction of the inhibitory factor was no longer observed in the ependymal lining cells of the cerebral ventricles and the blood microvessels of IL-6-deficient animals at all the times evaluated, i.e. from 1-8 h post-LPS injection. The endothelium of the brain capillaries also exhibited up-regulation of both IL-6 receptor and gp130 subunits during systemic inflammation, which allowed SOCS-3 expression in response to circulating IL-6. The present data indicate that the JAK/STAT transduction pathways that lead to SOCS-3 transcription are activated within cells accessible from the blood circulation, but not within deep parenchymal elements of the brain during endotoxemia. Induction of SOCS-3 followed the cascade of events that take place during the acute phase response and the contribution of IL-6 in activating the inhibitory factor is site specific and not generalized throughout the central nervous system. PMID- 11014230 TI - Promoter elements and transcription factors involved in differentiation-dependent human chorionic gonadotrophin-alpha messenger ribonucleic acid expression of term villous trophoblasts. AB - Differentiation of primary villous cytotrophoblasts into syncytia is associated with increasing production of alpha and beta human CG subunits, which is predominantly governed at the level of messenger RNA expression. Here, we present a detailed study on the mechanisms involved in the differentiation-dependent regulation of the trophoblast-specific CGalpha gene promoter. Site-directed mutations in each of the five DNA-elements of the composite enhancer were performed to investigate the contribution of the individual regulatory sequences to the overall transcriptional activity of the promoter at two different stages of trophoblast in vitro differentiation. We show that deletion of one cyclic AMP response element (CRE) did not affect CGalpha promoter activity in cytotrophoblasts; however, it reduced transcription by 33% in differentiating cultures. Removal of both CREs almost abolished transcription at early and later stages of in vitro differentiation. Upon mutation the enhancer elements alphaACT, JRE, and CCAAT significantly decreased luciferase reporter transcription; however their contribution to the total promoter activity did not change during in vitro differentiation. Contrary to that, mutated TSE diminished promoter activity by 19% during 12 and 48 h of cultivation but reduced luciferase expression by 78% between 48 and 84 h of differentiation. In electrophoretic mobility shift assay, the TSE interacted with activating protein (AP)-2alpha in both primary trophoblasts and choriocarcinoma cells. While CRE-interacting proteins were detectable 12 h after isolation, the TSE-binding complex did not appear before 36 h of in vitro differentiation. During syncytium formation increasing protein expression of activating transcription factor (ATF)-1, cAMP response element binding protein (CREB)-1, and AP-2alpha was observed on Western blots. Moreover, phosphorylated CREB-1 and ATF-1 accumulated between 24 and 78 h of trophoblast cultivation. By fluorescence immunohistochemistry, we show that CREB-1 was predominantly expressed in syncytiotrophoblasts, whereas ATF-1 and AP-2alpha localized to the syncytium and some cytototrophoblasts as well as to stromal and endothelial cells of the placental villus. Phosphorylated CREB-1/ATF-1 and the coactivator protein CBP were primarily detected in syncytial nuclei, suggesting the presence of functional, cAMP-dependent transcriptional complexes in the differentiated tissue. In agreement to the in vivo situation, phosphorylated CREB 1/ATF-1 were observed in nuclei of the differentiated trophoblast cultures. The activity of the CGalpha promoter as well as CREB-1/ATF-1 phosphorylation increased upon elevation of cAMP levels and overexpression of the catalytic subunit of protein kinase A. Additionally, we demonstrate that overproduction of the enzyme enhanced protein expression and binding of AP-2alpha to the TSE. We conclude that differentiation-dependent transcription of the CGalpha gene in villous trophoblasts is mainly governed by increasing expression of AP-2alpha and PKA-dependent phosphorylation of CREB-1 and ATF-1. PMID- 11014232 TI - Functional significance of MMP-9 in tumor necrosis factor-induced proliferation and branching morphogenesis of mammary epithelial cells. AB - Tissue remodeling is a key process involved in normal mammary gland development, with matrix metalloproteinases (MMPs) playing an important role in this process. Our laboratory has demonstrated that tumor necrosis factor (TNF) stimulates branching morphogenesis of mammary epithelial cells (MEC) within a reconstituted basement membrane. Studies were therefore undertaken to determine whether MMPs might mediate the effects of TNF. Using a primary culture model in which rat MEC grow three-dimensionally within a reconstituted basement membrane, we found that TNF stimulated secretion of MMP-9 but not MMP-2. To determine whether MMP-9 was involved in TNF-induced proliferation and branching morphogenesis, we used a peptide containing the prodomain sequence of MMPs and two MMP inhibitors. Both the prodomain peptide (5 x 10(-4)-10(-3) M), as well as BB-94 (10(-8)-10(-5) M) and CGS 27023A (10(-6)-10(-5) M), inhibited TNF-induced proliferation and branching morphogenesis in a concentration-dependent manner. Finally, to verify the specific requirement for MMP-9, we demonstrated that an MMP-9 neutralizing antibody blocked TNF-induced proliferation and branching morphogenesis. Together, these data suggest that TNF-regulated MMP-9 may play a role in the controlled invasion of the fad pad that occurs during normal mammary gland development and that misregulation of MMP-9 may contribute to the invasiveness of breast cancer. PMID- 11014233 TI - Calcitonin receptor regulation and responsiveness to calcitonin in human osteoclast-like cells prepared in vitro using receptor activator of nuclear factor-kappaB ligand and macrophage colony-stimulating factor. AB - Using mouse osteoclast-like cells (OCs), we have shown that short exposure to calcitonin (CT) resulted in prolonged reduction of CT binding by inhibiting de novo CT receptor (CTR) synthesis. Additionally, CT-treated OCs demonstrated resistance to CT rechallenge on the inhibitory effect of CT in osteoclastic bone resorption. There is, however, scant information on CT effects on human osteoclasts. In this study, we examined the features of CTR down-regulation and its recovery after short exposure to CT of human OCs. OCs were prepared by treatment of peripheral blood mononuclear cells in vitro with osteoclast differentiation factor and macrophage colony-stimulating factor. Treatment of OCs with salmon CT (sCT) and human CT (hCT) resulted in a dose-dependent reduction in [125I]sCT binding capacity. Continued receptor occupancy by ligand was excluded by using a glycine-acid washing procedure. Treatment with sCT reduced CTR messenger RNA expression, suggesting that CTR down-regulation is, at least partly, attributable to an inhibition of de novo CTR synthesis. To investigate the intracellular signal transduction pathways that mediate these effects, we examined the effects of activation of the protein kinase (PK)A, PKC, and Ca2+ calmodulin-dependent kinases. Treatment with PKC activators mimicked CT, whereas neither activation of PKA nor elevation of intracellular Ca2+ did so. We further investigated the intracellular signaling pathways responsible for the inhibitory effects of CT on bone resorption, which showed that treatment with PKC activators reproduced the effects of CT. These data suggest that the PKC pathway plays an important role in homologous CTR down-regulation, as well as inhibition of bone resorbing activity by CT, in human OCs. Short exposure of OCs to CT (10(-9) M, 1 h) reduced [125I]sCT-specific binding for a prolonged period, as we have shown previously in mouse OCs. The reduced specific binding, CTR messenger RNA levels, and CT-sensitive adenylate cyclase responsiveness returned to the control levels by 96 h after removal of CT. These results strongly support the notion that escape from CT inhibition of osteoclastic bone resorption in humans is attributable to the development of resistance by OCs to CT. This study also showed that even short exposure to CT induced prolonged desensitization to CT rechallenge, although the OCs eventually regained responsiveness to sCT rechallenge. PMID- 11014234 TI - Prostaglandin production at the onset of ovine parturition is regulated by both estrogen-independent and estrogen-dependent pathways. AB - A current hypothesis of ovine parturition proposes that fetal adrenal cortisol induces placental E2 production, which, in turn, triggers intrauterine PG production. However, recent evidence suggests that cortisol may directly increase PG production in trophoblast-derived tissues. To separate cortisol-dependent and estrogen-dependent PG production in sheep intrauterine tissues, we infused singleton, chronically catheterized fetuses beginning on day 125 of gestation (term, 147-150 days) with 1) cortisol (1.35 mg/h; n = 5); 2) cortisol and 4 hydroxyandrostendione, a P450aromatase inhibitor (4-OHA: 1.44 mg/h; n = 5); 3) saline (n = 5); or 4) saline and 4-OHA (n = 5). Fetal and maternal arterial blood samples were collected at 12-h intervals starting 24 h before infusion and continuing during treatment for 80 h or until active labor. Uterine contractility was measured by electromyogram recording of myometrial activity. Plasma E2, progesterone (P4), PGE2, and 13,14-dihydro- 15-keto-PGF2alpha were quantified by RIA. PGHS-II messenger RNA (mRNA) and protein expression were determined by in situ hybridization and Western blot analysis, respectively. Data were analyzed by ANOVA (P < or = 0.05). Labor-type uterine contractions were present after 68 h of cortisol infusion and had increased significantly by 80 h. Labor-type uterine contractions were induced after 68 h of cortisol plus 4-OHA infusion, but the contraction frequency remained less than that in the cortisol-treated animals. Fetal cortisol infusion increased fetal and maternal plasma E2 concentrations and decreased the maternal plasma P4 concentration significantly; concurrent 4-OHA infusion attenuated the increase in fetal and maternal plasma E2, but not the decrease in maternal plasma P4. The fetal plasma PGE2 concentration increased after both cortisol and cortisol plus 4-OHA infusion. The maternal plasma 13,14 dihydro-15-keto-PGF2alpha concentration rose after fetal cortisol infusion, but not after cortisol plus 4-OHA infusion. Placental trophoblast PGHS-II mRNA and protein expression were increased significantly after both cortisol and cortisol plus 4-OHA infusion. Endometrial PGHS-II mRNA and protein expression increased after cortisol infusion, but not after cortisol plus 4-OHA infusion. Plasma steroid and PG concentrations, uterine activity pattern, and intrauterine PGHS-II expression were not altered in either control group. We conclude that these data suggest distinct pathways of intrauterine PG synthesis: a cortisol-dependent/E2 independent mechanism within trophoblast tissue leading to elevations in fetal plasma PGE2, and an E2-dependent mechanism within maternal endometrium that leads to increased maternal plasma PGF2alpha and appears necessary for uterine activity and parturition. PMID- 11014235 TI - The differential fate of mesonephric tubular-derived efferent ductules in estrogen receptor-alpha knockout versus wild-type female mice. AB - We investigated mesonephric tubular-derived efferent ductules in female wild-type (WT) and estrogen receptor-alpha knockout (ERalphaKO) mice from late fetal to adult life. On gestational day 17, efferent ductules in both fetal WT and ERalphaKO females were well developed and morphologically similar, although one third the size of the male counterpart. Unexpectedly, efferent ductules with a ciliated epithelium were still present on postnatal day 10 in WT and ERalphaKO females. By day 23, however, marked phenotypic differences occurred in efferent ductules of WT and ERbetaKO vs. ERalphaKO female mice. In the latter, efferent ductules became hypertrophied and dilated, whereas only small tubules remained in WT and ERbetaKO adult mice. The serum testosterone concentrations were similar in 21- to 25-day-old ERalphaKO, heterozygous, and WT female mice, suggesting that increased testosterone was not inducing enlargement of efferent ductules in ERalphaKO females. In conclusion, remnants of efferent ductules persisted in normal adult female mice, although these structures were greatly reduced in size compared with efferent ductules in ERalphaKO female mice. The underlying mechanism inducing hypertrophy and dilation of efferent ductules in ERalphaKO females is not clear, but secretory and/or reabsorptive function of female efferent ductules may involve ERalpha. PMID- 11014236 TI - Circadian and glucocorticoid regulation of Rev-erbalpha expression in liver. AB - Rev-erbalpha [NR1D1], a member of the nuclear receptor superfamily, is an orphan receptor that constitutively represses gene transcription. Rev-erbalpha has been shown to play a role in myocyte differentiation and to be induced during adipogenesis. Furthermore, Rev-erbalpha is a regulator of lipoprotein metabolism. It was recently shown that Rev-erbalpha messenger RNA (mRNA) levels oscillate diurnally in rat liver. Here, we report that the circadian rhythm of Rev-erbalpha in liver is maintained in primary cultures of rat hepatocytes. Because glucocorticoids have been shown to regulate other transcription factors with circadian expression, it was furthermore examined whether hepatic Rev-erbalpha expression is also regulated by glucocorticoids. Treatment of rats with dexamethasone resulted in a decrease of Rev-erbalpha mRNA levels by 70% after 6 h. Furthermore, dexamethasone decreased Rev-erbalpha expression in rat primary hepatocytes in a dose-dependent fashion. This effect was mediated by the glucocorticoid receptor because simultaneous addition of the glucocorticoid antagonist RU486 prevented the decrease in Rev-erbalpha mRNA levels by dexamethasone. Protein synthesis inhibition with cycloheximide markedly induced Rev-erbalpha mRNA levels; however, this induction was reduced by dexamethasone supplementation in both rat and human primary hepatocytes. Treatment with actinomycin D blocked the repression of Rev-erbalpha expression by dexamethasone in rat hepatocytes, suggesting that glucocorticoids regulate Rev-erbalpha expression at the transcriptional level. Transient transfection experiments further indicated that Rev-erbalpha promoter activity is repressed by dexamethasone in the presence of cotransfected glucocorticoid receptor. Taken together, these data demonstrate that Rev-erbalpha expression is under the control of both the circadian clock and glucocorticoids in the liver. PMID- 11014237 TI - Genetic models reveal that brain natriuretic peptide can signal through different tissue-specific receptor-mediated pathways. AB - Brain natriuretic peptide (BNP), a hormone produced primarily by the cardiac ventricle, is thought to be involved in a variety of homeostatic processes through its cognate receptor, guanylyl cyclase A (GC-A). We previously created transgenic mice overexpressing BNP under the control of the liver-specific human serum amyloid P component promoter (BNP-transgenic mice) and demonstrated that they exhibit reduced blood pressure and cardiac weight accompanied by an elevation of plasma cGMP concentrations and marked skeletal overgrowth through the activation of endochondral ossification. To address whether BNP exerts its biological effects solely through GC-A, we produced BNP-transgenic mice lacking GC-A (BNP-Tg/GC-A-/- mice) and examined their cardiovascular and skeletal phenotypes. The GC-A-/- mice are hypertensive with cardiac hypertrophyrelative to wild-type littermates, which is not alleviated by overexpression of BNP in BNP Tg/GC-A-/- mice. The BNP-Tg/GC-A-/- mice, however, continue to exhibit marked longitudinal growth of vertebrae and long bones comparably to BNP-Tg mice. This study provides genetic evidence that BNP reduces blood pressure and cardiac weight through GC-A, whereas it dramatically alters endochondral ossification in the absence of this receptor. Therefore, the BNP-Tg/GC-A-/- mice provide the first experimental model demonstrating that this natriuretic peptide can signal in a tissue-specific manner through a receptor other than GC-A. PMID- 11014238 TI - In vivo treatment with GDF-9 stimulates primordial and primary follicle progression and theca cell marker CYP17 in ovaries of immature rats. AB - Growth differentiation factor (GDF)-9 is a cystine knot-containing hormone of the transforming growth factor-beta superfamily produced by the oocyte. In GDF-9 null mice, follicle development is arrested at the primary stage and GDF-9 treatment in vitro enhances preantral follicle growth. Immature female rats were treated with recombinant GDF-9 for 7 or 10 days. At 10 days, treatment with GDF-9 augmented ovarian weights, concomitant with an increase in the number of primary and small preantral follicles by 30 and 60%, respectively. Furthermore, the number of primordial follicles was decreased by 29%, but the number of large preantral follicles was not affected. In contrast, treatment with FSH increased the number of small and large preantral follicles by 36 and 177% but did not influence the number of primary and primordial follicles. Immunoblot analysis showed an increase of CYP17, a theca cell marker, in the ovarian homogenate after treatment with GDF-9 but not FSH. The present results indicate that in vivo treatment with GDF-9 enhances the progression of primordial and primary follicles into small preantral follicles. Thus, GDF-9 treatment could provide an alternative approach to stimulate early follicle development in addition to the widely used FSH that acts mainly on the development of more advanced follicles. PMID- 11014239 TI - A novel messenger ribonucleic acid homologous to human MAGE-D is strongly expressed in rat Sertoli cells and weakly in Leydig cells and is regulated by follitropin, lutropin, and prolactin. AB - We have cloned a novel complementary DNA whose expression was decreased in rat Sertoli cell cultures after treatment with FSH. This complementary DNA encodes a protein of 570 amino acids and shares 92% homology with the human MAGE-D protein. In contrast to other MAGE genes (A, B, or C), we have shown that MAGE-D expression was ubiquitous in healthy rat tissues. In the seminiferous tubules, the MAGE-D was expressed in Sertoli cells but not in germ cells as demonstrated by RT-PCR and in situ hybridization, whereas for the other MAGE genes, expression has been shown to be restricted to germ cells. Interestingly, MAGE-D was also detected for the first time in the female gonad by Northern blotting. In MLTC-1 cells (mouse Leydig tumor cell line-1), LH and PRL stimulated MAGE-D expression. Using hypophysectomized rats, it was confirmed that FSH decreased MAGE-D expression, whereas LH and PRL increased MAGE-D messenger RNA level in the whole testis most probably through a direct action on Leydig cells. As MAGE-D is present in both the seminiferous compartment and interstitium and hormonally regulated in each, it is possible that it has specific functions in each compartment during the development and the maintenance of the testis. PMID- 11014240 TI - Functional connections between the suprachiasmatic nucleus and the thyroid gland as revealed by lesioning and viral tracing techniques in the rat. AB - Frequent blood sampling via intraatrial cannula revealed daily rhythms of TSH and thyroid hormones in both male and female Wistar rats. Thermic ablation of the biological clock, i.e. the suprachiasmatic nucleus (SCN), eliminated the diurnal peak in circulating TSH and thyroid hormones. In addition, SCN lesions produced a clear decrease of 24-h mean T4 concentrations. A more pronounced effect of SCN lesions on thyroid hormones, as opposed to TSH, suggested routes of SCN control additional to the neuroendocrine hypothalamopituitary-thyroid axis. Retrograde, transneuronal virus tracing was used to identify the type and localization of neurons in the central nervous system that control the autonomic innervation of the thyroid gland. In the spinal cord and brain stem, both the sympathetic and parasympathetic motorneurons were labeled. By varying the postinoculation survival time, it was possible to follow the viral infection as it proceeded. Subsequently, the pseudorabies virus (PRV) infected neurons in several brain stem cell groups, the paraventricular nucleus of the hypothalamus (PVN) and the central nucleus of the amygdala (second order labeling). Among PRV-infected neurons in the PVN were TRH-containing cells. Third order neurons were found in several hypothalamic cell groups, among which was the SCN. Therefore, we propose that the SCN has a dual control mechanism for thyroid activity by affecting neuroendocrine control of TSH release on the one hand and the autonomic input to the thyroid gland on the other. PMID- 11014241 TI - Estrogen receptors alpha and beta in rat decidua cells: cell-specific expression and differential regulation by steroid hormones and prolactin. AB - Estradiol is known to play an important role in the growth and differentiation of rat uterine stromal cells into decidual cells. In particular, this hormone with progesterone is necessary for blastocyst implantation and subsequent decidualization in the rat. Although binding experiments have demonstrated the presence of estrogen-binding sites, no evidence exists as to whether the rat decidua expresses both isoforms of the estrogen receptor (ER), alpha and beta. In this investigation, we analyzed the expression of decidual ERalpha and ERbeta, studied their regulation by PRL and steroid hormones and examined the ability of decidual ERp to transduce the estradiol signal to the progesterone receptor. Immunocytochemistry, RT-PCR, and Northern blot analysis showed that both ER species are coexpressed in the decidua during pseudopregnancy. Interestingly, these genes were preferentially found in a cell population localized in the antimesometrial site of the uterus where blastocyst implantation takes place. Using decidual cells in primary culture obtained from pseudopregnant rats and a decidua-derived cell line (GG-AD), we show a differential regulation of ERalpha and ERbeta by PRL and ovarian steroid hormones. Whereas PRL, estradiol, and progesterone all increased ERbeta messenger RNA (mRNA) expression in a dose dependent manner, only PRL up-regulated the mRNA levels of ERa. Estradiol had no effect on ERalpha expression, whereas progesterone markedly decreased its mRNA levels. Interestingly, progesterone, which up-regulates the ability of PRL to signal to a PRL-regulated gene in mammary-gland derived cells, prevented PRL stimulation of decidual ERalpha and had no synergistic effect on ERbeta expression. The use of GG-AD cells, which express only ERbeta, allowed us to demonstrate that this receptor subtype is functional and transduces estradiol signal to the progesterone receptor. In summary, the results of this investigation have revealed that ERbeta is expressed in addition to ERalpha in the rat decidua, and that the expression of both ERs are cell specific and differentially regulated by PRL and steroids. One salient finding of this investigation is that progesterone down-regulates ERalpha, but concomitantly increases the expression of a functional ERbeta that mediates estradiol up regulation of the decidual progesterone receptor. PMID- 11014242 TI - In situ estrogen synthesized by aromatase P450 in uterine leiomyoma cells promotes cell growth probably via an autocrine/intracrine mechanism. AB - In the present study we characterized in detail the expression of aromatase P450 in leiomyomas to determine the role of in situ estrogen in the growth advantage of leiomyomas. The levels of aromatase P450 transcripts were determined by quantitative RT-PCR to be significantly higher in leiomyomas than in corresponding myometrium. The overexpression of aromatase P450 in leiomyomas was also confirmed by Western blot analysis. The estimated size of immunoreactive aromatase was 58 kDa, similar to that in placenta. To identify a cell type that express aromatase P450 in leiomyomas, histological specimens were stained for aromatase P450 using a polyclonal antibody. Strong immunoreactivity was detected in the cytoplasm of leiomyoma cells, whereas surrounding normal myometrium displayed weak or negative staining. Smooth muscle-like cells in culture obtained from leiomyomas, positive for actin D fiber, possessed immunoreactive granules of aromatase in the cytoplasm. Conversion of androgen to estrogen was effectively stimulated by phorbol myristate acetate and dexamethasone plus interleukin-1beta and was completely abolished by selective inhibitors of aromatase P450 (fadrozole and TZA-2209), but not by inhibitors of 5alpha-reductase (finasteride and flutamide). The apparent Km of androstenedione was 3 nM in the presence of dexamethasone and interleukin-1beta, corresponding to the plasma concentration of androstenedione in women of reproductive age. To determine whether endogenous aromatase P450 plays a role in the growth promotion of leiomyoma cells, we evaluated the cell growth of smooth muscle-like cells treated with various concentrations of estrogen and androgen using a WST-1 assay. Treatment with testosterone (10(-8) and 10(-7) M) and androstenedione (10(-8) and 10(-7) M) stimulated the growth of smooth muscle-like cells obtained from leiomyomas to the same extent as estradiol (10(-10)-10(-7) M), whereas dihydrotestosterone (10(-11) 10(-8) M) did not. The stimulatory effect of testosterone on cell growth was again abolished by cotreatment with fadrozole. The level of estradiol in the medium of testosterone (10(-8) M)-treated smooth muscle-like cells was 10(-11) M, which was 1 order lower than the minimum concentration of estradiol necessary to promote cell growth (10(-10) M). This indicates that estradiol synthesized in leiomyomas promotes their growth via an autocrine/intracrine mechanism. We conclude that myometrial cells of leiomyomas overexpress aromatase P450 and are able to synthesize sufficient estrogen to accelerate their own cell growth. Overexpression of aromatase P450 may play a role in the growth advantage of leiomyoma tissue over surrounding myometrium via an autocrine/intracrine mechanism. PMID- 11014243 TI - The role of D-aspartic acid and N-methyl-D-aspartic acid in the regulation of prolactin release. AB - In this study, using an enzymatic HPLC method in combination with D-aspartate oxidase, we show that N-methyl-D-aspartate (NMDA) is present at nanomolar levels in rat nervous system and endocrine glands as a natural compound, and it is biosynthesized in vivo and in vitro. D-aspartate (D-Asp) is its natural precursor and also occurs as an endogenous compound. Among the endocrine glands, the highest quantities of D-Asp (78 +/- 12 nmol/g) and NMDA (8.4 +/- 1.2 nmol/g) occur in the adenohypophysis, whereas the hypothalamus represents the area of the nervous system where these amino acids are most abundant (55 +/- 9 and 5.6 +/- 1.1 nmol/g for D-Asp and NMDA, respectively). When D-Asp is administered to rats by ip injection, there is a significant uptake of D-Asp into the adenohypophysis and a significant increase in the concentration of NMDA in the adenohypophysis, hypothalamus and hippocampus, suggesting that D-Asp is an endogenous precursor for NMDA biosynthesis. Experiments conducted on tissue homogenates confirm that D Asp is the precursor of the NMDA and that the enzyme catalyzing this reaction is a methyltransferase. S-adenosyl-L-methionine (SAM) is the methyl group donor. In vivo experiments consisting of ip injections of sodium D-aspartate show that this amino acid induced a significant serum PRL elevation and this effect is dose and time dependent. In vitro experiments conducted on isolated adenohypophysis or adenohypophysis coincubated with the hypothalamus, showed that the release of PRL is caused by a direct action of D-Asp on the pituitary gland and also mediated by the indirect action of NMDA on the hypothalamus. Then, the latter induces the release of a putative factor that in turn stimulates the adenohypophysis reinforcing the PRL release. In conclusion, our data suggest that D-Asp and NMDA are present endogenously in the rat and are involved in the modulation of PRL release. PMID- 11014244 TI - Aldosterone: a mediator of myocardial necrosis and renal arteriopathy. AB - To determine the role of aldosterone in mediating cardiovascular damage, we performed ablation/replacement experiments with aldosterone in a rat model of cardiac injury. Administration of angiotensin II and Nomega-nitro-L-arginine methyl ester (L-NAME; nitric oxide synthesis inhibitor) to male rats drinking 1% saline caused hypertension, severe biventricular myocardial necrosis, proteinuria, and fibrinoid necrosis of renal and cardiac vessels. Removal of aldosterone by adrenalectomy or through administration of the selective aldosterone antagonist eplerenone markedly reduced the cardiac and renal damage without significantly altering blood pressure. Aldosterone infusion in adrenalectomized, glucocorticoid-replaced L-NAME/angiotensin II-treated animals restored damage. Thus, we identified aldosterone as a critical mediator of L NAME/angiotensin II induced vascular damage through mechanisms apparently independent of its effects on systolic blood pressure. PMID- 11014245 TI - Growth hormone increases connexin-43 expression in the cerebral cortex and hypothalamus. AB - Several studies indicate that systemic GH influences various brain functions. Connexin-43 forms gap junctions that mediate intercellular communication and establish the astroglial syncytium. We investigated the effects of peripheral administration of bovine GH (bGH) and recombinant human insulin-like growth factor I (rhIGF-I) on the expression of connexin-43 in the rat brain. Hypophysectomized female Sprague Dawley rats were substituted with cortisol (400 microg/kg x day) and L-T4 (10 microg/kg x day) and treated with either bGH (1 mg/kg x day) or rhIGF-I (0.85 mg/kg x day) for 19 days. The abundance of connexin 43 messenger RNA (mRNA) and protein in the brainstem, cerebral cortex, hippocampus, and hypothalamus was quantified by means of ribonuclease protection assays and Western blots. Treatment with bGH increased the amounts of connexin-43 mRNA and protein in the cerebral cortex and hypothalamus. No changes were found in the brainstem or hippocampus. Infusion of rhIGF-I did not affect connexin-43 mRNA or protein levels in any of the brain regions studied. These results show that administration of bGH increases the abundance of cx43 in specific brain regions, suggesting that GH may influence gap junction formation and thereby intercellular communication in the brain. PMID- 11014246 TI - Thyroid hormone acts directly on growth plate chondrocytes to promote hypertrophic differentiation and inhibit clonal expansion and cell proliferation. AB - T3 is an important regulator of endochondral bone formation in epiphyseal growth plates. Growth arrest in juvenile hypothyroidism results from disorganization of growth plate chondrocytes and their failure to undergo hypertrophic differentiation, but it is unclear how T3 acts directly on chondrocytes or whether its actions involve other pathways. To address this issue, we investigated whether thyroid hormone receptors (TR) were localized to discrete regions of the unfused epiphysis by immunohistochemistry performed in tibial growth plates from 21-day-old rats and examined the effects of T3 on growth plate chondrocytes in agarose suspension cultures in vitro. TRalpha1, -alpha2, and beta1 were expressed in reserve and proliferating zone chondrocytes, but not in hypertrophic cells, suggesting that progenitor cells and immature chondrocytes are the major T3 target cells in the growth plate. Chondrocytes in suspension culture expressed TRalpha1, -alpha2, and -beta1 messenger RNAs and matured by an ordered process of clonal expansion, colony formation, and terminal hypertrophic differentiation. Clonal expansion and proliferation of chondrocytes were inhibited by T3, which also induced alkaline phosphatase activity, expression of collagen X messenger RNA, and secretion of an alcian blue-positive matrix as early as 7 days after hormone stimulation. Thus, T3 inhibited chondrocyte clonal expansion and cell proliferation while simultaneously promoting hypertrophic chondrocyte differentiation. These data indicate that thyroid hormones concurrently and reciprocally regulate chondrocyte cell growth and differentiation in the endochondral growth plate. PMID- 11014247 TI - Comparative effects of neonatal exposure of male rats to potent and weak (environmental) estrogens on spermatogenesis at puberty and the relationship to adult testis size and fertility: evidence for stimulatory effects of low estrogen levels. AB - This study investigated whether neonatal exposure of male rats to estrogenic compounds altered pubertal spermatogenesis (days 18 and 25) and whether the changes observed resulted in long-term changes in testis size, mating, or fertility (days 90-100). Rats were treated neonatally with a range of doses (0.01 10 microg) of diethylstilbestrol (DES; administered on alternate days from days 2 12), a high dose of octylphenol (OP; 2 mg administered daily from days 2-12) or bisphenol A (Bis-A; 0.5 mg administered daily from days 2-12), or vehicle, while maintained on a standard soy-containing diet. The effect on the same parameters of rearing control animals on a soy-free diet was also assessed as was the effect of administering such animals genistein (4 mg/kg/day daily from days 2-18). Testis weight, seminiferous tubule lumen formation, the germ cell apoptotic index (apoptotic/viable germ cell nuclear volume), and spermatocyte nuclear volume per unit Sertoli cell nuclear volume were used to characterize pubertal spermatogenesis. Compared with (soy-fed) controls, DES administration caused dose dependent retardation of pubertal spermatogenesis on day 18, as evidenced by decreases in testis weight, lumen formation, and spermatocyte nuclear volume per unit Sertoli cell and elevation of the germ cell apoptotic index. However, the two lowest doses of DES (0.1 and 0.01 microg) significantly increased spermatocyte nuclear volume per unit Sertoli cell. Similarly, treatment with either OP or Bis-A significantly advanced this and some of the other aspects of pubertal spermatogenesis. Maintenance of control animals on a soy-free diet also significantly advanced lumen formation and spermatocyte nuclear volume per unit Sertoli cell compared with controls fed a soy-containing diet. Administration of genistein reversed the stimulatory effects of a soy-free diet and significantly retarded most measures of pubertal spermatogenesis. In general, plasma FSH levels in the treatment groups changed in parallel to the spermatogenic changes (reduced when pubertal spermatogenesis retarded, increased when pubertal spermatogenesis advanced). By day 25, although the changes in FSH levels largely persisted, all of the stimulatory effects on spermatogenesis seen on day 18 in the various treatment groups were no longer evident. In adulthood, testis weight was decreased dose dependently in rats treated neonatally with DES, but only the lowest dose group (0.01 microg) showed evidence of mating (3 of 6) and normal fertility (3 litters). Animals treated neonatally with OP or Bis-A had normal or increased (Bis-A) testis weights and exhibited reasonably normal mating/fertility. Animals fed a soy-free diet had significantly larger testes than controls fed a soy-containing diet, and this difference was confirmed in a much larger study of more than 24 litters, which also showed a significant decrease in plasma FSH levels and a significant increase in body weight in the males kept on a soy-free diet. Neonatal treatment with genistein did not alter adult testis weight, and although most males exhibited normal mating and fertility, a minority did not mate or were infertile. It is concluded that 1) neonatal exposure of rats to low levels of estrogens can advance the first wave of spermatogenesis at puberty, although it is unclear whether this is due to direct effects of the estrogen or to associated elevation of FSH levels; 2) the effect of high doses of OP and Bis-A on these processes is essentially benign; and 3) the presence or absence of soy or genistein in the diet has significant short-term (pubertal spermatogenesis) and long-term (body weight, testis size, FSH levels, and possibly mating) effects on males. PMID- 11014248 TI - 17beta-estradiol, but not raloxifene, decreases thrombomodulin in the antithrombotic protein C pathway. AB - Raloxifene is a nonsteroidal selective estrogen receptor modulator (SERM) that mimics the effects of estrogen on some plasma lipids and may have direct effects on the vascular wall. The objective of this study was to determine the effects of 17beta-estradiol, raloxifene, and LY139,478 (a related benzothiophene SERM) on the anticoagulant protein C pathway. In human vascular endothelial cells activated with interleukin-1 (IL-1), we demonstrated decreased thrombomodulin dependent protein C activation. 17beta-estradiol reduced the anticoagulant properties of both unstimulated and IL-1-activated endothelial cells by decreasing thrombomodulin expression. In contrast, raloxifene and LY139,478 enhanced the anticoagulant properties of both unstimulated and IL-1-activated endothelial cells through upregulation of thrombomodulin. Regulation of the protein C pathway via thrombomodulin on vascular endothelium may be a novel mechanism by which SERMs could potentially confer cardioprotective effects and reduce the thrombotic risk associated with HRT in compromised patients. PMID- 11014249 TI - Estrogen agonist and antagonist action on the human estrogen receptor in Drosophila. AB - The estrogen receptor (ER) regulates the expression of genes involved in the growth, proliferation and differentiation of skeletal, cardiovascular, neural and reproductive tissues. A basic scheme for the mechanism for ER action has been developed, but precise details on the interactions between ER and the cellular signaling and transcription machinery required for receptor-mediated regulation of specific target genes are still lacking. We have developed a genetic approach to explore the functional interactions of ER. In this work, we describe the development of an estrogen responsive system in the fruit fly, Drosophila melanogaster. Transgenic flies carrying the human ER alpha and an estrogen responsive green fluorescent protein (GFP) reporter gene were constructed. In vivo expression of the GFP reporter gene was observed when larvae were grown on a food source containing steroidal or nonsteroidal estrogens. The induction of the reporter gene by estrogens was blocked upon treatment with tamoxifen, an estrogen antagonist. However, we failed to recapitulate ligand-independent activation of the receptor in vivo or in cultured Drosophila cells. An estrogen responsive Drosophila system could be used to identify and characterize the complex functional interactions between ER and the other components of the cellular transcriptional apparatus. PMID- 11014251 TI - The evolving role of sonography in evaluating solid breast masses. AB - Many investigators have attempted to use ultrasound imaging to differentiate benign from malignant solid breast masses. Studies have evaluated several generations of gray-scale imaging, Doppler, color Doppler, and power Doppler imaging, and several unconventional ultrasound techniques. Although various individual studies have shown promise, ultrasound criteria for avoiding biopsy of solid lesions have not been widely adopted. Considerable observer variability also remains an important obstacle. This article reviews the results, strengths, and weaknesses of some of the key studies addressing this issue. Fundamental criteria for a successful ultrasound model are also specified. PMID- 11014252 TI - Use of Doppler ultrasound in the evaluation of breast carcinoma. AB - Ultrasound is an imaging modality commonly used to evaluate breast lesions in hopes to distinguish benign from malignant solid masses. Angiogenesis, defined as the emergence of new vessels to further the growth of tumor, has stimulated interest in the potential uses of Doppler ultrasound in patients with breast cancer. This article describes different forms of Doppler ultrasound, including color Doppler (CD), power Doppler (PD), and spectral Doppler (SD), as well as 3 dimensional (3D) ultrasound and ultrasound contrast media. We review the role of Doppler ultrasound in distinguishing benign from malignant solid breast masses. We also discuss the role of ultrasound in predicting tumor grade, histology, node status, and lymphatic vascular invasion, and in monitoring breast cancer treatment. PMID- 11014250 TI - Is the early increase in leptinemia one of the anorectic signals induced by an essential amino acid-deficient diet in the rat? AB - Rats start decreasing their food intake as early as 70 min after the first ingestion of a food deficient in threonine. A decrease of the limiting essential amino acid (EAA) in the plasma was proposed to be the first anorectic signal. Because many hormones regulate feeding behavior, we studied the effect of a meal (46 kJ) that was either devoid of threonine or was corrected for the deficiency, on plasma leptin, insulin and glucagon levels using a radio-immunoassay, at 0 to 180 min after the meal. One hour after ingestion of the threonine-devoid meal, a larger increase in insulinemia (22+/-1 vs. 15+/-1 microU/ml) and leptinemia (7.8+/-0.5 vs. 4.4+/-0.6 ng/ml; p<0.001) was observed than after ingestion of the corrected meal. The area under the curve of the threonine-devoid meal group was 3 and 1.34 fold larger than for the corrected meal group for insulin and leptin respectively. Glucagonemia was not different between the two groups. We propose that the rise in leptinemia, perhaps in synergy with rise in plasma insulin, might serve as one early signal to brain structures, participating in the anorectic mechanism following ingestion of an EAA-deficient diet. PMID- 11014253 TI - Texture analysis of breast tumors on sonograms. AB - We performed a feasibility study to determine if the texture features extracted from sonograms can be used to predict malignant or benign breast pathology by the proposed artificial neural network and to compare the diagnostic results with the radiologists' results. A total of 1,020 images (4 different rectangular regions from the 2 orthogonal imaging planes of each tumor) from 255 patients were used as samples. When a sonogram was performed, 1 physician identified the region of interest in the sonogram; then, a neural network model, using 24 autocorrelation texture features, classified the tumor as benign or malignant. Three radiologists who were unfamiliar with the samples also classified these images. The receiver operating characteristic (ROC) area index for the proposed neural network system is 0.9840 +/- 0.0072. The neural network identified 35 of 36 malignancies and 211 of 219 benign tumors using all 4 regions of interest. The radiologists, on average, identified 19 of 36 malignancies, with 12 tumors called indeterminate and 4 tumors called benign. We conclude that benign and malignant breast tumors can be distinguished using interpixel correlation in digital ultrasonic images. PMID- 11014254 TI - The role of ultrasound in the diagnosis of breast cancer. AB - Ultrasonography (US) of the breast is not applied to its optimal capacity if it is restricted to the differentiation between cystic and solid lesions and evaluation of dense breasts. It can play a major role in the detection of breast cancer; not only because of detection of mammographically occult lesions but also because of more accurate identification of lesion characteristics suspicious of malignancy. Its yield is highest among women younger than 50 years. However, one should keep in mind that the profit obtained by US shows not only the diagnostic accuracy of US but also that of mammography. Further improvement can be achieved by refinement in US diagnosis, especially with respect to the US characteristics of diffusely growing cancers. PMID- 11014255 TI - Clinically and mammographically occult breast lesions: detection and classification with high-resolution sonography. AB - With recent significant advances in ultrasound technology, the potential of high resolution sonography to improve the sensitivity of cancer diagnosis in women with dense breasts has become a matter of interest for breast imagers. To determine how often physician-performed high-resolution sonography can detect nonpalpable breast cancers that are not revealed by mammography, 8,970 women with breast density grades 2 through 4 underwent high-resolution sonography as an adjunct to mammography. All sonographically detected, clinically and mammographically occult breast lesions that were not simple cysts were prospectively classified into benign, indeterminate, or malignant categories. Diagnoses were confirmed by ultrasound-guided fine-needle aspiration, core-needle biopsy, or surgical biopsy. In 8,103 women with normal findings at mammography and physical examination, 32 cancers and 330 benign lesions were detected in 273 patients with sonography only. Eight additional cancers were found in 867 patients with a malignant (n = 5) or a benign (n = 3) palpable or mammographically detected index lesion. The overall prevalence of cancers detected with screening sonography was 0.41%, and the proportion of sonographically detected cancers to the total number of nonpalpable cancers was 22%. The mean size of invasive cancers detected only by sonography was 9.1 mm, and was not statistically different from the mean size of invasive cancers detected by mammography. The sensitivity of prospective sonographic classification for malignancy was 100%, and the specificity was 31%. In conclusion, the use of high-resolution sonography as an adjunct to mammography in women with dense breasts may lead to detection of a significant number of otherwise occult cancers that are no different in size from nonpalpable mammographically detected cancers. Prospective classification of these lesions based on sonographic characteristics resulted in an acceptable benign-to malignant biopsy rate of 6.3:1. PMID- 11014256 TI - Insecticidal rocaglamide derivatives from Aglaia spectabilis (Meliaceae). AB - Bark of Aglaia spectabilis collected on the island of Phu Quoc (Vietnam) yielded insecticidal cyclopentatetrahydrobenzofurans of the rocaglamide type including four new natural products. Structure elucidation of the new compounds is described. All rocaglamide derivatives isolated exhibited strong insecticidal activity towards neonate larvae of the polyphagous pest insect Spodoptera littoralis when incorporated into an artificial diet. LC50 values varied from 0.8 to 80 ppm. The most active compounds isolated, methylrocaglate and C-3' hydroxylmethylrocaglate, were similar with regard to their insecticidal activity to the well-known natural insecticide azadirachtin. PMID- 11014257 TI - A geranylated chalcone with 5alpha-reductase inhibitory properties from Artocarpus incisus. AB - A geranylated chalcone was isolated from leaves of Artocarpus incisus and it showed potent 5alpha-reductase inhibitory activity. PMID- 11014258 TI - Saikosaponins from roots of Bupleurum gibraltaricum and Bupleurum spinosum. AB - Two new saikosaponins have been identified in the butanolic fraction of the ethanol extract of the roots of Bupleurum spinosum: 3beta,16alpha,23,28 tetrahydroxyoleana-11,13(18)-dien- 30-oic acid 3-O-beta-D-glucopyranosyl-(1 --> 2)-beta-D-glucopyranosyl-(1 --> 3)-beta-D-fucopyranoside, and 3beta,16alpha,23,28,30-pentahydroxyoleana-11,13(18)-diene 3-O-beta-D glucopyranosyl-(1 --> 2)-beta-D-glucopyranosyl-(1 --> 3)-beta-D-fucopyranoside. 3Beta,16beta,23-trihydroxy-13,28-epoxyolean-11-ene 3-O-beta-D-glucopyranosyl-(1 - > 2)-beta-D-glucopyranosyl-(1 --> 3)-beta-D-fucopyranoside was also isolated, and this structure agreed with the one proposed for bupleuroside I, but their spectroscopic data have not been described until now. From the same fraction of the roots of Bupleurum gibraltaricum, the known compound buddlejasaponin IV has been isolated as the predominant component (90%). Structures were elucidated using spectroscopic analysis, specially 2D-NMR experiments. PMID- 11014259 TI - Volatile constituents of wood-rotting basidiomycetes. AB - Phytochemical investigation of the hydrodistillation products of the basidiomycetes Fomitopsis pinicola, Piptoporus betulinus, Gloeophyllum odoratum and Trametes suaveolens led to the identification of numerous mono- and sesquiterpenes as well as many aliphatic alcohols, aldehydes and ketones and some aromatic compounds. In addition, some diterpenes were identified as constituents of Fomitopsis pinicola. The absolute configuration of some terpenes was determined PMID- 11014261 TI - Constituents of the fungi Daedalea quercina and Daedaleopsis confragosa var. tricolor. AB - Phytochemical examination of solvent extracts of the wood-rotting fungi Daedalea quercina and Daedaleopsis confragosa var. tricolor led to the isolation of five new triterpene derivatives and some known fungal constituents. All structures were identified by one- and two-dimensional NMR spectroscopy and mass spectrometry. From Daedalea quercina, the new natural products 16-O acetylpolyporenic acid C, 16alpha-acetoxy-24-methylene-3-oxolanost-8-en-21-oic acid, (+)-24-methylene-3,23-dioxolanost-8-en-26-oic acid, (+)-3beta,12beta dihydroxy-24-methyl-23-oxolanost-8-en-26-oi c acid and 12beta,23-epoxy-3alpha,23 dihydroxy-24-methyllanost-8- en-26-oic acid could be isolated. From Daedaleopsis confragosa var. tricolor, the compounds 3alpha-carboxyacetoxyquercinic acid, 3alpha-carboxyacetoxy-24-methylene-23-oxolanost-8-en-2 6-oic acid and 5alpha,8alpha-epidioxyergosta-6,22-dien-3beta-ol were identified. These are the first described triterpene derivatives isolated from this fungus. PMID- 11014260 TI - Triterpene saponins and iridoid glucosides from galium rivale. AB - Three new glycosides of the oleanene-type triterpenes, rivalosides C-E (1-3), along with three known triterpene saponins, momordin IIb (4) and rivalosides A-B (5-6), and five known iridoid glucosides: monotropein, scandoside, deacetylasperulosidic acid, geniposidic acid and asperulosidic acid, were isolated from aerial parts of Galium rivale. The structures of the new compounds were elucidated by spectral methods and chemical means as 2alpha-acetoxy-3alpha, 19alpha-dihydroxy-olean-12-en-28-oic acid 28-O-beta-D-glucopyranosyl-(1--> 6) beta-D-glucopyranoside, 2alpha,3alpha, 19alpha-trihydroxy-olean- 12-en-28-oic acid 28-O-beta-D-glucopyranosyl-(1 --> 6)-beta-D-glucopyranoside and 3-O-beta-D glucuronopyranosyl-24-hydroxy-olean-12-en-28-oic acid 28-O-beta-D glucopyranoside, for rivalosides C-E, respectively. The taxonomic significance of the rivalosides in G. rivale was discussed. PMID- 11014262 TI - Sesquiterpenoids and phenolics from Crepis mollis. AB - From the roots of Crepis mollis, one new and two known guaianolides were isolated together with eight known guaianolide glycosides, one known germacranolide glycoside and two known phenylpropanoids. The structure and relative configuration of the new compound were established as 9alpha-hydroxy-4beta, 15, 11beta, 13-tetrahydro-dehydrozaluzanin C by spectral methods. PMID- 11014263 TI - Diterpenoids from the roots of Croton macrostachys. AB - Three novel diterpenoids have been isolated from the roots of Croton macrostachys. The structure and stereochemistry of the compounds have been unambiguously settled as neoclerodan-5,10-en-19,6beta;20,12-diolide, 3alpha, 19 dihydroxytrachylobane, and 3alpha,18,19-trihydroxytrachylobane from detailed spectroscopic evidence. PMID- 11014264 TI - Labdane diterpenes from Otostegia fruticosa. AB - The new labdane diterpenes otostegin A (2), otostegin B (6) and 15-epi-otostegin B (7) were isolated from the aerial parts of Otostegia. fruticosa, besides the previously known labdanes preleoheterin (1), leoheterin (3), leopersin C and 15 epi-leopersin C (4, 5), ballonigrin (9) and vulgarol (11), along with the iridoid glucoside 8-O-acetylharpagide (10). The structure elucidation of all the isolated compounds was based on their spectral data and chemical derivatization. PMID- 11014265 TI - Highly oxidized cuparene-type sesquiterpenes from a mycelial culture of Flammulina velutipes. AB - Cuparene-type sesquiterpenes were isolated from a culture broth of Flammulina velutipes (Curt.:Fr.) Sing. Using spectroscopic methods (HR-MS, 1H and 13C NMR, and 2D NMR, spectroscopy), their structures were determined to be 2,3,4,5 tetrahydro-2,7-dihydroxy-5,8,10,10-tetramethyl-2,5-methano-1- benzoxepin and 5 methyl-2-(3-oxo-1,2,2-trimethylcyclopentyl)benzoquinone. Both showed antimicrobial activity against Cladosporium herharum and Bacillus subtilis. PMID- 11014266 TI - Minor sulfated saikosaponins from the aerial parts of Bupleurum rigidum L. AB - Five new sulfated saikosaponins (Sandrosaponins II-VI) were isolated as minor components from the aerial parts of Bupleurum rigidum L. Their structures have been established by 1D and 2D-NMR techniques, FABMS, and chemical methods. , PMID- 11014267 TI - Sesquiterpenoids from roots of Taraxacum laevigatum and Taraxacum disseminatum. AB - Chromatographic separation of ethanolic root extracts of Taraxacum laevigatum and Taraxacum disseminatum afforded a total of eight germacrane- and eudesmane-type sesquiterpenoids. including new compounds, 1beta,3beta,6alpha-trihydroxy-4alpha( 15)-dihydrocostic acid methyl ester and its 1-O-beta-glucopyranoside. Their structures were established by spectroscopic analyses. In addition, the structure of 4alpha(15), 11beta(13)-tetrahydroridentin B-1-O-beta-glucopyranoside was elucidated by extensive NMR studies. PMID- 11014269 TI - Tirucallane triterpenoids from Dysoxylum hainanense. AB - Four tirucallane derivatives, 3beta,22S-dihydroxy-tirucalla-7,24-dien-23-one, 22,23-epoxy-tirucalla-7-ene-3beta,24,25-triol, 3beta,25-dihydroxy-tirucalla-7,23 diene, 23,26-dihydroxy-tirucalla-7,24-dien-3-one, together with two known triterpenoids, 24,25-epoxy-3beta,23-dihydroxy-7-tirucallene, tirucalla-7,24-diene 3beta,23-diol, were isolated from Dysoxylum hainanense. Their structures were elucidated on the basis of spectroscopic evidence. PMID- 11014268 TI - Saponins from Lonicera bournei. AB - The lupane-triterpene glycosides, bourneioside A and bourneioside B, and two known saponins were isolated from Lonicera bournei Hemsl. The structures of bourneioside A and B were elucidated as 3-O-beta-D-glucopyranosyl-23-hydroxy-lup 20(29)-en-28-oic acid-28-O-beta-D-glucopyranosyl ester and 3-O-beta-D glucopyranosyl-23-hydroxy-lup-20(29)-en-28-oic acid-28-O-[beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranosyl] ester, respectively, on the basis of spectral data and chemical evidence. PMID- 11014270 TI - Iridoids from Caryopteris x clandonensis. AB - In continuation of our phytochemical studies on Caryopteris x clandonensis (Lamiaceae), three further iridoids were isolated from the methanolic extract of the stems. Their structures were established by 1D and 2D NMR and MS analysis as a C-6 epimer of 8-O-acetylharpagide (6-epi-8-O-acetylharpagide), a derivative of harpagide which contained the unusual feature of a 3',4' seco-glycopyranosyl moiety (clandonoside II) and a methyl cetal of 8-O-acetylharpagide aglucone hydrate named clandonensine. PMID- 11014271 TI - Sesquiterpene lactone and friedelane derivative from drypetes molunduana. AB - A new sesquiterpene lactone, drypemolundein A and a new friedelane derivative, drypemolundein B, along with seven known compounds have been isolated from the whole stems of Drypetes molunduana Pax and Hoffm. Their structures were established on the basis of one- and two-dimensional NMR, homo- and hetero nuclear spectroscopic evidence. PMID- 11014272 TI - Triterpenoid saponins from Ardisia mamillata. AB - Two saponins were isolated from the roots of Ardisia mamillata HANCE. Their structures were established on the basis of MALDI-TOFMS, 1H, 13C NMR and 2D NMR (COSY, HOHAHA, HETCOR, HMBC and ROESY) spectra, and on chemical evidence, to be ardisimamilloside A, 3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl (1 --> 4)-[beta-D-glucopyranosyl-(1 --> 2)]-alpha-L-arabinopyranosyl]-3beta, 16alpha,28alpha-trihydroxy-13beta,28-epoxy-oleanan+ ++-30-al; and ardisimamilloside B, 3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl (1 --> 4)-[beta-D-glucopyranosyl-( 1 --> 2)]-alpha-L-arabinopyranosyl]-beta3 hydroxy-13beta,28- epoxy-oleanan-16-oxo-30-al. PMID- 11014273 TI - Two oleanane triterpenoids from gordonia ceylanica and their conversions to taraxarane triterpenoids. AB - Chemical investigation of the hot hexane extract of the stem bark of Gordonia ceylanica afforded two new oleanane triterpenoids, 3beta-acetoxy-11alpha, 13beta dihydroxyolean-12-one and 3beta,11alpha-diacetoxy-13beta-hydroxyolean-12-one (2) The attempted acid hydrolysis of these two compounds resulted the dehydration and subsequent methyl group migration to afford the taraxarane triterpenoids 3beta,11alpha-dihydroxytaraxer-14-en-12-one (4) and 3beta-hydroxy-11alpha acetoxytaraxer-14-en-12-one (5), respectively. These taraxaranes have not been previously reported. PMID- 11014274 TI - Taxane diterpenoids from Taxus yunnanensis and Taxus cuspidata. AB - Chemical examination of the seeds of the Chinese yew, Taxus runnanensis Cheng et L. K. Fu and the Japanese yew, Taxus cuspidata Sieb et Zucc, resulted in the isolation of four taxane diterpenoids. The structures of these taxoids were established as (12alpha)-2alpha-acetoxy-5alpha,9alpha, 10beta-trihydroxy-3,11 cyclotax-4(20)-en-13-one; 2alpha,7beta,13alpha-triacetoxy-5alpha, 9alpha dihydroxy-2(3-->20)abeotaxa-4(20),11-dien-10-one; 9alpha,10beta-diacetoxy-5alpha cinnamoyloxytaxa- 4(20),11-dien-13alpha-ol and the known 2alpha,7beta,9alpha,10beta,13-pentaacetoxytax a-4(20),12-diene-5alpha,11beta-diol on the basis of spectral analysis. PMID- 11014275 TI - Euphane triterpenoid and lipid constituents from Butea monosperma. AB - Besides stigmasterol, stigmasterol-betaD-glucopyranoside and nonacosanoic acid, two new compounds isolated from the stems of Butea monosperma have been characterised as 3alpha-hydroxyeuph-25-ene and 2,14-dihydroxy-11,12-dimethyl-8 oxo-octadec-11-enylcyclohexane+ ++ by spectral data and chemical studies. PMID- 11014276 TI - Neutral taxoids from Taxus cuspidata as modulators of multidrug-resistant tumor cells. AB - Two taxoids, taxinine NN-7 (1) and 3,11-cyclotaxinine NN-2 (2), were isolated from the neutral fraction of the EtOAc extract of a mixture of needles and young stems of Taxus cuspidata. The structures were determined by spectroscopic analysis. Both compounds showed some activity as modulators of multidrug resistant tumor cells. PMID- 11014277 TI - Diterpene glucosides from Pieris formosa. AB - Five diterpene glucosides, pierisformosides B-F were isolated from Pieris formosa D. Don (Ericaceae). Their structures were elucidated on the basis of spectral analysis. including 1H-1H COSY, 13C-1H COSY, HMBC and NOESY experiments. PMID- 11014278 TI - Triterpenoid saponins from Fagonia cretica. AB - Four new triterpenoid saponins were isolated and identified from the aerial parts of Fagonia cretica. They were characterized as 3-O-[beta-D-glucopyranosyl (1-->2) alpha-L-arabinopyranosyl] hederagenin 28-O-beta-D-glucopyranosyl ester, 3-O-[beta D-glucopyranosyl (1-->2)-alpha-L-arabinopyranosyl] oleanolic acid 28-O-[beta-D glucopyranosyl (1-->6)-beta-D-glucopyranosyl] ester, 3-O-[beta-D-glucopyranosyl (1-->2)-alpha-L-arabinopyranosyl] 27-hydroxy oleanolic acid 28-O-[beta-D glucopyranosyl (1-->6)-beta-D-glucopyranosyl] ester and 3beta-O-[beta-D glucopyranosyl (1-->2)-alpha-L-arabinopyranosyl] olean-12-en-27-al-28-oic acid 28 O-[beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranosyl] ester. The structures of the saponins were assigned by spectral analyses (FABMS, 1H, 13C NMR, 1H-1H COSY, TOCSY, HMQC and HMBC spectra) and NOE experiments. To the best of our knowledge the genin 3beta hydroxy olean-12-en-27-al-28-oic acid is new. PMID- 11014279 TI - Triterpenoidal constituents from Eucalyptus camaldulensis var. obtusa leaves. AB - An investigation on the constituents of the fresh, uncrushed leaves of Ecalyptus camaldulensis var. obtusa has led to the isolation of the triterpenoid amirinic acid and four known triterpenoids ursolic acid lactone, betulinic acid, oleanolic acid and ursolic acid. Amirinic acid transformed into amirolide in deuterated chloroform at room temperature. The new products were characterized by exhaustive spectroscopic studies as 2alpha,3beta,7beta-trihydroxy-11alpha-methoxyurs -12-en 28-oic acid and 2alpha,3beta,7beta-trihydroxyurs-11-en-28,13beta -olide. PMID- 11014280 TI - Prieurianoside, a protolimonoid glucoside from the leaves of Trichilia prieuriana. AB - The ubiquitous glycolipid 1,2-dilinolenoyl-3-galactopyranosylglycerol and a new protolimonoid glucoside, named prieurianoside, were isolated from the leaves of Trichilia prieuriana. The structure of the latter was established, by spectroscopic techniques, as 12beta,21-diacetoxy-29-beta-D-glucopyranosyloxy 23zeta -hydroxytirucalla-7,24-dien-3-one. PMID- 11014281 TI - A stilbene and dihydrochalcones with radical scavenging activities from Loiseleuria procumbens. AB - A new dihydrochalcone, 6''-acetylphloridzosid, was isolated from the whole plant of Loiseleuria procumbens (L.) Desv. and identified as 2'-O-(6''-O acetylglucopyranosyl)-4,4',6'-trihydroxydihydrochal cone by spectroscopic methods. In addition, one stilbene and three other dihydrochalcones were identified as (E)-piceid, phloretin (2',4,4',6'-tetrahydroxydihydrochalcone), phloridzosid (2'-O-glucopyranosyl-4,4',6'-trihydroxydihydrochalcone) and asebotin (2'-O-glucopyranosyl-4'-methoxy-4,6'-dihydroxydihydrochalcone), respectively. Some of these compounds showed scavenging properties towards the 2,2-diphenyl-1 picrylhydrazyl radical and antioxidant properties in a test with lysozyme. PMID- 11014282 TI - Stilbene dimers from the lianas of Gnetum hainanense. AB - Five stilbene dimers, gnetuhainins F-J, were isolated together with gnetulin, rhapontigenin, isorhapontigenin and gnetol from the lianas of Gnetum hainanense C. Y. Cheng. Their structures and stereochemistry have been established on the basis of spectral evidence, especially 2D NMR spectroscopic techniques. PMID- 11014283 TI - Benzophenone glycosides from Gnidia involucrata. AB - Six compounds have been isolated from the methanol extract of the aerial parts of Gnidia involucrata (Thymelaeaceae). They were identified as 2,3,4',5,6 pentahydroxybenzophenone-4-C-glucoside and 2,4',6-trihydroxy-4 methoxybenzophenone-2-O-glucoside, together with mangiferin, kaempferol-3-O glucoside, yuankanin and manniflavanone by chemical and spectroscopic means. The structures of three additional C-glycosyl flavones--vitexin, isovitexin and isoorientin--were determined on-line by LC/UV/APCI-MSn analysis of the crude extract. PMID- 11014284 TI - Minor phenolics from Crinum bulbispermum bulbs. AB - From the bulbs of Crinum bulbispermum Milne, four new minor compounds were isolated viz. 4-hydroxy-2',4'-dimethoxydihydrochalcone (1), 4,5-methylenedioxy-4' hydroxy-2-aldehyde[1,1'-biphenyl] (4), hippacine (6), and 4'-hydroxy-7 methoxyflavan-3-ol (7). In addition, four known compounds were isolated and identified as 2(S),3',4'-dihydroxy-7-methoxy flavan (2), isolarrien (3), isoliquiritigenin (5) and liquiritigenin (8). The structures of the isolated compounds were established by spectral evidence. PMID- 11014285 TI - Spectral comparisons of coniferyl and cinnamyl alcohol epoxide derivatives with a purported cis-epoxyconiferyl alcohol isolate. AB - The reported isolation of cis-epoxyconiferyl alcohol must be incorrect, based upon comparison of the reported Nuclear Magnetic Resonance (NMR) spectral data for the isolate with those for synthesized coniferyl and cinnamyl alcohol epoxide derivatives. Attempts to prepare cis- and trans-coniferyl alcohols were unsuccessful, although their acetate derivatives could be synthesized. The NMR spectral data for a synthetic sample of pinoresinol were in excellent agreement with those for the purported isolate. PMID- 11014286 TI - Dihydroflavonols from Lannea coromandelica. AB - The dihydroflavonols, (2R,3S)-(+)-3',5-dihydroxy-4',7-dimethoxydihydroflavonol and (2R,3R)-(+)-4',5,7-trimethoxydihydroflavonol were isolated from the stem bark of Lannea coromandelica, along with the known (2R,3R)-(+)-4',7-di-O methyldihydroquercetin, (2R,3R)-(+)-4',7-di-O-methyldihydrokaempferol and (2R,3R) (+)-4'-O-methyldihydroquercetin. All five compounds were isolated for the first time from the genus Lannea; furthermore, (2R,3S)-(+)-3',5-dihydroxy-4',7 dimethoxydihydroflavonol, was a rare cis-type isomer. The structures of all compounds were elucidated by spectroscopic methods including 2D NMR and CD analysis. PMID- 11014287 TI - Lignans from Mosla scabra. AB - Two new cyclobutane-type lignans, named moslolignans A and B, together with two known ones, andamanicin and magnosalin, were isolated from the whole plant of Mosla scabra. Their structures were established as 1beta*,2beta*,3alpha*,4alpha* 1,2-dimethyl-3-(3- methoxy-4,5-methylene-dioxyphenyl)-4-(2,4,5-trimethoxyphenyl) cycl obutane and 1beta*,2beta*,3alpha*,4alpha*-1,2-dimethyl-3-(2, 5-dimethoxy-3,4 methylenedioxyphenyl)-4-(2,4,5-trimethoxyphenyl)-cyclobu tane by spectroscopic methods. This is the first report of naturally-occurring cyclobutane-type lignans with asymmetrical substitutions. PMID- 11014288 TI - Acylated delphinidin 3-rutinoside-5-glucosides in the flowers of Petunia reitzii. AB - Two acylated anthocyanins were isolated from selected individuals of Petunia reitzii, and identified to be delphinidin 3-O-[6-O-(4-O-(4-O-(6-O-(trans caffeoyl)-beta-D-glucopyranosyl)-tr ans-p-coumaroyl)-alpha-L-rhamnopyranosyl) beta-D-glucopyranoside]- 5-O-[beta-D-glucopyranoside] and delphinidin 3-O-[6-O-(4 O-(4-O-(beta-D-glucopyranosyl)-trans-p-coumaroyl)-alph a-L-rhamnopyranosyl)-beta D-glucopyranoside]-5-O-[beta-D-glucopyranoside ]. Nine known anthocyanins were also identified. PMID- 11014289 TI - Anthocyanins in callus induced from purple storage root of Ipomoea batatas L. AB - Two anthocyanins were isolated from the highly pigmented callus derived from the storage root of purple sweet potato (Ipomoea batatas L.) cultivar 'Ayamurasaki'. One was identified as cyanidin 3-O-sophoroside-5-O-glucoside, and the other as cyanidin 3-O-(2-O-(6-O-(E)-p-coumaroyl-beta-D-glucopyranosyl)-beta-D-glucop yranoside)-5-O-beta-D-glucopyranoside, by chemical and spectroscopic analysis. PMID- 11014290 TI - Phenylpropanoid glycosides from Scrophularia ningpoensis. AB - Three phenylpropanoid glycosides named ningposides A (3-O-acetyl-2-O-feruloyl alpha-L-rhamnopyranose), B (4-O-acetyl-2-O-feruloyl-alpha-L-rhamnopyranose) and C (3-O-acetyl-2-O-p-hydroxycinnamoyl-alpha-L-rhamnopyranose) along with the known compounds sibirioside A, cistanoside D, angoroside C, acteoside, decaffeoylacteoside and cistanoside F were obtained from the roots of Scrophularia ningpoensis. PMID- 11014291 TI - Tropane alkaloids from the leaves and stem bark of Erythroxylon alaternifolium and Erythroxylon rotundifolium. AB - A novel 3alpha,6beta,7beta-triol tropane alkaloid esterified by two benzoyl residues was isolated from the leaves of the endemic cuban species, Erythroxylon alaternifolium. Another novel 3alpha,6alpha,7beta-triol tropane alkaloid esterified by trimethoxycinnamoyl and trimethoxybenzoyl residues was isolated from the leaves and stem bark of a second endemic cuban species, Erythroxylon rotundifolium. Their structures were elucidated as 3alpha,7beta-dibenzoyloxy 6beta-hydroxy-tropane and 3alpha-(3,4,5-trimethoxycinnamoyloxy)-7beta-(3,4,5- trimethoxybenzoyloxy)-6alpha-hydroxy-tropane by spectroscopic methods including 2D-NMR techniques. PMID- 11014292 TI - Structure revision of isoline (ruwenine), bisline and isolinecic acid. AB - X-ray crystallography of bisline, and the chemical interconversion of bisline and isoline (ruwenine), revealed that the structures previously assigned to these alkaloids required revision; as did that of isolinecic acid. PMID- 11014293 TI - Caffeine: also a fungal metabolite. AB - Caffeine has been found to occur as a fungal metabolite and to be the principal alkaloid in sclerotia of Claviceps sorghicola, a Japanese ergot pathogen of Sorghum spp. PMID- 11014294 TI - Indole alkaloids from a culture of the fungus Aporpium caryae. AB - In a screening for antifungal metabolites, two indole compounds of mixed biogenesis, 1H-indole-3-carboxylic acid, 1-(1,1-dimethyl-2-propenyl) methyl ester and 1H-indole-3-carboxylic acid, 1-(2,3-dihydroxy-1,1-dimethylpropyl) methyl ester were isolated from a culture of the basidiomycete Aporpiums caryae. The structural elucidation of these compounds was accomplished by spectroscopic methods. PMID- 11014295 TI - Alkaloids from Crinum macowanii. AB - Eleven alkaloids have been isolated from fresh bulbs of Crinum macowanii (Amaryllidaceae). Macowine is reported here for the first time. The structure and stereochemistry of this new alkaloid as well as of the known ones were determined by physical and spectroscopic methods. PMID- 11014296 TI - Four steroidal alkaloids from the leaves of Buxus sempervirens. AB - Four new steroidal alkaloids, N20-formylbuxaminol E [(20S)-16alpha-hydroxy-20 (formylamino)-3beta-(dimethylamino)-9,10 -seco-buxa-9(11),10(19)-diene] (1), O16 syringylbuxaminol E [(20S)-16alpha-syringoyl-3beta-(dimethylamino)-20-(amino)-9, 10-seco-buxa-9(11),10(19)-diene] (2), N20-acetylbuxamine G [(20S)-20 (acetylamino)-3beta-(methylamino)-9,10-seco-buxa-9(11),1 0(19)-diene] (3) and N20 acetylbuxamine E [(20S)-20-(acetylamino)-3beta-(dimethylamino)-9,10-seco-buxa 9(11) ,10(19)-diene] (4) were isolated from the leaves of Buxus sempervirens. Their structures were determined mainly on the basis of 2D NMR studies. PMID- 11014297 TI - Alkaloids from Isopyrum thalictroides L. AB - Three new bisbenzylisoquinoline alkaloids, isopyruthaldine, isopythaldine and isothalmidine, containing new type of bridges were isolated from roots and rhizomes of Isopyrum thalictroides. The known isoquinoline alkaloids, reticuline, isocorydine. columbamine and palmatine, obtained from the aerial parts, are new for the genus Isopyrum. All structures were elucidated by spectroscopic analysis. PMID- 11014298 TI - Fatty acid amides from freshwater green alga Rhizoclonium hieroglyphicum. AB - Freshwater green algae Rhizoclonium hieroglyphicum growing in the Ural Mountains were examined for their fatty acid amides using capillary gas chromatography-mass spectrometry (GC-MS). Eight fatty acid amides were identified by GC-MS. (Z)-9 octadecenamide was found to be the major component (2.26%). PMID- 11014299 TI - Triterpenoids, essential oil and photo-oxidative 28 --> 13-lactonization of oleanolic acid from Lantana camara. AB - Two novel triterpenoids have been isolated from the roots of Lantana camara L.: 3beta,19alpha dihydroxy ursan-28-oic acid and 21,22beta-epoxy-3beta-hydroxy olean 12-en-28-oic acid in its methyl ester form. Its leaves have yielded an essential oil which is rich in sesquiterpenes. Oleanolic acid, which is thought to be a hepatoprotective compound, was isolated from L. camara roots and converted into its 28 --> 13beta lactone by a facile photo-oxidation reaction. PMID- 11014300 TI - Diarylheptanoids from Myrica arborea. AB - Investigations of the stem and root bark of Myrica arborea (Myricaceae) have yielded two novel diarylheptanoids, myricarborin and 11-O-beta-D xylopyranosylmyricanol along with the known myricanol and 5-O-beta-D glucopyranosylmyricanol. The structures of the novel compounds were determined by spectroscopic methods. PMID- 11014301 TI - Constituents of Chondria armata. AB - A novel long chain fatty ester, pentyl hentriacontanoate 1 and an orange red pigment, caulerpin 2 have been isolated and characterised from a red alga Chondria armata. The pigment caulerpin hitherto known to be a constituent of green algae of genus Caulerpa is being reported here for the first time from a red alga. PMID- 11014302 TI - C21 steroidal glycosides from Hemidesmus indicus. AB - Two novel pregnane glycosides, denicunine (1) and heminine (4), have been isolated from the dried stem of Hemidesmus indicus R.Br. (family: Asclepiadaceae). Chemical transformations and spectroscopic evidence viz: 1H and 13C NMR spectroscopy and FABMS are consistent with the structures calogenin 3-O-3 O-methyl-beta-D-fucopyranosyl-(1-->4)-O-beta-D-oleandropyranosi de and calogenin 3-O-beta-D-cymaropyranosyl-(1-->4)-O-beta-D-digitoxopyranoside+ ++, respectively. PMID- 11014303 TI - A 6-substituted-5,6-dihydro-2-pyrone from Cryptocarya strictifolia. AB - The structure and absolute configuration of an alpha-pyrone isolated from Cryptocarya strictifolia was elucidated as 6R-(4'R,6'R-dihydroxy-8'-phenyloct-1' enyl)-5,6-dihydro-2H-pyran-2 -one. Pinocembrin and lysicamine were also isolated. PMID- 11014304 TI - Fluorescence studies of ATP-diphosphohydrolase from Solanum tuberosum var. Desiree. AB - Chemical modification of potato apyrase suggests that tryptophan residues are close to the nucleotide binding site. Kd values (+/- Ca2+) for the complexes of apyrase with the non-hydrolysable phosphonate adenine nucleotide analogues, adenosine 5'-(beta,gamma-methylene) triphosphate and adenosine 5'-(alpha,beta methylene) diphosphate, were obtained from quenching of the intrinsic enzyme fluorescence. Other fluorescent nucleotide analogues (2'(3')-O-(2,4,6 trinitrophenyl) adenosine 5'-triphosphate, 2'(3')-O-(2,4,6-trinitrophenyl) adenosine 5'-diphosphate. 1,N6-ethenoadenosine triphosphate and 1,N6 ethenoadenosine diphosphate) were hydrolysed by apyrase in the presence of Ca2+, indicating binding to the active site. The dissociation constants for the binding of these analogues were calculated from both the decrease of the protein (tryptophan) fluorescence and enhancement of the nucleotide fluorescence. Using the sensitised acceptor (nucleotide analogue) fluorescence method, energy transfer was observed between enzyme tryptophans and ethene-derivatives. These results support the view that tryptophan residues are present in the nucleotide binding region of the protein, appropriately oriented to allow the energy transfer process to occur. PMID- 11014305 TI - Trauma and pressure explain the clinical presentation of the Graves' disease triad. PMID- 11014306 TI - Impaired response to hepatocyte growth factor in FRTL-5 rat thyroid cells expressing a functional hepatocyte growth factor receptor. AB - The FRTL-5 rat thyroid cell line is widely used for studies of thyrocyte function and growth. The objective of the present study was to investigate the effects of hepatocyte growth factor (HGF) on FRTL-5 cells. HGF has previously been known to be a potent regulator of thyrocyte growth and differentiation. Met, the receptor for HGF was expressed in FRTL-5 cells, as well as in primary cultures of porcine thyrocytes included in the study as control. On HGF stimulation Met was tyrosine phosphorylated in both porcine and FRTL-5 cells, indicating an activation of the receptor. Addition of HGF induced changes of cell shape, scattering and proliferation of the porcine thyrocytes, but not in the FRTL-5 cells; yet, a functional coupling of Met to the p85 subunit of the phosphatidylinositol-3' kinase (PI3'-K) in coprecipitation experiments, formation of focal adhesions detected in immunofluorescence staining with an antivinculin antibody, and induction of c-fos mRNA in Northern blot analysis was observed in FRTL-5 cells, showing a transduction of the HGF/Met signal. In summary, despite the expression of apparently functional Met, the FRTL-5 cells are unable to properly respond to HGF. PMID- 11014307 TI - Thyroid hormones influence serum leptin levels in patients with Graves' disease during suppression of beta-adrenergic receptors. AB - Leptin is a protein product of the ob gene, mainly produced by adipocytes. Leptin is thought to play an important role in the homeostasis of body weight by suppressing appetite and increasing energy consumption. The aim of this study was to investigate the possible effect of thyroid hormone on the regulation of the leptin system during suppression of beta-adrenergic receptors in Graves' patients. We studied 15 adult female patients with Graves' disease. Thyroid function, serum levels of leptin, and percent body fat (%BF) were examined at four different clinical conditions during therapy (A, untreated; B, beta adrenergic antagonist only [A, B; hyperthyroid], C, beta-adrenergic antagonist and antithyroid drug; D, antithyroid drug only [C, D; euthyroid]). The use of beta-adrenergic antagonist significantly reduced heart rate in spite of hyperthyroid state, indicating sufficient suppression of beta-adrenergic receptors. During treatment with beta-adrenergic antagonist, leptin percentage of body fat (%BF) ratio significantly decreased in euthyroid state compared to that in hyperthyroid state (from 38.7 +/- 21.3 to 18.1 +/- 19.3, p = 0.003). Moreover, there was a significantly positive correlation between delta leptin/%BF and delta free thyroxine (FT4) (r = 0.51, p = 0.008). Under a euthyroid state induced by antithyroid drug treatment, leptin/%BF did not change in spite of withdrawal of beta-adrenergic antagonist. Our data indicate that thyroid hormones could increase serum leptin level during suppression of beta-adrenergic receptors in Graves' patients. Our data also suggest that the beta-adrenergic action of thyroid hormones might be partly mediated by regulation of leptin. PMID- 11014308 TI - The somatostatin analogue octreotide modulates Iodothyronine deiodinase activity and pituitary neuromedin B. AB - Somatostatin inhibits growth hormone and thyrotropin (TSH) secretion. It also enhances the inhibitory effect of thyroid hormone (TH) on TSH by poorly understood mechanisms. We investigated the acute effect of the long-acting somatostatin analogue, octreotide (OCT), on anterior pituitary type 1 (D1) and 2 (D2) deiodinase activity, on liver D1, and on pituitary content of neuromedin B (NB), an autocrine inhibitor of TSH secretion, which is positively regulated by thyroid hormones. Euthyroid or hypothyroid rats were sacrificed at different times after a single subcutaneous injection of OCT (1 microg/kg body weight [BW]). D1 and D2 activities were measured by the release of 125I from 125I reverse triiodothyronine (rT3) under different assay conditions. NB, TSH, T3, and thyroxine (T4) were quantitated by radioimmunoassay (RIA). In euthyroid rats, liver and pituitary D1 activities were decreased (50%) 6 hours after OCT injection; pituitary D2 and NB remained unchanged. In hypothyroid rats, OCT increased near to the level of normal rats both pituitary D1 activity (but not liver) and NB content, at 24 hours and at 6 and 24 hours, respectively (p < 0.05). Pituitary D2, greatly increased by hypothyroidism, showed a small (25%) but significant reduction at 3 hours, persisting at 24 hours (p < 0.01), although it remained higher than that of euthyroid control. Serum thyroid hormones were not affected by OCT injection. The results show that octreotide acutely regulates pituitary deiodinases and NB content, both representing mechanisms that potentially can contribute to somatostatin and octreotide actions on pituitary growth hormone (GH) and TSH secretion and to modulate these cells sensitivity to thyroid hormone action. PMID- 11014309 TI - Sensitive thyroid-stimulating antibody assay in whole serum containing five percent polyethylene glycol using porcine thyroid cells. AB - Previously we reported that the amounts of cyclic adenosine monophosphate (cAMP) production by polyethylene glycol (PEG) 22.5% precipitated fraction (PF) (crude immunoglobulin G [IgG]) from thyroid-stimulating antibody (TSAb)-positive serum were higher than those by PEG 12.5% PF, and that PEG (5%) augmented affinity purified TSAb-IgG-stimulated cAMP production in porcine thyroid cells (PTC) assay. In the present work, we studied sensitive TSAb assay using whole sera in the presence of high PEG concentrations in PTC assay. cAMP produced by TSAb positive serum increased in proportion to serum amounts up to 0.05 mL, but gradually decreased in 0.075 mL. The maximal augmentative effect of PEG on TSAb positive serum (0.05 mL)-stimulated cAMP production was found in 5% PEG (final). Thus, TSAb assay using whole serum (0.05 mL) in the absence of 5% PEG (serum method) and sensitive TSAb (sTSAb) assay using whole serum (0.05 mL) in the presence of 5% PEG (serum plus 5% PEG method) were performed. The sensitive thyroid-stimulating antibody (sTSAb) activities of Graves' sera showed significantly higher (twofold to sevenfold) compared to simple TSAb activity with sera. sTSAb and TSAb activities were positive in 91% (29/32) and 47% (15/32) of untreated Graves' patients with hyperthyroidism. The sTSAb activities by serum plus 5% PEG method were higher than that by PEG 12.5% precipitated fraction (PF) from test serum (0.2 mL) in many Graves' sera, but lower than that by PEG 22.5% PF from test serum (0.2 mL). PEG (5%) did not augment cAMP produced by high TSH serum (127-210 mU/L) in thyroiditis chronica. sTSAb activity was negative in adenomatous goiter, subacute thyroiditis, and thyroid cancer. sTSAb activity was also negative in TSH stimulation blocking antibody (TSBAb)-positive hypothyroidism (during thyroxine [T4] treatment), but was positive in Graves' sera with coexistence serum of TSAb and TSBAb because of augmentative effect of 5% PEG on TSAb activity. This assay in whole serum (0.05 mL) containing 5% PEG is less sensitive than sensitive TSAb assay using PEG 22.5% PF from test serum (0.2 mL), but this method can be available clinically as routine TSAb assay using whole serum because of the technical simplicity. PMID- 11014310 TI - The effect of short-term low-dose perchlorate on various aspects of thyroid function. AB - Perchlorate (ClO4) salts are found in rocket fuel, fireworks, and fertilizer. Because of ground water contamination, ClO4 has recently been detected in large public water supplies in several states in the 4-18 microg/L (parts per billion [ppb]) range. The potential adverse effect of chronic low level ClO4 ingestion on thyroid function is of concern to the Environmental Protection Agency (EPA). The daily ingestion of ClO4 at these levels would be magnitudes below the therapeutic effect level of hundreds of milligrams of ClO4 used in treating hyperthyroidism. Studies were carried out in nine healthy male volunteers who had normal thyroid function and negative thyroid antibodies to determine whether the ingestion of 10 mg of ClO4 daily (approximately 300 times the estimated maximum amount of ClO4 consumed from the affected water supplies) would affect any aspect of thyroid function. They ingested 10 mg of ClO4 dissolved in a liter of spring water during waking hours for 14 days. Baseline serum thyrotropin (TSH), free thyroxine index (FTI), total triiodothyronine (TT3), 4-, 8-, and 24-hour thyroid 123I uptakes (RAIU), serum and 24-hour urine ClO4, 24-hour urine iodine, complete blood count (CBC), and chemistry profile were determined. All blood and urine tests were repeated on days 7 and 14 of ClO4 administration and thyroid RAIU on day 14 of ClO4 administration. All tests were repeated 14 days after ClO4 was discontinued. No effect of ClO4 on serum thyroid hormone or TSH concentrations, urinary iodine excretion, CBC, or blood chemistry was observed. Urine and serum ClO4 levels were appropriately elevated during the course of ClO4 ingestion in all subjects, demonstrating compliance. By day 14 of ClO4 administration, the 4-, 8-, and 24 hour thyroid RAIU values decreased in all nine subjects by a mean value of 38% from baseline and rebounded above baseline values by 25% at 14 days after ClO4 withdrawal (p < 0.01 analysis of variance (ANOVA) and Tukey). It is well known that the major effect of ClO4 on the thyroid is a decrease in the thyroid iodide trap by competitive inhibition of the sodium iodide symporter (NIS). The present study demonstrates the sensitivity of the thyroid iodide trap to ClO4 because a low dose of 10 mg daily significantly decreased the thyroid RAIU without affecting circulating thyroid hormone or TSH concentrations. It is possible, however, that the daily consumption of low levels of ClO4 in drinking water over a prolonged period of time could adversely affect thyroid function but no evidence of hypothyroidism was observed at 10 mg of ClO4 daily in this 2-week study. It is now of interest to determine a no effect level for ClO4 on the inhibition of the thyroid RAIU and to carry out a long-term ClO4 exposure study. PMID- 11014311 TI - Cardiovascular and atherogenic aspects of subclinical hypothyroidism. AB - Subclinical hypothyroidism (SH) is common, especially among elderly women. There is no clear evidence to date that SH causes clinical heart disease. However, mild thyroid gland failure, evidenced solely by elevation of the serum thyrotropin (TSH) concentration, may be associated with increased morbidity, particularly for cardiovascular disease, and subtly decreased myocardial contractility. In SH, both cardiac structures and function remain normal at rest, but impaired ventricular function as well as cardiovascular and respiratory adaptation to effort may become unmasked during exercise. These changes are reversible when euthyroidism is restored. Flow-mediated vasodilatation, a marker of endothelial function, is significantly impaired in SH, and decreased heart rate variability, a marker of autonomic activity, suggests hypofunctional abnormalities in the parasympathetic nervous system. SH does result in a small increase in low-density lipoprotein (LDL) cholesterol (C) and a decrease in high-density lipoprotein (HDL)-C, changes that enhance the risk for development of atherosclerosis and coronary artery disease (CAD). After coronary revascularization, a trend toward higher rates of chest pain, dissection, and reocclusion has been noted in SH subjects. Smoking may contribute to the high incidence of SH and may aggravate its metabolic effects. Subjects with SH with marked TSH elevation and high titers of thyroid autoantibodies are at higher risk of unnoticed progression to overt hypothyroidism. Especially women over 50 years with TSH levels greater than 10 mU/L and smoking habits have the highest risk for cardiovascular complications. The magnitude of the lipid changes and the subtle impairment of left ventricular function and cardiopulmonary exercise capacity in SH may justify use of hormone replacement. Early levothyroxine (LT4) treatment in SH may reduce the C level by an average of 8% and normalize all metabolic effects in smokers, nevertheless, in some patients, LT4 therapy may exacerbate angina pectoris or an underlying cardiac arrhythmia. Longitudinal follow-up to define the actual cardiovascular disease risk associated with SH is warranted. PMID- 11014312 TI - Complacency: the most dangerous enemy in the war against iodine deficiency. PMID- 11014313 TI - Elephantiasic pretibial myxedema: insight into and a hypothesis regarding the pathogenesis of the extrathyroidal manifestations of Graves' disease. AB - The basis for the extrathyroidal manifestations of Graves' ophthalmopathy (GO) and dermopathy are not well understood. We describe immunohistochemical studies on the skin of a patient with an extreme, elephantiasic form of Graves' dermopathy that developed after periods of prolonged standing with dependent edema. Excision of part of the lesion with subsequent skin grafting from a normal donor site resulted in recurrence of the disease at the original site as well as in development of disease at the donor site. A murine monoclonal antibody reacted with the thyrotropin receptor (TSHR) or a cross-reacting protein in fibroblast like cells in the patient's upper dermis and, surprisingly, with dermal cells from unaffected individuals. The patient's dermis containing lymphoid follicles comprising B cells and CD3+, CD4+ T cells, with few CD8+ T cells. CD21+ cells (most likely follicular dendritic cells) were also present in the dermis. Based on past and present observations, we raise an unifying hypothesis to explain the diverse extrathyroidal manifestations of Graves' disease and their apparent lack of association with TSHR autoantibodies. As opposed to the present concept that these phenomena relate to site-specific properties on preadipocytes or fibroblasts, we suggest that clinically evidence GO and dermopathy are primarily caused by local factors (particularly in the orbit) superimposed on a systemic, low-grade connective tissue inflammation. PMID- 11014314 TI - Atypical hypothyroidism and the very low birthweight infant. AB - Results of thyroid screening tests were examined retrospectively on 311,282 infants born in Massachusetts from January 1, 1993 to December 31, 1996. During this period, 118 infants were found to have typical hypothyroidism, characterized by a low thyroxine (T4) and an elevated thyrotropin (TSH) on the initial newborn screening specimen. Of these, 98 were normal birthweight (NBW, > or = 2,500 g), 9 were low birthweight (LBW, 1,501-2,499 g), and 11 were very low birthweight (VLBW, < or = 1,500 g). Atypical hypothyroidism as defined here is characterized by a low T4 and normal TSH concentration on the initial screening specimen, followed by and elevated TSH level on a repeat blood specimen. This phenomenon occurred in 18 infants, of whom 4 were NBW, 4 were LBW, and 10 were VLBW. The incidence of combined typical and atypical hypothyroidism was: NBW, 1:3051; LBW, 1:1589; VLBW, 1:153, with the highest incidence of atypical hypothyroidism in the VLBW category (48% of cases in this weight category, 56% of all cases of atypical hypothyroidism). In addition, screening programs using a primary TSH screen will miss infants with atypical hypothyroidism. In view of these results, it is suggested that T4 measurements be obtained routinely in all LBW and VLBW infants, with additional routine repeat blood specimens. PMID- 11014315 TI - Review of thyroid cancer cases among patients with previous benign thyroid disorders. AB - A previous register linkage study showed an increased risk of thyroid cancer among patients previously discharged from a hospital with a diagnosis of a benign thyroid disorder. In this study, we have reviewed all available medical records, first to validate the earlier result and second to describe the symptomatology of patients with a history of benign thyroid disorder prior to the cancer diagnosis. The previous study identified 189 patients with a benign and subsequent malignant thyroid disorder. Medical records were obtainable for 156 of these patients and were reviewed. For 104 patients, benign and malignant thyroid diseases were metachronous (a clearly separated disease history of the benign and malignant diseases), and for 48 patients synchronous. In 4 cases, thyroid cancer could not be confirmed. Among patients with metachronous thyroid disorders, all major benign thyroid disorders were represented including hot nodules, diffuse and multinodular toxic and nontoxic goiter. Symptoms preceding diagnosis of thyroid cancer included growth of goiter/nodules, globulus, stridor, hoarseness, and metastasis. No major differences were found among patients with metachronous and synchronous benign and malignant thyroid disorder, apart from the fact that all metastases were found among metachronous cases. This study confirmed the conclusion that patients with a previous history of goiter or nodules have an increased risk of thyroid cancer. However, thyroid cancer still occurs too infrequently to warrant screening in all patients with a previous history of goiter or nodules. PMID- 11014316 TI - Changes in cytokine production during pregnancy in patients with Graves' disease. AB - In order to investigate the role of type 1 and type 2 cytokines in the remission of Graves' disease (GD) during pregnancy, spontaneous and mitogen-stimulated production of interleukin (IL)-4, IL-6, IL-10, IL-12, interferon-gamma (IFN gamma), and tumour necrosis factor-alpha (TNF-alpha) were measured by enzyme linked immunospot assay of peripheral blood mononuclear cells from 10 pregnant women with GD, 8 healthy pregnant women, and 10 healthy nonpregnant women. Tests were performed in the first, second, and third trimesters of pregnancy and 10-17 weeks after delivery. IL-4 production was not affected greatly by normal or GD pregnancy, whereas IFN-gamma production was suppressed throughout pregnancy but returned to normal levels after delivery in both controls and patients. IL-6 and TNF-alpha tended to be higher in GD pregnancy than normal pregnancy, especially in the second and third trimesters. Controls had raised IL-10 in the first trimester with a return to normal levels by the third trimester, whereas patients had raised levels throughout pregnancy. IL-12 levels were suppressed to a greater extent in control than Graves' pregnancy, especially during the second and third trimesters. Ratios of IL10:IL12 in phytohemaglutinin (PHA)-stimulated cultures were much lower in GD than normal pregnancy and cross-regulation of IL-10 and IL 12 may be deficient in GD. PMID- 11014317 TI - Risk factors for malignancy of thyroid nodules initially identified as follicular neoplasia by fine-needle aspiration: results of a prospective study of one hundred twenty patients. AB - Indeterminate or suspicious findings on fine-needle aspiration (FNA) of nodular thyroid disease (i.e., findings that neither give immediate indication for surgery nor lead to clear-cut conservative management) have been the key diagnostic problem in thyroid cytology for which the inability to differentiate cytologically benign from malignant follicular growth has been one reason. The aim of this cohort study of 120 consecutive (103 females, 17 males) patients with palpable nodular thyroid disease diagnosed as follicular neoplasia (FN) by FNA (defined by the triad of high numbers of follicular cells, microfollicular arrangement, and scanty or absent colloid) was to identify patients at high risk for malignancy based on the prospective evaluation of clinical features and to characterize the histologic entities of FN. Based on a 100% surgery rate we found an 18% malignancy rate (12 papillary carcinomas, 9 follicular carcinomas). Previously suggested factors with elevated risk for malignancy such as extremes of age, male gender, and large nodule size were not associated with increased risk as were cold nodules by 99mTc-scintigraphy (relative risk: 1.2, 95% confidence interval [CI] 0.4-3.3). However, hard lesions to palpation (relative risk 2.6, 95% CI: 1.2-5.6), solitary (relative risk: 2.6, 95% CI: 1.7-4.0), and hypoechoic FNs (relative risk: 3.4, 95% CI: 2.0-5.7) by ultrasound showed elevated risks of malignancy. In summary, suspicious palpation or ultrasound results may help to define a subgroup of patients with elevated risk of malignancy when FNA indicates the diagnosis of follicular neoplasm of the thyroid. PMID- 11014318 TI - Acute interstitial nephritis and fatal Stevens-Johnson syndrome after propylthiouracil therapy. AB - We report a case of acute interstitial nephritis and fatal Stevens-Johnson syndrome in a 90-year-old woman with amiodarone-induced hyperthyroidism, who had been treated for 5 weeks with propylthiouracil (PTU). On admission, the patient exhibited acute renal failure and generalized macular purpuric eruption. Acute interstitial nephritis and Stevens-Johnson syndrome were diagnosed and PTU withdrawn. Although renal function recovered after PTU withdrawal and corticosteroid therapy, the patient's condition worsened, and she died after developing multiple organ failure. Acute interstitial nephritis seems to be rarely associated with PTU therapy. Moreover, a literature review suggested that this could represent the first published report of a Stevens-Johnson syndrome developing during PTU therapy. PMID- 11014319 TI - Primary non-Hodgkin's T-cell lymphoma of the thyroid gland complicating Hashimoto's thyroiditis: case report. AB - This case report presents an extremely rare case of primary non-Hodgkin's T-cell lymphoma of the thyroid gland complicating Hashimoto's thyroiditis and discusses the clinical history, findings, treatment, and prognosis. Although the place of surgery in the treatment of thyroid lymphoma is controversial, in this case, surgery followed by three rounds of chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisone, and radiation therapy to neck and mediastinum were a very effective treatment for the disease so that no relapse has been detected during 3-year follow-up. PMID- 11014320 TI - Subclinical hyperthyroidism. PMID- 11014321 TI - Images in thyroidology. Medullary thyroid cancer arising in an adenoma. PMID- 11014322 TI - Environmental iodine deficiency: A challenge to the evolution of terrestrial life? PMID- 11014323 TI - Pharmacokinetics of orally and intravenously administered telmisartan in healthy young and elderly volunteers and in hypertensive patients. AB - A series of studies was conducted in healthy young males and healthy elderly males or females to evaluate the pharmacokinetic profile of telmisartan. In addition, two phase-II clinical trials assessed the pharmacokinetics and the safety of telmisartan in mild-to-moderate hypertensive patients. Telmisartan was given as a single oral (1-160 mg) or intravenous (10-160 mg) dose to young males. In another multiple-dose study, telmisartan 320 mg was administered orally once daily for 7 days to healthy young male subjects. Elderly subjects received oral telmisartan (20 and 120 mg) once daily for 7 days. Telmisartan doses of 10, 20, 40, 80, 120 and 160 mg were taken once daily by mild-to-moderate hypertensive patients for 7 days. Additionally, oral telmisartan (40, 80 or 120 mg) was administered once daily for 28 days to hypertensive subjects. Following oral dosing, median time to maximum plasma telmisartan concentration was 0.5 - 2 h, with maximum plasma concentrations increasing disproportionately with dose. By contrast, plasma concentrations were directly related to the intravenous dose. Steady state was observed after 5-7 days of once-daily administration, and there was no clinically relevant accumulation at 28 days. The plasma concentration-time profiles were similar in all study groups and were characterized by fast absorption and a rapid biexponential decline after the peak plasma concentration, with a prolonged terminal elimination phase (> 20 h in healthy and hypertensive subjects). Telmisartan was well tolerated, with a low incidence of drug-related adverse events. The most frequent event was headache, which also occurred in placebo-treated control subjects. No changes in heart rate, electrocardiograms or clinical chemistry were detected following receipt of telmisartan. The study thus shows that high systemic levels of telmisartan, which are well tolerated, can be attained in healthy adults of any age and in hypertensive subjects. The long terminal elimination half-life makes telmisartan suitable for once-daily dosing and contributes to the sustained efficacy over the full 24-h dosing interval. PMID- 11014324 TI - Vacuolar hepatocyte degeneration induced by infusion of 20% glucose solution with insulin after hepatopancreatectomy in rats. AB - To investigate the causes of hepatic dysfunction after extensive resection of the liver together with pancreatectomy, rats were subjected to sham operation, to 68% hepatectomy alone, to 90% pancreatectomy alone, or to 68% hepatectomy combined with 90% pancreatectomy (hepatopancreatectomy). Solutions of 5% or 20% glucose were infused post-operatively for 48 h at a constant rate (250 ml/kg body weight/day) under fasting conditions. To improve the survival rates of pancreatectomized and hepatopancreatectomized rats given 20% glucose, it was necessary to use insulin. In hepatopancreatectomized rats, infusion of 20% glucose with insulin (1 U/5 g glucose) induced prominent hepatocyte vacuolar degeneration and mitochondrial swelling, associated with reduced hepatic protein content. The severity of histological changes was proportional to the insulin dose and the activity of hepatic glucokinase, a key glycolytic enzyme. were observed in These histological changes pancreatectomized rats albeit in a milder form, but not in sham-operated or hepatectomized rats given 20% glucose nor in any rats given 5% glucose. Our results suggest that hepatopancreatectomy followed post-operatively by a high glucose load and exogenously administered insulin enhances the development of hepatocyte swelling. PMID- 11014325 TI - Standard melatonin intake and circadian rhythms of elite athletes after a transmeridian flight. AB - Many transmeridian travellers, including top athletes, regularly take melatonin to reduce negative jet lag symptoms. Standard (rather than individually tailored) doses are often used. We examined the effects of a standard dose of melatonin on the body temperature rhythms of 12 elite biathletes (eight men and four women) after an eastward transmeridian flight to an international competition. The different effects on body temperature rhythms in men versus women underscore the need for personalized dosing schedules to avoid potential undesirable consequences. PMID- 11014326 TI - Predominant Staphylococcus aureus isolated from various skin diseases. AB - We examined predominant Staphylococcus aureus isolated from lesions of various skin diseases, and during the past 3 years, 113 methicillin-sensitive and 31 methicillin-resistant S. aureus have been isolated. The predominant species isolated from almost all of the primary bacteriological cultures was S. aureus. The skin diseases from which cultures were most frequently prepared were atopic dermatitis, followed by ulcers. S. aureus was the predominant species in two thirds or more of cases of all of the different skin diseases examined. The predominant species identified, other than S. aureus, included Streptococcus species and Pseudomonas aeruginosa. Evaluation of the predominance of S. aureus is important to the determination of the severity of skin lesions and, if needed, appropriate antimicrobial therapy. PMID- 11014327 TI - Case of ulcerative colitis associated with oesophageal ulcer. AB - A case of ulcerative colitis complicated by oesophageal ulcers is reported. A woman was admitted to our hospital because of exacerbations of ulcerative colitis both in 1992 (aged 15 years) and 1995 (aged 18 years). When she was admitted in 1995 she complained of bloody diarrhoea, sore throat and pain on swallowing. Oesophagogastro-duodenoscopy revealed oesophageal ulcers. Oesophageal pH monitoring (24-h) showed no evidence of gastro-oesophageal reflux disease. After the patient was treated she with oral prednisolone showed considerable improvement clinically and endoscopically. Initial dosage was 60 mg/day, and 1 week later, the dosage was gradually dropped since the patient responded favourably. The improvement of the oesophageal lesions coincided with the remission of ulcerative colitis. The oesophageal ulcers are, therefore, thought to be an extracolonic manifestation of ulcerative colitis. PMID- 11014329 TI - Magnetic resonance assessment of acute and chronic stroke. AB - This article describes the important role of magnetic resonance imaging (MRI) in noninvasively assessing human focal ischemic stroke. Conventional MRI, diffusion weighted and/or perfusion-weighted imaging have been used to facilitate both the qualitative and quantitative evaluation of heterogeneity of ischemic brain tissue. Further, by combining 2 or more magnetic resonance parameters, tissue signature models have been developed that may be used as surrogate markers of tissue histopathology to characterize ischemic tissue as salvageable, necrotic, or tissue in transition to necrosis. Magnetic resonance tissue-signature models and results are presented. Dynamic changes in the evolution of ischemic tissue to infarction are also discussed. Recovery from acute stroke was studied with blood oxygenation level-dependent functional MRI to investigate the neural mechanisms for recovery from aphasia after stroke. PMID- 11014328 TI - Neurologic complications of cardiac surgery. AB - The average age of patients undergoing cardiac surgery and the number of comorbidities they possess will continue to increase as surgical technology advances. Toxic/metabolic encephalopathy, hemispheric strokes, hypoxic injury, and peripheral nerve lesions all can occur as a result of cardiac surgery. Therefore, an understanding of the neurologic risk, recognizable syndromes, and preventative measures will continue to be important. Careful preoperative assessment, operative risk factor reduction, and careful postoperative assessments and management may reduce the neurologic risk for cardiac surgery. PMID- 11014330 TI - Transcranial Doppler assessment of cerebral embolic disorders. AB - Transcranial Doppler ultrasound allows noninvasive monitoring of the large intracranial vessels. Microembolic signals (MES) have been observed under a variety of circumstances, especially in symptomatic carotid stenosis, high-risk cardiac conditions, and surgical procedures (such as carotid endarterectomy and cardiopulmonary bypass surgery). Technical considerations have presented numerous challenges to the interpretation of these signals, many of which appear to represent small emboli traveling in the blood flow. The technical aspects of these MES are discussed and their significance in relation to cerebral ischemia and its prevention are examined in some detail. PMID- 11014331 TI - Cardiomyopathy in AIDS: a pathophysiological perspective. AB - This report addresses issues of pathogenesis, pathophysiology, and epidemiology of an increasingly prevalent cardiomyopathy in acquired immunodeficiency syndrome (AIDS). As patient survival increases with more effective antiretroviral therapy, cardiomyopathy in AIDS will become more apparent. The interactions of cellular and organism factors in AIDS and their relationships to the development of cardiomyopathy are reviewed herein. Amongst the factors addressed in this review are: (1) comorbid conditions found with AIDS, (2) the role of inflammatory heart disease and cytokines in the development of AIDS cardiomyopathy, (3) the pathogenetic role of vascular cells and myocardial cells in the development of cardiomyopathy, (4) the role of myocardial retroviral infection in AIDS, and (5) the impact of toxicity from antiretroviral therapy on the development of cardiomyopathy. Because it is possible that more than 1 of these factors is present in a given patient inflicted with AIDS, a multifactorial pathogenesis in AIDS cardiomyopathy must be considered. PMID- 11014333 TI - Hot off the press. PMID- 11014332 TI - Current status of safety and efficacy of calcium channel blockers in cardiovascular diseases: a critical analysis based on 100 studies. AB - Recently, serious concerns have been expressed about the long-term safety of the calcium channel blockers (CCBs) as a group. Safety and efficacy are, however, ultimately linked to each other; therefore both must be evaluated especially in the therapy of angina and hypertension, the main clinical indications for CCBs. The structural, functional, and pharmacokinetic heterogeneity of CCBs means that the efficacy and dangers of one subclass, such as the short-acting dihydropyridines (DHPs), in one situation, such as unstable angina, do not necessarily apply in other clinical situations. One hundred studies are reviewed according to their methods of data collection: case series, case control, cohort, randomized controlled trials (RCTs), and meta-analyses. Large, well-designed RCTs and the meta-analyses based on these trials remain the gold standard. Observational studies, though potentially less reliable sources of information because of selection bias, may nevertheless produce hypotheses that must then be tested in RCTs. Regarding safety, both observational studies and RCTs suggest that adverse effects of CCBs may be linked to short-acting agents, specifically short-acting nifedipine. Two good studies favor the safety of verapamil, even in short-acting form. Incomplete but increasing overall evidence favors the safety of longer-acting DHPs. Heart failure remains a class contraindication to the use of all CCBs, with some exceptions. Regarding efficacy, there are positive results of RCTs with CCBs in 2 specific clinical situations, namely, verapamil in postinfarct protection in the absence of pre-existing heart failure, and 2 outcome studies on hypertension with longer acting DHPs. These results cannot automatically be applied to other clinical situations and to other CCBs. For example, there is no evidence for the safety or efficacy of DHPs used without beta blockers in postinfarct patients. In diabetic hypertensives, 2 relatively large RCTs show that the blood pressure can be reduced by DHP-based therapy in diabetics, with a reduction in hard end points. To achieve current blood pressure goals, combination therapy is almost always necessary, and in diabetics there is strong evidence that 1 essential component should be an angiotensin converting enzyme inhibitor. The future aim with CCBs must be to obtain a large database gathered from RCTs, which will give the same certainty about efficacy and safety that already holds for use of the diuretics in hypertension, beta-blockers in postmyocardial infarction patients, and the angiotensin converting enzyme inhibitors in heart failure. PMID- 11014334 TI - Do mammals make their own morphine? PMID- 11014335 TI - Nonlantibiotic antibacterial peptides from lactic acid bacteria. PMID- 11014336 TI - The global chloromethane cycle: biosynthesis, biodegradation and metabolic role. PMID- 11014337 TI - Biomimetic synthesis of alkaloids. PMID- 11014338 TI - The iron(II) and 2-oxoacid-dependent dioxygenases and their role in metabolism. PMID- 11014339 TI - Monoterpenoids. PMID- 11014340 TI - Recommendations for the medical management of osteoarthritis of the hip and knee: 2000 update. American College of Rheumatology Subcommittee on Osteoarthritis Guidelines. PMID- 11014341 TI - The role of the chondrocyte in osteoarthritis. PMID- 11014342 TI - The course of radiologic damage during the first six years of rheumatoid arthritis. AB - OBJECTIVE: To describe the radiologic course in a large cohort of patients with early rheumatoid arthritis (RA) and to analyze individual components of damage. METHODS: Five hundred two patients with recent-onset RA (disease duration <1 year) underwent annual radiologic assessment for a maximum of 6 years in this longitudinal prospective study. The study was designed to investigate the efficacy of 3 different therapeutic strategies. For the assessment of radiologic damage, radiographs of the hands and feet were scored according to the modified Sharp/van der Heijde method (SHS; range 0-448). A mean of 2.9 (range 1-7) radiographs was read per patient. RESULTS: Stable rates of progression of the SHS, erosion score, and narrowing score were found over the course of RA: the mean rates were 8.6, 5.4, and 3.2 modified Sharp units per year, respectively. The rate of progression of newly (not previously) damaged joints declined, and the rate of progression of already damaged joints (which became more damaged) increased during followup, leading to an equal contribution to progression of the SHS at 5 years. The joints of the feet, especially the fifth metatarsophalangeal joint, generally became eroded earlier and more of them became eroded compared with the joints of the hands. CONCLUSION: Radiologic damage progresses at a constant rate. In advanced disease, monitoring the progression of previously existing damage is as important as assessing new abnormalities in previously undamaged joints. Radiographs of the feet should be included in assessments of radiologic damage that are used in clinical intervention trials and daily practice. PMID- 11014343 TI - The antiinflammatory drug sulfasalazine inhibits tumor necrosis factor alpha expression in macrophages by inducing apoptosis. AB - OBJECTIVE: Sulfasalazine (SSZ) is a commonly used drug in the treatment of inflammatory diseases such as rheumatoid arthritis and Crohn's disease. In both diseases, the proinflammatory cytokine tumor necrosis factor alpha (TNFalpha) plays a prominent role. In these studies, we investigated the mechanism by which SSZ inhibits TNFalpha expression in macrophages and macrophage-like cell lines. METHODS: Monocyte-derived macrophages and several macrophage-like cell lines were exposed to SSZ in vitro, and the effect on TNFalpha expression was monitored by reverse transcriptase-polymerase chain reaction and Western blot analysis. In addition, the effects of SSZ in vivo were examined by intraperitoneally injecting mice with SSZ, after which peritoneal cells were harvested and examined using various staining methods. RESULTS: Preincubation of macrophages with SSZ, but not with methotrexate, inhibited lipopolysaccharide (LPS)-induced TNFalpha expression. Inhibition of TNFalpha expression by SSZ coincided with the induction of apoptosis, as judged by the appearance of morphologic changes typical of apoptosis, such as nuclear condensation and fragmentation. Induction of apoptosis by SSZ was confirmed by TUNEL analysis and by the detection of cleaved U1-70K, a substrate of caspase 3. Intraperitoneal injections of SSZ in mice resulted in the induction of apoptosis of peritoneal cells within a few hours. SSZ-induced cleavage of the U1-70K protein was inhibited by Zn2+ and by specific inhibitors of caspases 3 and 8, but not caspases 1 and 9. Interestingly, the reduced expression of LPS-induced TNFalpha in the presence of SSZ was restored by inhibition of caspase 8. CONCLUSION: Inhibition of TNFalpha expression in macrophages by SSZ is due to the induction of apoptosis and involves the activation of caspase 8. PMID- 11014344 TI - Synovial macrophage depletion with clodronate-containing liposomes in rheumatoid arthritis. AB - OBJECTIVE: To assess whether intraarticular (IA) administration of clodronate liposomes results in local macrophage depletion in patients with rheumatoid arthritis (RA). Primary goals were to address both the immunohistologic and potential toxic effects of this approach. Moreover, the correlation between immunohistologic findings and clinical assessments of disease activity and cartilage damage were assessed. METHODS: An open study was conducted in consecutive RA patients who were scheduled for knee joint replacement in our department. Synovial biopsy tissue was obtained from the knee joint at 2 weeks before and at the time of surgery. This protocol was controlled for safety and immunohistologic concordance in 6 patients. One week before surgery, 10 patients received a single IA dose of clodronate liposomes. Staining of synovial tissue for cell markers (CD68, CD14, CD3, CD38) and adhesion molecules (vascular cell adhesion molecule 1 [VCAM-1], intercellular adhesion molecule 1 [ICAM-1]) was assessed by 2 blinded observers. Local and systemic parameters of disease activity were measured before each intervention. Cartilage damage was scored using standard radiologic techniques at baseline and during surgery. RESULTS: A single IA dose of clodronate liposomes significantly reduced the number of CD68 positive cells (P = 0.005) and the expression of ICAM-1 and VCAM-1 in the synovial lining (P = 0.013 and P = 0.039, respectively). The intervention did not affect fibroblast-like synoviocytes, T cells, or plasma cells. No immunohistologic changes were observed in the control group. The procedure was well tolerated. The levels of ICAM-1 and VCAM-1 in the sublining layers correlated with the extent of macroscopic synovitis (P < 0.0005 and P < 0.005, respectively). The expression of ICAM-1 and CD14 in the sublining correlated with the levels of C-reactive protein (P < 0.0005 and P < 0.01, respectively). Cartilage destruction was correlated only with the expression of CD68 in the sublining (P = 0.02). CONCLUSION: A single IA administration of clodronate liposomes leads to macrophage depletion and decreased expression of adhesion molecules in the synovial lining in patients with longstanding RA. The procedure is well tolerated, and its therapeutic potential is currently under investigation. The expression of adhesion molecules in the sublining layers reflects ongoing inflammation. PMID- 11014345 TI - Efficacy and safety of alendronate for the treatment of osteoporosis in diffuse connective tissue diseases in children: a prospective multicenter study. AB - OBJECTIVE: Osteopenia/osteoporosis is being increasingly reported as a complication of many chronic diseases, even in children. In this preliminary study, we evaluated the effect of an oral bisphosphonate (alendronate) on bone mass in children with diffuse connective tissue diseases. METHODS: Thirty-eight children with low bone mass were treated with alendronate for 1 year; 38 children who had the same primary disorders as the study patients but in a less severe form served as untreated control patients. We were also able to evaluate changes in bone mass (before and after alendronate) in 16 of the treated patients whose bone mineral density (BMD) had been routinely measured before the present study was initiated. RESULTS: BMD increased by a mean +/- SD of 14.9 +/- 9.8% (P < 0.002 versus baseline) in the treated patients (reaching the normal range in 13 patients), while the BMD was 2.6 +/- 5% (not significant versus baseline) in the control group (15 had a decrease). Most interestingly, there was a large increase in BMD (15.3 +/-9.9%) after alendronate therapy in the 16 children who had their BMD followed up in the year before the study, during which time they had shown little increase in BMD (1.03 +/- 6.3%), and often a decrease. Considering their condition, increases in the height of all patients was satisfactory. No new fractures were observed after alendronate therapy was initiated. CONCLUSION: Bisphosphonates can be considered essential components of the treatment of secondary osteoporosis, not only in adults, but also in pediatric patients. Alendronate has a positive effect on secondary osteopenia/osteoporosis in children with connective tissue diseases. PMID- 11014346 TI - Should postmenopausal women with rheumatoid arthritis who are starting corticosteroid treatment be screened for osteoporosis? A cost-effectiveness analysis. AB - OBJECTIVE: To evaluate the cost-effectiveness of different strategies for preventing corticosteroid-induced osteoporosis. METHODS: Simulated cohorts of postmenopausal women with rheumatoid arthritis (RA) starting corticosteroid treatment were examined. A Markov decision analysis model was developed to compare different management strategies, including watchful waiting, screen and treat, and empirical treatment. Treatment thresholds for the screen and treat strategy were varied from bone mineral density (BMD) T scores <-1.0 to BMD T scores <-4.0. RESULTS: Compared with a watchful waiting approach, the incremental cost-effectiveness ratio for a strategy of screen and treat with alendronate at a BMD T score of <-1.0 was $92,600 per quality-adjusted life year (QALY) gained. This result was sensitive to the cost and efficacy of osteoporosis therapy and, importantly, to the treatment threshold. At a treatment threshold of a BMD T score <-2.5, the incremental cost-effectiveness ratio of screening and treating was $76,100 per QALY. None of these results differed substantially for women taking estrogen replacement therapy. CONCLUSION: The incremental cost effectiveness ratio of a strategy of screening and treating postmenopausal female RA patients with BMD T scores of < -1.0, compared with watchful waiting, was greater than that of other well-accepted medical interventions. The cost effectiveness ratios were more acceptable when a T score treatment threshold of < 2.5 was used. These conclusions are limited by the lack of data on fracture and treatment efficacy in corticosteroid-treated patients. PMID- 11014347 TI - Interleukin-10 blockade corrects impaired in vitro cellular immune responses of systemic lupus erythematosus patients. AB - OBJECTIVE: Many systemic lupus erythematosus (SLE) patients display impaired cellular immune responses against allo- or recall antigens. Given the down regulating properties of interleukin-10 (IL-10) on antigen-presenting cell functions, this study was undertaken to investigate whether the well-known overproduction of IL-10 by SLE peripheral blood mononuclear cells (PBMC) was involved in this process. METHODS: We measured the proliferation of SLE or control PBMC against irradiated allogeneic dendritic cells in the absence or presence of antibodies blocking IL-10 activity, or in the absence or presence of IL-12. RESULTS: As a group, SLE PBMC proliferated against allogeneic targets less than control PBMC. However, SLE patients could be categorized as good responders or poor responders according to the amplitude of their allogeneic response. Interestingly, serum IL-10 concentrations were significantly higher in the poor responders than in the good responders or in the controls, and addition of antibodies blocking IL-10 activity significantly increased the proliferative responses of the group. We confirmed the role of IL-10 in the impaired allogeneic responses displayed by SLE PBMC by demonstrating that addition of IL-10 containing SLE PBMC supernatants inhibited a normal allogeneic response between unrelated healthy controls, and by showing that this inhibitory effect was commensurate with the concentrations of IL-10 measured in the supernatants. In this experimental setting, we also demonstrated that IL-10-containing SLE PBMC supernatants inhibited IL-12 p35 and IL-12 p40 gene expression. Consistent with the last observation, we found that addition of exogenous IL-12 restored the proliferation of poor-responder SLE patients' PBMC. CONCLUSION: Taken together, these results indicate that dysregulation of the IL-10/IL-12 balance plays a critical role in the impaired cellular immune responses observed in SLE patients. PMID- 11014348 TI - Association of autoantibodies against the phosphatidylserine-prothrombin complex with manifestations of the antiphospholipid syndrome and with the presence of lupus anticoagulant. AB - OBJECTIVE: To clarify the association of autoantibodies against prothrombin with the clinical manifestations of the antiphospholipid syndrome (APS) and with the presence of lupus anticoagulant (LAC). METHODS: We examined 265 patients who visited our autoimmune disease clinic. IgG and IgM antiprothrombin antibodies were tested by enzyme-linked immunosorbent assay (ELISA) as either antiphosphatidylserine-prothrombin complex (aPS/PT) antibodies or as antibodies against prothrombin coated on irradiated ELISA plates (as antigen) (aPT). IgG, IgM, and IgA anticardiolipin (aCL) antibodies and their beta2-glycoprotein I (beta2GPI) dependency were also evaluated by ELISA. LAC was tested by 3 different methods. RESULTS: The presence of aPS/PT, but not of aPT, significantly correlated with the clinical manifestations of APS (odds ratio [OR] 4.39, 95% confidence interval [95% CI] 2.06-9.38), and aPS/PT antibodies were as specific as beta2GPI-dependent aCL for APS (93.1% for both). IgG aPS/PT strongly correlated with the presence of LAC as detected using the dilute Russell viper venom time test (OR 38.2, 95% CI 13.4-109.1). CONCLUSION: Antiprothrombin antibodies are heterogeneous and their clinical relevance depends on the method of detection applied. Positive results on the aPS/PT test can serve as a marker of thrombotic events in patients with autoimmune diseases. PMID- 11014349 TI - Chlorpromazine induces apoptosis in activated human lymphoblasts: a mechanism supporting the induction of drug-induced lupus erythematosus? AB - OBJECTIVE: Drug-induced lupus erythematosus is a serious side effect of certain medications, such as procainamide, quinidine, hydralazine, chlorpromazine, and isoniazid, the underlying pathogenesis of which is unresolved. In this study, we examined the influence of these drugs on the regulation of apoptosis, or programmed cell death, in quiescent and activated human lymphocytes. We also discuss the dysregulation of apoptosis as a pathogenetic factor in systemic lupus erythematosus. METHODS: Peripheral blood mononuclear cells or activated lymphoblasts from normal donors were incubated with different concentrations of each of the above-mentioned drugs. RESULTS: We did not find induction of apoptosis in quiescent cells over a broad concentration range. In contrast, lymphoblasts readily underwent apoptosis when cultured with chlorpromazine, but not any of the other drugs, after stimulation with interleukin-2 (IL-2) in a dose , time- and cell cycle-dependent manner. By several lines of evidence, toxicity was ruled out. Characteristic features of apoptosis-like incorporation of propidium iodide (PI), such as increased annexin V binding, changes in mitochondrial membrane potential, and induction of DNA breaks (as evidenced by TUNEL techniques), could be induced in lymphoblasts after chlorpromazine treatment. Chlorpromazine did not cause apoptosis by inhibition of cytokine binding or blockade of early intracellular signaling. The protease inhibitor Z VAD and the ceramide inhibitor sphingosine 1-phosphate effectively blocked chlorpromazine-induced apoptosis (by PI staining and by externalization of phosphatidylserine), in contrast to the caspase 3/CPP32 inhibitor DEVD, which had only minor effects. Western blot analysis revealed IL-2-mediated phosphorylation of extracellular signal-regulated kinase, which was sensitive to chlorpromazine. Using lymphoblasts from a patient with Canale-Smith syndrome, we found that chlorpromazine-mediated apoptosis is Fas/ APO-1 independent. CONCLUSION: These data suggest that chlorpromazine mediates apoptosis in human lymphoblasts through specific activation of intracellular proapoptotic signaling cascades. This mechanism might lead to an unsynchronized inflow of apoptotic break-down products and thereby to the induction of (auto)immunity against nuclear components. PMID- 11014350 TI - HLA-DQA1*0501 is associated with diffuse systemic sclerosis in Caucasian men. AB - OBJECTIVE: Systemic sclerosis (SSc) is uncommon in men, and relatively little is known about factors contributing to its pathogenesis in this population. In the current study, we investigated HLA class II alleles in men with SSc. We also investigated the hypothesis that HLA compatibility of the mother could be a risk factor for SSc in men. METHODS: Sequence-specific oligonucleotide probe typing was used to determine DQA1, DQB1, and DRB1 alleles of SSc patients (50 men and 36 parous women), healthy controls (59 men and 80 parous women), 26 mothers of men with SSc, and 44 mothers of healthy men. All study subjects were Caucasian, and allele frequencies were compared with those of Caucasian controls from the Eleventh International Histocompatibility Workshop as well as those of local controls. RESULTS: The DQA1*0501 allele was significantly increased among men with SSc compared with healthy men (odds ratio [OR] 2.3, P = 0.006, Pcorr = 0.04). DQA1*0501 was associated with diffuse SSc in men (OR 3.0, P = 0.004, Pcorr = 0.03), but not with limited SSc in men. Maternal HLA compatibility was not a risk factor for SSc in men. CONCLUSION: Previous studies have shown associations of DRB1 alleles with SSc, but have rarely determined DQA1 allele frequencies. Our findings indicate that a specific DQA1 allele is associated with SSc, and that DRB1 associations may be due to linkage disequilibrium with DQA1. Moreover, by analyzing genetic susceptibility according to sex, we found that the contribution of HLA genes to the risk of SSc was substantially greater in men than in parous women. PMID- 11014351 TI - Histopathologic evidence that sacroiliitis in ankylosing spondylitis is not merely enthesitis. AB - OBJECTIVE: To systematically study the histopathology of sacroiliitis in ankylosing spondylitis (AS) at 5 different stages of the disease. METHODS: Two independent observers assessed 75 microscopic features in the sacroiliac (SI) joints in 12 cases of AS (5 biopsies, 7 autopsies) and in 22 control cases (all autopsies). RESULTS: In AS, synovitis, pannus formation, myxoid marrow, superficial cartilage destruction, enthesitis, intraarticular fibrous strands, new bone formation, and bony ankylosis were significantly more frequent than in control cases, in which there was more endochondral bone within deep-zone articular cartilage. Cartilaginous fusion occurred in both groups, but much earlier in AS. There was no residual synovium when the joint lumen was totally occluded. Mild but destructive synovitis and myxoid subchondral bone marrow were the earliest changes identified in AS. These lesions destroyed the adjacent articular tissues, a loss that was followed to varying degrees by fibrous scarring, woven bone, and new cartilage. The original cartilages also fused, and chondral fusion was the predominant mode of ankylosis. Both the original and the reparative cartilaginous tissues were replaced by bone. Active enthesitis occurred in 2 advanced and 3 late cases; fibrous scar tissue, presumed to represent previous enthesitis, was observed in all stages except the earliest. Paraarticular bone was at first dense, and later porotic. CONCLUSION: In the sacroiliitis of AS, two findings predominate: 1) synovitis and subchondral bone marrow changes offer a more rational explanation for widespread joint destruction than does enthesitis; and 2) an unusual form of chondroid metaplasia contributes to ankylosis. PMID- 11014352 TI - Prediction of relapses in Wegener's granulomatosis by measurement of antineutrophil cytoplasmic antibody levels: a prospective study. AB - OBJECTIVE: Prediction of relapses in Wegener's granulomatosis (WG) by measuring levels of antineutrophil cytoplasmic antibodies (ANCA) directed against proteinase 3 (PR3) or myeloperoxidase (MPO) remains a controversial issue. To assess the value of serial quantification of ANCA by indirect immunofluorescence (IIF) and antigen-specific enzyme-linked immunosorbent assay (ELISA) for monitoring disease activity in patients with WG, a prospective observational study was conducted in patients with WG attending an outpatient clinic in the Netherlands. METHODS: One hundred patients with WG (85 with PR3-ANCA, 15 with MPO ANCA) were studied prospectively from 1996 to 1998. Serum samples were obtained and analyzed every 2 months for ANCA levels. Disease activity was prospectively assessed without knowledge of the ANCA levels. RESULTS: Relapses occurred in 37 of 100 patients (37%). Thirty-four (92%) of the 37 patients showed a rise in the level of ANCA preceding their relapse, as detected by ELISA or IIF. The predictive value of an increase in ANCA titers for relapse was 57% (17 of 30) for cytoplasmic/classic ANCA (cANCA; by IIF), 71% (27 of 38) for PR3-ANCA (by ELISA), and 100% (3 of 3) for MPO-ANCA (by ELISA). The predictive value of a rise in ANCA as measured by ELISA or IIF did not substantially improve following concomitant measurement of the IgG3 subclass of PR3-ANCA. Forty-three percent of patients who showed a rise in cANCA (by IIF) and 29% with a rise in PR3-ANCA (by ELISA) did not subsequently experience a relapse. CONCLUSION: Serial measurement of ANCA levels is valuable for the early prediction of relapses in patients with WG. PMID- 11014353 TI - An autosomal dominant periodic fever associated with AA amyloidosis in a north Indian family maps to distal chromosome 1q. AB - OBJECTIVE: To investigate genetic susceptibility in the first Indian family identified as having an autosomal dominantly inherited periodic fever syndrome. The inflammatory disease was characterized chiefly by arthralgia, skin rashes, and AA amyloidosis. METHODS: Markers from known periodic fever susceptibility loci were investigated in 7 affected and 11 healthy members of a north Indian family. These included the TNFRSF1A locus (formerly known as TNFRI), which is involved in autosomal dominant tumor necrosis factor receptor-associated periodic syndrome on chromosome 12p13, the familial Mediterranean fever locus (MEFV) on chromosome 16p13, the hyperimmunoglobulinemia D and periodic fever syndrome (HIDS) locus on chromosome 12q24, and the Muckle-Wells syndrome/familial cold urticaria (MWS/FCU) locus on distal chromosome 1q44. RESULTS: Linkage to both TNFRSF1A and MEFV was definitively excluded, and DNA sequencing of these genes revealed no mutations. Furthermore, there was no evidence of linkage to the HIDS locus. In contrast, significant logarithm of odds scores for 5 markers from the MWS/FCU region were obtained in this family, and the disease segregated with the same haplotype in all affected members. CONCLUSION: We have identified an inherited inflammatory disease in a north Indian family with clinical features overlapping some of those of MWS and FCU. The susceptibility gene maps to distal chromosome 1q44, a region already implicated in both MWS and FCU. Different mutations in the same (or a closely related) gene may be responsible for an inflammatory disease with a broad phenotype among diverse ethnic populations. PMID- 11014354 TI - Localization of a gene for familial recurrent arthritis. AB - OBJECTIVE: To localize the gene for familial recurrent arthritis via a genome wide linkage scan in an extended kindred with the disease. METHODS: A 3 generation family in which 9 members were diagnosed with juvenile idiopathic arthritis (JIA) was ascertained. In this family the disease was of very early onset and included episodic inflammation leading to eventual destruction of joints, muscle, and skin. We treated this disorder as a distinct clinical entity that we have named "familial recurrent arthritis." A genome-wide linkage scan with polymorphic microsatellites at 10-15-cM resolution was initiated. RESULTS: The genome-wide scan generated a maximum 2-point logarithm of odds score with D15S211 (Zmax = 3.27 at thetamax = 0.0010). Haplotype reconstruction defined a candidate region of approximately 20 cM flanked proximally by D15S983 and distally by D15S127 on human chromosome 15. CONCLUSION: A gene for familial recurrent arthritis was localized to 15q22-24, as a result of a genome-wide linkage scan in a large, multiply affected kindred. Identification of the altered gene will provide insights into the pathogenesis of autoimmune joint destruction that is reminiscent of JIA. PMID- 11014355 TI - Clinical images: Neuropsychiatric systemic lupus erythematosus. PMID- 11014356 TI - Mesenchymal cells expressing bone morphogenetic protein receptors are present in the rheumatoid arthritis joint. AB - OBJECTIVE: To evaluate the presence of cells of an early mesenchymal lineage, as judged by the expression of bone morphogenetic protein receptors (BMPRs), in the joints of normal individuals and patients with rheumatoid arthritis (RA). METHODS: Synovial fluids, single cell suspensions of cultured fibroblast-like synoviocytes (FLS), and synovial tissues were examined by immunohistology with antibodies to BMPR type IA (BMPRIA), BMPRIB, and BMPRII and then quantified using computerized image analysis. Other antibodies were evaluated by cytofluorography. RESULTS: In primary cultures of joint effusions from patients with RA and other forms of inflammatory arthritis, there were large adherent cells with the appearance of either fibroblasts or stromal cells that stained with antibodies to mesenchymal elements-CD44, type I collagen, alpha-actin, and vimentin-but not with antibodies to hematopoietic markers. These cells proliferated rapidly, expressed BMPRIA and BMPRII, and soon became the predominant cells in culture. They were retained through multiple passages and persistently displayed surface vascular cell adhesion molecule 1. Immunohistochemical analysis of cultured RA FLS (passages 3, 4, and 6; n = 6) revealed that 11.6% were BMPR-positive, while only 2.0% of osteoarthritis FLS (passage 4; n = 3) were BMPR-positive, and 1 normal synovial culture had no BMPR-positive cells. In all RA synovial membranes examined (n = 9), BMPRI- and BMPRII-expressing cells were identified in the intimal lining and were also scattered in the subintima. These cells constituted approximately 25% and approximately 7% of the cells in each area, respectively. Double immunostaining showed no coexpression of BMPR-positive cells with CD68, CD34, or CD3. Cells expressing BMPR were not seen in any normal synovial samples (n = 4). Strong staining for BMPR was identified on cells at the invasive front of the pannus and at sites of cartilage erosion. CONCLUSION: The inflamed RA joint contains BMPR-positive mesenchymal cells. Their origin is still speculative, but since their counterparts in the bone marrow are essential for osteoclastogenesis, support lymphocyte development and maturation, and protect T cells and B cells from programmed cell death, the BMPR-positive cells may be essential elements in the pathogenesis of RA and other inflammatory forms of chronic synovitis. PMID- 11014357 TI - Suppression of arthritis and protection from bone destruction by treatment with TNP-470/AGM-1470 in a transgenic mouse model of rheumatoid arthritis. AB - OBJECTIVE: We assessed the clinical and histologic features of angiogenesis inhibition in a transgenic mouse model of arthritis that closely resembles rheumatoid arthritis (RA) in humans. METHODS: KRN/NOD mice, which spontaneously develop arthritis, were treated with TNP-470, an angiogenesis inhibitor. Disease was monitored by use of clinical indices and histologic examinations; circulating blood levels of vascular endothelial growth factor were determined by enzyme linked immunosorbent assay. RESULTS: In the preventive protocol, with TNP-470 administration at a dosage of 60 mg/kg of body weight, the onset of arthritis was delayed and its clinical intensity was rather mild; 100% of placebo-treated transgenic mice developed arthritis that led to severe articular destruction. At a dosage of 90 mg/kg of TNP-470, the appearance of clinical signs was delayed for a longer period of time and disease was almost abolished. The therapeutic regimen alleviated clinical signs only when given during the very early stage of disease. Reductions in cartilage and bone destruction by TNP-470 treatment were observed histologically, a feature that was still evident at 30 and 80 days after injections were withdrawn. CONCLUSION: Our demonstration that in vivo administration of an angiogenesis inhibitor suppresses arthritis and protects from bone destruction provides new insight into the pathogenesis of the disease and opens new possibilities in the treatment of RA in humans. PMID- 11014358 TI - Suppression of streptococcal cell wall-induced arthritis by human chorionic gonadotropin. AB - OBJECTIVE: To determine whether human chorionic gonadotropin (HCG) contributes to pregnancy-associated immunosuppression, as observed clinically by an amelioration of symptoms in human autoimmune diseases, including rheumatoid arthritis, during pregnancy. METHODS: Administration of HCG was initiated 2 days prior to an arthritogenic dose of streptococcal cell wall (SCW) in nonpregnant female rats, and the development and severity of SCW-induced arthritis was monitored. Inflammatory mediators, including plasma nitrite/nitrate and cytokine levels, were measured. Inducible nitric oxide synthase (iNOS) protein and cytokine messenger RNA expression in joint tissue were compared between treated and untreated arthritic animals. RESULTS: Systemic administration of HCG resulted in a dose-dependent reduction in the clinical arthritis index. Consistent with the amelioration of clinical symptoms, HCG significantly reduced the inflammatory cell infiltration, pannus formation, and bone and cartilage degradation. Mechanistically, HCG therapy was associated with suppression of the overzealous production of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-1beta, which contribute to synovial pathology in animals with SCW-induced arthritis. Circulating nitric oxide and the amount of iNOS protein were also reduced. Furthermore, circulating transforming growth factor beta levels were elevated by the HCG, all of which suggest monocytes/macrophages as a potential target. CONCLUSION: These findings indicate that HCG exerts a protective effect in this experimental arthritis model, through modulation of inflammatory mediators. PMID- 11014359 TI - The effects of local administration of lactoferrin on inflammation in murine autoimmune and infectious arthritis. AB - OBJECTIVE: To determine whether lactoferrin can modify articular inflammation in murine models of autoimmune and septic arthritis. METHODS: Collagen arthritis was induced in DBA/1 mice and Staphylococcus aureus septic arthritis in Swiss mice. Joints with established inflammation were injected periarticularly with 0.5 mg or 1 mg of human lactoferrin, and arthritis was monitored for 3 days. RESULTS: DBA/1 mice injected with lactoferrin showed significantly suppressed local inflammation for up to 3 days, achieving up to 71% of the effect of corticosteroid. Periarticular injection of 125I-lactoferrin confirmed that 25% of lactoferrin was retained in paws after 6 hours. Serum levels of interleukin-6, however, were not significantly reduced, suggesting a predominantly local antiinflammatory effect. Similarly, local, periarticular administration of lactoferrin into S aureus infected Swiss mice significantly suppressed paw inflammation and did not enhance bacterial survival. CONCLUSION: Lactoferrin may have clinical utility in reducing articular inflammation, particularly in septic arthritis, in which antiinflammatory effects may be achieved without promoting bacterial survival. PMID- 11014360 TI - Functional estrogen receptors in adult articular cartilage: estrogen replacement therapy increases chondrocyte synthesis of proteoglycans and insulin-like growth factor binding protein 2. AB - OBJECTIVE: Epidemiologic studies suggest a protective effect of estrogen replacement therapy (ERT) against the development of knee and hip osteoarthritis, but a potential mechanism for this effect is not known. The present study was done to determine if functional estrogen receptors (ERs) are present in adult articular cartilage and to determine if ERT in vivo affects the production of insulin-like growth factor binding proteins (IGFBPs). METHODS: Reverse transcription-polymerase chain reaction, immunoblotting, and immunohistochemistry were used to measure messenger RNA (mRNA) and protein for ERs in adult monkey articular cartilage. Cultured chondrocytes transfected with a reporter construct containing the estrogen response element (ERE/luciferase) were stimulated with estrogen in vitro to determine functional activity of the ERs. IGFBP production was measured by ligand and immunoblotting of conditioned media of cells cultured from control and estrogen-treated surgically menopausal monkeys. Proteoglycan (PG) synthesis was estimated by measurement of 35SO4 incorporation. RESULTS: ERa and ERbeta mRNA were present in adult monkey articular cartilage, and ER protein was demonstrated by immunoblotting and immunohistochemistry. Estrogen treatment in vitro of cells transfected with the ERE/luciferase construct resulted in a 2.87-fold increase (P = 0.0163) in reporter production over that of untreated cells. Compared with untreated controls, IGFBP-2 production was significantly increased (P < 0.008) in conditioned media of chondrocytes cultured from monkeys that had received ERT in vivo. Increased IGFBP-2 in these cultures was associated with a 1.41-fold increase (P = 0.02) in the level of sulfate incorporation. CONCLUSION: Transcriptionally functional ER are present in adult articular cartilage, and ERT increases the production of IGFBP-2 and the synthesis of PGs by chondrocytes from surgically menopausal monkeys. These results indicate that estrogen can have a direct effect on adult articular cartilage. PMID- 11014361 TI - Mechanotransduction via integrins and interleukin-4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal, but not osteoarthritic, human articular chondrocytes. AB - OBJECTIVE: To determine molecular events in the regulation of messenger RNA (mRNA) of cartilage matrix molecules and proteases by mechanical stimulation of chondrocytes from normal human articular cartilage and to ascertain whether similar regulatory systems are present in chondrocytes from osteoarthritic (OA) cartilage. METHODS: Chondrocytes extracted from macroscopically and microscopically normal and OA cartilage were mechanically stimulated in the presence or absence of GRGDSP or GRADSP oligopeptides, neutralizing interleukin-4 (IL-4) antibodies, gadolinium, or apamin. The relative levels of mRNA for aggrecan, tenascin, matrix metalloproteinase 1 (MMP-1), MMP-3, and tissue inhibitor of metalloproteinases 1 (TIMP-1) were determined by semiquantitative reverse transcription-polymerase chain reaction at several time points up to 24 hours poststimulation, using GAPDH as a control. RESULTS: Normal chondrocytes showed an increase in aggrecan mRNA and a decrease in MMP-3 mRNA within 1 hour following stimulation, with a return to baseline levels within 24 hours. These changes were blocked by GRGDSP, IL-4 antibodies, and gadolinium, but were unaffected by apamin. In contrast, chondrocytes isolated from OA cartilage showed no change in aggrecan or MMP-3 mRNA levels following mechanical stimulation. The mRNA levels of tenascin, MMP-1, and TIMP-1 were unaltered in mechanically stimulated normal and OA chondrocytes. CONCLUSION: Mechanical stimulation of human articular chondrocytes in vitro results in increased levels of aggrecan mRNA and decreased levels of MMP-3 mRNA. The transduction process involves integrins, stretch-activated ion channels, and IL-4. This chondroprotective response is absent in chondrocytes from OA cartilage. Abnormalities of mechanotransduction leading to aberrant chondrocyte activity in diseased articular cartilage may be important in the progression of OA. PMID- 11014362 TI - Modulation of collagenase 3 in human osteoarthritic cartilage by activation of extracellular transforming growth factor beta: role of furin convertase. AB - OBJECTIVE: Treatment of normal cartilage with transforming growth factor beta (TGFbeta) can increase the synthesis of collagenase 3 by chondrocytes and mimic the in situ distribution of this enzyme in osteoarthritic (OA) cartilage, which occurs predominantly in the deep zone. In this study, we examined the elements of the TGFbeta system that are potentially relevant to this effect. METHODS: TGFbeta1 and TGFbeta2 levels in cultured cartilage explants were determined by enzyme-linked immunosorbent assay (ELISA). OA cartilage explants were treated with small latent TGFbeta1 complex in the presence of various inhibitors, and collagenase 3 levels were determined by ELISA. The inhibitors were against serine proteases, plasmin, cathepsins, furin, and a neutralizing antibody against the mannose-6 phosphate/ insulin-like growth factor 2 receptor (M6P/IGF-2R). Small latent TGFbeta1, TGFbeta receptor types I, II, and III (TGFbetaRI, RII, and RIII), M6P/IGF-2R, and furin were immunolocalized in cartilage. RESULTS: Our data showed that latent TGFbeta1 is the major isoform that is synthesized; levels of 17.2 +/-1.7 pg/mg and 1.1 +/- 0.3 pg/mg tissue wet weight (mean +/- SEM) were found for total TGFbeta1 and TGFbeta2, respectively, in OA cartilage. A general serine protease inhibitor abrogated activation of both endogenous and exogenous small latent TGFbeta1. Plasmin and furin inhibitors and anti-M6P/IGF-2R reduced the levels of exogenous small latent TGFbeta1 complex-induced collagenase 3 by 33%, 95%, and 76%, respectively, but the cathepsin inhibitor had no effect. Immunolocalization of the small latent TGFbeta1 complex as well as of TGFbetaRI and RII revealed a statistically significant increase in the chondrocyte score in only the deep zone of OA cartilage. The M6P/IGF-2R level was significantly higher in OA cartilage in both the superficial and deep zones. Furin was found in normal cartilage exclusively in the superficial zone, whereas in OA cartilage, a level similar to that in normal cartilage was found in the superficial zone, but a significantly higher cell score (mean +/- SEM 23.6 +/- 4.7%) was registered in the deep zone. CONCLUSION: The mechanisms of TGFbeta activation/ activity with regard to collagenase 3 modulation in cartilage appear to be controlled by furin convertase with or without M6P/IGF-2R. These factors and the small latent TGFbeta complex are increased in the deep zone of OA cartilage, corresponding to the preferential site of collagenase 3 production. PMID- 11014363 TI - Reduction in the chondrocyte response to insulin-like growth factor 1 in aging and osteoarthritis: studies in a non-human primate model of naturally occurring disease. AB - OBJECTIVE: Although the development of osteoarthritis (OA) is closely associated with aging, the mechanism for this association is not clear. This study was designed to determine the effects of aging and OA on the chondrocyte response to stimulation with insulin-like growth factor 1 (IGF-1) in a non-human primate model of naturally occurring OA. METHODS: Chondrocytes were isolated from cartilage removed separately from the medial and lateral femoral condyles and tibial plateaus of cynomolgus monkeys at the time of necropsy. Each joint site was scored histologically on a scale of 0-7 for OA pathologic changes. Isolated chondrocytes were cultured in alginate in serum-free medium and stimulated with IGF-1 or des(1-3) IGF-1, which has a much lower affinity for IGF binding proteins (IGFBP) than IGF-1. Response was measured as the ability to stimulate sulfate and proline incorporation. RESULTS: Cartilage samples from 34 monkeys ranging in age from 6.7 years to 27 years and with histologic scores ranging from 0 to 7 were analyzed. A significant decline in the response to IGF-1 was noted with both increasing age and increasing OA score. Controlling for the OA score, the estimated effect of age on IGF-1 response, measured by total sulfate incorporation, was a decline of 3.81% per year (P = 0.0001), or a 75% decline over 20 years as a monkey ages from young to older adult. Controlling for age, the effect of OA score was significant only for proline incorporation in the alginate matrix (estimated slope coefficient +/-standard error -15.9 +/- 7.2; P = 0.03), suggesting a negative effect of OA on retention of 3H-proline-labeled proteins in the matrix. There was a significantly reduced response to des(1-3) IGF-1 with increasing age, but no effect of OA score on response to des(1-3) IGF 1. There was no effect of age on cell viability. CONCLUSION: These results demonstrate a significant age-related decline in the chondrocyte response to IGF 1. The finding that increasing OA score was associated with a reduced response to intact IGF-1 but not des(1-3) IGF-1 suggests a role of increased production of inhibitory IGFBP in OA. Since the cells from older animals had a reduced response to both forms of IGF-1, the mechanism of the reduced response with age cannot be attributed to changes in IGFBP. Age-related changes in IGF receptors or, more likely, age-related alterations in intracellular signal transduction may also be involved. PMID- 11014364 TI - Differences in type II collagen degradation between peripheral and central cartilage of rat stifle joints after cranial cruciate ligament transection. AB - OBJECTIVE: Type II collagen degradation is thought to be the key process in cartilage degradation during the development of osteoarthritis (OA). In this study, we investigated the kinetics of type II collagen degradation during surgically induced OA. METHODS: Experimental OA was induced in male Wistar rats by transecting the cranial (anterior) cruciate ligament (CCL). Hematoxylin and eosin staining was used to study overall cartilage degradation, while immunostained sections were used to demonstrate denatured type II collagen (Col2 3/4m antibody) and the collagenase cleavage site in type II collagen (Col2-3/ 4Cshort antibody). RESULTS: During the first 3-4 weeks, cartilage destruction, associated with chondrocyte death, proteoglycan depletion, and a marked increase in the collagenase cleavage neoepitope, was mainly located at the margins of the cartilage. From weeks 3-4, the central part of the cartilage showed increased surface fibrillation and apparent chondrocyte death. In these areas, increased denatured type II collagen staining but little cleavage-site staining was present. CONCLUSION: These results indicate that cartilage degradation after CCL transection in the rat consists of 2 phases. An early phase located at the cartilage margins and a late phase located at the central part of the cartilage. In the early phase, collagenase-dependent cartilage damage occurred. During the late phase, the level of type II collagen denaturation increased. PMID- 11014365 TI - Analysis of autoimmune bone marrow by antibody-phage display: somatic mutations and third complementarity-determining region arginines in anti-DNA gamma and kappa V genes. AB - OBJECTIVE: To examine anti-double-stranded DNA (anti-dsDNA) IgG autoantibodies from the bone marrow of individuals with systemic lupus erythematosus (SLE). METHODS: A library of single-chain variable fragments (scFv) was constructed from SLE bone marrow complementary DNA of gamma, kappa, and lambda isotype by cloning into the pHENIX phagemid vector. The library was screened with dsDNA in solution, and 2 anti-DNA phage, DNA1 and DNA4, were isolated and their Ig V genes sequenced. Soluble scFv corresponding to DNA1 and DNA4, and their heavy (H)- and light (L)-chain recombinants, were prepared, purified, and analyzed for binding to DNA by enzyme-linked immunosorbent assay. RESULTS: DNA1 and DNA4 used different Ig H-chain (3-30 and 5-51, respectively) and L-chain (DPK15 and DPK22, respectively) V genes. The ratios of replacement mutations to silent mutations in DNA1 and DNA4 suggest that their V genes were selected for improved antigen binding in vivo. The recombinant between DNA4VH and DNA1VL showed the highest relative affinity for both single-stranded DNA and dsDNA. These 2 Ig subunits contained third complementarity-determining region arginines and had acquired the majority of replacement mutations. CONCLUSION: Anti-dsDNA IgG autoantibodies from the bone marrow of SLE patients exploit diverse V genes and cationic V-D-J and V J junctions for DNA binding, and accumulate replacement mutations that enhance binding. PMID- 11014366 TI - Sjogren's syndrome occurring after hepatitis B vaccination. PMID- 11014367 TI - Apolipoprotein(a) deposition in atherosclerotic coronary arteries of a patient with systemic lupus erythematosus: comment on the article by Romero et al. PMID- 11014368 TI - Presentation of the Southern Society for Clinical Investigation Founders Medal to Dr. Martin Steinberg. PMID- 11014369 TI - Southern Society for Clinical Investigation Founders Medal recipient's address. PMID- 11014370 TI - Regional hemodynamics after chronic nitric oxide inhibition in spontaneously hypertensive rats. AB - BACKGROUND: Inhibition of nitric oxide (NO) synthase by L-arginine analogs is associated with elevation of blood pressure in rats. Because endothelium dependent vasomotion in different vascular beds is not homogenous, the aim of this study was to characterize and compare regional hemodynamic responses in carotid, femoral, and renal vascular beds after chronic NO inhibition in spontaneously hypertensive rats. The possible role of circulating endothelin and renin angiotensin systems in mediating the effects of chronic NO inhibition was also studied. METHODS: Systemic and regional hemodynamics, left ventricular mass, plasma renin activity, and plasma endothelin-1 were determined in control and Nomega-nitro-Larginine methyl ester (L-NAME)-treated (10 mg/kg/day, 4 weeks) spontaneously hypertensive rats. RESULTS: L-NAME treatment increased arterial pressure and total peripheral and regional vascular resistance and decreased cardiac output, stroke volume, and regional blood flow. An increase in blood flow ratio and a decrease in vascular resistance ratio between carotid and renal as well as femoral and renal vascular beds in rats treated with L-NAME was found. Blood flow and vascular resistance ratios between femoral and carotid vascular beds remained unchanged. L-NAME increased plasma renin activity and left ventricular weight/body weight ratio, whereas plasma endothelin-1 was not modified. CONCLUSIONS: The results of this study showed that the renal circulation seemed to be more sensitive to the effects of chronic NO inhibition than carotid and femoral vascular beds. Simultaneous activation of the renin angiotensin system may further potentiate cardiovascular effects of chronic NO inhibition. No evidence that circulating endothelin-1 plays a role in this model of hypertension was found. PMID- 11014371 TI - Increased leukocyte aggregation induced by gamma-globulin: a clue to the presence of pseudoleukopenia. AB - BACKGROUND: Previous articles have reported the appearance of leukopenia after the administration of high-dose intravenous gamma-globulins. METHODS: A simple slide test was used to reveal the state of leukocyte aggregation (LA) in the peripheral blood as well as the absolute number of leukocytes in the slides. RESULTS: A significant (P < 0.00001) increment was noted in the state of LA, from 9 +/- 6% before to 25 +/- 11% after gamma-globulin administration. This was accompanied by a significant (P < 0.0004) reduction in the number of white blood cells in the peripheral blood (from 8433 +/- 3905 to 6550 +/- 3252 cells/mm3) but no significant change in the absolute number of the cells as determined by the leukocyte count per high power field in the peripheral slides. CONCLUSIONS: We raise the possibility that the leukopenia reported in some patients after the intravenous administration of high dose gamma-globulin is explained in part by the agglutination effect of the drug. PMID- 11014372 TI - Serum level of soluble tumor necrosis factor receptor II (sTNF-R75) is apparently an index of overall monocyte-related infectious and inflammatory activity. AB - BACKGROUND: The serum level of soluble tumor necrosis factor receptor II (sTNF R75) has been recently found to correlate with the activity and/or severity of several different infectious and inflammatory diseases. These results have led us to presume that the serum sTNF-R75 level reflects the active immune activity of all causes and may correlate well with nonspecific infectious and inflammatory markers such as peripheral leukocyte counts and serum C-reactive protein level. METHODS: In total, 110 apparently healthy adults, 55 men and 55 women, were enrolled in the study. Serum levels of sTNF-R75, C-reactive protein, globulin, alkaline phosphatase, lactate dehydrogenase, creatinine, urea nitrogen, and counts of neutrophils, lymphocytes, monocytes, eosinophils, and basophils were checked. The relationships between the serum sTNF-R75 level and other parameters were analyzed using the SAS statistical program. RESULTS: By various statistical methods, the serum sTNF-R75 level showed consistently significant positive links with peripheral monocyte count, serum C-reactive protein level, and two parameters of renal clearance function (serum urea nitrogen and creatinine levels). Serum levels of alkaline phosphatase and lactate dehydrogenase had significant positive links with the serum sTNF-R75 level by multivariate regression analysis. There was no significant link between the serum sTNF-R75 level and counts of neutrophils, lymphocytes, eosinophils, or basophils. CONCLUSIONS: Our results, together with those of recent reports showing positive correlations between the serum sTNF-R75 level and activities/severities of different infectious and inflammatory diseases, and also that TNF-alpha is principally produced by monocytes and macrophages, suggest that the serum sTNF R75 level is very probably an index of overall monocyte-related infectious and inflammatory activities. PMID- 11014373 TI - Proteinuria: potential causes and approach to evaluation. AB - Proteinuria may be associated with a renal or systemic disease, or it may be isolated. The latter occurs in asymptomatic patients without evidence of any disease or abnormality of the urine sediment. Isolated proteinuria may be subdivided into two broad groups: (1) benign forms, with a favorable-to-excellent prognosis and (2) persistent forms, some of which have a worrisome prognosis. Functional proteinuria may occur in disorders with altered renal hemodynamics, usually resolves, and is not associated with progressive renal disease. Idiopathic transient proteinuria is typically discovered on routine screening and usually disappears on subsequent testing. In idiopathic intermittent proteinuria, a significant number (50%) of urine samples exhibit abnormal rates of protein excretion. Although structural abnormalities may be observed on renal biopsy, progressive renal insufficiency is unusual. In orthostatic proteinuria, the rate of protein excretion completely normalizes in the recumbent position. Long-term studies show this to be a benign condition. In persistent isolated proteinuria, at least 80% of random urine samples exhibit abnormal protein excretion. This represents a heterogeneous group, but a significant proportion of these patients have prominent renal pathologic findings and progress to serious renal disease. Proteinuria with significant renal disease may be non-nephrotic or nephrotic range. The former does not exclude glomerular disease, but tubulointerstitial or vascular disorders are also likely when proteinuria is less than 2 g/24 hours. Patients with nephrotic-range proteinuria generally have a glomerular disorder. Distinction between benign and more ominous forms of proteinuria requires careful evaluation. PMID- 11014374 TI - A 65-year-old man with chronic back pain and shortness of breath. PMID- 11014375 TI - Vasodilator therapy for chronic aortic and mitral regurgitation. AB - The use of vasodilator therapy in chronic AR and MR may be beneficial in selected patients and harmful in others. The hemodynamics of the two conditions are different and must be taken into account. In AR, vasodilators reduce afterload mismatch and can preserve LV function and delay the need for surgery. However, if the patient has severely reduced diastolic blood pressure, vasodilators could potentially impair coronary perfusion. In MR, vasodilators may reduce regurgitant volume and LV preload depending on the mechanism of MR. In patients with MR caused by dilated cardiomyopathy, vasodilators reduce symptoms, and improve functional class. However, in mitral valve prolapse or hypertrophic cardiomyopathy, vasodilators may worsen the MR and should be avoided. In other primary causes of MR, vasodilators could potentially mask the development of LV dysfunction and lead to unnecessary and harmful delays in surgery. PMID- 11014376 TI - Focal bacterial nephritis (lobar nephronia) presenting as renal mass. AB - A focal infection of the kidney can cause a diagnostic dilemma by mimicking a neoplasm. We describe a case of focal bacterial nephritis (acute lobar nephronia) caused by Escherichia coli in which the diagnosis was confirmed only after surgical exploration. Although the patient had fever on admission, urine and blood cultures were negative and fine needle aspiration of the kidney could not rule out a well-differentiated carcinoma. PMID- 11014377 TI - Hereditary angioedema precipitated by estrogen replacement therapy in a menopausal woman. AB - We report the first documented case in the literature of hereditary angioedema presenting after commencement of estrogen replacement therapy for menopausal symptoms. The late presentation of the disease and the precipitation of attacks by physiological doses of estrogen replacement therapy make this a highly unusual case. The pathophysiology of hereditary angioedema and its hormonal links are discussed. PMID- 11014378 TI - A subtherapeutic international normalized ratio despite increasing doses of warfarin: could this be malabsorption? AB - OBJECTIVE: To describe a case of warfarin resistance apparently caused by malabsorption and to review the literature regarding warfarin resistance. CASE SUMMARY: A 28-year-old renal transplant patient with systemic lupus erythematosus was admitted for upper extremity thrombophlebitis. Resistance to oral warfarin was demonstrated. Potential causes were investigated. The trapezoidal rule was used to compare the area under the curve for intravenous versus oral dosing of warfarin. The usual bioavailability of warfarin should be 100%. In this patient, warfarin bioavailability after oral dosing was 1.5%. Three potential causes, malabsorption (FF), enzymatic degradation (FG), and first-pass extraction in the portal circulation (FH), are discussed. CONCLUSION: This case demonstrates resistance to warfarin presumably caused by malabsorption. PMID- 11014379 TI - Respiratory bronchiolitis: an unusual cause of pulmonary infiltrates in a pregnant woman. AB - Numerous mechanical, biochemical, and immunologic changes occur during pregnancy. Because of these changes, pregnant women are more susceptible to infection, thromboembolic disease, exacerbation of underlying immunologic disease, and heart failure than women who are not pregnant. The differential diagnosis of diffuse pulmonary infiltrates in a pregnant woman is broad; thus, the work-up can be very challenging. If the patient fails to respond to conservative measures, such as antibiotics, the cause of the infiltrate must be aggressively evaluated because the treatment may be essential for the survival of the mother and fetus. We report a case of a pregnant woman who presented with diffuse bilateral infiltrates. After video-assisted thoracoscopic lung biopsy, this patient was found to have respiratory bronchiolitis, a disease not previously reported during pregnancy. Treatment with glucocorticoids resulted in a prompt improvement in symptoms. PMID- 11014380 TI - Nitric oxide decreases insulin resistance induced by high-fructose feeding. AB - The effect of nitric oxide (NO) on insulin resistance was studied in high fructose-fed rats. A sequential hyperinsulinemic euglycemic clamp procedure was employed (insulin infusion rates: 3 and 30 mU/kg BW/min) in 12 high-fructose-fed rats and 12 chow-fed rats while awake. Half of the high-fructose-fed and the chow fed rats, respectively, were continuously given sodium nitroprusside (SNP, 3 ng/kg BW/min) during the clamp study. Blood glucose was clamped at the fasting level in each rat. Plasma insulin levels during the 3 and 30 mU/kg BW/min insulin infusions were 30 and 400 microU/ml, respectively. Metabolic clearance rate of glucose (MCR) was regarded as an index of whole body insulin action. At both 3 and 30 mU/kg BW/min insulin infusions, high-fructose feeding showed a significant decrease in MCR compared with the chow-fed rats. However, decreased MCRs were stimulated by SNP administration and reached similar levels as the chow-fed rats. SNP infusion did not influence MCRs in the chow-fed rats. Therefore it could be concluded that NO can improve insulin resistance induced by high-fructose feeding. PMID- 11014381 TI - Expression of apolipoprotein B-100 in isolated human small intestine epithelium. AB - Apolipoprotein B-48 (apoB48) is synthesized in the small intestine and becomes a component of chylomicrons (CM). Apolipoprotein B-100 (apoB100) is synthesized in liver and becomes a component of both very low density lipoprotein (VLDL) and low density lipoprotein (LDL). To evaluate whether apoB100 is present in the human small intestine, we performed immunohistochemical staining using anti-apoB100 monoclonal antibody (mAb). Jejunal samples stained positive and the granular staining was noted in the supranuclear region of epithelial cells. We also identified apoB100 expression in the epithelial cells by immunoblotting and dot blotting of PCR-amplified cDNA. In order to exclude submucosal stroma contaminated with blood, we used isolated epithelium from human small intestine obtained by a crypt isolation technique. The results indicate that not only apoB48, but also apoB100 are expressed in human small intestine epithelium. The expression of apoB100 suggests that the dietary VLDL may be synthesized in human small intestine epithelium and converted into LDL, which might play an important role in atherosclerosis. PMID- 11014382 TI - Interleukin-1 potentiates basal and AVP-stimulated ACTH secretion in vitro--the role of CRH pre-incubation. AB - The acute-phase cytokine interleukin-1 (IL-1) is known to activate the hypothalamic pituitary adrenal axis, primarily via corticotropin releasing hormone (CRH). The aim of this study was to determine whether IL-1beta could directly stimulate ACTH secretion from perifused equine anterior pituitary cells, and whether CRH pre-incubation affected corticotroph responsiveness. Isolated equine anterior pituitary cells were pre-incubated with media containing 10 nM CRH or vehicle for 20 hours before being loaded onto columns and perifused with 0.02 nM CRH and 100 nM cortisol. Columns were given a 5-minute pulse of arginine vasopressin (AVP, 10 nM), perifused for 4 hours with 0 (control) or 1 nM IL 1beta, then given a further 5-minute pulse of AVP (10nM). ACTH was measured in 5 minute fractions. In the setting of CRH pre-incubation, cells perifused with IL 1beta for 4 hours showed increased basal ACTH secretion compared to control (114 +/- 6 pM vs. 86 +/- 4 pM [means +/- S.E.M.], p < 0.001) and a significantly greater ACTH response to the final AVP pulse (240 +/- 32% vs. 96 +/- 30%, p = 0.009, expressed as % of ACTH response to the initial AVP pulse). The potentiation of AVP-stimulated ACTH release by IL-1 was not observed in cells pre incubated with vehicle alone. In conclusion, IL-1 increases ACTH release in equine corticotroph cells pre-incubated with CRH and potentiates responsivity to AVP. PMID- 11014383 TI - Association between parathyroid hormone (PTH)/PTH-related peptide receptor gene polymorphism and the extent of bone mass reduction in primary hyperparathyroidism. AB - Genetic contributions to bone mineral density (BMD) and bone turnover are well known. In the present study, we analyzed the relationship between polymorphism of parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor gene existing in exon M7 and the clinical characteristics of primary hyperparathyroidism (pHPT). PTH/PTHrP receptor genotypes were analyzed in 92 pHPT patients by direct sequence to determine whether nucleotide 1417 of the cDNA was C or T. BMD levels at the lumbar spine and at the radius before and one year after parathyroidectomy, as well as serum levels of calcium, phosphorus, alkaline phosphatase (ALP) and intact PTH were measured. Although there were no significant differences in serum levels of calcium, phosphorus and intact PTH, ALP was significantly lower in the CT genotype compared with the TT genotype. BMD level at the radius was significantly higher in the CT genotype than in the CC genotype. Moreover, an increase in radial BMD one year after parathyroidectomy was significantly less in CT genotype than two other genotypes (CC, TT). The present study is the first to indicate that the polymorphism of PTH/PTHrP receptor gene is closely related to the extent of bone mass reduction in pHPT and that this polymorphism would be one of the genetic factors responsible for the severity of the pathological state of pHPT. PMID- 11014384 TI - Effect of subclinical hypothyroidism on body fluid compartments. AB - OBJECTIVE: The present study was aimed to assess the effects of subclinical hypothyroidism on body composition (BC). SUBJECTS: Thirty-one women (age: 37 +/- 9.9 years) with a wide range of body mass index (BMI) were studied. Subclinical hypothyroidism was defined by a basal TSH > or = 4 mU/L and/or TRH stimulated peak > or = 30 mU/L. MEASUREMENTS: For each subject, weight, height, BMI, multifrequency bioelectrical impedance spectroscopy (BIS) and D2O and NaBr dilution tests were performed to assessed total body water (TBW) and extracellular water (ECW). Thyroid function (basal and TRH stimulated TSH, free T3, and free T4) were determined from fasting blood samples for all subjects. Total body dual energy X-ray absorptiometry (DXA) were used to measure fat mass (FM) and lean mass (Lean). RESULTS: The results of BIS were compared with the TBW and ECW estimated by the dilution techniques on the same individuals. The correlation was R2 = 0.65 for impedance at 5 kHz and ECW by NaBr and R2 = 0.72 for impedance at 100 kHz and TBW by D2O. Intracellular water (ICW) was calculated as differences between TBW and ECW measured by dilution methods. Percent of ECW and ICW were related to BMI (ANOVA, p < 0.001). No difference in TBW, body water distribution and body composition related to thyroid function was demonstrated. CONCLUSIONS: In our patients affected with subclinical hypothyroidism, with or without obesity, only obesity appeared related to TBW, ECW and ICW; the subclinical hypothyroidism, on the contrary, had no effect on compartments of body fluids. Bioimpedance is a valid tool to assess body fluid distribution in subclinical hypothyroidism. PMID- 11014385 TI - Myxedema coma of both primary and secondary origin, with non-classic presentation and extremely elevated creatine kinase. AB - Myxedema coma is a rare, often fatal endocrine emergency that concerns elderly patients with long-standing primary hypothyroidism; myxedema coma of central origin is exceedingly rare. Here, we report a 37-year-old woman in whom classical symptoms of hypothyroidism had been absent. Six years earlier, she had severe obstetric hemorrhage and, shortly after, two subsequent episodes of pericardial effusion. On the day of admission, pericardiocentesis was performed for the third episode of pericardial effusion. Because of the subsequent grave arrhythmias and unconsciousness, she was transferred to our ICU. Prior to the endocrine consultation, a silent myocardial infarction had been suspected, based on the extremely high serum levels of creatine kinase (CK) and isoenzyme CK-MB. However, based on thyroid sonography, pituitary computed tomography, elevated titers of antithyroid antibodies and pituitary stimulation tests, the final diagnosis was myxedema coma of dual origin: an atrophic variant of Hashimoto's thyroiditis and post-necrotic pituitary atrophy (Sheehan syndrome). Substitutive therapy caused a prompt clinical amelioration and normalization of CK levels. Our patient is the first case of myxedema coma of double etiology, and illustrates how its presentation deviates markedly from the one endocrinologists and physicians at ICU are prepared to encounter. In addition, cardiac problems as those of our patient should not discourage from substitutive treatment (using L-thyroxine and the gastrointestinal route of absorption), if the age is relatively low. PMID- 11014386 TI - HIV-1 protease inhibitors induce an increase of triglyceride level in HIV infected men without modification of insulin sensitivity: a longitudinal study. AB - We investigated longitudinally the effect of protease inhibitors (PI) on insulin sensitivity, glycemia, and serum lipids in HIV-infected patients. Ninety-one consecutive patients treated with PI for at least 12 months were included in this study. Fasting glycemia, lipid profile, insulinemia, CD4 T lymphocytes, and plasma HIV-1 RNA were performed at baseline and on PI therapy. Insulin sensitivity and insulin secretion were measured by the homeostasis model assessment (HOMA MODEL) using the fasting glucose and insulin concentrations. Triglycerides (+ 0.34 mmol/l, SD = 1.07, p = 0.001) and cholesterol (+ 1.07 mmol/l, SD = 1.21, p= 0.001) significantly increased on PI therapy. Fasting glycemia, insulin sensitivity, and insulin secretion were not modified after PI therapy. PI therapy significantly increased body mass index (0.35 kg/m2, p < 0.05). Serum lipid changes correlated with changes in the CD4+ cell count. Lipodystrophy was observed in 40.6% of patients treated with PI. Our longitudinal study found that PI therapy had no major impact on fasting glycemia, insulin sensitivity, and insulin secretion. These findings are not consistent with previous cross-sectional studies, which did not include baseline measurements before PI initiation. However, we observed a similar profile of lipid changes induced by PI therapy. These results suggest that PI could be responsible for the development of hypertriglyceridemia by a mechanism independent of insulin resistance which remains to be elucidated. PMID- 11014387 TI - A novel dominant-negative mutation of the hepatocyte nuclear factor-1alpha gene in Japanese early-onset type 2 diabetes. AB - We investigated the presence and the function of hepatocyte nuclear factor-1alpha (HNF-1alpha) mutations in 26 Japanese subjects with type 2 diabetes. The subjects were between 20 and 39 years of age on diagnosis and had diabetic first-degree relatives. Two different frameshift mutations were found in 2 subjects (8 %). One novel mutation, T539fsdelC (deletion of C in codon 539 for Thr), is predicted to generate a protein of normal 539 residues at the N-terminus followed by an abnormal 119 amino acid protein. The mutation, P291fsinsC (insertion of C in codon 291 for Pro) should lead to production of a truncated protein of 315 amino acids. Transfection reporter assay using MIN6 and HepG2 cells revealed both mutations to have null function in the transactivation of reporter gene expression. When transfected with wild-type gene, these mutations behaved as dominant-negative regulators in both cells. An equimolar amount of T539fsdelC reduced wild-type activity by approximately 80% in MIN6 cells, while the same concentration of P291fsinsc reduced it by 30%. The sequences responsible for the transactivation activity of HNF-1alpha are confined largely to amino acids 547 628, so that the T539fsdelC mutation, which affects this entire region, replacing amino acids 540-631 with an abnormal 119 amino acid protein, may acquire a potent dominant-negative function. PMID- 11014388 TI - Use of an intact mouse skeletal muscle preparation for endocrine vascular studies: evaluation of the model. PMID- 11014389 TI - "Statistical significance and clinical significance are not synonyms!". PMID- 11014390 TI - Amitriptyline and fluphenazine in the treatment of postherpetic neuralgia. AB - BACKGROUND: Postherpetic neuralgia (PHN) is a vexing problem occurring in 10 to 20 percent of people with from herpes zoster (shingles). Anecdotal reports show that fluphenazine enhances the effects of amitriptyline for the treatment of PHN. The aim of this study was to determine, in a controlled manner, whether this was the case. METHODS: In a double-blind placebo-controlled study, 49 patients with PHN were randomly assigned to four treatment groups: Group 1, amitriptyline; Group 2, amitriptyline and fluphenazine; Group 3, fluphenazine; Group 4, a placebo. An active placebo was used to mimic the anticholinergic side effects of dry mouth. The study lasted 8 weeks, with weekly progress evaluations with use of visual analog scales (VAS), the McGill Pain Questionnaire (MPQ), and a side effects scale. RESULTS: A statistically significant decrease was seen in pain in Groups 1 and 2, and no significant changes were seen in Groups 3 and 4. There was no significant difference when fluphenazine was added to amitriptyline. CONCLUSION: These data support the effectiveness of amitriptyline in treatment of PHN, but do not support the addition of fluphenazine. PMID- 11014391 TI - Flumazenil potentiation of postoperative morphine analgesia. AB - OBJECTIVE: The goal of this study was to test the effect of concomitant administration of flumazenil (FL) and morphine (MO) on immediate postoperative analgesia and the MO requirement to control pain in human beings. DESIGN AND INTERVENTIONS: Thirty-six patients undergoing inguinal hernioplasty under lidocaine epidural anesthesia were enrolled in this double-blind, randomized, controlled study. On the first complaint of pain, either MO (2 mg) only or MO (2 mg) plus FL (0.2 mg) was administered. Additional doses of the same medications administered via a patient-controlled analgesia device with a 10-minute lockout period were available thereafter. The study continued for 2 hours after the loading doses of the medications were administered, with an additional 2-hour period of observation. RESULTS: Thirty-two patients completed the study. Both groups reached a similar satisfactory equianalgesic state (2 in a 0-10 visual analogue scale). The MO plus FL group consumed 9.5 +/- 1.1 mg of MO versus 14.1 +/- 1.1 mg of MO (p < 0.001) in the MO only group. The MO plus FL patients were subjectively (visual analogue scale) more comfortable and less sedated than the MO patients. "Fine" coordination (using an electronic maze) and "coarse" coordination (measured by transferring a pen from one hand to another as rapidly as possible with both arms placed inside an 80-cm metal frame) in the MO group were worse than in the MO plus FL group. End-tidal CO2 increased and blood pressure decreased in the MO group. There were few and insignificant side effects in the MO group. None of these patients required an MO antagonist, and recovery was prolonged in none. CONCLUSIONS: Flumazenil afforded lower MO consumption during the immediate postoperative period. Cognitive, hemodynamic, and respiratory functions were better after MO plus FL than after MO alone. PMID- 11014392 TI - Evaluating skin anesthesia after administration of a local anesthetic system consisting of an S-Caine patch and a controlled heat-aided drug delivery (CHADD) patch in volunteers. AB - OBJECTIVE: The objective of this study was to evaluate the depth and duration of skin anesthesia after the administration of a local anesthetic system consisting of an S-Caine (Zars, Salt Lake City, UT) patch coupled with a controlled heat aided drug delivery (CHADD; Zars) patch. DESIGN: The study design was a randomized, double-blind, placebo-controlled, two-period crossover trial. PATIENTS: Twelve healthy adult volunteers between the ages of 18 and 50 years were enrolled. INTERVENTIONS AND OUTCOME MEASURES: After administration of the study drug or placebo, vital signs (blood pressure, pulse, respiratory rate) were monitored and recorded, and depth and duration of anesthesia were determined and recorded at defined intervals for 10 to 120 minutes after treatment. Depth of anesthesia was measured with a 21-gauge short-bevel needle attached to a depth gauge, and duration was measured using a 0 to 2 (0 = no sensation, 1 = dull sensation, 2 = sharp scratching sensation) verbal report scale. RESULTS: Statistically significant differences were noted in both depth and duration of anesthesia between the active and placebo groups. The posttreatment mean for anesthetic depth in the active group was 6.8 mm compared with 4.7 mm for control group (p = 0.050). The median anesthetic duration was greater than 120 minutes for the active group compared with less than 10 minutes for the placebo group (p = 0.001). CONCLUSIONS: The local anesthetic system consisting of a combination of S-Caine and CHADD patches provided a statistically significant dermal anesthesia effect compared with placebo in this volunteer study. If confirmed in other studies, this system has promise as a noninvasive method of producing dermal anesthesia for minor surgical procedures or intravenous insertion. PMID- 11014393 TI - Topical lidocaine patch relieves a variety of neuropathic pain conditions: an open-label study. AB - OBJECTIVE: Our goal was to perform a pilot study to assess the effectiveness and tolerability of a topical lidocaine patch (Lidoderm) for the treatment of peripheral neuropathic pain conditions other than postherpetic neuralgia. DESIGN: This was an open-label prospective study. PATIENTS: Sixteen patients with refractory peripheral neuropathic pain conditions who had reported intolerable side effects or inadequate pain relief with antidepressant, anticonvulsant, antiarrhythmic, and opioid medications participated in this study. Diagnoses included postthoracotomy pain, stump neuroma pain, intercostal neuralgia, diabetic polyneuropathy, meralgia paresthetica, complex regional pain syndrome, radiculopathy, and postmastectomy pain. OUTCOME MEASURES: A six-item Pain Relief Scale was used (0 = worse pain, 1 = no change, 2 = slight relief, 3 = moderate relief, 4 = a lot of relief, 5 = complete relief). RESULTS: Moderate or better pain relief was reported by 13 of the 16 participants (81%). One patient stopped treatment after 4 days due to lack of relief. The remaining 15 patients had a mean duration of patch use of 6.2 weeks with continued relief. Only 1 patient reported a side effect, a mild skin irritation. CONCLUSIONS: The Lidoderm patch provided clinically meaningful pain relief in most of these refractory neuropathic pain patients without side effects. Controlled trials need to be performed to confirm these preliminary findings. PMID- 11014394 TI - Fitness and perceived exertion in patients with fibromyalgia syndrome. AB - OBJECTIVE: The aim of this study was to evaluate the cardiorespiratory fitness and perceived exertion of female patients with fibromyalgia syndrome (FMS) compared with that of healthy female subjects. DESIGN AND SUBJECTS: This was designed as a cross-sectional case-control study, with a consecutive sample of 30 female patients with FMS and an age-matched control group of 67 healthy female subjects. SETTING: This study was conducted at the multidisciplinary pain center of a university hospital in a city of more than 1 million inhabitants. OUTCOME MEASURES: A cardiorespiratory fitness index (PWC65%/kg) and an original perceived exertion index (B65%) were obtained from the heart rates and perceived exertions scored on a 10-point Borg scale during a submaximal cycle ergometer test. Average indexes for the FMS patients and control subjects were compared. RESULTS: The mean cardiorespiratory fitness index of the FMS patients was not significantly different from that of the controls. The mean perceived exertion index in the FMS patients was significantly greater than that of the controls, meaning that the FMS patients systematically reported higher ratings of perceived exertion during exercise. CONCLUSIONS: Cardiorespiratory fitness, as expressed by a submaximal work capacity index, seems normal in female patients with FMS compared with age- and sex-matched healthy individuals. The fact that FMS patients overscore their perception of exertion may be due to a greater overlap of peripheral pain and perceived exertion perceptions during exercise. This observation should be noted when using perceived exertion scores to prescribe and monitor exercise in FMS patients. PMID- 11014395 TI - Biopsychosocial screening questionnaire for patients with low back pain: preliminary report of utility in physiotherapy practice in Northern Ireland. AB - OBJECTIVE: The aim of this study was to investigate the potential utility of a new biopsychosocial screening questionnaire (Acute Low Back Pain Screening Questionnaire) by exploring the relation between it and several physical risk factors and posttreatment outcomes so as to establish a cutoff point for the local population. The relation between the screening questionnaire and valid and reliable outcome measures of pain and functional disability was also explored. DESIGN: Cross-sectional and longitudinal studies were conducted on patients referred for physiotherapy for low back pain to a large Healthcare Trust in Northern Ireland. Before initial assessment, patients completed the screening questionnaire and outcome measures, were questioned about known physical risk factors, and then received physiotherapy. At final discharge, the outcome variables--the "number of treatments" and patient's current work status ("return to work [yes/no]")--were recorded, and patients recompleted the outcome measures. PATIENTS: One hundred eighteen patients gave written informed consent to participate in this study. RESULTS: Significant associations were detected between questionnaire scores and pretreatment "leisure time exercise," "analgesic medication use," and "subjective anesthesia"; posttreatment "return to work"; and "number of physiotherapy treatments" as well as pain and functional disability measures. A cutoff "at-risk" score of 112 was calculated, which correctly classified 74% of patients who received more than six treatments and 80% of patients who failed to return to work at the end of treatment. CONCLUSIONS: The findings of this study provide preliminary evidence of the utility of this biopsychosocial screening questionnaire for future use in clinical intervention studies in the Northern Ireland National Health Service. Further comparative investigations in other health care settings are warranted. PMID- 11014396 TI - Chronic pain among children and adolescents: physician consultation and medication use. AB - OBJECTIVE: The goal of this study was to assess physician consultation and use of medication in Dutch children and adolescents (0-18 years old) having chronic pain in relation to sociodemographic factors and pain characteristics. DESIGN: This was a population-based cross-sectional survey. A questionnaire was either mailed to the participants' parents or distributed at school, and it was filled out by the parents (for children aged 0-7 years) or by the participant (for children and adolescents aged 8-18 years). SETTING: The study was conducted in the Rotterdam area. PATIENTS: Participants included a random sample of 1,300 children aged 0 to 3 years taken from the register of population. In addition, 41 schools were selected to obtain a representative sample of 5,336 children and adolescents aged 4 to 18 years. OUTCOME MEASURES: Reported physician consultation and medication use were assessed. RESULTS: Of the 6,636 children and adolescents surveyed, 5,424 (82%) responded. A total of 1,358 respondents (25%) reported chronic pain. Of these, 57% had consulted a physician and 39% had used medication for the pain. Respondents with earache, more intense pain, and more frequent pain and those attending lower vocational training programs were more likely to consult a physician for the pain than the average respondent. Respondents with earache, sore throat, headache, more intense pain, and multiple pain; children aged 0 to 3 years; and girls were more likely to use medication for the pain. Logistic regression analyses showed that for physician consultation, the most significant predictive factors were the intensity of pain, age, and earache as well as the level of education for respondents aged 12 to 16 years. The use of medication was predicted by earache, headache, limb pain, intensity of pain, and age. CONCLUSIONS: Chronic pain is a common complaint in children and adolescents, frequently resulting in consultation of a physician and medication use. Regarding physician consultation, children and adolescents with a lower educational level seem to be a group at risk. PMID- 11014397 TI - Predictors of participation in primary care group-format back pain self-care interventions. AB - OBJECTIVE: The goal of this study was to explain how primary care back pain patients who volunteer for a group-format self-care intervention differ from nonvolunteers. This is relevant to the generalizability of studies that rely on volunteers as well as the characteristics of patients who do not seek out self care interventions. SETTING: This study was conducted at a large health maintenance organization in western Washington state. PATIENTS: "Volunteers" (n = 481) were primary care back pain patients participating in randomized trials of a self-management intervention who were recruited through passive nonintensive means (a mailed invitation). "Nonvolunteers" (n = 967) consisted of a representative sample of consecutive back pain patients. We compared the baseline characteristics of these two groups. RESULTS: The relatively small percentage (8%) of primary care back pain patients who volunteered for, and ultimately participated in, group self-management classes tended to be white, older, better educated, and more likely to be retired than nonvolunteers. The two groups did not differ significantly on most clinical measures, including pain intensity and persistence. Patients experiencing the highest (and lowest) levels of pain related activity interference were less likely to volunteer than those with moderate activity limitations, however. CONCLUSIONS: Those individuals volunteering to participate in a group-format self-care intervention in a primary care setting differed from nonvolunteers primarily on demographic measures as opposed to clinical measures. Back pain patients experiencing the highest levels of activity limitations were somewhat less likely to participate than those with moderate activity limitations. Recruitment for effective self-care interventions is an important issue in determining their impact on a population basis. PMID- 11014398 TI - Use of virtual reality for adjunctive treatment of adult burn pain during physical therapy: a controlled study. AB - OBJECTIVE: The pain experienced by burn patients during physical therapy range of motion exercises can be extreme and can discourage patients from complying with their physical therapy. We explored the novel use of immersive virtual reality (VR) to distract patients from pain during physical therapy. SETTING: This study was conducted at the burn care unit of a regional trauma center. PATIENTS: Twelve patients aged 19 to 47 years (average of 21% total body surface area burned) performed range of motion exercises of their injured extremity under an occupational therapist's direction. INTERVENTION: Each patient spent 3 minutes of physical therapy with no distraction and 3 minutes of physical therapy in VR (condition order randomized and counter-balanced). OUTCOME MEASURES: Five visual analogue scale pain scores for each treatment condition served as the dependent variables. RESULTS: All patients reported less pain when distracted with VR, and the magnitude of pain reduction by VR was statistically significant (e.g., time spent thinking about pain during physical therapy dropped from 60 to 14 mm on a 100-mm scale). The results of this study may be examined in more detail at www.hitL.washington.edu/projects/burn/. CONCLUSIONS: Results provided preliminary evidence that VR can function as a strong nonpharmacologic pain reduction technique for adult burn patients during physical therapy and potentially for other painful procedures or pain populations. PMID- 11014399 TI - Hypogonadism in patients treated with intrathecal morphine. AB - OBJECTIVE: The objective of this study was to investigate the hypothalamic pituitary-gonadal response to intrathecal opioids. PATIENTS: Thirty patients receiving intrathecal morphine for chronic nonmalignant pain were studied for clinical and biochemical evidence of hypogonadism. Ten men and 10 postmenopausal women with chronic pain of similar duration but who were not receiving any form of opioid therapy acted as control subjects. RESULTS: Men and both premenopausal and postmenopausal women had evidence of hypogonadism with low levels of serum testosterone or estrogen coupled with low levels of pituitary gonadotrophins. Control subjects had hormone levels in the expected range for their sex and age. Two men demonstrated recovery after ceasing intrathecal opioid therapy. CONCLUSIONS: Hypogonadotrophic hypogonadism is a common complication of intrathecal opioid therapy in both men and women. PMID- 11014400 TI - Effect of acupuncture upon experimentally induced ischemic pain: a sham controlled single-blind study. AB - OBJECTIVE: To investigate the hypoalgesic effect of true and sham acupuncture upon experimentally induced ischemic pain. DESIGN: Human volunteers (n = 60) were required to attend two sessions for pain induction using a submaximal effort tourniquet technique; on the first occasion, baseline pain scores were recorded and on the second, 48 hours later, subjects were randomly allocated to one of five groups: Control, Treatment Groups 1 or 2, or Placebo Groups 1 or 2. SUBJECTS: Healthy human volunteers. INTERVENTION: In all the Treatment and Placebo Groups, subjects received some form of needle acupuncture 15 minutes before, and 5 minutes during, the pain induction procedure on the second day. Treatment Group 1 received acupuncture on acupuncture points situated distal to the tourniquet, whereas Treatment Group 2 received acupuncture on acupuncture points situated proximal to the tourniquet. In Placebo Groups 1 and 2, subjects received 'sham' acupuncture either on nonacupuncture points (Placebo Group 1) or on acupuncture points (Placebo Group 2) using (standardized) minimal levels of stimulation. A licensed acupuncturist who was not involved in data collection and analyses carried out all treatments. OUTCOME MEASURES: Pain was assessed using a computerized visual analog scale (VAS) and a McGill Pain Questionnaire (MPQ). RESULTS: Analysis of VAS scores using ANOVA revealed no significant differences between groups (e.g., VAS sum of differences data (mean +/- SEM): Treatment Group 1: 90+/-47, Treatment Group 2: 187+/-56, Placebo Group 1: 152+/-40, Placebo Group 2: 121+/-42, CONTROLS: 46+/-24, p>0.05). Analysis of MPQ percentage difference scores using one-way ANOVA revealed some isolated effects in the subjective descriptors and the Pain Rating Index, both for Treatment Group 2 and Placebo Group 2, proving them superior to any of the other groups. CONCLUSION: The results of the study provide no convincing evidence for a superior hypoalgesic effect of acupuncture compared with "sham" procedures on this model of experimental pain. PMID- 11014401 TI - Severe lightning pain after subarachnoid block in a patient with neuropathic pain of central origin: which drug is best to treat the pain? AB - OBJECTIVE: There have been many reports that spinal anesthesia induces severe lightning pain in the lower limbs of patients with phantom limb pain, tabes dorsalis, or causalgia. We report on a patient with neuropathic pain of central origin who showed newly developed severe lightning pain after therapeutic subarachnoid block (SAB). We performed SAB 16 times in this patient, and he complained of severe pain each time. We investigated which drug was best for treating such induced pain by administering various drugs to the patient. SETTING: The patient was hospitalized for treatment and observation. PATIENT: The patient was a 48-year-old man with neuropathic pain secondary to an incomplete spinal cord injury at the cervical segment. INTERVENTIONS: Various drugs were administered for relieving the newly developed severe pain, and the effectiveness of these agents was compared. RESULTS AND CONCLUSIONS: Intravenous thiopental, fentanyl, butorphanol, ketamine, midazolam, droperidol, and sevoflurane-oxygen anesthesia were quite effective. Intramuscular butorphanol was not effective. Intravenous physiologic saline and atropine sulfate as a placebo, intrathecal morphine hydrochloride, intravenous mexiletine, and lidocaine were ineffective. Intravenous thiopental (approximately 1 mg/kg) was thought to obtain the best pain relief because it stopped the pain quickly, the dose needed was subanesthetic, and there was no adverse effect. PMID- 11014402 TI - Sex-specific and general roles of pain-related anxiety in adjustment to chronic pain: a reply to Edwards et al. PMID- 11014403 TI - Phage therapy: the peculiar kinetics of self-replicating pharmaceuticals. AB - The specter of antibiotic-resistant bacteria has provoked renewed interest in the possible use of bacteriophages to control bacterial infections. We argue that clinical application of phage therapy has been held back by a failure to appreciate the extent to which the pharmacokinetics of self-replicating agents differ from those of normal drugs. For self-replicating pharmaceutical agents, treatment outcome depends critically on various density-dependent thresholds, often with apparently paradoxical consequences. An ability to predict these thresholds and associated critical time points is a necessity if phage therapy is to become clinically practicable. PMID- 11014404 TI - Increased drug delivery to the brain by P-glycoprotein inhibition. AB - BACKGROUND: Although the antidiarrheal loperamide is a potent opiate, it does not produce opioid central nervous system effects at usual doses in patients. On the basis of in vitro studies demonstrating that loperamide is a substrate for the adenosine triphosphate-dependent efflux membrane transporter P-glycoprotein, we postulated that inhibition of P-glycoprotein with quinidine would increase entry of loperamide into the central nervous system with resultant respiratory depression. METHODS: To test this hypothesis, a 16-mg dose of loperamide was administered to eight healthy male volunteers in the presence of either 600 mg quinidine, a known inhibitor of P-glycoprotein, or placebo. Central nervous system effects were measured by evaluation of the respiratory response to carbon dioxide rebreathing as a measure of opiate-induced respiratory depression. RESULTS: Loperamide produced no respiratory depression when administered alone, but respiratory depression occurred when loperamide (16 mg) was given with quinidine at a dose of 600 mg (P < .001). These changes were not explained by increased plasma loperamide concentrations. CONCLUSION: This study therefore demonstrates first the potential for important drug interactions to occur by a new mechanism, namely, inhibition of P-glycoprotein, and second that the lack of respiratory depression produced by loperamide, which allows it to be safely used therapeutically, can be reversed by a drug causing P-glycoprotein inhibition, resulting in serious toxic and abuse potential. PMID- 11014405 TI - Pharmacokinetics of L-carnitine in patients with end-stage renal disease undergoing long-term hemodialysis. AB - OBJECTIVE: L-Carnitine is an endogenous molecule involved in fatty acid metabolism. Secondary carnitine deficiency may develop in patients with end-stage renal disease undergoing long-term hemodialysis because of dialytic loss. In these patients L-carnitine can be administered to restore plasma and tissue levels. The objective of this study was to evaluate the pharmacokinetics of intravenous L-carnitine in patients undergoing long-term hemodialysis. METHODS: Twelve patients undergoing three dialysis sessions/week received L-carnitine intravenously (20 mg x kg(-1)) at the end of each dialysis session for 9 weeks. Plasma samples were analyzed for L-carnitine, acetyl-L-carnitine, and total carnitine by HPLC. RESULTS: Under baseline conditions, the mean +/- SD predialysis plasma concentration of L-carnitine was 19.5 +/- 5.6 micromol/L, decreasing to 5.6 +/- 1.9 micromol/L at the end of the dialysis session. These concentrations were substantially lower than endogenous levels in healthy human beings. Under baseline conditions the extraction ratios of L-carnitine and acetyl L-carnitine by the dialyser were 0.74 +/- 0.07 and 0.71 +/- 0.11, respectively. During repeated dosing, there was accumulation of L-carnitine in plasma, and after 9 weeks of dosing, the predialysis and postdialysis plasma levels were 191 +/- 54.1 and 41.8 +/- 13.0 micromol/L, respectively. The predialysis and postdialysis plasma levels of L-carnitine decreased once dosing was ceased but had not returned to pretreatment levels after 6 weeks. CONCLUSION: The study demonstrated that removal of L-carnitine by hemodialysis is extremely efficient and that patients undergoing hemodialysis had plasma concentrations that were substantially lower than normal, particularly during dialysis. During repeated administration of L-carnitine, the predialysis and postdialysis concentrations of the compound increased steadily, reaching an apparent steady state after about 8 weeks. It is proposed that this accumulation arose from the distribution of L carnitine into a deep tissue pool that includes skeletal muscle. PMID- 11014406 TI - Pharmacokinetics of nicotine in kidney failure. AB - BACKGROUND: Smoking is an important risk factor for cardiovascular and cerebrovascular complications in patients with chronic kidney failure. Very high plasma nicotine concentrations have been reported in patients with severe kidney failure, indicating that the disposition of nicotine in these patients may be different. The purpose of this study was to assess the pharmacokinetics of intravenously administered nicotine in healthy subjects and in patients with kidney failure. METHODS: Nine healthy subjects (glomerular filtration rate [GFR], 84 to 143 mL/min/1.73 m2), four patients with mild kidney failure (GFR, 63 to 73 mL/min/1.73 m2), five patients with moderate kidney failure (GFR, 18 to 36 mL/min/1.73 m2), and six patients with severe kidney failure (GFR, 1 to 10 mL/min/1.73 m2) were recruited. Three patients were treated with continuous ambulatory peritoneal dialysis. An intravenous infusion of nicotine (0.028 mg/kg) was given for 10 minutes. Nicotine and cotinine concentrations were measured in plasma, urine, and peritoneal dialysate from 0 to 24 hours after start of infusion RESULTS: There were significant correlations between GFR and total clearance, nonrenal and renal clearance of nicotine, area under the plasma concentration-time curve extrapolated to infinity, terminal elimination half life, and mean residence time. Nonrenal clearance was 1303 mL/min and 661 mL/min in healthy subjects and patients with severe kidney failure, respectively. Only 1% to 2% of the nicotine dose was excreted unchanged in a 24-hour collection of peritoneal dialysate. The elimination of cotinine was also decreased in patients with kidney failure. CONCLUSION: Progressive kidney failure is associated with a gradual decrease of renal and nonrenal elimination of nicotine. PMID- 11014407 TI - Disposition and safety of omapatrilat in subjects with renal impairment. AB - BACKGROUND: Omapatrilat, a vasopeptidase inhibitor, preserves natriuretic peptides and inhibits the renin angiotensin aldosterone system by simultaneously inhibiting neutral endopeptidase and angiotensin-converting enzyme. METHODS: Oral omapatrilat, 10 mg/d, was administered for 8 to 9 days to three groups of eight subjects with varying degrees of renal function (CLCR values, normal > or = 80; mild to moderate impairment < 80 to > or = 30; severe impairment < 30 mL/min/1.73 mL2) and to six subjects undergoing maintenance hemodialysis. Omapatrilat and its metabolites (phenylmercaptopropionic acid, S-methylomapatrilat, S methylphenylmercaptopropionic acid, and cyclic S-oxide-omapatrilat) were quantified in plasma by a validated liquid chromatography/mass spectrometry method. The model, Cmax or AUC(0-T) = intercept + slope x CLCR, was tested for a possible linear correlation between Cmax (peak plasma concentrations) or AUC(0-T) (area under plasma concentration versus time curve) and CLCR. RESULTS: For omapatrilat and its inactive metabolites, phenylmercaptopropionic acid, S methylomapatrilat, and S-methylphenylmercaptopropionic acid, the median times to peak plasma concentrations (tmax) were 1.5 to 2, 2 to 3, 2.5 to 3.5, and 7 to 10 hours, respectively, and were independent of renal function. After Cmax attainment, plasma concentrations declined rapidly to about 10% of Cmax values. Cyclic S-oxide-omapatrilat, a potentially active metabolite, was undetectable at all sampling time points. Hemodialysis did not decrease circulating levels of omapatrilat. There was minimal accumulation of omapatrilat and phenylmercaptopropionic acid and moderate accumulation of the S-methylated metabolites. For omapatrilat and S-methylphenylmercaptopropionic acid, neither Cmax nor AUC(0-T) was CLCR dependent. However, AUC(0-T) for phenylmercaptopropionic acid and both the Cmax and AUC(0-T) for S methylomapatrilat were CLCR dependent. CONCLUSIONS: The pharmacokinetics of omapatrilat, the only clinically relevant active compound studied, was independent of CLCR. For patients with reduced renal function, adjusting initial omapatrilat dose is not suggested. Hemodialysis did not significantly contribute to the clearance of omapatrilat. The long-term pharmacodynamic response to omapatrilat will dictate dose-adjustment needs. PMID- 11014408 TI - Dose and time dependencies of 5-fluorouracil pharmacokinetics. AB - OBJECTIVES: The purpose of this study was to examine the interpatient and intrapatient variability of the Michaelis-Menten plasma parameters of 5 fluorouracil administered according to a schedule combining a bolus of 400 mg/m2 followed by 22-hour infusion of 600 mg/m2 for 2 consecutive days. PATIENTS: A pharmacokinetic population approach was used to analyze the data from 21 patients with colorectal cancer. RESULTS: The 5-fluorouracil plasma concentrations versus time were best described by a two-compartment model with nonlinear elimination from the central compartment. The relationships between the pharmacokinetic parameters and patient characteristics were tested. On day 1 the mean values (with interindividual variability as expressed by the coefficient of variation) were 1390 mg x h(-1) (20%), and 5.57 mg x L(-1) (22%) for the maximum rate of elimination, and the half-saturating plasma concentration. The maximum rate of elimination was positively correlated to the body surface area and the percentage of liver involvement by metastatic disease determined by tomodensitometric examination. The model was successfully tested with independent data sets corresponding to other schedules. The analysis of this intrapatient variability showed that the half-saturating plasma concentration increased from day 1 to day 2, especially in the patients with low lymphocyte cell dihydropyrimidine dehydrogenase activity. CONCLUSION: The pharmacokinetic parameters obtained in this study would be useful to predict the 5-fluorouracil plasma concentrations following other schedules of administration of 5-fluorouracil and to study the possible pharmacokinetic interactions between 5-fluorouracil and other drugs. PMID- 11014409 TI - In vivo drug-drug interaction studies--a survey of all new molecular entities approved from 1987 to 1997. AB - Ninety-eight new molecular entities applications approved between 1987 to 1991 (period I) and 193 applications for new molecular entities between 1992 to 1997 (period II) were surveyed for drug-drug interaction studies. In period I (used as a comparator), 32 applications contained drug-drug interaction studies for a total of 117 studies. In period II, 106 applications reported drug-drug interaction studies, and the number of studies per new molecular entity ranged from 0 to 15. Most studies (77%) were performed in healthy subjects, with 44% using crossover designs, 7% using parallel designs, and the remaining using fixed sequence designs. The most common dosing scheme for new molecular entities/interacting drug was multiple dose (47%), whereas single dose/multiple dose was used in 31% of studies, and single dose/single dose was used in 18% of studies. Of the 540 drug-drug interaction studies submitted in period II, 80 (15%) resulted in clinically significant labeling statements. Submissions for new molecular entities to the Center for Drug Evaluation and Research divisions most likely to include drug-drug interaction studies were neuropharmacology, cardiorenal, antiviral, and antiinfective drugs. Some drug classes such as oncology drug products and radioimaging products were least likely to include drug-drug interaction studies in their submissions. We conclude that the use of drug-drug interaction studies in the drug development process has increased between the two periods. PMID- 11014411 TI - Effect of hydrocortisone on phenylephrine--mean arterial pressure dose-response relationship in septic shock. AB - BACKGROUND: Septic shock is characterized by decreased responsiveness to catecholamines. Because endogenous steroids are known to play a role in the modulation of vasomotor tone, the purpose of our study was to investigate the phenylephrine-mean arterial pressure dose-response relationship in patients with septic shock and the effect of a physiological dose of hydrocortisone on it. METHODS: Twelve patients meeting usual criteria for septic shock and 12 age matched control subjects were investigated before and 1 hour after receiving 50 mg intravenous hydrocortisone. Sixteen incremental doses of phenylephrine (microg/kg/min) were infused, and the effects on mean arterial pressure (mm Hg) were recorded. A sigmoid model, E = E0 + [Emax x Dgamma/(ED50gamma + Dgamma)], was fitted to individual data. In this model, E is the predicted effect and D is the dose of phenylephrine infused. E0 represents the basal value of effect (ie, the value of mean arterial pressure without drug), Emax is the maximum theoretical effect, ED50 is the dose of phenylephrine for which an effect of 50% of Emax is observed, and gamma is the Hill coefficient which accounts for the sigmoidicity of the curve. RESULTS: As compared with in control subjects, in patients, E0 was decreased before (58 +/- 8 versus 73 +/- 7 mm Hg) and after (64 +/- 12 versus 82 +/- 10 mm Hg) administration of hydrocortisone (P = .0001 for group), Emax was reduced before (39 +/- 17 versus 84 +/- 18 mm Hg) and after (77 +/- 26 versus 106 +/- 21 mm Hg) administration of hydrocortisone (P = .0001 for group), ED50 was not modified, and gamma was increased before (3.5 +/- 1.8 versus 1.3 +/- 0.3) and after (1.9 +/- 1.1 versus 1.3 +/- 0.3) administration of hydrocortisone (P = .0010 for group). Hydrocortisone similarly increased E0 in both groups (P = .0003 for sequence, P = .2883 for interaction), increased more Emax in patients than in control subjects (P < .0001 for sequence; P = .0280 for interaction), did not change ED50, and decreased y in patients but not in control subjects (P = .0025 for sequence, P = .0025 for interaction). CONCLUSIONS: In patients with septic shock, the Emax of phenylephrine is decreased, whereas its ED50 is not modified, both before and after administration of hydrocortisone. A physiological dose of hydrocortisone tends to normalize the relationship. PMID- 11014410 TI - Faster clearance of sustained release verapamil in men versus women: continuing observations on sex-specific differences after oral administration of verapamil. AB - Pharmacokinetic studies after administration of 120 mg oral sustained- and regular-release racemic verapamil were performed in 13 healthy subjects (seven men, age 74 +/- 4 years [mean +/- SD], weight 69.9 +/- 5.4 kg, and body mass index 24.6 +/- 2.2]; and six women, age 65 +/- 13 years, weight 65 +/- 9.9 kg, and body mass index 25.3 +/- 3). Verapamil was measured by HPLC, concentration versus time data analyzed by noncompartmental models, and statistical analyses performed by ANOVA for repeated measurements. The area under the concentration versus time curve (AUC) after administration of sustained-release verapamil was 48,951 +/- 18,079 ng/mL x min(-1) in women compared with 25,595 +/- 10,245 in men and lower than after administration of regular-release verapamil (63,055 +/- 24,411 for women and 34,686 +/- 25,279 in men; P = .05 for sex-related effect and P < .02 for formulation effect). AUC ratios of norverapamil (N-demethylated metabolite) to verapamil after administration of sustained-release verapamil were 1.43 +/- 0.26 in women compared with 1.74 +/- 0.41 in men and 1.43 +/- 0.26 in women compared with 1.78 +/- 0.37 in men after administration of regular-release verapamil (P = .1 for sex-related effect and P = .9 for formulation effect). Apparent oral clearance was 43 +/- 15 mL/min/kg in women compared with 75 +/- 29 in men after administration of sustained-release verapamil and 35 +/- 16 mL/min/kg in women compared with 65 +/- 31 in men after administration of regular release verapamil (P < .05 for sex-related effect and P < .02 for formulation effect). Apparent oral clearance of both regular- and sustained-release formulations of verapamil was faster in men compared with women in contrast to findings after intravenous administration of verapamil, suggesting that intestinal processes are a factor in sex-specific difference in drug clearance. Greater verapamil and norverapamil bioavailability after administration of regular- compared with sustained-release verapamil also suggests saturable processes at the intestinal level. PMID- 11014412 TI - A comparative pharmacodynamic study of IY-81149 versus omeprazole in patients with gastroesophageal reflux disease. AB - OBJECTIVE: To evaluate and compare the pharmacodynamic effects of IY-81149 and omeprazole on gastric pH in patients with gastroesophageal reflux disease. METHODS: Sixty male and female volunteers with gastroesophageal reflux disease were enrolled in a double-blind, two-way, crossover, dose-ranging study. Subjects were randomized into three groups, with each group comparing the effect of one of three doses of IY-81149 (5, 10, or 20 mg) with 20 mg omeprazole. IY-81149 and omeprazole were administered once daily for 5 days. Continuous 24-hour pH measurements were performed before the first dose (baseline) and after the fifth dose in both periods. Gastric acid suppression was evaluated on the basis of the following parameters: AUC(0-24), median pH in a 24-hour interval (pHmedian), and the percent time in a 24-hour interval in which the gastric pH was greater than 4 (tpH > 4). The truncated AUC parameters AUC(0-8), AUC(8-16), and AUC(16-24) were also calculated. The effects of IY-81149 and omeprazole on gastric pH were compared by use of analyses of covariance. The dose-response relationship for IY 81149 was also evaluated. RESULTS: There were no statistically significant differences between 5 mg IY-81149 and 20 mg omeprazole in terms of AUC(0-24), pHmedian, tpH, 4, AUC(0-8), and AUC(8-16). IY-81149, at 10 mg, produced a significantly greater gastric acid suppression than omeprazole on the basis of the values of AUC(0-24), pHmedian, tpH > 4, AUC(8-16), and AUC(16-24). Administration of 20 mg IY-81149 produced a significantly greater gastric acid suppression on the basis of all parameters. All doses of IY-81149 were more effective than omeprazole during 16 to 24 hours after the dose was administered. CONCLUSIONS: Administration of 10 and 20 mg IY-81149 produced a statistically significantly greater and prolonged suppression of gastric pH than 20 mg omeprazole. PMID- 11014413 TI - Ibuprofen plus caffeine in the treatment of tension-type headache. AB - BACKGROUND: The effectiveness of caffeine as an adjuvant to ibuprofen has been documented in investigations of acute pain. Our objectives were to assess this agent in the treatment of tension-type headache and to establish clinical trial methods capable of assessing this agent in comparison with various tension headache treatments. Stopwatch technology was used for measurement techniques. METHODS: A randomized, double-blind, parallel, multicenter, single-dose, placebo- and active-controlled study included 301 subjects diagnosed with tension-type headache. Treatment groups included ibuprofen and caffeine, ibuprofen alone, caffeine alone, or placebo. Subjects measured onset of relief (both time to first perceptible relief and time to meaningful relief) after taking a single oral dose of their assigned medication. Pain intensity and pain relief were rated over a 6 hour study period. Overall evaluation was made on completion of all other ratings. RESULTS: Ibuprofen and caffeine administered together provided significantly greater analgesic activity than ibuprofen alone, caffeine alone, and placebo. Ibuprofen and caffeine administered together demonstrated significantly shorter times to meaningful improvement in headache relief than ibuprofen or placebo; significantly greater total analgesia than ibuprofen alone, caffeine alone, or placebo; and significantly greater peak relief than ibuprofen alone, caffeine alone, or placebo. Significantly more subjects obtained meaningful headache relief with ibuprofen and caffeine administered together than with ibuprofen alone or placebo. More patients reported complete headache relief with ibuprofen and caffeine administered together than with ibuprofen alone, caffeine alone, or placebo. Ibuprofen and caffeine administered together was rated significantly better by patients than either ibuprofen alone, caffeine alone, or placebo. No subjects ended participation in the study early because of adverse events. CONCLUSIONS: Sensitive methods have been introduced to assess differences in analgesia among over-the-counter analgesic agents in relieving tension-type headache pain. A double-blind study with this method suggests that ibuprofen and caffeine administered together provides greater analgesic effectiveness than either component alone. PMID- 11014414 TI - Effects of the 2-amino-3-hydroxy-5-methyl-4-isoxazole-proprionic acid/kainate antagonist LY293558 on spontaneous and evoked postoperative pain. AB - BACKGROUND: Previous studies suggest that 2-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA)/kainate antagonists reduce experimentally induced pain. There have been no studies of AMPA/kainate antagonists in clinical pain. METHODS: Analgesic efficacy of intravenous LY293558 (0.4 or 1.2 mg/kg) was compared with that of intravenous ketorolac tromethamine (INN, ketorolac; 30 mg) and placebo in a randomized, double-blind, parallel-group study after oral surgery (n = 70). Study drugs were administered at the onset of moderate pain; pain intensity and relief were measured for 240 minutes. RESULTS: High-dose LY293558 and ketorolac tromethamine were superior to placebo (P < .05) for pain evoked by mouth opening and one of several measures of spontaneous pain: SPID240 +/- SEM for pain evoked by mouth opening was highest for ketorolac tromethamine (151 +/- 58), intermediate for high-dose LY293558 (-45 +/- 35), and least for low-dose LY293558 (-151 +/- 39) and placebo (-162 +/- 50). High-dose LY293558 was superior to placebo at individual time points (45 to 240 minutes) for pain evoked by mouth opening but not for spontaneous pain. The spontaneous summed pain intensity difference over 240 minutes (SPID240 +/- SEM) was highest for ketorolac tromethamine (303 +/- 84), intermediate for high-dose LY293558 (-51 +/- 40) and low-dose LY293558 (-96 +/- 45), and least for placebo (-180 +/- 24). LY293558 was well tolerated, with dose-dependent and reversible side effects including hazy vision in 20% of patients and sedation in 15%. CONCLUSIONS: This is the first evidence that an AMPA/kainate antagonist reduces clinical pain. Tests of evoked pain may be more sensitive to certain analgesics than those of spontaneous pain. The evaluation of evoked pain as an outcome measure in analgesic trials may identify potentially useful compounds otherwise missed if only spontaneous pain is evaluated. PMID- 11014415 TI - CYP2C19 genetic mutations in North Indians. AB - OBJECTIVES: To identify defective alleles of CYP2C19 (CYP2C19*2 and *3) in North Indians. METHODS: One hundred extensive metabolizers and 21 poor metabolizers of omeprazole were genotyped with respect to CYP2C19*2 and *3 alleles with polymerase chain reaction-based diagnostic tests. RESULTS: Fifty-two extensive metabolizers and six poor metabolizers were homozygous with the CYP2C19*1/*1 genotype, and 48 extensive metabolizers and six poor metabolizers were heterozygous with the CYP2C19*1/*2 genotype. Nine poor metabolizers were homozygous with the CYP2C19*2/*2 genotype. No extensive or poor metabolizers demonstrated the presence of the CYP2C19*3 allele. CYP2C19*2 could explain 43% (9/21) of the poor metabolizers and 57% (24/42) of the defective alleles in poor metabolizers. Allele frequency of CYP2C19*1 and *2 was 0.7 (95% confidence interval of 0.65 to 0.75) and 0.3 (95% confidence interval of 0.25 to 0.35), respectively. Homozygous extensive metabolizers excreted 7.85 +/- 7.6 micromol 5 hydroxyomeprazole in 8 hours, which was 28% more as compared with heterozygous extensive metabolizers who excreted 5.6 +/- 3.6 micromol 5-hydroxyomeprazole in 8 hours (P < .05). CONCLUSIONS: CYP2C19*2 demonstrated allele frequency of 0.3, whereas CYP2C19*3 was absent in North Indians. Because CYP2C19*2 is not able to explain 57% of poor metabolizers, other mutations (CYP2C19*4 to *8) might be present in North Indians. CYP2C19 demonstrated differential evolution in North Indians because the frequency of CYP2C19*2 was similar to that in Oriental subjects, but that of CYP2C19*3 was similar to that in white subjects. PMID- 11014416 TI - Mechanisms of chloroquine-induced pruritus. PMID- 11014417 TI - Triptans and coronary spasm. PMID- 11014419 TI - Cognitive functioning and cerebrospinal fluid concentrations of neuropeptides for patients with good neurological outcomes after aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Many patients exhibit cognitive disturbances after aneurysmal subarachnoid hemorrhage (SAH). Structural and functional neuroimaging has failed to demonstrate any correlation with these complaints. This study was performed to investigate whether neuropeptide concentrations in cerebrospinal fluid could be related to cognitive disturbances after SAH. METHODS: Lumbar cerebrospinal fluid was obtained, 3 to 6 months after surgery, from 17 patients who experienced good outcomes after aneurysmal SAH. The samples were analyzed for various neuropeptides using radioimmunoassays, and the peptide concentrations were evaluated in relation to scores on standardized neuropsychological tests. RESULTS: The neuropsychological test results were normal for eight individuals, whereas the remaining nine patients exhibited various degrees of cognitive impairment. There was no correlation between the concentrations of arginine vasopressin or neuropeptide Y and test performance. However, significant correlations between cognitive impairment and elevated levels of beta-endorphins (P = 0.02), corticotropin-releasing factor (P = 0.004), and delta sleep-inducing peptide (P = 0.045) were noted. CONCLUSION: Patients with cognitive impairments after aneurysmal SAH exhibited higher cerebrospinal fluid concentrations of endorphins, corticotropin-releasing factor, and delta sleep-inducing peptide than did those with normal capacity. This is probably attributable to diffuse derangement of transmitter release in the brain, resulting from the insult or ensuing complications, although a secondary increase in corticotropin-releasing factor concentrations caused by increased stress during the testing because of reduced cognitive capacity cannot be excluded. PMID- 11014418 TI - The insular lobe: physiopathological and surgical considerations. AB - OBJECTIVE: Surgery of the insula represents a technical challenge, because of the proximity of the internal capsule to the lenticulostriate arteries and the lack of certainty concerning its functionality. Using intraoperative direct cerebral stimulation, combined with neuronavigation, the authors operated on 12 insular gliomas. On the basis of this experience, the physiopathological and surgical implications are discussed. METHODS: A low-grade insular glioma, revealed by seizures, was diagnosed in 12 right-handed patients with a normal neurological status. Preoperative magnetic resonance imaging showed that, according to Yasargil's classification system, three patients harbored Type 3 lesions and nine patients had Type 5 lesions (10 tumors on the right side and 2 on the left dominant side). All patients underwent surgery using direct cerebral stimulation, under general anesthesia in nine patients (motor mapping) and under local anesthesia in three patients (sensorimotor and language mapping). Ultrasonography and/or neuronavigation was used in all cases. Preoperative angio-computed tomographic scanning showed the lenticulostriate arteries in two patients. RESULTS: The internal capsule was systematically detected, and the language areas were identified within the left insula in the awake patients. The lenticulostriate arteries were seen in two patients. Seven patients presented an immediate postoperative deficit; six of them recovered completely within 3 months. Four resections were total, six were subtotal, and two were partial (left insula). CONCLUSION: The use of intraoperative direct cerebral stimulation and neuronavigation allows surgery of the insula with minimization of the risk of sequelae, but its use is still limited with regard to the dominant hemisphere, owing to the essential role of this structure in language. PMID- 11014420 TI - Predicting delayed ischemic deficits after aneurysmal subarachnoid hemorrhage using a transient hyperemic response test of cerebral autoregulation. AB - OBJECTIVE: To assess whether the development of delayed ischemic deficits (DIDs) after aneurysmal subarachnoid hemorrhage can be predicted using transcranial Doppler ultrasonography and the transient hyperemic response test (THRT). METHODS: An increase in the middle cerebral artery peak flow velocity (FV) of more than 9% of baseline values after 5 to 9 seconds of carotid artery compression was defined as a normal THRT result, indicating good autoregulatory reserve. The transcranial Doppler criteria for vasospasm were a FV of more than 120 cm/s and a Lindegaard ratio of more than 3. Twenty patients with no immediate postoperative neurological deficits were studied. The FVs at all of the major cerebral arteries were measured daily after surgery, and the THRT results were assessed bilaterally. RESULTS: Five of six patients with abnormal THRT results in the first examination after surgery (primary THRT impairment) developed DIDs; none of the remaining patients developed DIDs (Fisher exact test, P = 0.0004). All five patients with DIDs initially exhibited low FVs but all subsequently developed increases in FVs to values of more than 150 cm/s and four exhibited FVs of more than 200 cm/s. The time of onset of DIDs corresponded to the time of onset of moderate vasospasm (FV > 150 cm/s). None of the patients with initially normal THRT results developed DIDs, although four patients did exhibit late (secondary) THRT impairment, which was associated with FVs of more than 120 cm/s. CONCLUSION: When the effects of primarily impaired (after surgery) autoregulation are magnified by vasospasm, the risk of DIDs seems to be very high. Vasospasm alone does not seem to cause DIDs. The development of DIDs could therefore be predicted using the THRT for patients after aneurysm clipping. PMID- 11014422 TI - Radiosurgery for childhood intracranial arteriovenous malformations. AB - OBJECTIVE: The optimal management of intracranial arteriovenous malformations (AVMs) in children remains controversial. Children with intracranial AVMs present a special challenge in therapeutic decision-making because of the early recognition of their future life-long risks of hemorrhage if they are treated conservatively. The goals of radiosurgery are to achieve complete AVM obliteration and to preserve neurological function. We present long-term outcomes for a series of children treated using radiosurgery. METHODS: The findings for 53 consecutive children who underwent at least 36 months of imaging follow-up monitoring after radiosurgery were reviewed. The median age at the time of treatment was 12 years (range, 2-17 yr). Thirty-one children (58%) presented after their first intracranial hemorrhaging episodes, two (4%) after their second hemorrhaging episodes, and one (2%) after five hemorrhaging episodes. Nineteen children (36%) presented with unruptured AVMs, and a total of 25 children (47%) exhibited neurological deficits. AVMs were graded as Spetzler-Martin Grade I (2%), Grade II (23%), Grade III (36%), Grade IV (9%), or Grade VI (30%). The median AVM volume was 1.7 ml (range, 0.11-10.2 ml). The median marginal dose was 20 Gy (range, 15-25 Gy). RESULTS: Results were stratified according to AVM volumes (Group 1, < or =3 ml; Group 2, >3 ml to < or =10 ml; Group 3, >10 ml). Twenty-eight patients (80%) in Group 1 and 11 (64.7%) in Group 2 achieved complete obliteration. The only patient in Group 3 did not achieve obliteration. Complications included brainstem edema (n = 1) and transient pulmonary edema (n = 1). Four patients experienced hemorrhaging episodes, 30, 40, 84, and 96 months after radiosurgery. Multivariate logistic regression analysis demonstrated that only volume was significantly correlated with obliteration rates (P = 0.0109). CONCLUSION: Radiosurgery is safe and efficacious for selected children with AVMs. The obliteration rates and the attendant low morbidity rates suggest a primary role for stereotactic radiosurgery for pediatric AVMs. PMID- 11014421 TI - Management outcomes for ruptured and unruptured aneurysms in the elderly. AB - OBJECTIVE: In a patient older than 70 years, the decision to treat an intracranial aneurysm remains difficult whether it is ruptured or unruptured. We sought to review our institutional risks of treatment of such lesions in the context of the risks of rupture and its associated morbidity and mortality in this age group. METHODS: One hundred twenty-nine consecutive patients aged 70 years or older, who were treated at a single institution for an intracranial aneurysm, were retrospectively reviewed. Forty patients were treated for unruptured aneurysms, and 89 patients presented after subarachnoid hemorrhage. Seven additional patients in this age group who had solely intracavernous lesions, as well as one patient with a dolichoectatic fusiform basilar lesion, were excluded. Management outcomes were assessed using a modification of the Glasgow Outcome Scale, and additional physical and functional disability was assessed using the Barthel index and the Reintegration to Normal Living index. RESULTS: Six-month outcomes for the unruptured group were: excellent, 70%; good, 15%; fair, 5%; poor, 7.5%; and death (2.5%). Outcomes for all patients with ruptured lesions (including those not offered aggressive therapy) were: excellent, 34%; good, 9%; fair, 5.6%; poor, 3.4%; and death, 45%. Long-term follow-up was performed by questionnaire to assess physical and functional disability. Although physical disability (Barthel index) was similar among survivors, the Reintegration to Normal Living index, a global assessment of function, was significantly higher in patients with unruptured aneurysms (84.8 versus 70.1; P = 0.05), which highlights the disabling effects of hemorrhage. CONCLUSION: On the basis of an individual treatment center's management risks, annual aneurysmal rupture rates can be estimated that justify treatment in this difficult patient population. Despite recent controversy regarding aneurysmal hemorrhage rates, we think that symptomatic unruptured aneurysms should be treated and good results can be achieved, even in older patients. PMID- 11014424 TI - Hyperthermia in the neurosurgical intensive care unit. AB - OBJECTIVE: In patients with traumatic or ischemic brain injury, hyperthermia is thought to worsen the neurological injury. We studied fever in the neurosurgical intensive care unit (ICU) population using a definition common to surgical practice (rectal temperature >38.5 degrees C). We sought to determine fever incidence, fever duration, and peak temperature and to quantify the use of antipyretic therapy. We also attempted to determine the patient subgroups that are at highest risk for development of fever. METHODS: In a retrospective chart review of a 6-month period, all febrile episodes that occurred in a consecutive series of neurosurgical ICU patients in a university hospital setting were studied. A febrile episode was defined as a rectal temperature of at least 38.5 degrees C; an episode lasted until the temperature fell below this threshold. RESULTS: The 428 patients studied had 946 febrile episodes. Fever occurred in 47% of patients, with a mean of 4.7 febrile episodes in each febrile patient. Fevers occurred in more than 50% of patients who were admitted to the ICU for subarachnoid hemorrhage, a central nervous system infection, seizure control, or hemorrhagic stroke, but they occurred in only 27% of patients admitted for spinal disorders. Fevers occurred in 15% of the patients who stayed in the ICU less than 24 hours, but in 93% of those who remained longer than 14 days. Despite the use of antipyretic therapy for 86% of the febrile episodes, 57% lasted longer than 4 hours and 5% lasted longer than 12 hours. CONCLUSION: Fever is common in critically ill neurosurgical patients, especially those with a prolonged length of stay in the ICU or a cranial disease. If hyperthermia worsens the functional outcome after a primary ischemic or traumatic injury, as has been suggested by several studies of stroke patients, treatment of fever is a clinical issue that requires better management. PMID- 11014423 TI - Surgical treatment of clinically nonsecreting pituitary adenomas in elderly patients. AB - OBJECTIVE: The goal of this study was to evaluate the efficacy and safety of transsphenoidal pituitary surgery for elderly patients, using improved techniques of the past decade. METHODS: We retrospectively analyzed 32 surgically treated cases of clinically nonsecreting pituitary adenomas in patients more than 70 years of age (mean, 73.9+/-3.4 yr). These patients were identified in a review of 982 patients with pituitary adenomas who were treated at University Hospital Eppendorf, Hamburg, between January 1991 and November 1999. RESULTS: The mean preoperative duration of symptoms was 1.9 years (2 wk to 11 yr). The chiasmatic syndrome was present for 27 patients (84.4%). All patients underwent transsphenoidal surgery. Seven patients underwent reoperations. Preoperative assessments of anterior pituitary function revealed growth hormone deficiencies for 21 of 27 patients (77.8%), thyroid insufficiencies for 10 of 30 patients (33.3%), and adrenal insufficiencies for 13 of 29 patients (44.8%). Hypogonadism and hyperprolactinemia were observed for 76.7% and 46.9% of the patients, respectively. All tumors were macroadenomas, ranging from 18 to 50 mm (average, 33.6 mm) in size, including 7 enclosed and 25 invasive adenomas. Complete microscopic tumor resection was achieved in 24 cases, and subtotal removal was performed in 8 cases. There were no severe perioperative complications. In the cases involving hyperprolactinemia, serum prolactin levels were normalized for 8 of 11 patients (72.7%). Normal thyroid function was recovered for 1 of 10 patients (10.0%) with preoperative hypothyroidism. However, growth hormone or adrenal insufficiencies persisted for all patients with preoperative insufficiencies. Visual disturbances were improved for 19 of 23 patients (82.6%). All patients recovered well after surgery, with an average hospital stay of 16.3 days. Histological and immunohistochemical studies demonstrated gonadotroph adenomas in 56.7% of cases, null-cell adenomas in 26.7%, and oncocytomas in 13.3%. CONCLUSION: Surgical treatment of nonsecreting pituitary adenomas causing visual disturbances is standard, even for elderly patients. In this series, transsphenoidal surgery was a safe procedure, with minimal morbidity and excellent tolerance. Age alone is not a contraindication for active treatment, particularly with transsphenoidal surgery. PMID- 11014425 TI - Craniopharyngiomas of the third ventricle: trans-lamina terminalis approach. AB - OBJECTIVE: Craniopharyngiomas usually grow on the cisternal surface of the hypothalamic region; these tumors can also grow from the infundibulum or tuber cinereum on the floor of the third ventricle, developing exclusively into the third ventricle. The aim of the present work was to establish the usefulness of the pterional trans-lamina terminalis approach for the removal of these tumors. METHODS: Eight patients who were surgically treated for craniopharyngiomas located exclusively within the third ventricle were considered. The initial symptoms were acute hydrocephalus in two cases, psychological disturbances in two, amenorrhea in two, headaches in one, and hypopituitarism in one. The diagnoses were established, in all cases except one, with magnetic resonance imaging. In all cases, the tumor completely filled the third ventricle. RESULTS: Total removal of the lesion was achieved in seven cases. One patient underwent partial removal. In the immediate postoperative period, no major complications were observed. Five patients required replacement hormonal therapy. All patients returned to a normal life. Many months after surgery, two patients exhibited psychological disturbances and died, the first because of voluntary withdrawal of replacement therapy (12 mo after surgery) and the second because of a severe imbalance in body fluids and electrolytes, with a subsequent hyperosmolar coma (27 mo after surgery). Only one patient who underwent initial total removal experienced a small recurrence of the lesion (30 mo after surgery); after 3 years, the lesion exhibited unchanged size. CONCLUSION: In our experience, the trans-lamina terminalis approach is a valid choice for the removal of purely intraventricular craniopharyngiomas. These tumors can be removed without significant sequelae related to the surgical approach. The proximity to the hypothalamus requires accurate neuroendocrine and electrolyte control in the postoperative period, in some cases even years after surgery. PMID- 11014426 TI - Three-dimensional reconstructed images after rotational angiography in the evaluation of intracranial aneurysms: surgical correlation. AB - OBJECTIVE: To evaluate the diagnostic accuracy of three-dimensional reconstructed images from rotational digital subtraction angiography in the surgical treatment of intracranial aneurysms. METHODS: Twenty-two patients with 34 intracranial aneurysms underwent biplane angiography (40 degrees per s, 4.5 degrees per image, 8.8 frames per s). Three-dimensional (3-D) reconstructed images were obtained at a separate Advantage 3.1 workstation (General Electric, Milwaukee, WI) after the rotational images were transferred. The available visualization techniques included maximum intensity projection, shaded surface display, and virtual endoluminal view. All images were evaluated in correlation with intrasurgical visual data recorded on digital videotapes. RESULTS: 3-D reconstructed images correlated well with surgical findings. The shape of the aneurysms, their neck size, and their relationships to the parent vessels and other branches were depicted clearly, especially compared with images obtained by two-dimensional conventional digital subtraction angiography and magnetic resonance angiography. CONCLUSION: 3-D digital subtraction angiography enables the surgeon to understand the 3-D structure of lesions and is very useful in planning the surgical treatment of cerebral aneurysms. PMID- 11014427 TI - Computer-assisted fluoroscopic targeting system for pedicle screw insertion. AB - OBJECTIVE: Biplanar fluoroscopic imaging linked to a computer-driven mechanical end-effector is under development as a targeting system for spinal surgery. This technology has the potential to enhance standard intraoperative fluoroscopic information for localization of the pedicle entry point and trajectory, and it may be an effective alternative to the computed tomography-based image-guided system (IGS) in pedicle screw placement. A preclinical study to assess the accuracy and time efficiency of this system versus a conventional IGS was conducted. METHODS: Pedicle screw placement was performed in six cadavers from T1 to S1 levels using the ViewPoint IGS (Picker International, Inc., Cleveland, OH) on one side versus the Fluorotactic guidance system (Z-Kat, Inc., Miami, FL) on the other side. Of 216 possible pedicles, 208 were instrumented; 8 pedicle diameters were too small or were not adequately imaged. Postinsertion, each pedicle was assessed for the presence and location of cortical perforation using computed tomographic scanning and direct visualization. RESULTS: The number of successful screw placements was 89 (87.3%) of 102 for IGS and 87 (82.1 %) of 106 for the Fluorotactic guidance system, respectively. The mean time to register and operate on one level using the Fluorotactic guidance system was 14:34 minutes (minutes:seconds), compared with 6:50 minutes using the IGS. The average fluoroscope time was 4.6 seconds per pedicle. CONCLUSION: Our data indicate that this first-generation fluoroscopy-based targeting system can significantly assist the surgeon in pedicle screw placement. The overall accuracy is comparable to an IGS, especially in the region of T9-L5. A second-generation system with a faster end-effector and user-friendly interface should significantly reduce the operating and fluoroscope time. PMID- 11014428 TI - Correlation of neurosurgical subspecialization with outcomes in children with malignant brain tumors. AB - OBJECTIVE: This study was performed to evaluate the association between the type of neurosurgeon (general or pediatric) and either the extent of tumor removal or the frequency of complications in children undergoing malignant brain tumor resections. METHODS: Data were analyzed from three recent Children's Cancer Group studies: two on medulloblastomas/primitive neuroectodermal tumors and one on malignant gliomas. Neurosurgeons were classified as general neurosurgeons, as designated pediatric neurosurgeons in their institutions, or as members of the American Society of Pediatric Neurosurgeons (ASPN), which requires pediatric neurosurgical experience and practice standards. RESULTS: Data forms from 732 children were analyzed; 485 were from children with medulloblastomas/primitive neuroectodermal tumors, and 247 were from children with malignant gliomas. Operations were performed by 269 neurosurgeons, including 213 general neurosurgeons, 29 designated pediatric neurosurgeons, and 27 ASPN members. The mean number of operations per surgeon was 1.8, 4.9, and 7.6 for general neurosurgeons, designated pediatric neurosurgeons, and ASPN members, respectively. There was a significant relationship between the extent of tumor resection or the amount of residual tumor and the type of neurosurgeon. Designated pediatric neurosurgeons and ASPN members were more likely to remove more than 90% of the tumor and to leave less than 1.5 cc of residual tumor than were general neurosurgeons (P<0.05). In these studies, the probability of extensive tumor removal correlated with the number of operations the neurosurgeon performed (P<0.01). Neurological complications occurred in the following proportion of cases: general neurosurgeons, 23%; designated pediatric neurosurgeons, 32%; and ASPN members, 18%. CONCLUSION: Pediatric neurosurgeons are more likely than general neurosurgeons to extensively remove malignant pediatric brain tumors. In these tumors, extent of removal has been demonstrated to influence survival. PMID- 11014429 TI - Molecular insight into medulloblastoma and central nervous system primitive neuroectodermal tumor biology from hereditary syndromes: a review. AB - Through the study of uncommon familial syndromes, physicians and scientists have been able to illuminate the underlying mechanisms of some of the more common sporadic diseases; this is illustrated best by studies of familial retinoblastoma. A number of rare familial syndromes have been described in which affected individuals are at increased risk of developing medulloblastoma and/or supratentorial primitive neuroectodermal tumors. The descriptions of many of these syndromes are based on patients observed by clinicians in their clinical practice. Determination of the underlying genetic defects in these patients with uncommon syndromes has led to identification of a number of genes subsequently found to be mutated in sporadic medulloblastomas (tumor suppressor genes). Associated genes in the same signaling pathways have also been found to be abnormal in sporadic medulloblastoma. Identification of patients with these rare syndromes is important, as they are often at increased risk for additional neoplasms, as are family members and future children. We review the published literature describing hereditary syndromes that have been associated with an increased incidence of medulloblastoma and/or central nervous system primitive neuroectodermal tumor. Review of the underlying molecular abnormalities in comparison to changes found in sporadic neoplasms suggests pathways important for tumorigenesis. PMID- 11014430 TI - Liliequist's membrane is a fold of the arachnoid mater: study using sheet plastination and scanning electron microscopy. AB - OBJECTIVE: The subarachnoid space consists of a number of distinct subarachnoid cisterns. They are separated from each other by trabecular walls, one of which is Liliequist's membrane. The aim of this study was to investigate the anatomic characteristics of Liliequist's membrane. METHODS: The study used a combined approach, consisting of the modified E12 sheet plastination method for 3 adult cadavers and gross anatomic dissection for 35 cadavers, 2 of which were further examined using scanning electron microscopy. RESULTS: The results from this study indicate that 1) Liliequist's membrane is an avascular fold of the arachnoid mater that lacks openings and spreads out laterally, being in direct continuity with the arachnoid mater covering the tentorium; 2) the carotid-chiasmatic walls, which separate the chiasmatic cistern and carotid cisterns and had been considered to be parts of Liliequist's membrane, are vascular and incomplete trabecular walls and should not be considered parts of Liliequist's membrane; and, 3) as a fold of the arachnoid mater, Liliequist's membrane is not directly attached to the temporal lobes and oculomotor nerves. CONCLUSION: Liliequist's membrane is a double-layer fold of the arachnoid mater and has anatomic characteristics different from those of arachnoid trabecular walls. PMID- 11014431 TI - Abnormal pattern of Tie-2 and vascular endothelial growth factor receptor expression in human cerebral arteriovenous malformations. AB - OBJECTIVE: Human cerebral arteriovenous malformations (AVMs) are speculated to result from abnormal angiogenesis. Vascular endothelial growth factor receptors (VEGF-Rs) and Tie-2 play critical roles in vasculogenesis and angiogenesis. We hypothesized that the abnormal vascular phenotype of AVMs may be associated with abnormal expression of VEGF-Rs and Tie-2. METHODS: We measured the expression of Tie-2, VEGF-R1, and VEGF-R2 in AVMs and normal brain tissue, using immunoblotting. To assess active vascular remodeling, we also measured endothelial nitric oxide synthase expression. CD31 expression was used to control for endothelial cell mass for Tie-2, VEGF-Rs, and endothelial nitric oxide synthase. Immunoblotting data were presented as relative expression, using normal brain tissue values as 100%. RESULTS: CD31 was expressed to similar degrees in AVMs and normal brain tissue (99+/-29% versus 100+/-20%, mean +/- standard error, P = 0.98). Tie-2 expression was markedly decreased in all AVMs, compared with normal brain tissue (16+/-9% versus 100+/-37%, P = 0.04). VEGF-R1 expression was decreased in four of five AVMs, but the difference between the mean values was not significant (35+/-8% versus 100+/-42%, P = 0.14). VEGF-R2 expression was decreased in all AVMs, compared with normal brain tissue (28+/-6% versus 100+/ 29%, P = 0.03). There was no difference in endothelial nitric oxide synthase expression between AVMs and normal brain tissue (106+/-42% versus 100+/-25%, P = 0.91). CONCLUSION: AVM vessels exhibited abnormal expression of Tie-2 and VEGF Rs, both of which may contribute to the pathogenesis of AVMs. PMID- 11014432 TI - Vascular endothelial growth factor-driven glioma growth and vascularization in an orthotopic rat model monitored by magnetic resonance imaging. AB - OBJECTIVE: The goal of the present study was to develop an orthotopic in vivo model for the investigation of vascular endothelial growth factor (VEGF) dependent glioma growth and vascularization. METHODS: C6 glioma cells were infected with viruses encoding sense or antisense VEGF. Expression of the transgene was controlled by Northern blot analysis, Western blot analysis, and immunohistochemistry. Spheroids generated from both clones as well as from wild type and mock-transfected cells were implanted in the brains of Sprague-Dawley rats. Growth and vascularization were assessed using magnetic resonance imaging after 7 and 11 days. Histology was studied using hematoxylin and eosin staining, immunohistochemistry with anti-von Willebrand staining, anti-VEGF, anti-CD8, and assessment of vessel density. RESULTS: Cell proliferation, migration, and invasion in vitro were very similar in all cell clones. Sense gliomas demonstrated by far the fastest growth in vivo, with intense contrast enhancement meeting criteria for highly malignant tumors. Histological examination revealed masses of von Willebrand- and VEGF-positive tumor vessels with a high vessel density. Antisense gliomas depicted the radiological features of low-grade gliomas, with slow growth and poor vascularization, although they were highly infiltrative. Wild-type and mock-transfected gliomas demonstrated similar growth and vascularization patterns intermediate between sense and antisense gliomas. Any influence of the allogeneic response of the hosts on different tumor sizes could be excluded. CONCLUSION: Our model elucidates glioma growth and vascularization as strongly VEGF dependent, which is consistent with human gliomas. Thus, this model is suitable for testing antiangiogenic strategies to interfere with the VEGF/VEGF receptor system, as well as for exploring VEGF independent mechanisms using the antisense-transfected clone. PMID- 11014434 TI - Preconquest Peruvian neurosurgeons: a study of Inca and pre-Columbian trephination and the art of medicine in ancient Peru. AB - Trephination and craniotomy performed by abrasion, scraping, crosscut sawing, and drilling are the oldest known surgical techniques used by primitive peoples. As a result of archaeological findings, the human skull is the most frequently studied part of the excavated body, leading to the creation of a new aspect of anthropology known as "cultural osteology." Found in ancient tombs, the human remains, mummies, skeletons, and their belongings, including war instruments, pottery, clothing, jewels, and surgical instruments, constitute the richest source of insight into the lives and pragmatic activities of ancient cultures. This study summarizes thousands of years of pre-Columbian history and medical evolution, specifically in the early and primitive practice of trephination, as precursors of neurosurgery. Comparative osteology studies have demonstrated that using primitive stone or metal instruments, the sirkaks (Inca surgeons) achieved an average survival rate of 50 to 70% of their craniectomy patients, with little incidence of infection or other complications. Despite their rudimentary knowledge of disease and pathology, a considerable knowledge of anatomy and natural medicine provided them with hemostatic agents, antiseptics, and other medical drugs, such as quinine for fever and malaria, as well as gold, silver, and other products to perform cranioplasties. Living in a world of continuous hand-to-hand combat, they also developed aggressive and defensive weapons that necessitated refinement of surgical techniques to save soldiers from battle wounds to their poorly protected crania. PMID- 11014433 TI - Combined antitumor effects of an adenoviral cytosine deaminase/thymidine kinase fusion gene in rat C6 glioma. AB - OBJECTIVE: In this study, we investigated the feasibility of a double-suicide gene/prodrug therapy, involving direct introduction of the herpes simplex virus Type 1 thymidine kinase (TK) gene and the Escherichia coli cytosine deaminase (CD) gene, via a recombinant adenoviral vector, and ganciclovir (GCV) and/or 5 fluorocytosine (5-FC) treatment, in a rat C6 glioma model. METHODS: Efficient gene transfer and transduction of C6 glioma cells via a recombinant adenovirus were evaluated by infecting cells with adenovirus bearing the beta-galactosidase gene and then staining cells with X-5-bromo-4-chloro-3-indolyl-13-D-galactoside. CD/TK expression in cells infected with adenovirus bearing the CD/TK gene (ad CD/TK) was examined by immunoblotting analysis. For in vitro cytotoxicity experiments, the cells were infected with ad-CD/TK or ad-deltaE1 (as a control). After the addition of a variety of concentrations of GCV and 5-FC, either separately or in combination, cell viability was determined by staining the cells with crystal violet solution 6 days after infection. For in vivo antitumor experiments, 1x10(5) cells were stereotactically injected into the right caudate putamen of female Wistar rat brains. At 3 days after implantation, 1x10(8) plaque forming units of ad-CD/TK or ad-deltaE1 (as a control) were stereotactically injected into the tumors and GCV (25 mg/kg) and 5-FC (250 mg/kg), alone or in combination, were intraperitoneally administered. Animals were then killed, and tumor volumes were measured by determining the tumor area in every fifth section, using a light microscope. RESULTS: C6 glioma cells were efficiently transduced with recombinant adenoviral vector at multiplicities of infection of 200 or more. In vitro cytotoxicity of GCV and/or 5-FC, either alone or in combination, was exclusively observed in the cells transduced with ad-CD/TK. Obvious cytotoxicity (>50% inhibition) was observed in the presence of 5-FC at concentrations greater than 30 microg/ml or GCV at concentrations greater than 0.3 microg/ml at a multiplicity of infection of 100. Additionally, cytotoxicity in the presence of both GCV and 5-FC was greater than that after single-prodrug treatments, indicating additive effects of the prodrug treatments. In in vivo experiments, the tumor volumes of the rats treated with GCV or 5-FC alone after ad-CD/TK injection (59.1+/-4.6 and 57.4+/-7.1 mm3, respectively) were significantly smaller than that of the control rats (157+/-8.9 mm3, P<0.05). Furthermore, the tumor volume of the rats treated with GCV and 5-FC in combination was 14.7+/-1.8 mm3. CONCLUSION: These results demonstrated the efficient transduction of C6 glioma cells with a recombinant adenovirus and the additive effects of CD/TK fusion gene/GCV/5-FC treatment, compared with single-gene therapy with the TK or CD gene. Therefore, our data suggest that the direct administration of a double suicide gene/prodrug therapy has great potential in the treatment of brain tumors. PMID- 11014435 TI - Color illustrations and neurosurgical techniques of Serefeddin Sabuncuoglu in the 15th century. AB - Serefeddin Sabuncuoglu (AD 1385-1468?) is the author of Cerrahiyyetu'l-Haniyye (Imperial Surgery), which was written in Turkish in 1465. It was the first illustrated textbook of surgery in the Turkish medical literature, containing color illustrations of surgical procedures, incisions, and instruments. Sabuncuoglu, a pioneer of surgery, developed numerous original techniques in a variety of surgical specialties. He described surgical management of spinal trauma, epilepsy, migraine, facial palsy, hemiplegia, low back pain, cranial fracture, and hydrocephalus. The aim of this study is to describe his contributions to neurosurgery. PMID- 11014436 TI - Sclerosing epithelioid fibrosarcomas involving the neuraxis: report of three cases. AB - OBJECTIVE AND IMPORTANCE: Sclerosing epithelioid fibrosarcoma (SEF) is a rare mesenchymal neoplasm composed of rounded, vimentin-immunoreactive tumor cells disposed in nests and cords within a hyalinized collagenous matrix. Most examples arise in the deep skeletal muscles of adults. The cases recorded to date have been characterized by protracted clinical evolutions with a tendency for stubborn local recurrence, followed by late metastasis. Accordingly, SEF has been regarded as a low-grade sarcoma. A single instance of brain and vertebral metastasis has been described. We report three examples of SEF distinguished by primary involvement of the neuraxis at initial presentation. CLINICAL PRESENTATION: Two tumors had intracranial, calvarial and extracalvarial, soft-tissue components, whereas the third tumor manifested as a paraspinal mass with extension into the T12-L1 neural foramen and invasion of the T12 nerve root. INTERVENTION: All three affected patients experienced local recurrence and distant metastasis after resection of the primary site. These complications appeared early in the disease course in two cases. In no case was there a response to adjuvant chemotherapy or radiotherapy. CONCLUSION: Our experience indicates that SEFs arising along the neuraxis may demonstrate unexpectedly aggressive clinical behavior, compared with those arising in the more typical location of deep skeletal muscles. PMID- 11014437 TI - Intravascular papillary endothelial hyperplasia arising within a posteroinferior cerebellar artery aneurysm: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Intravascular papillary endothelial hyperplasia (IPEH) is an atypical proliferation of endothelium that results in abnormal organization for thrombus formation. Intracranial IPEH is a rare entity and has not been reported to arise from within an intracranial aneurysm. Furthermore, the elapsed time during which acquired intracranial IPEH develops has not been previously documented. CLINICAL PRESENTATION: In this case report, a patient with facial and neck pain was noted to have an enhancing mass lesion lateral to the medulla in magnetic resonance imaging scans. Angiography revealed a vascular mass adjacent to the posteroinferior cerebellar artery. Normal magnetic resonance imaging and magnetic resonance angiographic findings had been obtained for the patient 29 months earlier. INTERVENTION: During surgery, a thrombosed, 2.5-cm, posteroinferior cerebellar artery aneurysm was resected and noted to contain florid IPEH. There has been no evidence of recurrence in 1 year of follow-up monitoring. A literature search revealed 13 cases of intracranial IPEH, in which recurrence was observed for incompletely resected lesions. CONCLUSION: IPEH can develop in a relatively short time, can present as a hypervascular mass lesion or within an intracranial aneurysm, and should be completely resected to prevent recurrence. PMID- 11014438 TI - Bilateral cerebellopontine angle arachnoid cysts: case report. AB - OBJECTIVE AND IMPORTANCE: A rare case of bilateral cerebellopontine angle (CPA) arachnoid cysts (ACs), accompanied by cerebellar tonsillar displacement toward the foramen magnum, is presented. CLINICAL PRESENTATION: A 45-year-old woman presented with progressive dysphagia, vertigo, and truncal ataxia. Magnetic resonance imaging revealed bilateral CPA ACs and cerebellar tonsillar displacement. INTERVENTION: The right CPA AC was excised via a suboccipital approach. Decompression of the foramen magnum and duraplasty were also performed. CONCLUSION: The case reported here is the first case of bilateral CPA ACs. Decompression of the foramen magnum and excision of the cyst resulted in complete relief of symptoms. PMID- 11014439 TI - Tuberculosis of the axis in a patient with systemic sarcoidosis: technique of posterior open biopsy of the dens: case report. AB - OBJECTIVE AND IMPORTANCE: This case report illustrates the importance of obtaining tissue from a destructive lesion of the dens in a patient with systemic sarcoidosis. Although sarcoidosis can involve the axial skeleton, tissue obtained at the time of C1-C2 fusion demonstrated unsuspected pathological features, which dramatically altered the subsequent medical treatment. The technique of open posterior biopsy of the dens is illustrated, and the advantages of the approach are discussed. CLINICAL PRESENTATION: A 40-year-old woman with systemic sarcoidosis developed neck pain and atlantoaxial instability. Imaging revealed multiple thoracic and cervical vertebral abnormalities, including a destructive enhancing lesion involving the base of the dens. INTERVENTION: At the time of posterior C1-C2 fusion, we elected to perform an open biopsy of the base of the dens. A 16-gauge biopsy needle was introduced along the medial portion of the left C2 pars, aiming medially toward the base of the odontoid process. This procedure was performed under direct observation, with fluoroscopic guidance. The biopsy specimen contained caseating granulomas, and cultures were positive for Mycobacterium tuberculosis. CONCLUSION: The unusual presentation, the technique, and the importance of obtaining tissue to confirm the diagnosis of tuberculous involvement of the dens are emphasized. The relationship between sarcoidosis and tuberculosis reported in the literature is reviewed. In the current case, cell wall-positive tuberculous bacteria were cultured, confirming the presence of two separate diseases in the same patient. PMID- 11014440 TI - Candida albicans cerebral granulomas associated with a nonfunctional cerebrospinal fluid shunt: case report. AB - OBJECTIVE AND IMPORTANCE: We report an unusual case of basal ganglia granulomas caused by Candida albicans that surrounded the proximal segment of a nonfunctional cerebrospinal fluid shunt in a previously healthy patient. CLINICAL PRESENTATION: A 22-year-old woman had undergone ventriculoatrial cerebrospinal fluid shunt placement for posttraumatic hydrocephalus 3 years previously. One year later, a shunt revision was followed by wound dehiscence with local infection at the neck level. She received oral administration of antibiotics for 3 months until the wound closed. Twelve weeks before admission, the patient experienced pulmonary emboli. She received anticoagulants, and the distal segment of the shunt was removed. Five weeks after shunt removal, she presented with headache and left-sided hemiplegia caused by right basal ganglia inflammatory masses. INTERVENTION: A stereotactic brain biopsy was performed, and the shunt remnants were removed. Microscopically, the lesions were acutely and chronically inflamed. C. albicans grew in tissue and in shunt hardware cultures. The patient was treated with 1.1 g of intravenously administered amphotericin B and orally administered ketoconazole; she recovered completely. CONCLUSION: C. albicans brain granulomas occur rarely in immunocompetent patients. Despite the large size of the lesions and severe brain edema, the absence of an underlying disease contributed to complete resolution after shunt removal and antifungal therapy. PMID- 11014441 TI - Chronic hydrocephalus presenting with bilateral ptosis after minor head injury: case report. AB - OBJECTIVE AND IMPORTANCE: Some patients with hydrocephalus may exhibit various signs of oculomotor dysfunction. However, ptosis has not previously been described in chronic hydrocephalus patients. CLINICAL PRESENTATION: We report a 50-year-old woman who was diagnosed with chronic hydrocephalus based on an evaluation for bilateral ptosis after a minor head injury. She exhibited bilateral ptosis and upward gaze paralysis, but other oculomotor functions were normal. Neuroimages revealed chronic hydrocephalus with no traumatic abnormalities. INTERVENTION: The eyelid dysfunction resolved after placement of a right ventriculoperitoneal shunt with a programmable pressure valve. CONCLUSION: The resolution of eyelid dysfunction by cerebrospinal fluid diversion suggests that chronic hydrocephalus was involved in the development of ptosis after the minor head injury. A mild but sudden cerebrospinal fluid pressure change at the time of minor head injury might induce functional impairment at the level of vulnerable periaqueductal structures, which barely withstood the longstanding ventriculomegaly, resulting in the clinical features observed in our patient. PMID- 11014442 TI - Carotid stenting and "extarterectomy" in the management of head and neck cancer involving the internal carotid artery: technical case report. AB - OBJECTIVE AND IMPORTANCE: Head and neck cancer that invades the internal carotid artery (ICA) represents a significant management challenge. We describe a novel technique that allows for aggressive tumor removal without disrupting blood flow through the affected ICA. CLINICAL PRESENTATION: A 62-year-old man was referred to our institution for management of a neck malignancy involving the ICA. Cerebral angiography suggested that there was good collateral flow from the opposite hemisphere, but the patient reported visual loss in the ipsilateral eye during balloon test occlusion of the ICA. INTERVENTION: A self-expanding stent was deployed in the ICA; it spanned the entire length of the artery involved by tumor. One month later, the patient underwent tumor resection. During surgery, a long ICA arteriotomy was performed directly down to the mesh of the stent. A neoendothelium had formed within the stent, which prevented arterial bleeding. The carotid wall was dissected from the stent without difficulty and removed en bloc with the surrounding tumor. The exposed stent was wrapped circumferentially with a synthetic patch material. The patient tolerated the procedure well, and postoperative angiography demonstrated normal filling of the ICA. CONCLUSION: We describe a novel approach to a patient with head and neck cancer involving the cervical ICA. Preliminary stenting, which allows time for endothelialization before surgery, may permit aggressive tumor resection without interrupting flow through the ICA. This technique obviates the need for complicated carotid reconstruction procedures and avoids the risk of delayed ischemia from carotid sacrifice. PMID- 11014443 TI - Modified malis bayonet forceps aids application of the cyberonics vagus nerve stimulator electrode: technical note. AB - OBJECTIVE: To notify neurosurgeons about a modified bayonet forceps that aids application of the vagus nerve stimulating electrode. METHODS: The manufacturer (Codman & Shurtleff, Inc., Raynham, MA) extended the tips of an upward-angled Malis bayonet forceps from 2 mm to 6 mm. RESULTS: The modified bayonet tips, when placed under the vagus nerve, extend well beyond the edge of the usual vagus nerve to easily accept the electrode lead. CONCLUSION: The modified bayonet forceps and depicted wrapping sequence shorten electrode wrapping time. PMID- 11014444 TI - Limitations of the C6/Wistar rat intracerebral glioma model: implications for evaluating immunotherapy. AB - OBJECTIVE: Intracranial rat glioma models are a useful method for evaluating the efficacy and toxicity of novel therapies for malignant glioma. The C6/Wistar model has been used extensively as a reproducible in vivo model for studying primary brain tumors including anti-glioma immune responses. The objective of the present study is to provide in vivo evidence that the C6 rat glioma model is allogeneic within Wistar rats and is therefore inappropriate for evaluating immune responses. METHODS: Growth patterns and immune responses of C6 cells implanted into the brain and flank of Wistar rats were analyzed and compared to an immunogenic syngeneic model (9L/Fischer). RESULTS: Wistar rats with C6 tumors developed a potent humoral and cellular immune response to the tumor. Wistar rats given simultaneous flank and intracerebral tumors had a survival rate of 100% compared to an 11% survival rate in control animals receiving only intracranial C6 cells. CONCLUSION: The C6 rat glioma induces a vigorous immune reaction that may mimic a specific anti-tumor response in Wistar rats. Efficacy of immunotherapy within this model must be cautiously interpreted. PMID- 11014445 TI - Stable isotope incorporation triples the upper mass limit for determination of elemental composition by accurate mass measurement. AB - By comparing electrospray ionization Fourier-transform ion cyclotron resonance (FT-ICR) mass spectra and collision-induced dissociation (CID) FT-ICR mass spectra of a phospholipid (851 Da) extracted from natural abundance and 99% 13C bacterial growth media, we are able to reduce its number of possible elemental compositions (based on +/-10 ppm externally calibrated mass accuracy and biologically relevant compositional constraints) from 394 to 1. The basic idea is simply that the mass of a molecule containing N carbon atoms increases by N Da when 12C is replaced by 13C. Once the number of carbons is known, the number of possible combinations of other atoms in the molecule is greatly reduced. We demonstrate the method for a stored-waveform inverse Fourier transform-isolated phospholipid from an extract of membrane lipids from Rhodococcus rhodochrous hydrocarbon-degrading bacteria grown on either natural abundance or 99% 13C enriched mixtures of n-hexadecane and n-octadecane. We project that this method raises the upper mass limit for unique determination of elemental composition from accurate mass measurement by a factor of at least 3, thereby extending "chemical formula" determination to identification and sequencing of larger synthetic and bio-polymers: phospholipids, oligopeptides of more than three to four amino acids, DNA or RNA of more than two nucleotides, oligosaccharides of more than three sugars, etc. The method can also be extended to determination of the number of other atoms for which heavy isotopes are available (e.g., 15N, 34S, 18O, etc.). PMID- 11014446 TI - Mass-correlated pulsed extraction: theoretical analysis and implementation with a linear matrix-assisted laser desorption/ionization time of flight mass spectrometer. AB - The pulsed extraction (PE) of ions produced by matrix-assisted laser desorption/ionization in time-of-flight mass spectrometers greatly improves mass resolution but, unfortunately, this method is mass dependent. Here we report an approach to expand the capabilities of the PE method so as to provide uniform focusing conditions over a wide mass range. Along with an extraction pulse, an additional pulse is applied to correct the mass dependency of the standard PE method. We describe the algorithm for derivation of this correction pulse waveform, where the first-order focusing conditions are valid all along the mass region of interest. Experimental verification of this method for correction of ion velocities demonstrated better mass resolution than standard PE over a wide mass range. PMID- 11014447 TI - Matrix-assisted laser desorption/ionization mass spectrometry methods for oligodeoxynucleotides: improvements in matrix, detection limits, quantification, and sequencing. AB - A comatrix of anthranilic acid and nicotinic acid is optimum for the matrix assisted laser desorption/ionization time of flight determination of oligodeoxynucleotides that are comprised of up to 21 nucleotides. A detection limit of approximately 200 amol was obtained for an oligonucleotide 21mer. The comatrix system is also suitable for quantification of oligodeoxynucleotides provided an internal standard having one more or less nucleotide than the number in the analyte is used. Furthermore, the matrix, when used in combination with the ladder method of sequencing, allows the complete sequence of tens of picomoles of model oligodeoxynucleotides to be determined. PMID- 11014448 TI - Selected ion flow tube mass spectrometry analyses of stable isotopes in water: isotopic composition of H3O+ and H3O+ (H2O)3 ions in exchange reactions with water vapor AB - A new method has been developed for the determination of the isotope abundance ratios of deuterium, D, and oxygen-18, 18O, in water vapor (and water) using selected ion flow tube mass spectrometry (SIFT-MS). H3O+ ions are injected into the helium carrier gas where they associate with the H2O and HDO molecules in a sample of water introduced into the carrier gas. The D and 18O contents of the product cluster ions H8DO4+ and H9(18)OO3+ at m/e = 74 and 75, respectively, are determined by reference to the majority cluster ion H9O4+ at m/e = 73. Allowance is made for the contribution of the H8(17)OO3+ ions to the m/z = 74 ions. Absolute isotopic ratios are measured within seconds without the need for precalibration of the SIFT-MS instrument, currently to an accuracy of better than 2%. PMID- 11014450 TI - Evaluation of alkali metal binding selectivities of caged aza-crown ether ligands by microelectrospray ionization/quadrupole ion trap mass spectrometry AB - Microelectrospray ionization mass spectrometry (MESI-MS) is used to evaluate alkali metal binding selectivities of a variety of macrocyclic compounds. Well studied crown ethers are used to validate the MESI-MS method. A quantitative correlation between MESI mass spectral ion intensities and solution equilibrium distributions of complexes is obtained for the mixtures containing a single host and different alkali metal guest ions. The MESI-MS method is successfully applied for the determination of the alkali metal binding selectivities of a series of cage-functionalized aza-crown ethers and relevant model compounds in methanol and chloroform solutions. The binding selectivities parallel previous results obtained using conventional spectrophotometric extraction methods. Structural differences in the host compounds, such as the presence of a cage functionality, binding cavity size, and overall flexibility, cause significant changes in the binding selectivities. PMID- 11014449 TI - A dual electrospray ionization source combined with hexapole accumulation to achieve high mass accuracy of biopolymers in Fourier transform ion cyclotron resonance mass spectrometry. AB - A dual electrospray ionization (ESI) source employed with hexapole accumulation and gated trapping provides a novel method of using an internal standard to achieve high mass accuracies in Fourier transform ion cyclotron resonance mass spectrometry. Two ESI emitters are sequentially positioned in front of the heated metal capillary inlet by a solenoid fitted to an XYZ micromanipulator; one emitter contains the analyte(s) of interest and the other an internal standard. A 5 V transistor-transistor logic pulse from the data station controls the solenoid by means of a solid-state relay so that matching of spectral peak intensities (i.e., analyte and internal standard intensities) can be accomplished by adjusting the hexapole accumulation time for each species. Polythymidine, d(pT)18, was used as the internal standard for all studies reported here. The absolute average error for an internally calibrated 15-mer oligonucleotide (theoretical monoisotopic mass = 4548.769 Da) was -1.1 ppm (external calibration: 41 ppm) with a standard deviation of +/-3.0 ppm (external calibration: +/-24 ppm) for a total of 25 spectra obtained at various hexapole accumulation time ratios. Linear least squares regression analysis was carried out and revealed a linear dependence of the magnitudes of the peak height ratios (analyte/internal standard) vs. hexapole accumulation time ratios (analyte/internal standard) which is described by the following equation: y = 0.45 x - 0.02. The fitted line had a %RSD of the slope of 28% with an R2 of 0.93. The applicability of this methodology was extended to a polymerase chain reaction product with a theoretical average molecular mass of 50,849.20 Da. With the internal standard, d(pT)18, an absolute average error of -8.9 ppm (external calibration: 44 ppm) based on five measurements was achieved with a standard deviation of 11 ppm (external calibration: +/-36 ppm), thus illustrating this method's use for characterizing large biomolecules such as those encountered in genomics and proteomics related research. PMID- 11014451 TI - Charge-remote and charge-driven fragmentation processes in diacyl glycerophosphoethanolamine upon low-energy collisional activation: a mechanistic proposal. AB - A mechanistic study of diacyl glycerophosphoethanolamine fragmentation under low energy collision-activated dissociation with electrospray ionization tandem mass spectrometry is reported. The fragmentation pathways leading to the formation of carboxylate anions (RxCO2-) (x = 1, 2) and the formation of the ions representing neutral loss of ketene ([M - H - Rx'CH=C=O]-) are charge-driven processes, which are governed by the gas-phase basicity and the steric configuration of the molecules. The fragmentation pathway for the formation of the [M - H - RxCO2H]- ions, reflecting neutral loss of fatty acid, is a charge-remote process, which involves the participation of the hydrogens at C-1 and C-2 of the glycerol, resulting in [M - H - R2CO2H]- > [M - H - R1CO2H]-. The preferential formations of R2CO2- > R1CO2-, and of [M - H - R2'CH=C=O]- > [M - H - R1'CH=C=O]- are attributed to the findings that charge-driven processes are sterically more favorable at sn-2. The observation of the abundance of [M - H - Rx'CH=C=O]- > [M H - RxCO2H]- is attributed to the fact that the [M - H]- ions of GPE are basic precursor ions, which undergo preferential loss of ketene than loss of acid. The major pathway for the formation of RxCO2- ions arises from the nucleophilic attack of the anionic charge site of the phosphate on the C-1 or C-2 of the glycerol to render a charge transfer. The sterically more favorable attack on the C-2 than C-2 of the glycerol results in the abundance of R2CO2- > R1CO2-. These features of tandem spectra readily identify and locate the fatty acid substituents of GPE in the glycerol backbone. PMID- 11014452 TI - Electrospray mass spectrometry and fragmentation of N-linked carbohydrates derivatized at the reducing terminus. AB - Derivatives were prepared from N-linked glycans by reductive amination from 2 aminobenzamide, 2-aminopyridine, 3-aminoquinoline, 2-aminoacridone, 4-amino-N-(2 diethylaminoethyl)benzamide, and the methyl, ethyl, and butyl esters of 4 aminobenzoic acid. Their electrospray and collision-induced dissociation (CID) fragmentation spectra were examined with a Q-TOF mass spectrometer. The strongest signals were obtained from the [M + Na]+ ions for all derivatives except sugars derivatized with 4-amino-N-(2-diethylaminoethyl)benzamide which gave very strong doubly charged [M + H + Na]2+ ions. The strongest [M + Na]+ ion signals were obtained from the butyl ester of 4-aminobenzoic acid and the weakest from 2 aminopyridine. The most informative spectra were recorded from the [M + Li]+ or [M + Na]+ ions. These spectra were dominated by ions produced by sequence revealing glycosidic cleavages and "internal" fragments. Linkage-revealing cross ring cleavage ions were reasonably abundant, particularly from high-mannose glycans. Although the nature of the derivative was found to have little effect upon the fragmentation pattern, 3-aminoquinoline derivatives gave marginally more abundant cross-ring fragments than the other derivatives. [M + H]+ ions formed only glycosidic fragments with few, if any, cross-ring cleavage ions. Doubly charged molecular ions gave less informative spectra; singly charged fragments were weak, and molecular ions containing hydrogen ([M + 2H]2+ and [M + H + Na]2+) fragmented as the [M + H]+ singly charged ions with no significant cross-ring cleavages. PMID- 11014453 TI - Detection and quantification of the sulfated disaccharides in chondroitin sulfate by electrospray tandem mass spectrometry. AB - A new method of identifying and quantifying the disaccharide building blocks of glycosaminoglycans is introduced. The polysaccharides are subjected to an enzymatic digestion that releases the sulfated disaccharides. The disaccharides are then identified using a combination of electrospray ionization mass spectrometry and tandem mass spectrometry. Quantification of the isomeric disaccharides is also achieved by tandem mass spectrometry, using a recently developed methodology which quantifies mixtures of isomers without the use of chromatography or prior separation. Using mass spectrometry to characterize the components of glycosaminoglycans significantly reduces both sample consumption and analysis time of traditional methods. PMID- 11014454 TI - Review of the 48th ASMS conference on mass spectrometry and allied topics held in Long Beach, California June 11-15, 2000 PMID- 11014455 TI - Taxonomy of pain. AB - Research on the pathophysiology of chronic pain has begun to challenge the traditional diagnostic and treatment paradigms for the patient with neuropathic pain. The heterogeneous nature of neuropathic pain indicates that more than one anatomic lesion is most likely responsible for the clinical presentation of a particular syndrome. Numerous pharmacologic agents that have shown improved efficacy in the treatment of neuropathic pain have been developed over the past decade. For the practicing clinician, an important concern is whether the current paradigm for classification of neuropathic pain syndromes is comprehensive enough to address this rapidly expanding body of knowledge. PMID- 11014456 TI - Chronic neuropathic pain: mechanisms and treatment. AB - There have been considerable advances in our understanding of the pathophysiology of neuropathic pain. There is still a lack of consensus about the optimal therapeutic strategy of such conditions, however. Drugs are generally selected on the basis of their established efficacy in randomized controlled studies in etiologically based groups of patients. These studies have been important in confirming the efficacy of antidepressants, antiepileptics, local anesthetics and derivatives, capsaicin, opioids, and N-methyl-D-aspartate antagonists in neuropathic pain, specifically painful diabetic neuropathy and postherpetic neuralgia. More specific therapeutic strategies based on precise quantified assessment of the various components of neuropathic pain are now increasingly used and may provide insight regarding the effects of treatments of particular symptoms (e.g., allodynia, hyperalgesia). In some cases, such assessment may also help to analyze the mechanisms involved in pain, thus allowing selection of treatment on a more rational basis. A mechanism-based approach seems promising for clinical research studies, although its application in current management remains challenging. PMID- 11014457 TI - Neural mechanisms of cutaneous nociceptive pain. AB - Acute mechanical, thermal, and chemically induced pains in the skin are signalled by a set of specific nociceptive afferents, which encode the magnitude of the perceived pain by their discharge intensity. After tissue injury or inflammation, a number of changes in the properties of the primary afferent occur parallel to profound changes in the central nervous system. Primary hyperalgesia (within the area of tissue injury) is best explained by changes of the properties of primary nociceptive afferents, whereas secondary hyperalgesia (increased pain sensitivity outside the area of tissue injury) critically requires functional changes in the central nervous system. Collectively, these changes are the basis for the many forms of hyperalgesia that can present clinically as incident pain. Knowledge of the various types of hyperalgesia and their underlying mechanisms is required for better treatment of this challenging aspect of chronic pain. PMID- 11014458 TI - A neuroimmune interaction in painful peripheral neuropathy. AB - BACKGROUND: In almost every neuropathic pain state caused by peripheral nerve damage, whether due to trauma or disease, both structural damage and an inflammatory response exist. OBJECTIVE: The goal of this study was to determine the contribution, separate from the effects of the structural lesion, of the inflammatory response to neuropathic pain. METHODS: Two sets of experiments were performed. In the first, an experimental inflammation of the nerve (a neuritis) and an experimental inflammation of the muscle (a myositis control group) were produced in two groups of rats. The pain responses to stimuli applied to the sciatic nerve territory on the plantar hind paw were evaluated through tests of (1) heat hyperalgesia; (2) mechanical allodynia; (3) mechanical hyperalgesia; and (4) cold allodynia. In the second set of experiments, thalidomide or cyclosporin A was used to block the production of immune modulators in the neuritis model and in a chronic constriction injury model (which involves structural damage and an inflammatory response in the sciatic nerve) to determine the contribution of the immune response to the pain observed in the first set of experiments. RESULTS: In experiment 1, rats with the neuritis but not those with the myositis developed neuropathic pain symptoms. In experiment 2, thalidomide produced a partial but significant reduction in pain in the chronic constriction injury model across all four tests, but there was no effect in the neuritis model in any of the tests. Cyclosporin-A resulted in a dose-related reduction in pain in both models across all four tests. CONCLUSIONS: These data suggest the possibility of an important interaction between the immune system and the nervous system in neuropathic pain and suggest that drugs modulating the immune system may be useful therapies in at least some neuropathic pain states. PMID- 11014459 TI - Pain mechanisms and management: a central perspective. AB - Although pain is always intense and unpleasant, the capacity to experience this sensation is, under normal circumstances, fundamental to the preservation of bodily integrity. Clinically, however, after injury to peripheral tissue or directly to the nervous system, spontaneous and evoked pain manifest that serve no physiologic function, are crippling to patients, and are difficult to treat. Here, we review the specific role of the dorsal horn of the spinal cord in the mechanisms of nociceptive protective pain and the spinal plasticity that occurs after nerve and tissue injury. This spinal neuronal plasticity is shown to be a key contributor to pathologic pain hypersensitivity. The potential for the molecular mechanisms responsible for the spinal plasticity in revealing new targets for future treatment is also discussed. PMID- 11014460 TI - Expanded use of surfactant replacement therapy. AB - There are a number of respiratory diseases affecting infants in which there is surfactant dysfunction or deficiency. Surfactant is inactivated by cholesterol, free fatty acids and bilirubin in meconium aspiration syndrome, by haemoglobin and red blood cell lipids in pulmonary haemorrhage and plasma proteins are the culprit in conditions associated with increased alveolar capillary permeability. Surfactant production can be adversely affected by damage to the type 2 pneumocytes by viruses and neutrophil derived reactive oxygen metabolites. Not surprisingly, therefore, the efficacy of exogenous surfactant has been tested, usually in animal models and anecdotal series in "non respiratory distress syndrome" respiratory disorders. Improvements in oxygenation have usually been described. Relatively few randomized trials, however, have been performed, but those undertaken have demonstrated longer term benefits. In term infants with severe respiratory failure, surfactant administration significantly shortened the duration of extracorporeal membrane oxygenation and, in those in the early phase of severe respiratory failure or with meconium aspiration syndrome, it significantly reduced the need for extracorporeal membrane oxygenation. In meconium aspiration syndrome, a smaller number of surfactant treated patients compared to controls developed airleaks. Surfactant administration was also associated with a reduction in the duration of ventilation and intensive care unit stay in patients with meconium aspiration syndrome or bronchiolitis. Those data are very promising and should encourage studies to identify the optimum type of surfactant, dosage regimen and administration method. CONCLUSION: Further randomized trials are required to fully assess the efficacy and cost benefit ratio of surfactant treatment in "non respiratory distress syndrome" respiratory diseases. PMID- 11014461 TI - Reye syndrome revisited: a descriptive term covering a group of heterogeneous disorders. AB - Reye syndrome, characterised by the combination of liver disease and noninflammatory encephalopathy, is a non-specific clinicopathological entity and a descriptive term covering a group of heterogeneous disorders. Nowadays, some of these patients are diagnosed more correctly as having infectious, metabolic, toxic or other disease. The non-specific case definition implies that the epidemiological studies suggesting a link with acetylsalicylic acid have been performed on a heterogeneous group of children, whereby the value of these studies and their ensuing hypothesis is weakened. Moreover, a detailed analysis of the epidemiological surveys of the Centers for Disease Control, the Yale study and of the British risk factor study provides evidence that not only the use of acetylsalicylic acid but also that of phenothiazines and other anti-emetics is significantly greater in Reye syndrome cases than in controls. As to the decline of Reye syndrome, recent literature data reveal that this is related to more accurate modern diagnosis of infectious, metabolic or toxic disease, reducing the percentage of idiopathic or true cases of Reye syndrome. CONCLUSION: Reye syndrome is a non-specific descriptive term covering a group of heterogeneous disorders. Moreover, not only the use of acetylsalicylic acid but also of antiemetics is statistically significant in Reye syndrome cases. Both facts weaken the validity of the epidemiological surveys suggesting a link with acetylsalicylic acid. PMID- 11014462 TI - Current evidence for the use of paediatric antiretroviral therapy--a PENTA analysis. Paediatric European Network for the Treatment of AIDS Steering Committee. AB - The introduction of combination antiretroviral therapy has been associated with a dramatic clinical improvement in children with human immunodeficiency virus infection. However, the uptake of antiretroviral therapy has been variable across Europe. The Paediatric European Network for the Treatment of AIDS Steering Committee has performed a systematic literature review of paediatric antiretroviral therapy trials. An analysis of the evidence base for the commencement and maintenance of antiretroviral therapy was produced. Suggestions for when to commence antiretroviral therapy, which drugs to start with and how to monitor and sequence drug regimens are given. CONCLUSION: The aim of these guidelines is to help in obtaining equity of access to a uniformly high standard of care for children with human immunodeficiency virus infection in all European countries. PMID- 11014463 TI - A preterm baby with Omenn syndrome. AB - A preterm baby born with scaly skin who later developed recurrent infections and was subsequently diagnosed to have Omenn syndrome is presented. CONCLUSION: Any baby with ichthyotic skin and recurrent infections should have immunodeficiency considered in the differential diagnosis. PMID- 11014464 TI - Diagnostic potential of neutrophil elastase inhibitor complex in the routine care of critically ill newborn infants. AB - It has been suggested that determination of the neutrophil elastase alpha1 proteinase inhibitor complex (E-alpha1PI) improves the diagnosis of bacterial infection in newborns. We evaluated the use of E-alpha1PI measurements in 143 newborns, consecutively admitted to a tertiary intensive care unit, employing a new random access assay and a sampling procedure that minimises post-collection artefacts. The 95% range for noninfected newborns was 20-110 microg/l up to the 5th day of life and 20-85 microg/l thereafter. The sensitivity as to the diagnosis of culture-proven bloodstream infection was 80% for E-alpha1PI, 86% for the immature to total neutrophil ratio, 64% for C-reactive protein and 37% for the total white blood cell count. The corresponding specificity amounted to 97%, 85%, 85% and 86%, respectively. E-alpha1PI increases preceded elevations of C reactive protein by 18 h. Like C-reactive protein, E-alpha1PI levels did not distinguish between bloodstream infection and non-bacterial inflammatory responses. Results of E-alpha1PI became available within 1 h of collection and usually 2-3 h before manual leucocyte counts. CONCLUSION: Determination of neutrophil elastase alpha1-proteinase inhibitor levels yields diagnostic advantages comparable to those of manual differential counts but provide faster turnaround times. PMID- 11014465 TI - Risk factors and determinants of neurodevelopmental outcome in cystic periventricular leucomalacia. AB - The aim of the study was to determine risk factors for the development of cystic periventricular leucomalacia (PVL) and to correlate ultrasound findings with neurodevelopmental outcome. By means of a retrospective case-control study (matched for gestational age, birth weight, sex, and year of birth) and a cohort analysis of all preterm infants with cystic PVL documented by ultrasound scans hospitalised at a local tertiary care centre between 1988 and 1998, 98 preterm infants with a gestational age ranging from 26 to 35 weeks were diagnosed as having cystic PVL. The mean day of diagnosis of periventricular echodensities was 3 +/- 2 days (range 1-11 days), and of cystic PVL 21 +/- 8 days (range 2-47 days). Of 79 infants (1988-1997) eligible for neurodevelopmental follow-up (91%), hemi-, di-, or tetraplegia was diagnosed in 61 (77%), normal mental outcome in 22 (28%), associated visual disorders in 41 (52%) and seizure disorders in 12 (15%) infants. Significant risk factors associated with the development of cystic PVL were premature rupture of membranes, chorioamnionitis, and hyperbilirubinaemia (odds ratios 4.665, 6.026, and 2.460 respectively). Subgroup analysis according to gestational age (26-28, 29-32, 33-35 weeks) revealed similar results despite spontaneous labour (26-28 weeks; odds ratio 4.808) and pre-eclampsia (33-35 weeks; odds ratio 3.517). Multiple pregnancy was associated with a twofold increased risk (odds ratio 2.075). The white matter damage probably accounted for the significantly higher prevalence of apnoeas (P < 0.001) and neonatal seizures (P < 0.001). Cysts located bilateral or parieto-occipital were associated with a higher risk of cerebral palsy (odds ratios 6.933 and 4.327 respectively). Solely anterior located cysts were associated with normal neurological outcome. Increasing size of the cysts was associated with increasing risk of cerebral palsy with a cut-off value of 10 mm (odds ratio 3.300 and above) and all infants with cysts of more than 20 mm diameter had cerebral palsy. CONCLUSION: The high prevalence of premature rupture of the membranes and chorioamnionitis further supports the role of intra-uterine infection in the pathogenesis of periventricular leucomalacia. The overall prognosis of cystic periventricular leucomalacia is poor. PMID- 11014466 TI - Prepubertal diagnosis of X-linked congenital adrenal hypoplasia presenting after infancy. AB - X-linked congenital adrenal hypoplasia (CAH) presents classically with adrenal insufficiency within the first 6 months of life, as the fetal adrenal cortex progressively involutes. However, there is increasing recognition of delayed presentation after infancy with the need for accurate molecular diagnosis to avoid an erroneous diagnosis of other more common causes of adrenal insufficiency in childhood. We report our genetic studies of a pedigree with two affected boys presenting with late onset X-linked CAH, diagnosed by the presence of a known W171X mutation of the DAX-1 gene, in whom the mother was an obligate heterozygote. Unlike other causes of adrenal insufficiency, the significance of this diagnosis lies in the important association of hypogonadotropic hypogonadism, and the provision of accurate genetic counselling. CONCLUSION: This study demonstrates that genetic analysis for X-linked congenital adrenal hypoplasia is essential to confirm the diagnosis in prepubertal patients presenting with adrenal insufficiency after infancy. PMID- 11014467 TI - Successful treatment of bilateral renal fungal balls with liposomal amphotericin B and fluconazole in an extremely low birth weight infant. AB - At the age of 8 weeks, an extremely low birth weight infant (gestational age 26 0/7 weeks, birth weight 740 g) had non-obstructing bilateral renal fungal balls. Urine cultures had repeatedly grown Candida albicans. Combination therapy with liposomal amphotericin B intravenously and fluconazole orally was administered for 6 weeks. Monotherapy with fluconazole was then continued until complete resolution of the renal fungal balls. CONCLUSION: Combination therapy with liposomal amphotericin B and fluconazole was successful in eliminating non obstructing bilateral renal fungal balls and obviated the need for surgical intervention. PMID- 11014468 TI - Hospital stay and short-term follow-up of children of drug-abusing mothers born in an urban community hospital--a retrospective review. AB - In order to assess the current use of medical and social services of children of drug-abusing mothers in regard to their short term outcome in a Swiss urban community hospital, we compared hospital, private paediatricians and home nursing records of 37 of these children with 37 matched control children from birth to 18 months of age. Children of drug-abusing mothers (CDAM) experienced a longer neonatal hospital stay than control children with a median (25%-75%) of 26 days (10.5-52.5 days) versus 5 (5-6) days (P < 0.001), a substantial part of which, 8 days (3.5-26 days) versus 0 days (0-1 day) (P < 0.001) was not motivated by any specific medical treatment or nursing care. Before discharge, CDAM were referred to out of hospital nursing and social services for further management, but only 13% were effectively followed. More than 50% were lost to follow-up by their initial paediatrician after 1 year of life. CONCLUSION: New ways to ensure better co-ordination between paediatricians and the social services (inside and outside the hospital) should be developed to shorten the neonatal hospitalisation period and improve the quality of follow-up. PMID- 11014469 TI - Necrotising pneumonitis in children. AB - We retrospectively analysed the clinical features and outcome of children under 17 years of age with necrotising pneumonitis (NP). The radiographs and CT scans of the chest of children under 17 years of age between July 1995 and March 1999 who had complicating community-acquired pneumonia were reviewed. CT scans were obtained for persistent fever, respiratory distress and sepsis despite empiric antibiotic therapy and closed tube drainage. A total of 21 children had the radiographic features of NP of whom 11 (52%) patients were successfully managed using antibiotic therapy with or without closed tube drainage. Ten patients required thoracoscopic decortications and/or lysis of pleural adhesions or debridement of empyema due to refractory pleural sepsis, failure of pulmonary re expansion and persistent air-leaks. The most common pathogens identified were Streptococcus pneumoniae (n = 3), Staphylococcus aureus (n = 2), and Haemophilus influenzae type b (n = 2). The days of hospital stay, duration of fever and days of C-reactive protein return to normal were significantly less for the medically versus the surgically treated children (P < 0.05). CONCLUSION: The clinical course of necrotising pneumonitis in children following complicated pneumonia is often prolonged despite adequate antibiotic therapy. Necrotising pneumonitis with co-existing multiple loculations, pneumothorax/ bronchopleural fistula in the empyema and extensive pleural peel are poor prognostic factors for medical therapy. Thoracoscopic removal of loculated empyema, lysis of adhesions and/or decortication are effective in relieving tachypnoea, chest pain, and controlling fever and improve the outcome, especially in children with empyema. PMID- 11014470 TI - Staphylococcus aureus septicaemia in a patient with cystic fibrosis. AB - Although bacterial colonisation of bronchi may occur from early childhood onwards, infections extending beyond the lungs are uncommon in patients with cystic fibrosis. A 12-year-old boy with cystic fibrosis, receiving oral corticosteroids for 3 weeks because of allergic bronchopulmonary aspergillosis, experienced pneumonia and septicaemia caused by Staphylococcus aureus. He was treated with flucloxacillin, ticarcillin-clavulanate, aztreonam, cefazolin and rifampin according to resistance testing of S. aureus cultured from the blood. On day 25 the patient finally had recovered. CONCLUSION: Systemic steroid therapy for allergic bronchopulmonary aspergillosis may favour life-threatening systemic bacterial infection which is rare in the immunocompetent patient with cystic fibrosis. PMID- 11014471 TI - When do children with optic pathway tumours need treatment? An oncological perspective in 106 patients treated in a single centre. AB - Progression patterns of optic pathway tumours (OPT) need to be precisely defined for treatment planning. In patients with neurofibromatosis type 1 (NF1), this disease is usually indolent and the available literature rarely reports progression after the age of 6 years. In patients without NF1, the disease course seems to be less favourable. We reviewed the clinical and radiological files of 106 children referred to our institution for the treatment of a symptomatic OPT since 1980. NF1 was present in 51 of them. Progression patterns in children with NF1 differed markedly from those in the other patients. A total of 83 children had tumour extension beyond the chiasm (Dodge type III). Children with NF1 had progressive tumours later during follow-up (47% after the age of 6 years), had more often proptosis and infiltrating tumours but less frequently nystagmus or increased intracranial pressure. 32 children were not treated at diagnosis because they had only mild symptoms related to the OPT. In these patients, progression occurred more often in children without than with NF1 (12/12 versus 12/20 respectively, P = 0.04). A high number of patients needed treatment for progression or severe symptoms after 6 years of age. Of the patients, 33% needed treatment for progression or severe symptoms after 6 years of age. CONCLUSION: Progression patterns of optic pathway tumours in children with neurofibromatosis type 1 differ markedly from those in other patients. This study emphasises the need for prolonged follow-up of children with optic pathway tumours, especially in neurofibromatosis type 1. PMID- 11014473 TI - Immunodeficiency in alpha-mannosidosis: a matched case-control study on immunoglobulins, complement factors, receptor density, phagocytosis and intracellular killing in leucocytes. AB - Patients with the autosomal recessive lysosomal storage disease alpha mannosidosis suffer from recurrent infections. To study the mechanisms of this immunodeficiency, six patients were matched against six healthy controls and their humoral and cellular immunocompetence investigated. No differences in the number of circulating leucocytes including B-cells, levels of immunoglobulin main classes, nor IgG subclasses were observed. However, post-immunisation serum levels of specific antibodies against poliovirus, diphtheria toxin and tetanus toxin were significantly reduced. In patients, the density of the complement binding receptor CD11b and the Fc-receptor CD16 was significantly enhanced on monocytes and polymorphonuclear neutrophils (PMN) and the number of phagocytosing PMN was significantly increased in the presence of pooled human serum. This was not observed in the presence of autologous serum, indicating altered opsonic properties. Also in normal PMN, phagocytosis was inhibited by a factor in the serum from the patients. Despite maintained oxidative burst, patient PMN demonstrated insufficient intracellular bacterial killing. CONCLUSION: Our data indicate that patients with alpha-mannosidosis have an immunodeficiency at both the humoral and cellular level. PMID- 11014472 TI - Therapeutic drug monitoring of once daily gentamicin in serum and saliva of children. AB - Gentamicin is widely used in paediatric medicine and therapeutic monitoring is mandatory due to the narrow margin of safety. Saliva sampling may be of potential interest, especially in children in whom blood sampling is often difficult. Experience with once daily intravenous administration of aminoglycosides has grown in recent years. Gentamicin levels were measured in serum and saliva of 55 children treated with the drug (5 mg/kg per day), administered intravenously in three different regimens: thrice (n = 19), twice (n = 18), and once daily (n = 18). No correlation was found between serum gentamicin concentrations and saliva levels when the drug was administered twice or thrice daily, however, there was good correlation when the drug was administered once daily (r2 = 0.96, P < 0.0001). CONCLUSION: In children with uncomplicated infections treated with once daily gentamicin, trough concentrations of the drug can be monitored in saliva. PMID- 11014474 TI - Acute disseminated encephalomyelitis--a diagnosis to consider. AB - The case history of an 11-year-old boy with post-infectious acute disseminated encephalomyelitis (ADEM) is presented. ADEM is a diagnosis that is increasingly being made with the advent of modern radiological techniques. However, the syndrome cannot be thought of as a discrete clinical entity as it is triggered by a wide range of factors including infections and vaccinations. Furthermore, other pathological processes may give the same radiological appearances. The aetiology, pathophysiology and management of the syndrome are discussed. CONCLUSION: Acute disseminated encephalomyelitis is a radiological diagnosis that has many precipitants. Thought must be applied in developing a differential diagnosis to allow the possibility of targeting treatment at the underlying cause. PMID- 11014475 TI - An infant with multiple soft tissues masses and abnormal radiological bone anomalies. Infantile myofibromatosis. PMID- 11014476 TI - Cerebral vasculitis stabilised by methotrexate. PMID- 11014477 TI - Interleukin-2 mediated restoration of natural killer cell function in a patient with Griscelli syndrome. PMID- 11014478 TI - Colonic perforation in two children with Campylobacter enterocolitis. PMID- 11014479 TI - Transient neonatal hypothyroidism during breastfeeding after post-natal maternal topical iodine treatment. PMID- 11014480 TI - Cross-sectional atomic force microscopy imaging of polycrystalline thin films AB - Atomic force microscopy (AFM) can be used to image cross-sections of thin-film samples. So far, however, it has mainly been used to study cross-sections of epitaxial systems or integrated circuits on crystalline substrates. In this paper, we show that AFM is a powerful tool to image fractured cross-sections of polycrystalline thin films deposited on crystalline and non-crystalline substrates, yielding unique information on the three-dimensional properties of the cross-sections, with a spatial resolution in the nm range. Original images of three different heterostructure systems are presented: Si(wafer)/SnO2/CdS/CdTe, glass/Mo/Cu(In,Ga)Se2,/CdS/ZnO, and glass/SnO2/WO3. We discuss the results by comparing AFM and scanning electron microscopy (SEM) images, and explain, for the different materials, why the AFM provides useful additional information. PMID- 11014481 TI - A new phase consistency criterion and its application in electron crystallography AB - In this work, we present the principles and potential advantages of a methodology to assess Fourier components in terms of phase consistency. We define a new phase consistency criterion among sets of spatially translated images based upon a novel conception of the spatial shift property of the Fourier transform. The article shows how this criterion can be used in the alignment stage of the 3D reconstruction process with a two-fold objective: Assessment of the frequency components and robustness in the alignment. In that sense, the article shows and analyzes the results obtained from the application of the new index of quality in the context of projection image alignment. We have focussed our attention on the electron crystallography field, by applying such a phase consistency definition over image reflections. The results that have been obtained show that the new phase-consistency definition may complement the traditional SNR-based index of quality (commonly known as IQ) of reflections. As a consequence, the reliability of the alignment may be improved by discarding those reflections judged as non reliable according to the phase-consistency criterion. PMID- 11014482 TI - Selective specimen preparation for TEM observation of the cross-section of individual carbon nanotube/metal junctions AB - We present here an efficient method to prepare a transmission electron microscopy (TEM) specimen for selective observation of the cross-section of individual nanoscale structures. As a typical example, the cross-sectional TEM observation of a quasi-one-dimensional material - a nano-electronic component based on an individual carbon nanotube - is presented. PMID- 11014483 TI - Alignment of AFM images using an iterative mathematical procedure AB - An iterative mathematical procedure for the alignment of sequentially recorded atomic force microscope images (AFM) is presented. The computer program is able to correct commonly observed drifts in vertical and lateral directions, rotations around a vertical or lateral axis and differences in scale. This method is applied on dissolution experiments of uranium dioxide (UO2) surfaces. Images recorded during in situ experiments, which are shifted probably due to thermal fluctuations, can be aligned with good accurancy. In a further approach the UO2 surface is marked by electron-beam-induced deposition (EBD or EBID) with microstructured reference points. The alignment can be distinctly improved using marked sample surfaces because of the characteristic shape of the markings, which do not change during the experiment. Furthermore, the markings can be used to find again a domain on a sample surface. Therefore, AFM images recorded before and after an ex situ experiment (e.g. treatment in corrosive medium for a longer period of time) can be aligned with a nanometer spatial resolution. PMID- 11014484 TI - Light atom derivatives of structure-preserving sugars are unconventional negative stains. AB - Although glucose and certain other sugars are known to greatly reduce distortion and denaturation of proteins during drying, use of this monosaccharide as an experimental negative stain does not permit imaging of lattice periodicities in test specimens of thin catalase crystals. However, the potassium and sodium salts of several forms of monophosphorylated glucose (200 mM), diphosphorylated glucose, monosulfated glucose, maltose-1-phosphate, and trehalose-6-phosphate, all dry into a glassy layer and scatter transmitted electrons sufficiently to show the 86 A major periods in catalase crystals. Glucose-6-phosphate provides sufficient image contrast at concentrations from 2 mM (=0.067%) to 500 mM (= 16.8%). Underfocusing increases visualization of the periodic lattice, indicating a large contribution of phase contrast to these images. Upon exposure to the electron beam, thicker regions of derivatized saccharides or pure glucose develop bubbling; this redistribution of dried stain largely can be precluded by imaging with low-dose exposures. Power spectra of images of catalase crystals contained within 200 mM disodium glucose-6-phosphate show that periodic information can be recorded to 21 A; some individual features of dipotassium glucose-6-phosphate distribution within the protein lattice have a measured width of around 5 A. The experimental results demonstrate that structure-preserving mono- and di saccharides also serve successfully as negative stains after they are coupled to light atom scatterers. PMID- 11014485 TI - Cross-sectional study of the prevalence of feather pecking in laying hens in alternative systems and its associations with management and disease. AB - A cross-sectional study of risk factors for feather pecking in layings hens in alternative systems was carried out in July 1998. A total of 637 questionnaires were sent out to farmers and producer groups and, after two reminders, the final response rate was 51.5 per cent. The outcome variable was feather pecking after point of lay. Over 55 per cent of the farmers reported that feather pecking had occurred in the last depopulated flock. This outcome was compared with the management procedures reported by flock managers by using univariate statistics. Factors associated with feather pecking with a significance < or = 0.05 were then tested in two logistic regression models. In the first model the following factors were associated with an increased risk of feather pecking: less than 50 per cent of the flock using the outdoor area on a fine and sunny day, the occurrence of egg peritonitis and the occurrence of infectious bronchitis. The direction of the association between feather pecking and these infectious diseases was unclear, so in the second model only factors which were consistent throughout the laying period were tested. The following factors were associated with an increased risk of feather pecking: less than 50 per cent of the flock using the outdoor area on a fine and sunny day; three or more changes of diet during lay; the inspection of the flock by one person; an absence of loose litter at the end of lay; a temperature in the hen house of less than 20 degrees C; turning the lights up when the flock was inspected; and the use of bell-drinkers. It is concluded that some of these factors could inhibit foraging and dust bathing behaviour and others may increase competition or frustration, both of these changes having been shown experimentally to initiate feather pecking behaviour. PMID- 11014486 TI - Fatal adiaspiromycosis in a wild Eurasian otter (Lutra lutra). AB - Postmortem examination of an immature otter which died in the wild showed that large areas of the lungs were swollen and firm, with emphysema and haemorrhage in the remaining areas. Histopathological examination revealed large numbers of fungal adiaspores and an unusually severe inflammatory response. It was considered that respiratory impairment was the primary cause of the otter's death. PMID- 11014487 TI - Surgical treatment of coenurosis (gid) in sheep. AB - During the past six years 623 cases of coenurosis (gid) in sheep have been treated surgically. Cysts were removed successfully from 573 of them (92 per cent) and 517 (83 per cent) were able to return to their flocks, although 36 showed no clinical improvement In 37 cases, the cyst could not be localised, and postmortem examinations showed that in nine cases the cyst was in the brainstem, and in 28 cases it was in the cerebellum. Fifty-six cases deteriorated gradually after surgery and in these cases more than one cyst was found postmortem. Thirteen cases died during surgery. PMID- 11014488 TI - Spring survey of the parasite Heterakis gallinarum in wild-living pheasants in Britain. PMID- 11014489 TI - First isolations of leptospires serogroup Ballum serovar arborea in Argentina. PMID- 11014490 TI - Spontaneous resolution of splenic torsion in a dog. PMID- 11014491 TI - CSF virus in East Anglia: where from? PMID- 11014492 TI - Hunting inquiry. PMID- 11014494 TI - Dermal fibrosis in a rabbit. PMID- 11014493 TI - Winter transmission of equine nematodes. PMID- 11014495 TI - Prostaglandin use in sows. PMID- 11014496 TI - Computer viruses, worms and Internet security. PMID- 11014497 TI - Sperm binding and penetration of the zona pellucida in vitro but not sperm-egg fusion in an Australian marsupial, the brushtail possum (Trichosurus vulpecula). AB - Sperm capacitation and in vitro fertilisation (IVF) have been achieved in most eutherian mammals and American marsupials under relatively simple culture conditions. In contrast sperm capacitation in Australian marsupials has not been achieved in vitro and attempts at IVF have previously been characterised by a complete lack of sperm-zona pellucida (ZP) binding. Recently, co-culture of sperm with oviduct epithelial cell monolayers or with oviductal explant conditioned media has been shown to prolong the viability and motility of brushtail possum spermatozoa, as well as to induce capacitation-associated changes such as transformation of sperm to the T-shape orientation. In this study we report that these in vitro produced T-shaped sperm, and in vivo derived T-shaped sperm flushed from the oviduct of artificially inseminated possums as a control, are able to bind to and penetrate the ZP of approximately 25% of eggs recovered from PMSG/LH-superovulated possums in vitro. Development of ZP receptivity and penetrability towards sperm was also identified as a major factor affecting the outcome of IVF. Neither in vivo nor in vitro derived T-shaped sperm were able to bind to or penetrate the ZP if eggs were obtained from animals that were treated with pLH less than 76 h after PMSG. Thus this study provides preliminary evidence for the necessity of sperm-oviduct epithelial cell interactions for capacitation in Australian species and lends further support to the suggestion that the T shape head orientation is indicative of sperm capacitation. Despite the occurrence of sperm-ZP binding and penetration, sperm-egg membrane fusion and egg activation were not observed. Although the factor(s) responsible for the lack of sperm-egg membrane fusion in the possum have not been identified it is possible that egg capacity for membrane fusion develops independently of zona receptivity and is defective in these eggs, or alternatively that membrane fusion requires strictly defined ionic conditions which are not provided by the IVF media used in this study. PMID- 11014498 TI - Sperm decondensation during fertilisation in the mouse: presence of DNase I hypersensitive sites in situ and a putative role for topoisomerase II. AB - In this study our aim was to characterise the presence and the role of DNA alterations during sperm decondensation in the mouse. To visualise the changes during decondensation we investigated for the presence of DNase I hypersensitive sites in situ and for a putative role for topoisomerase II by examining the effect of teniposide, a topoisomerase II inhibitor, during fertilisation. In situ nick translation without the previous addition of DNase I failed to reveal the presence of endogenous nicks in decondensing sperm and pronuclei whereas preincubation of fixed oocytes with DNase I indicated that decondensing sperm were sensitive to this enzyme. Addition of 100 microM teniposide did not completely inhibit pronuclei formation but its addition to the fertilisation medium did lead to the presence of endogenous DNA nicks in decondensing sperm. These observations suggest that DNase I hypersensitivity during sperm decondensation is related to the dramatic conformational changes that the chromatin undergoes during the decondensation process, in which topoisomerase II may be implicated. PMID- 11014499 TI - Effective activation method with A23187 and puromycin to produce haploid parthenogenones from freshly ovulated mouse oocytes. AB - Freshly ovulated mouse oocytes exposed to 5 mM calcium ionophore A23187 for 5 min and controls (not exposed) were cultured in TYH medium with 10 microg/ml puromycin (the puromycin group) or 2 mM 6-dimethylaminopurine (DMAP; the DMAP group) for 4 h. Among the controls, few oocytes were activated even if they were treated with DMAP or puromycin. In the oocytes exposed to A23187, in contrast, the activation rate, i.e. the rate of oocytes showing at least one pronucleus (PN) after the treatment, was 46.2% (48/104) in the DMAP group and 90.0% (118/131) in the puromycin group. Activation rate in the puromycin group was significantly higher than in the DMAP and control groups (p < 0.0001, respectively). Furthermore, 82.4% (108/131) of the activated oocytes in the puromycin group showed one PN with extrusion of the second polar body (PB). In the puromycin group, the DNA content of the PN of parthenogenones with 1PN2PB was half that of a set of metaphase II chromosomes. Chromosomal analysis was possible in 14 parthenogenones with 1PN2PB in the puromycin group. The parthenogenones possessed a normal set (n = 20) of haploid chromosomes. The combination of A23187 and puromycin proved to be an effective method of producing haploid parthenogenones. PMID- 11014500 TI - Quantification of mtDNA in single oocytes, polar bodies and subcellular components by real-time rapid cycle fluorescence monitored PCR. AB - Oocytes, in general, are greatly enriched in mitochondria to support higher rates of macromolecular synthesis and critical physiological processes characteristic of early development. An inability of these organelles to amplify and/or to accumulate ATP has been linked to developmental abnormality or arrest. The number of mitochondrial genomes present in mature mouse and human metaphase II oocytes was estimated by fluorescent rapid cycle DNA amplification, which is a highly sensitive technique ideally suited to quantitative mitochondrial DNA (mtDNA) analysis in individual cells. A considerable degree of variability was observed between individual samples. An overall average of 1.59 x 10(5) and 3.14 x 10(5) mtDNA molecules were detected per mouse and human oocyte, respectively. Furthermore, the mtDNA copy number was examined in polar bodies and contrasted with the concentration in their corresponding oocytes. In addition, the density of mtDNA in a cytoplasmic sample was estimated in an attempt to determine the approximate number of mitochondria transferred during clinical cytoplasmic donation procedures as well as to develop a clinical tool for the assessment and selection of oocytes during in vitro fertilisation procedures. However, no correlation was identified between the mtDNA concentration in either polar bodies or cytoplasmic samples and their corresponding oocyte. PMID- 11014501 TI - Lack of cell cycle checkpoints in human cleavage stage embryos revealed by a clonal pattern of chromosomal mosaicism analysed by sequential multicolour FISH. AB - Multicolour fluorescence in situ hybridisation (FISH) analysis of interphase nuclei in cleavage stage human embryos has highlighted a high incidence of postzygotic chromosomal mosaicism, including both aneuploid and ploidy mosaicism. Indeed, some embryos appear to have a chaotic chromosomal complement in a majority of nuclei, suggesting that cell cycle checkpoints may not operate in early cleavage. Most of these studies, however, have only analysed a limited number of chromosomes (3-5), making it difficult to distinguish FISH artefacts from true aneuploidy. We now report analysis of 11 chromosomes in five sequential hybridisations with standard combinations of two or three probes and minimal loss of hybridisation efficiency. Analysis of a series of arrested human embryos revealed a generally consistent pattern of hybridisation on which was superimposed frequent deletion of one or both chromosomes of a specific pair in two or more nuclei indicating a clonal origin and continued cleavage following chromosome loss. With a binucleate cell in a predominantly triploid XXX embryo, the two nuclei remained attached during preparation and the chaotic diploid/triphoid status of every chromosome analysed was the same for each nucleus. Furthermore, in each hybridisation the signals were distributed as a mirror-image about the plane of attachment, indicating premature decondensation during anaphase consistent with a lack of checkpoint control. PMID- 11014502 TI - Effects of sex steroid hormones and their antagonists on mast cell number in the testis of the frog, Rana esculenta. AB - This study confirms our previous data on the effects of sex hormones on mast cell number (MCN) in the testis of frog Rana esculenta. After 15 days of treatment with oestradiol (E2) MCN strongly increases, while testosterone has no effect. After 30 days only a small increase in MCN is observed. These differences could be due to the non-physiological effect of E2 over a prolonged period. We also confirmed a massive increase in MCN after 15 or 30 days of treatment with cyproterone acetate (CPA). This increase in MCN is also observed after administration of CPA with tamoxifen. Ultrastructural analysis of testis shows empty spaces with degenerating Leydig cells in the interstitial compartment and numerous germinal cells completely degenerated, probably apoptotic, in the adjacent germinal compartment. The same effects were observed in testes after treatment with only CPA. Chronic E2 treatment provokes an increase in MCN on day 2. From day 4 to 12 of the treatment, MCN decreases dramatically and many germinal tubules appear strongly disorganised. In conclusion, the present results confirm that E2 treatment induces changes in MCN and chronic E2 treatment modifies the morphology of the frog testes. In addition, blocking androgen receptors with CPA, alone or in combination with tamoxifen, causes a significant increase in MCN, confirming the involvement of androgens in mast cell proliferation and/or differentiation. PMID- 11014503 TI - The effects of embryo stage and cell number on the composition of mouse aggregation chimaeras. AB - Studies with intact preimplantation mouse embryos and some types of chimaeric aggregates have shown that the most advanced cells are preferentially allocated to the inner cell mass (ICM) rather than the trophectoderm. Thus, differences between 4-cell and 8-cell stage embryos could contribute to the tendency for tetraploid cells to colonise the trophectoderm more readily than the ICM in 4 cell tetraploid<-->8 cell diploid chimaeras. The aim of the present study was to test whether 4-cell stage embryos in 4-cell diploid<-->8-cell diploid aggregates contributed equally to all lineages present in the E12.5 conceptus. These chimaeras were compared with those produced from standard aggregates of two whole 8-cell embryos and aggregates of half an 8-cell embryo with a whole 8-cell embryo. As expected, the overall contribution of 4-cell embryos was lower than that of 8-cell embryos and similar to that of half 8-cell stage embryos. In the 4 cell<-->8-cell chimaeras the 4-cell stage embryos did not contribute more to the trophectoderm than the ICM derivatives. Thus, differences between 4-cell and 8 cell embryos cannot explain the restricted tissue distribution of tetraploid cells previously reported for 4-cell tetraploid<-->8-cell diploid chimaeras. It is suggested that cells from the more advanced embryo are more likely to contribute to the ICM but, for technical reasons, are prevented from doing so in simple aggregates of equal numbers of whole 4-cell and whole 8-cell stage embryos. PMID- 11014504 TI - Chlortetracycline staining patterns of frozen-thawed bull spermatozoa treated with beta-cyclodextrins, dibutyryl cAMP and progesterone. AB - Efforts to achieve complete chemical definition of media used for in vitro capacitation of bovine spermatozoa including removal of heparin purified from porcine intestinal mucosa are presented. Fluorescent staining with chlortetracycline (CTC), known to reflect changes coincident with sperm capacitation in certain species, was studied following treatments of frozen thawed bull spermatozoa with beta-cyclodextrins, dibutyryl cAMP (dbcAMP) and progesterone in comparison with heparin. The CTC staining patterns (F, B and AR) were confirmed to correlate with known conditions that effectively prepare cryopreserved bull spermatozoa for fertilisation in vitro. In the absence of glucose, the routinely employed heparin-containing capacitating medium caused an increase in spermatozoa displaying the AR pattern. Both progesterone (100 microM) and dbcAMP (0.01-0.1 mM) were able to increase the proportion of B pattern stained sperm cells more than after exposure to control (mDM) conditions without a significant reduction in motility. Exposure to either dbcAMP or beta cyclodextrins was accompanied by an increase in proportions of spermatozoa displaying the AR pattern over those seen in controls. Exposure to beta cyclodextrins did not increase the proportion of B pattern stained spermatozoa. Comparison of spermatozoa from two bulls revealed differential responses of spermatozoa from different males to treatments with heparin and progesterone. In vitro fertilisation results demonstrated that previously cryopreserved bull spermatozoa could be capacitated in chemically defined conditions devoid of heparin or other biological components. PMID- 11014505 TI - Enzymes responsible for the bactericidal effect in extracts of vitelline and fertilisation envelopes of rainbow trout eggs. AB - Extracts from both the vitelline envelope (VE) and fertilisation envelopes (FE) of rainbow trout eggs have the ability to exert a bactericidal effect on Gram positive and -negative bacteria. The effect may be due to the presence of phospholipase D (PLD), lysozyme, proteinase and DNases, as the extracts contain these enzyme activities. The intensity of chorionic PLD and lysozyme activities in the VE extract was maintained in the FE without any alteration in activity even after transformation in the course of the cortical reaction, as components of a fundamental architecture of the envelope. Both extracts also contain different types of proteinase activities. Treatment with VE or FE extract seriously damaged the outer membrane of Gram-negative bacteria and the plasma membrane of Gram-positive and -negative bacteria at the ultrastructural level. Chorionic DNases probably degrade DNA of bacterial cells killed by virtue of the action of PLD and/or lysozyme and contribute to the transmigration of nucleosides and/or nucleotides produced by degrading bacterial DNA after degradation of bacterial components by the actions of the chorionic PLD, lysozyme and proteinase. These results suggest that the bactericidal process manifested by the VE or FE extract may start with the action of PLD and/or lysozyme against bacteria and be completed by subsequent degradation of constitutive proteins and DNA by the action of proteinases and DNases, respectively. Thus the VE and FE are able to protect the egg itself and the embryo, respectively, from bacterial infection in the internal or external environments. PMID- 11014506 TI - Fertilisability of ovine, bovine or minke whale (Balaenoptera acutorostrata) spermatozoa intracytoplasmically injected into bovine oocytes. AB - This study was conducted to investigate the possibility of using bovine oocytes for a heterologous fertility test by intracytoplasmic sperm injection (ICSI) and to compare the pronuclear formation of ram, bull and minke whale spermatozoa after injection into bovine oocytes. Bovine oocytes were cultured in vitro for 24 h and those with a polar body were selected for ICSI. Frozen-thawed semen from the three species were treated with 5 mM dithiothreitol for 1 h and spermatozoa were killed by storing them in a -20 degrees C refrigerator before use. ICSI was performed using a Piezo system. Three experiments were designed. In experiment 1, a higher (p < 0.05) male pronuclear formation rate was found in the oocytes injected with ram (52.6%) or bull (53.4%) spermatozoa than with minke whale spermatozoa (39.1%). In experiment 2, sperm head decondensation was detected at 2 h after ICSI in the oocytes injected with a spermatozoon of each species. Male pronuclei were first observed at 4 h in the oocytes injected with ram or bull spermatozoa and at 6 h in oocytes injected with minke whale spermatozoa. The mean diameters of male pronuclei derived from both whale and bull spermatozoa were larger than those from ram spermatozoa (30.4 microm and 28.3 microm vs 22.4 microm, p < 0.005). The mean diameter of female pronuclei in the oocytes injected with whale spermatozoa was also larger than with ram spermatozoa (29.3 microm vs 24.7 microm, p < 0.05). The development of male and female pronuclei was synchronous. In experiment 3, ethanol-activated oocytes injected with a spermatozoon from any of the three species achieved significantly higher (p < 0.05-0.001) cleavage rates than control oocytes. Blastocyst formation was only observed when bull spermatozoa were used. The results of this study indicate that dead foreign spermatozoa can participate in fertilisation activities in bovine oocytes after ICSI. PMID- 11014507 TI - The effect of growth hormone on rat pre-antral follicles in vitro. AB - The aim of the present study was to investigate whether growth hormone (GH) has any effect on the development of cultured rat pre-antral follicles. Pre-antral follicles with a diameter between 120 microm and 160 microm were mechanically isolated from 10-day-old rat ovaries and cultured in groups for 6 days in serum free medium without GH or with GH supplemented at concentrations of 1, 10 and 100 ng/ml, respectively. DNA content of the follicles before and after culture was measured to determine whether possible growth is due to proliferation of follicular cells. To investigate the quality of follicles cultured under different conditions, the ultrastructure of the cultured follicles was studied with transmission electron microscopy. Furthermore, reverse transcriptase polymerase chain reaction (RT-PCR) was used to assess the expression of growth hormone receptor (GHR) in pre-antral follicles. GH, regardless of the concentration, stimulated the growth of pre-antral follicles. However, follicles cultured in medium supplemented with high-dose GH (100 ng/ml) showed a significantly lower survival rate compared with the other groups. Follicles cultured in GH-containing medium showed a better ultrastructure in comparison with those cultured in medium without GH. Remarkably, scattered cortical granules were observed in oocytes of follicles cultured in the presence of GH. With RT PCR, the presence of the mRNA of GHR was demonstrated in pre-antral follicles. It can be concluded that GH promotes rat pre-antral follicle development in vitro and better supports the morphology of cultured pre-antral follicles. The gene expression of GHR suggests that the action of GH could be mediated by its receptors present in pre-antral follicles. PMID- 11014508 TI - Decision-making for national programs of community fluoride use. AB - Every community, region or country with a high or rising prevalence of dental caries should implement a caries-preventive program that automatically brings the benefits of systemic and topically applied fluoride to the entire population. The fluoridation of community water supplies or salt fulfills the requirements of providing safe, effective protection from dental caries at reasonable cost. The use of dietary fluoride supplements or fluoridated milk does not meet the requirements of a comprehensive national or community program because compliance is poor or only selected age groups are targeted. Water fluoridation is ideal for countries, regions or communities with many central water supplies or where salt production or distribution is not centralized or easy to control. Water fluoridation also has advantages where many areas exist with natural water fluoride concentrations at optimal or greater than optimal concentrations. Salt fluoridation is ideal for countries or regions with few, potable central water supplies in which salt production and distribution are centralized and easily controlled. Concentrations of fluoride for water fluoridation range from 0.5 to 1.2 parts per million (ppm) parts of water depending on climate and dietary practices. The concentration for fluoridation of salt is approximately 200 to 250 mg fluoride per kg of salt, also depending on dietary practices. Properly fluoridated salt should produce levels of urinary fluoride excretion similar to those found in communities with fluoridated water. Benefits of the two methods are similar. Salt fluoridation may be done more cheaply. PMID- 11014509 TI - Reliability and validity of the Dental Indifference Scale in a population of 18 year-olds in Norway. AB - OBJECTIVES: The aims of this study were to estimate the reliability and validity of the Dental Indifference Scale (DIS) (Nuttall, 1996) in a population of 18-yr olds in Norway. METHODS: The DIS-scale was mailed to a sample of 1119 18-yr-olds in two Norwegian counties. Nearly 87% completed the questionnaire and consented to the collection of data from their dental records. Ten percent of the sample, drawn at random, was asked to complete the questionnaire a second time, after a time delay of 15 weeks (response rate 83%). The reliability estimation of the sum scores of DIS was based on Pearson's correlation between test-retest scores and internal consistency (Cronbach's alpha). The frequency of missed appointments from age 12, recorded in the dental treatment records, was used as the validating criterion. The validity was analyzed by Pearson's correlation, and step-wise multiple regression. RESULTS: The correlation coefficient (Pearson) for the test retest comparison was 0.43. The correlation coefficient between the DIS-scores and the frequencies of missed dental appointments was 0.24. The Cronbach's Alpha coefficient for the eight DIS-questions was 0.35 (n=868). Only two of the eight DIS-questions entered the stepwise regression model and explained 15% of the variance of the frequency of missed appointments. CONCLUSIONS: The Dental Indifference Scale (DIS) was found to have a low reliability and validity in this study population, and it is recommended that it should not be used without further investigation. It may be necessary to design an alternative instrument if further work into the hypothesized trait of dental indifference is to be undertaken. PMID- 11014510 TI - Oral hygiene knowledge of high-risk Grade One children: an evaluation of two methods of dental health education. AB - The effectiveness of two methods of dental health education (DHE) for improving oral hygiene knowledge among high-risk Grade One students was evaluated. Fifty elementary schools in the former City of North York, Canada were assigned to one of two groups. In one group, students received a classroom-based DHE lesson which was reinforced by two small-group sessions (n=243). In the other group, students received only a single classroom-based DHE lesson (n=206). After DHE interventions, students in both groups displayed improved knowledge for most oral hygiene questions (e.g., when should you throw your toothbrush away?). However, for several questions, a significantly higher proportion of "classroom plus small group sessions" students displayed improved knowledge compared to students receiving only a classroom lesson. These items included: awareness that cavity prevention and removal of germs are two purposes of oral hygiene; and knowledge that teeth help people to eat and talk. Results suggest a classroom-based lesson combined with small-group sessions is a more effective method of improving oral hygiene knowledge among high-risk Grade One students compared to a single classroom-based lesson. PMID- 11014512 TI - Variation in caries and treatment experience in 35-44-year-old Lithuanians. AB - Data summarising caries levels differ between countries and often conceal large variations. These differences may reflect variations at individual, group, social and other levels. Caries and treatment experience differences are found to be significantly related to a variety of clinical and non-clinical factors. Variation in caries within Lithuanian children has been reported and related to background-, behavior-, dental care- and other factors. Variation in general or related to any factors in adult Lithuanian population has not been studied. The aim of the present study was to estimate variation in caries and treatment experience and relate them to biological-, psychosocial-, health-related, behavioral- and dental care related aspects. The existing knowledge from other oral health studies was used to select the factors found of importance in relation to variations in caries scores. The study consisted of clinical and self reported data collected from a stratified random sample of 382 individuals (attendance rate 50%). Two statistical approaches, a bivariate and a multivariate, were applied. The two dependent variables DMFT and DS were tested with a set of independent variables. The bivariate analysis revealed significant associations for approximately one half of all variables studied. For multivariate testing, a backward linear multiple regression was used. Forty-one percent of the variation in DS was explained and 48% in DMFT scores. Differences in scores were related to various factors in the two analyses. The strength and significance of a few relationships differed in both of the analyses. The multivariate testing revealed some strong and significant associations with independent variables which were found to be weak and non-significant (P>0.05) when tested bivariately and vice versa. Less than half of the variation could be explained in dental caries scores in 35-44-year-olds and not only dental care related factors were important. In both of the analyses, the varying degrees of strength (significance levels) give rise to a hypothesis that this incomplete explanation of the variation can be due to interrelated effects of different estimates. PMID- 11014511 TI - The fractional urinary fluoride excretion in young children under stable fluoride intake conditions. AB - The purpose of this study was to determine the fraction of the total daily fluoride intake that is excreted through the urine (FUEF) of children aged 3-5 years under usual intake conditions. Participating children were residents of an area with a fluoride (F) concentration of 0.5-0.6 mg/L in their drinking water. Assessments were made on two successive 24-h periods on 20 children, measuring the total amount of fluoride ingested through liquid and food consumption, and from ingestion of fluoridated toothpaste (500 microg F/g), together with the determination of the amount of fluoride excreted through urine. Fluoride retention was also estimated assuming a constant average F fraction of 10% excreted through faeces. It was found that the average proportion of liquids, solid foods, and toothpaste to the daily fluoride intake (1.02-mg F/day on average) were 40.8, 34.6, and 24.5%, respectively. The average FUEF value was 35.5% (95% C.I.=31.7-39.3%), and the estimated fractional F retention was 54.5%. The present data suggest a slight relationship between FUEF values and the inverse of the daily fluoride dose (1/dose) (r=0.513; P=0.021). When the present results are combined with those from previous studies on F-retention and urinary excretion, the correlation between both FUEF and fractional retention and 1/dose are very strong and highly significant (r=0.98, P<0.0001, and r=-0.986, P<0.0001, respectively). A possible mechanism is suggested in order to explain this latter finding. The potential usefulness of the current FUEF value for the estimation of daily F intake (or dose) from urinary F excretion data is also discussed. PMID- 11014513 TI - A longitudinal study of young Finnish adults' use of subsidized, private sector dental care, 1986-1997. AB - OBJECTIVES: Finns born after 1956 are now entitled to subsidized private sector dental care, or such persons could be enrolled in the Public Dental Service. Until 1986, eligibility was more restrictive. The purpose of this study was to investigate the use and costs of private dental care and effects of regularity of care on costs and treatment received among young Finnish adults during 1986-1997. METHODS: All 1986, 1990 and 1994 recipients of reimbursement for dental care from the Social Insurance Institution were included in the study. Five separate age cohorts were compared. Using their civil registration numbers, individuals were tracked from their first contact with a private dentist in one of the years 1986, 1990 or 1994 until the year 1997. RESULTS: While the total number of young adults who had received reimbursement for private dental care increased from about 53000 (1986) to 200000 (1994) due to extended eligibility, the number of users in the youngest group decreased from 53000 to 23000. Attending infrequently (1-2 times during the study period) was most common among the youngest adults and frequent attendance (annually) was most common among older adults. The annual mean cost was slightly lower among the frequent attenders in almost every cohort. Variation in the mean number of annual visits was directly correlated with costs. Frequent attenders most often received diagnostic and preventive measures while restorations and surgery were most common for the infrequent attenders. CONCLUSIONS: A substantial decline in the demand for private services among the 19-25-year-olds and stable demand among 26-34-year-olds was detected, indicating falling treatment needs or a preference for the Public Dental Service. All groups had a stable mean number of visits per year and almost constant costs. The mean number of dental visits per year remained steady in all cohorts indicating rigid treatment patterns. PMID- 11014514 TI - Salivary alterations in type 2 (non-insulin-dependent) diabetes mellitus and hypertension. AB - OBJECTIVES: The aim of this study was to determine whether saliva output and composition are altered in type 2 diabetes mellitus by comparison with a healthy, non-medicated control group, and also a group of hypertensives. METHODS: From a community-dwelling cohort of Mexican American and European American subjects enrolled in the OH:SALSA oral aging study, we identified 233 subjects with type 2 diabetes mellitus, 227 with hypertension, and 240 healthy control subjects. We collected unstimulated whole (UW) and submandibular/ sublingual (US) saliva, as well as stimulated parotid (SP) and submandibular/ sublingual (SS) saliva. Flow rates were determined, yeast carriage was assayed in UW saliva, and SP and SS saliva samples were analyzed for protein composition. ELISA was used to determine concentrations of an array of specific protein components, with both antimicrobial and other activities. RESULTS: Both diabetic and hypertensive subjects had reduced output of both stimulated and unstimulated submandibular/sublingual saliva. 30% of the diabetic subjects had high oral yeast counts (> or =1000 cfu/mL) compared with 17% of the healthy subjects and 20% of the hypertensives. Significant increases in the concentrations of a number of the protein components were found in the diabetic subjects, specifically, SP lactoferrin, myeloperoxidase (MPO), and salivary peroxidase (SPO), as well as SS total protein, albumin, lactoferrin and secretory IgA. CONCLUSIONS: The pattern of decreased flow rates and increased protein concentrations were similar, but consistently greater in diabetics than hypertensives, suggesting that disease specific mechanisms may be responsible. Diabetics may be more prone to oral dryness and infections than non-diabetics. PMID- 11014515 TI - Decline of caries prevalence after the cessation of water fluoridation in the former East Germany. AB - In contrast to the anticipated increase in dental caries following the cessation of water fluoridation in the cities Chemnitz (formerly Karl-Marx-Stadt) and Plauen, a significant fall in caries prevalence was observed. This trend corresponded to the national caries decline and appeared to be a new population wide phenomenon. Additional surveys (N=1017) carried out in the formerly fluoridated towns of Spremberg (N=9042) and Zittau (N=6232) were carried out in order to support this unexpected epidemiological finding. Pupils from these towns, aged 8/9-, 12/13- and 15/16-years, have been examined repeatedly over the last 20 years using standardised caries-methodological procedures. While the data provided additional support for the established fact of a caries reduction brought about by the fluoridation of drinking water (48% on average), it has also provided further support for the contention that caries prevalence may continue to fall after the reduction of fluoride concentration in the water supply from about 1 ppm to below 0.2 ppm F. Caries levels for the 12-year-olds of both towns significantly decreased during the years 1993-96, following the cessation of water fluoridation. In Spremberg, DMFT fell from 2.36 to 1.45 (38.5%) and in Zittau from 2.47 to 1.96 (20.6%). These findings have therefore supported the previously observed change in the caries trend of Chemnitz and Plauen. The mean of 1.81 DMFT for the 12-year-olds, computed from data of the four towns, is the lowest observed in East Germany during the past 40 years. The causes for the changed caries trend were seen on the one hand in improvements in attitudes towards oral health behaviour and, on the other hand, to the broader availability and application of preventive measures (F-salt, F-toothpastes, fissure sealants etc.). There is, however, still no definitive explanation for the current pattern and further analysis of future caries trends in the formerly fluoridated towns would therefore seem to be necessary. PMID- 11014516 TI - Oral mucosal lesions in a representative cross-sectional study of aging Germans. AB - OBJECTIVES: To determine prevalence of oral mucosal lesions in a cross-sectional study among aging Germans. METHODS: Three specially trained dental teams examined adults (35-44 yrs, Group 1) and senior individuals (65-74 yrs, Group 2) in 90 sample points of which 60 were located in the former West and 30 in the former East part of Germany. The spectrum comprised 28 different oral lesions with subforms. RESULTS: 655 individuals in Group 1 (35-44 yrs) and 1367 individuals in Group 2 (65-74 yrs) were studied. 33.8% (Group 1) and 33.9% (Group 2) were without any pathology of the oral mucosa. Several lesions were not recorded in both Groups like oral hairy leukoplakia and gingival hyperplasia (Group 1 and two) and xerostomia (Group 1). In Group 1 history for labial herpetic lesions (31.7%), Fordyce granules (26.6%), history for recurrent aphthous ulceration (18.3%) and lip and/or cheek biting (10.1%) were recorded. In Group 2 Fordyce granules (23.7%), history of labial herpes (20.0%), plicated tongue (19.0%) and denture stomatitis (18.3%) were those lesions most frequently recorded. Leukoplakia was seen in 1.8% (West) and 0.9% (East) respectively; men were more often affected than women (2.3% versus 0.0% P<0.05, Group 1; 2.3% versus 0.9%, Group 2. There was association between the prevalence of leukoplakia and a lower (3.3%) or higher educational level (0.5%). Denture associated lesions were seen in 18.3% (Group 2) compared to 2.5% (Group 1) (P<0.001). Other age-related lesions were lip and/or cheek biting being more prevalent in Group 1 10.1% versus 1.9% (P<0.001), plicated tongue 19.0% in Group 2 versus 3.8% in Group 1 (P<0.001). CONCLUSIONS: The present study has shown prevalence to be comparable to other relevant Western European studies. Since the spectrum of oral mucosal lesions changes with age and increases with general morbidity, routine examinations of oral cavities of the aging are mandatory particularly to detect early precancerous and other mucosal lesions. PMID- 11014517 TI - Multitoxin biosensor-mass spectrometry analysis: a new approach for rapid, real time, sensitive analysis of staphylococcal toxins in food. AB - Biomolecular interaction analysis mass spectrometry (BIA-MS) was applied to detection of bacterial toxins in food samples. This two-step approach utilizes surface plasmon resonance (SPR) to detect the binding of the toxin(s) to antibodies immobilized on a surface of a sensor chip. SPR detection is then followed by identification of the bound toxin(s) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Staphylococcal enterotoxin B (SEB) was readily detected in milk and mushroom samples at levels of 1 ng/ml. In addition, non-specific binding of food components to the immobilized antibody and to the sensor chip surface was detected. To evaluate the applicability of BIA-MS in the analysis of materials containing multiple toxic components, sample containing both SEB and toxic-shock syndrome toxin-1 was analyzed. Both toxins were successfully and simultaneously detected through the utilization of multiaffinity sensor chip surfaces. PMID- 11014518 TI - Microbial species associated with different sections of broccoli harvested from three regions in Australia. AB - The microbial populations associated with the different sections of broccoli harvested from three locations in Australia were studied during storage at 5, 15 and 20 degrees C. Bacterial and yeast populations associated with the outer florets and cut surfaces of the stem were generally 10-fold or more higher than those associated with inner florets or non-cut stems, respectively. The predominating bacterial species varied with the origin of the broccoli. Pseudomonas fluorescens, Ps. corrugata and Ps. viridiflava predominated at populations of 10(5)-10(7) cfu/g on broccoli harvested from Victoria, Ps. fluorescens, Ps. mendocina and Ps. fragii and Arthrobacter spp. (10(-3) 10(6) cfu/g) were prevalent on broccoli harvested from Queensland. Broccoli harvested from New South Wales exhibited a predominance of Ps. fluorescens, Arthrobacter spp. and Enterobacteragglomerans (10(3)-10(5) cfu/g). Most species grew on broccoli during storage. Similar species were found at the different sections of broccoli, although, for some species there was evidence of strain variation at the different locations and for different temperature of storage. PMID- 11014519 TI - Action of lysozyme and nisin mixtures against lactic acid bacteria. AB - Lysozyme was formulated together with nisin for usage against food spoilage lactobacilli. The mixtures demonstrated improved minimal inhibitory concentrations (MIC), compared to the parent compounds, for many of the bacteria and media tested, including high salt media in which lysozyme lost virtually all of its activity. Synergy was also observed through measurement of the kinetics of bacterial killing of L. curvatus 845, in which strain synergy had been observed in MIC assays. The combination of lysozyme and nisin caused more severe cell damage as viewed by scanning electron microscopy, and a consequent change in optical density at 600 nm, compared to the parent compounds, effects that were presumed to reflect the action of lysozyme. In addition, the combination caused more rapid permeabilization (depolarization) of the cytoplasmic membranes of Staphylococcus aureus, an effect that reflected the mechanism of action of nisin. Thus, nisin and lysozyme appear to demonstrate synergy against gram-positive bacteria because they reinforce each others mechanisms of bacterial killing. PMID- 11014520 TI - Interactions of high hydrostatic pressure, pressurization temperature and pH on death and injury of pressure-resistant and pressure-sensitive strains of foodborne pathogens. AB - The objective of this study is to determine the interactions between high hydrostatic pressure, pressurization temperature, time and pH during pressurization on death and injury of pressure-resistant and pressure-sensitive strains of four foodborne pathogens: Staphylococcus aureus 485 and 765, Listeria ,monocytogenes CA and OH2, Escherichia coli O157:H7 933 and 931, Salmonella enteritidis FDA and Salmonella typhimurium E21274. Among these strains S. aureus 485, L. monocytogenes CA, E. coli O157:H7 933 and S. enteritidis FDA were reported to be more pressure-resistant than the respective strain of the same species (Alpas et al., 1999). In general, viability loss of all pathogens was enhanced significantly as the level of pressure and temperature were increased (P < 0.05). All the strains except S. aureus 485 demonstrated more than 8 log cycle reduction when pressurized at 345 MPa at 50 degrees C for 5 min. This strain seemed to be the most pressure-resistant strain within the conditions of the study. Pressurization in the presence of either citric or lactic acid increased the viability loss by an additional 1.2-3.9 log cycles at pH 4.5 for both acids at 345 MPa. This study has indicated that high hydrostatic pressure applied in conjunction with mild heat and acidity can be an effective method for inactivating pressure-resistant and pressure-sensitive strains of four foodborne pathogens in organic acid solutions. This combination treatment indicates possible pressure pasteurization applications to liquid foods that have low pH. reserved. PMID- 11014521 TI - Phenotypic and genetic diversity of Saccharomyces contaminants isolated from lager breweries and their phylogenetic relationship with brewing yeasts. AB - A taxonomic study was carried out for isolates of Saccharomyces spp. identified as contaminants ("wild yeast") in 24 different lager breweries. With reference to the current taxonomy all isolates were found to belong to the Saccharomyces sensu stricto complex and 58% of the isolates were further identified as S. cerevisiae, 26% as S. pastorianus and 3% as S. bayanus. The remaining isolates (13%) could not be identified to the species level based on their phenotypic characteristics. However, some of these isolates were identified as S. cerevisiae by HaeIII restriction digest of PCR-amplified intergenic transcribed spacer (ITS) regions. Chromosome length polymorphism (CLP) was evident among the Saccharomyces brewing contaminants with chromosome profiles typical of Saccharomyces sensu stricto. Based upon cluster analysis of their chromosome profiles the majority of the brewing contaminants could be grouped as either S. cerevisiae or S. pastorianus/S. bayanus. Further, the technique was able to differentiate between almost all brewing contaminants and to separate them from any specific lager brewing yeast. The diversity of the Saccharomyces brewing contaminants clearly demonstrated by their CLP was further reflected by MAL genotyping. For the majority of the isolates more than two MAL loci were found with MAL1, MAL2 MAL3, MAL4 and MAL11, MAL31, MAL41 as the dominant genotypes. For all isolates MAL11 and MAL31 were found whereas MAL61 only was found for one isolate. The high number of MAL loci found in the SaccharomYces brewing contaminants indicate their adaptation to a maltose-enriched environment. PMID- 11014522 TI - Characterisation of yeast flora isolated from an artisanal Portuguese ewes' cheese. AB - The evolution of the yeast flora was studied for an artisanal semi-hard ewes' cheese made from raw milk. Mean log10 yeast counts per gram of cheese body ranged from 2.7 to 6.4, with the higher counts observed after a ripening period of 30 days. The yeast population decreased thereafter and, at the end of curing process, reached values similar to those of the beginning. A total of 344 yeasts strains were randomly isolated from the curd and cheese body during the 60 days long ripening period. Esterase activity was common to almost all isolates (98%) while proteolysis was observed in 12% of the total yeast population. The proportion of strains with positive glucose fermentation increased from 21% in the curd to 75% at the end of the ripening period. A total of 150 isolates representative of the physiological characteristics tested were examined with the API ID 32C system showing different degrees of quality of identification. Only 15% of the strains (23 isolates) were excellently identified being assigned to the species Candida zeylanoides. The most frequent species appeared to be Debaryomyces hansenii (anamorph Candida famata) and Candida intermedia. These two species amounted to 9% of the yeasts in the curd increasing to 86% at the end of the ripening period. PMID- 11014523 TI - Selective enrichment broth for the isolation of Aeromonas sp. from chicken meat. AB - Six selective agents (ampicillin, novobiocin, cephalothin, bile salts, brilliant green and ethanol) were tested during the development of a selective enrichment broth for the isolation of Aeromonas sp. from food. Cephalothin at 10 mg/l was found to be the best selective agent owing to its greater selectivity and efficiency in recovering stressed and lower cell concentrations of Aeromonas sp. Higher concentrations (15-25 mg/l) of cephalothin were inhibitory to some strains of A. sobria. Cephalothin (10 mg/l) was incorporated in buffered dextrin broth (BCDB-10) and alkaline peptone water (CAPW-10) and employed for the isolation of Aeromonas sp. from chicken meat naturally and artificially inoculated (with 10(9) cells/ml of A. hydrophila). The highest isolation rate (22%) with naturally contaminated chicken was achieved with CAPW-10 in comparison to 16% with BCDB-10 and 8% with APW. Similarly, from artificially inoculated samples, 100% isolation was accomplished with CAPW-10, against 80% with BCDB-10 and 50% with APW. PMID- 11014524 TI - Chemical, physical and enzymatic pre-treatments of probiotic lactobacilli alter their adhesion to human intestinal mucus glycoproteins. AB - Intestinal mucus glycoproteins extracted from faeces of healthy adult subjects were used as a substratum for bacterial adhesion to investigate the effects of physical, chemical and enzymatic pre-treatments of the bacteria on their adhesion. The strains studied were Lactobacillus acidophilus 1 (LCI, Nestle), L. rhamnosus strain GG (ATCC 53103), L. rhamnosus LC-705, and L. casei strain Shirota (Yakult, Yakult Ltd). Hereafter the strains are referred to as LA1, LGG, LC-705, and Shirota, respectively. Strains LA1 and LGG adhered greatly whereas the adhesion of strains LC-705 and Shirota to intestinal mucus glycoproteins was low. Adhesion of LA1 and LGG was reduced by boiling, autoclaving and by pepsin and trypsin treatments suggesting that the bacterial protein structures are essential for their adhesion. Treatment in ethanol and in propanol prior to adhesion significantly increased the adhesion of LA1 and LC-705, respectively. Adhesion of Shirota strain was not altered by any of the treatments. PMID- 11014525 TI - Acid resistance of enterohaemorrhagic and generic Escherichia coli associated with foodborne disease and meat. AB - As part of the Australia New Zealand Food Authorities (ANZFA) food standards code, salami manufacturers are required to demonstrate that their process is capable of achieving a 3-log reduction in Escherichia coli. Non-pathogenic E. coli strains with similar or greater acid resistance to enterohaemorrhagic E. coli (EHEC) are needed if industry is to conduct challenge studies to demonstrate compliance with the standard. In the present study, E. coli isolates from sheep and beef carcasses and meat were shown to have wide-ranging acid resistance in broth when preadapted to growth in acidic conditions. Times required for a 3-log reduction in E. coli ranged from less than I day to more than 28 days. Variable acid resistance was observed in both EHEC strains associated with foodborne outbreaks and generic E. coli strains. Generic E. coli strains with the greatest acid resistance were assessed for pathogenicity markers and their survival in fermented meat compared with EHEC strains. It was demonstrated that generic E. coli strains could be used for challenge studies to determine compliance with or validate performance standards designed for the control of EHEC. PMID- 11014526 TI - Cluster analysis, richness and biodiversity indexes derived from denaturing gradient gel electrophoresis fingerprints of bacterial communities demonstrate that traditional maize fermentations are driven by the transformation process. AB - The bacterial communities of maize fermented foods (pozol, poto-poto and ogi) from Mexico, Congo and Benin was compared using a culture-independent approach [denaturing gradient gel electrophoresis (DGGE) analysis of total DNA]. Foods produced following the same flow chart (i) grouped in distinct clusters, (ii) shared similar richness and biodiversity indexes and (iii) exhibited a high intra specific variability. Structural biodiversity was higher in pozol samples, probably due to oxic conditions and higher initial pH. DGGE bands found in foods of different origins suggest that Lactobacillus plantarum, Lb. delbrueckii and Lb. fermentum are particularly well adapted to the fermentation of maize. PMID- 11014527 TI - Phenotypic adaptation to freeze-thaw stress of the yeast-like fungus Geotrichum candidum. AB - The effect of cold stress on Geotrichum candidum was investigated at chill and freezing temperatures. Specific growth rates were determined at various temperatures and plotted according to the Ratkowsky and Arrhenius equations. The obtained profiles led to the determination of characteristics including the activation energy and notional minimum temperatures. At temperatures below the optimum single linear slopes were observed. At freezing temperatures, the loss of viability of cell populations was proportional to the number of freezing-thawing cycles. Nevertheless, the ability of G. candidum to survive this challenge depended on the physiological conditions prior to the freezing stress. The loss of viability was growth phase specific. Cells harvested in stationary phase showed a higher resistance compared to those obtained with cells in exponential phase. Furthermore, the cells of G. candidum could be adapted to the freeze-thaw challenge by pre-treatment at chill temperatures. This phenomenon known as cryotolerance was a function of the duration of the preincubation exposure. PMID- 11014528 TI - Detection of freeze-injured Escherichia coli O157:H7 cells from foods by resuscitation prior to selective enrichment. AB - We tried to detect Escherichia coli O157:H7 in food samples artificially contaminated with freeze-injured E. coli O157:H7 using an enrichment method with modified EC broth supplemented with novobiocin (mEC + n). When the samples were cultured for enrichment immediately after inoculation of freeze-injured cells, E. coli O157:H7 was not detected in 13 out of 18 samples. However, allowing the food samples to stand for 3 h at room temperature prior to enrichment in mEC + n remarkably improved recovery of E. coli O157:H7 except for some acidic foods. E. coli O157:H7 was detected in the acidic foods by introducing a resuscitation step of 3-h of incubation in a non-selective broth at room temperature prior to enrichment with mEC + n. PMID- 11014529 TI - Do women fare worse: a metaanalysis of gender differences in traumatic brain injury outcome. AB - OBJECT: The purpose of this metaanalysis was to investigate possible gender differences in traumatic brain injury (TBI) sequelae. The case fatality rates in patients after TBI have previously been shown to be significantly higher in women as compared with men. METHODS: A quantitative review of published studies of TBI outcome revealed eight studies (20 outcome variables) of TBI, in which outcome was reported separately for men and women. CONCLUSIONS: Outcome was worse in women than in men for 85% of the measured variables, with an average effect size of -0.15. Although clinical opinion is often that women tend to experience better outcomes than men after TBI, the opposite pattern was suggested in the results of this metaanalysis. However, this conclusion is limited by the fact that, in only a small percentage of the total published reports on TBI outcome, was outcome described separately for each sex. A careful, prospective study of sex differences in TBI outcome is clearly needed. PMID- 11014530 TI - Effect of long-term mild hypothermia therapy in patients with severe traumatic brain injury: 1-year follow-up review of 87 cases. AB - OBJECT: The goal of this study was to investigate the protective effects of long term (3-14 days) mild hypothermia therapy (33-35 degrees C) on outcome in 87 patients with severe traumatic brain injury (TBI) (Glasgow Coma Scale score < or = 8). METHODS: In 43 patients assigned to a mild hypothermia group, body temperatures were cooled to 33 to 35 degrees C a mean of 15 hours after injury and kept at 33 to 35 degrees C for 3 to 14 days. Rewarming commenced when the individual patient's intracranial pressure (ICP) returned to the normal level. Body temperatures in 44 patients assigned to a normothermia group were maintained at 37 to 38 degrees C. Each patient's outcome was evaluated 1 year later by using the Glasgow Outcome Scale. One year after TBI, the mortality rate was 25.58% (11 of 43 patients) and the rate of favorable outcome (good recovery or moderate disability) was 46.51% (20 of 43 patients) in the mild hypothermia group. In the normothermia group, the mortality rate was 45.45% (20 of 44 patients) and the rate of favorable outcome was 27.27% (12 of 44 patients) (p < 0.05). Induced mild hypothermia also markedly reduced ICP (p < 0.01) and inhibited hyperglycemia (p < 0.05). The rates of complication were not significantly different between the two groups. CONCLUSIONS: The data produced by this study demonstrate that long-term mild hypothermia therapy significantly improves outcomes in patients with severe TBI. PMID- 11014531 TI - Risk of rupture from incidental cerebral aneurysms. AB - OBJECT: Controversy still exists about the risk estimation for rupture of untreated saccular aneurysms presenting for causes other than subarachnoid hemorrhage (SAH). The object of this study was to address this issue. METHODS: Between January 1976 and December 1997 in the Aizu Chuou Hospital, 62 patients underwent observation for more than 6 months for saccular, nonthrombotic, noncalcified unruptured aneurysms at locations not related to the cavernous sinus, which were detected in cerebral angiography studies performed for causes other than SAH. Clinical follow-up data in those 62 patients were reviewed to identify the risk of SAH. All patients were followed until July 1998, with the observation period ranging from 6 months to 17 years (mean 4.3 years). Seven patients (11.3%) developed SAH confirmed on computerized tomography (CT) scanning at a mean interval of 4.8 years, six of whom died and one of whom recovered with a major deficit. In addition, one patient died of the mass effect of the aneurysm, and another after sudden onset of headache and vomiting. The 5- and 10 year cumulative risks of CT-confirmed SAH calculated by the Kaplan-Meier method were 7.5% and 22.1%, respectively, for total cases, 33.5% and 55.9%, respectively, for large (> 10 mm) aneurysms, and 4.5% and 13.9%, respectively, for small (< 10 mm) aneurysms. CONCLUSIONS: Although based on a relatively small, single-institution series, our data indicated that the risk of rupture from incidental, intradural, saccular aneurysms was higher than previously reported, and may support preventive surgical treatment of incidental aneurysms, considering the fatality rate of SAH. PMID- 11014532 TI - Intraoperative endovascular treatment as an adjunct to microsurgical clipping of paraclinoid aneurysms. AB - OBJECT: The endovascular procedure can provide proximal control, suction decompression, and prompt intraoperative angiography during microsurgical clipping of aneurysms of the paraclinoid segment of the internal carotid artery (ICA). The authors assess the safety and feasibility of this method in 24 consecutive cases. METHODS: Frontotemporal craniotomy and radical pterionectomy were performed with the patient's head immobilized in a radiolucent frame while femoral artery catheterization was achieved. Before dural opening, a balloon catheter with a coaxial lumen was positioned and tested in the ICA, after which microsurgical exposure was completed, including intradural clinoid drilling and optic canal decompression. Trapping of the lesion was achieved by inflating the balloon and placing a temporary clip beyond the aneurysm neck. The catheter was gently aspirated to achieve suction decompression and to facilitate clip application. Intraoperative digital subtraction angiography was then performed. Twenty-two aneurysms were larger than 10 mm, and 11 of them were giant. Six patients presented with subarachnoid hemorrhage and nine with visual symptoms. Balloon occlusion and suction decompression were performed in 16 cases (67%), and proximal control alone in 1 case. Intraoperative angiography was performed in every case. Subsequent clip readjustment was necessary in seven cases, including three cases of residual aneurysm filling and four of ICA compromise. Complete aneurysm obliteration was achieved in 20 cases, and greater than 90% obliteration in 22. One major infarct likely related to catheter thromboembolism was found. There were no instances of visual deterioration or other complications attributable to the endovascular procedure. CONCLUSIONS: The endovascular method allows safe and reliable proximal control, suction decompression, and intraoperative angiography in microsurgical treatment of large paraclinoid aneurysms. PMID- 11014533 TI - Natural history of the neck remnant of a cerebral aneurysm treated with the Guglielmi detachable coil system. AB - OBJECT: The long-term durability of Guglielmi detachable coil (GDC) embolization of cerebral aneurysms is still unknown. The purpose of this study was to evaluate the anatomical evolution of neck remnants in aneurysms treated with GDCs. METHODS: Of 455 aneurysms treated with GDCs from 1990 to 1998 at the University of California at Los Angeles Medical Center, 178 aneurysms (39%) had residual necks postembolization. Long-term follow-up angiograms were obtained in 73 of these aneurysms in 71 patients. The mean duration of angiographic follow up was 17.3 months. Twenty-four of the aneurysms were small with small necks, 24 were small with wide necks, 15 were large, and 10 were giant aneurysms. In small aneurysms with small necks, postembolization angiography revealed 12 aneurysms (50%) with progressive thrombosis, eight (33%) unchanged, and four (17%) with recanalization. In small aneurysms with wide necks, six (25%) had progressive thrombosis, eight (33%) remained unchanged, and 10 (42%) had recanalization. In large aneurysms, two (13%) were unchanged and 13 (87%) had recanalization. Of the giant aneurysms only one (10%) remained unchanged and nine (90%) had recanalization. Overall, 18 aneurysms (25%) exhibited progressive thrombosis, 19 (26%) remained unchanged, and 36 (49%) displayed recanalization on follow-up angiography. During the last 2 years of the study, the recanalization rate decreased and a higher rate of progressive thrombosis was noted in aneurysms with small necks. These positive changes are related to important new technical developments. CONCLUSIONS: Treatment with GDCs appears to be effective and the results permanent in most small aneurysms with small necks. However, there are important technical limitations in the current GDC technology that prevent recanalization in wide-necked or large or giant aneurysms. PMID- 11014534 TI - Impact of Guglielmi detachable coils on outcomes of patients with intracranial aneurysms treated by a multidisciplinary team at a single institution. AB - OBJECT: The goal of this study was to investigate the impact of the introduction of the Guglielmi detachable coil (GDC) therapeutic option on the overall management outcome of intracranial aneurysms. The authors accomplished this by assessing patient morbidity and mortality, inflation-adjusted hospital charges, lengths of stay in the hospital and the intensive care unit (ICU), and treatment efficacy. METHODS: The authors conducted a retrospective analysis of consecutive cases of intracranial intradural aneurysms managed by a single multidisciplinary neurovascular team at a tertiary care, academic referral center during the 24 months preceding the introduction of the GDC procedure (Group I or pre-GDC era, 77 patients) and during the first 24 months after its introduction (Group II or GDC era, 99 patients). Treatment with GDCs was considered for cases of higher clinical grade or poor surgical risk, or in response to patient preference (27 [27%] of 99 patients in Group II). Host and lesion parameters in our cohort were validated against outcome parameters by using univariate and multivariate analyses. The pre-GDC and GDC subgroups of patients were comparable for major disease severity parameters (patient age, lesion location, clinical grade, and hemorrhage severity). There was no significant difference in clinical outcome at 6 months, infarcts on computerized tomography scanning, or aneurysm obliteration rates before and after introduction of GDC treatment. Decreasing trends in duration of hospital and ICU stay and in inflation-adjusted hospital charges occurred well before and thus were unrelated to the introduction of the GDC therapeutic option. CONCLUSIONS: The results of this study do not demonstrate any significant impact of integration of the GDC modality on clinical outcome, mortality, morbidity, or effectiveness of treatment. Ongoing improvements in hospital charges and length of hospital stay appeared unrelated to the introduction of the GDC option. PMID- 11014535 TI - Change in circulating blood volume following craniotomy. AB - OBJECT: The importance of monitoring circulating blood volume (CBV) during perioperative management is widely recognized in critically ill patients. The purpose of this study was to investigate the change in CBV following craniotomy by using indocyanine-green pulse spectrophotometry. METHODS: Circulating blood volume and plasma hormones related to stress and fluid regulation were measured five times: preoperatively, immediately postoperatively, and 1, 2, and 7 days after craniotomy was performed in 17 patients with a brain tumor or an unruptured aneurysm. The mean value of CBV preoperatively was 82 ml/kg, which decreased to 64 ml/kg (78%) immediately postoperatively and gradually recovered to 82 ml/kg on Day 7 postsurgery (p = 0.0069). The mean values of adrenaline, noradrenaline, arginine vasopressin, renin, and aldosterone were highest immediately postoperatively. The mean intraoperative balances of water and sodium were 1,090 ml and 113 mEq, respectively. Partial correlation coefficients of CBV to noradrenaline and serum sodium during the entire study were -0.430 (p = 0.0036) and 0.418 (p = 0.0048), respectively. CONCLUSIONS: Attention should be paid to decreased CBV following craniotomy, which is caused by the shift of fluid to interstitial spaces due to surgical stress. Hypovolemia can be suspected from a postoperative decrease in serum sodium. PMID- 11014536 TI - Continuous electromyography monitoring of motor cranial nerves during cerebellopontine angle surgery. AB - OBJECT: Electromyography (EMG) monitoring is expected to reduce the incidence of motor cranial nerve deficits in cerebellopontine angle surgery. The aim of this study was to provide a detailed analysis of intraoperative EMG phenomena with respect to their surgical significance. METHODS: Using a system that continuously records facial and lower cranial nerve EMG signals during the entire operative procedure, the authors examined 30 patients undergoing surgery on acoustic neuroma (24 patients) or meningioma (six patients). Free-running EMG signals were recorded from muscles targeted by the facial, trigeminal, and lower cranial nerves, and were analyzed off-line with respect to waveform characteristics, frequencies, and amplitudes. Intraoperative measurements were correlated with typical surgical maneuvers and postoperative outcomes. Characteristic EMG discharges were obtained: spikes and bursts were recorded immediately following the direct manipulation of a dissecting instrument near the cranial nerve, but also during periods when the nerve had not yet been exposed. Bursts could be precisely attributed to contact activity. Three distinct types of trains were identified: A, B, and C trains. Whereas B and C trains are irrelevant with respect to postoperative outcome, the A train--a sinusoidal, symmetrical sequence of high-frequency and low-amplitude signals--was observed in 19 patients and could be well correlated with additional postoperative facial nerve paresis (in 18 patients). CONCLUSIONS: It could be demonstrated that the occurrence of A trains is a highly reliable predictor for postoperative facial palsy. Although some degree of functional worsening is to be expected postoperatively, there is a good chance of avoiding major deficits by warning the surgeon early. Continuous EMG monitoring is superior to electrical nerve stimulation or acoustic loudspeaker monitoring alone. The detailed analysis of EMG-waveform characteristics is able to provide more accurate warning criteria during surgery. PMID- 11014537 TI - Role of hydrodynamic processes in the pathogenesis of peritumoral brain edema in meningiomas. AB - OBJECT: In a prospective study, 28 patients with 32 intracranial meningiomas were examined to determine the role of hydrodynamic interaction between tumor and surrounding brain tissue in the pathogenesis of peritumoral brain edema. METHODS: Gadolinium-diethylenetriamine pentaacetic acid (Gd-DPTA), an extracellular contrast agent used for routine clinical imaging, remains strictly extracellular without crossing an intact blood-brain barrier. Therefore, it is well suited for investigations of hydrodynamic extracellular mechanisms in the development of brain edema. Spin-echo T1-weighted magnetic resonance images were acquired before and after intravenous administration of 0.2 mmol/kg Gd-DPTA. Additional T1 weighted imaging was performed 0.6, 3.5, and 6.5 hours later. No significant Gd DPTA diffused from tumor into peritumoral brain tissue in 12 meningiomas without surrounding brain edema. In contrast, in 17 of 20 meningiomas with surrounding edema, contrast agent in peritumoral brain tissue was detectable after 3.5 hours and 6.5 hours. In three of 20 meningiomas with minimum surrounding edema (<5 cm3), contrast agent effusion was absent. After 3.5 hours and 6.5 hours strong correlations of edema volume and the maximum distance of contrast spread from the tumor margin into adjacent brain parenchyma (r = 0.84 and r = 0.87, respectively, p < 0.0001) indicated faster effusion in larger areas of edema. CONCLUSIONS: The results of this study show that significant contrast agent effusion from the extracellular space of the tumor into the interstitium of the peritumoral brain tissue is only found in meningiomas with surrounding edema. This supports the hypothesis that hydrodynamic processes play an essential role in the pathogenesis of peritumoral brain edema in meningiomas. PMID- 11014539 TI - Posture-related changes in the pressure environment of the ventriculoperitoneal shunt system. AB - OBJECT: The purpose of this study is to clarify the whole pressure environment of the ventriculoperitoneal (VP) shunt system in patients with successfully treated hydrocephalus and to determine which factor of the pressure environment has a preventive effect on overdrainage. METHODS: Thirteen patients with hydrocephalus who had been treated with VP shunt therapy by using a Codman-Hakim programmable valve without incidence of overdrainage were examined. The authors evaluated intracranial pressure (ICP), intraabdominal pressure (IAP), hydrostatic pressure (HP), and the perfusion pressure (PP) of the shunt system with the patients both supine and sitting. With patients supine, ICP, IAP, and HP were 4.6 +/- 3 mm Hg, 5.7 +/- 3.3 mm Hg, and 3.3 +/- 1 mm Hg, respectively. As a result, the PP was only 2.2 +/- 4.9 mm Hg. When the patients sat up, the IAP increased to 14.7 +/- 4.8 mm Hg, and ICP decreased to-- 14.2 +/- 4.5 mm Hg. The increased IAP and decreased ICP offset 67% of the HP (42.9 +/- 3.5 mm Hg), and consequently the PP (14 +/- 6.3 mm Hg) corresponded to only 33% of HP. CONCLUSIONS: The results observed in patients indicated that IAP as well as ICP play an important role in VP shunt therapy and that the increased IAP and the decreased ICP in patients placed in the upright position allow them to adapt to the siphoning effect and for overdrainage thereby to be avoided. PMID- 11014538 TI - Prognostic factors in intracranial ependymomas in children. AB - OBJECT: The occurrence of intracranial ependymomas in children is relatively infrequent, and their prognostic factors are still controversial, especially regarding histological composition. METHODS: A retrospective study was conducted of 37 children treated during the last 20 years for intracranial ependymomas at the Hopital de la Timone. Both univariate and multivariate statistical analyses were performed to assess the prognostic relevance of patient age and sex, extent of tumor removal, location of the tumor (supratentorial compared with infratentorial, median compared with lateral), tumor histological composition, and adjuvant therapies in affecting the 5-year progression-free survival (PFS) rate and overall survival (OS) rate. The following histopathological features, either alone or in combination, were analyzed: endothelial proliferation, necrosis, loss of differentiating structures (present compared with absent), the number of mitotic figures per 10 hpf, and cellularity (number of nuclei/5 hpf). In addition, immunohistochemical detection of Ki-67 antigen was performed and the Ki-67 labeling index (LI) evaluated in all cases. The 5-year OS and PFS rates were 45% and 25%, respectively (median follow up 34 months). Four patients died of disease without remission (median 163 days) and disease in 21 patients relapsed: 18 in situ and three both in situ and distantly. On univariate analysis total surgical resection and median infratentorial location were associated with a better outcome (p < 0.002) for both OS and PFS. Loss of differentiating structures was associated with poor prognosis (p < 0.008) and the combination of necrosis, endothelial proliferation, and mitotic index greater than 5 was also a negative predictive factor for both OS (p < 0.002) and PFS (p = 0.02). The PFS time was shorter in patients younger than 4 years of age and in patients in whom a Ki-67 LI greater than 1 was found (p = 0.03 and 0.006, respectively). Adjuvant radiotherapy and chemotherapy were not relevant to prognosis. Moreover, among the 15 patients in whom total excision was achieved, OS was better in those who did not receive adjuvant therapies. In contrast, adjuvant therapies significantly enhanced PFS time in patients in whom tumor excision was incomplete. CONCLUSIONS: This study and analysis of the literature further highlight that total tumor removal is the treatment of choice for ependymomas in children. Postoperative measurement of residual tumor is required, especially because a subgroup of patients might be treated by surgery alone. Median infratentorial ependymomas have to be distinguished from the lateral type. Appropriate and reproducible histological parameters and Ki-67 LI are of interest as predictors of outcome. PMID- 11014540 TI - Telomerase activity in primary and secondary glioblastomas multiforme as a novel molecular tumor marker. AB - OBJECT: Telomerase activity is responsible for cell immortality. To examine the role of telomerase in the carcinogenesis of human glioblastomas multiforme (GBMs), the authors studied telomerase activity, telomerase component expression, and telomere lengths in 42 GBM samples. METHODS: In all samples, EGFR and MDM2 amplifications and overexpressions were examined using Southern and Northern blot analyses. The p53 mutation was analyzed using polymerase chain reaction-single strand conformational polymorphism and by direct sequence analysis. Specimens of tissues were immunostained with p53, EGFR, and MDM2 antibodies. Allelic loss on chromosomes 17p and 10 was assessed by loss of heterozygosity (LOH) assays. Telomerase activity, expression of its components (human telomerase reverse transcriptase [hTERT], human telomerase RNA component [hTERC], and telomerase associated protein [TEP1]), and telomere lengths were analyzed using the telomeric repeat amplification protocol (TRAP)-hybridization protection assay, reverse transcription-polymerase chain reaction, and Southern blot analysis. According to the results of assessments of EGFR and MDM2 amplifications, p53 mutation, LOHs in chromosomes 17p and 10, and the clinical course of the disease, the 42 samples were classified into 22 primary and 20 secondary glioblastomas. Twenty-six (61.9%) of all 42 samples demonstrated detectable telomerase activity during the TRAP assay. Secondary GBMs displayed significantly higher levels of telomerase activity and hTERT expression than primary GBMs. Tumors with a p53 gene mutation demonstrated significantly higher telomerase activity than those without a p53 mutation. Four samples with a codon 175 mutation demonstrated an exceptionally high amount of telomerase activity. In secondary GBMs, the increase in telomerase activity and the hTERT expression level correlated with the increased frequency of p53 mutations. There was no significant difference in telomere length between primary and secondary GBMs. CONCLUSIONS: These results suggest that telomerase activity and p53 mutations both play important roles in the multistep carcinogenesis of GBMs. Telomerase activity and hTERT expression may be considered as novel distinctive factors in human GBMs. PMID- 11014541 TI - Diffuse axonal injury due to lateral head rotation in a rat model. AB - OBJECT: The authors investigated the ramifications of producing diffuse axonal injury (DAI) by lateral head rotation in a rat model. METHODS: Using a special injury-producing device, the rat's head was rapidly rotated 90 degrees in the coronal plane at an angular velocity of at least 753.13 rad/second and an angular acceleration of at least 1.806 x 10(5) rad/second2; the rotation was complete within 2.09 msec. There were no statistically significant changes in PO2, PCO2, pH, or blood pressure values at 5, 15, or 60 minutes after head rotation compared with their respective preinjury baseline values. The rats exhibited posttraumatic behavior suppression for an average of 12.6 minutes. The mortality rate was 17%. The rats that survived had diffuse subarachnoid hemorrhage around the brainstem and upper cervical cord, but no obvious brain contusion. In sections stained with silver or hematoxylin and eosin, axonal swelling and bulblike protrusions at the axonal axis were observed in the medulla oblongata, midbrain, upper cervical cord, and corpus callosum between 6 hours and 144 hours postinjury. The axonal injuries were most severe in the brainstem and were accompanied by parenchymal bleeding. The density of bulblike axonal protrusions peaked 6 hours postinjury in the medulla oblongata and 24 hours postinjury in the midbrain. CONCLUSIONS: Rapid lateral head rotation can produce DAI characterized by severe damage to the rat brainstem. PMID- 11014542 TI - Dexamethasone-induced abolition of the inflammatory response in an experimental glioma model: a flow cytometry study. AB - OBJECT: Commonly used for management of cerebral edema in patients with brain tumors, steroid medications also have immunosuppressive functions. To characterize the effects of steroids on the central nervous system's response to tumors more clearly, flow cytometry was used to quantify the extent of inflammatory cell infiltration in an immunogenic rat glioma model. METHODS: Freshly prepared 11-day-old intracranial C6 tumors that had been excised from dexamethasone-treated and untreated rats were labeled ex vivo with monoclonal antibodies against CD 11b/c, CD45, and CD8a antigens. The extent of microglia (CD11b/c-highly positive, CD45-slightly positive cell), macrophage (CD11b/c highly positive, CD45-highly positive cell), lymphocyte (CD11b/c-negative, CD45 highly positive cell), and cytotoxic T-cell (CD8a-positive cell) infiltration into each rat's tumor, tumor periphery, and contralateral tumor-free hemisphere was analyzed using flow cytometry. Microglia and lymphocytes constituted a significant component of infiltrating cells in this model, comprising 23 +/- 3% and 33 +/- 5% of viable cells, respectively. Macrophages, on the other hand, accounted for only 9 +/- 1% of infiltrating cells. Treatment of rats with a 7-day course of low-dose dexamethasone (0.1 mg/kg/day) resulted in a greater than 50% inhibition of microglia (p = 0.03) and lymphocyte (p = 0.001) infiltration into tumors. Increasing the dexamethasone dose to 1 mg/kg/day further abolished lymphocyte infiltration (89% inhibition, p = 0.001) but had no additional inhibitory effect on microglia invasion. Macrophage infiltration of tumors was not inhibited at the dexamethasone doses used in this study (p = 0.42). CONCLUSIONS: Flow cytometry is a valuable technique for characterizing tumor associated inflammatory cells in gliomas. Even at low doses, dexamethasone was found to inhibit significantly the infiltration of brain tumors by lymphocytes and microglia. These findings should be considered when experimental immunotherapeutic strategies are evaluated for clinical application. PMID- 11014543 TI - Role of calcium channels in oxyhemoglobin-induced apoptosis in endothelial cells. AB - OBJECT: Oxyhemoglobin (OxyHb) released from hemolysed erythrocytes has been considered to be responsible for cerebral vasospasm after subarachnoid hemorrhage. The authors previously reported that OxyHb produced apoptosis in cultured vascular endothelial cells. The change in intracellular Ca++ homeostasis was expected to be one of the possible mechanisms of the cytotoxic effects of OxyHb. This study was undertaken to investigate the protective effects of Ca++ channel blockers on OxyHb-induced apoptosis. METHODS: Cultured bovine coronary artery and brain microvascular endothelial cells (passages 5-9) were used. A cell density study, immunohistochemical staining, and DNA fragmentation analysis were performed to confirm apoptosis. Various concentrations (1-50 microM) of OxyHb were used for 24- to 72-hour incubations with and without Ca++-channel blockers. Oxyhemoglobin produced cytotoxicity leading to cell detachment from the culture dish in time- and concentration-dependent manners. The highest dose (50 microM) of OxyHb produced cell detachment after a 24-hour incubation, and the lower doses (1-10 microM) produced cell detachment after 48 to 72 hours. Immunohistochemical analysis showed that apoptosis occurred in cells that were still attached to the side of the culture dish after 48 to 72 hours of OxyHb treatment (5 microM). The OxyHb (10 microM) produced DNA ladders at 48 to 72 hours. Three Ca++-channel blockers were used to prevent the toxic effect of OxyHb. The voltage-dependent Ca++-channel blocker nicardipine (1 microM), the voltage-independent Ca++-channel blocker econazole (10 microM), and the inorganic Ca++-channel blocker lanthanum (100 microM) all failed to prevent cell detachment or DNA ladders produced by OxyHb. These results were similar in both cell lines. CONCLUSIONS: Oxyhemoglobin produced apoptotic changes in cultured vascular endothelial cells, and Ca++ channel blockers did not prevent OxyHb-induced apoptosis. PMID- 11014544 TI - Relationship between residual cerebral blood flow and oxygen metabolism as predictive of ischemic tissue viability: sequential multitracer positron emission tomography scanning of middle cerebral artery occlusion during the critical first 6 hours after stroke in pigs. AB - OBJECT: The authors tested the hypothesis that oxygen metabolism is the key factor linking the long-term viability of ischemic brain tissue to the magnitude of residual blood flow during the first 6 hours following a stroke. METHODS: Eleven anesthetized pigs underwent a series of positron emission tomography studies to measure cerebral blood flow (CBF) and metabolism before and for 7 hours after the animals were subjected to permanent middle cerebral artery (MCA) occlusion. The extent of collateral blood supply was assessed using angiography. Abnormal metabolism of the ischemic tissue progressed as a function of time in inverse proportion to the magnitude of residual CBF, and the volume of the infarct grew in inverse proportion to the residual blood supply. Ten hours after occlusion of the MCA, the infarct topographically matched the tissue with a cerebral metabolic rate of oxygen consumption below 50% of values measured on the contralateral side. This was also the threshold for the decline of the oxygen extraction fraction below normal, which was critical for the prediction of nonviable ischemic tissue. Mildly ischemic tissue (CBF > 30 ml/100 g/min) did not reach the cerebral metabolic rate of oxygen threshold of viability during the first 6 hours after MCA occlusion; moderately ischemic tissue (CBF 12-30 m1/100 g/ min) reached the threshold of viability in 3 hours; and severely ischemic tissue (CBF < 12 ml/100 g/min) remained viable for less than 1 hour. CONCLUSIONS: The relationship between the residual CBF and both oxygen metabolism and extraction is critical to the evolution of metabolic deficiency and lesion size after stroke. PMID- 11014545 TI - Products of hemolysis in the subarachnoid space inducing spreading ischemia in the cortex and focal necrosis in rats: a model for delayed ischemic neurological deficits after subarachnoid hemorrhage? AB - OBJECT: The pathogenesis of delayed ischemic neurological deficits after subarachnoid hemorrhage has been related to products of hemolysis. Topical brain superfusion of artificial cerebrospinal fluid (ACSF) containing the hemolysis products K+ and hemoglobin (Hb) was previously shown to induce ischemia in rats. Superimposed on a slow vasospastic reaction, the ischemic events represent spreading depolarizations of the neuronal-glial network that trigger acute vasoconstriction. The purpose of the present study was to investigate whether such spreading ischemias in the cortex lead to brain damage. METHODS: A cranial window was implanted in 31 rats. Cerebral blood flow (CBF) was measured using laser Doppler flowmetry, and direct current (DC) potentials were recorded. The ACSF was superfused topically over the brain. Rats were assigned to five groups representing different ACSF compositions. Analyses included classic histochemical and immunohistochemical studies (glial fibrillary acidic protein and ionized calcium binding adaptor molecule) as well as a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay. Superfusion of ACSF containing Hb combined with either a high concentration of K+ (35 mmol/L, 16 animals) or a low concentration of glucose (0.8 mmol/L, four animals) reduced CBF gradually. Spreading ischemia in the cortex appeared when CBF reached 40 to 70% compared with baseline (which was deemed 100%). This spreading ischemia was characterized by a sharp negative shift in DC, which preceded a steep CBF decrease that was followed by a slow recovery (average duration 60 minutes). In 12 of the surviving 14 animals widespread cortical infarction was observed at the site of the cranial window and neighboring areas in contrast to findings in the three control groups (11 animals). CONCLUSIONS: The authors conclude that subarachnoid Hb combined with either a high K+ or a low glucose concentration leads to widespread necrosis of the cortex. PMID- 11014546 TI - Microanatomy of the pericallosal arterial complex. AB - OBJECT: The pericallosal arterial complex supplies the callosal and pericallosal regions, as well as the anterior two thirds of the medial and superomedial aspects of both hemispheres. It is composed of the pericallosal artery (that is, the segment of the anterior cerebral artery located distal to the anterior communicating artery [ACoA]) and the median callosal artery (or third pericallosal artery), which originates from the ACoA. This system was studied in 46 specimens (23 human cadaver heads) injected with colored latex. METHODS: After being injected with colored latex, embalmed, and bleached, the specimens were studied with the aid of optic magnification. The pericallosal artery was found to be divided into four segments (A2-A5 in the proximodistal direction). After giving rise to central, callosal, and cortical branches, it terminated near the splenium of the corpus callosum as the posterior pericallosal artery, or on the precuneus as the inferomedial parietal artery. CONCLUSIONS: The authors propose a logical classification of the different variations in the pericallosal arterial complex based on embryological development. This complex can be considered a hemodynamic solution to an abnormal regression of one of its parts, which is balanced by the development of supplemental channels from other parts. PMID- 11014547 TI - Optical imaging of bilingual cortical representations. Case report. AB - The organization of language in the brains of multilingual persons remains controversial. The authors investigated language representations in a proficient bilingual patient by using a novel neuroimaging technique, intraoperative optical imaging of intrinsic signals (iOIS), and a visual object naming task. The results indicate that there are cortical areas that are activated by the use of both English and Spanish languages (superior temporal sulcus, superior and middle temporal gyri, and parts of the supramarginal gyrus). In addition, language specific areas were identified in the supramarginal (Spanish) and precentral (English) gyri. These results suggest that cortical language representations in bilingual persons may consist of both overlapping and distinct components. Furthermore, this study demonstrates the utility of iOIS in detecting topographical segregation of cognitively distinct cortices. PMID- 11014548 TI - Ependymal cyst producing alpha-fetoprotein. Case report. AB - This 17-year-old man was admitted to the hospital due to progressive headache and diplopia. Neuroradiological studies revealed a cystic mass in the pineal region without a parenchymal lesion. In addition, serum alpha-fetoprotein (AFP) levels were elevated. A cyst-to-third-ventricle and cistern fenestration was performed, but the cyst enlarged 3 months after the first operation. In the second operation, subtotal resection of the cyst was performed. The AFP level in the cyst fluid was very high preoperatively but was decreased postoperatively. The patient was discharged with no neurological deficit. Pathological examination of resected tissue showed a single layer of cuboidal cells that resembled an ependymal structure. The cells were immunoreactive for AFP immunostain, which indicated AFP production from these cells. PMID- 11014549 TI - Middle meningeal artery embolization for refractory chronic subdural hematoma. Case report. AB - The authors present a case of refractory chronic subdural hematoma (CSH) in a 59 year-old man with coagulopathy due to liver cirrhosis. The patient was successfully treated by embolization of the middle meningeal artery after several drainage procedures. This new therapeutic approach to recurrent CSH is discussed. PMID- 11014550 TI - Treatment of spontaneous carotid-cavernous fistula in Ehlers-Danlos syndrome by transvenous occlusion with Guglielmi detachable coils. Case report and review of the literature. AB - The authors report on a young woman with Ehlers-Danlos syndrome (EDS) Type IV in whom a spontaneous direct carotid-cavernous fistula (CCF) was treated by transvenous occlusion with regular and fiber-coated Guglielmi detachable coils. To the authors' knowledge, this is the first time this approach has been used in a patient with EDS. The different treatment options are discussed, and the literature on endovascular treatment of direct CCFs in EDS is reviewed. PMID- 11014551 TI - Synovitis, acne, pustulosis, hyperostosis, and osteitis syndrome presenting as a primary calvarial lesion. Case report and review of the literature. AB - The synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome is a recently described, currently evolving clinical entity that groups together several idiopathic disorders of bone and skin formerly described under a variety of names. Among the spectrum of possible locations for the bone lesions, there is no previous report in the literature of primary involvement of the skull vault. A patient with primary involvement of the calvaria in the setting of SAPHO syndrome is described here, which, to the authors' knowledge, is the first report of such localization. The clinically and radiologically benign evolution of the different stages of the bone lesions is presented. The authors suggest that the SAPHO syndrome should be considered in the differential diagnosis of lytic, sclerotic, or hyperostotic lesions of the skull, particularly before considering invasive diagnostic procedures. PMID- 11014552 TI - Neuroblastomatous recurrence of ganglioglioma. Case report. AB - This case is believed to be the first reported recurrent intracranial ganglioglioma with purely neuroblastomatous malignant transformation. A complete macroscopic resection of a right frontal lobe tumor in an 18-year-old woman revealed differentiated ganglioglioma. Seven years later a large, well-demarcated recurrent tumor was again macroscopically totally resected in the same patient. Histological analysis showed malignant transformation in only the neuronal component of the original tumor. A review of the literature on recurrent gangliogliomas and their malignant transformation is included. PMID- 11014553 TI - Multicentric intracranial smooth-muscle tumor in a woman with human immunodeficiency virus. Case report. AB - A 31-year-old woman with acquired immunodeficiency syndrome (AIDS) was found to harbor both a pulmonary smooth-muscle tumor and an intracranial extraaxial smooth muscle tumor of the parasellar region. The frequency of smooth-muscle tumors (leiomyoma and leiomyosarcoma) has increased with AIDS, but much more so in children than in adults. Only nine cases of human immunodeficiency virus-related smooth-muscle tumors have been previously reported in adults, and only one of these was located intracranially. PMID- 11014554 TI - Electrophysiological localization of the substantia nigra in the parkinsonian nonhuman primate. AB - During ablative surgery and implantation of deep-brain stimulators for the treatment of movement disorders, electrophysiological techniques are often used for localization of subcortical targets. New restorative therapies for Parkinson disease, aimed at delivering drugs or cells to the substantia nigra (SN), are becoming available. Therefore, precise surgical approaches to the dopaminergic cell-containing region of the SN are required to avoid damage to nearby structures such as the corticospinal tract and subthalamic nucleus. In a study conducted in nonhuman primates, the authors evaluated the utility and accuracy of electrophysiological techniques in localizing the SN. Three adult rhesus monkeys were used as hosts for intranigral cell transplants. The monkeys were rendered hemiparkinsonian by intracarotid injection of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. With the aid of stereotactic guidance, chronic recording chambers were placed on the skull of each monkey and directed at the SN. In each monkey, 20 to 40 trajectories were explored with a microelectrode. Spontaneous and movement-related single-unit activities were recorded in the SN, pars reticulata, subthalamic nucleus, globus pallidus, striatum, thalamus, and red nucleus. Motor and ocular responses to microstimulation in the subthalamic area were noted. Using the electrophysiological and stereotactic information that was obtained, three-dimensional maps of the nigral complex were constructed to infer the location of the SN pars compacta. The maps were subsequently used to guide intranigral placement of fetal dopaminergic cells. Accurate delivery was verified by histological analysis. Based on the characteristic electrophysiological properties of the SN and surrounding structures in the parkinsonian state, microelectrode recording techniques may be used to ensure accurate placement of cell transplantation in the intranigral region. PMID- 11014555 TI - Use of titanium mesh for reconstruction of large anterior cranial base defects. AB - The authors evaluated the role of titanium mesh used in combination with vascularized pericranium to provide rigid support during reconstruction of anterior skull base defects. Thirteen patients with large anterior skull base defects caused by tumor invasion or traumatic injury involving the cribriform plate, orbital roof, and planum sphenoidale were included in the study. The reconstruction technique involved placement of titanium mesh between two layers of continuous vascularized pericranium. Surgical glue and routine lumbar cerebrospinal fluid (CSF) drainage were not used in any patient. At a mean postoperative follow-up time of 22 months (range 8-39 months), none of the patients had developed infection or meningocele. Postoperative CSF rhinorrhea occurred in two patients with extensive dural defects, which resolved with temporary lumbar drainage. Use of titanium mesh and a two-layer vascularized pericranial graft is a safe, reproducible, and feasible method for reconstructing the anterior skull base. Patients with large dural defects may need temporary CSF diversion to avoid postoperative fistula formation. PMID- 11014556 TI - Intracranial oculomotor nerve rhabdomyoma. PMID- 11014557 TI - Anaplastic astrocytoma invading the optic chiasm through the optic pathway. PMID- 11014558 TI - Hypothermia and severe brain injury. PMID- 11014559 TI - Effect of Guglielmi detachable coils on aneurysm management. PMID- 11014560 TI - Ependymomas in children. PMID- 11014561 TI - Nodular medulloblastoma. PMID- 11014562 TI - Cochlear region of the brainstem. PMID- 11014563 TI - Keratin pearls. PMID- 11014564 TI - Genetic classification of combined hepatocellular-cholangiocarcinoma. AB - Combined hepatocellular-cholangiocarcinoma (combined HCC/ CC) is a rare form of liver neoplasms showing both hepatocellular (HCC) and bile duct differentiation (CC). In an attempt to clarify the clonality and genetic/phenotypic relationships in the evolution of these neoplasms, we microdissected multiple HCC and CC foci and studied allelic status of chromosome arms 1p, 1q, 3p, 4q, 5q, 6q, 8p, 9p, 10q, 11q, 13q, 16q, 17p, 17q, 18q, and 22q. Overall, the highest frequency of loss of heterozygosity (LOH) was seen on 4q and 17p, followed by 8p and 16q. Of the 11 cases studied, 3 cases did not show any of the identical allelic losses between HCC and CC foci, indicating the biclonal nature. The remaining 8 cases showed multiple allelic losses shared between both components, strongly suggestive of a single clonal derivation. Moreover, 4 of the 8 cases showed additional or divergent allelic losses at more than 1 chromosomal locus only in HCC and/or CC foci. Thus, this heterogeneity was shown to affect the phenotypic diversity of the tumor. Summarizing the genetic patterns, combined HCC/CC could be classified into the following 3 possibilities: (1) collision tumor in which 2 independent neoplastic clones develop at close proximity; (2) single clonal tumor with homogeneous genetic background in both components--histological diversity is thus a manifestation of divergent differentiation potential of a single clone; (3) single clonal process in which genetic heterogeneity in the process of clonal evolution within the tumor parallels histologic diversity; therefore, the tumor in this category is mainly composed of mosaics of closely related subclones. PMID- 11014565 TI - Evaluation of telomerase activity in cutaneous melanocytic proliferations. AB - Telomerase is an enzyme which synthesizes the telomeres, TTAGGG repeats at the end of vertebrate chromosomes. Its activity is suppressed in the majority of somatic cells, whereas it is detectable in most tumor cell lines and human cancers. Telomerase activity has been evaluated in many tumors for diagnostic purposes, and an increase thereof has been found with tumor progression. In our study we used anonisotopic polymerase chain reaction (PCR)-based TRAP (telomeric repeat amplification protocol) method to quantify the level of telomerase activity in a series of cutaneous melanocytic lesions. Thirty-three benign nevi, 8 dysplastic nevi, 38 malignant melanomas, and 4 melanoma metastases were analyzed. Mean relative telomerase activity was low in benign nevi (3.5+/-2.9), and significantly increased in dysplastic nevi (13.1+/-6.8), malignant melanomas (49.8+/-29.6), and metastases (121.2+/-11.2). In addition to the evaluation of telomerase activity as a possible diagnostic tool, its increase with tumor progression also suggest a prognostic role in cutaneous melanoma. PMID- 11014566 TI - Correlation between testicular biopsies (prepubertal and postpubertal) and spermiogram in cryptorchid men. AB - Twenty-one young men who underwent testicular biopsy and orchidopexy in infancy consulted owing to infertility and had biopsies again. The first and second biopsy specimens from these patients were compared by means of a semiquantitative study of the seminiferous tubules to evaluate the evolution of germ cells and to correlate these data with spermatozoon numbers. The infant testes showing lesions were classified into 3 types according to the mean tubular diameter and tubular fertility index: (1) slight lesions, (2) marked germinal hypoplasia, and (3) severe germinal hypoplasia. In the adult testes, spermatogenesis was evaluated by calculating the average numbers of spermatogonia, primary spermatocytes, young spermatids, and mature spermatids. These testes were classified as (1) normal; (2) having lesions in the adluminal compartment; (3) having lesions in the basal compartment; and (4) mixed atrophy. The number of differentiated spermatids was correlated with the expected number of spermatozoa in the ejaculate by a power regression curve. The observation of certain histologic lesions in the seminiferous tubules was assumed to indicate excretory duct obstruction: ectasia, indented outline of the seminiferous epithelium, intratesticular spermatocele, apical cytoplasmic vacuolation of Sertoli cells, and mosaic distribution of testicular lesions. There was a correlation between the prepubertal lesions and the degree of spermatogenesis in postpubertal biopsy specimens. The evolution of the 40 testes without regard to their location in infancy (cryptorchid or scrotal) was as follows. The 14 infant testes with a normal histologic pattern (5 testes) or minor lesions (9 testes) evolved to testes with lesions of the adluminal compartment (8 testes), mixed atrophy (4 testes), or lesions of the basal and adluminal compartments (2 testes). The 6 testes with marked germinal hypoplasia evolved to testes with mixed atrophy. The 20 testes with severe germinal hypoplasia evolved to testes with mixed atrophy (17 testes), Sertoli cell-only tubules (2 testes), or lesions in the basal compartment (1 testis). In the 9 patients with a histologic pattern of obstruction bilaterally (6 men) or unilaterally (3 men), the expected number of spermatozoa according to the correlation curve was much higher than the actual number in the spermiogram. This means that the testes of many azoospermic men produce spermatozoa, and this finding corroborates the importance of testicular biopsy in infertility studies. PMID- 11014567 TI - Microsatellite alterations in differentiated-type adenocarcinomas and precancerous lesions of the stomach with special reference to cellular phenotype. AB - To elucidate the relationship between genetic alterations and cellular phenotype of differentiated-type adenocarcinomas and precancerous lesions of the stomach, we phenotyped 61 gastric tumors consisting of 33 noninvasive lesions and 28 submucosal invasive carcinomas by histochemical and immunohistochemical techniques, including analysis of mucin expression. We then analyzed loss of heterozygosity (LOH) at tumor suppressor loci, examined microsatellite instability (MSI), and compared the results according to cellular phenotype. Of the 61 gastric tumors studied, 7% (4 of 61) were classified as tumors with a gastric foveolar epithelial phenotype (foveolar-type), 8% (5 of 61) as tumors with a complete-type intestinal metaplastic phenotype (CIM-type), and the remaining 85% (52 of 61) as tumors with an ordinary phenotype (ordinary-type). Forty-two percent (26 of 61) of the tumors showed LOH on at least 1 chromosomal arm. Although LOH was rare in foveolar-type tumors, it was present at variable frequencies at each tumor suppressor loci in tumors with other cellular phenotypes. p53 overexpression was observed in 0% (0 of 4) of foveolar-type, 48% (25 of 52) of ordinary-type, and 80% (4 of 5) of CIM-type tumors. With regard to MSI, all (4 of 4) of the foveolar-type tumors were classified as having high-rate MSI (MSI-H), whereas all (5 of 5) of the CIM-type tumors were microsatellite stable (MSS). Of 52 ordinary-type tumors, 19% (10 of 52) were classified as MSI H, 12% (6 of 52) as low-rate MSI (MSI-L), and 69% (36 of 52) as MSS. The incidence of MSI-H was found to be significantly higher in foveolar-type tumors (100%; 4 of 4) than in ordinary-type (19%; 10 of 52) or CIM-type tumors (0%; 0 of 5) (P < .01). An inverse correlation between MSI-H and p53 overexpression was also noticed (P < .01). Results suggested that each cellular phenotype followed a different genetic pathway; foveolar-type tumors followed the "mutator" pathway, characterized by MSI, CIM-type tumors followed the "suppressor" pathway, characterized by LOH of tumor suppressor loci and p53 overexpression, and ordinary-type tumors appeared to show mixed genetic alterations of both types. PMID- 11014568 TI - Frequency of factor V(Leiden) and prothrombin G20210A in placentas and their relationship with placental lesions. AB - The most common hereditary hypercoaguable states are factor V(Leiden) (FVL) and prothrombin mutations (PRO). FVL and PRO present with an incidence of approximately 5% in a heterogeneous population, and 45% to 63% of the thrombophilic population. The frequency of these mutations in the fetal population and their clinical importance is unknown. Fetal side thromboembolic events (FST) include congenital stroke and renal vein thromboses. In some cases, FST can be diagnosed by placental histopathology when avascular (infarcted) villi are present in a patent maternal vascular space. FST can present as placenta fetal-vascular or fetal-visceral-vascular lesions. Causes include vascular damage from cord compression or inflammation, but most remain unclear. Potential causes of FST include FVL and PRO. We describe the incidence of FVL and PRO from a prospective group of 169 consecutive placentas and in a retrospective group of archived placentas diagnosed with placental FST. One each of FVL and PRO heterozygosity was found in the prospective set (< 1% incidence for each). Five prospective placentas were diagnosed with placental FST, for an incidence of 3%; all were wild-type for FLV and PRO. Twenty-seven of 65 archived FST cases had analyzable DNA to find 5 FVL heterozygotes (18.5%); all were wild-type for PRO. Twenty-one of 65 retrospective archived controls analyzable found 1 case of FVL heterozygosity (< 5%). We find that the frequency of FVL and PRO may be decreased in the pregnant population but increased in cases of placental FST. Because factor V Leiden heterozygosity carries an increased risk for thrombotic complications, we suggest placental diagnosis of fetal side thromboemboli warrants clinical evaluation for FVL in infant and potentially the parents. PMID- 11014569 TI - Pathology residents' use of a Web-based tutorial to improve Gleason grading of prostate carcinoma on needle biopsies. AB - Little is known about pathology residents' ability to Gleason grade or their ability to learn surgical pathology using Internet-based technology. A free Web based program (available at www.pathology. jhu.edu/prostate) was developed that consisted of 20 pretutorial images for grading, 24 tutorial images, and the same 20 posttutorial images for Gleason grading. The grading images were selected from cases that had a consensus Gleason grade from 10 uropathology experts. In 2.5 months, 255 residents visited the website, and 151 (59%) completed it. Of those who completed the website, their year in training was known in 85 (56%): 1st year, 25.8%; 2nd year, 20%; 3rd year, 22.3%; 4th year, 14.1%; 5th year, 15.3%; and 6th year, 2.4%. Eighty percent learned Gleason grading in residency versus being self-taught, and 66% were male. In a multivariate analysis, higher pretutorial scores were associated with both their year in training (P = .001) and their hospital size (P = .003). Improvements in grading posttutorial were not related to the residents' year in training. Overall, the website significantly improved grading in 11 of 20 images and had no effect in 9 of 20 images. Improvements were noted in 1 of 1 Gleason score 4; 2 of 7 Gleason score 5 to 6; 2 of 6 Gleason score 7; and 6 of 6 Gleason score above 7 tumors. In summary, a Web based tutorial improved Gleason grading accuracy by pathology residents to an equal extent regardless of their year in training. It is more difficult to teach residents to grade Gleason scores 5 to 7 tumors, and additional training should be concentrated in this area. PMID- 11014570 TI - Anti-CD10 immunoperoxidase staining of paraffin-embedded acute leukemias: comparison with flow cytometric immunophenotyping. AB - CD10 is common in B-precursor acute lymphoblastic leukemia (ALL) but is rare in acute myeloid leukemia (AML). However, until recently, analysis for CD10 has generally required fresh or frozen tissue. 56C6 is a monoclonal antibody that is now commercially available for the detection of CD10 in routinely processed paraffin-embedded tissue. Immunoperoxidase stains for CD10 on paraffin-embedded bone marrow core biopsy specimens (B5-fixed, decalcified) and marrow aspirate clots (formalin-fixed) were compared with flow cytometric immunophenotyping for CD10 on fresh cell suspensions in 20 cases of AML and in 30 cases of ALL. CD10 detection by immunohistochemistry agreed with CD10 by flow cytometry in 98% (49 of 50) of acute leukemias. The results matched in 100% (20 of 20) of AML. Five percent (1 of 20) of AMLs expressed CD10. Two of the AMLs with monocytoid differentiation were interpreted as negative for CD10 by flow cytometry, although these had nonspecific dim immunofluorescence for multiple markers, including CD10, and these cases were negative by immunohistochemistry. CD10 detection by immunohistochemistry agreed with CD10 by flow cytometry in 97% (29 of 30) of ALL. Eighty-four percent (21 of 25) of B-precursor ALL and 40% (2/5) of T-lineage ALL expressed CD10 by immunohistochemistry. In 1 case of B-precursor ALL, CD10 was dimly positive in 24% of the blasts by flow cytometry but negative by immunohistochemistry. We conclude that immunohistochemical staining of paraffin embedded tissue, either B5- or formalin-fixed, is an effective method for the detection of CD10 in acute leukemia. This technique is useful in distinguishing AML from ALL. PMID- 11014571 TI - Inhibin-alpha CD99, HEA125, PLAP, and chromogranin immunoreactivity in testicular neoplasms and the androgen insensitivity syndrome. AB - We investigated 115 testicular and 3 epididymal tumors and 6 cases of the complete androgen insensitivity syndrome (AIS) for the expression of inhibin alpha, CD99, HEA125, PLAP, and chromogranin, using monoclonal antibodies and standard immunhistochemical techniques. Ihibin-alpha was detected in the neoplastic cells in 27 of 27 primary Leydig cell tumors (LCTs), 1 of 1 metastatic LCT, 6 of 20 Sertoli cell tumors (SCTs), 4 of 5 juvenile granulosa cell tumors (GCTs), and 2 of 5 unclassified sex cord-stromal tumors (USCSTs). Except for 2 choriocarcinomas, the choriocarcinomatous component of 1 mixed germ cell tumor, and a small focus of inhibin-positive syncytiotrophoblast in 1 embryonal carcinoma, inhibin-a immunoreactivity was not present in the neoplastic cells of the 38 remaining testicular germ cell tumors; 11 B-cell and 1 T-cell lymphomas; 1 granulocytic sarcoma; and 1 rhabdomyosarcoma of the testis; 1 adenoma of the rete testis, and 3 adenomatoid tumors of the epididymis. Inhibin-alpha immunoreactivity was present in the Sertoli cells and Leydig cells in 5 testicular hamartomas and in 1 Sertoli cell adenoma in 6 cases of AIS; both Sertoli and Leydig cells were also positive in the extranodular testicular parenchyma present in 2 of these cases. CD99 was detected in 10 of 15 primary LCTs, 1 of 7 SCTs, 3 of 5 JGCTs, and in 1 of 5 USCSTs but was not found in any tumor outside the sex cord-stromal category. HEA125 immunostaining was not detected in sex cord-stromal tumors; however, 3 of 12 seminomas, 3 of 12 embryonal carcinomas, 6 of 8 yolk sac tumors, and 1 of 2 teratomas were HEA125 positive. PLAP was not detected in sex cord-stromal tumors except for 4 of 15 primary LCTs but was present in most germ cell tumors. Chromogranin immunostaining was present in the sex cord-like element in 1 of 5 USCSTs, 1 of 8 YSTs, 1 of 2 teratomas, and in 1 of 1 rete adenoma, and in normal adjacent rete testis. In conclusion, although inhibin-alpha and PLAP, and, to a somewhat lesser extent, CD99 and HEA125 immunostaining are helpful in the differential diagnosis of certain testicular neoplasms that are difficult to distinguish on morphologic grounds, chromogranin is far less helpful in this context. PMID- 11014572 TI - Distribution of keratins in normal endothelial cells and a spectrum of vascular tumors: implications in tumor diagnosis. AB - Vascular endothelial cells are specialized mesenchyme-derived epithelial-like lining cells which are the essential participants in benign and malignant vascular tumors. Although endothelia in lower animals often express keratins (K), human endothelia are generally K negative and vimentin-positive. However, K expression has been noted in some endothelia and in some epithelioid vascular tumors. In this study, we systematically examined normal human vascular endothelia and a spectrum of human vascular tumors (n = minimum of 137 tumors with each marker) for simple epithelial keratin polypeptides of the Moll catalogue (K7, K8, K18, and K19). Selected vascular tumors were also evaluated with antibodies to K14 and the monoclonal antibody 34betaE12 that recognizes several keratins of stratified epithelia. Endothelia of normal veins, venules, and lymphatics commonly exhibited focal positivity for K7 and K18, whereas K8, K14, and K19 were not seen in non-neoplastic endothelia with the antibodies used. Lymphangiomas (6 of 7) and venous hemangiomas (6 of 13) often showed K7-positive endothelial cells; K18 was detected less commonly, whereas K8 and K19 were not detected. Epithelioid hemangioendotheliomas (EHEs) showed K7 and K18 expression in the majority of cases (50% and 100%, respectively), while K8 was seen in 10% cases and K14 and K19 in none. In contrast, epithelioid angiosarcomas (EAs) were often positive for K8 and K18 (approximately 50%), whereas they less commonly showed K7 and only occasionally K19; all tumors were negative for K14 and with the antibody 34betaE12. Nonepithelioid angiosarcomas (AS) less commonly showed keratin expression with K7, K8, and K18 being positive in 20% of cases, and K14 and K19 in none of the cases. Epithelial membrane antigen (EMA) was occasionally detectable in EHE (2/19) but was present in 4 of 16 (25%) EAs and 17 of 48 (35%) nonepithelioid AS. These findings document the common presence of focal reactivity for K7 and K18 in subsets of normal endothelia and also the frequent presence of simple epithelial keratins in malignant vascular tumors, while such expression is uncommon in nonepithelioid angiosarcomas. K- and EMA-positivity in neoplastic endothelia needs to be considered in the evaluation of human tumors. K antibodies such as those specific to K19 or AE1 that do not react with K8 and K18 should be used in the differential diagnosis of epithelioid vascular tumors and carcinomas. PMID- 11014573 TI - The expression of fatty acid synthase (FASE) is an early event in the development and progression of squamous cell carcinoma of the lung. AB - Correlation of elevated levels of the lipogenic enzyme, fatty acid synthase (FASE), with advanced stages of some cancers has drawn attention to this enzyme as a possible marker of poor prognosis. Because recent studies have shown that cancer cells are dependent on fatty acid synthetic activity and pharmacologic inhibitors of this enzyme are selectively cytotoxic to cancer cells, expression of FASE also may provide a potential target for intervention in the neoplastic process. To determine the potential usefulness of expression of FASE in the neoplastic process of the lung, we evaluated its pattern of expression immunohistochemically in archival specimens from 60 human lung specimens with squamous cell cancer (SCC) and associated "preneoplastic" lesions compared with its expression in the normal bronchial epithelium of 60 noncancer specimens. The expression of FASE was significantly higher in SCC associated uninvolved bronchial epithelium (mean = 0.40+/-0.03, median = 0.38) compared with its expression in the bronchial epithelium of noncancer specimens (mean = 0.18+/ 0.02, median = 0.16) indicating its early expression. We also observed a statistically significant step-wise increase in FASE expression from SCC associated uninvolved bronchial epithelium (mean = 0.40+/-0.03, median = 0.38) to epithelial hyperplasia (0.58+/-0.04, median = 0.57) to SCC (1.53+/-0.06, median = 1.50). The results suggested that expression of FASE is an early event in the development and progression of SCC of the lung. The inhibition of fatty acid synthesis by inhibiting enzymatic function with metabolic analogues may be a useful strategy in the treatment of SCCs. The expression of FASE in early lesions such as SCC associated uninvolved bronchial epithelium and epithelial hyperplasia might also provide a potential means for intervention early in the neoplastic process in the lung or even preventing their malignant transformation to invasive carcinomas. PMID- 11014574 TI - Adult rhabdomyoma of the extremity: a case report and review of the literature. AB - The adult rhabdomyoma is a rare, benign skeletal muscle neoplasm that usually occurs in the head and neck. A case report of an adult rhabdomyoma arising in the thigh is presented with a review of the literature. This is the first case of an extremity adult rhabdomyoma to be reported. It is also the largest at 13 centimeters. Distinction from a highly differentiated rhabdomyosarcoma is important. Recent chromosomal studies suggest that the adult rhabdomyoma is a true neoplasm. Total resection is curative but the lesion may recur if incompletely excised. PMID- 11014575 TI - Carbohydrate antigen expression in primary tumors, metastatic lesions, and serous effusions from patients diagnosed with epithelial ovarian carcinoma: evidence of up-regulated Tn and Sialyl Tn antigen expression in effusions. AB - The object of this study was the investigation of carbohydrate antigen expression in malignant epithelial cells and benign mesothelial cells in serous effusions from patients diagnosed with epithelial ovarian carcinomas. In addition, to compare antigen expression in carcinoma cells in effusions with those of corresponding primary tumors and metastatic lesions. Sections from 63 malignant effusions from ovarian carcinoma patients and 15 reactive effusions were immunohistochemically stained, using 5 monoclonal antibodies for Lewis(y), Sialyl Lewis(x), Tn, and Sialyl Tn antigens. Tissue sections (n = 97) from corresponding primary ovarian carcinomas and metastatic lesions, as well as from 12 malignant mesotheliomas, were additionally stained using the above panel. Staining for the 4 antigens was seen in carcinoma cells in serous effusions in the majority of cases (range = 71% to 85%). In contrast, immunoreactivity was detected in mesothelial cells in only 6% to 23% of the specimens studied (P < .001 for all 5 markers). With the exception of B3 antibody against Lewis(y) antigen, malignant mesotheliomas stained negative, infrequently showing focal immunoreactivity. An up-regulation of Tn and Sialyl Tn expression was detected in carcinoma cells in effusions when compared with both primary tumors (P < .003 and P < .007, respectively) and metastatic lesions (P < .034 and .041, respectively). Cancer associated carbohydrate antigens can thus be used as an adjunct in the differentiation between malignant epithelial and reactive mesothelial cells. Ovarian carcinoma cells in effusions show up-regulation of Tn and Sialyl Tn, possibly representing a transient phenotypic alteration facilitating metastasis. PMID- 11014576 TI - Reduced expression of CD44v3 variant isoform is associated with unfavorable outcome in non-small cell lung carcinoma. AB - The expression of CD44 standard form (CD44s) and variant isoforms v3 and v6 was analyzed in 233 resected non-small cell lung carcinoma (NSCLC) specimens by immunohistochemistry (IHC), and the mRNA status of CD44v3 and CD44v6 in a cohort of samples was determined by in situ hybridization (ISH) and further confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). The expression of CD44s, CD44v3, and CD44v6 was correlated with clinicopathologic variables and survival. The expression of CD44v3 and v6 was reduced in 97% and 90% of the adenocarcinomas and in 86% and 74% of the large cell/anaplastic carcinomas, respectively, as compared with squamous cell carcinomas, where they were reduced in 53% and 51% of the cases (P = .0001 and P = .004 for v3 and v6). The corresponding values for CD44s were 92%, 70%, and 51%, respectively (P = .011). The reduced CD44s and CD44v6 expression was associated with lymph node metastases (P = .03 and P = .005, respectively) and the reduced expression of CD44s also with advanced stage (P = .04). Recurrences during the follow-up were more often found within the tumors showing reduced expression of CD44v3 (P = .04). Combining ISH and IHC results showed that CD44v3 and v6 mRNA were not always processed into protein, suggesting a regulation disturbance posttranscriptionally since malignant transformation of cells has occurred. In survival analyses, the reduced expression of CD44s and CD44v3 was associated with a shortened disease-free survival (P = .04 and P = .01, respectively). In multivariate analysis, CD44v3 retained its independent prognostic value (P = .03). These results emphasize the value of CD44, and especially the v3 variant isoform in the behavior of NSCLC. PMID- 11014577 TI - CK7 expression in carcinomas of the Waldeyer's ring area. AB - Primary carcinomas of the Waldeyer's ring area are typically nonkeratinizing squamous cell carcinomas (SCC). Their cervical lymph node metastases are not uncommonly cystic and filled with necrotic tumor cells. Some cysts, however, contain clear fluid. During the investigation of SCC producing "fluid-filled" cystic metastases, we evaluated hematoxylin and eosin (H&E) sections of 90 primary SCC for their site of origin. We analyzed the cytokeratin (CK) profile of primary and metastatic carcinoma with special focus on the expression of CK7, a putative marker for ductal differentiation. CK7 was expressed in submucosal minor salivary gland acini and ducts, but not in the squamous surface epithelium of the Waldeyer's ring. CK7 was expressed in 11 primary SCC (8 base of tongue/3 palatine tonsil). The CK7-positive SCC were deep-seated, arose from large excretory ducts of submucosal minor salivary glands, and showed only insignificant surface involvement. They were characterized by a solid infiltrative growth pattern of basaloid cells with focal ductal differentiation. Salivary ducts adjacent to the carcinoma showed extensive intraductal hyperplasia and metaplasia. All CK7 positive carcinomas produced CK7-positive cystic nodal metastases, most of which contained paucicellular fluid. No solid CK7-positive nodal metastases were identified. In summary, a subset of carcinomas occurring in the Waldeyer's ring area appear to arise from large excretory ducts of submucosal minor salivary glands with only limited surface involvement, express CK7, and produce CK7 positive cystic "fluid-filled" nodal metastases. The histomorphology and immunophenotype suggest that these carcinomas represent basaloid SCC arising from excretory ducts of the submucosal minor salivary glands. PMID- 11014578 TI - Diagnostic effect of an improved preembedding method of prostate needle biopsy specimens. AB - The authors compared the influence of a conventional and an optimized submitting method of prostate core needle biopsy specimens on the frequency of cancer detected and the pathologic characteristics of the adenocarcinoma bearing biopsy specimens. The patients included were part of the prostate-specific antigen (PSA) screening program of Tyrol/Austria. Of the systematic core needle biopsy specimens from 500 unselected men obtained within 1 year from the Urological Department, University of Innsbruck, the core biopsy specimens of 250 cases were submitted conventionally, floating free in formalin-filled containers, whereas the biopsy specimens of the other 250 cases were stretched and orientated between 2 meshes in tissue cassettes at the time of biopsy before formalin fixation. On 136 cases diagnosed as adenocarcinoma the number and the length of cores as well as number of the cores involved by cancer and the tumor size were morphometrically determined. The diagnosis of benign prostatic hyperplasia, isolated high-grade prostatic intraepithelial neoplasia (PIN), atypical foci suspicious for cancer, and carcinoma was made in 66%, 5.6%, 4.8%, and 23.6% after conventional submission and in 61.6%, 6.4%, 1.2%, and 30.8% of the cases after optimized preembedding respectively. In the adenocarcinoma cases the optimizedly preembedded material showed higher mean total core length (126.5 mm versus 93.9 mm; P < .0001), a higher mean total tumor length (14.1 mm versus 8.6 mm; P = .01), and more cores involved by cancer (2.9 versus 2.4; P = .01) compared with the conventionally worked-up biopsy specimens. Optimized preembedding of core needle biopsy specimens in tissue cassettes could be quickly and routinely done by the assistance of the urologists at the time of biopsy. The significant improvement of the histologic yield of optimizedly preembedded prostatic needle biopsy specimens led to a higher frequency of cancer diagnosis, a reduction of cases with atypical foci suspicious for cancer and a significantly lower number of cases with only 1 core biopsy involved by cancer. PMID- 11014579 TI - Lipomatous hemangiopericytoma: a fat-containing variant of solitary fibrous tumor? Clinicopathologic, immunohistochemical, and ultrastructural analysis of a series in favor of a unifying concept. AB - The dinicopathologic, immunohistochemical and ultrastructural features of 13 lipomatous hemangiopericytomas are presented. There were 6 male and 7 female patients whose ages at diagnosis ranged from 27 to 75 years (median 48) all presenting with a mass of variable duration. The tumor sizes ranged from 1.7 cm to 19 cm (median 5.5 cm). The locations included the orbit (1), neck (1), mediastinum (1), epicardium (1), retroperitoneum (3), right iliac fossa (1), and upper (1) and lower (4) extremity. Histologically, the lesions were composed of a varying admixture of spindle-shaped to round cells, variably collagenous stroma, adipose tissue, and branched, often thick-walled, hemangiopericytoma-like vessels. For 11 tumors, the mitotic activity ranged from 1 to 3 mitoses per 10 high-power fields (HPF). One tumor which contained hypercellular areas showed 13 mitoses per 10 HPF, and another hypercellular lesion showed up to 43 mitoses per 10 HPF, abnormal mitoses, and necrosis. Immunohistochemically, tumor cells were invariably positive for vimentin and CD99, and mostly for CD34 but negative for desmin, keratin, CD31, CD117 (c-kit), and inhibin. About half of the tumors showed reactivity for bcl-2. Occasionally, focal reactivity was also observed for smooth muscle actin, muscle-specific actin, S100 protein, and epithelial membrane antigen. Ultrastructural examination of seven cases showed features in keeping with fibroblastic, myofibroblastic, or pericytic differentiation. Treatment consisted of simple tumorectomy in 10 cases and wide excision in 3. Follow-up information on 10 patients (range: 6 to 77 months; median: 18 months) showed no recurrence. Lipomatous hemangiopericytoma which share the clinical, pathologic, immunohistochemical, and ultrastructural features of solitary fibrous tumor (SFT) is likely to represent, in most cases, a fat-containing variant of SFT. PMID- 11014580 TI - Absence of RBM expression as a marker of intratubular (in situ) germ cell neoplasia of the testis. AB - RBM (RNA-binding motif) protein is a marker of male germ cells. This protein is encoded by the Azoospermia factor region-b (AZF-b) of the human Y chromosome and is expressed exclusively in the male germ cell line, that is, spermatogonia, spermatocytes, and round spermatids. The authors analyzed the expression of the RBM gene in germ cell tumors and in the seminiferous tubules in the vicinity of these tumors to identify the presence of IGCN. Sections from testicular germ cell tumors of 21 patients were stained with anti-RBM antibody by using an immunohistochemical method. Distal tubules showing spermatogenesis were immunopositive for RBM protein. All of the germ cell tumors studied were completely immunonegative for RBM. Defined areas of IGCN also showed an absence of RBM expression. Tubules with spermatocyte-like cells, which were expected to express RBM, did not express this protein. This result enabled the identification of tubules as being IGCN. RBM is a novel marker consistently expressed in normal male germ cells but not in malignant germ cell tumors or IGCN. Thus, the absence of RBM expression in germ cells provides a new diagnostic tool of preinvasive malignancy of the testis. PMID- 11014581 TI - Acute thymic involution in fetuses and neonates with chorioamnionitis. AB - Chorioamnionitis represents the leading cause of preterm birth and related pathologic conditions as well as of fetal death and frequently occurs in symptom free mothers. Recent radiologic findings have indicated that thymus size is significantly reduced in preterm infants born to mothers with subclinical, histologically proven chorioamnionitis. However, an accurate morphologic description of the thymus gland in fetuses and neonates with chorioamnionitis is lacking, although it is known that infection and other stress processes may cause lymphocyte depletion in the thymuses of infants and older babies (acute stress involution). We describe morphologic modifications in the thymus of fetuses with histologically proven chorioamnionitis and newborn infants with chorioamnionitis and proven sepsis. The main findings included (1) decreased organ volume (ANOVA, P < .0024); (2) reduced corticomedullary ratio (P < 10(-6)); (3) significant changes in the relationship between thymic parenchyma and thymic interstitial tissue with resulting increased organ complexity (P = .03); (4) severe reduction of thymocytes; and (5) other degenerative processes such as monocyte/macrophage infiltration of Hassall's bodies. These results indicate that chorioamnionitis, with or without sepsis, is associated with significant morphologic modifications in the thymus. We wish to note that the described thymic pathology is only one aspect of the fetal systemic inflammatory response syndrome with which chorioamnionitis is associated. PMID- 11014582 TI - Peritubular capillaries in chronic renal allograft rejection: a quantitative ultrastructural study. AB - Peritubular capillaries (PCs) with a circumferentially multilayered basement membrane have been suggested as an ultrastructural indicator of chronic renal allograft rejection (CR). The authors validated this lesion as a marker for CR, by analyzing its quantitative features, specificity, and sensitivity in 169 renal biopsy specimens. The mean number of circumferential layers (PCcirc) and the incidences of the grades (mild: 2 to 4, moderate: 5 to 6, severe: 7 or more layers) were investigated in biopsy specimens involving CR (CR(Bx), n = 46), acute rejection (n = 11), normal kidneys (n = 20), psoriatics treated with cyclosporine (n = 13), renal transplants with chronic cyclosporine toxicity (n = 12), native kidney diseases (NKD, n = 56), and transplant nephrectomies attributable to CR (Cr(nephr), n = 11). CR was diagnosed with regard to the clinical features and the presence of intimal fibrosis in 41 biopsy specimens or transplant glomerulopathy in 35 biopsy specimens (cg; identified only by electron microscopy in 10 cases). NKD included chronic glomerulonephritis, chronic tubulointerstitial nephritis, benign nephrosclerosis, thrombotic microangiopathy, diabetic nephropathy, and renal disease in elderly patients (median age, 72 years). All PCs around glomeruli were sampled (median, 14 profiles per case). PCs with a moderate/ severe lesion appeared as serrated profiles with a thick, ribbon like basement membrane layer in semithin plastic sections. The numbers of circumferentially multilayered PCs were significantly characteristic of CR (PCcirc in CR(Bx): 2.87+/-1.83 SD; range, 0 to 7.36; P < .001 v other groups). A severe lesion occurred exclusively in CR (in 12% of the PCs in CR(Bx), and in 38% in CRnephr). A moderate lesion was observed in 0.6% of the PCs in NKD, 16% in CR(Bx), and 21% in CRnephr. Three or more PCs with a moderate lesion were encountered only in CR. A mild lesion was not suggestive of CR at all. In CR(Bx), 27 cases showed a severe lesion or 3 or more PCs with a moderate lesion (cpc; sensitivity: 59%). Four of the 27 cases lacked cg. The cumulative incidence of cpc and cg was 85%. In transplants with cyclosporine toxicity, the presence of cpc verified the coexistence of CR in 7 specimens. In conclusion, cpc is a specific marker of CR. The incidence of cpc increases as CR progresses. The lesion may be caused by a low-grade rejection injury to the PCs. Careful analysis of semithin sections promotes the better sampling of cpc. An ultrastructural demonstration of cpc and cg defines CR more precisely than does light microscopic evaluation per se. PMID- 11014583 TI - Variable expression of keratins and nearly uniform lack of thyroid transcription factor 1 in thyroid anaplastic carcinoma. AB - Thyroid anaplastic (undifferentiated) carcinomas (TACs) comprise a morphologically heterogeneous group of tumors, which can arise in the background of differentiated papillary or follicular carcinoma. The thyroid epithelial differentiation varies in these tumors and has not been completely characterized. In this study, we immunohistochemically analyzed different variants TACs from 35 patients by using antibodies specific to 9 different keratin polypeptides, epithelial membrane antigen, thyroid transcription factor I (TTF-1), and thyroglobulin. These tumors were histologically divided into 3 categories: squamoid-cohesive (SC, 13 tumors), spindle cell sarcomatous (SS, 8 cases) and intermediate group, including tumors with giant cells and solid epithelioid components (GC, 18 tumors); 4 tumors had 2 components. The patients ages ranged from 40 to 89 years, with a mean age in all groups of 70 years. TTF-1 was present in only 2 of 9 of the SC tumors, and absent in all other TACs, but was present in entrapped differentiated components. Thyroglobulin was absent in all but 1 case. A complex keratin (K) pattern of stratified epithelia was typically seen in the SC tumors with extensive K7, K8, K17, K18, and K19, and variable K13 and K14 expression; EMA was also present. K16 was limited to squamous pearls in 1 tumor, and K10 was absent. The GC carcinomas typically had K8 and K18, whereas the expression of K7 was variable and that of K14, K17, and K19 sporadic; EMA was variably present in half of the cases. The keratins in spindle cell sarcomatous tumors were usually limited to K7, K8, and K18, often in limited numbers of cells. EMA was present in 1 case only. These results indicate a complex pattern of keratins in squamoid and giant cell TACs, similar to papillary carcinoma and suggesting the possibility of relationship. There was a progressive loss of epithelial differentiation and keratins in sarcomatoid TACs. Loss of TTF-1 is a nearly uniform feature of TAC and disallows the use of this marker to pinpoint a thyroid origin of these tumors. PMID- 11014584 TI - Reduced 15-lipoxygenase-2 immunostaining in prostate adenocarcinoma: correlation with grade and expression in high-grade prostatic intraepithelial neoplasia. AB - Arachidonic acid (AA) metabolites are implicated in the oncogenesis of several tumors, including prostate cancer. 15-Lipoxygenase-2 (15-LOX-2) is a novel AA metabolizing enzyme with a limited tissue distribution, which includes prostate, lung, skin, and cornea. Previous studies have shown that 15-LOX-2 is present in benign prostate secretory cells and reduced in prostate adenocarcinoma and that production of the 15-LOX-2 metabolite 15S-hydroxyeicosatetraenoic acid is reduced in malignant compared with benign prostate. The objective of this study was to determine the frequency with which 15-LOX-2 immunostaining is reduced in prostate carcinoma and to correlate reduced expression with tumor differentiation (grade) and other pathologic parameters in radical prostatectomy specimens. Paraffin immunoperoxidase with a polyclonal antibody specific for 15-LOX-2 was performed on tumors and benign portions from 70 cases, and the percentage of tumor immunostaining for 15-LOX-2 was assessed. Whereas uniform 15-LOX-2 immunostaining was observed in secretory cells of benign glands, it was markedly reduced or absent in most adenocarcinomas: 23 of 70 tumors showed completely absent 15-LOX-2 immunostaining, and 45 of 70 cases showed negative immunostaining in more than 50% of the tumor. The extent of reduced 15-LOX-2 immunostaining correlated with tumor differentiation, with retained expression particularly in Gleason score 5 tumors versus a significant reduction of 15-LOX-2 in higher-grade tumors (mean +/ SD tumor 15-LOX-2 positive: Gleason score 5 = 67%+/-30%, Gleason score 6 = 16%+/ 30%, Gleason score 7 = 23%+/-28%, Gleason score > or =8 = 41%+/-46%). In 16 cases with multifocal tumors or different foci of the same tumor with different grades, the higher-grade foci had significantly reduced 15-LOX-2 expression compared with the lower-grade foci. In peripheral zone tumors without complete loss of 15-LOX-2 expression, there was a significant inverse relationship between 15-LOX-2 immunostaining and tumor volume. There was not a significant correlation between 15-LOX-2 immunostaining and serum PSA or pathologic stage. In a subset of 27 cases, 15-LOX-2 expression in high-grade prostatic intraepithelial neoplasia (HGPIN) glands was significantly reduced compared with benign glands. These data show that in contrast to the uniform expression of 15-LOX-2 in differentiated secretory cells of benign prostate, reduced 15-LOX-2 is a common alteration in prostate carcinoma, and this correlates with tumor cell differentiation. That reduced expression is seen in HGPIN suggests that this may be an early alteration in carcinoma development. PMID- 11014585 TI - Efficacy of restaining prostate needle biopsies with high-molecular weight cytokeratin. AB - Prostate tissue and lesions obtained by needle biopsy may be scant and not survive cutting into the block; this study examined the efficacy of destaining hematoxylin and eosin-stained sections and restaining the slides using immunohistochemistry with high-molecular weight cytokeratin (high-molecular weight cytokeratin). We identified 105 prostate needle biopsies referred to Johns Hopkins Hospital in an 18-month period (January 1997-June 1998) that had been destained and restained for high molecular weight cytokeratin. The slides were reviewed for the Johns Hopkins Hospital diagnosis (benign, malignant, or equivocal), which had factored in the immunohistochemistry results, and for immunohistochemistry staining quality (optimal, suboptimal, stain failed, or lesion fell off). We obtained data on 96 cases from the referring institutions about the fixative and glass slides used for processing the needle biopsy. In 58% of cases, destaining and restaining with high-molecular weight cytokeratin allowed a definitive benign or malignant diagnosis to be made; in 79% of these cases, the staining was optimal. In only 13% of cases the diagnosis remained equivocal; of these, the stain worked optimally in only 36%. In 19% of cases, the stain failed. In 9% of cases, the lesion fell off; in all 7 cases with available data the tissue had been cut on non-charged slides. All but 3 cases were received in 10% neutral buffered formalin. There was no correlation between the use of charged (plus or lysine coated) or non-charged slides and the staining quality. Furthermore, in 12 instances, we received more than 1 specimen from the same referring institution, and in 6 of these instances there was variable staining in the different cases from the same institution. Destaining hematoxylin and eosin stained slides and restaining for high-molecular weight cytokeratin is a useful technique that in the majority of cases enables a definitive diagnosis to be made. Tissue may survive the procedure better if originally cut on charged slides, but staining quality is no different for charged or non-charged slides. PMID- 11014586 TI - Massive localized lymphedema: additional locations and association with hypothyroidism. AB - We report the second series of a new entity called "massive localized lymphedema in morbidly obese patients" (MLL), recently described in medical literature. Our 6 cases present additional locations as well as an association with hypothyroidism. Huge masses, of longstanding duration ranging from 9 months to 8 years, afflicted the thigh, popliteal fossa, scrotum, suprapubic and inguinal region, and abdomen of morbidly obese adults. Although clinical impressions were generally of a benign process, including lipoma and recurrent cellulitis, the possibility of a malignant neoplasm could not be eliminated. Poorly defined and non-encapsulated, these skin and subcutaneous lesions were most remarkable for their sheer size, measuring 50.6 cm in mean diameter (range, 38-75 cm) and weighing a mean of 6764.5 g (range, 2,060-12,000 g) The overlying skin exhibited the induration and peau d'orange characteristic of chronic lymphedema. Grossly and histologically, a prominent marbled appearance, rendered by fibrous bands intersecting lobules of adipose tissue, simulated sclerosing well differentiated liposarcoma. However, the absence of atypical stromal cells, atypical adipocytes, and lipoblasts precluded the diagnosis of well differentiated liposarcoma. Instead, reactive features, encompassing lymphatic vascular ectasia, mononuclear cell infiltrates, fibrosis, and edema between the collagen fibers, as well as ischemic changes including infarction and fat necrosis, established the diagnosis of MLL. Although the pathogenesis of MLL may be as simple as obstruction of efferent lymphatic flow by a massive abdominal pannus and/or prior surgery, the presence of hypothyroidism in 2 of our patients suggests an alternative pathogenesis. Recognition of this entity by both clinicians and pathologists should avert a misdiagnosis as a low-grade liposarcoma. PMID- 11014587 TI - A variant form of Churg-Strauss syndrome: initial temporal non-giant cell arteritis followed by asthma--is this a distinct clinicopathologic entity? AB - The clinical manifestations of the classical vasculitis syndromes are extraordinarily heterogenous with considerable overlap among them. Recently, several cases of unusual presentation of the vasculitis syndromes have been reported. We describe a patient who initially manifested with temporal arteritis and Raynaud's phenomenon and subsequently developed bronchial asthma, ie, a case of an atypical form of Churg-Strauss syndrome (allergic angiitis and granulomatosis) and discuss whether this case is a distinct clinicopathological entity. PMID- 11014588 TI - CD138-positive and Kaposi's sarcoma-associated herpesvirus (KSHV)-negative B-cell lymphoma with serosal spreading of the body cavity and lymphadenopathy: an autopsy case. AB - CD138-positive and Kaposi's sarcoma-associated herpes virus (KSHV)-negative B cell lymphoma with serosal spreading of the body cavity and lymphadenopathy is presented. Our lymphoma cells showed pleomorphic morphology and a clonal immunoglobulin gene rearrangement. Immunophenotypically, they lacked B- and T cell-associated antigens but expressed strong membranous CD138 antigen along the serosa. Although our case was not conventional primary effusion lymphoma (PEL) because of the absence of KSHV and the presence of lymphadenopathy, its unique phenotype and serosal spreading were consistent with those of PEL. Our case suggests that, irrespective of KSHV infection, some pleomorphic B cell lymphomas with membranous CD138 expression show a peculiar serosal spreading. PMID- 11014589 TI - Adhesion molecule detection in a case of early cerebral malaria: immunohistochemical and electron microscopic findings. AB - A case of early cerebral malaria caused by Plasmodium falciparum was studied. P selectin glycoprotein ligand 1 (PL1) was detected along the inner surface of the infected red blood cells (IRBCs), which ordinarily are not positive for PL1 immunohistochemically, suggesting PL1 being the product of parasite. The electron microscopic finding showed granular deposits in the corresponding lesion, consistent with PL1 deposition, in the IRBCs firmly attached to the endothelium of small cerebral vessels. Most of the IRBCs were round shaped as though they lost their capacity to change shape. The therapeutic strategy was expected against adhesion molecules such as PL1 and for maintaining or restoring the metamorphic capacity of IRBCs. PMID- 11014590 TI - Atypical immature metaplastic-like proliferation of the cervix. PMID- 11014591 TI - The adhesion molecule expression pattern of basal cell carcinoma cells should be compared with outer root sheath cells of hair follicle and not with stratum basale of skin. PMID- 11014592 TI - DipoCoup: A versatile program for 3D-structure homology comparison based on residual dipolar couplings and pseudocontact shifts. AB - A program, DipoCoup, is presented that allows to search the protein data bank for proteins which have a three dimensional fold that is at least partially homologous to a protein under investigation. The three dimensional homology search uses secondary structure alignment based on chemical shifts and dipolar couplings or pseudocontact shifts for the three dimensional orientation of secondary structure elements. Moreover, the program offers additional tools for handling and analyzing dipolar couplings. PMID- 11014593 TI - The auto-orientation in high magnetic fields of oxidized cytochrome b562 as source of constraints for solution structure determination. AB - 15N-1H 1J couplings were measured at 500 MHz and 800 MHz for 15N enriched oxidized cytochrome b562 from E. coli. The magnetic field dependence of 70 1J values, which could be measured without signal overlap, shows that there is a molecular magnetic anisotropy which provides partial molecular orientation in the magnetic field and, consequently, residual dipolar couplings (rdc). The rdc were used as further constraints to improve the existing structure [Arnesano et al. (1999) Biochemistry, 38, 8657-8670] with a protocol which uses the rhombic anisotropy [Banci et al. (1998) J. Am. Ctherz. Soc., 120, 12903-12909]. The overall large molecular magnetic anisotropy has been found to be determined by both the low spin iron (III) and the four helix bundle structure magnetic susceptibility anisotropy contributions. PMID- 11014594 TI - NMR detection of side chain-side chain hydrogen bonding interactions in 13C/15N labeled proteins. AB - We describe the direct observation of side chain-side chain hydrogen bonding interactions in proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the guanidinium nitrogen 15Nepsilon of arginine 71, which serves as the hydrogen donor, and the acceptor carboxylate carbon 13CO2gamma of aspartate 100 in a 12 kDa protein, human FKBP12, is detected via the trans-hydrogen bond 3h JNepsilonCO2gamma coupling by employing a novel HNCO-type experiment, soft CPD-HNCO. The 3h JNepsilonCO2gamma coupling constant appears to be even smaller than the average value of backbone 3h JNC' couplings, consistent with more extensive local dynamics in protein side chains. The identification of trans-hydrogen bond J-couplings between protein side chains should provide useful markers for monitoring hydrogen bonding interactions that contribute to the stability of protein folds, to alignments within enzyme active sites and to recognition events at macromolecular interfaces. PMID- 11014595 TI - Amino acid-specific isotopic labeling and active site NMR studies of iron(II)- and iron(III)-superoxide dismutase from Escherichia coli. AB - We have developed and employed multiple amino acid-specific isotopic labeling schemes to obtain definitive assignments for active site 1H NMR resonances of iron(II)- and iron(III)-superoxide dismutase (Fe(II)SOD and Fe(III)SOD) from Escherichia coli. Despite the severe relaxivity of high-spin Fe(III), we have been able to assign resonances to ligand His' delta1 protons near 100 ppm, and beta and alpha protons collectively between 20 and 50 ppm, in Fe(III)SOD. In the reduced state, we have assigned all but 7 ligand protons, in most cases residue specifically. A pair of previously unreported broad resonances at 25.9 and 22.1 ppm has been conclusively assigned to the beta protons of Asp 156, superseding earlier assignments (Ming et al. (1994) Inorg. Chem., 33, 83-87). We have exploited higher temperatures to resolve previously unobserved ortho-like ligand His proton resonances, and specific isotopic labeling to distinguish between the possibilities of 82 and epsilon1 protons. These are the closest protein protons to Fe(II) and therefore they have the broadest (approximately 4,000 Hz) and most difficult to detect resonances. Our assignments permit interpretation of temperature dependences of chemical shifts, pH dependences and H/D exchange rates in terms of a hydrogen bond network and the Fe(II) electronic state. Interestingly, Fe(II)SOD's axial His ligand chemical shifts are similar to those of the axial His ligand of Rhodopseudomonas palustris cytochrome c' (Bertini et al. (1988) Inorg. Chem., 37, 4814-4821 ) suggesting that Fe(II)SOD's equatorial His2Asp- ligation is able to reproduce some of the electronic, and thus possibly chemical, properties of heme coordination for Fe2+. PMID- 11014596 TI - Radiation damping compensation of selective pulses in water-protein exchange spectroscopy. AB - The observation of nuclear Overhauser effects (NOEs) between bound water and biological macromolecules such as proteins and nucleic acids can be improved by inverting the water resonance selectively while compensating for radiation damping effects. The efficiency of inversion, the offset profiles, and the appearance of 2D NOE-NOESY spectra can be improved in comparison with earlier methods. PMID- 11014597 TI - Bridging the gap: A set of selective 1H-15N-correlations to link sequential neighbors of prolines. AB - Triple-resonance experiments are standard in the assignment of protein spectra. Conventional assignment strategies use 1H-15N-correlations as a starting point and therefore have problems when proline appears in the amino acid sequence, which lacks a signal in these correlations. Here we present a set of amino acid selective pulse sequences which provide the information to link the amino acid on either side of proline residues and thus complete the sequential assignment. The experiments yield amino acid type selective 1H-15N-correlations which contain signals from the amino protons of the residues either preceding or following proline in the amino acid sequence. These protons are correlated with their own nitrogen or with that of the proline. The new experiments are recorded as two dimensional experiments and their performance is demonstrated by application to a 115-residue protein domain. PMID- 11014598 TI - Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR studies. AB - Deuterium isotope labelling is important for NMR studies of large proteins and complexes. Many eukaryotic proteins are difficult to express in bacteria, but can be efficiently produced in the methylotrophic yeast Pichia pastoris. In order to facilitate NMR studies of the malaria parasite merozoite surface protein-1 (MSP1) complex and its interactions with antibodies, we have investigated production of the MSP1-19 protein in P. pastoris grown in deuterated media. The resulting deuteration patterns were analyzed by NMR and mass spectrometry. We have compared growth characteristics and levels of heterologous protein expression in cells adapted to growth in deuterated media (95% D2O), compared with expression in non adapted cells. We have also compared the relative deuteration levels and the distribution pattern of residual protiation in protein from cells grown either in 95% D2O medium with protiated methanol as carbon source, or in 95% D2O medium containing deuterated methanol. A high level of uniform Calpha deuteration was demonstrated, and the consequent reduction of backbone amide signal linewidths in [1H/15N]-correlation experiments was measured. Residual protiation at different positions in various amino acid residues. including the distribution of methyl isotopomers, was also investigated. The deuteration procedures examined here should facilitate economical expression of 2H/13C/15N-labelled protein samples for NMR studies of the structure and interactions of large proteins and protein complexes. PMID- 11014599 TI - 1H, 13C, and 15N backbone assignments of the ligand binding domain of TGFbeta type II receptor. PMID- 11014600 TI - NMR backbone assignments of the cold-regulated RNA-binding protein, rbpA1, in the cyanobacterium, anabaena variabilis M3. PMID- 11014601 TI - Sequence-specific NMR resonance assignments of the backbone atoms for the olfactory marker protein, OMP. PMID- 11014602 TI - Assignment of 1H, 13C and 15N signals of bovine adrenodoxin. PMID- 11014603 TI - Backbone 1H, 13C, and 15N resonance assignments of the anti-dansyl antibody Fv fragment. PMID- 11014604 TI - Backbone 1H, 15N, and 13C resonance assignments of inhibitor-2 -- a protein inhibitor of protein phosphatase-1. PMID- 11014605 TI - Isoflurane anaesthetic depth in goats monitored using the bispectral index of the electroencephalogram. AB - The bispectral index (BIS) of the electroencephalogram has recently been used to monitor the depth of anaesthesia in humans. The BIS is a dimensionless number that varies between 0 and 100. We hypothesized that the BIS could also be used to monitor depth of isoflurane anaesthesia in goats. Needle electrodes were placed over the frontal region of the scalp of goats and 5%, isoflurane was administered via a mask. The BIS number was determined at clinically relevant end-points. The BIS number did not change when the animals became recumbent (95 +/- 5 to 94 +/- 7, n = 15), but decreased to 65 +/- 13 and 64 +/- 15 when the corneal reflex and withdrawal response to a noxious stimulus, respectively, were lost (p < 0.001, n = 12). Direct laryngoscopy and intubation increased the BIS (56 +/- 7 to 83 +/- 11; p < 0.05, n = 10), as did a noxious pinch to the dew-claw (57 +/- 9; to 76 +/ 9; p < 0.05, n = 10). The spectral edge (frequency below which 95% of the total power resided) paralleled the change in BIS. We conclude that the depth of isoflurane anaesthesia in goats can be monitored using the BIS, although further work is needed to determine its sensitivity and specificity. PMID- 11014606 TI - Aeromonas hydrophila isolated from wild freshwater fish in Croatia. AB - Aeromonas hydrophila was recovered from fish living in lake Vrana on the Croatian island of Cres. The occurrence of the bacterium in the fish was assessed and related to gross signs of disease and findings at necropsy as a potential health hazard for fish. Isolated bacteria were subjected to morphological, physiological, biochemical and antibiotic susceptibility tests. A total of 26 A. hydrophila isolates were obtained. There was a clear seasonality, as no isolates were recovered in the summer months. Most of the isolates were sensitive to all the antimicrobials used in the study except novobiocin and penicillin G. Affected fish manifested haemorrhages over the skin, in the liver, kidney and swim bladder, spleen infarcts, fatty liver, ascitic fluid and swollen haemopoietic tissues. A. hydrophila does not appear to pose a major threat for the fish in the lake at present but under unfavourable and stressful conditions it could seriously compromise fish health. PMID- 11014607 TI - In vitro metabolism of fumonisin B1 by ruminal microflora. AB - Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium moniliforme and F. proliferatum. Little is known of its metabolic fate after oral ingestion in ruminants, but these animals are reported to be tolerant towards FB1. The metabolism of this mycotoxin was evaluated following incubation (1 microg/ml) in ruminal fluid for up to 72 h, in the presence or absence of alfalfa as a substrate for microbial growth, using a model rumen (sealed flask, anaerobic conditions, exclusion of light, gentle agitation, 39 degrees C). The decrease in FB1 concentration and the production of short-chain fatty acids were determined. FB1 had no effect on SCFA production. After 72 h incubation, FB1 depletion was 12% and 18% in samples with and without alfalfa, respectively. No hydrolysed metabolites (aminopolyols or aminopentol) were detected. These results indicate that FB1 is poorly metabolized in the rumen and suggest that such metabolism is not the cause of the tolerance to this toxin displayed by ruminants. PMID- 11014608 TI - Efficacy of copper oxide needles for the control of nematode parasites in dairy goats. AB - The spread of benzimidazole-resistant nematodes in dairy goat farms is of a great concern as probably more than 70% of the flocks are involved. As there are very few other anthelmintic options during the lactating period, we have evaluated the efficacy of copper oxide needles (CON, Copinox, Bayer, UK) in both experimental and natural infections in goats. The curative effect of CON (2-4 g) on existing worm burdens was assessed in goats experimentally infected with Teladorsagia circumcincta, Haemonchus contortus and Trichostrongyus colubriformis, compared to controls. The preventive effect of CON (4 g) on worm establishment was monitored for 2 months in animals experimentally infected with H. contortus and for 3 months in naturally infected animals on a farm exhibiting predominant infections with T. circumcincta and Oesophagostomum venulosum. In both experimental and natural conditions, the efficacy of CON was nil against Teladorsagia, Trichostrongylus and Oesophagostomum infections. In contrast, the efficacy of CON against Haemonchus was clearly established in reducing the worm burden (75%) as well as in lowering the egg output (37-95%) in relation to the establishment of new infections over several weeks. Copper oxide needles may represent an alternative to conventional anthelmintics in the control of Haemonchus infection in some goat farms. PMID- 11014609 TI - Characterization of an Indian bluetongue virus isolate by RT-PCR and restriction enzyme analysis of the VP-7 gene sequence. AB - The reverse transcription-polymerase chain reaction (RT-PCR) was standardized to amplify the VP-7 gene sequences of an Indian isolate of bluetongue virus serotype 23. Using two different sets of primers, a sequence of 1156 bp comprising the complete coding sequence of the VP-7 gene and its 770 bp internal sequence were amplified. The sensitivity of RT-PCR, using these two sets of primers individually was 40 pg and 4 pg, with the external and internal primers, respectively, whereas the nested PCR was 100-fold more sensitive than the single PCR with the external primers. Further, by restriction enzyme digestion of the 1156 bp amplicon, using CfoI, PstI and TaqI enzymes, the Indian isolate was found to be genetically different from isolates from the United States and Australia. RT-PCR and restriction enzyme digestion were applied to detect virus directly in blood samples taken from sheep suspected of bluetongue virus infection. PMID- 11014610 TI - Molecular typing of a BHV-4 (bovine herpesvirus 4) field isolate. AB - A new BHV-4 (bovine herpesvirus 4) isolated from a case of bovine interdigital dermatitis was characterized by PCR and restriction enzyme analysis. To determine whether the new isolate (PR/1) belonged to BHV-4, DNA from infected cells was specifically amplified by PCR. We used a set of primers spanning a large 2.571 kb conserved region of the BHV-4 genome, including the 3' end of ORF1 (homologous to the EBV BVRF1 gene), ORF2 (homologous to the EBV BXRF1 gene), ORF3 (TK gene) and ORF4 (gH gene) 5' end, respectively. The identity of the amplified product was confirmed by HindIII restriction enzyme digestion and Southern hybridization. No product was observed from the DNA of other bovine herpesviruses tested. The restriction patterns of the PR/ 1 genome compared to DN 599, MOVAR 33/63 and LVR BHV-4 reference strains showed two kinds of differences, either related or not related to the prDNA (polyrepetitive DNA). Taken together, these data show that PR/ 1 is a new BHV-4. We would consider that the present report provides a scheme of work for diagnosis and typing of BHV-4 isolates. PMID- 11014611 TI - Genomic and pathogenic studies on a glycoprotein E variant field isolate of bovine herpesvirus 1. AB - Glycoprotein E-negative (gE-) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability pf this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes. PMID- 11014612 TI - Behavioral despair in mice after prenatal stress. AB - Maternal stress during pregnancy produced behavioral alterations in both sexes with regard to sexual behavior, aggressive, maternal, lateralization and depression. In the present paper, sex differences for depression in mice was studied. No sex differences between female and male mice groups were observed either in swimming-induced immobility or in the open-field test (ambulation, rearing and boluses). Prenatal stress produced: 1) an increase of immobility time in female mice for swimming-induced immobility, but not in male mice; 2) an increase of ambulation in female mice for open-field test, but not in male mice; 3) there were no significant differences in rearing and boluses between stress and control groups either for female or male mice. Prenatal stress increases the risk of depression and locomotor activity in adult female mice. PMID- 11014613 TI - Prolactin and cyclosporine modulate adenosine transporters and adenosine A1 receptors in the rat brain. AB - The existence of adenosine A1 receptors and adenosine transporters in the central nervous system has been well demonstrated, although their possible modulation by hormones and/or exogenous drugs is poorly understood. To further analyze these modulatory mechanisms, the effects of prolactin and cyclosporine (CyA) on adenosine A1 receptors and transporters were analyzed in the central nervous system. For this purpose the number and affinity of adenosine A1 receptors were measured using the specific antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and the transporters with the high affinity ligand nitrobenzylthioinosine (NBTI). This procedure was carried out in hyperprolactinemic and control male rats treated with CyA or its vehicle for 8 days. As expected, pituitary grafting increased plasma prolactin levels (p<0.01). CyA treatment reduced but did not normalize (p<0.05) this parameter in hyperprolactinemic rats and did not modify circulating prolactin in control animals. Both hyperprolactinemia and CyA treatment reduced the number of adenosine transporters by 70% and by 40% the number of A1 receptors. The Kd for transporters was also reduced in all experimental groups. Hyperprolactinemia increased the affinity of A1 receptors (p<0.01) and CyA treatment did not further modify this parameter. These data demonstrated that prolactin and CyA influence adenosine transporters and A1 receptors at the central nervous system and suggest the existence of an interaction between prolactin and CyA may be operating to modulate these processes. PMID- 11014614 TI - Effects of IGF-I treatment on osteopenia in rats with advanced liver cirrhosis. AB - IGF-I is an anabolic hormone which has been reported to increase bone formation in several conditions of undernutrition. Advanced liver cirrhosis is associated with osteopenia and also with low serum levels of IGF-I. Previous results showed that low doses of IGF-I increase osteoblastic activity and decrease bone reabsorption in early liver cirrhosis. The aim of this study was to evaluate whether IGF-I-treatment also induces beneficial effect on osteopenia associated with advanced cirrhosis. Rats with ascitic cirrhosis were divided into two groups: group CI (n=10) which received saline and group CI+IGF (n=10) which were treated with IGF-I (2 microg/100 g bw x day, sc, during 21 days). Healthy controls which received saline were studied in parallel (CO n=10). On the 22nd day, the animals were sacrificed, and bone parameters were analyzed in femur. Posterior-anterior diameter was similar in all groups. No significant differences were observed in bone content of calcium, total proteins, collagen and hydroxyapatite in cirrhotic rats as compared with controls. However, CI rats showed significant reductions in total bone density (-13.5%, p<0.001) assessed by densitometry and radiological study. In CI+IGF rat bone density (assessed by densitometry) improved significantly as compared with CI animals. In summary, osteopenia characterized by loss of bone mass and preserved bone composition was found in rats with advanced cirrhosis induced by CCl4 and phenobarbital in drinking water. This bone disorder is partially restored by treatment with low doses of IGF-I during only three weeks. Thus, IGF-I could be considered as a possible therapy for osteopenia associated with advanced liver cirrhosis. PMID- 11014615 TI - Retinoid X receptor mRNA expression in human pituitary gland. AB - Retinoic acid is involved in important physiological processes such as the regulation of growth and differentiation of several tissues, including the pituitary gland. These biological effects are mediated by their binding to two specific intracellular receptors termed retinoic acid and retinoid X receptors, RARs, RXR, respectively). RAR or RXR mRNA expression has been demonstrated in several tissues, but little information is available about its presence in the human pituitary gland. In this report, we demonstrate alphaRXR mRNA expression using the reverse transcription coupled to polymerase chain reaction (RT-PCR) in the human pituitary gland. These results suggest the possibility that RXR may regulate the human pituitary gene expression and hormone secretion. PMID- 11014616 TI - Growth hormone response to long-term GH-RH administration in lambs. AB - The pattern of long-term GHRH administration capable of stimulating GH release without depleting pituitary GH content has been investigated using two experimental approaches. In experiment 1, recently weaned male lambs were treated for 3 weeks as follows: Group A) control; B) subcutaneous (sc) continuous infusion of GHRH (1200 mg/day) using a slow release pellet; C) the same as B plus 1 daily sc injection of long acting somatostatin (SS) (octreotide, 20 mg) ; D) 3 daily sc GHRH (250 mg) injections ; E) 2 daily sc injections of GHRH (250 mg) and 2 of natural SS (250 mg). In experiment 2, recently weaned male lambs were continuously GHRH-treated using sc osmotic minipumps (900 mg/day) alone or combined with a daily sc injection of octreotide (20 mg) for 4 weeks. Basal plasma GH levels were increased after chronic pulsatile GHRH treatment but not after any kind of continuous GHRH administration. This increment was maintained during the 3 weeks of experimentation and appeared accompanied by a pituitary GH content similar to controls. A marked GH response to the iv GHRH challenge was observed in controls and in lambs receiving both types of continuous sc GHRH infusions, whereas pulsatile sc GHRH-treated animals did not respond to the iv GHRH challenge in the first and second weeks of the study but did so in the third week of treatment. These data demonstrate that long-term pulsatile GHRH administration is capable of stimulating GH release in growing male lambs, without producing pituitary desensitization. PMID- 11014617 TI - Nutrient toxicity in pancreatic beta-cell dysfunction. AB - Nutrients, such as glucose and fatty acids, have a dual effect on pancreatic beta cell function. Acute administration of high glucose concentrations to pancreatic beta-cells stimulates insulin secretion. In addition, short term exposure of this cell type to dietary fatty acids potentiates glucose-induced insulin release. On the other hand, long-term exposure to these nutrients causes impaired insulin secretion, characterized by elevated exocytosis at low concentrations of glucose and no response when glucose increases in the extracellular medium. In addition, other phenotypic changes are observed in these conditions. One major step in linking these phenotypic changes to the diabetic pathology has been the recognition of both glucose and fatty acids as key modulators of beta-cell gene expression. This could explain the adaptative response of the cell to sustained nutrient concentration. Once this phase is exhausted, the beta-cell becomes progressively unresponsive to glucose and this alteration is accompanied by the irreversible induction of apoptotic programs. The aim of this review is to present actual data concerning the development of the toxicity to the main nutrients glucose and fatty acids in the pancreatic beta-cell and to find a possible link to the development of type 2 diabetes. PMID- 11014618 TI - Energy expenditure during leisure time and body mass index in Spain. PMID- 11014619 TI - Management of cytomegalovirus retinitis in AIDS. PMID- 11014620 TI - Clinical manifestations and molecular epidemiology of late recurrent candidemia, and implications for management. AB - The aim of this study was to define the epidemiology and clinical manifestations of late recurrent candidemia. For this purpose, late recurrent candidemia was defined as an episode of candidemia occurring at least 1 month after the apparent complete resolution of an infectious episode caused by the same Candida sp. A total of five patients with recurrent candidemia were investigated. For all patients, isolates from the initial and recurrent episodes of candidemia were available for in vitro susceptibility testing and genetic characterization by DNA based techniques. The results revealed the following salient features: prolonged duration between the initial and recurrent episodes (range, 1-8 months); recurrence of candidemia despite anti-fungal therapy; importance of retained intravascular catheters, neutropenia, and corticosteroids as factors predisposing to recurrence; high morbidity and mortality; no emergence of antifungal drug resistance between the initial and recurrent episodes; and relapse of infection due to the original infecting strain, rather than reinfection with a new strain. These findings raise several issues about the management and follow-up of patients with candidemia, which require assessment in future studies. PMID- 11014621 TI - Determinants for the development of oropharyngeal colonization or infection by fluconazole-resistant Candida strains in HIV-infected patients. AB - A point prevalence study to document oral yeast carriage was undertaken. Risk factors for the development of oropharyngeal colonization or infection by fluconazole-resistant Candida strains in HIV-infected patients were investigated with a case-control design. Cases included all patients with fluconazole resistant strains (MIC> or =64 microg/ml), and controls were those with susceptible (MIC< or =8 microg/ml) or susceptible-dependent-upon-dose (MIC 16-32 microg/ml) strains. One hundred sixty-eight Candida strains were isolated from 153 (88%) patients, 28 (16%) of whom had oropharyngeal candidiasis. Overall, 19 (12%) of the patients harbored at least one resistant organism (MIC > or = 64 microg/ml). Among patients with resistant strains, tuberculosis (P<0.001), esophageal candidiasis (P = 0.001), clinical thrush (P<0.001), and a CD4 + cell count < 200/mm3 (P = 0.03) were more frequent. These patients had also been treated more commonly with antituberculous drugs (adjusted odds ratio [OR] 6.13; 95% confidence interval [CI] 2.11-17.80), ciprofloxacin (OR 6.0; 95% CI 1.23 29.26), fluconazole (OR 4.59; 95% CI 1.55-13.52), and steroids (OR 4.13; 95% CI 1.11-15.39). Multivariate analysis showed that the determinants for fluconazole resistance were therapy with antituberculous drugs (OR 3.61; 95% CI 1.08-12.07; P=0.03) and one of the following: previous tuberculosis (OR 3.53; 95% CI 1.08 14.57; P=0.03) or fluconazole exposure (OR 3.41; 95% CI 1.10-10.54). Findings from this study indicate that treatment with antituberculous drugs, previous tuberculosis, and fluconazole exposure are the strongest determinants for development of oropharyngeal colonization or infection by fluconazole-resistant Candida strains in HIV-infected patients. PMID- 11014622 TI - Nosocomial candidemia in non-neutropenic patients at an Italian tertiary care hospital. AB - In a retrospective study conducted in an Italian tertiary care hospital, the incidence of nosocomial candidemia was evaluated together with causative pathogens, treatment, and risk factors for death. Over a 6-year period (1992 1997), a total of 189 episodes of candidemia occurred in 189 patients (mean age 58+/-19 years), accounting for an average incidence of 1.14 episodes per 10,000 patient-days per year. The most common reasons for hospitalization were solid neoplasia (21%), trauma (17%), abdominal diseases requiring surgery (13%), and cardiovascular diseases (13%). No patient was neutropenic within 3 weeks prior to the onset of candidemia. One hundred thirty patients were hospitalized in intensive care units, 47 patients in surgical wards, and 12 patients in medical wards. Candida albicans was the most frequently isolated pathogen, accounting for 54% of fungal isolates, followed by Candida parapsilosis (23%), Candida glabrata (7%), Candida tropicalis (5%), Candida pelliculosa (4%), Candida lusitaniae (1%), Candida humicula (1%), and other non-albicans Candida spp. (5%). Seventy-six (41%) patients received adequate antifungal therapy. Seventy-one (58%) of the 123 evaluable patients with central venous catheters underwent line removal; 51 of them had catheter-related candidemia. The 30-day crude mortality rate was 45%. Older age, hospitalization in an intensive care unit, a longer duration of candidemia, retention of central lines, and inadequate antifungal therapy were significantly associated with poor outcome. In the present study, nosocomial candidemia was a frequent and relatively underestimated illness. Adequate antifungal therapy and central line removal independently reduced the high mortality of the disease. PMID- 11014623 TI - Isolation of Kingella kingae from synovial fluids using four commercial blood culture bottles. AB - According to the literature, Kingella kingae may be an underdiagnosed cause of joint and bone infections in children. The use of the Bactec blood culture system for culture of joint fluids has dramatically improved the isolation of this fastidious bacterium. The aim of this study was to test the recovery rate and detection time of four commercial blood culture systems: three different BacT/Alert (Organon Teknika, USA) bottles and one Bactec (Becton Dickinson Microbiology Systems, USA) bottle, all inoculated with Kingella kingae strains mixed with pooled synovial fluids. For each strain the same inoculum and volume of synovial fluid was distributed into each of the four bottles. All 24 strains tested grew in the BacT/Alert Aerobic (100%) and the BacT/Alert Pedi-BacT (100%) bottles. Twenty-one strains grew in the BacT/Alert FAN aerobic (88%) bottle, and 15 strains grew in the Bactec Plus Aerobic F (63%) bottle, in both systems within 12 days (P<0.01). The Kingella kingae strains were first detected in the BacT/Alert Pedi-BacT bottles (P<0.001). The results were reproducible. The BacT/Alert blood culture bottles were superior to previously described blood culture systems in isolating Kingella kingae from synovial fluid, even with small inoculums and small volumes of synovial fluid. PMID- 11014624 TI - In vitro zones of inhibition of coated vascular catheters predict efficacy in preventing catheter infection with Staphylococcus aureus in vivo. AB - This report summarizes data from 35 rabbit model experiments investigating the relationship between in vitro anti-infective catheter coating zones of inhibition and in vivo efficacy. The rabbit model studies involving 15 anti-infective coatings demonstrate an inverse correlation between the sizes of zones of inhibition of Staphylococcus aureus and both the quantity of Staphylococcus aureus removed from the catheter and the risk of a purulent infection. The review of seven previously published clinical trials reveals that the use of anti infective coated catheters, efficacious in the rabbit model, was associated with a higher success rate than the use of uncoated catheters in preventing both Staphylococcus aureus catheter colonization (odds ratio: 1.28; 95% confidence interval: 0.84-1.93) and Staphylococcus aureus catheter-related bloodstream infection (odds ratio: 3.07; 95% confidence interval: 0.98-9.60) in humans. These findings strongly suggest a correlation between zones of inhibition and in vivo efficacy. In vitro zones of inhibition may serve as a useful screening test for evaluating new anti-infective coatings. PMID- 11014625 TI - Evaluation of the Vitek 2 system for susceptibility testing of Streptococcus pneumoniae isolates. AB - Vitek 2 (bioMerieux, France) is a new commercial system that allows rapid identification and rapid determination of the minimum inhibitory concentration (MIC) of Streptococcus pneumoniae by monitoring the growth kinetics of the organisms in microwells. The accuracy of the Vitek 2 system in susceptibility testing was evaluated by determining the MICs of 50 penicillin-susceptible and 150 intermediate or penicillin-resistant Streptococcus pneumoniae isolates and comparing the results with those obtained using the agar dilution method. The essential agreement between the Vitek 2 system and the reference method was 91% for penicillin, 93% for cefotaxime and ceftriaxone, and more than 94% for amoxicillin, erythromycin, ofloxacin, co-trimoxazole, tetracycline, and imipenem. One very major error (1.1%) and one major error (0.9%) were obtained for tetracycline. The minor error rate for penicillin of 19.3% was mainly due to intermediate category isolates (n = 29) being identified as resistant and susceptible isolates (n = 6) being identified as intermediate by the commercial system. The minor error rates for amoxicillin, cefotaxime, ceftriaxone, imipenem, and ofloxacin were 25.4%, 25.4%, 29.4%, 19.2%, and 31.5%, respectively. Vancomycin, tetracycline, co-trimoxazole, and erythromycin showed minor error rates of 0-6.1%. In conclusion, Vitek 2 shows good agreement with the reference method, as demonstrated by the low numbers of major errors, but it has a tendency to overestimate MICs, resulting in minor errors. PMID- 11014626 TI - Response of lymphocyte subsets in patients under treatment for tuberculosis. AB - Thirty-six immunocompetent patients with tuberculosis underwent six sequential measurements of blood lymphocyte subsets after the onset of therapy. Two different responses were observed, depending on each patient's initial lymphocyte count. Patients with initial lymphocytopaenia showed a marked increase in all lymphocyte subpopulations shortly after the initiation of treatment, whereas those without lymphocytopaenia showed a decrease in the cell counts during the initial 2 months, followed by a slow increase during the following 4 months. The lymphocytes as a whole and all lymphocyte subsets showed remarkably parallel curves in each group. These results strongly suggest that the lymphocytic responses to tuberculosis involve all lymphocyte subsets, not only the T or CD4+ lymphocytes as previously thought. PMID- 11014627 TI - Molecular typing of bacteria directly from cerebrospinal fluid. AB - Using Streptococcus pneumoniae as an example, the ability of multilocus sequence typing (MLST) to characterise isolates directly from cerebrospinal fluid (CSF) was investigated. A nested multiplex polymerase chain reaction method that amplifies the seven housekeeping gene fragments used for pneumococcal MLST was applied to 30 CSF samples from suspected cases of bacterial meningitis. The fragments were amplified from all 14 samples from which Streptococcus pneumoniae was cultured, and, after direct sequencing, the allelic profiles obtained from ten of the samples corresponded to those of clones previously associated with invasive pneumococcal disease. MLST could also predict the penicillin susceptibility and serotype of the CSF isolates. PMID- 11014628 TI - Detection of Isospora belli by polymerase chain reaction using primers based on small-subunit ribosomal RNA sequences. AB - The aim of the present study was to use small-subunit (SSU)-rRNA sequences of Isospora belli to design specific primer pairs and a hybridization probe for the detection of Isospora belli in human samples by PCR and Southern blot hybridization. PCR amplification with the primer pairs produced correct DNA fragments with target DNA from samples of Isospora belli-infected patients and from cloned SSU-rRNA of Isospora belli. The nature of the PCR products was confirmed by Southern blot hybridization. No amplification was seen with template DNA extracted from other parasites. Although Isospora belli infections can be easily diagnosed using light microscopy, molecular-based techniques may prove useful as an additional diagnostic tool. PMID- 11014629 TI - In vitro activity of ABT-773 against anaerobic bacteria. AB - The purpose of the study reported here was to determine the in vitro activity of ABT-773 compared with that of other antimicrobial agents against anaerobic bacteria. The activity of ABT-773 was investigated against 354 clinical isolates of anaerobic bacteria by the agar dilution method and was compared with that of azithromycin, clarithromycin, roxithromycin, erythromycin, cefoxitin, imipenem, clindamycin and metronidazole. ABT-773 and imipenem were the most active antimicrobial agents tested. PMID- 11014630 TI - Prosthetic valve endocarditis caused by Mycoplasma hominis. AB - Mycoplasma hominis endocarditis is extremely uncommon and difficult to diagnose. Atypical growth characteristics in routine bacterial culture and an inability to demonstrate the organism using Gram staining can lead to a delayed diagnosis of Mycoplasma hominis infections, and the organism is often missed. This report describes a patient with Mycoplasma hominis prosthetic valve endocarditis. The microorganism was recovered from the mitral prosthesis but was missed in blood cultures. This finding suggests that Mycoplasma hominis should be considered in the differential diagnosis of culture-negative endocarditis. PMID- 11014631 TI - Otitis externa due to Trichoderma longibrachiatum. PMID- 11014632 TI - Mixed community-acquired fungal infection in an apparently healthy patient. PMID- 11014633 TI - Detection of Pneumocystis carinii DNA in patients with chronic lung diseases. PMID- 11014635 TI - Molecular analyses of possible mechanisms coding for low-level mupirocin resistance in clinical Staphylococcus aureus isolates. PMID- 11014634 TI - In vitro activity of a new liposomal nystatin formulation against opportunistic fungal pathogens. PMID- 11014636 TI - Oral leishmaniasis in an HIV-infected patient. PMID- 11014637 TI - Understanding rejection of xenografts. PMID- 11014638 TI - Epithelial re-growth is associated with inhibition of obliterative airway disease in orthotopic tracheal allografts in non-immunosuppressed rats. AB - BACKGROUND: Because epithelial cells are targets of alloimmune injury leading ultimately to airway obliteration, we tested whether epithelial re-growth could prevent obliterative airway disease (OAD) in orthotopic tracheal allografts. METHODS: Brown Norway tracheal segments were orthotopically transplanted into nonimmunosuppressed Lewis rats. Allografts were removed on days 2-10 (n=13), 30 (n=4), and 60 (n=5) for histology, computerized morphometry (obliteration), and immunohistochemical detection of mononuclear cells, smooth muscle alpha-actin, and tissue phenotype. Normal tracheas, host tracheas, and heterotopically transplanted allografts served as controls. RESULTS: Orthotopic allografts removed on days 2-10 exhibited epithelial damage and re-growth and mononuclear cell infiltration. On days 30 and 60, partially ciliated cuboidal or attenuated epithelium completely covered the lumen. Although mononuclear cells declined, numerous T cells with a high CD4/CD8 ratio were found in the epithelium till day 60. Orthotopic allograft epithelium expressed donor phenotype on day 7, but recipient phenotype on days 30 and 60. Despite subepithelial alpha-actin positive myofibroblast proliferation, obliteration did not progress from day 7 to 30 and 60 (35, 30, and 33%, respectively). Although more than in normal or host tracheas, the obliteration in orthotopic allografts on days 30 and 60 was significantly less (P<0.001) than in heterotopic allografts. CONCLUSIONS: We describe, for the first time, longterm patency of fully histoincompatible orthotopic tracheal allografts in nonimmunosuppressed rats. Despite acute alloimmune injury and induction of myofibroblast proliferation, epithelial re growth from the host limited the progression of OAD, thus emphasizing the role of epithelium in the control of airway obliteration. PMID- 11014639 TI - Recombinant adenoviral mediated CD39 gene transfer prolongs cardiac xenograft survival. AB - BACKGROUND: Extracellular ATP and ADP may be important mediators of vascular inflammation and thrombosis. Nucleoside triphosphate diphosphohydrolase (NTPDase or CD39) is a vascular ectoenzyme that hydrolyses ATP and ADP; however, this activity is lost during reperfusion injury. We show that the supplementation of NTPDase activity within xenograft vasculature using CD39 recombinant adenoviruses (AdCD39) has protective effects in vivo. METHODS: Recombinant adenoviruses containing human CD39 or beta-galactosidase (Adbeta-gal) encoding genes were constructed. Hartley guinea pig coronary arteries were perfused ex vivo with University of Wisconsin solution containing 10(9) plaque-forming units of the recombinant adenovirus. Infected grafts were then implanted in the abdomen of complement depleted Lewis rats. RESULTS: NTPDase activities decreased in all grafts within the first 24 hr and subsequently recovered only in those hearts infected with AdCD39. Immunohistological examination of AdCD39-infected grafts confirmed successful CD39 gene transfer into the endocardium and macrovasculature. Expression of CD39 modestly prolonged graft survival (90.2+/ 5.4 hr, mean+/-SD, n=5) when compared with Adbeta-gal-infected grafts (67.4+/-5.4 hr, P<0.005) and perfusion controls (66.4+/-5.2 hr; P<0.005). CONCLUSIONS: Recombinant adenoviral infection can induce expression of CD39 within cardiac xenografts and provide survival benefits in vivo. Our data show that ex vivo infection by recombinant adenovirus vectors can result in vascular expression of a potential therapeutic agent. PMID- 11014640 TI - Successful prevention of autoimmune disease by transplantation of adequate number of fully allogeneic hematopoietic stem cells. AB - BACKGROUND: We have previously shown successful engraftment of allogeneic hematopoietic stem cells (HSCs) when transplanted across the major histocompatibility antigen barriers if transplanted along with a preparation of facilitator cells (osteoblasts). We have investigated whether or not fully allogeneic HSCs from healthy mouse donors prevent the development of autoimmunities in the autoimmune-prone W/B F1 mice. METHODS: W/B F1 is a strain of mice that spontaneously develop autoimmunities, a coronary vascular disease, thrombocytopenia, and systemic lupus-like syndrome. The 6- to 8-week-old (before the onset of the disease) W/B F1 mice have been transplanted with either a preparation of HSCs alone, or along with facilitator cells from MHC-incompatible autoimmune-resistant BALB/c mice, then followed to determine longterm survival and whether or not they developed signs of the autoimmune disease. RESULTS: The number of the transplanted HSCs acts as the determining factor in achieving successful and durable engraftment. Survival of the W/B F1 mice significantly improved by transplantation of increasing numbers of HSCs, either alone or along with facilitator cells. When W/B F1 mice were transplanted with 2-5 million HSCs, more than 1-year survival was 100%, all the transplanted mice were fully engrafted with allogeneic HSCs, and were free of signs of the autoimmune disease. Histological sections of the hearts, lungs, and kidneys of the transplanted mice showed absence of the autoimmune-associated pathology. CONCLUSIONS: We thus report herein the successful prevention of autoimmune disease by transplantation of a sufficiently large number of purified fully allogeneic HSCs in W/B F1 mice. PMID- 11014641 TI - Increased glomerular deposits of von Willebrand factor in chronic, but not acute, rejection of primate renal allografts. AB - BACKGROUND: In our previously described primate renal allograft model, T cell ablation leads to long-term graft survival. The role of endothelial cell alteration in chronic rejection was examined in our model. METHODS: Renal transplants were performed in rhesus monkeys using a T cell- depleting immunotoxin, FN18-CRM9. Sections from 10 rejected kidneys (5 acute and 7 chronic rejection) were examined after immunohistochemical staining for expression of endothelium-related proteins [von Willebrand factor (vWF), CD62P, and CD31], fibrinogen, and a macrophage marker (CD68). Glomerular staining for each antigen was graded on a semiquantitative scale. RESULTS: Intense staining for vWF was consistently observed in glomerular endothelium, subendothelium, and mesangium in all kidneys removed due to chronic rejection. vWF staining was weak in kidneys showing acute rejection. The difference in glomerular staining was statistically significant. Staining for vWF in extraglomerular vessels was nearly identical in kidneys showing acute and chronic rejection. Expression of CD62P was increased in extraglomerular vessels in allografts with chronic rejection, but the glomeruli showed little or no staining. There was no significant difference in the glomerular staining for CD62P or CD31 in organs showing acute and chronic rejection. Fibrinogen staining of glomerular mesangium was seen in kidneys with chronic rejection. Macrophages (CD68+) infiltrating glomeruli were more numerous in kidneys showing chronic rejection. CONCLUSION: Increased glomerular deposition of vWF in renal allografts showing chronic rejection, without increased staining for CD62P or CD31, suggests increased constitutive secretion of vWF from endothelial cells as a component of the mechanism of chronic rejection in our model. PMID- 11014642 TI - Plasmapheresis and intravenous immune globulin provides effective rescue therapy for refractory humoral rejection and allows kidneys to be successfully transplanted into cross-match-positive recipients. AB - BACKGROUND: Hyperacute rejection (HAR) and acute humoral rejection (AHR) remain recalcitrant conditions without effective treatments, and usually result in graft loss. Plasmapheresis (PP) has been shown to remove HLA- specific antibody (Ab) in many different clinical settings. Intravenous gamma globulin (IVIG) has been used to suppress alloantibody and modulate immune responses. Our hypothesis was that a combination of PP and IVIG could effectively and durably remove donor-specific, anti-HLA antibody (Ab), rescuing patients with established AHR and preemptively desensitizing recipients who had positive crossmatches with a potential live donor. METHODS: The study patients consisted of seven live donor kidney transplant recipients who experienced AHR and had donor-specific Ab (DSA) for one or more mismatched donor HLA antigens. The patients segregated into two groups: three patients were treated for established AHR (rescue group) and four cross match-positive patients received therapy before transplantation (preemptive group). RESULTS: Using PP/IVIG we have successfully reversed established AHR in three patients. Four patients who were cross-match-positive (3 by flow cytometry and 1 by cytotoxic assay) and had DSA before treatment underwent successful renal transplantation utilizing their live donor. The overall mean creatinine for both treatment groups is 1.4+/-0.8 with a mean follow up of 58+/-40 weeks (range 17 116 weeks). CONCLUSIONS: In this study, we present seven patients for whom the combined therapies of PP/IVIG were successful in reversing AHR mediated by Ab specific for donor HLA antigens. Furthermore, this protocol shows promise for eliminating DSA preemptively among patients with low-titer positive antihuman globulin-enhanced, complement-dependent cytotoxicity (AHG-CDC) cross-matches, allowing the successful transplantation of these patients using a live donor without any cases of HAR. PMID- 11014643 TI - CD8+ T cells are capable of rejecting pancreatic islet xenografts. AB - BACKGROUND: In this study, the capacity of CD8+ T cells to act as a potential effector mechanism in pancreatic xenograft rejection was examined. METHODS: The fate of pancreatic islet xenografts was studied in mice deficient in MHC class II molecules and CD4+ T cells. Fetal pig pancreas (FPP) or Wistar rat islets (RI) were transplanted into nondiabetic or streptozotocin-induced diabetic I-A knock out (CII K/O) mice. RESULTS: CII K/O mice were capable of rejecting both RI and FPP grafts. RI graft survival was not prolonged compared with wild type C57BL/6 controls. However, FPP grafts did survive longer in CII K/O recipients than in C57BL/J6 mice. Both RI and FPP graft rejection were CD8+ T-cell phenomena in CII K/O mice, as anti-CD8 monoclonal antibody prolonged graft survival, there were increased CD8+ T cells in the grafts and spleens of CII K/O recipients, and cell mediated cytotoxicity was a CD8+ T-cell phenomenon associated with activation of the perforin/granzyme B system. By contrast, RI and FPP graft rejection was a CD4+ T cell-dependent phenomenon in wild type C57BL/6 mice with graft survival prolonged by anti-CD4 monoclonal antibody. There were increased numbers of CD4+ T cells, and cell-mediated cytotoxicity was a CD4+ T-cell phenomenon associated with activation of the Fas/FasL lytic pathway. CONCLUSIONS: The results demonstrate that, in the absence of CD4+ T cells, CD8+ T cells were capable of rejecting both rat and pig pancreatic islet xenografts. PMID- 11014644 TI - Mixed chimerism, heart, and skin allograft tolerance in cyclophosphamide-induced tolerance. AB - We elucidated the possible role of chimerism in skin and heart allograft tolerance using cyclophosphamide (CP)-induced tolerance. When C3H (H-2k; Thy1.2, Mls-1b) mice were i.v. primed with 1x10(8) spleen cells (SC) from H-2 matched AKR (H-2k; Thy1.1, Mls-1a) mice and then treated i.p. with 200 mg/kg of CP, the survivals of both AKR skin grafts and heart grafts (HG) were permanently prolonged in a tolerogen-specific fashion. After this treatment, a minimal degree of mixed chimerism, the clonal destruction of Mls-1a-reactive CD4+Vbeta6+ T cells in the periphery, and the clonal deletion of Vbeta6+ thymocytes were all observed. When AKR SC and 100 mg/kg CP were used for conditioning, the AKR HG were permanently accepted, but the survival of the AKR skin grafts was only mildly prolonged. The clonal destruction of CD4+Vbeta6+ T cells in the periphery and the intrathymic clonal deletion of Vbeta6+ thymocytes were induced in both the SC and the 100 mg/kg CP-treated C3H mice. A minimal degree of mixed chimerism was detectable at 4 and 12 weeks after AKR SC and 100 mg/kg CP treatment, and still did not disappear at 40 weeks. The degree of mixed chimerism induced with SC and 100 mg/kg CP was significantly lower than that with SC and 200 mg/kg CP during the observation. No posttransplant cardiac allograft vasculopathy (CAV) was observed to develop, while both the Th1 type (interferon-gamma) and Th2 type (interleukin-4 and -10) cytokine expressions decreased in the AKR HG of the tolerant C3H mice treated with both AKR SC plus 200 mg/kg CP, and AKR SC plus 100 mg/kg CP. A second set of skin grafts from donor AKR mice survived for more than 100 days in a tolerogen-specific fashion in all C3H mice treated with AKR SC and 200 mg/kg CP and also accepted the AKR HG for over 200 days, while 80% of the C3H mice treated with AKR SC and 100 mg/kg CP and accepted the AKR HG for more than 200 days. These results strongly suggested the following conclusions: 1) the degree of chimerism can strongly influence the induction of skin and heart allograft tolerance, 2) posttransplant CAV does not develop in the donor HG maintained by chimerism-based CP-induced tolerance, 3) the mRNA expression of both Th1 and Th2 type cytokine decreased in the donor HG maintained by chimerism based CP-induced tolerance, and 4) the induction of skin allograft tolerance is more difficult than the prevention of posttransplant CAV. PMID- 11014645 TI - Alpha-gal-independent dual recognition and activation of xenogeneic endothelial cells and human naive natural killer cells. AB - BACKGROUND: Interaction between vascularized xenograft and host immune system is thought to occur via Galactose alpha (1,3) Galactose (Gala 1,3 gal) structures decorating the xenograft. METHODS: We raised anti-Gala 1,3 gal-BSA polyclonal antibodies in baboons and investigated effect(s) of these antibodies as well as soluble Gala 1,3 gal-BSA on human naive natural killer (NK) cell interactions with porcine aortic endothelial cells. RESULTS: We demonstrate that human naive (unstimulated) NK cells recognize xenogeneic endothelial cells under conditions where binding to the Gala 1,3 gal structures is minimized by the presence of blocking anti-Gala 1,3 gal IgG or soluble Gala 1-3 gal and in the absence of xenoreactive natural antibodies and complement. After xenogeneic encounter both endothelial cells and human NK cells are activated. Endothelial cell activation is rapid and is manifested initially by an intraendothelial calcium transient and subsequently by expression of P-selectin and vascular endothelial cell adhesion molecule-1 on the xenoendothelium surface. NK cell activation is manifested by increased expression of perforin and increased cytotoxicity towards the xenoendothelium. Neither recognition nor activation of the xenoendothelium was affected by the introduction of either anti-Gala 1,3 gal IgG or soluble Gala 1-3 gal. CONCLUSION: Our data provide evidence that innate immune cells, such as NK cells, recognize and activate xenoendothelial cells independently of Gala 1-3 gal structures and raise the possibility of novel interactive sites on both human naive NK cells and discordant xenogeneic endothelium. PMID- 11014646 TI - Interleukin-13 protects endothelial cells from apoptosis and activation: association with the protective genes A20 and A1. AB - BACKGROUND: Chronic rejection is the major obstacle to long-term survival of allografts and is associated with graft endothelial cell activation and apoptosis. Recent reports have found an association between graft survival, presence of Th2 cytokines, and expression by endothelial cells of cytoplasmic "protective" molecules that prevent apoptosis and down-regulate the inflammatory process. METHODS: Cultured human umbilical vein endothelial cells (HUVEC) were used. Apoptotic cells were detected by staining with FITC-annexinV followed by flow cytometry. Expression of vascular cell adhesion molecule-1, E-selectin, and intercellular adhesion molecule-1 were also measured by flow cytometry. Transcripts were detected by reverse transcription-PCR and quantitation was achieved by co-amplification of competing, internal standard RNA. RESULTS: We demonstrate that exposure of HUVEC to interleukin (IL)-13 for 72 hr afforded partial protection from apoptosis induced by tumor necrosis factor alpha/cycloheximide or serum starvation. Pretreatment with IL-13 also modulated induction of E-selectin after acute exposure to tumor necrosis factor-alpha or IL 1alpha. Protection was associated with transcription of the genes A1 and A20. Prolonged treatment with IL-13 had minimal proinflammatory effects and did not induce expression of E-selectin or vascular cell adhesion molecule-1 or increase intercellular adhesion molecule-1 above basal levels. CONCLUSIONS: Our data provide a possible explanation for the observed association between Th2 cytokines and expression of protective genes in the endothelium of long-surviving allografts and xenografts. PMID- 11014647 TI - Specific depletion of preformed IgM natural antibodies by administration of anti mu monoclonal antibody suppresses hyperacute rejection of pig to baboon renal xenografts. AB - BACKGROUND: The elimination of circulating anti-porcine preformed antibodies is crucial for avoiding hyperacute vascular rejection (HAVR) of primarily vascularized xenograft in discordant pig to baboon model. Previously described methods used for eliminating natural antibodies, however, constantly removed both anti-porcine IgM and IgG antibodies, as well as often complement proteins. To study specifically the role of preformed anti-porcine IgM antibodies, a specific anti-IgM monoclonal antibody (mAb) has been designed and evaluated in vivo. METHODS: Iterative injections of anti-IgM mAb (LO-BM2) at high dose (20 mg/kg) depleted to undetectable level the circulating IgM and therefore anti-porcine IgM antibodies but did not change the concentration of anti-pig IgG antibodies. The serum concentration of IgM and IgG antibodies was assessed by ELISA and the level of anti-pig natural IgM and IgG antibodies by flow cytometry (FC). Anti-rat sensitization was assessed by specific ELISA as well as the serum concentration of LO-BM2. RESULTS: Iterative injections of LO-BM2 allowed to specifically eliminate the anti-porcine IgM antibodies to undetectable levels at ELISA. Despite a normal serum level of anti-porcine IgG and complement proteins, HAVR was avoided. Without immunosuppression, the specific elimination of preformed anti-porcine IgM prolonged the survival of a renal xenograft in baboon up to 6 days, whereas without IgM antibody elimination, the renal xenografts were hyperacutely rejected within hours. The lost of activity of LO-BM2 after 10 days was concomitant to an IgM and IgG antibody rebound, which caused an acute vascular rejection of the xenograft. CONCLUSION: Specific elimination of natural anti-porcine IgM antibodies allows to avoid HAVR of a pig to baboon renal xenograft, whereas anti-porcine IgG antibodies and complement proteins were present in the serum. This result confirms previous in vitro reports and demonstrates for the first time in vivo that preformed IgM antibodies alone are responsible for HAVR, while preformed anti-porcine IgG antibodies are unable alone to cause HAVR. Anti-IgM therapy appears as an important tool to transiently but completely eliminates xeno-IgM antibodies in vivo. PMID- 11014648 TI - Local production of anti-CD4 antibody by transgenic allogeneic grafts affords partial protection. AB - BACKGROUND: Immunosuppressive drugs and anti-lymphocyte antibody are used clinically to suppress cellular rejection responses. However, these systemic regimens often led to general immunodeficiency and thus increased susceptibility to opportunistic infection and neoplasia. Immunosuppressive molecules delivered locally may be a way of inhibiting rejection responses, whereas systemic immunity is preserved. To achieve protective local immunosuppression, we produced a graft secreting its own immunomodulator, by deriving transgenic mice expressing a chimeric anti-CD4 antibody (GK2c) in the pancreas. METHODS AND RESULTS: Transgenic mice in bml genetic background expressing a modified anti-mouse CD4 antibody (GK2c) under two promoters have been produced. Tissue expression of GK2c was detected by immunoperoxidase staining. Under the cytomegalovirus promoter, there was abundant GK2c expression in pancreatic exocrine tissue. Under the rat preproinsulin II promoter, there was abundant GK2c expression in pancreatic endocrine tissue only. High-expression transgenic lines had 10-100 microg/ml GK2c in blood plasma. By flow cytometry, these transgenic mice were devoid of CD4+ cells in their peripheral lymphoid organs. To test transgenic mice as donors, fetal pancreata from transgenic mice were grafted into fully allogeneic CBA mice under the kidney capsule, transgenic grafts had prolonged survival compared with control non-transgenic grafts. Furthermore, GK2c transgenic grafts had reduced infiltration with an absence of CD4+ cells at the graft site without any effect on the cell composition in lymphatic tissues. CONCLUSION: Transgenic grafts that secrete anti-CD4 antibody can afford some protection against graft rejection, while only affecting the CD4 population at the graft site. PMID- 11014649 TI - Intragraft interleukin-4 mRNA expression after short-term CD154 blockade may trigger delayed development of transplant arteriosclerosis in the absence of CD8+ T cells. AB - BACKGROUND: It has recently been shown that, although anti-CD154 induces CD4+ T cell tolerance, it is unable to prevent allograft rejection mediated by CD8+ T cells. We have also shown that anti-CD154 monotherapy does not protect the graft from the development of transplant arteriosclerosis even in the absence of CD8+ T cells. This study was designed to investigate and characterize possible mechanisms responsible for the development of transplant arteriosclerosis after CD154 blockade in the absence of CD8+ T cells. METHODS: C57BL/6 (H2b) recipients received a fully MHC-mismatched BALB/c donor aorta (H2d). Animals were either treated with anti-CD154 monoclonal antibody (mAb) in the presence or absence of CD8 T cells. Histology, morphometric measurements, immunohistochemistry, and the production of alloantibodies (IgM, IgG1, IgG2a) were analyzed on days 14, 30, and 50 after transplantation. Cytokine production within the graft was investigated by competitive reverse transcription-polymerase chain reaction on day 14. RESULTS: Combined treatment with anti-CD154 and a depleting CD8 mAb resulted in a delay in the development of transplant arteriosclerosis (intimal proliferation: 33+/-10% vs. 67+/-11% untreated control, day 30) but ultimately did not prevent its progression (intimal proliferation: 55+/-10% vs. 78+/-9% untreated control, day 50). Although there was a significant decrease in the number of CD4+, CD11b+, and CD40+ graft-infiltrating cells and a reduction in the formation of donor specific IgG1 alloantibodies in recipients treated with anti-CD154 and anti-CD8 mAbs, mRNA for interleukin (IL)-4 was increased, suggesting a shift in the intragraft cytokine profile towards a Th2-like pattern. CONCLUSIONS: Our data provide evidence that short-term CD154 blockade is insufficient to prevent transplant arteriosclerosis, even in combination with CD8+ T-cell depletion. Moreover, the increased expression of the Th2 cytokine interleukin-4 within the graft may be responsible for the development of transplant arteriosclerosis in the long term. PMID- 11014650 TI - The pig analogue of CD59 protects transgenic mouse hearts from injury by human complement. AB - BACKGROUND: It has been proposed that hyperacute rejection (HAR) of pig-to primate vascularized xenografts is due in large part to ineffective regulation of recipient complement by pig complement regulatory proteins (CRPs), and indeed transgenic expression of human CRPs in pigs can prevent hyperacute rejection. However, at least one pig CRP (CD59) efficiently regulates human complement in vitro, suggesting that it is the level of expression of a particular CRP(s) rather than cross-species incompatibility that explains the HAR of porcine xenografts. We investigated the relative effectiveness of transgenically expressed pig and human CD59 in providing protection of mouse hearts from human complement in an ex vivo setting. METHODS: Transgenic mice expressing pig CD59 or human CD59 under the control of the human ICAM-2 promoter, which restricts expression in tissues to vascular endothelium, were used. Hearts from mice expressing similar levels of pig CD59 or human CD59 were perfused ex vivo with 10% human plasma and heart function was monitored for 60 min. Sections of perfused hearts were examined for deposition of the membrane attack complex (MAC). RESULTS: Control nontransgenic hearts (n=5) were rapidly affected by the addition of human plasma, with mean function falling to less than 10% of the initial level within 15 min. In contrast, hearts expressing either pig CD59 (n=6) or human CD59 (n=8) were protected from plasma-induced injury, maintaining 31 and 35% function, respectively, after 60 min of perfusion. MAC deposition was markedly reduced in both pig CD59 and human CD59 transgenic hearts compared to nontransgenic control hearts. CONCLUSIONS: When highly expressed on endothelium in transgenic mice, pig CD59 provided equivalent protection to human CD59 in a model of human complement-mediated xenograft rejection. Thus supranormal expression of endogenous porcine CRPs may be a feasible alternative to the expression of human CRPs in preventing HAR of pig-to-primate xenografts. PMID- 11014651 TI - Sirolimus (rapamycin) halts and reverses progression of allograft vascular disease in non-human primates. AB - BACKGROUND: Current immunosuppressive protocols fail to prevent chronic rejection often manifested as graft vascular disease (GVD) in solid organ transplant recipients. Several new immunosuppressants including sirolimus, a dual function growth factor antagonist, have been discovered, but studies of drug efficacy have been hampered by the lack of a model of GVD in primates, as a prelude to clinical trials. As described earlier, we have developed a novel non-human primate model of GVD where progression of GVD is quantified by intravascular ultrasound (IVUS). METHODS: Twelve cynomolgus monkeys underwent aortic transplantation from blood group compatible but mixed lymphocyte reaction-mismatched donors. To allow the development of GVD in the allograft, no treatment was administered for the first 6 weeks. Six monkeys were treated orally with sirolimus from day 45 after transplantation to day 105. RESULTS: Progression of GVD measured as change in intimal area from day 42 to 105 was halted in sirolimus-treated monkeys compared to untreated monkeys (P<0.001, general linear model). On day 105, the intimal area +/- SEM was 3.7+/-1.0 and 6.4+/-0.5 mm2, respectively (P<0.05, t test). The magnitude of allograft intimal area on day 105 correlated inversely with sirolimus trough levels (R2=0.67, P<0.05). Regression of the intimal area was seen in four of six sirolimus-treated monkeys, which was significantly different from the untreated monkeys (P<0.05). CONCLUSIONS: Our results in the first non human primate model of GVD showed that treatment with sirolimus not only halted the progression of preexisting GVD but also was associated with partial regression. Sirolimus trough blood levels were correlated with efficacy. Therefore, sirolimus has the potential to control clinical chronic allograft rejection. PMID- 11014652 TI - Single injection of insulin delays the recurrence of diabetes in syngeneic islet transplanted diabetic NOD mice. AB - BACKGROUND: Insulin has been implicated in the pathogenesis of type 1 diabetes and oral administration of insulin has been shown to delay the onset of diabetes in NOD mice. In this study we determined whether a single footpad injection of insulin will protect syngeneic islet grafts from autoimmune destruction when placed under the kidney capsule of diabetic NOD mice. METHODS: Five hundred islets were transplanted under the kidney capsule of diabetic female NOD mice in conjunction with a single footpad injection of either pork insulin in saline or mixed with incomplete Freund's adjuvant (IFA). Control groups received either IFA or saline alone. RESULTS: Seven of 11 animals (63.6%) given insulin in IFA exhibit long-term graft survival (>75 days; mean +/- SEM >85.4+/-16.1) whereas only 3 of 12 animals (25.0%) in the IFA group had graft survival longer than 75 days (mean +/- SEM >41.9+/-12.8 days). In contrast, none of the animals that received insulin in saline (17.3+/-2.5 days) and saline only (16.1+2.0 days) exhibit prolonged graft survival. CONCLUSION: These results suggest that a single footpad injection of insulin can protect the islet graft from immune attack in NOD mice. PMID- 11014653 TI - Tacrolimus-associated mutism after orthotopic liver transplantation. AB - BACKGROUND: Mutism/speech apraxia has been well documented as a toxic effect of cyclosporine after liver transplantation but has been reported only rarely with tacrolimus. Brain imaging with magnetic resonance or computed tomography has failed to demonstrate abnormalities in affected patients. METHODS: We present the first example of an acute onset of loss of speech associated with a sudden elevation of serum tacrolimus level after successful orthotopic liver transplantation. We also describe the positron emission tomography (PET) scan of this patient's brain. RESULTS: PET scan imaging of the brain was abnormal, demonstrating decreased metabolism in the posterior temporo-parieto-occipital regions. Statistical probability mapping revealed additional areas of hypometabolism in the cingulate gyrus. CONCLUSIONS: PET scan revealed abnormalities of the brain in a patient with tacrolimus-induced mutism. The cingulate gyrus may play a role in the mutism/speech apraxia syndrome seen with cyclosporine/tacrolimus neurotoxicity. PMID- 11014654 TI - Intrahepatic anastomosis formation between the hepatic veins in the graft liver of the living related liver transplantation: observation by Doppler ultrasonography. AB - In living related liver transplantation, the right lobe has come to be used as a graft to meet the metabolic demands of adult or adolescent recipients. In harvesting the right lobe as a graft, however, there is controversy as to whether or not the middle hepatic vein (MHV) should be included and reconstructed. Anatomical intrahepatic anastomosis between the right hepatic vein (RHV) and MHV is considered to exist, but the formation process of this functional anastomosis has not been demonstrated by Doppler ultrasonography (US). In our case, a right lobe including a right branch of the MHV was used as a graft. In implanting, the RHV was anastomosed to the inferior vena cava and the right branch of the MHV was ligated. Using Doppler US, we checked the blood flow in the hepatic vein after transplantation. Within 3 days of surgery, no flow was detected in the right branch of the MHV. A flow around the right branch of the MHV was observed at postoperative day 6. At postoperative day 9, a reverse flow was detected in which the right branch of the MHV drained into the RHV via the anastomosis between them. Based on our results, it appears that a functional intrahepatic anastomosis between hepatic veins formed gradually within 10 days of ligation of an afferent branch, during which time the graft function did not deteriorate. PMID- 11014655 TI - Managing incidents in the cervical screening programme. PMID- 11014657 TI - Fine needle aspiration cytology (FNAC) of adrenal masses. AB - Between 1992 and 1998 at the Pathology Section, Cytopathology Unit of the Department of Oncological and Surgical Sciences of the University of Padova, we performed one hundred and twelve FNACs of adrenal masses under radiological guidance and seven intraoperative scrape smears; histological follow-up was available in 55 cases. Immediate on-site assessment of smears revealed a satisfactory adequacy rate (92%). With a simple diagnostic tree, we have been able to classify all smears except one as benign or malignant correctly, (accuracy 97.6%), differentiation of primary tumours from metastatic depositions remaining the most difficult task. In our experience FNAC is a safe and accurate tool in the diagnostic characterization of adrenal masses. PMID- 11014656 TI - Analysis of proliferating cell fraction determined by monoclonal antibody to M1 subunit ribonucleotide reductase and Ki-67 in relation to p53 protein expression in fine-needle aspirates from non-Hodgkin's lymphomas. AB - The purpose of this study was to analyse the proliferative fraction with the monoclonal antibody M1-R-R to M1-subunit ribonucleotide reductase and with MIB-1 to Ki-67 antigen in relation to p53 protein expression in fine needle aspirates from B-cell non-Hodgkin's lymphomas. One hundred and thirty-seven cases, previously diagnosed and sub-typed according to the Kiel classification and characterized by immunophenotyping, were included in the study. The M-1 subunit ribonucleotide reductase (M1-R-R), Ki-67 and p53 antigens were detected using monoclonal antibodies on stored cytospin preparations. There was a good correlation (r = 0.72) between Ki-67 and M1-R-R positive cell fraction in both high and low grade lymphomas. High-grade lymphomas had a median percentage of M1 R-R/MIB-1 positive cells of 53.0/73.0 for lymphoblastic, 61.0/52.0 for immunoblastic and 33.5/41.0 for centroblastic lymphomas, respectively. In low grade lymphomas figures of median percentage of M1-R-R/MIB-1 were 9.0/15.0 for centroblastic/centrocytic, 11.0/9.5 for chronic lymphocytic leukaemia, 16.0/27.0 for centrocytic and 12.0/9.0 for immunocytomas, respectively. The median percentages of M1-R-R/MIB-1 for high and low grade lymphomas were 37.0/50.5 and 11.0/12.0, respectively. In the p53 positive cases the proliferation rate as measured by staining for M1-R-R and MIB-1 was higher than in p53 negative cases, but the difference was not statistically significant. The results show that cytospin material obtained by fine needle aspiration and stored at -70 degrees C for years can be used reliably for both peroxidase-avidin-biotin and three-step alkaline phosphatase immunocytochemical staining. In addition, proliferation fraction determined by M1-R-R monoclonal antibody staining correlates well with that measured by an established marker for cell proliferation, the Ki-67 antibody. However, the proliferation fraction as measured by the two antibodies differs in the various subtypes of non-Hodgkin's lymphoma which indicates that they may contribute different prognostic information. PMID- 11014658 TI - The usefulness of a panel of immunostains in the diagnosis and differentiation of metastatic malignancies in pericardial effusions. AB - Pericardial effusions are not uncommon in patients with an advanced malignancy Rarely malignancies may present initially with a pericardial effusion. Cytological examination of pericardial fluid may be valuable in differentiation of these cases. However, a metastatic tumour in serous effusion may not always show the functional differentiation of the primary tumour. In such a situation, although a wide range of special studies have been suggested for the diagnosis of malignancy we have found the use of a panel of a few common immunostains to be useful in confirming or suggesting the site of a primary tumour. The material for this study consisted of 76 pericardial fluids obtained between January 1991 and October 1998 from 46 males (mean age 59 years) and 30 females (mean age 52 years). Metastatic malignancy was diagnosed in 22 of the 76 patients and in 7/22 cases pericardial effusions were the initial presentation. The subsequent follow up in the seven cases revealed adenocarcinoma of lung (n = 2), small cell anaplastic carcinoma of lung (n = 1), squamous cell carcinoma lung (n = 1), melanoma leg (n = 1), non-Hodgkin's lymphoma retroperitoneal lymph nodes (n = 1) and carcinoma of the breast (n = 1). Of the remaining 15 cases with a known history of malignancy, eight had cancers (three adeno; two small cell; one poorly differentiated, and two squamous cell types) of the lung; breast (n = 3); colon (n = 1); melanoma (n = 2) and non Hodgkin's lymphoma (n = 1). Immunostains which were useful in the diagnosis were EMA, CEA, cytokeratin, B72.3, HMB45, vimentin, S100, LCA, L26 and kappa and lambda light chains. PMID- 11014659 TI - Exact Touch: a new device for cervical cytology. Comparison with Ayre spatula plus Cytobrush. AB - Exact Touch is a new plastic device for the collection of cells from the ectocervix and endocervix. A total of 189 consecutive women were evaluated, 94 with the Ayre/cytobrush and 95 with Exact Touch. Sampling was performed by only one clinician, and the slides were analysed by only one cytotechnologist, who had no information about the sampling method. Our results showed that more endocervical and metaplastic cells were collected by Exact Touch than by Ayre/cytobrush. PMID- 11014661 TI - Continuing education for cytotechnologists: the Australian experience. AB - The development of a continuing education scheme for cytotechonologists in Australia is described. The process involved the establishment of a working party with Terms of Reference to review current practice in Australia and existing schemes in other parts of the world. The scheme developed takes the form of a continuing education diary that provides guidelines on the various forms of continuing education activity and corresponding credit points. The diary also provides for a record of activity to be kept. The scheme requires bi-annual submission of personal activity which is logged into a national database. A peer profile is provided and successful achievement is marked by the issue of a certificate of participation. The programme has achieved a 57% compliance in its first year of operation. PMID- 11014660 TI - The interaction of trichomonas vaginalis with epithelial cells, polymorphonuclear leucocytes and erythrocytes on vaginal smears: light microscopic observation. AB - In this study, vaginal smears taken from 400 patients were examined cytologically using the Papanicolaou technique. Twenty of the 400 patients were detected as harbouring Trichomonas vaginalis. The interactions of T. vaginalis with epithelial cells, polymorphonuclear leucocytes (PMNLs) and erythrocytes were determined at light microscopic level. It was observed that T. vaginalis were juxtaposed to the epithelial cells and changed shape according to the contours of the epithelial cell revealing the cytopathic effect of trichomonads on epithelial cells. Trichomonads attached to PMNLs produced pseudopodia to phagocytose the cells. Occasionally an amoeboid shaped T. vaginalis organism was seen trailed by a row of PMNLs. This light microscopic study supports the production by trichomonads of a chemotactic factor for PMNLs. Phagocytosed erythrocytes were also seen in the cytoplasm of T. vaginalis, suggesting the need for the patient to be tested for anaemia. PMID- 11014662 TI - Educational case report. Cohesive cells in a breast fine needle aspirate. PMID- 11014663 TI - Endosalpingiosis (mullerianosis) of the bladder: a potential source of error in urinary cytology. PMID- 11014664 TI - Reprocessing inadequate breast FNAC 'Cytospin' samples to improve diagnostic rates--an audit. PMID- 11014665 TI - George Papanicolaou (1883-1962). PMID- 11014666 TI - Acinic cell carcinoma arising in pleomorphic adenoma of parotid gland. PMID- 11014667 TI - A model for experience-dependent changes in the responses of inferotemporal neurons. AB - Neurons in inferior temporal (IT) cortex exhibit selectivity for complex visual stimuli and can maintain activity during the delay following the presentation of a stimulus in delayed match to sample tasks. Experimental work in awake monkeys has shown that the responses of IT neurons decline during presentation of stimuli which have been seen recently (within the past few seconds). In addition, experiments have found that the responses of IT neurons to visual stimuli also decline as the stimuli become familiar, independent of recency. Here a biologically based neural network simulation is used to model these effects primarily through two processes. The recency effects are caused by adaptation due to a calcium-dependent potassium current, and the familiarity effects are caused by competitive self-organization of modifiable feedforward synapses terminating on IT cortex neurons. PMID- 11014668 TI - Independent component analysis applied to feature extraction from colour and stereo images. AB - Previous work has shown that independent component analysis (ICA) applied to feature extraction from natural image data yields features resembling Gabor functions and simple-cell receptive fields. This article considers the effects of including chromatic and stereo information. The inclusion of colour leads to features divided into separate red/green, blue/yellow, and bright/dark channels. Stereo image data, on the other hand, leads to binocular receptive fields which are tuned to various disparities. The similarities between these results and the observed properties of simple cells in the primary visual cortex are further evidence for the hypothesis that visual cortical neurons perform some type of redundancy reduction, which was one of the original motivations for ICA in the first place. In addition, ICA provides a principled method for feature extraction from colour and stereo images; such features could be used in image processing operations such as denoising and compression, as well as in pattern recognition. PMID- 11014669 TI - Diffusion and innovation rates for multidimensional neuronal data with large spatial covariances. AB - The diffusion model has been introduced as a statistical model for processing multidimensional neuronal data. This paper extends estimation procedures for the parameters of this model when spatial covariances are large. The new method is based upon linear regression techniques. It is applied to an optical recording of the auditory cortex of a guinea pig stimulated with pure tone bursts (frequency 14 kHz). PMID- 11014670 TI - Effects of delay on the type and velocity of travelling pulses in neuronal networks with spatially decaying connectivity. AB - We study a one-dimensional model of integrate-and-fire neurons that are allowed to fire only one spike, and are coupled by excitatory synapses with delay. At small delay values, this model describes a disinhibited cortical slice. At large delay values, the model is a reduction of a model of thalamic networks composed of excitatory and inhibitory neurons, in which the excitatory neurons show the post-inhibitory rebound mechanism. The velocity and stability of propagating continuous pulses are calculated analytically. Two pulses with different velocities exist if the synaptic coupling is larger than a minimal value; the pulse with the lower velocity is always unstable. Above a certain critical value of the constant delay, continuous pulses lose stability via a Hopf bifurcation, and lurching pulses emerge. The parameter regime for which lurching occurs is strongly affected by the synaptic footprint (connectivity) shape. A bistable regime, in which both continuous and lurching pulses can propagate. may occur with square or Gaussian footprint shapes but not with an exponential footprint shape. A perturbation calculation is used in order to calculate the spatial lurching period and the velocity of lurching pulses at large delay values. For strong synaptic coupling, the velocity of the lurching pulse is governed by the tail of the synaptic footprint shape. Moreover, the velocities of continuous and lurching pulses have the same functional dependencies on the strength of the synaptic coupling strength gsyn: they increase logarithmically with gsyn for an exponential footprint shape, they scale like (In gsyn)1/2 for a Gaussian footprint shape, and they are bounded for a square footprint shape or any shape with a finite support. We find analytically how the axonal propagation velocity reduces the velocity of continuous pulses; it does not affect the critical delay. We conclude that the differences in velocity and shape between the front of thalamic spindle waves in vitro and cortical paroxysmal discharges stem from their different effective delays. PMID- 11014671 TI - Optic nerve evaluation among optometrists. AB - PURPOSE: To determine agreement among optometrists regarding assessment of optic nerve C/D ratios and perceived glaucomatous damage and to separately analyze these results on the basis of residency training, practice setting, and glaucoma patient experience. METHODS: Fifty-six optometrists from various modes of professional practice evaluated 33 stereoscopic optic nerve photographs. Observers were asked to estimate the vertical cup-to-disk (C/D) ratio and determine the glaucomatous status of the optic nerve. The mean vertical C/D ratio, percentage perceived as glaucomatous, and levels of interobserver and intraobserver agreement (kappa) are reported. RESULTS: Estimated C/D ratios differ significantly on the basis of training (p = 0.02) practice setting (p = 0.001), glaucoma patient experience (p = 0.001). Glaucomatous damage interpretation was significantly different (p = 0.006) based upon an optometrist's practice setting. Interobserver agreement regarding C/D ratios is significantly higher among optometrists who have completed a residency (kappaw = 0.59) and practice in clinical settings (optometry school/medical center/hospital) (kappaw = 0.59) compared with non-residency-trained optometrists (kappaw = 0.52) and those practicing in commercial settings (kappaw = 0.54). Interobserver agreement of glaucomatous damage is significantly higher among optometrists who have completed a residency (kappa = 0.50) compared with non residency-trained optometrists (kappa = 0.42). CONCLUSIONS: Intraobserver agreement is higher than interobserver agreement among optometrists when C/D ratios are estimated and the glaucomatous status of the optic nerve is assessed. Optic nerve evaluation among optometrists is significantly influenced by residency training, practice setting, and glaucoma patient encounters. PMID- 11014672 TI - Field expansion for homonymous hemianopia by optically induced peripheral exotropia. AB - PURPOSE: To describe a novel method for prism correction of hemianopia that provides field-of-view expansion in a convenient and functional format and to evaluate initial clinical application. METHOD: To expand the upper quadrant of the field, a high power prism segment (30-40delta) is placed base-out across the upper part of the spectacle lens, on the side of the loss, at about the level of the limbus. A similar prism segment at the lower part of the lens is used to treat the lower field. The peripheral location of the prisms causes peripheral exotropia. As a result a scene segment as high as the vertical span of the prism is shifted laterally by 15 to 20 degrees relative to the view of the other eye. At the edge of the hemianopic field loss, objects that would fall in the scotoma of one eye are seen through the prism in the other eye, providing a simultaneous awareness of details within the otherwise absent field-of-view. An approach for fitting the system to patients with abnormal binocular vision (strabismus and amblyopia, with or without diplopia) is discussed as well. The effect of the prisms was evaluated in a noncomparative case series (12 patients). RESULTS: The field expansion is provided at any position of lateral gaze, including gaze away from the side of the scotoma. The effect of this technique on field expansion was demonstrated using standard binocular perimetry. Most patients reported substantial improvement in function and in obstacle avoidance. CONCLUSION: A novel method for the optical treatment of hemianopia was developed and tested. It was found to be effective in expanding the field and helping patients' mobility. PMID- 11014673 TI - Temporal variations in myopia progression in Singaporean children within an academic year. AB - PURPOSE: Excessive nearwork is believed to be associated with myopia development and progression. To investigate this further, we studied refractive error changes and their correlation with nearwork in a cohort of grade school children in Singapore. METHODS: Cycloplegic autorefraction was performed 5 times over 10 months on 168 children aged 7, 9, and 12 years who were further divided into myopic and nonmyopic subgroups based in their initial refractive errors. Information about nearwork was obtained through diaries filled out over 24 h at the commencement of the study. RESULTS: Myopia progression was high (overall mean: -0.87 D per year) and largely linear throughout the year, but significantly higher rates were seen after the final school examinations in 7-year-old myopes and nonmyopes. Overall, myopic groups exhibited higher progression rates than nonmyopic groups, although 33.6% of subjects from the latter groups had become myopic by the end of the study. Nearwork scores derived from the diaries were generally not well correlated with overall myopia progression. CONCLUSIONS: The tendency for myopia progression rates to increase after the final school examinations in 7-year-olds is interpreted as a delayed effect of the intense nearwork associated with preparing for them. The timing of nearwork-diary data collection at the beginning of the study could be responsible for the poor correlation between these data and overall myopia progression rates. PMID- 11014674 TI - Inter-rater and intra-rater reliability in the interpretation of MTI Photoscreener photographs of Native American preschool children. AB - PURPOSE: To evaluate inter- and intra-rater reliability for the interpretation of MTI Photoscreener photographs taken in a population of Native American preschool children with a high prevalence of astigmatism. METHODS: Photographs of 369 children were rated by 11 nonexpert and 3 expert raters. Photographs for each child were scored as pass, refer, or retake. Nonexpert raters scored photos on two separate occasions, permitting analysis of intra-rater reliability. RESULTS: Analyses of pass/refer responses only: inter-rater reliability was moderate to substantial among nonexpert raters and substantial among expert raters. Intra rater reliability among nonexperts was substantial. Analyses of all responses (pass, refer, and retake): inter-rater reliability for pass and refer scores was moderate among nonexperts and substantial among experts; for retake scores inter rater reliability was slight for nonexperts and moderate for experts. Intra-rater reliability among nonexperts was substantial for pass and refer scores and moderate for retake scores. CONCLUSIONS: In this population with a high prevalence of astigmatism, whether MTI photoscreening results are interpretable is much more variable among and within raters than whether an interpretable photograph should be scored as pass or refer. The level of agreement among raters in the current study was influenced by the experience of the raters. In addition, nonexpert raters were more likely to deem a photograph uninterpretable than expert raters. PMID- 11014675 TI - The accuracy and precision performance of four videokeratoscopes in measuring test surfaces. AB - In this study we evaluated the accuracy and precision of three placido-disk videokeratoscopes (the Keratron, Medmont and TMS) and one videokeratoscope that uses the raster-stereogrammetry technique (PAR-CTS) in elevation topography with six test surfaces. The test surfaces were a sphere, an asphere, a multicurve, and three bicurve surfaces. Each instrument performed well on certain test surfaces, but none of the instruments excelled on all of the surfaces. The results showed high accuracy of the Keratron and Medmont instruments in measuring the sphere, asphere, and multicurve surfaces, but not the bicurve surfaces. The precision of the Keratron and Medmont instruments were high. The TMS and PAR-CTS instruments showed poorer accuracy than the Keratron and Medmont instruments for the multicurve test surface but showed better performance for the bicurve surfaces. The PAR-CTS had the poorest performance in precision of the four instruments. The use of the Noryl spherical test surface instead of polymethyl methacrylate (PMMA) resulted in small differences in the accuracy performance of the placido-disk videokeratoscopes only. PMID- 11014676 TI - Performance on the three-point Vernier alignment or acuity test as a function of age: measurement extended to ages 5 to 9 years. AB - The three-point Vernier alignment (or acuity) test was conducted on children from 5 to 9 years old. There is a significant difference between the 5-9 year old subjects and those in age groups 10 to 19 and 20 to 29 years in Vernier performance. These data were also compared with previously published data from older subjects (up to age 70). We conclude that Vernier function has not fully matured within the age range of 5 to 9 years; this finding is consistent with previous results reported in the literature. PMID- 11014677 TI - Heart and lung support interaction--modeling and simulation. AB - Mechanical support of the lungs used to preserve life or during any kind of surgery may have an adverse effect on the cardiovascular system. Usually, positive pressure in alveoli diminishes lung perfusion, venous return and cardiac output. Positive pressure during the respiratory cycle is transfered into the thoracic space. The aim of this study was to assess how synchronization of the respirator with spontaneous breathing influences the distribution of pressure and ventilation in nonhomogeneous lungs and how it should influence hemodynamics. For this purpose a multicompartmental model of respiratory system mechanics was used in the electrical analog of a respirator-lung circuit, which enabled us to simultaneously simulate ventilatory support and spontaneous breathing. Mechanical properties of the respiratory system were modeled by lumped parameters: resistances and capacitances of constant values, independent of lung volume or inspiratory flow changes. A multicompartmental model of the respiratory system enabled us to simulate lung pathology characterized by non-homogeneity of the mechanical properties of the different parts of the lungs. The results of simulations presented in the paper enable us to conclude that lung volume increase, independent of the respirator-patient breathing synchronization, may be modeled as the increase in pulmonary vascular resistance and alveolar pressure increase, dependent on respirator-patient breathing synchronization, may be averaged by esophageous balloon measurements which show intrathoracic pressure changes. PMID- 11014678 TI - Surface pitting of heart valve disks tested in an accelerated fatigue tester. AB - There are various reports on the fracture of mechanical heart valves implanted in humans or animals and it has been pointed out that fractures are induced by erosion of the disk surface due to cavitation bubbles. Cavitation erosion on mechanical heart valves was studied using our originally designed accelerated fatigue tester. Several valve housings with different compliance values were used. The number and position of pits on the valve disk were measured using an optical microscope. Disk-closing velocity was measured and cavitation bubbles were monitored by a high-speed video camera. It was found that disk-closing velocity increased and cavitation erosion was enhanced with an increase in compliance of the valve holder. Therefore, careful attention should be paid to the compliance of an accelerated fatigue tester. PMID- 11014679 TI - Artificial ventilation of the lungs for emergencies. AB - The necessity for extraordinary ventilatory support may appear in different places all over the world in cases of a massive disaster (industrial or natural), connected with gas poisoning on a huge scale. Hospitals equipped with limited number of respirators, adequate for peacetime activity, are not able to meet suddenly multiplied requirements for ventilatory support. This paper describes a preliminary study to develop a convenient, reliable method of performing artificial ventilation of at least two patient by means of only one ventilator. We developed a unique, new control system (patent pending) which, when placed between a respirator and endotracheal tubes of the patients, divides the total tidal volume between the patients' lungs and controls pressures at their airways. A special arrangement of valves in the control system enables us to separate inspiratory and expiratory paths for each patient and to avoid cross-infection. The model study performed, according to ISO standards, on mechanical test lungs has shown that the proposed control system enables us to adjust ventilatory parameters at desired values, when lung compliance or respiratory airway resistance differ. The proposed one-source artificial ventilation is a simple solution to provide ventilatory support when the number of patients is greater than the number of respirators that are available. PMID- 11014680 TI - Oximetric and capnometric studies--aspects of natural and artificial ventilation. AB - Methods and instrumentation of own design and their application to the measurement of oxygen and CO2 content in blood and respiration gases are discussed. Spectrophotometry, particularly double wavelength colorimetry using red light and IR light, applied to the measurement of oxygen blood saturation for in vitro examination and through an intravascular catheter for invasive in vivo study and by means of pulse oximetry for non-invasive in vivo examination are presented. The electrochemical methods applied to in vitro and in vivo measurement of oxygen in blood (amperometric method) and in respiration gases (amperometric and fuel cell methods) are discussed. The electrochemical Stow Severinghaus method of CO2 tension measurement in blood and CO2 partial pressure in respiratory gases as well as the selective IR light (4.26 microm) absorption method and instruments of own design for CO2 measurements in respiratory gases are also discussed. The methods and instrumentation described are very important for monitoring respiratory and circulatory function, particularly during artificial ventilation and circulatory support. PMID- 11014681 TI - Three methods for investigating, assessing and supporting the normal and pathological gait and manipulation activity. AB - Three methods to describe biped locomotion as well as manipulation activity are presented. Methods using so-called couple oscillators as rhythm generators are described. Based on a mathematical model, results of computer simulation of biped locomotion are presented. An indices method of assessing human gait for both normal and pathological cases was proposed and tested. The results showed that it is relatively easy to distinguish different gaits by using this method. This method was tested in clinical conditions. The third method deals with the problem of supporting the lost function of prehension movements. Implanted stimulators on the nerves combined with external orthoses were used. The clinical results show that these methods have practical advantages. PMID- 11014682 TI - Computer-aided generation of stimulation data and model identification for functional electrical stimulation (FES) control of lower extremities. AB - Standard stimulation data for unassisted standing up of paraplegic patients was generated by dynamic optimization linked with model simulation, to overcome the difficulties in the present electromyogram (EMG)-based method. The generated stimulation data were roughly in agreement with the normal subjects' EMG. From these, it is suggested that the 'model-based' method is useful as an alternative of the 'EMG-based method'. The same technique can be applied to generation of patient-specific stimulation data once the musculoskeletal system of a patient is properly identified. The musculoskeletal system must be identified from data taken from simple and noninvasive experiments for the identification method to be practically acceptable. We developed a musculoskeletal model and systematic identification protocols for this purpose. They were validated for the vastus lateralis muscle at the knee joint. The identification was successful and the predicted joint angle trajectories closely matched the experimental data. This implies that the model-based generation of patient-specific stimulation data is possible. PMID- 11014683 TI - Manifestation of internal organs malfunction by laser Doppler study on microcirculation. AB - Monitoring of microvascular blood perfusion provides very specific information on the proper function or malfunction of some internal organs, e.g. the pancreas and kidney. The laser Doppler method was used to measure microperfusion in the skin of the lower limb of diabetic patients and patients undergoing hemodialysis. This method offers non-invasive, real-time monitoring and is already accepted in many clinical experiments. The method and the laser Doppler instruments used are described. Special attention is paid to the investigation of microvascular abnormality in diabetes by using a multichannel laser Doppler system during postocclusive reactive hyperemia. The study group consisted of 41 diabetes and 24 healthy subjects with no history of family diabetes. The most valuable data were obtained from the probe located on the most distal part of the foot. Some hyperemic parameters (maximum of hyperemic response, time to peak flow) were significantly different for the diabetic group as comparing to the norm. In the microcirculation study during hemodialysis, three patients with kidney dysfunction were investigated. Increase in red blood cell velocity was observed, probably caused by better distribution of blood to the peripheral circulation. PMID- 11014684 TI - Unconstrained physiological monotoring in daily living for health care. AB - This paper deals with the recent development of two types of non-invasive physiological monitoring systems for possible application in an unconstrained manner to normal subjects for health care as well as to patients and/or outpatients with disorders or with life support systems (artificial organs and organ transplantations). One is an ambulatory monitoring system which allows automatic acquisition of blood pressure, cardiac output and other cardiovascular hemodynamic parameters on a beat-by-beat basis using the volume-compensation and transthoracic electrical admittance method. The other is a home monitoring system installed in a lavatory which can measure body and excreta weight together with the ballistocardiogram as an index of cardiac ejecting function in an unaware fashion without attachment of any sensors to the subject's body and without special operations for measurement during toilet use. Outlines of these two systems and monitoring results of laboratory and field testings are presented, and these suggest that the ambulatory and non-conscious physiological monitoring techniques described herein appear promising as a valuable and helpful means for use in research as well as in the practical field of health monitoring at home during daily living. PMID- 11014685 TI - Acquisition of long-term cardiac signals for chronodiagnostic utility. AB - Periodical and long-term examination of the cardiac function is important for detecting early-stage disorders, and contributing to chronodiagnosis and chronotherapeutics. Frequent or daily examination utilizing the present test procedures, however, places the patient under stress even when it is administered at home and not during a hospital visit. In order to cope with this issue, monitoring systems whereby a patient is not aware of being examined were developed taking the electrocardiogram (ECG) as an example. The ECG was monitored while the patient was taking a bath or sleeping in bed. The system did not utilize any body surface electrodes but introduced in-water electrodes in the bath and textile electrodes on the bed. These systems made long-term observation feasible without imposing any stress on the patient. PMID- 11014686 TI - Dialyzer reuse: what we know and what we don't know. AB - Despite extensive clinical experience, the effects of different reuse procedures have not been fully evaluated. The available data suggest that the effect of reuse on dialyzer performance depends upon the type of chemicals employed, the membrane type, and the size of the solute whose removal is being assessed. The effect of reuse on urea clearance is essentially defined by the residual cell volume with a total cell volume of > 80% associated with a dialyzer clearance that is within 10% of its original value. The effect of reuse on large solute clearance can be dramatic, with the procedure resulting in substantial changes in the beta2-microglobulin clearance of different dialyzers. Of note is the limited data available regarding the effect of reuse procedures on dialyzers processed more than 20 times. PMID- 11014687 TI - Are parathyroidectomies still appropriate in chronic dialysis patients? AB - The uremic milieu generates chronic stimulatory input to the parathyroid glands, which is mediated principally by low calcium, high phosphate and low calcitriol, and results in increased parathyroid hormone (PTH) synthesis and release and an increase in parathyroid mitotic activity with the development of monoclonal areas of nodular hyperplasia. Such glands do not fully express the machinery required to mediate the suppressive inputs to the parathyroids; the extracellular calcium receptor (CaR) and the vitamin D receptor (VDR) are both downregulated. In most of these patients ablation, by parathyroidectomy or ethanol injection, provides the only means of correcting the hyperparathyroidism; apoptosis in parathyroid cells is negligible and clinically irrelevant. In practice, surgery is often delayed by a doomed and ultimately futile attempt to effect control by medical means. Better predictors of the likely success or failure of optimal non surgical management are needed. Gland size exceeding 1 cm3 and elevated PTH despite hypercalcemia (implying loss of suppressibility by calcium), in the presence of good phosphate control and adequate calcitriol provision point strongly to eventual failure of medical treatment and the need for parathyroid ablation. Parathyroidectomy, usually subtotal, remains the standard management, with ultrasound guided injection of ethanol or calcitriol showing promise in some centers. The above scenario is unlikely to be changed greatly by the new emerging vitamin D metabolites, but calcimimetic agents may well increase the scope of non surgical management. PMID- 11014688 TI - Non-dialytic treatment of acute hyperkalemia in the dialysis patient. AB - Acute hyperkalemia occurs commonly amongst patients with chronic renal failure and is especially common in noncompliant individuals. These patients often present to the emergency room with weakness. This editorial addresses the issue of non-dialytic treatment of hyperkalemia. Is emergency dialysis indicated in every case? In my opinion, acute dialysis can often be delayed until the hospital's dialysis unit opens for "regular working hours." PMID- 11014689 TI - Does reuse have clinically important effects on dialyzer function? PMID- 11014690 TI - Strategy for maximizing the use of arteriovenous fistulae. AB - Increasing the use of arteriovenous fistulae in dialysis patients requires a specific strategy. In order to properly select patients for an arteriovenous fistula (AVF), it is essential that the nephrologist become knowledgeable about the subject and that an organized approach be followed. Both the arterial and venous systems must be evaluated. Evaluation of medical history, general physical examination, specific physical examination related to the vasculature of the extremity, vein mapping and duplex ultrasound studies are all important. It is very important to assess the size of the vessels involved. Although a newly created AVF should be allowed to fully mature prior to use, failure to develop should be evaluated early. Many cases of early failure can be successfully salvaged. Even if the patient has an arteriovenous graft, they should be evaluated at the time of every graft failure for the possibilities of creating a secondary AVF. PMID- 11014691 TI - Peritoneal sclerosis: one or two nosological entities? AB - The frequency, pathology, animal models, pathogenesis, clinical manifestations, diagnostic criteria, therapy and prevention of peritoneal sclerosis are reviewed. Many of these aspects have a bimodal configuration which suggests that peritoneal sclerosis, usually considered a single pathology in peritoneal dialysis, is actually two distinct nosological entities: simple sclerosis and sclerosing peritonitis. The former is very frequent, with minor anatomical alterations and low clinical impact; it is reproducible in animals by means of peritoneal dialysis, and is clearly due to the poor biocompatibility of peritoneal dialysis solutions. The latter is rare, with radical anatomical alterations and high mortality requiring valid methods of diagnosis, therapy and prevention; it can only be reproduced in animal models by means other than peritoneal dialysis and seems to be due to factors both related and unrelated to peritoneal dialysis. PMID- 11014692 TI - Hemodialyzer membranes and configurations: a historical perspective. AB - The principle of hemodialysis (HD) was first described over a century ago while the first human HD treatment was performed in 1923 with collodion tubes. Since that time, a variety of different hemodialyzer configurations and membranes have been used. The purpose of this article is to provide a historical review of these various configurations and membranes. The rotating drum, coil, parallel flow, and hollow fiber artificial kidneys are discussed. Emphasis is placed on the factors that have influenced the shaping of the contemporary HD market. PMID- 11014693 TI - Upper limb ischemia after vascular access surgery: differential diagnosis and management. AB - Hand ischemia following placement of upper limb arteriovenous accesses for dialysis may result in debilitating complications and contribute to morbidity and mortality on dialysis. Two distinct clinical variants of hand ischemia are recognized: vascular steal syndrome, in which a spectrum of severity of ischemic changes affect all tissues of the hand; and ischemic monomelic neuropathy, where ischemia is confined to the nerves of the hand. Early diagnosis and treatment of these complications (often including closure of the access) is imperative to prevent hand paralysis, and digital or hand amputation. PMID- 11014694 TI - Clinical and economic outcomes of the use of expanded criteria donors in renal transplantation. PMID- 11014695 TI - Comorbidity assessment in hemodialysis and peritoneal dialysis using the index of coexistent disease. AB - The purpose of this paper is to describe the ICED, summarize outcomes of prior studies in which it was used, and describe the adaptations that have lead to the present instrument. We will then demonstrate its use in quantifying the burden of comorbid conditions in a sample of hemodialysis and peritoneal dialysis patients from our center, and show the relationship between ICED levels and outcomes in peritoneal dialysis patients. PMID- 11014696 TI - Chloramine-induced methemoglobinemia in a hemodialysis patient. PMID- 11014697 TI - Acquired cystic kidney disease. PMID- 11014698 TI - Intravenous vitamin C for erythropoietin resistance. PMID- 11014699 TI - Safety of high venous and arterial line pressures during hemodialysis. PMID- 11014700 TI - Causes of lability in PTH values. PMID- 11014701 TI - Characteristics of peritonitis in HIV-positive PD patients. PMID- 11014702 TI - Rationing dialysis: the early years. PMID- 11014703 TI - Measured and predicted normalized peritoneal clearances. PMID- 11014704 TI - Primary and secondary control strategies for managing health and financial stress across adulthood. AB - The study examined the relation among three types of control strategies (persistence, positive reappraisals, lowering aspirations) and subjective well being across adulthood (N = 3,490). Specifically, the authors investigated whether age-adapted endorsement of control strategies is conducive to subjective well-being if individuals experience health or financial stress. The results reveal an overall enhanced reliance on control strategies in older as compared with younger adults. In addition, persistence showed a stronger positive relation to subjective well-being in young adulthood as compared with old age. In midlife and old age, positive reappraisals had a stronger positive relation to subjective well-being than persistence. Lowering aspirations was negatively related to subjective well-being, independent of age. Age differences in the relation of control strategies to subjective well-being were particularly salient in individuals who faced either health or financial stress. PMID- 11014705 TI - Personal theories, intellectual ability, and epistemological beliefs: adult age differences in everyday reasoning biases. AB - Age-related differences in everyday reasoning biases were explored. In each of 2 social domains, examination of theoretical beliefs and biases along 2 dimensions of scientific reasoning, involving the law of large numbers and the evaluation of experimental evidence, revealed that, across age groups, scientific reasoning was used to reject evidence that contradicted prior beliefs; relatively cursory reasoning was used to accept belief-consistent evidence. Biased reasoning was more common among middle-aged and older adults than among young adults. Dispositions to engage in analytic processing were negatively related to biases, but intellectual abilities and bias were not related. The findings support a 2 process view of adult cognitive development and suggest that the tendency to rely on heuristic information processing increases with age. PMID- 11014706 TI - Memorizing while walking: increase in dual-task costs from young adulthood to old age. AB - The dual task of memorizing word lists while walking was predicted to become more difficult with age because balance and gait are in greater need of "attentional resources." Forty-seven young (ages 20-30 years), 45 middle-aged (40-50), and 48 old (60-70) adults were trained to criterion in a mnemonic technique and instructed to walk quickly and accurately on 2 narrow tracks of different path complexity. Then. participants encoded the word lists while sitting, standing, or walking on either track; likewise, speed and accuracy of walking performance were assessed with and without concurrent memory encoding. Dual-task costs increased with age in both domains; relative to young adults, the effect size of the overall increase was 0.98 standard deviation units for middle-aged and 1.47 standard deviation units for old adults. It is argued that sensory and motor aspects of behavior are increasingly in need of cognitive control with advancing age. PMID- 11014707 TI - Modeling caregiver adaptation over time: the longitudinal impact of behavior problems. AB - Although cross-sectional research has established the link between care demands and various indicators of caregiver adaptation, few studies have examined the impact of care recipients' problematic behavior over time. The present analysis determines the importance of behavior problems when predicting rates of change in subjective stressors (role overload and role captivity) and depression. Using 4 wave longitudinal data (N = 137) on dementia caregivers, the authors fit individual growth curve models for care demands (i.e., behavior problems, activities of daily living dependencies, and cognitive impairment), subjective stressors, and depression. Subsequent structural equation models found that increases in behavior problems were most likely to predict increases in role overload. The findings emphasize the deleterious long-term impact of behavior problems on individuals' emotional adaptation to caregiving. PMID- 11014708 TI - Age differences in feeling-of-knowing and confidence judgements as a function of knowledge domain. AB - For two semantic knowledge domains, general and computer-related, feeling-of knowing (FOK) and confidence level (CL) ratings and their relative accuracy were assessed in young, middle-aged, and older adults, after test difficulty was equated across age groups. Global memory self-efficacy beliefs were also assessed for each domain. As expected, greater age was associated with poorer memory self efficacy beliefs only in the computer domain. The oldest two groups were found to be more underconfident than young adults when rating their FOK but not their CL, for computer items but not for general items. Statistical control of age differences in memory self-efficacy beliefs in the relevant domain greatly reduced this age effect on computer-related FOK ratings. This finding suggests that absolute FOK judgments are more closely related to memory self-efficacy beliefs than are CL judgments. Gamma correlations between judgments and recognition performance revealed that all age groups were equally accurate in FOK and in CL judgments, in both domains. PMID- 11014709 TI - Updating knowledge about encoding strategies: a componential analysis of learning about strategy effectiveness from task experience. AB - Researchers have argued for age deficits in learning about the effects of encoding strategies from task experience, partly on the basis of absolute accuracy of metacognitive judgments. However, these findings could be attributed to factors other than age differences in learning. Forty older and 40 younger adults participated in 2 study-test trials in which they studied paired associates with imagery or repetition, predicted recall for the items, attempted recall, and postdicted recall. Recall was greater after imagery than repetition, yet this effect was not fully reflected by predictions made on Trial 1. Although both older and younger adults accurately postdicted recall from Trial 1, absolute accuracy of the predictions made on Trial 2 showed little improvement. By contrast, both age groups demonstrated increases in between-person correlations of predictions with recall, which is inconsistent with age deficits in knowledge updating. Thus, both older and younger adults had updated knowledge about the relative effects of the strategies, but such updating was not evident in the absolute accuracy of the predictions. PMID- 11014710 TI - Unwanted thought: age differences in the correction of social judgements. AB - This study use a false information paradigm to study age differences in the correction of social judgments. Younger and older adults read 2 criminal reports, with true information printed in black and false information in red. Following the reports, all participants were asked to recommend prison terms among other ratings. Age differences in baseline measures were also assessed by corresponding control groups who read only true information. Compared with younger adults under full attention, older adults under full attention and younger adults under divided attention were reliably influenced by the nature of the false statements (either extenuating or exacerbating the severity of the crimes). When contrasted with their relevant control groups, older adults under full attention and younger adults under divided attention failed to correct their social judgments. This study lends support to a processing resource explanation for age differences in the correction process for social judgments. PMID- 11014711 TI - Measures of fluency as predictors of incidental memory among older adults. AB - This study investigated fluency performance as a mediator of age-related declines in incidental memory performance as both are thought to rely on strategic retrieval processes. A large sample of community dwelling older adults completed a battery of tests assessing fluency, verbal knowledge, speed of information processing, and incidental recall. Fluency measures included initial and excluded letter fluency and the Uses for Objects Test, and they were assumed to reflect increasing reliance on strategic retrieval search. Speed emerged as the best mediator of age-related variance in incidental recall, and Uses for Objects Test performance added to the variance after controlling for verbal knowledge and speed. The results suggest that age-related decline in incidental recall is largely due to speed and the strategic search of memory. PMID- 11014712 TI - Cognitive complexity and cognitive-affective integration: related or separate domains of adult development? AB - This study used a latent variable approach to describe two broad domains of adult development and their interrelations. One domain was cognitive complexity, defined by crystallized intelligence, fluid intelligence, and reflective cognition: the other domain was cognitive-affective integration, operationalized in terms of integrated and defensive coping. It was hypothesized that these 2 domains are related to each other and that they imply different developmental correlates. Structural relations among the latent variables supported a model in which integrated coping showed positive relations with crystallized intelligence and reflective cognition, whereas defensive coping showed negative relations to these cognitive factors. Age and education were significant predictors of the cognitive complexity factors, whereas evaluations of the climate in one's current family and family of origin were related to the factors of cognitive-affective integration. PMID- 11014713 TI - Longitudinal changes in the contribution of genetic and environmental influences to symptoms of depression in older male twins. AB - Genetically informative longitudinal data on self-reported symptoms of depression allow for an investigation of the causes of stability and change in depression symptoms throughout adult life. In this report, the authors investigated the relative contribution of genetic and environmental influences to symptoms of depression in 83 monozygotic and 84 dizygotic male twin pairs from the National Heart, Lung, and Blood Institute (NHLBI) Twin Study. Participants first completed the Center for Epidemiologic Studies-Depression (CES-D) scale in 1985-1986 and again during 1995-1997. Mean age of twins at baseline was 63 years, range 59 to 70. From cross-sectional genetic analyses we estimated the heritability of CES-D to be 25% (95% confidence interval [CI], 11%-39%) at baseline and 55% (95% CI, 40%-71%) at follow-up. Fitting longitudinal genetic models to the two-wave data, we found that stability of symptoms over the 10-year follow-up was due primarily to continuity of genetic influences. PMID- 11014714 TI - Is age-related stability of subjective well-being a paradox? Cross-sectional and longitudinal evidence from the Berlin Aging Study. AB - Subjective well-being is thought to remain relatively stable into old age despite health-related losses. Age and functional health constraints were examined as predictors of individual differences and intraindividual change in subjective well-being, as indicated by positive and negative affect, using cross-sectional (N = 516) and longitudinal (N = 203) samples from the Berlin Aging Study (age range 70-103 years). In cross-sectional and longitudinal analyses, age and functional health constraints were negatively related to positive affect but unrelated to negative affect. Cross-sectionally, controlling for functional health constraints reversed the direction of the relationship between age and positive affect and produced a negative association between age and negative affect. Findings suggest two qualifications to the average stability of overall subjective well-being: Only some dimensions of subjective well-being remain stable, while others decline; age per se is not a cause of decline in subjective well-being but health constraints are. PMID- 11014715 TI - Aging and reflective processes of working memory: binding and test load deficits. AB - It was hypothesized that age-related deficits in episodic memory for feature combinations (e.g., B. L. Chalfonte & M. K. Johnson, 1996) signal, in part, decrements in the efficacy of reflective component processes (e.g., M. K. Johnson, 1992) that support the short-term maintenance and manipulation of information during encoding (e.g., F. 1. M. Craik. R. G. Morris. & M. L. Gick, 1990; T. A. Salthouse, 1990). Consistent with this, age-related binding deficits in a working memory task were found in 2 experiments. Evidence for an age-related test load deficit was also found: Older adults had greater difficulty than young adults when tested on 2 features rather than 1, even when binding was not required. Thus, disruption of source memory in older adults may involve deficits in both encoding processes (binding deficits) and monitoring processes (difficulty accessing multiple features, evaluating them, or both). PMID- 11014716 TI - Target selection difficulty, negative priming, and aging. AB - It has been recently suggested that the presence of identity negative priming effects in old adults could occur when there is substantial processing of the distracting information in a selective attention task (J. M. Kieley & A. A. Hartley, 1997). In three experiments, using a letter identification task, it was found that making target selection more difficult increased the magnitude of the negative priming effect to a similar extent in both young and old adults. Moreover, the size of the negative priming effect did not differ between young and elderly participants. These results are discussed with respect to the issue of age-related deficits in the mechanisms underlying negative priming. PMID- 11014717 TI - Instance-based automaticity and aging: acquisition, reacquisition, and long-term retention. AB - This research examined age differences in the acquisition and reacquisition of instance-based automaticity. In 2 experiments, young and older adults were trained to enumerate targets presented in otherwise empty displays or in displays that contained distractors. Experiment 1 revealed that older adults required more practice to reach asymptote than young adults. For both age groups, modifications of the identities and locations of targets produced substantial disruptions in performance, whereas modifications of the identities or locations of distractors produced little interference. However, no age differences in the representations of instances in memory were obtained in participants who reached asymptote. Experiment 2 revealed age deficits in the long-term retention and rate of reacquisition of instance-based automaticity 18 months after initial training. PMID- 11014718 TI - Dose-response evaluation of the therapeutic index for inhaled budesonide in patients with mild-to-moderate asthma. AB - PURPOSE: Inhaled corticosteroids have beneficial effects on pulmonary function and inflammation in patients with asthma, but they also cause systemic adverse effects, such as adrenal suppression. We evaluated the therapeutic index of inhaled corticosteroids in asthmatic patients by comparing their dose-response effects on lung function, surrogate markers of airway inflammation, and tests of adrenal function. SUBJECTS AND METHODS: After a 10-day placebo run-in, we evaluated the effects of 200 microg, 400 microg, and 800 microg of inhaled budesonide, each dose given twice daily sequentially for 3 weeks in 26 patients, aged 35 +/- 12 years (mean +/- SD), with mild-to-moderate asthma. Measurements were made of bronchial reactivity, exhaled nitric oxide (a marker of airway inflammation), spirometry, serum eosinophilic cationic protein concentration, and 10-hour overnight urinary cortisol excretion. Plasma cortisol levels were measured at 8 AM and after stimulation with human corticotropin releasing factor. RESULTS: For measurements of pulmonary function and exhaled nitric oxide, there was a plateau in the mean response to budesonide between 400 microg (low dose) and 800 microg (medium dose) per day, whereas for eosinophilic cationic protein and bronchial challenge, maximal benefits occurred between 800 and 1,600 microg (high dose) per day. Effects on plasma cortisol levels showed maximal suppression at 1,600 microg of budesonide per day. The proportion of patients with an optimal therapeutic index, in terms of a good airway response (fourfold decrease in bronchial hyperreactivity) and minimal systemic response (overnight urinary cortisol greater than 20 nmol), was similar at low-dose (46%) and at high-dose (52%) budesonide. The proportion of patients with a suboptimal therapeutic index, a good airway response with a marked systemic response (overnight urinary cortisol greater than 20 nmol), increased from 4% at low dose to 38% at high dose. CONCLUSIONS: In patients with mild-to-moderate atopic asthma, there were dose-related effects of budesonide on surrogate markers of inflammation (bronchial hyperreactivity and serum eosinophilic cationic protein), although higher doses were associated with adrenal suppression and a decrease in the therapeutic index. PMID- 11014719 TI - Ginkgo biloba extract for the treatment of intermittent claudication: a meta analysis of randomized trials. AB - PURPOSE: The optimal treatment of intermittent claudication has not yet been identified. Ginkgo biloba extract has been reported to have beneficial effects. We performed a meta-analysis of the efficacy of Ginkgo biloba extract for intermittent claudication based on the results of randomized, placebo-controlled, double-blind trials. METHODS: Literature searches of MEDLINE, EMBASE, BIOSIS, AMED, CISCOM, and the Cochrane Library were performed to identify studies on the topic. Manufacturers of commercial Ginkgo biloba products and authors of original publications and reviews were contacted to provide additional information. No language restrictions were imposed. RESULTS: Eight randomized, placebo controlled, double-blind trials were included. Meta-analysis found a significant difference in the increase in pain-free walking distance in favor of Ginkgo biloba (weighted mean difference: 34 meters, 95% confidence interval [CI]: 26 to 43 meters). In studies using similar methodological features (ergometer speed: 3 km/h, inclination: 12%) this difference was 33 meters in favor of Ginkgo biloba (95% CI: 22 to 43 meters). Adverse effects were rare, mild, and transient. CONCLUSIONS: These results suggest that Ginkgo biloba extract is superior to placebo in the symptomatic treatment of intermittent claudication. However, the size of the overall treatment effect is modest and of uncertain clinical relevance. PMID- 11014720 TI - A multicenter, randomized trial of fluconazole versus amphotericin B for empiric antifungal therapy of febrile neutropenic patients with cancer. AB - PURPOSE: To compare the efficacy and safety of fluconazole and amphotericin B as empiric antifungal therapy of febrile neutropenic patients with cancer. PATIENTS AND METHODS: A total of 317 neutropenic patients (<500 cells/mm3) with persistent or recrudescent fever despite 4 or more days of antibacterial therapy were randomly assigned to receive either fluconazole (400 mg intravenously once daily) or amphotericin B (0.5 mg/kg once daily). Patients were evaluated for the efficacy and safety of each drug by clinical criteria, frequent cultures and radiological procedures, and laboratory values. A response was classified as satisfactory at the end of therapy if the patient was afebrile, had no clinical or microbiological evidence of fungal infection, and did not require study termination due to lack of efficacy, drug toxicity, or death. RESULTS: A satisfactory response occurred in 68% of the patients treated with fluconazole (107 of 158 patients) and in 67% of patients treated with amphotericin B (106 of 159 patients). Progressive or new fungal infections during therapy occurred in 13 (8%) patients treated with fluconazole (8 with Candida, 5 with Aspergillus) and in 10 (6%) patients treated with amphotericin B (5 with Candida, 3 with Aspergillus, 2 with other fungi). Adverse events related to study drug (especially fever, chills, renal insufficiency, electrolyte disturbances, and respiratory distress) occurred more often in patients treated with amphotericin B (128 [81%] of 159 patients) than patients treated with fluconazole (20 [13%] of 158 patients, P = 0.001). Eleven (7%) patients treated with amphotericin B but only 1 (1%) patient treated with fluconazole were terminated from the study owing to an adverse event (P = 0.005). Overall mortality (27 [17%] patients treated with fluconazole versus 34 [21%] patients treated with amphotericin B) and mortality from fungal infection (7 [4%] patients treated with fluconazole versus 5 [3%] patients treated with amphotericin B) were similar in each study group. CONCLUSIONS: Intravenous fluconazole can be an effective and safe alternative to amphotericin B for empiric antifungal therapy in many febrile neutropenic patients. However, because fluconazole may be ineffective in the treatment of Aspergillus, patients at risk for that infection should be evaluated by chest radiograph, computed tomographic scanning, and cultures before the use of empiric fluconazole therapy. PMID- 11014721 TI - An outbreak of Legionella micdadei pneumonia in transplant patients: evaluation, molecular epidemiology, and control. AB - PURPOSE: To describe a nosocomial outbreak of Legionella micdadei pneumonia in transplant patients and to characterize the source of the outbreak and the control measures utilized. SUBJECTS AND METHODS: We performed retrospective Legionella micdadei serologic testing to enhance case finding in transplant patients with pneumonia that lacked a documented microbial etiology, as well as prospective environmental surveillance of water sites and testing for Legionella in clinical specimens. RESULTS: During a 3-month period, 12 cases of Legionella micdadei pneumonia were identified either by culture or serologic testing among 38 renal and cardiac transplant patients. Legionella micdadei isolates from hot water sources were found by pulsed-field gel electrophoresis to have a DNA banding pattern that was identical to the isolates from the first 3 culture positive cases and from 2 cases that occurred 16 months later. CONCLUSIONS: Hospitals caring for organ transplant recipients and other immunosuppressed patients must be aware of the possibility of environmental sources of outbreaks of Legionella infection. A first-line screen with the Legionella urine antigen test will identify Legionella pneumophila serogroup 1. However, specific cultures in outbreak situations should be considered to identify other Legionella pneumophila serotypes and the nonpneumophila Legionella species. PMID- 11014722 TI - Osteomalacia due to vitamin D depletion: a neglected consequence of intestinal malabsorption. AB - PURPOSE: Osteomalacia due to vitamin D depletion is believed to be rare in the United States because of the routine fortification of milk and other dairy products with vitamin D. We present a series of patients with histologically verified osteomalacia due to vitamin D depletion to emphasize the need for more careful and systematic surveillance of patients at risk of this metabolic bone disease. METHODS: Between 1989 and 1994, 17 patients with osteomalacia due to vitamin D depletion were seen in the Bone and Mineral Division of Henry Ford Health System, Detroit. All patients had a transiliac bone biopsy after in vivo double tetracycline labeling. Biochemical indexes of vitamin D nutritional status, parathyroid function, markers of bone turnover, and bone mineral density were assessed at the time of bone biopsy. The duration of symptoms, the lag between the cause of vitamin D depletion and the development of symptoms, and the radiologic findings were recorded. RESULTS: Osteomalacia was suspected by the referring physician in only 4 of the 17 patients, although a gastrointestinal disorder that can lead to vitamin D depletion was present in every patient. Thirteen of the patients had sustained at least one osteoporotic fracture (wrist, spine, or hip), and most had low appendicular and axial bone mineral density. All patients had one or more biochemical abnormalities consistent with vitamin D depletion. In 4 patients, a progressive rise in the serum alkaline phosphatase level was recorded but was not investigated until the patient presented with bone pain, muscle weakness, or fracture. CONCLUSIONS: Osteomalacia due to vitamin D depletion appears not to be suspected or diagnosed promptly in susceptible patients, perhaps because their physicians were not sufficiently aware of this condition. PMID- 11014723 TI - A meta-analysis of the Papanicolaou smear and wet mount for the diagnosis of vaginal trichomoniasis. AB - PURPOSE: To obtain reliable estimates of the sensitivity and specificity of the cervical Papanicolaou (Pap) smear and wet mount to diagnose vaginal trichomoniasis. METHODS: Articles indexed in MEDLINE (1976-1998) about diagnostic tests for trichomoniasis and their listed references were retrieved. Thirty studies (9,501 patients) that used trichomonas culture as a gold standard were selected. Studies were defined as level I if they fulfilled at least two of the following three criteria: consecutive patients were evaluated prospectively, the decision to culture was not influenced by test results, and there was independent and blind comparison with culture. Studies were classified as level II if one criterion was fulfilled, and as level III otherwise. RESULTS: The pooled sensitivity of the Pap smear for the diagnosis of trichomoniasis among level I studies was 57% (95% confidence interval [CI]: 51% to 63%) and the pooled specificity was 97% (95% CI: 93% to 100%). The likelihood ratio for a positive Pap smear was 19 among level I studies (range: 8 to 62). The pooled sensitivity of the wet mount among level I studies was 58% (95% CI: 51% to 66%); among level II studies, the sensitivity was 72% (95% CI: 62% to 81%), and among level III studies, the sensitivity was 82% (95% CI: 67% to 97%). The overall specificity of the wet mount was 99.8%. CONCLUSIONS: A positive Pap smear for trichomonads in settings in which trichomoniasis is common (prevalence > or =20%) requires treatment. A positive Pap smear is indeterminate when the prevalence of trichomoniasis is about 10%; thus, clinicians should either confirm the diagnosis by culture or treat all such patients, recognizing that some patients will be treated unnecessarily. A culture should be obtained in women with a positive Pap smear who are unlikely to have trichomoniasis (prevalence < or =1%). While a positive wet mount is diagnostic, a negative wet mount does not exclude trichomoniasis. PMID- 11014724 TI - Ischemic complications after percutaneous transluminal coronary angioplasty. AB - The ischemic complications ofpercutaneous transluminal coronary angioplasty (PTCA) include abrupt closure, which occurs in 2% to 10% of patients and is associated with increased morbidity and mortality. Periprocedural myocardial infarction due to side branch occlusion or embolization of platelet aggregates or thrombus occurs in 5% to 20% of patients. Patients with acute coronary syndromes, older age, and complex lesions are at greater risk of periprocedural complications. Technical advances, primarily stenting, are useful in the prevention and management of acute closure, but are also accompanied by thrombotic complications. It remains to be seen whether the new antithrombin agents reduce the rate of periprocedural complications if used in combination with aspirin and new antiplatelet therapies. These new antiplatelet agents (ticlopidine, clopidogrel, abciximab, eptifibatide, and tirofiban) reduce the rate of ischemic complications and have become standard adjunctive therapy for patients who undergo PTCA. PMID- 11014725 TI - Apoptosis and neurologic disease. AB - Many neurological disorders involve cell death. During development of the nervous system, cell death is a normal feature. Elimination of substantial numbers of initially generated cells enables useful pruning of "mismatched" or excessive cells produced by exuberance during the proliferative and migratory phases of development. Such cell death, occurring by "programmed" pathways, is termed apoptosis. In mature organisms, cells die in two major fashions, either by necrosis or apoptosis. In the adult nervous system, because there is little cell production during adulthood, there is little normal cell death. However, neurological disease is often associated with significant neural cell death. Acute disorders, occurring over minutes to hours, such as brain trauma, infarction, hemorrhage, or infection, prominently involve cell death, much of which is by necrosis. Chronic disorders, with relatively slow central nervous system degeneration, may occur over years or decades, but may involve cell losses. Such disorders include motor neuron diseases such as amyotrophic lateral sclerosis (ALS), cerebral dementing disorders such as Alzheimer's disease and frontotemporal dementia, and a variety of degenerative movement disorders including Parkinson's disease, Huntington's disease, and the inherited ataxias. There is evidence that the mechanism of neuronal cell death in these disorders may involve apoptosis. Direct conclusive evidence of apoptosis is scarce in these chronic disorders, because of the swiftness of cell death in relation to the slowness of the disease. Thus, at any particular time point of assessment, very few cells would be expected to be undergoing death. However, it is clearly of importance to define the type of cell death in these disorders. Of significance is that while treating the underlying causes of these conditions is an admirable goal, it may also be possible to develop productive therapies based on alleviating the process of cell death. This is particularly likely if this cell loss is through apoptosis, a programmed process for which the molecular cascade is increasingly understood. This article reviews our understanding of apoptosis in the nervous system, concentrating on its possible roles in chronic neurodegenerative disorders. PMID- 11014726 TI - Accuracy and cost-effectiveness of single-view echocardiographic screening for suspected mitral valve prolapse. PMID- 11014727 TI - Supine exercise capacity identifies patients at low risk for perioperative cardiovascular events and predicts long-term survival. PMID- 11014728 TI - Inhaled corticosteroid therapy for asthma: therapeutic and toxic potentials. PMID- 11014729 TI - Ginkgo biloba: a living fossil. PMID- 11014730 TI - Management of persistent fever in the neutropenic patient. PMID- 11014731 TI - Nosocomial Legionella micdadei infection in transplant patients: fortune favors the prepared mind. PMID- 11014732 TI - Increased use of beta-blockers after acute myocardial infarction. PMID- 11014733 TI - Hepatitis C and B-cell non-Hodgkin's lymphoma: a geographically variable association? PMID- 11014734 TI - Fatal rhabdomyolysis associated with simvastatin in a renal transplant patient. PMID- 11014735 TI - Fatal ventricular fibrillation during a low-dose dobutamine stress test. PMID- 11014737 TI - Managed care and education: seizing the opportunities. PMID- 11014736 TI - Minocycline-induced systemic adverse reaction with liver and bone marrow granulomas and Sezary-like cells. PMID- 11014738 TI - The past as prologue to the future: the times, they've been a-changin'. PMID- 11014739 TI - Developmental psychopathology: concepts and challenges. AB - The defining features of developmental psychopathology concepts include attention to the understanding of causal processes, appreciation of the role of developmental mechanisms, and consideration of continuities and discontinuities between normality and psychopathology. Accomplishments with respect to these issues are reviewed in relation to attachment disorders, antisocial behavior, autism, depressive disorder, schizophrenia, and intellectual development. Major research challenges remain in relation to measurement issues, comorbidity, gender differences, cognitive processing, nature-nurture interplay, heterotypic continuity, continuities between normal variations and disorders, developmental programming, and therapeutic mechanisms in effective treatments. PMID- 11014740 TI - Developmental systems and psychopathology. AB - Efforts to understand the etiology of adult mental disorders by studying children has produced unanticipated changes in our understanding of pathology, individual development, and the role of social context. Among these are the blurring of the division between mental illness and mental health, the need to attend to patterns of adaptation rather than personality traits, and the powerful influences of the social world on individual development. Current developmental views place deviancy in the dynamic relation between individuals and their contexts. At another level, when we view the history of developmental psychopathology, dialectical developmental processes are evident as we trace how patterns of adaptation of researchers, expressed in theoretical models and empirical paradigms. increasingly have come to match the complexities of human mental health and illness. PMID- 11014741 TI - Experience in the midst of variation: new horizons for development and psychopathology. AB - This essay explores implications of current trends in developmental science for understanding psychopathology at the dawn of the new millennium. Over the past half century, it has become clear that uniform and general principles of development (i.e., those that are applicable at all times, to all people, and in all places) will be of limited utility in understanding the processes of greatest interest in development and psychopathology. Instead, such processes are characterized by complexly organized individuals engaged in developmental transactions within multiple contexts (ranging from the biological environment of neurons to the cultural systems of meaning that shape people's lives). These transactions in turn often yield variable outcomes. In order to portray how we have come to this conclusion, we first provide a view of contemporary research in three areas of early development: the biology of the developing brain, the complexities of early emotional development, and the cultural contexts of child development. We then trace how an increasing appreciation of organized complexity, developmental transactions, and the meaning of context have played out in the emerging field of infant mental health before closing with our vision of new opportunities for the study of experience in the midst of variation. PMID- 11014742 TI - Cultural influences on developmental processes and outcomes: implications for the study of development and psychopathology. AB - The purpose of this paper is to trace the role of culture as an explanatory construct in developmental processes and outcomes, and its implications in the understanding of developmental psychopathology. Literature reviews were conducted by historical period: 1930-1939, 1960-1969, and 1990-1999. The percentage of the total articles and chapters pertaining to cultural issues increased as a function of time. Both conceptual and methodological continuities and discontinuities were observed among the three periods. The preponderance of comparative studies using deficit models still remains, but more enlightened alternative conceptual models, within culture studies, and measures of cultural processes, are emerging. In contrast, although contextual influences are considered important in developmental psychopathology, the field lags in its empirical consideration of cultural influences. The need to seriously address these issues will increase as globalization and rapid cultural change become even more the norm than the exception. PMID- 11014744 TI - Psychosocial influences: critiques, findings, and research needs. AB - Nongenetic factors have a major influence on psychopathology. Knowledge on specific psychosocial risk and protective mechanisms is more limited because of inadequate attention to measurement issues, person effects on the environment, and the possibility of genetic mediation. Nevertheless, a range of research strategies may be used to provide rigorous tests of causal hypotheses; these have shown the importance of environmentally mediated risks. Challenges for the future include greater use of such research strategies, improved measures of psychosocial risks that can be applied to large samples, investigation of origins of risks, identification of causes of time trends in levels of psychopathology, delineation of psychosocial effects on lifetime liability, understanding of environmental effects on the organism, appreciation of processes involved in developmental programming, and understanding of individual differences in susceptibility. PMID- 11014743 TI - The interplay of genetic influences and social processes in developmental theory: specific mechanisms are coming into view. AB - In the coming years we can look forward to research that clarifies specific mechanisms that account for the interplay between genetic and environmental influences on psychological development. Certain misconceptions, arising from research traditions initiated by Francis Galton on the one hand and G. Stanley Hall on the other, may now be set aside in the light of new evidence. Three important findings promise a new synthesis. First, while each of us is born with about 100,000 genes that, under ordinary circumstances, do not change, the expression of these genes on behavior is dynamic. Some genetic influences are expressed early in development, but others are manifest many years later. Second, genetic factors often account not only for some of the individual differences in the measures of adjustments we typically use to monitor development but also for individual differences in environmental experiences that covary with those measures of adjustment. Indeed, genetic factors have been found to account for a surprising amount of covariance between measures of the social environment and of adjustment in young children, adolescents, and adults. Third, the expression of genetic influences are very malleable and responsive to the social environment. These new findings are revealing specific mechanisms for the interplay of genetic and social environmental factors in four domains. First, the social environment may play both a necessary and specific role in the expression of particular genetic influences on a range of behaviors from depression to social responsibility. Second, an understanding of the interplay between the social environment and genetics may lead to a clearer definition of the phenotypic manifestations of particular genetic influences. Third, we will-as a result of these studies-have a clearer fix on the timing of important events and their sequence in development. Fourth, this new genre of work promises to illumine more completely mechanisms by which the social environment influences development independent of genetic influence. PMID- 11014745 TI - The place of psychodynamic theory in developmental psychopathology. AB - Psychoanalysis ushered in this century. Will its influence on developmental psychopathology end in the next? The paper explores some critical obstacles in the way of psychodynamic research, including the fragmentation of psychoanalytic theory. the relative independence of theory from its clinical and empirical base, the predominance of inductive scientific logic, the polymorphous use of terms, the privacy of clinical data, the dominance of the reconstructionist stance, and the isolation of psychoanalysis from psychology and neurobiology. Notwithstanding these limitations, core psychoanalytic precepts are not only consistent with some of the most important advances of the last decade but may also be helpful in elaborating these new discoveries in the next century. Psychoanalysis is centered on the notion that complex, conflicting, unconscious representations of mental states constitute a key facet of normal and abnormal development. This notion retains its power, and deserves a prominent position among the major frames of reference to guide developmental science in the next century. PMID- 11014746 TI - Developing mechanisms of self-regulation. AB - Child development involves both reactive and self-regulatory mechanisms that children develop in conjunction with social norms. A half-century of research has uncovered aspects of the physical basis of attentional networks that produce regulation, and has given us some knowledge of how the social environment may alter them. In this paper, we discuss six forms of developmental plasticity related to aspects of attention. We then focus on effortful or executive aspects of attention, reviewing research on temperamental individual differences and important pathways to normal and pathological development. Pathologies of development may arise when regulatory and reactive systems fail to reach the balance that allows for both self-expression and socially acceptable behavior. It remains a challenge for our society during the next millennium to obtain the information necessary to design systems that allow a successful balance to be realized by the largest possible number of children. PMID- 11014747 TI - Internalizing problems of childhood and adolescence: prospects, pitfalls, and progress in understanding the development of anxiety and depression. AB - The focus of this article is on internalizing problems that are experienced by children and adolescents. We provide an historical perspective, selectively examine the current state of knowledge, consider advances and gaps in what is known, and identify new research directions. Diagnosis, epidemiology, theory, and research first are considered separately for anxiety and depressive disorders. These internalizing problems, however, whether clinical or subclinical, share many common features and show high comorbidity rates. We emphasize the importance of systematic analysis of comorbid anxiety and depression, including their comorbidity with externalizing problems. This could lead to more valid classification of subtypes of internalizing problems and further an understanding of the diverse conditions that constitute internalized distress. We highlight the need to study anxiety and depression within a developmental psychopathology framework, as well as to include both categorical and dimensional assessments of these problems in the same research designs. This will be essential for understanding the complex interplay of biological and environmental processes that contribute to the emergence, progression, and amelioration of internalizing problems over time. PMID- 11014748 TI - Early externalizing behavior problems: toddlers and preschoolers at risk for later maladjustment. AB - The early emergence and developmental implications of externalizing behavior problems in toddlers and preschoolers are discussed with an emphasis on which young children are truly at risk for continuing problems. The extant literature is reviewed with a focus on the stability of early externalizing behavior and the diverse pathways that young children, primarily boys, with early-emerging problems may follow. Findings from a number of studies, both epidemiological and high risk, suggest that the small subgroup of boys with multiple risk factors that include especially high levels of early hyperactivity and aggression, and high levels of negative parenting and family stress, are most likely to evidence continuing problems at school entry. Sociodemographic and neighborhood influences are also discussed, as are implications for future research and policy. PMID- 11014749 TI - Is asperger syndrome/high-functioning autism necessarily a disability? AB - This article considers whether Asperger syndrome (AS) or high-functioning autism (HFA) necessarily leads to disability or whether AS/HFA simply leads to "difference." It concludes that the term "difference" in relation to AS/ HFA is a more neutral, value-free, and fairer description than terms such as "impairment," "deficiency," or "disability"; that the term "disability" only applies to the lower functioning cases of autism; but that the term "disability" may need to be retained for ASIHFA as long as the legal framework provides financial and other support only for individuals with a disability. Two models are summarized which attempt to define in what way individuals with AS/HFA are "different": the central coherence model, and the folk psychology-folk physics model. The challenge for research is to test the value of such models and to precisely characterize the differences in cognitive style. PMID- 11014750 TI - The neurodevelopmental hypothesis of schizophrenia: following a trail of evidence from cradle to grave. AB - This is a critical review of the literature related to the neurodevelopmental hypothesis of schizophrenia which posits that the illness is related to abnormal brain development. The review focuses on data deriving from clinical studies, and it is organized according to the life phase from which the data were collected: conception and birth, infancy and childhood up to the onset of the illness, after illness onset, and postmortem. The neurodevelopmental hypothesis is supported by several pieces of evidence, including increased frequency of obstetric complications in patients with schizophrenia: the presence of minor physical anomalies; the presence of neurological, cognitive, and behavioral dysfunction long before illness onset; a course and outcome of the illness itself that is incompatible in most cases with a degenerative illness; the stability of brain structural measures over time; and the absence of postmortem evidence of neurodegeneration. A historical perspective on how this research accumulated and a section addressing important areas of future investigation are also provided. We conclude that schizophrenia is certainly not a degenerative brain disorder, and that it is likely that a brain insult in utero or at birth plays a role in its expression. Current evidence cannot completely exclude the role of environmental variables after birth. In addition, it is possible that other psychiatric disorders may also have a neurodevelopmental component. PMID- 11014751 TI - Integrating competence and psychopathology: pathways toward a comprehensive science of adaptation in development. AB - This paper examines the conceptual and empirical connections between competence and psychopathology, two historically rich traditions for the study of adaptation in development, and what might be gained from their integration. Historical roots of these two traditions are reviewed, then overlaps in their definition are considered, with a focus on the ways in which judgments about competence enter into the nosology of mental disorders. DSM-IV is analyzed from the perspective of competence, and the debate about "harmful dysfunction" in defining mental disorder is discussed in relation to competence. Different models explaining the empirical associations of competence and psychopathology are delineated, and illustrative empirical evidence is provided. Potential explanations include confounded concepts and methods, symptoms undermining the effectiveness of adaptation in the environment, failures in age-salient developmental tasks leading to emotional and behavioral problems, transactional influences, shared vulnerability or risk factors producing both kinds of difficulties, and more complex models. The potential benefits of integrating competence and psychopathology as two major approaches to adaptation are discussed in regard to theory, classification of mental disorder, research, and intervention. PMID- 11014752 TI - Behavioural implications of alarm mistrust as a function of task workload. AB - The research was conducted to investigate the effect of increasing primary task and alarm workload on alarm mistrust as reflected by alarm and primary task performances. A total of 126 undergraduate students performed a complex psychomotor task battery three times, with the number of concurrent tasks increasing each time. During their performance, the students were required to react to an alarm system (including visual and auditory components) of questionable reliability. Depending on the group to which participants were assigned, the alarm presentation rate constituted a low-, medium- or high workload condition. Alarm response data (times, frequencies, accuracies) and primary task data (tracking error) were analyzed to assess performance differences as a function of primary and secondary task workload levels. Results generally supported the hypotheses: increasing primary task and alarm task workload degraded alarm response performance. Also, response frequencies supported earlier research suggesting that participants 'probability match' their response rates to alarm system reliability. The results are discussed with regard to the cry-wolf effect, attention theory and alarm system design. PMID- 11014753 TI - Analysis of 3-D human foot forms using the Free Form Deformation method and its application in grading shoe lasts. AB - An effective way to design well-fitting products is to analyse human body forms and to classify them into several groups. In the present study, a new method is proposed to analyse human body forms using the FFD (Free Form Deformation) technique. The FFD method is a way to deform the shapes of object smoothly by moving control lattice points set around the object. The reference body form is automatically deformed to coincide with the other body forms using the FFD method. The dissimilarity is defined by the movements of the control lattice points. The foot forms of 56 Japanese adult females were analysed with this method, and distributions for them were calculated using multi-dimensional scaling. The first axis contrasts feet with high dorsal arches and low dorsal arches, and the second axis is related to the antero-posterior proportion of the foot. As an application of the present method, a last of width EEEE was designed from an existing last of width E by applying the control lattice points that converted a representative foot of width E into a foot of width EEEE. The new EEEE width last reflected the allometric differences between narrow and wide feet better than one obtained by a conventional method. It was found that the present method with FFD is not only useful for classifying 3-D human body forms, but also has potential as applications for designing well-fitting products. PMID- 11014754 TI - A three-dimensional dynamic posture prediction model for simulating in-vehicle seated reaching movements: development and validation. AB - A three-dimensional dynamic posture prediction model for simulating in-vehicle seated reaching movements is presented. The model employs a four-segment 7 degrees-of-freedom linkage structure to represent the torso, clavicle and right upper extremity. It relies on an optimization-based differential inverse kinematics approach to estimate a set of four weighting parameters that quantify a time-constant, inter-segment motion apportionment strategy. In the development phase, 100 seated reaching movements performed by 10 subjects towards five typical in-vehicle targets were modelled, resulting in 100 sets of weighting parameters. Statistical analysis was then conducted to relate these parameters to target and individual attributes. In the validation phase, the generalized model, with parameter values statistically synthesized, was applied to novel data sets containing 700 different reaching movements (towards different targets and/or by different subjects). The results demonstrated the model's ability to generate close representations in prediction: the overall mean time-averaged error in joint angle was 5.2 degrees, and the median was 4.7 degrees, excluding reaches towards two extreme targets (for which modelling errors were excessive). The model's general success in prediction and its unique characteristics led to implications with regard to the performance and underlying control strategies of human reaching movements. PMID- 11014755 TI - Kinetic changes associated with load carriage using two rucksack designs. AB - This study assessed changes in kinetics from unloaded walking associated with load carriage using both a traditional and a new rucksack design that incorporates front balance pockets (AARN). Nine subjects walked at 3(+/-0.05) km x h(-1) over a force plate unloaded and carrying 25.6 kg in each of the rucksacks. The order of trials was randomized and speed-controlled by use of photoelectric cells and a millisecond timer. Anteroposterior and vertical ground reaction forces were analyzed using repeated measures ANCOVA (speed covariate). There was a trend for the AARN pack to elicit a shorter support time than the traditional pack, 1.025+/-0.049 versus 1.037+/-0.06 s (p = 0.056), while the unloaded condition produced the shortest support time, 1.016+/-0.04 s. Both braking and propulsive forces for the rucksacks were significantly greater than for unloaded walking. While there was no significant difference between the packs for the braking force, the AARN pack produced a significantly lower (p<0.05) propulsive force than the traditional rucksack, 0.79+/-0.2 versus 0.94+/-0.16 N x kg bodyweight(-1). Both rucksacks produced significantly greater (p<0.001) vertical force peaks than unloaded walking, the increases being proportional to the increase in system weight. These findings indicate that there may be some advantage in terms of propulsive force production for the front/back system. PMID- 11014756 TI - After-effects of the common cold on mood and performance. AB - The present study examined whether volunteers who had recently had common colds showed impairments in mood and performance in the weeks following the illness. All volunteers (n = 24) were tested when healthy to provide baseline data for simple and choice reaction time tasks, attention and memory tasks and ratings of mood. When participants developed a cold (n = 13) they returned to the laboratory so that the illness could be verified. When they were symptom free they returned to the laboratory and repeated the procedure. They then completed the study with a final session 1 week later. Volunteers (n = 11) who remained healthy over 10 weeks were recalled as controls and also repeated the procedures. The results showed that those who had recently had colds showed few impairments in mental performance and mood. Taken together with the results of previous studies, this suggests that after-effects of viral infection are largely restricted to severe illnesses such as infectious mononucleosis and influenza. After-effects of colds may occur but these probably reflect poor learning at the time of the illness. PMID- 11014757 TI - Cardiorespiratory and thermoregulatory response of working in fire-fighter protective clothing in a temperate environment. AB - The cardiorespiratory and thermal responses of two intensities of treadmill exercise were compared for brief periods (12 min) in fire ensemble (FE) but without self contained breathing apparatus, and sports ensemble (SE), in a temperature environment. A further experiment explored the responses of subjects exercising in FE over a prolonged period (60 min). Eighteen male fire-fighters wearing either FE or SE walked on a level treadmill for 6 min at 5 km x h(-1) increasing to 7 km x h(-1) for 6 min. Following a recovery interval of 1 h, the exercise protocol was repeated in the second ensemble; the order of ensemble was balanced. Heart rate (HR), rectal temperature (Tre), VO2 max and rating of perceived exertion (RPE) were monitored continuously under both ensembles. At 7 km x h(-1), VO2 was significantly higher (p<0.05) in FE (36.1 and 39.9 ml x kg( 1) x min(-1)) than in SE and represented 74% VO2 max. There were no changes Tre. In experiment 2, following a rest interval of at least 36 h, eight subjects in FE walked on the treadmill at 6 km x h (gradient 10%) for 60 min also in temperate conditions, where HR, Tre and RPE were recorded at 10-min intervals. During the 60-min exercise in FE, HR reached 161 beats x min(-1) and Tre increased to 38.3 degrees C. Despite considerable subject discomfort, Tre remained below dangerous levels (38.4 degrees C). When RPE were compared with a physiological strain index (PSI) calculated from Tre and HR data over 60 min, there was no significant difference (p<0.05) with a correlation coefficient (r) of 0.98. The results suggest that RPE and PSI are closely related when exercise is sufficiently prolonged or intense to elevate Tre and HR in fire-fighters wearing FE in temperate conditions. If further investigation confirms this relationship for hot humid conditions in which fire-fighters operate, then with training, it may provide individuals with a valid measure of dangerous levels of perceived heat strain. PMID- 11014758 TI - Behavioural compensation by drivers of a simulator when using a vision enhancement system. AB - Technological progress is suggesting dramatic changes to the tasks of the driver, with the general aim of making driving environment safer. Before any of these technologies are implemented, empirical research is required to establish if these devices do, in fact, bring about the anticipated improvements. Initially, at least, simulated driving environments offer a means of conducting this research. The study reported here concentrates on the application of a vision enhancement (VE) system within the risk homeostasis paradigm. It was anticipated, in line with risk homeostasis theory, that drivers would compensate for the reduction in risk by increasing speed. The results support the hypothesis although, after a simulated failure of the VE system, drivers did reduce their speed due to reduced confidence in the reliability of the system. PMID- 11014759 TI - Fault management in supervisory control: the effect of false alarms and support. AB - Automation has changed the role of human operators from direct manual control to supervision. Their main task is to monitor whether system performance remains within pre-specified ranges and intervention is only required in unusual situations. One of the consequences is a loss of situation awareness, which significantly affects performance in abnormal, time-critical situation. The present study reports two experiments, both dealing with fault management in a maritime supervisory control task. The first experiment investigated to what extent false alarms would affect performance and diagnosis behaviour when multiple disturbances occurred. Thirty-nine students from maritime curricula diagnosed disturbances that could either be real or turn out to be a false alarm. The presence of false alarms not only affected the rate with which the subsystems under control were sampled, but it also increased problem-solving time. One of the reasons for suboptimal performance in dealing with fault propagation was tunnel vision: participants had a tendency to deal with disturbances sequentially. In the second experiment the effect of support on performance and diagnosis behaviour was investigated. Two types of support were distinguished: interactive support requiring participants to provide the symptom values and automatic support that directly provided the correct action. Thirty students from maritime curricula diagnosed disturbances with the help of either the interactive or the noninteractive support tool. The results indicated that even though both support tools gave the same advice on how to act, more incorrect actions were taken in the non-interactive support condition. Even though no differences in performance were found after the tool had been removed, it was shown that participants who were used to interactive support used a more structured problem solving strategy than participants used to the non-interactive support. Consequences for system design are discussed. PMID- 11014760 TI - Maximum reach envelope for the seated and standing male and female for industrial workstation design. AB - Maximum reach envelopes for the 5th, 50th and 95th percentile reach lengths of males and females in seated and standing work positions were determined. The use of a computerized potentiometric measurement system permitted functional reach measurement in 15 min for each subject. The measurement system captured reach endpoints in a dynamic mode while the subjects were describing their maximum reach envelopes. An unbiased estimate of the true reach distances was made through a systematic computerized data averaging process. The maximum reach envelope for the standing position was significantly (p<0.05) larger than the corresponding measure in the seated position for both the males and females. The average reach length of the female was 13.5% smaller than that for the corresponding male. Potential applications of this research include designs of industrial workstations, equipment, tools and products. PMID- 11014762 TI - Use of accelerometers as an ergonomic assessment method for arm acceleration-a large-scale field trial. AB - Ergonomists need easy-to-use, quantitative job evaluation methods to assess risk factors for upper extremity work-related musculoskeletal disorders in field-based epidemiology studies. One device that may provide an objective measure of exposure to arm acceleration is a wrist-worn accelerometer or activity monitor. A field trial was conducted to evaluate the performance of a single-axis accelerometer using an industrial population (n=158) known to have diverse upper limb motion characteristics. The second phase of the field trial involved an examination of the relationship between more traditional observation-based ergonomic exposure measures and the monitor output among a group of assembly-line production employees (n=48) performing work tasks with highly stereotypic upper limb motion patterns. As expected, the linear acceleration data obtained from the activity monitor showed statistically significant differences between three occupational groups known observationally to have different upper limb motion requirements. Among the assembly-line production employees who performed different short-cycle assembly work tasks, statistically significant differences were also observed. Several observation-based ergonomic exposure measures were found to explain differences in the acceleration measure among the production employees who performed different jobs: hand and arm motion speed, use of the hand as a hammer, and, negatively, resisting forearm rotation from the torque of a power tool. The activity monitors were found to be easy to use and non intrusive, and to be able to distinguish arm acceleration among groups with diverse upper limb motion characteristics as well as between different assembly job tasks where arm monitors were performed repeatedly at a fixed rate. PMID- 11014761 TI - Experimental investigation and biomechanical analysis of lower limb movements for clutch pedal operation. AB - Lower limb movements for clutch-pedal operations were investigated and the influence of four parameters (seat height, pedal travel, pedal travel inclination, pedal resistance) was studied using a multi-adjustable experimental seat. Fifteen subjects participated in the experiment: five short females, five average height males and five tall males. A biomechanical model has been proposed to explain how pedal force direction could be controlled. The experimental observations show that the lower limb movement of clutch pedal operation is mainly guided by the geometric constraints imposed by the task and its environment, especially during the depression phase. The results support the hypothesis that movements obey the principle of minimum work and minimum discomfort. Furthermore, it seems that a functional segmentation exists between the distal joint (ankle) and the proximal joints (knee and hip), thus simplifying the control problem, which is due to the redundancy of the human body. It appears that the depression movement is controlled by proprioceptive feedback related to foot displacement and pedal force, from the fact that the deceleration duration during the depression phase increases with the pedal resistance and pedal travel. The minimum pedal resistance and pedal travel are discussed. PMID- 11014763 TI - Transition from Journal of Diarrhoeal Diseases Research to Journal of Health, Population and Nutrition. PMID- 11014764 TI - Rotavirus serotypes: classification and importance in epidemiology, immunity, and vaccine development. AB - The development and implementation of safe and effective vaccines to prevent the enormous health burden of rotavirus-associated disease is a global public health goal. Human rotaviruses, the major aetiological agents of severe infantile diarrhoea worldwide, display surprisingly diverse and complex serotypic specificities. Ten VP7 serotypes and 7 VP4 serotypes have so far been detected. An increasing number of observations, obtained from analyses of (i) natural rotavirus infections in infants and young children, (ii) experimental rotavirus infections in laboratory animals, and (iii) extensive rotavirus vaccine field trials performed in different populations of various parts of the world, appears to support the concept that serotype-specific antibodies to rotaviruses play an important role in protection against rotavirus-associated illnesses. Thus, the first licensed rotavirus vaccine (RRV-based quadrivalent vaccine) was designed to cover the epidemiologically important VP7 serotype 1, 2, 3, and 4. PMID- 11014765 TI - Prevention of diarrhoea in a poor District of Santo Domingo, Dominican Republic: practices, knowledge, and barriers. AB - The study, conducted in a poor periurban community of Santo Domingo, Dominican Republic, assessed the practices, knowledge, and barriers relating to prevention of diarrhoea. A total of 582 caregivers of children, aged less than 5 years, were systematically sampled from four barrios. Results of the study showed that 55% of the caregivers did not boil drinking water for children; 38% did not always wash hands of the children prior to meals; 87% of the children did not always wear shoes outside their house; and 54% were breastfed for less than one year. Biomedical knowledge about these practices was high among the caregivers, and was not related to the reported behaviours. However, several barriers were significantly related to practices, including lapse in caregiving, limited resources, erroneous beliefs, and non-compliance by children. Health education, based on a biomedical knowledge-deficit model, may have little impact on improving the diarrhoea-prevention practices in these communities. Greater attention should, therefore, be directed toward the barriers experienced by caregivers of children. PMID- 11014766 TI - Identification of enteric pathogens in HIV-positive patients with diarrhoea in northern India. AB - Enteric pathogens associated with chronic diarrhoea in HIV-positive patients were studied. The study was conducted during January 1995-December 1998. Stool specimens from all diarrhoea patients (n = 26) were examined microscopically for ova and parasites using wet preparations and stained smears. Stool samples from diarrhoea patients were also cultured on appropriate media to isolate enteric bacterial pathogens. Of the 59 patients, 26 (44%) had prolonged diarrhoea for more than 4 weeks. Enteric pathogens were detected in 19 (73%) of the 26 patients: 17 patients harboured a single pathogen, and 2 patients had mixed pathogens. The detection rate of emerging parasites, including Isospora, Cryptosporidium, Blastocystis hominis, and Strongyloides stercoralis as a single agent, was significantly higher than conventional pathogens (50% vs 19.2%; p < 0.05). Only one patient harboured both conventional and emerging pathogens (Entamoeba histolytica and Cryptosporidium). Isospora belli was detected in 8 (31%) of the 26 diarrhoea patients: in 7 (27%) patients as a single agent and in one patient with S. stercoralis. Cryptosporidium was identified in 3 (11%) diarrhoea patients: in 2 (8%) patients as a single agent and in one patient with E. histolytica, followed by B. hominis in 2 (8%) patients. E. histolytica was most commonly isolated (3/26; 11.5%), followed by Giardia lamblia, enteropathogenic Escherichia coli, and Campylobacter jejuni (one patient each). Parasitic pathogens were frequently associated with HIV-positive patients with diarrhoea in northern India. I. belli was the most frequent parasite isolated, followed by Cryptosporidium. Stools of all HIV-positive patients with diarrhoea should thoroughly be investigated to identify aetiologic agents for proper management. PMID- 11014767 TI - Vibrio cholerae O139 Bengal: a descriptive study. AB - A prospective study was conducted to determine the clinical and laboratory characteristics and the clinical course of cholera due to Vibrio cholerae O139 Bengal. The study subjects included 22 adult males with stool culture-proven V. cholerae O139. On enrollment, mean +/- SD concentrations (mmol/L) of serum sodium, potassium, chloride, and bicarbonate were 134 +/- 3, 4 +/- 1, 102 +/- 4, and 13 +/- 4 respectively, and stool sodium, potassium, chloride, and bicarbonate concentrations were 120 +/- 24, 18 +/- 6, 93 +/- 16, and 37 +/- 9 respectively. Seventeen patients (7.8%) had faecal leukocytes ranging from 11 to 50 per high power field. All V. cholerae O139 isolates (100%) were susceptible to tetracycline, erythromycin, and ciprofloxacin, 92% to furazolidine, and only 5% to trimethoprim-sulphamethaxazole. The median (interquartile) volume of liquid stool during the first 24 hours was 9 (5-12) litre. The median (interquartile) volume of liquid stool and the amounts of intravenous and oral rehydration fluids required during the entire study period were 16 (9-24) litre, 9 (6-18) litre, and 14 (9-20) litre respectively. The median (interquartile) duration of diarrhoea was 80 (48-104) hours. The median (interquartile) duration of excretion of V. cholerae O139 in stool was 5 (3-6) days. Clinical and laboratory features, and case management of cholera due to V. cholerae O139 are very similar to conventional cholera due to V. cholerae O1. PMID- 11014768 TI - Isolation and antibiotic susceptibility of Salmonella, Shigella, and Campylobacter from acute enteric infections in Egypt. AB - While Campylobacter, Salmonella, and Shigella remain major contributors to acute enteric infections, few studies on these pathogens have been conducted in Egypt. From January 1986 to December 1993, 869 Salmonella, Shigella and Campylobacter strains were isolated from stool specimens from 6,278 patients, presenting to the Abbassia Fever Hospital, Cairo, Egypt, with acute enteric infections. Salmonella predominated, totalling 465 isolates, followed by Shigella with 258 isolates, and Campylobacter with 146 isolates. Of the Shigella isolates, 124 were Shigella flexneri, 49 were S. sonnei, 47 were S. dysenteriae (mainly serotype 1, 2, and 3), and 38 were S. boydii. Campylobacter spp. comprised 92 Campylobacter jejuni and 54 C. coli isolates. Isolation of Salmonella was highest during the months of February-March, June-July, and October-November, while that of Shigella was maximal from July to October. Isolation of Campylobacter increased during May June and again during August-October. Although Salmonella was sensitive to amikacin, aztreonam, ceftriaxone, and nalidixic acid, it was, however, resistant to erythromycin, streptomycin, ampicillin, chloramphenicol, and tetracycline. Shigella (> 80%) was sensitive to amikacin, ceftriaxone, cephalothin, sulphamethoxazole-trimethoprim (except S. sonnei), aztreonam, and nalidixic acid. Resistance (> 50%) was noted only for ampicillin, chloramphenicol, and tetracycline. C. jejuni and C. coli were resistant to cephalothin, aztreonam, and streptomycin. Some of the above antibiotics were employed to characterize the Egyptian isolates, but did not have any clinical utility in the treatment of diarrhoea. Significant differences (p < 0.05) were observed in the resistance profiles of Shigella and Salmonella between late 1980s and early 1990s. The results suggest the use of fluoroquinolones or a third-generation cephalosporin as an empirical treatment of enteric diseases. However, alternative control strategies, including the aggressive development of broadly protective vaccines, may be more effective approaches to curbing morbidity and mortality due to acute enteric infections. PMID- 11014769 TI - Serotypes and subgroups of rotavirus isolated from children in central Brazil. AB - Group A rotavirus, obtained from children of Goiania, Brazil, during 1987-1994, were analyzed for subgroup and G serotype by enzyme-linked immunosorbent assay with monoclonal antibodies. The index of serotyping obtained was 61.4% with the following proportions: G1--19.7%, G2--28.0%, G3--9.8%, G4--1.5%, and G5--2.3%. It was observed that G1 occurred from 1987 to 1989 and from 1993 to 1994, and G2 from 1990 to 1993. About 94% of the samples (85/90) could be subgrouped with the following results: 55.5% for SG II, 7.8% SG I, and 31.1% for SG non-I-non-II. Unusual relationship patterns were also detected among serotypes, subgroups, and profiles of electropherotypes in 57.0% of the samples: 20 of them were G2/SG II/"long" profile. The results suggest that variation in temporal and regional characteristics should be considered in the development of rotavirus vaccine. PMID- 11014770 TI - Production of heat-labile enterotoxin by strains of Aeromonas veronii bv veronii. AB - Three isolates of Aeromonas veronii bv veronii (2 environmental, one blood infection) were examined to see if they produce any enterotoxin and, if so, to determine its characteristics. Two isolates caused fluid accumulation in the initial rabbit ileal loop tests. The other strain did so after a single passage through the rabbit ileal loop. All the isolates showed gradual enhancement of fluid secretion after each subsequent passage. Inocula of 1 x 10(4) viable cells and 0.25 mL of culture filtrate caused fluid accumulation comparable to those of toxigenic Vibrio cholerae 569B. The enterotoxic activity was inactivated at higher temperature, and showed biological activity over a wide range of pH. The only histopathological change observed was depletion of mucous from goblet cells. The findings of the study indicate that strains of A. veronii bv veronii produce a heat-labile, pH-stable diarrhoeagenic factor without causing any damage to the intestinal mucosa. PMID- 11014771 TI - Carbachol potentiates cholera toxin-induced secretion in a colonic epithelial cell line (HT29-19A) and rat ileal mucosa in vitro. AB - Recent studies show that enteric nerves are involved in the action of cholera toxin, both in vivo and in vitro. The aim of this study was to investigate in vitro the influence of carbachol, a cholinergic agonist, on the action of cholera toxin. Cultured HT29-19A cell lines and rat ileal mucosa were used in an Ussing chamber for the measurement of short-circuit current induced by cholera toxin. Cyclic AMP was measured from HT29-19A cell lines by standard radio-immunoassay. Pre-treatment of the HT29-19A cell lines with carbachol potentiated cholera toxin induced secretory response, and enhanced accumulation of cAMP. Carbachol also potentiated the cholera toxin-secretory response in the rat ileal mucosa, but only following pretreatment with the prostaglandin synthesis inhibitor, indomethacin. There was synergistic interaction between cholera toxin and cholinergic neurotransmitter carbachol on the intestinal epithelium. Cholinergic agonists may play a role in regulating the secretory response to the toxin. Such interaction is masked in the intact tissues in vitro due to the release of prostaglandins during isolation. PMID- 11014772 TI - Gluten-free diet improves iron-deficiency anaemia in patients with coeliac disease. AB - Two cases of newly-diagnosed asymptomatic coeliac disease with 3 years of unexplained severe iron-deficiency anaemia are presented. Oral iron supplementation had no effect on their serum iron levels and, therefore, had no influence on their anaemia. Upper gastrointestinal endoscopy confirmed normal macroscopic findings. Duodenal biopsies revealed subtotal villous atrophy of the mucosa of the small intestine. A strict gluten-free diet led to an increase in serum iron, resolution of anaemia, and restitution of normal mucosal morphology. Thus, severe iron-deficiency anaemia associated with asymptomatic coeliac disease is responsible to gluten-free diet. PMID- 11014773 TI - Antibiotic resistance conferred by conjugative plasmid in Escherichia coli isolated from community ponds of Kathmandu Valley. PMID- 11014774 TI - Hospitals continue to find robots helpful in foodservice delivery. PMID- 11014775 TI - Espresso kiosks can be profitable addition to hospital foodservice. PMID- 11014776 TI - Atkins diet helps combat diabetes. PMID- 11014777 TI - Physicians drive cost reductions in top orthopaedic unit. PMID- 11014778 TI - Manage patients' pain for better quality, lower costs. PMID- 11014779 TI - System's drug guide reveals costs of branded, generic drugs. PMID- 11014780 TI - Adopt long-stay action board to examine discharge barriers, cut unnecessary hospital days. PMID- 11014781 TI - Greater physician productivity tied to more RNs, support staff. PMID- 11014782 TI - Committing to consistency cuts ventilator days, ICU costs. PMID- 11014783 TI - X-ray quality improvement affects safety, cycle time, patient satisfaction. PMID- 11014785 TI - System revamps ambulatory care to improve patients' experience. PMID- 11014784 TI - Dysfunctional uterine bleeding: new treatment strategies promise lower costs, higher quality. PMID- 11014786 TI - Looking at charges by cost center helps build care plans. PMID- 11014787 TI - Willingness to pay for mammography: item development and testing among five ethnic groups. AB - The goal of this study was to develop a willingness to pay (WTP) question for mammography that is appropriate for low income, ethnically-diverse women. Through qualitative research with 50 low income women of five ethnic groups we developed both a WTP question and a willingness to travel question (WTT). After being refined through interviews with 41 women, these questions were pilot tested on a random sample of 52 low income, ethnically-diverse women in the San Francisco area. Results show that the concepts underlying WTP and WTT were culturally appropriate to the five ethnicities in this study. Analyses generally confirm the validity of the WTP and WTT questions. As expected, WTP was associated with household income, perceived risk of cancer, and knowledge that one needs a mammogram even after a clinical breast examination. Despite the small samples, WTP varied among the ethnic groups. Additionally, WTT was moderately correlated with the natural log of WTP (r = 0.58, P < 0.001). These questions are now in use in a larger clinical trial and future analyses will explore willingness to pay and willingness to travel within and across the ethnic groups. PMID- 11014788 TI - Quick action in ER improves stroke care for all patients. PMID- 11014789 TI - Hoping for improved outcomes, Texas hospital aligns staffing for developmentally supportive care. AB - Take better care of fragile babies with a staff structure that encourages bonding. Texas Children's Hospital in Houston has been making big efforts to provide developmentally supportive care for babies since 1996. Still, neither staff nor physicians thought nurses knew their tiny charges well enough. Texas Children's hopes a new staffing structure will change all that. Discover the details. PMID- 11014791 TI - New website pools wide variety of infection control information. PMID- 11014790 TI - Losing time finding stretchers? Indoor 'GPS' helps hospitals locate equipment, patients. AB - Find missing equipment fast with the indoor equivalent of the global positioning system. Billerica, MA-based PinPoint Co. is marketing a system to locate equipment, such as stretchers and wheelchairs, based on radio-frequency signals emitted from tags on the equipment. A New York hospital that has used the system says it could have a quick payback time. PMID- 11014792 TI - Infection rates drop at hospitals participating in reporting system. AB - Data Benchmarks: The rates of nosocomial and surgical-wound infections have plummeted at hospitals participating in the National Nosocomial Infections Surveillance, a voluntary program of the federal Centers for Disease Control and Prevention. Compare your hospital's performance with the top benchmarks in a new report and learn what organizers say are the essential components of the program's success. PMID- 11014793 TI - System uses pharmacists to take on medication errors. PMID- 11014794 TI - NY system lowers incidence of pressure ulcers with new tool for assessing patients' risk. AB - Preventing pressure ulcers is simple--pay close attention to patients. That means real close. North Shore-Long Island-Jewish Health System shares how it began daily assessments of patients' risks for developing pressure injuries and reduced the number of cases 29% and 80% in the two worst severity levels. PMID- 11014795 TI - Secrets to process improvement revealed: prioritize and finance. AB - To build a better process improvement program, review the processes behind it. There's truth in that tongue-twister: Good ideas for clinical reform often languish if hospitals don't have a structure to support them. Park Nicollet Clinic and Methodist Hospital, an award-winning system in Minneapolis, MN, shares its tips for advancing clinical improvement initiatives. PMID- 11014796 TI - Intense training, monitoring helps carry clinical protocols from trials to actual practice. AB - Can't get a clinical protocol to work in the real world? The problem might lie in training. Intense training and steady monitoring are the keys to getting good results from protocols in actual clinical practice, according to the Center for Behavioral Health in Bloomington, IN. Examining the performance of its panic disorder program, the center found it equaled the results of clinical research trials. PMID- 11014797 TI - Top 100 in cardiac care perform 4.7% to 31% better than others. AB - Data Benchmarks: Measure your hospital against the best in cardiovascular care. A new study from HCIA-Sachs LLC shows that the top-performing cardiovascular programs are ahead of their peers by 4.7% to 31% in eight measures of clinical quality and efficiency. PMID- 11014798 TI - Andre Gratia: a forerunner in microbial and viral genetics. PMID- 11014799 TI - Long-term experimental evolution in Escherichia coli. IX. Characterization of insertion sequence-mediated mutations and rearrangements. AB - As part of a long-term evolution experiment, two populations of Escherichia coli B adapted to a glucose minimal medium for 10,000 generations. In both populations, multiple IS-associated mutations arose that then went to fixation. We identify the affected genetic loci and characterize the molecular events that produced nine of these mutations. All nine were IS-mediated events, including simple insertions as well as recombination between homologous elements that generated inversions and deletions. Sequencing DNA adjacent to the insertions indicates that the affected genes are involved in central metabolism (knockouts of pykF and nadR), cell wall synthesis (adjacent to the promoter of pbpA-rodA), and ill-defined functions (knockouts of hokB-sokB and yfcU). These genes are candidates for manipulation and competition experiments to determine whether the mutations were beneficial or merely hitchhiked to fixation. PMID- 11014800 TI - Saccharomyces cerevisiae BUB2 prevents mitotic exit in response to both spindle and kinetochore damage. AB - The spindle assembly checkpoint-mediated mitotic arrest depends on proteins that signal the presence of one or more unattached kinetochores and prevents the onset of anaphase in the presence of kinetochore or spindle damage. In the presence of either damage, bub2 cells initiate a preanaphase delay but do not maintain it. Inappropriate sister chromatid separation in nocodazole-treated bub2 cells is prevented when mitotic exit is blocked using a conditional tem1(c) mutant, indicating that the preanaphase failure in bub2 cells is a consequence of events downstream of TEM1 in the mitotic exit pathway. Using a conditional bub2(tsd) mutant, we demonstrate that the continuous presence of Bub2 protein is required for maintaining spindle damage-induced arrest. BUB2 is not required to maintain a DNA damage checkpoint arrest, revealing a specificity for spindle assembly checkpoint function. In a yeast two-hybrid assay and in vitro, Bub2 protein interacts with the septin protein Cdc3, which is essential for cytokinesis. These data support the view that the spindle assembly checkpoint encompasses regulation of distinct mitotic steps, including a MAD2-directed block to anaphase initiation and a BUB2-directed block to TEM1-dependent exit. PMID- 11014801 TI - Mutation of the ATP-binding pocket of SSA1 indicates that a functional interaction between Ssa1p and Ydj1p is required for post-translational translocation into the yeast endoplasmic reticulum. AB - The translocation of proteins across the yeast ER membrane requires ATP hydrolysis and the action of DnaK (hsp70) and DnaJ homologues. In Saccharomyces cerevisiae the cytosolic hsp70s that promote post-translational translocation are the products of the Ssa gene family. Ssa1p maintains secretory precursors in a translocation-competent state and interacts with Ydj1p, a DnaJ homologue. Although it has been proposed that Ydj1p stimulates the ATPase activity of Ssa1p to release preproteins and engineer translocation, support for this model is incomplete. To this end, mutations in the ATP-binding pocket of SSA1 were constructed and examined both in vivo and in vitro. Expression of the mutant Ssa1p's slows wild-type cell growth, is insufficient to support life in the absence of functional Ssa1p, and results in a dominant effect on post translational translocation. The ATPase activity of the purified mutant proteins was not enhanced by Ydj1p and the mutant proteins could not bind an unfolded polypeptide substrate. Our data suggest that a productive interaction between Ssa1p and Ydj1p is required to promote protein translocation. PMID- 11014802 TI - Glucose monitoring in fission yeast via the Gpa2 galpha, the git5 Gbeta and the git3 putative glucose receptor. AB - The fission yeast Schizosaccharomyces pombe responds to environmental glucose by activating adenylate cyclase. The resulting cAMP signal activates protein kinase A (PKA). PKA inhibits glucose starvation-induced processes, such as conjugation and meiosis, and the transcription of the fbp1 gene that encodes the gluconeogenic enzyme fructose-1,6-bisphosphatase. We previously identified a collection of git genes required for glucose repression of fbp1 transcription, including pka1/git6, encoding the PKA catalytic subunit, git2/cyr1, encoding adenylate cyclase, and six "upstream" genes required for adenylate cyclase activation. The git8 gene, identical to gpa2, encodes the alpha subunit of a heterotrimeric guanine-nucleotide binding protein (Galpha) while git5 encodes a Gbeta subunit. Multicopy suppression studies with gpa2(+) previously indicated that S. pombe adenylate cyclase activation may resemble that of the mammalian type II enzyme with sequential activation by Galpha followed by Gbetagamma. We show here that an activated allele of gpa2 (gpa2(R176H), carrying a mutation in the coding region for the GTPase domain) fully suppresses mutations in git3 and git5, leading to a refinement in our model. We describe the cloning of git3 and show that it encodes a putative seven-transmembrane G protein-coupled receptor. A git3 deletion confers the same phenotypes as deletions of other components of the PKA pathway, including a germination delay, constitutive fbp1 transcription, and starvation-independent conjugation. Since the git3 deletion is fully suppressed by the gpa2(R176H) allele with respect to fbp1 transcription, git3 appears to encode a G protein-coupled glucose receptor responsible for adenylate cyclase activation in S. pombe. PMID- 11014804 TI - Synthetic lethal interactions suggest a role for the Saccharomyces cerevisiae Rtf1 protein in transcription elongation. AB - Strong evidence indicates that transcription elongation by RNA polymerase II (pol II) is a highly regulated process. Here we present genetic results that indicate a role for the Saccharomyces cerevisiae Rtf1 protein in transcription elongation. A screen for synthetic lethal mutations was carried out with an rtf1 deletion mutation to identify factors that interact with Rtf1 or regulate the same process as Rtf1. The screen uncovered mutations in SRB5, CTK1, FCP1, and POB3. These genes encode an Srb/mediator component, a CTD kinase, a CTD phosphatase, and a protein involved in the regulation of transcription by chromatin structure, respectively. All of these gene products have been directly or indirectly implicated in transcription elongation, indicating that Rtf1 may also regulate this process. In support of this view, we show that RTF1 functionally interacts with genes that encode known elongation factors, including SPT4, SPT5, SPT16, and PPR2. We also show that a deletion of RTF1 causes sensitivity to 6-azauracil and mycophenolic acid, phenotypes correlated with a transcription elongation defect. Collectively, our results suggest that Rtf1 may function as a novel transcription elongation factor in yeast. PMID- 11014803 TI - Suppressors of mdm20 in yeast identify new alleles of ACT1 and TPM1 predicted to enhance actin-tropomyosin interactions. AB - The actin cytoskeleton is required for many aspects of cell division in yeast, including mitochondrial partitioning into growing buds (mitochondrial inheritance). Yeast cells lacking MDM20 function display defects in both mitochondrial inheritance and actin organization, specifically, a lack of visible actin cables and enhanced sensitivity to Latrunculin A. mdm20 mutants also exhibit a temperature-sensitive growth phenotype, which we exploited to isolate second-site suppressor mutations. Nine dominant suppressors selected in an mdm20/mdm20 background rescue temperature-sensitive growth defects and mitochondrial inheritance defects and partially restore actin cables in haploid and diploid mdm20 strains. The suppressor mutations define new alleles of ACT1 and TPM1, which encode actin and the major form of tropomyosin in yeast, respectively. The ACT1 mutations cluster in a region of the actin protein predicted to contact tropomyosin, suggesting that they stabilize actin cables by enhancing actin-tropomyosin interactions. The characteristics of the mutant ACT1 and TPM1 alleles and their potential effects on protein structure and binding are discussed. PMID- 11014805 TI - Involvement of very short DNA tandem repeats and the influence of the RAD52 gene on the occurrence of deletions in Saccharomyces cerevisiae. AB - Chromosomal rearrangements, such as deletions, duplications, or Ty transposition, are rare events. We devised a method to select for such events as Ura(+) revertants of a particular ura2 mutant. Among 133 Ura(+) revertants, 14 were identified as the result of a deletion in URA2. Of seven classes of deletions, six had very short regions of identity at their junctions (from 7 to 13 bp long). This strongly suggests a nonhomologous recombination mechanism for the formation of these deletions. The total Ura(+) reversion rate was increased 4.2-fold in a rad52Delta strain compared to the wild type, and the deletion rate was significantly increased. All the deletions selected in the rad52Delta context had microhomologies at their junctions. We propose two mechanisms to explain the occurrence of these deletions and discuss the role of microhomology stretches in the formation of fusion proteins. PMID- 11014806 TI - A role for cytosolic hsp70 in yeast [PSI(+)] prion propagation and [PSI(+)] as a cellular stress. AB - [PSI(+)] is a prion (infectious protein) of Sup35p, a subunit of the Saccharomyces cerevisiae translation termination factor. We isolated a dominant allele, SSA1-21, of a gene encoding an Hsp70 chaperone that impairs [PSI(+)] mitotic stability and weakens allosuppression caused by [PSI(+)]. While [PSI(+)] stability is normal in strains lacking SSA1, SSA2, or both, SSA1-21 strains with a deletion of SSA2 cannot propagate [PSI(+)]. SSA1-21 [PSI(+)] strains are hypersensitive to curing of [PSI(+)] by guanidine-hydrochloride and partially cured of [PSI(+)] by rapid induction of the heat-shock response but not by growth at 37 degrees. The number of inheritable [PSI(+)] particles is significantly reduced in SSA1-21 cells. SSA1-21 effects on [PSI(+)] appear to be independent of Hsp104, another stress-inducible protein chaperone known to be involved in [PSI(+)] propagation. We propose that cytosolic Hsp70 is important for the formation of Sup35p polymers characteristic of [PSI(+)] from preexisting material and that Ssa1-21p both lacks and interferes with this activity. We further demonstrate that the negative effect of heat stress on [PSI(+)] phenotype directly correlates with solubility of Sup35p and find that in wild-type strains the presence of [PSI(+)] causes a stress that elevates basal expression of Hsp104 and SSA1. PMID- 11014807 TI - Sequence composition and context effects on the generation and repair of frameshift intermediates in mononucleotide runs in Saccharomyces cerevisiae. AB - DNA polymerase slippage occurs frequently in tracts of a tandemly repeated nucleotide, and such slippage events can be genetically detected as frameshift mutations. In long mononucleotide runs, most frameshift intermediates are repaired by the postreplicative mismatch repair (MMR) machinery, rather than by the exonucleolytic proofreading activity of DNA polymerase. Although mononucleotide runs are hotspots for polymerase slippage events, it is not known whether the composition of a run and the surrounding context affect the frequency of slippage or the efficiency of MMR. To address these issues, 10-nucleotide (10N) runs were inserted into the yeast LYS2 gene to create +1 frameshift alleles. Slippage events within these runs were detected as Lys(+) revertants. 10G or 10C runs were found to be more unstable than 10A or 10T runs, but neither the frequency of polymerase slippage nor the overall efficiency of MMR was greatly influenced by sequence context. Although complete elimination of MMR activity (msh2 mutants) affected all runs similarly, analyses of reversion rates in msh3 and msh6 mutants revealed distinct specificities of the yeast Msh2p-Msh3p and Msh2p-Msh6p mismatch binding complexes in the repair of frameshift intermediates in different sequence contexts. PMID- 11014808 TI - Suppression of the profilin-deficient phenotype by the RHO2 signaling pathway in Saccharomyces cerevisiae. AB - Profilin plays an important role in actin organization in all eukaryotic cells through mechanisms that are still poorly understood. We had previously shown that Mid2p, a transmembrane protein and a potential cell wall sensor, is an effective multicopy suppressor of the profilin-deficient phenotype in Saccharomyces cerevisiae. To better understand the role of Mid2p in the organization of the actin cytoskeleton, we isolated five additional multicopy suppressors of pfy1Delta cells that are Rom1p, Rom2p, Rho2p, Smy1p, and the previously uncharacterized protein Syp1p. The problems of caffeine and NaCl sensitivity, growth defects at 30 degrees and 37 degrees, the accumulation of intracellular vesicular structures, and a random budding pattern in pfy1Delta cells are corrected by all the suppressors tested. This is accompanied by a partial repolarization of the cortical actin patches without the formation of visible actin cables. The overexpression of Mid2p, Rom2p, and Syp1p, but not the overexpression of Rho2p and Smy1p, results in an abnormally thick cell wall in wild-type and pfy1Delta cells. Since none of the suppressors, except Rho2p, can correct the phenotype of the pfy1-111/rho2Delta strain, we propose a model in which the suppressors act through the Rho2p signaling pathway to repolarize cortical actin patches. PMID- 11014809 TI - Spore-killing meiotic drive factors in a natural population of the fungus Podospora anserina. AB - In fungi, meiotic drive is observed as spore killing. In the secondarily homothallic ascomycete Podospora anserina it is characterized by the abortion of two of the four spores in the ascus. We have identified seven different types of meiotic drive elements (Spore killers). Among 99 isolates from nature, six of these meiotic drive elements occurred in a local population. Spore killers comprise 23% of the natural population of P. anserina in Wageningen, The Netherlands, sampled from 1991 to 1997. One Spore-killer type was also found in a French strain dating from 1937. All other isolates found so far are sensitive to spore killing. All seven Spore killer types differ in the percentage of asci that show killing and in their mutual interactions. Interactions among Spore killer types showed either mutual resistance or dominant epistasis. Most killer elements could be assigned to linkage group III but are not tightly linked to the centromere. PMID- 11014810 TI - Respiratory chain complex I is essential for sexual development in neurospora and binding of iron sulfur clusters are required for enzyme assembly. AB - We have cloned and disrupted in vivo, by repeat-induced point mutations, the nuclear gene coding for an iron sulfur subunit of complex I from Neurospora crassa, homologue of the mammalian TYKY protein. Analysis of the obtained mutant nuo21.3c revealed that complex I fails to assemble. The peripheral arm of the enzyme is disrupted while its membrane arm accumulates. Furthermore, mutated 21.3c-kD proteins, in which selected cysteine residues were substituted with alanines or serines, were expressed in mutant nuo21. 3c. The phenotypes of these strains regarding the formation of complex I are similar to that of the original mutant, indicating that binding of iron sulfur centers to protein subunits is a prerequisite for complex I assembly. Homozygous crosses of nuo21.3c strain, and of other complex I mutants, are unable to complete sexual development. The crosses are blocked at an early developmental stage, before fusion of the nuclei of opposite mating types. This phenotype can be rescued only by transformation with the intact gene. Our results suggest that this might be due to the compromised capacity of complex I-defective strains in energy production. PMID- 11014811 TI - Caenorhabditis elegans msh-5 is required for both normal and radiation-induced meiotic crossing over but not for completion of meiosis. AB - Crossing over and chiasma formation during Caenorhabditis elegans meiosis require msh-5, which encodes a conserved germline-specific MutS family member. msh-5 mutant oocytes lack chiasmata between homologous chromosomes, and crossover frequencies are severely reduced in both oocyte and spermatocyte meiosis. Artificially induced DNA breaks do not bypass the requirement for msh-5, suggesting that msh-5 functions after the initiation step of meiotic recombination. msh-5 mutants are apparently competent to repair breaks induced during meiosis, but accomplish repair in a way that does not lead to crossovers between homologs. These results combine with data from budding yeast to establish a conserved role for Msh5 proteins in promoting the crossover outcome of meiotic recombination events. Apart from the crossover deficit, progression through meiotic prophase is largely unperturbed in msh-5 mutants. Homologous chromosomes are fully aligned at the pachytene stage, and germ cells survive to complete meiosis and gametogenesis with high efficiency. Our demonstration that artificially induced breaks generate crossovers and chiasmata using the normal meiotic recombination machinery suggests (1) that association of breaks with a preinitiation complex is not a prerequisite for entering the meiotic recombination pathway and (2) that the decision for a subset of recombination events to become crossovers is made after the initiation step. PMID- 11014812 TI - A region of the myosin rod important for interaction with paramyosin in Caenorhabditis elegans striated muscle. AB - The precise arrangement of molecules within the thick filament, as well as the mechanisms by which this arrangement is specified, remains unclear. In this article, we have exploited a unique genetic interaction between one isoform of myosin heavy chain (MHC) and paramyosin in Caenorhabditis elegans to probe the molecular interaction between MHC and paramyosin in vivo. Using chimeric myosin constructs, we have defined a 322-residue region of the MHC A rod critical for suppression of the structural and motility defects associated with the unc 15(e73) allele. Chimeric constructs lacking this region of MHC A either fail to suppress, or act as dominant enhancers of, the e73 phenotype. Although the 322 residue region is required for suppression activity, our data suggest that sequences along the length of the rod also play a role in the isoform-specific interaction between MHC A and paramyosin. Our genetic and cell biological analyses of construct behavior suggest that the 322-residue region of MHC A is important for thick filament stability. We present a model in which this region mediates an avid interaction between MHC A and paramyosin in parallel arrangement in formation of the filament arms. PMID- 11014813 TI - A screen for new trithorax group genes identified little imaginal discs, the Drosophila melanogaster homologue of human retinoblastoma binding protein 2. AB - The proteins encoded by two groups of conserved genes, the Polycomb and trithorax groups, have been proposed to maintain, at the level of chromatin structure, the expression pattern of homeotic genes during Drosophila development. To identify new members of the trithorax group, we screened a collection of deficiencies for intergenic noncomplementation with a mutation in ash1, a trithorax group gene. Five of the noncomplementing deletions uncover genes previously classified as members of the Polycomb group. This evidence suggests that there are actually three groups of genes that maintain the expression pattern of homeotic genes during Drosophila development. The products of the third group appear to be required to maintain chromatin in both transcriptionally inactive and active states. Six of the noncomplementing deficiencies uncover previously unidentified trithorax group genes. One of these deficiencies removes 25D2-3 to 26B2-5. Within this region, there are two, allelic, lethal P-insertion mutations that identify one of these new trithorax group genes. The gene has been called little imaginal discs based on the phenotype of mutant larvae. The protein encoded by the little imaginal discs gene is the Drosophila homologue of human retinoblastoma binding protein 2. PMID- 11014814 TI - Isolation and analysis of six timeless alleles that cause short- or long-period circadian rhythms in Drosophila. AB - In genetic screens for Drosophila mutations affecting circadian locomotion rhythms, we have isolated six new alleles of the timeless (tim) gene. Two of these mutations cause short-period rhythms of 21-22 hr in constant darkness, and four result in long-period cycles of 26-28 hr. All alleles are semidominant. Studies of the genetic interactions of some of the tim alleles with period altering period (per) mutations indicate that these interactions are close to multiplicative; a given allele changes the period length of the genetic background by a fixed percentage, rather than by a fixed number of hours. The tim(L1) allele was studied in molecular detail. The long behavioral period of tim(L1) is reflected in a lengthened molecular oscillation of per and tim RNA and protein levels. The lengthened period is partly caused by delayed nuclear translocation of TIM(L1) protein, shown directly by immunocytochemistry and indirectly by an analysis of the phase response curve of tim(L1) flies. PMID- 11014815 TI - Regulation by homeoproteins: a comparison of deformed-responsive elements. AB - Homeotic genes of Drosophila melanogaster encode transcription factors that specify segment identity by activating the appropriate set of target genes required to produce segment-specific characteristics. Advances in understanding target gene selection have been hampered by the lack of genes known to be directly regulated by the HOM-C proteins. Here we present evidence that the gene 1.28 is likely to be a direct target of Deformed in the maxillary segment. We identified a 664-bp Deformed Response Element (1.28 DRE) that directs maxillary specific expression of a reporter gene in transgenic embryos. The 1.28 DRE contains in vitro binding sites for Deformed and DEAF-1. The Deformed binding sites do not have the consensus sequence for cooperative binding with the cofactor Extradenticle, and we do not detect cooperative binding to these sites, though we cannot rule out an independent role for Extradenticle. Removing the four Deformed binding sites renders the 1.28 DRE inactive in vivo, demonstrating that these sites are necessary for activation of this enhancer element, and supporting the proposition that 1.28 is activated by Deformed. We show that the DEAF-1 binding region is not required for enhancer function. Comparisons of the 1.28 DRE with other known Deformed-responsive enhancers indicate that there are multiple ways to construct Deformed Response Elements. PMID- 11014816 TI - Quantitative trait loci that control vector competence for dengue-2 virus in the mosquito Aedes aegypti. AB - Quantitative trait loci (QTL) affecting the ability of the mosquito Aedes aegypti to become infected with dengue-2 virus were mapped in an F(1) intercross. Dengue susceptible A. aegypti aegypti were crossed with dengue refractory A. aegypti formosus. F(2) offspring were analyzed for midgut infection and escape barriers. In P(1) and F(1) parents and in 207 F(2) individuals, regions of 14 cDNA loci were analyzed with single-strand conformation polymorphism analysis to identify and orient linkage groups with respect to chromosomes I-III. Genotypes were also scored at 57 RAPD-SSCP loci, 5 (TAG)(n) microsatellite loci, and 6 sequence tagged RAPD loci. Dengue infection phenotypes were scored in 86 F(2) females. Two QTL for a midgut infection barrier were detected with standard and composite interval mapping on chromosomes II and III that accounted for approximately 30% of the phenotypic variance (sigma(2)(p)) in dengue infection and these accounted for 44 and 56%, respectively, of the overall genetic variance (sigma(2)(g)). QTL of minor effect were detected on chromosomes I and III, but these were not detected with composite interval mapping. Evidence for a QTL for midgut escape barrier was detected with standard interval mapping but not with composite interval mapping on chromosome III. PMID- 11014817 TI - Male-killing Wolbachia in Drosophila: a temperature-sensitive trait with a threshold bacterial density. AB - Inherited microorganisms that disturb the reproduction of their host have been characterized from a number of host taxa. To understand the general principles underlying the genetic and mechanistic basis of interactions, study of different agents in model host species is required. To this end, the nature and genetics of the maternally inherited sex-ratio trait of Drosophila bifasciata were investigated. Successful curing of affected lines with antibiotics demonstrated this trait was associated with the presence of a bacterium, and molecular systematic analysis demonstrated an association between the presence of the trait and infection with an A group Wolbachia. The penetrance and heritability of the trait did not vary with maternal age. Exposure to elevated temperatures did reduce trait penetrance but did not affect heritability. Examination of the effect of temperature on bacterial density in eggs revealed a decrease in bacterial density following exposure of the parent to elevated temperature, consistent with the hypothesis that male killing in D. bifasciata requires a threshold density of Wolbachia within eggs. The male offspring produced following exposure to elevated temperatures were infected with Wolbachia on emergence as adults. Crossing studies demonstrated a weak cytoplasmic incompatibility phenotype exhibited by Wolbachia in these males. The results are discussed with respect to the incidence of male killing within the clade Wolbachia, the general nature of Wolbachia-host interactions, and the prospects for using this association to investigate the mechanism of male killing. PMID- 11014818 TI - The Drosophila mus101 gene, which links DNA repair, replication and condensation of heterochromatin in mitosis, encodes a protein with seven BRCA1 C-terminus domains. AB - The mutagen-sensitive-101 (mus101) gene of Drosophila melanogaster was first identified 25 years ago through mutations conferring larval hypersensitivity to DNA-damaging agents. Other alleles of mus101 causing different phenotypes were later isolated: a female sterile allele results in a defect in a tissue-specific form of DNA synthesis (chorion gene amplification) and lethal alleles cause mitotic chromosome instability that can be observed genetically and cytologically. The latter phenotype presents as a striking failure of mitotic chromosomes of larval neuroblasts to undergo condensation of pericentric heterochromatic regions, as we show for a newly described mutant carrying lethal allele mus101(lcd). To gain further insight into the function of the Mus101 protein we have molecularly cloned the gene using a positional cloning strategy. We report here that mus101 encodes a member of the BRCT (BRCA1 C terminus) domain superfamily of proteins implicated in DNA repair and cell cycle checkpoint control. Mus101, which contains seven BRCT domains distributed throughout its length, is most similar to human TopBP1, a protein identified through its in vitro association with DNA topoisomerase IIbeta. Mus101 also shares sequence similarity with the fission yeast Rad4/Cut5 protein required for repair, replication, and checkpoint control, suggesting that the two proteins may be functional homologs. PMID- 11014819 TI - Semaphorin-1a acts in concert with the cell adhesion molecules fasciclin II and connectin to regulate axon fasciculation in Drosophila. AB - Semaphorins comprise a large family of phylogenetically conserved secreted and transmembrane glycoproteins, many of which have been implicated in repulsive axon guidance events. The transmembrane semaphorin Sema-1a in Drosophila is expressed on motor axons and is required for the generation of neuromuscular connectivity. Sema-1a can function as an axonal repellent and mediates motor axon defasciculation. Here, by manipulating the levels of Sema-1a and the cell adhesion molecules fasciclin II (Fas II) and connectin (Conn) on motor axons, we provide further evidence that Sema-1a mediates axonal defasciculation events by acting as an axonally localized repellent and that correct motor axon guidance results from a balance between attractive and repulsive guidance cues expressed on motor neurons. PMID- 11014820 TI - Identification of genomic regions that interact with a viable allele of the Drosophila protein tyrosine phosphatase corkscrew. AB - Signaling by receptor tyrosine kinases (RTKs) is critical for a multitude of developmental decisions and processes. Among the molecules known to transduce the RTK-generated signal is the nonreceptor protein tyrosine phosphatase Corkscrew (Csw). Previously, Csw has been demonstrated to function throughout the Drosophila life cycle and, among the RTKs tested, Csw is essential in the Torso, Sevenless, EGF, and Breathless/FGF RTK pathways. While the biochemical function of Csw remains to be unambiguously elucidated, current evidence suggests that Csw plays more than one role during transduction of the RTK signal and, further, the molecular mechanism of Csw function differs depending upon the RTK in question. The isolation and characterization of a new, spontaneously arising, viable allele of csw, csw(lf), has allowed us to undertake a genetic approach to identify loci required for Csw function. The rough eye and wing vein gap phenotypes exhibited by adult flies homo- or hemizygous for csw(lf) has provided a sensitized background from which we have screened a collection of second and third chromosome deficiencies to identify 33 intervals that enhance and 21 intervals that suppress these phenotypes. We have identified intervals encoding known positive mediators of RTK signaling, e.g., drk, dos, Egfr, E(Egfr)B56, pnt, Ras1, rolled/MAPK, sina, spen, Src64B, Star, Su(Raf)3C, and vein, as well as known negative mediators of RTK signaling, e.g., aos, ed, net, Src42A, sty, and su(ve). Of particular interest are the 5 lethal enhancing intervals and 14 suppressing intervals for which no candidate genes have been identified. PMID- 11014821 TI - The LAMMER protein kinase encoded by the Doa locus of Drosophila is required in both somatic and germline cells and is expressed as both nuclear and cytoplasmic isoforms throughout development. AB - Activity of the Darkener of apricot (Doa) locus of Drosophila melanogaster is required for development of the embryonic nervous system, segmentation, photoreceptor maintenance, normal transcription, and sexual differentiation. The gene encodes a protein kinase, with homologues throughout eukaryotes known as the LAMMER kinases. We show here that DOA is expressed as at least two different protein isoforms of 105 and 55 kD throughout development, which are primarily localized to the cytoplasm and nucleus, respectively. Doa transcripts and protein are expressed in all cell types both during embryogenesis and in imaginal discs. Although it was recently shown that DOA kinase is essential for normal sexual differentiation, levels of both kinase isoforms are equal between the sexes during early pupal development. The presence of the kinase on the cell membrane and in the nuclei of polytene salivary gland cells, as well as exclusion from the nuclei of specific cells, may be indicative of regulated kinase localization. Mosaic analysis in both the soma and germline demonstrates that Doa function is essential for cell viability. Finally, in contrast to results reported in other systems and despite some phenotypic similarities, genetic data demonstrate that the LAMMER kinases do not participate in the ras-MAP kinase signal transduction pathway. PMID- 11014822 TI - Identification of autosomal regions involved in Drosophila Raf function. AB - Raf is an essential downstream effector of activated p21(Ras) (Ras) in transducing proliferation or differentiation signals. Following binding to Ras, Raf is translocated to the plasma membrane, where it is activated by a yet unidentified "Raf activator." In an attempt to identify the Raf activator or additional molecules involved in the Raf signaling pathway, we conducted a genetic screen to identify genomic regions that are required for the biological function of Drosophila Raf (Draf). We tested a collection of chromosomal deficiencies representing approximately 70% of the autosomal euchromatic genomic regions for their abilities to enhance the lethality associated with a hypomorphic viable allele of Draf, Draf(Su2). Of the 148 autosomal deficiencies tested, 23 behaved as dominant enhancers of Draf(Su2), causing lethality in Draf(Su2) hemizygous males. Four of these deficiencies identified genes known to be involved in the Drosophila Ras/Raf (Ras1/Draf) pathway: Ras1, rolled (rl, encoding a MAPK), 14-3-3epsilon, and bowel (bowl). Two additional deficiencies removed the Drosophila Tec and Src homologs, Tec29A and Src64B. We demonstrate that Src64B interacts genetically with Draf and that an activated form of Src64B, when overexpressed in early embryos, causes ectopic expression of the Torso (Tor) receptor tyrosine kinase-target gene tailless. In addition, we show that a mutation in Tec29A partially suppresses a gain-of-function mutation in tor. These results suggest that Tec29A and Src64B are involved in Tor signaling, raising the possibility that they function to activate Draf. Finally, we discovered a genetic interaction between Draf(Su2) and Df(3L)vin5 that revealed a novel role of Draf in limb development. We find that loss of Draf activity causes limb defects, including pattern duplications, consistent with a role for Draf in regulation of engrailed (en) expression in imaginal discs. PMID- 11014823 TI - Nonrandom segregation of the mouse univalent X chromosome: evidence of spindle mediated meiotic drive. AB - A fundamental principle of Mendelian inheritance is random segregation of alleles to progeny; however, examples of distorted transmission either of specific alleles or of whole chromosomes have been described in a variety of species. In humans and mice, a distortion in chromosome transmission is often associated with a chromosome abnormality. One such example is the fertile XO female mouse. A transmission distortion effect that results in an excess of XX over XO daughters among the progeny of XO females has been recognized for nearly four decades. Utilizing contemporary methodology that combines immunofluorescence, FISH, and three-dimensional confocal microscopy, we have readdressed the meiotic segregation behavior of the single X chromosome in oocytes from XO females produced on two different inbred backgrounds. Our studies demonstrate that segregation of the univalent X chromosome at the first meiotic division is nonrandom, with preferential retention of the X chromosome in the oocyte in approximately 60% of cells. We propose that this deviation from Mendelian expectations is facilitated by a spindle-mediated mechanism. This mechanism, which appears to be a general feature of the female meiotic process, has implications for the frequency of nondisjunction in our species. PMID- 11014824 TI - Evolution of odorant receptors expressed in mammalian testes. AB - About 10% of mammalian odorant receptors are transcribed in testes, and odorant receptor proteins have been detected on mature spermatozoa. Testis-expressed odorant receptors (TORs) are hypothesized to play roles in sperm chemotaxis, but they might also be ordinary nasal odorant receptors (NORs) that are expressed gratuitously in testes. Under the sperm-chemotaxis hypothesis, TORs should be subject to intense sexual selection and therefore should show higher rates of amino acid substitution than NORs, but under the gratuitous-expression hypothesis, TORs are misidentified NORs and therefore should evolve like other NORs. To test these predictions, we estimated synonymous and nonsynonymous divergences of orthologous NOR and TOR coding sequences from rat and mouse. Contrary to both hypotheses, TORs are on average more highly conserved than NORs, especially in certain domains of the OR protein. This pattern suggests that some TORs might perform internal nonolfactory functions in testes; for example, they might participate in the regulation of sperm development. However, the pattern is also consistent with a modified gratuitous-expression model in which NORs with specialized ligand specificities are both more highly conserved than typical NORs and more likely to be expressed in testes. PMID- 11014825 TI - Archaic lineages in the history of modern humans. AB - An important question in the ongoing debate on the origin of Homo sapiens is whether modern human populations issued from a single lineage or whether several, independently evolving lineages contributed to their genetic makeup. We analyzed haplotypes composed of 35 polymorphisms from a segment of the dystrophin gene. We find that the bulk of a worldwide sample of 868 chromosomes represents haplotypes shared by different continental groups. The remaining chromosomes carry haplotypes specific for the continents or for local populations. The haplotypes specific for non-Africans can be derived from the most frequent ones through simple recombination or a mutation. In contrast, chromosomes specific for sub Saharan Africans represent a distinct group, as shown by principal component analysis, maximum likelihood tree, structural comparison, and summary statistics. We propose that African chromosomes descend from at least two lineages that have been evolving separately for a period of time. One of them underwent range expansion colonizing different continents, including Africa, where it mixed with another, local lineage represented today by a large fraction of African-specific haplotypes. Genetic admixture involving archaic lineages appears therefore to have occurred within Africa rather than outside this continent, explaining greater diversity of sub-Saharan populations observed in a variety of genetic systems. PMID- 11014827 TI - Gene flow in a facultative apomictic poacea, the savanna grass Hyparrhenia diplandra. AB - The genetics of the poacea Hyparrhenia diplandra was studied in four natural populations from an ecological station in West Africa, where it makes up 80% of grasses from wet savanna and constitutes a dense continuum of randomly distributed individuals. DNA content and cytogenetical observations suggest it is an allotetraploid. Using two highly variable microsatellites (heterozygosity H = 0.615-0.616), we show that this species is an apomict with rare sexual reproduction events that account for approximately 0.5% of seeds pollinated in the wild. Hexaploid individuals were also produced, corroborating the observation of aberrant genotypes in the wild. The spatial extent of asexual clones in the field was low in comparison with the predominance of apomixis, thus indicating a low dispersal of seeds from their parent. Heterozygosity and departure from Hardy Weinberg predictions were similar in the four populations, revealing a high apparent selfing rate s = 0.599 among sexually produced seeds. This is an overestimate since we could not distinguish true selfing from reciprocal outcrosses between neighboring individuals from the same apomictic clone. Gene flow by pollen could be substantial, possibly explaining the absence of isolation by distance in the studied area. PMID- 11014826 TI - Mechanisms involved in targeted gene replacement in mammalian cells. AB - The "ends-out" or omega (Omega)-form gene replacement vector is used routinely to perform targeted genome modification in a variety of species and has the potential to be an effective vehicle for gene therapy. However, in mammalian cells, the frequency of this reaction is low and the mechanism unknown. Understanding molecular features associated with gene replacement is important and may lead to an increase in the efficiency of the process. In this study, we investigated gene replacement in mammalian cells using a powerful assay system that permits efficient recovery of the product(s) of individual recombination events at the haploid, chromosomal mu-delta locus in a murine hybridoma cell line. The results showed that (i) heteroduplex DNA (hDNA) is formed during mammalian gene replacement; (ii) mismatches in hDNA are usually efficiently repaired before DNA replication and cell division; (iii) the gene replacement reaction occurs with fidelity; (iv) the presence of multiple markers in one homologous flanking arm in the replacement vector did not affect the efficiency of gene replacement; and (v) in comparison to a genomic fragment bearing contiguous homology to the chromosomal target, gene targeting was only slightly inhibited by internal heterology (pSV2neo sequences) in the replacement vector. PMID- 11014828 TI - Comparative fluorescence in situ hybridization mapping of a 431-kb Arabidopsis thaliana bacterial artificial chromosome contig reveals the role of chromosomal duplications in the expansion of the Brassica rapa genome. AB - Comparative genome studies are important contributors to our understanding of genome evolution. Most comparative genome studies in plants have been based on genetic mapping of homologous DNA loci in different genomes. Large-scale comparative physical mapping has been hindered by the lack of efficient and affordable techniques. We report here the adaptation of fluorescence in situ hybridization (FISH) techniques for comparative physical mapping between Arabidopsis thaliana and Brassica rapa. A set of six bacterial artificial chromosomes (BACs) representing a 431-kb contiguous region of chromosome 2 of A. thaliana was mapped on both chromosomes and DNA fibers of B. rapa. This DNA fragment has a single location in the A. thaliana genome, but hybridized to four to six B. rapa chromosomes, indicating multiple duplications in the B. rapa genome. The sizes of the fiber-FISH signals from the same BACs were not longer in B. rapa than those in A. thaliana, suggesting that this genomic region is duplicated but not expanded in the B. rapa genome. The comparative fiber-FISH mapping results support that chromosomal duplications, rather than regional expansion due to accumulation of repetitive sequences in the intergenic regions, played the major role in the evolution of the B. rapa genome. PMID- 11014829 TI - Hierarchical patterns of transgene expression indicate involvement of developmental mechanisms in the regulation of the maize P1-rr promoter. AB - The maize P1-rr gene encodes a Myb-homologous transcription factor that regulates the synthesis of red flavonoid pigments. Maize plants transformed with segments of the P1-rr promoter driving a GUS reporter gene exhibit significant variation in transgene expression, both between independent transformation events and among sibling plants derived from a single event. Interestingly, variability in spatial expression is not random; rather, transgene activity occurs predominantly in five patterns that fit a hierarchy: expression is most common in kernel pericarp, with sequential addition of expression in cob glumes, husk, silk, and tassel. The hierarchical expression pattern of P-rr::GUS transgenes suggests a possible model for developmental regulation of the P1-rr gene. Our results demonstrate that variability in transgene expression, a common occurrence in transgenic plant studies, can be informative if adequately analyzed to uncover underlying patterns of gene expression. PMID- 11014830 TI - Computational and experimental characterization of physically clustered simple sequence repeats in plants. AB - The type and frequency of simple sequence repeats (SSRs) in plant genomes was investigated using the expanding quantity of DNA sequence data deposited in public databases. In Arabidopsis, 306 genomic DNA sequences longer than 10 kb and 36,199 EST sequences were searched for all possible mono- to pentanucleotide repeats. The average frequency of SSRs was one every 6.04 kb in genomic DNA, decreasing to one every 14 kb in ESTs. SSR frequency and type differed between coding, intronic, and intergenic DNA. Similar frequencies were found in other plant species. On the basis of these findings, an approach is proposed and demonstrated for the targeted isolation of single or multiple, physically clustered SSRs linked to any gene that has been mapped using low-copy DNA-based markers. The approach involves sample sequencing a small number of subclones of selected randomly sheared large insert DNA clones (e.g., BACs). It is shown to be both feasible and practicable, given the probability of fortuitously sequencing through an SSR. The approach is demonstrated in barley where sample sequencing 34 subclones of a single BAC selected by hybridization to the Big1 gene revealed three SSRs. These allowed Big1 to be located at the top of barley linkage group 6HS. PMID- 11014831 TI - On the differences between maximum likelihood and regression interval mapping in the analysis of quantitative trait loci. AB - The differences between maximum-likelihood (ML) and regression (REG) interval mapping in the analysis of quantitative trait loci (QTL) are investigated analytically and numerically by simulation. The analytical investigation is based on the comparison of the solution sets of the ML and REG methods in the estimation of QTL parameters. Their differences are found to relate to the similarity between the conditional posterior and conditional probabilities of QTL genotypes and depend on several factors, such as the proportion of variance explained by QTL, relative QTL position in an interval, interval size, difference between the sizes of QTL, epistasis, and linkage between QTL. The differences in mean squared error (MSE) of the estimates, likelihood-ratio test (LRT) statistics in testing parameters, and power of QTL detection between the two methods become larger as (1) the proportion of variance explained by QTL becomes higher, (2) the QTL locations are positioned toward the middle of intervals, (3) the QTL are located in wider marker intervals, (4) epistasis between QTL is stronger, (5) the difference between QTL effects becomes larger, and (6) the positions of QTL get closer in QTL mapping. The REG method is biased in the estimation of the proportion of variance explained by QTL, and it may have a serious problem in detecting closely linked QTL when compared to the ML method. In general, the differences between the two methods may be minor, but can be significant when QTL interact or are closely linked. The ML method tends to be more powerful and to give estimates with smaller MSEs and larger LRT statistics. This implies that ML interval mapping can be more accurate, precise, and powerful than REG interval mapping. The REG method is faster in computation, especially when the number of QTL considered in the model is large. Recognizing the factors affecting the differences between REG and ML interval mapping can help an efficient strategy, using both methods in QTL mapping to be outlined. PMID- 11014833 TI - Consequences of recombination on traditional phylogenetic analysis. AB - We investigate the shape of a phylogenetic tree reconstructed from sequences evolving under the coalescent with recombination. The motivation is that evolutionary inferences are often made from phylogenetic trees reconstructed from population data even though recombination may well occur (mtDNA or viral sequences) or does occur (nuclear sequences). We investigate the size and direction of biases when a single tree is reconstructed ignoring recombination. Standard software (PHYLIP) was used to construct the best phylogenetic tree from sequences simulated under the coalescent with recombination. With recombination present, the length of terminal branches and the total branch length are larger, and the time to the most recent common ancestor smaller, than for a tree reconstructed from sequences evolving with no recombination. The effects are pronounced even for small levels of recombination that may not be immediately detectable in a data set. The phylogenies when recombination is present superficially resemble phylogenies for sequences from an exponentially growing population. However, exponential growth has a different effect on statistics such as Tajima's D. Furthermore, ignoring recombination leads to a large overestimation of the substitution rate heterogeneity and the loss of the molecular clock. These results are discussed in relation to viral and mtDNA data sets. PMID- 11014832 TI - The implications of intergenic polymorphism for major histocompatibility complex evolution. AB - A systematic survey of six intergenic regions flanking the human HLA-B locus in eight haplotypes reveals the regions to be up to 20 times more polymorphic than the reported average degree of human neutral polymorphism. Furthermore, the extent of polymorphism is directly related to the proximity to the HLA-B locus. Apparently linkage to HLA-B locus alleles, which are under balancing selection, maintains the neutral polymorphism of adjacent regions. For these linked polymorphisms to persist, recombination in the 200-kb interval from HLA-B to TNF must occur at a low frequency. The high degree of polymorphism found distal to HLA-B suggests that recombination is uncommon on both sides of the HLA-B locus. The least-squares estimate is 0.15% per megabase with an estimated range from 0.02 to 0.54%. These findings place strong restrictions on possible recombinational mechanisms for the generation of diversity at the HLA-B. PMID- 11014834 TI - The evolution of haploid, diploid and polymorphic haploid-diploid life cycles: the role of meiotic mutation. AB - Here I present a simple population genetic model to investigate the evolution of polymorphic haploid-diploid life cycles. The key feature of the model is the assumption of mutation occurring during meiosis. I show that, in addition to regions favoring haploid or diploid life cycles, there are substantial regions of the parameter space under which polymorphic haploid-diploid life cycles are expected to evolve. PMID- 11014835 TI - Multitrait least squares for quantitative trait loci detection. AB - A multiple-trait QTL mapping method using least squares is described. It is presented as an extension of a single-trait method for use with three-generation, outbred pedigrees. The multiple-trait framework allows formal testing of whether the same QTL affects more than one trait (i.e., a pleiotropic QTL) or whether more than one linked QTL are segregating. Several approaches to the testing procedure are presented and their suitability discussed. The performance of the method is investigated by simulation. As previously found, multitrait analyses increase the power to detect a pleiotropic QTL and the precision of its location estimate. With enough information, discrimination between alternative genetic models is possible. PMID- 11014836 TI - Statistical models for estimating the genetic basis of repeated measures and other function-valued traits. AB - The genetic analysis of characters that are best considered as functions of some independent and continuous variable, such as age, can be a complicated matter, and a simple and efficient procedure is desirable. Three methods are common in the literature: random regression, orthogonal polynomial approximation, and character process models. The goals of this article are (i) to clarify the relationships between these methods; (ii) to develop a general extension of the character process model that relaxes correlation stationarity, its most stringent assumption; and (iii) to compare and contrast the techniques and evaluate their performance across a range of actual and simulated data. We find that the character process model, as described in 1999 by Pletcher and Geyer, is the most successful method of analysis for the range of data examined in this study. It provides a reasonable description of a wide range of different covariance structures, and it results in the best models for actual data. Our analysis suggests genetic variance for Drosophila mortality declines with age, while genetic variance is constant at all ages for reproductive output. For growth in beef cattle, however, genetic variance increases linearly from birth, and genetic correlations are high across all observed ages. PMID- 11014837 TI - Simple and rapid method for the detection of early signs of toxicity in Daphnia magna straus. PMID- 11014838 TI - Endocrine disruption by hexachlorobenzene in Crucian carp (Carassius auratus gibelio). PMID- 11014841 TI - Lead concentrations in Eucalyptus sp. in a small coastal town. PMID- 11014839 TI - Toxicity response of Emerita analoga (Stimpson, 1857) collected from beaches of South Central Chile. PMID- 11014842 TI - Metals in water in the Central Kalimantan, Indonesia. PMID- 11014843 TI - Migration of lead and cadmium from ceramic materials used in food preparation. PMID- 11014840 TI - Mercury bioaccumulation induces oxidative stress and toxicity to submerged macrophyte Potamogeton crispus L. PMID- 11014844 TI - Metals and selenium in Sand Martin's plumage. PMID- 11014846 TI - Bioconcentration of endosulfan and monocrotophos by Labeo rohita and Channa punctata. PMID- 11014847 TI - Toxicity and bioconcentration of chlorpyrifos in aquatic organisms: Artemia parthenogenetica(Crustacea), Gambusia affinis, and Aphanius iberus (Pisces). PMID- 11014845 TI - Determination of the bioconcentration of phosphamidon and profenofos in zebrafish (Brachydanio rerio). PMID- 11014848 TI - Measurement of benzo(a)pyrene in sea water and in mussels in the Seto Inland Sea, Japan. PMID- 11014853 TI - Comparison of different treatments for alachlor removal from water. PMID- 11014852 TI - Identification of hydrocarbons and oxygen compounds in sediments from Niigata, Japan. PMID- 11014849 TI - Cadmium uptake by Corbicula fluminea and Dreissena polymorpha: effects of pH and temperature. PMID- 11014850 TI - Time-related increase of hydrocarbons in barnacles in the north-western waters of the Arabian gulf. PMID- 11014854 TI - Management of fly ash landfills with Cassia surattensis Burm: a case study. PMID- 11014856 TI - Fluctuations and fractal noise in biological membranes. AB - Our understanding of cell structure and function derives from applications of a variety of physical and life science disciplines, methods and models to an important physiological process, namely, the exchange and transport of ions and molecules across biological membranes. We know that ion transport through membranes arises from a diversity of interrelated and interactive physical and chemical phenomena over a wide range of spatial and temporal scales. Among these phenomena common to all cellular structure and function include metabolism, kinetics of molecules, chemically mediated alteration of cell membrane electrical potential, membrane ion conductance, electrical signal propagation, and modulation by chemo- and mechanoreceptive mechanisms. This review focuses on the unique information contained in fluctuations in electrical properties associated with cell membrane ion transport. PMID- 11014855 TI - Fluoride removal from water by Hydrilla verticillata (l.f.) royle and its toxic effects. PMID- 11014851 TI - Surveys of volatile organic compounds in soil and groundwater at industrial sites in Taiwan. PMID- 11014857 TI - Reactivity of cysteines in the transmembrane region of the Na, K-ATPase alpha subunit probed with Hg(2+). AB - To gain insight into the structure and conformational coupling in the Na,K ATPase, this study characterized the reaction of the alpha1 subunit transmembrane cysteines with a small probe. Intact HeLa cells expressing heterologous Na,K ATPase were treated with (microm) HgCl(2) after placing the enzyme predominantly in either of two conformations, phosphorylated E2P.Na/E2P or dephosphorylated ATP.E1. K/ATP.E1. Under both conditions the treatment led to enzyme inactivation following a double exponential kinetic as determined by ouabain-sensitive K(+) uptake measurements. However, the rate constant of the slow reacting component was ten times larger when the protein was probed in a medium that would favor enzyme phosphorylation. Enzymes carrying mutations of cysteines located in the alpha1 subunit transmembrane region were used to identify the reacting-SH groups. Replacement Cys104Ser reduced enzyme inactivation by removing the slow reacting component under both treatment conditions. Replacement of Cys964 reduced the inactivation rate constant of the fast reacting component (79%) and removed the slow reacting component when the dephosphorylated enzyme was treated with Hg(2+). Moreover, Cys964Ser substituted enzyme was insensitive to Hg(2+) when treated under phosphorylation conditions. These results indicate that Cys964 is involved in the fast inactivation by Hg(2+). Although the double mutant Cys964, 104Ser was still partially inactivated by treatment under nonphosphorylating conditions, an enzyme devoid of transmembrane cysteines was insensitive to Hg(2+) under all treatment conditions. Thus, this enzyme provides a background where accessibility of engineered transmembrane cysteines can be tested. PMID- 11014861 TI - Mechanism of shrinkage activation of nonselective cation channels in M-1 mouse cortical collecting duct cells. AB - It has previously been shown that osmotic cell shrinkage activates a nonselective cation (NSC) channel in M-1 mouse cortical collecting duct cells [54] and in a variety of other cell types [20]. In the present study we further characterized the shrinkage-activated NSC channel in M-1 cells and its mechanism of activation using whole-cell current recordings. Osmotic cell shrinkage induced by addition of 100 mm sucrose to the bath solution caused a 20-fold increase in whole-cell inward currents from -10.8 +/- 1.5 pA to -211 +/- 10.2 pA (n = 103). A similar response was observed when cell shrinkage was elicited using a hypo-osmotic pipette solution. This indicates that cell shrinkage and not extracellular osmolarity per se is the signal for current activation. Cation substitution experiments revealed that the activated channels discriminate poorly between monovalent cations with a selectivity sequence NH(4) (1.2) > or = Na(+) (1) approximately K(+) (0.9) approximately Li(+) (0.9). In contrast there was no measurable permeability for Ca(2+) or Ba(2+) and the cation-to-anion permeability ratio was about 14. The DPC-derivatives flufenamic acid, 4-methyl-DPC and DCDPC were the most effective blockers followed by LOE 908, while amiloride and bumetanide were ineffective. The putative channel activator maitotoxin had no effect. Current activation was dependent upon the presence of intracellular ATP and Mg(2+) and was inhibited by staurosporine (1 microm) and calphostin C (1 microm). Moreover, cytochalasin D (10 microm) and taxol (2 microm) reduced the current response to cell shrinkage. These findings suggest that the activation mechanism of the shrinkage-activated NSC channel involves protein kinase mediated phosphorylation steps and cytoskeletal elements. PMID- 11014863 TI - The SU glycoprotein 120 from HIV-1 penetrates into lipid monolayers mimicking plasma membranes. AB - Increasing evidence suggests that the HIV envelope binds through its surface (SU) gp120 not only to receptors and coreceptors, but also to other components of the cellular membrane where the glycolipids appear to be good candidates. To assess the ability of HIV-1 SU gp120 to penetrate into phospholipid membranes, we carried out a study of the interactions between a recombinant SU gp120 from HIV 1/HXB2 and artificial lipid monolayers mimicking the composition of the outer leaflet of the lymphocytes and which were spread at the air-water interface. We show that the protein, in its aggregated form, has amphipathic properties and that the insertion of this amphipathic species into lipids is favored by the presence of sphingomyelin. Furthermore, cholesterol enhances the penetration into mixed phosphatidylcholine-sphingomyelin monolayers. Coexistence of different physical states of the lipids and thus of domains appears to play a major role for protein penetration independently of the presence of receptors and coreceptors. PMID- 11014858 TI - Vacuolar chloride transport in Mesembryanthemum crystallinum L. measured using the fluorescent dye lucigenin. AB - To study vacuolar chloride (Cl(-)) transport in the halophilic plant Mesembryanthemum crystallinum L., Cl(-) uptake into isolated tonoplast vesicles was measured using the Cl(-)-sensitive fluorescent dye lucigenin (N,N'-dimethyl 9,9'-bisacridinium dinitrate). Lucigenin was used at excitation and emission wavelengths of 433 nm and 506 nm, respectively, and showed a high sensitivity towards Cl(-), with a Stern-Volmer constant of 173 m(-1) in standard assay buffer. While lucigenin fluorescence was strongly quenched by all halides, it was only weakly quenched, if at all, by other anions. However, the fluorescence intensity and Cl(-)-sensitivity of lucigenin was shown to be strongly affected by alkaline pH and was dependent on the conjugate base used as the buffering ion. Chloride transport into tonoplast vesicles of M. crystallinum loaded with 10 mm lucigenin showed saturation-type kinetics with an apparent K(m) of 17.2 mm and a V(max) of 4.8 mm min(-1). Vacuolar Cl(-) transport was not affected by sulfate, malate, or nitrate. In the presence of 250 microm p-chloromercuribenzene sulfonate, a known anion-transport inhibitor, vacuolar Cl(-) transport was actually significantly increased by 24%. To determine absolute fluxes of Cl(-) using this method, the average surface to volume ratio of the tonoplast vesicles was measured by electron microscopy to be 1.13 x 10(7) m(-1). After correcting for a 4.4-fold lower apparent Stern-Volmer constant for intravesicular lucigenin, a maximum rate of Cl(-) transport of 31 nmol m(-2) sec(-1) was calculated, in good agreement with values obtained for the plant vacuolar membrane using other techniques. PMID- 11014859 TI - CFTR regulation of intracellular calcium in normal and cystic fibrosis human airway epithelia. AB - In cystic fibrosis airway epithelia, mutation of the CFTR protein causes a reduced response of Cl(-) secretion to secretagogues acting via cAMP. Using a Ca(2+) imaging system, the hypothesis that CFTR activation may permit ATP release and regulate [Ca(2+)](i) via a receptor-mediated mechanism, is tested in this study. Application of external nucleotides produced a significant increase in [Ca(2+)](i) in normal (16HBE14o(-) cell line and primary lung culture) and in cystic fibrosis (CFTE29o(-) cell line) human airway epithelia. The potency order of nucleotides on [Ca(2+)](i) variation was UTP >> ATP > UDP > ADP > AMP > adenosine in both cell types. The nucleotide [Ca(2+)](i) response could be mimicked by activation of CFTR with forskolin (20 microm) in a temperature dependent manner. In 16HBE14o(-) cells, the forskolin-induced [Ca(2+)](i) response increased with increasing temperature. In CFTE29o(-) cells, forskolin had no effect on [Ca(2+)](i) at body temperature-forskolin-induced [Ca(2+)](i) response in CF cells could only be observed at low experimental temperature (14 degrees C) or when cells were cultured at 26 degrees C instead of 37 degrees C. Pretreatment with CFTR channel blockers glibenclamide (100 microm) and DPC (100 microm), with hexokinase (0.5 U/mg), and with the purinoceptor antagonist suramin (100 microm), inhibited the forskolin [Ca(2+)](i) response. Together, these results demonstrate that once activated, CFTR regulates [Ca(2+)](i) by mediating nucleotide release and activating cell surface purinoceptors in normal and CF human airway epithelia. PMID- 11014862 TI - Inositol 1,4,5-trisphosphate but not ryanodine-receptor agonists induces calcium release from rat liver Golgi apparatus membrane vesicles. AB - We investigated the direct effect of inositol 1,4,5-trisphosphate (IP(3)) and ryanodine receptor agonists on Ca(2+) release from vesicles of a rat liver Golgi apparatus (GA) enriched fraction, which were actively loaded with (45)Ca(2+). Results in GA were compared with those obtained in a rat liver endoplasmic reticulum (ER) enriched fraction. The addition of IP(3) at concentrations ranging from 100 nm to 100 microm, in the presence of thapsigargin, a specific inhibitor of sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPases, promoted a rapid decrease in the Ca(2+) content of GA vesicles. The amount of Ca(2+) released from the vesicles was a function of IP(3) concentration, reaching about 60% in both GA and ER fractions at 100 microm IP(3). Calcium release was inhibited by heparin, an antagonist of IP(3) receptors. Calcium exhibited a bell-shaped effect on IP(3) dependent Ca(2+) released from GA vesicles: it activated Ca(2+) release at concentrations up to 1 microm, and inhibited it at higher concentrations. In contrast to that found in the endoplasmic reticulum fraction, none of the ryanodine receptor agonists tested (cyclic ADP-ribose, caffeine and ryanodine) significantly induced Ca(2+) release from GA fraction vesicles in the presence of thapsigargin. Our results indicate the presence of an IP(3)-sensitive Ca(2+) release mechanism in the Golgi apparatus membrane analogous to that of the ER. However, a Ca(2+) release mechanism sensitive to ryanodine receptor agonists like that of ER is not evident in the GA membrane. PMID- 11014860 TI - Cl(-) channels in basolateral TAL membranes XV. Molecular heterogeneity between cortical and medullary channels. AB - We have isolated two new and highly homologous cDNAs, mmClC-Ka from mouse outer medulla and mcClC-Ka from mouse cortex. In both cases, mRNA was obtained from the indicated region and subjected to RT-PCR using primers from the nucleotide sequence of rbClC-Ka, which encodes basolateral Cl(-) channels (termed rbClC-Ka) in rabbit MTAL. The predicted protein products of mmClC-Ka and mcClC-Ka, mmClC-Ka and mcClC-Ka, respectively, were 85% homologous and had predicted molecular weights of 75 kDa. The predicted protein sequences for mmClC-Ka and rbClC-Ka had three cytosolic sites-threonine 185, threonine 187 and serine 270-which were absent in mcClC-Ka. These three moieties represent potential sites for phosphorylation of mmClC-Ka and rbClC-Ka, but not of mcClC-Ka, and may account for the failure of (ATP + PKA) to increase the open time probability P(o) in basolateral CTAL Cl(-) channels. We prepared antisense oligonucleotides specific for nonhomologous regions of these two cDNAs, mmAntisense for mmClC-Ka and mcAntisense for mcClC-Ka. Using anti-rbClC-Ka, a polyclonal antibody to rbClC-Ka, we found that, when transfected into cultured mouse MTAL and CTAL cells, mmAntisense suppressed the appearance of the 75 kDa band by 50% in vesicles from MTAL but not CTAL cells, while transfection of MTAL and CTAL cells with mcAntisense suppressed appearance of the 75 kDa band in vesicles from CTAL but not MTAL cells. mmAntisense transfection also prolonged the half-time (T(1/2), sec) for (36)Cl(-) efflux in cultured MTAL cells from 82.4 +/- 6.8 sec (sem) to 187.8 +/- 9.5 sec (n = 5; P = 0.0001) while mcAntisense transfection had no such effect. Conversely, in cultured CTAL cells, mcAntisense transfection prolonged the T(1/2) for (36)Cl(-) efflux from 80.9 +/- 6.3 sec to 191.8 +/- 6.5 sec (n = 5; P = 0.00005), while mmAntisense had no such effect. We conclude that mmClC-Ka and mcClC-Ka may encode the basolateral Cl(-) channels mediating net Cl(-) absorption in mouse MTAL and CTAL, respectively. PMID- 11014864 TI - Properties of hexadecaprenyl monophosphate/dioleoylphosphatidylcholine vesicular lipid bilayers. AB - In our study we investigated hemispherical phospholipid bilayer membranes and phospholipid vesicles made from hexadecaprenyl monophosphate (C(80)-P), dioleoylphosphatidylocholine (DOPC) and their mixtures by voltammetric and transmission electron microscopy (TEM) techniques. The current-voltage characteristics, the membrane conductance-temperature relationships and the membrane breakdown voltage have been measured for different mixtures of C(80) P/DOPC. The membrane hydrophobic thickness and the activation energy of ion migration across the membrane have been determined. Hexadecaprenyl monophosphate decreased in comparison with DOPC bilayers, the membrane conductance, increased the activation energy and the membrane breakdown voltage for the various value of C(80)-P/DOPC mole ratio, respectively. The TEM micrographs of C(80)-P, DOPC and C(80)-P/DOPC lipid vesicles showed several characteristic structures, which have been described. The data indicate that hexadecaprenyl monophosphate modulates the surface curvature of the membranes by the formation of aggregates in liquid crystalline phospholipid membranes. We suggest that the dynamics and conformation of hexadecaprenyl monophosphate in membranes depend on the transmembrane electrical potential. The electron micrographs indicate that polyprenyl monophosphates with single isoprenyl chains form lipid vesicular bilayers. The thickness of the bilayer, evaluated from the micrographs, was 11 +/- 1 nm. This property creates possibility of forming primitive bilayer lipid membranes by long single-chain polyprenyl phosphates in abiotic conditions. It can be the next step in understanding the origin of protocells. PMID- 11014865 TI - Enhanced degradation of hexachlorocyclohexane isomers by Sphingomonas paucimobilis. AB - Hexachlorocyclohexane (HCH) has been banned for use in technologically advanced countries; however, it is still in use in tropical countries like India. Earlier we reported the degradation of HCH isomers by Sphingomonas paucimobilis within 12 days of incubation. Here we report the role of different factors that could enhance the degradation rate of HCH isomers. We found that an increase in the cell number from 10(2) to 10(8) cells/ml resulted in an increased degradation rate of HCH isomers viz. alpha, beta, gamma, and delta-HCH. While alpha-HCH and gamma-HCH disappeared completely from the medium within 3 days of incubation, a maximum of only 90% and 85% degradation was observed for beta and delta-HCH, respectively. We have also observed that adapted cultures degraded HCH isomers more efficiently than did the normal cultures. PMID- 11014867 TI - Comparison of 2,4,6-trinitrotoluene degradation by seven strains of white rot fungi. AB - The degradation of 2,4,6-trinitrotoluene (TNT) by seven strains of white rot fungi was examined in two different media containing 50 mg L(-1) of TNT. When TNT was added into a nutrient-rich YMG medium at the beginning of the incubation, four of the fungal strains completely removed TNT during several days of incubation and showed higher removal rates than those of Phanerochaete chrysosporium. TNT added into YMG medium after a 5-day preincubation period completely disappeared within 12 hours, and the removal rates were higher than those in N-limited minimal medium. Isomers of hydroxylamino-dinitrotoluene were identified as the first detectable metabolites of TNT. These were transformed to amino-dinitrotoluenes, which also disappeared during further incubation from cultures of Irpex lacteus. During the initial phase of TNT degradation by I. lacteus, dinitrotoluenes were also detected after the transient formation of a hydride-Meisenheimer complex, indicating that I. lacteus used two different pathways of TNT degradation simultaneously. PMID- 11014866 TI - Cloning, sequencing, and function of sanF: A gene involved in nikkomycin biosynthesis of Streptomyces ansochromogenes. AB - A 111-bp DNA fragment related to nikkomycin biosynthesis of Streptomyces ansochromogenes 7100 was obtained with the method of reverse genetics. Then, a 2.2-kb DNA fragment was cloned from the DNA library of S. ansochromogenes 7100 by using the 111-bp fragment as a probe. Sequence analysis showed that the fragment contains one complete open reading frame (ORF) that encodes a 219-amino acid (aa) protein, and this gene was designated sanF (GenBank Accession No. AF223971). The function of the sanF gene was studied by a strategy of gene disruption, and the resulting sanF mutants lost the ability to synthesize biologically active nikkomycin, indicating that sanF is essential for nikkomycin biosynthesis. PMID- 11014868 TI - Isolation and characterization of the thylakoid membranes from the NaCl-resistant (NaCl(r)) mutant strain of the cyanobacterium Anabaena variabilis. AB - NaCl-induced changes in the thylakoid membrane of wild-type Anabaena variabilis and its NaCl(r) mutant strain have been studied. Biochemical characterization of the thylakoid membrane was done by taking its absorption and fluorescence spectra at different wavelength. The thylakoid membranes of both strains were isolated by mechanical disruption of the freeze-dried and lysozyme-treated cells, followed by differential and density gradient centrifugation. The light absorption spectra of the thylakoid membrane showed three and two peaks in NaCl(r) mutant strain and its wild-type counterpart respectively at wavelengths of 400-850 nm. These peaks revealed that the thylakoid membrane contains a large amount of carotenoid and chlorophyll a. Fluorescence emission spectra of thylakoid membrane of NaCl(r) mutant and its wild-type strain at excitation wavelength of 335 nm showed two different peaks, one at 340 nm and the other at 663 nm respectively. The light absorption and fluorescence spectra of the thylakoid membrane also revealed that the membrane contained carotenoid pigment, chlorophyll (Chl) a, and a pigment with an emission peak at 335 nm. The HPLC analysis of the pigments of the thylakoid membrane indicates that the NaCl(r) mutant strain under NaCl stress contained an additional peak for the carotenoid pigment, which was lacking in its wild-type counterpart. The major peak in thylakoid membrane was that of echinenone and beta-carotene. Whereas the polypeptide composition of thylakoid membrane differed in the wild-type and its NaCl(r) mutant strain, no difference in the cell wall protein pattern was observed in both strains. The thylakoid membrane of NaCl(r) mutant strain contained two additional protein bands that were absent in its wild-type counterpart. The thylakoid membrane of the wild-type and its NaCl(r) mutant strain also showed morphological variations under NaCl stress. PMID- 11014869 TI - Temperature elevation regulates iron protoporphyrin IX and hemoglobin binding by Porphyromonas gingivalis. AB - Porphyromonas gingivalis, an obligate anerobe with a growth requirement for iron protoporphyrin IX (FePPIX), is exposed to increased temperatures in the inflamed periodontal pocket. In this study, P. gingivalis was grown in a chemostat at 37 degrees C (control), 39 degrees C, and 41 degrees C, and examined for hemagglutinating (HA) activity, hemoglobin binding and degrading activity, and iron protoporphyrin IX binding. HA activity decreased in cells as the growth temperature increased. Binding of mu-oxo bishaem (dimeric haem), and Fe(II)- and Fe(III)-monomeric forms was increased in 39 degrees C-grown cells but decreased in 41 degrees C-grown cells compared with controls. Cellular hemoglobin binding and degradation decreased with increased growth temperature. The decrease in cellular hemagglutination and hemoglobin degradation occurring with increased growth temperature would limit the potential overproduction of toxic monomeric haem molecules. The increased binding of mu-oxo bishaem and monomeric forms of FePPIX at 39 degrees C may reflect a defense strategy against reactive oxidants and a mechanism of dampening down the inflammatory response to maintain an ecological balance. PMID- 11014870 TI - Effects of thymol on ruminal microorganisms. AB - Thymol (5-methyl-2-isopropylphenol) is a phenolic compound that is used to inhibit oral bacteria. Because little is known regarding the effects of this compound on ruminal microorganisms, the objective of this study was to determine the effects of thymol on growth and lactate production by the ruminal bacteria Streptococcus bovis JB1 and Selenomonas ruminantium HD4. In addition, the effect of thymol on the in vitro fermentation of glucose by mixed ruminal microorganisms was investigated. Neither 45 nor 90 microg/ml of thymol had any significant effect on growth or lactate production by S. bovis JB1, but 180 microg/ml of thymol completely inhibited growth and lactate production. In the case of S. ruminantium HD4, 45 microg/ml of thymol had little effect on growth and lactate production; however, 90 microg/ml of thymol completely inhibited growth of S. ruminantium HD4. Thymol also decreased glucose uptake by whole cells of both bacteria. When mixed ruminal microorganisms were incubated in medium that contained glucose, 400 microg/ml of thymol increased final pH and the acetate to propionate ratio and decreased concentrations of methane, acetate, propionate, and lactate. In conclusion, thymol was a potent inhibitor of glucose fermentation by S. bovis JB1 and S. ruminantium HD4. Even though thymol treatment decreased methane and lactate concentrations and increased final pH in mixed ruminal microorganism fermentations of glucose, concentrations of acetate and propionate were also reduced. PMID- 11014871 TI - Evaluation of phospholipid and fatty acid compositions as chemotaxonomic markers of Alteromonas-like proteobacteria. AB - The cellular phospholipids (PLs) and fatty acids (FAs) were investigated in type and environmental strains of Pseudoalteromonas, Alteromonas macleodii, A. infernus, and in three type strains of Marinomonas, M. communis, M. vaga, M. mediterranea. A total of 40 strains (19 strains in this study and 21 reported in previous papers), including Idiomarina abyssalis, I. zobellii, and Glaciecola punicea, G. pallidula, aerobic Alteromonas-like proteobacteria showed genus characteristic patterns of phospholipids and fatty acids useful for genera discrimination. The PL patterns of surface cultures of alteromonads, pseudoalteromonads, and marinomonads consisted almost entirely of phosphatidyl ethanolamine and phosphatidyl glycerol presented in different proportions. Neither diphosphatidyl glycerol nor glycophospholipids were found in bacteria studied. In addition, the minor amount of a glycolipid was found in all strains studied. Bacteria of the genera Marinomonas, Idiomarina, and Glaciecola were clearly distinguished by presence of one of the major FAs: 18:1 (n-7), i15:0, and 16:1 (n-7), respectively. The amounts of these FAs reached up to 40-60% of total FAs. Members of Alteromonas and Pseudoalteromonas were characterized by different ratio of the following major FAs:16:1(n-7), 16:0, 17:1 (n-8), and 18:1 (n-7). PMID- 11014872 TI - Occurrence of multiple ribonucleotide reductase classes in gamma-proteobacteria species. AB - Ribonucleotide reductase (RNR) is central to de novo synthesis of deoxyribonucleotides and essential for all living cells. Three classes have been described; class I is oxygen dependent and represented by two subclasses, Ia (NrdAB) and Ib (NrdEF); class II (NrdJ) is indifferent to oxygen; and class III (NrdDG) is oxygen sensitive. More than one class can be found in an organism, reflecting the oxygen status of its environment. We have investigated, by using PCR and Southern blot, the occurrence of the different classes among species of the gamma-Proteobacteria. Class III are present in all species tested, but the presence of the other classes varies. Some species contain one unique additional enzyme, class Ia, Ib, or II, whereas others contain two additional enzymes, class Ia and Ib, or class Ia and II. PMID- 11014873 TI - Analysis of expression of the binary toxin genes from Bacillus sphaericus in Anabaena and the potential in mosquito control. AB - Anabaena strains expressing the binary toxin genes of Bacillus sphaericus produce high larvicidal activity with living cells. Western blot analysis showed that the 51-kDa and 42-kDa toxin proteins were stable in Anabaena. When a DNA fragment upstream of the 51-kDa protein gene was deleted, the toxicity was reduced by over a hundred-fold, whereas deletions at the coding regions showed that the cooperation of the two proteins expressed in Anabaena is essential for the larvicidal activity. Outdoor tests showed that the genetically altered Anabaena could keep containers with natural water from being inhabited by Culex larvae for over 2 months. PMID- 11014874 TI - Internal glutamine and glutamate pools in Klebsiella pneumoniae grown under different conditions of nitrogen availability. AB - Internal pool sizes of glutamine and glutamate in Klebsiella pneumoniae grown under nitrogen limitation or nitrogen sufficiency were measured to study the signal transduction of external nitrogen limitation. K. pneumoniae cells were grown in an anaerobic, ammonium-limited chemostat culture. At a growth rate of 0.217 h(-1), the steady state ammonium concentration in the culture was 55 microm, correlating with repression of the nitrogen fixation (nif) genes. At growth rates below 0.138 h(-1), the ammonium concentration in the culture dropped below 0.5 microm and the nif genes became derepressed. During the transition from nitrogen sufficiency to nitrogen limitation, the internal glutamine pool in K. pneumoniae decreased by a factor of approximately 6. The glutamate pool, however, remained stable. Similarly, in anaerobic batch cultures with different limiting nitrogen sources, the glutamine pool generally decreased by a factor of 7 to 9 when nif gene derepression was achieved. All the limiting nitrogen sources used resulted in decreased growth rates compared with growth under nitrogen excess, suggesting an inverse relationship between glutamine pool size and doubling time. These studies indicate that K. pneumoniae perceives external nitrogen limitation as internal glutamine limitation. PMID- 11014875 TI - Potassium and sodium fluxes in the phytopathogenic fungus Fusarium oxysporum var. lini. AB - Rubidium uptake in potassium-starved cells followed biphasic kinetics in the micromolar and millimolar range and was independent of the temperature. In contrast, Rb(+) uptake in normal-K(+) cells followed a monophasic kinetics in the millimolar range and increased at temperatures higher than 30 degrees C. Differences in the K(m) values and in the Arrhenius plots of Rb(+) uptake suggest different uptake systems in K(+)-starved and in normal-K(+) cells. In addition, the substantial inhibition of Rb(+) uptake caused by carbonyl cyanide-m chlorophenyl hydrazone indicates that these systems are strongly dependent on membrane voltage. Lithium (sodium) tolerance, influx, and efflux were separately studied. F. oxysporum was shown to be very tolerant to sodium, while lithium caused a specific toxic effect. Li(+) uptake in K(+)-starved cells exhibits a monophasic kinetics with low affinity. Li(+) efflux was not affected by external pH or addition of potassium to the medium, suggesting that a Na(+)/cation antiporter is not involved in this process. PMID- 11014876 TI - Calcium and phosphate regulation of nitrogen metabolism in the cyanobacterium Spirulina platensis under the high light stress. AB - High light stress (40 W/m(2))-induced alterations in the nitrogen assimilatory enzymes in Spirulina platensis were studied under the Ca(2+) and phosphate (Pi) supplemented as well as starved conditions. Results revealed that activities of nitrate reductase (NR), amino acid transferases (AST/GOT and ALT/GPT), and protease enzymes in the high-light-incubated cells were relatively higher under the Ca(2+)- and Pi-starved conditions. On the contrary, relative rates of glutamine synthetase (GS) and ATPase activities were lower in the Ca(2+)- and Pi starved cells. But the Spirulina cells under the Ca(2+)- and Pi-added conditions showed enhanced activity of both GS and ATPase enzymes. During the high-light stress, a decline in the GS activity, particularly under the Ca(2+)- and Pi starved conditions, was indicative of a nitrogen starvation-like condition. This could be one of the reasons for induction of the NR and protease enzymes. A higher rate of GS activity was recorded under both the Ca(2+)- and Pi supplemented conditions, perhaps owing to the enhanced rate of ATPase activity in such conditions. But a declining pattern of both NR and protease activities in the presence of Ca(2+) and Pi, despite the higher rate of ATPase activity, might involve some other mechanism like the protein-kinase system. PMID- 11014877 TI - A comparison of Legionella pneumophila occurrence in hot water tanks and instantaneous devices in domestic, nosocomial, and community environments. AB - The aim of this study was to compare the occurrence of L. pneumophila in hot water samples from hot water tanks and instantaneous devices. Tanks and devices were all operated by heat exchangers employed in the town's district heating system. Thirty-six out of 171 (21%) hot water samples tested positive for L. pneumophila isolation, with 14.6% belonging to serogroup 1 and 6.4% to serogroups 2-14. The proportion of L. pneumophila detected in hot water reservoirs (30%) was higher than that observed in hot water instantaneous devices (6.2%). Differences in L. pneumophila isolation reflected different temperatures registered at the faucet: 60 degrees C for hot water from instantaneous devices. These data emphasize the need to control temperature in hot water distribution devices, thus inhibiting the formation of biofilm and L. pneumophila colonization. PMID- 11014878 TI - Kinematic analysis of laryngeal movements in patients with neurogenic dysphagia before and after swallowing rehabilitation. AB - To examine whether kinematic analysis of laryngeal movements (which are closely linked to pharyngeal swallowing) can differentiate between normal and disturbed swallowing, we used a three-dimensional ultrasound movement recording system to measure the movements of the larynx during swallowing in 32 patients with neurogenic dysphagia caused by central nervous system lesions and in 32 age- and sex-matched healthy individuals. At the beginning of an inpatient rehabilitation swallowing program, laryngeal movements in 24 patients were highly disturbed in terms of velocity curve irregularities. After rehabilitation, the majority of patients with hitherto irregular velocity profiles exhibited laryngeal kinematics that were indistinguishable from those of 32 healthy subjects. Kinematic analysis of laryngeal movements, therefore, is suitable for monitoring motor recovery of swallowing disturbances in patients with neurogenic dysphagia while undergoing swallowing rehabilitation. PMID- 11014879 TI - Effectiveness of four hours of education in interpretation of radiographic studies. AB - Clinicians working with oropharyngeal swallowing disorders often use videofluoroscopy to define their patients' swallowing abnormalities. This study examined the effect of 4 hours of training in the identification of head and neck anatomy and oropharyngeal swallowing disorders viewed radiographically. Ninety clinicians participated in a 5-hour session which included 30-minute pre- and post-tests requiring identification of head and neck anatomy and oropharyngeal swallowing disorders and a 4-hour training period. Results showed significant improvement in identification of both radiographic anatomy and swallowing disorders. The change in pre- and post-test measures was negatively correlated with extent of prior experience in dysphagia. Similar studies are needed with clinicians or students inexperienced in dysphagia to define the number of hours of education needed in order for students to reach a desired accuracy level in their identifications. PMID- 11014880 TI - Prevalence of aspiration and laryngeal penetration in patients with unilateral vocal fold motion impairment. AB - The adverse consequences of aspiration in regard to patient health and quality of life are well documented. It is generally accepted that the probability of aspiration is increased in patients with unilateral vocal fold motion impairment, however, the incidence and proposed mechanism of aspiration vary depending on the reported series. We reviewed the cine or video pharyngoesophagographic findings in patients with documented unilateral vocal fold motion impairment, identified through the Johns Hopkins Hospital Swallowing Center database, to determine the prevalence and cause of aspiration and laryngeal penetration. Aspiration and laryngeal penetration were identified in 38% and 12% of patients, respectively. Aspiration resulted from impaired airway protection, not from esophageal obstruction with laryngeal "spill-over." The number and degree of impairments directly correlated with probability of aspiration. Obstruction potentiated the likelihood of aspiration. Video pharyngoesophagography accurately identifies patients at risk for aspiration. PMID- 11014881 TI - How thick is thick? Multicenter study of the rheological and material property characteristics of mealtime fluids and videofluoroscopy fluids. AB - Objective rheological assessment of fluids given to dysphagic patients at mealtime and during videofluoroscopy was carried out using a multicenter format. Thin, quarter-thick, half-thick and full-thick fluids were examined for the degree of correlation between mealtime fluids and their allegedly matched videofluoroscopy counterparts. The study was carried out to determine whether perceived subjective differences between mealtime fluids and videofluoroscopy fluids could be quantified using the rheological parameters of viscosity, density, and yield stress. The results showed poor correlation between mealtime fluids and videofluoroscopy fluids over all parameters. In general, the videofluoroscopy fluids were more viscous, more dense, and showed higher yield stress values than their mealtime counterparts. Given these results, it is reasonable to assume that the fluids used during videofluoroscopy do not provide an accurate indication of swallowing ability at mealtime. Therefore, it is suggested that clinicians use objective methods to rheologically match videofluoroscopy fluids to mealtime fluids. PMID- 11014882 TI - Use of arterial oxygen saturation, heart rate, and blood pressure as indirect objective physiologic markers to predict aspiration. AB - If an indirect bedside variable can reliably predict whether an objective instrumental dysphagia evaluation is needed, time and money can be saved without compromising patient care. To date, the search for a reliable indirect subjective marker of aspiration has not been successful. However, research on indirect objective markers of aspiration is alluring. The purpose of the present study was to investigate changes, if any, in the physiologic parameters of arterial oxygen saturation (SpO(2)), heart rate, and blood pressure during simultaneous objective confirmation of aspiration status with Fiberoptic Endoscopic Evaluation of Swallowing (FEES). Sixty adult subjects were divided into 4 groups of 15. Group 1 did not require supplemental oxygen and did not aspirate. Group 2 did not require supplemental oxygen and exhibited aspiration. Group 3 required supplemental oxygen and did not aspirate. Group 4 required supplemental oxygen and exhibited aspiration. Simultaneous SpO(2), heart rate, and blood pressure measurements were collected at 1-min intervals, i.e., pre-FEES baseline for 5 min; during FEES; and post-FEES for 5 min. Results indicated no significant differences in SpO(2) levels based on aspiration status or oxygen requirements for any of the 4 groups. A consistent pattern of higher heart rate values during FEES and continuing for 5 min post-FEES was observed for all 4 groups. A consistent pattern of higher blood pressure values during FEES and then lower blood pressure values post-FEES was observed for all 4 groups. It was concluded that the use of changes in SpO(2), heart rate, or blood pressure values as indirect objective markers of aspiration was not supported. PMID- 11014883 TI - Swallowing physiology of toddlers with long-term tracheostomies: a preliminary study. AB - This study investigated the swallowing physiology of toddler-aged patients with long-term tracheostomies. Structural movements and motility of the pharyngeal stage of swallowing were studied in four toddlers ranging in age from 1:2 (years:months) to 2:9 with long-term tracheostomies. A patient aged 1:2 years with no tracheostomy served as a toddler model for comparison. Videofluoroscopic recordings of the patients' liquid and puree bolus swallows were analyzed for a) onset times for pharyngeal stage events, laryngeal vestibule closure, and tracheostomy tube movement; b) timeliness of swallow response initiation; and c) pharyngeal transport function. Results found differences in timing of pharyngeal stage movements between the tracheostomized patients and the patient with no tracheostomy. Laryngeal vestibule closure occurred before or within the same 0.033-s video frame as onset of upper esophageal sphincter (UES) opening in the patient with no tracheostomy, but occurred 0.033-.099 s after onset of UES opening in the tracheostomized patients. The time line required to close the laryngeal vestibule once the arytenoids began their anterior movement was longer in the tracheostomized patients than in the patient with no tracheostomy and was associated with laryngeal penetration. The patient with no tracheostomy displayed superior excursion of the arytenoid and epiglottis during the swallowing; the tracheostomized patients did not. No association was found between onset of tracheostomy tube movement and laryngeal vestibule closure. Delayed swallow response initiation was observed across tracheostomized patients at a mean frequency of 45% with associated penetration. Pharyngeal dysmotility was not observed. Findings supported the concept that long-term tracheostomy in toddler aged patients affects swallowing physiology. PMID- 11014884 TI - Effects of consistent food presentation on oral-motor skill acquisition in children with severe neurological impairment. AB - The purpose of this study was to evaluate systematically the effect of presenting food consistently, in a position regarded as optimal, to children with severe neurological impairment who have associated oral-motor dysfunction. We tested the validity of some recommendations often made in the literature regarding good feeding practices. The trial used an ABA within-subjects design and extended over a 9-month period. Sixteen children between 7 and 17 years of age with severe neurological impairment and associated eating difficulties were studied. Six subjects had some speech. The effects of the intervention were compared by detailed analysis of standard feeding assessments carried out and video-recorded under control and experimental conditions. Statistically significant differences in components of oral-motor behavior were found when a consistent method of food presentation was employed and significant improvements, which could not be attributed to maturation alone, were found between assessment periods. There were also significant differences in the degree of oral-motor learning achieved by children who had some speech and those who had none. Newly acquired skills were not always evident at followup, however, nor in control assessments of feeding. We conclude that some children, even those with severe neurological impairment, can acquire mastery over latent or previously undeveloped oral-motor skills when feeding strategies are modified to allow appropriate opportunities for learning to occur. PMID- 11014885 TI - Regarding "Effects of consistent food presentation on oral-motor skill acquisition in children with severe neurological impairment" (Dysphagia 15:213 223, 2000) PMID- 11014886 TI - "Whole body" mobility after one year of intraoral appliance therapy in children with cerebral palsy and moderate eating impairment. AB - The reciprocal influence of body postures on the oral structures, but also of the oral structures on body postures, has been proposed by clinicians and is taken into consideration when treating children with poor postural control and moderate to severe eating impairments. However, this relationship has not been rigorously investigated. The purpose of this study was to document the possible relationships among oral-motor, postural, and ambulatory control. Ambulatory skills [exclusive use of wheelchair (w/c) vs w/c and ambulation], postural control when sitting, "pathologic" reflexes, and lip and tongue posture were recorded before and after one year of therapy with an intraoral appliance (ISMAR) in 20 children with cerebral palsy and moderate eating impairment. Significant improvement occurred in sitting (head-trunk-foot control) following one year of ISMAR therapy. Ambulatory status also significantly improved above the level of maturation. Half of the children showed marked improvement in oral posture, i.e., their resting mouth posture was closed rather than open. These results support an hypothesis of interaction between oral structures and postural control of the "whole body." Further studies are needed to determine the controls of such a relationship. PMID- 11014887 TI - Assessment of ingestive and oral praxis skills: children with cerebral palsy vs. controls. AB - Eating impairments (dysphagia) in children with cerebral palsy (CP) have been well documented. However, individual components of ingestion, such as the feeding skills of eating and drinking and their relationship to oral-motor planning skills (praxis), remain largely undetermined. The purpose of the present study was to examine functional feeding and oral praxis skills in a group of children with CP and mild eating impairment and to compare their skills with a group of age-matched controls. As well, interobserver reliabilities and concurrent validity of these tests were examined to determine their reliability and to what extent they may be measuring similar constructs. Twenty-seven children with CP and 21 age-matched controls, aged 4.0-16 years, participated in this study. Two oral praxis tests (OFMF and OPT) and two standard ingestive skills tests (GVA and FFAm) were administered to each child. Children with CP scored consistently and significantly lower on the OFMF than controls (p < 0.001); similar results were achieved on the OPT (p < 0.001). Children with CP had difficulty with items where there was a high demand for repetition and smooth sequencing. On the ingestive skills tests children with CP took significantly longer for chewing a hard solid food texture than controls (p < 0.001), and functional feeding skills were also significantly poorer than in controls. Interrater reliability coefficients for the OPT and OFMF were excellent (all ICCs > 0.90). There was high concurrent validity between the OPT and OFMF (r = 0.90, p < 0.0001). The correlations for the FFAm and GVA tests were somewhat weaker (r = -0.54, p < 0. 0001). Correlations were negative; as functional feeding scores increased (improved), chewing time decreased (improved). Excellent reliable and valid assessment instruments are available to the practicing clinician. They cover a wide range of oral-motor performance and, so, must be used judiciously and for the purpose that they have been developed. PMID- 11014889 TI - A comment on "Prevalence of aspiration and laryngeal penetration in patients with unilateral vocal fold motion impairment" (Dysphagia 15:184-187, 2000). PMID- 11014891 TI - Neutropenia accompanying parvovirus B19 infection after gynecologic surgery. AB - The hematologic data and symptoms of 7 patients seropositive for parvovirus B19 IgM antibody after gynecologic surgery were analysed. Parvovirus may have been transmitted by fibrin glue prepared from heat-treated human plasma and used for hemostasis during surgery. The peripheral blood neutrophil count decreased to below 1 x 10(9)/l between postoperative day (POD) 10 and 18, but recovered spontaneously to within the normal range. G-CSF injection was effective in preventing neutropenia or obtaining a prompt recovery. The reticulocyte count fell below 10 x 10(9)/l between POD 13 and 19, and also recovered spontaneously. A slapped-cheek rash was not observed in any of the 7 patients. PMID- 11014890 TI - Expression of wild-type p53 and Bcl-2 family genes oscillates with recurrent remission and relapse in an unusual case of low-grade lymphoma. AB - Downregulation of apoptosis has been proposed as a mechanism of clonal expansion in low-grade B cell neoplasms. We have previously described an unusual case of CD5+ B cell lymphoma characterized by cycles of leukemic phase alternating with spontaneous remission. In the present study, we examined the involvement of apoptosis-related proteins in the progression of this cyclic lymphoma ex vivo. During the leukemic phases, the clonal cells were activated blasts expressing elevated levels of wild-type (wt) p53, Bcl-2, Bcl-x(L), and Bax, while Bak expression increased during the decline of lymphocytosis. Bax heterodimerized with Bcl-2 but not with Bcl-x(L). The anti-apoptotic Bcl-2/Bax heterodimers peaked during early leukemic phases and declined during regression. The elevation in Bcl-2, Bcl-x(L) and Bax expression during early leukemic phases seems to result from cell activation since a similar increase was induced by activating the remission phase leukemic cells in culture. The data suggest that wt p53, Bcl x(L), and Bcl-2/Bax heterodimers support the accumulation of activated leukemic cells during the leukemic phases, while Bax and Bak may be involved in their decline during regression. PMID- 11014892 TI - Prevalence of the C677T methylenetetra- hydrofolate reductase mutation in Thai patients with deep vein thrombosis. AB - We investigated the prevalence of a genetic variation in the 5, 10 methylenetetrahydrofolate reductase (MTHFR) gene (C677T) using polymerase chain reaction techniques in a sample of 500 general Thai population and among 40 unselected Thai patients with an objectively confirmed history of deep vein thrombosis (DVT). The prevalence of the mutated homozygous and heterozygous C677T MTHFR genotype in the group of 500 healthy Thai population was 1.4 and 25.6%, respectively (allele frequency of 14.2%). Of the 40 patients studied, none were homozygotes and 15% were heterozygotes for the C677T MTHFR gene mutation (allele frequency of 7.5%). There was no significant difference in genotype frequency between patients and control groups (p = 0.09). Odds ratios for the probability of the C677T MTHFR gene mutation in the patient versus control group were 0.49 (95% CI 0. 21-1.12). These data indicated that the C677T MTHF gene mutation was not associated with DVT in the Thai population. The lower frequency of the C677T MTHFR gene mutation in our Thai population compared with reports from other studies suggests a wide heterogeneity in the 677T MTHFR genotype frequencies of the different ethnic populations even among Asians. PMID- 11014893 TI - Th1 and Th2 cytokine profiles in sickle cell disease. AB - We have investigated the levels of Th1 (IL-2 and IFN-gamma) and Th2 (IL-4) cytokines in the plasma and supernatants following peripheral blood mononuclear cell culture and mitogen stimulation in a group of 39 patients with sickle cell disease (SCD) made up of 29 SS, 8 Sbeta-thal and 2 Hb SD in steady state. Five SS patients were studied during 7 episodes of vaso-occlusive crisis. Twenty-four control (3 Hb AS and 21 Hb AA) were also studied; 10 were acutely ill while 14 were healthy at the time of the study. The plasma levels of IL-2 and IFN-gamma were similar in the patients and the controls. However, plasma IL-4 was significantly higher among the steady-state SS patients than in the controls. While there was no significant difference in cytokine levels following mitogen stimulation in the different groups, plasma IL-2 to IL-4 and IFN-gamma to IL-4 ratios were significantly lower among the steady-state SS patients, indicating a possible Th2 bias in our sickle cell patients and suggesting a possible mechanism to explain the predisposition of SCD patients to bacterial infections. However, SS patients with good splenic function showed a relative Th1 bias, which may be an additional explanation for the protection against bacterial infections in such patients. PMID- 11014895 TI - Trisomy 14 with thrombocytosis and monocytosis. AB - It has been reported that trisomy 14 is associated with myeloid malignancies, but a case with increased platelet count has also been reported. However, the clinical significance of trisomy 14 is still uncertain. We report a patient with trisomy 14 with thrombocytosis and a gradual increase in monocytosis. He was treated with hydroxyurea, cytarabine and aclarubicin in low doses and his quality of life was maintained for a period of about 1 year from blastic crisis. Hydroxyurea, cytarabine or aclarubicin in low doses may be the treatment of choice for trisomy 14 patients with respect to the patients' quality of life. PMID- 11014894 TI - New imaging findings in a patient with central nervous system dysfunction after bone marrow transplantation. AB - Central nervous system disorders are an important complication of bone marrow transplantation (BMT). We have recently performed cerebral angiography to examine central nervous system dysfunction in a 22-year-old woman with acute lymphoblastic leukaemia who had undergone BMT. Angiography demonstrated multiple stenoses and occlusions in the peripheral branches of the anterior and middle cerebral arteries, a pattern similar to that seen in vasculitis. She was thought to most likely have cytomegalovirus (CMV) vasculitis, but other forms of vasculitis, such as angiitis-like-syndrome-associated graft-versus-host disease could not be excluded. This case suggests that CMV vasculitis may cause central nervous system dysfunction after BMT and that imaging studies may provide useful information about central nervous system disorders in these patients. PMID- 11014896 TI - CD5+ immunophenotype in the bone marrow but not in the peripheral blood in a patient with hairy cell leukemia. AB - We report the case of a patient with rare CD5+ hairy cell leukemia, which was unusual, as there was also a discrepancy between the bone marrow and peripheral blood immunophenotypes. We propose that some degree of maturation within the malignant clone and the predominance of more mature CD5- hairy cells in the peripheral blood can explain this observation. The patient was treated with a conventional course of cladribine (2'-chlorodeoxyadenosine) and achieved complete remission. We conclude that the unusual CD5+ immunophenotype of this patient did not affect prognosis or change the response to standard therapy. PMID- 11014897 TI - ALK-negative anaplastic large cell lymphoma and Hodgkin's disease of nodular sclerosing type in 2 siblings. AB - Familial clustering of Hodgkin's disease (HD) and increased risk of developing the disease among the siblings of affected patients suggest that both environmental and genetic factors may play an important role in its pathogenesis. An association between Epstein-Barr virus (EBV) and HD has been widely demonstrated. Recently, latent membrane protein of EBV has also been detected in CD30-positive anaplastic large cell lymphoma. Familial aggregation of HD and a three- to seven-fold-increased risk among the siblings of affected patients suggest increased genetically determined susceptibility. No data about genetic factors are available for anaplastic large-cell lymphoma. In this study, the authors report the case of a woman with anaplastic-lymphoma-kinase (ALK)-negative CD30-positive anaplastic large cell lymphoma, whose brother had developed HD 11 years previously. The clinical, histologic, and immunohistochemical features of the 2 lymphomas were studied. Both siblings showed bulky mediastinal involvement, effacement of normal lymph node architecture by large, atypical cells, resembling Reed-Sternberg cells, expression of EBV latent membrane protein-1 in the lymph node specimens, concordance of both HLA classes I and II. The clinical presentations and immunological studies disclose numerous similarities between the 2 cases and can suggest that their association is not fortuitous. At present, in problematic cases, a combination of morphologic, immunophenotypic and genetic studies may contribute to better define the tumour type. PMID- 11014898 TI - Acute promyelocytic leukemia with marrow fibrosis at initial presentation: possible involvement of transforming growth factor-beta(1). AB - Although the occurrence of marrow fibrosis in acute myeloid leukemia has been described, there have been no reports of acute promyelocytic leukemia (APL) associated with marrow fibrosis. Here we describe an APL patient with severe marrow fibrosis at initial presentation. He had the typical manifestations of APL, except for marrow fibrosis. Complete remission was achieved by treatment with all-trans retinoic acid plus chemotherapy, and his marrow fibrosis gradually improved concomitantly with the decrease in leukemic cells. To clarify the mechanism of marrow fibrosis in this patient, we investigated the expression of genes for several cytokines promoting fibrosis by the reverse transcriptase polymerase chain reaction methods. An overexpression of transforming growth factor-beta(1) was noted in his leukemic cells at initial presentation, whereas no increase in expression was observed at the time of relapse when he no longer had marrow fibrosis. These findings suggest that overexpression of transforming growth factor-beta(1) was involved in the development of marrow fibrosis in this APL patient. PMID- 11014899 TI - Plasmodium falciparum malaria infection complicated by haemophagocytic syndrome in an old man. PMID- 11014900 TI - Complete blood cell counts in capillary and venous blood of healthy term newborns. PMID- 11014901 TI - wt1 gene expression in childhood acute leukemias. PMID- 11014902 TI - Socio-economic and behavioural risk factors for tooth loss from age 18 to 26 among participants in the Dunedin Multidisciplinary Health and Development Study. AB - OBJECTIVE: To determine the risk factors associated with tooth loss between the ages of 18 and 26. METHODS: Dental examinations at ages 18 and 26 were conducted on Study members in the Dunedin Multidisciplinary Health and Development Study, and sociodemographic and dental service use data were collected using a self report questionnaire. At age 15, an estimate of socio-economic status (SES) for each Study member had been obtained by classifying the occupation of the male parent. A case of tooth loss was defined as an individual who had lost one or more teeth (excluding third molars) due to caries between ages 18 and 26. Logistic regression and Poisson analysis were used to model the occurrence of tooth loss. RESULTS: Among the 821 study members who were examined at both ages, one or more teeth were lost because of caries by 85 (10.3%). After controlling for sex, SES and visiting pattern, baseline caries experience predicted subsequent tooth loss, with the odds increasing by 2.8 for every increase by 1 in the number of decayed surfaces present at age 18. Episodic dental visitors had 3.1 times the odds of their routine visiting counterparts of losing a tooth over the observation period. The number of teeth lost was, on average, 2.3 times higher among episodic dental visitors. CONCLUSIONS: Socio-economic inequalities in tooth loss appear to begin early in the life course, and are modified by individuals' SES and dental visiting patterns. PMID- 11014903 TI - Twenty-four month coronal caries incidence: the role of dental care and race. AB - OBJECTIVES: To describe for a diverse sample of dentate middle-aged and older adults: (1) the 24-month incidence of coronal caries, and (2) its association with a broad range of clinical, behavioral, financial, attitudinal, and sociodemographic factors. METHODS: The Florida Dental Care Study is a prospective observational longitudinal cohort study of 873 persons who at baseline had at least 1 tooth and were 45 years or older. In-person interviews and clinical examinations were conducted at baseline and 24 months, with 6-monthly telephone interviews between those times. A multinomial logistic regression was done to predict whether a participant was in one of four mutually exclusive groups at the 24-month examination (new decay only [NDO]; new filling(s) only [NFO]; both new decay and filling(s) [BOTH]; or neither [NONE]). RESULTS: Only 33% of the 24 month participants were in the NONE group. There was no significant difference in caries incidence between regular attenders and problem-oriented attenders, regardless of whether teeth crowned at baseline, incident crowns, or incident root fragments were excluded. However, once differences in incident tooth loss and baseline clinical, behavioral, financial, and attitudinal differences were taken into account, regular attenders did appear to benefit by developing fewer coronal lesions and fewer dental symptoms than problem-oriented attenders. Baseline carious surfaces, filled surfaces, number of teeth, and bulk restoration fractures predicted caries incidence, but baseline cusp fractures did not. Persons with negative dental attitudes were more likely to be in the NDO and BOTH groups, and negative attitude toward brushing and flossing (but not their frequency) also predicted caries incidence. CONCLUSION: Certain baseline clinical conditions, approach to dental care, ability to pay for dental care, dental attitudes, race, and age group were predictive of coronal caries incidence, and regular attenders appeared to benefit from regular attendance. PMID- 11014904 TI - Caries prediction by multiple salivary mutans streptococcal counts in caries-free children with different levels of fluoride exposure, oral hygiene and sucrose intake. AB - The aims of this study were to assess whether multiple salivary mutans streptococcal (ms) counts have a higher predictive power than a single one, and whether the predictive power of the test is different at different levels of fluoride exposure, oral hygiene and sucrose consumption. Three salivary ms tests were performed at 3-month intervals (positive test: salivary ms >/=5x10(5) cfu/ml) on a sample of 304, initially caries-free, 6- to 7-year-olds. Plaque index was also assessed and the parents filled out a questionnaire concerning sucrose-containing eating events and fluoride exposure. Sensitivity, specificity and Youden's index (J) were used to compare the predictive power. Thirty-six percent of the children developed caries during the 2-year follow-up. The highest J value for a single ms test (the second) was 0.24 (sensitivity 29.1%, specificity 95.4%). The multiple ms test with the highest J value (0.29) was obtained by the dichotomies 0 vs. 1-3 positive tests (sensitivity 50.0%, specificity 79.9%) and 0-1 vs. 2-3 positive tests (sensitivity 31.8%, specificity 97.4%). The predictive power of the multiple test was passable for children with low fluoride exposure and high plaque index (sensitivity 57.4%, specificity 93.7%, J 0.51), while it was low for the other groups. Sucrose exposure did not significantly affect the predictive power of the ms test.sion. PMID- 11014905 TI - In situ acid resistance of in vivo formed white spot lesions. AB - Thin sections of natural enamel lesions, so-called white spots (WS), and areas of sound enamel (SEn) adjacent to the WS were exposed to an intraoral environment for 2 weeks. Thin sections of WS samples, clamped in a PMMA holder, were microradiographed before and after exposure to intraoral conditions. Acid resistance was evaluated by lesion depth and mineral changes during the cariogenic challenge. The results show that there were statistically significant differences in lesion depth, mineral loss and mineral volume percent at the surface before and after the intraoral cariogenic challenge at least at p<0.05, except for a change in mineral volume percent at the surface of WS samples. This exception indicates that no mineral change occurred in the surface layer of WS. The fact of 2.8 and 1.8 times higher ratios of SEn over WS of mean changes in lesion depth and mineral loss data, respectively, seems to indicate a quantitative difference in acid resistance level of WS lesions compared with the areas of SEn. Regarding the site of mineral changes, a distinctive feature of WS samples is that mineral loss occurs at the bottom of lesions. In contrast, areas of SEn produce a typical subsurface type of lesions. From this in situ study, it can be concluded that the surface of WS samples was apparently much more acid resistant (at least approximately 2 times) than the areas of SEn that received a similar intraoral acid challenge. PMID- 11014906 TI - Relationship between mineral distributions in dentine lesions and subsequent remineralization in vitro. AB - Though the mineral distribution of the dentine carious lesion varies largely from tooth to tooth and from patient to patient, there are two main distribution profiles that characterize natural carious lesions in dentine. These profiles include softened and subsurface lesion types. The mineral distribution relationship between the starting profile and the profile after remineralization is not known. In order to study the relational aspects, we have produced demineralized dentine samples in vitro with mineral profiles similar to those of typical natural carious lesions, and subsequently remineralized the samples in a remineralizing solution with various fluoride concentrations (0, 2 and 10 ppm F). The mineral distributions were obtained by using an improved microradiographic technique. In addition, the nature of deposited mineral was analyzed by diamond coupled total internal reflectance spectroscopy. Definite relationship was observed between the original lesion mineral distribution and the mineral distributions following remineralization. The amount of mineral present in approximately the first 50 microm of the lesion influenced the overall mineral profile after remineralization, possibly through influencing ion transport. If the amount was high (> approximately 10 vol%), the deposited mineral was confined to the surface (0-50 microm). The original mineral at those depths acted like a nucleus of mineral regrowth when the amount of residual mineral was intermediate, and like a transport barrier when the surface layer was well mineralized. If a surface barrier was not present, mineral was deposited at deeper depths in the lesion. Fluoride effect on dentine remineralization was dependent on the original mineral content and its distribution in the lesion. Although a high concentration of fluoride was very effective in low-mineral lesions, it produced hyperremineralization on well-mineralized subsurface lesions so that it prevented effective remineralization especially in deeper lesions. PMID- 11014908 TI - Differences in acidogenicity of S. sobrinus and S. rattus are linked to the catalytic efficiency of the glycolytic key enzyme phosphofructokinase. AB - This contribution describes the biochemical properties of two catalytically different phosphofructokinases (PFKs) purified from Streptococcus rattus LB 2 (PFK-rat) and Streptococcus sobrinus OMZ 65 (PFK-sob), respectively. Steady-state kinetics revealed K(M) = 0. 8 mM for PFK-rat and K(M) = 0.08 mM for PFK-sob for F 6-P as the substrate. The enzymes also differ in their pH profiles: whereas the highest activity of PFK-rat was measured at pH = 8.0, the optimum pH of PFK-sob was at pH = 7.0. In addition, compared to PFK-sob, PFK-rat was more sensitive against the allosteric inhibitor ATP. PFK catalyzes a committed step of glycolysis, the main acid producing catabolic pathway. Thus, the catalytically more efficient enzyme isolated from S. sobrinus OMZ 65, especially at low pH, could explain the comparably high acidogenicity of this strain. PMID- 11014907 TI - Fluoride in plaque fluid, plaque, and saliva measured for 2 hours after a sodium fluoride monofluorophosphate rinse. AB - Sodium monofluorophosphate (NaMFP) and sodium fluoride (NaF) are the two most common sources of fluoride used in currently marketed fluoride dentifrices. The purpose of this study was to investigate the effect of mouth rinses containing NaF or NaMFP on the concentrations of fluoride, or the MFP ion, in saliva, whole plaque, and plaque fluid. Twelve subjects abstained from tooth brushing for 48 h, fasted overnight, and then rinsed 1 min with 12 mmol/l (228 ppm [microg/g] F) NaF or NaMFP in the morning. Before the rinse and at 30, 60 and 120 min afterwards, upper and lower molar and premolar plaque samples and whole saliva samples were collected. Aliquots of plaque fluid and centrifuged saliva were obtained from these samples, and the whole plaque residue acid extracted. The F and MFP concentrations were then measured in these samples using ultramicro methods. For both rinses, a higher concentration of plaque fluid fluoride was found at lower molar sites while the reverse was true for the whole plaque fluoride. Furthermore, for both rinses, plaque fluid, whole plaque, but not salivary, fluoride concentrations were above baseline at 120 min. Following the NaMFP rinse, a substantial amount of unhydrolyzed MFP was found in plaque fluid and saliva. Although there was a very large range in these measurements, fluoride in plaque fluid (excluding fluoride in unhydrolyzed MFP) and whole plaque were significantly (p<0.05) greater after the NaF rinse at all time periods. In saliva, the NaF rinse produced a statistically significant greater salivary fluoride (excluding fluoride in unhydrolyzed MFP) only at 60 min. The lack of a clear correlation between these measurements and clinical studies suggest a novel mechanism may enhance the effectiveness of NaMFP dentifrices. PMID- 11014909 TI - Effects of Apis mellifera propolis on the activities of streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite. AB - Propolis, a resinous hive product collected by Apis mellifera bees, has been used for thousands of years in folk medicine. Ethanolic extracts of propolis (EEP) have been shown to inhibit the activity of a mixture of crude glucosyltransferase (Gtf) enzymes in solution. These enzymes synthesize glucans from sucrose, which are important for the formation of pathogenic dental plaque. In the present study, the effects of propolis from two different regions of Brazil on the activity of separate, purified Gtf enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads were evaluated. The EEP from Minas Gerais (MG; Southeastern Brazil) and Rio Grande do Sul (RS; Southern Brazil) were tested for their ability to inhibit the enzymes GtfB (synthesis of insoluble glucan), GtfC (insoluble/soluble glucan) and GtfD (soluble glucan). The effects of propolis on Gtf from Streptococcus sanguis (soluble glucan synthesis) was also explored. The EEP from both regions effectively inhibited the activity of all Gtfs in solution (75-95%) and on the surface of sHA beads (45-95%) at concentrations between 0.75 and 3.0 mg of propolis/ml. However, the two samples of propolis showed different levels of inhibition on each of the enzymes tested. In general, EEP RS demonstrated a significantly higher inhibitory activity on GtfB and C activities (both solution and surface assays) than EEP MG at concentrations between 0.047 and 0.187 mg/ml (p<0.05). EEP MG, on the other hand, exhibited a greater inhibitory effect on the activities of surface GtfD (at 0.375, 0.75 and 1.5 mg/ml) and S. sanguis Gtf (at 1.5 and 3.0 mg/ml; p<0.05). These data indicate that EEP is a potent inhibitor of Gtf enzymes in solution and adsorbed on an experimental pellicle; however, its effect on Gtf activity is variable depending on the geographical origin of the propolis samples. There is a need to identify the active compounds of propolis. PMID- 11014910 TI - Binding characteristics of Streptococcus mutans for calcium and casein phosphopeptide. AB - Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP) and may be used to localize ACP in dental plaque, maintaining a state of supersaturation with respect to tooth enamel, reducing demineralization and enhancing remineralization [Reynolds, J Dent Res 1997;76:1587-1595]. The aim of this paper is to investigate these effects by measuring the affinity and capacity of Streptococcus mutans for CPP-ACP. Using the equilibrium dialysis system described by Rose and Hogg [Biochim Biophys Acta 1995;1245:94-98], assessment of calcium binding by a plaque streptococcus at a fixed CPP-ACP concentration gives a series of CPP-ACP-influenced dissociation constants for calcium. These data can then be used to derive a true dissociation constant for CPP-ACP itself. The results demonstrate that CPP-ACP binds with about twice the affinity of the bacterial cells for calcium up to a value of 0. 16 g/g wet weight cells. Application of CPP ACP to plaque may cause a transient rise in plaque fluid free calcium which may assist remineralization. Subsequently, CPP-ACP will form a source of readily available calcium to inhibit demineralization. Hence, CPP-ACP binds well to plaque, providing a large calcium reservoir, which is likely to restrict mineral loss during a cariogenic episode and provide a potential source of calcium for subsequent remineralization. Overall, once in place, CPP-ACP will restrict the caries process. PMID- 11014912 TI - Immunohistochemical assessment of collagen types I, III, IV and VI in biopsy samples of the bovine uterine wall collected during the oestrous cycle. AB - Uterine biopsies were collected at cycle days 1 (oestrous), 8, 15 and 19 in six cows. Unfixed cryostat sections were used to immunolocalise collagen types I, III, IV and VI by an indirect FITC method. Collagen I was sparsely found in the endometrium where it formed a fine meshwork of thin fibres directly below the surface epithelium, clearly visible only at cycle days 8 and 15. Collagen III formed the bulk of connective tissue fibres and was arranged in fine aggregates within the superficial endometrial stroma, while in the deeper areas it consisted of many thick fibre bundles. Collagen IV was found in basement membranes underlying all endometrial epithelia. Furthermore, it surrounded smooth muscle cells of blood vessels. A few single fibrils also stained positively within the endometrial stroma, more numerous at cycle days 1 and 19 as compared to days 8 and 15. Collagen VI formed a mesh of fine and pericellularly situated fibrils within the endometrial stroma. The contribution of the collagen types studied to the connective tissue of caruncles, blood vessels, lymph follicles, and myometrium is also reported. The results of the present study indicate that the connective tissue of the bovine uterine wall is composed of different collagen types, which exhibit a characteristic distribution pattern each. The day of cycle may influence amounts and organisation of collagen types I and IV as demonstrated here at the light-microscopical level. PMID- 11014911 TI - A randomised controlled trial of the caries-preventive efficacy of a chlorhexidine-containing varnish in high-caries-risk adolescents. AB - A professionally applied two-stage chlorhexidine varnish, Chlorzoin((R)), was developed to achieve sustained release and minimise the problems of staining and bad taste associated with chlorhexidine mouthrinses. The primary aim of this randomised controlled clinical trial was to assess the efficacy of Chlorzoin in reducing the caries increment in high-caries-risk adolescents. Secondary aims included investigating the effect of compliance upon caries increment, the effect of Chlorzoin upon salivary mutans streptococci levels and assessing the benefit of individual dental health advice by dental auxiliaries in a community setting. 1,240 children, initially aged 11-13 years, assessed to be at high caries risk were recruited into the trial. The trial design involved four arms: an observational group, a control group, an active (Chlorzoin) varnish group and a placebo varnish group. All subjects were examined annually by a calibrated examiner who was blind to the group allocation. Three-year caries increments were calculated using clinical, clinical and fibre-optic transillumination, and clinical and bitewing data sets. The results indicated that the use of Chlorzoin had an initial effect on mutans streptococci levels but that no long-term reduction in caries increment or mutans streptococci infection could be detected. One reason for this lack of efficacy may have been the regimen of reduced frequency of varnish applications after the initial period. Children who followed the protocol and, therefore, were seen regularly by dental auxiliaries had a lower caries increment than those who did not. This finding was independent of varnish allocation. In summary, under this regimen, Chlorzoin has been found to be effective in decreasing salivary mutans streptococci but ineffective as a caries-preventive agent in high-risk Scottish children when applied pragmatically in a community setting. PMID- 11014913 TI - Investigation of the female canine urethral vascular plexus using light and scanning electron microscopy: a contributing factor to urinary continence. AB - The vascular system of the female canine urethra was investigated by means of serial histological sections, vascular corrosion casts and scanning electron microscopy. The urethral vascular plexus, located in the proprial connective tissue between the epithelium and the smooth musculature, consisted of sinusoids. It could be divided distally into two parts defined by a constriction in the transitory region between the third and fourth urethral quarters. The proximal segment of the plexus was composed of interconnected longitudinal tubes, whereas the distal part assumed a more net-like form and was continuous with the vestibular plexus. Small arteries usually ran alongside the sinusoidal vessels for a considerable distance before emptying either directly or through branches into the plexus. This vascular arrangement establishes the vascular plexus as a structure similar to an erectile tissue with an arterial inflow, thereby emphasizing the importance of the plexus as a major contributory component guaranteeing urinary continence. PMID- 11014914 TI - Quantitative three-dimensional analysis of chondrocytic kinetic responses to short-term stapling of the rat proximal tibial growth plate. AB - Although it has been demonstrated clinically that controlled compression across a growth plate will slow the rate of endochondral ossification and thus can be used to correct angular limb deformities, the cellular-based mechanism by which altered growth is achieved is poorly understood. This study used short-term uniaxial stapling of the rat proximal tibial growth plate as an experimental system to study chondrocytic responses in the growth plate that account quantitatively for the decreased rate of growth. Growth plates were labeled with oxytetracycline to measure bone growth, and with bromodeoxyuridine to analyze proliferative cell kinetics. Multiple indicators of chondrocytic activity, measured by stereological parameters, were analyzed using growth rate as the primary dependent variable. The unique feature of this analysis was the creation of three-dimensional reconstructions that allowed analysis of data in all directions with distance from the staple. A significant observation was that for the entire operated limb after both 3 and 6 days, all chondrocytic kinetic parameters were affected, indicating that proliferative and hypertrophic responses both act to decrease growth rate in response to stapling. This contradicted our hypothesis that proliferative and hypertrophic responses could occur independently, and that small changes in rate would be attributed primarily to the former and large changes to the latter. The data from this study also demonstrate that volume regulation during hypertrophy can be affected by a primarily mechanical perturbation. Because changes in hypertrophic cell number and volume throughout the growth plate that occur by day 3 remain similar at day 6, the initial modulation of chondrocytic volume and shape may represent the limit of the response while maintaining a growth plate capable of continued growth. PMID- 11014915 TI - The superficial anulus fibrosus ligament. An incipient description of a separate ligament between the lumbar anterior longitudinal ligament and the intervertebral disc. AB - A previously unknown ligament, the superficial anulus fibrosus ligament (SAFL), situated on the ventral part of the L5 intervertebral disc (ID) was observed and described from autopsy material. Twenty-eight cadaveric specimens from 12 black and 16 white persons aged 17-30 years were studied during routine forensic autopsies. The anterior longitudinal ligament was separated from the ID and the ventral part of the SAFL was visualized. The SAFL samples were removed, measured and studied with both conventional histology and examination by transmission electron microscopy. The SAFL was a completely separate ligament at the level of the L5-S1 ID situated between the anterior longitudinal ligament and the anulus fibrosus of the ID. The fibers of the ligament were vertically oriented. A difference in distance between the L5-S1 vertebral bodies ventrally was noted in the two groups studied (18.7 +/- 1.2 mm in the black vs. 15.2 +/- 1.0 mm in the white, p < 0.001), indicating a difference in the ventral thickness of the intervertebral disc. Also, there was a difference in the length (black: 17.7 +/- 1.6 mm vs. white: 14.1 +/- 1.1), thickness (black: 3.3 +/- 0.3 mm vs. white: 2.1 +/- 1.9), and the cross-sectional area (black: 58.2 +/- 6.7 mm(2) vs. white: 26.5 +/- 2.7 mm(2), p < 0.001) of the SAFL. Conventional light microscopy revealed no obvious differences. However, transmission electron microscopy revealed notably larger collagen fibril diameter in black than white subjects. In conclusion, the interbody distances were greater in the black group, indicating a greater intervertebral disc thickness, compared to that of the white. Furthermore, the SAFL was significantly longer and thicker in the black than in the white group. Albeit unsubstantiated, these race-specific macroscopic findings may have implications for understanding the etiology of various low back stress problems. PMID- 11014916 TI - Changes in the structure of dentine from cheek teeth of deer chronically exposed to high levels of environmental fluoride. AB - The purpose of this study was to examine, using backscattered electron imaging, the changes occurring in the structure of the dentine including giant tubules in the teeth of deer chronically exposed to fluoride. The primary and secondary dentine was characterised by the presence of interglobular dentine and regular bands of hypo- and hypermineralised dentine. Giant tubules in the unworn teeth of animals exposed to low and high levels of fluoride were characterised by a large lumen and hypermineralised rim in which there were calcospherites. With the occlusal wear of the teeth tubules beneath the immediate surface of the exposed dentine in teeth from fluorotic animals became occluded with mineral, but in low fluoride animals some tubules at the surface were not occluded with mineral. It is suggested that mineral which occludes the tubules may come from both intrinsic and extrinsic sources. The presence of some open tubules at the immediate surface in the worn teeth of animals exposed to low fluoride suggests an extrinsic source for this mineral. PMID- 11014917 TI - The dentinal structure of equine incisors: a light and scanning electron microscopic study. AB - This paper gives an anatomical overview of the dentinal structure in equine incisor teeth with special reference to the three-dimensional organization, the number and the diameter of the dentinal tubules. The spatial arrangement of equine dentine was examined by scanning electron microscopy of occlusal surfaces and longitudinally fractured teeth and by light microscopy of both decalcified and ground sections. The dentinal tubules of the peripherally situated primary dentine were directly continuous with those of the circumpulpal secondary dentine. The tubules had numerous side branches along their entire course and ramified into terminal branches near the dentino-enamel junction. Tubules of tertiary dentine, situated in the centre of the dental star, were few in number and not continuous with those of the surrounding secondary dentine. On non-etched incisors tubular orifices were clogged by an amorphous smear layer that covered the occlusal surface. On etched occlusal surfaces the numerical tubular density was calculated in different zones. The largest number of tubules per unit area was situated in the secondary dentine forming the dark periphery of the dental star, whereas the smallest number was present in the tertiary dentine located in the pale centre of the dental star. Dentinal tubular diameters were measured at various distances from the pulp. They were widest near the pulpal wall and narrowed progressively towards the dentino-enamel junction. Tubular widths measured on etched occlusal surfaces were considerably larger than those measured on undecalcified incisors due to the removal of intratubular dentine during the etching process. PMID- 11014918 TI - Delayed expression of calbindin D28k during regeneration of the periodontal Ruffini endings of the rat incisor following injury to the inferior alveolar nerve. AB - Expression of calbindin D28k (CB)-like immunoreactivity (-LI) was compared with that of protein gene product 9.5 (PGP 9.5), a general neuronal marker, in the periodontal ligament of the rat lower incisor following resection of the inferior alveolar nerve (IAN). In normal animals, the periodontal nerve fibers showing PGP 9.5-LI formed either Ruffini endings with expanded arborization or thin free nerve endings in the alveolar half of the ligament. Thick CB-like immunoreactive (-IR) nerve fibers terminated in a dendritic fashion in the same region, but thin CB-IR nerve fibers were rarely detected. During the 3 days following resection of the IAN, most of the PGP 9.5-IR and all CB-IR nerve fibers disappeared. Regenerated PGP 9.5-IR nerve fibers appeared around 7 days after resection, in contrast to the very small number of regenerated CB-IR nerve fibers. Around 21-28 days following resection, the number and terminal morphology of regenerated PGP 9.5-IR nerve fibers were comparable to those observed in normal animals, but the number of regenerated CB-IR nerve fibers was still smaller. The terminal morphologies of these regenerated CB-IR nerve fibers showed less expansion compared with normal animals at these post-injured periods. The number of regenerated CB-IR nerve fibers increased gradually to return to normal by 56 days following injury. The delayed expression of CB in the regenerated periodontal Ruffini endings suggests that the functional recovery of periodontal Ruffini endings occurred after the regeneration of periodontal Ruffini endings had been completed. PMID- 11014919 TI - Structure of the middle ear and auditory tube in the house musk shrew, Suncus murinus. AB - Anatomical features of the middle ear and auditory tube (AT) in the house musk shrew, Suncus murinus, were examined by dissection and light microscopy. The tensor veli palatini (TVP) and tensor tympani (TT) have no connections with the wall or cartilage of the AT although they are connected by the intermediate tendon. None of the levator veli palatini (LVP) muscle bundles are attached to the AT. The salpingopharyngeus (SA) alone has its origin on the caudal edge of the tubal cartilage. The origin extends to the pharyngeal two thirds of the cartilage. The SA originates perpendicular to the AT and runs caudomedialward. Some SA muscle bundles intermingle with those of the palatopharyngeus to end on the dorsal wall of the pharynx. The observations provide no evidence that the TVP, LVP and TT have any role in AT function. The only muscle affecting the AT function in S. murinus is the SA, and it would be the AT dilator. PMID- 11014920 TI - Localization of P2X receptors in the guinea pig adrenal gland. AB - The distribution of each of the seven subtypes of adenosine 5'-triphosphate (ATP) gated P2X receptors was investigated in the adrenal gland of guinea pig utilizing immunohistochemical techniques. The occurrence of positive immunoreactivity with specific distribution was observed for antibodies against P2X(1), P2X(2), P2X(5) and P2X(6) receptors, whereas no immunoreactivity was found for antibodies against P2X(3), P2X(4) and P2X(7) receptors. Immunoreactivity for P2X(1) occurred in cells of the inner region of the zona reticularis of the cortex, Several P2X(2)-immunoreactive connective tissue-like elements were located between groups of cortical cells of the zona reticularis. Bundles and terminals of nerve fibres as well as intrinsic neurones gave positive immunoreactivity for P2X(5). The immunoreactive nerve fibres appeared to belong to both extrinsic preganglionic sympathetic and intrinsic immunoreactive neurones. Chromaffin cells were immunoreactive for the P2X(6) receptor antibody. The widespread and specific distribution of P2X receptor subtypes in the adrenal gland suggests significant roles for purinergic signalling in the physiology of the guinea pig adrenal gland. PMID- 11014921 TI - Measurement of hepatic tissue hypoxia using near infrared spectroscopy: comparison with hepatic vein oxygen partial pressure. AB - Hepatic hypoxia occurs during liver surgery and transplantation. The critical level associated with irreversible hepatocellular damage is unknown. Measurement of hepatic tissue oxygenation and hepatic vein oxygen partial pressure (HVPO(2)) reflects oxygen supply and consumption. Near infrared spectroscopy (NIRS) can be used to monitor hepatic oxyhaemoglobin (HbO(2)), deoxyhaemoglobin (Hb) and cytochrome oxidase (Cyt Ox) oxidation. This study compared regional hepatic tissue oxygenation (HbO(2), Hb and Cyt Ox) using NIRS with HVPO(2). The use of tissue oxygenation measured by NIRS and HVPO(2) as indicators of hepatic tissue hypoxia was also investigated. Large Landrace pigs (n = 5) underwent laparotomy and liver exposure. Systemic and hepatic haemodynamics were monitored continuously. NIRS probes were placed on the liver to record continuously HbO(2), Hb and Cyt Ox. Graded hypoxaemia was achieved by stepwise reduction of the fraction of inspired oxygen (FiO(2)) from 30% (baseline) to 4%. A significant decrease in hepatic arterial blood flow and total hepatic blood flow was seen with severe hypoxaemia while there was no significant change to portal vein blood flow. Oxygen partial pressures in the hepatic artery, portal vein and hepatic vein decreased progressively with all grades of hypoxaemia. There was an immediate reduction of hepatic HbO(2) and simultaneous increase in hepatic Hb with all grades of hypoxaemia. Hepatic Cyt Ox was reduced significantly only with FiO(2) < or =10%. A significant correlation (p<0.001) was found between the changes in hepatic oxygenation parameters measured by NIRS and HVPO(2). HVPO(2) measurement did not predict the reduction in intracellular tissue oxygenation demonstrated by NIRS with a decrease of Cyt Ox oxidation. In conclusion there was a good correlation between the tissue oxygenation parameters measured by NIRS and HVPO(2). However, the reduction of intracellular oxygenation found with severe hypoxaemia was demonstrated only by NIRS. PMID- 11014922 TI - The effects of allopurinol in hepatic ischemia and reperfusion: experimental study in rats. AB - BACKGROUND/AIMS: Some studies have shown that postischemic hepatic dysfunction is mainly due to oxygen free radicals that are generated by xanthine oxidase. The present study was undertaken to determine the effect of allopurinol, an inhibitor of xanthine oxidase, on oxidative stress, liver injury and histologic alterations induced by hepatic ischemia-reperfusion in rats. METHODS: One hundred and sixty Wistar rats were used and divided into three groups. Group 1: sham operation; group 2: 50 min of ischemia followed by 1 h of reperfusion, and group 3: pretreatment with allopurinol and 50 min of ischemia followed by 1 h of reperfusion. The effect of allopurinol was evaluated by plasma levels of alanine aminotransferase and aspartate aminotransferase, histopathologic studies, and lipid peroxidation measured by the thiobarbituric acid reactive substances method and chemiluminescence initiated by tert-butyl hydroperoxide technique. RESULTS: Ischemia followed by reperfusion promoted an increase in lipid peroxidation of the hepatic cells when compared to the sham-operated group (p<0.05). This increase was attenuated in the group treated with allopurinol (p< 0.05). Allopurinol also showed a protective effect on hepatocellular necrosis (p<0.05), and the plasma levels of liver enzymes returned earlier to the normal range in rats pretreated with allopurinol in comparison to those that did not receive the drug (p<0.05). CONCLUSIONS: Allopurinol exerted a protective effect on hepatic ischemia and reperfusion in rats. The administration of this drug prior to liver operations should be considered to be submitted to trials in humans. PMID- 11014923 TI - Hepatic lymphocyte transplantation in hyperbilirubinemic gunn rats. AB - Hepatic lymphomyeloid cells (HLCs) are thought to contain liver stem cells. We investigated whether HLCs generated enzyme-producing cells in vivo. HLCs from normal Wistar/Shi rats and rats in which liver ischemia was induced using a portal clamp 4 days previously were studied histopathologically and characterized using flow cytometry. Splenic lymphocytes obtained from these animals were compared as a control. The proliferative activity of HLCs and splenic cells from both groups was also tested by stimulation with concanavalin A. HLCs contained a significantly higher number of NK-T cells and OV6+ cells compared with the splenic cells. The HLCs from rats in which liver ischemia was induced tended to have greater proliferative activity than those from normal rats, while the proliferative activity of splenic lymphocytes was impaired by liver ischemia. The HLCs obtained from Wistar/Shi rats with liver ischemia were then injected into hereditary hyperbilirubinemic Gunn rats to determine whether the HLCs generated enzyme-producing cells. After injection of these stimulated HLCs, the titer of serum bilirubin in the recipient rats was markedly reduced over a long time course (6.80+/-0.93 to 4.87+/-0.22 mg/dl after 1 month and 3.52+/-1.33 mg/dl after 6 months). The results of the present study indicate that HLCs have different populations than splenic cells, and ischemia-reperfusion of the liver increased their proliferative activity. HLC transplantation successfully reduced high bilirubin levels over a long time course. PMID- 11014924 TI - Influence of positive end-expiratory pressure ventilation on peripheral tissue perfusion evaluated by measurements of tissue gases and pH. An experimental study in pigs with oleic acid lung injury. . AB - Measurements of subcutaneous oxygen tension (PscO(2)), subcutaneous carbon dioxide tension (PscCO(2)) and subcutaneous pH (pHsc) were used for evaluation of peripheral oxygenation in pigs subjected to oleic acid-induced lung injury during ventilation with increasing levels of positive end-expiratory pressure (PEEP). Lung injury resulted in a decrease of arterial oxygen tension (PaO(2)) from 93 to 37 mm Hg (p<0.01) with maintained cardiac output. PscO(2) decreased from 45 to 17 mm Hg (p<0.01) and pHsc from 7.47 to 7.39 (p<0.05), and PscCO(2) increased from 46 to 59 mm Hg (p<0.05). Increase of PEEP level between 5 and 20 cm H(2)O resulted in a continuous increase of PaO(2) from 45 to 145 mm Hg and a decrease of cardiac output from 4.1 to 2.0 liters/min (p<0.01). PscO(2) increased up to a PEEP level of 15 cm H(2)O, reaching 26 mm Hg. Further increase of PEEP level up to 20 cm H(2)O resulted in an increase of PscCO(2) from 65 to 71 mm Hg (p<0.05) and a decrease of pHsc from 7.31 to 7.29 (p<0.05). IN CONCLUSION: measurements of tissue gases and pH can be used to evaluate optimum peripheral tissue oxygenation during titration of PEEP level. Whether these measurements can be used as the only indicator to guide therapy in an individual case remains to be studied. PMID- 11014925 TI - Intimal hyperplasia after long-term venous catheterization. AB - OBJECTIVE: Intimal hyperplasia is a well-known consequence of arterial injury and arterialization in vein grafts. However, the subacute and chronic vein wall changes which occur after catheterization have not been well studied. In this animal study, intimal hyperplasia in the vein wall after catheterization was examined. METHODS: A silicon catheter was placed in the anterior caval vein of 54 rats. After in situ fixation at scheduled intervals (1 day to 6 months), the pathologic changes in the vein wall were studied on semi-serial histology sections by means of light microscopy. RESULTS: Three forms of intimal hyperplasia could be observed: plaque-like, papillary-like and incorporation of the mural part of the sleeve into the underlying vein wall. Although the appearance of each was different, their composition was identical. All were mainly composed of alpha-actin-positive cells and collagen localized above the internal elastin layer, and covered by endothelium if facing the lumen. The plaque-like and papillary-like forms were mainly localized in the anterior vena cava, while sleeve incorporation mainly occurred in the jugular vein. Plaque-like and papillary-like intimal hyperplasia could be seen together on the same slide, but these two forms were never seen together with sleeve incorporation. CONCLUSION: Intimal hyperplasia occurs after venous catheterization and is probably caused by chronic injury to the vein wall due to knocking and rubbing movements of the catheter against the wall. PMID- 11014926 TI - Diverting colostomy increases anastomotic leakage in the rat colon. AB - In the present study, we examined the effect of a diverting colostomy on the intestinal healing of colonic anastomosis in the rat. For this purpose, we created a colonic stenosis 2 days prior to the formation of a distal one-layer end-to-end anastomosis with or without a proximal double-barreled deviation colostomy in the rats. Radiological examination of anastomotic leakage was performed daily for 4 days and on day 7 after the operation. We found that anastomotic leakage was markedly increased in rats with a diverting colostomy compared to control animals; i.e. the leakage index (percentage of days with leakage during the experimental period) in colostomy rats was 29%, whereas in animals with no colostomy, the leakage index was only 7%. Interestingly, it was observed that anastomosis formation was associated with a higher mortality rate in rats with colostomy diversion (36%) compared to control animals (7%). However, there was no difference in suture holding capacity on day 7. Moreover, body weight decreased significantly in the colostomy group compared to rats without surgical defunctioning when followed for up to 7 days after surgery. Taken together, our novel findings suggest that a diverting colostomy may increase intestinal leakage after anastomosis formation in the rat colon. Thus, the role of proximal colostomy in the protection of colorectal anastomosis needs to be reevaluated and further investigations are required to resolve the influence of surgical defunctioning on intestinal healing. PMID- 11014927 TI - 11th walter brendel symposiumon applied immunology and microcirculation. mauls, Sudtirol, italy, february 7-9, 2000 PMID- 11014928 TI - Disorders of the epithelial Na(+) channel in Liddle's syndrome and autosomal recessive pseudohypoaldosteronism type 1. AB - The epithelial Na(+) channel (ENaC) is the key step in many Na(+)-absorptive epithelia, such as kidney and distal colon, that controls the overall rate of transepithelial Na(+) transport. ENaC is composed of three homologous subunits, alpha, beta, and gamma. The alpha subunit is the key subunit for the formation of a functional ion channel, while the beta and gamma subunits can greatly potentiate the level of expressed Na(+) currents. ENaCs belong to the recently identified DEG/ENaC supergene family, sharing the same basic structure with cytoplasmic amino and carboxy termini, two transmembrane regions, and a large extracellular loop. The human ENaC genes have been cloned, and using genetic linkage analysis the involvement of ENaC gene mutations in two distinct human diseases, Liddle's syndrome and autosomal recessive pseudohypoaldosteronism type 1 (PHA-1), has been demonstrated. In Liddle's syndrome, gain-of-function mutations in the beta or gamma ENaC subunits have been found; all identified mutations so far reside in the carboxy terminus of the protein, either deleting or modifying the functionally important PY motif. In PHA-1, loss-of-function mutations in the alpha, beta, or gamma subunits have been found; these mutations either truncate a significant portion of the structure or modify an amino acid that plays an important role in channel function. In this review, our current understanding about ENaC and the pathophysiology of Liddle's syndrome and PHA-1 caused by ENaC mutations will be discussed. PMID- 11014929 TI - Defective processing and trafficking of water channels in nephrogenic diabetes insipidus. AB - Nephrogenic diabetes insipidus (NDI) is a disease characterized by the inability of the kidney to concentrate urine upon stimulation with vasopressin. Mutations in the gene for aquaporin-2 (AQP2) are the cause of the autosomal recessive and autosomal dominant forms of NDI. Mutant AQP2 proteins, found in autosomal recessive NDI, were shown to be misfolded and retarded in the endoplasmic reticulum. One mutant protein leading to autosomal dominant NDI, E258K, has been analyzed in detail. It was shown that this mutant was not retarded in the endoplasmic reticulum but mainly retained in the Golgi network. Furthermore, this particular mutant was able to form heterotetramers with wild-type AQP2, in contrast to mutants found in autosomal recessive NDI. The subsequent misrouting of complexes containing wild-type and mutant AQP2 proteins explains dominant NDI. PMID- 11014930 TI - Defects in processing and trafficking of cystic fibrosis transmembrane conductance regulator. AB - In most epithelial tissues Cl(-) transport relies on the cystic fibrosis transmembrane conductance regulator (CFTR) which has dual function as a Cl(-) channel and as a regulator of other ion channels. More than 900 different mutations in the CFTR gene are the cause for defective transport of Cl(-) and Na(+) and impaired secretion or absorption of electrolytes in cystic fibrosis. However, the CFTR mutation delta F508 is the most common reason for the frequently inherited disease among the Caucasian population. Maturation and processing of delta F508-CFTR is defective which leads to expression of only very little but functional CFTR in the cell membrane. Understanding the processing and trafficking of CFTR may give a clue to the question as to how the expression and residual function of delta F508-CFTR can be enhanced, and may lead to the development of new pharmacological tools for the treatment of cystic fibrosis. PMID- 11014931 TI - Mechanisms of renal calcium transport. AB - The kidneys play a key role in the integrated regulation of calcium homeostasis. Calcium absorption takes place throughout the nephron. Proximal tubules, thick ascending limbs of Henle's loop, and distal tubules are the major sites of calcium absorption. The mechanisms of absorption vary significantly from one segment to another, as does the extent of hormonal regulation. At one extreme is the considerable reabsorption by proximal tubules that proceeds primarily, if not entirely, by a paracellular pathway that is not regulated by hormones or drugs. In thick ascending limbs, calcium absorption occurs through a combination of transcellular and paracellular routes. The active, transcellular component is regulated by parathyroid hormone (PTH) and calcitonin, whereas the passive, paracellular route is governed by the extent of concomitant sodium absorption. At the other extreme is the distal tubule, where calcium absorption is entirely transcellular and is regulated by PTH,1,25[OH(2)] vitamin D(3), calcitonin, and by calcium-sparing drugs such as thiazide-type diuretics. The present review focuses on recent insights into the mechanisms of transcellular calcium movement and highlights the discovery of an epithelial calcium channel, ECaC, that may mediate calcium entry in distal tubules. PMID- 11014932 TI - Molecular pathology of renal chloride channels in Dent's disease and Bartter's syndrome. AB - Recent advances in molecular biology have characterised a new class of chloride channels that are referred to as voltage-gated chloride channels (CLCs). To date 9 such CLCs (CLC-1 to CLC-7, CLC-Ka and CLC-Kb which are respectively encoded by the genes CLCN1 to CLCN7, CLCNKa and CLCNKb) have been identified in mammals. Mutations in 2 of these, referred to as CLC-5 and CLC-Kb, have been defined in the hypercalciuric nephrolithiasis disorders of Dent's disease and a form of Bartter's syndrome, respectively. In addition, other forms of Bartter's syndrome have been defined with mutations involving the bumetanide-sensitive sodium potassium-chloride co-transporter (NKCC2) and the potassium channel ROMK. Finally, mutations of the thiazide-sensitive sodium chloride co-transporter (NCCT) are associated with Gitelman's syndrome, in which hypocalciuria and hypomagnesaemia are notable features. These molecular genetic studies have increased our understanding of the renal tubular mechanisms that regulate mineral homeostasis. PMID- 11014933 TI - Severely impaired urine-concentrating ability in mice lacking the CLC-K1 chloride channel. AB - To analyze the physiological functions of CLC-K1 in vivo, we generated mice lacking CLC-K1 by targeted gene disruption. Homozygous mutant Clcnk1-/- mice produced approximately 5 times more urine than Clcnk1+/- and Clcnk1+/+ mice. After 24-hour water deprivation, Clcnk1-/- mice became severely dehydrated and lethargic. Intraperitoneal injection of the V2 agonist, deamino-Cys(1), D-Arg(8) vasopressin, induced an increase in urine osmolarity in Clcnk1+/- and Clcnk1+/+ mice from approximately 1,000 to approximately 3,000 mosm/kg H(2)O, whereas the increase in Clcnk1-/- mice was only from approximately 600 to approximately 840 mosm/kg H(2)O, indicating nephrogenic diabetes insipidus in Clcnk1-/- mice. These results clearly established that CLC-K1 plays a major role in the urinary concentrating mechanisms. PMID- 11014935 TI - Changes in Finnish dental consonant articulation in cleft lip/palate children between 6 and 8 years of age. AB - The aim of this study was to clarify changes in the occurrence of misarticulations of dental consonants /r/, /s/ and /l/ between 6 and 8 years of age in cleft-affected Finnish children and to compare the effects of gender and to estimate if spacing due to changing of maxillary incisors could explain changes in articulation of these sounds. The subjects were 133 (47 girls, 86 boys) Finnish-speaking non-syndromic children with isolated cleft palate (n = 34), cleft lip/alveolus (n = 49), unilateral (n = 33), and bilateral (n = 17) cleft lip and palate. The results showed that there were no statistically significant differences between genders according to changes in articulation abilities by the age of 8 years. Eighty-one percent of the subjects with misarticulations at the age of 6 years still misarticulated at 8 years of age, and 16% of the subjects without misarticulations at the age of 6 years misarticulated at the age of 8 years; /s/ and /l/ misarticulations were eliminated more often than /r/ errors. New misarticulations were diagnosed at the age of 8 years in the same way in the groups with (14%) and without (16%) misarticulations at the age of 6 years. Dental arch spacing due to changing of maxillary incisors does not seem to explain new misarticulations of /r/, /s/ and /l/ sounds estimated at the age of 8 years. PMID- 11014934 TI - Molecular characteristics of phosphate transporters and their regulation. AB - A key process in overall P(i)-homeostasis is renal proximal tubular reabsorption of inorganic phosphate (P(i)), which involves secondary active sodium/phosphate (Na(+)/P(i)) cotransport reabsorption at the brush border membrane. Among the two different molecularly identified Na(+)/P(i) cotransporters, the type-IIa Na(+)/P(i) cotransporter (NaPi-IIa) accounts for up to 70% of brush border membrane transport. Regulation of renal P(i) reabsorption centers around brush border membrane insertion and retrieval of transporter protein under the influence of hormonal and nonhormonal factors. Immunohistochemical and fluorescence techniques have provided new insights into the tissue distribution and the regulation processes. The intrinsic electrogenicity of NaPi-IIa, has allowed detailed studies of the transport kinetics of NaPi-IIa and, combined with mutagenesis methods, structure-function information at the protein level is emerging. PMID- 11014936 TI - Developing a language screening test for Arabic-speaking children. AB - This report gives an account of general and specific issues associated with developing an Arabic-language screening test for children aged between 3 and 12 years. Challenges, dilemmas and stages in the developmental process are discussed. The development of the Arabic Language Screening Test: Preschool and School Age illustrates the process. Issues and stages included: (1) arriving at a consensus about scope and purpose; (2) conceptualizing tasks and items; (3) specifying item content and structure; (4) field-testing the screening tool in a circumscribed environment; (5) developing a robust scoring and interpretation system for determining Pass or Fail, and (6) establishing the degree of accuracy in differentiating children with and without language disorders. The product is a screening test of verbal and related nonverbal abilities with parallel components for children of preschool (3-5 years) and elementary school age (6-12 years). Normative data were collected for 750 Arabic-speaking children in Jordan and Palestine, distributed fairly equally between the ages of 3 and 12 years. Normative means increased with age and standard deviations decreased. Interscorer agreement (99%), internal consistency (Cronbach alpha = 0.98) and diagnostic sensitivity (over 90% accuracy) were high. Constraints and limitations in the development of the test are described and discussed from both objective and personal perspectives. PMID- 11014937 TI - A method to measure speaking time and speech sound pressure level. AB - Many voice disorders are associated with vocal loading, but easy measurement of the vocal load has not been possible so far. To fulfil this need, an easily accessible method was developed and tested. Speaking time and speaking sound level can be recorded during normal work, and measurement can be made in a noisy environment. The measurement is made with a pair of modern noise exposure analysers from which the data is transferred to a personal computer, and the voice speaking time and the related sound pressure levels are calculated with a spreadsheet program. Suitable meters are available from various manufacturers. The method may be applied to measure the need of voice use in different professions, assessing a person's performance in his or her work or evaluating the effects of voice therapy on speaking performance. PMID- 11014938 TI - Treatment of reflux-related and non-reflux-related dysphonia with profound gastric acid inhibition. AB - Thirty-eight patients with dysphonia exceeding 3 months, not caused by trauma, infection, paralysis or allergy, were studied. In all patients a 24-hour ambulatory dual-probe pH monitoring, 5 and 20 cm above the lower oesophageal sphincter was performed. Subsequently they were treated with lansoprazole 30 mg once daily during 6 weeks. A voice range profile, perceptual evaluation of the voice and videolaryngostroboscopy were performed in all patients before and after treatment. A questionnaire about laryngeal symptoms and heartburn was completed on the same two occasions. Thirty-nine percent (15 out of 38) of the patients had an abnormal pH profile and were considered to have reflux-related dysphonia. Only these patients showed a significant improvement in their subjective score on dysphonia (p < 0.05), chronic cough (p < 0.05), dysphagia (p < 0. 05) and heartburn (p < 0.01). Also posterior erythema decreased significantly in these patients (p < 0.01). Only 2 of the 15 subjects did not complain of heartburn. PMID- 11014943 TI - Changes in plasma adenosine concentrations during normal pregnancy. AB - The aim of this study was to measure changes in plasma adenosine concentration [ADO] during a normal pregnancy and to evaluate the possible role of platelets and red blood cells (RBC) as causes of changes in plasma [ADO]. We measured the plasma [ADO] in normal pregnant women (n = 11) during the first, second and third trimesters. The mean plasma [ADO] in the third trimester was 0.41 +/- 0.08 microM (means +/- SEM), significantly higher than in the first and second trimesters (p < 0.05). In pregnant women, platelet and RBC counts, hematocrit and hemoglobin concentration decreased slightly throughout the pregnancy. The elevation in the plasma [ADO] correlated inversely with the platelet count (r = -0.43, p < 0.05). These results suggest that an increase in the plasma [ADO] in the third trimester may be attributed to the enhanced adenosine release from activated platelets. PMID- 11014944 TI - Nausea and vomiting in pregnancy: an association between symptoms and maternal prostaglandin E2. AB - This study investigated the aetiology of nausea and vomiting in pregnancy (NVP) in primary care using a new methodology. Eighteen women in early pregnancy had 2 blood samples taken in one 24-hour time period, one when they were symptomatic, and another when they were symptom-free. Maternal serum levels of the candidate agents PGE2, IL-1beta, and TNFalpha were measured. The study shows a positive relationship between NVP and maternal serum PGE2 levels in early pregnancy. PMID- 11014945 TI - Gamma-delta T cells in midgestation human placental villi. AB - One question that remains is how the immune system at the maternal-fetal interface supports tolerance of the fetus while at the same time protecting it from infection. A potential answer is that local innate immunity is augmented while adaptive immunity is downregulated. In this study, we focus on T cells of the gamma-delta lineage, thought to be important in certain innate responses. Using tissue from normal pregnancies, we documented the presence of gamma-delta T cells and their counterpart, alpha-beta T cells, in midgestation human placental villi. The variable presence of these two T cell lineages in this anatomic site may suggest differential regulation, and herein we describe potential mechanisms for this phenomenon. PMID- 11014946 TI - Use of prostaglandin E2 for cervical ripening in pregnancies with oligohydramnios. AB - The effectiveness of intracervical prostaglandin E2 (PGE2) was studied in 96 primigravidas with unfavorable cervix and need for induction of labor. Group A consisted of 49 oligohydramnios and group B of 47 cases with normal amniotic fluid volume (controls). A single dose of 0.5 mg PGE2 gel was applied intracervically and several parameters were recorded during the next hours. The mean number of uterine contractions increased during the first 2 h in both groups, decreased during the next one and did not change significantly afterwards. The mean fetal heart rate (FHR) in group A decreased during the first 2 h and then increased, staying always within normal limits. The mean FHR in group B increased for 2 h, decreased during the next hour and did not change significantly afterwards. No significant differences were found between the two groups regarding mode of delivery, Apgar score and neonatal acidosis. Intracervical PGE2 appeared to effectively stimulate cervical ripening and labor induction in oligohydramnios, without causing side effects to the uterus and fetus. PMID- 11014947 TI - Perinatal outcome in 41 sets of triplets. AB - OBJECTIVE: To determine the perinatal outcome associated with triplet pregnancies and to compare abdominal delivery with vaginal delivery. METHODS: Retrospective analysis of maternal and neonatal medical records of 41 triplets. 21 were delivered vaginally and 20 were delivered by cesarean section. MAIN OUTCOME: To measure perinatal mortality and early neonatal complications. RESULTS: Between January 1, 1994, and June 30, 1999, there were 41 triplets delivered at our institution. Of these 21 triplets were delivered vaginally and 20 triplets were delivered abdominally. The perinatal mortality rate was 32/123 (26.0%), primarily due to the respiratory distress syndrome. The perinatal deaths are mainly at a birth weight of 500-1,500 g (29/32; 90.6%). Breech presentation was associated with a significantly higher perinatal mortality rate than vertex presentation (62.5 vs. 37.5%). Cesarean delivery was associated with a higher perinatal mortality rate than vaginal delivery (30.0 vs. 22. 2%). CONCLUSIONS: Abdominal delivery in triplets is not superior to vaginal delivery in terms of fetal and early neonatal outcome. The perinatal deaths are increased with low birth weight (500-1,500 g) and with breech presentation. The main cause of neonatal mortality is the respiratory distress syndrome. PMID- 11014948 TI - A case of XY pure gonadal dysgenesis with 46,XYp-/47,XXYp- karyotype whose gonadoblastoma was removed laparoscopically. AB - A case of pure gonadal dysgenesis was investigated. The patient was an 18-year old Japanese woman with a history of primary amenorrhea. She had poorly developed breasts, a hypoplastic uterus, a normal vagina and infantile genitalia. The patient's karyotype was 46,XYp-/ 47,XXYp-. Microsatellite analysis revealed that the X chromosomes of this patient originated from one of the two maternal X chromosomes. DNA analysis of the Y chromosome revealed that she had a deletion of SRY (the sex-determining region on the Y chromosome). She underwent laparoscopic gonadectomies with a final pathology consistent with gonadoblastoma. Laparoscopic surgery is recommended as it is much less invasive and associated with rapid postoperative recovery. PMID- 11014949 TI - Effects of a vasopressin antagonist in women with dysmenorrhea. AB - We compared menstrual pain, uterine contractility and blood circulation, and plasma concentrations of vasopressin and prostaglandin F(2alpha) metabolite in women with versus without primary dysmenorrhea, and determined the effects of a vasopressin antagonist, 1-deamino-2-D-Tyr(OEt)-4-Thr-8-Orn-oxytocin (Atosiban), on these parameters. Our results do not support the contention that vasopressin is involved in the etiology of dysmenorrhea, plasma concentrations of vasopressin being similar in dysmenorrheic women and controls, and the vasopressin antagonist Atosiban having no effect on menstrual pain, intrauterine pressure or uterine artery pulsatility index in dysmenorrheic women. PMID- 11014950 TI - A clinicohysteroscopic scoring system of intrauterine adhesions. AB - We propose a new scoring system for intrauterine adhesions (IUA) that may predict the prognosis after hysteroscopic adhesiolysis. We analyzed hysteroscopic findings and clinical data of patients diagnosed as having IUA by reviewing previously published hysteroscopic classifications of IUA. The data were subjected to evaluation according to the most popular classifications in addition to the proposed classification using a scoring system for different parameters. The results obtained by the proposed scoring system matched well with other classifications regarding grade I and III IUA. But in the cases with moderate IUA (grade II), there was overlap between the classifications (sensitivity 58.3%) which can be attributed to considering the menstrual and reproductive history. Further studies are needed to assess its prognostic value. PMID- 11014951 TI - Use of intramuscular progesterone versus intravenous albumin for the prevention of ovarian hyperstimulation syndrome. AB - This study was designed to compare the effectiveness of intramuscular progesterone with that of intravenous albumin in the prevention of ovarian hyperstimulation syndrome (OHSS). Ninety-six patients at high risk to develop OHSS (estradiol concentration >9, 000 pmol/l on the day of hCG administration and over 20 follicles of a diameter larger than 14 mm observed by transvaginal ultrasonography) and undergoing in vitro fertilization-embryo transfer were enrolled. They were randomly treated with intramuscular progesterone (200 mg/day) or 100 ml of 20% intravenous albumin in order to estimate the difference in the incidence of OHSS. A significant difference in the incidence of moderate OHSS and no cases of severe OHSS were observed between the groups. Our data show the effectiveness in preventing OHSS with high doses of progesterone. PMID- 11014952 TI - Coasting may reduce the severity of the ovarian hyperstimulation syndrome in patients with polycystic ovary syndrome. AB - The aim of our study was to determine the efficacy of postponing administration of human chorionic gonadotropin while continuing daily gonadotropin-releasing hormone agonist therapy ('coasting') to prevent the occurrence of severe ovarian hyperstimulation syndrome (OHSS) for patients with polycystic ovary (PCO) syndrome. Five patients with PCO who had been hospitalized due to severe OHSS in previous in vitro fertilization and embryo transfer or intrauterine insemination cycles at the Tottori University Hospital were included in the study. The rates of mature oocytes and fertilization were comparable between the cycles. A singleton pregnancy was achieved in a patient during the coasting cycle, and none of the women developed severe OHSS in coasting cycles. The results suggest that coasting may be an alternative method for reducing the severity of OHSS in patients with PCO. PMID- 11014953 TI - Effects of hormone replacement therapy on selected indices of immune function in postmenopausal women. AB - The purpose of this study was to examine the effects of long-term hormone replacement therapy (HRT) on selected indices of resting immune function in postmenopausal women. Postmenopausal women aged 54-66 were divided into two groups, those taking HRT (n = 17) and controls (n = 19). Blood samples were obtained and analyzed for mononuclear cell numbers, lymphocyte proliferation (LP) and natural cell-mediated cytotoxicity (NCMC). There were no significant differences between groups for mononuclear cell numbers. LP was significantly higher for HRT, while NCMC was significantly lower for HRT. HRT is currently being prescribed to postmenopausal women for prevention of a variety of medical conditions including osteoporosis, cardiovascular disease, stroke, and Alzheimer's disease yet HRT is often associated with altered immune parameters. In this study, women taking HRT had increased lymphocyte blastogenesis and decreased NCMC compared to controls. PMID- 11014954 TI - Pure versus complicated vulvar vestibulitis: a randomized trial of fluconazole treatment. AB - OBJECTIVE: To examine the effectiveness of a 6-month treatment consisting of a weekly oral dose of 150 mg fluconazole for women with vestibulitis, and to explore the causes of treatment failure. METHODS: Forty women with vestibulitis were randomized to either of two treatment groups. One group received a 6-month low oxalate diet with calcium citrate complement, as a placebo, and the second group the same diet and calcium citrate with the addition of a weekly oral tablet of 150 mg fluconazole. The women were examined 3 months after completing treatment, for response to therapy. RESULTS: The addition of intensive 6-month fluconazole treatment did not lead to an outcome better than that attained by maintaining a low oxalate diet with calcium citrate supplementation. The satisfactory response rate was 15 and 30%, respectively. The presence of 'complicated vestibulitis', candidiasis concomitant with vestibulitis, decreases the satisfactory response rate regardless of the type of treatment administered (odds ratio 19.9, 95% CI 1.6, 250). CONCLUSION: Prolonged oral fluconazole is an ineffective treatment of vestibulitis, whether pure or complicated by concomitant vulvovaginal candidiasis. The coexistence of candidiasis and vestibulitis - complicated vestibulitis - might represent a subset of vestibulitis that is resistant to the currently available medical therapy. PMID- 11014955 TI - Expression of constitutively active c-MET receptor in human choriocarcinoma. AB - OBJECTIVE: The c-MET oncogene encodes the transmembrane tyrosine kinase receptor for hepatocyte growth factor (HGF). Recently, either HGF or c-MET has been knocked out in the mouse, and the mutation is associated with defects in the development of the placenta by means of the reduction of trophoblast cells. We examined the role of c-MET in malignant transformation of trophoblast cells. METHODS: In four human choriocarcinoma cell lines, GCH-1, GCH-1m, NUC-1 and SCH, the expression of HGF and c-MET was analyzed by RT-PCR and Western blotting using anti-c-MET antibody and antiphosphotyrosine antibody. RESULTS: RT-PCR analysis showed that no cell lines expressed HGF, however all the cell lines revealed the expression of c-MET mRNA. c-MET receptor was expressed and also tyrosine phosphorylated constitutively in these cell lines by Western blotting. CONCLUSION: These results suggest that c-MET may be involved in the growth and behavior of human choriocarcinoma although an autocrine fashion of HGF is unlikely. PMID- 11014956 TI - Primary peritoneal carcinoma: a report of twelve cases and a review of the literature. AB - Primary peritoneal carcinoma (PPC) is rare tumor histologically identical to epithelial ovarian carcinoma (EOC); it is differentiated from EOC based on the extent of gross ovarian involvement and microscopic invasion of the cortex. We report 12 cases of PPC which were diagnosed in our Department during a 9-year period. Total abdominal hysterectomy, bilateral salpingo-oophorectomy and omentectomy were performed in 9 patients, while 3 underwent only explorative laparotomy with bilateral salpingo-oophorectomy. All patients were treated with postoperative platinum-based chemotherapy. After a median follow-up of 42 months, only 5 patients are alive without disease. PPC is a rare tumor currently managed in the same way as ovarian cancer. Primary debulking surgery and chemotherapy represent the cornerstones of treatment. Considering the limited number of patients with PPC, no definitive conclusion can be drawn concerning the prognostic factors for survival. PMID- 11014957 TI - Analysis of TH1 and TH2 cells by intracellular cytokine detection with flow cytometry in patients with ovarian cancer. AB - OBJECTIVE: Our aim is to assess the immune status of patients with ovarian cancer by analyzing the ratio of T helper type 1 (TH1) to T helper type 2 (TH2) populations in peripheral blood lymphocytes (PBL). METHODS: We examined TH1/TH2 ratios in PBL obtained from 21 ovarian cancer patients who had just received postoperative chemotherapy, by detecting the intracellular IFN-gamma and IL-4 production with 3-color flow cytometry. Additionally, we evaluated the influence of a granulocyte-colony stimulating factor (G-CSF) injection on TH1 and TH2 populations for a rescue of granulocytopenia due to the chemotherapy. RESULTS: We could not find any significant difference of the TH1/TH2 ratios in terms of age, International Federation of Gynecology and Obstetrics (FIGO) clinical stage and clinical tumor status. As for the clinical tumor status, however, the patients with residual cancer had a higher TH1/TH2 ratio, though it was not statistically significant (p = 0. 15). Anticancer chemotherapy is also considered to lead to the immunosuppressive state of the patients. TH1 and TH2 populations of PBL in the patients during chemotherapy showed an unfavorable imbalance that was shifted from TH1 to TH2 10 days after anticancer drug administration (p = 0.049). G-CSF administration, on the other hand, was likely to induce a cell population shift from TH2 to TH1 assessed by the intracellular cytokine assay (p = 0.051), and never induced an unfavorable imbalance from TH1 to TH2 in the T cell population by a 1-day injection of G-CSF. CONCLUSION: Together, these data indicate that the TH1/TH2 ratio analyzed by intracellular cytokine flow cytometry seems to be a good indicator to assess the immune status in cancer. PMID- 11014958 TI - Uterine rupture associated with vaginal birth after cesarean section: a complication of intravaginal misoprostol? AB - Intravaginal misoprostol has become increasingly employed for labor induction among patients with an unfavorable Bishop's score. Almost all of the reported studies have specifically excluded patients with prior uterine surgery. There has been, therefore, very little information concerning its usage among patients attempting vaginal birth after cesarean section. We report a patient with two prior low transverse uterine incisions who experienced uterine rupture after having received a single 25-microg intravaginal dose of misoprostol. PMID- 11014959 TI - Human papillomavirus genotypes in cervical cancer with different histological features. AB - We report the case of a 32-year-old woman having developed two cervical cancers synchronously, an adenocarcinoma and a squamous cell carcinoma. Polymerase chain reaction with the general primers GP5/GP6 and a subsequent enzyme-linked immunosorbent assay to detect human papillomaviruses (HPV) resulted in isolation of HPV 33 in the squamous cell carcinoma and HPV 18 in the adenocarcinoma. This is the first reported case of two histologically different synchronous cervical cancers with this distinct HPV expression pattern, and further confirms the association of certain 'high-risk' HPV genotypes to different histological features of carcinoma. Furthermore, the important role of microdissection for gaining tumor tissue of different areas in molecular diagnostics is supported. PMID- 11014960 TI - Paroxysmal nocturnal hemoglobinuria and the risk of venous thrombosis: review and recommendations for management of the pregnant and nonpregnant patient. AB - BACKGROUND: Paroxysmal nocturnal hemoglobinuria is a rare, clonal primitive hematopoietic cell disorder, often affecting middle-aged adults, including women of reproductive age. Major morbidity and mortality with this disease are often ascribed to the development of venous thromboembolism. We reviewed the current literature on the risk of venous thrombosis among nonpregnant and pregnant patients, and generated recommendations for the prevention of venous thromboembolism, as well as duration of treatment for affected patients who develop thrombotic disease. METHODS: We searched Medline for papers published between January 1966 and April 1999. We also requested relevant unpublished data from speakers who attended a recent international workshop of paroxysmal nocturnal hemoglobinuria. References from all primary data and review publications were also examined. Only English language publications were included. Event rates for venous thromboembolism and death were pooled using a random effect technique. Reports of paroxysmal nocturnal hemoglobinuria during pregnancy were summarized using descriptive statistics only. RESULTS: Thirteen retrospective studies of paroxysmal nocturnal hemoglobinuria in nonpregnant individuals were found. The rates of venous thrombosis varied considerably, but were reported to affect 14.4% of all individuals [95% confidence interval (CI) 7.6-25.5]. Among patients from western nations, venous thromboembolism seemed to develop at a higher rate (30.3%, 95% CI 26. 1-34.9). The majority of venous thromboembolic events were intra-abdominal, principally within the hepatic and mesenteric veins. The likely cause of death among patients with paroxysmal nocturnal hemoglobinuria was described in nine studies: 22.2% of fatalities were due to venous thrombosis (95% CI 11.8-38.0), more commonly in western countries (event rate 37.2%, 95% CI 21.6-56.0). Another 20 published reports described the outcome of 33 pregnant women with paroxysmal nocturnal hemoglobinuria. Two women developed venous thromboembolism during pregnancy and another 2 during the postpartum state for a combined event rate of 12.1% (95% CI 3.4-25.2), three of which resulted in death. The all-cause mortality rate was 20.8% (95% CI 7.3 39.0). Both anemia (event rate 72.7%, 95% CI 56.5-86.3), and thrombocytopenia (event rate 27.3%, 95% CI 13.7-43.5) were common, often necessitating red cell or platelet transfusions. Almost half of all infants (54.8%, 95% CI 36.1-72.7) were delivered preterm, and had a mean live birth weight of 2,800 g. Three of 34 reported births ended in death (perinatal mortality rate 8.8%, 95% C 1.9-23.7). CONCLUSION: In accordance with the apparently high rate of venous thrombosis among pregnant and nonpregnant individuals with paroxysmal nocturnal hemoglobinuria, especially for fatal thrombosis, we developed practical recommendations for the prevention and treatment of venous thromboembolic disease in these groups. PMID- 11014961 TI - Factor V (His 1299 Arg) in young Turkish patients with cerebral infarct. AB - Inherited gene defects related to the coagulation system have been reported as risk factors for stroke. Recently, a genetic component in the factor V (FV) gene that contributes to activated protein C resistance both in the presence and absence of FV 1691 G-->A was reported. This highly conserved FV gene haplotype was marked as R2 polymorphism, an A to G alteration at position 4070 in exon 13 that predicts the His 1299 Arg substitutions. The aim of this study was to evaluate the role of this mutation in Turkish children with ischemic infarct. The case-control study included 48 patients with cerebral infarction; all were 18 years of age or younger (range: 10 months to 18 years). Ten (20.8%) of the 48 patients were found to carry the FV 1299 His-->Arg mutation, one being homozygous. One patient who had a combination of FV 1691 G-->A and protein C deficiency also carried the FV 4070A mutation. A homozygous FV 1299A patient had a prothrombin (PT) 20210A mutation in the heterozygous state. The cerebral infarct risk for FV 1299 was found to be 2.4 (95% confidence interval 0.9-6.8) for all groups. When underlying conditions were excluded, the incidence of FV 1299 was found to be 8/35 (22.8%), but the risk was almost the same. When two other common thrombophilic mutations (i.e. FV 1691 G-->A and PT 20210 G-->A) were excluded, the incidence of FV 4070 mutation increased to 7/21 (33.3%). The risk also increased to 3.9 (95% confidence interval 1.2-12.3). PMID- 11014962 TI - Influence of fibrinogen degradation products on thrombin time, activated partial thromboplastin time and prothrombin time of canine plasma. AB - To investigate how thrombin time, activated partial thromboplastin time (APTT) and prothrombin time are influenced by fibrinogen degradation products (FDP), different concentrations (0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8 and 1.0 mg/ml) of the purified FDP X, Y, D and E were added to the plasma of healthy dogs. If fragment Y was added to the plasma a considerable inhibitory effect could be demonstrated for all three test systems. A significant prolongation (p < 0.05) was found for concentrations of > or =0.1 mg/ml (thrombin time, APTT) and > or =0.2 mg/ml (prothrombin time). With FDP Y concentrations from >0.185 mg/ml (prothrombin time) to >0.24 mg/ml (APTT) coagulation time was prolonged beyond the respective reference range. As regards the other fragments, a comparable inhibitory effect could only be shown for fragment X added to the thrombin time test system. This effect can most probably be explained by the competition of the FDP X and fibrinogen for the fibrinogen binding sites of thrombin, rather than by a fibrin polymerization disorder. The results demonstrate that for plasma with normal fibrinogen concentration the group tests are only prolonged beyond the reference range at FDP concentrations very rarely found in spontaneous hyperfibrinolysis. PMID- 11014963 TI - Comparison of different methods to measure fibrinogen concentration in canine plasma with respect to their sensitivity towards the fibrinogen degradation products X, Y and D. AB - In this study, fibrinogen measurements according to the Clauss method, photometric method and Jacobsson method have been investigated to find out how they are influenced by adding in vitro the purified canine fibrinogen degradation products (FDP) X, Y and D. Test results according to the Clauss method were found to be underestimated if the fragments X, Y and D were added while measurements according to the Jacobsson method turned out to underestimate the real fibrinogen concentration if the FDP Y and D were added. The Clauss method was particularly sensitive towards FDP. Results were considerably underestimated even with a quantity as little as 0.05 g FDP Y or FDP D/g fibrinogen (p < 0.05). The photometric method was only affected by FDP X leading to false high results. If FDP X was added, fibrinogen values were also overestimated with the Jacobsson method. Our results demonstrate that the photometric method is the most accurate. PMID- 11014965 TI - Endothelial function, variables of fibrinolysis and coagulation in smokers and healthy controls. AB - OBJECTIVE: To asses endothelial function and variables of fibrinolysis and coagulation in smokers compared to healthy controls. METHODS: Flow-associated dilation as a marker for peripheral endothelial function and intima media thickness as a marker for early morphologic vascular changes were measured in otherwise healthy smokers (n = 30, 16 males and 14 females, age: 40.6 +/- 11.3 years, body mass index 24.9 +/- 3.7 kg/m(2)) and non-smoking controls matched for age and sex using high-resolution ultrasound. Variables of the coagulation system (thrombin-antithrombin III complex, fibrinogen) and fibrinolysis (tissue plasminogen activator, plasmin-alpha(2)-antiplasmin complex) were determined by ELISA and plasminogen activator inhibitor activity by means of a chromogenic substrate test. RESULTS: Compared to the non-smoking controls, flow-associated vasodilatation was significantly reduced (6.9 +/- 4. 4 vs. 10.5 +/- 6.2%, p = 0.01) and intima media thickness tended to be increased (0.58 +/- 0.12 vs. 0.52 +/- 0.14 mm, p = 0.08) in smokers. The thrombin-antithrombin III complex, fibrinogen, plasmin-alpha(2)-antiplasmin complex, tissue-plasminogen activator and plasminogen activator inhibitor activity did not differ between smokers and controls. CONCLUSION: Our data indicate that peripheral endothelial dysfunction is common in smokers even without major alterations in molecular markers of the coagulation and fibrinolysis system. PMID- 11014964 TI - Characterization of murine anti-glycoprotein Ib monoclonal antibodies that differentiate between shear-induced and ristocetin/botrocetin-induced glycoprotein Ib-von Willebrand factor interaction. AB - Platelet adhesion to vascular subendothelium under conditions of high shear stress is mediated by the platelet glycoprotein (GP) Ib-von Willebrand Factor (vWF) interaction. The aim of this study was to characterize the murine monoclonal antibodies (MoAbs) 27A10 and 28E6, both raised against purified GPIb. The MoAb 27A10 is a potent inhibitor of shear-induced platelet adhesion to collagen type I in a flow chamber at shear rates of 1,300 and 2,700 s(-1). 20 microg/ml of MoAb 27A10, furthermore, could completely block shear-induced aggregation in a modified Couette viscometer at shear rates of 1,000 and 4,000 s( 1). On the other hand, MoAb 27A10 had a negligible effect on botrocetin-induced GPIb-vWF binding and is only a poor inhibitor of the ristocetin-dependent interaction. In contrast, MoAb 28E6 did abolish both the ristocetin- and botrocetin-induced GPIb-vWF binding, whereas it did not block the shear-induced interaction. Thus, we identify here two anti-GPIb MoAbs 27A10 and 28E6 that either preferentially inhibit the shear-induced or the ristocetin/botrocetin induced platelet-vWF interaction. With these tools it should be possible to more clearly define the mechanisms by which platelets bind to vWF in vivo. PMID- 11014966 TI - Antinuclear antibody- and extractable nuclear antigen-related diseases. AB - In 1948, the observation of the LE cell phenomenon in a patient with systemic lupus erythematosus (SLE) began the discovery of a broad variety of autoantibodies directed to nuclear antigens called antinuclear antibodies (ANA). Nowadays, different ANA serve as important diagnostic parameters for differentiating most of the connective tissue diseases, such as SLE, neonatal lupus syndromes, Sjogren's syndrome, scleroderma, autoimmune myositis, mixed connective tissue disease and other overlaps. This overview summarizes the history of ANA and their detection methods, in part to introduce the subsequent papers dealing with special topics of ANA-related diseases in this issue. Furthermore, the pathogenic role of these autoantibodies in targeting non-organ specific intracellular antigens as a functional important constituent of a subcellular particle or multimolecular complex is addressed. Notably, some of these autoantibodies have functioned as significant tools for cell biologists to elucidate the subcellular structures and functions of these autoantigens. In the future, we can expect further advances to answer such important questions as why these antigens are targets of autoantibodies, what is their pathogenic impact and what are the triggers of autoimmunity? PMID- 11014967 TI - The idiotypic network in antinuclear-antibody-associated diseases. AB - Antinuclear antibodies (ANA) entail a large group of autoantibodies (Abs) that bind certain nuclear antigens. The ANA test is a useful screening test for many autoimmune diseases and the presence of a specific binding pattern directs secondary testing for specific Abs associated with the suspected disease. Idiotypes (Ids) are the antigenic constitution of the variable region of an Ab, and they are recognized by anti-Ids Abs. The Id network is composed of interacting Abs in which the Id determinants of each Ab are complemented by those of another. It has a role in both physiologic and pathologic conditions. In this communication, we review the induction of autoimmune diseases via Id manipulation, the Ids of some ANA-associated Abs (DNA, SS-A, SS- B, Sm Abs), the pathogenic role of Abs carrying Ids, and the clinical implications of the Id network in autoimmunity. PMID- 11014968 TI - Antinuclear autoantibodies: fluorescent highlights on structure and function in the nucleus. AB - The eukaryotic nucleus is dynamically organized with respect to particular activities, such as RNA transcription, RNA processing or DNA replication. The spatial separation of metabolic activities is best reflected by the identification of functionally related proteins, in particular substructures of the nucleus. In a variety of human diseases, the integrity of such structures can be compromised, thus underlining the importance of a proper nuclear architecture for cell viability. Besides their clinical relevance, antinuclear autoantibodies (ANAs) have contributed to a large extent to the identification of subnuclear compartments, the isolation and cloning of their components (the autoantigens), as well a the characterization of their function. Although sophisticated techniques, such as confocal laser scanning microscopy (CLSM), fluorescence resonance energy transfer (FRET) and in vivo observation of cellular events have recently been established as valuable tools to study subnuclear architecture and function, cell biologists will continue to appreciate the specificity and power of ANAs for their research. PMID- 11014969 TI - Etiopathogenesis of systemic lupus erythematosus. AB - Systemic lupus erythematosus is an autoimmune disease of unknown etiology. Research efforts of the last few years have mainly focused on basic molecular and cellular pathogenetic processes of the disease. Consequently, this paper reviews the etiopathogenetic hallmarks, such as impaired amount and presentation of nuclear antigens, production of antinuclear antibodies by T-cell-dependent B cell stimulation and organ damage by anti-dsDNA antibodies or immune complexes that are discussed at the present time. In summary, the hypothesis of a dysregulation of apoptotic cell clearance is strongly supported and broadly discussed. PMID- 11014970 TI - Use of immunoglobulin variable-region genes by normal subjects and patients with systemic lupus erythematosus. AB - Antibodies to specific autoantigens are serological hallmarks of systemic autoimmune diseases. These autoantibodies are thought to represent a consequence of immune dysregulation in these conditions, and, in part, have been shown to be involved in their pathologic consequences. However, the mechanisms that lead to the production of autoantibodies are still unknown. The observation that certain autoantibodies are frequently encoded by a limited number of immunoglobulin (Ig) variable-region gene segments suggested that a bias in the development of the Ig repertoire might play a role in the tendency to develop autoimmunity. Whether the use of these individual gene segments is random or different in normal subjects and patients with systemic autoimmune disorders remains a matter of controversy. New approaches for the analysis of variable-region genes from unstimulated individual human B cells employing the single-cell polymerase chain reaction have provided new insights in the B cell repertoire of both normal subjects and patients with systemic autoimmune diseases. Using this approach, the analysis of nonproductive and productive Ig variable-region gene rearrangements made it possible to distinguish molecular processes, as manifested in the nonproductive repertoire, from subsequent selection influences. An initial study in a patient with systemic lupus erythematosus has led to the hypothesis that the molecular generation of the B cell repertoire is similar in patients and normal subjects but subsequent influences and, most notably, extensive mutations and receptor editing differ significantly in shaping the peripheral IgV gene use by persons with autoimmune diseases. PMID- 11014971 TI - Sjogren's syndrome: autoantibodies to cellular antigens. Clinical and molecular aspects. AB - Autoantibodies to cellular autoantigens are usually found in sera of patients with systemic autoimmune rheumatic diseases. Patients with Sjogren's syndrome (SS) frequently present autoantibodies to both organ and non-organ-specific autoantigens. The most commonly detected autoantibodies are those directed against the ribonucleoproteins Ro/SSA and La/SSB. The presence of the antibodies in SS is associated with early disease onset, longer disease duration, parotid gland enlargement, higher frequency of extraglandular manifestations and more intense lymphocytic infiltration of the minor salivary glands. Over the past several years, the structure and function of these autoantigens have been extensively studied. Several centers, using different techniques, have investigated the B cell epitopes on the protein components Ro 60 kD, Ro 52kD, and La 48 kD. Finally, increased evidence of direct involvement of anti-Ro/SSA and anti-La/SSB autoantibodies in the pathogenesis of tissue injury has been contributed by several studies. PMID- 11014972 TI - Significance of autoantibodies in neonatal lupus erythematosus. AB - Autoantibodies produced by the mother and transported into the fetal circulation are of significant importance in the diagnosis of neonatal lupus syndromes. These humoral autoimmune findings provide an unique opportunity to assess the pathogenic role of autoantibodies against the Ro(SS-A)/La(SS-B) complex, most notably for congenital heart block. Current knowledge about the involved autoantibody-autoantigen systems, including recent therapeutic concepts of these autoimmune syndromes, is summarized. PMID- 11014973 TI - Antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is a disease characterized by venous and arterial thromboses or spontaneous abortions and the repeated detection of antiphospholipid antibodies (aPL). APS may be associated with another autoimmune disease (secondary APS), particularly systemic lupus erythematosus (SLE), or unrelated to an underlying disease (primary APS). APS affects almost all organs. In addition to the clinical criteria, lupus anticoagulant testing and immunological aPL determinations are required to establish the diagnosis of APS. PMID- 11014974 TI - Diagnostic and prognostic relevance of autoantibodies in uranium miners. AB - Uranium miners exposed to silica dust have a higher risk of developing systemic sclerosis (SSc) and systemic lupus erythematosus (SLE). Sera of 1976 former uranium miners were analysed for autoantibodies typical of connective tissue disease. The frequency of some of these antibodies (anti-centromere, topoisomerase I, -nucleolar, -dsDNA, -Ro/SSA, -La-SSB and U1-RNP antibodies) was significantly higher compared to a gender- and age-matched control group and was associated with the intensity of exposure as well as with clinical symptoms of SSc or SLE. It was also shown that SSc-associated autoantibodies may serve as an early indicator of disease development. Some differences in the autoantibody production between silica-dust-associated and idiopathic SLE/SSc were observed that might be caused by environmental factors in the population of uranium miners. PMID- 11014975 TI - Diagnostic importance of anti-proteasome antibodies. AB - 20S proteasome represents the proteolytic core complex for cytoplasmic protein degradation that is involved in the activation and regulation of the immune response. In this context, proteasome generates antigenic peptides for the MHC class I pathway and activates NF-kappaB. In a recent analysis, we could identify a frequent humoral autoimmune response directed against specific proteasomal subunits in patients with autoimmune myositis, systemic lupus erythematosus and primary Sjogren's syndrome. The outer ring subunit HC9(alpha3) was identified as the predominant target of the anti-proteasome response in these entities. In addition to the reactivity against HC9(alpha3), patients with primary Sjogren's syndrome expressed a more polyspecific recognition pattern of proteasomal subunits involving the active inner ring proteins. In follow-up analysis, anti proteasome antibody titers revealed a correlation with disease activity in patients with autoimmune myositis and systemic lupus erythematosus. The current review summarizes recent data providing evidence that the 20S proteasome represents an important target of the humoral autoimmune response in systemic autoimmune diseases and extends insight into pathogenic aspects of these diseases. PMID- 11014976 TI - Role of apoptosis in pathogenesis and progression of renal diseases. PMID- 11014977 TI - The obsession with high dietary protein intake in ESRD patients on dialysis: is it justified? AB - BACKGROUND: The belief that dialysis patients require a much higher intake of protein than the average person is widely held among nephrologists and a recent editorial review on the care of patients undergoing hemodialysis advised a protein intake of at least 1.5 g/kg/day in this population. In theory, the requirements for daily protein intake of dialysis patients should be the same as normals plus whatever extra obligatory nitrogen losses are imposed by the dialytic process. Since the average loss of protein and/or amino acids is about 8 g/day in peritoneal dialysis and 8 g per session in hemodialysis patients, the daily intake of protein in dialysis patients needs to be increased only by this small magnitude. METHODS: All the literature reporting on nitrogen balance data on different protein intakes in chronic maintenance dialysis patients was reviewed and analyzed. RESULTS: There were 3 balance studies addressing this issue in hemodialysis patients and 6 in peritoneal dialysis patients. These data show that dialysis patients are in nitrogen balance even with a protein intake as low as 0.7 g/kg/day. CONCLUSIONS: The nitrogen balance data confirm that in stable dialysis patients a safe requirement of protein intake is about 1 g/kg/day and attempts at increasing protein intake beyond this value are not warranted. The persistent recommendation of a high dietary intake of protein for dialysis patients may reflect the influence of two other issues: the belief that dialysis is a catabolic event, and the data showing an inverse correlation between protein intake and outcome. PMID- 11014978 TI - Urinary retinol-binding protein as a prognostic marker in the treatment of nephrotic syndrome. AB - We studied the urinary levels of retinol-binding protein (urRBP), an index of proximal tubular dysfunction, in patients with nephrotic syndrome before and approximately 2 months after the beginning of steroid therapy as a predictor of response to therapy which included for some patients courses of immunosuppressive drugs. Those patients with minimal-change disease, mesangial proliferative glomerulonephritis, and focal-segmental glomerulosclerosis who had normal pretreatment urRBP levels were responsive to treatment; occasionally, responsive patients had an initially elevated urRBP level which normalized during treatment. Contrariwise, those patients with abnormally high levels of urRBP which did not normalize during treatment did not respond to treatment. The chance of a patient with minimal-change disease, mesangial proliferative glomerulonephritis, or focal segmental glomerulosclerosis and a pretreatment urRBP level equal to or >1.0 mg/l being resistant to steroid treatment is 30 times that of a patient with a urRBP level <1.0 mg/l and even higher, if we consider the levels obtained during treatment. PMID- 11014979 TI - Angiotensin-converting enzyme insertion/deletion polymorphism and prognosis of IgA nephropathy. AB - BACKGROUND/AIM: Well-known factors for a poor prognosis in IgA nephropathy (IgAN) are hypertension, proteinuria, and renal insufficiency at the time of diagnosis. Also hypertriglyceridemia and hyperuricemia seem to play a role in the progression of IgAN. Angiotensin-converting enzyme (ACE) gene I/D polymorphism has been associated with cardiovascular diseases and with progression of IgAN. We, therefore, investigated the contribution of ACE gene I/D polymorphism in the prognosis of IgAN and its association with the other risk factors affecting the prognosis. METHODS: A total of 168 patients with IgAN were followed up for 6-17 (median 11) years from renal biopsy with respect to progression of renal disease defined as elevation of serum creatinine above 125 microM (1.4 mg/dl) in men or 105 microM (1.2 mg/dl) in women and over 20% from the baseline level. In addition to serum creatinine, the urinary protein excretion was evaluated at the time of renal biopsy and at the assessment visit at the end of the follow-up period. RESULTS: During the follow-up period, 26 (15%) patients showed progression of renal disease. Patients with ACE genotype II had a more favorable course than those with genotypes ID or DD. Although there were no significant differences among the ACE genotypes with respect to proteinuria > or =1 g/24 h at the time of renal biopsy, proteinuria > or =1 g/24 h was more frequent in patients with genotypes ID or DD than in those with genotype II at the end of the follow-up period. No associations were found between hypertension, serum lipids or serum urate, and ACE genotypes. CONCLUSIONS: Our results show that patients with ACE genotype II have a more favorable prognosis than those with genotypes ID/DD. Secondly, proteinuria (> or =1 g/24 h) found in patients with genotype II at diagnosis may improve, while in patients with genotypes ID/DD it is a more constant feature. PMID- 11014980 TI - Oral supplementation with gamma-linolenic acid extracted from Mucor circinelloides improves the deformability of red blood cells in hemodialysis patients. AB - BACKGROUND: The development of abnormalities in red blood cell (RBC) deformability in patients undergoing hemodialysis remains a major problem, because it is related to peripheral microcirculation, oxygen supply, and various complications in such patients. gamma-Linolenic acid (GLA; 18:3n-6), one of the polyunsaturated fatty acids and a precursor of prostaglandin E(1), is reported to have a favorable effect on the deformability of circulating blood cells in diabetic patients. METHODS: In order to clarify the efficacy of GLA on RBC deformability in 7 patients undergoing maintenance hemodialysis, we examined in a pilot study the changes in the deformability of RBC and the changes in the phospholipid fatty acid composition in both plasma and RBC membrane before and after high-dose oral supplementation with GLA derived from Mucor circinelloides for 12 weeks. RESULTS: Before supplementation, the micropore passage time of RBC suspension, which is an indicator of RBC deformability, in these patients was markedly longer than that in healthy control subjects. After administering GLA, the prolonged passage time of the patients both rapidly and steadily decreased and nearly reached control levels. Light microscopic observations of RBCs using Giemsa stain revealed a decreased number of poikilocytes after supplementation. An analysis of the fatty acid composition before treatment and 8 weeks after starting the treatment showed the dihomo-gamma-linolenic acid (DGLA; 20:3n-6) level in the plasma to have increased (p < 0.05), while the arachidonic acid (AA; 20:4n-6) concentration in the RBC membrane decreased (p < 0.05). The level of DGLA in the RBC membrane, the level of GLA, and the ratio of GLA + DGLA/AA in plasma and RBC membrane did not change significantly; however, these all tended to increase. CONCLUSION: The results of this pilot study indicate that the oral supplementation of GLA extracted from M. circinelloides improves the poor RBC deformability in hemodialysis patients, partly by inducing changes in the composition of fatty acids in plasma and RBC membrane. PMID- 11014982 TI - Moderate metabolic acidosis and its effects on serum parameters in hemodialysis patients. AB - We screened the laboratory data of 50 chronic hemodialysis patients selected randomly over a 21-month period to generate 158 data points which identified two groups: (1) those with a predialysis total CO(2) concentration less than or equal to 19 mEq/l (data A; n = 57) and (2) those with a predialysis total CO(2) concentration greater than 19 mEq/l (data B; n = 101). Then, both groups were compared for the following parameters: predialysis blood urea nitrogen (BUN), serum phosphorus, uric acid, creatinine, and albumin concentrations, Kt/V, urea reduction ratio, normalized protein catabolic rate, dry weight, ultrafiltration, blood flow and dialysis flow rates, duration of dialysis treatment, and blood pressure. Group data A had significantly higher predialysis BUN, phosphorus, and uric acid concentrations than group data B. There were significant inverse correlations between predialysis serum bicarbonate and predialysis BUN, phosphorus, and uric acid concentrations. Although it is not clear what the long term side effects of moderate metabolic acidosis are, we recommend its correction. PMID- 11014981 TI - Long-term knowledge retention following predialysis psychoeducational intervention. AB - BACKGROUND/AIMS: Early identification and predialysis psychoeducation are gaining acceptance. Although research supports the immediate value of predialysis interventions, long-term benefits remain unknown. We examined long-term knowledge retention following a psychoeducational intervention. METHODS: 47 progressive renal failure patients completed the Kidney Disease Questionnaire at baseline and 18, 30, 42, and 54 months after initiating renal replacement therapy (RRT; the 'longitudinal' sample). A larger cohort provided data at one or more of these points (n = 132, 117, 101, and 70 at 18, 30, 42, and 54 months, respectively; the 'cross-sectional' sample). RESULTS: Initial knowledge gains among psychoeducation recipients were followed by a significant knowledge advantage for three groups throughout follow-up. Patients who received predialysis psychoeducation either before or after starting dialysis demonstrated superior Kidney Disease Questionnaire scores as compared with those identified before the initiation of RRT who received the usual standard of practice. Patients identified after the initiation of RRT and who received standard education, however, demonstrated the same level of knowledge retention as produced by psychoeducation. The results were identical across the longitudinal and cross-sectional samples. CONCLUSIONS: Patient education produces important benefits in end-stage renal disease, but the incremental value of early intervention remains to be demonstrated. PMID- 11014983 TI - Constant, but not pulsed calcitriol suppresses hemodialysis patients' antigen induced lymphocyte proliferation. AB - BACKGROUND/AIMS: In vitro constant calcitriol [1,25-(OH)(2)D(3)] inhibits healthy individuals' T lymphocyte proliferation at supraphysiological concentrations. In contrast, among hemodialysis patients, intravenous 1,25-(OH)(2)D(3) pulse therapy of secondary hyperparathyroidism has been shown to be even immunostimulatory. We studied the effect of in vitro constant and intermittent 1, 25-(OH)(2)D(3) on lymphocyte antigen response of hemodialysis patients. METHODS: Twelve hemodialysis patients' peripheral blood mononuclear cells were stimulated with purified protein derivative of tuberculin (12.5, 25 and 50 mg/l) or tetanus toxoid (TT; 1,000, 5, 000 and 10,000 Lf/l, limit of flocculation) for 7 days. Constant 1, 25-(OH)(2)D(3) was added to all cultures at concentrations of 0, 10( 10) or 0.25 x 10(-9) mol/l (0, 42 and 105 ng/l) and to half of the cultures additionally as a 0.75 x 10(-9) mmol/l (315-ng/l) pulse on the 5th culture day. RESULTS: TT-induced lymphocyte proliferation was statistically related to a constant 1,25-(OH)(2)D(3) concentration (p = 0.001, analysis of variance). With constant 1, 25-(OH)(2)D(3) concentrations of 0, 42 and 105 ng/l, the TT-induced responses were 1.53, 1.44 and 1.40 log cpm, respectively (mean of TT concentrations). The responses of the (additionally) pulse-treated cells [1.65, 1.50 and 1.40 log cpm; concentrations of constant 1, 25-(OH)(2)D(3) as above] were similar to those of the nonpulsed cells. Thus constant, but not pulsed 1,25 (OH)(2)D(3) decreased the TT responses. On the purified protein derivative of tuberculin response, neither constant nor pulsed 1,25-(OH)(2)D(3) had any significant effect. CONCLUSIONS: The decline of TT response with constant 1,25 (OH)(2)D(3) corresponds with findings on immunosuppressive action of 1,25 (OH)(2)D(3) in previous studies done on normal subjects' cells. This was not seen with intermittently applied 1,25-(OH)(2)D(3). These results support the previous concept that intermittent 1,25(OH)(2)D(3) therapy is not immunosuppressive in hemodialysis patients. PMID- 11014984 TI - Decreased matrix metalloproteinase activity in the kidneys of hereditary nephrotic mice (ICGN strain). AB - Abnormalities of extracellular matrix (ECM) metabolism, i.e., overproduction and/or inhibition of ECM breakdown, may contribute to progression of fibrotic degeneration in the kidney. Earlier studies revealed that major ECM components, type I, III, and IV collagens, etc., were accumulated in glomeruli and tubulointerstitium in kidneys of Institute of Cancer Research (ICR) derived glomerulonephritis (ICGN) mice which are a novel inbred strain of mice with a hereditary nephrotic syndrome of unknown etiology and are considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we compared the activities of matrix metalloproteinases (MMPs), a family of enzymes that degrade ECM components, in the kidneys of aged ICGN mice and age-matched ICR mice as normal controls. We biochemically measured interstitial collagenase (MMP-1), gelatinase (MMP-2 and MMP-9), and stromelysin (MMP-3) activities in the kidney tissues. Lower activities of MMP-1 and MMP-2 and MMP-9 were demonstrated in the kidneys of ICGN mice as compared with those of ICR mice, but there were no significant differences in the MMP-3 activities between these strains. These results show that decreased MMP activities cause abnormal accumulation of ECM in ICGN mouse kidneys. PMID- 11014985 TI - Complement-mediated killing of mesangial cells in experimental glomerulonephritis: cell death by a combination of apoptosis and necrosis. AB - Immune system mediated, particularly antibody- and complement-mediated, glomerular injury triggers glomerulonephritis (GN). To characterize complement mediated cytotoxicity in GN, we assessed the process of mesangial cell death induced by C5b-9 attack in Thy-1 GN. Cell injury was recognized morphologically, and nuclear DNA breaks were confirmed by the DNA nick end labeling (TUNEL) method as well as DNA gel electrophoresis. Thy-1 GN was induced in rats with anti-Thy 1.1 antibody injection. Mouse IgG (administered antibody) and rat C3 were detected in all glomeruli within 5 min after antibody injection. Damaged mesangial cells with condensed as well as TUNEL-positive nuclei could be observed at 20 min and became prominent at 40-60 min. Ultrastructurally, damaged mesangial cells contained condensed apoptotic nuclei from 40 to 60 min, whereas the cytoplasm showed necrotic degeneration. This was followed by progressive lysis of both nuclei and cytoplasm. The DNA 'ladder' pattern was observed by gel electrophoresis of extracted DNA between 40 and 60 min and correlated with the increased number of TUNEL-positive damaged mesangial cells. To examine the role of complement in this form of cell death, complement depletion was induced in rats by cobra venom factor. Complement-depleted rats showed no rat C3 deposition, rare TUNEL-positive mesangial cells, rare ultrastructural degenerated mesangial cells with apoptotic nuclei and necrotic cytoplasm, and no DNA 'ladder' pattern on gel electrophoresis at 40 min, although prominent mouse IgG was seen in glomeruli. To analyze milder forms of complement injury, a low dose of the antibody was administered to rats with a normal complement level. A few TUNEL positive mesangial cells were detected in the glomeruli which contained apoptotic nuclei and necrotic cytoplasm. Our results indicate that an apoptotic death mechanism accompanies cell necrosis in complement-mediated mesangial cell destruction in GN and that this unusual form of cell death may represent a combination of apoptosis-necrosis within the same cell. Complement injury activates a 'death program' which in turn leads to irreversible damage of mesangial cells and which may contribute to initiation and development of GN. PMID- 11014986 TI - Effects of chronic hypoxia on renal PDGF-A, PDGF-B, and VEGF gene expression in rats. AB - BACKGROUND: There is evidence from in vitro studies to suggest that the genes of platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) are, like the erythropoietin gene, regulated by oxygen tension. Hypoxia induced stimulation of, for example, PDGF or VEGF might be involved in the pathogenesis of acute or chronic renal failure and in renal 'inflammatory' diseases (glomerulonephritis, vasculitis, allograft rejection). METHODS: Male Wistar rats were exposed to chronic normobaric hypoxia (10% O(2), 90% N(2)) for 4 weeks. Additional groups of rats were treated with the endothelin receptor antagonist LU13525 and the NO donor molsidomine. Renal mRNA levels of PDGF-A, PDGF-B, and VEGF were semiquantitated using RNase protection assays. RESULTS: Renal gene expression of PDGF-A and PDGF-B was neither affected by 2 or 4 weeks of hypoxia nor by concomitant treatment with LU135252 or molsidomine. Chronic hypoxia did also not change VEGF gene expression; however, concomitant treatment with LU135252 increased all VEGF subtypes (188, 164, 120). CONCLUSIONS: The findings of the present study suggest that renal PDGF and VEGF gene expression in vivo during chronic hypoxia for 2 and 4 weeks is not sensitive to tissue hypoxia in contrast to cell culture experiments. During chronic hypoxia with concomitant blockade of endothelin receptors, all VEGF subtypes were increased, suggesting an inhibitory action of endothelins with regard to renal VEGF gene expression. PMID- 11014987 TI - Renal antioxidant enzymes and fibrosis-related markers in the rat adriamycin model. AB - Excessive generation of reactive oxygen intermediates can induce changes in the cellular antioxidant defence system. In this study we examine the antioxidant enzyme status and the expression of fibrosis-related marker proteins in the Adriamycin model of chronic renal failure in the rat. Twenty weeks after Adriamycin treatment, rats have overt nephrotic syndrome and renal failure with development of tubulo-interstitial fibrosis and glomerulosclerosis. Lipids accumulate in blood and in both glomeruli and tubulo-interstitial tissue. Desmin and alpha-smooth muscle actin expression increases in glomeruli and in the tubulo interstitial area. Renal cortex antioxidant enzyme activities are decreased 20 weeks after Adriamycin injection (to 41% for catalase, to 56% for total superoxide dismutase and to 69% for glutathione peroxidase). The mRNA levels of catalase, Cu/Zn-superoxide dismutase and glutathione peroxidase-1 evaluated by Northern blot are decreased by more than 50% for catalase, Cu/Zn-superoxide dismutase and glutathione peroxidase-1. We conclude that in the rat Adriamycin induced model of chronic renal failure with fibrosis, the combination of decreased antioxidant enzyme status in renal cortex with high concentrations of lipids in blood and renal tissue facilitates oxidative damage. Development of fibrosis is paralleled by increased expression of desmin and alpha-smooth muscle actin. PMID- 11014988 TI - Investigation of the localization of dehydroepiandrosterone sulphotransferase in adult rat kidney. AB - Sulphotransferases are a family of enzymes involved in the metabolism and detoxification of many compounds. Dehydroepiandrosterone (DHEA) sulphotransferase (DHEA-ST), which catalyzes the sulphation of steroids such as DHEA, is present in rat liver and adrenals. Sulphated steroids are present in urine, and many other enzymes which catalyze detoxification reactions are found in the kidney. There are not previous reports of DHEA-ST localization in adult kidney. The activity of DHEA-ST was investigated in adult rat kidney by a radio-isotope assay with DHEA as the substrate. Western blotting was used to assess protein expression, and the localization of DHEA-ST was investigated by immunohistochemistry. The DHEA-ST activity in rat kidney was found to be approximately four times less than that in rat liver. In female kidney, the activity was 1.46 +/- 0.06 nmol/min/microg, and in male kidney the activity was 1.29 +/- 0.09 nmol/min/microg. Investigation of protein expression gave a single band at 35 kDa which signified the presence of this enzyme in both male and female adult rat kidneys. Localization studies showed positive staining at high intensity in the collecting ducts of the medulla and in the S3 portion of the proximal convoluted tubule in the cortex. The distribution within the proximal tubules was restricted to the brush border. Reverse-transcriptase polymerase chain reaction showed DHEA-ST RNA expression in adult rat kidney and liver. The presence of this enzyme and its location in the kidney may suggest that in situ sulphation via DHEA-ST may play an important role in the excretion of endogenous and exogenous compounds. PMID- 11014989 TI - Proteinuria caused by lysozymuria mimics nephrotic syndrome. PMID- 11014990 TI - 'Isolated' thrombocytopenia by splenic sequestration in hemodialyzed patients. PMID- 11014992 TI - Pseudohypercreatininemia due to positive interference in enzymatic creatinine measurements caused by monoclonal IgM in patients with Waldenstrom's macroglobulinemia. PMID- 11014991 TI - Evidence for accumulation of advanced glycation end products in acute renal failure. PMID- 11014994 TI - Mutagen accumulated in cyst fluid of acquired cystic disease of the kidney. PMID- 11014993 TI - Streptococcus pneumoniae-induced acute renal failure in a neonate. PMID- 11014995 TI - Nephrotic syndrome and amyloid A amyloidosis in a patient with Erdheim-Chester disease. PMID- 11014996 TI - The influence of steroid therapy in patients with IgA nephropathy. PMID- 11014997 TI - Glomerular endothelial dysfunction and altered cytokines in severe nephrosis. PMID- 11014998 TI - Intravenous low-dose iron administration in hemodialysis patients treated with erythropoietin: 1-year follow-up study. PMID- 11014999 TI - Serum ferritin and oxidative stress in patients undergoing hemodialysis. PMID- 11015001 TI - What determines how satisfactory a size indicator is as a normalizing parameter for small solute clearances in peritoneal dialysis? PMID- 11015000 TI - Norfloxacin-induced acute interstitial nephritis. PMID- 11015002 TI - Antioxidative treatment retards progression of idiopathic membranous nephropathy. PMID- 11015003 TI - Effect of polymyxin B-immobilized fiber hemoperfusion on sepsis-induced rhabdomyolysis with acute renal failure. PMID- 11015004 TI - Hepatitis G virus markers in primary glomerulonephritis. PMID- 11015005 TI - An increased serum level of parathyroid hormone is a risk factor for aortosclerosis in hemodialysis patients. PMID- 11015006 TI - Influence of anemia on treatment of malnutrition in patients on hemodialysis. PMID- 11015008 TI - Serum KL-6 concentrations in hemodialysis patients with idiopathic interstitial pneumonitis. PMID- 11015007 TI - The concept of danger weight in intermittent peritoneal dialysis. PMID- 11015009 TI - IL-1 receptor antagonist production by isolated mononuclear cells is a better indicator of bioincompatibility of hemodialysis membranes than that by stimulated whole blood. PMID- 11015010 TI - r-HuEPO resistance and dialysate chloramine contamination in patients on hemodialysis. PMID- 11015011 TI - Do erythrocyte sedimentation rate and C-reactive protein levels have diagnostic usefulness in patients with renal failure? PMID- 11015012 TI - Increased urinary NAG excretion in hypertensives can decline with antihypertensive treatment. PMID- 11015013 TI - Allograft de novo membranous nephropathy complicating Alport's syndrome. PMID- 11015014 TI - Pyelonephritis caused by mesalazine. PMID- 11015015 TI - Tubulointerstitial changes are less important in membranoproliferative glomerulonephritis than in IgA nephropathy. PMID- 11015016 TI - Endothelial nitric oxide synthase gene polymorphism in patients with IgA nephropathy. PMID- 11015017 TI - Utility of serum and urinary transforming growth factor-beta levels as markers of diabetic nephropathy. PMID- 11015019 TI - Bilateral spontaneous perinephric hematoma: evaluation with CT and angiography and successful conservative treatment. PMID- 11015018 TI - Aldosterone influences serum magnesium in Gitelman syndrome. PMID- 11015020 TI - Renal cholesterol microembolism: is steroid therapy effective? PMID- 11015021 TI - Endothelial cell cytotoxicity and renal hypoperfusion in idiopathic nephrotic syndrome. PMID- 11015022 TI - Communication between acquired renal cysts and renal tubules. PMID- 11015023 TI - Expression of receptor for advanced glycation end product mRNA by human peritoneal mesothelial cells. PMID- 11015024 TI - Comparison between the angiotensin II receptor antagonist candesartan cilexetil and the angiotensin-converting enzyme inhibitor trandolapril in microalbuminuria of patients with early diabetic nephropathy. PMID- 11015025 TI - Low-density lipoprotein apheresis can improve type AA systemic amyloidosis. PMID- 11015026 TI - Prune-belly syndrome in a geriatric patient. PMID- 11015027 TI - Crescent formation in perimembranous-type renal amyloidosis associated with multiple myeloma. PMID- 11015028 TI - Depression and sleep disorders: clinical relevance, economic burden and pharmacological treatment. AB - A wide range of studies have been published over the past two decades that involve the intersection of sleep EEG, insomnia, psychiatric illness (especially depressive disorders) and psychopharmacology. Much of value has been discovered, but there have also been false starts and contradictory results. There is in fact strong evidence that insomnia is associated with medical and psychiatric illness and that the sleepiness associated with insomnia is the cause of many accidents. Thus, the direct (visits to doctors, cost of sleeping medication, complications from use of these medications) and indirect (accidents, quality of life) costs of insomnia are enormous and constitute a major public health problem in the industrialized countries. Believing that it is now timely to assess the state of this important research area, a consensus conference was convened on June 26-28, 1998, in Porto Cervo (Italy) to attempt to clarify the important issues and findings on the clinical effect of the different classes of antidepressant drugs on sleep quality in depression. The participants' consensus on some of the main topics is presented with the hope that this discussion and analysis will contribute to productive research in this important field. PMID- 11015029 TI - Effects of antidepressants on gamma-aminobutyric acid- and N-methyl-D-aspartate induced intracellular Ca(2+) concentration increases in primary cultured rat cortical neurons. AB - We investigated the effects of antidepressants on the intracellular Ca2+ concentration ([Ca2+]i) increases induced by gamma-aminobutyric acid (GABA) or N methyl-D-aspartate (NMDA) in primary cultured rat cortical neurons using fluorescence imaging. Acute treatment with imipramine inhibited GABA- and NMDA induced increases in [Ca2+]i in a concentration-dependent manner. Doses of 30 microM clomipramine, desipramine, amoxapine and maprotiline also inhibited both the GABA- and NMDA-induced [Ca2+]i increases significantly. Both inhibitory effects of the five major antidepressants on the GABA- or the NMDA-induced [Ca2+]i increases were well-correlated. Imipramine could inhibit significantly high-K+-induced [Ca2+]i increases. Our previous study has already shown that the GABA-induced [Ca2+]i increase involves a similar pathway to high-K+-induced Ca2+ influx. In conclusion, imipramine and several other antidepressants have acute inhibitory effects on the GABA-, NMDA- and high-K+-induced [Ca2+]i increases, suggesting that these inhibitory effects are not related to specific receptors. One possibility is that these effects may be commonly mediated via part of the high-K+-induced [Ca2+]i pathway. PMID- 11015031 TI - Effect of paroxetine on thyroid hormone levels in severely depressed patients. AB - There are very divergent appraisals of the effect of antidepressants on thyroid parameters and their possible correlation with the response. Whereas there are numerous investigations of tricyclic antidepressants, so far, there are only limited data on the possible effect of serotonin-selective reuptake inhibitors on neuroendocrine parameters. The present study showed a significant reduction of 11. 2% in thyroxine during treatment with 20 mg paroxetine in 25 severely depressed patients. PMID- 11015030 TI - Tissue-specific regulation of the peripheral benzodiazepine receptor by antidepressants and lithium. AB - Peripheral benzodiazepine receptors (PBR) have been identified in peripheral organs as well as in brain glial cells. PBR differ from central benzodiazepine receptors (CBR) in their lack of coupling to the gamma-aminobutyric acid receptors and the chloride ion channels. We investigated the effect of 21 days administration, followed by 7 days withdrawal, of fluvoxamine (10 mg/kg), desipramine (10 mg/kg) and lithium carbonate (25 mg/kg) on PBR and CBR binding characteristics in male Sprague-Dawley rats. All three agents significantly increased PBR density in the testes and adrenals. All tested drugs induced a significant decrease in PBR density in the kidney and liver. After withdrawal, PBR density remained decreased in the liver in all three groups and in the kidneys of the desipramine- and lithium-treated animals. In the cerebral cortex, CBR density increased in response to all three agents, whereas PBR density decreased significantly in response to desipramine and lithium carbonate. Chronic treatment with fluvoxamine, desipramine and lithium carbonate is apparently associated with a modulation in PBR expression in the testes, adrenals, kidneys, liver and brain, and in CBR expression in brain. The relevance of these tissue selective alterations to the antidepressive and/or anxiolytic effects of these agents, or their adverse effects, still needs to be determined. PMID- 11015032 TI - Zolpidem and promethazine in pre-anaesthetic medication. A pharmacopsychological approach. AB - This study dealt with the question: What are the effects of different doses of the benzodiazepine-like agent zolpidem and the phenothiazine derivative promethazine on mood in pre-anaesthetic medication? Subjects were 192 female and male patients awaiting elective surgery. Two drugs were administered to the patients in the evening before anaesthesia according to a 3 x 2 factorial randomized double-blind design. The first drug factor was zolpidem (8.03 vs. 16.06 mg) versus placebo. These drugs were combined with the second drug factor, either promethazine (50 mg) or placebo. In summary, the results confirm the hypothesis that zolpidem 8.03 mg can induce paradoxical effects, especially of anxiety, 1 h after application. When the dosage is doubled, no adverse effects are found, but there are no effects different from placebo either. Men could probably be described as a risk population for the adverse effects of zolpidem 8.03 mg. Furthermore, selective deactivating effects of promethazine 50 mg were shown in this study. In addition to these anaesthesiological aspects, this study showed promethazine to be useful as a research tool, suitable as a reference drug to detect deactivating effects. The multidimensional self-report inventory BSKE(EWL) (Befindlichkeitsskalierung durch Kategorien und Eigenschaftsworter) detected the expected drug effects, indicating that this instrument is suitable for further pharmacopsychological research. PMID- 11015033 TI - Electrophysiologic analysis of antidepressant drug effects on the GABA(A) receptor complex based upon antagonist-induced encephalographic power spectrum changes. AB - To better understand antidepressant drug effects on the GABA(A) receptor complex (the GABA(A) receptor, chloride ionophore and benzodiazepine receptor), we investigated how antidepressants influenced power spectrum changes induced by pentylenetetrazol (PTZ), a chloride ionophore antagonist, in the rat hippocampal electroencephalogram (EEG). In control recording, PTZ (27.5 mg/kg i. p.) increased EEG power at frequencies under 12 Hz up to five times. After rats were pretreated with imipramine, fluoxetine or trazodone for 7 days (10 mg/kg i.p., twice a day), PTZ could not increase EEG power to more than three times the power before injection; this effect was not observed after pretreatment for 3 days. These three antidepressants inhibit serotonin uptake, while two other antidepressants, desipramine and nortriptyline, that inhibit norepinephrine uptake failed to counter the PTZ effect. We concluded that antidepressants with serotonergic effects enhanced the function of the GABA(A) receptor complex. PMID- 11015034 TI - Effects of diazepam on auditory evoked potentials of rats elicited in a ten-tone paradigm. AB - The effect of diazepam on sensory gating was studied in rats by measuring diazepam effects on auditory evoked potentials (AEPs) elicited in a ten-tone paradigm. Trains of 10 repetitive tone-pip stimuli were presented. Rats (n = 8) received 4 mg x kg(-1) diazepam subcutaneously or vehicle, counterbalanced over two sessions. Diazepam decreased the amplitude of the middle-latency P30 component and increased the amplitudes of the late-latency N60 and P67 components. The increase in the late-latency components might be due to a diazepam-induced decrease in arousal. Stimulus repetition decreased the amplitudes of the middle-latency N18 and P30 components in both conditions. This suggests that automated neuronal recovery functions underlying sensory gating remain intact with diazepam. In the vehicle condition, the amplitude of the late latency P67 decreased with stimulus repetition, but not in the diazepam condition. This suggests a diazepam-induced decrease of behaviourally mediated habituation. PMID- 11015035 TI - Regulatory mechanisms in the retinal and choroidal circulation. AB - The retina receives its nutrients from two separate circulations: retinal and choroidal circulation. This short overview describes the determinants in the regulation of these circulations. Retinal circulation is characterized by a low blood flow while flow in the choroid is high. The choroidal circulation is mainly controlled by sympathetic innervation and is not autoregulated. Retinal circulation lacks autonomic innervation, shows an efficient autoregulation and is mainly influenced by local factors. Local mediators released by endothelial cells and surrounding retinal tissue also have a substantial role in the regulation of retinal circulation. PMID- 11015036 TI - nm23 Expression in choroidal melanoma. AB - nm23 protein expression of choroidal melanoma was investigated to determine its relationship with clinical and histopathological characteristics of the tumour. Thirty-four consecutive choroidal melanoma patients were examined by immunohistochemistry. Although age, sex, tumour cell type, tumour size, pigmentation, necrosis, apoptosis and tumour lymphocytic infiltration were not correlated with nm23 protein expression, tumours with low percentages of nm23 positive cells revealed higher nuclear grades and predominant mitotic figures. nm23 may be associated with melanoma progression, but there is no proof that it plays a role in the metabolic process of the tumour. PMID- 11015037 TI - The suppressive effect of tecogalan sodium on in vitro angiogenesis via the periendothelial proteolytic activities. AB - The exact mechanisms of the angiostatic effect by tecogalan sodium (TS) remain unclear. We examined the effects of TS on in vitro angiogenic activity, proteolytic activity and proliferation of retinal vascular endothelial cells (RECs). TS markedly inhibited the in vitro angiogenic activity of RECs although the growth inhibition of RECs was small. TS apparently decreased the cell associated urokinase-type plasminogen activator (uPA) activity and matrix metalloprotease 1 (MMP-1) activity even in the presence of anti-bFGF IgG. Thus, the suppression of the periendothelial matrix-degrading activities related to uPA and MMP-1 is suggested to be another possible mechanism of the antiangiogenic effect of TS, besides its prevention of bFGF REC binding which has previously been reported. PMID- 11015038 TI - Immobilization stress induces elevation of intraocular pressure in rabbits. AB - The aim of this study was to test whether immobilization and intravenous volume load as stressors influence the intraocular pressure in rabbits. Rabbits were immobilized for 1 h in a horizontally placed plastic tube with an internal diameter of 13.2 cm and a length of 33.2 cm. After immobilization, rabbits received rapid intravenous drip infusion of 5% glucose solution, 20 ml/kg of body weight, in 5 min. The intraocular pressure immediately after immobilization (11.2 +/- 3.0 mm Hg; mean and standard deviation) was significantly higher compared to control rabbits without immobilization (9.2 +/- 1.0 mm Hg, Student's t test, p = 0.0462). This difference became significantly larger when volume load was exerted on both groups of rabbits (22.7 +/- 5.6 versus 16.4 +/- 2.2 mm Hg, p = 0.0067). Serum levels of cortisol, adrenaline, and noradrenaline were significantly elevated after immobilization (p = 0.0002, p = 0.0271, p = 0.0296, respectively). Venous pressure of the ear tended to increase in rabbits immediately after immobilization (15.8 +/- 3.1 mm Hg) compared with control rabbits (8.5 +/- 2.3 mm Hg), and the difference became significant when volume load was exerted on both groups of rabbits (20.8 +/- 7.4 versus 9.2 +/- 4.8 mm Hg, p = 0.0211). In conclusion, we clearly demonstrated that physical stress due to immobilization, especially in combination with volume load, increased intraocular pressure in rabbits. PMID- 11015039 TI - Development of a completely encapsulated intraocular pressure sensor. AB - A completely encapsulated intraocular pressure (IOP) sensor equipped with telemetric signal and energy transfer is introduced integrated into a silicone disc for implantation into the eye. After implantation into enucleated pig eyes and into rabbit eyes in vivo, the IOP was recorded and compared to established techniques of IOP measurement. Pressure chamber tests showed that the sensor functioned correctly after biocompatible encapsulation in polydimethylsiloxane. In vivo and in vitro tests in rabbit and pig eyes demonstrated that the implanted system worked with the same precision as established techniques for IOP determination. The correlation between the measurements with the implanted device and pneumotonometry in several experiments was between 0.9 and 0.99. This device serves as a functioning model for the realization of a telemetric IOP sensor for integration into an artificial intraocular lens. Such a device will open new perspectives, not only in the management of glaucoma, but also in basic research for mechanisms of glaucoma. PMID- 11015040 TI - Cholesterol content of focal opacities and multilamellar bodies in the human lens: filipin cytochemistry and freeze fracture. AB - Adult human lenses with focal opacities were processed for normal as well as freeze fracture electron microscopy. Cholesterol was demonstrated using filipin cytochemistry. Filipin cytochemistry in combination with freeze fracture revealed that the amount of cholesterol in the normal fibre membranes was fully comparable with the amount of cholesterol in the deviating membranes of the focal opacities and the multilamellar bodies. As regards the cholesterol/phospholipid ratio, the deviating membranes have the same state of maturation as the surrounding normal lens fibre membranes, which implies that their synthesis is as highly coordinated as the synthesis of the normal lens fibre membranes. PMID- 11015041 TI - Universal opacity standard for Scheimpflug photography. AB - PURPOSE: To develop a universal standard for comparison of the results from different studies using Scheimpflug photography. MATERIAL AND METHOD: A suspension including polystyrene spheres was selected as the universal standard. The polystyrene suspension was diluted into 10 concentrations by adding spectroscopic quality water. An artificial eye chamber was designed to hold the diluted suspension for photography. Each concentration was photographed by both a Topcon SL-45 slit-lamp camera and a Nidek EAS-1000 system. The intensity of back scattered light was analyzed statistically. RESULTS: The relationship between the concentration of the polystyrene suspension and the internal gray scale step within the Topcon SL-45 camera can be described by a second-order polynomial. The relationship between the concentration of the polystyrene suspension and the measurement of the Nidek EAS-1000 system can also be expressed by a second-order polynomial. The relationship between the Topcon SL-45 camera and the Nidek EAS 1000 system is linear. CONCLUSIONS: The results of lens nuclear opacity measured with different Scheimpflug cameras can be compared using the method established in the study. The linear relationship between the Topcon SL-45 and Nidek EAS-1000 systems can be used to convert measurements from one system to the other. PMID- 11015043 TI - Inside Washington. PMID- 11015042 TI - Serum methanol levels in subjects with or without optic nerve head disease. AB - We evaluated serum methanol levels in subjects with or without optic nerve head disease. Serum methanol levels were determined using gas chromatography in 71 patients with optic nerve head disease and in 127 subjects without optic nerve head disease. Their ages ranged from 17 to 89 years. Serum methanol levels in 127 subjects without optic nerve head disease ranged from 0.12 to 3.86 microg/ml (mean +/- standard deviation, 1.72 +/- 0.86 microg/ml). In the subjects without optic nerve head disease, the differences in the levels between those with cataract versus retinal detachment, men versus women, and between each age-group (50-80 years) were not significant. The methanol levels in patients with optic neuritis (n = 2), Wolfram syndrome (n = 1), Leber hereditary optic neuropathy at the late stage (n = 2), retinitis pigmentosa (n = 23), and primary open-angle glaucoma (n = 16) were less than 3.86 microg/ml. Methanol levels in 1 patient with Leber hereditary optic neuropathy at the acute stage was 5.28 microg/ml. Of 10 patients with primary angle-closure glaucoma, 1 had a slightly elevated level and 9 had levels less than 3.86 microg/ml. Of 17 patients with normal tension glaucoma, 5 had methanol levels higher than 3.86 microg/ml, and 12 patients had levels less than 3.86 microg/ml. The present study shows that serum methanol levels in subjects without optic nerve head disease ranged from 0.12 to 3.86 microg/ml and were much lower than the levels that produce acute ocular symptoms of methanol intoxication. It is possible that high serum methanol levels may play a part in the acute stage of Leber hereditary optic neuropathy and normal tension glaucoma in certain patients. It is unlikely that increased serum methanol levels participate in primary angle-closure glaucoma. PMID- 11015044 TI - People to People. PMID- 11015045 TI - Coming Meetings. PMID- 11015046 TI - Emergency nursing in perspective: mentoring, sharing, supporting. PMID- 11015047 TI - Growing old is hard to do! PMID- 11015048 TI - More on customer surveys run amok. PMID- 11015049 TI - More on customer surveys run amok. PMID- 11015050 TI - Experienced ED nurses have many options. PMID- 11015051 TI - Experienced ED nurses have many options. PMID- 11015053 TI - Experienced ED nurses have many options. PMID- 11015054 TI - Experienced ED nurses have many options. PMID- 11015055 TI - Zero waiting room time does not work when patients outnumber beds. PMID- 11015059 TI - Dietary supplement or dangerous drug? Two case reports of gamma-butyrolactone toxicity. PMID- 11015060 TI - Geriatric assessment in the emergency department. PMID- 11015061 TI - Elder neglect assessment in the emergency department. AB - INTRODUCTION: Emergency departments are often the first point of contact for elder neglect victims. The purpose of this article is to describe a pilot study pertaining to the screening of patients and detection of elder neglect conducted in a large metropolitan medical center emergency department. The research question to be answered was, "Is it feasible for ED nurses to conduct accurate screening protocols for elder neglect in the context of their busy practice?" METHODS: During a 3-week period, 180 patients older than age 70 years (90% of all possible elderly patients during the screening hours) were screened to determine if they met the study criteria and could be enrolled into the protocol. RESULTS: Thirty-six patients met the eligibility criteria to enroll in the study, and 7 patients screened positive for neglect by a home caregiver. The nurses were able to screen and detect elder neglect with more than 70% accuracy, confirming the research question. The true-positive rate was 71%, and the false-positive rate was 7%. DISCUSSION: Elder neglect protocols are feasible in busy emergency departments, and neglect can be accurately detected in the emergency department when screening procedures are in place. PMID- 11015062 TI - The millennial challenge: elder abuse. PMID- 11015063 TI - Falls in the elderly: a multifactorial problem. PMID- 11015064 TI - Teaching geriatric concepts to health care providers. PMID- 11015065 TI - Activities and procedures performed by nurse practitioners in emergency care settings. AB - INTRODUCTION: Although nurse practitioners (NPs) have been practicing in emergency care (EC) settings for at least 25 years, little is known about the activities and procedures they perform. METHODS: A questionnaire was sent by either by E-mail or US mail to a convenience sample of 96 subjects. These 96 NPs were instructed to duplicate the questionnaire and distribute it to other NPs they may know who also work in EC settings. The questionnaire contained 71 activities and procedures obtained from Clinical Procedures in Emergency Medicine by Roberts and Hedges. The NPs were asked to rate the 71 activities and procedures according to the frequency with which they performed them, where they learned to perform them, and how important they believe it is that NPs in EC settings know how to perform them. RESULTS: Seventy-two NPs in EC settings responded. Fifty percent (n = 36) or more had performed 35 of the 71 activities and procedures. Almost every NP (n = 71) had used fluorescein staining, and only 3 procedures--culdocentesis, venous cutdown, and insertion of pins for skeletal traction--had never been performed. The majority of NPs learned to perform each of the activities and procedures through on-the-job training and continuing education courses. Fifty percent or more identified 56 activities and procedures as being important for NPs to know how to perform in EC settings. DISCUSSION: The results of this study indicate that whereas 50% or more of the NPs in EC settings had performed 35 out of 71 activities and procedures, 50% or more indicated that a larger skill set of 56 activities and procedures is believed to be needed for practice. PMID- 11015066 TI - Seen but not heard: battered women's perceptions of the ED experience. AB - INTRODUCTION: A plethora of studies describe helping professionals' responses to and actions directed toward battered women in the emergency department. However, research that yields data regarding the clients' perceptions about their actual experiences in the ED setting is sorely needed. The aim of this study was to describe battered women's perceptions of their ED experience. METHOD: A qualitative design, namely a phenomenologic approach, was used for this inquiry to enable the women to express themselves in their own voices. Informants were recruited from shelters for battered women. Women who had sought help for abuse related injuries at a hospital emergency department within the past 12 months were asked to participate. Methods used to collect data were in-depth, individual, audiotaped interviews and demographic data sheets. Data analysis was conducted using Colaizzi's (1978) procedural steps. RESULTS: Several categories emerged as being descriptive of the women's perceptions of their ED experience. Themes identified included the women's feelings during the visit, such as fear of their partner, concern for children, and loneliness; the women's belief that the ED staff do not understand abuse; satisfaction with treatment of physical injuries but dissatisfaction with how the issue of abuse is managed; the difficulty of disclosing the abuse because of fear, embarrassment, and a lack of resources; and a request that health care professionals display compassion, provide referrals, and offer options. DISCUSSION: The women's narratives explicate their feelings during the ED visit and sensitize nurses to their experience. The reports of dissatisfaction with the care they received in the emergency department add to the validity of findings from previous studies that have documented similar results and point to the need to examine and reshape the delivery of care to abused women in the emergency department. PMID- 11015067 TI - Designing and implementing a computerized tracking system: the experience at one level I trauma center emergency department. PMID- 11015068 TI - Hospice care in the emergency department: important things to remember. PMID- 11015069 TI - Pediatric education for prehospital professionals. PMID- 11015071 TI - Take care. PMID- 11015072 TI - The Bowen case: who was the EMTALA villain--the doctor or the nurse, or neither? PMID- 11015073 TI - Developing an ED training program: how to "grow your own" ED nurses. PMID- 11015074 TI - Our new rapid treatment unit: an innovative adaptation of the "less than 24-hour stay" holding unit. PMID- 11015075 TI - Ergonomics, policy, and the ED nurse. PMID- 11015076 TI - The elderly rape victim: stereotypes, perpetrators, and implications for practice. PMID- 11015077 TI - Trauma case management defined. PMID- 11015078 TI - A 71-year-old man with syncope, respiratory distress, and hypotension. PMID- 11015081 TI - Presidential address: dead wrong--thinking about...thinking... about...health care. PMID- 11015082 TI - Academic practice groups: strategy for survival. AB - BACKGROUND: The mission of public academic health centers (puAHC) and their affiliated practice groups (APG) focuses on teaching, research, and the clinical care of at-risk populations. Resources to accomplish this mission, however, are becoming scarce. For puAHC to survive and remain competitive, innovative strategies will need to be developed by the APG. We hypothesized that the integration of a surgical academic practice of the APG with a nonacademic integrated health care delivery system (NAIDS) in a managed care environment would benefit all involved. METHODS: A surgical academic practice was integrated with a NAIDS in a 95% managed care market. Faculty alone provided care the first year, and third-year residents were added the following year. To assess outcome, we collected benefit and cost data for the 1-year period before integration and compared them with the two, 1-year periods after integration. RESULTS: In the second year of integration, revenues from the NAIDS referrals to the puAHC and APG increased 89% and 150%, respectively. The NAIDS' general surgical and endoscopy caseload increased by 25%. Additionally, there was a 92% reduction in operating room technician cost with no increase in operating time per case. Finally, the third-year resident experienced a caseload increase of 163%. CONCLUSIONS: In an environment where resources are diminishing and managed care consists of many large NAIDS that drive referrals and revenue, the integration of a surgical academic practice with a NAIDS benefits all shareholders. Academic practice groups that develop strategies that leverage their competitive advantage will have the best chance of surviving in today's turbulent health care market. PMID- 11015083 TI - Sustained infection induces 2 distinct microvascular mechanisms in the splanchnic circulation. AB - BACKGROUND: Altered intestinal blood flow during systemic inflammation leads to organ dysfunction. Mucosal ischemia occurs during sepsis despite an increase in portal blood flow. We hypothesized that separate mechanisms are active in the large resistance and small mucosal microvessels to account for this dichotomy. METHODS: Chronic infection was induced in rats by bacterial inoculation (Escherichia coli and Bacteroides fragilis) of an implanted subcutaneous sponge. Separate groups were studied at 24 and 72 hours after a single inoculation of bacterium or 24 hours after a second inoculation (ie, 72 hours of sepsis). Time matched controls were used for each group. Intravital microscopy of the terminal ileum was used to assess endothelial-dependent vasodilation to acetylcholine (10( 9) to 10(-5) mol/L) in resistance (A(1)) and premucosal (A(3)) arterioles. Threshold sensitivity (-log of 20% response dose) was calculated from dose response curves for each animal. RESULTS: Vasodilator sensitivity to acetylcholine in A(1) arterioles was significantly decreased at 24 hours, and these changes persisted up to 72 hours after a single bacterial inoculation. There was no change in the dilator sensitivity of A(3) arterioles after a single inoculation. When there was a challenge with a second bacterial inoculation, there was a reversal of the A(1) dilator response and an increase in A(3) sensitivity. CONCLUSIONS: An initial septic event results in a decrease in dilator reactivity in the resistance A1 arterioles that persists for at least 72 hours. A sustained septic challenge results in increased dilator reactivity in both A(1) and A(3) vessels. This enhanced sensitivity during sepsis suggests that more than 1 therapeutic approach to preservation of intestinal blood flow will be necessary. PMID- 11015084 TI - p21WAF1 expression is associated with improved survival after adjuvant chemoradiation for pancreatic cancer. AB - BACKGROUND: Cell cycle arrest after DNA damage is partly mediated through the transcriptional activation of p21(WAF1) by the p53 tumor suppressor gene. p21(WAF1) and p53 are both critical in maintaining cell cycle control in response to DNA damage from radiation or chemotherapy. Therefore, we examined the role of p21(WAF1) and p53 in the determination of outcome for patients who receive radiation and/or chemotherapy for pancreatic cancer. METHODS: p21(WAF1) and p53 protein expression were determined (with the use of immunohistochemistry) in specimens from 90 patients with pancreatic cancer. Forty-four patients underwent surgical resection, and 46 patients had either locally unresectable tumors (n = 9 patients) or distant metastases (n = 37 patients). Seventy-three percent of the patients who underwent resection and 63% of the patients who did not undergo resection received radiation and/or chemotherapy. RESULTS: p21(WAF1) expression was present in 48 of 86 tumors (56%) and was significantly (P<.05) associated with advanced tumor stage. Median survival among patients with resected pancreatic cancer who received adjuvant chemoradiation with p21(WAF1)-positive tumors was significantly longer than in patients with no p21(WAF1) staining (25 vs. 11 months; P = .01). Fifty of 89 tumors (56%) stained positive for p53 protein. p53 overexpression was associated with decreased survival in patients who did not undergo resection. CONCLUSIONS: Normal p21(WAF1) expression may be necessary for a beneficial response to current adjuvant chemoradiation protocols for pancreatic cancer. Alternate strategies for adjuvant therapy should be explored for patients with pancreatic cancer who lack functional p21(WAF1). PMID- 11015085 TI - A prospective surgical outcome study assessing the impact of parathyroidectomy on symptoms in patients with secondary and tertiary hyperparathyroidism. AB - BACKGROUND: To assess the impact of surgery on preoperative symptoms in secondary (2 degrees ) and tertiary (3 degrees ) hyperparathyroidism (HPT) compared with primary (1 degrees ) HPT. METHODS: Twenty-two patients with 2 degrees HPT and 10 with 3 degrees HPT were enrolled. Age-matched patients, 32 with 1 degrees HPT and 32 with thyroid disease were enrolled for comparison. An outcome questionnaire documented symptoms expressed as the median symptom index score (MSIS) preoperatively and at days 7 and 3 and 12 months postoperatively. RESULTS: Preoperatively, the MSIS for the groups with 3 degrees, 2 degrees, and 1 degrees HPT and thyroid disease was 225, 572, 372, and 146, indicating that patients with HPT were more symptomatic than those in the thyroid group (P<.05). Patients with 1 degrees HPT had a decrease in their MSIS at day 7 (195, P<.05) and at 3 and 12 months (159 and 156). Patients with 3 degrees HPT also had a decrease in their MSIS over time. Patients with 2 degrees HPT had a decrease in their MSIS at day 7 (469, P<.05); however, they remained more symptomatic at 3 and 12 months (410 and 355). CONCLUSIONS: Parathyroidectomy reduces many of the preoperative symptoms in HPT. Patients with 1 degrees and 3 degrees HPT have a similar resolution of their symptoms. Patients with 2 degrees HPT have an improvement in many of their symptoms, although they remain more symptomatic at 1 year. PMID- 11015086 TI - Surgical shunts and TIPS for variceal decompression in the 1990s. AB - BACKGROUND: In the 1990s, liver transplantations and transjugular intrahepatic portosystemic shunts (TIPS) have become the most common methods to decompress portal hypertension. This center has continued to use surgical shunts for variceal bleeding in good-risk patients who continue to bleed through endoscopic and pharmacologic treatment. This article reports this center's experience with surgical shunts and TIPS shunts from 1992 through 1999. METHODS: Sixty-three patients (Child A, 43 patients; Child B, 20 patients) received surgical shunts: distal splenorenal, 54 patients; splenocaval, 4 patients; coronary caval, 1 patient; and mesocaval, 4 patients. Sixty-two patients had refractory variceal bleeding, and 1 patient had ascites with Budd-Chiari syndrome. Two hundred patients (Child A, 24 patients; Child B, 62 patients; Child C, 114 patients) received TIPS shunts. One hundred forty-nine patients had refractory variceal bleeding, and 51 patients had ascites, hydrothorax, or hepatorenal syndrome. Data were collected by prospective databases, protocol follow-up, and phone contact. RESULTS: The 30-day mortality rate was 0% for surgical shunts and 26% for TIPS shunts; the overall survival rate was 86% (median follow-up, 36 months) for surgical shunts and 53% (median follow-up, 40 months) for TIPS shunts. For surgical shunts, the portal hypertensive rebleeding rate was 6.3%; the overall rebleeding rate was 14.3%. For TIPS shunts, the overall rebleeding rate was 25.5% (30-day, 9.4%; late, 22.4%). There were 4 reinterventions for surgical shunts (6.3%); the reintervention rate for TIPS shunts in the bleeding group was 33%, and the reintervention rate in the ascites group was 9.5%. Encephalopathy was severe in 3.1% of the shunt group and mild in 17.5%; this was not systematically evaluated in the TIPS shunts patients. CONCLUSIONS: Surgical shunts still have a role for patients whose condition was classified as Child A and B with refractory bleeding, who achieve excellent outcomes with low morbidity and mortality rates. TIPS shunts have been used in high-risk patients with significant early and late mortality rates and have been useful in the control of refractory bleeding and as a bridge to transplantation. The comparative role of TIPS shunts versus surgical shunt in patients whose condition was classified as Child A and B is under study in a randomized controlled trial. PMID- 11015088 TI - Evolution of sentinel lymph node biopsy for melanoma at a National Cancer Institute-designated cancer center. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) has rapidly evolved into the standard of care for clinically node-negative melanoma. Since adopting sentinel lymph node (SLN) technology in 1993, we have periodically reviewed our institution's results and made several modifications. METHODS: From January 1993 to December 1998, 182 patients with clinically node-negative primary cutaneous melanoma underwent SLNB. Charts were retrospectively reviewed and assessed for the technique for the identification of the SLN, the pathologic analysis, and the use of intraoperative frozen section. RESULTS: The accuracy of SLN identification improved from 91% to 100% with the combination of isosulfan blue dye and radiolabeled colloid over isosulfan blue dye alone. Routine versus selective lymphoscintigraphy identified 7 in-transit SLNs and increased detection of dual nodal basin drainage (15%-27%). Identification of micrometastases in the SLN increased from 14% to 24% after a modification of pathologic evaluation. The positive SLN was the only involved node in most patients (80%). Intraoperative frozen section had a sensitivity of 58% and was of benefit in only 13 of 124 patients (10%). CONCLUSIONS: Several modifications to the identification of the SLNs and the detection of metastatic melanoma have improved our outcome with SLNB. A careful, periodic review of results to identify areas for improvement at each institution is crucial to the success of SLNB for melanoma. PMID- 11015087 TI - Clinical outcomes and resource usage in 100 consecutive patients after off-pump coronary bypass procedures. AB - BACKGROUND: Cardiopulmonary bypass initiates a cascade of inflammatory processes that may result in end-organ damage, leading to the increased prevalence of noncardiac complications. Therefore, off-pump coronary artery bypass graft (OP CAB) procedures have recently been introduced into clinical practice. METHODS: This study was a case-controlled study that compared the outcomes and cost of 100 consecutive OP-CAB procedures with a control group of 100 contemporary matched conventional coronary artery bypass grafting procedures. All operations were performed by a single surgeon (J.H.L. ) and complete revascularization that used off-pump techniques was achieved with the use of innovative exposure techniques to the lateral and posterior wall vessels. RESULTS: An average of 3.1 grafts per patient were performed in the OP-CAB group (range, 1-5). The incidence of conversion to conventional coronary artery bypass grafting was 1%. The overall mortality rate was 2.0%. There were no instances of stroke, renal failure, or sternal infections in the OPCAB group. Thus, the OP-CAB group had a shorter length of stay (6.1+/-2.5 versus 7.1+/-3.3 d; P =.003), with a corresponding reduction in variable direct cost per case of 29% (P<.001). CONCLUSION: Our experience suggests that OP-CAB procedures are feasible for most patients who currently require complete revascularization. It is associated with very a low morbidity rate and may represent the ideal revascularization strategy for patients at high risk for undergoing cardiopulmonary bypass. PMID- 11015089 TI - Neoadjuvant chemotherapy, radical resection with intraoperative radiation therapy (IORT): improved treatment for gastric adenocarcinoma. AB - BACKGROUND: Adenocarcinoma of the stomach and gastroesophageal junction results in substantial morbidity, locoregional recurrence, and death. Surgical procedures, even with adjuvant therapy, have not significantly improved survival. This study evaluated the toxicity, response rate, locoregional control, and survival of patients with locally advanced gastric cancer that was treated with neoadjuvant multimodality therapy. METHODS: Patients with stage IIIA or early stage IV gastric adenocarcinoma received neoadjuvant 5-fluorouracil, Leucovorin, Adriamycin, and Cisplatin and underwent gastrectomy or esophagogastrectomy with intraoperative radiotherapy (IORT; 1000 cGY) to the gastric bed and postoperative radiation therapy. RESULTS: Nine of 15 patients (60%) with transmural extension and/or nodal metastases received IORT. There were 2 pathologically complete responses at the primary site. Eleven of 15 patients (73%) had tumor in perigastric lymph nodes; however, 9 of 15 patients (60%) had mucin-filled nodes without tumor cells. Neoadjuvant treatment did not increase operative morbidity rates. Ten of 15 patients (67%) remain free of disease (median, 27 months; range, 6-60 months). Five patients died 13 to 41 months (median, 17 months) after diagnosis. CONCLUSIONS: Neoadjuvant multimodality therapy with neoadjuvant 5 fluorouracil, Leucovorin, Adriamycin, and Cisplatin, radical resection with IORT, and postoperative radiation therapy is safe, can downstage tumors, provides improved locoregional control, and appears to cause significant tumor regression that may result in long-term survival or cure. PMID- 11015090 TI - Abdominal vena caval injuries: outcomes remain dismal. AB - BACKGROUND: The mortality rate for abdominal vena caval injuries remains high. We examined the experience of a level I trauma center to determine factors significant to the outcome in these injuries. METHODS: Forty-seven patients were identified in a retrospective review (1989 to 1999) of patients were identified with abdominal vena caval injury. Data were analyzed by uni- and multivariate methods, including logistic regression. RESULTS: Most of the individuals with abdominal vena caval injuries were young male patients who were injured by penetrating trauma and who were hypotensive on arrival. The severity of injury and the number of organs injured was high. The overall mortality rate was 55%. Nonsurvivors were more often hypotensive in the field with physiologic derangement consistent with hemorrhagic shock. Type and location of injury as well as method of repair were associated with death. Multiple regression analysis revealed that prehospital initial systolic blood pressure and intraoperative bicarbonate levels were independent predictors of survival. CONCLUSIONS: We identified factors related to poor outcome, including suprarenal and retrohepatic location of injury and variables that reflected the evolution of shock. Management should include appropriate resuscitation and ultimately may require novel operative techniques. PMID- 11015091 TI - Liver transplantation from controlled non-heart-beating donors. AB - BACKGROUND: The use of organs from non-heart-beating donors (NHBDs) has been proposed as one way to increase the donor pool. However, few centers have transplanted livers from NHBDs. We report here the results of 19 liver transplants from controlled NHBDs. METHODS: From January 1993 through August 1999, 364 liver transplantations were performed from heart-beating donors (HBDs) and 19 liver transplantations were performed from NHBDs. Donor and recipient characteristics, posttransplant complications, and patient and allograft survival were compared. RESULTS: No differences in hepatic artery, portal vein, or biliary complications were noted between the groups. However, the rate of primary nonfunction was higher in recipients of livers from NHBDs (10.5% vs. 1.3%; P = .04). No difference in patient survival was seen between recipients of NHBDs or HBDs (72.6% vs. 84.8%; P =.36); however, allograft survival was lower in recipients who received livers from NHBDs (53.8% vs. 80.9%; P =.007). CONCLUSIONS: Liver transplantation from controlled NHBDs results in similar patient survival and post-transplant complications. However, primary nonfunction was higher and allograft survival was less in recipients of livers from NHBDs. The results of liver transplantation from controlled NHBDs are encouraging and should continue to be cautiously pursued as one way to help alleviate the current shortage of donor livers. PMID- 11015092 TI - Efficacy of the laparoscopic approach for anterior lumbar spinal fusion. AB - BACKGROUND: This study compares the immediate postoperative outcomes in patients who undergo laparoscopic and open anterior lumbar spinal fusion and describes the learning curve associated with the performance of this procedure. METHODS: The charts of patients who underwent anterior lumbar spinal fusion between January 1995 and July 1999 were reviewed. Data pertaining to the operation and postoperative course were analyzed and compared. RESULTS: Eighty-nine patients underwent anterior lumbar spinal fusion. Fourteen patients were excluded; a full analysis was performed on the records of the remaining 75 patients. Fifty-five patients underwent an attempted laparoscopic procedure, and 20 patients underwent an open procedure. The conversion rate was 38% (21/55 patients) in the group who underwent the laparoscopic procedure. In the 34 patients whose laparoscopic procedure was completed, there was significantly less blood loss and shorter postoperative ileus, but the operative time was longer, when compared with patients who underwent the open procedure. The laparoscopic procedures performed in 1999 resulted in fewer conversions, less blood loss, and a shorter operating room time, when compared with the laparoscopic procedures in 1998. CONCLUSIONS: Laparoscopic anterior lumbar spinal fusion improves immediate postoperative results when compared with open anterior lumbar spinal fusion. PMID- 11015093 TI - Surgical treatment of anorectal complications in Crohn's disease. AB - BACKGROUND: The purpose of our study was to elucidate features, surgical procedures, and long-term results in patients with anorectal complications of Crohn's disease. METHODS: Physical findings, surgical treatment, and long-term outcome were recorded prospectively for 224 patients who had anorectal complications of Crohn's disease between October 1984 and May 1999. RESULTS: Presenting complications included abscess (n = 36), fistula-in-ano (n = 51), rectovaginal fistula (n = 20), anal stenosis (n = 40), anal incontinence (n = 11), or a combination of features (n = 66). Twenty-four patients did not undergo surgical treatment; the remaining 200 patients underwent 284 procedures. Ultimately, 139 patients (62%) retained anorectal function; reasons for proctectomy in the remaining 85 patients included disease (n = 66), extensive fistular disease (n = 15), fecal incontinence (n = 2), and tight anal stenosis (n = 1). Patients with rectal disease had a significantly higher rate of proctectomy than patients with rectal sparing (77.6% vs. 13.6%, respectively, P<.0001). In the absence of rectal involvement, patients with multiple complications had a significantly higher rate of proctectomy than patients with single complications (23% vs. 10%, P<.05). CONCLUSIONS: A wide spectrum of surgical techniques is required for the management of the diverse anorectal complications of Crohn's disease. Complete healing and control of sepsis can be achieved in the majority of patients. Active rectal disease and multiple complications significantly increase the need for proctectomy. PMID- 11015094 TI - Primary gastrointestinal sarcomas: analysis of prognostic factors and results of surgical management. AB - BACKGROUND: This study was done to review the clinical presentation, surgical management, and prognostic factors for primary gastrointestinal sarcomas. METHODS: We reviewed medical records of 55 patients who were treated for primary gastrointestinal sarcomas from 1981 through 1996. Mean follow-up time was 32 months. RESULTS: Clinical findings included gastrointestinal bleeding (51%), palpable mass (36%), and abdominal pain (33%). The stomach was the most common site of disease (53%), followed by the small intestine (33%). Tumors were high grade in 76% of patients and low-grade in 24% of patients. Complete resection of all gross disease was accomplished in 35 patients (64%), incomplete resection in 17 patients (31%), and biopsy only in 3 patients (5%). Adjacent organ resection was required in 19 patients (35%). Overall actuarial survival was 22% (median survival, 32 months). Unfavorable prognostic factors were incomplete resection, high-grade histologic features, and tumor size of 5 cm or more (P<.05). En bloc resection of contiguous organs did not adversely effect survival. In patients with complete resections, tumor grade was the most important prognostic factor (median survival, 55 months vs 19 months for low-grade vs high-grade tumors; P<.05). CONCLUSIONS: Aggressive surgical resection, including en bloc resection of locally advanced tumors, appears warranted. Despite complete resections, patients with high-grade tumors remain at risk for recurrence. PMID- 11015095 TI - Evaluating surgical competency with the American Board of Surgery In-Training Examination, skill testing, and intraoperative assessment. AB - BACKGROUND: Evaluation of surgical competency should include assessment of knowledge, technical skill, and judgment. The purpose of this study was to determine the relationship between the American Board of Surgery In-Training Examination (ABSITE), skill testing, and intraoperative assessment. METHODS: Postgraduate year 2 (PGY-2) and postgraduate year 3 (PGY-3) surgery residents (n = 33) were tested by means of (1) the ABSITE, (2) skill testing on a laparoscopic video-trainer, and (3) intra-operative global assessments during laparoscopic cholecystectomy. The Pearson correlation was used to determine the correlation between the ABSITE, skill testing, and intraoperative assessments. For the comparison of PGY-2 and PGY-3 resident performance, Wilcoxon rank sum tests were used. RESULTS: The ABSITE scores did not correlate with skill testing or intraoperative assessments (not significant). Skill testing correlated with the intraoperative composite score and with 4 of 8 operative performance criteria (P<.05). The ABSITE scores and skill testing were not different for PGY-2 and PGY 3 residents (not significant). Intraoperative assessments were better in 5 of 8 criteria and the composite score for PGY-3 versus PGY-2 residents (P<.05), which demonstrated construct validity. CONCLUSIONS: The ABSITE measures knowledge but does not correlate with technical skill or operative performance. Residency programs should use multiple assessment instruments to evaluate competency. There may be a role for both skill testing and intraoperative assessment in the evaluation of surgical competency. PMID- 11015096 TI - A 32-year experience in 100 patients with giant paraesophageal hernia: the case for abdominal approach and selective antireflux repair. AB - BACKGROUND: Giant paraesophageal hiatal hernia (GPEH) presents a risk of catastrophic complications that include massive bleeding, strangulation, and perforation and should be repaired. Controversy persists as to the surgical approach and whether an antireflux repair is required. METHODS: This study reviews the experience with 100 patients with GPEH who underwent surgical repair between 1967 and 1999. Eighty patients underwent an elective operation, and 20 patients underwent an emergency procedure for complications of GPEH. The gastroesophageal junction was above the hiatus ("combined" hernia with sliding component) in 23 patients and in the abdomen in 77 patients, including 3 patients with a true parahiatal hernia. RESULTS: A thoracic approach was used in 18 patients, mostly early in our experience; postoperative gastric volvulus requiring transabdominal repair developed in 2 patients. The remaining 82 patients underwent an abdominal repair, with temporary gastrostomy to prevent gastric displacement in 75 patients; the hernial sac was resected, and the hiatus was reconstructed in all of the patients. Thirty-five patients with reflux on preoperative work up underwent a fundoplication, with gastroplasty in 2 patients because of a short esophagus. No patient has experienced hernia recurrence. Whereas symptomatic relief was excellent in all patients with elective repair, mild reflux was present in 2 patients after emergency operation. There were no deaths among the patients who underwent elective operation; there were 2 hospital deaths among those patients who underwent emergency operation (10%). CONCLUSIONS: GPEH should be repaired soon after recognition. Reflux should be evaluated before the operation, and if present, fundoplication should be part of the repair along with the reduction of the hernia, excision of the sac, gastropexy, and crural closure. These are best achieved with an abdominal approach. PMID- 11015097 TI - The risk assessment profile score identifies trauma patients at risk for deep vein thrombosis. AB - BACKGROUND: The identification of trauma patients at risk for the development of deep venous thrombosis (DVT) at the time of admission remains difficult. The purpose of this study is to validate the risk assessment profile (RAP) score to stratify patients for DVT prophylaxis. METHODS: All patients admitted from November 1998 thru May 1999 were evaluated for enrollment. We prospectively assigned patients as low risk or high risk for DVT using the RAP score. High-risk patients received both pharmacologic and mechanical prophylaxis. Low-risk patients received none. Surveillance duplex Doppler scans were performed each week of hospitalization or if symptoms developed. Hospital charges for prophylaxis were used to determine the savings in the low-risk group. Statistical differences between the risk groups for each factor of the RAP and development of DVT were determined by the chi-squared test, with significance at a probability value of less than .05. RESULTS: There were 102 high-risk (64%) and 58 low-risk (36%) individuals studied. Eleven of the high-risk group (10.8%) experienced the development of DVT (asymptomatic, 64%). None of the low-risk group was diagnosed with DVT. Five of the 16 RAP factors were statistically significant for DVT. Eliminating prophylaxis and Doppler scans in low-risk patients resulted in a total savings of $18,908 in hospital charges. CONCLUSIONS: The RAP score correctly identified trauma patients at increased risk for the development of DVT. Despite prophylaxis, the high-risk group warrants surveillance scans. Withholding prophylaxis in low-risk patients can reduce hospital charges without risk. PMID- 11015098 TI - Management of biliary tract disease in heart and lung transplant patients. AB - BACKGROUND: Preexisting gallstones and pharmacologic alterations in both bile lithogenicity and immune function may predispose organ transplant recipients to the complications of biliary calculi. METHODS: Records of all 178 patients undergoing heart, lung, or heart-lung transplantation at our institution between 1980 and 1998 were reviewed. Patients with biliary tract disease were grouped as follows: group I, pretransplantation diagnosis and treatment; group II, pretransplantation diagnosis and posttransplantation treatment; group III, normal pretransplantation biliary tree with posttransplantation diagnosis and treatment; group IV, unknown pretransplantation biliary status with posttransplantation diagnosis and treatment. Comparison among groups was made with regard to ultrasound findings, presentation, indication for operation, procedure, and outcome. RESULTS: Of the 141 patients undergoing pretransplantation and/or posttransplantation ultrasound surveillance, the prevalence of abnormal ultrasonography was 36%. All patients in group I (n = 11) underwent elective intervention without complication. Of the 14 patients (groups II through IV) undergoing posttransplantation operation, intervention was mandated by acute complications of biliary tract disease in 7. The mortality rate in these 7 patients was 29%. CONCLUSIONS: Cholecystectomy in the posttransplantation period is often required emergently and has a high mortality. Posttransplantation surveillance of the biliary tree is crucial because of the high rate of de novo stone formation. All biliary calculi should be eradicated electively in stable patients before transplantation and on diagnosis after transplantation. PMID- 11015099 TI - All-artery multigraft coronary artery bypass grafting with only internal thoracic arteries possible and safe: a randomized trial. AB - BACKGROUND: The internal thoracic artery (ITA) bypass to the left anterior descending coronary artery is of proven benefit in multigraft coronary artery bypass. Total ITA grafts, if reoperation is averted by avoiding saphenous vein grafts (SVGs), are attractive. The safety of the total ITA graft operation (all ITA) is a concern. METHODS: A randomized trial of multiple-ITA bypass graftings with the use of bilateral sequential ITA without SVGs was performed. Control patients received 1 ITA plus SVG. Inclusion criteria were those used in the Coronary Artery Surgery Study, extended to age 76 years, and any angina class, except emergent. One hundred sixty-two patients were randomized (81 patients per group) from January 1, 1990, to December 31, 1994. RESULTS: Baseline traits were similar as were cross-clamp times, pump times, and number of arteries bypassed (average, 4.3 arteries). Patients who received multiple ITA grafts had no myocardial infarctions, per reference laboratory. One patient died, and 2 patients returned for bleeding. The ITA-SVG group had similar results. The all ITA group experienced successful completion in 93% of cases. Complications did not differ from control patients. CONCLUSIONS: Early and 5-year outcomes were not different between the all-ITA group and the ITA with SVGs group. We believe experienced surgeons can safely extend the ITA to multibypass coronary artery bypass without use of SVG to achieve an all-ITA operation. PMID- 11015100 TI - Laparoscopic splenectomy: outcomes and lessons learned from over 200 cases. AB - BACKGROUND: In this study of laparoscopic splenectomy (LS), we evaluate prospectively gathered perioperative patient data and review lessons learned in the evolution of this procedure. METHODS: At 2 university medical centers between November 1993 and March 2000, there were 203 patients (122 female patients and 81 male patients) who underwent LS after preoperative evaluation. RESULTS: LS was successfully completed in 197 patients (97%). The mean operative time was 145.5 minutes and the length of stay averaged 2.7 days with 143 (70.4%) staying less than 48 hours. The most common indication was idiopathic thrombocytopenic purpura (ITP). Six patients required conversion to open splenectomy (OS), with only 2 conversions in the last 163 cases. No deaths were attributed to the procedure. Complications occurred in 19 patients (9.3%). Thirty accessory spleens were identified in 25 patients (12.3%). Seventeen patients (8.4%) underwent concomitant procedures, most commonly cholecystectomy. CONCLUSIONS: LS by the lateral approach is both safe and feasible in patients of all ages. PMID- 11015101 TI - Long-term results of surgical repair of bile duct injuries following laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy (LC) is associated with an increased incidence of bile duct injuries when compared with the open surgical technique. Long-term results of repaired injuries and hepatic damage associated with chronic biliary obstruction are lacking. METHODS: From Aug 1, 1991 until Dec 1, 1999, there were 27 patients referred for management of complex biliary injuries that occurred during LC. Patients underwent percutaneous transhepatic cholangiography and placement of transhepatic catheters with computed tomography-guided biloma drainage when indicated. On the basis of the cholangiography findings, patients underwent Roux-en-Y hepaticojejunostomy (HJ) and liver biopsy or were treated with nonsurgical interventions. RESULTS: Twenty-one of 27 patients (77. 8%) underwent HJ, and 16 of these 21 patients (76.2%) also underwent hepatic biopsy. In 1 patient, a recurrent stricture developed at 20 months after the initial repair; and, in a second patient, an episode of cholangitis developed in the postoperative period with the transhepatic catheters in place. Five of 16 patients (31.2%) demonstrated marked hepatic fibrosis with 4 (25%) of these patients showing evidence of evolving cirrhosis at the time of HJ. CONCLUSIONS: In this series with 55 months of follow-up, HJ repair of LC injuries was associated with an initial 95.2% success rate and an ultimate success rate of 100%. Despite this, delayed referral, averaging 12 months, was associated with significant hepatic injury in 5 of 16 (31.3%) patients who underwent biopsy. PMID- 11015102 TI - Dynamic helical computed tomography scan accurately detects hemorrhage in patients with pelvic fracture. AB - BACKGROUND: The purpose of this study was to evaluate the use of dynamic helical computed tomography (CT) scan for screening patients with pelvic fractures and hemorrhage requiring angiographic embolization for control of bleeding. METHODS: Patients admitted to the trauma service with pelvic fractures were identified from the trauma registry. Data retrieval included demographics, hemodynamic instability, Injury Severity Score, blood transfusion requirement, length of stay, and mortality. CT scans obtained during the initial evaluation were reviewed for the presence of contrast extravasation and correlated with angiographic findings. Data are reported as mean +/- SEM, with P<.05 considered significant. RESULTS: Seven thousand seven hundred eighty-one patients were admitted from June 1994 to May 1999. A pelvic fracture was diagnosed in 660 (8.5%). Two hundred ninety (44.0%) dynamic helical CT scans were performed, of which 13 (4.5%) identified contrast extravasation. Nine (69%) were hemodynamically unstable and had pelvic arteriography performed. Arterial bleeding was confirmed in all and controlled by embolization. Patients with contrast extravasation had significantly greater Injury Severity Score, blood transfusion requirement and length of stay. Sensitivity, specificity, and accuracy of CT scan for identifying patients requiring embolization were 90.0%, 98.6%, and 98.3%, respectively. CONCLUSIONS: Early use of dynamic helical CT scanning in the multiply injured patient with a pelvic fracture accurately identifies the need for emergent angiographic embolization. PMID- 11015103 TI - Hepatic resection: effective treatment for primary and secondary tumors. AB - BACKGROUND: Hepatic resection is an accepted therapeutic modality for isolated colorectal metastases (CRM) and primary hepatobiliary cancers (PC). Controversy continues regarding the safety, efficacy, and appropriateness of resection for noncolorectal metastases (NCM). METHODS: A retrospective review of 167 resections in 160 patients was performed to evaluate the impact of demographics and perioperative data on survival and recurrence. Statistical analyses were performed by Student t test, analysis of variance, and Kaplan-Meier survival estimates. RESULTS: Resections were performed for CRM, 110 of 167 (66%), NCM, 31 of 167 (19%), and PC, 26 of 167 (15%). The interval from primary to metastases was significantly longer in the NCM group than the CRM group (34.7+/-45.1 vs. 18.7+/-23.7 months; P<.01). Mean number of lesions was not different between groups; however, NCM were larger than CRM (5.9+/-4.5 vs 4.5+/-2.9 cm; P<.05). Operative complications were significantly greater for PC (54%) versus CRM and NCM (21% and 19%, respectively; P<.01), although length of stay was similar between groups. Perioperative mortality was 2%. Actuarial survival at 1 year, 3 years, and 5 years was CRM 91%, 54%, and 40%, PC 75%, 60%, and 38%, and NCM 68%, 36%, and not available, respectively (CRM vs. NCM; P<.01 at 3 years). CONCLUSIONS: Hepatic resection for primary and secondary malignancy can be performed with minimal morbidity and mortality. Resection of NCM is associated with a lower overall survival compared with CRM and PC. The disease-free interval from resection of the primary to metastasectomy is prolonged and hepatic recurrence infrequent after resection in the NCM group. These results suggest that tumor biology is a critical determinant of outcome after hepatic resection of primary and secondary hepatic tumors. PMID- 11015104 TI - Safety and long-term efficacy of transduodenal excision for tumors of the ampulla of Vater. AB - BACKGROUND: Ampullary tumors should be resected because of the high incidence of malignancy and the unreliability of preoperative endoscopic diagnosis. Controversy exists about whether to perform a transduodenal excision (TDE) or a pancreatoduodenectomy. This study evaluated the safety and long-term efficacy of TDE. METHODS: The records of 21 patients with a pathologic diagnosis of ampullary adenoma who underwent TDE were reviewed. Demographics, symptoms, pathologic findings, and outcomes were analyzed and long-term follow-up was ascertained. RESULTS: Twenty-one patients (mean age, 61 years) underwent TDE. Final pathology showed adenoma in all patients including 1 (5%) with invasive cancer, 2 (9%) with microinvasive cancer, 6 (28%) with high-grade dysplasia, and 1 (5%) with low grade dysplasia. The overall survival was 85% (mean follow-up of 38 months). One of 3 late deaths was likely related to disease progression. Sixteen of the 18 remaining patients (89%) had no evidence of tumor recurrence. One benign ampullary recurrence was successfully treated endoscopically. One additional patient developed an ampullary cancer and underwent pancreatoduodenectomy. CONCLUSIONS: TDE of benign ampullary tumors, even those with varying grades of dysplasia, can be performed with acceptable morbidity and low rates of recurrence. Postoperative endoscopic surveillance is mandatory to identify recurrent tumors. PMID- 11015105 TI - Bilateral synchronous breast cancer: mode of detection and comparison of histologic features between the 2 breasts. AB - BACKGROUND: Bilateral synchronous breast cancer is uncommon (accounting for 1.0% 2.6% of all patients with breast cancer), and most physicians do not accumulate a large personal experience of patients with this disease. We reviewed our experience with patients with bilateral synchronous breast cancer, focusing on the mode of detection and histologic features in the 2 breasts. METHODS: The charts of patients who were treated at this institution for bilateral synchronous breast cancer during the 15-year period of 1984 through 1999 were reviewed. Information regarding age, mode of detection, histopathologic features, treatment, and overall survival were analyzed. RESULTS: During the study period, 51 patients (all women) were treated at our institution for bilateral synchronous breast cancer. This comprised 2.1% of all patients (n = 2382 patients) treated for breast cancer during the same period of time. The first cancer was detected by palpation in 81% and by mammography in 14%. The corresponding figures for the contralateral cancer were 24% and 54%, respectively. The histologic type of cancer was identical in the 2 breasts in 29 patients (57%) and was different between the 2 breasts in 22 patients (43%). The overall 10-year survival rate was 63%. CONCLUSIONS: Bilateral synchronous breast cancer is often detected by mammography and is frequently of the same histologic type as the index cancer. A better awareness of the risk for this disease may help detect bilateral breast cancer earlier. PMID- 11015106 TI - Prone positioning for acute respiratory distress syndrome in the surgical intensive care unit: who, when, and how long? AB - BACKGROUND: We evaluated the effects of prone positioning (PP) on surgery and trauma patients with acute respiratory distress syndrome (ARDS). METHODS: Patients with ARDS were studied. Exclusion criteria were contraindications to PP. Patients were evaluated in the supine position and after being turned to the PP. After 6 hours, patients were returned to the supine position for 3 hours. One hour after each position change, arterial and mixed venous blood was drawn and analyzed for blood gases and pH, and hemodynamics were measured. RESULTS: Over 20 months, 27 patients met the criteria, and 20 of the patients were entered into the study. On day 1, 18 of 20 patients (90%) responded with an increase in PaO(2) during PP. On day 2, 16 of 17 patients (94%) responded; on day 3, 15 of 16 patients responded (94%); on day 4, 11 of 13 patients responded (85%); on day 5, 8 of 8 patients responded (100%); and on day 6, 4 of 5 patients responded (80%). Pao(2)/Fio(2) and Qs/Qt were significantly improved (P<.05) during PP. There were 91 periods of PP, lasting 10.3+/-1.2 hours. Of 91 changes to PP, 78 changes (86%) resulted in an improvement in Pao(2)/Fio(2) of more than 20%. CONCLUSIONS: PP improves oxygenation in ARDS for 6 days with few complications. PMID- 11015107 TI - Angioplasty does not affect subsequent operative renal artery revascularization. AB - BACKGROUND: Although increased application of percutaneous renal artery angioplasty and stenting has facilitated nonoperative renal revascularization, patient outcomes after failed angioplasty are not established. METHODS: Renal artery revascularization was performed in 31 patients (38 arteries) from 1993 to 1999. Twenty patients underwent primary surgical repair, and 11 patients underwent secondary reconstruction after angioplasty (n = 7) or angioplasty and stenting (n = 4). Before operation, all patients had severe hypertension (blood pressure 166+/-5.2/92 +/- 2.7 mm Hg) that required an average of 3.0 +/- 0.2 medications for control. In addition, 12 patients (primary 45% vs secondary 27%; P = NS) had evidence of renal insufficiency (creatinine > or =1.7 mg/dL). RESULTS: There was no difference between primary and secondary procedures in the length of hospital stay (12+/- 1.4 vs. 12+/-3.2 days; P = NS), major morbidity (10% vs. 18%; P = NS) or perioperative mortality (overall mortality 2 of 31; primary 5% vs secondary 9%; P = NS). The majority of patients demonstrated improvement or cure of hypertension (primary 94% vs secondary 90%; P = NS) and stable or decreased creatinine (primary 74% vs secondary 82%; P = not significant). Overall survival (mean follow-up 22+/-3.5 months) was 89%+/-5.7%. CONCLUSIONS: Although this surgical series does not address the true outcomes of renal artery angioplasty, the results suggest that renal artery angioplasty does not prejudice subsequent surgical outcomes in patients who are carefully followed after angioplasty. PMID- 11015108 TI - Simultaneous pancreas-kidney transplantation in the mycophenolate mofetil/tacrolimus era: evolution from induction therapy with bladder drainage to noninduction therapy with enteric drainage. AB - BACKGROUND: In the past, enteric drainage or the omission of induction immunotherapy has been shown to be predictive of suboptimal outcomes of simultaneous pancreas-kidney (SPK) transplantation. We have reassessed the need for bladder drainage and induction immunotherapy to optimize the outcome of SPK transplantation. METHODS: One hundred consecutive recipients of SPK transplants who received mycophenolate mofetil and tacrolimus immunosuppression were studied. The first 50 recipients had bladder-drained pancreas allografts and received induction immunotherapy. The results were compared with the next 50 recipients who had enteric-drained pancreas allografts, which included a subgroup (n = 17 patients) who were randomized to receive no induction immunotherapy. RESULTS: The 1-year actuarial patient, kidney, and pancreas survival rates in the bladder drainage group were 98.0%, 94.0%, and 94.0%, respectively. The 1-year actuarial patient, kidney, and pancreas survival rates in the enteric-drainage group were 96.8%, 96.8%, and 89.4%, respectively. In the enteric-drainage group, the incidence of rejection at 1 year was 6.1% in recipients who received induction therapy versus 23.5% in recipients who did not receive induction therapy. The average number of readmissions per recipient was 1.8 in the bladder-drainage group versus 0.9 in the enteric-drainage group. CONCLUSIONS: Primary enteric drainage of the pancreas allograft in recipients of SPK transplantation is the preferred surgical technique in the tacrolimus/mycophenolate mofetil era. PMID- 11015109 TI - Mild ductal atypia after large-core needle biopsy of the breast: is surgical excision always necessary? AB - BACKGROUND: The aim of the current study was to identify a select group of patients with mild atypia who do not need surgical excision after large-core needle biopsy (LCNB) of the breast. METHODS: Nineteen (70%) of 27 patients with ductal atypia found on LCNB had subsequent surgical excision. These 19 patients were retrospectively assigned to 3 groups according to the severity of the atypia found, which was compared with the final pathologic specimen after surgical biopsy. RESULTS: Cancer was identified through surgical biopsy in 6 (32%) of 19 patients. The severity of atypia seen on the LCNB specimen strongly correlated with subsequent cancer identification (P<.01). Two (33%) of 6 patients in group 2 (true atypical ductal hyperplasia [ADH]) and 4 (80%) of 5 patients in group 3 (severe ADH, borderline ductal carcinoma in situ) had cancer after surgical biopsy. No cancer was found after surgical biopsy in 8 patients in group 1 (mild atypia, not meeting criteria for ADH). CONCLUSIONS: The results of this study suggest that surgical excision can be avoided after LCNB of the breast in patients with only mildly atypical lesions that do not meet criteria for ADH. Patients with true ADH should continue to have surgical excision. PMID- 11015110 TI - Intraoperative sestamibi scanning in reoperative parathyroidectomy. AB - BACKGROUND: Reoperative neck exploration for hyperparathyroidism is often difficult even for experienced surgeons. Recent advances in preoperative and intraoperative localization techniques have improved successful resection rates. This prospective study evaluates the accuracy and clinical utility of intraoperative technetium 99m sestamibi scanning for localizing hyperfunctioning parathyroid tissue in reoperative neck explorations. PATIENTS AND METHODS: Eleven patients underwent reoperative neck exploration for hyperparathyroidism. Two patients had 3 prior neck explorations, 1 had 2 prior neck explorations, and 8 patients had 1 prior neck operation. Preoperative studies included sestamibi scintigraphy and ultrasound in all patients, magnetic resonance imaging in 4, computed tomography scan in 3, parathyroid arteriogram in 1, and selective venous sampling in 1. All patients underwent intraoperative technetium 99m sestamibi scanning and parathyroid hormone assay. RESULTS: Preoperative technetium 99m sestamibi scanning and ultrasound each successfully localized 7 of 11 hyperfunctioning glands (64%). Intraoperative technetium 99m sestamibi scanning correctly localized 10 of 11 hyperfunctioning glands (91%). Intraoperative parathyroid hormone assay confirmed successful excision of hyperfunctioning tissue in all 11 patients. Postoperatively, all 11 patients had low-normal or normal calcium levels. CONCLUSIONS: Intraoperative technetium 99m sestamibi correctly localized 91% of hyperfunctioning glands compared with 64% localization for preoperative technetium 99m sestamibi and preoperative ultrasound. Intraoperative technetium 99m sestamibi scanning and parathyroid hormone monitoring are useful in reoperative neck explorations for hyperparathyroidism. PMID- 11015111 TI - Less invasive aortic surgery: the minilaparotomy technique. AB - BACKGROUND: This study evaluated a less invasive technique for exposure of the infrarenal aorta and its impact on the treatment of patients with abdominal aortic aneurysms (AAA) or aortoiliac occlusive disease (AIOD), or both. METHODS: Forty patients with AAA (26), aneurysmal extension into the iliac arteries (6), or AIOD (8) were prospectively selected for minilaparotomy aortic exposure and repair using a small periumbilical midline incision (< or =10 cm); intra abdominal, nondisplaced retraction of the small bowel; and conventional hand-sewn vascular anastomoses. Perioperative comparisons with a contemporary group of AAA patients treated with long, open midline incision and extracavitary small bowel retraction were made. RESULTS: There was no significant difference between the minilaparotomy and open surgical control groups for operating room time; intraoperative, perioperative morbidity; or mortality. Significant differences were documented between the minilaparotomy and the control group with regard to stay in the intensive care unit (days; 1.0+/-1.2 versus 1.8+/-1.5); return to general diet (days; 3+/-1.3 versus 4.7+/-2.8); and length of stay (days; 4.9+/ 1.8 versus 7.3+/- 3.4). CONCLUSIONS: Minilaparotomy exposure is safe and effective for treatment of infrarenal AAA and AIOD. This technique maintains quality outcome while reducing postoperative ileus, hospital stay, and resource utilization. PMID- 11015112 TI - Effects of an increased air gap on the in vitro interaction of wireless phones with cardiac pacemakers. AB - Several clinical and laboratory studies have demonstrated electromagnetic interaction between implantable cardiac pacemakers and hand-held wireless phones operated in close proximity. Current FDA and HIMA labeling guidelines indicate that a minimum separation of 6 in (15 cm) should be maintained between a hand held wireless phone and an implanted pacemaker. This separation requirement does not distinguish between lateral locations on the chest and a perpendicular air gap. Evidence is provided here for a substantially reduced separation threshold when measured across an air gap rather than near the saline conductive media of a simulated torso. Twenty pacemaker-phone combinations involving 6 pacemakers and 9 phones were evaluated in vitro under worst-case conditions with respect to phone output power and pacemaker sensitivity. The phones represented CDMA, TDMA-11 Hz, TDMA-22 Hz, TDMA-50 Hz, and TDMA-217 Hz digital wireless technologies. Small increases in the perpendicular air gap between the phone and the saline surface resulted in a dramatic reduction in interaction. Approximately half of the 208 test runs exhibiting interaction at an air gap of 1 cm no longer resulted in interaction when the gap was increased to 2 cm. At a gap of 7.4 cm, the percentage of runs with interaction decreased to 1.4%. The overall interaction rate, considering a total of 8296 test runs from an earlier study, was less than 0.07% at a total perpendicular distance of 8.6 cm from the saline surface to the phone antenna axis. The perpendicular distance threshold of 8.6 cm was significantly less than the horizontal plane projection threshold of 19 cm previously reported. This difference is a function of the electromagnetic field coupling to the saline bath rather than field strength changes along the axis of the phone antenna. The results have implications for those making recommendations to pacemaker patients who may be unaware of this distinction. PMID- 11015113 TI - Effect of low frequency, low amplitude magnetic fields on the permeability of cationic liposomes entrapping carbonic anhydrase: I. Evidence for charged lipid involvement. AB - The influence of low frequency (4-16 Hz), low amplitude (25-75 mu T) magnetic fields on the diffusion processes in enzyme-loaded unilamellar liposomes as bioreactors was studied. Cationic liposomes containing dipalmitoylphosphatidylcholine, cholesterol, and charged lipid stearylamine (SA) at different molar ratios (6:3:1 or 5:3:2) were used. Previous kinetic experiments showed a very low self-diffusion rate of the substrate p-nitrophenyl acetate (p-NPA) across intact liposome bilayer. After 60 min of exposure to 7 Hz sinusoidal (50 mu T peak) and parallel static (50 mu T) magnetic fields the enzyme activity, as a function of increased diffusion rate of p-NPA, rose from 17 +/- 3% to 80 +/- 9% (P < .0005, n = 15) in the 5:3:2 liposomes. This effect was dependent on the SA concentration in the liposomes. Only the presence of combined sinusoidal (AC) and static (DC) magnetic fields affected the p-NPA diffusion rates. No enzyme leakage was observed. Such studies suggest a plausible link between the action of extremely low frequency magnetic field on charged lipids and a change of membrane permeability. PMID- 11015114 TI - Effect of low frequency, low amplitude magnetic fields on the permeability of cationic liposomes entrapping carbonic anhydrase: II. No evidence for surface enzyme involvement. AB - Observations recently reported by our group indicate that combined 7 Hz sinusoidal (B(acpeak) = 50 mu T) and parallel static (B(dc) = 50 mu T) magnetic fields can induce a significant increase in diffusion rate of substrate across carbonic anhydrase (CA)-loaded liposomes (DPPC:Chol:SA). A direct involvement of charges of stearylamine (SA) on the lipid membrane surface was also demonstrated. Kinetic studies showed that CA was mainly entrapped in liposomes at 5:3:2 molar ratio, although a small amount (17%) of enzyme was also located on the external surface of these cationic liposomes. In this paper we report steady state kinetic studies on this latter CA after ELF-EMFs exposure. No difference in the apparent K(m) between exposed and sham samples was observed. On the contrary the apparent V(max) was increased by approximately a factor of 2 after field exposure. In spite of the proteolytic digestion of this external CA, a significant increase of enzymatic activity, as a function of increase in the diffusion rate of substrate across the lipid bilayer, was observed in the exposed samples. Based on these results, a conformational change induced by the field on the CA located on the external surface of 5:3:2 liposomes is excluded as an explanation for our previous observations, supporting the primary role of bilayer SA in the interaction with ELF. A model of ELF interaction, based on the Larmor precession theory, explaining the physical phenomenon induced on the dipole of SA has been developed. PMID- 11015115 TI - Recommended minimal requirements and development guidelines for exposure setups of bio-experiments addressing the health risk concern of wireless communications. AB - The evidence currently available on the potential health effects from electromagnetic field (EMF) exposure has been largely judged as being too tentative and inadequate to meet criteria for assessing health risks. Some of the main reasons for these shortcomings lie in the incomplete description of the exposure and poorly characterized dosimetry. Well-defined exposure conditions are essential to obtain reproducible and scientifically valuable results. To facilitate the development of optimized setups for specific bio-experiments, this paper lists basic requirements and provides development guidelines for evaluation, optimization, construction, and verification of exposure. In addition, definitions of minimum performance requirements for setups addressing the health risk concern of wireless communications are suggested. PMID- 11015116 TI - No mutagenic or recombinogenic effects of mobile phone fields at 900 MHz detected in the yeast Saccharomyces cerevisiae. AB - Both actively growing and resting cells of the yeast Saccharomyces cerevisiae were exposed to 900-MHz fields that closely matched the Global System for Mobile Communication (GSM) pulsed modulation format signals for mobile phones at specific absorption rates (SAR) of 0.13 and 1.3 W/kg. Two identical anechoic test chambers were constructed to perform concurrent control and test experiments under well-controlled exposure conditions. Using specific test strains, we examined the genotoxic potential of mobile phone fields, alone and in combination, with a known genotoxic compound, the alkylating agent methyl methansulfonate. Mutation rates were monitored by two test systems, a widely used gene-specific forward mutation assay at CAN1 and a wide-range assay measuring the induction of respiration-deficient (petite) clones that have lost their mitochondrial function. In addition, two further assays measured the recombinogenic effect of mobile phone fields to detect possible effects on genomic stability: First, an intrachromosomal, deletion-formation assay previously developed for genotoxic screening; and second, an intragenic recombination assay in the ADE2 gene. Fluctuation tests failed to detect any significant effect of mobile phone fields on forward mutation rates at CAN1, on the frequency of petite formation, on rates of intrachromosomal deletion formation, or on rates of intragenic recombination in the absence or presence of the genotoxic agent methyl methansulfonate. PMID- 11015117 TI - Neural and behavioral teratological evaluation of rats exposed to ultra-wideband electromagnetic fields. AB - Several investigators have reported teratologic effects of electromagnetic field exposure. The majority of these studies have been performed at levels of exposure that could produce substantial heating of the animals. New and unique sources of ultra-wideband (UWB) electromagnetic fields are currently being developed and tested that are capable of generating nonthermalizing, high-peak-power, microwave (MW) pulses with nanosecond (ns) pulse widths, picosecond (ps) rise times, and an UWB of frequencies. Our study was performed to determine if teratological changes occur in rat pups as a result of (i) daily UWB exposures during gestation days 3 18, or (ii) as a result of both prenatal and postnatal (10 days) exposures. Dams were exposed either to (i) UWB irradiation from a Kentech system that emitted a 55 kV/m-peak E field, 300 ps rise time, and a 1.8 ns pulse width, average whole body specific absorption rate 45 mW/kg; (ii) sham irradiation; or (iii) a positive control, lead (Pb) acetate solution (2000 microg/ml) continuously available in the drinking water. Offspring were examined for ontogeny (litter size, sex-ratios, weights, coat appearance, tooth-eruption, eye-opening, air righting, and ultrasonic stress vocalizations). Male pups were tested on various performance measures (locomotor, water-maze learning, and fertilization capabilities). The pups postnatally exposed were examined for hippocampal morphology and operant behavior. Behavioral, functional, and morphological effects of UWB exposure were unremarkable with these exceptions: (i) The UWB exposed pups emitted significantly more stress vocalizations than the sham exposed pups; (ii) the medial-to-lateral length of the hippocampus was significantly longer in the UWB-exposed pups than in the sham-exposed animals; (iii) male offspring exposed in utero to UWB mated significantly less frequently than sham-exposed males, but when they did mate there was no difference in fertilization and offspring numbers from the sham group. There does not appear to be a unifying physiological or behavioral relationship among the significant differences observed, and our findings could be due to the expected spurious results derived when a large number of statistical comparisons are made. Significant effects found between our positive-controls and other groups on numerous measures indicates that the techniques used were sensitive enough to detect teratological effects. Bioelectromagnetics 21:524-537, 2000. Published 2000 Wiley-Liss, Inc. PMID- 11015118 TI - The possible role of contact current in cancer risk associated with residential magnetic fields. AB - Residential electrical wiring safety practices in the US result in the possibility of a small voltage (up to a few tenths of a volt) on appliance surfaces with respect to water pipes or other grounded surfaces. This "open circuit voltage" (V(OC)) will cause "contact current" to flow in a person who touches the appliance and completes an electrical circuit to ground. This paper presents data suggesting that contact current due to V(OC) is an exposure that may explain the reported associations of residential magnetic fields with childhood leukemia. Our analysis is based on a computer model of a 40 house (single-unit, detached dwelling) neighborhood with electrical service that is representative of US grounding practices. The analysis was motivated by recent research suggesting that the physical location of power lines in the backyard, in contrast to the street, may be relevant to a relationship of power lines with childhood leukemia. In the model, the highest magnetic field levels and V(OC)s were both associated with backyard lines, and the highest V(OC)s were also associated with long ground paths in the residence. Across the entire neighborhood, magnetic field exposure was highly correlated with V(OC) (r = 0.93). Dosimetric modeling indicates that, compared to a very high residential level of a uniform horizontal magnetic field (10 mu T) or a vertical electric field (100 V/m), a modest level of contact current (approximately 18 mu A) leads to considerably greater induced electric fields (> 1 mV/m) averaged across tissue, such as bone marrow and heart. The correlation of V(OC) with magnetic fields in the model, combined with the dose estimates, lead us to conclude that V(OC) is a potentially important exposure with respect to childhood leukemia risks associated with residential magnetic fields. These findings, nonetheless, may not apply to residential service used in several European countries or to the Scandinavian studies concerned with populations exposed to magnetic fields from overhead transmission lines. PMID- 11015119 TI - Vitellogenin of Blattella germanica (L.) (Dictyoptera, blattellidae): nucleotide sequence of the cDNA and analysis of the protein primary structure. AB - The cloning and sequencing of a cDNA of the vitellogenin gene from the cockroach Blattella germanica is reported. It is 5,749 nucleotides long and encodes an amino acid sequence of 1,862 residues (including a putative signal peptide of 17 residues). The vitellogenin sequence includes a long serine-rich stretch between amino acids 322 and 349, and two other stretches between amino acids 1691 and 1740. The vitellogenin of B. germanica shows a notable similarity (between 32 and 42%) to those described in other insects, and its alignment shows a high number of motifs conserved in all species, especially in the subdomains I-V. Non parsimony methods (Neighbor Joining) of phylogenetic analysis of the insect vitellogenin sequences gave a tree showing a topology that is, in general, congruent with the currently accepted insect phylogenetic schemes. Arch. PMID- 11015120 TI - Apoptosis in cultured midgut cells from heliothis virescens larvae exposed to various conditions. AB - We exposed midgut cells from primary cultures of Heliothis virescens larvae to cell-free previously used medium, the Vaughn X and HyQ SFtrade mark media used for serum-free culture of insect cell lines which do not support H. virescens midgut cells, and to toxin from Bacillus thuringiensis. A statistically significant increase in the percent of dying cells was counted in cell populations in Vaughn X medium. Use of the TUNEL method to detect apoptosis indicated a low rate (7.2%) of apoptosis in control cultures grown in Heliothis medium, an increase to approximately 20% in previously used and HyQ SFtrade mark media, and to approximately 45% of cells remaining after exposure to and initial destruction by B. thuringiensis toxin. Apoptotic nuclei were predominant (approximately 6%) in mature columnar cells in control cultures. Approximately 1% of goblet, stem, and differentiating cells were apoptotic. However, apoptosis rose to 12% in stem and differentiating cells exposed to used and unsuitable medium. B. thuringiensis exposure to toxin for 2-3 days resulted in visible membrane damage and necrosis, causing the death of 84% of the cells as measured by both the TUNEL and Annexin methods. Some of the columnar cells and stem and differentiating cells that remained also contained apoptotic nuclei. Stem and differentiating cells normally replace dying mature cells in the midgut. Thus, exposure of cultures of H. virescens midgut cells to adverse environments such as unsuitable or poisonous media appeared to induce down-regulation of the cell populations by apoptosis. PMID- 11015121 TI - A cDNA encoding a chitinase from the epithelial cell line of chironomus tentans (Insecta, diptera) and its functional expression. AB - A cDNA coding for chitinase was isolated from Chironomus cells, which possesses conserved regions I and II characteristic for family 18 chitinases, a C-terminus enriched in Glu and Pro without the typical "PEST-region," putative glycosylation sites, a reduced number of C-terminal cysteines, and no typical chitin binding domain. Northern blots revealed one specific signal with an apparent size of 2.3 kb. The cDNA was expressed in the baculovirus/Spodoptera system as a His-tag fusion protein, which was secreted as a functionally active enzyme into the medium and could be separated from endogenous viral and Spodoptera-specific chitinases. PMID- 11015122 TI - Cloning of vitellogenin cDNA of the American cockroach, Periplaneta americana (Dictyoptera), and its structural and expression analyses. AB - A cDNA expression library constructed from poly (A)(+) RNA prepared from vitellogenic female fat body cells of the American cockroach, Periplaneta americana (Dictyoptera) was screened using a polyclonal antiserum against the 100 kD polypeptide(s) from the egg extract. A partial Vg cDNA clone was obtained and sequenced. The 5' end portion of the cDNA was then obtained by the RACE method, cloned, and sequenced. The combined complete Vg cDNA was 5,854 bp long and contained a single ORF encoding 1,896 amino acids. The entire deduced amino acid sequence was aligned confidently with those of the known insect Vgs. A GL/ICG motif, a number of cysteines at conserved locations following this motif, and a DGXR motif upstream of the GL/ICG motif were present near the C-terminal. The chemically determined N-terminal amino acid sequence of the 170-kD polypeptide from the egg extract completely matched the deduced sequence starting from just after one of the consensus (RXXR) cleavage sites, indicating the occurrence of post-translational cleavage in the fat body cells. The Vg gene begins to be expressed in the 2-day-old adult female fat body cells but is never expressed in ovaries or in male fat body cells. Hemolymph Vg was first detected by immunoblotting in 4-day-old adult females, 2 days after the beginning of gene expression. Western blot analysis of major yolk polypeptides in nine cockroach species belonging to the two superfamilies, Blattoidea and Blaberoidea, using the antisera against P. americana major yolk polypeptides showed that the similarities in Vn antigenicity are basically limited to within a superfamily. PMID- 11015123 TI - Relationship between case-control studies and the transmission/disequilibrium test. AB - Case-control studies provide a powerful approach for detecting disease susceptibility loci that have only a weak to moderate impact on the risk of disease, or markers that are in linkage disequilibrium with such loci. However, since any association detected in a case-control study may result from uncontrolled confounding, evidence for disease-marker associations obtained from such studies must be confirmed by alternative methods. Since studies that use the transmission/disequilibrium test or TDT are frequently employed to confirm disease-marker associations detected in case-control studies, data are increasingly available from both case-control studies and "TDT studies" of the same disease-marker association. It would, therefore, be useful to have a single measure of the magnitude of the disease-marker association that would allow for comparison of results from these two study designs. Such a measure could also be used to estimate minimum sample size requirements for TDT studies of previously reported disease-marker associations. An obvious measure of the disease-marker association in TDT studies is the frequency (T) with which heterozygous parents transmit the putative, high-risk marker allele to affected offspring. In this paper, it is shown that T can also be estimated from case-control data with a minimum of assumptions, and that T is the critical parameter for determining power and estimating sample sizes for the TDT. PMID- 11015124 TI - Estimation of apolipoprotein E genotype-specific relative mortality risks from the distribution of genotypes in centenarians and middle-aged men: apolipoprotein E gene is a "frailty gene," not a "longevity gene". AB - We developed a method to estimate genotype-specific average relative mortality risk, R, from genotype distributions in cross-sectional studies of people belonging to different age-groups, and applied the method to new data from a study of apolipoprotein E genotypes (apoE) in 177 Danish centenarians and data from a study of 40-year-old Danish men. Twenty-one percent of the centenarians were epsilon 2-carriers (genotypes epsilon 2 epsilon 2 and epsilon 3 epsilon 2) and 15% were epsilon 4-carriers (genotypes epsilon 4 epsilon 4 and epsilon 4 epsilon 3) compared to 13 and 29%, respectively, of the young men. The R-values were 0.95 (95% CI 0.88 to 1.02) for epsilon 2-carriers and 1.13 (95% CI 1.05 to 1.22) for epsilon 4-carriers, using epsilon 3 epsilon 3- and epsilon 4 epsilon 2 genotypes as reference. Corresponding values for epsilon 4-carriers were obtained by using published data from a French and a Finnish study of centenarians, whereas the values for epsilon 2-carriers were about 0.90 with these data. The method to estimate mortality risk and the results associate with the view that the apoE gene is a "frailty gene." On the other hand, if odds ratios are used to summarize data from studies of this kind, they are more impressive and may propagate the misconception that apoE is a "longevity gene". PMID- 11015125 TI - Combined linkage and linkage disequilibrium mapping for genome screens. AB - Linkage analysis and association studies, two major approaches for genetic studies of human diseases, are useful for mapping genes that are highly penetrant, but both use only part of the information that is available for mapping disease genes. Therefore, they provide limited utility when used alone. In this report, we present combined linkage and linkage disequilibrium mapping that simultaneously utilizes linkage and linkage disequilibrium information for mapping human disease genes. Compared with the existing linkage analysis and association study methods, this method has several advantages: 1) it has high statistical power by a joint analysis of linkage and linkage disequilibrium for localizing disease susceptibility loci: 2) it unifies the theory of linkage analysis and linkage disequilibrium mapping, 3) it retains the general framework for linkage analysis and, hence, can be easily incorporated into the existing software for the linkage analysis. The proposed LLDM is applied to familial hemophagocytic lymphohistiocytosis (FHL) disease. PMID- 11015126 TI - Effects of early inhaled beclomethasone therapy on tracheal aspirate inflammatory mediators IL-8 and IL-1ra in ventilated preterm infants at risk for bronchopulmonary dysplasia. AB - We tested the hypothesis that inhaled beclomethasone therapy for prevention of bronchopulmonary dysplasia (BPD) reduces pulmonary inflammation. As part of a randomized, placebo-controlled trial, interleukin-8 (IL-8) and interleukin-1 receptor antagonist (IL-1ra) concentrations in tracheal aspirates were measured as markers of pulmonary inflammation. On study days 1 (baseline), 8, 15, and day 28 of age, samples were obtained from enrolled infants (birth weights <1,251 g, gestational age <33 week, 3 to 14 days of age) who remained ventilated and had not received systemic glucocorticoid therapy. Cytokine levels (pg/microg of free secretory component of immunoglobulin A) were compared between groups. We determined whether baseline cytokine levels modified treatment effect regarding subsequent need for systemic glucocorticoid therapy or occurrence of BPD (age 28 days). Tracheal aspirates were obtained from 161 infants (77 receiving beclomethasone, 84 receiving placebo). Median IL-8 levels were lower in beclomethasone versus placebo infants on study days 8 (82.9 vs. 209.2, P < 0.01) and 15 (37.4 vs. 77.4, P < 0.03) after controlling for antenatal glucocorticoid therapy and maternal race. Median IL-1ra levels were lower in beclomethasone versus placebo infants only on study day 8 (86.5 vs. 153.3, P < 0.01). Fewer beclomethasone infants with baseline IL-8 levels in the interquartile range required systemic glucocorticoid therapy (beclomethasone 30.6% vs. placebo 65.8%, P < 0.01) or developed BPD (beclomethasone 42.4% vs. placebo 69.4%, P < 0.03). We conclude that early-inhaled beclomethasone therapy was associated with a reduction in pulmonary inflammation after 1 week of therapy. Beclomethasone treated infants with moderately elevated baseline IL-8 levels received less subsequent systemic glucocorticoid therapy and had a lower incidence of BPD than nontreated infants. PMID- 11015127 TI - Early changes in respiratory compliance and resistance during the development of bronchopulmonary dysplasia in the era of surfactant therapy. AB - Despite the availability of surfactant treatment, extremely low birth weight (ELBW) infants continue to be at high risk of developing bronchopulmonray dysplasia (BPD). Evidence suggests that pathologic changes occur within the first few days of life. We hypothesized that the changes in early respiratory system compliance and resistance in ELBW infants with or without hyaline membrane disease (HMD) would correlate with BPD severity and aid in its prediction. Respiratory system compliance (Crs) and resistance (Rrs) were measured at the end of weeks 1, 2, 3, and 4 in 46 infants weighing 1,000 g or less at birth, using the single breath airway occlusion method. Twenty-four infants had HMD and 22 did not. Fifteen infants with and 10 infants without HMD developed BPD with radiological changes and oxygen needs at 28 days. Twelve BPD infants required supplemental oxygen beyond 36 weeks, defined as chronic lung disease (CLD). Irrespective of whether the infant initially had HMD, the week 1 results showed that infants who subsequently developed BPD had a significantly higher respiratory system resistance than those who did not (P = 0.0014). Though week 1 compliance was lower, it was not statistical significant. Multiple logistic models consisting of simple neonatal variables and week 1 respiratory mechanics showed that Rrs was independently associated with subsequent BPD (P = 0.026) and CLD (P = 0.016), while compliance was not. Prediction of CLD improved with the inclusion of Rrs results as compared to prediction using clinical variables alone. Throughout the 4-week study period, Rrs was significantly higher in BPD infants than in those without BPD, and resistance was particularly abnormal in those who had CLD or subsequently required corticosteroid treatment. These observations provide rationale for interventions to prevent BPD within the first week of life. Respiratory mechanics measurements could be useful in the assessment of therapeutics in the current surfactant era. PMID- 11015128 TI - Very low birthweight and asthma by age seven years in a national cohort. AB - Several studies have suggested that very low birthweight (VLBW < 1500 g) is associated with increased rates of respiratory problems in childhood and that the presence of chronic lung disease further increases the risk. We aimed to assess rates of asthma at 7-8 years of age in a national cohort of VLBW infants born in 1986 and for whom perinatal data were available. Two hundred ninety-nine former VLBW children (96% of surviving children living in New Zealand) were assessed at a home visit. Parents were asked a comprehensive questionnaire, including three questions aimed at assessing morbidity from asthma: 1) was the child diagnosed as having asthma before age 7 years; 2) was the child still experiencing asthma at the age of 7 years; and 3) was the child prescribed daily medication for asthma at the age of 7 years. Overall, 50% of the cohort had been diagnosed with asthma before age 7, compared with 27% of a sample of New Zealand children assessed contemporaneously in an international study; 32% had asthma at age 7, and 11% were taking daily medication. All three categories of asthma were associated with a family history of asthma, but there was no association with any perinatal factors. A diagnosis of asthma before age 7 was more likely when the mother smoked in pregnancy (P < 0.005) and currently smoked (P < 0.01), and trended so when parents lacked high school qualifications and in Maori or Pacific Island families (P < 0.10). In contrast, daily medication was more frequent when parents had educational qualifications and in non-Maori or Pacific Island families (P < 0.05). On multiple logistic regression, a family history of asthma was a significant predictor for any and current asthma (P < 0.001) and daily medication (P < 0.05); maternal smoking in pregnancy was a significant predictor for any asthma (P < 0.05); and non-Maori or Pacific Island ethnicity was a significant predictor for asthma treatment (P < 0.05). We conclude that rates of childhood asthma are high in this VLBW cohort, but the high prevalence appears to be unrelated to perinatal factors, including respiratory morbidity. There are suggestions that social factors contribute to both asthma risk and treatment. PMID- 11015130 TI - Family history of atopy and clinical course of RSV infection in ambulatory and hospitalized infants. AB - Respiratory syncytial virus (RSV) infection can be severe in pediatric patients. Risk factors for severe disease include age less than 6 months, prematurity, preexisting heart or lung disease or malformations, gastroesophageal reflux, and immunodeficiency. The aim of the present study was to investigate the influence of family history of allergy on the clinical course of RSV infection in ambulatory and hospitalized infants. In a retrospective study, 172 patients younger than 12 months of age (99 inpatients and 73 outpatients) were enrolled. Information was obtained from hospital charts and from questionnaires sent to pediatricians. Inpatients had a significantly higher rate of atopy in their family history than outpatients, 62% and 29%, respectively (P < 0.001). Bronchiolitis was diagnosed more frequently in patients with an atopic burden than those without, 89% versus 74%, respectively (P < 0.02). Inpatients with an atopic family history had a significantly longer hospital stay than those without such a history, 7.4 +/- 3.7 days and 6.1 +/- 2.3 days, respectively (P < 0.04). Factors other than age that are considered a risk for severe infection with RSV (prematurity, preexisting heart or lung disease or malformation, and gastroesophageal reflux) were not confirmed in the present study. We conclude that infants with a family history of atopy are at increased risk for severe RSV infection as indicated by higher rates of hospitalization, longer hospital stay, and more frequent occurrence of bronchiolitis. PMID- 11015129 TI - School-based identification of asthma in a low-income population. AB - The increase in the prevalence, morbidity, and mortality of asthma among children over the last decade has been well documented, especially among low-income minority children. Hypotheses for the increases in morbidity and mortality include limited access to primary care services and the failure to recognize the presence and severity of asthma. The University of Miami Pediatric Mobile Clinic (Mobile Clinic) Asthma Intervention Program is designed to identify underserved asthmatic children at school and offer them culturally sensitive care. Nine elementary schools with low income, predominantly Hispanic and African-American populations regularly served by the Mobile Clinic, were chosen for study participation. All 5,800 students who were enrolled in kindergarten through third grade were given letters at the time of registration by their homeroom teachers about the asthma program. Caretakers who returned the questionnaire and reported that the student had asthma symptoms were invited to have the student undergo a medical evaluation in the Mobile Clinic. Over a 2-year period, caretakers of 423 students (7.3% of all students) expressed an interest in further evaluating their child's respiratory health. Of these, we enrolled and evaluated 154 in the Mobile Clinic's Asthma Intervention Program. The Mobile Clinic physicians identified 145 of the enrollees as having asthma. These results indicate that in elementary schools serving predominantly low-income minority populations, a large fraction of the asthmatic population (estimated prevalence, 6-10%) can be identified by a school-based letter. Further, in a subset of asthmatic students (children of interested caretakers), there is good agreement between caretaker responses and physician diagnosis of asthma. Since school attendance is mandatory, school-based methods may be an effective method for identifying low-income children with asthma. PMID- 11015131 TI - Limited agreement between written and video asthma symptom questionnaires. AB - The prevalence of asthma remains difficult to determine with precision with no absolute or "gold" standard for diagnosis. A recently developed video questionnaire for epidemiological studies with less reliance on understanding written questions provides another tool for determining prevalence and severity of asthma. This report from the International Study of Asthma and Allergies in Childhood (ISAAC) examines the agreement between the ISAAC video questionnaires on respiratory symptoms and reported asthma. Between December 1993 and April 1995, 4952 children aged 13-14 years in two Canadian communities completed sequentially the ISAAC written and video questionnaires at school. The agreement between responses to the two questionnaires for reported wheeze ever, current wheeze, wheeze on exercise, and nocturnal wheeze (the latter three questions relating to symptoms in the last 12 months), and to any combination of the latter three questions was examined in the full sample and in those reporting diagnosed asthma, using concordance and kappa coefficients as measures of agreement. The prevalences of wheeze ever, current wheeze, wheeze on exercise, and nocturnal wheeze were significantly lower based on responses to the video questionnaire compared with the written questionnaire in both regions in the full sample and in those labeled as having asthma. Although concordance between video and written questionnaires always exceeded 60% and often exceeded 70% for related questions, agreement measured by the kappa statistic for each question was only fair to moderate (kappa = 0.22-0.51). We conclude that the video questionnaire yields lower reported prevalence rates for asthma symptoms, and that there is limited agreement between responses to the two questionnaires that is not explained by issues of language, culture, or literacy. PMID- 11015132 TI - Bronchoalveolar lavage and esophageal pH monitoring data in children with "difficult to treat" respiratory symptoms. AB - Gastroesophageal reflux (GER) may be associated with chronic or recurrent asthma like symptoms secondary to bronchoconstrictor reflexes and/or inhalation of gastric content. The presence of lipid-laden alveolar macrophages has been proposed as an index to establish the degree of gastric aspiration. We evaluated 20 children with "difficult to treat" respiratory symptoms and a clinical history suggestive of GER. All children underwent 24-hr esophageal pH monitoring (pHm) and fiberoptic bronchoscopy with bronchoalveolar lavage (BAL). The amount of lipid per single macrophage was determined by a semiquantitative method, using fluorescence microscopy to detect Nile-Red-stained BAL cells and calculating a lipid-laden macrophage index (LLMI). Eleven children had positive pHm recordings, suggesting the presence of GER (pH-positive patients), and 9 had negative pHm records (pH-negative patients). The pH-positive patients had higher percentages of neutrophils and higher LLMI than the pH-negative children (P < 0.05). There were no correlations between the pHm records and either % BAL neutrophils or LLMI in pH-positive or pH-negative patients (P > 0.05; each correlation). In contrast, a single correlation was found between % BAL neuytrophils and LLMI, both in the pH-positive and in the pH-negative patients (r = 0.72, P = 0.02 and r = 0.71, P = 0.04, respectively). These data demonstrate that a significant proportion of pH positive patients with respiratory symptoms have BAL abnormalities that suggest airway inflammation and gastric content aspiration. However, the intensity of GER as indicated by pH monitoring does not correspond with BAL data in all patients. PMID- 11015133 TI - Serum lipase levels as a diagnostic marker in cystic fibrosis patients with normal or borderline sweat tests. AB - Patients with normal or borderline sweat test present a diagnostic challenge. In spite of the availability of different methods such as genetic analysis and measurements of nasal potential difference, uncertainty in diagnosing cystic fibrosis (CF) in some patients still exists. Neonates with CF have high serum lipase levels, which decline over time in pancreatic-insufficient patients, whereas pancreatic-sufficient patients demonstrate high serum lipase levels beyond infancy. Because patients with borderline or normal sweat test are almost always pancreatic sufficient, this study was aimed to assess whether serum lipase levels may be of help in establishing the diagnosis of CF in these patients. Serum lipase levels were measured in 100 CF patients and in 17 healthy individuals. Patients were grouped according to their genotype. Group A patients (n = 70) carried two mutations previously found to be associated with a pathologic sweat test and pancreatic insufficiency (delta F508, W1282X, G542X, N1303K, S549R). Group B (n = 30) were compound heterozygote patients who carried one mutation known to cause mild disease with borderline or normal sweat tests and pancreatic sufficiency (3849+10kb C-->T, 5T). Group C included 17 healthy controls. Serum lipase levels ranged between 2 and 104.4 U/L (mean +/- SD 16.9 +/ 14.7), 6.1-200 U/L (mean +/- SD 53.9 +/- 47.9), and 8.5-27.8 U/L (mean +/- SD 16.9 +/- 5.1) in Groups A, B, and C, respectively, with some overlapping between groups. The distribution of lipase levels was significantly different in Group B vs Groups A and C (P < 0.01). High lipase levels were found in 63.3% (19/30) of Group B patients, but in only 4.3% (3/70) and 0% (0/17) of Group A and C, respectively. Lipase levels were found to be inversely related to sweat chloride concentrations (r = -0.19, P < 0.05). Patients with borderline or normal sweat tests had high lipase levels, whereas low lipase levels were associated with pathologic sweat tests. Our findings indicate that the serum lipase level is genetically determined and that it has a useful role in the diagnosis of CF. Thus, in patients with borderline sweat tests and high lipase levels, the diagnosis of CF should be considered. PMID- 11015134 TI - Perfluorochemical elimination from the lungs: effect of initial dose. AB - Liquid-assisted ventilation with perfluorochemical (PFC) has been beneficial in a variety of respiratory diseases in animals and humans. Although PFC evaporation from the lungs is in part dependent on ventilation strategy and positioning, guidelines for initial and replacement dosing are unclear. We hypothesized that PFC evaporative loss over time is dependent on the size of the initial dose. Juvenile rabbits (n = 18) were ventilated using constant animal position and ventilator strategy. PFC (perflubron: LiquiVent ) was instilled endotracheally, using four groups with initial doses of 2, 6, 12, and 17 mL/kg. A previously described thermal detector that measures PFC in expired gas was used to calculate loss rate, residual perflubron in the lung, and volume loss as a % of initial fill volume. There was a significant dose, time, and dose-time interaction such that evaporative loss was dependent on initial PFC volume and time after fill (P < 0.05). Evaporative loss rate decreased earlier at lower doses. The percentage of initial volume lost to evaporation over time was inversely related to dose and could not be predicted by decreasing % PFC saturations, independent of dose. Evaporative loss should be considered to optimize both the application of PFC to the lung and replacement dosing during partial liquid ventilation. PMID- 11015135 TI - Modern statistical techniques for the analysis of longitudinal data in biomedical research. AB - Longitudinal study designs in biomedical research are motivated by the need or desire of a researcher to assess the change over time of an outcome and what risk factors may be associated with the outcome. The outcome is measured repeatedly over time for every individual in the study, and risk factors may be measured repeatedly over time or they may be static. For example, many clinical studies involving chronic obstructive pulmonary disease (COPD) use pulmonary function as a primary outcome and measure it repeatedly over time for each individual. There are many issues, both practical and theoretical, which make the analysis of longitudinal data complicated. Fortunately, advances in statistical theory and computer technology over the past two decades have made techniques for the analysis of longitudinal data more readily available for data analysts. The aim of this paper is to provide a discussion of the important features of longitudinal data and review two popular modern statistical techniques used in biomedical research for the analysis of longitudinal data: the general linear mixed model, and generalized estimating equations. Examples are provided, using the study of pulmonary function in cystic fibrosis research. PMID- 11015136 TI - Teen-age smoking. PMID- 11015137 TI - Wheezing as the sole clinical manifestation of cor triatriatum. AB - Cardiac malformations involving low-pressure chambers (i.e., either of the atria) are more often diagnosed later in life than lesions that involve high-pressure systems such as ventricular septal defects or persistent ducti arteriosi. Patients with congenital heart disease involving the atria may present only symptoms suggesting lung disease. We report on a child with recurrent episodes of wheezing, which did not respond to albuterol nebulizations and intravenous corticosteroids; he was subsequently found to have cor triatriatum. When a patient suffers from recurrent episodes of lower pulmonary infection and wheezing, despite appropriate management for asthma, less common (including cardiac) causes should be considered. PMID- 11015139 TI - Selected abstracts PMID- 11015138 TI - Status asthmaticus treated by high-frequency oscillatory ventilation. AB - We present a 2.5-year-old girl in severe asthma crisis who clinically deteriorated on conventional mechanical ventilation, but was successfully ventilated with high-frequency oscillatory ventilation (HFOV). Although HFOV is accepted as a technique for managing pediatric respiratory failure, its use in obstructive airway disease is generally thought to be contraindicated because of the risk of dynamic air-trapping. However, we suggest that obstructive airway disease can safely be managed with HFOV, provided certain conditions are met. These include the application of sufficiently high mean airway pressures to open and stent the airways ("an open airway strategy"), lower frequencies to overcome the greater attenuation of the oscillatory waves in the narrowed airways, permissive hypercapnia to enable reducing pressure swings as much as possible, longer expiratory times, and muscle paralysis to avoid spontaneous breathing. PMID- 11015140 TI - Elementary school absenteeism and air pollution. AB - This study assessed the association between ambient air pollution and daily elementary school absenteeism in Washoe County, NV, between 1996 and 1998. All 57 elementary schools in Washoe County in northern Nevada were included in the data set. There was a total of 27,793 student enrollments during this study period. The daily average absence rate was 5.09% (+/-1.54%). Air pollutant values including PM(10), O(3), and CO were obtained from seven air monitoring stations. Weather variables were collected from five of seven stations and from the Western Regional Climate Center. The daily average concentrations of PM(10), CO, and O(3) were 32.44 microg/m(3), 2.73 ppm, and 37.45 ppb, respectively. Student absenteeism was regressed on the three air pollutants, weather variables, and other confounding factors, using autoregression analysis. After adjusting for the effects of weather variables, day of the week, month, and holiday indicators, and time trend, we found that CO and O(3) were statistically significant predictors of daily absenteeism in elementary schools. For every 1.0 ppm and 50 ppb increase in CO and O(3), the absence rate would increase 3.79% (95% CI 1.04-6.55%) and 13.01% (95% CI 3.41-22.61%), respectively. However, PM(10) values were negatively correlated with school absenteeism. PMID- 11015141 TI - Subchronic silica exposure enhances respiratory defense mechanisms and the pulmonary clearance of Listeria monocytogenes in rats. AB - Both Listeria monocytogenes infection and silica exposure have been shown to significantly alter immune responses. In this study, we evaluated the effect of preexposure to silica on lung defense mechanisms using a rat pulmonary L. monocytogenes infection model. Male Sprague-Dawley rats were instilled intratracheally with saline (vehicle control) or silica using either an acute treatment regimen (5 mg/kg; 3 days) or a subchronic treatment protocol (80 mg/kg; 35 days). At 3 or 35 days after silica instillation, the rats were inoculated intratracheally with either approximately 5000 or 500,000 L. monocytogenes. At 3, 5, and 7 days postinfection, the left lung was removed, homogenized, and cultured on brain heart infusion agar at 37 degrees C. The numbers of viable L. monocytogenes were counted after an overnight incubation. Bronchoalveolar lavage (BAL) was performed on the right lungs, and BAL cell differentials, acellular lactate dehydrogenase (LDH) activity and albumin content were determined. Alveolar macrophage (AM) chemiluminescence (CL) and phagocytosis were assessed as a measure of macrophage function. Lung-associated lymph nodes were removed, and lymphocytes were recovered and differentiated. Preexposure to silica significantly increased the pulmonary clearance of L. monocytogenes as compared to saline controls. Exposure to silica caused significant increases in BAL neutrophils, LDH and albumin, and lymph-nodal T cells and natural killer (NK) cells in infected and noninfected rats. CL and phagocytosis were also elevated in silica-treated rats. In summary, the results demonstrated that exposure of rats to silica enhanced pulmonary immune responses, as evidenced by increases in neutrophils, NK cells, T lymphocytes, and macrophage activation. These elevations in pulmonary immune response are likely responsible for the increase in pulmonary clearance of L. monocytogenes observed with preexposure to silica. PMID- 11015142 TI - Estimation of dissolution rate from in vivo studies of synthetic vitreous fibers. AB - Although the dissolution rate of a fiber was originally defined by a measurement of dissolution in simulated lung fluid in vitro, it is feasible to determine it from animal studies as well. The dissolution rate constant for a fiber may be extracted from the decrease in long fiber diameter observed in certain intratracheal instillation experiments or from the observed long fiber retention in short-term biopersistence studies. These in vivo dissolution rates agree well with those measured in vitro for the same fibers. For those special types of fibers, the high-alumina rock wool fibers that could not be measured in vitro, the method provides a way of obtaining a chemical dissolution rate constant from an animal study. The inverse of the in vivo dissolution rate, the fiber dissolution time, correlates well with the weighted half life of long fibers in a biopersistence study, and the in vivo dissolution rate may be estimated accurately from this weighted half-life. PMID- 11015143 TI - Quantitative analysis of potential transfer of continuous glass filament from eclipse prototype 9-014 cigarettes. AB - This study was designed to determine if the Eclipse prototype 9-014 cigarettes, which use a special form of continuous glass filament (CGF) as an insulator around the carbon heat source, yield CGFs via mainstream smoke. A previously developed method (Higuchi et al., 2000) that employed electrostatic precipitation with a greater than 99% collection efficiency of mass-was used to capture CGFs transferred to mainstream smoke. The cigarettes were smoked using an exaggerated puffing condition more than twice the Federal Trade Commission (FTC) standard. Prior to smoking, cigarettes were subjected to handling procedures that simulated commercial shipping conditions. Using a modified standard addition method, and utilizing a mixture of water and glycerol as a mock condensate, CGFs were intentionally added to a series of (mock condensate) samples to develop knowledge of CGF recovery efficiency. The linear regression model of the recovered CGFs demonstrated a recovery efficiency of 86%. This efficiency rate was applied to the number of CGFs recovered from samples of smoke condensate and associated background samples. The number of CGFs in smoke condensate collected from the Eclipse 9-014 prototype was approximately 0.32 +/- 0.17 CGFs per cigarette (+/- standard deviation), including the background counts of CGFs, and 0.16 CGFs per cigarette when corrected for background contributions. The number of CGFs found in the smoke condensates for this prototype was statistically (p =.00031) distinguishable from zero and background in these experiments. The low number of CGFs seen in the transfer data from this prototype studied, the unique physical characteristics of the filaments (e.g., controlled physical dimensions), and the absence of biological activity of similar glass filaments/fibers indicate that biologically significant exposure to the Eclipse smoker does not occur. PMID- 11015144 TI - Safety assessment of continuous glass filaments used in eclipse. AB - Eclipse is a cigarette that produces smoke by primarily heating, rather than burning, tobacco. The Eclipse heat source assembly employs a continuous filament glass mat jacket to insulate the heat source. The glass mat insulator is composed of continuous glass filaments and a binder. The purpose of this article is to address the potential toxicological significance of the continuous glass filaments under the conditions of intended use. Transfer data and the unique physical characteristics of the filaments demonstrate that significant exposure of the smoker will not occur. The available environmental survey data clearly demonstrate that Eclipse smokers are extremely unlikely to be exposed to continuous glass filaments at a level that represents a biologically significant increase over background exposure to glass fibers. The chemical composition of the continuous glass filaments used in Eclipse is generally similar to C-glass fiber compositions such as MMVF 11 that have failed to produce either tumors or fibrosis in chronic inhalation studies conducted in rats. In vitro dissolution data demonstrate that the continuous glass filaments used in Eclipse are more soluble than biologically active fibers such as rock wool (MMVF 21) or asbestos. However, the continuous glass filaments used in Eclipse were not as soluble in simulated extracellular lung fluid as representative C-glass fibers (MMVF 10 and MMVF 11). In brief, exposure of Eclipse smokers to continuous glass filaments is extremely unlikely to occur at a level that may be construed to be of biological significance. PMID- 11015145 TI - Modified wick method using Weck-Cel sponges for collection of human rectal secretions and analysis of mucosal HIV antibody. AB - Weck-Cel sponges were examined for suitability as an absorbent material for nontraumatic collection of rectal secretions in humans. Sponges were tested in vitro and determined by quantitative enzyme-linked immunosorbent assay (ELISA) to be capable of releasing 100% of absorbed albumin and all immunoglobulin subtypes after treatment with detergent-supplemented buffer. Protein composition in rectal secretions collected from normal women with dry sponges (DS) or with sponges previously softened by moistening with saline (MS) was subsequently compared. DS secretions showed evidence of contamination with blood and interstitial fluid derived albumin, immunoglobulin G (IgG), and monomeric IgA. MS secretions appeared to represent local mucosal secretions more accurately because they contained negligible blood, a greater percentage of secretory IgA within the total IgA, and both lower albumin/IgG ratios and more dramatic alterations in IgG subclass distribution compared with corresponding serum. Anti-HIV IgG, IgM, IgA, and antibodies with secretory component could be demonstrated by ELISA in rectal secretions collected with moist sponges from 8 of 8, 1 of 8, 5 of 8, and 3 of 8 HIV-infected women, respectively. The data show that Weck-Cel sponges, if premoistened, can be used to collect rectal fluids nontraumatically and to obtain quantitative information about concentrations of immunoglobulins and specific antibodies on rectal mucosal surfaces. PMID- 11015146 TI - Absence of association between individual thymidine analogues or nonnucleoside analogues and lipid abnormalities in HIV-1-infected persons on initial therapy. AB - Changes in levels of triglycerides and cholesterol during antiretroviral therapy raise concerns regarding an increased future risk of atherogenic disease and may precede the appearance of fat redistribution. Hypotheses regarding the impact of nucleoside analogues on adipocytes provide a possible explanation for metabolic and clinical fat disturbances. It is unclear whether the choice of nucleoside analogue combination or coadministration of nonnucleoside agents influences change in lipids. We performed a cross-sectional analysis of 135 persons receiving their first nucleoside analogue plus nonnucleoside-based combination antiretroviral regimen for at least 1 month and for whom cholesterol and triglyceride values were available on therapy. Univariate and multivariate regression models were used to explore the relation between cholesterol and triglycerides, as continuous variables with other variables. Both significant and nonsignificant variables from univariate analyses were evaluated in multivariate models to limit possible confounders. No association with drug choice was observed, either when comparing thymidine analogues (stavudine or zidovudine), all nucleoside analogue combinations or choice of either efavirenz or nevirapine as nonnucleoside. Age and triglyceride levels were found in a multivariate analysis to be associated with higher cholesterol. Only higher cholesterol was associated with higher triglyceride levels. In conclusion, no differences were observed between choice of drug or combination on cholesterol or triglyceride values during therapy. Older individuals may be more likely to have elevated cholesterol values. PMID- 11015147 TI - Differences between women and men in adverse events and CD4+ responses to nucleoside analogue therapy for HIV infection. The Aids Clinical Trials Group 175 Team. AB - OBJECTIVE: To prospectively examine differences in baseline characteristics and study outcomes between HIV-infected women and men during a clinical trial of nucleoside analogue therapy. METHODS: ACTG 175 randomized HIV-infected patients with CD4+ counts between 200 and 500 cells/mm3 to one of four nucleoside analogue regimens: zidovudine (ZDV), didanosine (ddI), ZDV + ddI, or ZDV + zalcitabine (ddC). Differences in time to first dose modification, voluntary withdrawal, development of toxicity and symptomatology, and AIDS progression were compared by gender. RESULTS: The study included 438 women and 2029 men. Baseline values of HIV RNA plasma concentrations were significantly lower for women (0.3 log10) than men in a subset of patients in whom assays were taken and this difference persisted after adjustment for CD4+ count. Women reported reducing dosage and discontinue ddI-containing regimens more frequently than men did; adjustment for weight did not completely explain this difference. Women were at lower risk than men for progression to a study endpoint (19% of women versus 24% of men; p <.0001). Among those antiretroviral-naive study subjects receiving ZDV, men were four times more likely to progress to a study endpoint than women. CONCLUSIONS: Differences in pretreatment characteristics and on study experiences were demonstrated between women and men enrolled in this clinical trial. The suggestion of a gender difference in response to ZDV monotherapy by antiretroviral-naive study subjects and the lower baseline values for HIV RNA in women compared with those in men provides evidence for gender differences in the relationship between virus replication, CD4+ decline, and responses to nucleoside analogue therapy. PMID- 11015148 TI - Effects of lipolysis or heat treatment on HIV-1 provirus in breast milk. AB - BACKGROUND: Transmission of HIV-1 infection through breastfeeding is associated with integrated DNA (provirus) in milk cells. Reduction of HIV-1 DNA in milk may lessen infectivity. PURPOSE: To investigate efficacy of two methods available in developing countries to reduce HIV-1 proviral DNA in breast milk. METHODS: Methods simulated field conditions; milk was heated by bringing it to a boil, for instance, over a cooking fire, and lipolysis was done at room temperature. Four HIV-positive pregnant women were recruited for this pilot study, instructed to feed formula exclusively, and to stimulate milk production using pumping. Milk was collected twice weekly for 3 weeks and analyzed qualitatively for HIV-1 proviral DNA by polymerase chain reaction at three stages: 1) fresh, 2) after standing for 6 hours, and 3) after having been brought to the boiling point. RESULTS: Seventeen samples from 4 mothers were analyzed. Fifteen of 17 fresh samples (88%) had measurable HIV-1 proviral DNA despite all mothers' having had low or undetectable plasma viral loads. Lipolysis (standing at room temperature) for 6 hours did not destroy proviral DNA: 6 of 7 samples (86%) tested positive for DNA after lipolysis. No samples of milk (n = 8) brought to a boil were positive for HIV-1 proviral DNA (p <.0001). CONCLUSIONS: This preliminary evidence suggests that inherent lipolytic activity of fresh breast milk is inadequate for destruction of HIV-1; bringing breast milk to a boil may result in decreased HIV-1 infectivity; and breast milk cell-associated HIV-1 may not reflect plasma viral load. Nutritional value or possible bacterial contamination of milk treated in this manner was not assessed. PMID- 11015149 TI - Cell-free virus in breast milk of HIV-1-seropositive women. AB - To examine the prevalence, quantification, and factors that influence HIV in the cell-free compartment of breast milk, we performed reverse transcription polymerase chain reaction (RT-PCR) on samples obtained from HIV-1-infected study subjects. Virus was detected in 86 of 136 samples (63.2%) from 79 study subjects. HIV RNA quantity ranged from undetectable to 227,586 copies/ml. Prevalence and mean viral load were not affected by postnatal ages or maternal vitamin A supplementation. Among study subjects with multiple samples, breast milk viral load did not change at different postnatal ages. Breast milk viral load correlated positively with plasma viral load (r = 0.47; p =.005) and negatively with maternal CD4 count at entry to the study (r = -0.26; p =.02). Mothers of HIV infected children had a higher proportion of detectable HIV RNA in their breast milk than mothers of uninfected children (p =.03) and higher mean log10 HIV RNA quantities (p =.04). In a multivariate logistic regression model, log10 HIV RNA quantity in breast milk was significantly associated with the risk of mother child transmission (odds ratio [OR], 2.82; 95% confidence interval [CI], 1.22 6.51). Thus, prevention and treatment of opportunistic infections and of mastitis and early weaning may be important elements of a public health policy that is relevant to women in developing countries with HIV infection. Where available, antiretrovirals may also have an impact on opportunistic infections and mastitis. PMID- 11015150 TI - Reduced toxicity with gradual initiation of trimethoprim-sulfamethoxazole as primary prophylaxis for Pneumocystis carinii pneumonia: AIDS Clinical Trials Group 268. AB - Trimethoprim-sulfamethoxazole (TMP/SMX) is recognized as the superior agent for Pneumocystis carinii pneumonia (PCP) prophylaxis but a high incidence of adverse drug reactions, which may be due to toxic drug metabolites, limits its use. AIDS Clinical Trials Group protocol 268 was a randomized, double-blind, controlled two arm trial designed to determine whether gradual initiation of TMP/SMX suspension reduced the incidence of treatment-limiting adverse drug reactions compared with routine initiation of double-strength (DS; 160 mg/800 mg) tablets. In all, 372 HIV-1-infected study subjects with a CD4+ cell count <250 x 10 cells/mm3 who had not previously received TMP/SMX for PCP prophylaxis were randomized to receive either daily TMP/SMX DS tablets or a gradually increasing dose of TMP/SMX suspension. The suspension dose was increased to reach the equivalent of a DS tablet by study day 13. During the first 2 weeks, study subjects also received a matching placebo tablet/suspension. After week 2, all study subjects received TMP/SMX tablets for the next 10 weeks. There were significantly fewer study subjects who discontinued prophylaxis during the first 12 weeks when TMP/SMX therapy was initiated gradually (17%) than when initiated in DS tablet formulation (33%) (p =.0002). Gradual initiation was also associated with significantly fewer adverse drug reactions. Gradual initiation of TMP/SMX for primary PCP prophylaxis reduces the incidence of its treatment-limiting adverse effects. PMID- 11015151 TI - Oral ganciclovir systemic exposure is enhanced in HIV-infected patients with diarrhea and weight loss. AB - OBJECTIVE: To determine whether diarrhea and intestinal malabsorption during HIV infection alter oral ganciclovir systemic exposure. METHODS: We studied the oral disposition of ganciclovir in 42 HIV-infected patients stratified into three groups: A (n = 15), HIV (stage A and B); B (n = 13), AIDS (stage C); and C (n = 14), AIDS with chronic diarrhea and wasting syndrome (10% or more weight loss). Each patient was evaluated for nutritional (body mass index, serum albumin and transferrin), immunologic (CD4 count, plasma viral load) and intestinal status (D xylose test, fecal fat and nitrogen excretion, and intestinal permeability). Following an overnight fast, 1 g oral ganciclovir was given to patients. Six blood samples were collected over 24 hours. Serum was analyzed for ganciclovir by high performance liquid chromatography. Drug disposition was characterized using a population pharmacokinetic approach. RESULTS: Mean intestinal permeability increased as HIV disease progressed (0. 05, 0.1, and 0.2 for groups A, B, and C, respectively). Average weight-adjusted maximum concentration (Cmax) in group C was twofold more than that in group A and B patients (12.5 versus 6 and 6.4 ng/ml/kg), and average area under the curve (AUC0-infinity) was threefold greater in group C patients (193 versus 59 and 65 ng. hour/ml/kg in groups A and B, respectively). Mean oral clearance was threefold lower in group C (96 versus 258 and 212 L/hour in groups A and B, respectively). CONCLUSION: Because systemic exposure of oral ganciclovir is enhanced in AIDS patients with diarrhea and wasting syndrome, oral ganciclovir therapy may benefit these patients. PMID- 11015152 TI - Model of HIV-1 disease progression based on virus-induced lymph node homing and homing-induced apoptosis of CD4+ lymphocytes. AB - Several proposed theories have described the progression of HIV infection. Even so, no concrete evidence supports any as comprehensive, including, for example, why the CD4+ T-cell counts fall from 1000/mm3 of blood to roughly 100/mm3 over an average 10-year period, whereas concomitant viral loads are relatively constant, increasing by several orders of magnitude in late-stage disease. Here, we develop and validate a theoretical model that altered lymphocyte circulation patterns between the lymph system and blood due to HIV-induced enhanced lymph-node homing and subsequent apoptosis of resting CD4+ T cells can explain many aspects of HIV 1 disease progression. These results lead to a recalculation of the CD4+ lymphocyte dynamics during highly active antiretroviral therapy, and also suggest new targets for therapy. PMID- 11015153 TI - Obtaining HIV test results with a home collection test kit in a community telephone sample. AB - OBJECTIVES: To test the feasibility of obtaining HIV test results by home collection kit from a probability telephone sample of men who have sex with men (MSM). METHODS: A quota sample of 615 MSM previously interviewed by the Urban Men's Health Study phone survey in Chicago, Los Angeles, New York City, and San Francisco were re-contacted and offered an HIV test using an oral specimen (Orasure) home collection kit. RESULTS: Eighty percent consented to be mailed a kit, and 84% returned a specimen, for a 67% participation rate. All self-reported HIV-positive persons tested positive (77 of 77); 4 of 266 (1.5%) with a prior negative test and 2 of 69 (2.9%) with no prior positive HIV test result. Participation was associated with self-reported prior HIV test status-HIV positive (83%), HIV-negative (68%), or no prior HIV test result (54%)-and marginally associated with New York City residence after adjustment for HIV status (odds ratio = 0.7; 95% confidence interval, 0.4-1.1; p =.08). CONCLUSIONS: These results suggest that urban MSM identified and interviewed by telephone will participate in home collection HIV testing. This methodology could be used to produce population-based estimates of HIV seroprevalence and seroincidence in MSM and could probably be extended to other populations and other viral infections. PMID- 11015155 TI - Clinical spectrum, morbidity, and mortality of acquired immunodeficiency syndrome in Taiwan: a 5-year prospective study. AB - The clinical spectrum of AIDS and changes of morbidity and mortality associated with HIV infection following initiation of highly active antiretroviral therapy (HAART) are rarely described in the less developed countries in the Asia-Pacific region. We prospectively observed on a follow-up basis 309 HIV-infected patients (82.8% with AIDS) at National Taiwan University Hospital in Taiwan, where highly active antiretroviral therapy (HAART) has been provided to all patients at no charge at any stage of HIV infection since April 1, 1997, to describe the spectrum of HIV-associated opportunistic diseases and evaluate changes of morbidity and mortality from June 24, 1994 through June 23, 1999. Of the patients, 59.3% at study entry had a CD4+ lymphocyte count of <50 cells/microliter. The five leading HIV-associated opportunistic infections included oroesophageal candidiasis (195 patients), Pneumocystis carinii pneumonia (93), tuberculosis (77), mucocutaneous herpes simplex infection (74), and cytomegalovirus diseases (73). The incidence rates of seven major AIDS-defining opportunistic diseases were declining though the changes of the relative proportions varied. The median duration of hospitalization decreased from 36 days in 1995 to 12 days in 1999 (p =.0001). Overestimated mortality rate declined from 148.4 per 100 patient-years in 1995 to 7.4 per 100 patient-years in 1999 (p =.0001) whereas the underestimated mortality rate declined from 110.5 to 5.39 per 100 patient-years (p =.0001). Risk ratio (RR) for mortality in patients who received HAART compared with those who did not was 0.410 (95% confidence interval [CI], 0.249-0.674; p =.0004) and the RR was 0.250 (95% CI, 0.127-0.492; p =.0001) when the analysis was limited to patients with an initial CD4+ lymphocyte count <100 cells/microliter and follow-up duration >30 days after adjusting for their age, gender, type of risk behavior, and CD4+ lymphocyte count. Morbidity and mortality were declining with each study year even in a population consisting mainly of patients at the advanced stage of HIV infection in Taiwan. Earlier diagnosis, accumulation of clinical experience, and use of HAART were associated with lower mortality rates. PMID- 11015154 TI - Cost-effective use of nevirapine to prevent vertical HIV transmission in sub Saharan Africa. AB - OBJECTIVE: To assess the cost-effectiveness of alternative strategies of nevirapine (NVP) administration to prevent vertical HIV transmission in sub Saharan Africa. DESIGN: A decision-analysis model was constructed to estimate the costs and effects of NVP-based prevention strategies for two separate groups of women: those who qualify for standard therapy by attending a 36-week prenatal visit, and those who do not qualify, owing to preterm delivery or lack of prenatal care. RESULTS: For women in prenatal care, mass provision of NVP without maternal serodiagnosis was found to yield greater health gains at an acceptable cost, compared with providing targeted therapy to only those women identified as seropositive. However, this conclusion was strongly contingent on several uncertain assumptions, most importantly the probability that a woman who does not know her serostatus will nonetheless adhere to therapy. Among those women who present for delivery without prior enrollment in a prenatal strategy, either late provision of maternal-infant NVP or treatment of only the infant would likely be a cost-effective alternative to the current practice of offering no preventive therapy. CONCLUSIONS: NVP intervention offers a cost-effective avenue for preventing vertical HIV transmission in sub-Saharan Africa. The optimal choice between mass therapy and targeted therapy cannot be confidently identified without information regarding adherence among women who do not know their serostatus. For women who do not receive NVP prenatally, treatment on presentation for delivery would be cost-effective even in the face of modest clinical efficacy. Clinical assessment of adherence to therapy among women who do not know their status and the field effectiveness of alternative approaches to NVP administration is urgently needed to allow identification of optimal prevention strategies. PMID- 11015156 TI - High-risk behaviors during incarceration in African-American men treated for HIV at three Los Angeles public medical centers. AB - OBJECTIVES: This paper describes research that examined the association between high-risk sexual and drug-using behaviors during incarceration and HIV infection for African-American men receiving HIV care at three public medical centers in Los Angeles County (LAC), California. METHODS: A case-control study was conducted in which 305 HIV-infected African-American men and 305 neighborhood controls, ages 20 to 49, were frequency-matched by age. RESULTS: After controlling for anal sex while not incarcerated, we found no association between anal sex during incarceration and HIV (odds ratio [OR], 1.1; 95% confidence interval [CI], 0.6 2.2). Among men with a history of incarceration (n = 332), the percentage reporting anal sex with men outside of incarceration (45%) was greater than those reporting anal sex while incarcerated (16%). Injection drug use (IDU) during incarceration was also not associated with HIV when controlling for IDU outside of incarceration (OR, 1.6; 95% CI, 0.5- 4.9). Increased time in jail or prison was associated with less HIV infection (p =.001). CONCLUSIONS: Although high-risk behaviors are more common in the community than in the incarcerated setting for this study group, incarcerated populations represent a high-risk group for whom access to prevention messages is limited. Periods of incarceration represent a unique opportunity to convey prevention messages that focus on high-risk behaviors outside the incarcerated setting. PMID- 11015157 TI - Anal sex among HIV-seronegative women at high risk of HIV exposure. The HIVNET Vaccine Preparedness Study 2 Protocol Team. AB - To assess the prevalence and the sociodemographic and behavioral correlates of anal sex in a cohort of HIV-seronegative U.S. women at high risk of HIV exposure, we administered a risk assessment using audio computer-assisted self-interview (A CASI). Of 1268 sexually active women, 432 (32%) reported anal sex in the previous 6 months. Compared with women who did not report anal sex, those who did had more unprotected vaginal sex (median of 11 versus 7 episodes; p <. 001) and a higher proportion of unprotected sexual (vaginal plus anal) episodes (median of 0.90 versus 0.81; p =.01). Anal sex was reported by higher proportions of women who did not always use condoms, who used crack in the past year, who were 0.2). These findings suggest that the association between meal ingestion and head-up tilt-table testing were additive and not synergistic. However, the proportion of participants with symptomatic hypotension increased during head-up tilt-table testing after meal ingestion (12% during preprandial testing and 22% during postprandial testing). Symptomatic hypotension tended to occur more often and sooner after meal ingestion than before meal ingestion (P = 0.03). CONCLUSIONS: Meal ingestion and head-up tilt-table testing are associated with increasing occurrences of symptomatic hypotension. After meal ingestion and head-up tilt table testing, 22% of functionally independent elderly persons had symptomatic hypotension. PMID- 11015167 TI - Diagnostic implications of elevated levels of smooth-muscle myosin heavy-chain protein in acute aortic dissection. The smooth muscle myosin heavy chain study. AB - BACKGROUND: A rapid 30-minute assay of circulating smooth-muscle myosin heavy chain protein has been developed as a biochemical diagnostic tool for aortic dissection. OBJECTIVE: To determine the sensitivity and specificity of this assay. DESIGN: Cross-sectional study. SETTING: 8 major cardiovascular centers in Japan. PATIENTS: 95 patients with acute aortic dissection, 48 patients with acute myocardial infarction, and 131 healthy volunteers. MEASUREMENTS: Levels of circulating smooth-muscle myosin heavy-chain protein. RESULTS: Patients with acute aortic dissection who presented within 3 hours after onset had elevated levels of circulating smooth-muscle myosin heavy-chain protein. In these patients, the assay had a sensitivity of 90.9%, a specificity of 98% compared with healthy volunteers, and a specificity of 83% compared with patients who had acute myocardial infarction; the clinical decision limit was 2.5 microgram/L. All patients with proximal lesions had elevated levels of smooth-muscle myosin heavy chain protein, and only patients with distal lesions had decreased levels (<2.5 microgram/L). CONCLUSIONS: Levels of smooth-muscle myosin heavy-chain protein can be used to diagnose aortic dissection soon after symptom onset. The assay had the greatest diagnostic value in patients with proximal lesions. PMID- 11015168 TI - Update in gastroenterology. PMID- 11015169 TI - "The lower the better" in hypercholesterolemia therapy: a reliable clinical guideline?. AB - Since the publication of the second set of guidelines by the National Cholesterol Education Program, a solid body of data from landmark clinical studies has demonstrated that reduction in low-density lipoprotein (LDL) cholesterol with 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitor ("statin") therapy sharply diminishes the risk for coronary artery disease. These trials include the Scandinavian Simvastatin Survival Study, the West of Scotland Coronary Prevention Study, the Air Force/Texas Coronary Atherosclerosis Prevention Study, the Cholesterol and Recurrent Events investigation, and the Long-Term Intervention with Pravastatin in Ischaemic Disease trial. Coronary event rates and, in some cases, all-cause mortality decreased significantly after about 5 years of statin therapy in patients at risk for and those who had coronary artery disease at baseline. In contrast, recent subgroup analyses of these pivotal studies have in the aggregate challenged the premise that lower LDL cholesterol levels necessarily lead to further declines in risk for coronary artery disease, particularly among the patients most likely to be seen by the clinician: those with moderately elevated or normal cholesterol profiles. Indeed, when LDL cholesterol levels are in this range, further lowering with statin therapy elicits diminishing returns in terms of coronary event rates. These findings are readily accommodated by the curvilinear, or log-linear, model between serum cholesterol level and risk for coronary artery disease, which is predicated on data from large epidemiologic studies. In light of the current climate involving competing health care costs, the pursuit of progressively diminishing returns in terms of reductions in coronary artery disease risk through more aggressive lowering of LDL cholesterol levels appears to be unwarranted. Until data are published from ongoing randomized, clinical trials that can more effectively resolve the clinical utility of aggressive lipid-lowering strategies to improve coronary event rates, a prudent, evidence-based strategy seems warranted. PMID- 11015170 TI - Autologous stem-cell transplantation in ovarian cancer: is more better? PMID- 11015171 TI - The Face of Evil. PMID- 11015172 TI - On being a doctor. Alternative healer. PMID- 11015173 TI - Responding to intractable terminal suffering. PMID- 11015174 TI - Responding to intractable terminal suffering. PMID- 11015175 TI - Responding to intractable terminal suffering. PMID- 11015176 TI - Responding to Intractable Terminal Suffering. PMID- 11015177 TI - Palliative treatment of last resort and assisted suicide. PMID- 11015178 TI - Palliative Treatment of Last Resort and Assisted Suicide. PMID- 11015180 TI - Palliative Treatment of Last Resort and Assisted Suicide. PMID- 11015179 TI - Palliative treatment of last resort and assisted suicide. PMID- 11015181 TI - Publication of papers on assisted suicide and terminal sedation. PMID- 11015182 TI - Publication of Papers on Assisted Suicide and Terminal Sedation. PMID- 11015184 TI - Medicine and the Movies: Lorenzo's Oil at Century's End. PMID- 11015183 TI - Publication of Papers on Assisted Suicide and Terminal Sedation. PMID- 11015185 TI - Parting Company: Understanding the Loss of a Loved One. The Caregiver's Journey. PMID- 11015186 TI - Transfusion Therapy: Clinical Principles and Practice. PMID- 11015187 TI - Mechanism of transmembrane signaling: insulin binding and the insulin receptor. AB - Transmembrane signaling via receptor tyrosine kinases generally requires oligomerization of receptor monomers, with the formation of ligand-induced dimers or higher multimers of the extracellular domains of the receptors. Such formations are expected to juxtapose the intracellular kinase domains at the correct distances and orientations for transphosphorylation. For receptors of the insulin receptor family that are constitutively dimeric, or those that form noncovalent dimers without ligands, the mechanism must be more complex. For these, the conformation must be changed by the ligand from one that prevents activation to one that is permissive for kinase phosphorylation. How the insulin ligand accomplishes this action has remained a puzzle since the discovery of the insulin receptor over 2 decades ago, primarily because membrane proteins in general have been refractory to structure determination by crystallography. However, high-resolution structural evidence on individual separate subdomains of the insulin receptor and of analogous proteins has been obtained. The recently solved quaternary structure of the complete dimeric insulin receptor in the presence of insulin has now served as the structural envelope into which such individual domains were fitted. The combined structure has provided answers on the details of insulin/receptor interactions in the binding site and on the mechanism of transmembrane signaling of this covalent dimer. The structure explains many observations on the behavior of the receptor, from greater or lesser binding of insulin and its variants, point and deletion mutants of the receptor, to antibody-binding patterns, and to the effects on basal and insulin stimulated autophosphorylation under mild reducing conditions. PMID- 11015188 TI - Metal-phosphate interactions in the hammerhead ribozyme observed by 31P NMR and phosphorothioate substitutions. AB - The hammerhead ribozyme is a catalytic RNA that requires divalent metal cations for activity under moderate ionic strength. Two important sites that are proposed to bind metal ions in the hammerhead ribozyme are the A9/G10.1 site, located at the junction between stem II and the conserved core, and the scissile phosphate (P1.1). (31)P NMR spectroscopy in conjunction with phosphorothioate substitutions is used in this study to investigate these putative metal sites. The (31)P NMR feature of a phosphorothioate appears in a unique spectral window and can be monitored for changes upon addition of metals. Addition of 1-2 equiv of Cd(2+) to the hammerhead with an A9-S(Rp) or A9-S(S)(Rp) substitution results in a 2-3 ppm upfield shift of the (31)P NMR resonance. In contrast, the P1.1-S(Rp) and P1.1 S(Sp) (31)P NMR features shift slightly and in opposite directions, with a total change in delta of 90-fold more rapidly and to a higher yield than an adjacent adenosine in the same RNA structure. This reactivity is minimally dependent on the base pairing partner of the edited nucleotide; adenosine at the editing site in the naturally occurring A.C mismatch is deaminated to approximately the same extent and only 4 times faster than adenosine in an A.U base pair at this site. A steady-state rate analysis at a saturating concentration of the most rapidly processed substrate indicates that product formation is linear with time through at least three turnovers with a slope of 13 +/- 1.5 nM.min(-1) at 30 nM ADAR2 for a k(ss) = 0.43 +/- 0.05 min( 1). In addition, ADAR2 induces a 3.3-fold enhancement in fluorescence intensity and a 14 nm blue shift in the emission maximum of a duplex substrate with 2 aminopurine located at the editing site, consistent with a mechanism whereby ADAR2 flips the reactive nucleotide out of the double helix prior to deamination. PMID- 11015204 TI - Differential incision of bulky carcinogen-DNA adducts by the UvrABC nuclease: comparison of incision rates and the interactions of Uvr subunits with lesions of different structures. AB - The UvrABC nuclease system from Escherichia coli removes DNA damages induced by a wide range of chemical carcinogens with variable efficiencies. The interactions with UvrABC proteins of the following three lesions site-specifically positioned in DNA, and of known conformations, were investigated: (i) adducts derived from the binding of the (-)-(7S,8R,9R,10S) enantiomer of 7,8-dihydroxy-9, 10-epoxy 7,8,9,10-tetrahydrobenzo[a]pyrene [(-)-anti-BPDE] by cis-covalent addition to N(2)-2'-deoxyguanosine [(-)-cis-anti-BP-N(2)-dG], (ii) an adduct derived from the binding of the (+)-(1R,2S,3S,4R) enantiomer of 1,2-dihydroxy-3,4-epoxy-1,2,3, 4 tetrahydro-5-methylchrysene [(+)-anti-5-MeCDE] by trans addition to N(2)-2' deoxyguanosine [(+)-trans-anti-MC-N(2)-dG], and (iii) a C8-2'-deoxyguanosine adduct (C8-AP-dG) formed by reductively activated 1-nitropyrene (1-NP). The influence of these three different adducts on UvrA binding affinities, formation of UvrB-DNA complexes by quantitative gel mobility shift analyses, and the rates of UvrABC incision were investigated. The binding affinities of UvrA varied among the three adducts. UvrA bound to the DNA adduct (+)-trans-anti-MC-N(2)-dG with the highest affinity (K(d) = 17 +/- 2 nM) and to the DNA containing C8-AP-dG with the least affinity (K(d) = 28 +/- 1 nM). The extent of complex formation with UvrB was also the lowest with the C8-AP-dG adduct. 5' Incisions occurred at the eighth phosphate from the modified guanine. The major 3' incision site corresponded to the fifth phosphodiester bond for all three adducts. However, additional 3' incisions were observed at the fourth and sixth phosphates in the case of the C8-AP-dG adduct, whereas in the case of the (-)-cis-anti-BP-N(2)-dG and (+)-trans-anti-MC-N(2)-dG lesions additional 3' cleavage occurred at the sixth and seventh phosphodiester bonds. Both the initial rate and the extent of 5' and 3' incisions revealed that C8-AP-dG was repaired less efficiently in comparison to the (-)-cis-anti-BP-N(2)-dG and (+)-trans-anti-MC-N(2)-dG containing DNA adducts. Our study showed that UvrA recognizes conformational changes induced by structurally different lesions and that in certain cases the binding affinities of UvrA and UvrB can be correlated with the incision rates. The size of the bubble formed around the damaged site with mismatched bases also appears to influence the incision rates. A particularly noteworthy finding in this study is that UvrABC repair of a substrate with no base opposite C8-AP-dG was quite inefficient as compared to the same adduct with a C opposite it. These findings are discussed in terms of the available NMR solution structures. PMID- 11015205 TI - Evaluation of the effectiveness of DNA-binding drugs to inhibit transcription using the c-fos serum response element as a target. AB - Previous work has demonstrated that sequence-selective DNA-binding drugs can inhibit transcription factors from binding to their target sites on gene promoters. In this study, the potency and effectiveness of DNA-binding drugs to inhibit transcription were assessed using the c-fos promoter's serum response element (SRE) as a target. The drugs chosen for analysis included the minor groove binding agents chromomycin A(3) and Hoechst 33342, which bind to G/C-rich and A/T-rich regions, respectively, and the intercalating agent nogalamycin, which binds G/C-rich sequences in the major groove. The transcription factors targeted, Elk-1 and serum response factor (SRF), form a ternary complex (TC) on the SRE that is necessary and sufficient for induction of c-fos by serum. The drugs' abilities to prevent TC formation on the SRE in vitro were nogalamycin > Hoechst 33342 > chromomycin. Their potencies in inhibiting cell-free transcription and endogenous c-fos expression in NIH3T3 cells, however, were chromomycin > nogalamycin > Hoechst 33342. The latter order of potency was also obtained for the drugs' cytotoxicity and inhibition of general transcription as measured by [(3)H]uridine incorporation. These systematic analyses provide insight into how drug and transcription factor binding characteristics are related to drugs' effectiveness in inhibiting gene expression. PMID- 11015206 TI - Sequence determinants for the recognition of the fork junction DNA containing the -10 region of promoter DNA by E. coli RNA polymerase. AB - It has been recently suggested that E. coli RNA polymerase can specifically recognize a fork junction DNA structure, suggesting a possible role for such interaction in promoter DNA melting [Guo, Y., and Gralla, J. D. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 11655-11660]. We have determined here quantitatively, using a site-specific binding assay, the effects of base substitutions within the conserved -10 hexamer in the context of a short fork junction DNA on binding to RNA polymerase. Adenine at position -11 and thymine at position -7 were found to be critical for sequence-specific recognition of the DNA. The identities of bases at positions -9 and -8 were found to be not important for the binding whereas replacement of bases at positions -12 and -10 had a mild negative effect on the binding affinity. It was found that for the binding of fork DNA to RNA polymerase, specific sequence recognition was more important than specific recognition of fork junction DNA structure. The pattern of relative importance of bases in the -10 region for binding RNA polymerase was generally consistent with the sequence conservation pattern observed in nature where positions -11 and -7 are the most conserved. Binding experiments with a series of adenine analogues at position -11 revealed that the N1 nitrogen of adenine was a critical determinant for the preference of the adenine at this position, suggesting a mechanism for the nucleation of promoter DNA melting initiation in which RNA polymerase destabilizes duplex DNA by directly competing with the thymine of the A-T base pair for hydrogen bonding to the N1 position of the -11 nontemplate strand adenine. PMID- 11015207 TI - Intervesicle cross-linking with integrin alpha IIb beta 3 and cyclic-RGD lipopeptide. A model of cell-adhesion processes. AB - We report the synthesis of a new integrin alpha(IIb)beta(3)-specific cyclic hexapeptide that contains an Arg-Gly-Asp (RGD) sequence and is coupled to a dimyristoylthioglyceryl anchor. We demonstrate that this ligand is useful to study specific integrin binding to membrane surfaces. With the help of biotinylated analogues of the peptide, a spacer of optimal length between the peptide and lipid moieties was searched for by evaluating the binding strength with an enzyme-coupled immunosorbent assay (ELISA) and by surface plasmon resonance (SPR). It was found to be strongly dependent on the length of the spacer introduced between the biotin and peptide moieties of the ligands, which consisted either of epsilon-aminohexanoic acid (epsilonAhx) or of epsilonAhx with two additional glycine units. Best results were obtained with c[Arg-Gly-Asp-D-Phe Lys(Biot-Ahx-Gly-Gly)-Gly-] with dissociation constants of K(D) = 0.158 microM from ELISA and K(D) = 1.1 microM from SPR measurements. The analogous lipopeptide, c[Arg-Gly-Asp-D-Phe-Lys([dimyristoyl-3-thioglyceryl-succinimido propanoyl]Ahx-Gly-Gly)-Gly], was used as a membrane-anchored integrin ligand. It is shown by fluorescence microscopy and cryo electron microscopy that integrin reconstituted into phospholipid vesicles binds to vesicles decorated with the lipopeptide, forming regularly spaced bridges between the two kinds of vesicles. The novel integrin-specific ligand allows establishment of new model systems for systematic studies of the self-organization of integrin clusters and focal adhesion complexes. PMID- 11015208 TI - Stathmin slows down guanosine diphosphate dissociation from tubulin in a phosphorylation-controlled fashion. AB - Stathmin is an important protein that interacts with tubulin and regulates microtubule dynamics in a phosphorylation-controlled fashion. Here we show that the dissociation of guanosine 5'-diphosphate (GDP) from beta-tubulin is slowed 20 fold in the (tubulin)(2)-stathmin ternary complex (T(2)S). The kinetics of GDP or guanosine 5'-triphosphate (GTP) dissociation from tubulin have been monitored by the change in tryptophan fluorescence of tubulin upon exchanging 2-amino-6 mercapto-9-beta-ribofuranosylpurine 5'-diphosphate (S6-GDP) for tubulin-bound guanine nucleotide. At molar ratios of stathmin to tubulin lower than 0.5, biphasic kinetics were observed, indicating that the dynamics of the complex is extremely slow, consistent with its high stability. The method was used to characterize the effects of phosphorylation of stathmin on its interaction with tubulin. The serine-to-glutamate substitution of all four phosphorylatable serines of stathmin (4E-stathmin) weakens the stability of the T(2)S complex by about 2 orders of magnitude. The phosphorylation of serines 16 and 63 in stathmin has a more severe effect and weakens the stability of T(2)S 10(4)-fold. The rate of GDP dissociation is lowered only 7-fold and 4-fold in the complexes of tubulin with 4E-stathmin and diphosphostathmin, respectively. Sedimentation velocity studies support the conclusions of nucleotide exchange data and show that the T(2)S complexes formed between tubulin and 4E-stathmin or diphosphostathmin are less compact than the highly stable T(2)S complex. The correlation between the effect of phosphorylation of stathmin on the stability of T(2)S complex measured in vitro and on the function of stathmin in vivo is discussed. PMID- 11015209 TI - Secondary structure and oligomerization of the E. coli glycerol facilitator. AB - The Major Intrinsic Proteins are found throughout the bacterial, plant, and animal kingdoms and are responsible for the rapid transport of water and other small, polar solutes across membranes. The superfamily includes the aquaporins, the aquaglyceroporins, and the glycerol facilitators. We have overexpressed and purified the Escherichia coli inner membrane glycerol facilitator. Approximately 7.5 mg of 95% pure protein is obtained from 1 L of Escherichia coli cells using immobilized metal affinity chromatography. Well-resolved matrix-assisted laser desorption ionization mass spectra were obtained by solubilization of the protein in octyl-beta-D-glucopyranoside (M(r) = 33 650.3; error approximately 0.4%). The recombinant glycerol facilitator is inserted into the bacterial inner membrane, is functional, and is inhibited by HgCl(2). Polyacrylamide gel electrophoresis suggests that the facilitator is predominantly monomeric when solubilized with dodecyl-beta-D-maltoside, octyl-beta-D-glucopyranoside, and sodium dodecyl sulfate, but that it self-associates, forming soluble oligomers when urea is used during extraction. Similar oligomeric species are demonstrated to exist in the bacterial membrane by chemical cross-linking experiments. Circular dichroism analysis shows that the protein is predominantly alpha-helical. Helix content is significantly higher in protein prepared in the absence of urea (42-55%) than in its presence (32%). A possible role for the facilitator oligomers in interactions with, and regulation of, the glycerol kinase is discussed. PMID- 11015210 TI - Structural basis of the anionic interface preference and kcat* activation of pancreatic phospholipase A2. AB - Pancreatic phospholipase A(2) (PLA2) shows a strong preference for the binding to the anionic interface and a consequent allosteric activation. In this paper, we show that virtually all the preference is mediated through 3 (Lys-53, -56, and 120) of the 12 cationic residues of bovine pancreatic PLA2. The lysine-to methionine substitution enhances the binding of the enzyme to the zwitterionic interface, and for the K53,56,120M triple mutant at the zwitterionic interface is comparable to that for the wild type (WT) at the anionic interface. In the isomorphous crystal structure, the backbone folding of K53,56M K120,121A and WT are virtually identical, yet a significant change in the side chains of certain residues, away from the site of substitution, mostly at the putative contact site with the interface (i-face), is discernible. Such reciprocity, also supported by the spectroscopic results for the free and bound forms of the enzyme, is expected because a distal structural change that perturbs the interfacial binding could also affect the i-face. The results show that lysine-to-methionine substitution induces a structural change that promotes the binding of PLA2 to the interface as well as the substrate binding to the enzyme at the interface. The kinetic results are consistent with a model in which the interfacial Michaelis complex exists in two forms, and the complex that undergoes the chemical step is formed by the charge compensation of Lys-53 and -56. Analysis of the incremental changes in the kinetic parameters shows that the charge compensation of Lys-53 and -56 contributes to the activation and that of Lys-120 contributes only to the structural change that promotes the stability of the Michaelis complex at the interface. The charge compensation effects on these three residues also account for the differences in the anionic interface preference of the evolutionarily divergent secreted PLA2. PMID- 11015211 TI - Cooperative modulation of protein kinase CK2 by separate domains of its regulatory beta-subunit. AB - Protein kinase CK2 ("casein kinase 2") holoenzyme is composed of two catalytic (alpha and/or alpha') and two regulatory beta-subunits. A truncated form of the beta-subunit lacking its C-terminal region (betaDelta171-215) has lost the ability to stably associate with the catalytic subunits and to display a number of properties which are mediated by structural elements still present in its sequence, notably down-regulation of catalytic activity, autophosphorylation, and responsiveness to polycationic effectors. All these functions are restored by simultaneous addition of a synthetic peptide reproducing the deleted fragment, beta170-215, which is able to associate with the catalytic subunits and to stimulate catalytic activity. This peptide includes a segment displaying significant sequence similarity with a region of cyclin A which interacts with the PSTAIRE motif of CDK2 eliciting its catalytic activity. A peptide reproducing this sequence (beta181-203), but not its derivative in which three nonpolar side chains have been replaced by polar ones, interacts with the alpha-subunit and stimulates its catalytic activity; it also partially restores the ability of truncated betaDelta171-215 to autophosphorylate. These data disclose the essential role of a structural module located between residues 181 and 203 in conferring regulatory properties to the beta-subunit of CK2. PMID- 11015212 TI - Myosin motor domain lever arm rotation is coupled to ATP hydrolysis. AB - We have investigated coupling of lever arm rotation to the ATP binding and hydrolysis steps for the myosin motor domain. In several current hypotheses of the mechanism of force production by muscle, the primary mechanical feature is the rotation of a lever arm that is a subdomain of the myosin motor domain. In these models, the lever arm rotates while the myosin motor domain is free, and then reverses the rotation to produce force while it is bound to actin. These mechanical steps are coupled to steps in the ATP hydrolysis cycle. Our hypothesis is that ATP hydrolysis induces lever arm rotation to produce a more compact motor domain that has stored mechanical energy. Our approach is to use transient electric birefringence techniques to measure changes in hydrodynamic size that result from lever arm rotation when various ligands are bound to isolated skeletal muscle myosin motor domain in solution. Results for ATP and CTP, which do support force production by muscle fibers, are compared to those of ATPgammaS and GTP, which do not. Measurements are also made of conformational changes when the motor domain is bound to NDP's and PP(i) in the absence and presence of the phosphate analogue orthovanadate, to determine the roles the nucleoside moieties of the nucleotides have on lever arm rotation. The results indicate that for the substrates investigated, rotation does not occur upon substrate binding, but is coupled to the NTP hydrolysis step. The data are consistent with a model in which only substrates that produce a motor domain-NDP-P(i) complex as the steady-state intermediate make the motor domain more compact, and only those substrates support force production. PMID- 11015213 TI - Equilibrium folding of dimeric class mu glutathione transferases involves a stable monomeric intermediate. AB - The conformational stabilities of two homodimeric class mu glutathione transferases (GSTM1-1 and GSTM2-2) were studied by urea- and guanidinium chloride induced denaturation. Unfolding is reversible and structural changes were followed with far-ultraviolet circular dichroism, tryptophan fluorescence, enzyme activity, chemical cross-linking, and size-exclusion chromatography. Disruption of secondary structure occurs as a monophasic transition and is independent of protein concentration. Changes in tertiary structure occur as two transitions; the first is protein concentration dependent, while the second is weakly dependent (GSTM1-1) or independent (GSTM2-2). The second transition corresponds with the secondary structure transition. Loss in catalytic activity occurs as two transitions for GSTM1-1 and as one transition for GSTM2-2. These transitions are dependent upon protein concentration. The first deactivation transition coincides with the first tertiary structure transition. Dimer dissociation occurs prior to disruption of secondary structure. The data suggest that the equilibrium unfolding/refolding of the class mu glutathione transferases M1-1 and M2-2 proceed via a three-state process: N(2) <--> 2I <--> 2U. Although GSTM1-1 and GSTM2-2 are homologous (78% identity/94% homology), their N(2) tertiary structures are not identical. Dissociation of the GSTM1-1 dimer to structured monomers (I) occurs at lower denaturant concentrations than for GSTM2-2. The monomeric intermediate for GSTM1-1 is, however, more stable than the intermediate for GSTM2-2. The intermediates are catalytically inactive and display nativelike secondary structure. Guanidinium chloride-induced denaturation yields monomeric intermediates, which have a more loosely packed tertiary structure displaying enhanced solvent exposure of its tryptophans and enhanced ANS binding. The three state model for the class mu enzymes is in contrast to the equilibrium two-state models previously proposed for representatives of classes alpha/pi/Sj26 GSTs. Class mu subunits appear to be intrinsically more stable than those of the other GST classes. PMID- 11015214 TI - Acid denaturation of recombinant porcine growth hormone: formation and self association of folding intermediates. AB - We have investigated the conformational changes incurred during the acid-induced unfolding and self-association of recombinant porcine growth hormone (pGH). Acidification (pH 8 to pH 2) of pGH resulted in intrinsic fluorescence, UV absorbance, and near-UV CD transitions centered at pH 4.10. At pH 2.0, a red shift in the fluorescence emission maximum of approximately 3 nm and a 15% loss of the far-UV CD signal at 222 nm imply that the protein did not become extensively unfolded. Acidification in the presence of 4 M urea resulted in similar pH-dependent transitions. However, these occurred at a higher pH (approximately 5.2). At pH 2.0 + 4 M urea, an 8 nm red shift in the fluorescence emission maximum suggests that unfolding was greater than in the absence of urea. The presence of a prominent peak centered at 298 nm in the near-UV CD spectrum, which is absent without urea, signifies further differences in the intermediates generated at pH 2. Sedimentation equilibrium experiments in the analytical ultracentrifuge showed that native pGH and the partially unfolded intermediates reversibly self-associate. Self-association was strongly promoted at pH 2 while urea reduced self-association at both pH 8 and pH 2. These results demonstrate that acidification of pGH in the absence or presence of 4 M urea induced the formation of molten globule-like states with measurable differences in conformation. Similarities and differences in these structural conformations with respect to other growth hormones are discussed. PMID- 11015215 TI - Cooperative assembly of a nativelike ubiquitin structure through peptide fragment complexation: energetics of peptide association and folding. AB - Peptide fragments corresponding to the N- and C-terminal portions of bovine ubiquitin, U(1-35) and U(36-76), are shown by NMR to associate in solution to form a complex of modest stability (Kassn approximately 1.4 x 10(5) M(-1) at pH 7.0), with NMR features characteristic of a nativelike structure. The complex undergoes cold denaturation, with temperature-dependent estimates of stability from NMR indicating a DeltaC(p) degrees for fragment complexation in good agreement with that determined for native ubiquitin, suggesting that fragment association results in the burial of a similar hydrophobic surface area. The stability of the complex shows appreciable pH dependence, suggesting that ionic interactions on the surface of the protein contribute significantly. However, denaturation studies of native ubiquitin in the presence of guanidine hydrochloride (Gdn.HCl) show little pH dependence, suggesting that ionic interactions may be "screened" by the denaturant, as recently suggested. Examination of the conformation of the isolated peptide fragments has shown evidence for a low population of nativelike structure in the N-terminal beta hairpin (residues 1-17) and weak nascent helical propensity in the helical fragment (residues 21-35). In contrast, the C-terminal peptide (36-76) shows evidence in aqueous solution, from some Halpha chemical shifts, for nonnative phi and psi angles; nonnative alpha-helical structure is readily induced in the presence of organic cosolvents, indicating that tertiary interactions in both native ubiquitin and the folded fragment complex strongly dictate its structural preference. The data suggest that the N-terminal fragment (1-35), where interaction between the helix and hairpin requires the minimum loss of conformational entropy, may provide the nucleation site for fragment complexation. PMID- 11015216 TI - Thermodynamic and structural studies of cavity formation in proteins suggest that loss of packing interactions rather than the hydrophobic effect dominates the observed energetics. AB - The hydrophobic effect is widely believed to be an important determinant of protein stability. However, it is difficult to obtain unambiguous experimental estimates of the contribution of the hydrophobic driving force to the overall free energy of folding. Thermodynamic and structural studies of large to small substitutions in proteins are the most direct method of measuring this contribution. We have substituted the buried residue Phe8 in RNase S with alanine, methionine, and norleucine. Binding thermodynamics and structures were characterized by titration calorimetry and crystallography, respectively. The crystal structures of the RNase S F8A, F8M, and F8Nle mutants indicate that the protein tolerates the changes without any main chain adjustments. The correlation of structural and thermodynamic parameters associated with large to small substitutions was analyzed for nine mutants of RNase S as well as 32 additional cavity-containing mutants of T4 lysozyme, human lysozyme, and barnase. Such substitutions were typically found to result in negligible changes in DeltaC(p)() and positive values of both DeltaDeltaH degrees and DeltaDeltaS of folding. Enthalpic effects were dominant, and the sign of DeltaDeltaS is the opposite of that expected from the hydrophobic effect. Values of DeltaDeltaG degrees and DeltaDeltaH degrees correlated better with changes in packing parameters such as residue depth or occluded surface than with the change in accessible surface area upon folding. These results suggest that the loss of packing interactions rather than the hydrophobic effect is a dominant contributor to the observed energetics for large to small substitutions. Hence, estimates of the magnitude of the hydrophobic driving force derived from earlier mutational studies are likely to be significantly in excess of the actual value. PMID- 11015217 TI - Contribution of salt bridges near the surface of a protein to the conformational stability. AB - Salt bridges play important roles in the conformational stability of proteins. However, the effect of a surface salt bridge on the stability remains controversial even today; some reports have shown little contribution of a surface salt bridge to stability, whereas others have shown a favorable contribution. In this study, to elucidate the net contribution of a surface salt bridge to the conformational stability of a protein, systematic mutant human lysozymes, containing one Glu to Gln (E7Q) and five Asp to Asn mutations (D18N, D49N, D67N, D102N, and D120N) at residues where a salt bridge is formed near the surface in the wild-type structure, were examined. The thermodynamic parameters for denaturation between pH 2.0 and 4.8 were determined by use of a differential scanning calorimeter, and the crystal structures were analyzed by X-ray crystallography. The denaturation Gibbs energy (DeltaG) of all mutant proteins was lower than that of the wild-type protein at pH 4, whereas there was little difference between them near pH 2. This is caused by the fact that the Glu and Asp residues are ionized at pH 4 but protonated at pH 2, indicating a favorable contribution of salt bridges to the wild-type structure at pH 4. Each contribution was not equivalent, but we found that the contributions correlate with the solvent inaccessibility of the salt bridges; the salt bridge contribution was small when 100% accessible, while it was about 9 kJ/mol if 100% inaccessible. This conclusion indicates how to reconcile a number of conflicting reports about role of surface salt bridges in protein stability. Furthermore, the effect of salts on surface salt bridges was also examined. In the presence of 0.2 M KCl, the stability at pH 4 decreased, and the differences in stability between the wild-type and mutant proteins were smaller than those in the absence of salts, indicating the compensation to the contribution of salt bridges with salts. Salt bridges with more than 50% accessibility did not contribute to the stability in the presence of 0.2 M KCl. PMID- 11015218 TI - The residual pro-part of cathepsin C fulfills the criteria required for an intramolecular chaperone in folding and stabilizing the human proenzyme. AB - The 13.5 kDa N-terminal part of the propeptide remains associated with mature cathepsin C after proteolytic activation and excision of the activation peptide. This residual pro-part, isolated from the recombinant enzyme, folds spontaneously and rapidly to a stable, compact monomer with secondary structure and stable tertiary interactions. Folding and unfolding kinetics of the residual pro-part with intact disulfides are complex, and accumulation of transient intermediates is observed. The cleaved form of the pro-part isolated from natural human cathepsin C also folds, suggesting that the intact form comprises two folding domains. The linkages of the two disulfide bridges have been established as 30 118 and 54-136 for the native enzyme. The native disulfide bonds can be re-formed from the fully reduced and denatured state by oxidative refolding, resulting in a domain that is spectroscopically indistinguishable from the original refolded residual pro-part. Both disulfides are solvent-exposed and can be reduced in the absence of denaturant. The reduced form retains most or all of the native tertiary structure and is only approximately 2 kcal.mol(-1) less stable than the oxidized form. It folds fast relative to the rate of biosynthesis, to the same conformation as the oxidized form. Folding and disulfide formation are sequential. These results indicate that the proenzyme folds sequentially in vivo and that the residual pro-part constitutes a rapidly and independently folding domain that stabilizes the mature enzyme. It thus fulfills the criteria required of an intramolecular chaperone. It may also be involved in stabilizing the tetrameric structure of the mature enzyme. PMID- 11015219 TI - Mechanism of cyanamide hydration catalyzed by carbonic anhydrase II suggested by cryogenic X-ray diffraction. AB - The three-dimensional structure of a possible intermediate in the hydration reaction of cyanamide to urea catalyzed by human carbonic anhydrase II (hCAII) has been determined by cryocrystallographic techniques. The crystal structure shows that two different adducts are formed under the experimental conditions and that they have different occupancy in the crystal. The high occupancy form consists of a binary hCAII-cyanamide complex where the substrate has replaced the zinc-bound hydroxide anion present in the native enzyme, maintaining the tetrahedral geometry around the metal ion. The second, low-occupancy form consists of a hCAII-cyanamide-water ternary complex where the catalytic zinc ion, still being bound to cyanamide, is approached by a water molecule in a five coordinate adduct. While the first form can be considered a nonproductive complex, the second form may represent an intermediate state of the catalyzed reaction where the water molecule is about to perform a nucleophilic attack on the zinc-activated cyanamide substrate. The structural evidence is consistent with the kinetic data previously reported about this recently described hydrolytic reaction catalyzed by hCAII, and indicates that a different mechanism with respect to that generally accepted for the physiologic carbon dioxide hydration reaction may be adopted by the enzyme, depending on the substrate chemical properties. PMID- 11015220 TI - Role of metals in the reaction catalyzed by protein farnesyltransferase. AB - Protein farnesyltransferase catalyzes the posttranslational farnesylation of several proteins involved in signal transduction, including Ras, and is a target enzyme for antitumor therapies. Efficient product formation catalyzed by protein farnesyltransferase requires an enzyme-bound zinc cation and high concentrations of magnesium ions. In this work, we have measured the pH dependence of the chemical step of product formation, determined under single-turnover conditions, and have demonstrated that the prenylation rate constant is enhanced by two deprotonations. Substitution of the active site zinc by cadmium demonstrated that one of the ionizations reflects deprotonation of the metal-coordinated thiol of the peptide "CaaX" motif, pK(a1) = 6.0. These data provide additional evidence for the direct involvement of a metal-coordinated sulfur nucleophile in catalysis. The second ionization was assigned to a hydroxyl on the pyrophosphate moiety of farnesyl pyrophosphate, pK(a2) = 7.4. Deprotonation of this group is important for binding of magnesium. This second ionization is not observed for catalysis in the absence of magnesium or when the substrate is farnesyl monophosphate. These data indicate that the maximal rate constant for prenylation requires formation of a zinc-coordinated thiolate nucleophile and enhancement of the electrophilic character at C1 of the farnesyl chain by magnesium ion coordination of the pyrophosphate leaving group. PMID- 11015221 TI - Mass spectral characterization of lipooligosaccharides from Haemophilus influenzae 2019. AB - Lipooligosaccharide (LOS) glycoforms from Haemophilus influenzae 2019 were profiled using the high-resolution and accurate mass capabilities of Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Sequence and linkage for two previously unknown LOS glycoforms were subsequently obtained through MSn analyses on FT-ICR and quadrupole ion trap (qIT) instruments. MSn analysis of negative ion precursors confirmed structural details within the lipid moiety, while CID spectra of sodiated precursor ions provided monosaccharide sequence and linkage for the oligosaccharide portion of the molecule. Results obtained in this study indicate that extensive heterogeneity exists within the oligosaccharide moieties in LOS from H. influenzae 2019. More importantly, the data suggest that additional hexose moieties, which are added onto the LOS, are not simple extensions of one particular core structure but rather that structural isomers with different connectivities are present within the heterogeneous mixture. PMID- 11015222 TI - Inositol-1-phosphate synthase from Archaeoglobus fulgidus is a class II aldolase. AB - A gene putatively identified as the Archaeoglobus fulgidus inositol-1-phosphate synthase (IPS) gene was overexpressed to high level (about 30-40% of total soluble cellular proteins) in Escherichia coli. The recombinant protein was purified to homogeneity by heat treatment followed by two column chromatographic steps. The native enzyme was a tetramer of 168 +/- 4 kDa (subunit molecular mass of 44 kDa). At 90 degrees C the K(m) values for glucose-6-phosphate and NAD(+) were estimated as 0.12 +/- 0.04 mM and 5.1 +/- 0.9 microM, respectively. Use of (D)-[5-(13)C]glucose-6-phosphate as a substrate confirmed that the stereochemistry of the product of the IPS reaction was L-myo-inositol-1 phosphate. This archaeal enzyme, with the highest activity at its optimum growth temperature among all IPS reported (k(cat) = 9.6 +/- 0.4 s(-1) with an estimated activation energy of 69 kJ/mol), was extremely heat stable. However, the most unique feature of A. fulgidus IPS was that it absolutely required divalent metal ions for activity. Zn(2+) and Mn(2+) were the best activators with K(D) approximately 1 microM, while NH(4)(+) (a critical activator for all the other characterized IPS enzymes) had no effect on the enzyme. These properties suggested that this archaeal IPS was a class II aldolase. In support of this, stoichiometric reduction of NAD(+) to NADH could be followed spectrophotometrically when EDTA was present along with glucose-6-phosphate. PMID- 11015223 TI - Resonance Raman studies indicate a unique heme active site in prostaglandin H synthase. AB - Prostaglandin H synthase isoforms 1 and 2 (PGHS-1 and -2) catalyze the first two steps in the biosynthesis of prostaglandins. Resonance Raman spectroscopy was used to characterize the PGHS heme active site and its immediate environment. Ferric PGHS-1 has a predominant six-coordinate high-spin heme at room temperature, with water as the sixth ligand. The proximal histidine ligand (or the distal water ligand) of this hexacoordinate high-spin heme species was reversibly photolabile, leading to a pentacoordinate high-spin ferric heme iron. Ferrous PGHS-1 has a single species of five-coordinate high-spin heme, as evident from nu(2) at 1558 cm(-1) and nu(3) at 1471 cm(-1). nu(4) at 1359 cm(-1) indicates that histidine is the proximal ligand. A weak band at 226-228 cm(-1) was tentatively assigned as the Fe-His stretching vibration. Cyanoferric PGHS-1 exhibited a nu(Fe)(-)(CN) line at 446 cm(-1) and delta(Fe)(-)(C)(-)(N) at 410 cm( 1), indicating a "linear" Fe-C-N binding conformation with the proximal histidine. This linkage agrees well with the open distal heme pocket in PGHS-1. The ferrous PGHS-1 CO complex exhibited three important marker lines: nu(Fe)( )(CO) (531 cm(-1)), delta(Fe)(-)(C)(-)(O) (567 cm(-1)), and nu(C)(-)(O) (1954 cm( 1)). No hydrogen bonding was detected for the heme-bound CO in PGHS-1. These frequencies markedly deviated from the nu(Fe)(-)(CO)/nu(C)(-)(O) correlation curve for heme proteins and porphyrins with a proximal histidine or imidazolate, suggesting an extremely weak bond between the heme iron and the proximal histidine in PGHS-1. At alkaline pH, PGHS-1 is converted to a second CO binding conformation (nu(Fe)(-)(CO): 496 cm(-1)) where disruption of the hydrogen bonding interactions to the proximal histidine may occur. PMID- 11015225 TI - Spectroscopic studies of zinc(II)- and cobalt(II)-associated Escherichia coli formamidopyrimidine-DNA glycosylase: extended X-ray absorption fine structure evidence for a metal-binding domain. AB - Formamidopyrimidine-DNA glycosylase (Fpg) is a 30.2 kDa protein that plays an important role in the base excision repair of oxidatively damaged DNA in Escherichia coli. Sequence analysis and genetic evidence suggest that zinc is associated with a C4-type motif, C(244)-X(2)-C(247)-X(16)-C(264)-X(2)-C(267), located at the C-terminus of the protein. The zinc-associated motif has been shown to be essential for damaged DNA recognition. Extended X-ray absorption fine structure (EXAFS) spectra collected on the zinc-associated protein (ZnFpg) in the lyophilized state and in 10% frozen aqueous glycerol solution show directly that the metal is coordinated to the sulfur atom of four cysteine residues. The average Zn-S bond length is 2.33 +/- 0.01 and 2.34 +/- 0.01 A, respectively, in the lyophilized state and in 10% frozen aqueous glycerol solution. Fpg was also expressed in minimal medium supplemented with cobalt nitrate to yield a blue colored protein that was primarily cobalt-associated (CoFpg). The profiles of the circular dichroism spectra for CoFpg and ZnFpg are identical, suggesting that the substitution of Co(2+) for Zn(2+) does not alter the structure of Fpg. A similar conclusion is reached upon the analysis of two-dimensional (15)N/(1)H HSQC spectra of uniformly (15)N-labeled samples of ZnFpg and CoFpg; the spectra are similar and display features characteristic of a structured protein. Biochemical assays with a 54 nt DNA oligomer containing 7, 8-dihydro-8-oxoguanine at a specific location show that CoFpg and ZnFpg are equally active at cleaving the DNA at the site of the oxidized guanine. EXAFS spectra of CoFpg indicate that the cobalt is coordinated to the sulfur atom of four cysteine residues with an average Co-S bond length of 2.28 +/- 0.01 and 2.29 +/- 0.01 A, respectively, in the lyophilized state and in 10% frozen aqueous glycerol solution. The structural similarity between CoFpg and ZnFpg suggests that it is biologically relevant to use the paramagnetic properties of Co(2+) as a structural probe. PMID- 11015224 TI - UreE stimulation of GTP-dependent urease activation in the UreD-UreF-UreG-urease apoprotein complex. AB - The activation of metal-containing enzymes often requires the participation of accessory proteins whose roles are poorly understood. In the case of Klebsiella aerogenes urease, a nickel-containing enzyme, metallocenter assembly requires UreD, UreF, and UreG acting as a protein chaperone complex and UreE serving as a nickel metallochaperone. Urease apoprotein within the UreD-UreF-UreG-urease apoprotein complex is activated to wild-type enzyme activity levels under physiologically relevant conditions (100 microM bicarbonate and 20 microM Ni2+) in a process that requires GTP and UreE. The GTP concentration needed for optimal activation is greatly reduced in the presence of UreE compared to that required in its absence. The amount of UreE provided is critical, with maximal activation observed at a concentration equal to that of Ni2+. On the basis of its ability to facilitate urease activation in the presence of chelators, UreE is proposed to play an active role in transferring Ni2+ to urease apoprotein. Studies involving site-directed variants of UreE provide evidence that His96 has a direct role in metal transfer. The results presented here parallel those obtained from previous in vivo studies, demonstrating the relevance of this in vitro system to the cellular metallocenter assembly process. PMID- 11015226 TI - Proteolytic activation of recombinant pro-memapsin 2 (pro-beta-secretase) studied with new fluorogenic substrates. AB - Memapsin 2 (beta-secretase), a membrane-anchored aspartic protease, is involved in the cleavage of beta-amyloid precursor protein to form beta-amyloid peptide. The primary structure of memapsin 2 suggests that it is synthesized in vivo as pro-memapsin 2 and converted to memapsin 2 by an activating protease [Lin et al. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 1456-1460]. To simulate this activation mechanism and to produce stable mature memapsin 2 for kinetic/specificity studies, we have investigated the activation of recombinant pro-memapsin 2 by several proteases with trypsin-like specificity. Clostripain, kallikrein, and trypsin increased the activity of pro-memapsin 2. Clostripain activation was accompanied by the cleavage of the pro region to form mainly two activation products, Leu(30p)- and Gly(45p)-memapsin 2. Another activation product, Leu(28p) memapsin 2, was also purified. Kinetics of the activated memapsin 2 were compared with pro-memapsin 2 using two new fluorogenic substrates, Arg-Glu(5-[(2 aminoethyl)amino]naphthalene-1-sulfonic acid (EDANS))-Glu-Val-Asn-Leu-Asp-Ala-Glu Phe-Lys(4-(4-dimethylaminophe nyl azo)benzoic acid (DABCYL))-Arg and (7 methoxycoumarin-4-yl)acetyl (MCA))-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys(2,4 dinitrophenyl (DNP)). These results establish that the activity of pro-memapsin 2 stems from a part-time and reversible uncovering of its active site by its pro region. Proteolytic removal of part of the pro-peptide at Leu(28p) or Gly(45p), which diminishes the affinity of the shortened pro-peptide to the active site, results in activated memapsin 2. These results also suggest that Glu(33p) memapsin 2 observed in the cells expressing this enzyme [Vassar et al. (1999) Science 286, 735-741; Yan et al. (1999) Nature 402, 533-537] is an active intermediate of in vivo activation, or that the peptide Glu(33p)-Arg(44p) may serve a regulatory role. PMID- 11015227 TI - "Paper-clip" type triple helix formation by 5'-d-(TC)3Ta(CT)3Cb(AG)3 (a and b = 0 4) as a function of loop size with and without the pseudoisocytosine base in the Hoogsteen strand. AB - The formation of a DNA "paper-clip" type triple helix (triplex) with a common sequence 5'-d-(TC)(3)T(a)()(CT)(3)C(b)()(AG)(3) (a and b = 0-4) was studied by UV thermal melting experiments and CD spectra. These DNA oligomers form triplexes and duplexes under slightly acidic and neutral conditions, respectively. The stability of the formed triplexes (at pH 4.5) or duplexes (at pH 7.0 or 8.0) does not vary significantly with the size of the loops (a and b = 1-4). At pH 6.0, the triplex stability is, however, a function of a and b. It is also interesting to note that the oligomer 5'-d-(TC)(3)(CT)(3)(AG)(3) (a and b = 0) forms a stable triplex at pH 4.5 with a slightly lower T(m) value, due to dissociation of a base triad at one end and a distorted base triad at the other, observed by (1)H NMR. Thus, we have here a model system, 5'-d-(TC)(3)T(a)(CT)(3)C(b)(AG)(3), that could form a triplex effectively with (a and b = 1-4) and without (a and b = 0) loops under acidic conditions. In addition, the triplex formation of oligomers with replacement of one, two, or three 2'-deoxycytidine in the Hoogsteen strand by either 2'-deoxypseudoisocytidine (D) or 2'-O-methylpseudoisocytidine (M) was also studied in the sequence 5'-d-(TX)(3)T(2)(CT)(3)C(2)(AG)(3) (where X is C, D, or M). Both CD spectra and UV melting results showed that only D3 [(TX)(3) = (TD)(3)] and M3 [(TX)(3) = (TM)(3)] were able to form the paper-clip structure under both neutral and acidic conditions. This is because the N(3)H of a pseudoisocytosine base can serve as a proton donor without protonation. We hereby proved that the 2'-deoxypseudoisocytidine, similar to 2'-O methylpseudoisocytidine, could replace 2'-deoxycytidine in the Hoogsteen strand to provide triplex formation at neutral pH. PMID- 11015228 TI - Multiple folding pathways for the P4-P6 RNA domain. AB - We recently described site-specific pyrene labeling of RNA to monitor Mg(2+) dependent equilibrium formation of tertiary structure. Here we extend these studies to follow the folding kinetics of the 160-nucleotide P4-P6 domain of the Tetrahymena group I intron RNA, using stopped-flow fluorescence with approximately 1 ms time resolution. Pyrene-labeled P4-P6 was prepared using a new phosphoramidite that allows high-yield automated synthesis of oligoribonucleotides with pyrene incorporated at a specific 2'-amino-2' deoxyuridine residue. P4-P6 forms its higher-order tertiary structure rapidly, with k(obs) = 15-31 s(-1) (t(1/2) approximately 20-50 ms) at 35 degrees C and [Mg(2+)] approximately 10 mM in Tris-borate (TB) buffer. The folding rate increases strongly with temperature from 4 to 45 degrees C, demonstrating a large activation enthalpy DeltaH(double dagger) approximately 26 kcal/mol; the activation entropy DeltaS(double dagger) is large and positive. In low ionic strength 10 mM sodium cacodylate buffer at 35 degrees C, a slow (t(1/2) approximately 1 s) folding component is also observed. The folding kinetics are both ionic strength- and temperature-dependent; the slow phase vanishes upon increasing [Na(+)] in the cacodylate buffer, and the kinetics switch completely from fast at 30 degrees C to slow at 40 degrees C. Using synchrotron hydroxyl radical footprinting, we confirm that fluorescence monitors the same kinetic events as hydroxyl radical cleavage, and we show that the previously reported slow P4-P6 folding kinetics apply only to low ionic strength conditions. One model to explain the fast and slow folding kinetics postulates that some tertiary interactions are present even without Mg(2+) in the initial state. The fast kinetic phase reflects folding that is facilitated by these interactions, whereas the slow kinetics are observed when these interactions are disrupted at lower ionic strength and higher temperature. PMID- 11015229 TI - Spin-label electron spin resonance studies on the mode of anchoring and vertical location of the N-acyl chain in N-acylphosphatidylethanolamines. AB - Electron spin resonance (ESR) studies have been performed on N-myristoyl dimyristoylphosphatidylethanolamine (N-14-DMPE) membranes using both phosphatidylcholines spin-labeled at different positions in the sn-2 acyl chain and N-acyl phosphatidylethanolamines spin-labeled in the N-acyl chain to characterize the location and mobility of the N-acyl chain in the lipid membranes. Comparison of the positional dependences of the spectral data for the two series of spin-labeled lipids suggests that the N-acyl chain is positioned at approximately the same level as the sn-2 chain of the phosphatidylcholine spin label. Further, similar conclusions are reached when the ESR spectra of the N acyl PE spin-labels in dimyristoylphosphatidylcholine (DMPC) or dimyristoylphosphatidylethanolamine (DMPE) host matrixes are compared with those of phosphatidylcholine spin-labels in these two lipids. Finally, the chain ordering effect of cholesterol has also been found to be similar for the N-acyl PE spin-label and PC spin-labels, when the host matrix is either DMPC and cholesterol or N-14-DMPE and cholesterol at a 6:4 mole ratio. In both cases, the gel-to-liquid crystalline phase transition is completely abolished but cholesterol perturbs the gel-phase mobility of N-14-DMPE more readily than that of DMPC. These results demonstrate that the long N-acyl chains are anchored firmly in the hydrophobic interior of the membrane, in an orientation that is parallel to that of the O-acyl chains, and are located at nearly the same vertical position as that of the sn-2 acyl chains in the lipid bilayer. There is a high degree of dynamic compatibility between the N-acyl chains and the O-acyl chains of the lipid bilayer core, although bilayers of N-acyl phosphatidylethanolamines possess a more hydrophobic interior than phosphatidylcholine bilayers. These results provide a structural basis for rationalizing the biological properties of NAPEs. PMID- 11015230 TI - Dynamin is membrane-active: lipid insertion is induced by phosphoinositides and phosphatidic acid. AB - Dynamin is a large GTPase involved in the regulation of membrane constriction and fission during receptor-mediated endocytosis. Dynamin contains a pleckstrin homology domain which is essential for endocytosis and which binds to anionic phospholipids. Here, we show for the first time that dynamin is a membrane-active molecule capable of penetrating into the acyl chain region of membrane lipids. Lipid penetration is strongly stimulated by phosphatidic acid (PA), phosphatidylinositol 4-phosphate, and phosphatidylinositol 4, 5-bisphosphate. Though binding is more efficient in the presence of the phosphoinositides, a much larger part of the dynamin molecule penetrates into PA-containing mixed-lipid systems. Thus, local lipid metabolism will dramatically influence dynamin-lipid interactions, and dynamin-lipid interactions are likely to play an important role in dynamin-dependent endocytosis. Our data suggest that dynamin is directly involved in membrane destabilization, a prerequisite to membrane fission. PMID- 11015231 TI - Striking stabilization of Arc repressor by an engineered disulfide bond. AB - A solvent-exposed Cys11-Cys11' disulfide bond was designed to link the antiparallel strands of the beta sheet both in the Arc repressor dimer and in a single-chain variant in which the Arc subunits are connected by a 15-residue peptide tether. In both proteins, the presence of the disulfide bond increased the T(m) by approximately 40 degrees C. In the single-chain background, the disulfide bond stabilized Arc by 8.5 kcal/mol relative to the reduced form, a significantly larger degree of stabilization than caused by other engineered disulfides and most natural disulfides. This exceptional stabilization arises from a modest effective concentration of the Cys11-Cys11' disulfide in the native state (71 M) and an anomalously low effective concentration in the denatured state (40 microM). Disulfide cross-linking of the two beta strands in the single chain Arc background accelerated refolding by a factor of 170 into the sub microsecond time scale. However, the major energetic effect of the disulfide occurs after the transition state for Arc refolding, slowing unfolding by 200 000 fold. PMID- 11015233 TI - Biophysical characterization of elongin C from saccharomyces cerevisiae PMID- 11015232 TI - Reaction of hydrogen peroxide with ferrylhemoglobin: superoxide production and heme degradation. AB - The reaction of Fe(II) hemoglobin (Hb) but not Fe(III) hemoglobin (metHb) with hydrogen peroxide results in degradation of the heme moiety. The observation that heme degradation was inhibited by compounds, which react with ferrylHb such as sodium sulfide, and peroxidase substrates (ABTS and o-dianisidine), demonstrates that ferrylHb formation is required for heme degradation. A reaction involving hydrogen peroxide and ferrylHb was demonstrated by the finding that heme degradation was inihibited by the addition of catalase which removed hydrogen peroxide even after the maximal level of ferrylHb was reached. The reaction of hydrogen peroxide with ferrylHb to produce heme degradation products was shown by electron paramagnetic resonance to involve the one-electron oxidation of hydrogen peroxide to the oxygen free radical, superoxide. The inhibition by sodium sulfide of both superoxide production and the formation of fluorescent heme degradation products links superoxide production with heme degradation. The inability to produce heme degradation products by the reaction of metHb with hydrogen peroxide was explained by the fact that hydrogen peroxide reacting with oxoferrylHb undergoes a two-electron oxidation, producing oxygen instead of superoxide. This reaction does not produce heme degradation, but is responsible for the catalytic removal of hydrogen peroxide. The rapid consumption of hydrogen peroxide as a result of the metHb formed as an intermediate during the reaction of reduced hemoglobin with hydrogen peroxide was shown to limit the extent of heme degradation. PMID- 11015235 TI - Detection of circulating lipopolysaccharide-bound monocytes in children with gram negative sepsis. AB - The possibility that gram-negative sepsis can be diagnosed by detection of lipopolysaccharide (LPS) bound on the surface of monocytes in the circulation of patients with gram-negative sepsis was investigated. Peripheral monocytes were analyzed by flow cytometer and an anti-LPS monoclonal antibody in 3 groups: children with gram-negative sepsis, children with gram-positive sepsis, and healthy children. LPS-bound monocytes were found in all patients with gram negative sepsis but not in children with gram-positive sepsis or in healthy children. Therefore, the flow cytometry method developed for this study may be a novel method for diagnosing gram-negative sepsis. PMID- 11015234 TI - Human immune response to streptococcal inhibitor of complement, a serotype M1 group A Streptococcus extracellular protein involved in epidemics. AB - Streptococcal inhibitor of complement (Sic) is a highly polymorphic extracellular protein made by serotype M1 group A Streptococcus strains that contributes to bacterial persistence in the mammalian upper respiratory tract. New variants of the Sic protein arise very rapidly by positive selection in human populations during M1 epidemics. The human antibody response to Sic was analyzed. Of 636 persons living in diverse localities, 43% had anti-Sic serum antibodies, but only 16.4% had anti-M1 protein serum antibody. Anti-Sic antibody was also present in nasal wash specimens in high frequency. Linear B cell epitope mapping showed that serum antibodies recognized epitopes located in structurally variable regions of Sic and the amino terminal hypervariable region of the M1 protein. Phage display analyses confirmed that the polymorphic regions of Sic are primary targets of host antibodies. These results support the hypothesis that selection of Sic variants occurs on mucosal surfaces by a mechanism that involves acquired host antibody. PMID- 11015236 TI - Inverse relationship between gastric colonization of Helicobacter pylori and diarrheal illnesses in children: results of a population-based cross-sectional study. AB - It has been suggested that carriage of Helicobacter pylori may protect against infections by exogenous intestinal pathogens. An analysis was done of all children who were screened for school fitness during 1996-1998 in Ulm, Germany, to compare rates of diarrheal illnesses in H. pylori-positive and H. pylori negative children. Of 2477 5-8-year-old children studied, 304 (12.3%) were H. pylori-positive by carbon 13-labeled urea breath test. For H. pylori-positive children, diarrhea within the prior 3 months was less often reported than for H. pylori-negative children (54.3% vs. 76.1%; P<.001, adjusted for nationality). Compared with H. pylori-negative children, the odds ratio (OR) for the occurrence of diarrhea within the prior 3 months was 0.37 (95% confidence interval [CI], 0.28-0.49) for H. pylori-positive children; after adjustment for covariates, the OR was 0.56 (95% CI, 0.42-0.76). These data support the hypothesis that H. pylori colonization may protect against diarrheagenic gastrointestinal infections. PMID- 11015237 TI - Two-thumb vs. two-finger chest compression in an infant model of prolonged cardiopulmonary resuscitation. AB - OBJECTIVE: Previous experiments in the authors' swine lab have shown that cardiopulmonary resuscitation (CPR) using two-thumb chest compression with a thoracic squeeze (TT) produces higher blood and perfusion pressures when compared with the American Heart Association (AHA)-recommended two-finger (TF) technique. Previous studies were of short duration (1-2 minutes). The hypothesis was that TT would be superior to TF during prolonged CPR in an infant model. METHODS: This was a prospective, randomized crossover experiment in a laboratory setting. Twenty-one AHA-certified rescuers performed basic CPR for two 10-minute periods, one with TT and the other with TF. Trials were separated by 2-14 days, and the order was randomly assigned. The experimental circuit consisted of a modified manikin with a fixed-volume arterial system attached to a neonatal monitor via an arterial pressure transducer. The arterial circuit was composed of a 50-mL bag of normal saline solution (air removed) attached to the manikin chest plate and connected to the transducer with a 20-gauge intravenous catheter and tubing. Rescuers were blinded to the arterial pressure tracing. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) were recorded in mm Hg, and pulse pressures (PPs) were calculated. Data were analyzed with two-way repeated-measures analysis of variance. Sphericity assumed modeling, with Greenhouse-Geisser and Huynh-Feldt adjustments, was applied. RESULTS: Marginal means for TT SBP (68.9), DBP (17.6), MAP (35.3), and PP (51.4) were higher than for TF SBP (44.8), DBP (12.5), MAP (23.3), and PP (32.2). All four pressures were significantly different between the two techniques (p< or =0.001). CONCLUSION: In this infant CPR model, TT chest compression produced higher MAP, SBP, DBP, and PP when compared with TF chest compression during a clinically relevant duration of prolonged CPR. PMID- 11015238 TI - Development of a histomorphologic scale to quantify cutaneous scars after burns. AB - OBJECTIVE: Cutaneous wound healing in adults invariably results in scarring; however, there are few scales to quantify the degree of such scarring. The authors developed a histomorphologic scale for quantifying scarring after cutaneous burn injury. METHODS: As part of a randomized trial comparing a variety of burn therapies, 40 partial-thickness burns were created on the backs and flanks of anesthetized pigs and treated with a tissue adhesive, antibiotic ointment, occlusive dressing, or dry gauze. Gross scar appearance was independently assessed by two investigators at 90 days on a 100-mm visual analog scale (VAS) marked "best appearance" at the high end. One of the investigators repeated the observation 30 days later. Full-thickness biopsies were taken 90 days after injury and evaluated histologically by a dermatopathologist for the presence of hyperkeratosis, epidermal hyperplasia, presence and depth of scar (defined as abnormally oriented collagen under polarized light), fibroplasia, vascular proliferation, and absence of adnexa, including hair follicles, apocrine glands, and smooth muscles. One point was assigned for each category in the presence of a normal finding, whereas an abnormal finding was assigned a score of zero. The normal dermis (absence of abnormal collagen) was given a score of 3, while decreasing scores of 2 to 0 were given for progressively deeper scars (i.e., 2 for papillary dermis, 1 for upper half of reticular dermis, and 0 for deep dermal lower half). The total histomorphologic score was derived by adding the scores on the individual items. The score ranges from 0 to 10 from worst scarring to absence of scarring, respectively. A subset of observations was evaluated a second time by one of the observers one month later. Intraobserver reliability of the histomorphologic scale was assessed with Spearman's correlation. Inter- and intraobserver Pearson's correlations for the gross scar VAS were calculated, and the correlation between gross and histomorphologic scores was assessed. RESULTS: Intraobserver correlation for individual histomorphologic categories ranged from 0.19 to 1.00. Intraobserver correlation for the total histologic score was 0.95. Inter- and intraobserver correlations for the gross scar VAS were 0.8 each. Correlation between the histomorphologic scale and the gross scar VAS was 0.38. CONCLUSIONS: A new reliable histomorphologic method for quantifying and scoring cutaneous scars is described together with a reliable scar VAS. However, these two scales are not highly correlated. PMID- 11015239 TI - Calcium and digoxin vs. calcium alone for severe verapamil toxicity. AB - Calcium chloride (CaCl(2)) is ineffective in severe calcium channel antagonist overdoses. Digoxin increases intracellular calcium by inhibiting the sodium potassium adenosine triphosphatase enzymes. OBJECTIVE: To examine the effect of calcium and digoxin on the treatment of verapamil toxicity. METHODS: Sixteen dogs were instrumented to monitor hemodynamics. Verapamil toxicity (50% decrease in mean arterial pressure) was induced with verapamil (VER) at 6 mg/kg/hr and maintained for 30 minutes by titrating the VER rate. Following toxicity, the dogs received either digoxin (0.018 mg/kg) (DIG) (n = 8) or saline (No-DIG) (n = 8). Both groups received VER at three sequential rates (1 mg/kg/hr from 0 to 90 min, 6 mg/kg/hr from 90 to 130 min, and 18 mg/kg/hr from 130 to 170 min). Calcium boluses were given (500 mg at 0 and 15 min; 1 g at 140, 150, and 160 min). Data were analyzed using a repeated-measures analysis of covariance comparing DIG vs No-DIG across the infusion rates and time. Animal weight, does of VER administered during the toxicity phase, and baseline values were included as covariates. Mortality rates were compared at 230 minutes following a total dose of 500 mg of VER. RESULTS: The DIG group had a higher systolic blood pressure (SBP) than the No-DIG group during the 1-mg/kg/hr (early p = 0.028, late p = 0.01), 6-mg/kg/hr (p = 0.051), and 18-mg/kg/hr (p = 0.038) VER infusion rates. There were no deaths in the DIG group and four deaths in the No-DIG group (Fisher = 0.08). Neither ventricular tachycardia nor ventricular fibrillation developed in either group. Other hemodynamic parameters did not show significant changes. CONCLUSIONS: In a model of severe verapamil toxicity, digoxin plus calcium raised SBP and did not result in ventricular arrhythmias when compared with calcium alone. PMID- 11015240 TI - The efficacy of nebulized racemic epinephrine in children with acute asthma: a randomized, double-blind trial. AB - OBJECTIVES: Recent work in bronchiolitis has demonstrated a significant clinical improvement in children treated with epinephrine over nebulized salbutamol. The objective of this study was to determine whether nebulized epinephrine, as compared with nebulized salbutamol, causes a greater clinical improvement in children with acute asthma. METHODS: Children, aged 1 to 17 years, with acute asthma presenting to the emergency department (ED) were eligible. In this double blind study, patients were randomly allocated to receive either salbutamol or racemic epinephrine by nebulization at 0, 20, and 40 minutes. All patients received oral steroids. The primary outcome measure was a change in pulmonary index score (PIS). RESULTS: One hundred twenty patients were randomized. The groups were comparable in terms of age, gender, asthma severity, previous treatments, and use of inhaled steroids. There was no significant difference between treatments in the change in PIS, length of stay, admission to hospital, or relapse rate. The epinephrine-treated group had significantly more minor side effects (such as excess or brownish nasal discharge). CONCLUSIONS: There is no significant clinical benefit of nebulized epinephrine over salbutamol in children 1-17 years old with mild to moderate acute asthma. Salbutamol remains the treatment of choice in children with known asthma. PMID- 11015241 TI - Concordance of clinical findings and clinical judgment in the diagnosis of streptococcal pharyngitis. AB - OBJECTIVE: It is uncertain how reliably clinicians apply clinical predictors of group A beta hemolytic streptococcal pharyngitis (GABHSP) to form a clinical impression, and how reliably this impression predicts culture results. The objective was to study clinician accuracy in diagnosing GABHSP. METHODS: This was a prospective cohort study, conducted at an urgent care center of a major university. A convenience sample of 218 patients, aged 9-83 years, presenting with sore throat, was enrolled. Symptoms and signs of pharyngitis were documented on a standardized form; the likelihood of GABHSP was plotted on a visual analog scale; and throat culture was obtained. A comparison was then made between the clinical impression on presentation and the throat culture result. RESULTS: Throat cultures were positive for GABHSP in 41 patients (19%). The probability of GABHSP was related to node size and tenderness, tonsillar exudate and hypertrophy, and pharyngeal erythema (p<0.05); but not throat soreness, degree of fever, or cough. A strong clinical impression of GABHSP (>50% on the visual analog scale) was associated with tonsillar exudate and hypertrophy, tender nodes, and pharyngeal erythema. Together, these four predictors had a sensitivity of 71%, a specificity of 77%, and a positive predictive value of 46%. CONCLUSIONS: Clinicians in this study based their impression of GABHSP on the most reliable symptoms and signs. While a strong clinical suspicion of GABHSP predicted a greater probability of positive culture, the clinicians consistently overestimated the probability of GABHSP. Symptoms and signs predict GABHSP unreliably when used alone; they are helpful in modifying estimates of disease probability to facilitate optimal use of laboratory tests and antibiotics. PMID- 11015242 TI - Early discharge of patients with presumed opioid overdose: development of a clinical prediction rule. AB - OBJECTIVE: To develop a clinical prediction rule to identify patients who can be safely discharged one hour after the administration of naloxone for presumed opioid overdose. METHODS: Patients who received naloxone for known or presumed opioid overdose were formally evaluated one hour later for multiple potential predictor variables. Patients were classified into two groups: those with adverse events within 24 hours and those without. Using classification and regression tree methodology, a decision rule was developed to predict safe discharge. RESULTS: Clinical findings from 573 patients allowed us to develop a clinical prediction rule with a sensitivity of 99% (95% CI = 96% to 100%) and a specificity of 40% (95% CI = 36% to 45%). Patients with presumed opioid overdose can be safely discharged one hour after naloxone administration if they: 1) can mobilize as usual; 2) have oxygen saturation on room air of >92%; 3) have a respiratory rate >10 breaths/min and <20 breaths/min; 4) have a temperature of >35.0 degrees C and <37.5 degrees C; 5) have a heart rate >50 beats/min and <100 beats/min; and 6) have a Glasgow Coma Scale score of 15. CONCLUSIONS: This prediction rule for safe early discharge of patients with presumed opioid overdose performs well in this derivation set but requires validation followed by confirmation of safe implementation. PMID- 11015243 TI - Maximizing the sensitivity and specificity of pediatric trauma team activation criteria. AB - BACKGROUND: Care of the severely injured child requires the rapid assembly of personnel trained in pediatric trauma care. Trauma team activation criteria, which are highly sensitive and maximally specific for identifying the child who requires resuscitation, are necessary to provide rapid care to all who need it, while using resources efficiently. OBJECTIVE: To determine the sensitivity and specificity of the standard trauma team activation (TTA) criteria for identifying patients who receive resuscitation in the emergency department. METHODS: A one year study was conducted of all patients transported by emergency medical out-of hospital services for a trauma-related complaint. For all patients, out-of hospital medical control operators recorded whether patients met TTA criteria and, if so, which criteria were met. Criteria included standard physiologic, anatomic, and mechanism parameters. Sensitivity and specificity for the outcome of resuscitation (volume restoration, assisted ventilation or intubation, chest tube insertion/needle decompression, operative intervention) were calculated. RESULTS: A total of 492 patients met the case definition. Two-thirds were male, the mean age was 8 years (+/-4.8 SD), and the Injury Severity Score was > or =15 in 9.3%. Trauma team activation criteria were met by 179 patients (36. 4%) and, of these, 107 met mechanism criteria only. A resuscitative intervention was received by 54 (10.9%) of the total and none in the mechanism-only group. Sensitivity and specificity of the TTA criteria for predicting receipt of a resuscitation procedure were 98. 1% and 71.2%, respectively. When mechanism criteria were excluded, the sensitivity remained 98.1% and the specificity increased to 95. 7%. CONCLUSIONS: Criteria for TTA that include patients who meet mechanism criteria only are not specific for identifying patients who receive a resuscitative intervention. Use of anatomic and physiologic criteria only results in an increase in specificity, thereby reducing overtriage while retaining a high sensitivity. PMID- 11015244 TI - Conjoint smoking and drinking: a case for dual-substance intervention among young emergency department patients. AB - OBJECTIVES: To better understand conjoint smoking and drinking among young adult emergency department (ED) patients, the purposes of this investigation were: 1) to assess the prevalence of conjoint use; 2) to determine the factors associated with conjoint alcohol use and smoking; and 3) to address the implications for future ED-based investigation of dual-substance intervention. METHODS: Data for this investigation were obtained from a battery of questionnaires administered to the routine-care patients during an alcohol screening in the ED, which was part of a larger alcohol intervention study. RESULTS: Study findings revealed that a majority of patients with self-reported alcohol-related problems were smokers. In fact, drinkers who smoked were likely to be pack-a-day smokers. Among the study sample, being female, having low education levels (e.g., high school education or less), having some emotional problems, and currently using marijuana were risk factors for conjoint smoking and drinking. CONCLUSIONS: Conjoint users were identifiable through brief screening. Given the prevalence of conjoint smoking and alcohol use among the ED sample and a specific set of risk factors, tailored intervention for alcohol and nicotine dependence may be an important and opportunistic clinical ED service. PMID- 11015245 TI - Emergency medicine resident documentation: results of the 1999 american board of emergency medicine in-training examination survey. AB - OBJECTIVES: To assess how emergency medicine (EM) residents perform medical record documentation, and how well they comply with Health Care Financing Administration (HCFA) Medicare charting guidelines. In addition, the study investigated their abilities and confidence with billing and coding of patient care visits and procedures performed in the emergency department (ED). Finally, the study assessed their exposure to both online faculty instruction and formal didactic experience with this component of their curriculum. METHODS: A survey was conducted consisting of closed-ended questions investigating medical record documentation in the ED. The survey was distributed to all EM residents, EM internal medicine, and EM-pediatrics residents taking the 1999 American Board of Emergency Medicine (ABEM) In-Training examination. Five EM residents and the Society for Academic Emergency Medicine (SAEM) board of directors prevalidated the survey. Summary statistics were calculated and resident levels were compared for each question using either chi-square or Fisher's exact test. Alpha was 0.05 for all comparisons. RESULTS: Completed surveys were returned from 88.5% of the respondents. A small minority of the residents code their own charts (6%). Patient encounters are most frequently documented on free-form handwritten charts (38%), and a total of 76% of the respondents reported using handwritten forms as a portion of the patient's final chart. Twenty-nine percent reported delays of more than 30 minutes to access medical record information for a patient evaluated in their ED within the previous 72 hours. Twenty-five percent "never" record their supervising faculty's involvement in patient care, and another 25% record that information "1-25%" of the time. Seventy-nine percent are "never" or "rarely" requested by their faculty to clarify or add to medical records for billing purposes. Only 4% of the EM residents were "extremely confident" in their ability to perform billing and coding, and more than 80% reported not knowing the physician charges for services or procedures performed in the ED. CONCLUSIONS: The handwritten chart is the most widely used method of patient care documentation, either entirely or as a component of a templated chart. Most EM residents do not document their faculty's participation in the care of patients. This could lead to overestimation of faculty noncompliance with HCFA billing guidelines. Emergency medicine residents are not confident in their knowledge of medical record documentation and coding procedures, nor of charges for services rendered in the ED. PMID- 11015246 TI - Clinicopathological conference: sudden-onset facial edema in an autistic child. PMID- 11015247 TI - Organization of emergency medicine at medical schools: compelling reasons for departmental status. PMID- 11015248 TI - Ethics seminars: case studies in "futility"-challenges for academic emergency medicine. AB - The concept of "futility" and its determination in emergency medicine pose unique challenges to emergency physicians, patients, and society. The term "futility," although commonly used, is problematic in its scope, meaning, and interpretation. To bridge this gap in understanding, the authors suggest the construct of clinically nonbeneficial interventions (CNBI), instead of "futility. " This language better informs discussions of nonbeneficial interventions across the risk spectrum of emergency medical practice, while retaining the focus on the patient's interests. Two cases are presented, which underscore the need for prudence and empathetic communication when addressing issues of CNBI. Determinations of expected benefit should be based on established scientific evidence, and the goals and values of patients, not on individual biases regarding quality of life or other subjective matters. While physicians are under no ethical obligation to provide treatments that they judge have no realistic likelihood of clinical benefit, the context in which these determinations take place is of critical importance. When certain interventions are appropriately withheld, concerted efforts should be made to maintain effective communication, comfort, support, and counseling for patients, friends, and families. In all aspects of clinical decision making, the value of various interventions and therapies must be based on expected risks and benefits to the patients, first and foremost. PMID- 11015249 TI - Can clinical parameters predict fractures in acute pediatric wrist injuries? AB - OBJECTIVE: Fractures around the wrist are common in pediatric patients presenting to the emergency department (ED). This pilot study was aimed at identifying clinical variables that are most likely to be associated with a fracture. METHODS: This was a prospective blinded case series of patients 3-18 years of age presenting with an acute (<3 days) wrist injury, without obvious deformity. A team of five investigators blinded to the eventual radiographic findings evaluated patients. Physical examination variables included range of motion (ROM), site of maximal tenderness, and functional deficit. The latter was determined objectively, by recording any difference in grip strength between the injured and noninjured hands. Diagnostic radiographs were obtained for all patients. Univariate analysis using Wilks' log likelihood ratio test was performed to identify clinical variables associated with confirmed wrist fractures. Sample size was determined based on the ability to detect a difference of 15 degrees in the ROM variables, 20% point differences in grip strength, and 30% proportion differences in categorical variables using a power of 0.8 and a two-tailed of 0.05. RESULTS: The ROMs were not significantly different between the fracture (Fx) and nonfracture (NFx) group. There was significant change in the grip strength between the Fx and NFx groups (t = 3.3, p = 0.0019). Tenderness over the distal radius was also associated with a greater likelihood of a fracture (G(2) = 5.0, p = 0.02). Sensitivity of clinical prediction was found to be 79%, and specificity was 63%. The false-negative rate was 0.21 and the false positive rate was 0.37, while the positive predictive value was found to be 0.68 and negative predictive value 0.75. CONCLUSIONS: Distal radius point tenderness and a 20% or more decrease in grip strength were predictive of fractures. PMID- 11015250 TI - Low plasma thiamine levels in elder patients admitted through the emergency department. AB - OBJECTIVES: To determine the prevalence of thiamine deficiency in a high-risk group of elder emergency department (ED) patients who reside in nursing homes and need admission to the hospital, and to determine the effect of patients' diets on this prevalence. METHODS: This was an observational pilot study of 75 consecutive ED patients aged 65 years or older who lived in a nursing home and were admitted to the hospital. Plasma thiamine levels were measured by high-pressure liquid chromatography on serum samples collected within 24 hours of hospital admission. Nursing home records were reviewed to determine whether patients received nutritional supplementation or enteral tube feedings. RESULTS: Seventy patients participated and had a mean plasma thiamine level of 27.3 microg/dL (95% CI = 20.2 to 34.4). Fourteen percent (n = 10, 95% CI = 8% to 24%) were thiamine deficient (<10 microg/dL). Patients not receiving dietary supplements or tube feedings (n = 26) had lower mean thiamine levels (20.3 microg/dL, 95% CI = 12.7 to 27.9) and were thiamine-deficient more often (27%) than patients receiving dietary support (n = 44, 31.5 microg/dL, 95% CI = 24.7 to 38.3, 7% thiamine deficient). CONCLUSIONS: Elder nursing home patients seen in the ED and admitted to the hospital are frequently thiamine-deficient. Empiric thiamine supplementation is often used in the ED for other high-risk patients, such as alcoholic individuals, and may be appropriate for high-risk elder patients. Further research is needed to determine whether thiamine supplementation in these patients can improve their clinical outcomes. PMID- 11015251 TI - Bilateral tubal ectopic pregnancy: a tale of caution. AB - Diagnosis of ectopic pregnancy continues to be an important challenge facing emergency physicians. The authors present a case of bilateral tubal ectopic pregnancy and discuss its clinical features and diagnostic difficulties. A review of the English-language literature on the subject is discussed. Suggestions are made on ways to increase diagnostic accuracy, reduce complications, and preserve future fertility in this group of patients. PMID- 11015252 TI - The accuracy of the emergency physician at diagnosing CVA/TIA in the acute care setting AB - OBJECTIVE: To determine the accuracy of the ED physician at diagnosing CVA/TIA in the acute setting. METHODS: We reviewed 246 patients admitted for acute CVA/TIA during 1997. We reviewed admitting and discharge diagnoses, CT, MRI, and MRA results. We also reviewed the medical histories of the patients. Patients with tumors and ICH diagnosed on the initial CT scan were excluded. RESULTS: A total of 241 patients were included for analysis. Of the patients admitted for CVA, 67% were discharged with the same diagnosis. Of the patients admitted for TIA, 82% were discharged with the same diagnosis. 10% of TIAs diagnosed at admission received the diagnoses at discharge. 22% of CVAs at admission were diagnosed as TIAs. 11% of CVA/TIAs at admission were given other diagnoses at discharge. Some of these diagnoses included hemiplegic migraines, Bell's palsy, lumbosacral spondylosis, giant cell arteritis, basilar artery aneurysm, and viral meningitis. In considering thrombolysis for CVA, one may then overtreat approximately 30% of patients and miss approximately 10% of patients who could be candidates for treatment. The NNT for CVA and thrombolysis is approximately 10. If 30% overtreatment rate is accurate, then the NNH is approximately 333 assuming an ICH rate of 1%. For 1,000 patients treated, 100 may benefit (improved function) and 3 would die who did not have the disease. CONCLUSIONS: The accuracy rate of the diagnosis of CVA at initial presentation is 67%. This is due to many initial neurologic changes being TIAs and some other diagnoses which can mask as CVA. Choosing thrombolysis for CVA treatment will involve treatment of many patients who do not have disease. PMID- 11015253 TI - EMS transports for difficulty breathing: is there a potential role for CPAP in the prehospital setting? AB - Mask-applied continuous positive airway pressure (CPAP) has been shown to reduce morbidity among patients with acute respiratory distress in the setting of cardiogenic pulmonary edema. OBJECTIVE: To determine a minimum percentage of patients transported by ALS for difficulty breathing who could potentially benefit from a pre-hospital trial of CPAP. METHODS: Paramedic run sheets were collected from consecutive, adult, ALS transports for a chief complaint of difficulty breathing over a 6 week period in a large urban EMS system. Demographic information, medical history, vital signs, clinical assessments, and transport times were abstracted into a database by trained reviewers. Strict criteria for CPAP were defined in advance as "acute respiratory distress," meaning (1) respiratory rate > 25 and (2) labored or shallow breathing, and "presumed cardiogenic pulmonary edema," meaning (3) a prior history of heart disease and (4) presence of bilateral rales on exam. RESULTS: Data from 240 consecutive run sheets were compiled. Median patient age was 66 years old, with females outnumbering males 168 to 81. A total of 15 spontaneously breathing patients met all 4 criteria for CPAP. Four of these patients were either hypotensive (SBP < 90) or had potential for airway compromise (i.e., obtundation), making CPAP inadvisable. Among the 11 remaining patients (4.4% of all transports for difficult breathing), median transport time was 20 minutes (range 14-31 minutes). CONCLUSIONS: Using very strict criteria, a small but not significant percentage of patients are optimal candidates for a prehospital trial of CPAP. Transport times would appear to justify this type of intervention. A prospective study is currently under way to test the feasibility of administering CPAP to such patients in the prehospital setting. PMID- 11015254 TI - Etomidate-facilitated hip reduction in the emergency department AB - OBJECTIVE: The authors report what they believe is the first reported use of etomidate to assist in the reduction of a major joint in an outpatient setting. METHODS: The authors review the case of an elderly woman with a total hip arthroplasty who experienced four spontaneous posterior hip dislocations in a 5 month period. Etomidate was successfully used in two dislocations where previous methods had failed. RESULTS: A 68-year-old woman, who 13 months earlier had an uncomplicated total left hip replacement, was transported to the same ED on four separate occasions for a spontaneous left hip dislocation. Radiographs in each instance were significant only for a posterior dislocation of the implant articulation. The first reduction in the ED was unsuccessful and required a closed reduction in the operating theater. Seventy-two hours later the second dislocation and subsequent reduction occurred in the ED using etomidate to facilitate muscle relaxation. The patient was subsequently discharged home. Similar scenarios were replayed in the next few months. CONCLUSIONS: Muscle relaxation is the key characteristic for the reduction of dislocated major joints. The risks of respiratory depression and hemodynamic alterations with sedation are not insignificant, especially at the extremes of age. In the present case, intravenous narcotics and sedative-amnestic agents did not result in sufficient muscle relaxation. Larger or repeated doses may have resulted in undesirable or dangerous side effects. Etomidate is a useful adjunct when cardiopulmonary disease is present. The rapid onset and recovery from etomidate make it an excellent choice for facilitating the reduction of hip dislocations in elderly patients with prior total hip replacements. PMID- 11015255 TI - Antioxidant treatment improves bioenergetics following Ischemia/Reperfusion AB - Published in: Acad Emerg Med. 2000; 7: 507-8. PMID- 11015257 TI - Accuracy of interqual criteria in determining the hospitalization need in medicare patients with gastrointestinal bleeding AB - Published in: Acad Emerg Med. 2000; 7: 552-3. PMID- 11015256 TI - Trauma in the very elderly: A community-based study of outcomes at trauma and non trauma centers AB - Published in: Acad Emerg Med. 2000; 7: 568. PMID- 11015258 TI - Retrospective evaluation of potential medicare admission denials using interqual and millman and roberts admission criteria AB - Published in: Acad Emerg Med. 2000; 7: 543. PMID- 11015259 TI - Prevalence and documentation of impaired mental status in elderly emergency department patients AB - OBJECTIVE: To determine the prevalence and assess documentation by emergency department (ED) physicians (EPs) of impaired mental status in elderly ED patients. METHODS: Cross-sectional, observational study. Subjects: convenience sampling of ED patients greater than or equal to 70 years of age. Patients were screened for cognitive impairment with the Orientation Memory Concentration exam (OMC), and for delirium with the Confusion. Assessment Method screening tool (CAM). A positive OMC or CAM was considered indicative of impaired mental status. Patients with delirium were excluded from the cognitive impairment screen. EPs were blinded to screening results. Physician documentation, dispositions, and referrals were abstracted from chart review. Proportions and 95% confidence intervals (CIs) are reported. RESULTS: 180 patients were screened. 46 patients (26%; 95% CI = 19% to 32%) had impaired mental status. 22 of these (12%; 95% CI = 7% to 17%) had delirium, and 24 (13%; 95% CI = 8% to 18%) had moderate to severe cognitive impairment. Of all patients with impaired mental status, only 14 (30%; 95% CI = 18% to 46%) had documentation of any impairment by the EP (10 with delirium (46%; 95% CI = 24% to 68%), and 5 with cognitive impairment (21%; 95% CI = 7% to 42%). 7 of 22 (32%; 95% CI = 14% to 55%) patients with delirium were discharged home. Only 2 of 16 patients with impaired mental status (12.5%) were discharged home with plans noted to address the impairment. CONCLUSIONS: Impairment in mental status is highly prevalent among older ED patients. Lack of documentation and referrals by EPs suggests lack of recognition of these problems. Further education of physicians is needed to improve care in these areas. PMID- 11015260 TI - Case finding of At-risk elders in the emergency department (ED): A multicenter study AB - Published in: Acad Emerg Med. 2000; 7: 563-4. PMID- 11015261 TI - Emergency medicine residencies: A descriptive study of program structure AB - The steady growth in EM residency programs reflects the desirability of EM as a specialty area for training. However, a profile of programs offered nationwide is not available. OBJECTIVE: To characterize emergency medicine (EM) programs nationwide according to program format, curriculum, and opportunities for subspecialty training. METHODS: In this descriptive study, U.S. emergency medicine residency curriculums were evaluated electronically using SAEM and FRIEDA residency information websites. Data collected included program format, hospital size and volume, rotation durations, curriculum and electives offered. RESULTS: The most common format (72%, n = 87) is PGY 1, 2, 3 and PGY 2, 3, 4 accounted for 18% (n = 22) and PGY 1, 2, 3, 4 for 10% (n = 12). The average length of training in the ED is 19 months (range 7-40). Average general pediatric training is 1.8 months (range 0-5). The average number of rotations outside the primary site is 4.9 (range 0-19) and elective rotation time and topics varied considerably. Opportunities for air medical training differed by program format (82% of PGY 1-3, 79% of PGY 1-4, 75% of PGY 2-4) (table 11-1). CONCLUSIONS: There is substantial variation in curriculum, in subspecialty exposure, and in opportunity for electives for residents depending on program format and location. PMID- 11015262 TI - A critical examination of the USMLE II: does a study month improve test performance? AB - Students often are encouraged to elect a study month prior to taking the USMLE II. OBJECTIVE: To determine if a study month prior to the September USMLE II improves student test performance. METHODS: Student clerkship sequence was prospectively recorded for each senior at a large midwestern medical school during the 1998-99 academic year, and USMLE I and II scores were obtained. USMLE II scores were compared between students who elected a study month prior to the September testing date versus those who elected clerkships, while controlling for USMLE I test scores using multiple linear regression. RESULTS: Complete data were obtained for 167 senior year medical students. 124 (74.3%) students took the September USMLE II after electing a study month, and 43 (25.7%) students took the September USMLE II after electing clerkships. The mean score for those who elected a study month before the USMLE II test was higher than those electing clerkships (210.56 +/- 17.12 vs 208.65 +/- 20.83). However, after controlling for USMLE I scores (r(2) = 0.53; p < 0.01), the election of a study month prior to the September USMLE II was not associated with an increase in student performance (r(2) < 0.01; p = 0.73). CONCLUSIONS: These results do not support study months as a means to improve USMLE II test performance, and suggest that rotations may be equally beneficial. PMID- 11015263 TI - Inhaled furosemide in the treatment of acute exacerbations of asthma AB - Inhalation of the loop diuretic furosemide has been found to protect against challenges with bronchoconstrictive agents. OBJECTIVE: To determine if furosemide has any objective and/or subjective therapeutic effect on adult patients with acute exacerbations of asthma, above and beyond the effects of B2 agonists and oral glucocorticoids. METHODS: 35 adult patients with acute exacerbations of asthma were randomized and recruited into the study if they had a prior history of asthma, and <10 pack-year smoking history, had not received pre-hospital B2 agonists, were not pregnant, and had no sulfonamide allergy. A double blind study design was employed. One group received albuterol 5 mg with placebo and one group received albuterol 5 mg with furosemide 40 mg in their nebulizer. Both groups received prednisone 80 mg po. PEFRs were obtained before initiation of treatment and 30 and 60 minutes thereafter. A 10 point dyspnea scale was also employed at initiation of treatment and 30 and 60 minutes thereafter. Physicians were permitted to give rescue albuterol treatments as deemed necessary. RESULTS: There was no difference in the treatment arms with respect to PEFR, dyspnea score, or number of rescue albuterol treatments at 30 and 60 minutes. CONCLUSIONS: We found no statistical benefit to adding furosemide to a regimen consisting of prednisone and optimal doses of B2 agonist. We conclude that there is not a current role for furosemide in acute exacerbations of asthma. PMID- 11015264 TI - Changes in stroke index measured by impedance cardiography in a human model of moderate acute blood loss AB - Early detection of acute hemorrhage is problematic because changes in traditional parameters such as heart rate and blood pressure are notoriously unreliable. OBJECTIVE: To observe the effect of acute, moderate blood loss on stroke index (SI) as measured non-invasively by impedance cardiography. METHODS: Five healthy, adult volunteers were phlebotomized 590 mL over ten minutes. The volume of blood loss was expressed as a percent of estimated total blood volume (TBV) according to the formula TBV = (weight in kg) x (70 mL/kg). Stroke index was measured in a fixed, reclining position before (SI #1) and after (SI #2) phlebotomy, using the BioZ.com (Cardiodynamics International Corporation, San Diego, CA) thoracic bioimpedance monitor. Absolute and relative changes were calculated. RESULTS: In each volunteer, a fall in SI was noted after phlebotomy. The percent change in SI ranged from 8% to 18% (table 15-1). There appeared to be a correlation between blood loss as a percent of estimated total blood volume and the percent decline in SI among these volunteers. CONCLUSIONS: Impedance cardiography reliably detected moderate acute blood loss in this small set of healthy, adult volunteers. If these results can be quantified and validated prospectively in a larger population, SI may prove to be a valuable, non-invasive parameter for detection of early hemorrhage shock. PMID- 11015265 TI - Compliance with emergency department prior authorization requirements under EMTALA AB - Published in: Ann Emerg Med. 1999; 34(4 pt 2):S70. PMID- 11015266 TI - The role of taurine for cellular volume regulation of the hippocampus AB - Taurine is used for volume regulation in a number of cultured cells, including neurons and glia. OBJECTIVE: To evaluate the role of this amino acid in brain volume regulation in situ. METHODS: Hippocampi of adult Sprague-Dawley rats were cut into 400 &mgr;m slices and equilibrated for 2 hr at room temperature in artificial cerebrospinal fluid (aCSF) bubbled with 95% O(2)/5% CO(2). Slices then were perfused with aCSF at 36 degrees C and optically imaged. After 1 hr in isotonic aCSF (290 mOsm), slices were perfused with hypoosmotic aCSF (200 mOsm) made by reducing the concentration of NaCl. Images of transmitted light intensity were captured every 60 sec to assess intracellular volume. For some slices, 1 mM taurine was added to aCSF during room temperature equilibration and during the first 30 min at 36 degrees C to replace that lost during slice preparation. Taurine contents were measured in perchlorate extracts by HPLC. Data were analyzed by Student's t-tests. RESULTS: Taurine contents of slices incubated in isotonic aCSF with and without taurine were 97 +/- 1 and 26 +/- 2 nmol/(mg protein), respectively. The intensity of transmitted light through the CA1 region increased similarly during the first 20 min in hypoosmotic aCSF, regardless of taurine treatment. However, after 60 min of hypoosmotic exposure, light transmittance recovered more in taurine-treated slices (44 +/- 6%) than in untreated slices (28 +/- 3%, p <0.05). The hypoosmotic-induced transmittance increase was smaller in the CA3 region for both groups of slices; however, similar effects of taurine treatment were obtained for subsequent transmittance recovery. CONCLUSIONS: Sufficient levels of taurine are required for effective volume regulation of hippocampal slices. PMID- 11015267 TI - Comparison of two intraosseous infusion techniques in an EMT training program AB - OBJECTIVE: To compare placement of IO lines using a traditional method with placement using a new device called the bone injection gun (BIG). METHODS: A prospective cross-over study was conducted to compare the time to line placement and ease of insertion for both traditional (Jamshidi) and BIG methods. EMT-P students and practicing paramedics provided information about their previous experience with IO line placement. They were assigned to establish an IO line in a pediatric leg mannequin using each technique, and rated the ease-of-use of each method. RESULTS: Thirty-eight participants (28 EMT-P students and 10 paramedics) completed both of the IO skills. Only two (5%) had ever placed an IO line in a patient previously. Time to placement of the IO device was faster in the BIG group (16.91 sec. vs. 11.93 sec., p = 0.02). There was no statistical difference in ease-of-use ratings between the methods (p = 0.816). Student times for establishing the IO line with the BIG device were faster than the times of the practicing paramedics (11. 18 sec. vs. 16.25 sec., p = 0.049). CONCLUSIONS: In the education setting, times to establishment of an IO line were similar using both the traditional method of insertion and the BIG device. Ratings of both the students and paramedics were similar with respect to each-of-use for both the methods. PMID- 11015268 TI - Weight of backpacks carried by elementary school children: students or sherpas? AB - Published in: Acad Emerg Med. 2000; 7: 487. PMID- 11015269 TI - An assessment of quality control testing in an emergency department (ED) maintained arterial blood gas analyzer AB - Emergency medicine by its nature requires the immediacy of laboratory testing. Point of care testing (POCT) affords the ED the fastest access to important laboratory data. To the best of the authors' knowledge, maintenance of an arterial blood gas (ABG) analyzer as a POCT device by ED personnel has not been studied. OBJECTIVE: To determine the accuracy of POCT quality control testing of an ED maintained ABG Analyzer (Gem Premier)-Instrumentation Laboratory). METHODS: We compared comprehensive quality control data obtained when the ABG analyzer was maintained by the Department of Clinical Laboratories with data obtained from the same analyzer when it was maintained by ED attending physicians. Each parameter measured by the ABG analyzer was tested using unknown control samples supplied by the College of American Pathologists. These two groups of data were then compared to the total number of obtained values submitted to the College of American Pathologists. RESULTS: Each parameter measured (n = 7) showed no difference between the two groups in regard to overall pass rate. All values for the two groups fell within the limits of acceptability when compared to all other samples analyzed by the College of American Pathologists (range 159-208 other labs). CONCLUSIONS: ED attendings are capable of quality control maintenance of an ABG analyzer as a POCT device under the quality control testing parameters set forth by the College of American Pathologists. PMID- 11015270 TI - Diagnostic odor recognition AB - Many diseases, toxic ingestions, and intoxications have characteristic odors. These odors may provide diagnostic clues that affect rapid treatment long before laboratory confirmation or clinical deterioration. Odor recognition skills, similar to auscultation and palpation skills, require teaching and practical exposure. Dr. Goldfrank and colleagues recognized the importance of teaching odor recognition to emergency service providers. They proposed the "sniffing bar" method for odor recognition training. OBJECTIVES: (1) To identify the recognition rates of medically important odors among emergency care providers. (2) To investigate the effectiveness of teaching odor recognition. Hypothesis: The recognition rates of medically important odors will increase after teaching exposure. METHODS: The study exposed emergency care providers to 11 tubes of odors. Identifications of each substance were recorded. After corrective feedback, subjects were re-tested on their ability to identify the odors. Analysis of odor recognition improvement after teaching was done via chi-square test. RESULTS: Improvement in identification after teaching was seen in all odors. However, the improvement was significant only in the lesscommon substances because their initial recognition was especially low. Significant changes may improve with a larger sample size. Subjects often confuse the odors of alcohol with acetone, and wintergreen with camphor. CONCLUSIONS: The recognition rates are higher for the more-common odors, and lower for the less-common odors. Teaching exposures to the less well-known odors are effective and can significantly improve the recognition rate of these substances. Because odor recognition may affect rapid diagnosis and treatment of certain medical emergencies such as toxic ingestion, future studies should investigate the correlation between odor recognition and the ability to identify corresponding medical emergencies. PMID- 11015271 TI - Physiological manifestations of systemic activation of complement AB - Published in: Acad Emerg Med. 2000; 7: 558. PMID- 11015272 TI - Shaken. Not stirred-temperature change and heat loss during delivery of IV fluids AB - Microwave-heated intravenous fluids are used in the rewarming of hypothermic patients. OBJECTIVES: To study the effect of both shaking of the microwaved bag and heat loss during delivery. METHODS: Twenty 1-liter normal saline bags were heated individually in a commercial microwave, immediately randomized into a "shaken" or a "non-shaken" group. The temperature of the fluid was recorded initially out of the bag and then at one-minute intervals by a blinded observer as the fluid ran "wideopen" through ambient temperature tubing. RESULTS: No statistically significant temperature difference occurred in any of the measured time intervals between the shaken and the non-shaken bags. Seventy percent of the overall temperature losses occurred in the first three minutes out of the microwave for both groups. CONCLUSIONS: Absence of shaking of the microwaved fluids does not produce "hot spots." Higher initial temperatures out of the bag should be considered as well as warming of the IV tubing. PMID- 11015273 TI - The effect of carbonated beverages on colorimetric end-tidal CO(2) determination AB - Esophageal intubation is a significant complication of attempted airway control. Colorimetric end-tidal CO(2) monitors are highly sensitive and specific for detecting the presence of CO(2). There are reports of false-positive end-tidal CO(2) readings from esophageal intubations in patients who had ingested carbonated beverages. OBJECTIVE: To determine whether carbonated gastric contents can affect colorimetric end-tidal CO(2) readings. METHODS: End-tidal CO(2) was measured in sacrificed piglets at measured time intervals after instillation of 5 , 10-, 15-, and 20-mL aliquots of Diet Coke were placed into the stomach via a 28 French orogastric tube followed by esophageal intubation. The stomach was completely evacuated prior to each instillation and rechecked for baseline CO(2). RESULTS: Compiled data from three piglets (20-30 kg). Piglets were not ventilated or moved and determinations were measured at 20 degrees while supine. All data collected within 2 hours postmortem (table 24-1). CONCLUSIONS: The colorimetric end-tidal CO(2) turned "yellow" and did not change to blue with extended insufflations. The CO(2) of a small quantity of carbonated beverage in the stomach could be "blown off" by multiple insufflations. We conclude that esophageal intubation in the setting of recent ingestion of a carbonated beverage may result in a false-positive end-tidal CO(2) determination. PMID- 11015275 TI - Efficacy of the porcine model for demonstrating pericardial effusion with real time ultrasound AB - Ultrasound in emergency medicine (EM) is an invaluable tool for emergent evaluation of pericardial effusion and tamponade. The training in the recognition of such has relied on ultrasonic images or videotapes. OBJECTIVE: To evaluate the efficacy of demonstrating pericardial effusion and tamponade using real-time ultrasound. METHODS: A convenience sample of 10 Yorkshire swine were anesthetized, sedated, intubated, and monitored as per an approved EM training protocol. Using EKG guidance and a subxiphoid approach, an 18-gauge spinal needle was advanced into the pericardium until a current of injury was detected. After a slight withdrawal of the needle, an aliquot of 50-100 mL of normal saline was instilled to produce a pericardial effusion. Ultrasound was used before and after the instillation of fluid into the pericardium. The effusion was confirmed by subsequent pericardiocentesis or by visual inspection on thoracotomy. RESULTS: The instillation saline, detection of effusion, and subsequent confirmation by ultrasound, pericardiocentesis, or visual inspection were successfully performed in 10/10 pigs. Under ultrasound, one pig had a preexisting effusion. Another had much less fluid visualized. On thoracotomy, the saline was found to have leaked into the adjacent lung parenchyma. CONCLUSIONS: We conclude that the porcine model is an adequate model to demonstrate pericardial effusion using real-time ultrasound. PMID- 11015274 TI - Ultrasonography evaluation of retrobulbar hematoma in bovine orbits AB - As the proptotic eye is pushed anteriorly against the canthis, pressure within the eye increases until there is inadequate perfusion of the eye due to the pressure effects. computed tomography (CT) scan of the orbit is the most common method of diagnosing a retrobulbar hematoma. In the trauma/emergency department setting, where the wait for a CT scan may present significant delay, bedside ultrasonography may aid in the rapid diagnosis and treatment of a retrobulbar hematoma. OBJECTIVE: To establish concordance between orbital CT scan and a bedside sonographic orbital evaluation of a retrobulbar hematoma. METHODS: Eighteen of 22 bovine orbits were injected with a 10-mL mixture of calcium treated banked blood and contrast into the retro-orbital space thereby effecting a retrobulbar hematoma. Each orbit was studied with CT and evaluated by a neuroradiologist. Four orbits were left intact as controls. Each orbit underwent an ultrasonographic evaluation. The ultrasonographer was first allowed to study an orbit with a documented hematoma and a control. Thereafter, he was blinded to the orbits containing hematoma, did not physically examine each orbit, and completed each exam in a darkened room. The CT results were correlated with the ultrasound findings. RESULTS: 15 of 18 orbits (83%) were correctly identified as containing hematoma by ultrasound. All four intact orbits were correctly identified as being free of hematoma (kappa 0.6). CONCLUSIONS: Bedside ultrasonography may aid in the rapid diagnosis of a retrobulbar hematoma. PMID- 11015276 TI - Blood cultures in pyelonephritis: Do results change therapy? AB - OBJECTIVE: To determine how often blood culture results are instrumental in directing the therapy of patients with pyelonephritis. METHODS: Retrospective chart review of all patients admitted to Doctors Hospital with a diagnosis of pyelonephritis to ascertain whether the results of blood cultures were instrumental in the management of those patients. For study purposes, results of blood cultures were considered instrumental if the treatment regimen or diagnostic procedures were changed because of those results. RESULTS: 212 patients were admitted during the study period with a diagnosis of pyelonephritis. Of these, 105 (49.5%) had blood cultures ordered as part of their workup. Adult patients had at least 2 blood cultures ordered and some had as many as 4 or 6 sets ordered during hospitalization. Most pediatric patients had only a single blood culture ordered for diagnosis. Of the 105 patients, 88 (83.5%) had no growth on any of the tubes drawn. 17 patients had positive growth in at least one tube; however, 6 out of these 17 patients were judged by the treating physician to have contaminated cultures that were not diagnostically useful. Of the 105 patients, only 11 (10.4%) were considered to have positive culture results by their treating physician. The organism identified by blood culture was compared to that present in the patient's urine culture, and in only 2 patients did the results differ and were found to have a secondary site of infection. Only in these 2 cases were the antibiotic regimens changed because of the blood culture results. No patient without a second site of infection had a change in therapy based on the blood culture results. CONCLUSIONS: In patients admitted for pyelonephritis, blood cultures may not be needed. PMID- 11015277 TI - The incidence of positive cultures in women suspected of having PID/Salpingitis AB - OBJECTIVES: To determine the incidence of GC and chlamydia in women with suspected acute pelvic inflammatory disease. A secondary objective was to investigate the clinical usefulness of physical exam findings of vaginal discharge, cervical motion, and adnexal tenderness. METHODS: This was a retrospective chart review of patients seen in the ED. Patients were entered into the study if they had PID/salpingitis as their discharge diagnosis and cultures performed for GC and chlamydia. RESULTS: A total of 133 charts were reviewed. 13 patients were excluded due to incomplete charting of history and physical exam or inability to obtain culture results. Of the remaining 120 patients, 70 cultures were negative for any growth. 10 cultures were positive for GC and 10 cultures were positive for chlamydia. In reviewing the physical exam findings in women with negative cultures, 74% had discharge, 93% had cmt, and 66% had adnexal tenderness. For women with cultures positive for GC and chlamydia, 90% had discharge, 80% had cmt, and 75% had adnexal tenderness. For vaginal discharge the sens, spec, ppv, npv were 0.90, 0.26, 0.35, 0.90. For cmt the sens, spec, ppv, npv were 0.80, 0.07, 0.20, 0.55. For adnexal tenderness the sens, spec, ppv, npv were 0. 75, 0.34, 0.25, 0.83. No combination of the three physical exam findings increased the ppv. CONCLUSIONS: The incidence of positive cultures in women suspected of having PID/salpingitis is 16%. The physical exam findings were not predictive of disease secondary to the high number of false positives. Although 16% positives may warrant empiric treatment, patients should not be told they have PID/salpingitis until the cultures are available. PMID- 11015278 TI - Emergency medicine residency director perceptions of the resident selection process AB - OBJECTIVE: To assess perceptions of emergency medicine (EM) program directors (PDs) of the resident selection process. METHODS: PDs of 120 EM residencies were mailed a 22 question survey immediately following the 1999 match. (Applicants interviewing in more than one specialty and post-interview communications were areas of interest). Non-respondents received a second mailing. RESULTS: Ninety eight (81. 7%) completed surveys were returned. Five PDs felt that interviewing in more than one specialty was ethically wrong, while most felt multi-specialty interviews were a system byproduct (42%) or that there was nothing wrong (41%). Eighty-one percent of PDs felt the applicant's rank order would be negatively affected if the applicant disclosed interviewing in >1 specialty. Forty-seven percent of PDs (always or frequently) told applicants to keep in touch if interested in the program. However, 88% of PDs were skeptical or did not believe an applicant's communicated intent to rank the program 'high,' nor did this communication influence an applicant's rank order (75%). Forty-two percent of PDs reported informal commitments by applicants. PDs frequently felt lied to by applicants (always (4%), frequently (42%), sometimes (42%)). Applicants often ask how the program intends to rank them. Highly ranked applicants receive positive responses from 61% of PDs vs. 33% of PDs who give negative responses to low ranked applicants. Ten percent of PDs offer residency positions outside of the match. CONCLUSIONS: Applicants who interview in >1 specialty are viewed negatively by PDs. Post-interview communications by applicants are viewed with skepticism. Gamesmanship is practiced commonly during the resident selection process. PMID- 11015279 TI - Medical toxicology experience during emergency medicine residency AB - Medical toxicology (MT) is a core content area of emergency medicine (EM) as well as an American Board of Medical Specialties recognized subspecialty of EM. OBJECTIVE: To obtain information concerning MT education in EM residencies. METHODS: A questionnaire was sent to the director of each EM program in the Society for Academic Emergency Medicine (SAEM) Residency Catalog. Subsequent telephone contact was made to encourage participation. Information was obtained about program format, poison control center (PCC) location, and inclusion of a toxicologist on faculty as well as specifics about MT rotations and organized educational activities. RESULTS: Survey participation was 62 of 122 (51%) programs. A toxicologist is on faculty at 39 (63%) programs and 5 (26%) of the programs without a toxicologist intend to recruit one in the near future. A MT rotation is a requirement of 46 (76%), an elective at 12 (19%), and not available at 3 (5%) programs. The rotation is >/=4 weeks at 46 (78%) of the programs with a rotation. At 18 (31%), time is spent primarily at the hospital, while at 26 (44%) it is at the PCC. At 5 (8%) programs, equal/near equal time is spent at the hospital and PCC. The individual(s) providing resident supervision at PCCs is not uniform. Organized MT experience other than a rotation is included in 39 (63%) of programs. CONCLUSIONS: Due to suboptimal participation, many issues regarding MT education in EM residency remain uncertain. However, some findings are clear. While most participating programs have a specific MT rotation, resident experience is quite varied. Many programs do not have a toxicologist on faculty. Future surveys will help to assess the development of MT education in EM residency programs. PMID- 11015280 TI - Emergency medicine resident related auto accidents-Is sleep deprivation a risk factor? AB - Published in: Acad Emerg Med. 2000; 7: 451. PMID- 11015281 TI - Low-flow perfusion in the heart following global ischemia improves LV function AB - Published in: Acad Emerg Med. 2000; 7: 508. PMID- 11015282 TI - Preliminary experience with an emergency department observation unit protocol for heart failure AB - Nearly all emergency department (ED) patients with congestive heart failure (CHF) are admitted to the hospital. Some of these patients might be best served by an explicit protocol for observation and treatment in the ED. OBJECTIVE: To describe the natural history of patients with CHF who are managed in an ED-based observation unit. METHODS: Patients presenting to the ED between 7/99 and 12/99 with symptoms of CHF were screened for eligibility. Patients with hemodynamic instability, respiratory failure, acute myocardial infarction, newonset heart failure, or severe complicating medical disease were excluded. Patients were managed in the observation unit for up to 12 hours before being admitted to the hospital. Clinical parameters including heart rate (HR), respiratory rate (RR), net diuresis, and global assessment of clinical severity (GACS) score, were charted. Patients were followed for 30 days to determine length of hospital stay (LOS), rates of morbidity, mortality, and readmission. RESULTS: Twenty-four patients (15 males, 9 females; median age = 58) were enrolled in the protocol. By 4 hr a reduction in HR and RR was seen in the majority of patients (19/24 and 13/24, respectively). By 6 hr the majority (13/24) had improvement in GACS, and by 12 hours nearly all (21/24) did. By 12 hr, most patients (13/24) had a net diuresis of >1500 mL. Upon completion of the protocol, inpatient length of stay was generally brief (median LOS = 1 day; range 1-8 days). At 30 days there were no deaths or adverse events observed. Four patients were readmitted to the hospital; 2 of these admissions were related to CHF. CONCLUSIONS: In this low risk group of patients with CHF, most showed clinical improvement within 6 to 12 hours after ED arrival. Further study is needed to determine whether an observation-unit protocol can safely and effectively reduce LOS for these patients. PMID- 11015283 TI - The penetrance of head injury management guidelines into the practice patterns of michigan emergency physicians AB - OBJECTIVE: The Brain Trauma Foundation published "Guidelines for the Management of Severe Head Injury" in 1996. These evidence based clinical guidelines (CGs) recommended against prophylactic hyperventilation and glucocorticoid use, and advocated for blood pressure (BP) resuscitation, and the careful use of mannitol in the event of elevated intracranial pressure (ICP). We sought to measure the penetrance of these guidelines into the practice patterns of Michigan emergency physicians (MEPs). METHODS: An anonymous mail survey was sent to all 566 MEPs who are members of the American College of Emergency Physicians. Three patients with severe head injury were presented, all with Glasgow Coma Scores (GCSs) of 8 or less. The physicians were asked to choose from 15 diagnostic and treatment options, which included: intubation and hyperventilation, BP resuscitation, IV mannitol administration, and IV glucocorticoid administration. RESULTS: 319 (56%) surveys were returned. 46% (95% CI = 40% to 51%) of MEPs elected to inappropriately use prophylactic hyperventilation, 78% (95% CI = 75% to 81%) did correct hypotension with systolic BP < 90 mm Hg. 83% (95% CI = 80% to 86%) administered mannitol appropriately. Very few MEPs administered IV glucocorticoids. CONCLUSIONS: Despite being published nearly 4 year ago, the "Guidelines for the Management of Severe Head Injury" have not been uniformly incorporated into the practice patterns of MEPs. Specifically, many physicians continue to rely on prophylactic hyperventilation despite strong recommendations against its use. PMID- 11015284 TI - Lung inflammation and function in a model of complement-mediated acute injury AB - Published by Acad Emerg Med. 2000; 7: 467-8. PMID- 11015285 TI - Resident procedure and resuscitation tracking using a palm computer AB - Resident procedure and resuscitation tracking is an onerous task required for residency accreditation and for future hospital privilege applications by the resident. To date, most tracking systems have been somewhat cumbersome and prone to data loss (forms not being filled out, recorded, etc.). Our residency program uses a palm computer database tracking system utilizing Palm III (3Com) hardware and a custom written data collection form utilizing an inexpensive, commercially available software package (Pendragon Forms (version 2), Pendragon Software Corporation, Libertyville, IL). Every resident receives a Palm III on entry into the residency. Residents enter basic demographic data and record procedures and resuscitations into the Palm III after each encounter. Generally, each patient logged requires approximately one minute for data entry. On a frequent basis, the resident's Palm is 'HotSync-ed' and the recorded data transferred to the program's central computer. Resident data are easily manipulated and reports are generated using a common, relational database program (Access97, Microsoft Corporation, Redmond, WA). We have found this system to be relatively inexpensive, to improve data capture, to reduce demands on secretarial time, and to allow improved tracking of resident procedure and resuscitation experiences. PMID- 11015286 TI - Caspase-1-inhibitor ac-YVAD-cmk reduces LPS-lethality in rats without affecting haematology or cytokine responses. AB - The effect of acetyl - tyrosyl-valyl-alanyl-aspartyl - chloromethylketone (ac YVAD-cmk), an irreversible caspase-1 (IL-1beta converting enzyme, ICE) inhibitor on mortality, leukocyte and platelet counts and cytokine levels was investigated in a double-blind rat model of endotoxaemia. Intravenous (i.v.) bolus administration of lipopolysaccharide (LPS) (25-75 mg kg(-1), n=12 per group) to anaesthetized rats induced a dose dependent increase in mortality over 8 h (LD(50)=48 mg kg(-1)). During this period, animals became leukopenic and thrombocytopenic. Serum levels of IL-beta, IL-6, and TNF-alpha were highly elevated. Pretreatment of rats with ac-YVAD-cmk at a dose of 12.5 micromol kg(-1) significantly reduced mortality from 83 to 33% using Log Rank analysis. However, ac-YVAD-cmk did not modify blood cell counts or cytokine profiles as compared with the LPS-drug vehicle group. These data lay credence to the potential importance of caspase-1-inhibition in modifying the inflammatory response to endotoxin. Further investigations are warranted in understanding the relationship between caspase-1 inhibition, cytokine production and animal survival in different experimental paradigms of sepsis. PMID- 11015287 TI - On the in vitro vasoactivity of bile acids. AB - We compared the vasorelaxant action of nine different bile acids and correlated their vasorelaxant activity with their individual indices for hydrophobicity or lipophilicity. Vasorelaxant activity correlated with the relative lipid solubility of bile acids with lipophilic bile acids exhibiting the greatest vasorelaxant activity with modest to no vasorelaxant activity exhibited by hydrophilic bile acids. We also investigated whether bile acid-induced vasorelaxation is mediated by antagonism of a prototypal contractile receptor, the alpha(1)-adrenoceptor, by stimulation of a bile acid surface membrane receptor, by the release of endothelium-derived relaxant factors, by promoting the generation of reactive oxygen species and increasing the extent of lipid peroxidation, or by modifying membrane fluidity. Lipophilic bile acids induce vasorelaxation possibly by antagonizing alpha(1)-adrenoceptors, a phenomenon that manifests itself as a lowering of the affinity of vascular alpha(1) adrenoceptors. Bile acid-induced vasorelaxation was not dependent upon stimulation of a bile acid surface membrane receptor or the release of endothelium-derived relaxant factors. Lipophilic bile acids can also increase the extent of lipid peroxidation with a subtle reduction in the fluidity of rat vascular smooth muscle membranes not associated with loss of membrane cholesterol or phospholipid. We have concluded that lipophilic bile acids are non-selective vasorelaxants whose mechanism of action is a multifaceted process involving antagonism of contractile surface membrane receptors possibly effected by an increased extent of lipid peroxidation and/or membrane fluidity but occurs independent of the release of endothelial-derived relaxant factors or stimulation of a surface membrane bile acid binding site. PMID- 11015288 TI - Cefaclor, a cephalosporin antibiotic, delays gastric emptying rate by a CCK-A receptor-mediated mechanism in the rat. AB - Studies in vitro suggest that cephalosporin antibiotics release the gut hormone cholecystokinin. Cholecystokinin is known to inhibit gastric emptying. Here we examine the effects of cefaclor on gastric emptying and intestinal motility. Male Sprague-Dawley rats were fitted with gastric cannulas. Following a 3-week recovery, the rate of gastric emptying of saline, peptone (4.5%) or cefaclor was determined after instillation into the gastric cannula, while intestinal transit was measured by using the propagation of arabic gum + charcoal mixture given intraduodenally. Gastric emptying of saline was significantly delayed by the addition of cefaclor (3, 10, 30 or 100 mM). The CCK-A antagonist SR-27897B (1 mg kg(-1), i.p.) reversed the delay induced by 10 mM cefaclor, whereas the CCK-B antagonist CI-988 (1 mg kg(-1), i.p.) had no significant effect. In capsaicin treated rats, 10 mM cefaclor emptied more rapidly than in vehicle-treated animals. Thirty-minute intestinal transit was increased at 30 and 100 mM of cefaclor, while the gastric acid secretion following cefaclor instillation was no different than the group which received saline. The cephalosporin antibiotic cefaclor appears to be a potent stimulant of CCK release from gut endocrine cells, resembling the effects of peptone. Cefaclor delays gastric emptying via capsaicin-sensitive afferent pathways, which involve CCK-A receptor interaction. PMID- 11015289 TI - RSD1019 suppresses ischaemia-induced monophasic action potential shortening and arrhythmias in anaesthetized rabbits. AB - The electrophysiological actions of lidocaine, tedisamil and RSD1019 were assessed on normal and ischaemic cardiac tissue using monophasic action potentials (MAPs) recorded from the epicardium of anaesthetized rabbits. Drug effects on ischaemia-induced arrhythmias were assessed simultaneously in the same rabbits. Lidocaine, infused at 2.5, 5 and 10 micromol kg(-1) min(-1) i.v., accelerated and worsened the electrophysiological derangement caused by ischaemia, had profibrillatory actions and reduced the time to the occurrence of ventricular fibrillation (VF) relative to controls. Tedisamil, infused at 0.063, 0.125 and 0.25 micromol kg(-1) min(-1) i.v., prolonged MAP duration at 90% repolarization (MAPD(90%)) before induction of ischaemia in a dose-related manner; however, this effect was not maintained 5 min after induction of ischaemia. Tedisamil had no significant antiarrhythmic actions over the dose range tested. RSD1019, infused at 2, 4 and 8 micromol kg(-1) min(-1) i.v., produced a small increase in MAPD(90%) before induction of ischaemia and only at the highest dose tested. In contrast to tedisamil, RSD1019 suppressed ischaemia induced MAP shortening assessed 5 min after induction of ischaemia. This effect was dose-related. RSD1019 completely prevented ischaemia-induced tachyarrhythmias at the mid and highest infusion levels tested. The results of this study illustrate a pathologically targeted approach for preventing ischaemia-induced arrhythmias. Suppression of ischaemia-induced MAP shortening, demonstrated herein for RSD1019, represents a novel antifibrillatory approach. PMID- 11015290 TI - The involvement of cytokines in the second window of ischaemic preconditioning. AB - We utilized a rat model of myocardial infarction to investigate whether manganese superoxide dismutase (Mn-SOD), an intrinsic radical scavenger, and tumour necrosis factor- alpha (TNF-alpha) and/or interleukin-1beta (IL-1beta) are involved in the late phase of ischaemic preconditioning (IP). IP was induced in anaesthetized rats by four 3-min left coronary artery (LCA) occlusions, each separated by 10 min of reperfusion. Twenty-four hours after the repetitive brief ischaemia, the LCA was occluded for 20 min followed by reperfusion for 48 h. IP reduced the infarct size by approximately 46% as determined after 48 h of reperfusion. Antisense oligodeoxynucleotides to Mn-SOD inhibited the increases in Mn-SOD content and activity, and abolished the expected decrease in myocardial infarct size. Sense or scrambled oligodeoxynucleotides did not abolish either Mn SOD induction or tolerance to ischaemia/reperfusion. The simultaneous administration of the antibodies to TNF-alpha (0.5 ml) and IL-1beta (0.5 mg) prior to IP abolished the cardioprotection and the increase in Mn-SOD activity induced by IP. We conclude that the induction and activation of Mn-SOD, mediated by TNF-alpha and IL-1beta after IP, plays an important role in the acquisition of late-phase cardioprotection against ischaemia/reperfusion injury in rats. PMID- 11015291 TI - Gene expression of receptors and enzymes involved in GABAergic and glutamatergic neurotransmission in the CNS of rats behaviourally dependent on ethanol. AB - The steady state levels of the messenger RNA (mRNA) of eight GABA(A) receptor subunits, five glutamate receptor subunits and seven enzymes involved in the synthesis of glutamate and GABA were measured in eight regions of rat brain in a recently developed animal model of 'behavioural dependence' on ethanol. 'Behavioural dependence' including loss of control was induced by offering the rats the choice between ethanol and water over a 9-month period (Group A). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet 'behaviourally dependent', Group B), a group forced to consume ethanol as sole fluid over a 9-month period (also not 'behaviourally dependent', Group C) and ethanol-naive control rats (Group D). All groups were sacrificed 1 month after the ethanol was withdrawn. The mRNA concentrations of all eight GABA receptor subunits, four out of the five subunits of different glutamate receptors and those of seven enzymes involved in GABA and glutamate production were reduced almost exclusively in the parieto-occipital cortex in Groups A and B, but not Group C. These data suggest that the synthesis of glutamate and GABA and the activities of their respective neurons are selectively impaired in the parieto-occipital cortex in the groups having consumed ethanol in a free-choice design, in which its rewarding properties can better take effect than after forced administration. As the parieto-occipital cortex is believed to contain emotional memory structures, it may be hypothesized that the glutamatergic and GABAergic neuronal systems in this area are involved in the development of memory for reward from ethanol. However, they are not specifically associated with 'behavioural dependence'. PMID- 11015293 TI - Orphan-receptor ligand human urotensin II: receptor localization in human tissues and comparison of vasoconstrictor responses with endothelin-1. AB - We have determined the distribution of receptors for human urotensin-II (U-II) in human and rat CNS and peripheral tissues. In rat, [(125)I]-U-II binding density was highest in the abducens nucleus of brainstem (139.6+/-14 amol mm(-2)). Moderate levels were detected in dorsal horn of spinal cord and lower levels in aorta (22. 5+/-6 amol mm(-2)). In human tissues density was highest in skeletal muscle and cerebral cortex ( approximately 30 amol mm(-2)), with lower levels (<15 amol mm(-2)) in kidney cortex and left ventricle. Little binding was identified in atria, conducting system of the heart and lung parenchyma. Receptor density was less in human coronary artery smooth muscle (14.6+/-3 amol mm(-2), n=10) than rat aorta with no significant difference between normal and atherosclerotic vessels. In human skeletal muscle [(125)I]-U-II bound to a single receptor population with K(D)=0.24+/-0.17 nM and B(max)=1.97+/-1.1 fmol mg(-1) protein (n=4). U-II contracted human coronary, mammary and radial arteries, saphenous and umbilical veins with sub-nanomolar EC(50) values. U-II was 50 times more potent in arteries and <10 times more potent in veins than endothelin-1 (ET 1). The maximum response to U-II ( approximately 20% of control KCl) was significantly less than to ET-1 ( approximately 80% KCl). In contrast, in rat aorta, U-II and ET-1 were equipotent with similar maximum responses. This is the first report of high affinity receptors for [(125)I]-U-II in human CNS and peripheral tissues. This peptide produces potent, low efficacy, vasoconstriction in human arteries and veins. These data suggest a potential role for U-II in human physiology. PMID- 11015292 TI - Potent stimulation and inhibition of the CFTR Cl(-) current by phloxine B. AB - The effects of the fluoresceine derivative, phloxine B, on the Cl(-) current through the cystic fibrosis transmembrane conductance regulator (CFTR) were examined in Xenopus oocytes expressing human CFTR. In whole oocytes, the CFTR Cl( ) current (I(CFTR)) was activated by superfusion with isobutylmethylxanthine and forskolin. I(CFTR) was stable during activation and deactivated rapidly upon washout of the activation solution. Phloxine B slowed deactivation and, at high concentrations, inhibited I(CFTR) weakly. In excised inside-out macropatches, I(CFTR) was activated by the catalytic subunit of protein kinase A (cPKA) and MgATP. Phloxine B (0.01 - 3 microM), applied after activation, increased I(CFTR) within 30 s followed by a slow decrease which became dominant at high concentrations. Slowing of deactivation of the CFTR was observed at all concentrations. The effect of phloxine B after 30 s had a bell-shaped concentration-dependence with midpoints at 45 and 1600 nM for the stimulatory and the inhibitory limb, respectively; maximum stimulation was about 1.8 times. The slow inhibitory component, measured after 6 min, occurred with an IC(50) value of approximately 1 microM. In the absence of cPKA, phloxine B did not stimulate I(CFTR). In the presence of cPKA and MgATP, the effects of phloxine B were more prominent at low (0.02 mM) than at high ATP (2 mM). The data show that phloxine B modulates I(CFTR) by increasing channel activity and slowing channel deactivation; at high concentrations inhibition dominates. The effects may be mediated by direct interactions with CFTR from the inside of the cell. PMID- 11015294 TI - Inhibition of acetylcholine muscarinic M(1) receptor function by the M(1) selective ligand muscarinic toxin 7 (MT-7). AB - MT-7 (1 - 30 nM), a peptide toxin isolated from the venom of the green mamba Dendroaspis angusticeps and previously found to bind selectively to the muscarinic M(1) receptor, inhibited the acetylcholine (ACh)-stimulated [(35)S] guanosine-5'-O-(3-thio)triphosphate ([(35)S]-GTPgammaS) binding to membranes of Chinese hamster ovary (CHO) cells stably expressing the cloned human muscarinic M(1) receptor subtype. MT-7 failed to affect the ACh-stimulated [(35)S]-GTPgammaS binding in membranes of CHO cells expressing either the M(2), M(3) or M(4) receptor subtype. In N1E-115 neuroblastoma cells endogenously expressing the M(1) and M(4) receptor subtypes, MT-7 (0.3 - 3.0 nM) inhibited the carbachol (CCh) stimulated inositol phosphates accumulation, but failed to affect the CCh-induced inhibition of pituitary adenylate cyclase activating polypeptide (PACAP) 38 stimulated cyclic AMP accumulation. In both CHO/M(1) and N1E-115 cells the MT-7 inhibition consisted in a decrease of the maximal agonist effect with minimal changes in the agonist EC(50) value. In CHO/M(1) cell membranes, MT-7 (0.05 - 25 nM) reduced the specific binding of 0.05, 1.0 and 15 nM [(3)H]-N methylscopolamine ([(3)H]-NMS) in a concentration-dependent manner, but failed to cause a complete displacement of the radioligand. Moreover, MT-7 (3 nM) decreased the dissociation rate of [(3)H]-NMS by about 5 fold. CHO/M(1) cell membranes preincubated with MT-7 (10 nM) and washed by centrifugation and resuspension did not recover control [(3)H]-NMS binding for at least 8 h at 30 degrees C. It is concluded that MT-7 acts as a selective noncompetitive antagonist of the muscarinic M(1) receptors by binding stably to an allosteric site. PMID- 11015295 TI - Developmental regulation of hippocampal excitatory synaptic transmission by metabotropic glutamate receptors. AB - The aims of this study were, to use agonists selective for the 3 mGlu receptor groups to identify developmental changes in their effects, and to assess the usefulness of proposed selective antagonists as pharmacological tools. Hippocampal slices (400 microm) were prepared from neonate (9 - 14 days) and young adult (5 - 7 weeks) Sprague-Dawley rats. Field excitatory postsynaptic potentials (fEPSP) were recorded from CA1. DHPG (100 microM), a group I agonist, produced a slowly developing enhancement of fEPSP slope in slices from adults. In slices from neonates, DHPG (75 microM) depressed fEPSP slope. DCG-IV (500 nM), a group II agonist, did not affect the fEPSP recorded from slices from adults whereas perfusion in neonate slices produced a sustained depression. The group III agonist L-AP4 (50 microM) was ineffective in adult slices but depressed fEPSP slope in slices prepared from neonates. DHPG-induced depression of fEPSP slope was inhibited by 4-CPG (400 microM), a group I antagonist, but was unaffected by MCCG (500 microM) and MAP4 (500 microM), group II and III receptor antagonists respectively. MCCG but not MAP4 antagonized the effects of DCG-IV with 4-CPG producing variable effects. The effect of L-AP4 was unaffected by MCCG, blocked by MAP4, and enhanced by 4-CPG. The results show that the effects of the agonists for all groups of mGlu receptors are developmentally regulated. Furthermore, MCCG and MAP4 behave as effective and selective antagonists for group II and group III mGlu receptors respectively, whereas the usefulness of 4-CPG as a group I antagonist may be limited. PMID- 11015296 TI - Oncostatin M synergises with house dust mite proteases to induce the production of PGE(2) from cultured lung epithelial cells. AB - The release of PGE(2) and nitric oxide (NO) from the respiratory epithelium may act to dampen inflammation. In other tissues, oncostatin M (OSM), a potent inducer of epithelial antiproteases, has also been shown to interact with IL 1beta to stimulate PGE(2) release. However, whether OSM interacts with pro inflammatory cytokines and proteases in the production of anti-inflammatory eicosanoids and NO from airway epithelium is unknown. The effect of OSM and the related cytokine leukaemia inhibitory factor (LIF) on PGE(2) and NO production by the respiratory epithelial cell line, A549 in response to pro-inflammatory cytokines as well as protease-rich house dust mite (HDM) fractions and a protease deficient rye grass pollen extract was examined by immunohistochemistry, cell culture, ELISA and enzyme-immunoassay. Cells treated with a mixture of IL-1beta, IFNgamma and LPS for 48 h produced a 9 fold increase in PGE(2) and a 3 fold increase in NO levels (both P<0.05). Both OSM and LIF were without effect. However, OSM added together with the cytokine mixture synergistically enhanced PGE(2) production (22 fold, P<0.05). OSM also synergistically enhanced PGE(2) production in response to a cysteine protease-enriched, but not serine protease enriched HDM fraction (P<0.05). Rye grass extract, neither alone nor in combination with OSM, induced PGE(2) or NO production, although it did induce the release of GM-CSF. These observations suggest that OSM is an important co-factor in the release of PGE(2) and NO from respiratory epithelial cells and may play a role in defense against exogenous proteases such as those derived from HDM. PMID- 11015297 TI - Amphotericin B severely affects expression and activity of the endothelial constitutive nitric oxide synthase involving altered mRNA stability. AB - The therapeutic use of the antifungal drug amphotericin B (AmB) is limited due to severe side effects like glomerular vasoconstriction and risk of renal failure during AmB administration. As nitric oxide (NO) has substantial functions in renal autoregulation, we have determined the effects of AmB on endothelial constitutive NO synthase (ecNOS) expression and activity in human and rat endothelial cell cultures. AmB used at concentrations of 0.6 to 1.25 microg ml( 1) led to increases in ecNOS mRNA and protein expression as well as NO production. This was the result of an increased ecNOS mRNA half-life. In contrast, incubation of cells with higher albeit subtoxic concentrations of AmB (2.5 - 5.0 microg ml(-1)) resulted in a decrease or respectively in completely abolished ecNOS mRNA and protein expression with a strongly reduced or inhibited ecNOS activity, due to a decrease of ecNOS mRNA half-life. None of the AmB concentrations affected promoter activity as found with a reporter gene construct stably transfected into ECV304 cells. Thus, our experiments show a concentration dependent biphasic effect of AmB on expression and activity of ecNOS, an effect best explained by AmB influencing ecNOS mRNA stability. In view of the known renal accumulation of this drug the results reported here could help to elucidate its renal toxicity. PMID- 11015298 TI - Histamine H(2) receptors mediate the inhibitory effect of histamine on human eosinophil degranulation. AB - The effect of histamine on human eosinophil degranulation and the receptor mediating such effect were studied in vitro using the complement C5a-mediated eosinophil peroxidase (EPO) release model. Following pre-treatment with 5 microg ml(-1) cytochalasin B(CB), C5a induced a concentration-dependent release of EPO from eosinophils isolated from healthy donors. Histamine (0.1-50 microM), but not L-histidine, inhibited concentration-dependently C5a-induced EPO release with IC(50) (95% CI) of 0.6 microM (0.3-1.2 microM) and maximal inhibition of approximately 60%. A similar effect was seen with the selective H(2) agonists dimaprit (IC(50) (95% CI)=6.9 microM (3.2-10.6 microM)) and amthamine (IC(50) (95% CI)=0.4 microM (0.2-0.7 microM)). Neither the selective H(1) agonist 6-(2-(4 imidazolyl)ethylamino)-N-(4-trifluoromethylphenyl) heptanecarboxamide(HTMT), nor the selective H(3) agonists imetit (up to 100 microM) had any significant effect. The inhibition by histamine was reversed by cimetidine (0.1-30 microM) and other H(2) antagonists, but not the H(1) antagonist mepyramine (1.0- 100 microM), nor the H(3) antagonist thioperamide (1.0-100 microM). Cimetidine (1-30 microM) shifted to the right the dimaprit log dose-response curve, producing a pA(2) value of 5.9 and Schild's plot slope of 0.98, thus confirming simple competitive antagonism. Histamine (10-100 microM) increased intracellular level of adenosine 3',5'-cyclic monophosphate, which was completely abolished by cimetidine (30 microM), but not mepyramine or thioperamide. The cyclic AMP analogue - dibutyryl cyclic AMP - also inhibited degranulation (IC(50) approximately 300 microM). The cyclic AMP phosphodiesterase(PDE) IV inhibitor rolipram (10 microM) synergistically enhanced the inhibition of EPO release by histamine. These results suggest that histamine, via stimulation of H(2) receptors and a consequent elevation of intracellular levels of cyclic AMP, inhibits human eosinophil degranulation. PMID- 11015299 TI - Differential regulation of P2Y(11) receptor-mediated signalling to phospholipase C and adenylyl cyclase by protein kinase C in HL-60 promyelocytes. AB - The regulatory mode of the P2Y(11) purinoceptor-mediated signalling cascades towards phospholipase C and adenylyl cyclase was studied in HL-60 promyelocytes. Treatment with the potent P2Y(11) receptor activator dATP evoked an elevated intracellular Ca(2+) concentration ([Ca(2+)](i)) and inositol 1,4,5-trisphosphate (IP(3)) production that was sustained for longer than 30 min. However, the dATP induced responses were significantly inhibited by the activation of protein kinase C after a short exposure to phorbol 12-myristate 13-acetate (PMA). dATP also potently stimulated cyclic AMP production with half maximum effect seen at 23+/-7 microM dATP. In addition, a 5-min pretreatment with PMA enhanced the dATP stimulated cyclic AMP accumulation. PMA potentiated the cyclic AMP production when adenylyl cyclase was activated directly by forskolin or indirectly by G protein activation after cholera toxin treatment. dATP also enhanced the forskolin-mediated cyclic AMP generation. Treatment of the cells with 10 microM U 73122, which almost completely blocked the dATP-stimulated IP(3) production and [Ca(2+)](i) rise, had no effect on cyclic AMP accumulation, while 10 microM 9 (tetrahydro-2-furyl)adenine (SQ 22536), which inhibited the adenylyl cyclase activation, did not effect the dATP-stimulated phosphoinositide turnover. Taken together, the results indicate that P2Y(11) receptor-mediated activation of phospholipase C and adenylyl cyclase occurs through independent pathways and is differentially regulated by protein kinase C in HL-60 cells. PMID- 11015300 TI - Differences in the pharmacological properties of rat and chicken brain fatty acid amidohydrolase. AB - The pharmacological properties of fatty acid amidohydrolase (FAAH) were investigated in brains of 35-day-old chickens, since nothing is known about the enzyme in avian species. FAAH activity towards both [(3)H]-palmitoylethanolamide (PEA) [K(M)=1.5 microM] and [(3)H]-anandamide (AEA) [K(M)=5.4 microM] was demonstrated in the chicken brains. The chicken FAAH was inhibited by the substrate analogues oleyl trifluoromethylketone (OTMK) and diazomethylarachidonyl ketone (DAK) with similar potencies to the rat FAAH. However, in contrast to the rat brain, phenylmethylsulphonyl fluoride (PMSF) and the enantiomers of ibuprofen had very weak effects on chicken brain FAAH. Indomethacin and niflumic acid were found to inhibit rat brain AEA hydrolysis. The inhibition by indomethacin was reversible and competitive, with a K(i) value of 120 microM. Chicken FAAH was less sensitive to indomethacin than its rodent counterpart, but the inhibition was also competitive (K(i)). It is concluded that chicken FAAH activity has different pharmacological properties to its rodent counterpart. PMID- 11015301 TI - The effects of caffeine on ATP-sensitive K(+) channels in smooth muscle cells from pig urethra. AB - The effects of caffeine on both levcromakalim-induced macroscopic and unitary currents in pig proximal urethra were investigated by the use of patch-clamp techniques (conventional whole-cell configuration and cell-attached configuration). The effects of caffeine were also examined on currents in inside out patches of COS7 cells expressing carboxy terminus truncated inwardly rectifying K(+) channel (Kir6.2) subunits (i.e. Kir6.2DeltaC36) which form ATP sensitive K(+) channels (K(ATP) channels). In conventional whole-cell configuration, the levcromakalim (100 microM)-induced inward current (symmetrical 140 mM K(+) conditions) was inhibited by caffeine (> or =1 mM) at a holding potential of -50 mV. In contrast, ryanodine (10 microM) caused no significant inhibitory effect on the gradual decay of the levcromakalim-induced current at 50 mV. The amplitude of the 30 microM levcromakalim-induced current was enhanced by 3-isobutyl-1-methylxanthine (IBMX, 100 microM). In cell-attached configuration, the levcromakalim-induced K(+) channel openings were inhibited by subsequent application of 10 mM caffeine, decreasing the channel open probability at -50 mV. Reverse transcriptase-polymerase chain reaction (RT - PCR) analysis revealed the presence of Kir6.2 transcript in pig urethra. Caffeine (> or =3 mM) inhibited the channel activity of Kir6.2DeltaC36 expressed in COS7 cells (3 mM caffeine, 65+/-6%, n=4; 10 mM caffeine, 29+/-2%, n=4). These results suggest that caffeine can inhibit the activity of K(ATP) channels through a direct blocking effect on the pore-forming Kir subunit. PMID- 11015302 TI - Advantages of heterologous expression of human D2long dopamine receptors in human neuroblastoma SH-SY5Y over human embryonic kidney 293 cells. AB - The human D2long dopamine receptor when expressed heterologously in a human neuronal cell line, SH-SY5Y, produced more robust functional signals than when expressed in a human embryonic kidney cell line, HEK293. Quinpirole (agonist) induced GTPgamma(35)S binding and high affinity sites were 3 - 4 fold greater in SH-SY5Y than in HEK293 cells. N-type Ca(2+) channel currents present in SH-SY5Y cells, but not HEK293 cells, were inhibited potently by quinpirole with a half maximal inhibitory concentration of 0.15+/-0.03 nM. Inhibition of adenylyl cyclases by agonists, on the other hand, was of similar potency and efficacy in the two cell lines. GTPgamma(35)S-Bound Galpha subunits from quinpirole-activated and solubilized membranes were monitored upon immobilization with various Galpha specific antibodies. Galpha(i) and Galpha(o) subunits were highly labelled with GTPgamma(35)S in SH-SY5Y cells, but only Galpha(i) subunits were labelled in HEK293 cells. The additional G(o) coupling in SH-SY5Y cells could arise, at least in part, from the presence of G(o) coupled-effectors, such as the N-type Ca(2+) channel, and may contribute to robust agonist-induced GTPgamma(35)S binding, which is a reliable means for measuring ligand intrinsic efficacy. It appears that expression of neuronal G protein-coupled receptors in neuronal environments could reveal additional functional characteristics that are absent in non neuronal cell lines. This appears to be due to, at least in part, to the presence of neuron-specific effectors. These findings underscore the importance of the cellular environment in which drug actions are examined, particularly in the face of intensive efforts to develop drugs for G protein-coupled receptors of various origins. PMID- 11015303 TI - Nitric oxide and sodium nitroprusside-induced relaxation of the human umbilical artery. AB - In the human umbilical artery (HUA) pre-contracted with the thromboxane mimetic U46619 or with 5-hydroxytryptamine (5-HT), (and pretreated with indomethacin (3 microM) to suppress the synthesis of prostanoids), authentic nitric oxide (NO) evoked concentration-dependent relaxation (pEC(50) 7.05 and 5.99, respectively). In contrast, sodium nitroprusside (SNP) induced relaxation only in U46619 pre contracted HUA (pEC(50) 6.52). At high (>300 mmHg) vs low (<55 mmHg) oxygen tension the dose-response curves to NO- and SNP-induced relaxations were biphasic and shifted leftward. Preincubation of the arterial rings with the soluble guanylyl cyclase (sGC) inhibitor 1H[1,2,4]oxadiazolo[4, 3-a]quinoxalin-1-one (ODQ; 10 microM) shifted the concentration-response curve to NO, reduced the maximal relaxation response to NO (E(max) 71%) and to SNP (E(max) 10%). Pre exposure of HUA rings to high extracellular K(+) (50 mM) reduced E(max) relaxation responses to NO (36%) and SNP (1%). Pretreatment of the HUA with the K(+) channel inhibitors, tetraethylammonium (TEA, 1 mM), 4-aminopyridine (4-AP, 0.5 mM), charybdotoxin (0.1 microM) or iberiotoxin (0.1 microM) increased the pEC(30) for NO and SNP and changed the shape of the dose-response curves from biphasic to monophasic. Pre-incubation of HUA rings with TEA (1 mM), 4-AP (0.5 mM) and ODQ (10 microM) significantly reduced the NO-induced maximal relaxation (E(max) 26%) but not the pEC(50) (5.60). These data indicate that SNP-induced relaxation in the HUA is primarily mediated via sGC-cyclic GMP whereas NO-induced relaxation also involves the activation of K(V) and K(Ca) channels and a cyclic GMP/K(+) channel-independent mechanism(s). PMID- 11015304 TI - Effects of tetrandrine on calcium transport, protein fluorescences and membrane fluidity of sarcoplasmic reticulum. AB - To understand whether the molecular mechanism of Tetrandrine (Tet)'s pharmacological effects is concerned with sarcoplasmic reticulum calcium transport so as to be involved in myocardial contractility, we observed the effects of Tet on calcium transport and membrane structure of rabbit skeletal muscle sarcoplasmic reticulum vesicles (SR) and rat cardiac sarcoplasmic reticulum vesicles (CSR). Calcium uptake was monitored with a dual-wavelength spectrophotometer. Protein conformation and fluorescence polarization were measured by fluospectrophotometric method and membrane lipids labelled with fluorescence probes for SR, respectively. 128 micromol l(-1) Tet reduced the initial rate of calcium uptake to 59% of control 6 min after reaction. Tet un competitively inhibited SR Ca(2+), Mg(2+)-ATPase activity, causing the stoichiometric ratio of SR Ca(2+)/ATP to decrease to 1.43 from 2.0 of control. Inhibitory rates on SR Ca(2+),Mg(2+)-ATPase by Tet were reduced from 60% in the absence of phosphate to 50% in the presence of phosphate and reduced from 92% in 1 mmol l(-1) ATP to 60% in 5 mmol l(-1) ATP. Tet markedly reduced SR intrinsic protein fluorescence, while it slightly decreased the thiol(SH)-modified protein fluorescence of SR labelled with N-(3-pyrene)-maleimide. Tet slightly increased fluorescence polarization in the middle and deep layers of SR membrane lipids labelled with 7- or 12-(9-anthroyloxy) stearic acid (AS) probes, whereas it did not change that of SR labelled with 1, 6-diphenyl-1,3,5-hexatrine (DPH). These results revealed that prevention of SR calcium uptake by Tet was due to inhibition of the SR calcium pump Ca(2+),Mg(2+)-ATPase, changes in spatial conformation of the pumps protein molecules and a decrease in the extent of motion of membrane lipid molecules, thus altering the regulation of [Ca(2+)](i) and myocardial contractility. PMID- 11015305 TI - Influence of beta-adrenoceptor tone on the cardioprotective efficacy of adenosine A(1) receptor activation in isolated working rat hearts. AB - This study investigated the role of beta-adrenoceptors in the cardioprotective and metabolic actions of adenosine A(1) receptor stimulation. Isolated paced (300 beats min(-1)) working rat hearts were perfused with Krebs-Henseleit solution containing 1.2 mM palmitate. Left ventricular minute work (LV work), O(2) consumption and rates of glycolysis and glucose oxidation were measured during reperfusion (30 min) following global ischaemia (30 min) as well as during aerobic conditions. Relative to untreated hearts, N(6)-cyclohexyladenosine (CHA, 0.5 microM) improved post-ischaemic LV work (158%) and reduced glycolysis and proton production (53 and 42%, respectively). CHA+propranolol (1 microM) had similar beneficial effects, while propranolol alone did not affect post-ischaemic LV work or glucose metabolism. Isoprenaline (10 nM) impaired post-ischaemic function and after 25 min ischaemia recovery was comparable with 30 min ischaemia in untreated hearts (41 and 53%, respectively). Relative to isoprenaline alone, CHA+isoprenaline improved recovery of LV work (181%) and reduced glycolysis and proton production (64 and 60%, respectively). In aerobic hearts, CHA, propranolol or CHA+propranolol had no effect on LV work or glucose oxidation. Glycolysis was inhibited by CHA, propranolol and CHA+propranolol (50, 53 and 52%, respectively). Isoprenaline-induced increases in heart rate, glycolysis and proton production were attenuated by CHA (85, 57 and 53%, respectively). The cardioprotective efficacy of CHA was unaffected by antagonism or activation of beta-adrenoceptors. Thus, the mechanism of protection by adenosine A(1) receptor activation does not involve functional antagonism of beta-adrenoceptors. PMID- 11015306 TI - Inverse agonism and neutral antagonism at a constitutively active alpha-1a adrenoceptor. AB - We have studied the antagonist action of prazosin and KMD-3213 in a constitutively active mutant of the human alpha-1a adrenoceptor in which Ala(271) was substituted to Thr and was expressed in CHO cells. Inverse agonism was characterized by up-regulation of receptor density, a decrease in basal GTPgammaS binding, and a reduction in basal inositol-1,4,5-trisphosphate (IP(3)) level. According to the above criteria, prazosin acted as an inverse agonist, whilst KMD 3213 behaved as a neutral antagonist. Compared with the wild-type receptor, mutant receptor exhibited single affinity sites for [(3)H]-prazosin, [(3)H]-KMD and the non-radioactive ligands tested, and displayed significantly higher affinities for several agonists but not for the two antagonists. Administration of KMD-3213 to prazosin-treated CHO cells expressing the mutant receptor reversed the inverse agonism of prazosin resulting in rapid increases in cellular IP(3), in intracellular [Ca(2+)] and in the rate of extracellular acidification. These results indicated that a neutral antagonist can reverse the action of an inverse agonist at the receptor site. The distinct properties of inverse agonist and neutral antagonist in affecting receptor function may be important for the clinical use of such antagonists. PMID- 11015307 TI - Inhibition of small G proteins of the rho family by statins or clostridium difficile toxin B enhances cytokine-mediated induction of NO synthase II. AB - In order to investigate the involvement of Ras and/or Rho proteins in the induction of the inducible isoform of nitric oxide synthase (NOS II) we used HMG CoA reductase inhibitors (statins) and Clostridium difficile toxin B (TcdB) as pharmacological tools. Statins indirectly inhibit small G proteins by preventing their essential farnesylation (Ras) and/or geranylgeranylation (Rho). In contrast, TcdB is a glucosyltransferase and inactivates Rho-proteins directly. Human A549/8- and DLD-1 cells as well as murine 3T3 fibroblasts were preincubated for 18 h with statins (1 - 100 microM) or TcdB (0.01-10 ng ml(-1)). Then NOS II expression was induced by cytokines. NOS II mRNA was measured after 4 - 8 h by RNase protection assay, and NO production were measured by the Griess assay after 24 h. Statins and TcdB markedly increased cytokine-induced NOS II mRNA expression and NO production. Statin-mediated enhancement of NOS II mRNA expression was reversed almost completely by cotreatment with mevalonate or geranylgeranylpyrophosphate. It was only slightly reduced by farnesylpyrophosphate. Therefore, small G proteins of the Rho family are likely to be involved in NOS II induction. In A549/8 cells stably transfected with a luciferase reporter gene under the control of a 16 kb fragment of the human NOS II promoter (pNOS2(16)Luc), statins produced only a small increase in cytokine induced NOS II promoter activity. In contrast, statins had a considerable superinducing effect in DLD-1 cells stably transfected with pNOS2(16)Luc. In conclusion, our studies provide evidence that statins and TcdB potentiate cytokine-induced NOS II expression via inhibition of small G proteins of the Rho family. This in turn results in an enhanced NOS II promoter activity and/or a prolonged NOS II mRNA stability. PMID- 11015308 TI - Contractile responses to sumatriptan and ergotamine in the rabbit saphenous vein: effect of selective 5-HT(1F) receptor agonists and PGF(2alpha). AB - Contractile responses to ergotamine, sumatriptan and the novel 5-HT(1F) receptor agonists, LY334370 and LY344864 were examined using the rabbit saphenous vein. Ergotamine (pEC(50)=8.7+/-0.06) was 30 fold more potent than 5-hydroxytryptamine (5-HT) (pEC(50)=7.2+/-0.13) and 300 fold more potent than sumatriptan (pEC(50)=6.0+/-0.08) in contracting the rabbit saphenous vein in vitro. The selective 5-HT(1F) receptor agonists, LY334370 or LY344864 (up to 10(-4) M), did not contract the rabbit saphenous vein. The contractile response to ergotamine in this tissue resulted from activation of both alpha(1) and 5-HT(1B/1D) receptors based on the observation that prazosin (10(-6) M), an alpha-adrenoceptor antagonist, and GR127935 (10(-8) M) a 5-HT(1B/1D) receptor antagonist, dextrally shifted the contractile response to ergotamine. In contrast, prazosin (10(-6) M) did not alter contraction to sumatriptan whereas GR127935 (10(-8) M) was a potent antagonist (-log K(B)=10.0) suggesting that sumatriptan-induced contraction of the rabbit saphenous vein was mediated only by activation of receptors similar or identical to 5-HT(1B/1D) receptors. PGF(2alpha) (3x10(-7) M) produced a modest increase (approximately 5.0 - 10.0% maximum PGF(2alpha) contraction) in saphenous vein force. Precontraction with PGF(2alpha) (3x10(-7) M) dramatically augmented the potency and maximal contractile response to sumatriptan (pEC(50)=7.1) and modestly enhanced the contractile potency of ergotamine (pEC(50)=9.0) in the rabbit saphenous vein. However, PGF(2alpha) (3x10(-7) M) only unmasked a contraction to the 5-HT(1F) receptor agonists when concentrations exceeded 10(-5) M, concentrations considerably higher than their 5-HT(1F) receptor affinities. LY334370 (10(-6) M) pretreatment did not alter contraction to either sumatriptan or ergotamine and a higher concentration (10(-5) M) of LY334370 or LY344864 inhibited contraction to sumatriptan. Thus, activation of 5-HT(1F) receptors will not induce vascular contraction (either alone or following modest tone with PGF(2alpha)) or augment contraction to other contractile agonists in the rabbit saphenous vein. PMID- 11015309 TI - The bee venom peptide tertiapin underlines the role of I(KACh) in acetylcholine induced atrioventricular blocks. AB - Acetylcholine (ACh) is an important neuromodulator of cardiac function that is released upon stimulation of the vagus nerve. Despite numerous reports on activation of I(KACh) by acetylcholine in cardiomyocytes, it has yet to be demonstrated what role this channel plays in cardiac conduction. We studied the effect of tertiapin, a bee venom peptide blocking I(KACh), to evaluate the role of I(KACh) in Langendorff preparations challenged with ACh. ACh (0.5 microM) reproducibly and reversibly induced complete atrioventricular (AV) blocks in retroperfused guinea-pig isolated hearts (n=12). Tertiapin (10 to 300 nM) dose dependently and reversibly prevented the AV conduction decrements and the complete blocks in unpaced hearts (n=8, P<0.01). Tertiapin dose-dependently blunted the ACh-induced negative chronotropic response from an ACh-induced decrease in heart rate of 39+/-16% in control conditions to 3+/-3% after 300 nM tertiapin (P=0.01). These effects were not accompanied by any significant change in QT intervals. Tertiapin blocked I(KACh) with an IC(50) of 30+/-4 nM with no significant effect on the major currents classically associated with cardiac repolarisation process (I(Kr), I(Ks), I(to1), I:(sus), I(K1) or I(KATP)) or AV conduction (I(Na) and I(Ca(L))). In summary, tertiapin prevents dose-dependently ACh-induced AV blocks in mammalian hearts by inhibiting I(KACh). PMID- 11015310 TI - Dual modulation by thrombin of the motility of rat oesophageal muscularis mucosae via two distinct protease-activated receptors (PARs): a novel role for PAR-4 as opposed to PAR-1. AB - Since protease-activated receptors (PARs) are distributed throughout the gastrointestinal tract, we investigated the role of PARs in modulation of the motility of the rat oesophageal muscularis mucosae. Thrombin produced contraction of segments of the upper and lower part of the smooth muscle. Trypsin contracted both the muscle preparations only at high concentrations. SFLLR-NH(2) and TFLLR NH(2) (PAR-1-activating peptides), but not the PAR-1-inactive peptide FSLLR NH(2), evoked a marked contraction. In contrast, the PAR-2 agonist SLIGRL-NH(2) and the PAR-4 agonist GYPGKF-NH(2) caused no or only a negligible contraction. In oesophageal preparations precontracted with carbachol, thrombin produced a dual action i.e. relaxation followed by contraction. TFLLR-NH(2) further contracted the precontracted preparations with no preceding relaxation. GYPGKF-NH(2), but not the inactive peptide GAPGKF-NH(2), produced marked relaxation. Trypsin or SLIGRL-NH(2) caused no relaxation. The PAR-1-mediated contraction was completely abolished in Ca(2+)-free medium and considerably attenuated by nifedipine (1 microM) and in a low Na(+) medium. The PAR-4-mediated relaxation was resistant to tetrodotoxin (10 microM), apamin (0.1 microM), charybdotoxin (0.1 microM), L-N(G) nitroarginine methyl ester (100 microM), indomethacin (3 microM), propranolol (5 microM) or adenosine 3', 5'-cyclic monophosphorothioate, 8-bromo, Rp-isomer (30 microM). Thus, thrombin plays a dual role in modulating the motility of the oesophageal muscularis mucosae, producing contraction via PAR-1 and relaxation via PAR-4. The PAR-1-mediated effect appears to occur largely through increased Na(+) permeability followed by activation of L-type Ca(2+) channels and subsequent influx of extracellular Ca(2+). Our data could provide evidence for a novel role of PAR-4 as opposed to PAR-1, although the underlying mechanisms are still open to question. PMID- 11015311 TI - Prodigiosin from the supernatant of Serratia marcescens induces apoptosis in haematopoietic cancer cell lines. AB - The effects of supernatant from the bacterial strain Serratia marcescens 2170 (CS 2170) on the viability of different haematopoietic cancer cell lines (Jurkat, NSO, HL-60 and Ramos) and nonmalignant cells (NIH-3T3 and MDCK) was studied. We examined whether this cytotoxic effect was due to apoptosis, and we purified the molecule responsible for this effect and determined its chemical structure. Using an MTT assay we showed a rapid (4 h) decrease in the number of viable cells. This cytotoxic effect was due to apoptosis, according to the fragmentation pattern of DNA, Hoechst 33342 staining and FACS analysis of the phosphatidylserine externalization. This apoptosis was blocked by using the caspase inhibitor Z VAD.fmk, indicating the involvement of caspases. Prodigiosin is a red pigment produced by various bacteria including S. marcescens. Using mutants of S. marcescens (OF, WF and 933) that do not synthesize prodigiosin, we further showed that prodigiosin is involved in this apoptosis. This evidence was corroborated by spectroscopic analysis of prodigiosin isolated from S. marcescens. These results indicate that prodigiosin, an immunosuppressor, induces apoptosis in haematopoietic cancer cells with no marked toxicity in nonmalignant cells, raising the possibility of its therapeutic use as an antineoplastic drug. PMID- 11015312 TI - The effect of Gi-protein inactivation on basal, and beta(1)- and beta(2)AR stimulated contraction of myocytes from transgenic mice overexpressing the beta(2)-adrenoceptor. AB - The atria and ventricles of transgenic mice (TGbeta(2)) with cardiac overexpression of the human beta(2)-adrenoceptor (beta(2)AR) were initially reported to show maximum contractility in the absence of beta-AR stimulation. However, we have previously observed a different phenotype in these mice, with myocytes showing normal contractility but reduced betaAR responses. We have investigated the roles of cyclic AMP and Gi in basal and betaAR function in these myocytes. ICI 118,551 at inverse agonist concentrations decreased contraction by 32%. However, the cyclic AMP antagonist Rp-cAMPS had no effect on contraction in TGbeta(2) myocytes, indicating that there was no tonic influence of raised cyclic AMP. These findings cannot be explained by the proposed model for inverse agonism, where the activated receptor (R*) raises cyclic AMP levels and so increases contraction in the absence of agonist. After pertussis toxin (PTX) pretreatment to produce inactivation of Gi, the basal contraction in 1 mM Ca(2+) was increased in TGbeta(2) mice (7.82+/-0.47%, n=23) compared to LM mice (3.60+/ 0.59%, n=11) (P<0.001). The contraction amplitude of myocytes to the maximal concentration of isoprenaline was also increased significantly by PTX in TGbeta(2) mice (9.40+/-1.22%, n=8) and was no longer reduced compared to LM mice (8.93+/-1.50%, n=11). Both beta(1)- and beta(2)AR subtypes were affected both by the original desensitization and by the resensitization with PTX. PTX treatment has therefore restored the original phenotype, with high basal contractility and little further effect of isoprenaline. We suggest that both beta-AR desensitization and lack of increased basal contraction in ventricular myocytes from our colony of TGbeta(2) mice were due to increased activity of PTX-sensitive G-proteins. PMID- 11015313 TI - Regulation of the human prostanoid TPalpha and TPbeta receptor isoforms mediated through activation of the EP(1) and IP receptors. AB - The intermolecular cross-regulation mediated by the prostanoid IP-receptor (IP)/EP(1) receptor (EP(1)) agonists PGI(2) and 17 phenyl trinor PGE(2) on TP receptor (TP) signalling within platelets was compared to that which occurs to the individual TPalpha and TPbeta receptors over-expressed in human embryonic kidney (HEK) 293 cells. Ligand mediated TP receptor activation was monitored by analysing mobilization of intracellular calcium ([Ca(2+)](i)) following stimulation with the selective thromboxane (TX) A(2) mimetic U46619. Consistent with previous studies, in platelets, PGI(2) acting through endogenous IP receptors completely inhibited U46619-mediated TP receptor signalling in a protein kinase (PK) A-dependent, PKC-independent manner. In HEK 293 cells, PGI(2), acting through endogenous AH6809 sensitive EP(1) rather than IP receptors, and the selective EP(1) receptor agonist 17 phenyl trinor PGE(2) antagonized U46619-mediated signalling by both TPalpha and TPbeta receptors in a PKC-dependent, PKA-independent manner. The maximum response induced by either ligand was significantly (P<0.005) greater for the TPalpha receptor than the TPbeta receptor, pointing to possible physiologic differences between the TP isoforms, although the potency of each ligand was similar for both TP receptors. TP(Delta328), a truncated variant of TP receptor lacking the C-tail sequences unique to TPalpha or TPbeta receptors, was not sensitive to EP(1) receptor mediated regulation by PGI(2) or 17 phenyl trinor PGE(2) In conclusion, these data confirm that TPalpha and TPbeta receptors are subject to cross regulation by EP(1) receptor signalling in HEK 293 cells mediated by PKC at sites unique to the individual TP receptors and that TPalpha receptor responses are significantly more reduced by EP(1) receptor regulation than those of the TPbeta receptor. PMID- 11015314 TI - Effects of (+)-pentazocine and 1,3-di-o-tolylguanidine (DTG), sigma (sigma) ligands, on micturition in anaesthetized rats. AB - The effects of two sigma (sigma) binding site ligands, (+)-pentazocine and 1,3-di o-tolylguanidine (DTG), on bladder functions were examined in rats. Cystometry using urethane-anaesthetized rats showed that (+)-pentazocine (1 - 5 mg kg(-1), i.v.) and DTG (1 - 5 mg kg(-1), i.v.) prolonged micturition intervals, indicating increased bladder capacity and raised the threshold pressure. The effects of (+) pentazocine (2 mg kg(-1), i.v. ) on micturition were not influenced by naloxone (0.5 mg kg(-1), i.v. ), which antagonized similar effects of morphine (2 mg kg( 1), i.v.). When administered intracerebroventricularly (i.c.v.), DTG (1 microg) and (+)-pentazocine (30 microg) prolonged micturition intervals with increased threshold pressure on the cystometrogram. In isolated bladder detrusor strips of rats, (+)-pentazocine (3 microM) and DTG (1 microM) did not affect contractile responses to electrical field stimulation. A higher concentration of DTG (3 microM) slightly suppressed the response induced by 30 Hz stimulation. The effects of (+)-pentazocine and DTG on micturition were abolished by pre-treatment with pertussis toxin (PTX, 1 microg, i.c.v.). These results indicate that typical sigma ligands, such as (+)-pentazocine and DTG, increase bladder capacity in anaesthetized rats. Moreover, the mechanism by which sigma ligands change the urinary storage properties in rats may involve pathways in which the function of Gi/o proteins is necessary. PMID- 11015315 TI - The effect of nucleotides and adenosine on stimulus-evoked glutamate release from rat brain cortical slices. AB - Evidence has previously been presented that P1 receptors for adenosine, and P2 receptors for nucleotides such as ATP, regulate stimulus-evoked release of biogenic amines from nerve terminals in the brain. Here we investigated whether adenosine and nucleotides exert presynaptic control over depolarisation-elicited glutamate release. Slices of rat brain cortex were perfused and stimulated with pulses of 46 mM K(+) in the presence of the glutamate uptake inhibitor L-trans pyrrolidine-2,4-dicarboxylic acid (0.2 mM). High K(+) substantially increased efflux of glutamate from the slices. Basal glutamate release was unchanged by the presence of nucleotides or adenosine at concentrations of 300 microM. Adenosine, ATP, ADP and adenosine 5'-O-(3-thiotriphoshate) at 300 microM attenuated depolarisation-evoked release of glutamate. However UTP, 2-methylthio ATP, 2 methylthio ADP, and alpha,beta-methylene ATP at 300 microM had no effect on stimulated glutamate efflux. Adenosine deaminase blocked the effect of adenosine, but left the response to ATP unchanged. The A(1) antagonist 8-cyclopentyl-1, 3 dipropylxanthine antagonised the inhibitory effect of both adenosine and ATP. Cibacron blue 3GA inhibited stimulus-evoked glutamate release when applied alone. When cibacron blue 3GA was present with ATP, stimulus-evoked glutamate release was almost eliminated. However, this P2 antagonist had no effect on the inhibition by adenosine. These results show that the release of glutamate from depolarised nerve terminals of the rat cerebral cortex is inhibited by adenosine and ATP. ATP appears to act directly and not through conversion to adenosine. PMID- 11015316 TI - Vagal neurotransmission to the ferret lower oesophageal sphincter: inhibition via GABA(B) receptors. AB - GABA(B) receptors modulate the function of the lower oesophageal sphincter (LOS) in vivo by inhibiting neurotransmitter release in the vagal pathway controlling LOS relaxation. We aimed to determine whether this effect was mediated peripherally on vagal motor outflow to the ferret LOS in vitro. The LOS, with intact vagal innervation, was prepared from adult ferrets and LOS tension measured. Vagal stimulation (0.5 - 10 Hz, 30 V) evoked a tetrodotoxin-sensitive, frequency-dependent relaxation. Both GABA (3x10(-4) M) and (+/-)baclofen (2x10( 4) M) inhibited vagally-stimulated LOS relaxation. The potent GABA(B) receptor selective agonist 3-APPA dose-dependently inhibited vagally-stimulated LOS relaxation, with an EC(50) value of 0.7 microM Decreased responses following vagal stimulation in the presence of (+/-)baclofen or 3-APPA were reversed with the potent GABA(B) receptor antagonist CGP 62349. Neither CGP 62349 nor muscimol (GABA(A) receptor agonist) alone affected LOS responses following vagal stimulation. Agonists of other G protein-coupled receptors (clonidine (alpha(2) adrenoceptor) (5x10(-6) M), U50488 (kappa opioid) (10(-5) M), neuropeptide Y (10( 6) M)) did not affect vagally-mediated LOS relaxation. The present study supports a discrete presynaptic inhibitory role for GABA(B) receptors on vagal preganglionic fibres serving inhibitory motorneurones in the ferret LOS. PMID- 11015317 TI - Suppression of the development of hypertension by the inhibitor of inducible nitric oxide synthase. AB - Our previous study demonstrated that the aortic inducible nitric oxide synthase (iNOS) expression and the plasma nitrite level in spontaneously hypertensive rats (SHR) were greater than that in age-matched Wistar-Kyoto rats (WKY). We subsequently hypothesized that the over-expression of iNOS might play an important role in the pathogenesis of hypertension in SHR. In the present study, pyrrolidinedithiocarbamate (PDTC, 10 mg kg(-1) day(-1), p.o., antioxidant and nuclear factor-kappa B inhibitor) and aminoguanidine (15 mg kg(-1) day(-1), p.o., selective inhibitor of iNOS) was used to treat SHR and WKY from age of 5 weeks through 16 weeks. We found that PDTC and aminoguanidine significantly suppressed the development of hypertension and improved the diminished vascular responses to acetylcholine in SHR but not in WKY. Likewise, the increase of iNOS expression, nitrotyrosine immunostaining, nitric oxide production and superoxide anion formation in adult SHR were also significantly suppressed by chronic treatment with PDTC and aminoguanidine. In conclusion, this study demonstrated that both PDTC and aminoguanidine significantly attenuated the development of hypertension in SHR. The results suggest that PDTC suppresses iNOS expression due to its anti oxidant and/or nuclear factor-kappa B inhibitory properties. However, the effect of aminoguanidine was predominantly mediated by inhibition of iNOS activity, thereby reducing peroxynitrite formation. We propose that the development of a more specific and potent inhibitor of iNOS might be beneficial in preventing pathological conditions such as the essential hypertension. PMID- 11015318 TI - Pharmacological analysis by HOE642 and KB-R9032 of the role of Na(+)/H(+) exchange in the endothelin-1-induced Ca(2+) signalling in rabbit ventricular myocytes. AB - The role of Na(+)/H(+) exchange in endothelin-1 (ET-1)-induced increases in Ca(2+) transients and cell shortening was studied in rabbit ventricular myocytes loaded with indo-1/AM. Selective inhibitors of Na(+)/H(+) exchange HOE642 (4 isopropyl-3-methyl-sulphonylbenzoyl guanidine methanesulphonate) and KB-R9032 (N (4-isopropyl-2,2-dimethyl-3-oxo-3, 4-dihydro-2H-benzo-[1,4]oxazine-6-carbonyl) guanidine methanesulphonate) were used as pharmacological tools for the analysis. ET-1 at 0.1 nM induced an increase in Ca(2+) transients by 45.6%, while it increased cell shortening by 109.6%. For a given increase in cell shortening, the ET-1-induced increase in Ca(2+) transients was much smaller than that induced by isoprenaline (ISO, 10 nM). Pretreatment with HOE642 and KB-R9032 (1 microM) inhibited the increase in cell shortening induced by 0.1 nM ET-1 by 51 and 65. 4%, respectively, without a significant alteration of ET-1-induced increase in Ca(2+) transients. HOE642 and KB-R9032 did not affect baseline levels of cell shortening and peak Ca(2+) transients, and the effects of ISO (10 nM). These results indicate that activation of Na(+)/H(+) exchange by ET-1 may play an important role in the positive inotropic effect and the ET-1-induced increase in myofilament Ca(2+) sensitivity in rabbit ventricular myocytes. PMID- 11015319 TI - Cannabinoid receptor-mediated inhibition of acetylcholine release from hippocampal and cortical synaptosomes. AB - In previous studies cannabinoid agonists have been found to inhibit and cannabinoid antagonists to enhance electrically-evoked [(3)H]-acetycholine (ACh) release in hippocampal slices. The present study was undertaken to determine if similar cannabinoid effects could be observed in synaptosomes. [(3)H]-ACh release was evoked by two methods, both sensitive to presynaptic receptor effects. The first involved the addition of 1.3 mM calcium following perfusion with calcium free Krebs and the second the addition of 11 mM potassium following perfusion with normal Krebs. In hippocampal synaptosomes the 1.3 mM calcium-evoked release and the high potassium-evoked [(3)H]-ACh release were inhibited by the cannabinoid agonist, WIN 55212-2, by 59 and 39%, respectively, and with an EC(50) of approximately 1 nM. WIN 55212-2 produced a similar, although less potent, inhibition of [(3)H]-ACh release in cortical synaptosomes. No inhibitory effect of WIN 55212-2 on [(3)H]-ACh release in striatal synaptosomes was observed, supporting previous data collected in this area with brain slices. The cannabinoid antagonist, SR 141716A, produced a robust enhancement of 1.3 mM calcium-evoked [(3)H]-ACh release in hippocampal synaptosomes (EC(50)<0.3 nM) but had no effect on potassium-evoked release or on [(3)H]-ACh release in the cortex or striatum. In conclusion our data demonstrates the inhibitory effects of WIN 55212-2 observed on ACh release in brain slices can be observed in hippocampal and cortex synaptosomes, suggesting a direct action of these compounds on the synaptic terminals. The SR 141716A-induced enhancement of ACh release can similarly be observed in hippocampal synaptosomes and is probably due to an inverse agonist action at constitutively active receptors. PMID- 11015320 TI - Psychometrics of odor quality discrimination: method for threshold determination. AB - There is no natural physical continuum for odor quality along which an odor quality discrimination (OQD) threshold can be measured. In an attempt to overcome this problem, the substitution-reciprocity (SURE) method defines a framework for the measurement of an OQD threshold. More specifically, it (i) defines a threshold concept for OQD, including the quantification of qualitative change of the stimulus, and (ii) suggests how to avoid perceived intensity as an unwanted cue for discrimination. In doing this, the psychometric properties of odor quality in the case of eugenol and citral are investigated using both discrimination (experiment 1) and scaling (experiment 2). Based on discriminatory responses, a change of approximately one-third in stimulus content was needed to reach the OQD threshold for eugenol and citral. PMID- 11015321 TI - Glossopharyngeal taste responses of the channel catfish to binary mixtures of amino acids. AB - This study examines the neural processing of binary mixtures in the glossopharyngeal (IX) taste system of the channel catfish, Ictalurus punctatus, and finds that the nature of the components of a mixture determines the intensity of the neural response to it. Taste buds in fish innervated by IX are located along the gill rakers of the first gill arch and rostral floor of the oral cavity, and function primarily in the consummatory phase of feeding behavior; however, few studies of IX taste responses have been reported in any species of teleost. Here, we report IX taste responses to eight different binary mixtures of amino acids whose components were adjusted to be approximately equipotent in electrophysiological recordings. Four binary (group I) mixtures whose components were indicated from prior electrophysiological cross-adaptation experiments to bind to independent receptor sites resulted in significantly larger (22% average increase) integrated IX taste activity than four other (group II) binary mixtures whose components were indicated to bind to the same or highly cross-reactive receptor sites. These results are similar to those observed previously from facial nerve recordings in channel catfish, and to olfactory and taste responses in other vertebrate and invertebrate species. The group I results help to explain behavioral observations that chemical mixtures of chemosensory stimuli are often more stimulatory than their individual components. PMID- 11015322 TI - Taste preference synergy between glutamate receptor agonists and inosine monophosphate in rats. AB - Monosodium glutamate (MSG) elicits a taste called umami and interacts synergistically with nucleotide monophosphates such as 5'-inosine monophosphate (IMP) to potentiate this taste intensity. Indeed, the synergistic interaction of nucleotide monophosphates and MSG is a hallmark of umami. We examined interactions between MSG and other taste stimuli, including IMP, by measuring the lick rates of non-deprived rats during 30 s trials. To control for non-linear psychophysical functions, the concentration of one taste stimulus in a binary mixture was systematically increased while the concentration of the second taste stimulus was decreased (stimulus substitution method). Synergy between two stimuli was detected if the lick rate for a binary mixture exceeded that expected from the sum of the lick rates for each stimulus alone. In initial experiments, taste synergy was observed when rats were presented with mixtures of MSG and IMP but not with mixtures of MSG and sucrose. In subsequent experiments, glutamate receptor agonists other than MSG were presented with IMP to test for taste synergy. No evidence of synergy was seen when rats were presented with mixtures of IMP and kainic acid or IMP and N:-methyl-D-aspartate. However, taste synergy between IMP and L-AP4, a potent agonist at mGluR4 receptors, was observed. These results suggest that a metabotropic glutamate receptor similar to mGluR4 may be involved in the taste synergy that characterizes umami. PMID- 11015323 TI - Right-nostril dominance in discrimination of unfamiliar, but not familiar, odours. AB - In a recent PET study on processing of unfamiliar odours we observed that odour discrimination performance was superior during right compared with left nostril presentations, and that mainly the right cerebral hemisphere was activated. In the present study we investigated whether the asymmetric performance is present also during the processing of familiar odours. Seventy-one right-handed healthy subjects (age 21-49 years, 40 females) with normal nasal anatomy and olfactory thresholds participated. Forty pairs of odours (20 familiar and 20 unfamiliar) were presented in the same/different paradigm, alternating nostrils and balancing the order. The number of errors during the discrimination task was compared with respect to nostril and odour familiarity. The overall odour discrimination performance was superior on the right side. However, this difference was valid only for unfamiliar odours, whereas the performance for familiar odours was symmetrical. Familiar odours were easier to discriminate than unfamiliar ones. The present data are congruent with the idea of a semantic influence on odour processing. Odours seem to be processed with a right sided preponderance when not clearly familiar, and symmetrically when language becomes involved. Future studies on odour processing should therefore take into account odour familiarity and side of presentation. PMID- 11015324 TI - Comprehensive study on G protein alpha-subunits in taste bud cells, with special reference to the occurrence of Galphai2 as a major Galpha species. AB - Previous studies have identified many cDNA species that encode a variety of G protein alpha-subunits occurring in taste buds. These include the cDNA encoding a taste-bud-specific Galpha, gustducin (G(gust)). Here we carried out comprehensive analyses of Galpha species that occur in the taste buds of rat circumvallate papillae and also in their single cells isolated from the taste buds. Reverse transcriptase-polymerase chain reaction showed the presence of 10 kinds of Galpha cDNAs, including a splice variant of Galphas, among which G(gust), Galphas, Galphai2 and Galphai3 cDNAs were shown to be major species. In situ hybridization and immunohistochemistry showed that Galphai2, as well as G(gust), expressed in a subset of taste bud cells, and the frequency of Galphai2-expression appears to be higher than that of G(gust). Southern analyses of the amplified cDNA from single cells showed that each taste bud cell expresses multiple Galpha mRNA species. For example, some Galphai2-positive cells also express one or more other Galpha species, including Galphas, Galphai3 and G(gust), and there is no apparent correlation in expression among the three Galpha species. PMID- 11015325 TI - Amino acids dissolved in stream water as possible home stream odorants for masu salmon. AB - It is well established that salmon return to their home stream by sensing the odors of the stream water. In this study we have attempted to identify the home stream odorants used by masu salmon in Lake Toya. The salmon in Lake Toya return to the home stream which flows into the lake after lake life for 2-3 years. Besides water from the home stream, waters from two other streams which flow into Lake Toya were also used in the experiments. We analyzed the compositions of amino acids, inorganic cations and bile acids in waters from the three streams. Application of mixtures of inorganic cations or bile acids, reconstituted based on the compositions of the stream waters, to the olfactory epithelium induced only very small responses. On the other hand, application of mixtures of amino acids induced large responses. The response to artificial stream water reconstituted based on the compositions of amino acids and salts closely resembles that to the corresponding stream water. Cross-adaptation experiments with three combinations of the mixtures were carried out. The response pattern for each combination closely resembled that to the corresponding combination of stream waters. Based on the results obtained, we concluded that amino acids dissolved in the home stream water are possible home stream odorants. PMID- 11015326 TI - Chemosensory proteins from the proboscis of mamestra brassicae. AB - Soluble, low molecular weight proteins were immunodetected in proboscis extracts of Mamestra brassicae males by Western blot, using antibodies raised against the general odorant-binding protein of the moth Antheraea polyphemus. The same antibodies weakly labelled the sensillum lymph and subcuticular space of sensilla styloconica on ultrathin sections of the proboscis. The morphology of sensilla styloconica is described. The immunodetected proteins yielded several N-terminal sequences, three of which showed strong affinity for tritiated analogues of pheromonal compounds of M. brassicae in binding assays. The cDNAs coding for these sequences were cloned and it was shown that the new proteins are related to the OS-D protein of Drosophila. They are named chemosensory proteins (CSP-MBRA:A1 CSP-MBRA:A5 and CSP-MBRA:B1 and CSP-MBRA:B2) and may have an odorant-binding protein-like function. A common localization in both olfaction and taste organs suggests a physiological role depending on the cellular environment. PMID- 11015327 TI - Neohesperidin dihydrochalcone is not a taste enhancer in aqueous sucrose solutions. AB - Neohesperidin dihydrochalcone (NHDC) is an intensive sweetener, obtained by alkaline hydrogenation of neohesperidin. In this investigation a supposed taste enhancing effect of this substance was tested. A three-step procedure was used. In the first experiment, using a pool of 31 subjects, NHDC and sucrose detection thresholds were measured. In the second experiment, psychophysical functions for both tastants were determined. Then, 15 participants closest to the group threshold who, in addition, had produced monotonic psychophysical taste functions were selected to participate in the next two experiments. In the third experiment, taste enhancement was tested. Three psychophysical sucrose functions were constructed, one with a near-threshold amount of NHDC added to each of seven sucrose concentrations, one with a near-threshold amount of sucrose added (control 1) and one without any addition (control 2). No difference was found between the NHDC-enriched sucrose function and the sucrose-enriched sucrose function. Finally, in experiment 4, differential threshold functions were constructed with either NHDC or sucrose added. Neither the overall shape of the functions nor a comparison of the points of subjective equality showed enhancement. It was concluded that weak NHDC does not enhance the taste of aqueous sucrose solutions. PMID- 11015328 TI - Exposure to behaviourally relevant odour reveals differential characteristics in rat central olfactory pathways as studied through oscillatory activities. AB - This study investigated how changes in nutritional motivation modulate odour related oscillatory activities at several levels of the olfactory pathway in non trained rats. Local field potential recordings were obtained in freely moving animals in the olfactory bulb (OB), anterior and posterior parts of the piriform cortex (APC and PPC respectively) and lateral entorhinal cortex (EC). Dynamic signal analysis detected changes in power during odour presentation for several frequency bands The results showed that in most cases odour presentation was associated with changes in a wide 15-90 Hz frequency band of activity in each olfactory structure. However, nutritional state modulated initial responses to food odour (FO) in the OB and EC selectively in the 15-30 Hz frequency band. Changes in nutritional state also modulated responses to repeated FO stimuli. Habituation was expressed differentially across structures with a clear dissociation between the two parts of the piriform cortex. Finally, systemic injections of scopolamine (0.125 mg/kg) selectively blocked expression of the nutritional modulation in the OB found in the beta band. These results suggest that internal state can differentially modulate odour processing among different olfactory areas and point to a cholinergic-sensitive beta band oscillation during presentation of a behaviourally meaningful odorant. PMID- 11015329 TI - Taste function in patients with oral burning. AB - Burning mouth syndrome (BMS) is an oral pain disorder occurring primarily in post menopausal women and is frequently accompanied by taste complaints. This association of symptoms suggests an interaction between the mechanisms of nociception and gustation, two senses with strong hedonic components. Seventy three patients of the Taste and Smell Clinic at the University of Connecticut Health Center who reported experiencing 'unexplained oral burning' were evaluated for taste function. Both intensity ratings and quality identifications were measured for a concentration series of sucrose ('sweet'), NaCl ('salty'), citric acid ('sour') and quinine-HCl ('bitter'). The 57 women with BMS gave lower intensity ratings to NaCl and sucrose than comparably aged, same sex controls. Concentrations of NaCl and sucrose >0.10 M were most affected; concentrations of sucrose and NaCl <0.10 M were rated similarly by BMS and control women. No intensity differences were found for citric acid or quinine-HCl at any concentration and no differences were evident between the 16 BMS men and the 14 control men for any stimulus. The BMS women also misidentified the quality of 19% of the stimuli that were detected whereas control women misidentified 8%. Both groups detected a similar proportion of stimuli and found lower stimulus concentrations more difficult to identify than higher concentrations. Identification of NaCl as 'salty' and citric acid as 'sour' was particularly difficult for BMS women. The present findings are consistent with the hypothesis that pain pathway activation may affect neural and behavioral taste function. PMID- 11015330 TI - The origin of slow potentials on the tongue surface induced by frog glossopharyngeal efferent fiber stimulation. AB - When the glossopharyngeal (GP) nerve of the frog was stimulated electrically, electropositive slow potentials were recorded from the tongue surface and depolarizing slow potentials from taste cells in the fungiform papillae. The amplitude of the slow potentials was stimulus strength- and the frequency dependent. Generation of the slow potentials was not related to antidromic activity of myelinated afferent fibers in the GP nerve, but to orthodromic activity of autonomic post-ganglionic C fibers in the GP nerve. Intravenous injection of atropine abolished the positive and depolarizing slow potentials evoked by GP nerve stimulation, suggesting that the slow potentials were induced by the activity of parasympathetic post-ganglionic fibers. The amplitude and polarity of the slow potentials depended on the concentration of adapting NaCl solutions applied to the tongue surface. These results suggest that the slow potentials recorded from the tongue surface and taste cells are due to the liquid junction potential generated between saliva secreted from the lingual glands by GP nerve stimulation and the adapting solution on the tongue surface. PMID- 11015331 TI - Twenty-second annual meeting of the association for chemoreception sciences PMID- 11015333 TI - Myocardial ischemia and lipoprotein lipase activity. PMID- 11015334 TI - Fetoscopic direct fetal cardiac access in sheep : An important experimental milestone along the route to human fetal cardiac intervention. AB - BACKGROUND: Fetal cardiac interventions by direct ultrasound-guided approaches or open fetal cardiac surgery have been fraught with technical difficulties, as well as with significant maternal and fetal morbidity in humans. Therefore, the purpose of our study in sheep was to assess the feasibility and potential of fetoscopic direct fetal cardiac access. METHODS AND RESULTS: In 15 anesthetized pregnant ewes (88 to 109 days of gestation; term, 145 days), 3 to 4 trocars were percutaneously placed in the uterus. Using videofetoscopic equipment, we assessed the feasibility of achieving direct fetal cardiac access. Minimally invasive direct fetal cardiac access by operative fetoscopy was achieved in 10 of the 15 fetal sheep. In 7 fetuses, the approach was successfully tested for fetal cardiac pacing (n=5) or antegrade fetal cardiac catheterization (n=2). Access was not achieved in 5 fetuses because of bleeding complications (n=2) or because the fetoscopic setup could not be established (n=3). All but 2 fetal sheep were alive at the end of the procedure. Acute fetal demise resulted from maternal hypotension or kinking of the fetal inferior caval vein by sternal suspension. Six ewes continued gestation; 3 of these went to term, with a normal fetal outcome. Two ewes died from septicemia 3 and 7 days after the procedure, and 1 ewe aborted 1 month after the procedure. CONCLUSIONS: Minimally invasive direct fetal cardiac access by operative fetoscopy is feasible in fetal sheep. The fetoscopic approach carries important potential for fetal cardiac pacing, antegrade fetal valvuloplasties, and resection of fetal intrapericardial teratomas in human fetuses. PMID- 11015335 TI - Therapeutic angiogenesis with recombinant fibroblast growth factor-2 improves stress and rest myocardial perfusion abnormalities in patients with severe symptomatic chronic coronary artery disease. AB - BACKGROUND: We report the effects of the administration of recombinant fibroblast growth factor-2 (rFGF-2) protein on myocardial perfusion using single photon emission computed tomography imaging in humans with advanced coronary disease. METHODS AND RESULTS: A total of 59 patients with coronary disease that was not amenable to mechanical revascularization underwent intracoronary (n=45) or intravenous (n=14) administration of rFGF-2 in ascending doses. Changes in perfusion were evaluated at baseline and again at 29, 57, and 180 days after rFGF 2 administration. In this uncontrolled study, perfusion scans were analyzed by 2 observers who were blinded to patient identity and test sequence; scans were displayed in random order, with scans from nonstudy patients randomly interspersed to enhance blinding. Combining all dose groups, a reduction occurred in the per-segment reversibility score (reflecting the magnitude of inducible ischemia) from 1.7+/-0.4 at baseline to 1.1+/-0.6 at day 29 (P:<0.001), 1.2+/-0.7 at day 57 (P:<0.001), and 1.1+/-0.7 at day 180 (P:<0.001). The 37 patients with evidence of resting hypoperfusion had evidence of improved resting perfusion: their per-segment rest perfusion score of 1.5+/-0. 5 at baseline decreased to 1.0+/-0.8 at day 29 (P:<0.001), 1.0+/-0.8 at day 57 (P:=0.003), and 1.1+/-0.9 at day 180 (P:=0.11). CONCLUSIONS: These preliminary data suggest that the administration of rFGF-2 to patients with advanced coronary disease resulted in an attenuation of stress-induced ischemia and an improvement in resting myocardial perfusion; these findings are consistent with a favorable effect of therapeutic angiogenesis. PMID- 11015336 TI - Elevated troponin I level on admission is associated with adverse outcome of primary angioplasty in acute myocardial infarction. AB - BACKGROUND: In patients with acute myocardial infarction (AMI) undergoing thrombolytic therapy, an elevated troponin level on admission is associated with a lower reperfusion rate and a complicated clinical course. Whether an elevated troponin level on admission similarly predicts an adverse outcome in patients undergoing primary angioplasty is currently unknown and was investigated in the present study. METHODS AND RESULTS: Cardiac troponin I (cTnI) was determined on admission in 110 consecutive patients with AMI associated with ST-segment elevation or left bundle branch block who underwent primary angioplasty. Fifty four patients (49%) had an elevated cTnI (>/=0.4 ng/mL) on admission. In patients with elevated cTnI, primary angioplasty was less likely to achieve TIMI 3 flow (as classified by the Thrombolysis in Myocardial Infarction trial) in univariate (76% versus 96%, P:=0.03) or in multivariate (odds ratio 0.1, 95% CI 0.02 to 0.54) analysis. Patients with elevated cTnI were more likely to develop congestive heart failure (23% versus 9%, P:<0.05) and death, heart failure, or shock (30% versus 9%, P:=0.006). Elevated cTnI remained a significant predictor of the composite end point after controlling for other clinical data that were available early in the course, including time to presentation and angiographic results (relative risk 5.2, 95% CI 1.03 to 26.3). During a follow-up of 426+/-50 days, elevated admission cTnI was a predictor of cardiac mortality (11% versus 0%, P:=0.012), adverse cardiac events (cardiac mortality or nonfatal reinfarction; 19% versus 5.4%, P:=0.04), and adverse cardiac events plus target vessel revascularization (32% versus 14%, P:=0.054). CONCLUSIONS: In patients with ST-segment elevation AMI, an elevated cTnI on admission is associated with an increased risk of primary angioplasty failure and a more complicated clinical course. PMID- 11015338 TI - Cardiorespiratory fitness and coronary heart disease risk factors: the LDS Hospital Fitness Institute cohort. AB - BACKGROUND: Cardiorespiratory fitness is favorably associated with most modifiable coronary heart disease (CHD) risk factors. Findings are limited, however, by few data for women, persons with existing CHD, and low-risk populations. In the present study, we described cross-sectional associations between cardiorespiratory fitness and CHD risk factors in a large cohort of middle-aged men and women, of whom the majority were LDS Church members (Mormons), with and without existing CHD. METHODS AND RESULTS: Comprehensive health examinations were performed on 3232 men (age 45.9+/-10.8 years) and 1128 women (age 43.8+/-12.8 years) between 1975 and 1997. Maximal treadmill exercise testing was used to categorize those with (12% of the men and 10% of the women) and those without CHD into age- and sex-specific cardiorespiratory fitness quintiles. After adjustments for age, body fat, smoking status, and family history of CHD, favorable associations were observed between fitness and most CHD risk factors among men and women, regardless of CHD status. CONCLUSIONS: These data indicate that enhanced levels of cardiorespiratory fitness may confer resistance to elevations in CHD risk factors even in a low-risk sample of middle aged men and women. Furthermore, these findings reinforce current public health recommendations that advocate increased national levels of physical activity and cardiorespiratory fitness for primary and secondary CHD prevention. PMID- 11015337 TI - 17beta-estradiol decreases endothelin-1 levels in the coronary circulation of postmenopausal women with coronary artery disease. AB - BACKGROUND: Estrogen reverses acetylcholine-induced coronary vasoconstriction via the possible facilitation of endothelium-derived NO. Estrogen also affects endothelium-derived constrictor factors. We therefore investigated the effects of 17beta-estradiol on coronary vasomotor responses to substance P (SP), and coronary sinus endothelin-1 and NO metabolite levels in postmenopausal women with coronary heart disease. METHODS AND RESULTS: We studied 20 women; 14 received estrogen (mean age 65+/-2 years) and 6 served as ethanol control subjects (age 63+/-3 years). Intracoronary infusions of papaverine (8 mg) and SP were administered before and 20 minutes after 50 pg/min 17beta-estradiol or 0.2 microL/min control. Coronary blood flow was calculated from the diameter, as measured with quantitative coronary angiography, and flow velocity, as measured with intracoronary Doppler. Coronary sinus plasma endothelin-1 and nitrite/nitrate (NO(2)/NO(3)) were measured at baseline, at peak velocity response to each infusion, and every 5 minutes during the estradiol infusion. Endothelin-1 levels were decreased after 20 minutes of estradiol (1.12+/-0.18 versus 0.86+/-0.17 pmol/L baseline2 versus estradiol, P:=0.05). Endothelin-1 levels to SP decreased after 17beta-estradiol (1.29+/-0. 18 versus 1.04+/-0.15 and 1.3+/-0.16 versus 0.99+/-0.17 pmol/L for before versus after estradiol, 10 and 25 pmol/min SP; both P:<0.05). NO(2)/NO(3) levels did not change. There was no change in vasomotor responses to estradiol alone or to papaverine or SP before versus after estradiol. CONCLUSIONS: Short-term intracoronary 17beta-estradiol administration decreases coronary endothelin-1 levels. There was no enhancement of vasomotor responses to SP after the administration of estrogen, suggesting that the effects of estrogen on coronary acetylcholine responses may be a specific and not a generalized effect on coronary vasoreactivity. PMID- 11015339 TI - Lipoprotein lipase activity is associated with severity of angina pectoris. REGRESS Study Group. AB - BACKGROUND: Raised triglyceride-rich lipoproteins significantly increase the risk for cardiovascular disease. Variation in the activity of the enzyme lipoprotein lipase (LPL), which is crucial in the removal of these lipoproteins, may therefore modulate this risk. METHODS AND RESULTS: Postheparin levels of LPL activity and mass were measured in a large cohort of male coronary artery disease patients participating in the Regression Growth Evaluation Statin Study (REGRESS), a lipid-lowering regression trial. In addition, the relationships between LPL activity and mass and severity of angina pectoris according to the NYHA classification and silent ischemia on 24-hour ambulatory ECG monitoring were assessed. Patients in different LPL activity quartiles and mass had different severities of angina; a total of 47% of patients in the lowest LPL quartile reported class III or IV angina. In contrast, only 29% in the highest activity quartile (P:=0.002) had severe angina. These parameters were supported by ambulatory ECG results, for which the total ischemic burden in the lowest LPL activity quartile was 36. 5+/-104.1 mm x min compared with 14.8+/-38.8 mm x min in the highest quartile of LPL activity (P:=0.001). LPL activity levels were strongly correlated with LPL mass (r=0.70, P:<0.0001). A significant association between the LPL protein mass and NYHA class (P:=0.012) was also demonstrated. CONCLUSIONS: We have demonstrated a significant relationship between LPL mass and activity and severity of ischemia as defined by angina class and ambulatory ECG. These results suggest that LPL influences risk for coronary artery disease by both catalytic and noncatalytic mechanisms. PMID- 11015341 TI - Development of antibody against epitope of lipoprotein(a) modified by oxidation: evaluation of new enzyme-linked immunosorbent assay for oxidized lipoprotein(a). AB - BACKGROUND: Recently, the biological effects of oxidized lipoprotein(a) [Lp(a)] have been reported to be more potent than Lp(a), the arteriosclerosis-relevant lipoprotein. Thus, investigations with oxidized Lp(a) are expected to provide viewpoints different from the conventional ones based on Lp(a). METHODS AND RESULTS: An anti-Lp(a) monoclonal antibody (161E2) was produced against synthetic peptide antigen (Arg-Asn-Pro-Asp-Val-Ala-Pro). This epitope was characterized as having various properties because its external exposure was induced as a result of oxidative modification. Using 161E2 antibody, we developed a new enzyme-linked immunosorbent assay to measure Lp(a) modified by oxidative stress. The present data demonstrated that oxidized Lp(a) that contains the epitope of 161E2 antibody was present in the serum of humans. Therefore, we used this new enzyme-linked immunosorbent assay to evaluate the role of oxidized Lp(a) in patients with hypertension, which induces oxidative stress. Interestingly, hypertensive patients with complications showed a significantly higher level of oxidized Lp(a) in serum than did normotensive subjects (P:<0.01), whereas there was no significant difference in native Lp(a) between normotensive and hypertensive subjects. Importantly, positive immunostaining with 161E2 monoclonal antibody was found in the human arteriosclerotic tissue. CONCLUSIONS: We developed a new antibody against an epitope in Lp(a) as a result of oxidation treatment but not in native Lp(a). The present data demonstrated in vivo the presence of oxidized Lp(a) in the atherosclerotic tissue and its elevation in hypertensive patients. The presence of oxidized Lp(a) may be important in understanding the role of Lp(a) in cardiovascular disease. PMID- 11015340 TI - Association of fibrinogen with cardiovascular risk factors and cardiovascular disease in the Framingham Offspring Population. AB - BACKGROUND: Fibrinogen has been identified as an independent risk factor for cardiovascular disease and associated with traditional cardiovascular risk factors. Also, the role of elevated fibrinogen in thrombosis suggests that it may be on the causal pathway for certain risk factors to exert their effect. These associations remain incompletely characterized. Moreover, the optimal fibrinogen assay for risk stratification is uncertain. METHODS AND RESULTS: In 2632 subjects from cycle 5 of the Framingham Offspring Population, fibrinogen levels were determined with a newly developed immunoprecipitation test (American Biogenetic Sciences) and the functional Clauss method. With the immunoprecipitation method, there were significant linear trends across fibrinogen tertiles (P:<0.001) for age, body mass index, smoking, diabetes mellitus, total cholesterol, HDL cholesterol, and triglycerides in men and women. The Clauss method had significant results (P:<0.030), except for triglycerides in men. Fibrinogen levels were higher for those with compared with those without cardiovascular disease. After covariate adjustment, fibrinogen remained significantly higher in those with cardiovascular disease with the use of the immunoprecipitation test (P:=0.035 and P:=0.018 for men and women, respectively) but not with the Clauss method. CONCLUSIONS: Fibrinogen was associated with traditional cardiovascular risk factors. Elevation of fibrinogen may provide a mechanism for risk factors to exert their effect. Also, fibrinogen levels were higher among subjects with cardiovascular disease compared with those without disease. The immunoprecipitation test showed a stronger association with cardiovascular disease than the Clauss method, suggesting that it may be a useful screening tool to identify individuals at increased thrombotic risk. PMID- 11015342 TI - Homozygosity for 807 T polymorphism in alpha(2) subunit of platelet alpha(2)beta(1) is associated with increased risk of cardiovascular mortality in high-risk women. AB - BACKGROUND: Platelet adhesion to collagen is the initial step in both hemostasis and thrombosis; this adhesion is mediated by alpha(2)beta(1) on the surface of platelet membranes. An 807 C to T single nucleotide exchange polymorphism close to the gene coding for the alpha(2) subunit of alpha(2)beta(1) is associated with the density of alpha(2)beta(1) on the platelet membrane. METHODS AND RESULTS: We studied the relation of the alpha(2)beta(1) 807 C/T genotype to cardiovascular mortality in a prospective cohort study of 12 239 women who were invited for the breast cancer screening program of Utrecht, the Netherlands. The initial age was between 52 and 67 years. Women were followed on vital status between 1976 and 1995 (168 513 women-years). Data were analyzed by using a nested case-control design. The alpha(2)beta(1) 807 C/T genotype was not associated with cardiovascular mortality in the total population: the rate ratio for cardiovascular mortality in 807 TT homozygotes compared with 807 CC wild types was 1.2 (95% CI 0.8 to 1.7). However, the alpha(2)beta(1) 807 T polymorphism was associated with an increased risk of cardiovascular mortality in women who smoked or in women who had indications of compromised endothelium, such as diabetes and microalbuminuria. In those who were exposed to >/=2 of these factors, the risk ratio (95% CI) between alpha(2)beta(1) 807 TT homozygotes and 807 CC wild types was 14.1 (5.0 to 39.9). CONCLUSIONS: alpha(2)beta(1) 807 TT homozygosity, coding for increased alpha(2)beta(1) density on the platelet membrane, is associated with an increased risk of cardiovascular mortality in those women with indications of compromised endothelium. PMID- 11015343 TI - Time to presentation with acute myocardial infarction in the elderly: associations with race, sex, and socioeconomic characteristics. AB - BACKGROUND: Although prompt treatment is a cornerstone of the management of acute myocardial infarction (AMI), prior studies have shown that one fourth of AMI patients arrive at the hospital >6 hours after symptom onset. It would be valuable to identify individuals at highest risk for late arrival, but predisposing factors have yet to be fully characterized. METHODS AND RESULTS: Data from the Cooperative Cardiovascular Project, involving Medicare beneficiaries aged >65 years hospitalized between January 1994 and February 1996 with confirmed AMI, were used to identify patients who presented "late" (>/=6 hours after symptom onset). Patient characteristics were tested for associations with late presentation by use of backward stepwise logistic regression. Among 102 339 subjects, 29.4% arrived late. Significant predictors of late arrival (odds ratio, 95% CI) included diabetes (1.11, 1.07 to 1.14) and a history of angina (1.32, 1.28 to 1.35), whereas prior MI (0.82, 0.79 to 0.85), prior angioplasty (0.80, 0.75 to 0.85), prior bypass surgery (0.93, 0.89 to 0.98), and cardiac arrest (0.52, 0.46 to 0. 58) predicted early presentation. Additionally, initial evaluation at an outpatient clinic (2.63, 2.51 to 2.75) and daytime presentation (1.67, 1.59 to 1.72) predicted late arrival. Finally, female sex, black race, and poverty, which were evaluated with an 8-level race-sex-socioeconomic status interaction term, were also risk factors for delay. CONCLUSIONS: Delayed hospital presentation is a common problem among Medicare beneficiaries with AMI. Factors associated with delay include not only clinical and logistical issues but also race, sex, and socioeconomic characteristics. Education efforts designed to hasten AMI treatment should be directed at individuals with risk factors for late arrival. PMID- 11015344 TI - Feasibility of a novel blood noise reduction algorithm to enhance reproducibility of ultra-high-frequency intravascular ultrasound images. AB - BACKGROUND: Ultra-high-frequency (40- to 50-MHz) intravascular ultrasound (IVUS) improves image quality compared with conventional 20- to 30-MHz IVUS. However, as the frequency of IVUS increases, high-intensity backscatter from blood components may cause visual difficulties in discrimination between the lumen and arterial wall structure. The purpose of this study was to evaluate the effect of a novel blood noise reduction algorithm (BNR) on quantitative coronary ultrasound measurements. METHODS AND RESULTS: IVUS studies using a 40-MHz transducer were performed in 35 patients with coronary artery disease. A total of 620 gray-scale images (310 pairs) were processed with and without the BNR, and lumen cross sectional area (CSA) was determined by 2 independent observers. With the BNR, the intraobserver and interobserver correlation coefficients for lumen CSA were significantly improved (0.85 to 0.99 and 0.80 to 0.98, respectively). In the 270 images (135 pairs) in which vessel wall measurements were possible, the BNR significantly improved the intraobserver and interobserver correlation coefficients for plaque plus media CSA (0.83 to 0.99 and 0.76 to 0.97, respectively), whereas no influence was observed for external elastic membrane CSA (1.00 to 1.00 and 0.99 to 0.99, respectively). CONCLUSIONS: This study demonstrates the feasibility of this novel algorithm to reduce blood noise, thereby enabling accurate lumen border delineation and providing reproducible measurements of both the lumen and plaque plus media CSAs. Incorporating a digital BNR may serve as an important adjunct to ultra-high-frequency IVUS imaging for improving accurate quantitative evaluation of vessel dimensions. PMID- 11015345 TI - Mechanisms of discordant alternans and induction of reentry in simulated cardiac tissue. AB - BACKGROUND: T-wave alternans, which is associated with the genesis of cardiac fibrillation, has recently been related to discordant action potential duration (APD) alternans. However, the cellular electrophysiological mechanisms responsible for discordant alternans are poorly understood. METHODS AND RESULTS: We simulated a 2D sheet of cardiac tissue using phase 1 of the Luo-Rudy cardiac action potential model. A steep (slope >1) APD restitution curve promoted concordant APD alternans and T-wave alternans without QRS alternans. When pacing was from a single site, discordant APD alternans occurred only when the pacing rate was fast enough to engage conduction velocity (CV) restitution, producing both QRS and T-wave alternans. Tissue heterogeneity was not required for this effect. Discordant alternans markedly increases dispersion of refractoriness and increases the ability of a premature stimulus to cause localized wavebreak and induce reentry. In the absence of steep APD restitution and of CV restitution, sustained discordant alternans did not occur, but reentry could be induced if there was marked electrophysiological heterogeneity. Both discordant APD alternans and preexisting APD heterogeneity facilitate reentry by causing the waveback to propagate slowly. CONCLUSION: Discordant alternans arises dynamically from APD and CV restitution properties and markedly increases dispersion of refractoriness. Preexisting and dynamically induced (via restitution) dispersion of refractoriness independently increase vulnerability to reentrant arrhythmias. Reduction of dynamically induced dispersion by appropriate alteration of electrical restitution has promise as an antiarrhythmic strategy. PMID- 11015346 TI - Effect of renal artery stenting on renal function and size in patients with atherosclerotic renovascular disease. AB - BACKGROUND: Renal artery stenting is widely performed, but little is known about its effectiveness in preserving renal function and size in patients with renovascular disease and chronic renal insufficiency. We studied the effect of renal artery stenting on renal function and size in patients with obstructive renovascular disease and chronic renal insufficiency. METHODS AND RESULTS: Stent deployment was performed in patients with chronic renal insufficiency (creatinine >1.5 mg. dL(-1)) and global renovascular obstruction (bilateral renal artery stenosis or unilateral stenosis in the presence of a solitary or single functional kidney). The effect of renal artery stenting on renal function was assessed by comparing the slopes of the regression lines derived from the reciprocal of serum creatinine versus time plotted before and after stent deployment. Renal size was assessed by serial ultrasound of pole-to-pole kidney length. Stenting was successful in all 61 vessels in 33 patients. Twenty-five patients had complete follow-up (mean 20+/-11 months). Before stent deployment, all patients exhibited a negative slope, indicating progressive renal insufficiency. After stent deployment, the slopes were positive in 18 and less negative in 7 patients. Thus, the mean slope increased from -0.0079 to 0.0043 dL. mg(-1). mo(-1) (P:<0.001). Ultrasonography on 41 kidneys revealed preservation of size, with the kidney length measuring 10.4+/-1.4 cm at baseline and 10.4+/-1.1 cm at last follow-up (P:=NS). Patient survival at 20+/-11 months was 90%. CONCLUSIONS: In patients with chronic renal insufficiency and global obstructive atherosclerotic renovascular disease, renal artery stenting improves or stabilizes renal function and preserves kidney size. PMID- 11015347 TI - Early assessment of myocardial salvage by contrast-enhanced magnetic resonance imaging. AB - BACKGROUND: Myocardial salvage after acute myocardial infarction is defined clinically by early restoration of flow and long-term improvement in contractile function. We hypothesized that contrast-enhanced magnetic resonance imaging (MRI), performed early after myocardial infarction, indexes myocardial salvage. We studied the relationship between the transmural extent of hyperenhancement by contrast-enhanced MRI, restoration of flow, and recovery of function. METHODS AND RESULTS: The left anterior descending coronary artery was occluded in dogs (n=15) for either 45 minutes, 90 minutes, or permanently. Cine and contrast-enhanced MRI were performed 3 days after the procedure; cine MRI was also done 10 and 28 days after the procedure. The transmural extent of hyperenhancement and wall thickening were determined using a 60-segment model. The mean transmural extent of hyperenhancement for the 45-minute occlusion group was 22% of the 90-minute group and 18% of the permanent occlusion group (P:<0.05 for both). The transmural extent of hyperenhancement on day 3 was related to future improvement in both wall thickening score and absolute wall thickening at 10 and 28 days (P:<0.0001 for each). For example, of the 415 segments on day 3 that were dysfunctional and had <25% transmural hyperenhancement, 362 (87%) improved by day 28. Conversely, no segments (0 of 9) with 100% hyperenhancement improved. The transmural extent of hyperenhancement on day 3 was a better predictor of improvement in contractile function than occlusion time (P:<0.0001). CONCLUSIONS: A reduction in the transmural extent of hyperenhancement by contrast-enhanced MRI early after myocardial infarction is associated with an early restoration of flow and future improvement in contractile function. PMID- 11015348 TI - Inhibitory effect of angiotensin II type 2 receptor on coronary arterial remodeling after aortic banding in mice. AB - BACKGROUND: The renin-angiotensin system is thought to be critical for the development of cardiac hypertrophy, whereas the role of the angiotensin II type 2 (AT(2)) receptor in the process is not defined. Using the AT(2) receptor-null (Agtr2-) mouse, we tested the hypothesis that the AT(2) receptor could exert an antigrowth effect in cardiac hypertrophy. METHODS AND RESULTS: Cardiac hypertrophy was induced by suprarenal abdominal aortic banding in 10- to 12-week old Agtr2- and wild-type (Agtr2+) mice for 6 or 12 weeks. Carotid arterial pressure was not different between the strains, although aortic banding increased arterial pressure by approximately 40 mm Hg. Aortic banding increased the heart weight/body-weight ratio and the cross-sectional area of cardiomyocytes by 15%, resulting in comparable cardiomyocyte hypertrophy in the 2 strains. In contrast, coronary arterial thickening and perivascular fibrosis, determined by the media/lumen-area ratio and the collagen/vessel-area ratio, respectively, were 50% greater in Agtr2- than in Agtr2+ mice after banding, whereas these parameters were similar in sham-operated mice. Radioligand binding studies using the whole heart and immunohistochemistry showed that AT(2) receptor expression was limited and localized in the coronary artery and perivascular region. CONCLUSIONS: These results suggest that the AT(2) receptor mediates an inhibitory effect on coronary arterial remodeling, such as medial hypertrophy and perivascular fibrosis in response to pressure overload, and an activation of the renin-angiotensin system. PMID- 11015349 TI - Cardiac-specific overexpression of tumor necrosis factor-alpha causes oxidative stress and contractile dysfunction in mouse diaphragm. AB - BACKGROUND: We have developed a transgenic mouse with cardiac-restricted overexpression of tumor necrosis factor-alpha (TNF-alpha). These mice develop a heart failure phenotype characterized by left ventricular dysfunction and remodeling, pulmonary edema, and elevated levels of TNF-alpha in the peripheral circulation from cardiac spillover. Given that TNF-alpha causes atrophy and loss of function in respiratory muscle, we asked whether transgenic mice developed diaphragm dysfunction and whether contractile losses were caused by oxidative stress or tissue remodeling. METHODS AND RESULTS: muscles excised from transgenic mice and littermate controls were studied in vitro with direct electrical stimulation. Cytosolic oxidant levels were measured with 2', 7' dichlorofluorescin diacetate; emissions of the oxidized product were detected by fluorescence microscopy. Force generation by the diaphragm of transgenic animals was 47% less than control (13.2+/-0. 8 [+/-SEM] versus 25.1+/-0.6 N/cm(2); P:<0.001); this weakness was associated with greater intracellular oxidant levels (P:<0.025) and was partially reversed by 30-minute incubation with the antioxidant N:-acetylcysteine 10 mmol/L (P:<0.01). Exogenous TNF-alpha 500 micromol/L increased oxidant production in diaphragm of wild-type mice and caused weakness that was inhibited by N:-acetylcysteine, suggesting that changes observed in the diaphragm of transgenic animals were mediated by TNF-alpha. There were no differences in body or diaphragm weights between transgenic and control animals, nor was there evidence of muscle injury or apoptosis. CONCLUSIONS: Elevated circulating levels of TNF-alpha provoke contractile dysfunction in the diaphragm through an endocrine mechanism thought to be mediated by oxidative stress. PMID- 11015350 TI - Direct evidence for cytokine involvement in neointimal hyperplasia. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) and interleukin 1 (IL-1) are proximal inflammatory cytokines that stimulate expression of adhesion molecules and induce synthesis of other proinflammatory cytokines. In addition, TNF-alpha and IL-1 influence vascular smooth muscle cell migration and proliferation in vitro. In view of the inflammatory nature of neointimal hyperplasia (NIH), we tested the hypothesis that endogenous TNF-alpha and IL-1 modulate low shear stress-induced NIH. METHODS AND RESULTS: Mice underwent unilateral common carotid artery (CCA) ligation. Low shear stress in the patent ligated CCA has previously been shown to result in remodeling and NIH. Reverse transcriptase-polymerase chain reaction for TNF-alpha and IL-1alpha mRNA demonstrated both TNF-alpha and IL-1alpha mRNA in ligated CCAs, whereas normal and sham-operated CCAs had none. Mice lacking functional TNF-alpha (TNF-/-) developed 14-fold less neointimal area than WT controls (P:<0.05). p80 IL-1 type I receptor knockout (IL-1RI-/-) mice tended to develop less (7-fold, P:>0.05) neointimal area than WT controls. Furthermore, no IL-1alpha mRNA expression was detected in CCAs from TNF-/- mice; however, TNF-alpha mRNA expression was found in the IL-1RI-/- mice. Mice that overexpress membrane-bound TNF-alpha but produce no soluble TNF-alpha display an accentuated fibroproliferative response to low shear stress (P:<0.05). CONCLUSIONS: These results directly demonstrate that TNF-alpha and IL-1 modulate NIH induced by low shear stress. NIH can proceed by way of soluble TNF-alpha independent mechanisms. Specific anti-TNF-alpha and anti-IL-1 therapies may lessen NIH. PMID- 11015351 TI - Confocal microscopic localization of constitutive and heat shock-induced proteins HSP70 and HSP27 in the rat heart. AB - BACKGROUND: Heat-shock treatment of rats elevates expression of heat-shock proteins, which play a role in improving the contractile recovery and reducing infarct size in hearts after ischemic injury. However, the location of these proteins in the heart is unknown. METHODS AND RESULTS: Anesthetized rats were heat-shocked by elevation of body temperature to 42 degrees C to 42.5 degrees C for 15 minutes, followed by 24 hours of recovery. Control and heat-shocked hearts were extirpated and perfused briefly with saline followed by 2% paraformaldehyde in PBS. Confocal immunofluorescence microscopy of control hearts revealed that HSP27 was localized in cardiomyocytes in a pattern reminiscent of Z bands and was colocalized with neuronal markers in somata and axons. No obvious change in HSP27 content or distribution occurred after heat shock. Confocal microscopy revealed little or no HSP70 in control hearts. After heat shock, HSP70 was detected neither in cardiomyocytes nor in neuronal elements within the heart, but HSP70 was abundant in small blood vessels found between the ventricular cardiomyocytes. CONCLUSIONS: Heat shock induces a cell type-specific expression of HSP70 in blood vessels but not myocytes or intrinsic cardiac neurons, suggesting that blood vessels play a primary role in myocardial protection. PMID- 11015352 TI - Roles of P-selectin in inflammation, neointimal formation, and vascular remodeling in balloon-injured rat carotid arteries. AB - BACKGROUND: P-selectin plays key roles in mediating inflammation through promoting adherence of leukocytes to activated platelets and endothelium. This process is one of the initial events in atherosclerosis and restenosis after coronary angioplasty. METHODS AND RESULTS: Using a rat balloon-injury model, we examined the role of P-selectin in vascular inflammatory processes. In the acute phase, immunohistochemistry revealed that P-selectin was intensely expressed on both activated platelets covering the denuded segment and endothelial cells of the inflamed adventitial small vessels. Treatment with an anti-P-selectin monoclonal antibody (MAb) for 8 consecutive days significantly inhibited neointimal formation at day 14 (42% inhibition; P:<0.05), and this effect persisted at day 56 (40% inhibition; P:<0.01) compared with the control group. Vascular shrinking accompanying adventitial fibrosis was also attenuated at day 56. Inhibition of both neointimal formation and vascular shrinking resulted in the lumen area of the anti-P-selectin treatment group being approximately 3 times larger at day 56 than that of the control group. Accumulation of CD45-positive leukocytes in the developing neointima, media, and adventitia at day 8 was significantly inhibited by treatment with the anti-P-selectin MAb. Scanning electron microscopy demonstrated that anti-P-selectin treatment resulted in a less thrombogenic surface of the arterial intima, which featured a pseudoendothelial appearance at day 14 after injury. CONCLUSIONS: These results suggest that inhibition of P-selectin-mediated leukocyte recruitment prevents the development of neointimal formation, adventitial inflammation, and vascular shrinking and promotes pseudoendothelialization by luminal smooth muscle cells. This treatment thus beneficially affects vascular remodeling after balloon injury in rats. PMID- 11015354 TI - Unilateral Vineberg arterial graft with a patency of 30 years. PMID- 11015353 TI - Secondary pulmonary hypertension in chronic heart failure: the role of the endothelium in pathophysiology and management. AB - Pulmonary vascular resistance is frequently elevated in patients with chronic left ventricular failure as a result of dysregulation of vascular smooth muscle tone and structural remodeling. The former is reversible over a period of minutes to days by pharmacological vasodilators, whereas the latter is relatively fixed and may resolve only slowly, over months to years. These abnormalities are due, at least in part, to pulmonary vascular endothelial dysfunction that results in impaired nitric oxide availability and increased endothelin expression. In patients with chronic heart failure, the resulting pulmonary hypertension directly affects right ventricular function and may affect exercise capacity, morbidity, and mortality. New treatment strategies, which include the use of agents that increase nitric oxide availability or oppose the actions of endothelin, may improve the structure and function of the pulmonary vasculature in patients with chronic heart failure. PMID- 11015355 TI - American College of Cardiology/American Heart Association Clinical Competence Statement on Stress Testing. A Report of the American College of Cardiology/American Heart Association/American College of Physicians-American Society of Internal Medicine Task Force on Clinical Competence. PMID- 11015357 TI - Defibrillator disagreement. PMID- 11015356 TI - Visualization of automatic implantable cardioverter-defibrillator patches using electron beam angiography. PMID- 11015358 TI - Multicenter evaluation of epidemiological typing of methicillin-resistant Staphylococcus aureus strains by repetitive-element PCR analysis. The European Study Group on Epidemiological Markers of the ESCMID. AB - Rapid and efficient epidemiologic typing systems would be useful to monitor transmission of methicillin-resistant Staphylococcus aureus (MRSA) at both local and interregional levels. To evaluate the intralaboratory performance and interlaboratory reproducibility of three recently developed repeat-element PCR (rep-PCR) methods for the typing of MRSA, 50 MRSA strains characterized by pulsed field gel electrophoresis (PFGE) (SmaI) analysis and epidemiological data were blindly typed by inter-IS256, 16S-23S ribosomal DNA (rDNA), and MP3 PCR in 12 laboratories in eight countries using standard reagents and protocols. Performance of typing was defined by reproducibility (R), discriminatory power (D), and agreement with PFGE analysis. Interlaboratory reproducibility of pattern and type classification was assessed visually and using gel analysis software. Each typing method showed a different performance level in each center. In the center performing best with each method, inter-IS256 PCR typing achieved R = 100% and D = 100%; 16S-23S rDNA PCR, R = 100% and D = 82%; and MP3 PCR, R = 80% and D = 83%. Concordance between rep-PCR type and PFGE type ranged by center: 70 to 90% for inter-IS256 PCR, 44 to 57% for 16S-23S rDNA PCR, and 53 to 54% for MP3 PCR analysis. In conclusion, the performance of inter-IS256 PCR typing was similar to that of PFGE analysis in some but not all centers, whereas other rep-PCR protocols showed lower discrimination and intralaboratory reproducibility. None of these assays, however, was sufficiently reproducible for interlaboratory exchange of data. PMID- 11015359 TI - Relevance of CagA positivity to clinical course of Helicobacter pylori infection in children. AB - A potential virulence determinant of Helicobacter pylori is the cagA gene product. To determine the relevance of the expression of CagA to the clinical picture and outcome of H. pylori infection in children, we examined 104 consecutive children diagnosed with H. pylori infection. Serum samples were collected to test for the presence of immunoglobulin G (IgG) anti-CagA antibodies. Forty-five patients (43%) had antibodies to the CagA protein (group I), and 59 did not (group II). Seropositive patients had a longer prediagnostic history of abdominal pain (P = 0.02), more severe abdominal pain (defined as ulcer pain) (P = 0.05), a higher prevalence of duodenal ulcer (38 versus 7%; P<0.01), more active chronic gastritis (82 versus 32%; P<0.001), and a higher titer of serum IgG anti-H. pylori antibodies (P<0.001). Ninety percent of the patients were monitored for 27+/-18 months. On multivariate analysis, CagA negative patients had a 3.8-fold-higher chance of achieving a disease-free state than CagA-positive patients (95% confidence interval, 1.5- to 9.5-fold). We conclude that infection with CagA-producing strains of H. pylori is a risk factor for severe clinical disease and ongoing infection. PMID- 11015361 TI - Multicenter evaluation of AMPLICOR and automated COBAS AMPLICOR CT/NG tests for Neisseria gonorrhoeae. AB - The fully automated COBAS AMPLICOR CT/NG and semiautomated AMPLICOR CT/NG tests were evaluated in a multicenter trial for their ability to detect Neisseria gonorrhoeae infections. Test performance compared to that of culturing was evaluated for 2,192 matched endocervical swab and urine specimens obtained from women and for 1, 981 matched urethral swab and urine specimens obtained from men. Culture-negative, PCR-positive specimens that tested positive in a confirmatory PCR test for an alternative target sequence within the N. gonorrhoeae 16S rRNA gene were considered to be true positives. The overall prevalences of gonorrhea were 6.6% in women and 20.1% in men. The COBAS AMPLICOR and AMPLICOR formats yielded concordant results for 98.8% of the specimens and exhibited virtually identical sensitivities and specificities. The results that follow are for the COBAS AMPLICOR format. With the infected patient as the reference standard, the resolved sensitivities of PCR were 92.4% for endocervical swab specimens and 64.8% for female urine specimens. There were no significant differences in these rates between women with and without symptoms. Among symptomatic men, COBAS AMPLICOR sensitivities were 94.1% for urine and 98.1% for urethral swabs; for asymptomatic men, the results were 42.3 and 73.1%, respectively. In comparison, the sensitivities of culturing were 84.8% for endocervical specimens, 92.7% for symptomatic male urethral specimens, and only 46.2% for urethral specimens obtained from asymptomatic men. When PCR results were analyzed as if only a single test had been performed on a single specimen type, the resolved sensitivity was always higher. The resolved specificities of PCR were 99.5% for endocervical swab specimens, 99.8% for female urine specimens, 98.9% for male urethral swab specimens, and 99.9% for male urine specimens. The internal control revealed that 2.1% of specimens were inhibitory when initially tested. Nevertheless, valid results were obtained for 99.2% of specimens because 60.0% of the inhibitory specimens were not inhibitory when a second aliquot was tested. The COBAS AMPLICOR CT/NG test for N. gonorrhoeae exhibited high sensitivity and specificity with urethral swab and urine specimens from men and endocervical swab specimens from women and thus is well suited for diagnosing and screening for N. gonorrhoeae infection. PMID- 11015360 TI - Characterization of enterovirus isolates from patients with heart muscle disease in a selenium-deficient area of China. AB - An association of enterovirus infection with endemic cardiomyopathy (Keshan disease [KD]) and outbreaks of myocarditis in selenium-deficient rural areas of southwestern China has been established. Enteroviruses have been isolated from patients with KD or during outbreaks of myocarditis in last two decades. Six of these isolates grew readily in cell lines (Vero or HEp-2) and were investigated by a novel molecular typing method apart from serotyping and pathogenicity. A neutralization assay identified two isolates from KD as coxsackievirus serotype B2 (CVB2) and two isolates from myocarditis as coxsackievirus serotype B6 (CVB6) but failed to type the remaining two isolates, also from myocarditis. Direct nucleotide sequencing of reverse transcription-PCR products amplified from the 5' nontranslated region (5'NTR) of these viruses confirmed that they belong to a phylogenetic cluster consisting of coxsackie B-like viruses, including some echovirus serotypes. Sequence analysis of the coding region for viral capsid protein VP1 showed that two isolates serotyped as CVB2 have the highest amino acid sequence homology with CVB2 and that the remaining four isolates, two CVB6 and the two unknown serotypes, are most closely related to the sequence of CVB6. Sequences among these isolates varied from 82.3 to 99% in the 5'NTR and from 69 to 99% in VP1, indicating no cross contamination. The pathogenicity of these viruses in adult and suckling mice was assessed. None caused pathologic changes in the hearts of adult MF-1 or SWR mice, although pancreatitis was evident. However, the four CVB6-like viruses caused death in suckling mice, similar to a virulent coxsackievirus group B3 laboratory strain. In conclusion, the sequence data confirm that coxsackievirus group B serotypes are predominant in the region in which KD is endemic and may be the etiological agents in outbreaks of myocarditis. VP1 genotyping of enteroviruses is accurate and reliable. Animal experiments indicate that isolates may differ in pathogenicity. PMID- 11015362 TI - Diarrheagenic Escherichia coli and acute and persistent diarrhea in returned travelers. AB - To determine the role of diarrheagenic Escherichia coli in acute and persistent diarrhea in returned travelers, a case control study was performed. Enterotoxigenic E. coli (ETEC) was detected in stool samples from 18 (10.7%) of 169 patients and 4 (3.7%) of 108 controls. Enteroaggregative E. coli (EAggEC) was detected in 16 (9.5%) patients and 7 (6.5%) controls. Diffuse adherent E. coli strains were commonly present in both patients (13%) and controls (13.9). Campylobacter and Shigella species were the other bacterial enteropathogens most commonly isolated (10% of patients, 2% of controls). Multivariate analysis showed that the presence of ETEC was associated with acute diarrhea (odds ratio [OR], 6.7; 95% confidence interval [CI], 1.5 to 29.1; P = 0.005), but not with persistent diarrhea (OR, 1.6; 95% CI, 0.4 to 7.4). EAggEC was significantly more often present in patients with acute diarrhea than in controls (P = 0.009), but no significant association remained after multivariate analysis. ETEC and EAggEC are frequently detected in returned travelers with diarrhea. The presence of ETEC strains is associated with acute but not with persistent diarrhea. PMID- 11015363 TI - Brachyspira aalborgi infection diagnosed by culture and 16S ribosomal DNA sequencing using human colonic biopsy specimens. AB - In this study we report on the isolation and characterization of the intestinal spirochete Brachyspira aalborgi using human mucosal biopsy specimens taken from the colon of a young adult male with intestinal spirochetosis. A selective medium, containing 400 microg of spectinomycin/ml and 5 microg of polymyxin/ml was used for the isolation procedure. A high degree of similarity, in terms of phenotypic properties and 16S ribosomal DNA sequence, was observed between the isolated strain, named W1, and the type strain, 513A, of B. aalborgi. A similarity of 99.7% in the nucleotide sequence was found between W1 and 513A(T), based on the almost-complete gene. A short segment of the 16S rRNA gene was amplified by PCR using genetic material enriched from paraffin-embedded biopsy specimens, which were taken from the patient on two occasions. The products showed 16S rRNA gene sequences virtually identical to that of strain 513A(T) in the actual region. Immunohistochemistry was performed on the colonic biopsy specimens with a polyclonal antibody raised against an intestinal spirochete isolated in a previous case of human intestinal spirochetosis. The antibody reacted strongly with the spirochete on the luminal epithelium. No immune reaction was seen within or below the surface epithelium. Routine histology did not reveal signs of colitis. Electron microscopy showed spirochetes attached end on to the colonic mucosal surface. The isolate grew poorly on a commonly used selective medium for intestinal spirochetes, which may explain previous failures to isolate B. aalborgi. PMID- 11015364 TI - Serodiagnosis of Louse-Borne relapsing fever with glycerophosphodiester phosphodiesterase (GlpQ) from Borrelia recurrentis. AB - Human louse-borne relapsing fever occurs in sporadic outbreaks in central and eastern Africa that are characterized by significant morbidity and mortality. Isolates of the causative agent, Borrelia recurrentis, were obtained from the blood of four patients during a recent epidemic of the disease in southern Sudan. The glpQ gene, encoding glycerophosphodiester phosphodiesterase, from these isolates was sequenced and compared with the glpQ sequences obtained from other relapsing-fever spirochetes. Previously we showed that GlpQ of Borrelia hermsii is an immunogenic protein with utility as a serological test antigen for discriminating tick-borne relapsing fever from Lyme disease. In the present work, we cloned and expressed the glpQ gene from B. recurrentis and used recombinant GlpQ in serological tests. Acute- and convalescent-phase serum samples obtained from 42 patients with louse-borne relapsing fever were tested with an indirect immunofluorescence assay (IFA) and an enzyme-linked immunosorbent assay (ELISA) that used whole cells of B. recurrentis and with immunoblotting to whole-cell lysates of the spirochete and Escherichia coli producing recombinant GlpQ. The geometric mean titers of the acute- and convalescent-phase serum samples measured by IFA were 1:83 and 1:575, respectively. The immunoblot analysis identified a high level of reactivity and seroconversion to GlpQ, and the assay was more sensitive than the whole-cell IFA and ELISA using purified, recombinant histidine tagged GlpQ. Serum antibodies to GlpQ and other antigens persisted for 27 years in one patient. We conclude that assessment of anti-GlpQ antibodies will allow serological confirmation of louse-borne relapsing fever and determination of disease prevalence. PMID- 11015366 TI - Evaluation of the MicroScan rapid neg ID3 panel for identification of Enterobacteriaceae and some common gram-negative nonfermenters. AB - The MicroScan Rapid Neg ID3 panel (Dade Behring, Inc., West Sacramento, Calif.) is designed for the identification of gram-negative bacilli. We evaluated its ability to accurately identify Enterobacteriaceae that are routinely encountered in a clinical laboratory and glucose nonfermenting gram-negative bacilli. Using 511 stock cultures that were maintained at -70 degrees C and passaged three times before use, we inoculated panels according to the manufacturer's instructions and processed them in a Walk/Away instrument using version 22.01 software. The time to identification was 2 h and 30 min. All panel identifications were compared to reference identifications previously determined by conventional tube biochemicals. At the end of the initial 2.5-h incubation period, 405 (79.3%) identifications were correct. An additional 49 (9.6%) isolates were correctly identified after required additional off-line biochemical tests were performed. Thus, at 24 h, 88.8% of the 511 strains tested were correctly identified. Twenty two (4.3%) were identified to the genus level only. Twenty-six (5.1%) strains were misidentified. Because the system is based on fluorogenics, there are no conventional tests readily available with which to compare possibly incorrect reactions. Of the 28 Salmonella strains that were tested, 5 were incorrectly reported. The 21 remaining errors were scattered among the genera tested. Testing on nine strains gave a result of "no identification" (very rare biotype). The Rapid Neg ID3 panel in this study approached 89% accuracy for the identification of gram-negative organisms encountered in the hospital laboratory. PMID- 11015365 TI - Spoligotyping and polymorphic GC-rich repetitive sequence fingerprinting of mycobacterium tuberculosis strains having few copies of IS6110. AB - Several genetic loci have been utilized to genotype isolates of Mycobacterium tuberculosis. A shortcoming of the most commonly used method, IS6110 fingerprinting, is that it does not adequately discriminate between isolates having few copies of IS6110. This study was undertaken to compare pTBN12 fingerprinting of polymorphic GC-rich repetitive sequence genes and spoligotyping of the direct repeat locus as secondary typing procedures for M. tuberculosis isolates having fewer than six copies of IS6110. A total of 88 isolates (100% of the isolates with fewer than six copies of IS6110 isolated in Arkansas during 1996 and 1997) were included in this study. Among the 88 isolates, 34 different IS6110 patterns were observed, 10 of which were shared by more than 1 isolate, involving a total of 64 isolates. The 64 isolates were subdivided into 13 clusters (containing 37 isolates) and 27 unique isolates based on a combination of IS6110 and pTBN12 fingerprinting and into 11 clusters (containing 51 isolates) and 13 unique isolates based on a combination of IS6110 fingerprinting and spoligotyping. Identical spoligotypes were found among isolates having different IS6110 patterns, as well as among isolates showing different pTBN12 patterns. In contrast, all isolates that had different IS6110 patterns were found to be unique by pTBN12 typing. The clustering rate was 73, 58, and 42%, respectively, for IS6110 fingerprinting alone, IS6110 fingerprinting and spoligotyping combined, and IS6110 and pTBN12 combined fingerprinting. The data indicate that the pTBN12 method has greater discriminating power among low-copy-number isolates than does spoligotyping. PMID- 11015367 TI - Genotyping of hepatitis C virus isolates using CLIP sequencing. AB - Determination of hepatitis C virus (HCV) genotypes and subtypes has become increasingly important for the clinical management and prognosis of HCV infections. The aim of the present study was to assess the specificity and reliability of a newly developed, commercially available HCV genotyping kit (TRUGENE HCV 5'NC genotyping kit). This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5' noncoding region (5'NCR). HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5'NC genotyping kit and DNA enzyme immunoassay (DEIA). Typing results obtained by both methods were in complete concordance in 91% of the cases. HCV RNA from the samples with discordant genotype assignment in both assays was additionally amplified with primers from the HCV core and NS5B regions. Phylogenetic analysis of the obtained sequences supported the results obtained from DEIA in six cases and CLIP sequencing in two cases. In the former six cases, the TRUGENE HCV 5'NC genotyping kit could not correctly differentiate between subtypes of genotypes 1 and 2 due to the high conservation of the 5'NCR. However, since there was not any misclassification between HCV genotypes 1 and non-1 types, the results obtained with this system are, in general, reliable and can be used in clinical practice. The TRUGENE HCV 5'NC genotyping kit in our hands proved to be a fast and convenient technique that might be an attractive option for HCV genotyping in laboratories already using the Roche AMPLICOR HCV test for diagnostic reverse transcription-PCR. PMID- 11015368 TI - Detection of trichomonosis in vaginal and urine specimens from women by culture and PCR. AB - Vaginal trichomonosis is a highly prevalent infection which has been associated with human immunodeficiency virus acquisition and preterm birth. Culture is the current "gold standard" for diagnosis. As urine-based testing using DNA amplification techniques becomes more widely used for other sexually transmitted diseases (STDs) such as gonorrhea and chlamydia, a similar technique for trichomonosis would be highly desirable. Women attending an STD clinic for a new complaint were screened for Trichomonas vaginalis by wet-preparation (wet-prep) microscopy and culture and for the presence of T. vaginalis DNA by specific PCR of vaginal and urine specimens. The presence of trichomonosis was defined as the detection of T. vaginalis by direct microscopy and/or culture from either vaginal samples or urine. The overall prevalence of trichomonosis in the population was 28% (53 of 190). The sensitivity and specificity of PCR using vaginal samples were 89 and 97%, respectively. Seventy-four percent (38 of 51) of women who had a vaginal wet prep or vaginal culture positive for trichomonads had microscopic and/or culture evidence of the organisms in the urine. Two women were positive for trichomonads by wet prep or culture only in the urine. The sensitivity and specificity of PCR using urine specimens were 64 and 100%, respectively. These results indicate that the exclusive use of urine-based detection of T. vaginalis is not appropriate in women. PCR-based detection of T. vaginalis using vaginal specimens may provide an alternative to culture. PMID- 11015369 TI - Rapid differentiation of fermentative from nonfermentative gram-negative bacilli in positive blood cultures by an impedance method. AB - Rapid differentiation of fermentative gram-negative bacilli (fermenters) from nonfermentative gram-negative bacilli (nonfermenters) in positive blood cultures may help physicians to narrow the choice of appropriate antibiotics for empiric treatment. An impedance method for direct differentiation of fermenters from nonfermenters was investigated. The bacterial suspensions (or positive culture broths containing gram-negative bacteria) were inoculated into the module wells of a Bactometer (bioMerieux, Inc., Hazelwood, Mo.) containing 1 ml of Muller Hinton broth. The inoculated modules were incubated at 35 degrees C, and the change in impedance in each well was continuously monitored. The amount of time required to cause a series of significant deviations from baseline impedance values was defined as the detection time (DT). The percent change of impedance was defined as the change of impedance at the time interval from DT to DT plus 1 h. After testing 857 strains of pure cultures (586 strains of fermenters and 271 strains of nonfermenters), a breakpoint (2.98%) of impedance change was obtained by discriminant analysis. Strains displaying impedance changes of greater than 2.98% were classified as fermenters; the others were classified as nonfermenters. By using this breakpoint, 98.6% (340 of 345) of positive blood cultures containing fermenters and 98% (98 of 100) of positive blood cultures containing nonfermenters were correctly classified. The impedance method was simple, and the results were normally available within 2 to 4 h after direct inoculation of positive blood culture broths. PMID- 11015370 TI - Elevated phenotypic switching and drug resistance of Candida albicans from human immunodeficiency virus-positive individuals prior to first thrush episode. AB - Strains of Candida albicans obtained from human immunodeficiency virus (HIV) positive individuals prior to their first episode of oral thrush were already in a high-frequency mode of switching and were far more resistant to a number of antifungal drugs than commensal isolates from healthy individuals. Switching in these isolates also had profound effects both on susceptibility to antifungal drugs and on the levels of secreted proteinase activity. These results suggest that commensal strains colonizing HIV-positive individuals either undergo phenotypic alterations or are replaced prior to the first episode of oral thrush. They also support the suggestion that high-frequency phenotypic switching functions as a higher-order virulence trait, spontaneously generating in colonizing populations variants with alterations in a variety of specific virulence traits. PMID- 11015371 TI - Increased sensitivity of the BACTEC 460 mycobacterial radiometric broth culture system does not decrease the number of respiratory specimens required for a definitive diagnosis of pulmonary tuberculosis. AB - The BACTEC 460 radiometric mycobacterial broth culture system has consistently demonstrated faster and increased recovery of Mycobacterium tuberculosis from respiratory specimens of patients with pulmonary tuberculosis than conventional culture methods. We thus questioned whether three sputa were still necessary to definitively diagnose pulmonary tuberculosis if the BACTEC radiometric culture system were in use. We performed a retrospective analysis of 430 sequential respiratory specimens submitted from 143 patients and from which M. tuberculosis had been recovered by in vitro culture and simultaneously assessed the diagnostic yield of acid-fast smear in this same cohort. M. tuberculosis was recovered from the first specimen for 117 (82%) of the 143 patients, from the second for 14 patients (10%; cumulative rate, 92%), and from the third for 12 patients (8%; cumulative rate, 100%). With the exception of those for bronchial brushings, recovery rates of M. tuberculosis were comparable for all respiratory specimen types (expectorated sputum, induced sputum, tracheal aspirates, bronchoalveolar lavage fluids). Only 46 (32%) of these 143 patients had acid-fast bacilli detected in smears; acid-fast bacilli were detected in the first submitted specimen for 44 patients (96%) and in the second for the remaining 2 patients (4%; cumulative rate, 100%). Culture- or smear-positive rates for sequential specimens obtained from AIDS patients were comparable to those for non-AIDS patients. Overall, the diagnostic culture yield of sequentially submitted specimens was not different from previously published studies in which the BACTEC radiometric culture system had not been used. Despite the documented enhanced ability of the BACTEC 460 radiometric mycobacterial culture system to recover M. tuberculosis more often and faster than conventional methods, three sequential respiratory specimens (regardless of type) were still necessary to definitively diagnose pulmonary tuberculosis. PMID- 11015372 TI - Use of a repetitive DNA probe to type clinical and environmental isolates of Aspergillus flavus from a cluster of cutaneous infections in a neonatal intensive care unit. AB - Aspergillus flavus is second to A. fumigatus as a cause of invasive aspergillosis, but no standard method exists for molecular typing of strains from human sources. A repetitive DNA sequence cloned from A. flavus and subcloned into a pUC19 vector, pAF28, was used to type 18 isolates from diverse clinical, environmental, and geographic sources. The restriction fragment length polymorphisms generated with EcoRI- or PstI-digested genomic DNA and probed with digoxigenin-labeled pAF28 revealed complete concordance between patterns. Eighteen distinct fingerprints were observed. The probe was used to investigate two cases of cutaneous A. flavus infection in low-birth-weight infants in a neonatal intensive care unit (NICU). Both infants were transported by the same ambulance and crew to the NICU on the same day. A. flavus strains of the same genotype were isolated from both infants, from a roll of tape used to fasten their umbilical catheters, from a canvas bag used to store the tape in the ambulance, and from the tape tray in the ambulance isolette. These cases highlight the need to consider exposures in critically ill neonates that might occur during their transport to the NICU and for stringent infection control practices. The hybridization profiles of strains from a second cluster of invasive A. flavus infections in two pediatric hematology-oncology patients revealed a genotype common to strains from a definite case patient and a health care worker. A probable case patient was infected with a strain with a genotype different from that of the strain from the definite case patient but highly related to that of an environmental isolate. The high degree of discrimination and reproducibility obtained with the pAF28 probe underscores its utility for typing clinical and environmental isolates of A. flavus. PMID- 11015373 TI - Comparative immunoglobulin G antibody profiles between mother and child (CGMC test) for early diagnosis of congenital toxoplasmosis. AB - Early diagnosis of congenital toxoplasmosis is rendered difficult when specific immunoglobulin M (IgM) and/or IgA antibodies are absent in the blood of the newborn infant. Since maternal IgG antibodies can cross the placenta, determination of IgG antibodies in newborn infants has hitherto not been used routinely for the diagnosis of congenital infection. The aim of this study was to assess the diagnostic usefulness of an immunoblot assay which compares the early IgG profiles between the mother and her child (comparative IgG profile between mother and child; CGMC test) directed against a total cell lysate of Toxoplasma gondii tachyzoites. Serum samples from 97 newborn infants at risk of toxoplasma infection were obtained from umbilical cord blood at birth or postnatally until 3 months of life and were directly compared with serum samples from the respective mothers. Congenital toxoplasmosis was diagnosed only when IgG-reactive protein bands that were present in any newborn serum samples were absent in the corresponding maternal serum sample. Congenital infection was defined by conventional serological assays when IgM and/or IgA antibodies were present in newborn infant blood or when IgG titers rose within the first 12 months or were persistently stable for more than 8 months. Using these criteria, congenital infection was definitely confirmed in 11 cases. Three additional cases were diagnosed based on indicative data. The CGMC test, which was performed without knowledge of the results of conventional serologal assays, had sensitivity and specificity of 82.4 and 93.0%, respectively, and positive and negative predictive values of 73.7 and 95.7%, respectively. When true positives and true negatives were considered, the comparative IgG profile had a ratio of 90.9% true results. The CGMC test thus is useful as an additional assay for the rapid diagnosis of congenital toxoplasmosis when paired serum samples from mother and child are available. PMID- 11015374 TI - 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates. AB - Some bacteria are difficult to identify with phenotypic identification schemes commonly used outside reference laboratories. 16S ribosomal DNA (rDNA)-based identification of bacteria potentially offers a useful alternative when phenotypic characterization methods fail. However, as yet, the usefulness of 16S rDNA sequence analysis in the identification of conventionally unidentifiable isolates has not been evaluated with a large collection of isolates. In this study, we evaluated the utility of 16S rDNA sequencing as a means to identify a collection of 177 such isolates obtained from environmental, veterinary, and clinical sources. For 159 isolates (89.8%) there was at least one sequence in GenBank that yielded a similarity score of > or =97%, and for 139 isolates (78.5%) there was at least one sequence in GenBank that yielded a similarity score of > or =99%. These similarity score values were used to defined identification at the genus and species levels, respectively. For isolates identified to the species level, conventional identification failed to produce accurate results because of inappropriate biochemical profile determination in 76 isolates (58.7%), Gram staining in 16 isolates (11.6%), oxidase and catalase activity determination in 5 isolates (3.6%) and growth requirement determination in 2 isolates (1.5%). Eighteen isolates (10.2%) remained unidentifiable by 16S rDNA sequence analysis but were probably prototype isolates of new species. These isolates originated mainly from environmental sources (P = 0.07). The 16S rDNA approach failed to identify Enterobacter and Pantoea isolates to the species level (P = 0.04; odds ratio = 0.32 [95% confidence interval, 0.10 to 1.14]). Elsewhere, the usefulness of 16S rDNA sequencing was compromised by the presence of 16S rDNA sequences with >1% undetermined positions in the databases. Unlike phenotypic identification, which can be modified by the variability of expression of characters, 16S rDNA sequencing provides unambiguous data even for rare isolates, which are reproducible in and between laboratories. The increase in accurate new 16S rDNA sequences and the development of alternative genes for molecular identification of certain taxa should further improve the usefulness of molecular identification of bacteria. PMID- 11015375 TI - Comparison of Danish isolates of Salmonella enterica serovar enteritidis PT9a and PT11 from hedgehogs (Erinaceus europaeus) and humans by plasmid profiling and pulsed-field gel electrophoresis. AB - During the years 1994 to 1998, 10 strains of Salmonella enterica serovar Enteritidis phage type 11 (PT11) and 6 PT9a strains were isolated from Danish hedgehogs, together with 7 strains that did not yield phage susceptibility patterns conforming with any known phage type (routine dilution no conformity [RDNC]). From 1995 to 1998, five Danish patients were reported infected with serovar Enteritidis PT11 and two with PT9a. All serovar Enteritidis PT11, PT9a, and RDNC isolates from hedgehogs and humans were analyzed by pulsed-field gel electrophoresis (PFGE), plasmid profiling, and restriction fragment length polymorphism (RFLP) of plasmids. By use of S1 nuclease and HindIII, the PT11 and PT9a isolates had identical plasmid profiles and RFLP patterns, which differed from the RDNC profiles. The PFGE profiles were identical for all serovar Enteritidis PT11 and PT9a strains from hedgehogs, four of five human strains of serovar Enteritidis PT11, and two human strains of serovar Enteritidis PT9a, irrespective of restriction enzyme, whereas the last human strain deviated slightly when NotI was used but not when XbaI or SpeI was used. The results indicate that serovar Enteritidis PT9a and PT11 are closely related and that PT11 and PT9a from Danish hedgehogs and humans belong to the same clonal lineage. PMID- 11015377 TI - Helicobacter pylori: clonal population structure and restricted transmission within families revealed by molecular typing. AB - Helicobacter pylori infects up to 50% of the human population worldwide. The infection occurs predominantly in childhood and persists for decades or a lifetime. H. pylori is believed to be transmitted from person to person. However, tremendous genetic diversity has been reported for these bacteria. In order to gain insight into the epidemiological basis of this phenomenon, we performed molecular typing of H. pylori isolates from different families. Fifty-nine H. pylori isolates from 27 members of nine families were characterized by using restriction fragment length polymorphism analysis of five PCR-amplified genes, by pulsed-field gel electrophoresis (PFGE) of chromosomal DNA, and by vacA and cagA genotyping. The 16S rRNA gene exhibited little allelic variation, as expected for a unique bacterial species. In contrast, the vacA, flaA, ureAB, and lspA-glmM genes were highly polymorphic, with a mean genetic diversity of 0.83, which exceeds the levels recorded for all other bacterial species. In conjunction with PFGE, 59 H. pylori isolates could be differentiated into 21 clonal types. Each individual harbored only one clone, occasionally with a clonal variant. Identical strains were always found either between siblings or between a mother and her children. Statistical analysis revealed clonality of population structure in all isolates. The results of this study suggest the possible coexistence of a large array of clonal lineages that are evolving in each individual in isolation from one another. Transmission appears to occur primarily from mother to child and perhaps between siblings. PMID- 11015376 TI - Molecular surveillance of European quinolone-resistant clinical isolates of Pseudomonas aeruginosa and Acinetobacter spp. using automated ribotyping. AB - Nosocomial isolates of Pseudomonas aeruginosa and Acinetobacter spp. exhibit high rates of resistance to antibiotics and are often multidrug resistant. In a previous study (D. Milatovic, A. Fluit, S. Brisse, J. Verhoef, and F. J. Schmitz, Antimicrob. Agents Chemother. 44:1102-1107, 2000), isolates of these species that were resistant to sitafloxacin, a new advanced-generation fluoroquinolone with a high potency and a broad spectrum of antimicrobial activity, were found in high proportion in 23 European hospitals. Here, we investigate the clonal diversity of the 155 P. aeruginosa and 145 Acinetobacter spp. sitafloxacin-resistant isolates from that study by automated ribotyping. Numerous ribogroups (sets of isolates with indistinguishable ribotypes) were found among isolates of P. aeruginosa (n = 34) and Acinetobacter spp. (n = 16), but the majority of the isolates belonged to a limited number of major ribogroups. Sitafloxacin-resistant isolates (MICs > 2 mg/liter, used as a provisional breakpoint) showed increased concomitant resistance to piperacillin, piperacillin-tazobactam, ceftriaxone, ceftazidime, amikacin, gentamicin, and imipenem. The major ribogroups were repeatedly found in isolates from several European hospitals; these isolates showed higher levels of resistance to gentamicin and imipenem, and some of them appeared to correspond to previously described multidrug-resistant international clones of P. aeruginosa (serotype O:12) and Acinetobacter baumannii (clones I and II). Automated ribotyping, when used in combination with more discriminatory typing methods, may be a convenient library typing system for monitoring future epidemiological dynamics of geographically widespread multidrug-resistant bacterial clones. PMID- 11015378 TI - Evaluation of D-xylose and 1% methyl-alpha-D-glucopyranoside fermentation tests for distinguishing Enterococcus gallinarum from Enterococcus faecium. AB - To determine the validity of the rapid xylose and methyl-alpha-D-glucopyranoside (MDG) fermentation tests in distinguishing Enterococcus gallinarum from Enterococcus faecium, 156 well-characterized clinical isolates of enterococci (55 E. gallinarum, 91 E. faecium, and 10 Enterococcus faecalis isolates) known to be of different clones were examined in a blinded fashion. Species identification was confirmed by PCR of the ddl ligase genes of E. faecium and E. faecalis and the vanC1 gene of E. gallinarum. Xylose tests were performed with D-xylose tablets by using a heavy bacterial suspension and were interpreted after 2 h of incubation. Standard MDG fermentation tests were read after 24 h of incubation. The xylose fermentation test had a sensitivity of 98% (54 of 55) and a specificity of 99% (100 of 101) in distinguishing E. gallinarum from E. faecium and E. faecalis. The standard MDG test had a sensitivity of 100% (55 of 55) and a specificity of 95% (96 of 101) after 24 h. The xylose fermentation test is a simple method, easily incorporated into laboratory protocols, that distinguishes E. gallinarum from E. faecium with high sensitivity and specificity in 2 h. The standard MDG test has high sensitivity and can be useful in ruling out the presence of E. gallinarum but requires overnight incubation. PMID- 11015379 TI - Genotypic determination of Mycobacterium tuberculosis antibiotic resistance using a novel mutation detection method, the branch migration inhibition M. tuberculosis antibiotic resistance test. AB - A novel method for the detection of any alteration within a defined sequence has recently been demonstrated (A. Lishanski, N. Kurn, and E. F. Ullman, Nucleic Acids Res. 28:E42, 2000; A. Lishanski, Clin. Chem. 46:9, 2000). Essential to this method are the generation of partial duplexes that are capable of forming four stranded structures and the ability to detect inhibition of branch migration in these structures (I. G. Panyutin and P. Hsieh, J. Mol. Biol. 230:413-424, 1993). Inhibition of branch migration indicates the presence of sequence alteration. This mutation detection method, termed branch migration inhibition (BMI), is suitable for the detection of drug resistance in M. tuberculosis, which is frequently associated with multiple mutations within known genes. We describe the genotypic determination of the rifampin (RMP) and pyrazinamide (PZA) susceptibilities of M. tuberculosis isolates, using BMI coupled with the luminescence oxygen channeling immunoassay (LOCI) (E. F. Ullman et al., Proc. Natl. Acad. Sci. USA 91:5426-5430, 1994). RMP and PZA resistances are associated with multiple mutations within the rpoB and pncA genes, respectively. M. tuberculosis genomic DNA samples prepared from 46 clinical isolates were used for genotypic determination of RMP resistance in a "blind study." Similarly, PZA resistance was determined using genomic DNA samples prepared from 37 clinical isolates. Full agreement of the genotypic and phenotypic determinations of drug susceptibility was demonstrated. RMP susceptibility determination directly from cells of 10 clinical isolates grown in culture was also demonstrated. The genotypic result of only 1 out of 10 isolates did not agree with the phenotypic susceptibility testing result. Sequence analysis of the rpoB gene of this clinical isolate revealed a single base substitution, most likely a silent point mutation. The new BMI-LOCI mutation detection method is a rapid and accurate procedure for the genotypic determination of the RMP and PZA susceptibilities of M. tuberculosis clinical isolates. BMI can also be detected by using commercially available automated enzyme-linked immunosorbent assay plate formats (Lishanski et al., Nucleic Acids Res. 28:E42, 2000). PMID- 11015380 TI - Pneumococcal pspA sequence types of prevalent multiresistant pneumococcal strains in the United States and of internationally disseminated clones. AB - In a recent genotypic survey of beta-lactam-resistant pneumococci recovered in different areas of United States during 1997, eight clonal types that each represented 3 to 40 isolates accounted for 134 of 144 isolates (G. Gherardi, C. Whitney, R. Facklam, and B. Beall, J. Infect. Dis. 181:216-229, 2000). We determined the degree of pspA gene diversity among these 134 isolates and for 11 previously characterized internationally disseminated multiresistant strains. Thirty-four different pspA restriction profiles were determined for an amplicon encompassing the variable portion of the structural gene that encodes the surface exposed domain of PspA and a variable-length proline-rich putative cell wall associated domain. These restriction profiles closely correlated with those of 33 different pspA sequence types of an approximately 230-residue region corresponding to residues 182 to 410 of the strain Rx1 PspA. These residues encompass a 100-residue clade-defining region known to contain cross-protective epitopes for which 17 sequence types were found. Distinct, conserved pspA sequence types were found for the majority of strains within seven of the eight U.S. clonal types assessed, while one pulsed-field gel electrophoresis type was represented by isolates of three distinct PspA clades. Sequence typing of pspA provides an added level of specificity in the subtyping of isolates and is a necessary first step in determining the components needed in a PspA vaccine which could elicit effective cross-protective coverage. PMID- 11015382 TI - Evaluation of amplified fragment length polymorphism analysis for inter- and intraspecific differentiation of Mycobacterium bovis, M. tuberculosis, and M. ulcerans. AB - The usefulness of amplified fragment length polymorphism (AFLP) analysis was evaluated for the discrimination of Mycobacterium bovis (17 strains), M. tuberculosis (15 strains), and M. ulcerans (12 strains) at the inter- and intraspecific level. The AFLP technique is a whole-genome coverage genotypic fingerprinting method based on the selective PCR amplification of modified restriction fragments obtained through a double enzymatic digest and subsequent ligation of double-stranded restriction site-specific adapter oligonucleotides. Selective amplification of ApaI/TaqI templates with primer combination A02-T02 (both having an additional C at their 3' end) generated autoradiographic AFLP fingerprints that were grouped by numerical analysis in two main AFLP clusters allowing clear separation of M. ulcerans (cluster I) from the M. tuberculosis complex members M. bovis and M. tuberculosis (cluster II). Calculation of similarities using the band-based Dice correlation coefficient instead of the Pearson product-moment correlation coefficient revealed a further subgrouping in cluster II. The two resulting subclusters corresponded with the phenotypic identity of M. bovis and M. tuberculosis, respectively, and could also be visually identified by two AFLP marker bands. Because of the relatively low degree of genotypic variation among the AFLP band patterns of the latter two taxa, no correlation could be found with previously reported molecular typing data or with geographical origin. The use of primer combination A02-T01 (the latter having an A as selective base) did not increase the resolving power within the M. tuberculosis complex but resulted in a visual subgrouping of the M. ulcerans strains that was not observed with primer combination A02-T02. Based on the presence or absence of a single AFLP marker band, the M. ulcerans isolates could be unambiguously classified in two continental types corresponding with the African and Australian origin of the strains, respectively. In conclusion, the radioactive AFLP method proved to be a reproducible and reliable taxonomic tool for the differentiation of the three mycobacterial species under study and also demonstrated its potential use for typing of M. ulcerans strains when employing multiple primer combinations. PMID- 11015381 TI - Detection of borreliacidal antibodies in dogs after challenge with Borrelia burgdorferi-infected ixodes scapularis ticks. AB - Detection of borreliacidal antibodies is an accurate serodiagnostic test for confirmation of Lyme disease in humans. In this study, 13 pathogen-free beagles, 12 to 26 weeks old, were infected with Borrelia burgdorferi by tick challenge. Dogs were monitored for clinical signs and symptoms of Lyme disease along with borreliacidal antibody production against B. burgdorferi sensu stricto isolates 297 and 50772. Ten (77%) dogs developed lameness in one or more legs within 210 days after attachment of Ixodes scapularis ticks. Eight (80%) of the lame animals had concurrent fever of > or =38 degrees C. Spirochetes were also recovered from the skin and joints of 12 (92%) dogs, but rarely from other organs. Borreliacidal antibodies against B. burgdorferi isolate 297 were detected in only four (31%) dogs, and the levels of killing antibodies remained low for the duration of the infection. In contrast, borreliacidal antibodies against B. burgdorferi isolate 50772 were detected in 13 (100%) dogs within 21 days of infection. Furthermore, the borreliacidal antibody levels correlated with the severity of B. burgdorferi infection. Detection of borreliacidal antibodies, especially against B. burgdorferi isolate 50772, is also a reliable serodiagnostic test for detection of Lyme disease in dogs. PMID- 11015383 TI - Genotyping of rotaviruses in environmental water and stool samples in Southern Switzerland by nucleotide sequence analysis of 189 base pairs at the 5' end of the VP7 gene. AB - Stool specimens from children (<4 years old) with diarrhea were collected over a 1-year period in Ticino (southern region of Switzerland). During the same period, environmental samples were collected from surface waters in the proximity of major water treatment plants. From treatment plants, samples were collected from the raw sewage and before the release of the treated water. From rivers, samples were collected before and after receiving the treated waters. A single-step reverse transcription (RT)-PCR amplification of the entire VP7 gene from extracted double-stranded RNA was developed. For the water samples, a further nested PCR was necessary to increase sensitivity. All amplified viral products were sequenced, and the sequence profile was compared to that of the VP7 genes of human and animal rotaviruses from GenBank. Rotavirus strains are characterized by outer capsid proteins G (glycoprotein) and P (protease-cleaved protein). Correct G genotyping of viral sequences from stool and water samples was possible by analyzing only 189 bp at the 5' end of the VP7 gene. In the Ticino region, the most predominant G genotype among clinical and water samples was G1. Genotypes G2 and G4 were found only among clinical samples. We also detected rotavirus G1-type sequences in feces from a healthy adult. This finding corroborates the hypothesis that healthy adults act as potential reservoirs for the spread of rotavirus in the environment. In our experiments, this RT-PCR-based method for rotavirus genotyping has proven to be a useful tool for epidemiological investigations. PMID- 11015384 TI - Comparison of phenotypic and genotypic methods for pyrazinamide susceptibility testing with Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis converts pyrazinamide to its active form by using the enzyme pyrazinamidase. This enzyme is coded for on the pncA gene, and mutations in the pncA gene result in absence of active enzyme, conferring resistance to the drug pyrazinamide. We investigated 27 strains of Mycobacterium tuberculosis suspected of being multidrug resistant. Each isolate was tested for susceptibility to pyrazinamide by the BACTEC 460TB method, and 19 were pyrazinamide resistant. The presence of active pyrazinamidase enzyme was sought by using the Wayne assay, which was positive in all of the sensitive isolates and four of the resistant isolates. The pncA gene was amplified by PCR, and mutations were sought by single-strand conformation polymorphism (SSCP) analysis. We identified four isolates which were phenotypically resistant to pyrazinamide, but which had active pyrazinamide enzyme on the Wayne assay and normal pncA gene SSCP. MICs measured by BACTEC 460TB and susceptibility testing at a lower pH of 5.5 confirmed genuine resistance. The pncA gene was sequenced in these four isolates and found not to have any mutations. This implies that an alternative mechanism of resistance exists in these strains. We conclude that genotypic assessment of pyrazinamide resistance is unreliable, because it depends on the identification of a single resistance mechanism. Phenotypic methods such as the BACTEC 460TB technique remain the best methods for pyrazinamide susceptibility testing. PMID- 11015385 TI - Rapid quantification and differentiation of human polyomavirus DNA in undiluted urine from patients after bone marrow transplantation. AB - A combined PCR assay was developed for the detection and typing of human polyomavirus (huPoV) in clinical samples, consisting of (i) a qualitative seminested PCR assay (snPCR) to discriminate between huPoV BK and JC and (ii) a high-throughput, quantitative TaqMan PCR assay (TM-PCR) for the general detection of huPoV. The TM-PCR detects huPoV DNA in a linear range from 10(7) to 10(1) copies per assay. In reproducibility runs, the inter- and intra-assay variabilities were < or =60 and < or =50%, respectively. The snPCR assay uses a set of four primers for the same region of the BK and JC viral genomes. In the first round of amplification, two general primers were used; in the second round, one of these general primers and two additional, BK- or JC-specific primers were used simultaneously to produce amplicons of different sizes specific for BK virus (246 bp) and JC virus (199 bp), respectively. We tested different urine dilutions in order to determine the inhibitory effects of urine on PCR amplification. Furthermore, we compared the use of native urine with DNA purified by different preparation procedures. Our results show, that a 1:10 dilution of the urine led to complete reduction of the amplification inhibition found with 6% of undiluted urine samples. In a clinical study including 600 urine specimens, our assay turned out to be fast, cheap, and reliable in both qualitative and quantitative aspects. PMID- 11015386 TI - Differentiation between Candida dubliniensis and Candida albicans by fatty acid methyl ester analysis using gas-liquid chromatography. AB - Candida dubliniensis is often found in mixed culture with C. albicans, but its recognition is hampered as the color of its colonies in primary culture on CHROMagar Candida varies. Furthermore, definite identification of C. dubliniensis is difficult to achieve, time-consuming, and expensive. Therefore, a method to discriminate between these two closely related yeast species by fatty acid methyl ester (FAME) analysis using gas-liquid chromatography (Sherlock Microbial Identification System [MIS]; MIDI, Inc., Newark, Del.) was developed. Although the chromatograms of these two species revealed no obvious differences when applying FAME analysis, a new library (CADLIB) was successfully created using Sherlock Library Generation Software (MIDI). The amount and frequency of FAME was analyzed using library training files (n = 10 for each species), preferentially those comprising reference strains. For testing the performance of the CADLIB, clinical isolates genetically assigned to the respective species (C. albicans, n = 32; C. dubliniensis, n = 28) were chromatographically analyzed. For each isolate tested, MIS computed a similarity index (SI) indicating a hierarchy of possible strain fits. When using the newly created library CADLIB, the SIs for C. albicans and C. dubliniensis ranged from 0.11 to 0.96 and 0.53 to 0. 93 (for all but one), respectively. Only three isolates of C. albicans (9.4%) were misidentified as C. dubliniensis, whereas all isolates of C. dubliniensis were correctly identified. Resulting differentiation accuracy was 90.6% for C. albicans and 100% for C. dubliniensis. Cluster analysis and principal component analysis of the resulting FAME profiles showed two clearly distinguishable clusters matching up with two assigned species for the strains tested. Thus, the created library proved to be well suited to discriminate between these two species. PMID- 11015387 TI - Expression of a gene encoding the major antigenic protein 2 homolog of ehrlichia chaffeensis and potential application for serodiagnosis. AB - The major antigenic protein 2 (MAP2) homolog of Ehrlichia chaffeensis was cloned and expressed. The recombinant protein was characterized and tested in an enzyme linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of human monocytic ehrlichiosis. The recombinant protein, which contained a C-terminal polyhistidine tag, had a molecular mass of approximately 26 kDa. The antigen was clearly identified by Western immunoblotting using antihistidine antibody. However, immune sera failed to react with the recombinant on immunoblots when the antigen was denatured by heat or reduced using beta mercaptoethanol. The recombinant MAP2 (rMAP2) was used in an ELISA format with 60 blinded serum samples. Twenty of the serum samples were previously demonstrated to contain antibodies reactive with E. chaffeensis by indirect immunofluorescence assays (IFAs). The remaining 40 samples were seronegative. All samples negative by IFA were also found to be negative for antibodies against the rMAP2 of E. chaffeensis by using the ELISA. Only 1 of 20 IFA-positive samples tested negative in the rMAP2 ELISA. There was 100% agreement using IFA-negative samples and 95% agreement using IFA-positive samples, resulting in a 97.5% overall agreement between the two assays. These data suggest that the rMAP2 homolog of E. chaffeensis may have potential as a test antigen for the serodiagnosis of human monocytic ehrlichiosis. To our knowledge, this recombinant is unique because it is thus far the only E. chaffeensis recombinant antigen that has been shown to work in an ELISA format. PMID- 11015388 TI - Two enzyme immunoassays and PCR for detection of Helicobacter pylori in stool specimens from pediatric patients before and after eradication therapy. AB - This study of pediatric patients was intended to determine the suitability of stool PCR and two antigen enzyme immunoassays (EIAs; Premier Platinum HpSA and the novel FemtoLab H. pylori), which detect Helicobacter pylori antigens in feces, as pretreatment diagnostic tools and especially as posttreatment control. Forty-nine H. pylori-infected children with dyspepsia received eradication therapy. Successful treatment was determined by a negative [(13)C]urea breath test 4 and 12 weeks after discontinuation of therapy. Fecal specimens were collected prior to eradication therapy as well as 4 weeks after the end of treatment. Successfully treated children delivered stool samples at 6, 8, and 12 weeks posttreatment also. Specimens were examined by seminested PCR and Premier Platinum HpSA and were reexamined by both EIAs as soon as FemtoLab H. pylori was available. In the first test series, the overall sensitivities of PCR and Premier Platinum HpSA were 93.0 and 91.1%, respectively. With specimens collected at 4 weeks after treatment, the respective specificities were 68.8 and 79.3%. After longer follow-up periods, however, they gradually increased to 100 and 96.9%, respectively. In the new test series, Premier Platinum HpSA delivered a considerably lower number of false-positive results (4 versus 18), indicating intertest variations. The overall test sensitivity was 94.6%, and the overall specificity was 97.5%. FemtoLab H. pylori showed an excellent performance with an overall sensitivity and specificity of 98.2 and 98.1%, respectively. Thus, in contrast to PCR, both EIAs were shown to be suitable for early posttreatment control. PMID- 11015389 TI - Evaluation of the Etest method for determining voriconazole susceptibilities of 312 clinical isolates of Candida species by using three different agar media. AB - Performance of the Etest for voriconazole susceptibility testing of 312 isolates of Candida spp. was assessed against that of the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose (RPG), Casitone agar (CAS), and antibiotic medium 3 (AM3) agar and were read after incubation for 48 h at 35 degrees C. The Candida spp. isolates included C. albicans (n = 174), C. glabrata (n = 55), C. tropicalis (n = 31), C. parapsilosis (n = 39), C. krusei (n = 5), C. lusitaniae (n = 2), and C. guilliermondii (n = 6). The Etest results obtained using RPG correlated well with the reference MICs. Overall agreement ranged from 91% for C. glabrata to 100% for C. tropicalis, C. parapsilosis, C. guilliermondii, C. krusei, and C. lusitaniae. When CAS was used, agreement ranged from 80% for C. krusei to 100% for C. parapsilosis, C. guilliermondii, and C. lusitaniae. With AM3, agreement ranged from 58% for C. glabrata to 100% for C. lusitaniae and C. guilliermondii. The Etest method using RPG appears to be a useful method for determining voriconazole susceptibilities of Candida species. PMID- 11015390 TI - Immunodiagnosis of Echinococcus infections: confirmatory testing and species differentiation by a new commercial Western Blot. AB - The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID- 11015391 TI - Helicobacter aurati sp. nov., a urease-positive Helicobacter species cultured from gastrointestinal tissues of Syrian hamsters. AB - A novel helicobacter with the proposed name Helicobacter aurati (type strain MIT 97-5075c) has been isolated from the inflamed stomachs and ceca of adult Syrian hamsters. The new species is fusiform with multiple bipolar sheathed flagella and periplasmic fibers; it contains urease and gamma-glutamyl transpeptidase. By 16S rRNA sequencing and repetitive element PCR-based DNA fingerprinting, it was found that H. aurati represents a distinct taxon and clusters with Helicobacter muridarum, Helicobacter hepaticus, and Helicobacter sp. MIT 94-022. H. aurati was recovered from hamsters housed in various research and vendor facilities. Further studies are necessary to define its association with disease and other microbiota in hamsters, as well as its impact on research projects involving hamsters. H. aurati (GenBank accession number AF297868) can be used in animal experiments to define the factors that are important for gastric helicobacter pathogenesis. PMID- 11015392 TI - Molecular and antigenic characterization of a highly evolved derivative of the type 2 oral poliovaccine strain isolated from sewage in Israel. AB - An unusual, highly diverged derivative of the Sabin type 2 oral poliovaccine (OPV) strain was recovered from environmental samples during routine screening for wild polioviruses. Virus was cultivated in L20B cells and then passaged on BGM cells at 40 degrees C (RCT [reproductive capacity at supraoptimal temperature]-positive marker) to select against most OPV strains. All but 1 of 25 RCT-positive OPV-derived environmental isolates were antigenically and genetically (>99.5% VP1 sequence match) similar to the respective Sabin strains. However, isolate PV2/4568-1/ISR98 (referred to below as 4568-1) escaped neutralization with Sabin 2-specific monoclonal antibodies and cross-adsorbed sera, and had multiple nucleotide substitutions (220 of 2,646; 8.3%) in the P1 capsid region. Fourteen of the 44 associated amino acid substitutions in the capsid mapped to neutralizing antigenic sites. Neutralizing titers in the sera of 50 Israeli children 15 years old were significantly lower to 4568-1 (geometric mean titer [GMT], 47) than to Sabin 2 (GMT, 162) or to the prototype wild strain, PV2/MEF-1/EGY42 (GMT, 108). Two key attenuating sites had also reverted in 4568-1 (A(481) to G in the 5' untranslated region and the VP1 amino acid I(143) to T), and the isolate was highly neurovirulent for transgenic mice expressing the poliovirus receptor (PVR-Tg21 mice). The extensive genetic divergence of 4568-1 from the parental Sabin 2 strain suggested that the virus had replicated in one or more people for approximately 6 years. The presence in the environment of a highly evolved, neurovirulent OPV-derived poliovirus in the absence of polio cases has important implications for strategies for the cessation of immunization with OPV following global polio eradication. PMID- 11015393 TI - Development of a PCR-based line probe assay for identification of fungal pathogens. AB - We report on a reverse-hybridization line probe assay (LiPA) which when combined with PCR amplification detects and identifies clinically significant fungal pathogens including Candida, Aspergillus, and Cryptococcus species. DNA probes have been designed from the internal transcribed-spacer (ITS) regions of Candida albicans, Candida parapsilosis, Candida glabrata, Candida tropicalis, Candida krusei, Candida dubliniensis, Cryptococcus neoformans, Aspergillus fumigatus, Aspergillus versicolor, Aspergillus nidulans and Aspergillus flavus. The probes were incorporated into a LiPA for detection of biotinylated ITS PCR products, and the specificity of the probes was evaluated. We established LiPA detection limits for ITS 1 and for full ITS amplicons for genomic DNA from C. albicans, A. fumigatus, and C. neoformans. Further evaluation of the LiPA was carried out on clinical fungal isolates. One hundred twenty-seven isolates consisting of dimorphic yeasts and dermatophytic and filamentous fungi were tested by the LiPA, which correctly identified 77 dimorphic yeasts and 23 of the filamentous isolates; the remaining 27 isolates represented species of fungi for which probes were not included in the LiPA. The fungal-PCR-LiPA technology was applied to blood samples inoculated with Candida cells which were pretreated by minibead beating to mechanically disrupt the cells, with the DNA extracted by either a previously described guanidium thiocyanate-silica method or the commercially available QIAmp tissue kit. PCR amplification of the extracted DNA and subsequent DNA probe hybridization in the LiPA assay yielded detection limits of 2 to 10 cells/ml. An internal standard control was included in the PCR amplification to monitor for PCR inhibition. This fungal PCR-LiPA assay is robust and sensitive and can easily be integrated into a clinical-testing laboratory with the potential for same-day diagnosis of fungal infection. PMID- 11015394 TI - Comparison of the CMV brite turbo assay and the digene hybrid capture CMV DNA (Version 2.0) assay for quantitation of cytomegalovirus in renal transplant recipients. AB - We compared the CMV Brite Turbo Kit (BT) and the Digene Hybrid Capture CMV DNA (version 2.0) assay (HC2) in the quantitation of pp65 antigenemia and cytomegalovirus (CMV) DNA levels in immunosuppressed renal transplant recipients. Of 123 blood specimens collected from 24 renal transplant recipients, BT and HC2 assays detected 35 and 39 positive samples, respectively. The overall concordance rate between the two assays was 90%. Discordant results were observed at low levels of viremia, so that 8 samples were HC2 positive but BT negative and another 4 were BT positive but HC2 negative. There was good correlation (R(2) = 0.766; P<0.01) between the levels of CMV DNA and pp65 antigenemia in the 31 concordant positive samples. Correlation between results obtained with the two assays was confirmed by longitudinal studies for a patient who developed clinical CMV disease. HC2 may be more sensitive at low viremia levels and allow earlier detection of impending CMV disease. The BT assay offered the advantage of a rapid (2-h) turnaround time. We conclude that BT and HC2 assays have similar sensitivity and efficacy in the diagnosis and monitoring of CMV infection and disease in renal transplant recipients. While the HC2 assay would be appropriate for centers that handle a large number of samples, the BT test may be more suitable for small sample numbers or when results are needed urgently. PMID- 11015395 TI - Characterization of Staphylococcus aureus coagulase type VII isolates from staphylococcal food poisoning outbreaks (1980-1995) in Tokyo, Japan, by pulsed field gel electrophoresis. AB - Staphylococcus aureus coagulase type VII strains have been the strains most frequently isolated from staphylococcal food poisoning outbreaks in Tokyo, Japan. We applied pulsed-field gel electrophoresis (PFGE) of chromosomal DNA digested with SmaI to characterize 129 coagulase type VII strains. These were isolated from 129 cases occurring in outbreaks in 35 districts during a 16-year period (1980-1995). The 129 outbreak strains were classified into three types, designated A (n = 115), B (n = 10), and C (n = 4). Types A and C were further divided into 33 (A1 to A33) and 4 (C1 to C4) subtypes, respectively. Strains of the same subtypes were isolated from food poisoning cases in the same districts at time intervals of 1 or 2 to 5 years. PFGE typing appears to be a useful method for subdividing strains of S. aureus coagulase type VII. A combination of coagulase typing and PFGE typing would provide more detailed information than the former method alone in epidemiologic investigations of staphylococcal food poisoning. PMID- 11015396 TI - Utilization of exocellular mannan from Rhodotorula glutinis as an immunoreactive antigen in diagnosis of leptospirosis. AB - Previously, Rhodotorula glutinis was reported to produce a large amount of exocellular mannan, having a repeating unit of -->3)-D-Manp-(1-->4)-D-Manp-(1-->. Recently, we found that antigenic polysaccharides of Leptospira biflexa serovar patoc strain Patoc I have the same repeating unit and cross-react with antisera raised against extended strains of other leptospires (K. Matsuo, E. Isogai, and Y. Araki, Carbohydr. Res., in press). This structural identity and the difficulty of producing and isolating antigens led us to confirm the usefulness of Rhodotorula mannan as an immunoreactive antigen in a serological diagnosis of leptospirosis. In the present investigation, we confirmed the structural identity of an exocellular mannan isolated from R. glutinis AHU 3479 and tried to use it as an immunoreactive antigen in a serological diagnosis of leptospirosis. From its chemical analysis and (1)H- and (13)C-labeled nuclear magnetic resonance spectrometry, the Rhodotorula mannan was confirmed to consist of the same disaccharide units. Furthermore, such a preparation was shown to immunoreact to various sera from patients suffering with leptospirosis as well as to most rabbit antiserum preparations obtained from immunization with various strains of pathogenic leptospires. Therefore, the Rhodotorula mannan preparation is useful as an immunoreactive antigen in the serological diagnosis for leptospirosis. PMID- 11015397 TI - Evaluation of a PCR primer based on the isocitrate dehydrogenase gene for detection of Helicobacter pylori in feces. AB - In order to improve detection and identification of Helicobacter pylori in highly contaminated samples, we evaluated new specific primers based on the DNA base sequence within the isocitrate dehydrogenase (icd) gene to amplify a 1,200-bp DNA segment. The specificity of the icd primer was tested against DNA derived from various bacteria, including 7 Helicobacter species and a panel of 1 gram variable, 2 gram-positive, and 16 gram-negative bacteria, as well as DNA from houseflies and feces from H. pylori-negative patients. The primers permitted the detection of all clinical H. pylori isolates tested, but no reactions were observed with negative controls. Several procedures for DNA extraction from feces were evaluated using PCR with icd primers. The lower limits of detection of H. pylori DNA from two different sources containing the same number of H. pylori organisms, a pure culture and feces spiked with H. pylori, were established for each extraction method tested. The results were 8.0 x 10(3) CFU/ml for cultures of pure H. pylori, and 8.0 x 10(6) CFU/ml for H. pylori from feces, using the phenol-chloroform method; 8.0 x 10(2) and 7.0 x 10(3) CFU/ml, respectively, for a glass matrix and chaotropic solution protocol; 8.0 x 10(2) and 7.0 x 10(3) CFU/ml, respectively, for the QIAamp tissue kit; and 5.0 x 10(2) and 5.0 x 10(3) CFU/ml, respectively, for the XTRAX DNA extraction kit. We conclude that the use of the icd gene as a primer for PCR represents a specific and sensitive assay for detection of H. pylori in highly contaminated samples. PMID- 11015398 TI - Prevalence of O1/K1- and O2/K3-reactive Actinobacillus suis in healthy and diseased swine. AB - A cell surface antigen-typing system was devised for the swine pathogen Actinobacillus suis and used to examine the prevalence of different lipopolysaccharide (O) types in healthy and diseased pigs. The strains examined in this study were isolated from a variety of locations in Canada and from Kansas. Lipopolysaccharide preparations of 151 isolates of A. suis were characterized by immunoblotting using polyclonal antisera generated to strains SO4 (O1/K1), H89-1173 (O2/K3), and VSB 3714, a rough strain. Approximately 54% (62 of 114) of A. suis isolates from diseased pigs, all (11 of 11) isolates from healthy pigs, and all (4 of 4) reference strains reacted with O1/K1 antiserum. More than 80% (18 of 22) of A. suis strains used for bacterin production and approximately 41% (47 of 114) of isolates from diseased pigs bound O2/K3 antiserum. One isolate appeared to be rough, and five were untypeable. O1/K1- and O2/K3-reactive strains were equally prevalent in Kansas, whereas O2/K3-reactive strains were more common in Quebec and western Canada and O1/K1 strains were most common in Ontario. The fact that virtually all of the strains submitted for bacterin production were O2/K3-reactive strains is consistent with the notion that these strains may be more virulent than O1/K1 strains; alternatively, this may reflect geographic or other biases. In addition, we observed cross-reactivity between A. suis cell surface antigens and swine antisera to several other important pathogens. This finding may explain why previous attempts to develop a simple serodiagnostic test for A. suis have been unsuccessful. PMID- 11015400 TI - Diagnosis of cutaneous leishmaniasis in Colombia: the sampling site within lesions influences the sensitivity of parasitologic diagnosis. AB - Parasitologic confirmation of cutaneous leishmaniasis is obligatory before chemotherapy can be considered. Direct microscopic examination of scrapings taken from indurated borders of ulcers has been routinely used as primary method of diagnosis. In this report we compared the sensitivity of examination of dermal scrapings taken from the bottoms of ulcers (BDS) with that of dermal scrapings taken from indurated active margins of lesions (MDS) in a total of 115 patients. The sensitivities of the microscopic examination were 90.4 and 78.3% for BDS and MDS samples, respectively. When the PCR method was used with a group of 40 patients, we also observed a higher sensitivity when BDS samples were examined (80.8% in BDS samples versus 57.7% in MDS samples). The improvement of the diagnostic sensitivity in the BDS samples appears to be related to the higher parasite load and more easily detectable morphology of amastigotes in the centers of the ulcers. Other parasitologic diagnostic methods, such as culture and histopathologic examination of biopsies, are less sensitive (67.5 and 64.3%, respectively). Aspirate culture, however, was shown to be the most sensitive method for the diagnosis of patients with chronic ulcers. When microscopic examinations of both MDS and BDS samples are combined, the sensitivity of diagnosis may rise up to 94%. We therefore recommend this method as a primary routine procedure for diagnosis of cutaneous leishmaniasis. PMID- 11015399 TI - PCR assessment of Chlamydia trachomatis infection of semen specimens processed for artificial insemination. AB - In order to ascertain the microbiological quality of stored semen specimens processed for artificial insemination by a donor (AID), we developed a PCR assay targeting the chlamydial plasmid to detect Chlamydia trachomatis in semen. The lower limit of detection of this assay corresponded to 2.5 to 5 elementary bodies per microl of semen. A total of 669 cryopreserved ejaculates from 97 asymptomatic donors were tested for C. trachomatis infection. Twelve ejaculates, originating from four donors, were found to be positive, indicating a 4% prevalence of C. trachomatis infection among the donor population studied. Cross-contamination between the cryopreserved specimens in the storage container was studied by typing using sequence analysis of PCR-amplified omp1 genes of the strains. Two donors were infected with serovar E, one was infected with serovar F, and one was infected with serovar K. For two donors, the duration of C. trachomatis positivity could be assessed. One donor donated C. trachomatis-positive semen for at least 4 successive months, and the other did so for at least 16 months. The occurrence of C. trachomatis infection in cryopreserved donor semen indicates that ejaculates from donors not tested for a C. trachomatis infection just prior to donation should be tested for infection by a direct test such as the PCR described here. Direct testing of semen specimens will detect not only donors with an active infection but also C. trachomatis-infected ejaculates already stored and will thus improve the microbiological quality of AID, since discrepancies in the presence of C. trachomatis in urine and semen specimens have been reported. PMID- 11015402 TI - Bacillus anthracis diversity in Kruger National Park. AB - The Kruger National Park (KNP), South Africa, has a recorded history of periodic anthrax epidemics causing widespread disease among wild animals. Bacillus anthracis is the causative agent of anthrax, a disease primarily affecting ungulate herbivores. Worldwide there is little diversity among B. anthracis isolates, but examination of variable-number tandem repeat (VNTR) loci has identified six major clones, with the most dissimilar types split into the A and B branches. Both the A and B types are found in southern Africa, giving this region the greatest genetic diversity of B. anthracis worldwide. Consequently, southern Africa has been hypothesized to be the geographic origin of B. anthracis. In this study, we identify the genotypic types of 98 KNP B. anthracis isolates using multiple-locus VNTR analysis. Two major types are evident, the A branch and the B branch. The spatial and temporal distribution of the different genotypes indicates that anthrax epidemic foci are independent, though correlated through environmental cues. Kruger B isolates were found on significantly higher calcium and higher-pH soils than were Kruger type A. This relationship between genotype and soil chemistry may be due to adaptive differences among divergent anthrax strains. While this association may be simply fortuitous, adaptation of A types to diverse environmental conditions is consistent with their greater geographic dispersal and genetic dissimilarity. PMID- 11015403 TI - Prevalence of enterotoxin genes in Aeromonas spp. isolated from children with diarrhea, healthy controls, and the environment. AB - Aeromonads are causative agents of a number of human infections. Even though aeromonads have been isolated from patients suffering from diarrhea, their etiological role in gastroenteritis is unclear. In spite of a number of virulence factors produced by Aeromonas species, their association with diarrhea has not been clearly linked. Recently, we have characterized a heat-labile cytotonic enterotoxin (Alt), a heat-stable cytotonic enterotoxin (Ast), and a cytotoxic enterotoxin (Act) from a diarrheal isolate of Aeromonas hydrophila. Alt and Ast are novel enterotoxins which are not related to cholera toxin; Act is aerolysin related and has hemolytic, cytotoxic, and enterotoxic activities. We studied the distribution of the alt, ast, and act enterotoxin genes in 115 of 125 aeromonads isolated from 1, 735 children with diarrhea, in all 27 aeromonads isolated from 830 control children (P = 7 x 10(-4) for comparison of rates of isolation of aeromonads from cases versus those from controls), and in 120 randomly selected aeromonads from different components of surface water in Bangladesh. Aeromonas isolates which were positive only for the presence of the alt gene had similar distributions in the three sources; the number of isolates positive only for the presence of the ast gene was significantly higher for the environmental samples than for samples from diarrheal children; and isolates positive only for the presence of the act gene were not found in any of the three sources. Importantly, the number of isolates positive for both the alt and ast genes was significantly higher for diarrheal children than for control children and the environment. Thus, this is the first study to indicate that the products of both the alt and ast genes may synergistically act to induce severe diarrhea. In 26 patients, Aeromonas spp. were isolated as the sole enteropathogen. Analysis of clinical data from 11 of these patients suggested that isolates positive for both the alt and ast genes were associated with watery diarrhea but that isolates positive only for the alt gene were associated with loose stools. Most of the isolates from the three sources could be classified into seven phenospecies and eight hybridization groups. For the first time, Aeromonas eucrenophila was isolated from two children, one with diarrhea and another without diarrhea. PMID- 11015404 TI - Phenotypic and genetic characterization of Lactococcus garvieae isolated in Spain from lactococcosis outbreaks and comparison with isolates of other countries and sources. AB - The phenotypic and genetic analysis results for 84 isolates of Lactococcus garvieae (including 62 strains from trout with lactococcosis from four different countries, 7 strains from cows and water buffalos with subclinical mastitis, 3 from water, and 10 from human clinical samples) are presented. There was great phenotypic heterogeneity (13 different biotypes) based on the acidification of saccharose, tagatose, mannitol, and cyclodextrin and the presence of the enzymes pyroglutamic acid arylamidase and N-acetyl-beta-glucosaminidase. L. garvieae also exhibited high genetic diversity by pulsed-field gel electrophoresis (PFGE), with 19 different pulsotypes among the isolates of L. garvieae studied. Only epidemiologically related strains, like the Spanish and Italian fish isolates and the cow and water buffalo isolates, displayed a close genetic relationship by PFGE, while the strains isolated from sporadic clinical cases, like the human isolates, were genetically unrelated. Overall, a general correlation between phenotypic and genetic data was observed. Epidemiological analysis of biotype and PFGE results indicated that the trout lactococcosis outbreaks in Spain and Portugal and those in France and Italy were produced by genetically unrelated clones. In Spain, two different clones were detected; the outbreaks diagnosed from 1995 onward were produced by a clone (biotype 2, pulsotype A1) which, although genetically related, was different from the one that was responsible for the outbreaks studied between 1991 and 1994 (biotype 1, pulsotype B). The Portuguese isolate had a biochemical profile identical to that of the Spanish strain isolated from 1995 onward and is also genetically closely related to this strain (pulsotype A2). There was a close relationship between the two pulsotypes (E and F) found in the Italian isolates. The French isolate (biotype 3, pulsotype D) was not genetically related to any other L. garvieae fish isolate. These results suggest the existence of diverse infection sources for the different lactococcosis outbreaks. PMID- 11015401 TI - Class 1 integron-borne, multiple-antibiotic resistance encoded by a 150-kilobase conjugative plasmid in epidemic vibrio cholerae O1 strains isolated in Guinea Bissau. AB - In the 1996-1997 cholera epidemic in Guinea-Bissau, surveillance for antimicrobial resistance showed the emergence of a multidrug-resistant strain of Vibrio cholerae O1 during the course of the epidemic. The strain was resistant to ampicillin, erythromycin, tetracycline, furazolidone, aminoglycosides, trimethoprim, and sulfamethoxazole. Concomitant with the emergence of this strain, we observed a resurgence in the number of registered cholera cases as well as an increase in the case fatality rate from 1.0% before the emergence of the multiple-drug-resistant strain to 5.3% after the emergence of the strain. Our study shows that the strain contained a 150-kb conjugative multiple-antibiotic resistance plasmid with class 1 integron-borne gene cassettes encoding resistance to trimethoprim (dhfrXII) and aminoglycosides [ant(3")-1a]). The finding of transferable resistance to almost all of the antibiotics commonly used to treat cholera is of great public health concern. Studies should be carried out to determine to what extent the strain or its resistance genes have been spread to other areas where cholera is endemic. PMID- 11015405 TI - Rapid and sensitive plate method for detection of Aspergillus fumigatus. AB - The routine identification of Aspergillus fumigatus in clinical samples involves, apart from direct examination, the isolation of the organism on a plate followed by its microscopic characterization. This approach lacks sensitivity, specificity, and speed. A new procedure has been developed combining microcolony formation on a nylon membrane filter at 45 degrees C with the detection of a specific 4-methylumbelliferyl-alpha-L-arabinopyranoside cleaving enzyme activity in digitonin permeabilized cells. The test takes approximately 14 h and has an efficiency of 98.2% and false-positive and -negative rates of 0 and 3.1%, respectively. When applied to 188 clinical samples taken from patients with proven or nonproven presence of Aspergillus species, a good agreement with the conventional plate-microscopy method was obtained. PMID- 11015406 TI - Evaluation of phenotypic and genotypic methods for subtyping Campylobacter jejuni isolates from humans, poultry, and cattle. AB - Six methods for subtyping of Campylobacter jejuni were compared and evaluated with a collection of 90 isolates from poultry, cattle, and sporadic human clinical cases as well as from a waterborne outbreak. The applied methods were Penner heat-stable serotyping; automated ribotyping (RiboPrinting); random amplified polymorphic DNA typing (RAPD); pulsed-field gel electrophoresis (PFGE); restriction fragment length polymorphisms of the flagellin gene, flaA (fla-RFLP); and denaturing gradient gel electrophoresis of flaA (fla-DGGE). The methods were evaluated and compared on the basis of their abilities to identify isolates from one outbreak and discriminate between unrelated isolates and the agreement between methods in identifying clonal lines. All methods identified the outbreak strain. For a collection of 80 supposedly unrelated isolates, RAPD and PFGE were the most discriminatory methods, followed by fla-RFLP and RiboPrinting. fla-DGGE and serotyping were the least discriminative. All isolates included in this study were found to be typeable by each of the methods. Thirteen groups of potentially related isolates could be identified using a criterion that at least four of the methods agreed on clustering of isolates. None of the subtypes could be related to only one source; rather, these groups represented isolates from different sources. Furthermore, in two cases isolates from cattle and human patients were found to be identical according to all six methods. PMID- 11015407 TI - Discrimination between viable and dead Encephalitozoon cuniculi (Microsporidian) spores by dual staining with sytox green and calcofluor white M2R. AB - Microsporidia are obligate intracellular parasites, recognized as causing chronic diarrhea and systemic disease in AIDS patients, organ transplant recipients, travelers, and malnourished children. Species of microsporidia that infect humans have been detected in drinking-water sources, and methods are needed to ascertain if these microsporidia are viable and capable of causing infections. In this study, Calcofluor White M2R and Sytox Green stains were used in combination to differentiate between live (freshly harvested) and dead (boiled) Encephalitozoon cuniculi spores. Calcofluor White M2R binds to chitin in the microsporidian spore wall. Dual-stained live spores appeared as turquoise-blue ovals, while dead spores appeared as white-yellow ovals at an excitation wavelength of 395 to 415 nm used for viewing the Calcofluor stain. Sytox Green, a nuclear stain, is excluded by live spores but penetrates compromised spore membranes. Dual-stained dead spores fluoresced bright yellow-green when viewed at an excitation wavelength of 470 to 490 nm, whereas live spores failed to stain with Sytox Green. After live and dead spores were mixed at various ratios, the number of viably stained spores detected in the dual-staining procedure correlated (P = 0.0025) with the expected numbers of viable spores. Spore mixtures were also assayed for infectivity in a focus-forming assay, and a correlation (P = 0.0002) was measured between the percentage of focus-forming microsporidia and the percentage of expected infectious spores in each mixture. By analysis of variance, no statistically significant differences were measured between the percentage of viably stained microsporidia and the percentage of infectious microsporidia (P = 0.964) in each mixture. These results suggest that Calcofluor White M2R and Sytox Green stains, when used together, may facilitate studies to identify viable microsporidia. PMID- 11015408 TI - Molecular epidemiology of Entamoeba spp.: evidence of a bottleneck (Demographic sweep) and transcontinental spread of diploid parasites. AB - Entamoeba histolytica causes amebic colitis and liver abscess in developing countries such as Mexico and India. Entamoeba dispar is morphologically identical but is not associated with disease. Here we determined the ploidy of E. histolytica and developed PCR-based methods for distinguishing field isolates of E. histolytica or E. dispar. Fluorescence in situ hybridization showed that E. histolytica trophozoites are diploid for five "single-copy" probes tested. Intergenic sequences between superoxide dismutase and actin 3 genes of clinical isolates of E. histolytica from the New and Old Worlds were identical, as were those of E. dispar. These results suggest a bottleneck or demographic sweep in entamoebae which infect humans. In contrast, E. histolytica and E. dispar genes encoding repeat antigens on the surface of trophozoites (Ser-rich protein) or encysting parasites (chitinase) were highly polymorphic. chitinase alleles suggested that the early axenized strains of E. histolytica, HM-1 from Mexico City, Mexico, and NIH-200 from Calcutta, India, are still present and that similar E. dispar parasites can be identified in both the New and Old Worlds. Ser rich protein alleles, which suggested the presence of the HM-1 strain in Mexico City, included some E. histolytica genes that predicted Ser-rich proteins with very few repeats. These results, which suggest diversifying selection at chitinase and Ser-rich protein loci, demonstrate the usefulness of these alleles for distinguishing clinical isolates of E. histolytica and E. dispar. PMID- 11015409 TI - Detection and quantitation of human immunodeficiency virus type 1 in the female genital tract. The Division of AIDS Treatment Research Initiative 009 Study Group. AB - Human immunodeficiency virus type 1 (HIV-1) was detected in the genital tracts of 59% of 225 women by RNA PCR and in 7% of the women by culture. In a comparison of two sampling methods, endocervical swabs were more sensitive than cervicovaginal lavage for HIV-1 RNA detection by PCR but not by culture and their sensitivity was independent of the concentration of HIV-1 RNA. PMID- 11015410 TI - InTray GC medium versus modified Thayer-Martin agar plates for diagnosis of gonorrhea from endocervical specimens. AB - A new self-contained medium system for the cultivation of Neisseria gonorrhoeae was compared to modified Thayer-Martin medium for the diagnosis of gonorrhea from endocervical specimens. There was no difference in the ability of the two methods to support the growth of N. gonorrhoeae. PMID- 11015412 TI - Comparison between plasma and whole blood specimens for detection of Aspergillus DNA by PCR. AB - Ninety-six plasma and whole blood specimens from nine selected patients were analyzed for the presence of Aspergillus DNA. Nineteen specimens from three patients with proven aspergillosis were PCR positive in both materials, whereas an additional 22 were PCR positive in whole blood only. All 36 samples from six patients without signs of aspergillosis were negative in both assays. We conclude that although plasma and whole blood spiked with Aspergillus conidia showed an identical lower detection limit (10 CFU), the sensitivity of plasma PCR was lower than that of PCR performed on whole blood samples. PMID- 11015411 TI - Performance of five agar media for recovery of fungi from isolator blood cultures. AB - We studied the recovery of 1,270 fungal isolates from 176,144 Isolator blood cultures (0.72% positive) on bacterial and fungal media, under routine and differing incubation conditions. Except with Histoplasma capsulatum, chocolate agar incubated for only 3 days proved to be an excellent medium for the recovery of fungi from the Isolator system. PMID- 11015413 TI - Evaluation of Etest for susceptibility testing of Mycobacterium tuberculosis. AB - The Etest method for susceptibility testing of Mycobacterium tuberculosis was compared to the agar proportion method using four first-line agents and two fluoroquinolones. Catergorical agreement between the methods was 100% for rifampin, ethambutol, streptomycin, and ofloxacin and 98% for isoniazid. Results were obtained in 6 to 10 days by Etest. The Etest method is suitable for testing the agents evaluated against M. tuberculosis. PMID- 11015414 TI - Distribution of a specific 500-base-pair fragment in mycobacterium bovis isolates from Sardinian cattle. AB - Amplification of a specific, 500-bp fragment from Mycobacterium bovis isolates and use of the fragment to differentiate between Mycobacterium tuberculosis and M. bovis was previously reported (J. G. Rodriguez, G. A. Meja, P. Del Portillo, M. E. Patarroyo, and L. A. Murillo, Microbiology 141:2131-2138, 1995). In the present study, 30 M. bovis isolates from Sardinian cattle were examined for the presence of this 500-bp fragment; 4 of the 30 isolates lacked the fragment. This result indicates that identification of M. bovis strains by amplification of the 500-bp sequence may lead to false-negative results. PMID- 11015415 TI - Utility of tissue culture for detection of Toxoplasma gondii in vitreous humor of patients diagnosed with toxoplasmic retinochoroiditis. AB - Laboratory recovery and confirmation of the etiologic agent in necrotizing retinochoroiditis are problematic. Tissue culture and intraocular antibody titers were compared as adjuncts to clinical diagnosis for toxoplasmic retinochoroiditis: the correlations were 91 and 67%, respectively. Isolation of Toxoplasma gondii may establish a definitive diagnosis in patients with toxoplasmic retinochoroiditis. PMID- 11015416 TI - Use of amplified fragment length polymorphisms for typing Corynebacterium diphtheriae. AB - Amplified fragment length polymorphism (AFLP) was investigated for the differentiation of Corynebacterium diphtheriae isolates. Analysis using Taxotron revealed 10 distinct AFLP profiles among 57 isolates. Strains with ribotype patterns D1, D4, and D12 could not be distinguished; however, the technique discriminated isolates of ribotype patterns D3, D6, and D7 further. AFLP was rapid, fairly inexpensive, and reproducible and could be used as an alternative to ribotyping. PMID- 11015418 TI - G protein variation in respiratory syncytial virus group A does not correlate with clinical severity. AB - Respiratory syncytial virus group A strain variations of 28 isolates from The Netherlands collected during three consecutive seasons were studied by analyzing G protein sequences. Several lineages circulated repeatedly and simultaneously during the respective seasons. No relationships were found between lineages on the one hand and clinical severity or age on the other. PMID- 11015417 TI - A probable new Helicobacter species isolated from a patient with bacteremia. AB - A probable new Helicobacter species was isolated from the blood of a 14-month-old aboriginal child who presented with vomiting, diarrhea, fever, and dry cough. The most similar 16S rRNA gene sequence was that of Helicobacter fennelliae CCUG 18820(T) but the new sequence differed from it by at least 32 base substitutions and by the presence of a large (353-nucleotide) intervening sequence. PMID- 11015419 TI - Evaluation of the Alexon-trend ProSpecT Campylobacter microplate assay. AB - We evaluated stool specimens known to contain or be free of Campylobacter by traditional culture, using the ProSpecT Campylobacter microplate assay (Alexon Trend, Ramsey, Minn.). This rapid enzyme immunoassay for the detection of Campylobacter-specific antigens demonstrated 96% sensitivity and 99% specificity and is an acceptable alternative method of Campylobacter detection. PMID- 11015420 TI - Absence of serotype-specific surface antigen and altered teichoic acid glycosylation among epidemic-associated strains of Listeria monocytogenes. AB - Outbreaks of food-borne listeriosis have often involved strains of serotype 4b. Examination of multiple isolates from three different outbreaks revealed that ca. 11 to 29% of each epidemic population consisted of strains which were negative with the serotype-specific monoclonal antibody c74.22, lacked galactose from the teichoic acid of the cell wall, and were resistant to the serotype 4b-specific phage 2671. PMID- 11015421 TI - Comparison of six commercial DNA extraction kits for recovery of cytomegalovirus DNA from spiked human specimens. AB - We evaluated six commercially available DNA extraction kits for their ability to recover DNA from various dilutions of cytomegalovirus (CMV) added to four different specimens: bronchoalveolar lavage, cerebral spinal fluid, plasma, and whole blood. The kits evaluated included the Puregene DNA isolation kit (PG), Generation Capture Column kit, MasterPure DNA purification kit, IsoQuick nucleic acid extraction kit, QIAamp blood kit, and NucliSens isolation kit (NS). All six kits evaluated effectively removed PCR inhibitors from each of the four specimen types and produced consistently positive results down to a spiked concentration of 200 PFU of whole CMV per ml. However, the NS and PG resulted in the most consistently positive results at the lowest concentrations of spiked CMV (4 and 0.4 PFU/ml) and, in this evaluation, offered the most sensitive methods for extracting CMV DNA from the four different spiked specimens. Processing time and cost were also evaluated. PMID- 11015422 TI - Clinical comparison of nonvented aerobic BacT/Alert blood culture bottle and standard aerobic bottle for detection of microorganisms in blood. AB - The current BacT/Alert standard aerobic (VA) blood culture bottle was redesigned and designated a nonvented aerobic (NVA) culture bottle; this bottle does not require venting. A total of 3,873 sets of blood samples for culture were obtained from adult patients with suspected bacteremia or fungemia. The NVA bottle showed performance equivalent to that of the VA bottle for recovery and speed of detection of microorganisms from blood without the need for venting the bottle. PMID- 11015423 TI - Molecular analysis of methicillin-resistant Staphylococcus aureus as a causative agent of bronchopulmonary infection: relation to colonization in the upper respiratory tract. AB - Using five diagnostic markers, we compared the types of 72 strains of methicillin resistant Staphylococcus aureus (MRSA) isolated simultaneously from the nasal cavity, pharynx, and sputum from 24 patients. Almost identical MRSA types had colonized the nasal cavity and sputum from the same patient for 21 (88%) of the patients. We speculate that most MRSA organisms isolated in sputum are derived from the nasal cavity, while a few are derived from the pharynx. PMID- 11015424 TI - Isolation of Moraxella canis from an ulcerated metastatic lymph node. AB - Moraxella canis was isolated in large numbers from an ulcerated supraclavicular lymph node of a terminal patient, who died a few days later. Although the patient presented with septic symptoms and with a heavy growth of gram-negative diplococci in the lymph node, blood cultures remained negative. M. canis is an upper-airway commensal from dogs and cats and is considered nonpathogenic for humans, although this is the third reported human isolate of this species. PMID- 11015425 TI - Evaluation of different preservation and storage methods for Malassezia spp. AB - Freezing at -80 degrees C, lyophilization, preservation in distilled water, and storage in different culture media were performed in order to find a suitable method that allowed a prolonged storage of Malassezia spp. Freezing at -80 degrees C was the only method successful at maintaining all species. PMID- 11015427 TI - Vancomycin-intermediate Staphylococcus aureus in Korea. AB - Recent reports on some methicillin-resistant Staphylococcus aureus (MRSA) with reduced susceptibility to vancomycin have been a major concern in Korea because of the widespread use of vancomycin due to a high prevalence of MRSA in the country. We describe a 45-year-old man with long-standing pelvic abscess due to MRSA. In spite of vancomycin and teicoplanin treatment for a long period of time, the patient died from MRSA sepsis. The blood culture isolate of MRSA exhibited reduced susceptibility to vancomycin (MIC, 8 microg/ml). This is the first report of a vancomycin-intermediate S. aureus case from Korea. PMID- 11015426 TI - VP4 and VP7 genotyping by reverse transcription-PCR of human rotavirus in mexican children with acute diarrhea. AB - Dual typing (VP4 and VP7) of rotavirus obtained from 257 Mexican children during three epidemiological seasons was performed by reverse transcription-PCR. The P1G1 genotype was the most prevalent (40%), followed by P1G3 (19%) and P2G2 (16%). Thirty-one specimens (12%) presented mixed infections, while some genotypes were not found. This is the first dual typing of isolates from diarrhea cases in Mexico. PMID- 11015429 TI - Osteomyelitis caused by Staphylococcus schleiferi and evidence of misidentification of this Staphylococcus species by an automated bacterial identification system. AB - We report a case of sternal osteomyelitis due to Staphylococcus schleiferi in a patient who underwent thoracic surgery. This constitutes the first documented case of osteomyelitis caused by this Staphylococcus species. We also relate our experience in the utilization of commercially available MicroScan panels for the identification of this microorganism. PMID- 11015428 TI - Evaluation of the abbott LCx HIV-1 RNA quantitative, a new assay for quantitative determination of human immunodeficiency virus type 1 RNA. AB - A new quantitative reverse transcription (RT)-PCR assay for human immunodeficiency virus type 1 (HIV-1) RNA (Abbott LCx HIV RNA Quantitative assay) has been compared with the Organon NucliSens assay on 521 retrospective samples obtained from HIV-1-positive patients monitored during highly active antiretroviral therapy, 79 of whom were assayed also by the Chiron Quantiplex 3.0 system and on characterized panels. The LCx system showed a moderate correlation (r = 0.795) and gave higher results than the NucliSens system on 245 of 327 concordant positive samples, with similar sensitivity. Correlation with Quantiplex system results was higher (r = 0.943). LCx reproducibility was very good; the procedure was simple, well controlled, and rapid (up to 48 results in 7 h). The HIV RNA quantitative assay on the LCx system is suitable for routine use. PMID- 11015430 TI - Histoplasmosis of the thyroid. AB - Fungal infection of the thyroid is rare. Most reported cases have involved Aspergillus, Coccidioides, and Candida species in the setting of disseminated disease. Infection of the thyroid with Histoplasma capsulatum is rarely reported as part of disseminated disease, even in geographic areas where histoplasmosis is endemic. We report a 52-year-old woman with a previous Hashimoto's disease and non-Hodgkin's lymphoma in which a diffuse enlarged thyroid gland with a large nodule was the only apparent locus of histoplasmosis. Fine-needle aspiration of the thyroid was an important diagnostic tool in establishing the diagnosis of histoplasmosis of the thyroid. The patient was initially treated with itraconazole (400 mg/day) for the fungal infection and six cycles of chemotherapy for the lymphoma. At a 6-month follow-up examination, the patient was doing well on suppressive therapy of itraconazole (200 mg/day), with no symptoms and with regression of the thyroid nodule and cervical adenopathy. PMID- 11015431 TI - Granulomatous amebic encephalitis in a patient with AIDS: isolation of acanthamoeba sp. Group II from brain tissue and successful treatment with sulfadiazine and fluconazole. AB - A patient with AIDS, treated with highly active antiretroviral therapy and trimethoprim-sulfamethoxazole, presented with confusion, a hemifield defect, and a mass lesion in the right occipital lobe. A brain biopsy confirmed granulomatous amebic encephalitis (GAE) due to Acanthamoeba castellanii. The patient was treated with fluconazole and sulfadiazine, and the lesion was surgically excised. This is the first case of AIDS-associated GAE responding favorably to therapy. The existence of a solitary brain lesion, absence of other sites of infection, and intense cellular response in spite of a very low CD4 count conditioned the favorable outcome. We review and discuss the diagnostic microbiologic options for the laboratory diagnosis of infections due to free-living amebae. PMID- 11015432 TI - Mycobacterium branderi from both a hand infection and a case of pulmonary disease. AB - Mycobacterium branderi, a potential human pathogen first characterized in 1995, has been isolated from respiratory tract specimens. We report here a case in which M. branderi was the only organism isolated upon culture from a hand infection. This isolate, along with a second isolate from a bronchial specimen, was subjected to conventional identification tests for mycobacterial species. Further analysis by high-performance liquid chromatography (HPLC) of mycolic acids and 16S rRNA gene sequencing was performed, and the antibiotic susceptibility profile was determined for both strains. Biochemical tests and the HPLC pattern were consistent with that of M. branderi and M. celatum, which are very similar. The 16S rRNA gene sequence of both strains corresponded to that of M. branderi and enabled us to confidently differentiate this organism from other closely related species such as M. celatum. This contributes to a further understanding of the status of this species as a potential human pathogen as well as illustrating the need for molecular diagnostics as a complementary method for the identification of rare mycobacterial species. PMID- 11015433 TI - Failure to detect circulating Aspergillus markers in a patient with chronic granulomatous disease and invasive aspergillosis. AB - We report a patient with chronic granulomatous disease who developed invasive pulmonary aspergillosis and a subphrenic abscess. During treatment, high levels of Aspergillus antigen were detected in the abscess, but circulating antigen and Aspergillus DNA were undetectable in the serum. PMID- 11015434 TI - Natural killer T cell activation inhibits hepatitis B virus replication in vivo. AB - We have previously reported that hepatitis B virus (HBV)-specific CD8(+) cytotoxic T lymphocytes and CD4(+) helper T lymphocytes can inhibit HBV replication in the liver of HBV transgenic mice by secreting interferon (IFN) gamma when they recognize viral antigen. To determine whether an activated innate immune system can also inhibit HBV replication, in this study we activated natural killer T (NKT) cells in the liver of HBV transgenic mice by a single injection of alpha-galactosylceramide (alpha-GalCer), a glycolipid antigen presented to Valpha14(+)NK1.1(+) T cells by the nonclassical major histocompatibility complex class I-like molecule CD1d. Within 24 h of alpha GalCer injection, IFN-gamma and IFN-alpha/beta were detected in the liver of HBV transgenic mice and HBV replication was abolished. Both of these events were temporally associated with the rapid disappearance of NKT cells from the liver, presumably reflecting activation-induced cell death, and by the recruitment of activated NK cells into the organ. In addition, prior antibody-mediated depletion of CD4(+) and CD8(+) T cells from the mice did not diminish the ability of alpha GalCer to trigger the disappearance of HBV from the liver, indicating that conventional T cells were not downstream mediators of this effect. Finally, the antiviral effect of alpha-GalCer was inhibited in mice that are genetically deficient for either IFN-gamma or the IFN-alpha/beta receptor, indicating that most of the antiviral activity of alpha-GalCer is mediated by these cytokines. Based on these results, we conclude that alpha-GalCer inhibits HBV replication by directly activating NKT cells and by secondarily activating NK cells to secrete antiviral cytokines in the liver. In view of these findings, we suggest that, if activated, the innate immune response, like the adaptive immune response, has the potential to control viral replication during natural HBV infection. In addition, the data suggest that therapeutic activation of NKT cells may represent a new strategy for the treatment of chronic HBV infection. PMID- 11015435 TI - Germinal center initiation, variable gene region hypermutation, and mutant B cell selection without detectable immune complexes on follicular dendritic cells. AB - Serum antibody (Ab) can play several roles during B cell immune responses. Among these is to promote the deposition of immune complexes (ICs) on follicular dendritic cells (FDCs). ICs on FDCs are generally thought to be critical for normal germinal center (GC) formation and the development and selection of memory B cells. However, it has been very difficult to test these ideas. To determine directly whether FDC-bound complexes do indeed function in these roles, we have developed a transgenic (Tg) mouse in which all B lymphocytes produce only the membrane-bound form of immunoglobulin M. Immune Tg mice have 10,000-fold less specific Ab than wild-type mice and lack detectable ICs on FDCs. Nonetheless, primary immune responses and the GC reaction in these mice are robust, suggesting that ICs on FDCs do not play critical roles in immune response initiation and GC formation. Moreover, as indicated by the presence and pattern of somatic mutations, memory cell formation and selection appear normal in these IC deficient GCs. PMID- 11015436 TI - Control of gammaherpesvirus latency by latent antigen-specific CD8(+) T cells. AB - The contribution of the latent antigen-specific CD8(+) T cell response to the control of gammaherpesvirus latency is currently obscure. Some latent antigens induce potent T cell responses, but little is known about their induction or the role they play during the establishment of latency. Here we used the murine gammaherpesvirus system to examine the expression of the latency-associated M2 gene during latency and the induction of the CD8(+) T cell response to this protein. M2, in contrast to the M3 latency-associated antigen, was expressed at day 14 after infection but was undetectable during long-term latency. The induction of the M2(91-99)/K(d) CD8(+) T cell response was B cell dependent, transient, and apparently induced by the rapid increase in latently infected cells around day 14 after intranasal infection. These kinetics were consistent with a role in controlling the initial "burst" of latently infected cells. In support of this hypothesis, adoptive transfer of an M2-specific CD8(+) T cell line reduced the initial load of latently infected cells, although not the long term load. These data represent the first description of a latent antigen specific immune response in this model, and suggest that vaccination with latent antigens such as M2 may be capable of modulating latent gammaherpesvirus infection. PMID- 11015437 TI - Attenuation of apoptosis underlies B lymphocyte stimulator enhancement of humoral immune response. AB - B lymphocyte stimulator (BLyS) is a newly identified monocyte-specific TNF family cytokine. It has been implicated in the development of autoimmunity, and functions as a potent costimulator with antiimmunoglobulin M in B cell proliferation in vitro. Here we demonstrate that BLyS prominently enhances the humoral responses to both T cell-independent and T cell-dependent antigens, primarily by attenuation of apoptosis as evidenced by the prolonged survival of antigen-activated B cells in vivo and in vitro. BLyS acts on primary splenic B cells autonomously, and directly cooperates with CD40 ligand (CD40L) in B cell activation in vitro by protecting replicating B cells from apoptosis. Moreover, although BLyS alone cannot activate the cell cycle, it is sufficient to prolong the survival of naive resting B cells in vitro. Attenuation of apoptosis by BLyS correlates with changes in the ratios between Bcl-2 family proteins in favor of cell survival, predominantly by reducing the proapoptotic Bak and increasing its prosurvival partners, Bcl-2 and Bcl-xL. In either resting or CD40L-activated B cells, the NF-kappaB transcription factors RelB and p50 are specifically activated, suggesting that they may mediate BLyS signals for B cell survival. Together, these results provide direct evidence for BLyS enhancement of both T cell-independent and T cell-dependent humoral immune responses, and imply a role for BLyS in the conservation of the B cell repertoire. The ability of BLyS to increase B cell survival indiscriminately, at either a resting or activated state, and to cooperate with CD40L, further suggests that attenuation of apoptosis underlies BLyS enhancement of polyclonal autoimmunity as well as the physiologic humoral immune response. PMID- 11015438 TI - CD1b-mediated T cell recognition of a glycolipid antigen generated from mycobacterial lipid and host carbohydrate during infection. AB - T cells recognize microbial glycolipids presented by CD1 proteins, but there is no information regarding the generation of natural glycolipid antigens within infected tissues. Therefore, we determined the molecular basis of CD1b-restricted T cell recognition of mycobacterial glycosylated mycolates, including those produced during tissue infection in vivo. Transfection of the T cell receptor (TCR) alpha and beta chains from a glucose monomycolate (GMM)-specific T cell line reconstituted GMM recognition in TCR-deficient T lymphoblastoma cells. This TCR-mediated response was highly specific for natural mycobacterial glucose-6-O (2R, 3R) monomycolate, including the precise structure of the glucose moiety, the stereochemistry of the mycolate lipid, and the linkage between the carbohydrate and the lipid. Mycobacterial production of antigenic GMM absolutely required a nonmycobacterial source of glucose that could be supplied by adding glucose to media at concentrations found in mammalian tissues or by infecting tissue in vivo. These results indicate that mycobacteria synthesized antigenic GMM by coupling mycobacterial mycolates to host-derived glucose. Specific T cell recognition of an epitope formed by interaction of host and pathogen biosynthetic pathways provides a mechanism for immune response to those pathogenic mycobacteria that have productively infected tissues, as distinguished from ubiquitous, but innocuous, environmental mycobacteria. PMID- 11015439 TI - Interferon gamma signaling alters the function of T helper type 1 cells. AB - One mechanism regulating the ability of different subsets of T helper (Th) cells to respond to cytokines is the differential expression of cytokine receptors. For example, Th2 cells express both chains of the interferon gamma receptor (IFN gammaR), whereas Th1 cells do not express the second chain of the IFN-gammaR (IFN gammaR2) and are therefore unresponsive to IFN-gamma. To determine whether the regulation of IFN-gammaR2 expression, and therefore IFN-gamma responsiveness, is important for the differentiation of naive CD4(+) T cells into Th1 cells or for Th1 effector function, we generated mice in which transgenic (TG) expression of IFN-gammaR2 is controlled by the CD2 promoter and enhancer. CD4(+) T cells from IFN-gammaR2 TG mice exhibit impaired Th1 polarization potential in vitro. TG mice also display several defects in Th1-dependent immunity in vivo, including attenuated delayed-type hypersensitivity responses and decreased antigen-specific IFN-gamma production. In addition, TG mice mount impaired Th1 responses against Leishmania major, as manifested by increased parasitemia and more severe lesions than their wild-type littermates. Together, these data suggest that the sustained expression of IFN-gammaR2 inhibits Th1 differentiation and function. Therefore, the acquisition of an IFN-gamma-unresponsive phenotype in Th1 cells plays a crucial role in the development and function of these cells. PMID- 11015440 TI - Tec family kinases modulate thresholds for thymocyte development and selection. AB - Tec family kinases are implicated in T cell receptor (TCR) signaling, and combined mutation of inducible T cell kinase (Itk) and resting lymphocyte kinase (Rlk)/Txk in mice dramatically impairs mature T cell function. Nonetheless, mutation of these kinases still permits T cell development. While itk(-)(/)- mice exhibit mild reductions in T cells with decreased CD4/CD8 cell ratios, rlk(-)(/) itk(-)(/)- mice have improved total T cell numbers yet maintain decreased CD4/CD8 ratios. Using TCR transgenics and an in vitro thymocyte deletion model, we demonstrate that mutation of Tec kinases causes graded defects in thymocyte selection, leading to a switch from negative to positive selection in rlk(-)(/) itk(-)(/)- animals. The reduction in both positive and negative selection and decreased CD4/CD8 ratios correlates with decreased biochemical parameters of TCR signaling, specifically defects in capacitive Ca(2+) influx and activation of the mitogen-activated kinases extracellular signal-regulated kinase 1 and 2. Thus, Tec kinases influence cell fate determination by modulating TCR signaling, leading to altered thresholds for thymocyte selection. These results provide support for a quantitative model for thymic development and provide evidence that defects in negative selection can substantially alter thymic cellularity. PMID- 11015441 TI - Reactive oxygen species (ROS)-induced ROS release: a new phenomenon accompanying induction of the mitochondrial permeability transition in cardiac myocytes. AB - We sought to understand the relationship between reactive oxygen species (ROS) and the mitochondrial permeability transition (MPT) in cardiac myocytes based on the observation of increased ROS production at sites of spontaneously deenergized mitochondria. We devised a new model enabling incremental ROS accumulation in individual mitochondria in isolated cardiac myocytes via photoactivation of tetramethylrhodamine derivatives, which also served to report the mitochondrial transmembrane potential, DeltaPsi. This ROS accumulation reproducibly triggered abrupt (and sometimes reversible) mitochondrial depolarization. This phenomenon was ascribed to MPT induction because (a) bongkrekic acid prevented it and (b) mitochondria became permeable for calcein ( approximately 620 daltons) concurrently with depolarization. These photodynamically produced "triggering" ROS caused the MPT induction, as the ROS scavenger Trolox prevented it. The time required for triggering ROS to induce the MPT was dependent on intrinsic cellular ROS-scavenging redox mechanisms, particularly glutathione. MPT induction caused by triggering ROS coincided with a burst of mitochondrial ROS generation, as measured by dichlorofluorescein fluorescence, which we have termed mitochondrial "ROS-induced ROS release" (RIRR). This MPT induction/RIRR phenomenon in cardiac myocytes often occurred synchronously and reversibly among long chains of adjacent mitochondria demonstrating apparent cooperativity. The observed link between MPT and RIRR could be a fundamental phenomenon in mitochondrial and cell biology. PMID- 11015442 TI - Carbon monoxide generated by heme oxygenase 1 suppresses endothelial cell apoptosis. AB - Heme oxygenase 1 (HO-1) inhibits apoptosis by regulating cellular prooxidant iron. We now show that there is an additional mechanism by which HO-1 inhibits apoptosis, namely by generating the gaseous molecule carbon monoxide (CO). Overexpression of HO-1, or induction of HO-1 expression by heme, protects endothelial cells (ECs) from apoptosis. When HO-1 enzymatic activity is blocked by tin protoporphyrin (SnPPIX) or the action of CO is inhibited by hemoglobin (Hb), HO-1 no longer prevents EC apoptosis while these reagents do not affect the antiapoptotic action of bcl-2. Exposure of ECs to exogenous CO, under inhibition of HO-1 activity by SnPPIX, substitutes HO-1 in preventing EC apoptosis. The mechanism of action of HO-1/CO is dependent on the activation of the p38 mitogen activated protein kinase (MAPK) signaling transduction pathway. Expression of HO 1 or exposure of ECs to exogenous CO enhanced p38 MAPK activation by TNF-alpha. Specific inhibition of p38 MAPK activation by the pyridinyl imidazol SB203580 or through overexpression of a p38 MAPK dominant negative mutant abrogated the antiapoptotic effect of HO-1. Taken together, these data demonstrate that the antiapoptotic effect of HO-1 in ECs is mediated by CO and more specifically via the activation of p38 MAPK by CO. PMID- 11015443 TI - Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. AB - PD-1 is an immunoinhibitory receptor expressed by activated T cells, B cells, and myeloid cells. Mice deficient in PD-1 exhibit a breakdown of peripheral tolerance and demonstrate multiple autoimmune features. We report here that the ligand of PD-1 (PD-L1) is a member of the B7 gene family. Engagement of PD-1 by PD-L1 leads to the inhibition of T cell receptor-mediated lymphocyte proliferation and cytokine secretion. In addition, PD-1 signaling can inhibit at least suboptimal levels of CD28-mediated costimulation. PD-L1 is expressed by antigen-presenting cells, including human peripheral blood monocytes stimulated with interferon gamma, and activated human and murine dendritic cells. In addition, PD-L1 is expressed in nonlymphoid tissues such as heart and lung. The relative levels of inhibitory PD-L1 and costimulatory B7-1/B7-2 signals on antigen-presenting cells may determine the extent of T cell activation and consequently the threshold between tolerance and autoimmunity. PD-L1 expression on nonlymphoid tissues and its potential interaction with PD-1 may subsequently determine the extent of immune responses at sites of inflammation. PMID- 11015444 TI - Salmonella-induced caspase-2 activation in macrophages: a novel mechanism in pathogen-mediated apoptosis. AB - The enterobacterial pathogen Salmonella induces phagocyte apoptosis in vitro and in vivo. These bacteria use a specialized type III secretion system to export a virulence factor, SipB, which directly activates the host's apoptotic machinery by targeting caspase-1. Caspase-1 is not involved in most apoptotic processes but plays a major role in cytokine maturation. We show that caspase-1-deficient macrophages undergo apoptosis within 4-6 h of infection with invasive bacteria. This process requires SipB, implying that this protein can initiate the apoptotic machinery by regulating components distinct from caspase-1. Invasive Salmonella typhimurium targets caspase-2 simultaneously with, but independently of, caspase 1. Besides caspase-2, the caspase-1-independent pathway involves the activation of caspase-3, -6, and -8 and the release of cytochrome c from mitochondria, none of which occurs during caspase-1-dependent apoptosis. By using caspase-2 knockout macrophages and chemical inhibition, we establish a role for caspase-2 in both caspase-1-dependent and -independent apoptosis. Particularly, activation of caspase-1 during fast Salmonella-induced apoptosis partially relies on caspase-2. The ability of Salmonella to induce caspase-1-independent macrophage apoptosis may play a role in situations in which activation of this protease is either prevented or uncoupled from the induction of apoptosis. PMID- 11015445 TI - Recruitment of SLP-76 to the membrane and glycolipid-enriched membrane microdomains replaces the requirement for linker for activation of T cells in T cell receptor signaling. AB - Two hematopoietic-specific adapters, src homology 2 domain-containing leukocyte phosphoprotein of 76 kD (SLP-76) and linker for activation of T cells (LAT), are critical for T cell development and T cell receptor (TCR) signaling. Several studies have suggested that SLP-76 and LAT function coordinately to promote downstream signaling. In support of this hypothesis, we find that a fraction of SLP-76 localizes to glycolipid-enriched membrane microdomains (GEMs) after TCR stimulation. This recruitment of SLP-76 requires amino acids 224-244. The functional consequences of targeting SLP-76 to GEMs for TCR signaling are demonstrated using a LAT/SLP-76 chimeric protein. Expression of this construct reconstitutes TCR-inducted phospholipase Cgamma1 phosphorylation, extracellular signal-regulated kinase activation, and nuclear factor of activated T cells (NFAT) promoter activity in LAT-deficient Jurkat T cells (J.CaM2). Mutation of the chimeric construct precluding its recruitment to GEMs diminishes but does not eliminate its ability to support TCR signaling. Expression of a chimera that lacks SLP-76 amino acids 224-244 restores NFAT promoter activity, suggesting that if localized, SLP-76 does not require an association with Gads to promote T cell activation. In contrast, mutation of the protein tyrosine kinase phosphorylation sites of SLP-76 in the context of the LAT/SLP-76 chimera abolishes reconstitution of TCR function. Collectively, these experiments show that optimal TCR signaling relies on the compartmentalization of SLP-76 and that one critical function of LAT is to bring SLP-76 and its associated proteins to the membrane. PMID- 11015446 TI - DAP10 and DAP12 form distinct, but functionally cooperative, receptor complexes in natural killer cells. AB - Many of the activating receptors on natural killer (NK) cells are multisubunit complexes composed of ligand-binding receptors that are noncovalently associated with membrane-bound signaling adaptor proteins, including CD3zeta, FcstraightepsilonRIgamma, DAP12, and DAP10. Because the DAP10 and DAP12 genes are closely linked, expressed in NK cells, and have remarkably similar transmembrane segments, it was of interest to determine the specificity of their interactions with ligand-binding receptors and to examine their signaling properties. Despite their similarities, DAP10, DAP12, FcstraightepsilonRIgamma, and CD3zeta form specific receptor complexes with their ligand-binding partners in NK cells and transfectants. The transmembrane regions of DAP10 and DAP12 are sufficient to confer specific association with their partners. Although cross-linking of either DAP10- or DAP12-associated receptors has been shown to be sufficient to trigger NK cell-mediated cytotoxicity against Fc receptor-bearing cells, substantial synergy was observed in the induction of cytokine production when both receptors were engaged. Activation of the Syk/ZAP70 tyrosine kinases by the immunoreceptor tyrosine-based activation motif-containing DAP12 adaptor and of the phosphatidylinositol 3-kinase pathway by the YxNM-containing DAP10 adaptor may play an important role in the stimulation of NK cells and T cells. PMID- 11015447 TI - LL-37, the neutrophil granule- and epithelial cell-derived cathelicidin, utilizes formyl peptide receptor-like 1 (FPRL1) as a receptor to chemoattract human peripheral blood neutrophils, monocytes, and T cells. AB - We have previously shown that antimicrobial peptides like defensins have the capacity to mobilize leukocytes in host defense. LL-37 is the cleaved antimicrobial 37-residue, COOH-terminal peptide of hCAP18 (human cationic antimicrobial protein with a molecular size of 18 kD), the only identified member in humans of a family of proteins called cathelicidins. LL-37/hCAP18 is produced by neutrophils and various epithelial cells. Here we report that LL-37 is chemotactic for, and can induce Ca(2+) mobilization in, human monocytes and formyl peptide receptor-like 1 (FPRL1)-transfected human embryonic kidney 293 cells. LL-37-induced Ca(2+) mobilization in monocytes can also be cross desensitized by an FPRL1-specific agonist. Furthermore, LL-37 is also chemotactic for human neutrophils and T lymphocytes that are known to express FPRL1. Our results suggest that, in addition to its microbicidal activity, LL-37 may contribute to innate and adaptive immunity by recruiting neutrophils, monocytes, and T cells to sites of microbial invasion by interacting with FPRL1. PMID- 11015448 TI - Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2. AB - Monocyte recruitment to the central nervous system (CNS) is a necessary step in the development of pathologic inflammatory lesions in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. Monocyte chemoattractant protein (MCP)-1, a potent agonist for directed monocyte migration, has been implicated in the pathogenesis of EAE. Here we report that deficiency in CC chemokine receptor (CCR)2, the receptor for MCP-1, confers resistance to EAE induced with a peptide derived from myelin oligodendrocyte glycoprotein peptide 35-55 (MOGp35-55). CCR2(-/)- mice immunized with MOGp35-55 failed to develop mononuclear cell inflammatory infiltrates in the CNS and failed to increase CNS levels of the chemokines RANTES (regulated on activation, normal T cell expressed and secreted), MCP-1, and interferon (IFN)-inducible protein 10 (IP-10) as well the chemokine receptors CCR1, CCR2, and CCR5. Additionally, T cells from CCR2(-/)- immunized mice showed decreased antigen-induced proliferation and production of IFN-gamma compared with wild-type immunized controls, suggesting that CCR2 enhances the T helper cell type 1 immune response in EAE. These data indicate that CCR2 plays a necessary and nonredundant role in the pathogenesis of EAE. PMID- 11015450 TI - Sporadic primary pulmonary hypertension is associated with germline mutations of the gene encoding BMPR-II, a receptor member of the TGF-beta family. AB - BACKGROUND: Primary pulmonary hypertension (PPH), resulting from occlusion of small pulmonary arteries, is a devastating condition. Mutations of the bone morphogenetic protein receptor type II gene (BMPR2), a component of the transforming growth factor beta (TGF-beta) family which plays a key role in cell growth, have recently been identified as causing familial PPH. We have searched for BMPR2 gene mutations in sporadic PPH patients to determine whether the same genetic defect underlies the more common form of the disorder. METHODS: We investigated 50 unrelated patients, with a clinical diagnosis of PPH and no identifiable family history of pulmonary hypertension, by direct sequencing of the entire coding region and intron/exon boundaries of the BMPR2 gene. DNA from available parent pairs (n=5) was used to assess the occurrence of spontaneous (de novo) mutations contributing to sporadic PPH. RESULTS: We found a total of 11 different heterozygous germline mutations of the BMPR2 gene in 13 of the 50 PPH patients studied, including missense (n=3), nonsense (n=3), and frameshift (n=5) mutations each predicted to alter the cell signalling response to specific ligands. Parental analysis showed three occurrences of paternal transmission and two of de novo mutation of the BMPR2 gene in sporadic PPH. CONCLUSION: The sporadic form of PPH is associated with germline mutations of the gene encoding the receptor protein BMPR-II in at least 26% of cases. A molecular classification of PPH, based upon the presence or absence of BMPR2 mutations, has important implications for patient management and screening of relatives. PMID- 11015449 TI - Channelopathies: ion channel defects linked to heritable clinical disorders. AB - Electrical signals are critical for the function of neurones, muscle cells, and cardiac myocytes. Proteins that regulate electrical signalling in these cells, including voltage gated ion channels, are logical sites where abnormality might lead to disease. Genetic and biophysical approaches are being used to show that several disorders result from mutations in voltage gated ion channels. Understanding gained from early studies on the pathogenesis of a group of muscle diseases that are similar in their episodic nature (periodic paralysis) showed that these disorders result from mutations in a gene encoding a voltage gated Na(+) channel. Their characterisation as channelopathies has served as a paradigm for other episodic disorders. For example, migraine headache and some forms of epilepsy have been shown to result from mutations in voltage gated Ca(2+) channel genes, while long QT syndrome is known to result from mutations in either K(+) or Na(+) channel genes. This article reviews progress made in the complementary fields of molecular genetics and cellular electrophysiology which has led to a better understanding of voltage gated ion channelopathies in humans and mice. PMID- 11015451 TI - ATR-X mutations cause impaired nuclear location and altered DNA binding properties of the XNP/ATR-X protein. AB - Mutations in the XNP/ATR-X gene, located in Xq13.3, are associated with several X linked mental retardation syndromes, the best known being alpha thalassaemia with mental retardation (ATR-X). The XNP/ATR-X protein belongs to the family of SWI/SNF DNA helicases and contains three C2-C2 type zinc fingers of unknown function. Previous studies have shown that 65% of mutations of XNP have been found within the zinc finger domain (encoded by exons 7, 8, and the beginning of exon 9) while 35% of the mutations have been found in the helicase domain extending over 3 kb at the C-terminus of the protein. Although different types of mutations have been identified, no specific genotype-phenotype correlation has been found, suggesting that gene alteration leads to a loss of function irrespective of mutation type. Our aims were to understand the function of the XNP/ATR-X protein better, with specific attention to the functional consequences of mutations to the zinc finger domain. We used monoclonal antibodies directed against the XNP/ATR-X protein and performed immunocytochemical and western blot analyses, which showed altered or absent XNP/ATR-X expression in cells of affected patients. In addition, we used in vitro experiments to show that the zinc finger domain can mediate double stranded DNA binding and found that the DNA binding capacity of mutant forms in ATR-X patients is severely reduced. These data provide insights into the understanding of the functional significance of XNP/ATR-X mutations. PMID- 11015452 TI - Divergent outcomes of intrachromosomal recombination on the human Y chromosome: male infertility and recurrent polymorphism. AB - The Y chromosome provides a unique opportunity to study mutational processes within the human genome, decoupled from the confounding effects of interchromosomal recombination. It has been suggested that the increased density of certain dispersed repeats on the Y could account for the high frequency of causative microdeletions relative to single nucleotide mutations in infertile males. Previously we localised breakpoints of an AZFa microdeletion close to two highly homologous complete human endogenous retroviral sequences (HERV), separated by 700 kb. Here we show, by sequencing across the breakpoint, that the microdeletion occurs in register within a highly homologous segment between the HERVs. Furthermore, we show that recurrent double crossovers have occurred between the HERVs, resulting in the loss of a 1.5 kb insertion from one HERV, an event underlying the first ever Y chromosomal polymorphism described, the 12f2 deletion. This event produces a substantially longer segment of absolute homology and as such may result in increased predisposition to further intrachromosomal recombination. Intrachromosomal crosstalk between these two HERV sequences can thus result in either homogenizing sequence conversion or a microdeletion causing male infertility. This represents a major subclass of AZFa deletions. PMID- 11015453 TI - Mutation analysis of the spastin gene (SPG4) in patients with hereditary spastic paraparesis. AB - BACKGROUND: Hereditary spastic paraparesis is a genetically heterogeneous condition. Recently, mutations in the spastin gene were reported in families linked to the common SPG4 locus on chromosome 2p21-22. OBJECTIVES: To study a population of patients with hereditary spastic paraparesis for mutations in the spastin gene (SPG4) on chromosome 2p21-22. METHODS: DNA from 32 patients (12 from families known to be linked to SPG4) was analysed for mutations in the spastin gene by single strand conformational polymorphism analysis and sequencing. All patients were also examined clinically. RESULTS: Thirteen SPG4 mutations were identified, 11 of which are novel. These mutations include missense, nonsense, frameshift, and splice site mutations, the majority of which affect the AAA cassette. We also describe a nucleotide substitution outside this conserved region which appears to behave as a recessive mutation. CONCLUSIONS: Recurrent mutations in the spastin gene are uncommon. This reduces the ease of mutation detection as a part of the diagnostic work up of patients with hereditary spastic paraparesis. Our findings have important implications for the presumed function of spastin and schemes for mutation detection in HSP patients. PMID- 11015454 TI - Dipeptidyl carboxypeptidase 1 (DCP1) and butyrylcholinesterase (BCHE) gene interactions with the apolipoprotein E epsilon4 allele as risk factors in Alzheimer's disease and in Parkinson's disease with coexisting Alzheimer pathology. AB - Alzheimer's disease (AD) and Parkinson's disease (PD) are genetically heterogeneous. Dipeptidyl carboxypeptidase 1 (DCP1) and butyrylcholinesterase (BCHE) genes may modify the risk of these disorders. We investigated whether common polymorphisms present in these genes operate as risk factors for AD and PD in Finnish subjects, independently or in concert with the apolipoprotein E epsilon4 allele (APOE epsilon4). Eighty late onset sporadic AD patients, 53 PD patients (34 of whom had concomitant AD pathology), and 67 control subjects were genotyped for the insertion (I)/deletion (D) polymorphism of DCP1 and the K variant of BCHE. In logistic regression analysis, the DCP1 *I allele in combination with APOE epsilon4 significantly increased the risk of AD (OR 30.0, 95% CI 7.3-123.7), compared to subjects carrying neither of the alleles. Similar analysis showed that the risk of AD was significantly increased in subjects carrying both the BCHE wild type (*WT/*WT) genotype and epsilon4 (OR 9.9, 95% CI 2.9-33.8), compared to those without this BCHE genotype and epsilon4. Further, the risk of PD with AD pathology was significantly increased for carriers of DCP1 *I and epsilon4 (OR 8.0, 95% CI 2.1-31.1). We thus conclude that, in Finns, interaction between DCP1 *I and epsilon4 increases the risk of AD as well as of PD with coexisting Alzheimer pathology, which underlines the importance of the DCP1 I/D polymorphism in the development of Alzheimer neuropathology, whereas the wild type BCHE genotype in combination with epsilon4 had a combined effect with regard to the risk of AD. PMID- 11015455 TI - Two translocations of chromosome 15q associated with dyslexia. AB - Developmental dyslexia is characterised by difficulties in learning to read. As reading is a complex cognitive process, multiple genes are expected to contribute to the pathogenesis of dyslexia. The genetics of dyslexia has been a target of molecular studies during recent years, but so far no genes have been identified. However, a locus for dyslexia on chromosome 15q21 (DYX1) has been established in previous linkage studies. We have identified two families with balanced translocations involving the 15q21-q22 region. In one family, the translocation segregates with specific dyslexia in three family members. In the other family, the translocation is associated with dyslexia in one family member. We have performed fluorescence in situ hybridisation (FISH) studies to refine the position of the putative dyslexia locus further. Our results indicate that both translocation breakpoints on 15q map within an interval of approximately 6-8 Mb between markers D15S143 and D15S1029, further supporting the presence of a locus for specific dyslexia on 15q21. PMID- 11015456 TI - Two common forms of the human MLH1 gene may be associated with functional differences. PMID- 11015457 TI - FMR3 is a novel gene associated with FRAXE CpG island and transcriptionally silent in FRAXE full mutations. AB - We have identified a novel gene, FMR3, originating from the FRAXE CpG island. The FMR3 gene is transcribed from the opposite strand to the FMR2 gene. Analogous to the silencing of the FMR1 and FMR2 genes, FMR3 transcription is extinguished by FRAXE full mutation. Although the role of FMR3 in FRAXE associated mild to borderline mental retardation is not yet clear, lack of expression of FMR3 in FRAXE full mutation males means that the FMR3 gene is potentially involved. PMID- 11015458 TI - Genetics and deafness: what do families want? PMID- 11015459 TI - A case of dyschondrosteosis from Roman Britain. PMID- 11015460 TI - Xp;Yp translocation inherited from the father in an SRY, RBM, and TSPY positive true hermaphrodite with oligozoospermia. PMID- 11015461 TI - MPS II in females: molecular basis of two different cases. PMID- 11015462 TI - Partial tetrasomy 21 in a male infant. PMID- 11015463 TI - Benign familial infantile convulsions: report of a UK family and confirmation of genetic heterogeneity. PMID- 11015464 TI - Low frequency of microsatellite instability in BRCA1 mutated breast tumours. PMID- 11015465 TI - Effects of tea consumption on nutrition and health. AB - Beneficial health effects of tea have been demonstrated in animal experiments and some human studies. The two most extensively investigated diseases are cancer and heart disease. Although mechanisms of protective activity of tea against these diseases have been proposed, there are inconsistencies in the relationship between tea consumption and the risk of these diseases in humans. The bioavailability of active components is beginning to be understood, but further research is required to determine whether the results from animal studies are applicable to humans. Also discussed are the possible effects of tea in increasing thermogenesis and bone density as well as decreasing risk of cataracts and arthritis. The potential health benefits of tea consumption warrant further investigation. PMID- 11015466 TI - Leucine stimulates translation initiation in skeletal muscle of postabsorptive rats via a rapamycin-sensitive pathway. AB - The objectives of the present study were twofold: 1) to determine whether leucine is unique among the branched-chain amino acids (BCAA) in its ability to stimulate protein synthesis in skeletal muscle of food-deprived rats; and 2) to investigate whether changes in muscle protein synthesis after leucine administration involve a signaling pathway that includes the protein kinase mammalian target of rapamycin (mTOR). In the first set of experiments, food-deprived (18 h) male rats (200 g) were orally administered saline or 270 mg valine, isoleucine or leucine. In the second set of experiments, food-deprived rats were injected intravenously with rapamycin (0.75 mg/kg), a specific inhibitor of mTOR, before leucine administration. Only leucine stimulated protein synthesis in skeletal muscle above saline-treated controls (P: < 0.05). Furthermore, leucine was most effective among the BCAA at enhancing phosphorylation of eukaryotic initiation factor (eIF), 4E binding protein 1 (4E-BP1) and the 70-kDa ribosomal protein S6 kinase (S6K1). Leucine-dependent hyperphosphorylation of 4E-BP1 increased the availability of eIF4E to form the active eIF4G.eIF4E complex. To a lesser extent, isoleucine also enhanced phosphorylation of 4E-BP1 and S6K1. Rapamycin inhibited protein synthesis in both leucine-treated and food-deprived rats. Additionally, rapamycin prevented the stimulatory effects of leucine on eIF4E availability for binding eIF4G and inhibited leucine-dependent phosphorylation of S6K1. The data demonstrate that leucine is unique among the BCAA in its ability to stimulate protein synthesis in muscle of food-deprived rats. We show for the first time that leucine-dependent stimulation of translation initiation in vivo occurs via a rapamycin-sensitive pathway. PMID- 11015467 TI - Palm tocotrienols protect ApoE +/- mice from diet-induced atheroma formation. AB - We evaluated the effects of vitamin E and beta-carotene on apolipoprotein (apo)E +/- female mice, which develop atherosclerosis only when fed diets high in triglyceride and cholesterol. Mice were fed a nonpurified control diet (5.3 g/100 g triglyceride, 0.2 g/100 g cholesterol), an atherogenic diet alone (15.8 g/100 g triglyceride, 1.25 g/100 g cholesterol, 0.5 g/100 g Na cholate) or the atherogenic diet supplemented with either 0.5 g/100 g (+)-alpha-tocopherol (mixed isomers); 0.5 g/100 g palm tocopherols (palm-E; 33% alpha-tocopherol, 16.1% alpha tocotrienol, 2.3% beta-tocotrienol, 32.2% gamma-tocotrienol, 16.1% delta tocotrienol); 1.5 g/100 g palm-E; or 0.01 g/100 g palm-carotenoids (58% beta carotene, 33% alpha-carotene, 9% other carotenoids). Compared with mice fed the control diet, plasma cholesterol was fourfold greater in mice fed the atherogenic diet. Mice fed the 1.5 g/100 g palm-E supplement had 60% lower plasma cholesterol than groups fed the other atherogenic diets. Mice fed the atherogenic diet had markedly higher VLDL, intermediate density lipoprotein (IDL) and LDL cholesterol and markedly lower HDL cholesterol than the controls. Lipoprotein patterns in mice supplemented with alpha-tocopherol or palm carotenoids were similar to those of the mice fed the atherogenic diet alone, but the pattern in mice supplemented with 1. 5 g/100 g palm-E was similar to that of mice fed the control diet. In mice fed the atherogenic diet, the hepatic cholesterol plus cholesterol ester concentration was 4.4-fold greater than in mice fed the control diet. Supplementing with 1.5 g/100 g palm-E lowered hepatic cholesterol plus cholesterol ester concentration 66% compared with the atherogenic diet alone. Mice fed the atherogenic diet had large atherosclerotic lesions at the level of the aortic valve. With supplements of 0.5 g/100 g palm-E or 1.5 g/100 g palm-E, the size of the lesions was 92 or 98% smaller, respectively. The 0.5 g/100 g alpha-tocopherol and palm carotenoid supplements had no effect. Supplements did not alter mRNA abundance for apolipoproteins A1, E, and C3. The beneficial effect of tocotrienols on atherogenesis, the plasma lipoprotein profile and accumulation of hepatic cholesterol esters cannot be attributed to their antioxidant properties. PMID- 11015468 TI - Dietary fat modulates serum paraoxonase 1 activity in rats. AB - We examined the effects of dietary fats with specific fatty acid compositions, on serum paraoxonase (PON1) activity in rats. Male adult Sprague-Dawley rats were divided randomly into four dietary groups. One group received the control diet [AIN 93M with soybean oil (5 g/100 g diet)], whereas the remaining three groups received the modified control diet supplemented with (15 g/100 g diet) triolein, tripalmitin or fish oil, respectively. After 20 d, blood was obtained after overnight food deprivation and PON1 activity was determined. Serum lipids and lipid components of lipoproteins were also determined. Serum PON1 activity [micromol/(L.min)] was significantly (P: < 0.05) higher in triolein (98 +/- 6) and lower in fish oil (41 +/- 4), compared with tripalmitin-fed rats (63 +/- 11). Serum PON1 activity in tripalmitin-fed rats was comparable to that of controls (67 +/- 9). Serum PON1 activity correlated significantly with serum lecithin:cholesterol acyltransferase (LCAT) activity (r = 0.77, P: < 0.001) and was transported in blood principally in association with the denser subfraction of HDL, very high density lipoprotein (VHDL; d > 1.15 kg/L). Serum PON1 activity correlated strongly with serum lipids as well as lipids of VLDL, HDL and its subfractions. Multiple linear regression analysis, however, showed a significant relationship of serum PON1 activity, principally with the phospholipids of VHDL (r = 0.47, P: < 0.002). These data suggest that the modulation of serum PON1 activity by dietary fat may be mediated via the effect of the specific fatty acids on the synthesis and secretion of VHDL, the subfraction of HDL that transports the majority of PON1 in the blood. PMID- 11015469 TI - Highly unsaturated (n-3) fatty acids, but not alpha-linolenic, conjugated linoleic or gamma-linolenic acids, reduce tumorigenesis in Apc(Min/+) mice. AB - We showed previously that dietary eicosapentaenoic acid [EPA, 20:5(n-3)] is antitumorigenic in the APC:(Min/+) mouse, a genetic model of intestinal tumorigenesis. Only a few studies have evaluated the effects of dietary fatty acids, including EPA and docosahexaenoic acid [DHA, 22:6(n-3)], in this animal model and none have evaluated the previously touted antitumorigenicity of alpha linolenic acid [ALA, 18:3(n-3)], conjugated linoleic acid [CLA, 77% 18:2(n-7)], or gamma-linolenic acid [GLA, 18:3(n-6)]. Stearidonic acid [SDA, 18:4(n-3)], the Delta6-desaturase product of ALA, which is readily metabolized to EPA, has not been evaluated previously for antitumorigenic efficacy. This study was undertaken to evaluate the antitumorigenicity of these dietary fatty acids (ALA, SDA, EPA, DHA, CLA and GLA) compared with oleic acid [OA, 18:1(n-9)] at a level of 3 g/100 g in the diets of APC:(Min/+) mice and to determine whether any alterations in tumorigenesis correspond to alterations in prostaglandin biosynthesis. Tumor multiplicity was significantly lower by approximately 50% in mice fed SDA or EPA compared with controls, whereas less pronounced effects were observed in mice fed DHA (P: = 0.15). ALA, CLA and GLA were ineffective at the dose tested. Although lower tumor numbers coincided with significantly lower prostaglandin levels in SDA- and EPA-fed mice, ALA and DHA supplementation resulted in equally low prostaglandin levels, despite proving less efficacious with regard to tumor number. Prostaglandin levels did not differ significantly in the CLA and GLA groups compared with controls. These results suggest that SDA and EPA attenuate tumorigenesis in this model and that this effect may be related in part to alterations in prostaglandin biosynthesis. PMID- 11015470 TI - Acute starvation and subsequent refeeding affect lymphocyte subsets and proliferation in cats. AB - Although the early identification of patients with suboptimal nutritional status can allow the implementation of nutritional intervention to enhance the ability of the body to fight infection and disease, currently no definitive test of nutritional status exists. Therefore, this study was conducted to identify possible functional indicators of acute nutritional deprivation. The effects of total nutritional deprivation and subsequent refeeding on lymphocyte functions and subpopulations were examined in 23 healthy cats. Peripheral blood samples were analyzed at various times during food deprivation and refeeding periods. During the food deprivation period, decreases were observed in leukocyte number (P: < 0.05), lymphocyte number (P: < 0.05), percentage of CD4(+) cells [before stimulation with concanavalin-A (Con-A); P: < 0.05] and the CD4/CD8 ratio (before stimulation with Con-A; P: < 0.01) compared with d 0. Increases were observed in the percentage of CD8(+) cells [before (P: < 0.05) and after (P: < 0.01) stimulation with Con-A] and in intracellular calcium (P: < 0.01) during acute starvation. During the refeeding period, increases were observed in the percentage of CD4(+) cells (before and after stimulation with Con-A; P: < 0.01), the percentage of CD8(+) cells (before stimulation with Con-A; P: < 0.05) and lymphocyte number (P: < 0.05) compared with d 7. Lymphocyte proliferative capacity tended to decrease (P: = 0.07) during starvation and increased (P: < 0.01) during the refeeding period. These findings suggest that a 7-d starvation period had immunosuppressive effects on cats and that these effects were not completely normalized during 7 d of refeeding. CD4(+)/CD8(+) subset alterations and CD4/CD8 ratio in conjunction with lymphocyte proliferation may be useful as indices of nutritional status. PMID- 11015471 TI - Fats infused intraduodenally affect the postprandial secretion of the exocrine pancreas and the plasma concentration of cholecystokinin but not of peptide YY in growing pigs. AB - In pigs, the spontaneous secretion of the exocrine pancreas and the release of cholecystokinin (CCK) and peptide YY (PYY) after intraduodenal infusion of fully saturated synthetic fats differing in chain length was studied. Growing pigs (n = 6) were prepared with pancreatic duct catheters, duodenal T-cannulas and catheters placed in the jugular vein. The pigs were fed 2 g/100 g body twice daily. Beginning with the morning feeding, a medium-chain triglyceride (MCT: glycerol tricaprylate), a long-chain triglyceride (LCT: glycerol tristearate) or saline was infused at a rate of 0.1 g/100 g body. Pancreatic juice was collected, beginning 1 h preprandially until 3 h postprandially. Blood samples were obtained 15 min preprandially and 15, 45, 90 and 150 min postprandially. The infusion of MCT evoked a change in the trend of the curve for the volume of secretion of pancreatic juice, lipase and colipase concentrations and outputs. The trend of the curve did not change over time for CCK and PYY. Differences between the trends of the curves for the saline and MCT treatment were observed for volume of secretion, protein output, lipase content and output, trypsin and colipase output. Differences in the trends of the curves between MCT and LCT were obtained for the outputs of protein, lipase and colipase. Plasma CCK levels were lower as a result of the MCT treatment compared with the saline and LCT treatments. The results suggest an immediate, distinguished response of the porcine exocrine pancreas to fats differing in chain length. PMID- 11015472 TI - Dietary fructans modulate polyamine concentration in the cecum of rats. AB - Nondigestible but fermentable dietary fructans such as oligofructose exert many effects on gut physiology through their fermentation end products such as short chain fatty acids. Could other metabolites be produced in the gut and contribute to the physiologic effects of dietary fructans? The aim of the study was to evaluate the influence of oligofructose on putrescine, spermidine and spermine concentrations in the cecum, the portal vein and the liver of rats and to assess their involvement in cecal enlargement and the modulation of hepatic lipid metabolism. Putrescine, spermidine and spermine were quantified by HPLC in samples obtained from male Wistar rats fed a nonpurified standard diet (controls) or the same diet enriched with 10 g/100 g oligofructose (OFS) for 4 wk. OFS-fed rats had significantly greater cecal content and tissue weights. OFS almost doubled the concentration of putrescine in the cecal contents. The concentration of all three polyamines in the cecal tissue was significantly greater than in controls. The concentration of spermidine in portal plasma was lower in rats fed OFS, whereas the treatment did not affect the polyamine concentrations in the liver. The fermentation of dietary fructans contributed to an increase in the concentration of putrescine in the gut without modifying putrescine concentration in either the portal blood or liver. Moreover, the greater levels of polyamines in cecal tissue may be related to the cell proliferation resulting from OFS fermentation in the gut. PMID- 11015473 TI - Food deprivation increases alpha(2)-adrenoceptor-mediated modulation of jejunal epithelial transport in young and adult rats. AB - This study examined the effect of food deprivation on the jejunal response to alpha(2)-adrenoceptor activation in young (20-d-old) and adult (60-d-old) rats, using short-circuit (I(sc)) measurements in the absence or presence of furosemide (1 mmol/L). The effect of alpha(2)-adrenoceptor stimulation by 5-bromo-N:-(4, 5 dihydro-1H-imidazol-2-yl)-6-quinoxalinamine (UK 14,304; 0.3-3000 nmol/L) was a concentration-dependent decrease in I(sc) with similar half-maximal effective concentration (EC(50); 12.3 +/- 1.1 vs. 9.6 +/- 1.1 nmol/L) and maximal effect (E(max); 70.6 +/- 6.9 vs. 80.6 +/- 4.5% of reduction) values in adult food deprived and fed rats. The effect of UK 14,304 on I(sc) in fed and food-deprived rats was markedly (P: < 0.05) attenuated by furosemide (1 mmol/L). E(max) values for UK 14,304 in 20-d-old food-deprived rats were higher (P: < 0.05) than those observed in fed rats (93.3 +/- 3.3 vs. 67.0 +/- 11.3% of reduction), without differences in EC(50) values. The effect of UK 14,304 on I(sc) in 20-d-old fed rats was completely abolished by furosemide (1 mmol/L). In food-deprived young rats, the effect of UK 14,304 was also markedly (P: < 0.05) antagonized by furosemide, but not completely abolished. Specific [(3)H]-rauwolscine binding in membranes from jejunal epithelial cells revealed the presence of a single class of binding sites, with an apparent K:(D) in the low nmol/L range. In 20-d-old food-deprived rats, specific [(3)H]-rauwolscine binding was markedly increased, and this was reversed by refeeding. Na(+),K(+)-ATPase activity in isolated jejunal epithelial cells from 60-d-old fed rats was twice that in 20-d-old fed rats [117 +/- 14 vs. 52 +/- 5 nmol free inorganic phosphorus/(mg protein.min)]. Food deprivation in adult rats, but not in 20-d-old rats, was accompanied by a significant decrease in Na(+),K(+)-ATPase activity. In both young and adult rats (fed and food-deprived), UK 14,304 did not affect Na(+),K(+)-ATPase activity. In conclusion, food deprivation in 20-d-old rats enhanced the response to alpha(2) adrenoceptor stimulation. This effect, which depends primarily on the stimulation of a furosemide-sensitive antisecretory mechanism, is suggested to result from increases in the number of jejunal epithelial alpha(2)-adrenoceptors. PMID- 11015474 TI - Kinetics of carnitine palmitoyltransferase-I are altered by dietary variables and suggest a metabolic need for supplemental carnitine in young pigs. AB - To examine the kinetics of carnitine palmitoyltransferase-I (CPT-I) and the influence of dietary variables, young pigs (18 kg, n = 20) were fed corn-soybean meal diets supplemented with 40 g soy oil/kg and containing either 136 or 180 g crude protein/kg and either 0 or 500 mg/kg L-carnitine (2 x 2 factorial design). Diets were offered for 10 d (85% of ad libitum); CPT-I activities in liver and skeletal muscle mitochondria were determined, and enzyme kinetic constants (V:(max) and K:(m) for carnitine) were estimated. Kinetics of CPT-I in muscle were not affected by diet (P: > 0.1; carnitine K:(m) = 480 +/- 44 micromol/L). In contrast, the K:(m) for carnitine in liver was increased from 164 to 216 +/- 20 micromol/L by dietary L-carnitine supplementation (P: < 0.01) and from 169 to 211 +/- 20 micromol/L by high protein feeding (P: < 0.05). Dietary L-carnitine increased muscle and liver free carnitine concentrations by 72 and 158% over control concentrations (770 and 80 micro;mol/kg wet muscle and liver, respectively). Because tissue carnitine concentrations were within the range of the respective K:(m) for both liver and muscle tissue, it is inferred that alteration of tissue carnitine concentrations via dietary supplementation could modulate CPT-I activity in young pigs. PMID- 11015475 TI - Conjugated linoleic acid persistently increases total energy expenditure in AKR/J mice without increasing uncoupling protein gene expression. AB - AKR/J mice fed a high fat diet were treated with a 1% (1 g/100 g) admixture of conjugated linoleic acids (CLA) for 5 wk and compared with control mice. Body weights, energy intakes and energy expenditure (EE) determined by indirect calorimetry were measured weekly. CLA treatment reduced adipose depot weights by approximately 50% but had no significant effects on either body weight or energy intake. CLA increased EE persistently by an average of 7.7% throughout the 5-wk experiment. This greater EE, despite no difference in energy intake, was sufficient to account for the lower body fat stores in the CLA-treated mice. De novo fatty acid biosynthesis in adipose tissue, measured by incorporation of deuterium-labeled water, was not decreased by CLA treatment and therefore did not explain the lower adipose lipid in these mice. Expression of uncoupling protein (UCP) in skeletal muscle, white adipose tissue and kidney was not affected by CLA treatment. In brown adipose tissue, UCP1 expression was not affected by CLA treatment. However, UCP2 expression, although quite low, was significantly greater in CLA-fed mice. We conclude that CLA acts to reduce body fat stores by chronically increasing metabolic rate. This effect on metabolic rate is likely not due to increased UCP gene expression. Furthermore, the reduced body fat is not due to decreased de novo fatty acid synthesis in white adipose tissue. PMID- 11015476 TI - Relationship between human adipose tissue agouti and fatty acid synthase (FAS). AB - The human homologue of the murine obesity gene, agouti, is expressed in adipose tissue. We have shown that recombinant agouti protein regulates adipocyte lipogenesis and lipolysis coordinately and promotes lipid storage via a Ca(2+) dependent mechanism in vitro, which may contribute to agouti-induced obesity. However, little is known about agouti's physiologic function in humans. We first studied the agouti content in human mature adipocytes vs. preadipocytes. The agouti content of human mature adipocytes was five times as abundant as in preadipocytes (19.18 +/- 2.46 vs. 4.07 +/- 0.51 pg/microg protein, P: < 0.005), suggesting that agouti is up-regulated during adipocyte differentiation. We next studied the relationship of agouti mRNA and protein to fatty acid synthase (FAS) mRNA and activity in adipose tissue obtained from nonobese and mildly obese patients (body mass index range, 21-31 kg/m(2)). Agouti protein was correlated with FAS activity (r = 0.782, P: < 0.005). Similarly, human adipose tissue agouti mRNA level was also correlated with FAS mRNA level (r = 0.846, P: < 0.001). These data suggest that agouti may be another adipocyte-produced factor that modulates adipocyte lipid metabolism via a paracrine/autocrine mechanism. PMID- 11015477 TI - Growth hormone receptor gene expression in porcine skeletal and cardiac muscles is selectively regulated by postnatal undernutrition. AB - During mild postnatal undernutrition, growth hormone receptor (GHR) mRNA abundance decreases in liver but increases in longissimus dorsi muscle. We tested the following hypotheses: 1) GHR gene expression is related to the metabolic and contractile characteristics of different muscles, and 2) the GHR response to nutrition depends on muscle type. Eight pairs of littermate pigs were weaned at 3 wk and given an optimal [60 g/(kg.d)] or low [(20 g/(kg.d)] food intake for the next 3 wk. All pigs grew, but at a slower rate in the low food intake group (P: < 0.001). Functionally distinct muscles were assessed for GHR mRNA (RNase protection analysis), oxidative myofibers (succinate dehydrogenase histochemistry) and type I slow myofibers (myosin immunocytochemistry). There were striking muscle-specific differences in GHR gene expression (P: < 0.001) and in its regulation by nutritional status. Relative expression of GHR mRNA in the optimal food intake group occurred in ascending order as follows: longissimus < diaphragm approximately rhomboideus < cardiac < soleus. There was a positive correlation with the proportion of oxidative myofibers (P: < 0.001) but not with type I myofibers (P: > 0.10). Compared with the high intake pigs, hepatic GHR mRNA was downregulated in the low intake pigs by 59% (P: < 0.01), whereas in the four muscles examined it was upregulated as follows: longissimus, 124% (P: < 0.05); rhomboideus, 19% (P: > 0.4); soleus, 65% (P: < 0. 05); cardiac, 51% (P: < 0.05). Moreover, the proportion of skeletal muscle fibers with high oxidative capacity was also greater in the low intake group (P: < 0.05). We conclude that postnatal GHR gene expression and its regulation by mild undernutrition are related to the metabolic, contractile and specific functional properties of different muscles. PMID- 11015478 TI - Genistein activates apolipoprotein A-I gene expression in the human hepatoma cell line Hep G2. AB - Soy phytoestrogens have been shown to increase plasma levels of HDL cholesterol and apolipoprotein (apo) A-I, its major protein component, in animal studies and in some human studies. The human hepatoma cell line Hep G2 was used to study the effect of the phytoestrogens genistein and daidzein on apo A-I secretion and gene expression in liver cells. Both genistein and daidzein increased apo A-I secretion in a dose-dependent fashion. Apo A-I concentration in the media of treated cells was increased approximately fivefold by 10 micromol/L genistein (P: < 0.001) and approximately onefold by 10 micromol/L daidzein (P: < 0.001) compared with control cells. The effect of genistein on apo A-I secretion was similar to that observed with 17-beta-estradiol. Treatment of cells with genistein for 24 h increased the transcriptional activity of the apo A-I gene as measured by nuclear run-on assay. Transfection experiments with plasmids containing regulatory regions of the apo A-I gene cloned in front of the luciferase reporter gene indicated that the 5' region of the apo A-I gene contained between nucleotides -256 and -41 is responsible for the increased expression of this gene by genistein. PMID- 11015479 TI - The association between two common mutations C677T and A1298C in human methylenetetrahydrofolate reductase gene and the risk for diabetic nephropathy in type II diabetic patients. AB - Mutations of the methylenetetrahydrofolate reductase (MTHFR) gene have been shown to be associated with a predisposition to developing diabetic nephropathy (DN) in specific populations. The frequency of two MTHFR mutations, a recently described mutation in the human MTHFR gene A1298C and C677T, whose association with DN is already known, was determined in an Israeli Jewish population with type 2 diabetes mellitus (DM). Both A1298C and C677T are highly prevalent in the diabetic population with allele frequencies of 0.35 and 0.36, respectively. The genotype frequency and allele frequency for these two polymorphisms in patients who are normoalbuminuric (n = 55) were compared with those of patients who had either micro- or macroalbuminuria (n = 43). For both polymorphisms, there were no significant differences in either the genotype distribution or allele frequency in patients with or without DN. However, in patients with serum folate <15.4 nmol/L, there was a greater incidence of DN in those patients who were homozygous or heterozygous for the C677T mutation. For the A1298C mutation, there is evidence suggesting that the homozygous state may be protective in patients with low-normal serum folate. Folate supplementation in diabetic patients with the C677T mutation and low-normal serum folate may prevent the onset or retard the progression of DN. PMID- 11015480 TI - Growth velocity, fat-free mass and energy intake are inversely related to viral load in HIV-infected children. AB - The study objectives were to assess the relationships among human immunodeficiency virus (HIV) replication, energy balance, body composition and growth in children with HIV-associated growth failure (GF). Energy intake and expenditure, body composition and level of HIV RNA were measured in 16 HIV infected children with growth failure (HIV+/GF+), defined as a 12-mo height velocity 20% and the quadratic malnutrition gap by >30%, but would be very expensive to implement. "Broad stroke" geographic targeting could reduce the same measures by 15 and 20%, respectively, and would be much less expensive to implement. We conclude that geographic targeting has the potential to substantially enhance the effect of nutrition programs on the severity of stunting in Honduras. PMID- 11015484 TI - Dietary vitamin A intake and nondietary factors are associated with reversal of stunting in children. AB - We examined prospectively the associations between dietary vitamin A intake, nondietary factors and growth in 8174 Sudanese children ages 6-72 mo who were stunted at the start of follow-up. All subjects were weighed and measured at baseline and at 6-mo intervals for 18 mo of follow-up. Dietary vitamin A intake during the prior 24 h was assessed using recall of vitamin A-containing foods at baseline and 6-mo intervals. We examined the association of dietary vitamin A intake with growth and the incidence of recovery of stunting after controlling for age, sex, breast-feeding status and socioeconomic variables. We found that carotenoid intake was associated with a greater incidence of reversal of stunting. Children in the highest quintile grew 13 mm more during the study period than children in the lowest quintile [95% confidence interval (CI): 0-25 mm] in multivariate analyses. The relative risk (RR) of recovery associated with vitamin A intake was greater in infants up to 1 y old (RR = 3.3, CI: 0.9-11.7) than in children > or =3 y of age (RR = 1.0, CI: 0.8-1. 3) (P:-value for interaction = 0.08). Diets rich in carotenoids may increase the rate of recovery from stunting in children. Dietary effects on growth might be strongest among very young children and those who have been most malnourished. Age, sex, breast feeding status, socioeconomic status and severity of baseline stunting also were associated with reversal of stunting in this population. PMID- 11015485 TI - Hookworms, malaria and vitamin A deficiency contribute to anemia and iron deficiency among pregnant women in the plains of Nepal. AB - Anemia and iron deficiency during pregnancy are prevalent in developing countries, but their causes are not always known. We assessed the prevalence and severity of anemia and iron deficiency and their association with helminths, malaria and vitamin A deficiency in a community-based sample of 336 pregnant women in the plains of Nepal. Hemoglobin, erythrocyte protoporphyrin (EP) and serum ferritin were assessed in venous blood samples. Overall, 72.6% of women were anemic (hemoglobin < 110 g/L), 19.9% had moderate to severe anemia (hemoglobin < 90 g/L) and 80.6% had iron deficiency (EP > 70 micromol/mol heme or serum ferritin < 10 microg/L). Eighty-eight percent of cases of anemia were associated with iron deficiency. More than half of the women (54.2%) had a low serum retinol concentration (<1.05 micromol/L), 74.2% were infected with hookworms and 19.8% had Plasmodium vivax malaria parasitemia. Hemoglobin, EP and serum ferritin concentrations were significantly worse and the prevalence of anemia, elevated EP and low serum ferritin was increased with increasing intensity of hookworm infection. Hookworm infection intensity was the strongest predictor of iron status, especially of depleted iron stores. Low serum retinol was most strongly associated with mild anemia, whereas P. vivax malaria and hookworm infection intensity were stronger predictors of moderate to severe anemia. These findings reinforce the need for programs to consider reducing the prevalence of hookworm, malaria infection and vitamin A deficiency where indicated, in addition to providing iron supplements to effectively control anemia. PMID- 11015487 TI - Soy protein peptides regulate cholesterol homeostasis in Hep G2 cells. AB - The activation of LDL receptors was described recently in a human hepatoma cell line (Hep G2) exposed both to alpha + alpha' subunits from 7S soy globulin and to Croksoy(R)70, a commercial isoflavone-poor soy concentrate. To assess the final identity of the peptide(s) putatively responsible for the biochemical effect, experiments were performed in Hep G2 cells, exposed either to synthetic peptides corresponding to specific sequences of 7S soy globulin or to peptides from the in vitro digestion of Croksoy(R)70. Moreover, the ability of the whole 7S globulin, its subunits and whole Croksoy(R)70 to interfere in the apolipoprotein B (apo B) secretion in the medium as well as in sterol biosynthesis was evaluated in the same model. Increased (125)I-LDL uptake and degradation vs. controls were shown after Hep G2 incubation with a synthetic peptide (10(-)(4) mol/L, MW 2271 Da) corresponding to positions 127-150 of the 7S globulin. Cells exposed to Croksoy(R)70 enzyme digestion products showed a more marked up-regulation of LDL receptors vs. controls, compared with vs. Hep G2 cells incubated with undigested Croksoy(R)70. Among soy-derived products, only the 7S globulin inhibited apo B secretion and (14)C-acetate incorporation when tested in Hep G2 cells at a concentration of 1.0 g/L. These findings support the hypothesis that if one or more peptides can reach the liver after intestinal digestion, they may elicit a cholesterol-lowering effect. Moreover, the protein moiety, devoid of isoflavone components, is likely to be responsible for this major biochemical effect of soy protein. PMID- 11015486 TI - Tomato intake in relation to mortality and morbidity among Sudanese children. AB - The intake of foods that contain high levels of antioxidants may counteract the adverse effects of oxidative stress and lead to improved immune function and reduced risk of infectious disease. We prospectively examined the relationship between the consumption of tomatoes, a rich source of antioxidants, and mortality and diarrheal and respiratory morbidity rates among 28,753 children who were 6-60 mo old and enrolled in a longitudinal study in the Sudan. Children in each household were visited every 6 mo for a maximum of four visits. At each round, mothers recalled whether a child had consumed tomatoes in the previous 24 h. Events (death or morbidity) reported at each round were prospectively allocated according to the number of days of tomato intake. Intake of tomatoes for 2 or 3 d compared with none was associated, respectively, with 48% (relative risk, 0. 53; 95% confidence interval, 0.30-0.91) and 83% (0.17; 0.04-0.72) reductions in morality rates (P: for trend = 0.002). The association between tomato use and death remained statistically significant (P: for trend = 0.004), even after further adjustment for total vitamin A intake. Tomato intake was also associated with a reduced risk of death associated with diarrhea in the week preceding death (P: for trend = 0.009) or fever (P: for trend = 0.04). Intake of tomatoes was also inversely and significantly associated with the risks of diarrheal and respiratory infections. Our data suggest that tomatoes may be beneficial for child health but also emphasize the general importance of food-based approaches to the prevention of micronutrient malnutrition and protection of the health of children in developing countries. PMID- 11015488 TI - Incorporation and metabolism of dietary trans isomers of linolenic acid alter the fatty acid profile of rat tissues. AB - To study the influence on lipid metabolism and platelet aggregation of the fatty acid isomerization that occurs during heat treatment, weanling rats were fed for 8 wk a diet enriched with 5% isomerized (experimental group) or normal (control group) canola oil. Geometrical isomers of alpha-linolenic acid representing 0.2 g/100 g of the experimental diet were incorporated into liver, platelets, aorta and heart, at the expense of their cis homologue and of 18:2(n-6). The major isomer, 9c,12c,15t-18:3, was also metabolized to 5c,8c,11c,14c,17t-20:5 and to an unknown compound, found in liver, platelets and aorta, which has been identified tentatively as 7c, 10c,13c,16c,19t-22:5. The greater 20:4(n-6)/18:2(n-6) ratio in the liver, platelets and heart of the experimental group than the control group indicated an enhancement of desaturation activities. This induced a higher content of long-chain (n-6) fatty acids in the experimental group. Platelet aggregation tended to be slightly higher (P: = 0.065) in the experimental group. We conclude that 0.2 g of trans isomers of alpha-linolenic acid per 100 g of diet was sufficient to be incorporated and metabolized, thus altering the fatty acid profile of rat tissues. PMID- 11015489 TI - Riboflavin phosphorylation is the crucial event in riboflavin transport by isolated rat enterocytes. AB - In isolated rat enterocytes, both normoenergized (normal) and de-energized with rotenone, riboflavin intracellular metabolic processes, operating in association with a membrane-specific transport mechanism, were investigated. The contents of unlabeled (endogenous) and labeled (exogenous) flavins [riboflavin (RF), flavin mononucleotide (FMN), flavin adenindinucleotide (FAD)] were determined by HPLC before and after incubation with tritiated RF. In normoenergized enterocytes, total labeled RF content (i.e., total uptake, the sum of RF membrane transport and intracellular metabolism) increased steadily to a plateau after 20 min incubation; FMN and FAD contents reached a plateau between 3 and 20 min, whereas free RF content increased constantly. The phosphorylated forms prevailed over the free form ( approximately 60% of total flavins). In de-energized enterocytes, RF total uptake was significantly lower than in normoenergized enterocytes and reached a plateau after only 3 min incubation. FMN and FAD contents were significantly lower than in normoenergized enterocytes, and free RF represented the prevailing form of flavins (70% of total RF ). In both normoenergized and de energized enterocytes, the contents of unlabeled total RF, FMN and FAD decreased significantly after 20 min incubation, whereas free RF increased significantly only in normoenergized enterocytes. After 20 min incubation, the RF structural analog 8-dimethyl-amino-8-demethyl-RF caused a significant decrease of all flavin contents, whereas 5'-deoxy-RF decreased only the total and free RF contents. Results directly confirmed the leading role of metabolic processes such as phosphorylation in RF transport by isolated small intestinal enterocytes. PMID- 11015490 TI - The diurnal rhythm of energy expenditure differs between obese and glucose intolerant rats and streptozotocin-induced diabetic rats. AB - Otsuka Long Evans Tokushima Fatty (OLETF) rats were developed as a model of noninsulin-dependent diabetes mellitus (NIDDM) with mild obesity. Changes in carcass composition and in the daily profile of energy expenditure were examined before and after manifestation of diabetes (8 and 24 wk, respectively), and compared with the normal control Long Evans Tokushima (LETO) rats and streptozotocin (STZ)-induced diabetic LETO rats. OLETF rats had greater body weights than LETO rats and significantly greater absolute and relative fat weights. A diurnal rhythm of energy expenditure associated with two peaks was observed in LETO rats, but the two peaks were not apparent in OLETF rats at 24 wk of age. A diurnal rhythm associated with one peak was observed in STZ-induced diabetic LETO rats. Energy derived from fat constituted this peak; the pattern of the daily energy expenditure was significantly different from that of either nontreated LETO or OLETF rats at 24 wk of age. NIDDM in OLETF rats at 24 wk of age has only a small role in modification of the diurnal rhythm of energy expenditure, whereas STZ-induced diabetes significantly affected the rhythm. PMID- 11015491 TI - Mammary lipogenic enzyme activity, trans fatty acids and conjugated linoleic acids are altered in lactating dairy cows fed a milk fat-depressing diet. AB - The objectives of the present study were to examine the effect of a milk fat depressing (MFD) diet on: 1) the activity of mammary acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS), 2) ACC mRNA relative abundance and 3) distributions of conjugated linoleic acids (CLA) and trans-18:1 fatty acids (tFA) in milk fat. Twelve lactating Holstein cows were used in a single reversal design. Two diets were fed: a control diet (60:40% forage/concentrate) and an MFD diet (25:70% forage/concentrate, supplemented with 5% soybean oil). The MFD diet decreased (P: < 0 0.001) milk fat by 43% and ACC and FAS activity by 61 and 44%, respectively. A reduced ACC mRNA relative abundance (P: < 0.001) corresponded with the lower ACC activity. The fatty acids synthesized de novo were decreased (P: < 0. 002), whereas tFA were increased from 1.9 to 15.6% due predominantly to a change in trans-10-18:1 isomer (P: < 0.001). With the MFD diet, the trans-7, cis-9 and trans-10, cis-12 CLA isomers were elevated (P: < 0.001), in contrast to the decrease in trans-11-18:1 (P: < 0. 001) and cis-9, trans-11-18:2. The data were consistent with a dietary effect on mammary de novo FA synthesis mediated through a reduction in ACC and FAS activity and in ACC mRNA abundance. The results were compatible with a role of trans-10, cis-12 CLA in milk fat depression, but alterations noted in tFA and other CLA isomers suggest that they also may be important during diet-induced milk fat depression. PMID- 11015492 TI - Boron supplementation of a semipurified diet for weanling pigs improves feed efficiency and bone strength characteristics and alters plasma lipid metabolites. AB - Two experiments were conducted to determine effects of dietary boron (B) on performance, plasma minerals and metabolites, and bone characteristics in young pigs. In Experiment 1, 48 pigs (24 males, 24 females; 21 d old) were allotted to pens, which were randomly assigned to one of the following dietary treatments: 1) control (natural ingredient diet; 6.7 mg B/kg diet), 2) control + 5 mg B/kg diet and 3) control + 15 mg B/kg diet. Boron was supplemented as sodium borate. In Experiment 2, 48 pigs (24 males, 24 females; 21 d old) were assigned to the same treatments described in Experiment 1; however, the basal diet was a semipurified diet (0.98 mg B/kg diet). Diets were fed for 40 d; on d 40, blood samples were obtained for determination of plasma mineral and metabolite concentrations. Femurs were harvested from 8 pigs per treatment on d 40 for determination of mechanical properties, ash and lipid percentage. In Experiment 1, B did not affect performance, plasma minerals or metabolites or bone properties. In Experiment 2, B supplementation improved (P: < 0.05) the gain:feed ratio and increased plasma cholesterol and triglyceride concentrations. There was a treatment x sex interaction (P: < 0.05) in Experiment 2 for bone lipid to be lower and bending moment to be higher, with the response occurring in male pigs. Other dependent variables in Experiment 2 were not affected by treatment. In conclusion, B supplementation of a low B diet elicited responses of physiologic importance to pigs. However, B supplementation of a natural ingredient diet did not elicit a response. PMID- 11015493 TI - Hypothalamic nuclei are malformed in weanling offspring of low protein malnourished rat dams. AB - Maternal low protein malnutrition during gestation and lactation (LP) is an animal model frequently used for the investigation of long-term deleterious consequences of perinatal growth retardation. Both perinatal malnutrition and growth retardation at birth are risk factors for diabetic and cardiovascular disturbances in later life. The pathophysiologic mechanisms responsible are unknown. Hypothalamic nuclei are decisively involved in the central nervous regulation of food intake, body weight and metabolism. We investigated effects of a low protein diet (8% protein; control diet, 17% protein) during gestation and lactation in rat dams on the organization of hypothalamic regulators of body weight and metabolism in the offspring at weaning (d 20 of life). LP offspring had significantly lower body weight than control offspring (CO; P: < 0.001), associated with hypoglycemia and hypoinsulinemia (P: < 0. 005) on d 20 of life. This was accompanied by a greater relative volume of the ventromedial hypothalamic nucleus (P: < 0.01) and a greater numerical density of Nissl-stained neurons in this nucleus (P: < 0.01) as well as in the paraventricular hypothalamic nucleus (PVN; P: < 0.001). In contrast, no significant differences in neuronal densities were observed generally in the lateral hypothalamic area, arcuate hypothalamic nucleus (ARC), and dorsomedial hypothalamic nucleus between LP offspring and CO offspring. On the other hand, LP offspring displayed fewer neurons immunopositive for neuropeptide Y in the ARC (P: < 0.05), whereas in the PVN, lower neuronal densities of neurons immunopositive for galanin were found in LP offspring compared with CO offspring (P: < 0.001). On the contrary, in the PVN, no significant group difference in the numerical density of cholecystokinin 8S-positive neurons was present. A long-term effect of these specific hypothalamic alterations on body weight and metabolism in LP offspring during later life is suggested. PMID- 11015494 TI - Intake of soy products is associated with better plasma lipid profiles in the Hong Kong Chinese population. AB - We describe the pattern of soy intake and its association with blood lipid concentrations in the Hong Kong Chinese population. Subjects were contacted by random telephone survey and invited to a hospital for a physical examination and blood tests. A total of 500 men and 510 women with an age range of 24-74 y completed the dietary intake study. The dietary assessment was based on a semiquantitative food frequency questionnaire that included 10 commonly consumed soy items. Many (88%) of the study population had consumed some soy products during the previous week. About 80% of the soy protein or isoflavones were obtained from different forms of tofu, and an additional 9% was obtained from soy milk. The mean weekly isoflavone intake was 102 +/- 107 mg in men and 77 +/- 90 mg in women. In men, soy intake and total plasma cholesterol were negatively correlated (r = -0.09, P: = 0.04), as were soy intake and LDL cholesterol (r = 0.11, P: = 0.02). The respective values in women <50 y old were r = -0.11, P: = 0.04 and r = -0.11, P: = 0.05. Soy protein remained significantly associated with these two lipid concentrations after adjustment for other social and dietary confounders. Higher soy intake seemed to be related to a better plasma lipid profile in men and in younger women, but more epidemiological studies and controlled clinical trials in this setting would help to confirm the optimal amount required for the prevention and treatment of hyperlipidemia. PMID- 11015495 TI - Early weaning and prolonged nursing induce changes in cell proliferation in the gastric epithelium of developing rats. AB - Food deprivation stimulates cell proliferation in the gastric epithelium of suckling, but not weanling rats. This study was designed to investigate the role of diet on proliferation in developing animals, using early weaning and prolonged nursing models. Rat pups were subjected to these dietary conditions at d 15 and were killed 3 or 7 d afterwards. One day before killing, half of pups were deprived of food. Body weights were recorded. After mitosis blockade, the histologic sections of the stomach were used for the evaluation of cell proliferation and methapasic cell distribution along the gland, and for the measurement of mucosa thickness. Body weight was impaired at 18 d by early weaning and at 22 d by prolonged nursing. Food restriction promoted a 10-15% weight loss regardless of dietary conditions. At 18 d, food deprivation inhibited cell division (P: < 0.01) and reduced the thickness of the mucosa (P: < 0.05) in rats that were weaned early. At 22 d, only the thickness of the mucosa was different between the groups that were subjected to early weaning and prolonged nursing (P: < 0.05), regardless of feeding state. The frequency of dividing cells along the gland was affected by early weaning in 18- and 22-d-old rats. These results suggest the following: 1) food deprivation effects are dependent on dietary condition at 18 d because different proliferative responses were achieved after early weaning and prolonged nursing; 2) the lack of changes after dietary manipulation in 22-d-old rats indicates a nonresponsive period during postnatal development. We conclude that milk is a modulatory factor for cell proliferation in the gastric mucosa of rats. PMID- 11015496 TI - Fermentation by gut microbiota cultured in a simulator of the human intestinal microbial ecosystem is improved by supplementing a soygerm powder. AB - An in vitro model, designated the Simulator of the Human Intestinal Microbial Ecosystem (SHIME), was used to study the effect of a soygerm powder rich in beta glycosidic phytoestrogenic isoflavones on the fermentation pattern of the colon microbiota and to determine to what extent the latter metabolize the conjugated phytoestrogens. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 3 wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota were monitored. The microbiota were then subjected to a 2-wk treatment period by adding 2.5 g/d soygerm powder to the culture medium. The addition resulted into an overall increase of bacterial marker populations (Enterobacteriaceae:, coliforms, Lactobacillus: sp., Staphylococcus: sp. and Clostridium: sp.), with a significant increase of the Lactobacillus: sp. population. The short-chain fatty acid (SCFA) concentration increased approximately 30% during the supplementation period; this was due mainly to a significant increase of acetic and propionic acids. Gas analysis revealed that the methane concentration increased significantly. Ammonium and sulfide concentrations were not influenced by soygerm supplementation. Use of an electronic nose apparatus indicated that odor concentrations decreased significantly during the treatment period. The beta-glycosidic bonds of the phytoestrogenic isoflavones were cleaved under the conditions prevailing in the large intestine. The increased bacterial fermentation after addition of the soygerm powder was paralleled by substantial metabolism of the free isoflavones (genistein, daidzein and glycitein), resulting in recovery of only 12-17% of the supplemented isoflavones. PMID- 11015497 TI - Body weight-specific zinc compartmental masses in girls significantly exceed those reported in adults: a stable isotope study using a kinetic model. AB - Maintaining optimal zinc status is important for normal growth and development in children, but minimal data are available regarding zinc metabolism in this age group. Our objectives were to utilize stable isotope-based compartmental modeling techniques to investigate zinc metabolism in healthy children; to expand a current stable isotope-based model to include red blood cell data; and to compare kinetic parameters in children with those previously reported in adults. Seven healthy girls, age 9.94 +/- 0.79 y, received 1.1 mg of a (67)zinc-enriched tracer orally and 0.5 mg of a (70)zinc-enriched tracer intravenously. Blood, urine and fecal samples were collected for 6 d. Stable isotope enrichments were measured by thermal ionization magnetic sector mass spectrometry. A six-compartment model based on a model previously reported in adults was used; the model excluded red blood cell data. Body weight-corrected masses of the body zinc compartments derived using this model were significantly greater in children than those reported in adults. Modification of the model to include a red blood cell compartment increased the total identifiable zinc mass of the nongastrointestinal compartments by approximately 2.5%. We conclude that compartmental modeling can be used to describe zinc kinetics in children, and that the body weight-corrected zinc pool masses are significantly greater in children than in adults. PMID- 11015498 TI - Foodborne illnesses and nutritional status: a statement from an American Society for Nutritional Sciences Working Group. PMID- 11015499 TI - Use of bootstrap procedure and Monte Carlo simulation. PMID- 11015500 TI - Response to use of bootstrap procedure and monte carlo simulation PMID- 11015501 TI - Cerebral intravascular oxygenation correlates with mean arterial pressure in critically ill premature infants. AB - OBJECTIVES: Premature infants experience brain injury, ie, germinal matrix intraventricular hemorrhage (GMH-IVH) and periventricular leukomalacia (PVL), in considerable part because of disturbances in cerebral blood flow (CBF). Because such infants are susceptible to major fluctuations in mean arterial blood pressure (MAP), impaired cerebrovascular autoregulation would increase the likelihood for the changes in CBF that could result in GMH-IVH and PVL. The objectives of this study were to determine whether a state of impaired cerebrovascular autoregulation could be identified reliably and conveniently at the bedside, the frequency of any such impairment, and the relation of the impairment to the subsequent occurrence of severe GMH-IVH and PVL. PATIENTS AND METHODS: To monitor the cerebral circulation continuously and noninvasively, we used near-infrared spectroscopy (NIRS) to determine quantitative changes in cerebral concentrations of oxygenated hemoglobin (HbO(2)) and deoxygenated hemoglobin (Hb) from the first hours of life. Our previous experimental study showed a strong correlation between a measure of cerebral intravascular oxygenation (HbD), ie, HbD = HbO(2) - Hb, determined by NIRS, and volemic CBF, determined by radioactive microspheres. We studied 32 very low birth weight premature infants (gestational age: 23-31 weeks; birth weight: 605-1870 g) requiring mechanical ventilation, supplemental oxygen, and invasive blood pressure monitoring by NIRS from 1 to 3 days of age. MAP measured by arterial catheter pressure transducer and arterial oxygen saturation measured by pulse oximetry were recorded simultaneously. The relationship of MAP to HbD was quantitated by coherence analysis. RESULTS: Concordant changes (coherence scores >. 5) in HbD and MAP, consistent with impaired cerebrovascular autoregulation, were observed in 17 of the 32 infants (53%). Eight of the 17 infants (47%) developed severe GMH-IVH or PVL or both. Of the 15 infants with apparently intact autoregulation, ie, coherence scores <.5, only 2 (13%) developed severe ultrasonographic lesions. Thus, for the entire study population of 32 infants, 8 of the 10 with severe lesions exhibited coherence scores >.5. CONCLUSIONS: We conclude that NIRS can be used in a noninvasive manner at the bedside to identify premature infants with impaired cerebrovascular autoregulation, that this impairment is relatively common in such infants, and that the presence of this impairment is associated with a high likelihood of occurrence of severe GMH IVH/PVL. PMID- 11015502 TI - Adverse sedation events in pediatrics: analysis of medications used for sedation. AB - OBJECTIVES: To perform a systematic investigation of medications associated with adverse sedation events in pediatric patients using critical incident analysis of case reports. METHODS: One hundred eighteen case reports from the adverse drug reporting system of the Food and Drug Administration, the US Pharmacopoeia, and the results of a survey of pediatric specialists were used. Outcome measures were death, permanent neurologic injury, prolonged hospitalization without injury, and no harm. The overall results of the critical incident analysis are reported elsewhere. The current investigation specifically examined the relationship between outcome and medications: individual and classes of drugs, routes of administration, drug combinations and interactions, medication errors and overdoses, patterns of drug use, practitioners, and venues of sedation. RESULTS: Ninety-five incidents fulfilled study criteria and all 4 reviewers agreed on causation; 60 resulted in death or permanent neurologic injury. Review of adverse sedation events indicated that there was no relationship between outcome and drug class (opioids; benzodiazepines; barbiturates; sedatives; antihistamines; and local, intravenous, or inhalation anesthetics) or route of administration (oral, rectal, nasal, intramuscular, intravenous, local infiltration, and inhalation). Negative outcomes (death and permanent neurologic injury) were often associated with drug overdose (n = 28). Some drug overdoses were attributable to prescription/transcription errors, although none of 39 overdoses in 34 patients seemed to be a decimal point error. Negative outcomes were also associated with drug combinations and interactions. The use of 3 or more sedating medications compared with 1 or 2 medications was strongly associated with adverse outcomes (18/20 vs 7/70). Nitrous oxide in combination with any other class of sedating medication was frequently associated with adverse outcomes (9/10). Dental specialists had the greatest frequency of negative outcomes associated with the use of 3 or more sedating medications. Adverse events occurred despite drugs being administered within acceptable dosing limits. Negative outcomes were also associated with drugs administered by nonmedically trained personnel and drugs administered at home. Some injuries occurred on the way to a facility after administration of sedatives at home; some took place in automobiles or at home after discharge from medical supervision. Deaths and injuries after discharge from medical supervision were associated with the use of medications with long half-lives (chloral hydrate, pentobarbital, promazine, promethazine, and chlorpromazine). CONCLUSIONS: Adverse sedation events were frequently associated with drug overdoses and drug interactions, particularly when 3 or more drugs were used. Adverse outcome was associated with all routes of drug administration and all classes of medication, even those (such as chloral hydrate) thought to have minimal effect on respiration. Patients receiving medications with long plasma half-lives may benefit from a prolonged period of postsedation observation. Adverse events occurred when sedative medications were administered outside the safety net of medical supervision. Uniform monitoring and training standards should be instituted regardless of the subspecialty or venue of practice. Standards of care, scope of practice, resource management, and reimbursement for sedation should be based on the depth of sedation achieved (ie, the degree of vigilance and resuscitation skills required) rather than on the drug class, route of drug administration, practitioner, or venue. PMID- 11015504 TI - Neonatal outcome of preimplantation genetic diagnosis by polar body removal: the first 109 infants. AB - CONTEXT: Our center developed the technique of preimplantation genetic diagnosis (PGD) by sequential polar body removal (PBR) for the diagnosis of Mendelian disorders and aneuploidies. This study examines the obstetric and neonatal outcome of the first 109 live births after PGD by PBR. OBJECTIVE: To determine if there were any observable effects of PGD by PBR on perinatal morbidity and mortality, birth defects, and growth parameters. DESIGN: Data on perinatal outcome were gathered for the first 109 infants by parental reporting and confirmed by telephone interview and chart review when indicated. In infants >6 months old, a follow-up telephone interview was performed establishing the developmental milestones attained by the child. SETTING: A research center conducting an institutional review board-approved research protocol in PGD. PATIENTS: All patients who had PGD by PBR who had clinical pregnancies. MAIN OUTCOME MEASURES: Gestational age, mode of delivery, perinatal mortality, birth weight, birth length, the presence of birth defects, and developmental milestones. RESULTS: There was no significant decrease in birth length or weight, or the frequency of small for gestational age infants. No specific pattern of birth defects was observed. CONCLUSION: Thus far, there are no observable detrimental effects of PGD by PBR on children born after the procedure. PMID- 11015503 TI - The prevalence of genital human papillomavirus infections in abused and nonabused preadolescent girls. AB - OBJECTIVE: To compare the prevalence of genital human papillomavirus (HPV) infections in sexually abused and nonabused preadolescent girls and assess the feasibility of conducting a longitudinal study of the natural history of HPV infection in this population. METHOD: Consecutively referred, 5- to 12-year-old girls who were evaluated for sexual abuse by a Child Advocacy and Protection Team were invited to participate in the study. During a standard forensic medical examination, 2 specimens for HPV testing were obtained (one by rubbing a Dacron swab over the perineum and the other by lavaging the vagina with phosphate buffered saline). The specimens were evaluated for HPV DNA by polymerase chain reaction using MY09/11 consensus primers and high-risk (16,18,31,33,35,39,45,51,52, 56,58) and low-risk (6,11,42,43,44) types were detected with a solution hybridization assay, the SHARP Signal System (Digene Diagnostics). The genital area was examined for warts and subclinical, colposcopic evidence of HPV. Participants were invited to return for longitudinal evaluation at 4-month intervals for 2 years. RESULTS: Sexual abuse was confirmed in 29 (72.5%) of the 40 study participants, suspected in 2 (5%), and ruled out in 9 (22.5%). None of the girls had genital warts or abnormal colposcopic findings. HPV DNA was detected in 5 (16%) of the 31 girls with confirmed or suspected sexual abuse (1 with high-risk and 4 with low-risk types) and none of the nonabused girls (Fisher's exact test). Girls who tested positive and negative for HPV did not differ significantly in age or type of abuse. Despite close telephone follow-up and numerous attempts to schedule appointments, none of the participants returned for follow-up. CONCLUSIONS: Genital HPV infection is more common among sexually abused than nonsexually abused girls, with the majority of infections not clinically apparent. Because it is so difficult to study the natural history of these infections in abused children, it may be necessary to draw inferences about the long-term sequelae of pediatric HPV infections from longitudinal studies of girls who voluntarily initiate sexual activity soon after menarche. PMID- 11015505 TI - Video recording as a means of evaluating neonatal resuscitation performance. AB - OBJECTIVE: To determine the compliance to Neonatal Resuscitation Program (NRP) guidelines in our institution, by the use of videotaped newborn resuscitations. BACKGROUND: NRP is the standard of care for newborn resuscitation. The application of NRP guidelines and resuscitation skills in actual clinical settings is undocumented. DESIGN/METHODS: A video recorder, mounted to the radiant warmer in the main obstetrical operating room, was used to record all high-risk resuscitations. All members of the resuscitation team were NRP certified. The videotapes were reviewed within 14 days of the resuscitation and then erased. This ongoing review was approved as a quality assurance (QA) project ensuring confidentiality under California law. The first 100 resuscitations were evaluated to assess NRP compliance. Each step in the resuscitation (positioning, oxygen delivery, ventilation, chest compressions, intubation, and medication) was graded. A score was devised, with 2 points being awarded for every correct decision and proper procedure, 1 point for delayed interventions or inadequate technique, and zero points for indicated procedures that were omitted or for interventions that were not indicated. The total points were divided by the total possible points for that patient. The scores for the first 25 resuscitations (group 1) and the last 25 resuscitations (group 2) were compared. RESULTS: Fifty four percent of the 100 resuscitations had deviations from the NRP guidelines. Ten percent received overly aggressive stimulation and 22% had poor suction technique. Of the 78 infants given oxygen, this decision was considered incorrect in 15% and the delivery technique was poor in 10% of the infants given oxygen. Of those requiring mask ventilation (n = 18), 24% had poor chest expansion, 11% used an incorrect rate, and 17% had inadequate reevaluation. Twelve infants were intubated; only 7 were successfully intubated on the first attempt and only 4 were intubated in <20 seconds. The longest intubation attempt was 50 seconds. Naloxone was given to 2 patients. One was breathing spontaneously with a heart rate >100. Resuscitations receiving a perfect evaluation score were more likely to occur in infants needing less intervention. The level of resuscitation required for groups 1 and 2 were statistically similar. There was no difference in resuscitation scores between the 2 groups. Only the inappropriate use of deep suctioning improved, with 8 of 25 events in group 1, and 0 of 25 in group 2. CONCLUSIONS: We have found a significant number of deviations from the NRP guidelines. Video recording of actual clinical practice is a useful QA tool for monitoring the conduct of newborn resuscitation. We are now conducting repeat video assessments of individual NRP providers to determine whether there is improved performance. PMID- 11015506 TI - The EPICure study: outcomes to discharge from hospital for infants born at the threshold of viability. AB - OBJECTIVE: To evaluate the outcome for all infants born before 26 weeks of gestation in the United Kingdom and the Republic of Ireland. This report is of survival and complications up until discharge from hospital. METHODOLOGY: A prospective observational study of all births between March 1, 1995 and December 31, 1995 from 20 to 25 weeks of gestation. RESULTS: A total of 4004 births were recorded, and 811 infants were admitted for intensive care. Overall survival was 39% (n = 314). Male sex, no reported chorioamnionitis, no antenatal steroids, persistent bradycardia at 5 minutes, hypothermia, and high Clinical Risk Index for Babies (CRIB) score were all independently associated with death. Of the survivors, 17% had parenchymal cysts and/or hydrocephalus, 14% received treatment for retinopathy of prematurity (ROP), and 51% needed supplementary oxygen at the expected date of delivery. Failure to administer antenatal steroids and postnatal transfer for intensive care within 24 hours of birth were predictive of major scan abnormality; lower gestation was predictive of severe ROP, while being born to a black mother was protective. Being of lower gestation, male sex, tocolysis, low maternal age, neonatal hypothermia, a high CRIB score, and surfactant therapy were all predictive of oxygen dependency. Intensive care was provided in 137 units, only 8 of which had >5 survivors. There was no difference in survival between institutions when divided into quintiles based on their numbers of extremely preterm births or admissions. CONCLUSIONS: This study provides outcome data for this geographically defined cohort; survival and neonatal morbidity are consistent with previous data from the United Kingdom and facilitate comparison with other geographically based data. PMID- 11015507 TI - Costs and effectiveness of ultrasonography and limited computed tomography for diagnosing appendicitis in children. AB - BACKGROUND: A protocol of ultrasonography (US) followed by computed tomography with rectal contrast (CTRC) has been shown to be 94% accurate in the diagnosis of acute appendicitis in children. OBJECTIVE: To evaluate the changes in patient management and costs of a protocol using US and CTRC in the evaluation of appendicitis in children. DESIGN, SETTING, AND SUBJECTS: Prospective cohort study of 139 children between 3 and 21 years of age who had equivocal clinical findings for acute appendicitis seen in the emergency department of a large, urban pediatric teaching hospital between July 1998 and December 1998. PROTOCOL: Children with equivocal clinical presentations for acute appendicitis were prospectively evaluated with US. Patients with positive findings for acute appendicitis went directly to the operating room. Patients with negative or equivocal findings on US underwent CTRC. Surgical management plans were recorded before imaging, after US, and after CTRC. MAIN OUTCOME MEASURES: Surgical management plans before and after the imaging protocol as well as total hospital direct and indirect costs incurred or saved by each change in management were determined. Costs were obtained through the hospital's cost database and by ratios of costs to charges. RESULTS: Of the 139 children, the protocol resulted in a beneficial change in management in 86 children (61.9%), no change in management in 50 children (36.0%) and an incorrect change in management in 3 children (2.1%). US alone resulted in a beneficial change in management decision in 12/31 children (38.7%), while US followed by CTRC resulted in a beneficial change in management in 74/108 children (68.5%). The protocol resulted in a total cost savings of $78 503.99 or $565/patient. CONCLUSION: A protocol of US followed by CTRC in children with negative or equivocal US examinations results in a high rate of beneficial change in management as well as in total cost savings in children with equivocal clinical presentations for suspected appendicitis. PMID- 11015508 TI - Decline in prevalence of neural tube defects in a high-risk region of the United States. AB - OBJECTIVES: To conduct surveillance for neural tube defects (NTDs) in a high-risk region of the United States and to prevent occurrence and recurrence of NTDs through the periconceptional use of folic acid supplements. DESIGN: Active and passive methods were used for surveillance of NTD-affected pregnancies and births during a 6-year period (October 1992-September 1998). Individual genetic counseling was used to prevent NTD recurrences and a public awareness campaign was used to reduce NTD occurrences. SETTING: State of South Carolina. PATIENTS: All cases of spina bifida, anencephaly, and encephalocele identified among 278 122 live births and fetal deaths to South Carolina residents during 1992-1998 were included. MAIN OUTCOME MEASURE: Changes in occurrence and recurrence rates during a 6-year period. RESULTS: Over the 6 years of surveillance, the prevalence rates for NTDs decreased from 1.89 to.95 cases per 1000 live births and fetal deaths. The prevalence decrease is explained primarily by a decrease in cases of spina bifida. Isolated NTDs accounted for 297/360 (82%) NTDs and 63/360 (18%) had at least 1 other structural anomaly. Females predominated among isolated NTDs but the sex distribution was equal among NTD cases with other anomalies. Prevalence rates for whites (1.48 cases per 1000 live births and fetal deaths) were higher than rates for blacks (.87 cases per 1000 live births and fetal deaths). There were no NTD recurrences in 113 subsequent pregnancies to mothers of infants with isolated NTDs who took periconceptional folic acid. The rate of periconceptional folic acid use among women of childbearing years increased from 8% to 35% during the 6-year project period. CONCLUSION: The prevalence of NTDs in a high-risk region has declined coincident with the increased periconceptional use of folic acid supplements among women of childbearing age.neural tube defects, high-risk region, birth defects, folic acid, spina bifida, anencephaly, encephalocele. PMID- 11015509 TI - Pilot study of treatment with whole body hypothermia for neonatal encephalopathy. AB - BACKGROUND: There is extensive experimental evidence to support the investigation of treatment with mild hypothermia after birth asphyxia. However, clinical studies have been delayed by the difficulty in predicting long-term outcome very soon after birth and by concern about adverse effects of hypothermia. OBJECTIVES: The objectives of this study were to determine whether it is feasible to select infants with a bad neurological prognosis and to begin hypothermic therapy within 6 hours of birth, and to observe the effect of this therapy on relevant physiologic variables. METHODS: Sixteen newborn infants with clinical features of birth asphyxia (median cord blood pH: 6.74; range: 6.58-7.08) were assessed by amplitude integrated electroencephalography (aEEG), and mild whole body hypothermia was instituted within 6 hours of birth in the 10 infants with an aEEG prognostic of a bad outcome. Rectal temperature was maintained at 33.2 +/- (standard deviation).6 degrees C for 48 hours. Rectal and tympanic membrane temperature, blood pressure, heart rate, blood gases, blood lactate, full blood count, blood electrolytes, high and low shear rate viscosity, and coagulation studies were monitored during and after cooling. A preliminary assessment of neurological outcome was made by repeated magnetic resonance imaging (MRI) and neurological examination. RESULTS: All infants selected to receive hypothermia developed convulsions and a severe encephalopathy. During 48 hours of hypothermia infants had prolonged metabolic acidosis (median pH: 7.30; base excess: -6.3 mmol x L(-1), a high blood lactate (median lactate: 5.3 mmol x L(-1)) and low blood potassium levels (median value: 3.9 mmol x L(-1)) x Hypothermia was associated with lower heart rate and higher mean blood pressure. However, these changes did not seem to be clinically relevant and no significant complication of hypothermia was encountered. Blood viscosity and coagulation studies were similar during and after cooling. Unusual MRI findings were noted in 3 infants: transverse sinus thrombosis with subsequent small cerebellar infarct; probable thrombosis in the straight sinus; and hemorrhagic cerebral infarction. Six of the 10 cooled infants had minor abnormalities only or normal follow-up neurological examination; 3 infants died and 1 had major abnormalities. None of the 6 infants with a normal aEEG developed severe neonatal encephalopathy or neurological sequel. CONCLUSIONS: After birth asphyxia infants can be objectively selected by aEEG and hypothermia started within 6 hours of birth in infants at high risk of developing severe neonatal encephalopathy. Prolonged mild hypothermia to 33 degrees C to 34 degrees C is associated with minor physiologic abnormalities. Further studies of both the safety and efficacy of mild hypothermia, including further neuroimaging studies, are warranted. PMID- 11015510 TI - Pneumococcal mastoiditis in children. AB - OBJECTIVE: To determine the impact of antibiotic resistance on the frequency, clinical features, and management/outcome of mastoiditis attributable to Streptococcus pneumoniae. DESIGN: Retrospective review of the medical records of children with mastoiditis caused by S pneumoniae from September 1993 through December 1998. PATIENTS: Infants and children with pneumococcal mastoiditis cared for at 8 children's hospitals in the United States. RESULTS: Thirty-four children with pneumococcal mastoiditis were identified. The median age of the children was 12 months (range: 2 months-12.5 years); 28 (82%) were 100 mL/kg/day were tolerated (EI) or at 61 days of life (late enteral iron supplementation [LI]). Nutritional iron status was assessed: 1) at birth, 2) at 61 days of life, 3) when the infants reached a weight of 1.6 times birth weight, and 4) before blood was transfused at a hematocrit of <.25. ID was defined by any one of the following criteria: ferritin, <12 microg/L; transferrin saturation, <17%; or increase of absolute reticulocyte counts by >50% one week after the onset of enteral iron supplementation. Restrictive red cell transfusion guidelines were followed and all transfusions were documented. Erythropoietin was not administered. The primary outcome variables were: 1) ferritin at 61 days and 2) the number of infants with ID. RESULTS: Ferritin at 61 days was not different between the groups. Infants in the LI group were more often iron-deficient (26/65 vs 10/68) and received more blood transfusions after day 14 of life. No adverse effects of EI were noted. CONCLUSIONS: EI is feasible and probably safe in infants with birth weight <1301 g. EI may reduce the incidence of ID and the number of late blood transfusions. ID may occur in very low birth weight infants despite early supplementation with iron and should be considered in the case of progressive anemia.preterm infant, iron supplementation, iron deficiency, blood transfusion. PMID- 11015512 TI - Venous and arterial hematologic profiles of very low birth weight infants. European Multicenter rhEPO Study Group. AB - OBJECTIVE: In very low birth weight (VLBW) infants, diagnostic and therapeutic decisions depend on hematologic values. As few data are available, we studied the course during the first 6 weeks of life. DESIGN: Four prospective longitudinal cohort studies were retrospectively combined assessing hematologic profiles of 562 VLBW infants. For characterization of red blood cells and iron, infants receiving erythropoietin were excluded. For characterization of white blood cells and platelets, infants receiving antibiotics were excluded. RESULTS: The third (3rd)/median/97th percentiles on day 3 were as follows: hemoglobin: 11.0/15.6/19.8 g/dL; hematocrit: 35/47/60%; red blood cells: 3.2/4.2/5.3 x 10(12)/L; reticulocytes:. 6/7.1/27.8%; platelets: 58/203/430 x 10(9)/L; white blood cells: 3. 6/9.5/38.3 x 10(9)/L; neutrophils:.7/4.7/25.3 x 10(9)/L; ferritin: 27/140/504 ng/mL; iron:.8/7.5/26.7 micromol/L; transferrin saturation: 2.6/22.7/79.8%. Transferrin saturation was <24% in 51%, ferritin concentration <100 ng/mL in 32%, and platelets <150 x 10(9)/L in 29% of this population. The steady decrease of red cell parameters was mitigated by transfusions. Neutrophils decreased steadily, and were <1.75 x 10(9)/L in 35% at 6 weeks. CONCLUSIONS: Iron indices and platelet counts on day 3 and neutrophil counts at 2 to 6 weeks of age are lower than previously assumed in VLBW infants and lower than in larger prematures. PMID- 11015513 TI - When to suspect fungal infection in neonates: A clinical comparison of Candida albicans and Candida parapsilosis fungemia with coagulase-negative staphylococcal bacteremia. AB - OBJECTIVES: To determine the epidemiology of candidemia in our neonatal intensive care unit; to compare risk factors, clinical presentation, and outcomes for neonates infected with Candida albicans, Candida parapsilosis, and coagulase negative staphylococcus (CoNS); and to suggest a rational approach to empiric antifungal therapy of neonates at risk for nosocomial infection. DESIGN: Retrospective chart review of all neonatal intensive care unit patients with systemic candidiasis or CoNS infection between January 1, 1995 and July 31, 1998 at Duke University Medical Center. RESULTS: Fifty-one patients were reviewed. Nine of 19 patients infected with C parapsilosis and 5 of 15 patients infected with C albicans died of fungemia. Seventeen neonates had >2 positive cultures for CoNS obtained within 96 hours and 1 died. There was no statistically significant difference in birth weight, gestational age, or age at diagnosis between patient groups; however, candidemic patients had a sevenfold higher mortality rate. Before diagnosis, candidemic patients had greater exposure to systemic steroids, antibiotics, and catecholamine infusions. Of the 51 patients, 32 received third generation cephalosporins in the 2 weeks before diagnosis and 19 did not. Twenty nine of the 32 who were treated with third-generation cephalosporins subsequently developed candidemia, while candidemia occurred in only 5 of 19 patients who were not treated with cephalosporins. At the time of diagnosis, candidemic patients were more likely to have required mechanical ventilation and were less likely to be tolerating enteral feeding. Multivariate clustered logistic regression analysis revealed that candidemic patients had more exposure to third-generation cephalosporins. Once the clinician was notified of a positive blood culture for Candida, patients infected with C parapsilosis retained their central catheters longer than patients infected with C albicans. CONCLUSIONS: In this retrospective review, we were able to identify aspects of the clinical presentation and medication history that may be helpful in differentiating between candidemia and CoNS bacteremia. Those key features may be used by clinicians to initiate empiric amphotericin B therapy in premature neonates at risk for nosocomial infections. Prolonged use of third-generation cephalosporins was strongly associated with candidemia. There was no statistically significant difference in the morbidity and mortality between patients infected with C parapsilosis and those infected with C albicans. Observed delays in removal of the central venous catheter may have contributed to finding a mortality rate from C parapsilosis that was higher than was previously reported. PMID- 11015514 TI - Different beta-casein fractions in Icelandic versus Scandinavian cow's milk may influence diabetogenicity of cow's milk in infancy and explain low incidence of insulin-dependent diabetes mellitus in Iceland. AB - OBJECTIVES: To compare children with insulin-dependent diabetes mellitus (IDDM) with controls in Iceland regarding their consumption of cow's milk in infancy, and to investigate the beta-casein fractions in Scandinavian and Icelandic cow's milk. The A1 variant of beta-casein has been shown to be diabetogenic in animal studies, and suggestions have been made that the B variant of beta-casein acts similarly. Differences in the relative proportions of beta-casein fractions might explain the lower incidence of IDDM in Iceland than in Scandinavia. METHODS: A retrospective case-control study on IDDM patients and matching controls was performed in Iceland to compare their diets in infancy. Fifty-five children with IDDM born in Iceland over a 16-year period and randomly collected controls (n = 165) were recruited to the study. Mothers of the children answered questions on breastfeeding habits and on when cow's milk products were introduced. Samples of cow's milk from randomly selected milk batches from the largest consumption areas in Iceland and Scandinavia were collected. The milk samples were freeze-dried and their beta-casein fractions were analyzed using capillary electrophoresis. RESULTS: No significant difference was found between IDDM patients and controls in the frequency and duration of breastfeeding or the first introduction of cow's milk products. The analyses of milk samples showed that the percentage of the A1 and B variants of beta-casein in Icelandic milk was significantly lower than in the milk from the Scandinavian countries. CONCLUSIONS: Cow's milk consumption in infancy is not related to IDDM in Iceland. The lower fraction of A1 and B beta caseins in Icelandic cow's milk may explain why there is a lower incidence of IDDM in Iceland than in Scandinavia. PMID- 11015515 TI - Otitis media in childhood in relation to preschool language and school readiness skills among black children. AB - OBJECTIVE: To examine whether otitis media with effusion (OME) and associated hearing loss (HL) during the first 5 years of life were related to children's language skills during the preschool years and to school readiness skills at entry to kindergarten. METHODS: In a prospective study, the ears of 85 black children primarily from low-income families and recruited from community-based childcare programs were repeatedly examined from 6 months to 5 years of age for the presence of OME and from 6 months to 4 years of age for HL when well and ill with OME. Assessments were made annually of the children's child-rearing environments at home and in childcare, and children's language skills between 3 and 5 years of age and readiness skills in literacy and math were evaluated at entry into kindergarten. RESULTS: Children had either bilateral or unilateral OME approximately 30.4% and HL 19.6% of the observation time. OME and associated HL were significantly positively correlated with some measures of expressive language at 3 and 4 years of age; however, these direct relationships were no longer significant when the child's gender, socioeconomic status, maternal educational level, and the responsiveness and support of the home and childcare environments were also considered. Further, both OME and HL were moderately correlated with school readiness skills at entry to school, with children having more OME scoring lower in verbal math problems and with children with more HL scoring lower in math and recognizing incomplete words. These associations continued to remain significant even after partialing out the child and family background factors. CONCLUSIONS: There was not a significant relationship between children's early OME history or HL and language skills during the preschool years. However, children with more frequent OME had lower scores on school readiness measures. These associations were moderate in degree, however, and the home environment was more strongly related to academic outcomes than was OME or HL. These results should be interpreted cautiously when generalizing to other populations. PMID- 11015516 TI - Large congenital melanocytic nevi and the risk for development of malignant melanoma and neurocutaneous melanocytosis. AB - OBJECTIVE: To determine the risk for developing malignant melanoma and neurocutaneous melanocytosis (NCM) in patients with large congenital melanocytic nevi. DESIGN: Follow-up data suitable for calculations were available on 160 patients in the New York University Registry of Large Congenital Melanocytic Nevi who had been free of known melanomas or NCM when entered into the Registry. The cumulative 5-year life-table risks for developing melanoma and NCM were calculated. The relative risk for developing melanoma, using a control general population reference group, was determined. RESULTS: The 160 patients (median age at entry: 14 months) were followed prospectively for an average of 5.5 years. Three extracutaneous melanomas developed: 2 were in the central nervous system (CNS) and 1 was retroperitoneal. The 5-year cumulative life-table risk for developing melanoma was 2.3% (95% confidence interval [CI]:.8-6.6) and the relative risk was 101 (95% CI: 21-296). No melanoma occurred within a large congenital melanocytic nevus. Four patients developed manifest NCM, 2 with CNS melanomas. The 5-year cumulative life-table risk for developing NCM was 2.5% (95% CI:.8-7.2). Ten patients were excluded from the calculations because of preexisting disease on entry into the Registry: 5 with manifest NCM and 5 with melanomas (3 in large congenital melanocytic nevi, 1 in nonnevus skin, and 1 unknown primary). CONCLUSIONS: Patients with large congenital melanocytic nevi are at increased risk for developing melanomas. There is also a significant increased risk for developing NCM. The high incidence of CNS involvement may influence decisions concerning treatment of the large congenital melanocytic nevi. PMID- 11015517 TI - Prospective evaluation of propofol anesthesia in the pediatric intensive care unit for elective oncology procedures in ambulatory and hospitalized children. AB - OBJECTIVES: To evaluate our experience with propofol anesthesia delivered by pediatric intensivists in the pediatric intensive care unit (PICU) to facilitate elective oncology procedures in children performed by pediatric oncologists. METHODS: Elective oncology procedures performed with propofol anesthesia in our multidisciplinary, university-affiliated PICU were prospectively evaluated over a 7-month period. Ambulatory and hospitalized children were prescheduled for their procedure, underwent a medical evaluation, and met fasting requirements before the start of anesthesia. Continuous cardiorespiratory and neurologic monitoring was performed by a pediatric intensivist and a PICU nurse, while the procedure was performed by a pediatric oncologist. Propofol was delivered in intermittent boluses to achieve the desired level of anesthesia. Information studied included patient demographics, procedures performed, induction and total doses of propofol used, the duration of the different phases of the patient's PICU stay, the occurrence of side effects, the need for therapeutic interventions, and the incidence of recall of the procedure. RESULTS: Fifty procedures in 28 children (mean age: 7.5 +/- 4.3 years) were evaluated. Sixty-one percent of patients had established diagnoses. Fifty-four percent of procedures were lumbar puncture with intrathecal chemotherapy administration and 26% of procedures were bone marrow aspirations with biopsy. Induction propofol doses were 2. 0 +/-.8 mg/kg for ambulatory and hospitalized patients, while total propofol doses were 6.6 +/- 2.3 mg/kg and 7.9 +/- 2.4 mg/kg for ambulatory and hospitalized patients, respectively. Induction time was 1.5 +/-.7 minutes, recovery time was 23.4 +/- 11.5 minutes, and total PICU time was 88.8 +/- 27.7 minutes. Transient decreases in systolic blood pressure less than the fifth percentile for age occurred in 64% of procedures, with a mean decrease of 25% +/- 10%. Intravenous fluids were administered in 31% of these cases. Hypotension was more common in ambulatory patients but was not predicted by propofol dose, anesthesia time, or age. Partial airway obstruction was noted in 12% of procedures while apnea requiring bag-valve mask ventilation occurred in 2% of procedures. Neither was associated with age, propofol dose, or the duration of anesthesia. All procedures were successfully completed and there were no incidences of recall of the procedure. CONCLUSIONS: Propofol anesthesia is effective in achieving patient comfort and amnesia, while optimizing conditions for elective oncology procedures in children. Although transient hypotension and respiratory depression may occur, propofol anesthesia seems to be safe to use for these procedures in the PICU setting. Recovery from anesthesia was rapid and total stay was brief. Under the proper conditions, propofol anesthesia delivered by pediatric intensivists in the PICU is a reasonable option available to facilitate invasive oncology procedures in children. PMID- 11015518 TI - Depressive symptoms and cigarette smoking among teens. AB - CONTEXT: Although several risk factors for tobacco use have been implicated in the development of depression, smoking progression has typically been viewed as a consequence of depression. The results of limited longitudinal studies are controversial. OBJECTIVE: To assess the nature and direction of the relationship between cigarette smoking and depression among teens. DESIGN: Prospective analysis of baseline and 1-year follow-up data from the National Longitudinal Study of Adolescent Health. SETTING: In-home teen and parent interviews. PARTICIPANTS: Two samples were identified. For the first sample, 8704 adolescents who were not depressed at baseline based on CES-D scores were identified for analyses of the effects of cigarette smoking on development of high depressive symptomatology. Baseline smoking status, which could vary in this group, was the predictor of interest in these analyses. For the second sample, 6947 teens who had not smoked cigarettes in the 30 days before the baseline survey (noncurrent smokers) were identified for analyses on the effect of high depressive symptoms on subsequent moderate to heavy cigarette use at 1 year of follow-up. Baseline high depressive symptomatology based on CES-D score was the predictor of interest in this sample. MAIN OUTCOME MEASURES: Among the nondepressed, developing high depressive symptoms at 1 year of follow-up. Among noncurrent smokers, smoking at least 1 pack per week at 1 year of follow-up. RESULTS: For the nondepressed, multivariate modeling revealed that current cigarette smoking was the strongest predictor of developing high depressive symptoms in all models (final model odds ratio [OR]: 3.90; 95% confidence interval [CI]: 1.85,8.20). For noncurrent smokers, although in bivariate analyses, baseline high depressive symptoms increased the risk of heavy smoking nearly threefold, multivariate modeling revealed that baseline high depressive symptoms were not predictive of heavy smoking when controlling for other determinants of smoking in teens. Previous experimentation with smoking was the strongest predictor of becoming a heavy smoker (OR: 3.04; 95% CI: 1.93,4.88). CONCLUSIONS: In contrast to common dictum, depression does not seem to be an antecedent to heavy cigarette use among teens. However, current cigarette use is a powerful determinant of developing high depressive symptoms. PMID- 11015519 TI - Racial differences in access to the kidney transplant waiting list for children and adolescents with end-stage renal disease. AB - CONTEXT: Renal transplantation is the treatment of choice for pediatric patients with end-stage renal disease (ESRD). Black patients wait longer for kidney transplants than do white patients. OBJECTIVE: To determine whether the increased time to transplantation for black pediatric patients is attributable not only to a shortage of suitable donor organs, but also to racial differences in the time from a child's first treatment for ESRD until activation on the cadaveric kidney transplant waitlist. DESIGN: National longitudinal cohort study. SETTING: US Medicare-eligible, pediatric ESRD population. PATIENTS: Children and adolescents 3000 g at birth and with HCs lower than the fifth percentile had Hg in meconium. CONCLUSION: The higher prevalence and levels of Hg in the fetal compartments reflect the ease of placental transfer with fetal trapping. Hg determinations in the mothers' blood underestimate the degree and extent of fetal exposure. There is a significant difference in each compartment's ability to reflect Hg exposure of the fetus. A small HC may be associated with the presence of Hg in meconium. Hg in meconium should be measured in addition to cord blood to determine the load of fetal Hg. PMID- 11015524 TI - Patterns of alcohol and drug use in adolescents can be predicted by parental substance use disorders. AB - OBJECTIVE: To examine the specificity of risk for alcohol or drug abuse or dependence (substance use disorders [SUDs]) in offspring exposed to particular subtypes of parental SUDs. METHODS: The original sample was derived from 2 groups of index children: 140 attention-deficit/hyperactivity disorder (ADHD) probands and 120 non-ADHD comparison probands. These groups had 174 and 129 biological siblings and 279 and 240 parents, respectively. RESULTS: Independent of familial risk, exposure to parental SUDs predicted SUDs in the offspring. Controlling for duration of exposure, we found that adolescence was a critical developmental period for exposure to parental SUDs. Because all our analyses controlled for social class, ADHD status, and parental lifetime history of SUDs, these results show that exposure to parental SUDs predicts offspring SUDs independently of these risk factors. CONCLUSIONS: These results support the critical importance of familial environmental risk factors for the development of SUDs in youth in general and particularly in those at high risk for these disorders. These results highlight adolescence as a critical period for the deleterious effects of exposure to parental SUDs, supporting the need to develop preventive and early intervention strategies targeted at adolescents at high risk for SUDs. PMID- 11015523 TI - Teacher-assessed behavior of children prenatally exposed to cocaine. AB - OBJECTIVE: Prenatal cocaine exposure has been associated with alterations in neonatal behavior and more recently a dose-response relationship has been identified. However, few data are available to address the long-term behavioral effects of prenatal exposures in humans. The specific aim of this report is to evaluate the school-age behavior of children prenatally exposed to cocaine. METHODS: All black non-human immunodeficiency virus-positive participants in a larger pregnancy outcomes study who delivered singleton live born infants between September 1, 1989 and August 31, 1991 were eligible for study participation. Staff members of the larger study extensively screened study participants during pregnancy for cocaine, alcohol, cigarettes, and other illicit drugs. Prenatal drug exposure was defined by maternal history elicited by structured interviews with maternal and infant drug testing as clinically indicated. Cocaine exposure was considered positive if either history or laboratory results were positive. Six years later, 665 families were contacted; 94% agreed to participate. The child, primary caretaker (parent), and, when available, the biologic mothers were tested in our research facilities. Permission was elicited to obtain blinded teacher assessments of child behavior with the Achenbach Teacher's Report Form (TRF). Drug use since the child's birth was assessed by trained researchers using a structured interview. RESULTS: Complete laboratory and teacher data were available for 499 parent-child dyads, with a final sample size for all analyses of 471 (201 cocaine-exposed) after the elimination of mentally retarded subjects. A comparison of relative Externalizing (Aggressive, Delinquent) to Internalizing (Anxious/Depressed, Withdrawn, Somatic Complaints) behaviors of the offspring was computed for the TRF by taking the difference between the 2 subscales to create an Externalizing-Internalizing Difference (T. M. Achenbach, personal communication, 1998). Univariate comparisons revealed that boys were significantly more likely to score in the clinically significant range on total TRF, Externalizing-Internalizing, and Aggressive Behaviors than were girls. Children prenatally exposed to cocaine had higher Externalizing-Internalizing Differences compared with controls but did not have significantly higher scores on any of the other TRF variables. Additionally, boys prenatally exposed to cocaine were twice as likely as controls to have clinically significant scores for externalizing (25% vs 13%) and delinquent behavior (22% vs 11%). Gender, prenatal exposures (cocaine and alcohol), and postnatal risk factors (custody changes, current drug use in the home, child's report of violence exposure) were all related to problem behaviors. Even after controlling for gender, other prenatal substance exposures, and home environment variables, cocaine-exposed children had higher Externalizing-Internalizing Difference scores. Prenatal exposure to alcohol was associated with higher total score, increased attention problems, and more delinquent behaviors. Prenatal exposure to cigarettes was not significantly related to the total TRF score or any of the TRF subscales. Postnatal factors associated with problem behaviors included both changes in custody status and current drug use in the home. Change in custody status of the cocaine-exposed children, but not of the controls, was related to higher total scores on the TRF and more externalizing and aggressive behaviors. Current drug use in the home was associated with higher scores on the externalizing and aggressive subscales. CONCLUSIONS: Results of this study suggest gender-specific behavioral effects related to prenatal cocaine exposure. Prenatal alcohol exposure also had a significant impact on the TRF. Postnatal exposures, including current drug use in the home and the child's report of violence exposure, had an independent effect on teacher-assessed child behavioral problems. (ABSTRACT TRUNCATE PMID- 11015525 TI - Year 2000 position statement: principles and guidelines for early hearing detection and intervention programs. Joint Committee on Infant Hearing, American Academy of Audiology, American Academy of Pediatrics, American Speech-Language Hearing Association, and Directors of Speech and Hearing Programs in State Health and Welfare Agencies. PMID- 11015526 TI - The unnecessary epidemic of folic acid-preventable spina bifida and anencephaly. PMID- 11015527 TI - Commentary on cerebral intravascular oxygenation correlates with mean arterial pressure in critically ill premature infants. PMID- 11015528 TI - Verve and Jolt: deadly new Internet drugs. AB - As regulatory agencies have increased restrictions on the sale and marketing of gamma-hydroxybutyrate (GHB), they have been frustrated by the appearance of precursor molecules such as gamma-butyrolactone (GBL) that have become widely available over the Internet. These dangerous precursors are vigorously marketed to adolescents and young adults as dietary supplements that increase muscle mass and enhance sexual performance with seductive names such as Verve and Jolt, both easily recognizable teen icons. We present the case of an adolescent who ingested both of these GBL products 2 weeks apart, resulting in life-threatening respiratory depression and emergent intubation on both occasions. The GBL toxidrome, necessary acute interventions, and public health implications are reviewed. We urge all health care providers to report similar cases immediately to the FDA MedWatch system. Gamma-butyrolactone, gamma-hydroxybutyrate, respiratory insufficiency, central nervous system depressants, substance abuse. PMID- 11015529 TI - Role of naloxone in newborn resuscitation. AB - OBJECTIVE: Because of questions about the basis for the use of naloxone in resuscitation of the newborn, we wished to evaluate the use of naloxone at our institution and an affiliated hospital. METHODOLOGY: Evaluation of the actual use of naloxone at a university hospital and a community hospital: we document naloxone use by daily survey for a month in one; in the other, we perform a retrospective record review of 1 year's use. RESULTS: The university hospital had 240 births during February, 1998. Naloxone was given twice: once, 7 minutes before delivery to a woman at term who had received opiates about 2 hours previously; and once, intramuscularly, to a premature infant for apnea, before being intubated. The community hospital had 2044 births during fiscal 1998. Twenty-six neonates were identified as having received naloxone. Of the 26, 13 received naloxone without needing ventilatory support; all 13 with respiratory depression had a predisposing perinatal complication. CONCLUSION: The use of naloxone in practice may not conform to the American Academy of Pediatrics' guidelines for use in resuscitation of the newborn. The use of naloxone in resuscitation of the newborn should be reevaluated. PMID- 11015530 TI - Respiratory syncytial virus infection in patients with phagocyte defects. AB - Patients with phagocyte defects frequently develop bacterial or fungal pneumonias, but they are not considered to be at increased risk for viral infections. We describe 3 patients with known phagocyte immunodeficiencies who developed lower respiratory tract infections (LRTI) caused by respiratory syncytial virus (RSV). All 3 patients had dense pneumonias as indicated by computed tomography scan of the lungs and RSV was recovered. We conclude that RSV can present as a dense pneumonia in patients with phagocyte defects. Along with common pathogens causing LRTI, RSV should be considered in the differential diagnosis. Viral cultures as well as rapid antigen detection assays for respiratory viruses should be included in the evaluation of LRTI in patients with phagocyte defects. respiratory syncytial virus, phagocyte, immunodeficiency, pneumonia. PMID- 11015532 TI - "Blue balls": A diagnostic consideration in testiculoscrotal pain in young adults: A case report and discussion. PMID- 11015531 TI - Visual field constriction in children with epilepsy on vigabatrin treatment. AB - Vigabatrin is considered the drug of choice for infantile spasms and simple and complex partial epilepsy in childhood. Its mechanism of action relies on the irreversible inhibition of gamma-aminobutyric acid (GABA) transaminase. Since June 1997 several articles have been published reporting visual field constriction in adult patients on vigabatrin therapy. Recently, 7 pediatric patients, 1 on vigabatrin monotherapy and 6 on add-on therapy with visual field constriction have been described. We have observed 30 pediatric patients with epilepsy (14 boys and 16 girls), ages ranging from 4 to 20 years (mean: 11 years and 2 months) treated with vigabatrin for infantile spasms, simple and complex partial epilepsy, who had never complained of ophthalmologic disturbances. Twenty one patients underwent complete routine ophthalmologic examination (fundus oculi, visual acuity, intraocular pressure, and visual field tests); 9 children (<6 years old) underwent only fundus examination, because collaboration was lacking. We report on 4 children showing constriction of visual field, prevailing in nasal hemifield. In 1 child, visual abnormalities were stable even 10 months after vigabatrin discontinuation, while in another a greater improvement was observed 5 months after discontinuation. The possible mechanisms have been discussed and the cone dysfunction, connected with GABA augmentation in the outer retina, has been outlined. We suggest a possible protocol to control visual abnormalities in epileptic children. PMID- 11015533 TI - Isolated large third-trimester intracranial cyst on fetal ultrasound: fact or fiction? AB - OBJECTIVE: To distinguish the fact from artifact of an isolated, large, intracranial cyst on prenatal sonography (PSG). BACKGROUND: The use of PSG is rapidly increasing with most obstetric ultrasounds occurring in general community settings like small hospitals and clinics with personnel who have variable training, experience, and interest levels. In contrast, most PSG articles and books are produced in large subspecialty centers with concentrated referral bases plus both highly-trained and experienced personnel. DESIGN/METHODS: We report a series of 2 normal newborn patients who had a large prenatal unilateral intracranial cyst diagnosed by PSG in the 10 years between July of 1989 and 1999 at a rural community hospital. The newborns had imaging studies at birth and their neurodevelopmental progress was followed for several years. Textbook, bibliography and computerized Medline (1966-present) searches including prenatal ultrasound, observer variation, diagnostic errors, reproducibility of results, sensitivity and specificity, accuracy, central nervous system, false-positive, prenatal diagnosis, and brain were examined starting in August 1996 for reports. RESULTS: There were 4079 obstetric ultrasounds performed in 3.5 years, January 1996 through July 1999 at this rural community facility. This rate extrapolates to a total of 11 654 obstetric ultrasounds over the 10-year study period in which the 2 cases of intracranial cyst artifact occurred. Thus, the incidence of 2 intracranial cyst artifacts was estimated as 2/11 654 PSG, a .0002% false positive rate. CONCLUSIONS: This is the first report of the occurrence of PSG artifacts in a community facility. Artifact is a real problem and needs to be specified in differential diagnoses. There are ways to decrease sonographic artifact-or at least to recognize it-so our estimates at a community hospital for its occurrence are presented with the relevant technical and ethical issues. None of these issues have been previously reported in the pediatric literature. Our false-positive rate for large intracranial cyst compares favorably with other reports. Our estimate may inflate our denominator by reporting scans rather than the number of fetuses scanned, and our numerator may miss cases that moved from the community. Confusion differentiating PSG artifact from reality often occurs when interpreting static or frozen real-time images. The signs that sonogram images may be artifacts include defects that: extend outside the fetal body; change shape, size and echogenecity with different scan planes; are not seen on all examinations; and are isolated in an otherwise normal fetus. Failure to offer quality PSG in clinical settings where it is available restricts access of pregnant women to the diagnosis of fetal anomalies, and therefore restricts access to the options of pregnancy termination, fetal therapy like fetal surgery, and delivery options of timing, setting, and mode. We suggest a multidisciplinary approach to prenatal abnormalities like isolated third trimester unilateral intracranial cyst in both primary and tertiary care settings aids interpretation followed by expectant conservative management without elaborate, risky, or terminal interventions. PMID- 11015534 TI - Successful long-term peritoneal dialysis in a very low birth weight infant with renal failure secondary to feto-fetal transfusion syndrome. PMID- 11015535 TI - Use of a peak flow meter for positive feedback in psychogenic cough. PMID- 11015536 TI - The role of the pediatrician in transitioning children and adolescents with developmental disabilities and chronic illnesses from school to work or college. American Academy of Pediatrics. Committee on Children With Disabilities. AB - The role of the pediatrician in transitioning children with disabilities and chronic illnesses from school to work or college is to provide anticipatory guidance and to promote self-advocacy and self-determination. Knowledge of the provisions of the key federal laws affecting vocational education is essential for the pediatrician's successful advocacy for patients. PMID- 11015537 TI - Children in pickup trucks. American Academy of Pediatrics. Committee on Injury and Poison Prevention. AB - Pickup trucks have become increasingly popular in the United States. A recent study found that in crashes involving fatalities, cargo area passengers were 3 times more likely to die than were occupants in the cab. Compared with restrained cab occupants, the risk of death for those in the cargo area was 8 times higher. Furthermore, the increased use of extended-cab pickup trucks and air bag-equipped front passenger compartments creates concerns about the safe transport of children. The most effective preventive strategies are the legislative prohibition of travel in the cargo area and requirements for age-appropriate restraint use and seat selection in the cab. Parents should select vehicles that are appropriate for the safe transportation needs of the family. Physicians have an important role in counseling families and advocating public policy measures to reduce the number of deaths and injuries to occupants of pickup trucks. PMID- 11015538 TI - Insurance coverage of mental health and substance abuse services for children and adolescents: A consensus statement. American Academy of Pediatrics. PMID- 11015539 TI - Primary care treatment of pediatric psychosocial problems: A study from pediatric research in office settings and ambulatory sentinel practice network. AB - OBJECTIVE: Psychosocial problems cause much of the morbidity among children, and their frequency of presentation in primary care is growing. How is primary care treatment of children's psychosocial problems affected by child symptoms, physician training, practice structure, insurance, physician/patient relationship, and family demographics? DESIGN: Questionnaire study of treatment of psychosocial problems during office visits by children. SETTINGS: At total of 401 primary care offices from 44 US states, Puerto Rico, and Canada. PATIENTS: From 21 150 children seen in office visits, we selected children with an identified psychosocial problem but who were not already receiving specialty mental health services (n = 2618 children). OUTCOME MEASURES: Clinicians' decisions to counsel families, to refer children to mental health specialists, or to prescribe medication. RESULTS: The treatment choices of primary care clinicians (PCCs) were generally independent of patients' demographics or insurance status. Clinicians' training, beliefs about mental health, and practice structure had no effect on treatment choices. However, clinicians seeing their own patients were more likely to prescribe medications for attention problems. The clinician's perception about whether the parent agreed with the treatment choice was important for every treatment modality. Counseling and referral were more common and medication was less common when a problem was newly recognized at the visit. CONCLUSIONS: Structural factors such as practice type, insurance coverage, and physician training were less important for treatment than were process factors, such as whether the visit was a psychosocial problem visit, whether the problem was newly or previously recognized, and whether the family and clinician were familiar with each other and in accord about treatment. PMID- 11015540 TI - Time for a new paradigm in pediatric medical education: teaching neonatal resuscitation in a simulated delivery room environment. AB - OBJECTIVES: Acquisition and maintenance of the skills necessary for successful resuscitation of the neonate are typically accomplished by a combination of completion of standardized training courses using textbooks, videotape, and manikins together with active participation in the resuscitation of human neonates in the real delivery room. We developed a simulation-based training program in neonatal resuscitation (NeoSim) to bridge the gap between textbook and real life and to assess trainee satisfaction with the elements of this program. METHODS: Thirty-eight subjects (physicians and nurses) participated in 1 of 9 full-day NeoSim programs combining didactic instruction with active, hands-on participation in intensive scenarios involving life-like neonatal and maternal manikins and real medical equipment. Subjects were asked to complete an extensive evaluation of all elements of the program on its conclusion. RESULTS: The subjects expressed high levels of satisfaction with nearly all aspects of this novel program. Responses to open-ended questions were especially enthusiastic in describing the realistic nature of simulation-based training. The major limitation of the program was the lack of fidelity of the neonatal manikin to a human neonate. CONCLUSION: Realistic simulation-based training in neonatal resuscitation is possible using current technology, is well received by trainees, and offers benefits not inherent in traditional paradigms of medical education. PMID- 11015541 TI - Television commercial violence during nonviolent programming: the 1998 major league baseball playoffs. AB - OBJECTIVES: To identify the frequency of violent commercials during the major league baseball playoffs in 1998 and to compare it with the 1996 playoffs. DESIGN AND SETTING: All 4 World Series games televised on the Fox Television Network (Fox), all 6 National League Championship Series (NLCS) televised by Fox, and 5 of 6 American League Championship Series (ALCS) playoff games televised by the National Broadcasting Company (NBC) were videotaped in October 1998. The results were compared with a similar study that analyzed the 1996 playoffs. RESULTS: Forty-four commercials (11.0 per game) that included violent interactions were noted during the World Series, 53 violent commercials (8.8 per game) during the NLCS, and 40 (8.0 per game) during the ALCS for a total of 137. Of these 137 violent commercials, 78 contained at least 1 violent act, 126 contained at least 1 violent threat, and 12 contained evidence of at least 1 violent consequence. One hundred five of the 137 violent commercials (76.6%) were television promotions and 23 (16.8%) were for big-screen movie promotions. Twenty-three of 35 big-screen movie promotions (65.7%) were violent. Ten of the 137 violent commercials (7.3%) contained obvious blood, of which 9 were televised by NBC. There were, however, an additional 20 commercials televised by Fox that contained visible blood if the videotape was slowed or paused. Sixty-three of the 137 violent commercials (46.0%) used fire and explosions, of which 53 were televised by Fox and 10 by NBC. Guns were involved in 86 of 137 violent commercials (62.8%), with Fox accounting for 64 (6.4 per game) and NBC accounting for 22 (4.4 per game). Knives were used in 1 violent commercial on both Fox and NBC. Comparisons that can be made to 1996 include the following: 1) violent commercials during the World Series (both televised by Fox) increased from 10.1 to 11.0 per game; and 2) violent commercials during the ALCS (both televised by NBC) increased from 6.0 to 8.0 per game. CONCLUSION: Commercials during the 1998 major league playoffs were similar in violent content, compared with 1996. The violent commercials continue to consist primarily of promotions for television programs and big-screen movies. It continues to be counterintuitive to find such commercials in family-oriented programming and makes it difficult for parents to avoid exposing their children to this form of violence. PMID- 11015542 TI - Environmental tobacco smoke exposure among urban children. AB - BACKGROUND: Environmental tobacco smoke (ETS) exposure is a well-known health hazard for children. Poor urban children may be at increased risk of exposure from nonparental sources of tobacco smoke. The unique problems of ETS exposure in urban children have previously not been well-defined. OBJECTIVES: This study was conducted: 1) to identify the sources of ETS exposure for children attending a hospital-based pediatric resident practice; and 2) to examine caregivers' perceptions of the pediatrician's role and action in addressing ETS exposure. DESIGN: Questionnaires were administered to a convenience sample of 174 caregivers of children visiting a children's hospital-based resident practice. A structured face-to-face interview was used. RESULTS: All caregivers agreed to participate in the study. Most of the caregivers interviewed were mothers (87.9%). Although only 14% of caregivers were smokers, 49.4% reported caring for at least 1 smoke-exposed child. Most smoke exposure occurred in the home; smoking is allowed in the home of 75% of the children who live with a smoker (48/64). Thirty-four percent of the caregivers who reported no smoking in the home reported having a child who spends time in homes where other people smoke. Most exposure outside the home occurred at a grandparent's home. A ban on smoking in the home did not eliminate a child's risk of residential smoke exposure but did predict whether smoking occurred in the same room as the child. All caregivers reported believing that tobacco smoke is harmful. Approximately one half of caregivers who smoke (54.2%) would like smoking cessation advice from the child's pediatrician. Interest in receiving advice on helping others quit smoking was also high (51.3%) among nonsmoking caregivers who cared for a smoke-exposed child. CONCLUSIONS: Nonparental sources of ETS exposure are reported to be common in urban children. Children may be exposed to tobacco smoke even in homes where smoking is banned. Many caregivers would like help from pediatricians in reducing children's exposure to tobacco smoke. PMID- 11015543 TI - Long-term effect of dust control on blood lead concentrations. AB - BACKGROUND: Dust control is recommended to prevent children's exposure to residential lead hazards, but the long-term effect of dust control on children's exposure to environmental lead is unknown. OBJECTIVE: To determine the effect of dust control on children's exposure to lead, as measured by blood lead concentration at 48 months of age. DESIGN: A randomized, controlled trial. Setting. Rochester, New York. PARTICIPANTS: A total of 275 urban children were randomized at 6 months of age; 189 (69%) were available for the 48-month follow up blood test. Intervention. Children and their families were randomly assigned to an intervention group that received cleaning equipment and up to 8 visits by a trained lead hazard control advisor or to a control group. The intervention was terminated when the children were 24 months of age. OUTCOME MEASURES: Geometric mean blood lead concentration and prevalence of elevated blood lead concentration (ie, >/=10 microg/dL, >/=15 microg/dL, and >/=20 microg/dL), by group assignment. RESULTS: For children with 48-month blood tests, baseline geometric mean blood lead concentrations were 2.8 microg/dL (95% confidence interval [CI]: 2.6, 3.0); there were no significant differences in baseline characteristics or lead exposure by group assignment. At 48 months of age, the geometric mean blood lead was 5.9 microg/dL (95% CI: 5.3, 6.7) for the intervention group and 6.1 microg/dL (95% CI: 5.5,6.9) for the control group. The percentage of children with a 48 month blood lead >/=10 microg/dL, >/=15 microg/dL, and >/=20 microg/dL was 19% versus 19%, 2% versus 9%, and 1% versus 2% in the intervention and control groups, respectively. CONCLUSIONS: We conclude that dust control, as performed by families and in the absence of lead hazard controls to reduce ongoing contamination from lead-based paint, was not effective in preventing children's exposure to residential lead hazards. PMID- 11015544 TI - Relations of parental obesity status to physical activity and fitness of prepubertal girls. AB - OBJECTIVE: To determine whether physical activity and fitness differ among normal weight-for-height, multiethnic, prepubertal girls whose parents are lean, obese, or both. SETTING: Conducted in Houston, Texas. STUDY PARTICIPANTS: Normal-weight white, black, and Hispanic prepubertal girls (mean age +/- standard deviation: 8.5 +/-.4 years) participated in this study. Girls were recruited according to parental leanness or obesity defined as follows: girls with 2 lean parents (LN group; n = 30); girls with 2 obese parents (OB group; n = 27); and girls with 1 lean and 1 obese parent (LNOB group; n = 44). Intervention. Each child wore a heart rate monitor for two 24-hour periods, underwent a treadmill exercise test, answered an activity questionnaire, and completed energy expenditure measurements by basal calorimetry and doubly labeled water. OUTCOME MEASURES: The amount of time spent above 125% and 150% of basal heart rate (BHR) was calculated during each 24-hour period (n = 84). Fitness (n = 97), habitual physical activity (n = 101), and physical activity level (PAL = total energy expenditure/basal metabolic rate; n = 101) were also measured. RESULTS: The times spent above 125% and 150% of BHR were similar among LN, LNOB, and OB groups. Black girls spent less time than did white girls on the weekend above 125% BHR and above 150% BHR. No significant familial or ethnic differences in peak oxygen consumption or habitual physical activity were observed. PALs were as follows: LN = 1.6 +/-.21; LNOB = 1.64 +/-.27; OB = 1.58 +/-. 20; white = 1.63 +/-.23; black = 1.58 +/-.24; and Hispanic = 1.65 +/-.25. PAL was related to time spent above 125% BHR and 150% BHR (r =.31,.39). CONCLUSIONS: Physical activity and fitness of normal-weight prepubertal girls predisposed to obesity did not differ from girls without a family history of obesity. obesity, children, energy metabolism, exercise, heart rate monitoring. PMID- 11015545 TI - Aerobic fitness, not energy expenditure, influences subsequent increase in adiposity in black and white children. AB - BACKGROUND: Low levels of energy expenditure and aerobic fitness have been hypothesized to be risk factors for obesity. Longitudinal studies to determine whether energy expenditure influences weight gain in whites have provided conflicting results. To date, no studies have examined this relationship in blacks or whether aerobic fitness influences weight gain in white or black children. METHODS: One hundred fifteen children, 72 white (55 girls and 17 boys) and 43 black (24 girls and 19 boys) were recruited for this study. Aerobic fitness, resting, total, and activity-related energy expenditure and body composition were measured at baseline. The children returned annually for 3 to 5 repeated measures of body composition. The influence of the initial measures of energy expenditure and fitness on the subsequent rate of increase in adiposity was examined, adjusting for initial body composition, age, ethnicity, gender, and Tanner stage. Because 20 children did not attain maximum oxygen consumption, the sample size for the combined analysis was 95. RESULTS: Initial fat mass was the main predictor of increasing adiposity in this cohort of children, with greater initial fat predicting a higher rate of increase of adiposity. There was also a significant negative relationship between aerobic fitness and the rate of increasing adiposity (F(1,82) = 3.92). With every increase of.1 L/minute of fitness, there was a decrease of.081 kg fat per kg of lean mass gained. None of the measures of energy expenditure significantly predicted increasing adiposity in white or black children. CONCLUSIONS: Initial fat mass was the dominant factor influencing increasing adiposity; however, aerobic fitness was also a significant independent predictor of increasing adiposity in this cohort of children. Resting, total, or activity-related energy expenditure did not predict increasing adiposity. It seems that aerobic fitness may be more important than absolute energy expenditure in the development of obesity in white or black children. energy expenditure, fitness, longitudinal, obesity. PMID- 11015546 TI - Infant immunization with acellular pertussis vaccines in the United States: assessment of the first two years' data from the Vaccine Adverse Event Reporting System (VAERS). AB - OBJECTIVE: To evaluate the safety of infant immunization with acellular pertussis vaccines in the United States. BACKGROUND: The US Food and Drug Administration approved the first acellular pertussis vaccine for use in infants in the United States on July 31, 1996. OUTCOME MEASURES: Adverse events in the United States after infant immunization with pertussis-containing vaccines, representing temporal (but not necessarily causal) associations between vaccinations and adverse events. DATA SOURCE: Reports to the Vaccine Adverse Event Reporting System (VAERS), a passive national surveillance system. DESIGN: Reports concerning infant immunization against pertussis between January 1, 1995 (when whole-cell vaccine was in exclusive use) and June 30, 1998 (when acellular vaccine was in predominant use) were analyzed, if the reports were entered into the VAERS database by November 30, 1998. RESULTS: During the study, there were 285 reports involving death, 971 nonfatal serious reports, and 4514 less serious reports after immunization with any pertussis-containing vaccine. For 1995 there were 2071 reports; in 1996 there were 1894 reports; in 1997 there were 1314 reports, and in the first half of 1998 there were 491 reports. Diphtheria-tetanus pertussis vaccine (DTP) was cited in 1939 reports, diphtheria-tetanus-whole-cell pertussis-Haemophilus influenzae type b vaccine (DTPH) in 2918 reports, and diphtheria-tetanus-acellular pertussis vaccine (DTaP) in 913 reports. The annual number of deaths during the study was 85 in 1995, 82 in 1996, 77 in 1997, and 41 in the first half of 1998. The annual number of reported events categorized as nonfatal serious (defined as events involving initial hospitalization, prolongation of hospitalization, life-threatening illness, or permanent disability) to VAERS for all pertussis-containing vaccines declined: 334 in 1995, 311 in 1996, 233 in 1997, and 93 in the first half of 1998. Similarly, the annual number of less serious reports to VAERS for pertussis-containing vaccines declined: 1652 in 1995, 1501 in 1996, 1004 in 1997, and 357 in the first half of 1998. A comparison of the adverse event profiles (proportional distributions) for DTaP, DTP, and DTPH, as well as an analysis of specific adverse events considered in a 1991 Institute of Medicine report on the safety of diphtheria-tetanus pertussis vaccine, did not identify any new, clear safety concerns. CONCLUSIONS: These findings reflect the administration of millions of doses of acellular pertussis vaccine and are reassuring with regard to the safety of marketed acellular pertussis vaccines. VAERS data, although subject to the limitations of passive surveillance, support the prelicensure data with regard to the safety of the US-licensed acellular pertussis vaccines that we evaluated. PMID- 11015547 TI - Hypotonic-hyporesponsive episodes reported to the Vaccine Adverse Event Reporting System (VAERS), 1996-1998. AB - BACKGROUND: A hypotonic-hyporesponsive episode (HHE) is the sudden onset of hypotonia, hyporesponsiveness, and pallor or cyanosis that occurs within 48 hours after childhood immunizations. This syndrome has been primarily associated with pertussis-containing vaccines administered to children <2 years of age, and has been estimated to occur once every 1750 diphtheria-tetanus-pertussis (DTwP) vaccinations. Previous studies of HHE were limited by small numbers of cases and, sometimes, by limited details of the event. OBJECTIVES: To characterize a large number of HHE cases reported to the Vaccine Adverse Event Reporting System (VAERS), to assist clinicians in identifying HHE, and to assist researchers in investigating the risk factors, cause, and pathogenesis of this syndrome. METHODS: More than 40,000 VAERS reports received between 1996 and 1998 were screened for HHE by a computer algorithm and reviewed, and a telephone follow-up questionnaire was administered to the witness of HHE. RESULTS: There were 215 HHE cases, all nonfatal. The median age of onset of HHE was 4.0 months (range: 1.1 107 months). Over half of the reports (53%) concerned females. The median birth weight was 3. 36 kg (range: 1.27-4.96 kg); 4.7% had a birth weight <2500 g. The median interval between vaccination and HHE was 210 minutes (range: 1 minute-2 days). Among children with HHE who were <24 months of age, the episode occurred within 5 minutes in only 8.5%, compared with 66.7% of children with HHE >24 months of age. There were no relevant findings regarding family medical history or the mothers' gestational history. Nearly all of the children (98.6%) returned to their prevaccination state according to the telephone questionnaire; median time to return was 6 hours (range: 1 minute- 4 months). The 3 children reported as not returning to their prevaccination state all had VAERS reports submitted after they developed conditions (autism, complex partial epilepsy, and developmental delays with infantile spasms) that are not known to be causally associated with immunization. The vast majority of children (93%) with HHE received a pertussis-containing vaccine, either diphtheria-tetanus-acellular pertussis (DTaP, 28%), DTwP (11%), or diphtheria-tetanus-pertussis-Haemophilus influenzae type b (DTwP-HIB, 61%). During the HHE episode, 90.1% of the children had pallor and 49% had cyanosis. Because of the HHE event, 6.8% of children had had all vaccines withheld as of the date of the interview. Of the remainder, 66.5% of children have had 1 or more subsequent vaccinations or vaccine components withheld, and 26.7% have not had any subsequent vaccinations withheld. Only 1 child was reported to have had a repeat episode of HHE, occurring after hepatitis B vaccination. From 1996 to 1998, the number of HHE reports decreased from 99 to 38, when the predominant pertussis vaccine administered to infants changed from whole-cell to acellular. CONCLUSION: This study represents the largest published case series of children with HHE and supports the generally benign, self-limited, nonrecurrent nature of this syndrome. Although HHE has been less frequently reported to VAERS after increased use of DTaP, HHE does occur after the administration of DTaP and other nonpertussis-containing vaccines. Although most parents and pediatricians withheld the pertussis component of subsequent vaccinations, many did not, with no reported adverse events occurring in the children after the subsequent immunizations. Restricting the definition of HHE to a more narrow age range (eg, <2 years of age) is also proposed because most of the older children probably experienced vasovagal syncope rather than HHE within 5 minutes of immunization. PMID- 11015548 TI - Interferon-alpha and ribavirin in treating children and young adults with chronic hepatitis C after malignancy. AB - OBJECTIVE: Chronic hepatitis C is a major long-term problem for children who survive cancer. Interferon (IFN)-alpha has been shown to be effective in treating patients with chronic hepatitis C; however, the rate of sustained response is low. Combining IFN-alpha and ribavirin (RBV) has been shown to significantly improve the response in adult patients with chronic hepatitis C. The aim of this pilot study was to evaluate the efficacy and safety of a combined virostatic treatment with IFN-alpha and RBV in a small cohort of children and adolescents with chronic hepatitis C and previous malignancy. METHODS: Twelve patients with a history of a hematooncologic disease (median follow-up: 13.5 years; range: 7-14.7 years) and chronic hepatitis C were treated with recombinant IFN-alpha-2a (6 megaunits/m(2) body surface area, 3 times a week, subcutaneously) combined with RBV (15 mg/kg body weight/day, orally) for 12 months. They were tested monthly for blood counts and liver function, and for serum virus concentrations (hepatitis C virus RNA by polymerase chain reaction) every 3 months. RESULTS: At the end of the treatment, hepatitis C virus RNA could not be detected in the serum of 8 of the 12 patients; 2 of these patients relapsed soon after therapy withdrawal, whereas 6 patients maintained in sustained virologic and biochemical remission (follow-up: 12 months). Treatment-induced toxicity was moderate and reversible with influenza-like symptoms and a decrease in blood counts in all 12 patients, alopecia in 5 of the 12, hemolysis in 4 of the 12, and weight loss of >10% in 2 of the 12. CONCLUSIONS: As demonstrated in adults with chronic hepatitis C, treatment with IFN-alpha and RBV also seems to be an effective and safe therapeutic option for children and adolescents with chronic hepatitis C after malignancy. PMID- 11015549 TI - Sources of infection among persons with acute hepatitis A and no identified risk factors during a sustained community-wide outbreak. AB - CONTEXT: Hepatitis A is a common vaccine-preventable disease in the United States. Most cases occur during community-wide outbreaks, which can be difficult to control. Many case-patients have no identified source. OBJECTIVE: To identify foodborne and household sources of hepatitis A during a community-wide outbreak. DESIGN: Serologic and descriptive survey. SETTING: Salt Lake County, Utah. PARTICIPANTS: A total of 355 household contacts of 170 persons reported with hepatitis A during May 1996 to December 1996, who had no identified source of infection; and 730 food handlers working in establishments where case-patients had eaten. MAIN OUTCOME MEASURE: Prevalence of immunoglobulin M antibodies to hepatitis A virus (IgM anti-HAV) among household and food service contacts. RESULTS: Overall, 70 household contacts (20%) were IgM anti-HAV-positive, including 52% of children 3 to 5 years old and 30% of children <3 years old. In multivariate analysis, the presence of a child <3 years old (odds ratio [OR]: 8.8; 95% confidence limit [CL]: 2.1,36) and a delay of >/=14 days between illness onset and reporting (OR: 7. 9; 95% CL: 1.7,38) were associated with household transmission. Of 18 clusters of infections linked by transmission between households, 13 (72%) involved unrecognized infection among children <6 years old. No food handlers were IgM anti-HAV-positive. CONCLUSION: During a community-wide outbreak, HAV infection among children was common, was frequently unrecognized, and may have been an important source of transmission within and between households. Transmission from commercial food establishments was uncommon. Ongoing vaccination of children may prevent future outbreaks. PMID- 11015550 TI - Late-onset neutropenia in very low birth weight infants. AB - BACKGROUND: Neutropenia, defined as absolute neutrophil count (ANC) <1500/mm(3), affects 6% to 58% of premature infants in the first week of life. This early onset neutropenia in premature infants has previously been correlated with sepsis, maternal hypertension, severe asphyxia, and periventricular hemorrhage. Late-onset neutropenia, defined as ANC <1500/mm(3) at a postnatal age of >/=3 weeks, has not been previously reported. OBJECTIVES: The purposes of this study were to determine the prevalence of late-onset neutropenia in very low birth weight (VLBW) infants and to examine the factors that may be associated with this phenomenon. METHODS: A weekly complete blood cell count (CBC) was performed routinely in all premature infants with birth weight , His(468), and Glu354)) and the SEDCLY motif places human macrophage CEH in the family of carboxylesterases. A greater than 20-fold increase in CEH activity was observed when COS-1 and COS-7 cells were transiently transfected with an eukaryotic expression vector, pcDNA3.1/V5/His-TOPO, containing the cDNA for human macrophage CEH. Using this full-length cDNA as a probe, a 2.2-kb transcript was identified by Northern blot analysis of total RNA from human peripheral blood and THP -1 macrophages. Overexpression of human macrophage CEH resulted in an impairment of upregulation of low-density lipoprotein (LDL) receptor mRNA in Chinese hamster ovary (CHO-K1) cells grown in cholesterol-deficient environment. These data identify the human macrophage CEH, demonstrate its expression in human peripheral blood macrophage and human macrophage cell line, THP-1, and suggest its role in the intracellular cholesteryl ester metabolism. PMID- 11015576 TI - A novel mutation in FGFR-3 disrupts a putative N-glycosylation site and results in hypochondroplasia. AB - Fibroblast growth factor receptor 3 (FGFR3) is a glycoprotein that belongs to the family of tyrosine kinase receptors. Specific mutations in the FGFR3 gene are associated with autosomal dominant human skeletal disorders such as hypochondroplasia, achondroplasia, and thanatophoric dysplasia. Hypochondroplasia (HCH), the mildest form of this group of short-limbed dwarfism disorders, results in approximately 60% of cases from a mutation in the intracellular FGFR3-tyrosine kinase domain. The remaining cases may either be caused by defects in other FGFR gene regions or other yet unidentified genes. We describe a novel HCH mutation, the first found outside the common mutation hot spot of this condition. This point mutation, an N328I exchange in the extracellular Ig domain III of the receptor, seems to be unique as it affects a putative N-glycosylation site that is conserved between different FGFRs and species. The amino acid exchange itself most probably has no impact on the three-dimensional structure of the receptor domain, suggesting that the phenotype is the result of altered receptor glycosylation and its pathophysiological consequences. PMID- 11015577 TI - Inflammation influences vascular remodeling through AT2 receptor expression and signaling. AB - The AT(2) receptor, which exerts growth inhibitory effects in cell culture, is present scantily in the adult vasculature but is reexpressed after vascular injury. To examine the in vivo role of this receptor in vascular diseases, we developed a mouse model of vascular remodeling and compared the responses in wild type (Agtr2(+)) and AT(2) receptor knockout (Agtr2(-)) mice. Polyethylene cuff placement on the femoral artery led to the vascular expression of cytokines, the transcriptional factor interferon regulatory factor-1 (IRF-1), and both the AT(1) and AT(2) receptors. Although the expressions of IRF-1 and AT(1) receptor were induced to comparable levels in both the Agtr2(+) and Agtr2(-) mice, the neointimal lesion size and the smooth muscle cell proliferation were twice greater in the Agtr2(-) than in the Agtr2(+) mouse. Correlated with this difference, AT(2) receptor expression was induced predominantly in the smooth muscle cells of Agtr2(+) mouse. These results demonstrate that the AT(2) receptor plays an important role in nonocclusive inflammatory injury by mediating the effects of inflammation on vascular smooth muscle growth inhibition. PMID- 11015578 TI - nNOS and eNOS modulate cGMP formation and vascular response in contracting fast twitch skeletal muscle. AB - Nitric oxide (NO) from Ca(2+)-dependent neuronal nitric oxide synthase (nNOS) in skeletal muscle fibers may modulate vascular tone by a cGMP-dependent pathway similar to NO derived from NOS in endothelial cells (eNOS). In isolated fast twitch extensor digitorum longus (EDL) muscles from control mice, cGMP formation increased approximately 166% with electrical stimulation (30 Hz, 15 s). cGMP levels were not altered in slow-twitch soleus muscles. The NOS inhibitor N(omega) nitro-l-arginine abolished the contraction-induced increase in cGMP content in EDL muscles, and the NO donor sodium nitroprusside (SNP) increased cGMP content approximately 167% in noncontracting EDL muscles. SNP treatment but not electrical stimulation increased cGMP formation in muscles from nNOS(-/-) mice. cGMP formation in control and stimulated EDL muscles from eNOS(-/-) mice was less than that obtained with similarly treated muscles from control mice. Arteriolar relaxation in contracting fast-twitch mouse cremaster muscle was attenuated in muscles from mice lacking either nNOS or eNOS. These findings suggest that increases in cGMP and NO-dependent vascular relaxation in contracting fast-twitch skeletal muscle may require both nNOS and eNOS. PMID- 11015579 TI - Assessment of cellular profile and lung function with repeated bronchoalveolar lavage in individual mice. AB - In this study, we sought to develop procedures that would enable repeated bronchoalveolar lavage (BAL) in individual mice on multiple occasions. To achieve this objective, we first developed the procedures that would allow individual mice to survive a whole lung lavage, and then tested whether, on subsequent days, there was an effect of this initial BAL on the cell profile, lung permeability, and baseline respiratory function. Our results demonstrate that the repeated lavage procedure can be readily carried out in individual mice of different strains on multiple occasions. The lavage procedure itself results in immediate increases in respiratory system resistance and concomitant decreases in compliance, but these parameters return to prelavage values by the 2nd or 3rd day postlavage. Lavage also induces variable increases in inflammatory cells depending on the strain used. However, in all three strains examined here (A/J, BALB/c, and C3H/HeJ), inflammatory cell numbers returned to baseline values within 3 days after an initial lavage procedure. The ability to perform repeated BAL in individual mice should prove to be an extremely useful tool in a variety of functional genomic studies in the lung. PMID- 11015580 TI - Reverse genetics in zebrafish. AB - The zebrafish has become a popular model system for the study of vertebrate developmental biology because of its numerous strengths as a molecular genetic and embryological system. To determine the requirement for specific genes during embryogenesis, it is necessary to generate organisms carrying loss-of-function mutations. This can be accomplished in zebrafish through a reverse genetic approach. This review discusses the current techniques for generating mutations in known genes in zebrafish. These techniques include the generation of chromosomal deletions and the subsequent identification of complementation groups within deletions through noncomplementation assays. In addition, this review will discuss methods currently being evaluated that may improve the methods for finding mutations in a known sequence, including screening for randomly induced small deletions within genes and screening for randomly induced point mutations within specific genes. PMID- 11015581 TI - Realizing the potential of zebrafish as a model for human disease. AB - The value of the zebrafish (Danio rerio) as a model for human disease has been substantiated by a number of recently published papers. Several zebrafish mutants with "human" diseases have been found, spanning a variety of human pathologies. These successful studies utilizing the zebrafish have been made possible by the development of key reagents such as YAC, PAC, and BAC libraries, as well as radiation hybrid panels. With the further establishment of new tools and access to the newly generated resources, the zebrafish is poised to serve as a novel model for human disease. PMID- 11015582 TI - Chips around the world: proceedings from the Nature Genetics microarray meeting. PMID- 11015583 TI - Mass spectrometry and proteomics. PMID- 11015584 TI - Targeting the Hprt locus in mice reveals differential regulation of Tie2 gene expression in the endothelium. AB - To study the in vivo expression of the murine Tie2 gene, we have targeted the hypoxanthine phosphoribosyltransferase (Hprt) gene locus to generate two single copy transgenic mice: T1, containing the 2,100-bp Tie2 promoter upstream from the beta-galactosidase (LacZ) gene, and T5, which also included an enhancing element originating from the first intron of the Tie2 gene. Comparing T1 and T5 embryos at day E10.5 revealed differential endothelial cell-specific expression of LacZ, whereas colocalization analyses showed that the expression was confined to endothelial cells. Moderate reporter gene activity was observed in the brain and kidney of T1 adults, whereas extensive LacZ gene expression was seen in the vasculature of most organs of the T5 adults. This study demonstrates the feasibility of targeting the Hprt locus with endothelial cell-specific sequences to analyze the spatial-temporal expression of transgenes. Of particular importance is the observation that the analysis of a single transgene copy in a defined locus allows for an accurate and rapid comparison of transcriptional activity among regulatory DNA sequences. PMID- 11015585 TI - Targeting of human eNOS promoter to the Hprt locus of mice leads to tissue restricted transgene expression. AB - Phenotypic heterogeneity of the endothelium arises from cell type-specific differences in gene expression. An understanding of the mechanisms that underlie differential gene expression would provide important insight into the molecular basis of vascular diversity. In standard transgenic assays, multiple copies of heterologous DNA cassettes are randomly integrated into the mouse genome, resulting in significant line-to-line variation in expression. To overcome these limitations, we have targeted a single copy of a transgene that contains 1,600 bp of the human endothelial nitric oxide synthase (eNOS) promoter coupled to the LacZ reporter gene to the X-linked hypoxanthine phosphoribosyltransferase (Hprt) locus of mice by homologous recombination. The transgene was inserted in either of the orientations relative to that of the Hprt gene. In mice derived from multiple embryonic stem (ES) cell clones, the expression pattern was limited to a subset of endothelial cells, cardiomyocytes, and vascular smooth muscle cells. These findings suggest that Hprt locus targeting is a feasible tool for studying endothelial cell-restricted gene regulation. PMID- 11015586 TI - Encroaching genomics: adapting large-scale science to small academic laboratories. AB - The process of conducting biological research is undergoing a profound metamorphosis due to the technological innovations and torrent of information resulting from the execution of multiple species genome projects. The further tasks of mapping polymorphisms and characterizing genome-wide protein-protein interaction (the characterization of the proteome) will continue to garner resources, talent, and public attention. Although some elements of these whole genome size projects can only be addressed by large research groups, consortia, or industry, the impact of these projects has already begun to transform the process of research in many small laboratories. Although the impact of this transformation is generally positive, laboratories engaged in types of research destined to be dominated by the efforts of a genomic consortium may be negatively impacted if they cannot rapidly adjust strategies in the face of new large-scale competition. The focus of this report is to outline a series of strategies that have been productively utilized by a number of small academic laboratories that have attempted to integrate such genomic resources into research plans with the goal of developing novel physiological insights. PMID- 11015588 TI - Regulation of leptin by agouti. AB - Dominant mutations at the mouse Agouti locus lead to ectopic expression of the Agouti gene and exhibit diabetes, obesity, and yellow coat color. Obese yellow mice are hyperinsulinemic and hyperleptinemic, and we hypothesized that Agouti directly induces leptin secretion. Accordingly, we used transgenic mice expressing agouti in adipocytes (under the control of aP2 promoter, aP212) to examine changes in leptin levels. Agouti expression in adipose tissue did not significantly alter food intake, weight gain, fat pad weight, or insulinemia; however, the transgenic mice were hyperglycemic. We demonstrated that plasma leptin levels are approximately twofold higher in aP212 transgenic mice compared with their respective controls, whereas ubiquitous expression of agouti (under the control of beta-actin promoter, BAP20) led to a sixfold increase in leptin. Insulin treatment of aP212 mice increased adipocyte leptin content without affecting plasma leptin levels. These findings were further confirmed in vitro in 3T3-L1 adipocytes treated with recombinant Agouti protein and/or insulin. Agouti but not insulin significantly increased leptin secretion, indicating that insulin enhances leptin synthesis but not secretion while Agouti increases both leptin synthesis and secretion. This increased leptin synthesis and secretion was due to increased leptin mRNA levels by Agouti. Interestingly, agouti regulation of leptin was not mediated by melanocortin receptor 4, previously implicated in agouti regulation of food intake. These results suggest that increased leptin secretion by agouti may serve to limit agouti-induced obesity, independent of melanocortin receptor antagonism, and indicate that interaction between obesity genes may play a key role in obesity. PMID- 11015587 TI - Gene expression profile in mouse myocardium after ischemia. AB - This study was designed to elaborate a molecular profile of expressed genes during ischemic injury to the mouse heart after surgical constriction of the left coronary artery without reperfusion. A mouse cDNA array containing 588 known genes was used to compare gene expression in heart RNA after 24-h ischemia with control tissue. Alterations in gene expression on the array were supported by relative reverse transcription-polymerase chain reaction analysis after timed periods of ischemia. Decreased levels of the cell cycle regulator p18ink4 and the oxidative responsive gene glutathione S-transferase were accompanied by an upregulation of the genes associated with cardiac muscle development, alpha myosin heavy chain and fetal myosin alkali light chain. Other stress responses elicited by cardiac injury included an induction of Egr-1 and Egr-3 transcription factors, as well as the apoptotic regulator Bax. Altogether, these findings indicate that expression of genes associated with a fetal transcription program may be involved with the post ischemic remodeling process in heart ventricles. PMID- 11015589 TI - Genetically defined risk of salt sensitivity in an intercross of Brown Norway and Dahl S rats. AB - A genetic segregation analysis was performed to identify genes that cosegregate with arterial blood pressure traits reflective of salt sensitivity. A population of 113 F2 male rats was derived from an intercross of inbred SS/JrHsd/Mcw (Dahl salt-sensitive) and BN/SsN/Mcw (Brown Norway) rats. Rats were maintained on an 8% salt diet from the age of 9 to 13 wk, and arterial pressure was measured for 3 h daily during the 4th wk of high salt intake in unanesthetized rats using implanted arterial catheters. At the end of the 3rd day of high-salt pressure recordings, the arterial pressure response to salt depletion was determined 1.5 days following treatment with Lasix and a low-sodium (0. 4%) diet. A genome-wide scan using 265 polymorphic simple sequence length polymorphism (SSLP) markers found that seven arterial pressure phenotypes determined at different times and circumstances, and representing two distinct indexes of salt sensitivity, mapped to the same region of rat chromosome 18. The trait of salt sensitivity was strongly influenced by the presence of SS alleles in this region of chromosome 18, and those rats which were homozygote SS/SS exhibited a significantly greater reduction of mean arterial pressure following sodium depletion (29 +/- 2 mmHg) than homozygote BN/BN (17 +/- 3 mmHg) or heterozygotic (22 +/- 2 mmHg) rats. This region of rat chromosome 18 corresponds to the long arm of human chromosome 5 and a region of human chromosome 18 that has been linked to hypertension in humans. Given the unlikely chance of these different blood pressure traits mapping to the same region, we believe these data provide evidence that this region of rat chromosome 18 plays an important role in salt-induced hypertension. PMID- 11015590 TI - Adeno-associated virus vector transduction of vascular smooth muscle cells in vivo. AB - Adeno-associated virus (AAV) vectors might offer solutions for restenosis and angiogenesis by transducing nondividing cells and providing long-term gene expression. We investigated the feasibility of vascular cell transduction by AAV vectors in an in vivo rabbit carotid artery model. Time course of gene expression, inflammatory reaction to the vector, and effects of varying viral titer, exposure time, and intraluminal pressures on gene expression were examined. Recombinant AAV vectors with an Rous sarcoma virus promoter and alkaline phosphatase reporter gene were injected intraluminally into transiently isolated carotid segments. Following transduction, gene expression increased significantly over 14 days and then remained stable to 28 days, the last time point examined. Medial vascular smooth muscle cells were the main cell type transduced even with an intact endothelial layer. Increasing the viral titer and intraluminal pressure both enhanced transduction efficiency to achieve a mean of 34 +/- 7% of the subintimal layer of smooth muscle cells expressing gene product. A mild inflammatory reaction, composed of T cells with only rare macrophages, with minimal intimal thickening was demonstrated in 40% of transduced vessels; inflammatory cells were not detected in sham-operated control arteries. These findings demonstrate that AAV is a promising vector for intravascular applications in coronary and peripheral vascular diseases. PMID- 11015591 TI - Inactivation of one copy of the mouse neurotrophin-3 gene induces cardiac sympathetic deficits. AB - Whether two copies of the neurotrophin-3 (NT3) gene are necessary for proper development of cardiac sympathetic innervation was investigated in mice carrying a targeted inactivation of the NT3 gene. Heterozygous (+/-) and null (-/-) mutant mice had fewer stellate ganglion neurons than did wild-type (+/+) mice at postnatal day 0 (P0 or birth), and this deficit was maintained between adult (P60) +/- and +/+ mice. The sympathetic innervation of the heart matured postnatally in +/+ and +/- mice. Tyrosine hydroxylase (TH)-positive axons were restricted largely to the epicardium at P0, were concentrated around large blood vessels in the myocardium at P21, and were present among cardiac myocytes at P60. Cardiac norepinephrine (NE) concentrations paralleled the growth of the sympathetic axons into the heart. NE concentrations were equivalent among +/+, +/ , and -/- mice at birth, but differences between +/- and +/+ mice increased with age. Adult +/- mice also exhibited lower resting heart rates and sympathetic tonus than +/+ mice. Thus deletion of one copy of the NT3 gene translates into anatomical, biochemical, and functional deficits in cardiac sympathetic innervation of postnatal mice, thereby indicating a gene-dosage effect for the NT3 gene. PMID- 11015592 TI - Angiotensin peptides acting at rostral ventrolateral medulla contribute to hypertension of TGR(mREN2)27 rats. AB - We have previously demonstrated that microinjections of the selective angiotensin (1-7) [ANG-(1-7)] antagonist, A-779, into the rostral ventrolateral medulla (RVLM) produces a significant fall in mean arterial pressure (MAP) and heart rate (HR) in both anesthetized and conscious rats. In contrast, microinjection of angiotensin II (ANG II) AT(1) receptor antagonists did not change MAP in anesthetized rats and produced dose-dependent increases in MAP when microinjected into the RVLM of conscious rats. In the present study, we evaluated whether endogenous ANG-(1-7) and ANG II acting at the RVLM contribute to the hypertension of transgenic rats harboring the mouse renin Ren-2 gene, TGR(mREN2)27. Unilateral microinjection of A-779 (0.1 nmol) produced a significant fall in MAP (-25 +/- 5 mmHg) and HR (-57 +/- 20 beats/min) of awake TGR rats. The hypotensive effect was greater than that observed in Sprague-Dawley (SD) rats (-9 +/- 2 mmHg). Microinjection of the AT(1) antagonist CV-11974 (0.2 nmol) produced a fall in MAP in TGR rats (-14 +/- 4 mmHg), contrasting with the pressor effect observed in SD rats (33 +/- 9 mmHg). These results indicate that endogenous ANG-(1-7) exerts a significant pressor action in the RVLM, contributing to the hypertension of TGR(mREN2)27 transgenic rats. The role of ANG II at the RVLM seems to be dependent on its endogenous level in this area. PMID- 11015593 TI - Comparison of human adult and fetal expression and identification of 535 housekeeping/maintenance genes. AB - Gene expression levels of about 7,000 genes were measured in 11 different human adult and fetal tissues using high-density oligonucleotide arrays to identify genes involved in cellular maintenance. The tissues share a set of 535 transcripts that are turned on early in fetal development and stay on throughout adulthood. Because our goal was to identify genes that are involved in maintaining cellular function in normal individuals, we minimized the effect of individual variation by screening mRNA pooled from many individuals. This information is useful for establishing average expression levels in normal individuals. Additionally, we identified transcripts uniquely expressed in each of the 11 tissues. PMID- 11015595 TI - A fuzzy logic approach to analyzing gene expression data. AB - We have developed a novel algorithm for analyzing gene expression data. This algorithm uses fuzzy logic to transform expression values into qualitative descriptors that can be evaluated by using a set of heuristic rules. In our tests we designed a model to find triplets of activators, repressors, and targets in a yeast gene expression data set. For the conditions tested, the predictions made by the algorithm agree well with experimental data in the literature. The algorithm can also assist in determining the function of uncharacterized proteins and is able to detect a substantially larger number of transcription factors than could be found at random. This technology extends current techniques such as clustering in that it allows the user to generate a connected network of genes using only expression data. PMID- 11015594 TI - The two isozymes of rat intestinal alkaline phosphatase are products of two distinct genes. AB - Rat intestinal alkaline phosphatases (IAP-I and -II) differ in primary structure, substrate specificity, tissue localization, and response to fat feeding. This study identifies two distinct genes ( approximately 5-6 kb) corresponding to each isozyme and containing 11 exons of nearly identical size. The exon-intron junctions are identical with those found in IAP genes from other species. The 1.7 and 1.2 bp of 5' flanking regions isolated from each gene, respectively, contain Sp1 and gut-enriched Kruppel-like factor (GKLF) binding sites, but otherwise show little identity. There is a potential CAAT-box 14 bp 5' to the transcriptional start site, 36 bp upstream from IAP-I, and a TATA-box 31 bp 5' to the transcriptional start site, 55 bp upstream from IAP-II. Transfection of these promoter regions (linked to luciferase as a reporter gene) into a kidney cell line, COS-7, produced the differential response to oleic acid expected from in vivo studies, i.e., threefold increase using the 5' flanking region of IAP-II, but not IAP-I. This response was not reproduced by 5,8,11,14-eicosatetraynoic acid (ETYA) or clofibrate, suggesting that peroxisome proliferator response elements are not involved. Isolation of the IAP-II gene will allow determination of the sequences responsible for dietary fat response in the enterocyte. PMID- 11015596 TI - Diet effects on weight gain and body composition in high growth (hg/hg) mice. AB - Nongenetic factors such as nutrition modulate the effects of genes responsible for overgrowth in animals. The goal of this study was to examine the importance of genotype x diet interactions on the effects of a major locus that regulates growth in the mouse. We have examined the phenotype of high growth (hg), a partially recessive autosomal locus that increases growth rate and mature body size. C57BL/6J (C57) and congenic C57BL/6J-hg/hg (HG) mice were fed three experimental diets differing in protein and energy content from 3 to 12 wk of age. HG mice grew faster and were, on average, 51% heavier than C57 at 12 wk of age. Feed intake was higher in HG mice but proportional to the increase in body weight. The magnitude of the differences in body size and composition between lines depended on the interaction between genotype and the protein/energy ratio of the diet. In C57, the diets modified the level of fatness without changing adult lean mass. However, in HG the diets differentially affected both linear growth and body composition. In general, HG had higher plasma levels of insulin like growth factor I at 3 and 12 wk than C57. Plasma insulin did not differ between lines, but leptin was higher for C57 mice fed a high-energy diet. These results show that the effects of hg on growth are modulated by diet composition. Therefore, this mutation could be a valuable model with which to study the genetic and nutritional aspects of overgrowth disorders. PMID- 11015597 TI - Identification of three human renin mRNA isoforms from alternative tissue specific transcriptional initiation. AB - We have reported that mice transgenic for 140- and 160-kb P1 phage artificial chromosomes (PACs) containing the human renin gene express the gene in a highly tissue-restricted and regulated manner. Herein, we demonstrate that the transgene is also expressed appropriately throughout development. In the course of this investigation, we identified the existence of three transcriptional isoforms of human renin mRNA derived from the utilization of alternative transcription start sites. The first isoform is the kidney-specific isoform, which utilizes the classic renin promoter. The second is a brain-specific isoform, which when previously identified in rats and mice was due to a transcription initiation site within intron A. However, the start site in the human gene resides approximately 1,325 bp upstream of the classic promoter and encodes a new exon 1 (termed exon 1b) that splices directly to exon 2. The third isoform is lung specific and is due to transcriptional initiation 79 bp directly upstream of exon 2, fusing additional DNA within intron A (termed exon 1c) directly to exon 2 without splicing. Importantly, the alternative first exons observed in the PAC transgenic mice were identical to those used to transcribe renin in human fetal kidney, brain, and lung, suggesting these sites are bona fide isoforms of human renin mRNA and not artifacts of transgenesis. Moreover, the subtle differences in tissue-specific transcriptional initiation observed in the renin gene of rats and humans can be faithfully and accurately emulated in a transgenic model. PMID- 11015598 TI - Blood pressure QTL that differentiate Dahl salt-sensitive and spontaneously hypertensive rats. AB - Our purpose was to define quantitative trait loci (QTL) for blood pressure that differ between two widely used hypertensive rat strains, the Dahl salt-sensitive (S) rat and the spontaneously hypertensive rat (SHR). A genome scan was done on an F(2) (S x SHR) population fed 8% NaCl for 4 wk. Three blood pressure QTL were detected, one on each of rat chromosomes (chr) 3, 8, and 9. For the chr 3 QTL the SHR allele increased blood pressure, and for chr 8 and 9 the S allele increased blood pressure. The QTL on chr 9 was exceptionally strong, having a LOD score of 7.3 and accounting for 30% of the phenotypic variance and a difference of 40 mmHg between homozygotes. A review of the literature in conjunction with the present data suggests that S and SHR are not different for the previously described prominent blood pressure QTL on chr 1, 2, 10, and 13. QTL for body weight on chr 4, 12, 18, and 20, each with an effect of about 30 g, were incidentally observed. PMID- 11015599 TI - Genetic variation in LMNA modulates plasma leptin and indices of obesity in aboriginal Canadians. AB - We previously showed that a rare mutation in LMNA, which encodes lamins A and C, underlies autosomal dominant Dunnigan-type familial partial lipodystrophy (FPLD). Because FPLD is an extreme example of genetically disturbed adipocyte differentiation, it is possible that common variation in LMNA is associated with obesity-related phenotypes. We therefore analyzed the relationships between the common LMNA 1908T/C single nucleotide polymorphism (SNP) and plasma leptin and anthropometric indices in 306 nondiabetic Canadian Oji-Cree. We found that subjects with the LMNA 1908T/1908T genotype had significantly higher plasma leptin than the subjects with either the 1908C/1908T or 1908C/1908C genotypes, after adjustment for age and sex. Physical indices of obesity, such as body mass index, percent body fat, and ratio of waist-to-hip circumference, were also higher among Oji-Cree subjects with the LMNA 1908T/1908T genotype than the subjects with either the 1908C/1908T or 1908C/1908C genotypes. The results suggest that common genetic variation in LMNA may be an important determinant of plasma leptin and obesity-related quantitative traits. PMID- 11015600 TI - ANP in regulation of arterial pressure and fluid-electrolyte balance: lessons from genetic mouse models. AB - The recent development of genetic mouse models presenting life-long alterations in expression of the genes for atrial natriuretic peptide (ANP) or its receptors (NPR-A, NPR-C) has uncovered a physiological role of this hormone in chronic blood pressure homeostasis. Transgenic mice overexpressing a transthyretin-ANP fusion gene are hypotensive relative to the nontransgenic littermates, whereas mice harboring functional disruptions of the ANP or NPR-A genes are hypertensive compared with their respective wild-type counterparts. The chronic hypotensive action of ANP is determined by vasodilation of the resistance vasculature, which is probably mediated by attenuation of vascular sympathetic tone at one or several prejunctional sites. Under conditions of normal dietary salt consumption, the hypotensive action of ANP is dissociated from the natriuretic activity of the hormone. However, during elevated dietary salt intake, ANP-mediated antagonism of the renin-angiotensin system is essential for maintenance of blood pressure constancy, inasmuch as the ANP gene "knockout" mice (ANP -/-) develop a salt sensitive component of hypertension in association with failure to adequately downregulate plasma renin activity. These findings imply that genetic deficiencies in ANP or natriuretic receptor activity may be underlying causative factors in the etiology of salt-sensitive variants of hypertensive disease and other sodium-retaining disorders, such as congestive heart failure and cirrhosis. PMID- 11015601 TI - Circadian rhythms in a nutshell. AB - Living organisms on this planet have adapted to the daily rotation of the earth on its axis. By means of endogenous circadian clocks that can be synchronized to the daily and seasonal changes in external time cues, most notably light and temperature, life forms anticipate environmental transitions, perform activities at biologically advantageous times during the day, and undergo characteristic seasonal responses. The effects of transmeridian flight and shift work are stark reminders that although modern technologies can create "cities that never sleep" we cannot escape the recalcitrance of endogenous clocks that regulate much of our physiology and behavior. Moreover, malfunctions in the human circadian timing system are implicated in several disorders, including chronic sleep disorders in the elderly, manic-depression, and seasonal affective disorders (SAD or winter depression). Recent progress in understanding the molecular mechanisms underlying circadian rhythms has been remarkable. In its most basic form, circadian clocks are comprised of a set of proteins that, by virtue of the design principles involved, generate a self-sustaining transcriptional-translational feedback loop with a free-running period of about 24 h. One or more of the clock components is acutely sensitive to light, resulting in an oscillator that can be synchronized to local time. This review provides an overview of the roles circadian clocks play in nature, how they might have arisen, human health concerns related to clock dysfunction, and mainly focuses on the clockworks found in Drosophila and mice, the two best studied animal model systems for understanding the biochemical and cellular bases of circadian rhythms. PMID- 11015602 TI - Role of intracellular calcium in human adipocyte differentiation. AB - Intracellular calcium ([Ca(2+)](i)) modulates adipocyte lipid metabolism and inhibits the early stages of murine adipogenesis. Consequently, we evaluated effects of increasing [Ca(2+)](i) in early and late stages of human adipocyte differentiation. Increasing [Ca(2+)](i) with either thapsigargin or A23187 at 0-1 h of differentiation markedly suppressed differentiation, with a 40-70% decrease in triglyceride accumulation and glycerol-3 phosphate dehydrogenase (GPDH) activity (P < 0.005). However, a 1-h pulse of either agent at 47-48 h only modestly inhibited differentiation. Sustained, mild stimulation of Ca(2+) influx with either agouti protein or 10 mM KCl-induced depolarization during 0-48 h of differentiation inhibited triglyceride accumulation and GPDH activity by 20-70% (P < 0.05) and markedly suppressed peroxisome proliferator-activated receptor gamma (PPARgamma) expression. These effects were reversed by Ca(2+) channel antagonism. In contrast, Ca(2+) pulses late in differentiation (71-72 h or 48-72 h) markedly increased these markers of differentiation. Thus increasing [Ca(2+)](i) appears to exert a biphasic regulatory role in human adipocyte differentiation, inhibiting the early stages while promoting the late stage of differentiation and lipid filling. PMID- 11015603 TI - Global expression profiling of yeast treated with an inhibitor of amino acid biosynthesis, sulfometuron methyl. AB - The expression pattern of 1,529 yeast genes in response to sulfometuron methyl (SM) was analyzed by DNA microarray technology. SM, a potent herbicide, inhibits acetolactate synthase, a branched-chain amino acid biosynthetic enzyme. Exposure of yeast cells to 0.2 microg/ml SM resulted in 40% growth inhibition, a Gcn4p mediated induction of genes involved in amino acid and cofactor biosynthesis, and starvation response. The accumulation of intermediates led to the induction of stress response genes and the repression of genes involved in carbohydrate metabolism, nucleotide biosynthesis, and sulfur assimilation. Extended exposure to SM led to a relaxation of the initial response and induction of sugar transporter and ergosterol biosynthetic genes, as well as repression of histone and lipid metabolic genes. Exposure to 5 microg/ml SM resulted in >98% growth inhibition and stimulated a similar initial expression change, but with no relaxation after extended exposure. Instead, more stress response and DNA damage repair genes become induced, suggesting a serious cellular consequence. Other salient features of metabolic regulation, such as the coordinated expression of cofactor biosynthetic genes with amino acid biosynthetic ones, were evident from our data. A potential link between SM sensitivity and ergosterol metabolism was uncovered by expression profiling and confirmed by genetic analysis. PMID- 11015604 TI - A novel, sensitive detection system for high-density microarrays using dendrimer technology. AB - To improve signal detection on cDNA microarrays, we adapted a fluorescent oligonucleotide dendrimeric signal amplification system to microarray technology. This signal detection method requires 16-fold less RNA for probe synthesis, does not depend on the incorporation of fluorescent dNTPs into a reverse transcription reaction, generates a high signal-to-background ratio, and can be used to allow for multichannel detection on a single chip. Furthermore, since the dendrimers can be detected individually, it may be possible, by employing dendrimer-binding standards, to calculate the numbers of bound cDNAs can be estimated. These features make the dendrimer signal detection reagent ideal for high-throughput functional genomics research. PMID- 11015605 TI - Cloning a chloride conductance mediator from the apical membrane of porcine ileal enterocytes. AB - Attempts to attribute ileal brush-border chloride conductance to specific proteins were pursued by screening a porcine intestinal cDNA library. A 0.94-kb clone was identified on expression screening with a monoclonal antibody that inhibited enterocyte brush-border chloride conductance. Further screening approaches led to the isolation of a 3.1-kb full-length sequence called pCLCA1, consistent with the identification of a 2.9-kb transcript through Northern analysis. This sequence had significant homology to the CLCA gene family of calcium-regulated chloride channels, especially to hCLCA1. However, a strong A kinase consensus phosphorylation site in a predicted cytoplasmic loop of the protein was a notable difference from the hCLCA1 gene product. Several porcine exocrine epithelial tissues, including ileum, trachea, and the major salivary glands express pCLCA1 mRNA. In situ hybridization studies localized the expression of pCLCA1 mRNA to the crypt and villus epithelia of porcine ileum, whereas tracheal expression was observed in both surface epithelium and submucosal glands. In situ expression of pCLCA1 in mouse 3T3 cells induces an ionomycin-dependent chloride conductance activity in these cells. PMID- 11015606 TI - Interaction of genetic and environmental programming of the leptin system and of obesity disposition. AB - Possible adverse interactions between an usually inconspicuous genetic trait and early environmental factors favoring the development of obesity were investigated in rats heterozygous for the leptin receptor defect "fatty" (fa). Pups were exposed to early postnatal overfeeding by reducing litter size from normally 10 12 to only 4. Rearing +/+ and +/fa pups from day 3 to 21 in small litters increased fat-free dry mass and body fat, but only in the latter did a significant interaction with genotype occur. Pronounced differences in the responsiveness of +/+ and +/fa pups to "prophylactic" leptin treatment (from day 1 to 21) were observed, with +/fa females from small litters being nearly as fat and unresponsive as previously reported for normally reared fa/fa pups. Clear heterozygous differences in total hypothalamic leptin binding, but no litter size effect, paralleling the differences in leptin responsiveness, were observed. By early postnatal overfeeding an usually inconspicuous genetic trait may thus become etiologic for the development of obesity via physiological changes other than the decreased leptin binding characterizing the genetic defect. PMID- 11015607 TI - Bradykinin B2 null mice are prone to renal dysplasia: gene-environment interactions in kidney development. AB - Congenital abnormalities of the kidney and urinary tract are a common cause of end-stage renal disease in children. Host and environment factors are implicated in the pathogenesis of aberrant renal development. However, direct evidence linking gene-environment interactions with congenital renal disease is lacking. We report an animal model of renal dysgenesis that is dependent on a defined genetic defect and specific embryonic stressor. Specifically, mice that are deficient in the bradykinin type 2 receptor gene (B(2)) and salt loaded during embryogenesis acquire an aberrant kidney phenotype and die shortly after birth. In contrast, B(2) mutant mice maintained on normal sodium intake or salt-loaded wild-type mice do not develop kidney abnormalities. The kidney abnormality is evident histologically on embryonic day 16, shortly after the onset of metanephric B(2) gene expression, and consists of distorted renal architecture, foci of tubular dysgenesis, and cyst formation. The dysplastic tubules are of distal nephron origin [Dolichos biflorus agglutinin (DBA)- and aquaporin-2 (AQP2) positive, and angiotensinogen negative]. Neonatal antihypertensive therapy fails to ameliorate the renal abnormalities, arguing against the possibility that the nephropathy is a consequence of early hypertension. Moreover, the nephropathy is intrinsic to the embryo, because B(2) homozygous offspring from heterozygous parents exhibit the same renal phenotype as offspring from homozygous null parents. Further characterization of the renal phenotype revealed an important genetic background effect since the penetrance of the congenital nephropathy is increased substantially upon backcrossing of 129/BL6 B(2) mutants to a uniform C57BL/6J. We conclude that the type 2 bradykinin receptor is required for the maintenance of metanephric structure and epithelial integrity in the presence of fetal stress. This study provides a "proof-of-principle" that defined gene environment interactions are a cause of congenital renal disease. PMID- 11015608 TI - Differential effects of dystrophin and utrophin gene transfer in immunocompetent muscular dystrophy (mdx) mice. AB - Duchenne muscular dystrophy (DMD) is a fatal disease caused by defects in the gene encoding dystrophin. Dystrophin is a cytoskeletal protein, which together with its associated protein complex, helps to protect the sarcolemma from mechanical stresses associated with muscle contraction. Gene therapy efforts aimed at supplying a normal dystrophin gene to DMD muscles could be hampered by host immune system recognition of dystrophin as a "foreign" protein. In contrast, a closely related protein called utrophin is not foreign to DMD patients and is able to compensate for dystrophin deficiency when overexpressed throughout development in transgenic mice. However, the issue of which of the two candidate molecules is superior for DMD therapy has remained an open question. In this study, dystrophin and utrophin gene transfer effects on dystrophic muscle function were directly compared in the murine (mdx) model of DMD using E1/E3 deleted adenovirus vectors containing either a dystrophin (AdV-Dys) or a utrophin (AdV-Utr) transgene. In immunologically immature neonatal animals, AdV-Dys and AdV-Utr improved tibialis anterior muscle histopathology, force-generating capacity, and the ability to resist injury caused by high-stress contractions to an equivalent degree. By contrast, only AdV-Utr was able to achieve significant improvement in force generation and the ability to resist stress-induced injury in the soleus muscle of immunocompetent mature mdx animals. In addition, in mature mdx mice, there was significantly greater transgene persistence and reduced inflammation with utrophin compared to dystrophin gene transfer. We conclude that dystrophin and utrophin are largely equivalent in their intrinsic abilities to prevent the development of muscle necrosis and weakness when expressed in neonatal mdx animals with an immature immune system. However, because immunity against dystrophin places an important limitation on the efficacy of dystrophin gene replacement in an immunocompetent mature host, the use of utrophin as an alternative to dystrophin gene transfer in this setting appears to offer a significant therapeutic advantage. PMID- 11015609 TI - Microsatellite marker panels for use in high-throughput genotyping of mouse crosses. AB - Several microsatellite genotyping panel sets have been developed that are polymorphic between C57BL/6J and CAST/Ei mice, or C57BL/6J and DBA/2J. One set of markers for each strain pair has an intermarker distance of approximately 20 cM, and a second set has an intermarker distance of 5 cM. The 20-cM set contains 105 markers for C57BL/6J x DBA/2J and 108 for C57BL/6J x CAST/Ei, divided into 13 panels. Each 5-cM set includes 350 markers arranged into 45 panels. A panel contains a number of primer pairs whose fluorescently labeled PCR products can be pooled together and separated on one lane of a polyacrylamide gel. The sets are arranged by the size of the PCR product and by the type of fluorescent dye; 5-cM sets are also arranged by chromosomal region. The 20-cM sets are most useful for full-genome scans, the 5-cM sets are useful for full-genome and/or for region specific chromosome screens. Both sets were proven as useful tools for speed congenic development, quantitative trait loci (QTL) analysis and physical mapping. These panel sets provide a throughput of 1,536-2,304 mouse genotypes daily per one gel-based system. Whole genome scans of one animal require 13 or 48 gel lanes, with 20 cM or 5 cM density, respectively. PMID- 11015610 TI - Gene expression in hypothalamus and brown adipose tissue of mice divergently selected for heat loss. AB - Gene expression was evaluated in mice divergently selected for 16 generations for heat loss, measured by direct calorimetry. The high (MH) heat loss line has approximately 50% greater heat loss, approximately 35% less body fat, approximately 20% greater feed intake, and twofold greater activity levels than the low (ML) heat loss line. At 11 wk, inbred males (developed from MH and ML) were euthanized 3 h after dark for dissection of tissues and extraction of RNA. Differential display PCR (DD-PCR) was used to evaluate transcriptional differences between lines in hypothalamus and brown adipose tissue (BAT). Evaluation was replicated within and across lines, using family pools of mRNA. Two genes were confirmed by competitive RT-PCR and/or Northern analysis to have greater levels of mRNA present in ML relative to MH mice. In both hypothalamus and BAT, the ribosomal protein L3 (RPL3) gene was expressed at higher levels in ML, whereas an unknown expressed sequence tag (EST) was also found at higher levels in the hypothalamus of ML mice. These results implicate RPL3 in regulation of energy balance and extend the genetic dissection of response to selection to the transcriptional level. PMID- 11015611 TI - Transcriptional regulation of the adipocyte fatty acid synthase gene by agouti: interaction with insulin. AB - Mice carrying dominant mutations at the agouti locus exhibit ectopic expression of agouti gene transcripts, obesity, and type II diabetes through unknown mechanisms. To gain insight into the role of agouti protein in modulating adiposity, we investigated regulation of a key lipogenic gene, fatty acid synthase (FAS) by agouti alone and in combination with insulin. Both agouti and insulin increase FAS activity in 3T3-L1 and in human adipocytes. Agouti and insulin independently and additively increase FAS activity in 3T3-L1 adipocytes. We further investigated the mechanism responsible for the agouti-induced FAS expression in these cells and demonstrated that both insulin (3-fold increase) and agouti (2-fold) increased FAS gene expression at the transcriptional level. Furthermore, insulin and agouti together exerted additive effects (5-fold increase) on FAS gene transcription. Transfection assays of FAS promoter luciferase fusion gene constructs into 3T3-L1 adipocytes indicated that the agouti response element(s) is (are) located in the -435 to -415 region (-435/ 415) of the FAS promoter. Nuclear proteins binding to this novel sequence are adipocyte specific. Thus the agouti response sequences mapped to a region upstream of the insulin-responsive element (which we previously reported to be located at -67/-52), consistent with additive effects of these two factors on FAS gene transcription. PMID- 11015612 TI - Mechanical culture conditions effect gene expression: gravity-induced changes on the space shuttle. AB - Three-dimensional suspension culture is a gravity-limited phenomenon. The balancing forces necessary to keep the aggregates in suspension increase directly with aggregate size. This leads to a self-propagating cycle of cell damage by balancing forces. Cell culture in microgravity avoids this trade-off. We determined which genes mediate three-dimensional culture of cell and tissue aggregates in the low-shear stress, low-turbulent environment of actual microgravity. Primary cultures of human renal cortical cells were flown on the space shuttle. Cells grown in microgravity and ground-based controls were grown for 6 days and fixed. RNA was extracted, and automated gene array analysis of the expression of 10, 000 genes was performed. A select group of genes were regulated in microgravity. These 1,632 genes were independent of known shear stress response element-dependent genes and heat shock proteins. Specific transcription factors underwent large changes in microgravity including the Wilms' tumor zinc finger protein, and the vitamin D receptor. A specific group of genes, under the control of defined transcription factors, mediate three-dimensional suspension culture under microgravity conditions. PMID- 11015613 TI - Large-scale analysis of gene expression changes during acute and chronic exposure to [Delta]9-THC in rats. AB - Large-scale cDNA microarrays were employed to assess transient changes in gene expression levels following acute and chronic exposure to cannabinoids in rats. A total of 24,456 cDNA clones were randomly selected from a rat brain cDNA library, amplified by PCR, and arrayed at high density to investigate differential gene expression profiles following acute (24 h), intermediate (7 days), and chronic (21 days) exposure to Delta(9)-tetrahydrocannabinol (Delta(9)-THC), the psychoactive ingredient of marijuana. Hippocampal mRNA probes labeled with (33)P obtained from both vehicle and Delta(9)-THC-treated animals were hybridized with identical cDNA microarrays. Results revealed a total of 49 different genes altered by Delta(9)-THC exposure; of these, 28 were identified, 10 had homologies to expressed sequence tags (ESTs), and 11 had no homology to known sequences in the GenBank database. Chronic or acute cannabinoid receptor activation altered expression of several genes (i.e., prostaglandin D synthase, calmodulin) involved in biochemical cascades of cannabinoid synthesis or cannabinoid effector systems. Other genes [i.e., neural cell adhesion molecule (NCAM), myelin basic protein], whose relation to cannabinoid system function was not immediately obvious, were also significantly altered. Verification of the changes obtained with the large scale screen was determined by RNA dot blots in different groups of animals treated the same as those in the large-scale screen. Results are discussed in terms of the different types of genes affected at different times during chronic Delta(9)-THC exposure. PMID- 11015614 TI - Proteoglycans in the developing brain: new conceptual insights for old proteins. AB - Proteoglycans are a heterogeneous class of proteins bearing sulfated glycosaminoglycans. Some of the proteoglycans have distinct core protein structures, and others display similarities and thus may be grouped into families such as the syndecans, the glypicans, or the hyalectans (or lecticans). Proteoglycans can be found in almost all tissues being present in the extracellular matrix, on cellular surfaces, or in intracellular granules. In recent years, brain proteoglycans have attracted growing interest due to their highly regulated spatiotemporal expression during nervous system development and maturation. There is increasing evidence that different proteoglycans act as regulators of cell migration, axonal pathfinding, synaptogenesis, and structural plasticity. This review summarizes the most recent data on structures and functions of brain proteoglycans and focuses on new physiological concepts for their potential roles in the developing central nervous system. PMID- 11015615 TI - Structure, function, and control of phosphoinositide-specific phospholipase C. AB - Phosphoinositide-specific phospholipase C (PLC) subtypes beta, gamma, and delta comprise a related group of multidomain phosphodiesterases that cleave the polar head groups from inositol lipids. Activated by all classes of cell surface receptor, these enzymes generate the ubiquitous second messengers inositol 1,4, 5 trisphosphate and diacylglycerol. The last 5 years have seen remarkable advances in our understanding of the molecular and biological facets of PLCs. New insights into their multidomain arrangement and catalytic mechanism have been gained from crystallographic studies of PLC-delta(1), while new modes of controlling PLC activity have been uncovered in cellular studies. Most notable is the realization that PLC-beta, -gamma, and -delta isoforms act in concert, each contributing to a specific aspect of the cellular response. Clues to their true biological roles were also obtained. Long assumed to function broadly in calcium-regulated processes, genetic studies in yeast, slime molds, plants, flies, and mammals point to specific and conditional roles for each PLC isoform in cell signaling and development. In this review we consider each subtype of PLC in organisms ranging from yeast to mammals and discuss their molecular regulation and biological function. PMID- 11015616 TI - Role of nitric oxide in the pathogenesis of chronic pulmonary hypertension. AB - Chronic pulmonary hypertension is a serious complication of a number of chronic lung and heart diseases. In addition to vasoconstriction, its pathogenesis includes injury to the peripheral pulmonary arteries leading to their structural remodeling. Increased pulmonary vascular synthesis of an endogenous vasodilator, nitric oxide (NO), opposes excessive increases of intravascular pressure during acute pulmonary vasoconstriction and chronic pulmonary hypertension, although evidence for reduced NO activity in pulmonary hypertension has also been presented. NO can modulate the degree of vascular injury and subsequent fibroproduction, which both underlie the development of chronic pulmonary hypertension. On one hand, NO can interrupt vascular wall injury by oxygen radicals produced in increased amounts in pulmonary hypertension. NO can also inhibit pulmonary vascular smooth muscle and fibroblast proliferative response to the injury. On the other hand, NO may combine with oxygen radicals to yield peroxynitrite and other related, highly reactive compounds. The oxidants formed in this manner may exert cytotoxic and collagenolytic effects and, therefore, promote the process of reparative vascular remodeling. The balance between the protective and adverse effects of NO is determined by the relative amounts of NO and reactive oxygen species. We speculate that this balance may be shifted toward more severe injury especially during exacerbations of chronic diseases associated with pulmonary hypertension. Targeting these adverse effects of NO-derived radicals on vascular structure represents a potential novel therapeutic approach to pulmonary hypertension in chronic lung diseases. PMID- 11015617 TI - Proximal tubular phosphate reabsorption: molecular mechanisms. AB - Renal proximal tubular reabsorption of P(i) is a key element in overall P(i) homeostasis, and it involves a secondary active P(i) transport mechanism. Among the molecularly identified sodium-phosphate (Na/P(i)) cotransport systems a brush border membrane type IIa Na-P(i) cotransporter is the key player in proximal tubular P(i) reabsorption. Physiological and pathophysiological alterations in renal P(i) reabsorption are related to altered brush-border membrane expression/content of the type IIa Na-P(i) cotransporter. Complex membrane retrieval/insertion mechanisms are involved in modulating transporter content in the brush-border membrane. In a tissue culture model (OK cells) expressing intrinsically the type IIa Na-P(i) cotransporter, the cellular cascades involved in "physiological/pathophysiological" control of P(i) reabsorption have been explored. As this cell model offers a "proximal tubular" environment, it is useful for characterization (in heterologous expression studies) of the cellular/molecular requirements for transport regulation. Finally, the oocyte expression system has permitted a thorough characterization of the transport characteristics and of structure/function relationships. Thus the cloning of the type IIa Na-P(i )cotransporter (in 1993) provided the tools to study renal brush border membrane Na-P(i) cotransport function/regulation at the cellular/molecular level as well as at the organ level and led to an understanding of cellular mechanisms involved in control of proximal tubular P(i) handling and, thus, of overall P(i) homeostasis. PMID- 11015618 TI - Dynamics and consequences of potassium shifts in skeletal muscle and heart during exercise. AB - Since it became clear that K(+) shifts with exercise are extensive and can cause more than a doubling of the extracellular [K(+)] ([K(+)](s)) as reviewed here, it has been suggested that these shifts may cause fatigue through the effect on muscle excitability and action potentials (AP). The cause of the K(+) shifts is a transient or long-lasting mismatch between outward repolarizing K(+) currents and K(+) influx carried by the Na(+)-K(+) pump. Several factors modify the effect of raised [K(+)](s) during exercise on membrane potential (E(m)) and force production. 1) Membrane conductance to K(+) is variable and controlled by various K(+) channels. Low relative K(+) conductance will reduce the contribution of [K(+)](s) to the E(m). In addition, high Cl(-) conductance may stabilize the E(m) during brief periods of large K(+) shifts. 2) The Na(+)-K(+) pump contributes with a hyperpolarizing current. 3) Cell swelling accompanies muscle contractions especially in fast-twitch muscle, although little in the heart. This will contribute considerably to the lowering of intracellular [K(+)] ([K(+)](c)) and will attenuate the exercise-induced rise of intracellular [Na(+)] ([Na(+)](c)). 4) The rise of [Na(+)](c) is sufficient to activate the Na(+)-K(+) pump to completely compensate increased K(+) release in the heart, yet not in skeletal muscle. In skeletal muscle there is strong evidence for control of pump activity not only through hormones, but through a hitherto unidentified mechanism. 5) Ionic shifts within the skeletal muscle t tubules and in the heart in extracellular clefts may markedly affect excitation-contraction coupling. 6) Age and state of training together with nutritional state modify muscle K(+) content and the abundance of Na(+)-K(+) pumps. We conclude that despite modifying factors coming into play during muscle activity, the K(+) shifts with high-intensity exercise may contribute substantially to fatigue in skeletal muscle, whereas in the heart, except during ischemia, the K(+) balance is controlled much more effectively. PMID- 11015619 TI - Calcineurin: form and function. AB - Calcineurin is a eukaryotic Ca(2+)- and calmodulin-dependent serine/threonine protein phosphatase. It is a heterodimeric protein consisting of a catalytic subunit calcineurin A, which contains an active site dinuclear metal center, and a tightly associated, myristoylated, Ca(2+)-binding subunit, calcineurin B. The primary sequence of both subunits and heterodimeric quaternary structure is highly conserved from yeast to mammals. As a serine/threonine protein phosphatase, calcineurin participates in a number of cellular processes and Ca(2+)-dependent signal transduction pathways. Calcineurin is potently inhibited by immunosuppressant drugs, cyclosporin A and FK506, in the presence of their respective cytoplasmic immunophilin proteins, cyclophilin and FK506-binding protein. Many studies have used these immunosuppressant drugs and/or modern genetic techniques to disrupt calcineurin in model organisms such as yeast, filamentous fungi, plants, vertebrates, and mammals to explore its biological function. Recent advances regarding calcineurin structure include the determination of its three-dimensional structure. In addition, biochemical and spectroscopic studies are beginning to unravel aspects of the mechanism of phosphate ester hydrolysis including the importance of the dinuclear metal ion cofactor and metal ion redox chemistry, studies which may lead to new calcineurin inhibitors. This review provides a comprehensive examination of the biological roles of calcineurin and reviews aspects related to its structure and catalytic mechanism. PMID- 11015620 TI - Prolactin: structure, function, and regulation of secretion. AB - Prolactin is a protein hormone of the anterior pituitary gland that was originally named for its ability to promote lactation in response to the suckling stimulus of hungry young mammals. We now know that prolactin is not as simple as originally described. Indeed, chemically, prolactin appears in a multiplicity of posttranslational forms ranging from size variants to chemical modifications such as phosphorylation or glycosylation. It is not only synthesized in the pituitary gland, as originally described, but also within the central nervous system, the immune system, the uterus and its associated tissues of conception, and even the mammary gland itself. Moreover, its biological actions are not limited solely to reproduction because it has been shown to control a variety of behaviors and even play a role in homeostasis. Prolactin-releasing stimuli not only include the nursing stimulus, but light, audition, olfaction, and stress can serve a stimulatory role. Finally, although it is well known that dopamine of hypothalamic origin provides inhibitory control over the secretion of prolactin, other factors within the brain, pituitary gland, and peripheral organs have been shown to inhibit or stimulate prolactin secretion as well. It is the purpose of this review to provide a comprehensive survey of our current understanding of prolactin's function and its regulation and to expose some of the controversies still existing. PMID- 11015621 TI - Development of intestinal transport function in mammals. AB - Considerable progress has been made over the last decade in the understanding of mechanisms responsible for the ontogenetic changes of mammalian intestine. This review presents the current knowledge about the development of intestinal transport function in the context of intestinal mucosa ontogeny. The review predominantly focuses on signals that trigger and/or modulate the developmental changes of intestinal transport. After an overview of the proliferation and differentiation of intestinal mucosa, data about the bidirectional traffic (absorption and secretion) across the developing intestinal epithelium are presented. The largest part of the review is devoted to the description of developmental patterns concerning the absorption of nutrients, ions, water, vitamins, trace elements, and milk-borne biologically active substances. Furthermore, the review examines the development of intestinal secretion that has a variety of functions including maintenance of the fluidity of the intestinal content, lubrication of mucosal surface, and mucosal protection. The age dependent shifts of absorption and secretion are the subject of integrated regulatory mechanisms, and hence, the input of hormonal, nervous, immune, and dietary signals is reviewed. Finally, the utilization of energy for transport processes in the developing intestine is highlighted, and the interactions between various sources of energy are discussed. The review ends with suggestions concerning possible directions of future research. PMID- 11015622 TI - Role of platelet-activating factor in cardiovascular pathophysiology. AB - Platelet-activating factor (PAF) is a phospholipid mediator that belongs to a family of biologically active, structurally related alkyl phosphoglycerides. PAF acts via a specific receptor that is coupled with a G protein, which activates a phosphatidylinositol-specific phospholipase C. In this review we focus on the aspects that are more relevant for the cell biology of the cardiovascular system. The in vitro studies provided evidence for a role of PAF both as intercellular and intracellular messenger involved in cell-to-cell communication. In the cardiovascular system, PAF may have a role in embryogenesis because it stimulates endothelial cell migration and angiogenesis and may affect cardiac function because it exhibits mechanical and electrophysiological actions on cardiomyocytes. Moreover, PAF may contribute to modulation of blood pressure mainly by affecting the renal vascular circulation. In pathological conditions, PAF has been involved in the hypotension and cardiac dysfunctions occurring in various cardiovascular stress situations such as cardiac anaphylaxis and hemorrhagic, traumatic, and septic shock syndromes. In addition, experimental studies indicate that PAF has a critical role in the development of myocardial ischemia-reperfusion injury. Indeed, PAF cooperates in the recruitment of leukocytes in inflamed tissue by promoting adhesion to the endothelium and extravascular transmigration of leukocytes. The finding that human heart can produce PAF, expresses PAF receptor, and is sensitive to the negative inotropic action of PAF suggests that this mediator may have a role also in human cardiovascular pathophysiology. PMID- 11015623 TI - Emotional state, coping styles, and somatic variables in patients with chronic hepatitis C. AB - The authors in a cross-sectional study examined 113 patients with chronic hepatitis C (CHC) without widely progressed or decompensated liver disease. The patients were investigated for emotional state (depression, anxiety, coping styles) and somatic/sociodemographic variables. A high percentage of patients had positive scores for depression (22.4%) and anxiety (15.2%). Mode of acquisition (e.g., former drug abuse) and histological grade of liver damage had no significant influence on emotional state or coping strategies. Older patients (> or = 50 years) were significantly more depressed (P = 0.024). Patients with a recently diagnosed CHC (> 4 weeks, < 6 months) had significantly lower scores for depression (P = 0.003) and anxiety (P = 0.001) than the subgroup with a time interval since initial diagnosis of more than 5 years. Recently diagnosed CHC patients also showed the highest levels of problem-solving behavior. Patients who were advised not to undergo an interferon therapy were significantly more depressed (P = 0.001) and anxious (P = 0.028). Older patients with CHC and patients with a long period since CHC diagnosis or who were advised not to undergo interferon therapy should be carefully and regularly assessed for depression, anxiety, and inappropriate coping styles. PMID- 11015624 TI - An observer scale to measure alexithymia. AB - The authors developed a relatively brief observer alexithymia measure that can be used by patients' acquaintances and relatives. Items corresponding to the defining features of alexithymia (California Q-Set Alexithymia Prototype) were written, and the new instrument's psychometric properties were evaluated in 3 lay rater samples. The 33-item Observer Alexithymia Scale (OAS) is internally consistent (coefficient alphas = 0.88 and 0.89) and stable (2-week test-retest reliability = 0.87). Moreover, it has an interpretable five-factor structure (based on exploratory and confirmatory factor analyses): distant, uninsightful, somatizing, humorless, and rigid. The OAS is a reliable instrument with a stable factor structure and good conceptual coverage and thus, it appears to be a useful tool for collecting observer data on the clinically relevant expressions of alexithymia that receive minimal attention. PMID- 11015625 TI - The Structured Interview for Renal Transplantation--SIRT. AB - Despite the fact that the demand for psychiatric evaluations of organ transplant recipients is increasing, there is not a commonly agreed upon protocol that can guide clinicians. A standard psychiatric interview, although necessary, is not sufficient when interviewing transplant candidates. In addition, it is important to acquire information specific to the medical regimen associated with renal disease and renal transplantation. The purpose of this paper is to present the Structured Interview for Renal Transplantation (SIRT). The SIRT was developed as a tool to guide clinicians through the interview process by providing a comprehensive structure while still allowing for flexibility. There are many advantages of using the SIRT--it is a tool that facilitates a clinician's ability to conduct a thorough evaluation in a time-efficient manner. It is also an excellent instrument for training clinicians, and the structure of the interview is appropriate for use in research. PMID- 11015626 TI - Religious altruism and organ donation. AB - Clinicians performing psychiatric assessments of potential organ donors must consider the motivations behind an act that is--strictly in terms of its physiological implications--entirely altruistic. The authors present two case reports in which proposed kidney donors conceptualized their offers exclusively in terms of their religious beliefs and not in terms of kinship or emotional intimacy with the intended recipients. The negative reactions of some clinicians to the offers reveal the readiness with which religious beliefs can be pathologized and the way in which biological relationships can unduly restrict the clinical understanding of healthy altruism. PMID- 11015628 TI - Dimensions of psychopathology in the medically ill. A latent trait analysis. AB - The authors examined the latent structure of psychiatric symptoms occurring in patients with medical illness. Symptom data were collected from 312 hospitalized medically ill patients using the Monash Interview for Liaison Psychiatry and subjected to latent trait analysis. A model with 5 dimensions provided an acceptable fit to the data. Dimensions were characterized as demoralization, anhedonia, autonomic anxiety, somatic symptoms, and grief. The demoralization dimension was similar to the concept of demoralization described by Frank and to the "giving up-given up complex" described by Engel. The concepts of demoralization, grief, and anhedonia may be useful in increasing understanding of the minor depressions in people with medical illness and in increasing the specificity of psychological and pharmacological treatments for these disorders. PMID- 11015627 TI - Psychiatric training in primary care medicine residency programs. A national survey. AB - The authors conducted a national survey to investigate the current status of psychiatric training in primary care/internal medicine residencies. Fifty-four residency training directors completed and returned the survey. The survey results show that an average of 99 hours (69.5 hours clinical plus 29.8 hours didactics) is devoted to psychiatric training during the 3 years of primary care/internal medicine residency training. Responding residency training directors indicated that psychiatric training is important (an average of 7 out of 10 on a 10-point rating scale), and 63% of respondents indicated that more training in psychiatry is needed. PMID- 11015630 TI - The use of a transitional object in the context of medical illness. PMID- 11015629 TI - Association between depressive symptoms and mortality in medical inpatients. AB - The authors interviewed a consecutive series of medical inpatients (N = 241) using the Schedule for Affective Disorders and Schizophrenia to determine which depressive symptoms are associated with in-hospital mortality. Fifteen depressive symptoms, pain, and physical discomfort were assessed along with medical comorbidity. Twenty patients died in-hospital (8.3%). Logistic regression showed that anhedonia, hopelessness, worthlessness, indecisiveness, and insomnia predicted in-hospital death after adjusting for physical comorbidity and age. Clinicians should be aware that these depressive symptoms may predict mortality in medical inpatients. Future studies should address which treatment modalities lead to better outcomes. PMID- 11015631 TI - Seizure disorder is in the differential diagnosis of panic disorder. PMID- 11015632 TI - Prophylactic treatment of depression induced by interferon-alpha. PMID- 11015633 TI - Breast cancer, bipolar disorder, catatonia, and life-preserving electroconvulsive therapy. PMID- 11015634 TI - Steroid-induced mania in poststroke patient involving the right basal ganglion and right frontal region. PMID- 11015635 TI - Delusional parasitosis accompanied by word deafness due to cerebral infarction: folie a deux. PMID- 11015636 TI - Severe and moderate hypertriglyceridemia secondary to citalopram and fluoxetine. PMID- 11015637 TI - Diagnosing demoralization in consultation psychiatry. PMID- 11015638 TI - Illusion or hallucination? Cholecystitis presenting as pseudopregnancy in schizophrenia. PMID- 11015639 TI - Hepatotoxicity with combination of valproic acid, ritonavir, and nevirapine: a case report. PMID- 11015641 TI - Intravenous valproate treatment of severe manic symptoms after gastric bypass surgery: a case report. PMID- 11015640 TI - Neuroleptic malignant syndrome associated with use of risperidone, ritonavir, and indinavir: a case report. PMID- 11015642 TI - [XVIII Congress of the French Society of Endocrinology. 4-7 October 2000. Brest, France. Abstracts]. PMID- 11015643 TI - [On the guidelines concerning correct anatomic pathology and cytopathology practice by the Union of Belgian Pathologists of Medical Specialties and Pathologic Anatomy]. PMID- 11015644 TI - [Emergence of new infectious disease: the microbiologist's point of view]. PMID- 11015645 TI - [Emergence of new infectious disease: the anatomic pathologists's point of view]. PMID- 11015646 TI - [Clear cell soft tissue sarcoma. Clinical, histopathological and prognostic study of 36 cases]. AB - Thirty six cases of clear cell sarcoma of soft tissue are reported. The median age was 44 years (5 to 80 years). The principal sites were the foot (11 cases), the hand and wrist (7 cases) and the knee (6 cases). The architecture was fascicular with lobular arrangement of cells delimited by delicate fibrous septa intimately bound to tendons or aponeuroses. Tumoral cells were round or fusiform with abundant clear cytoplasm sometimes epithelioid with round nuclei and prominent nucleoli. The mitotic rate was evaluated to 9/10 HPF. S100 protein was expressed in 33/36 cases and HMB45 marked 29/31 cases, without expression of cytokeratin. Three-year and 5-year survival rate were respectively 72% and 62%. Prognosis factors for global survival were efficiency of initial treatment with distal location and necrosis and FNCLCC grade. The distinction of clear cell sarcoma from metastatic melanoma is important because of the difference of prognosis. PMID- 11015647 TI - [Evaluation of the effect of pathology slide review on clinical practice in an oncology center]. AB - Our aim was to evaluate the clinical impact of pathology review in an oncology center, in which review is not performed for every patient. This retrospective study involved 100 consecutive patients, whose slides were reviewed in our center. A standardized data sheet was filled out by oncologists for each patient. Pathology review was considered as responsible for a major (35%), moderate (40%), or mild or no (25%) modification of clinical practice. Modification concerned either initial investigations, treatment or medical follow up, and was independent of the reason for which review was performed. Eleven patients underwent a second biopsy. Whatever the possible discrepancies between initial and review diagnosis, our results show that pathological review has a major influence on clinical practice in patients with cancer. PMID- 11015648 TI - [Lichen striatus: evaluation of histologic criteria in 13 cases]. AB - Lichen striatus (LS) is an uncommon dermatosis that generally affects children. Sometimes biopsy is required to assess the diagnosis. It is usually mentioned that LS has no specific histopathological criteria. Recently Gianotti et al., in 50% of the examined cases, found clues to the diagnosis. We have evaluated 13 biopsies of children affected by LS, in order to evaluate histopathological criteria to allow a diagnosis. The features nearly constantly present (12/13) in each specimen were focally band-like lymphocytic infiltrate with variable exocytosis and necrotic keratinocytes within the epidermis surrounded by lymphocytes. Epidermis hyperplasia, sometimes psoriasiform, was also present. In addition, one of remarkable the features of LS was the alignment of the infiltrate along eccrine ducts and glands and/or follicles. In 3 cases the deep lymphocytic infiltration was particularly dense. These results confirm that helpful clues to histopathological diagnosis of LS exist. PMID- 11015649 TI - [Roles and mechanisms of apoptosis in infectious diseases]. PMID- 11015650 TI - [Emerging infectious diseases]. PMID- 11015651 TI - [Disseminated mucormycosis in AIDS]. AB - Mucormycosis is a rare often fatal opportunistic condition. It mainly occurs in immunocompromised subjects but rarely in AIDS patients. An eighteen-year-old HIV+ man from Zaire died rapidly from disseminated mucormycosis with pulmonary, cardiac, renal, hepatic, splenic and gastric involvement. Fungi were observed in all these organs with a particular vascular trophism. Rhizopus orizae was identified on cultures. Rapid microscopic diagnosis, in the localized stage, could improve the poor prognosis of this infection. PMID- 11015652 TI - [Cutaneous nodular infection with hemopathy]. AB - A 36-year-old man with treated hairy cell leukemia developed a skin infection due to Mycobacterium marinum. A spectrum of atypical cutaneous mycobacteriosis presentations with immunosuppression can be found. The recognition of the disease needs cutaneous biopsies for histopathology and its identification by specific laboratory methods to adapt treatment. PMID- 11015653 TI - [Endolymphatic sac tumor: a rare tumor of internal ear. Report of two cases]. AB - Papillary tumors of the temporal bone are rare and aggressive neoplasms. Recently described, these tumors had initially a presumed middle-ear origin. Only recently, convincing anatomic, morphological and immunohistochemical arguments exist for an endolymphatic sac origin (inner-ear origin). We report two cases of endolymphatic sac tumor. These tumors can be encountered sporadically or in Von Hippel-Lindau disease. They classically grow very slowly, resulting in late clinical manifestations with expansive mass invading temporal bone and extending in posterior fossa. Radiologically, these endolymphatic sac tumors can mimic metastatic carcinoma, paraganglioma, or cerebellar haemangioblastoma specially in von Hippel-Lindau disease. Histology shows a papillary epithelial tumor with hypervascular stroma, without atypia. The treatment for these tumors is surgical and curative when early diagnosed. In apparently sporadic cases, genetic analysis for Von Hippel-Lindau disease should be considered. PMID- 11015654 TI - [A case of primitive meningeal rhabdomyosarcoma. Histological, immunohistochemical and ultrastructural study]. AB - A 64 year-old patient, complained of headache and neurological disorders. CT scan found a voluminous solitary tumor of the posterior part of the left cavernous sinus. Removal of tumor was followed by a rapid recurrence and by the patient's death. Histologic study found a malignant undifferentiated tumoral proliferation, with strap-like cells. Immunohistochemical stains were positive for conjunctival and muscular differentiation. Ultrastructural study revealed intracytoplasmic filamentous striated structure. The primary meningeal rhabdomyosarcoma is an exceptional tumor, generally affecting young patients. Its prognosis is poor and its histogenesis remains unclear. PMID- 11015655 TI - [Primary large cell neuroendocrine carcinoma of the kidney: morphologic and immunohistochemical features of two cases]. AB - Primary neuroendocrine carcinomas of the kidney are infrequent. The large cell neuroendocrine carcinoma is a subtype with an aggressive course and a worse prognosis. We report two cases, a 35-year-old man and a 75-year-old woman who died within 6 and 5-months after surgery despite radical nephrectomy and chemotherapy. Histological and immunohistochemical features are presented. PMID- 11015656 TI - [Intraluminal renal metastasis from a rectal adenocarcinoma: an unusual site]. AB - Primary adenocarcinoma of the urinary tract are uncommon. But secondary involvement of pyelocalyceal system by metastasis of colorectal origin is rare. We report a case of late rectal metastasis with renal pelvis growth presenting as a pyonephrosis. This study emphasizes the relevance of cytokeratin 7 and 20 immunostaining in such differential diagnosis. PMID- 11015657 TI - [Adenomyoepithelioma of the breast: two cases]. AB - Adenomyoepithelioma of the breast are very rare and form a heterogeneous tumoral group in terms of their morphology and biological behaviour. We report two adenomyoepitheliomas of the breast. One appeared in a 29-year-old woman and was a tubular adenomyoepithelioma. The patient was free of disease at a 22 months follow-up. The other was a lobular adenomyoepithelioma that appeared in a 76-year old woman and was characterized by its high mitotic rate and its association with myoepitheliosis. Adenomyoepitheliomas of the breast are currently classified as low grade malignant tumors that may recur and rarely metastasize, but their histoprognostic factors are not yet defined. PMID- 11015658 TI - [Adenomatoid tumor of the pleura. Case report]. AB - We report a case of an adenomatoid tumor of particular location within the pleura, incidentally discovered on a pulmonary lobectomy specimen after surgical resection of a pulmonary squamous cell carcinoma. This adenomatoid tumor appeared as a unique pleural mass located away from the primary carcinoma and consisted of a cellular proliferation organised in tubes and sheets. Adenomatoid tumors are considered as benign tumors of mesothelial nature. Their morphological and immunohistochemical features in association with their location to the pleura, warrant a precise analysis to eliminate malignant tumours such as malignant mesothelioma or metastatic adenocarcinoma. PMID- 11015659 TI - [A case study of Woringer-Kolopp disease. Immunohistochemical study]. AB - Woringer-Kolopp disease is a localized epidermotropic T cell lymphoma with good prognosis. We describe a 79-year-old man with an erythematous scaly plaque of the foot. Clinical diagnosis of psoriasis, parapsoriasis and fungal infection were proposed. Histopathological and immunohistochemical findings were characteristic of Woringer-Kolopp disease. We observed an immunohistochemical positivity of tumour cells for the anti-CD103 antibody (alphaEB7 integrin) according to the epidermotropism of this localized cutaneous T cell lymphoma. PMID- 11015660 TI - [Tender nodule of the right hand. What is your diagnosis? Diagnosis: Intra vascular nodular fasciitis]. PMID- 11015661 TI - [A tumoral pitfall. What is your diagnosis? Diagnosis: Chordoid meningioma]. PMID- 11015662 TI - [An uncommon mastoid cyst. What is your diagnosis? Diagnosis: ceruminous cystic adenoma]. PMID- 11015663 TI - [An ulcerative tumor of the fundic mucosa. What is your diagnosis? Diagnosis: Gastric plasma cell granuloma]. PMID- 11015664 TI - [An acid rectal polyp. What is your diagnosis? Diagnosis: Gastric heterotopia of the rectum]. PMID- 11015665 TI - [A standardised histopathological worksheet for the histopathologic examination of primary cutaneous melanomas. Experiences at the Gustave-Roussy Institute]. AB - A standardised worksheet for the histopathology report of primary cutaneous melanoma used at the Gustave-Roussy Institute is described. Each item is discussed. PMID- 11015666 TI - [A proposal for a method of reporting cervico-uterine smears]. AB - A method for reporting Papsmears is proposed. Paperwork is limited to choosing a few codes on a working list prepared according to the Bethesda system and the ADICAP coding system. The method is easy to use. It reduces the work load of time for cytologists and secretaries. It allows harmonization and structurization of the look and the filling of reports for Papsmears. It may be easily adapted for vaginal smears. It leads to a report complete and adapted to the Bethesda system. It gives the possibility of adding comments and additional codes. It avoids most mistakes about obligatory codes. It makes easier to perform statistical evaluation and to initiate a quality control program. PMID- 11015667 TI - [Recent advance in the classification of tumors using microarrays]. PMID- 11015668 TI - [Metachronous bilateral germ cell tumors of the testis]. PMID- 11015669 TI - [A diagnostic pitfall in the pathology of bladder tumors: the nested variant of urothelial carcinoma]. PMID- 11015670 TI - [A misleading mistake in the classification of colonic and rectal cancers, in the 4th edition of the TNM Atlas, published in French by Springer]. PMID- 11015671 TI - [Neurofibromatosis type 2. Preliminary results of gamma knife radiosurgery of vestibular schwannomas]. AB - BACKGROUND AND PURPOSE: The aim of this study was to assess tolerance and efficacy of gamma knife radiosurgery on vestibular schwannomas for patients affected with neurofibromatosis type 2. METHODS: Between July 1992 and December 1997, a gamma knife procedure was performed on 35 vestibular schwannomas affecting 27 patients (12 females and 15 males, mean age=27 years-old, range: 14 65). Fifteen of the patients were included in the Wishart subtype (severe form) and 12 patients in the Gardner subtype (mild form). This group of 27 patients represented 8,2% of the total group of vestibular schwannomas radiosurgically treated by our team. The mean tumor volume was 4,000 mm(3) (range: 400-14,400 mm(3)) and staging according to Koos classification was 9 stage 2 tumors (extension in the cerebellopontine angle), 19 stage 3 tumors (in contact with the brain stem or cerebellum) and 7 stage 4 tumors (compression of axial structures). The delivered mean marginal dose (50% isodose) was 13 Gy (range: 10-18 Gy). After the treatment, the mean clinical and radiological follow-up was 32 months (range: 6-70). RESULTS: Twenty six (74%) of the treated tumors were controlled by the treatment (15 stabilizations and 11 regressions of the tumor volume) at last follow-up. One microsurgical removal was required in a growing stage 4 tumor and in 2 cases of growing stage 3 tumors. Three post-radiosurgical facial nerve deficits (9%) were observed, 2 of them were transient. According to the Gardner and Robertson classification, classes I (good) and II (serviceable) hearing were preserved at last follow-up in 57% of the patients having the same hearing level prior to the gamma knife. CONCLUSIONS: Our experience confirms that tolerance of gamma knife radiosurgery compares favorably with microsurgery of bilateral vestibular schwannomas. This treatment should be restricted to small and medium growing tumors. Treatment strategy of neurofibromatosis type 2 patients should be planned by multidisciplinary experienced teams disposing of the whole armamentarium. A longer follow-up study is required to confirm the current results regarding the tumor control rate. PMID- 11015672 TI - The carpal tunnel syndrome. Anatomo-clinical correlations. AB - We review a series of 1,280 operated cases of carpal tunnel syndrome (CTS), with an unexpected high frequency of transverse muscular fibers within the carpal canal (11%), particularly in a group of young male patients under 30 years of age. This abnormal presence of muscle fibers could explain the early development of CTS in young male workers. PMID- 11015673 TI - [Hearing preservation in vestibular schwannoma surgery: indications, techniques and results in the literature since 1990]. AB - Many authors have recently reported hearing preservation for approximately two thirds of acoustic neuroma patients. The results have driven them to recommend early surgery for all patients whose hearing might be saved. Inversely, other authors advocate that MRI-screening may be beneficial in some patients and suggest surgery only in case of tumor growth or progressive hearing loss. In order to facilitate therapeutic decision making, we reviewed the techniques and results in the main papers dealing with hearing preservation in the course of acoustic neuroma surgery published since 1990. This analysis showed that the average percentage of useful hearing preservation is about 31%. It is however difficult to compare the different series due to the different methodologies used. This emphasizes the need for standardization of hearing selection and reporting criteria to a) identify factors predictive of success (tumor characteristics, preoperative hearing, monitoring, surgical pathways), and b) elaborate well accepted decisional guidelines (early surgery or MRI screening), notably for small and non-symptomatic tumors which show increasing prevalence. PMID- 11015674 TI - [Apoptosis and pathologies of the central nervous system]. AB - The program of cell death called apoptosis is a biochemical and genetical pathway which has been well conserved throughout evolution. In the past years, the involvement of apoptosis has been shown not only during the embryonic development of the nervous system, but also in neurodegenerative diseases. Moreover, resistance to apoptosis is clearly an important factor in tumor growth and seems to be, at least partially, a major process in chemo- and radio-resistance. In this review, we briefly describe the main molecular events of the apoptotic process, known to date, and discuss possible defects in the central nervous system diseases. PMID- 11015675 TI - [Arachnoid cysts of the middle fossa and associated subdural hematoma. Three case reports and review of the literature]. AB - BACKGROUND AND PURPOSE: We report 3 cases of subdural hematoma associated with arachnoid cyst of the middle fossa, and discuss the treatment. METHODS: Three males aged 33, 63 and 68 were treated in our institution. Before this event the cyst was asymptomatic and unknown. Magnetic resonance imaging was the most contributive radiologic exam. Surgical procedure was limited to subdural hematoma evacuation. The internal wall of subdural hematoma was opened in all cases. RESULTS: The clinical outcome was good for all patients. The CT or MRI scan follow-up 3 months later revealed nearly total disappearance of subdural hematoma for 2 out of 3 cases. For one patient the volume of arachnoid cyst decreased. CONCLUSIONS: We propose subdural hematoma drainage without any specific treatment (shunt or fenestration) of the arachnoid cyst, for this category of patient. PMID- 11015676 TI - [Spontaneous temporo-sphenoidal encephalocele. A case report]. AB - A case of anteromedial temporo-sphenoidal meningo-encephalocele in a 43-year-old woman is reported. She was referred to us after a five-year history of rhinorrhea of CSF. Three years ago, she presented an epileptic seizure. CT scan showed an empty sella and an opacity of the right part of the sphenoid sinus. On coronal sections, a defect was visible in the superior wall of the right lateral recess of the sphenoid sinus. The lateral sphenoidal rhinorrhea was confirmed by a CT cisternography. The patient was operated on via a frontotemporal approach with an orbitozygomatic removal. The dural defect was filled by a patch of pericranium. The patient is without evidence of cerebrospinal fluid leak 2.5 years later. Fifteen cases of anteromedial temporosphenoidal meningo-encephaloceles have been reported. They probably are of a developmental origin. Some cases have been successfully treated via a transsphenoidal route. However, surgical failures related to this approach can be due to the lack of control of the lateral recess of the sphenoid sinus. Therefore, the frontotemporal approach seems more adequate. PMID- 11015677 TI - [Epidural hydatid cyst of the posterior fossa]. AB - We report a case of epidural hydatid cyst in the posterior fossa in a 5-year-old child. The disease was revealed by raised intracranial pressure with torticollis. The diagnosis was based on the brain CT scan and MRI, and confirmed surgically. The course was uneventful. Cranial epidural hydatid cysts are very rare: only 18 cases have been reported previously in the literature. PMID- 11015678 TI - [Synovial cyst of cervical spine]. AB - The synovial cyst is a rare cause of cervical medullar compression. We report on one case of a 74-year-old man hospitalized for electric discharge in the limbs at neck extension. The MRI and CT-scan revealed a posterior extradural cystic lesion at the level of C5-C6. The surgical excision of the lesion resulted in completed recovery. The histological examination confirmed the diagnosis of a synovial cyst. Through this case and the review of the 9 reported cases in the literature, we discuss the etiopathogenesis, diagnosis and treatment of the lower cervical synovial cyst. PMID- 11015679 TI - [Cerebellar hemorrhage complicating a supratentorial craniotomy. A case report and review of the literature]. AB - Postoperative cerebellar hemorrhage after a supratentorial craniotomy represents a rare event. We report a case of a patient with a meningioma of the jugum who developed suddenly after surgery a neurological deterioration due to a cerebellar hemorrhage detected on the CT scan. An occipital craniectomy and an external ventricular drainage were performed in emergency. A complete neurological recovery was observed after surgery. Nineteen similar other cases are found in the literature. Pre- and postoperative high blood pressure, lowered intracranial pressure and mispositioning of the head during surgery could be at the origin of the hemorrhage. Size of the hemorrhage, time between diagnosis and treatment represent two prognostic factors. All patients, who present a neurological deterioration in postoperative course, must have CT scan including posterior fossa. PMID- 11015680 TI - [Aneurysm of the postero-inferior cerebellar artery associated with arteriovenous malformation]. AB - A saccular aneurysm of the posterior inferior cerebellar artery lying in the fourth ventricle and an arteriovenous malformation in the cerebellum fed by the same artery are reported. Clipping of the neck of the aneurysm and total removal of the arteriovenous malformation were performed successfully during one stage. Review of the pertinent literature indicates that the aneurysm of the posterior inferior cerebellar artery originates from the peripheral portion of the artery when associated with arteriovenous malformation. The treatment is often surgical with good results in more than 80% of cases. A one-stage operation is the method of choice. Embolization is not without risk and can be attempted when the PICA's course allows easy catheterization. PMID- 11015681 TI - [Unique intracerebral plasmacytoma: a case possibly induced by head trauma]. PMID- 11015682 TI - [Guidelines for antibiotic prophylaxis in surgery patients: application to Neurosurgery. 1999 update. Working Group of Experts of the Societe Francaise d'Anesthesie et de Reanimation (SFAR)]. PMID- 11015683 TI - Transporter-mediated permeation of drugs across the blood-brain barrier. AB - Drug distribution into the brain is strictly regulated by the presence of the blood-brain barrier (BBB) that is formed by brain capillary endothelial cells. Since the endothelial cells are connected to each other by tight junctions and lack pores and/or fenestrations, compounds must cross the membranes of the cells to enter the brain from the bloodstream. Therefore, hydrophilic compounds cannot cross the barrier in the absence of specific mechanisms such as membrane transporters or endocytosis. So, for efficient supply of hydrophilic nutrients, the BBB is equipped with membrane transport systems and some of those transporter proteins have been shown to accept drug molecules and transport them into brain. In the present review, we describe mainly the transporters that are involved in drug transfer across the BBB and have been molecularly identified. The transport systems described include transporters for amino acids, monocarboxylic acids, organic cations, hexoses, nucleosides, and peptides. Most of these transporters function in the direction of influx from blood to brain; the presence of efflux transporters from brain to blood has also been demonstrated, including P glycoprotein, MRPs, and other unknown transporters. These efflux transporters seem to be functional for detoxication and/or prevention of nonessential compounds from entering the brain. Various drugs are transported out of the brain via such efflux transporters, resulting in the decrease of CNS side effects for drugs that have pharmacological targets in peripheral tissues or in the reduction of efficacy in CNS because of the lower delivery by efflux transport. To identify the transporters functional at the BBB and to examine the possible involvement of them in drug transports by molecular and physiological approaches will provide a rational basis for controlling drug distribution to the brain. PMID- 11015684 TI - Statistical properties of thermodynamic quantities for cyclodextrin complex formation. AB - Literature values of DeltaG degrees (change in Gibbs free energy), DeltaH degrees (change in enthalpy), and TDeltaS degrees (temperature times change in entropy) for 1:1 complex formation by alpha-, beta-, and gamma-cyclodextrins constitute normally distributed populations with the following statistical parameters (all energy quantities in kcal mol(-1); n is the number of data points; mu is the population mean; sigma is the standard deviation): for alpha-cyclodextrin, n = 512, micro(DeltaG) = -2.85, sigma(DeltaG) = 1.23, micro(DeltaH) = -4.77, sigma(DeltaH) = 2.98, micro(TDeltaS) = -1.96, and sigma(TDeltaS) = 2.72; for beta cyclodextrin, n = 415, micro(DeltaG) = -3.67, sigma(DeltaG) = 1. 37, micro(DeltaH) = -4.24, sigma(DeltaH) = 2.89, micro(DeltaS) = -0. 56, and sigma(TDeltaS) = 2.63; for gamma-cyclodextrin, n = 42, micro(DeltaG) = -3.71, sigma(DeltaG) = 1.19, micro(DeltaH) = -3.10, sigma(DeltaH) = 3.39, micro(TDeltaS) = +0.69, and sigma(TDeltaS) = 3. 29. The temperature is 298.15 K. The mean DeltaG degrees values correspond to binding constants of 123, 490, and 525 M(-1) for alpha-, beta-, and gamma-cyclodextrins, respectively. PMID- 11015685 TI - Intrinsic absolute bioavailability prediction in rats based on in situ absorption rate constants and/or in vitro partition coefficients: 6-fluoroquinolones. AB - A preliminary study attempting to predict the intrinsic absolute bioavailability of a group of antibacterial 6-fluoroquinolones-including true and imperfect homologues as well as heterologues-was carried out. The intrinsic absolute bioavailability of the test compounds, F, was assessed on permanently cannulated conscious rats by comparing the trapezoidal normalized areas under the plasma concentration-time curves obtained by intravenous and oral routes (n = 8-12). The high-performance liquid chromatography analytical methods used for plasma samples are described. Prediction of the absolute bioavailability of the compounds was based on their intrinsic rat gut in situ absorption rate constant, k(a). The working equation was: where T represents the mean absorbing time. A T value of 0.93 (+/-0.06) h provides the best correlation between predicted and experimentally obtained bioavailabilities (F' and F, respectively) when k(a) values are used (slope a = 1.10; intercept b = -0.05; r = 0.991). The k(a) values can also be expressed in function of the in vitro partition coefficients, P, between n-octanol and a phosphate buffer. In this case, theoretical k(a) values can be determined with the parameters of a standard k(a)/P correlation previously established for a group of model compounds. When P values are taken instead of k(a) values, reliable bioavailability predictions can also be made. These and other relevant features of the method are discussed. PMID- 11015686 TI - Impact of mechanism-based enzyme inactivation on inhibitor potency: implications for rational drug discovery. AB - Mechanism-based enzyme inactivators (MBEIs) have unique kinetic actions that make predictions of potency, selectivity, and potential for metabolic drug interactions more complex than for competitive antagonists. We have derived a mathematical relationship that links the influence of substrate concentration and binding constant ([S] and K(m), respectively), inhibitor concentration and binding constant ([I] and K(I), respectively), and inactivation rate constant (k(inact)) to enzyme activity (v) and maximal activity (V(max)) at any time (t). The kinetic behavior of this relationship was validated in murine-macrophage cell cultures using MBEIs of nitric oxide synthase (NOS). This initial equation was also used in the derivation of a new relationship that directly links the kinetic parameters of mechanism-based inactivation to inhibitory potency at a particular time (IC((t))(50)). Using this direct relationship, we observed that the predicted rank inhibitory potency of a series of MBEIs was improved over that predicted by the K(I) parameter alone. These relationships offer a fundamental understanding of the kinetics of MBEI action and may be useful in the evaluation of these compounds during the discovery process. PMID- 11015687 TI - Prediction of transdermal flux of prodrugs of 5-fluorouracil, theophylline, and 6 mercaptopurine with a series/parallel model. AB - Multiple regression analysis of fluxes from suspensions in isopropyl myristate (J(M)) as a function of molecular weights (MW) and solubilities in isopropyl myristate (S(IPM)) and water (S(AQ)) were performed on a data set of 41 compounds (n = 41) comprising 39 prodrugs of 5-fluorouracil (5-FU), theophylline (Th), and 6-mercaptopurine (6-MP), in addition to 5-FU and Th, using four models. Two series/parallel models have been developed that allow an aqueous-only path in parallel with a lipid-only path and with a lipid-aqueous series path for the permeation of solutes through skin: log J(M) = log ?1/[1/(aS(LIPID) 10(PhiMW)) + 1/(bS(AQ)/MW(1/2))] + cS(LIPID)10(PhiMW) + dS(AQ)/MW(1/2)? where a, b, c, and d are coefficients for flux through the lipid and aqueous portions of the series path, the lipid-only path, and the aqueous-only path, respectively, and Phi is the dependence of diffusivity in lipid on MW. In the first series/parallel model, S(LIPID) was predicted by S(IPM), and in the second, solvatochromic series/parallel model, S(LIPID) was predicted by S(IPM)(k MW + Omegai) where Omega(i) is the sum of the solvatochromic terms alpha, beta, pi, and R(2), and k is the coefficient for the dependence of partitioning on MW. Using the n = 41 solutions, the coefficients for the aqueous-only path were very small or not different from zero in the two series/parallel models, so only two-path series/parallel models were compared with the solvatochromic and transformed Potts-Guy models where a homogeneous barrier to permeation was assumed. For each model, one compound at a time was omitted from the data set and new parameter estimates were obtained for these 41-1 solutions and used to predict log J(M) for the omitted compound. The average errors of prediction of log J(M) (experimental log J(M) - predicted log J(M)) for the four models were 0.134 for the series/parallel (r(2) = 0.937), 0.127 for the solvatochromic series/parallel (r(2) = 0.967), 0.150 for the solvatochromic (r(2) = 0.950), and 0.134 log units for the transformed Potts-Guy model (r(2) = 0.944). Thus, the solvatochromic series/parallel model provides fit and predictive ability comparable to or slightly superior to previous models that assumed homogeneity of the diffusional barrier to flux in the rate-determining step, provides further theoretical support against the existence of a high capacity aqueous-only path, and provides further insight into the physicochemical properties that should be incorporated into solutes to optimize their flux. Using the solvatochromic series/parallel model, the parameter estimates for the n = 41 solution were used to calculate the flux of each compound through the two paths. For compounds with log partition coefficients (K(IPM:AQ)) of <0.8, permeation was mostly by the lipid-aqueous series path; for compounds with log K(IPM:AQ) >1.0, permeation was mostly by the lipid-only path; the lipid-aqueous series path exhibited the higher carrying capacity. (c) 2000 Wiley-Liss, Inc. PMID- 11015688 TI - Determination of average crystallite shape by X-ray diffraction and computational methods. AB - The objective of this work was to develop a method to estimate the average shape and habit of organic crystalline material using X-ray powder diffraction (XRPD), the single-crystal structure, and computational methods. It is proposed that the relative intensities of the peaks in an XRPD pattern from a sample exhibiting a "standard" preferred orientation correlates with the shape of the crystallites present. Models were developed to yield a quantitative "enhancement" factor for each face. The combined simple-forms morphology (CSM) of the material was then produced by indexing the observed faces and modifying the simulated Bravais Friedel-Donnay-Harker (BFDH) morphology. The average shape of crystallites can be estimated from the CSM by multiplying each face by its enhancement factor. Acetaminophen crystals in two different habits and ibuprofen crystallized from toluene were used. The predicted shapes closely resembled the average shapes observed with microscopy. Results suggested the average shapes of the organic crystalline materials can be estimated by XRPD and the computational simulation. The current limitations are the need to "index" the faces, the size of the crystallites, and the unknown impact of a polydisperse size distribution on the calculation. The method must be used within the limits described; however, it is the only method found that may be adapted to large, more representative sample sizes. The determination of the average morphology is often a "bottle neck" in elucidating other important behaviors of large quantities of crystalline powders used in pharmaceutical development and processing. PMID- 11015689 TI - Complexation of zolpidem with 2-hydroxypropyl-beta-, methyl-beta-, and 2 hydroxypropyl-gamma-cyclodextrin: effect on aqueous solubility, dissolution rate, and ataxic activity in rat. AB - The effect of some chemically modified cyclodextrins [namely, 2-hydroxypropyl beta-, methyl-beta-, and 2-hydroxypropyl-gamma-cyclodextrin (HP-beta-CD, Me-beta CD, and HP-gamma-CD, respectively)] on the aqueous solubility and dissolution rate of the hypnotic agent Zolpidem (ZP) was investigated. Solid complexes were prepared by freeze drying and characterized by infrared spectroscopy, X-ray powder diffraction, and differential scanning calorimetry. The solubility and dissolution rate of the drug were significantly improved by complexation with HP beta-CD or Me-beta-CD. The structure of the inclusion complex ZP-HP-beta-CD in CH(3)COOD/D(2)O was investigated by (1)H and (13)C NMR spectroscopy, including NOE measurements. These measurements revealing a weak interaction between the tolyl moiety of the guest molecule and the HP-beta-CD cavity. The ataxic activity in rat was also investigated and it was found that ZP-HP-beta-CD and ZP-Me-beta CD complexes showed almost 2-fold longer ataxic induction times than controls. PMID- 11015690 TI - Design of folic acid-conjugated nanoparticles for drug targeting. AB - The new concept developed in this study is the design of poly(ethylene glycol) (PEG)-coated biodegradable nanoparticles coupled to folic acid to target the folate-binding protein; this molecule is the soluble form of the folate receptor that is overexpressed on the surface of many tumoral cells. For this purpose, a novel copolymer, the poly[aminopoly(ethylene glycol)cyanoacrylate-co-hexadecyl cyanoacrylate] [poly(H(2)NPEGCA-co-HDCA)] was synthesized and characterized. Then nanoparticles were prepared by nanoprecipitation of the obtained copolymer, and their size, zeta potential, and surface hydrophobicity were investigated. Nanoparticles were then conjugated to the activated folic acid via PEG terminal amino groups and purified from unreacted products. Finally, the specific interaction between the conjugate folate-nanoparticles and the folate-binding protein was evaluated by surface plasmon resonance. This analysis confirmed a specific binding of the folate-nanoparticles to the folate-binding protein. This interaction did not occur with nonconjugated nanoparticles used as control. Thus, folate-linked nanoparticles represent a potential new drug carrier for tumor cell selective targeting. PMID- 11015691 TI - Structural characterization of three crystalline modifications of telmisartan by single crystal and high-resolution X-ray powder diffraction. AB - Three crystalline modifications (A, B, and C) of 4'-[[2-n-propyl-4-methyl-6-(1 methyl-benzimidazol-2-yl)benzi midazol-1-yl]methyl]biphenyl-2-carboxylic acid (INN name, telmisartan) have been detected and their crystal structures have been determined by single-crystal X-ray diffraction (pseudopolymorph C) and the method of simulated annealing from high-resolution X-ray powder diffraction data (polymorphs A and B). The compound is of interest because of its use as an angiotensin II receptor antagonist. Polymorph A crystallizes in space group P2(I)/c, Z = 4, with unit cell parameters a = 18.7798(3), b = 18.1043(2), and c = 8.00578(7) A, beta = 97.066(1) degrees, and V = 2701.31 A(3). Polymorph B crystallizes in space group P2(I)/a, Z = 4, with unit cell parameters a = 16.0646(5), b = 13.0909(3), and c = 13.3231(3) A, beta = 99.402(1) degrees, and V = 2764.2(1) A(3). The solvated form C crystallizes in space group C2/c, Z = 8, with unit cell parameters a = 30.990(5), b = 13.130(3), and c = 16.381(3) A, beta = 95.02(2) degrees, and V = 6639(2) A(3). For the structure solutions of polymorphs A and B, 13 degrees of freedom (3 translational, 3 orientational, 7 torsion angles) were determined in approximately 2 h of computer time, demonstrating that the crystal packing and the molecular conformation of medium sized (MW approximately 500) pharmaceutical compounds can now be solved quickly and routinely from high-resolution X-ray powder diffraction data. PMID- 11015692 TI - Effect of zinc binding and precipitation on structures of recombinant human growth hormone and nerve growth factor. AB - Metal-induced precipitation of protein therapeutics is being used and further developed as a processing step in protein formulation and may have utility in protein purification and bulk storage. In such processes, it is imperative that native protein structure is maintained and the metal complexation is reversible. In the current study, we investigated the effects of zinc-induced precipitation on recombinant human growth hormone (rhGH) and recombinant human nerve growth factor (rhNGF). On the addition of ethylenediaminetetraacetic acid (EDTA), the precipitates were dissolved, yielding complete recovery of native protein in both cases. Both proteins have specific metal binding sites and require specific molar ratios of zinc to protein to initiate precipitation (zinc:rhGH > 2:1; zinc:rhNGF > 18:1). Furthermore, the secondary structures of both proteins were unperturbed in soluble zinc complexes and zinc-induced precipitates, as measured by infrared and circular dichroism spectroscopies. The soluble zinc complex of rhGH had minor tertiary structural alterations, whereas zinc binding did not alter the tertiary structure of rhNGF. These studies indicated that metal-induced precipitation provides a method to maintain proteins in their native state in precipitates, which may be useful for purification, storage, and formulation. PMID- 11015694 TI - Physicochemical and crystallographic evidence for polymorphism of the racemic ethyl (2-chloromethyl-2,3-dihydro-5H-oxazolo [3, 2-a]pyrimidin-5-one)-6 carboxylate. AB - The various crystalline forms of an original bicyclic compound [ethyl (2 chloromethyl-2,3-dihydro-5H-oxazolo[3, 2-a]pyrimidin-5-one)-6-carboxylate); EOC] have been obtained and characterized by powder and single-crystal X-ray diffraction, differential scanning calorimetry (DSC), and infrared (IR) and Raman spectroscopy. At 4 degrees C in methanol, a monoclinic racemate (form II) crystallized from the racemic mixture, whereas at 20 degrees C, an orthorhombic racemate (form I) was isolated in trichloroethylene. By increasing the temperature, a solid-solid transition from the stable form II to the stable form I was observed with a Guinier-Simon camera. A I --> II transformation was observed at ambient temperature by DSC. PMID- 11015693 TI - Release characteristics of a short-chain fatty acid, n-butyric acid, from its beta-cyclodextrin ester conjugate in rat biological media. AB - 6(A)-O-(n-Butanoyl)-beta-cyclodextrin was prepared and its hydrolysis behavior in aqueous solutions and in rat intestinal fluids was investigated. Furthermore, the enzymatic hydrolyses of the n-butyric acid-beta-cyclodextrin conjugate using alpha-amylase and esterase were studied to gain insight into the release behavior of n-butyric acid from the conjugate. The hydrolysis of the conjugate proceeded according to a first-order kinetics in aqueous solution, and gave a V-shaped pH profile, indicating a specific acid-base-catalyzed hydrolysis at acidic and neutral-alkaline regions, respectively. The half-lives (t(1/2)) of the conjugate at pH 4.4, 6.8, and 7.4 at 37 degrees C were approximately 580, 43, and 6 days, respectively, indicating that the conjugate is stable in aqueous solution. No appreciable release of n-butyric acid from the conjugate was observed in the stomach and small intestinal contents of rats, or in the small and large intestinal homogenates of rats. On the other hand, a fast disappearance of the conjugate and an appearance of n-butyric acid were observed in the cecal and colonic contents of rats. The t(1/2) values of the disappearance were approximately 4, 1, and 6 h in 10 and 15% cecal contents and 10% colonic contents, respectively, and the appearance of n-butyric acid after 6 h was approximately 10% in the 15% cecal contents. Aspergillus oryzae alpha-amylase hydrolyzed the conjugate to small saccharide conjugates, such as the triose and maltose conjugates, but there was no appreciable release of n-butyric acid. The conjugate was less susceptible to carboxylic esterase (from porcine live), thus releasing no appreciable amounts of n-butyric acid. On the other hand, a fast release of n-butyric acid was observed when the esterase was employed after amylase hydrolysis, suggesting that two types of enzymes, sugar-degrading and ester-hydrolyzing enzymes, are necessary for the release of n-butyric acid from the conjugate in large intestinal contents. PMID- 11015695 TI - Megacystis-microcolon-intestinal hypoperistalsis syndrome: the difficulties with antenatal diagnosis. Case report and review of the literature. AB - Megacystis on antenatal scan in female fetuses is rare and has serious diagnostic implications. We report two cases of megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS) in female infants in whom antenatal scan abnormalities were identified, but the diagnosis not made until after delivery. MMIHS is a rare autosomal recessive condition which is usually lethal in the first year of life. Prenatal diagnosis is hampered by the lack of specific diagnostic findings on ultrasound and the absence of an identified genetic locus. The prenatal findings in MMIHS are reviewed and contrasted with those of other causes of lower abdominal masses on antenatal ultrasound. PMID- 11015696 TI - First trimester umbilical artery pulsatility index in fetuses presenting enlarged nuchal translucency. AB - The aim of this prospective study was to evaluate an umbilical artery pulsatility index (UAPI) in first trimester fetuses who present enlarged nuchal translucency (NT) measurements. UAPI was measured in 121 single fetuses with NT > or =95th centile, from 10+3 to 13+6 weeks (crown-rump length (CRL) > or =38 mm - < or =84 mm). In these fetuses there were 20 trisomy 21, and six other chromosomal abnormalities (three trisomy 18 and three monosomy X). Eighty-nine cases had normal karyotype and delivered a baby without evidence of congenital malformations. Five fetuses with normal karyotype assessed antenatally were excluded from the comparison, because of evidence of congenital malformation. A fetus with normal karyotype that was spontaneously miscarried at 14 weeks' after chorionic villus sampling was also excluded. UAPI of fetuses with enlarged NT was compared with those of 65 singleton fetuses with normal NT and normal karyotype, which were used to establish our terms of reference (5th centile, median and 95th centile). UAPI of 7/20 (35%) Down syndrome and 42/89 (47%) normal karyotype fetuses presenting enlarged NT were above the median, and respectively 2/20 (10%) and 14/89 (15.7%) were above 95th centile of normal NT and normal karyotype fetuses. No significant differences were demonstrated in the UAPI values amongst normal karyotype fetuses with normal NT or normal karyotype fetuses with an enlarged NT or trisomy 21 fetuses with an enlarged NT. PMID- 11015697 TI - How women deal with the results of serum screening for Down syndrome in the second trimester of pregnancy. AB - To gain insight into how pregnant women experience serum screening for Down syndrome, we sent questionnaires to two groups of relevant subjects in the north of the Netherlands. The questionnaires addressed the following issues: decision making process, knowledge and opinions. Questionnaire A was sent to women of 36 years of age and older (n=99) (group A) who were all 20 to 36 weeks pregnant at that time. In the Netherlands prenatal diagnosis is routinely available to these women. Questionnaire B was sent to women of younger than 36 years (n=69) (group B) who had received a screen-positive result and had subsequently undergone amniocentesis. About half of these women were still pregnant at that time. For these women, serum screening is only available on the basis of opting-in. The two questionnaires were largely identical. The response rates to questionnaires A and B were 82% and 91%, respectively. Group A (women of 36 years and older) considered that second trimester serum screening made a welcome contribution to the decision-making process about whether to undergo amniocentesis. Moreover, it reduced the amniocentesis rate considerably. The vast majority said they would apply for serum screening in a following pregnancy, but favoured the idea of first trimester screening. In group B (women of younger than 36 years), reassurance was the most commonly mentioned reason for undergoing serum screening. Almost all the women experienced some degree of anxiety when they were informed about the screen-positive result and 13% continued to be anxious, even after the favourable result of the amniocentesis. The majority of the respondents would also apply for serum screening in a following pregnancy and were of the opinion that this screening should be offered to all pregnant women in the Netherlands. PMID- 11015698 TI - Trends in live birth prevalence of down syndrome in the Northern Netherlands 1987 96: the impact of screening and prenatal diagnosis. AB - In the Northern Netherlands, we examined the live birth prevalence of Down syndrome (DS) and the impact of maternal serum screening (MSS) and prenatal cytogenetic diagnosis (PCD) during the period 1987-96. In this period the live birth prevalence, based on the maternal age distribution and the age specific risk of delivering a child with DS was expected to increase from 1.26 in 1987 to 1.62 in 1996. The introduction of MSS in 1991 made PCD available to women of all ages. Nevertheless, the utilization of PCD remained very stable. In 1991, 4.7% of pregnant women underwent a diagnostic test. In 1996 this percentage was 6.4%. As a result of MSS and PCD, the live birth prevalence of DS was 19% lower than expected (p<0.01). Despite utilization of PCD based on opting-in and a discouraging government policy regarding the offer of MSS, the percentage of DS cases detected by PCD increased from of 17% during the period 1987-90 to 27% in the period 1991-96 when MSS was available. The percentages have been corrected for spontaneous pregnancy loss. From a medical and financial point of view, MSS was the most cost-effective indication for PCD. However, the potential of reducing the birth prevalence of DS is limited by the low utilization of MSS and PCD by pregnant women. PMID- 11015699 TI - Numbers or words? A randomized controlled trial of presenting screen negative results to pregnant women. AB - The Objective of this study was to test the hypothesis that presenting risk information using numbers rather than words is a more effective way of communicating the residual risk inherent in a screen negative test result. We used a randomised controlled trial in a large UK teaching hospital. Two hundred and twenty pregnant women who received negative results on serum screening for Down syndrome participated. Presentation of screen negative test results were given either as a numerical probability (e.g. you have a 1:650 chance of having a baby with Down syndrome) or as a verbal probability (your chance of having a baby with Down syndrome is low). In both interventions the verbal anchor 'it is unlikely that your baby has Down syndrome' was used. Our aims were to measure the understanding of the residual risk in a screen negative result, and anxiety. Immediately after receipt of the results, 97% of those receiving their results in the form of a numerical probability and 91% of those receiving their results in the form of a verbal probability correctly understood that their baby probably did not have Down syndrome (95% CI for difference: 0% to 12%; p=0.04). All those who were incorrect believed that their baby definitely did not have Down syndrome. Subgroup analysis showed that this effect was confined to those with lower levels of education (i.e. those without a university degree), amongst whom understanding was poorer. There was no difference between intervention groups in understanding the results at four months. There were no differences between intervention groups in the levels of anxiety at one week or four months after receiving their results. In conclusion, communicating residual risks using numbers rather than words has a small beneficial effect of increasing awareness of residual risks without raising anxiety. Further work is needed to estimate the size of this effect in less well-informed and less highly educated populations. PMID- 11015700 TI - The Genoa experience of prenatal diagnosis in NF1. AB - Type 1 neurofibromatosis (NF1) is an autosomal dominant disorder with an incidence of about 1 in 3500 live births. Symptoms are highly variable from a few cafe-au-lait spots and axillary freckling to plexiform neurofibromas, optic gliomas, pseudarthrosis, and malignancy. Since disease causing mutations are dispersed throughout the gene, prenatal diagnosis is usually performed in familial cases by linkage analysis and rarely by direct characterization of the mutation. We have characterized 48 families and have performed four prenatal diagnoses. In three cases, the linkage analysis was carried out using informative markers. A direct approach using the protein truncation test (PTT) and sequencing was performed in one case in which a R1947X mutation was identified. The extreme variability of the phenotypic expression of the NF1 gene makes reproductive decisions in NF1 families very difficult, as molecular diagnosis cannot predict clinical expression of the disease. The psychological management of the couple is therefore difficult. In two of the three examined families the reproductive choices were not influenced by the specific manifestations of the disease in that family. PMID- 11015701 TI - How useful is the in vitro expansion of fetal CD34+ progenitor cells from maternal blood samples for diagnostic purposes? AB - Fetal cells present in the maternal circulation are a potential source of fetal DNA that can be used for the development of a prenatal diagnostic test. Since their numbers are very low, amplification of fetal cells has been discussed for a long time. So far, most studies have focused on culturing fetal erythroid cells. In this study, we evaluated whether limiting numbers of fetal haemopoietic progenitor cells present in an excess of maternal cells were able to overgrow the maternal component. Therefore, we used a model system in which limiting numbers of male CD34+ umbilical cord blood cells were diluted in 400 000 female CD34+ peripheral blood cells. The number of XY positive cells derived from umbilical cord blood was determined using two-colour in situ hybridization with X and Y chromosomal probes. We demonstrated a 1500-fold relative expansion of male umbilical cord blood cells over the peripheral blood component after three weeks of liquid culture, which also corresponded to the extent of expansion of CD34+ cells derived from 20-week fetal blood. However, application of the same culture protocol to maternal blood samples obtained at 7-16 weeks of gestation showed no preferential growth of fetal haemopoietic progenitor cells. This study, therefore, suggests that fetal primitive haemopoietic progenitor cells do either not circulate in maternal blood before 16 weeks of gestation, or require different combinations/concentrations of cytokines for their in vitro expansion. PMID- 11015702 TI - Determination of enzyme activities for prenatal diagnosis of respiratory chain deficiency. AB - Genetic counselling and prenatal diagnosis are major issues of mitochondrial respiratory chain deficiency, especially as these conditions are largely untreatable. In the absence of known mitochondrial or nuclear gene mutations, measurement of respiratory chain enzyme activities represents the only possibility to prevent recurrence of the disease in affected families. We carried out enzymatic prenatal diagnosis in 21 pregnancies from 10 unrelated couples using uncultured choriocytes and/or amniocytes. Twelve babies were born and are healthy, seven pregnancies were discontinued early on because of an enzyme deficiency detected prenatally. In two cases, a fetus which appeared normal after early and/or late prenatal diagnosis, turned out to be affected. We conclude that a deficient enzyme activity is indicative of recurrence, but a normal result at 10 weeks of gestation does not give conclusive evidence as to the outcome of the pregnancy. We therefore suggest the following procedure: (1) a choriocentesis or an amniocentesis in early pregnancy when the proband expresses the disease in cultured skin fibroblasts; (2) a second amniocentesis at 28 weeks' gestation should be offered to avoid false negative results due to a possible late expression of the disease, in combination with: (3) a careful and repeated ultrasound survey for detection of growth failure in the third trimester; (4) prenatal diagnosis should not be performed in case of late onset clinical symptoms in the proband; and (5) parents should be aware of the possibility of false negative results. Prenatal diagnosis should not be proposed for a complex I deficiency as this enzyme activity cannot be accurately measured in fetal cells. PMID- 11015703 TI - Mid-trimester beta-hCG levels incorporated in a multifactorial model for the prediction of severe pre-eclampsia. AB - Pre-eclampsia remains a major cause of perinatal morbidity and mortality worldwide. Proposed predicting tests for early detection of pregnant women destined to develop pre-eclampsia remain unsatisfactory. The aim of this study was to investigate the clinical utility of combining mid-trimester maternal serum beta-human chorionic gonadotrophin (MShCG) levels with selected clinical determining factors as a multifactorial predictive test for pre-eclampsia. Thirty nine cases with mild pre-eclampsia and 56 with severe pre-eclampsia were recruited as the study groups. Normotensive women (957) were enrolled as controls. Potential determining risk factors for severe pre-eclampsia were selected using a multiple logistic regression to build various combined prediction models. A receiver-operator characteristic curve was employed to assess the performance of each prediction test for pre-eclampsia. The prediction efficacy of each test was examined by the area under the curve (AUC). Our data show that mid-trimester MShCG levels significantly correlated with severity of pre-eclampsia (Spearman rank correlation coefficient=0.195, p<0.001). Women with mild pre-eclampsia had a 2.61-times greater chance, while women with severe pre eclampsia had a 6.13-times greater chance of having MShCG exceeding 2.0 multiples of the median than did women with a normal pregnancy. A combined prediction model composed of MShCG levels, body mass index (BMI), parity, and age as a predictive test for severe pre-eclampsia was superior to MShCG levels alone (AUC 0.765 versus 0.648). The integrated multifactorial model could identify women at risk early on for developing severe pre-eclampsia, with a sensitivity of 70% and a specificity of 71%. Thus, we demonstrate a potentially effective and convenient method by which women at risk for developing severe pre-eclampsia can be identified early, based on a multifactorial predictive model composed of midtrimester MShCG levels, BMI, parity, and age. PMID- 11015704 TI - Prenatal molecular diagnosis of L1-spectrum disorders. PMID- 11015705 TI - Antepartum findings in fetal protein C deficiency. AB - A pregnancy with fetal homozygous protein C deficiency was complicated in the third trimester by fetal ventriculomegaly, intraorbital thrombosis and placental infarcts, which could be imaged by combined use of ultrasonography and MRI. PMID- 11015706 TI - Prenatal diagnosis and genetic analysis of double trisomy 48,XXX,+18. AB - Prenatal diagnosis of simultaneous occurrence of double trisomy involving chromosomes 18 and X is extremely rare. We report on the prenatal diagnosis, genetic analysis and clinical manifestations of a fetus with both trisomy 18 and trisomy X. A 26-year-old, para 1 woman was referred for genetic counselling at 36 weeks' gestation with the sonographic findings of intrauterine growth retardation (IUGR), polyhydramnios, ventricular septal defect, and an enlarged cisterna magna. Both cordocentesis and amniocentesis revealed a consistent karyotype of 48,XXX,+18. Quantitative fluorescent polymerase chain reaction using polymorphic small tandem repeat markers specific for chromosomes 18 and X rapidly determined that both aneuploidies arose as a result of non-disjunction in maternal meiosis II. Our case shows that two non-disjunction events can occur not only in the same parent, but also in the same cell division. Our case also shows that double trisomy, 48,XXX,+18, can demonstrate an enlarged cisterna magna, IUGR and polyhydramnios in prenatal ultrasound. PMID- 11015707 TI - Prenatal diagnosis and genetic analysis of X chromosome polysomy 49, XXXXY. AB - We report on the prenatal diagnosis, genetic analysis and clinical manifestations of a 49,XXXXY fetus. A 31-year-old, primigravida woman was referred for genetic counselling at 17 weeks' gestation with the sonographic findings of intrauterine growth retardation, generalized oedema, a large septated cystic hygroma colli measuring 5x4 cm, and abnormal posturing of the lower extremities. Quantitative fluorescent polymerase chain reaction (QF-PCR) with small tandem repeat (STR) markers specific for chromosome X and a pentanucleotide marker X22 for the Xq/Yq pseudoautosomal region PAR2 rapidly detected the X-chromosome polysomy from amniotic fluid cells. This abnormality appeared to arise from successive non disjunction during maternal meiosis I and meiosis II. Cytogenetic analysis revealed a karyotype of 49,XXXXY. Our case shows that a 49,XXXXY fetus in the second trimester may demonstrate hydrops fetalis and a large septated cystic hygroma colli by prenatal ultrasound. Our case also shows that QF-PCR assays with sex chromosome specific STR markers provide rapid prenatal diagnosis of numerical sex chromosome aneuploidy as well as its genetic cause in fetal cystic hygroma. PMID- 11015708 TI - A new approach to prenatal treatment of extralobar pulmonary sequestration. AB - Fetal hydrothorax and hydrops is a frequent complication of extralobar pulmonary sequestration which is associated with a high perinatal mortality and severe respiratory insufficiency in the newborn. In a 27-week-old fetus with this condition, injection of 1 ml of pure alcohol and pleuro-amniotic shunting achieved resolution of hydrops. The pregnancy progressed to term and a healthy neonate was delivered who did not require postnatal surgery. PMID- 11015709 TI - Prenatal molecular diagnosis of glutaric aciduria type I by direct mutation analysis. AB - Various biochemical strategies are followed for the prenatal diagnosis of glutaric aciduria type I (GA I). However, since the description of patients with normal excretion of glutarate and significant residual activity, the difficulties of prenatal biochemical diagnosis are obvious. The characterization of the glutaryl-CoA dehydrogenase (GCDH) gene has allowed us to develop a single strand conformation polymorphism (SSCP) screening method, followed by direct sequencing, to identify the disease causing mutations in patients with GA I. Here we report the first prenatal diagnoses based on DNA analysis in chorionic villi biopsy or cultured amniotic fluid cells in three families at risk for GA I. Our results show that this strategy provides a fast and reliable method for prenatal diagnosis. In addition we report two new mutations (1209-1210ins G and R161W) in the GCDH gene that occurred at hypermutable loci. PMID- 11015710 TI - Prenatal diagnosis of glycogen storage disease type 1b using denaturing high performance liquid chromatography. AB - Glycogen storage disease type 1b (GSD1b) is an autosomal recessive inborn error of metabolism caused by deficiency of glucose-6-phosphate translocase (G6PT1). Current laboratory diagnosis for GSD1b is established by a functional enzyme assay of glucose-6-phosphatase in both fresh and detergent-treated liver homogenates. This procedure requires liver biopsy and is impractical for routine prenatal diagnosis owing to the high morbidity of fetal liver biopsy. Recently, the gene for GSD1b has been cloned and the prevalent mutations in different ethnic groups have been determined. In this study, prenatal molecular diagnosis was performed for a Chinese family in which a previous child was born homozygous for the G149E mutation. We detected genomic sequence variants by heteroduplex formation, followed by denaturing high performance liquid chromatography (DHPLC). With this method, post-PCR analysis was shortened to 7 min. In the case we analysed, PCR products amplified from the fetal DNA yielded a single peak in the chromatogram, indicating a homozygous state in the fetus. When wild-type PCR products were mixed with fetal PCR products, two peaks were observed, indicating that the fetus was homozygous for the parental (G149E) mutation. Sequencing results confirmed this diagnosis. As a result, the pregnancy was terminated and the diagnosis was confirmed on DNA analysis of the aborted fetus. We show here that DNA mutation analysis can be used in the prenatal diagnosis of GSD1b and that DHPLC promises to be a robust technique for this and other prenatal molecular diagnoses. PMID- 11015711 TI - Prenatal diagnosis of anophthalmos with limb-body wall complex. PMID- 11015712 TI - Maternal serum total hCG and free beta-hCG in the first trimester from trisomy 21 pregnancies PMID- 11015713 TI - Maternal serum total hCG and free beta-hCG in the first trimester from trisomy 21 pregnancies. PMID- 11015714 TI - Current awareness. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of prenatal diagnosis. Each bibliography is divided into 17 sections: 1 Books, Reviews & Symposia; 2 General Interest; 3 Normal Fetal Development; 4 Gametogenesis and Pre-implantation Diagnosis; 5 First Trimester Diagnosis; 6 Second Trimester Diagnosis; 7 Fetal Diagnosis by Ultrasound and Other Imaging; 8 Maternal Screening; 9 Screening for Carriers of Genetic Abnormality; 10 Technological Developments; 11 Confined Placental Mosaicism and Uniparental Disomy; 12 Molecular Cytogenetics; 13 Fetal Cells in Maternal Circulation; 14 Fetal Therapy; 15 Psychosocial Aspects; 16 Epidemiology and Environmental Factors; 17 Developmental Pathology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted PMID- 11015715 TI - Micrognathozoa: a new class with complicated jaws like those of Rotifera and Gnathostomulida. AB - A new microscopic aschelminth-like animal, Limnognathia maerski nov. gen. et sp., is described from a cold spring at Disko Island, West Greenland, and assigned to Micrognathozoa nov. class. It has a complex of jaws in its pharynx, and the ultrastructure of the main jaws is similar to that of the jaws of advanced scleroperalian gnathostomulids. However, other jaw elements appear also to have characteristics of the trophi of Rotifera. Jaw-like structures are found in other protostome taxa as well-for instance, in proboscises of kalyptorhynch platyhelminths, in dorvilleid polychaetes and aplacophoran mollusks-but studies of their ultrastructure show that none of these jaws is homologous with jaws found in Gnathostomulida, Rotifera, and Micrognathozoa. The latter three groups have recently been joined into the monophylum Gnathifera Ahlrichs, 1995, an interpretation supported by the presence of jaw elements with cuticular rods with osmiophilic cores in all three groups. Such tubular structures are found in the fulcrum of all Rotifera and in several cuticular sclerites of both Gnathostomulida and Micrognathozoa. The gross morphology of the pharyngeal apparatus is similar in the three groups. It consists of a ventral pharyngeal bulb and a dorsal pharyngeal lumen. The absence of pharyngeal ciliation cannot be used as an autapomorphy in the ground pattern of the Gnathifera because the Micrognathozoa has the plesiomorphic alternative with a ciliated pharyngeal epithelium. The body of Limnognathia maerski nov. gen. et sp. consists of a head, thorax, and abdomen. The dorsal and lateral epidermis have plates formed by an intracellular matrix, as in Rotifera and Acanthocephala; however, the epidermis is not syncytial. The ventral epidermis lacks internal plates, but has a cuticular oral plate without ciliary structures. Two ventral rows of multiciliated cells form a locomotory organ. These ciliated cells resemble the ciliophores present in some interstitial annelids. An adhesive ciliated pad is located ventrally close to a caudal plate. As in many marine interstitial animals e.g., gnathostomulids, gastrotrichs, and polychaetes-a special form of tactile bristles or sensoria is found on the body. Two pairs of protonephridia with unicellular terminal cells are found in the trunk; this unicellular condition may be the plesiomorphic condition in Bilateria. Only specimens with the female reproductive system have been found, indicating that all adult animals are parthenogenetic females. We suggest that 1) jaws of Gnathostomulida, Rotifera, and the new taxon, Micrognathozoa, are homologous structures; 2) Rotifera (including Acanthocephala) and the new group might be sister groups, while Gnathostomulida could be the sister-group to this assemblage; and 3) the similarities to certain gastrotrichs and interstitial polychaetes are convergent. PMID- 11015716 TI - Decription of Limnognathia maerski. PMID- 11015717 TI - Catecholamine sensitivity in the rat femoral artery after microvascular anastomosis. AB - Tissue can demonstrate vasospastic instability after microvascular anastomosis. This study investigates the in vitro effect of increasing concentrations of phenylephrine on the rat femoral artery after microvascular anastomosis. Bilateral groin flaps based on the inferior epigastric artery were raised on 55 Wistar male rats. On the test side, a microvascular anastomosis was performed, but not on the control side. On days 2-12 postoperatively, the rats were sacrificed and the femoral arteries harvested and suspended in increasing concentrations of phenylephrine. The vascular tone on the test and control sides were recorded and compared. Increased sensitivity was found on the test side as compared with the control side (P = 0.000). This supersensitivity to phenylephrine was blocked by the addition of phentolamine and it is believed to be the result of sympathetic denervation, which occurs when the sympathetic fibers are cut during the harvesting of the flap. The resulting vascular instability is believed to contribute to flap failure. PMID- 11015718 TI - Microvascular surgery in a bloodless field. AB - Although tourniquets play an integral role in extremity surgery, no clear guidelines exist for the use of tourniquets in microsurgery. We undertook a study in 12 healthy volunteers to better understand the coagulation properties of blood distal to an inflated tourniquet. At a 15-min inflation time, blood distal to an inflated tourniquet clots faster than blood taken from the opposite arm after addition of exogenous thrombin (12.5 s vs 17.5 s, P < 0.0001). Neither fibrinopeptide A (FPA) levels nor tissue plasminogen activator (tPA) levels were different from those of controls. Tissue factor pathway inhibitor (TFPI), an endogenous local anticoagulant, was slightly but significantly elevated in tourniquet blood. Although much remains to be understood, we believe that microvascular surgery in a bloodless field is safe and efficacious. Nine patients are presented who successfully underwent microvascular surgery in a bloodless field, using various types of extremity tourniquets. PMID- 11015719 TI - Method for morphometric analysis of axons in experimental peripheral nerve reconstruction. AB - A new method for morphometric analysis of axons in experimental peripheral nerve reconstruction is presented. Twelve adult female rabbits were used. In nine animals the saphenous nerve was transected and stitched epineurially. Three animals functioned as control. After 3, 6, and 12 months, the nerves were harvested, fixed in Kryofix and embedded in Histowax. Transverse sections of 6 microm were cut, immunohistochemically stained for NF 90, and counterstained by Sirius Red. Quantification of nerve fibers in cross sections was performed by using a confocal laser scanning microscope (CLSM), and the images were stored digitally. Data analyzing was performed by the Optimas program (5.2). Calculations were done with Microsoft Excel. The total number of axons, the mean axon diameter and the percentage axon area/fascicle area were evaluated statistically. This method for morphologic analysis provides automatically complete registration of axons and so different methods of experimental nerve reconstruction can be compared in a fast and reliable way. PMID- 11015720 TI - Reconstruction after resection of tumors around the knee: role of the free vascularized fibular graft. AB - We present our experience with reconstruction after resection of tumors around the knee, using free vascularized fibular grafting. The study included 23 patients. The lower femur was involved in 17 cases, the upper tibia in 6. The cases included giant cell tumor of the lower femur (2 patients), giant cell tumor of the upper tibia (1 patient), malignant fibrous histiocytoma of the lower femur (1 patient), parosteal osteosarcoma (1 patient), and periosteal osteosarcoma (1 patient). The remaining patients suffered from conventional osteogenic sarcomas. The size of the defect ranged from 12 to 16 cm in length. Skin flap necrosis after tumor resection was the most common complication encountered. Other complications included peroneal nerve involvement in one case and rupture of the arterial anastomosis in another. All transferred fibulas progressed to union within 7-9 months. Union time of both upper and lower ends of the fibula and time of appearance of periosteal reaction were identical. In evaluating periosteal hypertrophy of the fibula, the hypertrophy (de Boer) index (de Boer HD, Wood MB, J Bone Joint Surg 1989;71B:374-378) proved unreliable. False positive results are obtained, when callus formation around the lower end of the femur is far more abundant than at the upper end of the fibula. For this reason, we introduced the graft index. The latter is the ratio between the diameter of the graft at its thinnest portion at latest follow-up to its diameter on the day of operation, as calculated on plain radiographs. Two of the viable fibulas developed stress fractures after plate removal. Functional and quality-of -life results were satisfactory. It is concluded that the free vascularized fibular graft remains a valuable reconstruction option after the resection of tumors around the knee, provided certain precautions are followed. First, before closure of the wound, the skin flaps should be assessed for their viability. Necrotic parts should be excised. Stable fixation is a necessary prerequisite at the time of operation. Removal of the fixation device should not be guided by union or periosteal hypertrophy, but by true widening of the medullary canal. PMID- 11015721 TI - Vascular complications of native arteriovenous fistulas for hemodialysis: role of microsurgery. AB - This article describes the treatment of 56 early and late vascular complications of native arteriovenous fistula (AVF) in patients with end-stage renal disease, between January 1987 and January 1999. Twenty-six were arteriovenous aneurysms and 2 pseudoaneurysms; 25 were thromboses. We also observed two cases of periarteritis associated with systemic lupus erythematosus (SLE) and one complex iatrogenic lesion caused by an attempt at percutaneous restoration of potency. We applied microsurgical principles, instruments, and techniques. It was possible to restore a vascular access at the original site, using the same vessels in 45 cases (80.4%). In 10.7% of cases, we were able to rescue the original AVF by microsurgical revision. A new vascular access had to be created proximally in the same limb or in the contralateral forearm in 11 cases of aneurysms (19.6% of the total, 42% of the aneurysms). After a minimum follow-up of 8 months, a total of four patients had to be reoperated for further complications (7.2%). Our data support the idea that microsurgical treatment of vascular complications of native AVF is highly successful compared with results obtained by conventional surgery and, in a defined subgroup of patients, permits salvage of the fistula. PMID- 11015722 TI - Cellular activity of resident macrophages during Wallerian degeneration. AB - The resident macrophages have been accepted as an important component of the peripheral nervous system as Schwann cells. To elucidate their role during Wallerian degeneration without interference from extrinsic hematogenous macrophages, we designed a culture system to investigate the behavior of resident macrophages in vitro. A total of 75 adult male Lewis rats were used; 2. 5-cm length sciatic nerve explants were harvested. There were three groups. In the culture groups, the nerve explants were incubated in Dulbecco's modified Eagle's medium (DMEM) only or in DMEM supplemented with 2 microm forskolin and 10 microg/ml pituitary extract (mitogenic medium for Schwann cells). In vivo predegenerated nerves and normal nerves were used as the positive and negative controls, respectively. The observation periods extended to 3 weeks. Hematoxylin and eosin (H&E) stain was employed to estimate overall cell number in nerve explants. Macrophages were labeled with ED1; S-100 immunostaining was used to evaluate the presence of Schwann cells during Wallerian degeneration. Trichrome stain and toluidine blue stain were used to visualize the fate of myelin. In the culture groups, the number of resident macrophages increased continuously, although there were significantly fewer resident macrophages than hematogenous macrophages after 3 days of Wallerian degeneration (P < 0.01). Morphologically, resident macrophages contained densely small ED1-positive granules within their cytoplasm, even at later stages of observation, whereas hematogenous macrophages contained typical large ED1-positive foam vacuoles characteristic of their mature phagocytic ability. The cellular activity of Schwann cells was well preserved in the mitogenic medium; however, myelin removal was not significantly enhanced as compared with the DMEM groups (P > 0.05). The clearance of myelin debris was shown to be incomplete in culture groups as compared with the complete removal of myelin debris in the in vivo groups. Resident macrophages were actively involved in Wallerian degeneration, but their phagocytic and proliferation ability was limited. Schwann cells played an adjunctive role during the removal of myelin debris. PMID- 11015723 TI - Flap transfers for the treatment of perichondrial ring injuries with soft tissue defects. AB - Seven cases (six fresh) of perichondrial ring injury with skin defects were treated using flap transfers. The study included four boys and three girls ranging in age from 2 to 9 years (average 6). They were followed up for an average of 8 years and 10 months. The period from injury to flap coverage was 8 12 days, with an average of 10 days in the fresh cases. Fracture was noted in four cases, with one an epiphyseal fracture. Peroneal flaps were transferred in four cases, latissimus dorsi myocutaneous flaps in two, and gastrocnemius muscle flap in one. Six flaps survived perfectly, and one failed due to venous thrombosis. This latter case was treated with a cross leg flap. Postoperative radiographic assessments confirmed partial growth plate arrest in the chronic case, but all the fresh cases had no postoperative growth disturbance. Flap coverage, for perichondrial ring injuries with wide skin defects, is a useful method not only for skin coverage, but for the prevention of growth disturbances as well. PMID- 11015724 TI - Schizosaccharomyces pombe gmd3(+)/alg11(+) is a functional homologue of Saccharomyces cerevisiae ALG11 which is involved in N-linked oligosaccharide synthesis. AB - The oligosaccharide of glycoproteins in the fission yeast Schizosaccharomyces pombe is unique in containing galactose. We isolated four mutants that had reduced amounts of galactose residues on their cell surface glycoproteins by fluorescence-activated cell sorter. The isolated four recessive mutants, gmd1 to gmd4, showed a defect in glycosylation of acid phosphatase, a cell surface glycoprotein. In gmd3 mutant cells, the amounts of both mannose and galactose residues were decreased on the cell surface galactomannoproteins, suggesting an underglycosylation of galactomannoproteins. The gmd3(+) gene encodes a protein that has significant similarity with Saccharomyces cerevisiae Alg11p and is likely to be involved in N-linked core oligosaccharide synthesis. ALG11 suppressed the gmd3 mutation, indicating that gmd3(+) gene is a functional homologue of the ALG11 gene. We therefore designated gmd3(+) as alg11(+). PMID- 11015725 TI - Purification, molecular and kinetic characterization of phosphofructokinase-1 from the yeast Schizosaccharomyces pombe: evidence for an unusual subunit composition. AB - Phosphofructokinase-1 (Pfk-1) from Schizosaccharomyces pombe was purified by 54 fold enrichment to homogeneity elaborating the following steps: (a) Disruption of the cells with glass beads; (b) fractionated precipitation with polyethylene glycol 6000; (c) affinity chromatography on Cibacron-Blue F3G-A-Sephadex G 100; (d) ion exchange chromatography on Resource Q. The native enzyme exhibits a mass of 790+/-30 kDa, as detected by sedimentation equilibrium measurements. The apparent sedimentation coefficient was found to be s(20,c)=20.2+/-0.3 S. No significant dependence of the s-value on the protein concentration was observed in the range 0. 07-0.7 mg/ml. Polyacrylamide gel electrophoresis in presence of sodium dodecyl sulphate and MALDI-TOF spectra showed that the enzyme is composed of subunits of identical size of 100+/-5 kDa, forming an octameric structure. The N-terminus of the enzyme was found to be blocked. Sequences of tryptic and chymotryptic peptides of the subunit coincide with the proposed amino acid sequence as deduced from the gene from the EMBL library. The Pfk-1 coding sequence of S. pombe was transformed into a Pfk-1 double deletion mutants of Saccharomyces cerevisiae resulting in glucose-positive cells with enzyme activity in the crude cell extract. The kinetic analysis revealed less cooperativity to fructose 6-phosphate (n(H)=1.6) and less inhibition by ATP as compared to the enzyme from baker's yeast. Fructose 2,6-bisphosphate (in micromolar range) and AMP (in millimolar range) were found to overcome ATP inhibition and to increase the affinity to fructose 6-phosphate. PMID- 11015726 TI - Alcohol acetyltransferases and the significance of ester synthesis in yeast. AB - This paper reviews our current knowledge of yeast alcohol acyltransferases. Much of this information has been gathered over the past 10 years through the application of powerful yeast molecular biology techniques. Evidence from gene disruption and expression analysis of members of the alcohol acyltransferase (ATF) gene family indicates that different ester synthases are involved in the synthesis of esters during alcoholic fermentation. The natural physiological rationale behind these enzyme activities remains unclear. However, it is believed that these enzymes may be involved in very different functions, including cellular fatty acid homeostasis and detoxification mechanisms. Insights into the regulation of yeast ester synthesis by oxygen and unsaturated fatty acids have contributed to our understanding of the general mechanisms of gene regulation. In particular, control mechanisms that underpin the oxygen-mediated regulation of ATF1 gene transcription appear to be closely linked to those involved in the regulation of fatty acid metabolism. Data pertaining to the regulation of ATF1 gene transcription have been integrated into a working model for future research. PMID- 11015727 TI - The yeast Ura2 protein that catalyses the first two steps of pyrimidines biosynthesis accumulates not in the nucleus but in the cytoplasm, as shown by immunocytochemistry and Ura2-green fluorescent protein mapping. AB - The Ura2 multidomain protein catalyses the first two steps of pyrimidines biosynthesis in Saccharomyces cerevisiae. It consists of a 240 kDa polypeptide which contains carbamyl phosphate synthetase and aspartate transcarbamylase domains. The Ura2 protein was believed to be nucleoplasmic, since one of the aspartate transcarbamylase reaction products, monophosphate, was reported to be precipitated by lead ions inside nuclei. However, this ultracytochemical approach was recently shown to give artifactual lead polyphosphate precipitates, and the use of cerium instead of lead failed to reveal this nucleoplasmic localization. Ura2 localization has therefore been undertaken by means of three alternative approaches based on the detection of the protein itself: (a) indirect immunofluorescence of yeast protoplasts; (b) immunogold labelling of ultrathin sections of embedded yeast cells (both approaches using affinity purified primary antibodies directed against the 240 kDa Ura2 polypeptide chain, or against a 22 residue peptide specific of the carbamyl phosphate synthetase domain); and (c) direct fluorescence of cells expressing an Ura2-green fluorescent protein hybrid. All three approaches localize the bulk of Ura2 to the cytoplasm, whereas the signals associated with the nucleus, mitochondria or vacuoles are close to or at the background level. PMID- 11015728 TI - Destabilized green fluorescent protein for monitoring dynamic changes in yeast gene expression with flow cytometry. AB - Green fluorescent protein (GFP) has many advantages as a reporter molecule, but its stability makes it unsuitable for monitoring dynamic changes in gene expression, among other applications. Destabilized GFPs have been developed for bacterial and mammalian systems to counter this problem. Here, we extend such advances to the yeast model. We fused the PEST-rich 178 carboxyl-terminal residues of the G(1) cyclin Cln2 to the C terminus of yEGFP3 (a yeast- and FACS optimized GFP variant), creating yEGFP3-Cln2(PEST). We tested the hybrid protein after integrating modules harbouring the yEGFP3 or yEGFP3-CLN2(PEST) ORFs into the Saccharomyces cerevisiae genome. yEGFP3- Cln2(PEST) had a markedly shorter half-life (t(1/2)) than yEGFP3; inhibition of protein synthesis with cycloheximide lead to a rapid decline in GFP content and fluorescence (t(1/2) approximately 30 min) in cells expressing yEGFP3-Cln2(PEST), whereas these parameters were quite stable in yEGFP3-expressing cells (t(1/2) approximately 7 h). We placed yEGFP3-CLN2(PEST) under the control of the CUP1 promoter, which is induced only transiently by copper. This transience was readily discernible with yEGFP3-Cln2(PEST), whereas yEGFP3 reported only on CUP1 switch-on, albeit more slowly than yEGFP3-Cln2(PEST). Cell cycle-regulated transcriptional activation/inactivation of the CLN2 promoter was also discernible with yEGFP3- Cln2(PEST), using cultures that were previously synchronized with nocodazole. In comparison to CLN2, expression from the ACT1 promoter was stable after release from nocodazole. We also applied a novel flow-cytometric technique for cell cycle analysis with asynchronous cultures. The marked periodicities of CLN2 and CLB2 (mitotic cyclin) transcription were readily evident from cellular yEGFP3 Cln2(PEST) levels with this non-perturbing approach. The results represent the first reported successful destabilization of a yeast-GFP. This new construct expands the range of GFP applications open to yeast workers. PMID- 11015729 TI - Simultaneous overexpression of enzymes of the lower part of glycolysis can enhance the fermentative capacity of Saccharomyces cerevisiae. AB - Recombinant S. cerevisiae strains, with elevated levels of the enzymes of lower glycolysis (glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate mutase, phosphoglycerate kinase, enolase, pyruvate kinase, pyruvate decarboxylase and alcohol dehydrogenase) were physiologically characterized. During growth on glucose the enzyme levels in the recombinant strains (YHM4 and YHM7) were 1.1-3.4 fold higher than in the host strain (CEN.PK.K45). The recombinant strains were grown in aerobic or anaerobic batch cultures on glucose or a mixture of glucose and galactose. The specific ethanol production rates in the recombinant strains were the same as for the host strain and the physiological behaviour of the recombinant strains and the host strain was similar. When the cellular demand for ATP was increased by means of glucose pulses (final concentrations of 3.9 g/l or 2.0 g/l, respectively) to aerobic chemostat cultures maintained at a dilution rate of 0.08/h, the specific carbon dioxide production rate (qCO(2)) of CEN.PK.K45 accelerated at 6x10(-3) mmol/g/min(2) during the first 15 min, whereas during the same time period the qCO(2) of YHM7 accelerated twice as fast at 12x10(-3) mmol/g/min(2), indicating a higher fermentative capacity in the recombinant strain. PMID- 11015730 TI - Diversity of the HO gene encoding an endonuclease for mating-type conversion in the bottom fermenting yeast Saccharomyces pastorianus. AB - Two types of HO gene were cloned, sequenced and characterized from the bottom fermenting yeast Saccharomyces pastorianus. The HO gene present on the 1500 kb chromosome was designated Sc-HO (S. cerevisiae-type HO), because the nucleotide sequence of its promoter region and the open reading frame (ORF) was almost identical to that of the S. cerevisiae laboratory strain HO gene (Lab-HO). The other HO gene, designated Lg-HO (Lager-fermenting-yeast specific HO), showed 64% and 83% homology with the promoter and ORF of the Lab-HO at the nucleotide sequence level, respectively, and was located on the 1100 kb chromosome. Analysis of the 4 kb DNA fragment amplified from S. bayanus type strain indicated that the nucleotide sequence of S. bayanus-HO is almost identical to that of the Lg-HO. The SSB1 gene located downstream of the HO gene in S. cerevisiae was also found in the 3' distal region of the Sc-HO, Lg-HO and S. bayanus HO genes. These results showed that the genetic arrangement around the HO loci both of S. pastorianus and S. bayanus is identical to S. cerevisiae. Southern analysis has revealed that Saccharomyces sensu stricto contain four types of HO genes; S. paradoxus-type HO, the Sc-HO, the Lg-HO and S. uvarum-type HO genes. This HO gene diversity provides useful information for the classification of strains belonging to Saccharomyces sensu stricto. The S. pastorianus Sc-HO, Lg-HO and S. bayanus-HO Accession Nos in the DDBJ Nucleotide Sequence Database are AB027449, AB027450 and AB027451, respectively. PMID- 11015731 TI - Construction of fission yeast vectors with a novel selection strategy that allows their use in wild-type fission yeasts. AB - Novel vectors that use the Pichia pastoris INO1 gene as a selectable marker and exploit the natural inositol auxotrophy of the fission yeast are described. These plasmids also contained other features desirable in a plasmid cloning vector. These plasmids were evaluated in other species of Schizosaccharomyces and found to replicate autonomously in another variety of S. pombe, S. pombe var. malidevorans. These plasmids can be used for transformation of any wild-type S. pombe strain without the need for selection by induced auxotrophic mutations, or by selection by drug resistance markers, and should greatly assist genetic and molecular manipulations in these yeasts. PMID- 11015732 TI - Current awareness on yeast. PMID- 11015739 TI - Herpesviruses as unrecognised components of the pathogenesis of chronic diseases. PMID- 11015740 TI - Herpes simplex virus as a cause of Bell's palsy. 1972. PMID- 11015741 TI - Transient virus infection and multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic, demyelinating disease of the CNS in which autoimmunity to myelin plays a role in pathogenesis. The epidemiology of MS indicates that it may be triggered by a virus infection before the age of adolescence, but attempts to associate a specific virus with MS have produced equivocal results. Many studies of the aetiology of MS have postulated that a persistent virus infection is involved, but transient virus infection may provide a plausible alternative mechanism that could explain many of the inconsistencies in MS research. The most studied animal model of MS is chronic relapsing experimental autoimmune encephalomyelitis (CREAE), which is induced in susceptible animals following injection of myelin components. While CREAE cannot provide information on the initiating factor for MS, it may mimic disease processes occurring after an initial trigger that may involve transient virus infection. The disease process may comprise separate triggering and relapse phases. The triggering phase may involve sensitisation to myelin antigens as a result of damage to oligodendrocytes or molecular mimicry. The relapse phase could be similar to CREAE, or alternatively relapses may be induced by further transient virus infections which may not involve infection of the CNS, but which may involve the recrudescence of anti-myelin autoimmunity. Although current vaccines have a high degree of biosafety, it is suggested that the measles-mumps rubella vaccine in particular could be modified to obviate any possibility of triggering anti-myelin autoimmunity. PMID- 11015742 TI - The novel receptors that mediate the entry of herpes simplex viruses and animal alphaherpesviruses into cells. AB - An extended array of cell surface molecules serve as receptors for HSV entry into cells. In addition to the heparan sulphate glycosaminoglycans, which mediate the attachment of virion to cells, HSV requires an entry receptor. The repertoire of entry receptors into human cells includes molecules from three structurally unrelated molecular families. They are (i) HveA (herpesvirus entry mediator A), (ii) members of the nectin family, (iii) 3-O-sulphated heparan sulphate. The molecules have different attributes and play potentially different roles in HSV infection and spread to human tissues. All the human entry receptors interact physically with the virion envelope glycoprotein D (gD). (i) HveA is a member of the TNF-receptor family. It mediates entry of a restricted range of HSV strains. Its expression is restricted to few lineages (e.g. T-lymphocytes). (ii) The human nectin1alpha (HIgR), nectin1delta (PRR1-HveC), and the nectin2alpha (PRR2alpha HveB) and nectin2delta (PRR2delta) belong to the immunoglobulin superfamily. They are homologues of the poliovirus receptor (CD155), with which they share the overall structure of the ectodomain. The human nectin1alpha-delta are broadly expressed in cell lines of different lineages, are expressed in human tissue targets of HSV infection, serve as receptors for all HSV-1 and HSV-2 strains tested and mediate entry not only of free virions, but also cell-to-cell spread of virus. (iii) The 3-O-sulphated heparan sulphate is expressed in some selected human cell lines (e.g. endothelial and mast cells) and human tissues, and mediates entry of HSV-1, but not HSV-2. The human nectin2alpha and nectin2delta serve as receptors for a narrow range of viruses. A characteristic of the human nectin1alpha-delta is the promiscuous species non-specific receptor activity towards the animal alphaherpesviruses, pseudorabies virus (PrV) and bovine herpesvirus 1 (BHV-1). By contrast with the human nectin1delta, its murine homologue (mNectin1delta) does not bind gD at detectable level, yet it mediates entry of HSV, as well as of PrV and BHV-1. This provides the first example of a mediator of HSV entry independent of a detectable interaction with gD. PMID- 11015743 TI - Viral genome characterisation by the heteroduplex mobility and heteroduplex tracking assays. AB - The heteroduplex mobility assay (HMA) is a means of comparing two PCR amplicons or, in the variation known as the heteroduplex tracking assay (HTA), a means of estimating the quasispecies diversity of a viral genome. Heteroduplex assays have many applications including subtyping viral genomes, screening for low frequency variants in a population, scanning the relative genetic diversity across a genome and screening for recombinant clones. They can be used to detect dual infections, superinfections, contaminated blood products and laboratory contaminations. PCR amplicons of about 65% sequence similarity or greater will form heteroduplexes under appropriate conditions, and phylogenetic trees can be drawn from heteroduplex mobility data. While homoduplexes indicate more than 98% similarity between two DNA sequences, heteroduplexes indicate at least seven mismatches in a 500-bp amplicon, or a three-base pair gap in 1000-bp. Minority variants comprising 1% to 5% of the genome population can be detected and quantified by HTA. Thus far, heteroduplex assays have been described for HIV and other lentiviruses, hepatitis C and G viruses, Norwalk-like viruses, influenza, measles and poliovirus. They could be applied to a wide range of other viral species. PMID- 11015745 TI - E. De clerq, 'Novel compounds in preclinical/early clinical development for the treatment of HIV infections'. Reviews In medical virology 10(4) 2000, 255-277 AB - The original article to which this Erratum refers was published in Reviews in Medical Virology 10(4) 2000, 255-277. PMID- 11015744 TI - Molecular aspects of avian influenza (H5N1) viruses isolated from humans. AB - In 1997, 18 human infections with H5N1 influenza type A were identified in Hong Kong and six of the patients died. There were concomitant outbreaks of H5N1 infections in poultry. The gene segments of the human H5N1 viruses were derived from avian influenza A viruses and not from circulating human influenza A viruses. In 1999 two cases of human infections caused by avian H9N2 virus were also identified in Hong Kong. These events established that avian influenza viruses can infect humans without passage through an intermediate host and without acquiring gene segments from human influenza viruses. The likely origin of the H5N1 viruses has been deduced from molecular analysis of these and other viruses isolated from the region. The gene sequences of the H5N1 viruses were analysed in order to identify the molecular basis for the ability of these avian viruses to infect humans. PMID- 11015747 TI - Another reason to get the lead out. PMID- 11015746 TI - Heart-healthy eating. PMID- 11015748 TI - Ask the doctor. My neighbor has atherosclerosis in his leg arteries and he has been getting chelation therapy. He swears it has made a huge difference and thinks I should try it for my coronary artery disease. Your advice? PMID- 11015750 TI - Flu shots. PMID- 11015749 TI - Ask the doctor. Just two years ago, my total cholesterol was 175 mg/dL, and my LDL cholesterol was 128 mg/dL. This week my doctor told me that my cholesterol was 240 and my LDL was 170. I haven't gained any weight or changed my diet- what's going on? PMID- 11015752 TI - Surgery. Total knee replacement. PMID- 11015751 TI - Diet. Tea and health. PMID- 11015753 TI - Nutrition. Whole fruits and vegetables. PMID- 11015754 TI - By the way, doctor... In the last five years or so, I've occasionally had an intense pain on my left side. The doctors tell me that I have diverticulitis and should think about surgery. I hate the idea. Is it necessary? PMID- 11015755 TI - SSRIs: Prozac and company - part I. PMID- 11015756 TI - Adult personality and early experience. PMID- 11015757 TI - Estrogen not effective for Alzheimer's disease. PMID- 11015758 TI - Testing cocaine babies. PMID- 11015759 TI - What is the relationship between schizophrenia and substance abuse? PMID- 11015760 TI - Dining in, dining out: eating well at home and away. PMID- 11015761 TI - That lingering cough. Part II: specific causes. PMID- 11015762 TI - Testosterone for women? PMID- 11015763 TI - On call. My wife wants me to get more exercise. I play golf three or four times a week during the nice weather, and I think that's enough. Who is right? PMID- 11015764 TI - Combating antibiotic resistance. PMID- 11015765 TI - Breast cancer update, part I. PMID- 11015766 TI - Boning up on calcium. PMID- 11015767 TI - By the way, doctor. In the lead article about HRT use in your April issue, you didn't mention Estrace. I've taken it for several years, without any problems. But am I getting the same benefits as I would with Premarin, and are the risks similar? PMID- 11015768 TI - Complementary and alternative therapies and the question of evidence. PMID- 11015769 TI - Association of involvement in psychological self-regulation with longer survival in patients with metastatic cancer: an exploratory study. AB - There have been numerous anecdotal claims that when patients are dedicated users of a variety of psychological self-regulation strategies, including relaxation, mental imaging, cognitive restructuring and meditation, such dedication may have a life-prolonging effect. Our aim was to test this possibility more rigorously, in patients with metastatic cancer.A prospective, longitudinal, correlative study was carried out on 22 patients with varying kinds of medically incurable metastatic cancer. The intervention was one year of weekly group psychological therapy. Extensive verbal data (patients' written homework and therapists' notes) were collected over the year. The extent of each patient's involvement with psychological work was estimated following a qualitative analysis of these data. Patients were classed as showing high, moderate, or low involvement on the basis of a quantitative rating of categories defined by the analysis. These three subgroups did not differ significantly in their expected median survival duration as estimated from independent quantitative predictions by a large panel of oncologists who analyzed the patients' medical charts at time of study entry.A significant relationship was found between degree of involvement in psychological work and survival duration. Results are presented as Kaplan-Meier survival curves (Fig. 2;P = 0. 006, Log Rank test) and as a graphic display of the median survival of each of the three groups (Fig. 3). The main likely confounders (medical status, age, quality of life, and attendance at therapy) were similar across subgroups and did not change the relation between psychological work and survival duration. Limitations in the design are discussed. However, the strong effects observed support clinical observations that dedicated involvement in psychological self-regulation may prolong the life of some patients with metastatic cancer. PMID- 11015770 TI - Challenges to the biomedical model: are actions of patients almost always as important as actions of health professionals in long-term outcomes of chronic diseases? PMID- 11015771 TI - Mind-body health at 25: an assessment. AB - This is a personal assessment of the past 25 years of the mind-body health movement. It argues that, despite many advances, the movement is notably marred by its unwillingness to confront the economic and environmental determinants of health. PMID- 11015772 TI - Lessons about "The schizophrenia of the body": a comment on Anna Cassirer Appelbaum's "Observations on the disintegration of the self". PMID- 11015773 TI - Salient observations and concepts but dubious methodology. PMID- 11015774 TI - Effect of early mobilisation on grip strength, pinch strength and work of hand muscles in cases of closed diaphyseal fracture radius-ulna treated with dynamic compression plating. AB - AIMS: The purpose of the study was to objectively determine the effects of early mobilisation in terms of grip strength and work of hand muscles in cases of closed diaphyseal fracture radius - ulna treated with dynamic compression plating. SUBJECTS AND METHODS: Fifty normal subjects and Twenty-one patients, (Eleven patients treated with early active and resistive goal directed mobilisation and Ten control group) were assessed for pinch strength and grip strength on Pinch Dynamometer and Jamar Dynamometer and work of hand muscles on Ergograph. Standardised positions of the equipments and patients were maintained throughout the study. RESULTS: Results showed highly significant reduction in performance in patients treated with early mobilisation as compared to normal subjects in their first assessment (Fourth week post operatively). These patients showed significant improvement in successive assessments (sixth & eighth post operative week) on exercising in between these assessments. CONCLUSIONS: There are significant effects on grip strength and work of hand muscles in patients treated with that early active and resistive goal directed mobilisation. PMID- 11015776 TI - JAMA 100 years ago: THE RELATIONS OF SCIENTISTS TO NEWSPAPERS PMID- 11015775 TI - A piece of my mind: dare we go gently. PMID- 11015777 TI - JAMA 100 years ago: MEDICAL ORATORY PMID- 11015778 TI - Nosocomial TB control guidelines debated; will OSHA's proposed regulations prevail? Occupational Safety and Health Administration. PMID- 11015779 TI - Hypertension experts recommend new focus on the systolic reading. PMID- 11015780 TI - New guidelines for common cardiac disease. PMID- 11015782 TI - From the food and drug administration: new inhaled corticosteroid PMID- 11015783 TI - From the food and drug administration: new hearing implant approved PMID- 11015781 TI - From the Food and Drug Administration. PMID- 11015784 TI - Issues in establishing primary stroke centers. PMID- 11015785 TI - Issues in establishing primary stroke centers. PMID- 11015786 TI - Issues in establishing primary stroke centers PMID- 11015787 TI - Safety precautions to limit exposure from plague-infected patients. PMID- 11015788 TI - Safety precautions to limit exposure from plague-infected patients. PMID- 11015789 TI - Safety precautions to limit exposure from plague-infected patients PMID- 11015790 TI - The role of genotypic resistance testing in selecting therapy for HIV. PMID- 11015791 TI - The role of genotypic resistance testing in selecting therapy for HIV PMID- 11015792 TI - The continuing epidemic of obesity in the United States. PMID- 11015793 TI - Racial differences in knowledge regarding hepatitis C virus infection. PMID- 11015794 TI - Levels of environmental endotoxin and prevalence of atopic disease. PMID- 11015795 TI - Effectiveness and cost-benefit of influenza vaccination of healthy working adults: A randomized controlled trial. AB - CONTEXT: Although the cost-effectiveness and cost-benefit of influenza vaccination are well established for persons aged 65 years or older, the benefits for healthy adults younger than 65 years are less clear. OBJECTIVE: To evaluate the effectiveness and cost-benefit of influenza vaccine in preventing influenza like illness (ILI) and reducing societal costs of ILI among healthy working adults. DESIGN: Double-blind, randomized, placebo-controlled trial conducted during 2 influenza seasons. SETTING AND PARTICIPANTS: Healthy adults aged 18 to 64 years and employed full-time by a US manufacturing company (for 1997-1998 season, n = 1184; for 1998-1999 season, n = 1191). INTERVENTIONS: For each season, participants were randomly assigned to receive either trivalent inactivated influenza vaccine (n = 595 in 1997-1998 and n = 587 in 1998-1999) or sterile saline injection (placebo; n = 589 in 1997-1998 and n = 604 in 1998 1999). Participants in 1997-1998 were rerandomized if they participated in 1998 1999. MAIN OUTCOME MEASURES: Influenza-like illnesses and associated physician visits and work absenteeism reported in biweekly questionnaires by all participants, and serologically confirmed influenza illness among 23% of participants in each year (n = 275 in 1997-1998; n = 278 in 1998-1999); societal cost of ILI per vaccinated vs unvaccinated person. RESULTS: For 1997-1998 and 1998-1999, respectively, 95% (1130/1184) and 99% (1178/1191) of participants had complete follow-up, and 23% in each year had serologic testing. In 1997-1998, when the vaccine virus differed from the predominant circulating viruses, vaccine efficacy against serologically confirmed influenza illness was 50% (P =.33). In this season, vaccination did not reduce ILI, physician visits, or lost workdays; the net societal cost was $65.59 per person compared with no vaccination. In 1998 1999, the vaccine and predominant circulating viruses were well matched. Vaccine efficacy was 86% (P =.001), and vaccination reduced ILI, physician visits, and lost workdays by 34%, 42%, and 32%, respectively. However, vaccination resulted in a net societal cost of $11.17 per person compared with no vaccination. CONCLUSION: Influenza vaccination of healthy working adults younger than 65 years can reduce the rates of ILI, lost workdays, and physician visits during years when the vaccine and circulating viruses are similar, but vaccination may not provide overall economic benefits in most years. JAMA. 2000;284:1655-1663. PMID- 11015796 TI - Linkage of familial amyotrophic lateral sclerosis with frontotemporal dementia to chromosome 9q21-q22. AB - CONTEXT: Occasionally, 2 or more major neurodegenerative diseases arise simultaneously. An understanding of the genetic bases of combined disorders, such as amyotrophic lateral sclerosis (ALS) with frontotemporal dementia (FTD), will likely provide insight into mechanisms of these and related neurodegenerative diseases. OBJECTIVE: To identify loci that contain genes whose defects cause ALS. DESIGN: A genome-wide linkage analysis of 2 data sets from an ongoing study begun in the mid-1980s at 4 university research centers. SUBJECTS: An initial subset of 16 families (549 people) potentially informative for genetic analysis, in which 2 or more individuals were diagnosed as having ALS, identified from a Boston data set of 400 families and 4 families potentially informative (244 people) subsequently identified from a Chicago data set of more than 300 families to test a hypothesis based on findings from the Boston families. MAIN OUTCOME MEASURES: Linkage calculations assuming autosomal dominant inheritance with age-dependent penetrance (a parametric logarithm-of-odds [lod] score of 1.0 or greater required for further study of a potential locus); crossover analysis involving the ALS-FTD locus. RESULTS: In a set of families in which persons develop both ALS and FTD or either ALS or FTD alone, a genetic locus that is linked to ALS with FTD located between markers D9S301 and D9S167 was identified on human chromosome 9q21-q22. Families with ALS alone did not show linkage to this locus. Crossover analysis indicates this region covers approximately 17 cM. CONCLUSION: These data suggest that a defective gene located in the chromosome 9q21-q22 region may be linked to ALS with FTD. JAMA. 2000;284:1664-1669. PMID- 11015797 TI - Quality of medical care delivered to Medicare beneficiaries: A profile at state and national levels. AB - CONTEXT: Despite condition-specific and managed care-specific reports, no systematic program has been developed for monitoring the quality of medical care provided to Medicare beneficiaries. OBJECTIVE: To create a monitoring system for a range of measures of clinical performance that supports quality improvement and provides repeated, reliable estimates at the national and state levels for fee for-service (FFS) Medicare beneficiaries. DESIGN, SETTING, AND PARTICIPANTS: National study of repeated, cross-sectional observational data collected in 1997 1999 on all Medicare FFS beneficiaries or on a representative sample of beneficiaries with a particular condition. Data were collected using medical record abstraction for inpatient care, analysis of Medicare claims for some ambulatory services, and surveys for immunization rates. Separate samples were drawn for each topic for each state. MAIN OUTCOME MEASURES: Beneficiary patients' receipt of 24 process-of-care measures related to primary prevention, secondary prevention, or treatment of 6 medical conditions (acute myocardial infarction, breast cancer, diabetes mellitus, heart failure, pneumonia, and stroke) for which there is strong scientific evidence and professional consensus that the process of care either directly improves outcomes or is a necessary step in a chain of care that does so. RESULTS: Across all states for all measures, the percentage of patients receiving appropriate care in the median state ranged from a high of 95% (avoidance of sublingual nifedipine for patients with acute stroke) to a low of 11% (patients with pneumonia screened for pneumococcal immunization status before discharge). The median performance on an indicator is 69% (patients discharged with heart failure diagnosis who received angiotensin-converting enzyme inhibitors; diabetic patients having an eye examination in the last 2 years). Some states (particularly less populous states and those in the Northeast) consistently ranked high in relative performance while others (particularly more populous states and those in the Southeast) consistently ranked low. CONCLUSIONS: It is possible to assemble information on a diverse set of clinical performance measures that represent performance on the range of services in a health insurance program. These findings indicate substantial opportunities to improve the care delivered to Medicare beneficiaries and urgently invite a partnership among practitioners, hospitals, health plans, and purchasers to achieve that improvement. JAMA. 2000;284:1670-1676. PMID- 11015798 TI - Effectiveness of influenza vaccination of day care children in reducing influenza related morbidity among household contacts. AB - CONTEXT: A growing proportion of young children in the United States participate in day care, and these children are considered to be at high risk for influenza infection. Whether vaccinating day care children reduces household transmission of influenza is not known. OBJECTIVE: To evaluate the effect of vaccinating day care children on reducing influenza-related morbidity among their household contacts. DESIGN: Single-blind, randomized controlled trial conducted during the 1996-1997 influenza season. SETTING: Ten day care centers for children of US Navy personnel in San Diego, Calif. PARTICIPANTS: A total of 149 day care attendees (aged 24-60 months) and their families were randomized; 127 children and their 328 household contacts received 2 vaccine doses and were included in the analysis. INTERVENTIONS: Inactivated influenza vaccine was administered to 60 children with 162 household contacts, and hepatitis A vaccine as a control was administered to 67 age-matched children with 166 household contacts. MAIN OUTCOME MEASURES: Information regarding febrile respiratory illnesses and related morbidity for household contacts of influenza-vaccinated vs control children (subgrouped by influenza-vaccinated and unvaccinated contacts), obtained by telephone interviews with parents every 2 weeks from November 1996 through April 1997. RESULTS: Influenza-unvaccinated household contacts (n = 120) of influenza vaccinated day care children had 42% fewer febrile respiratory illnesses (P =.04) compared with unvaccinated household contacts of control children. Among school aged household contacts (aged 5-17 years), there was an 80% reduction among contacts of vaccinated children (n = 28) vs contacts of unvaccinated children (n = 31) in febrile respiratory illnesses (P =.01), as well as reductions of more than 70% in school days missed (P =.02), reported earaches (P =.02), physician visits (P =.007), physician-prescribed antibiotics (P =.02), and adults who missed work to take care of ill children (P =.04). CONCLUSIONS: These results indicate that vaccinating day care children against influenza helps reduce influenza-related morbidity among their household contacts, particularly among school-aged contacts. Future studies should be conducted in civilian populations to assess the full effect of vaccinating day care children against influenza. JAMA. 2000;284:1677-1682. PMID- 11015799 TI - Transmission of Mycobacterium tuberculosis from medical waste. AB - CONTEXT: Washington State has a relatively low incidence rate of tuberculosis (TB) infection. However, from May to September 1997, 3 cases of pulmonary TB were reported among medical waste treatment workers at 1 facility in Washington. There is no previous documentation of Mycobacterium tuberculosis transmission as a result of processing medical waste. OBJECTIVE: To identify the source(s) of these 3 TB infections. DESIGN, SETTING, AND PARTICIPANTS: Interviews of the 3 infected patient-workers and their contacts, review of patient-worker medical records and the state TB registry, and collection of all multidrug-resistant TB (MDR-TB) isolates identified after January 1, 1995, from the facility's catchment area; DNA fingerprinting of all isolates; polymerase chain reaction and automated DNA sequencing to determine genetic mutations associated with drug resistance; and occupational safety and environmental evaluations of the facility. MAIN OUTCOME MEASURES: Previous exposures of patient-workers to TB; verification of patient worker tuberculin skin test histories; identification of other cases of TB in the community and at the facility; drug susceptibility of patient-worker isolates; and potential for worker exposure to live M tuberculosis cultures. RESULTS: All 3 patient-workers were younger than 55 years, were born in the United States, and reported no known exposures to TB. We did not identify other TB cases. The 3 patient-workers' isolates had different DNA fingerprints. One of 10 MDR-TB catchment-area isolates matched an MDR-TB patient-worker isolate by DNA fingerprint pattern. DNA sequencing demonstrated the same rare mutation in these isolates. There was no evidence of personal contact between these 2 individuals. The laboratory that initially processed the matching isolate sent contaminated waste to the treatment facility. The facility accepted contaminated medical waste where it was shredded, blown, compacted, and finally deactivated. Equipment failures, insufficient employee training, and respiratory protective equipment inadequacies were identified at the facility. CONCLUSION: Processing contaminated medical waste resulted in transmission of M tuberculosis to at least 1 medical waste treatment facility worker. JAMA. 2000;284:1683-1688. PMID- 11015800 TI - Drug dependence, a chronic medical illness: implications for treatment, insurance, and outcomes evaluation. AB - The effects of drug dependence on social systems has helped shape the generally held view that drug dependence is primarily a social problem, not a health problem. In turn, medical approaches to prevention and treatment are lacking. We examined evidence that drug (including alcohol) dependence is a chronic medical illness. A literature review compared the diagnoses, heritability, etiology (genetic and environmental factors), pathophysiology, and response to treatments (adherence and relapse) of drug dependence vs type 2 diabetes mellitus, hypertension, and asthma. Genetic heritability, personal choice, and environmental factors are comparably involved in the etiology and course of all of these disorders. Drug dependence produces significant and lasting changes in brain chemistry and function. Effective medications are available for treating nicotine, alcohol, and opiate dependence but not stimulant or marijuana dependence. Medication adherence and relapse rates are similar across these illnesses. Drug dependence generally has been treated as if it were an acute illness. Review results suggest that long-term care strategies of medication management and continued monitoring produce lasting benefits. Drug dependence should be insured, treated, and evaluated like other chronic illnesses. JAMA. 2000;284:1689-1695. PMID- 11015801 TI - Current and future public health challenges. PMID- 11015802 TI - Preventing influenza in healthy adults: the evolving story. PMID- 11015803 TI - Further adventures of the tubercle bacillus. PMID- 11015804 TI - MSJAMA: No one an island: the geography of the whole patient. PMID- 11015805 TI - MSJAMA: No one an island: the geography of the whole patient PMID- 11015806 TI - MSJAMA: mind matters, money matters: the cost-effectiveness of mind/body medicine. PMID- 11015807 TI - MSJAMA: against the grain: living with the human immunodeficiency virus in rural South Dakota. PMID- 11015808 TI - MSJAMA: religion, spirituality, and medicine: application to clinical practice. PMID- 11015809 TI - MSJAMA: the problem with compliance in diabetes. PMID- 11015810 TI - Glucocorticoids and hippocampal atrophy in neuropsychiatric disorders. AB - An extensive literature stretching back decades has shown that prolonged stress or prolonged exposure to glucocorticoids-the adrenal steroids secreted during stress-can have adverse effects on the rodent hippocampus. More recent findings suggest a similar phenomenon in the human hippocampus associated with many neuropsychiatric disorders. This review examines the evidence for hippocampal atrophy in (1) Cushing syndrome, which is characterized by a pathologic oversecretion of glucocorticoids; (2) episodes of repeated and severe major depression, which is often associated with hypersecretion of glucocorticoids; and (3) posttraumatic stress disorder. Key questions that will be examined include whether the hippocampal atrophy arises from the neuropsychiatric disorder, or precedes and predisposes toward it; whether glucocorticoids really are plausible candidates for contributing to the atrophy; and what cellular mechanisms underlie the overall decreases in hippocampal volume. Explicit memory deficits have been demonstrated in Cushing syndrome, depression, and posttraumatic stress disorder; an extensive literature suggests that hippocampal atrophy of the magnitude found in these disorders can give rise to such cognitive deficits. PMID- 11015811 TI - The hippocampus in patients treated with electroconvulsive therapy: a proton magnetic resonance spectroscopic imaging study. AB - BACKGROUND: We monitored the effect of electroconvulsive therapy (ECT) on the nuclear magnetic resonance-detectable metabolites N-acetylaspartate, creatine and phosphocreatine, and choline-containing compounds in the hippocampus by means of hydrogen 1 magnetic resonance spectroscopic imaging. We hypothesized that if ECT induced memory deterioration was associated with neuronal loss in the hippocampus, the N-acetylaspartate signal would decrease after ECT and any increased membrane turnover would result in an increase in the signal from choline-containing compounds. METHODS: Seventeen patients received complete courses of ECT, during which repeated proton magnetic resonance spectroscopic imaging studies of the hippocampal region were performed. Individual changes during the course of ECT were compared with values obtained in 24 healthy control subjects and 6 patients remitted from major depression without ECT. RESULTS: No changes in the hippocampal N-acetylaspartate signals were detected after ECT. A significant mean increase of 16% of the signal from choline-containing compounds after 5 or more ECT treatments was observed. Despite the mostly unilateral ECT application (14 of 17 patients), the increase in the choline-containing compound signal was observed bilaterally. Lactate or elevated lipid signals were not detected. All patients showed clinical amelioration of depression after ECT. CONCLUSIONS: Electroconvulsive therapy is not likely to induce hippocampal atrophy or cell death, which would be reflected by a decrease in the N acetylaspartate signal. Compared with an age-matched control group, the choline containing compounds signal in patients with a major depressive episode was significantly lower than normal, before ECT and normalized during ECT. PMID- 11015812 TI - Mental disorders and the incidence of migraine headaches in a community sample: results from the Baltimore Epidemiologic Catchment area follow-up study. AB - BACKGROUND: The cross-sectional relation between migraine headaches and affective disorders has been demonstrated in studies of clinical and community populations. Few studies have investigated the prospective relation between psychiatric disorders and migraine headaches. METHODS: A prospective follow-up of the Baltimore, Md, cohort of the Epidemiologic Catchment Area Study assessed psychopathologic features in 1981 and again between 1993 and 1996. Interviews included a history of headaches at baseline and self-reported assessment of migraine headaches at follow-up. Risk estimates for incident migraine headaches by 1981 demographic variables and psychopathologic features were calculated. The cross-sectional association between prevalent migraine and lifetime psychiatric diagnoses was estimated. RESULTS: In the at-risk population of 1343, there were 118 incident cases of migraine headaches. The age- and sex-specific incident rates of migraine headaches followed the expected patterns, with younger age and female sex identified as risk factors. In cross-sectional analyses, major depression (odds ratio, 3.14; 95% confidence interval, 2.03-4. 84) and panic disorder (odds ratio, 5.09; 95% confidence interval, 2. 65-9.79) had the strongest associations, and alcohol and other substance abuse were not associated. In logistic regression models including age, sex, and psychiatric illness in 1981, only phobia was predictive of incident migraines (odds ratio, 1.70; 95% confidence interval, 1.11-2.58). Affective disorders were not predictive of incident migraine headaches. Including a history of tricyclic antidepressant use did not change the results. CONCLUSIONS: There is a strong cross-sectional relation between affective disorders and migraine headaches in this cohort. However, there is no association between antecedent affective disorders and incident migraine headaches in this population-based prospective study. PMID- 11015813 TI - Childhood sexual abuse and adult psychiatric and substance use disorders in women: an epidemiological and cotwin control analysis. AB - BACKGROUND: Women who report childhood sexual abuse (CSA) are at increased risk for developing psychiatric disorders in adulthood. What is the diagnostic specificity and cause of this association? METHODS: In a population-based sample of 1411 female adult twins, 3 levels of CSA were assessed by self-report and cotwin report: nongenital, genital, and intercourse. Interviews with twins and parents assessed family background and diagnoses of psychiatric and substance dependence disorders. Odds ratios (ORs) were calculated by logistic regression. RESULTS: By self-report, 30.4% reported any CSA and 8.4% reported intercourse. Self-reported CSA was positively associated with all disorders, the highest ORs being seen with bulimia and alcohol and other drug dependence. The ORs were modest and often nonsignificant with nongenital CSA and increased with genital CSA and especially intercourse, where most ORs exceeded 3.0. A similar pattern of findings was seen with CSA as reported by the cotwin, although many ORs were smaller. Controlling for family background factors and parental psychopathology produced a small to modest reduction in ORs. In twin pairs discordant for CSA, the exposed twin was at consistently higher risk of illness. CONCLUSIONS: Women with CSA have a substantially increased risk for developing a wide range of psychopathology. Most of this association is due to more severe forms of CSA and cannot be explained by background familial factors. Although other biases cannot be ruled out, these results are consistent with the hypothesis that CSA is causally related to an increased risk for psychiatric and substance abuse disorders. PMID- 11015814 TI - Differential carbon dioxide sensitivity in childhood anxiety disorders and nonill comparison group. AB - BACKGROUND: To examine the relationship between respiratory regulation and childhood anxiety disorders, this study considered the relationship between anxiety disorders and symptoms during carbon dioxide (CO(2)) exposure, CO(2) sensitivity in specific childhood anxiety disorders, and the relationship between symptomatic and physiological responses to CO(2). METHODS: Following procedures established in adults, 104 children (aged 9-17 years), including 25 from a previous study, underwent 5% CO(2) inhalation. The sample included 57 probands with an anxiety disorder (social phobia, generalized anxiety disorder, separation anxiety disorder, and panic disorder) and 47 nonill comparison subjects. Symptoms of anxiety were assessed before, during, and after CO(2) inhalation. RESULTS: All children tolerated the procedure well, experiencing transient or no increases in anxiety symptoms. Children with an anxiety disorder, particularly separation anxiety disorder, exhibited greater changes in somatic symptoms during inhalation of CO(2)-enriched air, relative to the comparison group. During CO(2) inhalation, symptom ratings were positively correlated with respiratory rate increases, as well as with levels of tidal volume, minute ventilation, end-tidal CO(2), and irregularity in respiratory rate during room-air breathing. CONCLUSIONS: Childhood anxiety disorders, particularly separation anxiety disorder, are associated with CO(2) hypersensitivity, as defined by symptom reports. Carbon dioxide hypersensitivity is associated with physiological changes similar to those found in panic disorder. These and other data suggest that certain childhood anxiety disorders may share pathophysiological features with adult panic disorder. PMID- 11015815 TI - Olanzapine treatment of psychotic and behavioral symptoms in patients with Alzheimer disease in nursing care facilities: a double-blind, randomized, placebo controlled trial. The HGEU Study Group. AB - BACKGROUND: Patients with Alzheimer disease (AD) commonly exhibit psychosis and behavioral disturbances that impair patient functioning, create caregiver distress, and lead to institutionalization. This study was conducted to assess the efficacy and safety of olanzapine in treating psychosis and/or agitation/aggression in patients with AD. METHODS: A multicenter, double-blind, placebo-controlled, 6-week study was conducted in 206 elderly US nursing home residents with AD who exhibited psychotic and/or behavioral symptoms. Patients were randomly assigned to placebo or a fixed dose of 5, 10, or 15 mg/d of olanzapine. The primary efficacy measure was the sum of the Agitation/Aggression, Hallucinations, and Delusions items (Core Total) of the Neuropsychiatric Inventory-Nursing Home version. RESULTS: Low-dose olanzapine (5 and 10 mg/d) produced significant improvement compared with placebo on the Core Total (-7.6 vs -3.7 [P<.001] and -6.1 vs -3. 7 [P =.006], respectively). Core Total improvement with olanzapine, 15 mg/d, was not significantly greater than placebo. The Occupational Disruptiveness score, reflecting the impact of patients' psychosis and behavioral disturbances on the caregiver, was significantly reduced in the 5 mg/d olanzapine group compared with placebo (-2.7 vs -1.5; P =.008). Somnolence was significantly more common among patients receiving olanzapine (25.0%-35.8%), and gait disturbance occurred in those receiving 5 or 15 mg/d (19.6% and 17.0%, respectively). No significant cognitive impairment, increase in extrapyramidal symptoms, or central anticholinergic effects were found at any olanzapine dose relative to placebo. CONCLUSION: Low-dose olanzapine (5 and 10 mg/d) was significantly superior to placebo and well tolerated in treating agitation/aggression and psychosis in this population of patients with AD. PMID- 11015816 TI - Mental disorders and violence in a total birth cohort: results from the Dunedin Study. AB - BACKGROUND: We report on mental disorders and violence for a birth cohort of young adults, regardless of their contact with the health or justice systems. METHODS: We studied 961 young adults who constituted 94% of a total-city birth cohort in New Zealand, April 1, 1972, through March 31, 1973. Past-year prevalence of mental disorders was measured using standardized DSM-III-R interviews. Past-year violence was measured using self-reports of criminal offending and a search of official conviction records. We also tested whether substance use before the violent offense, adolescent excessive perceptions of threat, and a juvenile history of conduct disorder accounted for the link between mental disorders and violence. RESULTS: Individuals meeting diagnostic criteria for alcohol dependence, marijuana dependence, and schizophrenia-spectrum disorder were 1.9 (95% confidence interval [CI], 1.0-3.5), 3.8 (95% CI, 2.2-6.8), and 2.5 (95% CI, 1.1-5.7) times, respectively, more likely than control subjects to be violent. Persons with at least 1 of these 3 disorders constituted one fifth of the sample, but they accounted for half of the sample's violent crimes (10% of violence risk was uniquely attributable to schizophrenia-spectrum disorder). Among alcohol-dependent individuals, violence was best explained by substance use before the offense; among marijuana-dependent individuals, by a juvenile history of conduct disorder; and among individuals with schizophrenia-spectrum disorder, by excessive perceptions of threat and a history of conduct disorder. CONCLUSIONS: In the age group committing most violent incidents, individuals with mental disorders account for a considerable amount of violence in the community. Different mental disorders are linked to violence via different core explanations, suggesting multiple-targeted prevention strategies. PMID- 11015818 TI - Contrast detection in schizophrenia. PMID- 11015817 TI - Cost-effectiveness of clozapine compared with conventional antipsychotic medication for patients in state hospitals. AB - BACKGROUND: An open-label, randomized controlled trial compared clozapine with physicians'-choice medications among long-term state hospital inpatients in Connecticut. The goal was to examine clozapine's cost-effectiveness in routine practice for people experiencing lengthy hospitalizations. METHODS: Long-stay patients with schizophrenia in a state hospital were randomly assigned to begin open-label clozapine (n = 138) or to continue receiving conventional antipsychotic medications (n = 89). We interviewed study participants every 4 months for 2 years to assess psychiatric symptoms and functional status, and we collected continuous measures of prescribed medications, service utilization, and other costs. We used both parametric and nonparametric techniques to examine changes in cost and parametric analyses to examine changes in effectiveness. We used bootstrap techniques to estimate incremental cost-effectiveness ratios and create cost-effectiveness acceptability curves. RESULTS: Both groups incurred similar costs during the 2-year study period, with a trend for clozapine to be less costly than usual care in the second study year. Clozapine was more effective than usual care on many but not all measures. With the use of effectiveness measures that favored clozapine (extrapyramidal side effects, disruptiveness), bootstrap techniques indicated that, even when a payer is unwilling to incur any additional cost for gains in effectiveness, the probability that clozapine is more cost-effective than usual care is at least 0.80. These findings were not as evident when outcomes where clozapine was not clearly superior (psychotic symptoms, weight gain) were examined. CONCLUSION: Clozapine demonstrated cost-effectiveness on some but not all measures of effectiveness when the alternative was a range of conventional antipsychotic medications. PMID- 11015819 TI - Contrast detection in schizophrenia PMID- 11015820 TI - Delayed "Choice" quantum eraser AB - We report a delayed "choice" quantum eraser experiment of the type proposed by Scully and Druhl (where the "choice" is made randomly by a photon at a beam splitter). The experimental results demonstrate the possibility of delayed determination of particlelike or wavelike behavior via quantum entanglement. The which-path or both-path information of a quantum can be marked or erased by its entangled twin even after the registration of the quantum. PMID- 11015821 TI - Condensation of "Composite Bosons" in a rotating BEC AB - We provide evidence for several novel phases in the dilute limit of rotating Bose Einstein condensates. By exact calculation of wave functions and energies for small numbers of particles, we show that the states near integer angular momentum per particle are best considered condensates of composite entities, involving vortices and atoms. We are led to this result by explicit comparison with a description purely in terms of vortices. Several parallels with the fractional quantum Hall effect emerge, including the presence of the Pfaffian state. PMID- 11015822 TI - Radiative tail of realistic rotating gravitational collapse AB - An astrophysically realistic model of wave dynamics in black-hole spacetimes must involve a nonspherical background geometry with angular momentum. We consider the evolution of gravitational (and electromagnetic) perturbations in rotating Kerr spacetimes. We show that a rotating Kerr black hole becomes "bald" slower than the corresponding spherically symmetric Schwarzschild black hole. Moreover, our results turn over the traditional belief (which has been widely accepted during the last three decades) that the late-time tail of gravitational collapse is universal. Our results are also of importance both to the study of the no-hair conjecture and the mass-inflation scenario (stability of Cauchy horizons). PMID- 11015823 TI - Cosmic rays observed by the resonant gravitational wave detector NAUTILUS AB - The passage of cosmic rays has been observed to excite mechanical vibrations in the resonant gravitational wave detector NAUTILUS operating at temperature of 100 mK. A very significant correlation (more than 10 standard deviations) is found. PMID- 11015824 TI - Constraints on the phase gamma and new physics from B --> kpi decays AB - Recent results from CLEO on B-->Kpi indicate that the phase gamma may be substantially different from that obtained from other fit to the KM matrix elements in the standard model. We show that gamma extracted using B-->Kpi,pipi is sensitive to new physics occurring at loop level. It provides a powerful method to probe new physics in electroweak penguin interactions. Using effects due to anomalous gauge couplings as an example, we show that within the allowed ranges for these couplings information about gamma obtained from B-->Kpi,pipi can be very different from the standard model prediction. PMID- 11015825 TI - Effect of large supersymmetric phases on higgs production AB - If the soft supersymmetry (SUSY) breaking masses and couplings are complex and cancellations do take place in the SUSY induced contributions to the fermionic electric dipole moments, then the CP-violating soft phases can drastically modify much of the known phenomenological pattern of the minimal supersymmetric standard model. In particular, the squark loop content of the dominant Higgs production mechanism at the Large Hadron Collider, the gluon-gluon fusion mode, could be responsible for large corrections to the known cross sections. PMID- 11015826 TI - Search for rare and forbidden eta(') decays AB - We have searched for rare and forbidden decays of the eta(') meson in hadronic events at the CLEO II detector. The search is conducted on 4.80 fb(-1) of e(+)e( ) collisions at 10.6 GeV center-of-mass energy at the Cornell Electron Storage Ring. We find no signals, and set 90% confidence level upper limits of their branching fractions: B(eta(')-->e(+)e(-)eta)<2.4x10(-3), B(eta(')-->e(+)e( )pi(0))<1. 4x10(-3), B(eta(')-->e(+)e(-)gamma)<0.9x10(-3), and B(eta(')- >e&mgr;)<4.7x10(-4). We also fit the matrix element of the eta(')-->pi(+)pi(-)eta Dalitz plot with the parametrization |M|(2) = A|1+alphay|(2), where y is a linear function of the kinetic energy of the eta, and find Re (alpha) = -0.021+/-0.025. PMID- 11015827 TI - Equilibrium distribution of heavy quarks in fokker-planck dynamics AB - We obtain an explicit generalization, within Fokker-Planck dynamics, of Einstein's relation between drag, diffusion, and the equilibrium distribution for a spatially homogeneous system, considering both the transverse and longitudinal diffusion for dimension n>1. We provide a complete characterization of the equilibrium distribution in terms of the drag and diffusion transport coefficients. We apply this analysis to charm quark dynamics in a thermal quark gluon plasma for the case of collisional equilibration. PMID- 11015828 TI - One-neutron knockout from individual single-particle states of 11Be AB - The structure of the halo nucleus 11Be has been studied using the reaction 9Be(11Be,10Be+gamma)X at 60 MeV/nucleon. The ground state structure of 11Be is determined by comparing the experimental cross sections to a calculation combining spectroscopic factors from the shell model with l-dependent single particle cross sections obtained in an eikonal model. This experiment shows the dominant 1s single-particle character of the 11Be ground state and indicates a small contribution of 0d admixture in the wave function. After correction for the approximately 22% intensity to excited levels, a clean and precise distribution of parallel momentum for knockout from the 1s halo wave function is obtained for the first time. PMID- 11015829 TI - Core-excited states in the doubly magic 68Ni and its neighbor 69Cu AB - In deep-inelastic collisions of 8 MeV/nucleon 70Zn projectiles with 198Pt, we have found an 8(+) isomer with T(1/2) = 23(1) ns at 4208 keV in 68Ni; the nug(9/2) E2 effective charge was determined to be 1. 5(1)e. In 69Cu, a 19/2(-) isomer with T(1/2) = 22(1) ns at 3691 keV was identified and its decay data were calculated quite accurately by a parameter-free shell model calculation using empirical input parameters. Proton 2p- 1h excitation, fed by another T(1/2) = 39(6) ns isomer at 3827 keV, induces large collectivity in 69Cu. PMID- 11015830 TI - Balance of mass, momentum, and energy in splintering central collisions for 40Ar up to 115 MeV /Nucleon AB - For central collisions of (17-115)A MeV 40Ar+Cu, Ag, Au, an overall balance is determined for the average mass, energy, and longitudinal momentum. Light charged particles and fragments are separated into forward-focused and isotropic components in the frame of the heaviest fragment. Energy removal by the isotropic component reaches 1-2 GeV. For such high deposition energies, statistical multifragmentation models predict much more extensive nuclear disassembly than is observed. PMID- 11015831 TI - Coster-kronig decay of the ar2s hole observed by auger-threshold photoelectron coincidence spectroscopy AB - The Coster-Kronig lines associated with Ar2s decay have been resolved within the natural linewidth of the 2s hole for the first time. This was possible by a new spectroscopic technique, relying on resonance enhanced double photoionization, Auger-threshold photoelectron coincidence spectroscopy. Contrary to standard Auger spectroscopy, this technique can filter out weak components in Auger spectra corresponding to a well-defined inner-shell state and, furthermore, can achieve a resolution no longer limited by the lifetime of the inner-shell hole. PMID- 11015832 TI - Electronic excitation and thermal effects in alkali-halide cluster anions AB - We have observed electronically excited states in alkali-halide cluster anions with one excess electron. Using photoelectron spectroscopy, we have found two narrow states in (KI)-2, (NaI)-2, and (NaCl)-2, consistent with a dipole-bound electron, while larger cluster anions exhibit a single broad excited state. In the larger systems, electronic excitation is often accompanied by vibrational excitation and thus a change in cluster temperature. Such temperature changes affect cluster structure and in some cases lead to rapid thermal isomerization. PMID- 11015833 TI - Multiple ionization and fragmentation of negatively charged fullerene ions by electron impact AB - Cross sections for the electron-impact multiple ionization and fragmentation of negatively charged fullerene ions C(-)(n) ( n = 60, 70) to C(q+)(n-m) ( q = 1,2,3 and m = 0,2,4) have been measured for electron energies up to 1 keV. In the case of pure ionization all threshold energies are about 10 eV higher than the values expected. This shift, however, is not observed for the fragment ions. The experimental data indicate that there is no strong electron-electron interaction between the incident electron and the attached electron. A novel ionization mechanism is proposed which can be expected to be valid for all negatively charged molecular or cluster ions which are able to shield the attached electron from the incident electron. PMID- 11015834 TI - Dual quantum nondemolition measurements via successive soliton collisions AB - Quantum mechanics allows quantum nondemolition (QND) variables to be measured without being changed. This requires QND variables to be initially in an eigenstate and measurement backaction noise to be directed into conjugate variables. Experimental demonstrations thus require two measurements: the first to collapse variables toward an eigenstate and the second to show results identical to the first. Here, we report results from two successive soliton collision QND measurements that optical correlation measurements show to be nearly identical. PMID- 11015835 TI - Fractal survival probability fluctuations AB - We study fluctuations of survival probability in an open quantum system classically described by a map with a mixed phase space. Our results provide the first numerical support to theoretical predictions that such fluctuations have a fractal structure, quantitatively related to the algebraic decay of the classical survival probability. PMID- 11015836 TI - Controlling hyperchaos AB - For a finite-dimensional dynamical system, whose governing equations may or may not be analytically available, we show how to stabilize an unstable orbit in a neighborhood of a "fully"unstable fixed point (i.e., a fixed point at which all eigenvalues of the Jacobian matrix have modulus greater than unity). Only one of the unstable directions is to be stabilized via time-dependent adjustments of control parameters. The parameter adjustments can be optimized. PMID- 11015837 TI - Direct measurement of the wigner delay associated with the goos-Hanchen effect AB - It is shown experimentally that the nonspecular reflection of light on an interface induces a time delay, as predicted by Wigner's scattering theory. A differential femtosecond technique is used to directly isolate this delay, associated with the Goos-Hanchen spatial shift produced by a grating near a resonant Wood anomaly. A delay of 4.4 fs is observed between TE and TM pulses, in agreement with the expected Wigner delay obtained from phase shift dispersion measurements. PMID- 11015838 TI - Eigenvectors of a raman medium AB - We show the existence of discrete sets of Raman sidebands which self-consistently establish a Raman coherence and propagate without change in amplitude and relative phase. Equivalently, there exist periodic femtosecond-time-scale, temporal pulse shapes which propagate without change in shape. PMID- 11015839 TI - Quasi-phase-matching in chiral materials AB - The second-order nonlinear optical coefficients associated with chirality differ in sign for the two mirror-image forms (enantiomers) of a chiral material. Structures comprised of alternating stacks of the enantiomers can therefore be used for quasi-phase-matched frequency conversion, as we demonstrate here by second-harmonic generation from Langmuir-Blodgett films of a helicenebisquinone. Such structures could lead to new types of frequency converters in which both the second-order nonlinear response and quasi-phase-matching arise from the chirality of a material rather than its polar order. PMID- 11015840 TI - Spatial screening solitons as particles AB - Photorefractive spatial screening solitons are treated as rays using geometrical optics. The ray picture is transformed into a classical mechanics picture, in which solitons move self-consistently as particles in a potential created by the induced change in the refractive index. The Hamiltonian equations of motion are integrated to yield trajectories that agree with the optical center-of-mass trajectories. The motion in the transverse plane is found to be not central and the orbits are not closed, preventing the spiraling of solitons. PMID- 11015841 TI - Rayleigh-Benard convection in a vertically oscillated fluid layer AB - We report on the first quantitative observations of convection in a fluid layer driven by both heating from below and vertical sinusoidal oscillation. Just above onset, convection patterns are modulated either harmonically or subharmonically to the drive frequency. Single-frequency patterns exhibit nearly solid-body rotations with harmonic and subharmonic states always rotating in opposite directions. Flows with both harmonic and subharmonic responses are found near a codimension-two point, yielding novel coexisting patterns with symmetries not found in either single-frequency states. Predictions from linear stability analysis of the onset Rayleigh and wave numbers compare well with experiment, and phase boundaries for coexisting patterns track single-frequency marginal stability curves. PMID- 11015842 TI - Domain growth in ternary fluids: A level set approach AB - We analyze phase separation in ternary systems in the asymptotic hydrodynamic regime when the volume fractions and concentrations are constant. The multiphase Navier-Stokes equations are solved using a level set method. A new projection method was developed to treat multiple junctions for systems with more than two phases. It is found that surface tension ratios can alter the growth mechanism of a minority phase in the presence of two majority phases. When the minority phase wets the interface of the majority phases the domain growth rate of all three phases is initially similar to that of a symmetric binary fluid but slows down at later times. PMID- 11015843 TI - Kinetic vortex chain solution in the drift-wave plasma regime AB - A fully nonlinear Bernstein-Greene-Kruskal stationary solution is found in the form of a quasi-three-dimensional chain of electron holes coupled to hydrodynamic vortices. This new coherent structure is enabled by the trapping and depletion of resonant particles, and the cyclotron dissipation of the singular current sheets. It is expected to play an important role in the collisionless magnetic field line reconnection in the drift-wave plasma regime, where it represents a plausible saturated state. PMID- 11015844 TI - Magnetic reconnection in toroidal eta(i) mode turbulence AB - Based on three-dimensional simulations of the Braginskii equations we show that for typical plasma-edge parameters the saturation of electromagnetic toroidal eta(i) mode turbulence is controlled by the self-generation and subsequent annihilation of radial magnetic field perturbations. This should be contrasted with the electrostatic limit, where the growth of the linear eta(i) mode is terminated by the onset of sheared flow modes driven by the radial plasma streams. The impact of the saturation amplitude on the transport level is substantial and is not in accord with simple mixing length arguments, suggesting that electromagnetic effects should generally be included in simulations of eta(i) mode turbulence. PMID- 11015845 TI - Edge thermal transport barrier In LHD discharges AB - In LHD discharges a significant enhancement of the global energy confinement has been achieved for the first time in a helical device with an edge thermal barrier, which exhibits a sharp gradient at the edge of the temperature profile. Key features associated with the barrier are quite different from those seen in tokamaks: (i) almost no change in particle (including impurity) transport, (ii) a gradual formation of the barrier, (iii) a very high ratio of the edge temperature to the average temperature, and (iv) no edge relaxation phenomenon. These features are very attractive in applying the thermal barrier to future reactor grade devices. PMID- 11015846 TI - D(A) steps and 2D islands of double layer height in the SiGe(001) system AB - The surfaces of step graded, partially relaxed Si(1-x)Ge(x)/Si(001) buffers were studied by scanning tunneling microscopy. The surface slips along <110> forming the crosshatch pattern, consisting of bunches of D(A) steps of double layer height. The D(A) steps are present in regions of large surface gradients close to the slips, as well as in planar regions between the slips. These regions are also characterized by the appearance of 2D islands of double layer height. The observations can be explained by assuming the strain due to the misfit dislocations to be locally anisotropic. Anisotropic misfit strain and efficient strain relaxation by the ( 2x8) Ge reconstruction were identified as the main factors causing the unusual step structure. PMID- 11015848 TI - Isotropic fluid phases of dipolar hard spheres AB - Monte Carlo simulations are used to calculate the equation of state and free energy of dipolar hard sphere fluids at low temperatures and densities. Evidence for the existence of isotropic-fluid-isotropic-fluid phase transitions is presented and discussed. Condensation in the dipolar hard sphere fluid is unusual in that it is not accompanied by large energy or entropy changes. An explanation of this behavior is put forward. PMID- 11015847 TI - Time-resolved X-Ray diffraction from coherent phonons during a laser-induced phase transition AB - Time-resolved x-ray diffraction with picosecond temporal resolution is used to observe scattering from impulsively generated coherent acoustic phonons in laser excited InSb crystals. The observed frequencies and damping rates are in agreement with a model based on dynamical diffraction theory coupled to analytic solutions for the laser-induced strain profile. The results are consistent with a 12 ps thermal electron-acoustic phonon coupling time together with an instantaneous component from the deformation-potential interaction. Above a critical laser fluence, we show that the first step in the transition to a disordered state is the excitation of large amplitude, coherent atomic motion. PMID- 11015849 TI - Following local adsorption sites through a surface chemical reaction: CH3SH on Cu(111) AB - Studying the interaction of CH3SH, methanethiol, with Cu(111) as a model system, we demonstrate the ability of chemical-shift normal incidence x-ray standing wave field measurements to identify the local adsorption geometries of coadsorbed reaction products at different temperatures, a technical problem of broad chemical significance. In the present case the local geometries of four distinct S-containing adsorbate species (intact CH3SH, two thiolate (CH3S-) reaction intermediates and atomic S) are determined. PMID- 11015850 TI - Self-organized patterning of an insulator-on-metal system by surface faceting and selective growth: NaCl/Cu(211) AB - We report experimental results on an insulator-on-metal system which is inherently unstable against lateral pattern formation on the nanometer scale. NaCl deposition on Cu(211) at substrate temperatures >300 K leads to faceting into (311) and (111) facets and selective NaCl growth on (311) facets only, thereby creating alternating stripes of bare Cu and NaCl-covered areas. The mesoscopic restructuring process is brought about by (1) the tendency to form (100)-terminated NaCl layers, (2) epitaxial matching between NaCl(100) and Cu(311), and (3) sufficient mobility of the Cu substrate surface. PMID- 11015851 TI - Spin-orbit coupling effect on quantum hall ferromagnets with vanishing zeeman energy AB - We present the phase diagram of a ferromagnetic nu = 2N+1 quantum Hall liquid in a narrow quantum well with vanishing single-particle Zeeman splitting, varepsilon(Z), and a pronounced spin-orbit coupling. Upon decreasing varepsilon(Z) the spin-polarization field of a liquid takes, first, the easy-axis configuration, followed by the formation of a helical state which affects the transport and NMR properties of a liquid and the form of topological defects in it. PMID- 11015853 TI - U parameter of the mott-hubbard insulator 6H-SiC(0001)-(sqrt AB - The 6H-SiC(0001)-(sqrt[3]xsqrt[3])R30 degrees surface exhibits one half-filled localized dangling-bond orbital per surface unit cell. Its electronic structure can accurately be described as a Mott-Hubbard insulator. We investigate its spectrum by a spin-polarized ab initio quasiparticle calculation. The resulting band structure shows one occupied and one empty surface band, separated by a direct band gap of 1.95 eV. Since the band gap in the spectrum of the Hubbard model is directly given by the on-site Coulomb-interaction parameter U of the dangling-bond orbital, our results allow for a reliable determination of U = 1.95 eV. PMID- 11015852 TI - Singlet and triplet exciton formation rates in conjugated polymer light-emitting diodes AB - By applying a molecular orbital perturbation approach, we calculate the formation rates for singlet and triplet molecular excitons associated with intermolecular charge-transfer processes. It is found that the interchain bond-charge correlation has a strong influence on the relative probabilities for generating singlet and triplet excitons. Most importantly, application of our approach to a model system for poly-(paraphenylenevinylene) shows that the ratio between the electroluminescence and photoluminescence quantum yields generally exceeds the 25% spin-degeneracy statistical limit. PMID- 11015854 TI - Dissipationless transport in low-density bilayer systems AB - In a bilayer electronic system the layer index may be viewed as the z component of an isospin- 1 / 2. An XY isospin-ordered ferromagnetic phase was observed in quantum Hall systems and is predicted to exist at zero magnetic field at low density. This phase is a superfluid for opposite currents in the two layers. At B = 0 the system is gapless but superfluidity is not destroyed by weak disorder. In the quantum Hall case, weak disorder generates a random gauge field which probably does not destroy superfluidity. Experimental signatures include Coulomb drag and collective mode measurements. PMID- 11015855 TI - Superconductivity from flat dispersion designed in doped mott insulators AB - Routes to enhance superconducting instability are explored for doped Mott insulators. With the help of insight for criticalities of metal-insulator transitions, geometrical design of lattice structure is proposed to control the instability. A guideline is to explicitly make flat band dispersions near the Fermi level without suppressing two-particle channels. In a one-dimensional model, numerical studies show that our prescription with finite-ranged hoppings realizes large enhancement of spin-gap and pairing dominant regions. We also propose several multiband systems, where the pairing is driven by intersite Coulomb repulsion. PMID- 11015856 TI - Influence of disorder on the local density of states in high- T(c) superconducting thin films AB - Using a low temperature scanning tunneling microscope in the spectroscopic mode, we find that the disorder in a Bi(2)Sr(2)CaCu(2)O(8+delta) thin film modifies dramatically the quasiparticle local density of states. Small, but well-defined superconducting regions, coexisting with dominating semiconducting areas, show well-pronounced gap structures, similar to those observed previously in high quality single crystals. Surprisingly, between these two regions, the detailed shape of the quasiparticle spectrum is virtually identical to the pseudogap previously observed at temperatures T>T(c), or in the vortex core, at 4.2 K. Thus, the role of the disorder in destroying the superconducting phase is comparable to that of the magnetic field or thermal fluctuations. PMID- 11015857 TI - Whispering vortices AB - Experiments indicating the excitation of whispering-gallery-type electromagnetic modes by a vortex moving in an annular Josephson junction are reported. At relativistic velocities the Josephson vortex interacts with the modes of the superconducting stripline resonator giving rise to novel resonances on the current-voltage characteristic of the junction. The experimental data are in good agreement with analysis and numerical calculations based on the two-dimensional sine-Gordon model. PMID- 11015858 TI - Metallic nonsuperconducting phase and D-wave superconductivity in Zn-substituted La1.85Sr0.15CuO4 AB - Measurements of the resistivity, magnetoresistance, and penetration depth were made on films of La1.85Sr0.15CuO4, with up to 12 at. % of Zn substituted for the Cu. The results show that the quadratic temperature dependence of the inverse square of the penetration depth, indicative of d-wave superconductivity, is not affected by doping. The suppression of superconductivity leads to a metallic nonsuperconducting phase, as expected for a pairing mechanism related to spin fluctuations. The metal-insulator transition occurs in the vicinity of k(F)l approximately 1, and appears to be disorder driven, with the carrier concentration unaffected by doping. PMID- 11015859 TI - Anomalous behavior of superconducting-normal mesoscopic structures near T(c) AB - We observe a maximum in the conductance of Al/n-GaAs junctions at temperatures 20 mK lower than the superconducting transition temperature (T(c)). This is the first observation of a peak in the conductance near the superconducting transition in superconducting-normal (S/N) junctions. To accommodate this effect we calculate the full temperature dependence of the conductance of these structures, invoking quasiclassical Green's functions in the diffusive limit. In addition to the well-known low-temperature peak at temperatures on the order of the Thouless energy, we find a maximum near T(c). This peak has the same origin as the subgap conductance observed in S/N junctions at low temperatures. PMID- 11015860 TI - Monte carlo simulation with time step quantification in terms of langevin dynamics AB - For the description of thermally activated dynamics in systems of classical magnetic moments numerical methods are desirable. We consider a simple model for isolated magnetic particles in a uniform field with an oblique angle to the easy axis of the particles. For this model, a comparison of the Monte Carlo method with Langevin dynamics yields new insight to the interpretation of the Monte Carlo process, leading to the implementation of a new algorithm where the Monte Carlo step is time quantified. The numeric results for the characteristic time of the magnetization reversal are in excellent agreement with asymptotic solutions for the Neel-Brown model. PMID- 11015861 TI - Influence of dipolar interaction on magnetic properties of ultrafine ferromagnetic particles AB - We use Monte Carlo simulations to study the influence of dipolar interaction and polydispersity on the magnetic properties of single-domain ultrafine ferromagnetic particles. From the zero field cooling (ZFC)/field cooling (FC) simulations we observe that the blocking temperature T(B) clearly increases with increasing strength of interaction, but it is almost not effected by a broadening of the distribution of particle sizes. While the dependence of the ZFC/FC curves on interaction and cooling rate are reminiscent of a spin glass transition at T(B), the relaxational behavior of the magnetic moments below T(B) is not in accordance with the picture of cooperative freezing. PMID- 11015862 TI - High proton polarization by microwave-induced optical nuclear polarization at 77 K AB - Protons in naphthalene and p-terphenyl doped with pentacene have been polarized up to 32% and 18%, respectively, at liquid nitrogen temperature in a magnetic field of 0.3 T by means of microwave-induced optical nuclear polarization. The polarization was measured by nuclear magnetic resonance as well as by the neutron transmission method. The relaxation time of the proton polarization at 0.0007 T and 77 K was found to be almost 3 h and the polarization enhancement reached a record value of 8x10(4). The usefulness of the method in many areas of physics and chemistry is discussed. PMID- 11015864 TI - Coherent acoustic phonon oscillations in semiconductor multiple quantum wells with piezoelectric fields AB - Large coherent acoustic phonon oscillations were demonstrated using InGaN/GaN multiple quantum wells with piezoelectric fields. With UV femtosecond pulse excitation, photogenerated carriers screened the piezoelectric field and initiated the displacive coherent phonon oscillations. The specific phonon frequency was selected by the coupling between the periodic carrier distribution and the corresponding acoustic phonon mode. The induced acoustic phonon oscillation resulted in piezoelectric field modulation and then caused absorption variation through the quantum confined Franz-Keldysh effect. The wave vector uncertainty due to the finite sample width was found to determine the observed dephasing time. PMID- 11015863 TI - Intrinsic ferroelectric coercive field AB - The Landau-Ginzburg theory of ferroelectricity predicts the intrinsic coercive field for polarization reversal, but the observed extrinsic coercive field is always much smaller as a result of nucleation, dynamic processes not covered by the static theory. We have realized the intrinsic coercive field for the first time, in two-dimensional Langmuir-Blodgett polymer films as thin as 1 nm. The measured coercive field is in good agreement with the theoretical intrinsic value, exhibits the expected dependence on temperature, and does not depend on thickness below 15 nm. PMID- 11015865 TI - Optical signatures of energy-level statistics in a disordered quantum system AB - Time-resolved measurements of the resonant Rayleigh scattering from quantum well excitons are shown to provide information on the energy-level statistics of the localized exciton states. The signal transients are reproduced by a microscopic quantum model of the exciton two-dimensional motion in presence of spatially correlated disorder. This model allows quantitative determination of the average energy separation between the localized states. Here this quantity turns out to be only a few times smaller than the average disorder amplitude, proving that spatial correlation and quantum mechanics are equally important in the description of the exciton localization process. PMID- 11015866 TI - Bound states in optical absorption of semiconductor quantum wells containing a two-dimensional electron Gas AB - The dependence of the optical absorption spectrum of a semiconductor quantum well on two-dimensional electron concentration n(e) is studied using CdTe samples. The trion peak (X-) seen at low n(e) evolves smoothly into the Fermi edge singularity at high n(e). The exciton peak (X) moves off to high energy, weakens, and disappears. The X,X- splitting is linear in n(e) and closely equal to the Fermi energy plus the trion binding energy. For Cd0.998Mn0.002Te quantum wells in a magnetic field, the X,X- splitting reflects unequal Fermi energies for M = +/-1/2 electrons. The data are explained by Hawrylak's theory of the many-body optical response including spin effects. PMID- 11015867 TI - Error thresholds for quasispecies on dynamic fitness landscapes. AB - In this paper we investigate error thresholds on dynamic fitness landscapes. We show that there exists both a lower and an upper threshold, representing limits to the copying fidelity of simple replicators. The lower bound can be expressed as a correction term to the error threshold present on a static landscape. The upper error threshold is a new limit that only exists on dynamic fitness landscapes. We also show that for long genomes and/or highly dynamic fitness landscapes there exists a lower bound on the selection pressure required for the effective selection of genomes with superior fitness independent of mutation rates, i.e. there are distinct nontrivial limits to evolutionary parameters in dynamic environments. PMID- 11015869 TI - Comment on "Inelastic collapse of a randomly forced particle" PMID- 11015868 TI - Comment on "DNA molecular cousin of Schrodinger's cat: a curious example of quantum measurement". PMID- 11015870 TI - Cornell et al. reply: PMID- 11015871 TI - Comment on "Delocalization in the 1D anderson model with long-range correlated disorder" PMID- 11015872 TI - De moura and lyra reply: PMID- 11015873 TI - Backward-to-forward jump rates on a tilted periodic substrate AB - Driven diffusion of a Brownian particle along a one-dimensional lattice is investigated numerically on decreasing its damping constant. The notions of multiple jumps, jump reversal, and backward-to-forward rates are discussed in detail. In particular, we conclude that in the underdamped limit backward jumps are suppressed relative to forward jumps more effectively than previously assumed. The dependence of such a drive-controlled mechanism on the damping constant and the temperature is interpreted analytically. PMID- 11015874 TI - Rotators with long-range interactions: connection with the mean-field approximation AB - We analyze the equilibrium properties of a chain of ferromagnetically coupled rotators which interact through a force that decays as r(-alpha) where r is the interparticle distance and alpha>/=0. By integrating the equations of motion we obtain the microcanonical time averages of both the magnetization and the kinetic energy. We detect three different regimes depending on whether alpha belongs to the intervals [0,1), (1,2), or (2,infinity). For 0bW with W-->lnu to measure the helicity of the W bosons in the top quark rest frame. Our measurement uses a t&tmacr; sample isolated in 106+/-4 pb(-1) of data collected in p&pmacr; collisions at sqrt[s] = 1.8 TeV with the CDF detector at the Fermilab Tevatron. Assuming a standard V-A weak decay, we find that the fraction of W's with zero helicity in the top rest frame is F0 = 0.91+/-0. 37(stat)+/-0.13(syst), consistent with the standard model prediction of F0 = 0.70 for a top mass of 175 GeV/c(2). PMID- 11015877 TI - Measurement of the W boson mass using electrons at large rapidities AB - We report a measurement of the W boson mass based on an integrated luminosity of 82 pb(-1) from p&pmacr; collisions at sqrt[s] = 1.8 TeV recorded in 1994-1995 by the D0 detector at the Fermilab Tevatron. We identify W bosons by their decays to enu, where the electron is detected in the forward calorimeters. We extract the mass by fitting the transverse mass and the electron and neutrino transverse momentum spectra of 11 089 W boson candidates. We measure M(W) = 80.691+/-0.227 GeV. By combining this measurement with our previously published central calorimeter results from data taken in 1992-1993 and 1994-1995, we obtain M(W) = 80.482+/-0.091 GeV. PMID- 11015878 TI - Higgs-mediated B0 --> &mgr;(+)&mgr;(-) in minimal supersymmetry AB - We demonstrate a new source for flavor-changing neutral currents within the minimal supersymmetric standard model. At moderate to large tanbeta, it is no longer possible to diagonalize the masses of the quarks in the same basis as their Yukawa couplings. This generates flavor-violating couplings of the form &bmacr;(R)d(L)straight phi and &bmacr;(R)s(L)straight phi where straight phi is any of the three neutral, physical Higgs bosons. These new couplings lead to rare processes in the B system such as B0-->&mgr;(+)&mgr;(-) and B0-&Bmacr;(0) mixing. We show that the latter are anomalously suppressed, while the former is in the experimentally interesting range, with an observable signal possible at Run II of the Tevatron if m(A) less, similar400-700 GeV. PMID- 11015879 TI - Observation of diffractive b-quark production at the fermilab tevatron AB - We report a measurement of the fraction of b quarks produced diffractively in &pmacr;p collisions at sqrt[s] = 1.8 TeV. Diffraction is identified by the absence of particles in a forward pseudorapidity region. From events with an electron of transverse momentum 9.5varepsilonsigma(u) channel as it was believed earlier on the basis of single particle calculations, but in both 1sigma(g)-->varepsilonsigma(u) and 1sigma(u)- >varepsilonsigma(g) channels. As a confirmation of this phenomenon we show that the experimental angular distributions of photoelectrons ejected from fixed-in space N2 molecules can be reproduced theoretically only after taking into account many-electron correlations. PMID- 11015884 TI - Collective effects in the self-interference of a single photon emitted by two atoms AB - Collective effects in the spontaneous emission pattern of two identical two-level atoms a fixed distance apart and sharing initially a single excitation are investigated. It is shown that the interference can take place even when it is known for certain which atom is excited initially. This interference is due solely to the atomic coherence established through multiple photon absorptions and reemissions and will disappear if it is ignored. The interference patterns with and without collective effects are compared for symmetric and antisymmetric initial states. The dark center from an antisymmetric state is shown both analytically and numerically. PMID- 11015885 TI - Chaotic transport and current reversal in deterministic ratchets AB - We address the problem of the classical deterministic dynamics of a particle in a periodic asymmetric potential of the ratchet type. We take into account the inertial term in order to understand the role of the chaotic dynamics in the transport properties. By a comparison between the bifurcation diagram and the current, we identify the origin of the current reversal as a bifurcation from a chaotic to a periodic regime. Close to this bifurcation, we observed trajectories revealing intermittent chaos and anomalous deterministic diffusion. PMID- 11015886 TI - Quantization of a billiard model for interacting particles AB - We consider a billiard model of a self-bound, interacting three-body system in two spatial dimensions. Numerical studies show that the classical dynamics is chaotic. The corresponding quantum system displays spectral fluctuations that exhibit small deviations from random matrix theory predictions. These can be understood in terms of a new type of scarring caused by a one-parameter family of orbits inside the collinear manifold. PMID- 11015887 TI - Pulsed gradient spin echo nuclear magnetic resonance imaging of diffusion in granular flow AB - We derive the formalism to obtain spatial distributions of collisional correlation times for macroscopic particles undergoing granular flow from pulsed gradient spin echo nuclear magnetic resonance diffusion data. This is demonstrated with an example of axial motion in the shear flow regime of a 3D granular flow in a horizontal rotating cylinder at one rotation rate. PMID- 11015888 TI - Parametric X rays observed under bragg condition: boost of intensity by a factor of Two AB - Parametric x rays (PXR) produced by bombarding silicon and diamond crystals with electrons of 30 to 87 MeV were detected at 180 degrees relative to the direction of the electron beam. It was found that the dependence of the intensity on the orientation of the crystal agrees with the predictions of the kinematical theory of PXR. The absolute intensity is twice as large as predicted. These findings can be explained considering dynamical effects that govern the x-ray crystal interaction. Additionally, x rays caused by self-diffracted transition radiation have been observed. PMID- 11015889 TI - Detailed measurements of a diffusive supersonic wave in a radiatively heated foam AB - We have made the first detailed measurements of a diffusive supersonic radiation wave in the laboratory. A 10 mg/cm(3) SiO2 foam is radiatively heated by the x ray flux from a laser-irradiated hohlraum. The resulting radiation wave propagates axially through the optically thick foam and is measured via time resolved x-ray imaging as it breaks out the far end. The data show that the radiation wave breaks out at the center prior to breaking out at the edges, indicating a significant curvature in the radiation front. This curvature is primarily due to energy loss into the walls surrounding the foam. PMID- 11015890 TI - Resonant instability of laser filaments in a plasma AB - The stability of nonlinear laser light filaments in a homogeneous isothermal plasma with respect to coupled electromagnetic and density perturbations is examined. In addition to the previously known modulational instability of a trapped electromagnetic mode, a new fast growing resonant instability is found. It corresponds to the growth of an excited eigenmode in the waveguide formed by the filament density depletion, the associated density response being supersonic and transversally localized. The evolution of the instability is illustrated by numerical simulations in two and three spatial dimensions. PMID- 11015891 TI - Experimental confirmation of impurity convection driven by the ion-temperature gradient in toroidal plasmas AB - Strong, outward convection of low- Z impurity ions has been observed in DIII-D plasmas which have reduced anomalous transport, a weak density gradient, and a strong ion-temperature gradient. Comparing the measurements with theoretical predictions of collisional (neoclassical) transport indicates that the observed outward convection results from an effect known as "temperature screening." Taking into account the non-negligible effect of anomalous transport, quantitative agreement is found between the measured transport properties and the predicted values, including the strong Z dependence. PMID- 11015892 TI - Criterion for dominance of directional over size fluctuations in destroying order AB - For systems that exhibit a second-order phase transition with a spontaneously broken continuous O(N) symmetry at low temperatures, we give a criterion for judging at which temperature T(K) long-range directional fluctuations of the order field destroy the order when approaching the critical temperature from below. The temperature T(K) lies always significantly below the famous Ginzburg temperature T(G) at which size fluctuations of finite range become important. PMID- 11015893 TI - Mechanical flocculation in flowing fiber suspensions AB - Non-Brownian fibers commonly flocculate in flowing suspensions. A particle level simulation technique modeling fibers as chains of rods connected by hinges is developed to probe flocculation. Simulations show that flocculation can be induced solely by interfiber friction-attractive forces between fibers are not necessary. Simulated mechanical floc characteristics are consistent with experimental observations. In contrast, simulations of flocs formed by attractive forces behave qualitatively differently. PMID- 11015894 TI - Adsorptionlike collapse of diblock copolymers AB - A linear copolymer made of two reciprocally attracting N-monomer blocks collapses to a compact phase through a novel transition, whose exponents are determined with extensive Monte Carlo simulations in two and three dimensions. In the former case, an identification with the statistical geometry of suitable percolation paths allows one to predict that the number of contacts between the blocks grows like N9/16. In the compact phase the blocks are mixed and, in two dimensions, also zipped, in such a way to form a spiral, double chain structure. PMID- 11015895 TI - Self-similar chain conformations in polymer gels AB - We use molecular dynamics simulations to study the swelling of randomly end-cross linked polymer networks in good solvent conditions. We find that the equilibrium degree of swelling saturates at Q(eq) approximately N(3/5)(e) for mean strand lengths &Nmacr;(s) exceeding the melt entanglement length N(e). The internal structure of the network strands in the swollen state is characterized by a new exponent nu = 0.72+/-0.02. Our findings can be rationalized by a Flory argument for a self-similar structure of mutually interpenetrating network strands, agree partially with the classical Flory-Rehner theory, and are in contradiction to de Gennes' c(*)-theorem. PMID- 11015896 TI - Oxygen-related intrinsic defects in glassy SiO2: interstitial ozone molecules AB - Interstitial O3 molecules in 7.9 eV photon-irradiated silica are identified. Their optical absorption band at 4.8 eV nearly coincides with the 4.8 eV band of nonbridging oxygen hole centers. The O3-related band is distinguished by a smaller halfwidth (0.84 vs 1. 05 eV), by susceptibility to ultraviolet bleaching, by lack of correlation to the 1.9 eV luminescence band, and by rise of a singlet O2 luminescence band at 0.974 eV during photobleaching. This identification solves a long controversy on the nature of optical bands in silica and gives a tool for studying the mobility of atomic oxygen in SiO2. PMID- 11015897 TI - Thermodynamical liquid-glass transition in a lennard-jones binary mixture AB - We use results derived in the framework of the replica approach to study the liquid-glass thermodynamic transition. The main results are derived without using replicas and applied to the study of the Lennard-Jones binary mixture introduced by Kob and Andersen. We find that there is a phase transition due to the entropy crisis. We compute both analytically and numerically the value of the phase transition point T(K) and the specific heat in the low temperature phase. PMID- 11015898 TI - Scaling and universality of ac conduction in disordered solids AB - Recent scaling results for the ac conductivity of ionic glasses by Roling et al. [Phys. Rev. Lett. 78, 2160 (1997)] and Sidebottom [Phys. Rev. Lett. 82, 3653 (1999)] are discussed. We prove that Sidebottom's version of scaling is completely general. A new approximation to the universal ac conductivity arising in the extreme disorder limit of the symmetric hopping model, the "diffusion cluster approximation," is presented and compared to computer simulations and experiments. PMID- 11015899 TI - Bose-einstein condensation and spatial correlations in 4He AB - A new Monte Carlo method for calculating ground-state properties of liquid 4He is described. It is shown that Bose-Einstein condensation (BEC) implies delocalization of the wave function. It is shown that there is no general connection between the static structure factor and BEC. It is suggested that the observed connection in liquid 4He is due to the creation of spaces in the liquid structure, which are required so that the wave function can delocalize, in the presence of the hard-core interactions. It is shown that this suggestion is quantitatively consistent with observations on liquid 4He. PMID- 11015900 TI - Stress domains in Si(111)/a-Si3N4 nanopixel: ten-million-atom molecular dynamics simulations on parallel computers AB - Parallel molecular dynamics simulations are performed to determine atomic-level stresses in Si(111)/Si(3)N4(0001) and Si(111)/a-Si3N4 nanopixels. Compared to the crystalline case, the stresses in amorphous Si3N4 are highly inhomogeneous in the plane of the interface. In silicon below the interface, for a 25 nm square mesa stress domains with triangular symmetry are observed, whereas for a rectangular, 54 nmx33 nm, mesa tensile stress domains ( approximately 300 A) are separated by Y-shaped compressive domain wall. Maximum stresses in the domains and domain walls are -2 GPa and +2 GPa, respectively. PMID- 11015901 TI - Dislocation emission at the Silicon/Silicon nitride interface: A million atom molecular dynamics simulation on parallel computers AB - Mechanical behavior of the Si(111)/Si(3)N4(0001) interface is studied using million atom molecular dynamics simulations. At a critical value of applied strain parallel to the interface, a crack forms on the silicon nitride surface and moves toward the interface. The crack does not propagate into the silicon substrate; instead, dislocations are emitted when the crack reaches the interface. The dislocation loop propagates in the (1; 1;1) plane of the silicon substrate with a speed of 500 (+/-100) m/s. Time evolution of the dislocation emission and nature of defects is studied. PMID- 11015902 TI - Facet ridge end points in crystal shapes AB - We study equilibrium crystal shapes (ECS) near facet ridge end points (FRE) by means of a numerical study of a body-centered solid-on-solid model on a square lattice with an enhanced uniaxial interaction range. This tests the stability of the so-called stochastic FRE point where the model maps exactly onto one dimensional Kardar-Parisi-Zhang-type growth and where the local ECS is simple. We find that the generic shapes are more complex. They contain first-order faceted to rough boundaries terminating in Pokrovsky-Talapov-type end points, and first order ridges inside the rounded part of the ECS where two rough surface orientations coexist. PMID- 11015903 TI - Direct observation of subcritical fluctuations during the formation of strained semiconductor islands AB - We have directly imaged subcritical fluctuations during the nucleation phase of three-dimensional islands in strained layer epitaxy. The fluctuations are defect mediated and are found to be large even at low growth temperatures. We attribute the existence of large fluctuations to the time dependence of the supersaturation. This indicates classical nucleation concepts are relevant, even at low growth temperatures. PMID- 11015904 TI - Nonuniform composition profile in In0.5Ga0.5As alloy quantum dots AB - We use cross-sectional scanning tunneling microscopy to examine the shape and composition distribution of In0.5Ga0.5As quantum dots (QDs) formed by capping heteroepitaxial islands. The QDs have a truncated pyramid shape. The composition appears highly nonuniform, with an In-rich core having an inverted-triangle shape. Thus the electronic properties will be drastically altered, relative to the uniform composition generally assumed in device modeling. Theoretical analysis of the QD growth suggests a simple explanation for the unexpected shape of the In-rich core. PMID- 11015905 TI - Collapse dynamics of liquid bridges investigated by time-varying magnetic levitation AB - Using a novel technique that facilitates temporal control over the total body force on a liquid, an unexpected scaling relationship was discovered for the collapse time of a liquid bridge. A paramagnetic liquid was suspended between the tips of two collinear rods in a strong magnetic field gradient that was adjusted to compensate gravity. A sudden change of the magnet current, corresponding to a change of Bond number, resulted in a deformation and ultimate collapse of the liquid bridge. The collapse time was found to be independent of the bridge length when other parameters were held constant. PMID- 11015906 TI - Gap-anisotropic model for the narrow-Gap kondo insulators AB - A theory is presented which accounts for the dynamical generation of a hybridization gap with nodes in the Kondo insulating materials CeNiSn and CeRhSb. We show that Hund's interactions acting on virtual 4f(2) configurations of the cerium ion can act to dynamically select the shape of the cerium ion by generating a Weiss field which couples to the shape of the ion. In low symmetry crystals where the external crystal fields are negligible, this process selects a nodal Kondo semimetal state as the lowest energy configuration. PMID- 11015907 TI - Zero-point fluctuations and the quenching of the persistent current in normal metal rings AB - The ground state of a phase-coherent mesoscopic system is sensitive to its environment. We investigate the persistent current of a ring with a quantum dot which is capacitively coupled to an external circuit with a dissipative impedance. At zero temperature, zero-point quantum fluctuations lead to a strong suppression of the persistent current with decreasing external impedance. We emphasize the role of displacement currents in the dynamical fluctuations of the persistent current and show that with decreasing external impedance the fluctuations exceed the average persistent current. PMID- 11015908 TI - Composite fermion hofstadter problem: partially polarized density wave states in the nu = 2/5 fractional quantum hall effect AB - It is well known that the nu = 2/5 state is unpolarized at zero Zeeman energy, while it is fully polarized at large Zeeman energies. A novel state with a charge/spin density wave order for composite fermions is proposed to exist at intermediate values of the Zeeman coupling for nu = 2/5. This state has half the maximum possible polarization, and can be extended to other incompressible fractions. A Hartree-Fock calculation based on the new approach for all fractional quantum Hall states developed by R. Shankar and the author is used to demonstrate the stability of this state to single-particle excitations and to compute gaps. A very recent experiment shows direct evidence for this state. PMID- 11015909 TI - NMR determination of 2D electron spin polarization at nu = 1/2 AB - Using a "standard" NMR spin-echo technique we determined the spin polarization P of two-dimensional electrons, confined to GaAs quantum wells, from the hyperfine shift of Ga nuclei located in the wells. Concentrating on the temperature ( 0.05 less, similarT less, similar10 K) and magnetic field ( 7 less, similarB less, similar17 T) dependencies of P at Landau level filling factor nu = 1/2, we find that the results are described well by a simple model of noninteracting composite fermions, although some inconsistencies remain when the two-dimensional electron system is tilted in the magnetic field. PMID- 11015910 TI - Carbon-atom wires: charge-transfer doping, voltage drop, and the effect of distortions AB - We present first-principles calculations on electrical conduction through carbon atomic wires. The changes in charge distribution induced by a large bias exhibit the primary involvement of the wire's pi states. A significant fraction ( approximately 40%) of the voltage drops across the atomic wire itself. At zero bias, there is a large transfer of charge from the electrodes to the wire, effectively providing doping without introducing scattering centers. This transfer leads, however, to potential barriers at the wire-electrode junctions. Bending the wire reduces its conductance. PMID- 11015911 TI - Negative differential conductivity in carbon nanotubes AB - A theoretical model and computations of the I-V characteristics of long carbon nanotubes in a strong axial dc field at room temperature is presented. Negative differential conductivity is predicted. It is shown that |dI/dV| for metal carbon nanotubes in the region of the negative differential conductivity significantly exceeds corresponding values for semiconducting ones. The predicted effect would enable the design of wave-generating nanotube-based diodes for submillimeter and infrared ranges. PMID- 11015912 TI - Spin dynamics and magnetic correlation length in two-dimensional quantum heisenberg antiferromagnets AB - The correlated spin dynamics and temperature dependence of the correlation length xi(T) in two-dimensional quantum (S = 1/2) Heisenberg antiferromagnets (2DQHAF) on a square lattice are discussed in light of experimental results of proton spin lattice relaxation in copper formiate tetradeuterate. In this compound the exchange constant is much smaller than the one in recently studied 2DQHAF, such as La2CuO4 and Sr2CuO2Cl2. Thus the spin dynamics can be probed in detail over a wider temperature range. The NMR relaxation rates turn out to be in excellent agreement with a theoretical mode-coupling calculation. The deduced temperature behavior of xi(T) is in agreement with high-temperature expansions, quantum Monte Carlo simulations, and the pure quantum self-consistent harmonic approximation. Contrary to the predictions of the theories based on the nonlinear sigma model, no evidence of crossover between different quantum regimes is observed. PMID- 11015913 TI - Double-spin-flip resonance of rhodium nuclei at positive and negative spin temperatures AB - Sensitive SQUID-NMR measurements were used to study the mutual interactions in the highly polarized nuclear-spin system of rhodium metal. The dipolar coupling gives rise to a weak double-spin-flip resonance. The observed frequency shifts allow deducing separately the dipolarlike contribution and the isotropic exchange term. For the first time, such measurements were extended to negative absolute temperatures as well. We find an effective dipolar moment 0. 10&mgr;(N) of which about 15% is attributed to a conduction electron mediated pseudodipolar interaction. The isotropic exchange is described by R = -0.9+/-0.1. PMID- 11015914 TI - Atom-selective bond breaking in a chemisorbed homonuclear molecule induced by core excitation: N2/Ru(001) AB - We observe photochemical selectivity for N 1s to pi(*) excitations of chemisorbed N2. By narrow bandwidth synchrotron radiation we selectively excite one of the two atoms of the molecule. Photon stimulated desorption of neutral N atoms predominates for excitations of the N atom close to the surface, whereas excitation of the outer atom ejects predominantly N02 and small amounts of N+, demonstrating the predominant breaking of the inner or outer bond, respectively, of the N2 adsorbate. Analysis on the basis of previously obtained decay electron spectra after atom-selective excitation can explain the mechanism of localized bond breaking. PMID- 11015915 TI - Angular studies of potential electron emission in the interaction of slow ions with Al surfaces AB - We report energy distributions of electrons emitted from Al surfaces under impact by 1 keV Ar+ and 1-5 keV Ne+ ions. The variation of the energy distributions with the angle of incidence is different for both ions and provides information on the mechanism responsible for electron emission. For Ar+ electron emission results mainly from Auger neutralization, while for Ne+ an important emission mechanism is the decay of plasmon excitations. We find a transition between surface and bulk plasmon excitations as the energy of the ion is increased. PMID- 11015916 TI - Presence of many stable nonhomogeneous states in an inertial car-following model AB - We present a single lane car- following model of traffic flow which is inertial and free of collisions. It demonstrates observed features of traffic flow such as existence of three regimes: free, nonhomogeneous congested (NHC) or synchronized, and homogeneous congested (HC) or jammed flow; bistability of free and NHC flow states in a range of densities, hysteresis in transitions between these states; jumps in the density-flux plane in the NHC regime; gradual spatial transition from synchronized to free flow; long survival time of jams in the HC regime. The model predicts that in the NHC regime there exist many stable states with different wavelengths, and noise can cause transitions between them. PMID- 11015917 TI - Mean-field HP model, designability and alpha-helices in protein structures. AB - Analysis of the geometric properties of a mean-field HP model on a square lattice for protein structure shows that structures with a large number of switchbacks between surface and core sites are chosen favorably by peptides as unique ground states. Global comparison of model (binary) peptide sequences with concatenated (binary) protein sequences listed in the Protein Data Bank and the Dali Domain Dictionary indicates that the highest correlation occurs between model peptides choosing the favored structures and those portions of protein sequences containing alpha helices. PMID- 11015918 TI - Thermal unbinding of highly oriented phospholipid membranes. AB - We present a temperature dependent x-ray reflectivity study of highly oriented, fully hydrated multilamellar phospholipid membranes. Both the specular and diffuse (nonspecular) x-ray reflectivity were measured for dimyristoyl-sn-glycero phosphocholine (DMPC) and oleoyl-palmitoyl-sn-glycero-phosphocholine (POPC) on silicon substrates in excess water. In this configuration the repeat distance as well as the fluctuation spectra can be determined as a function of temperature. Both model systems studied exhibit a discontinuous unbinding transition from a substrate bound, multilamellar state to a state of freely dispersed bilayers in water. In the unbound phase a single membrane remains on the substrate. PMID- 11015919 TI - Comment on "Universal relation between the kolmogorov-sinai entropy and the thermodynamical entropy in simple liquids" PMID- 11015920 TI - Dzugutov replies: PMID- 11015921 TI - Biot slow wave in a magnetorheological slurry? PMID- 11015922 TI - Nahmad-molinari et al. reply: PMID- 11015923 TI - Modulated optical lattice as an atomic fabry-perot interferometer AB - We propose to engineer the atomic band structure in optical lattices in order to design a Fabry-Perot interferometer with large mode spacing and strong nonlinear coupling to be employed in atom optics. The use of an optical lattice allows for a significant reduction of the atomic effective mass, while the slow modulation of its parameters spatially confines the matter waves on a length scale of a few dozen optical wavelengths. As a consequence, the mode spacing in such a cavity would be as high as one-tenth of the recoil energy, allowing for a very efficient filter action, while the nonlinear coupling due to interatomic interactions could lead to bistability and limiting effects in the transmission of the atomic beam. PMID- 11015924 TI - Enhancement of stochastic resonance in distributed systems due to a selective coupling AB - Recent massive numerical simulations have shown that the response of a "stochastic resonator" is enhanced as a consequence of spatial coupling. Similar results have been analytically obtained in a reaction-diffusion model, using nonequilibrium potential techniques. We now consider a field-dependent diffusivity and show that the selectivity of the coupling is more efficient for achieving stochastic-resonance enhancement than its overall value in the constant diffusivity case. PMID- 11015925 TI - Observation of CP violation in K(L)-->pi(+)pi(-)e(+)e(-) decays AB - We report the first observation of a manifestly CP violating effect in the K(L)- >pi(+)pi(-)e(+)e(-) decay mode. A large asymmetry was observed in the distribution of these decays in the CP-odd and T-odd angle straight phi between the decay planes of the e(+)e(-) and pi(+)pi(-) pairs in the K(L) center of mass system. After acceptance corrections, the overall asymmetry is found to be [13.6+/-2. 5(stat)+/-1.2(syst)]%. This is the largest CP-violating effect yet observed when integrating over the entire phase space of a mode and the first such effect observed in an angular variable. PMID- 11015926 TI - Shell correction energy for bubble nuclei AB - The positioning of a bubble inside a many fermion system does not affect the volume, surface, or curvature terms in the liquid drop expansion of the total energy. Besides possible Coulomb effects, the only other contribution to the ground state energy of such a system arises from shell effects. We show that the potential energy surface is a rather shallow function of the displacement of the bubble from the center and in most cases the preferential position of a bubble is off-center. Systems with bubbles are expected to have bands of extremely low lying collective states, corresponding to various bubble displacements. PMID- 11015928 TI - Band structure from random interactions AB - The anharmonic vibrator and rotor regions in nuclei are investigated in the framework of the interacting boson model using an ensemble of random one- and two body interactions. We find a predominance of L(P) = 0(+) ground states, as well as strong evidence for the occurrence of both vibrational and rotational band structures. This remarkable result suggests that such band structures represent a far more general (robust) property of the collective model space than is generally thought. PMID- 11015927 TI - Stringent tests of shell model calculations in fp shell nuclei (46, 48)Ti and (50,52)Cr from measurements of g factors and B(E2) values AB - Measurements of magnetic moments and lifetimes of 2(+)(1) and 4(+)(1) states of (46,48)Ti and (50,52)Cr were performed with high accuracy via projectile Coulomb excitation and the technique of transient magnetic fields. The high quality of the data allows for the first time to establish stringent constraints on large scale shell model calculations. Whereas the global behavior of the data is well explained by full fp shell model calculations, distinct deviations in the g factors and B(E2) values of (46,48)Ti from theoretical predictions can be attributed to excitations of the 40Ca core. This suggestion is supported by recent Monte Carlo calculations which provide evidence that 48Ca is a better inert core. PMID- 11015929 TI - Sensitivity of nucleus-nucleus cross sections and atomic-electron effects in dissipative heavy-Ion collisions AB - We argue that spontaneous microchannel S-matrix correlations and slow spin decoherence result in a sensitivity of the cross sections for strongly dissipative heavy-ion collisions to an arbitrarily small perturbation. Such a sensitivity implies that atomic electrons should influence energetic ( approximately 100 MeV) heavy-ion reactions. The atomic-electron effects are predicted to be approximately 100% of the magnitude of the non-self-averaged oscillating component of the nucleus-nucleus cross sections. PMID- 11015930 TI - Observation of the continuous stern-gerlach effect on an electron bound in an atomic Ion AB - We report on the first observation of the continuous Stern-Gerlach effect on an electron bound in an atomic ion. The measurement was performed on a single hydrogenlike ion ( 12C5+) in a Penning trap. The measured g factor of the bound electron, g = 2.001 042(2), is in excellent agreement with the theoretical value, confirming the relativistic correction at a level of 0.1%. This proves the possibility of g-factor determinations on atomic ions to high precision by using the continuous Stern-Gerlach effect. The result demonstrates the feasibility of conducting experiments on single heavy highly charged ions to test quantum electrodynamics in the strong electric field of the nucleus. PMID- 11015931 TI - Observation and characterization of the fluorescence decay of the 2s2p(6)np 1P(o) excited states of Ne AB - Fluorescence decay of the 2s2p(6)np (1)P(o) excited states of neon has been observed directly for the first time. This process has also been observed indirectly by threshold photoelectron spectroscopy and in electron/photon coincidences. The measurements have allowed this weak decay channel to be characterized and branching ratios derived. The fluorescence excitation spectrum is well described by a simple Rydberg model that highlights the interplay and competition between the decay channels of autoionization and fluorescence. PMID- 11015932 TI - Photoionization of highly charged ions using an ECR ion source and undulator radiation AB - Photoionization of Xe4+ to Xe7+ ions was studied by combining an electron cyclotron resonance ion source with synchrotron radiation. Multiconfiguration Dirac-Fock calculations were performed to interpret the data. Many autoionization lines were measured and identified, resulting from excitation of a 4d electron into nf and np orbitals followed by Auger decay of the excited states. Continuum photoionization is negligible for the higher members of the isonuclear series. PMID- 11015933 TI - Beyond optical molasses: 3D raman sideband cooling of atomic cesium to high phase space density AB - We demonstrate a simple, general purpose method to cool neutral atoms. A sample containing 3x10(8) cesium atoms prepared in a magneto-optical trap is cooled and simultaneously spin polarized in 10 ms at a density of 1.1x10(11) cm (-3) to a phase space density nlambda(3)(dB) = 1/500, which is almost 3 orders of magnitude higher than attainable in free space with optical molasses. The technique is based on 3D degenerate Raman sideband cooling in optical lattices and remains efficient even at densities where the mean lattice site occupation is close to unity. PMID- 11015934 TI - Recoil-Ion momentum distributions for single and double ionization of helium in strong laser fields AB - We have measured the momentum distributions of singly and doubly charged helium ions created in the focus of 220 fs, 800 nm laser pulses at intensities of (2.9 6.6)x10(14) W/cm(2). All ions are emitted strongly aligned along the direction of polarization of the light. We find the typical momenta of the He2+ ions to be 5 10 times larger than those of the He1+ ions and a two peak structure at the highest intensity. PMID- 11015935 TI - Momentum distributions of ne(n+) ions created by an intense ultrashort laser pulse AB - Vector momentum distributions of Ne(n+) (n = 1,2,3) ions created by 30 fs, approximately 1 PW/cm(2) laser pulses at 795 nm have been measured using recoil ion momentum spectroscopy. Distinct maxima along the light polarization axis are observed at 4.0 and 7.5 a.u. for Ne2+ and Ne3+ production, respectively. Hence, mechanisms based on an instantaneous release of two (or more) electrons can be ruled out as a dominant contribution to nonsequential strong-field multiple ionization. The positions of the maxima are in accord with kinematical constraints set by the classical "rescattering model." PMID- 11015936 TI - New summation rules for coulomb wave functions AB - Sums of products of the Coulomb wave functions over degenerate manifolds have been obtained in a closed form. These sums appear in many atomic and molecular problems. The sums have been obtained making use of the properties of the Coulomb Green's function G(r, r('),E), in the limit E-->E(n), where E(n) is the eigenenergy of the hydrogenlike atomic ion. The closed Hostler-Pratt form of G in the coordinate representation has been used. The sums calculated are a consequence of the n degeneracy of the Coulomb atomic energy levels. This itself, as is well known, follows from the four-dimensional symmetry of the Coulomb problem for the hydrogen atom. PMID- 11015937 TI - Dynamics of solvation-induced structural transitions in mesoscopic binary clusters AB - Ion solvation dynamics following collision of K+ with mesoscopic water-ammonia aggregates is investigated. Two processes determine the postcollision state of the clusters: First, a morphological transformation from a structure characterized by a solid water core and a liquid-phase ammonia sheath into a dumbbell-shaped cluster composed of a liquid water droplet attached to an ammonia subcluster containing the solvated ion. Second, evaporation cools the cluster during and after the morphological change and this evaporative cooling is quantitatively characterized by determining the size-dependent rate constants for these complex clusters. PMID- 11015938 TI - Focusing virtual photons: casimir energies for some pairs of conductors AB - We consider the Casimir energy E of a pair of conductors, E( parallel) for parallel plates, E(|o) for a plate and a sphere, E(oo) for two separated spheres, and E( middle dot in circle) for one sphere inside the other. We also obtain E((o) for an open shell and a sphere, a configuration which might be experimentally preferable. Semiclassically the Casimir energy is given by the optical properties of the system of coaxial mirrors, with focal lengths f(1) and f(2), a distance l<xB--> flow. The neoclassical radial electric field E(r) shows no bifurcation in contrast with earlier orbit loss models, but the shear in E(r) reaches values at which a transition to enhanced confinement has been observed. Also, MHD turbulence analysis shows that a smooth transition can occur through the neoclassical E-->xB--> flow shear suppression. The parameter scaling of threshold temperature for strong turbulence shear suppression agrees with the H-mode threshold scaling in ASDEX Upgrade. PMID- 11015946 TI - Coherent structure phenomena in drift wave-zonal flow turbulence AB - Zonal flows are azimuthally symmetric plasma potential perturbations spontaneously generated from small-scale drift-wave fluctuations via the action of Reynolds stresses. We show that, after initial linear growth, zonal flows can undergo further nonlinear evolution leading to the formation of long-lived coherent structures which consist of self-bound wave packets supporting stationary shear layers. Such coherent zonal flow structures constitute dynamical paradigms for intermittency in drift-wave turbulence that manifests itself by the intermittent distribution of regions with a reduced level of anomalous transport. PMID- 11015947 TI - H-Mode operation in the START spherical tokamak AB - H-mode operation has been achieved in high current (I(p)>200 kA) plasmas in the START spherical tokamak for both neutral-beam-injection-heated and Ohmic discharges. The transition to H mode features the development of well-defined edge pedestals in density and temperature, which signifies the formation of an edge-transport barrier, and associated edge-localized modes. Recent operation at plasma currents exceeding 250 kA shows that these features are accompanied by increases in energy confinement time. This is the first clear demonstration of the H-mode regime in a spherical tokamak. PMID- 11015948 TI - Chain segment order in polymer thin films on a nonadsorbing surface: A NMR study AB - The dynamics of polydimethylsiloxane chains confined between air and a solid, wettable, and nonadsorbing surface is investigated using deuterium NMR. Chain segments in such molten films undergo uniaxial reorientations around the normal to the surfaces. The influence of the film thickness proves this dynamic uniaxiality involves rapid segment diffusion between both surfaces which impose a high degree of planar orientation. This segmental ordering is observed over a thickness range which depends on the degree of chain entanglements. PMID- 11015949 TI - Stability of Si-interstitial defects: from point to extended defects AB - Trends in the growth of extended interstitial defects are extracted from extensive tight-binding and ab inito local density approximation simulations. With an increasing number of interstitials, the stable defect shape evolves from compact to chainlike to rodlike. The rodlike 311 defect, formed from (011) interstitial chains, is stabilized as it grows, elongating in the chain direction. Accurate parametrization of the defect-formation energy on the number of interstitials and interstitial chains, together with the anisotropy of the interstitial capture radius, enables macroscopic defect-growth simulations. PMID- 11015950 TI - Effect of defects on the elastic properties of Wustite AB - Wustite (FexO) has been studied under static high pressure in the composition range x = 0.95-0.99. Fe0.99O is substantially less compressible than FexO with x<0.98. Such observations provide the only direct evidence of a significant effect of defect concentration on the elastic properties and support the results of crystal-field modeling for ideal stoichiometric wustite. The bulk moduli of FexO show a discontinuous variation at compositions that are within the predicted range for the defect clusters to form, a behavior that may be caused by an order disorder transition of the defect structure. PMID- 11015951 TI - Stokes formula and density perturbances for driven tracer diffusion in an adsorbed monolayer AB - We study the intrinsic friction of monolayers adsorbed on solid surfaces from a gas phase or vapor. Within the framework of the Langmuir model of delocalized adsorption, we calculate the resistance offered by the mobile adsorbate's particles to some impure tracer molecule, whose diffusive random motion is biased by a constant external force. We find that for sufficiently small driving forces the force exerted on the tracer shows viscouslike behavior. We derive then the analog of the Stokes formula for two-dimensional adsorbates, calculate the corresponding friction coefficient, and determine the stationary particle distribution in the monolayer as seen from the driven impurity. PMID- 11015952 TI - Strongly temperature dependent sliding friction for a superconducting interface AB - A sudden drop in mechanical friction between an adsorbed nitrogen monolayer and a lead substrate occurs when the lead passes through the superconducting transition temperature. We attribute this effect to a sudden drop at the superconducting transition temperature of the substrate Ohmic heating. The Ohmic heating is due to the electronic screening current that results from the sliding adsorbed film. PMID- 11015953 TI - Direct measurement of interfacial curvature distributions in a bicontinuous block copolymer morphology. AB - Self-consistent field theory predicts that the complex phase behavior of block copolymers does not originate solely from the interface seeking constant mean curvature as once thought, but instead reflects competing minimization of interfacial tension and packing frustration. To test this prediction, we directly measure interfacial curvature distributions from a 3D image reconstruction of the bicontinuous gyroid morphology. Results obtained here reveal that the gyroid interface is not constant mean curvature and confirm the importance of packing frustration in the stabilization of such complex nanostructures. PMID- 11015954 TI - Spectral weight of the hubbard model through cluster perturbation theory AB - We calculate the spectral weight of the one- and two-dimensional Hubbard models by performing exact diagonalizations of finite clusters and treating intercluster hopping with perturbation theory. Even with relatively modest clusters (e.g., 12 sites), the spectra thus obtained give an accurate description of the exact results. Spin-charge separation (i.e., an extended spectral weight bounded by singularities dispersing with wave vector) is clearly recognized in the one dimensional Hubbard model, and so is extended spectral weight in the two dimensional Hubbard model. PMID- 11015955 TI - Enhanced susceptibility in LNiO3 perovskites ( L = La,Pr,Nd,Nd0.5Sm0. 5) AB - The temperature dependence of the resistivity rho(T) and of the dc magnetic susceptibility chi(T) were measured on high-quality LNiO3 (L = La,Pr,Nd,Nd0.5Sm0.5) samples synthesized under high oxygen pressure. Subtraction of the rare-earth contribution to chi(T) allows the presentation of the evolution of the susceptibility of the NiO3 array from Pauli to Curie-Weiss paramagnetism with decreasing bandwidth. A metal-insulator transition occurring at a temperature T(t) = T(N) is first order for L = Pr and Nd; it becomes second order and produces no anomaly in chi(-1)(T) at a T(t)>T(N) for L = Nd0.5Sm0.5. In the antiferromagnetic domain T or = 11.0 ng/mL in 36 patients (53.73%). CEA was helpful in the diagnosis in 18 cases, increasing to 57 (85.07%) the number of positive diagnoses. Eight samples of nonepithelial tumors had low levels of CEA. In 55 patients with nonneoplasic ascites the cytopathologic examination was negative, but the CEA assay was > 11.0 ng/mL in 3 patients. CONCLUSION: The cytopathologic examination should be performed in all cases, and the CEA assay should be done in suspected cases of epithelial neoplasia in which the cytologic examination was negative, there was uncertainty about the histologic type of neoplasia, or a diagnosis of nonepithelial neoplasia was made. When ascitic leukocytosis or hepatic failure is present, one should be cautious in interpreting the CEA assay because false positivity can occur. PMID- 11015981 TI - Phyllodes tumor of the breast. A cytohistologic study of 80 cases. AB - OBJECTIVE: To study the cytologic features of phyllodes tumor (PT) of the breast and determine the accuracy of their subclassification in fine needle aspirates. STUDY DESIGN: Eighty cases of histologically diagnosed PT between 1982 and 1997 with a previous fine needle aspiration (FNA) were evaluated. The FNA smears of each case were reviewed without knowledge of the initial cytologic diagnosis and subclassified into benign, borderline or malignant PT. RESULTS: Benign PTs were characterized by a dimorphic mixture of stromal and epithelial cells. The stromal fragments showed mild to moderate cellularity with absent to minimal pleomorphism and no mitosis. There were occasional, if any, single stromal cells. Borderline PTs had stromal fragments with moderately cellular stroma exhibiting moderate pleomorphism. Two additional features were the presence of single stromal cells and an occasional mitosis in the stromal fragments/single cells. Aspirates from malignant PT were very cellular, with a high stromal/epithelial ratio and marked stromal cellularity. The stromal cells were highly pleomorphic, with frequent mitosis and atypical single stromal cells in the background. Fifty-seven of the 80 histologically documented cases (71.3%) were diagnosed as PT on FNA (40 benign, 10 borderline and 7 malignant). In 81% (46 of 57 PTs), good cytohistologic correlation (32 benign, 8 borderline and 6 malignant) was observed. In another eight cases, one grade differentiation between cytologic and histologic grade was observed. Six of the nine malignant PTs on histology were correctly subclassified on cytology. There were one false positive and two false negative cases. CONCLUSION: Cytologic diagnosis and grading of PT on FNA is possible. Special care should be undertaken in interpreting phyllodes fragments, cellularity of stroma, pleomorphism and mitosis. Single stromal cells are also important morphologic criteria for subclassification. Multiple-site aspiration is advisable to avoid diagnostic errors. PMID- 11015980 TI - Peritoneal washings in endometrial carcinoma. A study of 298 patients with histopathologic correlation. AB - OBJECTIVE: To correlate findings of peritoneal washings in patients with endometrial carcinoma with histologic parameters. STUDY DESIGN: Between 1995 and 1998, 298 women with endometrial carcinoma were treated by hysterectomy with intraoperative peritoneal washings (PW) at Memorial Sloan-Kettering Cancer Center. All cytology and pathology slides were available for review. Pathologic parameters of hysterectomy specimens were evaluated and correlated with the findings of PW. RESULTS: Thirty-two patients (10.7%) had abnormal PW. Two hundred sixty-two had endometrioid adenocarcinoma; 26 of them had abnormal PW (10.0%). Thirty-six patients had other histologic subtypes (papillary serous carcinoma, clear cell carcinoma and adenosquamous carcinoma), and six of them had abnormal PW (16.7%). The incidence of abnormal PW in the two groups was not significantly different (P = .78). Among 26 patients with endometrioid adenocarcinoma and abnormal PW, there were 17 cases (9.9%) of International Federation of Gynecology and Obstetrics (FIGO) grade 1, 7 (12.7%) of grade 2 and 2 (5.7%) of grade 3 (P = .56). Ten cases (14.9%) had no myometrial invasion, 10 (7.0%) had myometrial invasion of < or = 50% of myometrial thickness, and 6 (11.5%) had invasion of > 50% of myometrial thickness (P = .18). Vascular invasion was present in 8 cases (14.8%) and absent from 17 (8.2%) (P = .14). Eighteen patients (7.6%) had stage I/II disease, and eight patients (30.8%) had stage III/IV disease (P = .001). Among 298 patients, cervicovaginal smears performed before surgery were available for review in 76. Five of the 7 patients (71.4%) with abnormal PW and 37 of the 69 patients (53.6%) with normal PW had abnormal Pap smears (P = .45). CONCLUSION: Abnormal PW did not correlate with histologic subtypes, FIGO grade, depth of myometrial invasion, vascular invasion or abnormal Pap smears. A significantly higher incidence of abnormal PW was associated with stage III/IV disease. PMID- 11015982 TI - Esophageal exfoliative cytology samplers. A comparison of three types. AB - OBJECTIVE: To compare three types of esophageal exfoliative cytology samplers in terms of patient acceptability, ease of use, final destination of the sampler tip in the gastrointestinal tract and cellular yield. STUDY DESIGN: A controlled, single-blind, cross-over study was undertaken to compare the balloon, sponge and sponge-mesh samplers in healthy volunteers. After completing the three procedures in random order, participants were asked about their preferred method. Ease of use was defined as the technician's ability to perform the intubation successfully. Final destination of the samplers was assessed fluoroscopically. Cytopathologists determined the cellular yield of each sampler using the Bethesda System. RESULTS: Sixty-two volunteers participated. The two encapsulated samplers were significantly preferred over the balloon (P < .0001). There was no significant difference in ease of use, final destination or cellular yield of the three techniques. All three samplers were successfully intubated on the first attempt and retrieved adequate numbers of squamous and glandular cells in > 78% of cases. CONCLUSION: All three samplers obtained satisfactory yields of squamous and glandular cells, but the encapsulated samplers were more patient acceptable. The sponge-mesh sampler may be the least complicated sampler for field screening use. Larger-scale studies will be required to test the accuracy of these three samplers for detecting esophageal dysplasia and carcinoma. PMID- 11015983 TI - Langerhans cell histiocytosis of the thyroid gland. A case report. AB - BACKGROUND: The variant of Langerhans histiocytosis commonly encountered in adults is the benign, localized form (eosinophilic granuloma). The more aggressive or diffuse type (Letterer-Siwe disease) is rare in adults. CASE: A 28 year-old woman presented with enlargement of the thyroid gland three years after she had been diagnosed with and placed on treatment for diabetes insipidus. Thyroidectomy was performed following an initial fine needle aspiration cytology report of either papillary thyroid carcinoma or Langerhans cell histiocytosis. The latter diagnosis was confirmed on histopathology and immunohistochemical staining for S-100 protein. Intracellular Birbeck granules were also demonstrated by electron microscopy. The disease progressed over a 10-week period to involve the kidneys, resulting in renal dysfunction. CONCLUSION: Synchronous or metachronous involvement of the hypothalamus and thyroid gland by Langerhans histiocytosis could not be excluded in the present case, with subsequent progression to involve other organs. This was an unusual presentation of the disease in an adult. PMID- 11015984 TI - Fine needle aspiration cytology of follicular dendritic cell sarcoma. Report of a case with cytologic detection in an extranodal site. AB - BACKGROUND: Follicular dendritic cell sarcoma (FDCS) is a rare malignancy of accessory immune cells that can present in both nodal and extranodal sites. Previous cytologic case reports of FDCS have focused on fine needle aspiration (FNA) findings in nodal sites with low grade morphology and indolent clinical courses. CASE: A 33-year-old female presented with a three-month history of abdominal distention, early satiety and nausea. Initial imaging studies showed a large abdominal mass, with subsequent studies showing lung, liver and lymph node metastases. Examination of primary and metastatic tumors by a combination of conventional histology, immunohistochemistry and FNA demonstrated an extranodal intraabdominal follicular dendritic cell sarcoma. CONCLUSION: FDCS demonstrates a characteristic cytologic picture on FNA, with important cytologic features, including both syncytial and discohesive large epithelioid to spindled malignant cells with intranuclear inclusions, nuclear grooves and a prominent, mature, lymphocytic inflammatory component. No evidence of morphologic tumor progression was noted in comparison of primary and metastatic tumors. To aid in the cytologic distinction of FDCS from other similar-appearing neoplasms, we recommend acquisition of material for immunohistochemical studies, recognition of diverse clinical presentations (including extranodal and aggressive) and acknowledgment of the range of tumor morphologic grades. PMID- 11015985 TI - Cryptococcal osteomyelitis. Report of a case with aspiration biopsy of a humeral lesion with radiologic features of malignancy. AB - BACKGROUND: Osteomyelitis due to Cryptococcus neoformans typically exhibits lytic lesions on radiographs. Extensive periosteal reaction is an uncommon feature. CASE: A 68-year-old man presented with pain and swelling in the left elbow. Radiologic studies exhibited a lytic humeral lesion with extensive periosteal reaction, interpreted as a malignant neoplasm. Fine needle aspiration biopsy (FNA) revealed abundant cryptococcal organisms. CONCLUSION: Cryptococcus is an uncommon cause of lytic osseous lesions that may mimic malignant neoplasms. Extensive periosteal reaction may support a radiologic diagnosis of primary osseous malignancy in rare cases. FNA with examination of Diff-Quik-stained slides may be employed for distinguishing cryptococcal osteomyelitis from malignant tumors and for prompt identification of the organisms. PMID- 11015986 TI - Pediatric acute leukemia presenting as bilateral renal enlargement. Report of a case with fine needle aspiration cytologic features suggestive of megakaryocytic differentiation. AB - BACKGROUND: Fine needle aspiration (FNA) cytologic diagnosis of acute myeloid leukemia involving the kidney has rarely been reported, but acute leukemia with cytologic features suggestive of megakaryocytic differentiation has not been described before. CASE: An 8-month-old male presented with an abdominal swelling, bilateral cervical and inguinal lymphadenopathy and enlarged left epididymis. Ultrasonography of the abdomen revealed a space-occupying lesion in the liver and bilateral enlargement of the kidneys. FNA smears from the right kidney and right submandibular lymph node showed numerous blast cells. Since rare blast cells were positive for myeloperoxidase, a cytodiagnosis of involvement by acute myeloid leukemia (AML) was made. However, following the hematologic diagnosis of acute megakaryoblastic leukemia (M7) from peripheral blood and bone marrow smear examination, FNA smears were reviewed. There were cytoplasmic blebs or protrusions in the blast cells and cytologic features suggestive of their differentiation toward micro-megakaryocytes and megakaryocytes. There was also evidence of shedding of platelets, including numerous giant platelets. The reviewed FNA cytodiagnosis was suggestive of AML (M7). CONCLUSION: Extramedullary involvement by acute megakaryoblastic leukemia (M7) can be suspected based on cytomorphologic features in FNA smears. PMID- 11015987 TI - Signet ring cell lobular carcinoma of the breast presenting in a cervicovaginal smear. A case report. AB - BACKGROUND: The presence of extragenital malignant cells in cervicovaginal smears is a rare and usually late event in a patient with a long history of cancer. This, to the best of our knowledge, is the first case of breast cancer initially diagnosed on a Pap smear. CASE: A 50-year-old woman presented with abdominal distension and weight gain. A Pap smear showed numerous signet ring cells and was diagnosed as signet ring cell adenocarcinoma, most likely metastatic from the breast or stomach. Subsequent evaluation revealed bilateral adnexal masses and inguinal lymphadenopathy, leading to hysterectomy and bilateral salpingo oophorectomy. The ovaries, corpus and cervix were involved by signet ring cell carcinoma. The metastatic tumor proved to be positive for CK7, CEA, estrogen receptor and progesterone receptors and negative for CK20. Despite the absence of a discrete palpable breast mass, a mammogram was recommended based on these results, and an ensuing breast biopsy showed the presence of an infiltrating lobular carcinoma. CONCLUSION: Lobular carcinoma of the breast may present in a cervicovaginal smear. Correct interpretation of signet ring cells as metastatic, most likely from the breast or stomach, is helpful in guiding management. PMID- 11015988 TI - Fine needle aspiration cytology of adenocarcinoma of the rectovaginal septum. A case report. AB - BACKGROUND: Adenocarcinoma arising in the rectovaginal septum is exceedingly rare and is difficult to diagnose by pathologic examination prior to surgery because of the anatomic position of the tumor. CASE: A 42-year-old woman presumed to have adenocarcinoma of the rectovaginal septum underwent fine needle aspiration for diagnosis. Although a previously performed biopsy from the posterior vaginal fornix was unsuccessful, fine needle aspiration cytology via the posterior vaginal wall detected adenocarcinoma cells. The cell clusters were composed of cells with enlarged and hyperchromatic nuclei. The nuclei themselves demonstrated round and/or irregular morphologic patterns, with high nuclear/cytoplasmic ratios, and often contained an enlarged, round nucleolus and sometimes multiple ones in a single nucleus. Aniso-nucleosis was severe, and the chromatin patterns ranged from coarse to finely granular. The cytoplasm was narrow and lightly stained. Following fine needle aspiration, the patient underwent posterior exenteration on the basis of the cytologic diagnosis. CONCLUSION: Fine needle aspiration cytology was useful in establishing the preoperative diagnosis of adenocarcinoma of the rectovaginal septum, and curative exenterative surgery could be then performed. To our knowledge, this is the first report of fine needle aspiration cytology of adenocarcinoma at this location. PMID- 11015989 TI - Cytologic diagnosis of leishmaniasis in HIV infection. A report of eight cases. AB - BACKGROUND: Leishmania organisms are among the intracellular microorganisms with a tendency to develop in patients with the acquired immunodeficiency syndrome (AIDS). With increasing travel to endemic areas by patients with human immunodeficiency virus (HIV) infection, it is becoming a more-frequent diagnosis in nonendemic areas. CASES: Ten cytologic specimens from eight patients with leishmaniasis and AIDS were reviewed. Eight samples were obtained from lymph nodes through fine needle aspiration (FNA). Another sample was obtained after scraping a tongue ulcer. The last one was an ascitic fluid specimen. Smears showed numerous parasitized histiocytes with abundant intracellular Leishmania organisms (amastigotes). Extracellular microorganisms were also abundant. Diff Quik-stained smears allowed the clear recognition of the characteristic morphologic appearance with a deep-staining area (nuclei) and paranuclear zone (kinetoplast). Intracellular organisms were round, while single, extracellular forms were a more elongated. CONCLUSION: The polymorphous clinical manifestations usually seen in patients suffering from leishmaniasis and AIDS constitute a diagnostic challenge that can be facilitated by cytopathologic examination. Cytology permits easy and rapid identification of Leishmania amastigotes, allowing a specific diagnosis and treatment. PMID- 11015990 TI - [Specific immunotherapy with cancer vaccines]. AB - With the recent progress in molecular biology and gene technology, many new cancer-specific antigens have been identified. Many studies have demonstrated the role of HLA class I-restricted cytotoxic T lymphocytes (CTLs) in cancer specific immunotherapies. We have also established HLA-A24- and A26-restricted and cancer specific CTLs from a patient with squamous cell carcinoma of the esophagus. Using CTLs, we identified a new gene SART-1 by cDNA-expression cloning and some SART-1 derived cancer rejection peptides were also identified. Further more, using the same approach, we identified a cyclophilin B gene that encodes antigenic epitopes recognized by HLA-A24-restricted and tumor-specific CTLs. Now we are performing phase I trials using these peptide vaccines and have found an increase in CTL precursor frequency in some cases in an in vitro study. However, other recent studies have reported that many tumors escape from CTL recognition by downregulation of HLA class I expression. Moreover, most cancer cells produce a suppressor agents against the immune system. Therefore, we must resolve these major problems to produce successful cancer vaccine therapy soon. PMID- 11015991 TI - [Up to date of multidisciplinary treatments centering around hepatectomy for advanced liver cancer in stage IV-A]. AB - Patients with advanced Stage IV-A primary liver cancer, hepatocellular carcinoma (HCC) can be divided into subgroups: those with involvement of a major branch of the portal (Vp3) or hepatic (Vv2, Vv3) veins and those having multiple tumors in both lobes without Vp3 or Vv2, Vv3. The prognosis of Stage IV-A patients with Vv2 or Vv3 may be improved by extended hepatectomy with resection and reconstruction of hepatic veins or IVC. In those with Vp3, multidisciplinary treatments consisting of extended hepatectomy and adjuvant chemotherapy, i.e. intra-arterial injection or TACE, are thought to be feasible at the present, but the outcomes are still poor. On the other hand, there are some Stage IV-A patients with multi centrical tumors who have multiple tumors in both lobes without major vascular invasion, and their prognoses are improved by partial resection of each tumor. However, when there are multiple tumors caused by intrahepatic metastases, multidisciplinary treatments consisting of reduction surgery, microwave ablation, ethanol injection, and intra-arterial chemotherapy might be useful at present. PMID- 11015992 TI - [Percutaneous tumor ablation therapy for the advanced stage of HCC]. AB - We have performed percutaneous tumor ablation (PTA) including percutaneous ethanol injection therapy (PEIT) for 90% of the patients with hepatocellular carcinoma. Until December 1998, the 793 patients received PTA, 5 years survival rate reached 39.8%. Excluding the patients with Child C whose hepatic function were extremely low, 5 years survival rate reached to the level of 41.2%. Since 5 years survival rate in stage IV-A reached 24.4%, the patients of stage IV-A may be considered to have an indication for PTA. We have confirmed the effectiveness of the local treatment including radiotherapy for advanced hepatocellular carcinoma with portal vein invasion. We are attempting to perform PTA for the extra-hepatic lesions that had no indication of other treatment. However the indication of PTA is limited by the presence of diffuse nodules, exacerbation of the hepatic function, or tumor invasion to portal vein, bile duct, inferior vena cava. PMID- 11015993 TI - [Indication and limitation of microwave coagulation therapy for stage IV-A hepatocellular carcinoma]. AB - Patients with stage IV-A hepatocellular carcinoma have been considered to have dismal prognosis. However, among them there are many of patients with multiple carcinoma of multicentric origin, whose prognosis is not necessarily poor. For treatment of multiple carcinoma, preoperative evaluation for the malignant aggressiveness of each nodule, and for liver function impairment is essential. Although hepatic resection has a high curability, resection sometimes impair long term survival because of postoperative deterioration of liver function. In this point, heat ablation therapy such as microwave coagulation therapy is favorable treatment, and for some nodules at early stage heat ablation therapy would have a sufficient curability to achieve long term survival. Authors decide on the selection of treatment (resection or heat ablation therapy) according to preoperative evaluation for the malignant aggressiveness by diagnosis of tumor gross type, and for the degree of liver impairment by serum hyaruronic acid level. PMID- 11015994 TI - [Transcatheter arterial embolization for advanced hepatocellular carcinoma- indications and limitations]. AB - Many patients with advanced hepatocellular carcinoma (HCC) in stage IV have no surgical indications. Transcatheter methods such as transcatheter arterial embolization (TAE) and hepatic arterial infusion chemotherapy play a main role of the treatment for advanced HCC. Conventional TAE (from proper hepatic artery) is performed for patients without liver dysfunction. Patients with severe liver dysfunction could not in the past be treated with TAE, but lately it has become possible to treat them with the method of segmental TAE or subsegmental TAE due to the development of a microcatheter and advances in equipment. Although technical progress is remarkable, there are no fixed guidelines for advanced HCC. Suitable methods for individuals need to be discussed. PMID- 11015995 TI - [Hepatic arterial infusion chemotherapy using totally implantable port system]. AB - Because of low grade of response rate and high grade of toxicity, arterial infusion chemotherapy (AIC) is not the first choice therapy for hepatocellular carcinoma (HCC). Only far advanced HCC patients, who have no indication of surgical, transcatheter arterial embolization nor ablation therapy, are able to accept AIC. The AIC using totally implantable port system has an advantage for patient's quality of life in the long term. We recommend new intensive EEP regimen which has the response rate of 43% although its severe toxicity. PMID- 11015996 TI - [Long-term outcome of preoperative chemotherapy with 5'-deoxy-5-fluorouridine (5' DFUR) for gastric cancer]. AB - We performed a multicenter clinical trial of preoperative chemotherapy for gastric cancer. Patients aged 75 years or less with advanced gastric cancer were enrolled and randomized into the following groups: Group I, which received oral 5'-DFUR (610 mg/m2/day x 10 days or over) preoperatively, and Group II, which received no treatment preoperatively. Patients in both groups also received intravenous MMC 1 and 2 days after surgery and were orally administered 5'-DFUR for two years postoperatively. There were 171 patients (Group I: 91, Group II: 80) enrolled and analyzed, and the 5-year survival rate was 63.4% in Group I and 64.9% in Group II (p = 0.698). Among patients classified as having curability B, the 5-year survival rate of each group was 51.8% and 36.8%, respectively (p = 0.426). However, the 5-year survival rate of patients showing good compliance with oral 5'-DFUR therapy was significantly higher than that of patients with poor compliance (53.3% vs 22.0%, p = 0.010). The pyrimidine nucleoside phosphorylase (PyNPase) activity in gastric carcinoma tissue from Group I was lower than that from Group II, and Group II patients tended to die of hematogeneous metastases. In conclusion, although this clinical trial failed to demonstrate a significant benefit of preoperative chemotherapy because of the low dose of 5'-DFUR, we believe that good compliance with oral anticancer treatment contributes to long-term survival and that 5'-DFUR reduces hematogeneous metastasis of gastric carcinoma. PMID- 11015997 TI - [5-FU concentration in the serum and the tumor tissue after administration of UFT 200 mg/day to patients over 80 years of age with oral cancer]. AB - UFT was administered orally at a dosage of 200 mg/day, 2 times a day, to patients over 80 years of age with oral cancer. The concentration of 5-FU in the serum and tumor tissue, as well as the side effects, were investigated. The results were as follows: 1. The concentration of 5-FU in the serum peaked (0.017 to 0.066 microgram/ml) 1 or 2 hours after UFT administration. The concentration 8 hours after administration was relatively high (0.016 to 0.041 microgram/ml). 2. The 5 FU concentrations in the tumor tissues in 3 out of 5 cases were greater than 0.05 microgram/g, which is considered to be the effective level. The concentration tended to be higher with increased duration of administration. 3. A minor side effect, bone marrow dysfunction, was observed. No effect on the function of the liver or digestive system was observed. PMID- 11015998 TI - [Significance of interferon alpha therapy for advanced renal cell carcinoma. Fukushima Renal Cancer Study Group]. AB - We ranked prognostic factors to retrospectively evaluate the clinical significance of interferon alpha (IFN-alpha) therapy in patients with Robson stage IVB renal cell carcinoma. A total of 44 Robson stage IVB renal cancer patients were divided into 2 groups, one with more than 6 months administration of IFN-alpha (3-7 times a week: group A) and another without any IFN-alpha administration. The distribution of these 2 groups was not randomized. In addition to IFN-alpha therapy, survival was analyzed with respect to performance status (PS), mass reductive nephrectomy, concomitant use of other cytotoxic therapies, the number of metastatic organs, growth type, site of metastasis and the period of diagnosis, using a multivariate method with Cox proportional hazards regression. The multivariate analysis showed administration of IFN-alpha to be the most significant factor influencing a good prognosis. Improved survival was also significantly correlated with slow growing type and good PS. Among group A, a significant favorable prognosis was obtained in patients with the responses of no change (NC), partial response (PR) and complete remission (CR) 6 months after initiating administration of IFN-alpha, as well as with good PS and a slow growing type carcinoma. We conclude that IFN-alpha therapy might improve the prognosis of patients with Robson stage IVB renal cell carcinoma, especially, in cases when a greater than NC response is obtained after 6 months administration of IFN-alpha. PMID- 11015999 TI - [Long-term follow up of chronic myelogenous leukemia patients treated with natural interferon alpha--multi-institutional cooperative study]. AB - The effects on survival in patients with chronic myelogenous leukemia (CML) treated with natural interferon alpha (Sumiferon) were analyzed. The subjects were 131 patients with CML who underwent treatment with Sumiferon between August 1991 and December 1992. Sumiferon was administered more than 8 weeks, and patients were followed for 5 years. In the end, 101 patients were analyzed after 30 patients were dropped from the study because they had received bone marrow transplantation. Survivals from the start of Sumiferon administration were 63.4% at 3 years, 52.5% at 4 years, and 42.6% at 5 years, respectively. Survivals were compared between the 74 patients who received Sumiferon administration with 1 year of diagnosis, and the 27 patients who received Sumiferon administration after more than 1 year from diagnosis. The 50% survivals were 2,089 days and 868 days, respectively (p = 0.0011). It was concluded that early administration of interferon alpha results in a prolonged survival of CML patients. PMID- 11016000 TI - [High-dose chemotherapy with autologous peripheral blood stem cell transplantation for treatment of non-Hodgkin's lymphoma]. AB - Findings for 41 patients with non-Hodgkin's lymphoma (NHL) treated with high-dose chemotherapy (HDC) and/or autologous peripheral blood stem cell transplantation (PBSCT) are reported. Two of the 41 patients were treated with HDC alone without PBSCT. At transplant, 20 patients were in complete remission, while 19 had resistant NHL and had failed to achieve a complete remission (CR) after several courses of conventional chemotherapy. The conditioning regimens used were mainly ACE (cytarabine, cyclophosphamide, etoposide) and MEAC (MCNU, etoposide, cytarabine, cyclophosphamide). The treatment-related mortality rate was 4.9%. Two patients treated with MEAC died from intractable congestive heart failure. Nine of the 19 patients with resistant NHL achieved CR, and at a median follow-up of 26 months (range, 3 to 93 months) the estimated two-year disease-free survival rate for these patients was 44.4%. Four patients in CR at present were in partial remission before HDC and PBSCT. Fifteen of the 20 patients in CR before HDC were transplanted in first CR and 5 in 2nd CR. At a median follow-up of 49 months (range, 3 to 96 months), the estimated 3-year DFS for the group of all patients was 73.7%. Five relapses occurred between 5 and 35 months post-transplantation. In conclusion, HDC and PBSCT as induction therapy was only effective for patients with resistant NHL who responded to conventional chemotherapy, and may improve the survival of patients in CR as consolidation therapy. PMID- 11016001 TI - [A randomized crossover comparison of azasetron alone and azasetron plus dexamethasone for the prevention of nausea and vomiting by chemotherapy including cisplatin]. AB - A randomized crossover study between azasetron alone and azasetron combined with dexamethasone was performed to investigate the prevention of nausea and vomiting due to chemotherapy including cisplatin in patients with advanced head and neck carcinoma. The results indicated that the combination therapy with dexamethasone was more effective than azasetron alone in preventing nausea on days 1 through 5 after the administration of cisplatin and was significantly superior to azasetron alone on days 2 and 3. In addition, complete response (no vomiting) and the antiemetic and antinausea efficacy of combination therapy with dexamethasone on day 2 was significantly superior compared to azasetron alone. No adverse effect from either antiemetic therapy was observed in this study. PMID- 11016002 TI - [A case of non-small-cell lung cancer successfully treated using combination chemotherapy with CDDP and vinorelbine]. AB - A 67-year-old woman presented to our hospital with a chief complaint of bloody sputum. A plain chest X-ray a CT scan revealed a tumor shadow 3 cm in size in the middle lobe of the right lung, multiple nodular shadows in the bilateral lung fields and enlarged hilar and mediastinal lymph nodes. A tumor biopsy done under bronchoscopy revealed poorly differentiated adenocarcinoma of the lungs (cT2N3M1). She was given two courses of combination therapy consisting of cisplatin (80 mg/m2) and vinorelbine (20 mg/m2). The primary tumor in the middle lobe of the right lung and the lung metastases were markedly reduced in size, and a complete response was obtained. The only adverse events were grade 4 neutropenia and grade 2 nausea and vomiting. PMID- 11016003 TI - [A case of bronchial stump recurrence of lung cancer successfully treated with chemoradiotherapy using cisplatin plus vinorelbine]. AB - A 61-year-old man, who had undergone right upper lobectomy for lung cancer about nineteen months earlier, was examined at our hospital for hemoptysis. A bronchial stump recurrence was observed by chest CT and bronchoscopy. The patient underwent chemoradiotherapy using cisplatin (CDDP) and vinorelbine (VNB). Irradiation was started (total 60 Gy/30 fr/6 weeks) about four weeks before the onset of chemotherapy. Chemotherapy consisted of 2 cycles of CDDP 80 mg/m2 on day 1 and VNB 15 mg/m2 on day 1, 8. The dose of VNB was increased to 25 mg/m2 during cycle 2 because cycle 2 was without radiotherapy. After the chemoradiotherapy, the recurrent tumor disappeared on a CT scan and bronchoscopy. Chemoradiotherapy using CDDP plus VNB regimen is considered effective, without serious side effects for advanced non-small-cell lung cancer. PMID- 11016004 TI - [A case of postoperative lung and pleural recurrence after 20 years in a breast cancer patient with marked response to orally administered UFT and cyclophosphamide]. AB - A 62-year-old woman who underwent standard radical mastectomy for left breast cancer developed pleural and pulmonary metastases 16 years later. She complained of slight dyspnea, and computed tomography of the chest revealed a tumor with marked pleural effusion. Her serum level of carcinoembryonic antigen was also increased (49 ng/ml). Systemic chemotherapy with cyclophosphamide, adriamycin, and 5-fluorouracil (CAF) was performed, but she did not show a marked response to CAF therapy. Therefore, she was orally administered UFT and CPA for 5 consecutive days followed by 2 drug-free days. Pleural disseminated foci and a lung tumor were reduced in size on chest CT and the CEA level was decreased within the normal limit. Recently, it was reported that UFT demonstrated significant antiangiogenic activity. This effect may play an important role in the efficacy seen in this patient. PMID- 11016005 TI - [A case of effective chemotherapy using CAF followed by docetaxel for advanced breast cancer]. AB - A huge mass measuring 13 x 12 cm and wide cutaneous edema were detected in the right breast of a 51-year-old woman. Under a diagnosis of locally advanced breast cancer (T4bN2M1, stage IV) with liver metastases, we attempted sequential neoadjuvant chemotherapy. After three courses of CAF therapy (cyclophosphamide, doxorubicin (DXR), 5-FU), the primary tumor was decreased by 56% and the liver metastases had disappeared. A minor pathologic response was observed. Subsequently, three courses of docetaxel (TXT) administration were carried out. The primary tumor was then decreased by 75% and the axillary metastases had disappeared. Histopathological examination showed gross viable tumor cells in the residual tumor and positive axillary lymph nodes. The only toxic effect was nausea (grade 1) and no major adverse effects were observed. Neoadjuvant chemotherapy with sequential DXR followed by TXT is a useful treatment for locally advanced breast cancer. PMID- 11016006 TI - [Weekly chemotherapy with alternating low-dose CPT-11 and low-dose FP against a far advanced pancreatic cancer--a case report]. AB - We attempted a new regimen of weekly chemotherapy with alternating low-dose CPT 11 and low-dose FP against a case of far advanced pancreatic cancer with invasion to the super mesentric artery, celiac artery, and left renal vein. CPT-11 was administered at 25 mg/m2/day x 3/week, and CDDP and 5-FU were administered at 7 mg and 350 mg/m2/day x 3/week by intravenous infusion. These regimens were alternated every week. The serum CA19-9 level decreased gradually with a half time of approximately 2 months (from 972 to 126 U/ml). The tumor was reduced more than 50% 4 months after chemotherapy, and continuing decreasing for more than 6 months. There were no adverse effects except mild leukopenia (less than Grade 1). These results suggest that the combination of low-dose chemotherapy with fewer side effects may be effective not only for tumor shrinkage, but also for a prolonged time to progression. PMID- 11016007 TI - [A case of advanced rectal cancer with unresectable liver metastasis for which chemotherapy was markedly effective]. AB - We report a case of advanced rectal cancer accompanied by unresectable liver metastasis in which remission has been achieved for 7 years. The patient was a 64 year-old woman, and her chief complaints were a feeling of abdominal distention and melena. After low anterior resection (D3), oral administration of 600 mg/day of UFT-E and chemotherapy against the liver metastasis by hepatic arterial injection via a reservoir were started. Since the arterial injection via a reservoir became impossible due to infection, her clinical course was observed solely with oral administration of UFT-E. As a result, complete remission was achieved 7 months postoperatively, and the patient is still alive in good health without recurrences at present (7 years postoperatively). The reason why a favorable outcome has been achieved is probably that, although the initial operation resulted is non-curative resection, measures sufficient to enable the patient to lead an ordinary social life were taken at the beginning, and efforts to achieve a long-term survival were made while respecting the quality of life of the patient so that the patient would be able to coexist with the cancer. PMID- 11016008 TI - [A case report of unresectable colon cancer that disappeared following pharmacokinetic modulation chemotherapy (PMC)]. AB - We encountered a case of unresectable colon cancer that disappeared following pharmacokinetic modulating chemotherapy (PMC). A 41-year-old male underwent ileotransverse anastomosis following a diagnosis of ileus due to unresectable colon cancer with peritoneal dissemination and multiple liver metastases. He was treated weekly with an intravenous infusion of 5-FU, 1,000 mg/body/24 hours and an oral administration of UFT 400 mg/day (PMC). After 6 months of weekly treatment of PMC, colon tumor disappeared and metastatic liver tumors decreased in size. PMC can be administered to outpatients without serious side effects; thus, this therapy improves the prognosis and the patient's quality of life. PMID- 11016009 TI - [A case of colonic cancer with multiple liver metastases effectively treated by intra-arterial chemotherapy]. AB - A 36-year-old female underwent sigmoidectomy and insertion of an intra-hepatic arterial catheter for advanced sigmoid cancer with multiple liver metastases. After the operation, intra-hepatic arterial infusion of the chemotherapeutic agents that showed sensitivity in the histoculture drug response assay (HDRA) to the liver metastasis was done for one year. The metastatic liver lesions other than those in the lateral segment which were fed by an accessory artery decreased remarkably. Therefore, we performed lateral segmentectomy. The patient has been doing well without recurrence for 18 months after the first operation. Intra hepatic arterial infusion chemotherapy for liver metastasis from colorectal cancer can produce an excellent result with the use of sensitive chemotherapeutic agents. PMID- 11016010 TI - [Accelerated titration design]. AB - To reduce the number of patients treated at low and biologically inactive doses in phase I trials of anticancer agents, attempts to decrease the number of patients per dose level and to conduct a larger dose escalation have been made. Among them, accelerated titration designs were proposed and evaluated by simulation; designs 2 and 4 were reported to be acceptable (J Natl Cancer Inst 89: 1138-1147, 1997). Both designs 2 and 4 included only one patient per cohort during the initial accelerated phase. Dosage steps for the accelerated phase were defined using the modified Fibonacci method for design 2 and 100% escalation for design 4, respectively. The accelerated phase continued until one patient experienced dose-limiting toxicity or two patients experienced grade 2 toxicities. Dose escalation was conducted based on the information from the first course in design 2 and from the first three courses in design 4. In the simulation, both designs successfully reduced the total number of patients and the number of undertreated patients without increasing the number of overtreated patients. However, the safety of design 4 was assured as long as all patients received three courses of chemotherapy, which is unusual in phase I studies in Japan. Decision-making on dose escalation based on the information on toxicity in three courses might be cumbersome. Therefore, in Japan, design 2 would be recommended among the proposed accelerated designs. The performance of the design should be investigated by applying it to actual phase I studies and by evaluating the number of undertreated and overtreated patients. PMID- 11016011 TI - Thrombolytic therapy for acute myocardial infarction: what dose should be used? AB - Most of the concern about dosing of t-PA as presented in the literature is related to the lower weight groups and the increased incidence of bleeding in this patient population. Not much is known or written about what doses of t-PA should be used in the grossly obese patient with an MI. In the patient described briefly at the beginning of this editorial, the usual accelerated dose regimen was used. Although the patient's symptoms resolved quickly, ST-segment elevation never returned to baseline. Creatine kinase-MB and troponin I rose and fell slowly. He had an uneventful course in the hospital but my impression is that he did not reperfuse his myocardium. Unfortunately, because of his weight, we were unable to evaluate this patient with either coronary angiography or echocardiography. As I think more about this case, I am of the opinion that the patient was underdosed with 100 mg of t-PA. If I had it to do over again, I would use a higher dose of t-PA. I would be interested to hear how others would have treated the patient. PMID- 11016012 TI - Pulmonary embolus after coronary artery bypass surgery: a review of the literature. AB - Pulmonary embolus (PE) after cardiac bypass surgery is an uncommon complication but carries with it high morbidity and mortality. The incidence of deep vein thrombosis (DVT) and PE after cardiac bypass varies depending on postoperative thromboprophylaxis, the presence of indwelling central venous catheters in the lower extremities, and early ambulation. The clinical diagnosis of DVT remains difficult and challenging. Pulmonary embolus is often the first occurring clinical event. The safety and effectiveness of preventative pharmacologic agents, such as subcutaneous unfractionated or fractionated heparin or oral coumadin, remain largely unknown. Heparin-induced thrombocytopenia, generally associated with a high incidence of DVT and PE, occurs in approximately 3.8% of patients who have undergone cardiac surgery and are placed postoperatively on high-dose intravenous unfractionated heparin. Sequential compression devices (SCD) have not been effective in reducing the incidence of DVT in an ambulating cardiac bypass patient when added to routine elastic graded compression stockings (GCS). Very large clinical trials are necessary to prove the effectiveness of pharmacologic or mechanical preventative measures in reducing the incidence of PE after cardiac surgery above the commonly used GCS, early ambulation, and aspirin. In a nonambulating, higher-risk cardiac bypass patient with slow recovery, a more aggressive prophylaxis regimen might be necessary for optimal prevention, although further data are needed to support this hypothesis. PMID- 11016013 TI - Endothelial dysfunction, subangiographic atheroma, and unstable symptoms in patients with chest pain and normal coronary arteriograms. AB - BACKGROUND: Patients with chest pain and normal coronary arteriograms (CPNA) may present with unstable symptoms and other evidence of ischemia during clinical follow-up. Although repeat angiography usually proves negative, functional assessment of coronary vasomotor abnormalities may provide additional pathophysiologic information. HYPOTHESIS: The study was undertaken to evaluate the relationship between endothelial dysfunction and subangiographic atheroma in patients with CPNA undergoing repeat angiography because of unstable symptoms. METHODS: We investigated nine patients with CPNA (8 women, mean age 57 +/- 9 years) undergoing repeat angiography because of unstable anginal symptoms. After normal angiography, simultaneous coronary epicardial and microvascular vasomotor responses to intracoronary vasodilators [acetylcholine (10(-6) M), adenosine (18 micrograms) and nitroglycerin (300 micrograms)] were investigated in the left anterior descending artery using quantitative angiography and Doppler flow measurements. The presence of subangiographic atheroma was assessed by intravascular ultrasound. RESULTS: Three patients demonstrated proximal and distal epicardial vasoconstriction and a reduction in coronary flow in response to acetylcholine, indicating concordant epicardial and microvascular endothelial dysfunction. These changes were associated with chest pain and ischemic electrocardiographic changes in two patients. None of the remaining patients suffered chest pain in response to intracoronary acetylcholine. Six patients had significant subangiographic disease (intimal thickness > 0.3 mm) on intravascular ultrasound imaging, and multivariate analysis indicated a significant relationship (R2 = 0.89, overall p = 0.001) between the extent of subangiographic disease and both plasma cholesterol concentration and hypertensive history. No significant relationship was demonstrated between endothelial dysfunction and the extent of subangiographic disease. CONCLUSION: Concordant epicardial and microvascular endothelial dysfunction may be pathophysiologically and clinically significant in unstable patients with CPNA but does not appear to be directly related to the extent of subangiographic atheroma. PMID- 11016014 TI - Susceptibility of apolipoprotein B-containing lipoproteins to oxidation and antioxidant status in acute coronary syndromes. AB - BACKGROUND: Oxidized lipoproteins may play an important role in the pathogenesis of atherosclerosis, and it has been shown that antioxidants have a protective effect against the progression of atherosclerosis. HYPOTHESIS: The aim of this study was to investigate the oxidative susceptibility of apolipoprotein B containing lipoproteins and antioxidant status in patients with acute coronary syndromes and chronic stable angina pectoris. METHODS: The study population included 70 patients with acute coronary syndromes (14 with recent acute myocardial infarction and 56 with unstable angina pectoris), 105 patients with stable angina pectoris, and 75 control subjects. In addition to conventional lipid and lipoprotein analysis, the susceptibility of apolipoprotein B-containing lipoproteins to in vitro oxidation (lag phase) and plasma vitamin E and total carotene levels was measured. RESULTS: The lag phase was significantly shorter in patients with acute coronary syndromes (45 +/- 12 min) than in patients with stable angina pectoris (51 +/- 10 min) and in control subjects (58 +/- 9 min) (p < 0.0001). Both plasma vitamin E and total carotene levels were lowest in patients with acute coronary syndromes (1.11 +/- 0.32 mg/dl and 119 +/- 32 micrograms/dl, respectively), followed by patients with stable angina pectoris (1.25 +/- 0.37 mg/dl and 132 +/- 37 micrograms/dl) and then controls (1.52 +/- 0.31 mg/dl and 167 +/- 41 micrograms/dl). CONCLUSIONS: These data suggest that there is an intense oxidative process and a lower antioxidant status in acute coronary syndromes. This may lead to plaque instability due to the activation of the inflammatory response in coronary atherosclerotic lesions. PMID- 11016015 TI - Worsening of left ventricular diastolic dysfunction during exercise causes decreased exercise tolerance in hypertension. AB - BACKGROUND: Exercise tolerance is reduced in hypertension. Hypertension affects left ventricular (LV) diastolic filling by causing abnormal relaxation and decreasing compliance. HYPOTHESIS: This study was designed to determine whether worsening of LV diastolic dysfunction during exercise causes decreased exercise tolerance in hypertension. METHODS: Left ventricular diastolic filling parameters were examined at mitral valve by Doppler echocardiography at rest and at peak exercise in hypertensive patients and were compared with those of age- and gender matched normotensive individuals. Treadmill exercise stress test was performed according to the Bruce protocol and the exercise time was recorded. RESULTS: Exercise time was significantly shorter in the hypertensive group than that in the normotensive group (320 +/- 29 vs. 446 +/- 38 s, p 0.03). The hypertensive group demonstrated abnormal relaxation pattern of diastolic mitral inflow at rest, which became pseudonormal at peak exercise (E/A velocity ratio, rest 0.86 +/- 0.06 vs. exercise 1.19 +/- 0.09, p < 0.001). The diastolic mitral inflow pattern remained normal at peak exercise in the normotensive group. The deceleration time and the pressure half time of early mitral inflow at peak exercise were significantly shorter in the hypertensive group than those in the normotensive group (deceleration time, 182 +/- 20 vs. 238 +/- 22 ms, p 0.02: pressure half time, 54 +/- 5 vs. 70 +/- 12 ms, p 0.01). CONCLUSIONS: This study demonstrates that reduced exercise tolerance in hypertension is associated with worsening of diastolic dysfunction during exercise consistent with an increase in left atrial pressure. PMID- 11016016 TI - Diastolic ventricular interaction in normal and dilated heart during head-up tilting. AB - BACKGROUND: The normal human heart behaves as a single functional unit during preload reduction; adaptations of the left ventricle to head-up tilting is mediated through ventricular interdependence and biventricular-lung interaction. HYPOTHESIS: We hypothesized that reduction of venous return in dilated cardiomyopathy is likely to have a great effect on ventricular chamber geometry and filling. The aim of this study was to evaluate the effects of gradual head-up tilting in normal subjects and in patients with dilated cardiomyopathy, addressing special attention to right (RV) and left ventricular (LV) dimensions, geometry, and filling, and to biventricular-lung interaction. METHODS: Twenty normal subjects and 23 patients with moderate heart failure due to dilated cardiomyopathy were studied with two-dimensional and Doppler echocardiography in supine position and after 20 degrees, 40 degrees, and 60 degrees tilting. Right ventricular and LV dimensions, LV geometry, and tricuspid, mitral, and pulmonary venous flow patterns were recorded at each step of the study. Geometric changes of the LV were evaluated by measurements of volumes and diameters in the apical four-chamber view (which identifies the interventricular septum and lateral wall) and apical two-chamber view (which identifies the inferior and anterior wall of the LV). RESULTS: In the two groups, tilting was associated with reduction of RV area and LV diameter and volumes; percent variations in LV diameter and volumes recorded in four-chamber view were lower at each step of tilting than with those derived from the two-chamber view in controls and in patients. In normal subjects, mitral and tricuspid peak early flow velocities were decreased at any tilting level; peak late velocities were unchanged; peak velocity of systolic forward flow of the pulmonary vein was reduced, diastolic forward flow was unchanged, and the difference in duration between reverse pulmonary flow and forward mitral A wave was reduced. Doppler findings were qualitatively similar in patients, but tilting induced a more marked redistribution of LV filling to late diastole because of a significant increase in atrial contribution. CONCLUSIONS: Preload reduction by tilting induces profound effects on left and right dimensions, geometry, and filling in normal and dilated heart; reduction or RV dimensions are associated with changes in LV ventricular geometry (minimal reduction in septal-lateral diameter, marked reduction in anterior-posterior diameter), redistribution of right and left diastolic filling to late diastole, and redistribution of pulmonary venous flow to early diastole. These mechanisms are probably due to a favorable interaction between heart and lungs, which increases compliance within the pericardial space and facilitates redistribution of flow from the lungs. Even a minimal amount of preload reduction causes more marked effects in LV filling patterns in dilated cardiomyopathy than in normal hearts, confirming that ventricular interaction and pericardial constraint are increased when heart volume enlarges. PMID- 11016017 TI - Hemodynamic effects and long-term outcome of percutaneous balloon valvuloplasty in patients with mitral stenosis and atrial fibrillation. AB - BACKGROUND: The presence of atrial fibrillation (AF) has been identified as a predictor of a suboptimal result in some patients undergoing percutaneous balloon valvuloplasty in the treatment of symptomatic rheumatic mitral stenosis. HYPOTHESIS: Atrial fibrillation adversely affects the short- and long-term outcome of patients with mitral stenosis undergoing percutaneous balloon valvuloplasty. METHODS: A retrospective chart review of 104 consecutive patients with rheumatic mitral stenosis undergoing percutaneous balloon valvuloplasty was performed. A successful procedure was defined as a final mitral valve area > or = 1.5 cm2 and the absence of a complication. Endpoints included freedom from mitral valve replacement, death, and repeat balloon valvuloplasty at 5 years. RESULTS: A successful procedure was obtained in 89% of patients with sinus rhythm and in 78% of patients with AF (p = NS). Patients in sinus rhythm had a greater cardiac output resulting in a larger final valve area than patients in AF (1.8 vs. 1.6 cm2, p < 0.05). Freedom from valve replacement, death, and repeat balloon valvuloplasty at 5 years was 75% for patients in AF and 76% for patients in sinus rhythm (p = NS). Lower postprocedure mitral regurgitation grade and absence of prior commissurotomy were the only independent predictors of event-free survival. CONCLUSIONS: Patients with mitral stenosis and AF have lower cardiac outputs and gradients than patients with sinus rhythm, despite similar valve areas. The long term outcome of balloon valvuloplasty is independent of the initial cardiac rhythm. PMID- 11016018 TI - DDD and single-lead VDD pacing: evaluation of atrial signal dynamic changes. AB - BACKGROUND: Single-lead VDD pacing systems are an alternative to conventional DDD pacemakers in patients with atrioventricular (AV) block and normal sinus function. HYPOTHESIS: The aim of this study was to assess changes of P-wave amplitude occurring in dynamic conditions in two groups of patients with a single lead VDD and with a DDD pacing system, respectively. METHODS: Twenty-eight patients with second- or third-degree AV block and normal sinus function were enrolled prospectively into the study. Seventeen patients were implanted with a single-lead VDD pacing system and 11 with a DDD pacemaker. Patients were evaluated at 3 months (all patients) and at 6 months (26 patients) at supine and in dynamic conditions (postural changes, hyperventilation, and during exercise). RESULTS: Mean P-wave values at supine were 1.92 +/- 1.10 mV at 3 months and 1.76 +/- 1.01 mV at 6 months for VDD systems, and 4.63 +/- 2.18 mV at 3 months and 4.58 +/- 2.80 mV at 6 months for DDD pacemakers. In dynamic conditions, P-wave amplitude changes compared with supine condition ranged between -74 and +226% in VDD, and between -53 and +138% in DDD; however P-wave amplitudes showed no significant changes compared with baseline. Moreover, changes in atrial signal amplitudes did not occur randomly, and in both systems P-wave amplitudes remained significantly correlated with supine values. CONCLUSIONS: A wide range of P-wave amplitude variations occurs in different postural conditions or during exercise, both with single-lead VDD and DDD pacing systems. However, with appropriate programming of atrial sensitivity based on supine values, constant atrial tracking can be maintained. PMID- 11016019 TI - Comparison of the efficacy and tolerability of simvastatin and atorvastatin in the treatment of hypercholesterolemia. AB - BACKGROUND: Simvastatin and atorvastatin are effective statins for treating hypercholesterolemia. HYPOTHESIS: The study was undertaken to compare the efficacy and tolerability of simvastatin 20 and 40 mg/day and atorvastatin 10 and 20 mg/day. METHODS: In this multinational, open-label, crossover study, 258 patients with primary hypercholesterolemia were randomized after 4 weeks of diet plus placebo to once-daily administration of a starting dose sequence of simvastatin (20 mg) or atorvastatin (10 mg), or a higher dose sequence of simvastatin (40 mg) or atorvastatin (20 mg) for 6 weeks. Patients were then switched after a 1-week washout to the corresponding starting or higher dose of the alternate drug for a second 6-week period. The primary endpoint was the mean percent change from baseline to Week 6 in low-density lipoprotein (LDL) cholesterol; percent changes from baseline in total cholesterol, high-density lipoprotein cholesterol, triglycerides, and apolipoprotein B were also compared. Safety was assessed through adverse experiences and laboratory measurements. RESULTS: Both statins produced statistically significant improvements in all measured plasma lipids and lipoproteins. The main treatment comparison showed no statistically significant difference in changes in LDL cholesterol and triglycerides, whereby the overall effects were comparable when doses of 20 mg and 40 mg of simvastatin were compared with atorvastatin 10 mg and 20 mg. The mean percent reductions for LDL cholesterol from baseline to Week 6 ranged from 35-42% for the entire study cohort. An LDL cholesterol level < or = 130 mg/dl (3.4 mmol/l) was achieved in approximately 70% of patients treated with both drugs in this study. Simvastatin and atorvastatin were well tolerated at the doses studied. CONCLUSION: In patients with hypercholesterolemia, the most commonly used doses of simvastatin and atorvastatin produced similar changes in LDL cholesterol and achieved an LDL cholesterol level < or = 130 mg/dl (3.4 mmol/l) in a similar number of patients. Both statins were well tolerated. PMID- 11016020 TI - Effect of cibenzoline on regional left ventricular function in hypertrophic obstructive cardiomyopathy. AB - BACKGROUND: Cibenzoline, a class Ia antiarrhythmic drug, can be used to relieve left ventricular (LV) outflow obstruction in hypertrophic obstructive cardiomyopathy (HOCM). However, the mechanism of this agent in HOCM has been controversial. HYPOTHESIS: This study was designed to investigate the effect of cibenzoline on regional LV function and the acoustic properties in HOCM using ultrasonic integrated backscatter. METHODS: Ten patients with HOCM and 16 healthy volunteers were examined. In patients with HOCM, wall thickening (%WT) and the magnitude of cyclic variation of integrated backscatter (mag-CVIBS) in the interventricular septum (IVS) and LV posterior wall were measured before and after oral administration of cibenzoline. To assess asynchrony of contractile elements, the phase difference between CVIBS and %WT were measured from the LV posterior wall. Pressure gradients at the LV outflow tract were estimated using continuous-wave Doppler echocardiography. RESULTS: Although %WT decreased significantly in the LV posterior wall, %WT and mag-CVIBS remained unchanged in the IVS. The phase difference in the LV posterior wall was significantly greater in patients with HOCM than in healthy volunteers (HOCM:healthy volunteers, 1.57 +/- 0.23:1.00 +/- 0.03, p < 0.001) at baseline. After administration of cibenzoline, the phase difference shifted to normal value (from 1.57 +/- 0.23 to 1.28 +/- 0.27, p = 0.0382), and pressure gradients at the LV outflow tract decreased (from 109 +/- 55 to 58 +/- 48 mmHg, p = 0.0063). Changes in pressure gradients at the LV outflow tract and the phase difference were closely related. CONCLUSIONS: Regional function and the acoustic properties of myocardium in HOCM were altered by cibenzoline in the LV posterior wall but remained unchanged in the IVS. The normalization of the phase difference in the LV posterior wall was closely related to the decrease in pressure gradients at the LV outflow tract. These findings suggest that negative inotropic action and the improvement of asynchrony in the LV posterior wall rather than in the IVS may contribute to the reduction of pressure gradients at the LV outflow tract in HOCM. PMID- 11016021 TI - Fast platelet suppression by lysine acetylsalicylate in chronic stable coronary patients. Potential clinical impact over regular aspirin for coronary syndromes. AB - BACKGROUND: The rapid utilization of fibrinolytics following Q-wave myocardial infarction has clearly modified the evolution of this disease. However, it is still not known whether the immediate inhibition of platelet aggregation (PA) during the coronary event improves outcomes. HYPOTHESIS: The present study was designed to test, in patients with known coronary artery disease (chronic stable angina), whether the particular kinetic pattern of lysine acetylsalicylate (LA) compared with aspirin may affect the time to onset of inhibition of platelet aggregation. METHODS: Ten patients suffering from chronic stable angina participated in this study to compare the efficacy and speed of the inhibition of PA with 320 mg of LA versus 320 mg of aspirin. All patients discontinued the use of aspirin and any other anti-inflammatory agents for 15 days prior to the beginning of the study. They were randomly assigned to LA or aspirin. Blood specimens were obtained to measure the PA at admission, and 5, 10, 20, 30, and 60 min after ingestion. Patients continued to take the assigned drug once a day for the following 4 days. On Day 5, a new blood sample was taken. After this, patients underwent a 15-day wash-out period, and then crossed over to the opposite drug. The samples were analyzed immediately using platelet-rich plasma stimulated with adenosine diphosphate (ADP) 2 mumol/l, collagen 1 microgram/ml, epinephrine 20 mumol/l, and sodium arachidonate acid 0.75 mm/l. RESULTS: The same level of PA inhibition after 30 and 60 min of aspirin administration can be obtained with LA 5 min following ingestion (sodium arachidonate acid: LA: 16.3 +/ 25.9 vs. aspirin 57.6 +/- 8.2; p = 0.00014; collagen: LA 18.9 +/- 20.1 vs. aspirin 47.2 +/- 10.5; p = 0.00092; ADP: LA 27.3 +/- 18.4 vs. aspirin 39.7 +/- 21.8, p = 0.18; epinephrine: LA 22.0 +/- 9.9 vs. aspirin 55.4 +/- 10.9, p = 0.00002. CONCLUSIONS: Platelet aggregation inhibition immediately following LA may have significant clinical implications for the treatment of coronary syndromes. PMID- 11016022 TI - Tissue Doppler features of cardiac amyloidosis. PMID- 11016023 TI - Intracardiac masses detected by echocardiography: case presentations and review of the literature. AB - Intracardiac masses are often diagnosed by transthoracic echocardiography (TTE). Transesophageal echocardiography (TEE) improves overall visualization of masses, especially those located in the posterior cardiac structures. Masses in the heart are most commonly due to thrombi or valvular vegetations; however, a variety of tumors may also present as cardiac masses on echocardiography. Tumors of the heart most commonly occur in the setting of metastatic disease, usually from malignancies of the breast, lung, or from malignant melanoma. Primary cardiac tumors occur much less frequently and are usually benign. Atrial myxomas constitute nearly one-half of reported primary cardiac tumors. The following discussion details the findings of five cases that illustrate the spectrum of intracardiac tumors detected by echocardiography and reviews the relevant literature. PMID- 11016024 TI - Profiles in cardiology. Ernst, Wilhelm, and Eduard Weber. PMID- 11016026 TI - Current status and future technical advances of ultrasonic imaging. PMID- 11016027 TI - Advances in nuclear emission PET and SPECT imaging. PMID- 11016028 TI - Current technical development of magnetic resonance imaging. PMID- 11016030 TI - Challenges of imaging structure and function with MRI. PMID- 11016029 TI - Technical challenges of functional magnetic resonance imaging. PMID- 11016031 TI - System performance of multislice spiral computed tomography. AB - Multislice spiral CT offers many new possibilities for clinical CT imaging. Drastically increased scan speeds and z-resolution, respectively, as well as applications such as cardiac CT that have become feasible for the first time in routine clinical use. The concept of multiple simultaneously acquired slices yields image quality equivalent or better than single-slice spiral CT. Especially, there is no dependence on spiral pitch, neither with regard to noise nor to slice sensitivity. The reconstructed slice width can be chosen freely and retrospectively, which offers additional flexibility when evaluating optimal protocols for various kinds of examinations. Three-dimensional isotropic resolution can be achieved routinely with examinations fast enough to scan in a single breath hold (Fig. 9). Without any drawbacks in image quality, MSCT in combination with online tube current modulation can reduce patient dose. In some body regions, dose is decreased to 50% compared to a scan with constant tube current. One of the most promising new applications is the dedicated ECG-gated cardiac interpolation 180 degrees MCI, which allows four-dimensional (4-D) imaging of the heart. The complete beating heart can be reconstructed in well defined phases of the heart cycle, thereby adding high temporal resolution to isotropic 3-D spatial resolution (for more examples refer to http://www.imp.uni erlangen.de/e/research/cardio/). PMID- 11016032 TI - Imaging the lungs with computed tomography. PMID- 11016033 TI - Current status and future advances of digital radiography and PACS. PMID- 11016034 TI - Computerized decision support in medical imaging. PMID- 11016035 TI - Automated keratometry at low cost. PMID- 11016037 TI - Nonlinear dynamic characteristics analysis of synchronous 12-lead ECG signals. PMID- 11016036 TI - Evaluating arrhythmias in ECG signals using wavelet transforms. PMID- 11016038 TI - Horatio B. Williams and the first electrocardiographs made in the United States. PMID- 11016040 TI - Cost-effectiveness analysis: an essential tool for practice management. PMID- 11016039 TI - Tissue engineering applied to reconstructive surgery. PMID- 11016041 TI - Assessing ambulatory geriatric sleep complaints. AB - Sleep disturbances afflict more than 50% of adults age 65 and older. Sleep apnea and periodic limb movements of sleep are the primary sleep disorders in the elderly. Patient assessment tools, a thorough physical examination, and appropriate tests can simplify the diagnostic process; sleep center referral is not always warranted. Ultimately, accurate sleep disorder diagnoses can result in decreased geriatric morbidity and mortality, and increased patient quality of life. PMID- 11016042 TI - Managing sinusitis in children. AB - Because of the prevalence of upper respiratory tract infections (URIs), sinusitis is a condition commonly encountered in the pediatric population. Some 5% to 10% of children with URIs also have sinusitis. A thorough clinical evaluation enables the health care provider to accurately diagnose sinusitis in children without overuse of computed tomography scans or antibiotics. This article discusses the diagnosis and treatment of acute sinusitis in children. PMID- 11016043 TI - Herbal and nutritional supplement use in the elderly. AB - Herbal and nutritional supplement use in the United States is increasing, including use among older adults. Often, older adults do not inform health care providers that they are using dietary supplements. Dietary supplement use can be problematic because of potential interactions with prescription drugs, patient delays in seeking needed care, and inadequate product quality control. This article provides guidelines for dietary supplement use in the older adult and discusses the precautions associated with common herbs. PMID- 11016044 TI - Glucosamine and osteoarthritis. PMID- 11016045 TI - What is happening to children with failure to thrive? PMID- 11016046 TI - Approaches to the diagnosis and management of growth failure. PMID- 11016047 TI - The dilemma of the short child without a clear diagnosis. PMID- 11016048 TI - A clinical genetics and dysmorphology approach to growth deficiency. PMID- 11016049 TI - A gastroenterologist's approach to failure to thrive. PMID- 11016050 TI - The basics for the diagnosis and management of short stature: a pediatric endocrinologist's approach. PMID- 11016052 TI - A parent's view of short stature. PMID- 11016051 TI - Endocrine treatments for short stature. PMID- 11016053 TI - Resident's column: "Doctor, is my child growing ok?". PMID- 11016054 TI - [Sex education of children and adolescent]. AB - There is a remarkable ignorance about sexuality among Chilean teenagers, as underscored by the paper by Fernandez et al, published in this issue. The authors analyze the biological, psychological and social consequences of the lack of parental and school sexual education. Among them, there is an increasing incidence of unwanted pregnancies, abortions and sexually transmitted diseases in teenagers. Child abuse, neglect, battering, mortality and maternal deprivation are more frequent among the offspring of teenage parents. School desertion caused by unwanted pregnancies will seriously jeopardize the education and training of these teenagers, perpetuating the cycle of poverty. PMID- 11016055 TI - [Beliefs, attitudes and knowledge about sex education]. AB - BACKGROUND: Previous reports show that Chilean teenagers have an inadequate knowledge about sexuality and reproduction. AIM: To compare the knowledge about sexuality among adolescents coming from private and public schools, with and without sexual education programs. MATERIAL AND METHODS: A structured anonymous inquiry, containing multiple choice and open questions, was applied to a sample of 229 adolescents attending seventh and eighth grade of junior school, in private and public schools of Temuco, Chile. RESULTS: Eleven percent of adolescents had already their first sexual intercourse at a mean age of 12.2 +/- 2.4 years old. Of these, 96% came from public schools. An overall analysis of tests, disclosed a 53% of correct answers to the inquiry. Adolescents coming from private schools had a better performance than those coming from public schools. Sexual attitudes were not influenced by sexual education programs. CONCLUSIONS: Adolescents coming from private schools have a better sexual knowledge level and more conservative attitudes towards sexuality. Overall knowledge is inadequate albeit overvalued. These teenagers are high risk group for unwanted pregnancies and sexually transmitted diseases and require efficient sexual education programs. PMID- 11016056 TI - [Increased activity of metalloproteinases and their inhibitors in cerebrospinal fluid of patients with tropical spastic paraparesis]. AB - BACKGROUND: Proteolytic modifications of neuronal surfaces and the surrounding extracellular matrix are very important in neuronal development and regeneration. Increased activity of matrix metalloproteinases (MMPs) and their tissue inhibitors, due to secretion by macrophages and lymphocytes, occur in inflammatory processes that disrupt the blood brain barrier. However, neurons and microglia can also secrete these enzymes. AIM: To identify the type of MMP present in the cerebrospinal fluid (CSF) and changes in the expression of tissue inhibitors of metalloproteinases (TIMPs) in patients with HTLV-1 associated tropical spastic paraparesis. PATIENTS AND METHODS: CSF samples from 12 patients with HTLV-1 associated tropical spastic paraparesis and 12 healthy controls were obtained by an atraumatic lumbar puncture. The presence of MMPs was measured by zymography and the relative amounts of TIMPs were measured by immunowestern blot. RESULTS: In the CSF of both controls and patients, a similar gelatinolytic band corresponding to proMMP-2 (latent form) was observed. In 83.3% of patients with HTLV 1 associated tropical spastic paraparesis, the MMP-9 was also present. TIMP 1, TIMP-2 and TIMP-3 were elevated 2.24 +/- 0.72, 3.85 +/- 1.38 and 5.89 +/- 3.4 fold, respectively, in the CSF of patients as compared to controls. CONCLUSIONS: Patients with HTLV-1 associated tropical spastic paraparesis have elevated activity of MMP-9 and levels of TIMPs in the CSF, when compared to healthy controls. PMID- 11016057 TI - [Genetic composition of Chilean population: rural communities of Elqui, Limari and Choapa valleys]. AB - BACKGROUND: The population that inhabits the semiarid Northern zone of Chile arose from ethnic admixture between aborigines, Spanish conquerors and the influx, during the XVII century, of foreign aboriginal workers and a minority of African slaves. AIM: To study the phenotypic frequencies of 15 genetic markers among populations inhabiting valleys in the Northern zone of Chile and to estimate the percentage of indigenous, African and Caucasian admixture in these populations. MATERIAL AND METHODS: Throughout five different field works, blood samples were obtained from 120 individuals living in the Elqui valley, 120 individuals living in the Limari valley and 85 living in the Choapa valley. Blood groups, erythrocyte enzymes, plasma proteins and HLA markers were typified. RESULTS: In the populations studied, the contribution of non indigenous genes was low in relation with the time elapsed since the Spanish invasion. The Hardy Weinberg disequilibrium for MNS system would have microevolutive implications. The admixture percentages in these valleys confirm ethnic and historic information. The variation of the enzyme esterase D is identical to that of other Chilean populations. CONCLUSIONS: The phenotypic and genetic frequencies in the three populations studied and different admixture of indigenous genes is inversely proportional to the geographic distance from Santiago, in Central Chile. PMID- 11016058 TI - [Primary epiploic appendicitis: clinical and radiological diagnosis]. AB - BACKGROUND: Torsion of appendices epiplocae leads to an ischemic infarct of surrounding adipose tissue, causing a syndrome, called "primary epiploic appendagitis" characterized by acute abdominal pain that can simulate a surgical clinical picture. AIM: To describe the clinical picture, ultrasonographic and computed tomographic features of primary epiploic appendagitis. MATERIAL AND METHODS: A report of patients with acute abdominal pain whose diagnosis was a primary epiploic appendagitis, diagnosed and treated in a period of 48 months. RESULTS: Fifty five patients (45 males) aged 16 to 76 years old are reported. Their clinical presentation was acute abdominal pain in the left abdominal quadrant in 48, pain in the right lower quadrant in 4 and epigastric pain in two. Two had mild fever and 12 had nausea. Ten perform physical activities prior to the onset of pain. Imaging examinations showed a 1.5 to 5 cm diameter, uncompressible small mass of adipose origin, located anteriorly and anterolaterally, between the colon and the abdominal wall. There were inflammatory phenomena surrounding the lesion and thickening of the neighboring parietal peritoneum. In all cases, the mass gradually subsided with medical treatment. CONCLUSIONS: Primary epiploic appendagitis is a relatively frequent cause of spontaneously resolving abdominal pain. It is diagnosed by ultrasound or CT scanning. PMID- 11016059 TI - [Treatment of diffuse hyperthyroid goiter with radioiodine: influence of propylthiouracil pretreatment]. AB - BACKGROUND: To stabilize Graves disease and deplete the preformed hormone, the use of antithyroid drugs prior 131I therapy has been suggested, specially in those patients with severe thyrotoxicosis and in the elderly. However, PTU may reduce the effectiveness of 131I. AIM: To study the effects of PTU pretreatment before 131I administration. SUBJETS AND METHODS: A retrospective analysis of the medical records of patients with Graves disease treated with 131I from 1989 to 1997 was made. Of 244 patients with adequate follow-up for at least 12 months after 131I treatment, 142 had not been pretreated and 102 had received PTU prior to 131I therapy. Pretreated patients were distributed according to the number of days that PTU was discontinued before receiving 131I, forming four groups (a = 5 d, b = 6-14 d, c = 15-30 d and d = 31-60 d). Radioiodine was delivered according to our protocol of 120 microCi per gram of thyroid tissue, as estimated by clinical examination. Therapy was considered successful when laboratory evidence of euthyroidism or hypothyroidism after one year of treatment was obtained and as a failure when undetectable TSH values persisted after 12 months of treatment with 131I. RESULTS: All groups were comparable as to age, gender, goiter size, and 24 h radioiodine uptake. Control of hyperthyroidism was achieved in 76% of the non pretreated group. A similar percentage was observed in groups (b), (c) and (d). However, the disease was controlled in only 50% of group (a) patients (p < 0.003). CONCLUSIONS: The therapeutic efficacy of 131I is significantly reduced when the PTU is stopped for only a few days prior to the use of radioiodine. We postulate that PTU has to be discontinued for at least 10 days before radioiodine administration. PMID- 11016060 TI - [Cross cultural psychometric comparison of major depression in Chile and Germany]. AB - BACKGROUND: Depressive symptoms and personality features of patients with mood disorders are supposed to have a strong variability in cross cultural studies. The clinical profile, the outcome and the treatment response seem to be different across the world. AIM: To investigate the differences and similarities of major depressive disorders in Chile and Germany. MATERIAL AND METHODS: Sixty seven Chilean and German depressive inpatients, without comorbidity on axis I or II were studied. Diagnosis of depression was based on DSM IV and ICD 10 criteria for major depression. Symptomatology was assessed using the self reporting questionnaire (SRQ-20), the Symptom checklist-90-revisited (SCL 90-R), the Beck inventory for Depression (BDI), and the Hamilton scale for depression (HAM-D). Personality was assessed with the Munich Personality Scale. RESULTS: Only the SRQ 20 shows significantly higher tendency towards depression among Chilean patients. At the personality level, Chileans exhibited higher scores in extroversion, neuroticism, esoteric and isolating tendencies. CONCLUSIONS: These results help to understand differences and similarities between depressive syndromes across cultures. They suggest that the relation between symptomatology, some personality traits and severe major depression has little variability across cultures. PMID- 11016061 TI - [Canalith reposition procedure for the treatment of benign paroxysmal postural vertigo]. AB - BACKGROUND: Paroxysmal positional vertigo is a frequent and handicapping disease. AIM: To assess the effectiveness of physical therapies using particle reposition procedures in the treatment of paroxysmal positional vertigo. PATIENTS AND METHODS: Fifty nine patients (13 male) with paroxysmal positional vertigo were studied. Clinical data was gathered, the affected channel was identified. In the same session, the corresponding canalith reposition procedure was performed (Epley maneuver for posterior channel and Lempert maneuver for lateral channel) and the immediate response was recorded. RESULTS: Twenty seven percent of patients had recurring episodes of vertigo and 60% had more than two weeks of evolution. In 49% of patients, vertigo was considered idiopathic and in 92%, the posterior channel was affected. One patient had a combined lesion of posterior and lateral channels and two patients had isolated lateral channel lesions. Seventy percent of patients recovered after two sessions of canalith reposition procedure. CONCLUSIONS: Canalith reposition maneuvers were highly effective for the treatment of paroxysmal positional vertigo. PMID- 11016062 TI - [Risk factors for home deaths due to pneumonia among low socioeconomic level Chilean children, Santiago de Chile (1994)]. AB - BACKGROUND: Pneumonia is the main cause of late infant mortality in Chile. Over 60% of these deaths occur at home. The lack of hospital beds and the inadequate outpatient management are contributing factors. AIM: To assess risk factors for home deaths due to pneumonia in Chilean children. PATIENTS AND METHODS: The clinical and environmental histories of 53 (39 male) children that died due to pneumonia in their homes were analyzed. The cause of death was confirmed by necropsy with histopathological studies in all cases. These cases were compared with 88 control children of similar age, gender, socioeconomic status and living in the same geographical area of Metropolitan Santiago. RESULTS: Fifty four percent of deceased children were of less than 3 months of age and only 3 cases and their controls were above 1 year old. Identified risk factors for death were malnutrition with an odds ratio of 30.6 (CI 3.9-64.8, p < 0.001), low birth weight with an odds ratio of 5 (CI 1.8-14.1, p < 0.001), previous admissions to hospitals with an odds ratio of 5.79 (CI 2-17.1, p < 0.001), congenital malformations (mainly cardiac) with an odds ratio of 8.4 (CI 2-39.9, p = 0.001) and a history of bronchial obstruction with an odds ratio of 5.68 (p < 0.001). Identified maternal risk factors were smoking with an odds ratio of 4.13 (CI 1.6 10.7 p < 0.001) and being a teenager with an odds ratio of 4.3 (CI 1.7-11, p < 0.001). Malnutrition, low birth weight, history of previous hospital admissions and having a teenager or smoker mother were considered as independent risk factors using a stepwise analysis. CONCLUSIONS: Chilean low income children have identifiable risk factors for death at their homes due to pneumonia, that can be preventively managed. PMID- 11016063 TI - [Bronchiolitis obliterans with organizing pneumonia secondary to chemotherapy in a child with primary pericardial sarcoma]. AB - The uncommon bronchiolitis obliterans organizing pneumonia can be idiopathic or caused by infection or medications. We report a 5 year old boy with pericardial sarcoma that was treated with chemotherapy (vincristine, doxorubicin, etoposide, cyclophosphamide) and radiotherapy. During the third cycle of chemotherapy, he developed progressive cough and dyspnea, needing oxygen therapy. Chest X-ray examination showed bilateral infiltrates. After discarding infectious etiologies, an open lung biopsy was performed, and the pathological examination showed a bronchiolitis obliterans organizing pneumonia. He was treated with prednisolone for 7 days, followed by prednisone for additional 45 days. He continued with fluticasone propionate as maintenance and remains in good condition with a chest X-ray showing mild interstitial images, after seven months of follow up. PMID- 11016064 TI - [Invasive hepatosplenic hyalohyphomycosis. Report of a case]. AB - Invasive fungal infections are an increasingly common problem in patients with cancer and other vulnerable groups. We report a case of hepatosplenic hialohifomycosis probably by Aspergillus sp. In a patient with acute leukemia, and prolonged neutropenia, treated with corticosteroids and broad spectrum antibiotics. A review concerning diagnosis, clinical course and treatment of this condition is presented. PMID- 11016065 TI - [Hemorrhagic syndrome caused by exposure to a rodenticide. Report of a case]. AB - We report a 22 years old male, admitted to the emergency room due to a life threatening coagulation disorder, with prothrombin times fluctuation between 5 and 37% and very low activity of factors II, VII, IX and X. In the month prior to the admission, the patient had used the rodenticide difethialone, without any precaution to avoid accidental exposure. The patient was maintained with fresh frozen plasma until oral vitamin K1 was obtained. This medication corrected the coagulation disorder. PMID- 11016066 TI - [Current clinical immunology: a genetic and molecular perspective]. AB - During the last few decades, basic scientists and clinicians have gained a deeper insight of the cellular and molecular physiology of the immune system. The widespread application of molecular biology and genetic techniques has advanced our understanding of states of health and disease, bringing forth renewed hopes concerning the advent of a more "specific" therapeutic era of clinical immunology. The precise structural and genetic characterization of molecular complexes such as B and T-cell receptors, the Major Histocompatibility Complex (MHC), cytokines, chemokines, cellular receptors and co-receptors has produced a wealth of information open to both diagnostic and therapeutic purposes. We herein review several recent advances in the molecular and genetic characterization of immune deficiency states, autoimmunity and the induction of antigen specific immune unresponsiveness or tolerance, together with the therapeutic implications of these findings. PMID- 11016067 TI - [Endothelial dysfunction as a primary disorder in vascular diseases]. AB - Endothelium controls vascular smooth muscle tone by secreting relaxing and contracting factors. There is a constant release of endothelium derived relaxing factors, mainly nitric oxide, a potent vasodilator, inhibitor of platelet aggregation, monocyte adhesion and smooth muscle proliferation. In addition, the endothelium may increase the release of NO in response to humoral stimulation by vasoactive substances such as acetylcholine, bradykinin or substance P. Although the endothelium releases a number of products, no single blood test has yet proved useful to determine normal endothelial function or as early abnormalities. The most useful test of endothelial function relies on the measurement of endothelium-dependent dilatation in response to pharmacological or physiologic stimuli. The alteration of this response is known as endothelial dysfunction and has been observed in a variety of circumstances related to cardiovascular risk. This review summarizes the evidence that sustains this association and emphasizes the clinical utility of assessing endothelial function presenting two clinical cases of hypercholesterolemia in which a high-resolution vascular ultrasound in the brachial artery was used. PMID- 11016068 TI - [Academic performance of medical students: a predictable result?]. AB - BACKGROUND: Traditionally, medical schools demand their students a high dedication in time, responsibility and integrity. AIM: To assess the predictive capacity of several specific variables, on the academic performance of medical students. MATERIAL AND METHODS: All students who entered during 1984-1995 period were studied. The academic performance was assessed using two indices: an overall evaluation of successfulness as determined by the approval rate in different courses and grade-point average obtained during the first three years at the Medical School. The variables used to predict academic performance were year of enrollment, high school grades, university admission test scores, biomedical and demographic characteristics. All these were measured at the time when the student was enrolled. RESULTS: Eight hundred and eight students were studied at the end of the third year. The most important predictive variables selected for both performance indices were: high school grades, admission biology test scores, place were high school studies were done, and previous university studies. In addition verbal and mathematics admission academic performance tests scores were selected for grade-point average index. Although, the overall admission score and high school academic performance were significantly associated with the two outcomes, they were not selected in the final models. CONCLUSIONS: The best predictors of an optimal academic performance in these medical students were high school grades, admission biology test scores, residing in Metropolitan Santiago and previous university studies. PMID- 11016069 TI - [UNESCO's bioethical norms to avoid eugenic practices]. AB - The author, member of the UNESCO Bioethics Committee, participated in the preparation of the Universal Declaration about Human Genome and Human Rights, in 1997. The aim of this work is to analyze the initial articles of such Declaration, defining the bioethical principles that defend human dignity, freedom and rights, against the madness of the present biotechnological revolution. The development of genetics for the benefit of mankind will be guaranteed if these principles are honored. Genetic discrimination, reductionism and determinism, are identified by the author as perversions that, if used by biotechnologists, can lead to the rebirth of eugenism and racism, that were condemned by the Code of Nuremberg, in 1947. Investigators must assume their responsibility, respecting the principles of human dignity, the real freedom of research and solidarity among people. This attitude will avoid the use of genetics for purposes other than the welfare of mankind. PMID- 11016070 TI - [Establishing a molecular biology laboratory and standardization of an immunologic methodology in the Regional Clinical Hospital of Valdivia]. PMID- 11016071 TI - [Vancomycin resistant Enterococcus faecium]. PMID- 11016072 TI - Characterization of beta-adrenergic receptors in duck cerebral cortex. AB - beta-adrenoceptor binding sites were characterized in duck cerebral cortex by an in vitro binding technique, using [3H]dihydroalprenolol ([3H]DHA) as a receptor specific radioligand. The specific binding of [3H]DHA to duck cerebral cortical membranes was found to be rapid, stable, saturable, reversible, and of high affinity. Saturation analysis resulted in a linear Scatchard plot suggesting binding to a single class of receptor binding sites with high affinity (Kd = 1.18 nM) and high capacity (Bmax = 162 fmol/mg protein). Competition studies showed the following relative rank order of potency of various compounds to inhibit the [3H]DHA binding: antagonists--ICI 118,551 > S(-)-propranolol >> betaxolol, yohimbine, WB-4101, prazosin, mianserine; agonists--isoprenaline approximately equal to fenoterol > salbutamol >> clonidine, phenylephrine. The obtained data suggest that in duck cerebral cortex beta-adrenergic receptors (like those described in brains of chick and pigeon) are of the beta 2 subtype. This is in contrast to what has been reported for the mammalian brain, where--among beta adrenoceptors--the beta 1 subtype is predominant. PMID- 11016073 TI - Anatomical correlates of the lateral hypothalamic influence on waking-sleep relationship in the rat. AB - Restricted electrolytic lesions of the lateral hypothalamus (LH) evoke sleeplessness in the rat. The present study was aimed to analyze a possible anatomical substrate of the LH hyposomnia within the hypothalamus. In a group of electrolytically lesioned LH rats the intensity of sleep disturbances, assessed on the basis of EEG records from the neocortex and the hippocampus, was confronted with the localization and the extent of destruction of the LH area and with the topography of known fiber systems of the medical forebrain bundle (MFB). In separate experiments the effects of the destruction of LH cell bodies by means of bilateral ibotenic acid (IBO) injections and inhibition of LH neuronal elements by bilateral muscimol (MUSC) administration were also tested. It was found that pronounced hyposomnia follows electrolytic but not IBO lesions of the LH/MFB area. The effective LH damage might have been localized at every level of its antero-posterior axis, from the preoptic area up to the posterior hypothalamus, suggesting involvement of fiber system(s) rather than a localized group of neuronal pericaria. The most effective lesions transsected projections descending from the preoptic/anterior hypothalamic area, olfactory structures, ventral striatum and the central amygdaloid nucleus as well as fibers connecting LH with the brainstem reticular formation, many of them using GABA as a neurotransmitter. Bilateral MUSC injections caused a dose-dependent, bicuculline reversible, increase in waking time, most pronounced at a dose of 50 ng, which ressembled the effect of the electrolytic lesion. These results indicate that LH hyposomnia is not attributable to the damage to the intrahypothalamic neurons and suggest the participation of GABA-ergic transmission in LH in waking-sleep regulation. PMID- 11016074 TI - Spontaneous spike-wave discharges in rat neocortex and their relation to behaviour. AB - A certain proportion of laboratory rats of various strains show spontaneous nonconvulsive ECoG seizures in the form of bursts of spike-and-wave discharges (SWD). Since in the majority of behavioural experiments the EEG is not controlled, the experimenter is usually unaware of this fact. The purpose of the present work was to find out whether the SWD trait is related to the rats behavioural performance in selected test situations. The experiment was performed on two groups of male Wistar rats, outbreds, aged six (group 6M, n = 17) and 24 months (group 24M, n = 14). First, in both groups the following forms of behaviour were assessed: (1) seeking water reward in an 8-arm radial maze, (2) exploration of a new object, (3) inhibition of a locomotor response (passive avoidance), and (4) paw-lick response to a thermal stimulus (54.5 degrees C) applied to the feet before and after intermittent footshock. The rats were then implanted with intrabrain electrodes and the level of SWD activity was assessed. Rats of the 24M group, compared with those of the 6M one, showed a significantly shorter exploratory response to a new object and diminished responsiveness to heat. The groups did not differ, however, in passive avoidance and radial maze performance. The analysis of 3-h ECoG sections revealed SWD bursts in 73% and nearly 93% of rats from groups 6M and 24M, respectively. The groups did not differ in the number of bursts or in the total duration of SWD activity. A correlation analysis of pooled data from both groups revealed that the exploration time of a new object was significantly (negatively) correlated with the number of SWD episodes. The total duration of SWD activity, and the number of perseveration errors in the radial maze, was significantly (positively) correlated with the total duration of SWD activity. The results suggest that SWD rats are behaviourally impaired in some test situations. PMID- 11016075 TI - Bicuculline administration into ventromedial hypothalamus: effects on fear and regional brain monoamines and GABA concentrations in rats. AB - The effects of bicuculline methiodide administration into ventromedial hypothalamus (15 ng per site, bilaterally) on fear behavior and monoamines (NA, DA, 5-HT) and GABA in structures of the brain defensive system (hypothalamus, midbrain gray matter, amygdala, hippocampus and frontal cortex) were studied. Fear behavior was examined in the modified version of light-dark transition test. The time out from the illuminated compartment of chamber, the time spent there and number of returns to the illuminated compartment was measured. Additionally motor activity, i.e., number of crossings and rearings in dark as well as in the illuminated part of compartment, was registered. Blockade of GABAA receptors in the ventromedial hypothalamus resulted in increased fear behavior, i.e. decrease of time out from illuminated compartment and decrease of the time spent there. Motor behavior remained unchanged. HPLC analysis showed reduction of GABA concentration in all investigated brain structures. An increase of NA concentration in all examined structures with exception of the hypothalamus without effect on MHPG/NA was observed as well. Dopamine level remained unchanged, but DOPAC/DA ratio increased in all structures, except frontal cortex. Also HVA/DA ratio increased in the hypothalamus and midbrain. 5-HT concentration increased only in midbrain, 5-HIAA increased in midbrain and in frontal cortex, and 5-HIAA/5-HT ratio increased only in frontal cortex. These results indicate that GABA-ergic and monoaminergic systems remain in functional interactions and that these interactions may play an important role in the neurochemical regulation of fear behavior. The possible mechanism of GABA--monoaminergic interactions is discussed. PMID- 11016076 TI - Pathology and compensation during visual signal processing in schizophrenia- spatial analysis of event-related potentials. AB - Correlations between measures of attention and topographical abnormalities of evoked cortical potentials elicited during the Continuous Attention Test (CAT) were assessed in 50 schizophrenic patients, compared to 50 healthy subjects. For each group and for each CAT condition evoked responses consisted of six successive epochs (segments) of stable spatially configured potentials. Quantitative descriptors of those configurations (Lehmann 1987) were referred to the CAT data. In patients: (1) segments III-V were delayed, (2) in the non-target condition, diminished global field power (GFP) emerged, coexisting either with lower amplitude of posterior potentials in segment I and II or with lower amplitude of a central positive potential in segment V, (3) an altered topographic pattern of responses to the target stimulus occurred. In healthy subjects detection of the target (as compared to the non-target condition) was associated with a shift of the location of the positive potential in segments IV and V from a central towards the prefrontal area. In patients, in segment V a similar shift reached frontal, but not prefrontal areas, and additionally, the central areas remained active. Delayed latency and low GFP in segment V in the non-target condition in patients correlated with poor CAT performance. A more posterior location of the positive centroid in segment V during detection of the target correlated with better CAT results, and the associated GFP increase with less prolonged reaction time. The data revealed a possible compensatory role of central and frontal areas in the face of weakened prefrontal functions in schizophrenia. PMID- 11016077 TI - Effect of arachidonic acid on specific binding of [3H]naloxone to opioid receptors. AB - The effects of arachidonic acid (AA) on binding of [3H]naloxone to the agonist and antagonist configurations of opioid receptors were investigated in rat brain. Equilibrium binding parameters of the agonist and antagonist configurations of the receptors were evaluated from homologue displacement data in the presence or absence of AA. Addition of AA at a concentration of 0.6 mM (1.5 mumole/mg of protein) reduced by 22% and 53% the maximal number of binding sites (Bmax) respectively in the absence or presence of 100 mM NaCl. Binding affinity (KD) was not altered significantly (P < 0.05) either in the presence or absence of 100 mM NaCl and AA. We conclude that AA mediated reduction in [3H]naloxone specific binding was chiefly due to a decrease in the number of binding sites. PMID- 11016078 TI - Oestrogen effects on kainate-induced toxicity in primary cultures of rat cortical neurons. AB - Oestrogens protect neurons against excitatory amino acid-induced toxicity; however data on their interaction with particular subtype of glutamate receptors are sparse. Therefore in the present study we investigated oestrogen effects on kainate neurotoxicity in primary cortical neurons. The data showed that both oestradiol-17 beta and oestrone (100 nM and 200 nM) reduced kainate toxicity by ca. 40%. Since tamoxifen only partly inhibited the above effects, we suggest that both genomic and non-genomic mechanisms are involved in the anti-kainate action of oestrogens. PMID- 11016079 TI - Locomotion induces changes in Trk B receptors in small diameter cells of the spinal cord. AB - INTRODUCTION AND METHODS: Locomotor training leads to improvement of stepping ability in animals after spinal cord transection (1). Recent data point to neurotrophins as possible factors involved in this improvement. Motoneurones synthesising BDNF, NT-4 and NT-3 are a potent source of neurotrophins for the spinal network (2, 3). Physical exercise increases BDNF neurotrophin gene expression in the rat hippocampus (4). If exercise enhances BDNF expression also in the spinal cord, upregulation of its receptor Trk B may occur. To verify this hypothesis we tested whether exercise influences TrkB receptor system in the spinal cord. Six adult, male Wistar rats walked on the treadmill five days a week, 1,000 m daily with the speed of 20 to 25 cm/s. After 4 weeks of training animals were anaesthetised with pentobarbital sodium (80 mg/kg b.w.) and perfused with 0.01 M PBS followed by 2% paraformaldehyde and 0.2% parabenzoquinone in 0.1 M PB. Three non-trained animals were used as controls. Cryostat 40 microns sections were processed free-floating with TrkB polyclonal antibody (1:1,000, Santa Cruz) and ABC Vectastain detection system. Sections were examined under Nikon light microscope and analysed with Image-Pro Plus 4 software. RESULTS AND DISCUSSION: TrkB immunoreactivity (IR) was detected in number of spinal cells at the lumbar level in non-trained animals (Fig. 1A). The strongest IR appeared in the perikarya and processes of small diameter cells rarely scattered in the grey and white matter. The average area of these cells was 50 micron 2 (+/- 10). Exercise increased by over 50% the number of TrkB immunostained small cells (Fig. 1B). An enhancement of perikaryonal immunostaining of these cells was also observed (Fig. 1B, inset). Testing the identity of Trk B IR small diameter cells did not prove their astroglial (GFAP IR) and gabaergic (GAD IR) phenotype in the grey matter. Some of TrkB IR cells in the white matter were astrocytes. Our data point to physical exercise as a potent method to make spinal cells more receptive to neurotrophic stimuli. PMID- 11016080 TI - Severity of insomnia correlates with cognitive impairment. AB - INTRODUCTION AND METHODS: Cognitive deficits in insomnia have been already reported (5), however, a correlation between cognitive impairment and severity of insomnia was not as yet studied. Sixteen not medicated patients with primary insomnia according to DSM-IV (4), 7 men an 9 women, of mean age 40.8 year, were compared to 16 controls, matched according to age, sex and education. Standard polysomnographic data (PSG) were recorded. The next day all the subjects completed Athens Insomnia Scale (AIS) (7), Hyperarousal Scale (HS) (6), Hamilton Depression Rating Scale (HAM-D), Beck Depression Inventory (BDI), Multiple Sleep Latency Test (MSLT) (3), Continuous Attention Test (CAT) with simple reaction time (RT) (8) and Selective Reminding Test (SRT) (2). RESULTS AND DISCUSSION: The psychophysiological differences between patients and controls are shown in Table I. HAM-D and BDI scores were elevated in patients, although none of the patients met clinical criteria of depression. Insomniacs did not differ in the immediate recall, but the number of repetitions necessary to learn all the items of SRT was greater in patients. Insomniacs usually complain of poor performance, however, learning impairment has not been documented in insomnia. Degree of the learning impairment correlated with insomnia score (Fig. 1). Cognitive deficit cannot be due to a daytime sleepiness because sleep latency in all MSLT sessions was not shorter in insomniacs. No correlations between results of SRT and standard PSG parameters were found, in accordance with the thesis that subjective feeling of nonrestorative sleep and other accompanying deficits are only symptoms of an underlying 24-hour disorder (1). PMID- 11016081 TI - MAPK regulation of gene expression in the central nervous system. AB - Long-term potentiation (LTP), a cellular model for long-term memory, is generally acknowledged to consist of both a short-term phase that is characterized by a dependence on autonomous protein kinase activity, and a long-term phase that is characterized by a dependence on changes in gene expression and new protein synthesis. Similarly, long-term memory exhibits a dependence on gene expression and altered protein synthesis. Recent evidence indicates that the mitogen activated protein kinase (MAPK) cascade plays a role in both LTP and long-term memory. The MAPK cascade has heretofore largely been studied in the context of cell division and proliferation and as such, mechanisms for the regulation of gene expression by the MAPK cascade have received considerable attention. Given the possible role of altered gene expression in the late phase of LTP and in long term memory, we evaluated the capacity of the MAPK ERK (extracellular signal regulated kinase) to regulate phosphorylation of the transcription factor cAMP response element binding protein (CREB) in hippocampal area CA1. Our studies indicate a critical role for the MAPK cascade in the regulation of CREB phosphorylation in the hippocampus. PMID- 11016082 TI - Modulation of the composition of AP-1 complex and its impact on transcriptional activity. AB - AP-1 transcription factor known to play a role in cell proliferation and neuronal activation, it is also involved in apoptosis of cells in response to stress, DNA damaging agents or lack of survival signals. AP-1 DNA binding complex is not a single transcription factor but a dimer consisting of members of Fos and Jun families. In this review, we discuss evidence that composition of the AP-1 complex is different under various physiological and pathophysiological conditions. Furthermore, we describe biochemical properties of Fos and Jun proteins that may explain the ability of this transcription factor to activate different sets of genes in response to different stimuli. We propose a hypothesis that AP-1 might contribute to distinct biological processes because an activation of specific signaling pathways results in changes of AP-1 composition and/or phosphorylation status and modulates its transactivating potential towards different promoters. PMID- 11016083 TI - Molecular maps of neural activity and quiescence. AB - The rapid accumulation of inducible transcription factors (ITFs), such as c-Fos and Zif268, in activated neurons combined with histological methods that offer detection at the cellular level are key features that have led to their wide use in visualizing activated neurons. There are two major drawbacks of ITFs that limit their use in the CNS--cell-type expression specificity and stimulus transcription coupling uncertainty. Recent technical advances in the field of molecular activity mapping now permit dual-labeling approaches that help resolve some of these ambiguities and identify neurons that are activated by different sensory stimuli. Furthermore, the recent identification of the robl/LC7-like gene, which shows immediate-early repression after stimulation, may have utility in functional mapping where it can be used to delineate quiescent neurons and serve as a complement to molecular activity markers. PMID- 11016084 TI - Inducing gene expression in barrel cortex--focus on immediate early genes. AB - Using only their vibrissae, rats and mice are able to recognize and differentiate surfaces not distinguishable by primates using their fingertips. It has been shown that sensory stimulation elicits the expression of immediate-early genes (IEG), e.g., c-fos and zif268, in the sensory cortex of rats and mice. Though most of these findings come from visual system, mice and rats rely more on their vibrissal system which also offers many advantages for designing precise and precisely controlled experiments. In this review, new models for the selective and simple stimulation of vibrissae are presented and discussed. The data demonstrating IEG expression in the vibrissal system is also reviewed. PMID- 11016085 TI - Gene expression in learning processes. AB - It has repeatedly been shown that long-term memory formation involves neuronal gene expression. In this article several different roles for neuronal gene function in a context of learning are considered: maintenance of neural functioning, replenishment of cellular elements that are exhausted in response to massive neuronal stimulation accompanying behavioral training, maintenance of the plastically reorganized neuronal connections, and finally integration of information at the level of transcription factor-promoter interaction. It is strongly advocated that only careful scrutiny of learning-related gene expression phenomena may aid in understanding of the complex learning process. PMID- 11016086 TI - Metastable evolutionary dynamics: crossing fitness barriers or escaping via neutral paths? AB - We analytically study the dynamics of evolving populations that exhibit metastability on the level of phenotype or fitness. In constant selective environments, such metastable behavior is caused by two qualitatively different mechanisms. On the one hand, populations may become pinned at a local fitness optimum, being separated from higher-fitness genotypes by a fitness barrier of low-fitness genotypes. On the other hand, the population may only be metastable on the level of phenotype or fitness while, at the same time, diffusing over neutral networks of selectively neutral genotypes. Metastability occurs in this case because the population is separated from higher-fitness genotypes by an entropy barrier: the population must explore large portions of these neutral networks before it discovers a rare connection to fitter phenotypes. We derive analytical expressions for the barrier crossing times in both the fitness barrier and entropy barrier regime. In contrast with 'landscape' evolutionary models, we show that the waiting times to reach higher fitness depend strongly on the width of a fitness barrier and much less on its height. The analysis further shows that crossing entropy barriers is faster by orders of magnitude than fitness barrier crossing. Thus, when populations are trapped in a metastable phenotypic state, they are most likely to escape by crossing an entropy barrier, along a neutral path in genotype space. If no such escape route along a neutral path exists, a population is most likely to cross a fitness barrier where the barrier is narrowest, rather than where the barrier is shallowest. PMID- 11016087 TI - Mixed encounters, limited perception and optimal foraging. AB - This article demonstrates how perceptual constraints of predators and the possibility that predators encounter prey both sequentially (one prey type at a time) and simultaneously (two or more prey types at a time) may influence the predator attack decisions, diet composition and functional response of a behavioural predator-prey system. Individuals of a predator species are assumed to forage optimally on two prey types and to have exact knowledge of prey population numbers (or densities) only in a neighbourhood of their actual spatial location. The system characteristics are inspected by means of a discrete-time, discrete-space, individual-based model of the one-predator-two-prey interaction. Model predictions are compared with ones that have been obtained by assuming only sequential encounters of predators with prey and/or omniscient predators aware of prey population densities in the whole environment. It is shown that the zero-one prey choice rule, optimal for sequential encounters and omniscient predators, shifts to abruptly changing partial preferences for both prey types in the case of omniscient predators faced with both types of prey encounters. The latter, in turn, become gradually changing partial preferences when predator omniscience is considered only local. PMID- 11016088 TI - Modeling immune responses with handling time. AB - Simple predator-prey type models have brought much insight into the dynamics of both nonspecific and antigen-specific immune responses. However, until now most attention has been focused on examining how the dynamics of interactions between the parasite and the immune system depends on the nature of the function describing the rate of activation or proliferation of immune cells in response to the parasite. In this paper we focus on the term describing the killing of the parasite by cell-mediated immune responses. This term has previously been assumed to be a simple mass-action term dependent solely on the product of the densities of the parasite and the immune cells and does not take into account a handling time (which we define as the time of interaction between an immune cell and its target, during which the immune cell cannot interact with and/or destroy additional targets). We show how the handling time (i) can be incorporated into simple models of nonspecific and specific immunity and (ii) how it affects the dynamics of both nonspecific and antigen-specific immune responses, and in particular the ability of the immune response to control the infection. PMID- 11016089 TI - Diffusion of clusters of transmembrane proteins as a model of focal adhesion remodeling. AB - Focal adhesions play a major role in maintaining the cell shape and motility, and in regulating numerous cellular processes. Observations suggest that the functioning of focal adhesions is possible due to their dynamic nature, yet the mechanisms that govern their motion are not well understood. This study addresses the process of focal adhesion remodeling using two distinct theoretical approaches. Namely, adhesion sites are modeled as clusters of integrins that are either bound to cytoskeletal elements or dissociated and temporarily free of any attachments. In the first approach effects of cluster size and permeability on the diffusion of mobile adhesion structures are studied using Brinkman's effective medium approach. Diffusion coefficients calculated by this hydrodynamic model significantly decrease with the increase in contact area (the effective size of the focal adhesion). In the second approach focal adhesions are modeled as clusters of transmembrane proteins tightly connected to the cytoskeleton, but still capable of motion. The remodeling of these clusters is coupled to the deformation of the cytoskeleton by means of equating energies at the end states of a reversible elastodynamic interaction. Due to large uncertainty of the plasma membrane and the cytoskeleton properties, predicted diffusion coefficients vary within several orders of magnitude. However, a reasonable set of parameters for each model yields diffusion coefficients that compare favorably with those measured by single-particle tracking (SPT), fluorescence recovery after photobleaching (FRAP), and fluorescence digital imaging (FDI). The estimated Young's modulus of the stress fibers is also in good agreement with measurements. To assess the relevance of the models to focal adhesion remodeling and to improve their predictions, further data on the morphology of focal adhesions and on properties of the plasma membrane and the cytoskeleton are required. PMID- 11016090 TI - A measure of sexual dimorphism in populations which are univariate normal mixtures. AB - Measures of sexual dimorphism have been used extensively to predict the social organization and ecology of animal and human populations. There is, however, no universally accepted measure of phenotypic differences between the sexes. Most indices of sexual dimorphism fail to incorporate all of the information contained in a random data set. In an attempt to have a better alternative, an index is proposed to measure sexual dimorphism in populations that are distributed according to a probabilistic mixture model with two normal components. The index calculates the overlap between two functions that represent the contribution of each sex in the mixture. In order to assess such an index, sample means, variances and sizes of each sex are needed. As a consequence, the sample information used is greater than that used by other indices that take intrasexual variability into account. By evaluating some examples, our proposed index appears to be a more realistic measure of sexual dimorphism than other measures currently used. PMID- 11016091 TI - Local populations of different sizes, mechanistic rescue effect and patch preference in the Levins metapopulation model. AB - In this paper three extensions of the Levins metapopulation model are discussed: (1) It is shown that the Levins model is still valid if patches contain local populations of different sizes with different colonization and extinction rates. (2) A more mechanistic formulation of the rescue effect is presented. (3) The addition of preference of dispersers for occupied or empty patches and its consequences for conservation strategies are studied. PMID- 11016092 TI - Spatial structure and fluctuations in the contact process and related models. AB - The contact process is used as a simple spatial model in many disciplines, yet because of the buildup of spatial correlations, its dynamics remain difficult to capture analytically. We introduce an empirically based, approximate method of characterizing the spatial correlations with only a single adjustable parameter. This approximation allows us to recast the contact process in terms of a stochastic birth-death process, converting a spatiotemporal problem into a simpler temporal one. We obtain considerably more accurate predictions of equilibrium population than those given by pair approximations, as well as good predictions of population variance and first passage time distributions to a given (low) threshold. A similar approach is applicable to any model with a combination of global and nearest-neighbor interactions. PMID- 11016093 TI - Seasonal temperature alone can synchronize life cycles. AB - In this paper we discuss the effects of yearly temperature variation on the development and seasonal occurrence of poikiliothermic organisms with multiple life stages. The study of voltinism in the mountain pine beetle (Dendroctonus ponderosae Hopkins), an important forest insect living in extreme temperature environments and exhibiting no diapause, provides a motivational example. Using a minimal model for the rates of aging it is shown that seasonal temperature variation and minimal stage-specific differences in rates of aging are sufficient to create stable uni- and multi-voltine oviposition cycles. In fact, these cycles are attracting and therefore provide an exogenous mechanism for synchronizing whole populations of organisms. Structural stability arguments are used to extend the results to more general life systems. PMID- 11016094 TI - Luminaries in medicine: Richard Mead, James Gibbs, and solar and lunar effects on the human body in early modern England. PMID- 11016095 TI - The disease of the self: representing consumption, 1700-1830. PMID- 11016096 TI - The holistic gaze in German medicine, 1890-1930. PMID- 11016097 TI - "The eyes have it": trachoma, the perception of disease, the United States Public Health Service, and the American Jewish immigration experience, 1897-1924. PMID- 11016099 TI - Netnotes: medical history on the Internet. Ping Me. PMID- 11016098 TI - In memoriam: Mirko Drazen Grmek, 9 January 1924-6 March 2000. PMID- 11016100 TI - Evaluating return-on-investment for a hospital clinical information system. PMID- 11016101 TI - Measurement of the effects of an integrated, point-of-care computer system on quality of nursing documentation and patient satisfaction. AB - This quasi-experimental, modified time series study measured the effects of the nursing module of a point of care clinical information system on nursing documentation and patient satisfaction. Measurements were taken before implementation of the module and at 6-, 12-, and 18-month intervals postimplementation. Quality of nursing documentation was measured by compliance to items applicable to nursing documentation selected from the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) Closed Medical Review Tool. Patient satisfaction was measured by using the Risser Patient Satisfaction Scale. The study data showed a statistically significant increase in the quality of nursing documentation after implementation of the computerized nursing documentation system, as well as a decrease in variability in charting, as evidenced by a decrease in standard deviations. A significant increase in charting compliance was still occurring between the 12- and the 18-month time points after initiation of automated documentation. The point of care computer system did not seem to affect patient satisfaction with the nurse-patient relationship. PMID- 11016102 TI - Publishing a nursing textbook: collaborating through "seamless technology". AB - Electronic communication can enhance long-distance networking and scholarly activity and make long-distance collaboration through technology a reality. Technology presents a new world of opportunity for improving writing skills, nurturing collegiality, and maximizing efficiency in the successful completion of collaborative projects. The concept of "seamless technology" introduced by the authors provides a guide to assist partners involved in collaborative projects to manage technology for a successful, rewarding experience. Basic questions regarding technology that need to be answered on the front end of a long-distance project are suggested in this article. In addition, the steps taken by the authors to co-author and co-edit a nursing textbook and accompanying instructor's manual by using electronic communication and other sources of technology are outlined. The authors address ways that long-distance collaboration required them to maximize their communication and planning skills, thereby increasing productivity and reducing costs. The challenges and rewards of working together exclusively through technology are discussed, and finally, implications for professional nursing are addressed. PMID- 11016103 TI - Introduction of nursing informatics in the nursing baccalaureate program at the American University of Beirut. AB - The rapid expansion of computer use for various nursing activities has made computer technology an important part of the curriculum in many schools of nursing across the world. Computer technology has become an alternative tool for teaching practice skills to students in a variety of educational settings and disciplines. Information technology (IT) is not yet well recognized in the Lebanese nursing curricula. This article traces the events of planning, developing, integrating, and evaluating nursing informatics on the Bachelor of Science in Nursing (BSN) Program of the School of Nursing (SON), at The American University of Beirut (AUB), Lebanon. PMID- 11016104 TI - Functional health pattern assessment on-line. Lessons learned. AB - Assessment of patient care needs is the foundation for planning patient care and documenting the nursing process to enhance the nurse's ability to coordinate care, plan discharges effectively, and provide appropriate patient/family education. An on-line assessment module in the hospital clinical information system was implemented. This module uses Gordon's Functional Health Pattern Assessment as the foundation for the professional nursing documentation system in this academic, health science center. The article provides a description of the design of the pathways for the on-line assessment, analysis of the design, benefits of using the on-line assessment, and lessons learned from the design and implementation of the pathways. PMID- 11016105 TI - Cognitive processing of trauma cues in adults reporting repressed, recovered, or continuous memories of childhood sexual abuse. AB - Psychologically traumatized people exhibit delayed color naming of trauma words in the emotional Stroop task. Four groups of participants were asked to color name positive words, neutral words, and trauma words; these groups included 15 women who believed that they harbored repressed memories of childhood sexual abuse (CSA), 13 women who reported recovered memories of CSA, 15 women who had never forgotten their CSA, and 12 women who had never been abused. Repressed memory participants exhibited patterns of interference indistinguishable from those of the nonabused control group participants. Irrespective of group membership, the severity of self-reported posttraumatic stress disorder symptoms was the only significant predictor of trauma-related interference, r(48) = .30, p < .05. PMID- 11016106 TI - A study of genetic and environmental effects on the co-occurrence of problem behaviors in three-year-old twins. AB - The authors examined the genetic and environmental causes of the co-occurrence of problem behaviors in children. The analyses involved mother and father ratings of Oppositional, Withdrawn/Depressed, Aggressive, Anxious, Overactive, and Sleep Problems in 446 monozygotic and 912 dizygotic pairs of 3-year-old twins. Genetic factors contributed on average .150 (37.3%), shared environment .206 (51.2%), and nonshared environment .046 (11.4%) to the phenotypic correlations between the syndromes. Genetic and environmental factors caused different groupings. Internalizing and Externalizing groupings were indicative of nonshared environmental factors; clusters of problem behaviors with either the Aggressive or Anxious symptoms were most suggestive of genetic factors, and high scores on all syndromes indicated shared environmental influences. PMID- 11016107 TI - The psychopath as observer: emotion and attention in picture processing. AB - This study extended prior work showing abnormal affect-startle modulation in psychopaths. Male prisoners viewed specific categories of pleasant (erotic or thrilling) and unpleasant (victim or direct threat) slide pictures, along with neutral pictures. Acoustic startle probes were presented early (300 and 800 ms) and late (1,800, 3,000, and 4,500 ms) in the viewing interval. At later times, nonpsychopaths showed moderate and strong reflex potentiation for victim and threat scenes, respectively. For psychopaths, startle was inhibited during victim scenes and only weakly potentiated during threat. Psychopaths also showed more reliable blink inhibition across pleasant contents than nonpsychopaths and greater heart rate orienting to affective pictures overall. These results indicate a heightened aversion threshold in psychopaths. In addition, deficient reflex modulation at early times suggested a weakness in initial stimulus evaluation among psychopaths. PMID- 11016108 TI - The relationship between cognitive inhibition and psychotic symptoms. AB - Cognitive models of schizophrenia have highlighted deficits of inhibitory attentional processes as central to the disorder. This has been investigated using "negative priming" (S. P. Tipper, 1985), with schizophrenia patients showing a reduction of negative priming in a number of studies. This study attempted to replicate these findings, but studied psychotic symptoms rather than the broad diagnostic category of schizophrenia. Psychotic individuals exhibiting positive symptoms were compared with asymptomatic psychiatric patients and with a normal control group. As predicted, the symptomatic group failed to show the usual negative priming effect, which was present in the asymptomatic and normal groups. A modest but significant correlation was found between negative priming and delusions. Neither diagnosis, nor affective or negative symptoms, nor chronicity, nor medication, was related to negative priming. These data replicate previous findings that positive symptoms are related to a reduction in cognitive inhibition, although considerable variability was observed among the psychotic patients. PMID- 11016109 TI - Cognitive abilities in adolescent-limited and life-course-persistent criminal offenders. AB - T. E. Moffitt's (1993a) hypothesis that adolescent-limited criminal offenders will have higher scores on tests of cognitive ability than life-course-persistent offenders was tested with 12 tests of cognitive ability given to a large and diverse sample of delinquent juveniles whose arrest records were collected over 20 years. This is the first investigation to empirically evaluate this proposal with longitudinal data obtained from a sample for a long enough time to distinguish life course patterns of crime. This study provided only partial support for Moffitt's hypothesis because the results varied by ethnicity. We found relatively consistent support for the hypothesis for Caucasians and Hispanics but no support for the hypothesis for African Americans. These findings are interpreted in terms of differences in developmental contexts for individual ethnic groups. PMID- 11016110 TI - The Temple-Wisconsin Cognitive Vulnerability to Depression Project: lifetime history of axis I psychopathology in individuals at high and low cognitive risk for depression. AB - The authors tested the cognitive vulnerability hypotheses of depression with a retrospective behavioral high-risk design. Individuals without current Axis I diagnoses who exhibited either negative or positive cognitive styles were compared on lifetime prevalence of depressive and other disorders and the clinical parameters of depressive episodes. Consistent with predictions, cognitively high-risk participants had higher lifetime prevalence than low-risk participants of major and hopelessness depression and marginally higher prevalence of minor depression. These group differences were specific to depressive disorders. The high-risk group also had more severe depressions than the low-risk group, but not longer duration or earlier onset depressions. The risk group differences in prevalence of depressive disorders were not mediated by current depressive symptoms. PMID- 11016111 TI - Comparison of DSM-III-R chronic major depression and major depression superimposed on dysthymia (double depression): validity of the distinction. AB - The nosology of chronic depression has become increasingly complex since the publication of the revised third edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R; American Psychiatric Association, 1987), but there are few data available to evaluate the validity of the distinctions between the subtypes of chronic depression. The validity of the distinction between DSM III-R chronic major depression (CMD) and major depression superimposed on dysthymia (double depression, DD) was examined. Participants were 635 patients with chronic depression in a 12-week trial of antidepressant medications. Patients with CMD, DD, and a 3rd group with a chronic major depressive episode superimposed on dysthymia (DD/CMD) were compared on demographic and clinical characteristics, family history, and response to treatment. Few differences were evident, although the depression of patients with DD/CMD tended to be more severe. PMID- 11016112 TI - Common and specific correlates of preadolescent internalizing and externalizing psychopathology. AB - The specificity of various child characteristics and environmental correlates of childhood internalizing and externalizing problems was examined using both cross sectional and longitudinal analyses (from ages 2-3 and 4-5 years) in a general population sample of 10-11-year-olds. Specificity was defined according to a between-subjects and a within-subjects method, using parent and teacher reports of psychopathology. Temperamental withdrawal, parental internalizing psychopathology, and early single parenthood (for girls) were identified as correlates that are specific for internalizing problems, whereas temperamental high general activity level was identified as externalizing-specific. Further, parenting stress, poor school results (only for boys), and stressful life events (only for girls) were found to be common correlates of psychopathology. Research implications regarding the findings and the use of a within-subjects method are discussed. PMID- 11016113 TI - Body-image and eating disturbances predict onset of depression among female adolescents: a longitudinal study. AB - This study examined data from a 4-year school-based longitudinal study (n = 1,124), to test whether the increase in major depression that occurs among girls during adolescence may be partially explained by the body-image and eating disturbances that emerge after puberty. Elevated body dissatisfaction, dietary restraint, and bulimic symptoms at study entry predicted onset of subsequent depression among initially nondepressed youth in bivariate analyses controlling for initial depressive symptoms. Although the unique effect for body dissatisfaction was not significant in the multivariate model, this set of risk factors was able to fairly accurately foretell which girls would go on to develop major depression. Results were consistent with the assertion that the body-image- and eating-related risk factors that emerge after puberty might contribute to the elevated rates of depression for adolescent girls. PMID- 11016114 TI - Recognition of posed and genuine facial expressions of emotion in paranoid and nonparanoid schizophrenia. AB - Most previous research reporting emotion-recognition deficits in schizophrenia has used posed facial expressions of emotion and chronic-schizophrenia patients. In contrast, the present research examined the ability of patients with acute paranoid and nonparanoid (disorganized) schizophrenia to recognize genuine as well as posed facial expressions of emotion. Evidence of an emotion-recognition deficit in schizophrenia was replicated, but only when posed facial expressions were used. For genuine expressions of emotion, the paranoid-schizophrenia group was more accurate than controls, nonparanoid-schizophrenia patients, and depressed patients. Future research clearly needs to consider the posed versus genuine nature of the emotional stimuli used and the type of schizophrenia patients examined. PMID- 11016115 TI - Borderline personality disorder symptoms as predictors of 4-year romantic relationship dysfunction in young women: addressing issues of specificity. AB - The relationships between romantic relationship dysfunction and symptoms of borderline personality disorder (BPD), other personality disorders, and depression were examined prospectively in a community sample of 142 late adolescent women. Although BPD symptoms predicted 4-year romantic dysfunction (romantic chronic stress, conflicts, partner satisfaction, abuse, and unwanted pregnancy), the associations were not unique to BPD. Instead, relationship dysfunction was better predicted by a cumulative index of non-BPD Axis II pathology. Depression did not predict outcomes uniquely when Axis II symptoms were included, except in the case of unplanned pregnancy. The results suggest that although BPD is associated with relationship dysfunction, the effect is a more general phenomenon applying rather broadly to Axis II pathology. The results also highlight the importance of subclinical psychopathology in the construction of early intimate relationships. PMID- 11016116 TI - Verbal processing deficits in schizophrenia. AB - The authors reported that a subgroup of schizophrenic patients performed well on a tone serial position task but was impaired on an auditory word serial position task (Wexler, Stevens, Bowers, Cerniak, & Goldman-Rakic, 1998). This study assessed 30 schizophrenic and 32 controls (matched for comparable tone discrimination) on 4 versions of the verbal serial position tasks and 2 tone serial position tasks. Patients performed poorly on all verbal tasks but performed comparably to controls when tones served as stimuli. Proactive interference and visual presentation further compounded the verbal deficits. Deficits persisted with pronounceable nonword stimuli. These findings provide evidence of specific deficits in language-related processing, although the authors could not rule out the possibility that the differential effects that were observed between the tone and word tasks, and particularly among the verbal tasks, may result from differing discriminating power of the different tests. PMID- 11016117 TI - Informing the continuity controversy: a taxometric analysis of depression. AB - Researchers and practitioners have long debated the structural nature of mental disorders. Until recently, arguments favoring categorical or dimensional conceptualizations have been based primarily on theoretical speculation and indirect empirical evidence. Within the depression literature, methodological limitations of past studies have hindered their capacity to inform this important controversy. Two studies were conducted using MAXCOV and MAMBAC, taxometric procedures expressly designed to assess the underlying structure of a psychological construct. Analyses were performed in large clinical samples with high base rates of major depression and a broad range of depressive symptom severity. Results of both studies, drawing on 3 widely used measures of depression, corroborated the dimensionality of depression. Implications for the conceptualization, investigation, and assessment of depression are discussed. PMID- 11016119 TI - The role of rumination in depressive disorders and mixed anxiety/depressive symptoms. AB - Several studies have shown that people who engage in ruminative responses to depressive symptoms have higher levels of depressive symptoms over time, after accounting for baseline levels of depressive symptoms. The analyses reported here showed that rumination also predicted depressive disorders, including new onsets of depressive episodes. Rumination predicted chronicity of depressive disorders before accounting for the effects of baseline depressive symptoms but not after accounting for the effects of baseline depressive symptoms. Rumination also predicted anxiety symptoms and may be particularly characteristic of people with mixed anxiety/depressive symptoms. PMID- 11016118 TI - Age and gender differences in oppositional behavior and conduct problems: a cross sectional household study of middle childhood and adolescence. AB - Behavior problems among youths cannot be understood without explaining their age and gender differences, but age and gender differences cannot be explained until they have been accurately described. In a household survey of 1,285 youths aged 9 to 17 years, there were no gender differences in oppositional behavior, but aggression, property offenses, and status offenses were more common among boys. Levels of oppositional behavior were greater at younger ages, aggression peaked near the middle of this age range, and property and status offenses were more prevalent at older ages. These findings are generally consistent with developmental models of conduct problems but are inconsistent with a recent model of gender differences and raise questions about the external validity of current taxonomies. PMID- 11016120 TI - Emotion recognition in schizophrenia: further investigation of generalized versus specific deficit models. AB - In this study, the authors examined the nature of emotion perception in schizophrenia. Two samples of people with schizophrenia, one receiving acute care for a recent exacerbation of symptoms and the other receiving extended care, were compared with a nonclinical control group on emotion perception and general perception measures. The nonclinical control group obtained the highest scores on all of the study measures, and the acutely ill group obtained the lowest scores. Furthermore, the acutely ill sample had a specific deficit in emotion perception that remained present after controlling for performance on the general perception tasks. Conversely, the deficits in emotion discrimination in the extended-care sample reflected generalized poor performance. Differences in performance on the emotion identification task between the 2 clinical groups were reduced when controlling for active symptoms. PMID- 11016121 TI - Are trauma victims susceptible to "false memories"? AB - Laboratory studies using word-list paradigms have provided evidence that nontraumatized individuals falsely recall or recognize events that never occurred. In the present study, H. L. Roediger and K. B. McDermott's false-memory paradigm (1995) was utilized to examine possible source monitoring deficits in individuals with PTSD. Traumatized individuals with PTSD were compared with traumatized individuals without PTSD and with nontraumatized control participants. Participants heard lists of related words (e.g., bed, night) that were associates of a critical nonpresented word (e.g., sleep) and were given immediate free recall and later recognition tests. Traumatized participants with and without PTSD generated more false recalls of critical nonpresented words than did nontraumatized participants. False recall was related to trait anxiety and PTSD severity. The results are consistent with a general source-monitoring deficit in trauma-exposed individuals. PMID- 11016122 TI - Predictors of first onset and recurrence of major depression in young women during the 5 years following high school graduation. AB - Although most depressive episodes in adulthood are recurrences of the disorder, lifetime history of major depression (MD) is often neglected in predictive models. On the basis of research and theory suggesting differential prediction of MD across the course of the disorder, the authors explored whether factors that predict a first MD onset would not predict MD recurrence. Predictors of MD were examined longitudinally in a sample of 128 young women followed for 5 years. Controlling for lifetime MD history, 5-year MD was predicted by the presence before study entry of 3 variables: having witnessed family violence before age 16, having a parent with a psychiatric disorder, and having a nonmood Axis I disorder. During the follow-up period, chronic and episodic stress predicted MD. Prior lifetime MD interacted with both chronic stress and parental psychopathology to predict MD, such that first onsets, but not recurrences, were predicted by these risk variables. PMID- 11016123 TI - Learning and forgetting in schizophrenia. AB - Recent studies of patients with schizophrenia have consistently demonstrated marked deficits on measures of initial learning. However, contradictory results have been reported concerning retention and forgetting. The present study examined the level of initial and delayed recall of stories and visual figures in a group of 76 patients with schizophrenia and 51 normal controls. Schizophrenia patients demonstrated marked impairments in initial and delayed recall as well as significantly worse percentage retention scores. However, schizophrenia patients and healthy controls individually matched on level of initial recall had nearly identical delayed recall performance. The results suggest a primary deficit in the initial acquisition of information rather than an accelerated rate of forgetting in schizophrenia. PMID- 11016124 TI - Personality and family functioning in families of depressed patients. AB - In this cross-sectional study, the authors attempted to identify correlates of family functioning in 86 couples with a depressed member during the acute phase of the patient's depression. Demographic variables, psychiatric status, and personality traits of both the patient and spouse were investigated as potential predictors of family functioning. Regression analyses indicated that lower levels of personality pathology in the patient, higher levels of patient conscientiousness, and less psychological distress in the spouse were associated with healthier family functioning. Future research implications and clinical importance of these findings are discussed. PMID- 11016125 TI - Dichotic listening in college students who report auditory hallucinations. AB - College students were placed in 2 groups, hallucinators and nonhallucinators, on the basis of their responses to a verbal hallucinations questionnaire. Both groups were given a consonant-vowel version of a Dichotic Listening Test under 3 conditions: nonforced, forced-right, and forced-left. When hallucinators were instructed to attend to the left ear stimuli (forced-left condition), they had fewer correct responses to right ear syllables than did nonhallucinators. This resulted in a left ear advantage for hallucinators. When nonhallucinators were instructed to attend to the left ear, they maintained a right ear advantage. Results suggest that auditory hallucinations in college students are associated with differences in hemispheric functioning. PMID- 11016126 TI - Predicting negative spousal attitudes toward depressed persons: a test of Coyne's interpersonal model. AB - The utility of Coyne's (1976a) interactional model in predicting negative spousal attitudes toward depressed patients was examined. Eighty-nine couples with at least 1 member in treatment for depression were selected on the basis of semistructured diagnostic interviews. Overall, spouses living with a depressed patient reported significantly more distress than population norms. Consistent with prediction, patients' reassurance seeking and spouses' mood contributed to negative spousal attitudes. Additional analyses demonstrated that these effects persisted even after controlling for spouse marital adjustment, suggesting that negative spousal attitudes were more than a simple reflection of marital maladjustment. PMID- 11016127 TI - Fantasy proneness, dissociation, and DSM-IV axis II symptomatology. AB - Scores on the Inventory of Childhood Memories and Imaginings (S. C. Wilson & T. X. Barber, 1981) identified fantasizers and controls who were administered the Structured Interview for DSM-IV Personality (SIDP-IV; B. Pfohl, N. Blum, & M. Zimmerman, 1994) and the Dissociative Experiences Scale (DES; E. M. Bernstein & F. W. Putnam, 1986). Fantasizers had increased rates of Cluster A and B personality disorders but were equivalent to controls in Cluster C diagnoses. For both Clusters A and B, 55% of the fantasizers received clinical diagnoses. Fantasizers had significantly higher DES, DES-T (N. G. Waller, F. W. Putnam, & E. B. Carlson, 1996), and Normal Dissociative Index scores. Although nearly one half of the fantasizers' DES-T patterns were classified within the pathological dissociative taxon (N. G. Waller & C. A. Ross, 1997), none of the controls and only 10.03% of the original screening sample received this classification. Both Axis II pathology and pathological dissociation were associated with fantasy proneness. PMID- 11016128 TI - Social norms research in college health. PMID- 11016129 TI - College students define binge drinking and estimate its prevalence: results of a national survey. AB - Data from the 1999 College Alcohol Study were used to examine how students define the term binge drinking, to determine how much binge drinking the students think exists on their campuses, and to analyze how students' estimates compare with aggregated self-reports of student drinking. The findings indicate that the median of the students' definitions of binge drinking is 6 drinks in a row for men and 5 for women, 1 drink higher than the definition used by researchers. Students' definitions of binge drinking vary with their own drinking levels, suggesting that dissenting views of the research definition may represent voices of the heaviest drinkers. At the median, students estimated that 35% of all students were binge drinkers. Half (47%) of the students underestimated the binge drinking rate at their school, 29% overestimated it, and 13% were accurate. Although programs designed to reduce the frequency or prevalence of binge drinking by emphasizing healthier norms would be most useful in addressing binge drinkers who overestimate drinking norms, this group includes only 13% of college students. PMID- 11016130 TI - The efficacy of the social norms approach to substance abuse prevention applied to fraternity men. AB - Students tend to overestimate the amount of alcohol consumed among their peers and often drink to that imaginary level. The social norms strategy, designed to correct norm misperceptions, has been correlated with a decrease in reported consumption in the general college population. However, it has had little or no impact among Greek students, the group that consumes the most alcohol. The authors investigated and subsequently found three possible flaws in the application of the social norms strategy that may account for the failure to decrease binge drinking among fraternity men: there is no predominant, healthy drinking norm in this population; students are influenced more by people within their network(s) than by others; and binge drinking is the norm in this group and may serve to perpetuate the problem. The findings, though preliminary, provide the first step in developing interventions beyond the social norms approach to address binge drinking among fraternity men. PMID- 11016131 TI - A test of the perceived norms model to explain drinking patterns among university student athletes. AB - The author tested the ability of perceived drinking norms to discriminate among drinking patterns in a sample of National Collegiate Athletic Association (NCAA) Division I student athletes. He used an anonymous questionnaire to assess 297 athletes, representing 18 teams, at a public university in the Midwest. Alcohol use patterns showed considerable variation, with many athletes (37.1%) abstaining during their season of competition. A discriminant function analysis revealed that higher levels of alcohol involvement are disproportionately found among athletes who began drinking regularly at an early age. Perceived drinking norms were less important in the discrimination of student athlete drinker groups. Women and those with higher grade point averages were somewhat more likely to refrain from in-season drinking than other survey respondents. PMID- 11016132 TI - Results of a social norm intervention to prevent binge drinking among first-year residential college students. AB - The effects of a primary prevention social norm intervention on binge drinking among 1st-year residential college students were examined. Six hundred thirty four students attending a medium-sized public university in the South were randomly assigned to receive a two-phase social norm intervention or the standard campus psychoeducational prevention program. At posttest, no differences were found between intervention and control group students on any of the alcohol use and alcohol-use risk factor measures. Significant subgroup differences were found by stage of initiating binge drinking behaviors, for frequency of alcohol use, F(3, 507) = 3.69, p = .01; quantity of alcohol use, F(3, 507) = 2.51, p = .05; and social norms, F(3, 505) = 2.53, p = .05. These findings suggest the need for tailoring social norm binge drinking interventions to students' stage of initiating heavy drinking and carefully monitoring for potential negative, as well as positive, effects of norm-based prevention messages. PMID- 11016133 TI - Developing a college men's growth group. AB - The higher rates of disease and death associated with male gender have been widely documented, yet little is being done in the college health setting to address men's health issues. Many of men's health risks are related to social isolation and prohibitions on showing weakness. The authors describe the Virginia Tech University experience with 3 Man Alive groups that were developed to (a) create a supportive social structure for male college students who may otherwise be socially isolated, (b) model healthy expression of emotions, such as anger, fear, sadness, and joy; (c) create a safe space for men to discuss sensitive issues, such as relationships with significant others, sexuality, and substance abuse. PMID- 11016134 TI - The national programme and the IMA. PMID- 11016135 TI - Poliomyelitis on the verge of blotting out. PMID- 11016136 TI - Factors responsible for delayed immunisation among children under 5 years of age. AB - Although immunisation coverage has increased substantially in recent years, still a sizable proportion of children are not being immunised at appropriate time. The present hospital based, pair matched, case control study was carried out at immunoprophylactic centre of Government Medical College and Hospital, Nagpur, to identify risk factors associated with delayed immunisation among children. This study included 139 children with a delay of more than 90 days for scheduled primary immunisation and equal number of matched control (for dose) receiving immunisation at appropriate time. The study identified significant association of family size [Odd's ratio (OR) = 7.3, 95% confidence interval (CI) = 1.5-35.6], number of children < 5 years (OR = 3.17, 95% CI = 1.1-9.9), sex (OR = 3, 95% CI = 1.2-7.4), paternal education (OR = 3.7, 95% CI = 1.6-8.5), maternal education (OR = 4, 95% CI = 1.5-10.9), socio-economic status (OR = 3.7, 95% CI = 1.1-13.2) and distance from health centre (OR = 4.7, 95% CI = 1.2-17.6) with delayed immunisation. Negligence (56%) and unawareness (22.7%) of parents were main reasons for delayed immunisation. Hence more stress on identified risk factors in the study will indirectly help in reducing the frequency of delayed immunisation. PMID- 11016137 TI - Surveillance for polio eradication: current status and lessons learnt--India, 1999. AB - With the launch of the Universal Immunisation Programme in India in 1985, childhood immunisation was provided to children in all districts of the country in a phased manner by 1990. Surveillance for vaccine preventable diseases (VPD) including polio was started at the same time with monthly reporting from the districts to the Ministry of Health and Family Welfare (MOHFW), Government of India (GOI). In 1995, the Pulse Polio Immunisation (PPI) campaign was launched with the objective of polio eradication. Prior to 1997, surveillance for polio was directed at finding clinical polio cases by passive reporting from health facilities. There was no active surveillance for all cases of acute flaccid paralysis (AFP). In 1996, a scheme for the surveillance of AFP was drawn up. With the support of the Danish and US governments and Rotary International, 59 surveillance medical officers (SMOs) were hired, trained, and posted throughout the country in October 1997 to establish active surveillance of AFP. The number of SMOs was increased to 108 in August 1999. The SMOs along with their government counterparts established 10,069 reporting units nationwide by the end of November 1999 reporting weekly the occurrence of AFP cases to the district, state, and national levels; timely case investigation and collection of stool specimens from AFP cases; linkages to support the polio laboratory network; and extensive training of government counterparts. Data reported to the national level is analysed and put on an internet website which is updated every two weeks. Annualised rates of reported non-polio AFP have increased from 0.22 per 100,000 children aged < 15 years in 1997 to 1.57 in 1999. The number of polio cases associated with isolation of wild poliovirus decreased from 1404 in the third trimester of 1998 to 664 in the third trimester of 1999, yet widespread transmission of wild polioviruses persists throughout the country. PMID- 11016138 TI - Trends and determinants of immunisation coverage in India. AB - The 20th century has witnessed many important events in the control of infectious diseases that mostly affect children. In addition to the eradication of smallpox, the interruption of poliomyelitis transmission in many countries with a distinct possibility of its eradication by the turn of this century are some of the major achievements. Also, the rates of other vaccine preventable diseases such as measles, pertussis and diphtheria have gone down significantly. The discovery and use of vaccines have made it possible to save approximately 8 million deaths, annually. This is in addition to the reduction in millions of children's suffering and disability. It is now important to build on these gains through adequate utilisation of other vaccines e.g., hepatitis B, typhoid and Haemophilus influenzae type b that are currently available, but in limited use. But, a high level of coverage for any vaccination programme is a pre-requisite to witness the effective reduction of the specific disease against which child population is vaccinated. This paper reviews the coverage levels by surveys in the last 3 years. It has been observed that vaccination coverage levels are falling. Keeping the promises of immunising every child to fulfill his/her right is the need of the hour. To achieve this the major action points are: (a) The need for organising fixed immunisation sessions at the community, where low proportion of sessions are held; and (b) The need to improve demand generation activities where the coverage is poor despite better service availability at the community level. Therefore, the challenge for the next century is to make sure that the enormous impact of vaccines on the health and well-being of the population is maintained as well as expanded. Vaccines that effectively prevent rotavirus diarrhoea, pneumococcal pneumonia, menigococcal meningitis, if made available, could prevent deaths up to two million a year. Research efforts are currently under progress to develop new vaccines against malaria, tuberculosis, shigella-induced dysentery, and Esch coli-induced diarrhoea. PMID- 11016139 TI - The global poliomyelitis eradication initiative: progress and challenges. AB - In 1988, The World Health Assembly committed WHO and its member states to the goal of poliomyelitis eradication by the year 2000. Global progress in implementation of strategies include routine and supplementary immunisation, AFP surveillance strategy and mopping up. Progress made in global polio eradication within 10 years has been dramatic. Challenges consist of tailoring and fine tuning strategies and sustaining adequate levels of findings. Although the intensified effort will increase needed resources in the short term, it will save costs in the long term. PMID- 11016140 TI - Lessons learnt from pulse polio immunisation programme. AIIMS-India CLEN PPI Program Evaluation 1997-98 Team. PMID- 11016142 TI - Postpolio syndrome. PMID- 11016141 TI - Vaccines: quality issues. PMID- 11016144 TI - Myths about vaccination. PMID- 11016143 TI - Immunisation and the cold chain. PMID- 11016145 TI - Printing names of medicines on the strip-sides and using metric units. PMID- 11016146 TI - Adult tetanus immunisation. PMID- 11016147 TI - Cancer of the cervix--problems and challenge of the 3rd millennium. PMID- 11016148 TI - Downstaging of cervical cancer. AB - Globally cervical cancer is the fifth most common cancer and of estimated 460,000 new cases each year three quarters occur in developing countries. In India annually 16% of the world's total cases occur and only 5% are reported in the early stages. Downstaging is defined as a process of screening for cancer using clinical approaches for early detection of this disease. This is distinct from screening test and results in detection of the disease at a less advanced stage in the absence of screening. This experimental approach is applicable in developing countries where cytological screening is not possible in the near future. In this method paramedical staff trained for minimum period will be able to identify any abnormality including suspicious cervix and refer the case early to centres where facilities exist for treatment of premalignant and malignant lesions, including educating the women regarding risk factors, symptoms of the disease and prophylaxis. This experimental methodology recommended by WHO for developing countries like India has to be evaluated by monitoring various ongoing projects where visual inspection screening method is used. The results are collected which include feasibility, compliance, costing, referral methodology, difficulties in implementation, specificity, sensitivity, positive predictive value and drawbacks. The methodology of visual inspection and modified aided visual inspection, frequency and results of various studies in the Indian scenario is for recommendation of downstaging in MCH care. This is to be implemented in rural areas taking into consideration their cultural background and available infrastructure since cytology screening is not possible to cover even 20% of the existing cases in the near future. PMID- 11016149 TI - Radiotherapy in carcinoma cervix. AB - The techniques of radiation treatment of cervical cancer with radium evolved empirically. During the last few decades a lot of technological breakthrough has occurred in the field of brachytherapy mainly due to revolutionary changes brought by computerisation. The difficulties encountered during intracavitary insertions and the practical problems have been discussed. The complications following acute and late reactions have also been discussed. PMID- 11016150 TI - Epidemiology of cancer of the cervix: global and national perspective. AB - Cancer of the uterine cervix is one of the leading causes of cancer death among women worldwide. The estimated new cancer cervix cases per year is 500,000 of which 79% occur in the developing countries. Cancer cervix occupies either the top rank or second among cancers in women in the developing countries, whereas in the affluent countries cancer cervix does not even find a place in the top 5 leading cancers in women. The truncated rate (TR) in the age group 35-64 years in Chennai, India, is even higher (99.1/100,000; 1982-95) than rate reported from Cali, Colombia (77.4/100,000, 1987-91). The cervical cancer burden in India alone is estimated as 100,000 in 2001 AD. The differential pattern of cervical cancer and the wide variation in incidence are possibly related to environmental differences. Aetiologic association and possible risk factors for cervical carcinoma have been extensively studied. The factors are: Sexual and reproductive factors, socio-economic factors (education and income), viruses e.g., herpes simplex virus (HSV), human papillomavirus (HPV), human immunodeficiency virus (HIV) in cervical carcinogenesis and other factors like smoking, diet, oral contraceptives, hormones, etc. The accumulated evidence suggests that cervical cancer is preventable and is highly suitable for primary prevention. Sexual hygiene, use of barrier contraceptives and ritual circumcision can undoubtedly reduce cervical cancer incidence. Education, cervical cancer screening of high risk groups and improvement in socio-economic status can reduce cervical cancer morbidity and mortality significantly. PMID- 11016151 TI - Human papillomavirus (HPV) and cervical cancer. AB - Epidemiologic studies have consistently shown that the most important risk factor for cervical cancer relates to sexual activity and a sexually transmitted agent, probably a virus, is the principal causative agent. In recent years, much attention has been focused on the human papillomavirus (HPV) as the causative agent. The compelling epidemiologic evidence of the role of HPV in cervical cancer is complemented by equally strong data on the role of oncoproteins E6 and E7 of high-risk HPV strains in the molecular pathogenesis of cervical cancer. Incidence and mortality from cervical cancer have been decreasing steadily in most developed countries, but is the leading female cancer in developing countries. This variation in incidence is explicable in terms of differing levels of risk behaviour and population screening facilities and uptake. With better understanding of the aetiopathogenesis, vaccination against HPV is becoming a reality. This may be particularly useful in developing countries, where it is proving difficult to implement effective screening programmes. PMID- 11016152 TI - Immunology of cancer cervix. AB - It is not clear how much genetic factor is responsible for causing cervical cancer, but certainly there has some mutation abnormality in the suppressive p53 gene which is to be proved beyond doubt. In addition to this, there are some environmental factors which make insults on cervical epithelium. Immunology of cancer cervix is well represented by (1) immunology of cancer, (2) immunology of virally controlled antigen and (3) cancer cervix antigen markers. The cancer is an example of transplantation immunology changes in the surface of tumour cells during replication. Human papillomavirus (HPV) DNA has been shown to be associated with high grade cervical intra-epithelial neoplasia (CIN). Squamous cell carcinoma antigen has been investigated for predictive value in cancer cervix. Colony stimulating factor-1 causes monocytic cells aggregation in neoplastic area and plays as initiators of the sequence of inflammation up to neoplasia. PMID- 11016153 TI - An epidemiological survey of carcinoma cervix in north Bengal zone. AB - Correlation between various epidemiological factors and carcinoma cervix patients in North Bengal zone has been studied for the first time. Significant correlation between elderly women (41-50 years age group), low socio-economic status (SES), first coitus before 17 years of age, low literacy rate and this illness has been established. Suggestion has been made for improvement of the picture. Oncology and radiotherapy department of North Bengal Medical College and Hospital should be utilised as the centre for National Cancer Registration Project in North Bengal zone. PMID- 11016154 TI - Diagnosis of carcinoma cervix uteri. AB - Carcinoma cervix should ideally be diagnosed in the pre-invasive or micro invasive stage, or even earlier in the stage of cervical intra-epithelial neoplasia (CIN) by mass screening. As this has not been possible in our country awareness should be created amongst general population and practitioners about the urgent necessity of reporting to a proper clinic as soon as possible after appearance of early symptoms. Downstaging of the cancer should also be performed utilising nurses and paramedical health workers. Early diagnosis would give the best possible results of treatment. PMID- 11016155 TI - Hospital cornea retrieval programme. PMID- 11016156 TI - CME--financial assistance from the government. PMID- 11016158 TI - Medical menace of the millennium. PMID- 11016157 TI - Efficacy and safety of Ascoril expectorant and other cough formula in the treatment of cough management in paediatric and adult patients--a randomised double-blind comparative trial. AB - The present study was undertaken to study the comparative safety and efficacy of two cough formulas viz, Ascoril expectorant and other cough formula in the management of cough associated with respiratory disorders. Fifty patients having cough associated with various respiratory disorders like bronchitis and upper or lower respiratory tract infections were randomly divided into 2 equal groups and were treated with one of the two cough formulas viz, Ascoril cough formula and other cough formula in double-blind manner over a period of 15 days. The evaluation of improvement was carried out by a rating scale using three clinical parameters--cough, sputum and breathlessness. The physicians were asked to rate the effectiveness of the therapy and patients were asked to rate the acceptability of therapy using pre-defined operational criteria. It was observed that the improvement and symptom relief was almost immediate, quicker and better in the group receiving Ascoril as compared to other group. On effectiveness parameter, 96% of the physicians rated Ascoril as having either 'very high effectiveness or high effectiveness' as opposed to only 34% of the physicians who rated other cough formula as having 'high' or 'very high effectiveness'. While on parameter of acceptability, 96% of the patients rated acceptability of Ascoril as 'high' or 'good' as opposed to only 24% of the patients who rated other cough formula 'high' or 'good'. The findings of this study suggests that Ascoril cough formula has better efficacy as well as better patient acceptability. Thus, Ascoril cough formula is superior to other cough formula in management of cough associated with respiratory disorders. PMID- 11016159 TI - Tuberculosis--triumphs and tragedies. PMID- 11016160 TI - Immunodiagnosis in bone and joint tuberculosis. AB - Fifty patients of bone and joint tuberculosis (age 15-70 years) and 30 healthy controls were studied for levels of immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), T cells and circulating immune complexes (CICs) in different stages of disease. Levels of IgG, IgA, IgM, T cells and CICs were significantly raised in fresh and healed tuberculosis patients, compared to control. PMID- 11016161 TI - Tuberculosis: the global epidemic. AB - The modern era of tuberculosis(TB) began in the mid 1980s. In 1993, WHO took the unique step of declaring TB to be a world emergency. Despite this intervention it is estimated that deaths from TB will increase from 3 million a year currently to 5 million by the year 2050. There are 4 principal reasons: World population's increase, co-infection with HIV/AIDS, poverty and programme decline. Other causes contributing to the global epidemic are multidrug resistant TB, immigration, and indifference. The practical solution must concentrate on the completed correct treatment of the disease particularly in those who are sputum smear positive. For this reason WHO is vigorously promoting the Directly Observed Therapy Short course (DOTS) campaign. Doctors treating TB should ideally be part of the public health system. They should have access to first class bacteriological services, good quality of drugs and should make sure that the patient receives the drugs under supervision. Though the reasons for increasing TB are multifactorial it is within the capability of the world to re-exert control providing that the political will is present. PMID- 11016162 TI - Tuberculosis and HIV illness. AB - The HIV infection leading to AIDS is considered to be one of the greatest biomedical challenges in the present century. Like all other communicable diseases AIDS is gradually penetrating the underprivileged sections of society in all countries. Nearly 5 million people in India are living with AIDS at present that makes India a single country with highest number of HIV infected people in the world. The hallmark of HIV disease is the reduction of CD4+ T-lymphocytes, the key cells of immunity. The initial phase of the disease may be termed as HIV seroconversion illness. The next phase is termed asymptomatic HIV infection. When the CD4 count falls below 400/microliter, the patient develops early opportunistic infections which may be termed as early symptomatic HIV infection or AIDS related complex (ARC). Mycobacterial infections are also common in these patients and have led to an alarming resurgence of tuberculosis (TB) in many countries. The available information suggests that TB may be the commonest HIV related disease in the world. The clinical presentation of TB in HIV infected individual is influenced by the degree of HIV related immunosuppression. The atypical features like extrapulmonary TB and absence of positive Mantoux test are generally noticed at a fairly advanced stage. The x-ray chest is also an important adjunct to diagnosis of TB in HIV infected individual. The studies on TB and AIDS in the early part of HIV epidemic in developed countries suggest that extrapulmonary TB is more common in co-infected people but when TB infection is considered in all HIV infected persons, extrapulmonary TB is less common than pulmonary TB. The result of treatment in co-infected pulmonary TB cases is almost similar with the cases suffering from TB alone. Treatment failure, however, has been noticed rarely at extrapulmonary sites. PMID- 11016163 TI - Disseminated tuberculosis. AB - Even though effective chemotherapeutic regimens are available for treatment of tuberculosis (TB), disseminated tuberculosis (DT) continues to be a challenge to physicians. DT represents the unchecked haematogenous dissemination of Mycobacterium tuberculosis. It has been reported that 1.3% of all TB cases are classified as DT. Patients with DT present with a febrile illness of 2 to 4 months' duration. Organomegaly affecting liver, spleen and lymph nodes can be seen in DT. The radiological findings of DT associated with HIV infection are similar to those seen in HIV-seronegative patients. The value of high resolution computed tomography in DT is to demonstrate miliary pattern when chest radiographic findings are atypical or even normal. DT patients may have a mild normochromic anaemia with a normal white blood cell count. Leukaemoid reaction can occur in DT, but pancytopenia is rare. Pulmonary function studies show a mild restrictive ventilatory defect with diffusion defect in cases of pulmonary miliary tuberculosis. Treatment of DT is same as that for pulmonary tuberculosis. Replacement treatment with corticosteroids should be given in tuberculosis patients with adrenal insufficiency. The overall mortality from DT is nearly 20%. PMID- 11016164 TI - BCG vaccine--current status. AB - Tuberculosis incidence has made re-emergence due to multidrug resistance of the bacilli and for acquired immunodeficiency syndrome. Of all vaccines BCG is most widely used. It protects children from secondary forms of tuberculosis. Several authors recommend a second dose to boost the waning effect of BCG. The vaccine at present needs to be improved to make it more efficacious. Vaccines which involve the protective immune responses are ideal. PMID- 11016165 TI - Tuberculosis--historical landmarks. AB - Tuberculosis (TB) since time immemorial has inflicted most miseries in mankind. In ancient times TB was called by many names but the modern one comes from the word 'tubercle'. TB as Pott's disease was widely prevalent among Egyptians in 3700-1000 BC. Hippocrates (460-377 BC) recognised symptomatology of TB. The name tuberculosis was first used by Lanneac and Bayle in early 19th century. Robert Koch in 1882 AD discovered tuberculosis bacillus. Calmette and Guerin laid the foundation BCG vaccination. In 1943, chemotherapy began with the advent of streptomycin (SM) followed by PAS in 1946 and then INH in 1951. Short course chemotherapy results were published in mid 1970s. PMID- 11016166 TI - Diagnosis of tuberculosis. AB - Presence of tuberculous infection in the body does not necessarily mean disease. Any diagnostic work up of the disease starts from a high index of clinical suspicion. However, diagnostic modalities include: (a) Isolation of the bacillus; (b) Immunologic tests; (c) Chemical markers; (d) FNAC, bronchoscopy and bronchoalveolar lavage; (e) Amplification systems. There exists controversies and limitations about the disease process even then. PMID- 11016167 TI - Current recommendations of chemotherapy and pharmacology of antituberculosis drugs. AB - Due to presence of natural mutant bacilli, existence of the bacilli in several populations and in various sites, combination of two, three or more drugs is necessary to treat tuberculosis and that too for a long time. Inadequate therapy leads to relapse and treatment failure. The cumulative effect of negligence, mistakes and new threats (e.g., HIV infection) has made the treatment difficult today. The National Tuberculosis Control Programme of 1962 has been modified recently by Revised National Tuberculosis Control Programme. The targets are to detect at least 70% of new smear positive cases and to achieve at least 85% cure rate by Directly Observed Therapy Short course (DOTS). Categorisation of patients and treatment recommendations are discussed along with pharmacology of antituberculosis drugs. PMID- 11016168 TI - Tuberculosis control in India--past, present and future. AB - Tuberculosis (TB) continues to be a major health problem in India. There has been no perceptible change in the epidemiology of TB since the National Sample Survey 1956-57. Detection of cases has been low over the years since the inception of National TB control programme in 1962 due to passive case finding and high drop out rates among sputum positive patients. Shortening the course of chemotherapy with regimens containing bactericidal and sterilising drugs helped in improving the treatment adherence of patients and cutting down the chain of transmission substantially. Further it is advisable to implement properly the directly observed short course treatment (DOTS) as per WHO guidelines to prevent the development of multidrug resistant TB (MDRTB). Proper management of RNTCP and prevention of MDRTB are all the more important in areas where there is high prevalence of HIV/AIDS co-infected with TB. New vaccine development is also a priority area for research. There is an urgent need for health systems research built into the ongoing programme with proper managerial inputs. PMID- 11016169 TI - Menstrual disorders in genital tuberculosis. AB - Study included 120 cases of genital tuberculosis proved by histopathology. Analysis of their menstrual history showed that the common menstrual disorder was oligohypomenorrhoea found in 54.0% cases, menorrhagia in 19.0% cases, and postmenopausal bleeding in 1.6% cases. History of amenorrhoea was present in 14.3% cases. There were 8 cases of secondary amenorrhoea and one case was of primary amenorrhoea. The pelvic examination showed cervical involvement in 43.1% of cases. Tubo-ovarian mass was present in 19.8% cases. Genital tract tuberculosis is a disease of varied symptomatology. A high degree of suspicion and efficient investigation are important for diagnosis. PMID- 11016170 TI - Retroperitoneal cold abscess with tuberculosis of sacro-iliac joint and pubic bone: a case report of unusual presentation of osteo-articular tuberculosis. AB - Tuberculosis involving sacro-iliac joint and pubic bone presenting with massive retroperitoneal abscess is a rare entity. A 29-year-old female presented with history of discharging sinus in the sacrococcygeal region of 2 months duration. Plain x-ray revealed osteolytic lesion in right pubic bone and left sacro-iliac joint. Computed tomography scan revealed massive pus collection in the retroperitoneal region. Pus was drained extraperitoneally. Biopsy of the scraping of the abscess wall showed granulation tissue with foreign body type of giant cell. On follow-up the patient was doing well. PMID- 11016171 TI - Scourge of tuberculosis. PMID- 11016172 TI - Amendment of Consumer Protection Act. PMID- 11016173 TI - Treating allergies: a perspective in family practice. AB - The family practitioner is the first and most important contact person for a patient suffering from any allergic disorder. The reason is simple. Even in this fast-changing world of the twentyfirst century, the family practitioner will continue to be not only a doctor but also a friend, guide and philosopher to many a sufferer in our country. It is therefore important for the primary care physician to take time out for periodic reviews of the therapeutic scenario, which in fact is the objective of this article. PMID- 11016175 TI - Clinical profile of falciparum malaria in a tertiary care hospital. AB - Falciparum malaria presents with protean manifestations and is associated with a variety of complications and has a high mortality. One hundred and fifty-eight consecutive cases of falciparum malaria were studied with respect to the clinical presentation, complications, and response to treatment. The mean age of patients was 38.60 +/- 15.45 years and majority of them were males i.e., males being 110 (69.62%) and females being 48 (30.37%). The commonest presenting manifestations were fever with chill and rigor (98.10%), altered sensorium (48.10%), algid malaria (18.35%), and jaundice (27.21%). The other presenting features being oliguria (6.96%) and bleeding manifestations due to disseminated intravascular coagulation (DIC) (4.43%). The frequently encountered complications were anaemia (74.68%), jaundice (40.50%), cerebral malaria (45.56%), thrombocytopenia (40.50%) and renal failure (24.68%). Most of the patients i.e., 126 (79.74%) recovered with treatment and 32 (20.25%) succumbed. Higher mortality was associated with higher parasite count, presence of complications like anaemia, jaundice, renal failure, DIC, adult respiratory distress syndrome (ARDS), and septicaemia. Most of the deaths were encountered in patients where there was delay in clinical diagnosis, in the pre-hospital phase, and consequent presentation in multiorgan failure. Early diagnosis and institution of specific therapy were rewarding in the remaining patients in this series. PMID- 11016174 TI - Idiopathic intracranial hypertension. PMID- 11016176 TI - Evaluation of women seeking sterilisation reversal. AB - A total of 1120 women attending at Regional Centre of Excellence (RCOE), RG Kar Medical College and Hospital, Calcutta from October, 1989 to July, 1998 with the request for sterilisation reversal (SR) were evaluated step by step for fitness of SR operation on the basis of history, clinical examination, pre-operative investigations, fertility status and laparoscopic findings. One thousand and sixty couples completed their evaluation. Majority of the SR seekers were young (< 30 years) and significant number (8.9%) of women came in late age (> 40 years). Most (85.8%) of the women had one or no child. The main reason for requesting reversal was death of children (91.8%). Only 43.3% were ultimately found fit for SR operation and rest (56.7%) were refused. Presence of 2 or more children in not remarried couple (13.8%), gross pelvic pathology (2.35%), advanced age (8.7%), seminopathy (4.05%), and some medical diseases (3.2%) were important reasons before pre-operative investigations for refusal. Faulty sterilisation technique (11%) was responsible for refusal at laparoscopic evaluation. The need of critical evaluation of all women seeking reversal has been emphasised in this study. Various aspects related with demand for SR and preventive measures to improve the success of this procedure have been critically discussed. PMID- 11016177 TI - Selenium levels in dilated cardiomyopathy. AB - Thirty-seven dilated cardiomyopathy cases have been studied and compared with 20 normal controls. Serum selenium levels in relation to coronary risk factors were studied. Serum samples were analysed for selenium, total cholesterol, high density lipoprotein(HDL) cholesterol, triglycerides and glucose levels. Smoking, alcohol intake, positive family history, psychosocial tension, obesity, hypercholesterolaemia, hypertriglyceridaemia and hyperglycaemia were found in the following percentages 27%, 8%, 5%, 73%, 41%, 38%, 81%, 46% respectively in dilated cardiomyopathy patients. Low selenium (< 4.5 micrograms/dl) and HDL cholesterol levels and high total cholesterol, triglycerides, low density lipoprotein cholesterol, very low density lipoprotein cholesterol and glucose levels were observed in dilated cardiomyopathy cases compared to controls. The present results support the concept that low selenium levels along with other risk factors play an important role in developing dilated cardiomyopathy. PMID- 11016178 TI - Seroprevalence of brucellosis in Kashmir (India) among patients with pyrexia of unknown origin. AB - Over a period of 5 years from 1992 to 1997 a total of 3,532 patients of pyrexia of unknown origin (PUO) were subjected to Wright's tube agglutination test for brucellosis. Of the 3,532 patients tested, 28 (0.8%) were found seropositive for brucellosis. Males outnumbered females by a ratio of 3:1. Whereas the seroprevalence among males clearly was age-related, the seroprevalence among women was not. The highest number (43%) of positive males belonged to the age group 21-30 years. The majority of patients had titres of 1:160 or 1:256, high titres of 1:512 and 1:1024 were found in 21.4% patients. PMID- 11016179 TI - Role of cryopreservation in the treatment of male subfertility. AB - A comparative study of routine seminal parameters of fresh and frozen thawed semen was carried out among 90 samples. Three classes of semen viz, oligozoospermic, asthenozoospermic and normozoospermic samples were investigated in both the cases for survival rate, per cent motility and viability of the cells. Regarding post-thaw motility the samples exhibited mixed reaction. In all other cases a class specific response was observed. PMID- 11016180 TI - Evaluation of peritoneal lavage and abdominal paracentesis in cases of blunt abdominal trauma--a study of fifty cases. AB - Diagnostic peritoneal lavage (DPL) is an accurate technique to diagnose abdominal visceral injury in blunt abdominal trauma. Four-quadrant abdominal paracentesis should be used routinely in all cases of blunt abdominal injuries as a diagnostic tool. The study included 50 cases in whom four-quadrant abdominal paracentesis was done at JN Medical College, Aligarh. There were 13 positive taps, out of which 12 were true positive and one was false positive. In all 37 negative taps DPL was done, out of which 23 had positive lavage, 22 had true positive and one had false negative results. All the patients with true positive peritoneal lavage had intra-abdominal injuries at laparotomy. PMID- 11016181 TI - The problems of hypertension in the elderly. AB - The cut off age for elderly person in India is 60-65 years, in the USA is 75-80 years and 6th Joint Committee on Detection, Evaluation and Treatment of High Blood Pressure (JNC-VI) has identified it as above 60 years. Elderly people may have (i) systolic-diastolic hypertension, (ii) isolated systolic hypertension or (iii) pseudohypertension. JNC-VI has classified hypertension in stage 1, stage 2 and stage 3 according to its severity. Hypertension is confirmed when BP measured on three separate occasions over 1-2 weeks and when consistently it is raised above 140/90 mm Hg. The management includes lifestyle modification and drug treatment. Lifestyle modification includes rationality of diet, regular exercise, stop smoking, stoppage of alcohol or moderation and yoga. Drugs commonly used are diureties and beta-blockers. Other antihypertensive drugs are calcium channel blockers, ACE-inhibitors, alpha-blockers and vasodilators. PMID- 11016182 TI - Recent advances in the management of Alzheimer's disease. AB - Alzheimer's disease is a chronic degenerative disorder of the central nervous system. As the disease progresses there is gradual increase of mental and physical incapacitation, ultimately leading to death. Recent advances in the management of the disease focused the most extensive drug development activity towards the anticholinesterase agents. PMID- 11016183 TI - Posterior urethral valves with congenital megalo-urethra. AB - A 3-year-old male child was admitted with complaints of dysuria and dribbling of urine since birth. There was a small plum sized swelling, on the ventral surface of penis, which increased in size during micturition and on compression of the swelling urine dribbled out. Voiding cysto-urethrogram showed posterior urethral valves with scaphoid megalo-urethra. Vesicostomy with fulguration of the valves and one year later reduction urethroplasty were performed. PMID- 11016184 TI - Intestinal perforation--a unique cause. AB - An illiterate, 65 years old, male attended surgery emergency with features of perforation peritonitis. Exploratory laparotomy revealed perforation in the terminal ileum and a sharp metallic object pointing at the perforation site. This, on removal, was found to be the sharp edge of a blister pack (with intact tablet within). The perforation was repaired. The abdomen was closed after leaving a drain. Postoperatively the patient had a morbid period and ultimately responded to management. PMID- 11016185 TI - Digital soft tissue Ewing's sarcoma of foot--a case report. AB - A 22-year-old female presented with rapidly increasing, fungating and painful growth of right foot of 4 months duration. The growth had superadded infection. Radiography revealed soft issue mass between 2nd and 3rd toes without any bony involvement. Histopathologically it was confirmed to be a case of extraskeletal Ewing's sarcoma. Amputation above the ankle was done. She was advised for chemotherapy. She is still alive after 2 years. Here in this case the tumour arose in the subcutaneous tissue between 2nd and 3rd toes and diagnosed as a case of Ewing's sarcoma of the foot. PMID- 11016186 TI - An unusual case of double outlet right ventricle. AB - A 17-year-old unmarried girl was admitted with the complaints of breathlessness on exertion, swelling of upper and lower limbs and intermittent fever for last 21 days. Past history revealed bluish discolouration of nails and tongue since birth along with recurrent dyspnoea. After thorough general and cardiovascular examination, a provisional diagnosis of tetralogy of Fallot was made. Chest x-ray revealed cardiomegaly and absent main pulmonary artery segment. ECG revealed gross right axis deviation and right ventricular hypertrophy. Echocardiogram showed features of double outlet right ventricle. PMID- 11016188 TI - Blood donation reduces elevated blood lipids. PMID- 11016187 TI - Stromal sarcoma of breast: a case report. AB - A 34 years unmarried female was admitted with an ulcerated foul smelling growth in her right breast. On examination the fungating mass measured 17.5 cm x 15 cm in central and lower part of right breast involving the nipple and areola. The ulcer was covered with slough and rest part of the breast appeared bosselated. Her Hb was 4 g/dl and incision biopsy from the margin of the tumour showed histology of sarcoma. The patient was infused 6 units of blood and right sided total mastectomy was done. Histopathological examination confirmed it was a case of stromal sarcoma of breast. Chemotherapy was started with vincristine, adriamycin and cylophosphamide. The patient was doing well in next follow-up. PMID- 11016189 TI - Problem in injecting penicillin. PMID- 11016190 TI - Pulse polio Immunisation Programme (PPIP) PMID- 11016191 TI - Evaluation of safety and efficacy of supirocin-B (mupirocin 2% + betamethasone dipropionate 0.05%) in infected dermatoses--a post marketing study. AB - The aim of the present post marketing study was to study the safety and efficacy of supirocin-B ointment (mupirocin 2% + betamethasone dipropionate 0.05%) in the treatment of infected dermatoses. For this purpose physicians from different parts of India were requested to keep the clinical records prospectively as per a specially designed proforma over a follow-up period of 7 days, whenever they prescribed supirocin-B ointment (mupirocin 2% + betamethasone dipropionate 0.05%) for local application, three times a day, to their patients having either primary infection complicated by dermatoses or dermatoses infected secondarily. From the analysis of 251 clinical records contributed by 27 physicians, it was evident that in clinical practice, supirocin-B ointment (mupirocin 2% + betamethasone dipropionate 0.05%) was found to be safe and very effective by physicians in the treatment of infected dermatoses in 94.8% of the patients. Similarly 92.4% of the patients reported more than 70% improvement in their symptoms after 7 days of treatment. No adverse effects were reported during the treatment period by any of the patients except worsening of skin lesions by one patient. Thus from this study, supirocin-B ointment (mupirocin 2% + betamethasone dipropionate 0.05%) seems to be safe and effective in the treatment of infected dermatoses. PMID- 11016193 TI - Safe blood starts with me: blood saves lives. PMID- 11016192 TI - Spectrum of antibiogram against pathogens related to respiratory tract infections with a special reference to ceftibuten: a multicentric study. AB - In a multicentric study at several leading hospitals of this country, microbiological assessment was carried out in 500 specimens from patients suffering from respiratory tract infections (RTIs; both upper and lower) for a period of 6 months from January, 1999 to June, 1999. The antibiotic sensitivity study was done in 201 isolates from 500 different specimens of throat swab, postpharyngeal swab, sinusitis drainage fluid, sputum, broncho-alveolar lavage (BL), etc. Ceftibuten, an orally active third generation cephalosporin showed encouraging results when compared with seven other selected antibiotics used for RTI. The majority of the patients with acute or chronic RTIs showed an excellent in vitro response to ceftibuten in the analysis of the isolates. Seventy to ninety per cent of the isolated respiratory pathogens were found to be sensitive to ceftibuten in vitro; which offers a promising alternative to other antibiotics included in this study. PMID- 11016194 TI - [Development in cardiovascular genetics. Drugs adapted to patient's genetic profile soon available]. PMID- 11016195 TI - [World champion in ECLS/ECMO? Large material from the ECLS clinic in Michigan presented in JAMA]. PMID- 11016196 TI - [Microglia--new target cells for neurological therapy]. AB - Disturbances in the normal homeostasis of the central nervous system induce a localized activation of microglia. This activation serves to isolate pathological processes from surrounding, intact nervous tissue. Concomitantly, healthy or minimally damaged nerve cells nearby may be negatively influenced by potent molecules released by activated microglia. This situation appears to exist e.g. in ischemia, multiple sclerosis and Alzheimer's disease. Pharmacologic regulation of microglial activity is therefore a rational approach to treatment of many central nervous system disorders. PMID- 11016197 TI - [Improved survival after in-hospital cardiac arrest]. AB - Internationally, survival among patients suffering in-hospital cardiac arrest is relatively low and unchanged at about 15%. Our experience at Sahlgrenska Hospital in Goteborg indicates a higher rate. We found survival to be related to the type of arrhythmia initially encountered, the highest rate having been observed among patients in ventricular fibrillation. We also found survival among patients suffering cardiac arrest to be higher in monitored as opposed to non-monitored wards. Whether the improved survival rate observed at Sahlgrenska as compared with international observations among patients suffering cardiac arrest is due to improvements in the organization or to patient selection is not clear. In order to achieve a higher survival rate after in-hospital cardiac arrest an efficient organization in which health care providers are given regular training and feed back is most likely of the utmost importance. PMID- 11016199 TI - [Reminders for lung auscultation and percussion]. PMID- 11016198 TI - [Dramatic reduction of myocardial infarction mortality. A retrospect of the past century]. AB - Ischemic heart disease is the most common cause of death in the western world. Acute myocardial infarction is one of the most life-threatening manifestations of ischemic heart disease. Treatment of acute myocardial infarction has dramatically improved over the past 100 years by the introduction of coronary care units, mobile coronary care units, improved treatment of cardiac arrest plus the introduction of beta-blockers, fibrinolytic agents, aspirin, ACE-inhibitors and lipid lowering drugs. This has resulted in a marked reduction of mortality. PMID- 11016200 TI - [Medical revision a way to treat blindness when it comes to home defects. Report from the first scrutiny of psychiatric care]. AB - Results from medical revision of eight psychiatric clinics show that the points of view of "outsiders" can be of help to improve routines and procedures. PMID- 11016201 TI - [Traditional quality assurance is not enough! Focusing on the patient demands modern, learning-based development]. PMID- 11016202 TI - [New strategies for the Swedish classical field of salmonella control]. PMID- 11016203 TI - [Information is required to increase the number of organ donors]. PMID- 11016204 TI - [The Lasker Prize 1999. Molecular structure of ion channels as a theme of awarded research]. PMID- 11016205 TI - [The Swedish Medical Society and the sterilization law in 1929-1974]. PMID- 11016206 TI - [The unknown history of lobotomy: women, children and idiots were lobotomized]. AB - The history of prefrontal lobotomy is an interesting example of medicine regarding as useful a treatment method which present-day consensus evaluates in a contrary fashion. A pilot study of archives from the Swedish state mental hospital Umedalen shows that the frequency of lobotomies as well as postoperative mortality were higher than what has earlier been assumed. The majority of the 704 patients who underwent lobotomy at Umedalen hospital were women. One unexpected finding concerns the numbers of mentally retarded patients and children who were subjected to lobotomy. Case records and other documents from the hospital archives indicate that the operation was performed largely for the benefit of the hospital rather than the patient, with an eye to engendering calm and order on the unruly wards. PMID- 11016207 TI - [Circumcision--a Jewish reply: "Hofvander and Olsen" act as enlightened European cultural imperialists"]. PMID- 11016208 TI - [A reply: rights of the children are more important than parents' right to the circumcision of their children]. PMID- 11016209 TI - [Where is the Lakartidningen going? Critical views on the series on scientific theory: Raise elementary demands on lucidity!]. PMID- 11016210 TI - [Reply 1: Qualitative surveys are not based on ruling hypotheses]. PMID- 11016211 TI - [Reply 2: It's the platform contact, not the observation which constitutes the discovery]. PMID- 11016212 TI - [Alfred Szamosi comments: The articles on the scientific theory bring unfortunately no guidance]. PMID- 11016213 TI - [Should homosexuals be allowed to adopt children? What is the opinion of the Swedish Medical Society?]. PMID- 11016214 TI - [Uncritically about Cox-2 inhibitors]. PMID- 11016215 TI - [A reply: Not without costs to prevent hundreds of NSAID-related fatal cases]. PMID- 11016216 TI - [Concerning guidelines on care of brain injuries: a national randomized trial is planned]. PMID- 11016217 TI - [Quality assurance is safety work!]. PMID- 11016218 TI - [The National Board of Health and Welfare on sexual abuse of children: incorrect statements on the administration of justice]. PMID- 11016219 TI - [Comments on heart failure: resource utilization was accounted for]. PMID- 11016220 TI - [Quality assurance--weapon safety--discharged. As Luther we can ask: "What is it?"]. PMID- 11016221 TI - [Four surgeons introduced ether inhalation in Sweden. Then the anesthesiology was put to sleep for 100 years]. PMID- 11016222 TI - [Impossible to document adverse effects of calcium antagonists. Effects equal to those of conservative treatment of hypertension]. PMID- 11016223 TI - [A comment on calcium antagonists: what has been elucidated?]. PMID- 11016224 TI - [Minidrainage in pneumothorax is expensive, but still beneficial. Considerate for the patient, reducing the number of hospitalization days]. AB - Initial tests (ten patients) are reported of a compact device, Tru-close, for the evacuation of pneumothorax. It consists of a small-bore plastic catheter combined with a small box containing a flutter-valve. Ease of insertion, safe function, stable fixation and painless removal were features found. Complications were pain on insertion in one patient and leakage of pleural fluid from the device in another. The unit is expensive, but in cases of simple pneumothorax without fluid it would seem possible to manage patients on an out-patient basis, thus saving several days' worth of hospitalization costs. PMID- 11016225 TI - [Excision and suture in skin and subcutaneous tissue. Basic techniques of plastic surgery are essential for optimal cosmetic results]. AB - The basic techniques of plastic surgery are of great practical importance in achieving the best possible results when reconstructing skin and subcutaneous tissue. Knowledge of the anatomy of the skin and the normal healing process is also essential. These fundamental principles of skin surgery include appropriate methods of infiltration anesthesia. Detailed descriptions of the technique of skin incision, excision of skin lesions and control of bleeding are given. Techniques of skin suture are described, and recommendations for optimal timing of suture removal and treatment of scar tissue are provided. PMID- 11016226 TI - [WHO spearheads global initiative to eradicate hepatitis B]. AB - It is estimated that over 350 million people live with a chronic hepatitis B virus (HBV) infection, claiming over one million deaths per year due to progress of the chronic disease to cirrhosis and/or hepatocellular carcinoma (HCC). An extended program of immunization including hepatitis B vaccine for children under one year of age has been launched in more than 110 countries. Recent studies conclude that mass hepatitis B immunization is effective in preventing HBV infection and has resulted in a decrease in the occurrence of HCC in children living in countries where hepatitis B is endemic. However, the vast majority of infected children live in the poorest developing countries in Africa and Asia that currently cannot afford the vaccine or lack the basic infrastructure necessary to deliver a national immunization service. The Global Alliance for Vaccines and Immunization (GAVI) was established in 1999 as an alliance of WHO, UNICEF, the World Bank, industry, foundations, and other partners to reinvent immunization for the 21st century, by forging a common vision and new ways of working together at global, regional and national levels. WHO recommends global elimination of hepatitis B by universal infant and/or adolescent immunization, but health planners in Sweden and the other Scandinavian countries, the Netherlands and UK have not yet been convinced of the cost-effectiveness of HB prevention through routine childhood immunization with HB-vaccine. The inclusion of hepatitis B vaccine in already available multivalent vaccines may alter this situation in the future, but until then an intensified vaccination strategy aimed at those groups of individuals that are particularly at risk for hepatitis B should be adopted in accordance with the recommendations of The Swedish National Board of Health (SOSFS 1991:2) and local instructions from the County Medical Officer for Communicable Disease Control. PMID- 11016227 TI - [A case report. Surgery for incontinence caused bacteriuria and bladder calculi]. PMID- 11016228 TI - [Medical education--developing or blunting? Students don't receive support from teachers according to a questionnaire study]. PMID- 11016229 TI - [Learning among medical students. Teachers' commitment a decisive factor]. AB - During their medical education, students take many courses in a variety of scientific subjects. This is most noticeable during the 5th term, when 6 subjects are taught in parallel during a 10 week period. This makes the process of learning very important. Using a modified written questionnaire we asked for students' opinions on factors important for learning. For crucial parameters such as coherence in knowledge and study motivation the students considered teachers' interest in students' learning to be the critical factor. PMID- 11016230 TI - [Interpreting messages from across knowledge boundaries in health care encounters]. AB - Anthropological research into health issues may contribute to the discussion regarding doctor/patient communication and to understanding across knowledge boundaries. Anthropological studies in "other cultures" provide a mirror, and the question "Why are we doing what we are doing?" becomes relevant. Examples from research among immigrants in Swedish health care bring this question into focus. The relationship between scientific knowledge and lay knowledge is seen as problematical with regard not only to immigrants but also to any patient involved in today's preventive care, steeped as it is in rapid medical technological development. Scientific knowledge is recontextualized, interpreted and understood in all clinical situations. Research into this process is an urgent matter. PMID- 11016231 TI - [Conflicting values in evaluation of psychiatric care--a review]. AB - Greater understanding is needed regarding psychiatric disorders and their causes, as well as the effects of psychiatric care. Ethical issues ought to be discussed in terms of both the choice of questions and to the ways in which data are collected. So far, most evaluations of psychiatric care have focused on medical utility, while issues of autonomy and impartiality are sorely lacking. An important challenge for the future is therefore to support the evolution of the search for knowledge into a multidisciplinary activity in which values concerning autonomy and impartiality are also incorporated. PMID- 11016232 TI - [Geriatric psychiatry in an international perspective. Three important WHO documents now in Swedish translation]. PMID- 11016233 TI - [The lopsidedness of psychiatry. A historical, but subjective perspective]. PMID- 11016235 TI - [To be able or not to be able--that is the primary question!]. PMID- 11016234 TI - [Medical historical pause. Physician Bill in Lappland as a challenge for two painters]. PMID- 11016236 TI - [Deficient routines in management of Creutzfeldt-Jakob disease]. PMID- 11016237 TI - [Screening for cervix cancer--a successful program that should be improved]. PMID- 11016238 TI - [Does Sweden need emergency medicine specialists? Yes, but there are conditions]. PMID- 11016240 TI - [Medical records via Internet? Use the technology]. PMID- 11016239 TI - [Role of primary health care--a (critical) scrutiny of the report by the National Board of Health and Welfare]. PMID- 11016242 TI - [Organ donation and respect to death]. PMID- 11016241 TI - [Life as a gift--a group for public information on organ donation]. PMID- 11016243 TI - Drafting. PMID- 11016244 TI - Optometrists in the mainstream--2010 and 2020. PMID- 11016245 TI - Meditation as therapy. PMID- 11016246 TI - Healthy People 2010--vision objectives for the nation. AB - BACKGROUND: In January 2000, the U.S. Department of Health and Human Services released new national health goals and objectives in a report entitled Healthy People 2010. This national report includes specific objectives for improving the vision of people in the United States during the 2000-2010 decade through prevention, early detection, treatment, and rehabilitation. The vision objectives, along with other health objectives, are designed to help the nation achieve two major outcomes: (1) increase the quality and years of healthy life and (2) eliminate health disparities among different groups. These health objectives will strongly influence many health care programs and the provision of health care through laws, regulations, reimbursements, and clinical guidelines enacted by federal and state agencies. PURPOSE: The purpose of this article is to inform optometrists about the new Healthy People 2010 vision objectives. METHODS: A summary of the Healthy People process for developing the objectives--and the specific vision objectives--is provided. These objectives address many important areas, including: regular dilated eye examinations; vision screening for preschool children; uncorrected visual impairment due to refractive errors; blindness and visual impairment in children and adolescents; visual impairment due to diabetic retinopathy, glaucoma, and cataracts; occupational eye injury; the use of personal protective eye wear in recreational activities and hazardous situations around the home; and the use of vision rehabilitation services and adaptive devices by people with visual impairments. CONCLUSION: These new national health goals and objectives will impact optometrists. Therefore, it is important that optometrists understand these objectives and actively work to help implement them at the national, state, and local levels. By doing this, they can help provide the services needed to reduce visual impairments and improve the vision of people in their communities. PMID- 11016247 TI - Test-retest reliability of the College of Optometrists in Vision Development Quality of Life Outcomes Assessment. AB - BACKGROUND: Medical diagnoses such as headaches, diplopia, nausea, and asthenopia are conditions that are often treated with optometric vision therapy. Improvement of such subjective conditions after a regimen of therapy is often difficult to objectively quantify. The goal of this study was to verify the test-re-test reliability of a new clinical survey instrument, the College of Optometrists in Vision Development (COVD) Quality of Life Outcomes Assessment, which allows the analysis of symptoms data. METHODS: The first-year class of optometry students at the Oklahoma College of Optometry at Northeastern State University were administered the instrument in a group setting on two different occasions, two weeks apart. Nineteen students completed both test and re-test. Statistical analysis by the Wilcoxon Signed Rank Test and paired t test were performed. RESULTS: Each subject reported some of the 30 symptoms on both test and re-test. The instrument was found to be reliable. Wilcoxon Signed Rank Analysis showed no significant differences in test-retest scores, either pooled or item-by-item. A paired t-test group and item analysis were insignificantly different between scores. Spearman's rho correlation for test-retest of each subject was 0.878. Eighty-nine percent (17 of 19) scored insignificantly different between administrations. Ninety percent of the items (27 of 30) were found to vary insignificantly between the two administrations. CONCLUSIONS: The COVD Quality of Life Outcomes Assessment is a reliable tool to measure changes in symptoms on the basis of optometric intervention--specifically, vision therapy. PMID- 11016248 TI - Pseudoduplication of the optic nerve head. AB - BACKGROUND: Pseudo-doubling of the optic nerve head is a spectacular clinical entity, in which a lesion resembling an optic disk appears adjacent to the true optic disk. CASE REPORT: A case of unilateral pseudo-doubling of the optic disk with bilateral optic nerve pits is presented. CONCLUSIONS: The lesion is congenital, and represents a chorioretinal coloboma with optic disk involvement. Pseudo-doubling can be differentiated from true doubling of the optic nerve by the imaging techniques of ultrasonography, computerized tomography, and magnetic resonance imaging. PMID- 11016249 TI - Cytomegalovirus retinitis: pathogenesis and treatment options. AB - BACKGROUND: While patients infected with the human immunodeficiency virus (HIV) are displaying less cytomegalovirus (CMV) retinitis as a result of the use of a combination of anti-HIV medications, it still remains a serious threat to vision in this population. METHOD: This review examines the nature of CMV infection, in general, and discusses the pathogenesis, clinical features, and treatment options for CMV retinitis, in particular. PMID- 11016250 TI - Practice strategies. Regional variations. PMID- 11016251 TI - Enriching staff positions. PMID- 11016252 TI - Cash basis accounting. PMID- 11016253 TI - Subsequent care codes appropriate for nursing home care. PMID- 11016254 TI - [The tuberculosis incidence in Romania in the 1st quarter of 2000]. PMID- 11016255 TI - [The prevalence of smoking in physicians in Romania]. AB - An inquiry upon the prevalence of the habit of smoking in doctors in Romania, performed in a group of 1136 doctors (p = 0.05) revealed a prevalence of 43.2%, 50.1% in men and 38.6% in women. Smoking is more frequent in group age 30-49 (47.2%). More than half of the smoking doctors smoke between 10 and 20 cigarettes a day. 45.7% of the resident doctors are smokers. Taking into account the large interface with the public, the medical staff can play an important part in the activity of reducing or stopping smoking. PMID- 11016256 TI - [The prevalence of bronchial asthma in 13- to 14-year-old schoolchildren in the city of Bistrita]. AB - Asthma is a major worldwide public health problem, with a prevalence in children which varies in different geographical areas from 0 to 30%. The true prevalence of the disease is not known exactly due to the lack of consensus of its definition and because the different methods used by the studies for obtaining epidemiological data. The authors study the prevalence of asthma among schoolchildren aged 13-14 years, from Bistrita, using the ISAAC standardised methodology, world-wide accepted. A total number of 889 schoolchildren aged 13-14 years from local schools, were enrolled in the study, the sample representing 63.67% of all 1396 schoolchildren of the same age registered in the 1996/1997 school year in Bistrita. The first step consisted of completing a questionnaire about asthma, allergic rhinitis and eczema. Seventy-five (8.43%) schoolchildren gave positive response to asthma and asthma associated symptoms, with a prevalence of 7.43% in boys (33 from the total of 444) and 9.43% in girls (42 of 445). In the second step 61 schoolchildren of those who gave positive responses to the questionnaires about asthma-wheezing, representing 81.33% of the total number of 75, were complex examined (history and physical examination, respiratory function and allergological tests). The following results were obtained by correlation of questionnaires and complex examinations: 17 subjects known with asthma before completing the questionnaire, 1 with probable asthma and 21 with certain asthma. Those 39 children represent 4.38% of the total number of schoolchildren who had been examined, with a gender rate of 19 boys (4.27%) and 20 girls (4.49%). In conclusion, the estimated asthma prevalence among 13-14 years old schoolchildren from Bistrita is 4.38%, with a similar gender rate. The prevalence data found for 13-14 years old schoolchildren in Bistrita are comparable to those made in other geographical areas in Romania, and to those reported by former socialist countries from Eastern Europe. PMID- 11016257 TI - [CPAP--a still current therapy?]. PMID- 11016258 TI - ["Pseudomalignant" lung tumors--a permanent diagnostic and treatment challenge]. AB - A clinical case of dysembryoplasic benign tumor, respectively a lung hamartoma, is presented. In this case, the pre-operational diagnosis was malignant tumor at the operationally limit, but the histopathological examination of the operational specimen was of lung hamartoma. PMID- 11016259 TI - [Therapeutic pneumothorax--an effective adjuvant method in treating multidrug resistant tuberculosis]. AB - Treatment of multidrug-resistant TB is difficult, even when all alternative drugs are at hand, due to important side effects and high cost, buying seldom efficient. Therapeutic pneumothorax (TP) proved its efficacy in pre-antibiotic era, so one can assume that it can be applied with the same success in multidrug resistant TB. Three MDR TB cases are presented (resistance to 2, 3 and 4 drugs), with unilateral lesions, in which TP was applied as a help for the drug treatment which was guided by the antibiogram. Favorable results were obtained in all 3 cases, with constant bacteriological negativity. In one of the cases (with 4 drugs resistance), treatment was completed after 3 months by a lobectomy; they were all considered healed after 18 months. In conclusion, in some cases of MDR TB, the TP (if there are no pleural adherences) may lead to good results. This presentation aims to remind the pulmonologists the favourable effects of TP in selected cases. PMID- 11016260 TI - [Tracheal reconstruction via a homograft--from an experiment to clinical applications]. PMID- 11016261 TI - [The possibilities and limits of bacteriologic diagnosis in nontuberculous bacterial lower respiratory infections]. PMID- 11016262 TI - [A fatal prognosis and the physician's dilemmas]. PMID- 11016263 TI - [What is your diagnosis? Secondary to chest trauma, the patient developed hemothorax]. PMID- 11016264 TI - [Policy and advocacy, 2 words that cannot be translated]. PMID- 11016265 TI - [Immunosuppressive mechanisms for immunoglobulin function]. AB - The article reviews the evidence supporting and explaining suppressive activity of intravenous immunoglobulin (IVIG) preparations towards human immune system. Besides basic mechanisms also clinical applications and effects in various autoimmune disorders such as idiopathic thrombocytopenia, Guillain-Barre syndrome and others are described. This evidence supports novel application of IVIG as convenient alternative to other immunosuppressive therapies with many more adverse effects. PMID- 11016266 TI - [Immunomodulation of pain]. AB - Review of the interacting elements of nociceptive, neuroimmune and neuroendocrine systems with a stress on sympathetic innervation of the lymphoid organs. Special attention is given to dysfunctions within the nociceptive, neuroendocrine and immune systems due to: damage to the: 1) afferent fibers; 2) sympatho-adrenal system; 3) structure of the hypothalamo-pituitary-adrenal axis. The role of peripheral immunocytes in the generation of pain, including local cytokines, opioids and CRF is also discussed. PMID- 11016267 TI - [Cardiotoxicity of anthracycline]. AB - The anthracyclines are among the most active and useful anticancer agents, but they are also the most common chemotherapeutics associated with the development of congestive heart failure. Twenty seven to sixty percent of patients with anthracycline-induced cardiomyopathy die of congestive cardiac failure. The paper presents the pathomechanisms of anthracycline-induced cardiotoxicity and perspectives on prevention of congestive heart failure resulting from anthracycline chemo-therapy. PMID- 11016268 TI - [Epidemiology, pathogenicity and diagnosis of echinococcosis]. AB - Echinococcosis is an infectious disease caused by the larval form of the genus Echinococcus. Two of the parasite species may cause severe disease in humans: Echinococcus granulosus and Echinococcus multilocularis. The liver is the most frequently involved organ in patients with echinococcosis. Diagnosis is achieved by variety of clinical methods, such as computerized tomography, ultrasound examination, magnetic resonance imaging and immunodiagnosis based on the detection of patient's serum antibodies. PMID- 11016269 TI - [Molecular markers of ovarian carcinoma]. AB - Definition of a tumor marker and significance of proper choice of ovarian cancer marker in diagnosis, prognosis and monitoring effects of chemotherapy, are pointed out. Markers of malignant ovarian tumors were classified into oncogenes and oncoproteins, antigens, hormones, enzymes and their inhibitors, acute phase proteins (cytokines, growth factors, haptoglobin), hemostasis and some others. Methods of quantitative determinations of cancer markers are presented. PMID- 11016270 TI - [Renin-angiotensin-aldosterone system in thyroid disorders]. AB - Alterations of the renin-angiotensin-aldosterone system activity occur in patients with thyroid disfunction. This paper is a review of the effect of hyperthyroidism or hypothyroidism in human beings and experimental animals on changes of the renin, angiotensinogen, angiotensin converting enzyme, angiotensin II and aldosterone concentrations and plasma renin activity. PMID- 11016271 TI - [Mechanisms for development of tolerance to repeated injections of exogenous pyrogens]. AB - Pyrogenic tolerance has been recognized for many years in a variety of species although the mechanisms that are responsible for its development are not well understood. The development of pyrogenic tolerance is associated with the stepwise diminution of pathophysiological and behavioral responses induced by exogenous pyrogens, such as fever, reduction in food and water intake. Several studies either in vivo or ex vivo have indicated the role of various proinflammatory cytokines in the development of pyrogenic tolerance. Most of these studies have indicated that pyrogenic tolerance is associated with down regulation of cytokine production as well as their biological activity. The mechanisms responsible for down-regulation of cytokine production during development of pyrogenic tolerance are unclear. Since glucocorticoids are required for induction of tolerance, it has been postulated that well known glucocorticoids-dependent negative feedback on the production and biological activity of cytokines may play an important role in development of pyrogenic tolerance. We can not, however, rule out possibility that other mechanisms may participate directly or indirectly in a suppression of cytokines response due to repeated exogenous pyrogen challenge. Either the enhanced uptake of exogenous pyrogens by the hepatic Kupffer cells or the desensitization to exogenous pyrogens by the loss of binding sites, have been proposed as an additional mechanisms which may participate in exogenous pyrogen hyporesponsiveness. PMID- 11016272 TI - [Automatic analysis of reticulocytes--new diagnostic possibilities in hematology]. AB - This review describes a new method of reticulocyte analysis based on flow cytometry, better than manual reticulocyte counting. Reticulocyte percentage, absolute number, indices of maturity (RMI--reticulocyte maturity index, IRF- immature reticulocyte fraction) and others qualitative parameters express the erythropoietic activity of kidneys and bone marrow. The new proposed parameters can improve the diagnosis of anemias and others haematological disorders like leukemia, myelodysplastic syndrome. They also provide exactly and early information about the status of bone marrow after ablative chemotherapy and transplantation. Automated reticulocyte counting can create new potentials for haematology diagnosis. PMID- 11016273 TI - A new approach to the diagnosis of EHEC. PMID- 11016274 TI - Non-seizure uses for the newer anti-epileptic drugs. AB - Anti-epileptic drugs are being used to treat a variety of conditions. This column provides an overview of the potential uses of the new AEDs. More randomized, clinical trials are needed before considering the new AEDs as first-line therapy for non-seizure disorders, but these agents appear to be emerging as potential options for a variety of conditions. PMID- 11016275 TI - Recognition, assessment and management of neuroleptic malignant syndrome. AB - Neuroleptic malignant syndrome (NMS) is a potentially fatal idiosyncratic complication of neuroleptic treatment. It was first described in 1968 by Delay and Deniker, and is characterized by hyperthermia, extrapyramidal signs, autonomic dysfunction and altered mentation. They reported that 0.5% to 1.0% of the patients on neuroleptic therapy developed NMS. Several retrospective studies have reported the frequency of NMS between 0.02% to 2.44%. These variations in the incidence of NMS may be due to difference in patient population, awareness and experience of clinicians, prescribing habits, different diagnostic criteria, or methodological differences in survey methods. PMID- 11016276 TI - A brief history of the South Dakota Society of Obstetrics and Gynecology. PMID- 11016277 TI - [Can sudden death in epilepsy be prevented?]. PMID- 11016278 TI - [Diagnostic and therapeutic dilemmas in type 2 diabetes]. PMID- 11016279 TI - [Investigation and treatment of idiopathic scoliosis in adolescence]. PMID- 11016280 TI - [Mortality in epilepsy. A review]. AB - The overall risk of premature death in patients with epilepsy is two to three times that of the general population. The mortality risk is highest in patients with symptomatic epilepsy, but the mortality rate is also increased in patients with idiopathic epilepsy, indicating that epilepsy itself carries an increased risk of premature death. Cerebrovascular disease and primary brain tumours are common causes of death in patients with symptomatic epilepsy, while sudden unexplained death (SUDEP), accidents, suicide and status epilepticus are important causes of epilepsy related deaths. Prevention of accidents and suicide in patients with epilepsy and further knowledge on SUDEP is essential in order to reduce the mortality rate of epilepsy. PMID- 11016281 TI - [Intensified treatment of type 2 diabetes mellitus. Is polypharmacy necessary and justified?]. AB - Newly published randomised controlled trials with pharmacological intervention against hyperglycaemia, hypertension and dyslipidemia have challenged the traditional empiric treatment of type 2-diabetes. This review focuses on the results of these trials as well as randomised trials with pharmacological therapy of microalbuminuria, primary prevention with acetylsalicylic acid and angiotensin converting enzyme (ACE) inhibitors. The overall results from these trials are clinically relevant reductions in the risk of late diabetic complications. Taken together, the new clinical knowledge does not mean that all patients with type 2 diabetes besides relevant changes in lifestyle will benefit from a comprehensive polypharmacy. It means, however, that based upon the individual risk profile the medical professionals have to motivate the patient for an evidence based "therapeutic package" which is likely to improve the longterm outcome. PMID- 11016282 TI - [Legionella pneumophila in pool water]. AB - We investigated 87 samples of pool water for the presence of Legionella. The samples were from cold water pools (< 28 degrees C) and from hot water pools (> 32 degrees C). Sampling was furthermore done on normal water samples, on water from the bottom of pools and of water from departure from the activated carbon filters. Legionella was not detected in any of the samples from cold water pools, whereas in the hot water pools 10% of the pool water samples and 80% of the water from the filters were infected with Legionella pneumophila. The highest concentrations of Legionella were found in the filter samples, whereas the concentrations in the pool water (10-100 cfu/liter) were not alarming. This investigation demonstrates the potential risk of presence of Legionella in hot water pools with activated carbon filters being the site with best growth potential. It should be stressed that a high level of disinfection (at least 1.0 mg free chlorine/liter and pH 7.2) is essential for the prevention of Legionella in pool water at higher temperatures. PMID- 11016283 TI - [Prevalence of idiopathic scoliosis in the municipality of Hillerod]. AB - Among Scandinavian paediatric spinal surgeons there has been a debate whether the prevalence of idiopathic adolescent scoliosis (AIS) has declined. We examined all children in the town of Hillerod, Denmark attending third and fifth grade (age 10 and 12) with forward-bending-test using a scoliometer. All children with more than seven degrees of trunk inclination were referred to a PA radiogram of the spine. We found a 0.4 percent prevalence of AIS with Cobbangles greater than 19 degrees. This is similar to earlier findings, suggesting that the declining referral rate is due to late detection of idiopathic adolescent scoliosis. PMID- 11016284 TI - [Hereditary hemorrhagic telangiectasia. A population-based study on prevalence and mortality among Danish HHT patients]. AB - Hereditary haemorrhagic telangiectasia (HHT) is a dominantly inherited disease characterized by telangiectatic lesions. The disease manifestations are variable in severity. Early death due to complications has been described. We report an investigation of the prevalence and mortality of HHT in a Danish population, based on two cross-sectional surveys in combination with a long-term follow-up study. The prevalence of HHT in the county of Funen was 13.8 per 100,000 January 1. 1974 and 15.6 per 100,000 January 1. 1995. In the HHT group as a whole we found a slightly increased mortality, however among the HHT patients younger than 60 years at inclusion the mortality of HHT patients was twice the expected. The excess mortality was fully explained by severe HHT symptoms contributing to death. Future research should aim at identification of HHT patients at particular risk of developing severe complications. PMID- 11016285 TI - ["Danger spots" for bicyclists based on data from accident registration in emergency departments]. AB - In Denmark political decisions to reduce traffic morbidity are often based on data collected from the police. It is often claimed that hospital-based traffic accident registration cannot provide detailed information regarding the exact localization of the accident. On the other hand, several investigations have shown a coverage of bicyclist accidents of only 8-19% in police records when comparing with hospital-based records. In the period from 1st of February 1995 until 31st of January 1997 all 971 patients injured in bicycle accidents and treated at the Emergency Department at Viborg Hospital were given a questionnaire regarding details about the accident and the exact localization of the accident. Eighty-five percent answered the questionnaire. Eight percent of the accidents were also found in the police register. From the material it could be concluded that six roads in the Municipality of Viborg were the scene of the accident in more than 10 cases each. A further analysis of these accidents pointed out two exact localizations on one single road as especially hazardous for bicyclists. It is concluded, that police-based registration of bicycle accidents are insufficient and more resources should be used on hospital-based accident registration, since detailed information about the site of the accidents also can be provided there. PMID- 11016286 TI - [Treatment of tinnitus with low-intensity laser]. AB - This study evaluated the effect of low-power laser in the treatment of tinnitus in a randomized, prospective, double-blind, placebo-controlled trial. The active laser applied 50 mW (cw, 830 nm) over a period of 10 minutes per session. Forty nine patients were included. The main outcome was measured using psychoacoustical match of tinnitus loudness, Visual Analog Scale (VAS) ratings of subjective loudness, annoyance and attention involved, scores on tinnitus-specific questionnaires, and a number of psychosocial questionnaires. Only few subjects (18%) experienced subjective improvement. There were no statistically significant differences between the effects of the active laser and placebo treatments. CONCLUSION: Low-power laser treatment is not indicated in the treatment of tinnitus. Reports of significant benefits of this treatment in previous studies may be explained by the placebo effect. PMID- 11016287 TI - [Characteristics and treatment of acute pulmonary embolism. A study of 183 consecutive patients treated during the period 1995 to 1998]. AB - The aim of the present study was to evaluate the clinical characteristics and mortality in patients with pulmonary embolism during 1995-1998. The diagnosis of pulmonary embolism in 183 patients was confirmed based on the clinical and lung scan findings. Compared to previous studies fewer cases with pulmonary embolism after surgery, immobilisation and history of trauma to lower extremity were noted. Deep vein thrombosis and electrocardiographic signs of acute right ventricular strains were found frequently, and should support the suspicion of pulmonary embolism. A normal plasma fibrin D-dimer was noted in several patients. When using the D-dimer for the diagnosis of pulmonary embolism the result given depends on the assay method used, the assay specific discriminatory level and duration of symptoms. The total one year mortality was 16% whereas the mortality due to pulmonary embolism was 6%. PMID- 11016289 TI - [Pseudotumor in the parotid region]. AB - Tophaceous pseudogout is a rare form of calcium pyrophosphate dihydrate (CPPD) crystal deposition disease. Half of the 31 reported cases of tophaceous pseudogout are in the head and neck region. This patient presented with a parotid tumour, that was initially suspected to be malignant based on radiology and cytology. Operation disclosed a tumour progressing to the base of the skull, histological examination showed inflammatory cells, macrophages, metaplastic chondroid cells and giant cells of foreign-body-type. X-ray diffraction revealed two crystal forms of CPPD. PMID- 11016288 TI - [Celiac disease with subepithelial deposition of collagen]. AB - In 1970 a case of malabsorption with flat small intestinal mucosa with subepithelial collagen deposition was described. There was no response to a gluten-free diet, and the condition was termed collagenous sprue. We report a case of coeliac disease with subepithelial deposition of collagen in duodenal biopsy, which responded to a gluten-free diet. PMID- 11016290 TI - [Picture of the month]. PMID- 11016291 TI - [The iron gate is open--by a transparent zebrafish]. PMID- 11016292 TI - [No to patents for genetic diagnostics]. PMID- 11016293 TI - [The average longevity of Danes: significance of health care]. PMID- 11016294 TI - [Consciousness during anesthesia]. PMID- 11016295 TI - Physician leadership in cardiac outcomes reporting. PMID- 11016296 TI - Washington State's model of physician leadership in cardiac outcomes reporting. AB - BACKGROUND: In 1993, the cardiac surgery community in Washington State opposed an effort by the state Health Care Authority (HCA) to identify "centers of excellence" for selective contracting of coronary artery bypass grafting (CABG) procedures, and proposed an alternate model that would create a statewide cardiac outcomes registry under physician governance to be used by all institutions for internal quality improvement activities. METHODS: A prospective pilot data collection effort, which examined preoperative and postoperative patient-reported health status, served as the basis for evaluating the capacity of a physician-led organization to develop a collaborative atmosphere and facilitate universal hospital participation. RESULTS: A surgical steering group met on a regular basis and reached consensus on governance issues, protocols for standardized data collection, and policies regarding data dissemination. All 14 centers that performed bypass surgery in the state participated. Patients who were surveyed reported statistically significant improvements in physical, emotional, and anginal-specific health status after bypass surgery. Baseline patient characteristics and longitudinal outcomes were compared across institutions. CONCLUSIONS: Based on the feasibility of this collaborative outcomes reporting program, the HCA revised its policy regarding selective contracting and has helped to support an ongoing physician-led and -governed cardiac outcomes reporting system that is particularly notable for the subsequent integration of both CABG surgery and catheterization-based procedures into one standardized registry. PMID- 11016297 TI - Identification of risk factors for increased cost, charges, and length of stay for cardiac patients. AB - BACKGROUND: In this study we explored different risk model options to provide clinicians with predictions for resource utilization. The hypotheses were that predictors of mortality are not predictive of resource consumption, and that there is a correlation between cost estimates derived using a cost-to-charge ratio or a product-line costing approach. METHODS: From March 1992 to June 1995, 2,481 University of Colorado Hospital patients admitted for ischemic heart disease were classified by diagnosis-related group code as having undergone or experienced coronary bypass procedures (CBP), percutaneous cardiovascular procedures (PCVP), acute myocardial infarction (AMI), and other cardiac-related discharges (Other). For each diagnosis-related group, Cox proportional hazards models were developed to determine predictors of cost, charges, and length of stay. RESULTS: The diagnosis groups differed in the clinical factors that predicted resource use. As the two costing methods were highly correlated, either approach may be used to assess relative resource consumption provided costs are reconciled to audited financial statements. CONCLUSIONS: To develop valid prediction models for costs of care, the clinical risk factors that are traditionally used to predict risk-adjusted mortality may need to be expanded. PMID- 11016298 TI - Are bilateral superior vena cavae a risk factor for single ventricle palliation? AB - BACKGROUND: Performing superior vena cava-to-pulmonary artery anastomosis, in the presence of bilateral superior vena cavae, can be technically challenging. Our clinical observation has been that bilateral superior vena cavae are a risk factor for poor outcome in children needing single ventricle palliation. METHODS: Detailed operative, angiographic, and follow-up data were analyzed in 39 children undergoing bilateral cavopulmonary anastomosis (b-CPA). Overall outcome was compared to 274 children having a unilateral cavopulmonary anastomoses (u-CPA). RESULTS: Nine patients (23%) with bilateral superior vena cavae were found to have thrombus in the cavopulmonary circulation after the b-CPA. Postoperative mean arterial oxygen saturation was significantly lower in those who had thrombus [69%+/-10% versus 82%+/-7%, (p < 0.01)]. Thrombus formation was associated with mortality. The indexed superior vena cavae size was not a risk factor for thrombosis. In follow-up studies the connecting pulmonary artery segment between the two cavopulmonary anastomosis was smaller than the pulmonary arteries adjacent to the hilum. Survivors of a b-CPA were less frequently converted to a Fontan circulation at 5 years of follow up (Kaplan-Meier 5-year estimates, 39% for b-CPA versus 74% for u-CPA [p = 0.02]). CONCLUSIONS: Bilateral superior vena cava-to-pulmonary artery anastomosis is associated with an increased risk of thrombus formation and unfavorable growth in the central pulmonary arteries. Modifications of surgical technique may alter flow patterns, thereby optimizing growth and diminishing the risk of thrombus formation. Anticoagulation therapy may be an important adjunct in children undergoing a b-CPA. PMID- 11016299 TI - Comparison of porcine xenografts and homografts for pulmonary valve replacement in children. AB - BACKGROUND: Due to the limited availability of homografts, different alternatives are used for replacement of the pulmonary valve. This study investigates the value of porcine stentless pulmonary xenografts in pediatric cardiac patients. METHODS: Twenty-three pediatric xenograft (size 10 to 21 mm) recipients were compared with 23 homograft (size 9 to 21 mm) recipients. RESULTS: Hospital mortality was 2 of 23 patients in the xenograft group and 3 of 23 in the homograft group (NS). Six out of 20 xenografts and 1 of 19 homografts were stenotic after 1 year (p = 0.011). Xenograft stenoses were mainly located at the distal anastomosis, while the leaflets were preserved. Homografts showed valvular stenoses and wall calcification. The 1 year freedom from reoperation was 77% in the xenograft and 93% in homograft recipients (NS), and from transcatheter intervention 84% and 100% (p = 0.004), respectively. Transcatheter intervention in 7 xenograft patients and 1 homograft recipient improved stenosis gradients from 65 to 40 mm Hg (mean) in 6 out of 8 patients. Explanted xenografts showed a loss of elastic membranes and proliferating connective tissue scars coated with activated endothelium. CONCLUSIONS: Xenografts demonstrated a higher incidence of supravalvular obstructions, which were possibly due to unfavorable hemodynamics at the distal anastomosis. Histological findings additionally indicated a pronounced immunological response. Interventional angioplasty lowered the rate of reoperation. Thus, the use of xenografts in children can be accepted as a second choice when a homograft is unavailable. PMID- 11016300 TI - Right ventricular outflow tract after non-conduit repair of tetralogy of Fallot with coronary anomaly. AB - BACKGROUND: A total of 25 patients with tetralogy of Fallot and an important coronary artery crossing the right ventricular outflow tract underwent complete repair without use of an extracardiac conduit between January 1990 and December 1994. Repair was exclusively done by the transatrial or transatrial transpulmonary approach. Age of these patients ranged from 1 to 12 years (mean 3.6 years). Three of the patients had already received a systemic to pulmonary artery shunt. METHODS: All patients reporting for follow-up (n = 18) were subjected to transthoracic echocardiography and, if required, cardiac catheterization and angiography. Right ventricle to pulmonary artery gradients were noted preoperatively, at discharge following repair and at follow-up study. RESULTS: Mean follow-up was 40.6 months (24 to 62 months). Mean early postoperative gradient was 23.5+/-13.4 mm Hg and 4 patients had significant (> 30 mm Hg) gradients. Mean late postoperative gradient was 20.6+/-12.4 mmHg and 2 patients had gradients greater than 30 mmHg. All the patients were in New York Heart Association functional class I at the time of last follow-up. CONCLUSIONS: Acceptable gradients across the right ventricular outflow tract are achievable following repair of tetralogy of Fallot in the presence of anomalous coronary artery across the right ventricular outflow tract using the transatrial or transatrial-transpulmonary approach. Most gradients were found not to vary significantly on subsequent follow-up. PMID- 11016301 TI - The long-term outcome of a surgical repair of sinus of valsalva aneurysm. AB - BACKGROUND: In order to clarify the long-term outcome after surgical repair of a sinus of Valsalva aneurysm, we retrospectively assessed the operative results for patients treated in our institute. METHODS: The subjects were 27 patients who had undergone an operation between 1958 and 1996. For associated aortic regurgitation (AR) aortic valve repair was performed in 13 patients, 12 of whom had a ventricular septal defect (VSD); and an aortic valve replacement was performed in 3 patients, 1 of whom had a VSD. RESULTS: Five of the 13 patients who had aortic valve repair needed aortic valve replacement because AR developed after a period of between 7 and 13 years; those cases were complicated by VSD. Another 2 patients with mild AR also complicated by VSD are currently under observation. CONCLUSIONS: Although the postoperative outcome of the aortic valve repairs was good, cases that were complicated by VSD plus associated AR tended to develop AR later after surgery. Therefore, careful observation of the postoperative course is necessary. PMID- 11016302 TI - Video-assisted cardioscopy for intraventricular repair in congenital heart disease. AB - BACKGROUND: Video-assisted thoracoscopic surgical techniques have been widely adopted as a means to reduce surgical trauma. By adapting pediatric thoracoscopic instrumentation, we have developed a technique for video-assisted cardioscopy (VAC). We report our experience and describe the technical feasibility of VAC. METHODS: Since June 1995, 409 consecutive patients underwent 431 intracardiac procedures (ventricular septal defect, 150; tetralogy of Fallot or double outlet right ventricle, 101; atrioventricular canal, 52; subaortic stenosis, 43; valve repair, 50; Rastelli procedure, 12; Konno or Ross Konno operation, 11; and miscellaneous, 12) using VAC at Miami Children's Hospital. Using a prospective database, we tracked outcomes and operative events to delineate the usefulness and efficacy of this technique. RESULTS: VAC provided clear and precise imaging of small or remote intracardiac structures during repair of congenital heart defects without technical complications. Procedure times and aortic cross-clamp times using VAC were not prolonged. Intraoperative images were collected for every operation, documenting each patient's cardiac anatomy before and after repair. Surgery through small incisions was facilitated. Operative mortality was 1.2% (5 of 409), and no patient required reoperation before discharge. At a mean follow-up interval of 22 months, the incidence of reoperation for residual or recurrent lesions was 1.2% (5 of 404). CONCLUSIONS: Our experience demonstrates the technical feasibility and clinical utility of routine endoscopic imaging during open heart surgery for congenital heart repair. PMID- 11016303 TI - Repair of cardiac defects through a shorter right lateral thoracotomy in children. AB - BACKGROUND: Median sternotomy is a conventional approach for correction of cardiac defects for many years; however, the cosmetic result is poor. Therefore, right lateral thoracotomy was tested as an alternative procedure with a better cosmetic outcome. METHODS: Between October 1994 and February 1999, 683 patients underwent correction of congenital cardiac malformations during a cardiopulmonary bypass through right lateral thoracotomy involving a shorter incision through the third or the fourth intercostal space. All of the patients were children. The average age was 3.26+/-1.67 years (range, 4 months to 7 years). The average weight was 13.59+/-4.37 kg (5 to 40). The patients had various cardiac defects and associated anomalies. RESULTS: Only 2 patients died after operation, 1 from low cardiac output and the other from severe pulmonary infection. The hospital morbidity was lower. The mean cardiopulmonary bypass time was 58.67+/-35.11 minutes (range, 16 to 430 minutes), the mean aortic cross-clamping time was 35.03+/-24.84 minutes (range, 3 to 205 minutes). The postoperative average mechanical ventilation time was 19.23+/-39.11 hours (range, 2 to 391 hours), and the mean postoperative stay was 8.55+/-12.54 days (range, 5 to 293 days). CONCLUSIONS: The right lateral thoracotomy incision is a safe and effective alternative to a median sternotomy for correction of cardiac defects. Advantages of this approach compared with median sternotomy are less injury, maintenance of the continuity and the integrity of the bony thorax, and prevention of the development of "pigeon-chesting." The scar is less visible, hence, the cosmetic result can meet patient expectations. This procedure is consistent with the idea of minimal invasive surgery. PMID- 11016304 TI - Rapid cardiopulmonary support for children with complex congenital heart disease. AB - BACKGROUND: Extracorporeal membrane oxygenation has limitations in children with congenital heart disease (prolonged setup times, increased postoperative blood loss, and difficulty during transport). We developed a miniaturized cardiopulmonary support circuit to address these limitations. PATIENTS AND METHODS: The cardiopulmonary support system includes a preassembled, completely heparin-coated circuit, a BP-50 Bio-Medicus centrifugal pump, a Minimax plus membrane oxygenator, a Bio-Medicus flow probe, and a Bio-trend hematocrit/oxygen saturation monitor. Short tubing length permits a 250-mL bloodless prime in less than 5 minutes. From 1995 to 1997, 23 children with congenital heart disease were supported with this technique. RESULTS: Overall survival to discharge was 48% (11 of 23 patients). Survival to discharge was 80% (4 of 5) in the preoperative support group, 20% (1 of 5) in the postoperative failure to wean from cardiopulmonary bypass group, 44% (4 of 9) in the group placed on support postoperatively after transfer to the intensive care unit, and 50% (2 of 4 patients) in the nonoperative group. Neonatal cardiopulmonary support survival to discharge was 46% (6 of 13 patients). CONCLUSIONS: This pediatric cardiopulmonary support system is safe and effective. Advantages over conventional extracorporeal membrane oxygenation include rapid setup time, decreased postoperative blood loss, and simplified transport. PMID- 11016305 TI - Hyperoxia for management of acid-base status during deep hypothermia with circulatory arrest. AB - BACKGROUND: Which blood gas strategy to use during deep hypothermic circulatory arrest has not been resolved because of conflicting data regarding the advantage of pH-stat versus alpha-stat. Oxygen pressure field theory suggests that hyperoxia just before deep hypothermic circulatory arrest takes advantage of increased oxygen solubility and reduced oxygen consumption to load tissues with excess oxygen. The objective of this study was to determine whether prevention of tissue hypoxia with this strategy could attenuate ischemic and reperfusion injury. METHODS: Infants who had deep hypothermic circulatory arrest (n = 37) were compared retrospectively. Treatments were alpha-stat and normoxia (group I), alpha-stat and hyperoxia (group II), pH-stat and normoxia (group III), and pH stat and hyperoxia (group IV). RESULTS: Both hyperoxia groups had less acidosis after deep hypothermic circulatory arrest than normoxia groups. Group IV had less acid generation during circulatory arrest and less base excess after arrest than groups I, II, or III (p < 0.05). Group IV produced only 25% as much acid during deep hypothermic circulatory arrest as the next closest group (group II). CONCLUSIONS: Hyperoxia before deep hypothermic circulatory arrest with alpha-stat or pH-stat strategy demonstrated advantages over normoxia. Furthermore, pH-stat strategy using hyperoxia provided superior venous blood gas values over any of the other groups after circulatory arrest. PMID- 11016306 TI - The role of cardioplegia induction temperature and amino acid enrichment in neonatal myocardial protection. AB - BACKGROUND: Warm cardioplegic induction improves the ischemically "stressed" adult heart. However, it is rarely used in infants, despite the fact that many newborn hearts are stressed by other factors such as hypoxia. The need for amino acids as well as their mechanism of action has also not been studied. METHODS: We first assessed the role of cardioplegic induction temperature in 10 nonhypoxic neonatal piglets undergoing 70 minutes of multidose blood cardioplegic arrest. Five piglets (group 1) received a cold (4 degrees C) induction, and 5 (group 2) a warm (37 degrees C) induction. Twenty-six other piglets underwent ventilator hypoxia (fraction of inspired oxygen, 8% to 10%) for 60 minutes before cardiopulmonary bypass (stress). Six piglets (group 3) then underwent 70 minutes of cardiopulmonary bypass without ischemia (hypoxia controls), and 20 underwent 70 minutes of cardioplegic arrest. Five of these (group 4) received cold cardioplegic induction, and 15 received warm induction; in 5 of these (group 5), the warm cardioplegic solution contained amino acids, in 5 others (group 6), it was unsupplemented, and in the remaining 5 (group 7), nitroglycerin was added to determine the role of vasodilation. Myocardial function was assessed by pressure volume loops (expressed as a percent of control), and coronary vascular resistance was measured with cardioplegic infusions. RESULTS: In nonhypoxic (normal) piglets, cold (group 1) and warm (group 2) induction completely preserved systolic function (end-systolic elastance, 100% versus 104%) and preload recruitable stroke work (100% versus 102%), with minimal increase in diastolic compliance (162% versus 156%). Hypoxia-reoxygenation alone (group 3) depressed systolic function (end-systolic elastance, 51%+/-2%) and preload recruitable stroke work (54%+/-3%), and raised diastolic stiffness (260%+/-15%). The detrimental effects of reoxygenation persisted (unchanged from reoxygenation alone) with cold induction (group 4) or warm induction without amino acids (groups 6 and 7). In contrast, warm induction with amino acids (group 5) restored systolic function (end-systolic elastance, 105%+/-3%; p < 0.001 versus groups 3, 4, 6, and 7) and preload recruitable stroke work (103%+/-2%; p < 0.001 versus groups 3, 4, 6, and 7), and decreased diastolic stiffness (154%+/-7%; p < 0.001 versus groups 3, 4, 6, and 7). However, there was no difference in myocardial oxygen consumption in hypoxic hearts receiving a warm induction (6.9 versus 6.5 versus 7.3 mL/g per 5 minutes) (groups 5, 6, 7), and coronary vascular resistance was lowest with nitroglycerin (group 7). CONCLUSIONS: Cardioplegic induction can be given either warm or cold in nonhypoxic neonatal hearts. In contrast, only warm induction with amino acids repairs the hypoxic injury, but the primary mechanism of action is not related to increased metabolic activity or vasodilation. PMID- 11016307 TI - Reducing postischemic reperfusion damage in neonates using a terminal warm substrate-enriched blood cardioplegic reperfusate. AB - BACKGROUND: In adult cardiac operations, a warm cardioplegic reperfusate ("hot shot") before removing the aortic cross-clamp improves postbypass myocardial function and metabolic recovery. This modality, however, is rarely used in infants, despite the fact that postbypass cardiac dysfunction remains problematic, especially in cyanotic ("stressed") patients. METHODS: To produce stress, 15 neonatal piglets underwent 60 minutes of ventilator hypoxia (fraction of inspired oxygen, 8% to 10%). All piglets then received similar protection with multidose cold blood cardioplegic solution during 70 minutes of arrest and were separated into three groups to examine the role of a warm reperfusate as well as possible augmentation by aspartate and glutamate enrichment. In 5 piglets (group 1), the cross-clamp was simply removed; in 5 (group 2), an unsupplemented warm blood cardioplegic reperfusate was given; and in 5 (group 3), the warm reperfusate was enriched with aspartate and glutamate. Myocardial function was assessed using pressure-volume loops and expressed as a percentage of control. RESULTS: Compared with hearts receiving reperfusion with unmodified blood (group 1), a warm unsupplemented cardioplegic reperfusate (group 2) slightly improved systolic contractility (end-systolic elastance, 41% versus 50%; p < 0.05 versus group 1) and preload recruitable stroke work (41% versus 52%; p < 0.05 versus group 1), reduced diastolic stiffness (263% versus 245%; p < 0.05 versus group 1), and increased adenosine triphosphate (10.7 versus 11.9 microg/g tissue, p < 0.05 versus group 1). However, if aspartate and glutamate was included in the warm reperfusate (group 3), there was complete recovery of systolic function (end systolic elastance, 105%+/-3%; p < 0.001 versus all groups) and preload recruitable stroke work (103%+/-2%; p < 0.001 versus all groups), a minimal rise in diastolic stiffness (154%+/-7%; p < 0.001 versus all groups), and preservation of adenosine triphosphate (15.5+/-0.5 microg/g; p < 0.001 versus all groups). CONCLUSIONS: A warm cardioplegic reperfusate helps reduce the reperfusion injury, resulting in improved myocardial function and metabolic recovery in hypoxic (stressed) neonatal hearts, and this effect is maximized if the reperfusate is enriched with aspartate and glutamate, which completely preserves myocardial function. PMID- 11016308 TI - Papillary muscle repair surgery in ischemic mitral valve patients. AB - BACKGROUND: Ischemic mitral regurgitation (MR), when ischemia/infarction has resulted in fibrotic degeneration and elongation of papillary muscles, carries a high risk for the patient and a technical challenge for the surgeon. We have developed a papillary-shortening plasty for this specific pathology. METHODS: Papillary muscle repair was performed in 88 patients (7.2%) where degenerated and fibrotic elongated papillary muscles were found, which resulted in a prolapse of one or more parts of the mitral valve leaflets (MR III-IV). All patients had a papillary muscle-shortening plasty using a pericardium pledged-reinforced polytetrafluoroethylene suture and a ring annuloplasty. Because the cause of regurgitation in this specific group of patients was ischemic, concomitant coronary bypass grafting was required in all patients, with 2.2 grafts/patient. RESULTS: There were five hospital deaths (5.7%). Postoperative mitral valve function was satisfactory in all patients: no residual mitral regurgitation (MR 0) was found in 80 patients (90.9%), mild regurgitation (MR I) in 5 patients (5.7%), and moderate regurgitation (MR I-II) was observed in 3 patients (3.4%). Within a short mean follow-up period of 18.6 months (3 to 40 months), there was one late death (1.2%). The actuarial freedom from reoperation and thromboembolic complications was 100%, but there were two anticoagulation-induced gastric bleeding complications (2.3%). All patients were in New York Heart Association functional class I or II at the time of follow-up. CONCLUSIONS: Our data show that careful assessment of papillary muscle pathology is mandatory, and that a papillary muscle-shortening plasty is a simple but valuable surgical tool to repair the mitral valve in this specific group of high-risk patients with ischemic mitral regurgitation. PMID- 11016309 TI - Influence of concomitant CABG and urgent/emergent status on mitral valve replacement surgery. AB - BACKGROUND: Outcomes and resource utilization of patients undergoing mitral valve replacement (MVR) with or without concomitant coronary artery bypass grafting (CABG) were reviewed. METHODS: Data for 1,844 patients undergoing isolated primary MVR at Emory University Hospitals between 1980 and 1997 were recorded prospectively in a computerized database. RESULTS: The four groups included patients undergoing elective MVR with (n = 360) or without CABG (n = 1332) and urgent/emergent MVR with (n = 66) or without CABG (n = 86). Length of stay was significantly higher in patients undergoing elective MVR with CABG (15 days) than in those without CABG (11 days) but was not significantly different in patients undergoing urgent/emergent MVR with CABG (17 days) than in those without CABG (19 days). In-hospital mortality was significantly higher for patients undergoing elective (14%) or urgent/emergent (41%) MVR with CABG than in those undergoing MVR without CABG (elective:6%; urgent/emergent:20%). The 19-year survival rate was 32% for patients undergoing elective MVR with CABG compared with 51% for those without CABG and 28% for patients undergoing urgent/emergent MVR with CABG compared with 46% for those without CABG. Multivariate correlates of long-term mortality included older age, concomitant CABG, and urgent/emergent status. Hospital costs were significantly higher for patients undergoing elective MVR with ($33,216) than for those without ($23,890) CABG. No significant difference in cost were noted between patients undergoing urgent/emergent MVR with ($40,535) and without ($31,981) CABG. CONCLUSIONS: The addition of CABG or urgent/emergent status to patients undergoing MVR significantly increases morbidity, mortality, and costs. Careful scrutiny of the benefits versus resource utilization is required for patients undergoing high risk MVR. PMID- 11016310 TI - Survival advantage of stentless aortic bioprostheses. AB - BACKGROUND: Bioprostheses (BPs) are used to avoid anticoagulation after aortic valve replacement (AVR) in patients over 65 years of age. Stentless BPs offer established hemodynamic benefits. We sought to determine whether these advantages translate into improved survival. METHODS: Between 1993 and 1997, follow-up data (for Food and Drug Administration submission) were collected prospectively for 160 consecutive, unselected hospital survivors who received the Freestyle valve (FS). Equivalent data were collected for 247 Carpentier-Edwards (CE) porcine xenograft patients. Detailed comparative statistical analysis was used to compare events and survival between the groups. Follow-up was 100% complete for the FS (5.2 years maximum; mean 3.2+/-1.0 years) group and 98% (7.2 years maximum; mean 3.8+/-2.0 years) for CE. RESULTS: The groups were well matched in age (FS, 73+/-6 years; CE, 74+/-6 years), gender (FS, 58% male; CE, 62% male), ventricular function, and number of patients requiring coronary grafts (FS, 41%; CE, 37%). Actuarial survival at 5 years was 84% for FS versus 69% for CE (p = 0.023 Kaplan Meier, p = 0.009 Cox). Annual mortality rates were 3.6% for FS versus 7.1% for CE (p = 0.001). Thromboembolic rate was 0.8% per year for FS and 2.4% for CE (p = 0.024) without a difference in cardiac rhythm. Incidence of nonstructural dysfunction (paravalvular leak) was 0.2% for FS versus 1.3% for CE (p = 0.020). CONCLUSIONS: By 5 years, the stentless valve patients had improved survival and reduced adverse events. Though differences in durability are yet to be proved, our findings support the use of stentless bioprostheses in this age group. PMID- 11016311 TI - Viability of cryopreserved semilunar valves: an evaluation of cytosolic and mitochondrial activities. AB - BACKGROUND: Despite long-standing, widespread use of cryopreserved allografts, the basic cellular biology of these tissues is still yet unknown. The present investigation was undertaken to study cryopreserved heart valves from the standpoint of cytosolic esterase and mitochondrial dehydrogenase activities. METHODS: Cryopreserved porcine aortic cusps were observed in an unfixed fresh condition with a confocal laser scanning microscope using fluorescent dye. Porcine cusps and cultured human umbilical vein endothelial cells were divided into three groups, including fresh, cold-preserved, and cryopreserved specimens, and cytosolic esterase activity and mitochondrial dehydrogenase activity were analyzed in each. RESULTS: Confocal laser scanning microscope findings disclosed a widely distributed fluorescence in the cusp. Cytosolic esterase activity within human umbilical vein endothelial cells (28%+/-9.0%) after cryopreservation was significantly less than that it was in the cusps (72%+/-21%). Mitochondrial dehydrogenase activity of cryopreserved human umbilical vein endothelial cells and that of cusps fell to 44%+/-6.1% and 64%+/-17% respectively; the difference between the two values was not significant. CONCLUSIONS: Cryopreservation appeared to produce serious damage to cytosolic and mitochondrial functions of endothelial cells. The cytosolic function of cusps, mainly consisting of fibroblasts, was comparatively preserved after cryopreservation, but mitochondrial function of the cusps was more diminished. PMID- 11016312 TI - Long-term follow-up after Carpentier-Edwards ring annuloplasty for tricuspid regurgitation. AB - BACKGROUND: Use of flexible rings for tricuspid ring annuloplasty is becoming popular. This study was undertaken to evaluate Carpentier-Edwards (C-E) rigid ring annuloplasty for tricuspid regurgitation (TR), secondary to mitral valve disease and clinical outcome on a long-term basis. METHODS: From December 1985 to March 1996, 45 patients with secondary TR underwent C-E ring annuloplasty. Thirty nine patients (95.1%) were in New York Heart Association (NYHA) functional class III or IV. The mean follow-up was 96.7+/-48.5 months or 362.6 patient-years. RESULTS: There were three in-hospital and nine late deaths that were not related to tricuspid annuloplasty. Actuarial survival at 10 years was 68.3%. Echocardiographic studies showed that TR was well controlled within grade 2+ in all survivors. Residual pulmonary hypertension (PH) was recognized in 9 of 21 patients (42.9%) with preoperative PH, however, no TR was seen in 6 patients. A TR grade of 2+ was observed in 3 patients. Thirty of the total survivors (96.8%) were in NYHA class I and II, but 1 patient was in NYHA class III. The actuarial rate of freedom from tricuspid valve reoperation after 10 years was 97.5%. CONCLUSIONS: C-E ring annuloplasty is acceptable for repair of secondary TR and improvement in clinical status on a long-term basis. PMID- 11016313 TI - The influence of gender on the outcome of coronary artery bypass surgery. AB - BACKGROUND: To assess the impact of gender as an independent risk factor for early and late morbidity and mortality following coronary artery bypass surgery. METHODS: Perioperative and long-term data on all 4,823 patients undergoing isolated coronary bypass operations from November 1989 to July 1998 were analyzed. Of these patients, 932 (19.3%) were females. RESULTS: During the years 1989 to 1998 there was a progressive increase in the percentage of women undergoing coronary artery bypass surgery. The following preoperative risk factors were more prevalent among women than men: age above 70, angina class 3 or 4, urgent operation, preoperative intraaortic balloon pump usage, congestive heart failure, previous percutaneous transluminal coronary angioplasty, diabetes, hypertension, and peripheral vascular disease (all p < 0.05). Men were more likely to have an ejection fraction less than 35%, three-vessel disease, repeat operations, and a recent history of smoking. Women had a statistically significant smaller mean body surface area than men (1.72+/-0.18 versus 1.96%+/ 0.26% m2). On average, women had fewer bypass grafts constructed than men (2.9%+/ 0.8% versus 3.2%+/-0.9%) and were less likely to have internal mammary artery grafting (76.2% versus 86.1%), multiple arterial conduits (10.1% versus 19.8%), or coronary endarterectomy performed (4.9% versus 8.6%). The early mortality rate in women was 2.7% versus 1.8% in men (p = 0.09). Women were more prone to perioperative myocardial infarction (4.5% versus 3.1% p < 0.05). After adjustment for other risk variables, female gender was not an independent predictor of early mortality but was a weak independent predictor for the prespecified composite endpoint of death, perioperative myocardial infarction, intraaortic balloon counterpulsation pump insertion, or stroke (8.55 versus 5.9%; odds ratio, 1.30; 95% confidence interval, 0.99 to 1.68; p = 0.05) Recurrent angina class 3 or 4 was more frequent in female patients (15.2%+/-4.0% versus 8.5%+/-2.0% at 60 months, p = 0.001) but not repeat revascularization procedures (percutaneous transluminal coronary angioplasty, redo) (0.6%+/-0.3% versus 4.1%+/-0.8% at 60 months). Actuarial survival at 60 months was greater in women then men (93.1%+/ 1.7% versus 90.0%+/-1.0%), and after adjustment for other risk variables, female gender was protective for late survival (risk ratio, 0.40; 95% confidence interval, 0.16-0.74; p < 0.005). CONCLUSIONS: Perioperative complications were increased and recurrent angina more frequent in women. Despite this, late survival was increased in women compared with men after adjustment for other risk variables PMID- 11016314 TI - Reperfusion injury associated with one-fourth of deaths after coronary artery bypass grafting. AB - BACKGROUND: This study of reperfusion injury after coronary artery bypass grafting focuses on its contribution to fatal outcome, on its connection with myocardial infarction (MI) and on risk factors. METHODS: A consecutive series of 190 patients (mean age 61.7+/-8.9 years) dying within 30 days following coronary artery bypass grafting was autopsied with concomitant postmortem angiography during 1980 to 1993. RESULTS: Reperfusion injury was revealed in 49 (25.8%) patients, with concomitant MI in almost all (46 of 49) (p < 0.01). Reperfusion injury occurred in association with preoperative New York Heart Association (NYHA) III classification (p < 0.05), coronary endarterectomy (p < 0.01), long aortic clamping time (p < 0.01), and short postoperative survival (p < 0.05). CONCLUSIONS: Reperfusion injury was observed in one fourth of the deaths in association with MI. It occurred more often in patients with preoperative NYHA III symptoms and in those in whom endarterectomy was carried out and the anoxic time of the myocardium was longer. The shorter postoperative survival time indicates the lethal nature of this complication. PMID- 11016315 TI - Early and long-term results of coronary artery bypass grafting in dialysis patients. AB - BACKGROUND: Dialysis patients frequently present with debilitating coronary artery disease but are regarded as challenging patients for coronary artery bypass grafting. METHODS: The operative, early postoperative, and late results of 44 dialysis patients undergoing coronary artery bypass grafting from 1984 to 1997 were retrospectively reviewed. RESULTS: Compared with patients in The Society of Thoracic Surgeons database who underwent coronary artery bypass grafting, only cerebrovascular accident and postoperative cardiac arrest occurred more frequently in dialysis patients. However, 73% experienced some type of complication. Operative mortality was 11.4%. Decreased left ventricular ejection fraction and severe distal disease were predictive of increased operative mortality. New York Heart Association angina class fell from 2.8 to 1.5, and New York Heart Association congestive heart failure class fell from 2.6 to 1.8. Overall quality-of-life scores did not improve; however, walking distances remained consistently improved. Actuarial survival at 5 years was 32.0%+/-12.0%. Five-year survival was 0% for smokers and 83.6%+/-7.6% for nonsmokers (p = 0.0142). Causes of late death were myocardial infarction (4), sepsis (1), subdural hematoma (1), stroke (1), and unknown (6). CONCLUSIONS: Coronary artery bypass grafting should be avoided in dialysis patients with severe diffuse disease. A smoking history is associated with poor outcome. Coronary artery bypass grafting in dialysis patients is associated with a higher incidence of complications but can be performed with an acceptable operative mortality and is associated with good symptomatic relief of angina and heart failure. PMID- 11016316 TI - Right Y-graft, a new surgical technique using mammary arteries for total myocardial revascularization. AB - BACKGROUND: We report a new technique that consists of a right Y-graft using only skeletonized internal mammary arteries (IMA) for total arterial myocardial revascularization. METHODS: This technique consists of anastomosing the in situ left IMA (LIMA) and right IMA (RIMA) to the left anterior descending and obtuse marginal artery, via the transverse sinus, respectively. The distal free LIMA was anastomosed to the right coronary artery and afterwards in a Y fashion to the RIMA stem. Eleven patients with triple-vessel disease underwent coronary artery bypass grafting using this technique. Postoperatively and at follow-up all patients underwent color Doppler contrast-enhanced transthoracic echocardiography (TTE) before and after an adenosine provocation test. RESULTS: Overall, 33 IMA coronary anastomoses were made and 11 right Y-grafts were constructed. At 1 week after operation color Doppler contrast-enhanced TTE before and after the adenosine provocation test, respectively, showed an increase in LIMA stem diameter of 0.31 mm and in mean flow 62 mL/min. Coronary flow reserve (CFR) was 2+/-0.3. The increase in RIMA stem diameter was 0.2 mm and in mean flow was 121.7 mL/min. Coronary flow reserve was 2.5+/-0.4. Only 1 patient demonstrated an anomalous Doppler pattern, suggesting a partial Y-graft closure. CONCLUSIONS: Such a technique permits total myocardial revascularization using only mammary arteries and left ventricular perfusion from both IMAs simultaneously. The color Doppler contrast-enhanced TTE is a rapid, accurate, and noninvasive test allowing a good assessment of IMA patency. PMID- 11016317 TI - Basic fibroblast growth factor may improve devascularized sternal healing. AB - BACKGROUND: We have shown that a gelatin sheet incorporating basic fibroblast growth factor enhanced bone regeneration of the devascularized sternum. The purpose of this study was to determine if topical use of the gelatin sheet accelerated normal sternal regeneration and bone remodeling. METHODS: Thirty Wistar rats had median sternotomy and were divided into 3 groups: 10 had the bilateral internal thoracic arteries removed and basic fibroblast growth factor sheet applied on the sternum (group A), 10 had just the bilateral internal thoracic arteries removed (group B), and 10 had intact bilateral internal thoracic arteries (group C). RESULTS: Four weeks later the peristernal blood flow significantly increased and marked angiogenesis was seen around the sternum in group A. Histologically, the sternum was almost completely healed only in group A. In group A the bone mineral content was highest, but the bone mineral density was similar to that in other groups. The osteoclast index in group A was highest at the border zone of bone formation and remained high in regenerated bone. CONCLUSIONS: The basic fibroblast growth factor sheet offset sternal ischemia and accelerated normal sternal bone regeneration and remodeling, not only by callus formation but also by callus resorption. PMID- 11016318 TI - VEGF gene transfer mobilizes endothelial progenitor cells in patients with inoperable coronary disease. AB - BACKGROUND: Direct transfection of ischemic myocardium with naked plasmid DNA encoding for vascular endothelial growth factor-165 (VEGF165) has been shown to mobilize endothelial progenitor cells (EPCs). This study examined the kinetics of circulating EPCs isolated from peripheral blood mononuclear cells after gene transfer, and their role in neovascularization of ischemic myocardium. METHODS: The mononuclear cell population was isolated from peripheral venous blood samples of patients with functional class III or IV angina receiving intramyocardial VEGF165 gene transfer. Peripheral blood mononuclear cells were examined by an in vitro EPC culture assay and fluorescent-activated cell sorting. The data were compared with a control group consisting of patients who had undergone off-pump coronary artery bypass grafting without receiving gene transfer. RESULTS: Coinciding with a rise in VEGF levels, mobilization of EPCs increased significantly over base line for 9 weeks after the treatment (121+/-14 cells/mm2 versus 36.8+/-8 cells/mm2, p < 0.0005), followed by a subsequent decrease. Fluorescent-activated cell sorting analysis confirmed culture assay data, with a statistically significant rise in cells expressing vascular endothelial-cadherin, CD51/61 [alphavbeta3], CD62E [E-selectin], CD34, and KDR. The control group failed to show significant mobilization of EPCs. CONCLUSIONS: Mobilization of EPCs with resultant postnatal vasculogenesis, may play a role in revascularizing ischemic myocardium following human gene transfer with VEGF165. PMID- 11016319 TI - Robotic-assisted instruments enhance minimally invasive mitral valve surgery. AB - BACKGROUND: The potential for totally endoscopic mitral valve surgery has been advanced by the development of minimally invasive techniques. Recently surgical robots have offered instrument access through small ports, obviating the need for a significant thoracotomy. This study tested the hypothesis that a microsurgical robot with 5 degrees of freedom is capable of performing an endoscopic mitral valve replacement (MVR). METHODS: Dogs (n = 6) were placed on peripheral cardiopulmonary bypass; aortic occlusion was achieved with endoaortic clamping and transesophageal echocardiographic control. A small left seventh interspace "service entrance" incision was used to insert sutures, retractor blade, and valve prosthesis. Robotically controlled instruments included a thoracoscope and 5-mm needle holders. MVR was performed using an interrupted suture technique. RESULTS: Excellent visualization was achieved with the thoracoscope. Instrument setup required 25.8 minutes (range 12 to 37); valve replacement required 69.3+/ 5.39 minutes (range 48 to 78). MVR was accomplished with normal prosthetic valve function and without misplaced sutures or inadvertent injuries. CONCLUSIONS: This study demonstrates the feasibility of adjunctive use of robotic instrumentation for minimally invasive MVR. Clinical trials are indicated. PMID- 11016320 TI - A comparison of robot-assisted versus manually constructed endoscopic coronary anastomosis. AB - BACKGROUND: New technology has enabled surgeons to attempt totally endoscopic coronary artery bypass grafting. Our purpose was to compare three different techniques of totally endoscopic anastomosis using a porcine animal model. METHODS: Porcine hearts were excised and the right coronary artery was dissected free for use as an arterial graft. The hearts were placed in a human thoracic model and an endoscopic arterial anastomosis between the free right coronary artery and the left anterior descending coronary artery was performed using one of the following: (1) two-dimensional visualization with straight endoscopic instruments (n = 8); (2) three-dimensional head-mounted visualization with curved endoscopic instruments (n = 7); or (3) three-dimensional visualization with robotic telemanipulation (n = 8). Pathologic analysis of suture placement, vessel trauma, and patency was performed. Anastomoses were graded according to quality, ease, and patency using a seven-point Likert scale (1 = excellent, 7 = very poor). RESULTS: Endoscopic anastomotic ease and quality were significantly improved when three-dimensional visualization and curved endoscopic instruments were employed. Telemanipulation enhanced the process and provided the best operative results with regard to time required to construct the anastomosis, as well as ease and quality. CONCLUSIONS: Totally endoscopic anastomosis is feasible using currently available technology. Three-dimensional visualization and robotic telemanipulation significantly facilitate anastomosis construction and will likely benefit clinical operative outcome. PMID- 11016321 TI - Midterm angiographic assessment of coronary artery bypass grafting without cardiopulmonary bypass. AB - BACKGROUND: Coronary bypass surgery that provides good long-term graft patency can be performed on the beating heart as a viable alternative to conventional coronary artery bypass grafting (CABG). METHODS: From September 1993 to December 1996, 696 patients underwent CABG on the beating heart at the Kosuyolu Heart and Research Hospital in Istanbul. Among them, 70 patients were chosen randomly for angiographic assessment of off-pump coronary artery bypass grafting. RESULTS: The interval from operation to angiography varied from 24 to 61 months (mean, 36.1+/ 10.9 months). The patency rate of left internal mammary-left anterior descending artery anastomoses was 95.59% (patency achieved in 65 of 68 patients) and of vein grafts was 47.06% (patency achieved in 16 of 34 patients) (p < 0.0001). The patency rates of grafts anastomosed to the left anterior descending artery were significantly higher than the rates of the grafts anastomosed to the other coronary arteries (95.71% versus 45.45%, p < 0.0001). Multivariate analysis showed that graft type (p < 0.0001) and hyperlipidemia (p = 0.023) were significant predictors for graft occlusion. Left ventricular function improved significantly after CABG (p = 0.04). Reintervention (using percutaneous transluminal cardiac angioplasty) and reoperation rates were 0.97% and 1.4%, respectively. CONCLUSIONS: Off-pump coronary artery bypass grafting appears to produce midterm and long-term patency rates that are comparable to those of conventional techniques; that is especially true in cases of arterial conduits and of conduits anastomosed to the left anterior descending artery. PMID- 11016322 TI - Composite graft replacement of the aortic root after previous cardiac surgery: a 20-year experience. AB - BACKGROUND: An aging population and prolonged survival of patients after cardiac operations has meant that composite aortic root replacement after previous cardiac operation is being performed with increasing frequency. METHODS: From January 1979 to July 1999, 32 patients underwent "reoperative" composite replacement of the aortic root at our institution. Previous operations were 16 aortic valve replacement, 9 coronary artery bypass grafting, 5 repair aortic dissection, and 7 others. Indications for operation included ascending aortic aneurysm in 16 patients, ascending aortic dissections in 10 patients, and other in 6 patients. RESULTS: The unit elective mortality was 3 of 26 (11.5%). One surgeon's elective mortality was 1 of 22 (4.6%). The unit emergent mortality was 6 of 6 (100%). There has been one late death. Morbidity was low. CONCLUSIONS: Reoperative aortic root replacement is a technically demanding procedure, but expertise in the area achieves low elective mortality. Consideration should be given to aortic root replacement at the initial procedure. Close follow-up of postcardiac operation patients is necessary to proceed with elective aortic root replacement if indicated. Emergent presentation in the reoperative setting has a very poor prognosis. PMID- 11016323 TI - Safety of left innominate vein division during aortic arch surgery. AB - BACKGROUND: The surgical approach to the aortic arch via median sternotomy can be hindered by the left innominate vein (LIV). Retraction of the LIV may injure the vein. The safety of LIV ligation has been controversial. Opinion has also differed regarding whether a divided vein should be reanastomosed after arch replacement is completed. We report our experience with division and ligation of the LIV for improved aortic arch exposure and facilitated excision of mediastinal tumors. METHODS: From January 1996 to June 1998, the LIV was divided and ligated in 14 patients (8 men, 4 women) after consideration of local anatomy, adequacy of aortic arch exposure, level of distal aortic anastomosis, and in case of mediastinal tumors, extent of involvement of mediastinal structures. The LIV was divided between clamps, doubly ligated, and the ends oversewn. Patients were assessed at 1 month and at yearly intervals for upper extremity edema and neurologic symptoms. RESULTS: In 12 patients LIV division improved aortic arch access, and in 2 patients, it facilitated excision of mediastinal tumors. The mean age of patients was 56 years (range 22 to 80). Follow-up ranged from 1 week to 30 months. All patients had left upper extremity edema for 7 to 10 days, which resolved with arm elevation. One early patient required reexploration for bleeding from the LIV stump. One patient died because of multiorgan dysfunction. None had any residual left upper extremity edema or neurologic symptoms. CONCLUSIONS: We conclude that, although not uniformly or commonly necessary, division of the LIV can safely be utilized to facilitate aortic arch exposure without significant long-term morbidity. LIV reanastomosis is not necessary. PMID- 11016324 TI - Autologous smooth muscle cell transplantation improved heart function in dilated cardiomyopathy. AB - BACKGROUND: Transplantation of myocytes into scarred myocardium has been shown to inhibit ventricular remodeling and maintain myocardial contractility. However, the effect of cell transplantation on hearts with global rather than regional dysfunction is unknown. Therefore, we evaluated the effect of transplantation of autologous smooth muscle cells on the morphometry and function of dilated cardiomyopathic hearts. METHODS: Smooth muscle cells were isolated from the ductus deferens of 13-week-old BIO 53.58 hamsters with dilated cardiomyopathy, and cultured for 4 weeks before transplantation. Smooth muscle cells (4 x 10(6) cells) or culture medium were injected into 17-week-old animals in the transplantation and control groups (n = 12 each), respectively. Prelabeling of the smooth muscle cells with 5-bromo-2'-deoxyuridine was performed before transplantation in a group of transplanted hamsters. Another group (sham, n = 12) underwent the operation but did not receive an injection either of smooth muscle cells or of culture medium. Four weeks after transplantation, heart function was evaluated in a Langendorff preparation. RESULTS: Musclelike tissue, labeled with 5-bromo-2'-deoxyuridine, was found at the site of transplantation in the cell transplanted animals. The cell-transplanted hearts were smaller (p < 0.001), and had greater developed pressures and maximum rate of increase of left ventricular pressure (both p < 0.001) than control and sham hearts. Control hamsters injected with culture medium did not differ from sham-operated animals. CONCLUSIONS: Transplantation of autologous smooth muscle cells prevented cardiac dilatation and improved ventricular function in hamsters with dilated cardiomyopathy. PMID- 11016325 TI - Immediate coronary artery bypass surgery after platelet inhibition with eptifibatide: results from PURSUIT. Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrelin Therapy. AB - BACKGROUND: The platelet GP IIb/IIIa inhibitor eptifibatide improves outcomes in patients with acute coronary syndromes. Patients requiring emergent coronary artery bypass grafting, however, may be at increased risk for bleeding if exposed to eptifibatide. Data from the PURSUIT trial were reviewed to assess this risk in patients undergoing coronary surgery immediately after exposure to eptifibatide. METHODS: In PURSUIT, 10,948 patients who presented with non-ST segment elevation acute coronary syndromes were prospectively randomized to receive eptifibatide (180 microg/kg bolus plus 2 microg/kg/min infusion) or placebo. A total of 78 patients underwent immediate coronary artery bypass surgery within 2 hours of cessation of study drug (placebo, n = 46; eptifibatide, n = 32). Clinical outcome, bleeding, and transfusion requirements within this subset were examined. RESULTS: Major bleeding was not different between groups, occurring in 64% of patients receiving placebo and 63% of patients receiving eptifibatide. The incidence of blood transfusion was similar as well (57% vs 59%). Postoperative thrombocytopenia occurred less often after eptifibatide exposure. Perioperative myocardial infarction was significantly reduced in patients who received eptifibatide (46% vs 22%, p < 0.05). There was no difference in perioperative stroke (2.2% vs 6.3%) or mortality (6.3% vs 6.5%). CONCLUSIONS: Patients may safely undergo coronary artery bypass surgery within 2 hours of discontinuation of eptifibatide. Eptifibatide infusion in the immediate preoperative period had no adverse clinical effects, but did significantly decrease the incidence of perioperative myocardial infarction. Additionally, platelet counts after surgery were higher in the group of patients who received eptifibatide, perhaps indicative of a platelet-sparing effect during cardiopulmonary bypass. PMID- 11016326 TI - A randomized trial of antithrombin concentrate for treatment of heparin resistance. AB - BACKGROUND: Heparin resistance is an important clinical problem traditionally treated with additional heparin or fresh frozen plasma. We undertook a randomized clinical trial to determine if treatment with antithrombin (AT) concentrate is effective for treating this condition. METHODS: Patients requiring cardiopulmonary bypass who were considered to be heparin resistant (activated clotting time < 480 seconds after > 450 IU/kg heparin) were randomized to receive either 1000 U AT or additional heparin. RESULTS: AT concentrate was effective in 42 of 44 patients (96%) for immediately obtaining a therapeutic activated clotting time. This compared favorably to 28 of 41 patients (68%) treated with additional heparin (p = 0.001). All patients who failed heparin therapy were successfully treated with AT. The patients receiving AT required less time to obtain an adequate ACT but there was no difference in clinical outcomes among the groups. Study patients had deficient AT activity at baseline (56%+/-25%), which improved in those given AT concentrate (75%+/-31% versus 50%+/-23%, p < 0.0005). CONCLUSIONS: Heparin resistance is frequently associated with AT deficiency. Treating this deficiency with AT concentrate is more effective and faster for obtaining adequate anticoagulation than using additional heparin. PMID- 11016327 TI - Improvement by phosphoramidon of damaged endothelial function in porcine coronary artery. AB - BACKGROUND: The bradykinin (BK)-induced endothelium-dependent relaxation is impaired in the presence of elevated potassium concentration enhancing the vasospastic tendency of large coronary arteries. Inhibition of the angiotensin converting enzyme responsible for bradykinin degradation was found to enhance the endothelium-dependent relaxation by BK. The aim of the present study was to investigate the effect of phosphoramidon, known to inhibit a BK-metabolizing neutral endopeptidase enzyme, on relaxation of porcine-isolated coronary artery in depolarizing solution. METHODS: Endothelium intact porcine coronary artery rings were studied in organ chambers. The rings were isometrically contracted with potassium chloride (30 mmol/L) and the response to BK (1 to 1,000 nmol/L) induced relaxation was investigated in the presence of nitric oxide synthase inhibitor Nomega-nitro-L-arginine (300 micromol/L) alone and in combination with the cyclooxygenase inhibitor indomethacin (10 micromol/L), and that of the inhibitor of calcium-dependent potassium channels tetraethylammonium (7 mmol/L). Under these conditions, phosphoramidon (10 micromol/L), an inhibitor of a neutral endopeptidase enzyme (EC.3.4.24.11.), which is responsible for the degradation of BK, was used to enhance the endothelium-dependent relaxation. RESULTS: Phosphoramidon potentiated the maximum vasorelaxant effect of BK in Nomega-nitro L-arginine (control 26.6%+/-10.86% versus phosphoramidon 49.05%+/-4.52%; n = 6, p < 0.05) or in Nomega-nitro-L-arginine + indomethacin-pretreated rings (control 20.7%+/-9.92% versus phosphoramidon 42.0%+/-12.26%; n = 5, p < 0.05) and this increased vasodilation was not modified by tetraethylammonium. CONCLUSIONS: In the present study phosphoramidon potentiated the effect of BK in the absence of nitric oxide and prostaglandins in porcine-isolated coronary artery. This effect did not depend on tetraethylammonium-sensitive potassium channels. Phosphoramidon may be a useful pharmacologic tool for preserving the vasorelaxing capacity of coronary arteries after cardioplegia. PMID- 11016328 TI - Nitroglycerin is preferable to diltiazem for prevention of coronary bypass conduit spasm. AB - BACKGROUND: Diltiazem is widely used to prevent radial artery spasm after coronary bypass grafting (CABG). However, recent in vitro and in vivo studies have shown that nitroglycerin is a superior conduit vasodilator compared to diltiazem. A clinical comparison of these agents in patients undergoing CABG has not been previously performed. METHODS: One hundred sixty-one consecutive patients undergoing isolated CABG with the radial artery were prospectively randomized to 24-hour intravenous infusion of nitroglycerin or diltiazem followed by 6-month treatment with a daily dose of isosorbide mononitrate (n = 84) or diltiazem CD (n = 77). Analyses were performed on "intention-to-treat" basis. RESULTS: Crossovers because of low cardiac output, complete heart block, or sinus bradycardia occurred in 5 patients in the diltiazem group and none in the nitroglycerin group (p = 0.05). Operative mortality (nitroglycerin, 1.2% versus diltiazem, 1.3%), major morbidity (14% versus 16%), perioperative myocardial infarction (1.2% versus 0%), peak serum creatinine phosphokinase MB fraction levels (27 versus 21 U), intensive care unit stay (34+/-19 versus 38+/-30 hours) and total hospital length of stay (4.7+/-1.4 versus 4.7+/-1.3 days) were similar (p = not significant for all). Cardiac pacing was required more often in the diltiazem group (28% versus 13%, p = 0.01). Follow-up longer than 2 months was available in 145 patients (90%). Follow-up mortality (nitroglycerin, 1.2%; diltiazem, 1.3%), myocardial infarction (6%, versus 5%), and reintervention (8% versus 6%) rates and average angina class (1.3+/-0.7 versus 1.1+/-0.4) were similar (p = not significant for all). Thallium stress test obtained in 117 patients showed abnormal perfusion in the radial artery territory in only 4 patients (3%), 2 in each group (p = not significant). Treatment with diltiazem was more costly ($16,340 versus $1,096). CONCLUSIONS: Nitroglycerin is preferable to diltiazem for prevention of conduit spasm. Nitroglycerin is safe, effective, better tolerated, and less costly than diltiazem, and therefore, should be the agent of choice. PMID- 11016329 TI - Sialyl LewisX oligosaccharide preserves myocardial and endothelial function during cardioplegic ischemia. AB - BACKGROUND: Neutrophil adhesion to endothelium contributes to myocardial reperfusion injury after cardiac operation. Initial neutrophil-endothelial interactions involve selectins, which bind Sialyl-LewisX on neutrophils. Blockade of selectin-mediated neutrophil-endothelial interactions with CY-1503, a synthetic analogue of Sialyl-LewisX, might reduce reperfusion injury after myocardial ischemia. METHODS: The efficacy of CY-1503 to attenuate global myocardial reperfusion injury was assessed in isolated blood-perfused neonatal lamb hearts that had 2 hours of cold cardioplegic ischemia. CY-1503 (40 mg/L) or saline vehicle was added to blood perfusate before ischemia. Contractile function (developed pressure, dP/dt) and coronary vascular endothelial function (acetylcholine response) were assessed at base line and during reperfusion. Myocardial neutrophil accumulation was assessed by myeloperoxidase quantification. RESULTS: Compared to controls, treatment with CY-1503 improved recovery of all indices of contractile function, preserved coronary vascular endothelial function, and reduced myocardial neutrophil accumulation. CONCLUSIONS: In isolated neonatal lamb hearts that underwent hypothermic cardioplegic ischemia, CY-1503 administration reduced myocardial neutrophil accumulation and preserved endothelial and contractile function. Selectin blockade of leukocyte-endothelial interactions might attenuate reperfusion injury and enhance myocardial protection during cardiac surgical procedures. PMID- 11016330 TI - Preconditioning of swine heart with monophosphoryl lipid A improves myocardial preservation. AB - BACKGROUND: Ischemic preconditioning has been proven to be a powerful tool for myocardial protection in the setting of ischemia and reperfusion. A new drug to provide pharmacologic preconditioning, monophosphoryl lipid A (MLA), was administered 24 hours before an acute coronary occlusion in pigs to determine the effect on pharmacologic preconditioning. METHODS: Two studies were completed. In the first, swine were distributed into five groups: group I, control; group II,. aminoguanidine (AMG) (30 mg/kg), a selective inducible nitric oxide synthase (iNOS) blocker; group III, MLA (10 microg/kg); group IV, MLA (35 microg/kg); and group V, MLA and AMG (35 microg/kg and 30 mg/kg, respectively). Twenty-four hours after administration of the MLA, AMG, or both, regional left anterior descending coronary artery ischemia was induced for 15 minutes followed by one hour of global normothermic cardioplegic arrest and three hour reperfusion. Left ventricular function, tissue injury, and percentage of myocardial infarction were measured. Left ventricular myocardium in the left anterior descending coronary artery region was sampled for iNOS messenger RNA (mRNA) during ischemia and reperfusion. In the second study, pigs were sacrificed 0, 4, 6, 8, and 24 hrs after MLA/AMG administration for iNOS mRNA determination in nonischemic myocardium. RESULTS: Use of MLA significantly improved postischemic ventricular function, and reduced creatinine kinase release and percentage of infarction. Monophosphoryl lipid A induced expression of iNOS mRNA in nonischemic myocardium within four hours of administration which returned to base line by 24 hours. Normothermic regional ischemia then induced expression of iNOS mRNA, which returned to base line during reperfusion. Aminoguanidine completely abolished both MLA-induced and ischemia-induced iNOS mRNA and blocked the beneficial effects of MLA. CONCLUSIONS: Use of MLA can provide myocardial preservation through enhanced expression of iNOS mRNA. PMID- 11016331 TI - Myocardial protection using diadenosine tetraphosphate with pharmacological preconditioning. AB - BACKGROUND: We have reported a similar cardioprotective effect and mechanism of diadenosine tetraphosphate (AP4A) and ischemic preconditioning in rat hearts. In this study, the applicability of AP4A administration to cardiac surgery was tested by using a canine cardiopulmonary bypass model. METHODS: Hearts underwent 60 minutes of cardioplegic arrest (34 degrees C) by a single dose of cardioplegia. Cardioplegia contained either AP4A (40 micromol/L; n = 6) or saline (n = 6). Beagles were weaned from cardiopulmonary bypass 30 minutes after reperfusion, and left ventricular function was evaluated after another 30 minutes by using the cardiac loop analysis system. RESULTS: Administration of AP4A significantly improved the postischemic recovery of cardiac function and reduced the leakage of serum creatine kinase compared with saline. Systemic vascular resistance, mean aortic blood pressure, and the electrocardiographic indices were not significantly altered by AP4A administration. CONCLUSIONS: Administration of AP4A was cardioprotective without apparent adverse effects. Because the cardioprotective mechanism may be similar to that of ischemic preconditioning, the addition of AP4A into cardioplegia may be a novel safe method for clinical application of preconditioning cardioprotection. PMID- 11016332 TI - Minimally invasive esophagectomy. AB - BACKGROUND: Open esophagectomy can be associated with significant morbidity and delay return to routine activities. Minimally invasive surgery may lower the morbidity of esophagectomy but only a few small series have been published. METHODS: From August 1996 to September 1999, 77 patients underwent minimally invasive esophagectomy. Initially, esophagectomy was approached totally laparoscopically or with mini-thoracotomy; thoracoscopy subsequently replaced thoracotomy. RESULTS: Indications included esophageal carcinoma (n = 54), Barrett's high-grade dysplasia or carcinoma in situ (n = 17), and benign miscellaneous (n = 6). There were 50 men and 27 women with an average age of 66 years (range 30 to 94 years). Median operative time was 7.5 hours (4.5 hours with > 20 case experience). Median intensive care unit stay was 1 day (range 0 to 60 days); median length of stay was 7 days (range 4 to 73 days) with no operative or hospital mortalities. There were four nonemergent conversions to open esophagectomy; major and minor complication rates were 27% and 55%, respectively. CONCLUSIONS: Minimally invasive esophagectomy is technically feasible and safe in our center, which has extensive minimally invasive and open esophageal experience. Open surgery should remain the standard until future studies conclusively demonstrate advantages of minimally invasive approaches. PMID- 11016333 TI - Combined analysis of p53 and retinoblastoma protein expressions in esophageal cancer. AB - BACKGROUND: p53 gene mutation and abnormal p53 protein expression, also loss of the retinoblastoma gene and protein expression are frequently associated with esophageal squamous cell carcinoma (ESCC). Recently, the prognostic significance of the combined analysis of p53 protein and retinoblastoma protein (pRB) has been reported in non-small cell lung cancer. However, in ESCC, the prognostic significance of the combined analysis of these proteins remains unclear. In this study, we immunohistochemically analyzed the p53 protein and pRB expressions in surgically resected ESCC, and we evaluated the prognostic significance of the combination of these proteins. METHODS: We analyzed p53 protein and pRB expressions immunohistochemically in 191 surgically resected ESCC cases. Overexpression of p53 and loss of pRB were considered abnormal. RESULTS: Overexpression of p53 protein was detected in 79 patients (41%) and decreased pRB nuclear staining occurred in 82 (43%). The Kaplan-Meier survival curve showed that absence of pRB expression was significantly associated with shortened survival (p = 0.001), whereas expression of p53 was not significantly associated with survival. Moreover, p53 and pRB status individually were not independent prognostic factors in multivariate survival analysis. With respect to pRB and p53, the tumors could be grouped into four categories: p53-/pRB+ (31%); p53-/pRB- (27%); p53+/pRB+ (26%); and p53+/pRB- (16%). Favorable prognosis was observed in patients with p53-/pRB+ tumors. Multivariate analysis showed p53-/pRB+ status to be an independent prognostic factor. CONCLUSIONS: The combination of p53 protein loss and pRB expression was associated with good prognosis in patients with ESCC. PMID- 11016334 TI - Thoracoscopic completion thymectomy in refractory nonthymomatous myasthenia. AB - BACKGROUND: The aim of this study was to assess the efficacy of thoracoscopic completion thymectomy in patients with refractory nonthymomatous myasthenia. METHODS: Eight patients were operated upon after transcervical (n = 6) or transsternal (n = 2) thymectomy. The mean interval between operations was 129 months. Every patient was completely disabled despite treatment with large dosages of prednisone in combination with pyridostigmine (n = 5) or azathioprine (n = 3) and with repeated plasma exchanges. RESULTS: Gross (n = 5) or microscopic (n = 3) residual thymic tissue was found in all patients. There was no mortality, but morbidity included 2 patients with postoperative myasthenic crisis requiring reintubation and mechanical ventilation. The mean hospital stay was 4.75 days. The mean follow-up was 28.3 months. At the last follow-up, 6 patients had achieved symptomatic improvement as expressed by significant change in mean Osserman class (3.37 versus 2.12, p = 0.03), and prednisone dosage (43 versus 20 mg/d, p = 0.03). Conversely, there was no difference in dosage of pyridostigmine and azathioprine or in number of exchange cycles. CONCLUSIONS: Our results suggest that thoracoscopic completion thymectomy may be beneficial for selected patients with refractory nonthymomatous myasthenia. PMID- 11016335 TI - Comparison between closed drainage techniques for the treatment of postoperative mediastinitis. AB - BACKGROUND: It is not clear which closed drainage technique is preferred as initial therapy for mediastinitis as soon as it is detected after cardiac surgery. A comparison is made between a continuous irrigation system and vacuum drainage using redon catheters. METHODS: A retrospective cohort study of patients undergoing cardiac surgery between January 1, 1989 and January 1, 1997 was made. Patients who developed a deep surgical site infection at the sternotomy site and who were treated with one of the two closed drainage techniques were included. Patient characteristics and procedure-related variables were analyzed. Also, variables related to the drainage procedure were included. Outcome parameters were treatment failure, total hospital stay, postoperative hospital stay and in hospital mortality. RESULTS: The study population consisted of 11,488 patients, of whom 102 developed a deep surgical site infection (0.89%). The final study population consisted of 60 patients who fulfilled the inclusion criteria. From those, 29 were treated with continuous irrigation and 31 were treated with vacuum drainage. Both groups were comparable for patient characteristics and procedure related variables. Treatment failure was more than three times as likely in the continuous irrigation group (relative risk: 3.2, 95% confidence interval: 1.3 to 7.7). Also, postoperative (p = 0.03) and total hospital stay (p = 0.03) were significantly longer in the group treated with continuous irrigation (mean prolongation of 14 and 13 days, respectively). After correcting for confounding, using multivariate analysis, the treatment method employed was found to be an independent and statistically significant variable associated with treatment failure (p = 0.04). CONCLUSIONS: Closed drainage using vacuum-drainage system is the initial therapy of choice for patients with mediastinitis after cardiac surgery, because it is associated with significantly less treatment failure and a shorter stay in hospital. PMID- 11016336 TI - Inhibition of NF-kappaB sensitizes non-small cell lung cancer cells to chemotherapy-induced apoptosis. AB - BACKGROUND: Most non-small cell lung cancers (NSCLC) are chemoresistant. Identification and modulation of chemoresistance cell-signaling pathways may sensitize NSCLC to chemotherapy and improve patient outcome. The purpose of this study was to determine if chemotherapy induces nuclear factor-kappa B (NF-kappaB) activation in NSCLC in vitro and whether inhibition of NF-kappaB would sensitize tumor cells to undergo chemotherapy-induced apoptosis. METHODS: Non-small cell lung cancer cells were treated with gemcitabine, harvested, and nuclear extracts analyzed for NF-kappaB DNA binding by electrophoretic mobility shift assays. Additionally, NSCLC cells that stably expressed a plasmid encoding the superrepressor IkappaBalpha protein (H157I) or a vector control (H157V) were generated. These cells were then treated with gemcitabine and apoptosis determined by terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay. RESULTS: Chemotherapy induced NF-kappaB nuclear translocation and DNA binding in all NSCLC cell lines. H157I cells had enhanced cell death compared with H157V cells, suggesting that NF-kappaB is required for cell survival after chemotherapy. The observed cell death following the loss of NF-kappaB occurred by apoptosis. CONCLUSIONS: Inhibition of chemotherapy-induced NF-kappaB activation sensitizes NSCLC to chemotherapy-induced apoptosis in vitro. Novel treatment strategies for patients with advanced NSCLC may involve chemotherapy combined with inhibition of NF-kappaB-dependent cell-survival pathways. PMID- 11016337 TI - Pulmonary function after lobectomy: video-assisted thoracic surgery versus thoracotomy. AB - BACKGROUND: Whether video-assisted thoracic surgery (VATS) improves postoperative pulmonary function is still controversial. We compared postoperative pulmonary function after VATS lobectomy and standard lobectomy. METHODS: Eleven patients who had undergone standard lobectomy and 10 patients who had undergone VATS lobectomy were studied. Arterial blood gas analyses were performed on the 4th, 7th, and 14th postoperative days. Pulmonary function, including forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1.0), and peak flow rate (PFR) were measured on the 7th and 14th postoperative days (early phase), and approximately 1 year after surgery (late phase). RESULTS: Pulmonary function, as assessed with arterial oxygen partial pressure (PaO2) (p = 0.054), arterial oxygen saturation (O2SAT) (p = 0.063), FVC (p = 0.10), and FEV1.0 (p = 0.08), was better after VATS lobectomy than after thoracotomy on the 7th postoperative day. PFR was significantly better after VATS on both the 7th and 14th postoperative days (p = 0.008 and p = 0.03, respectively). CONCLUSIONS: VATS lobectomy had advantages on early postoperative pulmonary function. We conclude that VATS lobectomy is a beneficial alternative to standard thoracotomy, especially for patients with poor pulmonary reserve. PMID- 11016338 TI - Pneumoperitoneum after concomitant resection of the right middle and lower lobes (bilobectomy). AB - BACKGROUND: Removal of the right middle and lower lobes often leaves a pleural space problem that can cause prolonged air leaks. METHODS: A single surgeon prospectively randomized 16 patients who underwent bilobectomy. Eight patients had 1200 mL of air injected under the right hemidiaphragm after bilobectomy and 8 did not. The air was injected through a small transdiaphragmatic opening made in the right hemidiaphragm at the time of pulmonary resection. RESULTS: The age of the patients, preoperative pulmonary function, preoperative comorbidities, indications for surgery, and final pathology were not significantly different between the two groups. On postoperative day #1, a pneumothorax was present in 1 patient (13%) in the pneumoperitoneum group (P group) and in 4 patients (50%) in the nonpneumoperitoneum group (N-P group). On postoperative day 1, an air leak was present in 1 patient (13%) in the P group and 5 patients (63%) in the N-P group (p < 0.001). By the third postoperative day, no patient in the P group had an air leak; however, a leak was present in 4 patients (50%) in the N-P group (p < 0.001). Median hospital stay in the P group was 4 days (range, 3 to 6 days), compared with 6 days (range, 4 to 8 days) in the N-P group (p < 0.001). Three patients in the N-P group were sent home with a Heimlich valve. There was no operative mortality and no complications from the pneumoperitoneum. CONCLUSIONS: We conclude that pneumoperitoneum after bilobectomy is safe and easy to do. It decreases the incidence of air leaks and of pneumothoraces and shortens hospital stay without increasing morbidity. We recommend pneumoperitoneum after bilobectomy at the time of thoracotomy, especially if there are residual small air leaks that cannot be sealed before chest closure. PMID- 11016339 TI - Reduction pneumoplasty versus respiratory rehabilitation in severe emphysema: a randomized study. Pulmonary Emphysema Research Group. AB - BACKGROUND: The purpose of the study was to determine in a prospective randomized trial the independent short-term physiologic impact of reduction pneumoplasty (RP) on respiratory rehabilitation (RR). METHODS: Sixty patients eligible for RP were randomly selected by computer to receive either RP (n = 30) or comprehensive RR (n = 30). Pulmonary function tests, analysis of blood gas levels, measurement of respiratory muscle strength (maximal inspiratory and expiratory pressures), 6 minute walk test (6MWT), and incremental treadmill test (ITT), were performed at baseline and at 3 and 6 months. RESULTS: Two treatment-related deaths occurred after RP and one after RR. At 6 months dyspnea index, maximal inspiratory pressure, 6MWT, ITT, and PaO2 were significantly improved in both groups whereas forced expiratory volume in 1 second and residual volume were significantly improved only in the surgical arm. In addition at 6 months, dyspnea index, 6MWT, maximal ITT, and PaO2 improved significantly more after RP than after RR. CONCLUSIONS: In our study short-term improvements in dyspnea index, oxygenation, inspiratory muscle strength, and exercise capacity occurred after either RP and RR. However dyspnea index, PaO2, and exercise capacity improved more after RP than after RR whereas pulmonary function improved only after RP. PMID- 11016340 TI - Main pulmonary artery laceration after blunt trauma: accurate preoperative diagnosis. AB - Blunt chest trauma is associated with a variety of lethal injuries, many of which are responsible for prehospital mortality. Major intrathoracic vascular injury accounts for a vast majority of these fatal injuries. Patients surviving after main pulmonary artery injury are rare. We present the case of a patient who sustained a main pulmonary artery laceration as a result of a blunt motor vehicle crash. He was diagnosed accurately by computed tomography and underwent successful repair. PMID- 11016341 TI - Mediastinal dumbbell angiolipoma. AB - Nonneurogenic dumbbell tumors are rare. This report describes the case of a 46 year-old woman with a symptomatic mediastinal dumbbell angiolipoma. The tumor was successfully resected using a single-stage procedure, combining a posterior microneurosurgical and thoracoscopic approach. The patient made an uneventful recovery and the neurologic symptoms improved immediately. PMID- 11016342 TI - Left ventricular reduction in a Jehovah's Witness. AB - For Jehovah's Witnesses with severe heart failure, left ventricular reduction surgery may be a satisfactory alternative to cardiac transplantation. Compared with transplantation, left ventricular reduction surgery can involve less blood loss thus decreasing the need for blood-volume replacement. More importantly, left ventricular reduction surgery obviates the need for a donor organ. PMID- 11016343 TI - Dramatic improvement of LV function after coronary sinus thromboembolectomy. AB - We present the case of a 79-year-old man suffering from chronic atrial fibrillation, severe left ventricular dysfunction, massive right atrial thrombosis, and pulmonary hypertension. Complete coronary sinus thrombosis was found incidentally during preoperative screening. Successful coronary sinus, right atrial, and pulmonary operative embolectomy was followed soon after by a dramatic improvement of cardiac performance; the patient's left ventricular function recovery, in particular, suggests that cardiac venous system played an important role in the genesis of myocardial impairment. PMID- 11016344 TI - Reoperative revascularization of a left internal mammary artery ostial stenosis. AB - A severe ostial stenosis of the left internal mammary artery graft was responsible for unstable angina in a patient with a previous coronary artery bypass graft. Successful revascularization of the lesion was achieved with a subclavian artery-to-left internal mammary artery bypass using a saphenous vein conduit. This procedure was performed through a left thoracotomy incision to avoid potential hazards of a redo median sternotomy. PMID- 11016345 TI - Aortobronchial fistula late after transverse arch replacement. AB - We report an unusual case of aortobronchial fistula late after transverse arch replacement caused by the remnant of a temporary bypass near the ascending aorta. In reconstructive surgery of the ascending aorta, antegrade perfusion is preferably performed through a side branch after completion of the distal anastomosis by some surgeons. This report suggests possible risk of a serious late complication unless the side branch is placed and tailored properly. PMID- 11016346 TI - Endarteritis and false aneurysm complicating aortic coarctation. AB - We report a tricky case of endocarditis because of the localization, aortic coarctation, and the pathogenic bacteria Actinobacillus actinomycetemcomitans. Furthermore, we underline the leading role of transesophageal echocardiography in the diagnosis of aortic endarteritis. First, aortitis was treated with antibiotics and, second, successfully operated on. PMID- 11016347 TI - Right ventricle lipoma with pseudoaneurysmatic appearance. AB - The case of a 78-year-old female patient, with a large lipoma (13 x 17 x 10 cm) of the right ventricle, appearing pseudoaneurysmatic, is presented. Radical resection left a considerable right ventricle wall and interventricular septum defect which was reconstructed with a bovine pericardium patch. PMID- 11016348 TI - L-->R shunt: a serious consequence of TAPVC repair without ligation of vertical vein. AB - It has been suggested that concomitant ligation of the vertical vein (VV) is not necessary in the repair of total anomalous pulmonary venous connection. The patency of the VV is desirable in the presence of noncompliant left heart chambers that may not be able to accommodate acute increases in pulmonary blood flow, leading to hemodynamic instability after repair. Complete cessation of flow through the previously patent VV has been observed, obviating the need for a second-stage operation. We report 2 infants who were operated using this strategy, in which the VV continued to function as a conduit for a significant left to right shunt. PMID- 11016349 TI - Division of venous collateral after Glenn shunt by minimally invasive surgery. AB - An infant with cyanotic heart disease that was palliated with a bidirectional cavopulmonary shunt developed progressive cyanosis 3 months after the surgical procedure. A large hemizygous vein was found to be decompressing the bidirectional Glenn shunt from the left innominate vein and was ligated using video-assisted thoracoscopic surgery. The unusually rapid appearance of the shunt, and the excellent outcome associated with the video-assisted thoracoscopic surgery procedure are discussed. PMID- 11016350 TI - Hemangioma of the right ventricular outflow tract. AB - Obstruction of the right ventricular outflow tract by a primary cardiac tumor is rare. Six cases of right ventricular outflow tract obstruction by a primary cardiac hemangioma have been reported; all but one were detected before the age of 25 years. In this report, we review the literature and describe what we believe to be only the second reported case of right ventricular outflow tract obstruction produced by a cardiac hemangioma that presented in late adulthood. PMID- 11016351 TI - Rare presentation of cardiac hemangiomas. AB - Cardiac hemangiomas are exceptionally rare tumors with an incidence of 1% to 2% of all detected benign heart neoplasms. The clinical appearance of the tumor varies considerably and may mimic other pathological findings of definite heart structures. We report two cases of cardiac hemangiomas presenting with an unusual location and clinical course. PMID- 11016352 TI - Missiles in the heart causing coronary artery disease 44 years after injury. AB - We present a case of two missiles in the heart causing coronary artery disease with first onset 44 years after the injury. The missiles were close to the right coronary artery and the left anterior descending coronary artery. The missiles initiated local arteriosclerosis in these vessels, with an extremely stretched time frame from injury to the onset of symptoms for coronary artery disease. PMID- 11016353 TI - Changes in graft flow pattern from the descending aorta due to intraaortic balloon pump. AB - During surgery for lung cancer in a patient who had undergone coronary artery bypass grafting through the descending aorta by left thoracotomy, we measured graft bypass blood flow from the descending aorta under intraaortic balloon pump (IABP) assistance. Under IABP assistance, the diastolic waveform changed to a spiky pattern with a sharp drop in blood flow of approximately 16% compared to that without IABP assistance. We report changes in graft flow pattern during IABP assistance when the graft is placed from the descending aorta. PMID- 11016354 TI - Assessment of thoracoscopic sympathectomy by infrared thermal imaging. PMID- 11016355 TI - Transcervical-transtracheal endoluminal repair of membranous tracheal disruptions. AB - Tracheal lacerations are rare and potentially hazardous complications of tracheal intubation. Surgical repair is the treatment of choice of tracheal injuries although nonoperative management is occasionally appropriate for well-selected patients. We describe our personal technique of anterior transcervical transtracheal endoluminal suture of iatrogenic lacerations of the membranous trachea and our results with this approach in 8 patients. This method is less invasive than conventional cervical or transthoracic approaches. PMID- 11016356 TI - Wedge carinal resection for closure of the main bronchus after pneumonectomy. AB - Closure of the bronchial stump after pneumonectomy can be challenging. Some special situations such as tumor extension, technical pitfalls, or poor tissue quality of the bronchial stump may preclude safe closure of the airway with standard techniques. We describe here a technique of wedge carinal resection that provides the surgeon an alternative whenever the standard closure of the stump is inapplicable. This technique has been successfully used in a series of 4 patients. PMID- 11016357 TI - A circuit to perform a combined ultrafiltration procedure in pediatric open heart surgery. AB - Cardiopulmonary bypass (CPB) induces an increased capillary permeability and tissues water content due to hemodilution and the inflammatory response, resulting in organ dysfunction. The reduction of the water accumulation and inflammatory response can be achieved by employing ultrafiltration during CPB. Recently we developed a simple CPB circuit for ultrafiltration using the aortic venting tube as an inlet line. Such a technique offers the advantages of performing a combined ultrafiltration procedure and eliminating the danger of air embolism. We employed this circuit in 12 consecutive pediatric patients undergoing open heart surgery. PMID- 11016358 TI - Congenital foramen of the left pericardium. AB - Congenital foramen of the left parietal pericardium is uncommon. The condition has the potential to cause angina pectoris, myocardial infarction, or even death. Forty-three confirmed cases have been retrieved from the English language literature, and a case report of "life-threatening" herniation of the heart is here added. The diagnosis, made at a mean age of 20 years (range 2 to 48), was five times more common in men. In 5 fatal cases, the heart had become incarcerated. In the remainder of cases, one-third were asymptomatic, and two thirds suffered a chest complaint that prompted diagnosis. Chest discomfort, dyspnea, and syncope were the most common symptoms. The most common finding at surgery, which 34 patients underwent, was a foramen at the base of the heart through which the left atrial appendage had herniated. In eight instances, the rim of the defect lay upon and compressed the coronary circulation. Measures to remedy the disorder have included a variety of operations, some to enlarge the defect, others to close it, amputation of the atrial appendage, and, in two cases, myocardial revascularization. Surgery is appropriate in the majority of symptomatic patients and in all who are at risk for ventricular herniation. PMID- 11016359 TI - As originally published in 1994: Simplified, standardized technique for cervical esophagogastric anastomosis. Updated in 2000. AB - Using transhiatal esophagectomy, the ease of performing the cervical esophagogastric anastomosis varies greatly depending on neck size, length of mobilized stomach, and adhesions from previous neck operations. We therefore have developed a technique, used in 45 consecutive patients undergoing transhiatal esophagectomy, that has simplified the technical performance of cervical esophagogastric anastomosis. PMID- 11016360 TI - Volume-outcome relationship in thoracic surgery. PMID- 11016361 TI - pH-stat strategies protect 60-minute central nervous system ischemia at 29.5 degrees C. PMID- 11016362 TI - Cardiac and pericardiac echinococcosis. PMID- 11016363 TI - How to avoid problems with Freestyle valve implantation. PMID- 11016364 TI - The ITA pedicle: a "protecting cloak" for the graft of choice. PMID- 11016365 TI - Emergence of a new direction in our specialty: catheter-assisted cardiac surgery. PMID- 11016366 TI - A review of 1,582 consecutive Octopus off-pump coronary bypass patients. AB - BACKGROUND: Off-pump coronary bypass may provide a safer form of surgical revascularization by avoiding the unwanted complications of cardiopulmonary bypass, particularly in the increasingly complex patients being referred for operation. This study reviews the entire experience of the Medtronic Octopus System (Medtronic, Minneapolis, MN) for beating heart bypass from 7 surgeons. Demographics, operative procedures, early outcomes, and trends in usage were examined. METHODS: Patients were selected for off-pump procedures by the individual surgeons. Data were entered prospectively into locally maintained databases and then collected for collation and analysis. RESULTS: A total of 1,582 consecutive Octopus patients were entered, representing the entire Octopus experience of each surgeon. Proportions of off-pump procedures relative to standard bypass increased over time, as did the percentage of patients receiving three or more grafts, 24.6% in 1997 and 55.9% in 1999. A total of 3,653 anastomoses were performed, 1,905 to the left anterior descending system, 837 to the circumflex distribution, and 911 to the right coronary territory. Morbidity was low. Few patients required conversion to cardiopulmonary bypass (2.6%; 0.2% urgently). Permanent stroke occurred in 0.6% and myocardial infarction in 1.2%. Operative mortality was 1%. CONCLUSIONS: Octopus off-pump bypass was demonstrated to be a safe procedure with widening applicability. With experience surgeons tend to apply the system to increasing proportions of their patients and are able to revascularize all coronary territories. PMID- 11016367 TI - Is low ejection fraction safe for off-pump coronary bypass operation? AB - BACKGROUND: Does the manipulation of the heart during off-pump coronary artery bypass (OPCAB) procedure further compromise the hemodynamic stability of a patient with depressed left ventricular function compared with the conventional coronary artery bypass (CCAB) approach? Does this manipulation induce a more dramatic hypoperfused state that may contribute to an increase in the incidence of related complications or mortality? This retrospective review of data attempted to answer the above concern. METHODS: Between January 1, 1998, and June 30, 1999, 177 patients with ejection fractions of 30% or less underwent full sternotomy coronary artery bypass grafting at our institution. Of these patients, 45 underwent OPCAB procedures and 132 patients underwent CCAB. Pre-, intra-, and postoperative variables as identified by The Society of Thoracic Surgeons National Cardiac Surgery Database were compared using univariate and logistical regression analysis. RESULTS: Despite recognized hemodynamic derangement during cardiac displacement, these groups of OPCAB patients appeared to tolerate the procedure well. Univariate analysis of cardiac enzyme leak and blood loss was statistically significant in the OPCAB patients. Utilizing regression analysis, cardiopulmonary bypass was the only predictor for all postoperative complications. CONCLUSIONS: Multivessel coronary artery bypass utilizing the OPCAB approach in patients with depressed left ventricular function of equal to or less than 30% is appropriate and applicable. Analysis of CCAB and OPCAB variables was nonsignificant except for operative and postoperative blood loss and peak cardiac enzyme leak. Attention to intraoperative detail and hemodynamic management could be credited for the success with OPCAB. PMID- 11016368 TI - Off-pump redo coronary artery bypass grafting. AB - BACKGROUND: Conventional redo coronary artery bypass grafting is associated with significant morbidity. The danger of reoperation is mainly in reopening the sternum and in the manipulation of the heart and the old grafts. Therefore, off pump redo coronary artery bypass grafting with a patient-specific approach in selected cases seems an ideal technique. METHODS: Between October 1995 to September 1999, 50 patients with mean age of 61.8+/-8 years underwent reoperative coronary artery bypass grafting without cardiopulmonary bypass. Isolated left internal mammary artery (LIMA) to left anterior descending artery (LAD) anastomosis was carried out in 25 cases through left anterior minithoracotomy. In 1 patient LIMA was grafted on a previous vein graft to LAD, which was critically stenosed proximally but distal anastomosis was patent. In another case LIMA was grafted to Ramus intermedius branch. Midsternotomy approach was used to carry out LAD and right coronary artery grafting in 21 cases. In 2 patients a posterolateral thoracotomy approach was used to bypass obtuse marginal branches without cardiopulmonary bypass; in these cases proximal anastomosis was performed on the descending aorta. RESULTS: Mortality rate was 4% (2 deaths). Two patients sustained perioperative myocardial infarction. No patient was reexplored for hemorrhage and 38 patients did not require homologous blood transfusion. Sixteen patients underwent check angiogram and all of them were found to have patent redo grafts. Cardiac recovery room stay was 22+/-7 hours and hospital stay 5+/-2 days. CONCLUSIONS: In selected patients, reoperative coronary artery bypass grafting can be performed without cardiopulmonary bypass with a low perioperative morbidity and mortality and satisfactory graft patency. PMID- 11016369 TI - Off-pump coronary operation at the Favaloro Foundation: results in 264 patients. AB - BACKGROUND: Our objective was to determine whether complete myocardial revascularization "off pump" was feasible while maintaining the efficacy of conventional surgical techniques. METHODS: 264 patients were operated on between March 15, 1998, and August 26, 1999. A total of 218 (82.6%) were men and the average age was 61.4 years (range 28 to 87 years). Left main occlusions were present in 10.2% and 84% had multiple vessel disease. A total of 628 grafts were implanted, an average of 2.4 grafts per patient. Total arterial revascularization was achieved in 81.5% using both internal mammary arteries and the radial artery, one or more venous grafts in 18.1%, and venous alone in 0.3%. RESULTS: Surgical mortality was 2.2% and perioperative myocardial infarction was 3%. There were no neurologic events in this group of patients. In the first 73 patients coronary angiography control was performed before discharge and all grafts were patent. Angioplasty was carried out in 2 patients (0.76%) and conversion of procedure in 4 (1.5%). Follow-up time was 4 to 14 months. CONCLUSIONS: This procedure enabled revascularization of all areas of the heart, usually with total arterial revascularization, and excellent patency rates. The morbidity and mortality observed was similar to conventional surgery. The incidence of secondary complications (bleeding, need for transfusion, prolonged mechanical ventilation, or neurologic events) was lower. PMID- 11016370 TI - Off-pump surgery for anterior vessels in patients with severe dysfunction of the left ventricle. PMID- 11016371 TI - Surgical strategies in patients at high risk for stroke undergoing coronary artery bypass grafting. AB - BACKGROUND: Perioperative stroke represents one of the major complications following coronary artery bypass grafting (CABG). The present study was designed to evaluate the use of an individualized surgical approach for reducing neurological injury in patients undergoing CABG at high risk of stroke from aortic atherosclerosis or carotid disease. METHODS: Between January 1993 and June 1999, 6,138 patients undergoing elective CABG were evaluated by intraoperative transesophageal echocardiography. Patients were screened preoperatively for internal carotid artery disease. Based on the intraoperative transesophageal echocardiography findings the surgical technique was individualized: hypothermic circulatory arrest with aortic atherectomy, CABG combined with transmyocardial laser revascularization on the beating heart, off-pump CABG by midsternotomy, ministernotomy, minimally invasive direct CABG, hybrid procedure, and so on. Patients were divided into four groups: a low-risk group (no significant aortic or carotid disease); an aortic atheromatous disease group (A.ATH); a carotid disease group (CD); and a carotid disease combined with aortic atheromatous disease group (CD + A.ATH). RESULTS: The incidence of stroke in the low-risk group (n = 5,043) was 0.92% compared with 0.96% in the A.ATH group (n = 918). In the CD group (n = 166) the incidence of stroke was 0.6% whereas it was 0% in the CD + A.ATH group (n = 11). CONCLUSIONS: Preoperative and intraoperative screening can detect extensive atherosclerosis of the proximal aorta and internal carotid artery. Selective use of surgical techniques in this group of high-risk patients can prevent adverse neurologic sequelae while achieving complete myocardial revascularization. PMID- 11016372 TI - An automated interrupted suturing device for coronary artery bypass grafting: automated coronary anastomosis. AB - BACKGROUND: The purpose of this study was to have a preliminary assessment of the safety and efficacy of an automated vascular suturing device. METHODS: The device (Heartflo, Perclose/Abbott Labs, Redwood City, CA), which delivers 10 interrupted 7-0 polypropylene sutures between side-to-side arteriotomies, was evaluated in animals (8 Yorkshire pigs). RESULTS: Tissue edge capture and quality of anastomosis were highly rated. Time of anastomoses averaged 22 minutes. This time was prolonged primarily due to suture management, tying of interrupted sutures, and learning curve effects. Six of the anastomoses were hemostatic and two required an additional stitch each. Angiography and histology of the anastomosis confirmed patency and quality of the anastomosis. CONCLUSIONS: Our preliminary results indicate that the Heartflo automated anastomotic device is safe and effective. Preclinical and clinical studies to validate its acute and long-term effectiveness will commence shortly. PMID- 11016373 TI - Coronary artery bypass grafting on the beating heart evaluated with integrated backscatter. AB - BACKGROUND: In beating heart coronary artery bypass grafting (CABG) the effect of ischemic insult during coronary occlusion could not be evaluated immediately. Using transesophageal echocardiography, myocardial performance can be evaluated with analysis of integrated backscatter. METHODS: In 15 beating heart CABGs, cyclic variation (CV) of integrated backscatter of the anterior wall before, during, and after the left internal thoracic artery to left anterior descending (LAD) branch anastomosis was measured with transesophageal echocardiography. The patients were divided into two groups according to collateral vessels status (good collateral group n = 6, poor collateral group n = 9). RESULTS: In all patients, CV increased significantly after revascularization (8.56+/-2.50 to 11.47+/-3.32 dB, p < 0.0001). During LAD occlusion, significant decrease in CV was found in patients who had poor collateral arteries. At 15 minutes of LAD occlusion, CV decreased from the preocclusion value of 7.51+/-2.21 to 3.23+/-4.03 dB (p < 0.01). CONCLUSIONS: Measurement of CV can detect the ischemic insult during coronary occlusion and the effect of revascularization in beating heart CABG. PMID- 11016374 TI - Predictors of outcome in a multicenter port-access valve registry. AB - BACKGROUND: The aim of this study was to examine the predictors of outcome in patients undergoing isolated valve operation using port-access techniques. METHODS: Logistic regression analysis was performed in a prospective, multi institutional registry of patients undergoing isolated aortic valve replacement (AVR, n = 252), mitral repair (MVP, n = 491), or mitral replacement (MVR, n = 568) using port-access techniques from 1997 to 1999. RESULTS: Endoaortic balloon occlusion was used in 2% (AVR), 93% (MVP), and 90% (MVR) of cases. Conversion to full sternotomy occurred in 3.8% of all cases. For all patients, early mortality was 50 of 1,311 (3.8%) and onset of new atrial fibrillation occurred in 140 of 1,311 (11%) patients. The determinants of 30-day mortality were redo, age, and MVR or AVR. The determinants of reoperation for bleeding were age, reoperation, and MVR. Age was a predictor for stroke, and age and low or medium volume center were predictors of new atrial fibrillation. CONCLUSIONS: Excellent short-term results can be obtained using port-access techniques in isolated mitral or aortic valve operations. Patient outcome is not related to institutional case volume, and the primary determinants of outcome after port-access valve procedures are generally patient-related factors. PMID- 11016375 TI - Robotic-enhanced arterial revascularization for multivessel coronary artery disease. AB - BACKGROUND: A tendency to reduce operative trauma is determining the evolution of cardiac surgical techniques lately. The introduction of robotic-enhanced endoscopic systems enables surgeons to perform arterial revascularization for multivessel disease without sternotomy. METHODS: From May 1999,17 (4 women, 13 men; median age 63+/-7.4 years) patients with multivessel coronary artery disease were treated surgically using arterial revascularization by means of bilateral internal mammary arteries. Both arteries were harvested endoscopically using the da Vinci system (Intuitive Surgical, Mountain View, CA). These vessels were anastomosed using the "Dresden technique." RESULTS: Survival was 100%. Mean duration of the operation was 255+/-40.4 minutes. Bilateral internal mammary artery harvesting took 88.5+/-15.9 minutes; cross-clamp time was 36+/-8.7 minutes. An average of 2.06 anastomoses were performed per operation. Postoperatively, patients remained in the intensive care unit for 21+/-13 hours. One patient (5.8%) needed reexploration due to bleeding. CONCLUSIONS: The robotic surgical system introduces a new treatment of coronary artery disease to surgical practice, and enables arterial revascularization with distinctly reduced surgical trauma. PMID- 11016376 TI - Minimally invasive direct coronary artery bypass grafting: intermediate-term results. AB - BACKGROUND: Intermediate- and long-term clinical outcome and graft patency in minimally invasive direct coronary artery bypass grafting (MIDCABG) procedures remain a concern. METHODS: Over a 13-month period, 66 MIDCABG procedures were performed utilizing robotic-assisted internal mammary artery (IMA) harvesting and direct CABG through a 5-cm thoracotomy without cardiopulmonary bypass. Clinical follow-up was obtained on all patients. Graft patency was assessed in 61 of 66 consecutive patients 6 months (range 2 to 15 months) postoperatively. Group I consisted of 45 patients who underwent IMA angiography and group II consisted of 16 patients who underwent both a nuclear stress test and transthoracic Doppler examination. All group II patients had abnormal preoperative nuclear stress tests for comparison. RESULTS: To date, all 66 patients are alive. Graft patency rates in the two groups were 97.8% (45 of 46 grafts in 45 patients) in group I and 100% (15 of 15 grafts), with one indeterminate study, in group II. The overall patency rate for the entire study group was 98.3% (60 of 61 grafts). Sixty-two of 66 (93.9%) patients were able to return to their normal level of activity within 3 weeks. CONCLUSIONS: This study demonstrates that the MIDCABG with thoracoscopic IMA harvesting can achieve effective intermediate-term revascularization and an acceptable clinical outcome. PMID- 11016377 TI - Angiographic evaluation of graft patency in minimally invasive direct coronary artery bypass grafting. AB - BACKGROUND: The objective of this report is to describe our experience using minimally invasive direct coronary artery bypass grafting and to evaluate angiographic patency of anastomoses. METHODS: A total of 120 patients (86 men) were operated on, with ages ranging from 30 to 83 years (mean = 61.2 years). Two access routes were used: for single left anterior descending coronary artery lesions an 8 cm anterior minithoracotomy was performed at the fourth left intercostal space. Extracorporeal circulation was not used. In the last 82 patients a restraining device was used for the regional reduction of heart beats. Coronary cineangiography was carried out between postoperative days 1 and 3 in 84 (70%) patients. Anastomoses were graded: grade A, no blocks; grade B, blocks of more than 50%; grade C, occlusion. This evaluation was performed for two different periods: in the first period a restraining device was not used and in the second period a restraining device was used. RESULTS: In the first study period (38 anastomoses) coronary cineangiography showed grade A, 79%, grade B, 5.2%, and grade C, 15.8%. In the second study period (62 anastomoses), angiography showed grade A, 90.4%, grade B, 6.4%, and grade C, 3.2%. Early mortality was 1.6%. CONCLUSIONS: Minimally invasive coronary artery bypass grafting is a good alternative for some groups of patients. Anastomotic results seem to be better when a restraining device is used. PMID- 11016378 TI - Off-pump coronary artery bypass grafting: initial experience at one community hospital. AB - BACKGROUND: This study reports one cardiac surgical center's experience with off pump coronary artery bypass (OPCAB) and compares clinical risk factors and outcomes with a group of patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass at the same institution. METHODS: Data on preoperative risk factors, intraoperative clinical markers, and postoperative outcomes were collected prospectively on all patients undergoing cardiac surgical procedures at our institution. From January 1, 1999, through October 7, 1999, 332 patients underwent OPCAB procedures at our institution. This group was compared with 445 consecutive patients undergoing CABG at the same institution during the period of January 1, 1998, through November 30, 1998. RESULTS: The two groups were similar with respect to preoperative clinical risk factors. Intraoperative data showed OPCAB patients tended to have fewer grafts performed and had a lower frequency of multiple grafts to obtuse marginal vessels. Outcomes showed no differences in the incidence of perioperative stroke, mediastinitis, reexploration for bleeding, pulmonary complications, new renal failure, postoperative atrial fibrillation, or transfusion of blood products. Patients in the OPCAB group had fewer perioperative myocardial infarctions and lower incidence of postoperative low cardiac output syndrome. A higher percentage of OPCAB patients had surgical lengths of stay of 5 days or less. The OPCAB group tended to have a lower in-hospital mortality rate but this difference did not reach statistical significance. CONCLUSIONS: Off-pump coronary artery bypass grafting with revascularization of all coronary artery segments is a safe and effective procedure that can be performed with equal or improved outcomes and shorter surgical lengths of stay compared with CABG with cardiopulmonary bypass. PMID- 11016379 TI - Mitral valvuloplasty with sutures used for aortic prosthesis implantation. AB - BACKGROUND: The purpose of this study was to analyze surgical treatment for aortic valve lesions with coexisting mitral regurgitation (MR). METHODS: Seventy five patients were divided into two groups according to intensity of MR (group 1, MR less than or equal to II; group 2, MR greater than or equal to II/III). There were two control groups (control 1, only patients with implantation of aortic valve; control 2, patients with implantation of both aortic and mitral prostheses). During implantation of a mechanical aortic prosthesis, the same suture for fixation of a mechanical prosthesis and for suspension of mitral commissural regions and lifting the base of anterior mitral cusp was used. RESULTS: In all patients, no early death occurred. There were two late deaths, one due to endocarditis, and the other to heart failure. All patients from both groups had decreased MR. Selected echocardiographic parameters improved: end diastolic and end systolic diameter and ejection fraction in group 2 improved in proportion to patients in whom mitral valves were implanted (control 2). CONCLUSIONS: Simultaneous suspension of the mitral commissure area during mechanical aortic prosthesis implantation reduces associated MR. This technique seems to be efficient during implantation of aortic prostheses in patients with coexisting MR. PMID- 11016380 TI - Minimal access approach for surgical management of cardiac tumors. AB - BACKGROUND: Following our experience with minimally invasive valve replacement operation, we utilized this technique for surgical management of cardiac tumors. METHODS: Between April 1997 and September 1999, 5 consecutive patients with cardiac tumors underwent minimally invasive excision of the tumors. The patients were 4 women and 1 man with an age range of 32 to 50 years. The tumor was located in the left atrium in 4 patients and the right atrium in 1 patient. The common presenting symptoms were dyspnea on exertion (100%), chest pain (60%), palpitation (60%), and transient ischemic attack (20%). Diagnosis was established preoperatively by echocardiography only. RESULTS: In 2 patients the approach was right parasternal and the subsequent 3 patients had direct-access partial sternotomy. The myxoma was resected transseptally in all patients. There was no hospital mortality. One patient had postoperative embolic episode leading to left hemiparesis. Follow-up did not reveal any complication related to this technique and all were in New York Heart Association (NYHA) functional class I. CONCLUSIONS: Minimal access partial sternotomy is an effective approach that adheres to all the identified surgical principles in successful removal of these tumors. The smaller incision does not compromise the efficacy or safety of the operation, reduces hospital stay, and has a good cosmetic result. PMID- 11016381 TI - Combined stentless mitral valve implantation and radiofrequency ablation. AB - BACKGROUND: The aim of this study was to evaluate the results of combined stentless mitral valve (SMV) replacement and intraoperative radiofrequency ablation for chronic atrial fibrillation (IRAAF) to restore physiologic hemodynamic function. METHODS: Since July 1998 12 patients (72+/-4 years, 10 women, mitral stenosis/mitral incompetence 8/4, NYHA 3.3+/-0.4, CI 1.8+/-0.5) had SMV implantation and received additional IRAAF by inducing continuous left atrial lesion lines from the MV annulus to all four pulmonary veins and to the atriotomy. RESULTS: The flexible SMV was implanted at the papillary muscles and at the annulus using a conventional (n = 6) or a minimally invasive approach (n = 6). Sinus rhythm was successfully restored in 10 of 12 patients with 6- and 12 months' follow-up; 2 required DDD-pacemaker implantation. However, in the early postoperative period several interventions including medical treatment (sotalol or amiodarone) in 9 and electrical cardioversion in 7 patients was required. Two patients required reinterventions: 1 cardioversion and 1 amiodarone medication after 3 and 6 months, respectively. At echocardiography the SMV demonstrated good hemodynamic function and atrial contraction. CONCLUSIONS: Restoration of physiologic cardiac function by SMV implantation and IRAAF is advantageous and no further anticoagulation is required. PMID- 11016382 TI - Right heart circulatory support facilities coronary artery bypass without cardiopulmonary bypass. AB - BACKGROUND: Revascularization of the posterior wall often causes hemodynamic instability in beating heart coronary artery bypass (CAB). Our previous clinical studies have shown that tilting the heart primarily alters right-heart hemodynamics. The purpose of this study was to evaluate right-heart support in clinical cases. METHODS: Seventeen patients underwent beating heart CAB with right-heart support. The right-heart support system (A-Med Systems, West Sacramento, CA) consisted of a coaxial cannula placed through the right atrium and the tip of the cannula positioned in main pulmonary artery. Blood was removed from the right atrium and returned to the main pulmonary artery. RESULTS: Elective beating heart CAB was accomplished successfully in 17 patients with right-heart support. Anastomoses performed were left anterior descending coronary artery (11), diagonal (3), circumflex (5), obtuse marginal artery (11), and right coronary artery (10). Right-heart support between 1 and 3 L/min improved hemodynamics especially in the circumflex position. No device-related patient incidents occurred, nor were there incidents of infection or air embolism. All 17 patients were discharged to their homes. CONCLUSIONS: The right-heart support system was safe without complications. Exposure of the posterior wall was possible in all cases without hemodynamic compromise. PMID- 11016383 TI - Endoscopic saphenous vein harvesting: initial experience and learning curve. AB - BACKGROUND: Saphenous vein remains an elective conduit for up to 85% of coronary bypass operations. It is obtained through one or numerous skin incisions, with a reported morbidity varying from 5% to 25%. The endoscopic vein harvesting (EVH) technique was developed to minimize this morbidity and to improve clinical outcomes. The aim of this study was to review the feasibility of this method, its learning curve, and changing results in a group without previous experience in this procedure. METHODS: Between July 1998 and October 1999, 179 patients for coronary artery bypass grafting underwent EVH (Vasoview Guidant, USA "double access" and Uniport), by two operators. Results were reported based on time of harvesting, length of conduits, technical details, and clinical outcomes, and divided into six groups of 30 consecutive patients each. RESULTS: Patient demographics were as follows: 86.03% were male, aged 64.3+/-9.12 years (range, 43 to 92 years), with diabetes mellitus in 28.49%, obesity in 18.43%, and vascular disease in 11.17%. The EVH method was limited to the thigh in 77.65% of cases and extended to the leg in 22.35%. Patients received an average of 2.45+/-0.58 incisions and obtained conduits had a mean length of 34.96+/-9.65 cm (range, 15 to 70 cm). The number of venous bypasses per patient was 1.30+/-0.59. Mean time of EVH was 47.24+/-19.84 minutes (range, 15 to 120), with a length-time index of 0.85+/-0.36. Primary success was achieved in 95.54%, with crossover to open technique in 4.46%. General morbidity was 8.9%, with hematoma in 1.11%, skin necrosis in 1.11%, infection in 6.7%, and readmission in 1.11%. CONCLUSIONS: Endoscopic vein harvesting is a feasible and reproducible method, with a typical learning curve, acceptable morbidity, and unquestionable benefits for coronary artery bypass graft patients. PMID- 11016384 TI - Use of three-dimensional computed tomography images in deciding the approach for ministernotomy operations. AB - BACKGROUND: In ministernotomy operations for valvular and congenital heart diseases, several approaches exist. Because topographical relationships of the sternum, ribs, heart, and great vessels differ slightly with each patient, three dimensional (3D) computed tomography (CT) images are valuable in deciding the best approach for each patient. METHODS: Ministernotomy patients were studied preoperatively using 3D-CT images. By subtracting some parts of the sternum and the ribs from the 3D images, the topographical relationships of the structures were revealed. RESULTS: Different methods of approach (ie, the site, shape, and length of partial sternotomy) were compared on the 3D images of each patient to decide the most appropriate incision. The images were shown to resemble closely the actual operative field. CONCLUSIONS: Three-dimensional CT images are valuable in preoperative planning of ministernotomy, and are essential to individualized planning for each operation. PMID- 11016385 TI - Cerebral microemboli during minimally invasive and conventional mitral valve operations. AB - BACKGROUND: Intraoperative cerebral microembolism may cause postoperative neurologic damage. The aim of this study was to determine the frequency of cerebral microembolic signals (MES) during minimally invasive surgery (MIS) and conventional (conv.) mitral valve operations and to determine the association of MES with various stages of the operation. METHODS: Intraoperative computer-aided transcranial Doppler measurements were performed to detect cerebral microemboli in 21 patients undergoing MIS and in 14 patients undergoing conv. mitral valve operation. We calculated the mean embolic rate for three time periods: P1, start of the operation until aortic clamping; P2, aortic clamping until clamp removal; and P3, declamping until end of surgery. RESULTS: There was no significant difference in the total number of detected cerebral MES between both patient groups (MIS 1,014+/-753, conv. 937+/-519; NS). In both groups, the highest number of MES were detected during the third time period when the heart regained effective ejection (MIS 875+/-746, conv. 680+/-462; p > 0.5). CONCLUSIONS: Transcranial Doppler was useful to detect cerebral microemboli in MIS and conv. mitral valve operation. We found no increased risk of cerebral microembolism during the minimally invasive method compared with the conventional technique. PMID- 11016386 TI - Hemodynamic effects of carbon dioxide insufflation during endoscopic vein harvesting. AB - BACKGROUND: A prospective study was performed assessing the hemodynamic effects of carbon dioxide (CO2) insufflation during endoscopic vein harvesting (EVH) using the Guidant Vasoview Uniport system. METHODS: Five hemodynamic and respiratory parameters (end-tidal carbon dioxide, arterial partial pressure of carbon dioxide, mean arterial pressure, mean pulmonary arterial pressure, and cardiac output), were measured in 100 consecutive patients undergoing EVH with CO2 insufflation. Data were obtained prior to commencement of EVH, 15 minutes after commencement, and 5 minutes after completion of the vein harvesting. RESULTS: No adverse hemodynamic effects were observed during CO2 insufflation. Specifically, average mean arterial pressure went from 88.77+/-9.64 to 89.13+/ 8.60 to 88.24+/-8.71 mm Hg before, during, and after endoscopic vein harvesting (p = 0.291). Likewise, average mean pulmonary artery pressures were 19.76+/-4.75, 20.05+/-4.48, and 20.05+/-4.62 mm Hg (p = 0.547); and average cardiac output was 4.25+/-0.74, 4.22+/-0.73, and 4.23+/-0.69 L/min (p = 0.109) at those three intervals. Additionally, there was no evidence of significant systemic absorption of CO2 as reflected in average arterial PCO2, which remained steady at 37.42+/ 5.19, 37.51+/-4.59, and 38.10+/-4.80 mm Hg (p = 0.217); and average end-tidal CO2, which was 32.10+/-3.66, 32.50+/-3.47, and 32.38+/-3.33 mm Hg (p = 0.335). In a subset of 20 patients with elevated pulmonary arterial pressure (more than 32 mm Hg), there was also no significant change in any of the parameters. CONCLUSIONS: Carbon dioxide insufflation during EVH leads to no adverse hemodynamic consequences or systemic CO2 absorption. The technique appears to be safe and well tolerated. PMID- 11016387 TI - Pain is significantly reduced by cryoablation therapy in patients with lateral minithoracotomy. AB - BACKGROUND: In minimally invasive cardiac surgery use of a lateral minithoracotomy is associated with early postoperative rehabilitation but also relatively high pain levels during the first 3 postoperative days. Cryoablation therapy was evaluated prospectively. METHODS: From April 1999 until September 1999, 57 patients underwent lateral minithoracotomy for mitral valve operation (n = 18) or minimally invasive direct coronary artery bypass grafting (n = 39). Intraoperatively, patients were randomly assigned to cryoablation or intercostal application of local anesthetic agents. A standardized questionnaire was used for prospective pain assessment on postoperative days 1 to 7. RESULTS: From postoperative day 1 to 7 pain levels declined in all groups. Overall pain levels were significantly lower in the cryo group than in the control group (p < 0.0001, GLM). According to diagnoses, pain levels were significantly lower after MIDCABG and cryo versus control; after mitral valve operation they were lower in the cryo group and almost reached significance. CONCLUSIONS: Cryoablation is easy to perform and leads to a significant reduction in pain and lower request for additional pain medication after lateral minithoracotomy in minimally invasive cardiac operation. PMID- 11016388 TI - Wrist-enhanced instrumentation: moving toward totally endoscopic coronary artery bypass grafting. AB - BACKGROUND: With the introduction of the da Vinci robotic surgical system (Intuitive Surgical, Mountain View, CA) into minimally invasive cardiac operations the outlook for performing coronary artery bypass operations "closed chest" became a reality. METHODS: Between May 1999 and December 1999 this new wrist-enhanced instrumentation was used in 61 patients. Six patients suffering from single-vessel coronary artery disease and one female patient with double vessel disease underwent totally endoscopic coronary artery bypass. Thirty-seven patients with single-vessel disease underwent a minimally invasive direct coronary artery bypass procedure. Seventeen patients with double-vessel disease were treated using the robotic-enhanced Dresden technique. RESULTS: Perioperative survival was 100%. In all patients the internal mammary arteries were safely harvested endoscopically and had excellent quality. In both totally endoscopic coronary artery bypass groups all patients were operated on through three stab incisions. CONCLUSIONS: Our preliminary experience with this new surgical technique using robotic-enhanced minimally invasive methods for coronary artery disease promotes optimism regarding further development of these procedures and application in patients with coronary artery disease. PMID- 11016389 TI - A method for mediastinal drainage after cardiac procedures using small silastic drains. AB - BACKGROUND: It has been standard teaching in cardiac surgery that drainage of the mediastinum following cardiac surgical procedures is best accomplished using rigid large-bore chest tubes. Recent trends in cardiac surgery have suggested less invasive approaches to a variety of diseases. Difficult drainage problems in the field of general surgery including hepatic and pancreatic collections have been drained successfully with smaller flexible drains for many years. Additionally, many difficult to reach collections in the chest have been drained by invasive radiologists using small pigtail catheters. METHODS: We have introduced drainage of the mediastinum using 10-mm flexible, flat, fluted Blake drains. To date, we have used these drains in more than 100 cardiac operations including coronary artery bypass grafting, valve repair/replacements, combined coronary artery bypass grafting/valve operations, heart transplants, septal defects, and mediastinal tumors. RESULTS: We have demonstrated that this form of drainage is as good as using large-bore chest tubes with no significant risk of bleeding or tamponade. Additionally, use of these tubes is less painful, allows more mobility, and earlier discharge with functioning drains in place if necessary. CONCLUSIONS: Larger chest tubes are not necessarily better when it comes to draining the mediastinum. The actual area of ingress through the sideholes is considerably less than the surface area provided by the fluted Blake drain. We believe that this system can replace standard chest tubes. PMID- 11016390 TI - Effects on coronary endothelial function of the Cohn stabilizer for beating heart bypass operations. AB - BACKGROUND: The effects of different stabilizing methods for minimally invasive beating heart coronary artery bypass grafts (CABG) on coronary endothelial function are unknown. METHODS: We compared the effects on endothelial function of the Cohn stabilizer (used with proximal snaring by Retract-o-tape silicone air cushion) and a coronary shunt (CTS flow coil shunt) on an in vivo model of beating heart CABG. The two techniques were applied for 15 minutes on porcine epicardial coronary arteries. Control rings were taken from the same coronary artery. Endothelial function of control and instrumented arterial rings was studied in organ chamber experiments. Evaluation of endothelial coverage was performed with silver nitrate staining. RESULTS: Endothelium-dependent relaxation to serotonin and bradykinin was significantly decreased in the shunt group compared to control, Cohn stabilizer, and snare groups. There were no significant differences in the endothelium-independent relaxation to sodium nitroprusside between groups. CONCLUSIONS: Greater endothelium-dependent relaxation with the Cohn stabilizer suggests better preservation of endothelial coverage at the site of application and reduced propensity for coronary spasm and later development of intimal hyperplasia. PMID- 11016391 TI - Instrumental validation of percutaneous transmyocardial revascularization: follow up data at one year. AB - BACKGROUND: Despite the clinical efficacy of percutaneous transmyocardial revascularization (PTMR), up to date there are still no instrumental validations to demonstrate both the improved perfusion of treated areas and cardiac function. METHODS: During the first year of follow-up after PTMR, 27 patients (group A) underwent 99mTc MIBI exercise-single photon emission tomography (SPET), while 30 patients (group B) underwent serial transthoracic echocardiography (TTE) evaluations with analysis of cardiac volumes and subendocardial layer thickness in systole. RESULTS: All 57 patients had a significant angina Canadian Cardiovascular Society (CCS) class improvement. Group A patients (75%) had improved exercise-SPET perfusion in treated areas at 12 weeks after PTMR, and at the next follow-up. Group B patients had non-significant reduction in global volume and no significant change in ejection fraction. However, there was an improvement in thickness of the subendocardial-treated areas in systole that persisted during follow-up. CONCLUSIONS: The use of SPET and TTE validates the clinical efficacy of PTMR. PMID- 11016392 TI - Capture of particulate emboli during cardiac procedures in which aortic cross clamp is used. International Council of Emboli Management Study Group. AB - BACKGROUND: Previous studies have shown that atheroemboli are associated with neurologic complications following cardiac operation. Additionally, it has been demonstrated that embolization is closely related to the application and removal of ascending aortic cross-clamps. METHODS: A prospective registry of 304 patients was initiated to describe patient selection, procedures, particulate capture, and clinical outcomes after intraaortic filtration in patients undergoing cardiac surgical procedures with cardiopulmonary bypass and median sternotomy. Prior to the removal of the cross-clamp, the intraaortic filter (EMBOL-X Inc, Mountain View, CA) was deployed and left in place until the patient was weaned from extracorporeal circulation. Upon removal, filters were fixed in formalin and shipped to a core laboratory for examination. RESULTS: Mean patient age was 68 years (range 25 to 88 years), 40% had ascending aortic calcification, 59% of the procedures were coronary artery bypass grafting (CABG), 20% were valve replacement or repair, and 12% were a combination of CABG and valve surgery. Aortic cross-clamps were used in nearly all cases (302 of 304 patients); partial clamps were used in 84% of the CABG procedures. To date, 243 filters have been examined. Sixty-two percent of the filters analyzed revealed fibrous atheroma, 2% grumous atheroma, and 6% epiaortic debris. Platelet and fibrin strands were found in 52% of the filters and 22% contained evidence of thrombus or red blood cells. CONCLUSIONS: These findings from the International Council of Emboli Management Registry confirm that particulates are released during cardiac surgical procedures using the aortic cross-clamp. Continued observational and randomized studies are necessary to confirm the clinical relevance of particulate extraction. PMID- 11016393 TI - Ventricular containment as an adjunctive procedure in ischemic caridomyopathy: early results. AB - BACKGROUND: Ventricular containment with custom-made polyester mesh is an evolving technique that has been studied in experimental animals with heart failure with good results. METHODS: Five patients with symptomatic heart failure and ischemic cardiomyopathy were enrolled in a Phase I study, and underwent ventricular containment with custom-made polyester mesh along with coronary artery bypass grafting. Four patients had additional ventricular reconstruction of large myocardial scars. RESULTS: All patients were in NYHA functional class III at the time of their operation with a mean ejection fraction of 27.4%+/-6.6%. There were no deaths. Mean postoperative ejection fraction was 35.1%+/-12.6% (p = 0.16). Left ventricular end-diastolic diameter fell from 63.2+/-1.6 mm preoperatively to 50.6+/-5 mm, postoperatively (p = 0.004). There was no evidence of diastolic dysfunction or pericardial constriction on intra- or postoperative echocardiography. At a mean follow-up of 180 days all patients were in NYHA class I with no readmissions for heart failure. Repeat coronary angiography at 6 months revealed patent grafts in all patients. CONCLUSIONS: Ventricular containment with a customized mesh may be performed safely as an adjunct to conventional cardiac operation in patients with symptomatic heart failure. Longer follow-up with an expansion of the study will help delineate the long-term effects of this therapy. PMID- 11016394 TI - Outcome of geometric endoventricular repair in impaired left ventricular function. AB - BACKGROUND: Traditionally, repair of left ventricular aneurysms has been limited to patients with large localized ventricular aneurysms. Repair of dyskinetic segments in the setting of poor left ventricular function is still contentious. METHODS: Forty patients underwent geometric endoventricular repair, a new technique of ventricular aneurysm repair, over a 2-year period. Two groups of patients undergoing coronary artery bypass grafting (CABG) for left ventricular dysfunction in the same time period were reviewed. Group 1 comprised 23 consecutive patients who underwent geometric endo-ventricular repair along with CABGs, whereas group II consisted of 22 patients who underwent CABG alone. RESULTS: The early mortality was 9.1% in group I (1 cardiac, 1 noncardiac) and 0 in group II (NS). New York Heart Association class was remarkably improved from 3.4 to 1.4 (p < 0.05) in group I and to a lesser extent in group II (3.7+/-0.5 versus 2.3+/-0.5). Diastolic dimension of left ventricle was significantly reduced from 5.6 cm to 4.4 cm (p < 0.05) in group I and virtually unchanged in group II. There was one late death in each of the groups. CONCLUSIONS: This technique of geometric left ventricular aneurysm repair is useful in patients with dyskinetic segments and may help in reducing cardiac size. PMID- 11016395 TI - Symptomatic improvement after transmyocardial laser revascularization: how long does it last? AB - BACKGROUND: The aim of this study was to determine whether short-term clinical improvement after isolated transmyocardial holmium laser revascularization (TMLR) in patients with coronary artery disease not amenable to traditional treatment is maintained through a longer follow-up. METHODS: Between November 1995 and June 1999 34 patients underwent TMLR (mean age, 67+/-7 years); previous revascularization procedures had been performed in 76%. Preoperatively, mean angina class was 3.6+/-0.5 in 12 patients with unstable angina; mean left ventricular ejection fraction was 47%+/-9%. RESULTS: There was 1 early death due to low cardiac output. Mean duration of TMLR and of the entire operation was 25+/ 12 minutes and 125+/-43 minutes, respectively. There were no major postoperative complications; mean hospital stay was 8+/-4 days. There were 8 late deaths caused by stroke (2 patients), cardiac failure (1 patient), and myocardial infarction (5 patients). Follow-up of current survivors ranges from 4 to 48 months (mean, 32+/ 12 months). At 1-year follow-up mean angina class was 1.8+/-0.8; but at a later follow-up (mean, 35+/-10 months) it significantly increased to 2.2+/-0.7 (p = 0.005). Three-year actuarial survival was 76%+/-8% and freedom from cardiac events 44%+/-10%. CONCLUSIONS: Our results show that after initial clinical improvement many patients experience return of angina or cardiac events; this questions the long-term symptomatic benefit of TMLR. PMID- 11016396 TI - Inflammatory response and angiogenesis after percutaneous transmyocardial laser revascularization. AB - BACKGROUND: The aim of our study was to investigate the inflammatory response immediately after percutaneous transmyocardial laser revascularization (PTMR) along with the underlying mechanism of angiogenesis. METHODS: Patients with angina pectoris underwent coronary angiography and were divided into two groups. Group A (n = 10) included patients with obstructed vessels who received PTMR, whereas group B (n = 5) comprised patients who had normal coronary arteries. Blood levels of neutrophils, procalcitonin, troponin-I, myoglobin, and creatine kinase (CK) mass were evaluated in each patient before angiography and monitored up to 48 hours after the procedure. Six patients were injected with 99mTc leukoscan approximately 60 to 90 minutes after PTMR. During the 240 to 300 minutes after the radionuclide administration, single photon emission tomography (SPET) was performed and compared with conventional 99mTc-sestamibi-SPET. RESULTS: A significant increase in blood levels of neutrophils and procalcitonin was observed in group A only (p < 0.005). A slight but significant increase of troponin-I was evident in the same group (p < 0.05), and a distinct myocardial uptake of 99mTc-Leukoscan-SPET was observed in each patient along homologous regions treated by PTMR. CONCLUSIONS: The increased amount of neutrophils (both circulating and inside the treated myocardial areas) along with the raised levels of procalcitonin were the immediate reactions to PTMR. This systemic and intramyocardial inflammatory response is the underlying mechanism that gives rise to angiogenesis. PMID- 11016398 TI - A biomechanical and spectroscopic study of bone from rats with selenium deficiency and toxicity. AB - Selenium, being an essential mineral in the mammalian diet, is important in providing protection against oxidative damage. Numerous in vitro studies of selenium compounds reveal a very high correlation between catalytic activity of selenium compounds and toxicity. The present study was designed to investigate the effects of dietary selenium on the biomechanical properties of bone. New born rats of both sexes were fed with either a control, or a selenium- and vitamin E deficient, or a selenium-excess and vitamin E-adequate diet. We obtained the stiffness (modulus of elasticity) of bones (femur and tibia) by tensile test for all groups considered. Both the deficient and the excess groups have decreased biomechanical strength with respect to the control group. To support our biomechanical results for both experimental groups, X-ray diffraction analysis and FTIR spectroscopic study were performed on the femurs and tibiae. The X-ray diffraction analysis showed that the intensities of the peak observed at around 2theta degrees = 31.820, in the control femur and tibia are stronger than the intensities of the corresponding peak of two experimental groups. In FTIR spectroscopy, the disappearance and/or reduction of the intensities of some carbonate bands in the two experimental groups indicate that there is a decrease in crystallinity and mineral contents which, together with X-ray diffraction analysis, correlate very well with the biomechanical data. PMID- 11016397 TI - Differences in the growth inhibition of cultured K-562 cells by selenium, mercury or cadmium in two tissue culture media (RPMI-1640, Ham's F-10). AB - Effects of some metals on the growth of cultured human erythroleukemia K-562 cells were investigated when grown in two different types of media based upon RPMI-1640 or Ham's F-10. The study on proliferation, using RPMI-1640 supplemented with sodium selenite, selenomethionine, mercuric chloride, methylmercuric chloride and cadmium nitrate showed no inhibition of growth at concentrations of 2.5, 25, 25, 2.5 and 25 microM, while at 75, 250, 50, 5 and 50 microM toxicity was apparent. Selenite at 5-50 microM and selenomethionine at 50-100 microM inhibited the growth. In Ham's F-10 supplemented with the same compounds no inhibition was found at concentrations of 5, 10, 25, 1 and 50 microM, while at 50, 100, 50, 5 and 75 microM toxic effects were noted. Selenite 10 microM and selenomethionine 25-50 microM inhibited the proliferation. Measurements of trace element levels in pellets of K-562 cells grown in RPMI-1640 or Ham's F-10 unveiled higher cell contents of cadmium and selenium in cells grown in RPMI 1640, being consistent with higher concentrations of these elements in that medium. Manganese and mercury concentrations were higher in cells grown in Ham's F-10 correlating with a higher medium concentration of these elements. The growth responses and cellular uptake differed between the metals and the selenocompounds and although extrapolating the results to humans is difficult the selenium exposures were in approximately the same order of magnitude as in human exposures. The compounds could be ranked according to decreasing toxicity as: methylmercuric chloride > mercuric chloride, cadmium nitrate, sodium selenite > selenomethionine. PMID- 11016399 TI - Influence of aluminium on the immune system--an experimental study on volunteers. AB - The purpose of this study was to examine whether oral exposure to aluminum (Al) can affect the human immune system. Eighteen healthy volunteers (mean age 42, 28 57 yr) were divided into a test group (9 females, 4 males) and a referent group (3 females, 2 males). Over 6 weeks, the test subjects ingested 10 ml of antacid (aluminum hydroxide, 59 mg Al/ml) three times daily. Aluminum was analyzed in urine before and during the exposure period (ICP-MS). Blood samples were used for analysis of lymphocyte subpopulations, mitogen-induced lymphocyte proliferation and in vitro production and circulating plasma concentrations of immunoglobulin (Ig) A, IgG, IgM, interleukin (IL) -2 and IL-4. Urinary Al concentration in the test subjects was approximately 10- to 20-fold higher than in the referent group during exposure. This indicates that ingestion of an Al-containing antacid is associated with an Al absorption far above that originating from food and drinking water. In both referents and test subjects the lymphocyte subpopulations, lymphocyte proliferation and the in vitro Ig and IL production showed similar, time-dependent changes before as well as during the exposure period. No major differences were seen between the referent and test groups regarding the immune parameters, except for a slightly smaller CD8+CD45R0+ population (primed cytotoxic T-cells), in the exposed individuals as compared to the referents. The results also show that subjects on antacid therapy may constitute a suitable population for studying biological effects of high-dose oral exposure to Al. PMID- 11016400 TI - DNA sequence analysis of the tellurite-resistance determinant from clinical strain of Escherichia coli and identification of essential genes. AB - The tellurite-resistant Escherichia coli strain KL53 was found during testing of the group of clinical isolates for antibiotics and heavy metal ion resistance (Burian et al. 1990). Determinant of the tellurite resistance of the strain was located on the large conjugative plasmid pTE53 and cloned into pACYC184. Three different Ter clones harboring pLK2, pLK18 and pLK20 were isolated (Burian et al. 1998). The smallest functional Ter clone harboring pLK18 was chosen for further analysis. Plasmid pLK18 have been subcloned to obtain convenient DNA fragments for sequencing of tellurite-resistance determinant. Sequencing of this DNA fragments provided complete DNA sequence of the determinant, 5,250 bp in size. The sequence has been compared with nucleotide and protein databank (BLAST programs) and significant homology with the three known operons coding for tellurite resistance has been found (determinat on plasmid pR478 from Serratia marcescens, on plasmid pMER610 from Alcaligenes sp. and chromosomal tellurite resistance genes from Proteus mirabilis). We identified 5 ORFs coding for 5 genes named terB to terF. The clone harboring pLK18 was subjected to the transposition with Tn1737Km to disrupt determinant of the tellurite resistance. Plasmid DNA of several clones containing pLK18 with Tn1737Km was isolated to locate the target site of Tn1737Km. Analyses showed, the genes terB, terC, terD and terE are essential for conservation of the resistance whereas the gene terF is not important in this respect. PMID- 11016401 TI - Zinc kinetics in insulin-dependent diabetes mellitus patients. AB - Zinc has an important role in the control of carbohydrate metabolism, and diabetic patients are at risk for zinc deficiency. However, there are conflicting data concerning nutritional zinc status. In order to investigate this topic, 10 normal and 10 insulin-dependent diabetic patients were studied following venous zinc tolerance test. Our results found no evidence of zinc deficiency or of changes on the kinetic parameters of zinc in patients with insulin-dependent diabetes mellitus following a venous zinc tolerance test. PMID- 11016402 TI - A pyoverdine from Pseudomonas putida CFML 90-51 with a Lys epsilon-amino link in the peptide chain. AB - From Pseudomonas putida CFML 90-51--a hospital isolate--a pyoverdine was obtained which is characterized by the unusual linkage by the epsilon--rather than the alpha-amino group of Lys in the peptide chain. The structure elucidation by spectroscopic methods and degradation reactions is reported. PMID- 11016403 TI - Copper-release from yeast Cu(I)-metallothionein by nitric oxide (NO). AB - The reaction of yeast Cu-MT with nitric oxide (NO) was examined. A release of copper from the Cu(I)-thiolate clusters of the protein by this remarkably important reagent was observed in vitro. The characteristic spectroscopic signals of the Cu(I)-thiolate chromophores levelled off in the presence of a two-fold molar excess of NO expressed per equivalent of thionein-copper as monitored by UV electronic absorption, circular dichroism and luminescence emission. At the same time all of the copper became EPR detectable. The oxidized metal ions could easily be removed from the protein moiety by gelfiltration. The reversibility of the copper releasing process is of special interest. The specific fluorescence and dichroic properties of the previously demetallized protein could be recovered up to 85% under reductive conditions. Moreover, no difference in the electrophoretic behaviour was seen compared to the untreated Cu-MT. Thus, NO may act as a potent metabolic source for the transient copper release from Cu-MT. In the course of an oxidative burst this highly Fenton active copper is able to improve the efficacy of biological defence mechanisms. PMID- 11016404 TI - Changes in lipid peroxidation, the redox system and ATPase activities in plasma membranes of rice seedling roots caused by lanthanum chloride. AB - Highly purified plasma membranes were isolated by aqueous two-phase partitioning from rice (Oryza sativa) seedling roots. The effects of lanthanum chloride (LaCl3) on the activities of lipid peroxidation, the redox system and H+-ATPase, Ca2+-ATPase of plasma membranes were studied. The lipid peroxidation of plasma membranes could be depressed by certain low concentrations of LaCl3 and enhanced by high concentrations of LaCl3, while the lipid peroxidation was also dependent on the plasma membrane protein and incubation time. The relative activity of O2 uptake of plasma membranes was inhibited by all tested LaCl3 concentrations. In contrast, the reduction rate of Fe(CN)6(3-) by plasma membranes was stimulated below 40 microM of LaCl3, but was reduced above 60 microM of LaCl3. The relative activities of both H+-ATPase and Ca2+-ATPase increased constantly from control to LaCl3 of concentration 60 microM where the activities of both enzymes were the maximum. but decreased remarkably at 80 microM LaCl3 concentrations various LaCl3 were added to culture solutions. In the other measurement case in which various LaCl3 concentrations were added directly to reaction medium and the plasma membrane vesicles only came from the control cultured rice seedling roots, the response of H+-ATPase activity to La3+ was similar to the response in culture solution. However, the La3+ concentration was only 20 microM when the activity of H+-ATPase was the maximum. In contrast to the case of LaCl3 addition to culture solution, Ca2+-ATPase activity was inhibited by all concentrations of La3+ which were added directly to the reaction medium. The above results revealed that REEs inhibited electron transfer from NADH to oxygen in plant plasma membranes, depressed the production of active oxygen radicals, and reduced the formation of lipid peroxides through plasma membrane lipid peroxidation. REEs ions also enhanced the H+ extrusion by both standard redox system and H+-ATPase in plasma membranes at certain concentrations. A possible role for the plant cell wall in REEs effects on plasma membranes was also suggested. PMID- 11016405 TI - Salicylic acid is not a bacterial siderophore: a theoretical study. AB - Using a newly available program for calculating the concentrations and speciation of various ions (Pettit, LD & Powell KJ, 'SolEq' Academic Software, 1999), we have calculated that at pH 7 the amount of free Fe(III) present in an aqueous solution is 1.4 x 10(-9) M and not 10(-18) M as is usually quoted. In the presence of salicylic acid, included in the calculations at 10(-4) M, the solubility of Fe(III) is increased to only 9.8 x 10(-9) M suggesting that salicylate is unable to act as a siderophore although it is produced as an extracellular product by several bacterial genera when grown iron deficiently. In the presence of 40 mM phosphate, the soluble Fe(III) concentration is decreased by 10(4) at pH 7 and again this is hardly affected by the presence of salicylate. Thus, for microorganisms grown either in vitro or in vivo, salicylate is unlikely to function as a iron solubilizing agent. The same conclusions may also apply to 2,3-dihydroxybenzoic acid. PMID- 11016406 TI - Voltage-clamp: a useful approach to study in vitro duodenal iron transport in the mouse. AB - The mechanism(s) controlling iron absorption remain(s) uncertain despite the progress in the identification of genes selectively expressed in the duodenum. The availability of experimental models of iron absorption is critical to the clarification of such mechanism(s). In the present study, a simple method for studying in vitro iron absorption in mouse duodenum is described. Short circuit current, open circuit potentials and epithelial conductances were measured in mouse duodenal segments. Also, unidirectional 55Fe fluxes at different pH conditions were measured in mice with varying iron status. The findings reinforce evidence for an adaptive response of the iron absorptive process according to the body iron status. Significant differences are demonstrated between iron fluxes measured in normal and parenterally iron loaded mice and at acidic compared to neutral pH environment. Also, a significant difference was observed between transepithelial potential measured in duodenum from iron-loaded compared to untreated mice. Advances in the understanding of the mechanism(s) of iron absorption can be brought by the application of voltage-clamp techniques to the electrophysiologic study of iron overload mouse models. PMID- 11016407 TI - Copper uptake and intracellular distribution in the human intestinal Caco-2 cell line. AB - The apical uptake of 64CuCl2 was investigated in human differentiated intestinal Caco-2 cells grown on permeable supports. At pH 6.0 in the apical compartment, the uptake of copper was linear over the first 6 min and between 10 and 80 microM CuCl2 exhibited non-saturable transport kinetics. In addition, copper uptake was energy-independent, affected by the valency state of copper, preferring Cu(II) over Cu(I), and not influenced by high (10 mM) extracellular calcium. The intracellular distribution of copper was investigated by FPLC at different times of uptake ('pulse') and of 'chase'. Intracellular copper initially bound predominantly to low molecular weight components (i.e., glutathione). and subsequently shifted to higher molecular weight components such as metallothionein and Cu,Zn superoxide dismutase. PMID- 11016408 TI - Impact of lead exposure on pituitary-thyroid axis in humans. AB - Thyroid function tests (serum levels of thyroxine-T4, triiodothyronine-T3 and thyroid stimulating hormone-TSH) were performed in fifty-eight men (mean age: 31.7 +/- 10.6 years; mean duration of lead exposure: 156.9 +/- 122.7 months). These subjects were exposed to lead either as petrol pump workers or automobile mechanics. The mean whole blood lead (Pb-B) levels were 2.49 +/- 0.45 micromole/l (51.90 +/- 9.40 microg/dl) in the lead exposed workers and were approximately 5 times higher than in the control (n = 35) subjects. No significant alteration was seen in their mean T3 and T4 levels as compared with the controls. Interestingly, T3 was significantly lower with the longer (210 months) exposure time in comparison with the group having shorter (29 months) exposure duration. The mean TSH levels were significantly (p < 0.01) higher in workers exposed in comparison with the control group. This rise in TSH was independent of exposure time, but it was definitely associated with the Pb-B levels. The increase being more pronounced with mean Pb-B levels of 2.66 +/- 0.2 micromole/l (55.4 +/- 4.25 microg/dl) when compared with the group having mean levels of 1.51 +/- 0.30 micromole/l (31.5 +/- 6.20 microg/dl). The rise is TSH associated with Pb-B levels was only statistical valid, however, the levels fall within the normal laboratory range. We thus conclude that the Pb-B levels of > or = 2.4 micromole/l (50 microg/dl) could enhance the pituitary release of TSH without having any significant alterations in the circulating levels of T3 and T4. PMID- 11016409 TI - Back to fundamentals: anatomy-based physiological bioengineering. PMID- 11016410 TI - Quasiperiodic circus movement in a loop model of cardiac tissue: multistability and low dimensional equivalence. AB - This paper discusses the dynamics of the circus movement around a one-dimensional continuous and uniform loop of model cardiac cells. The membrane ionic currents are represented by a modified Beeler-Reuter formulation. The description of the quasiperiodic regimes initiated in a previous paper is completed and their equivalence with those predicted by an integral-delay model is discussed. The two models differ in the number of quasiperiodic modes of reentry that can be sustained and in the type of bifurcation by which they appear. These differences are the consequences of the modulation of the repolarization phase by diffusion in the ionic model. A modified version of the integral-delay model that includes a spatial averaging in the calculation of the action potential duration is shown to suppress the discrepancies. PMID- 11016411 TI - Effects of material properties and geometry on electrocardiographic forward simulations. AB - This paper investigates the effect on torso potentials of changes in the material properties of the torso volume conductor and changes in the relative geometry of the heart and torso. The investigations are performed using a number of forward simulations with a high-order coupled finite element/boundary element torso model. This torso model contains descriptions of the epicardium, lungs, skeletal muscle (with a continuously varying fiber field) and subcutaneous fat. The number of nodes, elements and solution degrees-of-freedom used in the computational torso model are considerably smaller than previous torso models of similar complexity. The successful forward simulations in this paper hence demonstrate the use of the high-order coupled approach with realistic problems. The results of the torso simulations show that the torso inhomogeneities do affect the torso potentials but do not affect the distribution or pattern of the torso potentials. The inhomogeneities considered are found to have a varying, but important, effect on the torso potentials. The effect of the subcutaneous fat is found to be more important and the effect of the skeletal muscle is found to be less important than previous reports in the literature. The results also show that the relative geometry of the heart and torso is very important in determining the torso potential magnitudes and distributions. PMID- 11016412 TI - The role of the crista terminalis in atrial flutter and fibrillation: a computer modeling study. AB - Although atrial fibrillation is a common arrhythmia, the underlying mechanisms are incompletely understood. Recent studies have determined the role of the crista terminalis in the mechanisms of a simpler arrhythmia, atrial flutter. We hypothesize that as transverse coupling across the crista terminalis increases, the activation pattern that results is less like typical atrial flutter and more like atrial fibrillation. 6480 Van Capelle elements were coupled in an icosahedron, simulating the right atrium. Atrial simulations were created which incorporated no heterogeneity, heterogeneous coupling, heterogeneous effective refractory periods, and both heterogeneous coupling and effective refractory periods. When the entire crista terminalis was uncoupled, typical atrial flutter occurred. When transverse coupling allowed activation to propagate across the crista terminalis, the flutter cycle length decreased (p<0.0001). In addition, when heterogeneity was present, both the coefficient of variation of cycle length and the number of activation wavelets increased (p<0.0001). Thus, a more rapid reentrant circuit in the superior right atrium drove fibrillatory activity in the remainder of the atrium, as predicted by the "mother wavelet hypothesis." While awaiting in vivo validation, our study indicates that transverse coupling along the crista terminalis may play an important role in the development of atrial fibrillation from atrial flutter. PMID- 11016413 TI - Origins of spiral wave meander and breakup in a two-dimensional cardiac tissue model. AB - We studied the stability of spiral waves in homogeneous two-dimensional cardiac tissue using phase I of the Luo-Rudy ventricular action potential model. By changing the conductance and the relaxation time constants of the ion channels, various spiral wave phenotypes, including stable, quasiperiodically meandering, chaotically meandering, and breakup were observed. Stable and quasiperiodically meandering spiral waves occurred when the slope of action potential duration (APD) restitution was < 1 over all diastolic intervals visited during reentry; chaotic meander and spiral wave breakup occurred when the slope of APD restitution exceeded 1. Curvature of the wave changes both conduction velocity and APD, and their restitution properties, thereby modulating local stability in a spiral wave, resulting in distinct spiral wave phenotypes. In the LRI model, quasiperiodic meander is most sensitive to the Na+ current, whereas chaotic meander and breakup are more dependent on the Ca2+ and K+ currents. PMID- 11016414 TI - Increasing the computational efficiency of a bidomain model of defibrillation using a time-dependent activating function. AB - Realistic simulations of the effects of strong shocks on cardiac muscle require solving the bidomain model, a continuum representation of cardiac tissue by a system of two reaction-diffusion equations. For two- and three-dimensional problems, the computations tend to take a prohibitively long time. This study develops a computationally efficient and accurate approximation of the bidomain model: a "reduced bidomain" model. The approximation is based on the fact that during a strong shock, the extracellular field in the muscle changes only slightly and, therefore, can be approximated by an activating function, following the concept introduced by Rattay (Rattay, F. Analysis of models for external stimulation of axons. IEEE Trans. Biomed. Eng. 33:974-977, 1986). The activating function used here is time-dependent and is computed using an iterative algorithm. The results show that in two spatial dimensions, the "reduced bidomain" model, as implemented in this study, cuts the computational cost by two orders of magnitude while preserving most properties of the "full bidomain" model. It faithfully represents the spatial pattern and the temporal development of the muscle polarization. Consequently, relative errors in the "defibrillation" threshold, the strength of the weakest shock that terminates all electrical activity within 100 ms, are below 10%. PMID- 11016415 TI - Three-dimensional stress and strain in passive rabbit left ventricle: a model study. AB - To determine regional stress and strain distributions in rabbit ventricular myocardium, an anatomically detailed finite element model was used to solve the equations of stress equilibrium during passive filling of the left ventricle. Computations were conducted on a scalable parallel processing computer and performance was found to scale well with the number of processors used, so that stimulations previously requiring approximately 60 min were completed in just over 5 min. Epicardial strains from the model analysis showed good agreement (RMSE = 0.007332) with experimental measurements when material properties were chosen such that cross fiber strain was more heterogeneous than fiber strain, which is also consistent with experimental observations in other species. PMID- 11016416 TI - Generation of an anatomically based three-dimensional model of the conducting airways. AB - An anatomically accurate model of the conducting airways is essential for adequately simulating gas mixing, particle deposition, heat and water transfer, and fluid distribution. We have extended a two-dimensional tree-growing algorithm to three dimensions for generation of a host-shape dependent three-dimensional conducting airway model. Terminal branches in the model are both length limited and volume-supplied limited. A limit is imposed on the maximum possible branch angle between a daughter and parent branch. Comparison of the resulting model with morphometric data shows that the algorithm produces branching and length ratios, path lengths, numbers of branches, and branching angles very close to those from the experimental data. The correlation between statistics from the generated model and those from morphometric studies suggests that the conducting airway structure can be described adequately using a "supply and demand" algorithm. The resulting model is a computational mesh that can be used for simulating transport phenomena. PMID- 11016417 TI - Automatic classification of protein sequences into structure/function groups via parallel cascade identification: a feasibility study. AB - A recent paper introduced the approach of using nonlinear system identification as a means for automatically classifying protein sequences into their structure/function families. The particular technique utilized, known as parallel cascade identification (PCI), could train classifiers on a very limited set of exemplars from the protein families to be distinguished and still achieve impressively good two-way classifications. For the nonlinear system classifiers to have numerical inputs, each amino acid in the protein was mapped into a corresponding hydrophobicity value, and the resulting hydrophobicity profile was used in place of the primary amino acid sequence. While the ensuing classification accuracy was gratifying, the use of (Rose scale) hydrophobicity values had some disadvantages. These included representing multiple amino acids by the same value, weighting some amino acids more heavily than others, and covering a narrow numerical range, resulting in a poor input for system identification. This paper introduces binary and multilevel sequence codes to represent amino acids, for use in protein classification. The new binary and multilevel sequences, which are still able to encode information such as hydrophobicity, polarity, and charge, avoid the above disadvantages and increase classification accuracy. Indeed, over a much larger test set than in the original study, parallel cascade models using numerical profiles constructed with the new codes achieved slightly higher two-way classification rates than did hidden Markov models (HMMs) using the primary amino acid sequences, and combining PCI and HMM approaches increased accuracy. PMID- 11016418 TI - Bayesian identification of a population compartmental model of C-peptide kinetics. AB - When models are used to measure or predict physiological variables and parameters in a given individual, the experiments needed are often complex and costly. A valuable solution for improving their cost effectiveness is represented by population models. A widely used population model in insulin secretion studies is the one proposed by Van Cauter et al. (Diabetes 41:368-377, 1992), which determines the parameters of the two compartment model of C-peptide kinetics in a given individual from the knowledge of his/her age, sex, body surface area, and health condition (i.e., normal, obese, diabetic). This population model was identified from the data of a large training set (more than 200 subjects) via a deterministic approach. This approach, while sound in terms of providing a point estimate of C-peptide kinetic parameters in a given individual, does not provide a measure of their precision. In this paper, by employing the same training set of Van Cauter et al., we show that the identification of the population model into a Bayesian framework (by using Markov chain Monte Carlo) allows, at the individual level, the estimation of point values of the C-peptide kinetic parameters together with their precision. A successful application of the methodology is illustrated in the estimation of C-peptide kinetic parameters of seven subjects (not belonging to the training set used for the identification of the population model) for which reference values were available thanks to an independent identification experiment. PMID- 11016419 TI - Shear stress differentially regulates PGHS-1 and PGHS-2 protein levels in human endothelial cells. AB - The secretion of prostacyclin (PGI2) by endothelial cells is regulated by shear stress. Prostaglandin H synthase (PGHS) is considered to be a key limiting enzyme in the synthesis of PGI2 from arachidonic acid. Endothelial cells were cultured in the presence of 4, 15, or 25 dyn/cm2 shear stress using a parallel plate flow chamber to assess the effect of shear stress on both PGHS isoforms, PGHS-1 and PGHS-2. In cells exposed to 4, 15, or 25 dyn/cm2 shear stress PGHS-1 and PGHS-2 protein levels initially decreased. The decrease was followed by a sustained increase for PGHS-1 but only a transient increase for PGHS-2. The duration of the PGHS-2 increase depended on the magnitude of the shear stress. The effect of altering shear stress levels on PGHS protein levels in cells preconditioned to either 4, 15, or 25 dyn/cm2 shear stress for 48 h was also studied. Changing shear stress levels effected PGHS-2 but not PGHS-1. Increases in shear stress levels from 4 to 15 or 25 dyn/cm2 caused a decrease in PGHS-2. In contrast, decreases in shear stress levels from 15 or 25 to 4 dyn/cm2 caused PGHS-2 to increase. There was a continual decrease in PGHS-2 when the shear stress was changed from 15 to 25 or 25 to 15 dyn/cm2. In summary, the regulation of PGHS-2 by shear stress is dependent upon the magnitude of the shear stress, whereas the regulation of PGHS-1 protein levels seems to be independent of the shear stress magnitude. The regulation of PGHS-1 and PGHS-2 protein levels by shear stress indicates that these proteins play an important role in the maintenance of cardiovascular homeostasis as regulators of PGI2 production. PMID- 11016420 TI - Preface and overview: genetics of SLE; a sine qua non for identification. AB - Clinical manifestations of systemic lupus erythematosus (SLE) are extremely diverse and variable, mainly because SLE is a multi-factorial disease. Variable combinations of contributing genes at multiple loci in individual patients probably result in diverse disease phenotypes. Genes that predispose to SLE are undoubtedly related to key events in pathogenesis, and may involve a variety of genes in immune system. These genes are currently unidentified, mostly because of the complexity of multi-factorial inheritance. Recently, the application of the polymerase chain reaction and the availability of maps of microsatellites have facilitated a genome-wide scan to define the number and locations of genes for complex traits. However, extensive genetic heterogeneities and polymorphisms and complex modes of inheritance of disease phenotypes have delayed completion of a genome-wide analysis of susceptibility loci for human SLE. Since many SLE susceptibility genes show low penetrance, several hundred affected sibpairs are assumed to be necessary to show linkages for many of the contributing loci. In this respect, studies of polymorphisms and functions of candidate genes suggested based on studies on murine models may contribute to studies on SLE patients and their relatives. Major genetic loci have been mostly identified in SLE-prone mouse strains. Nevertheless, identity of natures, functions and roles in the pathogenesis of SLE remains undetermined. Considering the possibility of clustering of susceptibility loci in a particular chromosomal interval, the final goal for identification of susceptibility genes will largely depend on the generation of mutant SLE-prone mice with homologous recombination of the potential target gene. Extensive reviews collected here are expected to form the basis for identification of target genes and for clarification of the genetic mechanisms underlying SLE. PMID- 11016421 TI - Genetic studies of human lupus in families. AB - Systemic lupus erythematosus (SLE) is a complex multigenic disease in which the contributing genetic systems are being rapidly identified. Most of the currently recognized genes have been discovered from case-control association studies, but, increasingly, family linkage studies are being employed to confirm previous genetic associations, to examine their relative contributions, and to identify new susceptibility loci. Most of the loci identified thus far appear to contribute only modest effects on susceptibility overall but rather influence more strongly disease expression and/or severity. MHC class II alleles, for example, seem to show only weak linkage to SLE itself but instead mediate specific T cell driven pathogenic autoantibodies which produce many of the clinical disease features, similar to their effects in many other autoimmune diseases. On the other hand, complete and partial hereditary deficiencies of early complement components are more lupus-specific. Homozygous complement deficiencies, while powerful risk factors, are rare causes of lupus and heterozygous deficiencies exert only modest effects on susceptibility. Other genes, such as low-binding IgG Fc receptor alleles (FcgammaIIa and FcgammaIIIa), appear to promote nephritis by modifying the efficiency of immune complex clearance. A variety of cytokine genes appear also to promote severity, including those for TNFalpha, IL-10, IL1 receptor antagonist, and perhaps others (IL-6, IL 4 and TNFalpha receptor). Family studies and recent genome-wide scans in lupus and other autoimmune diseases support the likelihood that some susceptibility loci, as yet unidentified, predispose to several or many autoimmune diseases. Only thorough the identification and elucidation of function of these many genes is the pathogenic picture of lupus likely to be complete. PMID- 11016422 TI - Lupus susceptibility genes on human chromosome 1. AB - During the past five years, there has been an intense interest in studying candidate susceptibility genes for systemic lupus erythematosus (SLE). Many such studies have been focused on candidates located on chromosome 1, demonstrating association of certain genetic variants with SLE. Some of the tested candidate genes were chosen because they encode molecules with relevant immunological functions that may play a role in the pathogenesis of SLE. More recently, the identification of genomic segments linked to SLE has suggested novel positional candidate genes. Thus far, there is considerable evidence supporting that multiple genes on this chromosome contribute to the development and expression of SLE. This review highlights the genetic loci located on chromosome 1 that have recently been associated with SLE. These include loci encoding the tumor necrosis factor receptor 2 (TNFR2), complement component C1q, Fcgamma receptors, T cell receptor zeta chain, interleukin-10 (IL-10), poly (ADP-ribose) polymerase (PARP), and HRES-1. PMID- 11016423 TI - Practical approaches to determining disease-susceptible loci in multigenic autoimmune models. AB - Linkage analysis using polymorphic DNA markers has paved the way toward the identification of genes responsible for rare recessive traits and for the susceptibility to certain tumors in humans. However, genetic susceptibility to common diseases, including systemic autoimmune diseases, is difficult to determine, hence has remained a challenging problem in the field of molecular genetics. Elucidation of multiple quantitative trait loci that predispose individuals to multi-phenotypic systemic autoimmune disease requires formidable research efforts, and there is a growing consensus that mouse models are required. This review provides a guide to methods that can be used in linkage studies of autoimmune mice. Mouse studies in relation to recent advances in bio informatics are also discussed. PMID- 11016424 TI - Genetics of systemic autoimmunity in mouse models of lupus. AB - Systemic lupus erythematosus (SLE) is inherited as a complex polygenic trait, involving genetic, environmental and stochastic factors. Although definition of these etiologic processes has been elusive, solid progress has been made toward elucidating the genetic basis for susceptibility. Herein, we summarize our genome wide mapping effort that has defined loci for component phenotypes for lupus prone NZB, NZW, MRL-Fas(lpr) and BXSB strains. With this framework in place, identification of the specific genetic alterations and mechanisms is now proceeding through the generation of interval congenic lines, precise mapping and screening of candidate genes. In addition to this approach, transgenic and gene knockout studies have begun to identify genes that can induce or modify autoimmunity in nonautoimmune and lupus-prone background mice, including studies by us and others on Th1 and Th2 cytokine genes in lupus. It is apparent that a diversity of genes and mechanisms can independently or in combination promote systemic autoimmunity in mice. This complexity, which is also observed in human lupus, emphasizes the importance of using experimental and less complex mouse models to define these processes, a tactic that has already yielded new insights. With current technologies and the anticipated definition of mammalian genomes, identification of genes predisposing to lupus and elucidation of processes critical for disease pathogenesis appear within grasp. PMID- 11016425 TI - Genetic aspects of inherent B-cell abnormalities associated with SLE and B-cell malignancy: lessons from New Zealand mouse models. AB - Genes that predispose to SLE are closely related to key events in pathogenesis of this disease. As much of the pathology can be attributed to high affinity autoantibodies and/or their immune complexes, some of the genes may exert effects in the process of emergence, escape from tolerance mechanisms, activation, clonal expansion, differentiation, class switching and affinity maturation of self reactive B cells. A number of growth and differentiation factors and signaling molecules, including positive and negative regulators, are involved in this process. Genetic variations associated with functional deficits in some of such molecules can be involved in the susceptibility for SLE. As is the case with SLE, hereditary factors play significant roles in the pathogenesis of B cell chronic lymphocytic leukemia (B-CLL). Patients with B-CLL or their family members frequently have immunological abnormalities, including those associated with SLE. It is suggested that certain genetically determined regulatory abnormalities of B cells may be a crossroad between B-CLL and SLE. A thorough understanding of the genetic pathways in B cell abnormalities leading to either SLE or B-CLL is expected to shed light on their association. New Zealand mouse strains are pertinent laboratory models for these studies. Chromosomal locations of several major genetic loci for abnormal proliferation, differentiation and maturation of B cells and relevant candidate genes, located in close proximity to these intervals and potentially related to the SLE pathogenesis, have been identified in these mice. Further studies make for a wider knowledge and understanding of the pathogenesis of SLE and related B-cell malignancy. PMID- 11016426 TI - Lessons from the NZM2410 model and related strains. AB - SLE susceptibility requires the interplay of an unknown number of genes and equally unidentified triggering events. The past few years have seen significant advances in our understanding of SLE susceptibility through the genetic analysis of murine models. The NZM2410 strain, which is derived from the NZB/WF1 model has played a significant role in these advances. The main advantages presented by this strain over other models are the genetic homozygozity at all loci and an highly penetrant early onset lupus nephritis in both males and females, indicating that the strongest BWF1 susceptibility loci were retained in NZM2410. After identification of NZM2410 susceptibility loci via linkage analyses, congenic strains have been derived in order to convert a polygenic system into a series of monogenic traits. These congenic strains have been analyzed in an integrated process which has provided simultaneously 1) novel functional characterization of the Sle susceptibility loci, 2) high resolution genetic maps that will lead to the identification of the corresponding susceptibility genes by either candidate locus or positional cloning, and 3) insights into the mechanisms by which these loci interact to produce systemic autoimmunity with fatal end organ damage. PMID- 11016427 TI - Lessons from BXSB and related mouse models. AB - The BXSB murine strain spontaneously develops an autoimmune syndrome with features of systemic lupus erythematosus (SLE) that affects males much earlier than females, due to the presence of an as yet unidentified mutant gene located on its Y chromosome, designated Yaa (Y-linked autoimmune acceleration). The Yaa gene by itself is unable to induce significant autoimmune responses in mice without an apparent SLE background, while it can induce and accelerate the development of an SLE in combination with autosomal susceptibility alleles present in lupus-prone mice. Although the genes encoded within or closely linked to the MHC locus play an important role in the development or protection of SLE, the MHC effect can be completely masked by the presence of the Yaa gene in mice highly predisposed to SLE. The role of the Yaa gene for the acceleration of SLE is apparently two-fold; it enhances overall autoimmune responses against autoantigens to which mice respond relatively weakly, and promotes Th 1 responses against autoantigens to which mice respond relatively well, leading to the production of more pathogenic autoantibodies, i.e., FcgammaR-fixing IgG2a and cryoglobulin IgG3 autoantibodies. Yaa+ - Yaa- double bone marrow chimera experiments revealed that the Yaa defect is expressed in B cells, but not in T cells, and that T cells from non-autoimmune mice are capable of providing help for autoimmune responses by collaborating Yaa+ B cells. We speculate that the Yaa defect may decrease the threshold for antigen receptor-dependent stimulation, leading to the triggering and excessive stimulation of autoreactive T and B cells. PMID- 11016428 TI - Genetic basis of the complex pathological manifestations of collagen disease: lessons from MRL/lpr and related mouse models. AB - The pathological findings in collagen disease including systemic lupus erythematosus show complex lesions such as glomerulonephritis, systemic vasculitis, polyarthritis, sialoadenitis, etc. Moreover, some cases of collagen disease are categorized into overlapping syndromes. It is still controversial whether such diversity and similarity of pathological manifestations among the collagen disease depends on ambiguity in diagnosis or is an intrinsic quality of the collagen diseases themselves. In this paper, we reviewed this subject focusing on a series of our genetic studies of murine models of collagen disease, MRL strains of mice with a deficit in Fas-mediated apoptosis, which spontaneously develop glomerulonephritis, systemic vasculitis, polyarthritis and sialoadenitis. We observed that each lesion was controlled by a different set of genes and they appeared to act in an additive manner on the development of each lesion. We conclude that various disease categories in collagen disease will be a result of the combination of polygenes. PMID- 11016429 TI - Environmental information systems for the control of arthropod vectors of disease. AB - Over the last decade, remote sensing technologies and geographical information systems have moved from the research arena into the hands of vector control specialists. This review explains remote sensing approaches and spatial information technologies used for investigations of arthropod pests and vectors of diseases affecting humans and livestock. Relevant applications are summarized with examples of studies on African horse sickness vector Culicoides midges (Diptera: Ceratopogonidae), malaria vector Anopheles and arbovirus vector culicine mosquitoes (Diptera: Culicidae), leishmaniasis vector Phlebotomus sandflies (Diptera: Psychodidae), trypanosomiasis vector tsetse (Diptera: Glossinidae), loaiasis vector Chrysops (Diptera: Tabanidae), Lyme disease vector Ixodes and other ticks (Acari: Ixodidae). Methods and their uses are tabulated and discussed with recommendations for efficiency, caution and progress in this burgeoning field. PMID- 11016430 TI - African horse sickness epidemiology: vector competence of south african Culicoides species for virus serotypes 3, 5 and 8. AB - The oral susceptibilities of 17 Culicoides species to infection with African horse sickness virus (AHSV) serotypes 3, 5 and 8 were determined by feeding field collected midges on AHSV infected horse blood. The mean titres of virus in the bloodmeals for the three serotypes of AHSV were between 5.7 and 6.5 log10 TCID50/ml. Virus was detected, after 10 days incubation at 23.5 degrees C, in the Culicoides imicola Kieffer (Diptera: Ceratopogonidae) that had fed on blood containing AHSV 5 (8.5%) and 8 (26.8%), and in the Culicoides bolitinos Meiswinkel that had fed on AHSV 3 (3.8%), 5 (20.6%) and 8 (1.7%). Although 44.4% of the C. imicola were shown to have ingested AHSV 3 immediately after feeding, no virus was detected in 96 C. imicola after incubation. The relatively high titres of virus recorded in individual midges of both species after 10 days incubation suggested a fully disseminated infection. Previously, C. imicola was considered to be the only field vector of AHSV in Africa. Identifying C. bolitinos as a potential vector for AHSV is an important finding, which if proven will have a significant impact on our understanding of the epidemiology of AHS. No AHSVs could be detected in the other 15 species of Culicoides assayed, which suggests that some of the southern African Culicoides species are refractory to AHSV infection. However, further work with larger numbers of each species will be necessary to confirm this observation. PMID- 11016431 TI - Salivary proteins and glycoproteins in phlebotomine sandflies of various species, sex and age. AB - Salivary gland proteins were studied in sandflies (Diptera: Psychodidae: Phlebotominae) by electrophoretic techniques. In Phlebotomus duboscqi Neveu Lemaire the protein concentration was about 30 times higher in females than in males. SDS PAGE revealed eight major bands of 29-62 kDa in salivary gland extracts (SGE) from females, whereas only one band of 57kDa was detected in males. The number of protein components in SGE gradually increased with the age of females. In P. papatasi (Scopoli) the typical electrophoretic pattern was reached in 3-5 days after imago emergence, depending on the temperature at which females were maintained. All major protein components of the female SGE were present in the content of glands. Female SGE were compared in seven colonies of five Phlebotomus and Lutzomyia species; electrophoretic profiles distinguished between species and even between colonies of different geographical origin. In general, the highest variability of major protein components was observed in the 38-48kDa region. Four colonies of the subgenus Phlebotomus (P. duboscqi and P. papatasi) possessed common mobility polypeptides, the highest similarity was found between two colonies of P. papatasi. Other species tested significantly differed, specific prominent bands of 33, 35 and 38kDa were found in P. halepensis Theodor, P. perniciosus Newstead and Lutzomyia longipalpis (Lutz & Neiva), respectively. Glycoproteins in SGE of Lu. longipalpis and P. duboscqi females were identified and analysed using blotting with five lectin conjugates. Specific reaction of lectins ConA and WGA revealed the complex type of N-glycans in the 48 and 53-54kDa glycoproteins present in both species. Similar glycosylation was detected in species-specific bands of the 57-60 and 65-67 kDa in P. duboscqi and Lu. longipalpis, respectively. The high mannose type of glycosylation was found in the 20 and 39 kDa polypeptides of Lu. longipalpis and the 40-42 kDa polypeptides of P. duboscqi. Innate lectin activity specific for aminosugars was detected in SGE of P. duboscqi females using haemagglutination tests with rabbit erythrocytes. PMID- 11016432 TI - Comparison of carbon dioxide-baited trapping systems for sampling outdoor mosquito populations in Tanzania. AB - For collecting mosquitoes (Diptera: Culicidae) the outdoor catching efficiency of four types of trapping devices baited with carbon dioxide (CO2, 300 ml/ min) was evaluated and compared in two areas of Tanzania. The types of traps employed were: the CDC miniature trap with the incandescent light bulb switched on or off; electric nets (ENT) and a Counterflow Geometry (CFG) trap. In Njage, southeast Tanzania, Anopheles gambiae Giles sensu stricto was the most abundant of the seven mosquito species obtained, comprising of 74.3% of the total number caught (n=2,171). In Muheza, north-east Tanzania, Culex quinquefasciatus Say was the predominant species (90.9%) among 1,080 caught. At both localities the CFG trap was superior to the CDC trap with light-on or light-off for sampling both An. gambiae and Cx. quinquefasciatus. Efficiency of the CFG trap and ENT were similar for sampling these species of mosquitoes (P > 0.05). However, ENT was superior to the CDC trap with light-off for collecting both species. Significantly more (P < 0.05) Cx. quinquefasciatus were obtained by the CDC trap with light-off than with light-on, especially outdoors. It is concluded that both ENT and the CFG are effective tools for sampling populations of An. gambiae and Cx. quinquefasciatus outdoors. PMID- 11016433 TI - Genetic differentiation of Anopheles claviger s.s. in France and neighbouring countries. AB - An investigation of polymorphism of 11 autosomal and one sex-linked allozyme loci was made on 18 samples of Anopheles claviger Meigen (Diptera: Culicidae) from localities across France and neighbouring sites in Germany and Switzerland, plus one sample of Anopheles petragnani Del Vecchio from the French Pyrenees. Genetic differentiation between these two sibling species was confirmed (Nei genetic distance 0.33-0.44) and two genetically distinct groups of populations were identified within An. claviger. These two forms of An. claviger showed contiguous geographical distributions, Group I found across western and Central France, Group II in eastern France and nearby parts of Germany and Switzerland. The two groups were in contact in a region near the Rhone Valley where two intermediate samples were found. The taxonomic significance of this finding is discussed in the context of the recent climatic history of Europe and in relation to the vector potential of each member of the An. claviger complex. PMID- 11016434 TI - Cattle breed-variation in infestation by the horn fly Haematobia irritans. AB - A study was carried out to assess the resistance of pure and cross-bred groups of cattle to the horn fly Haematobia irritans (Linnaeus) (Diptera: Muscidae) in northern Argentina. Pure-bred cattle were Criolla, Iberian Bos taurus Linnaeus (Artiodactyla: Bovidae) and Nellore, Bos indicus Linnaeus (Artiodactyla: Bovidae). Cross-bred cattle were Hereford, British B. taurus (34%) X Nellore (66%) and Hereford (66%) X Nellore (34%). All were heifers and animals were maintained in two groups, each containing a mixture of pure and cross-breeds. The lowest fly numbers were found on Criolla heifers and the highest on Hereford X Nellore cross-breeds. However, it could not be determined from this study whether this was a consequence of breed and/or size, as Criolla heifers were lighter than the corresponding Hereford X Nellore heifers. Fly numbers on the heifers followed an approximately negative binomial distribution. However, the ranking of individual animals in their level of infestation within subgroups was not consistent. Hence, culling the most infested heifers on any given date would at best give only a small improvement in H. irritans control. PMID- 11016435 TI - Role of small mammals in the persistence of Louping-ill virus: field survey and tick co-feeding studies. AB - Louping-ill (LI) is a tick-borne viral disease of red grouse, Lagopus lagopus scoticus Lath. (Tetraonidae: Galliformes), and sheep, Ovis aries L. (Bovidae: Artiodactyla), that causes economic loss to upland farms and sporting estates. Unvaccinated sheep, grouse and mountain hares, Lepus timidus L. (Leporidae: Lagomorpha), are known to transmit LI virus, whereas red deer, Cenrus elaphus L. (Cervidae: Artiodactyla), and rabbits, Oryctolagus cuniculus L. (Leporidae: Lagomorpha), do not. However, the role of small mammals is unknown. Here, we determine the role of small mammals, in particular field voles, Microtus agrestis L. (Muridae: Rodentia), in the persistence of LI virus on upland farms and sporting estates in Scotland, using field sampling and non-viraemic transmission trials. Small mammals were not abundant on the upland sites studied, few ticks were found per animal and none of the caught animals tested seropositive to LI virus. Laboratory trials provided no evidence that small mammals (field voles, bank voles, Clethrionomys glareolus L. (Muridae: Rodentia), and wood mice, Apodemus sylvaticus L. (Muridae: Rodentia), can transmit LI virus between cofeeding ticks and, in the field, LI virus was prevalent only in areas with known LI virus competent hosts (grouse, mountain hares or unvaccinated sheep) and absent elsewhere. In contrast to the case of tick-borne encephalitis (TBE) virus in Europe, it is concluded that small mammals seem to be relatively unimportant in LI virus persistence. PMID- 11016436 TI - Responses of Glossina austeni to sticky panels and odours. AB - The responses of male tsetse Glossina austeni Newstead (Diptera: Glossinidae) towards blue and white sticky legged panels, baited with odour attractants, and towards modified panels were studied in the Jozani forest of Unguja Island, Zanzibar. Increasing the height of the body of a standard panel from 30 to 60 or 90 cm, increased the catch two-fold. Increasing the height of the legs (from 15 to 60 or 120 cm) or raising the device more than 5 cm above the ground reduced the catch significantly. The legs of the panels were the preferred landing sites of the flies, irrespective of the height of the body of the panel. Acetone (300 mg/h) combined with cow urine (60-130 mg/h) significantly increased the catches two- to threefold during the rainy season, but not during the dry season. Acetone had no effect during the dry season and its effect during the rainy season was less consistent. There was no effect of octenol (2.5-12.5 mg/h), used alone or in combination with acetone. Likewise, the catch did not increase through the addition of cow sebum, pig urine (60-860 mg/h), pig urine combined with acetone and octenol. The observed seasonal differences in the response of G. austeni towards odours are discussed in relation to host location strategies. PMID- 11016437 TI - Feeding behaviour of Glossina pallidipes and g. morsitans centralis on Boran cattle infected with trypanosoma congolense or T. vivax under laboratory conditions. AB - In field studies, tsetse flies (Diptera: Glossinidae) feed more successfully on cattle infected with Trypanosoma congolense Broden (Kinetoplastida: Trypanosomatidae) than on cattle infected with T. vivax Ziemann or uninfected cattle. Here we describe the first laboratory investigation of this phenomenon. In the first experiment, caged Glossina pallidipes Austen were fed for 1 and 5 min on a Boran steer infected with T. congolense clone IL 1180 and on an uninfected steer. Feeding success was recorded in this way five times over several weeks. The same protocol was subsequently used in three additional experiments with the following combinations: G. pallidipes and a steer infected with T. vivax stock IL 3913, G. morsitans centralis Machado and a steer infected with T. congolense, and G. morsitans centralis and a steer infected with T. vivax. The four experiments were replicated once, making eight experiments in total. In three experiments there was increased tsetse feeding success, measured at 1 min, after a steer became infected (T. congolense, two experiments and T. vivax, one experiment). Analysis of all data combined found no significant differences in tsetse feeding success on the different groups of cattle prior to infection, but after infection tsetse feeding success was significantly greater on the infected cattle (P< 0.001). Trypanosoma congolense infection led to a greater increase in tsetse feeding success than T. vivax infection. The increase in feeding success was not related to changes in the level of anaemia, skin surface temperature or parasitaemia. A possible explanation is the effects of trypanosome infection on cutaneous vasodilation and/or blood clotting in infected cattle. When allowed to feed for 5 min, nearly all tsetse engorged successfully and effects of cattle infection on feeding success were not found. PMID- 11016438 TI - Responses of the biting midge Culicoides impunctatus to acetone, CO2 and 1-octen 3-ol in a wind tunnel. AB - Responses of host-seeking female Culicoides impunctatus (Goetghebuer) (Diptera: Ceratopogonidae) to acetone, carbon dioxide and 1-octen-3-ol were measured in a wind tunnel. Carbon dioxide, presented as a filamentous plume, increased upwind flight in a dose-dependent manner, up to 0.09% concentration. A homogenous CO2 plume elicited similar upwind responses at concentrations up to 0.09%, whereas higher plume concentrations (> 0.1%) induced erratic responses with a suppression of upwind flight. Bovine equivalent concentrations of acetone (1.5 x 10(-6)g/l) and 1-octen-3-ol (1.3 x 10(-8)g/) failed to induce any significant upwind response when tested alone. In the presence of CO2, however, 1-octen-3-ol showed highly significant increases in upwind responses at concentrations of 1.3 x 10( 1) - 10(-8)g/l. Mixtures of CO2+ acetone also enhanced upwind flight at 1.5 x 10( 9)g/l. High tunnel concentrations of both 1-octen-3-ol and acetone inhibited upwind responses. These findings are discussed in relation to host finding by C. impunctatus and known mechanisms by which upwind flight is initiated and arrested at high odour concentrations. PMID- 11016439 TI - Differential ecology of Anopheles punctulatus and three members of the Anopheles farauti complex of mosquitoes on Guadalcanal, Solomon Islands, identified by PCR RFLP analysis. AB - From a series of larval collections made across northern Guadalcanal during the dry season, October-November 1997, four members of the Anopheles punctulatus group of mosquitoes (Diptera: Culicidae) were identified using PCR-RFLP analysis. Anopheline larvae were found in 54/57 (95%) of the sites sampled, comprising An. farauti Laveran sensu stricto (32 sites), An. farauti species no. 2 (39 sites), An. farauti no. 7 (36 sites) and An. punctulatus Donitz (10 sites). Anopheles punctulatus occurred only on the coastal plain, where it was associated with the more transient sites. Anopheles farauti sensu lato was more widespread throughout the survey region, with similar proportions of all three sibling species in both transient and permanent sites. Two members of the An. farauti complex, An. farauti s.s. and species no. 2, were found in brackish water. All breeding sites of An. punctulatus were cohabited by An. farauti s.l., sometimes by all three sibling species. Anopheles farauti s.s. was the only species collected on human bait, with a much higher biting rate early in the evening (57 bites/human/hour at 18.30-20.00 hours) than later (0.8 bites/human/hour at 21.00-24.00 hours). PMID- 11016441 TI - Phosphoglucomutase and trehalase isoenzymes of Venezuelan simulium vectors of Onchocerca volvulus. AB - Phosphoglucomutase (PGM) and trehalase (Tre) isoenzymes of five species of Simulium blackflies (Diptera: Simuliidae), vectors of onchocerciasis in Venezuela, were investigated by means of a portable electrophoresis field kit. Tre differed between S. incrustatum and S. oyapockense s.l. Electrophoretic variation of Tre in other members of the S. amazonicum and S. incrustatum groups merit further investigation. PGM appears to be more useful for separating populations within species complexes. Multiple populations and/or seasonal changes in population structure of S. guianense s.l., S. exiguum s.l. and S. metallicum s.l. were inferred from elecrophoretic variation of PGM. PMID- 11016440 TI - Feeding and survival of Culicoides sonorensis on cattle treated with permethrin or pirimiphos-methyl. AB - The persistence of permethrin (5% a.i.) and pirimiphos-methyl (27% a.i.), applied to the dorsum of calves in the field against Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae), was estimated using a hair-blood-feeding bioassay in the laboratory. Hair clippings were taken before treatment and 3, 7, 14, 21, 28, 42 and 56 days after treatment from the dorsum, side and belly of treated and control calves. Laboratory-reared insects were allowed to feed through thin hair layers and a parafilm membrane on sheep blood warmed using a water-jacketed feeder. Some intoxication after exposure to hair was noted up to 28 days after treatment with permethrin and up to 14 days after treatment with pirimiphos-methyl. Hair from the dorsum caused more intoxication for a longer period than hair from other body regions. Permethrin and pirimiphos-methyl applied to the back did not significantly reduce overall engorgement (body regions pooled) after treatment. Permethrin residues on hair remained far higher on the back than other body regions and were related to insect intoxication and reduction in engorgement in the laboratory. Residues on belly hair never exceeded 12p.p.m. and did not result in significantly reduced feeding at any time. Engorged insects that exhibited sublethal intoxication from feeding through permethrin-treated hair did recover and matured numbers of eggs comparable to controls. Field trials using treated and control calves and enclosure nets showed that dorsal applications of 5% permethrin were not effective in reducing engorgement, despite some intoxication. Vacuum samples from a calf showed that C. sonorensis fed primarily on the belly. A 0.2% permethrin application on the belly (250 ml) did result in > 80% reduction of C. sonorensis in the enclosure nets at 3 and 7 days after treatment, but activity had subsided by 10 days after treatment. The utility of insecticidal treatments for suppression of this vector is discussed. PMID- 11016442 TI - The ixodid tick species attaching to domestic dogs and cats in Great Britain and Ireland. AB - The species of ixodid ticks, attached to dogs and cats presented to veterinary practices in Great Britain and Ireland were identified. Most host animals carried only one tick species with Ixodes ricinus Linne (Acari: Ixodidae) being the most common, identified on 52% of animals, Ixodes hexagonus Leach (Acari: Ixodidae) the second most common (on 39%) and Ixodes canisuga Johnston (Acari: Ixodidae) the third most common (on 11%). A significantly higher proportion of dogs than cats carried I. ricinus, while I. hexagonus was more frequently carried by cats. One animal carried a single specimen of Haemaphysalis punctata Canestrini & Fanzago (Acari: Ixodidae), one carried a Dermacentor reticulatus Fabricius (Acari: Ixodidae) but none carried Rhipicephalus sanguineus Latreille (Acari: Ixodidae). This indicates that the latter two species, vectors of 'exotic' tick borne diseases, remain at low densities in Great Britain and Ireland. Retrospective information on exposure of the animals to different habitats and geographic regions was collected by questionnaire and subject to contingency table and logistic regression analysis. Woodlands and moorlands were habitats significantly associated with I. ricinus attachment. Exposure to urban parks was significantly associated with I. hexagonus attachment and exposure to boarding kennels and catteries was significantly associated with I. canisuga attachment. Ixodes hexagonus, rather than I. ricinus, was the ixodid tick species most likely to be encountered by urban populations of dogs and cats and, by inference, possibly also humans. The implications of these findings, for the transmission of tick-borne pathogens to dogs, cats and humans are discussed. PMID- 11016443 TI - Density and dispersal of the loaiasis vector Chrysops dimidiata in southern Cameroon. AB - By mark-release-recapture experiments, we assessed the density of loaisis vectors, Chrysops dimidiata Wulp plus some Chrysops silacea Austen (Diptera: Tabanidae) and estimated their range of flight in the secondary forest of southern Cameroon. In 1993, the release point was at the centre of the study area and recapture points were at 1,100 m radius. In 1994, releases were on the periphery of the study area and recapture sites were 400-8,000 m from the release points. Results were concordant and showed Chrysops female densities of 785-3,682 flies/ km2. The theoretical flight range was < 6,000 m, with a maximum distance of 4,500 m observed. These results are considered promising for the use of vector control methods against loaiasis. PMID- 11016444 TI - GAD65-specific autoantibodies enhance the presentation of an immunodominant T cell epitope from GAD65. AB - GAD65 autoantibodies (GAD65Ab) are highly prevalent in type 1 diabetes, but their functional role in the pathogenesis of the disease and their relationship to T cell reactivity to GAD65 is still unclear. We tested the hypothesis that GAD65Ab modulate presentation of GAD65 to T-cells. T-cell hybridoma T33.1, which recognizes the GAD65 274-286 epitope in the context of HLA-DRB 1*0401, was incubated with antigen-presenting cells exposed to recombinant human GAD65 alone or complexed with GAD65Ab' or GAD65Ab- sera. Stimulation of the T33.1 hybridoma was greatly enhanced by multiple GAD65Ab+ sera. The enhancement effect was most prominent with sera from patients with high GAD65 autoantibody levels. Sera from GAD65Ab- subjects had no effect. The correlation between T-cell stimulation and GAD65Ab levels was not absolute, suggesting that other variables such as autoantibody recognition of different regions of GAD65 and variable effects on processing of the 274-286 epitope may contribute. Uptake of antibody-complexed GAD65 was Fc receptor (FcR)-mediated because the enhancement of presentation was inhibited by monoclonal antibodies against FcR. Our results support the hypothesis that GAD65Ab modulate presentation of GAD65 to T-cells. Increased antigen uptake and heterogeneity in the autoantibody specificity may provide a mechanism for antibody-facilitated T-cell response influencing the progression of type 1 diabetes. PMID- 11016445 TI - Identification, cloning, and heterologous expression of a mammalian fructosamine 3-kinase. AB - Fructosamines are thought to play an important role in the development of diabetic complications. Little is known about reactions that could metabolize these compounds in mammalian tissues, except for recent indications that they can be converted to fructosamine 3-phosphates. The purpose of the present work was to identify and characterize the enzyme responsible for this conversion. Erythrocyte extracts were found to catalyze the ATP-dependent phosphorylation of 1-deoxy-1 morpholinofructose (DMF), a synthetic fructosamine. The enzyme responsible for this conversion was purified approximately 2,500-fold by chromatography on Blue Sepharose, Q Sepharose, and Sephacryl S-200 and shown to copurify with a 35,000 M(r) protein. Partial sequences of tryptic peptides were derived from the protein by nanoelectrospray-ionization mass spectrometry, which allowed for the identification of the corresponding human and mouse cDNAs. Both cDNAs encode proteins of 309 amino acids, showing 89% identity with each other and homologous to proteins of unknown function predicted from the sequences of several bacterial genomes. Both proteins were expressed in Escherichia coli and purified. They were shown to catalyze the phosphorylation of DMF, fructoselysine, fructoseglycine, and fructose in order of decreasing affinity. They also phosphorylated glycated lysozyme, though not unmodified lysozyme. Nuclear magnetic resonance analysis of phosphorylated DMF and phosphorylated fructoseglycine showed that the phosphate was bound to the third carbon of the 1-deoxyfructose moiety. The physiological function of fructosamine-3-kinase may be to initiate a process leading to the deglycation of fructoselysine and of glycated proteins. PMID- 11016446 TI - Portal glucose infusion in the mouse induces hypoglycemia: evidence that the hepatoportal glucose sensor stimulates glucose utilization. AB - To analyze the role of the murine hepatoportal glucose sensor in the control of whole-body glucose metabolism, we infused glucose at a rate corresponding to the endogenous glucose production rate through the portal vein of conscious mice (Po mice) that were fasted for 6 h. Mice infused with glucose at the same rate through the femoral vein (Fe-mice) and mice infused with a saline solution (Sal mice) were used as controls. In Po-mice, hypoglycemia progressively developed until glucose levels dropped to a nadir of 2.3 +/- 0.1 mmol/l, whereas in Fe mice, glycemia rapidly and transiently developed, and glucose levels increased to 7.7 +/- 0.6 mmol/l before progressively returning to fasting glycemic levels. Plasma insulin levels were similar in both Po- and Fe-mice during and at the end of the infusion periods (21.2 +/- 2.2 vs. 25.7 +/- 0.9 microU/ml, respectively, at 180 min of infusion). The whole-body glucose turnover rate was significantly higher in Po-mice than in Fe-mice (45.9 +/- 3.8 vs. 37.7 +/- 2.0 mg x kg(-1) x min)-1), respectively) and in Sal-mice (24.4 +/- 1.8 mg x kg(-1) x min(-1)). Somatostatin co-infusion with glucose in Po-mice prevented hypoglycemia without modifying the plasma insulin profile. Finally, tissue glucose clearance, which was determined after injecting 14C-2-deoxyglucose, increased to a higher level in Po-mice versus Fe-mice in the heart, brown adipose tissue, and the soleus muscle. Our data show that stimulation of the hepatoportal glucose sensor induced hypoglycemia and increased glucose utilization by a combination of insulin dependent and insulin-independent or -sensitizing mechanisms. Furthermore, activation of the glucose sensor and/or transmission of its signal to target tissues can be blocked by somatostatin. PMID- 11016447 TI - Glucose sensing by the hepatoportal sensor is GLUT2-dependent: in vivo analysis in GLUT2-null mice. AB - In the preceding article, we demonstrated that activation of the hepatoportal glucose sensor led to a paradoxical development of hypoglycemia that was associated with increased glucose utilization by a subset of tissues. In this study, we tested whether GLUT2 plays a role in the portal glucose-sensing system that is similar to its involvement in pancreatic beta-cells. Awake RIPGLUT1 x GLUT2-/- and control mice were infused with glucose through the portal (Po-) or the femoral (Fe-) vein for 3 h at a rate equivalent to the endogenous glucose production rate. Blood glucose and plasma insulin concentrations were continuously monitored. Glucose turnover, glycolysis, and glycogen synthesis rates were determined by the 3H-glucose infusion technique. We showed that portal glucose infusion in RIPGLUT1 x GLUT24-/- mice did not induce the hypoglycemia observed in control mice but, in contrast, led to a transient hyperglycemic state followed by a return to normoglycemia; this glycemic pattern was similar to that observed in control Fe-mice and RIPGLUT1 x GLUT2-/- Fe-mice. Plasma insulin profiles during the infusion period were similar in control and RIPGLUT1 x GLUT2 /- Po- and Fe-mice. The lack of hypoglycemia development in RIPGLUT1 x GLUT2-/- mice was not due to the absence of GLUT2 in the liver. Indeed, reexpression by transgenesis of this transporter in hepatocytes did not restore the development of hypoglycemia after initiating portal vein glucose infusion. In the absence of GLUT2, glucose turnover increased in Po-mice to the same extent as that in RIPGLUT1 x GLUT2-/- or control Fe-mice. Finally, co-infusion of somatostatin with glucose prevented development of hypoglycemia in control Po-mice, but it did not affect the glycemia or insulinemia of RIPGLUT1 x GLUT2-/- Po-mice. Together, our data demonstrate that GLUT2 is required for the function of the hepatoportal glucose sensor and that somatostatin could inhibit the glucose signal by interfering with GLUT2-expressing sensing units. PMID- 11016448 TI - 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) inhibits insulin-stimulated glucose transport in 3T3-L1 adipocytes. AB - Incubation of skeletal muscle with 5-aminoimidazole-4carboxamide ribonucleoside (AICAR), a compound that activates 5'-AMP-activated protein kinase (AMPK), has been demonstrated to stimulate glucose transport and GLUT4 translocation to the plasma membrane. In this study, we characterized the AMPK cascade in 3T3-L1 adipocytes and the response of glucose transport to incubation with AICAR. Both isoforms of the catalytic alpha-subunit of AMPK are expressed in 3T3-L1 adipocytes, in which AICAR stimulated AMPK activity in a time- and dose-dependent fashion. AICAR stimulated 2-deoxy-D-glucose transport twofold and reduced insulin stimulated uptake to 62% of the control transport rate dose-dependently, closely correlating with the activation of AMPK. AICAR also inhibited insulin-stimulated GLUT4 translocation, assessed using the plasma membrane lawn assay. The effects of AICAR on insulin-stimulated glucose transport are not mediated by either adenosine receptors or nitric oxide synthase and are mediated downstream of phosphatidylinositol 3'-kinase stimulation. We propose that in contrast to skeletal muscle, in which AMPK stimulation promotes glucose transport to provide ATP as a fuel, AMPK stimulation inhibits insulin-stimulated glucose transport in adipocytes, inhibiting triacylglycerol synthesis, to conserve ATP under conditions of cellular stress. Investigation of the mode of action of AICAR and AMPK may, therefore, give insight into the mechanism of insulin action. PMID- 11016449 TI - Effect of cow's milk exposure and maternal type 1 diabetes on cellular and humoral immunization to dietary insulin in infants at genetic risk for type 1 diabetes. Finnish Trial to Reduce IDDM in the Genetically at Risk Study Group. AB - Type 1 diabetes is considered to be a T-cell-mediated autoimmune disease in which insulin-producing beta-cells are destroyed. Immunity to insulin has been suggested to be one of the primary autoimmune mechanisms leading to islet cell destruction. We have previously shown that the first immunization to insulin occurs by exposure to bovine insulin (BI) in cow's milk (CM) formula. In this study, we analyzed the development of insulin-specific T-cell responses by proliferation test, emergence of insulin-binding antibodies by enzyme immunoassay, and insulin autoantibodies by radioimmunoassay in relation to CM exposure and family history of type 1 diabetes in infants with a first-degree relative with type 1 diabetes and increased genetic risk for the disease. The infants were randomized to receive either an adapted CM-based formula or a hydrolyzed casein (HC)-based formula after breast-feeding for the first 6-8 months of life. At the age of 3 months, both cellular and humoral responses to BI were higher in infants exposed to CM formula than in infants fully breast-fed (P = 0.015 and P = 0.007). IgG antibodies to BI were higher in infants who received CM formula than in infants who received HC formula at 3 months of age (P = 0.01), but no difference in T-cell responses was seen between the groups. T-cell responses to BI at 9 months of age (P = 0.05) and to human insulin at 12 (P = 0.014) and 24 months of age (P = 0.009) as well as IgG antibodies to BI at 24 months of age (P = 0.05) were lower in children with a diabetic mother than in children with a diabetic father or a sibling, suggesting possible tolerization to insulin by maternal insulin therapy. The priming of insulin-specific humoral and T-cell immunity occurs in early infancy by dietary insulin, and this phenomenon is influenced by maternal type 1 diabetes. PMID- 11016450 TI - Anti-CD154 (CD40L) prevents recurrence of diabetes in islet isografts in the DR BB rat. AB - Islet transplantation for the treatment of autoimmune diabetes is more difficult because of the additional barrier presented by the autoimmunity. We tested the ability of hamster anti-rat CD154 to prevent recurrence of diabetes in renal subcapsular islet isografts in DR-BB (RT1uu) rats with established autoimmune diabetes. Experimental animals with established diabetes received intravenous injections of 15 mg/kg anti-CD154 on a specified schedule starting 2 days before renal subcapsular transplantation of an islet isograft. Control animals received either saline or hamster IgG. Plasma glucose levels >250 mg/dl over 3 days were used to indicate the recurrence of diabetes. Rats that received saline (n = 5) or control antibody (n = 3) had a recurrence of diabetes 6-11 days after transplantation. Histological examination of islet isografts from these rats showed complete destruction of the insulin-producing portion of the isograft with residual cells positive for glucagon. Recipient rats that received anti-CD154 at the 15-mg/kg dosage (n = 6) did not have a recurrence of diabetes for 308-461 days after transplantation. Islet isografts removed from the rats showed low levels of insulin immunoreactivity, high levels of insulin mRNA, and focal infiltration with lymphocytes but no evidence of islet destruction. Mean peak antibody concentration was 266 microg/ml and returned to undetectable levels by 67-88 days after transplantation. Rats that received anti-CD154 starting at 4-7 days after transplantation had a recurrence of diabetes within 11 days of the isotransplantation. Therefore, anti-CD154 as the sole immunomodulator prevented the recurrence of diabetes in islet isografts in rats with established autoimmune diabetes. This suggests that CD40/CD154 blockade is effective in preventing the insulitis or the effector phase of autoimmune diabetes. PMID- 11016451 TI - Adult human cytokeratin 19-positive cells reexpress insulin promoter factor 1 in vitro: further evidence for pluripotent pancreatic stem cells in humans. AB - Human pancreatic cells with a typical ductal phenotype and potential to proliferate can be obtained in vitro, but the differentiation capacity of these putative human pancreatic stem cells remains to be documented. We investigated the protein and mRNA expression of insulin promoter factor 1 (IPF-1) (or pancreas/duodenal homeobox 1), a transcription factor critical for pancreatic development and endocrine cell neogenesis, in human pancreatic ductal cells derived from cultured exocrine tissue. In vitro, exocrine cells rapidly adhered (within 12 h) and were de-/transdifferentiated to ductal cells after 3 days with a dramatic loss of amylase protein (n = 4, 92 +/- 3.3%, P < 0.05 vs. day 1) and a simultaneous increase of ductal cytokeratin 19 protein (n = 4, 3.4-fold on day 3 and 7-fold on day 9, P < 0.05 vs. day 1). IPF-1 protein and mRNA levels were low to undetectable in exocrine preparations before culture. After 2 days of culture, a 3.2-fold increase in IPF-1 protein was observed, corresponding to the characteristic 46-kDa protein in Western blots. Reverse transcriptase-polymerase chain reaction confirmed a 10.5-fold increase in IPF-1 mRNA levels after 3 days of culture (n = 5, P < 0.001 vs. day 1). Double immunocytochemistry showed direct evidence that IPF-1 appeared during culture in these exocrine-derived ductal cells (cytokeratin 7-positive) and was not merely in contaminating endocrine cells (chromogranin A-positive). In conclusion, we describe herein the first converging evidence on both the molecular and protein level that human cells with a typical ductal phenotype derived ex vivo from pancreatic exocrine tissue (obtained from healthy donors) can reexpress IPF-1 in culture, suggesting their pancreatic precursor/stem cell potential. PMID- 11016452 TI - Glucagon stimulates expression of the inducible cAMP early repressor and suppresses insulin gene expression in pancreatic beta-cells. AB - The hormone glucagon is secreted by the alpha-cells of the endocrine pancreas (islets of Langerhans) during fasting and is essential for the maintenance of blood glucose levels by stimulation of hepatic glucose output. Excessive production and secretion of glucagon by the alpha-cells of the islets is a common accompaniment to diabetes. The resulting hyperglucagonemia stimulates hepatic glucose production, thereby contributing to hyperglycemia of diabetes. The reduced insulin secretion in diabetes and resultant failure to suppress glucagon secretion by intra-islet paracrine mechanisms is believed to cause the hypersecretion of glucagon. Here, we report the discovery of a new mechanism by which glucagon suppresses insulin secretion. We show that glucagon, but not glucagon-like peptide 1 (GLP-1), or pituitary adenylyl cyclase-activating peptide (PACAP) specifically induces the expression of the transcriptional repressor inducible cAMP early repressor (ICER) in pancreatic beta-cells, resulting in a repression of the transcriptional expression of the insulin gene. Remarkably, glucagon, GLP-1, and PACAP all stimulate the formation of cAMP to a comparable extent in rat pancreatic islets, but only glucagon activates the expression of ICER and represses insulin gene transcription in beta-cells. These findings lead us to propose that hyperglucagonemia may additionally aggravate the diabetic phenotype via a suppression of insulin gene expression mediated by the transcriptional repressor ICER. PMID- 11016453 TI - Effect of dietary energy restriction on glucose production and substrate utilization in type 2 diabetes. AB - A total of 8 obese subjects with type 2 diabetes were studied while on a eucaloric diet and after reduced energy intake (25 and then 75% of requirements for 10 days each). Weight loss was 2, 3, and 3 kg after 5, 10, and 20 days, respectively; all of the weight lost was body fat. Fasting blood glucose (FBG) levels fell from 11.9 +/- 1.4 at baseline to 8.9 +/- 1.6, 7.9 +/- 1.4, and 8.8 +/ 1.3 mmol/l at days 5, 10, and 20, respectively (P < 0.05, baseline vs. 5, 10, and 20 days). Endogenous glucose production (EGP) was 22 +/- 2, 18 +/- 2, 17 +/- 2, and 22 +/- 2 pmol x kg(-1) lean body mass (LBM) x min(-1) (P < 0.05, days 5 and 10 vs. baseline). Gluconeogenesis measured by mass isotopomer distribution analysis provided 31 +/- 4, 41 +/- 5, 40 +/- 4, and 33 +/- 4%, respectively, of the EGP (NS); absolute glycogenolytic contribution to the EGP was 15 +/- 2, 11 +/ 2, 11 +/- 2, and 15 +/- 2 pmol x kg(-1) LBM x min(-1), respectively (P < 0.001, baseline vs. days 5 and 10 and day 10 vs. day 20). The blood glucose clearance rate increased significantly at day 20 (P < 0.05). Neither lipolysis nor flux of plasma nonesterified fatty acids were altered compared with baseline. In conclusion, severe energy restriction per se independent of major changes in body composition reduces both FBG concentration and EGP in type 2 diabetes, the reduction in EGP results entirely from a reduction of glycogenolytic input into blood glucose, and the duration of reduced glycogenolysis is short-lived after relaxation of energy restriction even without weight gain, but effects on plasma glucose clearance persist and partially maintain the improvement in fasting glycemia. PMID- 11016454 TI - Dexamethasone-induced insulin resistance in 3T3-L1 adipocytes is due to inhibition of glucose transport rather than insulin signal transduction. AB - Glucocorticoids reportedly induce insulin resistance. In this study, we investigated the mechanism of glucocorticoid-induced insulin resistance using 3T3 L1 adipocytes in which treatment with dexamethasone has been shown to impair the insulin-induced increase in glucose uptake. In 3T3-L1 adipocytes treated with dexamethasone, the GLUT1 protein expression level was decreased by 30%, which possibly caused decreased basal glucose uptake. On the other hand, dexamethasone treatment did not alter the amount of GLUT4 protein in total cell lysates but decreased the insulin-stimulated GLUT4 translocation to the plasma membrane, which possibly caused decreased insulin-stimulated glucose uptake. Dexamethasone did not alter tyrosine phosphorylation of insulin receptors, and it significantly decreased protein expression and tyrosine phosphorylation of insulin receptor substrate (IRS)-1. Interestingly, however, protein expression and tyrosine phosphorylation of IRS-2 were increased. To investigate whether the reduced IRS-1 content is involved in insulin resistance, IRS-1 was overexpressed in dexamethasone-treated 3T3-L1 adipocytes using an adenovirus transfection system. Despite protein expression and phosphorylation levels of IRS-1 being normalized, insulin-induced 2-deoxy-D-[3H]glucose uptake impaired by dexamethasone showed no significant improvement. Subsequently, we examined the effect of dexamethasone on the glucose uptake increase induced by overexpression of GLUT2-tagged p110alpha, constitutively active Akt (myristoylated Akt), oxidative stress (30 mU glucose oxidase for 2 h), 2 mmol/l 5-aminoimidazole-4-carboxamide ribonucleoside for 30 min, and osmotic shock (600 mmol/l sorbitol for 30 min). Dexamethasone treatment clearly inhibited the increases in glucose uptake produced by these agents. Thus, in conclusion, the GLUT1 decrease may be involved in the dexamethasone-induced decrease in basal glucose transport activity, and the mechanism of dexamethasone induced insulin resistance in glucose transport activity (rather than the inhibition of phosphatidylinositol 3-kinase activation resulting from a decreased IRS-1 content) is likely to underlie impaired glucose transporter regulation. PMID- 11016455 TI - Impaired capacity to lose visceral adipose tissue during weight reduction in obese postmenopausal women with the Trp64Arg beta3-adrenoceptor gene variant. AB - Controversy exists regarding the association between the Trp64Arg variant of the beta3-adrenoceptor gene and visceral obesity. The cross-sectional nature of most studies, the modest effect of the variant, and sex or ethnic differences between groups have contributed to discrepancies among investigations. To overcome these confounding factors, we examined the effect of the Trp64Arg variant on total and visceral adipose tissue loss, insulin sensitivity, and cardiovascular disease risk factors in response to weight reduction in obese older women. A total of 24 women (age 57 +/- 4 years), including 1 Trp64Arg homozygote, 10 Trp64Arg heterozygotes, and 13 normal homozygotes, were admitted to a weight reduction program of 13 +/- 3 months, with weight and nutritional intake stabilization established before testing. Total and regional adiposity were measured with dual energy X-ray absorptiometry and computed tomography, insulin sensitivity was measured by the hyperinsulinemic-euglycemic clamp technique, and a blood lipid profile was obtained. No baseline differences were noted in adiposity measurements, glucose disposal, and lipid profiles among carriers and noncarriers of the variant allele. In response to weight loss, carriers and noncarriers of the Trp64Arg allele had similar reductions in body weight (-16.4 +/- 5.0 vs. 14.1 +/- 6.2 kg, NS) and body fat (-10.0 +/- 5.2 vs. -11.5 +/- 3.9 kg, NS). However, loss of visceral adipose tissue was 43% lower in carriers of the Trp64Arg allele compared with noncarriers (-46 +/- 27 vs. -81 +/- 51 cm2, P = 0.05). Furthermore, there was less improvement in the total cholesterol-to-HDL cholesterol ratio (-0.18 +/- 0.54 vs. -0.72 +/- 0.56, P = 0.04) in carriers compared with noncarriers of the allele. Although glucose disposal improved in both groups, there was no difference in the magnitude of improvement between carriers and noncarriers of the variant allele. In conclusion, older obese women carrying the Trp64Arg beta3-adrenoceptor gene variant have an impaired capacity to lose visceral adipose tissue in response to prolonged caloric restriction. Despite these genetic differences in loss of intraabdominal adipose tissue, improvement in glucose disposal was similar between groups. PMID- 11016456 TI - Glycolaldehyde, a reactive intermediate for advanced glycation end products, plays an important role in the generation of an active ligand for the macrophage scavenger receptor. AB - Long-term incubation of proteins with glucose leads to the formation of advanced glycation end products (AGEs) that are recognized by AGE receptors. Glyoxal, glycolaldehyde (GA), and methylglyoxal are potential intermediates for the formation of AGE structures such as Nomega-(carboxymethyl)lysine (CML). We evaluated the contribution of these aldehydes to the formation of AGE structure(s), particularly the structure important for the receptor-mediated endocytic uptake of AGE proteins by macrophages. GA-modified bovine serum albumin (BSA), methylglyoxal-modified BSA (MG-BSA), and glyoxal-modified BSA (GO-BSA) were prepared, and their physicochemical, immunological, and biologic properties were compared with those of glucose-derived AGE-BSA. CML contents were high in GO BSA and low in GA-modified BSA (GA-BSA) but did not exist in MG-BSA. The fluorescence patterns of GA-BSA and MG-BSA were similar to those of glucose derived AGE-BSA but were weak in GO-BSA. Immunochemically, the antibody against non-CML structures of glucose-derived AGE-BSA reacted strongly with GA-BSA and weakly with GO-BSA but did not react with MG-BSA. The negative charge of these ligands increased to a similar extent. However, GA-BSA, but not MG-BSA or GO-BSA, underwent receptor-mediated endocytosis by the macrophage-derived cell line RAW 264.7, which was effectively inhibited by glucose-derived AGE-BSA, acetylated LDL, and oxidized LDL, which are well-known ligands for the macrophage type I and type II class A scavenger receptors (MSR-A). The endocytic uptake of GA-BSA by mouse peritoneal macrophages was also significant, but that by peritoneal macrophages from MSR-A-deficient mice was markedly reduced. Our results suggest that GA serves as an important intermediate for the generation of AGE structure(s) responsible for recognition by MSR-A. PMID- 11016457 TI - Activity of the glycosylating enzyme, core 2 GlcNAc (beta1,6) transferase, is higher in polymorphonuclear leukocytes from diabetic patients compared with age matched control subjects: relevance to capillary occlusion in diabetic retinopathy. AB - The exact mechanism for capillary occlusion in diabetic retinopathy is still unclear, but increased leukocyte-endothelial cell adhesion has been implicated. We examined the possibility that posttranslational modification of surface O glycans by increased activity of core 2 transferase (UDP-Glc:Galbeta1 3GalNAcalphaRbeta-N-acetylglucoaminyltr ansferase) is responsible for increased adhesion of leukocytes to vascular endothelium in diabetes. The mean activity of core 2 transferase in polymorphonuclear leukocytes isolated from type 1 and type 2 diabetic patients was higher compared with age-matched control subjects (1,638 +/- 91 [n = 42] vs. 249 +/- 35 pmol x h(-1) x mg(-1) protein [n = 24], P = 0.00013; 1,459 +/- 194 [n = 58] vs. 334 +/- 86 [n = 11], P = 0.01). As a group, diabetic patients with retinopathy had significantly higher mean activity of core 2 transferase compared with individuals with no retinopathy. There was a significant association between enzyme activity and severity of retinopathy in type 1 and type 2 diabetic patients. There was a strong correlation between activity of core 2 transferase and extent of leukocyte adhesion to cultured retinal capillary endothelial cells for diabetic patients but not for age-matched control subjects. Results from transfection experiments using human myelocytic cell line (U937) demonstrated a direct relationship between increased activity of core 2 transferase and increased binding to cultured endothelial cells. There was no relationship between activity of core 2 transferase and HbA(1c) (P = 0.8314), serum advanced glycation end product levels (P = 0.4159), age of the patient (P = 0.7896), and duration of diabetes (P = 0.3307). On the basis that branched O glycans formed by the action of core 2 transferase participate in leukocyte adhesion, the present data suggest the involvement of this enzyme in increased leukocyte-endothelial cell adhesion and the pathogenesis of capillary occlusion in diabetic retinopathy. PMID- 11016458 TI - Remodeling of networks of interstitial cells of Cajal in a murine model of diabetic gastroparesis. AB - Patients with long-standing diabetes commonly suffer from gastric neuromuscular dysfunction (gastropathy) causing symptoms ranging from postprandial bloating to recurrent vomiting. Autonomic neuropathy is generally believed to be responsible for diabetic gastropathy and the underlying impairments in gastric emptying (gastroparesis) and receptive relaxation, but the specific mechanisms have not been elucidated. Recently, it has been recognized that interstitial cells of Cajal generate electrical pacemaker activity and mediate motor neurotransmission in the stomach. Loss or defects in interstitial cells could contribute to the development of diabetic gastroparesis. Gastric motility was characterized in spontaneously diabetic NOD/LtJ mice by measuring gastric emptying and by monitoring spontaneous and induced electrical activity in circular smooth muscle cells. Interstitial cells of Cajal were studied by Kit immunofluorescence and transmission electron microscopy. Diabetic mice developed delayed gastric emptying, impaired electrical pacemaking, and reduced motor neurotransmission. Interstitial cells of Cajal were greatly reduced in the distal stomach, and the normally close associations between these cells and enteric nerve terminals were infrequent. Our observations suggest that damage to interstitial cells of Cajal may play a key role in the pathogenesis of diabetic gastropathy. PMID- 11016459 TI - Gene encoding the catalytic subunit p110beta of human phosphatidylinositol 3 kinase: cloning, genomic structure, and screening for variants in patients with type 2 diabetes. AB - Phosphatidylinositol (PI) 3-kinase is a key signaling molecule in insulin stimulated glucose transport. Therefore, we investigated the catalytic subunit p110beta, of human PI 3-kinase as a candidate gene for type 2 diabetes. Human p110beta gene was cloned from the placental genomic library. All 22 exons, intronic regions flanking the exons and 1.5 kb of the proximal/5' region of the p110beta gene, were screened for variants by single-strand conformation polymorphism analysis in 79 Finnish patients with type 2 diabetes . Allele frequencies of the variants were also determined in 77 nondiabetic control subjects. No variants were found in exons in diabetic patients. However, we identified two nucleotide polymorphisms in the proximal/5' region of the p110beta gene and a variation in the number of 2-bp repeat sequence (TA)n in intron 4. The allele frequencies did not differ between diabetic and control subjects. Our results may indicate that the catalytic subunit p110beta of PI 3-kinase plays such a fundamental role in the insulin-signaling pathway that structural variants are not likely to exist in that gene. The importance of the polymorphisms in the proximal/5' region of the p110beta gene for insulin signaling remains to be determined. PMID- 11016460 TI - NOD Idd5 locus controls insulitis and diabetes and overlaps the orthologous CTLA4/IDDM12 and NRAMP1 loci in humans. AB - A genome scan for B10-derived loci that reduce the frequency of diabetes and insulitis in NOD mice demonstrated a large region (34 cM) of linkage on the proximal end of chromosome 1. This locus was designated Idd5 and encompassed candidate genes including Il1r1, Il1r2, Stat1, Stat4, Nramp1, and Bcl2. In the current study, we have confirmed the existence of Idd5 by developing a series of congenic mouse strains that are resistant to diabetes and determined that Idd5 is actually two genes located within a 9.4-cM interval. Idd5.1 is in the proximal 1.5-cM portion of the interval and contains the candidates Casp8, Cflar (FLIP), Cd28, and Cd152 (CTLA4). Idd5.1 overlaps the orthologous CTLA4/IDDM12 locus in humans. Idd5.2 is in the distal 5.1-cM portion of the 9.4-cM interval and contains the candidates Nramp1, which has a functional polymorphism between NOD and B10, and Cmkar2 (CXCR2, interleukin [IL]-8 receptor alpha). Candidate genes eliminated by this analysis include Il1r1, Ilr2, Zap70, Orch5, Stat1, Stat4, Bcl2, Cmkar4 (CXCR4), and Il10. On its own, the Idd5 locus provides a significant amount of protection from diabetes (50% reduction from parental frequency) and when combined with another resistance locus (Idd3 on chromosome 3), provides nearly complete protection from diabetes and insulitis. PMID- 11016461 TI - The actions and interactions of nitric oxide in solid tumours. AB - The small molecule nitric oxide (NO) has undergone an image change since its identification as a biological messenger in 1987. It is a free radical, with a diverse range of actions in both physiological and pathological processes. Whilst over 30000 research papers have been written to date on NO, its role in tumour biology remains incompletely understood and research in this field is still in its infancy. NO would appear to have both tumour promoting and inhibiting effects which are presumed to be dependent on its local concentration within the tumour. Recently the relationships of NO to the tumour suppressor gene p53 have been experimentally elucidated, demonstrating how mutations of p53 may adversely affect the host by enhancing NO production. This review summarizes the brief history of this molecule, outlines its roles in the common solid tumours and suggests areas for future research. PMID- 11016462 TI - Radical surgery for gallbladder cancer: current options. AB - Gallbladder carcinoma is the most common malignancy of the biliary tract. There are still many controversies regarding the type of curative surgical treatment for each stage of the disease. The staging system used is the TNM classification of the International Union Against Cancer. Different patterns of spread characterize gallbladder cancer but the two main types are direct invasion and lymph node metastases; since only the depth of invasion can be easily recognized by imaging techniques, it becomes the main variable in choosing the appropriate surgical treatment. Most Tis and T1 tumours are incidentally discovered after cholecystectomy for cholelithiasis and no further therapy is requested; for pT1b tumours, relaparotomy with hepatic resection and N1 dissection is associated with a better survival. For T2 tumours, cholecystectomy with hepatic resection and dissection of N1-2 lymph nodes is the standard treatment, with a 5-year survival of 60-80%. The only chance of long-term survival for patients with a T3-T4 tumour is an extended operation combining an hepatic resection with an N1-2 dissection with or without excision of the common bile duct. A subset of patients with peripancreatic positive nodes or invasion of adjacent organs seems to benefit from a synchronous pancreaticoduodenectomy. PMID- 11016464 TI - One stop breast clinics--victims of their own success? A prospective audit of referrals to a specialist breast clinic. AB - AIMS: We aimed to assess the number of unnecessary referrals to a specialist breast clinic, with special reference to urgent referrals, and to compare referrals with published guidelines for referral to these clinics. METHODS: We carried out a prospective audit of new patient referrals from primary care to a specialist breast clinic. We assessed the total number of referrals, proportion of urgent and non-urgent referrals, proportion of unnecessary referrals according to published guidelines, waiting times for outpatient appointments and outcomes for these patients. RESULTS: Of the total of 321 referrals, 35% were urgent. Twenty-eight per cent of urgent referrals and 37% of non-urgent were inappropriate according to published guidelines. Ten per cent of referrals had breast cancer while 90% had benign disease or no pathology. CONCLUSIONS: There is a proliferation of guidelines (NHS, SIGN, BASO, Patients' Charter) for the organization of specialist breast clinics with the aim of providing rapid diagnosis for patients with malignant disease, or reassurance for symptomatic patients that they do not have breast cancer. However increasing numbers of patients are being referred to these clinics with minimal or no pathology, and this is not acknowledged by these guidelines. In this prospective study of referrals from primary care to a specialist breast clinic, one-third of referrals were inappropriate and this inevitably reduced the efficiency of the service provided for patients with significant symptoms. PMID- 11016463 TI - Ipsilateral local recurrence in relation to therapy and morphological characteristics in patients with ductal carcinoma in situ of the breast. AB - METHOD AND RESULTS: A standardized histopathological protocol has been designed, in which different histological characteristics of ductal carcinoma in situ (DCIS) are reported: nuclear grade (ng), growth pattern according to Andersen et al., necrosis, size of the lesion, resection margins and focality. Using this protocol a re-evaluation of a population-based consecutive series of 306 cases of DCIS has been done as well as a thorough clinical follow-up. After a median follow-up of 63 months, 13% have developed ipsilateral local recurrences, invasive and/or in situ. Ipsilateral local recurrence-free survival (IL-RFS) was significantly better for patients operated with mastectomy (ME) or breast conserving therapy (BCT) with radiotherapy (RT) than for patients operated with BCT without RT (5-year IL-RFS 96% vs 94% vs 79%, P<0.001). In the subgroup of BCT without RT there were significant differences in IL-RFS between histopathological subgroups: ng 1 + 2 (non-high grade) vs ng 3 (high grade; P=0.014), non-high grade without comedo-type necrosis vs non-high-grade with comedo-type necrosis vs high-grade (the Van Nuys classification system; P=0.025). Growth pattern (not diffuse vs diffuse) and margins (free vs involved or not evaluated) showed a tendency (P=0.07 and 0.05, respectively) to be associated to IL-RFS. In contrast, no significant differences in IL-RFS were found in subgroups based on mode of detection, focality or size. Ninety-four per cent of the local recurrences after BCT appeared at the previous operation site. CONCLUSIONS: In the BCT without RT group, combinations of either non-high grade and not a diffuse growth pattern or non-high grade and free margins identified groups (constituting approximately 30% of the patients) were at low risk of developing ipsilateral recurrences (6-10%), compared to a 31-37% recurrence risk in the remaining groups during the observed follow-up time. The beneficial effect of post-operative RT for these low-risk groups can be questioned, and should be studied further. PMID- 11016465 TI - Allelic loss in chromosomal region 1q21-23 in breast cancer is associated with peritumoral angiolymphatic invasion and extensive intraductal component. AB - AIMS: Breast cancer is the most frequent cancer in the female population and the involvement of chromosomal alterations is often implicated in the development of cancer. The aim of our study was to assess loss of heterozygosity (LOH) in chromosome 1 in relation to clinical and pathological parameters. METHODS: Tumours, corresponding normal tissues and peripheral blood samples from 50 women with operable breast cancer, were analysed by polymerase chain reaction (PCR) at 16 polymorphic DNA markers, on both the long and short arm of chromosome 1. RESULTS: There was a significant correlation between chromosomal region 1q21-23 and the presence of extensive intraductal component (EIC) and peritumoral angiolymphatic (PALI) invasion, both independent markers of local recurrence. CONCLUSIONS: Allelic loss in region 1q21-23 may be a valuable prognostic biological marker for the detection of local relapse in breast cancer, in combination with other histological and clinical parameters. PMID- 11016466 TI - Circum-areolar mastectomy with immediate reconstruction (CAMIR). AB - AIMS: To develop an oncologically safe and aesthetically acceptable technique for mastectomy using a myocutaneous flap and tissue expander for patients with neoplastic involvement of the nipple. METHODS: Fourteen consecutive patients (mean age 40 years) underwent a circum-areolar mastectomy with immediate reconstruction using a latissimus dorsi myocutaneous flap and tissue expander. Assessment of cosmesis was by review of pre- and post-operative photographs by an independent observer. RESULTS: During a mean follow-up of 11.4 months there have been no local recurrences. There were two deaths: pulmonary embolus (one) and distant metastatic disease (one). Assessment of cosmesis gave a score of 47 out of 56 (84%). CONCLUSIONS: In patients with tumours involving the nipple, mastectomy with immediate reconstruction may be used to achieve a good cosmetic result. The replacement nipple is fashioned from a circle of skin attached to the latissimus dorsi myocutaneous harvest. PMID- 11016467 TI - Prognosis and clinicopathological characteristics of Ib-IIb adenocarcinoma of the uterine cervix in patients who have had radical hysterectomy. AB - AIMS: The aim of the present study was to evaluate the outcome of patients with stage lb-IIb cervical adenocarcinoma treated with radical hysterectomy, and to determine the clinicopathological characteristics of those patients. METHODS: A total of 255 patients with cervical carcinoma stage Ib-IIb (57 adenocarcinoma and 198 squamous cell carcinoma) who had undergone radical hysterectomy were included in this study. Patient survival distribution was calculated using the Kaplan Meier method. RESULTS: The estimated 5-year survival rate for patients with adenocarcinoma was significantly poorer than that for patients with squamous cell carcinoma (77.9% vs 91.7%). The survival rate in stage Ib patients did not differ between two groups (95.8% vs 94.4% respectively). The incidence of lymph node involvement was significantly higher in patients with adenocarcinoma than in those with squamous cell carcinoma (31.6% vs 14.8%). Among patients receiving post-operative radiotherapy, the survival rate for adenocarcinoma (71.1%) was significantly poorer than that for squamous cell carcinoma (90.0%). When patients underwent radical hysterectomy, the survival rate for stage II patients with adenocarcinoma was significantly poorer than that for patients with squamous cell carcinoma. CONCLUSIONS: The higher incidence of lymph node involvement and lower response to post-operative radiotherapy are considered to be factors of poorer prognosis in cervical adenocarcinoma. PMID- 11016468 TI - Randomized trial of regional plus systemic fluorinated pyrimidine compared with systemic fluorinated pyrimidine in treatment of colorectal liver metastases. AB - AIM: We report a prospective randomized study comparing survival, response and toxicity in colorectal liver metastasis (CLM) patients treated by either hepatic arterial floxuridine (HAI) plus continuous systemic fluorouracil/folinic acid or systemic fluorouracil/folinic acid. METHODS: Eighty-four CLM patients received either HAI plus systemic fluorouracil/folinic acid or systemic fluorouracil/ folinic acid. RESULTS: Significantly more HAI plus systemic, compared with systemic only, patients developed WHO grade 3 or 4 diarrhoea (P=0.004), but significant quality of life differences were not detected. Liver metastasis partial response at 4 months after randomization was significantly greater (P=0.003) in HAI plus systemic (13/29, 45%) compared with systemic only (7/30, 23%) patients. There was no significant difference between groups in the proportion of patients who died from extrahepatic disease progression, or in survival. CONCLUSION: Combining regional with systemic fluorinated pyrimidines did not improve survival compared with systemic fluorinated pyrimidine. PMID- 11016469 TI - Elevated tumour marker CA19-9: clinical interpretation and influence of obstructive jaundice. AB - AIMS: The tumour marker CA19-9 has been promoted as a reliable test for the detection of pancreatobiliary malignancy, yet its diagnostic role remains poorly defined. In this study the clinical interpretation of a raised serum CA19-9 level has been evaluated, with particular reference to obstructive jaundice. METHODS: One hundred and sixty-four patients with a CA19-9 level above 33 U/ml were studied. Serum CA19-9 was compared with clinical diagnosis and correlated with serum bilirubin level. In a subgroup of jaundiced patients (16 benign and 15 malignant cases), follow-up CA19-9 levels were determined 2 weeks after biliary drainage. RESULTS: The median CA19-9 level was lower in benign cases (102 (IQR 50 264) U/ml) than those with pancreatobiliary tumours (910 (IQR 263-6170) U/ml; P<0.01), although the overlap was substantial. In benign jaundiced cases, a positive correlation was observed between bilirubin and CA19-9 elevation (R=0.41, P<0.01). Relief of jaundice was associated with a fall in CA19-9 level in all benign cases and in nine of the 15 with malignancy. CONCLUSION: Confident discrimination between benign and malignant disease could not be made on the basis of a solitary elevated CA19-9 measurement. Hyperbilirubinaemia was associated with a further deterioration in specificity and caution is warranted when interpreting the results in jaundiced patients. Overall, only one half of patients with an elevated CA19-9 level ultimately proved to harbour a malignancy. PMID- 11016470 TI - Prevention of implantation metastases after resection of proximal bile duct tumours with pre-operative low dose radiation therapy. AB - AIMS: Pre-operative endoscopic retrograde cholangiopancreatography (ERCP) with biliary drainage is associated with a greater risk of implantation metastases after resection of proximal bile duct tumours. In a previous study among patients who had undergone biliary drainage before resection, eight patients (20%) developed implantation metastases, within 1 year following resection. The aim of this analysis was to evaluate the results of pre-operative irradiation with regard to a possible reduction of implantation metastases. METHODS: Twenty-one patients with proximal bile duct tumours who had undergone resection following pre-operative irradiation were retrospectively analysed. Pre-operative radiation therapy consisted of three fractions of 3.5 Gy external beam irradiation of the hilar area. RESULTS: Pre-operative biliary drainage was performed in 19 patients (90%). All patients received pre-operative radiotherapy during which no complications were noted. None of the patients developed implantation metastases within a follow-up time of 2 to 79 months. CONCLUSION: The results of this study suggest that pre-operative radiotherapy in patients with a resectable proximal bile duct tumour who have undergone pre-operative drainage, decreases the risk of implantation metastases. To be certain about the role of pre-operative radiotherapy, a randomized study is required. Until then, we advocate standard low dose radiotherapy preceding resection in all patients with lesions suggestive of a proximal bile duct tumour who have undergone biliary drainage. PMID- 11016471 TI - Peritonectomy and hyperthermic antiblastic perfusion in the treatment of peritoneal carcinomatosis. AB - AIMS: Some low-grade malignant tumours arising in the abdomen tend to remain loco regionally confined to peritoneal surfaces, without systemic dissemination. In these cases complete surgical tumour cytoreduction followed by intra- or post operative regional chemotherapy has curative potential. The aim of this study was to evaluate the outcome for patients treated in this way. METHODS: Peritonectomy was performed, involving the complete removal of all the visceral and parietal peritoneum involved by disease. After peritonectomy, hyperthermic antiblastic perfusion was carried out throughout the abdominopelvic cavity for 90 min, at a temperature of 41.5-42.5 degrees C, with mitomycin C (3.3 mg/m2/l) and cisplatin (25 mg/m2/l) (for appendicular or colorectal primaries), or cisplatin alone (for ovarian primaries). Alternatively, the immediate post-operative regional chemotherapy was performed with 5-fluorouracil (13.5 mg/kg) and Lederfolin (125 mg/m2) (for colonic or appendicular tumours) or cisplatin (25 mg/m2) (for ovarian tumours), each day for 5 days. RESULTS: Thirty-five patients affected by extensive peritoneal carcinomatosis were submitted to peritonectomy, with no residual macroscopic disease in all cases except three. Twenty-six patients were able to undergo the combined treatment involving loco-regional chemotherapy. Complications were observed in 54% of the patients and led to death in four of them. At a mean follow-up of 17 months overall 2-year survival was 55.2%, with a median survival of 26 months. CONCLUSIONS: After a learning curve of 18 months the feasibility of the integrated treatment increased to more than 90%, while mortality decreased dramatically. The curative potential of the combined therapeutic approach seems high in selected patients with peritoneal carcinomatosis not responding to systemic chemotherapy. Careful selection of patients can minimize the surgical risk, but the treatment should currently be reserved for clinical trials. PMID- 11016472 TI - Prognostic significance of peri-operative blood transfusion following radical resection for oesophageal carcinoma. AB - INTRODUCTION: Peri-operative allogeneic blood transfusion may exert an immunomodulatory effect and has been associated with early recurrence and decreased survival following resection for several gastro-intestinal malignancies. The aim of this study was to evaluate the prognostic influence of transfusion requirements following radical oesophagectomy for cancer. METHODS: A consecutive series of 235 patients undergoing subtotal oesophagectomy with two field lymphadenectomy in a single centre from April 1990 to June 1999 were studied. RESULTS: The median age was 64 years (30-79) with a male to female ratio of 3:1. The predominant histological subtype was adenocarcinoma (n = 154) compared to squamous carcinoma (n = 81). To avoid the influence of surgical complications data were excluded from the 5.5% of patients suffering in-hospital mortality. In the remaining patients, median blood loss was 900 ml (200-5500) with 46% (103/222) requiring transfusion (median 3 units, range 2-21). Median survival of non-transfused patients was 36 months compared to only 19 months for those receiving transfusion (log-rank = 4.44; 1 df, P = 0.0352). Non-transfused patients had significantly higher 2 and 5-year survival rates of 62% and 41% respectively in contrast to only 40% and 25% in those receiving blood transfusion. Even after stratification of results according to disease stage or the presence of major complications, survival was significantly worse in those receiving transfusion. Multivariate analysis demonstrated that in addition to nodal status, > 4 units transfusion was an independent prognostic indicator. CONCLUSION: Post-operative transfusion is associated with a significantly worse prognosis following radical oesophagectomy. Meticulous haemostasis and avoidance of unnecessary transfusion may prove oncologically beneficial. PMID- 11016473 TI - Gastric cancer: which patients benefit from systematic lymphadenectomy? AB - AIMS: The purpose of this study was to evaluate the value of systematic lymphadenectomy (SLA) in curative resected gastric cancer patients with respect to long-term survival, peri-operative morbidity and mortality. METHODS: We reviewed our prospectively gathered database of 309 resected gastric cancer patients and analysed the outcome of 185 R0-resected patients (60%) with respect to peri-operative morbidity, mortality and long-term survival by comparing 81 patients resected with SLA (D2-group) versus 104 patients resected without SLA (D1-group). RESULTS: Overall 5-year survival rates of R0-resected patients (n = 173; exclusion of peri-operative mortality) amounted to 49% and did not differ significantly between D2- and D1-groups (53% vs 47%); P=0.344). Nevertheless, subgroups of patients taking a benefit from SLA could be defined. Gastric cancer patients without LN metastases (pTx pN0; n=78) and patients with LN metastases only in perigastric lymph nodes (pTx pN1; n=34) showed a significantly better long-term prognosis when SLA was performed (84% vs 51%; P=0.001). Regarding peri operative morbidity (38% vs 39%) and mortality (6% in each case) we could not find any differences between the D2- and D1-groups. CONCLUSIONS: We conclude that SLA is able to improve long-term survival for some tumour stages. Therefore SLA should be recommended as a standard procedure in all gastric cancer patients resected with curative intention. PMID- 11016474 TI - The role of telomerase in breast cancer. PMID- 11016475 TI - Unusual association between cutaneous melanoma and axillary metastasis from occult breast cancer detected by sentinel node biopsy. AB - We report a case of cutaneous Stage I melanoma associated with occult breast cancer detected incidentally during a sentinel node biopsy. A brief review of the literature is presented with particular emphasis on this association and on an examination of the theoretical link which may exist between melanoma and breast cancer. PMID- 11016476 TI - Papillary carcinoma-in-situ of the breast presenting as a fungating lesion. AB - We report a case of papillary breast carcinoma-in-situ which presented as a fungating lesion. To our knowledge this is the first such case to be reported in the English language literature. PMID- 11016477 TI - Cutaneous metastasis to the head and neck from colonic carcinoma. AB - A case of metastatic facial skin involvement from colonic carcinoma is reported where, despite radiotherapy given preoperatively, the tumour continued to grow. The patient remained disease free for 8 months after excision and reconstruction, but died of pulmonary metastases. PMID- 11016478 TI - Cholangiocellular carcinoma associated with segmental Caroli's disease. AB - This case report describes the association of cholangiocarcinoma arising in a liver segment with Caroli's disease. We demonstrate the diagnostic difficulties in recognizing the pathogenetic background and outline the indication for surgical treatment. PMID- 11016479 TI - Re: reflections and proposals for the standardization of lymphadenectomy for gastric cancer. PMID- 11016480 TI - Student assessment in undergraduate medical education in the United Kingdom, 1998. AB - This report describes the results of the project 'Assessment of Undergraduate Medical Education: Re-inventing the Wheel?' funded by the Nuffield Trust. The project was initiated in order to obtain information on current assessment practices in medical schools across the UK, to determine the extent of change as a consequence of the curricular reforms recommended by the General Medical Council (GMC) in 1993 and, if necessary, to stimulate debate about assessment and provide an impetus for change. The data obtained provide a detailed profile of the timing and nature of assessments used in medical schools and provide information for comparison with the survey of basic medical education in the British Isles carried out in 1977 and any subsequent studies. The study provides confirmatory and unexpected evidence. Schools are clearly revising their curricula consistent with recommendations made by the GMC. The main components of the study were as follows: a postal questionnaire sent to all UK medical schools, a national workshop, and four case studies of innovative assessment practices. PMID- 11016481 TI - Ischemic cardiomyopathy and the cellular renin-angiotensin system. AB - BACKGROUND: Ischemic cardiomyopathy produced by non-occlusive coronary artery constriction is characterized by left ventricular failure and right ventricular dysfunction, but whether the local renin-angiotensin system (RAS) is implicated in myocyte dysfunction and cell death remains unclear. METHODS: Changes in single cell mechanics, the localization of the various constituents of RAS in the myocardium, and the effects of angiotensin II (Ang II) stimulation on myocyte performance and cell death were measured. RESULTS: Chronic ischemia is coupled with alterations in the mechanical properties and calcium (Ca2+) transients of the remaining viable myocytes. The abnormalities in myocyte mechanics consist of depression in peak shortening and velocity of shortening. Moreover, peak systolic Ca2+ is significantly decreased in the cells. In vitro stimulation with Ang II ameliorates myocyte function and systolic Ca2+. Additionally, adult myocytes express genes for renin, angiotensinogen, angiotensin-converting enzyme (ACE), and Ang II receptors. Renin, ACE, and Ang II receptors mRNAs increase under the setting of impaired coronary perfusion. Similarly, the percentage of myocytes containing renin, Ang I, and Ang II increases as well. In vitro studies of neonatal and adult ventricular myocytes indicate that Ang II triggers programmed myocyte cell death and this phenomenon is mediated by activation of the AT1 receptor sub-type. Importantly, the AT1-receptor blocker, losartan, completely inhibits apoptosis. CONCLUSIONS: These multiple observations are consistent with the notion that Ang II may exert 3 separate functions on the heart: (1) stimulation of myocyte hypertrophy, (2) amelioration of myocyte contractile performance, and (3) activation of the suicide program of myocytes. PMID- 11016482 TI - Behavioral and environmental factors contributing to the development and progression of congestive heart failure. PMID- 11016483 TI - Cardiovascular and peripheral rehabilitation. PMID- 11016484 TI - Sudden cardiac death syndrome and pump dysfunction: the link. AB - Clearly, sudden cardiac death syndrome in heart failure is linked to severely perturbed neurohumoral, hemodynamic, and mechanical systems. Routine antiarrhythmic drug therapy has not proven beneficial and, therefore, there is no justification for using these agents in unselected heart failure patients who are without significant symptomatic ventricular arrhythmia. Aggressive treatment of the failure syndrome seems most important. Because many of the problematic arrhythmias arise from triggering automaticity, which is known to occur in excessive ventricular stretch and wall stress, systemic vascular "unloading" with vasodilators and angiotensin-converting enzyme inhibitors is likely helpful. The most recent American College of Cardiology/American Heart Association Guidelines regarding therapeutic management of heart failure suggest that aggressive pharmacologic treatment of asymptomatic ventricular arrhythmias is best avoided. To be considered strongly for pharmacologic prescription or for implantation of a tachyarrhythmia termination device, a patient should have symptomatic ventricular tachycardia with an episode of syncope or sudden cardiac death syndrome rather than simply having palpitations of asymptomatic, unsustained ventricular tachycardia. Indeed, aggressively treating congestive heart failure with medication often eliminates potentially life-threatening arrhythmias. Appropriate use of vasodilators and, particularly, angiotensin-converting enzyme inhibitors is important. Correction of fluid balance and electrolyte disorders may be helpful to address symptoms and certainly is likely to decrease the potential for morbidity and mortality. On occasion it may be necessary to consider bradyarrhythmia pacemaker insertion or the use of atrioventricular nodal-ablation techniques with subsequent ventricular or atrioventricular pacer insertion. Obviously, sudden cardiac death due to sudden heart block or asystole might be attenuated with this strategy. PMID- 11016485 TI - Sudden death in heart failure patients: effects of optimized medical therapy. AB - Although medical therapy, particularly with angiotensin-converting enzyme (ACE) inhibitors, has been demonstrated to prolong life in patients with chronic heart failure, the effect of standard medical therapy on sudden unexpected death in patients with heart failure is less well understood. Recent clinical trials have provided new insights into this growing problem. The impact of modern medical therapy for heart failure, including ACE inhibitors, beta-adrenergic antagonists, digoxin, calcium channel antagonists, and antiarrhythmic interventions will be discussed. PMID- 11016486 TI - Cardiac transplantation in patients with refractory ventricular arrhythmias. PMID- 11016487 TI - Heart donor management and expansion of current donor selection criteria. PMID- 11016488 TI - Outpatient parenteral inotropic therapy for advanced heart failure. AB - BACKGROUND: Patients with advanced heart failure generally have hemodynamic perturbation characterized by low cardiac output and high ventricular filling pressures. This creates a clinical milieu with profound symptomatology that includes weakness, fatigue, and fluid-retention states causing peripheral edema, mesenteric congestion, and dyspnea syndromes. Great morbidity including hospital admissions and readmissions as well as high mortality rates ensue. Though medication and/or surgical intervention often attenuate heart failure symptomatology, morbidity, and mortality, some patients reach more advanced stages despite aggressive maneuvers. Indeed, patients presenting with acute decompensation of chronic congestive heart failure frequently receive parenteral inotropic drugs during their hospitalization with clinical improvement. Because these agents generally increase cardiac output and reduce pre-load and afterload, they can be lifesaving. Some patients, however, have symptomatic and hemodynamic rebound to worsened heart failure states during or shortly after inotrope weaning. METHODS: It was, then, a logical step to segue from acute inpatient inotrope infusion to long-term administration of these drugs in the outpatient setting when patients were dependent on these agents. Dopamine, dobutamine, and milrinone are all generally available inotropes that have been used singly or in combination in a chronic outpatient infusion setting. CONCLUSIONS: Data from a few small clinical trials and anecdotal case experience suggest that these drugs result in both hemodynamic and clinical improvement that is generally sustained during chronic administration, and even noted long after discontinuation of infusions in some patients. Some reports have suggested that intermittent infusion therapy in outpatients (so-called pulsed therapy) is effective in attenuating congestive heart failure symptoms long term, with more data supporting chronic infusion of these agents. Though questions regarding safety of these agents have been raised, a reasonable compendium of data published to date supports the contention that inotropic drugs used in this fashion ameliorate symptoms. Legitimate concern may be raised regarding exacerbation of arrhythmias with subsequent sudden cardiac death syndrome; however, in severely symptomatic heart failure patients, the trade-off between symptomatic amelioration and the chance of sudden cardiac death may be worthwhile. Unfortunately, precise guidance regarding the best drug, dose, optimal administration technique, weaning protocol, and actual risk/benefit ratio are not well characterized. Practice as been guided, in large part, by anecdotal experience. However, it appears that chronic or pulsed outpatient parenteral inotropic infusion therapy is frequently prescribed and that this treatment option is an effective alternative for carefully selected patients with severely symptomatic and advanced heart failure. Formulating optimal protocols for home inotropic drug infusion therapy by conducting properly designed clinical trials will be an essential endeavor. PMID- 11016489 TI - The case against outpatient parenteral inotropic therapy for advanced heart failure. PMID- 11016490 TI - Left ventricular reduction surgery with mitral valve repair. AB - The echocardiographic changes and functional outcome from mitral valve repair, combined with partial left ventriculectomy (PLV), the Batista procedure, were investigated. From May 1996 to August 1997 the operation was performed on 57 patients, primarily (95%) transplant candidates with idiopathic dilated cardiomyopathy. All had been Class IV (36.8% had improved to Class III by the time of surgery) on medical therapy, including 40% hospitalized on inotropes and 3 patients on intra-aortic balloon pumps. The mean cardiac index was 2.1+/-0.6 L/min/m2, with a wedge pressure of 24+/-8 mm Hg, and norepinephrine levels of 855+/-500 pg/mL (normal 80-526 pg/mL). There were 2 in-hospital mortalities (3.5%). At 3 months there were significant persistent changes in LV end-diastolic diameter (8.1+/-1.0 cm-6.3+/-0.9 cm) and ejection fraction (13.6+/-6%-23+/-7.7%). Subjective improvement included a mean change in NYHA functional class from 3.7 to 2.2, and objective changes included improvement in peak oxygen consumption from 10.6+/-4 mL/kg/min to 15.4+/-4.5 mL/kg/min. Actuarial survival at 1 year was 82.1%; and freedom from death, relisting for transplantation, and need for LVAD support was 58%. The earliest "learning curve" experience with this relatively unstudied operation warrants careful investigation. Many questions remain to be answered, but many patients have demonstrated early subjective and objective improvement. PMID- 11016491 TI - Dynamic cardiomyoplasty: expectations and results. AB - BACKGROUND: Cardiac transplantation is an option for only a small minority of patients with severe, chronic congestive heart failure. Transformed skeletal muscle has the potential to provide a durable form of fatigue-resistant muscle power to assist the cardiovascular system. METHODS: Since the first dynamic cardiomyoplasty performed by Carpentier and colleagues in 1985, the techniques to make this operative procedure an effective assist to the failed myocardium have been refined. Much has been learned about optimum patient selection so that perioperative morbidity and mortality is minimized. The cardiomyostimulator developed by Medtronic has undergone several revisions, primarily so that the stimulation of the muscle wrap is adaptive to variations in cardiac performance, such as those that occur with arrhythmia or during exercise. RESULTS: A review of the data to date suggest a substantial number of patients who undergo the procedure experience an enhanced quality of life, and an alleviation of their heart failure symptoms. Details of patient selection underscore that cardiomyoplasty is not an alternative to transplant. Seriously compromised patients with low peak oxygen uptake and very low ejection fractions have the highest short term mortality. CONCLUSIONS: Dynamic cardiomyoplasty remains a promising surgical option for patients with moderately-severe congestive heart failure (i.e. NYHA Class III patients). The ongoing randomized trial comparing cardiomyoplasty to standard medical therapy is expected to provide more detailed information about the benefits of this procedure. PMID- 11016492 TI - Mitral valve repair in cardiomyopathy. PMID- 11016493 TI - Selection and management of ventricular assist device patients: the Muenster experience. AB - BACKGROUND: Because of the growing discrepancy between the availability of donor organs and the number of patients with end-stage heart disease who need heart transplantation, a larger proportion of patients waiting for a suitable donor heart require pre-operative mechanical circulatory assistance. The criteria for the selection and management of these patients as applied at Muenster University Hospital are reviewed. METHODS: The study population consists of 631 patients referred to our center for transplantation between January 1, 1990, and December 31, 1996. Two hundred ninety-seven patients were listed for transplantation and 157 were transplanted. Of 41 patients who underwent implantation of a ventricular assist device (n = 34, Novacor; n = 6, TCI HeartMate; n = 1, Medos), 39 received the device as a bridge to transplantation and 2 as permanent support. For the purpose of the analysis, the study population was divided into 3 groups (elective bridging, urgent bridging, emergency bridging) and compared with heart transplant candidates who did not require mechanical circulatory assistance. RESULTS: Patients who underwent elective or urgent assist-device implantation were younger and had greater hemodynamic compromise than the remainder of patients waiting for heart transplantation, as suggested by a higher functional class and lower mean arterial pressure, cardiac index, serum sodium, and pulmonary artery wedge pressure. Survival of patients who electively underwent implantation of an assist device was better than that of patients who were stable on the waiting list and did not undergo heart transplantation during follow-up. CONCLUSIONS: This finding suggests that earlier implantation of assist devices may facilitate resolution of organ dysfunction before heart transplantation. PMID- 11016494 TI - Determination of candidacy for mechanical circulatory support: importance of clinical indices. PMID- 11016495 TI - Left ventricular assist device implantation: short and long-term surgical complications. AB - Long-term implanted left ventricular assist devices (LVADs) have significantly improved the care of patients awaiting heart transplantation and will provide an alternative therapy to select patients with heart failure. However, although the technology and clinical results continue to improve, LVAD implantation is still associated with a significant level of complications. Left ventricular assist device-associated complications can be broadly divided by their temporal occurrence. Early complications include perioperative hemorrhage, air embolism, and right ventricular failure. Beyond the perioperative period, late complications consist primarily of infection, thromboembolism, and primary device failure. An improved understanding of the mechanisms involved should aid the clinician in further reducing the incidence of these occurrences. PMID- 11016496 TI - Implantable LVAD insertion in patients with previous heart surgery. AB - Implantable LVAD insertion and support poses technical obstacles in patients with previous heart operations. In this study of 135 LVAD patients (64 pneumatic HeartMate; 39 vented-electric HeartMate; 32 Novacor devices), 72 (53%) had previous heart surgery. For patients with previous coronary artery bypass, the grafts were managed by replacement or preservation. For patients with ventricular aneursym (or aneursmysectomy), or partical left ventriculectomy, apical cannulation had to be modified to maintain precise unobstructed orientation of the inflow cannula towards the mitral valve. Mechanical mitral and aortic prostheses were rereplaced with tissue valves. Implantable cardioverter defibrillators were removed, or the LVAD pump was placed intraperitoneal and the generator was left in the abdominal wall pocket. There were no significant differences in reoperations for bleeding (22% primary vs 23% reoperations), perioperative RVAD support (12% primary vs 7% reoperation) or survival to transplant (82% primary vs 72% reoperations). In conclusion adjustment of standard LVAD implant technique can successfully manage most problems posed by patients with previous heart surgery. PMID- 11016497 TI - Emergency mechanical circulatory support: still viable? PMID- 11016498 TI - Demographic scope and economic magnitude of contemporary organ replacement therapies. AB - This analysis draws upon a variety of sources to provide a tally of the number of patients receiving organ replacement therapies and the costs associated with the provision of such therapies. Constituent data were available from treatment specific patient registries, peer reviewed reports in scientific literature, business publications, and industry sources. The magnitude and economic scope of the contemporary organ replacement enterprise were found to be much larger than is generally recognized. In the year 2000, the lives of over 20 million patients will be sustained, supported, or significantly improved by functional organ replacement. The impacted population grows at over 10% per year. Worldwide, first year and follow-up costs of organ prosthesis exceeds $300 billion US dollars per year and represents between 7 and 8% of total worldwide health care spending. Remarkably, in the United States, the costs of therapies enabled by organ replacement technology exceed 1% of the Gross Domestic Product. These findings constitute an incontestable tribute to the scientific significance and medical impact of the still nascent field of substitutive medicine. At the same time, the enormous magnitude of resources dedicated to organ replacement raises several issues related to overall cost effectiveness of current modalities and raises challenges and opportunities for future technical developments. PMID- 11016499 TI - New type of matrix support for bone marrow cell cultures: in vitro culture and in vivo transplantation experiments. AB - A new type of bone marrow cell culture system was developed by using a highly porous substrate matrix, i.e., porous polyvinyl formal (PVF) resin. Murine bone marrow (BM) cells were cultured without the use of exogenous growth factors in a three-dimensional matrix support made of collagen coated porous PVF resin. To examine the optimal conditions for highest stromal cell density, short-term and long-term in vitro culture experiments using PVF were performed. In the short term culture experiments, it was found that cubes of PVF (10 x 10 x 2 mm and 130 microm in pore size) coated with type I collagen with a seeding density of 2x10(7) BM cells offered the most appropriate culture conditions. In the long term cultures, BM cells in PVF maintained their viability for up to 6 weeks. In another series of re-inoculation experiments, freshly isolated BM cells were inoculated onto the already developed stromal layer. In this study, a higher cell density of the stromal layer was obtained in the PVF culture compared with those in the control dish culture. Based upon the results of in vitro experiments, in vivo transplantation studies were also performed. Histologic examinations of the subcutaneously transplanted PVF with stroma revealed host derived hematopoiesis inside the PVF matrix. Moreover, survival of approximately 15% of the transplanted BM cells that were cultured in PVF were confirmed in X-ray irradiated recipients. From these results, it is suggested that PVF resin is a promising three-dimensional substrate for BM cell culture and that it can maintain hematopoietic stem cells or progenitor cells after transplantation. PMID- 11016500 TI - An investigation of blood flow behavior and hemolysis in artificial organs. AB - In our previous study, in vitro hemolysis tests showed that collision flow against wall roughness had an effect on hemolysis when the flow velocity was more than 3 m/s and surface roughness was more than Ra = 1.54 microm. However, the specific portion of the flow on the wall that induced hemolysis was not clarified. Therefore, the purpose of this study was to present the relationship between flow behavior and hemolysis by means of in vitro tests and computational fluid dynamics (CFD) analysis. We investigated the relationship between the location of surface roughness and hemolysis. In CFD, we investigated the flow behavior on the wall. The highest rate of hemolysis was observed in a region around the center of the surface roughness on the bottom plate. On CFD analyses, the flow behavior included the highest wall shear stress (304 Pa) and the highest flow acceleration (2.8 m/s2) around the center of the bottom plate. Therefore, it is concluded that the causes of hemolysis during collision flow depend upon wall shear stress and flow acceleration. PMID- 11016501 TI - Hydrodynamic characteristics of artificial lungs. AB - An artificial lung is used during cardiopulmonary bypass to oxygenate blood and to control blood temperature. The pressure drop-flow rate characteristics of the membrane compartment in three hollow fiber membrane oxygenators were determined in vitro to characterize design features. Results are presented in a unique dimensionless relationship between Euler number, N(Eu) (ratio of pressure drop to kinetic energy), and Reynolds number, N(Re) (ratio of inertial to viscous forces), and are a function of the device porosity, epsilon, and a characteristic device length, xi, defined as the ratio of the mean blood path and manifold length: [equation in text]. This dimensionless approach allows us (1) to compare oxygenators independently, and (2) to relate water tests to blood. PMID- 11016502 TI - A new tracheal bioartificial organ: evaluation of a tracheal allograft with minimal antigenicity after treatment by detergent. AB - The aim of this study was to reduce the antigenicity of tracheal allografts by detergent treatment. We attempted to apply these grafts to tracheal immunosuppressant-free allotransplantation. Fresh tracheal grafts were harvested from donor beagle dogs and treated with a detergent at 4 degrees C for 48 hours. By using treated grafts, we have performed tracheal immunosuppressant-free transplantation in six dogs at an intrathoracic five-ring defect. Implanted grafts were covered with an omental pedicle. In five of the six grafts, complete removal of the epithelium and mixed glands was recognized with both light microscopy and scanning electron microscopy. The appearance of the cartilage cells in the grafts was similar to those in fresh trachea. Five dogs that received detergent treated grafts survived uneventfully. The grafts had been incorporated by the host trachea without stenosis. On the one tracheal graft in which removal of the epithelium was incomplete (noted after implantation), moderate stenosis occurred 1 month after placement. These results suggest that removal of the tracheal epithelium and mixed glands can remarkably reduce tracheal antigenicity. A tracheal graft that has its epithelium and mixed glands removed can be used in tracheal immunosuppressant-free allotransplantation. PMID- 11016503 TI - Understanding spontaneous output fluctuations of an amperometric glucose sensor: effect of inhalation anesthesia and use of a nonenzyme containing electrode. AB - Implantable glucose sensors are often unstable in vivo. Possible causes include local oscillations of glucose or oxygen levels, fluctuation of interferants, and external electromagnetic interference. To better understand glucose versus nonglucose mediated fluctuations, we compared sensors fabricated with glucose oxidase versus blank electrodes without enzyme in rabbits. We also investigated the effect of general anesthesia. We used power spectral analysis to investigate transmitted signals from amperometric peroxide sensing devices 2-3 weeks after subcutaneous implantation. Fasted animals were studied for 90 minutes in the conscious state and for 90 minutes during halothane anesthesia. Animals exhibited almost no body movement during the studies. In the conscious state, enzyme active sensors demonstrated more oscillations than blank electrodes at almost all frequencies from 2 to > 8 cycles per hour. This finding suggested that the spontaneous fluctuations were secondary to local changes in glucose or oxygen. Because fluctuations were not seen in the blank electrode, periodic changes in interferant concentrations, electromyographic activity, or in external electromagnetic interference are unlikely. General inhalation anesthesia was associated with markedly reduced sensor output fluctuation at almost all frequencies in enzyme active sensors. We conclude that fluctuation of electrochemical glucose sensor output, unrelated to fluctuations in blood glucose, is likely secondary to spontaneous changes in the local concentration or vascular delivery of glucose or oxygen. Anesthesia may have stabilized blood flow, preventing normal spontaneous autoregulatory variation. PMID- 11016505 TI - Right ventricular outflow tract reconstruction using a Goretex membrane monocusp valve in infant animals. AB - We examined the feasibility of using a polytetrafluoroethylene membrane (goretex) valve and transannular patch (TAP) for right ventricular outflow tract (RVOT) reconstruction in growing animal models. Eleven infant goats (Group A) and 12 infant sheep (Group B) underwent RVOT reconstruction under cardiopulmonary bypass. In Group A, a monocusp valve was constructed of goretex, and the RVOT was roofed over utilizing a TAP of bovine pericardium. In Group B, both a monocusp valve and a TAP were constructed of goretex. Animals were sacrificed at 6 or 12 postoperative months. Two goats in Group A died at 9 days and 4 months postoperatively due to RVOT obstruction caused by thrombus formation. Seven goats in Group A and 11 in Group B showed no pressure gradient across the valve. All animals in Group B revealed mild to moderate valvular insufficiency, but no ventricular deterioration. In contrast to the bovine pericardium, which showed a prominent fibroinflammatory reaction with calcified areas causing TAP shrinkage and RVOT stenosis, there was minimal calcification and inflammatory reaction directed against the goretex valve and TAP. We conclude that goretex can be used as a material from which to create both a monocusp valve and TAP for long-term RVOT reconstruction. PMID- 11016504 TI - Cardiac function after eight hour storage by using polyethylene glycol hemoglobin versus crystalloid perfusion. AB - Efforts to extend myocardial preservation for transplantation by crystalloid perfusion have been limited by edema and compromised function. We hypothesized that hypothermic perfusion preservation with a polyethylene glycol (PEG) conjugated hemoglobin solution may extend preservation times. The purpose of this study was to compare cardiac function after continuous perfusion by using a hypocalcemic, normokalemic crystalloid perfusate with and without the addition of PEG-hemoglobin (Hb). The hearts of 20 anesthetized and ventilated New Zealand White rabbits were harvested after cold cardioplegic arrest. Group I (n = 10) hearts were continuously perfused with a hypocalcemic, normokalemic 3% bovine PEG Hb solution at 20 degrees C and 30 mm Hg for 8 hours. Group II (n = 10) hearts were continuously perfused with an identical crystalloid solution without PEG-Hb for 8 hours under the same conditions as group I hearts. Cardiac function was measured with a left ventricular force transducer after transfer to a standard crystalloid Langendorff circuit at 37 degrees C and an aortic root pressure of 59 mm Hg. After 8 hours of perfusion preservation, heart rate was similar for groups I and II (p = not significant [NS]). Coronary blood flow after and during preservation was similar between PEG-Hb and crystalloid preserved hearts (p = NS). Left ventricular developed pressure, peak dP/dt, and peak -dP/dt were superior in hearts preserved with PEG-Hb. Percent water of total ventricular weight was 82.0% for group I and 81.6% for group II (p = NS). Continuous perfusion preservation of rabbit hearts for 8 hours with a hypocalcemic normokalemic PEG-Hb based solution at 30 mm Hg and 20 degrees C yields left ventricular function that is superior to perfusion with a similar crystalloid solution without PEG-Hb, despite similar myocardial edema and coronary flow. Extended cardiac perfusion preservation with this PEG-Hb based solution deserves further study, including comparison with traditional cardioplegic preservation solutions. PMID- 11016506 TI - Dynamic cardiomyoplasty decreases myocardial workload as assessed by tissue tagged MRI. AB - The effects of dynamic cardiomyoplasty (CMP) on global and regional left ventricular (LV) function in end-stage heart failure still remain unclear. MRI with tissue-tagging is a novel tool for studying intramyocardial motion and mechanics. To date, no studies have attempted to use MRI to simultaneously study global and regional cardiac function in a model of CMP. In this study, we used MRI with tissue-tagging and a custom designed MR compatible muscle stimulating/pressure monitoring system to assess long axis regional strain and displacement variations, as well as changes in global LV function in a model of dynamic cardiomyoplasty. Three dogs underwent rapid ventricular pacing (RVP; 215 BPM) for 10 weeks; after 4 weeks of RVP, a left posterior CMP was performed. After 1 year of dynamic muscle stimulation, the dogs were imaged in a 1.5 T clinical MR scanner. Unstimulated and muscle stimulated tagged long axis images were acquired. Quantitative 2-D regional image analysis was performed by dividing the hearts into three regions: apical, septal, and lateral. Maximum and minimum principal strains (lambda, and lambda2) and displacement (D) were determined and pooled for each region. MR LV pressure-volume (PV) loops were also generated. Muscle stimulation produced a leftward shift of the PV loops in two of the three dogs, and an increase in the peak LV pressure, while stroke volume remained unchanged. With stimulation, lambda1 decreased significantly (p<0.05) in the lateral region, whereas lambda2 increased significantly (p<0.05) in both the lateral and apical regions, indicating a decrease in strain resulting from stimulation. D only increased significantly (p<0.05) in the apical region. The decrease in strain between unassisted and assisted states indicates the heart is performing less work, while maintaining stroke volume and increasing peak LV pressure. These findings demonstrate that the muscle wrap functions as an active assist, decreasing the workload of the heart, while preserving total pump performance. PMID- 11016507 TI - Can a linear electrical analog model of a mechanical valve predict flow by using a pressure gradient? AB - The objective was to determine whether a previously developed technique for biological aortic valves could predict flow through a mechanical valve. An electrical analog model of the aortic valve that includes compliance, resistance, and inertance parameters, and corresponding second order differential equations was used to predict flow given a pressure gradient, as previously reported. Simulated pressures and flow were recorded by using a pulse duplicator system. The heart rate was varied from 60 to 180 bpm, and the stroke volume was varied from 22 to 67 cc. Resistance, inertance, and compliance parameters of the governing differential equation were estimated by using a least-squares fit to the measured flow at 120 bpm and 50 cc stroke volume. By using these parameter estimates, flow was calculated for other heart rates and stroke volumes. To achieve a better flow prediction, a nonlinear filter (third order polynomial range calibration equation) was applied to the output of the linear model (flow). The mean error, full-scale error, and spectral error in magnitude and phase between measured and predicted flow were compared. Error in mean flow ranged from 3% at medium flow rates to 90% at low flow rates. The maximum and minimum full scale errors were 12% and 5%, respectively. Error in the harmonics of measured and calculated flow ranged from 0% to 55%. Larger errors were usually present at the higher harmonics. The agreement between measured and calculated flow was better at normal and high flows but rather poor at low flows. The nonlinear filter (range calibration equation) was unable to account for the discrepancies between the measured and calculated flow over all flow ranges. It seems that this linear model and nonlinear filter have limited application, and an alternate nonlinear approach may produce better results. PMID- 11016508 TI - Survival outcomes of patients with giant cell myocarditis bridged by ventricular assist devices. AB - Giant cell myocarditis is a highly lethal disorder characterized by rapidly progressive congestive heart failure. The aim of this study was to describe the clinical course of patients with giant cell myocarditis who received a ventricular assist device. Patients with giant cell myocarditis were identified from the Multicenter Giant cell Myocarditis Registry. Bridging to cardiac transplantation in the giant cell myocarditis patients who received a ventricular assist device was compared with bridging in the general population of heart failure patients, as reported in the literature. Median posttransplantation survival for patients with giant cell myocarditis who received and did not receive ventricular assist devices was calculated by the Kaplan-Meier method and compared with use of the log-rank test. Nine patients with giant cell myocarditis who received ventricular assist devices were identified. Seven patients survived to transplantation, four were alive 30 days posttransplantation, and two survived to 1 year. The rate of successful bridging to transplantation in seven of nine patients (78%) is similar to that reported for other ventricular assist device recipients. Posttransplantation survival of 57% (4 of 7) at 30 days and 29% (2 of 7) at 1 year was significantly lower compared with 93% 1-year survival of the 30 patients with giant cell myocarditis who did not receive ventricular assist devices before transplantation (p<0.001). Ventricular assist devices can be an effective bridge to transplantation for patients with heart failure caused by giant cell myocarditis. Although their posttransplantation survival was poor in our series, a few patients had long-term survival. PMID- 11016509 TI - Emergency implantation of a left ventricular assist device in adolescents with biventricular failure. AB - Adolescents with congestive cardiomyopathy who present with intractable arrhythmia or progressive ventricular failure have a very poor prognosis and often die awaiting cardiac transplantation (CTx). We present our recent experience with a pneumatically powered left ventricular assist device (LVAD) implanted emergently to salvage adolescents with severe biventricular failure. Four patients, aged 15-17 years, body surface areas of 1.5-1.7 m2, with dilated cardiomyopathy (LV diastolic dimension, 7.1-8.3 cm); two presented with cardiovascular collapse, one with refractory ventricular tachycardia, and one with cardiac arrest. Hemodynamic and biochemical data before and 1 week after LVAD placement are expressed as mean and range values. None of the patients required right ventricular assist, and all patients achieved functional recovery while on LVAD support (8-71 days). Currently, all four patients are alive (11-22 months) after successful CTx. We conclude that emergency implantation of an LVAD in adolescents with biventricular heart failure can be life saving. As has been shown in the adult population, such a ventricular assist system restores normal circulatory hemodynamics, reverses multi-organ dysfunction, and provides a "safe" bridge to transplantation. PMID- 11016510 TI - In vitro model for studying the effects of hemodynamics on device induced thromboembolism in human blood. AB - Biomateria related thromboembolism is a complex phenomenon, affected by such variables as biomaterial surface chemistry, hemodynamics, and individual donor variations. Thus, isolation of the individual variables would greatly facilitate the understanding and inhibition of this phenomenon. A low volume in vitro model with this potential has been developed, with the initial focus on studying the influence of hemodynamics on thromboembolism (TE) in human blood. Patterned after a larger in vitro model for bovine blood used successfully in our laboratory, the smaller model directed fresh human blood in a single pass through 1/32 inch ID PVC tubing and a flow cell at 3 ml/min. The flow cell consisted of alternating abrupt expansions and contractions of cylindrical tubing that could be modified to study the effects of hemodynamic parameters on TE. Thrombus growth in the flow cell was monitored visually by transillumination microscopy. Emboli from the flow cell were detected continuously by a light-scattering microemboli detector (LSMD), and their strength was assessed by using the constant-pressure filtration (CPF) method. Preliminary studies confirmed the potential of this model. Thrombi were observed visually in the flow cell at sites of high vorticity and at flow separation and reattachment points and were also observed to embolize. Emboli were detected by the LSMD downstream of the flow cell in significantly greater numbers than upstream and were coincident with the embolization of thrombi observed visually. Emboli collected downstream of the flow cell occluded the CPF filters at 50 mm Hg, suggesting that they possessed sufficient strength to occlude microvessels. This model may be used to aid in developing a computer model of thromboembolism, which could subsequently be refined with clinical data. PMID- 11016511 TI - Initial experience with the AbioCor implantable replacement heart at the University of Louisville. AB - Potential benefits of heart transplantation are limited by the severe donor organ shortage. The AbioCor implantable replacement heart has been developed as a potential alternative to heart transplantation. We report our initial experience with the AbioCor in a bovine model. A right thoracotomy was performed for access to the heart and great vessels. After initiation of cardiopulmonary bypass, excision of the native ventricles was followed by orthotopic placement of the IRH and complete implantation of the transcutaneous energy transfer coil, controller, and battery pack. Invasive monitoring of IVC, SVC, carotid artery, pulmonary artery, and left atrial (LA) pressures was performed in all animals. Twelve calves have undergone implantation of the AbioCor. There were three early deaths, one from bleeding, one from respiratory failure, and one from neurodysfunction from low flow during CPB. Nine animals have had a normal recovery and survived a mean of 24.5 days (range, 4-48 days). All the animals have demonstrated excellent hemodynamics with the maintenance of normal pressures in the LA, SVC, IVC, pulmonary artery, and aorta. Adjustment of the right-sided internal hydraulic fluid shunt has allowed for control of right-left balance and, thereby, manipulation of left and right side filling pressures. Late morbidity has consisted of neck wound infection and sepsis, pneumonia, and bleeding. Successful orthotopic implantation of all components of the AbioCor has been achieved in a bovine model. This device has demonstrated restoration of normal hemodynamics and excellent function of the atrial hydraulic shunt to achieve right-left balance. PMID- 11016513 TI - Cost-benefit analysis of extended antifungal prophylaxis in ventricular assist devices. AB - This report defines the cost and benefit of extended antifungal prophylaxis in ventricular assist device (VAD) patients (pts). Extended antifungal prophylaxis is defined as prophylaxis with fluconazole or nystatin that is given until pts are extubated and off antibiotics. These data are compared with that obtained from earlier VAD patients who only received anti-fungal drugs for documented fungal colonization or infection. Thirty-six patients had HeartMate (n = 15) or Thoratec (n = 21) VADs between 1989 and 1997. Cultures positive for fungus (n = 52 cultures) were obtained from 16 of 36 patients (44% of patients). Forty-three fungal cultures were in the preprophylaxis and nine in the postprophylaxis era. There was one death attributable to fungal sepsis in the preprophylaxis era and none in the postprophylaxis era. The total cost of antifungal drugs in the preprophylaxis era was $3,840 over 1,498 patient days (PD) (mean $2.56 per PD), versus $70,670 over 1,525 PD in the postprophylaxis era (mean $46.34 per PD). Extended antifungal prophylaxis was not cost effective in VAD patients at this institution. However, short-term perioperative antifungal prophylaxis was not addressed by this study. We are now using short-term antifungal prophylaxis with fluconazole and nystatin in VAD patients because of the potential for serious morbidity and mortality that is associated with fungal device infections. A future analysis will determine the usefulness of this change in strategy. PMID- 11016512 TI - Influence of pulsatile and nonpulsatile left heart bypass on the hormonal circadian rhythm. AB - We investigated endocrine circadian rhythm (CR) during pulsatile and nonpulsatile left heart bypass (PLHB and NLHB). A ventricular assist device (VAD) was installed in five goats. After 2 weeks of PLHB, NLHB was subsequently conducted for 4 weeks. The levels of melatonin, cortisol, and renin activity were measured every 2 hours for 24 hours on the last day of the second PLHB week and the second and fourth NLHB weeks. Acquired data were compared with those of the normal control goats (Cont) to determine the presence or absence of CR, as well as the timing, acrophase, amplitude, and mesor of CR. Variations in melatonin, CR were consistently detected at all sampling points. In contrast, CR of cortisol during PLHB and NLHB was observed in a few cases, although considerable CR was noted in most of the Cont cases. The variations in renin activity indicated CR in most of the Cont cases and in all cases at all sampling points during PLHB and NLHB, whereas their acrophases were diversified. In conclusion, the CR of melatonin, reflecting the fundamental circadian clock, was maintained during both PLHB and NLHB, while it could be modified for stress sensitive cortisol and hemodynamic responsive renin during these bypass modalities. PMID- 11016514 TI - Noninvasive assessment method to determine the anatomic compatibility of an implantable artificial heart system. AB - To assess the anatomic compatibility of an artificial heart (AH), we attempted to develop a computer environment that would facilitate a reliable simulation of an AH implanted in the human thorax. A three-dimensional thoracic computer model with a ventricle-resected heart was constructed, by using manually extracted contour points of the aorta, pulmonary artery, atria, atrioventricular valves, diaphragm, and thoracic wall from a set of consecutive CT images. Such a model enabled simulation of an AH implantation by orienting the AH model in it. Error evaluation on CT imaging and contour extraction with a Plexiglas cylindrical phantom showed that the diameter of the extracted phantom contour was approximately 2 mm smaller than its original with a standard deviation of <0.5 mm. Errors in contour and surface reconstruction could be reduced to far less than 1 mm under constrained conditions. A study on the influence of breathing revealed that variations in some thoracic dimensions between inspiration and expiration could reach 10 mm. In summary, computer simulation of AH implantation is a worthwhile approach with acceptable accuracy, although further considerations of extreme thoracic situations will be required. PMID- 11016515 TI - Relationship of blood pressure and pump flow in an implantable centrifugal blood pump during hypertension. AB - The purpose of this study was to evaluate the real time relationship between pump flow and pump differential pressure (D-P) during experimentally induced hypertension (HT). Two calves (80 and 68 kg) were implanted with the EVA-HEART centrifugal blood pump (SunMedical Technology Research Corp., Nagano, Japan) under general anesthesia. Blood pressure (BP) in diastole was increased to 100 mm Hg by norepinephrine to simulate HT. Pump flow, D-P, ECG, and BP were measured at pump speeds of 1,800, 2,100, and 2,300 rpm. All data were separated into systole and diastole, and pump flow during HT was compared with normotensive (NT) conditions at respective pump speeds. Diastolic BP was increased to 99.3+/-4.1 mm Hg from 66.5+/-4.4 mm Hg (p<0.01). D-P in systole was under 40 mm Hg (range of change was 10 to 40 mm Hg) even during HT. During NT, the average systolic pump flow volume was 60% of the total pump flow. However, during HT, the average systolic pump flow was 100% of total pump flow volume, although the pump flow volume in systole during HT decreased (33.1+/-5.7 vs. 25.9+/-4.0 ml/systole, p<0.01). In diastole, the average flow volume through the pump was 19.6+/-6.9 ml/diastole during NT and -2.2+/-11.1 ml/diastole during HT (p<0.01). The change in pump flow volume due to HT, in diastole, was greater than the change in pump flow in systole at each pump speed (p<0.001). This study suggests that the decrease of mean pump flow during HT is mainly due to the decrease of the diastolic pump flow and, to a much lesser degree, systolic pump flow. PMID- 11016516 TI - Hemodynamics on abrupt stoppage of centrifugal pumps during left ventricular assist. AB - A magnetically suspended centrifugal pump (MSCP), developed for long-term ventricular assist, is reliable and durable because it has no shaft or seal. However, with nonvalve pumps such as a MSCP, regurgitation occurs when they accidentally stop without cannula clamping. We investigated the hemodynamics during temporary stoppage of a MSCP being used as a left ventricular assist system (LVAS), comparing two inflow cannulation sites. In four sheep (weight, 35 45 kg), microspheres were injected into the left main coronary artery to induce heart failure. An outflow cannula was sutured onto the descending aorta, and two inflow cannulae were inserted into the left atrium and the left ventricle. The MSCP was stopped with both the left ventricular cannula and left atrial cannula clamped, and the hemodynamics and P-V loops were recorded. Each cannula was then unclamped in order, and similar parameters were recorded. LVEDP increased at unclamping of the left ventricular cannula (ULVC), and rose further at unclamping of the left atrial cannula (ULAC). Aortic pressure did not change at ULVC, but decreased at ULAC. The effective systemic flow that subtracted the regurgitant flow through the MSCP from left ventricular output was half at ULVC and almost 0 at ULAC. When stopping centrifugal pumps without circuit clamping, hemodynamic deterioration is less at ULVC than at ULAC. This finding suggests that left ventricular inflow cannulation is recommended to allow more time in emergency situations. PMID- 11016517 TI - Computer modeling of interactions of an electric motor, circulatory system, and rotary blood pump. AB - The innovative ventricular assist systems (IVAS) is the next generation ventricular assist device for use as a permanent implantable device. Its practical application depends upon control of the electric motor and interactions of the electric motor, blood pump, and circulatory system. Computer modeling and simulation are necessary to investigate and evaluate the interactions and feasibility of sophisticated control algorithms. In this paper, a computer model of the complete system, including the cardiovascular system, blood pump, and electric motor, is proposed. The model is obtained based on an electric circuit model of the cardiovascular system, a parametric model of the blood pump, and a dynamic model of the electric motor. The cardiovascular system uses nonlinear parameters to simulate the time-varying property of the ventricles, and the cannula collapse effect caused by over-pumping. The blood pump model can be obtained either from pump design data, or test data. The motor control can be operated with closed-loop regulation, depending upon physiologic requirements. Different operation modes (current or speed) of the electric motor can be simulated. The computer model is implemented using MATLAB. Various motor operation modes are simulated and their effects are evaluated. By adjusting the motor input, the pump can achieve proper output so that normal physiology can be obtained. In addition to evaluating existing operation modes and their effect on the physiologic system, the computer simulation results show that this computer model can contribute significantly to the development of new physiologic control algorithms. It is demonstrated that, using this motor-pump-physiology interaction model, development of an innovative ventricular assist system can be greatly facilitated. PMID- 11016518 TI - Estimate of gas transfer rates of an intravascular membrane oxygenator. AB - An asymmetric hollow fiber membrane was prepared from a newly synthesized fluorinated aromatic polyimide (6FDA-6FAP) by using a dry/wet phase inversion process. The membrane was used in a membrane oxygenator over a long period of time. In this study, the potential of the membrane for intravascular membrane oxygenation (IVOX) was studied in respect to oxygen transfer. The gas permeance of the membrane and three commercially available hollow fiber membranes for membrane oxygenators was measured in a gas-gas system and a gas-liquid system and discussed relative to the membrane structures. The oxygen transfer rates of the IVOX devices using these four membranes were estimated by a mathematical kinetic model, with the oxygen permeance measured in the gas-liquid system. The results showed that the device using the 6FDA-6FAP hollow fiber membrane has the highest oxygen transfer rate and is believed to be applicable to IVOX. The methods to determine oxygen transfer rate of a hollow fiber membrane and the mathematical kinetic model, are useful for developing a hollow fiber membrane and a device for oxygenation. PMID- 11016519 TI - Insensible water loss during extracorporeal membrane oxygenation: an in vitro study. AB - To measure insensible fluid loss from silicone membrane oxygenators during extracorporeal membrane oxygenation (ECMO), an in vitro system was used. A standard neonatal ECMO circuit (Avecor) was connected to a noncompliant reservoir, which was then primed with normal saline. The experiment was conducted by using two silicone oxygenators (Avecor 0.4 and 0.8 m2), three gas flow rates (0.5, 1.0, and 2.0 L/min) (sweep), and two fluid flow rates (200 and 400 ml/min). Two methods were used to measure the water loss. One method was to replace the water to the noncompliant circuit by using a calibrated burette, and the other method was to collect condensed water after cooling the postmembrane sweep gas to 0 degrees C. The influence of the amount of sweep, fluid flow rate, size of membrane, and inlet and outlet sweep gas temperatures on measured water loss was statistically determined. The amount of water loss correlated with sweep (r2 = 0.81; p<0.00001) but was not related to the fluid flow rate, membrane size, or inlet and outlet sweep gas temperature. The average daily fluid loss measured with replacement and collection methods for each liter of sweep per minute were 72.0+/-12.6 and 62.3+/-10.0 ml, respectively. This information may be applied to clinical practice to accurately manage fluid balance in the sick neonate on ECMO. PMID- 11016520 TI - Role of cellulosic and noncellulosic membranes in hyperhomocysteinemia and oxidative stress. AB - Hyperhomocysteinemia is an independent risk factor for cardiovascular morbidity and mortality in end-stage renal disease (ESRD) with an increased relative risk (RR) of 1% per micromol/L in total homocysteine concentration. In ESRD patients who undergo hemodialysis (HD), the antioxidant system is largely inadequate in correcting the imbalance between generation and scavenging of reactive oxygen species (ROS). To clarify the role of several cellulosic (CMs) and noncellulosic of synthetic membranes (NCMs) upon hyperhomocysteinemia and the oxidative stress, we measured plasma (P) homocysteine (t-HCY), plasma lipid peroxidation (LPO), and erythrocyte (E) concentration of several antioxidant enzymes in 20 normal subjects, in 35 HD patients treated with CMs, and in 29 patients treated with NCMs. Before, during, and after the first session of the week (at times 0', 120', end, 30' after HD end), blood samples were drawn. Plasma (P) homocysteine (t HCY), cysteine (CYS), malondialdehyde (MDA), erythrocyte (E)-glutathione (GSH), glucose-6-phosphodehydrogenase (G6PD), glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), and superoxide-dismutase (SOD) were determined. The dialytic procedure significantly decreased the three plasma parameters, but none normalized (as a mean). The E-enzymes scavenging ROS (lower than normal before session) increased throughout the session, but the normal range of activity was never reached. Different membranes have shown different effects. When these effects on P and E spaces were pooled, we were able to classify the membranes as follows. In a general sense, cellulosic membranes are less effective than synthetic membranes both on lipoperoxides (LPO) and antioxidant activity (AOA). Among synthetic membranes, PMMA is the best membrane both for plasma values and lesser enzymatic derangement during the session. A practical system for classifying the anti-atherosclerotic action and antioxidant activity of dialytic membranes is proposed. PMID- 11016521 TI - Phosphodiesterase III inhibitor olprinone chlorate is not significantly removed by continuous venovenous hemodiafiltration. AB - The study objective was to determine whether a phosphodiesterase III inhibitor, olprinone chlorate, is effectively removed by continuous venovenous hemodiafiltration (CVVHDF) in a patient with cardiac and renal failure. The patient was a 73 year old man who had undergone coronary artery bypass grafting for ischemic heart disease and who developed cardiac and renal failure postoperatively. A 0.2 microg/kg per minute dosage of olprinone chlorate was administered intravenously for 120 minutes while the patient was treated with CVVHDF. Samples from the arterial and venous blood catheters and those from the ultradiafiltrate for 12 hours were collected to calculate pharmacokinetic parameters and clearance of hemodiafiltration. The calculated parameters were as follows: half-life of elimination phase: 4.96 hours; total clearance 3.40 ml/min per kg. The clearance of CVVHDF was 0.33 ml/min per kg. The olprinone chlorate clearance of CVVHDF approximates only 10% of total clearance in this case. CVVHDF may not produce significant reduction in the serum olprinone chlorate level. It is recommended that the infusion dosage of olprinone chlorate should be reduced when given to patients with renal failure even if treated with CVVHDF. PMID- 11016523 TI - Hospitalization for suicide attempt and completed suicide: epidemiological features in a managed care population. AB - BACKGROUND: Understanding factors that contribute to high suicide risk holds important implications for prevention. We aimed to examine the sociodemographic and medical predictors of attempted suicide (severe enough to require hospitalization) and of completed suicide in a large population-based sample from a health maintenance organization (HMO) in northern California, USA. METHOD: We designed a cohort study, including 87,257 women and 70,570 men aged 15 through 89 years old at baseline (in 1977-1985) with follow-up for hospitalizations and mortality through the end of 1993. RESULTS: After a median of 10 years, 169 first hospitalizations for attempted suicide (111 among women, 58 among men) and 319 completed suicides (101 among women, 218 among men) were identified. There was a greater incidence of hospitalization for suicide attempt in women than in men and, conversely, a greater incidence of completed suicide in men than in women. The predominant methods of attempted and completed suicides were ingestion of psychotropic agents and use of firearms, respectively. In gender-specific multivariate analysis of hospitalization for suicide attempt, statistically significant associations were seen for age 15-24 years (women), 65-89 years (men), white race (women), 12th grade or less education (both genders), technical/business school education (men), never being married (men), history of emotional problems (both genders), history of family problems (women), history of job problems (men) and presence of one or more comorbidities (men). The independent predictors of completed suicide were: age 15-24 years (both genders), Asian race (women), Caucasian race (both genders), never being married (both genders), being separated/divorced (women), prior inpatient hospitalization for suicide attempt (both genders) and history of emotional problems (both genders). CONCLUSION: These findings could help health professionals be more effective in the prevention of suicide morbidity and mortality. PMID- 11016522 TI - Early detection and assertive community treatment of young psychotics: the Opus Study Rationale and design of the trial. AB - BACKGROUND: Recent research indicates that early detection of young persons suffering from psychosis and subsequent intensive intervention enhances treatment response and prognosis, but the data are only preliminary and suggestive. METHOD: We present the rationale and design of the largest study to date to evaluate two major issues in the field of secondary prevention: (1) Does education and intensified collaboration with general practice, social services etc. reduce the duration of untreated psychosis? and (2) Can modified assertive community treatment improve the course and outcome in young persons suffering from psychosis as compared to treatment in community mental health centres? The article aims additionally to put the study in context and assist in designing future studies. RESULTS: Preliminary experiences are described. The findings of the first 312 patients show that modified assertive community treatment results in patients adhering to treatment significantly better than standard treatment in community mental health centres. CONCLUSION: The surge of interest in preventively oriented detection and treatment models for untreated psychosis in young people calls for research programmes and evidence. The obstacles to this are manifold. The initial findings of the OPUS study suggest, however, that better adherence to treatment is possible. PMID- 11016524 TI - Dual diagnosis in the suburbs: prevalence, need, and in-patient service use. AB - BACKGROUND: Previous research has found comorbid severe mental illness and substance misuse (dual diagnosis) to be highly prevalent and to be associated with serious clinical and social problems, and increased service use in inner city populations. The present study measures the prevalence of dual diagnosis, patterns of substance misuse, and associated in-patient use in a more demographically representative population in a suburban area of South London. METHOD: We identified representative prevalent cases with psychotic illnesses who had been in contact with services in a geographically defined catchment area in Croydon over the previous 6 months. Cases of alcohol or substance misuse and dependence were identified through standardised interviews with patients and keyworkers, and sociodemographic and in-patient psychiatric service use data were also recorded. RESULTS: Sixty-one of the 124 cases identified were randomly selected for interview, of whom 66% responded (N = 40). The prevalence rates of dual diagnosis (DD) observed were 33% (95% CI 18-47%) for any substance misuse, 20% (95% CI 8-32%) for alcohol misuse only, 5% (95% CI-16 to 26%) for drug misuse only, and 8% (95% CI -0.7 to 16%) for both drug and alcohol misuse. A lifetime history of any illicit drug use was observed in 35% of the sample (95% CI 20 50%). Patients who misuse alcohol and drugs were not found to be more likely to have been admitted to hospital in the previous 2 years, with little difference being observed between DD and psychosis-only patients in the mean number of in patient admissions in this period (mean difference 0.25, 95% CI for difference 1.5 to 2.0). However, the DD patients were found to have spent on average over twice as long in hospital as other psychotic in-patients over the previous 2 years (mean difference 67.3 days, 95% CI for difference -205.9 to 71.2 days). DD patients were also found to have a greater number of unmet areas of need than the psychosis-only patients, which included accommodation, daytime activity, and social life, as well as substance misuse. CONCLUSIONS: The prevalence of substance misuse in patients with severe mental disorders in a suburban area is about as high as that for similar patients in inner-city London. While DD patients are not admitted more often than patients with psychosis alone, they have double the length of in-patient stay, which may be attributable to higher levels of unmet need. PMID- 11016525 TI - Diagnostic value of the DSM and ICD categories of psychosis: an evidence-based approach. UK700 Group. AB - BACKGROUND: It is generally assumed that the distinction between affective and non-affective psychosis occasioned by modern diagnostic criteria provides a useful symptomatic contrast. METHOD: In a sample of 708 patients with chronic psychosis, the distinction of lifetime DSM-III-R and ICD-10 diagnoses of affective versus non-affective psychosis was used as a diagnostic test to detect lifetime presence of depressive, manic, positive, negative and disorganisation symptoms. RESULTS: A manic or depressive affective diagnosis was a perfect test to diagnose the presence of manic and depressive symptoms, as evidenced by very high diagnostic likelihood ratios. However, this test result was based solely on the inclusion criterion that patients with affective psychosis must have affective symptoms (guaranteeing high specificity and high likelihood ratios), and ignored the fact that patients with non-affective psychosis also had high affective symptom scores (low sensitivity). Furthermore, a non-affective psychotic diagnosis was a very poor test to diagnose correctly the presence of positive, negative and disorganisation symptoms in comparison with an affective psychotic diagnosis. In general, the DSM-III-R categories performed somewhat better as a diagnostic test than those of ICD-10. CONCLUSION: The evidence for true diagnostic value of the distinction between affective and non-affective psychotic diagnoses is weak. Rather, the distinction appears to obscure natural overlap between the symptom dimensions of the different diagnostic categories. PMID- 11016526 TI - Matching resources to care: the acceptability, validity and inter-rater reliability of a new instrument to assess severe mental illness (MARC-1). AB - BACKGROUND: Most definitions of severe mental illness (SMI) are categorical and assign the patient to either SMI or not-SMI status. While this is useful for some purposes, it is a rather limited approach. The purpose of the present study is to develop a new method of addressing the issue of 'severity', and to develop a dimensional rather than a categorical approach. The paper reports on the acceptability, reliability and validity of a method developed to collect a standard set of data covering the majority of items specified in the academic and policy literature as characterising SMI. METHOD: A single page form, Matching Resources to Care (MARC-1), containing most of the items used in definitions of SMI was used to collect data from community mental health staff about their current open caseload, in four co-terminous health and social services settings during a census week (n = 2139). In addition to the data from the four pilot sites, we conducted a substudy (n = 91), in which two raters rated the same cases during the same week. RESULTS: The MARC-1 scores were able to distinguish between patients in receipt, and those not in receipt, of specific types of community care (level of care, eligibility for care and statutory aftercare) (P < 0.001). The MARC-1 score was modestly but significantly correlated (r = 0.28) with the Global Assessment Scale (P < 0.001). The mean percentage inter-rater agreement for the MARC-1 score items was 87%. CONCLUSION: It is possible to use a simple census form in both health and social services agencies. The completion rates were good in both services. The levels of reliability were good, and concurrent validity was established with specific types of care in the community. PMID- 11016527 TI - Psychometric properties of the Parental Bonding Instrument and its association with psychological distress in a group of incarcerated young offenders in Scotland. AB - BACKGROUND: The Parental Bonding Instrument (PBI) was developed in order to help quantify the parental contribution to psychological distress. It has subsequently been shown to be of value in examining the influence of parents in many diverse areas including delinquency. METHOD: A shortened form of the PBI and the Hospital Anxiety and Depression Scale (HADS) were administered to a group of young offenders held in custody in Scotland. RESULTS: High levels of psychological distress were linked with low parental care, but there were no associations between psychological distress and parental control. However, an analysis of the factor structure found that, although the care factor of the PBI showed good internal reliability, the control factor was less well defined in this group. CONCLUSIONS: It is suggested that a three-factor solution representing care, control with regard to independence and protective control may be more appropriate. PMID- 11016528 TI - A cross-cultural study of mental health beliefs and attitudes towards seeking professional help. AB - BACKGROUND: This study sets out to examine the relationship between culture beliefs about the causes of mental distress and attitudes associated with seeking professional help for psychological problems. It was hypothesised that there is a meaningful and statistical relationship between these variables and that there will be a difference in this relationship between Asians and Westerners. Participants were 287 adults belonging to three groups (British Asian, western European and Pakistanis). METHOD: Participants completed two questionnaires: the Orientations to Seeking Professional Help (Fischer and Turner 1970) and the Mental Distress Explanatory Model Questionnaire (Eisenbruch 1990) and a demographic data sheet. RESULTS: Analysis indicated that positive attitudes toward seeking professional help for psychological distress were similar for British Asians, Westerners and Pakistanis. There were significant differences between the three groups in the causal attributions of mental distress. Although culture, as a variable, was not a significant predictor of a positive attitude to seeking professional help, causal beliefs of mental distress were significant predictors of attitudes to seeking help for the British Asian and the Pakistani groups. Beliefs were not significant predictors for attitudes to seeking help for the Western group. CONCLUSION: It was concluded that culturally determined causal beliefs of mental distress contribute to attitudes towards seeking professional help for psychological problems for Asians. Implications for both research and the provision of more appropriate health services for the British Asian minority group in the United Kingdom are discussed. PMID- 11016529 TI - LTP induction threshold change in old rats at the perforant path--granule cell synapse. AB - Old, memory-deficient rats do not show a change in the threshold for long-term potentiation (LTP) induction in hippocampal region CA1. This observation suggests that defective NMDA receptor mechanisms at the Schaffer collateral-CA 1 pyramidal cell synapse cannot explain age-related LTP induction deficits that are observed under some stimulation protocols. The effects of aging on functional electrophysiology are not, however, identical between hippocampal subregions. In fact, at the perforant path-granule cell synapse of rats NMDA receptor-mediated responses are reduced, suggesting a possible change in the threshold for LTP induction at this synapse. This hypothesis was tested in the present experiment. We found that when weak orthodromic stimulation of medial perforant path fibers is paired with intracellular current injection of granule cells, the threshold for LTP induction is elevated in aged, spatial memory-impaired rats compared to middle-aged and young controls. Thus, in addition to there being fewer total medial perforant path synaptic contacts in old rats, greater depolarization and input convergence is required before durable modification of synaptic strength can be induced. PMID- 11016530 TI - The effects of aging on the frequency of nerve fibers in rhesus monkey striate cortex. AB - In the rhesus monkey primary visual cortex, there are bundles of vertically oriented myelinated axons, which mainly contain efferent fibers originating from pyramidal cells. At the level of layer 4Cbeta, the bundles are regularly arranged and the nerve fibers in them are closely packed. In order to determine if a significant loss of intracortical nerve fibers occurs as the primate cerebral cortex ages, the frequency of vertically oriented myelinated fibers was examined at the level of layer 4Cbeta in 1 microm-thick, tangential sections. The results show no statistically significant differences in the numbers of vertically oriented fibers beneath 1 mm(2) of cortical surface between young, middle-aged, and old monkeys, and electron microscopic examination reveals few signs of degenerating axons. There is, however, an age-related breakdown of the myelin in sheaths that surround some axons. Thus, the data indicate that there is not a loss of vertically oriented myelinated fibers from the cortical gray matter during aging, although their sheaths may be altered. PMID- 11016531 TI - Sympathetic neurite growth on central nervous system sections is region-specific and unaltered by aging. AB - Several lines of evidence suggest that the brain exhibits reduced plasticity with aging. However, a variety of soluble neurite outgrowth-promoting factors, such as neurotrophins, are not decreased in the aged brain, and aged neurons do not possess dramatically reduced growth potential. The possibility that aging results in reduced baseline substrate-bound neurite outgrowth-promoting activity in the central nervous system (CNS) was evaluated using tissue section culture. There were clear differences between brain regions in the extent of neurite outgrowth on both young and aged brain sections. However, no differences in the extent of neurite outgrowth were observed as a function of age. These results suggest that aging of the rat CNS is not accompanied by major alterations in the baseline neurite outgrowth-promoting substrate properties of the tissue. PMID- 11016532 TI - Calorie restriction increases light-dependent photoreceptor cell loss in the neural retina of fischer 344 rats. AB - We investigated the effect of > or = 8 months of 40% caloric restriction (CR) on photoreceptor cell loss in 12, 18, and 24 month-old Fischer 344 rats (N = 154). Rats were reared at the NIA Biomarkers Program, National Center for Toxicological Research. Photoreceptor cell density, assessed histologically, declined with age in both the CR-fed and ad lib (AL)-fed cohorts (P < 0.000), but declines were more pronounced in the CR cohort (P < 0.0005). The deleterious effect of CR was most pronounced in the central as opposed to the peripheral retina (P = 0.008), suggesting a light-dependent mechanism. Photoreceptor cell density was inversely associated with rearing under bright light (300-750 lux) as compared with rearing under lower illuminance (< or = 200 lux) (P < 0.0005). However, the deleterious effect of bright light on photoreceptor cell density was more pronounced in the CR cohort (P = 0.04). Effects of CR on circadian activity are likely to increase the actual light exposure of the CR cohort and may explain the apparent inability of CR to delay retinal aging in albino rats. PMID- 11016533 TI - Inflammatory 5-LOX mRNA and protein are increased in brain of aging rats. AB - 5-Lipoxygenase (5-LOX) is the key enzyme in the synthesis of leukotrienes, inflammatory mediators of arachidonic acid. 5-LOX is also expressed in neurons (in particular in the hippocampus and the cerebellum), and it seems to be capable of promoting neurodegeneration. Recently, we observed greater 5-LOX mRNA content in the hippocampus of older (24 months) than younger (2 months) rats. In this study, we measured in the hippocampus and the cerebellum of younger and older male F344 rats the contents of: 5-LOX mRNA, FLAP (5-LOX activating protein) mRNA, and 5-LOX protein. By using a quantitative reverse transcription/polymerase chain reaction (PCR) (RT-PCR) with internal standards we found that 5-LOX but not FLAP mRNA content is greater (both in hippocampus and cerebellum) of older than younger rats. By using quantitative Western immunoblotting, we found a greater content of 5-LOX protein in the hippocampus and the cerebellum of older rats; we also established that the membrane/cytosol 5-LOX content ratio is larger in the brains of older than younger rats (statistically significant in the cerebellum). The latter can be considered an indication of 5-LOX translocation/activation during aging. Together these results suggest that aging increases both neuronal 5 LOX expression and protein translocation, and indicate that the 5-LOX system might play a significant role in the pathobiology of aging-associated neurodegenerative diseases. PMID- 11016534 TI - Age-related changes in ethanolamine glycerophospholipid fatty acid levels in rat frontal cortex and hippocampus. AB - Morphological and biochemical alterations are associated with a progressive age related cognitive deficit. Plasmenylethanolamine, the major brain plasmalogen, may be modified during aging because of a possible antioxidant role and involvement in synaptic transmission. Two- and 18-month-old rats were used to study the effect of aging on the levels and acyl composition of plasmenylethanolamine (PmE), phosphatidylethanolamine (PE), and phosphatidylserine (PS) in the frontal cortex and hippocampus. Aging only reduced significantly the PE levels in the frontal cortex. In 18-month-old rats, the fatty acid composition of the three phospholipid classes studied showed an increase of monounsaturated fatty acid (18:1 n-9 and 20:1 n-9) and a decrease in polyunsaturated fatty acid (PUFAs), essentially docosahexaenoic acid (DHA). DHA was markedly decreased in hippocampus PE. DHA, but also arachidonic acid, were considerably lower in frontal cortex PmE. PS modifications were similar in both regions. Hippocampus and frontal cortex underwent specific age-induced modifications in PmE and PE acyl composition. This could produce different effects on the functional ability of these two structures involved in the processes of specific memorization. PMID- 11016535 TI - Age-related changes of apoptotic cell death in human lymphocytes. AB - Apoptosis seems to be involved in immunosenescence associated with aging. Moreover, in lymphocytes (PBL) of patients with Alzheimer's disease, an increased susceptibility to the apoptotic pathway has been described possibly due to impaired protection of oxidative stress. Accordingly, it seemed to be of particular interest to investigate the contribution of normal aging to the susceptibility from human lymphocytes to programmed cell death. We could show that PBL from elderly individuals (>60 years) accumulate apoptosing cells to a significant higher extent in spontaneous and activation-induced cell death compared to younger controls (<35 years). Treatment with the oxidative stressor 2 deoxy-D-ribose or with agonistic-CD95-antibody pronounced this effect even more implicating a higher sensitivity to reactive oxygen species and a higher functional CD95 expression, respectively. In addition, expression of the activation markers HLA-DR and CD95 was significantly increased in CD3+-cells of aged subjects, while expression of CD25 did not seem to be affected by age. Expression of Bcl-2 was increased in aging and correlated with the number of apoptotic cells. PMID- 11016536 TI - Age-related decline in water maze learning and memory in rats: strain differences. AB - Rats display an age-related impairment in learning and memory; however, few studies have systematically examined this relationship in multiple strains. The present study used a repeated acquisition water maze task to test the hypothesis that age-related decreases in learning and memory occur at different rates in three strains of rats, i.e. Sprague-Dawley (SD), spontaneously hypertensive (SHR), and Wistar Kyoto (WKY) rats. All three strains of rats displayed age related decreases in spatial learning and memory; however, the rate of decline differed between the strains. Compared to young rats of the same strain, only SHR were significantly impaired at 12 months of age. All three strains displayed moderate impairment in learning the task at 18 months of age, and at 24 months of age all three strains of rats were severely impaired in the task, but SD performed best at 18 and 24 months of age. Further, SD and SHR displayed a probe trial bias at 3 months of age, but only SD had a bias at 12 months of age and none of the rats showed the bias at later ages. Thus, in these three strains, age related impairment of spatial memory proceeds at different rates. PMID- 11016537 TI - Age-related dopamine D2/D3 receptor loss in extrastriatal regions of the human brain. AB - Loss of dopamine D2-like receptors in the striatum has been associated with both normal human aging and impairment of cognitive and motor functions in the elderly. To investigate whether there are age-associated changes in dopamine D2 and D3 receptor subtypes (D2/3Rs) outside the striatum, a D2/3R selective high affinity radioligand [11C]FLB 457 was used in positron emission tomography (PET) examinations for 24 normal healthy male subjects (age range 19-74 years). Significant age-related declines of D2/3Rs were detected in all the brain regions studied: the anterior cingulate cortex (decline of 13% per increase of a decade in age, P < 0.001). the frontal cortex (11%, P < 0.001), the lateral temporal cortex (10%, P < 0.001), the hippocampus (10%, P < 0.01), the medial temporal cortex (9%, P < 0.001), the amygdala (7%, P < 0.01), the medial thalamus (6%, P < 0.001) and the lateral thalamus (5%, P < 0.01). The rate of D2/3R decline was significantly faster in the frontal cortex as compared to the medial temporal cortex (P < 0.05, Bonferroni corrected) and as compared to the medial thalamus (P < 0.05, Bonferroni corrected). These results indicate that the previously demonstrated age-related decline in striatal dopamine D2 receptors extends to several extrastriatal regions in normal human males. Further, the rate of D2/3R decline may be faster in the frontal cortex as compared to the temporal and thalamic regions. PMID- 11016538 TI - Age-dependent changes in recovery sleep after 48 hours of sleep deprivation in rats. AB - To characterize possible changes in homeostatic regulation of sleep with aging, we have examined sleep stages during recovery sleep after 48 h of sleep deprivation in young (3 months), middle aged (12 months), and old (24 months) rats. It was found that young and middle aged, in contrast to old rats, had large (21-24%) increases in total sleep time during recovery sleep; the old rats experienced a quantitatively small (8%) but significant rise in total sleep. NREM sleep increased significantly during the recovery period in young and middle aged, but not older rats. High voltage NREM sleep (HS2) declined by 30% during recovery in the young animals, but remained unchanged compared to baseline in the middle aged and old animals. The young and middle aged rats had increases in REM sleep during recovery compared to their baseline by 96% and 93%, respectively, which was significantly greater than a 65% increase during recovery in the old rats. Increases in total sleep and REM sleep during recovery were largely confined to the first 6 h in young and middle aged rats, but maxima for the old rats occurred in the second 6 h. PMID- 11016539 TI - Vascular and parenchymal Abeta deposition in the aging dog: correlation with behavior. AB - The behavior of 25 dogs was indirectly assessed by a formal questionnaire (evaluation of Age-Related Cognitive and Affective Disorders-ARCAD), filled out by the owner. The density of diffuse and vascular deposits was evaluated using four anti-Abeta peptide antibodies, in four temporal areas. Parenchymal, diffuse deposits of Abeta42 peptide were found in all aged animals but one. They were Congo red negative and were not immunostained by the anti-Abeta40 antibody, contrary to the vascular deposits. The densities of vascular and parenchymal deposits were not correlated. The ARCAD score was correlated with age, density of Abeta parenchymal and vascular deposits, and with the number of areas containing deposits (extension index). Multivariate analysis showed that the age and the extension index explained most of the variance. Congo red positivity (indicating that the Abeta peptide has the characteristics of an amyloid substance) is limited in the dog to the vascular wall and is associated, as in man, with the deposition of the Abeta 1-40 isoform. Parenchymal Abeta deposition seems to be a common correlate of behavioral problems in aging dogs. PMID- 11016540 TI - Behavior and brain uptake of fluorodeoxyglucose in mature and aged C57BL/6 mice. AB - This paper describes regional brain energy metabolism, sensorimotor, and memory functions in mature (12 months) and old (24 months) C57BL/6 mice. Male mice were tested across a variety of sensorimotor procedures and in the Morris water maze before evaluating brain uptake of fluorodeoxyglucose (FDG) in the resting state. Mature mice outperformed older mice during the difficult sensorimotor and memory tasks, but not the easier tasks. This suggested that the greater sensorimotor and memory demands of a task compromised the performance of the older mice. This conclusion was consistent with the relative FDG uptake decreases found in regions mediating sensorimotor coordination (vestibular, cerebellar, ventral thalamic regions) and some limbic regions linked to memory (mammillary body, posterior cingulate, and piriform cortices). The inferior colliculus and flocculus had the greatest metabolism in mature mice, as shown by resting FDG uptake, and these regions showed the most marked hypometabolism in the older mice. The data also showed that the neurobehavioral correlative pattern in older mice was modified. In conclusion, brain regions with higher energy metabolism, and the behaviors to which they are related, were affected most greatly by aging in C57BL/6 mice suggesting that these areas are more metabolically vulnerable to aging effects. PMID- 11016541 TI - Conformational change as one of the earliest alterations of tau in Alzheimer's disease. AB - Paired helical filaments (PHFs) found in Alzheimer's disease (AD) are mainly comprised of an abnormal form of tau (PHF-tau) that has undergone several post translational modifications. Previous studies have shown that the monoclonal antibody MCI identifies a distinct conformation of tau in AD. We have assessed the temporal and spatial occurrence of the tau conformation recognized by MC1, and found its appearance in hippocampal neurons vulnerable to neurofibrillary tangle (NFT) formation in Braak Stage I and II cases. Electron microscopy has clearly demonstrated that this conformation precedes the formation of PHF. MC1 immunoaffinity chromatography also has identified a nonfilamentous, soluble pool of this abnormal tau. ELISA and immunoblotting have shown that this material is indistinguishable from that found in NFTs. This soluble component has the ability to self-assemble into PHFs in a concentration-dependent manner. Because the conformational change recognized by MCI appears before the assembly of and is found in PHF, but is not present in the normal brain, we suggest that the formation of the MCI epitope is one of the earliest pathological alterations of tau in AD. PMID- 11016542 TI - Cytosolic calcium alterations in platelets of patients with early stages of Alzheimer's disease. AB - Alterations in calcium homeostasis might be implicated in the neuropathology of Alzheimer's disease (AD). To date it is not clear whether changes in cytosolic calcium level ([Ca2+ ]i) are the result or the cause of pathogenic effects. In platelets of patients with early stages of AD, the basal values of [Ca2+]i in the absence of extracellular Ca2+ were significantly lower in comparison with age matched and young controls. After the addition of 1 mM calcium into the incubation medium the [Ca2+]i markedly increased in platelets of AD patients whereas the increase only to a smaller extent was observed in control age-matched and young subjects. The present study proposes that calcium dysregulation during the whole disease period could not be uniform and according to our results the [Ca2+]i is reduced in the first stages of AD. We suggest that the disturbed calcium homeostasis in AD is an "early defect." PMID- 11016543 TI - Relationship between apoE genotype and CSF beta-amyloid (1-42) and tau in patients with probable and definite Alzheimer's disease. AB - We investigated the usefulness of cerebrospinal fluid (CSF) beta-amyloid42 (Abeta42), beta-amyloid40 (Abeta40) and tau analyses in the diagnosis of Alzheimer's disease (AD). The study included 41 definite AD cases, 80 patients with probable AD. 27 with other dementias and 39 neurological controls. Abeta42, Abeta340 and tau protein concentrations in CSF were measured of using ELISA assays. Abeta42 levels were decreased and tau increased in AD. Combination of Abeta42 and tau resulted a sensitivity of 50.4% for AD and specificities of 94.8% for controls and 85.2% for other dementias. Ninety-one percent of the patients with Abeta42 below the cutoff value (340 pg/ml) and tau above the cutoff value (380 pg/ml) had AD. AD patients carrying apoE epsilon4 allele had lower Abeta42 (P < 0.005) and higher tau (P < 0.05) levels than those without an E4 allele, and 18 (81.8%) of the 22 AD patients who had normal Abeta42 and tau levels were apoE e4 allele non-carriers. Low Abeta42 and high tau concentration in CSF strongly support the diagnosis of AD. Measurement of Abeta42 may help the early diagnosis of cases at risk for AD such as apoE E4 allele carriers. PMID- 11016544 TI - Nicotinic receptor losses in dementia with Lewy bodies: comparisons with Alzheimer's disease. AB - We sought to delineate differences between alpha7 nicotinic acetylcholine receptor (nAChR) levels in Alzheimer's disease (AD), dementia with Lewy bodies (DLB) and age matched controls, as well as the correlations between alpha7 or non alpha7 nAChR levels and synaptophysin (Syn) or choline acetyltransferase (ChAT) in DLB. Mean bungarotoxin (Bgt) binding was 2.7 - 1.1 for controls, 2.4 +/- 1.0 for AD and 1.4 +/- 0.5 for DLB. There were significant decreases in Bgt binding for the DLB group compared to either controls or AD. Mean epibatidine (Epi) binding was 14.8 +/- 3.2 for controls, 6.3 +/- 3.2 for AD and 7.1 +/- 2.4 fmoles/mg protein for DLB. Epi binding in both the AD and DLB groups was significantly lower than in the controls. Although Syn loss correlated with the decrease in Epi binding in both diseases, declining ChAT levels correlated with Epi binding only in DLB. These data demonstrate a different pattern of nAChR loss in AD and DLB that may, in part, explain some of the differences in the two phenotypes. PMID- 11016545 TI - Regulation of amyloid precursor protein processing by Abeta in human glioma cells. AB - Amyloid precursor protein (APP) is cleaved to neurotoxic/proinflammatory amyloid beta protein (Abeta) or to the neuroprotective secreted alpha-APPs. A balance in APP metabolism may influence the outcome between toxicity and protection to central nervous system (CNS) neurons in Alzheimer's disease. Treatment of U-373 MG astrocytoma cells with aggregated Abeta (1-40) decreases APP secretion into the medium to 10-30% of control values. This decreased secretion appears to be specific for APP since Abeta treatment causes an approximately 2-fold increase in interleukin-8 (IL-8) secretion. Abeta treatment also causes a 4- to 9-fold increase in total cell-associated APP. This increase is due to cellular retention of alpha secretase-cleaved APP and a 2-fold increase in mature full-length APP. These data suggest that deposition of aggregated Abeta may contribute to Alzheimer's-associated neurotoxicity by altering the metabolism of the APP protein. Abeta may exert harmful effects by decreasing the secretion of neuroprotective or neurotrophic APP and, in addition, by increasing intracellular full-length APP; thereby providing increased substrate for generation of amyloidogenic peptide within astrocytes. PMID- 11016546 TI - The French connection: some contributions of French-language research in the post Piagetian era. AB - This article presents French-speaking researchers' contribution to the field of differential developmental psychology. Following a brief review of key Piagetian ideas pertaining to his conceptualization of individual differences, the core of the article traces methodological and theoretical transformations that were necessary for understanding individual differences within a general theory of cognitive development. On a methodological level, French-speaking researchers went from standardizing Piaget's clinical method to constructing developmental scales and operational tests. On a theoretical level, Reuchlin's writings guided Longeot, and several other French (Lautrey and Bideaud) and Genevan (de Ribaupierre and Rieben) researchers into a scientific quest for a genuine integration of differential and developmental psychology. We present an overview of the pluralistic and multidimensional model of cognitive functioning and development that emerged from the work of the French-Swiss team of researchers. Concluding remarks focus on the actual research agendas of researchers interested in resolving the challenging issue of understanding relationships between inter- and intraindividual differences and general tendencies in cognitive development. PMID- 11016547 TI - Full house or Pandora's box? The treatment of variability in post-Piagetian research. AB - This commentary focuses on the conceptual implications of analyses of individual differences in francophone post-Piagetian research. These analyses are viewed as preoccupied by the "American question" of measurement and method, instead of attempting a theoretical account of the issues raised by intraindividual and interindividual variability in development. PMID- 11016548 TI - Is the French connection neo-Piagetian? Not nearly enough! AB - Some general-causal assumptions of current neo-Piagetian research are discussed and compared with those of French European developmentalists. Developmental theory problems for the millennium are highlighted. PMID- 11016549 TI - The social-emotional and cultural contexts of cognitive development: neo Piagetian perspectives. AB - The neo-Piagetian research on individual differences in cognitive development reviewed by Larivee, Normandeau, and Parent suggests that Piaget's theory can be used to explain variability in development. My commentary explores this question further through a discussion of two additional sources of variation in children's cognitive development: social-emotional context and cultural meanings. PMID- 11016550 TI - From individual differences to dynamic pathways of development. AB - A fruitful way to build upon French-language research on development of analogical and propositional processes in logical reasoning tasks is to use dynamic systems tools to describe and analyze relevant developmental pathways. Issues to address include (1) the characteristics of developmental transitions, such as hysteresis; (2) the nature of growth processes, such as hierarchical development or predator-prey interactions; and (3) the construction of effective scales for measuring change in logical reasoning. PMID- 11016551 TI - Domain specificity and variability in cognitive development. AB - There are core-specific and noncore-specific domains of knowledge, but only the core-specific domains benefit from innate skeletal structures. Core skeletal domains are universally shared, even though their particular foci may vary; individuals vary extensively in terms of the noncore domains they acquire. PMID- 11016552 TI - What is the target? AB - Reconciling the differential perspective with the Piagetian perspective is a very difficult task. The Piagetian perspective admits the existence of interindividual differences but interprets them as noise masking the universal logical succession of structures, whereas the differential perspective views development as consisting of "vicarious processes." As a matter of historical fact, the main aim of the "procedural studies" carried out in Geneva was to introduce concrete microgenesis into the macrogenetic Piagetian model. PMID- 11016553 TI - Behavioral and physiological responsivity, sleep, and patterns of daily cortisol production in infants with and without colic. AB - To describe the behavioral and physiological responses associated with colic, the responses of 20 two-month-old infants with and 20 without colic were studied during a physical examination. Parents kept a diary of infant behaviors (including crying and fussing) for 3 days following the visit. Using Wessel, Cobb, Jackson, Harris, & Detwiler criteria, colic was defined as fussing/crying for 3 hr or more on each of the 3 days. Behavioral data coded by "blind" observers showed that during the physical exam, colic infants cried twice as much, cried more intensely, and were more inconsolable than were control infants. Despite these behavioral differences, heart rate, vagal tone, and cortisol measures indicated no appreciable difference in physiological responsivity for the two groups. At home, parents collected saliva cortisol samples at wakeup, midmorning, midafternoon, and evening for 2 days. In a finding similar to that shown by the laboratory data, the colic and control infants did not have different levels of daily average cortisol. These laboratory and home data provide no evidence of greater responsivity in the physiological substrate of difficult temperament for colic infants and are consistent with evidence of similarity in temperament once colic is resolved. At home, compared with control infants, colic infants did display a blunted rhythm in cortisol production. By diary, they also slept about 2 hr less per day than did control infants. Nighttime sleep was still significantly different when fussing/crying was statistically controlled. These data suggest that colic might be associated with a disruption or delay in the establishment of the circadian rhythm in activity of the hypothalamic-pituitary-adrenocortical axis and associated sleep-wake activity. PMID- 11016554 TI - A study of multimodal motherese: the role of temporal synchrony between verbal labels and gestures. AB - This study examined European American and Hispanic American mothers' multimodal communication to their infants (N = 24). The infants were from three age groups representing three levels of lexical-mapping development: prelexical (5 to 8 months), early-lexical (9 to 17 months), and advanced-lexical (21 to 30 months). Mothers taught their infants four target (novel) words by using distinct objects during a semistructured play episode. Recent research suggests that young infants rely on temporal synchrony to learn syllable-object relations, but later, the role of synchrony diminishes. Thus, mothers' target and nontarget naming were coded for synchrony and other communication styles. The results indicated that mothers used target words more often than nontarget words in synchrony with object motion and sometimes touch. Thus, "multimodal motherese" likely highlights target word-referent relations for infants. Further, mothers tailored their communication to infants' level of lexical-mapping development. Mothers of prelexical infants used target words in synchrony with object motion more often than mothers of early- and advanced-lexical infants. Mothers' decreasing use of synchrony across age parallels infants' decreasing reliance on synchrony, suggesting a dynamical and reciprocal environment-organismic relation. PMID- 11016555 TI - Young children's understanding of perception, desire, and emotion. AB - Very young children seem to know that people experience several mental states: desires, perceptions, emotions. In three studies, we investigated 2- and young 3 year-olds' judgments and communications about how these states connect together in people's lives and minds. Two experimental studies with 56 participants demonstrated young children's understanding of at least one set of connections: In appropriate circumstances, a person's perception of desirable or undesirable objects leads to related emotional experiences. A complementary investigation of four young children's everyday conversations demonstrated their awareness of and expression of several additional connections between people's desires, perceptions, and emotions. PMID- 11016556 TI - The development of children's knowledge of the times of future events. AB - Four studies with 261 children were conducted to describe 4- through 10-year olds' ability to differentiate the future distances of events. Distances ranged from later the same day through nearly a year in the future. Judgment methods included pointing to parts of a spatial scale representing future distances and answering open-ended questions. Although 4-year-olds failed to differentiate future distances, 5-year-olds were able to distinguish events that would occur in the coming weeks and months from those that would not occur for many months. However, like young children in earlier studies of memory for time, they confused the near future with the recent past. Children 6 through 8 years of age made more differentiated judgments but collapsed the distances of events more than a few months in the future. By 8 to 10 years of age, children accurately judged distances by using mental representations of the times of events in the annual cycle. PMID- 11016557 TI - Children's memories for painful cancer treatment procedures: implications for distress. AB - Children (ages 3 to 18, N = 55) diagnosed with leukemia were tested for their memories of lumbar punctures (LPs), a repeated and painful part of the cancer treatment protocol. Memory for both event details and the child's emotional responses was assessed one week after the LP. Children of all ages displayed considerable accuracy for event details, and accuracy increased with age. Overall recall accuracy for event details and emotional responses was similar. Recall among children given oral Versed was similar to that among children not given Versed. Finally, higher distress predicted greater exaggerations in negative memory 1 week later (although controlling for age weakened this relationship); moreover, greater exaggerations in negative memory predicted higher distress at a subsequent LP. These results indicate that children's memories play an important role in their experience of distress during repeated stressful events. PMID- 11016558 TI - Genetic and environmental covariation between verbal and nonverbal cognitive development in infancy. AB - Despite cognitive neuroscience's emphasis on the modularity of cognitive processes, multivariate genetic research indicates that the same genetic factors largely affect diverse cognitive abilities, at least from middle childhood onward. We explored this issue for verbal and nonverbal cognitive development in infancy in a study of 1,937 pairs of same-sex 2-year-old twins born in England and Wales in 1994. The twins were assessed by having their parents use a measure of productive vocabulary (the MacArthur Communicative Development Inventory) and a novel measure of nonverbal cognitive abilities (Parent Report of Children's Ability). Verbal and nonverbal development correlated .42. A multivariate genetic analysis indicated that genetic factors were responsible for less than half of this phenotypic correlation. Moreover, the genetic correlation between verbal and nonverbal abilities was only .30, which indicates that genetic effects on verbal and nonverbal abilities are largely independent in infancy. These multivariate genetic results suggest that genetic effects on cognitive abilities are modular early in development and then become increasingly molar. The implications of this result for theories of cognitive development are discussed. PMID- 11016559 TI - The relation of child care to cognitive and language development. National Institute of Child Health and Human Development Early Child Care Research Network. AB - Children from 10 sites in the United States were followed from birth to age 3 to determine how experiences in child care relate to cognitive and language development (Ns varied between 595 and 856, depending on the assessment). Multiple assessments of family and child care environments and of cognitive and language competence were collected. Analyses that adjusted for maternal vocabulary score, family income, child gender, observed quality of the home environment, and observed maternal cognitive stimulation indicated that the overall quality of child care, and language stimulation in particular, was consistently but modestly related to cognitive and language outcomes at ages 15, 24, and 36 months. The effect sizes for high (top quartile) versus low (bottom quartile) quality ranged from .18 to .48. After adjusting for child care quality, cumulative experience in center-based care was associated with better outcomes than was participation in other types of care. The amount of time children spent in care was not related to outcomes. Children in exclusive maternal care did not differ systematically from children in child care. Tests for lagged relations of earlier child care experiences to later performance (adjusting for current child care) showed that language stimulation predicted subsequent cognitive and language performance 9 to 12 months later. Although children in center care at age 3 performed better than children in other types of care, earlier experience in child care homes was associated with better performance at age 3 than was experience in other types of care. The relations of child care variables to outcomes did not vary consistently as a function of family income, quality of home environment, child gender, or ethnic group. PMID- 11016560 TI - Developmental differences in visual and auditory processing of complex sentences. AB - Children aged 8 through 11 (N = 250) were given a word-by-word sentence task in both the visual and auditory modes. The sentences included an object relative clause, a subject relative clause, or a conjoined verb phrase. Each sentence was followed by a true-false question, testing the subject of either the first or second verb. Participants were also given two memory span measures: digit span and reading span. High digit span children slowed down more at the transition from the main to the relative clause than did the low digit span children. The findings suggest the presence of a U-shaped learning pattern for on-line processing of restrictive relative clauses. Off-line accuracy scores showed different patterns for good comprehenders and poor comprehenders. Poor comprehenders answered the second verb questions at levels that were consistently below chance. Their answers were based on an incorrect local attachment strategy that treated the second noun as the subject of the second verb. For example, they often answered yes to the question "The girl chases the policeman" after the object relative sentence "The boy that the girl sees chases the policeman." Interestingly, low memory span poor comprehenders used the local attachment strategy less consistently than high memory span poor comprehenders, and all poor comprehenders used this strategy less consistently for harder than for easier sentences. PMID- 11016561 TI - Memory strategy development: do we need yet another deficiency? AB - Recent discussions of strategy development have included the introduction of the concept of utilization deficiency. The present analysis examines the definition of utilization deficiency vis-a-vis the older contrasts between mediation and production deficiency and assesses the logical clarity of the current definition. Further, because utilization deficiency focuses on the transition from initial to proficient strategy use, the present analysis considers all of the possible types of strategy inefficiencies and evaluates whether the current definition of utilization deficiency precludes consideration of important strategy inefficiencies that have been documented in the existing literature and are likely to form the bulk of yet-to-be discovered inefficiencies. Although the emphasis on strategy inefficiencies is welcomed, the current analysis concludes that there are serious problems with the current definition of utilization deficiency, problems that both obscure important theoretical distinctions of the past and limit the investigation of strategy inefficiencies that are likely to play an important role in our understanding of the development of strategy use. Furthermore, the linear developmental model that frames utilization deficiencies from no benefit to sophisticated strategy use ignores the heterogeneity in strategy development that has been recently documented. PMID- 11016562 TI - How best to utilize a deficiency. AB - This article is a commentary on Harriet Waters' discussion, in this issue, of strategy inefficiencies and strategy utilization, mediation, and production deficiencies. The author focuses on the importance and meaning of the degree of spontaneity in strategy production, and situates the concept of utilization deficiency within current work on strategy heterogeneity, contextual support, and situation-specific skills. PMID- 11016563 TI - Child compliance/noncompliance and maternal contributors to internalization in maltreating and nonmaltreating dyads. AB - Moral development in maltreated and nonmaltreated children was examined by coding child compliance and noncompliance behaviors in a mother-child interaction during a cleanup situation that followed a semistructured free play. Features of child compliance/noncompliance involve a shift from reliance on external controls to internal mechanisms, thereby reflecting child internalization of the maternal agenda. Differences in maltreating versus comparison mothers' use of control strategies (power-assertive and inductive techniques) and their relations to child internalization were examined. Eighty-nine mother-child dyads participated; approximately half of the children (n = 46) had documented histories of maltreatment and the remaining children (n = 43) were nonmaltreated, demographically similar comparison children. Maltreated children were divided into two subgroups: physically abused and neglected. Compared with nonmaltreated children, abused children were found to exhibit less internalization, whereas neglected children displayed significantly more negative affect. No differences were found between groups for the maternal control strategies. However, maltreated and nonmaltreated groups differed in the maternal variables that predicted child internalization. A lower level of maternal negative affect was linked to child internalization in maltreated children, whereas a lower level of maternal joy predicted internalization for the comparison children. The findings suggest that maltreated children exhibit both behavioral and affective differences in their moral development, with differential effects based on the type of maltreatment. The clinical implications for maltreated children's self and moral development are discussed. PMID- 11016564 TI - Parent and peer contexts for children's moral reasoning development. AB - This study addressed the polarization among theoretical perspectives in moral psychology regarding the relative significance of parents and peers in children's developing moral maturity. The sample was composed of 60 target children from late childhood and midadolescence, 60 parents, and 60 friends who participated in parent/child and friend/child dyadic discussions of a series of moral conflicts. The quality of parents' and friends' verbal interactions, ego functioning, and level of moral reasoning in these discussions was used to predict the rate of children's moral reasoning development over a 4-year longitudinal interval. Results revealed that interactions with both parents and peers were predictive of children's development but that these two types of relationships influence development in rather different ways. Implications of the findings for the understanding of these socialization agents' roles in moral development are discussed. PMID- 11016565 TI - Adolescent social competence: effectiveness in a group setting. AB - We used a longitudinal data set to evaluate an observational assessment of adolescent competence within a group context. Participants were 40 (21 males, 19 females) ethnically diverse children who had earlier been observed in summer camp (age 10) and preschool (age 4 1/2), as well as in infancy. A revealed differences task was videotaped as part of an intensive weekend camp reunion at ages 15 to 16. Raters, blind to the adolescents' developmental histories, coded each adolescent on the following behavioral rating scales: enjoyment of the task, involvement, leadership, self-confidence in the task, and global social competence. Both concurrent validity (strong correlations with independent camp reunion counselor ratings and peer sociometric measures) and clear associations with antecedent measures of peer competence in preschool and middle childhood were demonstrated. A pattern of correlations revealed considerable discriminate validity. Neither IQ nor socioeconomic status accounted for these associations. Overall, the results confirm the power of a developmentally appropriate, taxing, behaviorally based assessment of group functioning as a measure of competence in adolescence. PMID- 11016566 TI - Mastery motivation and self-evaluative affect in toddlers: longitudinal relations with maternal behavior. AB - This study examined relations between maternal control and evaluative feedback during the second year of life and children's mastery motivation and expressions of self-evaluative affect a year later. Participants were 75 toddlers (35 girls, 40 boys) and their mothers. Maternal controlling behavior and evaluative feedback were examined while mothers taught their 24-month-olds a challenging task. Children's mastery motivation and expressions of self-evaluative affect were assessed during easy and difficult achievement-like tasks when they were 36 months old. Maternal evaluative feedback and control style at 24 months predicted children's shame, persistence, and avoidance of mastery activities at 36 months. Specifically, negative maternal evaluations at age two related to children's later shame, especially when feedback was linked to children's actions or products; positive maternal feedback overall, as well as corrective feedback, related to children's later persistence; mothers who engaged in more autonomy supporting control with their 2-year-olds had children who were less likely to avoid challenging activities at age 3. Children's pride at 36 months was not predicted by mothers' behavior at 24 months. PMID- 11016567 TI - Parental monitoring: a reinterpretation. AB - Monitoring (tracking and surveillance) of children's behavior is considered an essential parenting skill. Numerous studies show that well-monitored youths are less involved in delinquency and other normbreaking behaviors, and scholars conclude that parents should track their children more carefully. This study questions that conclusion. We point out that monitoring measures typically assess parents' knowledge but not its source, and parents could get knowledge from their children's free disclosure of information as well as their own active surveillance efforts. In our study of 703 14-year-olds in central Sweden and their parents, parental knowledge came mainly from child disclosure, and child disclosure was the source of knowledge that was most closely linked to broad and narrow measures of delinquency (normbreaking and police contact). These results held for both children's and parents' reports, for both sexes, and were independent of whether the children were exhibiting problem behavior or not. We conclude that tracking and surveillance is not the best prescription for parental behavior and that a new prescription must rest on an understanding of the factors that determine child disclosure. PMID- 11016568 TI - The similarity of siblings' attachments to their mother. AB - Do siblings develop similar attachment relationships with their mother? Attachment theory suggests that brothers and sisters growing up in the same family are likely to relate in similar ways to their parents, at least when parental attachment representations and interactive styles remain stable across time. In the current study, sibling attachment data from three research groups (from Pennsylvania State University, Leiden University, and the University of Western Ontario) have been pooled to assemble a sufficiently large sample of observations (N = 138 sibling pairs) for a detailed comparison of sibling attachment relationships. Spacing between the births, differences in maternal sensitivity, and gender of siblings were examined as possible sources of concordance of nonconcordance. Attachment security (including disorganized attachment) of each sibling was assessed with the Strange Situation procedure between 12 and 14 months after birth. Maternal sensitivity was observed with the same rating scale in a laboratory play session in one of the studies and in home observations in the others. Sibling relationships were found to be significantly concordant when classified as secure/nonsecure (62% concordance, p < .01, 1 tailed, intraclass correlation = .23) but not when further subcategorized. Maternal insensitivity to both siblings (shared environment) was associated with concordance of sibling nonsecurity. Siblings of the same gender were more likely to form concordant relationships with their mother (68%; p < .01, 1-tailed, intraclass correlation = .37) than those of opposite gender. Same-sex sibling concordance was comparable to the concordance found for monozygotic twins in earlier studies. Genetic factors may, therefore, play a relatively small role in the development of attachment. PMID- 11016569 TI - Coping efficacy and psychological problems of children of divorce. AB - Three models of the relations of coping efficacy, coping, and psychological problems of children of divorce were investigated. A structural equation model using cross-sectional data of 356 nine- to twelve-year-old children of divorce yielded results that supported coping efficacy as a mediator of the relations between both active coping and avoiding coping and psychological problems. In a prospective longitudinal model with a subsample of 162 of these children, support was found for Time 2 coping efficacy as a mediator of the relations between Time 1 active coping and Time 2 internalizing of problems. Individual growth curve models over four waves also found support for coping efficacy as a mediator of the relations between active coping and psychological problems. No support was found for alternative models of coping as a mediator of the relations between efficacy and symptoms or for coping efficacy as a moderator of the relations between coping and symptoms. PMID- 11016570 TI - Striate cortex increases contrast gain of macaque LGN neurons. AB - Recurrent projections comprise a universal feature of cerebral organization. Here, we show that the corticofugal projections from the striate cortex (VI) to the lateral geniculate nucleus (LGN) robustly and multiplicatively enhance the responses of parvocellular neurons, stimulated by gratings restricted to the classical receptive field and modulated in luminance, by over two-fold in a contrast-independent manner at all but the lowest contrasts. In the equiluminant plane, wherein stimuli are modulated in chromaticity with luminance held constant, such enhancement is strongly contrast dependent. These projections also robustly enhance the responses of magnocellular neurons but contrast independently only at high contrasts. Thus, these results have broad functional significance at both network and neuronal levels by providing the experimental basis and quantitative constraints for a wide range of models on recurrent projections and the control of contrast gain. PMID- 11016572 TI - In search of the visual pigment template. AB - Absorbance spectra were recorded by microspectrophotometry from 39 different rod and cone types representing amphibians. reptiles, and fishes, with A1- or A2 based visual pigments and lambdamax ranging from 357 to 620 nm. The purpose was to investigate accuracy limits of putative universal templates for visual pigment absorbance spectra, and if possible to amend the templates to overcome the limitations. It was found that (1) the absorbance spectrum of frog rhodopsin extract very precisely parallels that of rod outer segments from the same individual, with only a slight hypsochromic shift in lambdamax, hence templates based on extracts are valid for absorbance in situ: (2) a template based on the bovine rhodopsin extract data of Partridge and De Grip (1991) describes the absorbance of amphibian rod outer segments excellently, contrary to recent electrophysiological results; (3) the lambdamax/lambda invariance of spectral shape fails for A1 pigments with small lambdamax and for A2 pigments with large lambdamax, but the deviations are systematic and can be readily incorporated into, for example, the Lamb (1995) template. We thus propose modified templates for the main "alpha-band" of A1 and A2 pigments and show that these describe both absorbance and spectral sensitivities of photoreceptors over the whole range of lambdamax. Subtraction of the alpha-band from the full absorbance spectrum leaves a "beta-band" described by a lambdamax-dependent Gaussian. We conclude that the idea of universal templates (one for A1- and one for A2-based visual pigments) remains valid and useful at the present level of accuracy of data on photoreceptor absorbance and sensitivity. The sum of our expressions for the alpha- and beta-band gives a good description for visual pigment spectra with lambdamax > 350 nm. PMID- 11016571 TI - Rapid identification of ocular dominance columns in macaques using cytochrome oxidase, Zif268, and dark-field microscopy. AB - Strabismus induces an abnormal pattern of alternating light and dark columns of cytochrome oxidase (CO) activity in macaque striate cortex. This pattern may arise because visual perception is suppressed in one eye to avoid diplopia. To test whether CO activity is reduced in the ocular dominance columns of the suppressed eye, we performed monocular enucleation to co-label the ocular dominance columns with Zif268 immunohistochemistry in seven exotropic adult Macaca fascicularis. This approach was unsuccessful, for two reasons. First, Zif268 yielded inconsistent labelling, that was usually greater in the enucleated eye's ocular dominance columns, but was sometimes greater in the intact eye's columns. Therefore, Zif268 was not a reliable method for identifying the ocular dominance columns serving each eye. Second, in three control animals we found that a brief survival period following monocular enucleation (needed for Zif268 levels to change) was long enough to alter CO staining. For example, a survival time of only 3 h was sufficient to induce CO columns, indicating that the activity of this enzyme fluctuates more rapidly than realized previously. Independent of these findings, we have also discovered that acute monocular enucleation produces a vivid pattern of ocular dominance columns visible in unstained or CO-stained sections under dark-field illumination. The ocular dominance columns of the acutely enucleated eye appear dark. This was verified by labelling the ocular dominance columns with [3H]proline. Dark-field illumination of the cortex after acute monocular enucleation offers a new, easy method for identifying the ocular dominance columns in macaques. PMID- 11016573 TI - Projections of the superior colliculus to subdivisions of the inferior pulvinar in New World and Old World monkeys. AB - Patterns of terminals labeled after WGA-HRP injections in the superior colliculus (SC) in squirrel monkeys and macaque monkeys, and after DiI application in marmosets, were related to the architecture of the pulvinar and dorsal lateral geniculate nucleus (LGN). In all studied species, the SC projects densely to two architectonic subdivisions of the inferior pulvinar, the posterior inferior pulvinar nucleus (PIp) and central medial inferior pulvinar nucleus (PIcM). These projection zones expressed substance P. Thus, sections processed for substance P reveal SC termination zones in the inferior pulvinar. The medial subdivision of the inferior pulvinar, PIm, which is known to project to visual area MT, does not receive a significant collicular input. Injections in MT of a squirrel monkey revealed no overlap between SC terminals and neurons projecting to area MT. Thus, PIm is not the significant relay station of visual input from the SC to MT. The SC also sends an input to the LGN, however, this projection is sparser than the input directed to pulvinar. PMID- 11016574 TI - Visual signals used in time-interval discrimination. AB - Thresholds for the detection of differences in the duration of visual stimuli were determined for a variety of programs of stimulus onset and offset. Performance suffers when a time interval begins with an ON step and ends with another ON stimulus, compared to the standard ON-OFF stimulation, but the decrement is reversed when the light is ramped down to background during the interval. Neither the magnocellular nor the parvocellular streams can be excluded because there is relatively little impairment of duration discrimination when the stimulus has low contrast or is heterochromatic at isoluminance. Performance at a variety of intensity levels suggests that sustained neural firing in an early stage of visual processing provides a background activity, which prevents good temporal precision of signals. PMID- 11016575 TI - Direct imaging of NMDA-stimulated nitric oxide production in the retina. AB - In the retina, nitric oxide (NO) functions in network coupling, light adaptation, neurotransmitter receptor function, and synaptic release. Neuronal nitric oxide synthase (nNOS) is present in the retina of every vertebrate species investigated. However, although nNOS can be found in every retinal cell type, little is known about the production of NO in specific cells or about the diffusion of NO within the retina. We used diaminofluorescein-2 (DAF-2) to image real-time NO production in turtle retina in response to stimulation with N-methyl D-aspartate (NMDA). In response to NMDA, NO was produced in somata in the ganglion cell and inner nuclear layers, in synaptic boutons and processes in the inner plexiform layer, in processes in the outer plexiform layer, and in photoreceptor inner segments. This NO-dependent fluorescence production quickly reached transient peaks and declined more slowly toward baseline levels at different rates in different cells. In some cases, the NO signal was primarily confined to within 10 microm of the source, which suggests that NO may not diffuse freely through the retina. Such limited spread was not predicted and suggests that NO signal transduction may be more selective than suggested, and that NO may play significant intracellular roles in cells that produce it. Because NO-dependent fluorescence within amacrine cells can be confined to the soma, specific dendritic sites, or both with distinct kinetics, NO may function at specific synapses, modulate gene expression, or coordinate events throughout the cell. PMID- 11016576 TI - Morphology of wide-field bistratified and diffuse human retinal ganglion cells. AB - To study the detailed morphology of human retinal ganglion cells, we used intracellular injection of horseradish peroxidase and Neurobiotin to label over 1,000 cells in an in vitro, wholemount preparation of the human retina. This study reports on the morphology of 119 wide-field bistratified and 42 diffuse ganglion cells. Cells were analyzed quantitatively on the basis of dendritic field size, soma size, and the extent of dendritic branching. Bistratified cells were similar in dendritic-field diameter (mean +/- S.D. = 682 +/- 130 microm) and soma diameter (mean +/- S.D. = 18 +/- 3.3 microm) but showed a broad distribution in the extent of dendritic branching (mean +/- S.D. branch point number = 67 +/- 32; range = 15-167). Differences in the extent of branching and in dendritic morphology and the pattern of branching suggest that the human retina may contain at least three types of wide-field bistratified cells. Diffuse ganglion cells comprised a largely homogeneous group whose dendrites ramified throughout the inner plexiform layer. The diffuse cells had similar dendritic-field diameters (mean +/- S.D. = 486 +/- 113 microm), soma diameters (mean +/- S.D. = 16 +/- 2.3 microm), and branch points numbers (mean +/- s.D. = 92 +/- 32). The majority had densely branched dendritic trees and thin, very spiny dendrites with many short, fine, twig-like thorny processes. Five of the diffuse cells had much more sparsely branched dendritic trees (<50 branch points) and less spiny dendrites, suggesting that there are possibly two types of diffuse ganglion cells in human retina. Although the presence of a diversity of large bistratified and diffuse ganglion cells has been observed in a variety of mammalian retinas, little is known about the number of cell types, their physiological properties, or their central projections. Some of the human wide-field bistratified cells in the present study, however, show morphological similarities to monkey large bistratified cells that are known to project to the superior colliculus. PMID- 11016577 TI - Visual evoked potentials and magnocellular and parvocellular segregation. AB - We have measured visual evoked potentials (VEPs) to luminance-modulated, square wave alternating, 3-deg homogeneous disks for stimulus frequencies ranging from 1 Hz to 16.7 Hz. The aim of the study was to determine the range of frequencies at which we could reproduce the two-branched contrast-response (C-R) curves we had seen at 1 Hz (Valberg & Rudvin, 1997) and which we interpreted as magnocellular (MC) and parvocellular (PC) segregation. Low-contrast stimuli elicited relatively simple responses to luminance increments resulting in waveforms that may be the signatures of inputs from magnocellular channels to the visual cortex. At all frequencies, the C-R curves of the main waveforms were characterized by a steep slope at low contrasts and a leveling off at 10%-20% Michelson contrast. This was typically followed by an abrupt increase in slope at higher contrasts, giving a distinctive two-branched C-R curve. On the assumption that the low-contrast, high gain branch reflects the responsivity of magnocellular-pathway inputs to the cortex, the high-contrast branch may be attributed to additional parvocellular activation. While a two-branched curve was maintained for frequencies up to 8 Hz, the high-contrast response was significantly compromised at 16.7 Hz, revealing a differential low-pass filtering. A model decomposing the measured VEP response into two separate C-R curves yielded a difference in sensitivity of the putative MC- and PC-mediated response that, when plotted as a function of frequency, followed a trend similar to that found for single cells. Due to temporal overlap of responses, the MC and PC contributions to the waveforms were hard to distinguish in the transient VEP. However, curves of time-to-peak (delay) as a function of contrast often went through a minimum before the high-contrast gain increase of the corresponding C-R curve, supporting the notion of a recruitment of new cell ensembles in the transition from low to high contrasts. PMID- 11016578 TI - The mosaic of horizontal cells in the macaque monkey retina: with a comment on biplexiform ganglion cells. AB - To further characterize the H1 and H2 horizontal cell populations in macaque monkey retinae, cells were injected with the tracer Neurobiotin following intracellular recordings. Tracer coupling between cells of the same type revealed all H1 or H2 cells in small patches around the injected cell. The mosaics of their cell bodies and the tiling of the retina with their dendrites were analyzed. Morphological differences between the H1 and H2 cells observable in Neurobiotin-labeled patches made it possible to recognize H1 and H2 cells in retinae immunolabeled for the calcium-binding proteins parvalbumin and calbindin, and thus to study their relative spatial densities across the retina. These data, together with the intracellularly stained patches, show that H1 cells outnumber H2 cells at all eccentricities. There is, however, a change in the relative proportions of H1 and H2 cells with eccentricity: close to the fovea the ratio of H1 to H2 cells is approximately 4 to 1, in midperipheral retina approximately 3 to 1, and in peripheral retina approximately 2 to 1. In both the Neurobiotin stained and the immunostained retinae, about 3-5% of the H2 cells were obviously misplaced into the ganglion cell layer. Several features of the morphology of the misplaced H2 cells suggest that they represent the so-called "biplexiform ganglion cells" previously described in Golgi studies of primate retina. PMID- 11016579 TI - Color-opponent responses of small and giant bipolar cells in the carp retina. AB - The physiological and morphological properties of color-opponent bipolar cells in the carp retina were studied. Fifty nine OFF-center bipolar cells and 63 ON center bipolar cells out of about 500 total bipolar cells recorded showed color opponent responses. The OFF-center color-opponent bipolar cells were classified into three subgroups according to their spectral and spatial responses. Fifty OFF center color-opponent cells responded with depolarization to a blue light spot and with hyperpolarization to a red spot in the receptive-field center. The polarity of the surround response was opposite to that of center response at each wavelength. Therefore these cells were classified as OFF double-opponent cells (OFF-DO). Eight cells responded with hyperpolarization to a blue and green spot and with depolarization to a red spot. The surround responses of those cells were depolarizing at any wavelength (R+G- cell). One responded with hyperpolarization to a blue and red spot and with depolarization to a green spot. The surround response showed a different spectral characteristic from that of the center response. It responded with depolarization to a blue and green annulus and with hyperpolarization to a red annulus (R-G+B- cell). The ON-center color-opponent bipolar cells were similarly classified into three subgroups. Sixty of ON-center color-opponent cells were the double color-opponent type (ON-DO cell), showing the responses of opposite polarity to the OFF-DO cells. Two cells were classified as R- G+ cell, and one cell as R+G-B+ cell. Both OFF- and ON-DO cells were identified by their morphology as Cajal's giant bipolar cells, and R+G-, R-G+, R G+B-, and R+G-B+ cells as Cajal's small bipolar cells. The analysis of the latency and the ionic mechanisms of their responses suggest that DO cells under light-adapted conditions receive direct inputs from long-wavelength (red) cones, RG cells from middle-wavelength (green) cones, and RGB cells from short wavelength (blue) cones. Possible mechanisms of the opponent inputs to these bipolar cells are discussed. PMID- 11016581 TI - The sequential processing of visual motion in the human electroretinogram and visual evoked potential. AB - Mechanisms of motion vision in the human have been studied extensively by psychophysical methods but less frequently by electrophysiological techniques. It is the purpose of the present investigation to study electrical potentials of the eye (electroretinogram, ERG) and of the brain (visual evoked potential, VEP) in response to moving regular square-wave stripe patterns spanning a wide range of contrasts, spatial frequencies, and speeds. The results show that ERG amplitudes increase linearly with contrast while VEPs, in agreement with the literature, show an amplitude saturation at low contrast. Furthermore, retinal responses oscillate with the fundamental temporal stimulus frequency of the moving pattern while brain responses do not. In both the retina and the brain, the response amplitudes are tuned to certain speeds which is in agreement with the nonlinear correlation-type motion detector. Along the ascending slopes (which means increasing amplitudes) of the tuning functions, the ERG curves overlap at all spatial frequencies if plotted as a function of temporal stimulation frequency. The ascending slopes of the tuning functions of the VEP overlap if plotted as a function of speed. The descending slopes (which means decreasing amplitudes) of the tuning functions show little (ERG) or no (VEP) overlap and the waveforms at high speeds approach pattern-offset-onset responses. These observations suggest that in the retina motion processing along the ascending slopes of the tuning curves takes place by coding the temporal stimulation frequency which depends on the spatial frequency of the moving pattern. In the brain, however, motion processing is by speed independent of spatial frequency. Simple calculations show that the VEP information is decoded from the ERG signal into a speed signal. PMID- 11016580 TI - Morphology and visual pigment content of photoreceptors from injured goldfish retina. AB - Adult teleost fish retinas can regenerate neurons following either surgical or pharmacological injury. The cellular milieu of the damaged retina within which regenerated neurons are produced might be different in these two model systems of retinal injury, and thus the phenotypic attributes of regenerated neurons in the two model systems might also differ. To determine if the phenotypic attributes of photoreceptors, and by extension the recovery of vision, are different between these two model systems, we compared the visual pigment content and morphology of photoreceptors derived from goldfish retinas of both models with control retina. Visual pigments-which consist of a protein moiety (opsin) and a chromophore--were analyzed in single, isolated photoreceptors using microspectrophotometric techniques. We report that visual pigments and photoreceptor morphologies in the surgical model closely matched those of native retina. In contrast, neither photoreceptor morphology nor visual pigment content matched closely in the pharmacological model. The results indicate that phenotypic attributes of photoreceptors can differ significantly between the two model systems of retinal regeneration, but that in both systems, rod- and cone-mediated visual functions can potentially be reestablished. PMID- 11016582 TI - Voltage-gated Na+ channel EOIII-segment-like immunoreactivity in fish retinal ganglion cells. AB - Although single-channel and whole-cell patch-clamp recordings have demonstrated the presence of Na+ currents in retinal ganglion cell somata, it has not previously been reported that an anti-Na+-channel antiserum stains both retinal ganglion cell somata and proteins with molecular weights corresponding to complexes of alpha and beta subunits. We probed adult goldfish retinas for Na+ channel-like immunoreactivity with a polyclonal antibody directed against the EOIII segment of vertebrate voltage-gated Na+ channels. In vertical sections and whole mounts, this antibody consistently stained the somata, axons, and proximal dendrites of retinal ganglion cells. Some somata in the proximal third of the inner nuclear layer were also stained. In Western blots, this antibody specifically stained multiple protein bands from retina and optic nerve, all with apparent molecular weights between 200 and 315 kDa. The largest of these molecular weights agrees with that reported previously for complexes of alpha and beta subunits in mammalian neurons, including retinal ganglion cells. The intermediate and lowest molecular weights are consistent with the presence of multiple Na+ channel alpha subunits, either in individual proximal retinal neurons or in different morphological subtypes. PMID- 11016583 TI - Survivability and immune responses after challenge with Edwardsiella ictaluri in susceptible and resistant families of channel catfish, Ictalurus punctatus. AB - Diseases in catfish farming are prevalent and costly, particularly the bacterial disease Enteric Septicemia of Catfish. Considerable research has focused on different aspects of this disease, including the biology of the causative agent, Edwardsiella ictaluri. However, no satisfactory treatment or preventive has resulted from these efforts. One solution is to increase the natural disease resistance of the fish through genetic selection. Recent research has demonstrated that genetic factors influence resistance to infection in mammals as well as fish. Selective breeding for disease resistance in channel catfish is ongoing, however differences in defence mechanisms among E. ictaluri challenged strains and families are only now being investigated. Antigen-specific as well as non-specific immune responses of full-sib families of channel catfish to laboratory challenge with E. ictaluri have been investigated. Both resistant and sensitive families produce a humoral response as specific antibody, but there were no differences found in the level of specific antibody produced. The sensitive family produced a slightly higher percentage of B lymphocytes in mononuclear cell preparations from peripheral blood, while the resistant family had a higher percentage of T lymphocytes in those preparations. The most significant observation was that the resistant family produced more macrophage aggregations in the spleen and posterior kidney throughout the infection than the sensitive family. Neither family produced stress-associated amounts of cortisol. PMID- 11016584 TI - Isolation and characterisation of a serum lectin from blue gourami, Trichogaster trichopterus (Pallus). AB - A novel lectin, designated BGL, has been purified from the serum of blue gourami, Trichogaster trichopterus, with the use of (NH4)2SO4 fractionation, affinity chromatography and gel filtration chromatography. Electrophoretic analyses and mass spectrometric study of purified BGL showed that the lectin is composed of two isoforms with native molecular masses estimated to be 65 and 66 kDa, and two subunits of 32 and 34 kDa on SDS-PAGE under non-reducing conditions. Upon reduction with 20 mM dithiothreitol (DTT), BGL showed two close bands of 27 and 29 kDa. After isoelectric focusing, the lectin focused as close double bands at pH 5.6. The N-termini of both isoforms share the same sequence (HGEENRXGPR) and show no significant homology with any known proteins. The BGL agglutinating activity is specifically inhibited by N-acetyl-D-galactosamine and N-acetyl-D glucosamine, and to a lesser degree by D-(+)-mannose, but not by D-(+)-galactose, D-(+)-glucose, maltose or N-acetyl-D-mannosamine. Haemagglutination assay showed that BGL is more specific for rabbit than mouse, chicken, rat or guinea pig erythrocytes, and haemagglutination was Ca2+-dependent. In addition, BGL could agglutinate a range of micro-organisms and yeast cells, with the exception of some fish pathogens, such as Aeromonas hydrophila (strains: PPD 134/91 and PPD 11/90) and Vibrio harveyi (strain: W618). Localisation of BGL by fluorescein isothiocyanate (FITC)-labelled antibodies revealed that the lectin is associated with the cell surface of fish leukocytes. PMID- 11016585 TI - Immunomodulation by dietary beta-1, 3-glucan in the brooders of the black tiger shrimp Penaeus monodon. AB - The present study evaluated the effectiveness of beta-1,3-glucan derived from Schizophyllum commune in enhancing shrimp survival as well as haemocyte phagocytosis and superoxide anion production in brooder Penaeus monodon. Pond reared P. monodon adults (135 +/- 25 g) stocked in outdoor or indoor tanks were fed either a test diet containing beta-1,3-glucan (2.0 g kg(-1) or a glucan-free control diet for 40 days. Their survival was compared. The brooders reared in indoor tanks were analysed at days 0, 1, 3, 6, 12, 24, 30 and 40 for their haemocyte phagocytic activity and superoxide anion production. The results showed that regardless of indoor or outdoor rearing the survival rate of shrimp fed the glucan diet was significantly higher (P<0.001) than that of the control group. The brooders showed enhanced haemocyte phagocytic activity, cell adhesion and superoxide anion production when glucan was administered in their diets. The immunostimulatory enhancement peaked at day 24 after starting the dietary exposure and subsequently decreased to the pre-feeding level at the end of the 40 days feeding trial. PMID- 11016586 TI - Characterisation of different morphological features of black tiger shrimp (Penaeus monodon) haemocytes using monoclonal antibodies. AB - Monoclonal antibodies (mabs) specific for Penaeus monodon haemocytes were produced by immunising mice with membrane lysates of shrimp haemocytes. Four mabs (WSH 6, WSH 7, WSH 8 and WSH 16) were characterised using flow cytometry, light microscopy, laser scanning microscopy, electron microscopy and immunoprecipitation. WSH 6 recognised a carbohydrate determinant on an 85 kDa molecule. WSH 7, WSH 8 and WSH 16 recognised 50, 35 and 115 kDa molecules, respectively. For all mabs, differences in amount and intensity of the labelling were found when haemocytes were fixed immediately in 2% formaldehyde in Alsever's Solution (AS), compared with non-fixed haemocytes that were kept in AS (which reduced activation of the haemocytes) or in L15 cell culture medium. WSH 6 reacted with the cell membranes of all fixed haemocytes, while WSH 7 and WSH 16 reacted with the cell membranes of >80% of fixed haemocytes. The membrane labelling appeared to decrease when cells were kept in L15 medium. WSH 8 did not react with the haemocyte membranes. All mabs reacted with some granules, mainly present in the hyaline cells, when the haemocytes were immediately fixed. When non-fixed cells were kept in AS and in L15 medium, positive granules were also observed in semigranular and granular haemocytes as well as in the largest granules of a fourth cell type, that contains many granules of different size and electron density. Immunoreactive extracellular thread-like material could be observed in cells in L15 medium. The change in staining pattern was extreme for WSH 8, somewhat less for WSH 6 and WSH 7 and the lowest for WSH 16. Double labelling revealed that all mabs showed a different staining pattern on membranes as well as on granules. WSH 16 also showed labelling in cytoplasmic vesicles, as well as in haemolymph plasma on histological sections. The hypothesis is put forward that immunoreactive molecules recognised by these mabs, are related to haemocyte activation factors. PMID- 11016587 TI - Inhibitory effects of morphine on some inflammation-related parameters in the goldfish Carassius auratus L. AB - Acute peritonitis induced in the goldfish by intraperitoneal injection of a sterile Thioglycollate solution shows a typical pattern with intraperitoneal exudation of serum proteins followed by influx of leucocytes (mainly heterophils/macrophages) correlated with elevated levels of chemotactic factors in peritoneal fluid and blood plasma. Supplementation of Thioglycollate with morphine (20 mg kg(-1) b.w.) does not affect the leakage of serum proteins into peritoneum. In contrast, it reduces the number of exudate peritoneal leucocytes (among them heterophils/macrophages) to the control level and decreases the level of peritoneal fluid/plasma chemoattractants, both effects being reversed by naltrexone pretreatment. Morphine itself acts as a chemokinetic factor for fish leucocytes as it increases their random movements. Therefore inhibitory effects of morphine on accumulation of exudate cells might be explained by inhibition of the production/release of chemotactic factors and/or reduced sensitivity of leucocytes to chemotactic signals. The effects of morphine on the goldfish peritonitis are in concordance with those described recently in Atlantic salmon and CB6 mice. PMID- 11016588 TI - Effects of injecting chitin particles on the innate immune response of gilthead seabream (Sparus aurata L.). AB - To determine the effects of chitin (poly [1-->4]-beta-N-acetyl-D--glucosamine) particles on the innate immune response of gilthead seabream (Sparus aurata L.), specimens were injected intravenously or intraperitoneally with the substance. Natural haemolytic complement activity, head-kidney leucocyte respiratory burst activity and phagocytic and cytotoxic activities were analysed in vitro 3, 5 or 10 days post-injection. All the immune parameters assayed remained unaffected when chitin was intravenously administered. However, the fish that had been intraperitoneally injected showed increased humoral and cellular immune responses. Natural haemolytic complement activity increased from 5 days post injection although no statistically significant differences were observed. Respiratory burst and phagocytic activities peaked at 3 and 5 days post injection, respectively, while cytotoxic activity had increased by 3 days post injection and remained high until 10 days post-injection. PMID- 11016589 TI - Enhancement of disease resistance against penaeid acute viraemia and induction of virus-inactivating activity in haemolymph of kuruma shrimp, Penaeus japonicus, by oral administration of Pantoea agglomerans lipopolysaccharide (LPS). PMID- 11016590 TI - Application of bacterins and yeast Acremonium dyosporii to protect the larvae of Macrobrachium rosenbergii from vibriosis. PMID- 11016591 TI - Viruses and multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system whose cause is still unknown. Many findings suggest an infectious etiology or, at least, that infectious agents in association with host genetic factors may play a role in the pathogenesis of this disease. Accumulating data, including animal models, human models of virus induced demyelination, epidemiologic, and laboratory findings, demonstrate that viruses and host genetic factors can interact to cause immune-mediated demyelination. While many viruses have been postulated as a possible cause of MS, to date, no "MS virus" has been definitively shown to be associated with this disease. Alternatively, ubiquitous viruses are being considered as the environmental "triggers" that have been postulated to be involved in the MS disease process. We will focus on recent studies with human herpesvirus 6 and MS as how a common virus may be associated with this disorder in a subset of infected individuals. PMID- 11016592 TI - Cell biology of virus entry, replication, and pathogenesis. AB - The Keystone Symposium on Cell Biology of Virus Entry, Replication, and Pathogenesis held in Taos, New Mexico February 29 to March 5, 2000 to a large degree fulfilled its purpose to facilitate interdisciplinary exchange of information. The Keynote Address by Sir John J. Skehel (National Institute for Medical Research, London) initiated discussion on the antigenic analyses of influenza virus hemagglutinins. Subsequent topics covered the earliest events in viral receptor-mediated entry through membrane fusion to virus assembly, genetics, pathogenesis, and immunology. While a significant number of taxonomically different viruses was discussed, several common themes emerged from plenary sessions and poster workshops. A few selected topics are highlighted below. PMID- 11016593 TI - Immunopathologic weight loss in intracranial LCMV infection initiated by the anorexigenic effects of IL-1beta. AB - Lymphocytic choriomeningitis virus (LCMV) infection of beta2-microglobulin deficient (beta2m-/-) mice results in a substantial loss of body weight that is not mediated by the virus itself, but rather by CD4+ T cells responding to the viral infection. In this study, we further characterized LCMV-induced weight loss in immunocompetent and beta32m-/- mice. We show that intracranial (i.c.), but not intraperitoneal (i.p.) LCMV infection elicited significant weight loss and that weight loss was preceded by anorexia. Also, uninfected mice fed an equivalent amount as eaten by infected mice had similar weight loss compared to their infected counterparts. Interestingly, both weight loss and anorexia were greater in female than male beta2m-/- mice. LCMV-infected female beta2m-/- mice also had significantly more interleukin (IL)-betag in their cerebrospinal fluid (CSF) than did male beta2m-/- mice. Finally, intracerebroventricular (i.c.v.) administration of anti-IL-1beta antibody, but not control immunoglobulin G (IgG), attenuated the initial weight loss and increased food intake. Taken together, these results suggest that the majority of weight loss after intracranial LCMV infection is the result of anorexia and IL-1beta mediates initial anorexic weight loss. PMID- 11016594 TI - Multifactorial protective mechanisms to limit viral replication in the lung of mice during primary murine cytomegalovirus infection. AB - In this article, we investigated the protective host immune mechanisms against acute murine cytomegalovirus (MCMV) infection. For this purpose, we used various knockout mice lacking molecules, which include interferon-gamma (IFN-gamma), interferon-gamma receptor (IFN-gamma-R), interferon regulatory factor-1 (IRF-1), inducible nitric oxide synthase (iNOS), and perforin. We also used mutant mice lacking Fas molecule. When we infected these mice with MCMV and determined the viral titers in their lungs at different time points, we found that IFN-gamma, IFN-gamma-R, IRF-1, iNOS, and perforin-deficient mice developed significantly higher titers of infectious MCMV in the lung, compared to those observed in their respective wild-type controls. In the lungs of Fas-mutant mice, viral titers were similar to those obtained in wild-type mice. PMID- 11016595 TI - Alleviation of renal disease and lymphadenopathy in MRL-Fasp(lrcg)/Fas(lprcg) (MR lpr(cg)) mice neonatally infected with mouse mammary tumor virus encoding superantigen strongly reactive with TCR Vbeta8.2 element. AB - Mouse mammary tumor virus transmitted by FM mice (FM-MMTV) encodes a superantigen (SAg) characterized by strong reactivity with TCR Vbeta8.2 element and broad spectrum of Vbeta reactivity. To investigate what effects the expression in vivo of FM-MMTV SAg exhibits on the course of the disease in a lupus-prone model, MRL/MpJ-Fas(lprcg)/Fas(lprcg) (MRL-lpr9cg) mice, neonatally FM-MMTV-infected MRL lprcg(MMTV) and uninfected MRL-lpr(cg) mice were compared for various disease parameters. In MRL-lprcg(MMTV), survival was significantly prolonged, glomerulonephritis, proteinuria, and lymphadenopathy were clearly ameliorated, and the production of serum immunoglobulin G (IgG), complement-activating IgG2a, and cryogenic IgG3 autoantibodies, which are thought to be pathogenic to kidneys, and circulating immune complexes (IC), and glomerular IC deposition were significantly suppressed. FM-MMTV infection deleted Vbeta8.2+ cells by about 90% and Vbeta14+ cells less efficiently in all of the CD4+, CD8+, and B220+ CD4- CD8- or double-negative (DN) T-cell populations, and Vbeta8.1+ cells in the CD4+ population but not in the others. Similar deletion profiles of CD8+ and DN T cells support that DN T cells are derived from the CD8 lineage. The results imply that the specific regulation of the immune system with viral SAg has a potential for development of an attractive immunomodulatory therapy of autoimmune diseases. PMID- 11016596 TI - Immunophenotypic study of atypical lymphocytes. Generated in peripheral blood and spleen of nude mice After MHV-72 infection. AB - Inbred athymic nude mice (BALB/c) were injected subcutaneously with the wild-type murine gammaherpesvirus 72 (MHV-72), which has been shown to induce the infectious mononucleosis (IM)-like syndrome in immunocompetent mice. The mice were also injected with UV-irradiated MHV-72. We studied the pattern of acute and chronic infection in the blood cells of the nude mice and detected viral DNA sequences in the infected leukocytes by polymerase chain reaction (PCR) technique up to when the animal died, close to 1 month postinfection. Using the UV irradiated virus that induces an increase in mouse survival time, the viral sequences were present in the blood up to 3 months postinfection, then disappeared. We detected atypical lymphocytes in the blood of mice infected with both wt and UV-irradiated viruses. These atypical cells were similar in shape to those present in the blood of patients with IM induced by Epstein-Barr virus (EBV). Via Unscheduled DNA Synthesis (UDS), DNA synthesis was demonstrated in the atypical cells whose phenotype is identical to that of B cells, as shown with a panel of monoclonal antibodies. By double immunofluorescence staining, using an hyperimmune anti-MHV-72 serum and an anti-IgG + IgM + IgA monoclonal antibody, we demonstrated that these atypical B cells express some viral antigens. Contrary to the immunocompetent mice, the nude mice did not develop splenomegaly after infection with wt virus, probably due to the lack of T cell subsets. However, we observed an increase of nude mice B cells in the spleen. The nude mice died 1 month postinfection showing a high frequency (40%) of atypical lymphoblast-like B cells in the blood; the increase in natural killer (NK) cell number was not detected after infection. Such findings suggest that NK cells probably did not play an important role in immune response to the MHV infection in nude mice. Finally, this mouse model could play an important role in antigammaherpesviral therapy of immunocompromised patients. PMID- 11016597 TI - Antibodies to the major linear neutralizing domains of cytomegalovirus glycoprotein B among natural seropositives and CMV subunit vaccine recipients. AB - The gB protein (gpUL55) of human cytomegalovirus (CMV) contains C-terminal (AD-1) and N-terminal (AD-2) linear immunodominant neutralizing domains. To measure antibodies to these epitopes, a modified protein (delta-gB) lacking heavily glycosylated intervening regions, the transmembrane domain, and the cytoplasmic domain, was expressed in recombinant baculovirus-infected cells. Eighty-six percent of 600 naturally CMV-seropositive individuals and 93% of 121 gB vaccine recipients had antibodies to delta-gB as detected by enzyme-linked immunosorbent assay (ELISA). The antibody level in vaccinees (median optical density [OD] = 1.73) exceeded that in natural seropositives (median OD = 0.94; p < .0001). Eleven percent of 95 natural seropositives and 7% of 120 gB vaccinees lacked A-gB antibodies but had neutralizing activity. Among subjects with delta-gB antibody, there were weak correlations between antibody level and neutralizing titer. These data suggest that antibodies to linear neutralizing gB domains are highly prevalent in naturally-infected individuals and regularly develop in gB vaccinees. However, for some individuals, discontinuous and/or linear epitopes not represented on delta-gB may be more important in the generation of neutralizing responses. PMID- 11016598 TI - Preparation and induction of immune responses by a DNA AIDS vaccine. AB - In an effort to evaluate the feasibility of developing a safe DNA vaccine for acquired immunodeficiency syndrome (AIDS), we have prepared a plasmid-based immunogen modeled after a naturally occurring noninfectious mutant of the simian immunodeficiency virus (SIV). The mutant SIV genome produces defective virus particles that are noninfectious in vitro and nonpathogenic in vivo in rhesus macaques. Analysis of the mutant genome revealed a 1.6 kb deletion that is in frame and spans integrase, vif, vpx, and most of vpr and results in a pol/vpr gene fusion. This deletion was introduced into the parental pathogenic molecular clone and the U3 region of the 5' LTR was replaced with a cytomegalovirus promoter to produce a candidate DNA vaccine, pIV. After transfection with this plasmid, SIV gag and envelope proteins are expressed and properly processed in vitro. When injected into rabbits, pIV elicited an antibody response to SIV gp130 envelope glycoprotein with titers reaching 1:2048, and a strong lymphoproliferative response to SIV gp130 and whole SIV. The potential to produce defective virus particles in vivo without integrating into the host genome should result in both a strong humoral and cellular immune response in rhesus macaques. In addition, this approach offers a safe alternative to live attenuated vaccines and DNA vaccines that are capable of integration. PMID- 11016599 TI - Immunoreactive domains and integrin-binding motifs in adenovirus penton base capsomer. AB - A panel of nine independent mouse monoclonal antibodies (MAbs) against penton base capsomers of subgenus C adenovirus serotypes 2 (Ad2) and 5 (Ad5) were isolated and characterized. Two of them (1D2 and 5A5), raised against Ad5 virion as the immunogen, bound to sodium dodecyl sulfate (SDS)-resistant and subgenus C specific epitopes that were not present in subgenus B Ad3 penton base. The 1D2 and 5A5 epitopes were mapped to two distinct regions that did not belong to the main variable region carrying the integrin-binding RGD motif at position 340. For the other seven MAbs, raised against recombinant Ad2 penton base protein (9S pentamers), the epitopes were sensitive to SDS-denaturation, but reacted with native Ad2, Ad5, and Ad3 penton base. The epitopes recognized by the nine MAbs and by polyclonal antipenton base antibodies defined three major immunoreactive regions. One (I) mapped to the N-terminal domain (residues 116-165); the other two regions were almost symmetrically disposed on both sides of the integrin binding RGD motif at position 340, within residues 248-270 (II), and within residues 368-427 (III) in the C-terminal domain. Region II overlapped the fiber binding site in penton base (residues 254-260). None of the MAbs showed any detectable virus neutralization effect, but they all slightly augmented the efficiency of Ad-mediated gene transfer. Although none of their epitopes included the RGD-340 tripeptide, substitutions of the arginine residue in the RGD motif abolished the reactivity of six individual and distant epitopes, suggesting a major conformational role for the RGD-containing domain. PMID- 11016600 TI - Detection of genetic diversity among bovine immunodeficiency virus population by single-strand conformation polymorphism analysis. AB - Serial virus specimens rescued from rabbits, experimentally infected with bovine immunodeficiency (BIV) strain R29, were monitored for changes in quasispecies population, using the single-strand conformation polymorphism (SSCP) analysis. The generation of characteristic SSCP patterns enables the rapid differentiation of BIV variants derived from the conserved part on the env region of the BIV genome, reducing the need for expensive and time-consuming direct sequencing analyses. Our results showed genetic polymorphism among a number of sampled BIV population in experimentally infected rabbits. At least three SSCP patterns (BIV quasispecies) were detected. The SSCP analysis allows for an easy, sensitive, and rapid screening of genetic variants of the virus and the assessment of variation at a number of tissue target sites. These variations may relate to cell-type targets and/or disease progression, and could be significant to our understanding of lentiviral pathogenesis. PMID- 11016601 TI - Immune components in porcine mammary secretions. AB - Immune components present in mammary secretions are reviewed. In swine, the histological structure of the placenta prevents in utero transfer of immunoglobulins and mammary secretions are the sole source of maternal antibody for the neonate. In addition to immunoglobulins, porcine mammary secretions contain significant numbers of maternal cells of various types that may contribute to neonatal immunity, including phagocytes (neutrophils and macrophages), lymphocytes (B and T cells), and epithelial cells. Immunomodulating and/or antimicrobial substances, including lactoferrin, lysozyme, lactoperoxidase, and cytokines, are also present in mammary secretions and may contribute to the protection of the neonate. While the role of immunoglobulins in mammary secretions is well understood, the contribution of cellular components and non-specific immune factors to neonatal immunity remains to be defined. PMID- 11016602 TI - Research on factors allowing a risk assessment of spore-forming pathogenic bacteria in cooked chilled foods containing vegetables: a FAIR collaborative project. AB - Vegetables are frequent ingredients of cooked chilled foods and are frequently contaminated with spore-forming bacteria (SFB). Therefore, risk assessment studies have been carried out, including the following: hazard identification and characterisation--from an extensive literature review and expertise of the participants, B. cereus and C. botulinum were identified as the main hazards; exposure assessment--consisting of determination of the prevalence of hazardous SFB in cooked chilled foods containing vegetables and in unprocessed vegetables, and identification of SFB representative of the bacterial community in cooked chilled foods containing vegetables, determination of heat-resistance parameters and factors affecting heat resistance of SFB, determination of the growth kinetics of SFB in vegetable substrate and of the influence of controlling factors, validation of previous work in complex food systems and by challenge testing and information about process and storage conditions of cooked chilled foods containing vegetables. The paper illustrates some original results obtained in the course of the project. The results and information collected from scientific literature or from the expertise of the participants are integrated into the microbial risk assessment, using both a Bayesian belief network approach and a process risk model approach, previously applied to other foodborne hazards. PMID- 11016603 TI - Analysis of the role of the Listeria monocytogenes F0F1 -AtPase operon in the acid tolerance response. AB - As little is known about the genes involved in the induction of an acid tolerance response in Listeria monocytogenes, the role of the F0F1-ATPase was analyzed as a consequence of its role in the acid tolerance of a number of other bacteria and its conserved nature. It was found that acid adapted cells treated with N,N' dicyclohexylcarbodiimide (DCCD) exhibited greatly enhanced sensitivity to low pH stress. Degenerate primers were designed to amplify and sequence a portion of the atpD gene. Subsequently, a PCR product from atpA to atpD was identified. While we were unable to create a deletion in the atpA gene, the plasmid pORI19 was inserted in a region between atpA and atpG to reduce, rather than eliminate, expression of the downstream genes. As expected this mutant displayed enhanced resistance to neomycin and exhibited slower growth than the wild type strain. This mutant could still induce an acid tolerance response and remained susceptible to DCCD treatment, but its relative acid sensitivity was difficult to assess as a consequence of its slow growth. PMID- 11016604 TI - Effects of combined exposure of micrococcus luteus to nisin and pulsed electric fields. AB - Death and injury following exposure of Micrococcus luteus to nisin and pulsed electric field (PEF) treatment were investigated in phosphate buffer (pH 6.8, sigma = 4.8 ms/cm at 20 degrees C). Four types of experiment were carried out, a single treatment with nisin (100 IU/ml at 20 degrees C for 2 h), a single PEF treatment, a PEF treatment followed by incubation with nisin (as before) and addition of nisin to the bacterial suspension prior to the PEF treatment. The application of nisin clearly enhanced the lethal effect of PEF treatment. The bactericidal effect of nisin reduced viable counts by 1.4 log10 units. Treatment with PEF (50 pulses at 33 kV/cm) resulted in a reduction of 2.4 log10 units. PEF treatment followed by nisin caused a reduction of 5.2 log10 units in comparison with a 4.9 log10 units reduction obtained with nisin followed by PEF. Injury of surviving cells was investigated using media with different concentrations of salt. Sublethally damaged cells of M. luteus could not be detected by this means, following PEF treatment. PMID- 11016605 TI - Permeability barrier of the gram-negative bacterial outer membrane with special reference to nisin. AB - The effect of nisin pretreatment on organic acid-induced permeability increase in strains of Escherichia coli, Pseudomonas aeruginosa, P. marginalis, and Salmonella enterica sv. Typhimurium was investigated, using assays based on the uptake of a fluorescent dye 1-N-phenylnaphthylamine (NPN) and on the bacterial susceptibility to detergent-induced bacteriolysis. The outer membrane of bacteria which had been pretreated with nisin was shown to be less stable against 1 mM EDTA, as indicated by their significantly higher NPN uptake levels as compared to untreated bacteria. Upon challenge with a tenfold lower concentration of EDTA (0.1 mM) some nisin-treated strains (Typhimurium, P. marginalis) exhibited, however, NPN uptake levels which were lower than those seen in control bacteria, suggesting that nisin had stabilized their outer membrane. Nisin pretreatment also decreased the NPN uptake induced by citric or lactic acid or both in E. coli, P. marginalis, and Typhimurium, whereas in P. aeruginosa the pretreatment resulted in increased NPN uptake in response to citric and lactic acid. These results suggest that, with the exception of P. aeruginosa, nisin could protect bacteria from the outer membrane-disrupting effect caused by the acids. P. aeruginosa was, however, shown to be protected against bacteriolysis induced by the detergents sodium dodecylsulfate and Triton X-100. With a pair of isogenic mutants of Typhimurium differing in their cell surface charge it was shown that the NPN uptake response to I mM EDTA of the abnormally cationic strain was not significantly affected by nisin, whereas in the normal anionic strain nisin strongly strengthened the uptake. Our hypothesis based on these findings is that the normally anionic cell surface of Gram-negative bacteria has a tendency to bind the cationic nisin. The binding of nisin to the surface does not proceed to the cytoplasmic membrane, but in the outer membrane the bound nisin actually stabilizes its structure through electrostatic interactions. With the exception of EDTA, the organic acids at pH 4 did not cause leakage of cell contents from Typhimurium, indicating that these acids do not permeabilize the outer membrane to an extent required for cytoplasmic pore formation by nisin. PMID- 11016606 TI - In vitro digestibility of bacillus fermented soya bean. AB - Bacillus fermented legume products include among others dawadawa and soumbala made from African locust bean, and natto and kinema made from soya bean. Bacillus subtilis is the dominant species involved in the fermentation. During Bacillus fermentation for 48 h of autoclaved soya bean the quantity of soluble and dialyzable matter increased from 22% and 6% up to 65% and 40%, respectively. Protein and carbohydrate degradation during fermentation of soya bean with several Bacillus spp. was investigated and appeared to be substantial during the first 18 h of fermentation resulting in the release of high levels of peptides and oligosaccharides. In vitro digestibility was increased from 29% up to 33-43% after Bacillus fermentation for 48 h. It was shown that Bacillus spp. were able to degrade soya bean macromolecules to a large extent resulting in water-soluble low molecular weight compounds. In vitro digestion of Bacillus fermented soya bean using gastrointestinal enzymes only slightly increased the amount of dialyzable matter, which clearly demonstrated the beneficial effect of Bacillus fermentation on food nutrient availability. PMID- 11016607 TI - Application of a systematic experimental procedure to develop a microbial model for rapid fish shelf life predictions. AB - A systematic experimental procedure for fish shelf-life modelling was used to develop a model for predicting the quality of fish in the chill chain. For this, the growth of the naturally occurring bacteria pseudomonads, Shewanella putrefaciens, Enterobacteriaceae, lactic acid bacteria and yeasts, on gilt-head seabream (Sparus aurata), was studied at temperatures from 0 to 15 degrees C. The results from the microbiological, organoleptical and chemical analysis conducted on naturally contaminated fish as well as on inoculated sterile fish blocks identified pseudomonads as a good spoilage index. Growth of pseudomonads was modelled as a function of storage temperature and correlated to organoleptical shelf life. To reduce the time required for the enumeration of the initial pseudomonads number, which is crucial information for shelf life prediction, a conductance assay was established. Compared with the conventional microbiological tests, this method gave results in one-fourth of the time. PMID- 11016608 TI - Occurrence of the vancomycin-resistant genes vanA, vanB, vanCl, vanC2 and vanC3 in Enterococcus strains isolated from poultry and pork. AB - It is suspected that the use of avoparcin as a feeding antibiotic for the fat stock contributes to development of cross-resistance against vancomycin and teicoplanin. After isolating enterococci strains from poultry and pork meat by cultivation on citrate azide Tween carbonate agar (CATC) and screening the vancomycin resistance on Columbia colistin nalidixic acid agar (CNA, supplemented with 5% sheepblood and 5 mg vancomycin/l) the polymerase chain reaction (PCR) was used for the detection of the vancomycin resistance genes vanA ('high level'), vanB ('moderate high level'), vanC1, vanC2 and vanC3 ('low level'). Out of 1643 E.-isolates from 115 poultry and 50 pork samples, 420 isolates could be identified as vancomycin resistant, 202 isolates of which carry the vanA, one isolate both the vanA and the vanC1, 38 isolates the vanC1, 14 isolates the vanC2, nine isolates both the vanC1 and the vanC3 gene and 156 isolates carry no gene. The vanB gene was not found in these isolates. Comparing vanA-positive food isolates with those from different human sources by means of the pulsed field gel electrophoresis (PFGE) it could clearly be demonstrated that they do not show homological fingerprints according to the source of origin. It is therefore unlikely that there is a close genetic relationship between isolates from animal foodstuff and humans. PMID- 11016609 TI - Changes in culturability and virulence of Salmonella typhimurium during long-term starvation under desiccating conditions. AB - The survival of Salmonella typhimurium under desiccation and starvation conditions commonly associated with farm buildings was investigated in a desiccation model system: filtration onto polycarbonate membranes placed in a sealed desiccator with 0.0067 g/m3 absolute humidity. Heterogeneities within bacterial populations in relation to time of desiccation were investigated on a single-cell basis by epifluorescence microscopy coupled with an image analysis system in conjunction with fluorescent dyes Chemchrome V6 and DAPI. Changes in cellular states were compared to the results of plate counts (colony forming units, CFU) on selective (modified semi-solid Rappaport Vassiliadis (MSRV)) and non-selective (nutrient agar (NA) and R2A agar) media, and to the measurements of infectivity and virulence using two animal models (chicks and mice). During 9 weeks of experimental desiccation, total cell counts (DAPI) of starved S. typhimurium remained stable, as did esterase activity (Chemchrome V6), but DAPI fluorescence intensity decreased slowly. Bacterial cells entered gradually into non-culturable states (decrease of CFU counts on MSRV, NA and R2A agar media) and the total loss of culturability on NA (defined as probability of presence of 1 CFU on the membrane inferior to 10 (-6)) was obtained after 9 weeks. Loss of chick infectivity and mice virulence in animal models occurred more rapidly, within three weeks of experimental desiccation. PMID- 11016610 TI - New developments in chromogenic and fluorogenic culture media. AB - This review describes some recent developments in chromogenic and fluorogenic culture media in microbiological diagnostic. The detection of beta-D glucuronidase (GUD) activity for enumeration of Escherichia coli is well known. E. coli O157:H7 strains are usually GUD-negative and do not ferment sorbitol. These characteristics are used in selective media for these organisms and new chromogenic media are available. Some of the new chromogenic media make the Salmonella diagnostic easier and faster. The use of chromogenic and fluorogenic substrates for detection of beta-D-glucosidase (beta-GLU) activity to differentiate enterococci has received considerable attention and new media are described. Rapid detection of Clostridium perfringens, Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus are other application of enzyme detection methods in food and water microbiology. PMID- 11016611 TI - Inhibition of fungal growth on bread by volatile components from spices and herbs, and the possible application in active packaging, with special emphasis on mustard essential oil. AB - The effect of modified atmosphere packaging (MAP) on the most important spoilage fungi of bread was investigated. Penicillium commune, P. roqueforti, Aspergillus flavus and Endomyces fibuliger were able to grow at oxygen levels down to 0.03%, while the chalk mould E. fibuliger was capable of growing even in the presence of an oxygen absorber. High levels of carbon dioxide retarded growth but not completely. As an alternative to MAP active packaging (AP) using volatile essential oils (EO) and oleoresins (OL) from spices and herbs were tested against a range of fungi commonly found on bread. Concentrations of 1, 10 or 100 microl EO or OL were added to a filter paper placed in the lid of a Petri dish inoculated with one of the test fungi. The Petri dish was sealed hermetically to avoid the exchange of gases. Mustard essential oil showed the strongest effect. Cinnamon, garlic and clove also had high activity, while oregano oleoresin only inhibited growth weakly. Vanilla showed no inhibitory effect towards the tested microorganisms at the applied concentrations. A. flavus was more resistant than the other microorganisms while P. roqueforti was the most sensitive. Mustard essential oil was investigated in greater detail. The minimal inhibitory concentration (MIC) for the active component, allyl isothiocyanate (AITC), was determined for the same species and an additional three moulds and one yeast. MIC values ranged from 1.8 to 3.5 microg/ml gas phase. Results showed that whether AITC was fungistatic or fungicidal depended on its concentration, and the concentration of spores. When the gas phase contained at least 3.5 microg/ml, AITC was fungicidal to all tested fungi. Results of sensory evaluation showed, that hot-dog bread was more sensitive to AITC than rye bread. The minimal recognisable concentration of AITC was 2.4 microg/ml gas phase for rye bread and between 1.8 and 3.5 microg/ml gas phase for hot-dog bread. These findings showed that the required shelf-life of rye bread could be achieved by active packaging with AITC. Active packaging of hot-dog bread, may nevertheless require the additional effect of other preserving factors to avoid off-flavour formation PMID- 11016612 TI - Properties of Bacillus cereus and other bacilli contaminating biomaterial-based industrial processes. AB - This paper is an overview on bacilli in industrial processes, with focus on food grade paper and paperboard production. Paperboards mainly contain sporeforming bacteria belonging to the genera Bacillus, Paenibacillus and Brevibacillus, usually found in quantities from < 50 to 250 cfu g(-1) homogenized paperboard. Of those frequently found, Bacillus cereus group, B. licheniformis, B. subtilis and Brevibacillus brevis are important for food hygiene because of their hydrolytic activities on food components and the ability of some strains to produce food poisoning toxins or to grow at refrigerated temperatures. We found that the phenotypic properties (lecithinase activity, nitrate reduction) used in standard methods (e.g., ISO, FDA, IDF) to recognize B. cereus, were unreliable for industrial isolates. Whole cell fatty acid composition of a group of the industrial isolates deviated so much from those in a widely used commercial database that the strains were not or only poorly recognized as B. cereus. Industrial isolates, including toxigenic ones, often missed one or more of these characters, even in cases where 100% 16S rDNA identity was found with B. cereus or with B. thuringiensis. 11-Methyldodecanoic acid and trans-9-hexadecenoic acid were found without exception in over 200 industrial B. cereus group isolates and in over 30 culture collection strains. The detection of these fatty acids is a secure method for the identification of B. cereus. Negative reaction for starch hydrolysis and for BCET-RPLA test and a specific ribotype were found in all B. cereus strains producing the emetic toxin. PMID- 11016613 TI - Development of bioactive food packaging materials using immobilised bacteriocins lacticin 3147 and nisaplin. AB - Immobilisation of the bacteriocins nisin and lacticin 3147 to packaging materials was investigated. Stability of both cellulose-based bioactive inserts and anti microbial polyethylene/polyamide pouches was examined over time. Anti-microbial activity against the indicator strain Lactococcus lactis subsp. lactis HP, in addition to Listeria innocua DPC 1770 and Staphylococcus aureus MMPR3 was observed for all bacteriocin-adsorbed materials. Activity retention of the inserts showed an initial decrease in the first week of storage but remained stable for the remaining 3 months of the trial. However, adsorption of lacticin 3147 to plastic film was unsuccessful, nisin bound well and the resulting film maintained its activity for 3-month period, both at room temperature and under refrigeration. When applied to food systems, the anti-microbial packaging reduced the population of lactic acid bacteria in sliced cheese and ham stored in modified atmosphere packaging (MAP) at refrigeration temperatures, thus extending the shelf life. Nisin-adsorbed bioactive inserts reduced levels of Listeria innocua by > or = 2 log units in both products, and Staphylococcus aureus by approximately 1.5 log units in cheese, and approximately 2.8 log units in ham. Similar reductions were observed in cheese vacuum-packaged in nisin-adsorbed pouches. PMID- 11016614 TI - Microbial diversity during maturation and natural processing of coffee cherries of Coffea arabica in Brazil. AB - The magnitude and diversity of the microbial population associated with dry (natural) processing of coffee (Coffea arabica) has been assessed during a 2-year period on 15 different farms in the Sul de Minas region of Brazil. Peptone water washed samples were taken of maturing cherries on trees (cherries, raisins and dried cherries) and from ground fermentations. The microbial load varied from 3 x 10(4) to 2.2 x 10(9) cfu/cherry with a median value of 1.6 x 10(7) cfu/cherry. The microbial load increased after heavy rainfall on cherries that were drying on the ground. At all stages, bacteria were usually the most abundant group, followed by filamentous fungi and finally yeasts. Counts of bacteria, yeasts and fungi varied considerably between farms and at different stages of maturation and processing and no consistent pattern could be seen. Yeasts showed an increase during the fermentation process. Median counts were not significantly different for fungi, yeasts and bacteria between the 2 years although Gram-negative bacteria dominated in the wet year and Gram-positive bacteria dominated in the dry year. Of a total of 754 isolates, 626 were identified to at least genus level comprising 44 genera and 64 different species. The 164 isolates of Gram-negative bacteria included 17 genera and 26 species, the most common of which were members of the genera Aeromonas, Pseudomonas, Enterobacter and Serratia. Of 191 isolates of Gram-positive bacteria, 23 were spore-forming and included six Bacillus species, and 118 were non-spore-formers of which over half were Cellulomonas with lesser numbers of Arthrobacter, Microbacterium, Brochothrix, Dermabacter and Lactobacillus. Of the 107 yeast isolates, 90 were identified into 12 genera and 24 different species and almost all were fermentative. The most common genera, in decreasing frequency, were Pichia, Candida, Arxula and Saccharomycopsis. There were many rarely described yeasts including Pichia lynferdii and Arxula adeninivorans. Almost all 292 fungal isolates were identified to genus level and 52 were identified to species level. Cladosporium, Fusarium and Penicillium each comprised about one third of the isolates and were found on all farms. Only 3% of the isolates were Aspergillus. Beauvaria, Monilia, Rhizoctonia and Arthrobotrys species were also occasionally found. The microbial flora is much more varied and complex than found in wet fermentations. The genera and species identified include members known to have all types of pectinase and cellulase activities. PMID- 11016615 TI - Analysis of the role of betL in contributing to the growth and survival of Listeria monocytogenes LO28. AB - Survival of the food-borne pathogen Listeria monocytogenes in environments of elevated osmolarity and reduced temperature is attributed, at least in part, to the accumulation of the trimethylammonium compound glycine betaine. Previously we identified betL, a gene encoding the secondary glycine betaine transporter BetL, which we linked to the salt tolerance of Listeria. In this report, we demonstrate that betL, preceded by a consensus sigmaB-dependent promoter, is regulated by osmotic up-shock, at least in part at the level of transcription. Using allelic exchange mutagenesis we constructed an in-frame deletion in betL, and used this mutant to determine the role of BetL in contributing to the growth and survival of L. monocytogenes, both in a high risk food (Camembert cheese) and animal model. Our results indicate that while BetL plays an important role in glycine betaine mediated osmoprotection, mutating the gene does not significantly effect either the cryotolerance or virulence of the organism. PMID- 11016616 TI - Stressed salmonella are exposed to reactive oxygen species from two independent sources during recovery in conventional culture media. AB - Previously, Stephens et al. [J. Appl. Microbiol. 83 (1997) 445-455] developed a sensitive technique for studying the resuscitation of low levels of stressed Salmonella. Using this technique the influence on recovery performance of the peptone component of buffered peptone water was investigated. Within 12 different peptone types as much as 3.5 log10 cells/ml difference was observed between the best and worst performing formulations. Poor recovery performance was linked to reactive oxygen species (ROS) generation through auto-oxidation of reducing sugars and photo-sensitisation of sensitive components such as riboflavin. Supplementary recovery agents were explored with only Oxyrase, which has both enzymes to degrade ROS and the ability to rapidly turn a medium anaerobic, having any significant effect. It improved the speed of recovery and increased, by up to 100-fold, the number of stressed cells recovered. Stressed cells were further studied by flow cytometry with cell sorting, based on the staining pattern from a novel fluorochrome combination, into good and poor recovery media. It was identified that within a stressed population the removal of all oxygen protected actively respiring cells the most by forcing them to generate energy from anaerobic metabolism thus avoiding any risk from accidental endogenous ROS generation. The recognition of two independent sources of oxidative stress in the routine use of conventional culture media is discussed in relation to pathogen detection and other areas of food microbiology. PMID- 11016617 TI - Estrogens, BRCA1, and breast cancer. AB - Findings obtained in in vitro assays and animal studies indicate that estrogens might influence the activity of the tumor suppressor gene BRCA1, and BRCA1 in turn may suppress the activity of the estrogen receptor. This review will discuss the possibility that interactions between estrogens and BRCA1 partly explain why elevated circulating estrogen levels appear to increase breast cancer risk among postmenopausal women but not among young women. A hypothesis is proposed that estrogens have a dual role in affecting breast cancer risk. In young women whose breasts have not yet accumulated critical mutations required for cancer initiation and promotion, activation of BRCA1 by estrogens helps to maintain genetic stability and induce differentiation, and therefore estrogens do not increase breast cancer risk. Breasts of older women, in contrast, are likely to contain transformed cells whose growth is stimulated by estrogens. Although BRCA1 is also probably activated by estrogens in older women, its function may have been impaired, for example, due to increased methylation associated with aging. Estrogen exposure in women who carry germ-line mutations in BRCA1 may always increase breast cancer risk because estrogens would be able to cause DNA damage and increase genetic instability without being opposed by BRCA1-induced repair activity. This might lead to an increase in the number of overall mutations, including those that initiate breast cancer. In addition to increasing genetic instability, reduced BRCA1 activity may also be linked to changes in the mammary gland morphology that predispose individuals to breast cancer. For example, a persistent presence of lobules type 1, which are the least differentiated lobular structures in the human breast, is seen in the BRCA1 mutation carriers. The aim of this review is to discuss the role of premenopausal estrogens in breast cancer and to initiate more research that would lead to novel means of reducing breast cancer risk, particularly among BRCA1 mutation carriers. PMID- 11016618 TI - p53 is essential for chemotherapy-induced hair loss. AB - Anticancer drugs stimulate apoptosis in the hair follicles (HF) and cause hair loss, the most common side effect of chemotherapy. In a mouse model for chemotherapy-induced hair loss, we demonstrate that p53 is essential for this process: in contrast to wild-type mice, p53-deficient mice show neither hair loss nor apoptosis in the HF keratinocytes that maintained active proliferation after cyclophosphamide treatment. HF in p53 mutants are characterized by down regulation of Fas and insulin-like growth factor-binding protein 3 and by increased expression of Bcl-2. These observations indicate that local pharmacological inhibition of p53 may be useful to prevent chemotherapy associated hair loss. PMID- 11016619 TI - Identification by cDNA microarray of genes involved in ovarian carcinogenesis. AB - To identify genes involved in the development or progression of ovarian cancer, we analyzed gene expression profiles of nine ovarian tumors using a DNA microarray consisting of 9121 genes. Comparison of expression patterns between carcinomas and the corresponding normal ovarian tissues enabled us to identify 55 genes that were commonly up-regulated and 48 genes that were down-regulated in the cancer specimens. When the five serous adenocarcinomas were analyzed separately from the four mucinous adenocarcinomas, we identified 115 genes that were expressed differently between the two types of tumor. Investigation of these genes should help to disclose the molecular mechanism(s) of ovarian carcinogenesis and define molecular separation of the two most common histological types of ovarian cancer. PMID- 11016620 TI - Micropthalmia transcription factor: a new prognostic marker in intermediate thickness cutaneous malignant melanoma. AB - Micropthalmia transcription factor (Mitf) is involved in melanocyte development and differentiation. The current study was undertaken to determine whether there is a relationship between Mitf expression and survival in patients with intermediate-thickness (1.0-4.0 mm) melanoma. Original paraffin blocks or slides of the primary tumor were accessible in 63 such patients. Mitf expression was evaluated by immunocytochemistry and analyzed visually. Slides were graded as follows according to the percentage of cells whose nuclei stained positive for Mitf: (a) 0, 0%; (b) +1, 1-25%; (c) +2, 26-50%; (d) +3, 51-75%; and (e) +4, > 75%. Median follow-up was 50 months. Mean thickness was 2.2 +/- 0.7 mm. Mean overall survival was 171.90 +/- 13.12 months. Mean disease-free survival was 168.53 +/- 13.96 months. Fifty-two melanomas (82.5%) stained positive for Mitf. By univariate analysis, mean overall survival and disease-free survival in patients whose melanomas did not express Mitf were 80.89 +/- 17.98 months (median, 51 months) and 71.36 +/- 19.87 months (median, 40 months), respectively. This compares with 187.90 +/- 13.41 months (median, not reached) and 186.78 +/- 13.84 months (median, not reached), respectively, for patients whose melanomas expressed Mitf (P = 0.0086 and P = 0.0054). These findings persisted in multivariate analysis. In addition, patients with > 50% Mitf expression had significantly fewer nodal metastases after node dissection than patients with < or = 50% Mitf expression (P = 0.04). Our data suggest that Mitf may be a new molecular prognostic marker in patients with intermediate-thickness melanoma. PMID- 11016621 TI - Population-based mapping of pulmonary adenoma susceptibility 1 locus. AB - Pulmonary adenoma susceptibility 1 (Pas1), the major locus affecting inherited predisposition to lung tumor development in mice, maps near the Kras2 gene. We previously reported a significant association between a KRAS2/RsaI polymorphism and the risk and prognosis of lung adenocarcinoma (ADCA) in the Italian population. In the present case-control study, we examined 269 lung ADCA patients, 121 squamous cell lung carcinoma patients, and 632 healthy individuals (general population controls) in the Japanese population with genetic markers spanning approximately 1200 kb in the KRAS2 region. Allele-specific oligonucleotide hybridization revealed the same KRAS2/RsaI polymorphism associated with risk and prognosis as in Italian lung ADCA patients; the polymorphism was significantly associated with clinical stage (P < 0.001) and survival rate (log rank = 0.0014), confirming the mapping of PAS1 and pointing to the role of this locus in human lung cancer. PMID- 11016622 TI - Fields of aberrant CpG island hypermethylation in Barrett's esophagus and associated adenocarcinoma. AB - Esophageal adenocarcinoma (EAC) is thought to develop through a multistage process in which Barrett's metaplasia progresses through low- and high-grade dysplasia to invasive cancer. Transcriptional silencing of tumor suppressor genes by promoter CpG island hypermethylation has been observed in many types of human cancer. Analysis of CpG island hypermethylation in EAC has thus far been limited to the CDKN2A (p16) gene. In this study, we extend the methylation analysis of EAC to include three other genes, APC, CDH1 (E-cadherin), and ESR1 (ER, estrogen receptor alpha), in addition to CDKN2A. Molecular analysis can provide insight into the complex relationships between tissues with different histologies in Barrett's esophagus and associated adenocarcinoma. Therefore, we have mapped the spatial distribution of methylation patterns in six esophagectomy cases in detail. Hypermethylation of the four CpG islands was analyzed by the MethyLight technique in 107 biopsies derived from these six patients for a total of 428 methylation analyses. Our results show that normal esophageal squamous epithelium is unmethylated at all four CpG islands. CDH1 is unmethylated in most other tissue types as well. Hypermethylation of ESR1 is seen at high frequency in inflammatory reflux esophagitis and at all subsequent stages, whereas APC and CDKN2A hypermethylation is found in Barrett's metaplasia, dysplasia, and EAC. When it occurs, hypermethylation of APC, CDKN2A, and ESR1 is usually found in a large contiguous field, suggesting either a concerted methylation change associated with metaplasia or a clonal expansion of cells with abnormal hypermethylation. PMID- 11016623 TI - Pleiotropic resistance to DNA-interactive drugs is associated with increased expression of genes involved in DNA replication, repair, and stress response. AB - A combination of four genetic suppressor elements (GSEs), two of which are derived from putative transcriptional regulators, was previously found to increase resistance to drugs inhibiting DNA replication in HT1080 fibrosarcoma cells. In the present study, two GSE-transduced cell lines, isolated with and without cytotoxic selection, were found to be resistant to a diverse group of DNA interactive agents, including aphidicolin, hydroxyurea, cytarabine, etoposide, doxorubicin, and mafosfamide. Changes in gene expression associated with GSE induced drug resistance were analyzed by cDNA array hybridization and reverse transcription-PCR. Twenty genes were found to be up-regulated in both of the resistant cell lines. These include genes involved in DNA replication and repair (e.g., PCNA, XRCC1, B-MYB, and GADD45), transcriptional regulators associated with stress response, and cell cycle checkpoint control (e.g., YB-1, DBPA, and ATF4), and genes for signal transduction proteins (e.g., protein tyrosine phosphatase 1B and regulatory subunits alpha and beta of cAMP-dependent protein kinase). The observed changes in gene expression may play a role in pleiotropic resistance to different classes of DNA-targeting drugs. PMID- 11016624 TI - The dual impact of coxsackie and adenovirus receptor expression on human prostate cancer gene therapy. AB - In a recent paper, we reported a significant difference in coxsackie and adenovirus receptor (CAR) from several human bladder cancer cell lines that correlated with their sensitivities to adenoviral infection (Y. Li, R-C. Pong, J. M. Bergelson, M. C. Hall, A. I. Sagalowsky, C-P. Tseng, Z. Wang, and J. T. Hsieh, Cancer Res., 59: 325-330, 1999). In human prostate cancer, CAR protein is down regulated in the highly tumorigenic PC3 cell line, which suggests that, in addition to its function as a viral receptor, CAR may have a pathophysiological role in prostate cancer progression. In this paper, we document that CAR does not merely enhance the viral sensitivity of prostate cancer cells but also acts as a tumor inhibitor for androgen-independent prostate cancer cells. Our data indicate that CAR is a potential therapeutic agent for increasing the efficacy of prostate cancer therapy. PMID- 11016625 TI - Ataxia telangiectasia-related protein is involved in the phosphorylation of BRCA1 following deoxyribonucleic acid damage. AB - The breast/ovarian cancer susceptibility gene BRCA1 exerts its tumor suppressor function, at least in part, by participating in DNA repair and/or DNA damage responsive pathways. BRCA1 protein is hyperphosphorylated following various DNA damaging events. Here, we report that the ataxia telangiectasia mutated protein related protein kinase (ATR) is involved in the phosphorylation of BRCA1 following gamma radiation and hydroxyurea treatment. We have shown that ATR can phosphorylate several BRCA1 fragments in vitro and that a kinase-inactive mutant of ATR interacts with BRCA1 in vivo. Taken together, these results suggest that ATR directly phosphorylates BRCA1 following DNA damage. PMID- 11016626 TI - The cyclooxygenase-2 inhibitor celecoxib is a potent preventive and therapeutic agent in the min mouse model of adenomatous polyposis. AB - Epidemiological and animal studies suggest that nonsteroidal anti-inflammatory drugs (NSAIDs) may reduce colon cancer risk. NSAIDs nonselectively inhibit both the constitutive cyclooxygenase (COX) 1 associated with side effects and the desired therapeutic target COX-2, which is induced in inflammation and neoplasia. We used the adenomatous polyposis coli (Apc) mutant Min mouse model to determine whether the selective COX-2 inhibitor celecoxib is effective for adenoma prevention and/or regression, and whether it might be safer than the nonselective NSAID previously shown to be most effective in this model, piroxicam. Min mice (n = 120) were randomized to treatment with celecoxib (0, 150, 500, or 1500 ppm celecoxib mixed in the diet) or piroxicam. To distinguish prevention from regression effects, groups were treated either "early" (before adenomas develop) or "late" (after most adenomas are established). Celecoxib caused dramatic reductions in both the multiplicity and size of tumors in a dose-dependent manner (P < 0.01). Early treatment with 1500 ppm of celecoxib was effective for prevention, decreasing tumor multiplicity to 29% and tumor size to only 17% of controls (P < 0.01). Late treatment demonstrated regression effects, reducing tumor multiplicity and size by about half. In contrast to the significant toxicity of piroxicam, which caused ulcers complicated by perforation and bleeding, celecoxib caused no gastrointestinal side effects and did not inhibit platelet thromboxane B2 at plasma drug levels similar to those obtained in early clinical trials in humans. These results provide the first evidence that selective inhibitors of COX-2 are safe and effective for the prevention and regression of adenomas in a mouse model of adenomatous polyposis and strongly support ongoing clinical trials in humans with the same syndrome. The broader population of patients with common sporadic adenomas that have somatic mutations of the same gene (APC) may also benefit from this treatment approach. PMID- 11016627 TI - Novel actions of the antitumor drugs vinflunine and vinorelbine on microtubules. AB - Vinflunine is a novel Vinca alkaloid presently in Phase I clinical trials. In preclinical studies, it exhibited superior antitumor activity to that of other Vinca alkaloids, including vinorelbine from which it was synthetically derived. Vinca alkaloids appear to inhibit cell proliferation by affecting the dynamics of spindle microtubules. Here we have analyzed the effects of vinflunine and vinorelbine on microtubule dynamic instability and treadmilling and found that these newer drugs exert effects on microtubule dynamics that differ significantly from those of the classic Vinca alkaloid, vinblastine. The major effects of vinflunine and vinorelbine on dynamic instability were a slowing of the microtubule growth rate, an increase in growth duration, and a reduction in shortening duration. In marked contrast to the action of vinblastine, they neither reduced the rate of shortening nor increased the percentage of time the microtubules spent in an attenuated state, neither growing nor shortening detectably. In addition, vinflunine and vinorelbine suppressed treadmilling, but less strongly than vinblastine. The diverse actions of these drugs on microtubules are likely to produce different effects on mitotic spindle function, leading to different effects on cell cycle progression and cell killing. Nontumor cells with normal checkpoint proteins may tolerate the relatively less powerful inhibitory effects of vinflunine and vinorelbine on microtubule dynamics better than the more powerful effects of vinblastine. Thus the unique constellation of effects of vinflunine and vinorelbine on dynamic instability and treadmilling may contribute to their superior antitumor efficacies. PMID- 11016628 TI - Cell-based screen for antimitotic agents and identification of analogues of rhizoxin, eleutherobin, and paclitaxel in natural extracts. AB - We describe a cell-based assay for antimitotic compounds that is suitable for drug discovery and for quantitative determination of antimitotic activity. In the assay, cells arrested in mitosis as a result of exposure to antimitotic agents in pure form or in crude natural extracts are detected by ELISA using the monoclonal antibody TG-3. The assay was used to screen >24,000 extracts of marine microorganisms and invertebrates and terrestrial plants and to guide the purification of active compounds from 5 of 119 positive extracts. A new rhizoxin analogue was found in a Pseudomonas species, six new eleutherobin analogues were identified from the octocoral Erythropodium caribaeorum, and two paclitaxel analogues were found in the stem bark of the tree Ilex macrophylla. The assay was also used for quantitative comparison of the antimitotic activity of different analogues. It revealed the importance of the C-11 to C-13 segment of the diterpene core of eleutherobin for its antimitotic activity. The identification of antimitotic compounds in very low abundance and their high (0.5%) occurrence in natural extracts indicates that drug discovery efforts using this cell-based assay may lead to the identification of structurally novel antimitotic agents. PMID- 11016629 TI - Inhibitory effect of citrus nobiletin on phorbol ester-induced skin inflammation, oxidative stress, and tumor promotion in mice. AB - The intake of citrus fruits has been suggested as a way to prevent the development of some types of human cancer. Nitric oxide (NO) is closely associated with the processes of epithelial carcinogenesis. We attempted a search for NO generation inhibitors in Citrus unshiu. The active constituent was traced by an activity-guiding separation. NO and superoxide (O2-) generation was induced by a combination of lipopolysaccharide and IFN-gamma in mouse macrophage RAW 264.7 cells, and by 12-O-tetradecanoylphorbol-13-acetate (TPA) in differentiated human promyelocyte HL-60, respectively. Expression of inducible NO synthase and cyclooxygenase 2 proteins were detected by Western blotting. The in vivo anti inflammatory and antitumor promoting activities were evaluated by topical TPA application to ICR mouse skin with measurement of edema formation, epidermal thickness, leukocyte infiltration, hydrogen peroxide production, and the rate of proliferating cell nuclear antigen-stained cells. As a result, nobiletin, a polymethoxyflavonoid, was identified as an inhibitor of both NO and O2- generation. Nobiletin significantly inhibited two distinct stages of skin inflammation induced by double TPA application [first stage priming (leukocyte infiltration) and second stage activation (oxidative insult by leukocytes)] by decreasing the inflammatory parameters. It also suppressed the expression of cyclooxygenase-2 and inducible NO synthase proteins and prostaglandin E2 release. Nobiletin inhibited dimethylbenz[a]anthracene (0.19 micromol)/TPA (1.6 nmol) induced skin tumor formation at doses of 160 and 320 nmol by reducing the number of tumors per mouse by 61.2% (P < 0.001) and 75.7% (P < 0.001), respectively. The present study suggests that nobiletin is a functionally novel and possible chemopreventive agent in inflammation-associated tumorigenesis. PMID- 11016630 TI - Differential activation of c-Jun NH2-terminal kinase and p38 mitogen-activated protein kinases by methyl methanesulfonate in the liver and brain of rats: implication for organ-specific carcinogenesis. AB - Methyl methanesulfonate (MMS), a direct-acting alkylating agent, is a strong brain carcinogen but a poor hepatocarcinogen in rats. To elucidate the mechanism(s) leading to tissue-specific carcinogenesis in response to MMS, we compared the activation of the stress-activated protein kinases (SAPKs), the c Jun NH2-terminal kinase (JNK) and p38, in the liver and brain of rats after i.p. injection of MMS. p38 was activated in both the liver and brain, but JNK was activated only in the liver in a dose- and time-dependent manner. The activation of JNK was preceded by the activation of SAPK or extracellular signal-regulated protein kinase kinase 1/mitogen-activated protein kinase kinase 4 in the liver, but no activation of SAPK or extracellular signal-regulated protein kinase kinase 1/mitogen-activated protein kinase kinase 4 was observed in the brain. The activation of JNK in the liver was accompanied by increased phosphorylation of activating transcription factor 2 and followed by an increase in the phosphorylation and level of c-Jun protein, in contrast to no such changes in the brain. To study the physiological consequences of these differential molecular events in the liver and brain, we examined MMS-induced apoptosis, a process shown to involve stress kinase activation. A significant increase in apoptotic cell death was detected in the liver but not in the brain after a MMS injection, which correlated with the patterns of JNK activation in the liver. Taken together, our results demonstrate that a tissue-specific signaling pathway(s) leading to distinct physiological responses in the liver and brain of rats exposed to MMS exists, suggesting a possible explanation for tissue-specific carcinogenic effects exerted by MMS in vivo. PMID- 11016631 TI - Human cytochrome P450 CYP2A13: predominant expression in the respiratory tract and its high efficiency metabolic activation of a tobacco-specific carcinogen, 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone. AB - The human CYP2A subfamily comprises three genes, CYP2A6, CYP2A7, and CYP2A13. CYP2A6 is active toward many carcinogens and is the major coumarin 7-hydroxylase and nicotine C-oxidase in the liver, whereas CYP2A7 is not functional. The function of CYP2A13 has not been characterized. In this study, a CYP2A13 cDNA was prepared by RNA-PCR from human nasal mucosa and was translated using a baculovirus expression system. In a reconstituted system, the expressed CYP2A13 was more active than CYP2A6 in the metabolic activation of hexamethylphosphoramide, N,N-dimethylaniline, 2'-methoxyacetophenone, and N nitrosomethylphenylamine but was much less active than CYP2A6 in coumarin 7 hydroxylation. Of particular interest, CYP2A13 was highly active in the metabolic activation of a major tobacco-specific carcinogen, 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone, with a catalytic efficiency much greater than that of other human cytochrome P450 isoforms examined previously. The tissue distribution of CYP2A13 was determined with isoform-specific RNA-PCR. CYP2A13 mRNA was detected in liver and a number of extrahepatic tissues, including nasal mucosa, lung, trachea, brain, mammary gland, prostate, testis, and uterus, but not in heart, kidney, bone marrow, colon, small intestine, spleen, stomach, thymus, or skeletal muscle. Quantitative PCR analysis further revealed that CYP2A13 mRNA is expressed at the highest level in the nasal mucosa, followed by the lung and the trachea. Together, these findings suggest that CYP2A13 plays important roles in xenobiotic toxicity and tobacco-related tumorigenesis in the human respiratory tract. PMID- 11016632 TI - Increased mutant frequency and altered mutation spectrum of the lacI transgene in Wilson disease rats with hepatitis. AB - The mutant strain Long-Evans Cinnamon (LEC) rat, which accumulates copper in the liver because of a mutation in the Atp7b gene, encoding a copper-ATPase, is a model of Wilson disease. It spontaneously develops hepatitis, and subsequently hepatocellular carcinoma and cholangiofibrosis. Excess intracellular copper has been thought to induce DNA damage through reactive oxygen species produced by Cu (II)/Cu (I) redox cycling, and also by direct interaction with DNA. We have developed lacI transgenic Wilson disease (WND-B) rats by mating LEC with Big Blue F344 rats carrying a lambda shuttle vector harboring the lacI gene. lacI mutations of the livers of C-B heterozygous (Atp7b w/m, lacI) and WND-B homozygous (Atp7b m/m, lacI) rats at 6, 24, and 40 weeks of ages were analyzed. Mutant frequencies in the WND-B rats were 2.0 +/- 0.7 x 10(-5), 5.3 +/- 0.9 x 10( 5), and 5.3 +/- 1.0 x 10(-5), respectively, significantly higher than those of C B rats. Nucleotide sequence analysis revealed that the frequency of deletion mutations of more than two nucleotides were much higher, 15% in WND-B rats, but only 2% in C-B rats. In addition, the average size of deletion was larger in the former. Loss of oligonucleotide-repeat units was specific and relatively frequent in WND-B rats. This type of mutation might be implicated in the induction of DNA strand scissions by reactive oxygen species. These findings suggest that the increase in mutant frequencies and/or the specific type of mutation according to copper accumulation play a crucial role in hepatocarcinogenesis in LEC rats. PMID- 11016633 TI - Lack of phenobarbital-mediated promotion of hepatocarcinogenesis in connexin32 null mice. AB - Connexin32 (Cx32) is the major gap junction forming protein in liver. We have recently shown that hepatocarcinogenesis is strongly enhanced in mice deficient in Cx32, demonstrating that lack of functional Cx32 accelerates liver tumorigenesis. Many tumor-promoting agents, including phenobarbital, block gap junctional intercellular communication in vitro, and it has been suggested that this effect is relevant for clonal expansion of neoplastic cells in vivo. We have now tested this hypothesis by analyzing the potency of phenobarbital as a liver tumor promoter in male Cx32-wild-type (Cx32(Y/+)) and Cx32-null (Cx32(Y/-)) mice. Preneoplastic and neoplastic liver lesions were induced in 6-week-old male mice by a single injection of 90 microg/g body weight of N-nitrosodiethylamine, and groups of mice were subsequently kept on phenobarbital-containing (0.05%) or control diet for 39 weeks. Frozen liver sections were prepared, and (pre)neoplastic lesions were identified by their deficiency in glucose-6 phosphatase staining. In addition, the number and size of macroscopically visible tumors were monitored. Phenobarbital led to a approximately 5-fold increase in the volume fraction occupied by glucose-6-phosphatase-deficient liver lesions in Cx32(Y/+) mice, whereas there was no such increase in Cx32(Y/-) mice. Even more pronounced differences were observed with respect to tumor response. Whereas phenobarbital clearly promoted the occurrence of numerous large hepatomas in Cx32(Y/+) mice, no such effect was seen in Cx32(Y/-) mice. These results demonstrate, for the first time, that functional Cx32 protein is required for tumor promotion by phenobarbital. PMID- 11016634 TI - Early resistance to therapy during induction in childhood acute lymphoblastic leukemia. AB - Many patients with acute lymphoblastic leukemia (ALL) are not cured by current therapy because of the development of drug resistance. It is not clear when resistance develops during the growth of the leukemic clone and whether resistant cells are already present at diagnosis or develop later during treatment. Twenty two uniformly treated children with ALL were studied throughout induction treatment. The size of the leukemic clone in blood and marrow was estimated by limiting dilution PCR analysis, using the rearranged immunoglobulin heavy chain gene as a molecular marker. The decline in the number of leukemic cells was biphasic in virtually all patients. For both marrow and blood, the logarithmic mean of the number of leukemic cells fell by approximately four orders of magnitude during the first 2 weeks, one order of magnitude during the third week, and not at all during the last two weeks of induction treatment. For marrow, the median of the fraction of leukemic cells in each patient that survived per week of treatment was 0.008 for the first 2 weeks, 0.12 for the third week, and 1.4 for the last 2 weeks; for blood, the corresponding figures were 0.003, 0.14, and 0.69, respectively. In individual patients, the results for marrow and blood showed good correlation. The biphasic decline of leukemic cell number suggests that most leukemic cells were sensitive to treatment and were rapidly killed, leaving behind a minor but substantial population of drug-resistant cells. The most likely explanation for this phenomenon is that these resistant cells were already present at diagnosis, their resistance having originated from genetic or epigenetic mutations during prior growth of the leukemic clone. PMID- 11016635 TI - Allosteric silencing of activating function 1 in the 4-hydroxytamoxifen estrogen receptor complex is induced by substituting glycine for aspartate at amino acid 351. AB - The active metabolite of tamoxifen, 4-hydroxytamoxifen (4-OHT), is used in the laboratory for mechanistic studies of antiestrogen action. This compound binds to the estrogen receptor alpha (ER) and silences activating function 2 (AF2) in the ligand binding domain, but activating function 1 (AF1) at the other end of the ER remains constitutive and is considered to be ligand independent. Amino acid D351 in the ligand binding domain appears to be critical for interactions with the antiestrogenic side chain of antiestrogens. We have devised an assay to evaluate the biological activity of 351 mutant ERs and antiestrogens at the transforming growth factor alpha (TGFalpha) gene in situ (J. I. MacGregor Schafer et al., Cancer Res., 59: 4308-4313, 1999). The substitution of glycine for aspartate at position 351 results in the conversion of the 4-OHT:ER complex from estrogen-like to completely antiestrogenic. In cells stably expressing D351G ER, the ER retains responsiveness to estradiol (E2) and also retains antiestrogenic responsiveness to both raloxifene and ICI 182,780. The relative binding affinity of E2 for D351G ER (0.77 +/- 0.17 x 10(-9) M) is comparable with wild-type ER (0.42 +/- 0.08 x 10(-9) M). In addition, the D351G ER retains the ability to bind SRC-1 in the presence of E2, thus D351G ER AF2 activity has not been compromised. We also used a cell line stably expressing an ER with a triple mutation in helix 12 (D538A, E542A, and D545A) that ablated AF2 activity, which resulted in decreased effects of E2, suggesting that both AF1 and AF2 activity are required for maximal estrogen activity in MDA-MB-231 cells. Interestingly, the triple mutation also completely reduced the estrogen-like actions of 4-OHT. We propose that a specific mutation at amino acid 351 can allosterically silence AF1 in the 4-OHT:ER complex by either preventing the binding of coactivators or encouraging the binding of a corepressor molecule. We suggest that the 4-OHT-specific site responsible for estrogen-like actions can be referred to as AF2b. This binding site would consist of at least four carboxylic acids at amino acids 351 and 538, 542 and 545 in helix 12 to permit coactivator docking for gene activation. The AF2b site is distinct from AF2 for E2 action. Further studies will provide insight into the estrogen-like actions of tamoxifen in select tissues and breast tumors and identify a significant mechanism of drug resistance to tamoxifen. PMID- 11016636 TI - Serum testosterone:estradiol ratio and the development of hepatocellular carcinoma among male cirrhotic patients. AB - The reason for the large male predominance in the occurrence of hepatocellular carcinoma (HCC) remains unknown, and sex hormones may contribute to this phenomenon. We examined possible associations of serum levels of testosterone, free testosterone, estradiol, sex hormone binding globulin, and testosterone:estradiol ratio (T:E2 ratio) with HCC development in a follow-up study of 46 Japanese male patients with liver cirrhosis predominantly of hepatitis C virus origin (76%). Serum samples were collected between December 1985 and December 1987, and the patients were completely followed until the end of 1995 for an average of 5.1 years. During the follow-up period, 20 patients (43%) developed HCC. Univariate analysis demonstrated that serum T:E2 ratio and testosterone were significant predictors of HCC; the hazard ratios (and 95% confidence intervals) in the middle and upper tertiles relative to the lower tertile were 2.0 (0.5-7.6) and 4.0 (1.1-14.6; P trend = 0.03) for T:E2 ratio and 0.8 (0.2-3.1) and 2.9 (1.0-8.5; P trend = 0.05) for testosterone. Adjustment for age, serum albumin, hepatitis virus markers, and other clinicobiological variables substantially increased the corresponding hazard ratios. In multivariate analysis, serum free testosterone appeared to be associated with increased risk, yet independent associations with estradiol and sex hormone binding globulin were not evident. These results indicate that elevated serum testosterone, together with decreased serum estrogens, may promote the development of HCC in cirrhosis. PMID- 11016637 TI - Polymorphic CAG and GGN repeat lengths in the androgen receptor gene and prostate cancer risk: a population-based case-control study in China. AB - The length of the polymorphic CAG trinucleotide repeat in the polyglutamine region of the androgen receptor (AR) gene is inversely correlated with the transactivation function of the AR. Because increased androgenic activity has been linked to prostate cancer and because an ethnic variation exists in the CAG repeat length, this polymorphism has been suggested to explain part of the substantial racial difference in prostate cancer risk. We conducted a population based case-control study in China to investigate whether CAG and other polymorphisms of the AR gene are associated with clinically significant prostate cancer in this low-risk population. Genomic DNA from 190 prostate cancer patients and 304 healthy controls was used for direct sequencing to evaluate the relationship of CAG and GGN (polyglycine) repeat length in the AR gene. Relative to western men, our study subjects had a longer CAG repeat length, with a median of 23 and only 10% of the subjects having a CAG repeat length shorter than 20. Men with a CAG repeat length shorter than 23 (median length) had a 65% increased risk of prostate cancer (odds ratio, 1.65; 95% confidence interval, 1.14-2.39), compared with men with a CAG repeat length of 23 or longer. For the GGN tract (GGT3GGG1GGT2GGCn), based on the sequencing results from 481 samples, we are the first to show that although GGC regions in the polyglycine tract are highly variable, there are no mutations or polymorphisms in the GGT and GGG regions. More than 72% of the subjects had a GGN repeat length of 23, and those with a GGN repeat length shorter than 23 had a 12% increased risk of prostate cancer (95% confidence interval, 0.71-1.78), compared with those with > or = 23 GGN repeats. Our study not only confirms that Chinese men do have a longer CAG repeat length than western men but also represents the first population-based study to show that even in a very low-risk population, a shorter CAG repeat length confers a higher risk of clinically significant prostate cancer. These results imply that CAG repeat length can potentially serve as a useful marker to identify a subset of individuals at higher risk of developing clinically significant prostate cancer. Larger studies are needed to evaluate the combined effect of CAG and GGN repeats. Because of the significance of AR in prostate cancer, investigation of factors that interact with the polyglutamine region of the AR gene to alter AR function and modulate prostate cancer risk is an important area for future research. PMID- 11016638 TI - Selective inhibition of vascular endothelial growth factor (VEGF) receptor 2 (KDR/Flk-1) activity by a monoclonal anti-VEGF antibody blocks tumor growth in mice. AB - Vascular endothelial growth factor (VEGF) is a multifunctional angiogenic growth factor that is a primary stimulant of the development and maintenance of a vascular network in embryogenesis and the vascularization of solid tumors. At the present time there are two well-characterized receptors for VEGF that are selectively expressed on endothelium. VEGF receptor 2 [VEGFR2 (KDR/Flk-1)] mediates endothelial cell mitogenesis and permeability increases, whereas the role of VEGF receptor 1 [VEGFR1 (Flt-1)] has not been clearly defined. In the present study, a monoclonal antibody, 2C3, is shown to block the interaction of VEGF with VEGFR2 but not with VEGFR1 through ELISA, receptor binding assays, and receptor activation assays. 2C3 blocks the VEGF-induced vascular permeability increase in guinea pig skin. 2C3 has potent antitumor activity, inhibiting the growth of newly injected and established human tumor xenografts in mice. These findings demonstrate the usefulness of 2C3 in dissecting the pathways that are activated by VEGF in cells that express both VEGFR1 and VEGFR2, as well as highlighting the dominant role of VEGFR2 in mediating VEGF-induced vascular permeability increase and tumor angiogenesis. PMID- 11016639 TI - Chemoprevention of prostate carcinogenesis by alpha-difluoromethylornithine in TRAMP mice. AB - Development of effective chemopreventive agents for human consumption requires conclusive evidence of their efficacy in animal models that have relevance to human diseases. Transgenic adenocarcinoma mouse prostate (TRAMP) is an excellent model of prostate cancer that mimics progressive forms of human disease inasmuch as 100% of males develop histological PIN by 8-12 weeks of age that progress to adenocarcinoma with distant site metastases by 24-28 weeks of age. In these animals, ornithine decarboxylase (ODC) activity (>3-fold) as well as protein expression (>4-fold) was found to be markedly higher in the dorsolateral prostate as compared with the nontransgenic littermates, suggesting their suitability to determine the chemopreventive effect of alpha-difluoromethylornithine (DFMO), an enzyme-activated irreversible inhibitor of ODC, against prostate cancer. Using male TRAMP mice, we studied the effect of oral consumption of DFMO on development of prostate carcinogenesis and surrogate end point biomarkers related to prostate cancer progression. In two independent experiments, each consisting of 8 animals on test, the cumulative incidence of prostatic cancer development at 28 weeks of age in 16 untreated TRAMP mice was 100% (16 of 16), whereas 94% (15 of 16) and 69% (11 of 16) of the animals exhibited distant site metastases to lymph nodes and lungs, respectively. Oral consumption of 1% DFMO (w/v) in the drinking water to TRAMP mice from 8 to 28 weeks of age resulted in a significant decrease in (a) weight (59%) and volume (66%) of prostate, (b) genitourinary weight (63%), and (c) ODC enzyme activity (52%) in the dorsolateral prostate. Importantly, in none of the DFMO-fed TRAMP mice were any distant metastases to lymph node and lungs observed. Furthermore, DFMO treatment resulted in the marked reduction in the protein expression of proliferation cell nuclear antigen, ODC, and probasin in the dorsolateral prostate. The protein expression of antimetastases markers, i.e., E-cadherin and alpha- and beta-catenin, was found to be restored in DFMO fed animals as compared with the non-DFMO-fed mice. These chemopreventive effects of DFMO were further confirmed by immunohistochemical analysis of the dorsolateral prostate. Histological analysis of the dorsolateral prostate of DFMO fed animals displayed marginal epithelial stratification, a small number of cribriform structures, elongated hyperchromatic epithelial nuclei, and a significant increase in apoptotic index. Non-DFMO-fed animals, on the other hand, displayed extensive epithelial stratification with profound cribriform structures accompanied with marked thickening, remodeling, and hypercellularity of the fibromuscular stroma. In nontransgenic littermates fed with DFMO, no significant alterations in the above parameters were evident. These data demonstrate that ODC represents a promising and rational target for chemoprevention of human prostate cancer and that TRAMP mice are excellent models for screening of novel drugs and chemopreventive regimens for potential human use. PMID- 11016640 TI - CD99 engagement: an effective therapeutic strategy for Ewing tumors. AB - CD99 is a Mr 32,000 transmembrane molecule that shows a high level of expression on cells of the hemopoietic system as well as on Ewing tumor cells. Within the hematopoietic system, CD99 has been implicated in cell adhesion and cell death, participating in this way in the differentiation of T-cell precursors. In this study, we demonstrate that engagement of CD99 significantly inhibits the in vitro and in vivo growth ability of Ewing tumor cells by delivering an apoptotic stimulus and reducing the malignant potential of these cells. Moreover, we show that anti-CD99 monoclonal antibodies may be advantageously used in association with conventional anticancer agents. These results provide a novel entry site for therapeutic intervention, which may have application in the care of patients with Ewing tumor, and warrant additional studies to clarify the molecular mechanisms activated by CD99 engagement. PMID- 11016641 TI - Intracranial inhibition of platelet-derived growth factor-mediated glioblastoma cell growth by an orally active kinase inhibitor of the 2-phenylaminopyrimidine class. AB - Glioblastoma multiforme is the most common primary human brain tumor, and it is, for all practical purposes, incurable in adult patients. The high mortality rates reflect the fact that glioblastomas are resistant to adjuvant therapies (radiation and chemicals), the mode of action of which is cytotoxic. We show here that an p.o.-active small molecule kinase inhibitor of the 2 phenylaminopyrimidine class may have therapeutic potential for glioblastomas. STI571 inhibits the growth of U343 and U87 human glioblastoma cells that have been injected into the brains of nude mice, but it does not inhibit intracranial growth of ras-transformed cells. Studies on a broad panel of genetically validated human and animal cell lines show that STI571 acts by disruption of the ligand:receptor autocrine loops for platelet-derived growth factor that are a pervasive feature of malignant astrocytoma. The cellular response of glioblastoma cells to STI571 does not appear to involve an apoptotic mechanism. PMID- 11016642 TI - Uroguanylin treatment suppresses polyp formation in the Apc(Min/+) mouse and induces apoptosis in human colon adenocarcinoma cells via cyclic GMP. AB - The enteric peptides, guanylin and uroguanylin, are local regulators of intestinal secretion by activation of receptor-guanylate cyclase (R-GC) signaling molecules that produce cyclic GMP (cGMP) and stimulate the cystic fibrosis transmembrane conductance regulator-dependent secretion of Cl- and HCO3-. Our experiments demonstrate that mRNA transcripts for guanylin and uroguanylin are markedly reduced in colon polyps and adenocarcinomas. In contrast, a specific uroguanylin-R-GC, R-GCC, is expressed in polyps and adenocarcinomas at levels comparable with normal colon mucosa. Activation of R-GCC by uroguanylin in vitro inhibits the proliferation of T84 colon cells and elicits profound apoptosis in human colon cancer cells, T84. Therefore, down-regulation of gene expression and loss of the peptides may interfere with renewal and/or removal of the epithelial cells resulting in the formation of polyps, which can progress to malignant cancers of the colon and rectum. Oral replacement therapy with human uroguanylin was used to evaluate its effects on the formation of intestinal polyps in the Min/+ mouse model for colorectal cancer. Uroguanylin significantly reduces the number of polyps found in the intestine of Min/+ mice by approximately 50% of control. Our findings suggest that uroguanylin and guanylin regulate the turnover of epithelial cells within the intestinal mucosa via activation of a cGMP signaling mechanism that elicits apoptosis of target enterocytes. The intestinal R-GC signaling molecules for guanylin regulatory peptides are promising targets for prevention and/or therapeutic treatment of intestinal polyps and cancers by oral administration of human uroguanylin. PMID- 11016643 TI - Human carbonyl reductase overexpression in the heart advances the development of doxorubicin-induced cardiotoxicity in transgenic mice. AB - Doxorubicinol (dxol) is the major metabolite formed in the hearts of cancer patients being treated with the widely used chemotherapeutic agent, doxorubicin (dox). The well-documented cardiomyopathy associated with dox treatment has been studied in vitro and ex vivo providing evidence that the C-13 hydroxy metabolite, dxol, might play a key role in the development of dox-induced cardiotoxicity. In this report, we have developed transgenic mice with heart-specific expression of human carbonyl reductase (HCBR), an enzyme that metabolizes dox to dxol. Dox was rapidly converted to dxol in the hearts of the transgenic expressers, which led to advanced development of both acute and chronic cardiotoxicity. Acute cardiotoxicity was evident by a 60% increase in serum creatine kinase activity and a 5-fold increase in cardiac damage measured by electron microscopy. Myofibril degeneration was the major damage observed in acute dox toxicity. Electrocardiograph telemetry, survival data, and electron microscopy were monitored during chronic dox-induced cardiotoxicity. HCBR expressers developed cardiotoxicity 6-7 weeks before the nonexpressers. The HCBR expressers survived for 5 weeks compared with 12 weeks for the controls. Electrocardiograph profiles and necropsies showed the cause of death to be the development of cardiomyopathies leading to congestive heart failure. Levels of dxol were four times higher in the HCBR expresser hearts than in the nonexpressers. Electron microscopy data showed swelling and major structural damage of the mitochondria in the HCBR expressers. These data demonstrate that the C-13 hydroxy metabolite of dox advances the development of dox-induced cardiotoxicity in an in vivo system and suggest that heart carbonyl reductase activity may contribute to dox induced cardiotoxicity in humans. PMID- 11016644 TI - Suberoylanilide hydroxamic acid, an inhibitor of histone deacetylase, suppresses the growth of prostate cancer cells in vitro and in vivo. AB - Suberoylanilide hydroxamic acid (SAHA) is the prototype of a family of hybrid polar compounds that induce growth arrest in transformed cells and show promise for the treatment of cancer. SAHA induces differentiation and/or apoptosis in certain transformed cells in culture and is a potent inhibitor of histone deacetylases. In this study, we examined the effects of SAHA on the growth of human prostate cancer cells in culture and on the growth of the CWR22 human prostate xenograft in nude mice. SAHA suppressed the growth of the LNCaP, PC-3, and TSU-Pr1 cell lines at micromolar concentrations (2.5-7.5 microM). SAHA induced dose-dependent cell death in the LNCaP cells. In mice with transplanted CWR222 human prostate tumors, SAHA (25, 50, and 100 mg/kg/day) caused significant suppression of tumor growth compared with mice receiving vehicle alone; treatment with 50 mg/kg/day resulted in a 97% reduction in the mean final tumor volume compared with controls. At this dose, there was no detectable toxicity as evaluated by weight gain and necropsy examination. Increased accumulation of acetylated core histones was detected in the CWR22 tumors within 6 h of SAHA administration. SAHA induced prostate-specific antigen mRNA expression in CWR22 prostate cancer cells, resulting in higher levels of serum prostate-specific antigen than predicted from tumor volume alone. The results suggest that hydroxamic acid-based hybrid polar compounds inhibit prostate cancer cell growth and may be useful, relatively nontoxic agents for the treatment of prostate carcinoma. PMID- 11016645 TI - Taxol mediates serine phosphorylation of the 66-kDa Shc isoform. AB - In the human lung carcinoma cell line A549, Taxol (20 nM) causes a decreased electrophoretic mobility of the 66-kDa Shc isoform (p66shc), beginning 4 h after drug exposure, and reaching a maximum at 9-18 h. No shift was observed for the 52 and 46-kDa isoforms of Shc. The electrophoretic mobility shift of p66shc caused by Taxol is not the result of tyrosine phosphorylation, and there is no indication of a Shc/Grb2 complex in Taxol-treated A549 cells. This modification is blocked by the serine/threonine protein phosphatase 2A. In vivo 32P-labeling and subsequent phosphoamino acid analysis of p66shc indicated that both the original and the shifted p66shc were predominantly serine phosphorylated. Cyanogen bromide digestion of p66shc produced a phosphorylated fragment with an apparent molecular weight of approximately 7.9 kDa from the untreated cells and two phosphorylated fragments, of approximately 7.9 and approximately 9.6 kDa, from the Taxol-treated cells. The domain of Taxol-induced serine phosphorylation is thought to be in the cyanogen bromide fragment containing residues 2-65. The Taxol-induced electrophoretic mobility shift of p66shc was inhibited by the protein synthesis inhibitor, cycloheximide, but not by the mitogen-activated and extracellular signal-regulated protein kinase kinase (MEK) inhibitor, PD98059. This mobility shift did not occur in Taxol-resistant A549-T12 cells treated with 20 nM Taxol. In addition to Taxol, other microtubule-interacting drugs caused a decreased electrophoretic mobility of p66shc. This Taxol-mediated serine phosphorylation seen in p66shc may result from a MEK-independent signaling pathway that is activated in cells that have a prolonged or abnormal mitotic phase of the cell cycle and may play a role in signaling events that lead to cell death. PMID- 11016646 TI - Noninvasive real-time monitoring of intracellular cancer cell metabolism and response to lonidamine treatment using diffusion weighted proton magnetic resonance spectroscopy. AB - We have used diffusion-weighted proton magnetic resonance spectroscopy (DWMRS) to noninvasively selectively observe only the intracellular metabolites of breast cancer and melanoma cell lines in vitro in real time. Breast cancer cell lines representing different stages in breast cancer progression were chosen for study. Intracellular biochemical profiles of six cell lines perfused in alginate beads were obtained. Spectral differences between groups of cell lines, including choline, lactate, and threonine peaks, were investigated. We also monitored response to the antineoplastic agent, lonidamine (LND), as a function of time and drug concentration in perfused cancer cells. Previous studies reported that this drug induced intracellular acidification and lactate accumulation. Diffusion weighted proton spectra demonstrated a 2- to 9-fold increase in the intracellular lactate signal as a response to LND treatment in several cancer cell lines. These results are consistent with the hypothesis that the principal mechanism of LND in some cancer cells is marked inhibition of lactate transport. Moreover, we have shown that there is a factor of two to three between the response of melanoma cells and that of some types of breast cancer cells. The higher sensitivity of the melanoma cells, as predicted by proton DWMRS, was correlated with changes in water-suppressed magnetic resonance spectra and confirmed by a biological assay. This study demonstrates the feasibility of using DWMRS for monitoring intracellular metabolism and for studying the effects and mechanisms of action of anticancer drugs. We believe that this method can be used for noninvasive clinical applications, such as the differentiation between benign and malignant tissue, real-time monitoring of response to therapy, dose response, and toxicity effects. PMID- 11016647 TI - Direct reversal of DNA damage by mutant methyltransferase protein protects mice against dose-intensified chemotherapy and leads to in vivo selection of hematopoietic stem cells. AB - Direct reversal of O6 adducts caused by chemotherapy agents is accomplished in mammalian cells by the protein O6-methylguanine DNA methyltransferase (MGMT). Some tumors overexpress MGMT and are resistant to alkylator therapy. One future approach to treatment of these tumors may rely on concurrent pharmacological depletion of tumor MGMT with O6-benzylguanine (6-BG) and protection of sensitive tissues, such as hematopoietic stem and progenitor cells, using genetic modification with 6-BG-resistant MGMT mutants. We have used retroviral-mediated gene transfer to transduce murine hematopoietic bone marrow cells with MGMT point mutants showing resistance to 6-BG depletion in vitro. These mutants include proline to alanine and proline to lysine substitutions at the 140 position (P140A and P140K, respectively), which show 40- and 1000-fold resistance to 6-BG compared with wild-type (WT) MGMT. Lethally irradiated mice were reconstituted with murine stem cells transduced with murine stem cell virus retrovirus expressing each mutant, WT MGMT, or mock-infected cells and then treated with a combination of 30 mg/kg 6-BG and 10 mg/kg 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) or with 40 mg/kg BCNU alone. Compared with mice treated with BCNU alone, significant myeloid toxicity and death occurred in mice reconstituted with mock infected or WT MGMT (<0.1 probability of survival) or the P140A mutant (0.13 probability of survival) MGMT cDNAs. In contrast, after an initial period of mild cytopenia, mice reconstituted with the P140K mutant (0.83 probability of survival) recovered nearly normal blood counts, even during continued treatment. Comparison of peripheral blood neutrophils after completion of 5 weekly treatments in these animals showed a direct correlation between the treatment and in vivo selection for progeny of transduced cells (pretreatment, approximately 8 12% transduced cells; no treatment, approximately 6% transduced cells; BCNU only, 51% transduced cells; 6-BG/BCNU, 93% transduced cells). To determine whether this selection occurred at the stem cell level, bone marrow from each treatment group was infused into secondary recipients. Whereas animals that received bone marrow from untreated animals reconstituted with 2% transduced cells, animals receiving marrow from 6-BG/BCNU-treated animals reconstituted with 94% transduced cells, demonstrating nearly complete selection for stem cells in the primary animals. Mice reconstituted with marrow from animals treated with BCNU only demonstrated 23% transduced cells, consistent with partial selection of stem cells in the primary mice. The levels of transduced cells also correlated with survival during a second round of intensive combination chemotherapy (probability of survival: 6 BG/BCNU, 1.0; BCNU alone, >0.70; no treatment, <0.1). These data demonstrate that mutant MGMT expressed in the bone marrow can protect mice from time- and dose intensive chemotherapy and that the combination of 6-BG and BCNU leads to uniform selection of transduced stem cells in vivo in mice. PMID- 11016648 TI - Role of Cyp1A1 in modulation of antitumor properties of the novel agent 2-(4 amino-3-methylphenyl)benzothiazole (DF 203, NSC 674495) in human breast cancer cells. AB - 2-(4-Amino-3-methylphenyl)benzothiazole (DF 203, NSC 674495) is a candidate antitumor agent with potent and selective activity against human-derived tumor cell lines in vitro and in vivo. Only sensitive cell lines (e.g., MCF-7) were able to accumulate and metabolize DF 203, forming the main inactive metabolite, 2 (4-amino-3-methylphenyl)-6-hydroxybenzothiazole (6-OH 203). Selective metabolism may therefore underlie its antitumor profile. DF 203 6-hydroxylase activity by MCF-7 cells was not constitutive but induced only after pretreatment of cells with DF 203, 3-methylcholanthrene, or beta-naphthoflavone. 6-Hydroxylation was strongly inhibited by either goat antirat cytochrome P450 1A1 (CYP1A1) serum or alpha-naphthoflavone. Both alpha-naphthoflavone and 6-OH 203 abrogated DF 203 induced growth inhibition. Microsomes from genetically engineered human B lymphoblastoid cells expressing CYP1A1, CYP1B1, or CYP2D6 metabolized DF 203 to 6 OH 203. Immunoblot analysis detected significantly enhanced CYP1A1 protein in a panel of sensitive breast cancer cell lines after exposure to DF 203. Neither constitutive expression nor induction of CYP1A1 protein was detected in nonresponsive breast (HBL 100, MDA-MB-435, and MCF-7/ADR) and prostate (PC 3 and DU 145) cancer cell lines. The expression of CYP1B1 was also modulated by DF 203 in the same sensitive cell lines. However, of the two isoforms, only CYP1A1 activity was irreversibly inhibited by DF 203 and significantly inhibited by 6-OH 203. In sensitive cell lines only, [14C]DF 203-derived radioactivity bound covalently to a Mr 50,000, protein which was immunoprecipitated by CYP1A1 antiserum. The covalent binding of [14C]DF 203 to recombinant CYP1A1 enzyme was NADPH-dependent and reduced by 6-OH 203 and glutathione. CYP1A1 appears essential for the metabolism of DF 203 and may have a pivotal, yet undefined, role in its antitumor activity. PMID- 11016649 TI - Pharmacological inhibition of phosphatidylcholine biosynthesis is associated with induction of phosphatidylinositol accumulation and cytolysis of neoplastic cell lines. AB - De novo production of phosphatidic acid (PA) in tumor cells is required for phospholipid biosynthesis and growth of tumor cells. In addition, PA production by phospholipase D has been cited among the effects of certain oncogenes and growth factors. In this report, it has been demonstrated that enhanced phospholipid metabolism through PA in tumor cells can be exploited pharmacologically for development of anticancer agents, such as CT-2584, a cancer chemotherapeutic drug candidate currently in Phase II clinical trials. By inhibiting CTP:choline-phosphate cytidylyltransferase (CT), CT-2584 caused de novo phospholipid biosynthesis via PA to be shunted away from phosphatidylcholine (PC) and into phosphatidylinositol (PI), the latter of which was doubled in a variety of CT-2584-treated tumor cell lines. In contrast, cytotoxic concentrations of cisplatin did not induce accumulation of PI, indicating that PI elevation by CT-2584 was not a general consequence of chemotherapy-induced cell death. Consistent with this mechanism of action, propranolol, an inhibitor of PA phosphohydrolase and phosphatidylcholine biosynthesis, was also cytotoxic to tumor cell lines, induced PI accumulation, and potentiated the activity of CT 2584 in cytotoxicity assays. As expected from biophysical properties of anionic phospholipids on cellular membranes, CT-2584 cytotoxicity was associated with disruption and swelling of endoplasmic reticulum and mitochondria. We conclude that CT-2584 effects a novel mechanism of cytotoxicity to cancer cells, involving a specific modulation of phospholipid metabolism. PMID- 11016650 TI - MT-21 is a synthetic apoptosis inducer that directly induces cytochrome c release from mitochondria. AB - We reported previously that a synthetic compound, MT-21, induced apoptosis by activating c-Jun-NH2-terminal kinase via the Krs/MST protein, which is activated by caspase-3 cleavage dependent on reactive oxygen species production. Here we examine the activation mechanism of caspase-3, an important cysteine aspartic protease, during MT-21-induced apoptosis. We found that MT-21 activated caspase-3 via caspase-9, but not via caspase-8. In addition, MT-21 induced the release of cytochrome c from the mitochondria that is necessary to activate caspase-9, and this release occurred before a change in membrane potential. This initiation process of MT-21-induced apoptosis was suppressed by overexpression of Bcl-2, which is known to prevent cells from undergoing apoptosis in response to a variety of stimuli. Moreover, when we treated mitochondria isolated from the cells with MT-21, the direct release of cytochrome c from the mitochondria was observed, whereas this effect was not observed in the mitochondria isolated from cells that overexpressed Bcl-2. Other apoptosis-inducing agents known to induce apoptosis via cytochrome c release from the mitochondria failed to release cytochrome c directly from isolated mitochondria. These findings indicate that MT 21 is a possible candidate antitumor agent that is able to induce apoptosis via the direct release of cytochrome c from the mitochondria. PMID- 11016651 TI - Use of two predictive algorithms of the world wide web for the identification of tumor-reactive T-cell epitopes. AB - Tumor cells can be effectively recognized and eliminated by CTLs. One approach for the development of CTL-based cancer immunotherapy for solid tumors requires the use of the appropriate immunogenic peptide epitopes that are derived from defined tumor-associated antigens. Because CTL peptide epitopes are restricted to specific MHC alleles, to design immune therapies for the general population it is necessary to identify epitopes for the most commonly found human MHC alleles. The identification of such epitopes has been based on MHC-peptide-binding assays that are costly and labor-intensive. We report here the use of two computer-based prediction algorithms, which are readily available in the public domain (Internet), to identify HL4-B7-restricted CTL epitopes for carcinoembryonic antigen (CEA). These algorithms identified three candidate peptides that we studied for their capacity to induce CTL responses in vitro using lymphocytes from HLA-B7+ normal blood donors. The results show that one of these peptides, CEA9(632) (IPQQHTQVL) was efficient in the induction of primary CTL responses when dendritic cells were used as antigen-presenting cells. These CTLs were efficient in killing tumor cells that express HLA-B7 and produce CEA. The identification of this HLA-B7-restricted CTL epitope will be useful for the design of ethnically unbiased, widely applicable immunotherapies for common solid epithelial tumors expressing CEA. Moreover, our strategy of identifying MHC class I-restricted CTL epitopes without the need of peptide/HLA-binding assays provides a convenient and cost-saving alternative approach to previous methods. PMID- 11016652 TI - Defining promiscuous MHC class II helper T-cell epitopes for the HER2/neu tumor antigen. AB - It is accepted that both helper and CTLs play a critical role in immune antitumor responses. Thus, the design of effective immune-based therapies for cancer relies in the identification of relevant tumor-associated antigens (TAAs) capable of eliciting strong helper and cytotoxic T-cell responses against tumor cells. The product of the HER2/neu oncogene is considered as a prototype TAA, because it is found overexpressed in a large variety of malignancies, whereas normal cells only produce low levels of this product. Several cytotoxic T-cell epitopes for HER2/neu have been identified that enable the design of peptide-based therapeutic vaccines for tumors expressing this TAA. Nevertheless, it is expected that inclusion of peptide epitopes capable of eliciting HER2/neu-specific T helper responses into these vaccines may enhance their effectiveness in the clinic. We describe here a strategy to identify helper T-cell epitopes for HER2/neu that focuses on peptides predicted to bind to numerous histocompatibility alleles (promiscuous epitopes), which would encourage their use in therapeutic vaccines for the general cancer patient population. Following this approach, we successfully identified several peptides that elicited T helper (CD4+) proliferative responses to peptides derived from HER2/neu. Most of the T-cell responses appeared to reflect a low affinity for antigen, which could be the result of immune tolerance because HER2/neu is expressed in low levels in normal cells and possibly including lymphocytes and monocytes. Interestingly, one of these peptides, HER2(883), was recognized by T cells in the context of either HLA DR1, HLA-DR4, HLA-DR52, and HLA-DR53, indicating a high degree of histocompatibility promiscuity. Furthermore, T cells that reacted with peptide HER2(883) could also recognize antigen-presenting cells that process HER2/neu recombinant protein. These results may be relevant for the design of more effective therapeutic vaccines for tumors expressing the HER2/neu oncogene product. PMID- 11016653 TI - In vitro characterization of radiolabeled monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen. AB - Prostate-specific membrane antigen (PSMA) is a well-characterized cell surface antigen expressed by virtually all prostate cancers (PCas). PSMA has been successfully targeted in vivo with (111)In-labeled 7E11 monoclonal antibody (mAb; ProstaScint; Cytogen, Princeton, NJ), which binds to an intracellular epitope of PSMA. This work reports the in vitro characterization of three recently developed mAbs that bind the extracellular domain of PSMA (PSMAext). Murine mAbs J415, J533, J591, and 7E11 were radiolabeled with 131I and evaluated in competitive and saturation binding studies with substrates derived from LNCaP cells. J415 and J591 were conjugated to 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid labeled with (111)In. The uptake and cellular processing of these antibodies were evaluated in viable LNCaP cells. All four mAbs could be labeled with 131I up to a specific activity of 350 MBq/mg with no or little apparent loss of immunoreactivity. Competition assays revealed that J415 and J591 compete for binding to PSMAext antigen. J533 bound to a region close to the J591 binding epitope, but J533 did not interfere with J415 binding to PSMA. mAb 7E11 did not inhibit the binding of J415, J533, or J591 (or vice versa), consistent with earlier work that these latter mAbs bind PSMAext whereas 7E11 binds the intracellular domain of PSMA. Saturation binding studies demonstrated that J415 and J591 bound with a similar affinity (Kds 1.76 and 1.83 nM), whereas J533 had a lower affinity (Kd, 18 nM). In parallel studies, all four mAbs bound to a similar number of PSMA sites expressed by permeabilized cells (1,000,000-1,300,000 sites/cell). In parallel studies performed with viable LNCaP cells, J415, J533, and J591 bound to a similar number of PSMA sites (i.e., 600,000-800,000 sites/cell), whereas 7E11 bound only to a subpopulation of the available PSMA sites (95,000 sites/cell). This apparent binding of 7E11 to viable cells can be accounted for by a 5-7% subpopulation of permeabilized cells produced when the cells were trypsinized and suspended. Up to five DOTA chelates could be bound to either J415 or J591 without compromising immunoreactivity. A comparison of the cellular uptake and metabolic processing of the 131I- and (111)In-labeled antibodies showed a rapid elimination of 131I from the cell and a high retention of (111)In. All four mAbs recognized and bound to similar numbers of PSMAs expressed by ruptured LNCaP cells (i.e., the exposed intracellular and extracellular domains of PSMA). By comparison to J415 and J591, J533 had a lower binding affinity. Both J415 and J591 recognized and bound to the same high number of PSMAs expressed by intact LNCaP. By contrast, 7E11 bound to fewer sites expressed by intact LNCaP cells (i.e., the exposed extracellular domain of PSMA). Both J415 and J591 are promising mAbs for the targeting of viable PSMA-expressing tissue with diagnostic and therapeutic metallic radionuclides. PMID- 11016654 TI - Loss of p27Kip1 from cyclin E/cyclin-dependent kinase (CDK) 2 but not from cyclin D1/CDK4 complexes in cells transformed by polyamine biosynthetic enzymes. AB - Cancer cells are known to display up-regulation of ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC), the key enzymes in the biosynthesis of polyamines that are essential for cellular proliferation. We have shown previously that overexpression of ODC or AdoMetDC alone can induce tumorigenic transformation of rodent fibroblasts. Because the subversion of normal cell cycle control is thought to be a crucial event in cancer development, we examined ODC- and AdoMetDC-transformed fibroblasts for alterations in the cell cycle components. The level of cyclin D1 and cyclin D1-dependent kinase and total cyclin-dependent kinase (CDK) 4 activities were elevated in the ODC transformants and particularly in the AdoMetDC transformants. Cyclin E content was not elevated, but a moderate increase in cyclin E-dependent kinase activity was seen in both cells. Total CDK2 activity was increased only in the ODC-transformed cells. The amount of the p27Kip1 CDK inhibitor was greatly decreased in both transformants. Nevertheless, p27Kip1 was present in the active cyclin D1/CDK4 complexes in the cells but absent from the cyclin E/CDK2 complexes. Restoration of p27Kip1 expression in the ODC- and AdoMetDC-transformed cells by transfection resulted in growth inhibition, but not in morphological reversion. An elevation in the level of hyperphosphorylated retinoblastoma protein was observed mainly in the ODC-transformed cells. These results suggest that the expression of ODC or AdoMetDC may affect cell cycle regulation in many ways. However, the largest common effect, which is therefore potentially relevant to some aspects of transformation, appears to be the constitutive down-regulation of p27Kip1 and its loss from the cyclin/CDK2 complexes. PMID- 11016655 TI - Tissue-specific carcinogenesis in transgenic mice expressing the RET proto oncogene with a multiple endocrine neoplasia type 2A mutation. AB - Germ line mutations of the RET proto-oncogene are responsible for the development of multiple endocrine neoplasia type 2A (MEN 2A), an inherited cancer syndrome characterized by medullary thyroid carcinoma, pheochromocytoma, and parathyroid hyperplasia. To study the mechanism of tissue-specific tumor development by RET with a MEN2A (cysteine 634-->arginine) mutation, we generated transgenic mice by introducing the RET-MEN2A gene fused to Moloney murine leukemia virus long terminal repeat. Expression of the transgene and its product was detected at variable levels in a variety of tissues including thyroid, heart, liver, colon, parotid gland, and brain. All of 29 mice analyzed developed thyroid C-cell hyperplasia or medullary carcinoma, accompanying high levels of serum calcitonin. In addition, development of mammary or parotid gland adenocarcinoma was observed in one-half of the transgenic mice. RET dimerization and its complex formation with Shc and Grb2 adaptor proteins were detected in tumor tissues. Unexpectedly, no tumor formation was found in other tissues despite RET-MEN2A expression where RET dimerization was undetectable. Because these tissues but not tumors expressed glial cell line-derived neurotrophic factor family receptor alpha (GFR alpha) at high levels, this suggested that GFR alpha expression may interfere in the dimerization of the RET-MEN2A mutant proteins, leading to tissue-specific tumor development in vivo. PMID- 11016656 TI - Involvement of cell surface glycans in adhesion of human colon carcinoma cells to liver tissue in a frozen section assay: role of endo-beta-galactosidase-sensitive structures. AB - Adhesion of human colon carcinoma variant cell lines expressing different levels of the cell surface sialyl Lewis X (sLeX) antigen to frozen sections of mouse liver was examined. KM12-HX cells that bound the monoclonal antibody (mAb) FH6 (anti-sLeX) and thus expressed a high level of sLeX demonstrated a greater degree of adhesion to liver sections than their low-binding counterparts, KM12-LX cells. The adhesion of KM12-HX cells to liver sections was partially blocked by mAb FH6, but not by another anti-sLeX mAb, KM93. The adhesion was Ca2+ dependent but was not inhibited by anti-E-selectin. Endo-beta-galactosidase treatment significantly reduced adhesion and resulted in the loss of cell surface binding sites for mAb FH6. O-linked oligosaccharides from KM12-HX cells incubated in the presence of p nitrophenyl-N-acetylgalactosaminide were fractionated by a combination of gel filtration, anion exchange chromatography, and normal phase high-performance liquid chromatography. The structure of a mAb FH6-reactive and endo-beta galactosidase-sensitive glycan was estimated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry in a post source decay mode and by glycosidase digestions to be NeuAc alpha2-3Gal beta1-4GlcNAc beta1-3Gal beta1 4Glc-NAc beta1-3Gal beta1-4(+/-Fuc alpha1-3)GlcNAc beta1-6(NeuAc alpha2-3Gal beta1-3)GalNAc-pNP. Mild detergent lysates of mouse liver surface-labeled with sulfo-NHS biotin were incubated with glutaraldehyde-fixed monolayers of KM12-HX cells, and bound components were isolated after EDTA treatment. A Mr 49,000 component that bound only to KM12-HX cells and not to KM12-LX cells was identified. PMID- 11016657 TI - Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. AB - Tumor cells may display a multidrug resistance phenotype by overexpression of ATP binding cassette transporter genes such as multidrug resistance (MDR) 1 P glycoprotein (P-gp) or the multidrug resistance protein 1 (MRP1). MDR3 P-gp is a close homologue of MDR1 P-gp, but its role in MDR is probably minor and remains to be established. The MRP1 protein belongs to a family of at least six members. Three of these, i.e., MRP1, MRP2, and MRP3, can transport MDR drugs and could be involved in MDR. The substrate specificity of the other family members remains to be defined. Specific monoclonal antibodies are required for wide-scale studies on the putative contribution of these closely related transporter proteins to MDR. In this report, we describe the extensive characterization of a panel of monoclonal antibodies (Mabs) detecting several MDR-related transporter proteins in both human and animal tissues. The panel consists of P3II-1 and P3II-26 for MDR3 P-gp; MRPr1, MRPm6, MRPm5, and MIB6 for MRP1; M2I-4, M2II-12, M2III-5 and M2III-6 for MRP2; M3II-9 and M3II-21 for MRP3; and M5I-1 and M5II-54 for MRP5. All Mabs in the panel appeared to be fully specific for their cognate antigens, both in Western blots and cytospin preparations, as revealed by lack of cross reactivity with any of the other family members. Indeed, all Mabs were very effective in detecting their respective antigens in cytospins of transfected cell lines, whereas in flow cytometric and immunohistochemical analyses, distinct differences in reactivity and suitability were noted. These Mabs should become valuable tools in studying the physiological functions of these transporter proteins, in screening procedures for the absence of these proteins in hereditary metabolic (liver) diseases, and in studying the possible contributions of these molecules to MDR in cancer patients. PMID- 11016658 TI - Increased expression of insulin-like growth factor I receptor in malignant cells expressing aberrant p53: functional impact. AB - We investigated the functional impact of p53 on insulin-like growth factor I receptor (IGF-IR) expression in malignant cells. Using the BL-41tsp53-2 cell line, a transfectant carrying temperature-sensitive (ts) p53 and endogenous mutant p53 (codon 248), we demonstrated a drastic down-regulation of plasma membrane-bound IGF-IRs on induction of wild-type p53. However, a similar response was obtained by treatment of BL-41tsp53-2 cells expressing mutant ts p53 with a p53 antisense oligonucleotide. Thus, even if the negative effect of wild-type p53 predominates under a competitive condition, these data indicate that mutant p53 may be important for up-regulation of IGF-IR. To further elucidate this issue, three melanoma cell lines (BE, SK-MEL-5, and SK-MEL-28) that overexpressed p53 were investigated. The BE cell line has a "hot spot" mutation (codon 248) and expresses only codon 248-mutant p53. SK-MEL-28 has a point mutation at codon 145. SK-MEL-5 cells did not exhibit any p53 mutations, but the absence of p21Waf1 expression suggested functionally aberrant p53. Our data suggest that interaction with Mdm-2 may underlie p53 inactivation in these cells. Using p53 antisense oligonucleotides, we demonstrated a substantial down-regulation of cell surface expression of IGF-IR proteins in all melanoma cell lines after 24 h. This was paralleled by decreased tyrosine phosphorylation of IGF-IR and growth arrest, and, subsequently, massive cell death was observed (this was also seen in BL 41tsp53-2 cells with mutant conformation of ts p53). Taken together, our results suggest that up-regulation of IGF-IR as a result of expression of aberrant p53 may be important for the growth and survival of malignant cells. PMID- 11016659 TI - Pleiotrophin can be rate-limiting for pancreatic cancer cell growth. AB - Pancreatic cancer is one of the most aggressive malignant tumors, with an overall survival rate of 2%. The identification of growth factors that contribute to the malignant phenotype can help to identify new targets for therapy. In this study, we analyzed the growth factor pleiotrophin (PTN) that was originally described as a developmentally regulated cytokine during early embryogenesis. More recently, PTN was found to be overexpressed in a variety of neuroectodermal tumors and described as an essential angiogenic growth factor in choriocarcinoma and melanoma, promoting metastatic growth. Recently, we discovered high expression levels of PTN in patients with gastrointestinal malignancies, particularly in those patients with pancreatic cancer. However, it is not known whether PTN is a contributor to the growth of pancreatic cancer or is only a bystander. We used ribozymes to deplete PTN mRNA from Colo357 pancreatic cancer cells and studied the resulting phenotype. The reduction of PTN resulted in a decrease in the proliferation rate, soft agar colony formation, and tumor growth in animals. Supplementation of cells with PTN partially reversed the ribozyme effect. The autocrine function of PTN was confirmed by using PTN-binding antibodies that inhibited the proliferation rate by 50% in Colo357 cells but also in a different pancreatic cancer cell line, Panc89. Our study identifies PTN as a new and essential growth factor for pancreatic cancer. Due to the restricted expression pattern of PTN in adults, PTN is suggested as a target for pancreatic cancer therapy. PMID- 11016660 TI - Mechanism for the antitumor and anticachectic effects of n-3 fatty acids. AB - Dietary intake of the n-6 fatty acid (FA) linoleic acid (LA) has a strong growth promoting effect on many rodent tumors and human tumor xenografts grown in immunodeficient rodents. n-3 FAs such as alpha-linolenic and eicosapentaenoic acids (EPAs), which differ from LA and arachidonic acid, respectively, by only a single double bond in the n-3 position, are recognized cancer chemopreventive and anticachectic agents. Understanding how this seemingly small structural difference leads to such remarkable functional differences has been a challenge. In a previous study, we showed that LA uptake, [3H]thymidine incorporation into DNA, and total DNA content were decreased in tissue-isolated hepatoma 7288CTC perfused in situ with arterial blood containing alpha-linolenic acid, EPA, or docosahexaenoic acids. The Ki for the inhibition of LA uptake and [3H]thymidine incorporation by alpha-linolenic acid was 0.18 and 0.25 mM, respectively. Here we show that the addition of alpha-linolenic acid or EPA to arterial blood inhibits tumor FA uptake, including LA, and the subsequent conversion of LA to the mitogen 13-hydroxyoctadecadienoic acid (13-HODE) in vivo and during perfusion in situ. [3H]Thymidine incorporation during perfusion in situ was also inhibited. Addition of 13-HODE to the arterial blood reversed the inhibition of [3H]thymidine incorporation but had no effect on FA uptake. These two n-3 FAs also inhibited FA transport in inguinal fat pads in vivo and during perfusion in situ in fed (FA uptake) and fasted (FA release) rats. The effects of EPA and talinolenic acid on transport of saturated, monounsaturated, and n-6 polyunsaturated FAs in hepatoma 7288CTC and inguinal fat pads during perfusion in situ were reversed by the addition of forskolin (1 microM), pertussis toxin (0.5 microg/ml), or 8-bromo cyclic AMP (10 microM) to the arterial blood. We conclude that the antitumor and anticachectic effects of n-3 FAs on hepatoma 7288CTC and inguinal fat pads in vivo result from an inhibition of FA transport. These inhibitions are mediated by a putative n-3 FA receptor via a Gi protein-coupled signal transduction pathway that decreases intracellular cyclic AMP. A specific decrease in LA uptake and its conversion to the mitogen 13-HODE causes the tumor growth inhibition. PMID- 11016661 TI - PCD1, a novel gene containing PDZ and LIM domains, is overexpressed in several human cancers. AB - In an effort to discover novel genes differentially expressed in human pancreatic cancer, we have identified a gene named PCD1 (pancreatic cancer derived) that is up-regulated in pancreatic dysplasia and cancer relative to normal pancreatic ductal epithelium. We cloned the full length (4572 bp) of this gene, which encodes a novel protein of 1064 amino acids containing a PDZ domain and a LIM domain. An alternatively spliced form with a deletion of 30 bp in the coding region was also found. In situ hybridization results showed that PCD1 is highly expressed in a significant percentage of colon, breast, liver, lung, pancreas, stomach, and prostate tumor tissues but is expressed in very few normal tissues. Northern blot hybridization confirmed the overexpression of PCD1 in colon and breast tumor tissues and also showed strong expression of PCD1 in the heart as well as in HeLa cells. Real-time quantitative reverse transcription-PCR verified the overexpression of PCD1 in primary colon tumors or in liver metastases relative to normal colon tissues in five of eight patients. The PCD1 gene maps to human chromosome 13q21.33. Because of its high levels of expression in neoplastic tissues and the presence of both PDZ and LIM domains, we suggest that PCD1 may play an important role in cytoskeletal reorganization during carcinogenesis. PMID- 11016662 TI - Prevention of phosphatidylinositol 3'-kinase-Akt survival signaling pathway during topotecan-induced apoptosis. AB - The serine/threonine kinase Akt (also known as protein kinase B) is a downstream effector of phosphatidylinositol-3'-kinase [PI(3)K] that is recognized as the major mediator of survival signals that protect cells from undergoing apoptosis. In the course of examining the target molecules of the topoisomerase I inhibitor topotecan, we found that topotecan treatment promoted Akt dephosphorylation that led to the inactivation of Akt in human lung cancer A549 cells. Transfection of the constitutively active akt cDNA into A549 cells resulted in the reduction of the cytotoxic effect of topotecan, indicating that inhibition of the Akt pathway played an important role in exhibition of topotecan-mediated cytotoxic effects. Further analysis of Akt dephosphorylation revealed that topotecan treatment suppressed upstream kinases of Akt, 3-phosphoinositide-dependent protein kinase 1, and PI(3)K. Overall, the results demonstrate that topotecan exhibited its cytotoxic effects by down-regulating the PI(3)K-Akt survival signaling pathway in addition to inhibiting topoisomerase I. PMID- 11016663 TI - Transactivation of the epidermal growth factor receptor in endothelin-1-induced mitogenic signaling in human ovarian carcinoma cells. AB - Endothelin (ET)-1 is produced in ovarian carcinoma cells and is known to act through ET(A) receptors as an autocrine growth factor in vitro and in vivo. In OVCA 433 human ovarian carcinoma cells, ET-1 caused phosphorylation of the epidermal growth factor receptor (EGF-R) that was accompanied by phosphorylation of Shc and its recruitment complexed with Grb2. These findings suggested that an EGF-R/ras-dependent pathway may contribute to the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (Erk) 2 and mitogenic signaling induced by ET-1 in these cells. Specific inhibition of EGF-R kinase activity by tyrphostin AG1478 prevented ET-1-induced transactivation of the EGF R, as well as Shc phosphorylation and recruitment with Grb2. Furthermore, ET-1 induced activation of Erk 2 was partially inhibited by tyrphostin AG1478. In accord with this finding, the mitogenic action of ET-1 in OVCA 433 cells was also significantly reduced by a concentration of tyrphostin AG1478 that abolished the growth response of EGF-stimulated cells. Inhibition of protein kinase C activity, which contributes to the proliferative action of ET-1 in OVCA 433 cells, had no effect on the activation of Erk 2 by ET-1, which suggests that this effect of protein kinase C does not involve ras-independent activation of Erk 2. Inhibition by wortmannin of PI3-kinase activity, which has been implicated in ET-1 and other G protein-coupled receptor (GPCR)-mediated signaling pathways, reduced Erk 2 activation by ET-1 but had no effect on ET-1-induced EGF-R and Shc phosphorylation. These findings indicate that ET-1-induced stimulation of Erk 2 phosphorylation, and mitogenic responses in OVCA 433 ovarian cancer cells are mediated in part by signaling pathways that are initiated by transactivation of the EGF-R. PMID- 11016664 TI - Quenching: fact or fiction? AB - Fragrance chemicals are a frequently reported cause of allergic contact dermatitis (ACD), a matter which has recently come into considerable prominence, to the point that legislation in Europe is under serious consideration. Certain skin-sensitizing fragrance chemicals have been reported by the producing industry to be rendered safe (quenched), at least in terms of ACD, when they are used in the presence of a specific quenching agent. Accordingly, it seemed timely to review this apparent quenching phenomenon, considering the available data and potential mechanistic hypotheses that might be used to explain it. If it is correct, it should be a phenomenon of potentially enormous value in the elimination of the allergenic properties of at least a proportion of common skin sensitizers. Whilst there is some evidence in man for the occurrence of quenching during the induction of skin sensitization, a much more substantial body of work has failed to find supportive evidence in various animals models, at a chemical level or at elicitation in human subjects with existing allergy. On balance, it is concluded that quenching of fragrance allergens is a phenomenon still awaiting positive evidence of existence. PMID- 11016665 TI - In-situ evaluation of barrier-cream performance on human skin using laser-induced breakdown spectroscopy. AB - Laser-induced breakdown spectroscopy (LIBS) was used to evaluate the effect of barrier creams (skin protective creams) on human skin. A Nd: YAG laser at 1,064 nm was used with a pulse energy of 100 mJ. A method was developed to measure the effectiveness of barrier creams against zinc ion absorption from aqueous zinc chloride solution and oil paste zinc oxide, which represent model hydrophilic and lipophilic metal compounds, respectively. Zinc was chosen since it posed no risk to human skin. 3 representative commercial barrier creams advertised as being effective against lipophilic and hydrophilic substances were evaluated by measuring zinc absorbed through the stratum corneum. 4 consecutive skin surface biopsies (SSB) were taken from biceps of the forearms of 6 volunteers at time periods of 0.5 h and 3 h after application of the protective cream. Results were compared with control skin where no barrier cream was used. The zinc atomic emission line at 213.9 nm was selected. Gate delay and gate width time was optimized to obtain the best signal-to-noise ratio (SNR) and precision. This method provided a facile and rapid screening of the effectiveness of skin barrier creams against zinc ion penetration. The barrier creams were shown to provide appreciable protection against the penetration of both ZnCl2 and ZnO into the skin. PMID- 11016666 TI - Benzalkonium chloride neutralizes the irritant effect of sodium dodecyl sulfate. AB - When benzalkonium chloride (BKC), a cationic surfactant, is added to sodium dodecyl sulfate (SDS), an anionic surfactant, and used in patch testing, on the basis of their known physicochemical interaction, it is possible to predict that there will be a tendency towards a reduction in the expected irritant response when compared to SDS alone. The aim of this study was to investigate whether BKC could reduce the irritant response to SDS when applied after the SDS exposure. 54 non-atopic adult volunteers were recruited for the study. 20% SDS was applied for 2 h under occlusion. 1% BKC was then applied to the same site. Various controls, including SDS application followed by water for 2 h, were included. The irritant reaction was assessed at 24 h and 48 h. 40 of the 54 subjects had some reaction when SDS was applied for 2 h followed by either benzalkonium chloride or water control under occlusion. In comparison to water control, where BKC was applied after SDS, 20 of the 40 responders had a weaker reaction but only 4 had a stronger response. This study shows that BKC applied to skin exposed to SDS attenuates the resulting irritant reaction. PMID- 11016667 TI - Immediate contact hypersensitivity induced by repeated hapten challenge in mice. AB - An immediate reaction was investigated during repeated challenge testing for contact hypersensitivity to dinitrofluorobenzene (DNFB) in BALB/c mice. The mice were sensitized to DNFB on back skin and repeatedly challenged with the same hapten on the left ear at 1 week intervals. The ear after the 5th challenge showed biphasic responses which consisted of an immediate and a delayed-type reaction. The reactions were hapten specific. Mast cell-deficient WBB6F1 W/WV mice did not show any immediate reaction, while congenic normal mice showed both immediate and delayed-type reactions. Histologically, numerous dermal mast cells were found in the left ear of repeatedly challenged BALB/c and WBB6F1 normal mice, while there were few mast cells in the ear of WBB6F1 W/WV mice. Anti-DNP IgE antibodies were detected in BALB/c, WBB6F1 normal and W/WV mice after repeated challenge with DNFB. Intradermal injection of anti-IgE antibodies in the repeatedly DNFB-challenged ear elicited an immediate reaction. These results suggest that immediate contact hypersensitivity develops through the production of anti-DNP IgE antibodies and an increase in dermal mast cells after repeated challenge with DNFB. PMID- 11016668 TI - Sensitizing potential of chlorothalonil in the guinea pig and the mouse. AB - The fungicide chlorothalonil is used extensively under several tradenames for the protection of various horticultural and fruit crops and bananas against fungal infections. It is also used as fungicide in wood preservation and as a preservative in paints. Clinical experience has shown chlorothalonil to be a contact allergen and several cases of allergic contact dermatitis attributed to chlorothalonil have been described. 2 previous guinea pig maximization test studies have shown the sensitizing potential of chlorothalonil to be high. The sensitizing property of chlorothalonil was studied by us with the predictive test methods the local lymph node assay and the cumulative contact enhancement test. In the local lymph node assay, chlorothalonil induced a dose-dependent increase in proliferation with a maximal stimulation index of 19.2 and 27.2. In the cumulative contact enhancement test, a statistically significant dose-dependent high sensitization rate was seen with a maximal sensitization rate of 100%. In conclusion, it is evident that chlorothalonil is an extremely potent contact allergen, inducing sensitization using only topical exposure on intact skin. PMID- 11016669 TI - Angry back syndrome is often due to marginal irritants: a study of 17 cases seen over 4 years. AB - 17 patients with angry back syndrome were identified over a 4-year period (1994 97). 10 such patients were classified as multiple reactors and 7 as exacerbations of atopic eczema. The 10 multiple reactors reacted most commonly to medicaments, rubber chemicals, fragrances, methylchloroisothiazolinone/methylisothiazolinone, colophonium and nickel. A final diagnosis of relevant allergic contact dermatitis was made in 8 of the 10 patients with multiple reactions. Relevant contact allergens were rubber chemicals, plants, metals and preservatives, all of which can be marginal irritants. None of the reactions in the patients with exacerbation of dermatitis were thought to be relevant or helpful in the further management of the patient. Reactions in the angry back syndrome appear to be due most commonly to marginal irritants. In multiple reactors, marginal irritants were also the most common allergens finally identified. PMID- 11016670 TI - Effect of test techniques of perfusion of neomycin sulfate patch tests. A comparative study with visual assessments. AB - Effects of test techniques on neomycin patch test results have not been thoroughly investigated. This study focuses on effects of dose and application time of neomycin sulfate patch tests on test results. The effects are assessed both visually and by perfusion. 16 subjects positive to neomycin are retested with neomycin sulfate in various doses and vehicles applied with different application devices for variable time intervals. All subjects were tested with a serial-dose series for time intervals ranging from 3 to 96 h (4 days [D]). The results show very good agreement between perfusion assessments and positive and negative visual assessments of reactions; however, a gray zone was detected where reactions with variable perfusion were associated with visually questionable test responses. 1:27 serial dilutions and wide application time intervals show a positive response with perfusion assessments and partly with visual assessments. Regardless of reading technique, highest reactivity of reactions was detected at either 96 h (day [D] 4) or 168 h (D7) in each subject with minor exceptions, regardless of dose, vehicle, application device or application time. Some findings of the study support the idea that reactions with papules alone are positive tests. Possible effect of tapes and foils on test results are discussed. PMID- 11016671 TI - Patch testing with neomycin sulfate. AB - The recommended patch test concentration of neomycin sulfate is 20% in petrolatum applied occluded for 48 h (2 days [D]). In the current study, the efficiency of such a test is compared to results with other techniques using lower allergen dose than obtained by approximately 20 microl of the 20% substance. Efficiency of petrolatum and demineralized water as vehicles are compared. 16 neomycin-sulfate positive subjects were retested with serial doses ranging from 0.4 to 0.0085 mg/cm2 neomycin sulfate in cellulose printed on polyester squares and applied by both tapes and plastic foils. Additionally, tests were performed with the 20% petrolatum substance in Finn Chambers. Tests containing the 20% substance in petrolatum and in water were applied directly with transparent foils. Readings ranged from 3 to 264 h (11D) following applications. Results show that foil applications of a polyester square dose, that is 7% of the dose obtained with the 20% substance, distinguished between perfusion of subjects who were visually positive and not with any test in the study. With the same test technique and visual assessments, 2 subjects were false-negative but developed doubtful test responses with the Finn Chambers. PMID- 11016672 TI - Allergic contact dermatitis from tertiary-butylhydroquinone (TBHQ) in a vegetable hydraulic oil. PMID- 11016673 TI - Contact allergy to individual fragrance mix constituents in relation to primary site of dermatitis. PMID- 11016674 TI - Persistent telangiectasia following patch testing with topical corticosteroids. PMID- 11016675 TI - Positive lesional patch tests in fixed drug eruptions from nimesulide. PMID- 11016676 TI - Contact dermatitis from 'ecological' textile dyes. PMID- 11016677 TI - Eyelid dermatitis from benzophenone used in nail enhancement. PMID- 11016678 TI - Protein contact dermatitis from pecan. PMID- 11016679 TI - Contact allergy due to propineb. PMID- 11016680 TI - Sensitizers commonly causing allergic contact dermatitis from cosmetics. PMID- 11016681 TI - Preliminary experience with a new "Mediterranean" Compositae mix. PMID- 11016682 TI - Photoallergic contact dermatitis from piketoprofen. PMID- 11016683 TI - The p53 molecule and its prognostic role in squamous cell carcinomas of the head and neck. AB - Despite intense research, the 5-year survival rate for patients with squamous cell carcinoma of the head and neck (SCCHN) is still low. Several different factors have been studied in the search for one or more factors that give important prognostic information at the time of diagnosis. Many recent studies have focused on the TP53 tumour suppressor gene, analysing its gene status and protein status. When looking at p53 protein expression, using immunohistochemistry, no correlation to patient outcome has been seen for the whole group of SCCHN. However, a significant association between p53 expression and poor patient outcome was found when looking only at patients with laryngeal squamous cell carcinomas. Also, in oral premalignant lesions, expression of p53 positive cells in the suprabasal layers of the epithelium has been seen as an indication of impending malignant development. Concerning the prognostic significance of mutations in the TP53 gene, results differ. But when restricting analysis to tumours with mutations causing an obvious change in protein, TP53 mutation was found to be a strong and independent variable for prognosticating survival. This review article gives an up-to-date overview of the p53 molecule and evaluates its possible prognostic role in SCCHN. Today it is clear that the p53 pathway is very important in SCCHN biology and potentially in its treatment. The function and importance of a few other cell cycle proteins connected to p53 are also discussed. PMID- 11016684 TI - Comparison of BrdU and cyclin A as markers of the S-phase in oral precancerous lesions. AB - A study comparing bromodeoxyuridine (BrdU) and cyclin A as markers of cells in the S-phase in oral precancerous lesions was performed. These were also compared with the growth fraction (GF) as assessed by Ki-67. Biopsies of 15 lesions were obtained, presenting clinically as leukoplakia or erythroplakia of the lateral tongue or floor of mouth. Half of each biopsy was incubated in BrdU and routinely fixed and processed. Sequential sections from each block were cut and stained immunohistochemically with antibodies against the following proteins: BrdU, Ki-67 and cyclin A. Stained sections were quantified and the labelling indices (LI) expressed per 100 of the total nucleated cell population (%) and per millimetre basement length (/mm). The mean LI% for BrdU was 11.24% (SD 2.83), for cyclin A it was 12.76% (SD 3.88) and the GF% was 29.25% (SD 11.88). The mean LI/mm for BrdU was 40.93/mm (SD 11.88), for cyclin A it was 47.59/mm (SD 18.28) and the GF/mm was 110.72/mm (SD 49.30). The BrdU and cyclin A indices were significantly correlated with each other. In the more dysplastic cases, the cyclin A LI was quantitatively much larger than that for BrdU, suggesting that the protein was being overexpressed. It was concluded that as a tool to study the kinetic aspects of the cell cycle in dysplastic lesions cyclin A was limited by the fact that it is overexpressed. In minimally dysplastic lesions and normal epithelia, however, cyclin A may be a viable alternative to BrdU for the study of the S-phase. PMID- 11016685 TI - First hints for a correlation between amplification of the Int-2 gene and infection with human papillomavirus in head and neck squamous cell carcinomas. AB - Infection with human papillomavirus (HPV) and alterations in certain genes have frequently been proposed as mechanisms in the pathogenesis of head and neck squamous cell carcinoma (HNSCC). Here, we investigated 47 HNSCC for the presence of HPV and, by fluorescence in situ hybridisation, for amplification of Int-2 and Hst-1 in the search for a possible correlation. The highest frequency of HPV infection was found in hypopharyngeal carcinomas, while amplification of Int-2 or Hst-1 was distributed more equally among the different localisations. Amplification of Int-2 was detectable (7 of 9 cases) in 78% of the HPV-positive carcinomas, whereas no virus infection could be found in the five cases with amplified Hst-1 only. In spite of the rather low number of infected tumour samples, our results suggest a correlation between HPV infection and amplification of Int-2. PMID- 11016686 TI - Oral and skin keratinocytes are stimulated to secrete monocyte chemoattractant protein-1 by tumour necrosis factor-alpha and interferon-gamma. AB - Monocyte chemoattractant protein-1 (MCP-1) is a member of the CC subfamily of chemokines. It is chemoattractant for monocytes, activated memory T cells and dendritic cells. We studied MCP-1 mRNA and protein production in cultured human oral keratinocytes (OK) and skin keratinocytes (SK). MCP-1 production was undetectable in resting keratinocytes. However, stimulation with tumour necrosis factors (TNF-alpha) or interferon-gamma (IFN-gamma) induced MCP-1 mRNA and protein synthesis in both cell types. Together, TNF-alpha and IFN-gamma acted synergistically to increase MCP-1 production. In each case MCP-1 production was greater for SK than OK. The kinetics of IFN-gamma-induced MCP-1 production were similar for both cell types, peaking around 72 h. In contrast, TNF-alpha induced a more rapid increase in MCP-1 production by SK than OK, peak production occurring after 24 and 72 h, respectively. IL-1alpha and IL-4 did not induce MCP 1 production. However, in combination with IFN-gamma, they induced a small extra increase in MCP-1 production in SK but not OK. PMID- 11016687 TI - Immunohistochemical study of the distribution of endogenous biotin and biotin binding enzymes in ductal structures of salivary gland tumours. AB - To clarify the pathologic value of endogenous biotin in the salivary gland, we examined in a series of neoplasms of the salivary gland by immunohistochemical staining the distribution of endogenous biotin and of biotin-binding enzymes, namely, acetyl CoA carboxylase (AC), which is a cytosolic enzyme, and pyruvate carboxylase (PC), which is a mitochondrial enzyme. In pleomorphic adenoma, we found biotin and PC in ductal epithelial elements, while AC was found mainly in myoepithelial elements. Carcinoma ex pleomorphic adenoma, adenocarcinoma and mucoepidermoid carcinoma were frequently immunopositive for biotin, PC and AC, while adenoid cystic carcinoma was rarely immunopositive for biotin, PC or AC. These results indicate that endogenous biotin might be associated with the mitochondrial enzyme, which is present at high levels in ductal cells of the salivary gland. However, the neoplastic cells in adenoid cystic carcinoma seemed to have an unusual expression of biotin and related enzymes. PMID- 11016688 TI - Sialochemical markers of salivary gland involvement with Sjogren's syndrome secondary to rheumatoid arthritis and primary biliary cirrhosis. AB - Sjogren's syndrome is an autoimmune condition affecting the lacrimal and salivary glands and can be associated with rheumatoid arthritis and primary biliary cirrhosis. Parotid salivas collected from patients and normal controls were analysed for lactoferrin, IgA and beta2-microglobulin (measured by ELISA), and cystatin (measured by a enzyme inhibition assay). Output data provided less variable means, whilst expressing results as a proportion of the total protein provided greater specificity as markers for Sjogren's syndrome. Levels of specificity for IgA, lactoferrin and beta2-microglobulin were all high (100, 95 and 100%, respectively). Sensitivity levels of these markers (but not cystatin) tended to be similar for Sjogren's syndrome secondary to primary biliary cirrhosis (IgA, 25%; lactoferrin, 63%; and beta2-microglobulin, 50%), compared to Sjogren's syndrome secondary to connective tissue diseases such as rheumatoid arthritis (IgA, 50%; lactoferrin, 86%; and beta2-microglobulin; 38%). PMID- 11016689 TI - Tooth eruption in a patient with craniometaphyseal dysplasia: case report. AB - Craniometaphyseal dysplasia (CMD) is a very rare genetic disorder of bone remodeling caused by osteoclast dysfunction. The clinical and radiographical features of oral findings are presented in a sporadic case of CMD in a child (age 10 years, 7 months). An intraoral examination showed severe malocclusions, including anterior crossbite and deep bite. Furthermore, a radiographic examination showed increased radiopacity of the maxilla and mandibular bones due to hyperostosis and sclerosis of the jaw. There was no root resorption of the canines or molars in the primary dentition, although root formation of the permanent teeth was proceeding. Dental age was calculated to be approximately 1 year, 4 months younger than his chronological age. The eruption speed of the permanent lateral incisors after the gingival emergence was shown to be within normal values, and we discuss whether the canines and premolars in the permanent dentition could erupt or not. PMID- 11016690 TI - Infection of clover by plant growth promoting Pseudomonas fluorescens strain 267 and Rhizobium leguminosarum bv. trifolii studied by mTn5-gusA. AB - Plant growth promoting Pseudomonas fluorescens strain 267, isolated from soil, produced pseudobactin A, 7-sulfonic acid derivatives of pseudobactin A and several B group vitamins. In coinoculation with Rhizobium leguminosarum bv. trifolii strain 24.1, strain 267 promoted clover growth and enhanced symbiotic nitrogen fixation under controlled conditions. To better understand the beneficial effect of P. fluorescens 267 on clover inoculated with rhizobia, the colonization of clover roots by mTn5-gusA marked bacteria was studied in single and mixed infections under controlled conditions. Histochemical assays combined with light and electron microscopy showed that P. fluorescens 267.4 (i) efficiently colonized clover root surface; (ii) was heterogeneously distributed along the roots without the preference to defined root zone; (iii) formed microcolonies on the surface of clover root epidermis; (iv) penetrated the first layer of the primary root cortex parenchyma and (v) colonized endophytically the inner root tissues of clover. PMID- 11016691 TI - Novel bacterial diversity recovered from the rhizosphere of oilseed rape (Brassica napus) determined by the analysis of 16S ribosomal DNA. AB - Soil was sampled to a distance of 2.5 mm beneath a root mat of oilseed rape (Brassica napus) in a model rhizosphere system. DNA was extracted and the 16S rDNA amplified, cloned and sequenced. Phylogenetic analysis of these sequences with those held on-line, revealed that 37% of the clones fell within the Holophaga /Acidobacterium phylum, 17% were within the proteobacteria, 14% of the clones were close relatives of Bacillus megaterium and 5% were related to Verrucomicrobium spinosum. An additional eleven clones (21%) could not be assigned to any known phylum and may represent novel bacterial lineages. This study highlights the diverse nature of rhizosphere soils and reinforces the role that molecular approaches play in unravelling such diversity. PMID- 11016692 TI - Roles of respiratory oxidases in protecting Escherichia coli K12 from oxidative stress. AB - Isogenic strains of Escherichia coli that were defective in either of the two major aerobic terminal respiratory oxidases (cytochromes bo' and bd) or in the putative third oxidase (cytochrome bd-II) were studied to elucidate role(s) for oxidases in protecting cells from oxidative stress in the form of H2O2 and paraquat. Exponential phase cultures of all three oxidase mutants exhibited a greater decline in cell viability when exposed to H2O2 stress compared to the isogenic parent wild-type strain. Cytochrome bo' mutants showed the greatest sensitivity to H2O2 under all conditions studied indicating that this oxidase was crucial for protection from H2O2 in E. coli. Cell killing of all oxidase mutants by H2O2 was by an uncharacterized mechanism (mode 2 killing) with cell growth rate affected. The expression of phi(katG-lacZ), an indicator of intracellular H2O2, was 2-fold higher in a cydAB::kan mutant compared to the wild-type strain at low H2O2 concentrations (< 100 microM) suggesting that cytochrome bd mutants were experiencing higher intracellular levels of H2O2. Protein fusions to the three oxidase genes demonstrated that expression of genes encoding cytochrome bd, but not cytochrome bo' or cytochrome bd-II was increased in the presence of external H2O2. This increase in expression of 4P(cydA-lacZ) by H2O2 was further enhanced in a cyo::kan mutant. The level of cytochrome bd determined spectrally and phi(cydA-lacZ) expression was 5-fold and 2-fold higher respectively in an rpoS mutant compared to isogenic wild-type cells suggesting that RpoS was a negative regulator of cytochrome bd. Whether the effect of RpoS is direct or indirect remains to be determined. PMID- 11016693 TI - Electrophoretic karyotypes of some related Mucor species. AB - Contour clamped homogeneous electric field (CHEF) gel electrophoresis was used to obtain electrophoretic karyotypes from nine Mucor strains representing five different species (M. bainieri, M. circinelloides, M. mucedo, M. plumbeus and M. racemosus). The chromosomal banding patterns revealed high variability among the isolates. The sizes of the DNA in the Mucor chromosomes were estimated to be between 2.5 and 8.7 Mb. The total genome sizes were calculated to be between 30.0 and 44.7 Mb. The applicability of these electrophoretic karyotypes for the investigation of genome structure, for strain identification and for species delimitation is considered. PMID- 11016694 TI - Mapping the distribution of 3-hydroxylipins in the Mucorales using immunofluorescence microscopy. AB - The distribution of endogenous 3-hydroxylipins (3-OH oxylipins) in representatives of the Mucorales was mapped using immunofluorescence microscopy. Strains of each of the following genera were examined: Absidia, Actinomucor, Cunninghamella, Mortierella (subgenus Micromucor), Mortierella (subgenus Mortierella), Mucor and Rhizomucor. Immunofluorescence microscopy was carried out using an antibody that was raised against 3R-hydroxy-5Z,8Z, 11 Z, 14Z eicosatetraenoic acid (3R-HETE), which cross-reacts with other 3-OH oxylipins. Subsequently, the occurrence and distribution of the antibody on the various reproductive stages of each fungus was noted. In Absidia, Actinomucor, Mortierella (subgenus Micromucor), Mucor and Rhizomucor, 3-OH oxylipins were found to be associated with the columellae and/or wall of the sporangium. In the representative of Cunninghamella, the 3-OH oxylipins were associated with the single-spored sporangiola. No 3-OH oxylipins were detected in the strains representing Mortierella (subgenus Mortierella). PMID- 11016695 TI - Hyperproduction of polyesters consisting of medium-chain-length hydroxyalkanoate monomers by strain Pseudomonas stutzeri 1317. AB - Pseudomonas stutzeri strain 1317 was found to grow on various fatty acids, alcohols, diols, as well as glucose and gluconate for the synthesis of polyhydroxyalkanoates (PHA) with various monomer units. The PHA monomer structures were dependent on the type of fatty acids and alcohols, as well as the diols in the culture media. Only even number monomers, such as 3-hydroxyhexanoate (HHx), 3-hydroxyoctanoate (HO) and 3-hydroxydecanoate (HD), were accumulated when even numbered fatty acids, alcohols, glucose and gluconate, as well as diol were used as carbon sources. Odd numbered fatty acids and odd numbered alcohols led to the formation of odd numbered monomers, such as 3-hydroxyvalerate (HV), 3 hydroxyheptanoate (HHp), 3-hydroxynonanoate (HN) and 3-hydroxyundecanoate (HU). The strain tolerated up to 1.5% of ethanol and made 8.3% of PHA when growth was conducted in 1.2% of ethanol. PHA formed up to 77% of cell dry weight when the strain was grown in tridecanoate. PHA synthesis was highly dependent on the nitrogen source. A depletion in nitrogen supply immediately resulted in PHA accumulation in cells grown in the glucose mineral medium. PMID- 11016696 TI - Renibacterium salmoninarum isolates from different sources possess two highly conserved copies of the rRNA operon . AB - The nucleotide sequences of the rRNA genes and the 5' flanking region were determined for R. salmoninarum ATCC 33209T from overlapping products generated by PCR amplification from the genomic DNA. Comparison of the sequences with rRNA genes from a variety of bacteria demonstrated the close relatedness between R. salmoninarum and the high G+C group of the actinobacteria, in particular, Arthrobacter species. A regulatory element within the 5' leader of the rRNA operon was identical to an element, CL2, described for mycobacteria. PCR, DNA sequence analysis, and DNA hybridisation were performed to examine variation between isolates from diverse sources which represented the four 16S-23S rRNA intergenic spacer sequevars previously described for R. salmoninarum. Two 23S-5S rRNA intergenic spacer sequevars of identical length were found. DNA hybridisation using probes complementary to 23S rDNA and 16S rDNA identified two rRNA operons which were identical or nearly identical amongst 40 isolates sourced from a variety of countries. PMID- 11016697 TI - Symbiosis of Astragalus cicer with its microsymbionts: partial nodC gene sequence, host plant specificity, and root nodule structure. AB - Astragalus cicer (cicer milkvetch) nodule bacteria were investigated for host plant specificity and partial nodC gene sequences, whilst their native host was studied for the microscopic structure of root nodules. The strains under investigation formed nodules not only on the original host but also on Astragalus glycyphyllos, Astragalus sinicus, Lotus corniculatus, and Phaseolus vulgaris. The nodules induced on the cicer milkvetch were classified as indeterminate and characterized by apical, persistent meristem, a large bacteroid region with infected and uninfected cells, and elongated bacteroids singly located inside peribacteroid membranes. By comparison of the partial nodC gene sequences of a representative strain of astragali rhizobia to those contained in the GenBank database, a close symbiotic relationship of A. cicer microsymbionts to Rhizobium sp. (Oxytropis) was found. PMID- 11016698 TI - Numerical phenetic study of the genus Carnobacterium. AB - Eighty-nine strains representing the genus Carnobacterium, Enterococcus durans, Vagacoccus salmoninarum and atypical Lactobacillus strains MT12 and MT13 were examined for 92 unit characters. Computer analysis of the data resulted in the recovery of four major, five minor and thirteen single membered clusters. Three cluster-groups contained seventy-four of the Carnobacterium strains, Enterococcus durans NCFB 596T and Lactobacillus maltaromicus NCFB 2382T. Cluster-group A was equated with Carnobacterium piscicola and cluster-group B with Carnobacterium divergens. Lactobacillus maltaromicus NCFB 2382T shared many properties in common with the C. piscicola strains. The recovery of several Carnobacterium strains as single membered clusters suggests that the genus Carnobacterium is underspeciated. Further work is also required to determine the subspecific structure of Carnobacterium divergens and Carnobacterium piscicola. PMID- 11016699 TI - RFLP analysis of the ribosomal internal transcribed spacers and the 5.8S rRNA gene region of the genus Saccharomyces: a fast method for species identification and the differentiation of flor yeasts. AB - The PCR amplification and subsequent restriction analysis of the region spanning the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene was applied to the identification of yeasts belonging to the genus Saccharomyces. This methodology has previously been used for the identification of some species of this genus, but in the present work, this application was extended to the identification of new accepted Saccharomyces species (S. kunashirensis, S. martiniae, S. rosinii, S. spencerorum, and S. transvaalensis), as well as to the differentiation of an interesting group of Saccharomyces cerevisiae strains, known as flor yeasts, which are responsible for ageing sherry wine. Among the species of the Saccharomyces sensu lato complex, the high diversity observed, either in the length of the amplified region (ranged between 700 and 875 bp) or in their restriction patterns allows the unequivocal identification of these species. With respect to the four sibling species of the Saccharomyces sensu stricto complex, only two of them, S. bayanus and S. pastorianus, cannot be differentiated according to their restriction patterns, which is in accordance with the hybrid origin (S. bayanus x S. cerevisiae) of S. pastorianus. The flor S. cerevisiae strains exhibited restriction patterns different from those typical of the species S. cerevisiae. These differences can easily be used to differentiate this interesting group of strains. We demonstrate that the specific patterns exhibited by flor yeasts are due to the presence of a 24-bp deletion located in the ITS 1 region and that this could have originated as a consequence of a slipped-strand mispairing during replication or be due to an unequal crossing-over. A subsequent restriction analysis of this region from more than 150 flor strains indicated that this deletion is fixed in flor yeast populations. PMID- 11016700 TI - Occurrence of Salmonella spp in estuarine and coastal waters of Portugal. AB - The presence of Salmonella and its relationship with indicator organisms of fecal pollution, such as total coliforms, fecal coliforms and fecal streptococci, was studied at two marine zones in Portugal. Seventeen different Salmonella serotypes were isolated and identified, S. virchow was the most frequently isolated (21.6%). In addition, a high percentage (35.1%) was recorded for some Salmonella serotypes of clinical significance, namely S. enteritidis, S. infantis, S. typhimurium and S. virchow. In any of the samples from the two zones Salmonella was not detected in the absence of any of the indicator organisms. However, the incidence of Salmonella as a function of indicator concentration intervals established by the EEC standards was 0, 10 and 19.3% at guide values of total coliforms, fecal coliforms and fecal streptococci, respectively in the Faro samples (south of Portugal). In contrast, Salmonella incidence rates of 37.5, 36.4 and 33.3% were recorded at the corresponding guide values the Caminha samples (north of Portugal). No significant correlations (p>0.005) were obtained between Salmonella and the indicators at the sampling stations; however, total coliforms and fecal streptococci were the indicators most closely related to Salmonella in Caminha and Faro samples, respectively. Survival experiments in Escherichia coli, Enterococcus faecalis and S. typhimurium, using diffusion chambers, were performed to verify whether the lack of correlation between indicators and Salmonella was due to different inactivation rates in seawater. The results indicate that survival percentages of the three microorganisms tested were similar after 48 h of exposure to seawater. PMID- 11016701 TI - State of the art of sentinel node biopsy in oncology. AB - Sentinel node biopsy has become a standard diagnostic procedure to assess lymph node status of various tumors. The combination of blue dye and a radioactive tracer offers the best chances of identifying the sentinel lymph node. Most progress in the technique of the sentinel node procedure has been made in melanoma and breast cancer. In melanoma, sentinel node biopsy has been introduced as a fundamental procedure for staging. Information on the lymphatic drainage from a melanoma can have a direct impact on the surgery. More recently, the technique has been successfully introduced in the management of breast cancer, in which a large number of unnecessary axillary dissections could be avoided. However, there are many other potential fields of application of the sentinel node biopsy (e.g. endometrial, vulvar, head and neck cancers) that are worthy of investigation. In any case, multicenter trials are required to standardize the procedures, taking into account several variables such as particle size and mode of delivery of the radiotracer, amount of radioactivity administered, number and location of injections, and choice of the hand-held probe. We briefly describe the technical and historical aspects of the sentinel node biopsy and summarize the main clinical trials proposed and/or performed in the field. PMID- 11016702 TI - Utility fo gallium-67 in determining the radicality of lymph node dissections of the neck for metastatic lymph nodes with extranodal spread in patients affected by carcinoma of the head and neck. AB - AIMS AND BACKGROUND: Based on the fact that scintigraphy more readily reveals uptake of a radioisotope in a superficial position owing to incomplete surgical radicality, the authors examined by 67Ga scintigraphy a group of patients who had undergone dissection of lymph nodes of the neck from carcinoma with extranodal spread. They then checked the follow-up to ascertain the efficacy of 67Ga scintigraphy in relation to the eventual recurrences in the soft tissues of the neck. METHODS: A group of 136 patients were examined by 67Ga scintigraphy and followed for a minimum of 3 years after complementary radiotherapy. A group of 20 patients with no lymph node metastases was used as control to evaluate eventual false positives or false negatives. RESULTS: Recurrences in the soft tissues of the neck occurred in 35 (42.7%) of the 82 patients positive at 67Ga scintigraphy and in 6 (11.1%) of the 54 patients negative at the examination (P = 0.0001). All the patients in the control group were negative at 67Ga scintigraphy and without recurrences. CONCLUSIONS: 67Ga scintigraphy can give reliable information on the risk of recurrences in the soft tissues of the neck. Since in spite of postoperative radiotherapy the percentage of local recurrences in cases with positive 67Ga scintigraphy was rather high (42.7%), the authors propose a scheme of radiotherapy based on administration of a higher dose per fraction on the scintigraphically positive area. PMID- 11016703 TI - Surgical therapy of T1 and selected cases of T2 glottic carcinoma: cordectomy, horizontal glottectomy and CO2 laser endoscopic resection. AB - AIMS AND BACKGROUND: Among the different laryngeal neoplasms, glottic carcinoma is known to be one of the most suitable for functional management. Nevertheless, the best treatment for T1 and T2 glottic carcinoma, whether an open neck procedure, endoscopy or radiotherapy, with reference to recurrence, survival, and functional results, has long been debated. STUDY DESIGN: From February 1983 to September 1997, 83 patients with well to undifferentiated glottic carcinoma (48 pT1a, 14 pT1b, and 21 selected cases of pT2 with impairment of vocal cord mobility) were submitted to surgery at the Otorhinolaryngologic Section of the Department of Surgical Sciences and Organ Transplantations of Cagliari University. Surgical treatment included 30 laryngofissures with simple or enlarged cordectomy, 22 horizontal glottectomies and 31 endoscopic laser resections. A retrospective review of the records of the patients was performed in order to obtain a better understanding of the outcome of the three different surgical procedures in our institution. RESULTS: According to the Kaplan-Meier method, the probability of remaining free of local recurrence 3 years after primary surgery was 0.90 for the T1 group and 0.85 for the T2 group. The distribution of recurrences for cordectomy, glottectomy and CO2 laser at 3 years showed a cumulative probability of remaining free of disease after primary surgery of 0.86, 0.85 and 0.88. The probability of remaining free of local recurrence 3 years after salvage surgery was 0.96 for the T1 group and 0.95 for the T2 group. Analyzing the phenomena for type of surgical procedure, local control at 3 years after salvage surgery for cordectomy, glottectomy and exclusive CO2 laser was 0.93, 0.90 and 0.92, respectively. In the endoscopic group, local control rate after any type of salvage therapy modified the percentage at 3 years to 100%. Anterior commissure spread (AC1-AC2) resulted in a difference (not statistically significant) in local control between the group of patients without and with anterior commissure involvement. Laryngeal preservation was achieved in 93.7% (45/48) of patients who survived after salvage surgery following open neck procedures and in 100% of patients originally submitted to the endoscopic approach. CONCLUSIONS: In our experience, although open laryngeal procedures can be still considered a valid option in the treatment of T1 and selected cases of T2 glottic carcinoma, endoscopic laser excision offered an oncologically adequate alternative to the traditional techniques, with minimum discomfort for the patient and satisfactory preliminary functional results. PMID- 11016704 TI - Cisplatin plus vinorelbine as a salvage regimen in refractory breast cancer. AB - AIMS AND BACKGROUND: Breast cancer refractory to known effective agents is one of the major clinical problems frequently encountered in practice. Cisplatin and vinorelbine are known to be active drugs in anthracycline-refractory cases. In this phase II study, the effectiveness and tolerability of cisplatin and vinorelbine was investigated when used in combination as a salvage regimen in the treatment of metastatic refractory breast cancer. STUDY DESIGN: Twenty-four patients with advanced refractory breast cancer who had been previously treated with a regimen containing doxorubicin were included in the study. Six of the 24 patients also received taxanes after failure of doxorubicin. Cisplatin at 80 mg/m2 on day 1 and vinorelbine at 25 mg/m2 on days 1 and 8 were given every 3 weeks. RESULTS: A total of 98 cycles of chemotherapy was given, with a median of 4/patient. The response rate was 25% (2 [8.3%] complete and 4 [16.7%] partial responses). The median survival rates were 14 months in responders and 5.5 months in nonresponders (P = 0.0282). One complete and one partial response were observed in patients previously treated with paclitaxel (overall response rate, 33%). The median response duration was 12.5 mo (range, 4-21) in complete and 4.5 mo (range, 1.5-13) in the partial response group. Grade 3 and 4 neutropenia occurred in 9 patients, with no toxic deaths. Grade 2-3 nausea and vomiting in 6 patients and grade 1 neuropathy in 1 patient were noted. CONCLUSIONS: Although the number of cases is insufficient to indicate that the combination will be effective, it is noteworthy in consideration of anthracycline and taxane refractory cases. A combination of cisplatin and vinorelbine seems to be a reasonable and acceptable choice as an alternative salvage regimen in such cases. PMID- 11016705 TI - Low-dose induction radiotherapy for stomach conservation in patients with massive gastric lymphoma. AB - Low-dose induction radiotherapy was performed in 4 patients with massive gastric lymphoma in order to treat the disease without gastrectomy. Following the radiotherapy, gastric lesions had shrunk considerably without any complications. Standard chemotherapy and/or radiotherapy could then be performed safely, and stomach conservation could be achieved in all 4 patients. PMID- 11016706 TI - Two case reports of vinorelbine extravasation: management and review of the literature. AB - Vinorelbine tartrate (Navelbine, Burroughs Wellcome Company, Research Triangle Park, NC, USA) is a semisynthetic vinca alkaloid approved in many countries for the first-line treatment of patients with advanced non-small-cell lung cancer. It is also used in the treatment of advanced breast and ovarian cancers and lymphoma. Like other vinca alkaloids, Navelbine can cause skin necrosis as a consequence of inadvertent extravasation in surrounding tissues during intravenous administration. In such cases, early treatment is strongly advocated. There is no documented case of vinorelbine extravasation in the literature. The authors herein report two cases successfully treated with hyaluronidase injections plus saline flushout under local anesthesia. PMID- 11016707 TI - Localization of the sentinel lymph node in breast cancer by combined lymphoscintigraphy, blue dye and intraoperative gamma probe. AB - Axillary lymph node status represents the most important prognostic factor in patients with operable breast cancer. A severe limitation of this technique is the relatively high rate of false negative sentinel lymph nodes (>5%). We studied 284 patients suffering from breast cancer; 264 had T1 tumors (16 T1a, 37 T1b and 211 T1c), while 20 had T2 tumors. All patients underwent lymphoscintigraphy 18-h before surgery. At surgery, 0.5 mL of patent blue violet was injected subdermally, and the sentinel lymph node (SN) was searched by gamma probe and by the dye method. The surgically isolated SN was processed for intraoperative and delayed examinations. The SN was successfully identified by the combined radioisotopic procedure and patent blue dye technique in 278/284 cases (97.9%). Analysis of the predictive value of the SN in relation to the status of the axillary lymph nodes was limited to 191 patients undergoing standard axillary dissection irrespective of the SN status. Overall, 63/191 (33%) identified SNs were metastatic, the SN alone being involved in 37/63 (58.7%) patients; a positive axillary status with negative SN was found in 10/73 (13.7%) patients with metastatic involvement. In T1a-T1b patients the SN turned out to be metastatic in 9/53 patients (17.0%). In 7/9 patients the SN was the only site of metastasis, while in 2/9 patients other axillary lymph nodes were found to be metastatic in addition to the SN. None of the 44 patients in whom the SN proved to be non-metastatic showed any metastatic involvement of other axillary lymph nodes. Our results demonstrate a good predictive value of SN biopsy in patients with breast cancer; the predictive value was excellent in those subjects with nodules smaller than 1 cm. PMID- 11016709 TI - Sentinel lymph node identification in breast cancer: feasibility study. AB - The aim of the study was to evaluate in our institute the technique of sentinel node (SN) identification and biopsy in the surgical treatment of early breast cancer. Between June 1998 and November 1999 54 patients (age range, 31-75 years) where studied. Inclusion criteria were age less than 75 years, indication for conservative surgery, absence of palpable axillary nodes, Karnofksy index >70. Lymphoscintigraphy was performed 16-18 hours prior to surgery, following injection of 0.1-0.2 mL of 99mTc-Nanocoll: the administered activity was 3-4 MBq in group A (44 pts) and 7-8 MBq in group B (10 pts). The colloids were administered by transdermal supralesional injection in 49 patients with palpable nodules and by intraparenchymal ultrasound-guided injection in five patients with non-palpable nodules. Planar projections were performed starting from the 5th until the 80th min (or 180th in the event of late migration). In 10 patients further projections were acquired 14-18 h following tracer administration. All nodes identified by gamma probe (MR 100 Pol.Hi.Tech) were histologically evaluated by immunohistochemistry and standard histology. Scintigraphic visualization of the SN was obtained in 49 patients: in 38 of these patients there was only one SN while in 11 patients there were two or three SNs. The delayed scan made in 10 patients did not show any further nodes. In all patients given US-guided perilesional injections migration was late (after at least 60 min). Our study confirms the validity of the scintigraphic procedure, its safety for patients and health care workers, and the feasibility of interdisciplinary collaboration. PMID- 11016708 TI - Sentinel lymph node and breast cancer staging: final results of the Turin Multicenter Study. AB - AIM OF THE STUDY: Validation of the sentinel node (SN) technique in breast cancer by means of lymphoscintigraphy. MATERIALS AND METHODS: From December 1996 to January 1999 102 T1-T2 breast carcinoma cases were recruited in Turin. 99mTc human serum albumin colloids were injected subdermally the day before surgery (mean activity, 5.2 +/- 2.5 MBq). Scintigraphic imaging was performed after injection. After identification of the SN during surgery by a hand-held gamma probe, the SN was excised and sent for histologic examination. SN histology was compared with that of other axillary nodes. RESULTS: The SN detection rate was 86.3%; among 88 cases with an identified SN, 37 (42%) had axillary metastases; the SN was metastatic in 35 cases (sensitivity, 94.6%); in 51.3% of pN+ cases (19/37) the SN was the only metastatic site. In two of the 53 negative SNs, SN histology did not match with that of the remaining axilla (negative predictive value, 96.2%; staging accuracy, 97.7%). CONCLUSIONS: Our results agree with those reported in the literature; however, except in clinical trials and experienced structures axillary lymph node dissection should not be abandoned when mandatory for prognostic purposes, considering that at present SN biopsy alone is not completely accurate for axillary staging, especially in the absence of an adequate learning period. PMID- 11016710 TI - Different sites and modes of tracer injection for mapping the sentinel lymph node in patients with breast cancer. AB - Several studies have been published describing the techniques of identification of the "sentinel lymph node" (SN). There are marked differences in the techniques used by different investigators. Although agreement exists about the tracer particle size and the volume of injection, it is unknown what is the best site where to inject the tracer or the vital dye. The aim of the present study was to define the influence of different sites of injection on imaging of the lymphatic ducts and their SNs. We performed a pilot study in 30 consecutive patients with breast cancer who underwent SN biopsy by means of radioguided surgery and vital blue dye mapping. The patients were divided into six groups of five patients each; each patient was given a subdermal (ID) or peritumoral (IP) injection of radiotracer (300 microCi in 150 mL of 99mTc-HSA microcolloids; Albures, Amersham Sorin) above the tumor site in order to localize the SN. After the identification of the SN, a second injection of radiotracer was performed, which was different in each patient subset. In some cases more than one lymph node appeared on the lymphoscintigraphic scans after the second injection of radiotracer. When the peritumoral route was used it took longer to visualize the lymphatic pathways. For the ID route, injection at the exact skin projection over the tumor is optimal. Internal mammary lymph nodes were identified by both IP (2) and ID (1) injection, irrespective of the quadrant in which the tracer was injected. Our findings support the hypothesis of a precise topographic correspondence between the primary tumor and its specific SN. The subdermal route is more accurate than the intraparenchymal route, as it allows faster identification of the lymphatic vessels and SN. We believe these observations should be taken into account for the proper selection of the injection site of either vital dye or radiopharmaceuticals. PMID- 11016711 TI - Sentinel node biopsy in breast cancer: the experience of Brescia Civic Hospital. AB - The accuracy of the sentinel node technique in the evaluation of axillary node involvement in breast cancer was evaluated in 83 consecutive patients with monofocal T1-2 carcinoma, who were clinically N0 and who underwent lymphoscintigraphy with 99mTc-colloid integrated with intraoperative sentinel node detection by a portable probe. Lymphoscintigraphy revealed at least one sentinel node in 75 patients (90.4%), always identified by the probe. In eight patients (9.6%) the sentinel node was detected neither by lymphoscintigraphy nor by the probe. All removed lymph nodes were analyzed by hematoxylin-eosin histology and the sentinel node by immunostaining. In 28/75 patients (37.3%) at least one metastatic axillary lymph node was detected; in 16 of the 28 N+ subjects (57%) only the sentinel node was positive. The false negative rate (sentinel node negative/other axillary lymph nodes positive) was 17.85% (5/28 patients). In 9/23 patients (39%) micrometastases were found in the sentinel node only. In conclusion, specific sentinel node positivity in 57% of cases supports the validity of the sentinel node concept. Moreover, nine patients would have been considered N0 by standard hematoxylin-eosin histology without sentinel node aided immunostaining. A 17.8% false negative rate calls for caution in patients with negative sentinel nodes. PMID- 11016712 TI - The success rate of identification of the sentinel lymph node in breast cancer: our training series. AB - A new phase of breast conserving surgery has started very recently, aimed at eliminating axillary dissection in node-negative patients by using the sentinel lymph node (SN) technique. Between November 1998 and January 2000 we performed 151 operations for breast cancer on 145 patients. We performed axillary lymphoscintigraphy using 99Tc-labeled human serum albumin microcolloidal particles injected subdermally in 50 patients who met our selection criteria. In this series we focused on the success rate of scintigraphic and surgical sentinel node identification. The number of scintigraphic identifications of the SN was 44 (88%). Only forty-three cases were evaluable, as in one case mapping showed an internal mammary hot node. All SNs were located at the first level. After removal of the SN complete axillary dissection was performed. Eighteen patients (41.8%) had metastatic disease in the axilla. There were five (11.6%) false negatives: two in T2 tumors, one in a T4 tumor and two in T1c tumors. We consider this series as our training series. Our results are similar to those reported in the literature. We believe that the most reasonable approach to SN biopsy is a two step procedure: the ideal candidates are patients with T1 cancer who can undergo the operation in an outpatient setting under local anesthesia and sedation. Complete axillary dissection is performed only if paraffin sections and immunohistochemistry show metastatic disease. PMID- 11016713 TI - Sentinel node study in early breast cancer. AB - Since October 1997 60 patients with early breast cancer (T <3 cm) were studied. All patients underwent lymphoscintigraphy with two types of colloid: the first (17 pts) with a particle size <1,000 nm; the second (43 pts) with a particle size <80 nm. The standard procedure consists of injection, on the day before surgery, of 70 MBq of the smaller nanocolloid in 0.4 cc saline divided over four sites, around the lesion or subdermally around the surgical scar. We utilize a low energy, high-resolution LFOV camera for scintigraphy and a probe specific for the sentinel node during surgery. In 56/60 patients (93.3%) lymphoscintigraphy showed the sentinel node (SN). In two cases the SN was not detected presumably because of lymphatic interruption by an old surgical scar; in the other two cases the sites of injection were too close to the SN, thus masking it. In five cases (9%) the SN was not visualized with the surgical probe but in two of these drainage to the internal mammary chain was observed. The apparently lower sensitivity of intraoperative localization was due to the extra-axillary lymphatic drainage or to the vicinity of the SN to the primary lesion. The SN proved to be metastatic in 12 cases. No false-negative SNs were found. In five cases (10%) the radiolabeled lymph node was the only node containing tumor cells (micrometastases): this result depends on the combined use of hematoxylin-eosin and rapid cytokeratin staining. The application of blue dye was useful for easier identification of the SN but did not allow detection of more SNs. Our preliminary results are extremely encouraging. Considering that at the early stages of breast cancer the likelihood of lymph node metastases is low (20% in our series) and no false negative were reported in this study, we conclude that with SN biopsy axillary lymph node dissection can be avoided, making surgery less aggressive but maintaining accuracy. PMID- 11016714 TI - Study of sentinel lymph node in breast cancer: our experience in 46 cases. AB - At the Surgery Department of Biella 46 patients were enrolled in a study on the sentinel lymph node (SN) in the period from 1 January 1999 to 30 September 1999. The aim of the study was to determine, on the basis of our own experience, the percentages of accuracy and concordance, and compare them with case series abroad and in Italy; in addition, we sought to establish a possible correlation between certain features of breast cancer and positivity of the axilla. The method utilized was lymphoscintigraphy and gamma probe. Fifteen cases with positive axillary lymph nodes and 12 cases with positive sentinel lymph nodes were found; there were no false positive and three false negative results. No migration of the tracer was observed with lymphoscintigraphy in two cases. The percentage of concordance obtained was 93.2% in the complete series and 96.5% in the subseries that excluded the learning curve. Comparing the percentage of concordance of our case series with those abroad and in Italy, an average overlapping percentage was obtained. The percentage of accuracy obtained in our study was 95.7%, which is slightly higher than the average of percentages of the case series abroad and in Italy. PMID- 11016715 TI - A study of sentinel node biopsy in T1 breast cancer treatment: experience of 48 cases. AB - Axillary clearance in breast cancer has been proven to be unnecessary in more than 50% of cases. Sentinel node biopsy (SNB) is a new technique that can be used to avoid unnecessary axillary clearance in breast cancer surgery. Our integrated team, consisting of surgeons, pathologists and nuclear medicine physicians, studied 48 cases of T1 breast cancer with lymphoscintigraphy-guided SNB. Before starting this study, the team performed 20 SNBs as a learning procedure. 500 microCi of 99mTc-nanocoll in 0.2 mL were injected around the lesion, under US or x-ray guidance if necessary. Static images in anterior, lateral and lateral oblique view collected at the end of a 20 min dynamic study were used to mark the SN on the skin. During surgery a gamma probe was used to guide SN resection, and node invasion was assessed with cytokeratin immunohistochemistry. In 14 patients tracer uptake was observed in a single node, in 30 patients in 2-4 nodes, whereas in four patients the nodes were scintigraphically missed. Surgical resection was possible in 42 nodes out of 54. All but two patients with negative immunohistochemistry for cancer cell clusters showed metastasis-free axillary nodes. All patients with positive SNBs (13) showed involved axilla. In four patients the lymphatic drainage was towards the intramammary chain; one node was juxtaclavicular and one node was intramammary in the upper outer quadrant. The overall sensitivity of the method was about 80%, the specificity about 90% with a diagnostic accuracy about 80%. SNB is a promising method for surgical decision making regarding axillary clearance in breast cancer. Adequate training of an interdisciplinary team is needed in order to successfully perform SNB and assess SN invasion. Its unusual anatomic location can be encountered and technical care is necessary to correctly identify and remove them. PMID- 11016716 TI - Lymphoscintigraphy and gamma probe tracing in detecting breast cancer lymph node involvement: can they replace axillary lymph node dissection? AB - BACKGROUND: Axillary lymph node status is the most important pathological determinant of prognosis in early breast cancer. However, axillary lymph node dissection (ALND) performed for pathological assessment is not without costs and morbidity. Recently, radioisotope-guided sentinel node biopsy (SNB) has been proposed as a promising technique for staging breast cancer patients. AIM OF THE STUDY: In this study we report our experience (76 patients) in radioguided sentinel node (SN) biopsy in breast cancer. The study was divided into two phases: the first represents our learning curve, necessary to establish our guidelines for its use in clinical practice, while the second phase was aimed at assessing the feasibility of SN localization using preoperative lymphoscintigraphy and intraoperative gamma probe (GP) detection. METHODS: All patients underwent lymphoscintigraphy (LS) up to two hours after tracer delivery (99mTc-micro-nanocolloid, four i.d. injections of 200 microCi/200 miccroL around the primary lesion) 24 hours before surgery and GP tracing during surgery. Subsequently ALND was performed for pathological assessment. RESULTS: SNs were identified in 73/76 patients using LS and in 72/76 using GP. In one case the SN was detected by GP alone while in two cases GP was not able to locate the SN although it had been identified by means of LS. Thirty-three of these 73 patients had axillary node involvement. In 31/33 cases the SN was the only positive node. No positive nodes were found in the remaining 40 ALNDs where SNs were identified. Thus, according to our experience 40/73 ALNDs could have been avoided. SNB seems to be a very interesting technique but further experience in lymph node radioisotope tracing is needed. PMID- 11016717 TI - Sentinel node and radioguided surgery in breast cancer of limited size. Preliminary experience at ASL 1, Massa e Carrara. AB - The aim of this study was to evaluate the effectiveness of sentinel node (SN) biopsy in breast cancer. Twenty-five female patients classified as T1N0 according to the TNM system of the UICC were evaluated with this procedure from April to October 1999. The day before surgery a subdermal injection of 99mTc-nanocoll within the tissue overlying the neoplastic lesion and subsequent lymphoscintigraphy were performed. In all patients the SN was detected with a radioguided probe during scintigraphy and surgery. Histological examination of the SN for detection of metastases was positive in four patients who subsequently underwent axillary dissection. In the remaining patients with normal SNs no axillary dissection was performed. The preliminary results confirm the validity of the sentinel node procedure. PMID- 11016718 TI - The sentinel lymph node in T1N0 breast cancer. AB - The authors report their experience gathered from December 1998 to December 1999 in the use of the sentinel lymph node (SN) method in breast cancer treatment. In 20 out of 21 cases (95%) localization of the SN was obtained by scintigraphy while in 19 cases (90.5%) the SN was found during surgery. Histological examination of the axillary lymph nodes gave a 95% accuracy with only one negative SN associated with positive axillary lymph nodes out of a total of 19. However, the authors have subjected all patients to a complete three-level axillary dissection since they believe the method applied has not yet been fully validated. PMID- 11016719 TI - Detection of sentinel node in breast cancer: pilot study with the imaging probe. AB - The commonly used gamma probes are easy to use but also give rough information when employed in radioisotope-guided surgery. When images are required for exact localization, a gamma camera as well as a probe have to be used. Position sensitive photomultipliers have contemporaneously allowed high-resolution scintigraphy and miniaturization of gamma cameras. We have assembled a miniature gamma camera with a 1-square-inch field of view and an intrinsic resolution of about 1 mm. When the minicamera is collimated with a large-holed, highly sensitive collimator, it acquires a spatial resolution of 3 mm. This prototype has been tested in the detection of difficult-to-image breast cancer sentinel nodes. Five nodes that had not been found with the usual technique of an Anger camera plus conventional probe were checked with the miniature camera that we named imaging probe: it actually is small enough to be used as a probe and large enough to give an image. One of the five nodes was found and imaged. It was small, disease-free, close to the tumor and probably hidden by the Compton halo around the peritumoral injection site. Our pilot study shows that the imaging probe, although still a prototype, has certain advantages over conventional methods when lymph node localization is required during surgery. PMID- 11016720 TI - Sentinel node biopsy for malignant melanoma: analysis of a four-year experience. AB - AIMS AND BACKGROUND: Sentinel node (SN) biopsy has been introduced to solve the controversy concerning the effectiveness of prophylactic lymphadenectomy in intermediate thickness melanoma. The aim of this study was to evaluate the rate of metastases, the technical details of the procedure, and the main reasons of failure. METHODS: 235 patients affected by intermediate thickness melanoma (tumor thickness >0.75 mm and <4 mm) without clinical signs of systemic spread (N0M0) were submitted to sentinel node biopsy between 1996 and 2000. Preoperative lymphoscintigraphy was routinely performed in the last 184 patients. Intraoperative mapping with gamma probe was combined with the use of vital dye for identification of sentinel nodes in the last 113 patients. RESULTS: The SN detection rate was 95.6%, with significant differences depending on the site of dissection and the use of a gamma probe. The overall rate of micrometastases was 14.7%, but relevant differences were recorded between different subgroups of patients (T2, 5.1%; T3a, 19.6%; T3b, 29%). CONCLUSIONS: Sentinel node biopsy requires a multidisciplinary approach (surgery, pathology and nuclear medicine) for reliable results. The association of vital dye and intraoperative gamma probe for sentinel node harvesting has made the procedure more effective, less time consuming and less invasive. Failures may be due not only to surgical mistakes, but also to improper nuclear medicine procedures or inaccurate histological evaluation of SNs. Methods for histological examination of the SN are still debated and not standardized but promising results have recently been obtained with molecular oncology techniques (RT-PCR). PMID- 11016721 TI - Detection of melanoma cells in peripheral blood and sentinel lymph nodes by RT PCR analysis: a comparative study with immunohistochemistry. AB - The presence of lymph node metastases is the best prognostic factor for predicting relapse or survival in melanoma patients. It has been demonstrated that melanoma metastases spread through the first lymph node(s) draining the tumor (sentinel lymph node, SN) to the lymphatic system and that detection of melanoma cells in peripheral blood directly correlates with prognosis in melanoma. To identify lymph node metastases and circulating melanocytes, we developed a single-step reverse transcriptase-polymerase chain reaction assay (RT PCR) for detection of two melanoma-specific markers: the tyrosinase gene, which encodes an enzyme associated with melanin synthesis, and melanoma antigen-related T-cells, which are present in tumor infiltrating T-lymphocytes. This method detects two tumor cells in a background of 10(7) lymphocytes. Thirty patients with stage I-IV cutaneous melanoma entered the study. Blood samples were taken preoperatively, one month after excision of the primary melanoma lesion and the SN or total lymphadenectomy, and before the start of chemotherapy and every three months thereafter in metastatic patients. SNs were collected from 22 patients, bisected and analyzed by RT-PCR and routine pathological and immunohistochemical tests. The preliminary results indicate that RT-PCR for melanoma markers is a sensitive and valuable method for the detection of micrometastases and for early diagnosis and staging of melanoma. PMID- 11016722 TI - Sentinel node biopsy in cutaneous melanoma patients: technical and clinical aspects. AB - The role of the patent blue dye (PBD) technique and intraoperative probe-guided lymphoscintigraphy (LS) in detecting the sentinel node (SN) was investigated in a group of 130 consecutive stage I cutaneous melanoma patients. The preoperative workup included high-resolution US scanning and LS performed 15-18 hours before surgery. On the basis of preoperative LS, in the group of examined patients a total of 143 lymphatic drainage basins were identified and surgically explored: 41.6% in the axilla, 52.8% in the groin, and 5.6% in the head/neck. A total of 228 SNs were intraoperatively detected and removed; 110 lymphatic basins contained histologically negative SNs, while 33 basins had metastatic SNs. The sensitivity for SN detection using PBD alone was 93%, while it was 100% when PBD was combined with intraoperative LS. Preoperative and intraoperative LS appears to be a highly sensitive technique for SN detection in cutaneous melanoma patients. Furthermore, in view of the limited skin incision when radioguided surgery is performed, SN biopsy could be feasible under local anesthesia. PMID- 11016724 TI - Mapping the sentinel lymph node in malignant melanoma by blue dye, lymphoscintigraphy and intraoperative gamma probe. AB - Eighty-eight consecutive patients (48 men and 40 women; mean age, 58.9 years; range, 16-84 years) with clinically localized cutaneous melanoma involving the trunk, extremities or head and neck underwent lymphatic mapping at our institution. The primary melanoma had a mean thickness of 2.74 mm (range, 0.95 to 9 mm). Patients were divided into two groups: group A (39 patients) underwent only vital blue dye (VBD) mapping, while group B (49 patients) underwent lymphatic mapping with VBD and radio-guided surgery (RGS) combined. In all patients 1-1.5 mL of VBD was injected subdermally around the biopsy scar 10-20 min before surgery. In group B 37 MBq in 150 microL of 99mTc-HSA nanocolloid was additionally injected intradermally 18 h before surgery (3-6 aliquots injected perilesionally). In all lymphatic basins where drainage was noted the sentinel lymph nodes (SNs) were identified and marked with a cutaneous marker. Final identification of the SN was then performed externally by a hand-held gamma probe. After the induction of anesthesia 0.5-1-0 mL of patent blue V dye was injected intradermally with a 25-gauge needle around the site of the primary melanoma. SNs were examined by routine hematoxylin and eosin (H&E) staining and immunohistochemistry. Patients with histologically positive SN(s) underwent standard lymph node dissection (SLND) in the involved lymph node basin. The SN was identified in 37/39 patients (94.9%) of group A and in 48/49 patients (98.0%) of group B. Blue dye mapping failed to identify the SN in 5 of the 88 patients (5.8%), while the radioisotope method failed in only 1 of 49 patients (2.0%). Similar results were obtained with the combined use of the two probes. The average number of SNs harvested was 1.9 per basin sampled, which does not differ significantly from the numbers reported by other authors. The SN was histologically positive in 18 patients (20.5%). None of the 12 patients with a Breslow thickness less than 1.5 mm had positive SNs, whereas 18 of the 77 patients (23.4%) with a Breslow index exceeding 1.5 mm showed metastatic SNs with H&E or immunohistochemistry. The latter all underwent SLND of the affected basin. In 10 patients (55.6%) the SN was the only site of tumor invasion; eight patients (44.4%) with positive SNs had one or more metastatic lymph nodes in the draining basin. PMID- 11016723 TI - Sentinel node biopsy in patients with cutaneous melanoma of the head and neck. AB - Biopsy of head and neck sentinel nodes (SNs) can be technically problematic due to the unpredictable and variable drainage patterns of this anatomic region. The aim of the present study was to evaluate the feasibility of SN biopsy for cutaneous melanoma of the head and neck. We performed SN biopsy in 17 patients affected by stage I cutaneous melanoma of the head and neck on the basis of lymphoscintigraphy, blue dye and gamma probe. A total of 24 procedures were performed. Drainage to more than one lymphatic basin was observed in five patients (two basins in three cases and three basins in two cases) and in all cases SN biopsy was performed in all basins. The biopsy distribution by site was: six cervical nodes, five parotid nodes, four supraclavicular and submandibular nodes, three auricular and axillary nodes. The SN identification rate was 87.5% (21/24); metastases were discovered in four cases, with a positivity rate of 23.6%. At the time of writing, 1 patient is alive with local disease, 3 patients are dead and 13 are alive and free of disease with a follow-up ranging from 1 to 40 months (median, 21 months) following SN biopsy. In our opinion preoperative lymphoscintigraphy and the intraoperative use of a gamma probe are useful for the identification of lymphatic drainage of cutaneous melanoma of the head and neck. PMID- 11016725 TI - Radioisotopic lymphatic mapping of the sentinel node in melanoma: importance of immunohistochemistry. AB - BACKGROUND: Elective lymph node dissection (ELND) for patients with malignant melanoma is still controversial. A possible alternative could be biopsy of the first tumor draining lymph node, the sentinel node (SN), which can be identified by means of radionuclide techniques. AIM: Our study was undertaken to assess the accuracy of lymph node biopsy and to stress the importance of immunohistochemistry (IHC) in the pathological assessment of the SN for improved staging of the primary tumor. METHODS: We performed lymphoscintigraphy (LS) in 183 melanoma patients (89 with melanoma of the legs, 11 of the arms and 83 of the trunk). Our protocol consisted of preoperative peritumoral i.d. injection of 99mTc-labeled microcolloid to define the regional lymphatic basin and identify the sentinel node by means of planar scintigraphy. In 147 of the 183 cases a gamma probe (GP) was used during surgery to trace the SN. Vital blue dye was used during surgery in all cases. The SNs were excised for pathological examination. The pathological status of the SN was defined by means of examination of frozen sections, hematoxylin-eosin staining and immunohistochemistry for S-100 and HMB 45 MAb. RESULTS: At least one separate focus of activity was identified by LS in 182 out of 183 patients; in all 147 cases where a GP was used, it was successful in tracing the SN. LS with cutaneous mapping of the SN successfully guided the surgical excision in 177 of the 183 cases; in the 7 remaining cases, i.e. 7 out of 83 cases with SNs in the axillary basin, GP was not used and no elective node dissection was performed. Metastases were found in 39 of these 177 cases. In all 39 cases the SNs were the only positive nodes in the basin. Of the 39 metastases 18 were identified by means of frozen section, 12 by means of hematoxylin-eosin, and 9 by means of immunohistochemistry. We therefore emphasize the importance of immunohistochemistry in the pathology of LS for improved staging of the primary tumor. PMID- 11016726 TI - Sentinel node biopsy in melanoma: the experience of Brescia Civic Hospital. AB - The accuracy of the sentinel node (SN) technique in the evaluation of lymph node involvement in melanoma was evaluated in 71 consecutive patients with localized disease and Breslow index >1 mm. Lymphoscintigraphy identified at least one SN in 70 of the 71 patients (98.5%). The following day 69 patients underwent selective SN excision. The SN was identified by portable probe. One hundred and twenty-two lymph nodes were removed, 14 (11.4%) of which were metastatic in 9 patients (13%). No metastases were found in 40 patients with Breslow <2 mm. Eight of the 9 patients with positive SNs underwent lymphadenectomy of the whole basin and in two patients new metastatic nodes were found. At 4-26 months' follow-up 1 of the 60 patients with negative SN (scalp melanoma with Breslow 6.2 mm) developed bilateral cervical metastatic nodes. Two more patients with Breslow 3.7 and 5 mm, respectively, developed liver and lung metastases. The remaining 57 patients are still disease free. Among the 9 patients with tumor-positive SNs, 1 was lost to follow-up, 3 died and 5 are still alive. Our data confirm the clinical reliability of the SN technique in melanoma; we feel the technique should be considered a standard tool in the evaluation of melanoma patients. PMID- 11016727 TI - Sentinel node biopsy in clinical stage 1 melanoma: rationale for restaging and follow-up. AB - Lymph node involvement appears to be the most significant prognostic factor in patients affected by melanoma and has been shown to reduce the five-year survival by 40%. We studied 31 patients (15 M; 16 F; age range, 28-83 years) with clinical stage 1 (CS1) intermediate thickness (0.75-4 mm) melanoma. Scintigraphic examination of the nodes was performed in all patients, 29 of whom underwent surgical biopsy of the SN after 24 hours. Early images were acquired 5, 15 and 79 min and late images 60-180 min following perilesional injection of 2-4 microdoses of 99mTc-nanocolloid (15-20 MBq). A cobalt marker was used to project the SN on the skin surface which was later stained with indelible ink. For intraoperative localization we used a portable probe and perilesional injection of patent blue violet dye, which proved positive in 24/29 patients (83%). After surgery histological examination of the sentinel lymph nodes (SNs) (hematoxylin-eosin and immunohistochemistry) found positivity for metastatic cells in 6 patients. They all underwent elective lymph node dissection (ELND); five are N0+ and are currently undergoing supportive therapy with interferon alpha with an 8-24-month follow-up, while one N+ patient died 14 months after surgery. Follow-up (3-26 months) of N0- patients has not evidenced any locoregional recurrence so far. Only one case showed hematogenic metastases. This procedure might radically change the therapeutic approach to CS1 melanoma because it is simple, scarcely invasive, and shows a favorable cost-benefit ratio. PMID- 11016729 TI - Primary anorectal malignant melanoma: report of two cases. AB - Primary anorectal malignant melanoma is a fairly uncommon but highly malignant disease. It is sometimes mistaken for benign conditions such as hemorrhoids or rectal polyps. Here we describe two cases of primary malignant melanoma of the rectum: in one patient a wide local excision (WLE) was performed and in the other an abdominoperineal resection (APR), both with curative intent. Both patients developed systemic recurrences and died of their disease at 24 and 10 months, respectively. In conclusion, the prognosis of anorectal melanoma is poor, irrespective of surgical treatment. WLE is the first choice for primary anorectal melanoma, while APR should be reserved for those cases where complete transrectal tumor resection is technically impossible. PMID- 11016728 TI - Sentinel node mapping for malignant melanoma. AB - Sixty-seven patients with malignant melanomas were seen in our institute since 1998. We looked for a sentinel node in 26 (38%) patients with lesions thicker than 1 mm, employing a double technique: staining with methylene blue and gamma probe mapping after injection of 99mTc. Forty-six nodes were retrieved, 25% of them containing tumor cells. This approach enabled us to choose between a wait and-see policy and radical surgery. PMID- 11016730 TI - Detection of sentinel nodes by lymphoscintigraphy and gamma probe guided surgery in vulvar neoplasia. AB - BACKGROUND: Pathologic lymph node status is the most important prognostic factor in vulvar cancer; however, complete inguinofemoral node dissection is associated with significant morbidity. Intraoperative lymphoscintigraphy associated with gamma detecting probe-guided surgery has proved to be reliable in the detection of sentinel node (SN) involvement in melanoma and breast cancer patients. The present study evaluates the feasibility of the surgical identification of inguinal sentinel nodes using lymphoscintigraphy and a gamma detecting probe in patients with early vulvar cancer. METHODS: Technetium-99-labeled colloid human albumin was administered perilesionally in 44 patients. Twenty patients had T1 and 23 had T2 invasive epidermoid vulvar cancer; one patient had a lower-third vaginal cancer. An intraoperative gamma detecting probe was used to identify SNs during surgery. Complete inguinofemoral node dissection was subsequently performed. SNs underwent separate pathologic evaluation. RESULTS: A total of 77 groins were dissected in 44 patients. SNs were identified in all the studied groins. Thirteen cases had positive nodes: the SN was positive in all of them; in 10 cases the SN was the only positive node. Thirty-one patients showed negative SNs: all of them were negative for lymph node metastasis. CONCLUSIONS: Lymphoscintigraphy and SN biopsy under gamma detecting probe guidance proved to be an easy and reliable method for detection of SNs in early vulvar cancer. If these preliminary data will be confirmed, the technique would represent a real progress towards less aggressive treatment in patients with vulvar cancer. PMID- 11016731 TI - Gamma probe-guided thoracoscopic surgery of small pulmonary nodules. AB - Video-assisted thoracic surgery (VATS) is an interesting and emerging procedure for the diagnosis and treatment of peripheral pulmonary nodules. We developed a new radioguided surgical technique for the detection during VATS of pulmonary nodules smaller than 2 cm, situated deep in the lung parenchyma and neither visible nor palpable with endoscopic instruments. The procedure is divided into two phases. Two hours before surgery 0.3 ml of a solution composed of 0.2 mL of 99mTc-labeled human serum albumin microspheres (5-10 MBq) and 0.1 mL of non-ionic contrast is injected into the lesion under CT guidance. Then the patient is submitted to VATS. During thoracoscopy a collimated probe of 11 mm diameter connected to a gamma ray detector is introduced via an 11.5 mm trocar and the pleural surface of the suspected area is scanned. A hot spot indicates the presence of the radiolabeled nodule and hence the area to be resected. We treated 39 patients with small pulmonary nodules (mean size, 8.3 mm; range, 4-19 mm). The patients were 27 men and 12 women (mean age, 60.8 years; range, 13-80 years). Nineteen patients had a history of synchronous or metachronous malignancy. In all cases the nodule was detected and resected and the resection margins were pathologically free of tumor. Histological examination showed 21 benign and 18 malignant lesions (7 metastases and 11 primary lung cancers). Nine patients with a frozen section-based histopathological diagnosis of lung cancer without functional contraindications underwent a completion lobectomy by open surgery in the same surgical session. In conclusion, the radiolocalization of small pulmonary nodules by gamma probe during VATS is a safe and easy procedure, with fewer complications and a lower failure rate than other localization techniques. PMID- 11016732 TI - Use of surgical gamma probe for the detection of lymph node metastases in differentiated thyroid cancer. AB - BACKGROUND: Patients with differentiated thyroid cancer (DTC) after total or near total thyroidectomy require 131I therapy. After surgery the persistence of lymph node metastases in our series of patients was frequent (30%). Such patients are preferentially treated with radioiodine and shifted to surgical reintervention when the nodal lesions persist after two 131I treatments. AIM: Use of an intraoperative radioactive probe (C-TraK) to allow a more radical surgical approach in thyroid cancer patients submitted to surgery for lymph node metastases. METHODS AND RESULTS: After adequate withdrawal of L-thyroxine suppressive therapy six patients were given high 131I doses followed by post therapy WBS which demonstrated cervical activity in 5 patients and peri-jugular activity in 1. Surgery with the help of a gamma probe allowed to detect and remove all metastatic nodes. After excision all surgical specimens showed higher radioactive counts with respect to the background. The post-surgical scan showed the disappearance of all areas of 131I uptake. Histology confirmed the presence of metastatic lesions from papillary thyroid cancer. CONCLUSIONS: We conclude that the use of a gamma probe can be successful in patients with metastatic neck lesions resistant to 131I treatment, particularly in patients with nonpalpable lesions. PMID- 11016733 TI - 99mTc-MIBI radioguided surgery for limited invasive parathyroidectomy. AB - We report the preliminary results obtained with the intraoperative MIBI-guided gamma probe in a group of 9 patients with primary hyperparathyroidism (pHPT). These patients were selected for limited invasive parathyroid surgery on the basis of a preoperative imaging protocol consisting of a pertechnetate & perchlorate/MIBI scan combined with neck ultrasonography (US). In the operating room 50-70 MBq MIBI was injected 30 to 45 min before parathyroidectomy. The radioactivity was measured intraoperatively at three sites: parathyroid (P), thyroid (T), and background (B). The P/B, P/T, and T/B ratios were calculated. The T/B ratio was relatively constant (range, 1.5-1.8; mean, 1.6), while a wide variability was observed both for P/T ratio (range, 1.2-2.3; mean, 1.7) and P/B ratio (range, 2.1-4.0; mean, 2.9). At surgery single enlarged parathyroid glands were easily identified by means of intraoperative MIBI-guided gamma probe. Moreover, the gamma probe allowed us to perform a limited 2-2.5 cm neck incision in eight patients affected by parathyroid adenoma. In the remaining patient a parathyroid carcinoma was diagnosed and a bilateral neck exploration was performed. The intraoperative MIBI gamma probe seems to be a useful aid when limited invasive parathyroid surgery is performed. PMID- 11016734 TI - Radioguided surgery of breast cancer: radiation protection survey. AB - The aim of this study was to estimate the radioactive risk for surgical staff performing radioguided sentinel lymph node (SN) biopsy and to calculate the contamination level in the operating room for assessment of the possible need for specific radiation protection procedures. We studied 20 patients who were selected for quadrantectomy and SN biopsy. The day before surgery a volume of 0.15 mL of 99mTc-nanocoll was injected: the activity was 3.11 +/- 0.85 MBq in group A (15 pts) and 11.6 +/- 0.6 MBq in group B (5 pts). External radiation to staff was evaluated by measuring the exposure rate in air one hour after radiopharmaceutical administration. The air KERMA rate during surgery was estimated considering the physical decay of 99mTc. Contamination of disposable materials and surgical instruments in the operating room was measured using a contamination monitor, whereas the residual activity in the SN and the injection site was measured with a gamma probe. The exposure rate at 20 cm from the injection site was 0.75 microSv/h when the most radioactive patients (group B) were treated. Contamination in the operating room proved to be negligible. Considering the number of radioguided treatments carried out by a surgeon in one year, an equivalent effective dose of 0.075 mSv was estimated; the recommended dose limit according to the relevant Italian law, DL 230/95, is 1 mSv/yr. Surgical staff therefore do not require a classification of "exposed workers" and there is no need to supply the operating room with special containers for radioactive waste. PMID- 11016735 TI - Surgical pathology of intramedullary spinal cord neoplasms. AB - The surgical pathology of intramedullary spinal cord neoplasms is most accurately based on radical resection specimens rather than on small biopsies, which may be highly misleading. A review of the neuropathology files at NYU Medical Center revealed 294 surgical specimens of intramedullary cord lesions examined between January 1, 1991 and December 31, 1998. Of these 117 were from children (age less than 21 years) and 177 were from adults (21 and over). While most types of central nervous system tumors known to occur in the brain also occur in the spinal cord, the different proportions of these tumors by histologic type, and the differences in the proportions of tumor types in children compared to adults, are both significant. In adults ependymomas are the predominant tumor type (93 total) while in children astrocytomas and mixed neuronal-glial tumors are virtually equally common and outnumber ependymomas. In this period no cord Primitive Neuroectodermal Tumors were identified. Among the astrocytic neoplasms and other gliomas, high grade tumors were distinctly uncommon in children and only slightly more common in adults, in sharp contrast with the brain, where the majority of adult intra-axial tumors are high grade. PMID- 11016736 TI - Magnetic resonance imaging of intramedullary spinal cord tumors. AB - The advent of MRI has significantly changed the diagnosis of spinal cord tumors. Standard imaging provides excellent localization and characterization of the tumor in a noninvasive fashion. Exact histologic diagnosis of the two most common tumors, ependymoma and astrocytoma, remains elusive but there are some suggestive imaging characteristics. It is hoped that some of the newer MR imaging sequences will improve characterization of the tumor and thereby influence therapy. Several of these pulse sequences are already used routinely in brain imaging. Evaluation of new imaging sequences in the spine has lagged behind brain MR imaging, mainly due to technical factors. Work on animal spine models and extrapolation from brain imaging studies suggest that it is only a matter of time before some of these techniques become clinically relevant. PMID- 11016738 TI - Spinal cord astrocytomas: presentation, management and outcome. AB - Intramedullary spinal cord astrocytomas are uncommon tumors. They are the most common spinal cord tumor in children and in adults are second only to ependymomas in frequency of occurrence. Low-grade histology predominates with high-grade lesions comprising only ten to fifteen percent of pediatric tumors and a slightly higher proportion in adults. Presenting symptoms typically evolve over months to years with regional back pain the most commonly reported initial complaint. Malignant tumors produce rapid neurological deterioration. MRI is the diagnostic modality of choice: spinal cord astrocytomas are iso- to slightly hypointense on T1, hyperintense on T2 and commonly have associated cysts. They enhance less intensely and are more eccentric than ependymomas. The goals of surgical intervention are to obtain a tissue diagnosis and resect as much tumor as possible without adversely affecting neurological function. Astrocytomas are infiltrating neoplasms and total resection is not generally possible. Somatosensory and motor evoked potential monitoring are routinely used but it is unclear if they improve outcomes. The operating microscope and bipolar cautery are essential surgical tools; the ultrasound and ultrasonic surgical aspirator are useful surgical adjuncts. Laminectomy is performed on adults while laminoplasty is favored for pediatric patients. Outcome for low-grade astrocytomas is less favorable than that of ependymomas with regard to both recurrence and function though many have prolonged survival. There is no correlation of extent of resection and recurrence. Outcome for high-grade tumors is extremely poor; tumor progression is relentless; median survival is thirteen months in children and six months in adults. PMID- 11016737 TI - Intramedullary ependymomas: clinical presentation, surgical treatment strategies and prognosis. AB - Intramedullary ependymomas are rare tumors but comprise the majority of intramedullary glial neoplasms in the adult. These tumors are benign, slow growing lesions which are optimally treated with gross-total surgical resection without adjuvant therapy. This objective can be attained safely in a majority, of patients. Post-operative functional outcome is related to pre-operative functional status. Hence, early diagnosis, prior to symptomatic progression, is critical to the successful treatment of these tumors. Adjuvant therapy is indicated for the rare malignant or disseminated tumor or following sub-total resection. PMID- 11016739 TI - Pediatric intramedullary spinal cord tumors: special considerations. AB - Intramedullary spinal cord tumors (IMSCTs) of the pediatric population are rare and comprise thirty-five percent of intraspinal neoplasms. Low-grade astrocytomas predominate; ependymomas increase in frequency with ascending age and become the most frequent IMSCT in adults. Gangliogliomas are very rare in adults but comprise nearly thirty percent of tumors in children under three years of age. The cervical spine is the region of the spine most affected. Pain is the most common presenting symptom with weakness, gait deterioration, torticollis also frequently reported. Hydrocephalus occurs with greater frequency than in adult patients and often requires a shunt. Motor and sensory evoked potential monitoring is routinely utilized. Osteoplastic laminotomy is performed to forestall the development of progressive spinal deformity. Gross total resection is feasible in most ependymomas and results in surgical cure. Astrocytomas are infiltrating neoplasms and gross total resection is occasionally possible only in the pediatric population. However, the role of radical resection of low-grade fibrillary astrocytomas of the spinal cord in children has not been definitively demonstrated in the literature. Outcome for low-grade astrocytomas is better in children than adults, but not as favorable as that for ependymomas. Malignant tumors have dismal outcomes and surgery in these patients serves only to provide a diagnosis. PMID- 11016740 TI - Radiation therapy and the management of intramedullary spinal cord tumors. AB - The use of radiation therapy in the management of intramedullary spinal cord tumors remains controversial. Several studies indicate that the use of postoperative radiation therapy modestly improves both local control and survival in spinal cord ependymomas and astrocytomas. Modern treatment planning and imaging allow more accurate target definition and respect for related normal tissue tolerances. PMID- 11016741 TI - Genetic basis of intramedullary spinal cord tumors and therapeutic implications. AB - The advent of molecular biology has provided tools to delineate genetic mutations that cause disease. Recently, several genetic mutations have been associated with intramedullary spinal cord tumors. Concurrently, advances in micro-neurosurgical techniques have significantly decreased the morbidity of surgical resection. In this review, we describe the current understanding of genetic mutations in sporadic and familial intramedullary spinal cord tumors. The future success of innovative gene therapy protocols may depend upon establishing a cause and effect relationship between these genetic mutations and disease progression. Successful gene therapy will also depend upon increasing the efficiency of gene therapy vector delivery. PMID- 11016743 TI - Intramedullary spinal tumors of disordered embryogenesis. AB - Abnormal spinal embryogenesis is quite commonplace. While greater than 90 percent of these errors of embryogenesis leads to occult spinal dysraphism with minimal neurologic or orthopedic sequelae, there is a significant minority of these anomalies which leads to the formation of the so-called 'congenital tumors of disordered embryogenesis'. The purpose of this article is to discuss the embryology, presentation, diagnosis and management of the spinal dysraphic states with particular emphasis on those errors which lead to mass lesions in the spinal canal such as dermoids, epidermoids, lipoma/lipomyelomeningocoele and neurenteric cysts. We also include lesions such as dermal sinus tracts and thickened filum terminale in our discussion with particular emphasis on their relationship to the tethered cord syndrome. Proper surgical management of these various conditions necessitates a thorough understanding of their embryologic etiology and the anatomic/physiologic ramifications that such lesions have on the developing spinal cord. PMID- 11016744 TI - Non-neoplastic intramedullary pathology. Diagnostic dilemma: to Bx or not to Bx. AB - There are several non-neoplastic lesions which may mimic intramedullary spinal cord neoplasm in their radiographic and clinical presentation. These can be classified as either infectious (TB, fungal, bacterial, parasitic, syphilis, CMV, HSV) and non-infectious (sarcoid, MS, myelitis, ADEM, SLE) inflammatory lesions, idiopathic necrotizing myelopathy, unusual vascular lesions (amyloid, infarct, isolated intramedullary vascular lesions) and radiation myelopathy. Although biopsy may be indicated in many cases, the mistaken diagnosis of intramedullary neoplasm can often be eliminated pre-operatively. PMID- 11016745 TI - Chemotherapy for intramedullary spinal cord tumors. AB - Intramedullary tumors are rare, accounting for only about 4% of all CNS neoplasms. Although surgery represents the most effective treatment, recurrence may occur. As a large proportion of intramedullary malignancies occur in children, who are more sensitive to the deleterious effects of irradiation, chemotherapy assumes an important role. This article describes the most common intramedullary tumors and the role of chemotherapy. PMID- 11016742 TI - Hemangioblastomas and other uncommon intramedullary tumors. AB - Astrocytoma and ependymoma make up 90% of intramedullary tumors between them. However, a host of less common tumors form the remaining 10%: these include hemangioblastomas, subependymomas, gangliogliomas and other neuronal variants, metastases from extraneural cancers, and a host of other lesions that typically occur intracranially but which present on rare occasions in the intramedullary location. Most neurosurgeons will encounter the unusual tumors of the spinal cord described in this review only a few times during their professional careers, but it is nevertheless important to recognize the distinct radiological and intraoperative features of those for which significant series of patients have been accumulated. Metastases and germinomas aside, the other neoplasms described here are relatively benign in their clinical and histological behavior, and can be meaningfully resected by careful microsurgical technique. PMID- 11016746 TI - Cisplatin-epirubicin-paclitaxel weekly administration with G-CSF support in advanced breast cancer. A Southern Italy Cooperative Oncology Group (SICOG) phase II study. AB - PURPOSE: It has been shown in vitro that both cisplatin and epirubicin increase the antitumor activity of paclitaxel. Weekly administration could give a substantial improvement in the therapeutic index of cisplatin and paclitaxel. This study was aimed at defining the antitumor activity of a weekly cisplatin epirubicin-paclitaxel (PET) administration in locally advanced or metastatic breast cancer patients. PATIENTS AND METHODS: Sixty-eight breast cancer patients with advanced disease, who had not received prior chemotherapy (except adjuvant), received weekly cisplatin 30 mg/sqm, paclitaxel 120 mg/sqm and epirubicin 50 mg/sqm plus G-CSF (day 3-5), for a maximum of 12 cycles. Thirty-five patients had stage IIIB and 33 stage IV disease (14 with visceral metastases). RESULTS: All patients were evaluable for response on an intent to treat basis. Overall, 21 complete and 38 partial responses have been recorded for an 87% ORR (95% CI = 76 94%). Fourteen CRs and 19 PRs have been registered in the 35 patients with locally advanced disease for a 94% ORR (95% CI = 81-99%) while 7 CRs and 19 PRs were observed in the 33 patients with metastatic disease for a 79% ORR (95% CI-61 91%). Surgery was performed in 33/35 women with locally advanced disease. Four of these patients (11%) showed no invasive cancer on pathologic examination, and in an additional 8 patients tumor < 1 cm was found in the breast. Only 4/33 patients who underwent surgery relapsed. The projected one-year RFS was greater than 80%. At an 11-month median follow-up (range, 3-19), 11 patients had progressed and 5 had died among the 33 patients with metastatic disease, the median progression free survival in this group being 14 months. Severe hematologic toxicity was uncommon, grade 3-4 neutropenia and thrombocytopenia occurring in 32% and 4% of patients, respectively. Only 2 episodes of neutropenic sepsis were registered. Packed red blood cell transfusions were required in 7 patients. Vomiting, diarrhoea, mucositis and skin toxicity were severe in 6%, 9%, 10%, and 9% of patients, respectively. Peripheral neuropathy was observed in 47% of patients. CONCLUSIONS: The weekly PET administration is a well tolerated and very effective approach in advanced breast cancer patients. It can produce a 40% clinical complete response rate, with a more than 10% pCR rate in patients with T4 disease, and an about 80% ORR in those with distant metastases. A phase III trial comparing PET with a standard every 3 weeks epirubicin-taxol administration is underway. PMID- 11016747 TI - Transcriptional suppression of synuclein gamma (SNCG) expression in human breast cancer cells by the growth inhibitory cytokine oncostatin M. AB - Previously, we have shown that synuclein gamma (SNCG), a member of the brain protein synuclein family, is highly expressed in human infiltrating breast carcinomas but not expressed in normal or benign breast tissues. The SNCG mRNA was also detected in several human breast cancer cell lines with the highest expression found in H3922, a cell line derived from an infiltrating ductal carcinoma. In this study, we show that expression of SNCG mRNA in H3922 cells is significantly decreased by treating cells with the cytokine oncostatin M (OM) who has a growth-inhibitory effect on these cells. A decrease in SNCG mRNA level can be detected as early as 30 min after OM addition. By 4 h OM treatment, the level of SNCG mRNA was decreased to 70% of control, and by 24 h the mRNA was below detectable level. Western blot analysis further demonstrated the suppression of SNCG protein expression by OM. The level of SNCG protein in H3922 cells was reduced more than 90% by OM after 2 days. Since OM-induced growth inhibition occurs after 3-4 days, the down-regulation of SNCG expression appears to proceed the effect of OM on cell growth. Additional experiments to measure the transcriptional rates of SNCG indicate that the observed OM-induced down regulation of SNCG mRNA occurs mainly at the transcriptional level. In an attempt to examine the role of SNCG gene in the proliferation of breast cancer cells, SNCG cDNA was stably transfected into MCF-7 cells that do not express endogenous SNCG gene. Examination of cell growth under anchorage-dependent and anchorage independent conditions demonstrates that over expression of SNCG gene significantly stimulated the growth of MCF-7 cells both in monolayer culture and in soft agar. These data together suggest that SNCG may play a role in cell proliferation. PMID- 11016748 TI - Inducible expression of herpes simplex virus thymidine kinase increases sensitivity to ganciclovir but does not enhance bystander effect in breast cancer cells. AB - Delivery of cancer chemotherapy directly to the cancer cell has great appeal. Previous studies using adenoviral transfer of the herpes simplex virus thymidine kinase (HSV-tk) gene followed by ganciclovir (GCV) in an ascites model of breast cancer was successful in reducing tumor burden and prolonging life. However, increasing the viral dose resulted in increased toxicity and host mortality emphasizing the need for an improved therapeutic ratio. To test the hypothesis that enhancement of HSV-tk gene expression would lead to increased sensitivity to GCV and improved bystander effect, we created breast cancer cells expressing HSV tk under the control of the inducible tetracycline promoter. Using this system, we could inducibly increase gene expression and biochemical activation of HSV-tk. These increased levels of HSV-tk decreased the IC50 to GCV nearly 50-fold. However, the bystander effect was not enhanced by increasing HSV-tk gene expression. We conclude that increased HSV-tk gene expression improves sensitivity to CCV. However, additional measures, such as increased gap junction communication, will likely be needed to enhance the bystander effect and the therapeutic efficacy of this strategy. PMID- 11016749 TI - Flow cytometry in primary breast carcinomas. Prognostic impact of proliferative activity. AB - From 1990 to 1996, 607 previously untreated, node-negative, invasive breast carcinomas were sampled by a pathologist for flow-cytometric DNA analysis. The aim of the present work was to study the correlations between flow cytometric results obtained thanks to the American Consensus (AC) guidelines of 1993 and the established clinico-pathological prognostic factors (T, grade, receptors), and despite a short global follow-up (mean of 4 years), to correlate flow cytometry with the outcome of the patients. In this study S-phase fraction (SPF) correlated strongly with tumor size, histological grade, lack of steroid receptors, histological type and was together with the mitotic activity a paramount prognostic factor even after multivariate analysis. This study compared also the technical criteria proposed by the AC with our own more stringent ones and concluded that the criteria of the AC are relevant and allow, thanks to the use of tertiles in the reporting of SPF values, a comparison of values obtained by different teams. Our review of the literature, focused on series using fresh material, enabled us to show that there is a rather wide agreement concerning the relationship between SPF and prognosis most often after multivariate analysis. This despite the lack of standardization in the design of the studies (implementation of the technical steps or reporting of results). When estimated from fresh or frozen material following AC's guidelines. SPF along with mitotic activity should become a prognostic factor used in the daily practice by oncologists in the management of breast carcinomas. PMID- 11016750 TI - A phase II trial of cyclophosphamide, epirubicin and vinorelbine in the treatment of advanced breast cancer. AB - BACKGROUND: Vinorelbine (Navelbin; N) has proven to be active in patients with advanced breast cancer (ABC) and cyclophosphamide (C) and epirubicin (Epiadriamycin: E) are still among the main cytostatic agents against this tumor. On this basis was carried out a study to determine the activity and toxicity of the combination of these three agents (CEN). PATIENTS AND METHOD: From April 1996 to March 1998, 59 patients with ABC were recruited of whom 56 were found eligible and evaluable for toxicity and 55 for activity. The treatment regimen was C: 400 mg/m2, E: 30 mg/m2 and N: 25 mg/m2 administered intravenously on days 1 and 8 of a 28-day cycle. RESULTS: The median number of cycles administered was 6 (range: 1 16). The most common hematological toxicity was grade (G) 3 and 4 neutropenia occurring in 36% of patients, associated with fever in 7% of them. Grade 3-4 thrombocytopenia and anemia occurred in 5% and 7%, respectively. Other G2-G3 non hematologic toxicities were: N/vomiting in 34%, alopecia in 73% and mucositis in 11% of patients. An objective response was achieved in 28 of 56 patients (50%) (95% confidence interval (CI): 37-63%): complete response (CR) in 9%, partial response (PR) in 41%. The median duration of response, time to progression and overall survival time was 54, 47 and 90 weeks, respectively. CONCLUSION: The CEN combination at these doses and treatment schedule appears to have acceptable tolerability but there is no apparent improvement in therapeutic efficacy when compared to other regimens used as first line treatment in ABC. PMID- 11016751 TI - Five-day infusion fluorouracil plus vinorelbine in women with breast cancer previously treated with anthracyclines and paclitaxel. AB - The continuous infusion of fluorouracil presents a superior pharmacological profile than its bolus administration, while vinorelbine is a new drug associated with good clinical activity in pretreated metastatic breast cancer. We investigated the combination of this two antitumor drugs with the aim to determine a tolerant and active second-line therapy in metastatic pretreated patients. PATIENTS AND METHODS: Fifty six patients pretreated with chemotherapy received a median of six cycles [2-11] of fluorouracil, 700 mg/m2 for 5 day continuous i.v. infusion and vinorelbine, 20 mg/m2 on days 1 and 6, every three weeks. The inclusion and evaluation criteria required measurable disease by conventional clinical and/or instrumental means. FINDINGS: Iatrogenic toxicity in 340 administered cycles was mild: stomatitis = 11% (Grade 3 = 5%), constipation and abdominal pain = 12%, G2 neutropenia = 4%, G1 thrombocytopenia = 0.5%. In nine cases moderate infections occurred and six women experienced catheter related complications. Complete and partial remissions were observed in 12% and 36% of evaluable patients, respectively. In particular major tumor regression was documented in 28% of anthracyclines or taxol unresponsive cases. CONCLUSIONS: This drug combination is active in metastatic pretreated breast cancer patients and devoid of serious iatrogenic toxicity. Although it deserves future optimization, for instance with the inclusion of oral fluoropirimidines, it represents a good choice for second-line or non cross-resistant regimens. PMID- 11016752 TI - Quality of life after adjuvant chemotherapy for breast cancer. AB - PURPOSE: To evaluate the quality of life of breast cancer patients previously treated with adjuvant chemotherapy. METHOD: Registry data were used to recruit a sample of breast cancer patients (N = 61; mean age = 51.6 years) with no current evidence of disease who had completed adjuvant chemotherapy between 3 and 36 months earlier (average = 15.87 months). In addition, a peer nomination procedure was used to recruit an age-matched comparison group of women with no history of cancer (N = 59; mean age = 51.5 years). Both groups were mailed a survey to complete that included the Medical Outcomes Study Short Form 36 (SF-36) and the Center for Epidemiologic Studies Depression Scale (CES-D). These data were used to test the hypothesis that breast cancer patients previously treated with adjuvant chemotherapy experience impaired quality of life relative to their peers and to identify demographic and medical factors associated with individual differences in patient quality of life. RESULTS: Consistent with predictions, the postchemotherapy group scored poorer than the noncancer comparison group on the CES-D and on six of the eight subscales as well as the physical component summary scale of the SF-36 (p < 0.05). With regard to individual differences in patient quality of life, younger age and unmarried status were positively related to poorer mental well-being and greater depressive symptomatology (p < 0.05). Time since cancer diagnosis and chemotherapy completion were also positively related to greater depressive symptomatology (p < 0.05). In contrast, none of the demographic or medical variables assessed were related to physical well-being (p > 0.05). CONCLUSIONS: Breast cancer patients appear to experience problems in multiple quality of life domains following the completion of adjuvant chemotherapy treatment. Demographic and medical characteristics explain individual differences in mental but not physical aspects of patient quality of life. These findings demonstrate the need for interventions to improve the quality of life in breast cancer patients previously treated with adjuvant chemotherapy. PMID- 11016753 TI - Characterization of a novel silencer element in the human aromatase gene PII promoter. AB - Approximately two thirds of breast cancer patients have estrogen-dependent carcinomas. The biosynthesis of estrogens is catalyzed by the microsomal enzyme aromatase. Mechanisms controlling human aromatase gene expression are complicated by the existence of multiple tissue specific promoters. The most proximally located Pll promoter is mainly active in ovarian granulosa cells. PlI can be switched on in human breast cancer cells. Since there are strong silencer elements located within the 3' portion of the PlI promoter, we propose that the function of these silencer elements could be reversed by breast cancer cell specific signals/factors, resulting in aberrant expression of aromatase. We have identified and characterized a novel silencer element, S2, which is upstream of S1, a silencer element recently identified by another group. S2, a 54-bp fragment 100% conserved between humans and rodents, functions in both orientation- and promoter-independent manners. The core region of S2 contains two consensus binding sites for members of the GATA transcription factors. GATA-4 was found to be expressed in three out of four human breast cancer cell lines examined by RT PCR, and transfection with GATA-4 partially reversed the repressive function of S2. However, we were unable to demonstrate that DNA-protein complexes formed between nuclear extracts of human breast and ovarian cancer cells and S2 contain GATA-4 using a supershifting approach. We suggest that the expression of GATA-4, and more importantly, other yet to be identified GATA or GATA-related factor(s), are implicated in provoking aberrant expression of aromatase, and therefore, the biosynthesis of estrogens, in human breast cancer cells. PMID- 11016754 TI - Acute neuromedical and neurosurgical admissions. Standard and ultrafast MR imaging of the brain compared with cranial CT. AB - PURPOSE: To evaluate the role of standard and ultrafast MR brain imaging and compare the information with CT. MATERIAL AND METHODS: This was a prospective study of 114 patients with acute neurological symptoms and signs. CT brain examinations consisted of axial non-enhanced images. MR imaging consisted of standard spin-echo/fast spin-echo sequences and a series of rapid techniques including echoplanar and single shot fast spin-echo sequences. RESULTS: Using standard MR methods, 41% of the patients had all five sequences of good technical quality, while using ultrafast methods 81% of the patients had good technical quality examinations in all five sequences. In 3% of the cases, ischaemic stroke was incorrectly reported on CT. In 24% of the cases, MR gave extra diagnostic information not reported on CT and in a further 8%, one neuroradiologist reported the abnormality in agreement with the MR, whilst the other neuroradiologist reported the CT as normal. In 2 cases, subarachnoid haemorrhage was missed on MR. Subarachnoid haemorrhage was not shown on the ultrafast sequences. CONCLUSION: MR can be used to image acute neurological admissions with a high success rate, particularly using ultrafast methods. In many cases, MR provided extra information of direct clinical relevance not shown on CT. PMID- 11016755 TI - Persistence of both proatlantal arteries with occlusion of the internal carotid arteries. A case report. AB - A case of bilateral persistence of the proatlantal arteries with occlusion of both internal carotid arteries is reported. PMID- 11016757 TI - Imaging features of myoepithelioma arising from the hard palate. A case report. AB - A 53-year-old Japanese woman presented with a mass of the hard palate that was histologically diagnosed as myoepithelioma. By CT, the well-demarcated ovoid isodense mass showed faint contrast enhancement. By MR imaging, the mass was homogeneous and isointense on T1-weighted images with homogeneous enhancement after administration of contrast material and markedly hyperintense on T2 weighted images. Myoepithelioma should be kept in mind as differential diagnosis of hard palatine tumors. PMID- 11016756 TI - Contrast-enhanced color Doppler ultrasonography in suspected parathyroid lesions. AB - PURPOSE: To assess the usefulness of US contrast media in the evaluation of patients with primary hyperparathyroidism, with a nodule showing US features of a primary parathyroid lesion but lacking the color Doppler US appearance of a parathyroid mass. MATERIAL AND METHODS: Thirteen patients (7 female, 6 male; age range 51-79 years) were examined with US before and after administration of a stabilized galactose-based microbubble contrast agent. Ten patients underwent surgery and the final histological examination demonstrated parathyroid adenoma in 9 cases and a mesenchymal benign nodule in 1 case. Three nodules were proved to be of thyroid origin at fine-needle aspiration biopsy. RESULTS AND CONCLUSION: The use of a US contrast agent resulted in a diagnostic gain compared to unenhanced studies in 12/13 cases. Color Doppler findings characteristic of parathyroid lesions were observed in 7/13 cases, of thyroid nodules in 4/13 cases, and nonspecific patterns in 2/13 cases. Contrast-enhanced color Doppler US can be proposed in selected patients in whom unenhanced color Doppler provides uncertain findings. Its ideal application should be the evaluation of cervical lesions without detectable intranodular flow at unenhanced Doppler studies. In these cases, the contrast agent helps in visualizing typical color Doppler signals of the parathyroid lesions ("vascular pole" and "mixed pattern"). PMID- 11016758 TI - Hard-copy versus soft-copy with and without simple image manipulation for detection of pulmonary nodules and masses. AB - PURPOSE: To compare interpretation performance on soft-copy presentations, with and without simple image manipulation, and on unmodified hard-copy presentations with regard to detection of pulmonary nodules and masses. MATERIAL AND METHODS: Fifty chest digital radiograph combinations of patients with a total of 60 nodules, 32 of which were 2.0 cm in diameter, were selected for the study. Three readers evaluated three separate image formats: unmodified hard- and soft-copies, and soft-copies with simple image manipulation of lung and mediastinum window settings, and zooming. The screen display was 1600 x 1200 pixels with 8 bits/pixel. RESULTS: The sensitivity, accuracy, detectability, and Az value of the soft-copy systems were clearly inferior to hard-copy evaluation. The mean Az values were 0.921 for unmodified hard-copy, 0.820 for image-manipulated soft copy, and 0.781 for unmodified soft-copy. CONCLUSION: Soft-copy interpretations were not as sensitive in detecting pulmonary nodules and masses as hard-copy evaluation. PMID- 11016759 TI - Symptomatic improvement in dyspnea following tracheobronchial metallic stenting for malignant airway obstruction. AB - PURPOSE: To investigate the value and limitation of Gianturco expandable metallic stenting for patients with dyspnea due to stenotic tracheobronchial lesions associated with malignancies. MATERIAL AND METHODS: We treated 55 lesions of 44 patients with obstructing stenotic tracheobronchial lesions related to end-stage malignancies by Gianturco expandable metallic stents (EMSs). RESULTS: In 42 of 44 patients, the dyspnea subjectively improved after the procedure (95.5%). An improvement over one grade of the Hugh-Jones classification was shown in 79.5% (35/44); in 80% (20 of 25 patients) with intraluminal tumor and in 78.9% (15 of 19 patients) with extrinsic compression. Seven of the 44 patients developed dyspnea related to re-stenosis of 10 lesions and 1 of these patients developed dyspnea related to re-re-stenosis during follow-up. The mean duration of survival was 4.3 months in patients who underwent stenting. No significant differences in survival rates and primary patency rates were seen in patients with extrinsic compression compared to patients with intraluminal tumors. CONCLUSION: Gianturco EMS therapy was valuable in patients who suffered from dyspnea due to airway stenosis causing obstruction. PMID- 11016760 TI - CT features of mediastinal lymphangiohemangioma associated with superior vena cava ectasia. A case report. AB - We present a case of mediastinal mesenchymal tumor in a 53-year-old woman with a 1-month history of chest tightness. A histological diagnosis of lymphangiohemangioma was obtained by excisional biopsy. CT findings are reported with emphasis on its association with intratumoral superior vena cava ectasia. PMID- 11016761 TI - Poland's syndrome with lung cancer. A case report. AB - Poland's syndrome, a rare congenital anomaly characterized by pectoralis muscle defect, has been reported in association with lymphoreticular malignancies and some solid tumors. Lung cancer associated with Poland's syndrome has not been previously described. We present the first report of a case of Poland's syndrome associated with lung cancer and demonstrate the CT findings. PMID- 11016762 TI - Local breast cancer recurrence caused by mammographically guided punctures. AB - PURPOSE: To evaluate the risk of needle track seeding or tumor cell implantation as the cause of locally recurrent breast cancer after breast conserving surgery. MATERIAL AND METHODS: We reviewed recurrences from a consecutive series of 303 clinically nonpalpable breast cancers treated with breast conserving surgery after pre-operative localization. The median mammographic follow-up was 5.4 years. The suspicion of seeding or implantation was based on the location of the recurrent lesion in comparison with the needle path in two orthogonal mammographic projections. Pre-operative percutaneous biopsies had been done in 71% (214/303) of the cases. Postoperative radiotherapy was administered to 82% (194/238) of the invasive cancers and to 28% (18/65) of the ductal cancers in situ (DCIS). RESULTS: Locally recurrent cancer occurred in 11% (33/303) of the cases. Radiotherapy demonstrated a protective effect from relapse among invasive cancers but not for DCIS. Seeding or implantation was suspected in 3 recurrent invasive cancers which had not been subject to radiotherapy. The histopathological diagnosis of the primary cancer and the recurrent cancer were the same in these cases: adenoid cystic, mucinous and tubuloductal cancer. CONCLUSION: Seeding or implantation was suspected as the cause of local recurrence in 7% (3/44) of the invasive cancers which did not receive radiotherapy. PMID- 11016763 TI - The role of scintimammography with 99mTc-sestamibi as a complementary diagnostic technique in the detection of breast cancer. AB - PURPOSE: To prospectively determine the clinical value of scintimammography (Sc) with 99mTc-sestamibi as a complementary method to triple diagnosis (TD) in detecting malignant disease of the breast. MATERIAL AND METHODS: Ninety-six patients with 119 clinically or mammographically detected breast lesions underwent TD procedures, including clinical examination, mammography and fine needle aspiration cytology. Prone planar Sc with 99mTc-sestamibi was performed in all 96 patients. Five groups were defined for diagnosis: 1=normal; 2=benign; 3=probably benign; 4=highly suspect of malignancy; and 5=malignant. All lesions were histopathologically examined. The results of each method per se and the combination of TD with Sc (TD+Sc) were analyzed. RESULTS: Histopathology of the 119 surgically excised breast lesions found 83 malignant and 36 benign lesions. TD missed 6 of 83 carcinomas, resulting in a sensitivity of 92.7%. Sc alone showed sensitivity of 85.5%. The combination TD+Sc missed 1 of 83 carcinomas, and thus had a sensitivity of 98.7%. In mammographically dense breasts both TD and Sc detected 16 of 18 carcinomas, while the combination TD+Sc led to detection of all 18 carcinomas. CONCLUSION: Adding Sc to TD increases the sensitivity for detection of breast carcinomas. Sc with 99mTc-sestamibi is recommended as a complimentary method to TD in selected cases such as mammographically dense breasts. PMID- 11016764 TI - Fluor-18-fluorodeoxyglucose positron emission tomography (FDG-PET) in malignant melanoma. Diagnostic comparison with conventional imaging methods. AB - PURPOSE: To assess the diagnostic value of fluor-18-fluorodeoxyglucose positron emission tomography (FDG-PET) in screening for melanoma metastases. MATERIAL AND METHODS: The case records of 94 melanoma patients who had been examined by whole body FDG-PET between 1995 and 1999 were evaluated retrospectively. Forty patients showed evidence of lymphogenous and 42 of hematogenous metastasis. The maximal interval between PET and the diagnostic procedure under comparison was 2 weeks. Confirmation of the findings was based on histology or the clinical or radiological course. RESULTS: In 24 patients, all diagnostic examinations including CT had been performed within 2 weeks from PET. In no case did PET change the staging. In 13 patients, PET agreed with morphological diagnosis in the number of metastatically invaded organs. This included 3 patients without metastases. The estimated number of organs invaded by metastases was higher with PET in 5 patients and higher with morphological imaging techniques in 6 patients. Among the PET findings with higher or equivocal counts of organs with metastases there were 2 confirmed false-positive findings. CONCLUSION: In a selected patient population, FDG-PET was found to be inferior to CT for diagnosing lung and liver metastases. The supplementary use of FDG-PET is not generally of value once metastasis has been established. PMID- 11016765 TI - Differentiation of vascular birthmarks by MR imaging. An investigation of hemangiomas, venous and lymphatic malformations. AB - PURPOSE: To evaluate the use of MR imaging using standard sequences in the differentiation of common vascular birthmarks of soft tissues in childhood. MATERIAL AND METHODS: Forty-three lesions in 37 children (median age 33 months) with hemangiomas (n=25), venous (n=8) and lymphatic (n=10) malformations were retrospectively studied. Images were obtained with a 0.2 T and a 1.5 T MR unit, using T1-weighted spin-echo, T2-weighted turbo spin-echo and turbo STIR sequences. In addition, 19 contrast studies were evaluated. Signal intensities and morphological characteristics were analyzed. RESULTS: All hemangiomas were hyperintense in T2-weighted, and mostly (22/25) isointense compared to the surrounding skeletal muscle in T1-weighted images. The lesions were homogeneous or mildly inhomogeneous and well defined. Venous and lymphatic malformations showed similar patterns. Contrary to other lesions, no vessels or lobular architecture were detectable and contrast enhancement was absent in lymphatic malformations. CONCLUSION: A reliable differentiation between hemangiomas, venous and lymphatic malformations in childhood is not possible by standard MR sequences alone. Contrast media may be helpful as lymphatic malformations show no contrast enhancement. The main role of MR imaging is to determine the local extent or infiltration of deeper tissue layers and narrow the differential diagnosis of other soft tissue tumors. PMID- 11016766 TI - High-dose percutaneous ethanol injection therapy of liver tumors. Patient acceptance and complications. AB - PURPOSE: To study the safety of high-dose ethanol injections in liver tumors and their acceptability as out-patient procedures under local anesthesia. MATERIAL AND METHODS: Twenty-nine patients received 20-95 ml of 95% ethanol by 62 percutaneous injections under local anesthesia. Pain was assessed by a visual analogue scale from 1 to 10 for 33 of the sessions. Side effects, complications, post-treatment requirement of parenteral analgesics, and hospital stay (discharge the same day or later) were recorded. The sessions were compared to 80 injections of <20 ml of ethanol in 18 patients. RESULTS: High-dose injections with an average volume of 39 ml gave a mean pain score of 5.1, with a weak relationship between pain and volume. Other side effects and complications were unrelated to the ethanol dose. They comprised 1 syncopation, 1 occasion of hypoventilation requiring antidote to opiates, 12 short episodes of nausea or vomiting without need for i.v. fluids, 2 instances of sepsis, and 1 abscess that was drained percutaneously. Thirty-nine of the 62 sessions were performed in day care. Ethanol was given in high doses without apparent complications after or shortly before liver resections (3 patients in each group), and on 4 occasions in 2 HIV carriers. Low-dose injections resulted in a mean pain score of 4.7 with the same requirement of i.v. analgesics as high doses, fewer instances of nausea and no infectious complications. CONCLUSION: High-dose ethanol injections in patients with liver malignancy had no mortality and a reasonable complication rate. They could be given without general anesthesia, often in day care. PMID- 11016767 TI - Diagnostic accuracy of helical CT arterial portography and CT hepatic arteriography for hypervascular hepatocellular carcinoma in chronic liver damage. An ROC analysis. AB - PURPOSE: To evaluate the detectability of hypervascular hepatocellular carcinomas (HCCs) in chronic liver damage with helical CT arterial portography (CTAP) and CT hepatic arteriography (CTHA). MATERIAL AND METHODS: Thirty-nine HCC patients who underwent CTAP and CTHA were studied. Diagnostic abilities of CTAP alone, CTHA alone, or combined CTAP and CTHA were evaluated by receiver operating characteristic (ROC) analysis. Fifty-three images with 53 HCC nodules were evaluated. Tumor size ranged from 5 to 90 mm (mean 22.8 mm). Sensitivities and specificities for all techniques were calculated. RESULTS: ROC analysis showed the diagnostic ability significantly better with combined CTAP and CTHA (mean area under the ROC curve (Az)=0.95), or CTHA alone (Az=0.93) than CTAP alone (Az=0.87) (p<0.01). Combined CTAP and CTHA showed the best sensitivity (95.0%), followed by CTHA alone (88.1%) and CTAP alone (85.5%). The specificities of all three imaging techniques were relatively low (54.1% for combined CTAP and CTHA, 71.1% for CTHA alone, and 54.1% for CTAP alone) because of perfusion abnormalities of the liver parenchyma. CONCLUSION: The combination of CTAP and CTHA is superior to CTAP alone for detection of hypervascular HCCs. However, its specificity was relatively low in chronic liver damage. PMID- 11016768 TI - Contrast-enhanced color Doppler US in malignant portal vein thrombosis. AB - PURPOSE: To assess the role of contrast-enhanced color Doppler US in the differential diagnosis of benign and malignant portal vein thrombosis. MATERIAL AND METHODS: Fifty-six patients with portal vein thrombosis underwent color and power Doppler US examination before and after i.v. injection of galactose palmitic acid suspension. The criterion for diagnosing the presence and extension of thrombosis was the lack of visualization of portal vein flow in a segment of the portal vein. The criterion for diagnosing malignant portal vein thrombosis was the detection of pulsatile arterial flow, either hepatopetal or hepatofugal, in the thrombus. The US data were correlated with the types of portal vein thrombosis. RESULTS: Among the 56 patients, there were 40 benign and 16 malignant portal thromboses. Unenhanced Doppler US detected continuous blood flow in 24 benign thromboses and pulsatile in 3 malignant thromboses. No flow, either continuous or pulsatile, was detected in 16 cases with benign thrombosis and in 13 cases with malignant thrombosis. Contrast-enhanced Doppler US allowed to assess pulsatile flow in 15 cases with malignant thrombosis (94%). The use of pulsatile flow as diagnostic criterion of malignant thrombosis yielded a sensitivity of 57% and a specificity of 95% with conventional Doppler US, whereas contrast-enhanced US achieved a sensitivity of 94% and a specificity of 100%. CONCLUSION: Contrast-enhanced Doppler US is a reliable diagnostic tool for assessing malignant portal vein thrombosis. PMID- 11016769 TI - Transjugular intrahepatic portosystemic shunt. Experiences at a liver transplantation center. AB - OBJECTIVE: Transjugular intrahepatic portosystemic shunt (TIPS) placement is an established therapy for portal hypertension that leads to variceal bleeding or refractory ascites. We present experiences of the role of TIPS at a liver transplantation center. MATERIAL AND METHODS: One hundred and ten patients were referred to the Radiological Department for TIPS placement. One of the 110 patients had recurrent cirrhosis after liver transplantation with refractory ascites. Function of the TIPS was controlled with Doppler US at 1 day, 1 week, 4 weeks and 4 months after TIPS placement and subsequently every 3 months. Shunt insufficiency was supposed when the blood flow velocity within the stent tract was under 50 cm/s and was an indication for TIPS revision. RESULTS: TIPS was placed in 101 patients. After TIPS placement, 10 patients underwent liver transplantation. While waiting for the new liver, none of them developed variceal rebleeding, ascites or other complications of portal hypertension. Two of the 101 patients had episodes of rebleeding. The frequency of patients undergoing TIPS revision within the first year after TIPS placement was 67.5%, within the second year 38.0% and within the third year 24.4%. The revisions led to sufficient reduction of the portosystemic pressure gradient. CONCLUSION: In some liver transplant candidates, TIPS can be useful in minimizing the risk of complications of portal hypertension during the waiting time for a liver transplantation. TIPS can be monitored by Doppler US and revised if occlusion occurs. PMID- 11016770 TI - Miliary hepatic tuberculosis not associated with splenic or lung involvement. A case report. AB - Miliary hepatic involvement is a frequent finding on autopsy in patients with disseminated tuberculosis. Imaging studies may reveal hepatosplenomegaly and/or parenchymal inhomogeneity and, in a minority of cases, focal lesions, invariably associated with miliary lung disease. An unusual case of disseminated tuberculosis with manifestations of miliary hepatic involvement, abdominal and neck lymphadenopathy on US and CT without any evidence of active disease in the lungs, spleen or other organ, is described. PMID- 11016771 TI - Barium sulfate as contrast medium for evaluation of postoperative anastomotic leaks. AB - OBJECTIVE: To assess the value of barium swallow as a method for immediate re examination after the failure of an aqueous iodinated agent in detection of clinically suspected postoperative esophageal leakage, and as the initial study in asymptomatic postoperative patients. MATERIAL AND METHODS: Prospective examinations were performed in 114 postoperative patients, 95 males and 19 females (mean age 60 years). Initial esophagography with an aqueous contrast agent was performed in all cases. The patients who had no evidence of leakage underwent immediate re-examination with barium sulfate. Leakage volume was calculated in cm3. Clinical presentations of leakage before the examinations and the complications after the study were recorded. RESULTS: Leakage was shown with the initial study, using aqueous medium, in 23 patients (20%). The volume of leakage was between 0.05 and 36 cm3 (mean 3.95 cm3). Clinical signs and symptoms presented in 13 cases (56%). Fourteen of 91 patients (15%), who had negative results with the initial study, had evidence of leakage at barium swallow. The leakage volume ranged between 0.06 and 0.53 cm3 (mean 0.18 cm3). Clinical evidence of leakage was shown in 3/14 (21%) cases. No complications were detected over a 6-month period following the study. CONCLUSION: A barium swallow should follow a negative study using an aqueous agent in clinically suspected postoperative leakage, and could be considered as a safe initial study in postoperative patients where a leak is not suspected on clinical grounds. PMID- 11016772 TI - Ultrasonography in acute appendicitis. Body mass index as selection factor for US examination. AB - PURPOSE: Acute appendicitis is often difficult to diagnose and a negative laparotomy rate of about 25% is common. At Danderyd Hospital we started routine US in these patients, the aim being to estimate the sensitivity and the specificity for US when compared with the body mass index (BMI) of the patient. MATERIAL AND METHODS: All patient records were examined: During a period of 6 months 142 patients over 14 years of age were investigated with US. Their height and weight were noted and the BMI was calculated. RESULTS: The sensitivity for US examination was 0.76 in patients with a BMI < 25 but only 0.37 in patients with BMI > or = 25. This difference was statistically significant. CONCLUSION: US is a good method for examination of patients with BMI less than 25 but not in patients with BMI over 25. PMID- 11016773 TI - Ultrasonography and CT findings of a dermoid cyst of the cecum. A case report. AB - We report the US and CT findings of a dermoid cyst of the cecum, which is a very uncommon location for this entity. Pathological correlation and differential diagnosis are briefly discussed. PMID- 11016774 TI - Analysis of patellar cartilage. Comparison of conventional MR imaging and MR and CT arthrography in cadavers. AB - OBJECTIVE: To compare the sensitivity of conventional MR sequences, MR arthrography, and CT arthrography for the detection of cartilage lesions of the patella in cadavers. MATERIAL AND METHODS: Cartilage lesions in 10 cadaveric specimens were evaluated by MR imaging, including T1-weighted, proton density weighted and T2-weighted sequences, and fat-suppressed spoiled gradient recalled acquisition in the steady state (SPGR), MR arthrography including T1-weighted and SPGR sequences, and double-contrast CT arthrography including conventional and subtracted images. The sensitivities with regard to detection of lesions were compared to results from morphologic and histologic investigations of sectioned specimens. RESULTS: Twenty-one lesions were detected morphologically. For the detection of these lesions, sensitivities were as follows: T1-weighted images 33.3%; proton density-weighted images 85.7%; T2-weighted images 85.7%; SPGR images 80.9%; MR arthrography with T1-SE sequences 57.1%; MR arthrography with SPGR sequence 90.5%; and CT arthrography, both regular and subtracted images 85.7%. CONCLUSION: For noninvasive techniques, T2-weighted images revealed the highest sensitivity for the detection of patellar cartilage lesions, which was surpassed only by MR arthrography using the SPGR sequence. CT arthrography delineated surface irregularities but failed to demonstrate intrachondral lesions. PMID- 11016775 TI - Swedish metacarpophalangeal standards compared with previously published norms. AB - PURPOSE: Metacarpophalangeal lengths are used to create pattern profiles that are useful for assessment of skeletal dysplasias and malformation syndromes. The purpose of this study was to establish Swedish standards and compare them with previously published norms. MATERIAL AND METHODS: A group of healthy Swedish children was followed longitudinally from the age of 1 month to 18 years. The length of the tubular bones of the hand was measured on radiographs. In addition to the conventional measurements including epiphyses, diaphyseal lengths alone were recorded. RESULTS: Means and standard deviations of the metacarpophalangeal lengths are presented by gender and age. CONCLUSION: In spite of reasonably good correlation to previously published norms, demonstrated deviations encourage us to recommend the use of local standards whenever available. PMID- 11016776 TI - Nephrotoxicity of high- and low-osmolar contrast media. The protective role of amlodipine in a rat model. AB - PURPOSE: To evaluate the nephrotoxicity of high- and low-osmolar contrast media (HOCM, LOCM) on kidneys in Sprague-Dawley rats. The protective role of amlodipine was studied. MATERIAL AND METHODS: Forty rats of both sexes were randomly divided into 5 groups (n=8/group) and glycerine for inducing renal failure was given to all rats except controls. RESULTS: In diatrizoate-injected rats, blood urea nitrogen (BUN) and serum creatinine (SCr) were increased; levels of phospholipase A2 (PLA2), lipid peroxide (LPO) and calcium were also increased in renal tissues. There was no significant difference between LOCM (iohexol) animals and glycerol controls either in the renal levels of PLA2, LPO and calcium or in the levels of BUN and SCr. The histologic changes were milder in the LOCM animals than in the HOCM animals. In the group pretreated with amlodipine, no increase in the levels of BUN or SCr was discovered and the renal content of PLA2, LPO and calcium were significantly lower than in the HOCM group; the renal injuries induced by diatrizoate were alleviated. CONCLUSION: The HOCM, diatrizoate, was more toxic to rat kidneys than the LOCM iohexol; PLA2, LPO and calcium load played a role in producing renal function impairment induced by diatrizoate meglumine; amlodipine protected the renal tissue from nephrotoxicity induced by diatrizoate. PMID- 11016777 TI - Low- versus high-osmolality contrast media. AB - PURPOSE: To evaluate the effects of contrast media pharmokinetic differences on aortic enhancement at abdominal CT angiography and to determine whether these effects are of clinical relevance. MATERIAL AND METHODS: Two hundred and twelve patients referred for abdominal CT angiography were included in the study. All abdominal CT angiograms were performed with the same parameters (collimation 3 mm, pitch ratio 1.7, scan delay 30 s) after i.v. injection of 120 ml of contrast medium at 3 ml/s. After randomization, patients received either iobitridol 300 (low-osmolar, 300 mg I/ml), iobitridol 350 (low-osmolar, 350 mg I/ml) or ioxithalamate 350 (high-osmolar, 350 mg I/ml). The time attenuation curves obtained with the three contrast media were compared. RESULTS: The time attenuation curve obtained with ioxithalamate 350 was not parallel to those obtained with iobitridol 300 and iobitridol 350. Mean peak enhancements obtained with iobitridol 350 and ioxithalamate 350 were not significantly different but iobitridol 350 provided higher mean peak enhancement than iobitridol 300. Mean delays of the peak enhancements were the same with the three contrast media. After peak enhancement, the decrease of aortic opacification under a selected threshold of 200 HU was significantly slower with iobitridol 350 than with iobitridol 300 and ioxithalamate 350, whereas iobitridol 300 and ioxithalamate 350 showed no significant differences. CONCLUSION: For a given iodine concentration, low-osmolality contrast media provide longer aortic opacification and may be recommended for CT angiography when long acquisition times are needed. PMID- 11016778 TI - Ion channels and receptors: molecular targets for behavioral evolution. AB - Ion channels and receptors play critical roles in shaping neuronal activity, and thus are appropriate targets for evolutionary change to generate new behaviors. In this review, the evolution and differentiation of the many voltage-gated ion channels and transmitter-activated receptors is summarized; these channels and receptors evolved very early, and with some exceptions all species with nervous systems use similar sets of channels and receptors. Several examples are given of mechanisms for species-specific behavioral evolution that arise from mutations involving the structure, alternative splicing, level of expression, targeting and modulation of these important neural proteins. PMID- 11016779 TI - Aminergic neuron systems of lobsters: morphology and electrophysiology of octopamine-containing neurosecretory cells. AB - In the American lobster (Homarus americanus) the biogenic amines serotonin and octopamine appear to play important and opposite roles in the regulation of aggressive behavior, in the establishment and/or maintenance of dominant and subordinate behavioral states and in the modulation of the associated postural stances and escape responses. The octopamine-containing neurosecretory neurons in the thoracic regions of the lobster ventral nerve cord fall into two morphological subgroups, the root octopamine cells, a classical neurohemal group with release regions along second thoracic roots, and the claw octopamine cells, a group that selectively innervates the claws. Cells of both subgroups have additional sets of endings within neuropil regions of ganglia of the ventral nerve cord. Octopamine neurosecretory neurons generally are silent, but when spontaneously active or when activated, they show large overshooting action potentials with prominent after-hyperpolarizations. Autoinhibition after high frequency firing, which is also seen in other crustacean neurosecretory cells, is readily apparent in these cells. The cells show no spontaneous synaptic activity, but appear to be excited by a unitary source. Stimulation of lateral or medial giant axons, which excite serotonergic cells yielded no response in octopaminergic neurosecretory cells and no evidence for direct interactions between pairs of octopamine neurons, or between the octopaminergic and the serotonergic sets of neurosecretory neurons was found. PMID- 11016780 TI - A central pattern generator underlies crawling in the medicinal leech. AB - Crawling in the medicinal leech has previously been thought to require sensory feedback because the intact behavior is strongly modulated by sensory feedback and because semi-intact preparations will only crawl if they can move freely. Here we show that an isolated leech nerve cord can produce a crawling motor pattern similar to the one seen in semi-intact preparations, which consists of an anterior-to-posterior wave of alternating excitatory circular and longitudinal motor neuron bursts in each segment. The isolated cord also reproduces the patterns of activity seen in semi-intact preparations for several other kinds of cells: the dorsal inhibitor cell 1, the ventral excitor cell 4, and the annulus erector motor neuron. Because this correspondence is so strong, there must be a central pattern generator in the isolated cord that can produce the basic motor pattern for crawling without sensory feedback. A quantitative analysis of the isolated motor pattern, however, reveals that isolated and semi-intact preparations have longer periods than the intact behavior and that there are deficiencies in the timing of motor neuron bursts in the isolated pattern. These results suggest that sensory feedback modulates the isolated central pattern generator to help produce the normal motor pattern. PMID- 11016781 TI - Spontaneous modulations of the electric organ discharge in the weakly electric fish, Apteronotus leptorhynchus: a biophysical and behavioral analysis. AB - Brown ghosts, Apteronotus leptorhynchus, are weakly electric gymnotiform fish whose wave-like electric organ discharges are distinguished by their enormous degree of regularity. Despite this constancy, two major types of transient electric organ discharge modulations occur: gradual frequency rises, which are characterized by a relatively fast increase in electric organ discharge frequency and a slow return to baseline frequency; and chirps, brief and complex frequency and amplitude modulations. Although in spontaneously generated gradual frequency rises both duration and amount of the frequency increase are highly variable, no distinct subtypes appear to exist. This contrasts with spontaneously generated chirps which could be divided into four "natural" subtypes based on duration, amount of frequency increase and amplitude reduction, and time-course of the frequency change. Under non-evoked conditions, gradual frequency rises and chirps occur rather rarely. External stimulation with an electrical sine wave mimicking the electric field of a neighboring fish leads to a dramatic increase in the rate of chirping not only during the 30 s of stimulation, but also in the period immediately following the stimulation. The rate of occurrence of gradual frequency rises is, however, unaffected by such a stimulation regime. PMID- 11016782 TI - GABAergic inhibition shapes frequency tuning and modifies response properties in the superior olivary nucleus of the leopard frog. AB - The role of gamma-aminobutyric acid (GABA)ergic inhibition in shaping the excitatory frequency tuning of 74 neurons in the superior olivary nucleus of the leopard frog, Rana pipiens, was studied using iontophoretic application of the GABA(A) receptor antagonist, bicuculline methiodide. For 37 neurons, bicuculline application broadened and/or changed the configuration of the excitatory frequency-tuning curve. Results indicate that GABA-mediated inhibition not only sharpens the tuning curves of neurons but also plays a critical role in creating new frequency tuning properties in the superior olivary nucleus. Bicuculline application affected other neuronal response properties as well. Spontaneous firing rate increased 11-338% for 18 of 59 neurons. For 32 of 58 neurons there was an increase in stimulus-evoked discharge rate and a change in rate-level function. There was no qualitative effect on the discharge pattern of 60 neurons, though 2 tonically responding neurons did show an increase (> 30%) in response duration. Additional roles for GABAergic inhibition in monaural signal analysis are discussed. PMID- 11016783 TI - Source levels and the estimated active space of bottlenose dolphin (Tursiops truncatus) whistles in the Moray Firth, Scotland. AB - This study measured SPLs of whistles of wild bottlenose dolphins (Tursiops truncatus) in the Moray Firth, Scotland, and estimated their active space, i.e. the distance at which another dolphin can perceive the whistle of a conspecific. Whistling dolphins were localized with a dispersed hydrophone array by comparing differences in the times of arrival of a whistle at different hydrophones. The mean source level for whistles was 158 +/- 0.6 dB re. 1 microPa. The maximum was 169 dB re. 1 microPa. The active space of these whistles was calculated taking into account transmission loss, ambient noise, the critical ratios and the auditory sensitivity of this species. The estimated radius of the active space of unmodulated whistles between 3.5 kHz and 10 kHz produced at maximum source level ranged from 20 km to 25 km in a habitat of 10 m depth and at sea state 0. At sea state 4 it ranged from 14 km to 22 km. For whistles of 12 kHz it dropped to 1.5-4 km. The results suggest that whistles can be used to maintain group cohesion over large distances but also that dolphins that researchers consider to belong to separate groups might be in acoustic contact. PMID- 11016784 TI - Visual pigments, cone oil droplets and ocular media in four species of estrildid finch. AB - A microspectrophotometric study was conducted on the retinal photoreceptors of four species of bird: cut-throat finches (Amadina fasciata), gouldian finches (Erythrura gouldiae), white-headed munias (Lonchura maja) and plum-headed finches (Neochmia modesta). Spectral characteristics of the photoreceptors in all four species were very similar. Rods contained a medium-wavelength-sensitive visual pigment with a wavelength of maximum absorbance at 502-504 nm. Four spectrally distinct types of single cone contained a visual pigment with wavelength of maximum absorbance at either 370-373 nm (ultraviolet-sensitive), 440-447 nm (short-wavelength-sensitive); 500 nm (medium-wavelength-sensitive) or 562-565 nm (long-wavelength-sensitive). Oil droplets in the ultraviolet-sensitive single cones showed no detectable absorption between 330 nm and 800 nm. Oil droplets in the short-, medium-, and long-wavelength-sensitive single cones had cut-off wavelengths at 415-423 nm, 510-520 nm and 567-575 nm, respectively. Double cones contained the visual pigment with wavelength of maximum absorbance at 562-565 nm observed in long-wavelength-sensitive single cones. Only the principal member of the double cone pair contained an oil droplet (P-type, cut-off wavelength at 414 489 nm depending on species and retinal location). Spectral transmittance of the intact ocular media of each species was measured along the optic axis. Wavelengths of 0.5 transmittance for all species were very similar (316-318 nm). PMID- 11016785 TI - Directionality in the mechanical response to substrate vibration in a treehopper (Hemiptera: Membracidae: Umbonia crassicornis). AB - The use of substrate vibrations in communication and predator-prey interactions is widespread in arthropods. In many contexts, localization of the vibration source plays an important role. For small species on solid substrates, time and amplitude differences between receptors in different legs may be extremely small, and the mechanisms of vibration localization are unclear. Here we ask whether directional information is contained in the mechanical response of an insect's body to substrate vibration. Our study species was a membracid treehopper (Umbonia crassicornis) that communicates using bending waves in plant stems. We used a bending-wave simulator that allows precise control of the frequency, intensity and direction of the vibrational stimulus. With laser-Doppler vibrometry, we measured points on the substrate and on the insect's thorax and middle leg. Transfer functions showing the response of the body relative to the substrate revealed resonance at lower frequencies and attenuation at higher frequencies. There were two modes of vibration along the body's long axis, a translational and a rotational mode. Furthermore, the transfer functions measured on the body differed substantially depending on whether the stimulus originated in front of or behind the insect. Directional information is thus available in the mechanical response of the body of these insects to substrate vibration. These results suggest a vibration localization mechanism that could function at very small spatial scales. PMID- 11016786 TI - Can the circadian system of a diurnal and a nocturnal rodent entrain to ultraviolet light? AB - Spectral measurements of sunlight throughout the day show close correspondence between the timing of above ground activity of the European ground squirrel and the presence of ultraviolet light in the solar spectrum. However, in a standard entrainment experiment ground squirrels show no entrainment to ultraviolet light, while Syrian hamsters do entrain under the same protocol. Presented transmittance spectra for lenses, corneas, and vitreous bodies may explain the different results of the entrainment experiment. We found ultraviolet light transmittance in the colourless hamster lens (50% cut-off at 341 nm), but not in the yellow ground squirrel lens (50% cut-off around 493 nm). Ultraviolet sensitivity in the ground squirrels based upon possible fluorescence mechanisms was not evident. Possible functions of ultraviolet lens filters in diurnal mammals are discussed, and compared with nocturnal mammals and diurnal birds. Species of the latter two groups lack ultraviolet filtering properties of their lenses and their circadian system is known to respond to ultraviolet light, a feature that does not necessarily has to depend on ultraviolet photoreceptors. Although the circadian system of several species responds to ultraviolet light, we argue that the role of ultraviolet light as a natural Zeitgeber is probably limited. PMID- 11016787 TI - Integration of temperature and olfactory information in cockroach antennal lobe glomeruli. AB - Individual neurons in the antennal lobe of the cockroach not only respond to warming, cooling and the odor of lemon oil but they also integrate the responses to simultaneously occurring temperature and olfactory stimuli. This integration results in an increase or decrease of the neuron's activity as compared to its responses to the temperature stimuli presented alone. The mean gain for a change in temperature in the warm and cold direction is 9.5 (imp s(-1)) degrees C(-1) and 10.2 (imp s(-1)) degrees C(-1), respectively. Thus, the average neuron elevates its impulse frequency by 1 imp s(-1) when temperature is increased by 0.1 degree C or decreased by 0.09 degree C. Examination of response scatter reveals that the difference required between two warm or two cold stimuli to be discriminated is 0.5 degree C. Similar values for gain and resolving power are obtained for the enhanced responses to the warm-odor and the cold-odor stimulus combinations. The neurons described are: (1) local interneurons innervating a number of glomeruli distributed within the antennal lobe, and (2) projection neurons collecting information from single glomeruli at 140-280 microm from the surface of the antennal lobe and providing links with the calyces of the mushroom bodies and the lateral lobe of the protocerebrum. PMID- 11016788 TI - Coupling of visual to auditory cues during phonotactic approach in the phaneropterine bushcricket Poecilimon affinis. AB - In the duetting bushcricket species Poecilimon affinis the male calls at intervals of several seconds and is guided to the female by its short response clicks, which release phonotaxis only when perceived by the male during its sensory time window (40-170 ms after his call). The accuracy of phonotaxis in this acoustically open-loop system was investigated on a locomotion compensator with and without optical cues available. Phonotaxis in darkness was strongly meandrous with numerous roundabouts, while in a structured surrounding the oscillating course was attenuated. With a landmark available the male was able to maintain a straight course to the female. This is achieved by coupling of visual cues to an acoustically detected direction. Thus, in this species, the acoustic cues, which in the songs of continuously singing crickets and bushcrickets are permanently present, are replaced by optical ones. Restricting localization of female clicks to a short time window and using optical cues for target tracking allows straight orientation, even when guided by very short signals at long repetition intervals. PMID- 11016789 TI - Arrangement of optical axes and spatial resolution in the compound eye of the female blowfly Calliphora. AB - We determined the optical axes of ommatidia in the wild-type female blowfly Calliphora by inspecting the deep pseudopupil in large parts of the compound eye. The resulting map of optical axes allowed us to evaluate the spatial resolution in different parts of the eye in terms of interommatidial angles as well as the density of optical axes, and to estimate the orientation of ommatidial rows along the hexagonal eye lattice. The optical axes are not homogeneously distributed over the eye. In the frontal visual field the spatial resolution is about two times higher than in its lateral part and about three times higher as compared to the eye's dorsal pole region. The orientation of the ommatidial rows along the eye lattice is not the same for different regions of the eye but changes in a characteristic way. The inter-individual variability in the orientation of the ommatidial rows is estimated to be smaller than 8 degrees . The characteristic arrangement of the ommatidial lattice is discussed as an adaptation for efficient evaluation of optic flow as induced during self-motions of the animal. PMID- 11016790 TI - Involvement of a midbrain vocal nucleus in the production of both the acoustic and postural components of crowing behavior in Japanese quail. AB - Many bird species produce vocalizations which are accompanied by distinctive postural displays, but the neural mechanisms that allow such integrated production of vocal and postural motor patterns are not understood. In the crowing behavior of Japanese quail, a characteristic vocal pattern is accompanied by and coordinated with a postural display that consists of a sequence of rapid, patterned head movements. The purpose of the present study was to investigate the role of a midbrain vocal nucleus, the nucleus intercollicularis, in the production of the acoustic and postural components of crowing in quail. Brief electrical stimuli were applied to the nucleus intercollicularis during spontaneously emitted crows in quail with chronically implanted electrodes, to determine if perturbing neural activity in the nucleus intercollicularis resulted in a disruption of ongoing crowing behavior. The most common effect of such stimuli was a concurrent, premature termination of both the acoustic and head movement components of the crow. These results imply that the nucleus intercollicularis plays a role in the production of both the acoustic and postural components of crowing in quail. PMID- 11016791 TI - Acute and chronic effects of an aromatase inhibitor on territorial aggression in breeding and nonbreeding male song sparrows. AB - Many studies have demonstrated that male aggression is regulated by testosterone. The conversion of testosterone to estradiol by brain aromatase is also known to regulate male aggression in the breeding season. Male song sparrows (Melospiza melodia morphna) are territorial not only in the breeding season, but also in the nonbreeding season, when plasma testosterone and estradiol levels are basal. Castration has no effect on nonbreeding aggression. In contrast, chronic (10 day) aromatase inhibitor (fadrozole) treatment decreases nonbreeding aggression, indicating a role for estrogens. Here, we show that acute (1 day) fadrozole treatment decreases nonbreeding territoriality, suggesting relatively rapid estrogen effects. In spring, fadrozole decreases brain aromatase activity, but acute and chronic fadrozole treatments do not significantly decrease aggression, although trends for some behaviors approach significance. In gonadally intact birds, fadrozole may be less effective at reducing aggression in the spring. This might occur because fadrozole causes a large increase in plasma testosterone in intact breeding males. Alternatively, estradiol may be more important for territoriality in winter than spring. We hypothesize that sex steroids regulate male aggression in spring and winter, but the endocrine mechanisms vary seasonally. PMID- 11016792 TI - Neuronal pathways from low-threshold muscle and cutaneous afferents innervating tail to trunk muscle motoneurons in the cat. AB - We studied neuronal pathways from low-threshold muscle (group I, II) and cutaneous afferents (group A(alpha)beta) innervating the tail to motoneurons innervating trunk muscles (m. iliocostalis lumborum and m. obliquus externus abdominus) in 18 spinalized cats. Stimulation of group I muscle afferents produced excitatory postsynaptic potentials or excitatory postsynaptic potentials followed by inhibitory postsynaptic potentials in all motoneurons innervating the m. iliocostalis lumborum which showed effects (32%), and predominantly inhibitory postsynaptic potentials in motoneurons innervating the m. obliquus externus abdominus (47%). Stimulation of group I+II afferents produced significant increases of the incidence of motoneurons showing postsynaptic potentials (the notoneurons innervating the m. iliocostalis lumborum, 87%; the motoneurons innervating the m. obliquus externus abdominus, 82%). The effects of low threshold cutaneous afferents were bilateral, predominantly producing inhibitory postsynaptic potentials in motoneurons innervating both muscles. These results suggest that neuronal pathways from muscle afferents to back muscle motoneurons mainly increase the stiffness of the trunk to maintain its stability, while those to abdominal muscles help to extend the dorsal column by decreasing their activities. The results also indicate that neuronal pathways from cutaneous afferents to trunk motoneurons functionallY disconnect the tail from the trunk. PMID- 11016793 TI - Angiotensin-converting enzyme gene insertion/deletion (I/D) polymorphism in hypertensive patients with different degrees of obstructive sleep apnea. AB - To investigate the role of the angiotensin-converting enzyme gene (ACE) insertion (I)/deletion (D) polymorphism in hypertensive patients with different degrees of obstructive sleep apnea (OSA). A case-control study was performed. One hundred seventy four Chinese subjects were divided into four groups depending on the severity of OSA as follows: 1) normal control group (NC, n=68), 2) isolated hypertension group (HT, n=45), 3) hypertensive patients with mild OSA group (MO, n=27), and 4) hypertensive patients with moderate to severe OSA group (MSO, n=34). The distribution of ACE gene I/D allele and genotypes were analyzed in the subject population, as was an OSA pedigree. The study showed that the frequency of ACE gene I/D polymorphism differed significantly among the four groups. The frequency of I allele and II genotype were significantly higher in the MSO group than in the other groups (p<0.05). The distribution of I allele and II genotype showed no significant difference between any of the other groups (p>0.05, respectively). Meanwhile the higher frequency of I allele and II genotype was observed in the OSA pedigree. The higher frequency of ACE gene I allele and II genotype were closely associated with the hypertensive patients with MSO. The inherited factors played an important role in the pathogenesis of hypertensive patients with MSO. PMID- 11016794 TI - Comparative studies of diet-related factors and blood pressure among Chinese and Japanese: results from the China-Japan Cooperative Research of the WHO-CARDIAC Study. Cardiovascular Disease and Alimentary Comparison. AB - We aimed to compare the differences in diet-related factors and their associations with blood pressure (BP) between Chinese and Japanese. A total of 1,151 Chinese (M/F: 551/600) and 1,681 Japanese (782/899), aged 48-56 years, were studied using a multi-center cross-sectional study design. This work was a constituent part of the World Health Organization (WHO) Cardiovascular Disease and Alimentary Comparison (CARDIAC) Study. Measurements included in the present report were BP, body mass index (BMI), serum total cholesterol (TC), 24-h urinary sodium, potassium, calcium, magnesium, creatinine, 3-Methylhistidine (3MH, a marker of animal protein intake) and taurine (a marker of seafood intake) excretion levels. Results were as follows: (a) Japanese men had a significantly higher prevalence of hypertension than the Chinese (34.4% vs. 20.5%, p<0.01). After adjustment for age, Japanese men had a significantly higher mean systolic and diastolic BP (SBP, DBP), and Japanese women had a significantly higher DBP than the Chinese subjects overall (p<0.01, respectively). (b) Japanese had significantly higher mean BMI, TC and sodium excretion, and lower mean magnesium excretion than Chinese (p<0.01). (c) In the Japanese sample, multiple linear regression analyses (using a stepwise procedure) showed that SBP had a significant positive association with BMI and sodium excretion, and a significant negative association with magnesium excretion, while DBP had a significant positive association with BMI and a significant negative association with the 3MH to creatinine ratio (3MH/Cre). In the Chinese sample, both SBP and DBP showed a significant positive association with BMI and sodium, and a significant negative association with 3MH/Cre. In conclusion, Japanese had significantly higher mean BP than Chinese. The differences in BP may have been partly attributable to differences in various diet-related factors, particularly in BMI, sodium, magnesium-rich foods and animal protein intake, between the two populations. PMID- 11016795 TI - Effects of body weight control on changes in blood pressure: three-year follow-up study in young Japanese individuals. AB - Numerous epidemiological studies have shown a close relationship between obesity and hypertension. However, there have been few reports on the relationship between changes in the body weight and blood pressure of lean to normal-weight young subjects. The purpose of this study was to investigate the effects of body weight control on blood pressure in lean to obese young Japanese individuals in a 3-year follow-up study. University students (3,558 males and 1,418 females, aged 18.6+/-0.8 in 1994) were classified into 4 groups according to the baseline body mass index (BMI), and were followed up for 3 years. Among male students, changes in body weight were significantly correlated with changes in blood pressure during the 3 years in all 4 BMI groups, and the correlation coefficient was larger in the group with higher baseline BMI. Positive correlations between changes in body weight and changes in heart rate were noted only in the obese and mildly-obese groups. Also in female students, positive correlations were observed between changes in body weight and changes in blood pressure in lean to obese groups. However, no correlations between changes in body weight and changes in heart rate were noted in any of the female groups. To summarize, close correlations were observed between changes in body weight and those in blood pressure during the 3 years in both male and female university students. These findings suggest the importance of body weight control not only in obese but also in normal to mildly-obese young subjects in reducing or preventing an increase in blood pressure. There could be, however, a gender difference in the effects of body weight change on heart rate. PMID- 11016796 TI - Circulating adrenomedullin in erythrocopietin-induced hypertension. AB - Levels of adrenomedullin (AM) have been shown to be elevated in hypertension and chronic renal failure, suggesting that AM plays a role in the pathogenesis of these diseases. The objective of the present study was to investigate whether circulating AM is involved in erythropoietin (Epo)-induced hypertension in patients with renal anemia due to progressive renal disease. Following treatment with 6,000 IU of Epo once a week, the hematocrit (Ht) rose significantly from 25.9+/-4.0 to 33.4+/-3.3% (n=54, p<0.001) with an overall rate of increase in Ht of 0.43+/-0.04%/week. In response to treatment with Epo, a rise in mean blood pressure of >10 mmHg (Epo-induced hypertension) was found in 22% (12/54 cases) of the patients enrolled. There was no difference in the rate of Ht increase between patients with and without Epo-induced hypertension. There was a significant positive correlation between mature AM and serum creatinine (Cr) concentration before treatment with Epo. However, no correlation was found between the plasma concentration of total AM and serum Cr concentration. Long-term treatment with Epo did not influence plasma concentration of either mature AM or total AM in patients developing hypertension during the study period. These results suggest that circulating AM may play a role in the progression of renal disease. However, the present study does not support the notion that circulating AM is associated with the pathogenesis of Epo-induced hypertension. It is too early yet to claim that there is no AM-mediated mechanism in Epo-induced hypertension. PMID- 11016797 TI - Insulin sensitivity and calcium homeostasis in young, lean, normotensive male subjects. AB - Insulin resistance is known to be closely related to essential hypertension. It has been hypothesized that abnormal calcium homeostasis both at the cellular level and in the whole body plays a substantial role in hypertension associated with insulin resistance. We attempted to determine the relationships among insulin sensitivity, blood pressure (BP), body mass index (BMI), and calcium related parameters in young, lean, normotensive male subjects with extreme susceptibilities to hypertension, and to investigate the effects of euglycemic hyperinsulinemia on calcium-related parameters. Seven young, lean, normotensive male subjects with family histories of essential hypertension and 10 age-matched controls without any parental cardiovascular events were enrolled. Insulin sensitivity measurement by the euglycemic hyperinsulinemic clamp technique and a 75-g oral glucose tolerance test were performed. Calcium-related parameters, including intracellular Ca2+ levels in platelets, were measured simultaneously. Diastolic BP was inversely correlated with insulin sensitivity in vivo (M-value). Insulin sensitivity was inversely correlated with BMI and with intracellular Ca2+ in platelets. In the multivariate stepwise regression analysis using both diastolic BP and insulin sensitivity as dependent variables, BMI was found to be a determinant independent variable. Euglycemic hyperinsulinemia decreased intact parathyroid hormone levels and increased fractional excretion of calcium. In conclusion, BMI rather than a family history of hypertension plays a determinant role on the regulation of diastolic BP and insulin sensitivity even in young, lean, normotensive male subjects with extreme predispositions for the development of hypertension. Hyperinsulinemia decreased intact parathyroid hormone levels and increased fractional excretion of calcium. PMID- 11016798 TI - Control of blood pressure and lifestyle-related risk factors in elderly Japanese hypertensive subjects. AB - The aim of this study was to analyze the treatment of elderly hypertensive patients by Japanese physicians specializing in hypertension. We enrolled 939 patients with hypertension who were treated in the outpatient clinics of 11 hospitals in 1995; 793 of these patients (388 men and 405 women; mean age, 66.6+/ 9.0 years) received follow-up examinations in 1996, and the data on these patients was used for the present analysis. Blood pressure (BP), body mass index, lifestyle, and laboratory data were analyzed in all patients. The average BP was 143+/-16/81+/-10 mmHg in 1995 and 142+/-15/80+/-10 mmHg in 1996. The patients whose baseline BP was at the level of Grade 2 or 3 in the WHO-ISH classification (n=117) were characterized by a higher women-to-men ratio, higher age, a higher serum total cholesterol concentration, and higher QRS voltage. In these patients, from 1995 to 1996, the average BP significantly decreased, whereas fasting plasma glucose, serum total cholesterol and serum creatinine concentrations showed only negligible changes. In 220 patients (28%), BP was <140/<90 mmHg at both the initial and the follow-up examinations, whereas 351 patients (44%) were hypertensive in both 1995 and 1996. Thirty-three percent of the patients were smokers. More smokers than nonsmokers had had prior cardiovascular events, diabetes mellitus, or overt proteinuria. In conclusion, the average BP level among the patients treated by Japanese physicians specializing in hypertension was somewhat higher than that recommended by WHO-ISH Guidelines (1999). Patient education to control lifestyle-related risk factors, particularly to stop smoking, should be emphasized. PMID- 11016799 TI - The influence of the peripheral reflection wave on left ventricular hypertrophy in patients with essential hypertension. AB - The objective of this study was to clarify the relationship between afterload, which consists mainly of the vascular reflection wave, and left ventricular hypertrophy in patients with untreated essential hypertension using the fingertip photoplethysmogram (PTG) and second derivative wave (SDPTG) methods, the simplest and most convenient tools for pulse wave analysis. The augmentation index (AI) is defined as the ratio of the height of the late systolic peak, augmented by the peripheral reflection wave, to that of the early systolic peak caused mainly by left ventricular ejection in the pulse. Increased AI of the PTG and negative d/a, obtained by multiplying the ratio of the late re-decreasing wave (d wave) to the initial positive wave (a wave) of the SDPTG by -1, have the same meaning as increased ascending aortic AI. The left brachial artery blood pressure was measured in 60 patients. The PTG and SDPTG of the right second finger were recorded by a digital photoplethysmograph. The left ventricular mass index (LVMI) was investigated by ultrasonography. Subjects were assigned to one of two groups: a low AI (AI of PTG<1.6; group 1) or a high AI (AI of PTG> or =1.6; group 2) group. LVMI was significantly higher in group 2 than in group 1. In the study group as a whole, the LVMI was positively correlated with both the AI of PTG (r=0.60, p<0.0001) and negative d/a (r=0.63, p<0.0001). An increase in the LVMI was seen in subjects with an augmented late systolic component in the waveform. It was concluded that an increase in the peripheral reflection wave on the left ventricle is one of the important factors causing cardiac hypertrophy in patients with hypertension. PMID- 11016800 TI - Urinary excretions of albumin and type IV collagen in normotensive and hypertensive subjects. AB - Plasma albumin leaks into urine as a result of glomerular hypertension and basement membrane injury, while urinary type IV collagen derives from mesangial matrix and glomerular basement membrane. The purpose of this study was to elucidate the pathophysiological significance of these urinary microproteins as an indicator of cardiovascular organ injuries in hypertension. In health-checkup participants without diabetes, proteinuria, or microhematuria, and who were not being treated for hypertension or any other disease at the time of enrollment, urinary albumin and type IV collagen were measured and their relations to organ injuries and cardiovascular risk factors were evaluated. Of 1,079 subjects (40- to 65-year-old; 256 men and 823 women) enrolled in the study, 120 (11.1%) had untreated hypertension exceeding 140/90 mmHg. Urinary albumin was positively correlated with both age (r=0.16, p<0.001) and systolic blood pressure (r=0.27, p<0.001). Urinary type IV collagen was not only positively correlated with age (r=0.12, p<0.001) and diastolic blood pressure (r=0.14, p<0.001) but also negatively correlated with blood hemoglobin (r=-0.12, p<0.001). Urinary albumin, but not type IV collagen, had a significant relation to electrocardiographic signs of left ventricular hypertrophy (p=0.012) and retinal arteriosclerosis on fundoscopy (p <0.001). Thus both albumin and type IV collagen would seem to have increased in association with age and hypertension in this cohort. It is suggested that urinary albumin is an indicator not only of renal injury, but also possibly of development of cardiac hypertrophy and arteriosclerotic changes. Urinary type IV collagen, on the other hand, may be associated with renal tissue injuries that affect erythrokinetics. PMID- 11016801 TI - Effect of celiprolol on cardiac hypertrophy in hypertension. AB - The present study was undertaken to clarify whether celiprolol and atenolol, beta1-selective beta blockers with and without intrinsic sympathomimetic activity (ISA), respectively, might improve ischemic damage in the isolated perfused hearts of spontaneously hypertensive rats (SHR), and whether long-term treatment with celiprolol may reduce left ventricular hypertrophy (LVH) in patients with essential hypertension. Atenolol (50 mg/kg/day) or celiprolol (300 mg/kg/day) for 7 weeks significantly reduced the blood pressure in SHR to the same degree, and both drugs decreased the heart rate, but the magnitude of the fall in heart rate was significantly higher with atenolol treatment than with celiprolol treatment. Both treatments significantly reduced the ratio of LV weight to body weight in SHR and significantly improved the coronary reserve in SHR to the same extent. Both treatments significantly improved the extent of recovery of the pressure rate product and the extent of percent recovery of the coronary flow after reperfusion following 30 min of ischemia in SHR. Celiprolol treatment in patients with essential hypertension for 12 months significantly decreased interventricular septal thickness (IVST)+LV posterior wall thickness (PWT) and LV mass index (LVMI), but there was no significant correlation between IVST+PWT or LVMI and blood pressure before and after treatment. IVST+PWT and LVMI were significantly decreased after 3 months of treatment and these LVH indices were significantly smaller after 6 and 12 months of treatment than after 3 months of treatment. In conclusion, both celiprolol and atenolol treatment reduced LVH and improved the ischemic damage in SHR. In essential hypertensive patients with LVH, celiprolol treatment effectively reduced blood pressure and achieved LVH regression. PMID- 11016802 TI - A new murine model for atherosclerosis with inflammation in the periodontal tissue induced by immunization with heat shock protein 60. AB - It has recently become apparent that the anti-heat shock protein (HSP) antibody plays an important role in the pathogenesis of atherosclerosis. We studied whether immunization with human HSP60 could lead to atherosclerosis in mice. We attempted to induce atherosclerosis in C57BL/6NJcl mice by immunization with human HSP60 under a high-cholesterol diet. The size of fatty streak lesions was significantly enhanced in mice immunized with human HSP60 under a high cholesterol diet relative to the number in control mice receiving a high cholesterol diet alone. In addition, these new atherosclerotic model mice showed lesions of inflammation in the periodontal tissue. This new model may thus provide theoretical support for the clinical observation that patients suffering from periodontitis are frequently found to have atherosclerosis. The cytokine ratio of interferon-gamma/interleukin-4 in the high-cholesterol diet group was significantly higher than that in the standard chow group (p<0.05). This suggests the presence of a predominantly Th1-type immune response in atherosclerosis. PMID- 11016803 TI - Non-corresponding effects of an angiotensin-converting enzyme inhibitor on cardiac and vascular hypertrophy in spontaneously hypertensive rats. AB - Angiotensin-converting enzyme inhibitors (ACEIs) may have different effects on cardiac hypertrophy than on vascular hypertrophy. Arginine vasopressin (AVP) may promote cardiac hypertrophy. Our aims were (1) to simultaneously examine the chronic effects of ACEIs on hypertrophy of the heart and hypertrophy of the coronary and renal interlobular arteries, and (2) to clarify the relation between AVP concentration (AVPC) and cardiac hypertrophy. ACEI (delapril: 30 mg/kg/day) or vehicle (5% arabic gum) was administered in a preventive (4 to 28 weeks of age) or a therapeutic (12-24 weeks of age) protocol in spontaneously hypertensive rats. In both protocols, delapril produced a slight but significant decrease in systolic blood pressure. In the therapeutic protocol, the weight of the left ventricle (mean+/-SE) was lower (p<0.05) in the ACEI group (64+/-2 mg/100 g body weight) than in the control group (69+/-1 mg/100 g body weight). Plasma renin activity was significantly higher in the ACEI group than in the control group in both the preventive (p <0.01) and therapeutic (p<0.01) protocols. In the therapeutic protocol, AVPC was significantly (p<0.05) lower in the ACEI group than in the control group. AVPC was significantly (p=0.02, r=0.46) correlated with the weight of the left ventricle in the therapeutic protocol. For both protocols, no differences were noted between the ACEI and control groups in the vascular hypertrophy of the coronary and renal interlobular arteries. We conclude that (1) the preventive or therapeutic effect of ACEIs on hypertrophy may not be the same in the heart as in the coronary and renal arteries; and (2) AVP was significantly correlated with the left ventricular weight. This indicates that AVP could play a role in the etiology of cardiac hypertrophy in SHR. PMID- 11016804 TI - Possible involvement of endothelin-1 in cardioprotective effects of benidipine. AB - Benidipine hydrochloride has been developed as an antagonist for the L-type calcium channel and is used as an anti-hypertensive drug. But recent studies have reported that benidipine exerts not only antihypertensive actions but also anti hypertrophic actions on cardiac muscles. Endothelin-1 (ET-1), one of the endogenous pathological humoral factors of cardiovascular diseases such as hypertension and heart failure, has a strong vasoconstrictive action and could induce hypertension and cardiac hypertrophy. So, it is a matter of great interest whether or not calcium antagonists can decrease cardiac hypertrophy induced by the pathological vasoactive substances such as ET-1. Thus, the present study was designed to elucidate the effects of benidipine on cardiac hypertrophy, and particularly on the interaction with ET-1, using neonatal rat cardiac myocytes (MCs) and cardiac non-myocytes (NMCs) culture systems. Cells were cultured with or without ET-1, benidipine, and nifedipine and the effects of calcium antagonists on cardiac hypertrophy were evaluated by incorporations of [3H] leucine and [3H]-thymidine into MCs and/or NMCs. Benidipine significantly decreased the ET-1-induced increase of [3H]-leucine and [3H]-thymidine uptake into cardiac MCs and NMCs, whereas no significant effects of nifedipine were observed. Furthermore, benidipine (10(-8)M) attenuated ET-1 secretions from NMCs. In summary, benidipine at least partially decreased the cardiac hypertrophy induced by paracrine mechanisms through its attenuation of ET-1 secretions from NMCs. Benidipine could thus be a useful tool for preventing cardiac hypertrophy due to hypertension. PMID- 11016805 TI - Failure of probucol to prolong survival in salt-loaded stroke-prone spontaneously hypertensive rats. AB - We examined the effect of probucol, a lipid-lowering agent with strong antioxidant properties, on neurological events and survival in stroke-prone spontaneously hypertensive rats (SHRSP). Rapid onset of stroke was induced by maintaining the animals on 1% NaCl solution in place of drinking water. Probucol (10 or 30 mg/kg/day), both of which doses are therapeutic in humans was given by gastric gavage once daily to salt-loaded SHRSP. Animals receiving vehicle were used as controls. Probucol did not influence the elevation of blood pressure. Although probucol did not improve the survival rate of salt-loaded SHRSP, 30 mg/ kg/day of probucol slightly but significantly delayed the development of neurological events (p=0.0235 by generalized Wilcoxon test). However, a high dose of probucol (100 mg/kg/day) did not change the survival or neurological events of salt-loaded SHRSP. These results suggest that probucol may be protective against the development of neurological events, but is not preventive for the progression of stroke in SHRSP. PMID- 11016806 TI - Effects of monosodium glutamate-induced obesity in spontaneously hypertensive rats vs. Wistar Kyoto rats: serum leptin and blood flow to brown adipose tissue. AB - We compared the effects of hypothalamic obesity induced by neonatal monosodium glutamate (MSG) treatment between spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Newborn WKY and SHR were injected intraperitoneally with 4 mg/kg body weight of MSG daily for 5 days. At 6 months of age, the obesity of SHR was more advanced than that of WKY, but at 14 months of age the severity of obesity was similar between the two strains. Hypertriglyceridemia was enhanced in MSG-treated SHR as compared with MSG-treated WKY. Systolic blood pressure measured by the tail-cuff method was consistently lower in MSG-treated SHR than in control SHR, whereas blood pressure was not affected by neonatal MSG treatment in WKY. Food restriction reduced body weight more in control SHR than in control WKY, with the former also showing enhanced ketogenesis. Neonatal MSG treatment abolished the accelerated reduction of body weight in SHR. Serum leptin concentration was markedly increased in MSG-treated obese rats, though no differences were seen between WKY and SHR in the control or MSG-treated groups. Serum leptin was closely correlated with both Lee obese index and mesenteric fat weight over the strain. Blood flow in interscapular brown adipose tissue (BAT) measured by Laser Doppler flowmetry was significantly increased in response to beta3-adrenoceptor agonist BRL26830A in both the control and MSG-treated rats. However, the response of blood flow was not affected by MSG treatment or strain difference. The present study demonstrated some strain differences in response to neonatal MSG treatment between WKY and SHR. These differences could not be explained by the difference in serum leptin level or beta3-adrenergic reactivity in BAT. PMID- 11016807 TI - Roles of renal dopamine and kallikrein-kinin systems in antihypertensive mechanisms of exercise in rats. AB - We have previously shown that both renal dopamine (DA) and kallikrein-kinin systems are activated by exercise in mild hypertensives. We aimed to confirm the effects of exercise on the renal DA system and the stimulatory effects of DA on the renal kallikrein-kinin system in rats. In experiment 1, 12 male Dahl salt sensitive (DS) rats given a 4% salt diet were divided into two groups. Rats in the exercise group were forced to run at 8 m/min, 60 min/day, 5 days/week for 4 weeks. Daily urinary volume, urinary excretion of sodium, free DA, and kallikrein activity were measured weekly. Renal aromatic-L-amino-acid decarboxylase (AADC) activities were assayed at the end of the experiment. In experiment 2, 15 male Sprague-Dawley (SD) rats were randomly divided into 3 groups, a DA-5 (5 microg of DA/kg/min), a DA-10 (10 microg of DA/kg/min), and a control group. DA or vehicle was administered subcutaneously with an osmotic pump for 2 weeks. Daily urinary volume, urinary excretion of sodium, aldosterone, DA, and kallikrein activity were measured weekly. Plasma renin activity, aldosterone concentration, and renal kallikrein mRNA levels were determined at the end of the experiment. In experiment 1, urinary excretion of free DA and renal AADC activities in the exercise group were significantly higher than those in the non-exercise group at week 4. In experiment 2, renal kallikrein mRNA levels and urinary volume were significantly increased in the DA-10 group compared to the control group, although there were no differences in urinary kallikrein activities. Plasma aldosterone concentration was significantly decreased in the DA-10 group compared to that in the control group despite a lack of differences in plasma renin activities. In conclusion, exercise increased the urinary excretion of free DA, probably through increased renal AADC activity in DS rats. DA amplified renal kallikrein mRNA levels and decreased plasma aldosterone levels, probably through its suppression of aldosterone in the adrenal glands. Activation of the kallikrein-kinin system might be counteracted by post-transcriptional modification of aldosterone. These results suggest that exercise enhances renal dopamine production by activating renal AADC activity, which in turn stimulates the renal kallikrein-kinin system. PMID- 11016808 TI - The effects of calcium channel blockers on nuclear factor kappa B activation in the mesangium cells. AB - It has been reported that calcium channel blockers (CCBs) have an inhibitory action on cell growth and transcriptional changes induced by cytokines and hormones. In this study, we examined the effects of CCBs on nuclear factor kappa B (NFkappaB), which plays a key role in the intracellular signaling of various growth factors and cytokines. The activity of NFkappaB was determined by luciferase assay with the transfection of the reporter gene, which has six NFkappaB-recognizing sequences in the upstream of herpes simplex virus thymidine kinase promoter. In cultured human mesangial cells, increased intracellular calcium concentration by calcium ionophore, A23187, showed a stimulatory effect on the phorbor 12-myristate 13-acetate (PMA)-induced activation of NFkappaB, while L-type calcium channel agonist, Bay K 8644, did not have any significant effects on either basal or PMA-stimulated activity of NFkappaB. At a higher concentration (10 microM), nifedipine, verapamil, or efonidipine showed an inhibitory effect on the activation of NFkappaB by PMA and A23187, while at a lower concentration (1 microM), only efonidipine showed a significant inhibitory effect. From these results, we conclude that CCBs have an inhibitory effect on NFkappaB via the independent pathway of an L-type calcium channel and that the potency of this effect is variable among L-type calcium channel blockers. PMID- 11016810 TI - The effects of verapamil SR and bisoprolol on reducing the sympathetic nervous system's activity. AB - To assess the response of the sympathetic nervous system (SNS) to the handgrip test in essential hypertensive patients and to evaluate the effects of verapamil SR and bisoprolol on the reduction of the SNS's activity. Seventy eight essential hypertensive patients (50 receiving verapamil SR treatment and 28 receiving bisoprolol treatment) took the handgrip test while the SBP, DBP, and HR were measured on three occasions during the test (before test, 3 min after the patients squeezed the handgrip, and 2 min after the handgrip was released). Before and after the patients received Verapamil SR or Bisoprolol treatment, the plasma concentrations of epinephrine(E), norepinephrine (NE), angiotensin-II (AII), aldosterone (ALD), endothelin-1 (ET-1) and renin activity (RA) were measured post-test. 1) In about 70% of the essential hypertensive patients, SNS activity was above normal. Their HR and BP exceeded 20% when responding to stress. 2) In these patients, the baseline plasma concentrations of E, NE, AII, ET-1, ALD, and RA were higher than those whose SNS's activity was normal. 3) After 6 weeks of treatment, all the patients' BPs decreased remarkably. Verapamil SR could reduce the plasma concentrations of NE, AII, and ET-1 and increase RA. Bisoprolol could reduce E and RA. These two antihypertension drugs can both decrease BP and reduce the activity of SNS through different mechanisms. PMID- 11016809 TI - Effects of vasodilatory antihypertensive agents on endothelial dysfunction in rats with ischemic acute renal failure. AB - Ischemic acute renal failure is associated with vascular endothelial dysfunction. We examined whether vasodilatory antihypertensive agents would improve endothelial function in rats with ischemia/reperfusion renal injury. Rat kidneys were isolated and perfused after clipping of the bilateral renal arteries for 45 min and reperfusion for 24 h, and renal perfusion pressure and nitric oxide concentration in the venous effluent (chemiluminescence assay) were monitored. Preischemic administration of celiprolol (a beta-blocker; 100 mg/kg p.o.), benidipine (a calcium channel blocker; 1 mg/kg p.o.), or imidapril (an angiotensin converting-enzyme inhibitor; 3 mg/kg p.o.) restored endothelial function in rats subjected to acute renal ischemia (deltarenal perfusion pressure [10(-8) M acetylcholine]: sham -42+/-3%, ischemia -31+/-1%, ischemia +celiprolol 39+/-1%*, ischemia+benidipine -38+/-2%*, ischemia+imidapril -42+/-2%*; *p<0.05 vs. ischemia). Serum urea nitrogen and creatinine levels were also lower in the treated groups. Furthermore, ischemia-induced decreases in the response to acetylcholine and renal excretory function were smaller in SHR than in deoxycorticosterone-salt hypertensive rats, in which endothelial damage was marked. These results suggest that preischemic endothelial function may influence the degree of ischemic renal injury. Calcium channel blockers, converting-enzyme inhibitors, and endothelial NO synthase-activating beta-blockers had beneficial effects on renovascular endothelial dysfunction due to ischemia. PMID- 11016811 TI - Plasma endothelin-1 levels and circulating endothelial cells in patients with aortoarteritis. AB - To investigate the correlation between plasma endothelin-1 (ET-1), circulating endothelial cells (CECs), and the disease activity in patients with aortoarteritis. In this study, radioimmunoassay was used to measure plasma levels of ET-1 in 56 patients with aortoarteritis. Circulating endothelial cell counts were also carried out as an indicator of vessel wall lesions. The plasma levels of ET-1 and CECs in the active disease patient group were significantly higher than those in inactive patient group (p<0.001). A significant positive correlation was found between plasma ET-1 levels and erythrocyte sedimentation rates (ESRs) in patients with aortoarteritis (r=0.645, p<0.001), as well as CECs (r=0.876, p<0.001). These results suggested that the ET-1 secreted during the active stages of aortoarteritis may cause constriction and proliferation of vascular smooth muscle cells, thus contributing to the pathogenesis of luminal narrowing. The increased CECs might serve as a marker of disease activity. PMID- 11016812 TI - Endogenous digitalislike factor: an update. AB - This issue of Hypertension Research contains the review and original articles presented at the International Symposium on Natriuretic and Digitalis-Like Factors held in Chitose, Hokkaido, Japan, on August 24, 1999. The symposium was the satellite meeting of the 9th International Conference on the Na/K-ATPase and Related ATPases, which was held in Sapporo, Hokkaido. At the symposium, it became clear that ouabain is the most promising candidate for a circulating hormone to regulate a number of physiological functions, including hypertension, and that other minor substances may also exist as endogenous digitalislike factors. Most of the symposium contributors submitted papers to this journal. I am going to summarize briefly the research history and current research results on endogenous digitalislike factors (EDLF). PMID- 11016813 TI - Putative roles of ouabainlike compound in hypertension: revisited. AB - It is clear that defective renal sodium handling plays an important role in the development of hypertension and that this abnormality could be caused by heterogeneous hereditary factors in the kidney. It is likely that sodium pump inhibitors with or without whole-body autoregulation gradually produce a rise in blood pressure in response to retained body sodium. Accumulated evidence has suggested that several sodium pump inhibitors similar to cardiotonic steroids are present in the human body. Ouabainlike compound (OLC) has been found to be increased with high sodium intake and hypervolemia, and in essential hypertension, mineralocorticoid hypertension, and pregnancy-induced hypertension. Further, blocking the action of OLC with digibind or a novel anti-ouabain agent has been observed to lower blood pressure in several models of experimental and clinical hypertension. The blockade of OLC action may become the basis of novel rational antihypertensive agents and may help to solve the problems still present in the management of hypertensive patients. PMID- 11016814 TI - PST 2238: a new antihypertensive compound that modulates the Na-K pump 'in vivo' and 'in vitro'. AB - A primary renal alteration due to a genetic polymorphism of the cytoskeletal protein adducin associated with an up-regulation of the renal Na-K pump and increased levels of ouabainlike factor (OLF) has been identified as a possible causes of hypertension in Milan rats (MHS). This adducin polymorphism has also been found to be associated with hypertension and the blood pressure changes related to renal Na handling in humans and increased OLF levels have been found in a relevant portion of hypertensive patients. Increased activity and expression of the Na-K pump has also been observed under the following 'in vitro' and 'in vivo' conditions: rat renal cells transfected with the 'hypertensive' variant of adducin, as compared with normal cells; normal rat renal cells incubated for 5 days with 10(-9) M ouabain and normal rats made hypertensive by a chronic infusion of low doses of ouabain (OS rats). An up-regulation of the Na-K pump seems therefore to be a common biochemical alteration induced both by an adducin polymorphism and/or chronic exposure to low concentrations of ouabain (or OLF). A new antihypertensive compound, PST 2238, that selectively antagonizes the pressor effect and the alteration of the renal Na-K pump induced both by an adducin polymorphism and OLF, is described. The ability of PST 2238 to lower blood pressure and normalize the Na-K pump both in MHS and OS rats suggests that this compound could be useful in the treatment of those forms of essential hypertension in which renal Na-handling alterations are associated with either adducin polymorphisms and/or increased OLF levels. PMID- 11016815 TI - Liquid chromatography mass spectrometric analysis of ouabainlike factor in biological fluid. AB - Ouabainlike factor (OLF), assayed as ouabainlike immunoreactivity (OLI), is a probable endogenous digitalislike factor (EDLF). Liquid chromatography/mass spectrometry (LC/MS) is one of the most highly sensitive tools for obtaining structural information regarding low-molecular weight materials in a target compound, and to measure the concentrations of these materials. We have previously reported that OLI can be isolated from the culture supernatant of the rat pheochromocytoma cell line, PC12, by several reverse-phase chromatography and LC/MS techniques. The present study was performed to characterize OLF from biological fluids such as plasma and culture supernatant of PC12 cells by LC/MS. The previous applications of LC/MS to OLI in plasma have been limited to structural identification at the final stages of isolation, in which the starting volume of plasma has been over 10 I. In the present study, we tried to minimize the volume of plasma, and to develop a new preclearing step to gain adequate LC/MS characterization using MS/MS analysis. The plasma was acidified, and OLI was purified by ODS column chromatography. OLI in chromatographic fractions from plasma was assayed by a sensitive enzyme-linked immunosorbent assay for ouabain. After Sep-Pak treatment and two rounds of ODS column chromatography, OLI was identified from 80 ml of plasma. The structure of the purified OLI was identical to authentic ouabain and digoxin, as assessed by LC/MS. In conclusion, we identified the chemically or structurally clarified ouabain and digoxin as the circulating form in plasma by LC/MS. PMID- 11016816 TI - Analysis of human ouabainlike compound by micellar electrokinetic chromatography. AB - In this preliminary study we have optimised micellar electrokinetic chromatography (MEKC, a form of capillary electrophoresis) to enable the chromatographic and spectral characteristics of human ouabainlike compound (OLC) to be investigated. Sera from fifty patients were combined to form a pool (100 ml) whilst urine (92.5 ml) was obtained from a normal healthy volunteer. Both samples were initially concentrated and partially purified by solid phase extraction before further purification by sequential HPLC separations. Final volumes for both extracts were 100 microl. MEKC was performed on a HP(3D) CE instrument with voltage set at 20 KV, capillary temperature at 20 degrees C, injection time 4 s, sample volume 10 nl, with detection by photodiode array. A compound was found in both serum and urine that had similar elution and spectral characteristics to authentic ouabain. We conclude that MEKC is potentially a useful tool for the analysis of human OLC. PMID- 11016817 TI - The structure of the digitalislike and natriuretic factors identified as macrocyclic derivatives of the inorganic carbon suboxide. AB - The Natriuretic and Endogenous DigitalisLike Factors (EDLFs) are disclosed to be cyclomeric and macroring closed derivatives of the inorganic carbon suboxide. The macrocyclic cyclohexamer with six carbon suboxide units has a molar mass of 408.2 Da, as previously been found for the EDLF of animal origin. The anhydrous cyclohexameric factor is lipophilic but is transformed into more hydrophilic derivatives by the stepwise addition of water. Based on the present findings, it appears that EDLFs exist in solution as an equilibrium mixture of lipophilic and hydrophilic forms and not as a single chemical substance. This structural assumption better accounts for the earlier observed highly anomalous properties of EDLFs. The simultaneously found higher molar mass (4,100 and 4,900 Da) macrocyclic carbon suboxide derivatives are tentatively identified as the Natriuretic factors. PMID- 11016818 TI - Receptor occupancy with digoxin vs receptor occupancy with a putative endogenous digitalislike factor. AB - The possibility that an endogenous ligand for the digitalis receptor might exist has been a source of speculation resulting in efforts over the past decades to identify such a hormone. In the current context it is of interest that prolonged wash of myocardial and skeletal muscular samples from subjects who were not in digoxin treatment generally resulted in small tendencies to increase 3H-ouabain binding ranging from 2 to 9% and from -2 to 7%, respectively. It may be appreciated that neither wet weight nor water content of left ventricular or skeletal muscular samples have been found to change as a result of the prolonged wash. Although these tendencies most likely are a mere play of chance, it may be argued that the studies did not entirely rule out the possibility of the existence of a quantitatively small amount of endogenous digitalislike factor. However, based on the evaluations of receptor occupancy with digoxin during treatment of 24-34% in the left ventricle and 9-13% in skeletal muscle, it would seem reasonable to expect that a comparatively larger fraction of digitalis receptors should be occupied by a putative endogenous digitalislike factor, if such a factor were to be of any physiological significance. PMID- 11016819 TI - Binding of ouabain and human ouabainlike substance to different Na+, K+-ATPase isoforms. AB - There is very little on the affinity of the human immunoreactive ouabainlike substance (OLS) to individual alpha-isoforms of Na+,K+-ATPase. The present study addresses this issue by comparing ouabain and OLS binding to dog kidney alpha1, rabbit kidney alpha1 and porcine cerebral cortex alpha3 Na+,K+-ATPase. OLS was initially isolated by solid phase extraction from human serum using C18 columns. The extract was further purified by reverse phase HPLC in an acetonitrile/water (containing 0.1% TFA) step-up gradient (16-80%). In this system, two distinct ouabain immunoreactive peaks were resolved. Peak I demonstrated a polarity identical with that of authentic ouabain. In contrast, peak II was relatively non polar and eluted later in the run. The final step in the purification of OLS involved immuno-affinity chromatography of peak I using a specific sepharose immobilized mouse monoclonal anti-ouabain antiserum. Dose response curves (range 0-100 nmol/l) for ouabain with canine alpha1 and porcine alpha3 Na+,K+-ATPase showed similar inhibitory profiles (IC50=15 nmol/l), whilst rabbit alpha1 Na+,K+ ATPase was relatively insensitive to ouabain and purified peak I OLS. Two fold serial dilution of Peak I OLS, with subsequent analysis by canine and porcine Na+,K+-ATPase inhibition assays and RIA, demonstrated strong positive correlations between OLS determined by RIA and both canine (y=0.945x-2.532, r2=0.977) and porcine (y=0.428x-1.685; r2=0.993) Na+,K+-ATPase assays. The difference in the respective slopes suggests, however, that peak I OLS has a greater affinity for the canine derived enzyme compared to the porcine. In conclusion, these data suggest that like authentic ouabain, peak I OLS is a isoform and species selective. PMID- 11016820 TI - Digitalis and digitalislike compounds down-regulate gene expression of the intracellular signaling protein 14-3-3 in rat lens. AB - Na+,K+-ATPase activity in the epithelial layer is fundamental to the maintenance of ionic concentration gradients and transparency of the lens. Recently we have identified endogenous digitalislike compounds (DLC), 19-norbufalin and its peptide derivatives, in human cataractous lenses (Lichtstein et al. Eur J Biochem 216: 261-268, 1993). Lenses were treated with 10 nM ouabain, bufalin or 19 norbufalin derivative for 24 h and were compared to control lenses. Differential display analysis revealed that one of the down-regulated genes was 14-3-3 theta. Down-regulation was confirmed by Northern blot and by RT-PCR analysis. RT-PCR of additional 14-3-3 isoforms revealed that the eta and gamma isoforms of 14-3-3 are also down-regulated by ouabain, bufalin and 19-norbufalin derivative, whereas the zeta isoform is down-regulated only by bufalin. These results demonstrate that one of the consequences of Na+,K+-ATPase inhibition by exogenous or endogenous inhibitors is the down-regulation of mRNA transcripts encoding several isoforms of 14-3-3. Since the 14-3-3 proteins are multifunctional regulatory proteins, the reduction in the abundance of various isoforms will have profound effects on cell function. Furthermore, These results, together with the demonstration of digitalislike compounds in the normal lens, and their increased level in human cataractous lenses, strongly suggests their involvement in the molecular mechanisms responsible for cataract formation. PMID- 11016821 TI - Differential regulation of the sodium pump alpha-subunit isoform gene by ouabain and digoxin in tissues of rats. AB - The effects of ouabain and digoxin on both the systolic blood pressure (SBP) and sodium pump alpha-subunit expression in some tissues of rats were compared. Normal rats were injected with ouabain, digoxin, and normal saline (NS), respectively, everyday, and indirect SBP was recorded once a week. Six weeks later, all the rats were killed, and sodium pump alpha1-, alpha2-, and alpha3 subunit mRNA levels were detected in the myocardium, kidney, adrenal gland, aortic smooth muscle, and hypothalamus by the RT-PCR method. The results showed that the SBP of rats infused with ouabain increased significantly at the end of week 6, while no difference in SBP was found between the digoxin and NS groups. The effects of ouabain and digoxin on sodium pump alpha-subunit isoform expression were also different. Myocardium: both ouabain and digoxin stimulated expression of the alpha3-isoform whereas alpha2 was unchanged. Levels of the alpha1 isoform decreased significantly in the ouabain group and decreased slightly in the digoxin group, respectively. Kidney: digoxin had the same effects as ouabain. alpha1 levels increased, but those of alpha2 and alpha3 remained unchanged. Adrenal gland: alpha2 and alpha3 levels increased, but those of alpha1 decreased in the ouabain group. alpha1 and alpha3 levels increased and those of alpha2 remained unchanged in the digoxin group. Aortic smooth muscle: both ouabain and digoxin increased alpha1 and alpha3 expression. alpha2 levels decreased in the digoxin group but remained unchanged in the ouabain group. Hypothalamus: both ouabain and digoxin stimulated alpha1 expression, while alpha2 and alpha3 levels remained unchanged. The results of this study have shown that ouabain and digoxin have the different effects on both the systolic blood pressure and expression of sodium pump alpha-subunit isoforms in some tissues in rats. Further studies on the expression of sodium pump alpha-subunit isoforms might be helpful for the understanding of the physiological role of endogenous ouabain and the molecular mechanisms involved in the pathogenesis of hypertension. PMID- 11016822 TI - Effects of endogenous ouabain on the development of hypertension in 1k1c hypertensive rats. AB - This study was designed to evaluate the role of endogenous ouabain (EO) in the development of hypertension in 1ktc (one kidney, one clip) hypertensive rats. First, the EO content of the serum of 1k1c hypertensive rats and normal Sprague Dawley (SD) rats was detected by the enzyme linked immunosorbent assay method (ELISA). Second, blood pressure changes in the 1k1c rats were recorded directly after the 1k1c rats were injected randomly with anti-ouabain antibody, normal rabbit IgG, and normal saline, respectively. The results showed that EO levels in the serum of 1k1c hypertensive rats were significantly higher than those of normal SD rats (2.25 +/-0.92 microg/l vs. 1.12 +/- 0.17 microg/l, p< 0.01), and correlated significantly with systolic blood pressure (r= 0.59, p< 0.05). Anti ouabain antibody was able to significantly decrease the blood pressure of 1k1c hypertensive rats in a dose-dependent manner, while normal rabbit IgG or normal saline was not. These results indicate that endogenous ouabain might play an important role in the development of hypertension in 1k1c hypertensive rats. PMID- 11016824 TI - Chronic hypertension induced by ouabain but not digoxin in the rat: antihypertensive effect of digoxin and digitoxin. AB - Elevated circulating levels of an endogenous ouabain (EO) have been associated with essential hypertension. To investigate structure-activity relationships relevant to blood pressure, we infused either ouabain, ouabagenin, digoxin or digitoxin at 30 microg/kg/day in normal Sprague Dawley rats. After five weeks, the ouabain and ouabagenin infused rats were hypertensive, whereas blood pressures declined below their vehicle controls in rats infused with digoxin or digitoxin. In a second study, mean blood pressures were 118.5+/-1.7 mmHg in rats infused with ouabain (15 microg/kg/day) on day 35 vs. 98.3+/-1.8 and 100.3+/-1.1 mmHg in the digoxin (30 microg/kg/day) and vehicle infused groups (both p<0.005 vs. ouabain), respectively. Plasma and kidney levels of ouabain immunoreactivity were increased 4-8 fold in ouabain infused rats while blood pressure and plasma levels of ouabain returned to normal one week following discontinuation of the steroid infusion. In rats with ouabain-dependent hypertension, secondary infusions of digoxin or digitoxin (30 microg/kg/day) normalized blood pressure even though circulating ouabain remained elevated. In digoxin infused rats, neither blood pressure nor kidney digoxin immunoreactivity was raised whereas plasma digoxin was increased. Collectively, the results show that the hemodynamic effects of these sodium pump inhibitors differ dramatically during prolonged administration and that tissue rather than circulating levels of these agents appear to better explain their effects on blood pressure. These studies suggest that sodium pump inhibition is not the exclusive mediator of the hemodynamic effects of these cardiac glycosides and demonstrate the presence of structure specific mechanisms that regulate their tissue levels and effects on long-term blood pressure. PMID- 11016825 TI - Endogenous digitalislike factors in obstructive sleep apnea. AB - Recent studies have provided evidence that hypoxia may stimulate the release of endogenous digitalislike factors (EDLF). Obstructive sleep apnea (OSA) is characterized by intermittent hypoxia during sleep and may be associated with sympathetic activation and a high risk of developing hypertension. This study was designed to measure EDLF in the plasma of patients with OSA diagnosed by polysomnography, with patients being classified by the number of apneic-hypopneic episodes/h sleep (apnea-hypopnea index, AHI). Plasma was obtained in the morning from 8 male normotensive OSA patients (OSA-N) (AHI 70+/-6), 2 untreated hypertensive OSA patients (OSA-HT), and 11 age-matched healthy male controls (C). EDLFs of different hydrophobicities were separated from the same plasma sample by solid-state C18-cartridges with 25% acetonitrile (ACN) (EDLF-1) followed by 40% ACN (EDLF-2). This procedure recovered ouabain in the first fraction and digoxin and digoxigenin in the second. EDLF was quantified in pM ouabain-equivalents by a human placenta radioreceptor assay. EDLF-1 levels were similar for OSA-N and C (231+/-55 vs. 258+/-58), whereas EDLF-2 levels were increased in OSA-N (244+/-51 vs. 110+/-25 in C, p=0.02). Norepinephrine was increased in apneics. The two OSA HT had EDLF and norepinephrine levels similar to OSA-N. These preliminary results suggest that OSA is associated with an increase in the more hydrophobic EDLF levels in both normotensive and hypertensive states. No significant increase was found for the less hydrophobic ouabain-like EDLF. PMID- 11016823 TI - Different effects of in vivo ouabain and digoxin on renal artery function and blood pressure in the rat. AB - To investigate vascular mechanisms in hypertension, we isolated renal arterial rings from rats with ouabain-dependent hypertension and studied their function. In rats infused with ouabain for 5 weeks, systolic and mean blood pressures (BP) were increased relative to controls. Contractions evoked by high KCl solutions were greater in rings from ouabain-infused rats whereas the threshold concentrations and EC50s for KCl and the peak caffeine contractures were not different. KCl contractures were not affected by 5 microM prazosin. Phenylephrine contractures were increased marginally in ouabain-infused rats, while acetylcholine-induced relaxation was normal. In vitro superfusion of rings with 10 nM ouabain or digoxin did not affect the measured parameters. Plasma ouabain, BP, and all evoked responses were normal one week following interruption of the ouabain infusion. In a second study, BP increased in ouabain (15 microg/kg/day, n= 23), but not digoxin (30 microg/kg/day, n=12), or vehicle-infused (n=16) rats. KCl contractures were greater in rings from ouabain-but decreased in rings from digoxin-infused rats, respectively and correlated with systolic and mean BP (r=0.69, n=30, p<0.005). Peak caffeine (25 mM) responses were similar but the area under the contraction was reduced in the vessels from ouabain-infused rats and correlated inversely with MBP (r=-0.47, n=33, p<0.02). We conclude that a voltage-dependent component of tone in the rat renal artery is reversibly and specifically augmented by in vivo administration of ouabain whereas it is diminished by in vivo digoxin. Vascular production of and response to nitric oxide does not appear to be impaired in the ouabain model. Alterations of intracellular Ca2+ storage and Ca2+ influx in response to in vivo ouabain may underlie the increase in renal vascular resistance and hypertension in this model. The opposite effects of ouabain and digoxin on the hemodynamic and vascular parameters in this study indicate that these agents have novel mechanisms of action in vivo that may not be mediated exclusively by sodium potassium pumps. PMID- 11016826 TI - Endogenous ouabain and its binding globulin: effects of physical exercise and study on the globulin's tissue distribution. AB - Ouabain, that has been isolated from bovine adrenals and hypothalamus, is a new cardiotonic steroid hormone, which is either synthesized in the adrenals or stored there after it has absorbed from the diet. Little is known in vivo which events may lead to the release of ouabain into blood. Moreover, a binding protein for cardiotonic steroids exists in blood, which binds cardiac glycosides with high affinity. It may affect the action of endogenous ouabain on heart and circulation, but the physiological function of this protein is unclear. To realize, which physiological stimuli in vivo may affect blood concentrations of endogenous ouabain and which function the cardiotonic binding protein may have in modulating ouabain effects, the effect of physical exercise on endogenous ouabain was studied and the tissue distribution of its binding protein was investigated. We found that endogenous ouabain changes rapidly in blood upon physical exercise and behaves like expected for a hormone of circulation. The cardiotonic steroid binding globulin shows the highest concentration in the kidney, which suggests that sodium pumps of the kidney are protected against its inhibition by ouabain which would lead not only to natriuresis but also to a deleterious loss of glucose, amino acids and phosphate. PMID- 11016827 TI - The rice Rim2 transcript accumulates in response to Magnaporthe grisea and its predicted protein product shares similarity with TNP2-like proteins encoded by CACTA transposons. AB - A rice transcript, Rim2, was identified that accumulated in both incompatible and compatible interactions between rice and Magnaporthe grisea. The Rim2 transcript also accumulated in response to treatment with a cell wall elicitor derived from M. grisea. A 3.3-kb RIM2 cDNA clone was isolated and is predicted to encode a protein of 653 amino acids, which shares 32 55% identity with TNP2-like proteins encoded by CACTA transposons of other plants. A 1.05-kb segment of the Rim2 sequence shows 82% nucleotide sequence identity with sequences flanking the A1 and C members of the rice Xa21 disease resistance gene family. The 5'-upstream region of Rim2 was cloned and the transcriptional start sites were identified. The 5' and 3' noncoding termini of Rim2 are AT-rich. A cis-element showing similarity to a sequence that mediates defense-associated transcriptional activation of the tobacco retrotransposon Tnt1, and four motifs that fit the consensus sequence of the elicitor-responsive elements in the promoters of the parsley PR-1 genes were found in the 5'-upstream region. Four imperfect tandem repeats were identified in the 3' noncoding terminus. Southern analysis with genomic DNA from different rice species indicated that Rim2 is present in 3-4 copies per genome. These results suggest that Rim2 may be one component of a large CACTA-like element, whose transcript accumulates in response to attack by pathogens. PMID- 11016828 TI - Identification and characterisation of a silkworm ABC transporter gene homologous to Drosophila white. AB - In the silkworm, Bombyx mori, many eye- and egg-colour mutations affecting the synthesis and accumulation of ommochrome pigments have been described. In order to understand the pigment precursor transporters involved, ABC transporter genes homologous to the Drosophila white gene were isolated from the silkworm. Reverse transcriptase-polymerase chain reactions (RT-PCR) using embryonic mRNA amplified three cDNA fragments, named Bmwh1, Bmwh2 and Bmwh3 that showed homology to the white gene. Since Bmwh3 shows the highest degree of sequence identity and a similar expression pattern to the Drosophila homologue, we characterised this gene further. A 2667-bp Bmwh3 cDNA isolated from an embryonic library has one ORF encoding a polypeptide of 687 amino acids. The predicted protein has one ATP binding domain, six transmembrane-spanning segments and high similarity to the Drosophila WHITE protein. Southern analysis indicates that Bmwh3 is a single-copy gene. Polyadenylated Bmwh3 transcripts about 2.7 kb long were detected in eggs, Malpighian tubules and pupal heads, but not in testes, posterior silk glands or fat body cells. The level of Bmwh3 mRNA was reduced in w3 and w3ol mutants but normal in other egg- and eye-colour mutants, suggesting that Bmwh3 correspond to the w3 locus. Genetic analysis was used to map the cloned gene to chromosome 10. PMID- 11016829 TI - Transgene repeats in aspen: molecular characterisation suggests simultaneous integration of independent T-DNAs into receptive hotspots in the host genome. AB - Rearrangements of T-DNAs during genetic transformation of plants can result in the insertion of transgenes in the form of repeats into the host genome and frequently lead to loss of transgene expression. To obtain insight into the mechanism of repeat formation we screened 45 transgenic lines of aspen and hybrid aspen transformed with six different gene constructs. The frequency of T-DNA repeat formation among randomly screened transgenic lines was found to be about 21%. In ten transgenic lines direct repeats were detected. An inverted repeat was found in one other transgenic line. Sequencing of the junctions between the T-DNA inserts revealed identical residual right-border repeat sequences at the repeat junctions in all ten transgenic lines that had direct repeats. Formation of "precise" junctions based on short regions of sequence similarity between recombining strands was observed in three transgenic lines transformed with the same plasmid. Additional DNA sequences termed filler DNAs were found to be inserted between the T-DNA repeats at eight junctions where there was no similarity between recombining ends. The length of the filler DNAs varied from 4 to almost 300 bp. Small filler DNAs--a few base pairs long--were in most cases copied from T-DNA near the break points. The large filler sequences of about 300 bp in two transgenic lines were found to be of host plant origin, suggesting that transgene repeat formation occurred as a result of the simultaneous invasion of a receptive site in the host genome by two independent T-DNA strands. On the basis of the results obtained, and in the light of previous reports on T-DNA/plant DNA junctions in aspen and other crop plants, a mechanistic model for transgene rearrangement and filler formation is suggested. PMID- 11016830 TI - Isolation and characterisation of a mutation in the PMR1 gene encoding a Golgi membrane ATPase, which causes hypersensitivity to over-expression of Clb3 in Saccharomyces cerevisiae. AB - We screened for mutant strains of Saccharomyces cerevisiae that are sensitive to overexpression of specific cyclins, and identified mutations in two genes that caused growth inhibition in response to mild overexpression of Clb3. One was the ANP1 gene, which encodes a glycosyltransferase previously identified by a similar strategy using Clb2 instead of Clb3. This paper describes the second strain of S. cerevisiae that is hypersensitive to Clb3 expression. The gene mutated in this strain was identified as PMR1, which encodes a Ca2+-ATPase located in the Golgi membrane. The protein product of pmr1-1 was truncated at residue 409 and thus lacked the C-terminal ATPase domain. The pmr1-1 strain was hypersensitive to over expression of Clb3, but not Cln2, Clb5 or Clb2. The lethality due to Clb3 expression in pmr1-1 could be suppressed by adding Ca2+ ions to the medium. The pmr1-1 strain proved to be defective in glycosylation, and the defects in glycosylation were exacerbated by high levels of Clb3. On induction of Clb3 expression in the pmr1-1 strain, the cells arrested at anaphase with an elongated daughter bud. We discuss possible interpretations of this synthetic lethal phenotype. PMID- 11016831 TI - Overexpression of the catalytic subunit of DNA polymerase gamma results in depletion of mitochondrial DNA in Drosophila melanogaster. AB - The mechanisms involved in the regulation of mitochondrial DNA (mtDNA) replication, a process that is crucial for mitochondrial biogenesis, are not well understood. In this study, we evaluate the role of DNA polymerase gamma (pol gamma), the key enzyme in mtDNA replication, in both Drosophila cell culture and in developing flies. We report that overexpression of the pol gamma catalytic subunit (pol gamma-alpha) in cultured Schneider cells does not alter either the amount of mtDNA or the growth rate of the culture. The polypeptide is properly targeted to mitochondria, yet the large excess of pol gamma-alpha does not interfere with mtDNA replication under these conditions where the endogenous polypeptide is apparently present in amounts that exceed of the demand for its function in the cell. In striking contrast, overexpression of pol gamma-alpha at the same level in transgenic flies interferes with the mtDNA replication process, presumably by altering the mechanism of DNA synthesis, suggesting differential requirements for, and/or regulation of, mtDNA replication in Drosophila cell culture versus the developing organism. Overexpression of pol gamma-alpha in transgenic flies produces a significant depletion of mtDNA that causes a broad variety of phenotypic effects. These alterations range from pupal lethality to moderate morphological abnormalities in adults. depending on the level and temporal pattern of overexpression. Our results demonstrate that although cells may tolerate a variable amount of the pol gamma catalytic subunit under some conditions, its level may be critical in the context of the whole organism. PMID- 11016832 TI - Retroelements contribute to the excess low-copy-number DNA in pine. AB - Excess DNA in the single-copy component is rarely recognized as a contributor to the C-value paradox yet the single-copy component of the pine genome is reported to comprise over 3000 Mb of DNA, in large excess over the estimated 100 Mb required for gene expression. Two hypotheses regarding the factors that might contribute to the excess low-copy-number DNA were tested. The first hypothesis proposes that the excess low-copy kinetic component is actually overestimated by reassociation data analysis. To test this, a previously published C0t curve for Pinus strobus was reanalyzed using a new estimate of genome size based on laser flow cytometry. Part of the excess low-copy-number DNA in the pine genome could be attributed to the choice of parameters used in the analysis of the reassociation data. The second hypothesis holds that diverged retrotransposons contribute to the excess low-copy DNA. Sequences randomly sampled from single copy and low-repetitive kinetic components of the P. taeda genome were characterized. Twelve of 46 fragments cloned from these fractions were found to show sequence similarity to retroelements: hence diverged retroelements contribute to the excess low-repetitive kinetic component in the pine genome. Similarity search was shown to be a conservative method for identifying retroelements, and thus the number of retroelements in the low-copy component was actually underestimated. Most of the retroelements in this fraction were nonfunctional. divergent from known retroelement families and previously reported only for flowering plants. Divergent retrotransposons are thus a major factor contributing to the expansion of the low-repetitive DNA component in higher plants. PMID- 11016833 TI - Effects of HDF1 (Ku70) and HDF2 (Ku80) on spontaneous and DNA damage-induced intrachromosomal recombination in Saccharomyces cerevisiae. AB - The Ku heterodimer binds to the ends of double-stranded breaks (DSBs) in DNA, and is involved in nonhomologous end joining. HDF1 and HDF2, which have been identified in Saccharomyces cerevisiae as homologues of the Ku70 and Ku80 proteins of mammals, reduce radiosensitivity only when homologous recombination repair is impaired and, therefore, affect DSB repair via nonhomologous recombination. Although it has been reported that homologous recombination is defective in the hdf1 null mutant, the roles of HDF1 and HDF2 in this process are not completely clear. We investigated the effect of HDF1 and HDF2 on intrachromosomal recombination by measuring rates of deletion between direct repeats caused by spontaneous and DNA damage-induced events (DEL recombination). We found a decrease in spontaneous DEL recombination in both TCY5 (hdf1delta) and TCY6 (hdf2delta) strains, suggesting that HDF1 and HDF2 play a role in homologous recombination. As DEL recombination events may occur by sister chromatid conversion and/or single-strand annealing, which is initiated by DSBs, HDF1 and HDF2 may be required to recruit proteins to the damaged ends so as to promote single-strand annealing. The strains TCY5 and TCY6 are also defective in methylmethane sulfonate (MMS)- and X-ray-induced, but not in UV-induced DEL recombination. This confirms that HDF1 and HDF2 are required for the completion of DEL recombination by single strand annealing. PMID- 11016834 TI - Up-regulation of genes encoding glycosylphosphatidylinositol (GPI)-attached proteins in response to cell wall damage caused by disruption of FKS1 in Saccharomyces cerevisiae. AB - FKS1 and FKS2 encode alternative catalytic subunits of the glucan synthases that are responsible for synthesis of beta-1,3-glucan in the Saccharomyces cerevisiae cell wall. Disruption of FKS1 reduces the glucan content of the cell wall, increases chitin content and activates the expression of CWP1, which encodes a glycosylphosphatidylinositol (GPI)-dependent cell wall protein. These cellular responses have been regarded as compensating for cell wall damage in order to maintain cell wall integrity. Here, we report the identification, by genome-wide screening, of 22 genes that are transcriptionally up-regulated in fks1delta cells. Among them, five genes were found to encode GPI-attached proteins, three of which are covalently associated with the cell wall. Deletion and replacement analysis of the promoter regions identified Rlm1-binding sequences as being responsible for the up-regulation following disruption of FKS1. Using the rlm1delta tetOp-FKS1 strain, in which the expression of FKS1 can be repressed by doxycycline, we examined the requirement for Rlm1 for the transcriptional up regulation of these five genes. Three of the five genes were not up-regulated by doxycycline, indicating that Rlm1 mediates their up-regulation when FKS1 is inactivated. The remaining two genes were up-regulated by doxycycline, suggesting that a transcription factor other than Rlm1 is involved in their response to disruption of FKS1. PMID- 11016835 TI - Functional analysis of chimeras derived from the Sinorhizobium meliloti and Mesorhizobium loti nodC genes identifies regions controlling chitin oligosaccharide chain length. AB - The rhizobial nodulation gene nodC encodes an N-acetylglucosaminyltransferase that is responsible for the synthesis of chitin oligosaccharides. These oligosaccharides are precursors for the synthesis of the lipo-chitin oligosaccharides that induce cell division and differentiation during the development of nitrogen-fixing root nodules in leguminous plants. The NodC proteins of Mesorhizobium loti and Sinorizobium meliloti yield chitinpentaose and chitintetraose as their main products, respectively. In order to localize regions in these enzymes that are responsible for this difference in product chain length, a set of six chimeric enzymes, comprising different combinations of regions of the NodC proteins from these two bacteria, was expressed in Escherichia coli. The oligosaccharides produced were analyzed using thin-layer chromatography. The major conclusion from this work is that a central 91-amino acid segment does not play any obvious role in determining the difference in the chain length of the major product. Furthermore, the characteristically predominant synthesis of chitintetraose by S. meliloti NodC is mainly dependent on a C-terminal region of maximally 164 amino acids; exchange of only this C terminal region is sufficient to completely convert the M. loti chitinpentaose synthase into an S. meliloti-like chitintetraose synthase. The N-terminal region of 170 amino acids also plays a role in restricting the length of the major product to a tetramer. However, the role of the C-terminal region is clearly dominant, since exchanging the N-terminal region has no effect on the relative amounts of chitintetraose and -pentaose produced when the C-terminal region of S. meliloti NodC is present. The length of a predicted beta-strand around residue 300 in the C-terminal region of various NodC proteins is the only structural element that seems to be related to the length of the chitin oligosaccharides produced by these enzymes; the higher the amount of chitintetraose relative to chitinpentaose, the shorter the predicted beta-strand. This element may therefore be important for the effect of the C-terminal 164 amino acids on chitin oligosaccharide chain length. PMID- 11016836 TI - Premature polyadenylation contributes to the poor expression of the Bacillus thuringiensis cry3Ca1 gene in transgenic potato plants. AB - The cry genes that code for the insecticidal crystal proteins of Bacillus thuringiensis (B.t.) have been widely used to develop insect-resistant transgenic plants. The cry3Ca1 gene has been reported to code for a crystal protein which is particularly potent against the Colorado potato beetle (CPB). To explore the biotechnological potential of cry3Ca1, we introduced this gene into transgenic potato plants under the control of the CaMV 35S promoter. In the resulting transformants, the cry3-Ca1 gene was very poorly expressed. In fact, no full length transcript (2300 nt) could be detected. Instead, only short transcripts of approximately 1100 nt were observed. Analysis of these short transcripts by Northern hybridization, RT-PCR as well as by cloning and sequencing showed that they resulted from premature polyadenylation. These processing events occurred at four sites within the cry3Ca1 coding region (at positions 652, 669, 914 and 981 relative to the translation start site). The sites at which premature polyadenylation took place were not those that showed the highest degree of identity to the canonical AAUAAA motif. Together with other recent data, our findings suggest that premature polyadenylation is an important mechanism which can contribute to the poor expression of transgenes in a foreign host. PMID- 11016837 TI - Genetic analysis of the Saccharomyces cerevisiae Sgs1 helicase defines an essential function for the Sgs1-Top3 complex in the absence of SRS2 or TOP1. AB - The Saccharomyces cerevisiae gene SGS1 encodes a DNA helicase that shows homology to the Escherichia coli protein RecQ and the products of the BLM and WRN genes in humans, which are defective in Bloom's and Werner's syndrome, respectively. Recently, it has been proposed that this helicase is involved in maintaining the integrity of the rDNA and that loss of Sgs1 function leads to accelerated aging. Sgs1 has been isolated on the basis of its genetic interaction with both topoisomerase I and topoisomerase III, as well as in a two-hybrid screen for proteins that interact with the C-terminal portion of topoisomerase II. We have defined the minimal structural elements of Sgs1 required for its interactions with the three topoisomerases, and demonstrate that the complex phenotypes associated with sgs1 mutants are a consequence of a dysfunctional Sgs1-Top3 complex. We also report that the synthetic relationship between mutations in SGS1 and SRS2, which encodes another helicase implicated in recombinational repair, likewise result from a dysfunctional Sgs1-Top3 interaction. Our findings indicate that Sgs1 may act on different DNA structures depending on the activity of topoisomerase I, Srs2 and topoisomerase III. PMID- 11016838 TI - Multiple roles of the Dcdc42 GTPase during wing development in Drosophila melanogaster. AB - The Rho sub-family of GTPases, comprising Rho, Rac and Cdc42. regulates many biological processes, including morphogenesis, cell polarity, migration, the cell cycle and gene expression. It is important to develop genetic approaches to allow the dissection, in vivo, of the mechanisms of GTPase regulation and signal transmission, and their biological consequences. In this regard, wing development in Drosophila melanogaster is an excellent model system. To investigate the functions of the Drosophila Cdc42 GTPase (Dcdc42), we generated phenotypes during wing development, by expression of the dominant-negative N17 and L89 mutants of Dcdc42. We have identified roles for Dcdc42 in wing growth, and in cell fate choice during the development of the wing veins and the peripheral nervous system. Reduction of Dcdc42 signalling following over-expression of Dcdc42N17 resulted in a broader but more diffuse domain characterised by wing-margin sensory bristles. This was correlated with a broadened stripe of wingless expression along the dorsal-ventral boundary of third-instar wing imaginal discs. Together with genetic interactions with loss- and gain-of-function Notch alleles, these data support a role for wild-type Dcdc42 as a negative regulator of Notch signalling. PMID- 11016839 TI - The transposon A(R)4-24P[white, rosy] in Drosophila melanogaster is subject to position-effect variegation at a non-centromeric insertion site. AB - The white gene within the transposon A(R)4-24P[white,rosy] inserted at cytological location 24D1-2 in the euchromatic portion of the Drosophila melanogaster genome exhibits a mosaic pattern of expression which is modified by temperature and Y-chromosome number, as in cases of classical position-effect variegation (PEV). The eye colour of the flies in this variegated stock remains mosaic in the presence of the PEV modifier Su(var)3-6, slightly less so with Su(var)3-9 and Su(var)2-5, and full suppression of variegation occurs in the presence of Su(var)3-7. We have induced further transposition of A(R)4-24 and isolated two mosaic stocks with this transgene at new cytological locations. In these stocks, the A(R)4-24 transposon was flanked by the same genomic DNA fragments as in the original location. Spontaneous loss of these fragments leads to reversion of the variegated eye colour to wild-type. We suggest that the flanking DNA fragments from 24D1-2 are capable of inducing position-effect variegation without any association with centromeric heterochromatin. In situ hybridisation and Southern analysis demonstrate that the 5' flanking genomic fragment contains repeated sequences which are abundantly present in heterochromatin. PMID- 11016840 TI - Isolation and characterization of cDNAs encoding mitochondrial uncoupling proteins in wheat: wheat UCP genes are not regulated by low temperature. AB - Uncoupling proteins (UCP) found in the inner mitochondrial membrane of mammals dissipate the proton electrochemical gradient across the inner membrane to produce heat rather than synthesize ATP. Using PCR-based methods, we isolated two novel cDNA clones, WhUCP1a and WhUCP1b, that encode the mitochondrial uncoupling protein of wheat (Triticum aestivum L.). The cDNA clones each contain one ORF which can code for a protein of 286 amino acids with a predicted molecular mass of about 30.5 kDa, although three amino acid substitutions are found between them. The deduced amino acid sequences each possess three typical mitochondrial carrier signature domains and six membrane-spanning domains which are highly conserved in the mitochondrial transporter family. Southern analysis suggested that the WhUCP1 gene may be present in as many as three copies in the wheat genome, and also that WhUCP proteins may be encoded by a small multigene family. Northern analysis revealed that the steady-state level of the WhUCP1 mRNA is quite low. Quantitative RT-PCR clearly showed that expression of the WhUCP1 gene in wheat seedlings is insensitive to low temperature. Our data suggest that WhUCP1 might have functions other than low temperature-induced thermogenesis, although WhUCP1 possesses all the typical features reported for known UCPs. PMID- 11016841 TI - Characterization of Ce-atl-1, an ATM-like gene from Caenorhabditis elegans. AB - An ATM-like gene was identified in the genome of Caenorhabditis elegans. The putative product of the gene, termed Ce-atl-1 (C. elegans ATM-like 1) consists of 2514 amino acid residues. The C-terminal sequence, which contains a PI-3 kinase like domain, showed good homology with the products of the gene MEC1/ESR1 from budding yeast, the rad3+ gene of fission yeast and mammalian ATM (ataxia telangiectasia and rad3+ related) genes. The results of RNA-mediated interference indicated that the major phenotype associated with repression of Ce-atl-1 was lethality (approximately 50-80%) during early embryogenesis. Among the surviving progeny, males (XO animals) arose at a high frequency (2-30%). In addition, 5% of oocyte chromosomes demonstrated aneuploidy due to a defect in pre-meiotic chromosomal segregation. Gene expression analyses indicated that Ce-atl-1 mRNA was expressed in all larval stages and that its level increased about fivefold in the adult stage. The adult expression level was decreased in the glp-4 mutant, which is defective in germ line proliferation. Ce-atl-1 was strongly expressed in both the mitotic and meiotic cells of adult gonads. In summary, Ce-atl-1 appears to be important for early embryogenesis, and loss of its function results in a defect in chromosome segregation, similar to what has been observed for AT related proteins. PMID- 11016842 TI - The construction of the first balancer chromosome for the Mediterranean fruit fly, Ceratitis capitata. AB - The construction of the first balancer chromosome, FiM1, for the medfly Ceratitis capitata is described. This chromosome has three overlapping pericentric inversions and is marked with dominant and recessive mutations. The inversion breakpoints of FiM1 suppress recombination throughout the length of the fifth chromosome, allowing lethal mutations to be recovered and maintained. This chromosome will provide a powerful tool for the manipulation of laboratory stocks, in particular, the recovery of new mutant and transgenic strains. We demonstrate the use of FiM1 for the recovery and maintenance of chromosomes carrying lethal mutations. PMID- 11016843 TI - Critical base substitutions that affect the splicing and/or homing activities of the group I intron bi2 of yeast mitochondria. AB - The second intron (bi2) of the cyt b gene from related Saccharomyces species has an extraordinarily conserved sequence and can have different functions in wild type cells. The protein encoded by the S. cerevisiae intron functions as a maturase to promote intron splicing, while the homologous S. capensis intron encodes a bifunctional protein that acts both as a maturase and as a homing endonuclease (I-ScaI) promoting intron mobility. The protein encoded by intron bi2 belongs to a large gene family characterized by the presence of two conserved LAGLIDADG motifs (P1 and P2). In this study, we analysed a set of splicing deficient mutants of the S. cerevisiae intron bi2 that carry non-directed mutations affecting the maturase activity, and a set of directed missense mutations introduced into the bifunctional protein encoded by the S. capensis intron. Analysis of these mutations has allowed identification of the residues in the conserved P1 and P2 motifs which are crucial for splicing and homing activities. Moreover, several mutations which are located in the C-terminal part of the protein have been found to affect both functions. PMID- 11016844 TI - Expression of the barley dehydrin multigene family and the development of freezing tolerance. AB - Dehydrins (DHNs; LEA D11) are one of the typical families of plant proteins that accumulate in response to dehydration, low temperature, osmotic stress or treatment with abscisic acid (ABA), or during seed maturation. We previously found that three genes encoding low-molecular-weight DHNs (Dhn1, Dhn2 and Dhn9) map within a 15-cM region of barley chromosome 5H that overlaps a QTL for winterhardiness, while other Dhn genes encoding low- and high-molecular-weight DHNs are located on chromosomes 3H, 4H and 6H. Here we examine the expression of specific Dhn genes under conditions associated with expression of the winterhardiness phenotype. Plants grown at 4 degrees C or in the field in Riverside, California developed similar, modest levels of freezing tolerance, coinciding with little low-MW Dhn gene activity. Dicktoo (the more tolerant cultivar) and Morex (the less tolerant) grown in Saskatoon, Canada expressed higher levels of expression of genes for low-MW DHNs than did the same cultivars in Riverside, with expression being higher in Dicktoo than Morex. Dehydration or freeze-thaw also evoked expression of genes for low MW DHNs, suggesting that the dehydration component of freeze-thaw in the field induces low expression of genes encoding low-MW DHNs. These observations are consistent with the hypothesis that the major chilling-induced DHNs help to prime plant cells for acclimation to more intense cold, which then involves adaptation to dehydration during freeze-thaw cycling. A role for chromosome 5H-encoded DHNs in acclimation to more intense cold seems possible, even though it is not the basis of the major heritable variation in winterhardiness within the Dicktoo x Morex population. PMID- 11016845 TI - Characterization of the Neurospora crassa mus-25 mutant: the gene encodes a protein which is homologous to the Saccharomyces cerevisiae Rad54 protein. AB - Characterization of the Neurospora crassa mus-25 mutant suggests that it is defective in recombination repair and belongs to the uvs-6 epistasis group. It shows a high sensitivity to the alkylating agents methyl methanesulfonate (MMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), but not to UV radiation. It is barren (i.e. does not produce ascospores) in homozygous crosses. The frequency of MMS-induced mutations at the ad-3 loci is approximately three times higher than in the wild type. The ratio of homologous to nonhomologous integration of the pMTR::HYG plasmid is much lower than in wild type. The mus-25 mutant is epistatic to the mei-3 mutant for MMS sensitivity. mei-3, which is a homololog of the Saccharomyces cerevisiae gene RAD51, is a member of the uvs-6 epistasis group which contains several genes that are homologous to recombination repair genes in other organisms. The mus-25 gene was cloned by identifying a genomic DNA fragment which complements the MMS sensitivity of the mutant. The amino acid sequence deduced from the cloned DNA showed a high degree of homology to the Rad54 protein, which is involved in recombinational repair in S. cerevisiae. Comparison of the nucleotide sequences of the genomic and cDNAs of the mus-25 gene revealed an ORF of 2505 bp with a single 118-bp intron beginning immediately after the second nucleotide of the AUG start codon. The molecular weight of the deduced gene product was 93.5 kDa. The transcript level was raised within 60 min after UV irradiation or MMS treatment, as also observed for the expression of the other N. crassa recombinational repair genes, suggesting the existence of a common mechanism which induces expression of the recombinational repair genes in response to DNA damage. PMID- 11016846 TI - A gene cluster from Streptomyces galilaeus involved in glycosylation of aclarubicin. AB - We have cloned and characterized a gene cluster for anthracycline biosynthesis from Streptomyces galilaeus. This cluster, 15-kb long, includes eight genes involved in the deoxyhexose biosynthesis pathway, a gene for a glycosyltransferase and one for an activator, as well as two genes involved in aglycone biosynthesis. Gene disruption targeted to the activator gene blocked production of aclacinomycins in S. galilaeus. Plasmid pSgs4, containing genes for a glycosyltransferase (aknS), an aminomethylase (aknX), a glucose-1-phosphate thymidylyltransferase (akn Y) and two genes for unidentified glycosylation functions (aknT and aknV), restored the production of aclacinomycins in the S. galilaeus mutants H063, which accumulates aklavinone, and H054, which produces aklavinone with rhodinose and deoxyfucose residues. Furthermore, pSgs4 directed the production of L-rhamnosyl-epsilon-rhodomycinone and L-daunosaminyl-epsilon rhodomycinone in S. peucetius strains that produce epsilon-rhodomycinone endogenously. Subcloning of the gene cluster was carried out in order to further define the genes that are responsible for complementation and hybrid anthracycline generation. PMID- 11016847 TI - Schizosaccharomyces pombe ehs1p is involved in maintaining cell wall integrity and in calcium uptake. AB - The Schizosaccharomyces pombe mutant ehs1-1 mutant was isolated on the basis of its hypersensitivity to Echinocandin and Calcofluor White, which inhibit cell wall synthesis. The mutant shows a thermosensitive growth phenotype that is suppressed in the presence of an osmotic stabiliser. The mutant also showed other cell wall-associated phenotypes, such as enhanced sensitivity to enzymatic cell wall degradation and an imbalance in polysaccharide synthesis. The ehs1 + gene encodes a predicted integral membrane protein that is 30% identical to Saccharomyces cerevisiae Mid1p, a protein that has been proposed to form part of a calcium channel. As expected for such a function, we found that ehs1+ is involved in intracellular Ca2+ accumulation. High external Ca2+ concentrations suppressed all phenotypes associated with the ehs1 null mutation, suggesting that the cell integrity defects of ehs1 mutants result from inadequate levels of calcium in the cell. We observed a genetic relationship between ehs1+ and the protein kinase C homologue pck2+. pck2+ suppressed all phenotypes of ehs1-1 mutant cells. Overproduction of pck2p is deleterious to wild-type cells, increasing 1,3-beta-D-glucan synthase activity and promoting accumulation of extremely high levels of Ca2+. The lethality associated with pck2p, the increase in 1,3-beta-D-glucan synthase production and the strong Ca2+ accumulation are all dependent on the presence of ehs1p. Our results suggest that in fission yeast ehs1p forms part of a calcium channel that is involved in the cell wall integrity pathway that includes the kinase pck2p. PMID- 11016848 TI - Regulation of septation: a novel role for SerC/PdxF in Salmonella? AB - The sfiW locus of Salmonella enterica, previously identified by mutations that suppress the cell division defect of His-constitutive (His(c)) strains, corresponds to serC, the bifunctional gene for phosphoserine-oxoglutarate aminotransferase (SerC) and 2-ketoerythroic acid 4-phosphate transaminase (PdxF). SerC- mutants form small, nearly spherical cells in a wild-type (His+) background, suggesting that the SerC/PdxF product acts as a septation antagonist. Suppression of His(c) filamentation by serC mutations may be explained by loss of the anti-septation activity of SerC/PdxF. The isolation of serC alleles that have lost their biosynthetic activities but are still able to inhibit septum formation suggests that the anti-septation activity of the SerC/PdxF product is unrelated to its known roles in serine and pyridoxine biosynthesis. PMID- 11016849 TI - Differences in regulation of yeast gluconeogenesis revealed by Cat8p-independent activation of PCK1 and FBP1 genes in Kluyveromyces lactis. AB - The yeast Kluyveromyces lactis is can utilise a wide range of non-fermentable carbon compounds as sole sources of carbon and energy, and differs from Saccharomyces cerevisiae in being able to carry out oxidative and fermentative metabolism simultaneously. In S. cerevisiae, growth on all non-fermentable carbon sources requires Cat8p, a transcriptional activator that controls the expression of gluconeogenic and glyoxylate cycle genes via CSREs (Carbon Source Responsive Elements). The down-regulation of Cat8p by fermentable carbon sources is the primary factor responsible for the tight repression of gluconeogenesis by glucose in S. cerevisiae. To analyse the regulation of gluconeogenesis in K. lactis, we have cloned and characterised the K. lactis homologue of CAT8 (KlCAT8). The gene was isolated by multicopy suppression of a fog2/klsnf1 mutation, indicating a similar epistatic relationship between KlSNF1 and KlCAT8 as in the case of the S. cerevisiae homologues. KlCAT8 encodes a protein of 1445 amino acids that is 40% identical to ScCat8p. The most highly conserved block is the putative Zn(II)2Cys6 DNA-binding domain, but additional conserved regions shared with members of the zinc-cluster family from Aspergillus define a subfamily of Cat8p-related proteins. KlCAT8 complements the growth defect of a Sccat8 mutant on non fermentable carbon sources. In K. lactis, deletion of KlCAT8 severely impairs growth on ethanol, acetate and lactate, but not on glycerol. Derepression of enzymes of the glyoxylate cycle--malate synthase and particularly isocitrate lyase--was impaired in a Klcat8 mutant, whereas Northern analysis revealed that derepression of KlFBP1 and KlPCK1 does not require KlCat8p. Taken together, our results indicate that in K. lactis gluconeogenesis is not co-regulated with the glyoxylate cycle, and only the latter is controlled by KlCat8p. PMID- 11016850 TI - Cloning and characterization of a Legionella pneumophila-specific gene encoding a member of the LysR family of transcriptional regulators. AB - Flagellin gene regulation in Legionella pneumophila is modulated by various environmental factors. The expression of the virulent phenotype seems to be linked genetically to flagellum expression. To better understand the mechanisms of flagellin gene expression in L. pneumophila (Lp), we screened a pool of plasmids from a L. pneumophila Corby genomic library for the ability to prevent or reduce luciferase activity in the Escherichia coli strain YK410, which harbours a Lp-pflaA-luxAB fusion. We cloned a DNA fragment encoding the N terminal part of a protein with significant similarity to members of the LysR family of transcriptional regulators (LTTRs). The entire gene, cloned by inverse PCR, was named flaR. It encodes a protein of 302 amino acids, and computer assisted analysis of the amino acid sequence revealed a helix-turn-helix motif located near the N-terminus of the protein. The FlaR protein exhibits 21-31% identity to various LTTRs. Furthermore, gel retardation experiments indicate that the FlaR protein is able to bind to its own promoter region and, to a lesser extent, to the flaA promoter of L. pneumophila. The flaR promoter region contains putative LysR binding motifs and two putative Fur boxes. Taken together, these results indicate that FlaR is a DNA-binding protein which belongs to the LTTR family. Southern analysis with a L. pneumophila Corby-specific flaR probe revealed homologous genes in various L. pneumophila strains, but not in the 12 nonpneumophila strains tested so far. PMID- 11016851 TI - Dendritic cell activation by danger and antigen-specific T-cell signalling. AB - Recent transplantation, animal and in vitro studies suggest a dependence of some immune reactions on tissue damage. Although many factors involved in enhancing immune responses through tissue damage have yet to be identified, recent data suggests that one of the targets of these cellular stress factors is the bone marrow derived dendritic cell (DC). DC are potent initiators of primary immune responses and hold the key to immune reactions through their ability to sense changes in their local environment and respond appropriately to induce T-cell immunity, or possibly tolerance. In the lymph node, DC are also influenced by antigen-specific signalling from T cells, which may extend and amplify DC antigen presenting capabilities, especially for the stimulation of cytotoxic responses. It now appears that both tissue damage and antigen-specific T-cell derived signals act together on the DC to promote the appropriate immune reaction to antigen. Thus DC antigen presenting behaviour is not only dependent on the context of antigen encounter in the periphery, but also on the availability of antigen-specific T cells and their T-cell receptor specificities. PMID- 11016852 TI - Genomic organization and amplification of the human plakoglobin gene (JUP). AB - Plakoglobin is a globular protein common to the intracellular plaques of adhesive junctions, predominantly desmosomes and adherens junctions. Recently, a number of pathogenic mutations have been described in other components of desmosomes, specifically in plakophilin 1, desmoplakin and desmoglein 1. The phenotype of affected patients mainly involves thickening of palm and sole skin (keratoderma). Although no human mutations in plakoglobin have been described thus far, this protein represents an excellent candidate for other human genetic disorders, possibly involving skin and heart, sites of high plakoglobin expression. To facilitate future mutation detection analyses in such conditions, we have characterized the intron-exon organization of the human plakoglobin gene, which comprises 13 distinct exons spanning approximately 17 kb on 17q21. We have also developed a PCR-based mutation detection strategy using primers placed on flanking introns followed by direct sequencing of the PCR products. PMID- 11016853 TI - VEGF production, cell proliferation and apoptosis of human IGR 1 melanoma cells under nIFN-alpha/beta and rIFN-gamma treatment. AB - The effect of natural and recombinant interferons (nIFN, rIFN) on cell growth, apoptosis and the production of vascular endothelial growth factor (VEGF) was investigated in the human melanoma cell line IGR 1. We determined cell proliferation, cell vitality, DNA synthesis, apoptosis, intracellular oxygen radicals (ROS) and VEGF-mRNA as well as VEGF-protein levels. rIFN-gamma significantly inhibited growth by decreasing DNA synthesis and increasing apoptosis. Less pronounced was the growth inhibitory effect of nIFN-beta because an increased rate of apoptosis was outweighed by enhanced DNA synthesis. nIFN alpha only had minor effects on cell growth parameters. Under long-term incubation (144 h) nIFN-beta decreased, but rIFN-gamma increased production of the angiogen VEGF. Our data underscore the multiple effects of IFNs on melanoma cells and may contribute to the understanding of ambivalent results of melanoma therapy by IFNs. Particularly, the increased VEGF production under long-term treatment with serum IFN levels between 100 and 1,200 IU/ml should be kept in mind. PMID- 11016854 TI - Circulating levels of soluble E-selectin, ICAM-1 and VCAM-1 in bullous pemphigoid during low-dose methotrexate therapy. A prospective study. AB - Soluble iso-forms of cellular adhesion molecules (sCAMs) have been described and reported to be elevated in various inflammatory diseases. Elevated levels of sE selectin have recently been detected and found to correlate with the number of blisters in bullous pemphigoid (BP) during oral corticosteroid therapy. In this prospective study we analysed levels of sCAMs in 10 elderly BP patients during low-dose oral pulse methotrexate monotherapy. We used standardised ELISA kits for soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1) and sE-selectin on 65 sera from 10 patients and 19 controls. Results were correlated with clinical parameters. Before therapy, we found significant elevation of sE-selectin (P=0.004) and sVCAM-1 (P=0.002) but not of sICAM-1. sE-selectin levels decreased during the efficient therapy and correlated with the number of blisters. Our results further support the proposition that sE-selectin might be a future clinical and predictive tool; but whether the elevation of sVCAM-1 also might reflect the disease activity in BP needs more investigation. The findings also indicate that BP might be more a cellularly mediated disease where interactions of different adhesion molecules play a crucial role. PMID- 11016855 TI - Cytokine expression patterns distinguish HIV associated skin diseases. AB - AIDS is known to cause a shift of cytokines in the periphery. However, predominant cytokines in skin of patients with HIV-associated skin diseases have not been clearly defined. We hypothesized that there are distinct cytokine profiles that distinguish among the different clinical manifestations of AIDS related skin diseases. To test this hypothesis, lesional and non-lesional skin was biopsied from 53 HIV+ patients with Kaposi's sarcoma (KS), psoriasis, and pruritus due to eosinophilic folliculitis, and from HIV negative controls with psoriasis or KS prior to therapy. Immunohistochemistry was performed with antibodies to tumor necrosis factor (TNF)-alpha, interleukin (IL)-10, interferon (IFN)-gamma, and interferon-inducible protein (IP)-10. HIV positive individuals included 10 with psoriasis, 14 with pruritus, and 15 with Kaposi's sarcoma. HIV negative controls included 12 with psoriasis and two with KS. Semi-quantitative analysis of cytokine staining was confirmed by optical density using a digital imaging system on four representative skin sections from each disease. Optical density analyses were conducted using ANOVA and t-tests. We found that epidermis overlying HIV+ Kaposi's sarcoma was hyperproliferative and was highest in IP-10, IFN-gamma, and IL-10 (P=0.0001). HIV+ pruritus was significantly highest in TNF alpha (P=0.0001) staining. HIV+ psoriasis represented an intermediate state for all four cytokines. Normal skin adjacent to lesions showed the same relative patterns, with lower intensities. Skin diseases seen frequently in the setting of HIV and immunodeficiency have relatively distinct levels and patterns of cytokine expression that may reflect immune dysfunction, reactivity to HIV and to opportunistic infections. PMID- 11016856 TI - Therapeutic interventions in mice with chronic proliferative dermatitis (cpdm/cpdm). AB - Chronic proliferative dermatitis (cpd) is a spontaneous mutation in C57BL/Ka mice (cpdm/cpdm). The dermatitis is characterized by redness, hairloss, scaling, pruritus and histologically by epithelial hyperproliferation, infiltration of eosinophils, macrophages and mast cells. Lesions similar to those in the skin occur in the esophagus and forestomach. In this paper, we describe the effect of drug treatments directed against epidermal hyperproliferation (calcipotriene and etretinate), against inflammation (corticosteroids and dapsone) and against pruritus (loratidine and capsaicin). The criteria used to objectively estimate the effect of the treatment were 1) macroscopic evaluation of the lesions (cpd score), 2) degree of epithelial hyperproliferation assessed by BrdU incorporation and epithelial thickness, and 3) microscopic evaluation of the inflammatory cells in the skin samples. Treatment of the cpdm/cpdm mice with calcipotriene (5 microg/day for 3 weeks) inhibited epidermal proliferation and the number of eosinophils. Systemic etretinate treatment (30 microg/g/day for 3 weeks) was not very effective. Topical corticosteroids (0.05 microg/day, for 3 weeks) exerted a therapeutic effect on the hyperproliferation and the number of eosinophils. Oral dapsone treatment (34 microg/g/day, for 5 weeks) reduced the BrdU incorporation in the skin and the epithelial thickness in the esophagus. The anti-histamine loratidine (orally, 1.7 microg/ g/day, for 4 weeks) reduced the severity of the lesions macroscopically, probably by suppressing the pruritus. Capsaicin (topically, 30 mM, for 5 weeks) also reduced the severity of the macroscopic observable lesions. Moreover, capsaicin reduced the dorsal and ventral epidermal thickness. The results from this and previous studies indicate that steroids (topically and systemically) and less strongly calcipotriene are the most effective treatments for the lesions observed in the cpdm/cpdm mice, since both hyperproliferation and the influx of eosinophils are reduced. Although the pathogenesis of the cpd lesions remains to be determined, our results indicate that the cpdm/cpdm mouse can be used to investigate new drugs for their possible application in chronic dermatitis. PMID- 11016857 TI - Is psoriasis a T-cell disease? AB - The etiology and pathogenesis of psoriasis--one of the most common chronic, inflammatory, hyperproliferative skin disorders of man--have long fascinated dermatologists, pathologists and biologists alike. Here, we have a model disease that offers to study neuroectodermal-mesenchymal interactions in the widest sense possible. Epithelial, endothelial, and hematopoietic cells as well as neurons projecting into the skin apparently all interact with each other to generate the characteristic psoriatic lesion. For decades, the ongoing controversy on the molecular nature, choreography and hierarchy of these complex interactions e.g. between epidermal keratinocytes, T cells, neurotrophils, endothelial cells and sensory nerves has served as a driving force propelling investigative dermatology to ever new horizons. This debate has not only been at the heart of our quest to develop more effective forms of therapy for this socially crippling disease, but it also has profoundly influenced how we view the skin as a whole: the numerous competing theories on the pathogenesis of psoriasis published so far also are reflections on the evolution of mainstream thought in skin biology over the last decades. These days, conventional wisdom infatuated with a T-cell-centered approach to inflammatory skin diseases-- portrays psoriasis as an autoimmune disease, where misguided T lymphocyte activities cause secondary epithelial abnormalities. And yet, as this CONTROVERSIES feature reminds us, some authoritative "pockets of academic resistance" are still quite alive, and interpret psoriasis e.g. as a genetically determined, abnormal epithelial response pattern to infectious and/or physicochemical skin insults. Weighing the corresponding lines of argumentation is not only an intriguing, clinically relevant intellectual exercise, but also serves as a wonderful instrument for questioning our own views of the skin universe and its patterns of deviation from a state of homeostasis. PMID- 11016858 TI - Excitation-contraction coupling in cardiac muscle revisited. AB - According to the current views the direct and indispensable source of Ca2+ activating contraction is sarcoplasmic reticulum (SR). Ca2+ is released from the SR when its release channels (ryanodine receptors) are activated by Ca2+ influx through the L-type Ca2+ channels (dihydropyridine receptors). In contrast, ryanodine receptors of skeletal muscles are activated by conformational changes in dihydropyridine receptors induced by sarcolemmal voltage. Ca2+ influx is not necessary for their activation. In this review the papers not quite conforming with the current views are referred to and discussed. Their results suggest that SR is not an indispensable source of contractile Ca2+ at least in some mammalian species, and that cardiac ryanodine receptors may be activated by conformational changes in dihydropyridine receptors without Ca2+ influx (like in skeletal muscle). This may be a mechanism parallel to or accessory to the Ca2+ induced release of Ca2+ (CIRC). PMID- 11016859 TI - Differential responses of hematopoietic and non-hematopoietic cells to anti inflammatory cytokines: IL-4, IL-13 and IL-10. AB - Recombinant preparations of human anti-inflammatory cytokines: IL-4, IL-13 and IL 10, inhibited LPS-induced synthesis of TNFalpha and IL-6 in the whole human blood tested in vitro. These cytokines also inhibited LPS-induced IL-6 and TNF mRNA accumulation in isolated human blood monocytes/macrophages. On the other hand, similar concentrations of IL-4 and IL-13 (but not IL-10) enhanced synthesis of IL 6 in cultured human umbilical vein endothelial cells (HUVEC). In human hepatoma HepG2 cells IL-4 and IL-13 (but not IL-10) inhibited IL-6-induced synthesis of haptoglobin. These differential responses to the tested anti-inflammatory cytokines were observed at mRNA and protein levels and may reflect cell specificities in signalling pathways and gene expression. When HUVEC and HepG2 cells were cultured together and stimulated with LPS the addition of IL-4 or IL 13 resulted in the reduction of LPS-induced and IL-6-mediated haptoglobin synthesis. Thus in co-culture the inhibitory effects of IL-4 or IL-13 on HepG2 cells prevail over stimulation of IL-6 synthesis in HUVEC. PMID- 11016860 TI - The role of dopamine D2 receptor in the behavioral effects of imipramine--study with the use of antisense oligonucleotides. AB - Antisense strategies have a potential to specifically block the production of a given protein, e.g. receptor subtype, thus may help to uncover its behavioral and/or biochemical function. In the present study we demonstrated the utility of this approach for studying the role of dopamine D2 receptors in the anti immobility effect of imipramine in the forced swimming test. Following intracerebroventricular (i.c.v.) administration of phosphorothioate oligonucleotide complementary to mRNA encoding for dopamine D2 receptors (D2 antisense ODN; 1 nmol/1 microl H2O, twice a day for 5 days) to the rats, the decrease in the locomotor activity (shortened total distance travelled and decrease in vertical activity, without differences in the stereotypic movements of animals), as well as the decrease of specific binding of [3H]raclopride in the striatum and limbic forebrain were observed. At the same time, i.c.v. administration of D2 antisense ODN reversed the effect of imipramine in the forced swimming test, what may indicate that the dopamine D2 receptors play a significant role in the behavioral anti-immobility effects of imipramine. PMID- 11016861 TI - Endogenous nitric oxide proves not to be involved in the inhibition by IL-1beta of TGF-alpha-stimulated proliferation of RGM1 cells. AB - Combined treatment of isolated rat gastric mucosal (RGM1) cells with interleukin (IL)-1beta and transforming growth factor (TGF)-alpha resulted in expression of inducible nitric oxide synthase (iNOS) mRNA and iNOS protein 24 hr after the treatment. Constitutive NOS (nNOS) protein was not proved in the cells and not activated by IL-1beta+TGFalpha. Although IL-1beta and TGF-alpha alone exerted little or no effect on NO2 production, their combination gradually increased NO2- production from 12 to 24 hr following treatment. NO2- production stimulated by IL 1beta + TGFalpha was significantly reduced by N(G)-nitro-L-arginine methyl ester (L-NAME) or aminoguanidine, yet not by D-NAME. S-nitroso-N-acetyl-D,L penicillamine and sodium nitropruside significantly inhibited both spontaneous and TGF-alpha stimulated DNA synthesis. Nonetheless, L-NAME did not affect the inhibition by IL-1beta of TGF-alpha-stimulated proliferation of RGM1 cells, eliminating the possibility of involvement of NO in the underlying mechanisms. PMID- 11016862 TI - Omeprazole fails to suppress up-regulation of gastric mucosal endothelin converting enzyme-1 by Helicobacter pylori lipopolysaccharide. AB - BACKGROUND: Endothelin-1, a key mediator of inflammatory processes, is produced from its biologically inactive precursor, big ET- by the action of endothelin converting enzyme-1(ECE-1). In this study, we applied the animal model of H. pylori lipopolysaccharide-induced gastritis to assess the effect of three different types of antiulcer agents on the gastric mucosal expression of ECE-1 activity. METHODS: Rats, pretreated twice daily for 3 days with proton pump inhibitor, omeprazole at 40 mg/kg, gastroprotective agent, sulglycotide at 200 mg/kg, H2-receptor antagonist, ebrotidine at 100 mg/kg or the vehicle, were subjected to intragastric application of H. pylori lipopolysaccharide at 50 microg/animal, and after 2, and 4 additional days on the drug or vehicle regimen their mucosal tissue used for histologic and biochemical assessment. RESULTS: In the absence of antiulcer agents, H. pylori lipopolysaccharide elicited a pattern of mucosal inflammatory responses resembling that of acute gastritis which reached a maximum by the 4th day and were accompanied by a 4.1-fold increase in the mucosal expression of ECE-1 activity and an 8.8-fold enhancement in TNF alpha. Treatment with sulglycotide led to a 56.7% reduction in the extent of mucosal inflammatory involvement, the mucosal expression of ECE-1 activity fell by a 40.5% and the level of TNF-alpha declined by a 69%. Ebrotidine produced a 50.9% decrease in the extent of mucosal inflammatory involvement, a 33.6% decrease in the expression of ECE-1 activity and a 64.1% decline in TNF-alpha, whereas omeprazole elicited a 37.6% reduction in the extent of mucosal inflammatory involvement and a 29.5% decrease in TNF-alpha, but had no effect on the lipoploysaccharide-induced increase in the mucosal expression of ECE-1 activity. CONCLUSIONS: The findings implicate up-regulation of ECE-1 in triggering the induction of TNF-alpha and propagation of gastric mucosal inflammatory responses to H. pylori. We also show that omeprazole, in contrast to sulglycotide and ebrotidine, fails to counter the enhancement in the mucosal expression of ECE-1 caused by H. pylori- lipopolysaccharide. PMID- 11016863 TI - Impact of ethanol on innate protection of gastric mucosal epithelial surfaces and the risk of injury. AB - Earlier investigations on the effect of ethanol on synthesis and posttranlational glycosylation of gastric mucus glycoprotein (mucin) revealed quantitative changes in the apoprotein assembly, glycosylation, and mucin retention on the mucosal surface (Slomiany et al.., Alcoholism: Clin. Exp. Res. 21, 417-423, 1998). To assess whether metabolic consequences of ethanol ingestion, documented in the in vitro system are also occurring in vivo the rats were subjected to 8 weeks of ethanol containing liquid diet. The retention of mucin on the surface of gastric mucosa was quantitated by measuring the binding of gastric mucin to Mucin Binding Protein (MBP) of gastric mucosa. The results were compared with those obtained with the rats subjected to pair-feeding the isocaloric-control diet. Before alcohol administration, and in two weeks' intervals thereafter, the gastric contents from the animals was collected and mucin purified. After 8 weeks of the respective diet, the animals were sacrificed and their gastric mucosa used for MBP preparation. The binding of mucin to MBP before ethanol, and after 2, 4, 6, and 8 weeks of ethanol diet was quantitated with Enzyme Linked Lectin Assay (ELLA). The study with standard mucin revealed that binding of mucin to MBP differs substantially between individual animals. The same variability in binding was observed with the individual mucin preparations collected at the onset of the experiment. However, with the progression of ethanol feeding, the mucin samples besides displaying the variable and animal-specific binding to MBP at the initiation of the experiment, also showed a dramatic decrease in binding. In five animals, after two weeks of ethanol diet, mucin binding to MBP decreased by 50%; in two animals, the drastic decrease in binding was observed in mucin collected after four weeks of alcohol feeding; and in one animal a 20% decrease in binding persisted for six weeks, and then decreased to 50% in the last collection. Also, in two animals, the mucin collected after 8 weeks of ethanol feeding retained only 6-9% of the initial binding capacity. In contrast, in pair-fed controls, the mucin binding to MBP remained the same or increased up to 20%. Results of the studies, performed on mucin of the individual animals and matching preparations of MBP, showed that each animal expresses different degree of mucin binding. Moreover, in chronic ethanol ingestion, the individual variations are accompanied by a decrease in mucin binding to MBP. Since the observed decrease in binding occurred in samples containing the same preparation of MBP, the component affected by alcohol resides on mucin. Thus, considering the in vitro impact of ethanol on generation of carbohydrate chains in Golgi, and the finding on mucin oligosaccharides-dependent mucin-MBP complex formation, we conclude that ethanol impairs the synthesis of mucin oligosaccharide structures required for binding with MBP, and the retention on gastric mucosal surfaces. PMID- 11016864 TI - The influence of sensory nerves and CGRP on the pancreatic regeneration after repeated episodes of acute pancreatitis in rats. AB - Stimulation of capsaicin sensitive nerves or administration of calcitonin gene related peptide (CGRP) before induction of acute pancreatitis (AP) attenuates pancreatic damage, whereas CGRP administration after development of AP aggravates lesion of pancreatic tissue. The aim of this study was to determine the effect of prolonged activity of sensory nerves or CGRP administration on the pancreatic repair after repeated episodes of AP. Five episodes of acute caerulein-induced pancreatitis (10 microg/kg/h for 5 h s.c.) were performed at weekly intervals in rats receiving either vehicle or capsaicin at the sensory nerve stimulatory dose (0.5 mg/kg, 3 times daily), or CGRP (10 microg/kg, 3 times daily). Two weeks after the last induction of AP morphological signs of pancreatic damage, pancreatic blood flow (PBF), serum and pancreatic amylase activity, fecal chymotrypsin activity, pancreatic weight, pancreatic RNA and DNA content, as well as, serum interleukin-1beta (Il-1beta ) were assessed. Pancreata of animals receiving vehicle alone showed almost full recovery within two weeks after last episode of pancreatitis induction. In capsaicin-treated group of rats, we observed the increase in PBF by 44% and in serum Il-1beta concentration by 91%. The pancreatic amylase activity, fecal activity of chymotrypsin, pancreatic nucleic acids content and DNA synthesis were decreased. In rats treated with CGRP the alterations in PBF, serum Il-1beta concentration, as well as, in pancreatic and fecal activity of enzymes were similar to capsaicin treated group but less pronounced. We conclude that prolonged activity of capsaicin-sensitive sensory nerves and the presence of their main mediator-CGRP during pancreatic regeneration after AP leads to pancreatic functional insufficiency typical for chronic pancreatitis. PMID- 11016865 TI - Temporary elevation of pancreatic lysosomal enzymes, as a result of the omeprazole-induced peripancreatic inflammation in male Wistar rats. AB - Omeprazole is one of the substituted benzimidazoles, which is not free of side effects. The aim of the study was to evaluate the influence of omeprazole therapy on pancreas. Omeprazole was administered intraperitoneally, twice a day, for 3 days to the male rats in 0.571 mg/kg b.w. and 5.71 mg/kg b.w. doses. Half of animals were sacrificed in the 4th day of the experiment. The remaining rats were raised for another 6 weeks, without any xenobiotics, and sacrificed on the 47th day. The activity of acid phosphatase, beta-galactosidase, cathepsin B, and L, lipase, N-acetyl-glucosaminidase, and sulphatase was evaluated. The slides of the pancreas were examined in light microcopy in hematoxylin-eosin, asan, periodic acid-Schiff (paS) stains. Statistical increase in total activities of acid phosphatase, beta-galactosidase, lipase, N-acetyl-glucosaminidase, sulphatase, and acute inflammatory infiltration in peripancreatic fat tissue without histological pancreas impairment, were observed after the higher dose on the 4th day of experiment. Histological picture and enzymatic profiles were normalized during the next 6 weeks. We concluded that intraperitoneal administration of omeprazole causes tissue inflammation in the peripancreatic lipid tissue and reactive elevation of some pancreatic lysosomal enzymes. PMID- 11016866 TI - The effect of naloxone on trigemino-hypoglossal reflex inhibited by periaqueductal central gray stimulation in rats. AB - The aim of the study was to determine whether opioid receptor antagonist naloxone abolishes the influence of periaqueductal central gray (PAG) on nociceptive evoked tongue jerks (ETJ) - a trigemino-hypoglossal reflex induced by tooth pulp stimulation. In rats under chloralose anesthesia three subsequent series of perfusions of lateral ventricles - cerebellomedullary cistern with Mc Ilwain Rodnight's solution, Met-enkaphalin (Enk-Met) and naloxone were carried out. The amplitudes of tongue jerks induced by tooth pulp stimulation were recorded during subsequent 10 min perfusions. Mean amplitude of tongue movements induced by tooth pulp stimulation was regarded as the indicator of the magnitude of trigemino hypoglossal reflex. We observed that perfusion of the cerebral ventricles with Enk-Met (100 nmol/mL) inhibited the trigemino-hypoglossal reflex by 46%, whereas naloxone (100 nmol/mL), added to the solution perfusing the cerebral ventricles system, increased the reflex by 42%. The amplitude of ETJ was significantly reduced during PAG stimulation with a train of electrical impulses. After obtaining a significant 93% - inhibition of ETJ, naloxone (100 nmol/mL) was added to the perfusion fluid. This led to a significant increase of the reflex by 68%. The above results suggest that the inhibition of ETJ due to PAG stimulation is partially reversed by naloxone and mediated via interactions with endogenous opioid systems involved in modulation of nociception. PMID- 11016867 TI - Functional studies on sciatic nerve blood flow in respect to its vascular supply and tonic neural activity. AB - An introduction of laser flow meters for a continuous measurements of a tissue blood flow has opened new avenues for an accurate assessment blood flow in peripheral nerves. The aim of our study was: 1) to carry out a functional verification of anatomical sources of a sciatic nerve blood supply in the rat; 2) develop a measurement technique to facilitate standardisation of results; 3) to determine the role of nerve fibres tonic activity in the maintenance of a resting blood flow in the sciatic nerve. Based on results of the present study the following conclusions have been drawn out: 1) in order to obtain a real values of the blood flow through the sciatic nerve it is necessary to remove its muscular fascia; 2) an uninjured epineurium plays a crucial role in maintaining the resting blood flow; 3) major blood supply of sciatic nerve comes from inferior gluteal and popliteal arteries; 4) the tonic neural activity plays a role in the maintenance of the resting sciatic nerve blood flow in anaesthetised rats. PMID- 11016868 TI - Bidirectional action of extracellular ATP on intracapillary volume of isolated rat renal glomeruli. AB - Receptors for extracellular nucleotides (P2-purinoceptors) are expressed in renal glomerulus; both on mesangial and endothelial cells. In the present study we have evaluated the potential role of ATP in the regulation of glomerular contraction and relaxation. Using [3H]-inulin we measured the Glomerular Inulin Space (GIS), (that reflects mainly glomerular intracapillary volume), in the presence of ATP and its analogues e.g. 2-methylthio-ATP (P2Y-receptor agonist) and beta,gamma methylene-ATP (P2X-receptor agonist). Incubation of the intact glomeruli with ATP or 2-methylthio-ATP or beta,gamma-methylene-ATP induced a decrease of GIS in similar magnitude as angiotensin II e.g.: about 10% of the basal value. When glomeruli were precontracted with angiotensin II it was observed that both ATP and 2-methylthio-ATP induced an increase of GIS to the basal value, similarly to atrial natriuretic factor. Furthermore, there was no relaxing effect with beta,gamma-methylene-ATP. We suggest that, these bidirectional changes of the intracapillary volume induced by the extracellular ATP may contribute to regulation of glomerular dynamics. PMID- 11016869 TI - Co-operation between particulate and soluble guanylyl cyclase systems in the rat renal glomeruli. AB - ANP and NO act via different receptors, although inducing the common intracellular messenger - cyclic GMP. However, interaction between both factors remains unclear. Our observations suggested that in the rat kidney glomeruli, activities of the ANP- and NO-dependent guanylyl cyclase systems may be mutually compensated. To check this, we have tested effects of ANP and sodium nitroprusside (SNP) on cGMP synthesis and relaxation of glomeruli contracted with angiotensin II. The glomeruli were isolated from Wistar rats receiving saline (Control), dexamethasone (DEX), deoxycorticosterone (DOCA) or N-c-nitro-L arginine methyl ester (NAME) for 1 or 2 days. In the DEX glomeruli exposed to 100 microM SNP, rate of cGMP synthesis was significantly higher then in the Control (26.3 vs 16.0 pmol/mg.prot./2 min., P<0.05), while 1 microM ANP was markedly less effective (2.8 vs 16.7 pmol/mg.prot./2 min in Control, P<0.01). On the contrary, in NAME group 1 microM ANP stimulated cGMP synthesis up to 35.6 pmol/mg.prot./2 min whereas efficacy of SNP was slightly suppressed. High correlation coefficient (r = 0.979, p<0.01) indicates interrelationship between NO- and ANP-dependent cGMP synthesis. Ability of the glomeruli to relax in response to ANP or SNP was in accord to their ability to cGMP generation. This was confirmed by high correlation (r = 0.845, p<0.001) between degree of relaxation and rate of cGMP synthesis. Our results support strongly the hypothesis that both, ANP and NO dependent systems co-operate in regulation of the function of kidney glomeruli. PMID- 11016870 TI - Antioxidants with carcinostatic activity (resveratrol, vitamin E and selenium) in modulation of blood platelet adhesion. AB - Compounds with potential antiplatelet activity can be used in the therapy of cardiovascular disorders. We investigated the effects of three different antioxidants with carcinostatic property: trans-resveratrol, Trolox a water soluble analog of vitamin E, and inorganic selenocompounds (sodium selenite and selenate) on blood platelet adhesion to fibrinogen (Fg). Adhesion, the initial step of platelet activation, was estimated by the colorimetric method with BCA (bicinchoninic acid) solution in 96-well Fg-coated microtiter dishes. It was shown that resveratrol significantly inhibited adhesion of both thrombin- and ADP activated platelets to Fg. After incubation of platelets for 30 min. at 37 degrees C with resveratrol at the concentration of 100 microg/ml above 40% inhibition of adhesion was achieved. The inhibition of platelet adhesion of Fg caused by Trolox was lower than by resveratrol and at higher concentration (1 mM) reached maximum 12%. We also demonstrated that neither sodium selenite nor selenate significantly altered platelet adhesion to Fg. We conclude that changed adhesion of blood platelets to Fg in the presence of resveratrol and Trolox, but not selenium may be the result of different antioxidative activities of tested compounds. PMID- 11016871 TI - Response of aorta connective tissue matrix to injury caused by vassopressin induced hypertension or hypercholesterolemia. AB - The aim of the study was to investigate the effect of two various atherogenic stimuli (vasopressin-induced hypertension or hypercholesterolemia) on the collagen and glycosaminoglycan (GAG) content in the internal or external part of both thoracic and abdominal aorta, which are differently susceptible to atherosclerosis. Experimental rabbits were divided into four groups: controls, animals injected with physiological saline or vasopressin at the dose of 1 IU/kg from the 1 st to the 25 th day of experiment, respectively. The animals from group 4 were maintained on food, containing 0.25% cholesterol. Only in the vasopressin-treated group, the systolic blood pressure was elevated from 110 mmHg at the beginning, to 166 mmHg at the end of the study. After 14 weeks the aorta was dissected into internal and external parts. GAG fractions were separated and estimated as uronic acids. Collagen was evaluated as the hydroxyproline content in the tissue. Augmented total GAG and heparan sulphate (HS) level, plus no changes in the collagen content were seen in the internal part of the thoracic aorta in rabbits with hypercholesterolemia or hypertension. In the hypertensive animals, the changes were extended to the external part of the aorta and, additionally, comprised the elevation of the chondroitin-4 sulphate (C-4S) content. The two atherogenic stimuli increased the collagen level with no elevation of the GAG content in the abdominal aorta. A convergent effect of the injury, caused by hypertension or hypercholesterolemia on the collagen, total GAG and HS content was shown in the respective parts of the rabbit aortas. The common GAG, increased in the thoracic aorta, stand for the HS, in both hypertensive and hypercholesterolemic rabbits. As the sensitivity to atherosclerosis development in different segments of the aorta varies, they express various responses of the connective tissue matrix to injuries, caused by hypertension or hypercholesterolemia. PMID- 11016872 TI - Effect of triiodothyronine on the content of phospholipids in the rat liver nuclei. AB - The aim of the present study was to examine the effect of triiodothyronine (T3) on the content of phospholipids and on the incorporation of blood-borne palmitic acid into the phospholipid moieties in the nuclei of the rat liver. T3 was administered daily for 7 days, 10 microg x 100 g(-1). The control rats were treated with saline. Each rat received 14C-palmitic acid, intravenously suspended in serum. 30 min after administration of the label, samples of the liver were taken. The nuclei were isolated in sucrose gradient. Phospholipids were extracted from the nuclei fraction and from the liver homogenate. They were separated into the following fractions: sphingomyelin, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine and cardiolipin. The content and radioactivity of each fraction was measured. It was found that treatment with T3 reduced the content of phosphatidylinositol and increased the content of cardiolipin in the nuclear fraction. In the liver homogenate, the content of phosphatidylinositol decreased and the content of phosphatidylethanolamine and cardiolipin increased after treatment with T3. The total content of phospholipids after treatment with T3 remained unchanged, both in the nuclear fraction and in the liver homogenate. T3 reduced the specific activity of phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and cardiolipin and had no effect on the specific activity of sphingomyelin and phosphatidylinositol both in the fraction of the nuclei and the liver homogenate. It is concluded that excess of triiodothyronine affects the content of phospholipids in the nuclei. The changes in the content of phospholipids in the nuclei largely reflect changes in their content in the liver. PMID- 11016873 TI - Free intracellular calcium ([CA2+]i) in opioid sensitive cells of the porcine anterior pituitary. AB - A new experimental model was utilized to study calcium involvement in the mechanism of opioid influence on cultured porcine pituitary cells. The in vitro model involved interactive argon laser cytometry of pituitary cells pre-loaded by three dyes (fluo-3AM, fura-red and naloxone-conjugated to fluorescein). We compared: 1) the kinetics of free intracellular calcium ([Ca2+]i) in anterior pituitary cells of pregnant pigs (day 25-30) treated in vitro with naloxone (NAL) or gonadotrophin-releasing hormone (GnRH) and 2) the distribution of the opioid sensitive cells by image analysis of doubly loaded cells. In experiment 1, the changes in [Ca2+]i of pituitary cells pre-loaded with fluo-3 AM (488(ex)/520(em) nm) in response to NAL (10(-6) M) or to GnRH (10(-8) M) were compared to a control cell group. Repetitive line scans across cells were performed and the fluorescence emission from individually selected cells was measured in a time dependent manner (in 0.5 seconds intervals during periods of 50 seconds). Analysis of data indicated significant increases of [Ca2+]i in NAL- (P<0.001) and GnRH-treated cells (P<0.05) in comparison to the control group. In experiment 2, the fluorescence intensity of doubly-loaded pituitary cells (fura-red, 488(ex)/605(em) nm, as principal calcium indicator and NAL-conjugated with fluorescein, 488(ex)/520(em) nm, to distinguish opioid-sensitive cells) were measured using dual detector image analysis. We found that only approximately 8% of the entire population of anterior pituitary cells exhibited sensitivity to the opioid antagonist treatment. This paper demonstrates calcium involvement in the opioid action on anterior pituitary cells from pregnant pigs and provides a useful model for studies at the individual pituitary cell level and in time dependent manner. PMID- 11016874 TI - Time dependent and cell-specific action of polychlorinated biphenyls (PCB 153 and PCB 126) on steroid secretion by porcine theca and granulosa cells in mono- and co-culture. AB - To characterize PCB action on follicular cell steroidogenesis two PCB congeners were selected as model substances. PCB 126 because of its dioxin-like configuration and high toxicity and PCB 153 because it is one of the most commonly detected PCB congeners in breast milk. The direct effect of PCBs was investigated using a culture system of porcine theca and granulosa cells collected from porcine preovulatory follicles. Granulosa and theca cells were cultured in M199 medium supplemented with 1, 10 or 100 pg/ml of PCB 126 or 1, 10 and 100 ng/ml of PCB 153. The media were changed after 48, 96 and 144 h and frozen until further estradiol (E2) analysis. Additionally, progesterone (P4) was measured in the granulosa cells culture medium and testosterone (T) in theca cells culture medium. Decrease of testosterone concentration in the theca cells culture medium was found after 96 and 144 hours in culture by both investigated PCB congeners. A decrease in E2 concentration was found after exposure to PCB 153. These findings suggest different actions of two congeners on the steroid synthesis in theca cells. The lack of an increase in E2 secretion after the exposure to PCB 126 could be due to depletion of androgen precursor. In granulosa cell culture PCB153 decreased E2 secretion and increased P4 secretion suggesting luteinization and disruption of aromatization process. PCB 126 in a doses from 1 to 10 pg had no effect on granulosa cells steroidogenesis. However, the highest dose (100 pg) increased concentration of both E2 and P4. This observation suggest that PCB 126 in a pharmacological doses may affect cell membrane permeability, thereby increasing steroid outflow into the medium. These results suggest time dependent and cell-specific differences in PCB 153 and 126 action on follicular cells steroidogenesis. Further studies are required to elucidate the mechanism of PCBs action on ovarian steroidogenesis. PMID- 11016875 TI - National, state, and urban area vaccination coverage levels among children aged 19-35 months--United States, 1998. AB - PROBLEM/CONDITION: High vaccination levels in the population are necessary to decrease disease transmission and prevent disease; therefore, an important component of the U.S. vaccination program is the assessment of vaccination coverage. Current goals are for > or = 90% coverage with recommended vaccines during the first 2 years of life. REPORTING PERIOD: January-December 1998. DESCRIPTION OF SYSTEMS: The National Immunization Survey (NIS) is an ongoing, random-digit-dialed telephone survey that gathers vaccination coverage data for children aged 19-35 months in all 50 states and 28 urban areas. Vaccination coverage rates derived from NIS data are adjusted statistically for households with multiple telephone lines, household nonresponse, the proportion of households without telephones, and vaccination provider nonresponse. The results were also adjusted to match the known total population of children in each survey area. RESULTS: On the basis of NIS data, national coverage was > or = 90% for three doses of poliovirus vaccine (Polio), three doses of Haemophilus influenzae type b vaccine (Hib), and one dose of measles-containing vaccine (MCV). Coverage was the highest ever reported for four doses of any diphtheria and tetanus toxoids and pertussis vaccine (DTP) (i.e., diphtheria and tetanus toxoids and pertussis vaccine, diphtheria and tetanus toxoids [DT], or diphtheria and tetanus toxoids and acellular pertussis vaccine [DTaP]) (83.9%), three doses of hepatitis B vaccine (Hep B, 87.0%), and one dose of varicella vaccine (43.2%). The number of states achieving the > or = 90% goal was 47 for three doses of Hib, 40 for three doses of Polio, 40 for one dose of MCV, nine for three doses of Hep B, and seven for four doses of DTP. Proportionally fewer urban areas achieved the > or = 90% goal: 23 of 28 for three doses of Hib, 13 for three doses of Polio, 16 for one dose of MCV, five for three doses of Hep B, and one for four doses of DTP. No state or urban area has yet achieved the > or = 90% goal for varicella. INTERPRETATION: Findings from the 1998 NIS indicate that national vaccination coverage levels for routinely recommended childhood vaccines are at the highest levels ever reported. However, substantial variation in coverage remains at the state and urban area levels. PUBLIC HEALTH ACTIONS: The public health community and vaccination providers in areas with low coverage should intensify their efforts to implement recommended strategies for increasing vaccination coverage to ensure that children are equally well protected throughout the United States. PMID- 11016876 TI - Vaccination coverage among children enrolled in Head Start programs or day care facilities or entering school. AB - PROBLEM/CONDITION: Undervaccinated children enrolled in day care centers and schools are vulnerable to outbreaks of vaccine-preventable diseases. A Healthy People 2000 objective is to increase to > or = 95% vaccination coverage among children attending licensed day care facilities and kindergarten through postsecondary school (objective 20.11). REPORTING PERIOD COVERED: September 1997 June 1998. DESCRIPTION OF SYSTEM: CDC's National Immunization Program administers grants to support 64 vaccination programs. These programs are in all 50 states, eight territories or jurisdictions (American Samoa, Republic of Marshall Islands, Federated States of Micronesia, Guam, Commonwealth of Northern Mariana Islands, Puerto Rico, Republic of Palau, and the U.S. Virgin Islands), five cities (Chicago, Houston, San Antonio, New York City, and Philadelphia), and the District of Columbia. Grant guidelines require annual school vaccination surveys and biennial surveys of Head Start programs and licensed day care facilities. This system constitutes the only source of nationally representative vaccination coverage estimates for these populations. RESULTS: Head Start Programs: Of the 64 reporting areas, 33 (51.6%) submitted coverage levels for children enrolled in Head Start programs. Of these, all 33 programs reported coverage levels for diphtheria and tetanus toxoids and pertussis vaccine (DTP), diphtheria and tetanus toxoids (DT), or tetanus toxoids (Td), poliovirus vaccine, and measles vaccine; and 32 reported coverage levels for mumps and rubella vaccines. Four programs reported coverage levels for the combined measles, mumps, and rubella vaccine (MMR). The mean vaccination coverage levels for the 1997-98 school year among the reporting vaccination programs were 97.8% for poliovirus vaccine (range: 80.0%-100.0%), 97.0% for DTP/DT/Td (range: 87.7%-100.0%), 93.3% for measles vaccine (range: 91.4%-100.0%), and 93.2% for mumps and rubella vaccines (range: 91.4%-100.0%). Licensed Day Care Facilities: Of the 63 reporting areas with licensed day care facilities, 38 (60.3%) submitted coverage levels for enrolled children. Of these, all 38 programs reported coverage levels for poliovirus vaccine and DTP/DT/Td, 37 reported coverage levels for measles vaccine, and 36 reported coverage levels for mumps and rubella vaccines. Four programs reported coverage levels for the combined MMR. The mean vaccination coverage levels among the reporting areas were 95.8% for poliovirus vaccine (range: 85.1%-99.8%), 95.7% for DTP/DT/Td (range: 77.6%-99.9%), 89.1% for measles vaccine (range: 78.0%-99.9%), and 89.1% for mumps and rubella vaccines (range: 78.0%-99.9%). Kindergarten/First Grade: Of the 64 reporting areas, 43 (67.2%) submitted coverage levels for children enrolled in kindergarten and first grade. Of these 43 programs, 42 reported coverage levels for poliovirus vaccine and DTP/DT/Td, and 43 reported coverage levels for measles, mumps, and rubella vaccines. Four of the 43 programs reported coverage levels for the combined MMR. The mean vaccination coverage levels among the reporting areas were 96.7% for poliovirus vaccine (range: 82.8%-99.9%), 96.7% for DTP/DT/Td (range: 82.8% 99.8%), 96.0% for measles vaccine (range: 82.8%-99.9%), and 96.5% for mumps and rubella vaccines (range: 82.8%-99.9%). INTERPRETATION: High levels of vaccination coverage among children entering school most likely result from the successful implementation of state-specific school vaccination laws, which have applied to children entering school in all states and the District of Columbia since at least 1990. All states, territories, and the District of Columbia have additional laws that require vaccination of children in licensed day care facilities. However, because a high proportion of states and territories did not submit vaccination coverage reports to CDC, these estimated means may not reflect levels for all children in the United States. PMID- 11016877 TI - Influenza, pneumococcal, and tetanus toxioid vaccination of adults--United States, 1993-7. AB - PROBLEM/CONDITION: An increasing proportion of adults have received recommended vaccinations against influenza, pneumococcal infection, and tetanus. However, in 1995, fewer than 60% of adults were vaccinated as recommended. REPORTING PERIOD COVERED: 1993-1997. DESCRIPTION OF SYSTEM: Data were obtained from the state based Behavioral Risk Factor Surveillance System (BRFSS) for 1993, 1995, and 1997 and from the National Health Interview Survey (NHIS) for 1995 to describe national, regional, and state-specific patterns of use of influenza and pneumococcal vaccines and tetanus toxoid among noninstitutionalized adults aged > or = 18 years. RESULTS: Among adults aged > or = 65 years in 1995, 58% reported receiving an influenza vaccination during the previous 12 months, and 34% reported ever receiving a pneumococcal vaccination. In this age group, non Hispanic whites were more likely to report receipt of influenza (61%) and pneumococcal vaccines (36%) than non-Hispanic blacks (40% and 22%, respectively) and Hispanics (50% and 23%, respectively). Among the 50 states and the District of Columbia, the median vaccination level among older adults (i.e., persons aged > or = 65 years) increased from 51% in 1993 to 66% in 1997 for influenza vaccine, and from 28% in 1993 to 46% in 1997 for pneumococcal vaccine. Adults with chronic medical conditions had low vaccination levels. Those aged 50-64 years were more likely than those aged 18-49 years to report influenza (38% versus 20%) and pneumococcal vaccination (20% versus 12%). In 1995, the proportion of adults who reported receiving a tetanus vaccination during the previous 10 years decreased with age, from 65% among those aged 18-49 years to 54% among those aged 50-64 years and to 40% among those aged > or = 65 years. In each age group, women were less likely than men to report receiving tetanus toxoid; and among adults aged > or = 65 years, Hispanics and Asians/Pacific Islanders were least likely among all racial/ethnic groups to report receiving tetanus toxoid. INTERPRETATION: By 1995, the Healthy People 2000 objective to increase to at least 60% the proportion of persons aged > or = 65 years who had received annual influenza vaccination had been achieved among non-Hispanic whites (objective 20.11). However, substantial improvement is needed among non-Hispanic blacks, Hispanics, and adults aged < 65 years with high-risk medical conditions. PUBLIC HEALTH ACTIONS: Continued surveillance of vaccine coverage among adults will direct attention to undervaccinated populations that may be disproportionately affected by vaccine preventable diseases. Vaccination coverage data can be used to guide efforts to increase awareness among health-care providers and the public about the benefits of vaccination, establish systems to ensure that every contact with the health care system is used to update vaccinations, and further support financial mechanisms to increase vaccine delivery. PMID- 11016878 TI - Proteinase inhibitors and activators strategic targets for therapeutic intervention. AB - The contributions in this meeting represent the state of the art in study of proteinases and their biological and therapeutic regulation. The near future of proteinase research will be fashioned by the new horizons of proteomics research [61], adding substance to the genomic data. Even new approaches in drug discovery, such as combinatorial chemistry, impact upon the understanding of the proteinase function, as with the discovery of a novel allosteric exosite in FVIIa by probing with libraries of ligands [62]. What is without question is that proteinases will remain at the forefront of understanding and intervention in human biochemistry and human disease pathology. PMID- 11016879 TI - Receptor-mediated regulation of plasminogen activator function: plasminogen activation by two directly membrane-anchored forms of urokinase. AB - The generation of the broad specificity serine protease plasmin in the pericellular environment is regulated by binding of the urokinase-type plasminogen activator (uPA) to its specific glycosylphosphatidylinositol (GPI) anchored cell-surface receptor, uPAR. This interaction potentiates the reciprocal activation of the cell-associated zymogens pro-uPA and plasminogen. To further study the role of uPAR in this mechanism, we have expressed two directly membrane anchored chimeric forms of uPA, one anchored by a C-terminal GPI-moiety (GPI uPA), the other with a C-terminal transmembrane peptide (TM-uPA). These were expressed in the monocyte-like cell lines U937 and THP-1, which are excellent models for kinetic and mechanistic studies of cell-surface plasminogen activation. In both cell-lines, GPI-uPA activated cell-associated plasminogen with characteristics both qualitatively and quantitatively indistinguishable from those of uPAR-bound uPA. By contrast, TM-uPA activated cell-associated plasminogen less efficiently. This was due to effects on the K, for plasminogen activation (which was increased up to five-fold) and the efficiency of pro-uPA activation (which was decreased approximately four-fold). These observations suggest that uPAR serves two essential roles in mediating efficient cell-surface plasminogen activation. In addition to confining uPA to the cell-surface, the GPI anchor plays an important role by increasing accessibility to substrate plasminogen and, thus, enhancing catalysis. However, the data also demonstrate that, in the presence of an alternative mechanism for uPA localization, uPAR is dispensable and, therefore, unlikely to participate in any additional interactions that may be necessary for the efficiency of this proteolytic system. In these experiments zymogen pro-uPA was unexpectedly found to be constitutively activated when expressed in THP-1 cells, suggesting the presence of an alternative plasmin-independent proteolytic activation mechanism in these cells. PMID- 11016880 TI - Bradykinin analogues with beta-amino acid substitutions reveal subtle differences in substrate specificity between the endopeptidases EC 3.4.24.15 and EC 3.4.24.16. AB - The closely related zinc metalloendopeptidases EC 3.4.24.15 (EP24.15) and EC 3.4.24.16 (EP24.16) cleave many common substrates, including bradykinin (BK). As such, there are few substrate-based inhibitors which are sufficiently selective to distinguish their activities. We have used BK analogues with either alanine or beta-amino acid (containing an additional carbon within the peptide backbone) substitutions to elucidate subtle differences in substrate specificity between the enzymes. The cleavage of the analogues by recombinant EP24.15 and EP24.16 was assessed, as well as their ability to inhibit the two enzymes. Alanine substituted analogues were generally better substrates than BK itself, although differences between the peptidases were observed. Similarly, substitution of the four N-terminal residues with beta-glycine enhanced cleavage in some cases, but not others. beta-Glycine substitution at or near the scissile bond (Phe5-Ser6) completely prevented cleavage by either enzyme: interestingly, these analogues still acted as inhibitors, although with very different affinities for the two enzymes. Also of interest, beta-Gly8-BK was neither a substrate nor an inhibitor of EP24.15, yet could still interact with EP24.16. Finally, while both enzymes could be similarly inhibited by the D-stereoisomer of beta-C3-Phe5-BK (IC50 approximately 20 microM, compared to 8 microM for BK), EP24.16 was relatively insensitive to the L-isomer (IC50 12 approximately microM for EP24.15, >40 microM for EP24.16). These studies indicate subtle differences in substrate specificity between EP24.15 and EP24.16, and suggest that beta-amino acid analogues may be useful as templates for the design of selective inhibitors. PMID- 11016881 TI - Identification of chymotrypsin inhibitors from a second-generation template assisted combinatorial peptide library. AB - In an earlier study (McBride JD, Freeman N, Domingo GJ, Leatherbarrow RJ. Selection of chymotrypsin inhibitors from a conformationally-constrained combinatorial peptide library. J. Mol. Biol. 1996; 259: 819-827) we described a resin-bound cyclic peptide library, constructed based on the sequence of the anti tryptic reactive site loop of Bowman Birk Inhibitor (BBI), a proteinase inhibitor protein. This library was used to identify re-directed chymotrypsin inhibitors with Ki values as low as 17 nM. We have now extended this work by constructing an enhanced library in which a further position, at the P4 site of the inhibitor, has been randomized. This new library has variation at three target locations (P4, P1 and P2) within the inhibitory loop region, producing 8,000 variants. Screening this library allowed selection of new inhibitor sequences with Ki values as low as 3.4 nM. The success of this approach is reflected by the fact that the inhibition constant given by the selected peptide sequence is slightly lower than that reported against chymotrypsin for the most studied full length BBI protein, Soybean BBI 2-IV. PMID- 11016882 TI - Assessing the protease and protease inhibitor content of the human genome. AB - The revealing of the entire complement of protease and protease inhibitor sequences by the Human Genome Project will be of great importance to both academic and pharmaceutical research. Although the finishing phase is not yet complete, a selection of secondary annotation sources and comparisons with completed model organism genomes already allow useful estimates to be made. Conservative extrapolation suggests a total of approximately 1.8% for human proteases. This is close to the figures for yeast (1.7%) and worm (1.8%) but lower than the fly (3.4%) which has a large trypsin-like protease content. Using estimates for the human proteome of between 40,000 and 60,000 genes would extrapolate to 700-1,100 proteases, compared with approximately 360 currently represented as GenBank mRNAs. Preliminary comparisons between domain annotations for predicted human gene products and completed proteins suggest the genomic protease family and mechanistic class distributions will broadly reflect those in the current transcript data. The protease:inhibitor ratio at the mRNA level is currently approximately 9:1, but genome annotation data indicate that inhibitory domains are more widespread than this ratio would indicate. PMID- 11016883 TI - The use of seldi proteinchip arrays to monitor production of Alzheimer's betaamyloid in transfected cells. AB - beta-Amyloid (Abeta), a 39-43 residue peptide, is the principal component of senile plaques found in the brains of patients with Alzheimer's disease (AD). There are two main lines of evidence that its deposition is the cause of neurodegeneration. First, mutations found in three genes in familial Alzheimer's cases give rise to increased production of the longest, most toxic, form, Abeta 1 42. Second. aggregated Abeta is toxic to neuronal cells in culture. Inhibitors of the proteases involved in its release from the amyloid precursor protein are, therefore, of major therapeutic interest. The best candidates for the releasing proteases are both aspartyl proteases, which are integrated into the membranes of the endoplasmic reticulum and Golgi network. A sensitive assay using Ciphergen's Seldi system has been developed to measure all the variants of Abeta in culture supernatants, which will be of great value in screening inhibitors of these proteases. With this assay, it has been shown that increasing intracellular cholesterol increases the activities of both beta-secretase, and gamma-secretase 42. Moreover, changing the intracellular targeting of amyloid precursor glycoprotein (APP) yields increased alpha-secretase cleavage, and increases in the amounts of oxidized/nitrated forms of Abeta. PMID- 11016884 TI - Design and synthesis of inhibitors incorporating beta -amino acids of metalloendopeptidase EC 3.4.24.15. AB - Endopeptidase EC 3.4.24.15 (EP 24.15) is a thermolysin-like metalloendopeptidase which is expressed widely throughout the body, with the highest concentrations in the brain, pituitary and testis. While the precise role of EP 24.15 remains unknown, it is thought to participate in the regulated metabolism of a number of specific neuropeptides. Of the limited number of inhibitors described for EP 24.15, N-[1-(R,S)-carboxy-3-phenylpropyl]-Ala-Ala-Tyr-p-amino benzoate (CFP) is the most widely studied. CFP is a potent and specific inhibitor, but is unstable in vivo due to its cleavage between the alanine and tyrosine residues by the enzyme neprilysin (EP 24.11). The cpp-Ala-Ala N-terminal product of this cleavage is a potent inhibitor of angiotensin converting enzyme, which further limits the use of CFP in vivo. To generate specific inhibitors of EP 24.15 that are resistant to in vivo proteolysis by EP 24.11, beta-amino acids have been incorporated into the structure of CFP. We have prepared racemic mixtures of beta amino acids containing proteogenic side chains, which are 9 fluorenylmethoxycarbonyl (Fmoc)-protected, and several analogues of CFP containing beta-amino acids have been synthesized by solid phase peptide synthesis. The results of stability and inhibitory studies of these new analogues show that the incorporation of beta-amino acids adjacent to the scissile bond can indeed stabilize the peptides against cleavage by EP 24.11 and still inhibit EP 24.15. The results obtained in these studies demonstrate the potential of these amino acids in the synthesis of peptidomimetics and in the design of new stable and specific therapeutics. PMID- 11016885 TI - Proteasome inhibitors: from in vitro uses to clinical trials. AB - Proteasomes are multicatalytic proteinase complexes which play a central role in intracellular protein degradation. They catalyse key events in cell cycle regulation and in the activation of the transcription factor NFkappaB. Proteasome inhibitors have been useful for the characterization of proteasome catalytic components and in the elucidation of proteasome functions in animal cells. Potent small peptide inhibitors of proteasomes also represent a novel approach to the treatment of inflammatory diseases (which involve activation of NFkappaB) and cancer. Such compounds have recently been shown to be effective in a variety of animal models, and at least one is currently in use in clinical trials. PMID- 11016886 TI - Design and synthesis of acidic dipeptide hydroxamate inhibitors of procollagen C proteinase. AB - Procollagen C-proteinase (PCP) is essential for the cleavage of procollagen to collagen in the extracellular matrix of animals and is, therefore, of major relevance to studies of ectopic deposition of collagen during fibrosis. In this study, we describe the design and synthesis of acidic side chain hydroxamate dipeptide inhibitors of PCP having IC50 values in the range 0.1-10 microM that mimic the location of aspartic acid residues in the P1' and P2' positions (i.e. immediately C-terminal) of the PCP cleavage site in procollagen. Assays of PCP using purified human type I procollagen (a natural substrate of PCP) showed that the structure activity relationship of the inhibitors was improved with a glutamic acid mimic at the P1' position. The results also showed that the presence of an acidic side chain at the P2' position was not necessary for PCP inhibition. Marimastat and BB3103, which are highly effective inhibitors of matrix metalloproteinases and ADAMS proteinases, respectively, did not inhibit PCP. PMID- 11016887 TI - Variation of lipids and lipoproteins in premenopausal women compared with men and postmenopausal women. DELTA (Dietary Effects on Lipoproteins and Thrombogenic Activity) Investigators. AB - Numerous studies have reported cyclic fluctuations in lipids and lipoproteins as a function of the phase of the menstrual cycle. However, the reported patterns are quite variable and have led to an unclear picture of the influence of the menstrual cycle on the variability of lipids, and hence of the role of the menstrual cycle phase in the interpretation of serum lipids for premenopausal women. As part of the DELTA Study (Dietary Effects on Lipoproteins and Thrombogenic Activity), we evaluated the cyclic variation of circulating lipids and lipoproteins in 39 premenopausal women and compared intraindividual variances in these women, 18 postmenopausal women, and 46 men under conditions of tight dietary control. Cholesterol, high-density lipoprotein (HDL)-cholesterol, low density lipoprotein (LDL)-cholesterol, triglyceride, apolipoproteins A-1 (apo A 1) and B-100 (apo B-100), and lipoprotein (a) [Lp(a)] all demonstrated cycling in the premenopausal women. However, the observed cycling accounts for only a small fraction of the total biologic variability of lipids in premenopausal women. The magnitude of total intraindividual variability based on coefficient of variation (CV) for these lipids in premenopausal women (CV, 4% to 8.1%) was similar to that found for men (CV, 4.3% to 9.1%) and for postmenopausal women (CV, 3.7% to 6.7%). These results suggest that protocols for screening and monitoring of serum lipids in premenopausal women need not differ from those used for men or postmenopausal women. PMID- 11016888 TI - Masoprocol decreases serum triglyceride concentrations in rats with fructose induced hypertriglyceridemia. AB - Historically, extracts of the creosote bush have been used by native healers of the Southwest region of North America to treat symptoms of type 2 diabetes. More recently, we have shown that masoprocol (nordihydroguaiaretic acid), a pure compound isolated from the creosote bush (Larrea tridentata), decreases serum glucose and triglyceride (TG) levels when administered orally in rodent models of type 2 diabetes. The present studies were undertaken to determine if masoprocol also decreases TG concentrations in rats with fructose-induced hypertriglyceridemia (HTG), a nondiabetic model of HTG associated with insulin resistance and hyperinsulinemia. Serum TG levels, which were significantly higher after rats ate a fructose-enriched (60% by weight) diet for 14 days as compared with chow-fed controls (411 v 155 mg/dL, P < .01), decreased in a stepwise fashion in fructose-fed rats treated orally with masoprocol for 4 to 8 days over a dose range of 10 to 80 mg/kg twice daily. Using the nonionic detergent Triton WR 1339 to compare TG secretion rates in masoprocol- and vehicle-treated rats, masoprocol at a dose of 40 or 80 mg/kg twice daily, significantly reduced hepatic TG secretion (P < .01) and liver TG content (P < .001), whereas lower doses of masoprocol decreased serum TG without an apparent reduction in hepatic TG secretion. Administration of Intralipid (a fat emulsion) showed that the half time for removal of TG from serum was also shorter in masoprocol-treated rats versus vehicle-treated controls (31 v 64 minutes, P < .05). In addition adipose tissue lipoprotein lipase (LPL) activity was increased in masoprocol-treated rats and adipose tissue hormone-sensitive lipase (HSL) activity was decreased. We conclude that masoprocol administration to rats with fructose-induced HTG results in lower serum TG levels associated with reduced hepatic TG secretion and increased peripheral TG clearance. PMID- 11016889 TI - Dietary effects on insulin and nutrient metabolism in mesenteric lymph node cells, splenocytes, and pancreatic islets of BB rats. AB - The present studies were performed to determine if a protective diet has different effects on the metabolic activity or function of islet cells, as well as the metabolic activity of mesenteric lymph node (MLN) cells and spleen cells, from BioBreeding (BB) rats. Diabetes-prone BB (BBdp) rats and control non diabetes-prone BB (BBc) rats were fed for about 20 days either a mainly plant based diabetogenic diet, NIH-07 (NIH), or a protective semipurified diet with hydrolyzed casein (HC) as the amino acid source. At 6 to 8 weeks of age, BBdp rats had high plasma D-glucose and low insulin concentrations, low insulin content, and low metabolic and secretory responses to D-glucose in isolated pancreatic islets. Islet metabolism, as measured by accumulation of 14C-acidic metabolites, amino acids, and the ratio of D-[U-14C]glucose oxidation and D-[5 3H]glucose utilization was increased in control rats fed HC (P < .05); a similar trend in BBdp rats was not significant. Feeding the HC diet increased islet insulin content (P < .01) by 13% in BBdp and 23% in BBc rats; other metabolic and hormonal variables were unaffected. Compared with BBc rats, BBdp rats displayed higher rates of L-[U-14C]glutamine oxidation, D-[5-3H]glucose utilization, and D [U-14C]glucose oxidation in MLN cells, but not in splenocytes. There was a dramatic decrease of L-[U-14C]glutamine oxidation in MLN cells from BBc and BBdp rats fed HC. Glycolysis was decreased in control rats. We conclude that the protection afforded by feeding BBdp rats a HC diet is associated with increased insulin in target beta cells and downregulation of metabolic activity in gut associated MLN cells. Metabolic activity in splenocytes, cells representative of the systemic immune system, was less affected. These data suggest that diet induced metabolic changes occur in the islets and nearby cells of the gut immune system in the period before classic insulitis. Changes in the islets were smaller in comparison to the dramatic remodeling of nutrient catabolism in MLN cells. MLN downregulation may reflect baseline metabolic activity in the absence of diabetogenic (or other) food antigens and further highlights an important interaction between diabetogenic food antigens and the gut immune tissues. PMID- 11016890 TI - Decreased insulin production and increased insulin sensitivity in the klotho mutant mouse, a novel animal model for human aging. AB - We have recently identified a novel gene, klotho (kl), which may suppress several aging phenotypes. A defect of kl gene expression in the mouse results in a syndrome resembling human aging, such as arteriosclerosis, skin atrophy, osteoporosis, and pulmonary emphysema. To determine whether mouse homozygotes for the kl mutation (kl/kl) show abnormal glucose metabolism, an oral glucose tolerance test (OGTT) was performed at 6 to 8 weeks of age. Blood glucose levels during the OGTT were significantly lower in kl/kl mice versus wild-type mice. The insulin content of the pancreas was significantly lower in kl/kl mice compared with wild-type mice. Decreased insulin production was also supported by Northern blot analysis showing lower levels of insulin mRNA in kl/kl mice. To examine how lower blood glucose levels may exist in kl/kl mice despite decreased insulin production, insulin tolerance tests (ITTs) were performed. The glucose decline following insulin injection was more severe in kl/kl mice versus wild-type mice, suggesting that insulin sensitivity was higher in kl/kl mice versus wild-type mice. In kl/kl mice, an augmented expression of GLUT4 in skeletal muscle was demonstrated by both Northern blot analysis and Western blot analysis. Thus, we conclude that insulin production is decreased and insulin sensitivity is increased in the klotho mouse, a novel animal model for human aging. PMID- 11016891 TI - Estradiol and testosterone effects on lipids in black and white boys aged 10 to 15 years. AB - Previous studies of lipids in adolescent males have shown greater increases in triglycerides and decreases in high-density lipoprotein cholesterol (HDL-C) in white boys compared with black boys, significant correlations between sex hormones and lipids, and complex body mass index (BMI) hormone-lipid associations. Within this frame of reference, we assessed race, BMI, and sex hormones as predictors of lipid parameters in 536 black and white boys recruited from area schools. Black boys were more advanced in puberty than white boys. After adjusting for pubertal stage, estradiol (E2) levels were higher in black boys but free testosterone (T) levels did not differ. Age, pubertal stage, race, BMI, free T, and E2 were entered as explanatory variables for lipids in backward stepwise regression analyses. The BMI and race were retained in every model. Black boys had lower triglycerides and apolipoprotein B (apo B) and higher HDL-C. E2 was inversely associated with total cholesterol (TC), triglycerides, low density lipoprotein cholesterol (LDL-C), apo B, and the LDL-C/HDL-C ratio. Free T was inversely associated with HDL-C and positively associated with apo B. Given the increases in free T and E2 during adolescence and the association of these hormones with both atherogenic and protective lipid parameters, racial differences in E2 could contribute to the more atherogenic lipid profile found in white boys after puberty. PMID- 11016892 TI - Evidence for dramatically increased bone turnover in spontaneously hypertensive rats. AB - Using the 3H-tetracycline model, whole-body skeletal bone resorption was compared among male and female spontaneously hypertensive (SHR) rats and normotensive Wistar Kyoto (WKy) and Sprague-Dawley (SD) rats. Immature animals undergoing rapid skeletal growth and bone sculpting showed a tendency for decreased indices of skeletal resorption in females compared with males. By 24 weeks of age, the indices of the rate of resorption and extent of metabolically reactive bone in male rats were decreased a mean of 68% and 74%, respectively, compared with values obtained at 8 weeks. By comparison, values for 24-week-old females decreased only 26% and 56%, respectively, evidence for a significantly elevated level of resorptive activity in mature females compared with males in each of the 3 rat strains. Within-sex comparisons of 24-week-old animals indicated that bone resorptive activity was similar between normotensive male and normotensive female groups. By comparison, the resorptive activity was significantly increased in both male and female hypertensive rats compared with normotensive controls. This condition was exaggerated in female hypertensive rats, which showed an approximate 81% and 44% increase in the indices of rate of resorption and extent of metabolically reactive bone compared with normotensive WKy controls. The results indicate a marked sexual dichotomy in the decline of skeletal bone resorptive activity following maturation and slowing of skeletal growth. They further indicate a significant elevation of whole skeleton bone turnover in male SHR rats and dramatically increased bone turnover in female SHR rats. PMID- 11016893 TI - Changes in calciotropic hormones and biochemical markers of bone metabolism in patients with human immunodeficiency virus infection. AB - Data on the bone metabolism of human immunodeficiency virus (HIV)-infected patients are still extremely rare. To investigate the influence of HIV infection on the calciotropic hormones and markers of bone metabolism, we therefore performed a cross-sectional study on 100 patients (65 males and 35 females) with proven HIV infection. The following criteria were used for exclusion from the study: age less than 20/more than 50 years, confinement to bed, wasting symptoms, treatment with agents containing ketoconazole, renal or hepatic insufficiency, clinical or echographic signs of liver cirrhosis, endocrine diseases, or treatment with medications known to influence bone metabolism. Bone mineral content (BMC) was determined by single-photon absorptiometry on the left forearm. Reduced BMC was found among the male and female HIV-infected patients. Additional long-term use of heroin resulted in a severe loss of mineralization in the respective females. The markers of bone metabolism were determined in urine and serum samples. Significantly lower osteocalcin concentrations were found, indicating a reduced bone formation rate whose severity showed a significant correlation with the progressive loss of CD4 helper cells and was independent of low vitamin D3 levels (1,25-dihydroxycholecalciferol) and alterations of protein metabolism. Increased urinary excretion of cross-links as an expression of enhanced bone resorption was likewise significantly correlated with the loss of CD4 helper cells and independent of the vitamin D concentration and protein metabolism. It is therefore concluded that the changes in bone metabolism are mainly due to mechanisms of the impaired immune defense of HIV-infected patients. PMID- 11016894 TI - Features of the metabolic syndrome in the spontaneously hypertriglyceridemic Wistar Ottawa Karlsburg W (RT1u Haplotype) rat. AB - The aim of this study was to characterize the Wistar Ottawa Karlsburg W ([WOKW] RT1u haplotype) rat in a cross-sectional study (up to 14 weeks of age) for traits with pathophysiological relevance to the metabolic syndrome in comparison to the Dark Agouti (DA) rat, to determine the age at which the WOKW rat begins to manifest the characteristics of the metabolic syndrome. The findings indicate that the WOKW rat is dyslipidemic (high serum triglycerides and low high-density lipoprotein [HDL] cholesterol), hyperinsulinemic, and obese. The interval between 8 and 10 weeks appears to be the crucial age after which the most dramatic changes were observed in the measured phenotypic traits in the WOKW rat, as well as the most expressive differences between the WOKW and DA strains. Considering the phenotypic differences between WOKW and DA rats, the DA rat provides an appropriate control strain for crossing studies with the WOKW rat, which might contribute to the explanation of the genetic basis for traits of the metabolic syndrome in this model. PMID- 11016895 TI - Tyrosine hydroxylase gene microsatellite polymorphism associated with insulin resistance in depressive disorder. AB - A high association between type 2 diabetes mellitus and depressive illness has been reported. Insulin resistance during depressive illness might contribute to the linkage between depression and type 2 diabetes. To determine whether the genetic polymorphisms of the tyrosine hydroxylase ([TH] HUMTH01) and insulin (INS VNTR) genes contribute to insulin resistance in depressive illness, we analyzed the association between the polymorphisms and insulin resistance in 41 Japanese patients with depressive disorder, 204 normal control subjects, 161 cohort subjects with normal glucose tolerance (NGT) and without depressive symptomatology, and 59 NGT subjects with depressive symptomatology. The depressive patients had a significantly lower insulin sensitivity index (SI) than the control subjects (P= .016). Depressive NGT subjects had a significantly higher homeostasis model assessment (HOMA) insulin resistance index [HOMA(R)] than the nondepressive NGT subjects (P < .0001). The depressive patients and NGT subjects had more HUMTH01 allele 7 (TH7) than the controls and nondepressive NGT subjects. SI was significantly lower in patients with the TH7/7 homozygote versus patients with the other genotypes and the controls. TH7 was associated with higher HOMA(R) as compared with the other alleles in the NGT subjects. Insulin resistance was associated with depressive disorders. The HUMTH01 and INS-VNTR were associated with insulin resistance and depressive symptoms. PMID- 11016896 TI - Postprandial lipemia after short-term variation in dietary fat and carbohydrate. AB - Replacement of dietary fat with carbohydrate may not reduce the overall risk of coronary heart disease (CHD), because this elevates plasma triacylglycerol (TAG) concentrations. The lipoproteinemic effects of a high-carbohydrate diet are likely to be more marked shortly after the initiation of such a diet than after longer periods of intervention during which adaptive processes may counteract the initial effects. Therefore, we studied the postprandial responses to a standard meal after 3-day dietary intervention periods. An additional objective was to establish a model for future study of the mechanisms involved. Nine normolipidemic men consumed the meal (1.2 g fat, 1.1 g carbohydrate, and 0.2 g protein per 1 kg body mass) after 3 days on a high-carbohydrate diet (68% +/- 3% energy from carbohydrate, mean +/- SD) and also after 3 days on an isoenergetic high-fat diet (66% +/- 5% energy). Venous blood samples were obtained from fasted subjects and for 6 hours after the meal. In the fasted state, TAG was higher after the high-carbohydrate diet (1.18 +/- 0.18 v0.62 +/- 0.09 mmol/L, mean +/- SEM, P = .02) and high-density lipoprotein (HDL) cholesterol was lower (1.01 +/- 0.08 v 1.10 +/- 0.09 mmol/L, P = .002). The area under the plasma TAG concentration versus time curve was 42% +/- 7% higher after the high-carbohydrate diet (P = .003). After the high-carbohydrate diet, the postprandial insulin response did not differ between trials, but glucose and 3-hydroxybutyrate responses were lower (P = .009 and P = .02, respectively) and the lactate response was higher (P = .001). Plasma nonesterified fatty acids (NEFAs) were lower after the high-carbohydrate diet in the fasted state and for 4 hours postprandially, but were higher thereafter (interaction of time x trial, P = .001). These results indicate that compared with a high-fat diet, the plasma TAG response to a standard high-fat meal is markedly higher after a few days on a high-carbohydrate diet, with major differences in the associated metabolic milieu. The magnitude of these changes and the rapidity with which they developed suggest that this model may be attractive for future studies of the underlying mechanisms. PMID- 11016897 TI - Insulin secretion, inositol phosphate levels, and phospholipase C isozymes in rodent pancreatic islets. AB - During a dynamic perifusion, 20 mmol/L glucose, 20 mmol/L alpha-ketoisocaproate (KIC) or 20 mmol/L methyl pyruvate (MP) stimulate biphasic insulin secretory responses from collagenase-isolated rat islets. Peak first-phase insulin responses were comparable for all 3 nutrient agonists. The largest second-phase insulin secretory response was evoked by 20 mmol/L glucose (30-fold above basal release rates), and this response was more sustained than that observed with either 20 mmol/L KIC or 20 mmol/L MP. When mouse islets were perifused under similar conditions, KIC stimulated the largest first-phase insulin response, while comparable acute insulin secretion rates were obtained with glucose- or MP stimulated islets. In contrast to rat islets, the sustained second phase of insulin secretion from mouse islets was minimal regardless of the nutrient secretagogue used. This anomalous response of mouse islets as compared with rat islets could not be ascribed to any obvious difference in the glucose usage rate or insulin content between these 2 species. Glucose, KIC, or MP stimulated significant increases in 3H-inositol phosphates in rat islets. Significantly smaller increases were measured in mouse islets. Comparative Western blot analyses showed pronounced species differences in the expression of phospholipase Cbeta1 (PLCbeta1), PLCbeta2, PLCbeta3, and PLCdelta1 but not PLCgamma1 or protein kinase Calpha (PKCalpha) between rat and mouse islets. PLCbeta4 or PLCdelta2 could not be identified in either species. These findings are consistent with the concept that the underexpression of the nutrient-activated PLC isozyme may account for the minimal inositol phosphate (IP) and second-phase insulin secretory response from mouse islets. PMID- 11016898 TI - Is the erythrocyte membrane fatty acid composition a valid index of skeletal muscle membrane fatty acid composition? AB - Recent studies suggest that insulin sensitivity is related to the fatty acid composition of phospholipids in skeletal muscle (SM) membranes. Since it is difficult to obtain muscle biopsies, it may be useful to have information on the fatty acid composition using more accessible cells such as erythrocytes. This would be possible only if the composition of erythrocyte and muscle membranes are very similar. Since no comparative data are available, we evaluated the phospholipid fatty acid composition of erythrocyte and SM membranes in 16 individuals, 10 nondiabetics (male to female ratio, 4:6; age, 50 +/- 11 years; body mass index, 27 +/- 5 kg/m2; mean +/- SD) and 6 type 2 diabetic patients (male to female ratio, 2:4; age, 64 +/- 5 years; body mass index, 27 +/- 4 kg/m2). All patients underwent abdominal surgery, during which a biopsy of the abdominal rectus muscle (50 to 100 mg) was obtained. Erythrocyte and SM phospholipid fatty acids were extracted and then methylated; the methyl fatty acids were separated and quantified by gas chromatography. Compared with erythrocyte membranes, muscle membranes showed a significantly higher proportion of omega-6 polyunsaturated fatty acid ([PUFA] 43.0% +/- 3.1% v29.7% +/- 1.6%, P < .001) and lower saturated fatty acid ([SFA] 41.1% +/- 1.5% v 43.4% +/- 1.2%, P < .001), monounsaturated fatty acid ([MUFA] 11.5% +/- 1.7% v 20.0% +/- 1.9%, P < .001), and omega-3 PUFA (3.8% +/- 0.6% v 7.4% +/- 1.0%, P < .001). The greatest increase involved linoleic acid (26.9% +/- 2.8% v 10.3% +/- 1.6%, P < .001), whereas lignoceric acid (0.8% +/- 0.2% v 5.0% +/- 0.6%, P < .001) and oleic acid (10.4% +/- 1.6% v 13.5% +/- 1.3%, P < .001) were significantly lower. These results show that erythrocyte and muscle membrane phospholipid fatty acids are significantly different. Therefore, data on SM membranes cannot be extrapolated on the basis of measures of erythrocyte phospholipid fatty acid composition. PMID- 11016899 TI - Troglitazone prevents and reverses dyslipidemia, insulin secretory defects, and histologic abnormalities in a rat model of naturally occurring obese diabetes. AB - Troglitazone has been shown to improve insulin sensitivity and thereby exert hypoglycemic effects in various animal models and humans with insulin resistance and diabetes. The recently established animal model of naturally occurring obese diabetes, the Otsuka Long-Evans Tokushima fatty (OLETF) rat, has many similarities with human type 2 diabetes mellitus and is characterized by a high degree of insulin resistance. In the present study, we examined the effect of pharmacologic intervention with troglitazone on metabolic and histopathologic changes in OLETF rats. Two groups of rats received a troglitazone-rich diet (200 mg/100 g normal chow) from age 12 weeks (ie, before the onset of diabetes) or 28 weeks (ie, after the onset of diabetes) to age 70 weeks, while a third group received standard rat chow. The addition of troglitazone to the diet did not alter food intake or body weight gain. Troglitazone had no influence on visceral adipose depots, but it significantly reduced fasting glucose, insulin, cholesterol, triglyceride (TG), and free fatty acid (FFA) levels. Troglitazone reduced the insulin resistance and maintained the postglycemic insulin response at a normal level, and thus inhibited the development of insulin insensitivity and frank diabetes in OLETF rats up to 70 weeks of age. The pancreatic wet weight and insulin content were significantly higher in the treated rat groups versus the control rats. The morphologic changes observed in the control rats, such as fibrosis and structural disarrangement of islets, were minimal in the troglitazone-treated rats. Our study demonstrates that troglitazone, albeit at a dosage 10 to 15 times higher than that in humans, not only prevents but also reverses the metabolic derangement and histopathologic changes in genetically determined obese diabetes. PMID- 11016900 TI - Plasma cholesteryl ester transfer protein activity is high in infants and is not affected by thyroid hormones. AB - We investigated the effects of thyroid dysfunction on cholesteryl ester transfer protein (CETP) by studying plasma CETP activity in hypothyroid infants before and after they were rendered euthyroid by L-thyroxine (LT4) replacement therapy. To exclude environmental factors possibly affecting plasma CETP activity, we selected hypothyroid infants to study plasma CETP activity. Plasma CETP activity was measured as the rate of radiolabeled cholesteryl ester transfer from high density lipoprotein (HDL) to serum apolipoprotein B (apo B)-containing lipoproteins in plasma from 14 hypothyroid infants before and 2 months after LT4 replacement, 23 normal infants, and 61 normal adults. Relationships between CETP and thyroid hormones were examined separately in the 14 hypothyroid infants and 32 euthyroid infants, including the 14 above-described hypothyroid infants and an additional 18 treated hypothyroid infants. Serum levels of thyrotropin (TSH), free thyroxine (FT4), and free triiodothyronine (FT3) were also determined on an autoanalyzer system in our hospital. In contrast to previous reports, we found no differences in plasma CETP activity between hypothyroid infants and age-matched normal infants. LT4 substitution did not cause any changes in plasma CETP activity after therapy. Plasma CETP activity showed no correlation with serum TSH, FT4, and FT3 levels. Both hypothyroid and normal infants were found to have significantly higher plasma CETP activity than normal adults. From these results, we conclude that in infants thyroid hormones do not affect plasma CETP activity, and normal infants have high plasma CETP activity, compared with normal adults. PMID- 11016901 TI - Changes in dietary fatty acids modify the decreased lipolytic beta3-adrenergic response to hyperinsulinemia in adipocytes from pregnant and nonpregnant rats. AB - The effect of dietary olive oil and fish oil on the lipolytic dose-response of the beta3-adrenergic agonists, epinephrine, isoproterenol, BRL-37344, and CGP 12177, in adipocytes was studied in pregnant and virgin rats either untreated or under hyperinsulinemic-euglycemic conditions. Rats were fed a semisynthetic diet containing 5% of either olive oil or fish oil and studied at day 20 of treatment and/or gestation. Plasma glucose was lower and plasma insulin, triglycerides, and free fatty acids (FFAs) were higher in pregnant versus virgin rats, and the insulin sensitivity index was lower in the former. Lumbar adipose tissue phospholipid fatty acids showed a significantly higher monounsaturated fatty acid and a lower (n - 3) fatty acid content in rats fed the olive oil diet versus the fish oil diet. The lipolytic dose-response curve of either adrenergic agent was always lower in adipocytes from untreated pregnant rats versus virgin rats, and whereas the hyperinsulinemic-euglycemic clamp decreased these responses in adipocytes from virgin rats fed the olive oil diet only, adipocytes from pregnant rats always showed a decreased dose-response lipolytic curve. Thus, the lipolytic responsiveness of beta3-adrenoceptor (beta3-AR) agonists by adipocytes is impaired in cells from rats made hyperinsulinemic chronically by pregnancy or acutely by the hyperinsulinemic-euglycemic clamp, but such response to the acute condition disappears when the adipocyte phospholipid composition is modified by changes in dietary fatty acids. PMID- 11016902 TI - Soy protein reduces the arterial low-density lipoprotein (LDL) concentration and delivery of LDL cholesterol to the arteries of diabetic and nondiabetic male cynomolgus monkeys. AB - We have previously shown that soy protein consumption improves lipoprotein concentrations and reduces the progression of atherosclerosis in cynomolgus monkeys. The mechanism for these beneficial effects is unclear. The purpose of this study was to determine potential mechanisms for the atheroprotective effects of soy and to determine if these effects extend to diabetic monkeys. We designed an experiment with a 2 x 2 factorial design in which adult male monkeys (N = 23) were fed an atherogenic diet with a protein source of either soy isolate or casein and lactalbumin, and the monkeys were either control or streptozotocin induced diabetic. Diabetics had significantly increased fasting glucose and glycated hemoglobin (GHb) levels; this relationship was not affected by the type of dietary protein. Diabetics also had increased total (TC) and low-density lipoprotein cholesterol (LDLC) concentrations. However, soy consumption significantly reduced TC and LDLC concentrations in both control and diabetic monkeys. Plasma and arterial LDL metabolism was determined by injecting 125I-LDL at 48 hours and 131I-tyramine cellobiose LDL at 1 hour prior to necropsy. This allowed a determination of the arterial LDL concentration, permeability, and arterial LDL delivery. An increase in the whole-body plasma LDL fractional catabolic rate (FCR) was found with soy. Soy significantly reduced the arterial LDL concentration across all arterial sites by an average of 50%. Soy also significantly reduced the delivery of LDLC to all arterial sites by an average of 40%. While this was primarily due to the lower plasma LDLC concentration, LDL permeability in the carotid bifurcation and internal carotid arteries was also reduced. There was no additional effect of diabetes. These beneficial effects on plasma and arterial LDL metabolism would be expected to reduce atherosclerosis and were found in both control and diabetic monkeys. PMID- 11016903 TI - A polymorphism of the 5'-flanking region of the glucocorticoid receptor gene locus is associated with basal cortisol secretion in men. AB - There is growing evidence that cortisol secretion and/or metabolism is associated with cardiovascular risk factors. Previous studies have shown that cardiovascular risk factors are associated with stimulated cortisol secretion and not with basal cortisol secretion. With the restriction enzyme Tth111I, a variant of the 5' flanking region of the glucocorticoid receptor gene locus (GRL) was identified. The genotypes were compared for measurements of cortisol secretion and cardiovascular risk factors in a cohort (N = 284) of randomly selected middle aged men. The frequency of the 3.4/3.4-, 3.4/3.8-, and 3.8/3.8-kilobase (kb) genotypes was 49.6%, 41.4%, and 9.0% respectively. The 3.8-kb homozygotes showed higher total and evening cortisol levels with a trend for elevated levels over the day. Neither stimulated or suppressed cortisol secretion nor anthropometric, endocrine, metabolic, and hemodynamic cardiovascular risk factors were significantly different among the genotypes. Since the polymorphism studied herein is localized at the 5'-flanking region of the GRL, the results suggest that elevated basal cortisol secretion is associated with a polymorphism of the promoter region. PMID- 11016904 TI - Ontogeny of the catalytic subunit and putative glucose-6-phosphate transporter proteins of the rat microsomal liver glucose-6-phosphatase system. AB - The catalytic subunit (p36) and putative glucose-6-phosphate (G6P) transporter (p46) protein levels of the rat glucose-6-phosphatase (G6Pase) system were studied in relation to G6Pase hydrolytic activity and G6P uptake in liver microsomes during the fetal to neonatal period. The mean G6P hydrolytic activity in liver microsomes increased significantly from the 20th to 21st day of gestation (from 6 to 22 mU/mg protein) and was further enhanced by 3-fold 6 hours after birth, with a maximal activity at 1 day of age (112 mU/mg protein). In contrast, G6P uptake into the vesicles was undetectable before birth, appeared after day 1 (656 pmol/mg protein), and decreased after day 2 (about 330 pmol/mg protein). Immunoblot analysis showed that the mean p36 protein level was low (< 1.6 arbitrary units [AU]) during gestation, increased sharply (to about 4.0 AU) during the first day, and remained stable afterward. Unlike p36, p46 protein was present before birth at values comparable to those postpartum. P46 increased from 3.2 AU at 20 days to 4.6 AU at 21 days of gestation, and decreased transiently after birth. These results show that (1) G6Pase hydrolytic activity before birth can occur without detectable G6P uptake function; (2) the presence of the putative G6P transporter protein is not sufficient to elicit G6P uptake; and (3) full G6Pase activity requires optimal expression of both p36 and p46 proteins. These data are discussed in relation to the function of G6Pase. PMID- 11016905 TI - Kinetics of very-low-density lipoprotein apolipoprotein B-100 in normolipidemic subjects: pooled analysis of stable-isotope studies. AB - To further explore the physiology of very-low-density lipoprotein (VLDL) apolipoprotein B-100 (apoB), we performed a pooled analysis of 21 reports based on the intravenous administration of stable isotope-labeled amino acids in a total of 154 healthy normolipidemic subjects. Prandial status was the most significant independent predictor (P < .001) of the hepatic secretion of apoB, which was higher in the fed state compared with the fasted state (1,819 +/- 188 v 1,046 +/- 61 mg/d, P < .001). In the fed state, apoB secretion increased with age (P = .003) and tended to be higher in men compared with women (P = .0065). The fractional catabolism of VLDL apoB decreased with weight (P = .0038) and was lower in men versus women (8.38 +/- 0.55 v 12.59 +/- 1.65 pools/d, P = .007), as well as patients that were carriers of the E4 allele compared with those who were not carriers of this allele (5.52 +/- 0.49 v 9.58 +/- 0.87 pools/d, P < .001). The VLDL apoB concentration in both the fed and fasted states was dependent on both the rate of hepatic production and fractional clearance of apoB. Plasma cholesterol, triglyceride, low-density lipoprotein (LDL) cholesterol, and high density lipoprotein (HDL) cholesterol concentrations in the fasted state were principally determined by the fractional catabolism of VLDL apoB (P< .005). These findings suggest that under physiologic conditions in healthy individuals, the transport of VLDL apoB in plasma is predominantly determined by age, sex, body weight, apoE genotype, and prandial status. PMID- 11016906 TI - A probing dose of phenylacetate does not affect glucose production and gluconeogenesis in humans. AB - Phenylacetate ingestion has been used to probe Krebs cycle metabolism and to augment waste nitrogen excretion in urea cycle disorders. Phenylalkanoic acids, including phenylacetate, have been proposed as potential therapeutic agents in the treatment of diabetes. They inhibit gluconeogenesis in the liver in vitro and reduce the blood glucose concentration in diabetic rats. The effect of sodium phenylacetate ingestion on blood glucose and the contribution of gluconeogenesis to glucose production have now been studied in 7 type 2 diabetic patients. The study was not designed to test whether the changes in glucose metabolism observed in the rat could be reproduced in humans. After an overnight fast, over a period of 1 hour, 4.8 g phenylacetate was ingested, which is the highest dose used to probe Krebs cycle metabolism. Glucose production was measured by tracer kinetics using [6,6-(2)H2]glucose and gluconeogenesis by the labeling of the hydrogens of blood glucose on (2)H20 ingestion. The concentration of phenylacetate in plasma peaked by 2 hours after its ingestion, and about 40% of the dose was excreted in 5 hours. The plasma glucose concentration and production, and the contribution of gluconeogenesis to glucose production, were unaffected by phenylacetate ingestion at the highest dose used to probe Krebs cycle metabolism. PMID- 11016907 TI - Gln27Glu and Arg16Gly polymorphisms of the beta2-adrenergic receptor gene are not associated with obesity in Japanese men. AB - The beta2-adrenergic receptor (beta2AR), beta3AR, or uncoupling protein 1 (UCP1) may play a pathogenic role in obesity. In Swedish Caucasians, a polymorphism at codon 27 (Gln27Glu) of the beta2AR gene was shown to be associated with obesity, but no such association was shown for a polymorphism of codon 16 (Arg16Gly). Thus, we investigated whether these polymorphisms contribute to obesity in 210 Japanese men. The frequencies of the Gln27Glu and Arg16Gly polymorphisms were 0.05 and 0.48, respectively, and there was no association with obesity. A strong linkage disequilibrium between the Gln27Glu and Arg16Gly polymorphisms was shown, but there was no apparent additive effect on the clinical or metabolic characteristics. Our results suggest that the Gln27Glu and Arg16Gly polymorphisms of the beta2AR gene are not a major contributing factor to obesity in Japanese men. PMID- 11016908 TI - Increased insulin sensitivity and decreased insulin secretion in offspring of insulin-sensitive type 2 diabetic patients. AB - To investigate the early defects of glucose metabolism in insulin-sensitive type 2 diabetes, we performed oral and frequently sampled intravenous glucose tolerance tests (OGTT and FSIGT) with minimal model analysis in 15 offspring of Japanese type 2 diabetics with normal insulin sensitivity (insulin resistance index of homeostasis model assessment [HOMA-R] < 2.0) and in 20 healthy control subjects without a family history of type 2 diabetes. The frequency of impaired glucose tolerance (IGT) was 40% (6 of 15) in the offspring and 0% (0 of 20) in the controls. Fasting plasma glucose (4.8 +/- 0.1 v4.6 +/- 0.1 mmol/L, P = .18) and immunoreactive insulin ([IRI] 29.9 +/- 2.5 v 28.3 +/- 2.5 pmol/L, P = .64) were comparable between the offspring and the controls. The rate of glucose disappearance (KG) was significantly lower in the offspring versus the control group (2.00 +/- 0.22 v 2.60 +/- 0.17 min(-1), P= .03). The insulin sensitivity index (Si) was significantly greater in the offspring versus the controls (2.68 +/- 0.41 v 1.71 +/- 0.17 x 10(-4) min(-1) x pmol/L , P = .02). First-phase insulin secretion (FPI) to intravenous glucose was significantly lower in the offspring versus the control group (886 +/- 110 v 2,296 +/- 267 min x pmol/L, P< .01). Glucose effectiveness (SG) was comparable between the offspring and control groups. The disposition index (Si x FPI) was significantly lower in the offspring versus the controls (2,106 +/- 256 v 3,652 +/- 490 x 10(-4), P = .02). When the offspring were subdivided into 2 groups by glucose tolerance status, both normal glucose tolerance (NGT) offspring and IGT offspring showed a significant decrease in FPI and increase in Si. Thus, although the offspring of insulin-sensitive type 2 diabetics had increased insulin sensitivity, the impairment in insulin secretion was more dominant. Our results suggest that the early metabolic abnormality in insulin-sensitive type 2 diabetes is an insulin secretory dysfunction despite increased insulin sensitivity. PMID- 11016909 TI - Lipoprotein lipase steady-state mRNA levels are lower in human omental versus subcutaneous abdominal adipose tissue. AB - Adipose tissue synthesizes lipoprotein lipase (LPL), which helps in the postprandial clearance of triglyceride-rich lipoproteins. Because visceral adipose tissue is generally accepted as the most important metabolic tissue, we sought to verify whether there are regional differences in the expression of LPL. Samples of adipose tissue from subcutaneous and omental fat deposits were obtained from 20 adults undergoing surgery. Total adipose tissue LPL activity was measured using a conventional radioactive substrate assay. Steady-state levels of LPL mRNA were assessed using the very sensitive RNase protection assay technique with 18S ribosomal RNA as an internal control. A correlation was demonstrated between LPL activity levels in subcutaneous and omental tissue (r = .72; P < .01) and between mRNA levels at both sites (r = .47, P = .04). LPL mRNA levels were significantly lower in omental compared with subcutaneous depots (omental v subcutaneous, 1.7 +/- 0.7 v 2.1 +/- 0.7 arbitrary units [AU] over 18S, P < .05). In paired comparisons, LPL mRNA levels in omental adipose tissue were, on average, 20% +/- 7% (range, -57% to +9.0%) lower than the levels measured in subcutaneous adipose tissue (P < .05). In conclusion, these data suggest that subcutaneous adipose tissue is a reliable surrogate of the expression (activity and mRNA) of LPL in omental adipose tissue, even though omental depots express proportionally less LPL than subcutaneous depots. PMID- 11016910 TI - Interaction on metabolic clearance between A-type and B-type natriuretic peptides in patients with heart failure. AB - A-type and B-type natriuretic peptides (ANP and BNP) are secreted into the systemic circulation via the coronary sinus. Plasma levels of ANP and BNP at the coronary sinus should directly determine the systemic circulating levels. However, the metabolic clearance of these hormones are dependent on similar systems, natriuretic peptide clearance receptor (NPR-C) and neutral endopeptidase 24.11 (NEP), suggesting a possible interaction between ANP and BNP on metabolic clearance. In this study, we examined the interaction on metabolic clearance in patients with heart failure. We obtained blood samples from the coronary sinus and aortic root in 100 patients with heart failure and 28 control subjects. The difference in ANP and BNP levels between the coronary sinus and the aortic root is reflected partly by the metabolic clearance in the pulmonary circulation. In this study, we examined the possible interaction on metabolic clearance between ANP and BNP using a statistical procedure. The ratio of the level of BNP to ANP (BNP/ANP) was significantly higher in the aortic root than in the coronary sinus at any stage of heart failure. We performed multiple regression analysis using ANP and BNP levels at the coronary sinus as independent variables (X1 and X2, respectively) and the ANP level at the aortic root as a dependent variable (Y). The analysis showed that both X1 and X2 were significant variables in the equation. On the other hand, we performed the same analysis using the BNP level at the aortic root as a dependent variable (Y). The analysis showed that only X2 was a significant variable in the equation. This study suggests that (1) the metabolic clearance in the pulmonary circulation is higher for ANP versus BNP and (2) the amount of ANP cleared in the pulmonary circulation depends on the amount of both ANP and BNP secreted from the heart, whereas the amount of BNP cleared in the pulmonary circulation is dependent solely on the amount of BNP secreted from the heart. PMID- 11016912 TI - Validation of a screening questionnaire for androgen deficiency in aging males. AB - It is now well established that testosterone levels decline with age. What has not been established is whether the decline in testosterone is associated with a symptom complex. This study examined whether certain symptoms are more commonly present in males with low bioavailable testosterone (BT) levels. These were used to evaluate a questionnaire for androgen deficiency in aging males (ADAM). The validity of the ADAM questionnaire to screen for low BT was tested in 316 Canadian physicians aged 40 to 62 years. Low BT levels were present in 25% of this population. None had elevated luteinizing hormone (LH) levels. The ADAM questionnaire had 88% sensitivity and 60% specificity. When the questionnaire was administered twice 2 to 4 weeks apart to 10 men, it was determined that the coefficient of variation was 11.5%. In a second study of 34 ADAM-positive patients, 37% of those with clearly normal BT levels demonstrated some evidence of dysphoria. Finally, in 21 patients who were treated with testosterone, improvement on the ADAM questionnaire was demonstrated in 18 (P = .002). These data support the concept of a symptom complex associated with low BT levels in aging males. In addition, the ADAM questionnaire appears to be a reasonable screening questionnaire to detect androgen deficiency in males over 40 years of age. PMID- 11016911 TI - Effects of high-dose simvastatin on adrenal and gonadal steroidogenesis in men with hypercholesterolemia. AB - In view of the role of both the de novo biosynthesis and receptor-mediated uptake of cholesterol for normal steroidogenesis, we evaluated whether extending the therapeutic dose of the hepatic hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase inhibitor, simvastatin, to 80 mg/d would affect adrenal and gonadal steroid synthesis in men with hypercholesterolemia. To evaluate this question, we enrolled men into a multicenter randomized, placebo-controlled study lasting 12 weeks. Men with serum low-density lipoprotein cholesterol (LDL-C) more than 145 mg/dL after 6 weeks of a lipid-lowering diet were randomized to 80 mg simvastatin or placebo. Half of the subjects were asked to undergo a 6-hour infusion of corticotropin (ACTH) to evaluate cortisol synthesis, and the entire cohort received a human chorionic gonadotropin (hCG) stimulation test to assess gonadal hormone secretion using pooled serum samples taken 15 minutes apart. A total of 81 men (age, 45 +/- 11 years; 93% Caucasian) with baseline serum LDL-C of 197 mg/dL (placebo, n = 39) and 184 mg/dL (simvastatin 80 mg, n = 42) completed the study. After 12 weeks, serum LDL-C, triglycerides, and high-density lipoprotein cholesterol (HDL-C) in the simvastatin group changed by -43%, -25%, and 8%, respectively (all P < .001). The basal cortisol level and the peak serum cortisol and area under the curve response to the 6-hour ACTH infusion were comparable between the two treatment groups at baseline and after 12 weeks. The pooled total testosterone level at baseline was 541 and 513 ng/dL in the placebo and simvastatin-treated groups, respectively, which declined to 536 +/- 20.5 ng/dL ( 1.5%) and 474 +/- 30.4 ng/dL (-13.6%, P = .09) after treatment (mean +/- SD). The pooled free testosterone declined by 6.3% in the simvastatin group, versus a 4.9% increase in the placebo group (P = .588), while pooled bioavailable testosterone declined 10.2% in the simvastatin group and increased 1.4% in the placebo group (P = .035). There were no changes in serum gonadotropin levels or sex hormone binding globulin (SHBG). After administration of hCG, there were no differences in the peak total pooled testosterone level before or after 12 weeks of treatment. Simvastatin 80 mg was well tolerated compared with placebo. In conclusion, basal and stimulated cortisol production was unaffected by the use of simvastatin 80 mg versus placebo. As reported with other statins and cholestyramine, there were small declines in the simvastatin-treated group for pooled total, free, and bioavailable testosterone after 12 weeks, although there was no compensatory increase in serum follicle-stimulating hormone (FSH) or luteinizing hormone (LH) levels. PMID- 11016913 TI - Ask the doctor. A hospital near me is advertising electron beam CT scanning of the heart to detect coronary artery disease. I don't have any symptoms of heart disease, but I did undergo this test--paying my own money, I would note! It came back with a fairly high calcium score, and the testing people told me to tell me doctor about it. My doctor says he doesn't know what to do with the information. Do you? PMID- 11016914 TI - By the way, doctor... I'm a 62-year-old woman, and for years I've had an aching chest pain that comes on at unpredictable times and sometimes lasts for hours. My doctor said my stress test results were normal. But I've read that heart disease is harder to diagnose in women than in men. Should I get more tests? PMID- 11016915 TI - Trends in the treatment of anxiety. PMID- 11016916 TI - Assisted suicide and the fluctuating will to live. PMID- 11016917 TI - Smoking and panic. PMID- 11016918 TI - Untreated schizophrenia. PMID- 11016919 TI - Smurf2 is a ubiquitin E3 ligase mediating proteasome-dependent degradation of Smad2 in transforming growth factor-beta signaling. AB - Smads are important intracellular signaling effectors for transforming growth factor-beta (TGF-beta) and related factors. Proper TGF-beta signaling requires precise control of Smad functions. In this study, we have identified a novel HECT class ubiquitin E3 ligase, designated Smurf2, that negatively regulates Smad2 signaling. In both yeast two-hybrid and in vitro binding assays, we found that Smurf2 could interact with receptor-activated Smads (R-Smads), including Smad1, Smad2, and Smad3 but not Smad4. Ectopic expression of Smurf2 was sufficient to reduce the steady-state levels of Smad1 and Smad2 but not Smad3 or Smad4. Significantly, Smurf2 displayed preference to Smad2 as its target for degradation. Furthermore, Smurf2 exhibited higher binding affinity to activated Smad2 upon TGF-beta stimulation. The ability of Smurf2 to promote Smad2 destruction required the HECT catalytic activity of Smurf2 and depended on the proteasome-dependent pathway. Consistent with these results, Smurf2 potently reduced the transcriptional activity of Smad2. These data suggest that a ubiquitin/proteasome-dependent mechanism is important for proper regulation of TGF-beta signaling. PMID- 11016920 TI - Inhibition of c-Myc expression sensitizes hepatocytes to tumor necrosis factor induced apoptosis and necrosis. AB - The typical proliferative response of hepatocytes to tumor necrosis factor (TNF) can be converted to a cytotoxic one by transcriptional arrest. Although NF-kappaB activation is critical for hepatocyte resistance to TNF toxicity, the contribution of other TNF-inducible transcription factors remains unknown. To determine the function of c-Myc in hepatocyte sensitivity to TNF, stable transfectants of the rat hepatocyte cell line RALA255-10G containing sense and antisense c-myc expression vectors were isolated with increased (S-Myc cells) and decreased (AN-Myc cells) c-Myc transcriptional activity. While S-Myc cells proliferated in response to TNF treatment, AN-Myc cells underwent 32% cell death within 6 h. Fluorescent microscopic studies indicated that TNF induced apoptosis and necrosis in AN-Myc cells. Cell death was associated with DNA hypoploidy and poly(ADP-ribose) polymerase cleavage but occurred in the absence of detectable caspase-3, -7, or -8 activation. TNF-induced, AN-Myc cell death was dependent on Fas-associated protein with death domain and partially blocked by caspase inhibitors. AN-Myc cells had decreased levels of NF-kappaB transcriptional activity, but S-Myc cells maintained resistance to TNF despite NF-kappaB inactivation, suggesting that c-Myc and NF-kappaB independently mediate TNF resistance. Thus, in the absence of sufficient c-Myc expression, hepatocytes are sensitized to TNF-induced apoptosis and necrosis. These findings demonstrate that hepatocyte resistance to TNF is regulated by multiple transcriptional activators. PMID- 11016921 TI - A novel nuclear localization signal in human DNA topoisomerase I. AB - DNA topoisomerase (topo) I is a nuclear enzyme that plays an important role in DNA metabolism. Based on conserved nuclear targeting sequences, four classic nuclear localization signals (NLSs) have been proposed at the N terminus of human topo I, but studies with yeast have suggested that only one of them (amino acids (aa) 150-156) is sufficient to direct the enzyme to the nucleus. In this study, we expressed human topo I fused to enhanced green fluorescent protein (EGFP) in mammalian cells and demonstrated that whereas aa 150-156 are sufficient for nuclear localization, the nucleolar localization requires aa 157-199. More importantly, we identified a novel NLS within aa 117-146. In contrast to the classic NLSs that are rich in basic amino acids, the novel NLS identified in this study is rich in acidic amino acids. Furthermore, this novel NLS alone is sufficient to direct not only EGFP into the nucleus but also topo I; and the EGFP.topo I fusion driven by the novel NLS is as active in vivo as the wild-type topo I in response to the topo I inhibitor topotecan. Together, our results suggest that human topo I carries two independent NLSs that have opposite amino acid compositions. PMID- 11016922 TI - Molecular basis of the recruitment of the SH2 domain-containing inositol 5 phosphatases SHIP1 and SHIP2 by fcgamma RIIB. AB - FcgammaRIIB are single-chain low affinity receptors for IgG that negatively regulate immunoreceptor tyrosine-based activation motif-dependent cell activation. They bear one immunoreceptor tyrosine-based inhibition motif (ITIM) that becomes tyrosyl-phosphorylated upon coaggregation of FcgammaRIIB with immunoreceptor tyrosine-based activation motif-bearing receptors and that recruits SH2 domain-containing inositol 5-phosphatases (SHIPs) in vivo. Synthetic FcgammaRIIB ITIM phosphopeptides, however, also bind SH2 domain-containing protein-tyrosine phosphatases (SHPs) in vitro. To identify SHIP-binding sites, we exchanged residues between the FcgammaRIIB ITIM and the N-terminal ITIM of a killer cell Ig-like receptor that does not bind SHIPs. Loss of function and gain of function substitutions identified the Y+2 leucine, in the FcgammaRIIB ITIM, as determining the binding of both SHIP1 and SHIP2, but not the binding of SHP-1 or SHP-2. Conversely, the Y-2 isoleucine that determines the in vitro binding of SHP 1 and SHP-2 affected neither the binding nor the recruitment of SHIP1 or SHIP2. One hydrophobic residue, in the ITIM of FcgammaRIIB therefore determines the affinity for SHIPs. This residue is symmetrical to the hydrophobic residue that determines the affinity of all ITIMs for SHPs. It defines a SHIP-binding site, distinct from a SHP-binding site, that enables FcgammaRIIB to recruit SHIP1 and SHIP2 and that is preferentially used in vivo. PMID- 11016923 TI - Cathepsin B activity regulation. Heparin-like glycosaminogylcans protect human cathepsin B from alkaline pH-induced inactivation. AB - It has been shown that lysosomal cysteine proteinases, specially cathepsin B, has been implicated in a variety of diseases involving tissue remodeling states, such as inflammation, parasite infection, and tumor metastasis, by degradation of extracellular matrix components. Recently, we have shown that heparin and heparan sulfate bind to papain specifically; this interaction induces an increase of its alpha-helix content and stabilizes the enzyme structure even at alkaline pH (Almeida, P. C., Nantes, I. L., Rizzi, C. C. A., Judice, W. A. S., Chagas, J. R., Juliano, L., Nader, H. B., and Tersariol, I. L. S. (1999) J. Biol. Chem. 274, 30433-30438). In the present work, a combination of circular dichroism analysis, affinity chromatography, cathepsin B mutants, and fluorogenic substrate assays were used to characterize the interaction of human cathepsin B with glycosaminoglycans. The nature of the cathepsin B-glycosaminoglycans interaction was sensitive to the charge and type of polysaccharide. Like papain, heparin and heparan sulfate bind cathepsin B specifically, and this interaction reduces the loss of cathepsin B alpha-helix content at alkaline pH. Our data show that the coupling of cathepsin B with heparin or heparan sulfate can potentiate the endopeptidase activity of the cathepsin B, increasing 5-fold the half-life (t(12)) of the enzyme at alkaline pH. Most of these effects are related to the interaction of heparin and heparan sulfate with His(111) residue of the cathepsin B occluding loop. These results strongly suggest that heparan sulfate may be an important binding site for cathepsin B at cell surface, reporting a novel physiological role for heparan sulfate proteoglycans. PMID- 11016924 TI - Analysis of the human lumican gene promoter. AB - The human lumican gene was shown to possess one major transcription start site, resulting in exon 1 of the gene giving rise to the first 74 base pairs (bp) of the 5'-untranslated region. About 1.6 kilobase pairs of upstream promoter sequence were sequenced and analyzed to identify elements responsible for gene expression. No typical TATAA sequence was identified in the vacinity of the transcription start site, but an atypical TATCA sequence residing 41 bp upstream was shown to be necessary for transcription, although it was incapable of supporting transcription by itself. A GC box residing 74 bp upstream of the transcription start site also was essential for the initiation of transcription. Sp3 was identified as the transcriptional activator binding to the GC box. No additional elements that significantly modulated transcription were noted in the promoter sequence analyzed, when using human adult chondrocytes as the cell source for transfection in reporter assays. In contrast, reporter assays carried out in human fetal lung fibroblasts, where lumican expression is deplete, revealed the presence of a repressor element located between 384 and 598 bp upstream of the transcription start site. A GATA-binding site located between bp 386 and -391 was identified as being necessary for repression of transcription. The mouse lumican promoter does not possess an equivalent site, and this may explain why the lumican gene is expressed in fetal murine cartilage but not in fetal human cartilage. PMID- 11016925 TI - Intracellular acidification triggered by mitochondrial-derived hydrogen peroxide is an effector mechanism for drug-induced apoptosis in tumor cells. AB - We recently showed that two photoproducts of merocyanine 540, C2 and C5, triggered cytochrome C release; however, C5 was inefficient in inducing caspase activity and apoptosis in leukemia cells, unlike C2. Here we show that HL60 cells acidified upon exposure to C2 but not C5. The intracellular drop in pH and caspase activation were dependent upon hydrogen peroxide production, and were inhibited by scavengers of hydrogen peroxide. On the contrary, caspase inhibitors did not block hydrogen peroxide production. In turn, increased intracellular hydrogen peroxide concentration was downstream of superoxide anion produced within 2 h of exposure to C2. Inhibitor of NADPH oxidase diphenyleneiodonium neither inhibited superoxide production nor caspase activation triggered by C2. However, exposure of purified mitochondria to C2 resulted in significantly increased superoxide production. Furthermore, cytochrome C release from isolated mitochondria induced by C2 was completely inhibited in the presence of scavengers of hydrogen peroxide. Contrarily, scavenging hydrogen peroxide had no effect on the cyclosporin A-sensitive mitochondrial permeability transition induced by C5. Our data suggest a scenario where drug-induced hydrogen peroxide production induces intracellular acidification and release of cytochrome C, independent of the inner membrane pore, thereby creating an intracellular environment permissive for caspase activation. PMID- 11016926 TI - A potential role for protein kinase C-epsilon in regulating megakaryocytic lineage commitment. AB - Multiple studies have shown that intracellular signal transduction by the protein kinase C (PKC) family participates in the initiation of megakaryocyte differentiation. In this study, multiple approaches addressed the functional contributions by specific PKC isozymes to megakaryocytic lineage commitment of two independent cell lines, K562 and human erythroleukemia (HEL). Pharmacologic profiles of induction and inhibition of megakaryocytic differentiation in both cell lines suggested a role for the calcium-independent novel PKCs, in particular PKC-epsilon. In transfection studies, the isolated variable domain of PKC-epsilon selectively blocked exogenous activation of the megakaryocyte-specific alpha IIb promoter. Constitutively active mutants of PKC-epsilon, but not of other PKC isozymes, cooperated with the transcription factor GATA-1 in the activation of the alpha IIb promoter. The functional cooperation between GATA-1 and PKC-epsilon displayed dependence on cellular milieu, as well as on the promoter context of GATA binding sites. In aggregate, the data suggest that PKC-epsilon specifically participates in megakaryocytic lineage commitment through functional cooperation with GATA-1 in the activation of megakaryocytic promoters. PMID- 11016927 TI - Non-redundant signal transduction of interleukin-6-type cytokines. The adapter protein Shc is specifically recruited to rhe oncostatin M receptor. AB - The common use of the cytokine receptor gp130 has served as an explanation for the extremely redundant biological activities exerted by interleukin (IL)-6-type cytokines. Indeed, hardly any differences in signal transduction initiated by these cytokines are known. In the present study, we demonstrate that oncostatin M (OSM), but not IL-6 or leukemia inhibitory factor, induces tyrosine phosphorylation of the Shc isoforms p52 and p66 and their association with Grb2. Concomitantly, OSM turns out to be a stronger activator of ERK1/2 MAPKs. Shc is recruited to the OSM receptor (OSMR), but not to gp130. Binding involves Tyr(861) of the OSMR, located within a consensus binding sequence for the Shc PTB domain. Moreover, Tyr(861) is essential for activation of ERK1/2 and for full activation of the alpha(2)-macroglobulin promoter, but not for an exclusively STAT responsive promoter. This study therefore provides evidence for qualitative differential signaling mechanisms exerted by IL-6-type cytokines. PMID- 11016928 TI - Retinoblastoma gene promoter directs transgene expression exclusively to the nervous system. AB - In human, germ line mutations in the tumor suppressor retinoblastoma (Rb) predispose individuals to retinoblastoma, whereas somatic inactivation of Rb contributes to the progression of a large spectrum of cancers. In mice, Rb is highly expressed in restricted cell lineages including the neurogenic, myogenic, and hematopoietic systems, and disruption of the gene leads to specific embryonic defects in these tissues. The symmetry between Rb expression and the defects in mutant mice suggest that transcriptional control of Rb during embryogenesis is pivotal for normal development. We have initiated studies to dissect the mechanisms of transcriptional regulation of Rb during development by promoter lacZ transgenic analysis. DNA sequences up to 6 kilobase pairs upstream of the mouse Rb promoter, isolated from two different genomic libraries, directed transgene expression exclusively to the developing nervous system, excluding skeletal muscles and liver. Expression of the transgene in the central and peripheral nervous systems, including the retina, recapitulated the expression of endogenous Rb during embryogenesis. A promoter region spanning approximately 250 base pairs upstream of the transcriptional starting site was sufficient to confer expression in the central and peripheral nervous systems. To determine whether this expression pattern was conserved, we isolated the human Rb 5' genomic region and generated transgenic mice expressing lacZ under control of a 1.6-kilobase pair human Rb promoter. The human Rb promoter lacZ mice also expressed the transgene primarily in the nervous system in several independent lines. Thus, transgene expression directed by both the human and mouse Rb promoters is restricted to a subset of tissues in which Rb is normally expressed during embryogenesis. Our findings demonstrate that regulatory elements directing Rb expression to the nervous system are delineated within a well defined core promoter and are regionally separated from elements, yet to be identified, that are required for expression of Rb in the developing hematopoietic and skeletal muscle systems. PMID- 11016929 TI - Nuclear factors bind to a conserved DNA element that modulates transcription of Anopheles gambiae trypsin genes. AB - The Anopheles gambiae trypsin family consists of seven genes that are transcribed in the gut of female mosquitoes in a temporal coordinated and mutually exclusive manner, suggesting the involvement of a complex transcription regulatory mechanism. We identified a highly conserved 12-nucleotide motif present in all A. gambiae and Anopheles stephensi trypsin promoters. We investigated the role of this putative trypsin regulatory element (PTRE) in controlling the transcription of the trypsin genes. Gel shift experiments demonstrated that nuclear proteins of A. gambiae cell lines formed two distinct complexes with probes encompassing the PTRE sequence. Mapping of the binding sites revealed that one of the complex has the specificity of a GATA transcription factor. Promoter constructs containing mutations in the PTRE sequence that selectively abolished the binding of either one or both complexes exerted opposite effects on the transcriptional activity of trypsin promoters in A. gambiae and Aedes aegypti cell lines. In addition, the expression of a novel GATA gene was highly enriched in A. gambiae guts. Taken together our data prove that factors binding to the PTRE region are key regulatory elements possibly involved in the blood meal-induced repression and activation of transcription in early and late trypsin genes. PMID- 11016930 TI - The N-terminal end of nebulin interacts with tropomodulin at the pointed ends of the thin filaments. AB - Strict regulation of actin thin filament length is critical for the proper functioning of sarcomeres, the basic contractile units of myofibrils. It has been hypothesized that a molecular template works with actin filament capping proteins to regulate thin filament lengths. Nebulin is a giant protein ( approximately 800 kDa) in skeletal muscle that has been proposed to act as a molecular ruler to specify the thin filament lengths characteristic of different muscles. Tropomodulin (Tmod), a pointed end thin filament capping protein, has been shown to maintain the final length of the thin filaments. Immunofluorescence microscopy revealed that the N-terminal end of nebulin colocalizes with Tmod at the pointed ends of thin filaments. The three extreme N-terminal modules (M1-M2-M3) of nebulin bind specifically to Tmod as demonstrated by blot overlay, bead binding, and solid phase binding assays. These data demonstrate that the N terminus of the nebulin molecule extends to the extreme end of the thin filament and also establish a novel biochemical function for this end. Two Tmod isoforms, erythrocyte Tmod (E-Tmod), expressed in embryonic and slow skeletal muscle, and skeletal Tmod (Sk-Tmod), expressed late in fast skeletal muscle differentiation, bind on overlapping sites to recombinant N-terminal nebulin fragments. Sk-Tmod binds nebulin with higher affinity than E-Tmod does, suggesting that the Tmod/nebulin interaction exhibits isoform specificity. These data provide evidence that Tmod and nebulin may work together as a linked mechanism to control thin filament lengths in skeletal muscle. PMID- 11016931 TI - The formation of a flexible DNA-binding protein chain is required for efficient DNA unwinding and adenovirus DNA chain elongation. AB - The adenovirus DNA-binding protein (DBP) binds cooperatively to single-stranded DNA (ssDNA) and stimulates both initiation and elongation of DNA replication. DBP consists of a globular core domain and a C-terminal arm that hooks onto a neighboring DBP molecule to form a stable protein chain with the DNA bound to the internal surface of the chain. This multimerization is the driving force for ATP independent DNA unwinding by DBP during elongation. As shown by x-ray diffraction of different crystal forms of the C-terminal domain, the C-terminal arm can adopt different conformations, leading to flexibility in the protein chain. This flexibility is a function of the hinge region, the part of the protein joining the C-terminal arm to the protein core. To investigate the function of the flexibility, proline residues were introduced in the hinge region, and the proteins were purified to homogeneity after baculovirus expression. The mutant proteins were still able to bind ss- and double-stranded DNA with approximately the same affinity as wild type, and the binding to ssDNA was found to be cooperative. All mutant proteins were able to stimulate the initiation of DNA replication to near wild type levels. However, the proline mutants could not support elongation of DNA replication efficiently. Even the elongation up to 26 nucleotides was severely impaired. This defect was also seen when DNA unwinding was studied. Binding studies of DBP to homo-oligonucleotides showed an inability of the proline mutants to bind to poly(dA)(40), indicating an inability to adapt to specific DNA conformations. Our data suggest that the flexibility of the protein chain formed by DBP is important in binding and unwinding of DNA during adenovirus DNA replication. A model explaining the need for flexibility of the C terminal arm is proposed. PMID- 11016932 TI - Identification of a major heparin-binding site in kallistatin. AB - Kallistatin is a heparin-binding serine proteinase inhibitor (serpin), which specifically inhibits human tissue kallikrein by forming a covalent complex. The inhibitory activity of kallistatin is blocked upon its binding to heparin. In this study we attempted to locate the heparin-binding site of kallistatin using synthetic peptides derived from its surface regions and by site-directed mutagenesis of basic residues in these surface regions. Two synthetic peptides, containing clusters of positive-charged residues, one derived from the F helix and the other from the region encompassing the H helix and C2 sheet of kallistatin, were used to assess their heparin binding activity. Competition assay analysis showed that the peptide derived from the H helix and C2 sheet displayed higher and specific heparin binding activity. The basic residues in both regions were substituted to generate three kallistatin double mutants K187A/K188A (mutations in the F helix) and K307A/R308A and K312A/K313A (mutations in the region between the H helix and C2 sheet), using a kallistatin P1Arg variant as a scaffold. Analysis of these mutants by heparin-affinity chromatography showed that the heparin binding capacity of the variant K187A/K188A was not altered, whereas the binding capacity of K307A/R308A and K312A/K313A mutants was markedly reduced. Titration analysis with heparin showed that the K312A/K313A mutant has the highest dissociation constant. Like kallistatin, the binding activity of K187A/K188A to tissue kallikrein was blocked by heparin, whereas K307A/R308A and K312A/K313A retained significant binding and inhibitory activities in the presence of heparin. These results indicate that the basic residues, particularly Lys(312)-Lys(313), in the region between the H helix and C2 sheet of kallistatin, comprise a major heparin-binding site responsible for its heparin-suppressed tissue kallikrein binding. PMID- 11016933 TI - Zebrafish acetylcholinesterase is encoded by a single gene localized on linkage group 7. Gene structure and polymorphism; molecular forms and expression pattern during development. AB - We cloned and sequenced the acetylcholinesterase gene and cDNA of zebrafish, Danio rerio. We found a single gene (ache) located on linkage group LG7. The relative organization of ache, eng2, and shh genes is conserved between zebrafish and mammals and defines a synteny. Restriction fragment length polymorphism analysis was allowed to identify several allelic variations. We also identified two transposable elements in non-coding regions of the gene. Compared with other vertebrate acetylcholinesterase genes, ache gene contains no alternative splicing at 5' or 3' ends where only a T exon is present. The translated sequence is 60 80% identical to acetylcholinesterases of the vertebrates and exhibits an extra loop specific to teleosts. Analysis of molecular forms showed a transition, at the time of hatching, from the globular G4 form to asymmetric A12 form that becomes prominent in adults. In situ hybridization and enzymatic activity detection on whole embryos confirmed early expression of the acetylcholinesterase gene in nervous and muscular tissues. We found no butyrylcholinesterase gene or activity in Danio. These findings make zebrafish a promising model to study function of acetylcholinesterase during development and regulation of molecular forms assembly in vivo. PMID- 11016934 TI - Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins. AB - Poly(ADP-ribose) is formed in possibly all multicellular organisms by a familiy of poly(ADP-ribose) polymerases (PARPs). PARP-1, the best understood and until recently the only known member of this family, is a DNA damage signal protein catalyzing its automodification with multiple, variably sized ADP-ribose polymers that may contain up to 200 residues and several branching points. Through these polymers, PARP-1 can interact noncovalently with other proteins and alter their functions. Here we report the discovery of a poly(ADP-ribose)-binding sequence motif in several important DNA damage checkpoint proteins. The 20-amino acid motif contains two conserved regions: (i) a cluster rich in basic amino acids and (ii) a pattern of hydrophobic amino acids interspersed with basic residues. Using a combination of alanine scanning, polymer blot analysis, and photoaffinity labeling, we have identified poly(ADP-ribose)-binding sites in the following proteins: p53, p21(CIP1/WAF1), xeroderma pigmentosum group A complementing protein, MSH6, DNA ligase III, XRCC1, DNA polymerase epsilon, DNA-PK(CS), Ku70, NF-kappaB, inducible nitric-oxide synthase, caspase-activated DNase, and telomerase. The poly(ADP-ribose)-binding motif was found to overlap with five important functional domains responsible for (i) protein-protein interactions, (ii) DNA binding, (iii) nuclear localization, (iv) nuclear export, and (v) protein degradation. Thus, PARPs may target specific signal network proteins via poly(ADP-ribose) and regulate their domain functions. PMID- 11016935 TI - Altered cytokine production in mice lacking P2X(7) receptors. AB - The P2X(7) receptor (P2X(7)R) is an ATP-gated ion channel expressed by monocytes and macrophages. To directly address the role of this receptor in interleukin (IL)-1 beta post-translational processing, we have generated a P2X(7)R-deficient mouse line. P2X(7)R(-/-) macrophages respond to lipopolysaccharide and produce levels of cyclooxygenase-2 and pro-IL-1 beta comparable with those generated by wild-type cells. In response to ATP, however, pro-IL-1 beta produced by the P2X(7)R(-/-) cells is not externalized or activated by caspase-1. Nigericin, an alternate secretion stimulus, promotes release of 17-kDa IL-1 beta from P2X(7)R( /-) macrophages. In response to in vivo lipopolysaccharide injection, both wild type and P2X(7)R(-/-) animals display increases in peritoneal lavage IL-6 levels but no detectable IL-1. Subsequent ATP injection to wild-type animals promotes an increase in IL-1, which in turn leads to additional IL-6 production; similar increases did not occur in ATP-treated, LPS-primed P2X(7)R(-/-) animals. Absence of the P2X(7)R thus leads to an inability of peritoneal macrophages to release IL 1 in response to ATP. As a result of the IL-1 deficiency, in vivo cytokine signaling cascades are impaired in P2X(7)R-deficient animals. Together these results demonstrate that P2X(7)R activation can provide a signal that leads to maturation and release of IL-1 beta and initiation of a cytokine cascade. PMID- 11016936 TI - Nucleocytoplasmic shuttling of Bruton's tyrosine kinase. AB - Bruton's tyrosine kinase (Btk), a nonreceptor cytoplasmic tyrosine kinase belonging to the Tec family of kinases, has been shown to be critical for B cell proliferation, differentiation, and signaling. Loss-of-function mutations in the Btk gene lead to X-linked agammaglobulinemia (XLA), a primary immunodeficiency in humans, and the less severe condition xid in mice. Although Btk is mainly localized in the cytoplasm under steady state conditions, it translocates to the plasma membrane upon growth factor stimulation and cross-linking of the B cell receptor. Nevertheless, in ectopically as well as endogenously Btk-expressing cells, it can also translocate to the nucleus. Deletion of the pleckstrin homology (PH) domain (DeltaPH1) leads, however, to an even redistribution of Btk within the nucleus and cytoplasm in the majority of transfected cells. In contrast, an SH3-deleted (DeltaSH3) mutant of Btk has been found to be predominantly nuclear. We also demonstrate that the nuclear accumulation of DeltaPH1 is dependent on Src expression. This nucleocytoplasmic shuttling is sensitive to the exportin 1/CRM1-inactivating drug, leptomycin B, indicating that Btk utilizes functional nuclear export signals. In addition, while the DeltaPH1 mutant of Btk was found to be active and tyrosine-phosphorylated in vivo, DeltaSH3 displayed decreased autokinase activity and was not phosphorylated. Our findings indicate that the nucleocytoplasmic shuttling of Btk has implications regarding potential targets inside the nucleus, which may be critical in gene regulation during B cell development and differentiation. PMID- 11016937 TI - Physical characterization of the procollagen module of human thrombospondin 1 expressed in insect cells. AB - Thrombospondin 1 (TSP1) is a homotrimeric glycoprotein composed of 150-kDa subunits connected by disulfide bridges. The procollagen module of thrombospondin 1 has been implicated in antiangiogenic activity. Procollagen modules are found in a number of extracellular proteins and are identifiable by 10 cysteines with characteristic spacing. We expressed and studied the procollagen module (C) of human TSP1, both by itself and in the context of the adjoining oligomerization sequence (o) and N-terminal module (N). The coding sequences were introduced into baculoviruses along with an N-terminal signal sequence and C-terminal polyhistidine tag. Proteins were purified from conditioned medium of infected insect cells by nickel-chelate chromatography. NoC is a disulfide bonded trimer and cleaves readily at a site of preferential proteolysis to yield monomeric N and trimeric oC. These are known properties of full-length TSP1. Mass spectroscopy indicated that C is N-glycosylated, and all 10 cysteine residues of C are in disulfides. By equilibrium ultracentrifugation, C is a monomer in physiological salt solution. Circular dichroism, intrinsic fluorescence, and differential scanning calorimetry experiments suggest that the stability of C is determined by the disulfides. The two tryptophans of C are in a polar, exposed environment as assessed by iodide fluorescence quenching and solvent perturbation. The oC far UV circular dichroism spectrum could be modeled as the sum of C and a coiled-coil oligomerization domain. The results indicate that the recombinant C folds autonomously into its native structure, and trimerization of the modules in TSP1 does not perturb their structures. PMID- 11016938 TI - A functional comparison of BRCA1 C-terminal domains in transcription activation and chromatin remodeling. AB - The BRCA1 C-terminal (BRCT) domain is present in a number of proteins that are involved in various aspects of chromosomal events. The BRCT domain of BRCA1 is important for its function in DNA repair and transcriptional activation. When tethered to chromosomal DNA, this region of BRCA1 is capable of inducing changes in chromatin structure. Despite the sequence homology and functional proximity shared by the BRCT-containing proteins, it is not clear whether different BRCT domains confer a common biochemical activity. Much less is known about the functional significance of the characteristic amino acid residues in the BRCT motif. Here, we show that chromatin remodeling and transcription activation is not a common feature of BRCT domains. However, the BRCT domain of the multi functional repressor-activator protein 1 (RAP1) can activate transcription and remodel chromatin in a manner similar to that shown for the BRCA1 BRCT domain. Most of the conserved amino acid residues in the second BRCA1 BRCT domain are essential for its function in transcriptional activation. In contrast, mutations of many analogous amino acid residues in the RAP1 BRCT domain greatly elevate the transcriptional activity. These data indicate that the conserved residues in these two BRCT domains may play different roles in transcriptional activation. PMID- 11016939 TI - p21-activated kinase 1 activates the nuclear factor kappa B (NF-kappa B)-inducing kinase-Ikappa B kinases NF-kappa B pathway and proinflammatory cytokines in Helicobacter pylori infection. AB - Helicobacter pylori, the causative agent of several human gastric diseases, induces activation of the immediate early response transcription factor nuclear factor kappaB (NF-kappaB), which subsequently triggers release of proinflammatory cytokines in colonized epithelial cells. Here we report that in H. pylori infection p21-activated kinase 1 (PAK1) activates NF-kappaB. Activated PAK1 associates with NF-kappaB-inducing kinase, which upon activation directs the activity of IkappaB kinases to IkappaBalpha. Our results indicate that in epithelial cells PAK1 participates in a unique pathway that links H. pylori dependent effector molecules to the activation of NF-kappaB and the induction of the innate immune response. PMID- 11016940 TI - Calcineurin blockade prevents cardiac mitogen-activated protein kinase activation and hypertrophy in renovascular hypertension. AB - Chronic stimulation of the renin-angiotensin system induces an elevation of blood pressure and the development of cardiac hypertrophy via the actions of its effector, angiotensin II. In cardiomyocytes, mitogen-activated protein kinases as well as protein kinase C isoforms have been shown to be important in the transduction of trophic signals. The Ca(2+)/calmodulin-dependent phosphatase calcineurin has also been suggested to play a role in cardiac growth. In the present report, we investigate possible cross-talks between calcineurin, protein kinase C, and mitogen-activated protein kinase pathways in controlling angiotensin II-induced hypertrophy. Angiotensin II-stimulated cardiomyocytes and mice with angiotensin II-dependent renovascular hypertension were treated with the calcineurin inhibitor cyclosporin A. Calcineurin, protein kinase C, and mitogen-activated protein kinase activations were determined. We show that cyclosporin A blocks angiotensin II-induced mitogen-activated protein kinase activation in cultured primary cardiomyocytes and in the heart of hypertensive mice. Cyclosporin A also inhibits specific protein kinase C isoforms. In vivo, cyclosporin A prevents the development of cardiac hypertrophy, and this effect appears to be independent of hemodynamic changes. These data suggest cross-talks between the calcineurin pathway, the protein kinase C, and the mitogen-activated protein kinase signaling cascades in transducing angiotensin II-mediated stimuli in cardiomyocytes and could provide the basis for an integrated model of cardiac hypertrophy. PMID- 11016941 TI - The channel-forming protein proaerolysin remains a dimer at low concentrations in solution. AB - Proaerolysin, the proform of the channel-forming protein aerolysin, is secreted as a dimer by Aeromonas sp. The protein also exists as a dimer in the crystal, as well as in solution, at least at concentrations in the region of 500 microg/ml. Recently it has been argued that proaerolysin becomes monomeric at concentrations below 100 microg/ml and that only the monomeric form of the protoxin can bind to cell surface receptors (Fivaz, M., Velluz, M.-C., and van der Goot, F. G. (1999) J. Biol. Chem. 274, 37705-37708). Here we show, using non-denaturing polyacrylamide electrophoresis, chemical cross-linking, and analytical ultracentrifugation, that proaerolysin remains dimeric at the lowest concentrations of the protein that we measured (less than 5 microg/ml) and that the dimeric protoxin is quite capable of receptor binding. PMID- 11016942 TI - Mechanism of activation of ERK2 by dual phosphorylation. AB - The mitogen-activated protein (MAP) kinases are characterized by their requirement for dual phosphorylation at a conserved threonine and tyrosine residue for catalytic activation. The structural consequences of dual phosphorylation in the MAP kinase ERK2 (extracellular signal-regulated kinase 2) include active site closure, alignment of key catalytic residues that interact with ATP, and remodeling of the activation loop. In this study, we report the specific effects of dual phosphorylation on the individual catalytic reaction steps in ERK2. Dual phosphorylation leads to an increase in overall catalytic efficiency and turnover rate of approximately 600,000- and 50,000-fold, respectively. Solvent viscosometric studies reveal moderate decreases in the equilibrium dissociation constants (K(d)) for both ATP and myelin basic protein. However, the majority of the overall rate enhancement is due to an increase in the rate of the phosphoryl group transfer step by approximately 60,000-fold. By comparison, the rate of the same step in the ATPase reaction is enhanced only 2000-fold. This suggests that optimizing the position of the invariant residues Lys(52) and Glu(69), which stabilize the phosphates of ATP, accounts for only part of the enhanced rate of phosphoryl group transfer in the kinase reaction. Thus, significant stabilization of the protein phosphoacceptor group must also occur. Our results demonstrate similarities between the activation mechanisms of ERK2 and the cell cycle control enzyme, Cdk2 (cyclin-dependent kinase 2). Rather than dual phosphorylation, however, activation of the latter is controlled by cyclin binding followed by phosphorylation at Thr(160). PMID- 11016943 TI - Regulation of the DPP1-encoded diacylglycerol pyrophosphate (DGPP) phosphatase by inositol and growth phase. Inhibition of DGPP phosphatase activity by CDP diacylglyceron and activation of phosphatidylserine synthase activity by DGPP. AB - The regulation of the Saccharomyces cerevisiae DPP1-encoded diacylglycerol pyrophosphate (DGPP) phosphatase by inositol supplementation and growth phase was examined. Addition of inositol to the growth medium resulted in a dose-dependent increase in the level of DGPP phosphatase activity in both exponential and stationary phase cells. Activity was greater in stationary phase cells when compared with exponential phase cells, and the inositol- and growth phase dependent regulations of DGPP phosphatase were additive. Analyses of DGPP phosphatase mRNA and protein levels, and expression of beta-galactosidase activity driven by a P(DPP1)-lacZ reporter gene, indicated that a transcriptional mechanism was responsible for this regulation. Regulation of DGPP phosphatase by inositol and growth phase occurred in a manner that was opposite that of many phospholipid biosynthetic enzymes. Regulation of DGPP phosphatase expression by inositol supplementation, but not growth phase, was altered in opi1Delta, ino2Delta, and ino4Delta phospholipid synthesis regulatory mutants. CDP diacylglycerol, a phospholipid pathway intermediate used for the synthesis of phosphatidylserine and phosphatidylinositol, inhibited DGPP phosphatase activity by a mixed mechanism that caused an increase in K(m) and a decrease in V(max). DGPP stimulated the activity of pure phosphatidylserine synthase by a mechanism that increased the affinity of the enzyme for its substrate CDP-diacylglycerol. Phospholipid composition analysis of a dpp1Delta mutant showed that DGPP phosphatase played a role in the regulation of phospholipid metabolism by inositol, as well as regulating the cellular levels of phosphatidylinositol. PMID- 11016944 TI - Reexamination of the role of the leucine/isoleucine zipper residues of phospholamban in inhibition of the Ca2+ pump of cardiac sarcoplasmic reticulum. AB - Phospholamban is a small phosphoprotein inhibitor of the Ca(2+)-pump in cardiac sarcoplasmic reticulum, which shows a distinct oligomeric distribution between monomers and homopentamers that are stabilized through Leu/Ile zipper interactions. A two-faced model of phospholamban inhibition of the Ca(2+)-pump was proposed, in which the Leu/Ile zipper residues located on one face of the transmembrane alpha-helix regulate the pentamer to monomer equilibrium, whereas residues on the other face of the helix bind to and inhibit the pump. Here we tested this two-faced model of phospholamban action by analyzing the functional effects of a new series of Leu/Ile zipper mutants. Pentameric stabilities of the mutants were quantified at different SDS concentrations. We show that several phospholamban mutants with hydrophobic amino acid substitutions at the Leu/Ile zipper region retain the ability to form pentamers but at the same time give the same or even stronger (i.e. L37I-PLB) inhibition of the Ca(2+)-pump than do mutants that are more completely monomeric. Steric constraints prevent the Leu/Ile zipper residues sequestered in the interior of the phospholamban pentamer from binding to the Ca(2+)-pump, leading to the conclusion that the zipper residues access the pump from the phospholamban monomer, which is the active inhibitory species. A modified model of phospholamban transmembrane domain action is proposed, in which the membrane span of the phospholamban monomer maintains contacts with the Ca(2+)-pump around most of its circumference, including residues located in the Leu/Ile zipper region. PMID- 11016945 TI - Sodium channel activity in leukemia cells is directly controlled by actin polymerization. AB - The actin cytoskeleton has been shown to be involved in the regulation of sodium selective channels in non-excitable cells. However, the molecular mechanisms underlying the changes in channel function remain to be defined. In the present work, inside-out patch experiments were employed to elucidate the role of submembranous actin dynamics in the control of sodium channels in human myeloid leukemia K562 cells. We found that the application of cytochalasin D to the cytoplasmic surface of membrane fragments resulted in activation of non-voltage gated sodium channels of 12 picosiemens conductance. Similar effects could be evoked by addition of the actin-severing protein gelsolin to the bath cytosol like solution containing 1 microm [Ca(2+)](i). The sodium channel activity induced by disassembly of submembranous microfilaments with cytochalasin D or gelsolin could be abolished by intact actin added to the bath cytosol-like solution in the presence of 1 mm MgCl(2) to induce actin polymerization. In the absence of MgCl(2), addition of intact actin did not abolish the channel activity. Moreover, the sodium currents were unaffected by heat-inactivated actin or by actin whose polymerizability was strongly reduced by cleavage with specific Escherichia coli A2 protease ECP32. Thus, the inhibitory effect of actin on channel activity was observed only under conditions promoting rapid polymerization. Taken together, our data show that sodium channels are directly controlled by dynamic assembly and disassembly of submembranous F-actin. PMID- 11016946 TI - K+ and ionic strength directly influence the autophosphorylation activity of the putative turgor sensor KdpD of Escherichia coli. AB - The membrane-bound histidine kinase KdpD is a putative turgor sensor that regulates, together with the response regulator KdpE, the expression of the kdpFABC operon coding for the high affinity K(+)-uptake system KdpFABC of Escherichia coli. To elucidate the nature of the primary stimulus for KdpD, we developed an in vitro assay based on right-side-out membrane vesicles. Conditions were varied inside and outside of the vesicles, and KdpD autophosphorylation activity was tested. It was shown that an increase of the ionic strength inside the vesicles was accompanied by an increase of the autophosphorylation activity of KdpD with ATP. However, K(+) at concentrations higher than 1 mm inhibited KdpD autophosphorylation activity. This K(+)-specific effect was not observed with KdpD-Arg-511 --> Gln, a KdpD derivative, which causes K(+)-independent kdpFABC expression. When the osmolality outside the vesicles was increased, autophosphorylation activity of KdpD was stimulated, whereby salts were more effective than sugars. Treatment of the vesicles with amphipathic compounds did not affect KdpD autophosphorylation activity. Based on these results it is proposed that changes of intracellular parameters elicited by K(+) limitation or osmotic upshock directly influence KdpD autophosphorylation activity, whereby K(+) has an inhibitory and ionic strength a stimulatory effect. PMID- 11016947 TI - Activation of atypical protein kinase C zeta by caspase processing and degradation by the ubiquitin-proteasome system. AB - Atypical protein kinase C zeta (PKCzeta) is known to transduce signals that influence cell proliferation and survival. Here we show that recombinant human caspases can process PKCzeta at three sites in the hinge region between the regulatory and catalytic domains. Caspase-3, -6, -7, and -8 chiefly cleaved human PKCzeta at EETD downward arrowG, and caspase-3 and -7 also cleaved PKCzeta at DGMD downward arrowG and DSED downward arrowL, respectively. Processing of PKCzeta expressed in transfected cells occurred chiefly at EETD downward arrowG and DGMD downward arrowG and produced carboxyl-terminal polypeptides that contained the catalytic domain. Epitope-tagged PKCzeta that lacked the regulatory domain was catalytically active following expression in HeLa cells. Induction of apoptosis in HeLa cells by tumor necrosis factor alpha plus cycloheximide evoked the conversion of full-length epitope-tagged PKCzeta to two catalytic domain polypeptides and increased PKCzeta activity. A caspase inhibitor, zVAD-fmk, prevented epitope-tagged PKCzeta processing and activation following the induction of apoptosis. Induction of apoptosis in rat parotid C5 cells produced catalytic domain polypeptides of endogenous PKCzeta and increased PKCzeta activity. Caspase inhibitors prevented the increase in PKCzeta activity and production of the catalytic domain polypeptides. Treatment with lactacystin, a selective inhibitor of the proteasome, caused polyubiquitin-PKCzeta conjugates to accumulate in cells transfected with the catalytic domain or full-length PKCzeta, or with a PKCzeta mutant that was resistant to caspase processing. We conclude that caspases process PKCzeta to carboxyl-terminal fragments that are catalytically active and that are degraded by the ubiquitin-proteasome pathway. PMID- 11016948 TI - Powerful anticonvulsant action of IL-1 receptor antagonist on intracerebral injection and astrocytic overexpression in mice. AB - IL-1beta and its endogenous receptor antagonist (IL-1Ra) are rapidly induced by seizures in the rodent hippocampus. Exogenously applied IL-1beta prolongs seizures in an IL-1R type I-mediated manner. This effect depends on N-methyl-d aspartate receptor activation. We report here that intrahippocampal application of recombinant IL-1Ra or its selective endogenous overexpression in astrocytes under the control of glial acidic fibrillary protein promoter potently inhibits motor and electroencephalographic seizures induced by bicuculline methiodide in mice. Accordingly, transgenic mice show a reduced seizure-related c-fos mRNA expression in various forebrain areas compared with their wild-type littermates. Recombinant IL-1Ra was ineffective in mice deficient in IL-1R type I, having per se a delayed onset to generalized convulsions. These results demonstrate that IL 1Ra mediates potent anticonvulsant effects acting on IL-1R type I and suggest that the balance between brain IL-1beta and IL-1Ra represents a crucial mechanism to control seizure generalization. PMID- 11016949 TI - Chemical defense: aquatic beetle (Dineutes hornii) vs. fish (Micropterus salmoides). AB - Captive largemouth bass (Micropterus salmoides) reject the gyrinid beetle, Dineutes hornii. They also reject edible items (mealworms) treated by topical addition of the norsesquiterpene gyrinidal, the principal component of the defensive secretion of the beetle. The bass' oral tolerance of gyrinidal varies broadly as a function of the gyrinidal dosage and the state of satiation of the fish. When taking a D. hornii or a gyrinidal-treated mealworm in the mouth, the fish subjects the item to an intensive oral flushing behavior, seemingly intended to rid the item of gyrinidal. The duration of oral flushing is itself a function of the gyrinidal dosage and the state of satiation of the bass. To counter oral flushing, D. hornii emits its secretion as a slow trickle. Duration of emission is slightly longer (1.5 min) than the time (1.3 min) invested by the bass in flushing a D. hornii before rejecting the beetle. We postulate that flush resistance may be a general feature of defensive chemical delivery systems in aquatic prey, given that oral flushing may be a common strategy of fish. PMID- 11016950 TI - Genome sequence of Halobacterium species NRC-1. AB - We report the complete sequence of an extreme halophile, Halobacterium sp. NRC-1, harboring a dynamic 2,571,010-bp genome containing 91 insertion sequences representing 12 families and organized into a large chromosome and 2 related minichromosomes. The Halobacterium NRC-1 genome codes for 2,630 predicted proteins, 36% of which are unrelated to any previously reported. Analysis of the genome sequence shows the presence of pathways for uptake and utilization of amino acids, active sodium-proton antiporter and potassium uptake systems, sophisticated photosensory and signal transduction pathways, and DNA replication, transcription, and translation systems resembling more complex eukaryotic organisms. Whole proteome comparisons show the definite archaeal nature of this halophile with additional similarities to the Gram-positive Bacillus subtilis and other bacteria. The ease of culturing Halobacterium and the availability of methods for its genetic manipulation in the laboratory, including construction of gene knockouts and replacements, indicate this halophile can serve as an excellent model system among the archaea. PMID- 11016951 TI - Increase of androgen-induced cell death and androgen receptor transactivation by BRCA1 in prostate cancer cells. AB - Although mutations of the breast cancer susceptibility gene 1 (BRCA1) may play important roles in breast and prostate cancers, the detailed mechanism linking the functions of BRCA1 to these two hormone-related tumors remains to be elucidated. Here, we report that BRCA1 interacts with androgen receptor (AR) and enhances AR target genes, such as p21((WAF1/CIP1)), that may result in the increase of androgen-induced cell death in prostate cancer cells. The BRCA1 enhanced AR transactivation can be further induced synergistically with AR coregulators, such as CBP, ARA55, and ARA70. Together, these data suggest that the BRCA1 may function as an AR coregulator and play positive roles in androgen induced cell death in prostate cancer cells and other androgen/AR target organs. PMID- 11016952 TI - High-affinity ouabain binding by a chimeric gastric H+,K+-ATPase containing transmembrane hairpins M3-M4 and M5-M6 of the alpha 1-subunit of rat Na+,K+ ATPase. AB - Na(+),K(+)-ATPase and gastric H(+),K(+)-ATPase are two related enzymes that are responsible for active cation transport. Na(+), K(+)-ATPase activity is inhibited specifically by ouabain, whereas H(+),K(+)-ATPase is insensitive to this drug. Because it is not known which parts of the catalytic subunit of Na(+),K(+)-ATPase are responsible for ouabain binding, we prepared chimeras in which small parts of the alpha-subunit of H(+),K(+)-ATPase were replaced by their counterparts of the alpha(1)-subunit of rat Na(+),K(+)-ATPase. A chimeric enzyme in which transmembrane segments 5 and 6 of H(+), K(+)-ATPase were replaced by those of Na(+),K(+)-ATPase could form a phosphorylated intermediate, but hardly showed a K(+)-stimulated dephosphorylation reaction. When transmembrane segments 3 and 4 of Na(+),K(+)-ATPase were also included in this chimeric ATPase, K(+)-stimulated dephosphorylation became apparent. This suggests that there is a direct interaction between the hairpins M3-M4 and M5-M6. Remarkably, this chimeric enzyme, HN34/56, had obtained a high-affinity ouabain-binding site, whereas the rat Na(+), K(+)-ATPase, from which the hairpins originate, has a low affinity for ouabain. The low affinity of the rat Na(+),K(+)-ATPase previously had been attributed to the presence of two charged amino acids in the extracellular domain between M1 and M2. In the HN34/56 chimera, the M1/M2 loop, however, originates from H(+),K(+)-ATPase, which has two polar uncharged amino acids on this position. Placement of two charged amino acids in the M1/M2 loop of chimera HN34/56 results in a decreased ouabain affinity. This indicates that although the M1/M2 loop affects the ouabain affinity, binding occurs when the M3/M4 and M5/M6 hairpins of Na(+),K(+)-ATPase are present. PMID- 11016953 TI - HIV-1 integrase inhibitors that compete with the target DNA substrate define a unique strand transfer conformation for integrase. AB - Diketo acids such as L-731,988 are potent inhibitors of HIV-1 integrase that inhibit integration and viral replication in cells. These compounds exhibit the unique ability to inhibit the strand transfer activity of integrase in the absence of an effect on 3' end processing. To understand the reasons for this distinct inhibitory profile, we developed a scintillation proximity assay that permits analysis of radiolabeled inhibitor binding and integrase function. High affinity binding of L-731,988 is shown to require the assembly of a specific complex on the HIV-1 long terminal repeat. The interaction of L-731,988 with the complex and the efficacy of L-731, 988 in strand transfer can be abrogated by the interaction with target substrates, suggesting competition between the inhibitor and the target DNA. The L-731,988 binding site and that of the target substrate are thus distinct from that of the donor substrate and are defined by a conformation of integrase that is only adopted after assembly with the viral end. These results elucidate the basis for diketo acid inhibition of strand transfer and have implications for integrase-directed HIV-1 drug discovery efforts. PMID- 11016954 TI - AGO1, QDE-2, and RDE-1 are related proteins required for post-transcriptional gene silencing in plants, quelling in fungi, and RNA interference in animals. AB - Introduction of transgene DNA may lead to specific degradation of RNAs that are homologous to the transgene transcribed sequence through phenomena named post transcriptional gene silencing (PTGS) in plants, quelling in fungi, and RNA interference (RNAi) in animals. It was shown previously that PTGS, quelling, and RNAi require a set of related proteins (SGS2, QDE-1, and EGO-1, respectively). Here we report the isolation of Arabidopsis mutants impaired in PTGS which are affected at the Argonaute1 (AGO1) locus. AGO1 is similar to QDE-2 required for quelling and RDE-1 required for RNAi. Sequencing of ago1 mutants revealed one amino acid essential for PTGS that is also present in QDE-2 and RDE-1 in a highly conserved motif. Taken together, these results confirm the hypothesis that these processes derive from a common ancestral mechanism that controls expression of invading nucleic acid molecules at the post-transcriptional level. As opposed to rde-1 and qde-2 mutants, which are viable, ago1 mutants display several developmental abnormalities, including sterility. These results raise the possibility that PTGS, or at least some of its elements, could participate in the regulation of gene expression during development in plants. PMID- 11016955 TI - The endocytic receptor protein LRP also mediates neuronal calcium signaling via N methyl-D-aspartate receptors. AB - The low density lipoprotein receptor-related protein (LRP) is an endocytic receptor that is a member of the low density lipoprotein receptor family. We report that the LRP ligand, activated alpha(2)-macroglobulin (alpha(2)M*), induces robust calcium influx in cultured primary neurons, but not in nonneuronal LRP-containing cells in the same culture. The calcium influx is mediated through N-methyl-d-aspartate receptor channels, which explains the neuron specificity of the response. Microapplication of alpha(2)M* leads to a localized response at the site of application that dissipates rapidly, suggesting that the calcium signal is temporally and spatially discrete. Calcium influx to alpha(2)M* is blocked by the physiological LRP inhibitor, receptor-associated protein. Bivalent antibodies to the extracellular domain of LRP, but not Fab fragments of the same antibody, cause calcium influx, indicating that the response is specific to LRP and may require dimerization of the receptor. Thus, LRP is an endocytic receptor with a novel signaling role. PMID- 11016957 TI - Lineage-specific loss and divergence of functionally linked genes in eukaryotes. AB - By comparing 4,344 protein sequences from fission yeast Schizosaccharomyces pombe with all available eukaryotic sequences, we identified those genes that are conserved in S. pombe and nonfungal eukaryotes but are missing or highly diverged in the baker's yeast Saccharomyces cerevisiae. Since the radiation from the common ancestor with S. pombe, S. cerevisiae appears to have lost about 300 genes, and about 300 more genes have diverged by far beyond expectation. The most notable feature of the set of genes lost in S. cerevisiae is the coelimination of functionally connected groups of proteins, such as the signalosome and the spliceosome components. We predict similar coelimination of the components of the posttranscriptional gene-silencing system that includes the recently identified RNA-dependent RNA polymerase. Because one of the functions of posttranscriptional silencing appears to be "taming" of retrotransposons, the loss of this system in yeast could have triggered massive retrotransposition, resulting in elimination of introns and subsequent loss of spliceosome components that become dispensable. As the genome database grows, systematic analysis of coordinated gene loss may become a general approach for predicting new components of functional systems or even defining previously unknown functional complexes. PMID- 11016956 TI - Misregulation of gene expression in primary fibroblasts lacking poly(ADP-ribose) polymerase. AB - Poly(ADP-ribose) polymerase (PARP) is implicated in the maintenance of genomic integrity, given that inhibition or depletion of this enzyme increases genomic instability in cells exposed to genotoxic agents. We previously showed that immortalized fibroblasts derived from PARP(-/-) mice exhibit an unstable tetraploid population, and partial chromosomal gains and losses in PARP(-/-) mice and immortalized fibroblasts are accompanied by changes in the expression of p53, Rb, and c-Jun, as well as other proteins. A tetraploid population has also now been detected in primary fibroblasts derived from PARP(-/-) mice. Oligonucleotide microarray analysis was applied to characterize more comprehensively the differences in gene expression between asynchronously dividing primary fibroblasts derived from PARP(-/-) mice and their wild-type littermates. Of the 11,000 genes monitored, 91 differentially expressed genes were identified. The loss of PARP results in down-regulation of the expression of several genes involved in regulation of cell cycle progression or mitosis, DNA replication, or chromosomal processing or assembly. PARP deficiency also up-regulates genes that encode extracellular matrix or cytoskeletal proteins that are implicated in cancer initiation or progression or in normal or premature aging. These results provide insight into the mechanism by which PARP deficiency impairs mitotic function, thereby resulting in the genomic alterations and chromosomal abnormalities as well as in altered expression of genes that may contribute to genomic instability, cancer, and aging. PMID- 11016958 TI - Presynaptic N-methyl-D-aspartate receptors at the parallel fiber-Purkinje cell synapse. AB - At the cerebellar synapse between the parallel fibers (PFs) and the Purkinje cells in the cerebellum, we have found that application of N-methyl-d-aspartate (NMDA) reversibly depresses the postsynaptic current. We present evidence that this depression involves NMDA receptors located on the presynaptic axons and requires that the NMDA application be combined with action potentials in the PFs. Unexpectedly, unlike other modulations mediated by presynaptic receptors, the NMDA-induced inhibition does not involve a depression of transmitter release. Because it is blocked by both nitric oxide synthase and soluble guanylate cyclase inhibitors, we propose that it involves a trans-synaptic mechanism in which NO released by the PFs decreases the glutamate sensitivity of the Purkinje cell. PMID- 11016959 TI - Cloning of a type I cytokine receptor most related to the IL-2 receptor beta chain. AB - We have identified a type I cytokine receptor, which we have termed novel interleukin receptor (NILR), that is most related to the IL-2 receptor beta chain (IL-2Rbeta) and physically adjacent to the IL-4 receptor alpha chain gene on chromosome 16. NILR mRNA is most highly expressed in thymus and spleen, and is induced by phytohemagglutinin in human peripheral blood mononuclear cells. NILR protein was detected on human T cell lymphotropic virus type I-transformed T cell lines, Raji B cells, and YT natural killer-like cells. Artificial homodimerization of the NILR cytoplasmic domain confers proliferation to Ba/F3 murine pro-B cells but not to 32D myeloid progenitor cells or CTLL-2 murine helper T cells. In these latter cells, heterodimerization of IL-2Rbeta and the common cytokine receptor gamma chain (gamma(c)) cytoplasmic domains allows potent proliferation, whereas such heterodimerization of NILR with gamma(c) does not. This finding suggests that NILR has signaling potential but that a full understanding of its signaling partner(s) is not yet clear. Like IL-2Rbeta, NILR associates with Jak1 and mediates Stat5 activation. PMID- 11016960 TI - Plasmid-encoded MucB protein is a DNA polymerase (pol RI) specialized for lesion bypass in the presence of MucA', RecA, and SSB. AB - Replication through damaged sites in DNA requires in Escherichia coli the SOS stress-inducible DNA polymerase V (UmuC), which is specialized for lesion bypass. Homologs of the umuC gene were found on native conjugative plasmids, which often carry multiple antibiotic-resistant genes. MucB is a UmuC homolog present on plasmid R46, and its variant plasmid pKM101 has been introduced into Salmonella strains for use in the Ames test for mutagens. Using a translesion replication assay based on a gapped plasmid carrying a site-specific synthetic abasic site in the single-stranded DNA region, we show that MucB is a DNA polymerase, termed pol RI, which is specialized for lesion bypass. The activity of pol RI requires the plasmid-encoded MucA' protein and the E. coli RecA and single-strand DNA binding proteins. Elimination of any of the proteins from the reaction abolished lesion bypass and polymerase activity. The unprocessed MucA could not substitute for MucA' in the bypass reaction. The presence of a lesion bypass DNA polymerase on a native conjugative plasmid, which has a broad host range specificity and carries multiple antibiotic-resistant genes, raises the possibility that mutagenesis caused by pol RI plays a role in the spreading of antibiotic resistance among bacterial pathogens. PMID- 11016961 TI - On the tryptophan residue of smooth muscle myosin that responds to binding of nucleotide. AB - Initially, we asked which (of 10) smooth muscle myosin head residues responds to MgADP or MgATP binding with enhanced fluorescence emission (Trp-441 and Trp-512 were leading candidates)? To decide, we prepared sham-mutated smooth muscle heavy meromyosin (HMM), W441F HMM, and W512F HMM. On adding MgATP, emission of wild type and W441F HMMs increased by 25-27%, but that of W512F HMM by 5%. So, in myosin, 512 is the "sensitive Trp." Unexpectedly, properties of W512F HMM [elevated Ca(2+)-ATPase, depressed EDTA (K(+))-ATPase, no regulation of its basal or actin-activated Mg(2+)-ATPase by phosphorylation of its "regulatory" light chain, limited actin activation, and inability to move actin filaments in a motility assay] are strikingly like those of smooth muscle myosin reacted at Cys 717 with thiol reagent. From crystallography-based [Houdusse, A., Kalabakis, V. N., Himmel, D., Szent-Gyorgyi, A. G. & Cohen, C. (1999) Cell 97, 459-470] simulations, we found that in wild-type HMM with MgADP added, Trp-512 is in a "hydrophobic pocket," but that pocket becomes distorted in W512F HMM. We think that there is a "path of influence" from 512 to 717 to the active site. We suggest that the mutational changes at 512 are transmitted along this path to Cys 717, where they induce changes similar to those caused by reacting wild-type HMM with thiol reagent. PMID- 11016962 TI - IL-2-induced activation-induced cell death is inhibited in IL-15 transgenic mice. AB - A transgenic (Tg) mouse expressing human IL-15 was generated to define the role of IL-15 in the normal immune response. Overexpression of IL-15 resulted in an increase of NK, CD44(hi)CD8 memory T cells, and gammadelta T cells. Additionally, we observed the emergence of a novel type of NK-T cells with CD8alphaalpha' expression. Due to the expansion and activation of NK cells, the IL-15Tg mouse showed enhanced innate immunity. In adaptive T cell immunity, the roles of IL-15 contrasted with those of IL-2. IL-15 inhibited IL-2-induced T cell death, which plays a role in the maintenance of peripheral self-tolerance. IL-15 thus seems to contribute to enhanced immune memory by selectively propagating memory T cells and by blocking T cell death mediated by IL-2. PMID- 11016963 TI - L-serine and glycine serve as major astroglia-derived trophic factors for cerebellar Purkinje neurons. AB - Glial cells support the survival and development of central neurons through the supply of trophic factors. Here we demonstrate that l-serine (l-Ser) and glycine (Gly) also are glia-derived trophic factors. These amino acids are released by astroglial cells and promote the survival, dendritogenesis, and electrophysiological development of cultured cerebellar Purkinje neurons. Although l-Ser and Gly are generally classified as nonessential amino acids, 3 phosphoglycerate dehydrogenase (3PGDH), a key enzyme for their biosynthesis, is not expressed in Purkinje neurons. By contrast, the Bergman glia, a native astroglia in the cerebellar cortex, highly expresses 3PGDH. These data suggest that l-Ser and Gly mediate the trophic actions of glial cells on Purkinje neurons. PMID- 11016964 TI - Deformation and seismicity of Taiwan. AB - 14C-dated Holocene coastal uplift, conventional and satellite geodetic measurements, and coseismic and aseismic fault slip reveal the pattern of distributed deformation at Taiwan resulting from convergence between the Philippine Sea plate and Eurasia; as in other subduction orogenic settings, the locus of strain release and accumulation is strongly influenced by changes in fault geometry across strike. Uplift evidence from the islands of Lutao and Lanhsu is consistent with progressive oblique collision between the Luzon arc and the Chinese continental margin. In the Coastal Range, geodetic and seismic records show that shortening is taken up serially by discontinuous slip on imbricate faults. The geodetic data point to net extension across the Central Range, but deformed Holocene shorelines in the Hengchun Peninsula at its southern extremity suggest that the extension is a superficial effect partly caused by blind reverse faulting. The fastest shortening rates indicated by geodesy are recorded on the Longitudinal Valley fault and across the Chukou fault within the fold-and-thrust belt. In the former, the strain is dissipated mainly as aseismic reverse and strike-slip displacement. In contrast, the fold-and-thrust belt has witnessed five earthquakes with magnitudes of 6.5 or above in the 20th century, including the 1999.9.21 Chi-Chi earthquake (magnitude approximately 7.6) on a branch of the Chukou fault. The neotectonic and geodetic data for Taiwan as a whole suggest that the fold-and-thrust belt will continue to host the majority of great earthquakes on the island. PMID- 11016965 TI - Adaptive regulation of intestinal lysine metabolism. AB - The metabolism of dietary essential amino acids by the gut has a direct effect on their systemic availability and potentially limits growth. We demonstrate that, in neonatal pigs bearing portal and arterial catheters and fed a diet containing 23% protein [high protein (HP) diet], more than half the intake of essential amino acids is metabolized by the portal-drained viscera (PDV). Intraduodenal or i.v. infusions of [U-(13)C]-lysine were used to measure the appearance across and the use of the tracer by the PDV. In HP-fed pigs, lysine use by the PDV was derived almost entirely from the arterial input. In these animals, the small amount of dietary lysine used in first pass was oxidized almost entirely. Even so, intestinal lysine oxidation (24 micromol/kg per h) accounted for one-third of whole-body lysine oxidation (77 micromol/kg per h). Total lysine use by the PDV was not affected by low protein (LP) feeding (HP, 213 micromol/kg per h; LP,186 micromol/kg per h). In LP-fed pigs, the use of lysine by the PDV accounted for more than 75% of its intake. In contrast to HP feeding, both dietary and arterial lysines were used by the PDV of LP-fed pigs in nearly equal amounts. Intestinal lysine oxidation was suppressed completely. We conclude that the PDV are key organs with respect to amino acid metabolism and that the intestines use a disproportionately large amount of the dietary supply of amino acids during protein restriction. PMID- 11016966 TI - Three conformational states of scallop myosin S1. AB - We have determined the structure of the intact scallop myosin head, containing both the motor domain and the lever arm, in the nucleotide-free state and in the presence of MgADP.V04, corresponding to the transition state. These two new structures, together with the previously determined structure of scallop S1 complexed with MgADP (which we interpret as a detached ATP state), reveal three conformations of an intact S1 obtained from a single isoform. These studies, together with new crystallization results, show how the conformation of the motor depends on the nucleotide content of the active site. The resolution of the two new structures ( approximately 4 A) is sufficient to establish the relative positions of the subdomains and the overall conformation of the joints within the motor domain as well as the position of the lever arm. Comparison of available crystal structures from different myosin isoforms and truncated constructs in either the nucleotide-free or transition states indicates that the major features within the motor domain are relatively invariant in both these states. In contrast, the position of the lever arm varies significantly between different isoforms. These results indicate that the heavy-chain helix is pliant at the junction between the converter and the lever arm and that factors other than the precise position of the converter can influence the position of the lever arm. It is possible that this pliant junction in the myosin head contributes to the compliance known to be present in the crossbridge. PMID- 11016967 TI - Evidence from sequence-tagged-site markers of a recent progenitor-derivative species pair in conifers. AB - Black spruce (Picea mariana [B.S.P.] Mill.) and red spruce (Picea rubens Sarg.) are two conifer species known to hybridize naturally in northeastern North America. We hypothesized that there is a progenitor-derivative relationship between these two taxa and conducted a genetic investigation by using sequence tagged-site markers of expressed genes. Based on the 26 sequence-tagged-site loci assayed in this study, the unbiased genetic identity between the two taxa was quite high with a value of 0.920. The mean number of polymorphic loci, the mean number of alleles per polymorphic locus, and the average observed heterozygosity were lower in red spruce (P = 35%, A(P) = 2.1, H(o) = 0.069) than in black spruce (P = 54%, A(P) = 2.9, H(o) = 0.103). No unique alleles were found in red spruce, and the observed patterns of allele distribution indicated that the genetic diversity of red spruce was essentially a subset of that found in black spruce. When considered in combination with ecological evidence and simulation results, these observations clearly support the existence of a progenitor-derivative relationship and suggest that the reduced level of genetic diversity in red spruce may result from allopatric speciation through glaciation-induced isolation of a preexisting black spruce population during the Pleistocene era. Our observations signal a need for a thorough reexamination of several conifer species complexes in which natural hybridization is known to occur. PMID- 11016968 TI - Generation of oscillations by the p53-Mdm2 feedback loop: a theoretical and experimental study. AB - The intracellular activity of the p53 tumor suppressor protein is regulated through a feedback loop involving its transcriptional target, mdm2. We present a simple mathematical model suggesting that, under certain circumstances, oscillations in p53 and Mdm2 protein levels can emerge in response to a stress signal. A delay in p53-dependent induction of Mdm2 is predicted to be required, albeit not sufficient, for this oscillatory behavior. In line with the predictions of the model, oscillations of both p53 and Mdm2 indeed occur on exposure of various cell types to ionizing radiation. Such oscillations may allow cells to repair their DNA without risking the irreversible consequences of continuous excessive p53 activation. PMID- 11016969 TI - Involvement of neurogranin in the modulation of calcium/calmodulin-dependent protein kinase II, synaptic plasticity, and spatial learning: a study with knockout mice. AB - Neurogranin/RC3 is a neural-specific Ca(2+)-sensitive calmodulin (CaM)-binding protein whose CaM-binding affinity is modulated by phosphorylation and oxidation. Here we show that deletion of the Ng gene in mice did not result in obvious developmental or neuroanatomical abnormalities but caused an impairment of spatial learning and changes in hippocampal short- and long-term plasticity (paired-pulse depression, synaptic fatigue, long-term potentiation induction). These deficits were accompanied by a decreased basal level of the activated Ca(2+)/CaM-dependent kinase II (CaMKII) ( approximately 60% of wild type). Furthermore, hippocampal slices of the mutant mice displayed a reduced ability to generate activated CaMKII after stimulation of protein phosphorylation and oxidation by treatments with okadaic acid and sodium nitroprusside, respectively. These results indicate a central role of Ng in the regulation of CaMKII activity with decisive influences on synaptic plasticity and spatial learning. PMID- 11016970 TI - Targeted disruption of the galanin gene reduces the number of sensory neurons and their regenerative capacity. AB - The neuropeptide galanin is expressed developmentally in the dorsal root ganglion (DRG) and is rapidly up-regulated 120-fold after peripheral nerve section in the adult. Here we report that adult mice carrying a loss-of-function mutation in the galanin gene have a 13% reduction in the number of cells in the DRG associated with a 24% decrease in the percentage of neurons that express substance P. These deficits are associated with a 2.8- and 2.6-fold increase in the number of apoptotic cells in the DRG at postnatal days 3 and 4, respectively. After crush injury to the sciatic nerve, the rate of peripheral nerve regeneration is reduced by 35% with associated long-term functional deficits. Cultured DRG neurons from adult mutant mice demonstrate similar deficits in neurite number and length. These results identify a critical role for galanin in the development and regeneration of sensory neurons. PMID- 11016972 TI - The case of medium-dependent dual mechanisms for photoisomerization: one-bond flip and hula-twist. AB - This paper critically reviews examples in the literature of photochemical cis trans isomerization paying particular attention to the medium effect and accompanied conformational changes. A case is made that the Hula-Twist mechanism, postulated in 1985 as a photochemical reaction pathway for a polyene chromophore imbedded in a protein binding cavity such as those of rhodopsin and bacteriorhodopsin, is also a dominant reaction pathway for a diene, or a longer polyene confined in a rigid (relative to isomerization rate) medium. The conventional one-bond-flip process is the preferred reaction pathway in a fluid medium. While defining experiments are proposed, this dual mechanistic approach successfully accounts for all examples in the literature on photoisomerization reactions whether involving conformational changes or not. PMID- 11016971 TI - Galanin regulates the postnatal survival of a subset of basal forebrain cholinergic neurons. AB - The neuropeptide galanin colocalizes with choline acetyltransferase, the synthetic enzyme for acetylcholine, in a subset of cholinergic neurons in the basal forebrain of rodents. Chronic intracerebroventricular infusion of nerve growth factor induces a 3- to 4-fold increase in galanin gene expression in these neurons. Here we report the loss of a third of cholinergic neurons in the medial septum and vertical limb diagonal band of the basal forebrain of adult mice carrying a targeted loss-of-function mutation in the galanin gene. These deficits are associated with a 2-fold increase in the number of apoptotic cells in the forebrain at postnatal day seven. This loss is associated with marked age dependent deficits in stimulated acetylcholine release, performance in the Morris water maze, and induction of long-term potentiation in the CA1 region of the hippocampus. These data provide unexpected evidence that galanin plays a trophic role to regulate the development and function of a subset of septohippocampal cholinergic neurons. PMID- 11016973 TI - Histone H2A-mediated transient cytokine gene delivery induces efficient antitumor responses in murine neuroblastoma. AB - A major goal of cancer immunotherapy is the induction of a cell-mediated antitumor response in poorly immunogenic malignancies. We tested the hypothesis that this can be achieved by cytokine gene therapy with a novel histone H2A-based transient transfection procedure. This was tested by using cytokine genes encoding for IL-2 and a single chain IL-12 (scIL-12) fusion protein in a recently developed murine neuroblastoma model. Here, we demonstrate that cytokine gene transfer of IL-2 and scIL-12 with histone H2A results in the induction of an antitumor immune response that is superior in some respects to gene transfer with Superfect, a commercially available activated dendrimer commonly used to effect transfection with plasmids. Three lines of evidence support this contention. First, histone H2A-mediated transfection of IL-2 induces a natural killer cell induced rejection of primary tumors in contrast to Superfect, which produces only a partial reduction in primary tumor growth. Second, the induction of a T cell mediated protective tumor immunity following gene transfer of scIL-12 is more efficient with the histone H2A-mediated gene transfer because rejection of a lethal wild-type tumor cell challenge is accompanied by the greatest degree of MHC class I-restricted tumor cell killing in vitro. Third, histone H2A-mediated scIL-12 gene therapy induces the greatest release of mIFN-gamma from splenocytes of vaccinated animals in contrast to Superfect and other controls. PMID- 11016974 TI - DNA damage, DNA repair, cell proliferation, and DNA replication: how do gene mutations result? PMID- 11016975 TI - Hindering auxiliary anchors are potent modulators of peptide binding and selection by I-Ak class II molecules. AB - Selection of particular antigen-derived peptides by class II MHC molecules determines the population of complexes represented on the antigen-presenting cell surface and available for T cell receptor engagement. This discriminating selection process results from unique interactions between the spectrum of peptides generated during antigen processing and the MHC molecules. Here, we examined the selection of peptides by the class II MHC, I-A(k). Our results indicate that although peptide primary anchors are key in MHC binding, auxiliary anchors are a powerful regulatory component in the selection of peptides by I A(k). Study of the segments surrounding the dominant hen egg white lysozome(48 61) epitope demonstrates that auxiliary anchors also are involved in determining the binding register of I-A(k) along an extended peptide. In addition, we found that unique combinations of auxiliary anchors can act in concert to modulate the binding of peptides to I-A(k). PMID- 11016976 TI - Mobile elements and chromosomal evolution in the virilis group of Drosophila. AB - Species of the virilis group of Drosophila differ by multiple inversions and chromosome fusions that probably accompanied, or led to, speciation. Drosophila virilis has the primitive karyotype for the group, and natural populations are exceptional in having no chromosomal polymorphisms. We report that the genomic locations of Penelope and Ulysses transposons are nonrandomly distributed in 12 strains of D. virilis. Furthermore, Penelope and Ulysses insertion sites in D. virilis show a statistically significant association with the breakpoints of inversions found in other species of the virilis group. Sixteen newly induced chromosomal rearrangements were isolated from the progeny of D. virilis hybrid dysgenic crosses, including 12 inversions, 2 translocations, and 2 deletions. Penelope and Ulysses were associated with the breakpoints of over half of these new rearrangements. Many rearrangement breakpoints also coincide with the chromosomal locations of Penelope and Ulysses insertions in the parental strains and with breakpoints of inversions previously established for other species of the group. Analysis of homologous sequences from D. virilis and Drosophila lummei indicated that Penelope insertion sites were closely, but not identically, located at the nucleotide sequence level. Overall, these results indicate that Penelope and Ulysses insert in a limited number of genomic locations and are consistent with the possibility that these elements play an important role in the evolution of the virilis species group. PMID- 11016977 TI - Telomere fusions caused by mutating the terminal region of telomeric DNA. AB - Mutations in the template region of a telomerase RNA gene can lead to the corresponding sequence alterations appearing in newly synthesized telomeric repeats. We analyzed a set of mutations in the template region of the telomerase RNA gene (TER1) of the budding yeast Kluyveromyces lactis that were predicted to lead to synthesis of mutant telomeric repeats with disrupted binding of the telomeric protein Rap1p. We showed previously that mutating the left side of the 12-bp consensus Rap1p binding site led to immediate and severe telomere elongation. Here, we show that, in contrast, mutating either the right side of the site or both sides together leads initially to telomere shortening. On additional passaging, certain mutants of both categories exhibit telomere telomere fusions. Often, six new Bal-31-resistant, telomere repeat-containing bands appeared, and we infer that each of the six K. lactis chromosomes became circularized. These fusions were not stable, appearing occasionally to resolve and then reform. We demonstrate directly that a linear minichromosome introduced into one of the fusion mutant strains circularized by means of end-to-end fusions of the mutant repeat tracts. In contrast to the chromosomal circularization reported previously in Schizosaccharomyces pombe mutants defective in telomere maintenance, the K. lactis telomere fusions retained their telomeric DNA repeat sequences. PMID- 11016978 TI - Myxococcus xanthus fibril appendages are essential for excitation by a phospholipid attractant. AB - Isolated (A-motile) Myxococcus xanthus cells glide over solid surfaces and display excitation, a suppression of direction reversals, when presented with phosphatidylethanolamine (PE) purified from its own membranes or synthetic dilauroyl PE and dioleoyl PE. Although the mechanism of PE signal transduction is unknown, we hypothesized that M. xanthus might use surface-associated factors to detect exogenous PE to prevent endogenous lipids from self-stimulating the sensory system. Peritrichous protein and polysaccharide appendages called fibrils were correlated with dilauroyl PE excitation. Wild-type cells treated with Congo red, an inhibitor of fibril assembly, and mutants defective in fibril biosynthesis showed an elevated reversal period, which suggested that fibrils regulate the gliding motor. Furthermore, the loss of fibrils resulted in loss of excitation to dilauroyl PE but not dioleoyl PE. Restoration of fibril production to these mutants restored the dilauroyl PE response. In addition, the dif cytoplasmic signal transduction system and starvation conditions were required for dilauroyl PE excitation. The chemically specific nature of the response and the dependence on the dif system suggests that fibrils define a novel sensory organelle whose evolution may have been necessary to prevent autostimulation by endogenous membrane lipids. Because the hydrophobic nature of dilauroyl PE would be inaccessible to periplasmic chemosensors, we suggest that fibrils act as extracellular signal transducers to probe surfaces for insoluble chemical signals. PMID- 11016979 TI - Cellular mechanisms of myogenic activity in gastric smooth muscle. AB - In many regions of the intestine, a thin layer of interstitial cells of Cajal (ICC) lie in the myenteric region, between the circular and longitudinal muscle layers. ICC are connected by gap junctions to surrounding ICC and also with circular and longitudinal smooth muscle cells, forming a large electrical syncytium. Damage of the ICC causes a disorder in the patterns of rhythmic activity. Isolated ICC produce a rhythmic oscillation of the membrane potential. All these observations have led to the suggestion that ICC may be the pacemaker cell responsible for intestinal activity. Gastric smooth muscles generate slow oscillatory membrane potential changes (slow waves) and spike potentials. The activity is considered to be linked to the metabolism in the cell. Three types of cells located in the gastric wall (circular and longitudinal smooth muscle cells and ICC) produce synchronized electrical responses with different shapes. The electrical responses appear to originate in ICC and then spread to the smooth muscle layers, indicating that ICC may also be the pacemaker cells responsible for gastric activity. However, isolated circular smooth muscle tissues spontaneously generate regenerative potentials, suggesting that there are at least two sites for the initiation of spontaneous activity in the stomach. Regenerative potentials persist in the presence of Ca-antagonists and are inhibited by agents which disrupt intracellular Ca(2+) homeostasis. Depolarization of the membrane elicits regenerative potentials after a long delay and the potentials have long refractory periods. This suggests that an unidentified 2nd messenger may be formed during the delay between membrane depolarization and the initiation of a regenerative potential. In gastric muscles of mutant mice which do not express inositol trisphosphate (InsP(3)) receptors, spike potentials but not slow waves are generated, suggesting the possible involvement of InsP(3) in the initiation of spontaneous activity. PMID- 11016980 TI - Neuromuscular adaptation to microgravity environment. AB - Morphological and/or functional char-acteristics of skeletal muscles have a greater adaptability in response to changes in environmental stimuli. For example, an atrophy associated with a shift of fiber characteristics toward fast twitch type is a common adaptation of antigravity muscle to a microgravity environment. Neuromuscular responses and possible mechanisms of both neural and muscular adaptations to a microgravity environment are discussed in this article. Responses of morphological, metabolic, and contractile properties, as well as fiber phenotype, of muscles are briefly reviewed. Discussion is further extended to the patterns of electromyogram and tension development of muscle, responses of postural stability and locomotion, and/or motoneurons in order to study the mechanism for muscular adaptation to microgravity. PMID- 11016981 TI - Body sway induced by depth linear vection in reference to central and peripheral visual field. AB - PURPOSE: A significant correlation between the magnitude of linear vection and the degree of body sway induced by a visual stimulus perceived as moving in depth was previously described (Jpn J Physiol 49: 417-424, 1999). The purpose of this study was to examine the role of the central and peripheral visual fields in inducing vection and body sway. METHODS: Ten healthy volunteer students who had no vestibular or neurological disorders served as subjects. A depth optokinetic stimulus (DOKS) was projected onto a head-mounted display (HMD) and was perceived to move in depth. Different amounts of the central or peripheral visual field were masked independently. The magnitude of the linear vection induced by the DOKS was evaluated by verbal assessment and compared with the magnitude of induced body sway. Body sway was monitored by a video-motion-analyzer that recorded the movement of the head, shoulder, hip, knee and ankle. RESULTS: The magnitude of vection was correlated with the frequency of DOKS and also with the amplitude of body sway (r = 0.74). When the central visual field was restricted by 10 to 30%, there was almost no change in the induced body sway and vection. However, when central occlusion was greater than 40%, depth perception and induced body movement were greatly reduced. With increasing amounts of peripheral field occlusion from 50 to 90%, there was a greater reduction of both vection and body sway. CONCLUSION: Vection is strongly correlated with body movement, and vection and body sway were more dependent on stimulation of the peripheral visual field. PMID- 11016982 TI - Stimulation of Cl(-) secretion by L-alanine oligopeptides in the mammalian large intestine. AB - A variety of oligopeptides are probably released within the intestinal tissue under inflammatory conditions or during peptide absorption. To examine whether some of these peptides can affect intestinal transport functions, we determined the effects of L-alanine oligopeptide on short-circuit current (I(sc)) and transmucosal conductance (G(t)) in submucosa-mucosa preparations from the mouse cecum and guinea pig distal colon in vitro in Ussing chambers. L-Alanyl-L-alanine (Ala-Ala, 10 mM) added to the serosal side increased I(sc) and G(t), giving a peak followed by a sustained phase (the peak increase in I(sc) was 45 +/-6 microA/cm(2) and the increase in G(t) was 0.55+/-0.11 mS/cm(2)). The tripeptide, L-alanyl-L-alanyl-L-alanine (Ala-Ala-Ala, 10 mM), added to the serosal side also induced increases in I(sc) and G(t) by a similar degree. On the other hand, luminal Ala-Ala, and serosal L-alanine and L-alanine (10 mM) caused significantly smaller increases in I(sc) and G(t) ( approximately 15 microA/cm(2) and approximately 0. 15 mS/cm(2), respectively). The Ala-Ala induced increase in I(sc) was partially inhibited by serosal bumetanide (0.1 mM) and mucosal 5-nitro 2-(3-phenylpropylamino)benzoic acid (0.1 mM), and largely suppressed by removing Cl(-) from the bathing solution. The increase in I(sc) was largely suppressed by serosal low Ca(2+) and tetrodotoxin, but was not affected by indomethacin. In the guinea pig distal colon, serosal Ala-Ala (10 mM) evoked a transient increase in I(sc) by 23+/-7 microA/cm(2) and an increase in G(t) by 1.2+/-0.3 mS/cm(2). These results suggest that Ala-Ala, and probably also Ala-Ala-Ala, added to the serosal side stimulated electrogenic Cl(-) secretion mainly through the activation of submucosal secretomotor neurons in the mammalian large intestine. PMID- 11016983 TI - High pressure sensitizes murine erythroleukemia cells to caffeine-induced premature mitosis. AB - Murine erythroleukemia (MEL) cells were exposed to a high pressure of 80 MPa or aphidicolin (APH), DNA polymerase inhibitor. The effects of caffeine on cell cycle were examined using these cells. During the culture of 80 MPa-treated MEL cells at atmospheric pressure, the cells arrested in the G2 phase, and cyclin B and hyperphosphorylated p34(cdc2) were accumulated. Namely, maturation promoting factor (MPF) composed of p34(cdc2) and cyclin B was inactive. However, upon exposure to caffeine, these cells entered prematurely into mitosis by activating MPF. Caffeine-induced premature mitosis was suppressed by butyrolactone I and orthovanadate. On the other hand, APH-treated MEL cells, which were not exposed to 80 MPa, were not so sensitive to caffeine-induced premature mitosis despite cyclin B accumulation. In this case, dephosphorylation of p34(cdc2) was not induced by caffeine. Interestingly, caffeine-induced premature mitosis in the 80 MPa-treated cells was also suppressed by APH. These results suggest that the premature mitosis of 80 MPa-treated MEL cells by caffeine is induced by active MPF, and that APH-sensitive molecules such as DNA polymerase may also play an important role in the checkpoint that controls the transition from G2 to M phase. PMID- 11016984 TI - Nitric oxide and thermogenic function of brown adipose tissue in rats. AB - To clarify the effects of cold acclimation and immobilization stress adaptation of rats on nitric oxide (NO) activity in interscapular brown adipose tissue (BAT), we incubated neatly diced (1-mm(3) blocks) BAT in a metabolic chamber for respiration, measured oxygen consumption using a Clark electrode, and estimated NO release in the buffer medium by measuring nitrite plus nitrate (NO(x)) using the Griess method (diazotization reaction). The production of NO(x) in the buffer medium confirmed that BAT releases NO, as there is no other source of NO(x) in the system. The NO activity was observed in the basal condition and increased with noradrenaline stimulation, showing a correlation with oxygen consumption in the warm (25 degrees C)-acclimated control rats. Cold acclimation (5 degrees C, 5 weeks) or immobilization stress adaptation (3 h daily, 25 degrees C, 5 weeks) caused enhanced NO activity in the basal condition in comparison with the control. We suggest that NO is involved in enhancement of the thermogenic functions of BAT in rats. PMID- 11016986 TI - Purkinje cell activity in the middle zone of the cerebellar flocculus during optokinetic and vestibular eye movement in cats. AB - Based on the inverse dynamics theory, a previous paper reconstructed simple-spike (SS) firing rates of Purkinje cells in the cat's flocculus middle-zone by a linear-weighted summation of eye acceleration, velocity, and position during optokinetic response (OKR). The present study investigated the SS rates during combined optokinetic and vestibular stimuli of the cells recorded in the previous paper. During the sinusoidal vestibuloocular reflex (VOR) in the light (VORL) and in the dark (VORD) the firing modulation was small. During VOR suppression (VORS) by head and visual-pattern rotation in the same direction, the modulation was deep, with the peak coinciding roughly with peak ipsiversive head velocity. During VOR enhancement (VORE), the modulation was deep, with the peak coinciding roughly with peak contraversive head velocity. If we interpret these data in relation to eye and head movements, the cells in the cat were comparable to the horizontal-gaze-velocity Purkinje cells in the monkey that encode a linear summation of eye and head velocity signals. Alternatively, if we interpret the data on the basis of the inverse dynamics theory, the SS rates during VORL, VORS, and VORE were well-fitted by the OKR components of the movements (subtraction of VORD from VORL, VORS, and VORE eye movements, respectively), but not by the whole movements, using the coefficients calculated during OKR. It is concluded that the data are interpretable by both theories when the VOR gain (eye movement/head movement) is close to 1 and the firing is dominated by eye velocity information. PMID- 11016985 TI - Effects of VEGF on Ca(2+)-transient in cultured lymphatic endothelial cells and mechanical activity of isolated lymph vessels. AB - We investigated the effects of vascular endothelial growth factor (VEGF(165)) on [Ca(2+)](i)-transient in cultured lymphatic endothelial cells (LEC) and mechanical activity of isolated dog thoracic ducts. VEGF (0.1-10 ng/ml) caused a dose-dependent increase of the [Ca(2+)](i) in LEC. Pretreatment with 10(-5) M genistein or 5x10(-6) M herbimycin A produced a significant reduction of the VEGF induced [Ca(2+)](i)-transient. In the presence of 10(-6) M thapsigargin, VEGF caused no significant effect on the [Ca(2+)](i)-transient. Pretreatment with Ca(2+)-free solution containing 0.1 mM EGTA produced no significant effect on the peak increase of [Ca(2+)](i) induced by 0.1 or 10 ng/ml VEGF, but significantly depressed the sustained part of [Ca(2+)](i) observed at the higher concentration of VEGF. The VEGF (0.1-10 ng/ml) caused a significant dilation of the isolated lymph vessels with intact endothelium, which were precontracted with U46,619. The 10 ng/ml VEGF-induced dilation was significantly reduced by 3 x 10(-5) M N(omega) nitro-L-arginine methyl ester (L-NAME). The action of L-NAME was inhibited by the simultaneous application of 10(-3) M L-arginine. Mechanical rubbing of the endothelium also caused significant inhibition of the VEGF-induced dilation. The findings suggest that VEGF(165) may activate the receptor-related tyrosine kinase and cause the release of Ca(2+) from the inositol 1,4, 5-triphosphate-sensitive intracellular Ca(2+) stores in LEC. VEGF(165) also produces endothelium-dependent nitric oxide-mediated dilation of the precontracted isolated lymph vessels. PMID- 11016987 TI - Closed-loop identification of carotid sinus baroreflex transfer characteristics using electrical stimulation. AB - Although random aortic pressure (AOP) perturbation according to a binary white noise sequence enables us to estimate open-loop dynamic characteristics of the carotid sinus baroreflex under closed-loop conditions, the necessity of arterial catheter implantation limits the applicability of this method in freely moving animal experiments. Thus, we explored a closed-loop system identification method using electrical stimulation. In 6 anesthetized and vagotomized rabbits, we stimulated the aortic depressor nerve with a binary white noise sequence (0-10 Hz) under baroreflex closed-loop conditions while measuring cardiac sympathetic nerve activity (SNA) and AOP. We used a closed-loop identification method to estimate the peripheral arc transfer function from SNA to AOP. The peripheral arc transfer function approximated a second-order low-pass filter and its fitted parameters did not differ from those obtained by an open-loop identification method (dynamic gain: 1.16+/-0.32 vs. 1.02+/-0.11; natural frequency: 0.08+/-0.03 vs. 0.09+/-0.03 Hz; damping ratio: 1.53+/-0.15 vs. 1.57+/-0.21). In 6 different rabbits, we applied intermittent rapid pacing (396 beats/min) under baroreflex closed-loop conditions to estimate the neural arc transfer function from AOP to SNA. The neural arc transfer function approximated a first-order high-pass filter and its fitted parameters did not differ from those obtained by an open-loop identification method (dynamic gain: -1.15+/-0.45 vs. -1.06+/-0.05; corner frequency: 0.12+/-0.05 vs. 0.13+/-0.03 Hz). In conclusion, the closed-loop identification method using electrical stimulation is effective to estimate the neural and peripheral arc transfer functions. PMID- 11016988 TI - Metabolic and neuromuscular adaptations to endurance training in professional cyclists: a longitudinal study. AB - The aim of this longitudinal study was to analyze the changes in several metabolic and neuromuscular variables in response to endurance training during three defined periods of a full sports season (rest, precompetition and competition). The study population was formed by thirteen professional cyclists (age +/- SEM: 24+/-1 years; mean V(O2 max) approximately 74 ml kg(-1) min(-1)). In each testing session, subjects performed a ramp test until exhaustion on a cycle ergometer (workload increases of 25 W min(-1)). The following variables were recorded every 100 W until the tests: oxygen consumption (V(O2) in l min( 1)), respiratory exchange ratio (RER in V(CO2) V(O2)(-1)) and blood lactate, pH and bicarbonate concentration [HCO3(-)]. Surface electromyography (EMG) recordings were also obtained from the vastus lateralis to determine the variables: root mean square voltage (rms-EMG) and mean power frequency (MPF). RER and lactate values both showed a decrease (p<0.05) throughout the season at exercise intensities corresponding to submaximal workloads. In contrast, no significant differences were found in mean pH or [HCO(3-)]. Finally, rms-EMG tended to increase during the season, with significant differences (p<0.05) observed mainly between the competition and rest periods at most workloads. In contrast, precompetition MPF values increased (p<0.05) with respect to resting values at most submaximal workloads but fell (p<0.05) during the competition period. Our findings suggest that endurance conditioning induces the following general adaptations in elite athletes: (1) lower circulating lactate and increased reliance on aerobic metabolism at a given submaximal intensity, and possibly (2) an enhanced recruitment of motor units in active muscles, as suggested by rms-EMG data. PMID- 11016990 TI - The $30 billion punt PMID- 11016991 TI - The ecogen 66 sacrificed for the greenpeace 28 PMID- 11016989 TI - Mobilization of intracellular Ca(2+) by thromboxane A(2) does not affect Ca(2+) activated K(+) channels in rat colonic crypt cells. AB - The effects of 9,11-epithio-11,12-methano-thromboxane A(2) (STA(2)), a stable thromboxane A(2) analogue, and carbachol on colonic Ca(2+)-activated K(+) channels were studied. In indo-1-loaded single cells in isolated rat colonic crypts, both STA(2) (0.1 microM) and carbachol (10 microM) transiently increased intracellular free Ca(2+) concentration ([Ca(2+)](i)) by 136 and 155 nm, respectively. In whole-cell current-clamp experiments of the colonic crypt cells with Cl(-)-free solutions, carbachol (10 microM) hyperpolarized the cell by 19.7 mV, while STA(2) (0.1 microM) did not affect the membrane potential. In the isolated colonic mucosa that was permeabilized mucosally by a monovalent ionophore nystatin in the presence of a serosally directed K(+) gradient, carbachol (10 microM) transiently elicited K(+) current, but STA(2) (0.1 microM) did not. These results indicate that STA(2) elevates [Ca(2+)](i) in rat colonic crypt cells but does not activate basolateral Ca(2+)-activated K(+) channels. PMID- 11016992 TI - Clear vision for a structure-seeking business. PMID- 11016994 TI - Erratum PMID- 11016993 TI - Uninformation and the choice paradox. PMID- 11016996 TI - Corrigenda PMID- 11016995 TI - Laboratory testing. PMID- 11016997 TI - Objectively assessing bioartificial organs. PMID- 11016999 TI - Chiron buys PathoGenesis PMID- 11016998 TI - Bad behavior. PMID- 11017000 TI - Corning chips in PMID- 11017001 TI - Guilford halts gliatech deal PMID- 11017002 TI - GMO roundup PMID- 11017003 TI - State restricts biotech crops PMID- 11017004 TI - Research collaborations PMID- 11017005 TI - PJect buys celltech vax PMID- 11017006 TI - Biotech exec jailed PMID- 11017008 TI - Roll out the barrel scaffold PMID- 11017007 TI - IBM invests in life sciences PMID- 11017009 TI - Chromosome targeting PMID- 11017011 TI - Bt corn kills monarch? PMID- 11017010 TI - A snare for the weak PMID- 11017013 TI - A small-molecule PDGF inhibitor PMID- 11017012 TI - Rule of three PMID- 11017017 TI - Synthetic transcriptional activators PMID- 11017014 TI - Protein identification at nanomolar concentration PMID- 11017015 TI - A FACS screen for improving enzymes PMID- 11017019 TI - Bt rice makes its debut PMID- 11017018 TI - Review PMID- 11017020 TI - Mammalian artificial chromosomes PMID- 11017021 TI - Therapy for mucosal infections PMID- 11017022 TI - Electrocatalytic DNA mismatch detection PMID- 11017024 TI - A two-hybrid system for integral membrane proteins PMID- 11017023 TI - Cloned piglets parte trois PMID- 11017026 TI - Internet chat damages biotechnology stocks PMID- 11017025 TI - NIH, FDA favor better "management" of conflicts of interest PMID- 11017027 TI - Critics slam new monarch Bt-corn data PMID- 11017028 TI - Electronic postings thought to harm other stocks PMID- 11017030 TI - Italy toasts sardinia with SharDNA PMID- 11017029 TI - PERV data renew xeno debate PMID- 11017031 TI - Geron bio-Med re-directs funds away from xeno pigs PMID- 11017032 TI - UK set to allow research on embryonic stem cells. PMID- 11017034 TI - Efforts to commercialize structural genomics may be limited PMID- 11017033 TI - . . . as NIH offers guidelines for stem cell research amid broader debate. PMID- 11017035 TI - Sensing proteins outside of the box. PMID- 11017036 TI - Transgenic commensals as mucosal protectants. PMID- 11017037 TI - The single cell as a microplate well. PMID- 11017038 TI - Probing proteomes--seeing the whole picture? PMID- 11017039 TI - Electrochemical DNA analysis comes of age. PMID- 11017040 TI - Milking the genome for profit. AB - Despite recent accomplishments, significant challenges remain before cloning can become a routine part of transgenic animal production. PMID- 11017041 TI - Status of genome projects for nonpathogenic bacteria and archaea. AB - Since the first microbial genome was sequenced in 1995, 30 others have been completed and an additional 99 are known to be in progress. Although the early emphasis of microbial genomics was on human pathogens for obvious reasons, a significant number of sequencing projects have focused on nonpathogenic organisms, beginning with the release of the complete genome sequence of the archaeon Methanococcus jannaschii in 1996. The past 18 months have seen the completion of the genomes of several unusual organisms, including Thermotoga maritima, whose genome reveals extensive potential lateral transfer with archaea; Deinococcus radiodurans, the most radiation-resistant microorganism known; and Aeropyrum pernix, the first Crenarchaeota to be completely sequenced. Although the functional characterization of genomic data is still in its initial stages, it is likely that microbial genomics will have a significant impact on environmental, food, and industrial biotechnology as well as on genomic medicine. PMID- 11017042 TI - Production of cloned pigs from in vitro systems. AB - Here we describe a procedure for cloning pigs by the use of in vitro culture systems. Four healthy male piglets from two litters were born following nuclear transfer of cultured somatic cells and subsequent embryo transfer. The initiation of five additional pregnancies demonstrates the reproducibility of this procedure. Its important features include extended in vitro culture of fetal cells preceding nuclear transfer, as well as in vitro maturation and activation of oocytes and in vitro embryo culture. The cell culture and nuclear transfer techniques described here should allow the use of genetic modification procedures to produce tissues and organs from cloned pigs with reduced immunogenicity for use in xenotransplantation. PMID- 11017043 TI - Therapy of mucosal candidiasis by expression of an anti-idiotype in human commensal bacteria. AB - Two recombinant strains of Streptococcus gordonii, secreting or displaying a microbicidal single-chain antibody (H6), and stably colonizing rat vagina, were used to treat an experimental vaginitis caused by Candida albicans. A post challenge intravaginal delivery of the H6-secreting strain was as efficacious as fluconazole in rapidly abating the fungal burden. Three weeks after challenge, 75% and 37.5% of the rats treated with the H6-secreting or displaying bacteria, respectively, were cured of the infection, which persisted in 100% of the animals treated with a S. gordonii strain expressing an irrelevant single-chain antibody. Thus, a human commensal bacterium can be suitably engineered to locally release a therapeutic antibody fragment. PMID- 11017044 TI - Design of GFB-111, a platelet-derived growth factor binding molecule with antiangiogenic and anticancer activity against human tumors in mice. AB - We have designed a molecule, GFB-111, that binds to platelet-derived growth factor (PDGF), prevents it from binding to its receptor tyrosine kinase, and blocks PDGF-induced receptor autophosphorylation, activation of Erk1 and Erk2 kinases, and DNA synthesis. GFB-111 is highly potent (IC50 = 250 nM) and selective for PDGF over EGF, IGF-1, aFGF, bFGF, and HRGbeta (IC50 values > 100 microM), but inhibits VEGF-induced Flk-1 tyrosine phosphorylation and Erk1/Erk2 activation with an IC50 of 10 microM. GFB-111 treatment of nude mice bearing human tumors resulted in significant inhibition of tumor growth and angiogenesis. The results demonstrate the feasibility of designing novel growth factor-binding molecules with potent anticancer and antiangiogenic activity. PMID- 11017045 TI - Function-based isolation of novel enzymes from a large library. AB - Here we describe a high-throughput, quantitative method for the isolation of enzymes with novel substrate specificities from large libraries of protein variants. Protein variants are displayed on the surface of microorganisms and incubated with a synthetic substrate consisting of (1) a fluorescent dye (2) a positively charged moiety (3) the target scissile bond, and (4) a fluorescence resonance energy transfer (FRET) quenching partner. Enzymatic cleavage of the scissile bond results in release of the FRET quenching partner while the fluorescent product is retained on the cell surface, allowing isolation of catalytically active clones by fluorescence-activated cell sorting (FACS). Using a synthetic substrate with these characteristics, we enriched Escherichia coli expressing the serine protease OmpT from cells expressing an inactive OmpT variant by over 5,000-fold in a single round. Screening a library of 6 x 10(5) random OmpT variants by FACS using a FRET peptide substrate with a nonpreferred Arg-Val cleavage sequence resulted in the isolation of variant proteases with catalytic activities enhanced by as much as 60-fold. This approach represents a potentially widely applicable method for high-throughput screening of large libraries on the basis of catalytic turnover. PMID- 11017047 TI - Minimal activators that bind to the KIX domain of p300/CBP identified by phage display screening. AB - Human gene therapy approaches involving transcription factors often rely on artificial activation domains for transcriptional activation. These domains are often large (e.g., 80 amino acids for VP16), recruit multiple co-activation complexes at once, and offer no fine control over the level of transcription. In an attempt to understand the sequence and structural requirements of a minimal mammalian activator, we employed a molecular diversity approach with a peptide phage display library composed of random eight-amino acid peptides. Using the KIX domain of the mammalian co-activators p300 and CBP as target, we discovered a family of synthetic binding peptides. These peptides share significant homology with natural KIX domain ligands, and are shown to bind an overlapping, yet distinct, surface of p300/CREB-binding protein (CBP). When fused to a heterologous DNA binding domain, these synthetic peptides function as titratable, modular, and potent transcriptional activators in living cells through specific recruitment of p300/CBP, with the level of transcriptional activation proportional to the affinity of the synthetic peptide for the KIX domain. Taken together, our data demonstrate that a molecular diversity approach can be used to discover minimal, co-activator domain-specific synthetic activators, and that transcriptional activation can be modulated as desired at the level of co activator recruitment. PMID- 11017046 TI - Use of G-protein fusions to monitor integral membrane protein-protein interactions in yeast. AB - The control of protein-protein interactions is a fundamental aspect of cell regulation. Here we describe a new approach to detect the interaction of two proteins in vivo. By this method, one binding partner is an integral membrane protein whereas the other is soluble but fused to a G-protein gamma-subunit. If the binding partners interact, G-protein signaling is disrupted. We demonstrate interaction between known binding partners, syntaxin 1a with neuronal Sec1 (nSec1), and the fibroblast-derived growth factor receptor 3 (FGFR3) with SNT-1. In addition, we describe a genetic screen to identify nSec1 mutants that are expressed normally, but are no longer able to bind to syntaxin 1a. This provides a convenient method to study interactions of integral membrane proteins, a class of molecules that has been difficult to study by existing biochemical or genetic methods. PMID- 11017048 TI - Manipulation of human minichromosomes to carry greater than megabase-sized chromosome inserts. AB - For introducing regions of human chromosomes greater than a megabase into cells or animals, we have developed a chromosome-cloning system in which defined regions of human chromosomes can be cloned into a stable human minichromosome vector in homologous recombination-proficient chicken DT40 cells. The stable minichromosome vector allowed a 10 Mb-sized region of the mitotically unstable human chromosome 22 to be stably maintained in mouse embryonic stem (ES) cells, and in mice. Furthermore, we demonstrated functional expression of human genes from the HAC in mice. This study describes a stable cloning and expression system for greater than megabase-sized regions of human chromosomes. PMID- 11017049 TI - Detecting protein analytes that modulate transmembrane movement of a polymer chain within a single protein pore. AB - Here we describe a new type of biosensor element for detecting proteins in solution at nanomolar concentrations. We tethered a 3.4 kDa polyethylene glycol chain at a defined site within the lumen of the transmembrane protein pore formed by staphylococcal alpha-hemolysin. The free end of the polymer was covalently attached to a biotin molecule. On incorporation of the modified pore into a lipid bilayer, the biotinyl group moves from one side of the membrane to the other, and is detected by reversible capture with a mutant streptavidin. The capture events are observed as changes in ionic current passing through single pores in planar bilayers. Accordingly, the modified pore allows detection of a protein analyte at the single-molecule level, facilitating both quantification and identification through a distinctive current signature. The approach has higher time resolution compared with other kinetic measurements, such as those obtained by surface plasmon resonance. PMID- 11017050 TI - Mutation detection by electrocatalysis at DNA-modified electrodes. AB - Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)6]3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and "hot spot" mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection. PMID- 11017052 TI - Discovering novel ligands for macromolecules using X-ray crystallographic screening. AB - The need to decrease the time scale for clinical compound discovery has led to innovations at several stages in the process, including genomics/proteomics for target identification, ultrahigh-throughput screening for lead identification, and structure-based drug design and combinatorial chemistry for lead optimization. A critical juncture in the process is the identification of a proper lead compound, because a poor choice may generate costly difficulties at later stages. Lead compounds are commonly identified from high-throughput screens of large compound libraries, derived from known substrates/inhibitors, or identified in computational prescreeusing X-ray crystal structures. Structural information is often consulted to efficiently optimize leads, but under the current paradigm, such data require preidentification and confirmation of compound binding. Here, we describe a new X-ray crystallography-driven screening technique that combines the steps of lead identification, structural assessment, and optimization. The method is rapid, efficient, and high-throughput, and it results in detailed crystallographic structure information. The utility of the method is demonstrated in the discovery and optimization of a new orally available class of urokinase inhibitors for the treatment of cancer. PMID- 11017051 TI - Field performance of transgenic elite commercial hybrid rice expressing bacillus thuringiensis delta-endotoxin. AB - Here we describe development of transgenic elite rice lines expressing a Bt fusion gene derived from cryIA(b) and cryIA(c) under the control of rice actinI promoter. The lines used in the study were indica CMS restorer line of Minghui 63 and its derived hybrid rice Shanyou 63. The level of Bt fusion protein CryIA(b)/CryIA(c) detected in Minghui 63 (T51-1) plants was 20 ng/mg soluble protein. The Bt Shanyou 63 was field-tested in natural and repeated heavy manual infestation of two lepidopteran insects, leaffolder and yellow stem borer. The transgenic hybrid plants showed high protection against both insect pests without reduced yield. PMID- 11017053 TI - Ligation-mediated PCR for quantitative in vivo footprinting. AB - Ligation-mediated polymerase chain reaction (LM-PCR) is a genomic analysis technique for determination of (1) primary DNA nucleotide sequences (2) cytosine methylation patterns (3) DNA lesion formation and repair, and (4) in vivo protein DNA footprints. However, LM-PCR can be limited by the multiple steps required and the relatively short stretch of sequence (usually <200 bp) that can be analyzed per reaction. We report here a simplified, one-day LM-PCR protocol in which all pipetting steps can be performed by a robotic workstation and which, moreover, provides longer reads (>350 bp) and enhanced signal quality by use of nonradioactive detection and a LI-COR DNA sequencing instrument. Sensitivity comparable to radiolabeling is achieved using oligonucleotide primers that are 5' end labeled with infrared fluorochromes. We showed that the technique could be used for sensitive and reproducible in vivo photofootprinting of the human phosphoglycerate kinase 1 (PGK1) promoter, as well as providing good Maxam Gilbert sequence information. The methods described here should allow high throughput, high-resolution analysis of transcription factor binding and chromatin structure, and also may be useful for sequencing gaps that are refractory to cloning. PMID- 11017055 TI - Considering our genetic legacy PMID- 11017054 TI - Putting your best mode forward. PMID- 11017057 TI - Mass spectrometry and synthesis & sequencing PMID- 11017056 TI - People PMID- 11017058 TI - Targeting zebrafish. PMID- 11017059 TI - Whiffs of selection. PMID- 11017060 TI - A debut for mito-mouse. PMID- 11017061 TI - The genomics gamble. PMID- 11017062 TI - Guilt by association. PMID- 11017063 TI - TOUCHINGbase PMID- 11017064 TI - Prediction of protein interactions: metabolic enzymes are frequently involved in gene fusion. PMID- 11017065 TI - Mutations in GJB6 cause hidrotic ectodermal dysplasia. PMID- 11017066 TI - A view of Neandertal genetic diversity. PMID- 11017068 TI - Am I an insect? PMID- 11017067 TI - Mutations in AXIN2 cause colorectal cancer with defective mismatch repair by activating beta-catenin/TCF signalling. PMID- 11017069 TI - How many diseases does it take to map a gene with SNPs? PMID- 11017070 TI - Choose your target. AB - The technology of modifying endogenous genes has recently been extended from mice to Drosophila and sheep. Concurrently, genomic sequencing is uncovering thousands of previously uncharacterized genes. Armed with today's technologies, what are our best options for delineating the functions of these new genes? PMID- 11017071 TI - Genetic variation in the gene encoding calpain-10 is associated with type 2 diabetes mellitus. AB - Type 2 or non-insulin-dependent diabetes mellitus (NIDDM) is the most common form of diabetes worldwide, affecting approximately 4% of the world's adult population. It is multifactorial in origin with both genetic and environmental factors contributing to its development. A genome-wide screen for type 2 diabetes genes carried out in Mexican Americans localized a susceptibility gene, designated NIDDM1, to chromosome 2. Here we describe the positional cloning of a gene located in the NIDDM1 region that shows association with type 2 diabetes in Mexican Americans and a Northern European population from the Botnia region of Finland. This putative diabetes-susceptibility gene encodes a ubiquitously expressed member of the calpain-like cysteine protease family, calpain-10 (CAPN10). This finding suggests a novel pathway that may contribute to the development of type 2 diabetes. PMID- 11017072 TI - Generation of mice with mitochondrial dysfunction by introducing mouse mtDNA carrying a deletion into zygotes. AB - Mice carrying mitochondrial DNA (mtDNA) with pathogenic mutations would provide a system in which to study how mutant mtDNAs are transmitted and distributed in tissues, resulting in expression of mitochondrial diseases. However, no effective procedures are available for the generation of these mice. Isolation of mouse cells without mtDNA (rho0) enabled us to trap mutant mtDNA that had accumulated in somatic tissues into rho0 cells repopulated with mtDNA (cybrids). We isolated respiration-deficient cybrids with mtDNA carrying a deletion and introduced this mtDNA into fertilized eggs. The mutant mtDNA was transmitted maternally, and its accumulation induced mitochondrial dysfunction in various tissues. Moreover, most of these mice died because of renal failure, suggesting the involvement of mtDNA mutations in the pathogeneses of new diseases. PMID- 11017073 TI - A computational analysis of whole-genome expression data reveals chromosomal domains of gene expression. AB - Chromosome correlation maps display correlations between the expression patterns of genes on the same chromosome. Using these maps, we show here that adjacent pairs of genes, as well as nearby non-adjacent pairs of genes, show correlated expression independent of their orientation. We present specific examples of adjacent pairs with highly correlated expression patterns, in which the promoter of only one of the two genes contains an upstream activating sequence (UAS) known to be associated with that expression pattern. Finally, we show that genes with similar functions tend to occur in adjacent positions along the chromosomes. Our results suggest that, in certain chromosomal expression domains, an UAS can affect the transcription of genes that are not immediately downstream from it. PMID- 11017074 TI - Transposition of maize Ac/Ds transposable elements in the yeast Saccharomyces cerevisiae. AB - Excision by transposons is associated with chromosome breaks; generally, host cell proteins repair this damage, often introducing mutations. Many transposons also use host proteins in the transposition mechanism or in regulation. Transposition in systems lacking host factors that influence the behaviour of these transpositions is useful in determining what those factors are and how they work. In addition, features of transposition and regulation intrinsic to the element itself can be determined. Maize Activator/Dissociation (Ac/Ds) elements transpose in a wide variety of heterologous plants, but their characteristics in these other systems differ from those in maize, including their response to increasing genetic dosage and the types of repair products recovered following excision. Two Arabidopsis thaliana mutants (iae1 and iae2) show increased Ac transposition frequencies. These mutants, and the differences mentioned above, suggest the involvement of host proteins in Ac/Ds activity and potential differences between these proteins among plant species. Here we report that Ac/Ds elements, members of the hAT (hobo, Ac, Tam3) superfamily, transpose in the yeast Saccharomyces cerevisiae, an organism lacking class II ('cut and paste') transposons. This demonstrates that plant-specific proteins are not essential for Ac/Ds transposition. The yeast system is valuable for dissecting the Ac/Ds transposition mechanism and identifying host factors that can influence transposition and the repair of DNA damage induced by Ac/Ds. Mutations caused by Ds excision in yeast suggest formation of a DNA-hairpin intermediate, and reinsertions occur throughout the genome with a frequency similar to that in plants. The high proportion of Ac/Ds reinsertions also makes this system an in vivo mutagenesis and reverse genetics tool in yeast and, presumably, other eukaryotic systems. PMID- 11017075 TI - Large expansion of the ATTCT pentanucleotide repeat in spinocerebellar ataxia type 10. AB - Spinocerebellar ataxia type 10 (SCA10; MIM 603516; refs 1,2) is an autosomal dominant disorder characterized by cerebellar ataxia and seizures. The gene SCA10 maps to a 3.8-cM interval on human chromosome 22q13-qter (refs 1,2). Because several other SCA subtypes show trinucleotide repeat expansions, we examined microsatellites in this region. We found an expansion of a pentanucleotide (ATTCT) repeat in intron 9 of SCA10 in all patients in five Mexican SCA10 families. There was an inverse correlation between the expansion size, up to 22.5 kb larger than the normal allele, and the age of onset (r2=0.34, P=0.018). Analysis of 562 chromosomes from unaffected individuals of various ethnic origins (including 242 chromosomes from Mexican persons) showed a range of 10 to 22 ATTCT repeats with no evidence of expansions. Our data indicate that the new SCA10 intronic ATTCT pentanucleotide repeat in SCA10 patients is unstable and represents the largest microsatellite expansion found so far in the human genome. PMID- 11017076 TI - Genome rearrangement by replication-directed translocation. AB - Gene order in bacteria is poorly conserved during evolution. For example, although many homologous genes are shared by the proteobacteria Escherichia coli, Haemophilus influenzae and Helicobacter pylori, their relative positions are very different in each genome, except local functional clusters such as operons. The complete sequences of the more closely related bacterial genomes, such as pairs of Chlamydia, H. pylori and Mycobacterium species, now allow identification of the processes and mechanisms involved in genome evolution. Here we provide evidence that a substantial proportion of rearrangements in gene order results from recombination sites that are determined by the positions of the replication forks. Our observations suggest that replication has a major role in directing genome evolution. PMID- 11017077 TI - The mouse fidgetin gene defines a new role for AAA family proteins in mammalian development. AB - The mouse mutation fidget arose spontaneously in a heterogeneous albino stock. This mutant mouse is characterized by a side-to-side head-shaking and circling behaviour, due to reduced or absent semicircular canals. Fidget mice also have small eyes, associated with cell-cycle delay and insufficient growth of the retinal neural epithelium, and lower penetrance skeletal abnormalities, including pelvic girdle dysgenesis, skull bone fusions and polydactyly. By positional cloning, we found the gene mutated in fidget mice, fidgetin (Fign), which encodes a new member of the 'meiotic' or subfamily-7 (SF7; ref. 7) group of ATPases associated with diverse cellular activities (AAA proteins). We also discovered two closely related mammalian genes. AAA proteins are molecular chaperones that facilitate a variety of functions, including membrane fusion, proteolysis, peroxisome biogenesis, endosome sorting and meiotic spindle formation, but functions for the SF7 AAA proteins are largely unknown. Fidgetin is the first mutant AAA protein found in a mammalian developmental mutant, thus defining a new role for these proteins in embryonic development. PMID- 11017078 TI - Deletion of a silencer element in Igf2 results in loss of imprinting independent of H19. AB - Igf2 and H19 are closely linked, reciprocally imprinted genes on mouse distal chromosome 7. The paternally expressed Igf2 encodes a potent fetal growth factor and the maternally expressed H19 encodes a non-coding RNA (refs 1,2). Shared endoderm-specific enhancers 3' to H19 are necessary for transcription of the maternal copy of H19 and the paternal copy of Igf2 (ref. 3), a chromatin boundary upstream of H19 preventing access of the enhancers to the maternal Igf2 promoters. Mesoderm-specific control elements have not been identified, and the role of differentially methylated regions (DMRs) in Igf2 has not been addressed. Two DMRs in Igf2 are methylated on the active paternal allele, suggesting that they contain silencers. Here we have deleted the DMR1 region in Igf2. Maternal transmission of the deletion results in biallelic expression of Igf2 in most mesodermally derived tissues without altering H19 imprinting or expression. Paternal or maternal transmission leads to continued postnatal transcription of Igf2, in contrast to the wild-type allele, which is silenced soon after birth. These results reveal a mesodermal silencer, which may be regulated by methylation and which has a major role in H19-independent expression and imprinting control of Igf2. Our results establish a new mechanistic principle for imprinted genes whereby epigenetically regulated silencers interact with enhancers to control expression, and suggest a new mechanism for loss of imprinting (LOI) of Igf2, which may be important in a number of diseases. PMID- 11017079 TI - Nuclear gene OPA1, encoding a mitochondrial dynamin-related protein, is mutated in dominant optic atrophy. AB - Optic atrophy type 1 (OPA1, MIM 165500) is a dominantly inherited optic neuropathy occurring in 1 in 50,000 individuals that features progressive loss in visual acuity leading, in many cases, to legal blindness. Phenotypic variations and loss of retinal ganglion cells, as found in Leber hereditary optic neuropathy (LHON), have suggested possible mitochondrial impairment. The OPA1 gene has been localized to 3q28-q29 (refs 13-19). We describe here a nuclear gene, OPA1, that maps within the candidate region and encodes a dynamin-related protein localized to mitochondria. We found four different OPA1 mutations, including frameshift and missense mutations, to segregate with the disease, demonstrating a role for mitochondria in retinal ganglion cell pathophysiology. PMID- 11017080 TI - OPA1, encoding a dynamin-related GTPase, is mutated in autosomal dominant optic atrophy linked to chromosome 3q28. AB - Autosomal dominant optic atrophy (ADOA) is the most prevalent hereditary optic neuropathy resulting in progressive loss of visual acuity, centrocoecal scotoma and bilateral temporal atrophy of the optic nerve with an onset within the first two decades of life. The predominant locus for this disorder (OPA1; MIM 165500) has been mapped to a 1.4-cM interval on chromosome 3q28-q29 flanked by markers D3S3669 and D3S3562 (ref. 3). We established a PAC contig covering the entire OPA1 candidate region of approximately 1 Mb and a sequence skimming approach allowed us to identify a gene encoding a polypeptide of 960 amino acids with homology to dynamin-related GTPases. The gene comprises 28 coding exons and spans more than 40 kb of genomic sequence. Upon sequence analysis, we identified mutations in seven independent families with ADOA. The mutations include missense and nonsense alterations, deletions and insertions, which all segregate with the disease in these families. Because most mutations probably represent null alleles, dominant inheritance of the disease may result from haploinsufficiency of OPA1. OPA1 is widely expressed and is most abundant in the retina. The presence of consensus signal peptide sequences suggests that the product of the gene OPA1 is targeted to mitochondria and may exert its function in mitochondrial biogenesis and stabilization of mitochondrial membrane integrity. PMID- 11017081 TI - Effective targeted gene 'knockdown' in zebrafish. AB - The sequencing of the zebrafish genome should be completed by the end of 2002. Direct assignment of function on the basis of this information would be facilitated by the development of a rapid, targeted 'knockdown' technology in this model vertebrate. We show here that antisense, morpholino-modified oligonucleotides (morpholinos) are effective and specific translational inhibitors in zebrafish. We generated phenocopies of mutations of the genes no tail (ref. 2), chordin (ref. 3), one-eyed-pinhead (ref. 4), nacre (ref. 5) and sparse (ref. 6), removing gene function from maternal through post-segmentation and organogenesis developmental stages. We blocked expression from a ubiquitous green fluorescent protein (GFP) transgene, showing that, unlike tissue-restricted limitations found with RNA-based interference in the nematode, all zebrafish cells readily respond to this technique. We also developed also morpholino-based zebrafish models of human disease. Morpholinos targeted to the uroporphyrinogen decarboxylase gene result in embryos with hepatoerythropoietic porphyria. We also used morpholinos for the determination of new gene functions. We showed that embryos with reduced sonic hedgehog (ref. 9) signalling and reduced tiggy-winkle hedgehog (ref. 10) function exhibit partial cyclopia and other specific midline abnormalities, providing a zebrafish genetic model for the common human disorder holoprosencephaly. Conserved vertebrate processes and diseases are now amenable to a systematic, in vivo, reverse-genetic paradigm using zebrafish embryos. PMID- 11017082 TI - Dichotomy of single-nucleotide polymorphism haplotypes in olfactory receptor genes and pseudogenes. AB - Substantial efforts are focused on identifying single-nucleotide polymorphisms (SNPs) throughout the human genome, particularly in coding regions (cSNPs), for both linkage disequilibrium and association studies. Less attention, however, has been directed to the clarification of evolutionary processes that are responsible for the variability in nucleotide diversity among different regions of the genome. We report here the population sequence diversity of genomic segments within a 450-kb cluster of olfactory receptor (OR) genes on human chromosome 17. We found a dichotomy in the pattern of nucleotide diversity between OR pseudogenes and introns on the one hand and the closely interspersed intact genes on the other. We suggest that weak positive selection is responsible for the observed patterns of genetic variation. This is inferred from a lower ratio of polymorphism to divergence in genes compared with pseudogenes or introns, high non-synonymous substitution rates in OR genes, and a small but significant overall reduction in variability in the entire OR gene cluster compared with other genomic regions. The dichotomy among functionally different segments within a short genomic distance requires high recombination rates within this OR cluster. Our work demonstrates the impact of weak positive selection on human nucleotide diversity, and has implications for the evolution of the olfactory repertoire. PMID- 11017083 TI - Human-mouse genome comparisons to locate regulatory sites. AB - Elucidating the human transcriptional regulatory network is a challenge of the post-genomic era. Technical progress so far is impressive, including detailed understanding of regulatory mechanisms for at least a few genes in multicellular organisms, rapid and precise localization of regulatory regions within extensive regions of DNA by means of cross-species comparison, and de novo determination of transcription-factor binding specificities from large-scale yeast expression data. Here we address two problems involved in extending these results to the human genome: first, it has been unclear how many model organism genomes will be needed to delineate most regulatory regions; and second, the discovery of transcription-factor binding sites (response elements) from expression data has not yet been generalized from single-celled organisms to multicellular organisms. We found that 98% (74/75) of experimentally defined sequence-specific binding sites of skeletal-muscle-specific transcription factors are confined to the 19% of human sequences that are most conserved in the orthologous rodent sequences. Also we found that in using this restriction, the binding specificities of all three major muscle-specific transcription factors (MYF, SRF and MEF2) can be computationally identified. PMID- 11017084 TI - Loss of Gcn5l2 leads to increased apoptosis and mesodermal defects during mouse development. AB - Histone acetyltransferases regulate transcription, but little is known about the role of these enzymes in developmental processes. Gcn5 (encoded by Gcn5l2) and Pcaf, mouse histone acetyltransferases, share similar sequences and enzymatic activities. Both interact with p300 and CBP (encoded by Ep300 and Crebbp, respectively), two other histone acetyltransferases that integrate multiple signalling pathways. Pcaf is thought to participate in many of the cellular processes regulated by p300/CBP (refs 2-8), but the functions of Gcn5 are unknown in mammalian cells. Here we show that the gene Pcaf is dispensable in mice. In contrast, Gcn5l2-null embryos die during embryogenesis. These embryos develop normally to 7.5 days post coitum (d.p.c.), but their growth is severely retarded by 8.5 d.p.c. and they fail to form dorsal mesoderm lineages, including chordamesoderm and paraxial mesoderm. Differentiation of extra-embryonic and cardiac mesoderm seems to be unaffected. Loss of the dorsal mesoderm lineages is due to a high incidence of apoptosis in the Gcn5l2 mutants that begins before the onset of morphological abnormality. Embryos null for both Gcn5l2 and Pcaf show even more severe defects, indicating that these histone acetyltransferases have overlapping functions during embryogenesis. Our studies are the first to demonstrate that specific acetyltransferases are required for cell survival and mesoderm formation during mammalian development. PMID- 11017085 TI - Genome-wide analysis of single-nucleotide polymorphisms in human expressed sequences. AB - Single-nucleotide polymorphisms (SNPs) have been explored as a high-resolution marker set for accelerating the mapping of disease genes. Here we report 48,196 candidate SNPs detected by statistical analysis of human expressed sequence tags (ESTs), associated primarily with coding regions of genes. We used Bayesian inference to weigh evidence for true polymorphism versus sequencing error, misalignment or ambiguity, misclustering or chimaeric EST sequences, assessing data such as raw chromatogram height, sharpness, overlap and spacing, sequencing error rates, context-sensitivity and cDNA library origin. Three separate validations-comparison with 54 genes screened for SNPs independently, verification of HLA-A polymorphisms and restriction fragment length polymorphism (RFLP) testing-verified 70%, 89% and 71% of our predicted SNPs, respectively. Our method detects tenfold more true HLA-A SNPs than previous analyses of the EST data. We found SNPs in a large fraction of known disease genes, including some disease-causing mutations (for example, the HbS sickle-cell mutation). Our comprehensive analysis of human coding region polymorphism provides a public resource for mapping of disease genes (available at http://www.bioinformatics.ucla.edu/snp). PMID- 11017086 TI - Macular corneal dystrophy type I and type II are caused by distinct mutations in a new sulphotransferase gene. AB - Macular corneal dystrophy (MCD; MIM 217800) is an autosomal recessive hereditary disease in which progressive punctate opacities in the cornea result in bilateral loss of vision, eventually necessitating corneal transplantation. MCD is classified into two subtypes, type I and type II, defined by the respective absence and presence of sulphated keratan sulphate in the patient serum, although both types have clinically indistinguishable phenotypes. The gene responsible for MCD type I has been mapped to chromosome 16q22, and that responsible for MCD type II may involve the same locus. Here we identify a new carbohydrate sulphotransferase gene (CHST6), encoding an enzyme designated corneal N acetylglucosamine-6-sulphotransferase (C-GlcNAc6ST), within the critical region of MCD type I. In MCD type I, we identified several mutations that may lead to inactivation of C-GlcNAc6ST within the coding region of CHST6. In MCD type II, we found large deletions and/or replacements caused by homologous recombination in the upstream region of CHST6. In situ hybridization analysis did not detect CHST6 transcripts in corneal epithelium in an MCD type II patient, suggesting that the mutations found in type II lead to loss of cornea-specific expression of CHST6. PMID- 11017087 TI - Biochemical defects in ABCR protein variants associated with human retinopathies. AB - Mutations in the gene encoding ABCR (ABCA4), a photoreceptor-specific ATP-binding cassette (ABC) transporter, are responsible for autosomal recessive Stargardt disease (STGD), an early onset macular degeneration, and some forms of autosomal recessive cone-rod dystrophy and autosomal recessive retinitis pigmentosa. Heterozygosity for ABCA4 mutations may also represent a risk factor for age related macular degeneration (AMD), although this idea is controversial. An ongoing challenge in the analysis of ABCA4-based retinopathies arises from the observation that most of the ABCA4 sequence variants identified so far are missense mutations that are rare in both patient and control populations. With the current sample size of most sequence variants, one cannot determine statistically whether a particular sequence variant is pathogenic or neutral. A related challenge is to determine the degree to which each pathogenic variant impairs ABCR function, as genotype-phenotype analyses indicate that age of onset and disease severity correlate with different ABCA4 alleles. To address these questions, we performed a functional analysis of human ABCR and its variants. These experiments reveal a wide spectrum of biochemical defects in these variants and provide insight into the transport mechanism of ABCR. PMID- 11017088 TI - Mutations in ARHGEF6, encoding a guanine nucleotide exchange factor for Rho GTPases, in patients with X-linked mental retardation. AB - X-linked forms of mental retardation (XLMR) include a variety of different disorders and may account for up to 25% of all inherited cases of mental retardation. So far, seven X-chromosomal genes mutated in nonspecific mental retardation (MRX) have been identified: FMR2, GDI1, RPS6KA3, IL1RAPL, TM4SF2, OPHN1 and PAK3 (refs 2-9). The products of the latter two have been implicated in regulation of neural plasticity by controlling the activity of small GTPases of the Rho family. Here we report the identification of a new MRX gene, ARHGEF6 (also known as alphaPIX or Cool-2), encoding a protein with homology to guanine nucleotide exchange factors for Rho GTPases (Rho GEF). Molecular analysis of a reciprocal X/21 translocation in a male with mental retardation showed that this gene in Xq26 was disrupted by the rearrangement. Mutation screening of 119 patients with nonspecific mental retardation revealed a mutation in the first intron of ARHGEF6 (IVS1-11T-->C) in all affected males in a large Dutch family. The mutation resulted in preferential skipping of exon 2, predicting a protein lacking 28 amino acids. ARHGEF6 is the eighth MRX gene identified so far and the third such gene to encode a protein that interacts with Rho GTPases. PMID- 11017089 TI - In cooperation we trust. PMID- 11017090 TI - Type 1 IFNs in human versus mouse. PMID- 11017091 TI - AIDS in Africa: a disaster no longer waiting to happen. PMID- 11017092 TI - If at first you don't succeed. PMID- 11017093 TI - Flexibility and cooperation among dendritic cells. PMID- 11017094 TI - T cells: getting a GRP on Ras. PMID- 11017095 TI - Incompatible differences: view of an allogeneic pMHC-TCR complex. PMID- 11017096 TI - Signal transduction on the fly. PMID- 11017097 TI - Immunology highlights from the recent literature. PMID- 11017098 TI - T cell depletion in HIV-1 infection: how CD4+ T cells go out of stock. AB - HIV-1 infection is characterized by a gradual loss of CD4+ T cells and progressive immune deficiency that leads to opportunistic infections, otherwise rare malignancies and ultimately death. Extensive research over the past two decades has increased our insight into the pathogenic mechanisms underlying these features of HIV-1 infection. Here, we will give a brief overview of the most recent findings and present a model that fits most of the relevant aspects of HIV 1 infection as known. We hypothesize that HIV-1 infection depletes T cell supplies (which are not replaced because of low and static thymic function) by direct infection and killing of cells and through hyperactivation of the immune system. PMID- 11017099 TI - Crystal structure of a T cell receptor bound to an allogeneic MHC molecule. AB - Many T cell receptors (TCRs) that are selected to respond to foreign peptide antigens bound to self major histocompatibility complex (MHC) molecules are also reactive with allelic variants of self-MHC molecules. This property, termed alloreactivity, causes graft rejection and graft-versus-host disease. The structural features of alloreactivity have yet to be defined. We now present a basis for this cross-reactivity, elucidated by the crystal structure of a complex involving the BM3.3 TCR and a naturally processed octapeptide bound to the H-2Kb allogeneic MHC class I molecule. A distinguishing feature of this complex is that the eleven-residue-long complementarity-determining region 3 (CDR3) found in the BM3.3 TCR alpha chain folds away from the peptide binding groove and makes no contact with the bound peptide, the latter being exclusively contacted by the BM3.3 CDR3 beta. Our results formally establish that peptide-specific, alloreactive TCRs interact with allo-MHC in a register similar to the one they use to contact self-MHC molecules. PMID- 11017100 TI - A transmembrane CXC chemokine is a ligand for HIV-coreceptor Bonzo. AB - We describe a protein with the hallmarks of a chemokine, designated CXCL16, that is made by dendritic cells (DCs) in lymphoid organ T cell zones and by cells in the splenic red pulp. CXCL16 contains a transmembrane domain and both membrane bound and soluble forms are produced. Naive CD8 T cells, natural killer T cells and a subset of memory CD4 T cells bind CXCL16, and activated T cells migrated chemotactically to the soluble chemokine. By expression cloning, Bonzo (also known as STRL33 and TYMSTR) was identified as a CXCL16 receptor. CXCL16 may function in promoting interactions between DCs and CD8 T cells and in guiding T cell movements in the splenic red pulp. CXCL16 was also found in the thymic medulla and in some nonlymphoid tissues, indicating roles in thymocyte development and effector T cell trafficking. PMID- 11017101 TI - Plasmacytoid dendritic cells activated by influenza virus and CD40L drive a potent TH1 polarization. AB - Plasmacytoid dendritic cells (PDCs) are a subset of dendritic cells present in human blood and inflamed lymph nodes. Here we show that blood PDCs, when stimulated with influenza virus and CD40L in vitro, undergo a maturation process characterized by up-regulation of major histocompatibility complex proteins and adhesion and costimulatory molecules. In addition, PDCs down-regulate CXCR3 and L selectin, which mediate migration and homing of these cells into the lymph node. Mature PDCs efficiently stimulate T cells and drive a potent TH1 polarization in vitro, which is mediated by the synergistic effect of interleukin 12 and type 1 interferon. In vivo, mature PDCs are found in secondary lymphoid organs, where they represent the principal source of type 1 interferon during inflammation. Thus, PDCs probably participate in antiviral and pro-inflammatory responses, rather than in TH2 polarization and tolerance induction. PMID- 11017102 TI - Kinetics of dendritic cell activation: impact on priming of TH1, TH2 and nonpolarized T cells. AB - To prime immune responses, dendritic cells (DCs) need to be activated to acquire T cell stimulatory capacity. Although some stimuli trigger interleukin 12 (IL-12) production that leads to T helper cell type I (TH1) polarization, others fail to do so and favor TH2 polarization. We show that after activation by lipopolysaccharide, DCs produced IL-12 only transiently and became refractory to further stimulation. The exhaustion of cytokine production impacted the T cell polarizing process. Soon after stimulation DCs primed strong TH1 responses, whereas at later time points the same cells preferentially primed TH2 and nonpolarized T cells. These findings indicate that during an immune response, T cell priming conditions may change in the lymph nodes, suggesting another mechanism for the regulation of effector and memory T cells. PMID- 11017103 TI - RasGRP is essential for mouse thymocyte differentiation and TCR signaling. AB - The Ras signaling pathway plays a critical role in thymopoiesis and T cell activation, but the mechanism of Ras regulation is controversial. At least one mode of Ras regulation in T cells involves the messenger diacylglycerol (DAG). RasGRP, a Ras activator with a DAG-binding C1 domain, is expressed in T cells and thymocytes. Here we show that thymi of RasGRP-null mutant mice have approximately normal numbers of immature thymocytes but a marked deficiency of mature, single positive (CD4+CD8- and CD4-CD8+) thymocytes. In Ras signaling and proliferation assays, mutant thymocytes showed a complete lack of response to DAG analogs or T cell receptor (TCR) stimulation by antibodies. Thus, TCR and DAG are linked through RasGRP to Ras signaling. PMID- 11017104 TI - The 21- and 23-kD forms of TCR zeta are generated by specific ITAM phosphorylations. AB - The T cell receptor (TCR) zeta subunit contains three immunoreceptor tyrosine based activation motifs (ITAMs) that translate effective extracellular ligand binding into intracellular signals by becoming phosphorylated into 21- and 23-kD forms. We report here that the 21-kD form of TCR zeta is generated by phosphorylation of the tyrosines in the second and third ITAMs, whereas the 23-kD form is formed by the additional phosphorylation of the membrane-proximal ITAM tyrosines. The stable formation of the 21- and 23-kD species requires the binding of the tandem SH2 domains of ZAP-70. We also report that TCR-mediated signaling processes can proceed independently of either the 21- or 23-kD species of TCR zeta. PMID- 11017105 TI - CD4+ T cell survival is not directly linked to self-MHC-induced TCR signaling. AB - T cell receptor (TCR) signaling triggered by recognition of self-major histocompatibility complex (MHC) ligands has been proposed to maintain the viability of naive T cells and to provoke their proliferation in T cell-deficient hosts. Consistent with this, the partially phosphorylated state of TCR zeta chains in naive CD4+ and CD8+ T cells in vivo was found to be actively maintained by TCR interactions with specific peptide-containing MHC molecules. TCR ligand dependent phosphorylation of TCR zeta was lost within one day of cell transfer into MHC-deficient hosts, yet the survival of transferred CD4+ lymphocytes was the same in recipients with or without MHC class II expression for one month. Thus, despite clear evidence for TCR signaling in nonactivated naive T cells, these data argue against the concept that such signaling plays a predominant role in determining lymphocyte lifespan. PMID- 11017106 TI - Receptor editing in developing T cells. AB - A central tenet of T cell development postulates that if a developing thymocyte encounters self-antigen, it is induced to die via apoptosis, thereby protecting the organism from autoreactive T cells. We created transgenic mice that expressed a peptide antigen in the cortical epithelial cells of the thymus. This did not, however, result in deletion of specific T cells. Instead, antigen presentation by epithelial cells caused T cell receptor (TCR) internalization and increased gene rearrangement at the endogenous TCR alpha locus, or receptor editing. This editing mechanism in immature T cells parallels that which occurs in immature B cells, and has important implications for understanding positive and negative selection signaling in the thymus, and the limits of self-tolerance. PMID- 11017107 TI - Role of Drosophila IKK gamma in a toll-independent antibacterial immune response. AB - We have generated, by ethylmethane sulfonate mutagenesis, loss-of-function mutants in the Drosophila homolog of the mammalian I-kappa B kinase (IKK) complex component IKK gamma (also called NEMO). Our data show that Drosophila IKK gamma is required for the Relish-dependent immune induction of the genes encoding antibacterial peptides and for resistance to infections by Escherichia coli. However, it is not required for the Toll-DIF-dependent antifungal host defense. The results indicate distinct control mechanisms of the Rel-like transactivators DIF and Relish in the Drosophila innate immune response and show that Drosophila Toll does not signal through a IKK gamma-dependent signaling complex. Thus, in contrast to the vertebrate inflammatory response, IKK gamma is required for the activation of only one immune signaling pathway in Drosophila. PMID- 11017108 TI - Control of pre-T cell proliferation and differentiation by the GTPase Rac-I. AB - The GTPase Rac-I has the potential for pleiotropic functions due to its ability to interact with multiple effectors. Here, activation of Rac-I is shown to potently regulate pre-T cell differentiation and proliferation at the point of T cell antigen receptor (TCR) beta selection. An activated Rac-I effector domain mutant that restricts signaling to particular actions on actin dynamics can drive pre-T cell differentiation. Rac-I activation cannot fully substitute for the pre TCR complex but can fully correct defects in pre-T cell development in mice lacking the adapter molecule Vav-1. The present study identifies the subset of Rac-I responses that mediate Vav-1 action as critical regulators of TCR beta selection. PMID- 11017109 TI - DC-SIGN-ICAM-2 interaction mediates dendritic cell trafficking. AB - Dendritic cells (DCs) are recruited from blood into tissues to patrol for foreign antigens. After antigen uptake and processing, DCs migrate to the secondary lymphoid organs to initiate immune responses. We now show that DC-SIGN, a DC specific C-type lectin, supports tethering and rolling of DC-SIGN-positive cells on the vascular ligand ICAM-2 under shear flow, a prerequisite for emigration from blood. The DC-SIGN-ICAM-2 interaction regulates chemokine-induced transmigration of DCs across both resting and activated endothelium. Thus, DC SIGN is central to the unusual trafficking capacity of DCs, further supported by the expression of DC-SIGN on precursors in blood and on immature and mature DCs in both peripheral and lymphoid tissues. PMID- 11017110 TI - Untangling huntingtin's mysteries. PMID- 11017111 TI - LMP2 expression and proteasome activity in NOD mice. PMID- 11017112 TI - LMP2 expression and proteasome activity in NOD mice. PMID- 11017113 TI - Reply to 'LMP2 expression and proteasome activity in NOD mice' PMID- 11017114 TI - Results make a monkey of OPV-AIDS theory. PMID- 11017115 TI - While US scientists push for derivation. PMID- 11017116 TI - Europe asks UK to reject cloning approval. PMID- 11017117 TI - Pope influences biomedical scientists. PMID- 11017119 TI - Oxford scientists defect to imperial PMID- 11017118 TI - Pediatric study of medicines becomes mandatory. PMID- 11017120 TI - Research legacy of suicide surgeon. PMID- 11017121 TI - Bioinformaticians develop new data mining tools. PMID- 11017122 TI - Can italy catch up in genomics research? PMID- 11017123 TI - NIH 'glues' cell biologists together. PMID- 11017124 TI - Teaching hospitals to share tissue with industry. PMID- 11017125 TI - Basic Medical Research Award. The ubiquitin system. PMID- 11017126 TI - Clinical Medical Research Award. Hepatitis C virus and eliminating post transfusion hepatitis. PMID- 11017127 TI - Special Achievement in Medical Science Award. From cell physiology to cell physiology. PMID- 11017128 TI - The door in the dream: conversations with eminent women in science PMID- 11017129 TI - Driving mr. Albert: A trip across america with Einstein's brain PMID- 11017130 TI - Learning old tricks from new viruses. PMID- 11017131 TI - Uncoupling diet and diabetes. PMID- 11017132 TI - Containing HIV after infection. PMID- 11017133 TI - Skin stem cells - a hairy issue. PMID- 11017134 TI - Breaking up receptor alliances: the parting of CD3 and CD4. PMID- 11017135 TI - Specific immunotherapy: one size does not fit all. PMID- 11017136 TI - Protecting the female germ line from cancer therapy. PMID- 11017137 TI - VEGF gene therapy: stimulating angiogenesis or angioma-genesis? PMID- 11017138 TI - 'Orphans' meet cholesterol. PMID- 11017140 TI - Research news PMID- 11017139 TI - A super way to kill cancer cells? PMID- 11017141 TI - Oocyte apoptosis is suppressed by disruption of the acid sphingomyelinase gene or by sphingosine-1-phosphate therapy. AB - The time at which ovarian failure (menopause) occurs in females is determined by the size of the oocyte reserve provided at birth, as well as by the rate at which this endowment is depleted throughout post-natal life. Here we show that disruption of the gene for acid sphingomyelinase in female mice suppressed the normal apoptotic deletion of fetal oocytes, leading to neonatal ovarian hyperplasia. Ex vivo, oocytes lacking the gene for acid sphingomyelinase or wild type oocytes treated with sphingosine-1-phosphate resisted developmental apoptosis and apoptosis induced by anti-cancer therapy, confirming cell autonomy of the death defect. Moreover, radiation-induced oocyte loss in adult wild-type female mice, the event that drives premature ovarian failure and infertility in female cancer patients, was completely prevented by in vivo therapy with sphingosine-1-phosphate. Thus, the sphingomyelin pathway regulates developmental death of oocytes, and sphingosine-1-phosphate provides a new approach to preserve ovarian function in vivo. PMID- 11017142 TI - Skeletal muscle respiratory uncoupling prevents diet-induced obesity and insulin resistance in mice. AB - To determine whether uncoupling respiration from oxidative phosphorylation in skeletal muscle is a suitable treatment for obesity and type 2 diabetes, we generated transgenic mice expressing the mitochondrial uncoupling protein (Ucp) in skeletal muscle. Skeletal muscle oxygen consumption was 98% higher in Ucp-L mice (with low expression) and 246% higher in Ucp-H mice (with high expression) than in wild-type mice. Ucp mice fed a chow diet had the same food intake as wild type mice, but weighed less and had lower levels of glucose and triglycerides and better glucose tolerance than did control mice. Ucp-L mice were resistant to obesity induced by two different high-fat diets. Ucp-L mice fed a high-fat diet had less adiposity, lower levels of glucose, insulin and cholesterol, and an increased metabolic rate at rest and with exercise. They were also more responsive to insulin, and had enhanced glucose transport in skeletal muscle in the setting of increased muscle triglyceride content. These data suggest that manipulating respiratory uncoupling in muscle is a viable treatment for obesity and its metabolic sequelae. PMID- 11017143 TI - The latent nuclear antigen of Kaposi sarcoma-associated herpesvirus targets the retinoblastoma-E2F pathway and with the oncogene Hras transforms primary rat cells. AB - Kaposi sarcoma-associated herpesvirus (KSHV) is involved in the etiopathogenesis of Kaposi sar-coma and certain lymphoproliferative disorders. Open reading frame (ORF) 73 encodes the main immunogenic latent nuclear antigen (LNA-1) of KSHV. LNA 1 maintains the KSHV episome and tethers the viral genome to chromatin during mitosis. In addition, LNA-1 interacts with p53 and represses its transcriptional activity. Here we show that LNA-1 also interacts with the retinoblastoma protein. LNA-1 transactivated an artificial promoter carrying the cell cycle transcription factor E2F DNA-binding sequences and also upregulated the cyclin E (CCNEI) promoter, but not the B-myb (MYBL2) promoter. LNA-1 overcame the flat-cell phenotype induced by retinoblastoma protein in Saos2 cells. In cooperation with the cellular oncogene Harvey rat sarcoma viral oncogene homolog (Hras), LNA-1 transformed primary rat embryo fibroblasts and rendered them tumorigenic. These findings indicate that LNA-1 acts as a transcription co-factor and may contribute to KSHV-induced oncogenesis by targeting the retinoblastoma protein-E2F transcriptional regulatory pathway. PMID- 11017144 TI - Loss of p14ARF in tumor cells facilitates replication of the adenovirus mutant dl1520 (ONYX-015). AB - The adenovirus mutant dl1520 (ONYX-015) does not express the E1B-55K protein that binds and inactivates p53. This virus replicates in tumor cells with mutant p53, but not in normal cells with functional p53. Although intra-tumoral injection of dl1520 shows promising responses in patients with solid tumors, previous in vitro studies have not established a close correlation between p53 status and dl1520 replication. Here we identify loss of p14ARF as a mechanism that allows dl1520 replication in tumor cells retaining wild-type p53. We demonstrate that the re introduction of p14ARF into tumor cells with wild-type p53 suppresses replication of dl1520 in a p53-dependent manner. Our study supports the therapeutic use of dl1520 in tumors with lesions within the p53 pathway other than mutation of p53. PMID- 11017145 TI - An adenovirus E1A mutant that demonstrates potent and selective systemic anti tumoral efficacy. AB - Replication-selective oncolytic viruses constitute a rapidly evolving and new treatment platform for cancer. Gene-deleted viruses have been engineered for tumor selectivity, but these gene deletions also reduce the anti-cancer potency of the viruses. We have identified an E1A mutant adenovirus, dl922-947, that replicates in and lyses a broad range of cancer cells with abnormalities in cell cycle checkpoints. This mutant demonstrated reduced S-phase induction and replication in non-proliferating normal cells, and superior in vivo potency relative to other gene-deleted adenoviruses. In some cancers, its potency was superior to even wild-type adenovirus. Intravenous administration reduced the incidence of metastases in a breast tumor xenograft model. dl922-947 holds promise as a potent, replication-selective virus for the local and systemic treatment of cancer. PMID- 11017146 TI - Viremia control following antiretroviral treatment and therapeutic immunization during primary SIV251 infection of macaques. AB - Prolonged antiretroviral therapy (ART) is not likely to eradicate human immunodeficiency virus type I (HIV-I) infection. Here we explore the effect of therapeutic immunization in the context of ART during primary infection using the simian immunodeficiency virus (SIV251) macaque model. Vaccination of rhesus macaques with the highly attenuated poxvirus-based NYVAC-SIV vaccine expressing structural genes elicited vigorous virus-specific CD4 + and CD8+ T cell responses in macaques that responded effectively to ART. Following discontinuation of a six month ART regimen, viral rebound occurred in most animals, but was transient in six of eight vaccinated animals. Viral rebound was also transient in four of seven mock-vaccinated control animals. These data establish the importance of antiretroviral treatment during primary infection and demonstrate that virus specific immune responses in the infected host can be expanded by therapeutic immunization. PMID- 11017147 TI - Secretory leukocyte protease inhibitor mediates non-redundant functions necessary for normal wound healing. AB - Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor with anti-microbial properties found in mucosal fluids. It is expressed during cutaneous wound healing. Impaired healing states are characterized by excessive proteolysis and often bacterial infection, leading to the hypothesis that SLPI may have a role in this process. We have generated mice null for the gene encoding SLPI (Slpi), which show impaired cutaneous wound healing with increased inflammation and elastase activity. The altered inflammatory profile involves enhanced activation of local TGF-beta in Slpi-null mice. We propose that SLPI is a pivotal endogenous factor necessary for optimal wound healing. PMID- 11017148 TI - Dendritic cells genetically modified to express CD40 ligand and pulsed with antigen can initiate antigen-specific humoral immunity independent of CD4+ T cells. AB - We have investigated whether dendritic cells genetically modified to express CD40 ligand and pulsed with antigen can trigger B cells to produce antigen-specific antibodies without CD4+ T-cell help. Dendritic cells modified with a recombinant adenovirus vector to express CD40 ligand and pulsed with heat-killed Pseudomonas induced naive B cells to produce antibodies against Pseudomonas in the absence of CD4+ T cells in vitro, initiated Pseudomonas-specific humoral immune responses in vivo in wild-type and CD4-/- mice, and protected immunized wild-type and CD4-/-, but not B-cell -/- mice, from lethal intrapulmonary challenge with Pseudomonas. Thus, genetic modification of dendritic cells with CD40 ligand enables them to present a complex mixture of microbial antigens and establish CD4+ T cell independent, B cell-mediated protective immunity against a specific microbe. PMID- 11017149 TI - Immunotherapy of tumors with xenogeneic endothelial cells as a vaccine. AB - The breaking of immune tolerance against autologous angiogenic endothelial cells should be a useful approach for cancer therapy. Here we show that immunotherapy of tumors using fixed xenogeneic whole endothelial cells as a vaccine was effective in affording protection from tumor growth, inducing regression of established tumors and prolonging survival of tumor-bearing mice. Furthermore, autoreactive immunity targeting to microvessels in solid tumors was induced and was probably responsible for the anti-tumor activity. These observations may provide a new vaccine strategy for cancer therapy through the induction of an autoimmune response against the tumor endothelium in a cross-reaction. PMID- 11017150 TI - Encephalitogenic potential of the myelin basic protein peptide (amino acids 83 99) in multiple sclerosis: results of a phase II clinical trial with an altered peptide ligand. AB - Myelin-specific T lymphocytes are considered essential in the pathogenesis of multiple sclerosis. The myelin basic protein peptide (a.a. 83-99) represents one candidate antigen; therefore, it was chosen to design an altered peptide ligand, CGP77116, for specific immunotherapy of multiple sclerosis. A magnetic resonance imaging-controlled phase II clinical trial with this altered peptide ligand documented that it was poorly tolerated at the dose tested, and the trial had therefore to be halted. Improvement or worsening of clinical or magnetic resonance imaging parameters could not be demonstrated in this small group of individuals because of the short treatment duration. Three patients developed exacerbations of multiple sclerosis, and in two this could be linked to altered peptide ligand treatment by immunological studies demonstrating the encephalitogenic potential of the myelin basic protein peptide (a.a. 83-99) in a subgroup of patients. These data raise important considerations for the use of specific immunotherapies in general. PMID- 11017152 TI - Systemic sclerosis immunoglobulin G autoantibodies bind the human cytomegalovirus late protein UL94 and induce apoptosis in human endothelial cells. AB - Systemic sclerosis is an autoimmune disease characterized by immunological and vascular abnormalities. Autoantibodies against intracellular antigens are associated with particular clinical features of the disease, whereas autoantibodies against cell surface antigens may be pathogenic by inducing endothelial cell damage, considered the primary event in the pathogenesis of the disease. Latent human cytomegalovirus infection may contribute to progression of systemic sclerosis through its ability to infect endothelial cells; however, direct links between human cytomegalovirus infection and systemic sclerosis are still lacking. Molecular mimicry is one of the mechanisms that account for the link between infection and autoimmunity. Here we have identified an immunodominant peptide using systemic sclerosis serum screening of a random peptide library; such peptide shares homology with autoantigens and with the human cytomegalovirus late protein UL94 (ref. 9). Immunoglobulin G antibodies against the peptide affinity-purified from the sera of patients with systemic sclerosis specifically recognized the viral product and autoantigens; moreover, such antibodies induced endothelial cell apoptosis through specific interaction with the cell surface integrin-NAG-2 protein complex. Our results provide evidence that antibodies against human cytomegalovirus cause apoptosis of endothelial cells, considered the initial pathogenic event of systemic sclerosis, and indicate a previously unknown mechanism for the etiological link between human cytomegalovirus infection and autoimmunity. PMID- 11017151 TI - Induction of a non-encephalitogenic type 2 T helper-cell autoimmune response in multiple sclerosis after administration of an altered peptide ligand in a placebo controlled, randomized phase II trial. The Altered Peptide Ligand in Relapsing MS Study Group. AB - In this 'double-blind', randomized, placebo-controlled phase II trial, we compared an altered peptide ligand of myelin basic protein with placebo, evaluating their safety and influence on magnetic resonance imaging in relapsing remitting multiple sclerosis. A safety board suspended the trial because of hypersensitivity reactions in 9% of the patients. There were no increases in either clinical relapses or in new enhancing lesions in any patient, even those with hypersensitivity reactions. Secondary analysis of those patients completing the study showed that the volume and number of enhancing lesions were reduced at a dose of 5 mg. There was also a regulatory type 2 T helper-cell response to altered peptide ligand that cross-reacted with the native peptide. PMID- 11017153 TI - Epidermal immunization by a needle-free powder delivery technology: immunogenicity of influenza vaccine and protection in mice. PMID- 11017154 TI - On the market PMID- 11017156 TI - Reply to 'One motor cortex, two different views' PMID- 11017155 TI - Stem cells show their muscle. PMID- 11017157 TI - One motor cortex, two different views. PMID- 11017158 TI - One motor cortex, two different views. PMID- 11017159 TI - Reply to 'One motor cortex, two different views' PMID- 11017160 TI - Reply to 'One motor cortex, two different views' PMID- 11017162 TI - Reply to "What', 'where' and 'how' in auditory cortex' PMID- 11017161 TI - 'What', 'where' and 'how' in auditory cortex. PMID- 11017163 TI - Achieving neuronal patterning by repression. PMID- 11017164 TI - SNARE-ing neurotransmitter transporters. PMID- 11017165 TI - CPG15 and the dynamics of retinotectal synapses. PMID- 11017167 TI - The emperor has some clothes PMID- 11017166 TI - Early detection of Alzheimer's disease: an fMRI marker for people at risk? PMID- 11017168 TI - Corticosteroid actions in hippocampus require DNA binding of glucocorticoid receptor homodimers. AB - Glucocorticoids are secreted from the adrenal gland in very high amounts after stress. In the brain, these stress hormones potently modulate ionic currents, monoaminergic transmission, synaptic plasticity and cellular viability, most notably in the hippocampus where corticosteroid receptors are highly enriched. Here we show that at least some of these actions require DNA binding of glucocorticoid receptor (GR) homodimers. PMID- 11017169 TI - Specification of ventral neuron types is mediated by an antagonistic interaction between Shh and Gli3. AB - Specification of distinct neuron types in the ventral spinal cord is thought to be mediated by a graded concentration of Sonic hedgehog (Shh), a secreted signaling protein. Shh is made in the notochord, the most ventral part of the spinal cord, and in mice lacking Shh, ventral cell types are reduced or absent. The response to Shh depends on transcription factors of the Gli family, but the detailed mechanism is not understood. Here we show that Gli3 represses ventral fates in a dose-dependent manner. Whereas Shh -/- mutant mice show reductions in several classes of ventral interneurons and a complete absence of motor neurons, these cell types were rescued in Shh-/-;Gli3 -/- double mutants. This rescue of the Shh null phenotype depended on the level of Gli3 function; a partial rescue was observed in Shh-/-;Gli3 +/- embryos. We propose that Shh is required to antagonize Gli3, which would otherwise repress ventral fates. Differences between rostral and caudal regions suggest that other signaling molecules-in addition to Shh-may be involved in specifying ventral fates, particularly in the caudal region of the spinal cord. PMID- 11017170 TI - Skeletal myogenic potential of human and mouse neural stem cells. AB - Distinct cell lineages established early in development are usually maintained throughout adulthood. Thus, adult stem cells have been thought to generate differentiated cells specific to the tissue in which they reside. This view has been challenged; for example, neural stem cells can generate cells that normally originate from a different germ layer. Here we show that acutely isolated and clonally derived neural stem cells from mice and humans could produce skeletal myotubes in vitro and in vivo, the latter following transplantation into adult animals. Myogenic conversion in vitro required direct exposure to myoblasts, and was blocked if neural cells were clustered. Thus, a community effect between neural cells may override such myogenic induction. We conclude that neural stem cells, which generate neurons, glia and blood cells, can also produce skeletal muscle cells, and can undergo various patterns of differentiation depending on exposure to appropriate epigenetic signals in mature tissues. PMID- 11017171 TI - Concentration-dependent substate behavior of native AMPA receptors. AB - AMPA-type glutamate receptors mediate most excitatory postsynaptic currents (EPSCs) at central synapses, and their conductance determines in part the size of EPSCs. The conductance of a recombinant AMPA receptor depends on the number of agonist molecules bound to the channel. Here we tested whether native AMPA and kainate receptors show this behavior in outside-out patches from neurons in situ by measuring conductance levels of single channels over a wide range of agonist concentrations. We found that the conductance of AMPA, but not kainate, receptors depended strongly on agonist concentration. Our results suggest that alterations in the glutamate concentration in the synaptic cleft may change the apparent unitary conductance of postsynaptic AMPA receptors. PMID- 11017173 TI - Postsynaptic CPG15 promotes synaptic maturation and presynaptic axon arbor elaboration in vivo. AB - The formation of CNS circuits is characterized by the coordinated development of neuronal structure and synaptic function. The activity-regulated candidate plasticity gene 15 (cpg15) encodes a glycosylphosphatidylinositol (GPI)-linked protein whose in vivo expression increases the dendritic arbor growth rate of Xenopus optic tectal cells. We now demonstrate that tectal cell expression of CPG15 significantly increases the elaboration of presynaptic retinal axons by decreasing rates of branch retractions. Whole-cell recordings from optic tectal neurons indicate that CPG15 expression promotes retinotectal synapse maturation by recruiting functional AMPA receptors to synapses. Expression of truncated CPG15, lacking its GPI anchor, does not promote axon arbor growth and blocks synaptic maturation. These results suggest that CPG15 coordinately increases the growth of pre- and postsynaptic structures and the number and strength of their synaptic contacts. PMID- 11017172 TI - Transport rates of GABA transporters: regulation by the N-terminal domain and syntaxin 1A. AB - Plasma membrane GABA transporters participate in neural signaling through re uptake of neurotransmitter. The domains of the transporter that mediate GABA translocation and regulate transport are not well understood. In the present experiments, the N-terminal cytoplasmic domain of the GABA transporter GAT1 regulated substrate transport rates. This domain directly interacted with syntaxin 1A, a SNARE protein involved in both neurotransmitter release and modulation of calcium channels and cystic fibrosis transmembrane regulator (CFTR) chloride channels. The interaction resulted in a decrease in transporter transport rates. These data demonstrate that intracellular domains of the GABA and protein-protein interactions regulate substrate translocation, and identify a direct link between the machinery involved in transmitter release and re-uptake. PMID- 11017174 TI - Postsynaptic filopodia in muscle cells interact with innervating motoneuron axons. AB - Precise synaptogenesis is crucial to brain development, and depends on the ability of specific partner cells to locate and communicate with one another. Dynamic properties of axonal filopodia during synaptic targeting are well documented, but the cytomorphological dynamics of postsynaptic cells have received less attention. In Drosophila embryos, muscle cells bear numerous postsynaptic filopodia ('myopodia') during motoneuron targeting. Here we show that myopodia are actin-filled microprocesses, which progressively clustered at the site of motoneuron innervation while intermingling with presynaptic filopodia. In prospero mutants, which have severe delays in axon outgrowth from the CNS, myopodia were present initially but clustering behavior was not observed, demonstrating that clustering depends on innervating axons. Thus, postsynaptic filopodia are capable of intimate interaction with innervating presynaptic axons. We propose that, by contributing to direct long-distance cellular communication, they are dynamically involved in synaptic matchmaking. PMID- 11017175 TI - Enhancement of presynaptic neuronal excitability by correlated presynaptic and postsynaptic spiking. AB - Use-dependent modifications, such as long-term potentiation (LTP) of synaptic efficacy, are believed to be essential for information storage in the nervous system. Repetitive correlated spiking of presynaptic and postsynaptic neurons can induce LTP at excitatory glutamatergic synapses. In cultured hippocampal neurons, we show that repetitive correlated activity also results in a rapid and persistent enhancement of presynaptic excitability, decreasing the threshold for spiking and reducing the variability of interspike intervals. Furthermore, we found that correlated activity modified sodium channel gating in the presynaptic neuron. This modification of presynaptic excitability required a temporal order between presynaptic and postsynaptic spiking and activation of postsynaptic NMDA receptors. Presynaptic inhibition of protein kinase C abolished the change in excitability without affecting LTP. Such rapid activity-dependent changes in the efficacy of presynaptic spiking may be involved in the processing and storage of information within the nervous system. PMID- 11017176 TI - Cellular and network mechanisms of rhythmic recurrent activity in neocortex. AB - The neocortex generates periods of recurrent activity, such as the slow (0.1-0.5 Hz) oscillation during slow-wave sleep. Here we demonstrate that slices of ferret neocortex maintained in vitro generate this slow (< 1 Hz) rhythm when placed in a bathing medium that mimics the extracellular ionic composition in situ. This slow oscillation seems to be initiated in layer 5 as an excitatory interaction between pyramidal neurons and propagates through the neocortex. Our results demonstrate that the cerebral cortex generates an 'up' or depolarized state through recurrent excitation that is regulated by inhibitory networks, thereby allowing local cortical circuits to enter into temporarily activated and self-maintained excitatory states. The spontaneous generation and failure of this self-excited state may account for the generation of a subset of cortical rhythms during sleep. PMID- 11017177 TI - Odor maps in the mammalian olfactory bulb: domain organization and odorant structural features. AB - Psychophysical studies indicate that structural features of odorants differentially influence their perceived odor. In the olfactory bulb (OB), odorants are represented by ensembles of activated glomeruli. Here we used optical imaging of intrinsic signals to examine how these structural features are represented spatially in the sensory map of the rat OB. We found that the dorsal OB contained two topographically fixed domains; constituent glomeruli in each domain could be activated by odorants with particular functional groups. Within each domain, other structural features such as carbon chain length and branching were represented by local differences in patterns. These results suggest that structural features are categorized into two classes, primary features (functional groups) that characterize each domain, and secondary features that are represented by local positions within each domain. Such hierarchical representations of different structural features correlate well with psychophysical structure-odor relationships. PMID- 11017178 TI - Remote spatial memory in an amnesic person with extensive bilateral hippocampal lesions. AB - The hippocampus may have a time-limited role in memory, being needed only until information is permanently stored elsewhere, or this region may permanently represent long-term allocentric spatial information or cognitive maps in memory. To test these ideas, we investigated remote spatial memory in K.C., a patient with bilateral hippocampal lesions and amnesia for autobiographical events. In his spatial knowledge, general aspects were preserved, but details were lost, a pattern that resembled his memory loss in other domains. K.C. performed normally on allocentric spatial tests of his neighborhood and the world. He had difficulty, however, in recognizing and identifying non-salient neighborhood landmarks, and in recognizing city locations on world maps. This suggests that the hippocampus is not crucial for maintenance and retrieval of remotely formed spatial representations of major landmarks, routes, distances and directions, but is necessary for specifying location details, regardless of when they were acquired. PMID- 11017180 TI - RNA-protein machines rule. PMID- 11017179 TI - Subcortical and cortical brain activity during the feeling of self-generated emotions. AB - In a series of [15O]PET experiments aimed at investigating the neural basis of emotion and feeling, 41 normal subjects recalled and re-experienced personal life episodes marked by sadness, happiness, anger or fear. We tested the hypothesis that the process of feeling emotions requires the participation of brain regions, such as the somatosensory cortices and the upper brainstem nuclei, that are involved in the mapping and/or regulation of internal organism states. Such areas were indeed engaged, underscoring the close relationship between emotion and homeostasis. The findings also lend support to the idea that the subjective process of feeling emotions is partly grounded in dynamic neural maps, which represent several aspects of the organism's continuously changing internal state. PMID- 11017181 TI - (A)-tail of two polymerase structures. PMID- 11017182 TI - CooA, CAP and allostery. PMID- 11017183 TI - Mechanisms of amyloidogenesis. PMID- 11017184 TI - The road to RNase P. AB - In 1989, Sidney Altman and Thomas R. Cech shared the Nobel Prize in Chemistry for their discovery of catalytic properties of RNA. Cech was studying the splicing of RNA in a unicellular organism called Tetrahymena thermophila. He found that the precursor RNA could splice in vitro in the absence of proteins. Altman studied ribonuclease P (RNase P), a ribonucleoprotein that is a key enzyme in the biosynthesis of tRNA. RNase P is an RNA processing endonuclease that specifically cleaves precursors of tRNA, releasing 5' precursor sequences and mature tRNAs. RNase P is involved in processing all species of tRNA and is present in all cells and organelles that carry out tRNA synthesis. What follows is a personal recollection by Altman of how he came to study this remarkable enzyme. PMID- 11017185 TI - Picture story. A protein microarray. PMID- 11017186 TI - RNA: versatility in form and function. AB - RNA performs a remarkable range of functions in all cells. In addition to its central role in information transfer from DNA to protein, it is essential for functions as diverse as RNA processing, chromosome end-maintenance and dosage compensation. The versatility of RNA derives from its unique ability to use direct readout via base-pairing for sequence specific targeting (or templating) in combination with its capacity to form elaborate three dimensional structures. Such structures can perform catalysis or serve as protein recognition surfaces. In this short review, we attempt to give a flavor for the diversity of functional RNAs in the cell and highlight, using selected examples, two quite distinct activities, catalysis and sequence specific targeting. Within each section, we discuss how the lessons we have learned from these systems may apply to other, less well understood, RNAs. PMID- 11017187 TI - Induced fit in RNA-protein recognition. AB - Two generalizations can be drawn from the recent rapid progress in understanding RNA-protein interactions. First, there is a great diversity of observed protein and RNA structural motifs. Second, formation of almost every RNA-protein complex that has been characterized involves conformational changes in the protein, the RNA, or both. The role of these conformational changes in the biological function of RNA-protein complexes is not at all clear. Whether or not conformational changes are a critical feature of ribonucleoprotein complex assembly or are an unimportant mechanistic detail, the ubiquity of these changes warrants careful consideration of their implications. PMID- 11017188 TI - The ends of the affair: capping and polyadenylation. AB - Nearly all mRNAs are post-transcriptionally modified at their 5' and 3' ends, by capping and polyadenylation, respectively. These essential modifications are of course chemically quite distinct, as are the enzymatic complexes responsible for their synthesis. But recent studies have uncovered some similarities as well. For example, both involve entirely protein machinery, which is now the exception rather than the rule in RNA processing and modification reactions, and the two reactions share one important factor, namely RNA polymerase II. In this brief review, we describe progress in understanding the enzymes and factors that participate in these two processes, highlighting the evolutionary conservation, from yeast to humans, that has become apparent. PMID- 11017189 TI - Musing on the structural organization of the exosome complex. AB - The exosome complex of 3'-->5' exoribonucleases functions in both the precise processing of 3' extended precursor molecules to mature stable RNAs and the complete degradation of other RNAs. Both processing and degradative activities of the exosome depend on additional cofactors, notably the putative RNA helicases Mtr4p and Ski2p. It is not known how these factors regulate exosome function or how the exosome distinguishes RNAs destined for processing events from substrates that are to be completely degraded. Here we review the available data concerning the modes of action of the exosome and relate these to possible structural arrangements for the complex. As no detailed structural data are yet available for the exosome complex, or any of its constituent enzymes, this discussion will rely heavily on rather speculative models. PMID- 11017190 TI - The end of the (DNA) line. AB - Telomerases contain an essential RNA subunit (TER), as well as an essential protein reverse transcriptase subunit (TERT). The RNA subunit includes a short template region that is copied into telomeric DNA, but otherwise it is large and divergent. However, phylogenetic studies have revealed a conserved core secondary structure for TER. Much of the divergence can be accounted for by the acquisition of different types of RNA domains that function in RNA stabilization. Some of the nontemplate portions of TER, which include regions in the conserved core, are important for aspects of telomerase enzymatic activity independent of their role in telomerase assembly. Mutational studies indicate that telomerase enzyme function results from a collaboration of both protein and RNA functional groups contributed by TERT and TER. PMID- 11017191 TI - The question remains: is the spliceosome a ribozyme? AB - The two phosphoryl transfer steps of pre-mRNA splicing are catalyzed within the large ribonuclear protein machine called the spliceosome. The highly dynamic nature of the spliceosome has presented many challenges to a structural and mechanistic understanding of its catalytic core. While much evidence supports the popular hypothesis that the catalytic steps of pre-mRNA splicing are mediated by spliceosomal RNA, a role for protein in catalysis cannot yet be ruled out. A highly conserved protein, Prp8, is a component of the catalytic core. We review data consistent with the hypothesis that Prp8 functions as a cofactor to an RNA enzyme. PMID- 11017192 TI - Approaching translation at atomic resolution. AB - Atomic resolution structures of 50S and 30S ribosomal particles have recently been solved by X-ray diffraction. These ribosomal structures show often unusual folds of ribosomal RNAs and proteins, and provide molecular explanations for fundamental aspects of translation. In the 50S structure, the active site for peptide bond formation was localized and found to consist of RNA. The ribosome is thus a ribozyme. In the 30S structures, tRNA binding sites were located, and molecular mechanisms for ribosomal fidelity were proposed. The 30S subunit particle has three globular domains, and relative movements of these domains may be required for translocation of the ribosome during protein synthesis. The structures are consistent with and rationalize decades of biochemical analysis of translation and usher in a molecular age in understanding the ribosome. PMID- 11017193 TI - Functional defects of pathogenic human mitochondrial tRNAs related to structural fragility. AB - Aminoacylation of transfer RNAs (tRNAs) is essential for protein synthesis. A growing number of human diseases correlate with point mutations in tRNA genes within the mitochondrial genome. These tRNAs have unique sequences that suggest they have fragile structures. However, the structural significance of pathology related tRNA mutations and their effects on molecular function have not been explored. Here, opthalmoplegia related mutants of a human mitochondrial tRNA have been investigated. Each mutation replaces either an A-U or G-C pair in the predicted secondary structure with an A-C pair. Aminoacylation of each mutant tRNA was severely attenuated. Moreover, each strongly inhibited aminoacylation of the wild type substrate, suggesting that the effects of these mutations might not be bypassed in the potentially heteroplasmic environment of mitochondria. The function of mutant tRNAs was rescued by single compensatory mutations that restored Watson-Crick base pairing and reintroduced stability into regions of predicted secondary structure, even though the pairs introduced were different from those found in the wild type tRNA. Thus, functional defects caused by a subset of pathogenic mutations may result from the inherent structural fragility of human mitochondrial tRNAs. PMID- 11017194 TI - Functional sites of interaction between release factor RF1 and the ribosome. AB - Translational release factors decipher stop codons in mRNA and activate hydrolysis of peptidyl-tRNA in the ribosome during translation termination. The mechanisms of these fundamental processes are unknown. Here we have mapped the interaction of bacterial release factor RF1 with the ribosome by directed hydroxyl radical probing. These experiments identified conserved domains of RF1 that interact with the decoding site of the 30S ribosomal subunit and the peptidyl transferase site of the 50S ribosomal subunit. RF1 interacts with a binding pocket formed between the ribosomal subunits that is also the interaction surface of elongation factor EF-G and aminoacyl-tRNA bound to the A site. These results provide a basis for understanding the mechanism of stop codon recognition coupled to hydrolysis of peptidyl-tRNA, mediated by a protein release factor. PMID- 11017195 TI - Protein 4.1R core domain structure and insights into regulation of cytoskeletal organization. AB - The crystal structure of the core domain (N-terminal 30 kDa domain) of cytoskeletal protein 4.1R has been determined and shows a cloverleaf-like architecture. Each lobe of the cloverleaf contains a specific binding site for either band 3, glycophorin C/D or p55. At a central region of the molecule near where the three lobes are joined are two separate calmodulin (CaM) binding regions. One of these is composed primarily of an alpha-helix and is Ca 2+ insensitive; the other takes the form of an extended structure and its binding with CaM is dramatically enhanced by the presence of Ca 2+, resulting in the weakening of protein 4.1R binding to its target proteins. This novel architecture, in which the three lobes bind with three membrane associated proteins, and the location of calmodulin binding sites provide insight into how the protein 4.1R core domain interacts with membrane proteins and dynamically regulates cell shape in response to changes in intracellular Ca2+ levels. PMID- 11017196 TI - Structure of the CO sensing transcription activator CooA. AB - CooA is a homodimeric transcription factor that belongs to the catabolite activator protein (CAP) family. Binding of CO to the heme groups of CooA leads to the transcription of genes involved in CO oxidation in Rhodospirillum rubrum. The 2.6 A structure of reduced (Fe2+) CooA reveals that His 77 in both subunits provides one heme ligand while the N-terminal nitrogen of Pro 2 from the opposite subunit provides the other ligand. A structural comparison of CooA in the absence of effector and DNA (off state) with that of CAP in the effector and DNA bound state (on state) leads to a plausible model for the mechanism of allosteric control in this class of proteins as well as the CO dependent activation of CooA. PMID- 11017197 TI - von Willebrand factor conformation and adhesive function is modulated by an internalized water molecule. AB - Platelet participation in hemostasis and arterial thrombosis requires the binding of glycoprotein (GP) Ibalpha to von Willebrand factor (vWF). Hemodynamic forces enhance this interaction, an effect mimicked by the substitution I546V in the vWF A1 domain. A water molecule becomes internalized near the deleted Ile methyl group. The change in hydrophobicity of the local environment causes positional changes propagated over a distance of 27 A. As a consequence, a major reorientation of a peptide plane occurs in a surface loop involved in GP Ibalpha binding. This distinct vWF conformation shows increased platelet adhesion and provides a structural model for the initial regulation of thrombus formation. PMID- 11017198 TI - A switch in heme axial ligation prepares Paracoccus pantotrophus cytochrome cd1 for catalysis. AB - Cytochrome cd1 nitrite reductase (cd1) from Paracoccus pantotrophus is a respiratory enzyme capable of using nitrite, hydroxylamine and oxygen as electron accepting substrates. Structural studies have shown that when the enzyme is reduced there is a change in the axial ligation of both hemes, which has been proposed to form part of the catalytic cycle. Here we report the use of a physiological electron donor, pseudoazurin, to investigate the relationship between heme ligation and catalysis. A combination of visible absorption and electron paramagnetic resonance spectroscopies reveals the formation of a catalytically competent state of oxidized cd1 with 'switched' axial ligands immediately after complete reoxidation of reduced cd1 with hydroxylamine. This activated conformer returns over 20 min at 25 degrees C to the state previously observed for oxidized 'as isolated' cd1, which is catalytically inactive towards the same substrates. PMID- 11017199 TI - Structural basis for the diversity of DNA recognition by bZIP transcription factors. AB - The basic region leucine zipper (bZIP) proteins form one of the largest families of transcription factors in eukaryotic cells. Despite relatively high homology between the amino acid sequences of the bZIP motifs, these proteins recognize diverse DNA sequences. Here we report the 2.0 A resolution crystal structure of the bZIP motif of one such transcription factor, PAP1, a fission yeast AP-1-like transcription factor that binds DNA containing the novel consensus sequence TTACGTAA. The structure reveals how the Pap1-specific residues of the bZIP basic region recognize the target sequence and shows that the side chain of the invariant Asn in the bZIP motif adopts an alternative conformation in Pap1. This conformation, which is stabilized by a Pap1-specific residue and its associated water molecule, recognizes a different base in the target sequence from that in other bZIP subfamilies. PMID- 11017200 TI - Interactions within the yeast t-SNARE Sso1p that control SNARE complex assembly. AB - In the eukaryotic secretory and endocytic pathways, transport vesicles shuttle cargo among intracellular organelles and to and from the plasma membrane. Cargo delivery entails fusion of the transport vesicle with its target, a process thought to be mediated by membrane bridging SNARE protein complexes. Temporal and spatial control of intracellular trafficking depends in part on regulating the assembly of these complexes. In vitro, SNARE assembly is inhibited by the closed conformation adopted by the syntaxin family of SNAREs. To visualize this closed conformation directly, the X-ray crystal structure of a yeast syntaxin, Sso1p, has been determined and refined to 2.1 A resolution. Mutants designed to destabilize the closed conformation exhibit accelerated rates of SNARE assembly. Our results provide insight into the mechanism of SNARE assembly and its intramolecular and intermolecular regulation. PMID- 11017201 TI - Structural proteomics of an archaeon. AB - A set of 424 nonmembrane proteins from Methanobacterium thermoautotrophicum were cloned, expressed and purified for structural studies. Of these, approximately 20% were found to be suitable candidates for X-ray crystallographic or NMR spectroscopic analysis without further optimization of conditions, providing an estimate of the number of the most accessible structural targets in the proteome. A retrospective analysis of the experimental behavior of these proteins suggested some simple relations between sequence and solubility, implying that data bases of protein properties will be useful in optimizing high throughput strategies. Of the first 10 structures determined, several provided clues to biochemical functions that were not detectable from sequence analysis, and in many cases these putative functions could be readily confirmed by biochemical methods. This demonstrates that structural proteomics is feasible and can play a central role in functional genomics. PMID- 11017202 TI - The structure of the ubiquinol oxidase from Escherichia coli and its ubiquinone binding site. AB - Cell respiration is catalyzed by the heme-copper oxidase superfamily of enzymes, which comprises cytochrome c and ubiquinol oxidases. These membrane proteins utilize different electron donors through dissimilar access mechanisms. We report here the first structure of a ubiquinol oxidase, cytochrome bo3, from Escherichia coli. The overall structure of the enzyme is similar to those of cytochrome c oxidases; however, the membrane-spanning region of subunit I contains a cluster of polar residues exposed to the interior of the lipid bilayer that is not present in the cytochrome c oxidase. Mutagenesis studies on these residues strongly suggest that this region forms a quinone binding site. A sequence comparison of this region with known quinone binding sites in other membrane proteins shows remarkable similarities. In light of these findings we suggest specific roles for these polar residues in electron and proton transfer in ubiquinol oxidase. PMID- 11017204 TI - Technical reports PMID- 11017205 TI - Bioinformaticians develop new data mining tools PMID- 11017203 TI - Crystal structure of the class D beta-lactamase OXA-10. AB - We report the crystal structure of a class D beta-lactamase, the broad spectrum enzyme OXA-10 from Pseudomonas aeruginosa at 2.0 A resolution. There are significant differences between the overall fold observed in this structure and those of the evolutionarily related class A and class C beta-lactamases. Furthermore, the structure suggests the unique, cation mediated formation of a homodimer. Kinetic and hydrodynamic data shows that the dimer is a relevant species in solution and is the more active form of the enzyme. Comparison of the molecular details of the active sites of the class A and class C enzymes with the OXA-10 structure reveals that there is no counterpart in OXA-10 to the residues proposed to act as general bases in either of these enzymes (Glu 166 and Tyr 150, respectively). Our structures of the native and chloride inhibited forms of OXA 10 suggest that the class D enzymes have evolved a distinct catalytic mechanism for beta-lactam hydrolysis. Clinical variants of OXA-10 are also discussed in light of the structure. PMID- 11017206 TI - Effect of measurement on the decay rate of a quantum system AB - We investigated the electron tunneling out of a quantum dot in the presence of continuous monitoring by a detector. It is shown that the Schrodinger equation for the whole system can be reduced to new Bloch-type rate equations describing the time development of the detector and the measured system at once. Using these equations we find that the continuous measurement of the unstable system does not affect its exponential decay, exp(-gammat), contrary to expectations based on the quantum Zeno effect. However, the width of the energy distribution of the tunneling electron is no longer gamma, but increases due to the decoherence, generated by the detector. PMID- 11017207 TI - Wave packet dynamics in energy space, random matrix theory, and the quantum classical correspondence AB - We apply random-matrix-theory (RMT) to the analysis of evolution of wave packets in energy space. We study the crossover from ballistic behavior to saturation, the possibility of having an intermediate diffusive behavior, and the feasibility of strong localization effect. Both theoretical considerations and numerical results are presented. Using quantal-classical correspondence considerations we question the validity of the emerging dynamical picture. In particular, we claim that the appearance of the intermediate diffusive behavior is possibly an artifact of the RMT strategy. PMID- 11017208 TI - Observation of the scissors mode and evidence for superfluidity of a trapped bose einstein condensed Gas AB - We report the observation of the scissors mode of a Bose-Einstein condensed gas of 87Rb atoms in a magnetic trap, which gives direct evidence of superfluidity in this system. The scissors mode of oscillation is excited by a sudden rotation of the anisotropic trapping potential. For a gas above T(c) (normal fluid) we detect the occurrence of oscillations at two frequencies, with the lower frequency corresponding to the rigid body value of the moment of inertia. Well below T(c) the condensate oscillates at a single frequency, without damping, as expected for a superfluid. PMID- 11017209 TI - Density dependence of the transition temperature in a homogeneous bose-einstein condensate AB - Transition temperature data obtained as a function of particle density in the 4He Vycor system are compared with recent theoretical calculations for 3D Bose condensed systems. In the low density dilute Bose gas regime we find, in agreement with theory, a positive shift in the transition temperature of the form DeltaT/T0 = gamma(na(3))(1/3). At higher densities a maximum is found in the ratio of T(c)/T0 for a value of the interaction parameter, na(3), that is in agreement with path-integral Monte Carlo calculations. PMID- 11017210 TI - Is random close packing of spheres well defined? AB - Despite its long history, there are many fundamental issues concerning random packings of spheres that remain elusive, including a precise definition of random close packing (RCP). We argue that the current picture of RCP cannot be made mathematically precise and support this conclusion via a molecular dynamics study of hard spheres using the Lubachevsky-Stillinger compression algorithm. We suggest that this impasse can be broken by introducing the new concept of a maximally random jammed state, which can be made precise. PMID- 11017211 TI - Bayesian approach to inverse quantum statistics AB - A nonparametric Bayesian approach is developed to determine quantum potentials from empirical data for quantum systems at finite temperature. The approach combines the likelihood model of quantum mechanics with a priori information on potentials implemented in the form of stochastic processes. Its specific advantages are the possibilities to deal with heterogeneous data and to express a priori information explicitly in terms of the potential of interest. A numerical solution in maximum a posteriori approximation is obtained for one-dimensional problems. As nonparametric estimates, the results depend strongly on the implemented a priori information. PMID- 11017212 TI - Generalized second law in cosmology from causal boundary entropy AB - A classical and quantum mechanical generalized second law of thermodynamics in cosmology implies constraints on the effective equation of state of the universe in the form of energy conditions, obeyed by many known cosmological solutions, forbids certain cosmological singularities, and is compatible with entropy bounds. This second law is based on the conjecture that causal boundaries and not only event horizons have geometric entropies proportional to their area. In string cosmology the second law provides new information about nonsingular solutions. PMID- 11017213 TI - Phenomenology of a realistic accelerating universe using only planck-scale physics AB - Modern data are showing increasing evidence that the Universe is accelerating. So far, all attempts to account for the acceleration have required some fundamental dimensionless quantities to be extremely small. We show how a class of scalar field models (which may emerge naturally from superstring theory) can account for acceleration which starts in the present epoch with all the potential parameters O(1) in Planck units. PMID- 11017214 TI - Phenomenology of the randall-sundrum gauge hierarchy model AB - We explore the phenomenology of the localized gravity model of Randall and Sundrum where a 5-dimensional nonfactorizable geometry generates the gauge hierarchy by an exponential function called a warp factor. The Kaluza-Klein (KK) tower of gravitons in this scenario has different properties from those in factorizable models. We derive the KK graviton interactions with the standard model fields and obtain constraints from their direct production at hadron colliders as well as from virtual KK exchanges. We study the KK spectrum in e(+)e(-) annihilation and show how to determine the model parameters if the first KK state is observed. PMID- 11017215 TI - Holography and noncommutative yang-mills theory AB - In this Letter a recently proposed gravity dual of noncommutative Yang-Mills theory is derived from the relations between closed string moduli and open string moduli recently suggested by Seiberg and Witten. The only new input one needs is a simple form of the running string tension as a function of energy. This derivation provides convincing evidence that string theory integrates with the holographical principle and demonstrates a direct link between noncommutative Yang-Mills theory and holography. PMID- 11017216 TI - Search for second-generation leptoquark pairs in &pmacr;p collisions at radicals = 1.8 TeV AB - We have searched for second-generation leptoquark (LQ) pairs in the &mgr;&mgr;+jets channel using 94+/-5 pb(-1) of &pmacr;p collider data collected by the D0 experiment at the Fermilab Tevatron during 1993-1996. No evidence for a signal is observed. These results are combined with those from the &mgr;nu+jets and nunu+jets channels to obtain 95% confidence level (C.L.) upper limits on the LQ pair production cross section as a function of mass and beta, the branching fraction of a LQ decay into a charged lepton and a quark. Lower limits of 200(180) GeV/c(2) for beta = 1(1 / 2) are set at the 95% C.L. on the mass of scalar LQ. Mass limits are also set on vector leptoquarks as a function of beta. PMID- 11017217 TI - Production of Upsilon(1S) mesons from chi(b) decays in p&pmacr; collisions at sqrt AB - We have reconstructed the radiative decays chi(b)(1P)-->Upsilon(1S)gamma and chi(b)(2P)-->Upsilon(1S)gamma in p&pmacr; collisions at sqrt[s] = 1.8 TeV, and measured the fraction of Upsilon(1S) mesons that originate from these decays. For Upsilon(1S) mesons with p(Upsilon)(T)>8.0 GeV/c, the fractions that come from chi(b)(1P) and chi(b)(2P) decays are [27.1+/-6.9(stat)+/-4. 4(syst)]% and [10.5+/ 4.4(stat)+/-1.4(syst)]%, respectively. We have derived the fraction of directly produced Upsilon(1S) mesons to be [50.9+/-8.2(stat)+/-9.0(syst)]%. PMID- 11017218 TI - Hard-thermal-loop resummed pressure of a degenerate quark-gluon plasma AB - We compute the pressure of a finite-density quark-gluon plasma at zero temperature to leading order in hard-thermal-loop perturbation theory, which includes the fermionic excitations and Landau damping. The result is compared with the weak-coupling expansion for finite positive chemical potential &mgr; through order alpha(2)(s) and with a quasiparticle model with a mass depending on &mgr;. PMID- 11017219 TI - Half-life measurement for the rp-process waiting point nuclide 80Zr AB - X-ray bursts have been suggested as a possible site for the astrophysical rp process. The time scale for the process is governed by beta-decay half-lives of several even-even N = Z waiting point nuclei, in particular, N = Z = 40 80Zr. A 4.1(+0.8/-0.6)-s beta(+)/EC half-life for 80Zr was determined by observing delayed 84-keV gamma rays depopulating a T(1/2) = 4-&mgr;s isomer at 312 keV in the daughter 80Y. As this half-life is lower than many previously predicted values, the calculated excessive production of A = 80 nuclides in astrophysical x ray burst scenarios is reduced, and less extreme conditions are necessary for the production of heavier nuclides. PMID- 11017220 TI - Can the eigenstates of a many-body hamiltonian Be represented exactly using a general two-body cluster expansion? AB - Using a general two-body exponential parametrization for the wave function, the Nakatsuji two-particle density equation [Phys. Rev. A 14, 41 (1976)] is transformed into a set of nonlinear algebraic equations in which the number of unknowns precisely equals the number of equations. Since the Nakatsuji two particle density equation is equivalent to the time-independent Schrodinger equation for Hamiltonians containing up to two-body interactions, the answer to the title question is affirmative, provided the equations have solutions. Practical implications of the parametrization and possible approximation schemes are briefly discussed. PMID- 11017221 TI - Strong-coupling theory of two-level atoms in periodic fields AB - We present a new convergent strong-coupling expansion for two-level atoms in external periodic fields, free of secular terms. As a first application, we show that the coherent destruction of tunneling is a third-order effect. We also present an exact treatment of the high-frequency region, and compare it with the theory of averaging. The qualitative frequency spectrum of the transition probability amplitude contains an effective Rabi frequency. PMID- 11017222 TI - Electric-octupole and pure-electric-quadrupole effects in soft-X-Ray photoemission AB - Second-order [ O(k(2)), k = omega/c] nondipole effects in soft-x-ray photoemission are demonstrated via an experimental and a theoretical study of angular distributions of neon valence photoelectrons in the 100-1200 eV photon energy range. A newly derived theoretical expression for nondipolar angular distributions characterizes the second-order effects using four new parameters with primary contributions from pure-quadrupole and octupole-dipole interference terms. Independent-particle calculations of these parameters account for a significant portion of the existing discrepancy between experiment and theory for Ne 2p first-order nondipole parameters. PMID- 11017223 TI - Unique case of highly polarized collision-induced light scattering: the very far spectral wing by the helium pair AB - Contrary to what has been observed thus far collision-induced light scattering (CILS) can be completely polarized. This exceptional behavior characterizes the very far wing of the binary CILS spectrum by gaseous helium. This conclusion is drawn from an experimental study of the depolarization ratio of He (2) in a much extended, previously unexplored, spectral domain. Our analysis shows that this property, unique thus far, is mainly due to an almost perfect cancellation between polarization and exchange pair polarizability contributions to the depolarized spectrum, taking place at internuclear distances shorter than the atomic diameter. PMID- 11017224 TI - Unexpected behavior of the stopping of slow ions in ionic crystals AB - The energy loss of slow ions during grazing scattering from a LiF(100) surface as a function of the projectile atomic number Z1 is observed to show oscillations similar to those occurring in metals. A model of stopping of ions in an electron gas where screening is calculated from density functional theory reproduces well the experimental data. The same model gives good agreement with the energy loss obtained in transmission experiments performed with H and He projectiles. Analysis of these results allows us to gain new insights in the stopping of slow ions in ionic crystals. PMID- 11017225 TI - From above threshold ionization to statistical electron emission: the laser pulse duration dependence of C60 photoelectron spectra AB - The photoelectron spectra of C60 ionized using a 790 nm laser with pulse durations varying from 25 fs to 5 ps have been determined. For 25 fs pulses, in the absence of fragmentation, the ionization mechanism is direct multiphoton ionization with clear observation of above threshold ionization. As the pulse duration is increased, this becomes dominated by a statistical ionization due to equilibration among the electronic degrees of freedom. For pulse durations on the order of a ps coupling to the vibrational degrees of freedom occurs and the well known phenomenon of delayed (&mgr;s) ionization is observed. PMID- 11017226 TI - Local estimates for entropy densities in coupled map lattices AB - We present a method to derive an upper bound for the entropy density of coupled map lattices with local interactions from local observations. To do this, we use an embedding technique that is a combination of time delay and spatial embedding. This embedding allows us to identify the local character of the equations of motion. Based on this method we present an approximate estimate of the entropy density by the correlation integral. PMID- 11017227 TI - Spontaneous radiation and lamb shift in three-dimensional photonic crystals AB - Spontaneous emission in photonic crystals with anisotropic three-dimensional dispersion relation is studied. If the upper level is below a characteristic frequency omega(1), or above omega(2), or between omega(1) and omega(2), the radiation is a localized field with a frequency in the band gap, or a propagating field with a frquency in the band, or a diffusion field, respectively. An analytical expression for the Lamb shift is obtained. The Lamb shift for the current case is small compared to that in an ordinary vacuum or in one- or two dimensional photonic crystals due to lower density of states. PMID- 11017228 TI - Tight-binding description of the coupled defect modes in three-dimensional photonic crystals AB - We have experimentally observed the eigenmode splitting due to coupling of the evanescent defect modes in three-dimensional photonic crystals. The splitting was well explained with a theory based on the classical wave analog of the tight binding (TB) formalism in solid state physics. The experimental results were used to extract the TB parameters. A new type of waveguiding in a photonic crystal was demonstrated experimentally. A complete transmission was achieved throughout the entire waveguiding band. We have also obtained the dispersion relation for the waveguiding band of the coupled periodic defects from the transmission-phase measurements and from the TB calculations. PMID- 11017229 TI - Finite thermal conductivity in 1D lattices AB - We discuss the thermal conductivity of a chain of coupled rotators, showing that it is the first example of a 1D nonlinear lattice exhibiting normal transport properties in the absence of an on-site potential. Numerical estimates obtained by simulating a chain in contact with two thermal baths at different temperatures are found to be consistent with those based on linear response theory. The dynamics of the Fourier modes provides direct evidence of energy diffusion. The finiteness of the conductivity is traced back to the occurrence of phase jumps. Our conclusions are confirmed by the analysis of two variants of this model. PMID- 11017230 TI - Lattice-boltzmann model with sub-grid-scale boundary conditions AB - A lattice-Boltzmann method has been developed to incorporate solid-fluid boundary conditions on length scales less than the grid spacing. By introducing a continuous parameter, specified at each node and representing the fluid volume fraction associated with that node, we obtain second-order accuracy for boundaries at arbitrary positions and orientations with respect to the grid. The method does not require surface normals, and can therefore be applied to irregular geometries such as porous media. The new rules conserve mass and momentum, and reduce to the link bounce-back rule at aligned interfaces. PMID- 11017231 TI - Small-action particles in a tokamak in the presence of an n = 1 mode AB - It is found that an m = n = 1 mode with amplitude exceeding a certain threshold can lead to stochastic motion of energetic ions in tokamaks, the large orbit width particles (potatoes) being most easily affected. An n = 1 mode can redistribute particles also in the absence of stochasticity but only when the perturbation is quickly switched on/off, e.g., during a sawtooth crash. In the latter case, the perturbation results in a regular motion of particles around a certain helical orbit, at which a resonance driven by the mode but having no amplitude threshold takes place. PMID- 11017232 TI - Hydrogen adatoms on TiO2(110)-(1x1) characterized by scanning tunneling microscopy and electron stimulated desorption AB - Hydrogen atoms adsorbed on TiO2(110)-(1x1) surfaces have been characterized by scanning tunneling microscopy (STM) combined with electron stimulated desorption (ESD) technique. Certain amounts of H atoms are unexpectedly found on the TiO2 surfaces annealed at 900 K. Two forms of adsorption were discriminated in STM images from the different sensitivity to ESD and tentatively assigned to hydroxyl type (O-H) and hydride-type (Ti-H) species. PMID- 11017233 TI - Extension of rod-coil multiblock copolymers and the effect of the helix-coil transition AB - The extension elasticity of rod-coil mutliblock copolymers is analyzed for two experimentally accessible situations. In the quenched case, when the architecture is fixed by the synthesis, the force law is distinguished by a sharp change in the slope. In the annealed case, where interconversion between rod and coil states is possible, the resulting force law is sigmoid with a pronounced plateau. This last case is realized, for example, when homopolypeptides capable of undergoing a helix-coil transition are extended from a coil state. PMID- 11017234 TI - Shear induced smectic- A-smectic- C transition in side-chain liquid-crystalline polymers AB - We show that the smectic layer orientation in side-chain liquid-crystalline polymers depends on the molecular weight and the shear rate. Parallel orientation is obtained with the low molecular weight polymer and reveals a shear induced decrease of the smectic layer thickness. In contrast with usual liquid crystals, the shear torque acts at the mesogen scale through an efficient coupling between the polymer main chain and the mesogens resulting in a smectic- A-smectic- C transition. PMID- 11017235 TI - Metastable states as a key to the dynamics of supercooled liquids AB - Computer simulations of a model glass-forming system are presented, which study the correlation between the dynamics in real space and the topography of the potential energy landscape. This analysis clearly reveals that in the supercooled regime the dynamics is strongly influenced by the presence of deep valleys in the energy landscape, corresponding to long-lived metastable amorphous states. We explicitly relate nonexponential relaxation effects and dynamic heterogeneities to these metastable states and thus to the specific topography of the energy landscape. PMID- 11017236 TI - Vacancy interaction with dislocations in silicon: the shuffle-glide competition AB - Competition between the two alternative positions (shuffle and glide 111 plane subsets) for the core of a 30 degrees partial dislocation in Si is examined. Using a combination of ab initio total energy calculations with finite temperature free-energy calculations based on an interatomic potential, we obtained free energies for the relevant vacancy-type core defects. Generally, the free energy of vacancy formation in the core of a 30 degrees glide partial dislocation is considerably lower (by more than 1 eV) than in the bulk. However, even at high temperatures, the predicted thermal concentration of the shuffle segments comprised of a row of vacancies in the core is low, placing the 30 degrees partial dislocation in the glide subset position. PMID- 11017237 TI - Anomalous frequency dependence of the internal friction of vitreous silica AB - The internal friction Q-1 and the sound velocity deltav/v of vitreous silica were measured at very low temperatures using mechanical double paddle resonators operated at frequencies ranging from 0.33 to 14 kHz. Below approximately 40 mK the internal friction showed an unexpected temperature and frequency dependence, with absolute values of Q-1 clearly exceeding those predicted by the standard tunneling model. Even though the most plausible origin of the observed excess internal friction appears to be the mutual interaction between tunneling states, the results are difficult to reconcile quantitatively with present theories taking into account this interaction. PMID- 11017238 TI - Critical adsorption on defects in ising magnets and binary alloys AB - Long-range correlations in a magnet close to its critical point or in a binary alloy close to a continuous order-disorder transition can substantially enhance the effect of local perturbations. It is demonstrated using a position-space renormalization procedure that quasi-one-dimensional defects which break the symmetry of the order parameter have pronounced effects: They cause long-range critical adsorption profiles and give rise to new universal critical exponents, which are identified and calculated using field-theoretical methods. PMID- 11017239 TI - Surfactant effects and an order-disorder transition in binary gas-liquid nucleation AB - Density-functional theory is applied to the interaction site model to study gas liquid nucleation in a partially immiscible binary system consisting of spherical Lennard-Jones atoms (monomers) and dumbbell molecules of two Lennard-Jones atoms (dimers). Increased interaction anisotropy between the dimer sites and monomers is shown to result in mutual enhancement of nucleation. Critical nuclei with a lamellar structure are observed at high dimer activities. PMID- 11017240 TI - Scaling of the conductivity spectra in ionic glasses: dependence on the structure AB - A scaling approach in the conductivity formalism is applied to lithium tellurite glasses of different compositions. We observe that the hopping frequency can be used as the scaling frequency in the absence of well-defined dielectric loss peaks, and a universality of the scaling of the conductivity can be obtained for compositions with a similar structure. Further, the reasons behind the inapplicability of the scaling approach reported recently are elucidated in terms of structure of the glasses. PMID- 11017241 TI - Subcritical dissipation in three-dimensional superflows AB - Three-dimensional (3D) superflows past a circular cylinder are studied by numerically integrating the nonlinear Schrodinger equation. 3D initial data are built from the two-dimensional (2D) stationary vortex nucleation solutions. Quasistationary half-ring vortices, pinned at the sides of the cylinder, are generated after a short time. On a longer time scale, either 3D vortex stretching induces dissipation and drag, or the vortex is absorbed by the cylinder. The corresponding 3D critical velocity is found to be well below the 2D one. The implications for experiments in Bose-Einstein condensed gas and low-temperature helium are discussed. PMID- 11017242 TI - Enhanced heat capacity and a new temperature instability in superfluid 4He in the presence of a constant heat flux near t(lambda) AB - We present the first experimental evidence that the heat capacity of superfluid 4He, at temperatures very close to the lambda point T(lambda), is enhanced by a constant heat flux Q. The heat capacity at constant Q, C(Q), is predicted to diverge at a temperature T(c)(Q)B(p). A semiclassical drift-diffusion model gives a good description of the effects of the magnetic confinement on both the diagonal and off-diagonal components of the resistivity tensor. PMID- 11017252 TI - Role of the anisotropy of lead in BSCCO-Pb Josephson tunneling. AB - Recent experimental studies of Josephson tunneling between single crystals of Bi2Sr2CaCu2O8+x (BSCCO) and lead (Pb) films have revealed small, but finite, critical currents along the c axis of BSCCO, despite this being forbidden by symmetry. We show that the known anisotropy of the Pb order parameter would allow quite strong coupling between single crystals of BSCCO and Pb if the tunneling direction were along the BSCCO c axis and the [110] direction of the Pb crystal. This mechanism could account for the experimental results on granular Pb films if there is a few percent preferred orientation in the films. All the current experimental evidence is therefore consistent with BSCCO being a pure d-wave superconductor. PMID- 11017253 TI - Direct magneto-optical observation of a structural phase transition in thin films of manganites. AB - The spontaneous formation of twins in La(2/3)Ca(1/3)MnO3 films below T(S) approximately 105 K is observed by a magneto-optical technique. The twins are revealed as stripes along the {110} directions where magnetization tilts out of the film plane due to the stresses in twins. Their appearance is associated with a martensitic phase transition in the film triggered by the cubic-to-tetragonal transition in the SrTiO3 substrate. It is found that magnetization of the films proceeds by inhomogeneous rotation of magnetic moments. This is due to the presence of microscopic structural inhomogeneities. Their dominating role in the low-temperature transport can explain small effects of the transition at T(S) on the resistivity. PMID- 11017254 TI - Reduction of magnetic moments in very thin Cr layers of Fe/Cr multilayers: evidence from 119Sn Mossbauer spectroscopy. AB - Fe/Cr multilayers with monatomic Sn layers embedded in the Cr layers were grown epitaxially on MgO(001) substrates, and the magnetic hyperfine field at the 119Sn nuclear sites was examined using Mossbauer spectroscopy. It was found that nonzero hyperfine field is induced at the Sn sites at room temperature and that the value reduces drastically from 10 to 2 T when the Cr layer thickness decreases from 80 to 10 A. The result indicates that the Cr layers are magnetically ordered even when the thickness is very small and that the magnetic moments of Cr become smaller as the Cr layer thickness decreases. PMID- 11017255 TI - Two-step disordering of perpendicularly magnetized ultrathin films. AB - We have imaged the stripe domain structure of perpendicularly magnetized fcc ultrathin Fe films grown on Cu(100). The stripe phase has a strong local orientational order and sustains the two kinds of fluctuations predicted by Abanov et al. [Phys. Rev. B 51, 1023 (1995)]: meandering and dislocations. Before reaching the Curie temperature, the stripes transform into a new and so far unobserved domain structure, characterized by domains with predominantly square corners. We argue that this phase is the tetragonal liquid phase proposed by Abanov et al. to separate the stripe phase from the paramagnetic phase. This two step disordering is reminiscent of a two-dimensional melting process. PMID- 11017256 TI - Novel behavior of bond-centered muonium in heavily doped n-type silicon: Curie like spin susceptibility and charge screening. AB - Bond-centered muonium ( Mu(0)(BC)) has been observed in very heavily doped n-type Si:P. It exhibits a Curie-like electronic spin susceptibility which leads to a giant negative shift in the muon spin precession frequency. At high dopant levels, the Mu(0)(BC) hyperfine parameters, deduced from a model involving spin exchange with free carriers, are significantly reduced from those in intrinsic Si. This indicates that the spin density distribution for Mu(0)(BC) in metallic Si:P is altered significantly by charge screening effects, likely a general phenomenon for deep impurities in materials with high carrier concentrations. PMID- 11017257 TI - Selective photon-stimulated desorption of hydrogen from GaAs surfaces. AB - Photon-stimulated desorption of H(+) from hydrogenated GaAs (110) and (100) surfaces was studied as a function of photon energy. Distinct peaks, observed around As 3d core-level binding energy for desorption from the GaAs (100) surface and in the As 3d and Ga 3p region for desorption from the GaAs (110) surface, show a striking similarity with the fine structure (spin-orbit splitting) measured in the photoemission from As 3d and Ga 3p levels. These results provide clear evidence for direct desorption processes and represent a basis for selective modification of hydrogenated GaAs surfaces. PMID- 11017258 TI - Atomic many-body effects for the p-shell photoelectron spectra of transition metals. AB - Ab initio theoretical results for the 2p- and 3p-hole states of an Mn(2+) ion are reported in order to determine the importance of atomic contributions to the photoelectron spectra of bulk MnO. A combined treatment of relativity and electron correlation reveals important physical effects that have been neglected in virtually all previous work. The many-body and relativistic effects included in the atomic model are able, without any ad hoc empirical parameters, to explain most of the features of the MnO photoelectron spectra. In particular, it is not necessary to invoke charge transfer to explain the complex p-level spectra. PMID- 11017259 TI - Information, relative entropy of entanglement, and irreversibility. AB - Previously proposed measures of entanglement, such as entanglement of formation and assistance, are shown to be special cases of the relative entropy of entanglement. The difference between these measures for an ensemble of mixed states is shown to depend on the availability of classical information about particular members of the ensemble. Based on this, relations between relative entropy of entanglement and mutual information are derived. PMID- 11017260 TI - dc = 4 is the upper critical dimension for the Bak-Sneppen model. AB - Numerical results are presented indicating d(c) = 4 as the upper critical dimension for the Bak-Sneppen evolution model. This finding agrees with previous theoretical arguments, but contradicts a recent Letter [Phys. Rev. Lett. 80, 5746 (1998)] that placed d(c) as high as d = 8. In particular, we find that avalanches are compact for all dimensions d< or =4 and are fractal for d>4. Under those conditions, scaling arguments predict a d(c) = 4, where hyperscaling relations hold for d< or =4. Other properties of avalanches, studied for 1< or =d< or =6, corroborate this result. To this end, an improved numerical algorithm is presented that is based on the equivalent branching process. PMID- 11017261 TI - Pumped biochemical reactions, nonequilibrium circulation, and stochastic resonance. AB - Based on a master equation formalism for mesoscopic, unimolecular biochemical reactions, we show the periodic oscillation arising from severe nonequilibrium pumping is intimately related to the periodic motion in recently studied stochastic resonance (SR). The white noise in SR is naturally identified with the temperature in the biochemical reactions; the drift in the SR is associated with the circular flux in nonequilibrium steady state (NESS). As in SR, an optimal temperature for biochemical oscillation is shown to exist. A unifying framework for Hill's theory of NESS and the SR without periodic forcing is presented. The new formalism provides an analytically solvable model for SR. PMID- 11017262 TI - Universal aging features in the restructuring of fractal colloidal gels. AB - We use multispeckle dynamic light scattering to measure the dynamic structure factor, f(q,tau), of gels formed by aggregation of colloids. Although the gel is an elastic solid, f(q,tau) nearly completely decays on long time scales, with an unusual form, f(q, tau) approximately exp{-(tau/tau(f))(mu)}, with mu approximately 1.5 and with tau(f) proportional variant q(-1). A model for restructuring of the gel with aging correctly accounts for this behavior. Aging leads to a dramatic increase in tau(f); however, all data can be scaled on a single master curve, with tau(f) asymptotically growing linearly with age. This behavior is strikingly similar to that predicted for aging in disordered glassy systems, offering convincing proof of the universality of these concepts. PMID- 11017263 TI - Resistive anomalies at ferromagnetic transitions revisited: the case of SrRuO3. PMID- 11017265 TI - Comment on "Quantum theory of secondary emission in optically excited semiconductor quantum wells". PMID- 11017267 TI - Bose-einstein condensation in quasi-2D trapped gases AB - We discuss Bose-Einstein condensation (BEC) in quasi-2D trapped gases and find that well below the transition temperature T(c) the equilibrium state is a true condensate, whereas at intermediate temperatures T2pi amplitudes, using the measured pi-pi phase shifts with J = 0 and I = 0,2. The final rescattering of the two pions provides a strong enhancement of the DeltaI = 1/2 amplitude, which so far has been neglected in the theoretical predictions of varepsilon(')/varepsilon. This correction increases the standard model prediction of varepsilon(')/varepsilon to values in good agreement with the experimental measurements. PMID- 11017272 TI - Neutrino mass anarchy AB - What is the form of the neutrino mass matrix which governs the oscillations of the atmospheric and solar neutrinos? Features of the data have led to a dominant viewpoint where the mass matrix has an ordered, regulated pattern, perhaps dictated by a flavor symmetry. We challenge this viewpoint and demonstrate that the data are well accounted for by a neutrino mass matrix which appears to have random entries. PMID- 11017273 TI - Search for free fractional electric charge elementary particles using an automated millikan oil drop technique AB - We have carried out a direct search in bulk matter for free fractional electric charge elementary particles using the largest mass single sample ever studied about 17.4 mg of silicone oil. The search used an improved and highly automated Millikan oil drop technique. No evidence for fractional charge particles was found. The concentration of particles with fractional charge more than 0. 16e ( e being the magnitude of the electron charge) from the nearest integer charge is less than 4.71x10(-22) particles per nucleon with 95% confidence. PMID- 11017274 TI - New measurement of the rare decay K+ --> pi(+)&mgr;(+)&mgr;(-) AB - More than 400 K+-->pi(+)&mgr;(+)&mgr;(-) events were observed in a rare K+ decay experiment at the Brookhaven Alternating Gradient Synchrotron. Normalized to the K+-->pi(+)pi(+)pi(-) decay, the branching ratio is determined to be [9.22+/ 0.60(stat)+/-0. 49(syst)]x10(-8). This branching ratio and the &mgr;&mgr; mass spectrum is in very good agreement with the measurement of the K+-->pi(+)e(+)e(-) decay, but deviates significantly from the previous measurement. PMID- 11017275 TI - Measurement of the spin asymmetry in the photoproduction of pairs of high- p(T) hadrons at HERMES AB - We present a measurement of the longitudinal spin asymmetry A(||) in photoproduction of pairs of hadrons with high transverse momentum p(T). Data were accumulated by the HERMES experiment using a 27.5 GeV polarized positron beam and a polarized hydrogen target internal to the HERA storage ring. For h(+)h(-) pairs with p(h(1))(T)>1.5 GeV/c and p(h(2))(T)>1.0 GeV/c, the measured asymmetry is A(||) = -0. 28+/-0.12(stat)+/-0.02(syst). This negative value is in contrast to the positive asymmetries typically measured in deep inelastic scattering from protons, and is interpreted to arise from a positive gluon polarization. PMID- 11017276 TI - Hard exclusive electroproduction of decuplet baryons in the large N(c) limit AB - The cross sections and transverse spin asymmetries in the hard exclusive electroproduction of decuplet baryons are calculated in the large N(c) limit and found to be comparable to that of octet baryons. Large N(c) selection rules for the production amplitudes are derived, leading to new sensitive tests of the spin aspects of the QCD chiral dynamics both in the nonstrange and strange sectors. Importance of such studies for the reliable extraction of the pion form factor from pion electroproduction is explained. PMID- 11017277 TI - Search for the weak decay of a lightly bound H0 dibaryon AB - We present results of a search for a new form of hadronic matter, a six-quark, dibaryon state called the H0, a state predicted to exist in several theoretical models. Analyzing data collected by experiment E799-II at Fermilab, we searched for the decay H0-->Lambdappi(-) and found no candidate events. We place an upper limit on [B(H0-->Lambdappi(-))dsigma(H)/dOmega]/(dsigma(Xi)/dOmega) and, in the context of published models, exclude the region of lightly bound mass states just below the LambdaLambda mass threshold, 2.194hadrons)/sigma(e(+)e(-)-->&mgr;(+)&mgr;(-)), are determined. PMID- 11017325 TI - Gapless color superconductivity AB - We present the dispersion relations for quasiparticle excitations about the color flavor locked ground state of QCD at high baryon density. In the presence of condensates which pair light and strange quarks there need not be an energy gap in the quasiparticle spectrum. This raises the possibility of gapless color superconductivity, with a Meissner effect but no minimum excitation energy. Analysis within a toy model suggests that gapless color superconductivity may occur only as a metastable phase. PMID- 11017326 TI - Thermodynamics of a n-p condensate in asymmetric nuclear matter AB - We study the neutron-proton pairing in nuclear matter as a function of isospin asymmetry at finite temperatures and the empirical saturation density using realistic nuclear forces and Brueckner-renormalized single particle spectra. Our computation of the thermodynamic quantities shows that, while the difference of the entropies of the superconducting and normal phases anomalously changes its sign as a function of temperature for arbitrary asymmetry, the grand canonical potential does not; the superconducting state is found to be stable in the whole temperature-asymmetry plane. The pairing gap completely disappears for density asymmetries exceeding alpha(c) = (rho(n)-rho(p))/rho approximately 0.11. PMID- 11017327 TI - Three-nucleon force and the A(y) puzzle in intermediate energy p--> + d and d--> + p elastic scattering AB - New vector analyzing-power data on p-->+d elastic scattering at E(p) = 150 and 190 MeV have been measured. These are presented together with existing data and with recent d-->+p vector and tensor analyzing power data at E(d) = 270 MeV. The strong negative extremum of both vector analyzing powers A(p)(y) and A(d)(y) at straight theta(c.m.) approximately 80 degrees -120 degrees is underestimated by Faddeev calculations using modern NN forces. Inclusion of the Tucson-Melbourne 3N force shifts the minima upwards, but with conflicting results for A(p)(y), and leading to a good description for A(d)(y). An A(p)(y) puzzle, previously thought to exist at energies E(N)3) are derived from T2 and T3. This closure scheme is realized by a non-Gaussian statistical model. We use it to study the scaling law of T(p), and we find that the available data on scaling exponents favor Kolmogorov's 1941 theory rather than his 1962 theory. We also predict the high-order universal constants of inertial-range scaling. PMID- 11017338 TI - Holtsmark distributions in point-vortex systems AB - The statistics of uncorrelated point vortices in a plane is studied analytically and numerically. Theoretical distributions are obtained with the general method developed by Holtsmark [Ann. Phys. 58, 577 (1919)] and Chandrasekhar [Rev. Mod. Phys. 15, 1 (1943)]. They are found to agree with the results of numerical tests. Randomly placed Euler vortices have nearly Gaussian velocity distributions and Lorentzian distributions of the velocity difference. Statistics of other types of point vortices is essentially non-Gaussian. PMID- 11017339 TI - Two-mode rhomboidal states in driven surface waves AB - Two-mode rhomboid patterns are generated experimentally via two-frequency parametric forcing of surface waves. These patterns are formed by the simple nonlinear resonance: k-->'2-k-->(2) = k-->(1) where k(1) and k(2)( = k(')(2)) are concurrently excited eigenmodes. The state possesses a direction-dependent time dependence described by a superposition of the two modes, and a dimensionless scaling delineating the state's region of existence is presented. We also show that 2n-fold quasipatterns naturally arise from these states when the coupling angle between k-->(2) and k-->'2 is 2pi/n. PMID- 11017340 TI - Emittance growth during bunch compression in the CTF-II AB - Measurements of the beam emittance during bunch compression in the CLIC Test Facility (CTF-II) are described. The measurements were made with different beam charges and different energy correlations versus the bunch compressor settings which were varied from no compression through the point of full compression and to overcompression. Significant increases in the beam emittance were observed with the maximum emittance occurring near the point of full (maximal) compression. Finally, evaluation of possible emittance dilution mechanisms indicates that coherent synchrotron radiation was the most likely cause. PMID- 11017341 TI - Sustained kilowatt lasing in a free-electron laser with same-cell energy recovery AB - Jefferson Laboratory's kW-level infrared free-electron laser utilizes a superconducting accelerator that recovers about 75% of the electron-beam power. In achieving first lasing, the accelerator operated "straight ahead" to deliver 38-MeV, 1.1-mA cw current for lasing near 5 &mgr;m. The waste beam was sent directly to a dump while producing stable operation at up to 311 W. Utilizing the recirculation loop to send the electron beam back to the linac for energy recovery, the machine has now recovered cw average currents up to 5 mA, and has lased cw with up to 1720 W output at 3.1 &mgr;m. PMID- 11017342 TI - First laboratory observation of freely propagating electrostatic waves produced by intense, localized field structures AB - Freely propagating electrostatic waves (free modes) are observed to be produced by intense, localized field structures in laboratory measurements performed in an unmagnetized plasma in a state of strong Langmuir turbulence. The freely traveling waves conform to the Langmuir dispersion relation and have an average frequency which is approximately 11% higher than the average frequency of the localized field structures. The observations are consistent with the predictions of strong Langmuir turbulence theory. PMID- 11017343 TI - Measurements of energetic proton transport through magnetized plasma from intense laser interactions with solids AB - Protons with energies up to 18 MeV have been measured from high density laser plasma interactions at incident laser intensities of 5x10(19) W/cm(2). Up to 10(12) protons with energies greater than 2 MeV were observed to propagate through a 125 &mgr;m thick aluminum target and measurements of their angular deflection were made. It is likely that the protons originate from the front surface of the target and are bent by large magnetic fields which exist in the target interior. To agree with our measurements these fields would be in excess of 30 MG and would be generated by the beam of fast electrons which is also observed. PMID- 11017344 TI - Long-scale jet formation with specularly reflected light in ultraintense laser plasma interactions AB - Long-scale jetlike x-ray emission was observed in a 100-TW laser-plasma interaction. The jet was well collimated with a divergence of 30-40 mrad and continued from the target surface into underdense regions for a distance over 4 mm in the specular direction of the laser light. A two-dimensional particle-in cell simulation shows an electron acceleration with the specularly reflected laser light and collimation of the electron stream by a self-generated magnetic field, resulting in the electron jet to the direction of the specularly reflected light. PMID- 11017345 TI - Flow-induced beam steering in a single laser hot spot AB - The transmitted angular distribution of a 527 nm nearly diffraction-limited laser is measured after it propagates through a plasma with supersonic transverse flow. The laser beam is deflected by as much as 10 degrees and exhibits bowlike features in the flow direction, which is attributed to flow-induced beam steering. The finite interaction volume allows for direct comparison with a 3D hydrodynamic simulation, which is in good agreement with details of the experiment. PMID- 11017347 TI - Dynamic behavior of nickel atoms in graphitic networks AB - The dynamic behavior of nickel atoms in graphitic carbon onions, observed by in situ atomic-resolution electron microscopy, shows the formation of stable new C Ni phases. Nickel is observed to take substitutional in-plane positions in graphene layers, forming a planar graphitelike C-Ni lattice. Evidence is furthermore seen for the formation of a cubic C-Ni phase, suggesting a possible phase transformation in C-Ni from a graphitelike to a diamondlike structure. The stability of the planar phases is shown by first-principles calculations which also indicate that the C-Ni planes are metallic. PMID- 11017346 TI - Ab initio molecular dynamics with a classical pressure reservoir: simulation of pressure-induced amorphization in a Si35H36 cluster AB - We present a new constant-pressure ab initio molecular dynamics method suitable for studying, e.g., pressure-induced structural transformations in finite nonperiodic systems such as clusters. We immerse an ab initio treated cluster into a model classical liquid, described by a soft-sphere potential, which acts as a pressure reservoir. The pressure is varied by tuning the parameter of the liquid potential. We apply the method to a Si35H36 cluster, which undergoes a pressure-induced amorphization at approximately 35 GPa, and remains in a disordered state even upon pressure release. PMID- 11017348 TI - Hydrogen interaction with dislocations in Si AB - An H plasma has a remarkable effect on dislocation mobility in silicon, reducing its activation energy to 1.2 eV. Applying density functional theory to the interactions of H and H2 with the core of the 90 degrees partial dislocation in Si, we have identified a path for motion involving kink formation and migration at hydrogenated core bonds which conforms exactly to the experimentally measured activation energy. PMID- 11017349 TI - Density functional for hard sphere crystals: A fundamental measure approach AB - A new free energy density functional for hard spheres is presented, along the lines of the fundamental measure theory, which reproduces the Percus-Yevick equation of state and direct correlation function for the fluid, with a tensor weighted density. The functional, based on the zero-dimension limit, is exact for any one-dimensional density distribution of the spheres. The application to the hard sphere crystals gives excellent results, solving all of the qualitative problems of previous density functional approximations, including the unit cell anisotropy in the fcc lattice and the description of the metastable bcc lattice. PMID- 11017350 TI - Surface supercooling and stability of n-alkane films AB - The surface tension of n-octadecane was studied in the vicinity of the bulk melting point using both the maximum bubble pressure and Wilhelmy plate methods. The bubble surfaces were found to be supercooled below the surface freezing point. The onset of surface freezing is indicated by a sharp drop in surface tension at a constant temperature. This transition is accompanied by an increased film stability resulting in longer bubble lifetimes at the liquid surface. Variations in bubble lifetime reflect changes in the interfacial mechanical properties of the film from liquidlike to solidlike. PMID- 11017351 TI - Novel SiGe island coarsening kinetics: ostwald ripening and elastic interactions AB - Real-time light scattering measurements of coherent island coarsening during SiGe/Si heteroepitaxy reveal unusual kinetics. In particular, the mean island volume increases superlinearly with time, while the areal density of islands decreases at a faster-than-linear rate. Neither observation is consistent with standard considerations of Ostwald ripening. Modification of the standard theory to incorporate the effect of elastic interactions in the growing island array reproduces the observed behavior. PMID- 11017352 TI - Oxygen molecule dissociation on the Al(111) surface AB - The dissociative adsorption of the O2 on the Al(111) surface is studied using ab initio calculations based on density-functional theory. In the sticking probability experiments an activation barrier for the O2 dissociation exists, but our calculations predict a barrier only for one trajectory. Also our results do not support the model of charge transfer from the surface to the molecule as a bond breaking mechanism. Instead, the increasing hybridization between O2 orbitals and Al states, when the adsorbate approaches the slab, is significant for the dissociation. PMID- 11017353 TI - Calculation of polarization using a density functional method with localized charge AB - A density functional method, which represents the total charge density as a sum of self-consistently determined localized densities, is described. While this approach is generally less accurate than conventional band-structure methods, it offers a relatively simple interpretation of polarization and related properties. The method is illustrated with results for NaCl, MgO, and AlP. PMID- 11017354 TI - Anomalous magneto-oscillations in two-dimensional systems AB - The frequencies of Shubnikov-de Haas oscillations have long been used to measure the unequal population of spin-split two-dimensional subbands in inversion asymmetric systems. We report self-consistent numerical calculations and experimental results which indicate that these oscillations are not simply related to the zero-magnetic-field spin-subband densities. PMID- 11017355 TI - Electron spin density distribution in the polymer phase of CsC60: assignment of the NMR spectrum. AB - We present high resolution 133Cs-13C double resonance NMR data and 13C-13C NMR correlation spectra of 13C enriched samples of the polymeric phase of CsC60. These data lead to a partial assignment of the lines in the 13C NMR spectrum of CsC60 to the carbon positions on the C60 molecule. A plausible completion of the assignment can be made on the basis of an ab initio calculation. The data support the view that the conduction electron density is concentrated at the C60 "equator," away from the interfullerene bonds. PMID- 11017356 TI - Molecular geometry fluctuation model for the mobility of conjugated polymers. AB - We present a model to describe electrical transport in dense films of conjugated polymers. The essential physical features of the model are as follows: (i) thermal fluctuations in the molecular geometry of the polymer modify the energy levels of localized electronic states in the material, and (ii) the primary restoring force for these fluctuations is steric, which leads to spatial correlation in the energies of the localized electronic states. The model describes the electric field dependence of the mobility and explains the carrier density dependences of mobility observed in polymer diodes and field effect transistors. PMID- 11017357 TI - Canted antiferromagnetic phase in a double quantum well in a tilted quantizing magnetic field. AB - We investigate the double-layer electron system in a parabolic quantum well at filling factor nu=2 in a tilted magnetic field using capacitance spectroscopy. The competition between two ground states is found at the Zeeman splitting appreciably smaller than the symmetric-antisymmetric splitting. Although at the transition point the system breaks up into domains of the two competing states, the activation energy turns out to be finite, signaling the occurrence of a new insulator-insulator quantum phase transition. We interpret the obtained results in terms of a predicted canted antiferromagnetic phase. PMID- 11017358 TI - Landau-level spectroscopy of a two-dimensional electron system by tunneling through a quantum dot. AB - A single InAs self-assembled quantum dot is incorporated in the barrier of a tunnel diode and used as a spectroscopic probe of an adjacent two-dimensional electron system from the Fermi energy to the subband edge. We obtain quantitative information about the energy dependence of the quasiparticle lifetime. For magnetic field B, applied parallel to the current, we observe peaks in the current-voltage characteristics I(V) corresponding to the formation of Landau levels. Close to filling factor nu=1 we observe directly the exchange enhancement of the Lande g factor. PMID- 11017359 TI - Inverted electron-hole alignment in InAs-GaAs self-assembled quantum dots. AB - New information on the electron-hole wave functions in InAs-GaAs self-assembled quantum dots is deduced from Stark effect spectroscopy. Most unexpectedly it is shown that the hole is localized towards the top of the dot, above the electron, an alignment that is inverted relative to the predictions of all recent calculations. We are able to obtain new information on the structure and composition of buried quantum dots from modeling of the data. We also demonstrate that the excited state transitions arise from lateral quantization and that tuning through the inhomogeneous distribution of dot energies can be achieved by variation of electric field. PMID- 11017360 TI - Fluctuation conductivity in insulator-superconductor transitions with dissipation. AB - We analyze here the fluctuation conductivity in the vicinity of the critical point in a 2D Josephson junction array shunted by an Ohmic resistor. We find that, at the Gaussian level, the conductivity acquires a logarithmic dependence on T/m ( m is the inverse correlation length) when the dissipation is sufficiently small. In the renormalized classical regime, this logarithmic dependence gives rise to a leveling off of the resistivity at low to intermediate temperatures when fluctuations are included. We show, however, that this trend does not persist to T=0 at which point the resistivity vanishes. The possible relationship of the leveling of the resistivity to the low temperature transport in granular superconductors is discussed. PMID- 11017361 TI - Discrete breathers in nonlinear lattices: experimental detection in a josephson array. AB - We present the experimental detection of discrete breathers in an underdamped Josephson-junction array. Breathers exist under a range of dc current biases and temperatures, and are detected by measuring dc voltages. We find that the maximum allowable bias current for the breather is proportional to the array depinning current, while the minimum current seems to be related to a junction retrapping mechanism. We have observed that this latter instability leads to the formation of multisite breather states in the array. We have also studied the domain of existence of the breather at different values of the array parameters by varying the temperature. PMID- 11017362 TI - Observation of breathers in Josephson ladders. AB - We report on the observation of spatially localized excitations in a ladder of small Josephson junctions. The excitations are whirling states which persist under a spatially homogeneous force due to the bias current. These states of the ladder are visualized using a low temperature scanning laser microscopy. We also compute breather solutions with high accuracy in corresponding model equations. The stability analysis of these solutions is used to interpret the measured patterns in the I-V characteristics. PMID- 11017363 TI - Three-dimensional Josephson-junction arrays in the quantum regime. AB - We study the quantum phase transition properties of a three-dimensional periodic array of Josephson junctions with charging energy that includes both the self and mutual junction capacitances. We use the phase fluctuation algebra between number and phase operators, given by the Euclidean group E2, and we effectively map the problem onto a solvable quantum generalization of the spherical model. We obtain a phase diagram as a function of temperature, Josephson coupling, and charging energy. We also analyze the corresponding fluctuation conductivity and its universal scaling form in the vicinity of the zero-temperature quantum critical point. PMID- 11017364 TI - Order out of disorder in a gas of elastic quantum strings in 2+1 dimensions. AB - A limiting case of a dynamical stripe state which is of potential significance to cuprate superconductors is considered: a gas of elastic quantum strings in 2+1 dimensions, interacting merely via a hard-core condition. It is demonstrated that this gas always solidifies, by a mechanism which is the quantum analog of the entropic interactions known from soft condensed matter physics. PMID- 11017365 TI - Magnetotransport in manganites and the role of quantal phases: theory and experiment. AB - While low-temperature Hall resisitivity rhoxy of La2/3(Ca,Pb)1/3MnO3 single crystals can be separated into ordinary (OHE) and anomalous (AHE) contributions, no such decomposition is possible near the Curie temperature Tc. Rather, the rhoxy data collapse to a single function of the reduced magnetization m=M/Msat, with an extremum at approximately 0.4 m. A new mechanism for the AHE in the inelastic hopping regime is identified that reproduces the scaling curve. An extension of Holstein's model for the hopping OHE, the mechanism arises from the combined effects of the double-exchange-induced quantal phase in triads of Mn ions and spin-orbit interactions. PMID- 11017366 TI - 55Mn NMR investigation of electronic phase separation in La1-xCaxMnO3 for 0.2 < or = x < or = 0.5. AB - 55Mn NMR line shape measurements in La1-xCaxMnO3 for 0.20< or =x< or =0.50 provide experimental evidence about the existence of two distinct regions in the T-x magnetic phase diagram, where the homogeneous ferromagnetic (FM) metallic state is separated into FM metallic and FM insulating regions. These results are in agreement with recent theoretical predictions, which reveal a novel electronic phase separation in two FM states, providing orbital ordering and Jahn-Teller phonons are taken into consideration. PMID- 11017367 TI - Asymmetry in elementary events of magnetization reversal in a ferromagnetic/antiferromagnetic bilayer. AB - Real-time magneto-optical indicator film images reveal distinct asymmetry in the motion of a single domain wall in a wedged-NiFe/uniform-FeMn bilayer due to the nucleation and behavior of an exchange spring in the antiferromagnetic layer. Magnetization reversal from the ground state begins at the thick end of the wedge where the exchange anisotropy field (HE) is minimal and the magnetostatic field (HMS) is maximal, whereas reversal into the ground state begins from the thin end where HE is maximal and HMS is minimal. PMID- 11017368 TI - Photoinduced dehydrogenation of defects in undoped a-si:H using positron annihilation spectroscopy. AB - We report changes in variable-energy positron annihilation spectroscopy measurements on undoped hydrogenated amorphous silicon films after light soaking. The change, seen predominantly in the high momentum band of the annihilation radiation, is not reversed by thermal annealing. We suggest, following recent models of the Staebler-Wronski effect, that light exposure induces hydrogen trapped in vacancylike defects to become mobile in the Si network. The observations place constraints on models of hydrogen motion fitting macroscopic Staebler-Wronski effect kinetics and may help to achieve a definitive description of metastability in a-Si:H. PMID- 11017369 TI - Direct measurement of valence-charge asymmetry by x-ray standing waves. AB - By monitoring valence-photoelectron emission under condition of strong x-ray Bragg reflection, we have determined that a majority of GaAs valence charge resides on the anion sites of this heteropolar crystal, in quantitative agreement with the GaAs bond polarity as calculated from the Hartree-Fock term values. In contrast, the valence-charge distribution in Ge is found to be symmetric. In both cases, the valence emission is found to be closely coupled to the atomic cores. PMID- 11017370 TI - Effect of noble gases on sonoluminescence temperatures during multibubble cavitation. AB - Sonoluminescence spectra were collected from Cr(CO)6 solutions in octanol and dodecane saturated with various noble gases. The emission from excited-state metal atoms serves as an internal thermometer of cavitation. The intensity and temperature of sonoluminescence increases from He to Xe. The intensity of the underlying continuum, however, grows faster with increasing temperature than the line emission. Dissociation of solvent molecules within the bubble consumes a significant fraction of the energy generated by the collapsing bubble, which can limit the final temperature inside the bubble. PMID- 11017371 TI - Liquid-crystalline phase behavior of a colloidal rod-plate mixture. AB - The phase behavior of rod-plate mixtures was investigated using model systems containing unambiguously rod- and plate-shaped colloids. We find that the theoretically disputed biaxial nematic phase is unstable with respect to demixing into an isotropic and two uniaxial nematic phases. The phase behavior at very high densities is exceptionally rich and includes the coexistence of up to four different liquid crystalline phases, which stem from the coupling between the employed particle shapes and polydispersity. PMID- 11017372 TI - Absence of scaling for the intermediate scattering function of a hard-sphere suspension: static and dynamic x-ray scattering from concentrated polystyrene latex spheres. AB - X-ray photon correlation spectroscopy and small-angle scattering measurements are presented of the dynamics and structure of concentrated suspensions of charge stabilized polystyrene latex spheres dispersed in glycerol, for volume fractions from 3% to 52%. The static structures of the suspensions show essentially hard sphere behavior, and the short-time dynamics shows good agreement with predictions for the wave-vector-dependent collective diffusion coefficient. However, the intermediate scattering function is found to violate a scaling behavior found previously for a sterically stabilized hard-sphere suspension. PMID- 11017373 TI - Quantum and classical game strategies. PMID- 11017375 TI - Inequalities for dealing with detector inefficiencies in greenberger-horne zeilinger-type experiments AB - In this article we show that the three-particle Greenberger-Horne-Zeilinger theorem can be reformulated in terms of inequalities, allowing imperfect correlations due to detector inefficiencies. We show quantitatively that taking into account these inefficiencies, the published results of the Innsbruck experiment support the nonexistence of local hidden variables that explain the experimental results. PMID- 11017376 TI - Decoherence via the dynamical casimir effect AB - We derive a master equation for a mirror interacting with the vacuum field via radiation pressure. The dynamical Casimir effect leads to decoherence of a superposition state in a time scale that depends on the degree of "macroscopicity" of the state components, and which may be much shorter than the relaxation time scale. Coherent states are selected by the interaction as pointer states. PMID- 11017377 TI - Quantum time evolution in terms of nonredundant probabilities AB - Each scheme of state reconstruction comes down to parametrize the state of a quantum system by expectation values or probabilities directly measurable in an experiment. It is argued that the time evolution of these quantities provides an unambiguous description of the quantal dynamics. This is shown explicitly for a single spin s, using a quorum of expectation values which contains no redundant information. The quantum mechanical time evolution of the system is rephrased in terms of a closed set of linear first-order differential equation coupling (2s+1)(2) expectation values. This new representation of the dynamical law refers neither to the wave function of the system nor to its statistical operator. PMID- 11017378 TI - Vortex formation in a stirred bose-einstein condensate AB - Using a focused laser beam we stir a Bose-Einstein condensate of 87Rb confined in a magnetic trap and observe the formation of a vortex for a stirring frequency exceeding a critical value. At larger rotation frequencies we produce states of the condensate for which up to four vortices are simultaneously present. We have also measured the lifetime of the single vortex state after turning off the stirring laser beam. PMID- 11017379 TI - Surface excitations of a bose-einstein condensate AB - Surface modes in a Bose-Einstein condensate of sodium atoms have been studied. We observed excitations of standing and rotating quadrupolar and hexadecapolar modes. The modes were excited with high spatial and temporal resolution using the optical dipole force of a rapidly scanning laser beam. This novel technique is very flexible and should be useful for the study of rotating Bose-Einstein condensates and vortices. PMID- 11017380 TI - Yang-Lee theory for a nonequilibrium phase transition AB - To analyze phase transitions in a nonequilibrium system, we study its grand canonical partition function as a function of complex fugacity. Real and positive roots of the partition function mark phase transitions. This behavior, first found by Yang and Lee under general conditions for equilibrium systems, can also be applied to nonequilibrium phase transitions. We consider a one-dimensional diffusion model with periodic boundary conditions. Depending on the diffusion rates, we find real and positive roots and can distinguish two regions of analyticity, which can be identified with two different phases. In a region of the parameter space, both of these phases coexist. The condensation point can be computed with high accuracy. PMID- 11017381 TI - Noise induced stability in fluctuating, bistable potentials AB - The overdamped motion of a Brownian particle in an asymmetric, bistable, fluctuating potential shows noise induced stability: For intermediate fluctuation rates the mean occupancy of minima with an energy above the absolute minimum is enhanced. The model works as a detector for potential fluctuations being not too fast and not too slow. This effect occurs due to the different time scales in the problem. We present a detailed analysis of this effect using the exact solution of the Fokker-Planck equation for a simple model. Further we show that for not too fast fluctuations the system can be well described by effective rate equations. The results of the rate equations agree quantitatively with the exact results. PMID- 11017382 TI - Relativistic effects of light in moving media with extremely low group velocity AB - A moving dielectric medium acts as an effective gravitational field on light. One can use media with extremely low group velocities [Lene Vestergaard Hau et al., Nature (London) 397, 594 (1999)] to create dielectric analogs of astronomical effects on Earth. In particular, a vortex flow imprints a long-ranging topological effect on incident light and can behave like an optical black hole. PMID- 11017383 TI - Resonant production of topological defects AB - We describe a novel phenomenon in which vortices are produced due to resonant oscillations of a scalar field which is driven by a periodically varying temperature T, with T remaining much below the critical temperature T(c). Also, in a rapid heating of a localized region to a temperature below T(c), far separated vortex and antivortex can form. We compare our results with recent models of defect production during reheating after inflation. We also discuss possible experimental tests of our predictions of topological defect production without ever going through a phase transition. PMID- 11017384 TI - Observation of radiative leptonic decay of the tau lepton AB - Using 4.68 fb(-1) of e(+)e(-) annihilation data collected with the CLEO II detector at the Cornell Electron Storage Ring, we have studied tau radiative decays tau(-)-->nu(tau)&mgr;(-)nu;(&mgr;)gamma and tau(-)-->nu(tau)e( )nu;(e)gamma. For a 10 MeV minimum photon energy in the tau rest frame, the branching fraction for radiative tau decay to a muon or electron is measured to be (3.61+/-0.16+/-0. 35)x10(-3) or (1.75+/-0.06+/-0.17)x10(-2), respectively. The branching fractions are in agreement with standard model theoretical predictions. PMID- 11017385 TI - Search for a fourth-generation quark more massive than the Z0 boson in p&pmacr; collisions at radicals = 1.8 TeV AB - We present the results of a search for pair production of a fourth-generation charge -1 / 3 quark (b(')) in sqrt[s] = 1.8 TeV p&pmacr; collisions using 88 pb( 1) of data obtained with the Collider Detector at Fermilab. We assume that both quarks decay via the flavor-changing neutral current process b(')-->bZ(0) and that the b(') mass is greater than m(Z)+m(b). We studied the decay mode b(')b('); ->Z(0)Z(0)b&bmacr; where one Z0 decays into e(+)e(-) or &mgr;(+)&mgr;(-) and the other decays hadronically, giving a signature of two leptons plus jets. An upper limit on the sigma(p&pmacr;-->b(')b(');)x[B(b(')-->bZ(0))](2) is established as a function of the b(') mass. We exclude at 95% confidence level a b(') quark with mass between 100 and 199 GeV/c(2) for B(b(')-->bZ(0)) = 100%. PMID- 11017386 TI - Quark dispersion relation and dilepton production in the quark-gluon plasma AB - Under very general assumptions we show that the quark dispersion relation in the quark-gluon plasma is given by two collective branches, of which one has a minimum at a nonvanishing momentum. This general feature of the quark dispersion relation leads to structures (van Hove singularities, gaps) in the low mass dilepton production rate, which might provide a unique signature for the quark gluon plasma formation in relativistic heavy ion collisions. PMID- 11017387 TI - Transverse momentum and total cross section of e(+)e(-) pairs in the Z-boson region from p&pmacr; collisions at sqrt AB - The transverse momentum and total cross section of e(+)e(-) pairs in the Z-boson region of 6611.5 Gyr, the limit is 0.9 eV. PMID- 11017448 TI - Precision measurement of cosmic-Ray antiproton spectrum AB - The energy spectrum of cosmic-ray antiprotons ( &pmacr;'s) has been measured in the range 0.18-3.56 GeV, based on 458 &pmacr;'s collected by BESS in a recent solar-minimum period. We have detected for the first time a characteristic peak at 2 GeV of &pmacr;'s originating from cosmic-ray interactions with the interstellar gas. The peak spectrum is reproduced by theoretical calculations, implying that the propagation models are basically correct and that different cosmic-ray species undergo a universal propagation. Future BESS data with still higher statistics will allow us to study the solar modulation and the propagation in detail and to search for primary &pmacr; components. PMID- 11017450 TI - Shell structure of nuclei in strong magnetic fields in neutron star crusts AB - By employing Strutinsky's treatment we demonstrate that the magnetic field gives rise to a phase shift of the shell oscillations in nuclear masses. The proton orbital magnetism is shown to enhance the nuclear shell effect especially when the field influence is comparable to the spin-orbit coupling. The magnetic field of the strength scale B approximately 10(16)-10(17) G is found to shift significantly the nuclear magic numbers of the iron region towards smaller mass numbers. PMID- 11017451 TI - Nonlinear supergravity on a brane without compactification AB - We show that smooth domain wall spacetimes supported by a scalar field separating two anti-de Sitter-like regions admit a single graviton bound state. Our analysis yields a fully nonlinear supergravity treatment of the Randall-Sundrum model. Our solutions describe a pp-wave propagating in the domain wall background spacetime. If the latter is a Bogomol'nyi-Prasad-Sommerfeld state, our solutions retain some supersymmetry. Nevertheless, the Kaluza-Klein modes generate " pp curvature" singularities in the bulk located where the horizon of the anti-de Sitter region would ordinarily be. PMID- 11017452 TI - Tiling the plane without supersymmetry AB - We present a way of tiling the plane with a regular hexagonal network of defects. The network is stable and follows in consequence of the three-junctions that appear in a model of two real scalar fields that presents Z3 symmetry. The Z3 symmetry is effective in both the vacuum and defect sectors, and no supersymmetry is required to build the network. PMID- 11017453 TI - CPT and lorentz tests with muons AB - Precision experiments with muons are sensitive to Planck-scale CPT and Lorentz violation that is undetectable in other tests. Existing data on the muonium ground-state hyperfine structure and on the muon anomalous magnetic moment could be analyzed to provide dimensionless figures of merit for CPT and Lorentz violation at the levels of 4x10(-21) and 10(-23). PMID- 11017454 TI - Meson structure in a relativistic many-body approach AB - Results from an extensive relativistic many-body analysis utilizing a realistic effective QCD Hamiltonian are presented for the meson spectrum. A comparative numerical study of the BCS, Tamm-Dancoff (TDA), and RPA treatments provides new, significant insight into the condensate structure of the vacuum, the chiral symmetry governance of the pion, and the meson spin, orbital, and flavor mass splitting contributions. In contrast to a previous glueball application, substantial quantitative differences are computed between TDA and RPA for the light quark sector with the pion emerging as a Goldstone boson only in the RPA. PMID- 11017455 TI - Parity violation in elastic electron-proton scattering and the Proton's strange magnetic form factor AB - We report a new measurement of the parity-violating asymmetry in elastic electron scattering from the proton at backward scattering angles. This asymmetry is sensitive to the strange magnetic form factor of the proton as well as electroweak axial radiative corrections. The new measurement of A = -4.92+/ 0.61+/-0.73 ppm provides a significant constraint on these quantities. The implications for the strange magnetic form factor are discussed in the context of theoretical estimates for the axial corrections. PMID- 11017456 TI - Search for color singlet technicolor particles in p&pmacr; collisions at radicals = 1.8 TeV AB - We search for color singlet technirho and technipion production in p&pmacr; collisions at sqrt[s] = 1.8 TeV recorded with the Collider Detector at Fermilab. These exotic technimesons are present in a model of walking technicolor. The signatures studied are lepton plus two jets plus E(T) and multijet final states. No excess of events is seen in either final state. We set an upper limit on the technirho production cross section and exclude a region in the technipion mass versus technirho mass plane. PMID- 11017457 TI - Discovery of doubly magic 48Ni AB - In an experiment at the SISSI/LISE3 facility of GANIL, we used the projectile fragmentation of a primary 58Ni26+ beam at 74.5 MeV/nucleon with an average current of 3 &mgr;A on a natural nickel target to produce very neutron-deficient isotopes. In a 10-day experiment, 287 42Cr isotopes, 53 45Fe isotopes, 106 49Ni isotopes, and 4 48Ni isotopes were unambiguously identified. The doubly magic nucleus 48Ni, observed for the first time, is the most proton-rich isotope ever identified with an isospin projection T(z) = -4. It is probably the last doubly magic nucleus with "classical" shell closures accessible for present-day facilities. Its observation allows us to deduce a lower limit for the half-life of 48Ni of 0.5 &mgr;s. PMID- 11017458 TI - Isospin tracing: A probe of nonequilibrium in central heavy-Ion collisions AB - Four different combinations of 9644Ru and 9640Zr nuclei, both as projectile and target, were investigated at the same bombarding energy of 400A MeV using a 4pi detector. The degree of isospin mixing between projectile and target nucleons is mapped across a large portion of the phase space using two different isospin tracer observables, the number of measured protons and the tritium to 3He yield ratio. The experimental results show that the global equilibrium is not reached even in the most central collisions. Quantitative measures of stopping and mixing are extracted from the data. They are found to exhibit a quite strong sensitivity to the in-medium ( n,n) cross section used in microscopic transport calculations. PMID- 11017459 TI - Guiding neutral atoms on a chip AB - We demonstrate the guiding of neutral atoms by the magnetic fields due to microfabricated current-carrying wires on a chip. Atoms are guided along a magnetic field minimum parallel to and above the current-carrying wires. Two guide configurations are demonstrated: one using two wires with an external magnetic field, and a second using four wires without an external field. These guide geometries can be extended to integrated atom optics circuits, including beam splitters. PMID- 11017460 TI - Stability of the ground state vinylidene anion H2CC- AB - A technique to measure very low collisionless decay rates of ionic species has been applied at a new electrostatic storage ring to obtain the rate for spontaneous rearrangement of the vinylidene anion to neutral acetylene. The measurement yields a rate of k(0) = (0.009+/-0.006) s(-1), corresponding to a natural lifetime of the vinylidene anion of tau(0) approximately 110 s. PMID- 11017461 TI - Ultrafast hot-electron dynamics observed in Pt( -)(3) using time-resolved photoelectron spectroscopy AB - Time-resolved two-photon photoelectron spectra have been measured for free Pt( )(3) using femtosecond pulses of 1.5 eV photon energy in a pump-probe configuration. The time-dependent photoelectron distribution reveals a lifetime of optically excited states of less than 70 fs. Such an unexpected fast electron relaxation in Pt( -)(3) suggests the existence of inelastic electron-electron scattering processes in a triatomic cluster which result in a lifetime similar to those of bulk metals. PMID- 11017462 TI - Measurement of the 1s-2s energy interval in muonium AB - The 1s-2s interval has been measured in the muonium (&mgr;(+)e(-)) atom by Doppler-free two-photon pulsed laser spectroscopy. The frequency separation of the states was determined to be 2 455 528 941.0(9.8) MHz, in good agreement with quantum electrodynamics. The result may be interpreted as a measurement of the muon-electron charge ratio as -1-1.1(2.1)x10(-9). We expect significantly higher accuracy at future high flux muon sources and from cw laser technology. PMID- 11017463 TI - Double tunneling: An overlooked quantum effect in anionic molecular clusters AB - Simultaneous tunneling of an electron and nuclei in hydrogen-bonded molecular clusters which support a dipole bound electron is reported. A whole class of systems including hydrogen halide and water dimer anions is predicted to exhibit this effect. A measurable signature of double tunneling is a strong reduction of the tunneling splitting compared to the neutral cluster. Quantum mechanical calculations give for the hydrogen fluoride dimer anion a ground-state tunneling splitting of 114 MHz, while for the first excited vibrational state the tunneling splitting reaches 4.20 GHz. PMID- 11017464 TI - Variational study of 3He droplets AB - We report variational calculations of energies of (3)He(N) droplets ( 20, would vanish unless i = i('), as in the case of a homogeneous ensemble in which the i's represent wave vectors. We analytically treat an extension to the case in which one has a larger number of coefficients than realizations, introducing intrinsic linear dependences that vitiate the stated condition. PMID- 11017473 TI - The D'yakov-kontorovich instability of shock waves in real gases AB - In the 1950s, D'yakov and Kontorovich predicted that under certain conditions perturbed shock waves in nonideal gases can become unstable by emitting undamped sound and entropy-vortex waves. For the last 45 years, though, little progress has been made in the identification and numerical modeling of physical conditions for which this phenomenon might occur. Using a van der Waals equation of state, we present for the first time a dynamical simulation of a D'yakov-Kontorovich instability. The two-dimensional emission pattern of acoustic waves appearing in the simulation agrees with the prediction of a linearized theory. PMID- 11017474 TI - Observation of a hybrid spin resonance AB - A new type of spin depolarization resonance has been observed at the Brookhaven Alternating Gradient Synchrotron (AGS). This spin resonance is identified as a strong closed-orbit sideband around the dominant intrinsic spin resonance. The strength of the resonance was proportional to the 9th harmonic component of the horizontal closed orbit and proportional to the vertical betatron oscillation amplitude. This "hybrid" spin resonance cannot be overcome by the partial snake at the AGS, but it can be corrected by the harmonic orbit correctors. PMID- 11017476 TI - Observation of avalanchelike phenomena in a magnetically confined plasma AB - Electron temperature fluctuations seen in a magnetically confined tokamak plasma have some of the characteristics of the avalanchelike events sometimes associated with self-organized criticality, including intermittency, large space and time scales, " 1/f" spectra, large tails in the autocorrelation function, and clear evidence of radial propagation. PMID- 11017475 TI - Free-electron laser as a driver for a resonant cavity at 35 GHz AB - An intense beam of relativistic electrons (800 A, 6.7 MeV) has been bunched at 35 GHz by a free-electron laser, in which output power levels exceeding 100 MW were obtained. The beam was then extracted and transported through a resonant cavity, which was excited by its passage. Microwave power levels of 10 MW were extracted from the cavity, in reasonable agreement with the simple formula which relates power to known properties of both the beam and the cavity. PMID- 11017477 TI - Helicity redistribution during relaxation of astrophysical plasmas AB - We present the first 3D numerical MHD simulations that show that Taylor's relaxation conjecture is not satisfied in some MHD evolution of magnetic configurations encountered in solar physics. We show that magnetic helicity can be slowly injected through the boundary into a magnetic configuration which then evolves into a MHD disruption, with the formation in finite time of a current sheet through which reconnection occurs, leading to a release of magnetic energy. While helicity is well conserved during the process, it is shown that the relaxed state is far from the constant- alpha linear force-free field that would be predicted by Taylor's conjecture. PMID- 11017478 TI - Ion larmour radius effect on rf ponderomotive forces and induced poloidal flow in tokamak plasmas AB - Analytical approximations are used to clarify the effect of Larmour radius on rf ponderomotive forces and on poloidal flows induced by them in tokamak plasmas. The electromagnetic force is expressed as a sum of a gradient part and of a wave momentum transfer force, which is proportional to wave dissipation. The first part, called the gradient electromagnetic stress force, is combined with fluid dynamic (Reynolds) stress force, and gyroviscosity is included into viscosity force to model finite ion Larmour radius effects in the momentum response to the rf fields in plasmas. The expressions for the relative magnitude of different forces for kinetic Alfven waves and fast waves are derived. PMID- 11017479 TI - Sawtooth evolution during JET ion-cyclotron-resonance-heated pulses AB - The recent Joint European Torus deuterium-tritium campaign has yielded ion cyclotron-resonance-heated (ICRH) pulses during which both the sawtooth characteristics and the ICRH minority ion population both evolve substantially. At multiple times during each pulse, the evolution of the kinetic-fluid MHD energy is calculated from measurement of the energetic ions and compared with the evolving sawtooth duration. There is strong correlation between sawtooth duration and minority ion stabilization of the ideal internal kink. PMID- 11017480 TI - Detuned raman amplification of short laser pulses in plasma AB - The recently proposed scheme of so-called "fast compression" of laser pulses in plasma can increase peak laser intensities by 10(5) [Phys. Rev. Lett. 82, 4448 (1999)]. The compression mechanism is the transient stimulated Raman backscattering, which outruns the fastest filamentation instabilities of the pumped pulse even at highly overcritical powers. This Letter proposes a novel nonlinear filtering effect that suppresses premature backscattering of the pump in a noisy plasma layer, while the desired amplification of a sufficiently intense seed persists with a high efficiency. The effect is of basic interest and also makes it robust to noise the simplest technologically fast compression scheme. PMID- 11017481 TI - Nature of monster sawteeth and their relationship to Alfven instabilities in tokamaks AB - A correlation is explored between the presence of energetic particle modes (EPM) and long-period sawtooth oscillations in tokamak plasmas heated by rf waves. The eventual crash of these sawteeth is explained in terms of the loss of the stabilizing fast particles due to the EPM. The absence of long-period sawteeth in high q(a) discharges is explained in terms of ion loss due to toroidal Alfven eigenmodes. PMID- 11017482 TI - Energy confinement time and heat transport in initial neutral beam heated plasmas on the large helical device AB - The confinement characteristics of large net-current-free plasmas heated by neutral-beam injection have been investigated in the Large Helical Device (LHD). A systematic enhancement in energy-confinement times from the scaling derived from the medium-sized heliotron/torsatron experiments have been observed, which is attributed to the edge pedestal. The core confinement is scaled with the Bohm term divided by the square root of the gyro radii. The comparative analysis using a dimensionally similar discharge in the Compact Helical System indicates gyro Bohm dependence in the core and transport improvement in the edge region of LHD plasmas. PMID- 11017483 TI - Density-functional theory of inhomogeneous fluids in the canonical ensemble AB - We present a density-functional approach for dealing with inhomogeneous fluids in the canonical ensemble. A general relation is proposed between the free-energy functionals in the canonical and the grand canonical ensembles. The minimization of the canonical-ensemble free-energy functional gives rise to Euler-Lagrange equations which involve averaged Ornstein-Zernike equations of second and third order. The theory is especially appropriate for systems with a small, fixed number of particles. As an example of application we obtain accurate results for the density profile of a hard-sphere fluid in a closed spherical cavity that contains only a few particles. PMID- 11017484 TI - Librons observed in liquid acetonitrile by hyper-rayleigh scattering AB - Hyper-Rayleigh scattering intensity ratios I(VV)/I(VH) in the range 9-26 are observed for liquid acetonitrile (CH 3CN), inconsistent with localized modes of motion in an isotropic fluid. The observations are consistent with propagating orientational modes (librons) with frequency nu = 2 cm (-1), which scatter 71% of the light. PMID- 11017485 TI - Pseudo-casimir structural force drives spinodal dewetting in nematic liquid crystals AB - We analyze theoretically the fluctuation-induced force in thin nematic films subject to competing surface interactions, and we find that the force is attractive at small distances and repulsive otherwise. The results provide a consistent interpretation of a recent study of spinodal dewetting of 5CB on a silicon wafer [F. Vandenbrouck et al., Phys. Rev. Lett. 82, 2693 (1999)], implying that this experiment can be regarded as the first observation of the pseudo-Casimir effect in liquid crystals. PMID- 11017486 TI - Microscopic origin of the phenomenological equilibrium "Doping limit Rule" in n type III-V semiconductors AB - The highest equilibrium free-carrier doping concentration possible in a given material is limited by the "pinning energy" which shows a remarkable universal alignment in each class of semiconductors. Our first-principles total energy calculations reveal that equilibrium n-type doping is ultimately limited by the spontaneous formation of close-shell acceptor defects: the (3-)-charged cation vacancy in AlN, GaN, InP, and GaAs and the (1-)-charged DX center in AlAs, AlP, and GaP. This explains the alignment of the pinning energies and predicts the maximum equilibrium doping levels in different materials. PMID- 11017487 TI - Dynamics of Si-H vibrations in an amorphous environment AB - We present results of the first vibrational photon-echo, transient-grating, and temperature dependent transient-bleaching experiments on a-Si:H. Using these techniques, and the infrared light of a free electron laser, the vibrational population decay and phase relaxation of the Si-H stretching mode were investigated. Careful analysis of the data indicates that the vibrational energy relaxes directly into Si-H bending modes and Si phonons, with a distribution of rates determined by the amorphous host. Conversely, the pure dephasing appears to be single exponential, and can be modeled by dephasing via two-phonon interactions. PMID- 11017488 TI - Freezing by heating in a driven mesoscopic system AB - We investigate a simple model corresponding to particles driven in opposite directions and interacting via a repulsive potential. The particles move off lattice on a periodic strip and are subject to random forces as well. We show that this model-which can be considered as a continuum version of some driven diffusive systems-exhibits a paradoxical, new kind of transition called here "freezing by heating." One interesting feature of this transition is that a crystallized state with a higher total energy is obtained from a fluid state by increasing the amount of fluctuations. PMID- 11017489 TI - Topology, phase instabilities, and wetting of microemulsion networks AB - We predict theoretically the gradual formation of fluctuating, connected microemulsion networks from disconnected globules as the spontaneous curvature is varied, in agreement with recent direct measurements of these topological transitions. The connectivity induced instability together with emulsification failure of the network relate the ultralow tensions and wetting transition to the changing microstructure. PMID- 11017490 TI - Slow 4He quenches produce fuzzy, transient vortices AB - We examine the Zurek scenario for the production of vortices in quenches of liquid 4He in the light of recent experiments. Extending our previous results to later times, we argue that short wavelength thermal fluctuations make vortices poorly defined until after the transition has occurred. Further, if and when vortices appear, it is plausible that they will decay faster than anticipated from turbulence experiments, irrespective of quench rates. PMID- 11017491 TI - Repetitive single vortex-loop creation by a vibrating wire in superfluid 3He-B AB - Spectacular features are observed on the velocity-force characteristics of a vibrating wire resonator in superfluid 3He-B at ultralow temperatures. Both plateaus and discontinuities are seen in the characteristics. The plateaus seem to have two separate critical velocities where first some "event" occurs, which causes the wire to lose energy and slow down, followed by a second lower critical velocity where the event decouples. It is suggested that these events are due to vortex-loop creation at protuberances on the vibrating wire. This opens up the possibility of controlling the creation of vorticity through specially prepared protuberances. PMID- 11017492 TI - Ab initio modeling of metal adhesion on oxide surfaces with defects AB - Our ab initio studies show that surface defects cause redistribution of the electron density which can increase substantially the binding energy of metal atoms to oxide surfaces. The results for electron (F(0)(s)) and hole (V(0)(s)) centers in the adhesion of Ag atoms (at 1:4 and 1:1 coverages) to a MgO(100) surface, combined with previous studies for charged defects, support earlier ideas of the mechanism of radiation-enhanced adhesion of nonreactive metals on oxide substrates. The results suggest that some optical control of adhesion energies is possible through charge transfer. PMID- 11017493 TI - Multiscale simulation of loading and electrical resistance in silicon nanoindentation AB - Nanoindentation experiments are an excellent probe of micromechanical properties, but their interpretation is complicated by their multiscale nature. We report simulations of silicon nanoindentation, based on an extended version of the local quasicontinuum model, capable of handling complex Bravais crystals. Our simulations reproduce the experimental load vs displacement curves and provide microscopic information such as the distribution of transformed metallic phases of silicon underneath the indenter. This information is linked to the macroscopic electrical resistance, giving a satisfactory explanation of experimental results. PMID- 11017494 TI - Ferromagnetism in doped excitonic insulators AB - The theory of doped excitonic insulators is reinvestigated in light of recent experiments on hexaborides. For the appropriate valley-degenerate X3,X'3 band structure, "intravalley" condensation is energetically favored. Ferromagnetism occurs upon doping due to the quenching of kinetic energy at the otherwise direct first-order excitonic insulator-metal transition. The phase diagram includes states of spatially inhomogeneous density and magnetization at low temperatures. PMID- 11017495 TI - Virial coefficients of non-abelian anyons AB - We study a system of non-Abelian anyons in the lowest Landau level of a strong magnetic field. Using diagrammatic techniques, we prove that the virial coefficients do not depend on the statistics parameter. This is true for all representations of all non-Abelian groups for the statistics of the particles and relies solely on the fact that the effective statistical interaction is a traceless operator. PMID- 11017496 TI - Dynamical properties of the one-dimensional band insulator (NbSe4)3I AB - Optical and photoemission experiments reveal unexpected spectral signatures of one-dimensional band insulators. In the model compound (NbSe (4))3I the optical conductivity decays as a power law sigma(1)(omega) approximately omega(-4.25) above a sharp gap edge. Photoemission observes both the valence and a shadow band, produced by a commensurate superstructure. We identify an optical and photoemission band gap consistent with other measurements but much smaller than the energy scale defined by the dispersion of the band peak in the photoemission spectra. PMID- 11017497 TI - Metal-insulator transitions: influence of lattice structure, jahn-teller effect, and Hund's rule coupling AB - We study the influence of the lattice structure, the Jahn-Teller effect, and the Hund's rule coupling on a metal-insulator transition in A(n)C60 (A = K,Rb). The difference in the lattice structure favors A3C60 (fcc) being a metal and A4C60 (bct) being an insulator, and the coupling to H(g) Jahn-Teller phonons favors A4C60 being nonmagnetic. The coupling to H(g) ( A(g)) phonons decreases (increases) the value U(c) of the Coulomb integral at which the metal-insulator transition occurs. There is an important partial cancellation between the Jahn Teller effect and the Hund's rule coupling. PMID- 11017498 TI - Semiclassical theory of conductance and noise in open chaotic cavities AB - Conductance and shot noise of an open cavity with diffusive boundary scattering are calculated within the Boltzmann-Langevin approach. In particular, conductance contains a nonuniversal geometric contribution, originating from the presence of open contacts. Subsequently, universal expressions for multiterminal conductance and noise, valid for all chaotic cavities, are obtained classically, based on the fact that the distribution function in the cavity depends only on energy, and using the principle of minimal correlations. PMID- 11017499 TI - Bistability in scanning tunneling spectroscopy of Ga-terminated Si(111) AB - Bistable electron transport, a phenomenon usually associated with double-barrier structures, has been observed with a conventional STM junction formed between a metal tip and a Ga-terminated Si(111) surface at 77 K. Large hysteresis loops appear in the current-voltage characteristics when electrons are injected from the tip to the surface. The turn-on bias varies from -3.1 to -4.0 V and shows an inverse dependence on the tip-sample distance, indicating a strong field effect. The turn-off bias, however, is essentially pinned at a conductance threshold of 2.7 V. PMID- 11017500 TI - Finite-temperature density instability at high landau level occupancy AB - We study here the onset of charge density wave instabilities in quantum Hall systems at finite temperature for Landau level filling nu>4. Specific emphasis is placed on the role of disorder as well as on an in-plane magnetic field. Beyond some critical value, disorder is observed to suppress the charge density wave melting temperature to zero. In addition, we find that a transition from perpendicular to parallel stripes (relative to the in-plane magnetic field) exists when the electron gas thickness exceeds approximately 60 A. The perpendicular alignment of the stripes is in agreement with the experimental finding that the easy conduction direction is perpendicular to the in-plane field. PMID- 11017501 TI - Relevance of disorder for dirac fermions with imaginary vector potentials AB - We consider the effects of disorder in a Dirac-like Hamiltonian. In order to use conformal field theory techniques, we argue that one should consider disorder in an imaginary vector potential. This affects significantly the signs of the lowest order beta functions. We present evidence for the existence of two distinct universality classes, depending on the relative strengths of the gauge field verses impurity disorder strengths. In one class all disorder is made irrelevant by the gauge field disorder. PMID- 11017502 TI - Interlayer quasiparticle transport in the vortex state of josephson coupled superconductors AB - We calculate the dependence of the interlayer quasiparticle conductivity, sigma(q), in a Josephson coupled d-wave superconductor on the magnetic field B parallelc and the temperature T. We consider a clean superconductor with resonant impurity scattering and a dominant coherent interlayer tunneling. When pancake vortices in adjacent layers are weakly correlated, at low T the conductivity increases sharply with B over a field scale determined by the impurity scattering, before reaching an extended region of slow linear growth. At high T the conductivity initially decreases and then reaches the same linear regime. For correlated pancakes, sigma(q) increases much more strongly with the applied field. PMID- 11017503 TI - Observation of cascaded two-photon-induced transitions between fluxoid states of a SQUID AB - We present evidence for transitions between fluxoid wells of a SQUID due to cascaded, two-photon processes. Such transitions are evidenced by an anomalous dependence on the transition rate from the one-photon resonant level within the initial well, which cannot be explained by previously observed macroscopic resonant tunneling. These two-photon processes may be a significant source of decoherence in SQUID qubits subject to microwave radiation. PMID- 11017504 TI - Vortex entanglement in disordered superconductors AB - Vortex entanglement and pinning in multiply connected disordered superconductors are studied. It is shown that the winding of vortices around repulsive obstacles is greatly enhanced by quenched columnar disorder and suppressed by point disorder, compared to the clean case. This leads to an additional contribution to the effective pinning force acting on vortices, which, unlike the conventional mechanisms of pinning, grows with temperature. PMID- 11017505 TI - Exact dynamical structure factor of the degenerate haldane-shastry model AB - The dynamical structure factor S(q,omega) of the K-component ( K = 2, 3, 4) spin chain with a 1/r(2) interaction is derived exactly at zero temperature for the arbitrary size of the system. The result is interpreted in terms of a free quasiparticle picture which is a generalization of the spinon picture in the SU(2) case. The excited states consist of K quasiparticles each of which is characterized by a set of K-1 quantum numbers. Divergent singularities of S(q,omega) at the spectral edges are derived analytically. The analytic result is checked numerically for finite systems. PMID- 11017506 TI - Step-edge induced anisotropic domain-wall propagation AB - We report the observation of anisotropic domain-wall propagation in ultrathin magnetic films with perpendicular anisotropy. A controlled density of step edges was introduced which allowed us to quantify its influence on the domain-wall pinning. For a sawtooth arrangement of the step edges the corresponding wall movement resulted in triangular shaped domains. All aspects of this anisotropic domain-wall evolution could be reproduced by a simulation based on a modified Ginzburg-Landau-type soft-spin model. PMID- 11017507 TI - Dynamics of a domain wall in soft-magnetic materials: barkhausen effect and relation with sandpile models AB - The CZDE model [P. Cizeau, S Zapperi, G. Durin, and H. E. Stanley, Phys. Rev. Lett. 79, 4669 (1997)] for the dynamics of a domain wall in soft-magnetic materials is investigated. The equation of motion for the domain wall is reduced to a dimensionless form where the control parameters are clearly identified. In this way we show that in soft-magnetic materials with low anisotropies the noise can be approximated by a columnar disorder, and perturbation theory gives a good estimate of the avalanche exponents. Moreover, the resulting exponents are found to be identical to those obtained for directed Abelian sandpile models. The analogies and differences with these models and the question of self-organized criticality in the Barkhausen effect are discussed. PMID- 11017508 TI - Nuclear spin relaxation rates in two-Leg spin ladders AB - Using the transfer-matrix density-matrix renormalization group method, we study the nuclear spin relaxation rate 1/T(1) in the two-leg s = 1 / 2 ladder as a function of the interchain (J( perpendicular)) and intrachain (J( parallel)) couplings. In particular, we separate the q(y) = 0 and pi contributions and show that the latter contribute significantly to the copper relaxation rate (63)(1/T(1)) in the experimentally relevant coupling and temperature range. We compare our results to both theoretical predictions and experimental measures on ladder materials. PMID- 11017509 TI - Periodic resonance excitation and intertube interaction from quasicontinuous distributed helicities in single-wall carbon nanotubes AB - Photoselective resonance Raman scattering from laser ablation grown single-wall carbon nanotubes is demonstrated to be consistent with a response from tubes with all geometrically allowed helicities. This information is drawn from an analysis of the resonance scattering by combining ab initio calculations for the mode frequencies with evaluations of the resonance cross sections for isolated tubes. The resonance excitation was found to exhibit an oscillatory behavior. To match the experiments and the calculations, the frequencies obtained from the latter must be up-shifted by 8.5% on the average. This stiffening is ascribed to the tube-tube interaction in the carbon nanotube bundles. PMID- 11017510 TI - Microimplosions: cavitation collapse and shock wave emission on a nanosecond time scale AB - A streak camera with high spatial and temporal resolution was used for imaging the dynamics of the violent collapse in single-bubble sonoluminescence. The high pressure in the last phase of the bubble collapse leads to the emission of a shock wave, which is launched with a shock velocity of almost 4000 m/s. The shock amplitude decays much faster than approximately 1/r. From the strongly nonlinear propagation the pressure in the vicinity of the bubble can be calculated to be in the range of 40-60 kbar. PMID- 11017511 TI - Atom abstraction in the scattering of state-selected NO+(X1Sigma(+)) on O/Al(111) AB - The reactive scattering of state-selected NO+(X1Sigma(+)) on oxygen-covered Al(111) is explored at hyperthermal collision energies. Relative ion yields and mean translational energies of scattered NO-2 are presented as a function of oxygen exposure and NO+ collision energy. Above the 9+/-1 eV threshold for reaction, NO-2 products emerge with an average kinetic energy that depends linearly on incident NO+ energy. The formation of NO-2 is assigned to the direct, Eley-Rideal abstraction of an adsorbed O atom by an incident NO molecule. PMID- 11017512 TI - Coupling between flow and structure for a lamellar surfactant phase AB - The flow-structure relation of lamellar phases is studied using rheometry and cross-polarized microscopy under flow. The equilibrium phases show different defects. Low salinities lead to very viscous, "onion" phases, whereas at high salinity, a low viscosity plane lamellar phase is found. Under shear, the latter shows a sudden transition to a viscoelastic gel, with a texture and viscosity very similar to that of the onions. Gelation occurs after a certain delay time, increasing rapidly with salinity, by the nucleation of onions. This allows one to relate the delay time to the defect energy. PMID- 11017514 TI - A space-time adaptive method for simulating complex cardiac dynamics. AB - For plane-wave and many-spiral states of the experimentally based Luo-Rudy 1 model of heart tissue in large (8 cm square) domains, we show that a space-time adaptive time-integration algorithm can achieve a factor of 5 reduction in computational effort and memory-but without a reduction in accuracy-when compared to an algorithm using a uniform space-time mesh at the finest resolution. Our results indicate that such an algorithm can be extended straightforwardly to simulate quantitatively three-dimensional electrical dynamics over the whole human heart. PMID- 11017513 TI - Evidence for non-DLVO hydration interactions in solutions of the protein apoferritin. AB - We have studied molecular interactions in solutions of the protein apoferritin by static and dynamic light scattering. When plotted against the electrolyte concentration, the second osmotic virial coefficient exhibits a minimum. The ascending branch of this dependence is a manifestation of a surprisingly strong repulsion between the molecules at electrolyte concentrations about and above 0.2M, where electrostatic interactions are suppressed. We argue that the repulsion is due to the water structuring, enhanced by the accumulation of hydrophilic counterions around the apoferritin molecules, giving rise to so called hydration forces. PMID- 11017515 TI - Random costs in combinatorial optimization AB - The random cost problem is the problem of indentifying the minimum in a list of random numbers. By definition, this problem cannot be solved faster than by exhaustive search. It is shown that a classical NP-hard optimization problem, number partitioning, is essentially equivalent to the random cost problem. On the one hand this explains the bad performance of heuristic approaches to the number partitioning problem, but on the other hand it allows one to calculate the probability distributions of the optimum and suboptimum costs. PMID- 11017516 TI - From massively parallel algorithms and fluctuating time horizons to nonequilibrium surface growth AB - We study the asymptotic scaling properties of a massively parallel algorithm for discrete-event simulations where the discrete events are Poisson arrivals. The evolution of the simulated time horizon is analogous to a nonequilibrium surface. Monte Carlo simulations and a coarse-grained approximation indicate that the macroscopic landscape in the steady state is governed by the Edwards-Wilkinson Hamiltonian. Since the efficiency of the algorithm corresponds to the density of local minima in the associated surface, our results imply that the algorithm is asymptotically scalable. PMID- 11017517 TI - Typical performance of gallager-type error-correcting codes AB - The performance of Gallager's error-correcting code is investigated via methods of statistical physics. In this approach, the transmitted codeword comprises products of the original message bits selected by two randomly constructed sparse matrices; the number of nonzero row/column elements in these matrices constitutes a family of codes. We show that Shannon's channel capacity is saturated for many of the codes while slightly lower performance is obtained for others which may be of higher practical relevance. Decoding aspects are considered by employing the Thouless-Anderson-Palmer approach which is identical to the commonly used belief propagation-based decoding. PMID- 11017518 TI - Conformally invariant fractals and potential theory AB - The multifractal (MF) distribution of the electrostatic potential near any conformally invariant fractal boundary, like a critical O(N) loop or a Q-state Potts cluster, is solved in two dimensions. The dimension &fcirc;(straight theta) of the boundary set with local wedge angle straight theta is &fcirc;(straight theta) = pi / straight theta-25-c / 12 (pi-straight theta)(2) / straight theta(2pi-straight theta), with c the central charge of the model. As a corollary, the dimensions D(EP) of the external perimeter and D(H) of the hull of a Potts cluster obey the duality equation (D(EP)-1) (D(H)-1) = 1 / 4. A related covariant MF spectrum is obtained for self-avoiding walks anchored at cluster boundaries. PMID- 11017519 TI - "Weighing" a closed system and the time-energy uncertainty principle AB - A gedanken experiment is proposed for "weighing" the total mass of a closed system from within the system. We prove that for an internal observer the time tau, required to measure the total energy with accuracy DeltaE, is bounded according to tauDeltaE>Planck's over 2pi. This time-energy uncertainty principle for a closed system follows from the measurement backreaction on the system. We generally examine what other conserved observables are in principle measurable within a closed system and what are the corresponding uncertainty relations. PMID- 11017520 TI - Propagation of cold atoms along a miniature magnetic guide AB - A cloud of laser-cooled 85Rb atoms is coupled through a magnetic funnel into a miniature waveguide formed by four current-carrying wires embedded in a silica fiber. The atom cloud has a approximately 100 &mgr;m radius within the fiber and propagates over cm distances. We study the coupling, propagation, and transverse distribution of atoms in the fiber, and find good agreement with theory. This prototype demonstrates the feasibility of miniature guides as a tool in the new field of integrated atom optics, leading to single-mode propagation of de Broglie waves and the possible preparation of 1D atom clouds. PMID- 11017521 TI - Completely positive quantum dissipation AB - A completely positive master equation describing quantum dissipation for a Brownian particle is derived starting from microphysical collisions, exploiting a recently introduced approach to subdynamics of a macrosystem. The obtained equation can be cast into Lindblad form with a single generator for each Cartesian direction. Temperature dependent friction and diffusion coefficients for both position and momentum are expressed in terms of the collision cross section. PMID- 11017522 TI - Unstable superheavy relic particles as a source of neutrinos responsible for ultrahigh-energy cosmic rays AB - Decays of superheavy relic particles may produce extremely energetic neutrinos. Their annihilations on the relic neutrinos can be the origin of the cosmic rays with energies beyond the Greisen-Zatsepin-Kuzmin cutoff. The redshift acts as a cosmological filter selecting the sources at some particular value z(e)+/-deltaz, for which the present neutrino energy is close to the Z pole of the annihilation cross section. We predict no directional correlation of the ultrahigh-energy cosmic rays with the galactic halo. At the same time, there can be some directional correlations in the data, reflecting the distribution of matter at redshift z = z(e)+/-deltaz. Both of these features are manifest in the existing data. Our scenario is consistent with the neutrino mass reported by super Kamiokande and requires no lepton asymmetry or clustering of the background neutrinos. PMID- 11017523 TI - Lorentz and CPT tests with spin-polarized solids AB - Experiments using macroscopic samples of spin-polarized matter offer exceptional sensitivity to Lorentz and CPT violation in the electron sector. Data from existing experiments with a spin-polarized torsion pendulum provide sensitivity in this sector rivaling that of all other existing experiments and could reveal spontaneous violation of Lorentz symmetry at the Planck scale. PMID- 11017524 TI - Precision observables and electroweak theories AB - We compute the bounds from precision observables on alternative theories of electroweak symmetry breaking. We show that a cutoff as large as 3 TeV can be accommodated by the present data, without any new particles or unnatural fine tuning. PMID- 11017525 TI - Improved branching ratio measurement for the decay K(0)(L) --> &mgr;(+)&mgr;(-) AB - We report results from Experiment 871, performed at the BNL AGS, of a measurement of the branching ratio K(0)(L)-->&mgr;(+)&mgr;(-) with respect to the CP violating mode K(0)(L)-->pi(+)pi(-). This experiment detected over 6200 candidate &mgr;(+)&mgr;(-) events, a factor of 6 more than that seen in all previous measurements combined. The resulting branching ratio gamma(K(0)(L)- >&mgr;(+)&mgr;(-))/gamma(K(0)(L)-->pi(+)pi(-)) = (3. 474+/-0.057)x10(-6) leads to a branching fraction B(K(0)(L)-->&mgr;(+)&mgr;(-)) = (7.18+/-0.17)x10(-9), which is consistent with the current world average, and reduces the uncertainty in this decay mode by a factor of 3. PMID- 11017527 TI - G(E(p))/G(M(p)) ratio by polarization transfer in e-->p --> ep--> AB - The ratio of the proton's elastic electromagnetic form factors, G(E(p))/G(M(p)), was obtained by measuring P(t) and P(l), the transverse and the longitudinal recoil proton polarization, respectively. For elastic e-->p-->ep-->, G(E(p))/G(M(p)) is proportional to P(t)/P(l). Simultaneous measurement of P(t) and P(l) in a polarimeter provides good control of the systematic uncertainty. The results for the ratio G(E(p))/G(M(p)) show a systematic decrease as Q2 increases from 0.5 to 3.5 GeV2, indicating for the first time a definite difference in the spatial distribution of charge and magnetization currents in the proton. PMID- 11017526 TI - First observation of the decay B --> J/psistraight phiK AB - We present the first observation of the decay B-->J/psistraight phiK. Using 9.6x10(6) B&Bmacr; meson pairs collected with the CLEO detector, we have observed ten fully reconstructed B-->J/psistraight phiK candidates, whereas the estimated background is 0.5+/-0.2 event. We obtain a branching fraction of B(B- >J/psistraight phiK) = (8. 8(+3.5)(-3.0)[stat]+/-1.3[syst])x10(-5). This is the first observed B meson decay requiring the creation of an additional s&smacr; quark pair. PMID- 11017528 TI - Microscopic calculation of the inclusive electron scattering structure function in 16O AB - We calculate the charge form factor and the longitudinal structure function for 16O and compare with the available experimental data, up to a momentum transfer of 4 fm(-1). The ground-state correlations are generated using the coupled cluster [ exp(S)] method, together with the realistic v18 NN interaction and the Urbana IX three-nucleon interaction. Center-of-mass corrections are dealt with by adding a center-of-mass Hamiltonian to the usual internal Hamiltonian, and by means of a many-body expansion for the computation of the observables measured in the center-of-mass system. PMID- 11017529 TI - Slow spin relaxation of Rb atoms confined in glass cells filled with dense 4He gas at 1.85 K AB - At temperatures below 2.1 K, long-lived gaseous Rb atoms in glass cells have been generated with a simple method: irradiating the cells, containing 4He gas and Rb metal, with a cw laser. The obtained atomic Rb density ( approximately 10(8) cm( 3)) decreases with a 1/e time constant of about 10 s at 1.85 K. We have performed optical pumping of the Rb atoms and measured the longitudinal electronic spin relaxation time at 1.85 K as well. For processes (such as Rb-He collisions) which do not remove the atomic Rb from the vapor, this relaxation time is found to be about 60+/-15 s. PMID- 11017530 TI - Doubly vibrationally enhanced four wave mixing: the optical analog to 2D NMR. AB - We report the development of the four wave mixing vibrational analog to 2D NMR and demonstrate its spectral selectivity, sensitivity to the interactions causing mode coupling, and ability to spectrally resolve isotopic mixtures. The method discriminates against uncoupled vibrational modes and isolates the features that are associated with intra- or intermolecular interactions. PMID- 11017531 TI - Rydberg electron interferometry AB - A recent quantum mechanical study [W. Isaacs and M. A. Morrison, Phys. Rev. A 57, R9 (1998)] discovered pronounced oscillations in cross sections for near-resonant energy transfer collisions of rare-gas atoms with initially aligned Rydberg atoms. We analyze such collisions for 17d(m)-->18p(m(')) transitions in the Ca-He system semiclassically and show that the oscillations arise from a phase interference process unique to Rydberg target states. In addition to explaining the origin of these structures, this analysis explains their disappearance when the relative Ca-He velocity goes to infinity and/or the energy defect vanishes and their dependence on the initial and final magnetic quantum numbers of the transition. PMID- 11017532 TI - Nonlinear optics and quantum entanglement of ultraslow single photons AB - Two light pulses propagating with slow group velocities in a coherently prepared atomic gas exhibit dissipation-free nonlinear coupling of an unprecedented strength. This enables a single-photon pulse to coherently control or manipulate the quantum state of the other. Processes of this kind result in generation of entangled states of radiation field and open up new prospectives for quantum information processing. PMID- 11017533 TI - Annihilation of one of the coexisting attractors in a bistable system AB - A small change of one of the system parameters may not in general convert a bistable system to a monostable system. However, an external control in the form of a slow periodic parameter modulation can annihilate one of the coexisting states, and thus results in controlled monostability. The annihilation takes place because the state becomes chaotic via the period doubling route and the chaotic state undergoes boundary crisis within a small range of the control amplitude. These features are observed theoretically in two standard models, namely, Henon map and laser rate equations, and confirmed experimentally in a cavity loss modulated CO2 laser. PMID- 11017534 TI - Periodic-orbit theory of anderson localization on graphs AB - We present the first quantum system where Anderson localization is completely described within periodic-orbit theory. The model is a quantum graph analogous to an aperiodic Kronig-Penney model in one dimension. The exact expression for the probability to return to an initially localized state is computed in terms of classical trajectories. It saturates to a finite value due to localization, while the diagonal approximation decays diffusively. Our theory is based on the identification of families of isometric orbits. The coherent periodic-orbit sums within these families, and the summation over all families, are performed analytically using advanced combinatorial methods. PMID- 11017535 TI - Noncausal time response in frustrated total internal reflection? AB - Tunneling of photons in frustrated total internal reflection has been studied in the time domain with single-cycle femtosecond pulses. It is seen that both the phase and energy of the pulse travel faster than the speed of light in vacuum. Theoretical analysis of the experiments shows that the time-response function for electromagnetic waves propagating in the air gap is noncausal. However, it is found that superluminal signal propagation is not possible in this case because of the inevitable diffractive spreading of the signal beam. PMID- 11017536 TI - New effects in light scattering in disordered media and coherent backscattering cone: systems of magnetic particles AB - Single and multiple scattering of light by magnetic particles and their implications to the coherent backscattering effect are reported. Single scattering of light by small magnetic particles presents unusual features such as forward-backward asymmetry and resonance effects. In multiple scattering, this leads to a global decrease in the localization parameter kl(*), which exhibits an oscillatory dependence on the scatterer magnetic permeability. By considering magnetic scatterers following a Curie-Weiss susceptibility law, we suggest that kl(*) can be tuned by varying the temperature. PMID- 11017537 TI - Stresses in silos: comparison between theoretical models and new experiments AB - We present precise and reproducible mean pressure measurements at the bottom of a cylindrical granular column. If a constant overload is added, the pressure is linear in overload and nonmonotonic in the column height. The results are quantitatively consistent with a local, linear relation between stress components, as was recently proposed by some of us. They contradict the simplest classical (Janssen) approximation, and may rather severely test competing models. PMID- 11017538 TI - Characteristics of two-dimensional turbulence that self-organizes into vortex crystals AB - Experiments have found that freely relaxing turbulence in inviscid, incompressible two-dimensional Euler flows can self-organize into ordered structures-vortex crystals-in which a number N(c) approximately 2-20 of strong vortices form stable, rigidly rotating patterns in a low vorticity background. In this paper we show that N(c) can be roughly predicted from properties of the flows in the early stage of the turbulent relaxation. PMID- 11017539 TI - Two-dimensional evolution equation of finite-amplitude internal gravity waves in a uniformly stratified fluid AB - We derive a fully nonlinear evolution equation that can describe the two dimensional motion of finite-amplitude long internal waves in a uniformly stratified three-dimensional fluid of finite depth. The derived equation is the two-dimensional counterpart of the evolution equation obtained by Grimshaw and Yi [J. Fluid Mech. 229, 603 (1991)]. In the small-amplitude limit, our equation is reduced to the celebrated Kadomtsev-Petviashvili equation. PMID- 11017540 TI - Spontaneous fluctuations of a temperature filament in a magnetized plasma AB - This experiment illustrates the spatiotemporal pattern of the fluctuations that spontaneously develop in a magnetized temperature filament whose transverse scale is comparable to the electron skin depth. A high-frequency mode exhibits a striking spiral structure and is identified as a drift-Alfven eigenmode. A low frequency mode is found to be localized near the center of the filament. It is documented that the fluctuations significantly increase the transport of heat beyond the prediction of classical theory based on Coulomb collisions. PMID- 11017541 TI - Reconnection rate for the inhomogeneous resistivity petschek model AB - The reconnection rate for the canonical simplest case of steady-state two dimensional symmetric reconnection in an incompressible plasma is found by matching of an outer Petschek solution and an internal diffusion region solution. The reconnection rate obtained naturally incorporates both Sweet-Parker and Petschek regimes, while the latter is possible only for a strongly localized resistivity. PMID- 11017542 TI - Effect of the plasma density scale length on the direction of fast electrons in relativistic laser-solid interactions AB - The angular distribution of bremsstrahlung gamma rays produced by fast electrons accelerated in relativistic laser-solid interaction has been studied by photoneutron activation in copper. We show that the gamma-ray beam moves from the target normal to the direction of the k(laser) vector as the scale length is increased. Similar behavior is found also in 2D particle-in-cell simulations. PMID- 11017543 TI - Suppression of plasma turbulence during optimized shear configurations in JET AB - Correlation of density turbulence suppression and reduced plasma transport is observed in the internal transport barrier (ITB) region of JET tokamak discharges with optimized magnetic shear. The suppression occurs in two stages. First, low frequency turbulence and ion transport are reduced across the plasma core by a toroidal velocity shear generated by intense auxiliary heating. Then with the ITB formation, high frequency turbulence and electron transport are reduced locally within the steep pressure gradient region of the ITB. PMID- 11017544 TI - Evidence of highly collective Co diffusion in the whole stability range of Co-Zr glasses AB - Using a radiotracer technique, we have measured the isotope effect E = dln(D)/dln(m(1/2)) of Co diffusion in CoxZr1-x glasses for 0. 31 directions towards the bond center position. The activation energy for the dissociation of near-substitutional Cu is estimated to be 1.8-2.2 eV. PMID- 11017552 TI - Ab initio study of screw dislocations in Mo and ta: A new picture of plasticity in bcc transition metals AB - We report the first ab initio density-functional study of <111> screw dislocation cores in the bcc transition metals Mo and Ta. Our results suggest a new picture of bcc plasticity with symmetric and compact dislocation cores, contrary to the presently accepted picture based on continuum and interatomic potentials. Core energy scales in this new picture are in much better agreement with the Peierls energy barriers to dislocation motion suggested by experiments. PMID- 11017553 TI - Breakdown of scaling in the nonequilibrium critical dynamics of the two dimensional XY model AB - The approach to equilibrium, from a nonequilibrium initial state, in a system at its critical point is usually described by a scaling theory with a single growing length scale, xi(t) approximately t(1/z), where z is the dynamic exponent that governs the equilibrium dynamics. We show that, for the 2D XY model, the rate of approach to equilibrium depends on the initial condition. In particular, xi(t) approximately t(1/2) if no free vortices are present in the initial state, while xi(t) approximately (t/lnt)(1/2) if free vortices are present. PMID- 11017554 TI - Nagel scaling, relaxation, and universality in the kinetic ising model on an alternating isotopic chain AB - The dynamic critical exponent and the frequency and wave-vector dependent susceptibility of the kinetic Ising model on an alternating isotopic chain with Glauber dynamics are examined. The analysis provides a connection between a microscopic model and the Nagel scaling curve originally proposed to describe dielectric susceptibility measurements of several glass-forming liquids. While support is given to the hypothesis relating the Nagel scaling to multiple relaxation processes, it is also found that the scaling function may exhibit plateau regions and does not hold for all temperatures. PMID- 11017555 TI - From microscopic interactions to macroscopic laws of cluster evolution AB - We derive macroscopic governing laws of growth velocity, surface tension, mobility, critical nucleus size, and morphological evolution of clusters, from microscopic scale master equations for a prototype surface reaction system with long range adsorbate-adsorbate interactions. PMID- 11017556 TI - Attenuation of second sound in superfluid 3He-A1 AB - The attenuation of second sound (spin-entropy) wave in the superfluid A1 phase has been measured in magnetic fields up to 11 T and to sufficiently high frequency to observe the bulk attenuation proportional to the square of frequency. The measured attenuation coefficient is compared with the existing theories of hydrodynamics and dissipative coefficients. The resulting "excess" attenuation is discussed in terms of the temperature dependent spin diffusion coefficient in the superfluid. PMID- 11017557 TI - Wetting and molecular orientation of 8CB on silicon substrates AB - The wetting properties of 8CB ( 4(')-n-octyl-4-cyanobiphenyl) on silicon wafers have been studied with scanning polarization force microscopy (SPFM). Layer-by layer spreading of 8CB droplets is observed. With the help of the surface potential mapping capability of SPFM, we found that the molecular dipole of the first monolayer of 8CB is parallel to the surface. A layer of nearly vertical molecular dimers on top of the monolayer has an associated surface potential of 40 mV, which is attributed to a distortion of the dimer. The dimer distortion propagates to the subsequent smectic bilayers, producing an additional 7 mV potential increase in the second layer, 2 mV on the third, and approximately 1 mV on the fourth. PMID- 11017558 TI - Diffusion driven concerted motion of surface atoms: Ge on Ge(001) AB - The diffusion of Ge dimers on the Ge(001) surface has been studied with scanning tunneling microscopy. We have identified three different diffusion pathways for the dimers: diffusion of on-top dimers over the substrate rows, diffusion across the substrate rows, and diffusion of dimers in the trough. We report on a heretofore unknown phenomenon, namely, diffusion driven concerted motion of substrate atoms. This concerted motion is a direct consequence of the rearrangement of substrate atoms in the proximity of the trough dimer adsorption site. PMID- 11017559 TI - Control and characterization of a multistep unimolecular reaction AB - Electrons from a scanning tunneling microscope were used to break a C-H bond in a single acetylene (HCCH) molecule adsorbed on Cu(001) at 9 K. The product ethynyl (CCH) was characterized by imaging, inelastic electron tunneling spectroscopy, and variable temperature measurements of the rotation rate. Significant changes in the vibrational spectra and bonding geometry accompanied HCCH dissociation. The ethynyl was further dehydrogenated to form dicarbon (CC). Dissociation studies of the isotopes HCCD and DCCD permitted unambiguous identification of the reaction products. PMID- 11017560 TI - Inelastic neutron scattering measurements and Ab initio calculations of hydrogen in single-crystal palladium AB - Using a single-crystal sample of PdH, we have measured the inelastic form factor for exciting the proton from its ground state to various excited states as a function of crystal orientation. Because of the anharmonicity of the potential well which splits otherwise degenerate states, each inelastic form factor could be measured individually. Agreement between theory and experiment is excellent, both with regard to the transition energies and to the modulation of the intensity, giving confidence in the quantum treatment of the proton and hence in the use of the model to predict other properties of the system. PMID- 11017561 TI - Commensurability, excitation gap, and topology in quantum many-particle systems on a periodic lattice AB - In combination with Laughlin's treatment of the quantized Hall conductivity, the Lieb-Schultz-Mattis argument is extended to quantum many-particle systems (including quantum spin systems) with a conserved particle number on a periodic lattice in arbitrary dimensions. Regardless of dimensionality, interaction strength, and particle statistics (Bose or Fermi), a finite excitation gap is possible only when the particle number per unit cell of the ground state is an integer. PMID- 11017562 TI - Resolution of the scaling exponent puzzle for weakly compensated crystalline silicon and germanium metal-insulator systems AB - Using a classical theory for ionized impurity scattering, it is demonstrated that in the degenerate regime the conductivity scales as sqrt[epsilon(F)] where the Fermi energy is measured with respect to the mobility edge. The approach, a special case of alloy theory, explains the conductivity scaling exponent s = 1 / 2 observed for weakly compensated, doped crystalline Si and Ge. The results explain the breadth of scaling range and suggest how to obtain a consistent picture of the scaling of the mobility, diffusion coefficient, and Hall coefficient. PMID- 11017563 TI - Coulomb gap: how a metal film becomes an insulator AB - Electron tunneling measurements of the density of states (DOS) in ultrathin Be films reveal that a correlation gap mediates their insulating behavior. In films with sheet resistance R<5000 Omega the correlation singularity appears as the usual perturbative ln(V) zero bias anomaly (ZBA) in the DOS. As R is increased further, however, the ZBA grows and begins to dominate the DOS spectrum. This evolution continues until a nonperturbative |V| Efros-Shklovskii Coulomb gap spectrum finally emerges in the highest R films. Transport measurements of films which display this gap are well described by a universal variable range hopping law R(T) = (h/2e(2))exp(T0/T)(1/2). PMID- 11017564 TI - Angle-resonant stimulated polariton amplifier AB - We experimentally demonstrate resonant coupling between photons and excitons in microcavities which can efficiently generate enormous single-pass optical gains approaching 100. This new parametric phenomenon appears as a sharp angular resonance of the incoming pump beam, at which the moving excitonic polaritons undergo very large changes in momentum. Ultrafast stimulated scattering is clearly identified from the exponential dependence on pump intensity. This device utilizes boson amplification induced by stimulated energy relaxation. PMID- 11017565 TI - Band structure of the jahn-teller polaron from quantum monte carlo AB - A path-integral representation for the Jahn-Teller polaron (JTP) is constructed. It leads to a perturbation series that can be summed exactly by the diagrammatic quantum Monte Carlo technique. The ground-state energy, effective mass, spectrum, and density of states of the three-dimensional JTP are calculated with no systematic errors. The band structure of JTP is found to be similar to that of the Holstein polaron. The mass of JTP increases exponentially with the coupling constant. At small phonon frequencies, the spectrum of JTP is flat at large momenta, which leads to a strongly distorted density of states with a massive peak at the top of the band. PMID- 11017566 TI - Time-correlated soliton tunneling in charge and spin density waves AB - We consider a model in which an electric field induces quantum nucleation of kink antikink pairs in a pinned charge or spin density wave. Pair nucleation events, prevented by Coulomb blockade below a pair creation threshold, become correlated in time above threshold. The model provides a natural explanation for the observed (i) small density wave polarization below threshold in NbSe (3), (ii) narrow band noise, (iii) coherent oscillations, and (iv) mode-locking at high drift frequencies. PMID- 11017567 TI - Undercompensated kondo impurity with quantum critical point AB - The low-temperature properties of a magnetic impurity of spin S interacting with an electron gas via anisotropic spin exchange are studied via Bethe's ansatz. For S>1/2 the impurity is only partially compensated at T = 0, leaving an effective spin that is neither integer nor half integer. The entropy has an essential singularity at H = T = 0, and the susceptibility and the specific heat follow power laws of H and T with nonuniversal exponents, which are the consequence of a quantum critical point. The results for the generalization to an arbitrary number of channels are also reported. PMID- 11017568 TI - Enhanced fluctuations of the tunneling density of states near the bottom of a landau band measured by a local spectrometer AB - We have found that the local density of state fluctuations (LDOSF) in a disordered metal, detected using an impurity in the barrier as a spectrometer, undergo enhanced (with respect to Shubnikov-de Haas and de Haas-van Alphen effects) oscillations in strong magnetic fields, omega(c)tau>/=1. We attribute this to the dominant role of the states near the bottom of Landau bands which give the major contribution to the LDOSF and are most strongly affected by disorder. We also demonstrate that in intermediate fields the LDOSF increase with field B in accordance with the results obtained in the diffusion approximation. PMID- 11017569 TI - Probing the plateau-insulator quantum phase transition in the quantum hall regime AB - We report quantum Hall experiments on the plateau-insulator transition in a low mobility In(0.53)Ga(0.47)As/InP heterostructure. The data for the longitudinal resistance rho(xx) follow an exponential law and we extract a critical exponent kappa = 0.55+/-0. 05 which is slightly different from the established value kappa = 0. 42+/-0.04 for the plateau transitions. Upon correction for inhomogeneity effects, which cause the critical conductance sigma(*)(xx) to depend marginally on temperature, our data indicate that the plateau-plateau and plateau-insulator transitions are in the same universality class. PMID- 11017570 TI - Anisotropic vortex plasticity in the liquid state of YBa2Cu3O7: evidence for quenched c-axis vortex correlation length AB - A generalized methodology, based on anisotropic angular dependent magnetoresistance measurements, is presented to deconvolute the contributions to the vortex activation energy (i.e., plastic and pinning energies) in the vortex liquid state of high temperature superconductors. Experimental evidence is given for the appearance of a partially entangled liquid vortex state in YBa 2Cu 3O (7) when random quenched and correlated disorders compete, as in twinned melt textured YBa 2Cu 3O (7) quasisingle crystals with Y 2BaCuO (5) precipitates. The hallmark of this new phase is a quench of the c-axis vortex correla-tion length. PMID- 11017571 TI - Observation of out-of-phase bilayer plasmons in YBa(2)Cu(3)O(7-delta) AB - The temperature dependence of the c-axis optical conductivity sigma(omega) of optimally and overdoped YBa2Cu3Ox ( x = 6.93 and 7) is reported in the far- (FIR) and midinfrared (MIR) range. Below T(c) we observe a transfer of spectral weight from the FIR not only to the condensate at omega = 0, but also to a new peak in the MIR. This peak is naturally explained as a transverse out-of-phase bilayer plasmon by a model for sigma(omega) which takes the layered crystal structure into account. With decreasing doping the plasmon shifts to lower frequencies and can be identified with the surprising and so far not understood FIR feature reported in underdoped bilayer cuprates. PMID- 11017572 TI - Accurate prediction of large antiferromagnetic interactions in high- T(c) HgBa2Ca(n-1)Cu(n)O(2n+2+delta) ( n = 2,3) superconductor parent compounds AB - The in-plane nearest-neighbor Heisenberg magnetic coupling constant, J, of La2CuO4, Nd2CuO4, Sr2CuO2Cl2, YBa2Cu3O6, and undoped HgBa(2)Ca(n 1)Cu(n)O(2n+2+delta) ( n = 1,2,3) is calculated from accurate ab initio configuration interaction calculations. For the first four compounds, the theoretical J values are in quantitative agreement with experiment. For the Hg based compounds the predicted values are -135 meV ( n = 1) and approximately -160 meV ( n = 2,3), the latter being much larger than in previous cases and, for n = 3, increasing with pressure. Nevertheless, the physics governing J in all these layered cuprates appears to be the same. Moreover, calculations suggest a possible relationship between J and T(c). PMID- 11017573 TI - Imaging of a vortex lattice transition in YNi2B2C by scanning tunneling spectroscopy AB - The vortex lattices in YNi2B2C under the magnetic fields H up to 3 T applied along both the a and the c axes have been studied by scanning tunneling spectroscopy at 4.2 K. The vortex lattice transition has been found to occur in different manners for H parallela and H parallelc; in H parallela a slightly distorted hexagonal vortex lattice has been found to transform to a nearly square one above 1.0 T with increasing H, while in H parallelc the transition occurs at a much lower field around 0.1 T. The unconventional steep increase of the quasiparticle density of states outside the vortex core has also been found well below H(c2). PMID- 11017574 TI - Pressure dependence of the irreversibility line in Bi2Sr2CaCu2O(8+delta): role of anisotropy in flux-line formation AB - One of the important problems of high-temperature superconductivity is to understand and ultimately to control fluxoid motion. Here we present data on the pressure dependence of the irreversibility line measured up to 2.5 GPa. We observe that the application of pressure changes the interplanar coupling by decreasing the c-axis length, without significantly disturbing the intraplanar superconductivity. Our results directly show the relationship between lattice spacing and the irreversibility line in Bi(2)Sr(2)CaCu(2)O(8+delta), and demonstrate the potential for a dramatic reduction in the flux motion. PMID- 11017575 TI - Photoemission quasiparticle spectra of Sr2RuO4 AB - Multiband quasiparticle calculations based on perturbation theory and dynamical mean-field methods show that the creation of a photoemission hole state in Sr2RuO4 is associated with a highly anisotropic self-energy. Since the narrow Ru derived d(xz,yz) bands are more strongly distorted by Coulomb correlations than the wide d(xy) band, charge is partially transferred from d(xz,yz) to d(xy), thereby shifting the d(xy) Van Hove singularity close to the Fermi level. PMID- 11017576 TI - Spin-triplet superconductivity in Sr2RuO4 probed by andreev reflection AB - The superconducting gap function of Sr2RuO4 was investigated by means of quasiparticle reflection and transmission at the normal conductor-superconductor interface of Sr2RuO4-Pt point contacts. We found two distinctly different types of dV/dI vs V spectra either with a double-minimum structure or with a zero-bias conductance anomaly. Both types of spectra are expected in the limit of high and low transparency, respectively, of the interface barrier between a normal metal and a spin-triplet superconductor. Together with the temperature dependence of the spectra this result strongly supports a spin-triplet superconducting order parameter for Sr2RuO4. PMID- 11017577 TI - Phase separation in supersolids AB - We study quantum phase transitions in the ground state of the two dimensional hard-core boson Hubbard Hamiltonian. Recent work on this and related models has suggested "supersolid" phases with simultaneous diagonal and off-diagonal long range order. We show numerically that, contrary to the generally held belief, the most commonly discussed "checkerboard" supersolid is thermodynamically unstable. Furthermore, this supersolid cannot be stabilized by next-near-neighbor interaction. We obtain the correct phase diagram using the Maxwell construction. We demonstrate that the "striped" supersolid is thermodynamically stable and is separated from the superfluid phase by a continuous phase transition. PMID- 11017578 TI - Cooperative jahn-teller coupling in the manganites AB - The cooperative Jahn-Teller coupling between the Mn e(g) electrons and the oxygen octahedral distortions in LaMnO3 is studied using ab initio density-functional calculations and tight-binding models. The linear and quadratic vibronic coupling parameters are calculated using density-functional methods. It is shown that the cooperative Jahn-Teller coupling, primarily due to the interoctahedral electron hopping (band structure term), leads to the ordering of the octahedral distortion and simultaneously to orbital ordering. The coupling results in a two-minima adiabatic potential surface in the solid, instead of the three-minima "Mexican hat" surface for the isolated octahedron. PMID- 11017579 TI - Negatively charged quantum well polaritons in a GaAs/AlAs microcavity: An analog of atoms in a cavity AB - The negatively charged exciton (X-) is observed to strongly couple with the microcavity- (MC-)confined photons in a GaAs quantum well containing a two dimensional electron gas with 00, consistent with the scattering at the nodes of the d-wave gap. A superlinear sigma(q)(H) marks the normal state above T(c). PMID- 11017626 TI - Quasiparticles in the superconducting state of Bi(2)Sr(2)CaCu(2)O(8+delta) AB - Recent improvements in momentum resolution lead to qualitatively new angle resolved photoemission spectroscopy results on the spectra of Bi(2)Sr(2)CaCu(2)O(8+delta) (Bi2212) along the (pi,pi) direction, where there is a node in the superconducting gap. We now see the intrinsic line shape, which indicates the presence of true quasiparticles at all Fermi momenta in the superconducting state, and lack thereof in the normal state. The region of momentum space probed here is relevant for charge transport, motivating a comparison of our results to conductivity measurements by infrared reflectivity. PMID- 11017627 TI - Do columnar defects produce bulk pinning? AB - From magneto-optical imaging performed on heavy-ion-irradiated YBa(2)Cu(3)O(7 delta) single crystals, it is found that at fields and temperatures where strong single vortex pinning by individual irradiation-induced amorphous columnar defects is to be expected, vortex motion is limited by the nucleation of vortex kinks at the specimen surface. In the material bulk, vortex motion occurs through (easy) kink sliding. Depinning in the bulk determines the screening current only at fields comparable to or larger than the matching field, at which the majority of vortices is not trapped by an ion track. PMID- 11017628 TI - Metastability and paramagnetism in superconducting mesoscopic disks AB - A projected order parameter is used to calculate not only local minima of the Ginzburg-Landau energy functional but also saddle points or energy barriers responsible for the metastabilities observed in superconducting mesoscopic disks [A. K. Geim et al., Nature (London) 396, 144 (1998)]. We calculate the local minima magnetization and find the energetic instability points between vortex configurations with different vorticity. We also find that, for any vorticity, the supercurrent can reverse its flow direction on decreasing the magnetic field before one vortex can escape. PMID- 11017629 TI - Theoretical model for the superconducting and magnetically ordered borocarbides AB - We present a theory of superconductivity in the presence of a general magnetic structure in a form suitable for the description of complex magnetic phases encountered in borocarbides. The theory, complemented with some details of the band structure and with the magnetic phase diagram, may explain the nearly reentrant behavior and the anisotropy of the upper critical field of HoNi2B2C. The onset of the helical magnetic order depresses superconductivity via the reduction of the interaction between phonons and electrons caused by the formation of magnetic Bloch states. At mean field level, no additional suppression of superconductivity is introduced by the incommensurability of the helical phase. PMID- 11017630 TI - Andreev reflection in strong magnetic fields AB - We have studied the interplay of Andreev reflection and cyclotron motion of quasiparticles at a superconductor-normal-metal interface with a strong magnetic field applied parallel to the interface. Bound states are formed due to the confinement introduced by both the external magnetic field and the superconducting gap. These bound states are a coherent superposition of electron and hole edge excitations similar to those realized in finite quantum-Hall samples. We find the energy spectrum for these Andreev edge states and calculate transport properties. PMID- 11017631 TI - Exact demonstration of magnetization plateaus and first-order dimer-Neel phase transitions in a modified shastry-sutherland model for SrCu2(BO3)(2) AB - We study a generalized Shastry-Sutherland model for the material SrCu2(BO3)(2). Along a line in the parameter space, we show rigorously that the model has a first-order phase transition between dimerized and Neel-ordered ground states. Furthermore, when a magnetic field is applied in the dimerized phase, magnetization plateaus develop at commensurate values of the magnetization. We also discuss various aspects of the phase diagram and properties of this model away from this exactly soluble line, which include gap-closing continuous transitions between dimerized and magnetically ordered phases. PMID- 11017632 TI - Origin of biquadratic exchange in Fe/Si/Fe AB - The thickness and temperature dependences of the interlayer exchange coupling in well-defined molecular beam epitaxy-grown Fe/Si/Fe sandwich structures have been studied. The biquadratic coupling shows a strong temperature dependence in contrast to the bilinear coupling. Both depend exponentially on thickness. These observations can be well understood in the framework of Slonczewski's loose spins model [J. Appl. Phys. 73, 5957 (1993)]. No bilinear contribution of the loose spins to the coupling was observed. PMID- 11017633 TI - Domain-wall induced coupling between ferromagnetic layers AB - The remanent magnetization of a hard ferromagnetic CoPtCr layer is progressively decreased by repeated switching of a neighboring soft magnetic layer. We show that this effect depends strongly on the thickness of the CoPtCr layer and the spacing between the hard and soft layers. We propose a model that accounts for these results: An interlayer magnetostatic coupling is induced by large stray fields from domain walls that form within the soft layer during its magnetization reversal. PMID- 11017634 TI - Polarized raman study of aligned multiwalled carbon nanotubes AB - Polarized Raman spectra of high purity aligned arrays of multiwalled carbon nanotubes, prepared on silica substrates from the thermal decomposition of a ferrocene-xylene mixture, show a strong dependence of the graphitelike G band and the disorder-induced D band on the polarization geometry employed in the experiments. The experimental G-band intensity exhibits a minimum at straight theta(m) = 55 degrees in the VV configuration, in good agreement with theoretical predictions of a characteristic minimum at 54.7 degrees for A(1g) modes in single wall nanotubes, where straight theta(m) denotes the angle between the polarization direction and the nanotube axis. PMID- 11017635 TI - Statistical mechanics of systems with heterogeneous agents: minority games AB - We study analytically a simple game theoretical model of heterogeneous interacting agents. We show that the stationary state of the system is described by the ground state of a disordered spin model which is exactly solvable within the simple replica symmetric ansatz. Such a stationary state differs from the Nash equilibrium where each agent maximizes her own utility. The latter turns out to be characterized by a replica symmetry broken structure. Numerical results fully agree with our analytical findings. PMID- 11017636 TI - Random walks in the space of conformations of toy proteins. AB - Monte Carlo dynamics of the lattice toy protein of 48 monomers is interpreted as a random walk in an abstract (discrete) space of conformations. To test the geometry of this space, we examine the return probability P(T), which is the probability to find the polymer in the native state after T Monte Carlo steps, provided that it starts from the native state at the initial moment. Comparing computational data with the theoretical expressions for P(T) for random walks in a variety of different spaces, we show that conformation spaces of polymer loops may have nontrivial dimensions and exhibit negative curvature characteristics of Lobachevskii (hyperbolic) geometry. PMID- 11017637 TI - Factorial moments analyses show a characteristic length scale in DNA sequences. AB - A unique feature of most of the DNA sequences, found through the factorial moments analysis, is the existence of a characteristic length scale around which the density distribution is nearly Poissonian. Above this point, the DNA sequences, irrespective of their intron contents, show long range correlations with a significant deviation from the Gaussian statistics, while, below this point, the DNA statistics are essentially Gaussian. The famous DNA walk representation is also shown to be a special case of the present analysis. PMID- 11017638 TI - Partition function zeros and finite size scaling of helix-coil transitions in a polypeptide. AB - We report on multicanonical simulations of the helix-coil transition of a polypeptide. The nature of this transition was studied by calculating partition function zeros and the finite size scaling of various quantities. New estimates for critical exponents are presented. PMID- 11017639 TI - Fluid-solid phase separation in hard-sphere mixtures is unrelated to bond percolation PMID- 11017640 TI - Buhot replies: PMID- 11017641 TI - Comment on "Zero temperature phases of the electron gas" PMID- 11017642 TI - Ortiz et al. reply: PMID- 11017643 TI - Nonclassical states: An observable criterion AB - An observable criterion is derived that allows one to distinguish nonclassical states of the harmonic oscillator from those having a classical counterpart. A quantum state is shown to have no classical counterpart if and only if the characteristic functions of the quadrature distributions or the s-parametrized phase-space distributions exhibit a slower decay than for the ground state of the oscillator. This renders it possible to experimentally check the failure of the P function to be a probability measure. PMID- 11017644 TI - Stable monopole and dyon solutions in the einstein-yang-mills theory in asymptotically anti-de sitter space AB - A continuum of new monopole and dyon solutions in the Einstein-Yang-Mills theory in asymptotically anti-de Sitter space are found. They are regular everywhere and specified by their mass and their non-Abelian electric and magnetic charges. A class of monopole solutions which have no node in non-Abelian magnetic fields is shown to be stable against spherically symmetric linear perturbations. PMID- 11017645 TI - Observation of the cabibbo-suppressed decay xi(+)(c) --> pK(-)pi(+) AB - We report the first observation of the Cabibbo-suppressed charm baryon decay Xi(+)(c)-->pK(-)pi(+). We observe 150+/-22+/-5 events for the signal. The data were accumulated using the SELEX spectrometer during the 1996-1997 fixed target run at Fermilab, chiefly from a 600 GeV/c Sigma(-) beam. The branching fractions of the decay relative to the Cabibbo-favored Xi(+)(c)-->Sigma+K-pi(+) and Xi(+)(c)-->Xi(-)pi(+)pi(+) are measured to be B(Xi(+)(c)-->pK(-)pi(+))/B(Xi(+)(c) ->Sigma+K-pi(+)) = 0.22+/-0. 06+/-0.03 and B(Xi(+)(c)-->pK(-)pi(+))/B(Xi(+)(c)- >Xi(-)pi(+)pi(+)) = 0.20+/-0.04+/-0.02, respectively. PMID- 11017647 TI - Partial dynamical symmetry in a fermion system AB - The relevance of the partial dynamical symmetry concept for an interacting fermion system is demonstrated. Hamiltonians with partial SU(3) symmetry are presented in the framework of the symplectic shell model of nuclei and shown to be closely related to the quadrupole-quadrupole interaction. Implications are discussed for the deformed light nucleus 20Ne. PMID- 11017646 TI - Nuclear and coulomb interaction in 8B breakup at sub-coulomb energies AB - The angular distribution for the breakup of 8B-->7Be+p on a 58Ni target has been measured at an incident energy of 25.75 MeV. The data are inconsistent with first order theories but are remarkably well described by calculations including higher order effects. The comparison with theory illustrates the importance of the inclusion of the exotic proton halo structure of 8B in accounting for the data. PMID- 11017648 TI - Quantum energy flow in atomic ions moving in magnetic fields AB - Using a combination of semiclassical and recently developed wave packet propagation techniques we find the quantum self-ionization process of highly excited ions moving in magnetic fields which has its origin in the energy transfer from the center of mass to the electronic motion. It obeys a time scale orders of magnitude larger than the corresponding classical process. Importantly a quantum coherence phenomenon leading to the intermittent behavior of the ionization signal is found and analyzed. Universal properties of the ionization process are established. PMID- 11017649 TI - Photoassociation spectroscopy of cold He(2 3S) atoms AB - We observe vibrational states by photoassociation spectroscopy of cold He(2 3S) atoms. Photoassociation resonances are detected as peaks in the Penning ionization rate over a frequency range of 20 GHz below the atomic 2 3S1-2 3P2 transition frequency. We have observed three vibrational series, of which two can be identified. A possible mechanism to explain the observed increase of the Penning ionization rate is discussed. PMID- 11017650 TI - Complete numerical solution of electron-hydrogen model collision problem above the ionization threshold AB - Benchmark results are presented for electrons colliding with hydrogen atoms in the S-wave (Temkin-Poet) model collision problem, which neglects angular momentum. Complete results (elastic, inelastic, and ionization), accurate to 1%, are obtained by numerically integrating Schrodinger's equation subject to correct asymptotic boundary conditions. This marks the first time direct matching to asymptotic boundary conditions has been shown to yield convergent ionization amplitudes for a Coulomb three-body problem. Results are presented for impact energies of 54.4 and 40.8 eV, where comparison with other theories is available. PMID- 11017651 TI - Dynamical casimir effect at finite temperature AB - Thermal effects on the creation of particles under the influence of time dependent boundary conditions are investigated. The dominant temperature correction to the energy radiated by a moving mirror is derived by means of response theory. For a resonantly vibrating cavity the thermal effect on the number of created photons is obtained nonperturbatively. Finite temperatures can enhance the pure vacuum effect by several orders of magnitude. The relevance of finite-temperature effects for the experimental verification of the dynamical Casimir effect is addressed. PMID- 11017652 TI - Correlations in the adiabatic response of chaotic systems AB - Adiabatic variation of the parameters of a chaotic system results in a fluctuating reaction force. The quantum analog of a classical dissipative force, proportional to the time integral of the force-force correlation function, vanishes. We study this quantum-classical crossover for random matrix models. For the Gaussian unitary ensemble the crossover is found to take place on the Heisenberg time scale and the finite time integral practically vanishes for longer times. For the Gaussian orthogonal case, there is no such time scale and the integral falls off inversely proportional to time. PMID- 11017653 TI - Prediction of spatiotemporal time series based on reconstructed local states AB - Spatiotemporal time series are analyzed and predicted using reconstructed local states. As numerical examples the evolution of a Kuramoto-Sivashinsky equation and a coupled map lattice are predicted from previously sampled data. PMID- 11017654 TI - Finite flux solutions of the quantum boltzmann equation and semiconductor lasers AB - We propose and illustrate in the context of the semiconductor laser that, in nonequilibrium fermionic systems with sources and sinks, the family of finite flux stationary solutions of the quantum Boltzmann equation is central and more important then the zero flux Fermi-Dirac spectrum. We present the quantum analog of the finite flux Kolmogorov spectra which are central to understanding nonequilibrium classical systems such as high Reynolds number hydrodynamics and the wave turbulence encountered in water waves, plasmas, and optics. In particular, we show how semiconductor laser efficiency can be improved by maximizing the flux of carriers (electrons and holes) towards the lasing frequencies. PMID- 11017655 TI - Intrinsic bistability of luminescence and stimulated emission in Yb- and Tm-doped glass AB - Intrinsic bistability and hysteresis have been observed in the photoluminescence of bulk glass doped with Yb3+ and Tm3+ impurities at room temperature. Multiple instabilities also occur in channel waveguide lasers in this material. Consequently, local field effects exhibit more degrees of freedom than expected and have important implications for compact optical amplifiers and oscillators. PMID- 11017656 TI - Cantor set fractals from solitons AB - We show how a nonlinear system that supports solitons can be driven to generate exact (regular) Cantor set fractals. As an example, we use numerical simulations to demonstrate the formation of Cantor set fractals by temporal optical solitons. This fractal formation occurs in a cascade of nonlinear optical fibers through the dynamical evolution from a single input soliton. PMID- 11017657 TI - Spatiotemporally localized multidimensional solitons in self-induced transparency media AB - "Light bullets" are multidimensional solitons which are localized in both space and time. We show that such solitons exist in two- and three-dimensional self induced transparency media and that they are fully stable. Our approximate analytical calculation, backed and verified by direct numerical simulations, yields the multidimensional generalization of the one-dimensional sine-Gordon soliton. PMID- 11017658 TI - Propagating pattern selection and causality reconsidered AB - Pattern selection, occurring when a nonuniform state of a nonlinear dissipative system propagates into an initially unstable, homogeneous basic state is reconsidered by application of the causality principle. In particular, the nonlinear marginal stability criterion that determines the selection of a nonlinear front solution is replaced by an exact general necessary condition that has never been considered before. The demonstration is based on the causal signaling problem derived in the context of plasma physics. PMID- 11017659 TI - Astrophysical jets as exact plasma equilibria AB - Two families of exact global solutions to the equations of plasma equilibrium are derived. The solutions model astrophysical jets and solar prominences and provide counterexamples to Parker's hypothesis. PMID- 11017660 TI - Negative ion density fronts during ignition and extinction of plasmas in electronegative gases AB - Negative ion density fronts have been shown to occur in electronegative steady state plasmas with hot electrons. In this Letter, we report theoretical and numerical results on the spatiotemporal evolution of negative ion density fronts during plasma ignition and extinction (afterglow). During plasma ignition, the negative ion fronts are analogous to hydrodynamic shocks. This is not the case during plasma extinction where, although negative ions diffuse freely in the plasma core, the negative ion front propagates towards the chamber walls with a nearly constant velocity. PMID- 11017661 TI - Impurity-induced suppression of core turbulence and transport in the DIII-D tokamak AB - Turbulence is significantly reduced in a tokamak plasma as a result of neon seeding of an L-mode discharge. Correspondingly, confinement is improved and cross-field ion thermal transport reduced. Fully saturated turbulence in the range 0.10.35. These observations are consistent with a reduction in the calculated linear growth rate for k( perpendicular)rho(s)>0.5 and an increase in the measured ExB flow shearing rate. PMID- 11017662 TI - Internal atomic distortion and layer roughness of epitaxial SiC thin films studied by short wavelength X-Ray diffraction AB - Epitaxial silicon carbide thin films grown on a silicon substrate were examined by short wavelength ( 0.71 A from a molybdenum anode) x-ray diffraction to study their atomic distortion and internal layer roughness. Up to 5 order Bragg peaks along (100) were measured and crystallographic structure factors were obtained. Electron density distributions along the surface normal were reconstructed via Fourier transform. In comparing to the ideal situation, it was found that, due to the lattice mismatch, the silicon atoms are no longer fixed on the same atomic plane, thus the corresponding electron distribution maxima were broadened. Also, the maxima for carbon atoms are flattened, indicating the randomized locations. PMID- 11017663 TI - Photoalignment of liquid crystals by liquid crystals AB - We observed light-induced alignment of the nematic liquid crystal, 4, 4(')-n pentylcyanobiphenyl (5CB), on a fused quartz. Irradiation of the adsorbed layer with polarized ultraviolet light produced homogeneous alignment in a 5CB-filled liquid crystal cell with the easy orientation axis perpendicular to the polarization direction. The measured anchoring energy increases with illumination up to 10(-4) erg/cm (2). Phase retardation and pretilt measurements confirmed near homogeneous alignment of the LC in a cell, while the polarization dependence of second harmonic generation suggested a near normal alignment of the adsorbed layer. We believe that light-induced rearrangement or phototransformation of the adsorbed layer causes the observed phenomenon. PMID- 11017664 TI - Director precession and nonlinear waves in nematic liquid crystals under elliptic shear AB - Elliptic shear applied to a homeotropically oriented nematic above the electric bend Freedericks transition (FT) generates slow precession of the director. The character of the accompanying nonlinear waves changes from diffusive phase waves to dispersive ones exhibiting spirals and spatiotemporal chaos as the FT is approached from above. An exact solution of the flow alignment equations captures the observed precession and predicts its reversal for non-flow-aligning materials. The FT transforms into a Hopf bifurcation opening the way to understand the wave phenomena. PMID- 11017665 TI - Magnetic field dependent tunneling in glasses AB - We report on experiments giving evidence for quantum effects of electromagnetic flux in barium alumosilicate glass. In contrast to expectation, below 100 mK the dielectric response becomes sensitive to magnetic fields. The experimental findings include both lifting of the dielectric saturation by weak magnetic fields and oscillations of the dielectric response in the low temperature resonant regime. As the origin of these effects we suggest that the magnetic induction field violates the time reversal invariance leading to a flux periodicity in the energy levels of tunneling systems. At low temperatures, this effect is strongly enhanced by the interaction between tunneling systems and thus becomes measurable. PMID- 11017666 TI - Charged local defects in extended systems AB - The conventional approach to treat charged defects in extended systems in first principles calculations is via the supercell approximation using a neutralizing jellium charge. I explicitly show that errors in the resulting electrostatic potential surface are comparable to a band gap energy in semiconductors, for cell sizes typically used in simulations. I present a method for eliminating divergence of the Coulomb potential in charged supercell calculations that correctly treats the electrostatic potential in the local vicinity of a charged defect, via a mixed boundary condition approach. PMID- 11017667 TI - Scaling and noise in slow combustion of paper AB - We present results of high resolution experiments on kinetic roughening of slow combustion fronts in paper, focusing on short length and time scales. Using three different grades of paper, we find that the combustion fronts show apparent spatial and temporal multiscaling at short scales. The scaling exponents decrease as a function of the order of the corresponding correlation functions. The noise affecting the fronts reveals short range temporal and spatial correlations, and non-Gaussian noise amplitudes. Our results imply that the overall behavior of slow combustion fronts cannot be explained by standard theories of kinetic roughening. PMID- 11017668 TI - Dynamic scaling of ultrasonic damping near the nematic-smectic- A transition of TBBA AB - We report a detailed study of the ultrasonic damping near the nematic-smectic- A phase transition of terephtal-bis- p-p(')-butylaniline (TBBA). Two mechanisms contribute to the damping. One is isotropic; it is associated with the critical fluctuations, and leads to a scaling behavior of the 3D- XY type which is observed up to 15 degrees C from the transition, even though the transition is first order. The other is anisotropic; it appears only in the smectic- A phase, and is associated with the relaxation of the order-parameter modulus. PMID- 11017669 TI - Orientational ordering of N2O molecules adsorbed on graphite (0001): A novel commensurate pinwheel structure AB - The structure and orientational ordering of N2O molecules physisorbed on graphite (0001) is investigated applying x-ray, neutron, and low-energy electron diffraction techniques. Combining the results of the three techniques, we find that N2O forms a highly ordered, hexagonal, commensurate (sqrt[21]xsqrt[21])R10.89 degrees phase. The unit cell contains seven molecules which are arranged in a seven-sublattice pinwheel structure, unexpected for linear molecules on a hexagonal lattice. Potential energy calculations corroborate these results. PMID- 11017671 TI - Polarizability of KC60: evidence for potassium skating on the C60 surface AB - We present the first measurement of the polarizability and the permanent dipole moment of isolated KC60 molecules by molecular beam deflection technique. We have obtained a value of 2506+/-250 A(3) for the polarizability at room temperature. The addition of a potassium atom enhances by more than a factor of 20 the polarizability of a pure C60 molecule. This very high polarizability and the lack of observed permanent dipole show that the apparent polarizability of KC60 is induced by the free skating of the potassium atom on the C60 surface, resulting in a statistical orientation of the dipole. The results are interpreted with a simple model similar to the Langevin theory for paramagnetic systems. PMID- 11017670 TI - Strain evolution in coherent Ge/Si islands AB - Strain evolution of coherent Ge islands on Si(001) is measured using a newly developed transmission electron microscopy technique based on two-beam dark-field strain imaging. The strain measurements show that a metastable Ge island shape is involved in the shape transition between pyramids and domes; this shape is more readily observed for growth at 550 than 600 degrees C because of the slower rate at which islands cross the kinetic barrier between shapes. The strain relaxation changes discontinuously between pyramids and domes, indicating that the underlying shape transition is first order. PMID- 11017672 TI - Anomalous shape and decay of islands on Au(110) AB - Scanning tunneling microscopy on roughened Au(110) reveals that the equilibrium shape of islands and pits on this surface is almondlike: each island contains two smoothly curved steps joined at two sharp corners. This shape has recently been predicted and finds its origin in the missing-row reconstruction of its fcc (110) surfaces (Au, Pt, etc.). We use the corner angles and the island shapes to determine the step energies. In addition we find that during the decay of an island on the Au(110) surface the shape changes and that the disappearance of the island involves the splitting of the layer below the island into two disconnected regions. The shape change has a dramatic influence on the decay rate of the islands. PMID- 11017673 TI - Mesoscopic sensitivity of speckles in disordered nonlinear media to changes of the scattering potential AB - We show that the sensitivity of wave speckle patterns in disordered nonlinear media to changes of scattering potential increases with sample size. For large sizes the sensitivity diverges, which implies that for a given coherent wave incident on a sample there are multiple solutions for the spatial distribution of the wave density. The number of solutions increases exponentially with the sample size. PMID- 11017674 TI - Fractional quantum conductance in carbon nanotubes AB - Using a scattering technique based on a parametrized linear combination of atomic orbitals Hamiltonian, we calculate the ballistic quantum conductance of multiwall carbon nanotubes. We find that interwall interactions not only block some of the quantum conductance channels, but also redistribute the current nonuniformly over individual tubes across the structure. Our results provide a natural explanation for the unexpected integer and noninteger conductance values reported for multiwall nanotubes by Stefan Frank et al. [Stefan Frank et al., Science 280, 1744 (1998)]. PMID- 11017675 TI - Spectral function of composites from reflectivity measurements AB - We demonstrate a method of calculating the spectral function of a composite from measured reflectivity data. To solve this inverse problem it is necessary for the reflectivity data to be taken through a strong, high Q, resonance. By analyzing the reststrahlen region of different fill fraction KCl-diamond composites at three different temperatures, we find accurate spectral functions that are independent of temperature with the low temperature data giving the best resolution. These spectral functions are then used to successfully predict the optical response of RbCl-diamond composites. PMID- 11017676 TI - Nematic phase of the two-dimensional electron gas in a magnetic field AB - The two-dimensional electron gas (2DEG) in moderate magnetic fields in ultraclean AlAs-GaAs heterojunctions exhibits transport anomalies suggestive of a compressible anisotropic metallic state. Using scaling arguments and Monte Carlo simulations, we develop an order parameter theory of an electron nematic phase. The observed temperature dependence of the resistivity anisotropy behaves like the orientational order parameter if the transition to the nematic state occurs at a finite temperature T(c) approximately 65 mK, and is slightly rounded by a small background microscopic anisotropy. We propose a light scattering experiment to measure the critical susceptibility. PMID- 11017677 TI - Double quantum dots as detectors of high-frequency quantum noise in mesoscopic conductors AB - We propose a measurement setup for detecting quantum noise over a wide frequency range using inelastic transitions in a tunable two-level system as a detector. The frequency-resolving detector consists of a double quantum dot which is capacitively coupled to the leads of a nearby mesoscopic conductor. The inelastic current through the double quantum dot is calculated in response to equilibrium and nonequilibrium current fluctuations in the nearby conductor, including zero point fluctuations at very low temperatures. As a specific example, the fluctuations across a quantum point contact are discussed. PMID- 11017678 TI - Large isotope effect on the pseudogap in the high-temperature superconductor HoBa2Cu4O8 AB - The oxygen isotope effect on the relaxation rate of crystal-field excitations in the slightly underdoped high-temperature superconductor HoBa2Cu4O8 has been investigated by means of inelastic neutron scattering. For the 16O compound there is clear evidence for the opening of an electronic gap in the normal state at T(*) approximately 170 K far above T(c) = 79 K. Upon oxygen isotope substitution ( 16O vs 18O) T(c) decreases marginally to 78.5 K, whereas T(*) is shifted to about 220 K. This huge isotope shift observed for T(*) which is absent in NMR and NQR experiments suggests that the mechanism leading to an isotope effect on the pseudogap has to involve a time scale in the range 10(-8)>>tau>10(-13) s. PMID- 11017679 TI - Vortices freeze like window glass: the vortex molasses scenario AB - We overview several recent experimental and numerical observations, which are at odds with the vortex glass theory of the freezing of disordered vortex matter. To reinvestigate the issue, we performed numerical simulations of the overdamped London-Langevin model, and use finite size scaling to analyze the data. Upon approaching the transition the initial vortex-glass-type criticality is arrested at some crossover temperature. Below this temperature the time scales continue growing very quickly, consistent with the Vogel-Fulcher form, while the spatial correlation length xi stops exhibiting any observable divergence. We call this mode of freezing the vortex molasses scenario. PMID- 11017680 TI - Anomalous magnetotransport properties of R2Mo2O7 near the magnetic phase boundary AB - The magnetic and transport properties of pyrochlore R2Mo2O7 have been studied with variation of the rare earth ( R). The change of the mean ionic radius of R, which induces change of the lattice structure, determines the magnetic ground state (ferromagnetic or spin glass), and the magnetic phase boundary is correlated with the metal-insulator crossover. Furthermore, we found enhanced magnetoresistance and unusual residual anomalous Hall effect at low temperatures near the phase boundary, which can be attributed to the coexistence of both phases. PMID- 11017681 TI - Strong surface state effects in nonlinear magneto-optical response of Ni(110). AB - Spectroscopic magnetization-induced optical second harmonic generation (MSHG) measurements from a clean Ni(110) surface reveal strong resonance effects near 2.7 eV that can be attributed to the presence of an empty surface state. The good agreement with model calculations shows the potential of MSHG to probe spin polarized interface band structures. PMID- 11017682 TI - Femtosecond coherent dynamics of the fermi-edge singularity and exciton hybrid. AB - We study theoretically the coherent nonlinear optical response of doped quantum wells with several subbands. When the Fermi energy approaches the exciton level of an upper subband, the absorption spectrum acquires a characteristic double peak shape originating from the interference between the Fermi-edge singularity and the exciton resonance. We demonstrate that, for off-resonant pump excitation, the pump-probe spectrum undergoes a striking transformation, with a time dependent exchange of oscillator strength between the Fermi-edge singularity and exciton peaks. This effect originates from the many-body electron-hole correlations which determine the dynamical response of the Fermi sea. PMID- 11017683 TI - Many-body renormalization of semiconductor quantum wire excitons: absorption, gain, binding, and unbinding. AB - We consider theoretically the formation and stability of quasi-one-dimensional many-body excitons in GaAs quantum wire structures under external photoexcitation conditions by solving the dynamically screened Bethe-Salpeter equation for realistic Coulomb interaction. In agreement with several recent experimental findings the calculated excitonic peak shows weak carrier-density dependence up to (and even above) the Mott transition density, nc approximately 3 x 10(5) cm( 1). Above nc we find considerable optical gain demonstrating compellingly the possibility of a one-dimensional quantum wire laser operation. PMID- 11017684 TI - Limits for entanglement measures. AB - The basic principle of entanglement processing says that entanglement cannot increase under local operations and classical communication. Based on this principle, we show that any entanglement measure E suitable for the regime of a high number of identically prepared entangled pairs satisfies ED < or = E < or = EF, where ED and EF are the entanglement of distillation and formation, respectively. Moreover, we exhibit a theorem establishing a very general form of bounds for distillable entanglement. PMID- 11017685 TI - Viscous sintering phenomena in liquid-liquid dispersions. AB - We present experimental evidence for viscous sintering phenomena in a gel formed by highly viscous emulsion droplets. When a rupturing agent is added to the initially stable emulsion, a gel forms, which further contracts by preserving the geometry of the container. The initial stages of densification (up to 60%) follow very well the "cylindrical model" for viscous sintering, but deviate at the final stages of densification. The observed inverse dependence of the contraction rate on viscosity is consistent with the viscous sintering theory. PMID- 11017686 TI - Observation of chaotic dynamics in dilute sheared aqueous solutions of CTAT. AB - The nonlinear flow behavior of a viscoelastic gel formed due to entangled, cylindrical micelles in aqueous solutions of the surfactant cetyl trimethylammonium tosilate (CTAT) has been studied. On subjecting the system to a step shear rate lying above a certain value, the shear and normal stresses show interesting time dependent behavior. The analysis of the measured time series shows the existence of a finite correlation dimension and a positive Lyapunov exponent, unambiguously implying that the dynamics can be described by that of a dynamical system with a strange attractor whose dimension increases with the increase in shear rate. PMID- 11017687 TI - Glassy transition in a disordered model for the RNA secondary structure. AB - We numerically study a disordered model for the RNA secondary structure and we find that it undergoes a phase transition, with a breaking of the replica symmetry in the low temperature region (as in spin glasses). Our results are based on the exact evaluation of the partition function. PMID- 11017688 TI - Cryptographical properties of Ising spin systems. AB - The relation between Ising spin systems and public-key cryptography is investigated using methods of statistical physics. The insight gained from the analysis is used for devising a matrix-based cryptosystem whereby the ciphertext comprises products of the original message bits; these are selected by employing two predetermined randomly constructed sparse matrices. The ciphertext is decrypted using methods of belief propagation. The analyzed properties of the suggested cryptosystem show robustness against various attacks and competitive performance to modern cryptographical methods. PMID- 11017689 TI - Comment on "Physical picture for light emission in scanning tunneling microscopy". PMID- 11017691 TI - Comment on "Precision neutron interferometric search for evidence of nuclear quantum entanglement in liquid H2O-D2O Mixtures". PMID- 11017693 TI - Comment on "Adatom formation on the Ni(110) surface". PMID- 11017695 TI - Comment on "Complementarity enforced by random classical phase kicks". PMID- 11017697 TI - Comment on "Carrier density collapse and colossal magnetoresistance in doped manganites". PMID- 11017699 TI - Comment on "Density matrix renormalization group study of the Haldane phase in random one-dimensional antiferromagnets". PMID- 11017701 TI - Relative merits of optical biosensors based on flow-cell and cuvette designs. PMID- 11017702 TI - Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: comparison of endpoint and real-time methods. AB - Four quantitative reverse transcription-PCR (RT-PCR) methods were compared to evaluate the time course of mRNA formation and decay. Mouse fibroblasts (NIH 3T3) transfected with the human beta-globin open reading frame/c-myc 3'-untranslated region chimeric gene under control of the c-fos promoter (fos-glo-myc) were used for serum-inducible transcription. The amount of fos-glo-myc mRNA, relative to beta-actin, was measured by quantitative, RT-PCR at various times following the addition of serum to serum-starved fibroblasts transfected with the chimeric gene. Both endpoint (band densitometry and probe hybridization) and real-time (SYBR green and TaqMan) PCR methods were used to assay the identical cDNA. The real-time methods produced a 4- to 5-log dynamic range of amplification, while the dynamic range of the endpoint assays was 1-log. The real-time and probe hybridization assays produced a comparable level of sensitivity that was considerably greater than band densitometry. The coefficient of variation from 22 replicate PCR reactions was 14.2 and 24.0% for the SYBR green and TaqMan detection, respectively, and 44.9 and 45.1% for the band densitometry and probe hybridization assays, respectively. The rank order for the values of r(2) obtained from the linear regression of the first-order mRNA decay plots was SYBR green > TaqMan > probe hybridization > band densitometry. Real-time PCR is more precise and displays a greater dynamic range than endpoint PCR. Among the real time methods, SYBR green and TaqMan assays produced comparable dynamic range and sensitivity while SYBR green detection was more precise and produced a more linear decay plot than TaqMan detection. PMID- 11017703 TI - Electrospray mass spectra of three proprietary detergents. AB - We have determined the major ingredients of the commercially available reagents M PER, Y-PER, and B-PER from Pierce Chemical Co. using electrospray mass spectrometry. These three proprietary reagents have been widely used in the biochemical community as cell membrane dissolving tools during the initial step of protein purification. However, the identity and mechanism of these reagents remained unknown. In this paper, we identified these reagents as 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate, N-tetradecyl-N,N-dimethyl-3 ammonio-1-propanesulfonate, and n-octyl-beta-d-thioglucopyranoside, respectively. In addition, we wish to stress here the increasing importance of the role of electrospray mass spectrometry in the analysis of such proprietary biological preparations which are increasingly finding their way into the biochemical literature. PMID- 11017704 TI - Differential use of Alcian blue and toluidine blue dyes for the quantification and isolation of anionic glycoconjugates from cell cultures: application to proteoglycans and a high-molecular-weight glycoprotein synthesized by articular chondrocytes. AB - Alcian blue and toluidine blue dyes form complexes with anionic glycoconjugates (AG) such as proteoglycans (PG) and glycosaminoglycans (GAG). However, the Alcian blue-AG complexes do not readily dissociate, while the toluidine blue-AG complexes do so in salt solutions. This differential dissociation of the dye-AG complexes has been utilized in the analysis and isolation of radiolabeled AG elaborated by articular chondrocyte cultures incubated with the radiolabeled precursors of AG. For the rapid quantification of newly synthesized (35)S-labeled PG, small replicate aliquots of the radiolabeled culture media were applied directly to cellulose acetate strips, stained with Alcian blue and the stained immobilized radiolabeled PG was quantified by liquid scintillation counting. Comparison of anionic glycoconjugates quantified in the culture media employing toluidine blue and Alcian blue staining on cellulose acetate trips gave similar results. Staining on cellulose acetate strips using these two dyes is particularly suited for the simultaneous processing of large numbers of samples, as illustrated by the screening of the effects of biological materials and drugs on AG synthesis, in cultures labeled with [(35)S]-sulfate and [(3)H]-glucosamine. The Alcian blue and toluidine blue precipitation methods yielded similar results for the total AG recovered from the media of TGF-beta-stimulated chondrocytes. Electrophoretic analysis of toluidine blue- and Alcian blue-precipitated AG followed by autoradiography and Alcian blue staining in combination with silver nitrate demonstrated that both dyes yielded similar pattern of bands on gels. However, some AG from Alcian blue precipitate did not enter the gel, suggesting incomplete dissociation of Alcian blue-AG complex. The application of the toluidine blue precipitation method, in combination with enzymatic digestion of the GAG chains of the PGs, is illustrated by the isolation of a non-PG high molecular-weight AG, as well as the PGs from the media of chondrocyte cultures stimulated by TGF-beta. PMID- 11017705 TI - A standardized spectrophotometric assay of endoglycanase activities using dyed, amorphous polysaccharides. AB - This is a new technique to assay virtually any endoglycanase activity where enough polysaccharide material is available to allow for production of the amorphous, dyed beads used as substrates. It allows for a direct comparison of endoglycanase activities between laboratories since dyed beads from at least the most common polysaccharides such as cellulose, xylan, mannan, and chitin are now under development and will soon be commercially available; cellulose beads already are. It is a very sensitive technique and enzyme activities can be measured using a nonsophisticated spectrophotometer. PMID- 11017706 TI - Selective and sensitive biosensor for theophylline based on xanthine oxidase electrode. AB - Milk and microbial xanthine oxidases (XOs) were used for the construction of amperometric enzyme electrodes. Substrate specificity differences of these enzymes were studied. Of the two enzymes, only the microbial XO was found to oxidize theophylline, but not theobromine and caffeine. The substrate specificity of microbial XO was affected by pH, where the optimum for xanthine was 5.5, while for theophylline it was in the range from 6.5 to 8.5. The theophylline biosensor showed a low detection limit of 2 x 10(-7) M and signal linearity up to 5 x 10( 5) M. The sensitivity of the microbial XO electrode to theophylline could be selectively eliminated by immersion in alkaline phosphate solution, thus allowing for the construction of a blank electrode for differential measurements. The feasibility of this approach has been demonstrated by the determination of free (unbound) and total theophylline in blood samples. The biosensor exhibited good operational (>6 h) and shelf (>3 months) stability when trehalose was used as a stabilizer of the biocatalytic layer. PMID- 11017707 TI - A highly sensitive high-performance liquid chromatography-- fluorometric method for the assay of peptidylarginine deiminase activity. AB - The activity of peptidylarginine deiminase (PAD) has generally been assayed by a colorimetric method using N-benzoyl-L-arginine ethyl ester (BAEE) and N-benzoyl-L arginine (Bz-L-Arg) as the substrates. The widespread occurrence of citrulline and urea in tissues makes use of this method difficult, especially for small samples. We developed a highly sensitive high-performance liquid chromatography method with N-dansyl-glycyl-L-arginine as the substrate. This method was sensitive enough to determine previously undetectable activity of PAD in HL-60 cells. Two types of PAD (HL-60 cell and brain PAD) could be distinguished by differential competition, using either BAEE or Bz-L-Arg as a preferential substrate in the assay. These data indicate that the present method is applicable to many tissues. PMID- 11017708 TI - How to measure and analyze tryptophan fluorescence in membranes properly, and why bother? AB - Tryptophan fluorescence is a powerful tool for studying protein structure and function, especially membrane-active proteins and peptides. It is arguably the most frequently used tool for examining the interactions of proteins and peptides with vesicular unilamellar model membranes. However, high light scattering associated with vesicular membrane systems presents special challenges. Because of their reduced light scattering compared to large unilamellar vesicles (LUV), small unilamellar vesicles (SUV) produced by sonication are widely used membrane models. Unfortunately, SUV, unlike LUV, are metastable and consequently unsuitable for equilibrium thermodynamic measurements. We present simple and easily implemented experimental procedures for the accurate determination of tryptophan (Trp) fluorescence in either LUV or SUV. Specifically, we show that Trp spectra can be obtained in the presence of up to 6 mM LUV that are virtually identical to spectra obtained in buffer alone, which obviates the use of SUV. We show how the widths and peak positions of such spectra can be used to evaluate the heterogeneity of the membrane conformation and penetration of peptides. Finally, we show how to use a reference fluorophore for the correction of intensity measurements so that the energetics of peptide partitioning into membranes can be accurately determined. PMID- 11017709 TI - Separation and quantitation of phospholipids and lysophospholipids by high performance liquid chromatography. AB - We describe a comprehensive approach to the separation, quantitation, and characterization of phospholipids and lysophospholipids present in complex biological samples. The central feature is a normal-phase HPLC separation of individual phospholipid and lysophospholipid classes. In this single chromatographic step, phospholipids and lysophospholipids are separated and recovered for quantitation by organic phosphate assay and characterization by acyl-group composition. Recovery of phospholipids and lysophospholipids from HPLC averages 80-90%. Isolated phospholipid and lysophospholipid fractions are available for separation of individual molecular species by second-dimension reverse-phase HPLC and characterization of individual molecular species by mass spectrometry. PMID- 11017710 TI - Assay of glutathione in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection. AB - Glutathione (GSH) in individual mouse peritoneal macrophages was determined by capillary zone electrophoresis with electrochemical end-column amperometric detection at a gold/mercury amalgam microelectrode. A capillary of 20 microm inner diameter was suitable for determination of GSH in an individual macrophage with a good signal-to-noise ratio. Individual macrophages could be drawn into the capillary with the aid of a inverted microscope. Lysing cells was studied in different buffer solutions. 0.01 mol/liter NaOH was selected to lyse macrophages. In this method, the usual calibration curve of GSH could not be used for the quantification of GSH in individual macrophages. It was found that standard GSH injected after analyzing each cell could be served as external standard. The whole cell injection and the lack of necessity of a derivatization reaction lead to more accurate and precise results. The average amount of GSH in an individual mouse peritoneal macrophage is 5.8 fmol, which is consistent with the literature value. PMID- 11017711 TI - Introduction of C3 exoenzyme into cultured endothelium by lipofectamine. PMID- 11017712 TI - A colorimetric method for determining distearoylphosphatidylethanolamine- polyethylene glycol 2000 in blood suspension. PMID- 11017713 TI - Quantitative polymerase chain reaction to estimate the number of amplified esterase genes in insecticide-resistant mosquitoes. PMID- 11017714 TI - Quantification of phosphatidylinositol 3,4,5-trisphosphate by liposome lysis assay with specific monoclonal antibodies. PMID- 11017715 TI - A new Western blotting method using polymer immunocomplexes: detection of Tsc1 and Tsc2 expression in various cultured cell lines. PMID- 11017716 TI - Clean Western blots of membrane proteins after yeast heterologous expression following a shortened version of the method of Perini et al. PMID- 11017717 TI - Simple and efficient cloning of small polymerase chain reaction-generated DNA products. PMID- 11017718 TI - Editorial PMID- 11017719 TI - The development of working memory: exploring the complementarity of two models. AB - The aim of the paper was to further explore the complementarity of the working memory models postulated by Pascual-Leone and Baddeley. Five-, six-, eight-, and nine-year-old children were assessed on two working memory tasks that have frequently been used within the respective streams of research: the Mr. Peanut task and the Corsi blocks task. Results indicated a developmental increase in spatial short-term memory for both tasks. Concurrent spatial suppression reduced performance on the two tasks in all four age groups. By contrast, articulatory suppression interfered with recall only on the Mr. Peanut task, and in only the older children. The two models were shown to make their own specific contribution to the interpretation of the data, attesting to their complementarity. Pascual Leone's theory offered a clear explanation of the results concerning the central aspects of working memory, that is, the stepwise age-related increase in performance, whereas Baddeley's model provided a convincing account of the findings regarding the peripheral phonological and visuo-spatial components, that is, the effects of articulatory and spatial suppression. PMID- 11017720 TI - The development of working memory: further note on the comparability of two models of working memory. AB - This paper is a set of reflections on Kemps, De Rammelaere, and Desmet's article (2000, this issue), in which the two models by Baddeley and Pascual-Leone are compared. First, some of the similarities and differences between the two models which we identified in a 1994 paper (de Ribaupierre & Bailleux, 1994) are briefly summarized and reexamined in the light of more recent work. Second, we debate the issue of whether each model makes a specific contribution to the explanation of some of Kemps et al.'s results, that is, of whether they can be considered to be complementary. Third, we argue for the necessity of theoretical task analyses, in view of the divergent results obtained in the two tasks used (the Corsi and the Peanut tasks), notably different developmental profiles, and an overall higher level of performance in the Corsi task. Finally, we briefly summarize a very similar study in which we also used Mr. Peanut with concurrent tasks in children and in young adults and in which we obtained rather different results. By comparing the experimental procedures used in the two studies, we contribute some exploratory hypotheses, while raising issues that can easily be generalized to other visuo-spatial working memory tasks. PMID- 11017721 TI - Development of working memory: should the Pascual-Leone and the Baddeley and Hitch models be merged? AB - The data presented by Kemps, De Rammelaere, and Desmet (2000, this issue) appear to have some aspects that fit most readily into our own model (Baddeley & Hitch, 1974), while others appear to support that of Pascual-Leone (1970). We accept that our initial model said little about development and was better able to account for relatively simple memory-based tasks than more complex cognitive activities. More recent elaborations of the model are, however, able to throw new light on the processes underlying cognitive development, offering a better account than that provided by existing neo-Piagetian interpretations. Meanwhile, the addition of a fourth component to the model, namely the episodic buffer, offers a way of dealing with more complex cognitive activities. Given the major differences between our own model and that of Pascual-Leone in basic assumptions, and in theoretical style, we suggest that any attempt to combine the two would be premature. PMID- 11017722 TI - Reflections on working memory: are the two models complementary? AB - By means of the theoretical modeling of data from Kemps, De Rammelaere, and Desmet (2000, this issue), the working memory theory of Baddeley and the theory of constructive operators of Pascual-Leone are contrasted and compared. It is concluded that although the theory of constructive operators is complementary with working memory theory (for it explains developmental and individual differences that working memory theory cannot explain), the converse is not true; the theory of constructive operators explains all the data without need of working memory theory. PMID- 11017723 TI - The effects of spatial movement components precues on the execution of rapid aiming in children aged 7, 9, and 11. AB - The aim of the present study was twofold: first, to investigate the effects of spatial precues on the execution of rapid aiming in children aged 7, 9, and 11 and second, to provide a kinematic support to the investigation of the role of precues in aiming tasks performed under temporal constraints. Four precuing conditions were used, where participants received: (a) no precue of any type, (b) advance information on direction, (c) advance information on amplitude, and (d) complete information on the forthcoming movement. Our results showed that precuing the spatial dimensions of movement shortens reaction times, that such shortening is a function of the number of precued parameters, and that spatial precues modify the kinematics of the children's rapid aiming movements. Peak velocity increased with direction and/or amplitude, suggesting that precues play a significant role in motor preparation. Moreover, the accuracy results indicate that direction precuing induces a proactive directional regulation. Finally, direction and amplitude appear to be independently specified in children. PMID- 11017724 TI - Pk-fit: a pharmacokinetic/pharmacodynamic and statistical data analysis software. AB - This paper presents a new software, Pk-fit, to fit nonlinear models to kinetic and dynamic data. Directly connected to the spreadsheet, a statistical software component manager is available. In the data manager, Pk-fit includes the noncompartmental analysis module, the compartmental analysis module, the nonlinear kinetic process module, the drug absorption module, the pharmacodynamic data modeling module, the simultaneous fitting module, and the user-defined library module. In this paper, we present a detailed comparison of the kinetic analysis using Pk-fit and common software packages, PCNONLIN, MODFIT, MKMODEL, NONMEM, and SIPHAR, based on the textbook published by Gabrielsson in 1992, "Compilation of Analyzed Data Sets for Pharmacokinetic Software Evaluation." The comparison of Pk-fit with the reference softwares revealed that the parameters and their dispersion found with Pk-fit are consistent with the ones estimated with the other programs. In conclusion, Pk-fit constituted a valid tool for pharmacokinetic/pharmacodynamic data analysis. PMID- 11017725 TI - Automated sleep stage scoring using hybrid rule- and case-based reasoning. AB - We propose an automated method for sleep stage scoring using hybrid rule- and case-based reasoning. The system first performs rule-based sleep stage scoring, according to the Rechtschaffen and Kale's sleep-scoring rule (1968), and then supplements the scoring with case-based reasoning. This method comprises signal processing unit, rule-based scoring unit, and case-based scoring unit. We applied this methodology to three recordings of normal sleep and three recordings of obstructive sleep apnea (OSA). Average agreement rate in normal recordings was 87.5% and case-based scoring enhanced the agreement rate by 5.6%. This architecture showed several advantages over the other analytical approaches in sleep scoring: high performance on sleep disordered recordings, the explanation facility, and the learning ability. The results suggest that combination of rule based reasoning and case-based reasoning is promising for an automated sleep scoring and it is also considered to be a good model of the cognitive scoring process. PMID- 11017726 TI - On the parallelization of linkmap from the LINKAGE/FASTLINK package. AB - Genetic linkage calculations can be time consuming, even on a fast computer. The ability to collect large family pedigrees has increased the magnitude of linkage computations. Sequential genetic algorithms have many successful applications in very different domains, but they have a main drawback in their utilization. Evaluations are very time-consuming, e.g., a pedigree consisting of 55 nodes takes about 70 min on a DEC-Alpha processor and about 270 min on a 166 MHz Pentium for certain likelihood calculations. This time increases exponentially with the increase in the size of the pedigree. In order to solve these shortcomings and to study new models of higher efficiency and efficacy, parallel platforms are being used for genetic programs. LINKAGE is a software package for performing genetic likelihood calculations; FASTLINK is an improved, faster version of it. This paper provides a parallel implementation of the "Linkmap" program (one of the four programs in LINKAGE/FASTLINK) for a heterogeneous environment, using a static and a dynamic strategy for task allocation. It was found that the increased performance by the dynamic strategy was close to the estimated maximum speed up. PMID- 11017727 TI - Artificial neural networks compared to factor analysis for low-dimensional classification of high-dimensional body fat topography data of healthy and diabetic subjects. AB - Subcutaneous adipose tissue thickness was measured in 590 healthy subjects at 15 specific body sites by means of the new optical device, lipometer, providing a high-dimensional and partly highly intercorrelated set of data, which had been analyzed by factor analysis previously. N-2-N back-propagation neural networks are able to perform low-dimensional display of high-dimensional data as a special application. We report about the performance of such a 15-2-15 network and compare its results with the output of factor analysis. As test data for verification, measurement values on women with proven diabetes mellitus type II (NIDDM) are used. Surprisingly our 15-2-15 neural network is able to reproduce the classification pattern resulting from factor analysis very precisely. After extracting the network weights the classification of new subjects is even more simple with the neural network as compared with factor analysis. In addition, the network weights are able to cluster highly correlated body sites nicely to different groups, corresponding to different regions of the human body. Thus, the analysis of these weights provides additional information about the structure of the data. Therefore, N-2-N networks seem to be a good alternative method for analyzing high-dimensional data with strong intercorrelation. PMID- 11017728 TI - Examination of the Hydrotropic Effect of Sodium p-Toluenesulfonate on a Nonionic Surfactant (C(12)E(6)) Solution. AB - Although hydrotropy is extensively used in industry, the molecular mechanism of hydrotropic solubilization has not been completely elucidated yet. In this paper the interaction between a nonionic surfactant (ethoxylated fatty alcohol containing between five and six oxyethylenic units) and sodium p-toluene sulfonate is examined. Surface tension measurements confirm that the hydrotropic effect occurs at a concentration in which the hydrotropes self-associate. Photon correlation spectroscopy studies show that for this concentration of hydrotropes a drastic reduction in the surfactant micellar radius occurs. Furthermore the luminescence of the hydrotrope used as a fluorescence probe indicates that at low concentrations p-toluene sulfonate dissolves in the surfactant micelles but beyond the minimum concentration for hydrotropic solubilization the hydrotrope is present in the aqueous phase. These results suggest that the hydrotropic effect is related to alterations in the water structure induced by the hydrotrope molecules and to the presence of hydrotrope aggregates that furnish an appropriate niche for the surfactant amphiphile. Copyright 2000 Academic Press. PMID- 11017729 TI - Effect of Insoluble Surfactants on Drainage and Rupture of a Film between Drops Interacting under a Constant Force. AB - The deformation, drainage, and rupture of an axisymmetrical film between colliding drops in the presence of insoluble surfactants under the influence of van der Waals forces is studied numerically at small capillary and Reynolds numbers and small surfactant concentrations. Constant-force collisions of Newtonian drops in another Newtonian fluid are considered. The mathematical model is based on the lubrication equations in the gap between drops and the creeping flow approximation of Navier-Stokes equations in the drops, coupled with velocity and stress boundary conditions at the interfaces. A nonuniform surfactant concentration on the interfaces, governed by a convection-diffusion equation, leads to a gradient of the interfacial tension which in turn leads to additional tangential stress on the interfaces (Marangoni effects). The mathematical problem is solved by a finite-difference method on a nonuniform mesh at the interfaces and a boundary-integral method in the drops. The whole range of the dispersed to continuous-phase viscosity ratios is investigated for a range of values of the dimensionless surfactant concentration, Peclet number, and dimensionless Hamaker constant (covering both "nose" and "rim" rupture). In the limit of the large Peclet number and the small dimensionless Hamaker constant (characteristic of drops in the millimeter size range) a fair approximation to the results is provided by a simple expression for the critical surfactant concentration, drainage being virtually uninfluenced by the surfactant for concentrations below the critical surfactant concentration and corresponding to that for immobile interfaces for concentrations above it. Copyright 2000 Academic Press. PMID- 11017730 TI - Fluidification of Concentrated Aqueous Colloidal Silica Suspensions by Adsorption of Low-Molecular-Weight Poly(ethylene oxide). AB - This paper deals with the effect of different low-molecular-weight poly(ethylene oxide)s on the rheology of concentrated aqueous colloidal silica suspensions (volume fraction >0.2) with the aim of obtaining well-dispersed media. Results are correlated with the physico-chemical characteristics of the systems that govern the ranges of the various operating interactions, i.e., mainly surface coverage, molecular weight of the polymer, and ionic strength of the medium. Optimization of the fluidification occurs to be strongly linked to these parameters. An unexpected effect of free polymer bulk concentration leads to improved fluidification when the characteristic lengths of the system are correctly adjusted; it has been interpreted in the frame of recent theories. Copyright 2000 Academic Press. PMID- 11017731 TI - Kinetics of Cream Formation by the Mechanism of Consolidation in Flocculating Emulsions. AB - In this work we propose a theoretical description of the process of creaming of batch emulsions when a sharp boundary exists between a clear serum phase and the sedimenting drops. The creaming is represented as a continuous consolidation of partially aggregated network. The treatment reproduces correctly the trend for gradually decreasing rate of sedimentation as time goes on. The theoretical results are compared quantitatively with experimental measurements of the creaming rate. Oil-in-water systems, stabilized by proteins (beta-lactoglobulin (BLG), bovine serum albumin, and mixtures BLG+beta-casein) were investigated. Faster creaming is attributed to larger size of the sedimenting objects (flocs of emulsion droplets). In systems obeying the creaming mechanism with sharp boundary (SB) the flocs are smaller when the protein concentration is higher. This supports the hypothesis for the stabilizing role of the excess amount of protein (forming lumps and multilayers on the interface). Theoretical analysis demonstrates that the formation of flocs by gravitational coagulation is a much faster process than the consolidation of the cream. Hence, the dispersions first flocculate and then cream. With increasing beta-casein content in mixtures BLG+beta-casein the emulsions depart from the SB-type behavior and are characterized by the presence of small nonflocculated droplets, which do not sediment (the serum is turbid and the boundary with the concentrated dispersion is diffuse, DB behavior). This is connected with hindered flocculation, perhaps due to beta-casein's augmented ability to prevent droplet aggregation. Copyright 2000 Academic Press. PMID- 11017732 TI - Contribution to the Determination of Kinetic Parameters of the Sol-Gel Transformation by Rheological Measurements. AB - The system tetraethoxysilane(TEOS)-water-ethanol has been studied by rheological measurements. Different molar ratios of TEOS : water (1 : 4, 1 : 10, and 1 : 20) are studied at different temperatures (30, 40, and 50 degrees C). The dynamic viscosity (rotating mode) at a constant shear rate (100 s(-1)) and the elastic and viscous moduli (oscillating mode) at a constant frequency (1 Hz) are determined. The viscosity-time curves are evaluated by application of a nucleation and particle growth model. Good agreement between experiments and theory is observed. The model allows the determination of the complex rate constant of silica precipitation. The temperature-dependent measurements gave the possibility to determine the apparent energy of activation by common methods. The results are in agreement with data from the literature. The gel time defined as intersection point of elastic and plastic moduli and its dependence on temperature are evaluated by the Smoluchowski model. The energy of activation for the coagulation was determined and found to be in the correct order of magnitude. Copyright 2000 Academic Press. PMID- 11017733 TI - Assembly of Alternated Multivalent Ion/Polyelectrolyte Layers on Colloidal Particles. Stability of the Multilayers and Encapsulation of Macromolecules into Polyelectrolyte Capsules. AB - Alternating adsorption of multivalent ions and oppositely charged polyelectrolytes on colloid particles has been investigated. Multilayer films composed of Tb(3+)/polysterene sulfonate (PSS) and 4-pyrene sulfate/polyallylamine (PAH) were successfully assembled on polysterene sulfonate (PS) and melamine formaldehyde (MF) latex particles. The amount of assembled material was estimated by fluorescence and the linear growth of the film versus the number of layers was demonstrated. These multilayers are not stable and can be decomposed by salt and temperature. Dissolution of MF particles leads to formation of hollow capsules consisting of multivalent ion/polyelectrolyte multilayers. Comparative analysis of the capsules was done by confocal and scanning force microscopy. Complex hollow spheres consisting of Tb(3+)/PSS or 4 PS/PAH as an inner shell and stable PSS/PAH as an outer shell were produced. Due to selective permeability of the outer shell after degradation of the inner shell the multivalent ions are released out of the capsule while the polyelectrolytes fill the capsule interior. This is indicative of swelling of the capsule by osmotic pressure. The filled capsules were studied by confocal and scanning electron microscopy. Possibilities of encapsulating macromolecules in defined amounts per capsule are discussed. Copyright 2000 Academic Press. PMID- 11017734 TI - Adsorption of Amphiphilic Dimers at Surfaces. AB - Adsorption of amphiphilic dimers is analyzed in the framework of density functional Ono-Kondo theory. There are three configurations for dimers absorbed at a surface: one parallel to the surface and two perpendicular to the surface (AB and BA, with A or B touching the surface, respectively). Densities of molecules in each configuration are calculated from density functional theory and compared to Monte Carlo simulation data. There is good agreement between theory and simulations. It is shown that the parallel configuration is preferred over the perpendicular configuration, except when there are very strong asymmetries in intermolecular forces. In most cases, the parallel configuration is even preferred over the combination of the two perpendicular configurations. Copyright 2000 Academic Press. PMID- 11017735 TI - Molecular Interactions in Mixed Monolayers of Octadecanoic Acid and Three Related Amphiphiles. AB - Binary mixed monolayers of octadecanoic acid and three related amphiphilic compounds (octadecanamide, octadecylamine, octadecylurea) have been investigated at the air/water interface by surface pressure-area (Pi-A) isotherms and their resistances to water evaporation (r). In addition, the excess free energies of mixing (DeltaG(E)) were calculated using the Goodrich method. Both the ln r vs x and DeltaG(E) vs x plots exhibit marked deviations from linearity, indicating a high degree of miscibility and nonideal behavior of the components in the mixed films. For all of these binary systems the excess free energies of mixing have been found to be minimum for a certain composition corresponding almost to a maximum in evaporation resistances. Weak interactions were detected in octadecanoic acid/octadecanamide monolayers, whereas significant condensation effects were observed in 1 : 1 mixed films containing octadecanoic acid and octadecylamine. This is attributed to an acid-base equilibrium followed by the formation of a well-ordered arrangement of COO(-) and NH(3)(+) head groups bound to each other by electrostatic forces. The unusual polymorphism of octadecylurea monolayers could be influenced by adding small amounts of octadecanoic acid. The formation of the low-temperature phase (beta-phase) is completely suppressed, if the acid content exceeds 8 mol%. The octadecanoic acid seems to induce the formation of the high-temperature phase (alpha-phase), which is characterized by a vertical orientation of the hydrocarbon chains. Copyright 2000 Academic Press. PMID- 11017736 TI - Adsorption Isotherms of Cetylpyridinium Chloride with Iron III Salts at Air/Water and Silica/Water Interfaces. AB - The interaction of iron III salts and cetylpyridinium chloride (CPC) has been studied at the air/water and silica/water interfaces. The surface tension of cetylpyridinium chloride has been determined in aqueous solutions in the presence of iron III chloride and iron III nitrate at two constant pH values, namely, 3.5 and 1.2. It is shown that the surface tension of the cationic surfactant depends upon the ionic strength of the solution through the pH adjustment in the presence of the former salt but not in the presence of the latter. The effect of iron III nitrate on the surface tension of CPC is similar to that of potassium nitrate, indicating that the iron III various-hydrolyzed species do not interfere with the composition of the air/water interface. The competitive adsorption of iron III nitrate salt and the cationic surfactant at a silica/water interface was next investigated. The adsorption isotherms were determined at pH 3.5. It is shown that although the iron III ions, which were added to the silica dispersion in the presence of the cetylpyridinium ions, were strongly bound to the anionic surface sites, the surfactant ions are not salted out in the solution but remain in close vicinity of the silica surface. Conversely as the cationic surfactant is added first to the silica dispersion in the presence of the adsorbed iron III ions, the metal ions and the surfactant ions are both coadsorbed onto the silica surface. It is suggested that iron III hydrolyzed or free cations and the cationic surfactant molecules may not compete for the same adsorption sites onto the silica surface. Copyright 2000 Academic Press. PMID- 11017737 TI - Multilayer Charged Structures in Nonpolar Dielectric Liquids. AB - Theory of the multilayer charged structures adjacent to an electrode surface in nonpolar dielectric liquids with low conductivity under the action of an electric field is developed. Structures of this kind have been revealed by the probe measurements of the field strength in the vicinity of the flat electrode in hydrocarbon liquids. Copyright 2000 Academic Press. PMID- 11017738 TI - Nitrogen Adsorption Studies of PAN-Based Activated Carbon Fibers Prepared by Different Activation Methods. AB - Polyacrylonitrile (PAN)-based activated carbon fibers (ACFs) prepared by various activation methods were characterized using low-temperature nitrogen adsorption over a wide relative pressure from 10(-6) to 1. Nitrogen adsorption is a standard tool for determination of porous structure parameters. In the present work, we carried out extensive adsorption studies of a series of PAN-ACFs activated by different methods. It was shown that the high-resolution alpha(S) plot provided valuable information about structural properties of samples under study. The pore size distributions of samples under study were calculated by employing the regularization method according to density functional theory. By these analyses, the pore development and the dominant pores of samples prepared by different methods can be clearly observed. Moreover, the adsorption measurement could provide profound insight into the structural heterogeneity of the ACFs. Copyright 2000 Academic Press. PMID- 11017739 TI - Characterization of Titania/Silica Gel by Means of Low-Pressure Nitrogen Adsorption. AB - Adsorbents synthesized by grafting of titania onto mesoporous silica gel surfaces at different temperatures were studied by means of nitrogen adsorption-desorption and water desorption. The pore size distribution f(R(p)) of titania/silica gel depends on the titania concentration (C(TiO(2))) and the temperature of titania synthesis. Nonuniformity of TiO(2) phase is maximal at a low C(TiO(2)) value (3.2 wt.% anatase deposited at 473 K), and two peaks of the fractal dimension distribution f(D) are observed at such a concentration of titania, but at larger C(TiO(2)) values, only one f(D) peak is seen. More ordered filling of pores and adsorption sites by nitrogen, reflecting in the shape of adsorption energy distributions f(E) at different pressures of adsorbate, is observed for adsorbent with titania (rutile+anatase) grafted on silica gel at a higher temperature (673 K). Copyright 2000 Academic Press. PMID- 11017740 TI - Activity and Conformation of Yeast Alcohol Dehydrogenase (YADH) Entrapped in Reverse Micelles. AB - Yeast alcohol dehydrogenase (YADH) solubilized in reverse micelles of aerosol OT (i.e., AOT or sodium bis (2-ethyl hexyl) sulfosuccinate) in isooctane has been shown to be catalytically more active than that in aqueous buffer under optimum conditions of pH, temperature, and water content in reverse micelles. Studies of the secondary structure conformational changes of the enzyme in reverse micelles have been made from circular dichroism spectroscopy. It has been seen that the conformation of YADH in reverse micelles is extremely sensitive to pH, temperature, and water content. A comparison has been made between the catalytic activity of the enzyme and the alpha-helix content in the conformation and it has been observed that the enzyme is most active at the maximum alpha-helix content. While the beta-sheet content in the conformation of the entrapped enzyme was found to be dependent on the enzyme-micelle interface interaction, the alpha helix and random coil conformations are governed by the degree of entrapment and the extent of rigidity provided by the micelle core to the enzyme structure. Copyright 2000 Academic Press. PMID- 11017741 TI - A Phenomenon of the Change in Particle Drift Velocity Direction in High-Field Electrophoresis. AB - The sign-alternating electric field in rectangular impulses has been used to eliminate linear electrophoresis and to study nonlinear electrophoresis in water at strong fields. We found out that the particle drift velocity could change its direction with the growth of the strength of the field. This new phenomenon has obtained its explanation in the framework of the Debye-Huckel theory of strong electrolyte where we consider a particle as a "heavy" effective ion and take into account all relevant nonlinear effects, grounding ourselves on basic physics. With the help of our theory we have succeeded in good fitting of our experimental data on black oil and Al(2)O(3) particles in distilled water using reasonable values for the basic parameters such as the Debye screening length. Copyright 2000 Academic Press. PMID- 11017742 TI - Influence of Sodium Periodate and Tyrosinase on Binding of Alginate to Adlayers of Mytilus edulis Foot Protein 1. AB - Mytilus edulis foot protein 1 (Mefp-1) is the most well-characterized component of this sea mussel's adhesive plaque. The plaque is a condensed, heterogeneous mixture consisting of a large proportion of cross-linked biopolymers that bonds the mussel to a chosen mooring. Mefp-1 is densely populated with lysine and L-3,4 dihyroxyphenylalanine (L-dopa) residues incorporated into a repeating amino acid sequence motif. It has been proposed that one plaque cross-linking reaction is the nucleophilic addition of the epsilon-amino groups of the lysine residues into the oxidized catechol (o-diphenol) functionality (quinone) of the L-dopa residues. In order to determine if this reaction occurs in adlayers of Mefp-1, a previously developed assay for epsilon-amino groups was applied. Adlayers of Mefp 1 were exposed to an oxidant, either the enzyme, mushroom tyrosinase, or sodium periodate. Binding of alginate to adlayers was used to probe for accessibility of epsilon-amino groups. It was found that lysine residues lose the ability to bind alginate after exposure to sodium periodate, but that this loss is not clearly due to a reaction with L-dopa residues. There is a slight decrease of binding of alginate to adlayers of Mefp-1 exposed to either active or thermally deactivated mushroom tyrosinase, probably due to the obstruction of binding sites by bound enzyme. Adsorption kinetics of mushroom tyrosinase onto adlayers of Mefp-1 for active and thermally inactivated enzyme were nearly identical. Attenuated total reflection Fourier transform infrared spectroscopy was used to characterize these interactions at a germanium (Ge) interface. Copyright 2000 Academic Press. PMID- 11017743 TI - Transport of Water and Ions Through a Clay Membrane. AB - A model for hindered transport of water and ions is used to predict transient flow through a clay membrane caused by an initial difference in the concentration of salt solutions in reservoirs on the two sides of the membrane. Transport is assumed to be controlled by three coefficients, which are analogous to the permeability, salt diffusivity, and salt reflection coefficient of the membrane. Initial fluid motion is caused by osmosis, leading to a buildup of pressure on one side of the membrane. However, the clay forms an imperfect ion exclusion membrane and the final steady state is one of equal concentrations and pressures on the two sides of the membrane. The time-dependent differences in pressure and salt concentration across the membrane are predicted to vary as the sum of two decaying exponentials. When the salt reflection coefficient is small, one time scale governs Darcy flow through the membrane and another the diffusion of salt. Experimental results confirm the analysis. Although the salt concentration in the reservoirs was monitored in the experiments, estimates of the transport coefficients can be obtained by measuring only the pressure change across the membrane. Copyright 2000 Academic Press. PMID- 11017744 TI - Electroosmosis through a Cation-Exchange Membrane: Effect of an ac Perturbation on the Electroosmotic Flow. AB - Electroosmosis experiments through a cation-exchange membrane have been performed using NaCl solutions in different experimental situations. The influence of an alternating (ac) sinusoidal perturbation, of known angular frequency and small amplitude, superimposed to the usual applied continuous (dc) signal on the electroosmotic flow has been studied. The experimental results show that the presence of the ac perturbation affects the electroosmotic flow value, depending on the frequency of the ac signal and on the solution stirring conditions. In the frequency range studied, two regions have been observed where the electroosmotic flow reaches a maximum value: one at low frequencies ( approximately Hz); and another at frequencies of the order of kHz. These regions could be related to membrane relaxation phenomena. Copyright 2000 Academic Press. PMID- 11017745 TI - Preparation and Characterization of Ceramic Hollow Microspheres for Heavy Metal Ion Removal in Wastewater. AB - Ceramic hollow microspheres (CHMSs) were prepared to use as supports for the removal of heavy metal ions from industrial waste-water. A water extraction sol gel technique was used to prepare porous CHMS by extracting water from an emulsion of LUDOX (silica colloid; SiO(2), Aldrich Co.) and 2-ethyl-1-hexanol. Experiments were conducted to control pore size, wall thickness, and separation yield by examining the ratio of precursors (LUDOX and 2-ethyl-1-hexanol), catalyst (NH(4)OH), sintering temperature, surfactant (SPAN 80), extractant (n butanol), stirring speed, and concentration of precursor (LUDOX). The results revealed that the optimum conditions were 20 ml of a 10 wt% solution of LUDOX, 10 ml of NH(4)OH, a sintering temperature of 500 degrees C, 0.4 ml of SPAN 80, 200 ml of n-butanol, and a stirring speed of 730 rpm/100 ml of 2-ethyl-1-hexanol. CHMSs were impregnated in Cyanex 272 and examined for their ability to remove heavy metal ions from a solution. Based on an experiment involving the removal of metal ions using CHMSs that were prepared under optimum conditions, Zn ion was removed at a level of 0.354 mmol/g at pH 4, which was about twice the adsorption capacity of CHMSs prepared by Wilcox (Mater. Res. Soc. Symp. Proc. 346, 201 (1994)). Copyright 2000 Academic Press. PMID- 11017746 TI - Theory of Frequency-Dependent Polarization of General Planar Electrodes with Zeta Potentials of Arbitrary Magnitude in Ionic Media. AB - Electrode polarization effects have long aggravated the efforts of low frequency analysis, particularly those investigations carried out on biological material or in highly conductive media. Beginning from elementary equations of electrostatics and hydrodynamics, a comprehensive model is devised to account for the screening of a general planar electrode by an ionic double layer. The surface geometry of the planar electrode is left unspecified to include any type of micromachined array. Building on the previous work by DeLacey and White (1982, J. Chem. Soc. Faraday Trans. 2 78, 457) using a variational theorem, we extend their numerical results with compact analytic solutions, analogous to the Debye-Huckel potential for dc systems, but applicable now to dynamic ac experiments. The variational approach generates functions that are not restricted by perturbation expansions or numerical convergence, representing optimal approximations to the exact solutions. Copyright 2000 Academic Press. PMID- 11017747 TI - Theory of Frequency-Dependent Polarization of General Planar Electrodes with Zeta Potentials of Arbitrary Magnitude in Ionic Media. AB - The expressions obtained in the previous paper for electrode polarization are applied to a homogeneous planar electrode and a planar array of electrodes used in the generation of nonuniform fields. The effective far field experienced outside the double layer is computed for both electrodes, and sample spectra are provided. The effective far field expression contains the electrode impedance and the effects of concentration polarization due to the static double layer on the electrode generated by the zeta potential. The effective far field results are compact and contain simple integrals that can be evaluated numerically. Copyright 2000 Academic Press. PMID- 11017748 TI - Highly Dispersed X/SiO(2) and C/X/SiO(2) (X=Alumina, Titania, Alumina/Titania) in the Gas and Liquid Media. AB - Fumed oxides, such as silica, alumina, titania, and mixed X/silicas (X=Al(2)O(3) (AS), TiO(2) (TS), CVD-TiO(2), Al(2)O(3)/TiO(2) (AST)), pristine or covered by carbon deposits formed due to pyrolysis of cyclohexene, were studied using nitrogen adsorption-desorption, photon correlation spectroscopy particle sizing, and electrophoresis. A significant influence of the nature of surface-active sites and structural features of oxides (individual silica, mixed fumed, or prepared using chemical vapor deposition (CVD)) on the pyrolysis of cyclohexene is observed with respect to the pore size distributions due to differences between primary particles in aggregates and on their outer surfaces in the filling of channels by pyrocarbon, resulting also in a decrease in fractal dimension. Structural characteristics and dependences of the particle size distribution and electrokinetic potential of X/SiO(2) and C/X/SiO(2) on the pH of aqueous suspensions suggest that the carbon deposit covers mainly acidic sites at the X/SiO(2) interfaces and X phase patches possessing catalytic activity in pyrolysis, as the negative charge of particles is reduced by pyrocarbon grafting. Copyright 2000 Academic Press. PMID- 11017749 TI - Electrooptics of beta-FeOOH Particles in Aqueous Media. AB - Reversing-pulse electric birefringence (RPEB) of a nearly monodisperse iron(III) hydroxide oxide sample in the beta-form (beta-FeOOH) was measured at 25 degrees C and at a wavelength of 633 nm in aqueous media in the presence of NaCl. The concentrations of beta-FeOOH and added NaCl varied between 0.00111 and 0.0555 g/L and 0.03 and 2.0 mM, respectively. Except for the suspensions with high salt concentrations, each RPEB signal showed a dip or minimum in the reverse process upon electric field reversal, together with a smooth rise in the buildup and a fall in the decay process. The observed signals were analyzed with a new RPEB theory, which takes into account not only the permanent electric dipole moment (u) but also the root-mean-square ionic dipole moment ((1/2)) due to the ion fluctuation in ion atmosphere, in addition to the field-induced electronic (covalent) dipole moment Deltaalpha' E. The results showed that the slowly fluctuating moment of (1/2) is by far the most predominant one for the field orientation of the beta-FeOOH particle, though the permanent dipole moment u may not be completely excluded. The rotational relaxation time of the whole particle was evaluated from the decay signal, while the relaxation time for fluctuating ions was estimated from RPEB signal fitting. The sign of the steady state birefringence for beta-FeOOH suspensions was positive without exception under the present conditions. The birefringence signals in the steady state (delta/d) were proportional to the second power of the applied field strength (E) in the low field region; thus, the Kerr law was verified to hold for beta-FeOOH suspensions. The specific Kerr constant was evaluated for each suspension by extrapolating the values of delta/d to zero field (E-->0). Copyright 2000 Academic Press. PMID- 11017750 TI - Wetting Study of Hydrophobic Membranes via Liquid Entry Pressure Measurements with Aqueous Alcohol Solutions. AB - A new concept of liquid entry pressure measurements is applied to study the hydrophobicity of microporous membranes for aqueous alcohol solutions. The effects of alcohol concentration, type of alcohol, and temperature on liquid entry pressure of the membrane have been studied. Two theoretical equations for the determination of membrane pore size have been proposed. The former equation was developed taking into account the deviation from the Laplace-Young equation due to the membrane structure by means of the structure angle. The latter equation was established considering only the range of alcohol concentration in which the dispersion component of liquid surface tension remains practically constant. Hydrophobicity has been expressed in terms of wetting surface tension, gamma(L)(w). Based on these measurements, the maximum concentration before the spontaneous wetting occurs would be predicted. Copyright 2000 Academic Press. PMID- 11017751 TI - H(2)O Outgassing Properties of Fumed and Precipitated Silica Particles by Temperature-Programmed Desorption. AB - Temperature-programmed desorption was performed at temperatures up to 850 K on as received fumed and precipitated silica particles. Physisorbed water molecules on both types of silica had activation energies in the range of 38-61 kJ/mol. However, the activation energies of desorption for chemisorbed water varied from approximately 80 to >247 kJ/mol for fumed silica, Cab-O-Sil-M-7D, and approximately 96 to 155 kJ/mol for precipitated silica, Hi-Sil-233. Our results suggest that physisorbed water can be effectively pumped away at room temperature (or preferably at 320 K) in a matter of hours. Chemisorbed water with high activation energies of desorption (>126 kJ/mol) will not escape silica surfaces in 100 years even at 320 K, while a significant amount of the chemisorbed water with medium activation energies (80-109 kJ/mol) will leave the silica surfaces in that time span. Most of the chemisorbed water with activation energies <126 kJ/mol can be pumped away in a matter of days in a good vacuum environment at 500 K. We had previously measured about 0.1-0.4 wt% of water in silica-reinforced polysiloxane formulations containing approximately 21% Cab-O-Sil-M-7D and approximately 4% Hi-Sil-233. Comparing present results with these formulations, we conclude that the adsorbed H(2)O and the Si-OH bonds on the silica surfaces are the major contributors to water outgassing from these types of silica-filled polymers. Copyright 2000 Academic Press. PMID- 11017752 TI - Application of the JKR Method to the Measurement of Adhesion to Langmuir-Blodgett Cellulose Surfaces. AB - The JKR method has been applied for studying adhesion between poly(dimethylsiloxane) (PDMS) caps and Langmuir-Blodgett cellulose surfaces including the substrate, hydrophobized mica, and two flat mineral surfaces, bare mica and glass. The self-adhesion of PDMS caps and oxidized PDMS caps are included as a reference to compare with literature data. The results of the measurements have been compared with previous studies using the surface force apparatus and similar systems. A satisfactory agreement is obtained for simple systems showing no, or very limited, hysteresis between loading and unloading curves. In several cases, however, a large hysteresis is found between loading and unloading curves, with a larger adhesion measured from the pull-off force than from the JKR-curve determined on loading. This is, for instance, the case for PDMS against cellulose. The situation is analogous to that found in wetting studies showing a large hysteresis between advancing and receding contact angles. Copyright 2000 Academic Press. PMID- 11017753 TI - Interactions between Poly(ethylene oxide) and Sodium Alkylcarboxylates As Studied by Conductivity and Gel Permeation Chromatography. AB - The interactions between PEO and sodium alkylcarboxylates (octyl, decyl, and dodecyl) have been investigated by conductivity measurements and gel permeation chromatography (GPC). Also included in the study was sodium dodecyl sulfate. From the conductivity measurements the critical aggregation concentration, ionization degree, and binding ratios were determined; the binding ratio was also determined from GPC. PEO-surfactant interactions were observed for all the studied surfactants, except sodium octanoate. For the polymer-surfactant complexes the ionization degree was in all cases observed to be about 0.2 higher than the ionization degree for the corresponding aqueous micelles. Further, the binding ratio decreased somewhat with decreasing chain length of the alkylcarboxylate. The Gibbs free energy showed that the polymer-surfactant interaction decreases with decreasing chain length of the alkylcarboxylates and is weaker for alkylcarboxylate compared to alkylsulfate of similar chain length. Copyright 2000 Academic Press. PMID- 11017754 TI - Menisci in a Diamond-Shaped Capillary. AB - This paper presents a closed form analytical solution to the augmented Young Laplace equation for the meniscus profile in a capillary formed between four equal-sized tangent cylinders centered on the vertices of a square. The solution is valid for a large class of disjoining pressure isotherms and contact angles. Copyright 2000 Academic Press. PMID- 11017755 TI - A comprehensive evaluation of the effects and mechanisms of antifreeze proteins during low-temperature preservation. AB - During the past 10 years, it has become clear that the effects of antifreeze proteins (AFPs) on cell viability and on thermodynamic properties during low temperature preservation are complex, even controversial. In this paper, these studies are reviewed systematically and some conclusions are drawn. It is shown that AFPs can display both protective and cytotoxic actions and both nucleation of ice and inhibition of ice crystal growth, depending on several factors; these include the specific storage protocol, the dose and type of AFP, the composition and concentration of cryoprotectant, and the features of the biological material. A novel model, incorporating some recent findings concerning these proteins, is proposed to explain this dual effect of AFPs during cryopreservation. AFP-ice complexes have some affinity interactions with cell membranes and with many other molecules present in cryopreservation solutions. When the intensity of these interactions reaches a certain level, the AFP-ice complexes may be induced to aggregate, thereby inducing ice nucleation and loss of the ability to inhibit recrystallization. PMID- 11017756 TI - Changes in the surface potential of Lactobacillus acidophilus under freeze thawing stress. AB - The zeta potential of Lactobacillus acidophilus CRL 640, a measure of the net distribution of electrical charges on the bacterial surface, is a function of the glucose concentration in the growing media. With 2% glucose, cells in the stationary phase showed a zeta potential of -45 +/- 2 mV. With these cells, the zeta potential after freezing and thawing decreased to -32 +/- 2 mV and there was a decrease in viability. The changes in the surface potential correlated with damage to the cell surface as shown by electron microscopy. Freeze-thawed cells incubated in a rich medium recovered a zeta potential of -38 +/- 2 mV without cell growth. L. acidophilus CRL 640 showed the same value of surface potential as control cells when they were frozen and thawed in 2 M glycerol. PMID- 11017757 TI - Stabilization and preservation of Lactobacillus acidophilus in saccharide matrices. AB - Lyophilization and vacuum- or spray-drying are some of the most useful techniques for preserving foods, agricultural products, and pharmaceuticals. Biological materials, however, can be irreversibly damaged during these treatments. Therefore, it is essential to design protective agents to preserve protein activity and cell viability. In this paper we examine the use of alpha, alpha trehalose-borate systems as protectants for Lactobacillus acidophilus during freeze- and vacuum-drying. Trehalose was found to be an effective protectant for freeze-dried and vacuum-dried samples, and it is equivalent to a protective formulation which is in current industrial use. It is known from our previous work on enzymes that the presence of borate can dramatically enhance the protective ability of trehalose. In this work, the addition of trehalose-borate to bacterial concentrate greatly improves the recovery of viable cells after storage. This improvement was seen in freeze-dried samples stored at 37 degrees C as well as for vacuum-dried samples held at room temperature. A tailored buffering strategy was tested to counteract the high pH resulting from the addition of borate to the mixture. Use of citric or lactic acids in combination with ammonium hydroxide gave a protectant solution with high pH (resulting in effective crosslinking between trehalose and borate) but a dry product with reduced pH upon rehydration (conducive to cell survival). These results raise exciting possibilities for protection of more labile prokaryotic species as well as simple eukaryotes. PMID- 11017758 TI - Overcoming a permeability barrier by microinjecting cryoprotectants into zebrafish embryos (Brachydanio rerio). AB - The goal of this research was to examine the developmental effects on zebrafish embryos (Brachydanio rerio) when cryoprotectants were directly microinjected into the yolk. Our objectives were to: (i) determine the final concentration of propylene glycol (PG) and dimethyl sulfoxide (Me(2)SO) that the embryos could tolerate without causing teratogenic effects; (ii) determine if the toxicity of Me(2)SO could be reduced by the simultaneous presence of various proportions of amides; and (iii) examine whether this intracellular cryoprotectant incorporation could reduce the cryodamage to the yolk syncytial layer (YSL) after vitrification trials. The rationale for conducting these microinjection experiments was to overcome the permeability barrier of the YSL. Intracellular PG produced better survival than Me(2)SO (P < 0.05). Embryos tolerated both 10- and 30-nl microinjections of PG, yielding final concentrations of 2.3 and 5.0 M within the yolk, resulting in 70 +/- 3 and 35 +/- 4% survival at day 5, respectively. In similar experiments with Me(2)SO, survival was lower than PG at 60 +/- 4 and 14 +/- 4% at 2.4 and 5.2 M. Unlike other cellular systems, the presence of amides, specifically acetamide or formamide, did not reduce the toxicity of Me(2)SO in zebrafish embryos (P > 0.05). During vitrification trials, we estimated a 25% dehydration of the yolk, yielding an effective PG concentration of 5.9 M. However, the incorporation of this vitrifiable concentration of PG was not sufficient to improve the postthaw morphology of the YSL (P > 0.05). Clearly, other factors need to be examined in establishing a successful vitrification protocol for zebrafish embryos. PMID- 11017759 TI - Effect of reduced concentrations of glycerol and various macromolecules on the cryopreservation of mouse and cattle embryos. AB - The effect of different macromolecules [bovine serum albumin (BSA), Pluronic F 68, (ET surfactant), or sodium hyaluronate (SH)] on postthaw survival of mouse morulae and in vivo- and in vitro-derived bovine blastocysts frozen in 10, 5, or 1% glycerol solutions was investigated. Embryos were equilibrated with cryoprotectant solution at 25 degrees C for 10 min, seeded at -5 degrees C, cooled at 0.5 degrees C/min to -35 degrees C, and plunged into liquid nitrogen. Embryos were thawed in a 35 degrees C water bath, glycerol was removed with 0.6 M sucrose at 25 degrees C for 5 min, and postthaw viability was evaluated after 1, 24, and 48 h in culture. The addition of BSA supplementation improved postthaw survival of mouse morulae frozen in 5% glycerol, but not in 10% glycerol. All three macromolecular supplements were effective in increasing survival of mouse morulae in 5% glycerol but only BSA and SH were effective in increasing postthaw survival of in vivo- and in vitro-derived bovine blastocysts. None of the macromolecular supplements improved postthaw survival of embryos frozen in 1% glycerol. PMID- 11017760 TI - Paradoxical effects of temperature on vascular tone. AB - Temperature may have significant influence on vascular tone in such cases as organ preservation, coronary bypass surgery, and extracorporeal circulation. The aim of this research was to study the direct effect of temperature variation on vascular tone in an attempt to elucidate the mechanisms involved. In a first series of experiments, the isometric tension of two different vessels (rat thoracic aorta and pig renal branch artery) was studied at different temperatures. To study the role of calcium in this response, a second series of experiments was performed. In this the vessels were incubated with the intracellular chelator BAPTA/AM. Further experiments were performed to test the effect of cold storage. Our results show that changes in temperature lead to different results in pig renal artery and rat aorta. A decrease in temperature induced a highly reproducible relaxation in rat aorta, whereas pig renal artery presented cooling-induced contraction. Moreover, whereas calcium depletion failed to inhibit cooling-induced relaxation in rat aorta, it did not provoke cooling induced contraction in pig renal artery. Similar responses were obtained with cold storage and calcium depletion. We intend to demonstrate that, just as the effect of temperature variation on pig renal artery is due to a metabolic mechanism, its effect on rat aorta may be due to structural factors. This hypothesis is supported by the result of histological studies which demonstrate a higher proportion of elastin fibres in rat aorta than in pig renal artery. PMID- 11017761 TI - Thyroid cryotherapy in an experimental rat model-topography of temperature during therapy and functional results. AB - The aim of this study was to investigate post cryotherapy thyroid function status of normal rat thyroid tissue and to determine the topography of temperature of cryotreated tissues and of tissues adjacent to them. Nitrous oxide cryotherapy was performed in 40 male Wistar rats. They were divided into four groups of 10. In group I, the right thyroid lobe was subjected to cryotherapy and the left lobe was not frozen. In group II, both thyroid lobes were cryotreated. In group III, the right lobe was frozen and the left lobe was excised. In group IV, the thyroid was subjected to neither cryotherapy nor surgery. During cryotherapy, the temperature in various places of the thyroid and in the surrounding tissues was measured. Serum thyrotropin concentrations were determined before an experiment and 4 weeks after in all rats. The results of temperature measurements proved that it is possible to limit cryotherapy to certain areas of thyroid tissue and to spare the neighboring tissues, because they are not subjected to temperatures that are damaging. The effectiveness of cryotherapy was confirmed by functional effect. Cryotherapy changed function of thyroid tissue. There was a statistically significant difference between mean baseline and follow-up concentrations in rats of groups II and III. In both groups hypothyroidism occurred post cryotherapy. PMID- 11017762 TI - Assembly of intercellular junctions in epithelial cell monolayers following exposure to cryoprotectants. AB - We investigated the effects of cryoprotectants (glycerol, propane-1, 2-diol, dimethyl sulfoxide) on the ability of epithelial cells to assemble intercellular junctions. Madin-Darby canine kidney cells (MDCK, type II) were grown in S-MEM containing only 5 micromol/L Ca(2+) to allow attachment of cells to the growth surface but not the development of the junctional complex. In a first set of experiments, cells were exposed to 10% v/v cryoprotectant at room temperature for 30 min. After removal of the cryoprotectant, [Ca(2+)] was increased to 1.8 mmol/L (Ca-switch) and the assembly of junctions was followed immunocytochemically and by monitoring transepithelial resistance (TER). In a second set of experiments, the development of junctions was followed in the presence of 1% cryoprotectant. Addition and removal of 10% cryoprotectant had little effect on the assembly of junctions following the Ca-switch, with TER peaking >300 ohm cm(2) after 24 h. Immunocytochemical staining showed recruitment to cell borders of components of tight junctions, adherens junctions, and desmosomes and the presence of a distinct circumferential bundle of actin filaments. In the presence of 1% cryoprotectant, there was a lag of more than 20 h before TER began to rise. There was then a progressive rise in TER in all three cryoprotectant groups, indicating junction assembly, albeit at a lower rate than that in the absence of cryoprotectant. These results suggest that exposure to cryoprotectants per se will not inhibit cellular repair mechanisms aimed at restoring the integrity of epithelial cell layers, but incomplete removal of cryoprotectant may delay repair. PMID- 11017763 TI - Ultrastructure and phagocytic activity of rat peritoneal macrophages exposed to low temperatures in vitro. AB - Hypothermia affects various components of the immune system, leading to impaired immune resistance. To examine the in vitro effect of low temperature on the ultrastructure and phagocytic function of rat peritoneal macrophages, cells were incubated at 4, 10, 24, and 37 degrees C for 60 min. Subsequently, their ultrastructure and capacity to engulf latex particles and generate superoxide anions were evaluated. The results showed a close inverse relationship between incubation temperature and ultrastructural changes, i.e., the lower the temperature, the higher the number of altered cells. In addition, at lower temperatures the number of cells capable of phagocytosis was reduced; the cells engulfed fewer particles per cell and generated less superoxide anions. These findings may be relevant for explaining the increased susceptibility to bacterial infections under hypothermic conditions. PMID- 11017764 TI - Bioenergetic targeting during organ preservation: (31)P magnetic resonance spectroscopy investigations into the use of fructose to sustain hepatic ATP turnover during cold hypoxia in porcine livers. AB - During liver preservation, ATP supplies become depleted, leading to loss of cellular homeostatic controls and a cascade of ensuing harmful changes. Anaerobic glycolysis is unable to prolong ATP production for a significant period because of metabolic blockade. Our aim was to promote glycolysis during liver cold hypoxia by supplying fructose as an additional substrate, compared to supplementation with an equivalent concentration of glucose. Porcine livers (two groups; n = 5 in each) were retrieved by clinical harvesting techniques and subjected to two cycles of cold hypoxia and oxygenated hypothermic reperfusion. In the second cycle of reperfusion, the perfusate was supplemented with either 10 mmol/L glucose (Group 1) or 10 mmol/L fructose (Group 2). During reperfusion in both groups, similar levels of ATP were detected by phosphorus magnetic resonance spectroscopy ((31)P MRS). However, during subsequent hypoxia, ATP was detected for much longer periods in the fructose-perfused group. The rate of ATP loss was sevenfold slower during hypoxia in the presence of fructose than in the presence of glucose (ATP consumption of -7.2 x 10(-3)% total (31)P for Group 1 versus -1.0 x 10(-3)% total (31)P for Group 2; P < 0. 001). The changes in ATP were mirrored by differences in other MRS-detectable intermediates; e.g., inorganic phosphate was significantly higher during subsequent hypoxia in Group 1 (45.7 +/- 2.7% total (31)P) than in Group 2 (33.7 +/- 1.1% total (31)P; P < 0. 01). High resolution MRS of liver tissue extracts demonstrated that fructose was metabolized mainly via fructose 1-phosphate. We conclude that fructose supplied by brief hypothermic perfusion may improve the bioenergetic status of cold hypoxic livers by sustaining anaerobic glycolysis via a point of entry into the pathway that is different from that for glucose. PMID- 11017765 TI - A comparison of Peronospora parasitica (Downy mildew) isolates from Arabidopsis thaliana and Brassica oleracea using amplified fragment length polymorphism and internal transcribed spacer 1 sequence analyses. AB - Amplified fragment length polymorphism (AFLP) fingerprints and internal transcribed spacer 1 (ITS1) sequences from 27 Peronospora parasitica isolates (collected from Arabidopsis thaliana or Brassica oleracea), 5 Albugo candida isolates (from the same hosts and from Capsella bursa-pastoris), and 1 Bremia lactucae isolate (from Lactuca sativa) were compared. The AFLP analysis divided the isolates into five groups that correlated with taxonomic species and, in most cases, with host origin. The only exception was a group consisting of A. candida isolates from both B. oleracea and C. bursa-pastoris. ITS1 sequence analysis divided the isolates into the same five groups, demonstrated the divergence between P. parasitica isolates from A. thaliana and B. oleracea, and, using previously published ITS1 sequences, clearly showed the relationship between A. candida isolates from different hosts. PMID- 11017766 TI - Protein expression and subcellular localization of the general purine transporter UapC from Aspergillus nidulans. AB - The uapC gene of Aspergillus nidulans belongs to a family of nucleobase-specific transporters conserved in prokaryotic and eucaryotic organisms. We report the use of immunological and green fluorescent protein based strategies to study protein expression and subcellular distribution of UapC. A chimeric protein containing a plant-adapted green fluorescent protein (sGFP) fused to the C-terminus of UapC was shown to be functional in vivo, as it complements a triple mutant (i.e., uapC(-) uapA(-) azgA(-)) unable to grow on uric acid as the sole nitrogen source. UapC-GFP is located in the plasma membrane and, secondarily, in internal structures observed as fluorescent dots. A strong correlation was found between cellular levels of UapC-GFP fluorescence and known patterns of uapC gene expression. This work represents the first in vivo study of protein expression and subcellular localization of a filamentous fungal nucleobase transporter. PMID- 11017767 TI - Characterization of the ABC transporter genes MgAtr1 and MgAtr2 from the wheat pathogen Mycosphaerella graminicola. AB - ATP-binding cassette (ABC) transporters are membrane-bound transporters involved in various physiological processes. In this paper we describe the cloning of the ABC transporter encoding genes MgAtr1 and MgAtr2 from the wheat pathogen Mycosphaerella graminicola (anamorph Septoria tritici). Both deduced proteins MgAtr1 and MgAtr2 are highly homologous to other fungal ABC transporters. RT-PCR revealed that the MgAtr2 mRNA population consists of partially and fully spliced transcripts. Putative substrates of ABC transporters, modulators of ABC transporter activity, and inducers of ABC transporter gene transcription were analyzed for their potential to induce expression of MgAtr1 and MgAtr2 in m. graminicola. The genes are differently upregulated by compounds such as the plant secondary metabolites eugenol and reserpine. Similar results are obtained for several antibiotics and the azole fungicides cyproconazole and imazalil. Moreover, a different expression pattern between yeast-like cells and mycelium of this dimorphic fungus was observed. These results indicate that MgAtr1 and MgAtr2 play a role in protection of m. graminicola against natural toxic compounds and xenobiotics. A putative role in protection against plant defense compounds during pathogenesis is suggested. PMID- 11017768 TI - Biosynthesis of glycoproteins in Candida albicans: activity of mannosyl and glucosyl transferases. AB - The enzymes dolichol phosphate glucose synthase and dolichol phosphate mannose synthase (DPMS), which catalyze essential steps in glycoprotein biosynthesis, were solubilized and partially characterized in Candida albicans. Sequential incubation of a mixed membrane fraction with increasing concentrations of Nonidet P-40 released a soluble fraction that transferred glucose from UDP-Glc to dolichol phosphate glucose and minor amounts of glucoproteins in the absence of exogenous dolichol phosphate. Studies with the soluble fraction revealed that some properties were different from those previously determined for the membrane bound enzyme. Accordingly, the soluble enzyme exhibited a twofold higher affinity for UDP-Glc and a sixfold higher affinity over the competitive inhibitor UMP, and the transfer reaction was fourfold more sensitive to inhibition by amphomycin. On the other hand, a previously described protocol for the solubilization of mannosyl transferases that rendered a fraction exhibiting both DPMS and protein mannosyl transferase (PMT) activities operating in a functionally coupled reaction was modified by increasing the concentration of Nonidet P-40. This resulted in a solubilized preparation enriched with DPMS and nearly free of PMT activity which remained membrane bound. DPMS solubilized in this manner exhibited an absolute dependence on exogenous Dol-P. Uncoupling of these enzyme activities was a fundamental prerequisite for future individual analysis of these transferases. PMID- 11017769 TI - Genetic evidence for somatic haploidization in developing fruit bodies of Armillaria tabescens. AB - Armillaria spp. have vegetative hyphae with diploid uninucleate cells, but the fruit bodies of many species contain clamped dikaryotic hyphae. Earlier observations suggest that somatic haploidization takes place in developing fruit bodies. To verify this, a uninucleate diploid cell was isolated from each of the 49 mating combinations between single-spore isolates of Armillaria tabescens and they were fruited. Twenty-four isolates produced fruit bodies with at least a partially dikaryotic subhymenium. Dikaryotic hyphae were isolated from fruit-body primordia and homokaryons were obtained by micromanipulation or by protoplasting. Approximately half of the isolates proved to represent recombinant mating types in respect to parent homokaryons, and most of them contained recombinant haploid DNA, based on random-amplified microsatellite markers. The results show that the nuclei in dikaryotic hyphae found in fruit bodies result from somatic haploidization. The mechanism of haploidization remains unclear. PMID- 11017770 TI - A single five-step desaturase is involved in the carotenoid biosynthesis pathway to beta-carotene and torulene in Neurospora crassa. AB - Phytoene desaturase Al-1 from Neurospora crassa was expressed in Escherichia coli and an active enzyme was isolated which catalyzed the stepwise introduction of up to five double bonds into the substrate phytoene. The major reaction products were 3, 4-didehydrolycopene and lycopene. Several of the desaturation intermediates, zeta-carotene, neurosporene, and lycopene, were also accepted as a substrate by Al-1. In contrast to the structurally related bacterial enzymes, the cofactor involved in the dehydrogenation reaction was NAD for Al-1. In situ competition with a neurosporene- and lycopene-converting hydratase and cyclase indicated that these enzymes can divert intermediates of the desaturation sequence. Based on the in vitro and in vivo results, the organization of the phytoene desaturase from N. crassa was proposed as an assembly of identical protein units which are responsible for the multistep reaction. However, the spatial arrangement should be loose enough to allow an exchange of individual intermediates in both directions in and out of this complex. Since gamma-carotene is not accepted as a substrate by Al-1, the formation of torulene must proceed exclusively by the cyclization of 3,4-didehydrolycopene. PMID- 11017771 TI - Filamentous growth of Saccharomyces cerevisiae is regulated by manganese. AB - The Candida albicans INT1 gene is a virulence factor that contributes to both adhesion and filamentous growth of the fungus. Expression of INT1 in the budding yeast Saccharomyces cerevisiae directs both adhesion and filamentous growth. Because Int1p contains two predicted divalent cation-binding motifs, we asked whether divalent cations are important for the role of Int1p in filament formation. In this study, we found that INT1-induced filamentous growth (I-IFG) is sensitive to the divalent cation chelator EDTA and that this EDTA sensitivity can be ameliorated by the addition of Mn(2+), but not Mg(2+) or Ca(2+) ions. The addition of MnCl(2) restored both the proportion of cells forming filaments and the length of filaments formed. Expression of INT1 in S. cerevisiae mutants that reduce the intracellular concentration of Mn(2+) did not affect I-IFG. Interestingly, the Mn(2+) dependence of I-IFG is not dependent upon the presence of the putative divalent cation-binding domains found in INT1. Rather, we found that polarized growth induced by mutations in CDC12 and CLA4 or by expression of excess SWE1 was also sensitive to EDTA treatment and was restored by the addition of MnCl(2) but not by the addition of CaCl(2). Thus, our results suggest that in S. cerevisiae polarized growth is dependent upon the presence of Mn(2+) ions. PMID- 11017772 TI - Evolution of the thyroid hormone-binding protein, transthyretin. AB - Transthyretin (TTR) belongs to a group of proteins, which includes thyroxine binding globulin and albumin, that bind to and transport thyroid hormones in the blood. TTR is also indirectly implicated in the carriage of vitamin A through the mediation of retinol-binding protein (RBP). It was first identified in 1942 in human serum and cerebrospinal fluid and was formerly called prealbumin for its ability to migrate faster than serum albumin on electrophoresis of whole plasma. It is a single polypeptide chain of 127 amino acids (14,000 Da) and is present in the plasma as a tetramer of noncovalently bound monomers. The major sites of synthesis of TTR in eutherian mammals, marsupials, and birds are the liver and choroid plexus but in reptiles it is synthesised only in the choroid plexus. The observation that TTR is strongly expressed in the choroid plexus but not in the liver of the stumpy-tailed lizard and the strong conservation of expression in the choroid plexus from reptiles to mammals have been taken as evidence to suggest that extrahepatic synthesis of TTR evolved first. The identification and cloning of TTR from the liver of an amphibian, Rana catesbeiana, and a teleost fish, Sparus aurata, and its absence from the choroid plexus of both species suggest an alternative model for its evolution. Protein modelling studies are presented that demonstrate differences in the electrostatic characteristics of the molecule in human, rat, chicken, and fish, which may explain why, in contrast to TTR from human and rat, TTR from fish and birds preferentially binds triiodo-l thyronine. PMID- 11017773 TI - Pattern of gonadal sex differentiation, development, and onset of steroidogenesis in the frog, Rana curtipes. AB - Histomorphological changes and steroidogenic potential of the gonads during sexual differentiation and development were studied in Rana curtipes from tadpole stage 25 (Gosner) until maturity. In stage 25 tadpoles of smaller snout-vent length (SVL; 4-5 mm) the gonads were indifferent, containing a few somatic and germ cells, whereas in larger tadpoles (SVL > 7 mm) gonads were differentiated into ovaries with a central lumen. Onset of meiosis was seen in these ovaries. At stage 26, diplotene and first growth phase oocytes were found. With advancement in developmental stage and after metamorphosis the ovaries progressively enlarged due to increase in number and size of oocytes. Vitellogenesis began in the ovary of 4-month-old frogs. Females attained maturity 6 months after metamorphosis. The frogs showed amplexus and one frog spawned. Onset of testicular formation seen at stage 31 was associated with the degeneration of oocytes and infiltration of darkly stained somatic cells in the central region. By stage 35 all oocytes degenerated, leaving behind a large number of somatic and germ cells interspersed with each other. At stage 38, formation of seminiferous tubules enclosing spermatogonia and pre-Sertoli cells was seen. Initiation of meiosis was evident at metamorphic climax. Cysts of elongated spermatids associated with Sertoli cells were seen in 45-day-old frogs. Histochemically, delta(5)-3 beta hydroxysteroid dehydrogenase activity was localized in the ooplasm, follicular cells, and interstitium of the ovary from stage 28 onward. The enzyme activity in the testis appeared in 45-day-old froglets. In R. curtipes gonadal differentiation is a semidifferentiated type since gonads initially differentiate into ovaries, and later, in the prospective males, the ovaries degenerate and transform into testes. The males attain maturity much earlier than the females. In R. curtipes gonadal sex differentiation precedes the onset of gonadal steroidogenesis. PMID- 11017774 TI - cDNA cloning and functional expression of growth hormone receptor from soft shelled turtle (Pelodiscus sinensis japonicus). AB - The growth hormone receptor (GHR) cDNA was cloned from the liver of soft-shelled turtle (Pelodiscus sinensis japonicus) using the polymerase chain reaction (PCR). Although GHR has been cloned from several mammalian and avian species, this is the first description of the reptilian receptor. As deduced from the nucleotide sequence, the precursor GHR of soft-shelled turtle (tGHR) is a protein of 615 amino acids which presents 72% identity with the chicken receptor and 57-64% identity with GHRs of several mammals. The tGHR expressed in COS-7 cells specifically bound human growth hormone (hGH) and was able to transduce an activation of transcription in the transfected cells. Binding of (125)I-hGH to the expressed receptor was decreased by the addition of excess unlabeled hGH, pig GH, and bream GH but not by pig insulin. The open reading frame of tGHR cDNA was inserted into the pSINrep/gfp (green fluorescence protein) vector and the tGHR gfp fusion protein was stably expressed in baby hamster kidney (BHK) cells. Confocal imaging showed that tGHR-gfp was largely concentrated on the plasma membrane. Western blot analysis and deglycosylation treatment with PNGase F demonstrated that tGHR was a glycoprotein in BHK cells. PMID- 11017775 TI - Spatio-temporal expression of TSH beta and FSH beta genes in normally metamorphosing, metamorphosed, and metamorphosis-arrested Hynobius retardatus. AB - Expression patterns of TSH beta and FSH beta genes were investigated in normally metamorphosing and metamorphosed individuals and in goitrogen-treated (TU-SPC), metamorphosis-arrested larvae of a Japanese salamander, Hynobius retardatus, a particular population of which had been reported to show neoteny. Cloned sequences for Hynobius TSH beta and FSH beta covered 66 and 56% of full-length genes, respectively, and both showed relatively high similarity to those of other vertebrates. TSH beta-expressing cells were observed at the ventro-caudal region of the pars distalis of controls, whereas an extraordinarily large number of cells which occupied the whole area of the pituitary glands strongly expressed TSH beta in TU-SPCs. In contrast, no FSH beta-expressing cells were observed in the pituitary glands of controls, whereas several positive cells were observed in the pituitary glands of TU-SPCs. Northern blot analyses revealed a rapid increase of TSH beta expression in the pituitary gland of TU-SPCs that finally reached the level of 20- to 30-fold as much as that of controls. Surprisingly, the expression of FSH beta also increased in the pituitary gland of TU-SPCs compared with that of controls. On the other hand, triiodothyronine administration to the control juveniles reduced the expression of TSH beta compared with saline-administrated controls, suggesting that thyroid hormone (TH) exerts a repressing effect on TSH beta gene. These results suggest that TSH beta overexpression in TU-SPCs must be caused by a deprivation of TH which plays a role in the negative feedback system and that FSH beta expression also might be enhanced by mechanisms similar to those of TSH beta, probably due to certain overlapping of the endocrinological regulation of TSH and FSH in H. retardatus. PMID- 11017777 TI - Fecal progesterone, estrogen, and androgen metabolites for noninvasive monitoring of reproductive function in the female Indian rhinoceros, Rhinoceros unicornis. AB - This investigation aimed to establish noninvasive methods for endocrine monitoring of estrous cycles and pregnancy in the Indian rhinoceros. Fecal samples were collected 1-3 times per week from nonpregnant and pregnant captive females (n = 7). Enzyme immunoassays for fecal progesterone, androgen, and estrogen metabolites, respectively, were tested for their ability to determine follicular and luteal phases and to characterize endocrine profiles during pregnancy. Antibodies used were raised against pregnanediol (20 alpha-OH pregnanes), 20-oxo-pregnanes, epiandrosterone (17-oxo-androstanes), and total estrogens. Androgens and estrogens were found to be reliable indicators of the follicular phase, whereas 20 alpha-OH- and 20-oxo-pregnanes were reliable indicators of luteal function. Progesterone metabolites were also reliable indicators of pregnancy, whereas 17-oxo-androstanes and estrogens were basal throughout gestation. Estrous cycles were regular throughout the year, with an average cycle length of 43.4 +/- 1.5 (n = 27) days; the length of the follicular phase, as indicated by elevated estrogen levels, was 15.9 +/- 1.0 days, whereas the luteal phase, as indicated by elevated 20-oxo-pregnane levels, was 19.1 +/- 0.4 days. Fecal pregnane values were already increasing while follicular estrogen values were still decreasing. The length of the diestrus, indicated by basal steroid levels between declining 20-oxo-pregnanes and subsequently increasing estrogens, was 11.4 +/- 1.2 days. Pregnane levels increased from the 3rd month of gestation onward and levels exceeded luteal phase concentrations approximately 10 times by the 7th month of gestation onward. HPLC separation of immunoreactive fecal metabolites indicated the presence of estrone, estradiol-17beta, and several 17-oxo-androstanes, 20 alpha-OH-pregnanes, and 20-oxo-pregnanes. Concentrations of a peak with an elution profile similar to that of pregnanediol increased as pregnancy progressed. Postpartum fecal estrogen and 17-oxo androstane concentrations in one animal indicated follicular development comparable to the follicular phase of the estrous cycle, but this was not followed by a subsequent luteal phase. In conclusion, estrous cycle and pregnancy in Indian rhinoceroses can be monitored using fecal steroid analysis. Pregnane metabolites were reliable indicators of the corpus luteum and pregnancy, whereas fecal 17-oxo-androstanes and estrogens were indicators of the follicular phase. PMID- 11017776 TI - Binding of xenobiotics to hepatic estrogen receptor and plasma sex steroid binding protein in the teleost fish, the common carp (Cyprinus carpio). AB - Competitive receptor binding assays have been suggested as an in vitro screening tool for assessing the activity of alleged estrogenic substances. In this study, we determined the ability of steroidal and nonsteroidal substances to inhibit the binding of [(3)H]17 beta-estradiol (E2) to the hepatic estrogen receptor (ER) and the plasma sex steroid binding protein (SBP) of the teleost fish, the common carp (Cyprinus carpio). The objectives of the study were (1) to characterize ER binding in the liver cytosol of male and female carp, (2) to establish complete [(3)H]E2 displacement curves from carp ER for a range of natural and xenobiotic substances and to compare the ligand data of carp ER with published data from other vertebrate species to reveal possible species differences, and (3) to determine the interaction of natural and xenobiotic substances with the steroid binding site of SBP in carp plasma. The results indicate the presence of a single class of estrogen binding sites with high affinity and limited capacity in liver cytosol of carp. The various test agents showed partly quantitative differences in their binding affinities, with the xenobiotics generally showing limited ability to displace [(3)H]E2 from the hepatic ER or from plasma SBP of carp. However, we found no evidence that a compound is an ER ligand exclusively in one species. The findings of this study indicate that interspecies extrapolation of steroid receptor binding data is possible on a yes/no basis but not on a quantitative basis. PMID- 11017778 TI - Estradiol regulation of follistatin and inhibin alpha- and beta(B)-subunit mRNA in avian granulosa cells. AB - Estradiol modulation of granulosa cell growth and regulation of follistatin and inhibin alpha- and beta(B)-subunit mRNA were investigated in cultured chicken granulosa cells. Granulosa cells were isolated and pooled according to size from the F(4) + F(5), small yellow (SYF), and large white (LWF) follicles. Isolated and dispersed granulosa cells were then cultured in the absence or presence of 1 x 10(-5) M 17 beta-estradiol. In Experiment 1 (n = 4 replications) the effect of estradiol on the growth of granulosa cells from the different-sized follicles was examined at 24 and 48 h of culture. Untreated and treated granulosa cells from all three follicle sizes proliferated during culture, and cell viability for all cultures was over 95% throughout the experiment. After 48 h the untreated cultures for all follicle types had 1.6 to 2.2 times (P < 0.05) more cells than the estradiol-treated cultures. In Experiment 2 (n = 3 replications), the cultures were terminated at 4 and 24 h after plating. Follistatin mRNA levels were higher in estradiol-treated cells at 24 h in F(4) + F(5) follicles, at 4 and 24 h in the SYF, and at 4 h in the LWF. beta(B)-subunit mRNA levels were also increased by estradiol at 4 h in F(4) + F(5) cells and at 4 and 24 h in the LWF. Steady state mRNA levels for the alpha-subunit were higher (P < 0.05) in estradiol-treated cultures at 4 and 24 h in F(4) + F(5) follicles and at 24 h in the SYF. Immunoreactive alpha-subunit protein, however, was not increased by estradiol treatment. Thus, whereas estradiol inhibited granulosa cell growth, it exerted a generally stimulatory effect on the expression of FS and the inhibin alpha- and beta(B)-subunit mRNA. PMID- 11017779 TI - Evidence for gonadotropin-releasing hormone-like peptides in a cnidarian nervous system. AB - There is increasing evidence that peptides of the gonadotropin-releasing hormone (GnRH) family, long considered a vertebrate preserve, are also present in invertebrate (molluscan) nervous systems. The possibility was examined that GnRHs are present and bioactive in cnidarians, considered to be representatives of the most primitive animals possessing a nervous system. Immunoreactive GnRH was detected in endodermal neurons of two anthozoans, the sea pansy Renilla koellikeri and the sea anemone Nematostella vectensis. In the sea pansy, immunoreactivity was detected throughout the autozooid polyps, including gamete producing endoderm. High-performance liquid chromatography and radioimmunoassays of extracts from whole sea pansy colonies yielded two elution peaks exhibiting GnRH immunoreactivity with antisera raised against shark or mammalian GnRH. Vertebrate GnRHs as well as the two sea pansy GnRH-like factors inhibited the amplitude and frequency of peristaltic contractions in the sea pansy, and these actions were blocked by the LHRH analog [D-pGlu(1),D-Phe(2),D-Trp(3,6)]-LHRH. These results suggest that the GnRH family of neuropeptides is more widespread in metazoans than previously thought. Although our physiological data are preliminary, they point to a role for GnRHs as inhibitory modulators of neuromuscular transmission in the sea pansy. PMID- 11017780 TI - Effect of diethylstilbestrol on thyroid hormone binding to amphibian transthyretins. AB - Transthyretin (TTR) is responsible for a major part of the binding of thyroid hormone to proteins in the plasma in amphibian tadpoles. To characterize the binding properties of amphibian TTRs, the effects of 17 hydrophobic signaling molecules, including 6 endocrine disruptors, on 3,5,3'-l-[(125)I]triiodothyronine ([(125)I]T(3)) binding to plasma proteins were examined in bullfrog Rana catesbeiana tadpoles. T(3) was the most potent competitive inhibitor among the 11 natural biological ligands studied, with an ID(50) of 8 nM. Diethylstilbestrol (DES) was the most powerful inhibitor among the 6 endocrine disruptors studied, with an ID(50) of 20 nM. Similar inhibitions of [(125)I]T(3) binding by these compounds were obtained when purified recombinant Xenopus and Rana TTRs were analyzed. Scatchard analysis revealed that Xenopus and Rana TTRs each possessed a single class of binding site for T(3), with a K(d) of 262 and 1.9 nM, respectively, at 0 degrees C. DES, at a concentration of 200 nM, induced the uptake of [(125)I]T(3) into Rana red blood cells suspended in Rana plasma from prometamorphic stages XIII-XV, when TTR is present in plasma. DES induced the uptake of [(125)I]T(3) into red blood cells to a lesser extent when they were suspended in Rana plasma from metamorphic climax stage XXIV, in which the level of TTR was lower than in plasma from the prometamorphic tadpoles. These results indicate that amphibian TTRs have the ability to bind DES with similar affinity to T(3), the natural ligand, and raise the possibility that DES binding to TTR might induce the temporary elevation of the free concentration of plasma T(3) followed by acceleration of cellular T(3) uptake. PMID- 11017781 TI - Involvement of crustacean hyperglycemic hormone in the control of gill ion transport in the crab Pachygrapsus marmoratus. AB - Total extracts of sinus glands (SG) of the euryhaline grapsid crab Pachygrapsus marmoratus contain peptidic factor(s) that stimulate osmoregulatory processes in isolated and perfused posterior gills from crabs acclimated to dilute seawater. This study investigated the nature of the active factor(s). Separation of P. marmoratus SG peptides by reverse-phase HPLC, followed by a direct enzyme-linked immunosorbent assay using an anti-Carcinus maenas crustacean hyperglycemic hormone (CHH) antiserum, identified a major immunoreactive chromatographic peak. A glucose quantification bioassay demonstrated a strong and specific hyperglycemic activity following injection of the immunoreactive peak, therefore defined as the CHH of P. marmoratus. Isolated posterior gills were then perfused with HPLC fractions using a dose of 4 SG equivalents/assay. The CHH fraction consistently and significantly increased the transepithelial potential difference and Na(+) influx by about 50%. The effect was rapid and reversible. Another substance of unknown nature (eluted earlier than CHH in the HPLC gradient) caused a small increase in Na(+) influx (14%) but had no effect on the transepithelial potential difference. No other peptidic product from the SG had significant effect on the measured osmoregulatory parameters. These results indicate that CHH, in addition to its hyperglycemic activity, is also implicated in the control of branchial ionic transport. This neuropeptide may thus constitute a major factor involved in the control of osmoregulation in decapod crustaceans. PMID- 11017782 TI - Three classes of cell surface receptors for alphaherpesvirus entry. PMID- 11017783 TI - The CD4-mediated immune response is critical in determining the outcome of infection using Theiler's viruses with VP1 capsid protein point mutations. AB - Daniel's strain of Theiler's virus (DA) induces a chronic demyelinating disease in the central nervous system (CNS) of susceptible SJL mice, which serves as an excellent model of multiple sclerosis. We previously demonstrated that point mutations near a putative virus receptor-binding site [VP1 99 (Gly to Ser) or 100 (Gly to Asp)] totally attenuate the ability of DA to persist and induce demyelination in SJL mice. The current studies demonstrate that class II restricted CD4(+) T cells play a major role in clearing VP1 mutant DA viruses from the CNS to prevent demyelination. Infection of SJL CD4((-/-)) mice with DA VP1-99(Ser) or DA-VP1-100(Asp) resulted in virus persistence and prominent demyelination in the spinal cord. In contrast, infection of SJL CD8((-/-)) mice with DA-VP1-99(Ser) or DA-VP1-100 did not result in virus persistence or demyelination. In addition, no virus-specific cytotoxicity was observed in CNS infiltrating lymphocytes following infection of SJL mice with VP1 mutant viruses. The mutant DA-VP1-99(Ser) and DA-VP1(100) viruses were in fact neurovirulent when compared to the wild-type DA virus, as they induced an overwhelming encephalitis and early lethality (2 to 4 days postinfection) in mice deficient in the IFN alpha/beta receptor. Therefore, the nondemyelinating phenotype observed with DA VP1-99(Ser) and DA-VP1-100(Asp) viruses is dependent in part on the CD4-mediated host immune response. PMID- 11017784 TI - Rubella virus capsid protein induces apoptosis in transfected RK13 cells. AB - Rubella virus is an enveloped positive-strand RNA virus that can cause mild to severe birth defects or death in an infected fetus. RV induction of programmed cell death, demonstrated in cell culture, has been implicated in the pathogenesis. The timing of apoptosis, 48 h p.i., suggested that accumulation of RV structural proteins might induce cell death in infected cells. Expression of RV structural proteins, capsid, envelope glycoproteins E1 and E2, in transiently transfected RK13 cells was as potent an inducer of cell death as RV infection. Immunofluorescence microscopy revealed that RV structural protein transfected cells exhibited the condensed nuclei typical of apoptotic cell death. Transfection with the capsid protein construct, but not E2 and E1, resulted in as much cell death as joint expression of all three RV structural proteins. Capsid required a membrane-anchoring domain to induce cell death, but a heterologous polypeptide fused to the capsid membrane anchor did not cause apoptosis. Deletion mutants demonstrated that the apoptosis-inducing activity resides in the N terminal 170 amino acids of capsid. Though apoptosis-inducing capsid constructs appear to have an ER sub-cellular localization, disruption of the ER calcium storage capacity does not correlate with cell death. Mechanisms consistent with these results are discussed. PMID- 11017785 TI - A novel baculovirus envelope fusion protein with a proprotein convertase cleavage site. AB - The entry mechanism of Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV), a group II NPV, in cultured cells was examined. SeMNPV budded virus (BV) enters by endocytosis as do the BVs of the group I NPVs, Autographa californica (Ac) MNPV and Orgyia pseudotsugata (Op) MNPV. In group I NPVs, upon infection acidification of the endosome triggers fusion of the viral and endosomal membrane, which is mediated by the BV envelope glycoprotein GP64. However, the SeMNPV genome lacks a homolog of GP64 envelope fusion protein (EFP). A functional homolog of the OpMNPV GP64 EFP was identified in SeMNPV ORF8 (Se8; 76 kDa) and appeared to be the major BV envelope protein. Surprisingly, a 60-kDa cleavage product of this protein is present in the BV envelope. A furin-like proprotein convertase cleavage site (R-X-K/R-R) was identified immediately upstream of the N-terminus of the mature Se8 protein and this site was also conserved in the Lymantria dispar (Ld) MNPV homolog (Ld130) of Se8. Syncytium formation assays showed that Se8 and Ld130 alone were sufficient to mediate membrane fusion upon acidification of the medium. Furthermore, C-terminal GFP fusion proteins of Se8 and Ld130 were primarily localized in the plasma membrane of insect cells. This is consistent with their fusogenic activity and supports the conclusion that the Se8 gene product is a functional homolog of the GP64 EFP. PMID- 11017786 TI - The fitness of citrus tristeza virus defective RNAs is affected by the lengths of their 5'- and 3'-termini and by the coding capacity. AB - Populations of the Closterovirus Citrus tristeza virus (CTV) generally contain defective RNAs (dRNAs) that vary in size, abundance, and sequence. The variation in abundance of the different dRNAs in a population suggests selection for those of higher fitness. To examine factors affecting fitness of dRNAs, we investigated a series of in vitro constructed dRNAs for their ability to be amplified in protoplasts by an efficiently replicated CTV deletion mutant. The minimal sequences required for accumulation of the dRNAs were within the genomic 5' proximal approximately 1 kb and the 3' 270 nucleotides. However, other factors were involved, because a dRNA with only the minimal sequences failed to be replicated. Rescue of a nonviable dRNA by insertion of nonviral sequences between the termini suggested that "spacing" between terminal cis-acting signals influenced fitness. A continuous open reading frame (ORF) through most of the sequences derived from the 5' of the genome was a requirement for dRNA amplification. In general, insertions, deletions, or nucleotide substitutions were tolerated in the dRNAs as long as an ORF was retained, whereas dRNAs with mutations that prematurely terminated the ORF were not viable. To discriminate between a requirement for an essential protein and ribosomal travel, perhaps to present replication signals to the replicase complex, mutations were made to modify the potential protein but still maintain an ORF. Deletions, insertions of nonviral sequences, or switching of reading frames that altered the amino acid sequence of the protein, except the N-terminal 161 amino acids, did not destroy the fitness of the dRNAs. Yet termination of the ORF in the middle of nonviral sequences did destroy the ability of the dRNAs to be amplified. These results suggest that even though a continuous ORF was needed for fitness, its protein product did not affect the amplification of the dRNAs. PMID- 11017787 TI - A protective role of interleukin-15 in a mouse model for systemic infection with herpes simplex virus. AB - To define the role of cytokine binding to the IL-2/IL-15R beta chain in protective immunity against systemic infection with herpes simplex virus type 2 (HSV-2), IL-2/IL-15 receptor(R)beta knock-out mice were inoculated intraperitoneally with HSV-2 strain 186. IL-2/IL-15R beta-deficient mice were susceptible to systemic HSV-2 infection compared with their heterozygous littermates. The emergence of natural killer (NK) cells was impaired in IL-2/IL 15R beta knock-out mice, but CD4(+) T cell receptor (TCR) alpha beta(+) T cells were normally detected in the peritoneal cavity after infection with HSV-2. However, the generation of HSV-2-specific CD4(+) T helper (Th) 1 cells producing interferon-gamma was impaired in IL-2/IL-15R beta knock-out mice following HSV-2 infection. The serum IL-15 level in control mice was increased in the early stage after HSV-2 infection but was not detectable in IL-2/IL-15R beta knock-out mice. In vivo administration of recombinant IL-15 protected normal mice from HSV-2 induced lethality, accompanied by increases in numbers of NK cells and HSV-2 specific Th1 cells. Taken together, these results suggest that IL-15, using the IL-2/IL15R beta chain, plays an important role in mounting protective immunity during the course of systemic HSV-2 infection. PMID- 11017788 TI - Characterization of epitopes for virus-neutralizing monoclonal antibodies to Ross River virus E2 using phage-displayed random peptide libraries. AB - Ross River virus (RRV) is the predominant cause of epidemic polyarthritis in Australia, yet the antigenic determinants are not well defined. We aimed to characterize epitope(s) on RRV-E2 for a panel of monoclonal antibodies (MAbs) that recognize overlapping conformational epitopes on the E2 envelope protein of RRV and that neutralize virus infection of cells in vitro. Phage-displayed random peptide libraries were probed with the MAbs T1E7, NB3C4, and T10C9 using solution phase and solid-phase biopanning methods. The peptides VSIFPPA and KTAISPT were selected 15 and 6 times, respectively, by all three of the MAbs using solution phase biopanning. The peptide LRLPPAP was selected 8 times by NB3C4 using solid phase biopanning; this peptide shares a trio of amino acids with the peptide VSIFPPA. Phage that expressed the peptides VSIFPPA and LRLPPAP were reactive with T1E7 and/or NB3C4, and phage that expressed the peptides VSIFPPA, LRLPPAP, and KTAISPT partially inhibited the reactivity of T1E7 with RRV. The selected peptides resemble regions of RRV-E2 adjacent to sites mutated in neutralization escape variants of RRV derived by culture in the presence of these MAbs (E2 210 219 and 238-245) and an additional region of E2 172-182. Together these sites represent a conformational epitope of E2 that is informative of cellular contact sites on RRV. PMID- 11017789 TI - Attachment of coxsackievirus B3 variants to various cell lines: mapping of phenotypic differences to capsid protein VP1. AB - The coxsackievirus B3 (CVB3) strain Nancy P establishes a persistent carrier state infection without visible cytopathic effect in primary human fibroblasts (HuFi H), whereas the derivative variant PD induces a complete lysis of the cell monolayer. To define the molecular basis of this exceptional growth property, the complete genomes of both viruses were sequenced and compared to all published sequences of CVB3. As a result, six unique amino acid substitutions in the VP1 capsid protein were observed. Via hybrid virus construction, the lytic phenotype was transferred to a nonlytic cDNA-generated CVB3. Mapping experiments indicate that the presence of amino acid residues K78, A80, A91, and I92 in VP1 is sufficient to induce "lytic" infections in HuFi H cells. Binding assays demonstrate that CVB3 Nancy P preferentially binds to the human coxsackievirus adenovirus receptor (CAR), while PD exhibits a very weak interaction with CAR but strong binding to the decay accelerating factor (DAF). These results suggest that the mutated amino acid residues in VP1 are involved in receptor recognition/binding. Moreover, the lytic replication of CVB3 PD and the hybrid virus in various nonpermissive rodent cell lines indicates that cell surface molecules other than CAR and DAF may be involved in attachment of this variant to cell surfaces. PMID- 11017790 TI - Rotavirus infection of MA104 cells is inhibited by Ricinus lectin and separately expressed single binding domains. AB - Various lectins were tested for blocking rotavirus infection of MA104 cells and it was observed that galactose-specific lectins were the most inhibitory. Of these Ricinus agglutinin was able to inhibit infection (by human and animal strains) at concentrations as low as 10(-9) M. In addition, in a virus overlay protein blot assay Ricinus agglutinin competed with simian rotavirus SA11 for binding to solubilized MA104 proteins. Amino acid sequence comparisons revealed similarity between the ricin toxin B subunit (which contains two separate carbohydrate-binding motifs: single binding domains (SBD) 1 and 2) and rotavirus spike protein VP4. A filamentous phage display system was used to independently express the two binding domains and while SBD1 inhibited infection of MA104 cells by CRW8, NCDV, and to a lesser extent Wa, SBD2 blocked only CRW8 and NCDV infection. Furthermore inhibition of CRW8 infection was a direct result of phage inhibiting virus attachment to cells. When amino acid 248 within SBD2 was mutated from the ricin toxin to the Ricinus agglutinin sequence this phage clone showed reduced binding to galactose and was no longer able to inhibit virus infection. Thus, rotavirus recognizes galactose as an important component of the receptor on MA104 cells. PMID- 11017791 TI - Mouse mammary tumor virus superantigen expression is reduced by glucocorticoid treatment. AB - Expression of mouse mammary tumor virus (MMTV)-encoded superantigens in B lymphocytes are required for viral transmission and pathogenesis. Due to problems with detection and quantification of the superantigen protein, most reports about the mechanism of superantigen expression from the viral sag gene rely on the quantitative analysis of putative sag mRNAs. The description of multiple promoters as a source of putative sag mRNA has complicated the situation even further. All conclusions about the level of superantigen protein expression based on these data remain circumstantial. To test the effect of the glucocorticoid hormone dexamethasone on the total superantigen expression from an infectious MMTV provirus we used a quantitative assay that is based on a superantigen luciferase fusion protein. MMTV gene expression from the major promoter in the 5' long terminal repeat (LTR) is strongly induced in the presence of glucocorticoid hormones. We now demonstrate that, in the presence of dexamethasone, sag gene expression is reduced despite increased transcription from the MMTV 5' LTR and increased amounts of putative sag mRNA initiated at the LTR promoter. These data show that the expression of the MMTV sag gene does not correlate with the activity of the major LTR promoter and thus differs from all other MMTV genes. PMID- 11017792 TI - Drug-resistant reverse transcriptase genotyping and phenotyping of B and non-B subtypes (F and A) of human immunodeficiency virus type I found in Brazilian patients failing HAART. AB - Development of drug resistance is the inevitable consequence of incomplete suppression of virus plasma levels in HIV-1-infected patients treated with highly active antiretroviral therapy. Resistance mutations previously characterized have been found in B subtype viruses of developed countries. Moreover, mutation profiles for non-B and more divergent B subtype viruses found in developing countries shall be analyzed together with their ex vivo phenotyping in order to establish an exact correlation between the genotyping data and the clinical management counseling for those uncommon virus subtypes. In the present study, we evaluated the mutation profile for individuals infected with B subtype and non-B subtype viruses. Viral DNA fragments corresponding to the RT gene were amplified, sequenced, and subtyped. Phenotyping analysis for reverse transcriptase nucleoside (NRTI) and nonnucleoside inhibitor susceptibility was performed using the recombinant virus assay technology. Brazilian non-B subtypes (subtype F, n = 4, and subtype A, n = 1) isolates showed essentially the same B subtype mutation profile, presenting an NRTI drug resistance with similar MIC50% and MIC90% values for all drugs analyzed regardless of their subtypes. A strong cross-resistance phenotype among AZT, 3TC, and abacavir could be seen in all isolates analyzed. A novel result was that some RT sequences not only revealed the presence of G333D/E mutations but also correlated to the presence of mutation T386I that could abrogate the M184V-surpassing effect of L210W or L210W plus G333D/E. These findings suggest that Brazilian non-B subtype HIV-1 strains use an identical RT drug resistance mutation pattern when compared to B isolates and will contribute to the validation of the genotypic and phenotypic tests in these predominant worldwide-spread viral variants. PMID- 11017793 TI - DNA vaccination of macaques by a full genome HIV-1 plasmid which produces noninfectious virus particles. AB - In this study, we tried a DNA vaccination regime in rhesus macaques using a full genome HIV-1 plasmid. The HIV-1 genome is under the control of its original LTR promoter, but has a mutated zinc finger motif gene in the nucleocapsid region. Due to the lack of genomic RNA packaging, the plasmid produces only noninfectious viral particles. We repeatedly injected four macaque monkeys intramuscularly with the naked DNA over a period of 40 weeks. To evaluate the humoral and cell mediated immunity provided by this DNA vaccination, no other booster or other recombinant viral vectors were used. Immunological responses against HIV-1 were elicited in all of the vaccinated monkeys: stable anti-HIV-1 Env antibodies were raised in two monkeys and CTL activities were induced in the other monkeys. The macaques were intravenously challenged at 54 weeks with 100 TCID(50) of SHIV-NM 3rN, which possesses an envelope gene homologous to the one in the vaccinated plasmid. In all of the vaccinated macaques, the peak plasma viral loads induced by the challenge virus were two to three orders of magnitude lower than those of the naive controls. These results suggest that a DNA vaccination regime with a full genome plasmid alone is potentially efficacious and provides a new possibility for the development of an AIDS vaccine. PMID- 11017794 TI - Mutations selected in rotavirus enterotoxin NSP4 depend on the context of its expression. AB - The rotavirus NSP4 protein is cytotoxic when transiently expressed in cells and is capable of inducing secretory diarrhea in neonatal mice. NSP4 consists of 175 amino acids, and sequences important for its toxic effects have been mapped to the carboxy-terminal half of the protein. In this report, we compared NSP4 encoding nucleotide sequences recovered from cell lines engineered to express NSP4 from human rotavirus strain Wa with NSP4 sequences recovered from cells persistently infected with either Wa or simian rotavirus strain SA11. In cells stably transfected with Wa NSP4, we found that proline(138) was changed to either serine or threonine. However, in cells persistently infected with SA11, we found that phenylalanine(33) was changed to leucine, and in cells persistently infected with Wa, no changes were observed in NSP4. Expression of Wa NSP4 in Caco-2 cells resulted in increased cell-doubling times and decreased cell viability in comparison to cells expressing NSP4-serine(138) or NSP4-threonine(138). This result suggests that sequence polymorphism at residue 138 in Wa NSP4 influences the cytotoxicity of the protein. Therefore, mutations in the carboxy-terminal half of NSP4 are selected when NSP4 is expressed in cells in the absence of other viral proteins, but not in the context of viral replication. These findings suggest that cytotoxic functions of NSP4 are not operant during natural rotavirus infection. PMID- 11017795 TI - In vitro assembly of bacteriophage P4 procapsids from purified capsid and scaffolding proteins. AB - Bacteriophage P4 is a satellite virus of bacteriophage P2, which has acquired the ability to utilize the structural gene products of P2 to assemble its own capsid. The normal P2 capsid has a T = 7 icosahedral structure comprised of the gpN derived capsid protein, whereas the capsid produced under the control of P4 has a smaller, T = 4 structure. The protein responsible for this size determination is the P4-coded gene product Sid, which forms an external scaffold on the P4 procapsid. Using an in vitro assembly system, we show that gpN and Sid can coassemble into procapsid-like particles, indistinguishable from those produced in vivo, in the absence of any other gene products. The fidelity of the assembly reaction is enhanced by the inclusion of PEG and has a pH optimum between 8.0 and 8.5. Analysis of the assembly properties of truncated versions of Sid and gpN suggests that the amino-terminal part of Sid is involved in gpN binding, while the carboxyl-terminal part forms trimeric Sid-Sid interactions, and that the first 31 amino acids of gpN are required for binding to Sid as well as for size determination. PMID- 11017796 TI - Human p32: a coactivator for Epstein-Barr virus nuclear antigen-1-mediated transcriptional activation and possible role in viral latent cycle DNA replication. AB - The Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is required for the maintenance of the viral chromosome in latently infected, proliferating cells and plays a role in latent cycle DNA replication. EBNA-1 also functions as a positive and negative regulator of EBV gene expression. We have investigated the interaction of EBNA-1 with p32, a host mitochondrial protein that associates with EBNA-1 in EBV-positive Burkitt's lymphoma cells. Using a chromatin immunoprecipitation assay, we found that a fraction of p32 localizes to the viral latent cycle origin of DNA replication oriP in vivo. p32 binds EBNA-1 independently of other proteins or DNA. EBNA-1 variants lacking one of two p32 binding elements did not interact stably with p32 in cultured cells and were defective for both transcriptional activation of a reporter gene linked to oriP FR and replication and/or maintenance of a plasmid bearing oriP. These results support a role for p32 in transcriptional activation by EBNA-1 and suggest that p32 plays a role in EBV latent cycle DNA replication. PMID- 11017797 TI - Characterization of the humoral immune response and virus replication in cats experimentally infected with feline foamy virus. AB - Cats were experimentally infected with cell culture-adapted feline foamy virus (FFV, spumaretrovirinae) isolate FUV. FFV was consistently recovered from peripheral blood leukocytes and throat samples of FFV-infected cats starting 2 to 3 weeks postinfection (p. i.), indicative of the establishment of persistent FFV infections. Viral persistence was established, even despite neutralizing antibodies that appeared early after infection. The humoral immune response toward FFV was quantitatively and qualitatively analyzed over time. FFV Gag specific antibodies were first detected 2 weeks p. i. and increased further; reactivities to the other structural and nonstructural FFV proteins appeared slightly delayed. Reactivities against FFV Pol and Gag proteins were detectable by immunoblotting and radioimmunoprecipitation, whereas the latter techniques had to be employed for the unambiguous detection of FFV Env-, Bet-, and Bel 1 specific antibodies. PMID- 11017798 TI - Mapping of the Tobacco mosaic virus movement protein and coat protein subgenomic RNA promoters in vivo. AB - The Tobacco mosaic virus movement protein (MP) and coat protein (CP) are expressed from 3'-coterminal subgenomic RNAs (sgRNAs). The transcription start site of the MP sgRNA, previously mapped to positions 4838 (Y. Watanabe, T. Meshi, and Y. Okada (1984), FEBS Lett. 173, 247-250) and 4828 (K. Lehto, G. L. Grantham, and W. O. Dawson (1990), Virology 174, 145-157) for the TMV OM and U1 strains, respectively, has been reexamined and mapped to position 4838 for strain U1. Sequences of the MP and CP sgRNA promoters were delineated by deletion analysis. The boundaries for minimal and full MP sgRNA promoter activity were localized between -35 and +10 and -95 and +40, respectively, relative to the transcription start site. The minimal CP sgRNA promoter was mapped between -69 and +12, whereas the boundaries of the fully active promoter were between -157 and +54. Computer analysis predicted two stem-loop structures (SL1 and SL2) upstream of the MP sgRNA transcription start site. Deletion analysis and site-directed mutagenesis suggested that SL1 secondary structure, but not its sequence, was required for MP sgRNA promoter activity, whereas a 39-nt deletion removing most of the SL2 region increased MP sgRNA accumulation fourfold. Computer-predicted folding of the fully active CP sgRNA promoter revealed one long stem-loop structure. Deletion analysis suggested that the upper part of this stem-loop, located upstream of the transcription start site, was essential for transcription and that the lower part of the stem had an enhancing role. PMID- 11017799 TI - Identification and analysis of vaccinia virus palmitylproteins. AB - Vaccinia virus encodes at least eight proteins that incorporate label from tritiated palmitic acid when it is added to infected cell cultures. Three of these palmitylproteins are encoded by the A33R, B5R, and F13L open reading frames and migrate by gel electrophoresis with relative molecular masses of 23-28, 42, and 37 kDa, respectively. In this report we provide evidence that the A22R and A36R open reading frames also encode palmitylproteins with apparent molecular masses of 22 and 50-55 kDa, respectively. Furthermore, the hemagglutinin protein (A56R) from the Copenhagen strain is shown to be palmitylated while the hemagglutinin protein from the WR and IHD-J strains is not. A 94-kDa VV palmitylprotein appears to be a multimeric complex composed of the B5R protein and possibly others. All vaccinia-encoded palmitylproteins are present in the membranous fraction of cells and are specific for the trans-Golgi network membrane-enveloped forms of the virus, suggesting that these proteins play a role in the envelopment and egress of virions or the infectivity of released virus. PMID- 11017800 TI - Molecular studies on bromovirus capsid protein. VII. Selective packaging on BMV RNA4 by specific N-terminal arginine residuals. AB - An arginine-rich RNA-binding motif (ARM) found at the N-proximal region of Brome mosaic virus (BMV) coat protein (CP) adopts alpha-helical conformation and shares homology with CPs of plant and insect RNA viruses, HIV-Rev and Tat proteins, bacterial antiterminators, and ribosomal splicing factors. The ARM of BMV CP, consisting of amino acids 9 through 21 with six arginine residues, is essential for RNA binding and subsequent packaging. In this study analysis of the alpha helical contents of wild-type and mutant peptides by circular dichroism spectra identified protein determinants required for such conformation. Electrophoretic mobility-shift assays between viral RNA and BMV CP peptides with either proline or alanine substitutions revealed that the interaction is nonspecific. Expression in vivo of mature full-length BMV CP subunits, having the same substitutions for each arginine within the ARM, derived from biologically active clones was found to be competent to assemble into infectious virions and cause visible symptom phenotypes in whole plants. However, analysis of virion progeny RNA profiles of CP variants and subsequent in vitro reassembly assays between mutant CP and four BMV RNAs unveiled the ability of arginine residues at positions 10, 13, or 14 of the ARM to confer selective packaging of BMV RNA4. Thus, BMV CP contains determinants that specifically interact with RNA4 to ensure selective packaging. PMID- 11017801 TI - Characterization of phi 13, a bacteriophage related to phi 6 and containing three dsRNA genomic segments. AB - The three dsRNA genomic segments of bacteriophage Phi 13 were copied as cDNA and the nucleotide sequences were determined. The organization of the genome is similar to that of Phi 6, and there is significant similarity in the amino acid sequences of the proteins of the polymerase complex and one of the membrane proteins, P6. There is little or no similarity in the nucleotide sequences. Several features of the viral proteins differ markedly from those of Phi 6. Although both phages are covered by a lipid-containing membrane, the protein compositions are different. The host attachment protein consists of two peptides rather than one and the phage attaches directly to the LPS of the host rather than to a Type IV pilus. Despite the differences in the structure of the membranes, the two viruses can successfully exchange the genes for host attachment proteins and thereby change their host specificities. PMID- 11017802 TI - Identification of MEFV-independent modifying genetic factors for familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is a recessively inherited disorder predisposing to renal amyloidosis and associated with mutations in MEFV, a gene encoding a protein of unknown function. Differences in clinical expression have been attributed to MEFV-allelic heterogeneity, with the M694V/M694V genotype associated with a high prevalence of renal amyloidosis. However, the variable risk for patients with identical MEFV mutations to develop this severe complication, prevented by lifelong administration of colchicine, strongly suggests a role for other genetic and/or environmental factors. To overcome the well-known difficulties in the identification of modifying genetic factors, we investigated a relatively homogeneous population sample consisting of 137 Armenian patients with FMF from 127 independent families living in Armenia. We selected the SAA1, SAA2, and APOE genes-encoding serum amyloid proteins and apolipoprotein E, respectively-as well as the patients' sex, as candidate modifiers for renal amyloidosis. A stepwise logistic-regression analysis showed that the SAA1alpha/alpha genotype was associated with a sevenfold increased risk for renal amyloidosis, compared with other SAA1 genotypes (odds ratio [OR] 6. 9; 95% confidence interval [CI] 2.5-19.0). This association, which was present whatever the MEFV genotype, was extremely marked in patients homozygous for M694V (11/11). The risk for male patients of developing renal amyloidosis was fourfold higher than that for female patients (OR=4.0; 95% CI=1.5-10.8). This association, particularly marked in patients who were not homozygous for M694V (34.0% vs. 11.6%), was independent of SAA1-allelic variations. Polymorphisms in the SAA2 or APOE gene did not appear to influence susceptibility to renal amyloidosis. Overall, these data, which provide new insights into the pathophysiology of FMF, demonstrate that susceptibility to renal amyloidosis in this Mendelian disorder is influenced by at least two MEFV-independent factors of genetic origin-SAA1 and sex-that act independently of each other. PMID- 11017803 TI - High frequency of alkaptonuria in Slovakia: evidence for the appearance of multiple mutations in HGO involving different mutational hot spots. AB - Alkaptonuria (AKU) is an autosomal recessive disorder caused by the deficiency of homogentisate 1,2 dioxygenase (HGO) activity. AKU shows a very low prevalence (1:100,000-250,000) in most ethnic groups. One notable exception is in Slovakia, where the incidence of AKU rises to 1:19,000. This high incidence is difficult to explain by a classical founder effect, because as many as 10 different AKU mutations have been identified in this relatively small country. We have determined the allelic associations of 11 HGO intragenic polymorphisms for 44 AKU chromosomes from 20 Slovak pedigrees. These data were compared to the HGO haplotype data available in our laboratory for >80 AKU chromosomes from different European and non-European countries. The results show that common European AKU chromosomes have had only a marginal contribution to the Slovak AKU gene pool. Six of the ten Slovak AKU mutations, including the prevalent G152fs, G161R, G270R, and P370fs mutations, most likely originated in Slovakia. Data available for 17 Slovak AKU pedigrees indicate that most of the AKU chromosomes have their origins in a single very small region in the Carpathian mountains, in the northwestern part of the country. Since all six Slovak AKU mutations are associated with HGO mutational hot spots, we suggest that an increased mutation rate at the HGO gene is responsible for the clustering of AKU mutations in such a small geographical region. PMID- 11017804 TI - Mutation in the gene for connexin 30.3 in a family with erythrokeratodermia variabilis. AB - Erythrokeratodermia variabilis (EKV) is an autosomal dominant keratinization disorder characterized by migratory erythematous lesions and fixed keratotic plaques. All families with EKV show mapping to chromosome 1p34-p35, and mutations in the gene for connexin 31 (Cx31) have been reported in some but not all families. We studied eight affected and three healthy subjects in an Israeli family, of Kurdish origin, with EKV. After having mapped the disorder to chromosome 1p34-p35, we found no mutations in the genes for Cx31, Cx31.1, and Cx37. Further investigation revealed a heterozygous T-->C transition leading to the missense mutation (F137L) in the human gene for Cx30.3 that colocalizes on chromosome 1p34-p35. This nucleotide change cosegregated with the disease and was not found in 200 alleles from normal individuals. This mutation concerns a highly conserved phenylalanine, in the third transmembrane region of the Cx30.3 molecule, known to be implicated in the wall formation of the gap-junction pore. Our results show that mutations in the gene for Cx30.3 can be causally involved in EKV and point to genetic heterogeneity of this disorder. Furthermore, we suggest that our family presents a new type of EKV because of the hitherto unreported association with erythema gyratum repens. PMID- 11017805 TI - A novel mutation of desert hedgehog in a patient with 46,XY partial gonadal dysgenesis accompanied by minifascicular neuropathy. AB - We describe a patient with 46,XY partial gonadal dysgenesis (PGD) who presented with polyneuropathy. Sural nerve pathology revealed peculiar findings characterized by extensive minifascicular formation within the endoneurium and with a decreased density of myelinated fibers. We found, in the patient, a homozygous missense mutation (ATG-->ACG) at the initiating codon in exon 1 of the desert hedgehog (DHH) gene, which predicts a failure of translation of the gene. The same heterozygous mutation was found in the patient's father. This is the first report of a human DHH gene mutation, and the findings demonstrate that mutation of the DHH gene may cause 46, XY PGD associated with minifascicular neuropathy. PMID- 11017807 TI - Silver/Copper ionization is effective for controlof legionella PMID- 11017806 TI - Haploinsufficiency of ALX4 as a potential cause of parietal foramina in the 11p11.2 contiguous gene-deletion syndrome. AB - Heterozygous mutations in MSX2 are responsible for an autosomal dominant form of parietal foramina (PFM). PFM are oval defects of the parietal bones that are also a characteristic feature of a contiguous gene-deletion syndrome caused by a proximal deletion in the short arm of chromosome 11 (Potocki-Shaffer syndrome). We have identified a human bacterial artificial chromosome (BAC) clone mapping to chromosome 11, containing a region homologous to the human homeobox gene MSX2. Further sequence analysis demonstrated that the human orthologue (ALX4) of the mouse Aristaless-like 4 gene (Alx4) is contained within this 11p clone. We used FISH to test for the presence-or for the heterozygous deletion-of this clone in two patients with the 11p11.2-deletion syndrome and showed that this clone is deleted in these patients. ALX4 and Alx4 were shown to be expressed in bone and to be absent from all other tissues tested. The involvement of Alx4 in murine skull development, its bone-specific expression pattern, the fact that Alx4 is a dosage-sensitive gene in mice, and the localization of a human genomic clone containing ALX4 to 11p11.2, with hemizygosity in patients with deletion of 11p11.2 who have biparietal foramina, support the contention that ALX4 is a candidate gene for the PFM in the 11p11.2-deletion syndrome. PMID- 11017808 TI - Practice guidelines for the treatment of tuberculosis. PMID- 11017809 TI - Practice guideline for evaluation of fever and infection in long-term care facilities. PMID- 11017810 TI - Risk factor analysis for breast cellulitis complicating breast conservation therapy. AB - Women who undergo breast conservation therapy for early-stage breast cancer can develop breast cellulitis, a complication for which risk factors are undefined. A matched case-control investigation was conducted to identify risk factors for the development of breast cellulitis among patients who have undergone breast conservation therapy. Patients comprised 17 patients with cases of breast cellulitis diagnosed after partial mastectomies that had been performed from 1992 through 1997 and 34 control patients who were matched to case-patients by date of breast lumpectomy and by primary surgeon. Statistical analyses indicated the following factors were associated with breast cellulitis: drainage of a hematoma (P=.010); postoperative ecchymosis (P=.021); lymphedema (odds ratio [OR], 10. 154; 95% confidence interval [CI], 1.348-208.860); resected breast tissue volume (OR, 1.456; 95% CI, 1.035-2.168); and previous number of breast seroma aspirations (OR, 3.445; 95% CI, 1.036-19.771). This is the first matched case control study to identify risk factors for the development of breast cellulitis after breast conservation therapy. PMID- 11017811 TI - Corticosteroid treatment of eosinophilic meningitis. AB - The role of corticosteroids in the treatment of eosinophilic meningitis has not been definitely established. Patients given a 2-week course of prednisolone (treatment group), 60 mg/day, were compared with those given placebo (control group) in a randomized, double-blind trial. Fifty-five patients were enrolled in each group. There were significant differences between the treatment and control groups, with regard to the number of patients who still had headache after 14 days (5 vs. 25, respectively; P=.00004), the median length of time until complete disappearance of headache (5 vs. 13 days, respectively; P=.00000), and the number of patients who had repeat lumbar puncture (7 vs. 22, respectively; P=.002). Serious side effects were not detected. These results indicate that a 2-week course of prednisolone was beneficial in relieving headache in patients with eosinophilic meningitis. PMID- 11017812 TI - Adult adenovirus infections: loss of orphaned vaccines precipitates military respiratory disease epidemics. For the Adenovirus Surveillance Group. AB - Adenovirus vaccines have greatly reduced military respiratory disease morbidity since the 1970s. However, in 1995, for economic reasons, the sole manufacturer of these vaccines ceased production. A population-based adenovirus surveillance was established among trainees with acute respiratory illness at 4 US military training centers as the last stores of vaccines were depleted. From October 1996 to June 1998, 1814 (53.1%) of 3413 throat cultures for symptomatic trainees (78% men) yielded adenovirus. Adenovirus types 4, 7, 3, and 21 accounted for 57%, 25%, 9%, and 7% of the isolates, respectively. Unvaccinated trainees were much more likely than vaccinated trainees to be positive for types 4 or 7 (odds ratio [OR] = 28.1; 95% CI, 20.2-39.2). Two training centers experienced epidemics of respiratory disease affecting thousands of trainees when vaccines were not available. Until a new manufacturer is identified, the loss of orphaned adenovirus vaccines will result in thousands of additional preventable adenovirus infections. PMID- 11017813 TI - A tale of two vaccines. PMID- 11017814 TI - Pseudallescheria boydii brain abscess successfully treated with voriconazole and surgical drainage: case report and literature review of central nervous system pseudallescheriasis. AB - Pseudallescheria boydii and its asexual form, Scedosporium apiospermum, are ubiquitous, saprophytic fungi that commonly cause cutaneous infection. However, in certain circumstances, P. boydii can also cause invasive disease, which can involve the central nervous system (CNS). When the CNS becomes involved, treatment is difficult, therapeutic options are limited, and the prognosis is poor. We report a case of Pseudallescheria brain abscess successfully treated with surgical drainage and systemic voriconazole, the first such case to be described in the literature. We also review previously reported cases of CNS pseudallescheriasis and evaluate therapeutic options. PMID- 11017815 TI - Deaths of children during an outbreak of hand, foot, and mouth disease in sarawak, malaysia: clinical and pathological characteristics of the disease. For the Outbreak Study Group. AB - From April through June 1997, 29 previously healthy children aged <6 years (median, 1.5 years) in Sarawak, Malaysia, died of rapidly progressive cardiorespiratory failure during an outbreak of hand, foot, and mouth disease caused primarily by enterovirus 71 (EV71). The case children were hospitalized after a short illness (median duration, 2 days) that usually included fever (in 100% of case children), oral ulcers (66%), and extremity rashes (62%). The illness rapidly progressed to include seizures (28%), flaccid limb weakness (17%), or cardiopulmonary symptoms (of 24 children, 17 had chest radiographs showing pulmonary edema, and 24 had echocardiograms showing left ventricular dysfunction), resulting in cardiopulmonary arrest soon after hospitalization (median time, 9 h). Cardiac tissue from 10 patients showed normal myocardium, but central nervous system tissue from 5 patients showed inflammatory changes. Brain stem specimens from 2 patients were available, and both specimens showed extensive neuronal degeneration, inflammation, and necrosis, suggesting that a central nervous system infection was responsible for the disease, with the cardiopulmonary dysfunction being neurogenic in origin. EV71 and possibly an adenovirus, other enteroviruses, or unknown cofactors are likely responsible for this rapidly fatal disease. PMID- 11017816 TI - Analysis of 42 cases of septicemia caused by an epidemic strain of methicillin resistant Staphylococcus aureus: evidence of resistance to vancomycin. AB - Recent case reports of vancomycin treatment failures in the United States, Japan, and France have prompted a retrospective analysis of 42 cases of septicemia caused by epidemic methicillin-resistant Staphylococcus aureus strain 15 (EMRSA 15), which is the most prevalent epidemic strain of methicillin-resistant S. aureus in the United Kingdom; all cases occurred in a teaching hospital in Manchester, United Kingdom, between 1994 and 1998. Mortality was lowest (4%) in patients with rifampin-susceptible isolates treated with vancomycin and rifampin. It rose to 38% in patients who were treated with both antibiotics but in whom the organism became resistant to rifampin during therapy, and it reached 78% in patients who had rifampin-resistant isolates or in whom rifampin was contraindicated (P<.0001; Fisher exact test, 2-tailed). All isolates were susceptible to vancomycin by conventional laboratory testing, but susceptibility was lost by growth in vancomycin in vitro, becoming resistant at a minimum inhibitory concentration of 8 mg/L. This was associated with accumulation of cell wall material. The deoxyribonucleic acid fingerprint remained unchanged. This study suggests that rifampin played a key role in the prevention of deaths caused by an epidemic strain of methicillin-resistant S. aureus that readily gave rise to a subpopulation with reduced susceptibility to vancomycin. PMID- 11017817 TI - Nosocomial bloodstream infections caused by Acinetobacter species in United States hospitals: clinical features, molecular epidemiology, and antimicrobial susceptibility. AB - We examined the clinical and epidemiological features of nosocomial bloodstream infections (BSIs) caused by Acinetobacter species and observed from 1 March 1995 through 28 February 1998 at 49 United States hospitals (SCOPE National Surveillance Program). Acinetobacter species were found in 24 hospitals (49%) and accounted for 1.5% of all nosocomial BSIs reported. One hundred twenty-nine isolates were identified either as A. baumannii (n=111) or other Acinetobacter species (n=18). Patients with A. baumannii BSI, compared with patients with nosocomial BSI caused by other gram-negative pathogens, were more frequently observed in the intensive care unit (69% vs. 47%, respectively; P<.001; odds ratio [OR] 2.4; 95% confidence interval [CI] 1.6-3.7) and were more frequently receiving mechanical ventilation (58% vs. 30%, respectively; P<.001; OR 3.2; 95% CI 2.1-4.8). Crude mortality in patients with A. baumannii BSI was 32%. Molecular relatedness of strains was studied by use of polymerase chain reaction-based fingerprinting. Clonal spread of a single strain occurred in 5 hospitals. Interhospital spread of epidemic A. baumannii strains was not observed. The most active antimicrobial agents against A. baumannii (90% minimum inhibitory concentration values) were imipenem (1 mg/L; 100% of isolates susceptible), amikacin (8 mg/L; 96%), tobramycin (4 mg/L; 92%), and doxycycline (4 mg/L; 91%). Thirty percent of isolates were resistant to > or =4 classes of antimicrobials and were considered to be multidrug resistant. PMID- 11017818 TI - Hospital admissions for rotavirus infection in the Netherlands. AB - The development of a vaccine against rotavirus (RV) infection has necessitated the estimation of the number of hospitalizations for RV infection in the Netherlands. During 1998, pediatricians have reported all hospitalizations with RV infection and supplied information on the duration of admission, clinical picture, indication for admission, and treatment. Also, data from the National Disease Registry on hospitalizations for gastroenteritis (International Classification of Disease codes 006.6. 006.8, 009, and 558.9) and laboratory surveillance data for 1996-1998 were combined in a linear regression model to indirectly estimate the incidence and proportion of hospitalizations attributable to RV infection. The direct estimate of admissions for RV infection in children aged <5 years was 0.9 per 1000, and the indirect estimate was 2.7 per 1000 in 1998 (1996, 3.4; 1997, 1.6). The proportion of hospitalizations for gastroenteritis attributable to RV ranged from 32% in 1997 to 58% in 1996. PMID- 11017820 TI - High rate of candidemia in patients sustaining injuries in a bomb blast at a marketplace: a possible environmental source. AB - In this study, a cluster of candidemia among patients sustaining injuries in a bomb blast at a marketplace was investigated by means of a multivariate analysis, a case-control study, and quantitative air sampling. Candidemia occurred in 7 (30%) of 21 patients (58% of those admitted to the intensive care unit [ICU]) between 4 and 16 days (mean, 12 days) after the injury and was the single most frequent cause of bloodstream infections. Inhalation injury was the strongest predictor for candidemia by multivariate analysis. Candidemia among the case patients occurred at a significantly higher rate than among comparable trauma patients injured in different urban settings, including a pedestrian mall (2 of 29; P=. 02), and among contemporary ICU control patients (1 of 40; P=.001). Air sampling revealed exclusive detection of Candida species and increased mold concentration in the market in comparison with the mall environment. These findings suggest a role for an exogenous, environmental source in the development of candidemia in some trauma patients. PMID- 11017819 TI - Bacteremia due to Stenotrophomonas maltophilia in patients with hematologic malignancies. AB - Predisposing factors, clinical characteristics, and antimicrobial treatment of 37 hematology patients with Stenotrophomonas maltophilia bacteremia who were seen at the department of hematology of the University La Sapienza (Rome) from 1987 to 1996 were evaluated. The results were compared with a control group of patients with Pseudomonas aeruginosa bacteremia. Profound neutropenia was more prolonged in the S. maltophilia group (P=.025), severe cellulitis occurred only in S. maltophilia-infected patients (11 [30%]; P=.0002), and the bacteremia presented as breakthrough infection in 56% of the cases due to S. maltophilia (vs. only 24% of those due to P. aeruginosa; P=.002). Acute mortality rates associated with S. maltophilia and P. aeruginosa bacteremia were 24% and 21%, respectively. In both groups, profound neutropenia and hypotension at the onset of bacteremia, duration of profound neutropenia during bacteremia, severity-of-illness score > or =4, and inappropriate antibacterial treatment were factors significantly associated with death. Most S. maltophilia isolates were resistant to aminoglycosides, beta lactams, and ciprofloxacin. Cotrimoxazole and ticarcillin-clavulanic acid showed borderline activity. Prompt administration of in vitro-active antibiotics may improve the prognosis of S. maltophilia bacteremia, especially for immunocompromised patients, and novel drug combinations are needed for the treatment of severe infections. PMID- 11017821 TI - The role of physical proximity in nosocomial diarrhea. AB - To examine physical proximity as a risk factor for the nosocomial acquisition of Clostridium difficile-associated diarrhea (CDAD) and of antibiotic-associated diarrhea (AAD), we assessed a retrospective cohort of 2859 patients admitted to a community hospital from 1 March 1987 through 31 August 1987. Of these patients, 68 had nosocomial CDAD and 54 had nosocomial AAD. In multivariate analysis, physical proximity to a patient with CDAD (relative risk [RR], 1.86; 95% confidence interval [CI], 1.06-3.28), exposure to clindamycin (RR, 4.22; 95% CI, 2.11-8.45), and the number of antibiotics taken (RR, 1.49; 95% CI, 1.23-1.81) were significant. For patients with nosocomial AAD, exposure to a roommate with AAD (RR, 3.94; 95% CI, 1. 27-12.24), a stay in an intensive care unit or cardiac care unit (RR, 1.93; 95% CI, 1.05-3.53), and the number of antibiotics taken (RR, 2.01; 95% CI, 1.67-2.40) were significant risk factors. Physical proximity may be an independent risk factor for acquisition of nosocomial CDAD and AAD. PMID- 11017822 TI - Adults with meningitis caused by oxacillin-resistant Staphylococcus aureus. AB - Since 1995, 11 adult patients with oxacillin-resistant Staphylococcus aureus (ORSA) meningitis have been identified at Chang Gung Memorial Hospital-Kaohsiung, in Kaohsiung, Taiwan. The 11 patients were 8 men and 3 women, aged 17-78 years. A postneurosurgical state was an underlying condition for all, and fever and disturbances in consciousness were the most common clinical manifestations. Infection with S. aureus only was found in 8 patients, and mixed infection was found in the other 3. The 8 patients with meningitis caused by S. aureus only were mainly treated with intravenous vancomycin, 2-4 g/day; 4 of these patients died. Although ORSA meningitis is uncommon among adults with culture-proven bacterial meningitis, its incidence has been increasing in recent years. The diagnosis of adult ORSA meningitis can be confirmed only with a positive culture of cerebrospinal fluid, and the choice of initial empirical antibiotics must be guided by the accumulated data concerning the relative frequency of the implicated pathogens found at each institution. Despite the high rate of mortality associated with ORSA meningitis, intravenous vancomycin therapy seems to be one of the best choices for management of this condition in adults. PMID- 11017823 TI - Application of a geographic information system to the tracking and control of an outbreak of shigellosis. AB - A personal computer-based commercial geographic information system (GIS) was applied to an outbreak of Shigella sonnei infection at Fort Bragg, North Carolina. We used a database consisting of demographic, temporal, and home address information for all recognized cases of S. sonnei that occurred among health care beneficiaries from 23 May 1997 through 14 August 1997. We imported this database into the GIS, which contained a digitized basemap of the local community. Through simultaneous examination of temporal and spatial distribution of the 59 identified cases of S. sonnei, a focus of infection in a single housing area was identified. Targeted education among residents of the neighborhood in which there was intense transmission was associated with prompt extinction of the epidemic. A GIS offers an efficient and practical way to directly visualize the dynamics of transmission of infectious diseases in the setting of a community outbreak. PMID- 11017824 TI - Diagnostic virology. AB - Diagnostic virology has now entered the mainstream of medical practice. Multiple methods are used for the laboratory diagnosis of viral infections, including viral culture, antigen detection, nucleic acid detection, and serology. The role of culture is diminishing as new immunologic and molecular tests are developed that provide more rapid results and are able to detect a larger number of viruses. This review provides specific recommendations for the diagnostic approach to clinically important viral infections. PMID- 11017825 TI - Infection control in long-term care facilities. AB - Infections are common in long-term care facilities. The most frequent endemic infections are urinary infection, respiratory infection, and skin and soft tissue infections. Outbreaks also occur frequently, and some facilities have a high prevalence of colonization of residents with antimicrobial-resistant organisms. Our understanding of infections and the development of infection-control programs for long-term care facilities have progressed greatly over the past 15 years. Whereas the occurrence of infections has been described and specific guidelines for infection-control programs in long-term care facilities have been developed, there is still limited evaluation of the effectiveness of programs or specific interventions to support prioritization of infection-control resources. In addition, the spectrum of patients and care delivered in long-term care facilities continues to evolve. Increasingly, chronic care patients, including those requiring chronic respirator therapy, dialysis, or percutaneous feeding tubes, are cared for in these facilities. Our understanding of prevention of infection in these patients remains limited. Important questions include what interventions may prevent endemic infections, what are the most effective means to identify outbreaks early, and what interventions may minimize the prevalence of antimicrobial-resistant organisms. Programs to optimize antimicrobial use need to be developed. Thus, although progress in understanding and practice has been made, important questions remain. PMID- 11017826 TI - Urinary tract infection in long-term-care facility residents. AB - Urinary tract infection is the most frequent bacterial infection in residents of long-term-care facilities. Most infections are asymptomatic, with a remarkable prevalence of asymptomatic bacteriuria of 15%-50% among all residents. The major reasons for this high prevalence are chronic comorbid illnesses with neurogenic bladder and interventions to manage incontinence. Prospective, randomized, comparative trials of therapy and no therapy for asymptomatic bacteriuria among nursing home residents have repeatedly documented that antimicrobial treatment had no benefits. However, there is substantial diagnostic uncertainty in determining whether an individual with a positive urine culture has symptomatic or asymptomatic infection when there is clinical deterioration and there are no localized findings. In the noncatheterized resident, urinary infection is an infrequent source of fever but may not be definitively excluded. The use of antimicrobials for treatment of urinary infection is part of the larger concern about appropriate antimicrobial use in long-term-care facilities and the impacts of the selective pressure of antimicrobials on colonization and infection with resistant organisms. PMID- 11017827 TI - Bacterial monopolists: the bundling and dissemination of antimicrobial resistance genes in gram-positive bacteria. AB - Antibiotic resistance is the unavoidable result of our placing selective pressure on the microbial community. Advances in molecular biology techniques in the past 2 decades have allowed us to greatly improve our understanding of the mechanisms by which resistance emerges and disseminates among human pathogenic bacteria. Gram-positive bacteria employ a diverse array of elements, including plasmids, transposons, insertion sequences, and bacteriophages, to disseminate resistance. An understanding of these mechanisms and their prevalence can improve our ability to treat clinical infections in hospitalized patients, as well as to predict and control the spread of resistant bacteria in the nosocomial environment. PMID- 11017828 TI - Foodborne listeriosis. AB - Listeria monocytogenes emerged as an important foodborne pathogen in the latter part of the 20th century. Clinical syndromes caused by this microorganism include sepsis in the immunocompromised patient, meningoencephalitis in infants and adults, and febrile gastroenteritis. Focal infections at other sites are less frequent. Listeria species are commonly found in raw and unprocessed food products. Major outbreaks of listeriosis, with high morbidity and mortality, have been caused by a variety of foods, including soft cheeses, delicatessen meats, and vegetable products. Improved detection methods, dietary recommendations, and, in some cases, preemptive antibiotic treatment or prophylaxis have reduced the incidence of sporadic listeriosis infections in the United States. Microbial virulence factors distinguishing environmental strains of L. monocytogenes from invasive strains causing foodborne illness and host factors promoting human infection remain incompletely understood. PMID- 11017829 TI - Population mobility and infectious diseases: the diminishing impact of classical infectious diseases and new approaches for the 21st century. AB - In an increasingly globalized world, rapid population mobility and migration is reducing the differences in infectious disease epidemiology between regions of the world. The movement and relocation of populations between locations where the prevalence and incidence of infections are markedly different poses current and future challenges to those involved in clinical infectious diseases and public health program management. Historically, international attention has focused on the screening and treatment of acute infections of epidemic potential, but, as immigration significantly changes the demography of many nations, chronic infections will require increased attention. In countries with large mobile populations, the population-based burden of infections with long latency periods or significant noninfectious sequelae will make up an increasing amount of the infectious disease caseload and will require more-modern approaches than the traditional screening of arrivals. The globalization of chronic infectious disease epidemiology will require corresponding development of integrated programs to anticipate and manage these diseases in response to an increasingly mobile patient population. PMID- 11017830 TI - Cadaver donor screening for infectious agents in solid organ transplantation. AB - The transmission of infection by a cadaver donor organ can result not only in loss of the allograft but also in death of the immunosuppressed recipient. Despite the shortage of cadaver organ donors, every donor must be evaluated thoroughly for the potential transmission of infectious disease, because the consequences of the organ donor events can have a profound effect on the transplant outcome. This review summarizes current knowledge about serological screening of organ donors to determine the suitability of organs from cadaver donors for transplantation. PMID- 11017831 TI - Coronary artery disease and human immunodeficiency virus infection. AB - Recent reports of myocardial infarctions in young persons infected with human immunodeficiency virus (HIV) who are receiving protease inhibitor therapy have raised concerns about premature coronary artery disease in this population. Endothelial dysfunction, hypercoagulability, hypertriglyceridemia, and abnormal coronary artery pathology were in fact associated with HIV infection prior to the availability of protease inhibitor therapy. Newly recognized risk factors, such as insulin resistance, hypercholesterolemia, and fat redistribution syndrome, may exacerbate underlying atherosclerotic risk for patients receiving protease inhibitors. Data on the incidence of myocardial infarction among these patients are largely limited to case reports but are of concern. Pending the availability of further data, it is prudent to monitor these patients for hyperlipidemia and consider interventions to modify cardiac risk factors. PMID- 11017832 TI - Clinical implications of identifying non-B subtypes of human immunodeficiency virus type 1 infection. AB - Although human immunodeficiency virus type 1 (HIV-1) infection in the United States has predominantly involved subtype B, increasing global travel is leading to wider dissemination of genetically heterogeneous subtypes. While physicians depend on HIV-1 viral load measurements to guide antiretroviral therapy, commonly used molecular assays may underestimate the viral load of patients with non-B subtypes. Nine patients with non-B subtypes of HIV-1 were identified by physicians who suspected a non-B subtype on the basis of a low or undetectable HIV-1 viral load, by the Amplicor HIV-1 Monitor test, version 1.0, in conjunction with either a declining CD4 cell count or history of travel outside the United States. Use of version 1.5 of the Amplicor HIV-1 Monitor test detected a median HIV-1 viral load that was 2.0 log(10) RNA copies/mL higher than was determined with version 1.0. Clinical management was altered in all cases after diagnosis of a non-B-subtype infection. These cases demonstrate that it is critical for physicians to suspect and diagnose non-B subtypes of HIV-1 so that an assay with reliable subtype performance can be used to guide antiretroviral therapy. PMID- 11017833 TI - Weight loss and wasting remain common complications in individuals infected with human immunodeficiency virus in the era of highly active antiretroviral therapy. AB - It has been postulated that the use of highly active antiretroviral therapy (HAART) would reduce the occurrence of human immunodeficiency virus (HIV) associated weight loss and wasting. To test this assumption, we evaluated, by means of longitudinal analysis, a prospective cohort of 469 HIV-infected individuals enrolled in a study of the impact of HIV on nutrition. Overall, 156 individuals in the cohort (33.5%) met at least 1 of these definitions of wasting. Furthermore, 58% of the cohort (289 patients) lost >1.5 kg of weight in a 6-month period between any 2 study visits. More than 50% of the cohort was receiving HAART at the time that they met 1 of the definitions of wasting; with regard to the occurrence of wasting; no differences were related to therapy. PMID- 11017834 TI - Dramatic improvement of renal dysfunction in a human immunodeficiency virus infected woman treated with highly active antiretroviral therapy. AB - We report a single case documenting substantial improvement in the course of human immunodeficiency virus (HIV)-associated, biopsy-proven nephropathy after introduction of highly active antiretroviral therapy. Our case report joins several others recording improvement or stabilization in the course of nephropathy following better control of HIV replication. PMID- 11017835 TI - The tolerability of efavirenz after nevirapine-related adverse events. AB - Eight patients who were infected with human immunodeficiency virus, and who had each sustained an adverse drug reaction while following a regimen including nevirapine, were switched to a regimen including efavirenz. None of the patients experienced adverse events identical to that which necessitated discontinuation of nevirapine. This study demonstrates that adverse events related to nevirapine are not a class-specific effect. PMID- 11017836 TI - Human immunodeficiency virus infection, AIDS, and smoking cessation: the time is now. AB - Treatments for persons who are infected with human immunodeficiency virus (HIV) or who have developed AIDS have advanced to the point where death is no longer the inevitable outcome of diagnosis. Combination antiretroviral therapy has made HIV infection less of a terminal condition and more of a medically manageable chronic disease. Thus, efforts to improve the health status and quality of life of HIV-infected persons have become one of the highest treatment priorities for the next decade. Cigarette smoking is highly prevalent among HIV-infected persons, and quitting smoking would greatly improve the health status of these individuals. However, to date, no studies have evaluated the efficacy of a smoking-cessation intervention specifically tailored to this population. This article reviews the evidence and rationale for advancing smoking-cessation treatments specifically tailored to the needs of HIV-infected persons and provides recommendations for future treatment studies. PMID- 11017837 TI - Two cases of human urinary tract infection complicated by hemolytic uremic syndrome caused by verotoxin-producing Escherichia coli. AB - In 1993, 2 cases of urinary tract infection (UTI) caused by verotoxin-producing Escherichia coli were diagnosed at Rigshospitalet in Copenhagen, Denmark. Neither of the patients had any previous history of diarrhea. We suggest that E. coli strains isolated from UTI be examined for the production of verotoxin when hemolytic uremic syndrome is clinically suspected. PMID- 11017838 TI - Mycobacterium thermoresistible recovered from a cutaneous lesion in an otherwise healthy individual. AB - This is the first report of coinfection by Mycobacterium thermoresistible and Mycobacterium fortuitum and only the fifth case of human infection by M. thermoresistible reported in the world literature. PMID- 11017839 TI - Portal vein and bone involvement in disseminated cat-scratch disease: report of 2 cases. AB - Cat-scratch disease (CSD) is a common cause of regional lymphadenopathy. We describe 2 children with an unusual presentation of disseminated CSD, the first one presenting with persistent fever, multilocular abscesses in liver and spleen as well as osteolytic lesions in the lumbar spine and the second one with portal vein thrombosis and severe ascites. PMID- 11017840 TI - Thiabendazole for the treatment of strongyloidiasis in patients with hematologic malignancies. AB - A total of 21 patients with hematologic malignancies were given thiabendazole for treatment of strongyloidiasis. Fifteen patients were cured. Since there were no relapses, it is unlikely that maintenance therapy has a role in the management of strongyloidiasis in this population of patients. PMID- 11017841 TI - The use of adjuvant interferon-gamma therapy for hepatosplenic Blastoschizomyces capitatus infection in a patient with leukemia. AB - Hepatosplenic fungal infections are a devastating complication of neutropenia. Despite aggressive antifungal therapy, clinical response may be poor. We describe a case of hepatosplenic Blastoschizomyces capitatus infection that responded to adjuvant interferon-gamma therapy. PMID- 11017842 TI - Vancomycin-induced neutropenia resolves after substitution with teicoplanin. AB - Neutropenia is an uncommon adverse effect associated with prolonged vancomycin therapy. Neutrophil counts normally recover after discontinuation of vancomycin in this situation, but treatment options are needed for those patients who require ongoing antibiotic therapy. We describe a case of vancomycin-induced neutropenia in which the neutropenia resolved after vancomycin was replaced by the structurally related compound teicoplanin. PMID- 11017843 TI - Treatment of latent tuberculosis infection in patients aged > or =35 years. AB - Treating patients aged > or =35 years for tuberculosis infection has been controversial because of the hepatotoxic effects of isoniazid. A 2-year retrospective cohort study of outpatient medical records determined the completion rate in this age group and identified risk factors associated with isoniazid-associated hepatotoxicity. Isoniazid preventative therapy was well tolerated. However, toxicity occurred in women receiving concomitant medications and men who used alcohol. PMID- 11017844 TI - Isolation and characterization of adenovirus 5 from the brain of an infant with fatal cerebral edema. AB - This report describes a fatal case of cerebral edema caused by adenovirus in a previously healthy 18-month-old infant who developed skin rash, pulmonary congestion, and fever and who died 6 days later. Adenovirus hexon gene sequences were detected in brain tissue and brain tissue cultures. The virus was typed as adenovirus 5. PMID- 11017845 TI - Chemokine concentrations in nasal washings of infants with rhinovirus illnesses. AB - We determined RANTES (regulated on activation, normal T cell expressed and secreted) and interleukin-8 (IL-8) concentrations, and total white blood cell (WBC) and differential counts in nasal wash samples from rhinovirus-infected infants presenting with wheezing or acute upper respiratory illness alone and compared them with those from healthy infants. RANTES concentrations were significantly greater in acute samples from wheezy patients than in those from patients with acute upper respiratory illness only, or in control samples. IL-8 concentrations and WBC and neutrophil counts were significantly greater in acute samples from wheezy infants and patients with upper respiratory illness alone than in control samples, but they did not differ significantly between the 2 patient groups. PMID- 11017846 TI - Veillonella parvula meningitis: case report and review of Veillonella infections. AB - Veillonella parvula is a small, nonfermentative anaerobic gram-negative coccus that is part of the normal flora of the mouth, gastrointestinal tract, and vagina in humans. When isolated from clinical specimens, V. parvula is often regarded as a contaminant or commensal, but it has been implicated as a pathogen in infections of the sinuses, lungs, heart, bone, and central nervous system. Meningitis, however, is extremely rare; to our knowledge, only 2 cases have been previously described in the literature. We report a case of V. parvula meningitis and review the literature on Veillonella infections. PMID- 11017847 TI - Tuberculous aortitis with an aortoduodenal fistula presenting as recurrent gastrointestinal bleeding. AB - Tuberculous aortitis with a tuberculous mycotic aneurysm and an aortoduodenal fistula was diagnosed in a 38-year-old man with tuberculous cervical lymphadentitis and a 3-month history of recurrent gastrointestinal bleeding, in whom extensive investigation of the digestive tract had not revealed a bleeding lesion. Either by septic embolism or by direct extension from a neighboring focus, tuberculous infection can cause a mycotic aortic aneurysm with subsequent fistulation to the duodenum. PMID- 11017848 TI - Intradermal postexposure rabies vaccine regimens. PMID- 11017849 TI - Treatment of acute lung injury attributed to leptospirosis. PMID- 11017850 TI - Infection may not have been caused by Exophiala jeanselmei. PMID- 11017851 TI - Detection of selected fastidious bacteria. PMID- 11017852 TI - Pathogenesis of right-side endocarditis. PMID- 11017853 TI - Hepatitis A virus vaccination among injecting drug users: Do We have to change the vaccination schedule? PMID- 11017854 TI - Possibility of the use of oral long-acting tetracyclines in the treatment of Lyme neuroborreliosis. PMID- 11017856 TI - Mycotic aneurysms of the aorta caused by infection with Pasteurella multocida. AB - We evaluated a patient for mycotic aneurysms caused by Pasteurella multocida. We treated Pasteurella aortitis medically with ciprofloxacin, and the patient has had long-term survival. PMID- 11017857 TI - Is a second urine specimen necessary for the diagnosis of asymptomatic bacteriuria? AB - By use of pulse-field gel electrophoresis, we evaluated the molecular identity of 32 Escherichia coli isolates obtained in 2 consecutive urine cultures from 16 patients as part of a large study of asymptomatic bacteriuria in diabetic women and found different E. coli isolates in 7 of 16 patients, meaning that nearly half (44%) of the patients who had been previously classified as having asymptomatic bacteriuria were reinfected with a different strain. PMID- 11017858 TI - Strongyloides-infected patients at Charleston area medical center, West Virginia, 1997-1998. AB - Twenty Strongyloides-infected patients were diagnosed at West Virginia's Charleston Area Medical Center in 1997 and 1998. We recommend that strongyloidiasis, which can be fatal, be a reportable disease in West Virginia. PMID- 11017859 TI - Yellow fever vaccination of human immunodeficiency virus-infected patients: report of 2 cases. AB - Yellow fever vaccine (17D, a live attenuated virus vaccine) was effective and safe in 2 human immunodeficiency virus-infected patients without severe immunosuppression, one of whom traveled to Kenya and the other of whom traveled to Senegal. PMID- 11017860 TI - Introduction PMID- 11017861 TI - The impact of changing pathogens of serious infections in hospitalized patients. AB - New epidemiological patterns are being observed for multidrug-resistant nosocomial organisms. Current problems include the appearance of resistance determinants in organisms that are virulent enough to cause infection in patients with normal host defenses. In addition, multidrug-resistant organisms are spreading from health care to community settings, and organisms from the community are spreading to health care settings. The appearance and spread of resistance can be examined both at a molecular level and on a larger scale involving several pathways. Potential pathways within institutions include the following: introduction of new strains from outside sources (e.g., patients or health care workers from other institutions); exchange of resistance determinants via genetic mutation or transfer of genetic material; emergence or selection of resistant strains following exposure to antimicrobials; and clonal dissemination. Strategies such as multidisciplinary management of infections, appropriate infection control measures, and surveillance of resistance patterns are necessary to address the problem of resistance. PMID- 11017862 TI - Inadequate antimicrobial treatment: an important determinant of outcome for hospitalized patients. AB - Inadequate antimicrobial treatment, generally defined as microbiological documentation of an infection that is not being effectively treated, is an important factor in the emergence of infections due to antibiotic-resistant bacteria. Factors that contribute to inadequate antimicrobial treatment of hospitalized patients include prior antibiotic exposure, use of broad-spectrum antibiotics, prolonged length of stay, prolonged mechanical ventilation, and presence of invasive devices. Strategies to minimize inadequate treatment include consulting an infectious disease specialist, using antibiotic practice guidelines, and identifying quicker methods of microbiological identification. In addition, clinicians should determine the prevailing pathogens that account for the community-acquired and nosocomial infections identified in their hospitals. Clinicians can improve antimicrobial treatment by using empirical combination antibiotic therapy based on individual patient characteristics and the predominant bacterial flora and their antibiotic susceptibility profiles. This broad-spectrum therapy can then be narrowed when initial culture results are received. Further study evaluating the use of antibiotic practice guidelines and strategies to reduce inadequate treatment is necessary to determine their impact on patient outcomes. PMID- 11017863 TI - Nosocomial bloodstream infections: organisms, risk factors, and implications. AB - In the last 30 years, the frequency, etiology, and epidemiology of bloodstream infections (BSIs) have changed with the evolution of medical care, particularly among the increasing number of hospitalized patients who require intensive care. Although gram-negative bacilli were the predominant nosocomial pathogens in the 1970s, gram-positive cocci have emerged as a more frequent cause of nosocomial BSIs during the 1980s and 1990s. Many gram-positive cocci associated with nosocomial BSIs are now resistant to commonly used antibiotics. Currently, the 3 most common causes of nosocomial BSIs in the United States are coagulase-negative staphylococci, Staphylococcus aureus, and enterococci. The emergence of vancomycin-resistant staphylococcal infections is of particular concern. In addition, the incidence of methicillin-resistant S. aureus (MRSA) infections appears to be increasing; however, the effect of MRSA infection on mortality in hospitalized patients remains unclear. Therefore, newer, more effective antimicrobial therapies are needed to treat BSIs caused by gram-positive cocci are needed. PMID- 11017864 TI - Emerging therapies for serious gram-positive bacterial infections: a focus on linezolid. AB - Respiratory tract infections and skin and soft-tissue infections frequently are caused by gram-positive cocci, and treating these infections with standard antibiotics has recently become problematic. Many of the primary pathogens causing these infections are now resistant to current standard treatment regimens. In addition, the frequency of these infections is increasing, particularly among patients with complex medical conditions. Thus, new and effective antimicrobial agents are needed, and many are currently in various stages of development. Linezolid, the first approved oxazolidinone, has enhanced activity against gram-positive organisms. Recent results of 5 large, randomized, phase 3 trials evaluating linezolid for the treatment of community-acquired pneumonia, nosocomial pneumonia, and uncomplicated and complicated skin and soft tissue infections are encouraging and indicate that linezolid is as effective as standard comparator agents as therapy for these infections. Thus, the recent availability of linezolid offers clinicians a promising new agent for the treatment of serious gram-positive bacterial infections. PMID- 11017865 TI - Strategies for the successful treatment of gram-positive bacterial infections. PMID- 11017867 TI - The Wnt signalling pathway. PMID- 11017868 TI - Flower development pathways. PMID- 11017871 TI - Visualizing myosin's power stroke in muscle contraction. AB - The long-standing swinging crossbridge or lever arm hypothesis for the motor action of myosin heads finds support in recent results from 3-D tomograms of insect flight muscle (IFM) fast frozen during active contraction and from both fluorescence polarization and X-ray diffraction during rapid stretches or releases of isometrically contracting fibers. The latter provide direct evidence for lever arm movements synchronous with force changes. Rebuilding the atomic model of nucleotide-free subfragment 1 (S1) to fit fast-frozen, active IFM crossbridges suggests a two-stage power stroke in which the catalytic domain rolls on actin from weak to strong binding; this is followed by a 5-nm lever arm swing of the light chain domain, which gives a total interaction distance of approx. 12 nm. Comparison of S1 crystal structures with in situ myosin heads suggests that actin binding may be necessary in order to view the full repertoire of myosin motor action. The differing positions of the catalytic domains of actin attached myosin heads in contracting IFM suggest that both the actin-myosin binding energy and the hydrolysis of ATP may be used to cock the crossbridge and drive the power stroke. PMID- 11017872 TI - Integrin cytoplasmic domain-binding proteins. AB - Integrins are a large family of cell surface receptors that mediate cell adhesion and influence migration, signal transduction, and gene expression. The cytoplasmic domains of integrins play a pivotal role in these integrin-mediated cellular functions. Through interaction with the cytoskeleton, signaling molecules, and other cellular proteins, integrin cytoplasmic domains transduce signals from both the outside and inside of the cell and regulate integrin mediated biological functions. Identification and functional analyses of integrin cytoplasmic domain-binding proteins have been pursued intensively. In recent years, more cellular proteins have been reported to directly interact with integrin cytoplasmic domains and some of these interactions may play important roles in integrin-mediated biological responses. Integrin (&bgr;) chains, for example, interact with actin-binding proteins (e.g. talin and filamin), which form mechanical links to the cytoskeleton. These and other proteins (e.g. FAK, ILK and novel proteins such as TAP20) might also link integrins to signaling mechanisms and, in some cases (e.g. JAB1) mediate integrin-dependent gene regulation. PMID- 11017873 TI - Use of phosphosynapsin I-specific antibodies for image analysis of signal transduction in single nerve terminals. AB - We have developed a semi-quantitative method for indirectly revealing variations in the concentration of second messengers (Ca(2+), cyclic AMP) in single presynaptic boutons by detecting the phosphorylation of the synapsins, excellent nerve terminal substrates for cyclic AMP- and Ca(2+)/calmodulin-dependent protein kinases. For this purpose, we employed polyclonal, antipeptide antibodies recognising exclusively synapsin I phosphorylated by Ca(2+)/calmodulin-dependent protein kinase II (at site 3) or synapsins I/II phosphorylated by either cAMP dependent protein kinase or Ca(2+)/calmodulin-dependent protein kinase I (at site 1). Cerebellar granular neurones in culture were double-labelled with a monoclonal antibody to synapsins I/II and either of the polyclonal antibodies. Digitised images were analysed to determine the relative phosphorylation stoichiometry at each individual nerve terminal. We have found that: (i) under basal conditions, phosphorylation of site 3 was undetectable, whereas site 1 exhibited some degree of constitutive phosphorylation; (ii) depolarisation in the presence of extracellular Ca(2+) was followed by a selective and widespread increase in site 3 phosphorylation, although the relative phosphorylation stoichiometry varied among individual terminals; and (iii) phosphorylation of site 1 was increased by stimulation of cyclic AMP-dependent protein kinase but not by depolarisation and often occurred in specific nerve terminal sub populations aligned along axon branches. In addition to shedding light on the regulation of synapsin phosphorylation in living nerve terminals, this approach permits the spatially-resolved analysis of the activation of signal transduction pathways in the presynaptic compartment, which is usually too small to be studied with other currently available techniques. PMID- 11017874 TI - Rapid and reciprocal regulation of tenascin-C and tenascin-Y expression by loading of skeletal muscle. AB - Tenascin-C and tenascin-Y are two structurally related extracellular matrix glycoproteins that in many tissues show a complementary expression pattern. Tenascin-C and the fibril-associated minor collagen XII are expressed in tissues bearing high tensile stress and are located in normal skeletal muscle, predominantly at the myotendinous junction that links muscle fibers to tendon. In contrast, tenascin-Y is strongly expressed in the endomysium surrounding single myofibers, and in the perimysial sheath around fiber bundles. We previously showed that tenascin-C and collagen XII expression in primary fibroblasts is regulated by changes in tensile stress. Here we have tested the hypothesis that the expression of tenascin-C, tenascin-Y and collagen XII in skeletal muscle connective tissue is differentially modulated by mechanical stress in vivo. Chicken anterior latissimus dorsi muscle (ALD) was mechanically stressed by applying a load to the left wing. Within 36 hours of loading, expression of tenascin-C protein was ectopically induced in the endomysium along the surface of single muscle fibers throughout the ALD, whereas tenascin-Y protein expression was barely affected. Expression of tenascin-C protein stayed elevated after 7 days of loading whereas tenascin-Y protein was reduced. Northern blot analysis revealed that tenascin-C mRNA was induced in ALD within 4 hours of loading while tenascin-Y mRNA was reduced within the same period. In situ hybridization indicated that tenascin-C mRNA induction after 4 hours of loading was uniform throughout the ALD muscle in endomysial fibroblasts. In contrast, the level of tenascin-Y mRNA expression in endomysium appeared reduced within 4 hours of loading. Tenascin-C mRNA and protein induction after 4-10 hours of loading did not correlate with signs of macrophage infiltration. Tenascin-C protein decreased again with removal of the load and nearly disappeared after 5 days. Furthermore, loading was also found to induce expression of collagen XII mRNA and protein, but to a markedly lower level, with slower kinetics and only partial reversibility. The results suggest that mechanical loading directly and reciprocally controls the expression of extracellular matrix proteins of the tenascin family in skeletal muscle. PMID- 11017875 TI - The cytoplasmic domain of the interleukin-6 receptor gp80 mediates its basolateral sorting in polarized madin-darby canine kidney cells. AB - The IL-6 receptor complex is expressed in different polarized epithelial cells such as liver hepatocytes and intestinal cells. It consists of two subunits: gp80, which binds the ligand, and gp130, which is responsible for signal transduction. In stably transfected Madin-Darby canine kidney (MDCK) cells we have studied the localization of the human IL-6 receptor subunits and found that gp80 and gp130 are predominantly expressed at the basolateral membrane. Analysis of MDCK cells expressing truncated forms of gp80 or gp130 showed that loss of the cytoplasmic domains results in apical delivery. Expression of deletion mutants of gp80 in MDCK cells led to the identification of two discontinous motifs responsible for basolateral sorting: a membrane-proximal tyrosine-based motif (YSLG) and a more membrane-distal dileucine-type motif (LI). Activation of signal transducer and activator of transcription-3 (STAT-3) only occurred via basolaterally located gp80, suggesting that endogenous gp130 is also constrained to the basolateral plasma membrane. Our identification of a basolateral sorting signal within the cytoplasmic region of gp80 for the first time attributes a function to this domain. PMID- 11017876 TI - The apoptosis mediator mDAP-3 is a novel member of a conserved family of mitochondrial proteins. AB - Programmed cell death is essential for organ development and regeneration. To identify molecules relevant for this process, full length cDNA cloning of a short, developmentally regulated murine cDNA fragment, MERM-3, was performed and showed a 1.7 kb mRNA encoding a 45 kDa protein with an ATP/GTP binding motive (P loop). Sequence analysis revealed an 82% amino acid identity to the human death associated protein 3 (hDAP-3), a positive mediator of apoptosis. The full length sequence being the murine orthologue of hDAP-3 is therefore referred to as mDAP 3. In situ hybridization and northern blot analysis showed an abundant mRNA expression with a pronounced expression in highly proliferative epithelial compartments. For mDAP-3, cytochrome c release and induction of cell death could be demonstrated by overexpression of a mDAP-3/EGFP fusion protein. DAP-3 mediated apoptosis was shown to depend on a functional P-loop. Intracellular localization studies using the mDAP-3/EGFP fusion protein, cell fractionation and protease protection experiments localized mDAP-3 to the mitochondrial matrix. DAP-3, in contrast to cytochrome c, retained its mitochondrial localization during apoptosis induction. A mutant of a putative yeast orthologue of mDAP-3, YGL129c, here referred to as yDAP-3, has been shown to exhibit disrupted mitochondrial function. yDAP-3 deficient mutants could be shown to progressively loose mitochondrial DNA. Loss of mitochondrial DNA in yDAP-3 was partially prevented by transfection of the yDAP-3 deficient mutant with mDAP-3, indicating functional complementation by murine DAP-3 in the yeast system. These data identify mDAP-3 as one of the first proapoptotic factors in the mitochondrial matrix and provide evidence for a critical, evolutionary conserved role of members of the DAP-3 protein family for mitochondrial biogenesis. PMID- 11017877 TI - Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. AB - Senescence-associated (beta)-galactosidase is widely used as a biomarker of replicative senescence. However, it remains unknown whether this is a distinct enzyme active at pH 6, and differentially expressed in senescence, or a manifestation of an increase in the classic acid lysosomal (beta)-galactosidase. Here we have investigated the origin of senescence-associated-(beta) galactosidase activity by modifying the intracellular and lysosomal pH of young and senescent human umbilical vein endothelial cells and examining the effect of these manipulations on the levels of activity, using a flow cytometric assay. Lysosomal alkalinisation with chloroquine or bafilomycin A(1), as well as equilibration of the intracellular milieu to pH 6 with nigericin, caused a profound (92-99%) inhibition of the total intracellular (beta)-galactosidase activity. However, independent of pH alterations, senescent cells showed levels of (beta)-galactosidase activity three- to sixfold higher than young cells. This increase in activity occurred in parallel to an increase in (beta)-galactosidase protein levels. Acridine Orange staining revealed an increase in lysosomal content with replicative age, which correlated with the increase in (beta) galactosidase. These findings demonstrate that senescence-associated (beta) galactosidase is a manifestation of residual lysosomal activity at a suboptimal pH, which becomes detectable due to the increased lysosomal content in senescent cells. PMID- 11017878 TI - The p58-positive pre-golgi intermediates consist of distinct subpopulations of particles that show differential binding of COPI and COPII coats and contain vacuolar H(+)-ATPase. AB - We have studied the structural and functional properties of the pre-Golgi intermediate compartment (IC) in normal rat kidney cells using analytical cell fractionation with p58 as the principal marker. The sedimentation profile (sediterm) of p58, obtained by analytical differential centrifugation, revealed in steady-state cells the presence of two main populations of IC elements whose average sedimentation coefficients, s(H)=1150+/-58S ('heavy') and s(L)=158+/-8S ('light'), differed from the s-values obtained for elements of the rough and smooth endoplasmic reticulum. High resolution analysis of these subpopulations in equilibrium density gradients further revealed that the large difference in their s-values was mainly due to particle size. The 'light' particle population contained the bulk of COPI and COPII coats, and redistribution of p58 to these particles was observed in transport-arrested cells, showing that the two types of elements are also compositionally distinct and have functional counterparts in intact cells. Using a specific antibody against the 16 kDa proteolipid subunit of the vacuolar H(+)-ATPase, an enrichment of the V(o )domain of the ATPase was observed in the p58-positive IC elements. Interestingly, these elements could contain both COPI and COPII coats and their density distribution was markedly affected by GTP(&ggr;)S. Together with morphological observations, these results demonstrate that, in addition to clusters of small tubules and vesicles, the IC also consists of large-sized structures and corroborate the proposal that the IC elements contain an active vacuolar H(+)-ATPase. PMID- 11017879 TI - Inhibition of MMH (Met murine hepatocyte) cell differentiation by TGF(beta) is abrogated by pre-treatment with the heritable differentiation effector FGF1. AB - MMH (Met murine hepatocyte) liver cells derived from transgenic mice expressing a truncated constitutively active form of human c-Met are non-transformed immortalized cell lines. We have previously shown that they harbor: (1) epithelial cells that express the liver-enriched transcription factors HNF4 and HNF1(alpha), and that can be stably induced by FGF1 to express liver functions, and (2) fibroblast-like bi-potential palmate cells that can differentiate into bile duct-like structures in Matrigel cultures, or into epithelial cells competent to express hepatic functions. Low concentrations of TGF(beta) have been found to inhibit growth and differentiation of MMH cells. The factor stabilized the palmate cell phenotype, and it provoked epithelial cells to acquire palmate like morphological characteristics, in parallel with down-regulation of expression of HNF4 and HNF1(alpha) and activation of Snail transcripts. The effects of TGF(beta) were dominant if it was added with FGF1, but the effects on differentiation were abrogated if cells had been pre-treated with FGF1. This work identifies TGF(beta) as a factor that could be implicated in maintaining bi potential precursor cells in the liver, FGF1 as one that could over-ride the TGF(beta) effects and Snail as a candidate for mediation of the signal. PMID- 11017880 TI - Regulation of the formation of tumor cell pseudopodia by the Na(+)/H(+) exchanger NHE1. AB - The Na(+)/H(+) exchanger NHE1 is involved in intracellular pH homeostasis and cell volume regulation and accumulates with actin in the lamellipodia of fibroblasts. In order to determine the role of NHE1 following epithelial transformation and the acquisition of motile and invasive properties, we studied NHE1 expression in polarized MDCK cells, Moloney Sarcoma virus (MSV) transformed MDCK cells and an invasive MSV-MDCK cell variant (MSV-MDCK-INV). Expression of NHE1 was significantly increased in MSV-MDCK-INV cells relative to MSV-MDCK and MDCK cells. NHE1 was localized with b-actin to the tips of MSV-MDCK-INV cell pseudopodia by immunofluorescence. Sensitivity of NHE1-mediated (22)Na uptake to ethylisopropylamiloride, a specific inhibitor of NHE1, was increased in MSV-MDCK cells relative to MDCK cells. Changes in intracellular pH induced upon EIPA treatment were also of higher magnitude in MSV-MDCK and MSV-MDCK-INV cells compared to wild-type MDCK cells, especially in Hepes-buffered DMEM medium. Inhibition of NHE1 by 50 microM ethylisopropylamiloride induced the disassembly of actin stress fibers and redistribution of the actin cytoskeleton in all cell types. However, in MSV-MDCK-INV cells, the effect of ethylisopropylamiloride treatment was more pronounced and associated with the increased reversible detachment of the cells from the substrate. Videomicroscopy of MSV-MDCK-INV cells revealed that within 20 minutes of addition, ethylisopropylamiloride induced pseudopodial retraction and inhibited cell motility. The ability of ethylisopropylamiloride to prevent nocodazole-induced formation of actin stress fibers in MSV-MDCK cells was more pronounced in Hepes medium relative to NaHCO(3) medium, showing that NHE1 can regulate actin stress fiber assembly in transformed MSV-MDCK cells via its intracellular pH regulatory effect. These results implicate NHE1 in the regulation of the actin cytoskeleton dynamics necessary for the adhesion and pseudopodial protrusion of motile, invasive tumor cells. PMID- 11017881 TI - Real time imaging reveals a peroxisomal reticulum in living cells. AB - We have directly imaged the dynamic behavior of a variety of morphologically different peroxisomal structures in HepG2 and COS-7 cells transfected with a construct encoding GFP bearing the C-terminal peroxisomal targeting signal 1. Real time imaging revealed that moving peroxisomes interacted with each other and were engaged in transient contacts, and at higher magnification, tubular peroxisomes appeared to form a peroxisomal reticulum. Local remodeling of these structures could be observed involving the formation and detachment of tubular processes that interconnected adjacent organelles. Inhibition of cytoplasmic dynein based motility by overexpression of the dynactin subunit, dynamitin (p50), inhibited the movement of peroxisomes in vivo and interfered with the reestablishment of a uniform distribution of peroxisomes after recovery from nocodazole treatment. Isolated peroxisomes moved in vitro along microtubules in the presence of a microtubule motor fraction. Our data reveal that peroxisomal behavior in vivo is significantly more dynamic and interactive than previously thought and suggest a role for the dynein/dynactin motor in peroxisome motility. PMID- 11017882 TI - Focal adhesion kinase suppresses Rho activity to promote focal adhesion turnover. AB - Focal adhesion kinase (FAK) is activated and localized at focal adhesions upon cell adhesion to extracellular matrices. Cells lacking FAK show increased focal adhesion number and decreased cell migration, functions that are regulated by the small GTPase Rho. We now report that fibroblasts from FAK-/- mice failed to transiently inhibit Rho activity when plated on fibronectin. Re-expression of FAK restored normal Rho regulation. Turnover of focal adhesions correlated inversely with Rho activity. The presence or absence of FAK was mimicked by inhibiting or activating Rho, respectively. These data suggest that loss of FAK resulting in constitutive activation of Rho and inhibition of focal adhesion turnover can account for deficiencies in cell migration and embryonic lethality of the FAK knockout. PMID- 11017883 TI - Effects of 50- or 60-hertz, 100 microT magnetic field exposure in the DMBA mammary cancer model in Sprague-Dawley rats: possible explanations for different results from two laboratories. AB - In line with the possible relationship between electric power and breast cancer risk and the underlying melatonin hypothesis, 50-Hz magnetic field (MF) exposure at microtesla flux densities for either 13 or 27 weeks significantly increased the development and growth of mammary tumors in a series of experiments from Loscher's group in Germany. Loscher's group used the 7,12 dimethylbenz[a]anthracene (DMBA) model of breast cancer in Sprague-Dawley rats. The finding could not be replicated when a similar experimental protocol was used in a study conducted by Battelle in the United States. In the present paper, investigators from the two groups discuss differences between their studies that might explain the apparent discrepancies between the results. These differences include the use of different substrains of Sprague-Dawley rats (the U.S. rats were more susceptible to DMBA than the European rats), different sources for diet and DMBA, differences in environmental conditions, and differences in MF exposure metrics. Furthermore, the effects of MF exposure reported by Loscher's group, albeit significant, were weak. We also discuss the general problem of replicating such weak effects. PMID- 11017884 TI - Exposure to hazardous substances and male reproductive health: a research framework. AB - The discovery in the mid-1970s that occupational exposures to pesticides could diminish or destroy the fertility of workers sparked concern about the effects of hazardous substances on male reproductive health. More recently, there is evidence that sperm quantity and quality may have declined worldwide, that the incidence of testicular cancer has progressively increased in many countries, and that other disorders of the male reproductive tract such as hypospadias and cryptorchidism may have also increased. There is growing concern that occupational factors and environmental chemical exposures, including in utero and childhood exposures to compounds with estrogenic activity, may be correlated with these observed changes in male reproductive health and fertility. We review the evidence and methodologies that have contributed to our current understanding of environmental effects on male reproductive health and fertility and discuss the methodologic issues which confront investigators in this area. One of the greatest challenges confronting researchers in this area is assessing and comparing results from existing studies. We elaborate recommendations for future research. Researchers in the field of male reproductive health should continue working to prioritize hazardous substances; elucidate the magnitude of male reproductive health effects, particularly in the areas of testicular cancer, hypospadias, and cryptorchidism; develop biomarkers of exposure to reproductive toxins and of reproductive health effects for research and clinical use; foster collaborative interdisciplinary research; and recognize the importance of standardized laboratory methods and sample archiving. PMID- 11017885 TI - An assessment of air toxics in Minnesota. AB - We used monitoring and modeling to assess the concentrations of air toxics in the state of Minnesota. Model-predicted concentrations for 148 hazardous air pollutants were from the U.S. Environmental Protection Agency Cumulative Exposure Project (1990 data). Monitoring data consisted of samples of volatile organic compounds, carbonyls, and particulate matter [Less than and equal to] 10 microm in aerodynamic diameter collected at 25 sites throughout the state for varying periods of time (up to 8 years; 1991-1998). Ten pollutants exceeded health benchmark values at one or more sites by modeling, monitoring, or both (including acrolein, arsenic, benzene, 1,3-butadiene, carbon tetrachloride, chromium, chloroform, ethylene dibromide, formaldehyde, and nickel). Polycyclic organic matter also exceeded the benzo[a]pyrene health benchmark value assumed to represent this class of pollutants. The highest modeled and monitored concentrations of most pollutants were near the center of the Minneapolis-St. Paul metropolitan area; however, many smaller cities throughout the state also had elevated concentrations. Where direct comparisons were possible, monitored values often tended to exceed model estimates. Upper-bound excess lifetime inhalation cancer risks were estimated to range from 2.7 [times] 10(-5) to 140. 9 [times] 10(-5) (modeling) and 4.7 [times] 10(-5) to 11.0 [times] 10(-5) (using a smaller set of monitored carcinogens). Screening noncancer hazard indices summed over all end points ranged from 0.2 to 58.1 (modeling) and 0.6 to 2.0 (with a smaller set of monitored pollutants). For common sets of pollutants, the concentrations, cancer risks, and noncancer hazard indices were comparable between model-based estimates and monitored values. The inhalation cancer risk was apportioned to mobile sources (54%), area sources (22%), point sources (12%), and background (12%). This study provides evidence that air toxics are a public health concern in Minnesota. PMID- 11017886 TI - In vivo and in vitro exposure to PCB 153 reduces long-term potentiation. AB - We examined the effects of gestational and lactational exposure to polychlorinated biphenyl (PCB) 153 (2,4,5,2',4',5'-hexaCB) on the magnitude of long-term potentiation (LTP) observed in the CA1 region of hippocampal brain slices prepared from rats at 30 days of age. We compared these actions to those observed when PCB 153 is dissolved in normal Krebs-Ringer solution and perfused on slices from control rats of the same age. In vivo exposure was at three dose levels (1. 25, 5, and 20 mg/kg/day) from gestational day 3 through weaning at postnatal day 21. Although responses to low-frequency stimulation of the Schaffer collateral pathway in exposed animals were not different from controls, significantly reduced LTP was induced after tetanic stimulation, even at the lowest dose studied. We observed a comparable depression of LTP when control slices were perfused with Krebs-Ringer that had been equilibrated with PCB 153 in a generator column. Neither in vivo nor in vitro exposure significantly altered the input-output curves obtained before tetanic stimulation, but both suppressed the increase in response observed in controls after tetanic stimulation. Because LTP is thought to be correlated with learning ability, these observations may provide at least a partial mechanism to explain the reduction of intelligence quotient observed in humans exposed to PCBs early in development. PMID- 11017887 TI - The contributions of emissions and spatial microenvironments to exposure to indoor air pollution from biomass combustion in Kenya. AB - Acute and chronic respiratory diseases, which are causally linked to exposure to indoor air pollution in developing countries, are the leading cause of global morbidity and mortality. Efforts to develop effective intervention strategies and detailed quantification of the exposure-response relationship for indoor particulate matter require accurate estimates of exposure. We used continuous monitoring of indoor air pollution and individual time-activity budget data to construct detailed profiles of exposure for 345 individuals in 55 households in rural Kenya. Data for analysis were from two hundred ten 14-hour days of continuous real-time monitoring of concentrations of particulate matter [less than/equal to] 10 microm in aerodynamic diameter and the location and activities of household members. These data were supplemented by data on the spatial dispersion of pollution and from interviews. Young and adult women had not only the highest absolute exposure to particulate matter (2, 795 and 4,898 microg/m(3) average daily exposure concentrations, respectively) but also the largest exposure relative to that of males in the same age group (2.5 and 4.8 times, respectively). Exposure during brief high-intensity emission episodes accounts for 31-61% of the total exposure of household members who take part in cooking and 0-11% for those who do not. Simple models that neglect the spatial distribution of pollution within the home, intense emission episodes, and activity patterns underestimate exposure by 3-71% for different demographic subgroups, resulting in inaccurate and biased estimations. Health and intervention impact studies should therefore consider in detail the critical role of exposure patterns, including the short periods of intense emission, to avoid spurious assessments of risks and benefits. PMID- 11017888 TI - Are there sensitive subgroups for the effects of airborne particles? AB - Recent studies have shown that particulate air pollution is a risk factor for hospitalization for heart and lung disease; however, little is known about what subpopulations are most sensitive to this pollutant. We analyzed Medicare hospital admissions for heart disease, chronic obstructive pulmonary disorders (COPD) and pneumonia in Chicago, Cook County, Illinois, between 1985 and 1994. We examined whether previous admissions or secondary diagnoses for selected conditions predisposed persons to having a greater risk from air pollution. We also considered effect modification by age, sex, and race. We found that the air pollution-associated increase in hospital admissions for cardiovascular diseases was almost doubled in subjects with concurrent respiratory infections. The risk was also increased by a previous admission for conduction disorders. For COPD and pneumonia admissions, diagnosis of conduction disorders or dysrhythmias increased the risk of particulate matter < 10 microm in aerodynamic diameter (PM(10)) associated admissions. Persons with asthma had twice the risk of a PM(10) associated pneumonia admission and persons with heart failure had twice the risk of PM(10)-induced COPD admissions. The PM(10) effect did not vary by sex, age, and race. These results suggest that patients with acute respiratory infections or defects in the electrical control of the heart are a risk group for particulate matter effects. PMID- 11017889 TI - Long-term arsenic exposure and incidence of non-insulin-dependent diabetes mellitus: a cohort study in arseniasis-hyperendemic villages in Taiwan. AB - Diabetes prevalence in arseniasis-hyperendemic villages in Taiwan has been reported to be significantly higher than in the general population. The aim of this cohort study was to further evaluate the association between ingested inorganic arsenic and the incidence of non-insulin-dependent diabetes mellitus in these villages. A total of 446 nondiabetic residents in these villages were followed biannually by oral glucose tolerance test. Diabetes is defined as a fasting plasma glucose level > or = 7.8 mmol/L and/or a 2-hr post-load glucose level > or = 11.1 mmol/L. During the follow-up period of 1499.5 person-years, 41 cases developed diabetes, showing an overall incidence of 27.4/1,000 person years. The incidence of diabetes correlated with age, body mass index, and cumulative arsenic exposure. The multivariate-adjusted relative risks were 1.6, 2.3, and 2.1 for age > or = 55 versus < 55 years, a body mass index ?Greater/Equal to] 25 versus < 25 kg/m(2), and a cumulative arsenic exposure > or = 17 versus < 17 mg/L-years, respectively. The incidence density ratios (95% confidence intervals) between the hyperendemic villages and the two nonendemic control townships were 3.6 (3.5-3.6), 2.3 (1.1-4.9), 4.3 (2.4-7.7), and 5.5 (2.2 13.5), respectively, for the age groups of 35-44, 45-54, 55-64, and 65-74 years. The findings are consistent with our previous cross-sectional observation that ingested inorganic arsenic is diabetogenic in human beings. PMID- 11017890 TI - Real-time and integrated measurement of potential human exposure to particle bound polycyclic aromatic hydrocarbons (PAHs) from aircraft exhaust. AB - We used real-time monitors and low-volume air samplers to measure the potential human exposure to airborne polycyclic aromatic hydrocarbon (PAH) concentrations during various flight-related and ground-support activities of C-130H aircraft at an Air National Guard base. We used three types of photoelectric aerosol sensors (PASs) to measure real-time concentrations of particle-bound PAHs in a break room, downwind from a C-130H aircraft during a four-engine run-up test, in a maintenance hangar, in a C-130H aircraft cargo bay during cargo-drop training, downwind from aerospace ground equipment (AGE), and in a C-130H aircraft cargo bay during engine running on/off (ERO) loading and backup exercises. Two low volume air samplers were collocated with the real-time monitors for all monitoring events except those in the break room and during in-flight activities. Total PAH concentrations in the integrated-air samples followed a general trend: downwind from two AGE units > ERO-loading exercise > four-engine run-up test > maintenance hangar during taxi and takeoff > background measurements in maintenance hangar. Each PAH profile was dominated by naphthalene, the alkyl substituted naphthalenes, and other PAHs expected to be in the vapor phase. We also found particle-bound PAHs, such as fluoranthene, pyrene, and benzo[a]pyrene in some of the sample extracts. During flight-related exercises, total PAH concentrations in the integrated-air samples were 10-25 times higher than those commonly found in ambient air. Real-time monitor mean responses generally followed the integrated-air sample trends. These monitors provided a semiquantitative temporal profile of ambient PAH concentrations and showed that PAH concentrations can fluctuate rapidly from a baseline level < 20 to > 4,000 ng/m(3) during flight-related activities. Small handheld models of the PAS monitors exhibited potential for assessing incidental personal exposure to particle-bound PAHs in engine exhaust and for serving as a real-time dosimeter to indicate when respiratory protection is advisable. PMID- 11017891 TI - Survey of bottled drinking water available in Manitoba, Canada. AB - Forty domestic and imported brands of bottled water were purchased in Manitoba, Canada and examined for total dissolved solids (TDS), chloride, sulfate, nitrate nitrogen, cadmium, lead, copper, and radioactivity. The samples showed great variation in quality, and some exceeded the Canadian Water Quality Guidelines for drinking water for TDS, chloride, and lead. Carbonation, ozonation, and type of packaging were not associated with differences in metal levels, although carbonated samples tended to show higher TDS values. A number of deficiencies were found with respect to product labeling. PMID- 11017892 TI - Interactions of dietary estrogens with human estrogen receptors and the effect on estrogen receptor-estrogen response element complex formation. AB - Epidemiologic and experimental studies support the hypothesis that dietary estrogens from plant sources (phytoestrogens) may play a role in the prevention of breast and prostate cancer. The molecular mechanisms for such chemopreventive effect are still unclear. We investigated the possibility that phytoestrogens may bind differentially to estrogen receptor proteins (ER[alpha] and ERss) and affect the interactions of the ligand-ER complexes with different estrogen response element (ERE) sequences. We used fluorescence polarization to measure the binding affinities of genistein, coumestrol, daidzein, glyceollin, and zearalenone for human ER[alpha] and ERss. Competition binding experiments revealed higher affinity of the phytoestrogens for ERss than for ER[alpha]. Genistein [median inhibitory concentration 12nM] is the most potent and has the same relative binding affinity for ERss as 17ss-estradiol. We also studied the effect of these phytoestrogens on the ability of ER[alpha] and ERss to associate with specific DNA sequences (EREs). The direct binding of human recombinant estrogen receptors to fluorescein-labeled EREs indicates that phytoestrogens can cause conformational changes in both human ERs, which results in altered affinities of the complexes for the ERE from the Xenopus vitellogenin A2 gene and an ERE from the human pS2 gene. PMID- 11017893 TI - Cancer mortality in four northern wheat-producing states. AB - Chlorophenoxy herbicides are used both in cereal grain agriculture and in nonagricultural settings such as right-of-ways, lawns, and parks. Minnesota, North Dakota, South Dakota, and Montana grow most of the spring and durum wheat produced in the United States. More than 90% of spring and durum wheat is treated with chlorophenoxy herbicides, in contrast to treatment of approximately 30% of winter wheat. In this ecologic study I used wheat acreage as a surrogate for exposure to chlorophenoxy herbicides. I investigated the association of chlorophenoxy herbicides with cancer mortality during 1980-1989 for selected counties based on level of agriculture ([greater and equal to] 20%) and rural population ([greater and equal to] 50%). Age-standardized cancer mortality rates were determined for grouped counties based on tertiles of wheat acreage per county or for individual counties for frequently occurring cancers. The cancer sites that showed positive trends of increasing cancer mortality with increasing wheat acreage were esophagus, stomach, rectum, pancreas, larynx, prostate, kidney and ureter, brain, thyroid, bone, and all cancers (men) and oral cavity and tongue, esophagus, stomach, liver and gall bladder and bile ducts, pancreas, cervix, ovary, bladder, and other urinary organs, and all cancers (women). Rare cancers in men and women and cancers in boys and girls were studied by comparing counties above and below the median of wheat acreage per county. There was increased mortality for cancer of the nose and eye in both men and women, brain and leukemia in both boys and girls, and all cancers in boys. These results suggest an association between cancer mortality and wheat acreage in counties of these four states. PMID- 11017894 TI - Relation between stillbirth and specific chlorination by-products in public water supplies. AB - During water treatment, chlorine reacts with naturally occurring organic matter in surface water to produce a number of by-products. Of the by-products formed, trihalomethanes (THMs) are among the highest in concentration. We conducted a retrospective cohort study to evaluate the relationship between the level of total THM and specific THMs in public water supplies and risk for stillbirth. The cohort was assembled from a population-based perinatal database in the Canadian province of Nova Scotia and consisted of almost 50,000 singleton deliveries between 1988 and 1995. Individual exposures were assigned by linking mother's residence at the time of delivery to the levels of specific THMs monitored in public water supplies. Analysis was conducted for all stillbirths and for cause of-death categories based on the physiologic process responsible for the fetal death. Total THMs and the specific THMs were each associated with increased stillbirth risk. The strongest association was observed for bromodichloromethane exposure, where risk doubled for those exposed to a level of [greater and equal to] 20 microg/L compared to those exposed to a level < 5 microg/L (relative risk = 1. 98, 95% confidence interval, 1.23-3.49). Relative risk estimates associated with THM exposures were larger for asphyxia-related deaths than for unexplained deaths or for stillbirths overall. These findings suggest a need to consider specific chlorination by-products in relation to stillbirth risk, in particular bromodichloromethane and other by-product correlates. The finding of a stronger effect for asphyxia deaths requires confirmation and research into possible mechanisms. PMID- 11017895 TI - Semen quality and reproductive health of young Czech men exposed to seasonal air pollution. AB - This study of male reproductive health in the Czech Republic resulted from community concern about potential adverse effects of air pollution. We compared young men (18 years of age) living in Teplice, a highly industrialized district with seasonally elevated levels of air pollution, to those from Prachatice, a rural district with relatively clean air. Surveys were scheduled for either late winter, after the season of higher air pollution, or at the end of summer, when pollution was low. Participation included a physical examination, donation of a semen sample, and completion of a questionnaire on health, personal habits, and exposure to solvents and metals through work or hobby. Analysis of data from 408 volunteers showed that the men from Teplice and Prachatice were similar in physical characteristics, personal habits, and work- or hobby-related exposures. Sixty-six percent (272) of these men donated a single semen sample for routine semen analysis, computer-aided sperm motion analysis, and sperm chromatin structure assay. The mean (median) sperm concentration and sperm count were 61. 2 (44.0) million/mL semen and 113.3 (81.5) million, respectively, and were not associated with district of residence or period of elevated air pollution. However, periods of elevated air pollution in Teplice were significantly associated with decrements in other semen measures including proportionately fewer motile sperm, proportionately fewer sperm with normal morphology or normal head shape, and proportionately more sperm with abnormal chromatin. These results suggest that young men may experience alterations in sperm quality after exposure to periods of elevated air pollution, without changes in sperm numbers. PMID- 11017896 TI - Identification of phthalate esters in the serum of young Puerto Rican girls with premature breast development. AB - Premature breast development (thelarche) is the growth of mammary tissue in girls younger than 8 years of age without other manifestations of puberty. Puerto Rico has the highest known incidence of premature thelarche ever reported. In the last two decades since this serious public health anomaly has been observed, no explanation for this phenomenon has been found. Some organic pollutants, including pesticides and some plasticizers, can disrupt normal sexual development in wildlife, and many of these have been widely used in Puerto Rico. This investigation was designed to identify pollutants in the serum of Puerto Rican girls with premature thelarche. A method for blood serum analysis was optimized and validated using pesticides and phthalate esters as model compounds of endocrine-disrupting chemicals. Recovery was > 80% for all compounds. We performed final detection by gas chromatography/mass spectrometry. We analyzed 41 serum samples from thelarche patients and 35 control samples. No pesticides or their metabolite residues were detected in the serum of the study or control subjects. Significantly high levels of phthalates [dimethyl, diethyl, dibutyl, and di-(2-ethylhexyl)] and its major metabolite mono-(2-ethylhexyl) phthalate were identified in 28 (68%) samples from thelarche patients. Of the control samples analyzed, only one showed significant levels of di-isooctyl phthalate. The phthalates that we identified have been classified as endocrine disruptors. This study suggests a possible association between plasticizers with known estrogenic and antiandrogenic activity and the cause of premature breast development in a human female population. PMID- 11017897 TI - Susceptibility in microbial risk assessment: definitions and research needs. PMID- 11017898 TI - Applying biomarker research. PMID- 11017899 TI - Modeling of exposure to carpet-cleaning chemicals preceding irritant-induced asthma in one patient. AB - 42-year-old woman experienced an acute asthma attack, seizures, and unconsciousness immediately after a carpet-cleaning and deodorizing job was conducted in her home. Exposure modeling estimates that she was exposed to approximately 3.4-17 mg/m(3) of sodium tripolyphosphate and more than 14 mg/m(3) volatile organic compounds immediately after the cleaning. I derived two separate exposure models for these estimates that evidenced good consistency of exposure estimates. Asthmatics and carpet-cleaning companies should be advised about safety during carpet-cleaning operations, including adequate warnings about excess risk for asthmatics, temporary removal from the home, reduced detergent levels within cleaners, and reduced overall levels of cleaning solutions used within the home. Further studies of carpet-cleaning exposures are indicated. PMID- 11017900 TI - Evaluating scientific impact. PMID- 11017901 TI - Reflections on hexavalent chromium: health hazards of an industrial heavyweight. AB - Chromium has been used commercially in the United States for more than 100 years in metal alloys and other compounds, as a pigment, and in the tanning and metal plating industries, and many studies have looked at its effects in terms of occupational health. But, although scientists know that Cr(VI) is a human carcinogen and that it can cause other deleterious health effects including kidney and liver damage, certain questions remain about the metal's effects, such as which routes of exposures are dangerous for humans. PMID- 11017902 TI - A new crop of concerns: Congress investigates pesticide safety. AB - A March 2000 report by the General Accounting Office, Pesticides: Improvements Needed to Ensure the Safety of Farmworkers and Their Children, states that much remains unknown about the risks faced by children in agriculture, and that enforcement of pesticide protection standards for farmworkers is patchy and unsystematic. Many cases of farmworkers' pesticide-related illnesses go unreported, leaving health workers with an inadequate basis for tracking patterns and fine-tuning pesticide standards, says the report. In addition, children are known to be more vulnerable to the effects of pesticides, but there is a lack of data regarding children's exposures and the precise effects of pesticides on children's health. PMID- 11017903 TI - Go with the flow: an updated tool for detecting molecules. AB - In the early 1970, researchers at Los Alamos National Laboratories developed the flow cytometer, a device that allows for the identification of unknown cells. In a flow cytometer, a single-cell suspension is passed in a continuous flow through a laser beam, with each cell scattering the light in a characteristic manner. A few years ago, researchers at Los Alamos began another project, refining the capabilities of the flow cytometer so that it could analyze not a single cell but a single molecule, allowing scientists to study bacteria at the molecular level, differentiating between individual strains more quickly and with greater accuracy than before. The new flow cytometer shows great promise in a variety of fields where single-molecule study would be valuable, such as genomics and disease transmission. PMID- 11017904 TI - Phosphatidylinositol kinase regulation of airway smooth muscle cell proliferation. PMID- 11017905 TI - Hyaluronan: is bigger better? PMID- 11017906 TI - Role of alveolar macrophages in host defense against Pneumocystis carinii. PMID- 11017907 TI - Regulation of cyclin D(1) expression and DNA synthesis by phosphatidylinositol 3 kinase in airway smooth muscle cells. AB - We have shown in bovine tracheal myocytes that extracellular signal-regulated kinase (ERK) and Rac1 function as upstream activators of transcription from the cyclin D(1) promoter. We now examine the role of phosphatidylinositol (PI) 3 kinase in this process. PI 3-kinase activity was increased by platelet-derived growth factor (PDGF) and attenuated by the PI 3-kinase inhibitors wortmannin and LY294002. These inhibitors also decreased cyclin D(1) promoter activity, protein abundance, and DNA synthesis. Overexpression of the active catalytic subunit of PI 3-kinase (p110(PI) (3-K)CAAX) was sufficient to activate the cyclin D(1) promoter. Wortmannin and LY294002 failed to attenuate PDGF-induced ERK activation, and overexpression of p110(PI) (3-K)CAAX was insufficient to activate ERK. p110(PI) (3-K)CAAX-induced cyclin D(1) promoter activity was not blocked by PD98059, an inhibitor of mitogen-activated protein kinase/ERK kinase. We next examined whether PI 3-kinase and the 21-kD guanidine triphosphatase Rac1 regulate cyclin D(1) promoter activity by similar mechanisms. p110(PI) (3-K)CAAX-induced cyclin D(1) promoter activity was decreased by two inhibitors of Rac1-mediated signaling, catalase and diphenylene iodonium. Further, PDGF, PI 3-kinase, and Rac1 each activated the cyclin D(1) promoter at the cyclic adenosine monophosphate response element binding protein (CREB)/activating transcription factor (ATF)-2 binding site, as evidenced by expression of a CREB/ATF-2 reporter plasmid. Finally, PI 3-kinase and Rac1-induced CREB/ATF-2 transactivation were each inhibited by catalase. Together, these data suggest that in airway smooth muscle (ASM) cells, PI 3-kinase regulates transcription from the cyclin D(1) promoter and DNA synthesis in an ERK-independent manner. Further, PI 3-kinase and Rac1 regulate ASM cell cycle traversal via a common cis-regulatory element in the cyclin D(1) promoter. PMID- 11017908 TI - Activation and transforming growth factor-beta production in eosinophils by hyaluronan. AB - To investigate whether extracellular matrix glycosaminoglycan hyaluronan (HA) modulates eosinophil activation and transforming growth factor (TGF)-beta production by eosinophils, human peripheral blood eosinophils (purity > 99%) from 12 patients with mild to moderate asthma or six healthy subjects were isolated and incubated with increasing concentrations of low molecular weight (mol wt) HA ( approximately 0.2 x 10(6) D) or high mol wt HA (3.0 to approximately 5.8 x 10(6) D). We found that the low mol wt HA has a pronounced effect on eosinophil survival in both patients with asthma and healthy subjects in a dose-dependent fashion on Days 2 and 4. Whereas the high mol wt HA had a smaller effect on eosinophil survival than did the low mol wt HA. The HA-mediated eosinophil survival was partially but significantly inhibited ( approximately 50% inhibition) by a blocking monoclonal antibody for CD44, a specific receptor of HA, and largely inhibited by an anti-granulocyte macrophage colony-stimulating factor (GM-CSF) neutralizing antibody but not by an anti-interleukin (IL)-3 or anti-IL-5 neutralizing antibody. In addition, the low mol wt HA increased GM-CSF messenger RNA (mRNA) expression and protein secretion by eosinophils in a dose dependent fashion, suggesting that the HA-mediated eosinophil survival is due mainly to induction of GM-CSF release through partial CD44 signaling. Furthermore, we demonstrated that the low mol wt HA results in morphologic changes in eosinophils such as transforming from a round to a spindle shape and in homotypic aggregation, upregulates intercellular adhesion molecule-1 expression, and increases TGF-beta mRNA expression and protein secretion by eosinophils. These observations suggest previously unforeseen interactions between eosinophils and low mol wt extracellular matrix and, thus, novel pathways by which eosinophils may contribute to the regulation of airway inflammation and airway remodeling. PMID- 11017909 TI - Alveolar macrophages from human immunodeficiency virus-infected persons demonstrate impaired oxidative burst response to Pneumocystis carinii in vitro. AB - The alveolar macrophage (AM) oxidative burst response is an important component of microbicidal effector cell function against a variety of potential pathogens in the lungs, although the role against Pneumocystis carinii has not been fully investigated. The goals of this study were to characterize the P. carinii mediated oxidative burst of AMs from healthy individuals, and to examine the oxidative burst of AMs from human immunodeficiency virus (HIV)-infected persons. For healthy individuals, the AM oxidative burst (measured as hydrogen peroxide [H(2)O(2)] production) increased in a time- and concentration-dependent manner in response to P. carinii or to the major surface glycoprotein of P. carinii, gp-A (0.01 to 10 microg/ml), required physical contact of P. carinii with AMs, and was not dependent on organism viability. Enzymatic removal of the surface-associated molecules of P. carinii reduced the oxidative burst to 43% of control (P = 0.01). Blocking the AM mannose receptor reduced the P. carinii-mediated oxidative burst response to 37% of control (P = 0.01). Compared with AMs from healthy individuals, P. carinii-mediated H(2)O(2) production was significantly reduced in AMs from asymptomatic HIV-positive (HIV+) persons with CD4+ counts < 200 cells/mm(3) (249+/-43 relative fluorescence units [RFU] versus 130+/-44 RFU; mean +/- standard error of the mean, P = 0.038) and HIV+ persons with active P. carinii pneumonia (78+/-40 RFU; P = 0.014), but preserved for HIV+ persons with CD4+ counts > 200 cells/mm(3). Importantly, H2O2 production in response to phorbol myristate acetate or serum-opsonized zymosan particles was preserved in all groups studied. Thus, AM oxidative burst, mediated in part via P. carinii gp A and AM mannose receptor may represent an important host response to P. carinii. A specific impairment of P. carinii-mediated AM oxidative burst in persons with advanced HIV infection may contribute to the pathogenesis of P. carinii pneumonia. PMID- 11017910 TI - Induction of telomerase activity in fibroblasts from bleomycin-injured lungs. AB - Bleomycin-induced lung injury causes increased fibroblast numbers in the lung and pulmonary fibrosis. Studies of fibroblasts isolated from such injured lungs have revealed evidence of increased intrinsic proliferative capacity, but the mechanism is unknown. Telomerase catalyzes the addition of telomeric DNA repeats onto chromosomal ends, which is associated with increased cellular life span or immortality. To examine whether telomerase might play a role in regulating fibroblast proliferative capacity in pulmonary fibrosis, lung fibroblasts were isolated from rats treated with endotracheal injections of phosphate-buffered saline or bleomycin. At selected time points, the rats were killed and lung fibroblasts isolated. The isolated cells and lung tissue were then used in experiments for measurement of telomerase activity. The results show undetectable telomerase activity in fibroblasts isolated from control uninjured lungs, or in the control lung tissue extracts. Similar results were obtained in cells and lung tissue from Days 1, 3, and 28 bleomycin-injured lungs. However, significant telomerase activity was detected in fibroblasts and tissue extracts isolated from Days 7, 14, and 21 bleomycin-treated rat lungs, with maximal activity observed in the Day 14 samples. Analysis of the isolated cells for telomerase messenger RNA or reverse transcriptase expression, combined with alpha-smooth-muscle actin expression by immunohistochemistry, revealed that telomerase expression localized primarily to nonmyofibroblasts. These findings suggest that in addition to elevated growth factor expression, the injured lung fibroblast population may contain cells with increased life span, which could contribute to the observed overall increase in lung fibroblast numbers. PMID- 11017911 TI - Differential gene expression in the initiation and progression of nickel-induced acute lung injury. AB - Acute lung injury, an often fatal condition, can result from a wide range of insults leading to a complex series of biologic responses. Despite extensive research, questions remain about the interplay of the factors involved and their role in acute lung injury. We proposed that assessing the temporal and functional relationships of differentially expressed genes after pulmonary insult would reveal novel interactions in the progression of acute lung injury. Specifically, 8,734 sequence-verified murine complementary DNAs were analyzed in mice throughout the initiation and progression of acute lung injury induced by particulate nickel sulfate. This study revealed the expression patterns of genes previously associated with acute lung injury in relationship to one another and also uncovered changes in expression of a number of genes not previously associated with acute lung injury. The overall pattern of gene expression was consistent with oxidative stress, hypoxia, cell proliferation, and extracellular matrix repair, followed by a marked decrease in pulmonary surfactant proteins. Also, expressed sequence tags (ESTs), with nominal homology to known genes, displayed similar expression patterns to those of known genes, suggesting possible roles for these ESTs in the pulmonary response to injury. Thus, this analysis of the progression and response to acute lung injury revealed novel gene expression patterns. PMID- 11017912 TI - A role for hyaluronan in macrophage accumulation and collagen deposition after bleomycin-induced lung injury. AB - Elevated concentrations of hyaluronan (HA) are associated with the accumulation of macrophages in the lung after injury. We have investigated the role of HA in the inflammatory and fibrotic responses to lung injury using the intratracheal instillation of bleomycin in rats as a model. After bleomycin-induced lung injury, both HA content in bronchoalveolar lavage (BAL) and staining for HA in macrophages accumulating in injured areas of the lung were maximal at 4 d. Increased HA in BAL correlated with increased locomotion of isolated alveolar macrophages. HA-binding peptide was able to specifically block macrophage motility in vitro. Importantly, systemic administration of HA-binding peptide to rats before injury not only decreased alveolar macrophage motility and accumulation in the lung, but also reduced lung collagen alpha (I) messenger RNA and hydroxyproline contents. We propose a model in which HA plays a critical role in the inflammatory response and fibrotic consequences of acute lung injury. PMID- 11017913 TI - Induction of apoptosis by glyoxal in human embryonic lung epithelial cell line L132. AB - Oxidative stress has been suggested to play a central role in the pathogenesis of lung fibrosis and lung epithelial cell apoptosis is considered to be a key event during fibrogenesis. Studies from various laboratories have indicated that metabolic conditions may initiate oxidative stress, thereby contributing to epithelial cell death. This study was designed to test the hypothesis that glyoxal, an intermediate product in the glycation reaction leading to advanced glycation end products (AGEs), may induce lung epithelial cell apoptosis. We investigated the in vitro effects of glyoxal on fetal human lung epithelial L132 cells. Immunocytochemical analysis of paraffin-embedded cells and fluorescence activated cell sorter analysis revealed a dose-dependent accumulation of the glycoxidation product (epsilon)N-carboxymethyllysine (CML) in all compartments of the cell. It has been shown that CML modification of proteins may serve as an indicator for oxidative stress. To examine the role of apoptosis in epithelial lung cells we investigated glyoxal-dependent changes in pro- and antiapoptotic mediators bax and activated caspase-3, and galectin-3 and bcl-2, respectively. Increasing concentrations of glyoxal (50 to 400 microM) induced an increase in the number of apoptotic cells. The apoptotic changes were confirmed by transmission electron microscopy. Immunocytochemical analysis of treated cells revealed the presence of other AGEs such as pentosidine as well as products of lipid peroxidation. PMID- 11017914 TI - Endothelial sulfated sialyl Lewis x glycans, putative L-selectin ligands, are preferentially expressed in bronchial asthma but not in other chronic inflammatory lung diseases. AB - Lymphocyte infiltrate is a hallmark of inflammatory responses. We have previously shown that de novo-induced endothelial sialyl Lewis x (sLex) expression guides lymphocytes in an L-selectin-dependent manner to sites of acute organ transplant rejections. In this research, we have analyzed five groups of chronic lung inflammations to determine the presence of properly glycosylated, i.e., sulfated, sLex-decorated, L-selectin ligands. Two anti-sLex (2F3 and HECA-452) and one anti 6- and/or 6'-sulfated and/or 6,6'-bisulfated (MECA-79) monoclonal antibodies (mAbs) were used. The control lung specimens did not express L-selectin ligands on endothelium. In contrast, the endothelial staining intensity and the number of positive peribronchial venules and capillaries with mAbs 2F3, HECA-452, and MECA 79 were significantly greater in bronchial biopsies from patients with asthma compared with normal specimens (P<0.003). However, no significant increase of peribronchial endothelial reactivity with these antibodies was observed in adult respiratory distress syndrome, chronic bronchitis, fibrosing alveolitis, and granulomatous inflammation compared with controls. These data suggest that sulfated sLex glycans, acting putatively as ligands for L-selectin, could be instrumental in lymphocyte extravasation into human peribronchial lung tissue during asthma, but not so important in several other inflammatory lung diseases. PMID- 11017915 TI - Instillation of allogeneic lung antigen-presenting cells deficient in expression of major histocompatibility complex class I or II antigens have differential effects on local cellular and humoral immunity and on pathology in recipient murine lungs. AB - Recognition of allogeneic major histocompatibility complex (MHC) molecules expressed on donor lung antigen-presenting cells (APCs) by host T lymphocytes is believed to stimulate lung allograft rejection. However, the specific roles of donor MHC molecules in the rejection response is unknown. We report a murine model in which instilling allogeneic lung APCs into recipient lungs induces pathology analogous to acute rejection, and the production of interferon (IFN) gamma, immunoglobulin (Ig) G2a, and alloantibodies in recipient lungs. Using allogeneic lung APCs (C57BL/6, I-a(b), H-2(b)) deficient in MHC class I, II, or both for instillation into lungs of BALB/c mice (I-a(d), H-2(d)), the purpose of the current study was to determine the specific roles of donor MHC molecules in stimulating local alloimmune responses. The data show that MHC class I or II on donor APCs induced IFN-gamma and IgG2a synthesis locally, though less than that induced by wild-type cells. Both MHC class I and II were required to induce alloantibody production. Instillation of wild-type or class I- or class II deficient APCs induced comparable pathologic lesions in recipient lungs, and more severe than that induced by MHC-deficient cells. These data show that donor MHC class I and II molecules have differential effects in the stimulation of local alloimmune responses. PMID- 11017916 TI - DNA from bronchial secretions modulates elastase inhibition by alpha(1) proteinase inhibitor and oxidized secretory leukoprotease inhibitor. AB - Previously we reported that DNA from sputum promotes the inhibition of human leukocyte elastase (HLE) by native secretory leukoprotease inhibitor (SLPI). This study shows that sputum DNA also promotes the inhibition by oxidized SLPI, a form of SLPI that may occupy a large fraction of the inhibitor in the lungs under conditions of high oxidative stress. With sputum DNA at 5 microg/ml, a concentration much lower than those in vivo, the inhibition constant (K(i) ) of oxidized SLPI against HLE is reduced from 31 nM to 23 to 920 pM, as compared with the K(i) of native SLPI, 58 pM, under the same conditions. On the other hand, sputum DNA retards inhibition of HLE by alpha(1)-proteinase inhibitor (alpha(1) PI). The association rate of alpha(1)-PI and HLE is decreased from 1 x 10(7) M( 1) s(-1) in the absence of DNA to 2 to 6 x 10(6) M(-1) s(-1) in the presence of sputum DNA at 100 microg/ml. On the basis of results with an elastase-specific oligonucleotide aptamer, it was found that the downregulation of alpha(1)-PI activity can be attributed to an interaction between sputum DNA and multiple DNA binding sites on HLE. DNA-binding sites on HLE also participate in the upregulation of oxidized SLPI activity. Data from this and our previous studies demonstrate that sputum DNA facilitates the association of HLE with native and oxidized SLPI, whereas it delays the association of HLE with alpha(1)-PI. We conclude that by modulating the inhibition of HLE, sputum DNA directly affects the balance between proteases and antiproteases in the lungs. PMID- 11017917 TI - Inositol (1,4,5)trisphosphate metabolism and enhanced calcium mobilization in airway smooth muscle of hyperresponsive rats. AB - Airway hyperresponsiveness (AHR) is a phenotype of asthma and can be modeled by the inbred Fisher strain of rat, which is hyperresponsive in vivo relative to the Lewis strain. Enhanced airway smooth muscle (ASM) contractility and Ca(2+) mobilization are associated with the AHR observed in Fisher rats. In this study, we investigated whether the interstrain differences in Ca(2+) mobilization to serotonin (5HT) result from differences in inositol (1,4,5)trisphosphate (IP(3)) metabolism and/or IP(3) receptor (IP(3)R) sensitivity. Ca(2+) mobilization by 5HT in cultured ASM cells from both rat strains was phospholipase C (PLC) dependent. Inositol polyphosphate accumulation, and hence PLC activity, was similar in both rat strains, but a specific IP(3) transient was detectable only in Fisher myocytes in response to 5HT. These findings suggested that IP(3) degradation rather than production differed between the two strains. The Vmax and Michaelis constant (K(m)) of IP(3)-specific 5-phosphatase activity were higher in the particulate fraction of Lewis than in Fisher ASM cell homogenates and appeared to be related to a greater expression of two isoforms of 5-phosphatase (type I and type II) in Lewis cells as shown by Western blot analysis. The sensitivity of the IP(3)R to IP(3) was similar between Fisher and Lewis ASM cells, indicating that the interstrain intracellular Ca(2+) differences were unrelated to IP(3)R function. We propose that interstrain variations in 5-phosphatase activity and expression may give rise to the interstrain differences in IP(3)-mediated Ca(2+) release in ASM and may be a determinant of AHR. PMID- 11017918 TI - Piecemeal degranulation of peripheral blood eosinophils: a study of allergic subjects during and out of the pollen season. AB - The variability of serum and plasma levels of eosinophil granule proteins in different clinical conditions, interpreted as the result of different patterns of cytokine priming, suggests a selective mobilization of granule proteins. Inasmuch as piecemeal degranulation (PM) is the mechanism proposed for the differential release of eosinophil granule proteins, we decided to investigate whether blood eosinophils from allergic subjects show characteristics of PM during natural allergen challenge. Eosinophils from three birch-sensitive subjects were studied before and during the pollen season. Electron microscopy analysis showed that during the season, eosinophils presented morphologic features of PM. By immunogold labeling, eosinophil cationic protein (ECP) was detected not only in normal specific granules but also in the cytoplasm, in the vicinity of partially lucent specific granules. These results were confirmed by subcellular fractionation, where the amount of ECP associated with compartments containing small vesicles increased 2-fold during the pollen season. A study of the distribution of ECP, eosinophil peroxidase, and hexosaminidase in eosinophils of different densities showed that the profile of each of these proteins differed depending on cell density. All of these proteins decreased in the specific granule of hypodense cells and increased in other cell compartments. We conclude that allergen exposure causes PM of the peripheral blood eosinophils of allergic subjects, and that the density of these cells reflects the degree of degranulation. Our results provide novel information for the understanding of the selective mobilization of granule proteins into the circulation. PMID- 11017919 TI - Effect of cigarette smoke on the permeability and IL-1beta and sICAM-1 release from cultured human bronchial epithelial cells of never-smokers, smokers, and patients with chronic obstructive pulmonary disease. AB - Although cigarette smoking is of paramount importance in the development of chronic obstructive pulmonary disease (COPD), only a small proportion of smokers develop the disease. We tested the hypothesis that the response of the bronchial epithelium to cigarette smoke (CS) differs in patients with COPD. Such a difference might explain in part why only some cigarette smokers develop the disease. We established primary explant cultures of human bronchial epithelial cells (HBEC) from biopsy material obtained from never-smokers who had normal pulmonary function, smokers with normal pulmonary function, and smokers with COPD, and exposed these for 20 min to CS or air. Measurements were subsequently made over a period of 24 h of transepithelial permeability and release of interleukin (IL)-1beta and soluble intercellular adhesion molecule-1 (sICAM-1). In addition, intracellular reduced glutathione (GSH) levels were measured after 24 h incubation. Exposure to CS increased the permeability of these cultures in all study groups, but the most marked effect was observed in cultures from patients with COPD (mean increase, 85.5%). The smallest CS-induced increase in the permeability was observed in HBEC cultured from smokers with normal pulmonary function (mean, 25.0%), and this was significantly lower than that of HBEC from never-smokers (mean, 53.4%) (P<0.001). Compared with exposure to air, exposure to CS led to a significantly increased release of these mediators from cultures of the never-smoker group (mean 250.0% increase in IL-1beta and mean 175.3% increase in sICAM-1 24 h after exposure) and COPD group (mean 383.3% increase in IL-1beta and mean 97.4% increase in sICAM-1 24 h after exposure). In contrast, CS exposure did not influence significantly the release of either mediator from the cells of smokers with normal pulmonary function. Levels of intracellular GSH were significantly higher in cultures of HBEC derived from smokers, both those with normal pulmonary function and those with COPD, compared with cultures from healthy never-smokers. Exposure to CS significantly decreased the concentration of intracellular GSH in all cultures. However, the fall in intracellular GSH was significantly greater in cells from patients with COPD (mean 72.9% decrease) than in cells from never-smokers (mean 61.4% decrease; P = 0.048) or smokers with normal pulmonary function (mean 43.9% decrease; P = 0.02). These results suggest that whereas smokers with or without COPD demonstrate increased levels of GSH within bronchial epithelial cell cultures, those with COPD have a greater susceptibility to the effects of CS in reducing GSH levels and causing increased permeability and release of proinflammatory mediators such as IL-1beta and sICAM 1. PMID- 11017920 TI - Quantitative trait loci controlling allergen-induced airway hyperresponsiveness in inbred mice. AB - Identification of the genetic loci underlying asthma in humans has been hampered by variability in clinical phenotype, uncontrolled environmental influences, and genetic heterogeneity. To circumvent these complications, the genetic regulation of asthma-associated phenotypes was studied in a murine model. We characterized the strain distribution patterns for the asthma-related phenotypes airway hyperresponsiveness (AHR), lung eosinophils, and ovalbumin (OVA)-specific serum immunoglobulin (Ig) E induced by allergen exposure protocols in A/J, AKR/J, BALB/cJ, C3H/HeJ, and C57BL/6J inbred strains and in (C3H/HeJ x A/J)F1 mice. Expression of AHR differed between strains and was sometimes discordant with lung eosinophils or serum IgE. Furthermore, we identified two distinct quantitative trait loci (QTL) for susceptibility to allergen-induced AHR, Abhr1 (allergen induced bronchial hyperresponsiveness) (lod = 4. 2) and Abhr2 (lod = 3.7), on chromosome 2 in backcross progeny from A/J and C3H/HeJ mice. In addition, a QTL on chromosome 7 was suggestive of linkage to this trait. These QTL differ from those we have previously found to control noninflammatory AHR in the same crosses. Elucidation of the genes underlying these QTL will facilitate the identification of biochemical pathways regulating AHR in animal models of asthma and may provide insights into the pathogenesis of human disease. PMID- 11017921 TI - Mechanisms of proliferation synergy by receptor tyrosine kinase and G protein coupled receptor activation in human airway smooth muscle. AB - Despite recent studies depicting the capacity of G protein-coupled receptors (GPCRs) to activate mitogenic signaling pathways more commonly associated with receptor tyrosine kinases (RTKs), little is known regarding the interactive effects of GPCR and RTK activation on cell growth and signal transduction. Such interactions likely mediate the physiologic growth in most cells in vivo as well as the aberrant, non-neoplastic growth that occurs in diseases such as asthma, where disruptions of the local hormonal or inflammatory state can contribute to significant GPCR activation. In this study, we show that numerous inflammatory or contractile agents, including thrombin, histamine, and carbachol, potentiate epidermal growth factor (EGF)-stimulated proliferation of human airway smooth muscle (ASM), thus demonstrating a clear synergy between RTK and GPCR activation. Alterations in promitogenic nuclear signaling were evidenced by additive or synergistic increases in Elk-1 and activator protein-1 activation, and by increases in cyclin D1 expression. Interestingly, GPCR activation did not cause EGF receptor tyrosine phosphorylation nor did it increase EGF-stimulated autophosphorylation. In the presence of EGF, histamine or carbachol did not alter the time-dependent phosphorylation of p42/p44, whereas thrombin was capable of increasing phospho-p42/p44 levels at selected time points in some, but not all, cultures. In contrast to their relative inability to alter EGF receptor-linked p42/p44 activation, thrombin, histamine, and carbachol consistently increased the late phase (> 1 h) activity of p70 S6 kinase. Collectively, these findings suggest that inflammatory and contractile agents that activate GPCRs can significantly modulate RTK-mediated ASM growth through a p70 S6 kinase-dependent, p42/p44-independent mechanism. PMID- 11017922 TI - An antisense of protein kinase C-zeta inhibits proliferation of human airway smooth muscle cells. AB - We hypothesized that an atypical isoform of protein kinase (PK) C, PKC-zeta, is essential for proliferation of human airway smooth muscle (HASM) cells in primary culture. Recombinant replication-deficient E1-deleted adenoviruses (100 plaque forming units [pfu]/cell) expressing the antisense of PKC-zeta and the wild-type PKC-zeta (Ad-CMV-PKC-zeta) were added to actively growing cells that were subsequently incubated for 48 h in platelet-derived growth factor (PDGF) 40 ng/mL or 10% fetal bovine serum (FBS). Expression of the antisense at a virus concentration of 100 pfu/cell produced a significant (n = 3, P<0.05) decrease in the mean manual cell count in the presence of PDGF to 37+/-5% relative to that in cells with no virus (100%), whereas in cells infected with virus containing no construct, this figure was 102+/-13%. The increase in cell number in response to FBS, however, was not affected by the presence of the antisense. Corresponding values for cells in 10% FBS were 100+/-22%, 85+/-22%, and 122+/-18%. Western blotting revealed decreased levels of PKC-zeta protein, but not PKC-alpha or PKC epsilon protein, in cells infected with the antisense when compared with levels in control cells. Thus, in HASM cells, PKC-zeta is involved in proliferation in response to PDGF, but not in response to FBS, for which alternate signal transduction pathways independent of PKC-zeta must exist. PMID- 11017923 TI - Post-transcriptional deregulation of myc genes in lung cancer cell lines. AB - Genes of the myc family are frequently overexpressed in lung cancer. Gene amplification can explain the deregulation of these genes in a subset of tumors and cell lines, but in most cases, the cause of the elevated myc expression remains unknown. We examined whether messenger RNA (mRNA) stabilization could be contributing to myc gene overexpression in lung cancer cell lines. The decay pattern of c-myc or N-myc mRNA was analyzed in 11 such cell lines and in unimmortalized human embryonic lung cells. Eight lung cancer cell lines showed stabilization of c-myc or N-myc transcripts. To determine whether this stabilization was unique to myc genes, the decay pattern of the unstable c-fos proto-oncogene mRNA was also studied. The same cell lines that exhibited stabilization of myc mRNA showed an abnormally slow decay of the c-fos message, suggesting that there might be a correlation between the abnormal decay of c-fos and myc transcripts. In contrast, the half-life of histone 2B mRNA, which is degraded in a cell cycle-specific manner, did not appear to correlate with that of myc and fos. Our results suggest that an mRNA decay pathway responsible for the destruction of unstable proto-oncogene mRNAs may be commonly affected in lung cancers. PMID- 11017924 TI - The tyrosine kinases p53/56lyn and p72syk are differentially expressed at the protein level but not at the messenger RNA level in nonreleasing human basophils. AB - Within the general population, individuals can be found whose basophils do not secrete after stimulation through the immunoglobulin (Ig) E receptor. In this study we compared two groups of donors, those whose basophils responded with 65+/ 16% histamine release to an optimal concentration of anti-IgE antibody and those whose basophil response was not statistically different from nonstimulated release (1+/-1%). We show that these so-called nonreleasing basophils have at least 10-fold lower expression of the tyrosine kinases, lyn and syk, but normal expression of the tyrosine kinase Btk when compared with the panel of releasing basophils. Indeed, maximum histamine release correlated with expression of both syk (Spearman rank correlation coefficient [Rs] = 0.98) and lyn (Rs = 0.93). In contrast, equivalent levels of messenger RNA (mRNA) for lyn and syk kinase were found for both groups. By sequencing a critical region in the syk mRNA, our results also demonstrate that the frame shift mutation in syk leading to a premature stop codon which has been observed in other cell types is not present in nonreleasing human basophils. Our results suggest that there may be translational or post-translational regulatory mechanisms specific to the expression of two important FcepsilonRI-associated signaling elements in basophils. PMID- 11017925 TI - Priming of alveolar macrophages by leukotriene D(4): potentiation of inflammation. AB - Cysteinyl leukotrienes (LTs), including LTC(4), LTD(4), and LTE(4), are well known to induce bronchoconstriction and increase bronchial hyperreactivity, mucus secretion, and vascular permeability. Interestingly, alveolar macrophages (AMs) express LTD(4) high-affinity receptor. These cells represent a major source of inflammatory mediators implicated in the pathophysiology of asthma. Thus, we investigated the immunomodulatory effects of LTD(4) on the production of inflammatory mediators such as macrophage inflammatory protein (MIP)- 1alpha, tumor necrosis factor (TNF), and nitric oxide (NO) by AMs. NR8383 cells, an AM cell line, were pretreated with LTD(4) (10(-11) M) for different periods of time and stimulated or not with lipopolysaccharide (LPS) for 2 h. Although LTD(4) treatment did not modulate the release of MIP-1alpha and TNF, this treatment (6 h) significantly increased the release of these mediators when AMs were further stimulated with LPS (increases of 47 and 21%, respectively). Further, LTD(4) pretreatment increased messenger RNA (mRNA) levels of MIP-1alpha and TNF. These effects of LTD(4) were abrogated by the presence of a LTD(4) receptor antagonist, Verlukast (MK-679), showing the specificity of LTD(4). Interestingly, LTD(4) treatment significantly increased the release of NO by LPS-stimulated AMs without modulating mRNA levels of the inducible NO synthase. Our data suggest that LTD(4) primes AMs to release more MIP-1alpha, TNF, and NO after stimulation. Thus, in addition to its potent bronchoconstrictor effect, LTD(4) may participate in the inflammatory process seen in asthma by potentiating the production of proinflammatory mediators by AMs during immunologic stimuli. PMID- 11017926 TI - Sequencing with microarray technology--a powerful new tool for molecular diagnostics. PMID- 11017927 TI - Detection of prevalent genetic alterations predisposing to hemochromatosis and other common human diseases. PMID- 11017928 TI - Measurement of cardiac natriuretic hormones (atrial natriuretic peptide, brain natriuretic peptide, and related peptides) in clinical practice: the need for a new generation of immunoassay methods. AB - BACKGROUND: Cardiac natriuretic hormones (CNHs) are a family of related peptides, including atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and other peptides derived from the N-terminal portion of the proANP and proBNP peptide chains. Assays for cardiac natriuretic peptides have been proposed to help assess clinical conditions associated with expanded fluid volume. In particular, the assays can be useful for distinguishing healthy subjects from patients in different stages of heart failure. Measurements of these hormones have also been considered for prognostic indicators of long-term survival in patients with heart failure and/or after acute myocardial infarction. The different CNHs differ in their production/secretion patterns and have different clearance rates. Furthermore, there are numerous proposed assay configurations for each of these hormones, and it is not clear which assay provides the best pathophysiological and/or clinical information. APPROACH: Here we review recent studies concerning the competitive (such as RIA, enzyme immunoassay, or luminescence immunoassay) and noncompetitive immunoassays (such as two-site IRMA, ELISA, or immunoluminometric assay) for the different cardiac natriuretic peptides to compare the analytical characteristics and clinical relevance of assays for the different CNHs and the different assay formats. CONTENT: Developing sensitive, precise, and accurate immunoassays for cardiac natriuretic peptides has been difficult because of their low concentrations (on average, approximately 3-6 pmol/L) in healthy subjects and because of their structural, metabolic, and physiological characteristics. Competitive assays have historically suffered from lack of sensitivity and specificity for the biologically active peptides. These usually require tedious extraction procedures prior to analysis. Recently, immunometric assays have been developed that have improved sensitivity and specificity; it appears these will be the methods of choice. SUMMARY: To date, there is no consensus on the best assay procedure of cardiac natriuretic peptides. To facilitate widespread propagation of determination of these hormones in routine clinical practice, it will be necessary to study the new generation of noncompetitive immunometric methods that are less time-consuming and more sensitive and specific. Although several studies suggest that BNP exhibits better clinical utility than the other CNHs, more studies examining multiple CNHs in the same cohorts of patients will be necessary. PMID- 11017929 TI - Transferrin polymorphism influences iron status in blacks. AB - BACKGROUND: Genetic variants of human transferrin (TF) have been described, but little is known about their functional differences. We studied iron status according to TF phenotype in a healthy Zimbabwean population and in subjects at risk of African iron overload. METHODS: The study population consisted of 483 nondrinkers, 31 drinking spouse pairs, and 5 family pedigrees (n = 88) with index cases of iron overload. TF phenotypes were determined using starch gel electrophoresis. To evaluate iron status, serum iron, total iron-binding capacity (TIBC), ferritin, and soluble TF receptors were measured, and the percentage of saturation and the serum iron:TF ratio were calculated. The binding of the TF variants was studied by equilibrium dialysis. RESULTS: The reference population was characterized by a high TF D allele frequency (0.050) and a complete absence of homozygous TF DD individuals. Similar allele frequencies were observed in subjects at risk of African iron overload. In the reference population, male TF CD heterozygotes had significantly lower (P <0.01) values for serum iron, TIBC, TF saturation, and serum iron:TF ratio than the TF CC homozygotes; in females, only TIBC was significantly different. Overall red blood cell indices did not differ according to TF phenotype. In the population at risk of African iron overload, only serum iron:TF ratio was consistently significantly lower in TF CD phenotypes (P <0.05). After equilibrium dialysis, the amount of iron bound by TF was significantly lower (P <0.01) in TF CD individuals. CONCLUSIONS: The present data demonstrate a functional difference between TF phenotypes in blacks. PMID- 11017930 TI - Rapid single-tube screening of the C282Y hemochromatosis mutation by real-time multiplex allele-specific PCR without fluorescent probes. AB - BACKGROUND: An accurate determination of the major HFE mutation (C282Y), which is associated with hereditary hemochromatosis, is important in diagnosis and risk assessment for this disease. We report a single-tube high-throughput PCR method for the detection of C282Y. METHODS: We combined three previously described principles: allele-specific PCR, mutagenically separated PCR, and amplicon identification by specific dissociation curves. PCR amplification was performed with fluorescence detection or conventional thermocycler using the same primers, reactant constituents, and cycling protocol. Primer cross-reactions were prevented by deliberate primer:primer and primer:template mismatches. RESULTS: PCR products were identified by their characteristic melting temperatures based on SYBR Green I fluorescence. For each of the 256 random and 17 known HFE C282Y samples, mutant homozygous, wild-type, and heterozygous samples were unequivocally distinguished. CONCLUSIONS: This homogeneous assay is rapid, reproducible, does not require fluorescent oligonucleotide probes, and correctly identifies HFE genotypes. PMID- 11017931 TI - Genotypes and phenotypes for apolipoprotein E and Alzheimer disease in the Honolulu-Asia aging study. AB - BACKGROUND: The utility of apolipoprotein E (ApoE) type as an indicator of genetic susceptibility to Alzheimer disease (AD) depends on the reliability of typing. Although ApoE protein isoform phenotyping is generally assumed equivalent to genotyping from DNA, phenotype-genotype differences have been reported. METHODS: ApoE genotype and phenotype results were examined for 3564 older (ages 71-93 years) Japanese-American male participants of the Honolulu-Asia Aging Study, an ongoing population-based study of aging and dementia. RESULTS: Both methods demonstrated similar associations of ApoE type with AD: a direct association with ApoE4 and a less dramatic inverse association ApoE2. Advanced age did not appear to influence the ApoE4-AD association. The association with AD among ApoE4 homozygotes [odds ratio (OR) = 14.7] was higher than expected based on an observed OR of 2.0 in heterozygotes. Phenotype-genotype nonconcordance was more frequent for ApoE2 than for ApoE4. The ApoE2 phenotype occurred at a frequency of 7.9% vs a genotype frequency of 4.9%, corresponding to a probability of 56% that an individual with ApoE2 phenotype had the same genotype. CONCLUSIONS: Whereas E4 and E2 phenotypes and genotypes were comparably associated with AD, neither method would be expected to substantially improve the efficiency of case finding in the context of population screening beyond prediction based on age and education. Nonconcordance of phenotype and genotype was substantial for E2 and modest for E4 in this population. The ApoE4-AD association was independent of age. PMID- 11017932 TI - Evaluation of the performance of a p53 sequencing microarray chip using 140 previously sequenced bladder tumor samples. AB - BACKGROUND: Testing for mutations of the TP53 gene in tumors is a valuable predictor for disease outcome in certain cancers, but the time and cost of conventional sequencing limit its use. The present study compares traditional sequencing with the much faster microarray sequencing on a commercially available chip and describes a method to increase the specificity of the chip. METHODS: DNA from 140 human bladder tumors was extracted and subjected to a multiplex-PCR before loading onto the p53 GeneChip from Affymetrix. The same samples were previously sequenced by manual dideoxy sequencing. In addition, two cell lines with two different homozygous mutations at the TP53 gene locus were analyzed. RESULTS: Of 1464 gene chip positions, each of which corresponded to an analyzed nucleotide in the sequence, 251 had background signals that were not attributable to mutations, causing the specificity of mutation calling without mathematical correction to be low. This problem was solved by regarding each chip position as a separate entity with its own noise and threshold characteristics. The use of background plus 2 SD as the cutoff improved the specificity from 0.34 to 0.86 at the cost of a reduced sensitivity, from 0.92 to 0.84, leading to a much better concordance (92%) with results obtained by traditional sequencing. The chip method detected as little as 1% mutated DNA. CONCLUSIONS: Microarray-based sequencing is a novel option to assess TP53 mutations, representing a fast and inexpensive method compared with conventional sequencing. PMID- 11017933 TI - Enzymatic mutation detection in the P53 gene. AB - BACKGROUND: The enzymatic mutation detection (EMD) assay uses the bacteriophage resolvase T4 endonuclease VII, which cleaves preformed heteroduplex molecules at mismatch sites, forming two shorter fragments that can be resolved by gel electrophoresis. The method can be used to detect single and multiple base changes, as well as insertions and deletions. METHODS: The sensitivity, specificity, and positional accuracy of mutation detection by EMD with the PASSPORT(TM) Mutation Scanning Kit were assessed in a blind fashion for three analytical platforms (radioactive detection and automated laser sequencers ALFexpress and ABI PRISM 377). PCR products of 703 bp covering codons 188-393 of the P53 gene were prepared from colorectal tumor samples and analyzed by EMD; the results were compared to data from cDNA sequencing. A 1362-bp PCR product prepared from IL4r gene was used to test detection of multiple base changes in long PCR products. RESULTS: The sensitivity for detection of mutations using EMD exceeded 90%, and the specificity exceeded 80% on all analysis platforms. The method localized 90% of mutations to within two codons and four codons for automated laser sequencers and detection by radioactivity, respectively. The method detected at least five mismatches in heteroduplexes >1 kb. CONCLUSIONS: The EMD system facilitates efficient detection of genetic variation in fragments exceeding 1 kb irrespective of location and type. The technology is particularly well suited to the detection of mutations in genes frequently mutated at unpredictable locations. PMID- 11017934 TI - Rapid real-time fluorescent PCR gene dosage test for the diagnosis of DNA duplications and deletions. AB - BACKGROUND: Current methods to determine gene dosage are time-consuming and labor intensive. We describe a new and rapid method to assess gene copy number for identification of DNA duplications or deletions occurring in Charcot-Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP), respectively. METHODS: We studied 16 patients with HNPP, 4 with CMT1A, and 49 control subjects. We used real-time PCR on the LightCycler system with use of a single capillary tube and no post-PCR handling. A polymorphic fragment of the PMP22 gene was amplified to determine gene dosage for heterozygous samples. The presence of two alleles was used to indicate that no deletion was present in HNPP samples. The ratio obtained between the areas under each allele melting curve of heterozygous CMT1A samples was used to determine whether the sequence was duplicated or normal. Homozygous samples required a competitive gene dosage test, where the ratio between the areas under the melting curves of the target DNA of samples and of the competitor molecule was used to determine whether the target sequence was duplicated, deleted, or normal. Samples from HNPP, CMT1A, and controls were analyzed. RESULTS: Area ratios were approximately 0.6, 1.0, and 2.0 for HNPP, control, and CMT1A samples, respectively. The results agreed with those obtained by Southern blotting and microsatellite analysis in the same samples. CONCLUSIONS: Direct and competitive real-time fluorescent PCR can differentiate one, two, or three copies of the target DNA. The method described is sensitive and accurate for detection of CMT1A duplications and HNPP deletions and is faster and easier than current methods. PMID- 11017935 TI - Serum procalcitonin concentrations in term delivering mothers and their healthy offspring: a longitudinal study. AB - BACKGROUND: The reported sensitivities and specificities of procalcitonin (PCT) concentrations for the diagnosis of neonatal infection vary widely. A postnatal increase of PCT has been observed in healthy term newborns with a peak at approximately 24 h of age, and many questions remain regarding maternal and perinatal factors that may influence the normal PCT kinetics during the immediate postnatal period. METHODS: We prospectively investigated the association between the serum PCT values obtained from 121 mothers at delivery and serum PCT in their healthy, term offspring at birth as well as at 24 and 48 h of age. We also analyzed whether obstetric and perinatal factors would alter maternal and neonatal PCT response. RESULTS: PCT concentrations in the babies at birth were significantly higher than in the mothers (P <0.0001), with even larger differences at 24 and 48 h of age. None of the variables identified from maternal and perinatal histories had a significant effect on maternal PCT response. In the healthy neonate, the variables that significantly affected the concentration of PCT at birth were the mothers' PCT (P <0.01), maternal group B streptococcus colonization (P <0.05), and rupture of membranes >/=18 h (P <0.01). The coefficient of linear correlation between the mother's PCT concentration and that of the baby at birth was 0. 32 (P <0.01). The only variable that significantly altered the PCT concentration at both 24 (P <0.01) and 48 (P <0.01) h of age was rupture of membranes >/=18 h. Nonetheless, the PCT response observed during the 48-h period after birth among healthy babies born to mothers with risk factors for infection was well below that reported previously among age-matched neonates with sepsis. CONCLUSIONS: The postnatal increase of PCT observed in the healthy neonate with peak values at 24 h of age most likely represents endogenous synthesis. In estimating the sensitivities and specificities of PCT for diagnosis of sepsis throughout the initial 48 h of life, it is important to consider the normal PCT kinetics and the pattern(s) of PCT response in the healthy neonate. PMID- 11017936 TI - Discriminating capacity of indole markers in the diagnosis of carcinoid tumors. AB - BACKGROUND: We evaluated the discriminating capacity of the indole markers urinary 5-hydroxyindoleacetic acid (5-HIAA), urinary serotonin, and platelet serotonin in the diagnosis of carcinoid tumors. METHODS: Indole markers were measured in 688 patients with suspected carcinoid disease. The initial values of indole markers from patients in whom a carcinoid tumor was confirmed during follow-up (n = 98) were used for ROC analysis. Two groups served as reference populations. The first consisted of 45 healthy individuals ("healthy controls"). The second was a random sample of 40 patients, drawn from the 590 (688 minus 98) patients with carcinoid-like symptoms but without a carcinoid tumor ("clinically suspected patients"). RESULTS: ROC curve analysis showed platelet serotonin to have the highest discriminating capacity, especially in foregut carcinoids. Cutoff values for platelet serotonin obtained from ROC analysis with healthy controls as reference group (5.4 nmol/10(9) platelets) gave a sensitivity of 74%, specificity of 91%, positive predictive value of 63%, and negative predictive value of 95% when applied to the initial 688 patients. Using the cutoff value with the clinically suspected patients as the reference group (9.3 nmol/10(9) platelets) gave a sensitivity of 63%, specificity of 99%, positive predictive value of 89%, and negative predictive value of 93%. Indole markers were increased in 169 (25%) of 688 patients. In 76 (45%) of these 169 patients, a carcinoid tumor was present. Slight increases of markers were associated with non-carcinoid neuroendocrine tumors, non-neuroendocrine tumors, and disturbed bowel motility. CONCLUSIONS: ROC curve analysis shows that platelet serotonin is the most discriminating indole marker for the diagnosis of carcinoid tumors. Platelet serotonin especially improves the diagnosis of carcinoids producing small amounts of serotonin. PMID- 11017937 TI - C-Reactive protein and cardiac troponin T in risk stratification: differences in optimal timing of tests early after the onset of chest pain. AB - BACKGROUND: Increased C-reactive protein (CRP) is an important prognostic indicator for early risk stratification in patients with an acute coronary syndrome (ACS), independent of, and in combination with, increased cardiac troponin T (cTnT). However, increases in both cTnT and CRP also occur secondary to myocardial damage. METHODS AND RESULTS: In 156 consecutive patients, early release kinetics of CRP and cTnT were analyzed. The cutoff values were 3.0 mg/L for CRP and 0.1 microgram/L for cTnT. In the 75 patients with a CRP below the cutoff on admission, there was little change in CRP until 8 h after the onset of symptoms. At 12 h after the onset of symptoms, the cumulative proportions of abnormal CRP and cTnT in non-ST elevation ACS patients were 27% and 89%, respectively (P <0.01). During the first 24 h after the onset of symptoms, the median time above the cutoff was 20 h for CRP and 5 h for cTnT (P <0.0001). CRP was below the cutoff on admission significantly more often among patients receiving thrombolytic therapy than in patients without an indication for reperfusion therapy (51% vs 28%; P = 0.004). CONCLUSIONS: Increased CRP as an early independent risk indicator should be measured as soon as possible after the onset of symptoms, whereas increased cTnT is most reliable at 12 or more hours after the onset of symptoms. PMID- 11017938 TI - Clinical evaluation of the first medical whole blood, point-of-care testing device for detection of myocardial infarction. AB - BACKGROUND: Validation of whole blood, point-of-care testing devices for monitoring cardiac markers to aid clinicians in ruling in and ruling out myocardial infarction (MI) is necessary for both laboratory and clinical acceptance. METHODS: This study evaluated the clinical diagnostic sensitivity and specificity of the First Medical Cardiac Test device operated by nursing and laboratory personnel that simultaneously measures cardiac troponin I (cTnI), creatine kinase (CK) MB, myoglobin, and total CK on the Alpha Dx analyzer in whole blood for detection of MI. Over a 6-month period, 369 patients initially presenting to the emergency department with chest pain were evaluated for MI using modified WHO criteria. Eighty-nine patients (24%) were diagnosed with MI. RESULTS: In whole blood samples collected at admission and at 3- to 6-h intervals over 24 h, ROC curve-determined MI decision limits were as follows: cTnI, 0.4 microgram/L; CKMB, 7.0 microgram/L; myoglobin, 180 microgram/L; total CK, 190 microgram/L. Based on peak concentrations within 24 h after presentation, the following sensitivities (+/- 95% confidence intervals) were found: cTnI, 93% +/- 5.5%; myoglobin, 81% +/- 9.7%; CKMB, 90% +/- 6.3%; total CK, 86% +/- 7.5%. Sensitivities were maximal at >90% for both cTnI and CKMB at >12 h in MI patients, without differences between ST-segment elevation and non-ST-segment elevation MI patients. CONCLUSIONS: The First Medical point-of-care device provides cardiac marker assays that can be used by laboratories and clinicians in a variety of hospital settings for ruling in and ruling out MI. PMID- 11017939 TI - Production and characterization of novel anti-prostate-specific antigen (PSA) monoclonal antibodies that do not detect internally cleaved Lys145-Lys146 inactive PSA. AB - BACKGROUND: The nature of free, uncomplexed prostate-specific antigen (PSA) in the circulation is still unknown. In this study, we developed novel anti-PSA antibodies using PSA produced by a metastasized cancer cell line, LNCaP, as an immunogen. METHODS: Hybridoma cell lines were screened with different methods that aimed at finding antibodies specific for the forms of free PSA produced by LNCaP cell line. Obtained antibodies were further studied for their characteristics related to previously characterized monoclonal antibodies. RESULTS: Numerous anti-PSA antibodies were obtained, of which four represented unique epitopes previously unrecognized by us. One free-PSA-specific antibody was bound to PSA on two distinct epitopes, and one antibody was bound to the carboxyl terminal peptide of PSA. Two antibodies were found to bind to the peptide sequence adjacent to the internal cleavage site Lys145-Lys146. These antibodies failed to recognize internally cleaved PSA at Lys145-Lys146. We could not find anti-proPSA antibodies despite the fact that LNCaP PSA contained more than one half of the zymogen form of PSA. CONCLUSIONS: We report, for the first time, novel anti-PSA antibodies that do not recognize internally cleaved PSA at Lys145 Lys146 and thus are specific for intact, unclipped PSA. PMID- 11017940 TI - The haptoglobin 2-2 phenotype affects serum markers of iron status in healthy males. AB - BACKGROUND: Human iron status is influenced by environmental and genetic factors. We hypothesized that the genetic polymorphism of haptoglobin (Hp), a hemoglobin binding plasma protein, could affect iron status. METHODS: Reference values of serum iron status markers were compared according to Hp phenotypes (Hp 1-1, Hp 2 1, Hp 2-2; determined by starch gel electrophoresis) in 717 healthy adults. Iron storage was investigated in peripheral blood monocyte-macrophages by measuring cytosolic L- and H-ferritins and by in vitro uptake of radiolabeled ((125)I) hemoglobin-haptoglobin complexes. RESULTS: In males but not in females, the Hp 2 2 phenotype was associated with higher serum iron (P <0.05), transferrin saturation (P <0.05), and ferritin (P <0.01) concentrations than Hp 1-1 and 2-1, whereas soluble transferrin receptor concentrations were lower (P <0.05). Moreover, serum ferritin correlated with monocyte L-ferritin content (r = 0.699), which was also highest in the male Hp 2-2 subgroup (P <0.01). In vitro, monocyte macrophages took up a small fraction of (125)I-labeled hemoglobin complexed to Hp 2-2 but not to Hp 1-1 or 2-1. CONCLUSIONS: The Hp 2-2 phenotype affects serum iron status markers in healthy males and is associated with higher L-ferritin concentrations in monocyte-macrophages because of a yet undescribed iron delocalization pathway, selectively occurring in Hp 2-2 subjects. PMID- 11017941 TI - Plasma nitric oxide concentrations and nitric oxide synthase gene polymorphisms in coronary artery disease. AB - BACKGROUND: Plasma NOx (nitrate and nitrite) is a stable end product of the vasodilator NO. Several polymorphisms in the endothelial constitutive NO synthase (ecNOS) gene have been reported, including the 4a/4b VNTR polymorphism in intron 4, the E298D mutation in exon 7, and the G10-T polymorphism in intron 23. The aims of this study were to examine plasma NOx in patients with coronary artery disease (CAD) and to assess the association between plasma NOx concentrations and the three ecNOS gene polymorphisms. METHODS: Plasma NOx was measured in samples from 128 healthy controls and from 110 CAD patients at least 2 months after myocardial infarction. Three genetic polymorphisms that are known or have been suggested to be associated with plasma NOx concentration were also analyzed by PCR-restriction fragment length polymorphism. RESULTS: Median plasma NOx was significantly higher (P <0.001) in CAD patients (95.9 micromol/L) than in controls (73.8 micromol/L). Furthermore, the median plasma NOx was significantly higher (P <0.001) in hypertensive CAD patients (116.0 micromol/L) than in controls and normotensive CAD patients (86.0 micromol/L). The G-allele frequency of the G10-T polymorphism in intron 23 was significantly higher in CAD patients than in controls. Other polymorphisms showed no differences in allelic frequencies among the control and CAD groups. In controls, individuals with the E298D mutation in exon 7 (136.1 micromol/L) showed significantly higher (P = 0.001) median plasma NOx than those without this mutation (64.5 micromol/L). CONCLUSIONS: Plasma NOx was higher in hypertensive CAD patients than in normotensive CAD patients and controls. The E298D polymorphism of the ecNOS gene was associated with increased plasma NOx. Further study is needed to understand the gene expression and enzyme activity of ecNOS and their association with genotypes. PMID- 11017942 TI - Multicenter evaluation of five assays for myoglobin determination. AB - BACKGROUND: Lacking assay standardization, different myoglobin methods may produce results that differ significantly. METHODS: A multicenter study was carried out to compare the analytical performance of five commercially available assays for myoglobin measurement. Linearity, imprecision, interferences, and method comparison were studied according to NCCLS guidelines, whereas reference values were determined following IFCC recommendations. RESULTS: The BNA and Opus showed relatively high imprecision (all but one total CV >7.4%). Other assays showed lower CVs, but they varied among laboratories, particularly at a normal myoglobin concentration (Access, 6.0-11%; Hitachi, 3.8-5.8%; Stratus, 3.4-6.5%). Results were lower in anticoagulated samples on the Access, in heparin and citrate samples on the Stratus, and in citrate samples on the BNA and Opus, and increased in heparin and EDTA samples on the Hitachi. Use of separator gel produced results significantly lower (P <0.001) on the Hitachi and higher (P = 0.016) on the Opus. Bilirubin, turbidity, and hemoglobin had no effect on evaluated methods, but rheumatoid factor affected the Access. In method comparisons, high correlation coefficients (>/=0.98) were obtained. The Stratus gave higher results; however, the Access and BNA gave the lowest. The following upper reference limits (microgram/L) for men and women, respectively, were obtained: Access, 70 and 52; BNA, 51 and 49; Hitachi, 67 and 58; Opus, 80 and 50; and Stratus, 86 and 63. CONCLUSION: The possibility of high imprecision and marked disagreement among commercial myoglobin assays should be carefully considered in clinical practice. PMID- 11017943 TI - Determination of apolipoprotein B-48 in plasma by a competitive ELISA. AB - BACKGROUND: Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as part of chylomicrons, and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk. Our aim was to develop, for routine analysis, an assay to quantify apoB-48 in plasma samples. METHODS: A microtiter plate was coated with a C-terminal apoB-48-specific heptapeptide. Plasma samples were incubated with appropriate detergent to allow competition between immobilized antigen and plasma apoB-48. Appropriate calibration curves were obtained in the ELISA, using calibrated lymph and chylomicrons. RESULTS: Treatment of plasma samples with the mild detergent Triton X-100 allowed an efficient competition between immobilized antigen and plasma apoB-48. No cross-reactivity was found with apoB-100, as checked by ELISA and Western blot analysis. Intra- and interassay CVs were 5.4% and 5. 5%, respectively. In healthy subjects, apoB-48 concentrations markedly increased in the postprandial state, in parallel with triglycerides. CONCLUSIONS: This new ELISA allows determination of the concentration of apoB-48 in normolipidemic plasma. PMID- 11017944 TI - Measurement of transcobalamin by ELISA. AB - BACKGROUND: Transcobalamin is essential for the cellular internalization of cobalamin. Methods to quantify the unsaturated protein are available, but few attempts have been made to develop methods to quantify the sum of unsaturated and cobalamin saturated transcobalamin. METHODS: gamma-Globulins from two polyclonal rabbit antibodies against recombinant human transcobalamin were used as capture and detection antibodies, and recombinant human transcobalamin was used as calibrator in an ELISA design. RESULTS: The ELISA is specific for transcobalamin and has a detection limit of <1.6 pmol/L. The imprecision (CV) is 4-6% for mean concentrations of 13-70 pmol/L. The central 95% interval for serum from healthy blood donors (n = 77) was approximately 600-1500 pmol/L and showed limited variation with age and sex. No correlation was observed between the marker of acute phase reaction, C-reactive protein, and transcobalamin in plasma. CONCLUSIONS: The ELISA measures total transcobalamin in serum and thus can be used for measurement of transcobalamin in patients treated with cobalamin. PMID- 11017945 TI - Determination of S-adenosylmethionine and S-adenosylhomocysteine in plasma and cerebrospinal fluid by stable-isotope dilution tandem mass spectrometry. AB - BACKGROUND: Available methods for the determination of nanomolar concentrations of S:-adenosylmethionine (SAM) and S:-adenosylhomocysteine (SAH) in plasma and cerebrospinal fluid (CSF) are time-consuming. We wished to develop a method for their rapid and simultaneous measurement. METHODS: We used tandem mass spectrometry (MS/MS) for the simultaneous determination of SAM and SAH, with stable-isotope-labeled internal standards. The (13)C(5)-SAH internal standard was enzymatically prepared using SAH-hydrolase and [(13)C(5)]adenosine. The method comprises a weak anion-exchange solid-phase extraction procedure serving as clean up step for the deproteinized plasma and CSF samples. After clean-up, samples were injected on a C(18) HPLC column, which was connected directly to the tandem mass spectrometer, operating in MS/MS mode. RESULTS: In plasma samples, the intraassay CVs for SAM and SAH were 4.2% and 4.0%, respectively, and the interassay CVs were 7.6% and 5. 9%, respectively. In CSF, the intraassay CVs for SAM and SAH were 6. 8% and 6.9%, respectively, and the interassay CVs were 4.2% and 5.5%, respectively. Mean recovery of SAM and SAH for both matrices at two concentrations was 93%. Detection limits for SAM and SAH in samples were 7.5 and 2.5 nmol/L, respectively. Concentrations of SAM and SAH in plasma from healthy subjects were within the previously reported ranges. In 10 CSF samples, the mean concentrations (range) were 248 (137-385) nmol/L for SAM and 11.3 (8.9-14.1) nmol/L for SAH. CONCLUSIONS: SAM and SAH can be analyzed by MS/MS, taking optimal advantage of the speed and high sensitivity and specificity of this relatively new analytical technique. PMID- 11017946 TI - Comparison of commercially available (125)I-based RIA methods for the determination of circulating 25-hydroxyvitamin D. AB - BACKGROUND: Measurement of circulating 25-hydroxyvitamin D [25(OH)D] is important in the management of metabolic bone disease. The aim of this study was to compare two widely used methods for the quantification of circulating 25(OH)D with attention to their abilities to measure 25-hydroxylated ergocalciferol (vitamin D(2)) [25(OH)D(2)] and cholecalciferol (vitamin D(3)) [25(OH)D(3)]. METHODS: We used two commercially available, Food and Drug Administration-approved, radioiodine ((125)I)-based RIA kits for the detection of 25(OH)D (DiaSorin, Stillwater, MN and IDS Ltd, Tyne and Wear, United Kingdom). These methods were tested for general assay performance, including antibody specificity. Results were compared with those of an HPLC-based direct ultraviolet detection method. RESULTS: Within- and between-run CVs were 95% for determination of anti-Scl70 antibodies. PMID- 11017950 TI - Total-homocysteine enzymatic assay. PMID- 11017951 TI - Effect of hemoglobin variants on routine glycohemoglobin measurements assessed by a mass spectrometric method. PMID- 11017952 TI - Simplified multiplex-PCR diagnosis of common southeast asian deletional determinants of alpha-thalassemia. PMID- 11017953 TI - Serum creatinine and fat-free mass (lean body mass). PMID- 11017955 TI - Identification of two distinct mutations at the same nucleotide position, concomitantly with a novel polymorphism in the vasopressin-neurophysin II gene (AVP-NP II) in two dutch families with familial neurohypophyseal diabetes insipidus. PMID- 11017954 TI - Rapid detection of the two most common CLN2 mutations causing classical late infantile neuronal ceroid lipofuscinosis. PMID- 11017956 TI - A sensitive assay of tumor necrosis factor alpha in sera from Duchenne muscular dystrophy patients. PMID- 11017957 TI - A golden age of clinical chemistry: 1948-1960. AB - This segment of history aims to inform the new, and remind the not-so-new, members of the profession about the relatively recent period that initiated the dominant role played by technological innovation in the modern investigation of disease. The 12 years from 1948 to 1960 were notable for introduction of the Vacutainer tube, electrophoresis, radioimmunoassay, and the AutoAnalyzer. Also appearing during this interval were new organizations, publications, programs, and services that established a firm foundation for the professional status of clinical chemists. It was a golden age. PMID- 11017958 TI - State of the art instead of biological variation to set requirements for imprecision. PMID- 11017959 TI - Evaluation of a simple dot-blot method for the detection of anti-neutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase. PMID- 11017960 TI - Dilution protocols for detection of hook effects/prozone phenomenon. PMID- 11017961 TI - Editor's pick PMID- 11017962 TI - This month in wjm PMID- 11017963 TI - The epidemic of pediatric obesity. PMID- 11017964 TI - Direct-to-consumer advertising of prescription drugs. PMID- 11017965 TI - Guidelines for managing acute bacterial meningitis in adults PMID- 11017966 TI - A book that made me think PMID- 11017967 TI - Should physicians be required to report domestic violence to the police? Against: mandatory reporting does not guarantee safety. PMID- 11017968 TI - Should physicians be required to report domestic violence to the police? For: mandatory reporting is worthy of support. PMID- 11017970 TI - Elizabeth hurley sucks to stop smoking PMID- 11017969 TI - Women physicians earn $63,000 less than male counterparts. PMID- 11017971 TI - Lumpectomy as good as mastectomy for tumors up to 5 cm across. PMID- 11017973 TI - Islet cell transplantation may be feasible PMID- 11017972 TI - Global news roundup PMID- 11017974 TI - $1 billion drug deal creates debt for "tomorrow's AIDS orphans". PMID- 11017975 TI - SAMe targets consumers via the Web. PMID- 11017977 TI - How to submit your medical images for publication in Med.Pix PMID- 11017976 TI - What is in a name? PMID- 11017978 TI - Preventing dog bites in children: randomized controlled trial of an educational intervention. PMID- 11017979 TI - Bark, don't bite! PMID- 11017980 TI - Grievances against physicians: 11 years' experience of a medical society grievance committee. AB - OBJECTIVE: To understand causes of patient dissatisfaction that result in complaints. DESIGN: Grievances received by the grievance committee between January 1, 1989, and January 1, 2000, were reviewed. SETTING: A 2-county area of North Carolina. SUBJECTS: Of 29 patients who filed grievances, the 9 male (31%) and 20 female (69%) patients had a mean (+/-SD) age of 39 (+/-19) years. In 18 instances, the patient consulted the physician less than 3 times (64%) before the complaint and in 8 instances more than 4 times (29%). Main outcome measures Allegations of the grievance and the committee's findings. RESULTS: Grievances fell into 5 categories: failure to fulfill expectations for examination and treatment (38%), failure to promptly diagnose (20%), rudeness (17%), producing excessive pain or practicing beyond the area of expertise (13%), and inappropriate behavior related to billings (10%). In 45% of the grievances, the committee found no breach of practice standards. In 17% of the cases, the physician resolved the grievance by apologizing, adjusting a bill, or completing insurance forms. CONCLUSION: Most grievances were filed by younger women against newly encountered physicians and were related to inadequate communication or alleged delay in diagnosis. PMID- 11017981 TI - Patient grievances are a continuing challenge for medical science. PMID- 11017982 TI - Recruitment and retention in the Navajo Area Indian Health Service. AB - OBJECTIVE: To determine why physicians and midlevel providers join, leave, or stay in Navajo Area Indian Health Service (IHS). DESIGN: Cross-sectional analysis of data obtained from questionnaires. SETTING: Navajo Area IHS hospitals. SUBJECTS: Navajo Area health care physicians and midlevel health care providers. Main outcome measures The prevalence of physicians and midlevel providers who plan to leave the Navajo Area IHS, the demographic characteristics of these physicians and midlevel providers, and the most common reasons for staying or leaving. RESULTS: A total of 221 (64%) physicians and midlevel providers responded. Of these, 58% planned to leave eventually, and 47% of all physicians and midlevel providers planned to leave in the next 3 years. Physicians and midlevel providers planning to leave tended to be younger than those planning to stay (P: = 0.009). The most common reason to join the IHS was a desire to work in the Southwest, to stay was the quality of the medical staff, and to leave was lack of administrative support. CONCLUSIONS: A high turnover rate of physicians and midlevel providers may occur in the next 3 years. A combination of factors specific to the provider, the institution, and the environment attracts physicians to the Navajo IHS and encourages them to stay. Factors that push physicians and midlevel providers to leave tend to be specific to the institution and are potentially amenable. PMID- 11017983 TI - How long will you be staying, doctor? PMID- 11017984 TI - Medical malpractice: the effect of doctor-patient relations on medical patient perceptions and malpractice intentions. AB - OBJECTIVE: To examine the causal effects of doctor-patient relations and the severity of a medical outcome on medical patient perceptions and malpractice intentions in the event of an adverse medical outcome. DESIGN: Randomized between subjects experimental design. Patients were given scenarios depicting interactions between an obstetric patient and her physician throughout the patient's pregnancy, labor, and delivery. PARTICIPANTS: One hundred twenty-eight postpartum obstetric patients were approached for participation, of whom 104 completed the study. Main outcome measures Patients' perceptions of physician competence and intentions to file a malpractice claim. RESULTS: Positive physician communication behaviors increased patients' perceptions of physician competence and decreased malpractice claim intentions toward both the physician and the hospital. A more severe outcome increased only patients' intentions to sue the hospital. CONCLUSION: These results provide empiric evidence for a direct, causal effect of the doctor-patient relationship on medical patients' treatment perceptions and malpractice claim intentions in the event of an adverse medical outcome. PMID- 11017986 TI - Commentary PMID- 11017985 TI - Communication: it is common sense. PMID- 11017987 TI - Is a clinical prediction model accurate for predicting ectopic pregnancy? PMID- 11017988 TI - Commentary PMID- 11017989 TI - Is hyaluronic acid effective in patients with osteoarthritis of the knee? PMID- 11017990 TI - Commentary PMID- 11017992 TI - Secondary prevention of transient ischemic attack and stroke PMID- 11017991 TI - Are spacers made from sealed cold-drink bottles as effective as conventional spacers? PMID- 11017993 TI - Treating hypertension. PMID- 11017994 TI - Notifying survivors about sudden, unexpected deaths. PMID- 11017995 TI - Bell's palsy. PMID- 11017996 TI - Vaccination updates. PMID- 11017997 TI - Netphiles PMID- 11017998 TI - Exercise for cancer patients: a new challenge in sports medicine PMID- 11017999 TI - Are phytoestrogens a "natural alternative" to estrogen replacement therapy? PMID- 11018000 TI - When should physicians forgo curative treatment of pneumonia in patients with dementia? Using a guideline for decision-making. PMID- 11018001 TI - Pneumonia, friend of the elderly. PMID- 11018002 TI - Filipino attitudes toward pain medication. A lesson in cross-cultural care. PMID- 11018004 TI - Dying people still want to care for others PMID- 11018003 TI - Physician-patient relationship: like marriage, without the romance. PMID- 11018005 TI - Multivitamins should be taken early in pregnancy PMID- 11018006 TI - Any Questions? any answers? PMID- 11018007 TI - Myth: codeine is an effective cough suppressant for upper respiratory tract infections. PMID- 11018008 TI - Cholera PMID- 11018009 TI - The Rb/E2F pathway: expanding roles and emerging paradigms. PMID- 11018010 TI - Molecular genetics of prostate cancer. PMID- 11018011 TI - Dynamic association of capping enzymes with transcribing RNA polymerase II. AB - The C-terminal heptad repeat domain (CTD) of RNA polymerase II (pol II) is proposed to target pre-mRNA processing enzymes to nascent pol II transcripts, but this idea has not been directly tested in vivo. In vitro, the yeast mRNA capping enzymes Ceg1 and Abd1 bind specifically to the phosphorylated CTD. Here we show that yeast capping enzymes cross-link in vivo to the 5' ends of transcribed genes and that this localization requires the CTD. Both the extent of CTD phosphorylation at Ser 5 of the heptad repeat and the binding of capping enzymes decreased as polymerase moved from the 5' to the 3' ends of the ACT1, ENO2, TEF1, GAL1, and GAL10 genes. Ceg1 is released early in elongation, but Abd1 can travel with transcribing pol II as far as the 3' end of a gene. The CTD kinase, Kin28, is required for binding, and the CTD phosphatase, Fcp1, is required for dissociation of capping enzymes from the elongation complex. CTD phosphorylation and dephosphorylation therefore control the association of capping enzymes with pol II as it transcribes a gene. PMID- 11018012 TI - Functional selectivity of recombinant mammalian SWI/SNF subunits. AB - The SWI/SNF family of chromatin-remodeling complexes plays a key role in facilitating the binding of specific transcription factors to nucleosomal DNA in diverse organisms from yeast to man. Yet the process by which SWI/SNF and other chromatin-remodeling complexes activate specific subsets of genes is poorly understood. We show that mammalian SWI/SNF regulates transcription from chromatin assembled genes in a factor-specific manner in vitro. The DNA-binding domains (DBDs) of several zinc finger proteins, including EKLF, interact directly with SWI/SNF to generate DNase I hypersensitivity within the chromatin-assembled beta globin promoter. Interestingly, we find that two SWI/SNF subunits (BRG1 and BAF155) are necessary and sufficient for targeted chromatin remodeling and transcriptional activation by EKLF in vitro. Remodeling is achieved with only the BRG1-BAF155 minimal complex and the EKLF zinc finger DBD, whereas transcription requires, in addition, an activation domain. In contrast, the BRG1-BAF155 complex does not interact or function with two unrelated transcription factors, TFE3 and NF-kappaB. We conclude that specific domains of certain transcription factors differentially target SWI/SNF complexes to chromatin in a gene-selective manner and that individual SWI/SNF subunits play unique roles in transcription factor directed nucleosome remodeling. PMID- 11018013 TI - Different phosphorylated forms of RNA polymerase II and associated mRNA processing factors during transcription. AB - The activities of several mRNA processing factors are coupled to transcription through binding to RNA polymerase II (Pol II). The largest subunit of Pol II contains a repetitive carboxy-terminal domain (CTD) that becomes highly phosphorylated during transcription. mRNA-capping enzyme binds only to phosphorylated CTD, whereas other processing factors may bind to both phosphorylated and unphosphorylated forms. Capping occurs soon after transcription initiation and before other processing events, raising the question of whether capping components remain associated with the transcription complex after they have modified the 5' end of the mRNA. Chromatin immunoprecipitation in Saccharomyces cerevisiae shows that capping enzyme cross-links to promoters but not coding regions. In contrast, the mRNA cap methyltransferase and the Hrp1/CFIB polyadenylation factor cross-link to both promoter and coding regions. Remarkably, the phosphorylation pattern of the CTD changes during transcription. Ser 5 phosphorylation is detected primarily at promoter regions dependent on TFIIH. In contrast, Ser 2 phosphorylation is seen only in coding regions. These results suggest a dynamic association of mRNA processing factors with differently modified forms of the polymerase throughout the transcription cycle. PMID- 11018014 TI - A Drosophila IkappaB kinase complex required for Relish cleavage and antibacterial immunity. AB - Here we report the identification of a Drosophila IkappaB kinase complex containing DmIKKbeta and DmIKKgamma, homologs of the human IKKbeta and IKKgamma proteins. We show that this complex is required for the signal-dependent cleavage of Relish, a member of the Rel family of transcriptional activator proteins, and for the activation of antibacterial immune response genes. In addition, we find that the activated DmIKK complex, as well as recombinant DmIKKbeta, can phosphorylate Relish in vitro. Thus, we propose that the Drosophila IkappaB kinase complex functions, at least in part, by inducing the proteolytic cleavage of Relish. The N terminus of Relish then translocates to the nucleus and activates the transcription of antibacterial immune response genes. Remarkably, this Drosophila IkappaB kinase complex is not required for the activation of the Rel proteins Dif and Dorsal through the Toll signaling pathway, which is essential for antifungal immunity and dorsoventral patterning during early development. Thus, a yet to be identified IkappaB kinase complex must be required for Rel protein activation via the Toll signaling pathway. PMID- 11018015 TI - The forkhead domain gene unc-130 generates chemosensory neuron diversity in C. elegans. AB - Caenorhabditis elegans responds to its complex chemical environment using a small number of chemosensory neurons. Each of these neurons exhibits a unique sensory response repertoire. The developmental mechanisms that generate this diversity of function are largely unknown. Many C. elegans chemosensory neurons, including the AWA and ASG neurons, arise as lineal sisters of an asymmetric division. Here we describe the gene unc-130, which plays a role in the generation of the AWA and ASG neurons. In unc-130 mutants, the ASG neurons adopt the fate of the AWA neurons. unc-130 encodes a member of the forkhead domain family of transcription factors, and is expressed in the precursors to AWA and ASG neurons. Misexpression of unc-130 in the AWA neurons is partly sufficient to repress the AWA fate, but not to promote ASG fate. unc-130 also plays a role in the development of additional chemosensory neurons. Our experiments show that the ASG neurons share a developmental default state in common with three types of olfactory neurons. We propose that distinct cell fates and hence diversity of function in the chemosensory neurons of C. elegans are generated in a hierarchical manner, utilizing both lineage-dependent and independent mechanisms. PMID- 11018016 TI - The forkhead transcription factor UNC-130 is required for the graded spatial expression of the UNC-129 TGF-beta guidance factor in C. elegans. AB - Secreted proteins required for cellular movements along the circumference of the body wall in Caenorhabditis elegans include UNC-6/netrin and the novel TGF-beta UNC-129. Expression of these proteins is graded along the dorsoventral (D/V) axis, providing polarity information to guide migrations. Here we show that the graded expression of UNC-129 in dorsal but not ventral body muscles depends on unc-130, which encodes a Forkhead transcription factor. The phenotype of unc-130 mutants closely mimics the reported effects of ectopically expressing unc-129 in both dorsal and ventral body muscles (). This fits our present finding that unc 130 cell autonomously represses unc-129 expression in the ventral body muscles. Thus the cell-specific effects of unc-130 on ventral, but not dorsal, body muscle expression of unc-129 accounts for the D/V polarity information required for UNC 129-mediated guidance. Genetic interactions between unc-130 and other guidance genes show that several molecular pathways function in parallel to guide the ventral to dorsal migration of distal tip cells (DTCs) and axonal growth cones in C. elegans. Genetic interactions confirm that UNC-129 does not require the only known type II TGF-beta receptor in C. elegans (DAF-4) for its guidance functions. Also, unc-130 is partially required for male tail morphogenesis and for embryogenesis. PMID- 11018017 TI - Multiple Ras-dependent phosphorylation pathways regulate Myc protein stability. AB - Our recent work has shown that activation of the Ras/Raf/ERK pathway extends the half-life of the Myc protein and thus enhances the accumulation of Myc activity. We have extended these observations by investigating two N-terminal phosphorylation sites in Myc, Thr 58 and Ser 62, which are known to be regulated by mitogen stimulation. We now show that the phosphorylation of these two residues is critical for determining the stability of Myc. Phosphorylation of Ser 62 is required for Ras-induced stabilization of Myc, likely mediated through the action of ERK. Conversely, phosphorylation of Thr 58, likely mediated by GSK-3 but dependent on the prior phosphorylation of Ser 62, is associated with degradation of Myc. Further analysis demonstrates that the Ras-dependent PI-3K pathway is also critical for controlling Myc protein accumulation, likely through the control of GSK-3 activity. These observations thus define a synergistic role for multiple Ras-mediated phosphorylation pathways in the control of Myc protein accumulation during the initial stage of cell proliferation. PMID- 11018018 TI - Antagonism between C/EBPbeta and FOG in eosinophil lineage commitment of multipotent hematopoietic progenitors. AB - The commitment of multipotent cells to particular developmental pathways requires specific changes in their transcription factor complement to generate the patterns of gene expression characteristic of specialized cell types. We have studied the role of the GATA cofactor Friend of GATA (FOG) in the differentiation of avian multipotent hematopoietic progenitors. We found that multipotent cells express high levels of FOG mRNA, which were rapidly down-regulated upon their C/EBPbeta-mediated commitment to the eosinophil lineage. Expression of FOG in eosinophils led to a loss of eosinophil markers and the acquisition of a multipotent phenotype, and constitutive expression of FOG in multipotent progenitors blocked activation of eosinophil-specific gene expression by C/EBPbeta. Our results show that FOG is a repressor of the eosinophil lineage, and that C/EBP-mediated down-regulation of FOG is a critical step in eosinophil lineage commitment. Furthermore, our results indicate that maintenance of a multipotent state in hematopoiesis is achieved through cooperation between FOG and GATA-1. We present a model in which C/EBPbeta induces eosinophil differentiation by the coordinate direct activation of eosinophil-specific promoters and the removal of FOG, a promoter of multipotency as well as a repressor of eosinophil gene expression. PMID- 11018019 TI - Unphosphorylatable mutants of Cdc6 disrupt its nuclear export but still support DNA replication once per cell cycle. AB - Cdc6 is essential for eukaryotic DNA replication. We have mutated highly conserved CDK phosphorylation sites in Cdc6. Contrary to their reported phenotypes in human cells, unphosphorylatable DeltaCDK mutants fully support DNA replication in Xenopus eggs. WtCdc6 is actively exported from the nucleus, which could explain why nuclear permeabilization is required for reinitiation within one cell cycle. However, DeltaCDK mutants are retained in the nucleus, yet surprisingly they still support only one round of replication. As these highly conserved CDK sites are unnecessary for replication once per cell cycle, an alternative checkpoint role for monitoring completion of the S phase is suggested. PMID- 11018020 TI - A multifactor complex of eukaryotic initiation factors, eIF1, eIF2, eIF3, eIF5, and initiator tRNA(Met) is an important translation initiation intermediate in vivo. AB - Translation initiation factor 2 (eIF2) bound to GTP transfers the initiator methionyl tRNA to the 40S ribosomal subunit. The eIF5 stimulates GTP hydrolysis by the eIF2/GTP/Met-tRNA(i)(Met) ternary complex on base-pairing between Met tRNA(i)(Met) and the start codon. The eIF2, eIF5, and eIF1 all have been implicated in stringent selection of AUG as the start codon. The eIF3 binds to the 40S ribosome and promotes recruitment of the ternary complex; however, physical contact between eIF3 and eIF2 has not been observed. We show that yeast eIF5 can bridge interaction in vitro between eIF3 and eIF2 by binding simultaneously to the amino terminus of eIF3 subunit NIP1 and the amino-terminal half of eIF2beta, dependent on a conserved bipartite motif in the carboxyl terminus of eIF5. Additionally, the amino terminus of NIP1 can bind concurrently to eIF5 and eIF1. These findings suggest the occurrence of an eIF3/eIF1/eIF5/eIF2 multifactor complex, which was observed in cell extracts free of 40S ribosomes and found to contain stoichiometric amounts of tRNA(i)(Met). The multifactor complex was disrupted by the tif5-7A mutation in the bipartite motif of eIF5. Importantly, the tif5-7A mutant is temperature sensitive and displayed a substantial reduction in translation initiation at the restrictive temperature. We propose that the multifactor complex is an important intermediate in translation initiation in vivo. PMID- 11018021 TI - Uncoupling Raf1 from MEK1/2 impairs only a subset of cellular responses to Raf activation. AB - The Raf family of serine/threonine protein kinases is intimately involved in the transmission of cell regulatory signals controlling proliferation and differentiation. The best characterized Raf substrates are MEK1 and MEK2. The activation of MEK1/2 by Raf is required to mediate many of the cellular responses to Raf activation, suggesting that MEK1/2 are the dominant Raf effector proteins. However, accumulating evidence suggests that there are additional Raf substrates and that subsets of Raf-induced regulatory events are mediated independently of Raf activation of MEK1/2. To examine the possibility that there is bifurcation at the level of Raf in activation of MEK1/2-dependent and MEK1/2-independent cell regulatory events, we engineered a kinase-active Raf1 variant (RafBXB(T481A)) with an amino acid substitution that disrupts MEK1 binding. We find that disruption of MEK1/2 association uncouples Raf from activation of ERK1/2, induction of serum-response element-dependent gene expression, and induction of growth and morphological transformation. However, activation of NF-kappaB dependent gene expression and induction of neurite differentiation were unimpaired. In addition, Raf-dependent activation of p90 ribosomal S6 kinase was only slightly impaired. These results support the hypothesis that Raf kinases utilize multiple downstream effectors to regulate distinct cellular activities. PMID- 11018022 TI - Cloning and characterization of PHIP, a novel insulin receptor substrate-1 pleckstrin homology domain interacting protein. AB - Insulin receptor substrate-1 (IRS-1) protein is a major substrate of the insulin receptor tyrosine kinase and is essential for transducing many of the biological effects of insulin including mitogenesis, gene expression, and glucose transport. The N terminus of IRS-1 contains a pleckstrin homology (PH) domain that is critical for recognition and subsequent phosphorylation of IRS-1 by the activated insulin receptor. Here we report the isolation of a novel protein, PHIP (PH interacting protein), which selectively binds to the PH domain of IRS-1 in vitro and stably associates with IRS-1 in vivo. Importantly, mutants of the IRS-1 PH domain that disrupt the PH fold fail to bind to PHIP. Anti-phosphotyrosine immunoblots of PHIP revealed no discernible insulin receptor-regulated phosphorylation, suggesting that PHIP is not itself a substrate of the insulin receptor. In contrast to full-length PHIP, overexpression of the PH-binding region of PHIP has a pronounced inhibitory effect on insulin-induced IRS-1 tyrosine phosphorylation levels. Furthermore, expression of this dominant negative PHIP mutant leads to a marked attenuation of insulin-stimulated mitogen activated protein kinase activity. We conclude that PHIP represents a novel protein ligand of the IRS-1 PH domain that may serve to link IRS-1 to the insulin receptor. PMID- 11018023 TI - CLIF, a novel cycle-like factor, regulates the circadian oscillation of plasminogen activator inhibitor-1 gene expression. AB - The onset of myocardial infarction occurs frequently in the early morning, and it may partly result from circadian variation of fibrinolytic activity. Plasminogen activator inhibitor-1 activity shows a circadian oscillation and may account for the morning onset of myocardial infarction. However, the molecular mechanisms regulating this circadian oscillation remain unknown. Recent evidence indicates that basic helix-loop-helix (bHLH)/PAS domain transcription factors play a crucial role in controlling the biological clock that controls circadian rhythm. We isolated a novel bHLH/PAS protein, cycle-like factor (CLIF) from human umbilical vein endothelial cells. CLIF shares high homology with Drosophila CYCLE, one of the essential transcriptional regulators of circadian rhythm. CLIF is expressed in endothelial cells and neurons in the brain, including the suprachiasmatic nucleus, the center of the circadian clock. In endothelial cells, CLIF forms a heterodimer with CLOCK and up-regulates the PAI-1 gene through E-box sites. Furthermore, Period2 and Cryptochrome1, whose expression show a circadian oscillation in peripheral tissues, inhibit the PAI-1 promoter activation by the CLOCK:CLIF heterodimer. These results suggest that CLIF regulates the circadian oscillation of PAI-1 gene expression in endothelial cells. In addition, the results potentially provide a molecular basis for the morning onset of myocardial infarction. PMID- 11018024 TI - Structural requirements for the stabilization of metarhodopsin II by the C terminus of the alpha subunit of transducin. AB - The retinal receptor rhodopsin undergoes a conformational change upon light excitation to form metarhodopsin II (Meta II), which allows interaction and activation of its cognate G protein, transducin (G(t)). A C-terminal 11-amino acid peptide from transducin, G(talpha)-(340-350), has been shown to both bind and stabilize the Meta II conformation, mimicking heterotrimeric G(t). Using a combinatorial library we identified analogs of G(talpha)-(340-350) that bound light-activated rhodopsin with high affinity (Martin, E. L., Rens-Domiano, S., Schatz, P. J., and Hamm, H. E. (1996) J. Biol. Chem. 271, 361-366). We have made peptides with key substitutions either on the background of the native G(talpha) (340-350) sequence or on the high affinity sequences and used the stabilization of Meta II as a tool to determine which amino acids are critical in G protein rhodopsin interaction. Removal of the positive charge at the N termini by acylation or delocalization of the charge by K to R substitution enhances the affinity of the G(talpha)-(340-350) peptides for Meta II, whereas a decrease was observed following C-terminal amidation. Cys-347, a residue conserved in pertussis toxin-sensitive G proteins, was shown to interact with a hydrophobic site in Meta II. These studies provide further insight into the mechanism of interaction between the G(talpha) C terminus and light-activated rhodopsin. PMID- 11018025 TI - Ras-independent activation of the Raf/MEK/ERK pathway upon calcium-induced differentiation of keratinocytes. AB - MAPKs are crucially involved in the regulation of growth and differentiation of a variety of cells. To elucidate the role of MAPKs in keratinocyte differentiation, activation of ERK, JNK, and p38 in response to stimulation with extracellular calcium was analyzed. We provide evidence that calcium-induced differentiation of keratinocytes is associated with rapid and transient activation of the Raf/MEK/ERK pathway. Stimulation of keratinocytes with extracellular calcium resulted in activation of Raf isozymes and their downstream effector ERK within 10-15 min, but did not increase JNK or p38 activity. Calcium-induced ERK activation differed in kinetics from mitogenic ERK activation by epidermal growth factor and could be modulated by alterations of intracellular calcium levels. Interestingly, calcium stimulation led to down-regulation of Ras activity at the same time that ERK activation was initiated. Expression of a dominant-negative mutant of Ras also did not significantly impair calcium-induced ERK activation, indicating that calcium-mediated ERK activation does not require active Ras. Despite the transient nature of ERK activation, calcium-induced expression of the cyclin-dependent kinase inhibitor p21/Cip1 and the differentiation marker involucrin was sensitive to MEK inhibition, which suggests a role for the Raf/MEK/ERK pathway in early stages of keratinocyte differentiation. PMID- 11018026 TI - Purification and structural analysis of a soluble human chorionogonadotropin hormone-receptor complex. AB - Receptors for the luteotropin/human chorionogonadotropin hormone belong to the G protein-coupled receptor family by their membrane-anchoring domains. They also possess a large extracellular domain (ECD) responsible for most of the hormone receptor interactions. Structure-function studies identified several contacts between hormone and receptor ECD, but the precise topology of the complex is still unknown because of the lack of suitable heterologous expression means. Receptor ECDs exhibit leucine repeats and have been modelized on the basis of the three-dimensional structure of the porcine ribonuclease inhibitor, the first structurally known leucine-rich repeats protein. Here we report overexpression (up to 20 mg per liter) and purification to homogeneity of a soluble human chorionogonadotropin-ECD receptor complex secreted by stably cotransfected Chinese hamster ovary cells. Biochemical analysis and surface plasmon resonance data were in favor of a unique dimer with a 1:1 ligand-receptor stoichiometry. Immunopurified complex was submitted to circular dichroism characterization; CD spectra deconvolution indicated more than 25% alpha helices contributed by the receptor, in agreement with the porcine ribonuclease inhibitor-based modelization. PMID- 11018027 TI - Crystal structure of the CCAAT box/enhancer-binding protein beta activating transcription factor-4 basic leucine zipper heterodimer in the absence of DNA. AB - The crystal structure of the heterodimer formed by the basic leucine zipper (bZIP) domains of activating transcription factor-4 (ATF4) and CCAAT box/enhancer binding protein beta (C/EBP beta), from two different bZIP transcription factor families, has been determined and refined to 2.6 A. The structure shows that the heterodimer forms an asymmetric coiled-coil. Even in the absence of DNA, the basic region of ATF4 forms a continuous alpha-helix, but the basic region of C/EBP beta is disordered. Proteolysis, electrophoretic mobility shift assay, circular dichroism, and NMR analyses indicated that (i) the bZIP domain of ATF4 is a disordered monomer and forms a homodimer upon binding to the DNA target; (ii) the bZIP domain of ATF4 forms a heterodimer with the bZIP domain of C/EBP beta that binds the cAMP response element, but not CCAAT box DNA, with high affinity; and (iii) the basic region of ATF4 has a higher alpha-helical propensity than that of C/EBP beta. These results suggest that the degree of ordering of the basic region and the fork and the dimerization properties of the leucine zipper combine to distinguish the structurally similar bZIP domains of ATF4 and C/EBP beta with respect to DNA target sequence. This study provides insight into the mechanism by which dimeric bZIP transcription factors discriminate between closely related but distinct DNA targets. PMID- 11018028 TI - Sites of phosphorylation by protein kinase A in CDC25Mm/GRF1, a guanine nucleotide exchange factor for Ras. AB - Activation of the neuronal Ras GDP/GTP exchange factor (GEF) CDC25Mm/GRF1 is known to be associated with phosphorylation of serine/threonine. To increase our knowledge of the mechanism involved, we have analyzed the ability of several serine/threonine kinases to phosphorylate CDC25Mm in vivo and in vitro. We could demonstrate the involvement of cAMP-dependent protein kinase (PKA) in the phosphorylation of CDC25Mm in fibroblasts overexpressing this RasGEF as well as in mouse brain synaptosomal membranes. In vitro, PKA was found to phosphorylate multiple sites on purified CDC25Mm, in contrast to protein kinase C, calmodulin kinase II, and casein kinase II, which were virtually inactive. Eight phosphorylated serines and one threonine were identified by mass spectrometry and Edman degradation. Most of them were clustered around the Ras exchanger motif/PEST motifs situated in the C-terminal moiety (residues 631-978) preceding the catalytic domain. Ser745 and Ser822 were the most heavily phosphorylated residues and the only ones coinciding with PKA consensus sequences. Substitutions S745D and S822D showed that the latter mutation strongly inhibited the exchange activity of CDC25Mm on Ha-Ras. The multiple PKA-dependent phosphorylation sites on CDC25Mm suggest a complex regulatory picture of this RasGEF. The results are discussed in the light of structural and/or functional similarities with other members of this RasGEF family. PMID- 11018029 TI - Distinct oligomeric states of SMAD proteins in the transforming growth factor beta pathway. AB - Protein interactions are critical for the function of SMADs as mediators of transforming growth factor-beta (TGF-beta) signals. TGF-beta receptor phosphorylation of SMAD2 or SMAD3 causes their association with SMAD4 and accumulation in the nucleus where the SMAD complex binds cofactors that determine the choice of target genes. We provide evidence that in the basal state, SMADs 2, 3, and 4 form separate, strikingly different complexes. SMAD2 is found mostly as monomer, whereas the closely related SMAD3 exists in multiple oligomeric states. This difference is due to a unique structural element in the MH1 domain of SMAD2 that inhibits protein-protein interactions in the basal state. In contrast to SMAD2 and SMAD3, SMAD4 in the basal state is found mostly as a homo-oligomer, most likely a trimer. Upon cell stimulation with TGF-beta, SMAD proteins become engaged in a multitude of complexes ranging in size from SMAD2-SMAD4 heterodimers to assemblies of >650 kDa. The latter display the highest DNA binding affinity for the TGF-beta-response elements of JUNB and collagen 7. These observations, all validated with endogenous SMAD proteins, modify previous models regarding the assembly and activity of SMAD complexes in the TGF-beta pathway. PMID- 11018030 TI - YY1 as a regulator of replication-dependent hamster histone H3.2 promoter and an interactive partner of AP-2. AB - In analyzing cis-regulatory elements important for cell cycle control of the replication-dependent hamster histone H3.2 gene, we discovered a binding site for the transcription factor YY1 embedded within GC-rich sequences between the two tandem CCAAT repeats proximal to the TATA element. Base mutations that specifically eliminated YY1 binding resulted in suppression of the S phase induction of the H3.2 promoter. In addition, we discovered that YY1 is an interactive partner of AP-2, which also binds the H3.2 promoter and regulates its cell cycle-dependent expression. The critical domains for YY1 and AP-2A interaction are mapped, revealing that the N-terminal portion of YY1 (amino acids 1-300) and the DNA-binding/dimerization region of AP-2A are required. Our results suggest that YY1, acting as a transcription factor binding to its site on the promoter, or through protein-protein interaction with AP-2, may be part of a regulatory network including key cell cycle regulators such as c-Myc and Rb in controlling growth- and differentiation-regulated gene expression. PMID- 11018031 TI - Dimerization interfaces of v-erbA homodimers and heterodimers with retinoid X receptor alpha. AB - The oncoprotein v-ErbA, a member of the zinc finger transcription factor superfamily, is a mutated version of thyroid hormone receptor alpha1 that is virtually incapable of binding T3. v-ErbA and other members of this family can bind as homodimers and heterodimers with retinoid X receptors to specific DNA sequences arranged as direct, inverted, or everted repeats. At least two regions in the C-terminal domain, the I box (10 and 11 helices in v-ErbA and thyroid hormone receptors) and the 20-amino acid region are involved in dimerization. However, it has not been entirely understood how these receptors dimerize on differently oriented core motifs and whether the domain(s) responsible for homodimerization and heterodimerization are identical. Therefore, deletions of the entire 20-amino acid region, the 10 helix, the 11 helix, and point mutations within these regions of v-ErbA were made by site-directed mutagenesis. The mutant proteins were tested for their ability to form v-ErbA homodimers and heterodimers with retinoid X receptor alpha on differently oriented core motifs by electrophoretic mobility shift assay. Transient transfections were performed to determine the dominant negative activity of the v-ErbA mutants. The data indicate that different dimerization interfaces are used for v-ErbA homodimerization and heterodimerization with retinoid X receptor alpha, and different dimerization interfaces are used on differently oriented core motifs. The data are of general interest because the information improves our understanding of the role of these dimerization interfaces in the mechanism of action not only of v-ErbA but also of other members of the superfamily. PMID- 11018032 TI - Determinants of ligand binding specificity in the glial cell line-derived neurotrophic factor family receptor alpha S. AB - The glial cell line-derived neurotrophic factor (GDNF) family comprise a subclass of cystine-knot superfamily ligands that interact with a multisubunit receptor complex formed by the c-Ret tyrosine kinase and a cystine-rich glycosyl phosphatidylinositol-anchored binding subunit called GDNF family receptor alpha (GFRalpha). All four GDNF family ligands utilize c-Ret as a common signaling receptor, whereas specificity is conferred by differential binding to four distinct GFRalpha homologues. To understand how the different GFRalphas discriminate ligands, we have constructed a large set of chimeric and truncated receptors and analyzed their ligand binding and signaling capabilities. The major determinant of ligand binding was found in the most conserved region of the molecule, a central domain predicted to contain four conserved alpha helices and two beta strands. Distinct hydrophobic and positively charged residues in this central region were required for binding of GFRalpha1 to GDNF. Interaction of GFRalpha1 and GFRalpha2 with GDNF and neurturin required distinct subsegments within this central domain, which allowed the construction of chimeric receptors that responded equally well to both ligands. C-terminal segments adjacent to the central domain are necessary and have modulatory function in ligand binding. In contrast, the N-terminal domain was dispensable without compromising ligand binding specificity. Ligand-independent interaction with c-Ret also resides in the central domain of GFRalpha1, albeit within a distinct and smaller region than that required for ligand binding. Our results indicate that the central region of this class of receptors constitutes a novel binding domain for cystine-knot superfamily ligands. PMID- 11018033 TI - The minimal promoter of the human alpha 3 nicotinic receptor subunit gene. Molecular and functional characterization. AB - The minimal promoter of the human alpha(3) nicotinic receptor subunit gene has been mapped to a region of 60 base pairs and found to contain two Sp1 sites, one of which is essential for promoter activity. DNase footprinting has revealed the presence of another region of interaction with nuclear factors (named F2) immediately downstream of the Sp1 sites. This region has been found to be functional since it is capable of stimulating the minimal promoter. The F2 protection is completely and specifically competed by an AP2 consensus oligonucleotide that has been proved to bind AP2alpha exclusively. However, the AP2alpha recombinant protein was unable to bind the F2 region directly, thus suggesting that AP2alpha may participate in F2 protection by protein-protein interactions with other nuclear factors. The minimal promoter has been shown to be stimulated by two additional regions, one located downstream of F2 and the other upstream of the minimal promoter itself. In neuronal cells, the combined stimulatory activities of these three regions have synergistic effects, whereas in non-neuronal cells, there is a negative interference between the upstream and downstream regions. These opposite transcriptional effects may account for at least part of the neuro-specific expression profile of the alpha(3) gene. PMID- 11018034 TI - beta 2-adrenergic receptor-induced p38 MAPK activation is mediated by protein kinase A rather than by Gi or gbeta gamma in adult mouse cardiomyocytes. AB - Increasing evidence shows that stimulation of beta-adrenergic receptor (AR) activates mitogen-activated protein kinases (MAPKs), in addition to the classical G(s)-adenylyl cyclase-cAMP-dependent protein kinase (PKA) signaling cascade. In the present study, we demonstrate a novel beta(2)-AR-mediated cross-talk between PKA and p38 MAPK in adult mouse cardiac myocytes expressing beta(2)-AR, with a null background of beta(1)beta(2)-AR double knockout. beta(2)-AR stimulation by isoproterenol increased p38 MAPK activity in a time- and dose-dependent manner. Inhibiting G(i) with pertussis toxin or scavenging Gbetagamma with betaARK-ct overexpression could not prevent beta(2)-AR-induced p38 MAPK activation. In contrast, a specific peptide inhibitor of PKA, PKI (5 microm), completely abolished the stimulatory effect of beta(2)-AR, suggesting that beta(2)-AR induced p38 MAPK activation is mediated via a PKA-dependent mechanism, rather than by G(i) or Gbetagamma. This conclusion was further supported by the ability of forskolin (10 microm), an adenylyl cyclase activator, to elevate p38 MAPK activity in a PKI-sensitive manner. Furthermore, inhibition of p38 MAPK with SB203580 (10 microm) markedly enhanced the beta(2)-AR-mediated contractile response, without altering base-line contractility. These results provide the first evidence that cardiac beta(2)-AR activates p38 MAPK via a PKA-dependent signaling pathway, rather than by G(i) or Gbetagamma, and reveal a novel role of p38 MAPK in regulating cardiac contractility. PMID- 11018035 TI - Structure and orientation of two voltage-dependent anion-selective channel isoforms. An attenuated total reflection fourier-transform infrared spectroscopy study. AB - Two VDAC (voltage-dependent anion-selective channel) isoforms were purified from seed cotyledons of Phaseolus vulgaris by chromatofocusing chromatography. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was used to study the structural properties of the two isoforms reconstituted in a mixture of asolectin and 5% stigmasterol. The IR spectra of the two VDAC isoforms were highly similar indicating 50 to 53% anti-parallel beta-sheet. The orientation of the beta-strands relative to the barrel axis was calculated from the experimentally obtained dichroic ratios of the amide I beta-sheet component and the amide II band. Comparing the IR spectra of the reconstituted VDAC isoforms with the IR spectra of the bacterial porin OmpF, for which a high resolution structure is available, provided evidence for a general structural organization of the VDAC isoforms similar to that of bacterial porins. Hydrogen deuterium exchange measurements indicated that the exchange of the amide protons occurs to a higher extent in the two VDAC isoforms than in the OmpF porin. PMID- 11018036 TI - Suppression of preadipocyte differentiation and promotion of adipocyte death by HIV protease inhibitors. AB - Many human immunodeficiency virus (HIV)-infected patients taking combination antiretroviral therapy that includes HIV protease inhibitors experience atrophy of peripheral subcutaneous adipose tissue. We investigated the effects of HIV protease inhibitors on adipogenesis and adipocyte survival using the 3T3-L1 preadipocyte cell line. Several HIV protease inhibitors were found either to inhibit preadipocyte differentiation or to promote adipocyte cell death. One protease inhibitor, nelfinavir, elicited both of these effects strongly. When induced to differentiate in the presence of nelfinavir, 3T3-L1 preadipocytes failed to accumulate cytoplasmic triacylglycerol and failed to express normal levels of the adipogenic transcription factors CCAAT/enhancer-binding protein alpha and peroxisome proliferator-activated receptor gamma. The level of the proteolytically processed, active 68-kDa form of sterol regulatory element binding protein-1, a transcription factor known to promote lipogenic gene expression, also was reduced markedly in nelfinavir-treated cells, whereas the level of the 125-kDa precursor form of this protein was unaffected. The inhibitory effect of nelfinavir occurred subsequent to critical early events in preadipocyte differentiation, expression of CCAAT/enhancer-binding protein beta and completion of the mitotic clonal expansion phase, because these events were unaffected by nelfinavir treatment. In addition, nelfinavir treatment of fully differentiated 3T3-L1 adipocytes resulted in DNA strand cleavage and severe loss of cell viability. In contrast, cell proliferation and viability of preadipocytes were unaffected by nelfinavir treatment. Thus, molecular or cellular changes that occur during acquisition of the adipocyte phenotype promote susceptibility to nelfinavir-induced cell death. When considered together, these results suggest that nelfinavir may promote adipose tissue atrophy by compromising adipocyte viability and preventing replacement of lost adipocytes by inhibiting preadipocyte differentiation. PMID- 11018037 TI - Vascular endothelial growth factor induces expression of connective tissue growth factor via KDR, Flt1, and phosphatidylinositol 3-kinase-akt-dependent pathways in retinal vascular cells. AB - Fibroblastic proliferation accompanies many angiogenesis-related retinal and systemic diseases. Since connective tissue growth factor (CTGF) is a potent mitogen for fibrosis, extracellular matrix production, and angiogenesis, we have studied the effects and mechanism by which vascular endothelial growth factor (VEGF) regulates CTGF gene expression in retinal capillary cells. In our study, VEGF increased CTGF mRNA levels in a time- and concentration-dependent manner in bovine retinal endothelial cells and pericytes, without the need of new protein synthesis and without altering mRNA stability. VEGF activated the tyrosine receptor phosphorylation of KDR and Flt1 and increased the binding of phosphatidylinositol 3-kinase (PI3-kinase) p85 subunit to KDR and Flt1, both of which could mediate CTGF gene induction. VEGF-induced CTGF expression was mediated primarily by PI3-kinase activation, whereas PKC and ERK pathways made only minimal contributions. Furthermore, overexpression of constitutive active Akt was sufficient to induce CTGF gene expression, and inhibition of Akt activation by overexpressing dominant negative mutant of Akt abolished the VEGF induced CTGF expression. These data suggest that VEGF can increase CTGF gene expression in bovine retinal capillary cells via KDR or Flt receptors and the activation of PI3-kinase-Akt pathway independently of PKC or Ras-ERK pathway, possibly inducing the fibrosis observed in retinal neovascular diseases. PMID- 11018038 TI - Smad7-dependent regulation of heme oxygenase-1 by transforming growth factor-beta in human renal epithelial cells. AB - Heme oxygenase-1 (HO-1), a 32-kDa microsomal enzyme, is induced as a beneficial and adaptive response in cells/tissues exposed to oxidative stress. Transforming growth factor-beta1 (TGF-beta1) is a regulatory cytokine that has been implicated in a variety of renal diseases where it promotes extracellular matrix deposition and proinflammatory events. We hypothesize that the release of TGF-beta1 via autocrine and/or paracrine pathways may induce HO-1 and serve as a protective response in renal injury. To understand the molecular mechanism of HO-1 induction by TGF-beta1, we exposed confluent human renal proximal tubule cells to TGF-beta1 and observed a significant induction of HO-1 mRNA at 4 h with a maximal induction at 8 h. This induction was accompanied by increased expression of HO-1 protein. TGF-beta1 treatment in conjunction with actinomycin D or cycloheximide demonstrated that induction of HO-1 mRNA requires de novo transcription and, in part, protein synthesis. Exposure to TGF-beta1 resulted in marked induction of Smad7 mRNA with no effect on Smad6 expression. Overexpression of Smad7, but not Smad6, inhibited TGF-beta1-mediated induction of endogenous HO-1 gene expression. We speculate that the induction of HO-1 in the kidney is an adaptive response to the inflammatory effects of TGF-beta1 and manipulations of the Smad pathway to alter HO-1 expression may serve as a potential therapeutic target. PMID- 11018039 TI - G protein beta gamma subunits inhibit nongenomic progesterone-induced signaling and maturation in Xenopus laevis oocytes. Evidence for a release of inhibition mechanism for cell cycle progression. AB - Progesterone-induced maturation of Xenopus oocytes is a well known example of nongenomic signaling by steroids; however, little is known about the early signaling events involved in this process. Previous work has suggested that G proteins and G protein-coupled receptors may be involved in progesterone-mediated oocyte maturation as well as in other nongenomic steroid-induced signaling events. To investigate the role of G proteins in nongenomic signaling by progesterone, the effects of modulating Galpha and Gbetagamma levels in Xenopus oocytes on progesterone-induced signaling and maturation were examined. Our results demonstrate that Gbetagamma subunits, rather than Galpha, are the principal mediators of progesterone action in this system. We show that overexpression of Gbetagamma inhibits both progesterone-induced maturation and activation of the MAPK pathway, whereas sequestration of endogenous Gbetagamma subunits enhances progesterone-mediated signaling and maturation. These data are consistent with a model whereby endogenous free Xenopus Gbetagamma subunits constitutively inhibit oocyte maturation. Progesterone may induce maturation by antagonizing this inhibition and therefore allowing cell cycle progression to occur. These studies offer new insight into the early signaling events mediated by progesterone and may be useful in characterizing and identifying the membrane progesterone receptor in oocytes. PMID- 11018040 TI - Identification and characterization of two novel components of the Prp19p associated complex, Ntc30p and Ntc20p. AB - The yeast Saccharomyces cerevisiae Prp19p protein is an essential splicing factor and a spliceosomal component. It is not tightly associated with small nuclear RNAs (snRNAs) but is associated with a protein complex consisting of at least eight proteins. We have identified two novel components of the Prp19p-associated complex, Ntc30p and Ntc20p. Like other identified components of the complex, both Ntc30p and Ntc20p are associated with the spliceosome in the same manner as Prp19p immediately after or concurrently with dissociation of U4, indicating that the entire complex may bind to the spliceosome as an intact form. Neither Ntc30p nor Ntc20p directly interacts with Prp19p, but both interact with another component of the complex, Ntc85p. Immunoprecipitation analysis revealed an ordered interactions of these components in formation of the Prp19p-associated complex. Although null mutants of NTC30 or NTC20 showed no obvious growth phenotype, deletion of both genes impaired yeast growth resulting in accumulation of precursor mRNA. Extracts prepared from such a strain were defective in pre mRNA splicing in vitro, but the splicing activity could be restored upon addition of the purified Prp19p-associated complex. These results indicate that Ntc30p and Ntc20p are auxiliary splicing factors the functions of which may be modulating the function of the Prp19p-associated complex. PMID- 11018041 TI - Oligomeric forms of the metastasis-related Mts1 (S100A4) protein stimulate neuronal differentiation in cultures of rat hippocampal neurons. AB - Neuronal differentiation and axonal growth are controlled by a variety of factors including neurotrophic factors, extracellular matrix components, and cell adhesion molecules. Here we describe a novel and very efficient neuritogenic factor, the metastasis-related Mts1 protein, belonging to the S100 protein family. The oligomeric but not the dimeric form of Mts1 strongly induces differentiation of cultured hippocampal neurons. A mutant with a single Y75F amino acid substitution, which stabilizes the dimeric form of Mts1, is unable to promote neurite extension. Disulfide bonds do not play an essential role in the Mts1 neuritogenic activity. Mts1-stimulated neurite outgrowth involves activation of phospholipase C and protein kinase C, depends on the intracellular level of Ca(2+), and requires activation of the extracellular signal-regulated kinases (ERKs) 1 and 2. PMID- 11018042 TI - Specific activation of LIM kinase 2 via phosphorylation of threonine 505 by ROCK, a Rho-dependent protein kinase. AB - LIM-kinase 1 (LIMK1) and LIM-kinase 2 (LIMK2) regulate actin cytoskeletal reorganization via cofilin phosphorylation downstream of distinct Rho family GTPases. We report our findings that ROCK, a downstream protein kinase of Rho, specifically activates LIMK2 but not LIMK1 downstream of RhoA. LIMK1 and LIMK2 activities toward cofilin phosphorylation were stimulated by co-expression with the active form of ROCK (ROCK-Delta3), whereas full-length ROCK selectively activates LIMK2 but not LIMK1. Activation of LIMK2 by RhoA was inhibited by Y 27632, a specific inhibitor of ROCK, but Rac1-mediated activation of LIMK1 was not. ROCK directly phosphorylated the threonine 505 residue within the activation segment of LIMK2 and markedly stimulated LIMK2 activity. A LIMK2 mutant with replacement of threonine 505 by valine abolished LIMK2 activities for cofilin phosphorylation and actin cytoskeletal changes, whereas replacement by glutamate enhanced the protein kinase activity and stress fiber formation by LIMK2. These results indicate that ROCK directly phosphorylates threonine 505 and activates LIMK2 downstream of RhoA and that this phosphorylation is essential for LIMK2 to induce actin cytoskeletal reorganization. Together with the finding that LIMK1 is regulated by Pak1, LIMK1 and LIMK2 are regulated by different protein kinases downstream of distinct Rho family GTPases. PMID- 11018043 TI - Disulfide bonds of GM2 synthase homodimers. Antiparallel orientation of the catalytic domains. AB - GM2 synthase is a homodimer in which the subunits are joined by lumenal domain disulfide bond(s). To define the disulfide bond pattern of this enzyme, we analyzed a soluble form by chemical fragmentation, enzymatic digestion, and mass spectrometry and a full-length form by site-directed mutagenesis. All Cys residues of the lumenal domain of GM2 synthase are disulfide bonded with Cys(429) and Cys(476) forming a disulfide-bonded pair while Cys(80) and Cys(82) are disulfide bonded in combination with Cys(412) and Cys(529). Partial reduction to produce monomers converted Cys(80) and Cys(82) to free thiols while the Cys(429) to Cys(476) disulfide remained intact. CNBr cleavage at amino acid 330 produced a monomer-sized band under nonreducing conditions which was converted upon reduction to a 40-kDa fragment and a 24-kDa myc-positive fragment. Double mutation of Cys(80) and Cys(82) to Ser produced monomers but not dimers. In summary these results demonstrate that Cys(429) and Cys(476) form an intrasubunit disulfide while the intersubunit disulfides formed by both Cys(80) and Cys(82) with Cys(412) and Cys(529) are responsible for formation of the homodimer. This disulfide bond arrangement results in an antiparallel orientation of the catalytic domains of the GM2 synthase homodimer. PMID- 11018044 TI - SOCS3 mediates feedback inhibition of the leptin receptor via Tyr985. AB - During leptin signaling, each of the phosphorylated tyrosine residues on the long form of the leptin receptor (LRb) mediates distinct signals. Phosphorylated Tyr(1138) binds STAT3 to mediate its tyrosine phosphorylation and transcriptional activation, while phosphorylated Tyr(985) binds the tyrosine phosphatase SHP-2 and reportedly mediates both activation of ERK kinases and inhibition of LRb mediated STAT3 activation. We show here that although mutation of Tyr(985) does not alter STAT3 signaling by erythropoietin receptor-LRb (ELR) chimeras in transfected 293 cells at short times of stimulation, this mutation enhances STAT3 signaling at longer times of stimulation (>6 h). These data suggest that Tyr(985) may mediate feedback inhibition of LRb signaling by an LRb-induced LRb inhibitor, such as SOCS3. Indeed, SOCS3 binds specifically to phosphorylated Tyr(985) of LRb, and SOCS3 fails to inhibit transcription by ELR following mutation of Tyr(985), suggesting that SOCS3 inhibits LRb signaling by binding to phosphorylated Tyr(985). Additionally, overexpression of SOCS3, but not SHP-2, impairs ELR signaling, and the overexpression of SHP-2 blunts SOCS3-mediated inhibition of ELR signaling. Thus, our data suggest that in addition to mediating SHP-2 binding and ERK activation during acute stimulation, Tyr(985) of LRb mediates feedback inhibition of LRb signaling by binding to LRb-induced SOCS3. PMID- 11018045 TI - Mechanism of Cu,Zn-superoxide dismutase activation by the human metallochaperone hCCS. AB - The mechanism for copper loading of the antioxidant enzyme copper, zinc superoxide dismutase (SOD1) by its partner metallochaperone protein is not well understood. Here we show the human copper chaperone for Cu,Zn-SOD1 (hCCS) activates either human or yeast enzymes in vitro by direct protein to protein transfer of the copper cofactor. Interestingly, when denatured with organic solvents, the apo-form of human SOD1 cannot be reactivated by added copper ion alone, suggesting an additional function of hCCS such as facilitation of an active folded state of the enzyme. While hCCS can bind several copper ions, metal binding studies in the presence of excess copper scavengers that mimic the intracellular chelation capacity indicate a limiting stoichiometry of one copper and one zinc per hCCS monomer. This protein is active and unlike the yeast protein, is a homodimer regardless of copper occupancy. Matrix-assisted laser desorption ionization-mass spectrometry and metal binding studies suggest that Cu(I) is bound by residues from the first and third domains and no bound copper is detected for the second domain of hCCS in either the full-length or truncated forms of the protein. Copper-induced conformational changes in the essential C terminal peptide of hCCS are consistent with a "pivot, insert, and release" mechanism that is similar to one proposed for the well characterized metal handling enzyme, mercuric ion reductase. PMID- 11018046 TI - Mapping the pathways for O2 entry into and exit from myoglobin. AB - The effects of mutagenesis on geminate and bimolecular O2 rebinding to 90 mutants at 27 different positions were used to map pathways for ligand movement into and out of sperm whale myoglobin. By analogy to a baseball glove, the protein "catches" and then "holds" incoming ligand molecules long enough to allow bond formation with the iron atom. Opening of the glove occurs by outward movements of the distal histidine (His(64)), and the ligands are trapped in the interior "webbing" of the distal pocket, in the space surrounded by Ile(28), Leu(29), Leu(32), Val(68), and Ile(107). The size of this pocket is a major determinant of the rate of ligand entry into the protein. Immediately after photo- or thermal dissociation, O2 moves away from the iron into this interior pocket. The majority of the dissociated ligands return to the active site and either rebind to the iron atom or escape through the His(64) gate. A fraction of the ligands migrate further away from the heme group into cavities that have been defined as Xe binding sites 4 and 1; however, most of these ligands also return to the distal pocket, and net escape through the interior of wild-type myoglobin is <20-25%. PMID- 11018047 TI - Regulation of asymmetric smooth muscle myosin II molecules. AB - The emerging view of smooth/nonmuscle myosin regulation suggests that the attainment of the completely inhibited state requires numerous weak interactions between components of the two heads and the myosin rod. To further examine the nature of the structural requirements for regulation, we engineered smooth muscle heavy meromyosin molecules that contained one complete head and truncations of the second head. These truncations eliminated the motor domain but retained two, one, or no light chains. All constructs contained 37 heptads of rod sequence. None of the truncated constructs displayed complete regulation of both ATPase and motility, reinforcing the idea that interactions between motor domains are necessary for complete regulation. Surprisingly, the rate of ADP release was slowed by regulatory light chain dephosphorylation of the truncated construct that contained all four light chains and one motor domain. These data suggest that there is a second step (ADP release) in the smooth muscle myosin-actin activated ATPase cycle that is modulated by regulatory light chain phosphorylation. This may be part of the mechanism underlying "latch" in smooth muscle. PMID- 11018048 TI - Structural conservation of neurotropism-associated VspA within the variable Borrelia Vsp-OspC lipoprotein family. AB - Vsp surface lipoproteins are serotype-defining antigens of relapsing fever spirochetes that undergo multiphasic antigenic variation to avoid the immune response. One of these proteins, VspA of Borrelia turicatae, is also associated with neurotropism in infected mice. Vsp proteins are highly polymorphic in sequence, which may relate to their specific antibody reactivities and host cell interactions. To determine whether sequence variations affect protein structure, we compared B. turicatae VspA with three related proteins: VspB of B. turicatae, Vsp26 of the relapsing fever agent Borrelia hermsii, and OspC of the Lyme disease spirochete Borrelia burgdorferi. Recombinant non-lipidated proteins were purified by affinity or ion exchange chromatography. Circular dichroism spectra revealed similar, highly alpha-helical secondary structures for all four proteins. In vitro assays demonstrated protease-resistant, thermostable Vsp cores starting at a conserved serine at position 34 (Ser(34)). All proteins aggregate as dimers in solution. In situ trypsin treatment and surface protein cross-linking showed that the native lipoproteins also form protease-resistant dimers. These findings indicate that Vsp proteins have a common compact fold and that their established functions are based on localized polymorphisms. Two forms of VspA crystals suitable for structure determination by x-ray diffraction methods have been obtained. PMID- 11018049 TI - Protein ligands to HuR modulate its interaction with target mRNAs in vivo. AB - AU-rich elements (AREs) present in the 3' untranslated regions of many protooncogene, cytokine, and lymphokine messages target them for rapid degradation. HuR, a ubiquitously expressed member of the ELAV (embryonic lethal abnormal vision) family of RNA binding proteins, selectively binds AREs and stabilizes ARE-containing mRNAs in transiently transfected cells. Here, we identify four mammalian proteins that bind regions of HuR known to be essential for its ability to shuttle between the nucleus and the cytoplasm and to stabilize mRNA: SETalpha, SETbeta, pp32, and acidic protein rich in leucine (APRIL). Three have been reported to be protein phosphatase 2A inhibitors. All four ligands contain long, acidic COOH-terminal tails, while pp32 and APRIL share a second motif: rev-like leucine-rich repeats in their NH(2)-terminal regions. We show that pp32 and APRIL are nucleocytoplasmic shuttling proteins that interact with the nuclear export factor CRM1 (chromosomal region maintenance protein 1). The inhibition of CRM1 by leptomycin B leads to the nuclear retention of pp32 and APRIL, their increased association with HuR, and an increase in HuR's association with nuclear poly(A)+ RNA. Furthermore, transcripts from the ARE-containing c-fos gene are selectively retained in the nucleus, while the cytoplasmic distribution of total poly(A)+ RNA is not altered. These data provide evidence that interaction of its ligands with HuR modulate HuR's ability to bind its target mRNAs in vivo and suggest that CRM1 is instrumental in the export of at least some cellular mRNAs under certain conditions. We discuss the possible role of these ligands upstream of HuR in pathways that govern the stability of ARE containing mRNAs. PMID- 11018050 TI - Tea2p is a kinesin-like protein required to generate polarized growth in fission yeast. AB - Cytoplasmic microtubules are critical for establishing and maintaining cell shape and polarity. Our investigations of kinesin-like proteins (klps) and morphological mutants in the fission yeast Schizosaccharomyces pombe have identified a kinesin-like gene, tea2(+), that is required for cells to generate proper polarized growth. Cells deleted for this gene are often bent during exponential growth and initiate growth from improper sites as they exit stationary phase. They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells. The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p. Tea2p is localized to the tips of the cell and in a punctate pattern within the cell, often coincident with the ends of cytoplasmic microtubules. These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell. PMID- 11018051 TI - Cortactin localization to sites of actin assembly in lamellipodia requires interactions with F-actin and the Arp2/3 complex. AB - Cortactin is an actin-binding protein that is enriched within the lamellipodia of motile cells and in neuronal growth cones. Here, we report that cortactin is localized with the actin-related protein (Arp) 2/3 complex at sites of actin polymerization within the lamellipodia. Two distinct sequence motifs of cortactin contribute to its interaction with the cortical actin network: the fourth of six tandem repeats and the amino-terminal acidic region (NTA). Cortactin variants lacking either the fourth tandem repeat or the NTA failed to localize at the cell periphery. Tandem repeat four was necessary for cortactin to stably bind F-actin in vitro. The NTA region interacts directly with the Arp2/3 complex based on affinity chromatography, immunoprecipitation assays, and binding assays using purified components. Cortactin variants containing the NTA region were inefficient at promoting Arp2/3 actin nucleation activity. These data provide strong evidence that cortactin is specifically localized to sites of dynamic cortical actin assembly via simultaneous interaction with F-actin and the Arp2/3 complex. Cortactin interacts via its Src homology 3 (SH3) domain with ZO-1 and the SHANK family of postsynaptic density 95/dlg/ZO-1 homology (PDZ) domain containing proteins, suggesting that cortactin contributes to the spatial organization of sites of actin polymerization coupled to selected cell surface transmembrane receptor complexes. PMID- 11018052 TI - Agrin isoforms with distinct amino termini: differential expression, localization, and function. AB - The proteoglycan agrin is required for postsynaptic differentiation at the skeletal neuromuscular junction, but is also associated with basal laminae in numerous other tissues, and with the surfaces of some neurons. Little is known about its roles at sites other than the neuromuscular junction, or about how its expression and subcellular localization are regulated in any tissue. Here we demonstrate that the murine agrin gene generates two proteins with different NH(2) termini, and present evidence that these isoforms differ in subcellular localization, tissue distribution, and function. The two isoforms share approximately 1,900 amino acids (aa) of common sequence following unique NH(2) termini of 49 or 150 aa; we therefore call them short NH(2)-terminal (SN) and long NH(2)-terminal (LN) isoforms. In the mouse genome, LN-specific exons are upstream of an SN-specific exon, which is in turn upstream of common exons. LN agrin is expressed in both neural and nonneural tissues. In spinal cord it is expressed in discrete subsets of cells, including motoneurons. In contrast, SN agrin is selectively expressed in the nervous system but is widely distributed in many neuronal cell types. Both isoforms are externalized from cells but LN-agrin assembles into basal laminae whereas SN-agrin remains cell associated. Differential expression of the two isoforms appears to be transcriptionally regulated, whereas the unique SN and LN sequences direct their distinct subcellular localizations. Insertion of a "gene trap" construct into the mouse genome between the LN and SN exons abolished expression of LN-agrin with no detectable effect on expression levels of SN-agrin or on SN-agrin bioactivity in vitro. Agrin protein was absent from all basal laminae in mice lacking LN-agrin transcripts. The formation of the neuromuscular junctions was as drastically impaired in these mutants as in mice lacking all forms of agrin. Thus, basal lamina-associated LN-agrin is required for neuromuscular synaptogenesis, whereas cell-associated SN-agrin may play distinct roles in the central nervous system. PMID- 11018053 TI - Synbindin, A novel syndecan-2-binding protein in neuronal dendritic spines. AB - Dendritic spines are small protrusions on the surface of dendrites that receive the vast majority of excitatory synapses. We previously showed that the cell surface heparan sulfate proteoglycan syndecan-2 induces spine formation upon transfection into hippocampal neurons. This effect requires the COOH-terminal EFYA sequence of syndecan-2, suggesting that cytoplasmic molecules interacting with this sequence play a critical role in spine morphogenesis. Here, we report a novel protein that binds to the EFYA motif of syndecan-2. This protein, named synbindin, is expressed by neurons in a pattern similar to that of syndecan-2, and colocalizes with syndecan-2 in the spines of cultured hippocampal neurons. In transfected hippocampal neurons, synbindin undergoes syndecan-2-dependent clustering. Synbindin is structurally related to yeast proteins known to be involved in vesicle transport. Immunoelectron microscopy localized synbindin on postsynaptic membranes and intracellular vesicles within dendrites, suggesting a role in postsynaptic membrane trafficking. Synbindin coimmunoprecipitates with syndecan-2 from synaptic membrane fractions. Our results show that synbindin is a physiological syndecan-2 ligand on dendritic spines. We suggest that syndecan-2 induces spine formation by recruiting intracellular vesicles toward postsynaptic sites through the interaction with synbindin. PMID- 11018054 TI - Endoplasmic reticulum degradation requires lumen to cytosol signaling. Transmembrane control of Hrd1p by Hrd3p. AB - Endoplasmic reticulum (ER)-associated degradation (ERAD) is required for ubiquitin-mediated destruction of numerous proteins. ERAD occurs by processes on both sides of the ER membrane, including lumenal substrate scanning and cytosolic destruction by the proteasome. The ER resident membrane proteins Hrd1p and Hrd3p play central roles in ERAD. We show that these two proteins directly interact through the Hrd1p transmembrane domain, allowing Hrd1p stability by Hrd3p dependent control of the Hrd1p RING-H2 domain activity. Rigorous reevaluation of Hrd1p topology demonstrated that the Hrd1p RING-H2 domain is located and functions in the cytosol. An engineered, completely lumenal, truncated version of Hrd3p functioned normally in both ERAD and Hrd1p stabilization, indicating that the lumenal domain of Hrd3p regulates the cytosolic Hrd1p RING-H2 domain by signaling through the Hrd1p transmembrane domain. Additionally, we identified a lumenal region of Hrd3p dispensable for regulation of Hrd1p stability, but absolutely required for normal ERAD. Our studies show that Hrd1p and Hrd3p form a stoichiometric complex with ERAD determinants in both the lumen and the cytosol. The HRD complex engages in lumen to cytosol communication required for regulation of Hrd1p stability and the coordination of ERAD events on both sides of the ER membrane. PMID- 11018055 TI - Dishevelled-1 regulates microtubule stability: a new function mediated by glycogen synthase kinase-3beta. AB - Dishevelled has been implicated in the regulation of cell fate decisions, cell polarity, and neuronal function. However, the mechanism of Dishevelled action remains poorly understood. Here we examine the cellular localization and function of the mouse Dishevelled protein, DVL-1. Endogenous DVL-1 colocalizes with axonal microtubules and sediments with brain microtubules. Expression of DVL-1 protects stable microtubules from depolymerization by nocodazole in both dividing cells and differentiated neuroblastoma cells. Deletion analyses reveal that the PDZ domain, but not the DEP domain, of DVL-1 is required for microtubule stabilization. The microtubule stabilizing function of DVL-1 is mimicked by lithium-mediated inhibition of glycogen synthase kinase-3beta (GSK-3beta) and blocked by expression of GSK-3beta. These findings suggest that DVL-1, through GSK-3beta, can regulate microtubule dynamics. This new function of DVL-1 in controlling microtubule stability may have important implications for Dishevelled proteins in regulating cell polarity. PMID- 11018056 TI - Meiotic telomere protein Ndj1p is required for meiosis-specific telomere distribution, bouquet formation and efficient homologue pairing. AB - We have investigated the requirements for NDJ1 in meiotic telomere redistribution and clustering in synchronized cultures of Saccharomyces cerevisiae. On induction of wild-type meiosis, telomeres disperse from premeiotic aggregates over the nuclear periphery, and then cluster near the spindle pole body (bouquet arrangement) before dispersing again. In ndj1Delta meiocytes, telomeres are scattered throughout the nucleus and fail to form perinuclear meiosis-specific distribution patterns, suggesting that Ndj1p may function to tether meiotic telomeres to the nuclear periphery. Since ndj1Delta meiocytes fail to cluster their telomeres at any prophase stage, Ndj1p is the first protein shown to be required for bouquet formation in a synaptic organism. Analysis of homologue pairing by two-color fluorescence in situ hybridization with cosmid probes to regions on III, IX, and XI revealed that disruption of bouquet formation is associated with a significant delay (>2 h) of homologue pairing. An increased and persistent fraction of ndj1Delta meiocytes with Zip1p polycomplexes suggests that chromosome polarization is important for synapsis progression. Thus, our observations support the hypothesis that meiotic telomere clustering contributes to efficient homologue alignment and synaptic pairing. Under naturally occurring conditions, bouquet formation may allow for rapid sporulation and confer a selective advantage. PMID- 11018057 TI - Rapid transport of internalized P-selectin to late endosomes and the TGN: roles in regulating cell surface expression and recycling to secretory granules. AB - Prior studies on receptor recycling through late endosomes and the TGN have suggested that such traffic may be largely limited to specialized proteins that reside in these organelles. We present evidence that efficient recycling along this pathway is functionally important for nonresident proteins. P-selectin, a transmembrane cell adhesion protein involved in inflammation, is sorted from recycling cell surface receptors (e.g., low density lipoprotein [LDL] receptor) in endosomes, and is transported from the cell surface to the TGN with a half time of 20-25 min, six to seven times faster than LDL receptor. Native P-selectin colocalizes with LDL, which is efficiently transported to lysosomes, for 20 min after internalization, but a deletion mutant deficient in endosomal sorting activity rapidly separates from the LDL pathway. Thus, P-selectin is sorted from LDL receptor in early endosomes, driving P-selectin rapidly into late endosomes. P-selectin then recycles to the TGN as efficiently as other receptors. Thus, the primary effect of early endosomal sorting of P-selectin is its rapid delivery to the TGN, with rapid turnover in lysosomes a secondary effect of frequent passage through late endosomes. This endosomal sorting event provides a mechanism for efficiently recycling secretory granule membrane proteins and, more generally, for downregulating cell surface receptors. PMID- 11018059 TI - Functional redundancy in the maize meiotic kinetochore. AB - Kinetochores can be thought of as having three major functions in chromosome segregation: (a) moving plateward at prometaphase; (b) participating in spindle checkpoint control; and (c) moving poleward at anaphase. Normally, kinetochores cooperate with opposed sister kinetochores (mitosis, meiosis II) or paired homologous kinetochores (meiosis I) to carry out these functions. Here we exploit three- and four-dimensional light microscopy and the maize meiotic mutant absence of first division 1 (afd1) to investigate the properties of single kinetochores. As an outcome of premature sister kinetochore separation in afd1 meiocytes, all of the chromosomes at meiosis II carry single kinetochores. Approximately 60% of the single kinetochore chromosomes align at the spindle equator during prometaphase/metaphase II, whereas acentric fragments, also generated by afd1, fail to align at the equator. Immunocytochemistry suggests that the plateward movement occurs in part because the single kinetochores separate into half kinetochore units. Single kinetochores stain positive for spindle checkpoint proteins during prometaphase, but lose their staining as tension is applied to the half kinetochores. At anaphase, approximately 6% of the kinetochores develop stable interactions with microtubules (kinetochore fibers) from both spindle poles. Our data indicate that maize meiotic kinetochores are plastic, redundant structures that can carry out each of their major functions in duplicate. PMID- 11018058 TI - Glycogen synthase kinase-3beta is a negative regulator of cardiomyocyte hypertrophy. AB - Hypertrophy is a basic cellular response to a variety of stressors and growth factors, and has been best characterized in myocytes. Pathologic hypertrophy of cardiac myocytes leads to heart failure, a major cause of death and disability in the developed world. Several cytosolic signaling pathways have been identified that transduce prohypertrophic signals, but to date, little work has focused on signaling pathways that might negatively regulate hypertrophy. Herein, we report that glycogen synthase kinase-3beta (GSK-3beta), a protein kinase previously implicated in processes as diverse as development and tumorigenesis, is inactivated by hypertrophic stimuli via a phosphoinositide 3-kinase-dependent protein kinase that phosphorylates GSK-3beta on ser 9. Using adenovirus-mediated gene transfer of GSK-3beta containing a ser 9 to alanine mutation, which prevents inactivation by hypertrophic stimuli, we demonstrate that inactivation of GSK 3beta is required for cardiomyocytes to undergo hypertrophy. Furthermore, our data suggest that GSK-3beta regulates the hypertrophic response, at least in part, by modulating the nuclear/cytoplasmic partitioning of a member of the nuclear factor of activated T cells family of transcription factors. The identification of GSK-3beta as a transducer of antihypertrophic signals suggests that novel therapeutic strategies to treat hypertrophic diseases of the heart could be designed that target components of the GSK-3 pathway. PMID- 11018060 TI - Assembly of myelin by association of proteolipid protein with cholesterol- and galactosylceramide-rich membrane domains. AB - Myelin is a specialized membrane enriched in glycosphingolipids and cholesterol that contains a limited spectrum of proteins. We investigated the assembly of myelin components by oligodendrocytes and analyzed the role of lipid-protein interactions in this process. Proteolipid protein (PLP), the major myelin protein, was recovered from cultured oligodendrocytes from a low-density CHAPS insoluble membrane fraction (CIMF) enriched in myelin lipids. PLP associated with the CIMF after leaving the endoplasmic reticulum but before exiting the Golgi apparatus, suggesting that myelin lipid and protein components assemble in the Golgi complex. The specific association of PLP with myelin lipids in CIMF was supported by the finding that it was efficiently cross-linked to photoactivable cholesterol, but not to phosphatidylcholine, which is underrepresented in both myelin and CIMF. Furthermore, depletion of cholesterol or inhibition of sphingolipid synthesis in oligodendrocytes abolished the association of PLP with CIMF. Thus, PLP may be recruited to myelin rafts, represented by CIMF, via lipid protein interactions. In contrast to oligodendrocytes, after transfection in BHK cells, PLP is absent from isolated CIMF, suggesting that PLP requires specific lipids for raft association. In mice deficient in the enzyme ceramide galactosyl transferase, which cannot synthesize the main myelin glycosphingolipids, a large fraction of PLP no longer associates with rafts. Formation of a cholesterol- and galactosylceramide-rich membrane domain (myelin rafts) may be critical for the sorting of PLP and assembly of myelin in oligodendrocytes. PMID- 11018061 TI - Dynein, dynactin, and kinesin II's interaction with microtubules is regulated during bidirectional organelle transport. AB - The microtubule motors, cytoplasmic dynein and kinesin II, drive pigmented organelles in opposite directions in Xenopus melanophores, but the mechanism by which these or other motors are regulated to control the direction of organelle transport has not been previously elucidated. We find that cytoplasmic dynein, dynactin, and kinesin II remain on pigment granules during aggregation and dispersion in melanophores, indicating that control of direction is not mediated by a cyclic association of motors with these organelles. However, the ability of dynein, dynactin, and kinesin II to bind to microtubules varies as a function of the state of aggregation or dispersion of the pigment in the cells from which these molecules are isolated. Dynein and dynactin bind to microtubules when obtained from cells with aggregated pigment, whereas kinesin II binds to microtubules when obtained from cells with dispersed pigment. Moreover, the microtubule binding activity of these motors/dynactin can be reversed in vitro by the kinases and phosphatase that regulate the direction of pigment granule transport in vivo. These findings suggest that phosphorylation controls the direction of pigment granule transport by altering the ability of dynein, dynactin, and kinesin II to interact with microtubules. PMID- 11018062 TI - Spontaneous release of cytosolic proteins from posttranslational substrates before their transport into the endoplasmic reticulum. AB - In posttranslational translocation in yeast, completed protein substrates are transported across the endoplasmic reticulum membrane through a translocation channel formed by the Sec complex. We have used photo-cross-linking to investigate interactions of cytosolic proteins with a substrate synthesized in a reticulocyte lysate system, before its posttranslational translocation through the channel in the yeast membrane. Upon termination of translation, the signal recognition particle (SRP) and the nascent polypeptide-associated complex (NAC) are released from the polypeptide chain, and the full-length substrate interacts with several different cytosolic proteins. At least two distinct complexes exist that contain among other proteins either 70-kD heat shock protein (Hsp70) or tailless complex polypeptide 1 (TCP1) ring complex/chaperonin containing TCP1 (TRiC/CCT), which keep the substrate competent for translocation. None of the cytosolic factors appear to interact specifically with the signal sequence. Dissociation of the cytosolic proteins from the substrate is accelerated to the same extent by the Sec complex and an unspecific GroEL trap, indicating that release occurs spontaneously without the Sec complex playing an active role. Once bound to the Sec complex, the substrate is stripped of all cytosolic proteins, allowing it to subsequently be transported through the membrane channel without the interference of cytosolic binding partners. PMID- 11018063 TI - Microtubule and motor-dependent endocytic vesicle sorting in vitro. AB - Endocytic vesicles undergo fission to sort ligand from receptor. Using quantitative immunofluorescence and video imaging, we provide the first in vitro reconstitution of receptor-ligand sorting in early endocytic vesicles derived from rat liver. We show that to undergo fission, presegregation vesicles must bind to microtubules (MTs) and move upon addition of ATP. Over 13% of motile vesicles elongate and are capable of fission. After fission, one vesicle continues to move, whereas the other remains stationary, resulting in their separation. On average, almost 90% receptor is found in one daughter vesicle, whereas ligand is enriched by approximately 300% with respect to receptor in the other daughter vesicle. Although studies performed on polarity marked MTs showed approximately equal plus and minus end-directed motility, immunofluorescence microscopy revealed that kinesins, but not dynein, were associated with these vesicles. Motility and fission were prevented by addition of 1 mM 5' adenylylimido-diphosphate (AMP-PNP, an inhibitor of kinesins) or incubation with kinesin antibodies, but were unaffected by addition of 5 microM vanadate (a dynein inhibitor) or dynein antibodies. These studies indicate an essential role of kinesin-based MT motility in endocytic vesicle sorting, providing a system in which factors required for endocytic vesicle processing can be identified and characterized. PMID- 11018064 TI - Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. AB - The dynamin family of large GTPases has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. It is believed that dynamin interacts with a variety of cellular proteins to constrict membranes. The actin cytoskeleton has also been implicated in altering membrane shape and form during cell migration, endocytosis, and secretion and has been postulated to work synergistically with dynamin and coat proteins in several of these important processes. We have observed that the cytoplasmic distribution of dynamin changes dramatically in fibroblasts that have been stimulated to undergo migration with a motagen/hormone. In quiescent cells, dynamin 2 (Dyn 2) associates predominantly with clathrin-coated vesicles at the plasma membrane and the Golgi apparatus. Upon treatment with PDGF to induce cell migration, dynamin becomes markedly associated with membrane ruffles and lamellipodia. Biochemical and morphological studies using antibodies and GFP-tagged dynamin demonstrate an interaction with cortactin. Cortactin is an actin-binding protein that contains a well defined SH3 domain. Using a variety of biochemical methods we demonstrate that the cortactin-SH3 domain associates with the proline-rich domain (PRD) of dynamin. Functional studies that express wild-type and mutant forms of dynamin and/or cortactin in living cells support these in vitro observations and demonstrate that an increased expression of cortactin leads to a significant recruitment of endogenous or expressed dynamin into the cell ruffle. Further, expression of a cortactin protein lacking the interactive SH3 domain (CortDeltaSH3) significantly reduces dynamin localization to the ruffle. Accordingly, transfected cells expressing Dyn 2 lacking the PRD (Dyn 2(aa)DeltaPRD) sequester little of this protein to the cortactin-rich ruffle. Interestingly, these mutant cells are viable, but display dramatic alterations in morphology. This change in shape appears to be due, in part, to a striking increase in the number of actin stress fibers. These findings provide the first demonstration that dynamin can interact with the actin cytoskeleton to regulate actin reorganization and subsequently cell shape. PMID- 11018065 TI - Surviving ischemia: adaptive responses mediated by hypoxia-inducible factor 1. PMID- 11018066 TI - Protective responses in the ischemic myocardium. PMID- 11018067 TI - Calpain 10: the first positional cloning of a gene for type 2 diabetes? PMID- 11018068 TI - Latency and viral persistence in HIV-1 infection. PMID- 11018069 TI - Matrix metalloproteinases: not-so-innocent bystanders in heart failure. PMID- 11018070 TI - VEGF enhances angiogenesis and promotes blood-brain barrier leakage in the ischemic brain. AB - VEGF is a secreted mitogen associated with angiogenesis and is also a potent vascular permeability factor. The biological role of VEGF in the ischemic brain remains unknown. This study was undertaken to investigate whether VEGF enhances cerebral microvascular perfusion and increases blood-brain barrier (BBB) leakage in the ischemic brain. Using magnetic resonance imaging (MRI), three-dimensional laser-scanning confocal microscope, and functional neurological tests, we measured the effects of administrating recombinant human VEGF(165) (rhVEGF(165)) on angiogenesis, functional neurological outcome, and BBB leakage in a rat model of focal cerebral embolic ischemia. Late (48 hours) administration of rhVEGF(165) to the ischemic rats enhanced angiogenesis in the ischemic penumbra and significantly improved neurological recovery. However, early postischemic (1 hour) administration of rhVEGF(165) to ischemic rats significantly increased BBB leakage, hemorrhagic transformation, and ischemic lesions. Administration of rhVEGF(165) to ischemic rats did not change BBB leakage and cerebral plasma perfusion in the contralateral hemisphere. Our results indicate that VEGF can markedly enhance angiogenesis in the ischemic brain and reduce neurological deficits during stroke recovery and that inhibition of VEGF at the acute stage of stroke may reduce the BBB permeability and the risk of hemorrhagic transformation after focal cerebral ischemia. PMID- 11018071 TI - Genetic characterization of rebounding HIV-1 after cessation of highly active antiretroviral therapy. AB - Despite prolonged treatment with highly active antiretroviral therapy (HAART), infectious HIV-1 continues to replicate and to reside latently in resting memory CD4(+) T lymphocytes, creating a major obstacle to HIV-1 eradication. It is therefore not surprising to observe a prompt viral rebound after discontinuation of HAART. The nature of the rebounding virus, however, remains undefined. We now report on the genetic characterization of rebounding viruses in eight patients in whom plasma viremia was undetectable throughout about 3 years of HAART. Taking advantage of the extensive length polymorphism in HIV-1 env, we found that in five patients who did not show HIV-1 replication during treatment, the rebound virus was identical to those isolated from the latent reservoir. In three other patients, two of whom had been free of plasma viremia but had showed some residual viral replication, the rebound virus was genetically different from the latent reservoir virus, corresponding instead to minor viral variants detected during the course of treatment in lymphoid tissues. We conclude that in cases with apparent complete HIV-1 suppression by HAART, viral rebound after cessation of therapy could have originated from the activation of virus from the latent reservoir. In patients with incomplete suppression by chemotherapy, however, the viral rebound is likely triggered by ongoing, low-level replication of HIV-1, perhaps occurring in lymphoid tissues. PMID- 11018072 TI - Peroxisome proliferator-activated receptor gamma coactivator-1 promotes cardiac mitochondrial biogenesis. AB - Cardiac mitochondrial function is altered in a variety of inherited and acquired cardiovascular diseases. Recent studies have identified the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator-1 (PGC 1) as a regulator of mitochondrial function in tissues specialized for thermogenesis, such as brown adipose. We sought to determine whether PGC-1 controlled mitochondrial biogenesis and energy-producing capacity in the heart, a tissue specialized for high-capacity ATP production. We found that PGC-1 gene expression is induced in the mouse heart after birth and in response to short term fasting, conditions known to increase cardiac mitochondrial energy production. Forced expression of PGC-1 in cardiac myocytes in culture induced the expression of nuclear and mitochondrial genes involved in multiple mitochondrial energy-transduction/energy-production pathways, increased cellular mitochondrial number, and stimulated coupled respiration. Cardiac-specific overexpression of PGC-1 in transgenic mice resulted in uncontrolled mitochondrial proliferation in cardiac myocytes leading to loss of sarcomeric structure and a dilated cardiomyopathy. These results identify PGC-1 as a critical regulatory molecule in the control of cardiac mitochondrial number and function in response to energy demands. PMID- 11018073 TI - Disruption of the myocardial extracellular matrix leads to cardiac dysfunction. AB - MMP activity with disruption of structural collagen has been implicated in the pathophysiology of dilated cardiomyopathy. To examine the role of this enzyme in cardiac function, a transgenic mouse was created that constitutively expressed human collagenase (MMP-1) in the heart. At 6 months of age, these animals demonstrated compensatory myocyte hypertrophy with an increase in the cardiac collagen concentration due to elevated transcription of type III collagen. Chronic myocardial expression of MMP-1 produced loss of cardiac interstitial collagen coincident with a marked deterioration of systolic and diastolic function at 12 months of age. This is the first animal model demonstrating that direct disruption of the extracellular matrix in the heart reproduces the changes observed in the progression of human heart failure. PMID- 11018074 TI - NADPH oxidase-derived free radicals are key oxidants in alcohol-induced liver disease. AB - In North America, liver disease due to alcohol consumption is an important cause of death in adults, although its pathogenesis remains obscure. Despite the fact that resident hepatic macrophages are known to contribute to early alcohol induced liver injury via oxidative stress, the exact source of free radicals has remained a mystery. To test the hypothesis that NADPH oxidase is the major source of oxidants due to ethanol, we used p47(phox) knockout mice, which lack a critical subunit of this major source of reactive oxygen species in activated phagocytes. Mice were treated with ethanol chronically, using a Tsukamoto-French protocol, for 4 weeks. In wild-type mice, ethanol caused severe liver injury via a mechanism involving gut-derived endotoxin, CD14 receptor, production of electron spin resonance-detectable free radicals, activation of the transcription factor NF-kappaB, and release of cytotoxic TNF-alpha from activated Kupffer cells. In NADPH oxidase-deficient mice, neither an increase in free radical production, activation of NF-kappaB, an increase in TNF-alpha mRNA, nor liver pathology was observed. These data strongly support the hypothesis that free radicals from NADPH oxidase in hepatic Kupffer cells play a predominant role in the pathogenesis of early alcohol-induced hepatitis by activating NF-kappaB, which activates production of cytotoxic TNF-alpha. PMID- 11018075 TI - Complete antithrombin deficiency in mice results in embryonic lethality. AB - Antithrombin is a plasma protease inhibitor that inhibits thrombin and contributes to the maintenance of blood fluidity. Using targeted gene disruption, we investigated the role of antithrombin in embryogenesis. Mating mice heterozygous for antithrombin gene (ATIII) disruption, ATIII(+/-), yielded the expected Mendelian distribution of genotypes until 14.5 gestational days (gd). However, approximately 70% of the ATIII(-/-) embryos at 15.5 gd and 100% at 16.5 gd had died and showed extensive subcutaneous hemorrhage. Histological examination of those embryos revealed extensive fibrin(ogen) deposition in the myocardium and liver, but not in the brain or lung. Furthermore, no apparent fibrin(ogen) deposition was detected in the extensive hemorrhagic region, suggesting that fibrinogen might be decreased due to consumptive coagulopathy and/or liver dysfunction. These findings suggest that antithrombin is essential for embryonic survival and that it plays an important role in regulation of blood coagulation in the myocardium and liver. PMID- 11018076 TI - Enhanced intestinal transepithelial antigen transport in allergic rats is mediated by IgE and CD23 (FcepsilonRII). AB - We previously reported that active sensitization of rats resulted in the appearance of a unique system for rapid and specific antigen uptake across intestinal epithelial cells. The current studies used rats sensitized to horseradish peroxidase (HRP) to define the essential components of this antigen transport system. Sensitization of rats to HRP stimulated increased HRP uptake into enterocytes (significantly larger area of HRP-containing endosomes) and more rapid transcellular transport compared with rats sensitized to an irrelevant protein or naive control rats. Whole serum but not IgE-depleted serum from sensitized rats was able to transfer the enhanced antigen transport phenomenon. Immunohistochemistry demonstrated that sensitization induced expression of CD23, the low-affinity IgE receptor (FcepsilonRII), on epithelial cells. The number of immunogold-labeled CD23 receptors on the enterocyte microvillous membrane was significantly increased in sensitized rats and was subsequently reduced after antigen challenge when CD23 and HRP were localized within the same endosomes. Finally, pretreatment of tissues with luminally added anti-CD23 antibody significantly inhibited both antigen transport and the hypersensitivity reaction. Our results provide evidence that IgE antibodies bound to low-affinity receptors on epithelial cells are responsible for the specific and rapid nature of this novel antigen transport system. PMID- 11018077 TI - Chronic alcohol ingestion induces osteoclastogenesis and bone loss through IL-6 in mice. AB - To investigate the role of IL-6 in alcohol-mediated osteoporosis, we measured a variety of bone remodeling parameters in wild-type (il6(+/+)) or IL-6 gene knockout (il6(-/-)) mice that were fed either control or ethanol liquid diets for 4 months. In the il6(+/+) mice, ethanol ingestion decreased bone mineral density, as determined by dual-energy densitometry; decreased cancellous bone volume and trabecular width and increased trabecular spacing and osteoclast surface, as determined by histomorphometry of the femur; increased urinary deoxypyridinolines, as determined by ELISA; and increased CFU-GM formation and osteoclastogenesis as determined ex vivo in bone marrow cell cultures. In contrast, ethanol ingestion did not alter any of these parameters in the il6(-/-) mice. Ethanol increased receptor activator of NF-kappaB ligand (RANKL) mRNA expression in the bone marrow of il6(+/+) but not il6(-/-) mice. Additionally, ethanol decreased several osteoblastic parameters including osteoblast perimeter and osteoblast culture calcium retention in both il6(+/+) and il6(-/-) mice. These findings demonstrate that ethanol induces bone loss through IL-6. Furthermore, they suggest that IL-6 achieves this effect by inducing RANKL and promoting CFU-GM formation and osteoclastogenesis. PMID- 11018078 TI - Dominantly inherited hyperinsulinism caused by a mutation in the sulfonylurea receptor type 1. AB - ATP-sensitive potassium channels play a major role in linking metabolic signals to the exocytosis of insulin in the pancreatic beta cell. These channels consist of two types of protein subunit: the sulfonylurea receptor SUR1 and the inward rectifying potassium channel Kir6.2. Mutations in the genes encoding these proteins are the most common cause of congenital hyperinsulinism (CHI). Since 1973, we have followed up 38 pediatric CHI patients in Finland. We reported previously that a loss-of-function mutation in SUR1 (V187D) is responsible for CHI of the most severe cases. We have now identified a missense mutation, E1506K, within the second nucleotide binding fold of SUR1, found heterozygous in seven related patients with CHI and in their mothers. All patients have a mild form of CHI that usually can be managed by long-term diazoxide treatment. This clinical finding is in agreement with the results of heterologous coexpression studies of recombinant Kir6.2 and SUR1 carrying the E1506K mutation. Mutant K(ATP) channels were insensitive to metabolic inhibition, but a partial response to diazoxide was retained. Five of the six mothers, two of whom suffered from hypoglycemia in infancy, have developed gestational or permanent diabetes. Linkage and haplotype analysis supported a dominant pattern of inheritance in a large pedigree. In conclusion, we describe the first dominantly inherited SUR1 mutation that causes CHI in early life and predisposes to later insulin deficiency. PMID- 11018079 TI - Processing of chromogranin A by plasmin provides a novel mechanism for regulating catecholamine secretion. AB - Chromogranin A (CgA) is the major soluble protein in the core of catecholamine storage vesicles and is also distributed widely in secretory vesicles throughout the neuroendocrine system. CgA contains the sequences for peptides that modulate catecholamine release, but the proteases responsible for the release of these bioactive peptides from CgA have not been established. We show here that the major fibrinolytic enzyme, plasmin, can cleave CgA to form a series of large fragments as well as small trichloroacetic acid-soluble peptides. Peptides generated by plasmin-mediated cleavage of CgA significantly inhibited nicotinic cholinergic stimulation of catecholamine release from PC12 cells and primary bovine adrenal chromaffin cells. We also show that the zymogen, plasminogen, as well as tissue plasminogen activator bind saturably and with high capacity to catecholaminergic (PC12) cells. Occupancy of cell surface binding sites promoted the cleavage of CgA by plasmin. Positive and negative modulation of the local cellular fibrinolytic system resulted in substantial alterations in catecholamine release. These results suggest that catecholaminergic cells express binding sites that localize fibrinolytic molecules on their surfaces to promote plasminogen activation and proteolytic processing of CgA in the environment into which CgA is secreted to generate peptides which may regulate neuroendocrine secretion. Interactions between CgA and plasmin(ogen) define a previously unrecognized autocrine/paracrine system that may have a dramatic impact upon catecholamine secretion. PMID- 11018080 TI - A calpain-10 gene polymorphism is associated with reduced muscle mRNA levels and insulin resistance. AB - Previous linkage studies in Mexican-Americans localized a major susceptibility locus for type 2 diabetes, NIDDM1, to chromosome 2q. This evidence for linkage to type 2 diabetes was recently found to be associated with a common G-->A polymorphism (UCSNP-43) within the CAPN10 gene. The at-risk genotype was homozygous for the UCSNP-43 G allele. In the present study among Pima Indians, the UCSNP-43 G/G genotype was not associated with an increased prevalence of type 2 diabetes. However, Pima Indians with normal glucose tolerance, who have a G/G genotype at UCSNP-43, were found to have decreased rates of postabsorptive and insulin-stimulated glucose turnover that appear to result from decreased rates of glucose oxidation. In addition, G/G homozygotes were found to have reduced CAPN10 mRNA expression in their skeletal muscle. A decreased rate of insulin-mediated glucose turnover, or insulin resistance, is one mechanism by which the polymorphism in CAPN10 may increase susceptibility to type 2 diabetes mellitus in older persons. PMID- 11018081 TI - Use of an antibody to target geldanamycin. PMID- 11018082 TI - Response rates, survival, and chemotherapy trials. PMID- 11018083 TI - Model aims to reduce discrepancy between phase II, phase III trials. PMID- 11018084 TI - How does a drug get to phase III trials? PMID- 11018085 TI - Surveys identify barriers to participation in clinical trials. PMID- 11018086 TI - Stat bite: Reasons for not participating in cancer clinical trials. PMID- 11018087 TI - Alternative therapies getting notice through Best Case Series Program. PMID- 11018088 TI - What goes into an NCI best case series? PMID- 11018089 TI - Gender differences in cancer that don't make sense--or do they? PMID- 11018091 TI - Consensus meeting to focus on adjuvant therapy for breast cancer PMID- 11018090 TI - Journal to encourage analysis by sex/ethnicity. PMID- 11018092 TI - Role of the heat shock response and molecular chaperones in oncogenesis and cell death. AB - Exposure of cells to conditions of environmental stress-including heat shock, oxidative stress, heavy metals, or pathologic conditions, such as ischemia and reperfusion, inflammation, tissue damage, infection, and mutant proteins associated with genetic diseases-results in the inducible expression of heat shock proteins that function as molecular chaperones or proteases. Molecular chaperones are a class of proteins that interact with diverse protein substrates to assist in their folding, with a critical role during cell stress to prevent the appearance of folding intermediates that lead to misfolded or otherwise damaged molecules. Consequently, heat shock proteins assist in the recovery from stress either by repairing damaged proteins (protein refolding) or by degrading them, thus restoring protein homeostasis and promoting cell survival. The events of cell stress and cell death are linked, such that molecular chaperones induced in response to stress appear to function at key regulatory points in the control of apoptosis. On the basis of these observations-and on the role of molecular chaperones in the regulation of steroid aporeceptors, kinases, caspases, and other protein remodeling events involved in chromosome replication and changes in cell structure-it is not surprising that the heat shock response and molecular chaperones have been implicated in the control of cell growth. In this review, we address some of the molecular and cellular events initiated by cell stress-the interrelationships between stress signaling, cell death, and oncogenesis-and chaperones as potential targets for cancer diagnosis and treatment. PMID- 11018093 TI - Immunoconjugates of geldanamycin and anti-HER2 monoclonal antibodies: antiproliferative activity on human breast carcinoma cell lines. AB - BACKGROUND: HER2 is a membrane receptor whose overexpression is strongly associated with poor prognosis in breast carcinomas. Inhibition of HER2 activity can reduce tumor growth, which led to the development of Herceptin, an anti-HER2 monoclonal antibody (MAb) that is already in clinical use. However, the objective response rate to Herceptin monotherapy is quite low. HER2 activity can also be inhibited by the highly cytotoxic antibiotic geldanamycin (GA). However, GA is not used clinically because of its adverse toxicity. Our purpose was to enhance the inhibitory activity of anti-HER2 MAb by coupling it to GA. METHODS: We synthesized 17-(3-aminopropylamino)GA (17-APA-GA) and conjugated it to the anti HER2 MAb e21, to form e21 : GA. The noninternalizing anti-HER2 MAb AE1 was used as a control. Internalization assays and western blot analyses were used to determine whether the anti-HER2 MAbs and their immunoconjugates were internalized into HER2-expressing cells and reduced HER2 levels. All statistical tests were two-sided. RESULTS: The immunoconjugate e21 : GA inhibited the proliferation of HER2-overexpressing cell lines better than unconjugated e21 (concentration required for 50% inhibition = 40 versus 1650 microg/mL, respectively). At 15 microg/mL, e21 : GA reduced HER2 levels by 86% within 16 hours, whereas unconjugated e21, 17-APA-GA, or AE1 : GA reduced HER2 levels by only 20%. These effects were not caused by release of 17-APA-GA from the immunoconjugate because immunoconjugates containing [(3)H]GA were stable in serum at 37 degrees C. Furthermore, e21 : GA did not significantly inhibit proliferation of the adult T cell leukemia cell line HuT102, which is HER2 negative yet highly sensitive to GA. CONCLUSIONS: Our findings suggest that conjugating GA to internalizing MAbs enhances the inhibitory effect of the MAbs. This approach might also be applied in cellular targeting via growth factors and may be of clinical interest. PMID- 11018094 TI - Health outcomes after prostatectomy or radiotherapy for prostate cancer: results from the Prostate Cancer Outcomes Study. AB - BACKGROUND: Radical prostatectomy and external beam radiotherapy are the two major therapeutic options for treating clinically localized prostate cancer. Because survival is often favorable regardless of therapy, treatment decisions may depend on other therapy-specific health outcomes. In this study, we compared the effects of two treatments on urinary, bowel, and sexual functions and on general health-related quality-of-life outcomes over a 2-year period following initial treatment. METHODS: A diverse cohort of patients aged 55-74 years who were newly diagnosed with clinically localized prostate cancer and received either radical prostatectomy (n = 1156) or external beam radiotherapy (n = 435) were included in this study. A propensity score was used to balance the two treatment groups because they differed in some baseline characteristics. This score was used in multivariable cross-sectional and longitudinal regression analyses comparing the treatment groups. All statistical tests were two-sided. RESULTS: Almost 2 years after treatment, men receiving radical prostatectomy were more likely than men receiving radiotherapy to be incontinent (9.6% versus 3.5%; P:<.001) and to have higher rates of impotence (79.6% versus 61.5%; P:<.001), although large, statistically significant declines in sexual function were observed in both treatment groups. In contrast, men receiving radiotherapy reported greater declines in bowel function than did men receiving radical prostatectomy. All of these differences remained after adjustments for propensity score. The treatment groups were similar in terms of general health-related quality of life. CONCLUSIONS: There are important differences in urinary, bowel, and sexual functions over 2 years after different treatments for clinically localized prostate cancer. In contrast to previous reports, these outcome differences reflect treatment delivered to a heterogeneous group of patients in diverse health care settings. These results provide comprehensive and representative information about long-term treatment complications to help guide and inform patients and clinicians about prostate cancer treatment decisions. PMID- 11018095 TI - Serum C-peptide, insulin-like growth factor (IGF)-I, IGF-binding proteins, and colorectal cancer risk in women. AB - BACKGROUND: Leading a Western lifestyle, being overweight, and being sedentary are associated with an increased risk of colorectal cancer. Recent theories propose that the effects of these risk factors may be mediated by increases in circulating insulin levels and in the bioactivity of insulin-like growth factor (IGF)-I. To test this hypothesis, we conducted a case-control study nested within a cohort of 14 275 women in New York. METHODS: We used blood samples that had been obtained from these women from March 1985 through June 1991 and stored in a biorepository. C-peptide (a marker for insulin secretion), IGF-I, and IGF-binding proteins (IGFBPs)-1, -2, and -3 were assayed in the serum of 102 women who subsequently developed colorectal cancer and 200 matched control subjects. Logistic regression was used to relate cancer risk to these peptide levels, by adjustment for other risk factors. All statistical tests used are two-sided. RESULTS: Colorectal cancer risk increased with increasing levels of C-peptide (P:(trend) =.001), up to an odds ratio (OR) of 2. 92 (95% confidence interval [CI] = 1.26-6.75) for the highest versus the lowest quintiles, after adjustment for smoking. For colon cancer alone (75 case subjects and 146 control subjects), ORs increased up to 3.96 (95% CI = 1.49-10.50; P:(trend) <.001) for the highest versus the lowest quintiles. A statistically significant decrease in colorectal cancer risk was observed for increasing levels of IGFBP-1 (P:(trend) =.02; OR in the upper quintile = 0.48 [95% CI = 0.23-1. 00]), as well as for the highest quintile of IGFBP-2 levels (P:(trend) =.06; OR = 0.38 [95% CI = 0.15-0.94]). Colorectal cancer risk showed a modest but statistically nonsignificant positive association with levels of IGF-I and was statistically significantly increased for the highest quintile of IGFBP-3 (OR = 2.46 [95% CI = 1. 09-5.57]). CONCLUSIONS: Chronically high levels of circulating insulin and IGFs associated with a Western lifestyle may increase colorectal cancer risk, possibly by decreasing IGFBP-1 and increasing the bioactivity of IGF-I. PMID- 11018096 TI - A model to select chemotherapy regimens for phase III trials for extensive-stage small-cell lung cancer. AB - BACKGROUND: Many more phase II studies have favorable outcomes than the subsequent phase III trials. We used historical data from phase II and phase III studies for patients with extensive-stage small-cell lung cancer (SCLC) to generate a statistical model to provide assistance in selecting chemotherapy regimens from phase II studies for subsequent use in phase III randomized studies. METHODS: Information from 21 phase III trials for patients with extensive-stage SCLC initiated during the period from 1972 through 1990 was reviewed to identify those that were preceded by phase II studies of the same regimen. We used data from all the trial pairs to develop a statistical model in which the number of patients, the median survival of patients, and the number of deaths observed in the phase II trial are used to estimate the statistical power of the subsequent phase III trial. All statistical tests were two-sided. RESULTS: Nine phase II studies were identified that preceded phase III trials of the same regimen. The regimens from two phase II studies with the greatest expected power in the phase III trial (0. 62 and 0.58) both demonstrated significantly prolonged survival when compared with standard treatment in subsequent phase III trials (P<. 001 and P =.002, respectively). The regimens from six of the other phase II studies, for which the median power expected in the phase III trial was 0.28 (range, 0.19-0.52), showed no difference when compared with standard treatment in a phase III trial. CONCLUSIONS: Phase II studies for particular regimens that have an expected power of greater than 0.55 provide a reasonable basis for proceeding with a phase III trial. PMID- 11018097 TI - Gastric dysplasia and gastric cancer: Helicobacter pylori, serum vitamin C, and other risk factors. AB - BACKGROUND: Gastric cancer is generally thought to arise through a series of gastric mucosal changes, but the determinants of the precancerous lesions are not well understood. To identify such determinants, we launched a follow-up study in 1989-1990 among 3433 adults in Linqu County, China, a region with very high rates of gastric cancer. METHODS: Data on cigarette smoking, alcohol consumption, and other characteristics of the participants were obtained by interview in 1989 1990, when an initial endoscopy was taken. At study entry, antibodies to Helicobacter pylori were assayed in 2646 adults (77% of people screened), and levels of serum micronutrients were measured in approximately 450 adults. Follow up endoscopic and histopathologic examinations were conducted in 1994. Antibodies to H. pylori, levels of serum micronutrients, and other baseline characteristics were compared between subjects whose condition showed progression to dysplasia or gastric cancer from study entry to 1994 and subjects with no change or with regression of their lesions over the same time frame. All P: values are two sided. RESULTS: The presence of H. pylori at baseline was associated with an increased risk of progression to dysplasia or gastric cancer (odds ratio [OR] = 1.8; 95% confidence interval [CI] = 1.2-2.6). The risk of progression to dysplasia or gastric cancer also was moderately increased with the number of years of cigarette smoking. In contrast, the risk of progression was decreased by 80% (OR = 0.2; 95% CI = 0.1-0.7) among subjects with baseline ascorbic acid levels in the highest tertile compared with those in the lowest tertile, and there was a slightly elevated risk in those individuals with higher levels of alpha-tocopherol. CONCLUSIONS: H. pylori infection, cigarette smoking, and low levels of dietary vitamin C may contribute to the progression of precancerous lesions to gastric cancer in this high-risk population. PMID- 11018098 TI - Primary afferent depolarization produced in Adelta and C fibres by glutamate spillover? New ways to look at old things. PMID- 11018099 TI - NO and the regulation of VSOACs. PMID- 11018100 TI - How to avoid running on empty. PMID- 11018101 TI - Stable, polarised, functional expression of Kir1.1b channel protein in Madin Darby canine kidney cell line. AB - 1. The family of Kir1.1 (ROMK) channel proteins constitute a secretory pathway for potassium in principal cells of cortical collecting duct and thick ascending limb of Henle's loop. Mutations in Kir1.1 account for some types of Bartter's syndrome. 2. Here we report that stable transfection of Kir1.1b (ROMK2) in Madin Darby canine kidney (MDCK) cell line results in expression of inwardly rectifying K+ currents and transmonolayer electrical and transport properties appropriate to Kir1.1 function. When grown on permeable supports, transfected monolayers secreted K+ into the apical solution. This secretion was inhibited by application of barium to the apical membrane, or by reduction in expression temperature from 37 to 26 C. However, whole-cell voltage clamp electrophysiology showed that K+ conductance was higher in cells expressing Kir1.1b at 26C. 3. To investigate this further, Kir1.1b was tagged with (EGFP), a modification that did not affect channel activity. Protein synthesis was inhibited with cycloheximide. Spectrofluorimetry was used to compare protein degradation at 37 and 26 C. The increased level of Kir1.1b at the plasma membrane at 26 C was due to an increase in protein stability. 4. Confocal microscopic investigation of EGFP-Kir1. 1b fluorescence in transfected cells showed that the channel protein was targeted to the apical domain of the cell. 5. These results demonstrate that Kir1.1b is capable of appropriate trafficking and function in MDCK cell lines at physiological temperatures. In addition, expression of Kir1.1b in MDCK cell lines provides a useful and convenient tool for the study of functional activity and targeting of secretory K+ channels. PMID- 11018102 TI - Dual modulation of swelling-activated chloride current by NO and NO donors in rabbit portal vein myocytes. AB - 1. The effects of authentic NO and the NO donor S-nitroso-N-acetylpenicillamine (SNAP) on swelling-activated chloride currents (Iswell) were investigated in freshly dispersed rabbit portal vein smooth muscle cells. Iswell was recorded with the perforated patch configuration of the whole-cell patch clamp technique. 2. In approximately 50 % of cells NO and SNAP inhibited the amplitude of Iswell by about 45 % in a voltage-independent manner. Iswell was also inhibited by an inhibitor of NO-sensitive guanylate cyclase (1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ) and by KT5823, an inhibitor of cGMP-dependent protein kinase. 3. In other cells both NO and SNAP enhanced Iswell by about 40 % in a voltage-independent manner. A similar increase was produced by application of the cell-permeable cGMP analogue 8-bromo-guanosine 3', 5'-cyclic monophosphate (8-Br cGMP). However, 8-Br-cGMP had no effect on current amplitude in cells pre-treated with KT5823. In contrast 8-Br-cGMP increased the amplitude of Iswell in cells which had been pre-treated with ODQ. 4. SNAP also modulated Iswell recorded in the conventional whole-cell configuration with internal solutions containing 10 mM EGTA to rule out any contribution from Ca2+-activated Cl- currents. 5. These data suggest that the amplitude of Iswell can be enhanced by NO via a cGMP dependent phosphorylation and inhibited by NO in a cGMP-independent manner. PMID- 11018103 TI - Effect of intracellular pH on spontaneous Ca2+ sparks in rat ventricular myocytes. AB - 1. A fall of intracellular pH (pHi) typically depresses cardiac contractility. Among the many mechanisms underlying this depression, an inhibitory effect of acidosis upon the sarcoplasmic reticulum (SR) Ca2+ release channel has been predicted, but not so far demonstrated in the intact cardiac myocyte. In the present work, pHi was manipulated experimentally while confocal imaging was used to record spontaneous 'Ca2+ sparks' (local SR Ca2+ release events) in rat isolated myocytes loaded with the fluorescent Ca2+ indicator fluo-3. In other experiments, whole cell (global) pHi or [Ca2+]i was measured by microfluorimetry (using, respectively, intracellular carboxy SNARF-1 and indo-1). 2. Reducing pHi (i) increased whole cell intracellular [Ca2+] transients induced either electrically or by addition of caffeine, whereas (ii) it decreased spontaneous Ca2+ spark frequency. Conversely, raising pHi increased spontaneous Ca2+ spark frequency. 3. Blocking sarcolemmal Ca2+ influx with 10 mM Ni2+, or reducing external pH by 1.0 unit, had no effect on the pHi-dependent changes in spontaneous Ca2+ spark frequency. 4. Decreasing pHi over the range 7.78-7.20, decreased Ca2+ spark frequency exponentially as a function of pHi, with frequency declining by approximately 33 % for a 0.2 unit fall in pHi. In contrast, over the same pHi range, Ca2+ spark amplitude was unaffected. Intracellular acidosis produced a slight slowing of Ca2+ spark relaxation. 5. The results indicate that, in the intact myocyte, a reduced pHi decreases the probability of opening of the SR Ca2+ release channel. This phenomenon may contribute to the negative inotropic effects of acidosis. PMID- 11018104 TI - Zn2+ current is mediated by voltage-gated Ca2+ channels and enhanced by extracellular acidity in mouse cortical neurones. AB - 1. Mammalian neuronal voltage-gated Ca2+ channels have been implicated as potential mediators of membrane permeability to Zn2+. We tested directly whether voltage-gated Ca2+ channels can flux Zn2+ in whole-cell voltage-clamp recordings from cultured murine cortical neurones. 2. In the presence of extracellular Zn2+ and no Na+, K+, or other divalent cations, a small, non-inactivating, voltage gated inward current was observed exhibiting a current-voltage relationship characteristic of high-voltage activated (HVA) Ca2+ channels. Inward current was detectable at Zn2+ levels as low as 50 microM, and both the amplitude and voltage sensitivity of the current depended upon Zn2+ concentration. This Zn2+ current was sensitive to blockade by Gd3+ and nimodipine and, to a lesser extent, by omega-conotoxin GVIA. 3. Zn2+ could permeate Ca2+ channels in the presence of Ca2+ and other physiological cations. Inward currents recorded with 2 mM Ca2+ were attenuated by Zn2+ (IC50 = 210 microM), and currents recorded with Zn2+ were unaffected by up to equimolar Ca2+ concentrations. Furthermore, the Zn2+ selective fluorescent dye Newport Green revealed a depolarisation-activated, nimodipine-sensitive Zn2+ influx into cortical neurones that were bathed in a physiological extracellular solution plus 300 microM ZnCl2. 4. Surprisingly, while lowering extracellular pH suppressed HVA Ca2+ currents, Zn2+ current amplitude was affected oppositely, varying inversely with pH with an apparent pK of 7.4. The acidity-induced enhancement of Zn2+ current was associated with a positive shift in reversal potential but no change in the kinetics or voltage sensitivity of channel activation. 5. These results provide evidence that L- and N-type voltage-gated Ca2+ channels can mediate Zn2+ entry into cortical neurones and that this entry may be enhanced by extracellular acidity. PMID- 11018105 TI - Exogenous Ca2+-ATPase isoform effects on Ca2+ transients of embryonic chicken and neonatal rat cardiac myocytes. AB - 1. Sarco-endoplasmic reticulum Ca2+-ATPase from fast skeletal (SERCA1) or cardiac muscle (SERCA2a) was expressed in embryonic chicken and neonatal rat cardiac myocytes by adenovirus vectors, with c-myc tags on both constructs to compare expression and distinguish exogenous from endogenous SERCA2a in myocytes. 2. Expression of the two isoforms was similar (approximately 3-fold higher than endogenous SERCA). However, SERCA1 activity was 2-fold greater than SERCA2a activity, due to intrinsic differences in turnover rates. Activation of both exogenous SERCA isoforms by Ca2+ was displaced to slightly lower [Ca2+], suggesting that the overexpressed isoforms were independent of phospholamban. In fact, phospholamban and calsequestrin expression were unchanged. 3. Decay time constants of cytosolic Ca2+ transients from cells overexpressing SERCA1 were reduced by 30-40 % and half-widths by 10-15 % compared to controls. SERCA2a overexpression produced much less acceleration of transients in chick than in rat, and less acceleration than SERCA1 overexpression in either species. There was no significant change in resting [Ca2+], peak amplitudes, or in the amount of Ca2+ releasable by caffeine from overexpression of either SERCA isoform. However, the amplitudes of the transients increased with SERCA1 overexpression when pacing frequency limited refilling of the sarcoplasmic reticulum. 4. It is concluded that total SERCA transport velocity has a primary effect on the decay phase of transients. Transport velocity is affected by SERCA isoform turnover rate, temperature, and/or SERCA copy number. PMID- 11018107 TI - Voltage-gated Ca2+ channels and intracellular Ca2+ release regulate exocytosis in identified rat corticotrophs. AB - 1. The patch clamp technique was used in conjunction with a fluorescent Ca2+ indicator (indo-1, or indo-1FF) to measure simultaneously cytosolic Ca2+ concentration ([Ca2+]i) and exocytosis (changes in membrane capacitance) in single, identified rat corticotrophs. 2. Exocytosis could be stimulated by extracellular Ca2+ entry (via voltage-gated Ca2+ channels). A train of depolarizations could exhaust the pool of readily releasable granules and the pool replenished with a time constant of 42 s (at 22-25 C). 3. Recordings from cells with 0.5 mM intracellular cAMP showed that the amplitude of the depolarization-triggered exocytosis, the Ca2+ sensitivity of exocytosis, as well as the rate of replenishment of the readily releasable pool, were similar to the controls. 4. Exocytosis could also be stimulated by intracellular Ca2+ release from the inositol 1,4, 5-trisphosphate (IP3)-sensitive store (via flash photolysis of caged IP3). At comparable [Ca2+]i, extracellular Ca2+ entry and intracellular Ca2+ release had similar efficacy in triggering exocytosis. 5. The rate of exocytosis triggered via depolarization or intracellular Ca2+ release was much faster than that triggered via uniform elevation of [Ca2+]i (Ca2+ dialysis or flash photolysis of caged Ca2+). 6. The above findings suggest that both intracellular Ca2+ release and voltage-gated extracellular Ca2+ entry generate a spatial Ca2+ gradient, such that the local [Ca2+] near the exocytic sites was approximately 3-fold higher than the mean cytosolic [Ca2+]. However, neither cAMP nor the spatial Ca2+ gradient generated during depolarization could account for the high efficacy of corticotropin-releasing hormone (CRH) in stimulating adrenocorticotropic hormone (ACTH) secretion from corticotrophs. PMID- 11018106 TI - G protein modulation of recombinant P/Q-type calcium channels by regulators of G protein signalling proteins. AB - 1. Fast synaptic transmission is triggered by the activation of presynaptic Ca2+ channels which can be inhibited by Gbetagamma subunits via G protein-coupled receptors (GPCR). Regulators of G protein signalling (RGS) proteins are GTPase accelerating proteins (GAPs), which are responsible for >100-fold increases in the GTPase activity of G proteins and might be involved in the regulation of presynaptic Ca2+ channels. In this study we investigated the effects of RGS2 on G protein modulation of recombinant P/Q-type channels expressed in a human embryonic kidney (HEK293) cell line using whole-cell recordings. 2. RGS2 markedly accelerates transmitter-mediated inhibition and recovery from inhibition of Ba2+ currents (IBa) through P/Q-type channels heterologously expressed with the muscarinic acetylcholine receptor M2 (mAChR M2). 3. Both RGS2 and RGS4 modulate the prepulse facilitation properties of P/Q-type Ca2+ channels. G protein reinhibition is accelerated, while release from inhibition is slowed. These kinetics depend on the availability of G protein alpha and betagamma subunits which is altered by RGS proteins. 4. RGS proteins unmask the Ca2+ channel beta subunit modulation of Ca2+ channel G protein inhibition. In the presence of RGS2, P/Q-type channels containing the beta2a and beta3 subunits reveal significantly altered kinetics of G protein modulation and increased facilitation compared to Ca2+ channels coexpressed with the beta1b or beta4 subunit. PMID- 11018108 TI - Voltage-gated transient outward currents in neurons with different firing patterns in rat superior colliculus. AB - 1. We investigated the electrophysiological properties of transient outward currents (TOCs) in neurons with different firing patterns, regular-spiking, fast spiking and late-spiking neurons, in the intermediate layer (SGI) of the superior colliculus using the whole-cell patch clamp technique in slice preparations obtained from young rats (post-natal days 17-22). 2. Analysis of inactivation kinetics and normalized amplitude revealed that TOCs in regular-and fast-spiking neurons had fast inactivation kinetics (decay time constants (mean +/- s.e.m.) of 13.8 +/- 1.5 and 11.4 +/- 1.2 ms, respectively) and low current densities (36.6 +/- 3.3 and 32.1 +/- 4. 9 pA pF-1, respectively). TOCs in late-spiking neurons, on the other hand, displayed a wide range of both inactivation kinetics (36.7 +/- 2.4 ms, with a range from 11.3 to 147.8 ms) and current density (54. 0 +/- 2.9 pA pF-1, with a range from 9.8 to 131.2 pA pF-1). 3. In regular-, fast- and late spiking neurons having TOCs with slow time constants (> 50 ms, class II late spiking neurons), the TOCs were sensitive to 4-aminopyridine (4-AP), with IC50 values of 2.9, 2.4 and 1.2 mM, respectively. In late-spiking neurons having TOCs with fast decay time constants (< 30 ms, class I late-spiking neurons), the TOCs were composed of at least two 4-AP-sensitive components (IC50 values of 0.2 microM and 3.6 mM). 4. Class I late-spiking neurons displayed non-inactivating outward currents which were highly sensitive to 4-AP. They changed their firing patterns to the regular-spiking mode, not only in response to low concentrations of 4-AP (< 50 microM), but also in response to dendrotoxin (200 nM), suggesting that non-inactivating outward currents contribute to the late-spiking property. However, the components of TOCs which were highly sensitive to 4-AP were also sensitive to dendrotoxin. These results suggest that both or either of the two currents contribute to the late-spiking property of class I late-spiking neurons. 5. Although class II late-spiking neurons also displayed non-inactivating outward currents, the late-spiking property was not abolished by low concentrations of 4 AP and dendrotoxin. They changed to a regular firing pattern in response to a high concentration of 4-AP (5 mM), suggesting that TOCs contribute to late spiking property of class II late-spiking neurons. 6. The results suggest that TOCs with different properties contribute to the different firing patterns of SGI neurons. PMID- 11018109 TI - Facilitation of plateau potentials in turtle motoneurones by a pathway dependent on calcium and calmodulin. AB - 1. The involvement of intracellular calcium and calmodulin in the modulation of plateau potentials in motoneurones was investigated using intracellular recordings from a spinal cord slice preparation. 2. Chelation of intracellular calcium with BAPTA-AM or inactivation of calmodulin with W-7 or trifluoperazine reduced the amplitude of depolarization-induced plateau potentials. Inactivation of calmodulin also inhibited facilitation of plateau potentials by activation of group I metabotropic glutamate receptors or muscarinic receptors. 3. In low sodium medium and in the presence of tetraethylammonium and tetrodotoxin, calcium action potentials evoked by depolarization were followed by a short hyperpolarization ascribed to the calcium-activated non-selective cationic current (ICAN) and by a dihydropyridine-sensitive afterdepolarization. The amplitude of the afterdepolarization depended on the number of calcium spikes and was mediated by L-type calcium channels. 4. The dihydropyridine-sensitive afterdepolarization induced by calcium spikes was reduced by blockade of calmodulin. 5. It is proposed that plateau potentials in spinal motoneurones are facilitated by activation of a calcium-calmodulin-dependent pathway. PMID- 11018110 TI - Dorsal root potential produced by a TTX-insensitive micro-circuitry in the turtle spinal cord. AB - 1, The mechanisms underlying the dorsal root potential (DRP) were studied in transverse slices of turtle spinal cord. DRPs were evoked by stimulating one filament in a dorsal root and were recorded from another such filament. 2. The DRP evoked at supramaximal stimulus intensity was reduced but not eliminated after blockade of GABAA receptors. The remaining component was eliminated by blocking NMDA and AMPA receptors. 3. The DRP was reduced but not eliminated after blockade of AMPA receptors. The early component of the remaining DRP was dependent on GABAA receptors and the residual component on NMDA receptors. 4. The DRP was reduced but not eliminated by TTX. GABAA, NMDA and AMPA receptors contributed to the generation of the TTX-insensitive DRP. The early component of the DRP in the presence of TTX depended on GABAA receptor activation, and the late component mainly on the activation of NMDA receptors. 5. Our results show that part of the DRP is generated by a TTX-resistant, probably non-spiking micro circuit with separate components mediated by GABA and glutamate. PMID- 11018111 TI - Differential control of GABA release at synapses from distinct interneurons in rat hippocampus. AB - 1. Paired recordings from monosynaptically connected CA3 interneurons and pyramidal cells of rat hippocampal slice cultures were used to compare the modulation of GABA release at synapses from distinct interneurons. 2. The group II metabotropic glutamate receptor (mGluR) agonist (2S,2'R,3'R)-2-(2',3' dicarboxylcyclopropyl) glycine (DCG-IV, 5 muM) reduced the amplitude of IPSPs originating from stratum radiatum but not stratum oriens interneurons. In contrast, the GABAB receptor agonist (-)baclofen (10 muM) reduced the amplitude of unitary IPSPs elicited by all interneurons. 3. IPSPs mediated by stratum oriens interneurons were unaffected by the N-type calcium channel blocker omega conotoxin MVIIA (1 muM) but were suppressed by the P/Q-type blocker omega agatoxin IVA (200 nM). In contrast, IPSPs mediated by stratum radiatum interneurons were abolished by omega-conotoxin MVIIA. 4. Transmission dynamics were different at synapses from the two groups of interneurons. IPSPs mediated by stratum oriens interneurons showed marked paired-pulse depression (PPD) at intervals of 50 400 ms. IPSPs mediated by stratum radiatum interneurons showed paired-pulse facilitation (PPF) at 50 ms and PPD at longer intervals. 5. The amplitude of unitary IPSPs from all interneurons was unaffected by the GABAB receptor antagonist CGP52432 (2 muM) as was PPD at both 50 and 400 ms intervals. However, CGP52432 did reduce PPD of extracellularly evoked IPSPs. 6. Our results show that two groups of inhibitory synapses impinging onto CA3 pyramidal cells can be distinguished according to their dynamic and modulatory properties. PMID- 11018112 TI - Relationship between the time course of the afterhyperpolarization and discharge variability in cat spinal motoneurones. AB - 1. We elicited repetitive discharges in cat spinal motoneurones by injecting noisy current waveforms through a microelectrode to study the relationship between the time course of the motoneurone's afterhyperpolarization (AHP) and the variability in its spike discharge. Interspike interval histograms were used to estimate the interval death rate, which is a measure of the instantaneous probability of spike occurrence as a function of the time since the preceding spike. It had been previously proposed that the death rate can be used to estimate the AHP trajectory. We tested the accuracy of this estimate by comparing the AHP trajectory predicted from discharge statistics to the measured AHP trajectory of the motoneurone. 2. The discharge statistics of noise-driven cat motoneurones shared a number of features with those previously reported for voluntarily activated human motoneurones. At low discharge rates, the interspike interval histograms were often positively skewed with an exponential tail. The standard deviation of the interspike intervals increased with the mean interval, and the plots of standard deviation versus the mean interspike interval generally showed an upward bend, the onset of which was related to the motoneurone's AHP duration. 3. The AHP trajectories predicted from the interval death rates were generally smaller in amplitude (i.e. less hyperpolarized) than the measured AHP trajectories. This discrepancy may result from the fact that spike threshold varies during the interspike interval, so that the distance to threshold at a given time depends upon both the membrane trajectory and the spike threshold trajectory. Nonetheless, since the interval death rate is likely to reflect the instantaneous distance to threshold during the interspike interval, it provides a functionally relevant measure of fluctuations in motoneurone excitability during repetitive discharge. PMID- 11018113 TI - A role for C-protein in the regulation of contraction and intracellular Ca2+ in intact rat ventricular myocytes. AB - 1. C-protein is a major component of muscle thick filaments whose function is unknown. We have examined for the first time the role of the regulatory binding domain of C-protein in modulating contraction and intracellular Ca2+ concentration ([Ca2+]i) in intact cardiac myocytes. 2. Rat ventricular myocytes were reversibly permeabilised with the pore-forming toxin streptolysin O. Myosin S2 (which binds to the regulatory domain of C-protein) was introduced into cells during permeabilisation to compete with the endogenous C-protein-thick filament interaction. 3. Introduction of S2 into myocytes increased contractility by approximately 30%, significantly lengthened the time to peak of the contraction and the time to half-relaxation, but had no effect on [Ca2+]i transient amplitude. 4. Our data are consistent with increased myofilament Ca2+ sensitivity when there is reduced binding of C-protein to myosin near the head-tail junction. 5. We propose that the effects of introducing S2 into intact cardiac cells can be equated with the consequences of selectively phosphorylating C-protein in vivo, and that the regulation of contraction by C-protein is mediated by the effects of crossbridge cycling on the Ca2+ affinity of troponin C. PMID- 11018114 TI - Muscle contractions induce interleukin-6 mRNA production in rat skeletal muscles. AB - 1. The present study explored the hypothesis that interleukin-6 (IL-6) might be locally produced in response to skeletal muscle contractions and whether the production might reflect the type of muscle contraction performed. Rats were anaesthetized and the calf muscles of one limb were stimulated electrically for concentric or eccentric contractions (4 x 10 contractions with 1 min of rest between the 4 series, 100 Hz). The contralateral muscles served as unstimulated controls. The mRNA levels for IL-6, the glucose transport protein GLUT-4 and beta actin in the rat muscles (white and red gastrocnemius and soleus) were quantified by quantitative competitive RT-PCR. 2. The IL-6 mRNA level, measured 30 min after the stimulation, increased after both eccentric and concentric contractions and there were no significant differences in IL-6 mRNA levels between the different muscle fibre types. No significant increase in IL-6 mRNA level was seen in the unstimulated contralateral muscle fibres. 3. No increase in GLUT-4 mRNA level was detected, indicating that the increase in IL-6 mRNA level was not due to general changes in transcription. 4. We conclude that IL-6 is locally produced after muscle contraction, with no significant differences between different muscle fibre types. This local production of IL-6 is not due to general changes in transcription, since no changes in the level of GLUT-4 mRNA were found. The fact that increased IL-6 mRNA levels were seen after both concentric and eccentric contractions indicates that the production of IL-6 is not solely due to muscle damage, seen primarily after eccentric exercise. PMID- 11018115 TI - Fatty acid-induced cholecystokinin secretion and changes in intracellular Ca2+ in two enteroendocrine cell lines, STC-1 and GLUTag. AB - 1. Fatty acid-induced cholecystokinin (CCK) secretion in humans and from the enteroendocrine cell line STC-1 depends critically on acyl chain length. 2. Therefore we have characterized the relationship between acyl chain length and the potency of the fatty acid to induce CCK secretion and changes in intracellular Ca2+ concentration ([Ca2+]i) in two enteroendocrine cell lines (STC 1 and GLUTag). We found that the potency of the fatty acid was directly proportional to its chain length and therefore inversely proportional to its solubility. 3. In both cell types, the fatty acid-induced rise in [Ca2+]i in response to decanoic acid (C10), dodecanoic acid (C12) and tetradecanoic acid (C14) was significantly reduced in Ca2+-free medium and largely blocked by nicardipine. Intracellular stores also contributed to the overall shape of the [Ca2+]i peak. Thus all the fatty acids tested caused the release of Ca2+ from stores and influx of extracellular Ca2+, presumably through L-type calcium channels. 4. To probe the site of fatty acid action, we studied the distribution of 14C-labelled dodecanoic acid. This label was rapidly and irreversibly accumulated by both cell types, where it became concentrated about 20-fold. Confocal microscopy of a fluorescent analogue of dodecanoic acid clearly demonstrated that it entered the cytosol and was not merely partitioning in the cell membrane. These data indicate that an intracellular action for fatty acid induced CCK secretion cannot be eliminated. 5. Dodecanoic acid itself, and not a metabolite, is the agent responsible for triggering Ca2+ entry since a non metabolizable form of dodecanoic acid (2-bromododecanoic acid) was also capable of inducing a rise in [Ca2+]i in both cell types. 6. In conclusion, the rise in [Ca2+]i in STC-1 and GLUTag cells evoked by medium- to long-chain fatty acids results from the triggering of a specific signalling pathway. Whether triggering occurs through activation of a membrane-bound receptor or at an intracellular site remains to be clarified. PMID- 11018116 TI - Acute effects of bradykinin on cerebral microvascular permeability in the anaesthetized rat. AB - 1. The permeability response to acutely applied bradykinin and [des-Arg9] bradykinin on single cerebral venular capillaries has been investigated using the low molecular mass fluorescent dyes Lucifer Yellow and Sulforhodamine B with the single vessel occlusion technique. 2. When bradykinin was applied repeatedly for up to 2 h, the permeability increase was small and reversible for concentrations that ranged from 5 nM to 50 microM. 3. The logEC50 of the permeability response to bradykinin was -5.3 +/- 0.15 (logM; mean +/- s.e.m.). This was reduced to 6.37 +/- 0.24 with the angiotensin-converting enzyme inhibitor captopril, to 6.33 +/- 0.19 with the neutral endopeptidase inhibitor phosphoramidon and to -7.3 +/- 0.20 with captopril and phosphoramidon combined. 4. The permeability response to bradykinin was blocked by the bradykinin B2 receptor antagonist HOE 140, by inhibition of the Ca2+-independent phospholipase A2, by the scavenging of free radicals, or by inhibition of both cyclo-oxygenase and lipoxygenase in combination. Block of Ca2+ entry channels with SKF 96365 had no effect on the response. 5. Application of [des-Arg9]-bradykinin also increased permeability over the concentration range 5 nM to 50 microM, with a logEC50 of -5.6 +/- 0. 37. This response was not affected by free radical scavenging, but was completely blocked by the histamine H2 receptor blocker cimetidine. 6. These results imply that the acute permeability response to bradykinin is mediated via the release of arachidonic acid, which is acted on by cyclo-oxygenase and lipoxygenase resulting in the formation of free radicals, and that the response to [des-Arg9]-bradykinin is mediated via histamine. PMID- 11018117 TI - Inhibition of basolateral cAMP permeability in the toad urinary bladder. AB - 1. The effect of sulphonylurea drugs on hydrosmotic flow across toad urinary bladder epithelium was re-evaluated in the present study. Glibenclamide, added to the basolateral medium, significantly enhanced the osmotic flow induced by low doses of antidiuretic hormone (ADH) or forskolin (FK), while it inhibited the effect of exogenous cyclic adenosine monophosphate (cAMP) or its non-hydrolysable bromo derivative, 8-Br-cAMP, added to the basolateral medium. These opposite effects of glibenclamide on the transepithelial osmotic flow can be explained by a reduction of cAMP permeability across the basolateral membrane of the epithelium. The decrease in cAMP permeability leads, according to the direction of the cAMP gradient, to firstly an enhanced osmotic flow when cAMP is generated intracellularly by addition of ADH and FK, glibenclamide reducing cAMP exit from the cell, and secondly a decreased osmotic flow in response to cAMP (and 8-Br cAMP) added to the basolateral medium, glibenclamide inhibiting, in this case, their entry into the cell. 2. The demonstration that glibenclamide actually inhibits the basolateral cAMP permeability rests on the fact that firstly it decreases the release of cAMP into the basolateral medium by about 40 %, at each concentration of ADH or forskolin tested, secondly it increases the cAMP content of paired hemibladders incubated in the presence of ADH or FK, when intracellular degradation was prevented by phosphodiesterase inhibition, and thirdly it decreases also the uptake of basolateral 8-Br-[3H]cAMP into paired toad hemibladders. 3. Taken together, the present data demonstrate that glibenclamide inhibits the toad urinary bladder basolateral membrane permeability to cAMP, most probably by a direct interaction with a membrane protein not yet indentified but distinct from the sulphonylurea receptor. PMID- 11018118 TI - Estimation of aqueous distributional spaces in the dual perfused rat liver. AB - 1. The aim of this study was to estimate the aqueous distributional spaces of the liver as a function of the route of input: portal vein (PV) versus hepatic artery (HA). 2. Studies were performed in the situ single (PV) and dual (PV-HA) perfused rat liver (n = 6-10) using Krebs bicarbonate buffer at constant PV (12 ml min-1) and HA (3 ml min-1) flow rates. An impulse input-output response technique was employed, varying the route of input, using non-labelled erythrocytes (intravascular marker), 125I-albumin and [14C]sucrose (extracellular markers), and [14C]urea and 3H2O (total water markers) as the reference indicators. 3. Distributional spaces were estimated using two different methods, namely standard and specific. The standard method was applied to hepatic outflow data obtained from the single PV perfused liver. The specific method was used when operating in the dual perfused mode to provide an estimate of the excess space perfused solely by the HA input. Specific spaces, interstitial and intracellular volumes, were estimated by difference. 4. The results were evaluated by means of visual inspection of the outflow profiles and comparison of the distributional spaces. Different hepatic effluent profiles obtained as a function of the route of input indicated that these two inputs did not completely mix within the liver. Estimates of the distributional spaces supported this observation, and further suggested that the arterial input perfuses 9-12 % more hepatic tissue than the venous input. 5. The knowledge obtained from the existence of a specific arterial space can be extended to help make predictions about the fate of an eliminated compound following arterial administration. Any difference between the HA and PV in terms of hepatic recovery could be attributed to this excess space and its enzyme density. PMID- 11018119 TI - Ontogenic and nutritionally induced changes in fetal metabolism in the horse. AB - 1. Using the Fick principle and tracer methodology, the metabolic rates of chronically catheterized fetal foals (n = 24) were measured at different gestational ages during the second half of gestation and in response to maternal fasting for 36 h in late gestation (n = 4, term approximately 335 days). 2. Absolute rates of umbilical blood flow, fetal glucose utilization and umbilical uptake of oxygen (O2) and glucose increased from mid-gestation to 300 days and then plateauxed until term. The absolute rate of umbilical lactate uptake was significant after, but not before, 280 days of gestation. Weight specific rates of umbilical uptake and fetal utilization of glucose decreased progressively throughout the second half of gestation. 3. Weight specific rates of CO2 production from glucose carbon were also inversely correlated with gestational age. Umbilical lactate uptake per kilogram of fetus was lower after 320 days than between 281 and 300 days. In contrast, no gestational trends were observed in the weight specific rates of fetal O2 uptake and urea production. Glucose production by the fetal horse was negligible, even very close to term. 4. Maternal fasting for 36 h reduced the rate of umbilical uptake and utilization of glucose production but had no effect on the rates of endogenous glucose production and umbilical uptake of oxygen and lactate by the horse fetus. 5. The observations show that fetal metabolism is highly dependent on glucose in the horse compared to the sheep in mid- and late gestation. Different species therefore adopt different strategies in meeting the nutritional demands of the growing fetus during the second half of gestation. PMID- 11018120 TI - Isoform-specific and exercise intensity-dependent activation of 5'-AMP-activated protein kinase in human skeletal muscle. AB - 1. 5'-AMP-activated protein kinase (AMPK) has been suggested to play a key role in the regulation of metabolism in skeletal muscle. AMPK is activated in treadmill-exercised and electrically stimulated rodent muscles. Whether AMPK is activated during exercise in humans is unknown. 2. We investigated the degree of activation and deactivation of alpha-isoforms of AMPK during and after exercise. Healthy human subjects performed bicycle exercise on two separate occasions at either a low ( approximately 50% maximum rate of O2 uptake (VO2,max) for 90 min) or a high ( approximately 75% VO2,max for 60 min) intensity. Biopsies from the vastus lateralis muscle were obtained before and immediately after exercise, and after 3 h of recovery. 3. We observed a 3- to 4-fold activation of the alpha2 AMPK isoform immediately after high intensity exercise, whereas no activation was observed after low intensity exercise. The activation of alpha2-AMPK was totally reversed 3 h after exercise. In contrast, alpha1-AMPK was not activated during either of the two exercise trials. 4. The in vitro AMP dependency of alpha2-AMPK was significantly greater than that of alpha1-AMPK ( approximately 3- vs. approximately 2-fold). 5. We conclude that in humans activation of alpha2-AMPK during exercise is dependent upon exercise intensity. The stable activation of alpha2-AMPK, presumably due to the activation of an upstream AMPK kinase, is compatible with a role for this kinase complex in the regulation of skeletal muscle metabolism during exercise, whereas the lack of stable alpha1-AMPK activation makes this kinase complex a less likely candidate. PMID- 11018121 TI - Discharge behaviour of single motor units during maximal voluntary contractions of a human toe extensor. AB - 1. While it is known that the average firing rate of a population of motoneurones declines with time during a maximal voluntary contraction, at least for many muscles, it is not known how the firing patterns of individual motoneurones adapt with fatigue. To address this issue we used tungsten microelectrodes to record spike trains (mean +/- s.e.m., 183 +/- 27 spikes per train; range, 100-782 spikes) from 26 single motor units in extensor hallucis longus during sustained (60-180 s) maximal dorsiflexions of the big toe in seven human subjects. 2. Long spike trains were recorded from 13 units during the first 30 s of a maximal voluntary contraction (mean train duration, 9.6 +/- 1.2 s; range, 3.6-21.9 s) and from 13 units after 30 s (mean train duration, 16.6 +/- 3.7 s; range, 7.1-58.1 s). Maximal isometric force generated by the big toe declined to 78.3 +/- 6.3 % of its control level by 60-90 s and to 39.5 +/- 1.4 % of control by 120-150 s. Despite this substantial fatigue, mean firing rates did not change significantly over time, declining only slightly from 15.8 +/- 0.7 Hz in the first 30 s to 14.0 +/- 0.5 Hz by 60-90 s and 13.6 +/- 0.3 Hz by 120-150 s. 3. To assess fatigue related adaptation in discharge frequency and variability of individual motor units, each spike train was divided into 2-15 equal segments containing at least 50 interspike intervals. Discharge variability was measured from the coefficient of variation (s.d. /mean) in the interspike intervals, with the s.d. being calculated using a floating mean of 19 consecutive intervals. Adaptation was computed as the average change in firing rate or variability that would occur for each 1 s of activity. There were no systematic changes in either firing rate or variability with time. 4. We conclude that single motoneurones supplying the extensor hallucis longus, a muscle comprised primarily of slow twitch muscle units, show little adaptation in firing with fatigue, suggesting that a progressive reduction in firing rate is not an invariable consequence of the fatigue associated with sustained maximal voluntary contractions. PMID- 11018122 TI - The life and times of a clinical microbiologist. PMID- 11018123 TI - Role of cytotoxic T lymphocytes in Epstein-Barr virus-associated diseases. AB - Adaptation of persistent infection within the cells of the immune system is a unique characteristic of gamma herpes viruses. A classic example of this is Epstein-Barr virus (EBV), which may have co-evolved with Homo sapiens over millions of years, thus achieving a balance between viral persistence and immune control. In this review, we present an overview of virus and the host immune system interactions that regulate the life-long host-virus relationship in healthy virus carriers and EBV-associated diseases. Extensive analysis of cytotoxic T lymphocyte-mediated immune responses in healthy virus carriers has revealed unique mechanisms used by EBV to maintain a benign persistent state in vivo. On the other hand, this relationship in EBV-associated diseases favors the escape of the virus from the hostile effects of the immune response. This escape is achieved by either down-regulating the expression of highly immunogenic antigens of the virus or by direct modulation of the host cytotoxic T lymphocyte response by virus-encoded proteins. PMID- 11018124 TI - Biofilm formation as microbial development. AB - Biofilms can be defined as communities of microorganisms attached to a surface. It is clear that microorganisms undergo profound changes during their transition from planktonic (free-swimming) organisms to cells that are part of a complex, surface-attached community. These changes are reflected in the new phenotypic characteristics developed by biofilm bacteria and occur in response to a variety of environmental signals. Recent genetic and molecular approaches used to study bacterial and fungal biofilms have identified genes and regulatory circuits important for initial cell-surface interactions, biofilm maturation, and the return of biofilm microorganisms to a planktonic mode of growth. Studies to date suggest that the planktonic-biofilm transition is a complex and highly regulated process. The results reviewed in this article indicate that the formation of biofilms serves as a new model system for the study of microbial development. PMID- 11018125 TI - Microbiological safety of drinking water. AB - Emerging pathogens in drinking water have become increasingly important during the decade. These include newly-recognized pathogens from fecal sources such as Cryptosporidium parvum, Campylobacter spp., and rotavirus, as well as pathogens that are able to grow in water distribution systems, like Legionella spp., mycobacteria, and aeromonads. To perform a risk analysis for the pathogens in drinking water, it is necessary to understand the ecology of these organisms. The ecology of the drinking-water distribution system has to be evaluated in detail, especially the diversity and physiological properties of water bacteria. The interactions between water bacteria and (potential) pathogens in such diverse habitats as free water and biofilms are essential for the survival or growth of hygienically relevant organisms in drinking water. Results of epidemiological studies together with ecological data are the basis for effective resource protection, water treatment, and risk assessment. PMID- 11018126 TI - The adaptative mechanisms of Trypanosoma brucei for sterol homeostasis in its different life-cycle environments. AB - Bloodstream forms of Trypanosoma brucei do not synthesize sterols de novo and therefore cannot survive in medium devoid of lipoproteins. Growth of parasites is essentially supported by receptor-mediated endocytosis of low-density lipoproteins (LDLs), which carry phospholipids and cholesteryl esters. These lipids are released from internalized LDL after apoprotein B-100 is degraded by acidic thiol-proteases in the endolysosomal apparatus and then metabolized, as in mammalian cells. The LDL receptor is recycled and its expression is regulated by the sterol stores. Documented pharmacological and immunological interferences with LDL receptor-mediated lipid supply to the bloodstream forms are summarized, and the potential for new approaches to fight against these parasites is evaluated. In contrast to bloodstream forms, cultured procyclic forms can acquire sterols from both exogenous (lipoprotein endocytosis) and endogenous (biosynthesis of ergosterol) sources. The rate-limiting steps of both endocytosis (surface LDL receptor expression) and biosynthesis (3-hydroxy-3-methylglutaryl coenzyme A reductase activity) are regulated by the cellular content of sterol. These two pathways thus complement each other to yield a balanced sterol supply, which demonstrates adaptative capacities to survive in totally different environments and fine regulatory mechanisms of sterol homeostasis. PMID- 11018127 TI - The development of genetic tools for dissecting the biology of malaria parasites. AB - Plasmodium parasites are haploid unicellular organisms that cause malaria. In the last decade, transfection systems have been developed for both human and animal model species of Plasmodium, providing a broad range of genetic tools for the study of malaria parasite biology. Transient transfection has been used to provide insight into the regulation of gene expression by Plasmodium spp. The development of stable transfection technologies has provided the opportunity to express transgenes in Plasmodium spp., as well as elucidate the function of proteins by disrupting, modifying, or replacing the genes encoding them. These genetic tools represent an important breakthrough for malaria research and will significantly contribute to our understanding of the biology of the parasite. However, further developments in this technology are still required, especially because the full genome sequence of the major human malaria parasite Plasmodium falciparum will shortly be available. Ultimately, the biological information obtained through genetic manipulation of Plasmodium spp. will facilitate a more rational approach to vaccine and drug design. PMID- 11018128 TI - Nucleic acid transport in plant-microbe interactions: the molecules that walk through the walls. AB - Many microbes "genetically invade" plants by introducing DNA or RNA molecules into the host cells. For example, plant viruses transport their genomes between host cells, whereas Agrobacterium spp. transfer T-DNA to the cell nucleus and integrate it into the plant DNA. During these events, the transported nucleic acids must negotiate several barriers, such as plant cell walls, plasma membranes, and nuclear envelopes. This review describes the microbial and host proteins that participate in cell-to-cell transport and nuclear import of nucleic acids during infection by plant viruses and Agrobacterium spp. Possible molecular mechanisms by which these transport processes occur are discussed. PMID- 11018129 TI - Phytoplasma: phytopathogenic mollicutes. AB - During the past decade, research has yielded new knowledge about the plant and insect host ranges, geographical distribution, and phylogenetic relationships of phytoplasmas, and a taxonomic system has emerged in which distinct phytoplasmas are named as separate "Candidatus phytoplasma species." In large part, this progress has resulted from the development and use of molecular methods to detect, identify, and classify phytoplasmas. While these advances continue, research has recently begun on the phytoplasma genome, how phytoplasmas cause disease, the role of mixed phytoplasmal infections in plant diseases, and molecular/genetic phenomena that underlie symptom development in plants. These and other recent advances are laying the foundation for future progress in understanding the mechanisms of phytoplasma pathogenicity, organization of the phytoplasma genome, evolution of new phytoplasma strains and emergence of new diseases, bases of insect transmissibility and specificity of transmission, and plant gene expression in response to phytoplasmal infection, as well as the design of novel approaches to achieve effective control of phytoplasmal diseases. PMID- 11018130 TI - Root nodulation and infection factors produced by rhizobial bacteria. AB - Rhizobia are soil bacteria that can engage in a symbiosis with leguminous plants that produces nitrogen-fixing root nodules. This symbiosis is based on specific recognition of signal molecules, which are produced by both the bacterial and plant partners. In this review, recognition factors from the bacterial endosymbionts are discussed, with particular attention to secreted and cell surface glycans. Glycans that are discussed include the Nod factors, the extracellular polysaccharides, the lipopolysaccharides, the K-antigens, and the cyclic glucans. Recent advances in the understanding of the biosynthesis, secretion, and regulation of production of these glycans are reviewed, and their functions are compared with glycans produced by other bacteria, such as plant pathogens. PMID- 11018131 TI - ALGINATE LYASE: review of major sources and enzyme characteristics, structure function analysis, biological roles, and applications. AB - Alginate lyases, characterized as either mannuronate (EC 4.2.2.3) or guluronate lyases (EC 4.2.2.11), catalyze the degradation of alginate, a complex copolymer of alpha-L-guluronate and its C5 epimer beta-D-mannuronate. Lyases have been isolated from a wide range of organisms, including algae, marine invertebrates, and marine and terrestrial microorganisms. This review catalogs the major characteristics of these lyases, the methods for analyzing these enzymes, as well as their biological roles. Analysis of primary sequence data identifies some markedly conserved motifs that should help elucidate functional domains. Information about the three-dimensional structure of a mannuronate lyase from Sphingomonas sp., combined with various mutagenesis studies, has identified residues that are important for catalytic activity in several lyases. Characterization of alginate lyases will enhance and expand the use of these enzymes to engineer novel alginate polymers for applications in various industrial, agricultural, and medical fields. In this review, we explore both past and present applications of this important enzyme and discuss its future prospects. PMID- 11018132 TI - Interim report on genomics of Escherichia coli. AB - We present a summary of recent progress in understanding Escherichia coli K-12 gene and protein functions. New information has come both from classical biological experimentation and from using the analytical tools of functional genomics. The content of the E. coli genome can clearly be seen to contain elements acquired by horizontal transfer. Nevertheless, there is probably a large, stable core of >3500 genes that are shared among all E. coli strains. The gene-enzyme relationship is examined, and, in many cases, it exhibits complexity beyond a simple one-to-one relationship. Also, the E. coli genome can now be seen to contain many multiple enzymes that carry out the same or closely similar reactions. Some are similar in sequence and may share common ancestry; some are not. We discuss the concept of a minimal genome as being variable among organisms and obligatorily linked to their life styles and defined environmental conditions. We also address classification of functions of gene products and avenues of insight into the history of protein evolution. PMID- 11018133 TI - Oral microbial communities: biofilms, interactions, and genetic systems. AB - Oral microbial-plaque communities are biofilms composed of numerous genetically distinct types of bacteria that live in close juxtaposition on host surfaces. These bacteria communicate through physical interactions called coaggregation and coadhesion, as well as other physiological and metabolic interactions. Streptococci and actinomyces are the major initial colonizers of the tooth surface, and the interactions between them and their substrata help establish the early biofilm community. Fusobacteria play a central role as physical bridges that mediate coaggregation of cells and as physiological bridges that promote anaerobic microenvironments which protect coaggregating strict anaerobes in an aerobic atmosphere. New technologies for investigating bacterial populations with 16S rDNA probes have uncovered previously uncultured bacteria and have offered an approach to in situ examination of the spatial arrangement of the participant cells in oral-plaque biofilms. Flow cells with saliva-coated surfaces are particularly useful for studies of biofilm formation and observation. The predicted sequential nature of colonization of the tooth surface by members of different genera can be investigated by using these new technologies and imaging the cells in situ with confocal scanning laser microscopy. Members of at least seven genera now can be subjected to genetic studies owing to the discovery of gene-transfer systems in these genera. Identification of contact-inducible genes in streptococci offers an avenue to explore bacterial responses to their environment and leads the way toward understanding communication among inhabitants of a multispecies biofilm. PMID- 11018134 TI - Roles of the glutathione- and thioredoxin-dependent reduction systems in the Escherichia coli and saccharomyces cerevisiae responses to oxidative stress. AB - The glutathione- and thioredoxin-dependent reduction systems are responsible for maintaining the reduced environment of the Escherichia coli and Saccharomyces cerevisiae cytosol. Here we examine the roles of these two cellular reduction systems in the bacterial and yeast defenses against oxidative stress. The transcription of a subset of the genes encoding glutathione biosynthetic enzymes, glutathione reductases, glutaredoxins, thioredoxins, and thioredoxin reductases, as well as glutathione- and thioredoxin-dependent peroxidases is clearly induced by oxidative stress in both organisms. However, only some strains carrying mutations in single genes are hypersensitive to oxidants. This is due, in part, to the redundant effects of the gene products and the overlap between the two reduction systems. The construction of strains carrying mutations in multiple genes is helping to elucidate the different roles of glutathione and thioredoxin, and studies with such strains have recently revealed that these two reduction systems modulate the activities of the E. coli OxyR and SoxR and the S. cerevisiae Yap1p transcriptional regulators of the adaptive responses to oxidative stress. PMID- 11018135 TI - Recent developments in molecular genetics of Candida albicans. AB - The frequency of opportunistic infections caused by the fungus Candida albicans is very high and is expected to continue to increase as the number of immunocompromised patients rises. Research initiatives to study the biology of this organism and elucidate its pathogenic determinants have therefore expanded significantly during the last 5-10 years. The past few years have also brought continuous improvement in the techniques to study gene function by gene inactivation and by regulated gene expression and to study gene expression and protein localization by using gene reporter systems. As steadily more genomic sequence information from this human fungal pathogen becomes available, we are entering a new era in antimicrobial research. However, many of the currently available molecular genetics tools are poorly adapted to a genome-wide functional analysis in C. albicans, and further development of these tools is hampered by the asexual and diploid nature of this organism. This review outlines recent advances in the development of molecular tools for functional analysis in C. albicans and summarizes current knowledge about the genomic and genetic variability of this important human fungal pathogen. PMID- 11018136 TI - Functional modulation of Escherichia coli RNA polymerase. AB - The promoter recognition specificity of Escherichia coli RNA polymerase is modulated by replacement of the sigma subunit in the first step and by interaction with transcription factors in the second step. The overall differentiated state of approximately 2000 molecules of the RNA polymerase in a single cell can be estimated after measurement of both the intracellular concentrations and the RNA polymerase-binding affinities for all seven species of the sigma subunit and 100-150 transcription factors. The anticipated impact from this line of systematic approach is that the prediction of the expression hierarchy of approximately 4000 genes on the E. coli genome can be estimated. PMID- 11018137 TI - Bacterial virulence gene regulation: an evolutionary perspective. AB - Coevolution between bacteria and their plant or animal hosts determines characteristics of the interaction, the bacterial virulence genes involved, and the regulatory systems controlling expression of virulence genes. The long standing association between Salmonellae and their animal hosts has resulted in the acquisition by Salmonella subspecies of a variety of virulence genes and the evolution of complex regulatory networks. The particular repertoire of virulence genes acquired by different Salmonella enterica subspecies and the regulatory systems that control them dictate subspecies-specific infection characteristics. Although the association between Vibrio cholerae and humans appears to be more recent, to reflect a simpler pathogenic strategy, and to involve fewer virulence genes than that of Salmonellae, complex virulence-regulatory networks have nonetheless evolved. In contrast, there is no evidence for acquisition of virulence genes by horizontal gene transfer in bordetellae, and their virulence regulon is less complex in overall structure than those of salmonellae and Vibrio cholerae. In Bordetellae, subspecies-specific differences in pathogenic strategy appear to result from differential gene expression within and across Bordetella subspecies. PMID- 11018138 TI - Legionella pneumophila pathogesesis: a fateful journey from amoebae to macrophages. AB - Legionella pneumophila first commanded attention in 1976, when investigators from the Centers for Disease Control and Prevention identified it as the culprit in a massive outbreak of pneumonia that struck individuals attending an American Legion convention (). It is now clear that this gram-negative bacterium flourishes naturally in fresh water as a parasite of amoebae, but it can also replicate within alveolar macrophages. L. pneumophila pathogenesis is discussed using the following model as a framework. When ingested by phagocytes, stationary phase L. pneumophila bacteria establish phagosomes which are completely isolated from the endosomal pathway but are surrounded by endoplasmic reticulum. Within this protected vacuole, L. pneumophila converts to a replicative form that is acid tolerant but no longer expresses several virulence traits, including factors that block membrane fusion. As a consequence, the pathogen vacuoles merge with lysosomes, which provide a nutrient-rich replication niche. Once the amino acid supply is depleted, progeny accumulate the second messenger guanosine 3',5' bispyrophosphate (ppGpp), which coordinates entry into the stationary phase with expression of traits that promote transmission to a new phagocyte. A number of factors contribute to L. pneumophila virulence, including type II and type IV secretion systems, a pore-forming toxin, type IV pili, flagella, and numerous other factors currently under investigation. Because of its resemblance to certain aspects of Mycobacterium, Toxoplasma, Leishmania, and Coxiella pathogenesis, a detailed description of the mechanism used by L. pneumophila to manipulate and exploit phagocyte membrane traffic may suggest novel strategies for treating a variety of infectious diseases. Knowledge of L. pneumophila ecology may also inform efforts to combat the emergence of new opportunistic macrophage pathogens. PMID- 11018139 TI - The disease spectrum of Helicobacter pylori: the immunopathogenesis of gastroduodenal ulcer and gastric cancer. AB - Helicobacter pylori is a gram-negative bacterium that resides under microaerobic conditions in a neutral microenvironment between the mucus and the superficial epithelium of the stomach. From this site, it stimulates cytokine production by epithelial cells that recruit and activate immune and inflammatory cells in the underlying lamina propria, causing chronic, active gastritis. Although epidemiological evidence shows that infection generally occurs in children, the inflammatory changes progress throughout life. H. pylori has also been recognized as a pathogen that causes gastroduodenal ulcers and gastric cancer. These more severe manifestations of the infection usually occur later in life and in a minority of infected subjects. To intervene and protect those who might be at greatest risk of the more severe disease outcomes, it is of great interest to determine whether bacterial, host, or environmental factors can be used to predict these events. To date, several epidemiological studies have attempted to define the factors affecting the transmission of H. pylori and the expression of gastroduodenal disease caused by this infection. Many other laboratories have focused on identifying bacterial factors that explain the variable expression of clinical disease associated with this infection. An alternative hypothesis is that microorganisms that cause lifelong infections can ill afford to express virulence factors that directly cause disease, because the risk of losing the host is too great. Rather, we propose that gastroduodenal disease associated with H. pylori infection is predominantly a result of inappropriately regulated gastric immune responses to the infection. In this model, the interactions between the immune/inflammatory response, gastric physiology, and host repair mechanisms would dictate the disease outcome in response to infection. PMID- 11018140 TI - Pathogenicity islands and the evolution of microbes. AB - Virulence factors of pathogenic bacteria (adhesins, toxins, invasins, protein secretion systems, iron uptake systems, and others) may be encoded by particular regions of the prokaryotic genome termed pathogenicity islands. Pathogenicity islands were first described in human pathogens of the species Escherichia coli, but have recently been found in the genomes of various pathogens of humans, animals, and plants. Pathogenicity islands comprise large genomic regions [10-200 kilobases (kb) in size] that are present on the genomes of pathogenic strains but absent from the genomes of nonpathogenic members of the same or related species. The finding that the G+C content of pathogenicity islands often differs from that of the rest of the genome, the presence of direct repeats at their ends, the association of pathogenicity islands with transfer RNA genes, the presence of integrase determinants and other mobility loci, and their genetic instability argue for the generation of pathogenicity islands by horizontal gene transfer, a process that is well known to contribute to microbial evolution. In this article we review these and other aspects of pathogenicity islands and discuss the concept that they represent a subclass of genomic islands. Genomic islands are present in the majority of genomes of pathogenic as well as nonpathogenic bacteria and may encode accessory functions which have been previously spread among bacterial populations. PMID- 11018141 TI - DNA segregation in bacteria. AB - Segregation of DNA in bacterial cells is an efficient process that assures that every daughter cell receives a copy of genomic and plasmid DNA. In this review, we focus primarily on observations in recent years, including the visualization of DNA and proteins at the subcellular level, that have begun to define the events that separate DNA molecules. Unlike the process of chromosome segregation in higher cells, segregation of the bacterial chromosome is a continuous process in which chromosomes are separated as they are replicated. Essential to separation is the initial movement of sister origins to opposite ends of the cell. Subsequent replication and controlled condensation of DNA are the driving forces that move sister chromosomes toward their respective origins, which establishes the polarity required for segregation. Final steps in the resolution and separation of sister chromosomes occur at the replication terminus, which is localized at the cell center. In contrast to the chromosome, segregation of low copy plasmids, such as Escherichia coli F, P1, and R1, is by mechanisms that resemble those used in eukaryotic cells. Each plasmid has a centromere-like site to which plasmid-specified partition proteins bind to promote segregation. Replication of plasmid DNA, which occurs at the cell center, is followed by rapid partition protein-mediated separation of sister plasmids, which become localized at distinct sites on either side of the division plane. The fundamental similarity between chromosome and plasmid segregation-placement of DNA to specific cell sites-implies an underlying cellular architecture to which both DNA and proteins refer. PMID- 11018142 TI - Polyphosphate and phosphate pump. AB - In microbial cells, inorganic polyphosphate (polyP) plays a significant role in increasing cell resistance to unfavorable environmental conditions and in regulating different biochemical processes. polyP is a polyfunctional compound. The most important of its functions are the following: phosphate and energy reservation, cation sequestration and storage, membrane channel formation, participation in phosphate transport, involvement in cell envelope formation and function, gene activity control, regulation of enzyme activities, and a vital role in stress response and stationary-phase adaptation. The functions of polyP have changed greatly during the evolution of living organisms. In prokaryotes, the most important functions are as an energy source and a phosphate reserve. In eukaryotic microorganisms, the regulatory functions predominate. Therefore, a great difference is observed between prokaryotes and eukaryotes in their polyP metabolizing enzymes. Some key prokaryotic enzymes are not present in eukaryotes, and conversely, eukaryotes have developed new polyP-metabolizing enzymes that are not present in prokaryotes. The synthesis and degradation of polyP in each specialized organelle and compartment of eukaryotic cells are mediated by different sets of enzymes. This is consistent with the endosymbiotic hypothesis of eukaryotic cell origin. PMID- 11018143 TI - Assembly and function of type III secretory systems. AB - Type III secretion systems allow Yersinia spp., Salmonella spp., Shigella spp., Bordetella spp., and Pseudomonas aeruginosa and enteropathogenic Escherichia coli adhering at the surface of a eukaryotic cell to inject bacterial proteins across the two bacterial membranes and the eukaryotic cell membrane to destroy or subvert the target cell. These systems consist of a secretion apparatus, made of approximately 25 proteins, and an array of proteins released by this apparatus. Some of these released proteins are "effectors," which are delivered into the cytosol of the target cell, whereas the others are "translocators," which help the effectors to cross the membrane of the eukaryotic cell. Most of the effectors act on the cytoskeleton or on intracellular-signaling cascades. A protein injected by the enteropathogenic E. coli serves as a membrane receptor for the docking of the bacterium itself at the surface of the cell. Type III secretion systems also occur in plant pathogens where they are involved both in causing disease in susceptible hosts and in eliciting the so-called hypersensitive response in resistant or nonhost plants. They consist of 15-20 Hrp proteins building a secretion apparatus and two groups of effectors: harpins and avirulence proteins. Harpins are presumably secreted in the extracellular compartment, whereas avirulence proteins are thought to be targeted into plant cells. Although a coherent picture is clearly emerging, basic questions remain to be answered. In particular, little is known about how the type III apparatus fits together to deliver proteins in animal cells. It is even more mysterious for plant cells where a thick wall has to be crossed. In spite of these haunting questions, type III secretion appears as a fascinating trans-kingdom communication device. PMID- 11018144 TI - Proteins shared by the transcription and translation machines. AB - It is becoming increasingly clear that the complex machines involved in transcription and translation, the two major activities leading to gene expression, communicate directly with one another by sharing proteins. For some proteins, such as ribosomal proteins S10 and L4, there is strong evidence of their participation in both processes, and much is known about their role in both activities. The exact roles and interactions of other proteins, such as Nus factors B and G, in both transcription and translation remain a mystery. Although there are not, at present, many examples of such shared proteins, the importance of understanding their behavior and intimate involvement with two major cellular machines is beginning to be appreciated. Studies related to the dual activities of these proteins and searches for more examples of proteins shared between the transcription and translation machines should lead to a better understanding of the communication between these two activities and the purposes it serves. PMID- 11018145 TI - Holins: the protein clocks of bacteriophage infections. AB - Two proteins, an endolysin and a holin, are essential for host lysis by bacteriophage. Endolysin is the term for muralytic enzymes that degrade the cell wall; endolysins accumulate in the cytosol fully folded during the vegetative cycle. Holins are small membrane proteins that accumulate in the membrane until, at a specific time that is "programmed" into the holin gene, the membrane suddenly becomes permeabilized to the fully folded endolysin. Destruction of the murein and bursting of the cell are immediate sequelae. Holins control the length of the infective cycle for lytic phages and so are subject to intense evolutionary pressure to achieve lysis at an optimal time. Holins are regulated by protein inhibitors of several different kinds. Holins constitute one of the most diverse functional groups, with >100 known or putative holin sequences, which form >30 ortholog groups. PMID- 11018146 TI - Oxygen respiration by desulfovibrio species. AB - Throughout the first 90 years after their discovery, sulfate-reducing bacteria were thought to be strict anaerobes. During the last 15 years, however, it has turned out that they have manifold properties that enable them to cope with oxygen. Sulfate-reducing bacteria not only survive oxygen exposure for at least days, but many of them even reduce oxygen to water. This process can be a true respiration process when it is coupled to energy conservation. Various oxygen reducing systems are present in Desulfovibrio species. In Desulfovibrio vulgaris and Desulfovibrio desulfuricans, oxygen reduction was coupled to proton translocation and ATP conservation. In these species, the periplasmic fraction, which contains hydrogenase and cytochrome c3, was found to catalyze oxygen reduction with high rates. In Desulfovibrio gigas, a cytoplasmic rubredoxin oxidase was identified as an oxygen-reducing terminal oxidase. Generally, the same substrates as with sulfate are oxidized with oxygen. As additional electron donors, reduced sulfur compounds can be oxidized to sulfate. Sulfate-reducing bacteria are thus able to catalyze all reactions of a complete sulfur cycle. Despite a high respiration rate and energy coupling, aerobic growth of pure cultures is poor or absent. Instead, the respiration capacity appears to have a protective function. High numbers of sulfate-reducing bacteria are present in the oxic zones and near the oxic-anoxic boundaries of sediments and in stratified water bodies, microbial mats and termite guts. Community structure analyses and microbiological studies have shown that the populations in those zones are especially adapted to oxygen. How dissimilatory sulfate reduction can occur in the presence of oxygen is still enigmatic, because in pure culture oxygen blocks sulfate reduction. Behavioral responses to oxygen include aggregation, migration to anoxic zones, and aerotaxis. The latter leads to band formation in oxygen containing zones at concentrations of 40 in some cases) is directly controlled by the prevailing intracellular concentration of Fe(II) via its complexing to a regulatory protein (the Fur protein or equivalent). In this way, the biochemistry of the bacterial cell can accommodate the challenges from the host. Agents that interfere with bacterial iron metabolism may prove extremely valuable for chemotherapy of diseases. PMID- 11018149 TI - Consistency of SINE insertion topology and flanking sequence tree: quantifying relationships among cetartiodactyls. AB - Short interspersed nuclear elements (SINEs) have been used to generate unambiguous phylogenetic topologies relating eukaryotic taxa. The irreversible nature of SINE retroposition is supported by a large body of comparative genome data and is a fundamental assumption inherent in the value of this qualitative method of inference. Here, we assess the key assumption of unidirectional SINE insertion by comparing the SINE insertion-derived topology and the phylogenetic tree based on seven independent loci of five taxa in the order Cetartiodactyla (Cetacea + Artiodactyla). The data sets and analyses were largely independent, but the loci were, by definition, linked, and thus their consistency supported an irreversible pattern of SINE retroposition. Moreover, our analyses of the flanking sequences provided estimates of divergence times among cetartiodactyl lineages unavailable from SINE insertion analysis alone. Unexpected rate heterogeneity among sites of SINE-flanking sequences and other noncoding DNA sequences were observed. Sequence simulations suggest that this rate heterogeneity may be an artifact resulting from the inaccuracies of the substitution model used. PMID- 11018150 TI - MADS-Box gene diversity in seed plants 300 million years ago. AB - MADS-box genes encode a family of transcription factors which control diverse developmental processes in flowering plants ranging from root development to flower and fruit development. Through phylogeny reconstructions, most of these genes can be subdivided into defined monophyletic gene clades whose members share similar expression patterns and functions. Therefore, the establishment of the diversity of gene clades was probably an important event in land plant evolution. In order to determine when these clades originated, we isolated cDNAs of 19 different MADS-box genes from Gnetum gnemon, a gymnosperm model species and thus a representative of the sister group of the angiosperms. Phylogeny reconstructions involving all published MADS-box genes were then used to identify gene clades containing putative orthologs from both angiosperm and gymnosperm lineages. Thus, the minimal number of MADS-box genes that were already present in the last common ancestor of extant gymnosperms and angiosperms was determined. Comparative expression studies involving pairs of putatively orthologous genes revealed a diversity of patterns that has been largely conserved since the time when the angiosperm and gymnosperm lineages separated. Taken together, our data suggest that there were already at least seven different MADS-box genes present at the base of extant seed plants about 300 MYA. These genes were probably already quite diverse in terms of both sequence and function. In addition, our data demonstrate that the MADS-box gene families of extant gymnosperms and angiosperms are of similar complexities. PMID- 11018151 TI - Cirripede phylogeny using a novel approach: molecular morphometrics. AB - We present a new method using nucleic acid secondary structure to assess phylogenetic relationships among species. In this method, which we term "molecular morphometrics," the measurable structural parameters of the molecules (geometrical features, bond energies, base composition, etc.) are used as specific characters to construct a phylogenetic tree. This method relies both on traditional morphological comparison and on molecular sequence comparison. Applied to the phylogenetic analysis of Cirripedia, molecular morphometrics supports the most recent morphological analyses arguing for the monophyly of Cirripedia sensu stricto (Thoracica + Rhizocephala + Acrothoracica). As a proof, a classical multiple alignment was also performed, either using or not using the structural information to realign the sequence segments considered in the molecular morphometrics analysis. These methods yielded the same tree topology as the direct use of structural characters as a phylogenetic signal. By taking into account the secondary structure of nucleic acids, the new method allows investigators to use the regions in which multiple alignments are barely reliable because of a large number of insertions and deletions. It thus appears to be complementary to classical primary sequence analysis in phylogenetic studies. PMID- 11018152 TI - Maximum-likelihood analysis of molecular adaptation in abalone sperm lysin reveals variable selective pressures among lineages and sites. AB - Maximum-likelihood models of codon substitution were used to analyze sperm lysin genes of 25 abalone (HALIOTIS:) species to identify lineages and amino acid sites under diversifying selection. The models used the nonsynonymous/synonymous rate ratio (omega = d(N)/d(S)) as an indicator of selective pressure and allowed the ratio to vary among lineages or sites. Likelihood ratio tests suggested significant variation in selective pressure among lineages. The variable selective pressure provided an explanation for the previous observation that the omega ratio is >1 in comparisons of closely related species and <1 in comparisons of distantly related species. Computer simulations demonstrated that saturation of nonsynonymous substitutions and constraint on lysin structure were unlikely to account for the observed pattern. Lineages linking closely related sympatric species appeared to be under diversifying selection, while lineages separating distantly related species from different geographic locations were associated with low evolutionary rates. The selective pressure indicated by the omega ratio was found to vary greatly among amino acid sites in lysin. Sites under potential diversifying selection were identified. Ancestral lysins were inferred to trace the route of evolution at individual sites and to provide lysin sequences for future laboratory studies. PMID- 11018153 TI - Origin and evolution of eukaryotic chaperonins: phylogenetic evidence for ancient duplications in CCT genes. AB - Chaperonins are oligomeric protein-folding complexes which are divided into two distantly related structural classes. Group I chaperonins (called GroEL/cpn60/hsp60) are found in bacteria and eukaryotic organelles, while group II chaperonins are present in archaea and the cytoplasm of eukaryotes (called CCT/TriC). While archaea possess one to three chaperonin subunit-encoding genes, eight distinct CCT gene families (paralogs) have been characterized in eukaryotes. We are interested in determining when during eukaryotic evolution the multiple gene duplications producing the CCT subunits occurred. We describe the sequence and phylogenetic analysis of five CCT genes from TRICHOMONAS: vaginalis and seven from GIARDIA: lamblia, representatives of amitochondriate protist lineages thought to have diverged early from other eukaryotes. Our data show that the gene duplications producing the eight CCT paralogs took place prior to the organismal divergence of TRICHOMONAS: and GIARDIA: from other eukaryotes. Thus, these divergent protists likely possess completely hetero-oligomeric CCT complexes like those in yeast and mammalian cells. No close phylogenetic relationship between the archaeal chaperonins and specific CCT subunits was observed, suggesting that none of the CCT gene duplications predate the divergence of archaea and eukaryotes. The duplications producing the CCTdelta and CCTepsilon subunits, as well as CCTalpha, CCTbeta, and CCTeta, are the most recent in the CCT gene family. Our analyses show significant differences in the rates of evolution of archaeal chaperonins compared with the eukaryotic CCTs, as well as among the different CCT subunits themselves. We discuss these results in light of current views on the origin, evolution, and function of CCT complexes. PMID- 11018155 TI - Comparative evolutionary analysis of chalcone synthase and alcohol dehydrogenase loci in Arabidopsis, Arabis, and related genera (Brassicaceae). AB - We analyzed sequence variation for chalcone synthase (Chs) and alcohol dehydrogenase (Adh) loci in 28 species in the genera Arabidopsis and Arabis and related taxa from tribe Arabideae. Chs was single-copy in nearly all taxa examined, while Adh duplications were found in several species. Phylogenies constructed from both loci confirmed that the closest relatives of Arabidopsis thaliana include Arabidopsis lyrata, Arabidopsis petraea, and Arabidopsis halleri (formerly in the genus Cardaminopsis). Slightly more distant are the North American n = 7 Arabis (Boechera) species. The genus Arabis is polyphyletic-some unrelated species appear within this taxonomic classification, which has little phylogenetic meaning. Fossil pollen data were used to compute a synonymous substitution rate of 1.5 x 10 substitutions per site per year for both Chs and Adh. Arabidopsis thaliana diverged from its nearest relatives about 5 MYA, and from Brassica roughly 24 MYA. Independent molecular and fossil data from several sources all provide similar estimates of evolutionary timescale in the Brassicaceae. PMID- 11018154 TI - Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum. AB - Multilocus genotyping of microbial pathogens has revealed a range of population structures, with some bacteria showing extensive recombination and others showing almost complete clonality. The population structure of the protozoan parasite Plasmodium falciparum has been harder to evaluate, since most studies have used a limited number of antigen-encoding loci that are known to be under strong selection. We describe length variation at 12 microsatellite loci in 465 infections collected from 9 locations worldwide. These data reveal dramatic differences in parasite population structure in different locations. Strong linkage disequilibrium (LD) was observed in six of nine populations. Significant LD occurred in all locations with prevalence <1% and in only two of five of the populations from regions with higher transmission intensities. Where present, LD results largely from the presence of identical multilocus genotypes within populations, suggesting high levels of self-fertilization in populations with low levels of transmission. We also observed dramatic variation in diversity and geographical differentiation in different regions. Mean heterozygosities in South American countries (0.3-0.4) were less than half those observed in African locations (0. 76-0.8), with intermediate heterozygosities in the Southeast Asia/Pacific samples (0.51-0.65). Furthermore, variation was distributed among locations in South America (F:(ST) = 0.364) and within locations in Africa (F:(ST) = 0.007). The intraspecific patterns of diversity and genetic differentiation observed in P. falciparum are strikingly similar to those seen in interspecific comparisons of plants and animals with differing levels of outcrossing, suggesting that similar processes may be involved. The differences observed may also reflect the recent colonization of non-African populations from an African source, and the relative influences of epidemiology and population history are difficult to disentangle. These data reveal a range of population structures within a single pathogen species and suggest intimate links between patterns of epidemiology and genetic structure in this organism. PMID- 11018156 TI - Rapid evolution of the family of CONSTANS LIKE genes in plants. AB - A family of CONSTANS LIKE genes (COLs) has recently been identified in Arabidopsis thaliana and other plant species. CONSTANS, the first isolated member, is a putative zinc finger transcription factor that promotes the induction of flowering in A. thaliana in long photoperiods. Phylogenetic analysis of the COL family demonstrated that it is organized into a few distinct groups, some of which evolved before the divergence of gymnosperms and angiosperms. Molecular evolutionary analyses showed that COL genes within the Brassicaceae family evolve rapidly. The number of nonsynonymous substitutions was larger, and the ratio of nonsynonymous to synonymous substitutions was higher. The analysis also indicated that the rate of evolution is heterogeneous between different domains in the COL genes. The results support previous data indicating that plant regulatory genes evolve relatively fast and that the rate of evolution varies significantly between different regions of those genes. The rate of evolution of COL genes seems to have accelerated during later stages of evolution, possibly as an effect of frequent gene duplications. PMID- 11018157 TI - Cytochrome c oxidase assembly in primates is sensitive to small evolutionary variations in amino acid sequence. AB - Respiring mitochondria require many interactions between nuclear and mitochondrial genomes. Although mitochondrial DNA (mtDNA) from the gorilla and the chimpanzee are able to restore oxidative phosphorylation in a human cell, mtDNAs from more distant primate species are functionally incompatible with human nuclear genes. Using microcell-mediated chromosome and mitochondria transfer, we introduced and maintained a functional orangutan mtDNA in a human nuclear background. However, partial oxidative phosphorylation function was restored only in the presence of most orangutan chromosomes, suggesting that human oxidative phosphorylation-related nuclear-coded genes are not able to replace many orangutan ones. The respiratory capacity of these hybrids was decreased by 65% 80%, and cytochrome c oxidase (COX) activity was decreased by 85%-95%. The function of other respiratory complexes was not significantly altered. The translation of mtDNA-coded COX subunits was normal, but their steady-state levels were approximately 10% of normal ones. Nuclear-coded COX subunits were loosely associated with mitochondrial membranes, a characteristic of COX assembly defective mutants. Our results suggest that many human nuclear-coded genes not only cannot replace the orangutan counterparts, but also exert a specific interference at the level of COX assembly. This cellular model underscores the precision of COX assembly in mammals and sheds light on the nature of nuclear mtDNA coevolutionary constraints. PMID- 11018158 TI - An amphioxus Emx homeobox gene reveals duplication during vertebrate evolution. AB - Members of the Emx homeobox gene class are expressed during embryogenesis in the brain and/or other head structures of phylogenetically diverse phyla. Here, we describe sequence, genomic structure, and molecular phylogenetic analysis of a cephalochordate (amphioxus) Emx class gene termed AmphiEmxA. The genomic structure of AmphiEmxA is very similar to that of vertebrate Emx genes, with two conserved intron sites. The Drosophila homolog empty spiracles (ems) has just one intron, which may be shared with chordates; the other has been secondarily lost in this Drosophila gene and in a cnidarian Emx-related gene. We identify a highly conserved peptide motif close to the amino terminus of Emx proteins, demonstrate its similarity to a sequence found in a variety of transcription factors, and argue that it arose through convergent evolution in homeobox and forkhead genes. Finally, our molecular phylogenetic analysis strongly supports the presence of a single Emx gene in the ancestor of chordates and gene duplication along the vertebrate lineage. PMID- 11018159 TI - Multiple maxima of likelihood in phylogenetic trees: an analytic approach. AB - Maximum likelihood (ML) is a widely used criterion for selecting optimal evolutionary trees. However, the nature of the likelihood surface for trees is still not sufficiently understood, especially with regard to the frequency of multiple optima. Here, we initiate an analytic study for identifying sequences that generate multiple optima. We concentrate on the problem of optimizing edge weights for a given tree or trees (as opposed to searching through the space of all trees). We report a new approach to computing ML directly, which we have used to find large families of sequences that have multiple optima, including sequences with a continuum of optimal points. Such data sets are best supported by different (two or more) phylogenies that vary significantly in their timings of evolutionary events. Some standard biological processes can lead to data with multiple optima, and consequently the field needs further investigation. Our results imply that hill-climbing techniques as currently implemented in various software packages cannot guarantee that one will find the global ML point, even if it is unique. PMID- 11018160 TI - Horizontal transfer and selection in the evolution of P elements. AB - The roles of selection and horizontal transfer in the evolution of the canonical subfamily of P: elements were studied in the saltans and willistoni species groups of the genus Drosophila (subgenus Sophophora). We estimate that the common ancestor of the canonical P: subfamily dates back 2-3 Myr at the most, despite the much older age (more than 40 Myr) of the P: family as a whole. The evolution of the canonical P: subfamily is characterized by weak selection at nonsynonymous sites. These sites have evolved at three quarters the rate of synonymous sites, in which no selective constraints were detected. Their recent horizontal transfer best explains the high degree of similarity among canonical P: elements from the saltans and willistoni species groups. These results are consistent with a model of P:-element evolution in which selective constraints are imposed at the time of horizontal transfer. Furthermore, it is estimated that the spread and diversification of the canonical subfamily involved a minimum of 11 horizontal transfer events among the 18 species surveyed within the past 3 Myr. The presence of multiple P: subfamilies in the saltans and willistoni species groups is likely to be the result of multiple invasions that have previously swept through these taxa in a succession of horizontal transfer events. These results suggest that horizontal transfer among eukaryotes might be more common than anticipated. PMID- 11018161 TI - Evolutionary diversification of protein-coding genes of hantaviruses. AB - Phylogenetic analyses of the S:, M, and L: genes of the hantaviruses (Bunyaviridae: Hantavirus) revealed three well-differentiated clades corresponding to viruses parasitic on three subfamilies (Murinae, Arvicolinae, and Sigmodontinae) of the rodent family Muridae. In rooted trees of M: and L: genes, the viruses with hosts belonging to Murinae formed an outgroup to those with hosts in Arvicolinae and Sigmodontinae. This phylogeny corresponded with a phylogeny of the murid subfamilies based on mitochondrial cytochrome b sequences, supporting the hypothesis that hantaviruses have coevolved with their mammalian hosts at least since the common ancestor of these three subfamilies, which probably occurred about 50 MYA. The nucleocapsid protein (encoded by the S: gene) differentiated among the viruses parasitic on the three subfamilies in such a way that a high frequency of amino acid residue charge changes occurred in a hypervariable (HV) portion of the molecule, and nonsynonymous nucleotide differences causing amino acid charge changes in the HV region occurred significantly more frequently than expected under random substitution. Along with evidence that at least in some hantaviruses the HV region is a target for host antibodies and the known importance of charged residues in determining antibody epitopes, these results suggest that changes in the HV region may represent adaptation to host-specific characteristics of the immune response. PMID- 11018162 TI - c-mos variation in songbirds: molecular evolution, phylogenetic implications, and comparisons with mitochondrial differentiation. AB - Nucleotide sequences from the c-mos proto-oncogene have previously been used to reconstruct the phylogenetic relationships between distantly related vertebrate taxa. To explore c-mos variation at shallower levels of avian divergence, we compared c-mos sequences from representative passerine taxa that span a range of evolutionary differentiation, from basal passerine lineages to closely allied genera. Phylogenetic reconstructions based on these c-mos sequences recovered topologies congruent with previous DNA-DNA hybridization-based reconstructions, with many nodes receiving high support, as indicated by bootstrap and reliability values. One exception was the relationship of Acanthisitta to the remaining passerines, where the c-mos-based searches indicated a three-way polytomy involving the Acanthisitta lineage and the suboscine and oscine passerine clades. We also compared levels of c-mos and mitochondrial differentiation across eight oscine passerine taxa and found that c-mos nucleotide substitutions accumulate at a rate similar to that of transversion substitutions in mitochondrial protein coding genes. These comparisons suggest that nuclear-encoded loci such as c-mos provide a temporal window of phylogenetic resolution that overlaps the temporal range where mitochondrial protein-coding sequences have their greatest utility and that c-mos substitutions and mtDNA transversions can serve as complementary, informative, and independent phylogenetic markers for the study of avian relationships. PMID- 11018163 TI - Recombination and the molecular clock. PMID- 11018164 TI - A trial of shortened zidovudine regimens to prevent mother-to-child transmission of human immunodeficiency virus type 1. Perinatal HIV Prevention Trial (Thailand) Investigators. AB - BACKGROUND: The optimal duration of zidovudine administration to prevent perinatal transmission of human immunodeficiency virus type 1 (HIV-1) should be determined to facilitate its use in areas where resources are limited. METHODS: We conducted a randomized, double-blind equivalence trial of zidovudine starting in the mother at 28 weeks' gestation, with 6 weeks of treatment in the infant (the long-long regimen), which is similar to protocol 076; zidovudine starting at 35 weeks' gestation, with 3 days of treatment in the infant (the short-short regimen); a long-short regimen; and a short-long regimen. The mothers received zidovudine orally during labor. The infants were fed formula and were tested for HIV DNA at 1, 45, 120, and 180 days. After the first interim analysis, the short short regimen was stopped. RESULTS: A total of 1437 women were enrolled. At the first interim analysis, the rates of HIV transmission were 4.1 percent for the long-long regimen and 10.5 percent for the short-short regimen (P=0.004). For the entire study period, the transmission rates were 6.5 percent (95 percent confidence interval, 4.1 to 8.9 percent) for the long-long regimen, 4.7 percent (95 percent confidence interval, 2.4 to 7.0 percent) for the long-short regimen, and 8.6 percent (95 percent confidence interval, 5.6 to 11.6 percent) for the short-long regimen. The rate of in utero transmission was significantly higher with the two regimens with shorter maternal treatment (5.1 percent) than with the two with longer maternal treatment (1.6 percent). CONCLUSIONS: The short-short zidovudine regimen is inferior to the long-long regimen and leads to a higher rate of perinatal HIV transmission. The long-short, short-long, and long-long regimens had equivalent efficacy. However, the higher rate of in utero transmission with the short-long regimen suggests that longer treatment of the infant cannot substitute for longer treatment of the mother. PMID- 11018165 TI - A prospective study of asymptomatic bacteriuria in sexually active young women. AB - BACKGROUND: Asymptomatic bacteriuria is common in young women, but little is known about its pathogenesis, natural history, risk factors, and temporal association with symptomatic urinary tract infection. METHODS: We prospectively evaluated 796 sexually active, nonpregnant women from 18 through 40 years of age over a period of six months for the occurrence of asymptomatic bacteriuria (defined as at least 10(5) colony-forming units of urinary tract pathogens per milliliter). The women were patients at either a university student health center or a health maintenance organization. Periodic urine cultures were taken, daily diaries were kept, and regularly scheduled interviews were performed. Escherichia coli strains were tested for hemolysin, the papG genotype, and the ribosomal RNA type. RESULTS: The prevalence of asymptomatic bacteriuria (the proportion of urine cultures with bacteriuria in asymptomatic women) was 5 percent (95 percent confidence interval, 4 percent to 6 percent) among women in the university group and 6 percent (95 percent confidence interval, 5 percent to 8 percent) among women in the health-maintenance-organization group. Persistent asymptomatic bacteriuria with the same E. coli strain was rare. Symptomatic urinary tract infection developed within one week after 8 percent of occasions on which a culture showed asymptomatic bacteriuria, as compared with 1 percent of occasions when asymptomatic bacteriuria was not found (P<0.001). Asymptomatic bacteriuria was associated with the same risk factors as for symptomatic urinary tract infection, particularly the use of a diaphragm plus spermicide and sexual intercourse. CONCLUSIONS: Asymptomatic bacteriuria in young women is common but rarely persists. It is a strong predictor of subsequent symptomatic urinary tract infection. PMID- 11018166 TI - Central diabetes insipidus in children and young adults. AB - BACKGROUND: Central diabetes insipidus is rare in children and young adults, and up to 50 percent of cases are idiopathic. The clinical presentation and the long term course of this disorder are largely undefined. METHODS: We studied all 79 patients with central diabetes insipidus who were seen at four pediatric endocrinology units between 1970 and 1996. There were 37 male and 42 female patients whose median age at diagnosis was 7.0 years (range, 0.1 to 24.8). All patients underwent magnetic resonance imaging (MRI) and periodic studies of anterior pituitary function. The median duration of follow-up was 7.6 years (range, 1.6 to 26.2). RESULTS: The causes of the central diabetes insipidus were Langerhans-cell histiocytosis in 12 patients, an intracranial tumor in 18 patients, a skull fracture in 2 patients, and autoimmune polyendocrinopathy in 1 patient; 5 patients had familial disease. The cause was considered to be idiopathic in 41 patients (52 percent). In 74 patients (94 percent) the posterior pituitary was not hyperintense on the first MRI scan obtained, and 29 patients (37 percent) had thickening of the pituitary stalk. Eighteen patients had changes in the thickness of the pituitary stalk over time, ranging from normalization (six patients) or a decrease in thickness (one patient) to further thickening (seven patients) or thickening of a previously normal stalk (four patients). Anterior pituitary hormone deficiencies, primarily growth hormone deficiency, were documented in 48 patients (61 percent) a median of 0.6 year (range, 0.1 to 18.0) after the onset of central diabetes insipidus. CONCLUSIONS: Most children and young adults with acquired central diabetes insipidus have abnormal findings on MRI scans of the head, which may change over time, and at least half have anterior pituitary hormone deficiencies during follow-up. PMID- 11018167 TI - The neuropathic postural tachycardia syndrome. AB - BACKGROUND: The postural tachycardia syndrome is a common disorder that is characterized by chronic orthostatic symptoms and a dramatic increase in heart rate on standing, but that does not involve orthostatic hypotension. Several lines of evidence indicate that this disorder may result from sympathetic denervation of the legs. METHODS: We measured norepinephrine spillover (the rate of entry of norepinephrine into the venous circulation) in the arms and legs both before and in response to exposure to three stimuli (the cold pressor test, sodium nitroprusside infusion, and tyramine infusion) in 10 patients with the postural tachycardia syndrome and in 8 age- and sex-matched normal subjects. RESULTS: At base line, the mean (+/-SD) plasma norepinephrine concentration in the femoral vein was lower in the patients with the postural tachycardia syndrome than in the normal subjects (135+/-30 vs. 215+/-55 pg per milliliter [0.80+/-0.18 vs. 1.27+/-0.32 nmol per liter], P=0.001). Norepinephrine spillover in the arms increased to a similar extent in the two groups in response to each of the three stimuli, but the increases in the legs were smaller in the patients with the postural tachycardia syndrome than in the normal subjects (0.001+/-0.09 vs. 0.12+/-0.12 ng per minute per deciliter of tissue [0.006+/-0.53 vs. 0.71+/-0.71 nmol per minute per deciliter] with the cold pressor test, P=0.02; 0.02+/-0.07 vs. 0.23+/-0.17 ng per minute per deciliter [0.12+/-0.41 vs. 1.36+/-1.00 nmol per minute per deciliter] with nitroprusside infusion, P=0.01; and 0.008+/-0.09 vs. 0.19+/-0.25 ng per minute per deciliter [0.05+/-0.53 vs. 1.12+/-1.47 nmol per minute per deciliter] with tyramine infusion, P=0.04). CONCLUSIONS: The neuropathic postural tachycardia syndrome results from partial sympathetic denervation, especially in the legs. PMID- 11018168 TI - Mutations in coagulation factors in women with unexplained late fetal loss. AB - BACKGROUND: Factor V and prothrombin-gene mutations are independent risk factors for venous thrombosis; it is debated whether a mutation in the gene encoding methylenetetrahydrofolate reductase, an enzyme involved in homocysteine metabolism, also increases the risk of venous thrombosis. Whether any of these mutations is associated with an increased risk of late fetal death is not known. METHODS: We studied 67 women with a first episode of unexplained late fetal loss (fetal death after 20 weeks or more of gestation) and 232 women who had had one or more normal pregnancies and no late fetal losses. All the women were tested for the presence of three gene mutations. Women with other thrombophilic conditions were excluded from the study. RESULTS: Eleven of the 67 women with late fetal loss (16 percent) and 13 of the 232 control women (6 percent) had either the factor V or the prothrombin mutation. The relative risks of late fetal loss in carriers of the factor V and prothrombin mutations were 3.2 (95 percent confidence interval, 1.0 to 10.9) and 3.3 (95 percent confidence interval, 1.1 to 10.3), respectively. Thirteen percent of the women whose fetuses died and 20 percent of the control women were homozygous for the mutation in the methylenetetrahydrofolate reductase gene (relative risk, 0.8; 95 percent confidence interval, 0.5 to 1.2). CONCLUSIONS: Both the factor V and the prothrombin mutations are associated with an approximate tripling of the risk of late fetal loss. PMID- 11018169 TI - A Medical Mystery. PMID- 11018170 TI - T-cell function and migration. Two sides of the same coin. PMID- 11018171 TI - Preventing vertical transmission of HIV infection. PMID- 11018172 TI - Asymptomatic bacteriuria--important or not? PMID- 11018188 TI - Effects of tirofiban plus heparin versus heparin alone on troponin I levels in patients with acute coronary syndromes. AB - Elevated serum troponins following an acute coronary syndrome (ACS) predict a poor clinical outcome. Glycoprotein (GP) IIb/IIIa inhibitors reduce adverse clinical outcomes in patients with ACS, although their effect on serum troponin I (TnI) in this setting has not been described. We therefore studied the effects of the GP IIb/IIIa inhibitor tirofiban on serum TnI levels in a group of patients in the Platelet Inhibition in Ischemic Syndrome Management in Patients Limited by Unstable Signs and Symptoms trial. Serial blood samples were obtained in 53 patients receiving the combination therapy of tirofiban/heparin and in 52 receiving heparin alone, and were analyzed for baseline, peak, and mean concentrations of TnI. Baseline TnI levels were not different between the combination therapy and heparin-only groups (1.6 +/- 3.0 vs 3.1 +/- 6.7 ng/ml, p = 0.15). The peak TnI level was significantly lower in the combination therapy group than in the heparin group (5.2 +/- 8.3 vs 15.5 +/- 29.1 ng/ml, p = 0.017), and mean levels over the initial 24-hour period were also significantly lower in the combination therapy group (3.2 +/- 5.0 vs 8.5 +/- 14.8 ng/ml, p = 0.016). In univariate analysis, combination therapy was associated with lower TnI levels, whereas in a multivariate model, the lower peak and mean TnI levels as a consequence of tirofiban/heparin compared with heparin monotherapy remained significant (peak, p = 0.029; mean, p = 0.035). Among patients with negative TnI at baseline, treatment with the combination of tirofiban/heparin compared with heparin monotherapy still resulted in significantly lower peak (2.5 +/- 5.4 vs 14.6 +/- 32.8 ng/ml, p = 0.024) and mean (1.2 +/- 2.6 vs 6.9 +/- 15.8 ng/ml, p = 0.029) TnI levels. In patients with ACS, therapy with the combination of tirofiban and heparin (compared with heparin treatment alone) resulted in lower serum TnI levels, suggesting reduced myocardial injury. PMID- 11018189 TI - Prognostic value of plasma levels of secretory type II phospholipase A2 in patients with unstable angina pectoris. AB - Plasma levels of secretory nonpancreatic type II phospholipase A2 (sPLA2) are increased in various chronic inflammatory diseases; this increase is correlated with disease severity. sPLA2 plays a possible role in atherogenesis and is highly expressed in atheromatous plaques. Thus, this study prospectively examined whether plasma levels of sPLA2 may have a prognostic value in patients with unstable angina, which is known to have inflammatory features. Plasma levels of sPLA2 were measured in 52 patients with unstable angina, in 107 patients with stable angina, and in 96 control subjects by radioimmunoassay. sPLA2 levels were significantly higher in patients with unstable angina than in those with stable angina and in control subjects. sPLA2 remained elevated after stabilization of disease. The levels were not increased in the blood in the coronary sinus. Kaplan Meier analysis demonstrated that patients with unstable angina and with the higher sPLA2 levels had a significantly higher probability of developing clinical coronary events during a follow-up period of 2 years compared with those with the lower levels. In multivariate Cox hazard analysis, the higher levels of sPLA2 were a significant predictor of developing coronary events in patients with unstable angina, independent of other risk factors, including C-reactive protein levels, an established inflammatory predictor. In conclusion, the increase in circulating levels of sPLA2 predicts clinical coronary events independently of other risk factors in patients with unstable angina. sPLA2 levels were persistently elevated but the elevated levels may not be derived from coronary circulation. PMID- 11018190 TI - Relation among exercise-induced ventricular arrhythmias, myocardial ischemia, and viability late after acute myocardial infarction. AB - This study assesses the relation between exercise-induced ventricular arrhythmia (VA) and scintigraphic markers of myocardial ischemia and viability in patients referred for exercise stress testing late after acute myocardial infarction. We studied 171 patients (144 men, age 57 +/- 10 years) with resting wall motion abnormalities by exercise stress testing in conjunction with methoxyisobutyl isonitrile (MIBI) single-photon emission computed tomography at a mean of 4.1 years after myocardial infarction. Ischemia was defined as reversible perfusion abnormalities. Myocardial viability was considered in myocardial segments with resting wall motion abnormalities in the presence of normal perfusion, a reversible defect or a fixed defect with regional MIBI uptake > or = 50% of maximal uptake. Exercise-induced VA occurred in 46 patients (27%). Patients with VA had a higher prevalence of infarct-related artery stenosis (43 [93%] vs 93 [74%], p < 0.01), peri-infarction ischemia (32 [70%] vs 54 [43%], p < 0.005), and ischemia in > or = 2 vascular regions (20 [43%] vs 27 [22%], p < 0.01) than patients without VA. Reversible defects were detected in 39 of 97 dyssynergic segments (40%) in patients with versus 40 of 248 dyssynergic segments (16%) in patients without VA (p < 0.0001). In dyssynergic segments without reversible perfusion abnormalities, the percent resting MIBI uptake was > or = 50% in 39 of 58 segments (67%) in patients with versus 63% in 131 of 208 segments in patients without VA (p = NS). The percentage of viable segments was 80% and 69% in patients with and without VA, respectively (p < 0.05). It is concluded that patients with exercise-induced VA late after myocardial infarction have a higher prevalence of ischemia in the peri-infarction zone and in multivessel distribution. Myocardial ischemia in the dyssynergic myocardium appears to be a major mechanism underlying the occurrence of VA in these patients. PMID- 11018191 TI - Ten-year trends in the incidence, treatment, and outcome of Q-wave myocardial infarction. AB - The benefits of coronary reperfusion and antiplatelet therapy for patients with Q wave acute myocardial infarction (Q-AMI) are well established in the context of randomized, controlled trials. The use and recent impact of these and other therapies on the broader, community-wide population of patients with Q-AMI is less well established. Residents of the Worcester, Massachusetts, metropolitan area (1990 census population 437,000) hospitalized with confirmed Q-AMI in all metropolitan Worcester, Massachusetts, hospitals in 4 1-year periods between 1986 and 1997 comprised the sample of interest. We examined the rates of occurrence, use of reperfusion strategies, and hospital mortality in a cohort of 711 patients with Q-AMI treated early in the reperfusion era (1986 and 1988) in comparison to 669 patients with Q-AMI treated a decade later (1995 and 1997). The percentage of Q-AMI among all hospitalized patients with AMI decreased over the decade of reperfusion therapy: 52% in 1986 and 1988 versus 35% in 1995 and 1997 (p < 0.001). Use of reperfusion therapy for patients with Q-AMI increased from 22% to 57%, with a marked increase in the use of primary angioplasty over time (1% vs 16%). The profile of patients receiving reperfusion therapy also changed significantly over the study period. Marked increases in use of antiplatelet therapy, beta blockers, angiotensin-converting enzyme inhibitors, and decreased use of calcium channel blockers, were observed over time. The crude in-hospital case fatality rate declined from 19% (1986 and 1988) to 14% (1995 and 1997) in patients with Q-AMI. Results of a multivariable regression analysis showed lack of reperfusion therapy, older age, anterior wall AMI, and cardiogenic shock to be independent predictors of in-hospital mortality in patients with Q-AMI. Thus, the percentage of all AMI's presenting as Q-AMI, and hospital mortality after Q-AMI, has decreased significantly in the past 10 years. The decrease in mortality occurs in the setting of broader use of reperfusion and adjunctive therapy (including primary angioplasty). PMID- 11018192 TI - Noninvasive assessment of reperfusion after fibrinolytic therapy for acute myocardial infarction. AB - Assessment of reperfusion by the 12-lead electrocardiogram (ECG) or biochemical markers is limited by suboptimal sensitivity and/or specificity. Body surface mapping (BSM) improves the spatial sampling of the 12-lead ECG. Serial 12-lead ECGs and 64-lead anterior BSMs were recorded from 67 patients with acute myocardial infarction undergoing coronary angiography 90 minutes after fibrinolytic therapy. ECG-1 and BSM-1 were recorded before/shortly after therapy (median 18 minutes). ECG-2 and BSM-2 were recorded after the 90-minute angiogram (median 30 minutes). The maximum ST elevation on ECG-1 was noted and > or = 30% ST resolution on ECG-2 was taken to represent partial/complete reperfusion. Patients were randomly divided into a training set and validation set. Isointegral and isopotential ST-T variables from BSMs of training-set patients were compared with Thrombolysis In Myocardial Infarction (TIMI) trial flow using discriminant analysis to identify which variables best classified reperfusion. Reperfusion (TIMI 2/3 flow) occurred in 32 of 34 training-set patients and in 29 of 33 validation-set patients. In the training set, > or = 30% ST resolution correctly classified reperfusion with 72% sensitivity (23 of 32) and 50% specificity (1 of 2). In the validation set, > or = 30% ST resolution classified reperfusion with 59% sensitivity (17 of 29) and 50% specificity (2 of 4). In comparison, a model containing 24 BSM variables correctly classified all training set patients, and when prospectively tested in the validation-set, correctly classified 28 of 29 patients who achieved reperfusion (97% sensitivity) and all 4 patients who failed to reperfuse (p = 0.035). In conclusion, BSM is more useful than the 12-lead ECG for noninvasive assessment of reperfusion after fibrinolytic therapy for acute myocardial infarction. PMID- 11018193 TI - Pravastatin reduces restenosis two years after percutaneous transluminal coronary angioplasty (REGRESS trial). AB - The Regression Growth Evaluation Statin Study (REGRESS) is a placebo-controlled multicenter study designed to assess the effect of 2-year treatment with pravastatin on the progression and regression of angiographically documented coronary artery disease. One of the secondary end points was the occurrence of 2 year restenosis in the percutaneous transluminal coronary angioplasty (PTCA) block. We randomly assigned eligible patients to receive pravastatin 40 mg once daily or placebo. The end point was the percent diameter stenosis of the target lesion at 24 months, as assessed by (semi)quantitative coronary angiography. Two hundred twenty-one patients underwent scheduled PTCA, which was considered successful in 201 patients. One hundred seventy-eight patients underwent angiographic restudy (89%). The patients in the pravastatin group (n = 109) and placebo group (n = 112) were similar at baseline. Percent diameter stenosis before angioplasty was 78 +/- 14% (mean +/- SD) in the pravastatin group and 80 +/- 14% in the placebo group (p = 0.46). At follow-up, the percent diameter stenosis was 32 +/- 23% in the pravastatin group and 45 +/- 29% in the placebo group (p < 0.001). Clinical restenosis was significantly lower in the pravastatin group (7%) compared with the placebo group (29%) (p < 0.001). Risk reduction for all events was 58%. We conclude that treatment with pravastatin reduces 2-year clinical and angiographic restenosis. PMID- 11018194 TI - Costs of revascularization over eight years in the randomized and eligible patients in the Emory Angioplasty versus Surgery Trial (EAST). AB - The Emory Angioplasty versus Surgery Trial (EAST) was a randomized trial that compared, by intention to treat, the clinical outcome and costs of percutaneous transluminal coronary angioplasty (PTCA) and coronary bypass grafting (CABG) for multivessel coronary artery disease. We present the findings of the economic analysis of EAST through 8 years of follow-up and compare the cost and outcomes of patients randomized in EAST versus patients eligible but not randomized (registry patients). Charges were assessed from hospital UB82 and UB92 bills and professional charges from the Emory Clinic. Hospital charges were reduced to cost through step-down accounting methods. All costs and charges were inflated to 1997 dollars. Costs were assessed for initial hospitalization and for cumulative costs of the initial hospitalization and additional revascularization procedures up to 8 years. Total 8-year costs were $46,548 for CABG and $44,491 for PTCA (p = 0.37). Cost of CABG in the eligible registry group showed a pattern similar to that for randomized patients, but total cost of PTCA was lower for registry patients than for randomized patients. Thus, the primary procedural costs of CABG are more than those for PTCA; this cost advantage, given the limits of measurement, is largely or even completely lost for randomized patients over the course of 8 years because of additional procedures after a first revascularization by PTCA. PMID- 11018195 TI - Relation between vascular morphologic changes during stent implantation and the magnitude of in-stent neointimal hyperplasia. AB - Intimal hyperplasia usually occurs after balloon overstretch injury or wire coil stimuli to coronary arteries. We examined whether the degree of vessel wall stretch during coronary stent placement could predict the amount of in-stent neointimal hyperplasia after a 6-month follow-up. Serial (preintervention, postballooning, poststent implantation, and a follow-up after 6 months) intravascular ultrasound (IVUS) was used to study 457 consecutive cross-sectional areas in 28 patients. IVUS imaging, using a motorized pullback system at 0.5 mm/s, allowed 1-mm axial increment measurements of the total vascular, stent, and lumen cross-sectional areas. The mean total vascular area changed from 10.89 +/- 2.50 mm2 before to 11.27 +/- 2.49 mm2 after ballooning, to 12.80 +/- 2.59 mm2 after stenting, and to 12.58 +/- 2.41 mm2 at follow-up (p < 0.0001). The mean lumen area changed from 3.36 +/- 1.95 mm2 before to 4.21 +/- 1.65 mm2 after ballooning, to 5.16 +/- 1.09 mm2 after stenting, and to 3.57 +/- 1.23 mm2 at follow-up (p < 0.0001). The mean stent area decreased from 5.25 +/- 1.17 mm2 after stenting to 5.09 +/- 0.90 mm2 at follow-up (p < 0.0001). Stepwise logistic regression analysis showed that delta total vascular area (after stent implantation - before intervention) was a strong predictor of the amount of intimal hyperplasia (r = 0.57, p < 0.0001). Vascular overstretch caused by the stenting procedure promotes intimal hyperplasia in proportion to the degree of sectional vascular stretch. PMID- 11018196 TI - Efficacy and safety of an extended-release formulation of fluvastatin for once daily treatment of primary hypercholesterolemia. AB - An extended-release (ER) formulation of fluvastatin 80 mg has been developed for once-daily treatment of primary hypercholesterolemia in patients who require fluvastatin dosages of > 40 mg/day. The study aimed to determine the efficacy and safety of the new formulation and to assess the dose response over the range of 40 to 160 mg/day. After a 4-week placebo/dietary run-in period, 123 patients with primary hypercholesterolemia (Fredrickson type IIa/IIb) were randomized to receive fluvastatin 40, 80, or 160 mg/day for 6 weeks. The 40 mg/day dosage was administered as the marketed immediate-release (IR) capsule and the 80 mg/day dosage as 1 80-mg ER tablet. Patients receiving 160 mg/day were administered 80 mg/day (1 ER tablet) for the first 2 weeks, followed by 160 mg/day (2 ER tablets) for the remainder of the study. All doses were administered once daily at bedtime. The results showed a linear dose-response relation. Doubling the fluvastatin dosage resulted in a 6% greater mean percent reduction in low-density lipoprotein cholesterol (40 mg IR -29%; 80 mg ER -35%; 160 mg ER -41%). In the 160-mg ER group, 62% of patients achieved > or = 40% reductions in low-density lipoprotein cholesterol compared with 32% and 10% of patients in the 80-mg ER and 40-mg IR groups, respectively. Dose ordering of the response was also observed for the other lipid parameters. Fluvastatin ER was well tolerated. Thus, the new ER formulation of fluvastatin was effective and well tolerated in the once-daily treatment of primary hypercholesterolemia. PMID- 11018197 TI - Role of gender and personality on quality-of-life impairment in intermittent atrial fibrillation. AB - Patients with atrial fibrillation (AF) report impaired health-related quality of life (QOL). Differences between men and women with AF have not been described and personality attributes such as somatization (tendency to amplify benign bodily sensations) may mediate potential gender differences in QOL. Patients with AF (n = 264, 59% men) who participated in the Canadian Trial of Atrial Fibrillation (n = 403) completed validated QOL questionnaires at baseline, 3 months, and 12 months after antiarrhythmic drug treatment. Women were significantly older than men and a greater proportion had hypertension, but other cardiac variables did not differ between women and men. At baseline, after controlling for significant clinical and demographic factors, women reported worse physical health (p = 0.002) and functional capacity (p < 0.001), but not mental health or general well being. Women also had more frequent and severe cardiac symptoms than men (both p < 0.001). Physical health improved significantly from baseline to 3 months for women (p = 0.002), but not for men (p = 0.066). Conversely, mental health improved for men (p = 0.007), but not for women. Cardiac symptom frequency and severity improved over time for women and men (all p < 0.001). Tendency to somatize predicted poor QOL, and women had higher scores than men (p = 0.023). However, after controlling for somatization, women still had worse physical function, functional capacity, and symptom burden than men. Independent of cardiac disease severity and age, women with AF had significantly more impaired QOL than men, specifically on domains related to physical rather than emotional functioning. Personality attributes may have a role in influencing QOL outcomes. PMID- 11018198 TI - Left atrial appendage flow velocities in subjects with normal left ventricular function. AB - The objectives of this study were to establish reference values and define the determinants of left atrial appendage (LAA) flow velocities in the general population. LAA flow velocities (contraction and filling velocities) were assessed by transesophageal echocardiography in 310 subjects aged > or = 45 years, sampled from the population-based Stroke Prevention: Assessment of Risk in a Community study. All subjects were in sinus rhythm, with preserved left ventricular systolic function (ejection fraction > or = 50%), and without valvular disease. Values of LAA contraction and filling velocities were established for various age groups in the population. Age was negatively associated with LAA contraction and filling velocities, which decreased by 4.1 cm/s (p < 0.001) and 2.0 cm/s (p < 0.01) for every 10 years of age, respectively. Contraction velocities were 5 cm/s higher in men than in women (p < 0.05). After adjusting for age and sex, heart rate was independently associated with LAA contraction velocities (p < 0.001; nonlinear association). Body surface area, left atrial size, left ventricular mass index, and a history of previous cardiac disease or hypertension showed no significant association with LAA flow velocities (p > 0.05). Furthermore, detailed analysis of 24-hour ambulatory blood pressure data (available in 253 subjects) showed no association between various blood pressure parameters (systolic and diastolic blood pressure, out-of-bed and in-bed measurements) and LAA flow velocities (all p > 0.05). In summary, the present study establishes the reference values for LAA flow velocities in a large sample of the general population. LAA flow velocities progressively decline with age in subjects with preserved left ventricular systolic function. PMID- 11018199 TI - Effect of simvastatin on restenosis after percutaneous transluminal angioplasty of femoropopliteal arterial obstruction. AB - This retrospective observational intravascular ultrasound study evaluated whether simvastatin therapy limits lumen area reduction 1-year after percutaneous transluminal angioplasty (PTA) by reducing reactive plaque growth, reducing reactive vasoconstriction, or both. This study showed that plaque growth is a general response 1 year after PTA regardless of the use of simvastatin; simvastatin has the potential to induce positive vascular remodeling, thereby reducing the occurrence of restenosis. PMID- 11018200 TI - Acute and long-term results of treatment of diffuse in-stent restenosis in aortocoronary saphenous vein grafts. AB - Treatment of diffuse in-stent restenosis in saphenous vein grafts with excimer laser coronary angioplasty plus adjunct balloon angioplasty achieves an adequate acute result. However, this population has high long-term mortality and frequent need for repeat revascularization. PMID- 11018201 TI - Safety of femoral closure devices after percutaneous coronary interventions in the era of glycoprotein IIb/IIIa platelet blockade. AB - We compared in-hospital femoral complications of Angio-Seal, Perclose, and manual compression in consecutive patients who underwent percutaneous coronary interventions in the era of glycoprotein IIb/IIIa platelet inhibition. Femoral closure devices have a similar overall risk profile as manual compression, even in patients treated with glycoprotein IIb/IIIa platelet inhibition, although certain rare complications such as retroperitoneal hemorrhage and severe access site infection may be more common with the use of these devices. PMID- 11018202 TI - Certain cardiac risk factors predict risk factor interventions and influence communication between physicians and patients. AB - We assessed predictors of cardiac risk factor testing and services and the degree of concordance among patients, physicians, and the medical records for these services, and found considerable variability among different risk factors. The data suggest that baseline risk factors influence communication and performance of interventions and that physicians appear to be underestimating the importance of treating multiple risk factors simultaneously. PMID- 11018203 TI - Prognostic usefulness of dobutamine-induced ST-segment elevation and T-wave normalization after uncomplicated acute myocardial infarction. AB - We followed 229 consecutive patients exhibiting negative T waves on infarct related electrocardiographic leads; these patients underwent dobutamine stress echocardiography within 10 days after a first uncomplicated acute myocardial infarction. T-wave normalization, but not ST-segment elevation, recognized patients at higher risk of cardiac events and optimized the prognostic accuracy of both myocardial viability and ischemia, to which it was correlated and became an independent predictor in cases of subdiagnostic stress echocardiography. PMID- 11018204 TI - In vivo uptake of azithromycin in human coronary plaques. AB - Ten patients with symptomatic coronary artery disease received oral azithromycin for 3 days and underwent directional atherectomy on the third day. Azithromycin was found in all plaque samples with a median concentration of 284 ng/ml (95% confidence interval 163 to 517 ng/ml). PMID- 11018205 TI - Long-term outcomes of minor dissection at the edge of stents detected with intravascular ultrasound. AB - We evaluated the influences of minor edge dissections on late angiographic in stent restenosis in 327 patients with 348 lesions (281 lesions without edge dissection and 67 lesions [19.3%] with edge dissection); the angiographic restenosis rate was 29.9% in the lesions with edge dissections versus 25.3% without edge dissections (p = 0.540). The minor non-flow-limiting dissections at the edge of stents may not be associated with the development of late angiographic in-stent restenosis. PMID- 11018206 TI - Effect of degree of blood pressure on the hypercoagulable state in chronic atrial fibrillation. AB - To investigate the hypothesis that higher levels of blood pressure would further promote the hypercoagulable state in atrial fibrillation (AF) by increasing the degree of hemostatic abnormalities, we studied 82 consecutive patients with chronic nonvalvular AF who were not taking antithrombotic therapy. Patients with AF had higher levels of plasma fibrin D-dimer, von Willebrand factor, and fibrinogen than controls in sinus rhythm, and no differences between tertiles of either systolic, diastolic, or mean blood pressure were found, suggesting other mechanisms may be contributing to the hypercoagulable state in AF. PMID- 11018207 TI - Microalbuminuria is associated with abnormal thoracic aortic mechanics in essential hypertension. AB - In a population of 162 patients with currently untreated essential hypertension, those with microalbuminuria (n = 75) had significantly impaired elastic properties of the proximal ascending thoracic aorta compared with their normoalbuminuric counterparts (n = 87), whereas urinary albumin excretion was a significant predictor of aortic mechanics in the entire population. Impaired aortic mechanics in microalbuminuric hypertensives were not fully accounted for by clustering of classic risk factors for atherosclerosis, and constitute a finding that may entail additional long-term cardiovascular risk in this subgroup of patients. PMID- 11018208 TI - Clinical predictors of prognosis in severe aortic stenosis in unoperated patients > or = 75 years of age. AB - In elderly patients with severe aortic stenosis, clinical evaluation can dictate decision making. Asymptomatic patients in normal sinus rhythm, without left atrial enlargement and without bundle branch block, can be safely followed clinically, regardless of echocardiographic findings. PMID- 11018209 TI - Echocardiographic predictors of survival in low gradient aortic stenosis. AB - Echocardiographic predictors of long-term survival for patients with low gradient aortic stenosis who undergo aortic valve replacement have not been previously reported. This study shows that patients with larger pre- and postoperative left ventricular volumes, a lower mean preoperative aortic pressure gradient, and failure of volumes to decrease and ejection fraction to increase postoperatively may have a poor prognosis. PMID- 11018210 TI - Diagnostic value of the electrocardiogram in suspected pulmonary embolism. AB - The electrocardiogram is shown to be of limited diagnostic value for determining pulmonary embolism in a prospective cohort study of unselected patients with suspected pulmonary embolism. PMID- 11018212 TI - Bipolar disorder: can studies of natural history help us define clinically and neurobiologically relevant subtypes? PMID- 11018213 TI - Models of bipolar disease and their clinical implications. PMID- 11018215 TI - On clinical trials in psychiatry. PMID- 11018214 TI - Treatment advances in bipolar disorder--making up for lost time. PMID- 11018216 TI - Goals for research on bipolar disorder: the view from NIMH. AB - We have much yet to accomplish in research on bipolar disorder. We must find vulnerability genes. We must identify the circuits that regulate mood, emotion, energy, and other relevant functions that are affected in bipolar disorder, and we must determine what goes wrong in those circuits during mania, depression, and other aspects of this illness. We will need to translate findings in basic neuroscience, genetics, and basic behavioral science into diverse clinical applications: novel treatments, diagnostic tools, epidemiologic approaches that could lead to preventive interventions, and surrogate markers for clinical trials. We must develop improved psychosocial interventions and test both pharmacologic and psychosocial treatments in trials that, simultaneously, improve the quality of care available and convince insurers and employers that these treatments are of substantial benefit and cost effective. The agenda is ambitious, but entirely feasible, given the scientific tools and technologies that are currently available or on the horizon. The National Institute of Mental Health is newly recommitted to harnessing these tools and technologies for the benefit of people with bipolar disorder. PMID- 11018217 TI - A consumer perspective concerning the diagnosis and treatment of bipolar disorder. PMID- 11018218 TI - Historical perspectives and natural history of bipolar disorder. AB - A review of two centuries' literature on the natural history of bipolar disorder, including modern naturalistic studies and new data from a lifelong follow-up study of 220 bipolar patients, reaches the following conclusions: the findings of modern follow-up studies are closely compatible with those of studies conducted before the introduction of modern antidepressant and mood-stabilizing treatments. Bipolar disorder has always been highly recurrent and considered to have a poor prognosis. Bipolar patients who have been hospitalized spend about 20% of their lifetime from the onset of their disorder in episodes. Fifty percent of bipolar episodes last between 2 and 7 months (median 3 months). The intervals between the first few episodes tend to shorten; later the episodes return at an irregular rhythm of about 0.4 episodes per year with high interindividual variability. Switches from mania into mild depression and from depression into hypomania were frequently reported in the 19th century and the first half of the 20th. Antidepressant and antimanic drugs have to be given as long as the natural episode lasts. Given the poor outcome of bipolar disorders found in naturalistic follow-up studies and our lifelong investigation, intensive antidepressant, antimanic, and mood-stabilizing treatments are required in most cases. Despite modern treatments the outcome into old age is still poor, full recovery without further episodes rare, recurrence of episodes with incomplete remission the rule, and the development of chronicity and suicide still frequent. PMID- 11018219 TI - Pediatric mania: a developmental subtype of bipolar disorder? AB - Despite ongoing controversy, the view that pediatric mania is rare or nonexistent has been increasingly challenged not only by case reports, but also by systematic research. This research strongly suggests that pediatric mania may not be rare but that it may be difficult to diagnose. Since children with mania are likely to become adults with bipolar disorder, the recognition and characterization of childhood-onset mania may help identify a meaningful developmental subtype of bipolar disorder worthy of further investigation. The major difficulties that complicate the diagnosis of pediatric mania include: 1) its pattern of comorbidity may be unique by adult standards, especially its overlap with attention-deficit/hyperactivity disorder, aggression, and conduct disorder; 2) its overlap with substance use disorders; 3) its association with trauma and adversity; and 4) its response to treatment is atypical by adult standards. PMID- 11018220 TI - The McLean-Harvard first-episode project: 6-month symptomatic and functional outcome in affective and nonaffective psychosis. AB - BACKGROUND: The McLean-Harvard First-Episode Project recruited affective and nonaffective patients at their first lifetime psychiatric hospitalization. METHODS: Baseline evaluation and 6-month follow-up in 257 cases yielded recovery outcomes defined by syndromal (absence of DSM-IV criteria for a current episode) and functional (vocational and residential status at least at baseline levels) status. Time to recovery was assessed by survival analysis, and risk factors by multivariate logistic regression. RESULTS: Syndromal recovery was attained by 77% of cases over an average of 84 days. By diagnostic group, syndromal recovery rates ranked (p = .001) major affective disorders (81%) > nonaffective acute psychoses (74%) > schizoaffective disorders (70%) > schizophrenia (36%). Functional recovery was significantly associated to syndromal recovery, diagnosis, shorter hospitalization normalized to year, and older age at onset. Average hospital stay declined across the study period, but recovery did not vary with year of entry. CONCLUSIONS: Syndromal recovery was achieved by nearly one half of patients within 3 months of a first lifetime hospitalization for a psychotic illness, but functional recovery was not achieved by 6 months in nearly two thirds of patients who had attained syndromal recovery. PMID- 11018221 TI - The impact of substance abuse on the course of bipolar disorder. AB - BACKGROUND: Substance abuse occurs at high rates in bipolar disorder. The reasons for this co-occurrence are unknown. Alcohol use disorders have been associated with both earlier and later age of onset of bipolar disorder, in part based on the temporal associations of the two conditions. Both drug and alcohol use disorders are associated with impaired outcome of bipolar illness. This influence may involve both direct effects of alcohol or drugs on the initiation of affective symptoms and indirect effects on treatment compliance. To extend these previous findings we examined the temporal associations of substance abuse and affective symptoms in patients with new onset bipolar disorder. METHODS: Associations between affective symptoms and alcohol and cannabis use disorder symptoms were evaluated using regression and time-series correlative methods in 50 new-onset bipolar patients. RESULTS: The duration of alcohol abuse during follow-up was associated with the time patients experienced depression. The duration of cannabis abuse was associated with the duration of mania. Several subgroups could be identified with different temporal relationships among these disorders. CONCLUSIONS: Although the relationships among substance use and bipolar disorders are complex, systematic study of the courses of the disorders might clarify how these conditions interact longitudinally. As the numbers of subjects in specific subgroups are relatively small in this study, these results should be considered preliminary. PMID- 11018222 TI - Neuropathology of bipolar disorder. AB - The literature on the neuropathology of bipolar disorder (BD) is reviewed. Postmortem findings in the areas of pathomorphology, signal transduction, neuropeptides, neurotransmitters, cell adhesion molecules, and synaptic proteins are considered. Decreased glial numbers and density in both BD and major depressive disorder (MDD) have been reported, whereas cortical neuron counts were not different in BD (in Brodmann's areas [BAs] 9 and 24). In contrast, MDD patients showed reductions in neuronal size and density (BA 9, BA 47). There are a number of findings of alterations in neuropeptides and monoamines in BD brains. Norepinephrine turnover was increased in several cortical regions and thalamus, whereas the serotonin metabolite, 5-hydroxyindoleacetic acid, and the serotonin transporter were reduced in the cortex. Several reports further implicated both cyclic adenosine monophosphate and phosphatidylinositol (PI) cascade abnormalities. G protein concentrations and activity increases were found in the occipital, prefrontal, and temporal cortices in BD. In the PI signal cascade, alterations in PKC activity were found in the prefrontal cortex. In the occipital cortex, PI hydrolysis was decreased. Two isoforms of the neural cell adhesion molecules were increased in the hippocampus of BD, whereas the synaptic protein marker, synaptophysin, was not changed. The findings of glial reduction, excess signal activity, neuropeptide abnormalities, and monoamine alterations suggest distinct imbalances in neurochemical regulation. Possible alterations in pathways involving ascending projections from the brain stem are considered. Larger numbers of BD brains are needed to further refine the conceptual models that have been proposed, and to develop coherent models of the pathophysiology of BD. PMID- 11018223 TI - Neuroimaging in bipolar disorder: what have we learned? AB - New technologies are offering increasingly powerful means to obtain structural, chemical, and functional images of the brain during life, often without the use of ionizing radiation. Bipolar disorder, with its clear physiologic features, would appear to be a prime candidate for the application of current brain imaging; however, only a modest number of studies have been reported to date, and most studies have small sample sizes and heterogeneous subject groups. Nonetheless, there are a few consistent findings among these studies, including the following: 1) Structural imaging studies suggest an increased number of white matter hyperintensities in patients with bipolar disorder. These may be lesions unique to bipolar disorder and its treatment, or related to cardiovascular risk factors, which are more common in bipolar patients. Decreased cerebellar size and anomalies of cerebellar blood volume have also been reported. Increased sulcal prominence and enlargement of the lateral and third ventricles are less consistently observed findings. 2) Spectroscopic imaging suggests abnormalities of metabolism of choline-containing compounds in symptomatically ill bipolar patients and, possibly, treatment-induced changes in choline- and myoinositol containing compounds. Each of these groups of metabolites serves as a component of membrane phospholipids and cellular second-messenger cycles. 3) Metabolic and blood flow studies provide evidence for decreased activity of the prefrontal cortex (PFC) in bipolar patients during depression. It is not clear if these changes are restricted to particular subregions of the PFC, nor if they are reversed with mania. No single pathophysiologic mechanism yet explains these findings, although all might be due to regional alterations in cellular activity and metabolism or changes in cell membrane composition and turnover. The development of imaging technologies has far outpaced their use in bipolar disorder. The promise of future studies is great, with more powerful magnetic resonance scanners, additional ligands for positron emission tomography and single photon emission computed tomography imaging, and improved image generation and processing already available. PMID- 11018224 TI - Signaling: cellular insights into the pathophysiology of bipolar disorder. AB - Clinical studies over the years have provided evidence that monoamine signaling and hypothalamic-pituitary-adrenal axis disruption are integral to the pathophysiology of bipolar disorder. A full understanding of the pathophysiology from a molecular to a systems level must await the identification of the susceptibility and protective genes driving the underlying neurobiology of bipolar disorder. Furthermore, the complexity of the unique biology of this affective disorder, which includes the predisposition to episodic and often progressive mood disturbance, and the dynamic nature of compensatory processes in the brain, coupled with limitations in experimental design, have hindered our progress to date. Imaging studies in patient populations have provided evidence of a role for anterior cingulate, amygdala, and prefrontal cortex in the pathophysiology of bipolar disorder. More recent research strategies designed to uncover the molecular mechanisms underlying our pharmacologic treatments and their interaction in the regulation of signal transduction as well as more advanced brain imaging studies remain promising approaches. This experimental strategy provides data derived from the physiologic response of the system in affected individuals and addresses the critical dynamic interaction with pharmacologic agents that effectively modify the clinical expression of the pathophysiology. PMID- 11018225 TI - Are schizophrenic and bipolar disorders related? A review of family and molecular studies. AB - Schizophrenic and bipolar disorders are similar in several epidemiologic respects, including age at onset, lifetime risk, course of illness, worldwide distribution, risk for suicide, gender influence (men and women at equal risk for both groups of disorders), and genetic susceptibility. Despite these similarities, schizophrenia and bipolar disorders are typically considered to be separate entities, with distinguishing clinical characteristics, non-overlapping etiologies, and distinct treatment regimens. Over the past three decades, multiple family studies are consistent with greater nosologic overlap than previously acknowledged. Molecular linkage studies (conducted during the 1990s) reveal that some susceptibility loci may be common to both nosologic classes. This indicates that our nosology will require substantial revision during the next decade, to reflect this shared genetic susceptibility, as specific genes are identified. PMID- 11018226 TI - Pharmacologic agents for the treatment of acute bipolar mania. AB - The knowledge base regarding the medical treatment of acute bipolar mania is rapidly expanding. Information about agents with established antimanic properties is increasing, and more agents with putative antimanic properties are being identified. We first review the controlled studies supporting the efficacy of the established antimanic agents lithium, valproate, and carbamazepine and standard antipsychotics. We then review available research on two important classes of drugs that are emerging as potential treatments for acute mania: the novel antipsychotics, which include clozapine, olanzapine, quetiapine, risperidone, and ziprasidone, and the new antiepileptics, which include gabapentin, lamotrigine, oxcarbazepine, tiagabine, topiramate, and zonisamide. We conclude that although controlled data are accumulating to support the efficacy of several atypical antipsychotics in the treatment of acute bipolar mania, particularly olanzapine, ziprasidone, and risperidone, the novel antiepileptics need more extensive study before it can be concluded that any of them possess specific antimanic properties. We also conclude that as the medical options for acute bipolar mania expand, treatment guidelines must remain both evidence based and flexible, so that they represent cutting edge medical science yet can be tailored to the specific needs of individual patients. PMID- 11018227 TI - Bipolar depression: pharmacotherapy and related therapeutic strategies. AB - The depressed phase of bipolar affective disorder is a significant cause of suffering, disability, and mortality and represents a major challenge to treating clinicians. This article first briefly reviews the phenomenology and clinical correlates of bipolar depression and then focuses on the major pharmacological treatment options. We strongly recommend use of mood stabilizers as the first line treatment for the type I form of bipolar depression, largely because longer term preventative therapy with these agents almost certainly will be indicated. Depressive episodes that do not respond to lithium, divalproex, or another mood stabilizer, or episodes that "breakthrough" despite preventive treatment, often warrant treatment with an antidepressant or electroconvulsive therapy. The necessity of mood stabilizers in the type II form of bipolar depression is less certain, aside from the rapid cycling presentation. Both experts and practicing clinicians recommend bupropion and the selective serotonin reuptake inhibitors as coequal initial choices, with venlafaxine and monoamine oxidase inhibitors, such as tranylcypromine, preferred for more resistant cases. The risk of antidepressant-induced hypomania or mania with concomitant mood stabilizer therapy is low, on the order of 5% to 10% during acute phase therapy. Additional therapeutic options and optimal durations of therapy also are discussed. PMID- 11018228 TI - Bipolar disorder therapeutics: maintenance treatment. AB - Although most of the care received by bipolar patients occurs during the maintenance phase, relatively little empirical data is available to guide long term treatment decisions. We review literature pertaining to key questions related to use of pharmacotherapy in the maintenance phase of bipolar disorder. The few double-blind trials with a reasonable sample size are restricted to bipolar I patients and address a modest range of questions mostly related to use of lithium. One rigorous multicenter trial found valproate to have prophylactic benefit. Other studies with valproate alone and in combination suggest efficacy equivalent to lithium and perhaps greater than carbamazepine. Data available for combination treatment are sparse but moderately encouraging. Maintenance treatment with standard antidepressant medications appears destabilizing for some bipolar patients, particularly following a mixed episode. Although some bipolar patients may benefit from combined treatment with a mood stabilizer and a standard antidepressant medication, current knowledge does not allow confident selection of the bipolar patients who might benefit. Clozapine and perhaps other atypical antipsychotics are promising options for maintenance treatment but have not been evaluated in double-blind trials. The numerous other agents used in maintenance treatment are primarily adjuncts to lithium, valproate, or carbamazepine, and information about them is largely anecdotal and uncontrolled. Study design for maintenance trials remains an imperfect art. Conclusions must be drawn cautiously, given the limited generalizability of study designs that accession samples enriched with presumed treatment responders, randomize patients after brief periods of partial remission, abruptly taper prior treatment, make no attempt to distinguish relapse from recurrence, use no formal outcome assessments, or report hospitalization as the only outcome criterion. PMID- 11018229 TI - Family-focused treatment of bipolar disorder: 1-year effects of a psychoeducational program in conjunction with pharmacotherapy. AB - BACKGROUND: Few studies have examined the combined effects of psychosocial treatment and pharmacotherapy for bipolar disorder. This study used a randomized, controlled design to examine a 9-month, manual-based program of family-focused psychoeducational treatment (FFT). METHODS: Bipolar patients (N = 101) were recruited shortly after an illness episode and randomly assigned to 21 sessions of FFT (n = 31) or to a comparison treatment involving two family education sessions and follow-up crisis management (CM; n = 70). Both treatments were delivered over 9 months; patients were simultaneously maintained on mood stabilizing medications. Patients were evaluated every 3 months for 1 year as to relapse status, symptom severity, and medication compliance. RESULTS: Patients assigned to FFT had fewer relapses and longer delays before relapses during the study year than did patients in CM. Patients in FFT also showed greater improvements in depressive (but not manic) symptoms. The most dramatic improvements were among FFT patients whose families were high in expressed emotion. The efficacy of FFT could not be explained by differences among patients in medication regimes or compliance. CONCLUSIONS: Family-focused psychoeducational treatment appears to be an efficacious adjunct to pharmacotherapy for bipolar disorder. Future studies should evaluate family treatment against other forms of psychotherapy matched in amount of therapist patient contact. PMID- 11018230 TI - Interpersonal and social rhythm therapy: managing the chaos of bipolar disorder. AB - Interpersonal and social rhythm therapy is an individual psychotherapy designed specifically for the treatment for bipolar disorder. Interpersonal and social rhythm therapy grew from a chronobiological model of bipolar disorder postulating that individuals with bipolar disorder have a genetic predisposition to circadian rhythm and sleep-wake cycle abnormalities that may be responsible, in part, for the symptomatic manifestations of the illness. In our model, life events (both negative and positive) may cause disruptions in patients' social rhythms that, in turn, perturb circadian rhythms and sleep-wake cycles and lead to the development of bipolar symptoms. Administered in concert with medications, interpersonal and social rhythm therapy combines the basic principles of interpersonal psychotherapy with behavioral techniques to help patients regularize their daily routines, diminish interpersonal problems, and adhere to medication regimens. It modulates both biological and psychosocial factors to mitigate patients' circadian and sleep-wake cycle vulnerabilities, improve overall functioning, and better manage the potential chaos of bipolar disorder symptomatology. PMID- 11018231 TI - Flexible treatment strategies in chronic disease: clinical and research implications. AB - Multiple treatments are available for nearly all the mood disorders. This range of treatment options adds a new dimension of choice to clinical decision making. In addition to prescribing the best initial treatment, clinicians should have an algorithm for deciding if and when to make subsequent changes in treatment to take advantage of second-line treatment options when necessary. This article aims to 1) show that a wide variety of clinical decisions can be framed as choices among adaptive (within-patient) threshold-based strategies or algorithms, illustrating the generality of the concept; 2) illustrate two ways to design randomized clinical trials to compare treatment strategies with each other to decide which strategy is best; and 3) discuss some of the advantages offered by these designs, in terms of both patient acceptability and adherence to experimental protocols. PMID- 11018232 TI - Methodological issues in developing new acute treatments for patients with bipolar illness. AB - One important aim of the recent reorganization of the National Institute of Mental Health (NIMH) is to streamline the development of new treatments for patients with severe mental illnesses, such as bipolar disorder. Researching new treatments for patients with bipolar disorder presents specific problems not readily addressed by traditional efficacy trial methodologies that aim to maximize internal validity. This article reexamines several assumptions that have guided the design of these efficacy trials but that also create obstacles for studies of bipolar disorder and suggests potential solutions. This article draws on literature from neurology and psychiatry and discussions at a MacArthur Foundation-sponsored Conference on Longitudinal Methodology in 1992 (David J. Kupfer, M.D., Chair), which brought together investigators to consider alternative designs for patients with severe and persistent mental illness. In addition, we benefited from discussions at two NIMH-sponsored conferences, one held in 1989 (Prien and Potter 1990) and the other in 1994 (Prien and Rush 1996), at which investigators and methodologists discussed issues surrounding the development and conduct of informative efficacy trials for patients with bipolar disorder. Based on these discussions and recent literature reviews, we 1) outline common problems in the development and evaluation of effective acute treatments for bipolar disorder and 2) suggest possible solutions to these impediments. We also discuss alternative designs by which to build a sequence of acute treatment studies from which efficacy, safety, and the comparative value of different treatments can be established. PMID- 11018233 TI - Changes in the structure and ultrastructure of the intestine of Spadella cephaloptera (Chaetognatha) during feeding and starvation experiments. AB - Ultrastructural changes in the intestinal epithelium of fed and starved specimens of Spadella cephaloptera are described. Animals were maintained in a circulating natural sea water system and fed with Artemia salina nauplii. After a period of acclimation, they were individually isolated, deprived of food for 24 h and submitted to controlled feeding experiments. The absorption develop in the intestinal absorptive cells (A-cells) 5 min after the ingestion of prey and consist in the formation of endocytotic vesicles and endosome-like vacuoles. During the following steps up to 10 h, a second type of digestive vacuole containing electron-dense material, and probably corresponding to a lysosome-like compartment, appears. Throughout this time, the vacuoles progressively arrange in columns, the youngest at the top and the oldest at the bottom of the A-cells. In addition, large lipid inclusions appear in the apical cytoplasm. The ultrastructural changes of the intestinal secretory cells (S-cells) is less marked, but the number of granules largely diminishes during the first 30 min after the ingestion of prey. In starved specimens, major changes in A-cells occur between the sixth and tenth day of starvation and consist in the increase of endosome-like vacuoles. Lysosome-like vacuoles containing dense material are not observed. At the same time, necrosis features are evident in S-cells. After 30 days of starvation, necrosis features are observed in the totality of the intestinal epithelium and the specimens die few days later. PMID- 11018234 TI - Avoidance of drilled gastropod shells by the hermit crab Pagurus longicarpus at Nahant, Massachusetts. AB - Most hermit crabs depend on empty gastropod shells for shelter; competition for appropriate shells is often severe. This study determined whether shells that have been drilled by naticid gastropods are suitable for occupancy by the hermit crab Pagurus longicarpus. Differences in the characteristics of empty shells and those occupied by hermit crabs were assessed at two adjacent field sites in Nahant, Massachusetts. Drilling damage was far more frequent in empty gastropod shells than in shells occupied by hermit crabs, suggesting that individuals of P. longicarpus avoid drilled shells. They did not appear to avoid shells with other forms of damage. Laboratory experiments confirmed that these hermit crabs preferentially chose intact shells over drilled shells, even when the intact shells offered were most suitable for crabs half the weight of those tested. Final shell choices were generally made within 1 h. The hermit crabs apparently discriminated between intact and drilled shells based on tactile cues, since crabs kept in the dark showed the same preference for intact shells. The hermit crabs strongly avoided, to nearly the same extent, artificially drilled shells, naturally drilled shells, and shells with holes artificially drilled on the opposite side of the shell from where they would normally be located. Possible selective forces causing P. longicarpus to show such strong behavioral avoidance of drilled shells include increased vulnerability of crabs in drilled shells to osmotic stress, predation, and eviction by conspecifics. PMID- 11018235 TI - Epibenthic amphipod abundance and predation efficiency of the pink shrimp Farfantepenaeus duorarum (Burkenroad, 1939) in habitats with different physical complexity in a tropical estuarine system. AB - Amphipod abundance and biomass were determined in soft-bottom substrates (SBS), monospecific Thalassia testudinum patches and T. testudinum with attached macroalgae (SAV) from Terminos Lagoon. Amphipods were absent in SBS, and their density and biomass were higher in SAV (3351 individualsm(-2), 1718 mg AFDWm(-2)) than in T. testudinum (1220 indm(-2), 625 mg AFDWm(-2)). Although macroalgae and seagrasses are recognised as an alternative refuge against predation for amphipods, the high abundance of amphipods in SAV suggests that macroalgae represent a habitat that provides greater food availability. Pink shrimp Farfantepenaeus duorarum (Burkenroad, 1939) consumption rate (Mo) of epibenthic amphipods was experimentally evaluated. Mo intensifies as prey density increases and varied from 0.39 to 2.39 mg AFDWh(-1). Predation efficiency of F. duorarum on epibenthic amphipods was also evaluated in four artificial habitats with different physical complexity: soft-bottom substrates (SBS), small woody debris (SWD), seagrasses with densities of 300 and 1200 shootsm(-2) (S300 and S1200, respectively), macroalgae (MA), and at two prey densities (962 and 2406 indm( 2)). Amphipod consumption rate by F. duorarum varied from 1.20 to 2.07 indh(-1) in S1200 and MA, respectively. Habitat complexity had a significant effect on consumption rate, but prey density did not. Habitat physical complexity and predation efficiency maintained an inverse and a non-linear relationship. Presumably, the decrease in predation efficiency in association with the habitat complexity is due to the differential refuge value of these habitats. However, predation efficiency may also be influenced by either the microhabitat use by amphipods, the shrimp's dependence on seagrasses, or by differences in habitat value caused by the diel behavioural distribution pattern of amphipods and shrimp. Both field and experimental results highlight the importance of evaluating the relative value of tropical estuarine habitats through the study of the relationship between habitat physical complexity and predator-prey interactions. They also emphasise that inherent biological and ethological factors of the predator and prey involved, coupled to spatial and temporal variations in the habitat, should also be considered. PMID- 11018236 TI - Genetic structure of populations of the red sea urchin, Strongylocentrotus franciscanus. AB - Population subdivision was evaluated in the red sea urchin, Strongylocentrotus franciscanus, using DNA sequence data from 134 adult individuals collected in 1995 and 1996. On average 22 individuals were sequenced from six geographic locations between Alaska and Baja California (N=134), nearly the full extent of the species range. DNA sequence data was obtained from direct sequencing of a 273 base pair region of the bindin gene, which encodes a sperm fertilization protein. Results indicate that bindin is sufficiently polymorphic to serve as a genetic marker. We identified 14 unique alleles present in the entire range sampled with a maximum of eight alleles at a specific site. Mean pairwise comparison of the 14 unique alleles indicates moderate sequence diversity (p-distance=1.06). Although there is conflicting evidence to suggest that Alaska populations may deviate from the Hardy-Weinberg expectations, analysis of bindin genotype frequencies indicate that it is not possible to reject the null hypothesis of random mating throughout the species range. The results of a chi-square test with pooling conform to Hardy Weinberg expectations for all populations (P>0.05) except for the Alaska population (P=0.037). Inbreeding coefficients are consistent with this result and suggest that for the bindin locus, there is high gene flow. These results are compared with previously published results of genetic substructuring in sea urchins to examine relationships among population structure, dispersal potential and biogeography. PMID- 11018237 TI - Molecular and morphologic approaches to discrimination of variability patterns in chub mackerel, Scomber japonicus. AB - The systematic status and the evolutionary biology of chub mackerel (Scomber japonicus) in the South West Atlantic Ocean is confusing with an unknown degree of genetic differentiation and reproductive isolation between units. Simultaneous genetic and morphologic analyses were made on 227 fish collected from two areas of the South West Atlantic Ocean and one from the Mediterranean Sea. The genetic analysis was based on 36 protein-coding loci, 16 of which were variable. The morphologic analyses include six morphometric length measurements and a meristic character. Correspondence between genetic and morphologic variability patterns indicates isolated Mediterranean and Southwest Atlantic subgroups of S. japonicus and, less clearly, possible additional divergence in two regional stocks within the latter group. The most conservative approach to management is to manage the stocks independently of one another. PMID- 11018238 TI - Persistence and transport of fauna on drifting kelp (Macrocystis pyrifera (L.) C. Agardh) rafts in the Southern California Bight. AB - Drifting rafts of Macrocystis pyrifera may connect isolated kelp forests in the Southern California Bight. To determine which species might utilize this dispersal mechanism, faunal samples from natural drifting rafts and attached M. pyrifera plants were collected during five cruises between March 1995 and December 1997. These rafts, which can be considered as floating islands, were aged and the macroinvertebrate assemblage enumerated. There was no significant relationship between raft age and species richness, or between species richness and distance offshore, which contrasts with predictions based on island biogeography. Species richness, however, was related to raft weight. Patterns of species presence and density were investigated relative to raft age for the species most frequently associated with rafts. Only one species, the isopod Idotea resecata, was found on all sampled rafts. Some species increased in frequency with raft age and others decreased, but only one relationship, a decline in the frequency of the anemone Epiactis prolifera with raft age was significant. When species density was examined over all cruises, only I. resecata had a significant change in density (an increase) with raft age, but additional significant relationships were found when species density patterns were considered by cruise. The results of all the tests were combined to provide a measure of "raft success". Nine of the most frequent 19 species had a positive score, indicating a favorable response to rafting, seven were unaffected, and two species had negative responses to rafting. Extinction times were calculated using species abundance and raft age relationships. Two species (E. prolifera and Paracerceis cordata), were predicted to persist on rafts for only about 100 days, which is the maximum estimated raft lifetime. All other species were predicted to persist for longer periods if the rafts floated longer. Kelp fauna that begin rafting appear to be largely unaffected by rafting, and hence dispersal on kelp rafts is possible for many members of the kelp forest community. PMID- 11018239 TI - Age of drifting Macrocystis pyrifera (L.) C. Agardh rafts in the Southern California Bight. AB - Macrocystis pyrifera plants that detach from the substratum float to the surface and, if they do not become entangled or wash immediately to the shore, may drift at the surface for an unknown period of time. These rafts provide habitat for a variety of coastal and pelagic fauna. The distances dispersed and the period available for species to utilize these habitats, however, depend on the longevity of the raft and methods for determining the age of rafts are unknown. A method to age drifting M. pyrifera rafts based on a change in length of blades (BL) following detachment is validated here. This technique determines the period of time since detachment and not the actual age of the plant. In general, average BL decreases from initial attached values of 50-60 to about 0 cm, when rafts sink. The rate of aging, or deterioration of BL, is related to water temperature, and sets the period a raft stays afloat. Maximal estimates of ages of rafts were between 63 and 109 days, depending on the exact method used. Their lifetime will limit the distances dispersed by kelp rafts in Southern California, and this methodology will be useful for determining the temporal patterns of abundance of fauna associated with rafts. PMID- 11018240 TI - Effects of sediment type and intra- and interspecific competition on growth rate of the marine snails Hydrobia ulvae and Hydrobia ventrosa. AB - The combined effects of sediment grain size and intra- and interspecific competition on growth rate of Hydrobia ulvae and H. ventrosa were investigated in a 2-month laboratory experiment. Individuals of H. ulvae grew faster than H. ventrosa at all competition intensities. Growth rate did not differ between fine- and coarse-grained sediments in any of the species. This suggests that other factors than the sediment quality cause the often observed size differences between Hydrobia snails from these sediment types in nature. Competition from 1.6 to 1.9 times larger competitors affected growth of the smaller target snails. Competition between the species was asymmetrical and, in both species, per capita effect of competition was stronger from H. ulvae than from H. ventrosa, regardless of sediment type. From growth, Lotka-Volterra competition coefficients were estimated to 0.2 from H. ventrosa on H. ulvae and 1.8 from H. ulvae on H. ventrosa. Since competition between the species seems to be independent of sediment grain size, the shift in numerical dominance in nature, from H. ulvae on coarse-grained sediment to H. ventrosa on fine-grained, could not be explained with different competitive ability in different environments. PMID- 11018241 TI - Eclampsia and postpartum cerebral angiopathy. PMID- 11018242 TI - Relationship between disease activity and dose-response relationships with beta interferon therapies in the treatment of multiple sclerosis. AB - Beta-interferon (IFNbeta) therapy in patients with multiple sclerosis has proven efficacious in the treatment of multiple sclerosis (MS). Numerous studies have demonstrated a decrease in relapse rates, decreases in new gadolinium enhanced lesions, slowed accumulation of disability, and decreases in the accumulation of lesion burden on T2 weighted MRI scans. There has been a heterogeneity of responsiveness to IFNbetas in individual patients. Responsiveness to IFNbeta therapy is discussed in view of pre-existing disease activity. Disease activity can be objectively measured using the frequency and volume of new gadolinium enhancing lesions. Disability may be dependent upon lesion location, frequency, and by the intensity of inflammation within lesions. The presence of a dose response relationship with IFNbeta therapy in MS is supported by available data. Evidence is reviewed supporting the argument that responsiveness to IFNbeta therapy depends, in part, on pre-existing disease activity. PMID- 11018243 TI - In vivo evaluation of remyelination in rat brain by magnetization transfer imaging. AB - The aim of this work was to assess quantitatively and qualitatively the ability of magnetization transfer imaging to follow in vivo remyelination. Demyelination lesions were induced in rats by the injection of L-alpha-lysophosphatidylcholine stearoyl into the corpus callosum and imaging was performed in vivo on a 4.7 Tesla system at different time points. The percentage of magnetization transfer ratio (MTR) decrease was calculated for each animal. To evaluate the MTR findings for remyelination, myelin was quantitated by histological analysis of the lesion size and counting the number of remyelinating axons. An MTR decrease was observed when demyelination was present at 7 days after injection. During the remyelinating phase between day 30 and 40 after injection, contralateral values almost complete returned to normal, thus indicating remyelination. Histologically, at days 30 and 40 after injection, the lesion area was reduced in size and the axons were surrounded by a thin myelin sheath, indicating the remyelination process. Statistical analysis showed that the profile of MTR values was significantly correlated with the course of remyelination. All the MTR changes show a correlation with both myelin damage and repair. In conclusion, the study of the MTR profile in this myelin lesion model demonstrates in vivo the loss of myelin and the presence of spontaneous remyelination. This methodological approach which can also be applied to multiple sclerosis patients to show demyelination, should prove helpful to determine the degree of spontaneous and therapeutically induced remyelination in multiple sclerosis lesions, and thus to validate therapeutic treatments for myelin repair. PMID- 11018244 TI - Comparison of the acute-phase response in patients with ischemic stroke treated with high-dose heparin or aspirin. AB - Experimental studies have suggested that unfractionated heparin (UH) has antiinflammatory properties. It is unknown whether UH also has these properties in patients with acute ischemic stroke. Within 12-24 h of treatment onset we measured the acute-phase response as reflected by the erythrocyte sedimentation rate (ESR) and total number of leukocytes (x10(9)/l) in 706 consecutive patients with acute ischemic stroke treated with full-dose UH (n=450), or 300 mg/day aspirin (n=256). Clinical outcome (Mathew scale) at hospital discharge and the effect of factors such as treatment (UH and aspirin), and acute phase response were assessed using multivariate analyses adjusted for baseline confounders and incident complications. Separate models were created for patients with lacunar and nonlacunar stroke. Whereas there were not differences at baseline between the two treatment groups, total leukocyte counts (8. 0+/-4.1 vs. 8.6+/-3.2, P<0.01) and ESR (21.7+/-20.9 vs. 25.2+/-22.9, P<0.05) were statistically significantly lower in patients treated with UH. This effect of UH was more accentuated in patients with nonlacunar stroke. Overall, leukocytes (7.2+/-2.3 vs. 8.4+/-4.0, P<0. 01), and ESR (15.7+/-17.2 vs. 24.3+/-22.2, P=0.0001) were lower in patients with complete early recovery and this effect was restricted to patients with nonlacunar stroke. Whereas baseline impairment, symptomatic bleeding and stroke recurrence were independent negative outcome predictors, the use of UH was positively associated with early recovery in all patients. This study shows that full-dose UH reduces the acute-phase reaction that follows ischemic stroke more effectively than aspirin. The prognostic implications of such effect seem more notable in patients with nonlacunar stroke. PMID- 11018245 TI - The effect of the nitric oxide donor glyceryl trinitrate on global and regional cerebral blood flow in man. AB - Despite their potential use as cerebral vasodilatory agents there are few studies of the effect of nitric oxide (NO) donors on the cerebral circulation in non anaesthetised man. We determined the effect of the NO donor glyceryl trinitrate (GTN) at clinically relevant doses on global and regional cerebral blood flow (CBF) in healthy non-anaesthetised volunteers, using H(2)(15)O PET, ultrasonic colour velocity flow imaging of carotid artery flow, and transcranial Doppler (TCD) of middle cerebral artery velocities (MCAv). Three rates of GTN infusion (0.1, 0.4, 1.0 microg/kg/min) were used. There was no significant change in common or internal carotid artery flow following GTN administration although a dose dependent fall in MCAv post GTN was observed. There was no significant change in either global or regional CBF following GTN. Thus intravenous GTN at therapeutic doses in awake humans does not alter global or regional CBF. However it does produce basal cerebral artery vasodilatation as evidenced by a fall in MCAv in the absence of a change in internal carotid artery flow. PMID- 11018246 TI - Kearns-sayre syndrome: oncocytic transformation of choroid plexus epithelium. AB - Kearns-Sayre syndrome (KSS) is a sporadic multisystem disorder due to a defect of oxidative phosphorylation and associated with clonally-expanded rearrangements of mitochondrial DNA (mtDNA) deletions (Delta-mtDNAs) and/or duplications (dup mtDNAs). To gain further insight into the pathogenesis of CNS dysfunction in KSS, we studied the choroid plexus from two autoptic cases using in situ hybridization (ISH) of mtDNA, and immunohistochemistry to detect mtDNA and nuclear DNA-encoded subunits of the respiratory chain. Neuropathological examination of both cases showed oncocytic transformation of choroid plexus epithelial cells. In the same cells, ISH demonstrated that the predominant species of mtDNA were Delta-mtDNAs, and immunohistochemistry showed a decreased expression of mtDNA-encoded proteins. We suggest that mitochondrial abnormalities due to the presence of abundant Delta mtDNAs in the choroid plexus play an important role in causing the increased cerebrospinal fluid (CSF) protein and reduced folic-acid levels that are characteristic of KSS. PMID- 11018247 TI - Does high-dose interferon beta-1b improve clinical response in more severely disabled multiple sclerosis patients? AB - Prospective clinical open label follow-up of 52 multiple sclerosis patients treated with interferon beta-1b. After 18 months of treatment at standard 8 million international units (MIU) dose, subcutaneously on alternate days, IFNB dose was increased to 12 MIU in ten clinically non-responder patients. Eighteen months after, mean exacerbation rate, number and severity of exacerbations and number of patients with exacerbations or requiring corticosteroid treatment significantly improved, becoming similar to those of IFNB responders, always treated with 8 MIU. Baseline EDSS score of non-responders was higher than that of responders. Frequency and severity of adverse events were trending higher and dropout frequency higher in 12 MIU IFNB-treated patients. PMID- 11018248 TI - Alexia caused by a fusiform or posterior inferior temporal lesion. AB - We evaluated the alexia and agraphia of three patients with different lesions using Japanese kanji (morphograms) and kana (phonograms) and made a lesion-to symptom analysis. Patient 1 (pure alexia for both kanji and kana and minor agraphia for kanji after a fusiform lesion) made more paragraphic errors for kanji, whereas patient 2 (alexia with agraphia for kanji after a posterior inferior temporal lesion) showed severe reading and writing disturbances and more agraphic errors for kanji. Brodmann Area 37 was affected in both patients, but in patient 2 the lesion was located lateral to that in patient 1. Patient 3 showed agraphia without alexia after restricted lesion to the angular gyrus. We believe that pure alexia (patient 1) results from a disconnection between the medial fusiform gyrus and posterior inferior temporal area (the lateral fusiform and inferior temporal gyri), whereas alexia with agraphia for kanji (patient 2), corresponding to lexical agraphia in Western countries, results from damage to the posterior inferior temporal area, in which whole-word images of words are thought to be stored. Furthermore, restricted lesion in the angular gyrus (patient 3) does not produce alexia; the alexic symptom of "angular" alexia with agraphia may be the result of damage to the adjacent lateral occipital gyri. PMID- 11018249 TI - Correlation between coefficient of variation of choice reaction time and components of event-related potentials (P300): effect of benzodiazepine. AB - We studied the relationship between accuracy in the cognitive process and components of event-related potentials (P300) in 21 young and healthy subjects. Benzodiazepine was used to manipulate the cognitive state of the subjects. We recorded the serial changes in P300, choice reaction time (CRT), and error ratio (ER) before and after oral administration of 0.4 mg of alprazolam. After administration, the coefficient of variation of CRT tended to decrease in nine subjects (group I) and increase in 12 subjects (group II). Prolongation of the P300 latency was observed in all subjects after treatment; however, such change was more predominant in group II than in group I. In group I, there was no error and no significant difference in P300 amplitude before and after administration. In group II, alprazolam significantly reduced P300 amplitude and increased ER. Our results suggest that the accuracy and P300 amplitude were preserved when the central nerve system managed to reduce fluctuations in CRT but P300 amplitude diminished and the error ratio increased following deterioration of these processes. PMID- 11018250 TI - Blood oxidative stress in amyotrophic lateral sclerosis. AB - It has been suggested that amyotrophic lateral sclerosis (ALS), a neurodegenerative disorder resulting in motor neuron death, is associated with oxidative damage induced by free radicals. Our study aimed to get an assessment of the blood oxidative stress status in a population of 167 ALS patients (aged 59+/-13 years), treated or not with riluzole, compared with 62 age-matched healthy control subjects (aged 60+/-11 years) simultaneously included in the study. We determined the level of plasma lipid peroxidation (thiobarbituric acid reactive substances, TBARS); the status of the major lipophilic plasma antioxidant defenses (vitamin E, vitamin A and beta-carotene); the activities of erythrocyte Cu,Zn-superoxide dismutase (Cu,Zn-SOD) and of plasma and erythrocyte glutathione peroxidase (GSH-Px). Plasma selenium was also determined as a trace element essential to the activity of the GSH-Px. In comparison with controls, we observed in ALS patients (mean+/-S.D.) significantly higher TBARS values (ALS=1.34+/-0.28 micromol/l; controls=1.11+/-0. 20 micromol/l) and a significant enhancement of the erythrocyte SOD activity (ALS=710+/-114 U/g Hb; controls=667+/ 93 U/g Hb). No differences were observed for selenium level, GSH-Px activity, plasma vitamin E, beta-carotene and vitamin A concentrations. These data confirm the presence of an oxidative stress in blood of ALS patients. The elevated plasma TBARS, without any deficiency in plasma lipophilic antioxidants such as vitamin E, vitamin A and beta-carotene, suggest an enhancement in the production of free radicals. No correlation was found in our study between the level of any of the blood oxidative stress markers and the disease duration. Comparison between patients treated or not with riluzole did not display any modification of the plasma TBARS concentration, but we observed a slight decrease of erythrocyte SOD activity in treated patients (treated=705+/-113 U/g Hb; not treated=725+/-118 U/g Hb), suggesting a possible activity of riluzole on the oxygenated free radical production. PMID- 11018252 TI - Neurological accidents caused by repetitive breath-hold dives: two case reports. AB - We report two Japanese male professional breath-hold divers (33 and 39 years of age) who experienced neurological disorders during repetitive dives to over 20 m of seawater. One patient had right homonymous hemianopsia, and the other presented with right hemiparesis with facial involvement and sensory deficit. In addition, they each had a history of neurological problems following such dives. Magnetic resonance images of their brains disclosed multiple T2-weighted hyperintensities corresponding to their neurological symptoms. Their brain lesions suggest a multiple cerebral infarction caused by occlusion of the cerebral arteries. We conclude that the repetitive deep breath-hold dives induced the brain involvement. PMID- 11018251 TI - Successful immunosuppressant therapy of severe progressive cerebellar degeneration and sensory neuropathy: a case report. AB - A 56 year old woman had a 19 month history of a severe subacute progressive cerebellar degeneration, peripheral sensory neuropathy, and urinary incontinence. She was confined to a wheelchair, needed assistance with eating, and her speech was almost unintelligible. No underlying cancer was found despite repeated investigations, and no autoantibodies were demonstrated. She received a 3-month course of intensive immunosuppressant therapy with intravenous immunoglobulin 400 mg/kg per day for 5 days every month, oral cyclophosphamide 50 mg twice or three times a day to maintain the total lymphocyte count between 500 and 750/mm(3), and prednisone 60 mg per day. She experienced dramatic subjective and objective improvement. The dysarthria and the upper extremity dysmetria disappeared, and she regained the ability to write and cook. The lower extremity ataxia improved and she became able to walk with a cane. Urinary incontinence disappeared. A trial of intensive immunosuppressant treatment is worth considering in a patient with a clinical syndrome resembling paraneoplastic disorders, even if an underlying neoplasm and autoantibodies are not demonstrated. PMID- 11018253 TI - Parkinson's disease associated with argyrophilic grains clinically resembling progressive supranuclear palsy: an autopsy case. AB - A 70-year-old male began to show akinesia, rigidity of extremities, finger tremor, disturbed vertical external ocular movement, and nuchal dystonia, which progressed slowly. Brain CT scan and magnetic resonance images showed slight atrophy of the frontal lobe and slight enlargement of the lateral ventricles. Hasegawa's dementia rating scale-revised version gave a moderate score of 11/30 points. He died of pneumonia at the age of 76. The clinical diagnosis was progressive supranuclear palsy (PSP). However, there were no neuropathological characteristics of PSP. Neuropathologically, Parkinson's disease was diagnosed. In addition, many argyrophilic grains (ArGs) in the gray matter were stained, especially in the insula, amygdala, hippocampus, parahippocampal gyrus, lateral occipitotemporal gyrus, and substantia nigra, by the Gallyas-Braak method. We consider that ArGs could modify the symptoms of Parkinson's disease and that Parkinson's disease with ArGs may show a PSP-like clinical course. PMID- 11018254 TI - The human transcriptional repressor protein NAB1: expression and biological activity. AB - The zinc finger protein early growth response 1 (Egr-1) is a transcriptional activator involved in the regulation of growth and differentiation. Egr-1 has a large activating domain and three zinc finger motifs that function as a DNA binding region. We show here that a third functional domain of the Egr-1 protein, localized between the extended activation domain and the zinc finger DNA binding region, acts as a transcriptional repressor domain when fused to a heterologous DNA binding domain (DBD). Through protein-protein interaction this inhibitory domain of Egr-1 brings the transcriptional corepressor NAB1 in close proximity to the transcription unit. NAB1 is expressed ubiquitously in human cell lines as shown by RNase protection mapping. Overexpression studies revealed that NAB1 is able to completely block transcription mediated by Egr-1. In addition, the transcriptional repression activity of a fusion protein containing the inhibitory domain of Egr-1 and the DBD of the yeast transcription factor GAL4 was increased by overexpression of NAB1. A fusion protein consisting of the DBD of GAL4 and the coding region of human NAB1 repressed transcription from model promoters with engineered upstream GAL4 binding sites. The GAL4-NAB1 fusion protein functioned from proximal and distal positions indicating that NAB1 displays transcriptional repressor activity at any position within the transcription unit. Thus, the biological function of the inhibitory domain of Egr-1 is solely to provide a docking site for NAB1 via protein-protein interaction. PMID- 11018255 TI - Sequence analysis of the human glycoprotein hormone alpha-subunit gene 5' flanking DNA and identification of a potential regulatory element as an alu repetitive sequence. AB - The nucleotide sequence of the human glycoprotein hormone alpha-subunit (GPHalpha) gene 5'-flanking DNA was determined from -1637 to +49 relative to the cap site (+1). Comparison of the upstream sequence of the human gene with those of rhesus and mouse demonstrates regions with variable identity. When the 1.7 kb fragment was used to drive the expression of chloramphenicol acetyltransferase (CAT) in transiently transfected HeLa cells, it was found that CAT activity was elevated about 3-fold when the fragment was truncated from -1637 to -846, suggesting the presence of a negative regulatory element in the distal 5' flanking DNA. This overlaps an Alu repetitive sequence (ARS) located between nucleotides -1330 and -1007. Gel mobility shift and DNase protection analyses identified a protein binding site centered around -1100 in the ARS second monomer. The GPHalpha upstream ARS was cloned in both orientations in positions upstream and downstream from the bacterial CAT gene under control of the herpes simplex virus thymidine kinase (tk) promoter. DNA-mediated transient transfection of these plasmids revealed a marked inhibition (79-82%) of CAT production by the ARS when it was cloned upstream from the tk promoter and in the same orientation as that found in the GPHalpha 5'-flanking DNA. Smaller decreases (29-57%) were produced by the ARS cloned upstream from the tk promoter in the reverse orientation. In marked contrast, the Alu repetitive element had little or no effect when cloned in either orientation downstream from the tk-CAT gene. Introduction of a second ARS downstream from the CAT reporter gene in vectors already containing an ARS upstream from the tk promoter significantly reduced the strong negative effect elicited by the upstream repetitive element. When compared to the Blur 8 Alu element, the GPHalpha upstream ARS differs markedly with respect to its effect on tk-CAT expression in transient assays and as a substrate for DNA binding proteins present in HeLa nuclear extracts. Together, the transient expression results demonstrate that ARS elements can influence expression of nearby class II promoters. The extent of this effect depends on element position and orientation, cell type, the particular ARS (e.g., GPHalpha or Blur 8), and whether copies were present both upstream and downstream from the transcription unit. PMID- 11018256 TI - Organization and expression of the human zo-2 gene (tjp-2) in normal and neoplastic tissues. AB - One of the tight junction components, zonula occludens protein 2 (ZO-2), is expressed as two isoforms, ZO-2A and ZO-2C, in normal epithelia. In pancreatic adenocarcinoma of the ductal type ZO-2A is absent, but none of the common mechanisms of gene inactivation is responsible for lack of ZO-2A expression. In the current study, we report the complete organization of the human zo-2 gene (tjp-2), its alternative splicing, and its expression in normal and neoplastic tissues of several organ sites. In addition to pancreatic adenocarcinoma, ZO-2 was found to be de-regulated in breast adenocarcinoma, but not in colon or prostate adenocarcinoma. The latter are considered to be of acinar rather than ductal type. Thus, our data indicate the importance of zo-2 (tjp-2) gene regulation in ductal cancer development and should help to understand the defects of intercellular interactions, critical for suppressing the malignant phenotype. PMID- 11018257 TI - The DNA-binding domain of the gene regulatory protein AreA extends beyond the minimal zinc-finger region conserved between GATA proteins. AB - The AreA protein of Aspergillus nidulans regulates the activity of over 100 genes involved in the utilisation of nitrogen, and has a limited region of homology with the vertebrate family of GATA proteins around a zinc finger (Zf) motif. A 66 amino acid (a.a.) residue fragment (Zf(66)) corresponding to the zinc finger, a 91 a.a fragment (Zf(91)) containing an additional 25 a.a. at the C-terminus, and a much larger 728 a.a. sequence (3'EX) corresponding to the 3'exon have been over expressed as fusion proteins in E. coli and purified. The DNA-protein complexes formed by these proteins have been examined by gel retardation analysis. The 91 a.a. protein forms a discrete shifted species with a GATA-containing DNA fragment with high affinity (K(d)=0.15 nM), whereas the 66 a.a. protein has very low ( approximately microM) affinity for the same sequence. The results show that the region of AreA required for high affinity DNA binding extends beyond the zinc finger motif that is homologous to GATA-1, requiring in addition a region within the 25 a.a. sequence C-terminal to the zinc finger. Using hydroxyl radical and ethylation interference footprinting, the minimal Zinc finger protein (Zf(66)) shows no appreciable interference effects whereas Zf(91) shows much stronger interference effects, identical to those of the larger protein. These effects extend over sequences up to two nucleotides either side of the GATA site, and indicate contacts additional to those observed in the three-dimensional structure of the complex of the minimal zinc-finger protein with DNA. We suggest that these additional contacts are responsible for the enhanced DNA binding affinity of the extended zinc-finger protein Zf(91). PMID- 11018258 TI - Molecular cloning of the murine PHEX gene promoter. AB - We report the novel cloning of the murine PHEX promoter, the gene that is mutated in X-linked hypophosphatemic rickets (XLH). Four promoter/reporter gene constructs, -133/+104, -542/+104, -1061/+104, and -2866/+104, showed significant luciferase activity (4.9-13.2-fold over background) when transfected into rat osteogenic sarcoma (UMR-106) cells. PMID- 11018259 TI - Molecular cloning and expression patterns of three putative functional aldehyde oxidase genes and isolation of two aldehyde oxidase pseudogenes in tomato. AB - The final steps in the biosynthesis of the plant hormones abscisic acid (ABA) and indole-3-acetic acid (IAA) have been shown to be catalyzed by aldehyde oxidases (AO). We have cloned three putative functional AO genes (TAO1, TAO2 and TAO3) and two putative AO pseudogenes (TAO4 and TAO5) in tomato. The TAO1 cDNA described here includes the correct amino terminus of the encoded TAO1 protein and is different at the 5'-end from the TAO1 sequence in GenBank (accession number U82558). Northern analysis shows that TAO1 is expressed mainly in vegetative tissues and TAO2 is expressed in both vegetative and reproductive tissues. TAO3 expression was not detectable by Northern hybridization. These results suggest that each AO may play different roles in the regulation of tomato growth and development. PMID- 11018260 TI - Chromosomal organization and localization of the human histone deacetylase 5 gene (HDAC5). AB - Histone deacetylases (HDACs) are important participants in the remodeling of chromatin structure and in the regulation of eukaryotic proliferation and differentiation. We have isolated and characterized the human HDAC5 genomic sequence, which spans a region of 39,138 bp and which has one single chromosomal locus. Determination of the exon-intron splice junctions established that HDAC5 is encoded by 26 exons ranging in size from 22 bp (exon 1) to 285 bp (exon 12). Characterization of the 5' flanking genomic region revealed that the human HDAC5 promoter lacks both the canonical TATA and CCAAT boxes. The human HDAC5 mRNA encodes a 1122 aa protein with a predictive molecular mass of 121.9 kDa and an isoelectric point of 5.84. Fluorescence in situ hybridization analysis localized the human HDAC5 gene to chromosome 17q21, a region which is characterized by frequent gains and losses of chromosomal material in several types of cancer. PMID- 11018261 TI - Isolation of a cDNA for a novel human RING finger protein gene, RNF18, by the virtual transcribed sequence (VTS) approach(1). AB - We have recently developed a novel database system, designated as the virtual transcribed sequence (VTS) which efficiently extracts many genes from public human genome databases, and tested the feasibility of this novel computational approach (N. Miyajima, C. Burge, T. Saito, Biochem. Biophys. Res. Commun. 272 (2000) 801; http://host45.maze.co.jp/vts/). In this study, using the VTS approach, we isolated a cDNA for a novel human gene with RING finger motif (C(3)HC(4)), which is not deposited in public EST databases. The isolated cDNA clone is 2163 bp in length, and contains an open reading frame of 452 amino acids. We designated the novel gene as RNF18. A database search showed that the RNF18 gene had the moderate similarity to SS-A/Ro52 protein, which is a ribonucleoprotein reactive with autoantibodies in patients with Sjogren's syndrome and systemic lupus erythematosus. Tissue distribution analyses by Northern blot and RT-PCR methods demonstrated that the RNF18 messenger RNA was preferentially expressed in testis. The exon-intron boundaries of RNF18 gene were determined by aligning the cDNA sequence with the corresponding genome sequence. The isolated cDNA consists of eight exons that span about 11 kb of the genome DNA. The precise chromosomal location of the RNF18 gene was determined by PCR based radiation hybrid mapping, and the gene was located to centromere region of chromosome 11 between markers NIB1900 and D11S1350. Taken together, the VTS approach should provide a novel cDNA cloning strategy for isolating unidentified genes, which are not found even in EST databases but are detectable computationally. PMID- 11018262 TI - Pregnancy-associated plasma protein-E (PAPP-E). AB - A full-length cDNA encoding a novel human protein was cloned from placenta cDNA. The corresponding 1542 amino acid protein sequence was termed 'pregnancy associated plasma protein-E' (PAPP-E) as it shows a 62% homology to the human pregnancy-associated plasma protein-A (PAPP-A) that is a diagnostic marker for trisomies, especially Down syndrome. The conserved domain structure contains five motifs related to the short consensus repeats of complement proteins and selectins, three motifs related to the lin-notch motifs of proteins regulating early tissue differentiation, and a putative zinc-binding motif and active site of the metzincin-superfamily of metalloproteases. The PAPP-E gene was localized to chromosome 1q23-25. Northern blot analysis showed that PAPP-E is predominantly expressed in placenta. PMID- 11018263 TI - Characterisation of the human plunc gene, a gene product with an upper airways and nasopharyngeal restricted expression pattern. AB - Here we report the cloning and characterization of the human homologue of plunc, a murine gene expressed specifically in the upper airways and nasopharyngeal regions. The human plunc cDNA codes for a leucine-rich protein of 256 amino acids which is 72% identical to the murine protein. RNA blot analysis suggests that expression of plunc is restricted to the trachea, upper airway, nasopharyngeal epithelium and salivary gland. The human plunc gene contains nine exons and is localised to chromosome 20q11.2. The unique expression pattern of the human plunc suggest that it may prove a useful model gene with which to study the regulatory mechanisms which direct expression of genes specifically to the upper airways. PMID- 11018264 TI - Gene regulation of Wiskott-Aldrich syndrome protein and the human homolog of the Drosophila Su(var)3-9: WASP and SUV39H1, two adjacent genes at Xp11.23. AB - The region Xp11.23 is a gene-rich, light giemsa-staining segment on the short arm of the X chromosome. In this study, we have characterized the transcriptional regulatory elements in this interval for two adjacent genes: SUV39H1, a regulator of chromatin organization, and the Wiskott-Aldrich syndrome protein (WASP). The WASP gene exhibits two alternate promoters, both of which demonstrate transcription factor binding elements specific to blood cell lineages. Reporter gene expression analyses indicate that both WASP promoters show high levels of expression in different hematopoietic cell lines. The human homolog of the Drosophila Su(var)3-9 gene was identified by sequence analysis of the region downstream from WASP. SUV39H1 is ubiquitously expressed, and the promoter sequence consists mostly of general transcription factors. The presence of putative binding sites for GAGA and Adf1 transcription factors may indicate a cross regulatory mechanism with other chromatin regulators. PMID- 11018265 TI - Identification of a novel FAS/ER-alpha fusion transcript expressed in human cancer cells. AB - A semi-nested reverse transcriptase-polymerase chain reaction (RT-PCR) was applied to evaluate the presence of estrogen receptor-alpha (ER-alpha) in human prostate cancer cells. Unexpectedly, a novel fatty acid synthase (FAS)/ER-alpha fusion transcript was identified, in which the N-terminus of FAS was fused in frame with the C-terminus of ER-alpha. The existence of the FAS/ER-alpha transcript was further confirmed by RT-PCR analysis using various sets of amplification primers and different reverse-transcribed primers in the presence of dimethyl sulfoxide to eliminate the secondary structure of RNA. The predicted FAS/ER-alpha protein would contain largely domain I of FAS and the entire ligand binding domain of ER-alpha. The FAS/ER-alpha was expressed in a variety of human cancer cell lines including prostate, breast, cervical and bladder cancer cell lines. Our data suggest that the presence of FAS/ER-alpha may complicate the FAS and the ER-alpha signalling pathway. PMID- 11018266 TI - Rat homologue of the human M(r) 110000 antigen is the protein that expresses widely in various tissues. AB - The rat homologue of the human M(r) 110000 antigen, which cross-reacts with anti carcinoembryonic antigen antibodies, was isolated from a rat lung cDNA library. The deduced amino acid sequence revealed a signal peptide, cysteine-rich and immunoglobulin-like region, serine-threonine region, and N-glycosylation sites in the extracellular portion. Northern blot analysis demonstrated a wide distribution of the mRNA in adult rat tissues and A10 rat vascular smooth muscle cells. Therefore, the rat homologue of the human M(r) 110000 antigen may be a receptor or a cell adhesion molecule rather than a specific carcinogenic antigen. PMID- 11018267 TI - Optimising recovery of Campylobacter spp. from the lower porcine gastrointestinal tract. AB - To determine the incidence of campylobacters in Northern Ireland pigs, ileal contents and anal swabs were taken shortly after death. Direct streaking onto Preston agar, and modified charcoal cefoperazone desoxycholate agar (mCCDA), were compared, as was enrichment in selective broths prior to streaking onto the corresponding solid medium. For anal swabs direct plating on mCCDA was most efficient, with 100% of samples positive, whilst for ileal contents enrichment in mCCD broth was best with 86% of samples positive. Although only 34% of ileal samples enriched in Preston broths were positive they yielded three species not isolated from mCCD broth, and hence indicated that some pigs were infected by at least two species of Campylobacter. Overall, the number of samples found to contain campylobacters, and the range of species isolated, was seen to be markedly affected by both the choice of selective medium and the isolation procedures. PMID- 11018268 TI - Contrast-enhanced immunoelectron microscopy for Helicobacter pylori. AB - Since a method of contrast enhancement for immunoelectron microscopy has not been available in bacteriology, the morphological localization of proteins of Helicobacter pylori is not well known. In this report, we established a method of contrast enhancement in immunoelectron microscopy in this organism. Immunostained ultrathin sections are stained with a mixture of alcian blue and osmium tetroxide prior to staining with uranyl acetate. This method of staining provided good contrast enhancement of the bacterial cell wall and membrane without any loss of immunolabeled gold particles on the ultrathin section. PMID- 11018269 TI - A fluorescence microscopy based genetic screen to identify mutants altered for interactions with host cells. AB - The study of microbial intracellular pathogenesis has benefited from the application of immunofluorescence microscopy to characterize interactions of the pathogen with host cells. Unfortunately, immunofluorescence microscopy is impractical for screening the large number of bacterial mutants necessary to represent the entire genome of the pathogen. Screening has been limited due to the lack of materials suitable for high-throughput processing (e.g. 96-well plates) that also possess the optical features needed for high resolution fluorescence microscopy. Recently marketed 96-well Special Optics (SO) plates provide both the 96-well template ideal for high-throughput analysis and optical features suitable for fluorescence microscopy. Until this work, mutants needed for the study of a fluorescence-based virulence phenotype could not be obtained by direct screening approaches. In this study, SO plates were used to examine 11520 individual Salmonella typhimurium MudJ mutants for the loss of the ability to disrupt host cell endocytic compartments. The direct application of the fluorescence phenotype for screening allowed us to obtain a set of mutants to characterize the formation of lysosomal membrane glycoprotein (lgp) containing tubules upon Salmonella infection of HeLa epithelial cells. This approach will facilitate the characterization of a wide range of microbial phenotypes detectable by fluorescence microscopy. PMID- 11018270 TI - A simple and sensitive method to extract bacterial, yeast and fungal DNA from blood culture material. AB - This study investigated the various commercially available kits and 'in-house' methods to extract DNA from Gram-negative and Gram-positive bacteria, yeast and fungal agents in commonly employed blood culture material. The main methods investigated were as follows; Qiagen QIAmp Blood kit, Roche high PCR template preparation kit, Puregene DNA extraction kit, boiling, glass beads/sonication and wash/alkali/heat lysis. The results indicated that a simple wash/alkali/heat lysis method was the most sensitive, reproducible, simple and cost-effective extraction method. This was the only method which removed any PCR inhibitors and inherent DNA which existed in virgin BacT/Alert aerobic, anaerobic and paediatric blood culture material. Contaminating microbial DNA from Lactococcus lactis or Bacillus coagulans was identified in all batches of BacT/Alert FAN aerobic blood culture material examined. PMID- 11018271 TI - Rapid spectrophotometric determination of 2,4,6-trinitrotoluene in a Pseudomonas enzyme assay. AB - Although TNT (2,4,6-trinitrotoluene) and its degradation products can be quantified by HPLC, this method is not suitable for simultaneous analyses of the numerous samples typically encountered in enzyme studies. To solve this problem, we developed a simple and rapid spectrophotometric assay for TNT and tested the procedure using partially purified nitroreductase(s) from a Pseudomonas aeruginosa isolate, which transformed TNT in the culture medium. In highly alkaline solution, TNT (pK(a)=11.99) exhibits significant absorbance at 447 nm, while major metabolites, 2-amino-4, 6-dinitrotoluene (2ADNT), 4-amino-2,6 dinitrotoluene (4ADNT), and 2,6-diamino-4-nitrotoluene (2,6DANT) display no absorbance at this wavelength. Assay mixtures of TNT, Tris-HCl buffer, a reductant, and the enzyme(s) were analyzed by measuring absorbance 4 min after adjusting the pH to 12.2. TNT transformation to colorless metabolites was linear with respect to protein and substrate concentrations. Using the assay, we determined that TNT nitroreductase(s) from the isolate required an electron donor and preferred NADH to NADPH. TNT transformation increased when NAD was recycled to NADH using glucose-6-phosphate (GP) and glucose-6-phosphate dehydrogenase (GPDH). Enzymatic transformation of TNT was completely inhibited by Cu(2+) (5 mM) and was partially inhibited by other divalent metallic cations. Because the assay is sensitive to ammonium sulfate, dithiothreitol, ascorbic acid, and sodium phosphate, extracts should be assayed in the absence of these components. PMID- 11018272 TI - A sensitive standardised micro-gel well diffusion assay for the determination of antimicrobial activity. AB - We have developed a highly sensitive micro-gel well diffusion assay for the determination of antimicrobial activity. In essence, the normal radial diffusion type assay was adapted to perform it in a microtiter plate. We compared our micro gel well diffusion assay to a radial diffusion assay and a microtiter broth dilution method, using gramicidin S as model antibiotic, and Micrococcus luteus as the indicator organism. The micro-gel well diffusion assay was as sensitive as the microtiter broth dilution method, and approximately twice as sensitive as the radial diffusion method. Data analysis to calculate minimum inhibitory concentration, 50% microbial growth inhibition and maximum inhibitory concentration was refined by generating dose-response curves with the software package Prism 3.0 (Graphpad Software Inc.). The minimum inhibitory concentrations, determined by the three methods, were significantly different (P<0. 001), highlighting the limitations involved in comparing data obtained from different methods. PMID- 11018273 TI - Discrimination among iron sulfide species formed in microbial cultures. AB - A quantitative method for the study of iron sulfides precipitated in liquid cultures of bacteria is described. This method can be used to quantify and discriminate among amorphous iron sulfide (FeS(amorph)), iron monosulfide minerals such as mackinawite or greigite (FeS(min)), and iron disulfide minerals such as pyrite or marcasite (FeS(2min)) formed in liquid cultures. Degradation of iron sulfides is performed using a modified Cr(2+) reduction method with reflux distillation. The basic steps of the method are: first, separation of FeS(amorph); second, elimination of interfering species of S such as colloidal sulfur (S(c) degrees ), thiosulphate (S(2)O(3)(2-)) and polysulfides (S(x)(2-)); third, separation of FeS(min); and fourth, separation of FeS(2min). The final product is H(2)S which is determined after trapping. The efficiency of recovery is 96-99% for FeS(amorph), 76-88% for FeS(min), and >97% for FeS(2min). This method has a high reproducibility if the experimental conditions are rigorously applied and only glass conduits are used. A well ventilated fume hood must be used because of the toxicity and volatility of several reagents and products. The advantage relative to previously described methods are better resolution for iron sulfide species and use of the same bottles for both incubation of cultures and acid degradation. The method can also be used for Fe/S stoichiometry with sub sampling and Fe analysis. PMID- 11018274 TI - An evaluation of chelex-based DNA purification protocols for the typing of lactic acid bacteria. AB - An easy and rapid protocol to extract DNA to be used as template for polymerase chain reaction (PCR) fingerprinting experiments from cultivable lactic acid bacteria (LAB) is proposed. Different procedures for rapid extraction of DNA by chelex (iminodiacetid acid) ionic resin were compared. Factors affecting the quality and reproducibility of PCR fingerprinting profiles were also investigated. Two out of three chelex-based protocols allowed to obtain DNA samples which, after PCR amplification, provided electrophoretic patterns comparable with those obtained by classical lysozyme and phenol-chloroform DNA extraction. A good level of reproducibility and consistency of the InstaGene procedure was verified. The procedure is fast, practical, and the DNA is of quality similar to that obtained by phenol-chloroform extraction. Although applied to a little number of LAB strains, chelex-based protocols are potentially applicable to a vast array of organisms and/or biological materials. PMID- 11018275 TI - Identification of cell wall deficient forms of M. avium subsp. paratuberculosis in paraffin embedded tissues from animals with Johne's disease by in situ hybridization. AB - M. avium subsp. paratuberculosis (M. paratuberculosis) is the causative agent of Johne's disease (JD) in ruminants leading to enormous economical losses in dairy and meat industries worldwide. During the subclinical stage of the disease, the infected animals are difficult if not impossible to detect by the available diagnostic tests including the PCR based ones. Although only considered an animal pathogen, cell wall deficient (CWD) forms of M. paratuberculosis have been isolated from patients with sarcoidosis and Crohn's disease (idiopathic diseases) in humans. Hence, the CWD form of this organism has been suspected to play a role in the pathogenesis of these diseases by persisting in the affected tissues and triggering a localized immune response and pathology. Differentiating between the CWD and acid-fast forms of this organism may lead to the determination of whether the CWD form is the pathogenic form in the subclinical cases of JD in animals and/or the etiologic agent for the above human diseases. To localize such organisms in tissue sections, CWD forms of mycobacteria were prepared in vitro and injected into beef cubes which were then formalin fixed and paraffin embedded. An in situ hybridization (ISH) technique, combined with the IS900 M. paratuberculosis-specific probe labeled with digoxigenin, was developed for the detection of nucleic acids specifically from the CWD forms but not their acid fast forms in tissue sections. Specificity was confirmed by the negative finding with an irrelevant probe and with control tissue preparations containing CWD cells of related mycobacteria and unrelated organisms. This ISH procedure provides a way to distinguish between the acid-fast and CWD forms of M. paratuberculosis and to localize them in tissue sections. ISH may prove useful to evaluate the significance of CWD forms of M. paratuberculosis in the pathogenesis of JD, Crohn's disease and sarcoidosis. PMID- 11018276 TI - An improved vector system for insertional gene inactivation inspired by the tmRNA tagging system of S. pneumoniae. AB - Insertional mutagenesis is a technique often used to inactivate genes in Streptococcus pneumoniae. Using conventional vectors, a 5' segment of the targeted gene remains under the control of the gene's authentic promoter following gene disruption. Thus, the expression of a functional peptide and the misinterpretation of results in consequence cannot be excluded. To circumvent this problem, we have developed a plasmid for insertional mutagenesis based on the tmRNA-tagging system of S. pneumoniae which ensures that any protein expressed after gene disruption is degraded. Insertional mutagenesis using this vector results in the targeted gene being tagged with a tmRNA-derived sequence coding for a proteolysis tag. Here we show that the translation product of a gene tagged by this method is not detectable by Western blotting, suggesting that the protein was degraded. This modified vector allows total inactivation of genes with a reliability that cannot be achieved by conventional vectors for insertional mutagenesis. This approach can be applied to other bacterial species. PMID- 11018277 TI - The addition of low numbers of 3' thymine bases can be used to improve the hybridization signal of oligonucleotides for use within arrays on nylon supports. AB - Oligonucleotide arrays can be used for the analysis of microbial nucleic acid. The addition of high numbers of dTTP to the 3' ends of oligonucleotides using terminal transferase has been shown to facilitate membrane binding. This paper demonstrates low numbers of thymine bases added to the 3' end of oligonucleotides during synthesis can improve hybridisation signal intensity where the signal seen with the unmodified oligonucleotides is poor. Thus, the addition of variable numbers of thymine bases to different oligonucleotides allows the production of oligonucleotide arrays producing strong interpretable hybridisation signals. PMID- 11018278 TI - The cleavage activation and sites of glycosylation in the fusion protein of Hendra virus. AB - Hendra virus (HeV) is an unclassified member of the Paramyxoviridae family that causes systemic infections in humans, horses, cats, guinea pigs and flying foxes. The fusion protein (F(0)) of members of the Paramyxoviridae family that cause systemic infections in vivo contains a basic amino acid-rich region at which the protein is activated by cleavage into two subunits (F(1) and F(2)). HeV F(0) lacks such a domain. We have determined the cleavage site in HeV F(0) by sequencing the amino terminus of the F(1) subunit and in view of the potential effect of glycosylation on the cleavage process have ascertained the sites at which F(0) is glycosylated. The results indicate that unlike other members of the family that replicate in cultured cells and cause systemic infections in vivo, cleavage of HeV F(0) occurs at a single lysine (reside 109) in the sequence Asp Val-Lys- downward arrow-Leu. Although HeV genotypically resembles members of the Respirovirus and Rubulavirus genera in having potential N-linked glycosylation sites in both the F(1) and F(2) subunits, we show that phenotypically HeV may more closely resemble members of the Morbillivirus genus that contain N-linked glycans only in the F(2) subunit. PMID- 11018279 TI - Bovine viral diarrhea virus induced apoptosis correlates with increased intracellular viral RNA accumulation. AB - Non-cytopathic (NCP) and cytopathic (CP) parent-daughter pairs are often isolated from cattle with bovine viral diarrhea virus (BVDV) induced mucosal disease. Alignment of these pair genomes revealed that genetic changes in CP BVDV involve the NS2-3 coding region and correlate with expression of NS3. However, additional mutations are present elsewhere in the genomes of these natural pairs, precluding unambiguous mapping of this function to the NS2-3 region. To evaluate this phenomenon in identical genetic backgrounds, we have constructed an NCP isogenic pair of the NADL by deletion of the cIns from NS2 region. The levels of viral protein synthesis in infected cells revealed no marked difference between the CP and the isogenic NCP BVDV mutant. In contrast, RNA accumulation in cells infected with CP virus was up to 25 times higher than that in cells infected with NCP BVDV. No significant difference in growth kinetics and viral yields were observed between the CP BVDV and the isogenic NCP pair. Analyses of additional NCP/CP parent-daughter field BVDV isolates revealed a similar pattern of macromolecular synthesis, suggesting the generality of this phenomenon. These results implicate increased levels of RNA accumulation in CP BVDV infected cells, along with the production of NS3 as potential contributors to viral cytopathogenicity. PMID- 11018280 TI - Nuclear localization and intramolecular cleavage of N-terminally deleted NS5A protein of hepatitis C virus. AB - The full-size NS5A (NS5A-F) of hepatitis C virus is localized in the cytoplasm despite the presence of a functional nuclear localization signal (NLS) in its C terminal region (amino acids (aa) 354-362). In the present study, we demonstrated that a short stretch of sequence near the N-terminus of NS5A (aa 27-38) masked the functional NLS, preventing NS5A from being transported to the nucleus. This sequence, referred to as an NLS-masking sequence, was distinct from a nuclear export signal, as it did not actively target a protein to the cytoplasm. We also found that other sequences located at either an N- (aa 1-21) or a C-terminal region (aa 353-447) were responsible for targeting NS5A to the cytoplasm. Western blot analysis of the transfected cells revealed that NS5A mutants that had been N terminally deleted by 66 aa or more were cleaved at a certain cleavage site, generating a common fragment of ca. 40 kDa. This result implies the possible presence of a cleavage site in the NS5A sequence around aa 150, which is exposed through conformational alteration upon the N-terminal deletions. PMID- 11018281 TI - Rat cytomegalovirus R89 is a highly conserved gene which expresses a spliced transcript. AB - In all sequenced herpesvirus genomes, a homolog of the herpes simplex virus type 1 UL15 gene has been identified. This gene encodes a protein that is involved in viral genome maturation. Although transcription of the alphaherpesvirus UL15 gene has been analyzed in detail, not much is known about the expression of its betaherpesvirus homologs. We therefore set out to characterize transcription of the rat cytomegalovirus counterpart of UL15, R89. Here we report that R89 consists of two exons separated by a 4.7-kb intron. The spliced R89 transcript, which is expressed at late times postinfection (p.i.), has the capacity to encode a protein of 670 amino acids with a calculated molecular mass of 77.1 kDa. The predicted amino acid sequence of this protein is highly similar to that of the proteins predicted to be encoded by the human cytomegalovirus UL89 and murine cytomegalovirus M89 genes (64.3 and 84.5% overall identity, respectively). The region between R89 exon 1 and exon 2 was found to contain five additional genes, r90, R91, R92, R93 and R94, the latter two of which are conserved among all herpesviruses. We show that these genes are transcribed in a highly complex fashion, resulting in numerous mono- and polycistronic mRNAs. PMID- 11018282 TI - The complete nucleotide sequence and synthesis of infectious RNA of genomic and defective interfering RNAs of TBSV-P. AB - The complete nucleotide sequences of the genome of the pepper isolate of tomato bushy stunt Tombusvirus (TBSV-P), and its defective interfering (DI) RNAs were determined. The genome of TBSV-P is a linear single-stranded monopartite RNA molecule of positive polarity, 4776 nucleotides long and has an organisation identical to that reported for other tombusviruses. In vitro transcripts of the genome were highly infectious, and it could support replication of the DI RNAs associated with the wild type virus. Two DI RNAs were found in the infected leaves of Nicotiana clevelandii, whose sequences were completely derived from the genomic RNA. The longest DI RNA (DI-5) has 550 nucleotides (nt), while the shorter DI RNA (DI-4) composed of 463 nt, both of them were formed by essentially the same genomic sequence blocks. Since host specificity of TBSV-P and other tombusviruses with available infectious cDNA clones is different, it is feasible to carry out gene exchange studies to determine viral host specificity factors for tombusviruses. PMID- 11018283 TI - Mutations in conserved regions of ribosomal RNAs decrease the productive association of peptide-chain release factors with the ribosome during translation termination. AB - Early studies provided evidence that peptide-chain release factors (RFs) bind to both ribosomal subunits and trigger translation termination. Although many ribosomal proteins have been implicated in termination, very few data present direct biochemical evidence for the involvement of rRNA. Particularly absent is direct evidence for a role of a large subunit rRNA in RF binding. Previously we demonstrated in vitro that mutations in Escherichia coli rRNAs, known to cause nonsense codon readthrough in vivo, reduce the efficiency of RF2-driven catalysis of peptidyl-tRNA hydrolysis. This reduction was consistent with the idea that in vivo defective termination at the mutant ribosomes contributes to the readthrough. Nevertheless, other explanations were also possible, because still missing was essential biochemical evidence for that idea, namely, decrease in productive association of RFs with the mutant ribosomes. Here we present such evidence using a new realistic in vitro termination assay. This study directly supports in vivo involvement in termination of conserved rRNA regions that also participate in other translational events. Furthermore, this study provides the first strong evidence for involvement of large subunit rRNA in RF binding, indicating that the same rRNA region interacts with factors that determine both elongation and termination of translation. PMID- 11018284 TI - A novel mutation in ribosomal protein S4 that affects the function of a mutated RF1. AB - Release factors (RF) 1 and 2 trigger the hydrolysis of the peptide from the peptidyl-tRNA during translation termination. RF1 binds to the ribosome in response to the stop codons UAG and UAA, whereas RF2 recognizes UAA and UGA. RF1 and RF2 have been shown to bind to several ribosomal proteins. To study this interaction in vivo, prfA1, a mutant form of RF1 has been used. A strain with the prfA1 mutation is temperature sensitive (Ts) for growth at 42 degrees C and shows an increased misreading of UAG and UAA. In this work we show that a point mutation in ribosomal protein S4 can, on the one hand, make the RF1 mutant strain Ts(+); on the other hand, this mutation increases the misreading of UAG, but not UAA, caused by prfA1. The S4 mutant allele, rpsD101, is a missense mutation (Tyr51 to Asp), which makes the cell cold sensitive. The behaviour of rpsD101 was compared to the well-studied S4 alleles rpsD12, rpsD14, and rpsD16. These three mutations all confer both a Ts (44 degrees C) phenotype and show a ribosomal ambiguity phenotype, which rpsD101 does not. The three alleles were sequenced and shown to be truncations of the S4 protein. None of the three mutations could compensate for the Ts phenotype caused by the prfA1 mutation. Hence, rpsD101 differs in all studied characteristics from the three above mentioned S4 mutants. Because rpsD101 can compensate for the Ts phenotype caused by prfA1 but enhances the misreading of UAG and not UAA, we suggest that S4 influences the interaction of RF1 with the decoding center of the ribosome and that the Ts phenotype is not a consequence of increased readthrough. PMID- 11018285 TI - Overproduction and improved strategies to purify the threenative forms of nuclease-free HU protein. AB - The heterodimeric HU protein was isolated from Escherichia coli as one of the most abundant DNA binding proteins associated with the bacterial nucleoid. HUalphabeta is composed of two very homologous subunits, but HU can also be present in E. coli under its two homodimeric forms, HUalpha(2) and HUbeta(2). This protein is conserved either in its heterodimeric form or in one of its homodimeric forms in all bacteria, in plant chloroplasts and in some viruses. HU can participate, like the histones, in the maintenance of DNA supercoiling and in DNA condensation. This protein which does not recognize any specific sequence on double-stranded DNA, has been shown to bind specifically to cruciform DNA as does the eukaryotic HMG1 protein and to a series of structures which are found as intermediates of DNA repair, e.g., nick, gap, 3'overhang, etc. The strong binding of HU to these diverse DNA structures could explain, in part at least, its pleiotropic role in the bacterial cell. To understand all the facets of its interactions with nucleic acids, it was necessary to develop a procedure which allowed the purification of the three forms of HU under their native form and without the nuclease activity strongly associated with the protein. We describe here such a procedure as well as demonstrating that the three histidine-tagged HUs we have produced, have conserved the binding characteristics of native HUs. Interestingly, by two complementation tests, we show that the histidine-tagged HUs are fully active in vivo. PMID- 11018286 TI - Myristoylation-dependent N-terminal cleavage of the myristoylated alanine-rich C kinase substrate (MARCKS) by cellular extracts. AB - The myristoylated alanine-rich C kinase substrate (MARCKS) has been proposed to regulate the plasticity of the actin cytoskeleton at its site of attachment to membranes. In macrophages, MARCKS is implicated in various cellular events including motility, adhesion and phagocytosis. In this report we show that macrophage extracts contain a protease which specifically cleaves human MARCKS, expressed in a cell-free system or in E. coli, between Lys-6 and Thr-7. Cleavage of MARCKS decreases its affinity for macrophage membranes by ca. one order of magnitude, highlighting the contribution of the myristoyl moiety of MARCKS to membrane binding. Importantly, cleavage requires myristoylation of MARCKS. Furthermore, MARCKS-related protein (MRP), the second member of the MARCKS family, is not digested. Since Thr-7 is lacking in MRP this suggests that Thr-7 at the P1 position is important for the recognition of lipid-modified substrates. A different product is observed when MARCKS is incubated with a calf brain cytosolic extract. This product can be remyristoylated in the presence of myristoyl-CoA and N-myristoyl transferase, demonstrating that cycles of myristoylation/demyristoylation of MARCKS can be achieved in vitro. Although the physiological relevance of these enzymes still needs to be demonstrated, our results reveal the presence of a new class of cleaving enzymes recognizing lipid modified protein substrates. PMID- 11018287 TI - Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA. AB - The presence of apurinic/apyrimidinic (AP) sites in cell genomes is known to be toxic and mutagenic. These lesions are therefore repaired in cells by efficient enzymatic systems. However, a report (Nakamura and Swenberg, Cancer Res. 59 (1999) 2522-2526) indicates an unexpected high rate of endogenous apurinic/apyrimidinic (AP) sites in genomic DNA in mammalian tissues. The technology used does not allow the authors to distinguish between intact AP sites and 3'cleaved AP sites. The corresponding values range between 2 and 4 sites per million of nucleotides in various human and rat tissues. Using a modified alkaline elution method we show here that the stationary level of intact AP sites is about 0.16 per million of nucleotides in leukemic mouse L1210 cells. PMID- 11018288 TI - Peroxisome proliferator-activated receptor-alpha expression in rat liver during postnatal development. AB - The expression of the peroxisome proliferator-activated receptor-alpha (PPARalpha) as well as of some related genes was studied in rat liver at different stages of development (from 19-day-old fetuses to 1 month-old rats). The level of PPARalpha mRNA appeared higher in neonates than in fetuses or 1 month-old rats. Whereas the pattern for phosphoenolpyruvate carboxykinase (PEPCK) mRNA level was similar to that of PPARalpha, the mRNA level of both acyl-CoA oxidase (ACO) and apolipoprotein CIII (apo CIII) showed diverse profiles. Western blotting analysis also revealed an increased level of PPARalpha protein in liver of suckling rats. Similarities of mRNA PEPCK and PPARalpha expression indicate a common control mechanism, where both nutritional and hormonal factors may be involved. PMID- 11018289 TI - C-terminal region of the cytosolic subunit p47(phox) is a primary target of conformational change during the activation of leukocyte NADPH oxidase. AB - The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of O(2(-)) from oxygen using NADPH as the electron donor. During activation, the cytosolic oxidase components p47(phox) and p67(phox), each containing two Src homology 3 (SH3) domains, migrate to the plasma membrane, where they associate with cytochrome b(558), a membrane integrated flavohemoprotein, to assemble the active oxidase. Oxidase activation can be mimicked in a cell-free system using an anionic amphiphile, such as sodium dodecyl sulfate or arachidonic acid and the phosphorylation of p47(phox )with protein kinase C. Activators of the oxidase in vitro cause exposure of p47(phox) SH3, which has probably been masked by the C-terminal region of this protein in a resting state. We show here that the total protein steady-state intrinsic fluorescence exhibited by the tryptophan residues of p47(phox) substantially decreased when N-terminal truncated p47(phox)-SH3-C was treated with anionic amphiphiles or phosphorylated with protein kinase C. This finding was similar to the results obtained with full-length p47(phox). However, the fluorescence of C terminal truncated p47(phox)-N-SH3 and both C-terminal and N-terminal truncated p47(phox)-SH3 were not altered by the activators. These results indicate that the C-terminal region of p47(phox) is a primary target of the conformational change during the activation of NADPH oxidase. PMID- 11018290 TI - The role of ERG20 gene (encoding yeast farnesyl diphosphate synthase) mutation in long dolichol formation. Molecular modeling of FPP synthase. AB - The yeast Saccharomyces cerevisiae strain LB332 bearing a mutation in the ERG20 gene encoding farnesyl diphosphate synthase (FPPS) synthesizes significantly longer dolichols than the wild type strain FL100 (14-31 and 14-19 isoprene units, respectively). The measurement of the short chain prenyl alcohols excreted into the medium shows that increased amounts of geraniol, dimethylallyl and isopentenyl alcohols but not farnesol are synthesized by the mutant strain. The wild type FPPS synthesizes farnesyl diphosphate (FPP) as the only product. The K197E substitution, as opposed to F112A/F113S in avian FPPS, does not change product specificity. Consequently, the possibility that mutated yeast FPPS synthesizes longer polyprenols is unlikely. This is supported by additional evidence such as in vitro analysis of the mutated FPPS products and molecular modeling. We suggest that formation of longer dolichols in vivo is the result of a change in the isopentenyl diphosphate/farnesyl diphosphate ratio caused by the erg20 mutation which in turn affects the activity of cis-prenyltransferase. PMID- 11018291 TI - Solution structure of the orphan nuclear receptor rev-erb beta response element by 1H, 31P NMR and molecular simulation*. AB - Rev-erb beta is an orphan receptor that binds as a homodimer or as a monomer to DNA. The solution structure of the non-palindromic 15 bp DNA duplex d(TAGAATGTAGGTCAG), the response element of Rev-erb beta for monomeric binding, was determined by 1H and 31P NMR, energy minimization with NMR-derived restraints for distances and NOE back-calculation methods. The refined final structures have the typical overall features of B-type DNA. However, titration of this 15 bp duplex with ReDBD, the DNA binding domain of Rev-erb beta, showed large shifts of imino protons and 31P signals, suggesting major conformational changes. PMID- 11018292 TI - Ciprofibrate stimulates protein kinase C-dependent phosphorylation of an 85 kDa protein in rat Fao hepatic derived cells. AB - The effect of ciprofibrate on early events of signal transduction was previously studied in Fao cells. Protein kinase C (PKC) assays performed on permeabilized cells showed a more than two-fold increase in PKC activity in cells treated for 24 h with 500 microM ciprofibrate. To show the subsequent effect of this increase on protein phosphorylation, the in vitro phosphorylation on particulate fractions obtained from Fao cells was studied. Among several modifications, the phosphorylation of protein(s) with an apparent molecular mass of 85 kDa was investigated. This modification appeared in the first 24 h of treatment with 500 microM ciprofibrate. It was shown to occur on Ser/Thr residue(s). It was calcium but not calmodulin-dependent. The phosphorylation level of this/these protein(s) was reduced with kinase inhibitors and especially with 300 nM GF-109203X, a specific inhibitor of PKC. All these results suggest that the phosphorylation of the 85 kDa protein(s) is due to a PKC or to another Ser/Thr kinase activated via a PKC pathway. A possible biochemical candidate for 85 kDa protein seems to be the beta isoform of phosphatidylinositol 3-kinase regulatory subunit. PMID- 11018293 TI - Myotoxic phospholipases A(2) in bothrops snake venoms: effect of chemical modifications on the enzymatic and pharmacological properties of bothropstoxins from Bothrops jararacussu. AB - Venoms from eight Bothrops spp. were fractionated by ion-exchange chromatography on CM-Sepharose at pH 8.0 for the purification of myotoxins. Chromatographic profiles showed differences regarding myotoxic components among these venoms. B. alternatus, B. atrox and B. jararaca venoms did not show the major basic myotoxic fractions identified in the other venoms. Polyacrylamide gel electrophoresis for basic proteins also showed distinct patterns for these toxins. In vivo, all the isolated myotoxins induced release of creatine kinase due to necrosis of muscle fibers, accompanied by polymorphonuclear cell infiltration, and edema in the mouse paw. In addition, the toxins showed cytotoxic and liposome-disrupting activities in vitro. B. jararacussu bothropstoxins-I (BthTX-I) and II (BthTX-II) were submitted to chemical modifications of: His, by 4-bromophenacyl bromide (BPB) or photooxidation by Rose Bengal (RB); Tyr, by 2-nitrobenzenesulphonyl fluoride (NBSF); and Trp, by o-nitrophenylsulphenyl chloride (NPSC). The myotoxic and cytotoxic activities of BthTX-I, a Lys49 PLA(2) homologue, after modification by BPB, RB, NBSF and NPSC, were reduced to 50%, 20%, 75%, 65% and 13%, 0.5%, 76%, 58%, respectively. However, the edema-inducing and liposome-disrupting activities were not significantly reduced by the above modifications. BPB-treated BthTX-II, an Asp49 PLA(2) homologue, lost most of its catalytic, indirect hemolytic, anticoagulant, myotoxic and cytotoxic activities. The edema-inducing and liposome disrupting activities were reduced to 50% and 80%, respectively. Lethality caused by BthTX-I and -II was strongly reduced after treatment with BPB or RB, but only partially with NBSF or NPSC. BthTX-I and -II, both native or modified, migrated similarly in a charge-shift electrophoresis. Antibodies raised against BthTX-I or -II, B. asper Basp-II and the C-terminal 115-129 peptide from Basp-II did not show significant differences in their cross-reactivity with the modified toxins, except with RB photooxidized toxins. PMID- 11018294 TI - Proteolytic fragmentation of polypeptide release factor 1 of Thermus thermophilus and crystallization of the stable fragments. AB - Polypeptide release factor one from Thermus thermophilus, ttRF1, was purified and subjected to crystallization. Thin crystalline needles were obtained but their quality was not satisfactory for X-ray diffraction. Stable fragments of ttRF1 suitable for crystallization were screened by limited proteolysis. Three major fragments were produced by thermolysinolysis and analyzed by N-terminal sequencing and electrospray mass spectrometry. They were N-terminal fragments generated by proteolysis at amino acid positions 211, 231 and 292. The corresponding recombinant polypeptides, ttRF1(211), ttRF1(231) and ttRF1(292), were overproduced and subjected to crystallization. Of these polypeptides, ttRF1(292) gave rise to crystals that belong to P3(1) (or P3(2)) space group with unit cell parameters a = b = 64. 5 A, c = 86.6 A and diffract up to 7 A resolution. PMID- 11018295 TI - Polydispersity and heterogeneity of squid cranial cartilage proteoglycans as assessed by immunochemical methods and electron microscopy. AB - The three populations of squid cranial cartilage proteoglycans, D1D1A, D1D1B and D1D2 appeared to have a high degree of polydispersity. Gel electrophoresis and immunoblotting analysis showed that polydispersity was mainly due to the variable size of chondroitin sulphate E chains. This was further ascertained after rotary shadowing electron microscopy of proteoglycan core proteins and glycosaminoglycan side chains and statistical analysis of the sizes measured for both components. Enzymic treatment of the proteoglycan core proteins produced different peptides from each population, suggesting that the observed heterogeneity of the proteoglycans is due to their core proteins. Antibodies were raised in rabbits against all proteoglycans and enzyme-linked immunosorbent analysis of proteoglycan core proteins revealed that the proteoglycans, even heterogeneous, shared many common epitopes. Part of the common proteoglycan epitopes were found to be located in chondroitin sulphate E chains. Heterogeneity of squid proteoglycans was also investigated by studying their interactions with collagen and it was found that only the two populations of high molecular mass, D1D1A and D1D2, were able to interact with only collagen type I, the latter stronger than the former. PMID- 11018296 TI - Prediction of protein structural classes by neural network. PMID- 11018297 TI - Prenatal melatonin exposure affects luteinizing hormone and hypothalamic and striatal neuropeptide Y in the male rat offspring. AB - The present study examines the influence of prenatal melatonin on the hypothalamic and striatal neuropeptide Y (NPY) concentrations as well as on luteinizing hormone (LH) levels. Male rat offspring of control and melatonin treated mother rats were studied at different ages of the sexual development: infantile, prepubertal, pubertal and adult ages. Hypothalamic NPY levels were much higher during the juvenile than throughout the infantile period. After prenatal melatonin treatment significantly higher values since day 15 up to 35, also at 60 days of age were found, as compared with controls. Striatal NPY levels were lower than in hypothalamus. Again, NPY in the striatum from offspring of melatonin treated mother rats showed significantly higher values than the respective controls at most of the ages studied. However, prenatal melatonin exerted an inhibitory influence upon LH secretion pattern, since decreased concentrations up to 25 days of age and delayed peak values at pubertal age were observed. The present study also suggest that the effect of NPY upon LH secretion is related to sexual development, since NPY exerted opposite effect in infantile than in pubertal period and melatonin administration during intrauterine life prevented this effect. PMID- 11018298 TI - Synergy between 5' and 3' flanking regions of the human tyrosine hydroxylase gene ensures specific, high-level expression in neuroblastoma cells. AB - Factors regulating tyrosine hydroxylase (TH) gene transcription are of major importance in the studies of malignant and degenerative diseases of catecholamine synthesizing tissues. In this study, we used transient transfection of a reporter gene to show that high-level, tissue-specific TH expression was only achieved when the reporter gene was cloned between a 5' TH promoter sequence (-513-+1), and, a 3' TH gene flanking sequence (end of exon 14-+976). We also show that TH mRNA expression level is closely linked to the expression level of the proto oncogene, MYCN in neuroblastoma tumor cell lines. Taken together our data indicate that MYCN may regulate TH expression in neuroblastoma cells, but not through binding to the 5' or 3' TH gene flanking sequences used in our experiments. PMID- 11018299 TI - Chronological changes of re-closure capacity in surgically induced spinal open neural tube defects of chick embryos. AB - To investigate the re-closure capacity of surgically induced spinal open neural tube defects (ONTD) and its correlation to the cell proliferation activity in embryonic stages, longitudinal incisions of 3- or 6-somite lengths were made in neural tubes of chick embryos at Hamburger and Hamilton stage 18-19, and the length of the ONTDs was measured on postoperative days 2, 3, 5, 7, and 10. The results demonstrated that re-closure occurred more actively when the incision length was shorter. The chronological changes in the re-closure capacity of the ONTDs paralleled the proliferative activity of the neural tube. We conclude that surgically induced spinal ONTDs may be remodeled in early chick embryos and that manipulation of cell dynamics can enhance the re-closure process in the embryonic spinal cord tissue. PMID- 11018300 TI - Impaired neurotransmitter systems by Abeta amyloidosis in APPsw transgenic mice overexpressing amyloid beta protein precursor. AB - APPsw transgenic mice showing substantial features of brain Abeta amyloidosis such as senile plaques and behavioral abnormalities were examined by immunostaining to determine whether Abeta deposits could induce the subsequent disturbance of neurotransmitter systems including somatostatin, substance P and choline acetyltransferase (ChAT), which are prominent in the Alzheimer's disease brain. Somatostatin, substance P and ChAT disappeared in the areas of senile plaque and were accumulated in dystrophic neurites around the amyloid cores. These findings suggest a potential role of brain Abeta amyloidosis in disturbance of the neurotransmitter systems leading to memory disturbance of Alzheimer's disease. PMID- 11018301 TI - Activation of microglia in kainic acid induced rat retinal apoptosis. AB - We applied a variety of methods to follow the course of kainic acid (KA) induced retinal apoptosis, especially with regard to the spatial and temporal aspects. At 24 h after KA injection, a massive cell increase, which showed terminal transferase-mediated dUTP nick-end-labeling technique positive signals, was observed in all of the retinal layers, with the exception of the outer nuclear and photoreceptor layers. Electron microscopy further confirmed that these cells might be apoptotic body ingesting phagocytes, whose function seemed to correlate with bcl-2 mRNA up-regulation. When histochemical studies were performed to determine the cellular identity of the phagocytes, the microglia were thought to be the one and only type of phagocytes involved in the KA-induced retinal apoptosis. In conclusion, we demonstrated that after KA injection, microglia were the only phagocytes to participate in clearing apoptotic debris from the inner retinal layers, and that their function might correlate with the change in expression of the bcl-2 gene family. PMID- 11018302 TI - Enzyme- and immunohistochemical localization of nitric oxide synthase in nerves of the porcine pancreas. AB - The distribution of nitric oxide synthase (NOS) in neuronal and non-neuronal porcine pancreatic tissues was examined using nicotinamide adenine dinucleotide hydrogen phosphate diaphorase (NADPHd) enzymehistochemistry and neuronal type NOS immunohistochemistry. NOS-containing perikarya were regularly discernible in the pancreatic ganglia, whereas positive thin nerve fibers appeared within nerve bundles of the interlobular spaces and as thin varicose fibers in the vicinity of pancreatic blood vessels. Vascular endothelium showed intense staining for NADPHd and most of the islet cells were NOS immunoreactive. The above morphological findings indicate that nitrergic innervation of the porcine pancreas is very similar to that of the pancreata of other mammalian species and suggest that nitric oxide of both neuronal and extraneuronal origin may regulate blood flow and secretion of the porcine pancreas. PMID- 11018303 TI - Expression of DNA-dependent protein kinase catalytic subunit and Ku80 in developing human brains: implication of DNA-repair in neurogenesis. AB - We investigated expression of DNA-dependent protein kinase catalytic subunit (DNA PKcs) and Ku80 in developing human brain, both of which have been suggested to be involved in the repair of DNA double-strand break (DSB). Their expressions were well correlated, and the highest immunoreactivity was observed in post-mitotic immature neurons in the cerebral cortex as well as in progenitors in the periventricular germinal layer. The reactivity gradually decreased during development. Our results support the notion that DSB is generated during the ontogeny of the human central nervous system. PMID- 11018304 TI - Suppression of experimental allergic encephalomyelitis is specific to the frequency and intensity of nocturnally applied, intermittent magnetic fields in rats. AB - Female Lewis rats (n=72) were inoculated with an emulsion of spinal cord and complete Freund's adjuvant. They were then exposed for approximately 6 min every hour between midnight and 08:00 h for 2 weeks to either 7 or 40 Hz amplitude modulated magnetic fields whose temporal pattern was designed to simulate a (geomagnetic) storm sudden commencement. The peak strengths of the fields averaged between either 30-50 nT (low intensity) or 500 nT (high intensity). Rats exposed to the 7 Hz, low intensity magnetic fields displayed significantly less severe overt signs of experimental allergic encephalomyelitis than rats exposed to either of the two intensities of the 40 Hz fields, the high intensity 7 Hz field, or the reference (<10 nT) condition. The latter groups did not differ significantly from each other. Predicted severity based upon the numbers of foci of infiltrations of lymphocytes within the brains of the rats also demonstrated the ameliorating effects of the low intensity, 7 Hz exposures. These results suggest very specific characteristics of complex, weak magnetic fields within the sleeping environment could affect the symptoms of autoimmunity. PMID- 11018305 TI - Electroencephalogram alpha (8-15 Hz) responses to visual stimuli in cat cortex, thalamus, and hippocampus: a distributed alpha network? AB - To investigate possible functional correlates of alpha (8-15 Hz) oscillations in the electroencephalogram (EEG) intracranial recordings in cats (from thalamus, occipital cortex, and hippocampus) were performed. In response to visual stimuli, event-related alpha oscillations were observed. Such alpha responses were found not only in a specific sensory (visual) pathway but also in the hippocampus, hinting at a possible distributed alpha response system. PMID- 11018306 TI - Vitamin E decreases valproic acid induced neural tube defects in mice. AB - The present study was undertaken to investigate the effect of vitamin E on valproic acid (VPA) induced teratogenesis. Pregnant Balb mice were divided into six groups of 10-11 animals each. The mice in group 1 served as control and were injected with saline subcutaneously on day 8 of gestation, whereas, animals in group 2 received a single injection of VPA (700 mg/kg (s.c.)). Groups 3 and 4 received an oral administration of vitamin E in the doses of 250 and 500 mg/kg, respectively, 1 h before VPA injection. Group 5 and 6 were given vitamin E only, in the same doses as group 3 and 4. On day 18 of gestation, the mice were killed by cervical dislocation. Embryotoxicity was assessed by counting the number of implants, live and dead fetuses, resorptions, crown rump length and fetal body weight. The fetuses were observed for malformations including neural tube defects (excencephaly), open eye lid and micrognathae. VPA administration resulted in a significant reduction of the average live fetuses/litter, fetal weight and crown rump length and a significant increase in malformations (excencephaly, open eye lid and micrognathae). Concomitant administration of vitamin E significantly attenuated VPA induced decrease in the fetal weight, crown rump length and malformations. PMID- 11018307 TI - Stimulatory and inhibitory effects of adenosine A(2A) receptors on nerve growth factor-induced phosphorylation of extracellular regulated kinases 1/2 in PC12 cells. AB - Effects of nerve growth factor (NGF), adenosine and an adenosine A(2A) receptor agonist (CGS 21680) on the phosphorylation of extracellular-regulated kinases 1/2 (ERK1/2) were examined in PC12 cells. Adenosine and CGS 21680stimulated ERK1/2, but inhibited the phosphorylation of ERK1/2 induced by a 10 min incubation with NGF. Longer treatment with CGS 21680 and NGF (1-2h) resulted in an additive effect on the activation of ERK1/2. Forskolin exerted the same effects, suggesting that they are mediated by cyclic AMP. These results indicate that adenosine A(2A) receptor induced increases in cyclic AMP can stimulate ERK1/2 phosphorylation per se, inhibit the initial and enhance the late NGF-induced activation of ERK1/2. These results may be explained by the fact that NGF action is mediated via different pathways at early and late time points. PMID- 11018308 TI - Selective nicotinamide adenine dinucleotide phosphate-diaphorase histochemical labeling of Muller radial processes and photoreceptors in the earliest stages of retinal development in the tadpole. AB - To investigate potential sources of nitric oxide production in the early stages of retinal development we used, in the tadpole, nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry that was reported to reveal nitric oxide synthase isoforms in the retina. In the first stages of optic vesicle differentiation, prior to optic cup invagination, histochemical positivity was detected in the radial processes of Muller cells, that provide a scaffold for migrating retinal neuroblasts, and was soon followed by intense staining of photoreceptors. These events preceded retinal laminar patterning and the appearance of histochemical positivity in other retinal cell populations. The findings indicate that nitric oxide synthase is expressed during early retinogenesis at selective sites, which are implicated in the guidance of migrating cells and in phototransduction. PMID- 11018309 TI - Skin sympathetic neuroeffector response is attenuated dose-dependently by systemic prostaglandin E1 injection in humans. AB - To clarify the effects of prostaglandin E1 (PGE1) on the vasoconstrictive responses, we compared the correlation between the amplitude of integrated skin sympathetic nerve activity (SSNA) and per cent reduction in skin blood flow (SBF) before and after the infusion of lipo PGE1 and placebo (bolus one-shot infusion, single blind study), and constant rate infusion of PGE1 (10 and 50 ng kg(-1) min( 1) by infusion pump, dose-dependency study) in ten healthy men. SSNA was recorded microneurographically from the median nerve simultaneously with SBF by laser Doppler flowmetry at the index fingertip. The measurement was conducted 30 min after injection of lipo PGE1 or placebo, and during the drip infusion of 10 and 50 ng kg(-1) min(-1) of PGE1 with maneuvers to enhance SSNA. The resting and activated skin blood flow were not significantly different between 10 ng lipo PGE1 and placebo administration, and between baseline and 10, 50 ng kg(-1) min( 1) of PGE1 injection. The vascular response, defined as the slope of regression line between logarithm of amplitude of integrated SSNA bursts and the reduction in SBF, was significantly suppressed by injection of lipo PGE1 as compared with that by placebo. It was also decreased dose-dependently by the constant rate infusion of PGE1 (10 and 50 ng kg(-1) min(-1)). We concluded that the intravenous injection of PGE1 attenuates vasoconstrictive responses to SSNA, and analysis of the relations between SSNA and vasoconstrictive response, i.e. the neuroeffector response, is suggested to be an important tool to assess the drug effect. PMID- 11018310 TI - Cardiovascular risk factors and Alzheimer's disease: a genetic association study in a population aged 85 or over. AB - Increasing evidence suggests a relation between vascular disorders and late-onset Alzheimer's disease (AD). We performed an association analysis of low-density lipoprotein receptor-related protein (LRP), lipoprotein lipase (LPL), and angiotensin converting enzyme (ACE) genes, known to be involved in vascular disorders, and AD. Genotyping was carried out in 113 patients with clinically defined Alzheimer's disease (NINCDS-ADRDA criteria) and 203 non-demented controls in a prospective, population-based study of people aged 85 years or over (Vantaa 85+ Study). Corresponding analysis was performed on 121 neuropathologically verified AD patients (CERAD criteria) and 75 controls derived from the same study population. We did not find significant associations between the polymorphisms studied and AD. However, analysis of the LPL polymorphism showed a weak trend (uncorrected P-value 0.095) towards protection against neuropathologically defined AD. Our study is based on very elderly Finns. Therefore, further studies are warranted in other populations. PMID- 11018312 TI - Tumor necrosis factor-alpha modulates the proliferation of neural progenitors in the subventricular/ventricular zone of adult rat brain. AB - Little is known about the response of neural progenitors to inflammation following injuries of the central nervous system. In combination with bromodeoxyuridine (BrdU) intraperitoneally (i.p.) injected, tumor necrosis factor alpha (TNF-alpha), a proinflammatory cytokine that increased ED1+ activated microglia/macrophage population at injured sites, was administrated into adult rat brains. No difference in the immunostaining for proliferating cell nuclear antigen (PCNA) was observed in the subventricular/ventricular zone (SVZ/VZ) between TNF-alpha injected sites and controls. However, BrdU+ cells were apparently observed in the SVZ/VZ proximal to TNF-alpha injected site, and the number of BrdU+ cells increased at 6 and 24 h post injection. Since cell apoptosis was rarely found in the SVZ/VZ after TNF-alpha injection, these observations suggest that the diffusible TNF-alpha may directly and/or indirectly modulate the proliferation of neural progenitors. PMID- 11018311 TI - An association study of bipolar mood disorder (type I) with the 5-HTTLPR serotonin transporter polymorphism in a human population isolate from Colombia. AB - The short variant of a functional length polymorphism in the promoter region of the serotonin transporter has been associated with several behavioural and psychiatric traits, including bipolar mood disorder. The same short allele has also been implicated as a modifier of the bipolar phenotype. Here we evaluate the etiologic/modifier role of this polymorphism in a case (N=103) / control (N=112) sample for bipolar mood disorder (type I) collected from an isolated South American population. We did not detect an association between bipolar disorder and the 5-HTT promoter polymorphism in this sample. However, an excess of the short allele was seen in younger cases and in cases with psychotic symptoms. When combined with data from the literature, the increased frequency of the short allele in patients with psychotic symptoms was statistically significant. PMID- 11018313 TI - Interleukin-1beta reduces temperature sensitivity but elevates thermal thresholds in different populations of warm-sensitive hypothalamic neurons in rat brain slices. AB - Extracellularly recorded firing rates of neurons in slices of the preoptic area and anterior hypothalamus (PO/AH) of the rat were determined during thermal stimulation. Human recombinant interleukin-1beta (20 ng/ml) did not influence temperature-insensitive neurons, but reduced the firing rate and thermosensitivity in linear warm-sensitive neurons, and shifted the thermal thresholds of activation in threshold warm- and cold-sensitive neurons by 1.1-2.3 degrees C to hyperthermic temperatures. The data support the suggestion that endogenous pyrogens may act on different populations of thermosensitive PO/AH neurons to induce fever. The shift of the thermal thresholds of activation of threshold warm- and cold-sensitive neurons in combination with the otherwise maintained temperature sensitivity of these neurons appears to play a major part for the controlled shift of body temperature and the maintenance of the elevated body temperature during cytokine-induced fever. PMID- 11018314 TI - Brain oscillations control hand orthosis in a tetraplegic. AB - The aim of the present study was to investigate whether self-induced brain potential changes could be useful as control signals for patients with severe motor impairment, i.e. due to high-level spinal cord injury. The pilot project was performed in a tetraplegic patient (T.S.), whose residual muscle activity of the upper limbs is restricted to the left biceps. To restore the hand grasp function, an electrical driven hand orthosis fitting his left hand was developed. The operation of this device is directly based on the bioelectrical signals of the brain. After some months of training, T. S. has learned to operate the hand orthosis by mental imagination of specific motor commands. PMID- 11018315 TI - Temporal stream of cortical representation for auditory spatial localization in human hemispheres. AB - We measured human evoked magnetic fields to binaural sounds with an interaural time delay as a cue for auditory localization. By analyzing the topography of auditory-evoked magnetic fields in the middle-latency, we demonstrated that particular cortical regions represent the direction of sound localization by their activity level. Upon presenting a binaural sound, the first representations were found in the middle frontal region as well as the superior temporal region of the right hemisphere approximately 19 ms after the stimulation, but their patterns differed. Other cortical regions including the prefrontal and parietal spatial areas were affected within roughly 60 ms. The results showed that the right hemisphere is dominant even in the preattentive stage of auditory spatial processing of sounds from different directions. PMID- 11018316 TI - Immunocytochemical localization of neuronal and inducible nitric oxide synthase in the retina of zebrafish, Brachydanio rerio. AB - Several previous studies have revealed the distribution of neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) in the retina of various species. However, nothing has been reported on the nNOS and iNOS expression in zebrafish retina, even though it has been used as an effective model for developmental studies. In this study on nNOS and iNOS immunoreactivity (IR) in the zebrafish retina, iNOS-IR signals were detected in the ganglion cell layer (GC), the inner plexiform layer (IP), the outer plexiform layer (OP) and the photoreceptor layer (PR). nNOS-IR signals were also intensely detected in the GC, IP, OP and PR. These signals were very similar to the iNOS-IR signals, except for some minor differences in relative intensities. This could be explained by the speculation that the synthesis of nitric oxide by iNOS may represent a compensatory mechanism in the absence of nNOS. We described for the first time the distribution of nNOS and iNOS-IR cells in zebrafish retina. This could provide the basis for further study on the nNOS and iNOS properties of zebrafish retina. PMID- 11018334 TI - Palliative care consultations: how do they impact the care of hospitalized patients? AB - To provide a detailed description of the recommendations of a Palliative Care Service (PCS) and to describe the impact of these recommendations on the care of terminally ill patients in an academic medical center, we describe data from all consecutive patients referred by their attending physicians to the PCS of an academic teaching hospital over a 15-month period. All patients were seen within 24 hours of consultation request. Data were collected prospectively on the day of discharge or death. Attention was focused on six recommendations and their implementation: 1) discussion about prognosis and goals of care; 2) pursuing documentation of advance directives; 3) discussion about foregoing specific treatments and/or diagnostic interventions; 4) family and patient support; 5) discharge planning; and 6) symptom management. Over a 15-month period, we collected data on 325 patients. The most frequent diagnoses were cancer, dementia, and HIV disease. The patients were followed for a mean of 7.6 days. The average number of recommendations was 4.2 per patient and 91% of the recommendations were implemented (3.8 per patient). Recommendations increased to 5.3 per patient and the implementation rate increased to 97% (5.1 per patient) for the 44 patients transferred to the Palliative Care Unit (PCU). PCS consultations result in multiple recommendations with a very high implementation rate. The number of recommendations and the high implementation rate suggest a strong need for palliative care services within acute care hospitals. PMID- 11018335 TI - An international survey of undergraduate medical education in palliative medicine. AB - A 9-item mail survey dealing with availability and characteristics of undergraduate medical education programs in palliative medicine was sent to all medical schools in Canada (16) and the United Kingdom (UK) (30), and 129 randomly selected medical schools in the United States (US) and Western Europe. The overall response rate was 117/175 (67%). The highest percentage of mandatory (required by the university) rotations in palliative medicine was in the UK medical schools (14/22, 64%). Considerably lower numbers were obtained from the other countries: US; 4/37, 11%, Canada; 2/14, 14%, and Western Europe; 8/43, 19% (P = 0.001). Elective rotations in palliative medicine were more readily available in the UK; 18/22, 82% and Canada; 10/14, 71%, compared with the US; 23/37, 62%, and Western Europe; 13/43, 30% (P=0.001). Seventy-two percent (13/18) of UK, 70% (7/10) of Canadian, 59% (16/27) of US, and 9/30 (30%) of Western European medical schools provide educational reading material in palliative medicine (P = 0.014). Case-based learning in small groups and small group discussion were favored by the UK, 14/22 (63%) and 17/22 (77%), respectively, and Canadian medical schools, 8/14 (57%) and 8/14 (57%), respectively (P = 0.176). The number of universities with academic faculty positions for palliative medicine and the median number of positions for the countries were as follows Canada 8/13 (62%) and 2; UK 12/22 (55%) and 1; US 5/36 (14%) and 1; and Western Europe 9/24 (21%) and 1, respectively (P = 0.001). Besides the UK, mandatory (required) rotations in undergraduate palliative medicine education are lacking in Canadian, US, and Western European medical schools. The median number of 1 academic faculty member per responding medical school is discouraging. In order for undergraduate and postgraduate medical education in palliative medicine to improve, the number of both educational programs and faculty members will need to be increased. PMID- 11018336 TI - Psychological correlates of opioid use in patients with chronic nonmalignant pain: a preliminary test of the downhill spiral hypothesis. AB - There is still controversy surrounding the use of opioid medication for patients with chronic nonmalignant pain. Schofferman has argued that long-term opioid use leads to a "downhill spiral" associated with loss of functional capacity and a corresponding increase in depressed mood. The present study was a retrospective comparison of opioid users vs. non-users to determine whether: (a) users have higher levels of disability, medical visitation, depression, and pain; (b) the behavioral problems associated with opioid use persist after controlling for the influence of other medication; (c) opioid use is in fact a predictor of illness behavior; and (d) higher levels of opioid consumption are associated with higher levels of disability and depression. A consecutive series of 243 patients with nonmalignant pain about to enroll at a tertiary clinic were retrospectively assigned to either an Opioid User (n = 87) or Non-User (n = 156) group. Compared to Non-Users, Opioid Users were more likely to be physically disabled ( P <0.05) and depressed ( P<0.05), as well as more likely to report pain at higher levels (P<0.001) and in more locations ( P<0.05). Despite the appearance of a downhill spiral, we were unable to demonstrate an association between opioid use and any measure of illness behavior after controlling for benzodiazepine use (with the possible exception of domestic disability). Instead, we found that benzodiazepine use was significantly associated with activity level ( P<0.05), medical visitation ( P<0.01), domestic disability ( P<0.01), depression ( P <0.01), and to a lesser degree, disability days (P<0.1). Using somatization as a reference variable, we found that opioid use failed to explain a comparable amount of variance in illness behavior. Finally, there was no evidence that higher levels of opioid use were associated with higher levels of disability or depression. PMID- 11018337 TI - Assessment of pain experiences after elective surgery. AB - Pain in the postoperative period has remained a clinical problem in spite of major progress in pain assessment and management. The aim of the present study was to describe in detail the time course of pain experiences of surgical patients (n=200) following elective surgical procedures. Visual analogue scale (VAS, 0-100 mm) was used for pain intensity ratings at 4, 24, 48, and 72 hours after surgery. Interviews were carried out to assess the occurrence of intermittent worst pain episodes during each 24-hour period and to relate such experiences to clinical events. At 4, 24, 48, and 72 hours postoperatively, 39%, 43%, 27%, and 16% of the patients, respectively, experienced moderate or even severe pain (VAS > or =40 mm) at rest. During the first 24 hours after surgery, 88% of the patients had experienced moderate or severe pain at some time (VAS > or =40 mm). Corresponding figures for the following 24 hour periods were 81% and 72%, respectively. Spontaneous pain breakthrough and movement/mobilization were identified as reasons for the worst pain episodes during the first 72-hour period after surgery. The probability of pain intensity of VAS >40 mm was calculated from individual regression functions. The probability was found to be much higher for patients receiving parenteral analgesics than for patients receiving prolonged epidural analgesia. Despite major improvements in pain assessment and management, postoperative patients often experience moderate to severe pain, and worst pain episodes occur even in the late postoperative phase. The present study emphasizes that in the clinical routine management of pain, further quality assurance efforts are necessary. PMID- 11018338 TI - Interactive collaborative consultation model in end-of-life care. AB - With interactive communication becoming an engaging and accessible venue, the specialized field of end-of-life care could greatly benefit by utilizing the Internet to ensure comprehensive palliative care for many underserved areas worldwide. Interactive communication can enhance the care of patients with advanced disease and their families by providing practitioners with current research-based information to empower their clinical decision-making. A collaborative consultative Internet relationship can support practitioners in providing comprehensive palliative interventions for their patients in a timely manner. This paper describes the development of the interactive collaborative consultation model and its demonstration between a rural palliative care nurse practitioner and an urban medical research physician. Expert consultation for individual patients through the Internet can help improve access to palliative consultation and, ultimately, enhance the care of many dying persons living within underserved or remote areas worldwide. PMID- 11018339 TI - Respiratory failure due to the combined effects of transdermal fentanyl and epidural bupivacaine/diamorphine following radical nephrectomy. AB - The transdermal therapeutic system (TTS) fentanyl has been designed for rate controlled drug delivery. When the system is applied, a fentanyl depot concentrates in the upper skin layers. Plasma concentrations are not measurable until 2 hours after application, and it takes an 8-16 hr latency period until full clinical fentany effects are observed. Following removal, serum fentanyl concentrations decline gradually and fall to about 50% in approximately 16 hours. We report the case of a 77-year-old man with a history of severe arthritis, who was receiving transdermal fentanyl and developed respiratory failure after starting epidural diamorphine/bupivacaine for postoperative pain relief following radical nephrectomy. PMID- 11018340 TI - Cytokines and advanced cancer. AB - Cytokines have a major role in promoting the growth and spread of cancers. Elevated levels of several cytokines have been described in cancer patients. Evidence from animal and human studies suggests that cytokines may contribute to a wide range of symptoms in advanced cancer, including: asthenia, pain, drowsiness, cognitive failure, agitated delirium, autonomic dysfunction, anorexia, cachexia, fever and metabolic abnormalities. Considerable effort is being directed at finding anticytokine treatments, raising the possibility of new options for symptoms that are currently difficult to control. PMID- 11018341 TI - Case presentation: demise by decision, dementia, or depression? PMID- 11018342 TI - Commentary: respecting autonomy but avoiding benevolent paternalism. PMID- 11018343 TI - Is smoking associated with the risk of developing Alzheimer's disease? Results from three Canadian data sets. AB - PURPOSE: To determine whether smoking is associated with Alzheimer's disease (AD). METHODS: Analyses were conducted using three Canadian data sets: the University of Western Ontario Dementia Study (200 cases, 163 controls), the Canadian Study of Health and Aging (258 cases, 258 controls), and the patient database from the Clinic for Alzheimer Disease and Related Disorders at the Vancouver Hospital and Health Sciences Centre (566 cases, 277 controls). The association between smoking and AD was investigated using bivariate analyses and multiple logistic regression models adjusted for the potential confounders age, sex, educational level, family history of dementia, head injury, and hypertension. RESULTS: The results of bivariate analyses were inconsistent across the three data sets, with smoking status a significant protective factor, a significant risk factor, or not associated with AD. The results of multiple logistic regression models, however, were consistent: any association between smoking status and AD disappeared in all three data sets after adjustment for confounders. CONCLUSIONS: Smoking status was consistently not associated with AD across all three data sets after adjustment for confounders. Failure to adjust for relevant confounders may explain inconsistent reports of the influence of smoking on AD. Any protective effect of smoking may be limited to specific AD subtypes (e.g., early onset AD). PMID- 11018344 TI - Differences and trends in antioxidant dietary intake in smokers and non-smokers, 1980-1992: the Minnesota Heart Survey. AB - PURPOSE: Differences and secular trends in dietary antioxidant vitamin intake (vitamins E, C, and beta-carotene) in current non-smokers, light smokers, and heavy smokers were examined as part of the Minnesota Heart Survey. METHODS: Three cross-sectional surveys were conducted in adults ages 25-74 years in 1980-82 (N = 1682), 1985-87 (N = 2326), and 1990-92 (N = 2487). Dietary information was obtained from a 24-hour dietary recall. Smoking was assessed through self-report. Intakes were adjusted for age, energy intake, body mass index, education level, and exercise level (vitamins E, C and beta-carotene). RESULTS: Antioxidant vitamin intakes were significantly higher in non-smokers than in light (1-20 cig/day) and heavy smokers (>20 cig/day) when all three survey periods were combined. In men, mean vitamin E intake was 9.2 mg, 8.6 mg, and 7.8 mg for non smokers, light smokers, and heavy smokers, respectively. Results were similar in men for beta-carotene (non-smokers 1408 microg, light smokers 1287 microg, and heavy smokers 1064 microg), and vitamin C (non-smokers 81 mg, light smokers 67 mg, and heavy smokers 56 mg). Women had results of similar magnitude and direction. From 1980-92, secular trends in men showed non-significant increases from 1980-82 to 1990-92 in beta-carotene (+6.1%), while decreases were observed in vitamins E (-1.1%) and C (-2.6%). In contrast, women had large decreases in all antioxidant vitamin intakes: vitamin E (-13%), vitamin C (-18.6%), and beta carotene (-16.2%). CONCLUSIONS: Light and heavy smokers had a significantly lower overall mean dietary antioxidant vitamin intake than non-smokers. Over the decade, antioxidant dietary intake remained relatively stable in men and decreased in women in Minneapolis-St. Paul, despite improvements in access to antioxidant rich fruits and vegetables. PMID- 11018345 TI - A comparison between BMI and Conicity index on predicting coronary heart disease: the Framingham Heart Study. AB - PURPOSE: This study examined the relationship of mortality and morbidity of coronary heart disease with body mass index (BMI) and Conicity index (CI). METHODS: Among 5209 Framingham Heart Study participants, 1882 men and 2373 women had waist and weight measurement at the 4th examination period and height measured on the 5th visit. These were used for BMI and CI. RESULTS: During a 24 year follow-up, 597 men and 468 women developed CHD and 248 men and 150 women died from CHD associated causes. In men the relative risks (RR) (95% confidence interval) adjusted for age, hypertension, diabetes, smoking status, and total cholesterol for CHD incidence in 2nd, 3rd, and 4th quartiles of BMI were 1.28 (1.0, 1.65), 1.45 (1.13, 1.86), and 1.53 (1.19, 1.96). The RR for CHD incidence in the 4th quartile of BMI in women was 1.56 (1.16, 2.08). No CI quartiles were risk factors for CHD incidence. There was 86% higher risk of CHD related death in the 4th quartile of BMI than the 1st quartile of BMI in women. In men no significantly higher risks of death were found across the quartiles of BMI. No associations were found between CI quartiles and CHD mortality. CONCLUSIONS: Obesity as measured by BMI is an important risk factor for CHD incidence in men and women and for CHD mortality in women. CI was not associated with an increase in CHD incidence or mortality. Thus, BMI is a better marker than CI for predicting CHD incidence and mortality. PMID- 11018348 TI - The impact of school-organized sport activities on the priority youth health-risk behaviors. AB - PURPOSE: To evaluate the impact of school-organized sport activities on the priority youth health-risk behaviors.METHODS: Data were taken from The 1997 National School-based Youth Risk Behavior Survey. Measures of prevalence rates in six categories of priority health-risk behaviors defined by Centers for Disease Control and Prevention were performed between participants and non-participants in school-organized sport activities. Then multiple logistic regression analyses were used to assess the association between participation in school-organized sport activities and the priority health-risk behaviors.RESULTS: Overall, 49.5% of students nationwide participated in school-organized sport activities in 1997. The major behavioral factors that proved statistically significant in predicting participation in school-organized activities in the logistic regression model were non-use of seatbelt, non-use of helmet, and use of chewing tobacco. Behavioral factors that were inversely related to participation in school organized activities were smoking cigarettes, using different kinds of drugs, using condoms, carrying weapons, eating vegetables, and attempting suicide.CONCLUSIONS: Based on the nationally representative data, participation in school-organized sport activities was positively associated with many desirable health behaviors. However, a challenge for us will be not only to promote youth participation in school-organized sport activities, but also to address some co-existing behavioral problems that may lead to non-intentional injury and use of chewing tobacco. PMID- 11018349 TI - Empowering communities to remove the pump handle. An emerging role in epidemiology. AB - PURPOSE: The epidemiologist's role in relation to the populations they study needs to be reconsidered in order to maximize the contribution of epidemiology to public health, and to balance science with social responsibility.METHODS: Historical analysis and a case study of community-based, collaborative epidemiology conducted in Eastern Virginia as the basis for identifying and improving child health outcomes.RESULTS: The origins of epidemiology in Snow's work on cholera provides a dynamic model of etiologic research combined with community intervention to prevent disease. While in past decades, epidemiology was excessively objectivist and focused on identifying individual-level risk factors, epidemiologists are now reclaiming a vital role in community health promotion. Yet the historic role of the epidemiologist, whether as the white knight who saves the community from an epidemic, or alternatively, as the remote scientist who studies populations but bears no responsibility for improving their health, is no longer viable. The emerging role is that of collaborator, a scientist working in partnership with communities. In Eastern Virginia, academic epidemiologists work closely with diverse community-based coalitions to assess child health needs, prioritize health problems, design and enact interventions, and monitor health outcomes. Diagnostic studies of child health outcomes, published in peer-reviewed journals and in community reports, have identified serious health needs in Eastern Virginia, e.g., under-immunization, attention deficit hyperactivity disorder, asthma, and adverse perinatal health indicators. Epidemiologic research is being used to guide interventions planned and enacted by the community, with evidence of improved outcomes in certain indicators.CONCLUSIONS: A vitally important, emerging role for epidemiologists in the 21(st) century is to empower communities to identify and effectively address their own health needs. Epidemiologic data not only evaluates health status, but should be used to guide community action. PMID- 11018350 TI - Paper vs. electrons. Epidemiologic publishing in a changing world. AB - PURPOSE: To present the parallel histories of epidemiologic and electronic publishing and consider positive and negative factors that might affect their amalgam.METHODS: We performed a quantitative assessment of the arc of epidemiologic publication from 1966-1999, using major self-designated epidemiologic journals as a sample, and of scholarly electronic publication from 1991-1997, based on current literature review. We use an online, paperless journal as a case study, and review selected information-technology opinion in the area.RESULTS: By traditional standards, growth in epidemiologic publication has been considerable, with the addition of six new journals since 1966. In contrast, scholarly electronic publication for the period 1991-1997 grew from 27 to 2459 journals (not all exclusively online). Positive features of electronic publishing include flexibility, shortened time to publication, freedom from fixed publication date, diversity in presentation, and instant linkage to relevant material. A case study of a new online journal illustrates the substantive power of the medium. Negative factors include restriction (or unrestricted expansion) of the audience, the potential for hasty peer review, pitfalls in establishing credibility, an emphasis on style over content, technologic dependence, and additions to the information explosion. Relative cost and archiving are still debated. In assessing the pros and cons, it is important to distinguish electronic mechanisms that facilitate publication from electronic publishing, and to appreciate the difference between moving an existing journal to the electronic medium, and creating a new online journal.CONCLUSIONS: The movement from print to internet is probably inexorable, but a headlong rush may be ill-advised. Several models for dual publishing now exist, with the expectation that many, including the journals that serve epidemiology, will do so. The ultimate configuration is difficult to predict, but likely to be shaped in large measure by market forces, technologic advances and to a lesser extent, by habit and aesthetics. PMID- 11018351 TI - The role of epidemiology in the committee on the costs of medical care. AB - Among the most important 20th century documents of American public health is the 1932 report of the Committee on the Costs of Medical Care (CCMC). In May 1927, leaders of medicine, public health, and the social sciences, met at the annual meeting of the AMA, to consider that, despite advances in medical sciences, a vast amount of preventable disease and unnecessary death existed. A committee of 50 men and women, the CCMC, was formed to investigate. Of the six public health members, three were epidemiologists. A spectrum of governmental and private organizations supported the CCMC and eight foundations provided funding. A staff of 51 conducted 26 studies, many epidemiological, to support its five categorical recommendations. Briefly these were: 1) Medical services, preventive and therapeutic, should be provided by interdisciplinary groups; 2) Basic public health services should be available to all without cost; 3) The costs of medical care should be placed on a group basis (insurance, taxation, or a combination thereof); 4) Study, evaluation, and coordination of medical and public health services should be important functions for local and state governments; and 5) Professional education for a wide variety of medical and public health personnel should be greatly expanded with emphasis on prevention and social considerations.Perhaps the most interesting of the CCMC's epidemiological studies was, "The Incidence of Illness and the Receipt and Costs of Medical Care Among Representative Families: Experiences in Twelve Consecutive Months During 1928 1931." This study included 8,639 families comprising 38,668 persons and described the then current status of medical care and public health practice in the U.S.A.The final report of the CCMC contained minority reports ranging from claims that the recommendations were too radical to assertions that they didn't go far enough! Nevertheless, they have been the focus for medical and public health concern and controversy for almost three-quarters of a century. Their roots in epidemiology should not be forgotten! PMID- 11018352 TI - Prevalence of youth access to alcohol, guns, illegal drugs, or cigarettes in the home and association with health-risk behaviors. AB - PURPOSE: To describe the prevalence of access to alcohol, guns, drugs, or cigarettes in the home and its association with related health-risk behaviors among adolescents.METHODS: We analyzed cross-sectional data from the 1995 in-home survey of the National Longitudinal Study on Adolescent Health which used a nationally representative school-based sample (N = 6,504) of adolescents in grades 7-12. We used logistic regression analysis, adjusted for gender, race/ethnicity and age, to examine the associations between access to alcohol, guns, drugs, and cigarettes in the home and the practice of risk behaviors involving those variables.RESULTS: Overall, 1,817 (28%) adolescents reported having easy access to alcohol in the home, 1,616 (25%) had access to a gun, 189 (3%) had access to drugs, and 2,067 (32%) had access to cigarettes. Associations were found between easy home access to alcohol and drinking during the past 12 months (Adj. OR = 2.16, 95% CI = 1.89-2.47), ever being drunk at school (Adj. OR = 2.33, 95% CI = 1.85-2.95, and ever driving drunk (Adj. OR = 1.64, 95% CI = 1.29 2.09). Access to a gun at home was associated with carrying a gun to school (Adj. OR = 2.54, 95% CI = 1.40-4.64). Associations were also found between access to drugs and cigarettes in the home and ever using drugs and smoking regularly.CONCLUSIONS: Easy access to alcohol, guns, and cigarettes in the home is prevalent among adolescents and may increase involvement in risky behaviors. Limiting access therefore is important in order to reduce the occurrence of health-risk behaviors associated with substance use, deliquency and injury among adolescents. PMID- 11018353 TI - Frequency, characteristics, and child and parent risk factors for nonfatal injury in rural children. AB - PURPOSE: We sought to identify child and parent risk factors for nonfatal unintentional injury among rural children.METHODS: Design: cross-sectional health interview survey. Setting: a completely rural midwestern county not adjacent to a metropolitan area. Participants: stratified, random sample of farm, town, and nonfarm, nontown households, including parents of all children aged 0-17 years. Outcome measures: parent-reported injury episodes that occurred in the past 12 months in children that required professional care, restricted normal activities for at least four hours, or resulted in blacking out or losing awareness or memory. Potential risk factors included nine child factors and six parent factors.RESULTS: Of the 621 children in participating households, 137 or 22.1% were injured. Children on a sports team were 1.88 times (95% confidence interval: 1.07 to 3.31) more likely than other children to be injured. Track team members were more likely than non-members to have an injury (54.5% vs. 23.0%, chi square test, p = 0.001), and volleyball team members were more likely than non-members to have an injury (50.0% vs. 29.0%, chi square test, p = 0.009). Unrelated to injury status were the child's gender, rural stratum, emotional health, behavioral problems, impairment or medications. Also unrelated to child injury were the parent's marital status, alcohol dependence, binge drinking, antisocial personality, depression symptoms or fear of child abuse by spouse.CONCLUSIONS: Our findings illustrate the difficulty of discovering clues with potential to aid design of effective childhood unintentional injury prevention policies, programs, and practices. The instruments and processes in our study were of acceptable quality, therefore we failed to asses the true risk factors for childhood injury or our sample size was too small to detect associations with the factors that were included. PMID- 11018354 TI - Analysis of switching insurance plan type. Comparison of two statistical methods. AB - PURPOSE: To compare results of 2 statistical methods for identifying factors in claims data that are associated with switching insurance plans between managed care (MC) and indemnity (IN).METHODS: Using claims data from 2 insurance providers in a northeastern city, we analyzed patients aged 18+ with diabetes, asthma, or congestive heart failure (CHF) who were covered any time in 1993-1997 (N = 88,917). Stratifying by initial plan type, we examined predictors of switching from the initial plan type using logistic regression and survival analysis. Covariates included age, time in study (for logistic models), gender, diabetes (yes/no), CHF (yes/no), and asthma (yes/no). Survival analysis accounted for time to switch and allowed time-varying covariates.RESULTS: In logistic regression models, older individuals who were in IN were much less likely to switch into MC. Those in MC were more likely to switch to IN, with the greatest likelihood of switching in ages 60-69 (OR = 4.00, 95% CI = 3.32-4.83). Females were less likely to switch from IN to MC (OR = 0.92, 95% CI = 0.87-0.98), CHF patients were less likely to switch from IN to MC (OR = 0.75, 95% CI = 0.68 0.83), and diabetes patients were less likely to switch from MC to IN (OR = 0.77, 95% CI = 0.62-0.96). Hazard ratios calculated using Cox regression were similar to odds ratios for most covariates. However, some coefficients for diseases were significant in Cox models but not in the logistic models. Cox models took 45 times longer in CPU time than logistic regression models.CONCLUSIONS: Logistic regression was a good approximation to Cox regression in identifying many of the factors in switching insurance plan in these data, at a fraction of the computing time. However, Cox models allowed diseases to be time-varying, and so was more sensitive to identifying significant relationships with disease. PMID- 11018355 TI - Identifying persons with chronic conditions in a managed care setting. A comparison of methods. AB - PURPOSE: Together, the sophisticated computerized information systems and defined, stable populations within Health Maintenance Organizations (HMOs) can reduce imprecision associated with modeling disease incidence and prevalence. Such data are useful for resource allocation, intervention planning, epidemiological study and surveillance. Identifying all cases of illness within such a defined population is vital to these aims, yet detailed diagnostic information is lacking in many pre-paid plans. In this presentation, diagnostic and prescription data are compared for their ability to assess the treatment prevalence for 28 chronic medical conditions.METHODS: Group Health Cooperative is a mixed model HMO in Western WA state. Automated records for calendar year 1999 were examined for 182,174 commercially insured patients age 20-64. Prescription drug data and International Classification of Disease-9(th) Clinical Modification codes (ICD-9CM) were used to identify 28 chronic medical conditions among patients. Chi square tests were performed to compare results produced by the two methods.RESULTS: The diagnostic method worked best for conditions with no clear pharmacologic treatment. However, the pharmacy-based method better identified patients with chronic illnesses initially diagnosed prior to 1999, and with disorders for which standard diagnostic codes are inconsistently utilized, such as mental health conditions.CONCLUSIONS: While both diagnostic and pharmacy-based methods perform well in identifying a limited set of conditions, each performs poorly under certain circumstances. A hybrid method utilizing both diagnostic and pharmacy data may be more appropriate for an HMO setting. Additional research is needed to develop and refine such a method. PMID- 11018356 TI - Insights from a large rural population laboratory. Health census '89 and '99. AB - PURPOSE: Data are limited on health status behaviors and use of health services for rural residents. Yet rural areas now have higher rates of chronic diseases, such as coronary heart disease, than urban areas.METHODS: A population laboratory (Health Census) was established in rural Central NY (Otsego County) to study these variables in this population. HC '89 was a door-to-door enumeration of all permanent households in the country. In 1989, 44,500 adults in 18,000 households provided data on individual health screenings, cancer and cardiovascular risk factors, health behaviors, chronic disease, access to care, preventive services use, health insurance and emergency department utilization. Data collection for HC '99 will identify 10-year trends; it also includes baseline data for children. Also, households in six surrounding rural counties (N = 10,000) were sampled; cost benefits of different survey methods were assessed. Questions about perceived health status, and special health and pediatric preventive care needs were included. After 10 years of social change, many limitations of traditional survey methods became apparent; much more varied and frequent types of follow-up were needed to achieve adequate response rates.RESULTS: The HC '89 final response rate was 86%. There were significant socioeconomic gradients in use of preventive services (blood pressure screening, rectal exams, mammograms and Pap smears). Adults without health insurance or Medicaid had much lower utilization rates of screening tests and higher rates of cigarette smoking and obesity. Several community intervention programs were implemented as a result of HC '89. HC '99 provides additional analyses to examine change in preventive service use, self reported chronic disease, and health services use; it will likely verify the persistence of socioeconomic gradients. Response rates are similar to '89, but achieved only with more intensive reinforcement.CONCLUSIONS: HC '89 confirmed a lag in positive health indices in rural populations in Central NY. Both newer methodologies now needed and results of HC '99 will serve as guides for smaller rural counties to develop affordable local health surveys and plan intervention strategies. PMID- 11018357 TI - Characteristics of chronic arthritis and other rheumatic condition-related ambulatory care visits, united states, 1997. AB - PURPOSE: To characterize ambulatory medical care visits among persons with arthritis and other rheumatic conditions, the leading cause of disability.METHODS: The 1997 National Ambulatory Medical Care Survey (NAMCS) and National Hospital Ambulatory Medical Care Survey (NHAMCS) collect annual data on the utilization of ambulatory medical services provided by non-federal office based physicians and hospital outpatient and emergency departments. Arthritis related visits were defined using a predetermined set of ICD9-CM diagnostic codes developed by an expert panel and designed to include all potential diagnoses for arthritis and other rheumatic conditions. Visits related to acute conditions such as injuries were not included. National estimates and rates of arthritis-related ambulatory care visits were calculated by age, race, and sex groups.RESULTS: In 1997, there were an estimated 959.3 million ambulatory care visits, of which over 38 million (4.0%) were related to arthritis and other rheumatic conditions. Arthritis-related visits were more likely to be made by females (65.4%), white persons (82.2%), non-Hispanic persons (72.7%) and persons aged 25-64 years (61.9%). More than one-third of arthritis-related visits were for osteoarthritis, rheumatoid arthritis and unspecified myalgia/myositis. About half (50.2%) of the office visits for arthritis were made to general/family physicians or internists, while an additional 16.2% were to rheumatologists. Counseling or education related to exercise, diet/nutrition and injury prevention were provided at 18.9%, 9.2% and 2.2% of office and outpatient department visits respectively.CONCLUSIONS: Arthritis and other rheumatic conditions are common conditions associated with ambulatory medical care. These results suggest missed opportunities for counseling patients regarding public health prevention messages for arthritis, including increasing moderate physical activity, weight management and injury prevention. PMID- 11018358 TI - Comparison of utilization of preventive health care services between two racial populations. AB - PURPOSE: The difference in utilization of existing preventive health care services (PHCS) may be related to race. We compared the difference in use of PHCS between black and white males in Northwest Louisiana.METHODS: A cross-sectional survey regarding utilization of preventive health care services was conducted from 09/93-10/97. Approximate equal number of white and black males participated in this quota survey.RESULTS: 70% of black males said they had ever gone to the doctor for routine preventive check-up, while 79% of white males did so (p < 0.001). For those who previously had a routine preventive check-up, 88% had their last one within one year. Most routine check-ups are provided as part of the employment. The white males are 1.4 times more likely to have a routine check-up as compared to black males (p < 0.05). Factors which have a positive effect on use of PHCS are education (OR = 1.07, p < 0.01); intent to get a check-up (OR = 9.0, p < 0.001); the doctor spending enough time with the patients (OR = 2.4, p = 0.01); and wanting some health information (OR = 1.7, p < 0.001). Factors which have a negative effect on use of PHCS are cost (OR = 0.73, p = 0.046); having to go to different places (OR = 0.65, p = 0.024); and being scared of the doctor (OR = 0.46, p < 0.001). An indication of a negative effect-although not statistically significant-was found for long waiting time (OR= 0.73, p = 0.55) and the doctor not spending enough time explaining to patients (OR = 0.47, p = 0.054).CONCLUSIONS: White males that are more likely to use PHCS as compared to African-Americans. People of higher education with a positive intent and actively seeking health information are more likely to use PHCS. The more time the doctor spends with the participant, the more likely it is the participant will come for a routine check-up. The barriers for use of PHCS are cost, having to go to different places, and being scared of the doctor. PMID- 11018359 TI - Smokers eligible for a clinical trial. Correlates of not returning for randomization. AB - PURPOSE: Recruitment and retention of minority participants is often the rate limiting step towards successful completion of clinical trials. Participants who are eligible, and express interest in, enrolling into a trial will sometimes not return for their randomization visit. Factors associated with failure to return for randomization are largely unknown.METHODS: We compared 287 eligible African American (AA) smokers who enrolled, but did not return for randomization (NR), to 500 AA smokers who returned and were randomized (R) to participate in a study comparing culturally sensitive educational materials to usual care materials for smoking cessation in AAs. An analysis was conducted on variables potentially associated with not returning for randomization.RESULTS:Table 1CONCLUSIONS: Potential NR participants differed in a number of ways from those who returned for randomization. Better understanding of these factors may allow researchers to target recruitment efforts, potentially resulting in enhanced accrual and retention, and therefore, generalizability. PMID- 11018360 TI - Selection of controls in injury case-control studies. AB - PURPOSE: To assist in elucidating principles underlying the design of injury case control studies.APPROACH: We begin by defining "event" as the sequence of circumstances that place a person at risk of injury (e.g. bicycle crash) and "injury given the event" as the resultant physical damage (e.g. head injury in bicycle crash). We then identify two broad classes of research question: 1) Studies of risk factors for the event, and, 2) Studies of risk factors for injury given the event. The study base for the first type of research question is all persons at risk of the event, while the study base for the second type is all persons who experience the event, and are therefore at risk of injury. It follows that in studies of risk factors for injury given the event, the controls should be a sample of all persons who experience the event. For example, in a study of bicycle helmets and head injuries, a suitable case group would be cyclists treated for head injury following a bicycle crash. The appropriate control group is drawn from the base population of all cyclists who crashed, including those who had no injuries. The control group may be restricted to cyclists who crashed and sought treatment for non-head injury under the assumption that the exposure distribution (prevalence of cycle helmet use) in the crash/no injury group is identical to the exposure distribution in the crash/non-head injury group.CONCLUSIONS: It is over ten years since innovative researchers in Seattle first applied the case-control design to the problem of bicycle crashes. Since then, successive bicycle injury studies at other centers have largely failed to extend and refine the Seattle methodology. A more critical approach to the design of case-control studies is required if we are to continue to advance the field of injury epidemiology. PMID- 11018361 TI - A common measure of disease-gene associations for case-control and tdt studies. AB - PURPOSE: Associations between a disease and a candidate gene can be evaluated in case-control studies, or using the transmission/disequilibrium test (TDT) in data from studies of case-parent trios. Consequently, data for a given disease-gene association are often available from both case-control and "TDT studies." The present investigation was undertaken to identify a measure of the magnitude of a disease-gene association that could be estimated using data from either type of study.METHODS: The magnitude of a disease-gene association detected using the TDT can be estimated by the frequency (T) with which heterozygous parents transmit the putative, high-risk allele to affected offspring. To derive an expression for T that could be solved using data from a case-control study, T was first rewritten as a function of the relative frequencies of the genotypes, at the candidate gene, among the affected offspring of a heterozygous parent. By application of Bayes' theorem, this equation was reduced to a form that could be solved using case-control data.RESULTS: T can be expressed as a function of the genotypic probabilities among individuals with the disease of interest and the frequency of the high-risk allele in the general population, which can be estimated from cases and controls, respectively. Hence, estimates of T can be derived from the data available in a case-control study.CONCLUSIONS: The parameter T, which provides a measure of the magnitude of a disease-gene association, can be estimated from either case-control or TDT data and, therefore, provides a means by which to compare results from these two types of studies. As in other areas of epidemiology, comparison of a quantitative measure of the magnitude of an association, as well as the direction and statistical significance of the association, should greatly aid in the interpretation of the overall body of literature on a particular disease-gene association, and may help to clarify seemingly discrepant results. PMID- 11018362 TI - Willingness to participate in biomedical research. African-americans vs whites. AB - PURPOSE: The overall goals of this study were: 1) to determine whether (and if so, the extent to which) African-Americans differed in their willingness to participate in biomedical studies as compared to non-Hispanic Whites, and 2) to determine the influence that a range of demographic, psycho-social and medico historical variables had on the comparative willingness of these groups to participate in biomedical research. This report will present preliminary findings on the first goal.METHODS: The Tuskegee Legacy Project (TLP) Questionnaire, a 60 item instrument developed within the Northeastern Minority Oral Health Research Center (a P50 Center supported by the National Institute of Dental and Craniofacial Research at NIH), was administered via a random-digit dial telephone interview by the University of Alabama at Birmingham Survey Research Unit to 840 adult African-Americans and Whites in three city/county areas: Birmingham/Jefferson Cty, AL; Hartford/Hartford Cty, CT and Tuskegee/Macon Cty, AL.RESULTS: The response rates were 70%, 65% and 49% for Birmingham, Tuskegee and Hartford, respectively. A lower percentage of African-Americans (21%) reported that they were either somewhat or very likely to participate as biomedical research subjects than did Whites (29%) (p < 0.0001). In response to 7 prompts on 'who' was conducting a study, African-Americans indicated they were less likely to participate than Whites on 4 specific prompts and more likely to participate on only 1 prompt (p < 0.006 for each). In response to 9 prompts on 'what' a study asked participants to do, African-Americans indicated they were less likely to participate than Whites on only 2 specific prompts (p < 0.001 for each), more likely to participate on 2 prompts (p < 0.001), and were equally or near equally likely to participate on 5 prompts.CONCLUSIONS: The findings from this study show that African-Americans self-reported a lower willingness to participate in biomedical studies than did non-Hispanic Whites, and that African-American subjects were more influenced by the factor of 'who' was running the study than by the factor of 'what' they might have to do as study subjects. Study supported by grant P 50 DE10592 from the NIDCR at NIH. PMID- 11018363 TI - Physical activity surveillance in the 21(st) century. AB - PURPOSE: The 1996 Surgeon General's Report on Physical Activity and Health emphasized the health-related benefits of moderate-intensity physical activities, especially everyday activities. Unfortunately most surveillance systems in the U.S. primarily measure sports-related activities, particularly vigorous intensity sports. This report describes a new physical activity surveillance instrument designed to go beyond our current measures and include moderate-intensity everyday activities.METHODS: Data were collected from a nationally representative sample of adults (n = 5010). Questions were asked about occupational activity (mostly sitting; mostly walking; mostly heavy labor); walking (for exercise, transportation, or any other reason); moderate-intensity activities (brisk walking, yard work, vacuuming); vigorous-intensity activities (running, aerobics, heavy yard work); and strengthening activities (lifting weights, pull-ups, sit ups). Questions included frequency and duration of activities. Respondents were defined as recommended if they participated in either moderate-intensity activities >/=30 min/day for >/=5 days/wk OR vigorous-intensity activities >/=20 min/day for >/=3 days/wk. Insufficient was defined as not meeting recommended levels while inactive was defined as no leisure-time activity.RESULTS: Overall 40% of adults were in the recommended group, 44% were in the insufficient group and 16% were inactive. Among working adults 37% had jobs that involved mostly walking or heavy labor and about 50% of those also were in the recommended group. Among the 63% of working adults who report mostly sitting at work, 39% were in the recommended group.CONCLUSIONS: These results suggest that measuring only leisure-time physical activity may under represent the physical activity experience of many U.S. adults. Broadening the concept of physical activity beyond traditional sports-related vigorous "exercise" may provide a more accurate picture of the prevalence of health-related physical activity. PMID- 11018364 TI - Digit preference in year at menopause. Data from the study of women's health across the nation. AB - PURPOSE: To assess possible digit preference in self-reported year at natural menopause, and to determine whether equal proportions is an appropriate reference distribution.METHODS: Data are from the cross-sectional telephone interview from the Study of Women's Health Across the Nation (SWAN), a multi-site, multi-ethnic study of women aged 40-55. Analyses included 2151 naturally menopausal respondents, who were asked the year their periods stopped. Using a chi-square test, we compared the distribution of the terminal digit for year of final menstrual period (FMP) to an equal proportions (EP) distribution assigning 10% probability of each of the 10 digits. Departures from EP, however, may be accurate and may reflect the observed age distribution of subjects rather than digit preference. Thus, we stratified by year of interview to determine if the distribution was the same across years-suggesting digit preference-or if it varied systematically. We then estimated an expected distribution of terminal digit for year of FMP, using prospectively collected data (not subject to digit preference or recall bias) from the Massachusetts Women's Health Study, applied to the SWAN age distribution. The SWAN terminal digit distribution was compared with this expected distribution.RESULTS: Terminal digit for year at FMP exhibited a strong departure from EP (chi(2) = 841.60, p < 0.001), with highest frequencies for digits 4 and 5. Stratifying by year of interview, the distribution was shifted one digit in 1997 compared with 1996, consistent with unbiased reporting. Using the expected distribution as the reference reduced the chi-square statistic by a factor of 7 (chi(2) = 119.51, p < 0.001).CONCLUSIONS: The distribution of terminal digits for reported year at FMP was far from uniform, but departures from EP were consistent with results expected from an independent prospective study. These results suggest that EP may not be an appropriate reference for studies of digit preference, particularly those with a restricted age range. PMID- 11018365 TI - Trends in reproductive, smoking, and other chronic disease risk factors by birth cohort in a large occupational study. AB - PURPOSE: To illustrate the value of using large cohort studies to identify birth cohort trends in several chronic disease risk factors.METHODS: In collaboration with the American Registry of Radiologic Technologists (ARRT) and the University of Minnesota, the National Cancer Institute (NCI) initiated a cohort study of radiologic technologists who were certified by ARRT for at least two years between 1926 and 1982. Over 90,000 technologists (nearly four-fifths female) from all 50 states responded to a mailed questionnaire on reproductive, medical, work, and lifestyle factors. Ten, mostly five-year, birth cohorts, from before 1920 through 1960 and later, were evaluated.RESULTS: In this population, the mean height of both men and women generally rose in each subsequent birth cohort. The proportion of men who smoked before age 18 fell among those born since the late 1920s. In contrast, the proportion of women smoking before age 18 rose among those born since the early 1950s, reaching 14.2% among those born in 1960 and later. The mean age at menarche fell, until leveling off at 12.5, among those born after 1940. Recent birth cohorts (since 1950) show among the highest mean ages at birth of first child (>26 yeras), highest rates of nulliparity at age 25 (>/=63%), and lowest mean parity levels (/=6 months and severe symptoms. PMID- 11018369 TI - Are trends in diabetes incidence changing for minority children? AB - PURPOSE: To contrast incidence rates and time trends of typical type 1 diabetes with non-autoimmune insulin-resistant type 2 diabetes in minority children (Early 2).METHODS: A population based registry of all insulin treated African-American and Hispanic children residing in Chicago, aged 0-17 years at onset, diagnosed between 1985-1994 provided data. Medical records were reviewed and abstracted (N = 739), and a subset of patients were interviewed (n = 195). Based on these data sources, cases were assigned to the early type 2 category if one or more of the following variables were present: notation of "atypical" or "possible type 2" diabetes in the medical record; obesity (defined as BMI >/= 27 (kg/m(2))) or acanthosis nigricans noted at diagnosis; a report by the patient of going off insulin six months after initial diagnosis without developing diabetic ketoacidosis; or reporting the use of oral hypoglycemic agents with or without insulin. The Chi-square test for homogeneity was used to determine differences between proportions of groups. Time trend analysis of incidence was done using regression models with the Poisson distribution.RESULTS: The 10-year average annual incidence rate was 10.3 per 100,000 population for typical type 1 diabetes (n = 565) and 3.2 per 100,000 for those with characteristics of early 2 diabetes (n = 174). Over the ten year interval the incidence rate remained static for typical type 1 diabetes. In the early 2 group there was a 9.0% average annual percent increase over the ten year interval (p < 0.01).CONCLUSIONS: The trends in incidence rates vary between typical type and early 2 diabetes. We hypothesize that this is associated with the increasing rate of obesity and a lack of physical activity in young people. PMID- 11018370 TI - The impact of birth weight on the racial disparity of end-stage renal disease. AB - PURPOSE: Blacks have a high rate of end-stage renal disease (ESRD) and low birthweight (LBW) than whites. LBW has been associated with ESRD. The purpose of this study was to assess impact of LBW on the racial difference in ESRD.METHODS: Patients born in SC after 1950 and diagnosed with ESRD between 1991-1996 were identified from the ESRD registry. Birth weight was compared for 858 black and 372 white patients and 2460 controls matched for age, sex, and race. LBW was defined as birthweight <2500 g.RESULTS: Among patients with ESRD, mean birthweight was lower in blacks than whites (3179 vs 3367 g, p < 0.001). LBW was more common in blacks than whites with ESRD (13.9 vs 7.5%, p = 0.02). The risk ratio for LBW among ESRD patients was 1.4 (95% C.I. 1.1 to 1.8) for blacks and 1.5 (95% C.I. 0.9 to 2.5) for whites. The population attributable risk (PAR) for ESRD due to LBW was greater for blacks than whites (33.6 vs 4.2 per 100,000).CONCLUSIONS: Birthweights were lower and LBW was more common among blacks than whites with ESRD. Moreover, LBW contributed more to the PAR of ESRD in blacks than whites. Thus, LBW may contribute to the greater risk for ESRD in African Americans than Caucasians. This preliminary study indicates that further research on the link between LBW and ESRD could be instructive in understanding the racial health disparities. PMID- 11018371 TI - Trends in the rate of emergency room admissions for preventable cardiovascular conditions among african american men and women over the past decade. Continuation of negative trends. AB - PURPOSE: The objective of this retrospective analysis was to compare secular changes in the rate of emergency room admissions (per 100,000) for selected acknowledged preventable cardiovascular conditions among African Americans (AA) men and women aged >/=21 from 1991-1998, and rate of change for Caucasian (Cau), Hispanic (Hisp), and Asian (Asi) men and women aged >/=21; conditions included angina, congestive heart failure (CHF), diabetes, and hypertension.METHODS: Results are derived from calendar-year California hospital data based on a selection of specified ICD-9 codes that correspond to the principal diagnosis for admission. The combined study sample size included a total of 21,016 individuals who were admitted to a hospital via the ER. Separate standardized and age adjusted Poisson regression models were employed for each condition to assess race and time main effects and race x time interaction terms (P 100,000). Statistical analyses were performed with SAS Version 8.RESULTS:Table 1The rates of SR, TB, and BR were significantly higher in the coastal region. In contrast, no coastal pattern was detected by population density.CONCLUSIONS: The consistent association of SR, TB, and BR with geographical region, as opposed to the inconsistent association with population density, suggests that environmental factors may be involved in these pulmonary diseases. These results demonstrate the need for further analysis of the relationship between socio-economic, behavioral, and environmental factors and the development of these diseases. PMID- 11018374 TI - An epidemiological case-control study of migraine and its associated comorbid conditions. AB - BACKGROUND: The economic impact of migraine to society has been well documented. However, few studies have identified the risk of migraine-specific comorbid conditions to explain the higher healthcare resource utilization by migraineurs. Because comorbidity has the potential to increase treatment costs, estimating its risks and its economc impact becomes imperative.OBJECTIVES: To estimate the risk for migraine by presence of a defined set of comorbidities, and to quantify the association of estimated healthcare costs between the case-control groups with and without comorbidity.METHODS: A matched case-control was designed using data from 1998 Idaho Medicaid claims. Cases were identified by physician-diagnoses and migraine-specific medication use. A 1:1 matching was performed on age, race, gender, and residence. Presence of comorbidity was determined by ICD-9 diagnostic categories. Total healthcare resource utilization was estimaed for all physician, hospital (inpatient and outpatient), and emergency services, including prescription medicines paid by Idaho Medicaid. Multivariate logistic regression was used to compute the odds ratio controlling for the matching variables. Multiple-regression models were used to quantify the association of total healthcare costs.RESULTS: Migraine patients were approximately five times more likely (Odds Ratio = 5.34; Confidence Interval(95%)-4.38 to 6.52) than their controls to be diagnosed with comorbid condition(s). A model with variables of age, gender, presence of comorbidity and migraine, explained approximately 21% of the variance in healthcare utilization (p < 0.05).CONCLUSIONS: Migraine patients are at a higher risk for defined comorbid conditions resulting in significantly higher healthcare costs as compared to non-migraine patients. PMID- 11018375 TI - Prior hrt use, ethnicity and mammographic density. AB - PURPOSE: Mammographic density has been strongly associated with breast cancer risk in several studies. To clarify possible pathogenic mechanisms underlying this relationship, we assessed factors affecting mammographic density in postmenopausal women.METHODS: Percent density (PD) was measured on pre randomization mammograms from 425 participants in the Women's Health Initiative clinical trial; no women were currently using HRT. Trained observers used a previously validated computer-assisted thresholding technique to measure PD (ratio of dense areas to breast area) on craniocaudal films. Univariate relationships were assessed between PD and age, parity, education, history of HRT use, age at first birth, abortion history, alcohol use, serum cholesterol, physical activity, smoking, dietary fat intake, body-mass index (BMI), and waist to-hip ratio (WHR). Multivariable regression was used to identify predictors of PD (log transformed), controlling for confounders.RESULTS: PD ranged from 0-60% with mean 8%. Age, gravidity, BMI, and WHR were negatively associated with PD. Mean PD in the highest tertiles of BMI and WHR was nearly half that in the lowest. Ethnicity (50% white, 28% black, and 19% Hispanic) and hysterectomy status (50% had intact uterus) were unassociated with PD. Controlling for age, BMI, WHR, smoking, hysterectomy status, prior HRT, and nulliparity, mean PD differed significantly by ethnicity, with black women having highest PD. However, the effect of prior HRT use differed by uterine status. In women with uteri intact, ethnicity, but not prior HRT use, was related to PD after controlling for age, BMI, and WHR. In hysterectomized women, ethnicity was unrelated to PD, while previous HRT was associated with lower PD.CONCLUSIONS: Gravidity, ethnicity and body shape and size are associated with mammographic density in postmenopausal women. Previous hormone use predicts lower PD, but only among hysterectomized women. This may be due to the higher frequency of oophorectomies in this group, which may lower PD and increase likelihood of HRT use. Planned analyses of entire sample of 1050 women will clarify the effect of these and other factors on density, and on density change in followup mammograms. PMID- 11018376 TI - Ethics and breast cancer risk assessment. AB - Clinical risk assessment holds great promise for identifying individuals who might benefit from preventive interventions. In the case of breast cancer, statistical models, notably the Gail model, have been developed to assess an individual woman's future risk of developing disease. However, the estimates derived from these models are subject to substantial uncertainty and there is controversy over how to translate risk information into prevention and control measures.In light of these uncertainties, ethical concerns have been raised about appropriate use of these models. The potential benefits of individualized risk assessment must be weighed against current limitations and potential harms. The fact that breast cancer is a significant source of anxiety for many women suggests that the potential harms from misinformation are substantial.This paper concludes that the profound uncertainties surrounding breast cancer risk assessment warrant caution in the use of such models. Breast cancer risk assessment tools occupy a grey area between public health education and individualized clinical attention. When public health officials promise individualized risk information, there is potential for women to place too much importance and trust in these risk estimates. Moreover, use of these models in counseling women about participation in clinical trials should be responsive to the complexity of informed consent. PMID- 11018377 TI - The relation between hip fracture and alzheimer's disease in the canadian national population health survey health institutions data, 1994-1995. A cross sectional study. AB - PURPOSE: The purpose of this study is to examine the relation between hip fractures and Alzheimer's disease in institutionalized men and women who participated in the 1994-1995 Canadian National Population Health Survey (NPHS).METHODS: Participants in the institutional component of NPHS were randomly chosen from selected health care institutions from all provinces in Canada. A questionnaire, which assessed health, demographic and socio-economic status, risk factors, medication use, and falls, was administered by an interviewer. Proxy respondents were sought for residents who were ill or incapacitated. Logistic regression was used to examine the association between hip fractures and Alzheimer's disease in 408 men and 1105 women >/=65 years. Models were examined with either hip fracture or Alzheimer's disease as the dependent variable. Covariates that were assessed included osteoporosis, age group, sex, medications, reported falls and comorbid conditions.RESULTS: All hip fractures reported in this survey were the result of a fall, however only 3.7% of falls resulted in a hip fracture. Those who had sustained a hip fracture were more likely to have Alzheimer's disease (OR 2.0, 95% CI 1.1-3.5), osteoporosis (OR 4.3, 95% CI 2.5 7.4) and heart disease (OR 2.4, 95% CI 1.1-5.0). Respondents who had Alzheimer's disease were more likely to have sustained a hip fracture (OR 2.1 95% CI 1.2 3.6), to have osteoporosis (OR 1.9, 95% CI 1.5-2.5), and to have fallen (OR 1.4, 95% CI 1.1-1.8) and were less likely to be taking anti-psychotic medication (OR 0.4, 95% CI 0.3-0.6) than those with no diagnosis of Alzheimer's disease.CONCLUSIONS: There is an association between Alzheimer's disease and hip fractures that is independent of other covariates in this representative sample of institutionalized elderly Canadians. PMID- 11018378 TI - The metropolitan new york registry & cfrbcs. Unique resources for breast cancer research. AB - PURPOSE: The goal of the Metropolitan NY Registry, and five other international collaborating sites of the Cooperative Family Registry for Breast Cancer Studies [CFRBCS], has been the development of a comprehensive resource for interdisciplinary genetic epidemiology studies addressing breast cancer risk and prognostic factors.METHODS: Family recruitment has been conducted in clinical and community settings by a multidisciplinary team of collaborators affiliated at six major metropolitan NY medical centers. Families meeting one of the following criteria were invited to join: a male with breast cancer; a female with breast or ovarian cancer diagnosed at age 3,000 epidemiology questionnaires completed. Ethnic/racial data indicates 14% are of African American or Hispanic heritage and 509 families (46%) are of Ashkenazi descent. One or more DNA samples from 400 Ashkenazi families have been tested for the three BRCA1/2 founder mutations. Of 331 Ashkenazi participants with a history of breast and/or ovarian cancer, 19% were found to be mutation carriers. Thirty-six (7 men/29 women) mutation carriers are free of cancer.CONCLUSIONS: The families participating in the NY Registry reflect a spectrum of breast cancer risk. The extensive NY and CFRBCS databases and banked biospecimens provide a unique resource for multidisciplinary genetic epidemiologic studies that may identify avenues for prevention. PMID- 11018379 TI - An intervention study on screening for breast cancer among single african american women aged 65 and older. AB - PURPOSE: Older single African-American women are the population that is least likely to use screening procedures because of cognition-related, income-related, social-support-related and medical care-related barriers. This study aims to evaluate a breast screening intervention program developed according to socioeconomic, cultural, psychological and behavioral characteristics of older single African-American women.METHODS: Ten public housing complexes were randomly assigned to either intervention or control group. African-American women aged 65 and over were recruited into the study if they were widowed, divorced, separated or never-married in the preceding year, and did not have a history of breast cancer (n = 325). Delivered by lay health educators, the intervention program targeted increasing knowledge on breast health and breast screening, reducing emotional or psychological problems, and increasing support from the significant others of study women. Breast screening-related cognition and behavior were measured at pre-intervention and post-intervention.RESULTS: Comparisons of the pre-intervention and post-intervention measurements showed that while the proportion of women who had a clinical breast examination or mammogram in the preceding year was decreased at the post-intervention in the control group, it was increased in the intervention group. However, the differences did not reach a significant level. No consistent patterns could be found in changes of variables in knowledge, attitudes and beliefs. These results remained similar when potential confounding factors were adjusted using mixed model regression analyses.CONCLUSIONS: These results did not suggest significant effects of an intervention program which used lay health educators to promote breast cancer screening in older single African-American women. PMID- 11018380 TI - Factors associated with accurate cancer detection during a clinical breast examination. AB - PURPOSE: Although most US women who undergo mammography screening also receive a Clinical Breast Examination (CBE), factors that predict cancer detection during a CBE have not been well studied to date.METHODS: We analyzed 752,081 National Breast and Cervical Cancer Early Detection Program (NBCCEDP) screening records for CBEs performed from July 1, 1995 thru June 30, 1998 on low income women across the US. CBE results are reported as "normal" or "abnormal, suspicious for cancer." Other variables include CBE date, age, race, ethnicity, presence of breast symptoms at screening, and whether the woman had received a prior NBCCEDP funded CBE. Most (74%) records include mammography dates and results. Tumor stage and size are reported for invasive cancers whether detected by CBE or mammography.RESULTS: Breast cancer was reported on 3,780 records. On 2,224 records, the CBE was suspicious for cancer. On 1,556 records, the CBE was normal. Most (93%) cancer records with a normal CBE had abnormal mammography results. Bivariate comparisons between normal and abnormal CBE records with a cancer diagnosis found significant differences (p <.001) in age, race/ethnicity, presence of breast symptoms, history of a prior NBCCEDP CBE, and tumor stage and size. In a logistic model controlling for these variables, women with breast symptoms were more likely to have their cancer detected during CBE (OR = 7.0, 95% CI = 5.9-8.5), while women with a prior NBCCEDP CBE were less likely to have their cancer detected (OR = 0.7, 95% CI = 0.6-0.8). Compared to women aged 50-59 years, women <40 were more likely to have their cancer detected by CBE (OR = 3.2, 95% CI = 1.9-5.4) but women >/=70 were less likely (OR = 0.7, 95% CI = 0.5-0.9). Larger cancers and those at more advanced stages were much more likely to be detected during CBE.CONCLUSIONS: Among low-income women whose cancers were detected through the NBCCEDP, older women and those who had been screened previously in the program were less likely to have their cancer detected during a CBE, even after controlling for tumor size and stage. PMID- 11018381 TI - Depression-related variables and breast self-examination in single older african american women. AB - PURPOSE: Depression-related symptoms, such as despair and loneliness, may prevent women from using preventive screening for diseases. This study aimed to assess if depression-related variables affect the likelihood of implementation of breast self-examination (BSE) in single African-American women aged 65 and older.METHODS: Subjects for this study included 325 African-American women, aged 65 and older, who were widowed, divorced, separated, or never-married, and lived in ten public housing tenements in Nashville, Tennessee. In-person interviews were conducted to collect information on breast screening behavior, knowledge and attitudes, social networks and activities, medical care use and depression. Depression variables included 19 factors, such as feeling guilty, feeling sad or blue, and feeling worthless.RESULTS: Using logistic regression with adjustment for potential confounders, we found that, compared with women who did not perform BSE, those who performed BSE were less likely to have felt guilty in the preceding year (p < 0.05, odds ratio (OR) = 0.3, confidence interval (CI) = 0.13 0.71). However, the other depression-related variables were not associated with BSE. For example, women who performed BSE and those who did not perform BSE reported feeling sad or blue in equal frequencies (OR = 0.93, CI 0.49 1.7).CONCLUSIONS: In general, this study suggests that there is no overall association between depression-related symptoms and breast self-examination in single African-American women aged 65 and older. PMID- 11018382 TI - Racial differences in tamoxifen metabolism. AB - PURPOSE: A recent study indicates that black women may have a higher risk/benefit ratio from tamoxifen than white women. Several reports in the literature indicate that racial differences in the risks/benefits from other pharmaceutical agents may be partially due to racial differences in drug metabolism. Thus, we explore the hypothesis that the racial difference in the tamoxifen risk/benefit ratio may be due, in part, to racial differences in tamoxifen metabolism.METHODS: We conducted a pilot study in which we recruited 6 white and 4 black breast cancer patients from the Baltimore, Maryland area. All patients were taking tamoxifen for at least 30 days. Each provided a blood sample that was used to measure tamoxifen metabolites by high performance liquid chromatography.RESULTS: Our results indicate that the black women had significantly higher levels of the tamoxifen metabolite, N-desmethyltamoxifen (N-DMT) than the white women (0.585 ug/ml vs 0.199 ug/ml, p < 0.05). There were no differences in the serum levels of tamoxifen in black and white women (0.809 vs 0.699, p > 0.1).CONCLUSIONS: These data suggest that both black and white breast cancer patients reach steady state tamoxifen levels, but that black women are more likely to metabolize tamoxifen to N-DMT or to maintain higher levels of N-DMT (i.e., less excretion of N-DMT) than white women. N-DMT is thought to be less effective in breast cancer treatment than tamoxifen. N-DMT also has been associated with excess proliferation in breast cancer cells. Thus, it is possible that the relatively high tamoxifen risk/benefit in black women occurs because tamoxifen is readily converted to N DMT in black women, but not in white women. Although the data reported here are from a pilot study, it is important to note that significant racial differences in tamoxifen metabolism were observed with only a few participants. We urge other investigators to confirm these findings using a large population of breast cancer patients. PMID- 11018383 TI - Mean arterial pressure, pregnancy induced hypertension, and preeclampsia. Evaluation as independent risk factors and as surrogates for high maternal serum alpha-protein in estimating breast cancer risk. AB - PURPOSE: Data from a nested case-control study were analyzed to examine high mean arterial pressure (MAP), hypertension of pregnancy and preeclampsia as independent predictors, and as surrogate markers for elevated Alpha-fetoprotein (AFP) levels in evaluating breast cancer risk.METHODS: Cases (n = 205) were identified by the California Cancer Registry from a cohort of pregnant women who were part of the Kaiser Health Plan and took part in the Child Health and Development Studies initiated by the University of California, Berkeley, from June 1959 to September 1966. Controls (n = 337) were selected by randomized recruitment from the same cohort probability matched to cases by distribution of birth dates of cases.RESULTS: High MAP was associated with breast cancer risk, and was different across quartiles of age at first full term pregnancy. Odds Ratios across quartiles were 0.24 (95% CI = 0.08-0.71), 0.84 (95% CI = 0.39 1.66), 1.00 (referent), and 2.50 (95% CI = 1.21-5.13). Neither diagnosed preeclampsia nor hypertension of pregnancy showed any association with breast cancer risk. When both high Alpha-fetoprotein and high mean arterial pressure were entered into the same analysis neither changed the odds ratio for the other more than eight percent. Additionally, AFP level was not a linear function of MAP.CONCLUSIONS: Although the pattern of odds ratios across quartiles of age at first full term pregnancy were similar for the two variables it can not be concluded that high MAP is an adequate surrogate for high levels of maternal serum AFP, but rather represents some related process which is in and of itself a risk factor for breast cancer. PMID- 11018384 TI - Cross-sectionally derived hysterectomy prevalence for correcting uterine and ovarian cancer incidence rates and probabilities. AB - PURPOSE: This paper presents a cross-sectional method for deriving age-specific hysterectomy prevalence. The influence of hysterectomy prevalence on uterine and ovarian cancer incidence rates and probabilities is illustrated.METHODS: Hysterectomy prevalence estimates are derived from cross-sectional data using a life table method. Analysis is based on hysterectomy data from the Utah Hospital Discharge Data Base and cancer cases recorded by the Utah Cancer Registry, 1995 1997.RESULTS: Correction for hysterectomy prevalence increases cancer incidence rates, more so at older ages, such that by ages 85 and older the increase is 92% for uterine cancers of the cervix and corpus, and 17% for ovarian cancer. Over the life span, correction for hysterectomy prevalence increases the estimated number per 100,000 developing these diseases by 195 (from 603 to 798) for uterine cervical cancer, 1,553 (from 2,580 to 4,133) for corpus uterine cancer, and 911 (1,674 to 2,585) for ovarian cancer.CONCLUSIONS: The utility of the cross sectional method for deriving hysterectomy prevalence is that it does not require several years of data to obtain reasonable estimates. Correction for hysterectomy prevalence has a large effect on the cancer rates and probabilities, providing a more accurate representation of the risk and burden of these cancers. PMID- 11018385 TI - Complex ovarian cysts in postmenopausal women are not associated with ovarian cancer risk factors. Preliminary data from the plco cancer screening trial. AB - PURPOSE: We assessed whether ovarian abnormalities detected on ultrasound in postmenopausal women are precursors to ovarian cancer.METHODS: We compared the transvaginal ultrasound findings from the initial examination of twenty thousand postmenopausal women enrolled to date in an ongoing randomized trial of cancer screening to data on the established risk factors for ovarian cancer obtained from self-administered questionnaires. We distinguished cysts with the suspicious characteristics of a septum, solid component, irregular or thick wall ("complex cysts") from simple sonolucent cysts with none of those features.RESULTS: High parity, protective for cancer, was negatively associated with complex cysts (Odds Ratio ["OR"] for five or more births versus no births = 0.72, 95% CI = 0.53 0.97), but long-term oral contraceptive use was not (OR = 0.96, 95% CI = 0.76 1.20). A family history of ovarian cancer or multiple breast cancers, a strong risk factor for cancer, was not associated with complex cysts (OR = 0.99, 95% CI = 0.68-1.44). Other abnormalities found on ultrasound (including simple cysts, bilateral cysts, or all abnormalities combined) also did not share the established risk factors for ovarian malignancy. We formed no combination of features of abnormalities (septum, echogenicity, size, or papillary projection) with the cancer risk factor profile.CONCLUSIONS: Although a very small proportion of the clinically silent ovarian abnormalities found on ultrasound are found to be ovarian cancers, the remaining complex cysts and other clinically suspicious abnormalities do not appear to be the immediate precursors of ovarian cancer. PMID- 11018386 TI - Use of hormone replacement therapy (Hrt) And detection of human papillomavirus (Hpv) Dna in postmenopausal women. AB - PURPOSE: This study investigated the association between detection of HPV DNA among current and past HRT users compared to never HRT users, duration of HRT use, and type of HRT (estrogen, estrogen/progestin).METHODS: Postmenopausal women (n = 390) were recruited from a university hospital and completed a questionnaire regarding 1) HRT use, duration, and type, 2) reproductive and sexual history, 3) smoking and alcohol use, and 4) HPV-related diseases. Cervical specimens were obtained for Pap smears, and for the presence of HPV using PCR/dot blot and DNA sequencing, and Southern blot or SSCP. Age-adjusted odds ratios (OR), 95% confidence intervals (CI), and logistic regression examined the association between HRT use and risk of HPV detection.RESULTS: The frequency of HPV was 10%, with 2.8% oncogenic types. Compared to Never Users, Current (adj. OR = 1.50, 95% CI = 0.55, 4.07) and Past (OR = 1.96, CI = 0.56, 6.86) HRT users had an elevated risk of HPV detection. Although HRT duration among Current Users was not statistically significant (OR = 1.01, 95% CI = 0.95, 1.07), duration among Past Users was significantly associated with an increased risk of HPV detection (OR = 1.30, CI = 1.06, 1.61). In addition, Past Users of estrogen/cyclic progestin HRT regimens were significantly more likely to be detected with HPV (OR = 1.82, CI = 1.11, 2.97) compared to Never Users.CONCLUSIONS: These findings indicate that risk of HPV detection is increased among users of HRTs, particularly among Past Users associated with longer duration and among those taking HRT regimens that included a progestin. Results suggest a latency or duration effect may be important in elevating the risk. They also support previous in vitro studies of progestin effects on HPV gene regulation. PMID- 11018387 TI - Second primary ovarian cancer among women with cancer. AB - PURPOSE: Some studies suggest women with certain types of cancers are at increased risk for ovarian cancer. This study assessed the risk of second primary ovarian cancer among U.S. women who have cancer by anatomic site, age, race, and time since diagnosis of the first primary cancer.METHODS: We analyzed data from SEER cancer registries for women diagnosed with invasive cancer between 1973 and 1996. Person-years were accumulated from 2 months after initial cancer diagnosis to date of ovarian cancer diagnosis, death, loss to follow-up, or end of follow up, December 31, 1996. The expected number of cases was obtained by multiplying 5 year age and calendar year interval specific ovarian cancer rates by the accumulated person-years at risk. We calculated the risk (observed [O]/expected numbers [E]) of second primary ovarian cancer by cancer site and age (<50 years, >/=50 years), race (all, white, black), and time since first cancer (0-4, 5-9, 10 14, 15-24 years). Statistical tests and 95% confidence intervals (CI) were based on the assumption of a Poisson distribution.RESULTS: A significant increased risk of ovarian cancer was found for women aged <50 years at time of diagnosis with melanoma (O/E = 3.5, 95% CI = 2.1-5.5) and cancer of the breast (O/E = 6.0, 95% CI = 4.9-7.2), cervix (O/E = 4.2, 95% CI = 2.6-6.3), corpus uteri (O/E = 11.91, 95% CI = 7.3-18.4), colon (O/E = 17.9, 95% CI = 11.1-27.3), and ovary (O/E = 4.9, 95% CI = 2.7-8.2); no increased risk was found for women aged >/=50 years. Ovarian cancer risk remained elevated following all of these first primary cancers 5-9 years after diagnosis; for women with breast and colon cancer, risk remained elevated 15-24 years after diagnosis. A significant increased risk was found for all of these cancers among white women <50 years at diagnosis; risk was increased among black women <50 years with cancer of the breast, cervix, and colon.CONCLUSIONS: We found an ovarian cancer risk higher than expected for women with certain types of cancer; however, the risk was limited to women <50 years of age. PMID- 11018388 TI - Pap smear screening in an equal-access health care system. Yield of screening and predictors of squamous intraepithelial lesions and atypical squamous cells of undetermined significance. AB - PURPOSE: The purpose of this study was to examine predictors of squamous intraepithelial lesions (SIL) and atypical squamous cells of undetermined significance (ASCUS) in an equal access health care setting.METHODS: A cohort of 10,713 active-duty United States Air Force women who provided one cervical/vaginal smear (CVS) as part of routine medical care between January 1, 1994 and December 31, 1996 were selected for this study. Each of the specimens was characterized by a "Bethesda System" diagnosis: within normal limits, repair/reactive, ASCUS, low grade SIL, high grade SIL, invasive carcinoma, infectious disease or unsatisfactory.RESULTS: Significant disease was associated with age younger than 30 years, enlisted rank, Hispanic ethnicity and the presence of genital warts on examination. SIL was associated with age 21-25 and Hispanic ethnicity in the logistic regression model. A diagnosis of ASCUS was associated with age younger than 30, marital status as single/never married, presence of genital warts on physical examination, and a history of previous abnormal CVS since entering military service. Age younger than 30, a marital status of single/never married and a previous abnormal CVS were associated with ASCUS in the logistic regression model.CONCLUSIONS: The disparity noted between women of low and high socioeconomic status was not seen here; those with the lowest income and education were not significantly more likely to have SIL or ASCUS when age was considered in the model. The significant association between Hispanic ethnicity and SIL, which should be further explored, may indicate the need for increased screening in this population. PMID- 11018389 TI - Comparison between hybrid capture II and polymerase chain reaction results among women at low risk for cervical cancer. AB - PURPOSE: Hybrid Capture (HC) is a new and powerful tool to detect DNA/RNA of several infectious agents. Regarding HPV DNA detection HC II has shown no false positive results due to contamination and its inter-laboratory reproducibility reaches 98%. It is also simpler and costs less than other methods such as Polymerase Chain Reaction (PCR) and it has shown a high correlation to clinical findings. Our objective is to compare HC II and PCR results among women at low risk for cervical cancer.METHODS: This is a cross-sectional study which enrolled 977 asymptomatic women aged 15 and 70 years. Squamous columnar junction samples were obtained and analyzed for HPV DNA either by HC II and PCR. Epidemiological factors and cyto-pathologic results were related to HPV DNA status. The Pearson Chi-square test and multiple logistic regression were performed to relate epidemiological and cytopathological variables and HPV DNA status.RESULTS: About 15.4% and 15.9% of women were HPV DNA positive by HC II and PCR respectively. Both were highly associated with cytology (P < 0.0001). The correlation to histopathology was also good with a higher significance for HC II. Factors related to HPV DNA detection by PCR were: oral contraceptive use (OR = 1.26; 95% CI = 1.2-1.62); history of genital HPV infection (2.11; 1.13-4.59); 3 or more sexual partners (1.35; 1.10-1.83); education (/=45).Rates are similar between SRRHIS and SEER except in the case of black women over the age of 45. These women in rural SRRHIS have 1.66 times the incidence rate compared to those in nonrural SRRHIS and 1.97 times the rate of rural SEER.RESULTS:Table 1CONCLUSIONS: The results of the study suggest that the incidence of cervical cancer in the region is consistent with the mortality rates. The age-group with the highest rates are black women over 45 in rural areas, suggesting the need to target this group to reduce the racial disparity in cervical cancer. PMID- 11018391 TI - Incidence of squamous neoplasia of the cervix and vagina in des-exposed daughters. AB - PURPOSE: Women exposed to diethylstibestrol (DES) in utero are known to have an excess risk of clear cell adenocarcinoma of the vagina and cervix, in addition to vaginal epithelial changes, but the effect on the incidence of squamous neoplasia is uncertain. This study evaluated the long-term risk of developing high-grade squamous neoplasia of the genital tract among women exposed prenatally to diethylstilbestrol.METHODS: A cohort comprising 3899 DES-exposed and 1374 unexposed daughters was followed for thirteen years (1982-1995) for pathology confirmed diagnoses of high-grade squamous neoplasia. A pathologist blinded to exposure status reviewed seventy-seven percent of cases. Poisson regression analysis was used to compute relative risks (RR) and 95% confidence intervals (CI) controlling for age, calendar year, screening history and other covariates.RESULTS: The RR (95% CI) among DES-exposed versus unexposed, based on 111 cases of high-grade disease, was 2.12 (1.19-3.77). Adjustment for screening history had little effect, but when the analysis was restricted to a group highly screened before 1982, the risk was reduced. Risk estimates were higher among women exposed earlier in gestation; the RR (95% CI) for exposure within 7 weeks of the last menstrual period was 2.82 (1.43-5.53).CONCLUSIONS: The findings support an association between in utero DES exposure and high-grade squamous neoplasia, although a role for more intensive screening among DES-exposed women in the production of this excess could not be completely ruled out. PMID- 11018392 TI - Use of topical sunscreen and the risk of malignant melanoma. Results of a meta analysis of 9,067 patients. AB - PURPOSE: Prior epidemiological studies suggest that the use of sunscreen preparations is associated with increased risk of cutaneous malignant melanoma (CMM) although the data are conflicting. A meta-analysis was performed to evaluate this suspected association.METHODS: A protocol was developed for a meta analysis examining the association between frequent sunscreen use versus non-use and the development of CMM. Data from observational studies were pooled using a general variance based meta-analytic method employing confidence intervals. The outcome of interest was a summary relative risk (RRs) reflecting the risk of melanoma associated with sunscreen use versus non-use. Sensitivity analyses were performed when necessary to explain any observed statistical heterogeneity.RESULTS: Eleven case-control studies enrolling 9,067 patients were combined in a meta-analysis. This yielded a RRs of 1.11 (CI = 0.37-3.32), a non statistically significant result, (i.e. no association between sunscreen use and melanoma risk). Since the data were found to be heterogeneous, i.e. Q = 42.0 (p < 0.001), a series of sensitivity analyses were performed to explore possible sources of heterogeneity. Stratifying studies based on study design, i.e. hospital based versus population based, showed that hospital derived data were highly heterogeneous (Q = 36.9, p < 0.001) while the population registry data were not (Q = 4.9, p = 0.18). Combining those studies using population based data gave a RRs of 1.01 (95% CI = 0.46-2.28) indicating no association between sunscreen use and the development of CMM.CONCLUSIONS: The available epidemiological data do not support the existence of a relationship between topical sunscreen use and an increased risk of CMM. PMID- 11018393 TI - Self-report of smoking, obesity and hypertension history and survival among a cohort of iowa renal cell carcinoma cases. AB - PURPOSE: Smoking, obesity and hypertension are well-established risk factors for the development of renal cell carcinoma (RCC). However, little is known regarding the ability of RCC patient history regarding these characteristics to predict future survival.METHODS: To evaluate this question, we conducted follow-up on a cohort of incident RCC cases first assembled in Iowa from 1986-1989 for case control analysis. We report data on 364 cases aged 40-86 years who reported detailed anthropometric and lifestyle history on a mailed questionnaire. Mortality experiences through 1998 were determined by linkage to the Iowa Cancer Registry and other databases; 233 deaths were identified during the 2,470 total years of follow-up, with 145 of those having RCC listed as the underlying cause of death on the death certificate. The Cox Proportional Hazards model was used to estimate the risk associated with each potential prognostic characteristic.RESULTS: After adjustment for age, a suggestion of an association with survival was noted for history of hypertension [Relative Risk (RR) = 0.8; 95% Confidence Interval (CI) 0.5-1.1] but no association was noted for either smoking or weight history. After further adjustment for tumor stage at diagnosis, evidence of an association for either smoking or obesity remained absent. However, after similar adjustment, the protective effect of history of hypertension actually strengthened slightly (RR = 0.7; CI 0.5-0.9).CONCLUSIONS: Data from this follow-up study of incident RCC cases in Iowa suggests that after adjustment for the strongest independent predictor of survival, a reported history of hypertension is associated with better survival. PMID- 11018394 TI - The excess incidence of squamous cell esophageal cancer among us black men. Role of social class and other risk factors. AB - PURPOSE: To investigate the relationship between social class factors and squamous cell esophageal cancer and the extent to which alcohol, tobacco, diet, and social class contribute to the five-fold higher incidence among black than white men in the United States.METHODS: Interviews were conducted with 347 incident cases of squamous cell esophageal cancer (119 white males and 228 black males) and 1354 population-based controls (743 white males and 611 black males) from Atlanta, Detroit, and New Jersey. Risks were estimated using unconditional logistic regression controlling for potential confounders.RESULTS: Elevated risks of squamous cell esophageal cancer were associated with indicators of low social class, especially low annual income. The adjusted odds ratios (ORs) for subjects with incomes < $10,000 versus incomes of $25,000 or more were 4.3 (95% CI = 2.1 8.7) for whites and 8.0 (95% CI = 4.3-15.0) for blacks. The combination of all four major risk factors: annual income less than $25,000, moderate/heavy use of alcohol, use of tobacco for six months or longer, and consumption of less than 2.5 servings of raw fruits and vegetables per day accounted for almost all of the squamous cell esophageal cancers in whites (98%) and blacks (99%), and for 99% of the excess incidence among black men.CONCLUSIONS: Lifestyle modifications, especially a lower intake of alcoholic beverages, would markedly decrease the incidence of this cancer in both races and narrow the racial disparity in risk. Further studies into the determinants of social class may help identify a new set of exposures for this tumor that are amendable to intervention. PMID- 11018395 TI - Association of dietary antioxidants on the severity of gastritis in a high risk population. AB - PURPOSE: This study is to examine the role of dietary antioxidant intake on the severity of gastritis at the baseline in an intervention study, Chemoprevention of Gastric Dysplasia.METHODS: The Chemoprevention of Gastric Dysplasia is an intervention study conducted in the towns of Pasto and Tuquerres, Colombia. A 79 item food frequency questionnaire was administered to 1,219 subjects at baseline. Endoscopic screening for precancerous gastric lesions was performed in all subjects by biopsy and histologic diagnosis. Subjects without adequate diagnostic material (n = 18) and subjects identified as histologically normal (n = 12) or with gastric cancer (n = 4) were excluded from these analyses. Analysis of Variance on logarithm-transformed data was conducted on dietary antioxidants and the stages of gastritis (atrophic gastritis, intestinal metaplasia, and gastric dysplasia).RESULTS: After adjusting for age, body mass index, smoking status, year of education, total calorie intake, and H-pylori infection status, dietary vitamin C and vitamin E were found to be inversely associated with the severity of gastritis (p < 0.05). Decreased dietary alpha-carotene was found to be marginally associated with gastric dysplasia. Data were then stratified by town where subjects were recruited. The trends for the observed associations were apparent for dietary alpha-carotene, vitamin C and E. However, the association was significant only for vitamin C in Pasto. Compared to subjects with atrophic gastritis, subjects diagnosed with gastric dysplasia have an approximately 15% lower intake of vitamin C.CONCLUSIONS: This study confirmed the protective effect of specific dietary antioxidants on the severity of gastritis. Dietary vitamin C, and to a lesser extent, dietary vitamin E are potentially important for the prevention of gastric cancer. PMID- 11018396 TI - Using north carolina medicare data to assess excess prostate cancer mortality among african americans. AB - PURPOSE: To investigate the basis for the higher prostate cancer mortality rate for African American (AA) men, which is twice the rate for White men.METHODS: 221 AA and 979 White men with a primary diagnosis code of prostate cancer ("patients") in the North Carolina Medicare Hospitalization claims from 1997 were compared with 1,326 AA and 5,874 White men of the same age with no cancer hospitalizations ("beneficiaries") selected from the NC Medicare Enrollment files. Mortality rates were calculated as the cumulative percent of deaths using the hospital discharge date as day 1. AA and White age distributions were similar.RESULTS: Cumulative mortality percentages at 6, 12, and 18 months were, respectively, 4.5, 7.7, 10.9 for AA patients; 2.8, 6.5, 9.2 for White patients; 2.3, 3.8, 7.4 for AA beneficiaries; and 1.8, 3.1, 6.1 for White beneficiaries.CONCLUSIONS: AA prostate cancer patients had higher overall mortality than did White prostate cancer patients during the first year, but by 12-months the White-Black survival advantage for prostate cancer patients was similar in magnitude to the White-Black survival advantage among the non-cancer Medicare beneficiaries. AAs' higher prostate cancer mortality may derive from higher short-term case fatality rates, which may reflect differences in treatment and access to quality medical care, co-morbidities, and tumor characteristics such as stage and grade at diagnosis, and in part from the survival disadvantage for AA in the general population. PMID- 11018397 TI - Lung cancer and indoor air pollution in rural china. AB - PURPOSE: Indoor air pollution has been linked with lung cancer in China. In contrast to previous studies conducted in urban areas with high levels of industrial pollution, we undertook a lung cancer case-control study in a rural area of China, where residents live in underground dwellings. We evaluated the effects of radon, wood and coal combustion, cooking oil fumes, and environmental tobacco smoke on lung cancer risk.METHODS: We enrolled 886 lung cancer cases (656 males, 230 females) diagnosed between 1994-98, aged 30-75 years and 1765 frequency matched population-based controls from two prefectures in Gansu Province in Northwestern China. We conducted interviews with subjects or next of kin on smoking, housing characteristics, fuel use and cooking practices. Year long radon detectors were placed in current and former homes of subjects.RESULTS: Subjects primarily used coal (22%), wood (56%) or a combination of both (22%) for heating. Odds Ratios (OR) for lung cancer rose with increasing percent of time that coal was used to heat homes over the past 30 years (ORs = 1.00, 1.17, 1.35, 1.23 compared to wood only, adjusted to smoking, P for trend = 0.025). Among non smoking females and males, the OR for ever exposed to environmental tobacco smoke was 1.19, 95% CI = 0.7-2.0 with a significant trend for increasing years of exposure. Fumes from cooking with rapeseed oil increased the risk of lung cancer (OR = 1.56, 95% CI = 1.0-2.5) among non-smoking women. Among these women, occasional and frequent eye and throat irritation during cooking appeared to be associated with increased risk of lung cancer (ORs = 1.00, 1.42, 2.28, p trend < 0.01), whereas, increasing level of smokiness during cooking did not appear to affect risk.CONCLUSIONS: There is a suggestion that coal used for heating, environmental tobacco smoke, and cooking oil fumes contribute to the risk of lung cancer in this rural area of China. PMID- 11018398 TI - Trends in colorectal cancer rates in urban shanghai, 1972-1996, in relation to dietary changes. AB - PURPOSE: In urban Shanghai, the largest industrial and commercial city in China, the age-adjusted (world standard) incidence rates for colorectal cancer increased from 14.5 to 23.3 per 10(5) men and from 12.1 to 20.3 per 10(5) women between 1972 and 1996. This change was even more pronounced for colon cancer, whose incidence rates doubled from 5.95 to 13.7 per 10(5) men and from 5.77 to 12.5 per 10(5) women. The reasons for the rapid increases in cancer rates are not fully understood, but may involve dietary exposures that have changed substantially over the past two decades.METHODS: We calculated Pearson correlation coefficients (r) between colorectal cancer rates and the dietary factors of grain, vegetable oil, pork, poultry and vegetable consumption over the period of 1972 through 1996 in urban Shanghai.RESULTS: Statistically significant positive associations were observed between colon cancer rates and per capita consumption of vegetable oil (r = 0.91 for men, r = 0.94 for women), poultry (r = 0.90 for men, r = 0.90 for women), and pork (r = 0.78 for men, r = 0.81 for women). The correlation coefficients were not statistically significant between colon cancer and per capita consumption of grain (r = 0.38 for men, r = 0.37 for women) or vegetables (r = 0.16 for men, r = 0.14 for women). Similar weaker associations were observed between rectal cancer rates and vegetable oil, pork and poultry consumption.CONCLUSIONS: The findings in our study suggest that increases in dietary fat, poultry and pork intake may play a role in the rising colorectal cancer rates in Shanghai. PMID- 11018399 TI - Prostate cancer risk. Medical history, sexual, and hormonal factors. AB - PURPOSE: Various medical conditions, infectious agents, sexual, and hormonal factors have been investigated in relation to prostate cancer risk. Given inconsistent results these factors were examined in this study.METHODS: This population-based case-control study was conducted in northeastern Ontario from 1995 to 1999. Cases (n = 760), aged 45 to 84 at the time of diagnosis, were identified through the Ontario Cancer Registry and diagnosed between January 1995 and December 1998. Controls (n = 1,634) were age-frequency matched and were selected from the northeastern Ontario population using published telephone listings. Mail and telephone questionnaires were used for data collection. Logistic regression was used to investigate risk associated with: 1) particular medical conditions and 2) hormonal and sexual factors. Cases were subdivided into those with symptoms of prostate disease and those with few or no such symptoms.RESULTS: Symptomatic cases who reported a history of venereal disease (age-adjusted odds ratio (OR) = 2.11, 95% confidence interval (CI) 1.18-3.80) and vasectomy (age-adjusted OR = 1.49, 95% CI 1.14-1.95) were at significantly increased risk of prostate cancer. Asymptomatic cases who reported a check-up at least once a year were at increased risk (age-adjusted OR = 1.46, 95% CI 1.08 1.98). Asymptomatic and symptomatic cases who reported a history of prostate cancer in a first degree relative were at increased risk (age-adjusted OR = 2.41, 95% CI 1.64-3.54; age-adjusted OR = 3.18, 95% CI 2.28-4.45, respectively). Symptomatic cases with a history of urinary tract infection were at non significantly increased risk (age-adjusted OR = 1.31, 95% CI 0.98-1.76). Heart disease, mumps, allergies, and height were generally not associated with prostate cancer.CONCLUSIONS: A history of venereal disease, family history of prostate cancer, and vasectomy were positively associated with prostate cancer. Further investigation of selected medical conditions, sexual, and hormonal factors in prostate cancer development is warranted. PMID- 11018400 TI - Geographic variation in prostate cancer mortality rates among white males in the united states. AB - PURPOSE: The most recent atlas of cancer mortality in the United States revealed elevated prostate cancer mortality rates among white males in the northwest, Rocky Mountain, northcentral, and southeast areas, as well as New England, especially during the 1970-94 period. We wanted to test whether this observed geographic variation was simply due to chance or not.METHODS: We used a spatial scan statistic using mortality data for 506 state economic areas.RESULTS: There were four significant clusters with elevated risks of prostate cancer mortality (P < 0.001). The most prominent cluster was in the northwestern quadrant of the country, followed by clusters in New England, the midwest, and southeast regions. Within the northwestern cluster, we also detected seven significant sub-clusters (P < 0.05).CONCLUSIONS: We concluded that the observed geographic variation of prostate cancer mortality is indeed real, and deserves further study into the underlying determinants. PMID- 11018401 TI - Distinct distribution of rare us kshv genotypes in south texas. Implications for kshv epidemiology and evolution. AB - PURPOSE: Kaposi's sarcoma-associated herpesvirus (KSHV, or human herpesvirus 8) is etiologically associated with Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). Although previous studies have assessed the geographic distribution of KSHV genotypes, the molecular epidemiology of KSHV remains largely unknown. The purpose of the present study was to examine the genotypes of KSHV isolates from KS patients in South Texas.METHODS: Eighteen KSHV isolates from 16 KS and 1 PEL patients in South Texas were collected between 1996 and 1998 and analyzed for KSHV subtypes by PCR sequencing of ORFK1 gene and KS330 fragment, and by PCR of ORFK15 gene. DNA sequences were aligned with known sequences and KSHV subtypes were assigned based on sequence variations.RESULTS: Of 18 KSHV isolates, 13 exhibited C subtype, and 5 exhibited A subtype in ORF K1 gene. ORF K15 genotyping showed that 10 of the isolates exhibited M form, of which 9 had C3 subtype. A unique C subtype isolate was found and classified as C6 clade. All of the M form KSHV isolates were found among KS patients over 50 years of age. Conversely, all KS patients under 40 years of age had only the P form KSHV isolates.CONCLUSIONS: In South Texas there is a distinct distribution of C3/M KSHV isolates, which are rarely found in other US regions (1 of 29). The C3/M KSHV genotype is more prevalent in HIV-negative elderly KS patients while the P-form of KSHV is more common among many young AIDS-KS patients. PMID- 11018402 TI - Possible relationship between birth weight and cancer incidence among young adults. AB - PURPOSE: Fetal and early life events have been associated with diseases that develop later in life. Low birth weight and the adult onset of hypertension, coronary heart disease, cerebrovascular disease, and non-insulin dependent diabetes have been identified. As well, associations with breast, ovarian, and prostate cancer to high birth weight have been found. An assessment of birth weight and cancer incidence was conducted in a cohort of black and white residents under the age of 46 years.METHODS: Cases were obtained from the Savannah River Region Health Information System cancer registry incident cases (1991-1995) and were limited to South Carolinians born in 1950 and later. Controls were obtained from birth certificate records by choosing the next two records after a cancer case record that matched on year of birth, race, and sex. Results were obtained for 117 cancer cases and 238 controls.RESULTS: After examining the birth distribution, the births were split into two groups based on mean birth weight among controls (3215 grams). Conditional logistic regression (CLR) showed that individuals with higher birth weights (> = 3215 g) were 1.65 (95% CI = 1.03-2.64) times more likely to be cancer cases than those with lower birth weights. When weights were categorized into 500 g increments, a CLR Score statistic showed there was a significant trend (p = 0.0006) of increasing proportion of cancer cases with increasing birth weight. Eight out of the eight cases of lymphoma had birth weights greater than 3579 g.CONCLUSIONS: The results of this preliminary study suggest that cancer incidence among the young may be associated with higher birth weights. One possible reason for this finding, which requires further investigation, might be that larger infants are exposed to higher levels of hormones and/or growth factor than smaller infants in utero that might increase the risk of certain cancers later in life. This may be suggestive of possible environmental factors affecting early growth. These findings support the need for additional study of this association. PMID- 11018403 TI - An epidemic without illness. Using dna markers to model infection. AB - PURPOSE: Combining molecular biology with infection control interventions can increase compliance and allow objective measurement of effectiveness. We developed a group of PCR detectable non-infectious DNA markers that can be used to model infection and provide immediate feedback on hygiene practices in institutional settings. In previous studies, we illustrated that the markers were spread in the environment in the same manner as infectious particles.METHODS: We are conducting a prospective study in 10 child care centers in order to 1) confirm that the DNA markers are valid surrogates for bacteria and viruses; 2) identify specific foci of contamination and modes of transmission; 3) illustrate the effectiveness of infection control programs utilizing the DNA markers. Centers are randomized to receive an interactive educational infection control intervention or a standard immunization intervention. The DNA markers are introduced into the center and the rate of dispersion of the DNA markers is compared with directly observed changes in hygiene behavior among the staff.RESULTS: Initial results indicate that the markers can be removed mechanically by hand washing and that common over-the-counter cleaners are effective in inactivating the markers. Toys, countertops and doorknobs appear to be more important as infectious reservoirs than brief casual contact. Data from the prospective study will be available prior to September, 2000.CONCLUSIONS: This novel approach utilizing an objective measurement will be used to identify the interaction between behavior and environmental reservoirs of infection and drive future strategies for infection control. PMID- 11018404 TI - 'of Mice and men' (John steinbeck)-How do we determine the potential for immunotoxicity in humans? AB - PURPOSE: Immunotoxicology is most simply defined as the study of adverse effects on the immune system resulting from exposure to drugs, environmental and industrial chemicals, and in some instances, biological materials. The science of immunotoxicology has validated animal models to conduct risk assessment. However, approaches to characterize immunotoxicity in humans are poorly defined. Animal models have indicated that a primary immune response is most predictive of immunotoxicity. Because vaccines can trigger a primary immune response, this approach may have utility in humans. The purpose of this project was to determine if the response to the influenza vaccine can be validated as an objective measurement of immune status in the workplace.METHODS: We randomly selected employees to receive the influenza vaccine and employees, matched according to age and gender, to receive the placebo. The participants (32 test group and 19 placebo group) completed a brief questionnaire to identify potential confounding factors. Specific anti-influenza antibodies were measured in the serum via an ELISA 30 days after administering the vaccine or placebo.RESULTS: Only 50% (16 of 32 subjects) produced a positive response which is defined by the Centers for Disease Control as a four-fold increase in serum titers. Not unexpectedly, all samples contained antibody to influenza prior to vaccination, and a number of the participants who did not achieve a positive response started with high serum titers.CONCLUSIONS: The influenza vaccine was originally selected for this study because of costs, acceptability to the population and proven safety and efficacy. However, the results of the present investigation suggest that this approach will have little utility as a workplace monitor of human immunotoxicity because most individuals would not be making a primary antibody response. Other technical limitations with the influenza vaccine will be presented; and the alternatives for other biomonitors of human immunotoxicity will be discussed. PMID- 11018405 TI - Analyzing sensitivity to model form assumptions of infection transmission system models. AB - PURPOSE: Transmission system models make restrictive assumptions that might distort the conclusions of model analyses. We propose methods to progressively relax the following assumptions of classical deterministic compartmental models: 1) that the population has an effectively infinite size 2) that contact is instantaneous with no duration, 3) that mixing in this large population is instantaneously thorough after contact.METHODS: Analyses of contact patterns between high and low risk groups on gonorrhea transmission were performed. Initial models were similar to those analyzed by Hethcote and Yorke with compartments corresponding to sets of individuals. The instantaneous contact assumption in these models was relaxed by using continuous deterministic pairing models in the style of models presented by Dietz and Hadelar. That model makes restrictive assumptions about concurrent contacts, population sizes, and instantaneously random mixing. To relax these assumptions, we simulated our GERMS model of discrete individuals forming pairings and transmitting infection in continuous time.RESULTS: Relaxing the instantaneous contact assumption demonstrated a progressively decreased effect of mixing between high and low risk groups as the duration of contact was increased. The GERMS model simulations were shown to effectively reproduce pairing model behavior given the same restrictive assumptions as the pairing model. Further GERMS model analysis then demonstrated that concurrency assumptions alter the effects of contact rates between risk groups in ways that are dependent upon contact parameters. Finally GERMS models were used to structure mixing into four local areas. This affected the dynamics of reaching equilibrium but not the equilibrium value.CONCLUSIONS: Assessing the effects of assumptions in continuous compartmental models of transmission systems is feasible and important. PMID- 11018406 TI - The multiperson use of non-syringe injection equipment and risk of hepatitis c infection in a cohort of young adult injection drug users, chicago 1997-1999. AB - PURPOSE: The possibility that hepatitis C virus (HCV) is transmitted via the multiperson use of injection paraphernalia other than syringes has been suggested, but epidemiologic studies to examine the association are difficult to design due to saturation levels of infection in most samples of injection drug users (IDUs). This study (1) assembled a sample of young adult IDUs, among whom hepatitis C infection prevalence was still moderate, (2) measured incident HCV infection, and (3) determined the risk for seroconversion associated with specific forms of sharing injection paraphernalia.METHODS: Between 1997 and 1999, 702 IDUs, 18-30 years old, were interviewed and screened for antibodies to HCV at baseline and at 6 and 12 months post-baseline. Participants were recruited through street outreach, advertising and chain-referral from ethnically diverse neighborhoods in metropolitan Chicago. Data were analyzed using standard survival statistical methods.RESULTS: HCV prevalence was 27% at enrollment. During 296.5 person-years of observation, we observed 37 HCV seroconversions (incidence: 12.5/100 person-years). The adjusted relative hazard (RH) of seroconversion, after controlling for demographic and drug use covariates, was highest for sharing cookers (RH, 3.58; 95% CI 1.47-8.70), followed by sharing rinse water (RH, 2.16; 95% CI 1.03-4.52), and cottons (RH, 1.84; 95% CI 0.95-3.56). Risk associated with syringe sharing was marginally significant, and backloading was unassociated with hepatitis C seroconversion. Adjusting for syringe sharing, the independent effects of sharing cookers and cottons remained significant, and a final model that included each sharing practice demonstrated that sharing cookers had at the strongest association with seroconversion (RH, 3.03; 95% CI 1.05 8.72).CONCLUSIONS: This study suggests that sharing non-syringe paraphernalia may be an important cause of hepatitis C virus transmission between IDUs. PMID- 11018407 TI - Prevalence and co-colonization with group b streptococcus (Gbs) Among heterosexual college couples. AB - PURPOSE: Maternal colonization with GBS is a major cause of neonatal sepsis and meningitis. While GBS transmission is assumed to be fecal-oral or person-to person direct contact including possible sexual transmission, risk factors for colonization and carriage are poorly described. Basic descriptive epidemiologic information is needed in order to implement a prevention program such as the development of an effective vaccine.METHODS: As part of a study of heterosexual transmission of urinary tract infection (UTI), we describe the prevalence and co colonization of GBS among college couples. Self-collected vaginal, urine and rectal specimens were cultured from women with UTI and their current sex partners ("case couples"), and from women without UTI and their current sex partners ("control couples").RESULTS: Thirty-four percent (98/285) of case women and 31% (56/182) of control women had at least one GBS isolate. 25% (45/177) of case men and 26% (32/122) of control men had at least one GBS isolate. Given that GBS was present in one partner, 45% of case (34/75) and 57% of control (24/42) couples were co-colonized. Using pulsed-field gel electrophoresis (PFGE) analysis, GBS isolates shared within couples were identical in most cases, but PFGE patterns differed across couples.CONCLUSIONS: Prevalence of GBS is high among sexually active college students and sexual contact is associated with increased transmission. PMID- 11018408 TI - A definition for chronic sequelae applied to campylobacter and guillian-barre syndrome (Gbs). AB - PURPOSE: Chronic sequelae (CS) are increasingly important to drinking water risk assessment and management processes, but there has been relatively little scientific rigor in defining CS or reviewing the literature on water-related CS. Our purposes were to develop a scientific definition for CS and to evaluate the definition's practical merit.METHODS: We examined scientific publications for definitions of "chronic sequela." We developed a definition that is based on scientific concepts and that can be systematically applied to literature to assess whether pathogen-related health outcomes qualify as CS. As a case study, we conducted an extensive Medline search and tested our definition on the epidemiological and clinical literature linking Campylobacter and GBS.RESULTS: We defined "chronic sequela" as the secondary adverse health outcome that 1) occurs as a result of a previous infection by a microbial pathogen, and 2) is clearly distinguishable from the health events that initially result from the causative infection, and 3) lasts 3 months or more after recognition. The 12 Campylobacter and GBS studies (five epidemiological and seven clinical) revealed that current data reporting practices limit the evaluation of all three elements in our definition. Laboratory methods and criteria to characterize infection were not always adequately reported. Primary and secondary health events were always reported, but eight of the studies required obtaining additional articles to determine the GBS criteria used. Ten of the 12 articles contained duration data for the GBS symptoms.CONCLUSIONS: Much of the evidence needed to apply our definition was found in the studies reviewed, but changes in reporting practices would facilitate the scientific evaluation of pathogen-CS relationships and estimation of their public health magnitude. PMID- 11018409 TI - Prevalence of and risk factors for helicobacter pylori infection in school children in mexico. AB - PURPOSE: This study examined the prevalence of Helicobacter pylori (H. pylori) infection in school children from an urban area in Northwest Mexico and attempted to identify the risk factors that predispose children to infection in the community.METHODS: The cross-sectional study was conducted in 1997/98 in the poorest socio-economic sectors of the city of Hermosillo, Sonora, among 178 children aged 9 and 10 years. H. pylori status was determined in children by the accurate and safe non-invasive 13C-urea breath test. Family socio demographic/socio-economic status and living conditions data were elicited from parents by interview via structured questionnaires and/or direct observation.RESULTS: The overall prevalence rate of H. pylori infection for the children in Hermosillo as determined by this study was 47.1%. The overall prevalence rate of H. pylori infection was not found to be related to either gender (p = 0.531) nor to the two age groups (p = 0.483). It was not related to child's birthplace (p = 0.291) even after controlling for age and gender. However, the overall prevalence of infection was strongly and significantly associated with the parent's birthplace; rural-born mother (p = 0.028) and rural born father (p = 0.029). There was a noticeable lack of statistical association with the presence of animals at home (p = 0.988) and with direct contact with indoor/outdoor animals for study children (p = 0.099). When all of the explanatory variables that were significant in the bivariate analysis were fitted into a direct logistic regression model, the same strong effect of father's birth place (rural setting), number of siblings (>/=3 per family), type of main water supply (one tap in the yard) and the sharing of bed by the study child were observed as potential risk factors for acquiring the infection.CONCLUSIONS: The findings indicate that high prevalence rate of H. pylori infection observed on study children seems to depend on factors related to poor living conditions, particularly (but not exclusively) number of children, ruralism (rural-born father), the sharing a bed in childhood and type of main water supply (one tap in the yard). PMID- 11018410 TI - Antiretroviral prescribing patterns in the texas prison system. AB - PURPOSE: Although prison inmates are reported to exhibit substantially elevated rates of HIV infection, little is known about HIV treatment patterns, particularly pharmacotherapy, in correctional institutions. The purpose of the present study, therefore, was to describe antiretroviral prescribing patterns in one of the nation's largest prison populations.METHODS: The study population consisted of all known (n = 2,360) HIV-infected inmates incarcerated in the Texas prison system in 1998. Information on medical conditions, sociodemographic factors, and pharmacotherapy was obtained from an institution-wide medical information system. Inmates who received more than one type of pharmacotherapy in 1998 were included in the appropriate number of categories.RESULTS: In 1998, 66.8 percent (95% CI = 64.0-69.4) of all HIV-infected inmates with CD4 counts below 500 were treated with highly active antiretroviral therapy (HAART); and 31.1 percent (95% CI = 29.3-33.0) were given no antiretroviral therapy. Logistic regression results showed that HAART treatment decreased monotonically as a function of patient CD4 count category.CONCLUSIONS: A substantial proportion of HIV-infected TDCJ inmates were placed on therapies that were not consistent with the generally recommended DHHS guidelines for their disease stage. It will be important to for future investigations to assess whether such patterns continue to exist among prison populations, and to assess the determinants of these patterns. PMID- 11018411 TI - Haemophilus influenzae type b vaccination and risk of childhood leukemia in a vaccine trial in finland. AB - PURPOSE: To compare the incidence of childhood leukemia among children who received multiple early vaccine doses versus a single late dose in the context of a clinical trial in Finland during 1986 and 1987.METHODS: The trial was offered to the parents of all 114,000 children born in Finland between October 1, 1985 and August 31, 1987; 98% of the eligible children were enrolled. The vaccine consisted of heat-sized capsular polysaccharide of Haemophilus influenzae type b (Hib) coupled to diptheria toxoid (PRP-D). Children with odd birth dates received four doses of the vaccine at the ages of 3, 4, 6, and 14 months; children with even birth dates received only one dose, at the age of 24 months. The numbers of boys and girls with even and odd birth dates during the window of eligibility who remained alive during the follow-up period were obtained from the Finnish Population Registry. Numbers of leukemia cases diagnosed through 1996 among children with even and odd birth dates during the window of eligibility were obtained from the Finnish Cancer Registry. Poisson regression analysis of leukaemia incidence rates was conducted using the number of cases as the outcome variable and the natural logarithm of number of children at risk as an offset term. For comparison of the early and late vaccination groups, a binary variable based on odd or even date of birth was used.RESULTS: Among the 114,000 subjects, a total of 77 cases of childhood leukaemia were diagnosed: 33 in the early vaccination group and 44 in the late-vaccination group. The incidence of childhood leukaemia was lower in the early-vaccination group (relative risk 0.72, 95% confidence interval 0.46-1.13) than in the late-vaccination group, but the difference did not reach statistical significance.CONCLUSIONS: Our results suggest that early vaccination against Hib may reduce the incidence of childhood leukaemia, but confirmatory studies are needed. PMID- 11018412 TI - The changing epidemic of pediatric hiv infection in romania. AB - PURPOSE: By the late 1980s, over 100,000 infants and children were living in public institutions in Romania. It was not uncommon for children in these facilities to receive one or more 'micro-transfusions' of blood, unscreened for HIV, as therapy for anemia or malnutrition. To assess the impact of pediatric HIV infection in Romania, the European country with the most pediatric cases, cross sectional and cohort studies were implemented in Constanta (the epi-center of pediatric HIV in Romania) in April 1999.METHODS: Demographic, clinical and social data are collected once for all cross-sectional subjects. Similar data are collected every 11-13 months for subjects in the cohort. The cross-sectional study population was defined as all living HIV-infected infants and children, 0 18 years, known to the investigators from April to September 1999. The cohort consists of subjects diagnosed with HIV between 1995 and 1999.RESULTS: Enrolled are 791 subjects, of which 357 are in the cohort study. The majority (83%) are Romanian, vs Gypsy or Turkish/Hun and their mean age is 11 years (SD = 1.3). Biologic parents are the primary caretakers of 77% and 86% attend school. Mode of transmission was perinatal for 8%; blood transfusion/parental therapy modes account for 89% of the transmission and the presumed timing was between 1-12 months of age. Mean age at HIV confirmation was 5 years (SD = 3.2). AIDS has been diagnosed in 40% and 52% are receiving antiretroviral therapy.CONCLUSIONS: The Romanian pediatric HIV epidemic differs vastly from that in the US. Fewer children are with their biologic parent(s) and attending school. Early diagnosis of infection is rare, as therapy did not become available until the late 1990's. Perinatal transmission is increasing, however, which supports the need for HIV education and intervention in Eastern Europe. PMID- 11018413 TI - Prepregnancy body mass index and gestational weight gain as risk factors for preeclampsia and transient hypertension. AB - PURPOSE: Recent studies suggest prepregnancy obesity is a risk factor for preeclampsia, although only a handful of studies have examined the effect of gestational weight gain. The authors analyzed the effect of prepregnancy body mass index (BMI) and weight gain during pregnancy on risk of preeclampsia and transient hypertension.METHODS: Subjects were participants in a prospective cohort study of women who received prenatal care from thirteen obstetric practices in southern Connecticut (4/88-12/91). The women were interviewed in person before 16 weeks gestation and in the immediate postpartum period. All subjects' hospital delivery charts were abstracted. BMI was categorized as: <19.8 (underweight), 19.8-26 (normal: referent), 26-29 (overweight), >29 (obese). A gestational weight gain index, created using multiple linear regression, compared observed weight gain to the weight gain expected after adjustment for significant covariables (e.g. gestational aged at delivery). Logistic regression was used to estimate risk of preeclampsia (N = 44) and transient hypertension (N = 172) associated with prepregnancy BMI and gestational weight gain.RESULTS: Obese women had a mild increased risk of preeclampsia (OR = 1.81; 0.73-4.52); women in the other BMI categories had risks similar to that of normal BMI subjects. In contrast, risk of transient hypertension was substantially decreased among underweight women (OR = 0.35; 0.14-0.87) and substantially increased among obese women (OR = 3.43; 2.27-5.21). Higher than expected gestational weight gain did not increase the risk of preeclampsia. In contrast, risk of transient hypertension was increased over twofold among women in the highest quartile of the weight gain index (OR = 2.55; 1.66-3.92).CONCLUSIONS: Obesity appears to be a strong risk factor for transient hypertension and a milder risk factor for preeclampsia. High gestational weight gain was associated with increased risk of transient hypertension but not preeclampsia. PMID- 11018414 TI - Controlling for the effects of hospital attributes. The efficacy of factor analysis. AB - PURPOSE: The assessment of risk often requires controlling for the potentially confounding effects of hospital attributes. This paper (1) describes the use of factor analysis as a means of quantifying hospitals' obstetrical care, and (2) compares within respective unconditional logistic regression models the performance of factor scores with that of the "obstetrical unit service level" (OBUSL) classification as defined by the American Hospital Association (AHA).METHODS: A principal components factor analysis was performed on fourteen variables from 116 hospitals in Upstate New York. These variables, descriptive of hospitals' obstetrical care, were obtained from the 1992 AHA's Annual Survey of Hospitals. Factor scores were correlated with the OBUSL. Factor scores were matched to 89,341 women with vaginal deliveries in the 1992 Live Birth Registry for Upstate New York. The performance of factor scores and the OBUSL variable was compared in separate unconditional logistic regression analysis designed to identify determinants of obstetrical anesthesia care.RESULTS: Principal components factor analysis with varimax rotation identified three factors which were strongly correlated with hospital OBUSL. In a model which included the OBUSL variable, the adjusted odds ratios (AORs) or receiving an epidural for vaginal delivery were lower among mothers with Medicaid, HMO, or no insurance coverage (i.e., 0.45 [95% CI, 0.43-0.48], 0.68 [0.64-0.71], and 0.44 [0.38-0.52], respectively) than among those with private coverage. In a model in which factor scores were substituted for the OBUSL variable, respective AORs were 0.48 (0.45 0.52), 0.63 (0.60-0.66), and 0.45 (0.39-0.53).CONCLUSIONS: Factor analysis provided a parsimonious description of 14 hospital variables, was useful as a control within the regression model, and may prove similarly useful in other areas of clinical care. PMID- 11018415 TI - Intergenerational and environmental factors influencing pregnancy outcomes. AB - PURPOSE: Factors that relate to the mother's own course of growth and development might influence later reproductive performances. This study examines the effect of maternal birth weight and maternal height and maternal weight gain, on term small for gestational age (SGA) and preterm delivery.METHODS: Data used for this study were from the South Carolina Pregnancy Risk Assessment Monitoring System (PRAMS) and birth certificate files for 1993-1995. Since PRAMS data were weighted to adjust for sampling probabilities, non-response and survey design, Survey Data Analysis (SUDAAN) software was used in all analyses.RESULTS: Maternal birth weight was strongly associated with term SGA (OR = 3.65; 95% CI = 1.83-7.29), but not preterm delivery among black mothers; it was associated with preterm delivery (OR = 1.86; 95% CI = 1.09-3.17), but not term-SGA for white mothers. Maternal birth weight was associated with term SGA among nonsmokers only (OR = 2.34; 95% CI = 1.33-4.11). Maternal weight gain modified the effect of height on birth outcomes. Among women with inadequate weight gain, there was a 3-fold increase in risk for term-SGA (OR = 3.06; 95% CI = 2.41-3.89) for short maternal height, but not for preterm delivery. Among women with adequate plus weight gain, the association between short maternal height and term-SGA was 2.7 (95% CI = 0.55 9.43) compared to 3.67 (95% CI = 0.1.41-9.56) between short maternal height and preterm delivery. Maternal weight gain also modified the effect of maternal height on both outcomes for black mothers and for nonsmokers only.CONCLUSIONS: Findings indicate the need for interventions directed to those women who can benefit most from appropriate targeted weight gain recommendations. Therefore, prevention methods should be implemented to encourage short stature women to gain adequate weight during pregnancy. PMID- 11018416 TI - Methionine in the multifactorial etiology of neural tube defect affected pregnancies. AB - PURPOSE: Research in the last 40 years has provided evidence of the role of dietary nutrients in the development of NTDs. Failure of the neural tube to close is one of the most frequent and severe fetal developmental defects. The etiology of NTDs is complex encompassing genetic, dietary, and environmental factors. With regard to diet, it appears that in humans, a combination of low levels of methionine, folate, and Vitamin B(12) may lead to the occurrence of NTDs. The purpose of this study was to explore the relationship between maternal dietary intake of methionine and the risk of having a NTD affected pregnancy.METHODS: It was hypothesized that women with high maternal dietary methionine intake were at a decreased risk for NTDs. Combinations of methionine and folate without supplements, methionine and folate with supplements, and methionine and Vitamin B(12) and NTD risk were also examined. Data from a case-control study of 170 NTD affected pregnancies and 269 controls were provided by a CDC sponsored NTD Surveillance, Prevention, and Research Project.RESULTS: There was an approximately 30-55% reduction in NTD risk among women whose average daily dietary intake of methionine was greater than the lowest quartile of intake (>1580 mg/day). The odds ratios (95% CI) associated with the 3 quartiles of methionine intake greater than 1580 mg/day after adjusting for Kcal, race, and BMI were 0.72 (0.4030-1.288), 0.68 (0.3390-1.347), and 0.45 (0.1830 1.088).CONCLUSIONS: These findings indicate a reduction in the risk of having a NTD affected pregnancy is associated with maternal periconceptional dietary intake of methionine. PMID- 11018417 TI - Periconceptional vitamin use, dietary folate and occurrent neural tube defected pregnancies in a high risk population. AB - PURPOSE: This 5 year (1992-1997) population-based case control study was conducted to determine if supplemental or dietary folate taken during the periconceptional period reduces the risk of occurrent neural tube defect (NTD) affected pregnancies in a high risk population.METHODS: Cases were all NTD pregnancies including elective terminations, fetal deaths and live births in South Carolina. Only isolated, occurrent NTD affected pregnancies were included (N = 179). Controls (N = 288) were randomly selected from each hospital in proportion to the hospital's contribution to the total live births in the state for the previous year. All mothers were interviewed in person using a modified Centers for Disease Control and Prevention Birth Defect Risk Factor Surveillance Mother's Questionnaire and the Harvard Food Frequency Questionnaire.RESULTS: Women who used a vitamin with 0.4 mg or more of folate at least 3 times per week during the 3 months before and 3 months after conception had a lower risk of having an NTD affected pregnancy than women who took no folate supplementation at any time during pregnancy [adjusted odds ratio (AOR) = 0.42; 95% confidence interval (CI) = 0.19-0.94]; women who used a vitamin with 0.4 mg or more of folic acid for less than 3 times per week or in partial months during the 6 month periconceptional period also had a slightly lower risk of having an NTD affected pregnancy relative to women who used no folate supplementation at any time during pregnancy [AOR =.72; 95% CI = 0.41-1.30]. The risk of NTD affected pregnancy was also lower for women who daily consumed foods containing higher levels of folate: AORs were 0.36 (95% CI = 0.18-0.73), 0.72 (95% CI = 0.42-1.22), and 0.61 (95% CI 0.36-1.04) from the highest to lowest quartile of dietary folate which were 0.457 3.12 mg, 0.322-0.457 mg, and 0.236-0.322 mg respectively.CONCLUSIONS: A reduction in occurrent NTD risk associated with folate supplementation and dietary intake was observed. PMID- 11018418 TI - Prenatal tea consumption and risks of anencephaly and spina bifida. AB - PURPOSE: To evaluate the relationship between prenatal tea consumption and risk of anencephaly and spina bifida.METHODS: Data from the population-based Atlanta Birth Defects Case-Control Study were examined. Cases were infants with anencephaly (n = 122) or spina bifida (r = 154) and no other associated anomalies, and identified between 1968 and 1980. Controls were infants without birth defects (n = 3029) identified from birth certificates of the same birth cohort and frequency matched to cases by race, period of birth, and hospital of birth.RESULTS: Maternal tea consumption during the periconceptional period (3 months before through the first trimester of pregnancy) was reported at 82, 83.6, and 92.9% among controls, anencephaly, and spina bifida cases, respectively. With subjects whose mothers consumed no tea as a reference, odds ratios (OR) for tea consumption during the periconceptional period (adjusted for gender, race, period of birth, maternal age, education, alcohol consumption, smoking, and periconceptional multivitamins) were: anencephaly 0.9 (95% confidence limits (CI) 0.5-1.5); spina bifida 2.3 (CI 1.2-4.4). Odds ratios for spina bifida and number of cups of tea consumed/day were: 1-2 cups 2.1 (CI 1.1-4.0); 3+ cups 2.8 (CI 1.4 5.6). Consumption of other caffeinated beverages was not associated with risk for anencephaly or spina bifida.CONCLUSIONS: Further studies are warranted to corroborate and elucidate the observed association between tea consumption and spina bifida. PMID- 11018419 TI - Fatty acid composition of the red blood cell membrane in relation to menopausal status. AB - PURPOSE: Menopausal status effects female anatomical functioning at a variety of system-wide and cellular levels, including cellular membrane composition. This study analyzed a nested case-control ORDET data set of 433 pre and post menopausal breast cancer controls to examine the effects of menopausal status on the fatty acid composition of the red blood cell membrane.METHODS: ORDET is a prospective cohort study conducted in Italy to investigate the etiologic role of hormones and diet in breast cancer development. The fatty acid composition was measured and analyzed by gas chromotography, comparing retention time with standard measurement. Twenty-two individual fatty acids were measured, recorded, and categorized into four fatty acid groups: saturated, monounsaturated, polyunsaturated n-6 (PUFA n-6), and polyunsaturated (PUFA n-3) fatty acids.RESULTS: Post-menopausal women had consistently lower mean values for all four fatty acid categories and all individual fatty acids. Statistically significant mean differences, by menopausal status, were observed for three of the four fatty acid categories: saturated fatty acids (p = 0.006), PUFA n-6 acids (p = 0.001), and PUFA n-3 acids (p = 0.000). The biggest statistically significant differences in mean values among individual fatty acids for each category were observed for Palmitic acid (p = 0.009), Oleic acid (p = 0.040), Linoleic acid (p = 0.000), and Docosahexaenoic acid (p = 0.000). Individual fatty acids were also less highly correlated among post-menopausal women.CONCLUSIONS: There was an observed relationship between menopausal status and the fatty acid composition of the red blood cell membrane that warrants further study. This relationship may contribute to the physiological and psychological changes that occur during and after menopause, and may have far-reaching implications for women's health. PMID- 11018420 TI - Geographic information systems. A new tool in environmental epidemiology. AB - PURPOSE: Geographic Information Systems (GIS) are useful tools for identifying populations with potential exposure to environmental contaminants. Using a GIS, features of the local environment around an individual's home, work, or school can be described. We present two examples illustrating methods and issues in identifying populations potentially exposed to agricultural pesticides and to toxic releases from the Toxic Release Inventory (TRI).METHODS: We used USDA Farm Service Agency records as ground reference data to classify a late summer 1984 satellite image into crop species in 3 counties in Nebraska. We located residences from a case-control study of non-Hodgkin's lymphoma (NHL) on the crop maps and calculated the distance to crop fields. Residences from a 4-center study of NHL were mapped and the distance to TRI sites was determined.RESULTS: Twenty two percent of residences had crop fields within 500 meters of the home, an intermediate distance for the range of drift effects from pesticide applications. After accounting for the extent of primary drift from ground applications of pesticides, we estimated that 30 percent of residences were potentially exposed to crop pesticides. In the 4-center study, residence locations determined by address-matching methods and by a global positioning system were compared; the population 1 mile from specific TRI sites is described.CONCLUSIONS: These examples demonstrate the utility of a GIS in environmental epidemiology studies. A GIS can be a useful addition to questionnaire and other methods of exposure assessment in health studies. PMID- 11018421 TI - The effect of imputation of exposure estimates on the association between fine particulate matter and mortality. AB - PURPOSE: The Harvard Six Cities Study (HSCS) found a small but significant association between daily PM2.5 and daily mortality count. The HSCS findings have been used as the basis for new EPA regulations, requiring lower levels of PM2.5. We feel that there are unresolved issues regarding the HSCS that should be fully evaluated prior to its findings being used as the basis of new regulation, including how the extent and method of imputing exposure data affect the association with daily mortality counts.METHODS: We examined the association between PM2.5 levels and daily mortality count, comparing the results from the HSCS methods with results based on an alternate imputation method, and with non missing data.RESULTS: Overall, approximately 30% of the data points used in the HSCS were imputed. The method of imputation affected the association between particulate matter and mortality to a substantial degree in most of the cities. When the model using the HSCS method was compared to the model using the alternate method, in two areas the coefficients decreased substantially and lost significance. In two areas they changed little; in one area it rose substantially and became significant; and in one area it declined substantially but remained significant. When compared to the model based on the non-missing data, somewhat different patterns were observed. In both comparisons there were some large changes in the magnitude of the effect, but these were not consistent with the model used.CONCLUSIONS: We are concerned about the degree of data imputation and the effect that the method of imputation has on the association between particulate matter levels and mortality. In the case of the HSCS it appears that the imputed data are more strongly associated with the outcome than other methods of imputation and than the non-missing data. The reasons for these observations are not readily apparent, but the differences in effect should be explored and explained. PMID- 11018422 TI - Work-related fatalities in west virginia. A summary of surveillance, investigation, and prevention activities, july 1996-DECEMBER 1999. AB - PURPOSE: From 1990-95, West Virginia (WV) had a work-related death (WRD) rate of 8.9 deaths per 100,000-the fifth highest rate among all states and twice the national rate. As a result, a Fatality Assessment and Control Evaluation (WV FACE) program was established to identify all WRDs, define workers at high risk for fatal injury, investgate selected causes, and formulate and disseminate prevention strategies.METHODS: Surveillance and investigation data were used to describe trends and rates and identify hazardous conditions, unsafe work practices, and management-leadership problems through the use of the traditional epidemiologic model and the Haddon temporal matrix. Prevention strategies were developed and disseminated utilizing various written media.RESULTS: From 7/1996 12/1999, 191 persons died from work-related injuries. The WRD rate was 7.6 per 100,000 compared to 4.7 for the U.S. (1996-98). 94% of the victims were male and all Caucasian. Mean age at death was 43 years. Leading external causes of death: motor vehicle (48), struck by object (38), machinery-related (24), fall from elevation (15), and homicide (10). WRDs occurred most often in the trans/public utilities (37; truckers-22), manufacturing (32; loggers-24), mining (28), construction (26), and services (25) industry sectors. 19 on-site investigations were conducted (10 logging, 5 machine-related, and 4 fall from elevation); no company safety programs, inadequate training, lack of oversight were consistently noted. Summary reports were prepared for employers, Fatal Incident Alerts written for workers, and a scientific article was published.CONCLUSIONS: FACE has contributed to a better understanding of fatal occupational injuries within WV and the importance of coordinated efforts by employees, employers, and safety and public health specialists to reduce the frequency and societal impact of these injuries. PMID- 11018423 TI - Reliability of reporting on lifestyle and agricultural factors by a sample of participants in the agricultural health study from iowa. AB - PURPOSE: Information on agricultural practices has been obtained by questionnaire in several epidemiologic investigations. This project evaluated the reliability of self-reported information on pesticide use and various demographic and lifestyle factors among a group of farmers from Iowa.METHODS: 2,921 Iowa farmers participating in the Agricultural Health Study completed enrollment questionnaires approximately one year apart. Responses on the two questionnaires were compared for percent agreement and by Kappa statistics to evaluate reliability.RESULTS: Percent agreement for ever/never use of specific pesticides and application practices was quite high and generally ranged from 70% to over 90% and did not vary by age or educational level. Kappas were typically in the 0.50 to 0.60 range. Agreement was lower (typically 50% to 60%) for duration or frequency of use of specific pesticides. Agreement on lifestyle and non agricultural factors was comparable to that reported in other studies.CONCLUSIONS: Level of agreement regarding pesticide use in this population is similar to that found for diet, physical activity, and medical conditions, which have been successfully evaluated in many epidemiologic studies. Information on agricultural practices from self-completed questionnaires has sufficient reliability for use in epidemiologic investigations. PMID- 11018424 TI - Urinary biomarkers of atrazine exposure among farm pesticide applicators. AB - PURPOSE: This study assessed the feasibility of three laboratory methods for the detection of atrazine, a triazine herbicide, and its related metabolites in urine collected from field applicators.METHODS: Urine samples were collected from 256 randomly sampled field applicators 8 hours post application. Of these, 99 reported atrazine use during the application prior to sample collection and these samples were subsequently analyzed for urinary biomarkers.RESULTS: 37.4% (n = 37) samples showed detectable levels (minimum = 1.0 ng/mL) of deethylatrazine using gas chromatographic mass spectrometry (GCMS) analysis (X = 14.2 ng/mL; s.d. = 13.5). Fifty samples were tested using atrazine mercapturate in urine ELISA methods and 80% (n = 40) of these samples showed detectable levels of atrazine (X = 6.4 ng/mL; s.d. = 7.5). Of 10 samples tested by triazines in water ELISA methods, a common assay used for the detection of atrazine in groundwater, 100% showed detectable levels of atrazine (X = 22.4 ng/mL; s.d. = 13.9). Of the 21 samples collected from non-applicators and tested by GCMS, none evidenced detectable atrazine levels. Using GCMS as the gold standard, analyses showed that the mercapturate in urine ELISA was 48% sensitive and 91% specific whereas the triazines in water ELISA was 69% sensitive and 100% specific.CONCLUSIONS: It is possible to detect one-time atrazine exposures through analysis of urinary biomarkers among pesticide applicators. The feasibility of triazines in water ELISA methods for use in field studies for analyzing the presence of atrazine and related metabolites in urine was supported, but these methods need further testing on larger applicator samples before they can be used for standard screening. PMID- 11018425 TI - Magnetic field exposure of commercial airline pilots. AB - PURPOSE: Airline pilots are exposed to magnetic fields generated by the aircraft's electrical and electronic systems. The purpose of this study was to directly measure the flight deck magnetic fields to which commercial airline pilots are exposed when flying on different aircraft types over a 75-hour flight duty month.METHODS: Magentic field measurements were taken using personal dosimeters capable of measuring magnetic fields in the 40-800 Hz frequency range. Dosimeters were carried by either the Captain or the First Officer on Boeing 737/200, Boeing 747/400, Boeing 767/300ER, and Airbus 320 aircraft. The data were analyzed by aircraft type, with statistics based on block hours. Block hours begin when the aircraft departs the gate prior to take off and end when the aircraft returns to the gate after landing.RESULTS: Approximately 1008 block hours were recorded at a sampling rate of 3 seconds. Total block time exposure to the pilots ranged from a harmonic geometric mean of 6.7 milliGauss (mG) for the Boeing 767/300ER to 12.7 mG for the Boeing 737/200.CONCLUSIONS: Measured flight deck magnetic field levels were substantially above the 0.8 to 1 mG level typically found in the home or office and suggest the need for further study to evaluate potential health effects of long-term exposure. PMID- 11018426 TI - Nested case-control study of leukemia among a cohort of persons exposed to ionizing radiation from nuclear weapon tests in kazakhstan (1949-1963). AB - PURPOSE: A unique opportunity for epidemiological studies of cancer and other health effects of radiation exposures exists around the Semipalatinsk Nuclear Test Site in Kazakhstan. The present study is the first analysis of leukemia risk among the residents of downwind settlements exposed to radioactive fallout from atmospheric nuclear weapon tests (1949-1963) and followed up from 1960 to 1998.METHODS: Within the cohort of 10,000 exposed subjects a case-control study was nested, including 22 leukaemia cases (except chronic lymphoid leukemia) and 132 controls individually (1:6 ratio) matched by birth year and sex. Leukemia deaths were identified by death certificates and diagnoses were verified by hospital records. The individual dose including internal and external exposure assessment was estimated according to the residency and age at exposure. All odds ratios were adjusted for ethnicity (Russian or Kazakh) as an independent variable.RESULTS: The median dose of exposure for all subjects was 0.89 Sv ranging from 0.01 to 5.71 Sv. A nearly two-fold increased risk of leukemia was found (OR = 1.91; 95% CI = 0.38 to 9.67) for persons exposed to doses of >2.0 Sv as compared to those exposed to <0.5 Sv, but no increase in risk with the dose was found for those exposed to doses lower than 2 Sv. Detailed evaluation of dose response showed an excess relative risk for leukemia of 10% per 1 Sv of additional exposure.CONCLUSIONS: Our findings suggest that there is an increased risk of leukemia among those exposed to >2 SV as compared to those exposed to <0.5 Sv, but this could have been a chance finding due to the small number of cases and low statistical power. PMID- 11018427 TI - Mortality among radiologic technologists in the united states (1926-1997). 2(nd) Follow up. AB - PURPOSE: To evaluate risk for all-cause and cause-specific mortality in a large, primarily female (73%) cohort of radiologic technologists.METHODS: The study consists of 145,915 radiation technologists, certified in the American Registry of Radiologic Technologists (1926-1982) and followed through 1997. Causes of death were obtained from death certificates or, more recently, through NDI Plus. Standardized Mortality Ratios (SMR) were computed and tests of homogeneity were performed to detect differences in mortality among causes. Poisson models were used to estimate risks using an internal comparison group.RESULTS: Significantly low SMRs were observed for all causes (0.76), all cancers (0.82), and diseases of circulatory system (0.69). Compared to U.S. women, the risk for breast cancer mortality bordered around unity (SMR 1.01, 95% CI 0.94-1.09). However, relative to all other cancers, breast cancer mortality was significantly increased (RSMR 1.24, p < 0.01). Elevated risk for breast cancer was associated with certification before 1940 (SMR 1.55, 95% CI 1.24-1.91), and duration of certification of 20-29 (SMR 1.21, 95% CI 1.06-1.37) and 30+ years (SMR 1.77, 95% CI 1.54-2.02). A 35% increase in leukemia risk was evident for women certified for a duration of 20-29 years and a 36% increase among women certified for 30+ years. Poisson analysis revealed a significant increase in breast cancer risk with increasing number of years certified among women first certified before 1940 (p < 0.001) and during 1940-49 (p = 0.05) compared to women first certified in 1950 or later.CONCLUSIONS: Preliminary findings of this study suggest increased breast cancer risk associated with occupational radiation exposures prior to 1950 and with long-term cumulative exposures. However, potential confounding by reproductive and other risk factors needs to be evaluated. PMID- 11018428 TI - Enduring social and behavioral effects of exposure to military combat in vietnam. AB - PURPOSE: To assess current social and behavioral outcomes among Vietnam veterans up to 35 years after exposure to military combat.METHODS: A cohort of 1,315 American Legion members whose combat experiences were assessed with a standard quantitative measure in 1984 were followed-up through 1998 for current health and psychosocial status. Another 2,260 Legionnaires who served outside Vietnam during the same period were used as a comparison group. Outcomes included smoking and drinking habits as health risk indicators, income, measures of marital and general happiness and life satisfaction, and the Columbia Univerity PERI (Psychiatric Epidemiology Research Instrument) scales for psychological well being: anxiety, depression, helplessness/hopelessness, anger and irritation, and physical symptoms of depression. Combat intensity was classified as high, medium, and low.RESULTS: Income was significantly lower in the high-combat group by $4,000 per year compared to the low- and medium-combat groups and the non Southeast Asia veterans, whose incomes were similar to each other. Marital happiness, life happiness, general life satisfaction, decreased with increasing combat, while smoking prevalence increased with combat score. The PERI scale components were strongly dose-related to combat intensity. Furthermore, the anger/irritation score for the low-combat group was significantly higher than for the non-SEA group.CONCLUSIONS: Although practically all veterans in this study last saw combat prior to 1971, the detrimental effects of having experienced combat which were strongly apparent when this group was surveyed in the 1980s continue to exert an adverse effect on many aspects of their lives almost three decades after the war. PMID- 11018429 TI - A nested case-control study of stomach cancer mortality among automobile machinists exposed to metalworking fluid. AB - PURPOSE: In a nested case-control study of 140 stomach cancer deaths (1941-1984) among 46,384 automobile workers, we examined risk in relation to lifetime exposure to metalworking fluid (MWF) in machining and grinding operations.METHODS: Exposure was defined during varying windows of biologic time (<10 years before death, 10-20 years before death, and > 20 years before death) and evaluated using a nonparametric smoothing technique and conditional logistic regression. Exposure was measured by duration, intensity (mg/m(3)), and cumulative exposure (mg/m(3)-years) to each MWF type (straight, soluble, or synthetic), as well as by duration of exposure to selected metals, additives, and components of the fluids.RESULTS: When data from three plants were combined, grinding with water-based synthetic or soluble MWF increased the risk of stomach cancer mortality. Those grinding with synthetic MWF with more than 1.3 mg/m(3) years exposure during the 10 years just prior to death experienced a 4.4-fold risk of stomach cancer (95% CI 1.5-13.1). Increased risk was also observed among those in the highest category of soluble grinding grading exposure (OR 1.9; 85% CI 1.0-3.6). In plant-specific analysis, there was evidence of increased risk of stomach cancer associated with exposure to straight MWF and iron, as well as the water-based fluids.CONCLUSIONS: Study findings suggest that stomach cancer mortality was associated with exposure to each MWF type. Results suggesting that synthetic MWF may act as a stomach cancer promoter are of particular concern because this MWF is widely used in today's high speed grinding operations. PMID- 11018430 TI - Job exposures as correlates of recovery in population-based rehabilitation intervention for work-related low back disorders. AB - PURPOSE: Evaluating the impact of population-based rehabilitation interventions for work-related low back disorders (WR-LBD) requires job exposure factors to be considered as time-varying covariates. The role of job factors in recovery has not been well-established as most studies are based upon clinic samples, not working populations. This report represents the initial exploration of variables to be included in modeling job exposures as time-varying covariates of a study of work-place based rehabilitation intervention.METHODS: The study sample consisted of 162 hourly production employees from two automotive plants with work-related low back disorder not due to external trauma. Data reported herein were collected at baseline from an on-going large randomized clinical trial of rehabilitation for WR-LBD. Low back pain as measured by the North American Spine Society Baseline Form was examined in relation to: self-reported job factors from the Job Content Questionnaire and job risk category for low back disorder determined by the Lumbar Motion Monitor (LMM), an electronic goniometer which transmits back motion signals to a computer for analysis.RESULTS: Low back pain, with and without leg pain, was not found to be associated with hours worked in the previous week, job shift, perceptions of the pace of the job, or job risk of low back disorder as measured by the LMM. Low back pain was significantly (p < 0.05) correlated with perceptions of: how hard the work was, how much physical effort was required, how hectic the job was, whether or not there was sufficient time to do the job, and how much concentration and physical effort were required for the job.CONCLUSIONS: Self-report of perceptions of job strain may be as important, if not more important, than current quantitative factors thought to influence recovery from work-related low back pain. PMID- 11018431 TI - Gender differences in carpal tunnel syndrome? occupational and non-occupational risk factors in a population-based case-control study. AB - PURPOSE: To determine the role of occupational and personal risk factors of carpal tunnel syndrome (CTS) separately in men and women.METHODS: Men (n = 404) and women (n = 404) younger than 65 years with first surgery for CTS in 1995/96 were sampled at random from the Bremen population (0.65 million) and matched to controls by age and gender sampled from the population registry. A self administered questionnaire with 77 questions inquired about personal factors and activities during work and private environment (response: 60% of eligible cases and 52% in controls, respectively).RESULTS: CTS risk increased with body mass index (BMI): odds ratio for each unit of BMI [kg/m(2)] OR(men) = 1.13; (95% confidence-limits CL(men) 1.06, 1.20), OR(women) = 1.09 (CL(women) 1.04, 1.14). Multivariate analysis adjusting for BMI showed more pronounced risks in men compared with women for repetitive movements of the hand: OR(m) = 2.89 (CL 1.82, 4.58); OR(w) = 2.10 (CL 1.37, 3.22), for forceful grip: OR(m) = 2.69 (CL 1.70, 4.27); OR(w) = 2.29 (CL 1.43, 3.66), but not for household chores: OR(m) = 0.64 (CL 0.40, 1.03); OR(w) = 0.88 (CL 0.34, 2.29). Women worked less hours per week than men but manual exposures remained more relevant in men after adjustment. Exposure-response-relationships could be demonstrated for all work related exposures. Women were at higher risk for CTS if they had had more than two births or a history of hysterectomy. Adding these factors to the logistic model still showed an association between manual work and CTS. Blinded expert rating (manual vs. non-manual) of jobs held by respondents and non-respondents and by cases and controls did not reveal relevant selection effects or recall-bias.CONCLUSIONS: CTS is a work related disease in both men and women, the fraction attributable to work in the Bremen population under age 65 is estimated to be 33% in men and 15% in women. Funding: German Federal Ministry for Education & Research: 01 EG 9512. PMID- 11018432 TI - Correlates of the "western" and "prudent" diet patterns in the us. AB - PURPOSE: Underlying the diversity and changeability of US eating habits are fundamental dietary patterns shaped by common sociohistorical experiences. Currently, little is known of what patterns exist or their associations with sociodemographic/lifestyle factors. We examined these issues in third National Health and Nutrition Examination Survey data.METHODS: Analyses were based on 5,794 US-born, white participants. Interviews included a 60-item food frequency questionnaire (FFQ) and questions on sociodemographics and health behaviors. Dietary patterns were identified by principal components analysis on FFQ responses. Component scores representing intake level of each pattern were dichotomized at the median. Logistic regression analysis was used to estimate odds ratios (OR) and 95% confidence intervals (CI).RESULTS: We identified two primary dietary patterns-a "western" pattern characterized by processed and red meats, eggs, potatoes, and refined grains, and a "prudent" pattern characterized by cruciferous vegetables, greens, carrots, salads, and fresh fruits. The two patterns occurred across geographic regions and in both women and men. After age- and sex-adjustment, the "western" diet was associated with rural residence (OR = 1.9, 95% CI 1.7-2.2), working class status (OR = 1.4, 95% CI 1.2-1.6), and lack of high school completion (OR = 1.6, 95% CI 1.4-1.8). The "prudent" diet was associated with high school completion (OR = 2.8, 95% CI 2.5-3.3) and behaviors reflecting greater health awareness, such as daily physical activity (OR = 2.0, 95% CI 1.8-2.2), non-smoking (OR = 2.1, 95% CI 1.8-2.4), and supplement use (OR = 1.8, 95% CI 1.6-2.0).CONCLUSIONS: The two dietary patterns resemble those found in previous US studies. Findings regarding correlates agree with social histories describing the emergence of those patterns. Principal components analysis is potentially useful for identifying fundamental dietary patterns for future investigations of diet-disease associations. PMID- 11018433 TI - Differences in alcohol risk profiles between college students and college-age non students presenting for care in the emergency department. AB - PURPOSE: Determine whether college students and non-students seeking emergency medical care experience different lifestyle choices, alcohol-use patterns, and risk profiles.METHODS: Patients aged 18-25 seen in a university-affiliated ED between August 1998 and June 1999 who had >/=1 drinks within the past year were eligible. Patients scoring >5 out of a possible 40 on the Alcohol Use Disorder Identification Test (AUDIT) were considered at elevated risk for alcohol problems and were interviewed further to determine alcohol-use patterns and history.RESULTS: Of 1,436 consenting patients, students were more likely to screen positive for alcohol problems than non-students (55% vs 43%; 478/870 vs 245/566). Furthermore, among screen-positive patients, students were more likely than non-students to binge drink in the two weeks prior to screening (82% vs 65%) and to drink illegally (60% vs 38%). Students needed fewer drinks to feel the effects of alcohol (3.9 drinks vs 5.1) and were less likely to request a referral for alcohol treatment (2% vs 8%). However, students had lower baseline risk for alcohol problems than non-students (AUDIT = 11.2 vs 13.2). Students were younger than non-students (20.2 years vs 21.4), older at their first drunken experience (15.3 vs 14.7), and less likely to smoke (51% vs 76%) or have a family history of heavy drinking (5% vs 23%). (p <.01 for all comparisons).CONCLUSIONS: Students exhibit a higher risk than non-students for behaviors associated with acute alcohol-related problems. However, they may be at lower risk for chronic alcohol problems. This university-based ED seems an appropriate venue for early identification and brief, on-site intervention for students with alcohol problems. PMID- 11018434 TI - Gun availability, psychopathology, and risk of death from suicide attempt by gun. AB - PURPOSE: Suicide is the third leading cause of death among adolescents and the 8th leading cause of death overall. Guns figure prominently in this problem. Guns are the most lethal means of suicide, and in the United States about 60% of all suicides are committed by guns. We examined hospital admissions for suicide attempts (para-suicide) in the city of Chicago between 1990-1997. Our aim was to 1) quantify the risk of death from para-suicide by gun versus other methods, 2) determine whether individual with psychopathology are more likely than others to attempt suicide by gun, 3) determine proportion of variation in para-suicide by gun explained by community-level gun availability independent of individual-level determinants of para-suicide by gun.METHODS: Hospital admissions from city of Chicago during 1990-1997 were examined for cases of attempted or completed suicide. 11,584 individuals were identified with a diagnosis of para-suicide (E950-E959, classifiable to ICD-9CM). These individuals were then classified into suicide type (e.g., gun, crash, cut, poison) and were also identified as having a mental disorder or not (mental diagnoses classifiable to ICD-9CM).RESULTS: 1) Those attempting suicide by gun are about 70 times more likely to die from their injuries than those attempting suicide by other means. 2) Depressed and psychotic individuals compared with individuals with no-mental disorders are significantly more likely to attempt suicide with a gun (p < 0.001). 3) Inclusion of community level gun availability in a hierarchical model reduced the variation in the model by 16%.CONCLUSIONS: Guns are the most lethal suicide method. Community-level availability, independent of individual-level factors, is an important determinant of suicide attempt by gun. PMID- 11018435 TI - Screening for learning disabilities in preadolescents using psychosocial disability markers-a pilot study. AB - PURPOSE: Learning disabilities (LD) are among the most common developmental and psychiatric disorders encountered by pediatricians, yet many children are not adequately diagnosed or treated in elementary school when intervention is most effective. Because of the well-known co-morbidity between psychosocial problems and LD, this pilot study sought to utilize psychosocial "markers" to screen for unidentified LD among all fifth graders in three urban/suburb Connecticut elementary schools.METHODS: 115 fifth graders, representing a 71% response rate, agreed to participate in the study and were screened using standardized psychosocial questionnaires administered to parents (Child Behavior Checklist), students (Youth Self Report), and teachers (Teacher Report Form). Psychosocial symptom scores and associated t-scores were obtained for three types of problems; "internalizing," "attention" and "externalizing." 28 students with high levels (t > 60) of internalizing or attention problems, were selected along with a sample of 22 "normal" students with no evidence of psychosocial impairment. These 50 students were given an individually administered battery of ability, performance and cognitive processing tests to determine the presence of unidentified LD. Performance tests included standardized tests of reading, writing and mathematics. All test scores were normed with a mean of 100 and a SD of 15. LD was determined to be present if the student had a 1.5 SD or greater discrepancy between ability and performance in any of the three academic areas, and evidence of a cognitive processing deficit.RESULTS: The results indicated that 14 of the 50 students tested (28%) had evidence of undiagnosed LD. No significant difference was found in the proportion of undiagnosed LD among psychosocially impaired versus normal students. However regression analysis suggested that this finding was misleading. Controlling for ability, attention problems reported by the parent, were found to be significant (p < 0.05) predictors of lowered test scores, whereas internalizing problems were found to be predictive of increased test scores. Students with both internalizing and attention problems did not differ significantly from psychologically unimpaired students.CONCLUSIONS: This pilot study indicates that better screening methods are needed to identify the likely high proportion of elementary school children with undiagnosed learning difficulties and co-morbid psychosocial problems. PMID- 11018436 TI - The longitudinal study of phobic disorder in a community sample from early to late adolescence. AB - PURPOSE: This study explored the development of phobic disorder as young subjects move into late adolescence. The objectives were to determine: the frequency of clinical, subsyndromal and subthreshold phobia in late adolescence; the incident episode and stability of phobic symptoms as adolescents move from early to late adolescence; and the relationship between demographic characteristics in early adolescence and the presence of clinical, subsyndromal and subthreshold phobia in late adolescents.METHODS: The data were taken from a two-stage epidemiological study to determine the frequency of and risk factors for DSM-III major depressive disorders and suicidal behaviors in a community population of adolescents. Data were also collected for the frequency of phobias. Three levels of phobias were defined.RESULTS: The prevalences of clinical, subsyndromal and subthreshold phobias in late adolescence were 0.40%, 1.34%, and 5.74% respectively. The weighted incidence of clinical phobia was 0.28%, of subsyndromal phobia was 0.19% and of subthreshold phobia was 4.63%. Race, sex, and having a subthreshold phobia at the initial interview were significantly associated with subsyndromal phobia in late adolescence. Diagnosis of clinical phobia at initial interview was significant in all the multivariate models. Having a subthreshold phobia at initial interview was significant for subsyndromal phobia in late adolescence.CONCLUSIONS: Data on late adolescence showed a strong trend towards remission of phobic symptoms and disorder. However, those with clinical phobia at baseline had an increased risk of being diagnosed again at late adolescence. The most frequently reported symptoms were simple phobias, suggesting that simple phobias start early, have a higher prevalence, and are more frequent over time than social or agoraphobia. Further longitudinal studies are needed. PMID- 11018437 TI - Introduction to the special issue on the SENIAM European Concerted Action. PMID- 11018438 TI - Recent progress in the diagnostic use of surface EMG for neurological diseases. AB - The different techniques to measure and analyze surface EMG are summarized with an emphasis on the clinician's point of view. The application of surface EMG in neurological disease is hampered by many inherent problems, especially the difficulties in extracting features of single motor units. However, the evolution of surface EMG from single bipolar recordings via a linear array of multiple electrodes to densely packed, multi-channel electrode arrays could in principle solve this problem. The added value of using multiple channels (up to 128) with an interelectrode distance of a few millimetres to obtain more spatial information is emphasized. At least for some muscles it is now possible to extract information from the surface EMG, conventionally thought to belong to the domain of needle EMG (for example the "electrical size" of motor units). The use of analysis techniques such as the estimation of muscle fiber conduction velocity has already proven to be of diagnostic value in several myopathies characterized by a disturbed membrane function and in metabolic myopathies with abnormal fatigue profiles. Future research should be directed at the development of analysis techniques enabling the extraction of more relevant motor unit variables from surface EMG signals. PMID- 11018439 TI - Force and EMG power spectrum during and after eccentric and concentric fatigue. AB - Eccentric and concentric force and median frequency of the EMG power spectrum were measured during and immediately after maximal eccentric (EE) and concentric (CE) exercise and during the recovery period of 1 week. Eight male subjects performed EE and CE consisting of 100 maximal eccentric and concentric actions with elbow flexors during two separate exercise sessions. When comparing maximal eccentric and concentric actions before the exercises, the average force was higher (P<0.001) in eccentric than in concentric but the average rectified EMG (aEMG) values were the same with the two types of action. The average eccentric force decreased 53.3% after EE and 30.6% after CE, while the average concentric force decreased 49.9% after CE and 38.4% after EE. The recovery was slower after EE. The median frequency (MF) of biceps brachii (BB) in eccentric action decreased during both EE (P<0.01) and CE (P<0.05). It recovered within 2 days of the exercises but was lower again (P<0.01) 7 days after EE. In concentric action MF of BB decreased during CE (P<0.01), while no changes were observed in EE. Blood lactate concentration increased (P<0.001) in both exercises and serum creatine kinase (CK) activity increased in EE only, being significantly higher (P<0.001) 7 days after than before the eccentric exercise. In the absolute scale, the eccentric force in EE decreased more than the concentric force in CE (P<0.01). Fatigue response was action type specific as seen in the greater reduction in the force of the exercise type. MF decreased immediately after both exercises, which may be at least partly related to elevated blood lactate concentration. Eccentric actions led to possible muscle damage as indicated by elevated serum CK and muscle soreness, and therefore to longer recovery as compared to concentric actions. Decreased MF after EE may be indicative of selective damage of the fast twitch fibers in this type of exercise. PMID- 11018440 TI - Methodologies for evaluating electromyographic field data in ergonomics. AB - Surface electromyography (SEMG) is an important tool for work load assessment in ergonomics. Several different approaches using amplitude as well as frequency parameters give fruitful information depending on question at issue in the laboratory as well as in field studies. One basic factor determining the choice of analysis method is whether the SEMG is interpreted as an indicator of forces/torques or pure muscular activation. Two methods for occupational SEMG data reduction representing two different approaches to SEMG applications in ergonomics, Exposure Variation Analysis (EVA), and Joint Analysis of EMG Spectrum and Amplitude (JASA), applied on the same SEMG recording from three muscles during urology surgeon work, have been compared. The EVA method categorised the three muscle recordings as too static with no EMG gaps while the JASA method identifies fatigue in two of the three recordings. The practical relevance of these findings is discussed. PMID- 11018441 TI - Surface EMG models: properties and applications. AB - After a general introduction on the kind of models and the use of models in the natural sciences, the main body of this paper reviews potential properties of structure based surface EMG (sEMG) models. The specific peculiarities of the categories (i) source description, (ii) motor unit structure, (iii) volume conduction, (iv) recording configurations and (v) recruitment and firing behaviour are discussed. For a specific goal, not all aspects conceivable have to be part of a model description. Therefore, finally an attempt is made to integrate the 'question level' and the 'model property level' in a matrix providing direction to the development and application of sEMG models with different characteristics and varying complexity. From this overview it appears that the least complex are models describing how the morphological muscle features are reflected in multi-channel EMG measurements. The most challenging questions in terms of model complexity are related to supporting the diagnosis of neuromuscular disorders. PMID- 11018442 TI - Geometrical factors in surface EMG of the vastus medialis and lateralis muscles. AB - Surface EMG signals detected in dynamic conditions are affected by a number of artefacts. Among them geometrical factors play an important role. During movement the muscle slides with respect to the skin because of the variation of its length. Such a shift can considerably modify sEMG amplitude. The purpose of this work is to assess geometrical artefacts on sEMG during isometric contractions at different muscle lengths. The average rectified value (ARV) of 15 single differential signals was obtained by means of a linear array of 16 bar electrodes from the vastus medialis and lateralis muscles. The knee angle was changed from 75 degrees to 165 degrees in steps of 30 degrees and voluntary isometric contractions at a low, medium and high force level were performed for each angle. The ARV pattern was normalized with respect to the mean activity to compare signals from different joint angles. From the data collected it was possible to separate the geometrical changes from the changes due to different intensities of activation. In three out of five subjects, we found (within the resolution of our measures) a 1 cm shift for the vastus medialis muscle while no shift was observed for the other two subjects. For the vastus lateralis muscle a 1 cm shift was found in two out of four subjects. Such a shift produces the main contribution to geometrical artefacts. To avoid such artefacts the innervation zones should be located and the EMG electrodes should not be placed near them. PMID- 11018443 TI - Comparison of algorithms for estimation of EMG variables during voluntary isometric contractions. AB - Many algorithms have been described in the literature for estimating amplitude, frequency variables and conduction velocity of the surface EMG signal detected during voluntary contractions. They have been used in different application areas for the non invasive assessment of muscle functions. Although many studies have focused on the comparison of different methods for information extraction from surface EMG signals, they have been carried out under different conditions and a complete comparison is not available. It is the purpose of this paper to briefly review the most frequently used algorithms for EMG variable estimation, compare them using computer generated as well as real signals and outline the advantages and drawbacks of each. In particular the paper focuses on the issue of EMG amplitude estimation with and without pre-whitening of the signal, mean and median frequency estimation with periodogram and autoregressive based algorithms both in stationary and non-stationary conditions, delay estimation for the calculation of muscle fiber conduction velocity. PMID- 11018444 TI - EMG signals detection and processing for on-line control of functional electrical stimulation. AB - The surface EMG signal detected from voluntarily activated muscles can be used as a control signal for functional neuromuscular electrical stimulation. A proper positioning of the recording electrodes in relation to the stimulation electrodes, and a proper processing of the recorded signals is required to reduce the stimulus artefact and the non-voluntary contribution (M-wave). Six orientations and six locations of the recording electrodes were investigated in the present work. A comb filter (with and without a blanking windowing) was applied to remove the signal components synchronously correlated to the stimulus. An operative definition of the signal to noise ratio and an efficiency index were implemented. It resulted that when the recording electrodes were located within the two stimulation electrodes the best orientation was perpendicular to the longitudinal line. However the best absolute indexes were obtained when the recording electrodes were located externally of the stimulation electrodes, and in that case the best orientation was longitudinal. Concerning the filtering procedure, the use of a blanking window before the application of the comb filter, gave the best performance. PMID- 11018445 TI - Development of recommendations for SEMG sensors and sensor placement procedures. AB - The knowledge of surface electromyography (SEMG) and the number of applications have increased considerably during the past ten years. However, most methodological developments have taken place locally, resulting in different methodologies among the different groups of users.A specific objective of the European concerted action SENIAM (surface EMG for a non-invasive assessment of muscles) was, besides creating more collaboration among the various European groups, to develop recommendations on sensors, sensor placement, signal processing and modeling. This paper will present the process and the results of the development of the recommendations for the SEMG sensors and sensor placement procedures. Execution of the SENIAM sensor tasks, in the period 1996-1999, has been handled in a number of partly parallel and partly sequential activities. A literature scan was carried out on the use of sensors and sensor placement procedures in European laboratories. In total, 144 peer-reviewed papers were scanned on the applied SEMG sensor properties and sensor placement procedures. This showed a large variability of methodology as well as a rather insufficient description. A special workshop provided an overview on the scientific and clinical knowledge of the effects of sensor properties and sensor placement procedures on the SEMG characteristics. Based on the inventory, the results of the topical workshop and generally accepted state-of-the-art knowledge, a first proposal for sensors and sensor placement procedures was defined. Besides containing a general procedure and recommendations for sensor placement, this was worked out in detail for 27 different muscles. This proposal was evaluated in several European laboratories with respect to technical and practical aspects and also sent to all members of the SENIAM club (>100 members) together with a questionnaire to obtain their comments. Based on this evaluation the final recommendations of SENIAM were made and published (SENIAM 8: European recommendations for surface electromyography, 1999), both as a booklet and as a CD-ROM. In this way a common body of knowledge has been created on SEMG sensors and sensor placement properties as well as practical guidelines for the proper use of SEMG. PMID- 11018446 TI - Pneumocystis carinii enhances soluble mannose receptor production by macrophages. AB - Phagocytosis of extracellular organisms in the alveolar spaces of the lungs represents the first-line of host defense against pulmonary pathogens. Disruption of this process is likely to interfere with the generation of appropriate specific immune responses, and lead to a delayed or inefficient clearance of the pathogen. Pneumocystis carinii, an opportunistic pathogen in immunodeficient individuals, is cleared from the lung by alveolar macrophages. In the absence of specific anti-Pneumocystis antibodies, phagocytosis is dependent on the non opsonic macrophage mannose receptor (MR). Recent studies have demonstrated that alveolar macrophage MR activity is downregulated in individuals infected with HIV, and that functional MR is shed from the macrophage cell surface. Here we report that P. carinii enhances the formation of soluble MR by macrophages in vitro. Soluble MR was detected in cell-free alveolar fluid from humans infected with HIV and/or P. carinii, but not in alveolar fluid from healthy controls. Soluble MR was found in association with extracellular clumps of P. carinii in the lungs of mice with P. carinii pneumonia, and was associated with P. carinii organisms purified from these mice. When purified P. carinii organisms were incubated with soluble MR-containing supernatants, they were phagocytosed less readily by alveolar macrophages than were control organisms. Our results suggest that P. carinii organisms enhance the shedding of MR from the surface of alveolar macrophages, and that the resultant soluble MR binds to intra-alveolar organisms, thereby interfering with their non-opsonic uptake via the macrophage cell surface MR. PMID- 11018447 TI - Characterisation of NF-kappa B complexes in Theileria parva-transformedT cells. AB - Transformation of T cells by the intracellular parasite Theileria parva is accompanied by constitutive I-kappa B degradation and NF-kappa B activation, a process which is essential to prevent the spontaneous apoptosis of these parasite transformed cells. NF-kappa B-mediated responses are regulated by selective combinations of NF-kappa B proteins as homo- or heterodimers and by distinct kappa B motifs. We characterised the NF-kappa B complexes induced by T. parva infection in TpM(803) T cells. By western blot, we demonstrated that all members of the NF-kappa B/Rel family of proteins translocate to the nucleus of infected cells. Using two different kappa B oligonucleotides (kappa B-1 and kappa B-2), both containing the decameric consensus kappa B motif (GGGACTTTCC), clearly distinct patterns of DNA binding activities could be demonstrated in electrophoretic mobility shift assays. Supershift analysis and UV cross-linking assays showed that complexes binding to kappa B-1 consisted of p50, p65 and RelB homo and/or heterodimers. We could also detect an association of ATF-2 and c-Fos with one of the complexes. The HIV-derived kappa B-2 oligo only bound p50 and p65. Additionally, several agents known to inhibit a wide range of NF-kappa B activation pathways had no inhibitory effect on the activation of NF-kappa B DNA binding in TpM(803) T cells. PMID- 11018448 TI - Replication and virulence of early protein 0 and long latency transcript deficient mutants of the Aujeszky's disease (pseudorabies) virus. AB - Early protein 0 (EP0)-deficient recombinant Aujeszky's disease viruses, Ka-ep0lac and Ba-ep0lac derived from strains Kaplan and Bartha, respectively, were constructed to explore the impact of the mutation on replication, virulence and latency of the virus. Inactivation of the EP0 gene resulted in a mutation of long latency transcript (Cheung et al., 1991) that is located on the complementary DNA strand of EP0 and immediate early protein (IE)175 genes. In infection of immortalized porcine kidney cells, the growth rate and yield of both EP0(-) mutant strains were significantly smaller than that of wild-type virus. Ka-ep0lac was found to be highly virulent, while Ba-ep0lac showed an attenuated phenotype in mice. PCR assay and immunohistochemistry showed that the Ba-ep0lac virus was able to establish latency in the mouse trigeminal ganglia. However, latent virus was not able to reactivate in explant reactivation assays. Accordingly, latent Ba ep0lac has the potential to be exploited as vectors for the delivery of foreign genes to the nervous system. PMID- 11018449 TI - Apoptotic neurodegeneration induced by influenza A virus infection in the mouse brain. AB - Neurodegeneration in the brain induced by the WSN strain of influenza A virus was investigated after stereotaxic introduction into the olfactory bulb of C57BL/6 mice. Immunohistochemistry detected WSN virus-infected neurons in the anterior olfactory nucleus as early as day 3 postinfection. Thereafter, they became shrunken and showed loss of neurite-immunolabeling and chromatin condensation. Infected neurons died by day 12, degenerating into multiple small granular bodies. The terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling method demonstrated DNA fragmentation in infected neurons at day 7 and also in such granular bodies at day 12. In perforin-deficient mice, the appearance of virally induced apoptotic neurodegeneration was delayed and virus infection continued for a longer period of 35 days postinfection. These findings indicate that perforin-mediated neuroapoptosis appears significant in exterminating the intracellular pathogen at an early stage of infection. PMID- 11018450 TI - Do infectious diseases drive MHC diversity? AB - The primary function of the major histocompatibility complex (MHC) is to allow the immune system to identify infectious pathogens and eliminate them. Infectious diseases are now thought to be the main selection force that drives and maintains the extraordinary diversity of the MHC. PMID- 11018451 TI - Structure of the intestinal flora responsible for development of the gut immune system in a rodent model. AB - The intestinal flora comprising indigenous, autochthonous bacteria is constantly present in the alimentary tract of host animals, including humans. The indigenous bacteria greatly affect the structure and functions of the intestinal mucosa. Studies involving gnotobiotic mice or rats have shown that the presence of limited kinds of intestinal bacteria is responsible for the development of the gut immune system, such as secretory IgA, major histocompatibility complex molecules and intraepithelial lymphocytes. Understanding of the structure of the intestinal flora or the organization of the microbial population in the intestine, based on evaluation of the immunological responses, may clarify its functions in the host animal. PMID- 11018452 TI - Issues in tick vaccine development: identification and characterization of potential candidate vaccine antigens. AB - It is well established that acquired immunity against tick infestation can be induced by repeated tick infestation or by active immunization with either crude or purified native as well as recombinant antigens. This review provides insights into the development of tick vaccines with reference to identification, purification and molecular cloning of candidate target antigens. PMID- 11018454 TI - Nocardia brasiliensis: from microbe to human and experimental infections. AB - Nocardia brasiliensis is a Gram-positive bacterium that lives as a saprophyte in soil. In this article the physical properties, chemical composition and taxonomic position of this species is reviewed. Human infections and an experimental model of actinomycetoma in BALB/c mice as well as the host-immune response is described. PMID- 11018453 TI - The third-stage larva (L3) of Brugia: its role in immune modulation and protective immunity. AB - In this review, we focus on the role of the L3 (third-stage larva) of lymphatic filarial nematodes in immunomodulation and in the development of protective immunity. Studies in the mouse models of Brugia have been fundamental to our understanding of the mechanisms by which infection with L3 results in Th2 responses and the active suppression of Th1 responses. The relevance of these phenomena to the human infection is discussed. PMID- 11018455 TI - Immunotherapy against antibiotic-resistant bacteria: the Russian experience with an antistaphylococcal hyperimmune plasmaand immunoglobulin. AB - The Russian experience with the preparation and clinical application of an antitoxic antistaphylococcal hyperimmune plasma and immunoglobulin is described. The immunotherapies were developed in the late 1960s and put into widespread use in the Soviet Union for the prophylaxis and treatment of sepsis, pneumonia, and other conditions caused by an epidemic of antibiotic-resistant Staphylococcus aureus. PMID- 11018456 TI - Effects of loading conditions and objective function on three-dimensional shape optimization of femoral components of hip endoprostheses. AB - A numerical procedure was implemented for the three-dimensional (3D) shape optimization of the femoral component in total hip replacement. An algorithm was developed for defining the component geometry in terms of longitudinal and cross sectional shape variables. The 3D design model was combined with a 3D finite element analysis and a numerical optimization procedure. An idealized femoral geometry and perfectly bonded interfaces were used for cemented and uncemented implants. The design objective was to smooth some measure of the stresses along the local interface. The effects of two different load conditions and several different objective functions were examined. A common initial implant design was used for all cases. The general trend in all design optimization was to produce a somewhat bulky implant with a rectangular cross-section. The outcome was more strongly affected by loading condition than the choice of objective function. The use of a strain energy density criterion as the objective function proved to be the most effective in reducing all equivalent stress criteria. PMID- 11018457 TI - Theoretical study of convergent ultrasound hyperthermia for treating bone tumors. AB - This study investigates the optimal external parameters for using an ultrasound applicator for treating bone tumors. This system utilized spherically arranged applicators such as scanned focused ultrasound, and spherically focused multielement applicators. The power deposition pattern is modeled as geometric gain with exponential attenuation. The specific absorption rate ratio (SARR) criteria have been used to determine the proper heating domain of ultrasound driving frequency and therapeutic tumor diameter. The results demonstrate that the optimal driving frequency depends on tumor depth, ultrasound absorption of bone marrow, and diameter of bone, but it is independent of the acoustic window area and SARR. The treatable diameter of bone tumor increased when the absorption ratio of bone marrow to tumor, acoustic window of surface skin, and diameter of bone were elevated. However, the treatable diameter of bone tumor decreased when muscle thickness, SARR of bone tumor site to the surface skin, bone marrow, and bone declined. To deliver the ultrasound energy into the tumor site and to avoid the potential damage to the normal tissue as much as possible, the specific absorption rate (SAR) in the bone tumor site has to be three times higher than that in the surface skin, tumor/marrow, and marrow/bone interfaces. The temperature distributions can verify the SARR criteria in this model. This study provides the information for choosing the optimal operating frequency of the ultrasound transducer and the acoustic window on the skin surface, and for designing the ultrasound applicator for clinical implementation. PMID- 11018458 TI - Unsteady stenosis flow prediction: a comparative study of non-Newtonian models with operator splitting scheme. AB - This paper presents a comparative study of non-Newtonian and Newtonian models of blood. A non-Newtonian incompressible 2-D Navier-Stokes (N-S) solver has been developed using Fasttalk language within the Fastflo environment. It is based on the method of operator splitting with artificial compressibility technique. The Power law and Casson models have been used as the constitutive equations for blood with a hematocrit of approximately 45%. These two non-Newtonian models and the Newtonian model are used to simulate unsteady flow through a hypothetical stenotic geometry over an aperiodic time interval of 1 s. Through comparison of the results of the three models, it was found that the wall shear stress (WSS) distribution over the time interval was comparable for both non-Newtonian models. The peak WSS for the Newtonian model had the lowest value. The peak wall shear stress gradient (WSSG) for the Power law was the highest, followed by the Casson and Newtonian models. Flow characteristics such as higher pressure drop across the stenosis, location and movement of vortex were similar in all three models. Non-Newtonian effects were most significant in the vicinity of the stenosis. PMID- 11018459 TI - In vitro monitoring of rabbit anterior cruciate ligament damage by acoustic emission. AB - Anterior cruciate ligament (ACL) rupture is a major clinical problem leading to instability and degeneration of the knee joint. The problem is compounded by the limited ability of the ACL to heal when ruptured. The existing knowledge regarding the way the ACL ruptures is limited, and this investigation is an attempt to understand the nature of the ruptures using the rabbit as a model. A total of 16 rabbit tibia-ACL-femur complexes were stretched in tension to complete rupture. Four specimens were stretched to failure at a displacement rate of 0.5 mm/min and 12 specimens at 10 mm/min. Acoustic emission (AE) transducers were placed on both the tibia and the femur, and stress wave signals generated during the tensile test were recorded. Fibre fractures produced the highest amplitude signals with a relatively longer rise time. Other failure modes such as matrix failure and debonding produced lower amplitude signals with shorter rise times. We also noted that few events were recorded during the initial period of tensile loading (the elastic phase). The activity then increased significantly after maximum load was reached. The location information provided by the acoustic emission system was consistent with the final site of rupture. We have shown that AE can be used to characterise ligament damage, with fibre pull-outs and fibre fracture producing the highest signal amplitudes. PMID- 11018460 TI - Classification of waist-acceleration signals in a continuous walking record. AB - We attempted to distinguish walking on level ground from walking on a stairway using waist acceleration signals. A triaxial accelerometer was fixed to the subject's waist and the three acceleration signals were recorded by a portable data logger at a sampling rate of 256 Hz. Twenty healthy male subjects were asked to walk through a corridor and up and down a stairway as a single sequence, without any instruction. The data were analyzed using discrete wavelet transform. Walking patterns were classified in two stages. In the first stage, the times when the walking pattern changed were detected using the low-frequency component of the anteroposterior acceleration (LF(A)) and of the vertical acceleration (LF(V)). In the second stage, the three types of walking patterns were classified by comparing powers of wavelet coefficients in the vertical direction (P(WCV)) and in the anteroposterior direction (RP(WCA)). Changes in walking patterns could be detected by using either LF(A) or LF(V). Walking down stairs could be distinguished from the other types of walking as it gave the largest value in P(WCV), and walking up stairs could be discriminated from level walking using RP(WCA). Level and stairway walking could be classified from continuous records of waist acceleration. PMID- 11018461 TI - A new method for qualitative and quantitative evaluation of tooth displacement under the application of orthodontic forces using magnetic sensors. AB - The purpose of this study was to develop a new method for the quantitative and qualitative measurement of tooth displacement under load and to clarify the relationship between the force system acting on a tooth and a resultant movement. A combination of eight magnetic sensors and a magnet was employed to measure three-dimensional displacement. Two sets of sensors and a magnet were placed in front and at the back of a front tooth subjected to orthodontic forces. Magnets and sensors were fixed to the subject tooth and posterior teeth, respectively. Sensors detected displacements of two magnets extended from the subject tooth separately in real time. The tooth trajectories projected on the sagittal plane were studied. From displacements of two magnets, angle of tooth rotation, the position of center of rotation and the amount and direction of displacement of an arbitrary point on the tooth could be calculated with high accuracy. Also, the location of center of resistance could be determined. It was found that the center of rotation is moving substantially even with a small difference in the level of force application. Furthermore, a small amount of displacement of the center of resistance was observed with varying the force magnitude. PMID- 11018462 TI - Speeding-up the cure of one-part silicone rubber, when encapsulating neurological prostheses: the permeable mould. AB - The Note describes a procedure for encapsulating the implantable microelectronics in an air-curing silicone rubber, by which the shape of the casting is fully defined, and cure is achieved within an acceptable time. PMID- 11018463 TI - Temperature distribution for combined laser hyperthermia-photodynamic therapy in the esophagus. AB - In recent years photodynamic laser therapy (PDT) has been tested in animal and clinical studies for treatment of esophageal cancer. In several animal experiments a synergistic effect was found by simultaneously applying PDT and hyperthermia (HT). In this paper an optical fibre system is described which can be used in the esophagus for combined PDT with a 1 W dye laser and HT with a 15 W Nd:YAG laser. A phantom was built simulating the geometry of the esophagus using cow muscle. The spatial temperature field during HT was measured versus irradiation time. The results were compared with calculations using a coupled Monte Carlo laser transport/finite difference heat transport model using the LATIS computer program. Measurements and calculations yield a realistic description of the temperature distribution during HT under various experimental conditions. The LATIS program allows the prediction of the effects of blood perfusion for in vivo situations. The results show that perfusion has considerable influence on the temperature field, reducing the effective depth in tissue for HT. PMID- 11018464 TI - Chlorophyllin as an effective antioxidant against membrane damage in vitro and ex vivo. AB - Chlorophyllin (CHL), the sodium-copper salt and the water-soluble analogue of the ubiquitous green pigment chlorophyll, has been attributed to have several beneficial properties. Its antioxidant ability, however, has not been examined in detail. Using rat liver mitochondria as model system and various sources for the generation of reactive oxygen species (ROS) we have examined the membrane protective properties of CHL both under in vitro and ex vivo conditions. Oxidative damage to proteins was assessed as inactivation of the enzymes, cytochrome c oxidase and succinic dehydrogenase besides formation of protein carbonyls. Damage to membrane lipids was measured by formation of lipid hydroperoxides and thiobarbituric acid reactive substances. The effect of this compound on the antioxidant defense system was studied by estimating the level of glutathione and superoxide dismutase. ROS were generated by gamma-radiation, photosensitization, ascorbate-Fe(2+), NADPH-ADP-Fe(3+) and the peroxyl radical generating agent, azobis-amidopropane hydrochloride. Our results show that CHL is highly effective in protecting mitochondria, even at a low concentration of 10 microM. The antioxidant ability, at equimolar concentration, was more than that observed with ascorbic acid, glutathione, mannitol and tert-butanol. When CHL was fed to mice at a dose of 1% in drinking water, there was a significant reduction in the potential for oxidative damage in cell suspensions from liver, brain and testis. To examine the possible mechanisms responsible for the observed antioxidant ability we have studied the reaction of CHL with the potent ROS in the form of hydroxyl radical and singlet oxygen. The compound shows a fairly high rate constant with singlet oxygen, in the order of 1.3x10(8) M(-1) s(-1). In conclusion, our studies showed that CHL is a highly effective antioxidant, capable of protecting mitochondria against oxidative damage induced by various ROS. PMID- 11018465 TI - Human sphingosine-1-phosphate lyase: cDNA cloning, functional expression studies and mapping to chromosome 10q22(1). AB - Sphingosine-1-phosphate lyase catalyzes the last step in sphingolipid breakdown, the cleavage of phosphorylated sphingoid bases such as sphingenine-1-phosphate. The latter lipid is not only a catabolite, but can influence as an inter- and/or intracellular second messenger many cellular processes. To allow for the diagnosis of human disorders that might be linked to a deficient lyase, the human sphingosine-1-phosphate lyase cDNA was cloned. The obtained cDNA encoded a protein of 568 amino acids with a calculated molecular mass of 63492 Da. Hydropathy plots revealed the presence of one membrane span near the amino terminal which is however not required for enzyme activity since recombinant poly His-tagged lyase, lacking this membrane span, was functionally active. Site directed mutagenesis disclosed the importance of the cysteine residues 218 and 317 for the cleavage reaction. Northern analysis showed the presence of rare large-sized mRNAs of 6.7, 5.8 and 4 kb and the highest expression in liver. By fluorescent in situ hybridization, the gene was mapped to chromosome 10q22. PMID- 11018466 TI - Docosahexaenoic acid-deficient phosphatidyl serine and high alpha-tocopherol in a fetal mouse brain over-expressing Cu/Zn-superoxide dismutase. AB - The over-expressed Cu/Zn-superoxide dismutase (Cu/Zn-SOD) gene has been found in some circumstances phenotypically deleterious and associated with oxidative injury-mediated aberrations while in other studies it was considered neuroprotective. In this work we examine a number of biochemical markers in fetal and adult brain from transgenic (tg) mice expressing the human Cu/Zn-SOD gene, which may determine this dual characteristic. These markers include the polyunsaturated fatty acid (PUFA) profile in discrete phospholipid species, the alpha-tocopherol levels, a marker for lipid anti-oxidant status, and thiobarbituric acid reactive substance (TBARS), a marker for the tissue oxidative status. The PUFA profile in choline- and ethanolamine-phosphoglycerides was similar in tg and nontransgenic (ntg) animals of either fetal or adult brain. Serine-phosphoglycerides, however, showed a marked decrease from 20. 07+/-0.53 to 14.92+/-0.87 wt% and 14.52+/-1.15 wt% in docosahexaenoic acid (DHA; 22:6 n3), in the tg 51 and tg 69 fetal brains, respectively, but not in the comparable adult tissues. The alpha-tocopherol levels were significantly higher in the fetal compared to the adult brain. There were no differences in the anti-oxidant levels between the ntg and tg fetal brains, but there were differences in the adult animals; the tg mice were higher by at least two-fold than the control animals. The basal TBARS in the tg 51 fetal brain was 35% lower than that of ntg mouse and in the presence of Fe(2+), brain slices from the former released less TBARS (57% reduction) into the medium than the latter. These results suggest that higher dosages of Cu/Zn-SOD gene are compatible with increased alpha-tocopherol levels, reduced basal TBARS levels and a DHA deficiency in the fetal, but not the adult, tg brain. PMID- 11018467 TI - Accumulation of cholestatic lipoproteins in ANIT-treated human apolipoprotein A-I transgenic rats is diminished through dose-dependent apolipoprotein A-I activation of LCAT. AB - Administration of alpha-naphthylisothiocyanate (ANIT) to rats induces changes to plasma lipids consistent with cholestasis. We have previously shown (J. Lipid Res. 37 (1996) 1086) that animals treated with ANIT accumulate large amounts of free cholesterol (FC) and phospholipid (PL)-rich cholestatic lipoproteins in the LDL density range by 48 h. This lipid was cleared by 120 h through apparent movement into HDL with concomitant cholesteryl ester (CE) production. It was hypothesised that the clearance was mediated through the movement of the PL and FC into apolipoprotein A-I (apo A-I) containing lipoproteins followed by LCAT esterification to form CE. To test this hypothesis, rats overexpressing various amounts of human apo A-I (TgR[HuAI] rats) were treated with ANIT (100 mg/kg) and the effect of plasma apo A-I concentration on plasma lipids and lipoprotein distribution was examined. In untreated TgR[HuAI] rats, human apo A-I levels were strongly correlated to plasma PL (r(2)=0. 94), FC (r(2)=0.93) and CE (r(2)=0.90), whereas in ANIT-treated TgR[HuAI] rats, human apo A-I levels were most strongly correlated to CE levels (r(2)=0.80) and an increased CE/FC ratio (r(2)=0.62) and the movement of cholestatic lipid in the LDL to HDL. Since LCAT activity was not affected by ANIT treatment, these results demonstrate that the ability of LCAT to esterify the plasma FC present in cholestatic liver disease is limited by in vivo apo A-I activation of the cholestatic lipid and not by the catalytic capacity of LCAT. PMID- 11018468 TI - Difference in substrate specificity between human and mouse lysosomal acid lipase: low affinity for cholesteryl ester in mouse lysosomal acid lipase. AB - Lysosomal acid lipase (LAL) is essential for the intracellular degradation of cholesteryl esters (CE) and triacylglycerols (TG) that are delivered to lysosomes by low density lipoprotein (LDL) receptor mediated endocytosis. We have analysed the difference in the catalytic properties and substrate specificity of human and mouse LALs. LAL activities were measured in human and mouse fibroblasts and in HeLa cells transiently expressing wild-type or site-directed mutant LALs of the two species using the T7 vaccinia system. Cholesteryl esterase and triacylglycerol lipase activities were determined in cellular homogenates with a phospholipid/detergent vesicle assay, an assay frequently used to diagnose human LAL deficiency syndromes, and with LDL particles, a more physiological substrate. Characterisation of human and mouse LAL using these two assays demonstrated marked differences in their TG and CE hydrolysing activities. Compared to human LAL mouse LAL showed a much lower cholesteryl esterase activity in both assays used. The difference was more pronounced in the vesicle assay. The lower cholesteryl esterase activity of mouse LAL did not affect the LDL-CE degradation in intact fibroblasts. The analysis of site-directed mutants suggests a role of the non-conserved cysteine residue at position 240 in cholesteryl esterase activity in human LAL. Our results show a significant difference between human and mouse LAL in their specificity toward cholesteryl esters. The low cholesteryl esterase activity does not result in reduced LDL-cholesterol ester degradation in mouse fibroblasts in situ. In addition, this work emphasises the importance of the physical state of substrates in studies of the specificity and properties of lipolytic enzymes. PMID- 11018469 TI - Expression of a peptide binding to receptor for activated C-kinase (RACK1) inhibits phorbol myristoyl acetate-stimulated phospholipase D activity in C3H/10T1/2 cells: dissociation of phospholipase D-mediated phosphatidylcholine breakdown from its synthesis. AB - The C3H/10T1/2 Cl8 HAbetaC2-1 cells used in this study express a peptide with a sequence shown to bind receptor for activated C-kinase (RACK1) and inhibit cPKC mediated cell functions. Phorbol myristoyl acetate (PMA) strongly stimulated phosphatidylcholine (PtdCho)-specific phospholipase D (PLD) activity in the C3H/10T1/2 Cl8 parental cell line, but not in Cl8 HAbetaC2-1 cells, indicating that full PLD activity in PMA-treated Cl8 cells is dependent on a functional interaction of alpha/betaPKC with RACK1. In contrast, the PMA-stimulated uptake of choline and its subsequent incorporation into PtdCho, were not inhibited in Cl8 HAbetaC2-1 cells as compared to Cl8 cells, indicating a RACK1-independent but PKC-mediated process. Increased incorporation of labelled choline into PtdCho upon PMA treatment was not associated with changes of either CDP-choline: 1,2 diacylglycerol cholinephosphotransferase activity or the CTP:phosphocholine cytidylyltransferase distribution between cytosol and membrane fractions in Cl8 and Cl8 HAbetaC2-1 cells. The major effect of PMA on the PtdCho synthesis in C3H/10T1/2 fibroblasts was to increase the cellular uptake of choline. As a supporting experiment, we inhibited PMA-stimulated PtdH formation by PLD, and also putatively PtdH-derived DAG, in Cl8 cells with 1-butanol. Butanol did not influence the incorporation of [(14)C]choline into PtdCho. The present study shows: (1) PMA-stimulated PLD activity is dependent on a functional interaction between alpha/betaPKC and RACK1 in C3H/10T1/2 Cl8 fibroblasts; and (2) inhibition of PLD activity and PtdH formation did not reduce the cellular uptake and incorporation of labelled choline into PtdCho, indicating that these processes are not directly regulated by PtdCho-PLD activity in PMA-treated C3H/10T1/2 Cl8 fibroblasts. PMID- 11018470 TI - Activation of phospholipase D by PKC and GTPgammaS in human neuroblastoma cells overexpressing MARCKS. AB - Regulation of phospholipase D (PLD) activity participating in signal transduction involves complex interactions with small G-proteins (ARF, Rho) and protein kinase C isoforms (PKCalpha). In SK-N-MC human neuroblastoma cells, phorbol ester (TPA) activation of PLD was enhanced by overexpressing myristoylated alanine-rich C kinase substrate (MARCKS). To study MARCKS interactions with PLD, we investigated PLD isoform expression and activation by TPA and GTPgammaS in intact and digitonin-permeabilized clones transfected with MARCKS (M22). PLD2 was in both cytosol and membrane fractions while PLD1 was primarily membrane-associated in both vector control and M22 cells; location or quantities were unaltered by TPA treatment. TPA-stimulated PLD activity was higher in both intact and digitonin permeabilized M22 cells than in vector controls. In contrast, GTPgammaS stimulated PLD activity was independent of MARCKS expression but was additive with MARCKS-PKC-dependent activation in permeabilized cells. Combinations of PKC inhibition and down-regulation in intact and permeabilized (with GTPgammaS present) cells indicated that a PKC-mediated phosphorylation event was necessary in intact cells without access to GTPgammaS, stimulation of PLD mediated by GTPgammaS was independent of PKC, and PLD activation by PKC in permeabilized cells was kinase-independent. Western blot analysis showed that MARCKS, PKCalpha, PLD1 and PLD2 were present in a detergent-insoluble fraction (DIF); GTPgammaS increased recovery of PLD2 in DIF. Disruption of cholesterol-rich DIFs with digitonin, cyclodextrin or filipin potentiated activation of PLD by TPA. Our studies suggest that activation of PLD by PKC requires MARCKS and can involve both phosphorylation-independent and -dependent processes. As PLD activation by GTPgammaS is PKC-MARCKS-independent, MARCKS may provide a fine tuning component in conjunction with G-protein-mediated mechanisms for regulation of PLD. PMID- 11018471 TI - Augmented resistance to oxidative stress in fatty rat livers induced by a short term sucrose-rich diet. AB - Hepatic steatosis and the accompanying oxidative stress have been associated with a variety of liver diseases. It is not known if fat accumulation per se plays a direct role in the oxidative stress of the organ. This study tested if steatosis induced by a short-term carbohydrate-rich diet results in an increased hepatic sensitivity to oxidative stress. Antioxidant status was determined in a liver perfusion system and in isolated parenchymal, endothelial and Kupffer cells from rats kept on sucrose-rich diet or on regular diet for 48 h. t-Butyl hydroperoxide addition (2 mM) to the perfusion fluid resulted in a release of alanine aminotransferase (ALT) in livers from controls, whereas no ALT release was observed in fatty livers. After t-butyl hydroperoxide addition, oxidized glutathione release was 40% less in fatty than in control livers, whereas reduced glutathione (GSH) release was not different. Sinusoidal oxidant stress was mimicked by the addition of lipopolysaccharide (LPS) from Escherichia coli (10 microg/ml) followed by the addition of opsonized zymosan (8 mg/ml) to the perfusion medium. LPS plus zymosan treatments resulted in the release of ALT in control but not in fatty livers. At the end of perfusion, liver glutathione content was 3-fold elevated, and the tissue content of lipid peroxidation products was approx. 40% less in fatty livers compared to controls. GSH content was doubled and glucose-6-phosphate dehydrogenase (G6PD) expression was elevated by 3- and 10-fold in sinusoidal endothelial and parenchymal cells form fatty livers compared to cells from control animals. Following H(2)O(2) administration in vitro (0.2-1 mM), GSH remained elevated in endothelial and parenchymal cells from fatty livers compared to cells from controls. In contrast, G6PD activity and GSH content were similar in Kupffer cells isolated from fatty or control livers. The study shows that hepatic fat accumulation caused by a short-term sucrose diet is not accompanied by elevated hepatic lipid peroxidation, and an elevated hepatic antioxidant activity can be manifested in the presence of prominent steatosis. The diet-induced increase in G6PD expression and, thus, the efficient maintenance of reduced glutathione in endothelial and parenchymal cells are a supportive mechanism in the observed hepatic resistance against intracellular or sinusoidal oxidative stress. PMID- 11018472 TI - Phosphatidylcholine-specific phospholipase C inhibitor, tricyclodecan-9-yl xanthogenate (D609), increases phospholipase D-mediated phosphatidylcholine hydrolysis in UMR-106 osteoblastic osteosarcoma cells. AB - Our previous studies have shown that parathyroid hormone (PTH) stimulates phosphatidylcholine (PC) hydrolysis by phospholipase D (PLD) and transphosphatidylation in UMR-106 osteoblastic cells. To determine whether phospholipase C (PLC) is also involved in the PTH-mediated PC hydrolysis, we used the inhibitor, tricyclodecan-9-yl xanthogenate (D609), a putatively selective antagonist of this pathway. Consistent with this proposed mechanism, D609 decreased (3)H-phosphocholine in extracts from UMR-106 cells prelabeled with (3)H choline. Unexpectedly, D609 enhanced PC hydrolysis and transphosphatidylation, suggesting that either there was a compensatory increase in PLD activity when PLC was inhibited, or that D609 directly increased PLD activity. The D609-stimulated increase in PC hydrolysis was rapid, being seen as early as 2 min. The effect of D609 was temperature-sensitive, consistent with an enzymatic mechanism. The D609 stimulated increase in PC hydrolysis was PKC-independent, based upon the lack of effect of down-regulation of PKC by phorbol 12,13-dibutyrate on the response. The studies reveal a novel action of this inhibitor on signaling in osteoblastic cells which might influence downstream responses. PMID- 11018473 TI - Phosphatidylinositol-specific phospholipase C-gamma1 undergoes pH-induced activation and conformational change. AB - Phospholipase C-gamma1 displayed sigmoidal kinetics with a S(0.5) value of 0.17 mole fraction PIP(2) when assayed at pH 6.8 using detergent:lipid mixed micelles. The pH optimum for hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase C-gamma1 was dependent on the mole fraction of substrate in the micelle. The pH optimum was 5.5 when the enzyme was assayed below the S(0.5). The pH optima shifted to a pH range of 6.0-6.3 when the enzyme was assayed above the S(0.5). The kinetic parameters for phospholipase C-gamma1 assayed at various pH values from pH 7.0 to 5.0 yielded similar n values (n=4), but the constant, K', decreased from 1x10(-2) (mole fraction)(2) at pH 7.0 to 1x10(-5) (mole fraction)(2) at pH 5.0. Maximum enzyme specificity occurred at pH values below pH 6.0 as determined by the plot of logk(cat)/S(0.5) versus pH. Intrinsic fluorescence spectroscopy revealed that at a pH value above 7.0 or below 6.3, tryptophan quenching occurred. Fluorescence quenching experiments performed with acrylamide determined phospholipase C-gamma1 incubated at pH 5.0 had a larger collisional quenching constant than enzyme incubated at pH 7.0. Lowering the pH to 5.0 apparently resulted in interior tryptophans becoming more solvent accessible. These data suggest that pH may activate phospholipase C-gamma1 by disrupting ionizable groups leading to a conformational change. PMID- 11018474 TI - Characterization of 4-hydroxy-2-nonenal metabolism in stellate cell lines derived from normal and cirrhotic rat liver. AB - During oxidative stress, reactive aldehydes, including trans-4-hydroxy-2-nonenal (4-HNE), are generated by peroxidation of membrane lipids and purportedly stimulate hepatic stellate cells to produce excessive extracellular matrix, including type I collagen. An important question concerning the ability of 4-HNE to modulate collagen production by stellate cells is the potential of these specialized cells to detoxify 4-HNE. The objective of the present study was to characterize the ability of stellate cell lines, derived from normal (NFSC) and cirrhotic (CFSC) rat livers, to metabolize 4-HNE by oxidative, reductive and conjugative pathways. These two stellate cell lines were noted to have differing susceptibilities to the cytotoxic effect of 4-HNE. Treatment of both stellate cell lines with a range of 4-HNE doses demonstrated that the concentration which was cytotoxic to 50% of CFSC (TD(50)) was 25% greater than that for NFSC (967.57+/-9.26 nmol/10(6) cells vs. 769.90+/-5.32 nmol/10(6) cells respectively). The capacity of these cell lines to metabolizes 4-HNE was determined by incubating them in suspension with 50 microM 4-HNE (10 nmol/10(6) cell); 4-HNE elimination and metabolite formation were quantified over a 20 min time course. Both stellate cell lines rapidly metabolized 4-HNE, with the CFSC line eliminating 4-HNE at a rate that was approx. 2-fold greater than the NFSC line. The rate of 4-HNE metabolism attributable to glutathione S-transferase (GST) was similar in both cell lines, though differential cell specific expressions of GST isoforms GSTP1-1 and GSTA4-4 were observed. The greater rate of 4-HNE elimination by CFSC was attributable to its aldehyde dehydrogenase (ALDH) activity which accounted for approx. 50% of 4-HNE metabolism in CFSC but was insignificant in NFSC. Neither cell line had detectable alcohol dehydrogenase activity or protein levels. Measurement of cellular GSH concentrations revealed that NFSC contain approx. 2-fold greater concentrations of GSH when compared to CFSC and that following 4-HNE treatment, GSH levels were rapidly depleted from both cell lines. Concomitant with 4-HNE mediated GSH depletion, a corresponding increase in the 4 HNE-glutathione adduct formation was observed with the NFSC line forming greater amounts of the glutathione adduct than did the CFSC line. Taken together, these data demonstrate that both stellate cell lines have the capacity to metabolize 4 HNE but that CFSC have a greater rate of metabolism which is attributable to their greater ALDH activity, suggesting that the stellate cells isolated from cirrhotic liver may be differentially responsive to the biologic effects of 4 HNE. PMID- 11018475 TI - Lysosomal accumulation of oxidized phosphatidylcholine-apolipoprotein B complex in macrophages: intracellular fate of oxidized low density lipoprotein. AB - Oxidized phosphatidylcholine (OxPC) formed in oxidized low density lipoprotein (OxLDL) is thought to be involved in the development of atherosclerosis. OxPC has been found in foam cells in atherosclerotic lesions and suggested to be the epitope for OxLDL recognition by macrophages. OxPC is present as a complex with apolipoprotein B (apoB) in OxLDL, since some OxPC can bind with proteins. In the current study, the intracellular fate of OxPC-apoB complexes after internalization of OxLDL by macrophages was investigated. Murine macrophage cell line J774.1 was incubated with either OxLDL or acetylated LDL for 24 h, then the cells were further incubated for up to 24 h in new medium without lipoprotein. Modified apoB in the cells was quantitated by sandwich ELISA using monoclonal antibodies against OxPC and apoB. Intracellular OxLDL decreased rapidly for the first 4 h to approx. 20% of that before medium change, with the apparent metabolism of OxPC-apoB complex ceasing. OxPC-apoB complexes that remained in the cells after 24 h chasing increased as the period of OxLDL loading in macrophages prolongs. Acetylated LDL in the cells decreased quickly and disappeared after 4 h of chasing. Subcellular fractionation using sucrose density gradient ultracentrifugation of macrophages, which had already accumulated OxPC-apoB complexes by 24 h of incubation with OxLDL and further 24 h chasing, showed that the complex was co-localized with endosomal and lysosomal markers. Immunohistochemical double staining studies demonstrated that OxPC and apoB co localize in foam cells in early atherosclerotic lesions obtained from human coronary artery. These results suggest that OxPC-apoB complexes originating from OxLDL accumulate in foam cells in human atherosclerotic lesions as well as in macrophages in vitro. PMID- 11018476 TI - The C2 domain of protein kinase calpha is directly involved in the diacylglycerol dependent binding of the C1 domain to the membrane. AB - Protein kinase Calpha (PKCalpha), which is known to be critical for the control of many cellular processes, was submitted to site-directed mutagenesis in order to test the functionality of several amino acidic residues. Thus, D187, D246 and D248, all of which are located at the Ca(2+) binding site of the C2 domain, were substituted by N. Subcellular fractionation experiments demonstrated that these mutations are important for both Ca(2+)-dependent and diacylglycerol-dependent membrane binding. The mutants are not able to phosphorylate typical PKC substrates, such as histone and myelin basic protein. Furthermore, using increasing concentrations of dioleylglycerol, one of the mutants (D246/248N) was able to recover total activity although the amounts of dioleylglycerol it required were larger than those required by wild type protein. On the other hand, the other mutants (D187N and D187/246/248) only recovered 50% of their activity. These data suggest that there is a relationship between the C1 domain, where dioleylglycerol binds, and the C2 domain, and that this relationship is very important for enzyme activation. These findings led us to propose a mechanism for PKCalpha activation, where C1 and C2 domains cannot be considered independent membrane binding modules. PMID- 11018477 TI - Increased intestinal phospholipase A(2) activity catalyzed by phospholipase B/lipase in WBN/Kob rats with pancreatic insufficiency. AB - Male WBN/Kob rats derived from the Wistar strain spontaneously develop chronic pancreatitis as late as 3 months old. To assess the degree of disease severity, we compared the lipolytic enzyme levels in pancreas of 2-, 4-, and 6-month-old WBN/Kob rats fed isocaloric no fat (NF) and high fat (HF, 57% of total calories) diets and its pathology. Diet treatment did not significantly affect lipase and group Ib phospholipase A(2) (PLA(2)) levels in the pancreas at all ages. Development of chronic pancreatitis at the age of 4 and 6 months was consistent with the tendency of decreasing group Ib PLA(2) specific content determined by enzyme immunoassay and lipase activity, and the decreased number of group Ib PLA(2)-positive acinar cells. Pancreatic lipase and group Ib PLA(2) levels of 4 month-old WBN/Kob rats were significantly lower than those of control Wistar rats at age 4 months irrespective of diet. This allowed us to adopt 4-month-old WBN/Kob rats as a model of pancreatic insufficiency, which could be a useful tool to examine the role of gastrointestinal enzymes in lipid digestion. Ca(2+) independent PLA(2) activity of brush border membrane-associated phospholipase B/lipase (PLB/LIP) in ileal mucosa increased significantly in 4-month-old WBN/Kob rats while its content and transcript levels remained constant, suggesting its activation at the enzyme level. In WBN/Kob rats fed the HF diet at age 4 months, PLA(2) activity catalyzed by PLB/LIP in the proximal ileal mucosa was four times the total PLA(2) activity in the intestinal lumen. These results indicate that PLB/LIP compensates for the depletion of pancreatic lipolytic enzymes in WBN/Kob rats with pancreas insufficiency. PMID- 11018478 TI - Shedding of sulfated lipids into gastric fluid and inhibition of pancreatic DNase I by cholesterol sulfate in concert with bile acids. AB - Cholesterol sulfate (CS) and sulfatides in the epithelium of the digestive tract were found in the 1000xg supernatants of digestive fluid, particularly in gastric juices containing the duodenal contents and bile acids, there being 14-131 microg of CS and 3-54 microg of sulfatides per mg of protein in the fluid, respectively. CS and sulfatides dissolved in detergents including bile acids inactivated pancreatic trypsin to the same level as by DMSO-solubilized sulfated lipids at 37 degrees C. Similarly, pancreatic DNase I was inhibited by CS solubilized with DMSO or bile acids, but not by sulfatides or other membrane lipids at 37 degrees C. Both the sulfate group and the hydrophobic side chain of CS were indispensable structures for the inhibition of DNase I. Also, the optimum molar ratio of bile acids to CS was important for expression of the inhibitory activity of CS toward DNase I, it being 0.18 of the optimum ratio for sodium taurocholate, and the molar ratio of CS to DNase I for complete inhibition was 342:1. Thus, CS was shown to play a role as an epithelial inhibitor of DNase I in concert with bile acids. PMID- 11018479 TI - GDP-fucose: beta-galactoside alpha1,2-fucosyltransferase, MFUT-II, and not MFUT-I or -III, is induced in a restricted region of the digestive tract of germ-free mice by host-microbe interactions and cycloheximide. AB - A shift from sialylation to fucosylation of mucosal glycoconjugates occurred in the mammalian digestive tract in the weaning period, but mice under germ-free conditions were found to express both fucosyl GM1 (FGM1) and fucosyl asialo GM1 (FGA1) in the stomach, cecum and colon, but not in the small intestine. By host microbe interactions and administration of cycloheximide, FGA1 was quickly induced in the small intestine, but the concentrations of fucosylated glycolipids in the other regions were not altered significantly. Their expression coincided with the activity of GDP-fucose:GA1 alpha(1, 2)-fucosyltransferase (alpha1,2-FT), and we isolated a cDNA with an open reading frame encoding the murine alpha1,2-FT (MFUT-II) of 347 amino acids with a predicted molecular mass of 39.21 kDa. The intraperitoneal injection of cycloheximide induced the mRNA and activity of alpha1,2-FT (MFUT-II) in the small intestine of germ-free mice, whereas no change in the mRNA or activity was observed in the stomach, cecum and colon, indicating that expression of FGA1 in response to microbial colonization or cycloheximide is transcriptionally regulated in a restricted region of the murine digestive tract. At 24 h after the administration of cycloheximide, FGA1 was preferentially produced in the upper half of the duodenal microvilli. PMID- 11018480 TI - Lipid composition and dynamics of cell membranes of Bacillus stearothermophilus adapted to amiodarone. AB - Bacillus stearothermophilus, a useful model to evaluate membrane interactions of lipophilic drugs, adapts to the presence of amiodarone in the growth medium. Drug concentrations in the range of 1-2 microM depress growth and 3 microM completely suppresses growth. Adaptation to the presence of amiodarone is reflected in lipid composition changes either in the phospholipid classes or in the acyl chain moieties. Significant changes are observed at 2 microM and expressed by a decrease of phosphatidylethanolamine (relative decrease of 23.3%) and phosphatidylglycerol (17.9%) and by the increase of phosphoglycolipid (162%). The changes in phospholipid acyl chains are expressed by a decrease of straight-chain saturated fatty acids (relative decrease of 12.2%) and anteiso-acids (22%) with a parallel increase of the iso-acids (9.8%). Consequently, the ratio straight chain/branched iso-chain fatty acids decreases from 0. 38 (control cultures) to 0.30 (cultures adapted to 2 microM amiodarone). The physical consequences of the lipid composition changes induced by the drug were studied by fluorescence polarization of diphenylhexatriene and diphenylhexatriene-propionic acid, and by differential scanning calorimetry. The thermotropic profiles of polar lipid dispersions of amiodarone-adapted cells are more similar to control cultures (without amiodarone) than those resulting from a direct interaction of the drug with lipids, i.e., when amiodarone was added directly to liposome suspensions. It is suggested that lipid composition changes promoted by amiodarone occur as adaptations to drug tolerance, providing the membrane with physico-chemical properties compatible with membrane function, counteracting the effects of the drug. PMID- 11018481 TI - Preferential externalization of newly synthesized phosphatidylserine in apoptotic U937 cells is dependent on caspase-mediated pathways. AB - Externalization of phosphatidylserine (PtdSer) is a common feature of programmed cell death and plays an important role in the recognition and removal of apoptotic cells. In this study with U937 cells, PtdSer synthesis from [(3)H]serine was stimulated and newly synthesized PtdSer was transferred preferentially to cell-free medium vesicles (CFMV) from cells when apoptosis was induced with a topoisomerase I inhibitor, camptothecin (CAM). When CAM-induced apoptosis was blocked by a caspase inhibitor, z-VAD-fmk, stimulation of PtdSer synthesis and movement to CFMV were abolished. In contrast, changes in synthesis and transport of sphingomyelin (SM) or phosphatidylethanolamine (PtdEtn) were minor; total phosphatidylcholine (PtdCho) synthesis was below control levels. All phospholipids appeared in CFMV but PtdSer displayed a 6-fold increase relative to controls compared to 3-fold for SM, 2-fold for PtdCho and 1.8-fold for PtdEtn. Even greater effects on specificity of PtdSer synthesis, movement to CFMV and inhibition by z-VAD-fmk were observed in apoptotic cells induced by UV irradiation or tumor necrosis factor-alpha/cycloheximide treatment. Thus, PtdSer biosynthesis stimulated during apoptosis in U937 cells was specific for this phospholipid and was correlated with caspase-mediated exposure of PtdSer at the cell surface and preferential movement to vesicles during apoptosis. PMID- 11018482 TI - Induction of hepatic cytosolic fatty acid binding protein with clofibrate accelerates both membrane and cytoplasmic transport of palmitate. AB - The role of liver cytosolic fatty acid binding protein (L-FABP) in fatty acid transport and metabolism is unclear. Female liver contains substantially more L FABP than male liver. Female liver also has a different fatty acid transport phenotype, including more rapid uptake, efflux and cytoplasmic transport. However, it is not known if the greater levels of L-FABP are responsible for these differences. We therefore determined whether increasing L-FABP using clofibrate causes male liver to acquire a female transport phenotype. The multiple indicator dilution (MID) method was used to estimate the rate constants for influx, efflux and cytoplasmic diffusion of palmitate in isolated perfused rat livers. Clofibrate treatment increased cytosolic concentrations of L-FABP 4.2+/-0.8-fold, the rate of cytoplasmic diffusion of palmitate 4.3+/-1.7-fold, and the steady-state palmitate extraction 1.5+/-0.3-fold (mean+/-S.E.). Influx and efflux constants were both increased (by 44% and 79%, respectively) to levels typical of female livers. These data suggest that clofibrate-induced elevation of cytosolic L-FABP not only stimulates intracellular diffusion but also influx and efflux of fatty acids. Possible mechanisms include reducing fatty acid binding to cytoplasmic membranes, induction of membrane fatty acid carriers, and catalyzing fatty acid exchange between aqueous cytoplasm and the plasma membrane. PMID- 11018483 TI - Essential fatty acid deficiency affects the fatty acid composition of the rat small intestinal and colonic mucosa differently. AB - The intestinal mucosal fatty acid (FA) composition was investigated in Sprague Dawley rats after 7 and 23 weeks on an isocaloric diet with qualitatively different essential fatty acid (EFA) composition. For comparison, serum and red blood cell (RBC) membranes were investigated in parallel. The molar percentage of most FAs differed significantly between serum and RBC membranes both in controls and rats fed an EFA deficient (EFAD) diet. The influence of the EFA diet was similar on serum and RBC membrane phospholipids except for arachidonic acid (AA) which was more markedly decreased in serum than in RBC membranes. The FA composition was similar in ileal and colonic mucosa, markedly differing from the jejunal mucosa, in which the AA concentration was lower (13.0+/-0.8 versus 16.8+/ 0.5 and 15. 7+/-2.8 mol%) and the linoleic acid (LA) concentration higher (34. 0+/-1.6 versus 17.8+/-1.3 and 15.5+/-2.8 mol%, respectively). The EFAD diet induced a more than five-fold decrease in the jejunal and ileal concentration of LA from 33.9+/-1.6 to 6.0+/-1.5 mol% and 17. 8+/-1.3 to 2.1+/-0.7 mol%, respectively. AA decreased more in the ileal and colonic mucosa than in the jejunum. The changes in the FA composition of the intestinal compartments after EFAD diet were different from that in serum and RBC membranes, and did not further change after 23 weeks compared to 7 weeks after introduction of the diet. The study shows that dietary influences are tissue specific and serum or RBC membranes do not mirror local changes in any of the different intestinal segments. PMID- 11018484 TI - Relationships between attention effects and intensity effects on the cognitive N140 and P300 components of somatosensory ERPs. AB - OBJECTIVES: This study attempts to elucidate the relative contributions of exogenous and endogenous components to the N140 and P300 potentials elicited by somatosensory stimulation. METHODS: Somatosensory event-related potentials (ERPs) were evoked using an odd-ball paradigm with the frequent (80%) stimuli delivered to the left index finger and the infrequent (20%) stimuli delivered to the right index finger. Both types of stimuli had the same intensity within each experiment. The experiment was repeated using 6 different stimulus intensities ranging from the sensory threshold to 3 times the threshold. Each experiment was done under two conditions. In one, the subjects were asked to count and respond to the infrequent stimuli. In the other, the subjects were instructed to ignore the stimuli whether frequent or infrequent. In addition, the compound sensory potential of the right median nerve was separately recorded from electrodes at the wrist using the same range of stimulus intensities applied to the right index finger. RESULTS: Amplitudes of the N140 and P300 elicited by both attended and unattended infrequent stimuli increased in a parallel fashion as a function of stimulus intensity, so that the amplitude difference between attended and unattended responses was independent of the stimulus intensity. The amplitude of the compound sensory nerve potential at the wrist exhibited a similar slope to those of the N140 and P300. CONCLUSIONS: Thus, it is concluded that the scalp N140 and P300 consist of two components: an endogenous component, which is independent of the stimulus intensity, and an exogenous component, which increases as a function of stimulus intensity. The relative contribution of these components to the N140 and P300 amplitudes is also discussed. PMID- 11018485 TI - A new strategy involving multiple cognitive paradigms demonstrates that ERP components are determined by the superposition of oscillatory responses. AB - OBJECTIVES: The goal of the present paper was to study the contribution of the delta and theta responses to two components of the event-related potential (ERP) waveform, the N200 and P300, which were recorded from 3 topographical sites of the brain. METHODS: This contribution was studied using a set of systematically varying experimental paradigms. Such a strategy enabled the demonstration of the variations in the event-related potentials and the event-related oscillations as task conditions and respective cognitive operations systematically changed. The study employed easy oddball, hard oddball, mismatch negativity and single stimulus paradigms and it was conducted on 42 healthy adults (age range 19-30 years, 26 females, 16 males) from the university student population. Data were analyzed with electrophysiological (selective averaging, amplitude frequency characteristics, digital filtering) and statistical methods (analysis of variance, multivariate step-down regression). RESULTS: The data showed that the morphology of the ERP components for different experimental paradigms represented a specific pattern of superposition of the delta and theta oscillatory responses. CONCLUSIONS: The cognitive correlates of the oscillatory responses were discussed and the results were evaluated on the basis of the superposition principle and the theory of oscillatory neural assemblies. PMID- 11018486 TI - Associated deficits in mismatch negativity generation and tone matching in schizophrenia. AB - OBJECTIVE: Schizophrenia is associated with deficits in mismatch negativity (MMN) generation and in the ability to match two tones following brief delay. Both deficits reflect impaired early cortical processing of auditory information. However, the relationship between deficits in MMN generation and tone matching performance in schizophrenia has not been established. METHODS: MMN and tone matching performance was evaluated in 12 schizophrenic subjects and 12 similar aged controls. A pitch separation known to produce non-ceiling performance in patients (5% Deltaf) was used. Narrow band filtering of MMN data was used to enhance signal-to-noise ratio. RESULTS: Schizophrenic subjects showed impairments in both MMN generation and tone matching performance. The two deficits were significantly correlated across subjects. In addition, decreased MMN amplitude and poorer tone matching performance correlated with increased severity of negative symptoms. CONCLUSIONS: These findings support the concept that similar neurophysiological mechanisms underlie MMN and tone matching deficits in schizophrenia. Further, they indicate that increased sensitivity to environmental change may be related to social withdrawal and other negative symptoms in schizophrenia. PMID- 11018487 TI - Somatosensory cortex responses to median nerve stimulation: fMRI effects of current amplitude and selective attention. AB - OBJECTIVES: The aim of this study was to localize and to investigate response properties of the primary (SI) and the secondary (SII) somatosensory cortex upon median nerve electrical stimulation. METHODS: Functional magnetic resonance imaging (fMRI) was used to quantify brain activation under different paradigms using electrical median nerve stimulation in healthy right-handed volunteers. In total 11 subjects were studied using two different stimulus current values in the right hand: at motor threshold (I(max)) and at I(min) (1/2 I(max)). In 7 of these 11 subjects a parametric study was then conducted using 4 stimulus intensities (6/6, 5/6, 4/6 and 3/6 I(max)). Finally, in 10 subjects an attention paradigm in which they had to perform a counting task during stimulation with I(min) was done. RESULTS: SI activation increased with current amplitude. SI did not show significant activation during stimulation at I(min). SII activation did not depend on current amplitude. Also the posterior parietal cortex appeared to be activated at I(min). The I(min) response in SII significantly increased by selective attention compared to I(min) without attention. At I(max) significant SI activity was observed only in the contralateral hemisphere, the ipsilateral cerebellum, while other areas possibly showed bilateral activation. CONCLUSIONS: Distributed activation in the human somatosensory cortical system due to median nerve stimulation was observed using fMRI. SI, in contrast to SII, appears to be exclusively activated on the contralateral side of the stimulated hand at I(max), in agreement with the concept of SI's important role in processing of proprioceptive input. Only SII remains significantly activated in case of lower current values, which are likely to exclusively stimulate the sensible fibres mediating cutaneous receptor input. Selective attention only enhances SII activity, indicating a higher-order role for SII in the processing of somatosensory input. PMID- 11018488 TI - Removal of eye activity artifacts from visual event-related potentials in normal and clinical subjects. AB - OBJECTIVES: Electrical potentials produced by blinks and eye movements present serious problems for electroencephalographic (EEG) and event-related potential (ERP) data interpretation and analysis, particularly for analysis of data from some clinical populations. Often, all epochs contaminated by large eye artifacts are rejected as unusable, though this may prove unacceptable when blinks and eye movements occur frequently. METHODS: Frontal channels are often used as reference signals to regress out eye artifacts, but inevitably portions of relevant EEG signals also appearing in EOG channels are thereby eliminated or mixed into other scalp channels. A generally applicable adaptive method for removing artifacts from EEG records based on blind source separation by independent component analysis (ICA) (Neural Computation 7 (1995) 1129; Neural Computation 10(8) (1998) 2103; Neural Computation 11(2) (1999) 606) overcomes these limitations. RESULTS: Results on EEG data collected from 28 normal controls and 22 clinical subjects performing a visual selective attention task show that ICA can be used to effectively detect, separate and remove ocular artifacts from even strongly contaminated EEG recordings. The results compare favorably to those obtained using rejection or regression methods. CONCLUSIONS: The ICA method can preserve ERP contributions from all of the recorded trials and all the recorded data channels, even when none of the single trials are artifact-free. PMID- 11018489 TI - Perception of complex sounds: N1 latency codes pitch and topography codes spectra. AB - OBJECTIVES: This work aimed to find out whether the human cortical 'tonotopy' represents the true fundamental frequency (Fo) of complex sounds, or the center frequency CF at which harmonics peak in the audio spectrum. Indeed, complex periodic sounds (such as those of the human voice, musical instruments, etc.) comprise a 'fundamental component' (Fo) and its 'harmonics' (2Fo, 3Fo, ...nFo). These often peak around a certain frequency CF. As Fo and CF are confounded in pure (sinusoidal) tones, the question of whether Fo or CF is represented through tonotopy had been hitherto unresolved. METHODS: Whole-head recordings of brain electrical activity were obtained for 16 subjects submitted to an array of 9 different series of sounds (3 Fox3 CF). Electrophysiological data were analyzed separately for each sound and each subject with brain functional imaging and dipole reconstruction. RESULTS: Equivalent dipole sources of N1 components were, significantly for all subjects, more and more frontally oriented as CF increased, independently of Fo. CONCLUSIONS: Sounds are mapped in both the right and the left primary auditory cortices according to the spectral profiles of their harmonics (CF), rather than their fundamental frequencies (Fo). PMID- 11018490 TI - Excitability recovery curve of the sympathetic skin response in healthy volunteers and patients with palmar hyperhidrosis. AB - OBJECTIVES: Patients with primary palmar hyperhidrosis (PPH) might exhibit hyperexcitability of the reflex circuits involved in sweating. We hypothesized that this hyperexcitability could become evident in the study of the excitability recovery curve of the sympathetic sudomotor skin response (SSR). METHODS: In 10 patients with PPH and 10 healthy volunteers used as control subjects, we recorded the SSR in the palm of the right hand to pairs of median nerve electrical shocks separated by inter-stimuli intervals (ISIs) ranging from 0.5 to 3.5 s. The amplitude of the SSR generated by the second stimulus (SSR2) was expressed as a percentage of that generated by the first (SSR1), and compared between control subjects and patients for each ISI. RESULTS: None of the control subjects showed a recovery of the SSR for ISIs of 1.5 s or less. On the contrary, patients showed a statistically significant enhancement of the SSR excitability recovery curve, with onset of recovery at 1.5 s in 5 patients. Two patients showed a double peak response to single electrical stimulation and were not considered in the calculation of the SSR recovery curve. Mean excitability recovery percentages were larger in patients than in control subjects at ISIs of 2, 2.5 and 3 s. CONCLUSIONS: The enhancement of the SSR recovery curve in patients with PPH suggests hyperexcitability of the somatosympathetic polisynaptic pathway involved in sweating. This could partly underlie the pathophysiology of PPH. PMID- 11018491 TI - Oculomotor impairment after 1 night of total sleep deprivation: a dissociation between measures of speed and accuracy. AB - OBJECTIVES: The present study examined the effects of 40 h of sleep deprivation and of time-of-day on saccadic and smooth pursuit oculomotor performance. METHODS: Nine normal subjects slept for 3 consecutive nights in the laboratory (one adaptation, one baseline, one recovery). Baseline and recovery were separated by a period of 40 h of continuous wakefulness, during which subjects were tested every 2 h. Oculomotor performance assessed at the following hours: 10:00, 12:00, 14:00, 16:00, 18:00, 20:00, 22:00, of both the days preceding and following the sleep deprivation night, as well as at 24:00, 02:00, 04:00, 06:00 and 08:00 h during the deprivation period. RESULTS: Saccade latency increased and peak velocity decreased significantly during the post-deprivation day; saccadic accuracy was unaffected. As regards smooth pursuit performance, phase (a measure of accuracy) was not affected by sleep loss, while velocity gain significantly decreased during the day that followed the sleep deprivation night. Significant time-of-day effects on the considered oculomotor variables except saccadic accuracy were also found, indicating an overall performance impairment during the night. CONCLUSIONS: It is concluded that 40 h of sleep deprivation significantly impaired diurnal performance in pursuit and saccadic tasks. This performance worsening is limited to the measures of speed, while accuracy is not affected by sleep loss. A significant operational relevance of these results is suggested, since saccadic velocity has recently been found to be negatively correlated with simulator vehicle crash rates. PMID- 11018492 TI - A novel quantitative EEG injury measure of global cerebral ischemia. AB - OBJECTIVE: To develop a novel quantitative EEG (qEEG) based analysis method, cepstral distance (CD) and compare it to spectral distance (SD) in detecting EEG changes related to global ischemia in rats. METHODS: Adult Wistar rats were subjected to asphyxic-cardiac arrest for sham, 1, 3, 5 and 7 min (n=5 per group). The EEG signal was processed and fitted into an autoregressive (AR) model. A pre injury baseline EEG was compared to selected data segments during asphyxia and recovery. The dissimilarities in the EEG segments were measured using CD and SD. A segment measured was considered abnormal when it exceeded 30% of baseline and its duration was used as the index of injury. A comprehensive Neurodeficit Score (NDS) at 24 h was used to assess outcome and was correlated with CD and SD measures. RESULTS: A higher correlation was found with CD and asphyxia time (r=0.81, P<0.001) compared to SD and asphyxia time (r=0.69, P<0.001). Correlation with cardiac arrest time (MAP<10 mmHg) showed that CD was superior (r=0.71, P<0.001) to SD (r=0.52, P=0.002). CD obtained during global ischemia and 90 min into recovery correlated significantly with NDS at 24 h after injury (Spearman coefficient=-0.83, P<0.005), and was more robust than the traditional SD (Spearman coefficient=-0.63, P<0.005). CONCLUSION: The novel qEEG-based injury index from CD was superior to SD in quantifying early cerebral dysfunction after cardiac arrest and in providing neurological prognosis at 24 h after global ischemia in adult rats. Studying early qEEG changes after asphyxic-cardiac arrest may provide new insights into the injury and recovery process, and present opportunities for therapy. PMID- 11018493 TI - Spectral analysis of electroencephalogram during sleep-related apneas in pre-term and term born infants in the first weeks of life. AB - OBJECTIVES: A correlation between sudden infant death syndrome (SIDS) and apnea has been discussed in a variety of studies. We analyzed polysomnographic data in 51 infants and investigated correlations between the manifestation of apneas and changes in EEG pattern. METHODS: Measurements were performed by means of the commercial polysomnographic device 'ALICE III'. Spectral analysis of certain parts of the registered EEG was performed. RESULTS: During apnea a reduction of the EEG amplitude was seen. The extent of the decrease differed slightly between the frequency bands. CONCLUSIONS: According to the widespread agreement that a lower voltage of electrical brain activity is a fundamental sign of waking up, we hypothesize that the observed change might be an immature type of arousal reaction. PMID- 11018494 TI - Amplitude changes in the 8-12, 20-25, and 40 Hz oscillations in finger tremor. AB - OBJECTIVE: The study examined the amplitude and frequency modulation of the 8-12, 20-25, and 40 Hz frequencies of tremor to determine the degree to which increments of load affect the amplitude of these neural rhythms. METHODS: Finger acceleration from the middle phalange and electromyographic (EMG) activity of the extensor digitorum communis (EDC) muscle were recorded on 10 normal adult subjects. Two experiments are reported that manipulated loads ranging from 0 to 40 and 0 to 200 g that were attached to the distal portion of the outstretched middle phalange. RESULTS: There were 8-12, 20-25, and 40 Hz oscillations in the EMG recording but only the 8-12 and 20-25 Hz rhythms were present in the tremor and tremor-EMG coherence. Adding load to the finger reliably decreased the 20-25 Hz band of acceleration, reduced the relative power within the 20-25 Hz EMG band, increased the relative power of the 40 Hz band, but had no effect on the relative power within the 8-12 Hz EMG frequency band. The tremor-EMG coherence in the 8-12 and 40 Hz regions was independent of load, but was markedly reduced with load in the 20-25 Hz band. CONCLUSIONS: The 8-12, 20-25, and 40 Hz neural rhythms of physiological tremor have a stable frequency consistent with central oscillations. There is an increase in the relative power of the 40 Hz EMG band with force, but only the amplitude of the 20-25 Hz band is modulated by mechanical-reflex feedback. PMID- 11018495 TI - Interictal, unifocal spikes in refractory extratemporal epilepsy predict ictal origin and postsurgical outcome. AB - OBJECTIVES: To evaluate the significance of exclusively unifocal, unilateral, interictal epileptiform patterns on scalp electroencephalography (EEG) in surgical candidates with medically intractable extratemporal epilepsy. METHODS: We reviewed 126 patients with refractory extratemporal partial seizures who underwent epilepsy surgery at our center. All were followed for at least 2 years after resections. Surgery was based on ictal EEG recordings. We examined ictal onsets and surgical outcome in subjects whose preoperative, interictal scalp EEGs during long-term monitoring (LTM) demonstrated only unilateral, well-defined focal discharges, and outcome in patients whose interictal EEGs during LTM showed bilateral, non-localized, or multifocal epileptiform patterns. RESULTS: We found that 26 subjects exhibited only unilateral, unifocal, interictal epileptiform patterns. In all 26 cases (100%) clinical seizures arose from the regions expected by the interictal findings (P<0.0001, Sign test). At last follow-up 77% (20/26) of these patients were seizure-free, while 23% (6/26) had >75% reduction in seizures. This compares to the remaining patients, of whom 34% (34/100) were seizure-free, 41% (41/100) had >75% reduction in seizures, and 25% (25/100) had <75% reduction in seizures (P=0.0001, Fisher's Exact test). CONCLUSIONS: Strictly unifocal, interictal epileptiform patterns on scalp EEG, though seen in a minority of subjects, may be an important, independent factor in evaluating subjects with intractable extratemporal, localization-related epilepsy for surgical therapy. This finding is highly predictive of both ictal onsets and successful postsurgical outcome. PMID- 11018496 TI - Alteration of motor cortex excitability in response to intermittent photic stimulation. AB - OBJECTIVE: To measure the duration of cortical silent period (C-SP) in response to different frequencies of intermittent photic stimulation (IPS). METHODS: Ten normal volunteers without a history of seizures were studied. First, using a magnetic stimulator we determined the resting motor threshold (RMT). The C-SP produced by a single magnetic shock at 120% of RMT was measured in the maximally contracted abductor pollicis brevis. C-SP was then measured while subjects were exposed to IPS at 5, 10, 15, 20, 30, 50, and 60 Hz with closed eyes. RESULTS: The average duration of C-SP was 135+/-33. 2 ms with no photic stimulation. This did not change significantly when subjects were exposed to IPS at 5, 10, 15, 20, and 30 Hz. The duration of C-SP, however, decreased significantly at higher frequencies. At 50 Hz it was 116.2+/-43 ms (P=0.003) and at 60 Hz it was 112.7+/ 44.6 ms (P<0.001). CONCLUSION: Our findings suggest that motor cortex inhibition is altered in response to IPS at 50 and 60 Hz. PMID- 11018497 TI - Myoclonic seizures combined with partial seizures and probable pathophysiology of secondary bilateral synchrony. AB - OBJECTIVE: We report on a patient who showed an unusual transition from complex partial seizures to a combination of myoclonic seizures and partial seizures, and then to isolated myoclonic seizures. The pathophysiological mechanism of the myoclonic seizures in this uncommon condition was studied. METHODS: The interhemispheric small time differences were estimated in the ictal EEG discharges of myoclonic seizures for differentiation between primary and secondary bilateral synchrony. The estimation was performed by coherence and phase analysis based on the two-dimensional autoregressive model. RESULTS: The estimated interhemispheric time differences were at most 27.4 ms. The ictal activity of the myoclonic seizures was suggested to originate from a cortical focus in the right hemisphere, which was also the origin of the partial seizures. CONCLUSION: The patient was considered to have an unusual type of myoclonic seizures with a probable pathophysiological mechanism of secondary bilateral synchrony. PMID- 11018498 TI - Spatial pattern of cerebral glucose metabolism (PET) correlates with localization of intracerebral EEG-generators in Alzheimer's disease. AB - BACKGROUND: Since the measurement of human cerebral glucose metabolism (GluM) by positron emission tomography (PET) and that of human cerebral electrical activity by EEG reflect synaptic activity, both methods should be related in their cerebral spatial distribution. Healthy subjects do indeed demonstrate similar metabolic and neuroelectric spatial patterns. OBJECTIVE: The aim of the study was to show that this similarity of GluM and EEG spatial patterns holds true in a population with a high variability of glucose metabolism. METHODS: We investigated healthy control subjects and patients with varying degrees of cognitive dysfunction and varying GluM patterns by applying [18F]FDG PET and EEG. RESULTS: We demonstrated that the localization of intracerebral generators of EEG correlates with spatial indices of GluM. CONCLUSION: These results indicates that EEG provides similar spatial information about brain function as GluM-PET. Since EEG is a non-invasive technique, which is more widely available and can be repeated more often than PET, this may have important implications both for neuropsychiatric research and for clinical diagnosis. However, further studies are required to determine whether equivalent EEG dipole generators can yield a diagnostic specificity and sensitivity similar to that of GluM-PET. PMID- 11018499 TI - Determinants of physiologic tremor in a large normal population. AB - OBJECTIVES: It has been well established that peripheral mechanical resonant factors as well as central mechanisms may play a role in the generation of physiological tremor (PT). Furthermore it has been postulated that subject's attributes like age and sex might influence PT. The present study was designed to quantify these influences on PT in a large normal population. METHODS: Physiological hand and finger tremors were measured in a group of 117 normal subjects between 20 and 94 years of age using accelerometry and surface EMG recordings from the forearm flexor and extensor muscles. The hand tremor was measured in a postural position with and without weight, and the finger tremor was recorded with the arm outstretched, forearm supported and hand supported. Hand volume and grip force were measured in each subject. RESULTS: Hand tremor frequency (mean 7.7 Hz) was reduced significantly by added inertia (mean 5.2 Hz) and it was negatively correlated with hand volume while there was no correlation with grip force. Finger tremor showed, subject to the arm position, maximally 3 and at least two distinct frequency bands (1-4, 6-11 and 15-30 Hz) reflecting the resonance frequencies of the whole arm, the hand and the finger, respectively. A significant EMG peak was found in 50-80% of the recordings. This EMG synchronization gave rise to a corresponding accelerometer peak or a significant EMG-EMG coherence in about one-third of the population indicating a central component of PT because its frequency was unaffected by mechanical changes in the periphery. We did not find a significant influence of age on the tremor frequency, while the sex of the subjects slightly but significantly changed the frequency range of hand tremor. Multiple partial correlations revealed, however, that the only direct influence on hand tremor frequency is the hand volume indicating that the influence of sex on hand tremor frequency is an indirect effect produced by the significantly larger hands of male subjects. CONCLUSIONS: In conclusion, the main determinants of PT are the mechanical properties of the oscillating limb. Apart from the dominating peripheral resonance mechanism we found indications of an additional central component of PT in about one-third of the normal population. There was no age dependence of tremor frequency and it was shown that the influence of the subjects' sex on tremor frequency only represents an indirect mechanical effect. PMID- 11018500 TI - Vestibular rehabilitation outcomes: velocity trajectory analysis of repeated bench stepping. AB - OBJECTIVES: To quantify vestibular rehabilitation (VR) outcomes in patients with vestibulospinal reflex pathology (VSP) using a new technique, high curvature analysis (HCA), that measures space trajectory stability. METHODS: Twelve patients with VSP and 10 healthy controls performed a repeated stepping task in a motion analysis laboratory. Patients with VSP were tested before and after a 12 week VR program. Three dimensional whole body center of gravity (CG) was captured during repeated bench stepping trials (step up forward/step down backward), and the resulting horizontal plane CG velocity trajectories were analyzed using HCA. RESULTS: VSP patients were significantly less stable than healthy subjects prior to VR (P=0.022), but these differences disappeared following VR (P=0.148). The stability of the VSP patients during the stepping task at 3 months increased significantly from baseline values (P=0.013). CONCLUSIONS: Evaluating the efficacy of VR requires analysis techniques that are sensitive to changes in stability. We conclude that HCA can be used for assessing improvements in CG stability of VSP patients while performing a repeated bench stepping activity. PMID- 11018501 TI - Effects of somatosensory input on central fatigue: a pilot study. AB - OBJECTIVE: Depression of motor evoked potentials (MEPs) following transcranial magnetic stimulation (TMS) may be a sign of central motor fatigue. As a pilot study, we have examined whether post-exercise MEP depression can be compensated by application of sensory stimuli prior to TMS. METHODS: We studied 15 healthy volunteers (aged 21-28 years) who were required to perform an exercise protocol of ankle dorsiflexion until force fell below 66% of maximum force. MEPs were recorded from the right tibialis anterior muscle. Prior to TMS, electrical stimuli were applied to the ipsilateral sural nerve with an individual interstimulus interval between 50 and 80 ms. RESULTS: MEP areas decreased after exercise. When a sensory stimulus was administered MEPs did not change. CONCLUSION: We conclude that the effects of central fatigue may be influenced by application of sensory stimuli. PMID- 11018502 TI - High resolution spatiotemporal analysis of the contingent negative variation in simple or complex motor tasks and a non-motor task. AB - OBJECTIVES: Since the characteristics of the Bereitschaftspotential (BP) - voluntary movement paradigm of internally-driven movements - have been established recently by our group using high resolution DC-EEG techniques, it was of great interest to apply similar techniques to the other slow brain potential- contingent negative variation (CNV) of externally-cued movements--with the same motor tasks using the same subjects. METHODS: The CNV for simple bimanual sequential movements (task 1), complex bimanual sequential movements (task 2) and a non-motor condition (task 3) was recorded on the scalp using a 64 channel DC EEG in 16 healthy subjects, and the data were analyzed with high resolution spatiotemporal statistics and current source density (CSD). RESULTS: (1) The CNV was distributed over frontal, frontocentral, central and centroparietal regions; a negative potential was found at the frontal pole and a positive potential was found over occipital regions. (2) CNV amplitudes were higher for task 2 than for task 1, and there was no late CNV for task 3. (3) A high resolution spatiotemporal analysis revealed that during the early CNV component, statistical differences existed between the motor tasks (tasks 1 and 2) and the non-motor task (task 3), which occurred at frontocentral, central, centroparietal, parietal and parieto-occipital regions. During the late CNV component, additional significant differences were found not only between the motor tasks and the non motor task but also between motor task 1 and task 2 at frontocentral, central and centroparietal regions. (4) Comparison of the CNV between the frontomesial cortex (situated over the supplementary/cingulate areas, SCMA) and both lateral pre central areas (situated over the primary motor areas, MIs) showed that there was no statistically significant difference between the two cortical motor areas except for the early CNV. (5) Comparison of the CNV between the 3 tasks over the cortical motor areas showed that there were significant differences between the motor tasks and the non-motor task regarding the auditory evoked potential (AEP) and the early CNV component, and between all 3 tasks in the late CNV, the visual evoked potential (VEP(2)) and the N-P component. (6) The ranges and the densities of the CSD maps were larger and higher for complex than for simple tasks. The current sinks of the AEP and the early CNV were located at Fz, the late CNV at FCz and surrounding regions. As to be expected, current sources of the VEPs were located at the occipital lobes. The CNV was a current sink (negative) except for the VEP's main component which was a current source (positive). CONCLUSIONS: (1) The CNV topography over the scalp varied with the complexity of motor tasks and between motor and non-motor conditions. (2) The origin of the early CNV may rest in the frontal lobes, while the late CNV may stem from more extensive cortical areas including SCMA, MIs, etc. (3) The late CNV component is not identical with the BP. PMID- 11018503 TI - Motor recovery following stroke: a transcranial magnetic stimulation study. AB - OBJECTIVES: To verify the usefulness of early recording of motor evoked potentials (MEPs) in predicting motor outcome after stroke and to investigate the neural mechanisms underlying functional recovery following stroke. METHODS: We performed a comparative analysis of the behaviour of motor responses evoked by transcranial magnetic stimulation (TMS) of the ipsilateral and contralateral motor cortex in the affected and unaffected thenar muscles of 21 consecutive patients with acute stroke. RESULTS: According to the behaviour of MEPs in the affected muscles, patients could be divided into 3 groups: (a) 10 subjects with absent responses to TMS of both the damaged and undamaged hemisphere, whose motor recovery was poor and related to the size of MEPs on the normal side; (b) 5 subjects with larger MEPs upon TMS of the ipsilateral (undamaged) than of the contralateral (damaged) cortex, whose good recovery possibly resulted from the emergence of ipsilateral pathways; (c) 6 subjects with larger MEPs in the affected than in the unaffected muscles, whose good recovery was possibly subserved by alternative circuits taking over cortical deafferentation. CONCLUSIONS: Early MEP recording in acute stroke provides useful information on the clinical prognosis and the different mechanisms of motor recovery. PMID- 11018504 TI - Modification of the silent period by double transcranial magnetic stimulation. AB - OBJECTIVES: To study the time course of the changes of the inhibitory network of the human motor system, we investigated the silent period (SP) in 7 healthy subjects by double suprathreshold transcranial magnetic stimulation (TMS). METHODS: SPs and motor evoked potentials (MEPs) were recorded from the voluntarily activated right abductor digiti minimi muscle. Conditioning and test stimuli were delivered with equal intensity, which was set to yield a baseline SP duration of 130 ms by a single pulse, and with various interstimulus intervals (ISIs). In addition, a control experiment with adjustment of the intensity of single stimuli was performed. RESULTS: At ISIs of 20 and 30 ms the test pulse SP duration was prolonged, without increasing the MEP amplitude. The SP duration shortened at longer ISIs and showed a significant depression between ISIs of 60 110 ms. The shortened SP was accompanied by a diminished MEP. The control experiment revealed that the SPs evoked by the adjusted pulses were significantly shorter than the test pulse SPs. CONCLUSIONS: A conditioning stimulus can prolong and shorten the test pulse SP duration at different ISIs. The prolongation is probably cortically generated, whereas the shortening is likely to occur at a cortical and spinal level. PMID- 11018505 TI - Functional dissociation of lower and upper frequency mu rhythms in relation to voluntary limb movement. AB - OBJECTIVE: The goal of this study is to investigate the reactivity of central rhythms in the alpha band during self-paced voluntary finger and foot movement and to give an answer to the question, whether different types of mu rhythms exist. METHODS: The effect of self-paced, voluntary finger and foot movement was studied in a group of 12 right-handed healthy volunteers. The EEG was recorded from a grid of 34 electrodes placed over sensorimotor areas with inter-electrode distances of approximately 2.5 cm. The event-related desynchronization (ERD) was quantified in the 8-10 and 10-12 Hz bands. RESULTS: Both frequency components are blocked prior to and during movement and therefore, they have to be considered as mu rhythms. The lower frequency component results in a widespread movement-type non-specific ERD pattern, whereas the upper frequency component shows a more focused and movement-type specific pattern, clearly different with finger and foot movement. CONCLUSIONS: The distinct reactivity patterns provide evidence for the existence of two types of mu rhythms, a somatotopically non-specific lower frequency mu rhythm and a somatotopically specific mu rhythm characteristically found in the upper alpha frequency band. PMID- 11018506 TI - Autonomic nervous function assessment using thermal reactivity of microcirculation. AB - There are only a few reliable objective methods of diagnosing peripheral neuronal damage suitable for routine use; the most important is based on measurement of nerve conduction velocity, which only shows changes when severe disturbances are already present. However, it is precisely at this stage that the possibilities of therapy are no longer satisfactory. As small fibres are affected earlier in the course of most forms of PNP than the large ones, assessment of afferent as well as efferent C-fibre function gains importance in the management of this widespread disease. In assessment of autonomic dysfunction, variability of the heartbeat with deep breathing or the Valsalva manoeuvre is a good and generally accepted test, although not strongly associated with other PNP test abnormalities. However, axonal degeneration starts in the most distal parts of the axon due to impaired axonal transport. Therefore, the longest C-fibres, i.e. in the lower extremities, are affected first, and incipient changes are most prominent there. For this reason HLDF, a reflex response of the skin blood flow stimulated by heat, has advantages in assessment of early C-fibre dysfunction. Considering the fact that the afferent and efferent sympathetic C-fibres are involved in regulation of microcirculation, the skin blood flow regulation is investigated by means of laser Doppler flowmetry. The microcirculation is stimulated by heat and the reaction of microcirculation is assessed as a value for the function of afferent and efferent (sympathetic) C-fibres. The results of this method are in close correlation with electrophysiologic tests, which is not achieved with sudomotor function. PMID- 11018507 TI - The development of a software program for analyzing spontaneous and externally elicited skin conductance changes in infants and adults. AB - OBJECTIVES: Changes in palmar and plantar skin conductance (SC) are due to outgoing bursts in the postganglionic efferent sympathetic cholinergic fibres, which responds to changes in central arousal state. The purpose of this study was designed to develop a software program for analyzing spontaneous and external elicited SC changes for infants and adults. METHODS: The program was designed to calculate the number and mean amplitude of the waves as well as the mean basal level in a given period. Different pre-set values for the minimum amplitude, maximum slope and minimum width of the spontaneous waves were used in the analysis program, and the results were compared with manually counted waves. The program was also used to perform coherent averaging of repeated elicited SC changes. For the mean elicited skin conductance responses, the latency time, response amplitude and recovery time were calculated. The habituation pattern could be calculated semi-automatically by analyzing each response. RESULTS: For SC waves, the minimum amplitude and the maximum slope should be, respectively, 0.02 microsiemens (microS) and 2 microS/s for infants and adults, and the width of the waves should be at least 1 s for adults, and unlimited for infants. The coherent average method was found to be a satisfactory method for revealing whether a subject responded to stimuli, and is recommended, especially for infants. CONCLUSIONS: Spontaneous and stimulated skin conductance are easily analyzed by this software program. PMID- 11018508 TI - Infants' tracking of objects and collections. AB - Recent research suggests that infants' understanding of the physical world is more complex and adult-like than previously believed. One of the most impressive discoveries has been infants' ability to reason about medium-sized, material objects. They are able to individuate objects in a scene, and to enumerate and reason about them. This article reports a series of experiments investigating 8 month-old infants' ability to reason about collections of objects. Experiment 1 shows a sharp contrast between infants' understanding of single objects versus collections. While infants detected the discontinuous ('Magical') disappearance of a single object, they did not detect the Magical Disappearance of a non cohesive pile of objects. Experiments 2-4 found that infants' difficulty remained even when the distinct identity of each object in the collection was emphasized, but could be overcome if infants (a) first saw the individual objects clearly separated from each other prior to their being placed together in a pile, or (b) had prior experience with the objects making up the collection. Our findings suggest that infants' expectations about object behavior are highly specific regarding the entities they are applied to. They do not automatically apply to any and all portions of matter within the visual field. Both the behavior of an entity, and infants' prior experience play roles in determining whether infants will treat that entity as an object. PMID- 11018509 TI - Frequency versus probability formats in statistical word problems. AB - Three experiments examined people's ability to incorporate base rate information when judging posterior probabilities. Specifically, we tested the (Cosmides, L., & Tooby, J. (1996). Are humans good intuitive statisticians after all? Rethinking some conclusions from the literature on judgement under uncertainty. Cognition, 58, 1-73) conclusion that people's reasoning appears to follow Bayesian principles when they are presented with information in a frequency format, but not when information is presented as one case probabilities. First, we found that frequency formats were not generally associated with better performance than probability formats unless they were presented in a manner which facilitated construction of a set inclusion mental model. Second, we demonstrated that the use of frequency information may promote biases in the weighting of information. When participants are asked to express their judgements in frequency rather than probability format, they were more likely to produce the base rate as their answer, ignoring diagnostic evidence. PMID- 11018510 TI - Updating egocentric representations in human navigation. AB - Seven experiments tested whether human navigation depends on enduring representations, or on momentary egocentric representations that are updated as one moves. Human subjects pointed to unseen targets, either while remaining oriented or after they had been disoriented by self-rotation. Disorientation reduced not only the absolute accuracy of pointing to all objects ('heading error') but also the relative accuracy of pointing to different objects ('configuration error'). A single light providing a directional cue reduced both heading and configuration errors if it was present throughout the experiment. If the light was present during learning and test but absent during the disorientation procedure, however, subjects showed low heading errors (indicating that they reoriented by the light) but high configuration errors (indicating that they failed to retrieve an accurate cognitive map of their surroundings). These findings provide evidence that object locations are represented egocentrically. Nevertheless, disorientation had little effect on the coherence of pointing to different room corners, suggesting both (a) that the disorientation effect on representations of object locations is not due to the experimental paradigm and (b) that room geometry is captured by an enduring representation. These findings cast doubt on the view that accurate navigation depends primarily on an enduring, observer-free cognitive map, for humans construct such a representation of extended surfaces but not of objects. Like insects, humans represent the egocentric distances and directions of objects and continuously update these representations as they move. The principal evolutionary advance in animal navigation may concern the number of unseen targets whose egocentric directions and distances can be represented and updated simultaneously, rather than a qualitative shift in navigation toward reliance on an allocentric map. PMID- 11018511 TI - The development of rhythmic attending in auditory sequences: attunement, referent period, focal attending. AB - This paper is divided into three sections. The first section is theoretical; it extends Dynamic Attending Theory (Jones, M. R. Psychological Review 83 (1976) 323; Jones, M. R. Perception and Psychophysics 41(6) (1987) 631; Jones, M. R. Psychomusicology 9(2) (1990) 193; Jones, M. R., & Boltz, M. Psychological Review 96(3) (1989) 459) to developmental questions concerning tempo and time hierarchies. Generally Dynamic Attending Theory proposes that, when listening to a complex auditory sequence, listeners spontaneously focus on events occurring at an intermediate rate (the referent level), and they then may shift attention to events occurring over longer or shorter time spans, that is at lower (faster) or higher (slower) hierarchical levels (focal attending). The second section of the paper is experimental. It examines maturational changes of three dynamic attending activities involving referent period and level, attunement, and focal attending. Tasks involve both motor tapping (including spontaneous motor tempo and synchronization with simple sequences and music) and tempo discrimination. We compare performances by 4-, 6-, 8-, and 10-year-old children and adults, with or without musical training. Results indicate three changes with increased age and musical training: (1) a slowing of the mean spontaneous tapping rate (a reflection of the referent period) and mean synchronization rate (a reflection of the referent level), (2) enhanced ability to synchronize tapping and discriminate tempo (improved attunement), and (3) an enlarged range of tapping rates towards slower rates and higher hierarchical levels (improved focal attending). A final section considers results in light of the theory proposed here. It is suggested that growth trends can be expressed in terms of listeners' engagement of slower attending oscillators with age and experience, accompanied by the passage from the initial use of a single oscillator towards the coupling of multiple oscillators. PMID- 11018512 TI - The separate but related origins of the recency effect and the modality effect in free recall. AB - The recency effect found in free recall can be accounted for almost entirely in terms of the recall of ordered sequences of items. It is such sequences, presented at the end of the stimulus list but recalled at the very beginning of the response protocol, which produce a recency effect. Such sequences are recalled at the beginning of the response protocol equally often following auditory and visual presentation. These same stimulus sequences are also frequently recalled other than initially in the response protocol following auditory presentation. However, such responses are rarely found following visual presentation. The modality effect in free recall, the advantage of auditory over visual presentation, can be substantially accounted for in these terms. Theoretical and procedural implications of these data are discussed. PMID- 11018513 TI - Ankyrins. AB - This review is focused on ankyrin which is a protein linker between the integral membrane proteins and spectrin-based cytoskeleton. Structure and distribution of different ankyrin isoforms that are products of alternative-spliced genes are described. Interaction of ankyrins with various membranes is considered. Special attention is paid to ankyrin participation in signal transduction and in assembly of integral membrane proteins in specialized membrane domains. PMID- 11018514 TI - Mitochondria: execution central. AB - Mitochondria play an essential function in eukaryotic life and death. They also play a central role in apoptosis regulation, reflected by the convergence of Bcl 2 family members on the mitochondrial outer membrane, and the presence of 'death factors' in the intermembrane space. Mitochondrial structure and function must be taken into consideration when evaluating mechanisms for cytochrome c release. The core machinery for caspase activation is conserved from Caenorhabditis elegans to man, and we consider parallels in the role of mitochondria in this process. PMID- 11018515 TI - Conservation of components of the dystrophin complex in Drosophila. AB - Defects in the dystrophin complex (DC) underlie several human genetic disorders, but our dissection of its function is complicated by potential redundancy of the multiple vertebrate isoforms of most DC components. We here complete our previous description of Drosophila dystrophin, and show that the fly retains all essential components of the DC, but with substantially less diversity. Seventeen known human components (three dystrophin-related proteins, two dystrobrevins, five sarcoglycans, five syntrophins, one dystroglycan and one sarcospan) appear to be reduced to eight in Drosophila (one, one, three, two, one and none, respectively). The simplicity of this system recommends it as a model for its human counterpart. PMID- 11018516 TI - CEO1, a new protein from Arabidopsis thaliana, protects yeast against oxidative damage. AB - The Saccharomyces cerevisiae strain WYT, deficient in the YAP1 transcription factor, was used in a molecular screen to identify genes from Arabidopsis thaliana that could overcome the oxidative stress-sensitive phenotype of these yeast cells. A cDNA named CEO1 increased the tolerance to oxidative damage caused by tert-butylhydroperoxide of both the Yap1(-) mutant and the wild-type yeast. Additionally, in Yap1(-) yeast, CEO1 also induced cross-tolerance to oxidative damage caused by hydrogen peroxide and diamide. CEO1 was assigned as being part of a small gene family that, until now, is exclusively restricted to plants. In Arabidopsis, CEO1 was produced in all organs, especially in roots and stems. By using the yeast two-hybrid system, proteins that specifically interact with CEO1 in yeast were identified, and putative DNA-binding proteins were consistently recovered. PMID- 11018517 TI - Expression of doubly labeled Saccharomyces cerevisiae iso-1 ferricytochrome c and (1)H, (13)C and (15)N chemical shift assignments by multidimensional NMR. AB - We have expressed [U-(13)C,(15)N]-labeled Saccharomyces cerevisiae iso-1 cytochrome c C102T;K72A in Escherichia coli with a yield of 11 mg/l of growth medium. Nuclear magnetic resonance (NMR) studies were conducted on the Fe(3+) form of the protein. We report herein chemical shift assignments for amide (1)H and (15)N, (13)C(omicron), (13)C(alpha), (13)C(beta), (1)H(alpha) and (1)H(beta) resonances based upon a series of three-dimensional NMR experiments: HNCA, HN(CO)CA, HNCO, HN(CA)CO, HNCACB, HCA(CO)N, HCCH-TOCSY and HBHA(CBCA)NH. An investigation of the chemical shifts of the threonine residues was also made by using density functional theory in order to help solve discrepancies between (15)N chemical shift assignments reported in this study and those reported previously. PMID- 11018518 TI - Duplication of genes encoding non-clathrin coat protein gamma-COP in vertebrate, insect and plant evolution. AB - Coatomer is a major component of COPI vesicles and consists of seven subunits. The gamma-COP subunit of the coatomer is believed to mediate the binding to the cytoplasmic dilysine motifs of membrane proteins. We characterized cDNAs for Copg genes encoding gamma-COP from mouse, zebrafish, Drosophila melanogaster and Bombyx mori. Two copies of Copg genes are present in vertebrates and in B. mori. Phylogenetic analysis revealed that two paralogous genes had been derived from a single ancestral gene by duplication independently in vertebrates and in B. mori. Mouse Copg1 showed ubiquitous expression with the highest level in testis. Zebrafish copg2 was biallelically expressed in hybrid larvae in contrast to its mammalian ortholog expressed in a parent-of-origin-specific manner. A phylogenetic analysis with partial plant cDNA sequences suggested that copg gene was also duplicated in the grass family (Poaceae). PMID- 11018519 TI - Maternal cold inducible RNA binding protein is required for embryonic kidney formation in Xenopus laevis. AB - We cloned a major isoform of Xenopus homologue of cold inducible RNA binding protein (CIRP), XCIRP-1. XCIRP-1 was neither cold inducible nor essential for cell division during early embryonic development. Suppression of XCIRP-1 dose dependently produced tailbuds with deformations of the brain and internal organs. The defects were XCIRP-1 specific as they could be rescued by sense transcript. Suppression of XCIRP-1 also disrupted the morphogenetic migration of the C3 blastomeres (lineaged to become the embryonic kidney, the pronephros). In animal cap explants, depletion of XCIRP-1 inhibited activin/retinoic acid induced expressions of pronephros related Xlim-1 and WT1 genes. These results suggest that XCIRP-1 is required for the specification and morphogenetic lineage migration of the pronephros. PMID- 11018520 TI - MSK1 is required for CREB phosphorylation in response to mitogens in mouse embryonic stem cells. AB - Mouse embryonic stem (ES) cells homozygous for disruption of the MSK1 gene had no detectable MSK1 activity. However, their activators (extracellular signal related kinase (ERK)1/ERK2) were stimulated normally in mitogen- and stress-activated protein kinase (MSK)1-/- and wild type cells in response to tetradecanoylphorbol acetate (TPA) and epidermal growth factor (EGF). TPA and EGF induced the phosphorylation of cyclic AMP-responsive element binding protein (CREB) at Ser 133 and ATF1 at Ser-63 in wild type cells and this was abolished by inhibition of the mitogen-activated protein kinase cascade. In contrast, the TPA- and EGF induced phosphorylation of CREB/ATF1 was barely detectable in MSK1-/- cells. However, basal and forskolin-induced phosphorylation was similar, indicating that the MSK1 'knockout' did not prevent CREB phosphorylation by cyclic AMP-dependent protein kinase. Thus MSK1 is required for CREB and ATF1 phosphorylation after mitogenic stimulation of ES cells. PMID- 11018521 TI - Analysis of dipeptidyl peptidase IV gene regulation in transgenic mice: DNA elements sufficient for promoter activity in the kidney, but not the intestine, reside on the proximal portion of the gene 5'-flanking region. AB - The dipeptidyl peptidase IV (DPPIV) gene encodes a brush border membrane exopeptidase that is expressed in a tissue-restricted fashion. To examine the regulation of DPPIV transcription in various tissues in vivo, we examined the expression of DPPIV 5'-flanking region (promoter)-human growth hormone reporter constructs in transgenic mice. These mice exhibited cell-type specific reporter expression in kidney. Surprisingly, however, only very low to non-detectable levels of reporter were found in small intestine. These results indicate that DNA elements sufficient for DPPIV expression in kidney, but not intestine, reside in the 5'-flanking region of the gene. PMID- 11018522 TI - The direct association of the multiple PDZ domain containing proteins (MUPP-1) with the human c-Kit C-terminus is regulated by tyrosine kinase activity. AB - We have identified the multiple PDZ domain containing protein (MUPP-1 or MPDZ) as a novel binding partner of the human c-Kit. c-Kit binds specifically to the 10th PDZ domain of MUPP-1 via its C-terminal sequence. Furthermore, a kinase negative mutant receptor interacted more strongly with MUPP-1 than the wild-type c-Kit. Strikingly, a constitutively activated c-Kit (D816V-Kit) did not bind to MUPP-1, although this oncogenic form retains the PDZ binding motif 'HDDV' at the C terminal end. Deletion of V967 of c-Kit abolished binding to MUPP-1 and drastically reduced its tyrosine kinase activity, suggesting that the structure of the C-terminal tail of c-Kit influences its enzymatic activity. PMID- 11018523 TI - Investigation of the molecular assembly of beta-cell K(ATP) channels. AB - We have investigated the protein interactions involved in the assembly of pancreatic beta-cell ATP-sensitive potassium channels. The channels are a heterooligomeric complex of pore-forming Kir6.2 subunits and sulfonylurea receptor (SUR1) subunits. SUR1 belongs to the ATP binding cassette (ABC) family of proteins and has two nucleotide binding domains (NBD1 and NBD2) and 17 putative transmembrane (TM) sequences. Previously we showed that co-expression in a baculovirus expression system of two parts of SUR1 divided at Pro1042 between TM12 and 13 leads to restoration of glibenclamide binding activity, whereas expression of either individual N- or C-terminal domain alone gave no glibenclamide binding activity [M.V. Mikhailov and S.J.H. Ashcroft (2000) J. Biol. Chem. 275, 3360-3364]. Here we show that the two half-molecules formed by division of SUR1 between NBD1 and TM12 or between TM13 and 14 also self-assemble to give glibenclamide binding activity. However, deletion of NBD1 from the N-part of SUR1 abolished SUR1 assembly, indicating a critical role for NBD1 in SUR1 assembly. We found that differences in glibenclamide binding activity obtained after co-expression of different half-molecules are attributable to different amounts of binding sites, but the binding affinities remained nearly the same. Simultaneous expression of Kir6.2 resulted in enhanced glibenclamide binding activity only when the N-half of SUR1 included TM12. We conclude that TM12 and 13 are not essential for SUR1 assembly whereas TM12 takes part in SUR1 Kir6.2 interaction. This interaction is specific for Kir 6.2 because no enhancement of glibenclamide binding was observed when half-molecules were expressed together with Kir4.1. We propose a model of K(ATP) channel organisation based on these data. PMID- 11018524 TI - Neuronal nitric oxide synthase localizes through multiple structural motifs to the sarcolemma in mouse myotubes. AB - In skeletal muscle, neuronal nitric oxide synthase is localized at the sarcolemma in association with the dystrophin glycoprotein complex (DGC). The nNOS N terminal 231 amino acids comprise a PDZ domain (residues 1-100) and a beta hairpin finger loop (residues 101-130) which binds alpha-syntrophin located in the DGC. Endogenous nNOS and GFP-tagged nNOS localize to the sarcolemma in mouse C2C12 myotubes. Expression of GFP-tagged nNOS domains in C2C12 myotubes reveals that the PDZ domain and the beta-hairpin finger loop of nNOS are independently capable of localizing to the sarcolemma of C2C12 myotubes. Binding studies indicate that alpha-syntrophin binds only to the beta-hairpin finger loop and not the PDZ domain of nNOS. nNOS may bind to proteins in addition to alpha-syntrophin at muscle sarcolemma. PMID- 11018525 TI - New molecular aspects of regulation of mitochondrial activity by fenofibrate and fasting. AB - Fenofibrate and fasting are known to regulate several genes involved in lipid metabolism in a similar way. In this study measuring several mitochondrial enzyme activities, we demonstrate that, in contrast to citrate synthase and complex II, cytochrome c oxidase (COX) is a specific target of these two treatments. In mouse liver organelles, Western blot experiments indicated that mitochondrial levels of p43, a mitochondrial T3 receptor, and mitochondrial peroxisome proliferator activated receptor (mt-PPAR), previously described as a dimeric partner of p43 in the organelle, are increased by both fenofibrate and fasting. In addition, in PPAR alpha-deficient mice, this influence was abolished for mt-PPAR but not for p43, whereas the increase in COX activity was not altered. These data indicate that: (1) PPAR alpha is involved in specific regulation of mt-PPAR expression by both treatments; (2) fenofibrate and fasting regulate the mitochondrial levels of p43 and thus affect the efficiency of the direct T3 mitochondrial pathway. PMID- 11018526 TI - Overexpression of a dominant negative CREB protein in HT-1080 cells selectively disrupts plasminogen activator inhibitor type 2 but not tissue-type plasminogen activator gene expression. AB - The tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor type 2 (PAI-2) genes are differentially regulated by 12-phorbol 13-myristate acetate (PMA) in HT-1080 fibrosarcoma cells. PMA transcriptionally down-regulates the t-PA gene in HT-1080 cells, while the PAI-2 gene is simultaneously induced by this agonist. The t-PA and PAI-2 gene promoters harbour a cAMP-response element (CRE) which influences the expression of both genes. We have compared the binding activity of nuclear factors that recognise these CRE sites. We show that CREB (CRE binding protein) recognises each CRE and that the degree of constitutive Ser119-phosphorylated t-PA CRE-bound CREB was greater than for PAI-2 CRE bound CREB. Stable transfection of HT-1080 cells with a plasmid containing a CREB that could not be phosphorylated on Ser119 (pCI-CREB(ala119)) did not influence PMA mediated suppression of t-PA mRNA, but markedly impaired PMA-mediated induction of PAI-2 mRNA. Our results demonstrate that the Ser119 residue of CREB plays a crucial role in PMA-mediated induction of PAI-2 gene expression, whereas PMA mediated suppression of t-PA in HT-1080 cells requires a different process. PMID- 11018527 TI - Biophysical characterization of SipA, an actin-binding protein from Salmonella enterica. AB - An essential step in the pathogenesis of Salmonella enterica infections is bacterial entry into non-phagocytic cells of the intestinal epithelium. Proteins injected by Salmonella into host cells stimulate cellular responses that lead to extensive actin cytoskeleton reorganization and subsequent bacterial uptake. One of these proteins, SipA, modulates actin dynamics by directly binding to F-actin. We have biophysically characterized a C-terminal fragment, SipA(446-684), which has previously been shown to retain activity. Our results show that SipA(446-684) exhibits an elongated shape with a predominantly helical conformation and predict the existence of a coiled-coil domain. We suggest that the protein is able to span two adjacent actin monomers in a filament and propose a model that is consistent with the observed effects of SipA(446-684) on actin dynamics and F actin stability and morphology. PMID- 11018528 TI - Sphingosylphosphorylcholine induces Ca(2+)-sensitization of vascular smooth muscle contraction: possible involvement of rho-kinase. AB - Sphingosylphosphorylcholine (SPC), a sphingolipid, concentration-dependently (1 50 microM) induced contraction and slight elevation of the cytosolic Ca(2+) concentration ([Ca(2+)](i)) in smooth muscle of the pig coronary artery, the result being a marked increase in the force/[Ca(2+)](i) ratio. In alpha-toxin- or beta-escin-permeabilized, but not Triton X-100-permeabilized, vascular strips, SPC induced contraction at constant [Ca(2+)](i) (pCa 6.3) in the absence of GTP, whereas a G-protein-coupled receptor agonist, histamine, required the presence of GTP to induce the contraction. The Rho-kinase blocker, Y-27632 (10 microM) abolished the SPC-induced Ca(2+)-sensitization, without affecting the Ca(2+) induced contraction. These results suggest that SPC induces Ca(2+)-sensitization of force in vascular smooth muscle, presumably through the activation of Rho kinase (or a related kinase). PMID- 11018529 TI - Epoxysuccinyl peptide-derived affinity labels for cathepsin B. AB - Extracellular cysteine proteases, in particular cathepsin B, have been implicated in a variety of pathological processes. Selectively targeting labels of this enzyme are important tools to gain more detailed understanding of its specific roles. Starting from our recently developed irreversible epoxysuccinyl-based inhibitor (R-Gly-Gly-Leu-(2S,3S)-tEps-Leu-Pro-OH, R=OMe), we have synthesized two affinity labels, R=NH-(CH(2))(6)-NH-rhodamine B and R=NH-(CH(2))(6)-NH-biotin. Using MCF-7 cells, the labeled inhibitors were shown to be virtually non-cell permeant. Moreover, affinity blot analysis with the biotinylated inhibitor allowed a highly sensitive and selective non-radioactive detection of active cathepsin B. PMID- 11018530 TI - Permissive role of protein kinase C alpha but not protein kinase C delta in sphingosine 1-phosphate-induced Rho A activation in C2C12 myoblasts. AB - Rho GTPases participate in various important signaling pathways and have been implicated in myogenic differentiation. Here the first evidence is provided that in C2C12 myoblasts sphingosine 1-phosphate (SPP) rapidly and transiently induced membrane association of Rho A in a pertussis toxin-insensitive manner. The bioactive lipid preferentially relocalized the GTPase to Golgi-enriched membrane. Translocation of Rho A was abolished by inhibition or down-regulation of protein kinase C (PKC). Notably, treatment with Go6976, an inhibitor of conventional PKCs, which selectively blocked PKC alpha in these cells, prevented SPP-induced Rho A translocation. Conversely rottlerin, a selective inhibitor of PKC delta, was without effect, demonstrating that SPP signaling to Rho A involves PKC alpha but not PKC delta activation. This novel functional relationship between the two proteins may have a role in SPP-mediated regulation of downstream effectors. PMID- 11018531 TI - Characterization of Brassica S-haplotypes lacking S-locus glycoprotein. AB - Self-incompatibility (SI) in Brassica is regulated by a single multi-allelic locus, S, which contains highly polymorphic stigma-expressed genes, SLG and SRK. While SRK is shown to be the determinant of female SI specificity, SLG is thought to assist the function of SRK. Here we report that the SLG genes of self incompatible S(18) and S(60) homozygotes of Brassica oleracea have an in-frame stop codon and a 23 bp deletion resulting in a frame-shift, respectively. The finding that these SLG genes do not encode functional SLG proteins suggests that SLG is not essential for SI. The possible role of SLG in SI was discussed. PMID- 11018532 TI - Interactions of bile salt micelles and colipase studied through intermolecular nOes. AB - Colipase is a small protein (10 kDa), which acts as a protein cofactor for the pancreatic lipase. Various models of the activated ternary complex (lipase colipase-bile salt micelles) have been proposed using detergent micelles, but no structural information has been established with bile salt micelles. We have investigated the organization of sodium taurodeoxycholate (NaTDC) micelles and their interactions with pig and horse colipases by homonuclear nuclear magnetic resonance (NMR) spectroscopy. The NMR data supply evidence that the folding of horse colipase is similar to that already described for pig colipase. Intermolecular nuclear Overhauser effects have shown that two conserved aromatic residues interact with NaTDC micelles. PMID- 11018533 TI - Isolation and characterization of a higher plant ADP-glucose pyrophosphorylase small subunit homotetramer. AB - ADP-glucose pyrophosphorylase (AGPase) is the allosterically regulated gateway for carbon entry into transient and storage starch in plants as well as glycogen in bacteria. This enzyme plays a key role in the modulation of photosynthetic efficiency in source tissues and directly determines the level of storage starch in sink tissues, thus influencing overall crop yield potential. AGPase is a tetrameric enzyme; in higher plants it consists of two regulatory large subunits (LS) and two catalytic small subunits (SS), while in cyanobacteria and prokaryotes the enzyme is homotetrameric. The potato SS gene in pML10 was mutated by hydroxylamine and mutants were screened for elevated homotetrameric activity by iodine vapor staining. This search strategy led to the isolation of SS mutants (SUP-1, TG-15) that had pyrophosphorylase activity in the absence of the LS. TG 15 has a leucine to phenylalanine change at position 48 (L(48)F) that corresponds to a phenylalanine residue at the analogous position in the Escherichia coli homotetrameric AGPase as well as a valine to isoleucine change at position 59 (V(59)I). TG-15 was partially purified and kinetic analysis revealed substrate and effector affinities equal to wild type heterotetrameric enzyme with the exception of ATP binding. PMID- 11018534 TI - Structure-based sequence alignment for the beta-trefoil subdomain of the clostridial neurotoxin family provides residue level information about the putative ganglioside binding site. AB - Clostridial neurotoxins embrace a family of extremely potent toxins comprised of tetanus toxin (TeNT) and seven different serotypes of botulinum toxin (BoNT/A-G). The beta-trefoil subdomain of the C-terminal part of the heavy chain (H(C)), responsible for ganglioside binding, is the most divergent region in clostridial neurotoxins with sequence identity as low as 15%. We re-examined the alignment between family sequences within this subdomain, since in this region all alignments published to date show obvious inconsistencies with the beta-trefoil fold. The final alignment was obtained by considering the general constraints imposed by this fold, and homology modeling studies based on the TeNT structure. Recently solved structures of BoNT/A confirm the validity of this structure-based approach. Taking into account biochemical data and crystal structures of TeNT and BoNT/A, we also re-examined the location of the putative ganglioside binding site and, using the new alignment, characterized this site in other BoNT serotypes. PMID- 11018535 TI - Nuclear localization of a hypoxia-inducible novel non-symbiotic hemoglobin in cultured alfalfa cells. AB - We have isolated a 483-bp-long full-length cDNA clone encoding a non-symbiotic hemoglobin called Mhb1, the first one found in alfalfa. This non-symbiotic hemoglobin is a single copy gene localized in linkage group 4 in diploid Medicago genome. The Mhb1 mRNA was found only in the roots of alfalfa plants. The Mhb1 gene was inducible by hypoxia and showed no induction by cold stress treatment. The Mhb1 transcript level increased at the G2/M boundary in a synchronized alfalfa cell suspension culture. The majority of Mhb1 protein was shown to be localized in the nucleus and smaller amounts were detected in the cytoplasm. A potential link to the nitric oxide signalling pathway is also discussed. PMID- 11018536 TI - Intracellular Ca(2+) channel immunoreactivity in neuroendocrine axon terminals. AB - The concentration of neuroendocrine terminals in the neurohypophysis facilitates the identification and localization of Ca(2+) channel subtypes near neuroendocrine release sites. Immunoblots of rat neurohypophysial tissue identified the alpha(1)1.3, alpha(1)2.1, alpha(1)2.2, and alpha(1)2.3 Ca(2+) channel subunits. Immunofluorescence staining of axon terminal plasma membranes was weak, suggesting that Ca(2+) channels are dispersed. This contrasts with the highly punctate alpha(1)2.2 immunoreactivity in bovine chromaffin cells; the neurohypophysial terminals may therefore lack the specialized release zones found in those cells. Immunofluorescence and immunogold labeling identify dense core granule-like structures in the terminal cytoplasm containing multiple Ca(2+) channel types. Ca(2+) channels in internal membranes may play an important role in channel targeting and distribution in neuroendocrine cells. PMID- 11018537 TI - The Yersinia Ser/Thr protein kinase YpkA/YopO directly interacts with the small GTPases RhoA and Rac-1. AB - Pathogenic bacteria of the genus Yersinia counteract host defense by interfering with eukaryotic signal transduction pathways. YpkA of Yersinia pseudotuberculosis shares significant homology with eukaryotic Ser/Thr protein kinases, is translocated into the host cell and has been shown to be an essential virulence factor in a mouse infection model. In this study, we identify the small GTPases RhoA and Rac-1 as eukaryotic binding partners of YpkA and its homolog YopO of Yersinia enterocolitica. We demonstrate that the interaction is independent of phosphorylation of YpkA and nucleotide loading state of the GTPases. The interaction with RhoA and Rac-1 might provide an important clue to how YpkA interferes with eukaryotic signaling on a molecular level. PMID- 11018538 TI - Structure of the 5th transmembrane segment of the Na,K-ATPase alpha subunit: a cysteine-scanning mutagenesis study. AB - To study the structure of the pathway of cations across the Na, K-ATPase, we applied the substituted cysteine accessibility method to the putative 5th transmembrane segment of the alpha subunit of the Na,K-ATPase of the toad Bufo marinus. Only the most extracellular amino acid position (A(796)) was accessible from the extracellular side in the native Na,K-pump. After treatment with palytoxin, six other positions (Y(778), L(780), S(782), P(785), E(786) and L(791)), distributed along the whole length of the segment, became readily accessible to a small-size methanethiosulfonate compound (2-aminoethyl methanethiosulfonate). The accessible residues are not located on the same side of an alpha-helical model but the pattern of reactivity would rather suggest a beta-sheet structure for the inner half of the putative transmembrane segment. These results demonstrate the contribution of the 5th transmembrane segment to the palytoxin-induced channel and indicate which amino acid positions are exposed to the pore of this channel. PMID- 11018539 TI - Activation of vanadium nitrogenase expression in Azotobacter vinelandii DJ54 revertant in the presence of molybdenum. AB - Azotobacter vinelandii carries three different and genetically distinct nitrogenase systems on its chromosome. Expression of all three nitrogenases is repressed by high concentrations of fixed nitrogen. Expression of individual nitrogenase systems is under the control of specific metal availability. We have isolated a novel type of A. vinelandii DJ54 revertant, designated A. vinelandii BG54, which carries a defined deletion in the nifH gene and is capable of diazotrophic growth in the presence of molybdenum. Inactivation of nifDK has no effect on growth of this mutant strain in nitrogen-free medium suggesting that products of the nif system are not involved in supporting diazotrophic growth of A. vinelandii BG54. Similar to the wild type, A. vinelandii BG54 is also sensitive to 1 mM tungsten. Tn5-B21 mutagenesis to inactivate the genes specific to individual systems revealed that the structural genes for vnf nitrogenase are required for diazotrophic growth of A. vinelandii BG54. Analysis of promoter activity of different nif systems revealed that the vnf promoter is activated in A. vinelandii BG54 in the presence of molybdenum. Based on these data we conclude that A. vinelandii BG54 strain utilizes vnf nitrogenase proteins to support its diazotrophic growth. PMID- 11018540 TI - Binding of a de novo designed peptide to specific glycosaminoglycans. AB - The binding of glycosaminoglycans to a synthetic peptide (SKAQKAQAKQAKQAQKAQKAQAKQAKQW-CONH(2)), consisting of a hybrid consensus heparin binding sequence, is studied using circular dichroism, fluorescence anisotropy and nuclear magnetic resonance techniques. The results unveil certain novel features, most importantly, the peptide binds preferentially to iduronic acid containing glycosaminoglycans and the dissociation constant for the peptide heparin complex was found to be 30 nM. Interestingly, higher order intermolecular association(s)/aggregation was not observed, especially at saturating concentrations of the ligand. The helical structure of the peptide backbone, induced upon binding to a particular glycosaminoglycan is directly related to their binding affinity. In our opinion, studies on such unconventional hybrid peptide sequences containing low density basic amino acid residues would lead to the design of sequence specific glycosaminoglycan binding peptides. PMID- 11018541 TI - Observation of RecA protein monomer by small angle X-ray scattering with synchrotron radiation. AB - RecA protein is capable of forming homo-oligomers in solution. The oligomeric and monomeric states of Thermus thermophilus RecA protein were studied by small angle X-ray scattering, a direct method used to measure the overall dimensions of a macromolecule. In the presence of 3 M urea or 0.2 M lithium perchlorate, RecA dissociates from higher oligomeric states to form a hexamer with a radius of gyration (R(g)) of 52 A. The value of R(g) decreased to 36 A at a higher lithium perchlorate concentration (1.0 M). The zero angle intensity, I(0), was consistent with the identification of the former state as a hexamer and the latter as a monomer. PMID- 11018542 TI - Functional domains of the ATPase inhibitor protein from bovine heart mitochondria. AB - A study is presented of the activity and temperature dependence of the ATPase inhibitor protein (IF(1)) from bovine heart mitochondria and of synthetic partial IF(1) peptides. The results show that the IF(1)-(42-58) peptide is the most potent inhibitory domain of IF(1). PMID- 11018543 TI - Dextrans for targeted and sustained delivery of therapeutic and imaging agents. AB - Dextrans are glucose polymers which have been used for more than 50 years as plasma volume expanders. Recently, however, dextrans have been investigated for delivery of drugs, proteins/enzymes, and imaging agents. These highly water soluble polymers are available commercially as different molecular weights (M(W)) with a relatively narrow M(W) distribution. Additionally, dextrans contain a large number of hydroxyl groups which can be easily conjugated to drugs and proteins by either direct attachment or through a linker. In terms of pharmacokinetics, the intact polymer is not absorbed to a significant degree after oral administration. Therefore, most of the applications of dextrans as macromolecular carriers are through injectable routes. However, a few studies have reported the potential of dextrans for site (colon)-specific delivery of drugs via the oral route. After the systemic administration, the pharmacokinetics of the conjugates of dextran with therapeutic/imaging agents are significantly affected by the kinetics of the dextran carrier. Animal and human studies have shown that both the distribution and elimination of dextrans are dependent on the M(W) and charge of these polymers. Pharmacodynamically, conjugation with dextrans has resulted in prolongation of the effect, alteration of toxicity profile, and a reduction in the immunogenicity of drugs and/or proteins. A substantial number of studies on dextran conjugates of therapeutic/imaging agents have reported favorable alteration of pharmacokinetics and pharmacodynamics of these agents. However, most of these studies have been carried out in animals, with only a few being extended to humans. Future studies should concentrate on barriers for the clinical use of dextrans as macromolecular carriers for delivery of drugs, proteins, and imaging agents. PMID- 11018544 TI - Synthesis and HPLC analysis of enzymatically cleavable linker consisting of poly(ethylene glycol) and dipeptide for the development of immunoconjugate. AB - A model compound of anti-tumor agent, segment B of duocarmycin derivative DU-86, was conjugated to tumor-specific antibody via a cleavable linker consisting of poly(ethylene glycol) (PEG) and dipeptide, L-alanyl-L-valine (Ala-Val), to confirm the feasibility of the linker for application to immunoconjugate. The release of segment B from the linker was evaluated by HPLC analysis. When segment B was derivatized to have an amino residue and then linked to PEG through a dipeptide, segment B was cleaved at the peptide bond by a particular enzyme, thermolysin (EC 3.4.24.4), but not by plasmin (EC 3.4.2 1.7.), indicating that certain protease specifically expressed at the tumor site would be capable of peptide-specific digestion and release of anti-tumor agent since a thermolysin like enzyme has been reported to be expressed at many tumor cells. Furthermore, the results showing that cell extract from G361 human melanoma had an ability to digest the linker peptide while the linker was stable in normal human serum suggested the tumor-specific activation of the conjugated agent. Segment B was conjugated via the linker to murine monoclonal antibody KM641 reactive to GD3 ganglioside to form immunoconjugate and the quantitative release of segment B under the treatment with the enzyme was also confirmed. These results indicate the possibility of double targeting based on both the recognition ability of tumor specific antibody and tumor specific activation of the anti-tumor agents to enhance tumor treatment efficacy and to decrease unwanted side effects. PMID- 11018545 TI - Factors affecting acoustically triggered release of drugs from polymeric micelles. AB - A custom ultrasonic exposure chamber with real-time fluorescence detection was used to measure acoustically-triggered drug release from Pluronic P-105 micelles under continuous wave (CW) or pulsed ultrasound in the frequency range of 20 to 90 kHz. The measurements were based on the decrease in fluorescence intensity when drug was transferred from the micelle core to the aqueous environment. Two fluorescent drugs were used: doxorubicin (DOX) and its paramagnetic analogue, ruboxyl (Rb). Pluronic P-105 at various concentrations in aqueous solutions was used as a micelle-forming polymer. Drug release was most efficient at 20-kHz ultrasound and dropped with increasing ultrasonic frequency despite much higher power densities. These data suggest an important role of transient cavitation in drug release. The release of DOX was higher than that of Rb due to stronger interaction and deeper insertion of Rb into the core of the micelles. Drug release was higher at lower Pluronic concentrations, which presumably resulted from higher local drug concentrations in the core of Pluronic micelles when the number of micelles was low. At constant frequency, drug release increased with increasing power density. At constant power density and for pulse duration longer than 0.1 s, peak release under pulsed ultrasound was the same as stationary release under CW ultrasound. Released drug was quickly re-encapsulated between the pulses of ultrasound, which suggests that upon leaving the sonicated volume, the non-extravasated and non-internalized drug would circulate in the encapsulated form, thus preventing unwanted drug interactions with normal tissues. PMID- 11018546 TI - Adjuvancy enhancement of muramyl dipeptide by modulating its release from a physicochemically modified matrix of ovalbumin microspheres. I. In vitro characterization. AB - The weak immunogenicity of subunit vaccines has necessitated research into the development of novel adjuvants and methods to enhance the adjuvancy associated with vaccine delivery systems. The purpose of the present study was to modulate the release of muramyl dipeptide (MDP) from a physicochemically modified matrix of ovalbumin microspheres (OVA-MSs). A two-component MS vaccine delivery system was fabricated, which utilized OVA as the antigen and delivery matrix, and MDP as the adjuvant. The MSs were prepared from OVA using a water/oil emulsion method, followed by suspension cross-linking using glutaraldehyde. The MS matrix was modified with respect to the degree of cross-linking by varying the concentration of glutaraldehyde and matrix density, a function of disulfide-bond formation. The modifications in the MS matrix were characterized using SDS-PAGE, scanning electron microscopy, differential scanning calorimetry, and thin layer wicking (TLW). The in vitro release of MDP and OVA from the various preparations of OVA MSs exhibited triphasic and biphasic profiles, respectively. The degree of cross linking and the matrix density were found to be significant physicochemical parameters that affected the release profiles of MDP and OVA through two mechanisms: controlled surface erosion and bulk degradation of the MSs. PMID- 11018547 TI - Adjuvancy enhancement of muramyl dipeptide by modulating its release from a physicochemically modified matrix of ovalbumin microspheres. II. In vivo investigation. AB - In the present study, sustaining the release of adjuvants was investigated using microspheres as a means to increase the immune response (i.e. efficacy) and, ultimately, to reduce adverse effects to vaccine components. To date, most attempts have focused on sustaining the release of antigens. The utility of currently used vaccine adjuvants may be improved by sustaining their release. The development, modification and characterization of a two-component microsphere vaccine delivery system was demonstrated in our previous report [Puri et al., J. Control. Release (2000) in press]. Briefly, ovalbumin (OVA) was utilized as the model antigen (Ag) and delivery matrix and MDP or threonyl-MDP served as the model adjuvants. The release pattern of MDP was modulated from a physicochemically modified matrix of OVA microspheres (OVA-MSs). The purpose of the present study was to evaluate the adjuvancy of MDP in mice by modulating its release from OVA-MSs. Mice were immunized intradermally (i.d.) with various preparations of OVA-MSs, using a single-shot-immunization technique. Positive and negative control preparations were evaluated as well. An inverse relationship was observed between the in vitro release rate of MDP and the in vivo OVA-specific IgG antibody (Ab) immune response in mice. These results demonstrated that modulating the release pattern of MDP or threonyl-MDP enhanced their adjuvant effect. In conclusion, the current results demonstrate that the sustained and controlled release of adjuvants is extremely important for inducing a high level and prolonged period of immunostimulation while potentially minimizing therapy limiting adverse effects. PMID- 11018548 TI - Effect of preparation temperature on the characteristics and release profiles of PLGA microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method. AB - This study describes the influence of preparation temperature on the various characteristics and release profiles of poly(DL-lactide-co-glycolide) (PLGA) microspheres. The bovine serum albumin (BSA)-loaded microspheres were prepared using the water-in-oil-in-water (w/o/w) technique with poly(vinyl alcohol) as surfactant in the external aqueous phase. We have varied the preparation temperature to observe its effect on microsphere characteristics such as the microsphere shrinking rate during formation, particle size, density, surface and internal morphology, BSA encapsulation efficiency, BSA initial release, microsphere degradation and BSA in vitro release behaviour. During fabrication, a low preparation temperature of 5 degrees C gives the fastest initial but the slowest overall shrinking rate. Microspheres formed at high temperatures of 38 degrees C and 42 degrees C on the other hand have the lowest initial yet the highest overall shrinking rate. Subsequently, microsphere mean size increases and the particle size distribution widens with increase in the preparation temperature. Although all the microspheres have a porous surface as well as internal structure, microspheres fabricated at high temperatures have a uniform internal pore distribution and a very thin dense skin layer, while microspheres fabricated at lower temperatures have a thicker but porous skin layer and bigger pores in the middle of the sphere. Microspheres formed at 33 degrees C are found to give the highest initial burst release. In terms of in vitro release, microspheres fabricated at low temperatures (5 degrees C, 15 degrees C and 22 degrees C) exhibit similar, steady rates. Microspheres formed at higher temperatures however give very low release rates after their initial release. The results obtained suggest that preparation temperature significantly affects microsphere formation, resulting in their structural and protein release profile differences. These differences ultimately work together to affect the initial release and overall release patterns of the microspheres. PMID- 11018550 TI - Theoretical analysis of inward hemispheric release above and below drug solubility. AB - A detailed analysis of inward diffusional drug release from devices with hemispheric and related geometries is presented. When drug is loaded below its solubility, an infinite series describes drug concentration profiles and release kinetics, with an excellent approximation resulting when only one term of this series is retained. A connection between this geometric setting and diffusion in constricted porous domains is pointed out, as is the utility of mean first passage times and mean residence times derived for this model. For the case of drug loaded above its solubility, the pseudosteady state (PSS) approximation of Bechard and McMullen [J. Pharm. Sci. 77 (1988) 222] is compared against numerical results calculated for the full model in which the PSS assumption is removed. A close match is observed. Asymptotic analysis of the PSS expressions shows that the previously used zero-order release assumption is not quite correct, even at later times, and this affects parameter estimation procedures. A comparison between the model of Bechard and McMullen and earlier obtained experimental data [J. Pharm. Sci. 72 (1983) 17] reveals some qualitative discrepancies that are yet to be explained. PMID- 11018549 TI - Galactosylated chitosan-graft-dextran as hepatocyte-targeting DNA carrier. AB - Lactobionic acid bearing galactose group was coupled with chitosan for liver specificity, and dextran was grafted to galactosylated chitosan (GC) for stability in water. Compared to the GC/DNA complex, the stability of the galactosylated chitosan-graft-dextran (GCD)/DNA complex could be enhanced. The particle size of the GCD/DNA complexes decreased as the charge ratio of GCD to DNA increased. Conformational change of DNA did not occur after complex formation with GCD compared with the conformation of DNA itself. The GCD/DNA complexes were only transfected into Chang liver cells and that of Hep G2 having asialoglycoprotein receptors (ASGR), indicative of specific interaction of ASGRs on cells and galactose ligands on chitosan. PMID- 11018551 TI - Gene expression control by temperature with thermo-responsive polymeric gene carriers. AB - A thermo-responsive copolymer, poly(N-isopropylacrylamide (IPAAm)-co-2 (dimethylamino)ethyl methacrylate (DMAEMA)-co-butylmethacrylate (BMA)), was synthesized and its in vitro gene transfection efficiency at different incubation temperatures was evaluated. A copolymer containing 8 mol% DMAEMA and 11 mol% BMA (P(IP-8DA-11BM)) had a lower critical solution temperature (LCST) at 21 degrees C, therefore the copolymer was insoluble above 21 degrees C and soluble below 21 degrees C. The LCST of P(IP-8DA-11BM) solution was not affected by the presence of salmon DNA. This copolymer was complexed with plasmid DNA, and the stability of the complex was analyzed by gel electrophoresis. DNA was completely retained in the complex, which was observed in the gel loading slot at 37 degrees C. At 20 degrees C, DNA was found to be partially dissociated from the complex by the appearance of the same band as DNA in the control experiment. These results clearly show that complex formation/dissociation was modulated by temperature alteration. The transfection efficiency of polymer-plasmid complexes was evaluated in COS-1 cells using pCMV-lacZ plasmid, encoding for beta-galactosidase as a reporter gene. The transfection efficiency of PDMAEMA homopolymer incubated at 37 degrees C for 48 h was greater than that incubated at 20 degrees C for 3 h and 37 degrees C for 45 h. In contrast, the transfection efficiency of P(IP-8DA 11BM) incubated at 20 degrees C for 3 h and 37 degrees C for 45 h was much higher than that incubated at 37 degrees C for 48 h. Such an increased transfection efficiency on lowering the temperature is considered to be due to appropriate formation/dissociation control of P(IP-8DA-11BM)-DNA complexes. PMID- 11018552 TI - Characteristics and biodistribution of cationic liposomes and their DNA complexes. AB - We have developed some novel liposome formulations for gene transfection. The formulations consisting of O,O'-ditetradecanoyl-N-(alpha-trimethyl ammonio acetyl) diethanolamine chloride (DC-6-14) as a cationic lipid, phospholipid and cholesterol showed effective gene transfection activity in cultured cells with serum and in vivo, i.e., intraperitoneal injection in mice. In this report, the physicochemical characteristics and biodistribution of the liposomes containing DC-6-14 (DC-6-14 liposomes) as a drug (gene) carrier for gene therapy were investigated in vitro and in vivo. DC-6-14 liposome-DNA complexes were usually thought to have positive surface charge. However, depending on the ratio of DNA to liposomes, zeta-potential of the complexes became negative. The diameter of the complexes also depended on the DNA-liposome ratio, and showed a maximum when their surface potential was neutral. When biodistribution of the complexes was determined after intravenous injection, positively charged complexes showed an immediate lung accumulation. On the other hand, negatively charged complexes did not show lung accumulation. These results have suggested that biodistribution of the DNA-liposome complexes, prepared with DC-6-14 liposomes, depends on their surface charge. Therefore, some surface modification of DC-6-14 liposomes may improve the biodistribution and hence the targetability of their DNA complexes. PMID- 11018553 TI - Controlled release of nerve growth factor from a heparin-containing fibrin-based cell ingrowth matrix. AB - The goal of this work was to develop a growth factor delivery system for use in nerve regeneration that would provide localized release of beta-nerve growth factor (beta-NGF) and other members of the neurotrophin family in a controlled manner. Although beta-NGF does not bind heparin with high affinity, we postulated that a basic domain found at the surface of native beta-NGF could interact with heparin and slow its diffusion from a heparin-containing delivery system. To test this hypothesis, we used a heparin-containing fibrin-based cell ingrowth matrix consisting of three components, namely an immobilized heparin-binding peptide, heparin and a neurotrophin with low heparin-binding affinity. The heparin-binding peptide contained a factor XIIIa substrate and was covalently cross-linked to fibrin matrices during polymerization. This cross-linked heparin-binding peptide served to immobilize heparin within the matrix, and this immobilized heparin interacted with the neurotrophin and slowed the passive release of the growth factor from the matrix. The ability of heparin-containing fibrin matrices, with a high excess of heparin-binding sites, to slow the diffusion-based release of beta NGF from fibrin matrices was measured in the absence of cells. Conditions that provided for slow diffusion-based release of beta-NGF, brain-derived neurotrophic factor, and neurotrophin-3 were tested in an assay of neurite extension from dorsal root ganglia to determine the ability of the delivery system to release active growth factor. The results demonstrated that neurotrophins, interacting with fibrin matrices containing a large molar excess of heparin relative to growth factor, enhanced neurite extension by up to 100% relative to unmodified fibrin. In the absence of the delivery system, free neurotrophins within the fibrin matrix did not enhance neurite extension. The results suggest that these matrices could serve as therapeutic materials to enhance peripheral nerve regeneration through nerve guide tubes and may have more general usefulness in tissue engineering for the delivery of non-heparin-binding growth factors. PMID- 11018554 TI - Fibrin encapsulated liposomes as protein delivery system. Studies on the in vitro release behavior. AB - The efficacy of biologically active proteins in medical therapy depends on the development of suitable drug delivery systems. These delivery systems need to overcome the severe problems connected with the use of proteins such as their usually short half lives in body fluids and their susceptibility to proteolysis and denaturation. Our delivery system combines two widespread devices by encapsulating liposomes containing the model protein horseradish peroxidase (HRP) inside the biopolymer fibrin. The liposomes enable the protein to remain in its preferred aqueous environment and protect it during the polymerization process. Further encapsulation of the liposomes inside fibrin was carried out in order to achieve a depot system with sustained protein release. In vitro experiments showed that the protein filled liposomes were absolutely stable within the fibrin network. In contrast to 'free' HRP, enzyme entrapped in liposomes was completely retained by the fibrin network and wasn't released from the device unless the fibrin was degraded by plasmin. PMID- 11018555 TI - Cross-linked hyaluronic acid hydrogel films: new biomaterials for drug delivery. AB - A new hyaluronic acid (HA)-based hydrogel film was prepared and evaluated for use in drug delivery. This biocompatible material crosslinks and gels in minutes, and the dried film swells and rehydrates to a flexible hydrogel in seconds. HA was first converted to the adipic dihydrazide derivative and then crosslinked with the macromolecular homobifunctional reagent poly(ethylene glycol) propiondialdehyde to give a polymer network. After gelation, a solvent casting method was used to obtain a HA hydrogel film. The dried film swelled sevenfold in volume in buffer, reaching equilibrium in less than 100 s. Scanning electron microscopy (SEM) of the hydrogel films showed a condensed and featureless structure before swelling, but a porous microstructure when hydrated. The thermal behavior of the hydrogel films was characterized by differential scanning calorimetry. The enzymatic degradation of the HA hydrogel films by hyaluronidase was studied using both SEM and a spectrophotometric assay. Drug release from the hydrogel film was evaluated in vitro using selected anti-bacterial and anti inflammatory drugs. This novel biomaterial can be employed for controlled release of therapeutic agents at wound sites. PMID- 11018556 TI - Time- and concentration-dependent apoptosis and necrosis induced by free and HPMA copolymer-bound doxorubicin in human ovarian carcinoma cells. AB - A2780 sensitive and A2780/AD doxorubicin (DOX) resistant human ovarian carcinoma cells were exposed to different concentrations (0.25, 0.5, 1, 5 and 10xIC(50)) of free and HPMA copolymer-bound DOX for 12, 24, 36, 48, 60 and 72 h. Apoptosis and necrosis were evaluated using the FITC-conjugated annexin V and propidium iodide staining. The data obtained showed that the induction of apoptosis and necrosis by both free DOX and HPMA copolymer-bound DOX were time- and concentration dependent. The data also showed significant differences between the drugs. It was found that: (i) under the action of HPMA copolymer-bound doxorubicin the alterations in the plasma membrane permeability preceded disturbances in cellular metabolism; (ii) HPMA copolymer-bound doxorubicin kills the cells mainly by necrosis; (iii) HPMA copolymer-bound doxorubicin is a more effective anticancer drug than free doxorubicin. PMID- 11018557 TI - Improvement of the encapsulation efficiency of oligonucleotide-containing biodegradable microspheres. AB - The objective of this study was to encapsulate an oligonucleotide drug within poly(lactide) microparticles with high encapsulation efficiencies at high theoretical drug loadings by the solvent evaporation method. With the conventional W/O/W method, the encapsulation efficiency decreased with increasing internal water content, increasing stirring time prior to filtration of the microparticles and increasing drug loading. The encapsulation was improved by replacing methylene chloride with ethyl acetate, by using micronized drug powder instead of an internal aqueous phase or by adding electrolytes or nonelectrolytes to the external phase. With ethyl acetate, a pre-emulsification step into a smaller volume of external aqueous phase was necessary in order to avoid premature polymer precipitation and to obtain microparticles. The addition of salts (NaCl or MgCl(2)) or sorbitol to the external aqueous phase significantly improved the encapsulation efficiency, even at high theoretical drug loadings. The microparticles had a denser structure with a smooth, pore-free surface. PMID- 11018558 TI - Microemulsions for topical delivery of 8-methoxsalen. AB - 8-Methoxsalen (8-MOP) and related furocumarins have been extensively used for the treatment of hyperproliferative skin diseases in association with long-wavelength UVA light. In order to develop alternative formulations for the topical administration of 8-MOP, microemulsions were evaluated as delivery vehicles. Six microemulsion formulations were prepared using water, isopropyl myristate (IPM) and Tween((R)) 80: Span((R)) 80: 1,2-Octanediol (3:1:1.2 w/w). The microemulsions were characterized using conductimetric and dynamic light scattering analyses. The ability of the systems to deliver 8-MOP into and through the skin was evaluated in vitro using newborn pig-skin. The in vitro permeation data showed that the novel microemulsions increased the 8-MOP total penetration through the skin by order of 1.9-4.5, as compared with IPM. In general, the accumulation of 8 MOP into the skin was increased by a factor of 1.5-4.5 by the microemulsion systems with respect to their total amount of drug delivered across the skin. These results suggest that the studied microemulsion systems may be appropriate vehicles for the topical delivery of 8-MOP. PMID- 11018560 TI - Fraud in medical research: an international survey of biostatisticians. ISCB Subcommittee on Fraud. AB - The characteristics of scientific fraud and its impact on medical research are in general not well known. However, the interest in the phenomenon has increased steadily during the last decade. Biostatisticians routinely work closely with physicians and scientists in many branches of medical research and have therefore unique insight into data. In addition, they have methodological competence to detect fraud and could be expected to have a professional interest in valid results. Biostatisticians therefore are likely to provide reliable information on the characteristics of fraud in medical research. The objective of this survey of biostatisticians, who were members of the International Society for Clinical Biostatistics, was to assess the characteristics of fraud in medical research. The survey was performed between April and July 1998. The participation rate was only 37%. We report the results because a majority (51%) of the participants knew about fraudulent projects, and many did not know whether the organization they work for has a formal system for handling suspected fraud or not. Different forms of fraud (e.g., fabrication and falsification of data, deceptive reporting of results, suppression of data, and deceptive design or analysis) had been observed in fairly similar numbers. We conclude that fraud is not a negligible phenomenon in medical research, and that increased awareness of the forms in which it is expressed seems appropriate. Further research, however, is needed to assess the prevalence of different types of fraud, as well as its impact on the validity of results published in the medical literature. PMID- 11018561 TI - A flexible stochastic curtailing procedure for the log-rank test. AB - For safety and ethical reasons, a data monitoring committee of a clinical trial may wish to assess the futility of continuing a trial if the currently available data at an interim look show no beneficial effect due to treatment, especially when accompanied by mounting evidence of treatment emergent adverse effects. Stochastic curtailing whereby conditional power is evaluated given currently observed data is one way of evaluating futility. In clinical trials that look at "time-to-event" as the primary outcome, difference between treatment groups with respect to the primary outcome is commonly evaluated using the log-rank test. Although the unconditional power function for the log-rank test has been described previously, its conditional power has not been widely investigated. We describe a method for evaluating conditional power when the log-rank test is used to assess the difference between the survival distributions of two treatment groups with respect to some failure-time outcome. The method is useful under a wide range of assumptions regarding the underlying survival distribution, patient entry distribution, losses to follow-up, and (if applicable) noncompliance, drop ins, lag in treatment effect, and stratification. This level of applicability is attained by generalizing a flexible Markov chain approach to unconditional power computation, described previously, to compute conditional power. PMID- 11018562 TI - Reengineering a database for clinical trials management: lessons for system architects. AB - This paper describes the process of enhancing Trial/DB, a database system for clinical studies management. The system's enhancements have been driven by the need to maximize the effectiveness of developer personnel in supporting numerous and diverse users, of study designers in setting up new studies, and of administrators in managing ongoing studies. Trial/DB was originally designed to work over a local area network within a single institution, and basic architectural changes were necessary to make it work over the Internet efficiently as well as securely. Further, as its use spread to diverse communities of users, changes were made to let the processes of study design and project management adapt to the working styles of the principal investigators and administrators for each study. The lessons learned in the process should prove instructive for system architects as well as managers of electronic patient record systems. PMID- 11018563 TI - Gender representation in trials. AB - The perception is that women have been understudied relative to men. It has been sufficient to cause Congress to enact legislation to require that a clinical trial must be "designed and carried out in a manner sufficient to provide for a valid analysis of whether the variables being studied in the trial affect women ellipsis differently than other subjects in the trial." We looked for evidence as to whether the perception has a basis in fact by looking at measures of gender based research effort. Clinical trials, published between 1966 and 1998 in U.S. journals and indexed in MEDLINE, were classified by gender. Reports of trials appearing in five widely circulated medical journals (Annals of Internal Medicine, British Medical Journal, Journal of the American Medical Association, Lancet, and New England Journal of Medicine) in 1985, 1990, and 1995 were retrieved and read to obtain counts of the numbers of males and females represented in trials published in those journals. For reports of trials published in U.S. journals, the percent involving males and females, males only, females only, and those where gender was not specified were 55.2%, 12.2%, 11.2%, and 21.4%, respectively. Counts of males and females represented in the reports of trials appearing in the five aforementioned journals were 355,624 and 550,743, respectively. We did not find evidence of systematic effort bias against females. PMID- 11018564 TI - Identifying clinical trials in the medical literature with electronic databases: MEDLINE alone is not enough. AB - The objective of this study was to compare the performance of MEDLINE and EMBASE for the identification of articles regarding controlled clinical trials (CCTs) published in English and related to selected topics: rheumatoid arthritis (RA), osteoporosis (OP), and low back pain (LBP). MEDLINE and EMBASE were searched for literature published in 1988 and 1994. The initial selection of papers was then reviewed to confirm that the articles were about CCTs and to assess the quality of the studies. Selected journals were also hand searched to identify CCTs not retrieved by either database. Overall, 4111 different references were retrieved (2253 for RA, 978 for OP, and 880 for LBP); 3418 (83%) of the papers were in English. EMBASE retrieved 78% more references than MEDLINE (2895 versus 1625). Overall, 1217 (30%) of the papers were retrieved by both databases. Two hundred forty-three papers were about CCTs. Two-thirds of these were retrieved by both databases, and one-third by only one. An additional 16 CCTs not retrieved by either database were identified through hand searching. Taking these into account, EMBASE retrieved 16% more CCTs than MEDLINE (220 versus 188); the EMBASE search identified 85% of the CCTs compared to 73% by MEDLINE. No significant differences were observed in the mean quality scores and sample size of the CCTs missed by MEDLINE compared to those missed by EMBASE. Our findings suggest that the use of MEDLINE alone to identify CCTs is inadequate. The use of two or more databases and hand searching of selected journals are needed to perform a comprehensive search. PMID- 11018565 TI - Indirect comparison in evaluating relative efficacy illustrated by antimicrobial prophylaxis in colorectal surgery. AB - This paper aims to explore the potential usefulness and limitations of indirect comparisons in evaluating the relative efficacy of interventions. From a systematic review of antimicrobial prophylaxis in colorectal surgery, we identified 11 sets of randomized trials that can be used to compare antibiotics both directly and indirectly. The discrepancy between the direct and the indirect comparison is defined as the absolute value of difference in log odds ratio. The adjusted indirect comparison has the advantages that the prognostic factors of participants in different trials can be partially taken into account and more uncertainty be incorporated into its result by providing a wider confidence interval. However, considerable discrepancies exist between the direct and the adjusted indirect comparisons. When there is no direct comparison, the adjusted indirect method may be used to obtain some evidence about the relative efficacy of competing interventions, although such indirect results should be interpreted with great caution. Further empirical and methodologic research is needed to explore the validity and generalizability of the adjusted indirect comparison for evaluating different interventions. PMID- 11018566 TI - Fraud is bad, studying fraud is hard. PMID- 11018567 TI - Design and statistical issues of the hemodialysis (HEMO) study. AB - The Hemodialysis Study is a multicenter clinical trial of hemodialysis prescriptions for patients with end stage renal disease. Participants from over 65 dialysis facilities associated with 15 clinical centers in the United States are randomized in a 2 x 2 factorial design to dialysis prescriptions targeted to a standard dose or a high dose, and to either low or high flux membranes. The primary outcome variable is mortality; major secondary outcomes are defined based on hospitalizations due to cardiovascular or infectious complications, and on the decline of serum albumin. The Outcome Committee, consisting of study investigators, uses a blinded review system to classify causes of death and hospitalizations related to the major secondary outcomes. The dialysis dose intervention is directed by the Data Coordinating Center using urea kinetic modeling programs that analyze results from dialysis treatments to monitor adherence to the study targets, adjust suggested dialysis prescriptions, and assist in trouble-shooting problems with the delivery of dialysis. The study design has adequate power to detect reductions in mortality rate equal to 25% of the projected baseline mortality rate for both of the interventions. PMID- 11018569 TI - Adherence to behavioral and pharmacological interventions in clinical research on older adults. PMID- 11018570 TI - Studying adherence to therapeutic regimens: overview, theories, recommendations. AB - Prediction of adherence and planning of behavior change can be systematically accomplished by using or modifying well-investigated theories. An overview of these models and recommendations for their use is offered. Control Clin Trials 2000;21:156S-163S PMID- 11018571 TI - An examination of theory and behavior change in randomized clinical trials. AB - This paper underscores the important role played by theory-based mediating variables in randomized clinical trials. Indeed, it is essential that we know what mediating variables are relevant for particular outcomes in randomized clinical trials and that we design interventions to optimize change in the mediators of interest. Yet, knowledge of the pivotal mediating variables in behavior change does not imply that we know how to intervene effectively. This gap may be due to the fact that existing research typically has been designed to garner support for theory, as opposed to testing support for behavior change strategies that are based on theory. In addition, we argue that there are important mediating variables in behavior change that have not been given systematic attention in theory development. For example, behavior change must be viewed as a collaborative process, and participants' perceptions concerning the feasibility of change is important to motivational processes in randomized clinical trials. Control Clin Trials 2000;21:164S-170S PMID- 11018572 TI - Issues of aging and adherence to health interventions. AB - This review article describes several processes and considerations that are important to adherence in aging research and potential strategies that could be used to facilitate adherence among older adults. In many large aging trials participants are purposefully selected to reduce the risk of suboptimal adherence and retention. This selection often involves screening out those with barriers such as transportation needs, sensory deficits, functional dependence, major diseases limiting life expectancy, or apparent psychological distress. However, trends toward extending interventions to the general population of older adults require specific knowledge about the circumstances and processes that support adherence among older adults or the conditions that make them vulnerable to adherence problems. Addressing the diversity of needs, expectations, and capabilities of older adults that promote adherence is a key consideration in aging research. Control Clin Trials 2000;21:171S-183S PMID- 11018573 TI - Adherence in social context. AB - Much has been written about adherence to interventions in older adults. What has not been discussed as extensively is how adherence is influenced and affected by the multiple interacting layers of the social context. Guided by an ecological or multilevel system model, this paper explores how social context may impact adherence. We conclude that when considering strategies to promote adherence to a particular regime, one must take into consideration the interplay between the social context and the adherence-related behavior. Control Clin Trials 2000;21:184S-187S PMID- 11018574 TI - Measuring adherence to behavioral and medical interventions. AB - Measuring adherence to medical and behavioral interventions is important to clinicians and researchers since inadequate adherence can reduce the effectiveness of an intervention. Unfortunately, there is no gold standard for measuring adherence across health behaviors. Adherence needs to be defined situationally with parameters of acceptable adherence carefully delineated and appropriate to the health behavior being studied. Additionally, measurement methods must be valid, reliable, and sensitive to change; this paper reviews these criteria. Methods used to measure adherence to dietary interventions include 24-hour recalls, food diaries, and food frequency questionnaires. Direct and indirect calorimetry, doubly labeled water, and a variety of self-report methods can be used to measure adherence in physical activity interventions. Adherence to pharmacological interventions is assessed using self-report methods, biochemical measures, medication counts, and the automated pharmacy database review strategy. The strengths and weaknesses of these methods for measuring adherence to dietary, physical activity, and pharmacological interventions are reviewed. Control Clin Trials 2000;21:188S-194S PMID- 11018575 TI - Who will adhere? Key issues in the study and prediction of adherence in randomized controlled trials. AB - Adherence determinants and key adherence research issues are discussed for three types of randomized controlled trials: pharmacological, dietary, and physical activity. This article highlights theoretical and methodological limitations that have hampered the ability to identify patients at risk for poor treatment compliance. Control Clin Trials 2000;21:195S-199S PMID- 11018576 TI - Predictors of adherence to behavior change interventions in the elderly. AB - Considering factors at the individual, interpersonal, and environmental level may enhance adherence to interventions in the elderly. A collaborative practitioner participant relationship is also essential. Control Clin Trials 2000;21:200S-205S PMID- 11018577 TI - Dietary adherence: characteristics and interventions. AB - This paper reviews issues regarding dietary adherence. Issues and barriers unique to dietary adherence, in contrast to adherence to physical activity or medication regimens, are discussed. These include decision making, social and cultural contexts, perceptions and preferences, and environmental barriers. We review factors known to increase adherence in dietary interventions, including education, motivation, behavioral skills, new and modified foods, and supportive interactions. We conclude with directions for future study, such as improved measurement of diet-related behavior and longitudinal, culturally sensitive interventions. Control Clin Trials 2000;21:206S-211S PMID- 11018578 TI - Exercise adherence among older adults: challenges and strategies. AB - This paper explores physical activity adherence among older adults in clinical research. We address the unique challenges associated with promoting physical activity, including the amount of time and effort, perceived benefits, and recommendations from professionals. We explore useful strategies to enhance adherence in the realm of physical activity interventions, including the development of theory-based physical activity interventions that acknowledge participant needs and expectations, promote self-regulatory skills, view exercise as an ongoing process subject to relapse, and utilize phone contacts for home based activity programs. We conclude with directions for future clinical trials to enhance exercise adherence that include the need for a precise definition and operationalization of adherence, linkages between physical activity interventions and theory, and the study of exercise in both structured and home-based settings. Control Clin Trials 2000;21:212S-217S PMID- 11018579 TI - Adherence to pharmacological interventions. Current trends and future directions. The Pharmacological Intervention Working Group. AB - The complexity of adherence to pharmacological interventions is frequently underestimated. Consequently, little research has been done to understand, improve, or evaluate the effects of adherence to drug protocols. While efforts to better measure and increase adherence require dedicated resources, improved adherence in existing clinical research can potentially cut costs in trial size and length. This paper outlines strategies to facilitate research on adherence to pharmacological interventions including: reporting of adherence data, changing how we conceptualize adherence to pharmaceuticals, understanding issues unique to older adults' pill-taking behavior, and creating standardized methodologies to measure adherence. Further, we describe some promising research areas that may lead to effective interventions: adherence "typologies" and modality matching. Control Clin Trials 2000;21:218S-225S PMID- 11018580 TI - Enhancing adherence in randomized controlled clinical trials. AB - This paper identifies individual and systematic primary, secondary, and tertiary prevention approaches that can be used to enhance adherence in randomized controlled trials. Specifically, strategies such as these are discussed: careful screening during enrollment, use of a run-in period, fully informing participants of the study burden, explaining randomization, determining the participant's prior history with treatment interventions, establishing and using a tracking system, clearly specifying adherence measures, and staff training in communication and negotiation. These strategies have been gathered over time from our experiences working in multiple randomized controlled trials. Research on factors that promote or detract from adherence should be a standard part of randomized controlled trials. Control Clin Trials 2000;21:226S-232S PMID- 11018581 TI - Informed adherence: the need for shared medical decision making. AB - Screening tests for colon cancer, breast cancer, and elevated serum cholesterol are widely advocated and included in some practice guidelines. Failure to complete these tests is regarded as patient noncompliance. The purpose of this paper is to review noncompliance with screening tests from the perspective of a traditional biomedical model and an outcomes model. According to the traditional biomedical model, compliance with screening tests is necessary to identify disease at an early stage and to initiate treatment. In contrast, the outcomes model argues that patients and providers should engage in activities that maximize benefit from the patient's perspective. Screening may lead to significant increases in case identification and in treatment rates. However, screening might also identify "pseudodisease," defined as disease that is not clinically important. For some diagnostic tests, cases of pseudodisease far exceed cases of clinically meaningful disease. The outcomes model suggests shared medical decision making in which uncertainties surrounding tests are disclosed to patients. Using information about the risks and expected health outcomes of screening and treatment, patients can participate in decisions about their own health care. Control Clin Trials 2000;21:233S-240S PMID- 11018582 TI - Ethics in adherence promotion and monitoring. AB - In evaluating and intervening to increase adherence to medical treatments, clinicians and researchers must address ethical issues pertaining to best interest, autonomy, and privacy. "Best interest" refers to the notion that health care practitioners act in a manner that produces benefits or good outcomes for the patients in their care. "Autonomy" refers to the patient's right to determine whether or not they will accept medical treatment or participate in a clinical study. "Nonmaleficence" refers to the clinician's or researcher's responsibility to "do no harm." "Privacy" refers to the notion that researchers and clinicians promise not to divulge personal information about the patients in their care. Adherence monitoring and promotion pose ethical challenges to researchers and clinicians, which are the topic of this paper. Control Clin Trials 2000;21:241S 247S PMID- 11018583 TI - Going APE over ref-1. AB - The DNA base excision repair (BER) pathway is responsible for the repair of cellular alkylation and oxidative DNA damage. A crucial and the second step in the BER pathway involves the cleavage of baseless sites in DNA by an AP endonuclease. The major AP endonuclease in mammalian cells is Ape1/ref-1. Ape1/ref-1 is a multifunctional protein that is not only responsible for repair of AP sites, but also functions as a reduction-oxidation (redox) factor maintaining transcription factors in an active reduced state. Ape1/ref-1 has been shown to stimulate the DNA binding activity of numerous transcription factors that are involved in cancer promotion and progression such as Fos, Jun, NF(B, PAX, HIF-1(, HLF and p53. Ape1/ref-1 has also been implicated in the activation of bioreductive drugs which require reduction in order to be active and has been shown to interact with a subunit of the Ku antigen to act as a negative regulator of the parathyroid hormone promoter, as well as part of the HREBP transcription factor complex. Ape1/ref-1 levels have been found to be elevated in a number of cancers such as ovarian, cervical, prostate, rhabdomyosarcomas and germ cell tumors and correlated with the radiosensitivity of cervical cancers. In this review, we have attempted to try and assimilated as much data concerning Ape1/ref 1 and incorporate the rapidly growing information on Ape1/ref-1 in a wide variety of functions and systems. PMID- 11018584 TI - Characterization of the hOGG1 promoter and its expression during the cell cycle. AB - The human OGG1 gene codes for a 38kD protein with an antimutator activity related to its capacity to excise the mutagenic base 8-OH-Guanine from DNA. Mutant forms of this gene have been found in lung and kidney tumors. The determination of the start of transcription allowed the definition of the promoter sequences for the gene. By transient transfection and a luciferase reporter assay a 135 base pair region immediately upstream of the transcription start is shown to have full promoter activity. Two CpG islands and an Alu repeat were identified within the promoter and the 5' sequences of the transcribed region. The lack of TATA or CAAT boxes suggests that OGG1 is a housekeeping gene. Consistently, its expression, measured as the transcription from the promoter or as the enzymatic activity in cultured fibroblast cell lines, does not vary during the cell cycle. PMID- 11018585 TI - Transcriptional effects on double-strand break-induced gene conversion tracts. AB - Transcription stimulates spontaneous homologous recombination, but prior studies have not investigated the effects of transcription on double-strand break (DSB) induced recombination in yeast. We examined products of five ura3 direct repeat substrates in yeast using alleles that were transcribed at low or high levels. In each strain, recombination was stimulated by DSBs created in vivo at an HO site in one copy of ura3. Increasing transcription levels in donor or recipient alleles did not further stimulate DSB-induced recombination, nor did it alter the relative frequencies of conversion and deletion (pop-out) events. This result is consistent with the idea that transcription enhances spontaneous recombination by increasing initiation. Gene conversion tracts were measured using silent restriction fragment length polymorphisms (RFLPs) at approximately 100bp intervals. Transcription did not alter average tract lengths, but increased transcription in donor alleles increased both the frequency of promoter-proximal (5') unidirectional tracts and conversion of 5' markers. Increased transcription in recipient alleles increased the frequency of bidirectional tracts. We demonstrate that these effects are due to transcription per se, and not just transcription factor binding. These results suggest that transcription influences aspects of gene conversion after initiation, such as strand invasion and/or mismatch repair (MMR). PMID- 11018586 TI - Alteration of ultraviolet-induced mutagenesis in yeast through molecular modulation of the REV3 and REV7 gene expression. AB - DNA damage can lead to mutations during replication. The damage-induced mutagenesis pathway is an important mechanism that fixes DNA lesions into mutations. DNA polymerase zeta (Pol zeta), formed by Rev3 and Rev7 protein complex, and Rev1 are components of the damage-induced mutagenesis pathway. Since mutagenesis is an important factor during the initiation and progression of human cancer, we postulate that this mutagenesis pathway may provide an inhibiting target for cancer prevention and therapy. In this study, we tested if UV-induced mutagenesis can be altered by molecular modulation of Rev3 enzyme levels using the yeast Saccharomyces cerevisiae as a eukaryotic model system. Reducing the REV3 expression in yeast cells through molecular techniques was employed to mimic Pol zeta inhibition. Lower levels of Pol zeta significantly decreased UV-induced mutation frequency, thus achieving inhibition of mutagenesis. In contrast, elevating the Pol zeta level by enhanced expression of both REV3 and REV7 genes led to a approximately 3-fold increase in UV-induced mutagenesis as determined by the arg4-17 mutation reversion assays. In vivo, UV lesion bypass by Pol zeta requires the Rev1 protein. Even overexpression of Pol zeta could not alleviate the defective UV mutagenesis in the rev1 mutant cells. These observations provide evidence that the mutagenesis pathway could be used as a target for inhibiting damage-induced mutations. PMID- 11018587 TI - Oxidative DNA base damage induced by singlet oxygen and photosensitization: recognition by repair endonucleases and mutagenicity. AB - We have analyzed the recognition by various repair endonucleases of DNA base modifications induced by three oxidants, viz. [4-(tert butyldioxycarbonyl)benzyl]triethylammonium chloride (BCBT), a photochemical source of tert-butoxyl radicals, disodium salt of 1,4-etheno-2,3-benzodioxin-1,4 dipropanoic acid (NDPO(2)), a chemical source of singlet oxygen, and riboflavin, a type-I photosensitizer. The base modifications induced by BCBT, which were previously shown to be mostly 7,8-dihydro-8-oxoguanine (8-oxoGua) residues, were recognized by Fpg and Ogg1 proteins, but not by endonuclease IIII, Ntg1 and Ntg2 proteins. In the case of singlet oxygen induced damage, 8-oxoGua accounted for only 35% of the base modifications recognized by Fpg protein. The remaining Fpg sensitive modifications were not recognized by Ogg1 protein and relatively poor by endonuclease III, but they were relatively good substrates of Ntg1 and Ntg2. In the case of the damage induced by photoexcited riboflavin, the fraction of Fpg sensitive base modifications identified as 8-oxoGua was only 23%. In contrast to the damage induced by singlet oxygen, the remaining lesions were not only recognized by Ntg1 and Ntg2 proteins and (relatively poor) by endonuclease III, but also by Ogg1 protein. The analysis of the mutations observed after transfection of modified plasmid pSV2gpt into Escherichia coli revealed that all agents induced near exclusively GC-->TA and GC-->CG transversions, the numbers of which were correlated with the numbers of 8-oxoGua residues and Ntg-sensitive modifications, respectively. In conclusion, both singlet oxygen and the type-I photosensitizer riboflavin induce predominantly oxidative guanine modifications other than 8-oxoGua, which most probably give rise to GC-->CG transversions and in which eukaryotic cells are substrates of Ntg1 and Ntg2 proteins. PMID- 11018589 TI - Reflecting on the modern industrial life-science environment. AB - 'The barriers between 'silos' in pharma R&D workflows are slowly eroding and this is driving a wider exchange of information across the various R&D disciplines' PMID- 11018588 TI - Uracil incorporation into a gene targeting construct reduces the frequency of homologous and nonhomologous recombinants in human cells. AB - Gene targeting allows the introduction of specific modifications into the eukaryotic genome by homologous recombination, but its efficiency is low in many mammalian systems. We are exploring different ways to increase the efficiency of gene targeting and we report here the effect of uracil incorporation in the targeting construct. Plasmids containing uracil substituting for a fraction of thymine residues are hyperrecombinogenic in some bacterial systems. To test whether a similar stimulation of recombination occurs in mammalian cells, we have prepared a uracil-rich HPRT targeting construct and quantified its homologous and nonhomologous recombination frequencies compared to the same plasmid lacking uracil. The uracil-rich plasmid led to reductions in both homologous and nonhomologous recombination in human cells. PMID- 11018590 TI - Poetry and verse: an ideal medium for scientific communication? AB - 'Many professional scientists over the years have expressed their thoughts and ideas in poetry.' PMID- 11018592 TI - Startling new impetus for schizophrenia research. PMID- 11018591 TI - Relaxin: a potential new treatment for vasoconstrictive disorders. PMID- 11018593 TI - Modulating AMPA receptors: key to mild cognitive impairment and memory? PMID- 11018595 TI - Predicting human safety: screening and computational approaches. AB - Current preclinical safety evaluation programs use a combination of computational methods, mechanistic in vitro screening and - primarily - in vivo experimentation to predict human toxicity. The rapid transition of pharmaceutical R&D into electronic R&D (e-R&D) makes it imperative that predictive safety testing also develops into an information-rich, knowledge-based process in the near future. Accordingly, enhanced databases and computational tools are expected to change the way the pharmaceutical industry assesses drug toxicity during discovery and early development. Expert use of prediction tools should lead to lower failure rates in drug development and decrease the cost and time involved in successful drug approval. PMID- 11018596 TI - Corrigendum. PMID- 11018597 TI - Endothelin-receptor antagonists: current and future perspectives. AB - Despite much effort over recent years to design and develop endothelin-receptor antagonists, these compounds are far from becoming new drug entities. This article will review preclinical data on select endothelin-receptor antagonists as well as clinical data on bosentan, the only molecule currently in Phase III clinical trials. Though efficacious, bosentan is less potent than the angiotensin converting enzyme (ACE) inhibitor, enalapril, in patients with hypertension. We will therefore discuss the possible reason(s) for this low potency, the consequences thereof, and a few therapeutic areas where endothelin-receptor antagonists could find better use. PMID- 11018598 TI - Development of novel nucleoside analogues for use against drug resistant strains of HIV-1. AB - Nucleoside analogue inhibitors of the reverse transcriptase (RT) enzyme of HIV-1 were the first class of compounds to be used in anti-HIV-1 therapy and are a cornerstone in highly active antiretroviral therapy. Despite the number of inhibitors of HIV-1 RT available for clinical use at the present time and the effectiveness of these compounds in combination regimens, long-term exposure of patients to these drugs often results in the development of viral resistance or long-term toxicity. For this reason, efforts to identify new agents with activity against drug-resistant strains of HIV-1 and with a toxicity profile that allows for individual patient tolerance of the drugs are still warranted. PMID- 11018599 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 11018600 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 11018601 TI - Bladder-sparing strategies for transitional cell carcinoma. PMID- 11018602 TI - A positive caver map response poorly predicts recovery of potency after radical prostatectomy. AB - OBJECTIVES: To determine whether preservation of the neurovascular bundles (NVBs), defined by a positive CaverMap response, correlates with the recovery of potency after radical prostatectomy. METHODS: We studied a group of 60 men who were potent preoperatively who underwent radical retropubic prostatectomy by one surgeon. The CaverMap was used after removal of the prostate to assess the integrity of the NVBs. Postoperative potency was assessed by a postal questionnaire and telephone interview, administered independently of the treating surgeon. Men were considered potent if they reported postoperative erections consistently sufficient for vaginal penetration with or without the use of sildenafil. RESULTS: The mean patient age was 59 years, and the median follow-up was 365 days. A positive CaverMap response was obtained in 73 (77%) of the 95 NVBs tested. The overall potency rate was 18%. No patients with a bilateral negative CaverMap response were potent, and 2 (22%) of 9 with a unilateral CaverMap response (negative versus unilateral response, P = 0.46) and 6 (27%) of 22 with bilateral CaverMap responses (negative versus bilateral response, P = 0.32) were potent. CONCLUSIONS: A positive CaverMap response, suggesting that a successful nerve-sparing prostatectomy had been performed, was obtained in 77% of the NVBs tested. Nevertheless, with a median follow-up of 12 months, most patients with a positive CaverMap response remained impotent. This suggests that other factors are critical to the recovery of sexual function after radical prostatectomy. PMID- 11018603 TI - Clinical significance of detrusor sphincter dyssynergia type in patients with post-traumatic spinal cord injury. AB - OBJECTIVES: To investigate the significance of categorizing detrusor sphincter dyssynergia (DSD) by type in patients with chronic spinal cord injury. METHODS: A retrospective review of the charts, video-urodynamic studies, and upper tract radiographic studies of 269 patients with post-traumatic, suprasacral spinal cord injuries was performed. The patients were categorized according to the DSD type (intermittent or continuous), level and completeness of injury, intravesical pressure at leak, upper tract complications, and interval since injury. RESULTS: Of the 269 patients, 20 (7.4%), 216 (80.3%), and 33 (12.3%) had no DSD, intermittent DSD, and continuous DSD, respectively. No significant association between the specific level of injury and the DSD type was found (P = 0.71). The presence of DSD was associated with complete injuries, elevated intravesical pressures, and upper tract complications (P <0.01); these associations were more prominent with continuous DSD than with intermittent DSD. The proportion of patients with no DSD, intermittent DSD, and continuous DSD was unchanged during the chronic follow-up period. CONCLUSIONS: The clinical significance of DSD type is not crucial, since patients with both intermittent and continuous DSD require urodynamic surveillance and expedient treatment to minimize urologic complications. However, the presence of continuous DSD is one of several factors that may require earlier urodynamic follow-up. PMID- 11018604 TI - Editorial comment. Detrusor-external sphincter dyssynergia. PMID- 11018605 TI - Reply by the authors PMID- 11018606 TI - Polymerase chain reaction in clinically suspected genitourinary tuberculosis: comparison with intravenous urography, bladder biopsy, and urine acid fast bacilli culture. AB - OBJECTIVES: To evaluate the role of urinary polymerase chain reaction (PCR) in the detection of Mycobacterium tuberculosis (MTb) in patients with a clinical suspicion of genitourinary tuberculosis (GUTB) and to compare its sensitivity with intravenous urography (IVU), bladder biopsy, and urine culture for acid fast bacilli (AFB). METHODS: The study was carried out between September 1997 and December 1998 in 42 patients with a clinical suspicion of GUTB. Their clinical features, organ involvement, and investigation results were studied. The diagnostic yield of urinary PCR for MTb and its sensitivity in comparison with routine urine AFB culture, bladder biopsy, and IVU were assessed. RESULTS: There were 25 male and 17 female patients, with a mean age of 31.04 years. Patients suspected of having GUTB most often presented with irritative voiding symptoms. Two patients had abnormal renal parameters. Of the 42 patients clinically suspected of having GUTB, radiologic abnormalities suggestive of GUTB were found in 37 (88.09%); MTb was isolated in the urine AFB culture in 13 (30.95%); bladder biopsy was positive in 11 (45.83%); and urinary PCR for MTb was positive in 34 cases (80.95%). Of 35 cases of proven GUTB, IVU was suggestive of the diagnosis in 32 (91.42%) and MTb was isolated in the urine AFB culture in 13 cases (37.14%). Bladder biopsy was positive in 11 (45. 83%) of 24 patients in whom biopsy was taken, and urinary PCR for MTb was positive in 33 (94.29%). CONCLUSIONS: A high index of suspicion is necessary for a diagnosis of GUTB. In clinically suspected cases, IVU may be suggestive of GUTB, but it is not specific. In the present study, IVU was suggestive in 88.09% of patients. MTb was isolated in the urine AFB culture in only 37.14% of patients, and bladder biopsy was positive in 45.83%. Urinary PCR for MTb was the most sensitive indicator and was positive in 94.29% of patients. It is evident from this series that PCR provides a much faster diagnosis of urinary MTb. It is a rapid, sensitive, and specific diagnostic method and avoids a delay in starting treatment. PMID- 11018607 TI - Intensive medical management of ureteral calculi. AB - OBJECTIVES: To compare two treatment regimens in patients with ureteral calculi. One regimen (control arm) used routine drugs, and the second regimen (treatment arm) used the same routine drugs plus uncommonly used drugs. METHODS: Between February and October 1998, 70 consecutive patients were evaluated for symptomatic ureteral calculi. Thirty-five patients were randomized to a control arm and received ketorolac, oxycodone, and acetaminophen combination tablets and prochlorperazine suppositories. Thirty-five patients were randomized to the treatment arm and received the same medications plus nifedipine XL, prednisone, and trimethoprim/sulfa combination tablets and plain acetaminophen. Stone passage rates, work days lost, emergency room visits, surgical interventions, and possible side effects of the drugs were recorded. RESULTS: The treatment arm (addition of nifedipine XL, prednisone, trimethoprim/sulfa, and plain acetaminophen) had higher (86% versus 56%) stone passage rates and fewer lost work days (mean 1.76 versus 4.9), emergency room visits (1 versus 4), and surgical interventions (2 versus 15). Both arms exhibited similar potential drug side effects. CONCLUSIONS: The addition of a calcium channel blocking agent, steroids, antibiotics, and more acetaminophen effected a higher stone passage rate and fewer lost work days, emergency room visits, and surgical interventions. PMID- 11018608 TI - Effectiveness of nifedipine and deflazacort in the management of distal ureter stones. AB - OBJECTIVES: To evaluate the effectiveness of medical therapy during watchful waiting in patients with distal ureter stones. METHODS: Ninety-six patients with radiopaque stones located in the distal tract of the ureter and with stone sizes of 1 cm or smaller were involved in the study. The patients were randomly divided into two groups. Group A (n = 48) received oral treatment with 30 mg of deflazacort daily (maximum 10 days) plus 30 mg of slow-release nifedipine daily (maximum 4 weeks). Group B (n = 48) underwent a wait-and-watch approach. Both groups of patients were allowed to use diclofenac on demand. Statistical analyses were carried out using Student's t test, the chi-square test, and Fisher's exact test. RESULTS: The average stone size was 5.8 +/- 1.8 mm for group A and 5. 5 +/- 1.4 mm for group B. No statistically significant difference was found in stone size. Stone expulsion was observed in 38 (79%) of 48 patients in group A and in 17 (35%) of 48 patients in group B. The average expulsion time was 7 days (range 2 to 10) for group A and 20 days (range 10 to 28) for group B. A statistically significant difference was observed in both the expulsion rate and the expulsion time (P <0.05). The mean amount of sodium diclofenac used was 15 mg per patient for group A and 105 mg per patient for group B (P <0.05). CONCLUSIONS: The medical treatment proved to be effective and safe, as demonstrated by the increased stone expulsion rate, decreased expulsion time, and reduced need for analgesic therapy. PMID- 11018609 TI - Editorial comment. The value of intensive medical management of distal ureteral calculi in an effort to facilitate spontaneous stone passage. PMID- 11018610 TI - Consecutive quantitative cytology in bladder cancer. AB - OBJECTIVES: Quantitative cytology (Quanticyt) provides an objective reproducible alternative for routine cytology. To increase the sensitivity of cytology, we studied the application of consecutive cytology in patients in follow-up for superficial bladder cancer. METHODS: Between 1991 and 1998, a set of five or more bladder wash samples was obtained from 614 patients. These patients were retrospectively studied for follow-up data. Each sample was scored according to the Quanticyt risk score. RESULTS: In 614 patients (508 men, 106 women), 5832 bladder wash samples were taken. The mean interval between the first and fifth sample per patient was 21.5 months (SD 13.8 months). The mean number of tumor recurrences per patient was 2.17 (SD 1.82). The risk score of the first sample was not predictive of recurrence. Invasive disease was found in 0%, 0%, and 0.8% of patients with one low, intermediate, and high-risk sample, respectively. After five samples, the corresponding rate was 0%, 0%, and 10%. A comparison of visual cytology and quantitative cytology revealed that the false-negative rate was significantly different (17% versus 3.8%). The positive predictive value was highest for visual cytology (17% versus 9.2%). CONCLUSIONS: Adding consecutive quantitative cytology to urine cytopathologic evaluation improves the detection rate of high-grade lesions. Combining quantitative cytology and visual cytology provided a more accurate prediction of tumor stage. PMID- 11018611 TI - Urinary tract erosions after synthetic pubovaginal slings: diagnosis and management strategy. AB - OBJECTIVES: To review our experience with the diagnosis and management of genitourinary tract erosions after pubovaginal sling placement. METHODS: Clinic and operative records from the urology and gynecology services at two university hospitals were reviewed, and 14 patients were identified who underwent surgical treatment for a urogenital tract erosion after pubovaginal sling placement. The presenting symptoms, physical findings, diagnostic procedures, surgical treatments, and outcomes were reviewed. RESULTS: Six vaginal erosions, six urethral and vaginal erosions, and two bladder erosions occurred. All were associated with synthetic sling or suture materials. Common symptoms included vaginal and urethral pain, irritative voiding symptoms, vaginal discharge and/or bleeding, and recurrent urinary tract infections. All vaginal and urethral erosions were detected by physical examination and cystoscopy. Symptoms resolved after removal of the eroded sling component. Of the 12 patients with vaginal or urethral erosions, 7 developed recurrent postoperative stress incontinence. CONCLUSIONS: Persistent painful or irritative symptoms after pubovaginal sling placement may be due to urogenital tract erosion, especially if synthetic materials were used. Appropriate evaluation and treatment will result in dramatic symptomatic improvement, although recurrent stress incontinence may occur. PMID- 11018612 TI - Editorial comment. Experience with synthetic slings. PMID- 11018613 TI - Cadaveric fascia lata sling: analysis of five recent adverse outcomes. AB - OBJECTIVES: To report a series of adverse outcomes of urethral sling procedures that used allograft fascia lata and to review the literature regarding the use of this material in genitourinary reconstruction. METHODS: Five neurologically normal patients presented to our center between August 1999 and October 1999 with complaints of recurrent incontinence or voiding dysfunction after undergoing urethral sling procedures at outside institutions that used fascia lata allografts. All patients underwent a thorough evaluation, including history and physical examination, voiding cystourethrography, and urodynamic studies. Sections of fascia were removed for histology in 2 patients at the time of operation. RESULTS: Three patients were diagnosed with de novo bladder outlet obstruction, 1 with recurrent urethral hypermobility, and 1 with intrinsic sphincteric deficiency. Histology of cadaveric fascia demonstrated collagen with almost complete absence of cellularity. There was no evidence of capillary or fibroblast ingrowth. All patients underwent reoperation and have had clinical improvement with short-term follow-up. CONCLUSIONS: Although distressing, the present series of adverse outcomes may simply reflect an overall increase in the number of sling procedures being performed nationally. Although decreased operative time and morbidity have been attributed to the use of fascia lata in urethral sling procedures, it is essential to ensure that long-term safety and efficacy will not be jeopardized before accepting it as a new standard of care. PMID- 11018614 TI - Comparison of artificial urinary sphincter and collagen for the treatment of postprostatectomy incontinence. AB - OBJECTIVES: To compare how urinary symptoms affect the quality of life in groups of men with postprostatectomy incontinence treated with collagen versus artificial urinary sphincter implantation. METHODS: Two cohorts of men, one which received urethral collagen injection and one artificial urinary sphincter (AUS) implantation, were surveyed with a validated quality-of-life questionnaire to assess how their urinary dysfunction impacted their daily activities. The mean impact score and bother score for the two groups were compared. In addition, the overall degree of continence between the groups was assessed. RESULTS: At a mean follow-up of 19 months, 8 (20%) of 41 patients treated with collagen injections were at least socially continent, requiring one pad daily or less. In comparison, 27 (75%) of 36 men treated with an AUS were at least socially continent (P <0.001). Both the impact score and the bother score from the quality-of-life questionnaire were significantly lower in the group treated with the AUS than in the group treated with collagen. CONCLUSIONS: Patients receiving an AUS achieved significantly higher continence rates. Also, the quality of life of men treated with an AUS was improved compared with that of the men treated with collagen injection. PMID- 11018615 TI - Long-term follow-up of laser treatment for lower urinary tract symptoms suggestive of bladder outlet obstruction. AB - OBJECTIVES: The long-term results of different laser technologies in the management of lower urinary tract symptoms (LUTS) suggestive of bladder outlet obstruction (BOO) are not well known. We studied the durability of the effect of laser prostatectomy and tried to identify the factors predictive of treatment outcome. METHODS: Between December 1992 and November 1996, 190 patients underwent laser prostatectomy because of LUTS suggestive of BOO. One hundred seven patients received visual laser ablation of the prostate (VLAP), 30 received contact laser vaporization (CLV), and 53 received interstitial laser coagulation (ILC). The baseline evaluation included the International Prostate Symptom Score (IPSS), uroflowmetry (maximum urinary flow rate), postvoid residual urine (PVR), prostate volume measurement, and urodynamic investigation. Patients were followed up until April 1999. Kaplan-Meier plots were constructed to calculate the risk of retreatment, and the log-rank test was used to evaluate the predictive value of clinical parameters for treatment failure. RESULTS: The median follow-up in the VLAP group was 53 months; the retreatment rate was 14% (95% confidence interval [CI] 6% to 22%). The corresponding numbers for the CLV and ILC groups were 47 months and 14% (95% CI 1% to 26%) and 34 months and 41% (95% CI 23% to 60%), respectively. A high PVR and a high grade of obstruction in the VLAP group, and a younger age in the ILC group, were associated with increased retreatment risk. CONCLUSIONS: VLAP and CLV have a durable effect, as demonstrated by their low retreatment rate. ILC is a less aggressive procedure, at the expense of a high retreatment rate. Patient selection for VLAP can be based on the grade of obstruction and PVR. PMID- 11018616 TI - Impact of baseline symptom severity on future risk of benign prostatic hyperplasia-related outcomes and long-term response to finasteride. The Pless Study Group. AB - OBJECTIVES: To evaluate the long-term effects of finasteride on symptoms, acute urinary retention (AUR), and the need for benign prostatic hyperplasia (BPH) related surgery in relationship to baseline symptom severity. METHODS: A total of 3040 men with BPH were treated for 4 years with finasteride or placebo. The changes from baseline in symptoms and the incidence of BPH-related surgery and AUR were determined in men with mild (less than 8), low-moderate (8 to 12), high moderate (13 to 19), and severe (greater than 19) baseline quasi-American Urological Association symptoms for all patients and for the subgroup with a baseline prostate-specific antigen (PSA) level of 1.4 ng/mL or greater. RESULTS: In patients who completed the 4-year study, the change in symptom score, stratified by baseline symptom severity, was +1.4 +/- 0.5 (mild), -0.8 +/- 0.3 (low-moderate), -3.6 +/- 0.3 (high-moderate), and -7.7 +/- 0.5 (severe) in finasteride-treated patients and, respectively, +3.4 +/- 0.5, +0.7 +/- 0.3, -1.4 +/- 0.3, and -5.3 +/- 0.6 in placebo-treated patients (between-group P <0.01). The between-group differences were greater in the subgroup of patients with a baseline PSA of 1.4 ng/mL or greater. The risk of BPH-related surgery increased among placebo patients with increasing baseline symptom severity to a greater extent than the risk of AUR. Finasteride reduced the risk of AUR or the need for BPH-related surgery in all subgroups (P <0.001), especially in men with a baseline PSA of 1.4 ng/mL or greater. CONCLUSIONS: Compared with placebo, finasteride had a beneficial effect on symptoms, AUR, and BPH-related surgery in all symptom categories. BPH-related surgery, but not AUR, occurred more commonly in placebo-treated men with more severe baseline symptoms. The greatest absolute benefit of finasteride on symptoms and the reduction in risk of AUR and surgery was in men with higher baseline symptom scores and a baseline PSA level of 1.4 ng/mL or greater. PMID- 11018617 TI - Detection of prostate cancer: a comparative study of the diagnostic efficacy of sextant transrectal versus sextant transperineal biopsy. AB - OBJECTIVES: The optimal biopsy strategy for the detection of prostate cancer still needs to be established, since a considerable proportion of clinically significant cancers remains undiagnosed on routine sextant transrectal biopsy. To assess the efficacy of transperineal biopsy to detect prostate cancer, we compared this approach to systematic sextant transrectal biopsy in a simulation experiment. METHODS: Ultrasound-guided sextant transverse (transrectal) biopsy and subsequent sextant longitudinal (transperineal) biopsy were performed on 40 radical prostatectomy specimens of patients with (transrectal) biopsy-detected prostate cancer. Conditions were simulative and may not be completely analogous to clinical settings. Ultrasound-determined prostate volume, biopsy tumor involvement, number of cores with cancer, and tumor volume were determined. Detailed mapping of radical prostatectomy specimens provided insight into the representativeness of the biopsy techniques. RESULTS: Of 40 cancers, 33 (82.5%) were redetected by the transperineal approach; 29 (72.5%) were detected by repeated transrectal biopsies. For both approaches, the tumor volume of the undiagnosed cancers was significantly smaller (P <0.01) and the prostate volume was significantly larger (P <0.01) than in the redetected ones. Between the two approaches, no difference was found for either of the variables determined in the redetected cancers. Prostate maps clarified that transperineal undiagnosed tumors were either small (0.2 cm(3) or less) or notably located at the prostatic base. CONCLUSIONS: The biopsy procedure in which the biopsy needles enter the prostate at the apex for a longitudinal direction may efficiently sample the prostatic peripheral zone. Since the experiment was artificial in design, caution should be observed in extrapolating these results to patient settings. PMID- 11018618 TI - Additional treatments and reimbursement rates associated with prostate cancer treatment for patients undergoing radical prostatectomy, interstitial brachytherapy, and external beam radiotherapy. AB - OBJECTIVES: To define patterns of treatment among contemporary patients undergoing radical prostatectomy, interstitial radiation, and external beam radiation for prostate cancer. METHODS: We analyzed 291 consecutive patients (Stage T1-T3NXM0) who underwent definitive local treatment for prostate cancer with radical prostatectomy, interstitial seed implantation, or external beam radiation. Patients were stratified into three risk groups based on clinical T stage, serum prostate-specific antigen level at diagnosis, and biopsy Gleason score. The frequency of additional treatments, including androgen deprivation and external beam radiation, given within 3 months of initial local therapy was assessed. Patterns of care were compared and adjusted for risk. RESULTS: Of the 291 patients, 107 (36. 8%) underwent radical prostatectomy, 94 (32.3%) underwent interstitial seed implantation, and 90 (30.9%) underwent external beam radiation. Use of combination therapy differed significantly according to the type of initial local treatment and risk category. No patient in the low-risk group received combination therapy. For patients in the intermediate and high-risk groups, the frequency of combination therapy was significantly lower in the radical prostatectomy group when compared with either the interstitial seed implantation (P <0.001 and P <0.02, respectively) or external beam radiation group (P <0.001 and P <0.001, respectively). CONCLUSIONS: There are significant differences in resource utilization for contemporary patients undergoing definitive local therapy for prostate cancer. These differences may have a significant effect on treatment cost and morbidity, and they will likely make short-term comparisons between different treatment modalities difficult because of the high use of androgen deprivation in men treated with radiation therapy. PMID- 11018619 TI - Effect of preoperative biofeedback/pelvic floor training on continence in men undergoing radical prostatectomy. AB - OBJECTIVES: To determine whether preoperative biofeedback training improves urinary continence overall or the rate of return of continence in men undergoing radical prostatectomy. METHODS: One hundred men scheduled to undergo radical prostatectomy were randomized to receive graded pelvic muscle exercise training with biofeedback 2 to 4 weeks before surgery or to a control group performing pelvic muscle exercises without biofeedback. The biofeedback group was instructed to continue exercises four times per day until surgery and to resume exercises when the urethral catheter was removed following surgery. The control group received written and brief verbal instructions in pelvic muscle exercises before surgery and again after catheter removal. Urinary continence was assessed by personal or phone interviews. RESULTS: Six months following surgery, the continence rates, as defined by the use of one pad or less per day, were 94% (44 of 47) and 96% (48 of 50) in the biofeedback and control groups, respectively (P = 0.596). Also, the rate of return as determined at time points 1, 2, 3, and 4 months after surgery was not significantly different between the two groups. CONCLUSIONS: Preoperative biofeedback training did not improve the outcome of pelvic muscle exercises on overall continence or the rate of return of urinary control in men undergoing radical prostatectomy. PMID- 11018620 TI - Sildenafil in the treatment of erectile dysfunction after radical prostatectomy. AB - OBJECTIVES: To evaluate the efficacy of sildenafil for the treatment of erectile dysfunction after radical prostatectomy and to determine whether age, preservation of the neurovascular bundles (NVBs), or the interval between surgery and the initiation of sildenafil therapy influences the response to sildenafil. METHODS: We began this study in April 1998, immediately after the Food and Drug Administration approved sildenafil. We surveyed 170 men who had undergone radical retropubic prostatectomy, had not recovered natural erections sufficient for intercourse, and subsequently received sildenafil between 3 and 24 months postoperatively. The data were collected through a confidential mail survey conducted by a clinical nurse. The men used a dose of 50 mg sildenafil and increased this to 100 mg if they did not obtain an adequate response. RESULTS: In the 120 men who began taking sildenafil at least 12 months after surgery, the overall response rate was 29%. Results varied markedly by patient age and number of NVBs preserved. In men younger than 55 years in whom both NVBs had been preserved, the response rate was 80%. In contrast, no patient older than 55 years in whom only one NVB had been preserved reported an adequate response. Regardless of age, no patient in whom both NVBs had been excised reported success with sildenafil. Of the 50 patients who began taking sildenafil less than 9 months after surgery and who had not recovered natural sexual function, none reported erections adequate for intercourse using sildenafil. CONCLUSIONS: Sildenafil is an effective treatment for men with erectile dysfunction after radical retropubic prostatectomy, particularly in younger men in whom both NVBs have been preserved. It is ineffective in men in whom both NVBs have been excised, and it is also ineffective in older men in whom only one NVB has been preserved. Sildenafil appears ineffective in the first 9 months after prostatectomy. PMID- 11018621 TI - Asian sildenafil efficacy and safety study (ASSESS-1): a double-blind, placebo controlled, flexible-dose study of oral sildenafil in Malaysian, Singaporean, and Filipino men with erectile dysfunction. The Assess-1 Study Group. AB - OBJECTIVES: To evaluate the efficacy, safety, and tolerability of oral sildenafil in Asian men with erectile dysfunction of various causes (organic, psychogenic, or mixed) and of more than 6 months' duration. METHODS: In this double-blind, parallel-group trial conducted at eight centers in Malaysia, the Philippines, and Singapore, 254 men, 26 to 78 years old, were randomized to 12 weeks of sildenafil or placebo taken as needed 1 hour before anticipated sexual activity. Initially, the sildenafil (n = 127) or matching placebo (n = 127) dose was 50 mg but could be increased to 100 mg or decreased to 25 mg because of a lack of efficacy or intolerance, respectively. Efficacy was assessed by the 15-question International Index of Erectile Function, patients' event logs of sexual activity, and a global efficacy question about erections. RESULTS: The two primary efficacy variables relating to achievement and maintenance of an erection sufficient for sexual intercourse, as assessed by the mean scores for International Index of Erectile Function question 3 (4.22 versus 2.59) and question 4 (4.15 versus 2.41), were both significantly higher with sildenafil than with placebo (P <0.0001). In addition, the five separate International Index of Erectile Function domains of sexual function, the percentage of successful intercourse attempts, and the global efficacy assessment of erections revealed significantly greater treatment effects in favor of sildenafil (P <0.0001 versus placebo for all variables). Treatment-related adverse events occurred in 22.8% of patients who received sildenafil and in 10.2% of those who received placebo. CONCLUSIONS: Sildenafil is an effective and well-tolerated treatment for Asian men with erectile dysfunction of broad-spectrum etiology. PMID- 11018622 TI - Effect of an alpha-melanocyte stimulating hormone analog on penile erection and sexual desire in men with organic erectile dysfunction. AB - OBJECTIVES: To assess the safety, erectogenic properties, and effect on sexual desire of Melanotan II, a synthetic melanotropic initiator of erection, in men with erectile dysfunction and organic risk factors. METHODS: Ten subjects were enrolled in a double-blind, placebo-controlled, crossover study. Melanotan II (0.025 mg/kg) and vehicle were each administered twice by subcutaneous injection; real-time RigiScan monitoring and a visual analog were used to quantify the erections during a 6-hour period. The level of sexual desire and side effects were recorded with a questionnaire. RESULTS: Melanotan II initiated subjectively reported erections in 12 of 19 injections versus only 1 of 21 doses of placebo. The mean rigidity score of the responders was 6.9 on a scale of 0 to 10. The mean duration of tip rigidity greater than 80% was 45.3 minutes with Melanotan II versus 1.9 for placebo (P = 0.047). The level of sexual desire after injection was significantly higher after Melanotan II administration than after placebo. Nausea and stretching/yawning occurred more frequently with Melanotan II, and 4 of 19 injections were associated with severe nausea. CONCLUSIONS: The erectogenic properties of Melanotan II are not limited to cases of psychogenic erectile dysfunction; men with a variety of organic risk factors developed penile erections. The finding of increased sexual desire warrants further investigation of centrally acting agents on disorders of sexual desire. PMID- 11018623 TI - Three-year outcome of a progressive treatment program for erectile dysfunction with intracavernous injections of vasoactive drugs. AB - OBJECTIVES: To perform a comparative evaluation and follow-up of patients with erectile dysfunction (ED) who were treated with intracavernous injection of vasoactive drugs, starting with simple drugs and advancing to complex combinations. METHODS: The study included 625 patients, 26 to 85 years old, with ED. Four intracavernous injection protocols were used: protocol 1, papaverine plus phentolamine; protocol 2, prostaglandin E(1); protocol 3, papaverine, phentolamine, and prostaglandin E(1); and protocol 4, atropine sulfate, papaverine, phentolamine, and prostaglandin E(1). A positive response was defined as an erection sufficient for penetration. Patients for whom the basic protocol failed were successively switched to the more advanced protocols until a positive response was achieved. RESULTS: A positive response was achieved by 415 (66.4%) of the 625 patients given protocol 1; 75 (36%) of the remaining 210 patients given protocol 2; 98 (72.6%) of the 135 patients given protocol 3; and 22 (59.5%) of 37 patients given protocol 4. All four protocols failed in only 15 patients (2. 4%). At the 3-year follow-up visit (n = 610), 349 had achieved coitus, 65 (10.6%) without an injection and 202 (33.1%) with an injection. Eighty-two patients sometimes performed coitus without an injection. Sixty-three patients (10.3%) abandoned the program because of marital or health problems; 198 asked to be switched to Viagra during the follow-up period, and 120 returned to the program. CONCLUSIONS: Overall, our progressive treatment yielded a high positive response rate (97.6%), with 57.2% achieving successful coitus on follow-up. The main advantage of the program is that it spares patients who are responsive earlier from using more complex, painful, and costly drugs. PMID- 11018625 TI - Editorial comment. Wide interpretation of PUV. PMID- 11018624 TI - Outcomes of primary valve ablation versus urinary tract diversion in patients with posterior urethral valves. AB - OBJECTIVES: Although valve ablation is the treatment of choice for patients with posterior urethral valves, debate continues as to the role of urinary diversion. We sought to retrospectively compare the clinical and radiologic outcomes between valve ablation and urinary diversion for patients with posterior urethral valves. METHODS: We retrospectively reviewed the records of 50 consecutive patients with posterior urethral valves since January 1995. On the basis of the initial renal function and radiologic findings, patients were divided into three groups: group 1, normal renal function and radiologically normal upper tracts; group 2, normal renal function with hydronephrosis and/or reflux; and group 3, azotemia with hydronephrosis or reflux. RESULTS: All 22 patients in group 1 were treated with valve ablation. After a mean follow-up of 32 months, these children had normal renal function and no evidence of upper tract deterioration. All 13 patients in group 2 were also treated with valve ablation. The radiologic abnormalities (hydronephrosis, reflux) resolved in 50% of cases, with an average follow-up of 28 months. Of the 15 patients in group 3, 7 underwent valve ablation and 8 underwent urinary diversion. Urinary diversion was performed in patients with renal deterioration and severe hydronephrosis and/or high-grade reflux. Renal function returned to normal in all patients who underwent valve ablation except one; renal function returned to normal in only 3 of 8 patients who underwent urinary diversion. Radiologically, the severity of the hydronephrosis and reflux was downgraded in patients who underwent valve ablation but not in the diverted group. CONCLUSIONS: Valve ablation is the mainstay of treatment for patients with posterior urethral valves. Prenatal and postnatal factors, such as renal dysplasia and urinary tract infection, respectively, rather than the posterior valve treatment dictate the long-term renal and radiologic outcomes. PMID- 11018626 TI - Reply by the authors PMID- 11018627 TI - New method to improve treatment outcomes for radical prostatectomy. AB - Modifications in radical retropubic prostatectomy that use the deep dorsal vein ligator and either anterior urethropexy or puboprostatic ligament preservation were developed to reduce blood loss and improve continence results. Use of the deep dorsal vein ligator to secure the deep dorsal vein complex also improves visualization of the urethra for transection and the neurovascular bundle for nerve sparing and thereby may assist in laparoscopic radical retropubic prostatectomy. PMID- 11018628 TI - Feasibility of transperineal prostate biopsy under interventional magnetic resonance guidance. AB - A patient, who was not suited for ultrasound-guided biopsy, was biopsied in an interventional magnetic resonance unit at Brigham and Women's Hospital. Real-time magnetic resonance imaging provided verification of placement before tissue samples were taken. This technique successfully sampled tissue from the prostate and led to a diagnosis of cancer. PMID- 11018629 TI - Controversies in the management of ectopic ureteroceles. PMID- 11018630 TI - Ulcerative balanoposthitis of the foreskin as a manifestation of chronic lymphocytic leukemia: case report and review of the literature. AB - Ulcerative lesions of the penis have many possible etiologies, including infectious, neoplastic, traumatic, drug-induced, and autoimmune. Although the most frequent neoplasm presenting as an ulcerative penile lesion is squamous cell carcinoma, it may rarely be a manifestation of other malignancies, including those of hematolymphoid origin. We report a case of ulcerative balanoposthitis as a manifestation of chronic lymphocytic leukemia. Chronic lymphocytic leukemia and other hematolymphoid malignancies should be considered in the large differential diagnosis of nonhealing penile ulcers. PMID- 11018631 TI - Peripheral nerve injury after brief lithotomy for transurethral collagen injection. AB - Two patients with prior prostate surgery sustained peripheral nerve injuries after transurethral collagen injection for the treatment of urinary incontinence. In the first patient, brief lithotomy positioning caused a gluteal compartment syndrome and sciatic neuropathy. In the second patient, obturator neuropathy was due to leakage of collagen along the course of the obturator nerve. This is the first report of peripheral nerve injury in patients undergoing transurethral collagen injection. PMID- 11018632 TI - Invasive group A streptococcus infection of the scrotum and streptococcal toxic shock syndrome. AB - We report a case of invasive group A streptococcus infection of the scrotum that presented as epididymoorchitis and rapidly progressed to streptococcal toxic shock syndrome. The presentation, pathophysiology, and management of invasive group A streptococcus and streptococcal toxic shock syndrome are reviewed. Rapid recognition is necessary to avoid the significant morbidity and mortality associated with these invasive infections. PMID- 11018633 TI - Leiomyosarcoma of the main renal artery treated by laparoscopic radical nephrectomy. AB - Renal leiomyosarcomas are rare mesenchymal sarcomas. Although such tumors arising from the renal vein or kidney have been previously reported, we present the first case of a leiomyosarcoma arising from the main renal artery managed by laparoscopic radical nephrectomy. PMID- 11018635 TI - Plexiform neurofibroma of the penis in a child. AB - We report a case of a child with the stigmata of von Recklinghausen's disease and a plexiform neurofibroma arising from the penile shaft. PMID- 11018634 TI - Radioimmunoguided surgery using indium-111 capromab pendetide (PROSTASCINT) to diagnose supraclavicular metastasis from prostate cancer. AB - A 59-year-old man presented with a Gleason score of 4 + 4 = 8 prostate cancer and with multiple bilateral pelvic nodes involved at open pelvic lymph node dissection. On indium-111 capromab pendetide (ProstaScint) scan, there was increased tracer deposition in the prostate, the mesenteric nodes, the right pulmonary hilum, and the left supraclavicular fossa. The ProstaScint injection was repeated, and the gamma probe was used to localize tissue that accumulated radiotracer. Two nodes were excised, one that exhibited increased uptake and one that did not. The radioactive lymph node contained metastatic prostate cancer. No malignancy was found in the second node. PMID- 11018636 TI - Fibrous pseudotumor of tunica vaginalis and epididymis. AB - Paratesticular fibrous pseudotumors are rare, and less than 10% of them affect the epididymis. We report a case of testicular trauma that progressed to a painless palpable tumor in the right hemiscrotum with increased local volume. Orchiectomy followed by anatomic-pathologic examination revealed a paratesticular fibrous pseudotumor. The best treatment is scrotal exploration and frozen biopsy. On confirmation of the diagnosis, only the tumor and the tunica vaginalis are resected. Should the benign nature not be possible to determine, orchiectomy is the procedure of choice. We also reviewed published reports for this possibility in the differential diagnosis of testicular masses. PMID- 11018637 TI - Expression of cyclooxygenase-1 and cyclooxygenase-2 in the human prostate. AB - OBJECTIVES: To determine the cell-specific expression of the two major isoforms of cyclooxygenase (COX-1 and COX-2) in human noncancerous and cancerous prostatic tissues. METHODS: Thirty-one specimens of prostate carcinoma (CaP) and 10 specimens of benign prostatic hyperplasia (BPH) were stained with mouse antihuman COX-1 and COX-2 monoclonal antibodies. The stained specimens were analyzed both descriptively and in a semiquantitative manner by assigning an immunoreactive intensity score (0 to 4). The averaged results were compared for different histologic tissue types, including luminal and basal epithelium of BPH, the peripheral zone, high-grade prostatic intraepithelial neoplasia (PIN), and CaP of varying Gleason grades. RESULTS: COX-1 expression in noncancerous prostatic tissue was seen predominantly in the basal epithelial cells of BPH (90% positive staining). COX-1 expression was minimal in noncancerous luminal epithelial cells (0% to 10%) but was upregulated in CaP (63% of CaP specimens). Strong COX-2 expression was demonstrated in the smooth muscle cells of the prostate. COX-2 was also expressed in the basal epithelial cells (60% BPH, 94% peripheral zone, 75% PIN). Luminal epithelial cells derived from BPH, the peripheral zone, and PIN expressed COX-2 in 0%, 26%, and 86% of samples, respectively. COX-2 expression in CaP was intense and uniform, with 87% of samples demonstrating immunoreactivity. CONCLUSIONS: The results of the present study indicate that expression of both COX-1 and COX-2 in human CaP is increased. COX-2 expression is also increased in the basal and luminal epithelial cells of PIN. These data indicate that COX-1 and COX-2 (and/or their prostaglandin products) may play a role in the malignant transformation of the prostate. PMID- 11018638 TI - Laparoscopic renal ablation: an in vitro comparison of currently available electrical tissue morcellators. AB - OBJECTIVES: Morcellation with the Cook high-speed electrical laparoscopic (HSEL) morcellator in an impermeable nylon/plastic sack (LapSac) has remained unchanged since its inception nearly one decade ago. Sack deployment and specimen entrapment remain relatively difficult, and morcellation with this device is expensive and relatively slow. As such, in an effort to facilitate specimen entrapment and morcellation, we adapted two currently available electrical morcellators (the Steiner gynecologic morcellator and the electrical prostate morcellator [EPM]) for renal morcellation and compared them with the HSEL morcellator. METHODS: All morcellation was performed through a simulated abdominal wall under direct laparoscopic vision. Ten porcine kidneys were ablated with each of the following techniques: HSEL morcellation in a LapSac; HSEL morcellation in a fluid-filled LapSac; Steiner morcellation in an insufflated Endocatch sack; and EPM morcellation in a fluid-filled Endocatch sack. A modified laparoscopic trocar was constructed and used for the Steiner and EPM morcellation. The time to complete morcellation, morcellation product size, and entrapment sack integrity were evaluated for each technique. Cost data for each morcellator are also presented. RESULTS: The mean morcellation time for the Steiner, HSEL dry, HSEL wet, and EPM morcellation was 6.0, 15.9, 14.7, and 26.0 minutes, respectively. The mean fragment size for these morcellators was 2.97, 0.65, 0.62, and 0.013 g, respectively. A single entrapment sack perforation was documented in a LapSac during routine HSEL morcellation. CONCLUSIONS: Renal morcellation with all three morcellators is feasible. The Steiner morcellator combined with an Endocatch resulted in more rapid morcellation and larger morcellation products. PMID- 11018639 TI - Time-resolved fluorescence imaging for specific and quantitative immunodetection of human kallikrein 2 and prostate-specific antigen in prostatic tissue sections. AB - OBJECTIVES: To design protocols for specific and quantitative immunohistochemical detection of human kallikrein 2 (hK2) using lanthanide chelate-labeled monoclonal antibodies (Mabs) and time-resolved fluorescence imaging. METHODS: Anti-prostate specific antigen (PSA) Mabs were tested in microtiterplate assays for their ability to prevent PSA from cross-reacting with the anti-hK2 Mab 6H10. Europium labeled 6H10 and terbium-labeled anti-PSA Mab 2E9, selected as the best blocker antibody, were used for dual-label immunodetection in routinely fixed benign (n = 7) and malignant (n = 5) prostate specimens. The amounts of IgG bound in tissue were calculated from drops containing known Mab concentrations. RESULTS: The use of anti-PSA Mab 2E9 for blocking diminished the cross-reaction from 5% to 0.3%. In the analyzed tissues, there was considerable variation in staining intensity for both proteins; PSA signals varied from 0.1 to 36.6 times that of hK2, with on average 10-fold more bound anti-PSA Mab than anti-hK2 Mab. In malignant tissue, the amounts of bound IgGs were lower and more variable than in benign tissue using both the anti-PSA Mab and the anti-hK2 Mab. The variation in signal intensities for PSA and hK2 correlated significantly in benign tissue (P >0.05), but not in benign hyperplastic and malignant specimens (P <0.05). CONCLUSIONS: Quantification of two lanthanide chelate-labeled antibodies bound in the same tissue section enabled comparison of PSA and hK2 content in individual cells. The average cellular content of hK2 relative to that of PSA was consistent with previous mRNA studies. The time-resolved fluorescence imaging-based quantification method has universal applicability in fixed tissue specimens. PMID- 11018640 TI - Cyclin-dependent kinase inhibitor P27(KIP1) is expressed preferentially in early stages of urothelial carcinoma. AB - OBJECTIVES: To evaluate the expression of p27(KIP1) and p21(CIP1) and the prognostic values of both markers in urothelial carcinoma. The expression of the cyclin-dependent kinase inhibitor p27(KIP1) characterizes early-stage and well differentiated carcinomas of the colon, breast, and prostate and is associated with an improved prognosis. In urothelial carcinoma, its expression has not been as well investigated. Another cyclin-dependent kinase inhibitor, p21(CIP1), is expressed in early-stage bladder tumors, but published data on its prognostic value are contradictory. METHODS: Expression of p27(KIP1) and p21(CIP1) was analyzed by immunohistochemistry in 114 urothelial carcinoma specimens from 77 patients. The Ki67 index was determined as an indicator of cell proliferation. The expression of the markers was correlated with tumor recurrence and progression during an average follow-up period of 3.9 years. RESULTS: Expression of p27(KIP1) was significantly more frequent in superficial than in muscle invasive tumors (chi-square test, P = 0.012; Fisher's exact test, P = 0.014). Although similar overall, the expression pattern of p21(CIP1) did not match on a tumor-by-tumor basis. No correlation was seen with the Ki67 index. Patients with tumors displaying strong positive staining for p27(KIP1) or p21(CIP1) had fewer recurrences and progression events, but the difference was not statistically significant. Instead, a Ki67 index of less than 10% was significantly (P = 0.0335) related to a lack of recurrence. CONCLUSIONS: Neither p27(KIP1) nor p21(CIP1) appear to be good predictors of tumor progression in urothelial carcinoma, even though their expression is strongly decreased in muscle-invasive tumors. PMID- 11018641 TI - Simultaneous recording of mechanical and intracellular electrical activity in guinea-pig urinary bladder smooth muscle: a comparison with human detrusor contraction. AB - OBJECTIVES: To study the relationship between the electrical and mechanical activity of guinea-pig detrusor muscle and compare this with the results obtained in human tissue. The guinea pig is the most used model to study the electrical basis of human bladder contraction. METHODS: We simultaneously recorded the mechanical and intracellular electrical activity in muscle strips. To study the effect of tissue movement on the membrane potential, the medium was made hypertonic with sucrose. Carbachol and KCl were applied to the bath to induce contractions. RESULTS: Carbachol resulted in a force response without a consistent change in the membrane electrical activity. KCl induced depolarization of the membrane associated with force development. Sucrose in the medium greatly impaired the ability to contract, without affecting the electrical activity. Compared with recordings in normal Krebs' solution, the resting membrane potential was not altered. In both media, spike-shaped potentials with variable amplitudes and shapes were recorded. These events were minimally affected by sucrose. CONCLUSIONS: In contrast to the response to KCl, the overall mechanical response of guinea-pig detrusor strips to muscarinic receptor stimulation did not correlate with the electrical activity of a single cell. Sucrose had only a minimal effect on the electrical activity, demonstrating that the electrical responses we measured were not affected by movement. Our intracellular recordings in guinea-pig tissue differed from the results obtained by other groups, but show great resemblance with those we recorded in human urinary bladder smooth muscle strips. PMID- 11018642 TI - Mannitol facilitates rabbit urinary bladder recovery from overdistension injury. AB - OBJECTIVES: To investigate the existence and functional significance of the enhanced lipid peroxidation in bladder overdistension injury and to explore the effect of mannitol, a free radical scavenger. METHODS: Overdistension of rabbit bladders was induced and maintained for 3 hours by infusing normal saline into the bladder while keeping the intravesical pressure at 30 cm H(2)O. The bladders were then emptied and decompressed. Intravenous 20% mannitol was initiated 5 minutes before decompressing the overdistension. Detrusor tissue was obtained from the following groups: control, at the end of the overdistension period, and 30 minutes, 2 hours, and 7 days after decompressing the bladder. The tissue level of adenosine triphosphate (ATP) and phosphocreatine (PCr) and the lipid peroxidation product malondialdehyde (MDA) was assayed. Detrusor contractility was assessed by the response of the detrusor strips to KCl and bethanechol. RESULTS: Decompressing the overdistended bladder led to a period of enhanced lipid peroxidation with an increase of MDA content from 225 to 384 pmol/mg protein 30 minutes after the decompression. Two hours later, the MDA content had recovered to the normal level. Mannitol abolished this period of enhanced lipid peroxidation. Overdistension impaired detrusor contractility and reduced the content of PCr (from 24.1 to 10.8 nmol/mg protein) and ATP (from 9.6 to 4.6 nmol/mg protein). Both detrusor contractility and the content of PCr and ATP further decreased 30 minutes after the decompression (PCr 5.4 nmol/mg, ATP 2.8 nmol/mg). They had recovered, but not fully, 7 days later. Mannitol prevented the further decrease in detrusor contractility and in the content of PCr and ATP during the initial decompression period (30 minutes after the decompression). In addition, the mannitol-treated group had quicker recovery in PCr and ATP levels, which returned to normal 7 days later. CONCLUSIONS: Decompressing an overdistended bladder leads to enhanced lipid peroxidation, which is associated with an additionally decreased energetic metabolism and a more impaired contractile function. Mannitol effectively prevents enhanced lipid peroxidation and facilitates functional recovery. These results show that reactive oxygen species play a significant role in bladder overdistension injury. PMID- 11018643 TI - "Latent" carcinoma of the prostate: a medical misnomer? PMID- 11018644 TI - Purification of cytochrome b-561 from bean hypocotyls plasma membrane. Evidence for the presence of two heme centers. AB - The high potential, ascorbate-reducible b-type cytochrome of plant plasma membranes, named cytochrome b-561, has been purified to homogeneity from etiolated bean hypocotyls. The pure protein migrated in denaturing electrophoresis as a broad band of approximately 55 kDa, and was found to be glycosylated. Optical redox titrations of partially purified cytochrome b-561 indicated that it contains two hemes with similar spectral features, but distinct midpoint redox potentials (E(m7)+135 mV and +206 mV, respectively). The presence of two heme centers in cytochrome b-561 is consistent with its role in electron transfer across plant plasma membranes. PMID- 11018645 TI - Efficient in vivo gene delivery by the negatively charged complexes of cationic liposomes and plasmid DNA. AB - We examined changes in zeta potential (the surface charge density, zeta) of the complexes of liposome (nmol)/DNA (microg) (L/D) formed in water at three different ratios (L/D=1, 10 and 20) by changing the ionic strength or pH to find an optimum formulation for in vivo gene delivery. At high DNA concentrations, zeta of the complexes formed in water at L/D=10 was significantly lowered by adding NaCl (zeta=+8.44+/-3.1 to -27.6+/-3.5 mV) or increasing pH from 5 (zeta=+15.3+/-1.0) to 9 (zeta=-22.5+/-2.5 mV). However, the positively charged complexes formed at L/D=20 (zeta=+6.2+/-3.5 mV) became negative as NaCl was added at alkaline pH as observed in medium (zeta=-19.7+/-9.9 mV). Thus, the complexes formed in water under the optimum condition were stable and largely negatively charged at L/D=1 (zeta=-58.1+/-3.9 mV), unstable and slightly positively charged at L/D=10 (zeta=+8.44+/-3.7 mV), and unstable and largely positively charged at L/D=20 (zeta=+24.3+/-3.6 mV). The negatively charged complexes efficiently delivered DNA into both solid and ascitic tumor cells. However, the positively charged complexes were very poor in delivering DNA into solid tumors, yet were efficient in delivering DNA into ascitic tumors grown in the peritoneum regardless of complex size. This slightly lower gene transfer efficiency of the negatively charged complexes can be as efficient as the positively charged ones when an injection is repeated (at least two injections), which is the most common case for therapy regimes. The results indicate that optimum in vivo lipofection may depend on the site of tumor growth. PMID- 11018646 TI - Zeta potential of transfection complexes formed in serum-free medium can predict in vitro gene transfer efficiency of transfection reagent. AB - We have tested the zeta potential (zeta, the surface charge density) of transfection complexes formed in serum-free medium as a rapid and reliable technique for screening transfection efficiency of a new reagent or formulation. The complexes of CAT plasmid DNA (1 microgram) and DC-chol/DOPE liposomes (3-20 nmol) were largely negatively charged (zeta=-15 to -21 mV), which became neutral or positive as 0.5 microgram or a higher amount of poly-L-lysine (PLL, MW 29300 or MW 204000) was added (-3.16+/-3.47 to +6.04+/-2.23 mV). However, the complexes of CAT plasmid DNA (1 microgram) and PLL MW 29300 (0.5 microgram or higher) were neutral or positively charged (-3.22+/-2.3 to +6.55+/-0.64 mV), which remained the same as 6.6 nmol of the liposomes was added. The complexes formed between two positively charged compounds, PLL MW 29300 (0.5 microgram) and the liposomes (3 20 nmol), were as closely positively charged as DNA/PLL or DNA/liposomes/PLL complexes (+3.31+/-0.41 to 7.16+/-1.0 mV). These results indicate that PLL determined the overall charge of the DNA/liposome/PLL ternary complexes. The complexes formed with histone (0.75 microgram or higher) were also positively charged, whose transfection activity was as high as PLL MW 29300. However, the complexes formed with protamine or PLL MW 2400 remained negatively charged. These observations are in good agreement with the transfection activity of the formulation containing each polycationic polymer. The presence of PLL MW 29300 did not change the hydrodynamic diameter of DNA/liposome/PLL complexes (d(H)=275 312 nm). The complexes made of different sizes of PLL (MW 2400 and 204000) also did not significantly change their size. This suggests that DNA condensation may not be critical. Therefore, zeta of the transfection complex can predict the transfection efficiency of a new formulation or reagent. PMID- 11018647 TI - Involvement of membrane surface charge in thermal stability of the rat ovarian LH/hCG receptor. AB - Analysis of fluorescence of membrane-bound 8-anilino-1-naphthalene sulfonate and monodansylcadaverine probes revealed that a negative membrane surface charge derived from free fatty acids (FFA) resulted in destabilization of structure functional properties of the rat ovarian LH/hCG receptor. Removal of FFA from rat luteal and porcine ovarian granulosa cells by BSA increased gonadotropin responsiveness of cells in cAMP formation. PMID- 11018648 TI - Characterization of a new class of DNA delivery complexes formed by the local anesthetic bupivacaine. AB - Bupivacaine, a local anesthetic and cationic amphiphile, forms stable liposomal like structures upon direct mixing with plasmid DNA in aqueous solutions. These structures are on the order of 50-70 nm as determined by scanning electron microscopy, and are homogeneous populations as analyzed by density gradient centrifugation. The DNA within these structures is protected from nuclease degradation and UV-induced damage in vitro. Bupivacaine:DNA complexes have a negative zeta potential (surface charge), homogeneous nature, and an ability to rapidly assemble in aqueous solutions. Bupivacaine:DNA complexes, as well as similar complexes of DNA with other local anesthetics, have the potential to be a novel class of DNA delivery agents for gene therapy and DNA vaccines. PMID- 11018649 TI - Doxorubicin entrapped within liposome-associated antigens results in a selective inhibition of the antibody response to the linked antigen. AB - The generation of an immune response can dramatically alter the circulation lifetime of a targeted liposome, particularly when the response is generated against the surface-coupled ligand. Following repeated administrations, rapid elimination of the carrier system is observed, thereby limiting potential applications for targeted liposomes in a therapeutic setting. In this study, we have investigated whether the encapsulation of a toxic drug within the carrier could prevent an immune response against a surface-bound protein. Liposome clearance and humoral immune response were monitored throughout multiple administrations of liposomes containing doxorubicin with surface-conjugated ovalbumin. The results show that low doses of encapsulated doxorubicin can prevent humoral immunity against repeated administration of liposomes conjugated with ovalbumin. The immunosuppressive effect was specific for the ovalbumin coupled to the liposome surface. This selective suppression of immunity against a surface conjugated protein could prove advantageous for safe repeated administration of protein containing liposomal systems. PMID- 11018650 TI - Estimations of lipid bilayer geometry in fluid lamellar phases. AB - The excess water bilayer thickness, d(l,0), and molecular area, A(0), of lipid amphiphiles in the fluid lamellar phases of dioleoylphosphatidylcholine (DOPC) and dipalmitoleoylphosphatidylcholine (DPolPC) have been estimated between 15 and 50 degrees C and for dimyristoylphosphatidylcholine (DMPC) between 25 and 50 degrees C. These determinations have been made from X-ray measurements on samples of known water composition. With respect to temperature, T, d(l,0) and A(0) are well fitted to a linear equation. We find d(l,0) (A)=(35.68+/-0.02)-(0.0333+/ 0.0006)T (degrees C) and A(0) (A(2))=(70.97+/-0.05)+(0.136+/-0.001)T (degrees C) for DOPC, d(l,0) (A)=(35.2+/-0.1)-(0.068+/-0.003)T (degrees C) and A(0) (A(2))=(59.7+/-0.2)+(0.210+/-0.006)T (degrees C) for DMPC, and d(l,0) (A)=(34.54+/-0.03)-(0.0531+/-0.0009)T (degrees C) and A(0) (A(2))=(67.12+/ 0.09)+(0.173+/-0.003)T (degrees C) for DPolPC. The accuracy of these estimates depends largely on how accurately the excess water point is determined. Ideally, reliable X-ray and compositional data will be available around the excess water and it may be found by simple inspection, but this is the exception rather than the rule, since samples close to water excess normally sequester sizeable amounts of water in defects, which lead to an underestimate of d(l,0). and overestimate of A(0). In this paper, we report a methodology for identifying and removing such data points and fitting the remaining data in order to determine the excess water point. PMID- 11018651 TI - Characterization of the ATP transporter in the reconstituted rough endoplasmic reticulum proteoliposomes. AB - Adenosine triphosphate (ATP) transporter from rat liver rough endoplasmic reticulum (RER) was solubilized and reconstituted into phosphatidylcholine liposomes. The RER proteoliposomes, resulting from optimizing some reconstitution parameters, had an apparent K(m) value of 1.5 microM and a V(max) of 286 pmol min(-1) (mg protein)(-1) and showed higher affinity for ATP and a lower V(max) value than intact RER (K(m) of 6.5 microM and V(max) of 1 nmol). ATP transport was time- and temperature-dependent, inhibited by 4, 4'-diisothiocyanostilbene 2,2'-disulfonic acid, which is known as an inhibitor of anion transporters including ATP transporter, but was not affected by atractyloside, a specific inhibitor of mitochondrial ADP/ATP carrier. The internal and external effects of various nucleotides on the ATP transport were examined. ATP transport was cis inhibited strongly by ADP and weakly by AMP. ADP-preloaded RER proteoliposomes showed a specific increase of ATP transport activity while AMP-preloaded RER proteoliposomes did not show the enhanced overshoot peak in the ATP uptake plot. These results demonstrate the ADP/ATP antiport mechanism of ATP transport in rat liver RER. PMID- 11018652 TI - Dual role of ATP in supporting volume-regulated chloride channels in mouse fibroblasts. AB - The effects of inhibitors of protein tyrosine kinases (PTKs) on the Cl(-) current (I(Cl(vol))) through volume-regulated anion/chloride (VRAC) channels whilst manipulating cellular ATP have been studied in mouse fibroblasts using the whole cell patch clamp technique. Removal of ATP from the pipette-filling solution prevented activation of the current during osmotic cell swelling and when the volume of patched cells was increased by the application of positive pressure through the patch pipette to achieve rates exceeding 100%/min. Equimolar substitution of ATP in the pipette solution with its non-hydrolyzable analogs, adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS) or adenylyl-(beta,gamma methylene)-diphosphonate (AMP-PCP), not only supported activation of the current but also maintained its amplitude. The PTK inhibitors, tyrphostins A25, B46, 3 amino-2,4-dicyano-5-(4-hydroxyphenyl)penta-2,4-dienonitrile++ + and genistein (all at 100 microM), inhibited I(Cl(vol)) in a time-dependent manner. Tyrphostin A1, which does not inhibit PTK activity, did not affect the current amplitude. The PTK inhibitors also inhibited I(Cl(vol)) under conditions where ATP in the pipette was substituted with ATPgammaS or AMP-PCP. We conclude that in mouse fibroblasts ATP has a dual role in the regulation of the current: it is required for protein phosphorylation to keep VRAC channels operational and, through non hydrolytic binding, determines the magnitude of I(Cl(vol)). We also suggest that tyrosine-specific protein kinases and phosphatases exhibit an interdependent involvement in the regulation of VRAC channels. PMID- 11018653 TI - Effect of ethylene oxide and propylene oxide block copolymers on the permeability of bilayer lipid membranes to small solutes including doxorubicin. AB - The effects of ethylene oxide and propylene oxide block copolymers (pluronics) on the permeability of several weak acids and bases through bilayer lipid membranes have been studied by the methods of monitoring (1) pH shifts near planar bilayers, (2) doxorubicin fluorescence quenching inside liposomes, and (3) current transients in the presence of hydrophobic anions. It has been shown that pluronics facilitate the permeation of comparatively large molecules (such as 2-n undecylmalonic acid and doxorubicin) across lipid bilayers, while the permeation of small solutes (such as ammonium and acetic acid) remains unaffected. Pluronics also accelerate the translocation of large hydrophobic anions (tetraphenylborate). The effect of pluronics correlates with the content of propylene oxide units: it is enhanced when the portion of polypropylene oxide block in the copolymer is increased. The action of the pluronic on lipid membrane permeability differs from the effect of the conventional detergent Triton X-100, which does not affect doxorubicin transport if added at concentrations similar to those used for pluronics. It has been proposed that pluronics accelerate the processes of solute diffusion within lipid bilayers (in a structure-dependent manner) rather than influencing the rate of solute adsorption/desorption on the membrane surface. We suppose that the effect of pluronics on doxorubicin permeation across lipid bilayers along with the known effect on the multidrug resistance protein determines its influence on the therapeutic activity of anthracycline drugs. PMID- 11018654 TI - Effect of influenza hemagglutinin fusion peptide on lamellar/inverted phase transitions in dipalmitoleoylphosphatidylethanolamine: implications for membrane fusion mechanisms. AB - Low mole fractions of viral fusion peptides induce inverted cubic (Q(II)) phases in dipalmitoleoylphosphatidylethanolamine (DiPoPE), a lipid with unsaturated acyl chains that normally forms inverted hexagonal phase (H(II)) above 43 degrees C. The ability to form a Q(II) phase is relevant to the study of membrane fusion: fusion occurs in liposomal systems under conditions where Q(II) phase precursors form, and fusion may be an obligatory step in the lamellar (L(alpha))/Q(II) phase transition. We used X-ray diffraction and time-resolved cryoelectron microscopy (TRC-TEM) to study the effects of the influenza hemagglutinin fusion peptide on the phase behavior and structure of DiPoPE. X-ray diffraction data show that at concentrations of 3-7 mol%, the fusion peptide (FP) induces formation of a Q(II) phase in preference to the H(II) phase. TRC-TEM data show that the FP acts at early stages in the phase transition (i.e. within seconds): at 2-7 mol%, FP decreases or inhibits formation of the L(alpha)/H(II) intermediate morphology observed via TRC-TEM in pure DiPoPE at the same temperature. Our X-ray diffraction data imply that FP either does not affect, or slightly increases, the spontaneous curvature of the host lipid (i.e. either does not affect or tends to destabilize inverted phases, respectively). FP may act in part by affecting the relative stability of two intermediate structures in the phase transition mechanism, as suggested previously. These results indicate a new way in which hydrophobic sequences of membrane proteins may be fusogenic. PMID- 11018655 TI - Genetic characterization of the (534)DPPR motif of the yeast plasma membrane H(+) ATPase. AB - The highly conserved motif +(534)DPPR of Saccharomyces cerevisiae H(+)-ATPase, located in the putative ATP binding site, has been mutagenized and the resulting 23 mutant genes conditionally expressed in secretory vesicles. Fourteen mutant ATPases (D534A, D534V, D534L, D534N, D534G, D534T, P535A, P535V, P535L, P535G, P535T, P535E, P535K and R537T) failed to reach the secretory vesicles. Of these mutants, nine (D534N, D534T, P535A, P535V, P535L, P535G, P535T, P535E and P535K) were not detected in total cellular membranes, and five (D534A, D534V, D534G, D534L and R537T) were retained at the endoplasmic reticulum and exhibited a dominant lethal phenotype. The remaining mutants (D534E, R537A, R537V, R537L, R537N, R537G, R537E, R537K and R537H) reached the secretory vesicles at levels similar to that of the wild type. Of these, six (R537A, R537V, R537L, R537N, R537G, and R537E) showed severely decreased ATPase activity compared to the wild type enzyme, and three (D534E, R537K and R537H) rendered an enzyme with an altered K(m) for ATP. PMID- 11018656 TI - Ouabain-insensitive Na(+)-ATPase activity is an effector protein for cAMP regulation in basolateral membranes of the proximal tubule. AB - This study describes the modulation of the ouabain-insensitive Na(+)-ATPase activity from proximal tubule basolateral membranes by cAMP. An increase in dibutyryl-cAMP (d-cAMP) concentration from 10(-8) to 5x10(-5) M stimulates the ouabain-insensitive Na(+)-ATPase activity. The ATPase activity increases from 6.0+/-0.4 to 10.1+/-0.7 nmol Pi mg(-1) min(-1), in the absence and presence of 5x10(-6) M d-cAMP, respectively. Similarly, the addition of cholera toxin (CTX), forskolin (FSK) or guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) also increases the Na(+)-ATPase activity in a dose-dependent manner, with maximal effect at 10( 8) M, 10(-6) M and 10(-7) M, respectively. The effect of 10(-8) M CTX is not additive to the effect of GTPgammaS, and is completely abolished by 200 microM guanosine 5'-O-(2-thiodiphosphate). The stimulatory effects of CTX and FSK on the Na(+)-ATPase activity are accompanied by an increase in cAMP formation by the basolateral membranes of the proximal tubule cells. Furthermore, 10(-8) M protein kinase A peptide inhibitor (PKAi) completely abolishes the stimulatory effect of 5x10(-6) M d-cAMP or 10(-4) M FSK on the Na(+)-ATPase activity. Incubation of the basolateral membranes with [gamma-(32)P]ATP in the presence of d-cAMP or FSK increases the global hydroxylamine-resistant phosphorylation and especially promotes an increase in phosphorylation of protein bands of approximately 100 and 200 kDa. This stimulation is not seen when 10(-8) M PKAi is added simultaneously. Taken together these data suggest that activation of a cAMP/PKA pathway modulates the Na(+)-ATPase activity in isolated basolateral membranes of the proximal tubule. PMID- 11018657 TI - Partly folded states of bovine carbonic anhydrase interact with zwitterionic and anionic lipid membranes. AB - The acidic, partly folded states of bovine carbonic anhydrase II (BCAII) were used as an experimental system to study the interactions of partly denatured proteins with lipid membranes. The pH dependence of their interactions with palmitoyloleoyl phosphatidylcholine (POPC) and palmitoyloleoyl phosphatidylglycerol (POPG) membranes was studied. A filtration binding assay shows that acidic partly folded states of BCAII bind to POPC membranes. Fluorescence emission spectra from Trp residues of the bound protein are slightly shifted to shorter wavelength and can be quenched by a water-soluble quencher of fluorescence, indicating that the binding occurs without deep penetration of Trp residues into the membrane. The content of beta-structures of the protein in solution, as revealed by FT-IR spectroscopy, decreases in the partly folded states and the binding to POPC membrane occurs without further changes of secondary structure. In the presence of 0.1 M NaCl, a partly folded state self aggregates and does not bind to POPC membrane. At acidic pH, BCAII binds to POPG membranes both at high and low ionic strength. The binding to the anionic lipid occurs with protein self-aggregation within the lipid-protein complexes and with changes in the secondary structure; large blue shifts in the fluorescence emission spectra and the decrease in the exposure to water-soluble acrylamide quencher of Trp fluorescence strongly suggest that BCAII penetrates the hydrocarbon domain in the POPG-protein complexes. PMID- 11018658 TI - Stabilization of lipid/DNA complexes during the freezing step of the lyophilization process: the particle isolation hypothesis. AB - The instability of nonviral vectors in aqueous suspensions has stimulated an interest in developing lyophilized formulations for use in gene therapy. Previous work has demonstrated a strong correlation between the maintenance of particle size and retention of transfection rates. Our earlier work has shown that aggregation of nonviral vectors typically occurs during the freezing step of the lyophilization process, and that high concentrations of sugars are capable of maintaining particle size. This study extends these observations, and demonstrates that glass formation is not the mechanism by which sugars protect lipid/DNA complexes during freezing. We also show that polymers (e.g., hydroxyethyl starch) are not capable of preventing aggregation despite their ability to form glasses at relatively high subzero temperatures. Instead, our data suggest that it is the separation of individual particles within the unfrozen fraction that prevents aggregation during freezing, i.e., the particle isolation hypothesis. Furthermore, we suggest that the relatively low surface tension of mono- and disaccharides, as compared to starch, allows phase-separated particles to remain dispersed within the unfrozen excipient solution, which preserves particle size and transfection rates during freezing. PMID- 11018660 TI - Analysis of chlorophyll-protein complexes from the cyanobacterium Cyanothece sp. ATCC 51142 by non-denaturing gel electrophoresis. AB - The unicellular diazotrophic cyanobacterium, Cyanothece sp. ATCC 51142 temporally separates N(2) fixation from photosynthesis. We are analyzing the mechanism by which photosynthesis is down-regulated so that O(2) evolution is minimized during N(2) fixation. Previous results suggested changes in photosynthesis that are mediated through the redox poise of the plastoquinone pool (a process involving state transitions, in which the redistribution of excitation energy between the two photosystems helps to optimize photosynthetic yield) and the oligomerization state of the photosystems. Our working hypothesis was that the regulation of photosynthesis involved changes in the oligomerization of the photosystems. To analyze this hypothesis, we utilized a low-ionic strength, non-denaturing gel electrophoresis system to study the Chl-protein complexes. We determined that PSI is mostly trimeric, whereas PSII appears mainly as monomers. We demonstrated that most of the Chl-protein complexes in Cyanothece sp. remained constant throughout the diurnal cycle, except for the transient accumulation of a Chl-protein complex (band C) which appeared only during the late light period. Based on the size of this complex, band C represents either an interaction of PSI and PSII or a PSII dimer. These results provide support for the dynamic nature of the photosystems with respect to the diurnal cycle. PMID- 11018659 TI - The quantitative analysis of three action modes of volatile anesthetics on purple membrane. AB - We quantitatively assessed the spectroscopic changes of purple membrane in relation to the concentrations of a volatile anesthetic. As reported previously, volatile anesthetics show three modes of action on purple membrane. By using an anesthetic for which the concentration in solution could be determined spectroscopically and by applying modified analytical methods regarding the M intermediate lifetime, we were able to clarify the quantitative relation between anesthetic concentration and each mode of action, a relation which in the past has only been described qualitatively. We also determined through the measurement of transient pH changes with pyranine that the proton pump efficiency per photochemical cycle in an action mode induced with low concentrations of anesthetic does not change from that of the native state. Moreover, we dynamically obtained the individual M-bacteriorhodopsin difference spectrum of each state at room temperature using our flash photolysis system equipped with a wavelength-tunable dye laser. These results demonstrated again that we should clearly distinguish different action modes of anesthetics according to their concentrations. PMID- 11018662 TI - Physical properties of liposomes and proteoliposomes prepared from Escherichia coli polar lipids. AB - Reconstituted proteoliposomes serve as experimental systems for the study of membrane enzymes. Osmotic shifts and other changes in the solution environment may influence the structures and membrane properties of phospholipid vesicles (including liposomes, proteoliposomes and biological membrane vesicles) and hence the activities of membrane-associated proteins. Polar lipid extracts from Escherichia coli are commonly used in membrane protein reconstitution. The solution environment influenced the phase transition temperature and the diameter of liposomes and proteoliposomes prepared from E. coli polar lipid by extrusion. Liposomes prepared from E. coli polar lipids differed from dioleoylphosphatidylglycerol liposomes in Young's elastic modulus, yield point for solute leakage and structural response to osmotic shifts, the latter indicated by static light scattering spectroscopy. At high concentrations, NaCl caused aggregation of E. coli lipid liposomes that precluded detailed interpretation of light scattering data. Proteoliposomes and liposomes prepared from E. coli polar lipids were similar in size, yield point for solute leakage and structural response to osmotic shifts imposed with sucrose as osmolyte. These results will facilitate studies of bacterial enzymes implicated in osmosensing and of other enzymes that are reconstituted in E. coli lipid vesicles. PMID- 11018661 TI - Sterically stabilized liposomes bearing anti-HLA-DR antibodies for targeting the primary cellular reservoirs of HIV-1. AB - The ability of liposomes bearing anti-HLA-DR Fab' fragments at the end termini of polyethyleneglycol chains (sterically stabilized immunoliposomes) to target HLA DR expressing cells and increase the accumulation of liposomes into lymphoid organs has been evaluated and compared to that of conventional liposomes, sterically stabilized liposomes and conventional immunoliposomes after a single subcutaneous injection to mice. The accumulation of sterically stabilized liposomes in lymph nodes was higher than that of conventional liposomes. Sterically stabilized immunoliposomes accumulated much better than conventional immunoliposomes in all tissues indicating that the presence of PEG has an important effect on the uptake of immunoliposomes by the lymphatic system. Fluorescence microscopy studies showed that sterically stabilized liposomes are mainly localized in macrophage-rich areas such as the subcapsular region of lymph nodes and in the red pulp and marginal zone of the spleen. In contrast, sterically stabilized immunoliposomes mostly accumulated in the cortex in which follicles are located and in the white pulp of the spleen. As the human HLA-DR determinant of the major histocompatibility complex class II is expressed on activated CD4+ T lymphocytes and antigen presenting cells such as monocyte/macrophages and dendritic cells, known as the cellular reservoirs of HIV 1, liposomes bearing anti-HLA-DR antibodies constitute an attractive approach to concentrate drugs in HIV-1 reservoirs and improve their therapeutic effect. PMID- 11018663 TI - A solid-state NMR study of the phospholamban transmembrane domain: local structure and interactions with Ca(2+)-ATPase. AB - The structure and dynamics of a double (13)C-labelled 24-residue synthetic peptide ([(13)C(2)]CAPLB(29-52)), corresponding to the membrane-spanning sequence of phospholamban (PLB), were examined using (13)C cross-polarisation magic-angle spinning (CP-MAS) NMR spectroscopy. CP-MAS spectra of [(13)C(2)]CAPLB(29-52) reconstituted into unsaturated lipid membranes indicated that the peptide was mobile at temperatures down to -50 degrees C. The NMR spectra showed that peptide motion became constrained in the presence of the SERCA1 isoform of Ca(2+)-ATPase, and chemical cross-linking experiments indicated that [(13)C(2)]CAPLB(29-52) and Ca(2+)-ATPase came into close contact with one another. These results together suggested that the peptide and the 110-kDa calcium pump were interacting in the membrane. Rotational resonance CP-MAS (13)C-(13)C distance measurements on [(13)C(2)]CAPLB(29-52) reconstituted into lipid bilayers confirmed that the sequence spanning Phe-32 and Ala-36 was alpha-helical, and that this structure was not disrupted by interaction with Ca(2+)-ATPase. These results support the finding that the transmembrane domain of PLB is partially responsible for regulation of Ca(2+) transport through interactions with cardiac muscle Ca(2+) ATPase in the lipid bilayer, and also demonstrate the feasibility of performing structural measurements on PLB peptides when bound to their physiological target. PMID- 11018664 TI - Organization of water molecules by adhering to oriented layers of dipalmitoylphosphatidyl serine in the presence of varying concentrations of cholesterol. AB - About seven water molecules adhere to one molecule of dipalmitoylphosphatidyl serine (DPPS) in an oriented surface layer as inferred from the increase of the dichroic ratio R of their OH stretching vibration band (3400 cm(-1)) from 2 in the random bulk state to about 2.8 when adhering to DPPS. In DPPS-cholesterol mixtures the number of water molecules adhering to the phospholipid molecules and oriented by them increases as cholesterol content increases. This increase is very steep between molar fractions of cholesterol X(chol)=0.2-0.4 and at X(chol)=0.6 about 13 water molecules adhere and are oriented by one DPPS molecule. PMID- 11018665 TI - Fast K(+) currents from cerebellum granular cells are completely blocked by a peptide purified from Androctonus australis Garzoni scorpion venom. AB - A novel peptide was purified from the venom of the scorpion Androctonus australis Garzoni (abbreviated Aa1, corresponding to the systematic number alpha KTX4.4). It contains 37 amino acid residues, has a molecular mass of 3850 Da, is closely packed by three disulfide bridges and a blocked N-terminal amino acid. This peptide selectively affects the K(+) currents recorded from cerebellum granular cells. Only the fast activating and inactivating current, with a kinetics similar to I(A)-type current, is completely blocked by the addition of low micromolar concentrations (K(i) value of 150 nM) of peptide Aa1 to the external side of the cell preparation. The blockade is partially reversible in our experimental conditions. Aa1 blocks the channels in both the open and the closed states. The blockage is test potential independent and is not affected by changes in the holding potential. The kinetics of the current are not affected by the addition of Aa1 to the preparation; it means that the block is a simple 'plugging mechanism', in which a single toxin molecule finds a specific receptor site in the external vestibule of the K(+) channel and thereby occludes the outer entry to the K(+) conducting pore. PMID- 11018666 TI - X-ray studies on the interaction of the antimicrobial peptide gramicidin S with microbial lipid extracts: evidence for cubic phase formation. AB - We have investigated the effect of the interaction of the antimicrobial peptide gramicidin S (GS) on the thermotropic phase behavior of model lipid bilayer membranes generated from the total membrane lipids of Acholeplasma laidlawii B and Escherichia coli. The A. laidlawii B membrane lipids consist primarily of neutral glycolipids and anionic phospholipids, while the E. coli inner membrane lipids consist exclusively of zwitterionic and anionic phospholipids. We show that the addition of GS at a lipid-to-peptide molar ratio of 25 strongly promotes the formation of bicontinuous inverted cubic phases in both of these lipid model membranes, predominantly of space group Pn3m. In addition, the presence of GS causes a thinning of the liquid-crystalline bilayer and a reduction in the lattice spacing of the inverted cubic phase which can form in the GS-free membrane lipid extracts at sufficiently high temperatures. This latter finding implies that GS potentiates the formation of an inverted cubic phase by increasing the negative curvature stress in the host lipid bilayer. This effect may be an important aspect of the permeabilization and eventual disruption of the lipid bilayer phase of biological membranes, which appears to be the mechanism by which GS kills bacterial cells and lysis erythrocytes. PMID- 11018667 TI - Endogenous cleavage of annexin I generates a truncated protein with a reduced calcium requirement for binding to neutrophil secretory vesicles and plasma membrane. AB - We have earlier shown that an N-terminal truncated annexin I molecule, annexin I(des1-8), is generated in human neutrophils through cleavage by a membrane localized metalloprotease. The truncated protein showed differences in membrane binding among the neutrophil granule populations as compared to full-length annexin I. In this study, we investigated the cleavage capabilities of isolated neutrophil secretory vesicles and plasma membrane, and the binding of full-length annexin I and annexin I(des1-8) to these membrane fractions. Translocations were performed in vitro to secretory vesicles and plasma membrane, respectively, at different Ca(2+) concentrations. We show that the annexin I-cleaving membrane localized metalloprotease is present both in the secretory vesicles and the plasma membrane. The N-terminal truncation of annexin I gives rise to a molecule with a decreased Ca(2+) requirement for binding, both to secretory vesicles and plasma membrane. There was, thus, no difference in binding of either full-length annexin I or annexin I(des1-8) to the secretory vesicles as compared to the plasma membrane. PMID- 11018668 TI - Efficient encapsulation of DNA plasmids in small neutral liposomes induced by ethanol and calcium. AB - Efficient encapsulation of DNA plasmids inside small, neutral liposomes composed of 1,2-dioleoyl-sn-phosphatidylcholine (DOPC), DOPC/DOPE (1,2-dioleoyl-sn phosphatidylethanolamine) (1:1) and DOPC/DOPE/cholesterol (1:1:1) was achieved by the addition of ethanol and calcium chloride to an aqueous mixture of small unilamellar vesicles (SUVs) and plasmid. Following dialysis against low-salt buffer, the neutral lipid complexes (NLCs) had average effective diameters less than 200 nm and encapsulated up to 80% of the DNA. Optimum Ca(2+) and ethanol concentrations for each lipid mixture were determined by statistically designed experiments and mathematical modeling of trapping efficiency. NLCs are unilamellar, have neutral surface potentials, and retain entrapped DNA at pH 4.0 and in serum at 37 degrees C. The circulation and clearance properties of the complexes following intravenous administration in mice are similar to empty neutral liposomes, and the toxicity of NLCs are expected to be significantly reduced compared to other non-viral gene-delivery systems. The NLC encapsulation method, if it can be combined with effective targeting and endosome-release technologies to achieve efficient and tissue-specific transfection, may represent an important alternative to current systemic gene therapy approaches. PMID- 11018669 TI - Localization of sterically stabilized liposomes in experimental rat Klebsiella pneumoniae pneumonia: dependence on circulation kinetics and presence of poly(ethylene)glycol coating. AB - Preferential localization of liposomes at sites of infection or inflammation has been demonstrated in a variety of experimental models. Most studies report enhanced localization at the target site of poly(ethyelene) glycol (PEG)-coated liposomes as compared to conventional non-coated liposomes. It is generally accepted that the prolonged circulation time of PEG-coated liposomes increases target site exposure, which results in increased target localization. A quantitative relationship between circulation kinetics and localization at the pathological site has not been defined as yet. Besides, an effect of the PEG coating itself has been suggested, as theoretically the PEG coating may facilitate liposome extravasation. In the present study, in a rat model of an acute unilateral Klebsiella pneumoniae pneumonia, circulation kinetics of PEG coated liposomes were manipulated by incorporation of different amounts of phosphatidylserine (PS) and variation of lipid dose, additionally allowing evaluation of the saturability of the localization process. In addition, this paper addresses the effect of the PEG coating, by comparing the circulation kinetics and target localization of long-circulating 'PEG-free' and PEG-coated liposomes. It is shown that the degree of liposome localization at the target site is positively linearly related to the area under the blood concentration time curve (AUC) of the liposome formulations, irrespective of PEG coating. This finding is discussed in relation to the equation of Kedem and Katchalsky, which describes protein influx into sites of infection or inflammation. PMID- 11018670 TI - A transmembrane peptide from the human EGF receptor: behaviour of the cytoplasmic juxtamembrane domain in lipid bilayers. AB - Solid state (2)H NMR spectroscopy was employed to study peptides related to the transmembrane domain of the human epidermal growth factor receptor, for insight into the interaction of its cytoplasmic juxtamembrane domain with the membrane surface. Since such receptors have clusters of (+)charged amino acids in this region, the effect of (-)charged phosphatidylserine at the concentration found naturally in the cytoplasmic leaflet (15 mol%) was considered. Each peptide contained 34 amino acids, which included the hydrophobic 23 amino acid stretch thought to span the membrane and a ten amino acid segment beyond the 'cytoplasmic' surface. Non-perturbing deuterium probe nuclei were located within alanine side chains in intramembranous and extramembranous portions. (2)H NMR spectra were recorded at 35 degrees C and 65 degrees C in fluid lipid bilayers consisting of (zwitterionic) 1-palmitoyl-2-oleoylphosphatidylcholine, with and without 15 mol% (anionic) phosphatidylserine. The cationic extramembranous portion of the receptor backbone was found to be highly rotationally mobile on a time scale of 10(-4)-10(-5) s in both types of membrane - as was the alpha helical intramembranous portion. Deuterium nuclei in alanine side chains (-CD(3)) detected modest changes in peptide backbone orientation and/or dynamics related to the presence of 1-stearoyl-2-oleoylphosphatidylserine: in the case of the extramembranous portion of the peptide these seemed related to lipid charge. Temperature effects on the peptide backbone external to the membrane were qualitatively different from effects on the helical transmembrane domain - likely reflecting the different physical constraints on these peptide regions and the greater flexibility of the extramembranous domain. Effects related to lipid charge could be detected in the spectrum of CD(3) groups on the internally mobile side chain of Val(650), six residues beyond the membrane surface. PMID- 11018671 TI - Use of sulfhydryl reagents to investigate branched chain alpha-keto acid transport in mitochondria. AB - The goal of this paper was to determine the contribution of the mitochondrial branched chain aminotransferase (BCATm) to branched chain alpha-keto acid transport within rat heart mitochondria. Isolated heart mitochondria were treated with sulfhydryl reagents of varying permeability, and the data suggest that essential cysteine residues in BCATm are accessible from the cytosolic face of the inner membrane. Treatment with 15 nmol/mg N-ethylmaleimide (NEM) inhibited initial rates of alpha-ketoisocaproate (KIC) uptake in reconstituted mitochondrial detergent extracts by 70% and in the intact organelle by 50%. KIC protected against inhibition suggesting that NEM labeled a cysteine residue that is inaccessible when substrate is bound to the enzyme. Additionally, the apparent mitochondrial equilibrium KIC concentration was decreased 50-60% after NEM labeling, and this difference could not be attributed to effects of NEM on matrix pH or KIC oxidation. In fact, NEM was a better inhibitor of KIC oxidation than rotenone. Measuring matrix aspartate and glutamate levels revealed that the effects of NEM on the steady-state KIC concentration resulted from inhibition of BCATm catalyzed transamination of KIC with matrix glutamate to form leucine. Furthermore, circular dichroism spectra of recombinant human BCATm with liposomes showed that the commercial lipids used in the reconstituted transport assay contain BCAT amino acid substrates. Thus BCATm is distinct from the branched chain alpha-keto acid carrier but may interact with the inner mitochondrial membrane, and it is necessary to inhibit or remove transaminase activity in both intact and reconstituted systems prior to quantifying transport of alpha-keto acids which are transaminase substrates. PMID- 11018672 TI - Anion antiport mechanism is involved in transport of lactic acid across intestinal epithelial brush-border membrane. AB - Intestinal epithelial membrane transport of L-lactic acid was characterized using rabbit jejunal brush-border membrane vesicles (BBMVs). The uptake of L [(14)C]lactic acid by BBMVs showed an overshoot phenomenon in the presence of outward-directed bicarbonate and/or inward-directed proton gradients. Kinetic analysis of L-[(14)C]lactic acid uptake revealed the involvement of two saturable processes in the presence of both proton and bicarbonate gradients. An arginyl residue-modifying agent, phenylglyoxal, inhibited L-[(14)C]lactic acid transport by the proton cotransporter, but not by the anion antiporter. The initial uptakes of L-[(14)C]lactic acid which are driven by bicarbonate ion and proton gradients were inhibited commonly by monocarboxylic acids and selectively by anion exchange inhibitor 4, 4'-diisothiocyanostilbene-2,2'-disulfonic acid and protonophore carbonylcyanide p-trifluoromethoxyphenylhydrazone, respectively. These observations demonstrate that L-lactic acid is transported across the intestinal brush-border membrane by multiple mechanisms, including an anion antiporter and a previously known proton cotransporter. PMID- 11018674 TI - Effect of fenitrothion on dipalmitoyl and 1-palmitoyl-2-oleoylphosphatidylcholine bilayers. AB - The effects of the organophosphorous insecticide fenitrothion (phosphorothioic acid, O,O-dimethyl O-(3-methyl-4-nitrophenyl) ester; FS) on the physical state of pure dipalmitoyl (DPPC) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) membranes were investigated. FS lowers the phase transition temperature of DPPC. It has no large effects on the DPPC gel phase, but it increases the order of the liquid-crystalline state of DPPC and POPC. FS also decreases 1,6-diphenyl-1,3,5 hexatriene (DPH) lifetime (tau) in the DPPC and POPC liquid-crystalline states. Since a direct quenching of DPH emission by FS was ruled out, tau shortening is assigned to an increased water penetration in the bilayer. The effect of FS is different from most perturbing agents for which an increased order is accompanied by a higher tau. Furthermore, quenching of DPH by KI was increased by FS in POPC liposomes indicating an increased accessibility of the quencher to the hydrophobic core where DPH distributes. The effect of FS on dipole relaxation at the hydrophilic-hydrophobic interface of POPC bilayers was studied with 2 dimethylamino-6-lauroylnaphthalene (Laurdan). FS produces a decrease in Laurdan tau and a narrowing of its emission band. FS significantly increases the generalized polarization values at both emission band ends. These results indicate that FS may allow the coexistence of microdomains that have different physical properties. PMID- 11018673 TI - Fourier transform infrared spectroscopy and differential scanning calorimetry studies of fatty acid homogeneous ceramide 2. AB - Ceramides provide a major component of the barrier function of skin. An understanding of barrier organization requires a detailed characterization of ceramide phase behavior and molecular interactions. Toward this end, Fourier transform infrared (FTIR) and differential scanning calorimetry (DSC) studies of ceramide 2 analogues (non-hydroxylated fatty acid N-acyl sphingosines) of specific chain lengths (C(14), C(16), C(18), C(20)) are presented. In addition, the molecular interactions of the individual chains in each molecule are elucidated through thermotropic FTIR studies of derivatives possessing perdeuterated fatty acid chains. DSC data showed a much smaller chain length variation (for the C(16), C(18), C(20) derivatives) in the main order-disorder transition temperature (approx. 93+/-1 degrees C) than is observed in the corresponding series of phosphatidylcholines, consistent with minimal ceramide hydration. The temperature dependence of the methylene stretching and scissoring modes revealed a solid-solid phase transition at 20-25 degrees C below the main order-disorder transition accompanied by chain packing alterations from orthorhombic-->hexagonal subcells. The chain packing transition was accompanied by enhanced penetration of water into the polar region. This was deduced from the temperature dependence of the amide I and II modes, which provide direct evidence for H-->D exchange. The CD(2) scissoring mode splitting of the deuterated fatty acid constituent of the C(16), C(18), C(20) chains revealed preferential segregation of microdomains (3-5 chains) of this species within the orthorhombic phase. In contrast, the sphingosine base chains appeared to be sufficiently separated so as to inhibit interchain vibrational coupling between them. FTIR spectroscopy provides a convenient means for characterizing domain formation, chain packing, and hydration sites of these phases, which are highly ordered under physiological conditions. PMID- 11018675 TI - Glucose-sensitive conductances in rat pancreatic beta-cells: contribution to electrical activity. AB - The perforated patch technique was used to assess the relative contribution of K(ATP) channel activity, assessed from input conductance (G(input)), and volume sensitive anion channel activity to the induction of electrical activity in single isolated rat pancreatic beta-cells by glucose, 2-ketoisocaproate and tolbutamide. In cells equilibrated in the absence of glucose, the membrane potential was -71 mV and G(input) 3.66 nS. Addition of 8 mM glucose resulted in depolarisation, electrical activity and a reduction in G(input), reflecting an inhibition of K(ATP) channels. Cells equilibrated in 4 mM glucose had a membrane potential of -59 mV and a G(input) of 0.88 nS. In this case, a rise in glucose concentration to 8-20 mM again resulted in depolarisation and electrical activity, but caused a small increase in G(input). 2-Ketoisocaproate also evoked electrical activity and an increase in G(input), whereas electrical activity elicited by addition of tolbutamide was accompanied by reduced G(input). Increasing the concentration of glucose from 4 to 8-20 mM generated a noisy inward current at -70 mV, reflecting activation of the volume-sensitive anion channel. The mean amplitude of this current was glucose-dependent within the range 4-20 mM. Addition of 2-ketoisocaproate or a 15% hypotonic solution elicited similar increases in inward current. In contrast, addition of tolbutamide failed to induce the inward current. It is concluded that K(ATP) channel activity is most sensitive to glucose within the range 0-4 mM. At higher glucose concentrations effective in generating electrical activity, activation of the volume-sensitive anion channel could contribute towards the nutrient-induced increase in G(input). PMID- 11018676 TI - Protonation-dependent inactivation of Na,K-ATPase by hydrostatic pressure developed at high-speed centrifugation. AB - Irreversible inactivation of membranous Na,K-ATPase by high-speed centrifugation in dilute aqueous solutions depends markedly on the protonation state of the protein. Pig kidney Na,K-ATPase is irreversibly inactivated at pH 5 but is fully protected at pH 7 and above. Shark rectal gland Na,K-ATPase is irreversibly inactivated at neutral or acidic pH and partially protected at an alkaline pH. The overall Na,K-ATPase activity and the K-dependent pNPPase activity were denatured in parallel. Cryoprotectants such as glycerol or sucrose at concentrations of 25-30% fully protect both enzymes against inactivation. The specific ligands NaCl and KCl protect the Na,K-ATPase activity partially and the pNPPase activity fully at concentrations of 0.2-0.3 M. Electron microscope analysis of the centrifuged Na,K-ATPase membranes revealed that the ultrastructure of the native membranes is preserved upon inactivation. It was also observed that the sarcoplasmic reticulum Ca-ATPase and hog gastric H, K ATPase are susceptible to inactivation by high-speed centrifugation in a pH dependent fashion. H,K-ATPase is protected at alkaline pH, whereas Ca-ATPase is protected only in the neutral pH range. PMID- 11018677 TI - alpha-methylene ordering of acyl chains differs in glucolipids and phosphatidylglycerol from Acholeplasma laidlawii membranes: (2)H-NMR quadrupole splittings from individual lipids in mixed bilayers. AB - A Acholeplasma laidlawii strain A-EF22 was grown in a medium supplemented with alpha-deuterated oleic acid. Phosphatidylglycerol (PG), the glucolipids monoglucosyldiacylglycerol (MGlcDAG), diglucosyldiacylglycerol (DGlcDAG) and monoacyldiglucosyldiacylglycerol, and the phosphoglucolipid glycerophosphoryldiglucosyldiacylglycerol (GPDGlcDAG) were purified, and the phase behaviour and molecular ordering for the individual lipids, as well as for mixtures of the lipids, were studied by (2)H-, (31)P-NMR and X-ray scattering methods. The chemical structure of all the A. laidlawii lipids, except PG, has been determined and verified previously; here also the chemical structure of PG was verified, utilising mass spectrometry and (1)H and (13)C high resolution NMR spectroscopy. For the first time, lipid dimers were found in the mass spectrometry measurements. The major findings in this work are: (1) addition of 50 mol% of PG to the non-lamellar-forming lipid MGlcDAG does not significantly alter the transition temperature between lamellar and non-lamellar phases; (2) the (2)H-NMR quadrupole splitting patterns obtained from the lamellar liquid crystalline phase are markedly different for PG on one hand, and DGlcDAG and GPDGlcDAG on the other hand; and (3) mixtures of PG and DGlcDAG or MGlcDAG give rise to (2)H-NMR spectra consisting of a superposition of splitting patterns of the individual lipids. These remarkable features show that the local ordering of the alpha-carbon of the acyl chains is different for PG than for MGlcDAG and DGlcDAG, and that this difference is preserved when PG is mixed with the glucolipids. The results obtained are interpreted in terms of differences in molecular shape and hydrophilicity of the different polar headgroups. PMID- 11018679 TI - Comparative dynamics and location of chain spin-labelled sphingomyelin and phosphatidylcholine in dimyristoyl phosphatidylcholine membranes studied by EPR spectroscopy. AB - The dynamics and environment of sphingomyelin spin-labelled at different positions in the N-acyl chain have been studied in dimyristoyl phosphatidylcholine bilayer membranes by using electron spin resonance spectroscopy. Comparison was made with phosphatidylcholine spin-labelled on the sn-2 acyl chain in the same host membrane. Spin-labelled sphingomyelin was found to mix well with the host phosphatidylcholine lipids in both gel and fluid phase membranes. At 1 mol%, mutual spin-spin interactions are no greater than for spin labelled phosphatidylcholine. In the fluid membrane phase, the effective chain order parameters and polarity-sensitive isotropic hyperfine coupling constants of spin-labelled sphingomyelin display a similar dependence on the position of labelling to those of spin-labelled phosphatidylcholine. The values of both parameters are, however, generally larger for sphingomyelin than for phosphatidylcholine at equivalent positions of acyl chain labelling. This difference is attributed to the different chain linkage of sphingo- and glycero lipids, combined with an offset of approximately one C-atom in transbilayer register between the respective N-acyl and O-acyl chains. In the gel phase, differences in chain configuration between sphingomyelin and phosphatidylcholine are indicated by differences in spin label spectral anisotropy between the two lipids, which appears to reverse towards the terminal methyl chain end. PMID- 11018678 TI - The effect of Zn(2+) on the secondary structure of a histidine-rich fusogenic peptide and its interaction with lipid membranes. AB - Membrane fusion between uncharged lipid vesicles can be triggered by the peptide sequence 'B18' from the fertilization protein 'bindin', but it only proceeds efficiently in the presence of Zn(2+) ions. We studied (i) the interaction of Zn(2+) with the fusogenic peptide B18, (ii) the binding of B18 to 1-palmitoyl-2 oleoylglycero-3-phosphocholine (POPC), and (iii) the ternary system POPC/B18/Zn(2+). The complex formation of Zn(2+) with the central histidine-rich motif of B18 appears to shift the secondary structure away from a beta-sheet towards an alpha-helical conformation. Here we observe for the first time an essentially alpha-helical structure of the peptide when immersed in POPC bilayers which appears to represent its functional fusogenic state. Infrared linear dichroism suggests a peripheral, oblique insertion mode of B18, mediated by the hydrophobic patches along one side of the amphipathic peptide. Furthermore, the hydration level of the peptide is reduced, suggesting that the hydrophobic region of the bilayer is involved in the lipid/peptide interactions. The hydration capacity of the POPC/B18/Zn(2+) system is distinctly smaller than that of POPC/Zn(2+) without peptide. The accompanying decrease in the number of tightly bound water molecules per lipid can be interpreted as a reduction in the repulsive 'hydration' forces, which usually prevent the spontaneous fusion of lipid vesicles. Binding of the B18 peptide in the presence of Zn(2+) effectively renders the membrane surface more hydrophobic, thus allowing fusion to proceed. PMID- 11018680 TI - The transport modifier RS1 is localized at the inner side of the plasma membrane and changes membrane capacitance. AB - Previously we cloned membrane associated (M(r) 62000-67000) polypeptides from pig (pRS1), rabbit (rbRS1) and man (hRS1) which modified transport activities that were expressed in Xenopus laevis oocytes by the Na(+)-D-glucose cotransporter SGLT1 and/or the organic cation transporter OCT2. These effects were dependent on the species of RS1 and on the target transporters. hRS1 and rbRS1 were shown to be intronless single copy genes which are expressed in various tissues and cell types. Earlier immunohistochemical data with a monoclonal IgM antibody suggested an extracellular membrane association of RS1. In the present paper antibodies against recombinant pRS1 were raised and the distribution and membrane localization of RS1 reevaluated. After subcellular fractionation of renal cortex RS1 was found associated with brush border membranes and an about 1:200 relation between RS1 and SGLT1 protein was estimated. Also after overexpression in X. laevis oocytes RS1 was associated with the plasma membrane, however, at variance to the kidney it was also observed in the cytosol. Labeling experiments with covalently binding lipid-permeable and lipid-impermeable biotin analogues showed that RS1 is localized at the inner side of the plasma membrane. Western blots with plasma membranes from Xenopus oocytes revealed that SGLT1 protein in the plasma membrane was reduced when hRS1 was coexpressed with human SGLT1 which leads to a reduction in V(max) of expressed glucose transport. Measurements of membrane capacitance and electron microscopic inspection showed that the expression of hRS1 leads to a reduction of the oocyte plasma membrane surface. The data suggest that RS1 is an intracellular regulatory protein that associates with the plasma membrane. Overexpression of RS1 may effect the incorporation and/or retrieval of transporters into the plasma membrane. PMID- 11018681 TI - The interaction between lipid derivatives of colchicine and tubulin: consequences of the interaction of the alkaloid with lipid membranes. AB - Colchicine is a potent antimitotic poison which is well known to prevent microtubule assembly by binding tubulin very tightly. Colchicine also possesses anti-inflammatory properties which are not well understood yet. Here we show that colchicine tightly interacts with lipid layers. The physical and biological properties of three different lipid derivatives of colchicine are investigated parallel to those of membrane lipids in the presence of colchicine. Upon insertion in the fatty alkyl chains, colchicine rigidifies the lipid monolayers in a fluid phase and fluidifies rigid monolayers. Similarly X-ray diffraction data show that lecithin-water phases are destabilized by colchicine. In addition, an unexpectedly drastic enhancement of the photoisomerization rate of colchicine into lumicolchicine in the lipid environment is observed and further supports insertion of the alkaloid in membranes. Finally the interaction of colchicine with lipids makes the drug inaccessible to tubulin. The possible in vivo significance of these results is discussed. PMID- 11018682 TI - Gene transfection activities of amphiphilic steroid-polyamine conjugates. AB - The design and evaluation of a novel potent class of DNA delivery agents based on steroid-polyamine conjugates bearing a flexible linker are reported. The hydrophobic regions are based on steroids, i.e. chlolestane and lithocholic acid motifs. The linker, which couples a hydrophobic steroid and a hydrophilic polyamine, in this study can be regarded as a two-atom extension of the conventional carbamate linker. We found that the gene transfection activity of the steroid-polyamine conjugates is influenced by the polyamine chain length and steroid structure. Molecular modeling of the relevant amphiphilic molecules revealed low-energy structures in which the polyamine chains are folded rather than stretched. This work suggests a significant effect of space-filling, i.e. the shape and orientation of the hydrophilic and hydrophobic regions, upon the efficiency of gene transfection. PMID- 11018683 TI - Mini-review: therapeutic drug monitoring in pediatrics. PMID- 11018684 TI - An Ontario-wide study of vitamin B12, serum folate, and red cell folate levels in relation to plasma homocysteine: is a preventable public health issue on the rise?. AB - BACKGROUND: Plasma homocysteine has been reported to be useful in the evaluation of patients with suspected vitamin B12 or folate deficiency. In November 1998, Canada began its mandatory fortification of all flour, and some corn and rice products, with folic acid. We evaluated the status of folate and vitamin B12 in Ontario since this fortification program began, and also studied the role of plasma homocysteine in the assessment of vitamin B12 deficiency since that time. METHODS: A retrospective cross-sectional study design was performed using a community database of all Ontario samples analyzed by MDS Laboratories, a major provider of diagnostic laboratory services in Canada. All consecutive single patient fasting samples for plasma homocysteine collected between January 1 and September 30, 1999 were included, as well as corresponding red cell folate and serum B12 concentrations. Data for serum folate were included when available. Descriptive statistics included the arithmetic and geometric means for each measure, as well as the lower and upper centile values. After excluding cases with a concomitant serum creatinine > 120 micromol/L or red cell folate < 215 nmol/L, we established the test properties of a plasma homocyteine level of 15 micromol/L or greater for the diagnosis of "low" (< 120 pmol/L) or "indeterminate" (i.e., between 120 and 150 pmol/L) serum vitamin B12 concentrations. RESULTS: The mean age of all subjects was 58.4 years (95% CI 57.4 to 59.4). Plasma homocysteine samples were obtained from 403 males (56.7%) and 308 females. The geometric mean homocysteine concentration for the entire population was 8.3 micromol/L, and was significantly higher among males (9.3 micromol/L) than females (8.3 micromol/L) (unpaired t-test: 2-p < 0.0001). The geometric mean serum folate concentration was significantly higher in females (35.8 nmol/L) than males (33.6 nmol/L) (2-p < 0.0001), as were the mean red cell folate levels (females 966.8 nmol/L, males 949.3 nmol/L; 2-p < 0.0001). Serum vitamin B12 concentrations were available for 692 subjects, with a geometric mean of 322.0 pmol/L. Again, mean vitamin B12 was higher in females (332.5 pmol/L) than males (314.3 pmol/L) (2-p < 0.0001). The fifth centile for vitamin B12 was 134.6 pmol/L. A plasma homocysteine concentration > 15 micromol/L did not discriminate between cobalamin concentrations below versus above 120 pmol/L (positive and negative predictive values 7.4% and 97.2%, respectively), nor did it discriminate "indeterminate" B12 levels between 120 and 150 pmol/L (positive and negative predictive values 6.3% and 94.0%, respectively). CONCLUSION: In a large select group of Ontarians, serum and red cell folate concentrations appear to be higher than expected, possibly due to a recent national folate fortification programme; cobalamin levels are no higher than expected. Given our inability to detect mild B12 deficiency using such indicators as plasma homocysteine, and considering the substantial growth in the elderly segment of the Canadian population, occult cobalamin deficiency could become a common disorder. Accordingly, we recommend either consideration of the addition of vitamin B12 to the current folate fortification programme, and/or the development of better methods for the detection of cobalamin deficiency. PMID- 11018685 TI - Polyvinylamine-streptavidin complexes labeled with a europium chelator: a universal detection reagent for solid-phase time resolved fluorometric applications. AB - OBJECTIVES: To describe the synthesis and characteristics of a new streptavidin based universal detection reagent which is multiply labeled with the europium chelate of 4,7-bis(chlorosulfophenyl)-1, 10-phenanthroline-2,9-dicarboxylic-acid (BCPDA). METHODS AND RESULTS: Polyvinylamine (PVA) was first labeled with biotin(b) and then with BCPDA to create (b)(x)-PVA-(BCPDA)(y). By mixing controlled amounts of this complex with streptavidin (SA) and a fixed amount of Eu(3+), we were able to produce the conjugate (SA)(z)-(b)(x)-PVA(BCPDA)(y) Eu(3+). This conjugate is reactive, highly fluorescent, and stable for at least 12 months. It was used to develop model solid-phase time-resolved fluoroimmunoassays for biotinylated mouse IgG and prostate specific antigen (PSA). Detection limits achieved were around 1 to 2 ng/L ( approximately 3 x 10( 18) moles/assay). CONCLUSIONS: A new universal detection reagent was synthesized, which can be used in combination with biotinylated reagents (e.g., antibodies, DNA probes, etc.) for the development of highly sensitive solid-phase time resolved fluorescence-based assays. PMID- 11018686 TI - Effect of CDP-choline on brain acetylcholinesterase and Na(+), K(+)-ATPase in adult rats. AB - OBJECTIVE: The aim of this work was to investigate the effect of different cytidine-5'-diphosphocholine (CDP-choline) concentrations (0.1-1 mM) on acetylcholinesterase (AChE), (Na(+),K(+))-ATPase and Mg(2+)-ATPase activities in homogenates of adult rat whole brain and in pure (nonmembrane bound) enzymes. DESIGN AND METHODS: Tissues were homogenized, centrifuged at 1000 xg for 10 min, and in the supernatant AChE activity and Na(+),K(+)-ATPase, Mg(2+)-ATPase activities were determined according to Ellman's method and Bowler's and Tirri's, respectively. RESULTS: After a 1-3 h CDP-choline preincubation, this substance induced a maximal stimulation of 20%-25% (p < 0.001) for AChE and 50-55% (p < 0.001) for Na(+), K(+)-ATPase, but it did not influence Mg(2+)-ATPase activity. One mM acetylcholine (ACh) resulted in an approximately 18% (p < 0.001) AChE inhibition by excess substrate in the brain homogenate, while 0. 01 mM noradrenaline did not influence Na(+),K(+)-ATPase activity. CONCLUSIONS: CDP choline can stimulate brain AChE and Na(+), K(+)-ATPase independently of ACh and noradrenaline. This enzymatic stimulation may be due to the transformation of CDP choline to membrane phophatidylcholine. The above data could explain in part the clinical effects of this substance in some neuronal disturbances. PMID- 11018687 TI - Analytical and diagnostic performance of troponin assays in patients suspicious for acute coronary syndromes. AB - BACKGROUND: The controversy whether there is a clinically significant difference between troponin T (cTnT) and troponin I (cTnI) in regard to predictive value and cardiac specificity is still ongoing. METHODS: We evaluated enzyme-linked immunosorbent assay systems for cTnI and cTnT in patients with acute coronary syndromes and multiple control groups to define threshold values for risk stratification and compare their predictive value. RESULTS: In 312 patients with noncardiac chest pain, cTnI levels were below the detection limit of 0.2 microg/L and cTnT levels were 0.011 [0.010-0. 013] microg/L. In patients with end-stage renal failure (n = 26) and acute (n = 38) or chronic (n = 16) skeletal muscle damage, median concentrations were 0.20 [0.20-0.35], below the detection limit, and 0.20 [0.20-0.25] for cTnI, and 0.04 [0.01-0.10], 0.011 [0.005-0.025], and 0.032 [0.009-0.054] microg/L for cTnT. In patients with acute coronary syndromes (n = 1130), maximized prognostic value for 30-day outcome (death, infarction) was observed at a threshold level of 1.0 microg/L for cTnI (29.0% positive) and at 0.06 microg/L for cTnT (35. 0% positive). Significant differences in the area under-the-curve values were observed between cTnI and cTnT (0.685 vs. 0.802; p = 0. 005). For both markers, the area-under-the-curve values did not increase with the second (within 24 h after enrollment) or third (48 h) blood draw. CTnI showed a less strong association with 30-day outcome than cTnT. When cTnI was put in a logistic multiple-regression model first, cTnT did add significant information. CONCLUSION: By using the defined threshold values and the employed test systems, single testing for cTnI and cTnT within 12 h after symptom onset was appropriate for risk stratification. Despite the lower cardiac specificity for cTnT, it appears to have a stronger association with the patients' outcome, whereas, as previously shown, the ability to identify patients who benefit from treatment with a GP IIb/IIIa receptor antagonist is similar. PMID- 11018688 TI - Immunofluorometric assay of human kallikrein 6 (zyme/protease M/neurosin) and preliminary clinical applications. AB - BACKGROUND: The human kallikrein gene family has contributed the best prostatic biomarkers currently available, including prostate-specific antigen (PSA) and human glandular kallikrein 2 (hK2). Recently, new members of the human kallikrein gene family have been identified. One new member is the KLK6 gene, encoding for human kallikrein 6 (hK6), which is also known as zyme/protease M/neurosin. In this paper, we describe development of antibodies and a sensitive immunofluorometric procedure for hK6 protein. METHODS: Recombinant hK6 protein was used as immunogen to develop polyclonal antibodies in rabbits and mice. These antibodies were used to develop a sandwich-type time-resolved immunofluorometric procedure for hK6. RESULTS: The newly developed hK6 immunofluorometric assay has a detection limit of 0.5 microg/L and upper concentration range of 200 microg/L. The assay is highly specific (no detectable cross-reactivity from PSA and hK2) and was used to quantify hK6 protein in various biologic fluids. Highest concentrations of hK6 were found in milk of lactating women, cerebral spinal fluid, nipple aspirate fluid, and breast cyst fluid. hK6 was also detected in male and female serum, in the majority of seminal plasmas and in a small fraction of amniotic fluids and breast tumor cytosols. hK6 was not detectable in urine. Chromatographic studies indicated that hK6 is present in these biologic fluids in its free, 30-kDa form. CONCLUSIONS: This is the first reported sensitive immunofluorometric procedure for quantifying hK6 protein. hK6 is a secreted proteolytic enzyme that is found at high levels in cerebrospinal fluid and all breast secretions. This assay will facilitate further studies to examine the possible application of hK6 in diagnostics, including cancer and neurodegenerative disorders. PMID- 11018689 TI - Genotyping of alpha-antitrypsin in ten Croatian families. AB - OBJECTIVES: The aims of the study were to determine alpha-antitrypsin (AAT) genotype by a simple DNA-based method and to investigate the association of AAT genotype and serum AAT concentration in a group of ten families. METHODS AND RESULTS: AAT genotype was determined by PCR-RFLP and serum concentration by radial immunodiffusion in samples from each member of ten families (mother, father, and child/children). In the group of parents, five normal genotypes, Pi MM, with a normal serum AAT concentration, and fifteen Pi MZ genotypes, four of them with slightly decreased (43%-66% of the mean) AAT concentration were detected. In the group of children, particular genotypes followed the mode of inheritance. There were eight Pi MZ, three of them with slightly decreased (52% 60% of the mean) AAT concentration, and five Pi ZZ genotypes with considerably decreased (24%-45% of the mean) AAT concentration. CONCLUSIONS: PCR-RFLP is the method of choice for AAT genotyping. AAT concentration is not a reliable biochemical marker of AAT deficiency. Determination of AAT genotype in family studies allows the risk of deficient allele inheritance to be followed up and assessed. Early diagnosis of a deficient AAT genotype contributes to the success of currently widely available AAT replacement therapy. PMID- 11018690 TI - Comparison of predose vs 2-h postdose blood metabolites/cyclosporine ratios in kidney and liver transplant patients. AB - OBJECTIVES: It has recently been suggested that when adjusting doses of cyclosporine (CsA), determining its concentration in blood samples taken 2 h postdose (C(2)) is more clinically beneficial than using the predose concentration (C(0)). We determined C(0) and C(2) concentrations of CsA and their metabolites in samples taken from nine kidney and seven liver transplant patients. Similarly, the so-called metabolic ratios (MR)-metabolites to CsA parent ratios-were calculated to characterise the most suitable moment of blood sampling for obtaining a greater analytical specificity with monoclonal immunoassays. METHODS: The determination of CsA and CsA + metabolites was made using the enzyme multiplied immunotechnique and the polyclonal fluorescence polarization immunoassay Abbott TDx, respectively. RESULTS: The poor correlation between C(0) and C(2) of CsA (n = 82, r = 0.387, p < 0.001) is greatly inferior to that obtained between C(0) and C(2) of metabolites (n = 82, r = 0.912, p < 0.001). A highly significant difference (p < 0.001) was found between MR(0) values (mean 2.87 +/- 0.12, median 2.48) and MR(2) values (mean 1.73 +/- 0.09, median 1.46), although there is a good correlation between them (r = 0.878, p < 0.001). CONCLUSIONS: The extent of the positive bias (deviation) of CsA immunoassays compared with the high-performance liquid chromatography results is related to the MR values. As the MR(2) values are significantly lower than the corresponding MR(0), in practice a greater analytical specificity would be obtained with the different monoclonal immunoassays in the determination of the 2 h postdose CsA concentration than in that of trough concentration. PMID- 11018691 TI - Erythrocyte protoporphyrins in hepatitis C viral infection. AB - OBJECTIVES: Protoporphyrin is the immediate precursor of the heme molecule. Due to a spillover from hemaotopoietic tissue it is regularly found in small amounts in erythrocytes and excreted into the bile. In hereditary erythropoietic protoporphyria excess protoporphyrin accumulates and can cause severe liver damage both by crystallization and induction of oxidative stress. The aim of this investigation was to study protoporphyrin concentrations in other liver disorders. DESIGN AND METHODS: Erythrocyte protoporphyrin and zinc protoporphyrin concentrations were studied in 50 patients with chronic hepatitis C infection and various degrees of liver damage. High-performance liquid chromatographic analysis with fluorescence detection was used. RESULTS: Erythrocyte protoporphyrin was increased in 32% of the patients studied; in 12 patients up to two-fold higher than the maximum of the reference range, in 4 up to three-fold higher (median concentration 98 nmol/L, interquartile range 68-142; maximum 379 nmol/L (reference range: <125 nmol/L)). In 6 of the 10 patients in the subgroup with signs of severe liver dysfunction (decreased serum albumin and prolonged thromboplastin time, elevated serum bilirubin), protoporphyrin was elevated. Erythrocyte zinc protoporphyrin was increased in 7 cases out of all 50 studied; in these seven cases, erythrocyte protoporphyrin was also elevated (median concentration of zinc protoporphyrin in the whole study group: 232 nmol/L, interquartile range 182-342; maximum 827 nmol/L (reference range <464 nmol/L). CONCLUSIONS: Elevated erythrocyte protoporphrin levels are frequently found in patients with advanced chronic hepatitis C infection. Because protoporphyrin is well known to be hepatotoxic, these findings warrant further investigation. PMID- 11018692 TI - Measurements in calcium-supplemented athletes during and after hypokinetic and ambulatory conditions. AB - STUDY OBJECTIVE: Hypokinesia (diminished movement) induces significant calcium (Ca) changes, however, little is known about Ca deficiency during hypokinesia (HK). By using Ca supplements during and after HK, the aim of this study was to establish whether HK could contribute to Ca deficiency. DESIGN AND METHODS: Studies were done during the pre-HK period of 30 days, HK period of 364 days and post-HK period of 30 days. Forty male trained athletes aged, 25.0 +/- 7.7 yr were chosen as subjects. They were equally divided into four groups: unsupplemented ambulatory control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented ambulatory control subjects (SACS), and supplemented hypokinetic subjects (SHKS). The SHKS and SACS groups took daily 55.0 mg elemental Ca per kg body weight. Hypokinetic subjects were limited to an average walking distance of 0.7 km/day, while the control subjects were running 11.6 km/day. RESULTS: Fecal Ca, urinary Ca, and urinary phosphate (P) excretion, serum ionized calcium (Ca(I)), P and total Ca levels, intact parathyroid hormone (iPTH), 1,25 dihydroxyvitamin D (1,25(OH)(2)D(3)), fluid and food intakes, peak oxygen uptake, and physical characteristics were measured. During HK fecal Ca, urinary Ca, and urinary P excretion and serum P, Ca(I), and Ca level increased significantly (p < or = 0.01), whereas during the initial days of post-HK decreased significantly (p < or = 0.01) in the SHKS and unsupplemented hypokinetic subjects groups when compared with the SACS and UACS groups, respectively. During HK serum 1,25(OH)(2)D(3), iPTH levels, food and fluid intakes, body weight, body fat, and peak oxygen uptake decreased significantly (p < or = 0.01), while during the initial days of post-HK remained significantly (p < or = 0.01) depressed and fluid intakes increased significantly (p < or = 0.01) in the SHKS and unsupplemented hypokinetic subjects groups when compared with SACS and UACS groups, respectively. Serum, urinary and fecal Ca changed much more in the SHKS than UHKS. Serum, fecal, and urinary minerals, iPTH, 1,25(OH)(2)D(3) levels, food and fluid intakes, body weight, body fat, and peak oxygen uptake did not change markedly in the SACS and UACS groups when compared with their baseline values. CONCLUSION: It was shown that serum Ca concentration, urinary, and fecal Ca excretion increased during HK and decreased significantly during post-HK. Oral Ca supplementation did not significantly affect serum Ca levels or urinary and fecal Ca loss. It was concluded that decreased urinary and fecal Ca loss during post HK, may suggest the presence of Ca deficiency during prolonged HK. PMID- 11018693 TI - Use of tumor markers for differential diagnosis of mesothelioma and secondary pleural malignancies. AB - STUDY OBJECTIVE: The aim of the study was to assess diagnosis value of tumor markers for differential diagnosis between mesothelioma and other pleural tumors. DESIGN AND METHODS: Prospective study of 85 patients attending our hospital with malignant pleural effusion. The diagnostic approach involved routine pleurocentesis followed by pleural needle. When precise diagnosis was not achieved, thoracoscopy with pleural biopsies was performed. Carcinoembryonic antigen (CEA), hyaluronic acid, tissue polypeptide antigen and cyfra 21 to 1 were measured in serum and pleural fluid. RESULTS: By using receiver operating characteristics curves and area under curves, the best diagnostic characteristics were obtained with pleural and serum CEA concentrations. The area under the curve was larger for pleural ACE than for serum ACE. The sensitivity and specificity of a pleural CEA level exceeding 3 ng/mL for ruling out the diagnosis of mesothelioma were 100% and 77%, respectively. CONCLUSION: A CEA level above 3 ng/mL in pleural fluid eliminated the diagnosis of mesothelioma, whereas the other markers were not sufficiently discriminant. However, despite a negative predictive value of 100% at a cutoff of 3 ng/mL, CEA assay in pleural fluid only avoids a small number of diagnostic thoracoscopies. PMID- 11018694 TI - Molecular genetics of glucose-6-phosphate dehydrogenase deficiency in a population of newborns from Ivory Coast. PMID- 11018696 TI - Is there an antigenic mimicry between arteriosclerotic lesions and H. pylori antigens? PMID- 11018695 TI - Status of methylation of p16 gene in multiple myeloma: a comparative study of three methods for its detection. PMID- 11018697 TI - Relationship of oxidative stress indices to decreased LDL-cholesterol after acute myocardial infarction. PMID- 11018698 TI - Variations in T1 and T2 relaxation times of normal appearing white matter and lesions in multiple sclerosis. AB - OBJECTIVE: To investigate the variation in T1 and T2 relaxation times of normal appearing white matter (NAWM) and lesions in multiple sclerosis (MS) throughout the brain. BACKGROUND: The magnetic resonance imaging (MRI) sequence fast FLAIR (fluid attenuated inversion recovery) has demonstrated overall increased lesion detection when compared to conventional or fast spin echo (FSE) but fewer lesions in the posterior fossa and spinal cord. The reasons for this are unknown, but may be due to variations in the T1 and T2 relaxation times within NAWM and MS lesions. METHOD: Ten patients and 10 controls underwent MRI of the brain which involved FSE, fast FLAIR and the measurement of T1 and T2 relaxation times. RESULTS: Of 151 lesions analysed (22 infra-tentorial, 129 supra-tentorial), eight were missed by the fast FLAIR sequence. T1 and T2 relaxation times in normal controls were longer in the infra-tentorial, than supra-tentorial, region. Patient NAWM relaxation times were prolonged compared with control values in both regions. Lesions demonstrated longer relaxation times than either control white matter or patient NAWM in both regions, however this difference was less marked infra-tentorially. The eight posterior fossa lesions not visible on the fast FLAIR sequence were characterised by short T1 and T2 relaxation times which overlapped with the patient NAWM for both T1 and T2 and with control values for T2 relaxation times. CONCLUSION: Both lesion and NAWM relaxation time characteristics vary throughout the brain. The T1 and T2 relaxation times of infra-tentorial lesions are closer to the relaxation times of local NAWM than supra-tentorial lesions, resulting in reduced contrast between posterior fossa lesions and the background NAWM. Consequently the characteristics of some lesions overlap with those of NAWM resulting in reduced conspicuity. By utilising this information, it may be possible to optimise fast FLAIR sequences to improve infra tentorial lesion detection. PMID- 11018699 TI - Treatment of Bickerstaff's brainstem encephalitis with immune globulin. AB - Bickerstaff's brainstem encephalitis (BBE) is an immune disorder of unknown etiology. Previous reports of successful treatment of BBE involved steroids or plasma exchange. We describe a patient with BBE who demonstrated rapid clinical recovery after treatment with intravenous immune globulin. Our findings suggest that IVIg is an alternative treatment in BBE, especially when plasma exchange is difficult or contraindicated. Our patient also demonstrated pseudoretraction and pseudoptosis during clinical recovery, an observation which supports a differential immune attack on components of the third nerve or supranuclear regions, and expands our understanding of the pathophysiology of BBE. PMID- 11018700 TI - Short-term motor improvement after sub-threshold 5-Hz repetitive transcranial magnetic stimulation of the primary motor hand area in Parkinson's disease. AB - Ten unmedicated patients with Parkinson's disease received sub-threshold 5-Hz repetitive transcranial magnetic stimulation (rTMS) over the primary motor hand area (M1(HAND)) contralaterally to the more affected upper limb. Compared to a midfrontal sham-rTMS, real-rTMS over M1(HAND) was associated with a significant decrease in the global motor UPDRS score 1 h after the cessation of rTMS. This was mainly due to a clinical improvement in the arm contralaterally to the stimulated M1(HAND). These preliminary data suggest that focal rTMS of M1(HAND) may have a therapeutic potential in PD. PMID- 11018701 TI - Intermuscular and intramuscular differences in myosin heavy chain composition of the human masticatory muscles. AB - Among and within the human masticatory muscles a large number of anatomical differences exists indicating that different muscles and muscle portions are specialized for certain functions. In the present study we investigated whether such a specialization is also reflected by intermuscular and intramuscular differences in fibre type composition and fibre cross-sectional area. Fibre type compositions and fibre cross-sectional areas of masticatory muscles were determined in eight cadavers using monoclonal antibodies against myosin heavy chain (MyHC). The temporalis, masseter and pterygoid muscles could be characterized by a relatively large number of fibres containing more than one MyHC isoform (hybrid fibres). In these muscles a large number of fibres expressed MyHC-I, MyHC-fetal and MyHC-cardiac alpha. Furthermore, in these muscles type I fibres had larger cross-sectional areas than type II fibres. In contrast, the mylohyoid, geniohyoid and digastric muscle were characterized by less hybrid fibres, and by less fibres expressing MyHC-I, MyHC-fetal, and MyHC-cardiac alpha, and by more fibres expressing MyHC-IIA; the cross-sectional areas of type I and type II fibres in these muscles did not differ significantly. Compared to the masseter and pterygoid muscles, the temporalis had significantly larger fibres and a notably different fibre type composition. The mylohyoid, geniohyoid, and digastric muscles did not differ significantly in their MyHC composition and fibre cross-sectional areas. Also intramuscular differences in fibre type composition were present, i.e., a regionally higher proportion of MyHC type I fibres was found in the anterior temporalis, the deep masseter, and the anterior medial pterygoid muscle portions; furthermore, significant differences were found between the bellies of the digastric. PMID- 11018702 TI - Broad-spectrum cation channel inhibition by LOE 908 MS reduces infarct volume in vivo and postmortem in focal cerebral ischemia in the rat. AB - Cation channels conduct calcium, sodium and potassium, cations that are likely deleterious in the evolution of focal ischemic injury. We studied the effects of a novel, broad-spectrum inhibitor of several cation channels, LOE 908 MS, on acute ischemic lesion development with diffusion-weighted magnetic resonance imaging (DWI) and on cerebral perfusion with perfusion imaging (PI) in vivo and on cerebral infarct size using 2,3,5-triphenyltetrazolium chloride (TTC) staining postmortem. A total of 18 male Sprague-Dawley rats underwent 90 min of middle cerebral artery occlusion (MCAO) and were randomly and blindly assigned to either LOE 908 MS or vehicle starting 30 min after inducing focal ischemia and continuing for 4 h. Whole-brain DWI and multislice PI were done before initiation of treatment and repeated frequently for the next 3.5 h. DWI-derived lesion volume at 4 h showed a significant difference in favor of the drug treated group (P=0.03), whereas PI-derived perfusion deficit volumes did not significantly differ between the groups. The postmortem infarct volume at 24 h was significantly attenuated in the treated group in comparison to controls (P=0.0001) and neurological score was significantly better in the treated group (P<0.02). Blocking several distinct cation channels with LOE 908 MS significantly reduced infarct size and improved neurological outcome without observable adverse effects in this focal ischemia model. PMID- 11018703 TI - Dialysis-related spinal canal stenosis: a clinicopathological study on amyloid deposition and its AGE modification. AB - Three cases operated for spinal canal stenosis induced by dialysis-related amyloidosis (DRA) were investigated clinicopathologically. Cases were all-male, and had undergone hemodialysis around 20 years. In two cases, cervical plain X rays showed only minor spondylotic changes. However, magnetic resonance imaging (MRI), myelography, and computed tomography (CT) showed extradural thickness with compression on the cervical spinal cord and cauda equina. In one case cervical X rays showed typical destructive spondyloarthropathy (DSA), and MRI showed compression myelopathy. Surgical treatment on both cervical and lumbar spine in two cases and on cervical spine only in one case successfully reduced the symptoms. Extradural thickened tissue and ligament flavum obtained during surgery were studied histopathologically and immunostained by using anti-CD68, anti-beta2 microglobulin (beta2m), and anti-advanced glycation end product (AGE) antibody. Congo red stain showed diffuse or nodular amyloid deposition, and immunostaining with anti-beta2m and anti-AGE antibodies also demonstrated the same distribution pattern. Thus, beta2m-positive amyloid tissue in the extradural thickness (extradural amyloid deposition) was immunohistochemically demonstrated to be modified with AGE. Inflammatory reaction with histiocytic and giant cell infiltration was also shown around the amyloid tissues. There were CD68-positive cells, and some cells were positive for AGE and beta2m. These findings suggest that beta2m accumulation and inflammatory reaction finally promote destruction of connective tissues. MRI, CT and/or myelography are necessary for diagnosing spinal canal stenosis accompanied by DRA. In conclusion, we propose a more comprehensive concept of dialysis-related spinal canal stenosis, which includes both DSA and myeloradiculopathy induced by extradural thickness. PMID- 11018704 TI - Melatonin attenuates the changes in polyamine levels induced by systemic kainate administration in rat brains. AB - Systemically administered kainate has been demonstrated to induce neuronal damage and changes of the levels of biochemical substances related to neurotoxicity. Polyamines are thought to be important in the generation of edema and neuronal cell loss associated with various type of excitotoxicity. Melatonin exerts potent free radical scavenging, antioxidant, and neuroprotective properties. This study was designed to estimate the effect of exogenous melatonin administration on the changes of polyamine levels in rat brains after systemic administration of kainate. Kainate [10 mg/kg, intraperitoneally (i.p.)] was injected into the rats to produce excitotoxicity. Melatonin (15 mg/kg, i.p.) was administered 1 h before, immediately after, and 1 h after kainate treatment. We examined the polyamine [putrescine (PU), spermidine (SD) and spermine (SM)] levels in the cerebral cortex and hippocampus and neuronal density in the hippocampal CA1 and CA3 subsectors in brain sections. PU levels were increased 8 and 24 h after kainate treatment and the administration of melatonin attenuated these changes. Only minor changes were noted in the levels of the polyamine SD and SM after the kainate treatment. In histology, neuronal injuries in the hippocampal CA1 and CA3 subsectors were examined 3 days after kainate treatment and melatonin reduced the kainate-induced neuronal injuries. Our results show that melatonin inhibits the polyamine responses in the cerebral cortex and hippocampus following kainate induced excitotoxicity and PU may be responsible for the protective effect of melatonin against kainate-induced excitotoxicity. PMID- 11018705 TI - Sex and age at diagnosis are correlated with the HLA-DR2, DQ6 haplotype in multiple sclerosis. AB - The HLA-DR2, DQ6 (i.e., HLA-DRB1*1501, DQA1*0102, DQB1*0602) haplotype contributes to the risk of developing multiple sclerosis (MS) in Caucasoids of Northern European heritage. A correlation between the clinical expression of MS and the presence of HLA-DR2, DQ6 has, however, not convincingly been shown. In this study conventional bivariate analysis and logistic regression analysis were used to study the relationship between HLA-DR2, DQ6 and four disease variables in a cohort of 286 Norwegian MS patients from the Oslo area. Logistic regression analysis showed that HLA-DR2, DQ6 was significantly more frequent among female than male patients (P=0. 0251), and was negatively correlated with age at diagnosis regardless of sex (P=0.0254). No significant correlation was observed between HLA-DR2, DQ6 and type of disease (relapsing-remitting versus primary chronic progressive MS) or presence/absence of oligoclonal bands in the cerebrospinal fluid. PMID- 11018706 TI - Behavioural assessment of functional recovery after spinal cord hemisection in the bonnet monkey (Macaca radiata). AB - In spinal cord research, current approaches to behavioural assessment often fail in defining the exact nature of motor deficits or in evaluating the return of motor behaviour from lost functions following spinal cord injury. In addition to the assessment of gross motor behaviour, it is often appropriate to use complex tests for locomotion to evaluate the masked deficits in the evaluation of functional recovery after spinal cord injury. We designed a series of sensitive quantitative tests for reflex responses and complex locomotor behaviour in the form of a combined behavioural score (CBS) to assess the recovery of function in the Bonnet monkey (Macaca radiata). Monkeys were tested for various motor/reflex components, trained to cross different complex runways, and to walk on a treadmill bipedally. The overall performance of animal's motor behaviour and the functional status of individual limb movement during bipedal locomotion was graded and scored by the CBS. Surgical hemisection was then performed on the right side of the spinal cord at the T12-L1 level. Spinal cord hemisected animals showed a significant alteration in certain reflex responses such as grasping, extension withdrawal, and placing reflexes, which persisted through 1 year of follow-up. The spinal cord hemisected animals traversed the complex locomotor runways (Narrow beam and Grid runway) with more steps and few errors, at similar levels to control animals. These observations indicate that the various motor/reflex components and bipedal locomotor behaviour of spinal cord hemisected monkeys return to control levels gradually. These results are similar to those obtained in rat models by other investigators. These results demonstrate that the basic motor strategy and the spinal pattern generator for locomotion (SPGL) in adult monkeys for the accomplishment of complex motor tasks is similar, but not identical, to that in adult rats. This suggests that the mechanisms underlying recovery are probably similar in rats and monkeys, but that primates may take a longer duration to achieve the same functional end point. PMID- 11018707 TI - High prevalence of spinocerebellar ataxia type 1 (SCA1) in an isolated region of Japan. AB - Autosomal dominant cerebeller ataxias (ADCAs) are a heterogeneous group of neurodegenerative disorders that differ in both the clinical manifestations and modes of inheritance. At present, eight different genes causing ADCAs have been found: spinocerebeller ataxia type 1 (SCA1), SCA2, SCA3/Machado-Joseph disease (MJD), SCA6, SCA7, SCA8, SCA12 and dentatorubropallidoluysian atrophy (DRPLA). The relative prevalence of each mutation varies according to race and native place. We studied 117 unrelated ADCA families that originated from the Tohoku District in the northernmost part of Honshu Island in Japan (mainly Miyagi Prefecture in the central part of Tohoku District). The SCA1 mutation was the most frequent among the known disorders (24.8% of all such families). The relative prevalence of SCA1 in the Tohoku District is very high compared with the values already reported from other regions in the world. Because the population of this area had seldom moved, the alleles with SCA1 mutations (including alleles with an intermediate CAG repeat number) are assumed to have been present in this area for a long time. PMID- 11018708 TI - De novo expansion of a CAG repeat in a Japanese patient with sporadic Huntington's disease. AB - A 49-year-old man was admitted to our hospital complaining of dysarthria and involuntary movements of his neck and extremities. He had first begun to experience involuntary neck movements at the age of 40 and his symptoms gradually progressed thereafter. There was no family history of neurological disorders. On admission he showed memory disturbance, dysarthria, and choreic movements. The involuntary movements affected his face, neck, trunk, and extremities. MRI of the brain revealed atrophy of both the cerebral cortex and the head of the caudate nucleus. DNA samples for molecular analysis were obtained from the patient and both of his parents. In this pedigree, the father carried a premutated allele of 35 CAG repeats and transmitted an expanded allele of 43 CAG repeats to his son. Paternity and maternity were analyzed using a microsatellite marker located in a different chromosome. To our knowledge, this is the first report of a sporadic case of Huntington's disease in a non-caucasian population in which the disease prevalence is much lower than that in the caucasian population. A new mutation in the current Japanese population which shares the same mechanism as de novo mutation in Caucasians may have contributed to the frequency of HD in Japan at the present time. PMID- 11018709 TI - Fluctuating monoplegia due to venous insufficiency by spinal arachnoiditis ossificans. AB - This is the first report of a patient with venous insufficiency following compressive arachnoiditis ossificans (AO). Symptoms of fluctuating monoplegia and sensory disturbance appeared monthly, lasting several weeks each time. Spinal magnetic resonance imaging (MRI) showed high T2-weighted signal intensity in the posterior portion of the column from T11 to T12 and an intradural lesion with low T2-weighted signal intensity. Neurological function and MRI improved markedly following an operation on AO. The symptoms seen in the present case were due to posterior venous insufficiency following compressive AO. PMID- 11018710 TI - Diffusion-weighted imaging in Wallerian degeneration. AB - We report two patients displaying hyperintensities on diffusion-weighted imaging (DWI) in the area of Wallerian degeneration (WD) at 12 days after stroke. High signal intensities were more conspicuous on DWI than on T2-weighted images. Both patients showed decreased diffusion anisotropy resulting in elevated apparent diffusion coefficient in the area of WD. These patients illustrate that DWI may be useful in the detection of the early stage of WD. PMID- 11018711 TI - A 3-dimensional model building by homology of the HFE protein: molecular consequences and application to antibody development. AB - Genetic hemochromatosis (GH) is a common inherited disease of iron metabolism affecting 2-5 in 1000 individuals of European origin. A candidate gene for GH, namely HFE has been recently characterized. Structural studies of the protein product of the HFE gene are of major interest for a better understanding of the molecular physiopathology in iron overload. We have built a 3-dimensional model of the HFE protein based on congruent with40% homology of sequence identity with HLA-Aw68, another MHC class I molecule. This work presents the first 3 dimensional structure of HFE available in the public domain (http://swift.embl heidelberg.de/service/francois). The 3-dimensional characteristics of the protein complexed with the beta2-microglobulin are presented. The model has been used to predict immunogenic loops and to develop an antibody able to recognize a protein exhibiting the same molecular weight as HFE. Structural consequences of two common mutations are debated and evolutionary hypotheses are considered in the discussion of the particular biological activity of HFE. This study shows that a strategy based on homology modeling is sufficient to undertake biological investigations. PMID- 11018712 TI - A bis-cyclodextrin diselenide with glutathione peroxidase-like activity. AB - A diselenide, 2,2'-diseleno-bis-beta-cyclodextrin (2-SeCD), was synthesized to imitate the antioxidant enzyme glutathione peroxidase (GPX). The GPX mimic accepts a variety of hydroperoxides as substrates. The GPX activities, reduction of H(2)O(2), tert-butyl hydroperoxide and cumenyl hydroperoxide by glutathione, are 7.4, 4.5 and 10.2 U/micromol, respectively. In contrast to ebselen (PZ51), the diselenide displays high GPX-like activity. The reduction of hydroperoxide by glutathione in the presence of a radical trap shows that the mimic catalyzes the reaction via a non-radical mechanism. A ping-pong mechanism was observed in the steady-state kinetic studies of the 2-SeCD-catalyzed reaction. PMID- 11018714 TI - Hyperactivity and interactions of a chimeric myristoryl-ACP thioesterase from the lux system of luminescent bacteria. AB - A chimeric myristoyl-ACP thioesterase with much higher catalytic efficiency than the parental enzymes has been generated by ligating the N-terminal half of the lux-specific thioesterase (LuxD) from Photobacterium phosphoreum with the C terminal half of LuxD from Vibrio harveyi. The LuxD chimera had the same rate limiting step and specificity, but cleaved esters and thioesters over eight times faster than the native enzymes. LuxD, along with acyl-protein synthetase (LuxE) and reductase (LuxC), comprise a multienzyme complex channeling activated fatty acids into the aldehyde substrate for the bacterial bioluminescence reaction. As P. phosphoreum LuxD and LuxE modulate each of their respective activities, the effects of mixing V. harveyi and the chimeric LuxD with P. phosphoreum LuxE were investigated. The chimeric LuxD stimulated acylation of LuxE to the same extent as V. harveyi LuxD, but to a lower level than that caused by P. phosphoreum LuxD. Conversely, P. phosphoreum LuxE stimulated the thioesterase activity of V. harveyi LuxD by 30% and the chimeric LuxD by 20% while the activity of P. phosphoreum LuxD was increased by over 140%. These results show that the stimulatory effects are unrelated to the level of thioesterase activity and indicate that the carboxyl terminal region of LuxD interacts with LuxE and causes a conformational change. PMID- 11018713 TI - Alcohol-induced versus anion-induced states of alpha-chymotrypsinogen A at low pH. AB - Characterization of conformational transition and folding intermediates is central to the study of protein folding. We studied the effect of various alcohols (trifluoroethanol (TFE), butanol, propanol, ethanol and methanol) and salts (K(3)FeCN(6), Na(2)SO(4), KClO(4) and KCl) on the acid-induced state of alpha-chymotrypsinogen A, a predominantly beta-sheet protein, at pH 2.0 by near UV circular dichroism (CD), far-UV CD and 1-anilinonaphthalene-8-sulfonic acid (ANS) fluorescence measurements. Addition of alcohols led to an increase in ellipticity value at 222 nm indicating the formation of alpha-helical structure. The order of effectiveness of alcohols was shown to be TFE>butanol>propanol>ethanol>methanol. ANS fluorescence data showed a decrease in fluorescence intensity on alcohol addition, suggesting burial of hydrophobic patches. The near-UV CD spectra showed disruption of tertiary structure on alcohol addition. No change in ellipticity was observed on addition of salts at pH 2.0, whereas in the presence of 2 M urea, salts were found to induce a molten globule-like state as evident from the increases in ellipticity at 222 nm and ANS fluorescence indicating exposure of hydrophobic regions of the protein. The effectiveness in inducing the molten globule-like state, i.e. both increase in ellipticity at 222 nm and increase in ANS fluorescence, followed the order K(3)FeCN(6)>Na(2)SO(4)>KClO(4)>KCl. The loss of signal in the near-UV CD spectrum on addition of alcohols indicating disordering of tertiary structure results suggested that the decrease in ANS fluorescence intensity may be attributed to the unfolding of the ANS binding sites. The results imply that the alcohol induced state had characteristics of an unfolded structure and lies between the molten globule and the unfolded state. Characterization of such partially folded states has important implications for protein folding. PMID- 11018715 TI - The effect of quercetin, a widely distributed flavonoid in food and drink, on cytosolic aldehyde dehydrogenase: a comparison with the effect of diethylstilboestrol. AB - Quercetin is a flavonoid found in red wine and many other dietary sources. Observations concerning the state of ionisation and the stability of the compound over a range of pH are presented. Quercetin is a potent inhibitor of cytosolic aldehyde dehydrogenase at physiological pH when the concentration of either the substrate or the cofactor is relatively low, but it has an activatory effect when the concentrations of substrate and cofactor are both high (1 mM). Gel filtration experiments show that quercetin binds very tightly to the enzyme under conditions where the compound is neutral and when it is ionised. The binding is less in the presence of NAD(+). Quercetin cuts down the ability of the resorufin anion to bind to the enzyme. The observations are explained by a model in which quercetin binds competitively to both the coenzyme-binding site and the aldehyde-binding site; binding in the latter location, when the enzyme is in the form of the E NADH complex, accounts for the activation. The effects of quercetin are significantly different in some respects from those of diethylstilboestrol; this is explained by the latter being able to bind to the aldehyde site but not the NAD(+) site. The possibility that quercetin may affect aldehyde dehydrogenase in vivo is discussed. PMID- 11018716 TI - Maillard reactions by alpha-oxoaldehydes: detection of glyoxal-modified proteins. AB - Proteins can be chemically modified by sugars by glycation, or the Maillard reaction. The Maillard reaction produces irreversible adducts on proteins that are collectively known as advanced glycation end products, or AGEs. Recent studies indicate that several alpha-dicarbonyl compounds, including glyoxal (GXL), are precursors of AGEs in vivo. We developed antibodies against a GXL modified protein (GXL-AGE) and purified a mixture of GXL-AGE-specific antibodies by chromatography on GXL-modified bovine serum albumin (BSA-GXL) coupled to EAH Sepharose. This preparation was then processed on a human serum albumin carboxymethyllysine (HSA-CML)-NHS-Sepharose to remove CML-specific antibodies. We used the resulting purified antibody in a competitive ELISA to probe GXL-AGEs in vitro and in vivo. We found increasingly greater antibody binding with increasing concentrations of GXL-modified BSA, but the antibody failed to react with either free CML or protein-bound CML. Incubation experiments with BSA revealed that glyceraldehyde, ribose and threose could be precursors of GXL-AGEs as well. Experiments in which GXL was incubated with N-alpha-acetyl amino acids showed that the antibody reacts mostly with lysine modifications. The GXL-derived lysine lysine crosslinking structure, GOLD was found to be one of the antigenic epitopes for the antibody. Analysis of human plasma proteins revealed significantly higher levels of GXL-AGE antigens in type II diabetic subjects compared with normal controls (P<0.0001). We also found GXL-AGEs in human lens proteins. Bovine aortic endothelial cells cultured for 7 days with 30 mM glucose did not accumulate intracellular GXL-AGEs. These studies underscore the importance of GXL for extracellular AGE formation (except in lens where it is likely to be formed intracellularly) and suggest that changes associated with age and diabetes might be prevented by alteration of GXL-AGE formation. PMID- 11018717 TI - Biphasic kinetic behavior of rat cytochrome P-4501A1-dependent monooxygenation in recombinant yeast microsomes. AB - Rat cytochrome P-4501A1-dependent monooxygenase activities were examined in detail using recombinant yeast microsomes containing rat cytochrome P-4501A1 and yeast NADPH-P-450 reductase. On 7-ethoxycoumarin, which is one of the most popular substrates of P-4501A1, the relationship between the initial velocity (v) and the substrate concentration ([S]) exhibited non-linear Michaelis-Menten kinetics. Hanes-Woolf plots ([S]/v vs. [S]) clearly showed a biphasic kinetic behavior. Aminopyrine N-demethylation also showed a biphasic kinetics. The regression analyses on the basis of the two-substrate binding model proposed by Korzekwa et al. (Biochemistry 37 (1998) 4137-4147) strongly suggest the presence of the two substrate-binding sites in P-4501A1 molecules for those substrates. An Arrhenius plot with high 7-ethoxycoumarin concentration showed a breakpoint at around 28 degrees C probably due to the change of the rate-limiting step of P 4501A1-dependent 7-ethoxycoumarin O-deethylation. However, the addition of 30% glycerol to the reaction mixture prevented observation of the breakpoint. The methanol used as a solvent of 7-ethoxycoumarin was found to be a non-competitive inhibitor. Based on the inhibition kinetics, the real V(max) value in the absence of methanol was calculated. These results strongly suggest that the recombinant yeast microsomal membrane containing a single P-450 isoform and yeast NADPH-P-450 reductase is quite useful for kinetic studies on P-450-dependent monooxygenation including an exact evaluation of inhibitory effects of organic solvents. PMID- 11018718 TI - Steady state and picosecond time-resolved fluorescence studies on native, desulpho and deflavo xanthine oxidase. AB - Steady state and time-resolved fluorescence studies on native, desulpho and deflavo xanthine oxidase (XO) have been carried out to investigate the conformational changes associated with the replacement of the molybdenum double bonded sulphur by oxygen and the removal of the flavin adenine dinucleotide (FAD). The steady state quenching experiments of the intrinsic tryptophan residues of the enzyme show that all the nine tryptophans are accessible to neutral quencher, acrylamide, in the native as well as desulpho and deflavo enzymes. However, the number of the tryptophan residues accessible to the ionic quenchers, potassium iodide and cesium chloride, increases upon removal of the FAD centre from the enzyme. This indicates that two tryptophan residues move out from the core of the enzyme to the solvent upon the removal of the FAD. The time resolved fluorescence studies were carried out on the native, desulpho and deflavo XO by means of the time-correlated single photon counting technique, and the data were analysed by discrete exponential and maximum entropy methods. The results show that the fluorescence decay curve fitted best to a three-exponential model with lifetimes tau(1)=0.4, tau(2)=1.4 and tau(3)=3.0 ns for the native and desulpho XO, and tau(1)=0.7, tau(2)=1.7 and tau(3)=4.8 ns for the deflavo XO. The replacement of the molybdenum double bonded sulphur by oxygen in the desulpho enzyme does not cause any significant change of the lifetime components. However, removal of the FAD centre causes a significant change in the shortest and longest lifetime components indicating a conformational change in the deflavo XO possibly in the flavin domain. Decay-associated emission spectra at various emission wavelengths have been used to determine the origin of the lifetimes. The results show that tau(1) and tau(3) of the native and desulpho XO originate from the tryptophan residues which are completely or partially accessible to the solvent but tau(2) corresponds to those residues which are buried in the core of the enzyme and not exposed to the solvent. For deflavo enzyme, tau(2) is red shifted compared to the native enzyme indicating the movement of tryptophan residues from the core of the enzyme to the solvents. PMID- 11018719 TI - High concentrations of D-glyceraldehyde-3-phosphate dehydrogenase stabilize the enzyme against denaturation by low concentrations of GuHCl. AB - It is known that denaturation of D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) in low concentrations of GuHCl, around 0.5 M, at 25 degrees C, leads first to a burst phase drop of activity, followed by slow unfolding with further loss of enzyme activity and aggregation. However, GAPDH at higher concentrations does not increase the aggregation in the slow phase as would be expected but decreases both the inactivation and aggregation of the enzyme instead. It seems that GAPDH at high concentrations protects the enzyme against GuHCl-denaturation. This protection is not a general effect of GuHCl binding by increased protein concentration but specific for GAPDH, as either bovine serum albumin or alpha-lactalbumin does not show any protection at similar concentrations. It is proposed that dissociation of tetrameric GAPDH into dimers in the early phase of denaturation in dilute GuHCl is reversible and further unfolding of the dimer to an aggregation prone species is irreversible and rate limiting for the unfolding process. High concentrations of the enzyme shift the equilibrium towards the tetramer thus decrease the aggregation of GAPDH in dilute GuHCl. PMID- 11018720 TI - Overproduction of beta-1,6-glucanase in Trichoderma harzianum is controlled by extracellular acidic proteases and pH. AB - To produce high amounts of extracellular endo-beta-1,6-glucanase, we overexpressed the gene bgn16.2 from Trichoderma harzianum under the control of the pyruvate kinase gene promoter (pki) of T. reesei. Transcription of bgn16.2 gene increased under most conditions but not extracellular beta-1,6-glucanase levels. Relationship of extracellular BGN16.2 protein and presence of proteases was studied in order to maximize production. After changing the carbon and nitrogen sources and buffering the culture media at different pHs, four major proteases, the acidic ones being pH-regulated, were detected. Overexpression of BGN16.2 at low pH resulted in BGN16.2 degradation, due to the induction of aspartyl proteases and to instability at pH below 3. Maximal overproduction of BGN16.2 albeit pure was achieved in buffered medium, where pH-induced aspartyl proteases were absent or when some nitrogen sources, such as yeast extract, peptone or casein were substrate for these proteases. PMID- 11018721 TI - Characterization of the ubiquitin-specific protease activity of the mouse/human Unp/Unph oncoprotein. AB - The ubiquitin-specific proteases (Ubps) are a family of largely dissimilar enzymes with two major conserved sequence regions, containing either a conserved cysteine residue or two conserved histidine residues, respectively. The murine Unp oncoprotein and its human homologue, Unph, both contain regions similar to the conserved Cys and His boxes common to all the Ubps. In this study we show that Unp and Unph are active deubiquitinating enzymes, being able to cleave ubiquitin from both natural and engineered linear ubiquitin-protein fusions, including the polyubiquitin precursor. Mutation of the conserved Unp Cys and His residues abolishes this activity, and identifies the likely His residue in the catalytic triad. Unp is tumorigenic when overexpressed in mice, leading to the suggestion that Unp may play a role in the regulation of ubiquitin-dependent protein degradation. We have demonstrated here that the high-level expression of Unp in yeast does not disrupt the degradation of the N-end rule substrate Tyr beta-galactosidase (betagal), the non-N-end rule substrate ubiquitin-Pro-betagal, or the degradation of abnormal, canavanine-containing proteins. These data suggest that Unp is not a general modulator of ubiquitin-dependent proteolysis. However, Unp may have a role in the regulation of the degradation of a specific, as yet undescribed, substrate(s). PMID- 11018722 TI - Suppression of urokinase-type plasminogen activator expression from human ovarian cancer cells by urinary trypsin inhibitor. AB - Urinary trypsin inhibitor (UTI), a Kunitz-type protease inhibitor, efficiently inhibits tumor cell invasion and metastasis. We examined the effect of UTI on urokinase-type plasminogen activator (uPA) expression in ovarian cancer cell lines, HOC-I and HRA. By Northern blot, Western blot, ELISA, and zymographic analyses, we demonstrated that UTI inhibited the expression of uPA mRNA and protein in these cells in a time- and dose-dependent manner, independent of whether induction was triggered by phorbol ester. Monoclonal antibody 4G12, which inhibits UTI binding to the cells, produced a dose-dependent abrogation in UTI mediated down-regulation of uPA expression. These data suggest that UTI significantly down-regulates tumor cell uPA mRNA expression and protein secretion, and that UTI binding to the cells is necessary to exert the UTI's action. PMID- 11018723 TI - Substitutions of surface amino acid residues of cutinase probed by aqueous two phase partitioning. AB - The surface properties of a protein are often crucial for recognition and interaction with other molecules. Important functional residues can be identified by mutational analysis. There is a need for rapid methods to study protein surfaces and surface changes due to mutations. Partitioning in aqueous two-phase systems has the potential to be used in this respect since protein partitioning depends on the surface properties of the protein. The influence of surface exposed amino acid residues in protein partitioning has been studied with cutinase variants, which differed in one or several amino acid residues as a result of site-directed mutagenesis. The solvent accessibility of the mutated residues was determined with a computer program, Graphical Representation and Analysis of Surface Properties. The aqueous two-phase system was composed of dextran and a random copolymer of ethylene oxide and propylene oxide. It was shown, for the first time, to what extent surface-exposed amino acid residues influence the partition coefficient in an aqueous two-phase system. The effect on partitioning could be described only taking into account solvent accessibility and type of residue substitution. The results demonstrate that the system can be used to detect conformational changes in mutant proteins since the expected effect on partitioning due to a mutation can be calculated. The aqueous two-phase system used here does indeed provide a rapid and convenient method to study protein surfaces and slight surface changes due to mutations. PMID- 11018724 TI - Primary structure of human hepatocellular carcinoma-associated aldehyde dehydrogenase. AB - Tumor-associated aldehyde dehydrogenase (T-ALDH) is strongly expressed in hepatocellular carcinoma (HCC) but undetectable in normal liver. In the present study, this enzyme from human HCC, HCC T-ALDH, was purified and the partial amino acid sequences (384 residues) determined by direct protein sequencing matched the amino acid sequence (453 residues) deduced from cloned HCC T-ALDH cDNAs with an open reading frame. The coding sequences of HCC T-ALDH cDNA, human stomach ALDH3A1 cDNA [Hsu et al., J. Biol. Chem. 267 (1992) 3030-3037] and human squamous cell carcinoma (SCC) T-ALDH cDNA (Schuuring et al., GenBank I.D. M74542) matched one another except for discrepancies at four positions, with consequent P12R, I27F and S134A substitutions. R and A were found in HCC and SCC T-ALDHs, whereas P and S were present in stomach ALDH3A1. To confirm that these discrepancies would have general occurrence, coding sequences of HCC T-ALDH cDNAs from six patients and stomach ALDH3A1 cDNAs from two individuals were examined and all were found to encode ALDH3A1 having R, I and A at protein positions 12, 27 and 134, respectively, indicating HCC T-ALDH to be variant ALDH3A1 which is common in human stomach tissues. PMID- 11018725 TI - Three truncated forms of serum albumin associated with pancreatic pseudocyst. AB - Plasma from a patient with chronic pancreatic pseudocyst showed an additional more negative albumin band (18%) on agarose gel electrophoresis. Both components bound (63)Ni(2+), indicating intact N-terminals; however, electrospray ionisation analysis of the intact proteins showed the mass of more negative albumin was 1254 Da less than the control and that the apparently normal band was 112 Da less. Reverse phase mapping and mass analysis of CNBr peptides showed three proteolytically modified forms of the C-terminal peptide indicating that some 81% of the albumin molecules lacked the C-terminal Leu residue, that 18% lacked the C terminal KKLVAASQAALGL and that approximately 1% lacked the QAALGL sequence. These findings were further verified by tryptic mapping of the aberrant CNBr peptides. The truncations probably result from exposure of the albumin to 'leaking' pancreatic endo and exoproteases. During less acute phases of the disease, the 13 and 6 residue truncated forms together decreased to less than 1%, while the des-Leu(585) form made up the balance; no normal albumin was detected. This suggested that the des-Leu(585) form might be present at low levels in the plasma of normal individuals and CNBr mapping confirmed that it constituted 4-15% of the albumin from normal plasma. PMID- 11018726 TI - The active-site residue tyr-175 in human glyoxalase II contributes to binding of glutathione derivatives. AB - Tyrosine-175 located in the active site of human glyoxalase II was replaced by phenylalanine in order to study the contribution of this residue to catalysis. The mutation had a marginal effect on the k(cat) value determined using S-D lactoylglutathione as substrate. However, the Y175F mutant had an 8-fold higher K(m) value than the wild-type enzyme. The competitive inhibitor S-(N-hydroxy-N bromophenylcarbamoyl)glutathione had a 30-fold higher K(i) value towards the mutant, than that of the wild-type. Pre-equilibrium fluorescence studies with the inhibitor showed that this was due to a significantly increased off-rate for the mutant enzyme. The phenolic hydroxyl group of tyrosine-175 is within hydrogen bonding distance of the amide nitrogen of the glycine in the glutathione moiety and the present study shows that this interaction makes a significant contribution to the binding of the active-site ligand. PMID- 11018727 TI - Dynamics of a mobile loop at the active site of Escherichia coli asparaginase. AB - Asparaginase II from Escherichia coli is well-known member of the bacterial class II amidohydrolases. Enzymes of this family utilize a peculiar catalytic mechanism in which a pair of threonine residues play pivotal roles. Another common feature is a mobile surface loop that closes over the active site when the substrates is bound. We have studied the motion of the loop by stopped-flow experiments using the fluorescence of tryptophan residues as the spectroscopic probe. With wild type enzyme the fluorescence of the only tryptophan, W66, was monitored. Here asparagine induced a rapid closure of the loop. The rate constants of the process (100-150 s(-1) at 4 degrees C) were considerably higher than those of the rate limiting catalytic step. A more selective spectroscopic probe was generated by replacing W66 with tyrosine and Y25, a component of the loop, with tryptophan. In the resulting enzyme variant, k(cat) and the rate of loop movement were reduced by factors of 10(2) and >10(3), respectively, while substrate binding was unaffected. This indicates that the presence of tyrosine in position 25 is essential for both loop closure and catalysis. Numerical simulations of the observed transients are consistent with a model where loop closure is an absolute prerequisite for substrate turnover. PMID- 11018728 TI - A computational study of ion binding and protonation states in the KcsA potassium channel. AB - We report results from microscopic molecular dynamics and free energy perturbation simulations of the KcsA potassium channel based on its experimental atomic structure. Conformational properties of selected amino acid residues as well as equilibrium positions of K(+) ions inside the selectivity filter and the internal water cavity are examined. Positions three and four (counting from the extracellular site) in the experimental structure correspond to distinctly separate binding sites for K(+) ions inside the selectivity filter. The protonation states of Glu71 and Asp80, which are close to each other and to the selectivity filter, as well as K(+) binding energies are determined using free energy perturbation calculations. The Glu71 residue which is buried inside a protein cavity is found to be most stable in the neutral form while the solvent exposed Asp80 is ionized. The channel altogether exothermically binds up to three ions, where two of them are located inside the selectivity filter and one in the internal water cavity. Ion permeation mechanisms are discussed in relation to these results. PMID- 11018729 TI - Kinetic characterization of Desulfovibrio gigas hydrogenase upon selective chemical modification of amino acid groups as a tool for structure-function relationships. AB - The effect of amino acid residues modification of Desulfovibrio gigas hydrogenase on different activity assays is reported. The first method consisted in the modification of glutamic and aspartic acid residues of the enzyme with ethylenediamine in order to change the polarity of certain regions of the protein surface. The second method consisted in the modification of histidine residues with a Ru complex in order to change the acid-base properties of the histidine residues. The implication of these modifications in the enzyme kinetics has been studied by measuring in parallel the activities of para/ortho hydrogen conversion, deuterium/hydrogen exchange and dyes reduction with hydrogen. Our experimental data support some hypothesis based on the three-dimensional structure of this enzyme: (a) electrostactic interactions between the hydrogenase and the redox partner play an essential role in the kinetics; (b) the histidine ligand and the surrounding acidic residues of the distal [4Fe4S] cluster form the recognition site of the redox partner of the hydrogenase; and (c) histidine residues are involved in the hydron transfer pathway of the hydrogenase. PMID- 11018730 TI - Fluorescence and nucleic acid binding properties of the human T-cell leukemia virus-type 1 nucleocapsid protein. AB - We used intrinsic tryptophan fluorescence to study the nucleocapsid protein from human T-cell leukemia virus-type one, HTLV-1 p15, an 85-amino-acid protein with two Trp-containing zinc-finger motifs. Fluorescence spectra suggested an interaction between the two zinc fingers and another interaction involving the C terminal tail and the zinc fingers. Titrations with nucleic acid revealed similar, sub-micromolar affinity for poly(dT) and poly(U) in 1 mM sodium phosphate, pH 7. Double-stranded DNA bound an order of magnitude weaker, suggesting helix-destabilizing activity. Base preference of p15 was T approximately U>I approximately C approximately G>A; affinity spanned about one order of magnitude. HTLV-1 p15 bound weaker and with less variation than reported values for either human or simian immunodeficiency virus homologues. The low affinity of p15 for nonspecific nucleic acids distinguishes it from other nucleocapsid proteins, and may suggest its involvement in additional steps of the virus life cycle other than RNA packaging. PMID- 11018731 TI - The spatio-temporal dynamics of a post-vaccination resurgence of rabies in foxes and emergency vaccination planning. AB - We used a simulation model to study the spatio-temporal dynamics of a potential rabies outbreak in an immunized fox population after the termination of a long term, large-scale vaccination program with two campaigns per year one in spring and one in autumn. The 'worst-case' scenario of rabies resurgence occurs if rabies has persisted at a low prevalence despite control and has remained undetected by a customary surveillance program or if infected individuals invade to the control area. Even if the termination of a vaccination program entails such a risk of a subsequent new outbreak, prolonged vaccination of a wild host population is expensive and the declining cost-benefit ratio over time eventually makes it uneconomic. Based on the knowledge of the spatio-temporal dynamics of a potential new outbreak gained from our modelling study, we suggest "terminating but observing" to be an appropriate strategy. Simulating the decline of population immunity without revaccination, we found that a new outbreak of rabies should be detected by customary surveillance programs within two years after the termination of the control. The time until detection does not depend on whether vaccination was terminated within the fourth, fifth or sixth years of repeated biannual campaigns. But it is faster if the program was completed with an autumn campaign (because next-year dispersal then occurs after a noticeable decrease in population immunity). Finally, if a rabid fox is detected after terminating vaccination, we determine a rule for defining a circular hazard area based on the simulated spatial spread of rabies. The radius of this area should be increased with the time since the last vaccination campaign. The trade-off between the number of foxes potentially missed by the emergency treatment and the cost for the emergency measures in an enlarged hazard area was found. PMID- 11018732 TI - Simulated economic consequences of foot-and-mouth disease epidemics and their public control in France. AB - The efficient management of foot-and-mouth disease (FMD) epidemics in France was examined through a simulation model which combines epidemiological and economic modules. From the reactions of the importing countries in terms of the products subject to import bans and the regionalization commitments, the economic module assesses the financial consequences of FMD outbreaks borne not only by the breeding sector but also by the other economic sectors on regional and national levels. Among the control options for FMD, the strategy of stamping out infected herds and dangerous in-contact herds most often contributes to reducing the economic consequences of FMD epidemics. Implementing a campaign of emergency vaccination is socially optimal if the additional export losses associated with the delay of slaughtering the vaccinated animals are offset by the gains of reducing the duration of the FMD epidemic. The importance of reducing as much as possible the total duration of the import bans is stressed by the estimated cost of an extra week of import bans. The optimal control strategy was unaffected by the introduction of stochastic parameters. PMID- 11018733 TI - A hierarchical trend model for bovine virus diarrhoea virus (BVDV) sero conversion in Norwegian dairy herds from 1993 through 1997. AB - The Norwegian bovine virus diarrhoea (BVD) control-and-eradication program performed annual testing of the national cattle population since its first screening in January 1993. A bulk-tank milk ELISA antibody test was used for the initial screening of dairy herds. Based on the annual bulk-tank milk-test result, a binary variable denoting herd sero-conversion (a surrogate measure for incidence) was created. The count of herds with sero-conversion in each community was regressed on year and the initial herd-sero-prevalence for each community in a Poisson hierarchical trend model - modelling the risk of sero-conversion. By using this modelling approach, estimates of trend specific for each hierarchical level of organisation were included in the trend model (community, veterinary district and county) could be estimated. The main BVD trend showed a steadily declining sero-conversion risk. The communities in the highest herd-sero prevalence quartile in 1993 continued to have the highest sero-conversion risk throughout the study period--decreasing from an average predicted sero-conversion risk in 1993 of 0.12 (95% CI; 0.10, 0.13) to 0.02 (0.007, 0.04) in 1997. There was an expressed variation in the level of sero-conversion for all the three hierarchical levels, but the trend only varied at the lowest level (community). PMID- 11018734 TI - Seroprevalence of Trypanosoma evansi in dromedaries (Camelus dromedarius) from the Canary Islands (Spain) using an antibody Ab-ELISA. AB - After the description in Grand Canary Island of a case of dromedary trypanosomosis caused by Trypanosoma evansi in 1998, an indirect enzyme immunoassay for the detection of specific anti-T. evansi IgG (Ab-ELISA) was used to assess the seroprevalence of this disease on the Canary Islands. Seroprevalence was 9.0% in the four studied islands (Gran Canaria, Tenerife, Lanzarote and Fuerteventura), varying from 10.0 to 7.5% by island (not significantly different). Prevalence using Ab-ELISA was higher than that observed when a parasitological method (microscopic observation of blood smears) was used (1.3%). PMID- 11018735 TI - Modelling the spread of a viral infection in equine populations managed in Thoroughbred racehorse training yards. AB - A Monte Carlo model that simulates the management life cycle of a horse population in training on a Thoroughbred flat racing yard (i.e. stable) was developed for computer implementation. Each horse was characterised by several state variables. Discrete events at the horse level were triggered stochastically to reflect uncertainty about some input assumptions and heterogeneity of the horse population in a particular yard. This mathematical model was subsequently used to mimic the spread of equine influenza (EI) within a typical yard following the introduction of one or several infectious horses. Different scenarios were simulated to demonstrate the value of strategies for preventing outbreaks of EI. Under typical UK management conditions and vaccination protocols, the model showed that EI would propagate and that the timing of vaccination in connection with the racing season and the arrival of new horses was a critical factor. The predicted outcomes (based on published characteristics of one EI vaccine) suggested that vaccination in mid-December with boosters in June and September was a viable and successful strategy in preventing the spread of EI in a training establishment. PMID- 11018736 TI - Relationships between bulk-tank antibodies to Ostertagia ostertagi and herd management practices and measures of milk production in Nova Scotia dairy herds. AB - Monthly bulk-tank milk samples were obtained from 415 Nova Scotia dairy herds in each of the months of July-September 1998 and tested using an indirect microtitre ELISA against a crude saline-extract, whole-worm Ostertagia ostertagi antigen. ELISA results (optical densities (ODs)) were consistent across months (r=0.85) but there was considerable variation among herds. A questionnaire was sent by mail to all producers; information on management factors that would potentially influence parasite burdens in the herds was obtained from 239 farms. Data on annual milk production, summer milk production (July-September) and seasonal decline in milk production were obtained from the Animal Productivity and Health Information Network (APHIN) database. Associations between management practices and ODs, and between ODs and milk-production parameters were studied. Some management practices known to be associated with parasite burdens had expected directions of association with the ODs, giving supporting evidence that the ELISA is a reasonable measure of parasite burden. Most notably, ODs were increased with greater exposure of heifers or milking cows to pasture. ODs were not associated with either annual milk production or seasonal decline in milk production. However, there was a substantial relationship between the herd OD value and the level of milk production during the summer. An increase in the OD from 0.58 to 0.83 (the interquartile range of ODs) was associated with a reduction in production of 1.25kg/cow/day. PMID- 11018737 TI - A retrospective evaluation of a Bovine Herpesvirus-1 (BHV-1) antibody ELISA on bulk-tank milk samples for classification of the BHV-1 status of Danish dairy herds. AB - Bulk-tank milk samples analysed in a Bovine Herpesvirus-1 (BHV-1) blocking ELISA are still in use in the Danish BHV-1 programme as a tool to classify dairy herds as BHV-1 infected or BHV-1 free herds. In this retrospective study, we used data from the Danish BHV-1 eradication campaign to evaluate performance characteristics of the BHV-1 blocking ELISA in 1039 BHV-1-seropositive and 502 repeatedly BHV-1-negative dairy herds using the results of blood testing of the individual animals as the true infection status. At a cut-off value of 30% blocking reaction, the herd-level relative sensitivity and relative specificity were 82 and 100%, respectively. The herd-level relative sensitivity depended on the within-herd prevalence of seropositive cows and the cut-off value in the assay, but not on the time interval (up to 90 days) between the collection of the bulk-tank milk sample and the individual serum samples. The BHV-1 blocking ELISA on bulk-tank milk could detect seropositive herds (few), with prevalence proportions as low as one seropositive cow out of 70 cows. PMID- 11018738 TI - A space-time cluster investigation of an outbreak of acute respiratory disease in Norwegian cattle herds. AB - Outbreaks of acute respiratory disease occurred in several cattle herds in the south-east regions of Norway during the winter and spring of 1995. The present study was conducted to investigate the temporal and spatial dynamics of the occurrence of acute respiratory disease between January and April 1995 in the affected cattle herds, in two neighbouring counties in Norway. This was done to assess the possible role of an infectious agent in these outbreaks, as well as to investigate the possibility of transmission between neighbouring herds. During the study period, a total of 190 (19.8%) outbreaks occurred in 955 cattle herds located in the defined study area. Descriptive maps of the spatial and spatio temporal patterns of the outbreak were generated, and two different statistical tests of space-time interaction (the Knox test and k-nearest neighbour test) and a cluster-detection test (the space-time scan statistic) were used. The results of all the space-time interaction tests strongly suggested that the case herds were clustered in time as well as in space. In addition, the space-time scan statistic defined the most-likely cluster to be located within the second veterinary district (Toten where the disease occurred) during 9-15 weeks in 1995. The relative risk of a herd being infected inside the most-likely cluster compared with the surrounding study area was estimated to be 3.59 (p=0.001). The transmission rate between neighbouring farms therefore appeared to have been higher within this cluster than in the surrounding study area. This study supports the hypothesis that one common source of infection was involved in the outbreaks during the study period. PMID- 11018739 TI - An extended state-transition model for foot-and-mouth disease epidemics in France. AB - The dynamics of foot-and-mouth disease epidemics in France was examined through simulations based on an extended state-transition model. Contagion modelling depended on specific parameters: the so-called dissemination rates. Estimation of these parameters relied on a specific discrete-event simulation model. We were able to address the problem of the hidden spread of the epidemic, before the first outbreak is diagnosed. Furthermore, we took into account the silent development of the disease in affected herds before the diagnosis. The effect of control measures such as the active search for secondary outbreaks could thus be studied. We used the model to compare the development of FMD epidemics in two very different French regions and for different control strategies implemented by the animal-health authorities. These strategies gave similar results in a low herd density area, whereas in a high herd density area, the slaughter of contact herds greatly improved the stamping-out strategy. Finally, key parameters of the model were detected through a sensitivity analysis. PMID- 11018740 TI - Somatic mutations and ageing. PMID- 11018741 TI - Introduction to a new review category PMID- 11018742 TI - A review of the genetic and related effects of 1,3-butadiene in rodents and humans. AB - In this paper, the metabolism and genetic toxicity of 1,3-butadiene (BD) and its oxidative metabolites in humans and rodents is reviewed with attention to newer data that have been published since the latest evaluation of BD by the International Agency for Research on Cancer (IARC). The oxidative metabolism of BD in mice, rats and humans is compared with emphasis on the major pathways leading to the reactive intermediates 1,2-epoxy-3-butene (EB), 1,2:3, 4 diepoxybutane (DEB), and 3,4-epoxy-1,2-butanediol (EBdiol). Results from recent studies of DNA and hemoglobin adducts indicate that EBdiol may play a more significant role in the toxicity of BD than previously thought. All three metabolites are capable of reacting with macromolecules, such as DNA and hemoglobin, and have been shown to induce a variety of genotoxic effects in mice and rats as well as in human cells in vitro. DEB is clearly the most potent of these genotoxins followed by EB, which in turn is more potent than EBdiol. Studies of mutations in lacI and lacZ mice and of the Hprt mutational spectrum in rodents and humans show that mutations at G:C base pairs are critical events in the mutagenicity of BD. In-depth analyses of the mutational spectra induced by BD and/or its oxidative metabolites should help to clarify which metabolite(s) are associated with specific mutations in each animal species and which mutational events contribute to BD-induced carcinogenicity. While the quantitative relationship between exposure to BD, its genotoxicity, and the induction of cancer in occupationally exposed humans remains to be fully established, there is sufficient data currently available to demonstrate that 1,3-butadiene is a probable human carcinogen. PMID- 11018743 TI - Factors that influence the mutagenic patterns of DNA adducts from chemical carcinogens. AB - Carcinogens are generally mutagens, which is understandable given that tumor cells grow uncontrollably because they have mutations in critical genes involved in growth control. Carcinogens often induce a complex pattern of mutations (e.g., GC-->TA, GC-->AT, etc.). These mutations are thought to be initiated when a DNA polymerase encounters a carcinogen-DNA adduct during replication. In principle, mutational complexity could be due to either a collection of different adducts each inducing a single kind of mutation (Hypothesis 1a), or a single adduct inducing different kinds of mutations (Hypothesis 1b). Examples of each are discussed. Regarding Hypothesis 1b, structural factors (e.g., DNA sequence context) and biological factors (e.g., differing DNA polymerases) that can affect the pattern of adduct mutagenesis are discussed. This raises the question: how do structural and biological factors influence the pattern of adduct mutagenesis. For structural factors, three possibilities are considered: (Hypothesis 2a) a single conformation of an adduct giving rise to multiple mutations -- dNTP insertion by DNA polymerase being influenced by (e.g.) the surrounding DNA sequence context; (Hypothesis 2b) a variation on this ("dislocation mutagenesis"); or (Hypothesis 2c) a single adduct adopting multiple conformations, each capable of giving a different pattern of mutations. Hypotheses 2a, 2b and 2c can each in principle rationalize many mutational results, including how the pattern of adduct mutagenesis might be influenced by factors, such as DNA sequence context. Five lines of evidence are discussed suggesting that Hypothesis 2c can be correct for base substitution mutagenesis. For example, previous work from our laboratory was interpreted to indicate that [+ta]-B[a]P-N(2)-dG in a 5'-CGG sequence context (G115) could be trapped in a conformation giving predominantly G-->T mutations, but heating caused the adduct to equilibrate to its thermodynamic mixture of conformations, leading to a decrease in the fraction of G-->T mutations. New work is described suggesting that [+ta]-B[a]P-N(2)-dG at G115 can also be trapped predominantly in the G-->A mutational conformation, from which equilibration can also occur, leading to an increase in the fraction of G-->T mutations. Evidence is also presented that the fraction of G-->T mutations is higher when [+ta]-B[a]P-N(2)-dG at G115 is in ss DNA ( approximately 89%) vs. ds-DNA ( approximately 66%), a finding that can be rationalized if the mixture of adduct conformations is different in ss- and ds DNA. In summary, the factors affecting adduct mutagenesis are reviewed and five lines of evidence that support one hypothesis (2c: adduct conformational complexity can cause adduct mutational complexity) are discussed. PMID- 11018744 TI - Mammalian class theta GST and differential susceptibility to carcinogens: a review. AB - Glutathione S-transferases (GSTs) are an important part of the cellular detoxification system and, perhaps, evolved to protect cells against reactive oxygen metabolites. Theta is considered the most ancient among the GSTs and theta like GSTs are found in mammals, fish, insects, plants, unicellular algae, and bacteria. It is thought that an ancestral theta-gene underwent an early duplication before the divergence of fungi and animals and further duplications generated the variety of the other classes of GSTs (alpha, mu, phi, etc.). The comparison of the aminoacidic homologies among mammals suggests that a duplication of an ancient GST theta occurred before the speciation of mammals and resulted in the subunits GSTT1 and GSTT2. The ancestral GST theta has a dehalogenase activity towards several halogenated compounds, such as the dichloromethane. In fact, some aerobic and anaerobic methylotrophic bacteria can use these molecules as the sole carbon and energy source. The mammalian GST theta cannot sustain the growth of bacteria but still retains the dehalogenating activity. Therefore, although mammalian GST theta behaves as a scavenger towards electrophiles, such as epoxides, it acts also as metabolic activator for halogenated compounds, producing a variety of intermediates potentially dangerous for DNA and cells. For example, mice exposed to dichloromethane show a dose dependent incidence of cancer via the GSTT1-1 pathway. Because GSTT1-1 is polymorphic in humans, with about 20% of Caucasians and 80% of Asians lacking the enzyme, the relationship between the phenotype and the incidence of cancer has been investigated extensively in order to detect GSTT1-1-associated differential susceptibility towards endogenous or exogenous carcinogens. The lack of the enzyme is related to a slightly increased risk of cancer of the bladder, gastro intestinal tract, and for tobacco-related tumors (lung or oral cavity). More pronounced risks were found in males with the GSTT1-null genotype for brain diseases and skin basal cell carcinomas not related to sunlight exposures. Moreover, there was an increased risk of kidney and liver tumors in humans with the GSTT1-1 positive genotype following exposures to halogenated solvents. Interestingly, the liver and kidney are two organs that express the highest level of GST theta in the human body. Thus, the GSTT1-1 genotype is suspected to confer decreased or increased risk of cancer in relation to the source of exposure; in vitro studies, mostly conducted on metabolites of butadiene, confirm the protective action of GSTT1-1, whereas, thus far, experimental studies prove that the increasing risk is limited. PMID- 11018745 TI - Biological indicators of genotoxic risk and metabolic polymorphisms. AB - International scientific publications on the influence of metabolic genotypes on biological indicators of genotoxic risk in environmental or occupational exposure are reviewed. Biomarkers of exposure (substance or its metabolites in biological fluids, urinary mutagenicity, protein and DNA adducts) and of effects (chromosome aberrations (CAs), sister chromatid exchanges (SCEs), micronuclei (Mn), COMET assay, HPRT mutants) have been evaluated according to different genotypes (or phenotypes) of several activating/detoxifying metabolic activities. In less than half the studies (43 out of 95), the influence of genotype on the examined biological indicator was found, of which four report poorly reliable results (i.e., with scarce biological plausibility, because of the inconsistency of modulated effect with the type of enzymatic activity expressed). As regards urinary metabolites, the excretion of mercapturic acids (MA) is greater in subjects with high GST activity, that of 1-pyrenol and other PAH metabolites turns out to be significantly influenced by genotypes CYP1A1 or GSTM1 null, and that of exposure indicators to aromatic amines (AA) (acetylated and non acetylated metabolites) is modulated by NAT2. In benzene exposure, preliminary results suggest an increase in urinary t, t-muconic acid (t,t-MA) in subjects with some genotypes. On urinary mutagenicity of PAH-exposed subjects, the effects of genotype GSTM1 null, alone or combined with NAT2 slow are reported. When DNA adduct levels are clearly increased in PAH-exposed group (18 out of 22), 7 out of 18 studies report the influence of GSTM1 null on this biomarker, and of the five studies which also examined genotype CYP1A1, four report the influence of genotype CYP1A1, alone or in combination with GSTM1 null. A total of 25 out of 41 publications (61%) evaluating the influence of metabolic polymorphisms on biomarkers of effect (cytogenetic markers, COMET assay, HPRT mutants) do not record any increase in the indicator due to exposure to the genotoxic agents studied, confirming the scarce sensitivity of these indicators (mainly HPRT mutants, Mn, COMET assay) for assessing environmental or occupational exposure to genotoxic substances. Concluding, in determining urinary metabolites for monitoring exposure to genotoxic substances, there is sufficient evidence that genetically-based metabolic polymorphisms must be taken into account in the future. The unfavourable association for the activating/detoxifying metabolism of PAH is also confirmed as a risk factor due to the formation of PAH-DNA adducts. The clearly protective role played by GSTT1 on DEB (and/or related compound) induced sister chromatid exchanges (SCEs) should be noted. The modulating effects of genotypes on protein adduct levels in environmental and occupational exposure have not yet been documented, and most studies on the influence of genotype on biological indicators of early genotoxic effects report negative results. PMID- 11018746 TI - The spectrum of inherited mutations causing HPRT deficiency: 75 new cases and a review of 196 previously reported cases. AB - In humans, mutations in the gene encoding the purine salvage enzyme hypoxanthine guanine phosphoribosyltransferase (HPRT) are associated with a spectrum of disease that ranges from hyperuricemia alone to hyperuricemia with profound neurological and behavioral dysfunction. Previous attempts to correlate different types or locations of mutations with different elements of the disease phenotype have been limited by the relatively small numbers of available cases. The current article describes the molecular genetic basis for 75 new cases of HPRT deficiency, reviews 196 previously reported cases, and summarizes four main conclusions that may be derived from the entire database of 271 mutations. First, the mutations associated with human disease appear dispersed throughout the hprt gene, with some sites appearing to represent relative mutational hot spots. Second, genotype-phenotype correlations provide no indication that specific disease features associate with specific mutation locations. Third, cases with less severe clinical manifestations typically have mutations that are predicted to permit some degree of residual enzyme function. Fourth, the nature of the mutation provides only a rough guide for predicting phenotypic severity. Though mutation analysis does not provide precise information for predicting disease severity, it continues to provide a valuable tool for genetic counseling in terms of confirmation of diagnoses, for identifying potential carriers, and for prenatal diagnosis. PMID- 11018747 TI - Calcium confusion--is the variability in calcium response by Sertoli cells to specific hormones meaningful or simply redundant? AB - When results of more than ten different studies on hormone-induced calcium signals in Sertoli cells are taken together, a wide variety of responses emerges. The reported changes range from increased concentrations, via no response at all, to decreased calcium concentrations. Minor variations in cell isolation techniques, culture conditions, or techniques for measuring the intracellular calcium could explain some of these differences. However, erratic variations in response are also observed within research groups under very similar experimental conditions. Such 'negative' findings are mainly reported orally and do not further penetrate the scientific community. As hormone-dependent calcium responses evidently may depend very much on the context of the cells, calcium transients would appear to be unreliable bioassay principles with which to detect the primary actions of FSH and effectors such as androgens on Sertoli cells. A more important biological question is whether these sometimes opposed calcium transients are connected with a particular cellular response. To date there is no evidence for such a tight coupling in Sertoli cells, implying that, at least under in vitro conditions, calcium signals might even be redundant altogether. Such calcium variability is probably not unique to Sertoli cells, and the aim of this commentary is to promote an open debate that may help to transform the current state of 'calcium confusion' into a better understanding of the intracellular calcium language. PMID- 11018748 TI - Thyrotrophin receptor protein expression in normal and adenomatous human pituitary. AB - Thyrotrophin (TSH) synthesis and secretion is under the positive control of thyrotrophin releasing hormone and under the negative control of the thyroid hormones. However, it is hypothesised that TSH has a direct effect on the regulation of its own synthesis through an intrapituitary loop mediated by pituitary TSH receptors (TSH-R). The aim of this investigation was to study the expression of TSH-R in normal human pituitary at mRNA and protein levels, and to compare the pattern of protein expression between different pituitary adenomas. Using RT-PCR we were able to detect TSH-R mRNA in the normal pituitary, and immunohistochemical studies showed TSH-R protein expression in distinct areas of the anterior pituitary. Double immunostaining with antibodies against each of the intrapituitary hormones and S100 revealed that TSH-R protein is present in thyrotrophs and folliculostellate cells. Examination of 58 pituitary adenomas, including two clinically active and two clinically inactive thyrotroph adenomas, revealed TSH-R immunopositivity in only the two clinically inactive thyrotroph adenomas. This study shows, for the first time, the presence of TSH-R protein in the normal anterior pituitary and in a subset of thyrotroph adenomas. The expression of TSH-R in the thyrotroph and folliculostellate cell subpopulations provides preliminary evidence of a role for TSH in autocrine and paracrine regulatory pathways within the anterior pituitary gland. PMID- 11018749 TI - Eosinophils are cellular targets of the novel uteroplacental heparin-binding cytokine decidual/trophoblast prolactin-related protein. AB - The uterus and placenta of the mouse and rat produce a member of the prolactin (PRL) family referred to as decidual/trophoblast PRL-related protein (d/tPRP). This cytokine/hormone has been hypothesized to regulate decidual cell activities needed for the establishment and maintenance of gestation. An alkaline phosphatase (AP)-tagging strategy was used to identify d/tPRP target cells. AP d/tPRP bound to virtually all cells and tissues to which it was exposed, consistent with our earlier evidence that d/tPRP binds to heparin-containing molecules. Moreover, we found that co-incubation with heparin or pretreatment with heparitinase greatly decreased the binding of AP-d/tPRP to tissue sections. In addition, we observed that the AP-d/tPRP probe bound to the surface of Chinese hamster ovary (CHO) cells but not to heparan sulfate-deficient CHO-pgsD-677 cells. Potential unique non-heparin d/tPRP binding sites within mouse and rat uteroplacental tissues were identified by consecutively incubating sections with AP-d/tPRP followed by heparin. This strategy led to the identification of d/tPRP target cells associated with the uterus and the labyrinth zone of the chorioallantoic placenta. Within the uterus, d/tPRP specifically bound to eosinophils. d/tPRP-binding and eosinophil peroxidase activity were co-localized and showed similar patterns of distribution during the estrous cycle, pregnancy, and following hormonal manipulation. d/tPRP interactions with eosinophils were further demonstrated in the lung and intestine, with eosinophils isolated from the peritoneum, and in mice with generalized tissue eosinophilia. Collectively, these findings suggest that intercellular d/tPRP targeting is mediated through associations with heparin-containing molecules which help direct d/tPRP to specific interactions with eosinophils within the uterus and with the labyrinthine compartment of the chorioallantoic placenta. PMID- 11018750 TI - Octylphenol does not mimic diethylstilbestrol-induced oestrogen receptor-alpha expression in the newborn mouse uterine epithelium after prenatal exposure. AB - This study examined whether the endocrine disruptor octylphenol (OP) mimics the synthetic oestrogen diethylstilbestrol (DES) in ability to induce oestrogen receptor-alpha (ER-alpha) expression in the newborn mouse uterine epithelium after prenatal exposure. Pregnant mice were given daily s.c. injections with DES (10 or 100 microgram DES/kg maternal wt) or OP (100 or 250 mg/kg maternal wt) or with vehicle alone from day 11.5 to 16.5 of pregnancy. ER-alpha expression was evaluated on histological sections by detecting ER-alpha mRNA with the in situ hybridization technique and ER-alpha protein using immunohistochemistry. The immunostaining was quantitated using a microspectrophotometer. Oestrogen-like activity of the DES and OP batches used for in vivo exposure was confirmed in an in vitro assay based on transient gene expression of an oestrogen-dependent reporter plasmid. In mice exposed prenatally to vehicle alone, the uterine epithelium did not express either ER-alpha mRNA or protein, while both were highly expressed in the stroma. Exposure to either DES dose induced the expression of both ER-alpha mRNA and protein in the epithelium, whereas it was unchanged in the stroma. In contrast, neither OP dose induced the expression of ER-alpha mRNA or protein in the epithelium and expression was unchanged in the stroma. Our data stress the importance of in vivo studies when investigating endocrine disruptors. PMID- 11018751 TI - Progesterone induces side-branching of the ductal epithelium in the mammary glands of peripubertal mice. AB - Development of the functional secretory epithelium in the mammary gland of the female mouse requires the elongation of the anlage through the mammary fat pad to form the primary/secondary ductal network from which tertiary ductal side branches and lobuloalveoli develop. In this study we examined the hormonal requirements for the spatial development of the primary/secondary epithelial network and tertiary side-branches by quantifying ductal growth and epithelial cell proliferation in normal and hormone-treated BALB/c mice between 21 and 39 days of age. In normal mice, an allometric increase in ductal length commenced at 31 days of age and resulted in completion of the primary/secondary ductal network by 39 days of age. Concurrent with this allometric growth was a significant increase in cellular proliferation in the terminal end-buds (TEBs) of the ductal epithelium from 29 days of age, as determined by 5-bromo-2'-deoxyuridine (BrdU) incorporation. A level of cellular proliferation similar to that in the TEBs of 33-day-old control mice could be induced in the TEBs of 25-day-old mice following treatment for 1 day with estrogen (E), or progesterone (P) or both (E/P), indicating that both E and P were mitogenic for epithelial cells of the peripubertal TEBs. However, the period of allometric ductal growth in untreated mice did not correspond to an increase in serum E or P (which might have been expected during the estrous cycle). In addition, epithelial growth was not observed in mammary glands from 24-day-old mice that were cultured in vitro with E, P or E/P. In contrast to treatment with E, treatment with P promoted a dramatic increase, relative to control mice, in the number of tertiary branch points upon the primary/secondary ductal network. BrdU labeling of mammary glands from 24- 33-day-old mice pelleted with cholesterol (C), E, P or E/P confirmed the greater mitogenicity of P on the epithelial cells of the secondary/tertiary ducts as compared with C or E. Concurrent with these changes, localized progesterone receptor (PR) expression in clusters of cells in the ductal epithelium was associated with structures that histologically resembled early branch points from ductules. In conclusion, our results suggest that additional endocrine growth factor(s) other than E and P contribute to the development of the primary/secondary ductal network, and that P is responsible for the formation of tertiary side-branches in the mammary glands of mice during puberty. PMID- 11018752 TI - Role of androgens in proliferation and differentiation of mouse mammary epithelial cell line HC11. AB - Androgens have been found in mammary epithelium and in milk throughout the cycle of the mammary gland in vivo. The aim of this study was to investigate the possible role of these substances in mammary epithelial growth and differentiation in the mouse HC11 cell line. Cells were stimulated with testosterone, dihydrotestosterone, androstenedione and 5alpha-androstane 3alpha,17beta-diol at concentrations ranging between 0.3 nM and 30 nM. Cyproterone acetate or flutamide, androgen receptor antagonists, (3 microM) were used to block specific androgen effects. Proliferative effects were measured by an MTT (tetrazolium blue) conversion test and [(3)H]thymidine uptake. HC11 cells were transfected with pbetacCAT, a chimeric rat beta-casein gene promoter chloramphenicol acetyl transferase (CAT) gene construct and CAT ELISA was used to determine gene expression. RT-PCR was performed to detect androgen receptor expression. After 24, 48 and 72 h androgens significantly (P<0.05) increased proliferation. Androgen antagonists significantly (P<0.05) reduced the proliferative effects. Furthermore androgens potentiated the lactogenic effect of prolactin, insulin and dexamethasone (P<0.05). Finally, the androgen receptor gene was expressed in both proliferating and differentiated HC11 cells. These observations lead us to hypothesize an activity of this class of steroids in mammary physiology. In particular, androgens stimulate cell proliferation and beta-casein gene expression; this influence appears to be mediated by androgen receptors. PMID- 11018753 TI - Pituitary adenylate cyclase-activating polypeptide acts synergistically with relaxin in modulating ovarian cell function in rats. AB - The interactive effects of pituitary adenylate cyclase-activating polypeptide (PACAP) and relaxin on the secretion of gelatinases, involved in matrix remodeling, in ovarian theca-interstitial cells and granulosa cells, were investigated in gonadotropin-primed immature rats. The gelatinases secreted from cultured cells were analyzed using gelatin zymography and scanning densitometry. We have previously shown that relaxin stimulated the secretion of a 71 kDa gelatinase, identified as a type IV collagenase (matrix metalloproteinase 2), in rat theca-interstitial cells. This study has demonstrated that PACAP27 and PACAP38, with similar potency, dose-dependently enhanced relaxin-induced secretion of 71 kDa gelatinase, whereas PACAP alone had no effect. In rat granulosa cells, both PACAP27 and PACAP38 alone dose-dependently increased the secretion of a 63 kDa gelatinase. In addition, this study has shown that cAMP signaling pathway mediators act similarly to that of PACAP on gelatinase secretion in rat ovarian cells. Cholera toxin, forskolin and 8-bromoadenosine cAMP augmented relaxin-induced secretion of 71 kDa gelatinase in theca interstitial cells, and alone they had no effect. These mediators also increased the secretion of 63 kDa gelatinase in granulosa cells. It is well known that the increase in cellular cAMP level is associated with the morphological rounding-up phenomenon in granulosa cells. This study has shown that PACAP and cAMP pathway mediators, but not relaxin, could cause such changes in cell shape in granulosa cells as well as in theca-interstitial cells. In conclusion, this study provides original findings that PACAP acts synergistically with relaxin in stimulating the secretion of gelatinases in rat ovarian theca-interstitial cells and granulosa cells. This supports the idea that relaxin and PACAP may serve as ovarian physiological mediators of gonadotropin function in facilitating the ovulatory process. In addition, PACAP appears to act through the cAMP signaling pathway to affect biological functions in ovarian cells, whereas relaxin does not. PMID- 11018754 TI - Effects of castration and testosterone replacement on peritoneal histamine concentration and lung histamine concentration in pubertal male rats. AB - Mast cells, which are the main source of histamine, are significantly affected by sex steroids. The present study was undertaken to determine the effects of bilateral castration and testosterone replacement on peritoneal histamine concentration and lung histamine concentration in pubertal male rats (Wistar strain). Three groups of animals were used in this study: (1) untreated castrated animals, (2) castrated animals subjected to androgen replacement by injection of propionate of testosterone, and (3) intact males as a control group. Castration alone produced a dramatic reduction in peritoneal histamine concentration. In addition, androgen replacement was effective in restoring the histamine concentration to the normal value detected in the control males (P<0.05, Kruskal Wallis test). On the other hand, there was no significant variation in the lung histamine concentration between control males, untreated castrated males and castrated males that received androgen replacement (P<0.05, Kruskal-Wallis test). These results demonstrate for the first time that castration markedly reduces the peritoneum histamine concentration in pubertal male rats, and testosterone replacement prevents the decrease. Further, these procedures do not affect lung histamine concentration, demonstrating that mast cells from different tissues may respond differently to the same biological factors. PMID- 11018755 TI - Estradiol-induced mitogen-activated protein kinase (extracellular signal regulated kinase 1 and 2) activity in the frog (Rana esculenta) testis. AB - Several lines of evidence support a key role of estradiol-17beta (E(2)) in male fertility. We have used a non-mammalian vertebrate model, the frog Rana esculenta, to investigate the regulation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) activity in the testis during the annual sexual cycle and to study whether E(2 )exerts a role in spermatogenesis through the regulation of ERK1/2 activity. ERK1/2 proteins are present in the cytoplasm and nucleus of the primary and secondary spermatogonia (SPG), and in the nucleus of primary spermatocytes. The annual E(2) profile shows a progressive increase during active spermatogenesis with a peak in the month of June. In parallel, ERK1/2 are highly phosphorylated during the period of active spermatogenesis (from April to July) compared with the regressive period (September/October) and winter stasis (from November to March). E(2) treatment induces the proliferation of primary SPG, possibly via the activation of ERK1/2, and this effect is counteracted by the anti-estrogen ICI 182-780. PMID- 11018756 TI - Possible role for protein kinase B in the anti-apoptotic effect of prolactin in rat Nb2 lymphoma cells. AB - Prolactin (PRL) is a mitogen for a number of cell types and its action as a survival factor has recently been demonstrated in Nb2 lymphoma cells. However, the intracellular signalling pathways by which PRL promotes the survival of Nb2 cells is unknown. In previous studies, we have shown that PRL caused the activation of phosphatidylinositol 3-kinase (PI3-kinase) and its association with tyrosine phosphorylated fyn. Protein kinase B (PKB), a serine/threonine kinase, is now known to be a downstream component of the PI3-kinase pathway. The aim of the present study was to examine the effect of PRL on the activation of PKB and to find out whether this has any role on the PRL-induced survival of Nb2 cells. Our studies have revealed the phosphorylation and activation of PKB in PRL stimulated Nb2 cells. We have also observed, using confocal microscopy, translocation of PKB to the membrane of Nb2 cells in response to PRL. These effects were blocked by the PI3-kinase inhibitor, LY294002 (10 microgram/ml). Apoptosis was induced by the general protein kinase inhibitor, staurosporine (STS; 0.1-1 microM), the synthetic glucocorticoid, dexamethasone (Dex; 100 nM) or ionising radiation by exposing Nb2 cells to X-irradiation (IR; 10 Gy). PRL had no effect on STS-induced apoptosis. On the other hand, PRL (100 ng/ml) inhibited apoptosis induced by Dex or IR; this effect of PRL was reversed by the addition of LY294002 (10 microgram/ml). Furthermore, Western blot analysis using phosphospecific PKB antibody on lysates from PRL-treated Nb2 cells showed that phosphorylation of PKB in response to PRL was inhibited by STS (0.5 microM), but not by Dex (100 nM). These results suggest that the PI3-kinase/PKB pathway may mediate the anti-apoptotic effect of PRL in Nb2 cells. PMID- 11018757 TI - Sustained activation of Erk1/2 MAPK and cell growth suppression by the insert negative, but not the insert-positive isoform of the human calcitonin receptor. AB - Recently we reported that calcitonin (CT) induces growth arrest at the G2 stage of the cell cycle in HEK-293 cell lines expressing the most abundant, insert negative, isoform of the human CT receptor (insert -ve hCTR). The present study investigates the involvement of the MAPK signalling pathway in the anti proliferative actions of CT and compares the activity of an isoform of the hCTR that contains a 16 amino acid insert in the first putative intracellular loop (insert +ve hCTR). Comparison of HEK-293 cells stably transfected with the insert -ve or the insert +ve hCTR, showed that accumulation of cAMP and intracellular free calcium in response to CT were specific for the insert -ve receptor isoform. However, a novel acidification of the extracellular medium was mediated by both isoforms. Treatment with CT of cells expressing the insert -ve hCTR, caused a decrease in cell growth associated with an induction of p21(WAF1/CIP1). Analysis by fluorescence-activated cell scanning showed that growth inhibition was associated with an accumulation of cells in G2. CT treatment of cells expressing the insert -ve, but not insert +ve hCTR, induced the phosphorylation of Erk1/2 MAPK, which persisted for at least 72 h. Treatment of cells expressing the insert -ve hCTR with the MAPK kinase (MEK) inhibitor, PD-98059, inhibited the phosphorylation of Erk1/2 and abrogated the growth inhibitory effects of salmon CT, the accumulation of cells in G2, and the associated induction of p21(WAF1/CIP1). These data suggest that activation of Erk1/2 are downstream effectors of the insert -ve hCTR in modulating cell cycle progression. PMID- 11018758 TI - Decreased Akt kinase activity and insulin resistance in C57BL/KsJ-Leprdb/db mice. AB - Recent studies suggest that the serine/threonine kinase protein kinase B (PKB or Akt) is involved in the pathway for insulin-stimulated glucose transporter 4 (GLUT4) translocation and glucose uptake. In this study we examined the components of the Akt signaling pathway in skeletal muscle and adipose tissue in vivo from C57BL/KsJ-Lepr(db/db) mice (db/db), a model of obesity, insulin resistance, and type II diabetes. There were no changes in the protein levels of GLUT4, p85alpha, or Akt in tissues from db/db mice compared with non-diabetic littermate controls (+/+). In response to acute insulin administration, GLUT4 recruitment to the plasma membrane increased twofold in muscle and adipose tissue from +/+ mice, but was significantly reduced by 42-43% (P<0.05) in both tissues from db/db mice. Insulin increased Akt-Ser(473) phosphorylation by two- to fivefold in muscle and adipose tissue from all mice. However, in db/db mice, maximal Akt-Ser(473) phosphorylation was decreased by 32% (P<0.05) and 69% (P<0.05) in muscle and adipose tissue respectively. This decreased phosphorylation in db/db mice corresponded with a significant decrease in maximal Akt kinase activity using a glycogen synthase kinase-3 fusion protein as a substrate (P<0.05). The level of insulin-stimulated tyrosine phosphorylation of p85alpha from phosphatidylinositol 3 (PI 3)-kinase, which is upstream of Akt, was also reduced in muscle and adipose tissue from db/db mice (P<0.05); however, there was no change in extracellular signal-regulated kinase-1 or -2 phosphorylation. These data implicate decreased insulin-stimulated Akt kinase activity as an important component underlying impaired GLUT4 translocation and insulin resistance in tissues from db/db mice. However, impaired insulin signal transduction appears to be specific for the PI 3-kinase pathway of insulin signaling, while the MAP kinase pathway remained intact. PMID- 11018759 TI - Expression of an in vitro biologically active equine LH/CG without C-terminal peptide (CTP) and/or beta26-110 disulphide bridge. AB - The C-terminal region of the beta subunit of the human chorionic gonadotrophin (hCG) is implied in heterodimer stability (beta26-110 disulphide bridge), in vitro LH bioactivity (region beta102-110) and in in vivo LH bioactivity (beta CTP). Like the hCG beta, the equine eLH and eCG beta subunits, also possess a C terminal extension (CTP). But, in contrast to hCG, eLH and eCG bind to both LH and FSH receptors in species other than the horse. This allows investigation of the roles of the beta subunit C-terminal region of a eLH/CG recombinant molecule on both LH and FSH activities. To do so, the CTP was deleted and/or the beta26 110 disulphide bond was mutated and the resulting mutated beta subunits were transiently co-expressed with common alpha subunit in COS7 cells. These regions were also deleted in a betaalphaeLH/CG single chain also expressed in COS7 cells. The hormones produced were characterized by different ELISAs and in vitro LH and FSH bioassays. Mutation of the 26-110 disulphide bond and deletion of the betaCTP led to a decrease in eLH/CG heterodimer production. Double mutation promoted an additive effect on production of the heterodimer and of the corresponding tethered eLH/CG. The elimination of the beta26-110 disulphide bond in the betaalpha single chain had no effect on its production. However, neither the 26 110 disulphide bond nor the CTP mutations affected dimer stability and bioactivities of the secreted heterodimers and/or single chain molecules. Therefore, in contrast to hCG, the 26-110 S-S bond of the recombinant eLH/CG beta subunit does not seem to be essential for eLH/CG dimer stability upon secretion and expressing LH and FSH bioactivities. PMID- 11018760 TI - Induction of follicle formation in long-term cultured normal human thyroid cells treated with thyrotropin stimulates iodide uptake but not sodium/iodide symporter messenger RNA and protein expression. AB - Iodide uptake by the sodium/iodide symporter (NIS) in thyrocytes is essential for thyroid hormone production. Reduced NIS activity has been reported in thyroid diseases, including thyroid cancer and congenital hypothyroidism. The study of iodide uptake in thyrocytes has been limited by the availability of appropriate in vitro models. A new culture technique was recently developed that allows normal human thyroid primary culture cells to grow as monolayer cells and express differentiated functions for more than 3 months. We used this technique to study the effect of follicle formation and TSH on iodide uptake in these cells. Iodide uptake by the cells grown in monolayer was very low. Follicle formation was induced from monolayer cells, and electron micrographs demonstrated cell polarity in the follicles. No significant increase in iodide uptake was observed after TSH treatment of cells in monolayer or when follicle formation was induced without TSH. TSH stimulation of follicles, however, significantly increased iodide uptake ( approximately 4. 4-fold; P<0.001). Compared with iodide uptake in monolayers, the combination of follicle formation and TSH treatment stimulated iodide uptake synergistically to 12.0-fold (P<0.001). NIS messenger RNA (mRNA) and protein levels were almost the same in both monolayer cells and follicles. TSH treatment of monolayers and follicles produced significant (P<0.05) stimulation of mRNA ( approximately 4. 8- and approximately 4.3-fold respectively) and protein ( approximately 6.8- and 4.9-fold respectively). TSH stimulated NIS protein levels in both monolayer and follicles, however, stimulation of functional iodide uptake was only seen with TSH stimulation of follicles. The function of NIS may involve post-transcriptional events, such as intracellular sorting, membrane localization of NIS or another NIS regulatory factor. Polarized functions, such as iodide efflux into follicular lumina, may also contribute to the increased iodide concentration after follicle formation. PMID- 11018761 TI - Neurohypophysial hormone receptors and second messengers in trout hepatocytes. AB - Neurohypophysial hormone receptors and second messengers were studied in trout (Oncorhynchus mykiss) hepatocytes. Arginine vasotocin (AVT) and isotocin (IT) elicited a concentration-dependent inhibition of cAMP accumulation in the presence of 5x10(-8) M glucagon (maximal effect for 4.5x10(-7) M and 1.4x10(-7) M, half-maximal effect for 2.1x10(-8) M and 0.7x10(-8) M, AVT and IT respectively). The effect of glucagon was inhibited up to 90% by AVT and 80% by IT. While AVT inhibited (up to 50%) the basal cAMP production, IT had no such action. Specific V(1) or V(2) analogues (with reference to vasopressin in mammals) were used for pharmacological characterization of the type of neurohypophysial hormone receptor involved in this inhibition. The V(1) agonist [Phe(2), Orn(8)]-oxytocin inhibited the glucagon-stimulated cAMP production with a maximal effect for 6x10(-7) M and a half-maximal effect for 0.9x10(-8) M concentrations of the analogue. While the V(1) agonist reduced the glucagon stimulated cAMP level by 70%, it showed only a tendency to reduce the basal level. The V(2) agonist [deamino(1), Val(4),d -Arg(8)]-vasopressin had no effect either on basal or on glucagon-stimulated cAMP production. The V(1) antagonist [d(CH(2))(5)(1), O-Me-Tyr(2), Arg(8)]-vasopressin totally reversed the 10(-8) M AVT-induced inhibition of 5x10(-8) M glucagon-stimulated cAMP production, whereas the V(2) antagonist [d(CH(2))(5)(1),d -Ile(2), Ile(4), Arg(8), Ala(9)] vasopressin had no such effect. In this particular case, maximal and half-maximal effects of the V(1) antagonist were obtained for 2.3x10(-6) M and 1. 2x10(-6 )M respectively. Changes in intracellular calcium content were measured using the fluorescent probe FURA-2/AM. AVT and IT elicited a concentration-dependent increase in Ca(2+) accumulation. The comparison of the effect of 10(-8) M agonists versus AVT showed the following order of potency: AVT=IT>V(1) agonist>V(2) agonist. The V(1) antagonist reversed the AVT-induced Ca(2+) accumulation whereas the V(2) antagonist had no such effect. These results are taken as evidence for the presence in trout hepatocytes of neurohypophysial hormone receptors functionally close to the V(1a)-type linked to cAMP production and Ca(2+) mobilization. PMID- 11018762 TI - Alternative 5'-untranslated regions of mouse GH receptor/binding protein messenger RNA are derived from sequences adjacent to the major L2 promoter. AB - Heterogeneity of 5' untranslated region (5'UTR) sequences is a common feature of growth hormone receptor/binding protein (GHR/BP) mRNA from a number of species. Two major 5'UTR sequences (designated L1 and L2 in the mouse) have been cloned from rodents, ruminants and primates, and are known to correspond to transcripts generated from independently regulated promoters. A variable number of other 5'UTRs with diverse sequences have been cloned from rat, human and bovine tissues. To characterize alternative 5'UTR usage in mouse GHR/BP mRNA, we carried out 5' rapid amplification of cDNA ends using RNA from non-pregnant mouse liver and adipose tissue. Three novel 5'UTR sequences were obtained. Sequencing of genomic DNA revealed that exons corresponding to these three sequences are clustered within 1 kb downstream of the exon encoding 5'UTR L2, and the associated L2 promoter. The novel 5'UTRs are present at very low levels relative to the total pool of GHR/BP mRNA in liver, fat, kidney, and mammary gland as determined by ribonuclease protection assays. On the basis of these data, we propose that these 5'UTR sequences may result from the use of cryptic transcription start sites and splice donor sites under the influence of the adjacent L2 promoter/enhancer region. PMID- 11018763 TI - Production and characterization of specific antibodies for evaluation of glycated insulin in plasma and biological tissues. AB - Previous studies have shown that glycation of insulin occurs in pancreatic beta cells under conditions of hyperglycaemia and that the site of glycation is the N terminal Phe(1) of the insulin B-chain. To enable evaluation of glycated insulin in diabetes, specific antibodies were raised in rabbits and guinea-pigs by using two synthetic peptides (A: Phe-Val-Asn-Gln-His-Leu-Cys-Tyr, and B: Phe-Val-Asn Gln-His-Leu-Tyr-Lys) modified by N-terminal glycation and corresponding closely to the N-terminal sequence of the glycated human insulin B-chain. For immunization, the glycated peptides were conjugated either to keyhole limpet haemocyanin or ovalbumin using glutaraldehyde, m-maleimidobenzoyl-N hydroxysuccinimide ester or 1-ethyl-3-(3-dimethylamino propyl) carbodiimide hydrochloride. Antibody titration curves, obtained using I(125)-tyrosylated tracer prepared from glycated peptide A, revealed high-titre antisera in five groups of animals immunized for 8-28 weeks. The highest titres were observed in rabbits and guinea-pigs immunized with peptide B coupled to ovalbumin using glutaraldehyde. Under radioimmunoassay conditions, these antisera exhibited effective dose (median) (ED(50)) values for glycated insulin of 0.3-15 ng/ml and 0.9-2.5 ng/ml respectively, with negligible cross-reactivity against insulin or other islet peptides. The degree of cross-reaction with glycated proinsulin was approximately 50%. Glycated insulin in plasma of control and hydrocortisone treated diabetic rats measured using rabbit 3 antiserum (1:10 000 dilution; sensitivity <19 pg/ml) was 0. 08+/-0.01 and 1.5+/-0.6 ng/ml (P<0.01), corresponding to 4 and 16% of total circulating insulin concentration respectively. Immunocytochemistry studies of the pancreas of streptozotocin treated diabetic rats using a 1:1000 dilution of guinea-pig 2 antiserum revealed clusters of fluorescent positively stained cells in islets. These studies document the successful production of polyclonal antisera specific for glycated insulin and their usefulness in radioimmunoassays and immunocytochemistry. The demonstration of glycated insulin in plasma and islets of animal models of diabetes supports the view that glycation of insulin is involved in the pathogenesis of this disease. PMID- 11018764 TI - IGF-I inhibits apoptosis induced by serum withdrawal, but potentiates TNF-alpha induced apoptosis, in 3T3-L1 preadipocytes. AB - We have previously shown that human preadipocytes in primary culture undergo apoptosis in response to serum deprivation and addition of tumour necrosis factor alpha (TNF-alpha), and have proposed that regulation of preadipocyte apoptosis in vivo may contribute to the overall control of adipose mass. In the present study we have investigated both pro- and anti-apoptotic factors, and the signalling pathways by which they act, in murine 3T3-L1 preadipocytes. Apoptotic indices (fraction of cells undergoing apoptosis) were determined by microscopic examination of acridine orange-stained cells, fluorescence-activated cell sorting of propidium iodide-stained cells, or phase-contrast video microscopy. Murine 3T3 L1 cells were more susceptible to apoptosis than human preadipocytes. In medium containing 10% newborn calf serum, the basal apoptotic index was very low (<2%), but the number of apoptotic cells increased significantly following serum withdrawal (10% after 24 h). Addition of TNF-alpha (6 nM) stimulated apoptosis in both serum-containing and serum-free media (apoptotic indices of 12% and 20% respectively after 24 h). IGF-I inhibited by approximately 50% the apoptosis induced by serum withdrawal, but increased by 25% the apoptosis induced by TNF alpha in serum-free medium. It was shown by using specific inhibitors of lipid and protein kinases (LY294002, rapamycin, PD98059, SB203580) that both phosphoinositide 3-kinase and MAP kinase pathways contribute to the anti apoptotic action of IGF-I on serum-starved cells, while phosphoinositide 3-kinase but not MAP kinase activity is required for the paradoxical pro-apoptotic action of IGF-I in the presence of TNF-alpha. We conclude that, in addition to its previously described anti-apoptotic action, IGF-I can also be pro-apoptotic in 3T3-L1 cells in the presence of TNF-alpha, and that both the anti- and pro apoptotic effects of IGF-I require the activation of phosphoinositide 3-kinase. PMID- 11018765 TI - Functional inactivation of the IGF-I receptor delays differentiation of skeletal muscle cells. AB - Skeletal myoblasts are inherently programmed to leave the cell cycle and begin the differentiation process following removal of exogenous growth factors. Serum withdrawal results in a marked induction of IGF production which is essential for skeletal muscle differentiation in vitro. However, the potential role of the tyrosine kinase IGF-I receptor (thought to be the principal mediator of both IGF I and II signaling in skeletal muscle) in the decision of myoblasts to begin differentiation following serum withdrawal is unknown. To explore the role of the IGF-I receptor in this decision by skeletal myoblasts, we functionally inactivated endogenous IGF-I receptors in mouse C2C12 cells using a dominant negative, kinase-inactive IGF-I receptor in which the ATP-binding site lysine (K) at residue 1003 has been mutated to alanine (A). Cell lines with the greatest degree of mutant IGF-I receptor expression (A/K cells) demonstrated functional inactivation of endogenous IGF-I receptors as determined by their impaired ability to phosphorylate the principal substrate of the IGF-I receptor, IRS-1, in response to treatment with IGF-I. In addition, the proliferative response of myoblasts to IGF-I was completely abolished in A/K cells. Following withdrawal of exogenous growth factors, A/K cells demonstrated a marked delay in the induction of the gene expression of myogenin, a skeletal muscle-specific transcription factor essential for differentiation, and a subsequent delay in the induction of muscle creatine kinase activity. Delayed differentiation in A/K cells was associated with prolonged phosphorylation of the cell cycle regulatory retinoblastoma (Rb) protein; it is the un- (or hypo-) phosphorylated form of Rb which is known to promote differentiation in skeletal myoblasts. Thus, the IGF-I receptor regulates the timing of myoblast differentiation induced by serum withdrawal. The delayed differentiation of skeletal myoblasts with functionally inactive IGF-I receptors may result, at least in part, from delayed induction of myogenin gene expression and prolonged phosphorylation of the Rb protein. PMID- 11018766 TI - Inhibition of dioxin effects on bone formation in vitro by a newly described aryl hydrocarbon receptor antagonist, resveratrol. AB - Aryl hydrocarbon receptor (AhR) ligands are environmental contaminants found in cigarette smoke and other sources of air pollution. The prototypical compound is TCDD (2,3,7, 8-tetrachlorodibenzo-p-dioxin), also known as dioxin. There is an increasing body of knowledge linking cigarette smoking to osteoporosis and periodontal disease, but the direct effects of smoke-associated aryl hydrocarbons on bone are not well understood. Through the use of resveratrol (3,5,4' trihydroxystilbene), a plant antifungal compound that we have recently demonstrated to be a pure AhR antagonist, we have investigated the effects of TCDD on osteogenesis. It was postulated that TCDD would inhibit osteogenesis in bone-forming cultures and that this inhibition would be antagonized by resveratrol. We employed the chicken periosteal osteogenesis (CPO) model, which has been shown to form bone in vitro in a pattern morphologically and biochemically similar to that seen in vivo, as well as a rat stromal cell bone nodule formation model. In the CPO model, alkaline phosphatase (AP) activity was reduced by up to 50% (P<0.01 vs control) in the presence of 10(-9) M TCDD and these effects were reversed by 10(-6) M resveratrol (P<0.05 vs TCDD alone). TCDD mediated inhibition of osteogenesis was restricted primarily to the osteoblastic differentiation phase (days 0-2) as later addition did not appear to have any effects. Message levels for important bone-associated proteins (in the CPO model) such as collagen type I, osteopontin, bone sialoprotein and AP were inhibited by TCDD, an effect that was antagonized by resveratrol. Similar findings were obtained using the rat stromal bone cell line. TCDD (at concentrations as low as 10(-10)M) caused an approximately 33% reduction in AP activity, which was abrogated by 3. 5x10(-7) M resveratrol. TCDD also induced a marked reduction in mineralization ( approximately 75%) which was completely antagonized by resveratrol. These data suggest that AhR ligands inhibit osteogenesis probably through inhibition of osteodifferentiation and that this effect can be antagonized by resveratrol. Since high levels of AhR ligands are found in cigarette smoke, and further since smoking is an important risk factor in both osteoporosis and periodontal disease, it may be postulated that AhR ligands are the component of cigarette smoke linking smoking to osteoporosis and periodontal disease. If so, resveratrol could prove to be a promising preventive or therapeutic agent for smoking-related bone loss. PMID- 11018767 TI - Gene expression of the three members of hepatocyte nuclear factor-3 is differentially regulated by nutritional and hormonal factors. AB - Hepatocyte nuclear factor-3 (HNF-3) belongs to a large family of forkhead transcription factors and is made up of three members (HNF-3alpha, -3beta and 3gamma). It has been shown that HNF-3 regulates a number of metabolically important genes. However, the mechanisms underlying this regulation of HNF-3 activity by hormones and nutrition have not yet been well elucidated. In attempting to explore the regulation of gene expression of HNF-3 members by physiological status, we analyzed the effects of insulin, dexamethasone and protein malnutrition on the hepatic mRNA level of each member. Male Wistar rats were fed on a 12% casein diet, 12% gluten diet (deficient in lysine and threonine) or a protein-free diet for 1 week. The protein-free diet and gluten diet caused a 3. 7-fold increase in HNF-3g mRNA in the liver and did not affect the mRNA level of either HNF-3alpha or HNF-3beta. Daily administration of dexamethasone caused the mRNA levels of HNF-3alpha and HNF-3beta to increase (2.3 and 1.4-fold, respectively), but had no effect on the HNF-3gamma mRNA level. In diabetic rats that had been injected with streptozotocin, an elevation of the hepatic mRNA levels of HNF-3beta and HNF-3gamma was observed (1.6-and 1.9-fold, respectively). Insulin replacement in the diabetic rats decreased both mRNA levels in a dose-dependent manner. HNF-3alpha mRNA was not affected by insulin status. These results show that the genes of the three members of the HNF-3 family respond differently to hormonal and nutritional factors suggesting that the activities of HNF-3 members are regulated, at least in part, by the levels of their gene expression. PMID- 11018768 TI - Local and cortical effects of olfactory bulb lesions on trophic support and cholinergic function and their modulation by estrogen. AB - This study determined whether olfactory bulb lesions would affect trophic support to its afferent, the horizontal limb of the diagonal band of Broca (hIDBB), and if estrogen would ameliorate the effects of neural injury in this circuit. NMDA injections into the olfactory bulb resulted in neural injury as indicated by cell loss and increased glial fibrillary acidic protein immunoreactivity. Olfactory bulb lesions severely reduced BDNF expression in its afferent, the hIDBB, while NGF was only reduced in lesioned animals deprived of estrogen. In the olfactory bulb itself, lesions increased BDNF expression, but not NGF. Paradoxically, bulb lesions up-regulated both NGF and BDNF in another target of the hIDBB, the cingulate cortex. Moreover, olfactory bulb lesions affected choline uptake and ChAT activity locally, as well as in the cingulate cortex. Estrogen significantly attenuated the lesion-induced loss of choline uptake in the cingulate cortex, but not at the primary lesion site. Collectively, these results indicate that neural injury to one limb of the forebrain cholinergic system may result in collateral damage to other limbs of this system, suggesting a mechanism for the progression of neurodegenerative diseases, such as Alzheimer's disease, that involve the cholinergic system. Furthermore, these data also indicate that estrogen selectively attenuates certain lesion-induced deficits. PMID- 11018769 TI - Developmental disinhibition: turning off inhibition turns on breathing in vertebrates. AB - Development requires age-dependent changes in essential behaviors. While the mechanisms determining the developmental expression of such behavior in vertebrates remain largely unknown, a few studies have identified permissive mechanisms in which the appearance of promoting signals activates pre-established networks. Here we report a different developmental process. Specifically, we show that the neuronal substrate that produces putative lung breathing in tadpoles is formed early in development, but remains more or less inactive until metamorphosis because of suppression mediated by a GABA(B) receptor-dependent mechanism. Blocking this suppression using 2-hydroxy-saclofen, a GABA(B) receptor antagonist, results in the precocious production of the putative lung breathing motor pattern. This blocker failed to augment putative lung breaths after metamorphosis. Thus, our results suggest that loss of an inhibitory signal during development (i.e., developmental disinhibition) is responsible for the developmental expression of air breathing. PMID- 11018770 TI - Positive injury signals induce growth and prolong survival in Aplysia neurons. AB - Injury to a peripheral nerve initiates changes that can lead to regeneration of the damaged axons. How information about a distant injury is communicated to the cell body is not clear. Using the nervous system of Aplysia californica, we tested the idea that some of this information is conveyed via positive injury signals-axoplasmic proteins that are activated at the injury site and transported to the cell soma. We collected these proteins by crushing pedal nerves and then placing a ligation proximal to the ligation. The contralateral nerves were ligated as controls. Twenty h later, axoplasm was extruded from the nerve segment just distal to the ligation on the crushed nerves (cr/lig) and on the control nerves (lig). The total proteins were rhodaminated and injected into the cytoplasm of neurons in vitro to look for nuclear import. Punctate fluorescence was detected in the nucleus of all seven neurons injected with the cr/lig axoplasm. Only two of five neurons injected with lig axoplasm had any fluorescence. Equal amounts of cr/lig and lig axoplasm were then injected directly into the cell bodies of neurons maintained in vitro. The cells injected with cr/lig axoplasm exhibited renewed growth and significantly longer survival: 25.9 +/- 2.1 days (mean +/- SEM: n = 22) relative to the cells injected with lig axoplasm (15.3 +/- 1.2 days; n = 14) and to those that were not injected (12.2 +/ 1.7 days; n = 24). Fractionation of the cr/lig axoplasm indicated that different factors are responsible for growth and survival. PMID- 11018771 TI - Overexpression of NGF ameliorates ethanol neurotoxicity in the developing cerebellum. AB - Transgenic mice overexpressing NGF in the central nervous system under the control of the glial fibrillary acidic protein (GFAP) promoter were exposed to ethanol via vapor inhalation on postnatal days 4 and 5 (P4-5), the period of maximal cerebellar Purkinje cell sensitivity to ethanol. Wild-type controls were exposed in a similar manner. There were no differences in body weight or size following these procedures, but the transgenic brain weights at this age were significantly greater than wild-type controls. In the wild-type animals, a significant 33.3% ethanol-mediated loss of Purkinje cells in lobule I was detected via unbiased three-dimensional stereological counting on P5. In the GFAP NGF transgenic animals, however, the 17.6% difference in Purkinje cell number in control and ethanol-exposed animals was not significant. There was a similar difference in Purkinje cell density in both groups, which did reach statistical significance (-32.7% in wild-type ethanol-treated animals, -17% in transgenic ethanol-exposed animals). These results suggest that endogenous overexpression of neurotrophic factors, which have previously been shown to protect against ethanol neurotoxicity in culture, can serve a similar protective function in the intact animal. PMID- 11018772 TI - Drosophila endosomal proteins hook and deep orange regulate synapse size but not synaptic vesicle recycling. AB - To study the function of endosomes at synapses we analyzed the localization and function of two Drosophila endosomal proteins, Hook and Deep orange (Dor), at the larval neuromuscular junction. Hook, a negative regulator of endocytic trafficking, and Dor, a positive regulator of endocytic trafficking, are highly enriched at synapses, especially close to postsynaptic membranes. Mutations in hook (hk) and dor do not affect synaptic vesicle recycling, as assessed by electrophysiological analysis of synaptic transmission and behavioral studies of double mutants with shi(ts) mutations that alter vesicle recycling. However, hk and dor mutations alter the number of presynaptic varicosities (synapse size) in opposing ways. Synapse size is increased in hk(11) mutants and is decreased in dor(4) mutants. Double mutants for dor and hk show a dor-like phenotype. These effects on synapse size parallel known functions of Hook and Dor in endocytosis and strongly indicate a role for endocytic trafficking in the regulation of synapse size in vivo. Our observations suggest a model in which Hook and Dor function in later stages of endocytosis is essential for regulating synaptic plasma membrane composition but not synaptic vesicle recycling. PMID- 11018773 TI - Nitric oxide modulates retinal ganglion cell axon arbor remodeling in vivo. AB - Nitric oxide (NO) has been postulated to act as an activity-dependent retrograde signal that can mediate multiple aspects of synaptic plasticity during development. In the visual system, a role for NO in activity-dependent structural modification of presynaptic arbors has been proposed based on NO's ability to prune inappropriate projections and segregate axon terminals. However, evidence demonstrating that altered NO signaling does not perturb ocular dominance map formation leaves unsettled the role of NO during the in vivo refinement of visual connections. To determine whether NO modulates the structural remodeling of individual presynaptic terminal arbors in vivo we have: 1. Used NADPH-diaphorase histochemistry to determine the onset of NO synthase (NOS) expression in the Xenopus visual system. 2. Used in vivo time-lapse imaging to examine the role of NO during retinal ganglion cell (RGC) axon arborization. We show that NOS expression in the target optic tectum is developmentally regulated and localized to neurons that reside in close proximity to arborizing RGC axons. Moreover, we demonstrate that perturbations in tectal NO levels rapidly and significantly alter the dynamic branching of RGC arbors in vivo. Tectal injection of NO donors increased the addition of new branches, but not their stabilization in the long term. Tectal injection of NOS inhibitors increased the dynamic remodeling of axonal arbors by increasing branch addition and elimination and by lengthening pre-existing branches. Thus, these results indicate that altering NO signaling significantly modifies axon branch dynamics in a manner similar to altering neuronal activity levels (Cohen-Cory, 1999). Consequently, our results support a role for NO during the dynamic remodeling of axon arbors in vivo, and suggest that NO functions as an activity-dependent retrograde signal during the refinement of visual connections. PMID- 11018775 TI - Introduction to this issue: international perspectives on therapeutic jurisprudence: part II PMID- 11018774 TI - Erratum to "Growth cone form, behavior, and interactions in vivo: retinal axon pathfinding as a model" PMID- 11018776 TI - Can the criminal law ever be therapeutic? AB - This paper addresses the question of whether criminal law, especially in its sentencing and penological aspects, can be adjusted in its workings to incorporate the perspective of therapeutic jurisprudence. A conceptual framework developed by Wexler, Winick, and their colleagues is employed to provide a basis for reflection on the operation and impact of sentencing processes. Of considerable importance in this exercise is a mounting volume of evidence concerning the outcomes of work with adjudicated offenders, which illustrates the potential of psycho-educational, behavioral-cognitive skills-training, and therapeutic programs for reduction of recidivism. Such intervention programs are currently offered within the limits set by, but are not intrinsic components of, judicial decisions. The psycho-legal implications of these findings are discussed and some possibilities surveyed for tentative exploration of actively therapeutic departures within crimino-legal decision-making. PMID- 11018777 TI - From the psychiatric hospital to the community: integrating conditional release and contingency management. AB - Psychiatric hospital recidivism has been and continues to be a persistent problem in treating individuals with chronic mental illness. Conditional release, a form of involuntary outpatient commitment, has been suggested as one possible solution. Guided by therapeutic jurisprudence, this article presents a proposal about conditional release that would maximize convergence of social values and would be empirically testable. Specifically, a scientifically validated treatment intervention for individuals with chronic mental illness, contingency management, is integrated with conditional release. From this proposal, a number of empirical hypotheses and legal questions about discharging psychiatric patients are generated and discussed. PMID- 11018778 TI - Therapeutic jurisprudence in the courts. AB - Therapeutic jurisprudence is an emerging field of law and social science inquiry that explores the role of the law in fostering therapeutic or antitherapeutic outcomes. This article considers the relationship between therapeutic jurisprudence and court performance goals, examines applications of therapeutic jurisprudence in court settings, discusses the steps involved in incorporating therapeutic jurisprudence principles into the work of courts, outlines the pros and cons associated with practicing therapeutic jurisprudence primarily in specialized courts, and offers suggestions for fostering continued experimentation by courts. PMID- 11018779 TI - The South African truth and reconciliation commission as a therapeutic tool. AB - A Truth Commission is one of the institutions used in international law to investigate gross human rights violations within a specific country. In this article we examine claims that the South African Truth and Reconciliation Commission (TRC) was therapeutic. In the absence of empirical evidence, this examination will be guided by a theoretical framework that will reflect ways by which we believe international legal institutions can contribute tot he healing of the people of a country in which human rights abuses have taken place. We developed this framework with reference to the literature. Our conclusion is that the legislator's emphasis on truth, reconciliation, stability, and restorative justice enhanced the TRC's potential to promote healing, but that some features of this procedure and the enabling Act, the Promotion of National Unity and Reconciliation Act (1995), restricted its ability to be therapeutic. We conclude by looking at the role culture may have played in the success of the TRC. PMID- 11018780 TI - Maximizing therapeutic effects in treating sexual offenders in an Australian correctional system. AB - In Australia, the community response to sexual offenders is marked by uncertainty as to whether offenders should be incarcerated as punishment or provided treatment in order to reduce the likelihood of re-offense. The incarceration of sexual offenders results in particular management, ethical, and political issues. Nevertheless incarceration can provide leverage to encourage the offender to participate in treatment while delivering punishment for wrongdoing and acting to protect the community. In the state of Victoria the CORE Sex Offender Programs have developed a statewide strategy in the public correctional system in order to assess, manage, and treat male sexual offenders. However, such offenders are notoriously reluctant to engage in treatment to address offending behaviors. A critical element of the strategy has been the Victorian Adult Parole Board, an entity that can determine that an offender needs to engage in treatment before he is considered for parole. Using the therapeutic jurisprudence framework as outlined by Wexler (1990), strategies to minimize the anti-therapeutic effects of the Victorian Adult Parole Board and maximize the therapeutic effects of the CORE Sex Offender Programs are highlighted. PMID- 11018781 TI - From therapeutic jurisprudence...to jurisprudent therapy. AB - For nearly a decade, "therapeutic jurisprudence" (TJ) has provided a theoretical framework within which legal rules, legal procedure, and legal roles are analyzed in terms of their therapeutic, neutral, or antitherapeutic effects. This article proposed "jurisprudent therapy" (JT), an extension of the TJ model, as a context for analyzing mental health science, mental health practice, and mental health roles in terms of their "jurisprudent," neutral, or "antijurisprudent" effects. The JT perspective neither supplants nor supersedes TJ; rather, it mirrors and augments an established process for interdisciplinary contrast, comparison, and integration. Just as an empathic and evolving legal system provides psychological benefits, so does a legally informed and juridically compatible progression of social science promote principles of justice and human freedom. Consideration of these two complementary models in tandem yields an array of brainstorming devices, to synergistic effect, with heuristic implications for teaching, research, and service delivery. PMID- 11018782 TI - Rethinking deprivations of liberty: possible contributions from therapeutic and ecological jurisprudence. AB - In place of the police and parens patriae powers, this article proposes three distinct justifactory models for government-sponsored deprivations of liberty. The punishment model authorizes deprivation of liberty as a sanction for blameworthy behavior. The prevention model authorizes deprivation of liberty to prevent harm, either through deterrence or restraint. The protection model authorizes liberty deprivation to ensure autonomous decisionmaking. The article compares these models to the purposes traditionally advanced as justification for punishment, and explores their strengths and weaknesses. Using therapeutic jurisprudence and ecological jurisprudence as organizing frameworks, it then describes a range of empirical issues raised by each of the models. PMID- 11018783 TI - The family group conference in the New Zealand children, young persons, and their families act of 1989 (CYP&F): review and evaluation. AB - The family group conference is an innovation introduced into New Zealand law as a means of resolving child protection and youth justice cases. The law requires the conference to include the offender, the victim, the extended family, and other relevant parties. This article reviews New Zealand research and commentary, and publications reporting on experiences with the methods in other countries. The discussion emphasizes problems in adapting a method based on one culture to a different culture and social organization. The article also considers the law from the viewpoint of therapeutic jurisprudence. Although the research is sparse, the FGC has strong promise for resolving problems, enhancing the sense of community and participation, and empowering families. PMID- 11018784 TI - In vivo PET studies of the dopamine D2 receptors in rhesus monkeys with long-term MPTP-induced parkinsonism. AB - Studies of dopamine (DA) receptor binding in early parkinsonian patients, or in models of Parkinson's disease, have revealed a supersensitivity of the D2-like receptor subtype as compared to age-matched controls. The lack of upregulation in advanced patients is often attributed to the effects of prolonged antiparkinsonian therapy, but the impact of therapy vs. intrinsic mechanisms in untreated patients or animals with long-term lesions of the DA nigrostriatal pathway has been difficult to address. We studied, in vivo, by PET using the DA D2 receptor ligand raclopride, the status of the DA receptors in normal rhesus monkeys and those with acute (3 months) or long-term (10 years) MPTP-induced nigrostriatal lesions. Compared to age-matched controls, there was no change in raclopride binding in MPTP-treated animals without parkinsonian symptoms. There was a significant increase in raclopride binding in the putamen (but not caudate nucleus) of all the animals displaying rigidity, hypo- and bradykinesia. This increase was greater in the animals with acute lesions (32%) than with established, long-term lesions (18%). There was no correlation between the postmortem striatal DA concentrations and in vivo raclopride binding but there was a correlation between PET raclopride binding and [(3)H]raclopride binding in vitro. Complex changes in D2 receptor binding occur in various stages of parkinsonism. Antiparkinsonian therapy is unlikely to be solely responsible for the lack of upregulation found in advanced parkinsonian patients but may be a contributing factor. PMID- 11018785 TI - GABA-containing neurons in the rat ventral tegmental area project to the prefrontal cortex. AB - Dopamine-containing projections from the ventral tegmental area (VTA) to the prefrontal cortex (PFC) have been extensively characterized since their discovery over 25 years ago. However, the VTA projection to the PFC also contains a substantial nondopamine component, whose neurochemical phenotype is unknown. To examine if a portion of this nondopamine VTA projection contains GABA, anterograde and retrograde tract-tracing in the rat was combined with GABA immunocytochemistry and electron microscopy. Following injections of Fluoro-Gold (FG) into the PFC, many VTA neurons were retrogradely labeled, as visualized by immunoperoxidase staining for FG. A large portion of FG-labeled somata (58%) and dendrites (33%) within the VTA also contained immunogold-silver labeling for GABA. These dually labeled profiles exhibited a morphology similar to dopamine containing cells within the VTA. To confirm and extend these findings, anterograde transport of biotinylated dextran amine (BDA) from the VTA was combined with immunogold-silver labeling for GABA within the PFC. Consistent with the results obtained from retrograde tracing, a portion of BDA-labeled terminals in the PFC also contained immunoreactivity for GABA. These dually labeled terminals formed symmetric synapses onto small caliber dendrites and dendritic spines. Some PFC dendrites contacted by GABA-containing VTA terminals were themselves GABA-labeled. The results of this investigation have identified a substantial population of GABA-containing neurons in the VTA that send axons to the PFC where they synapse on the distal processes of both pyramidal and local circuit neurons. This GABA-containing mesocortical pathway may provide substrates for both inhibitory and disinhibitory influences on PFC neuronal activity. PMID- 11018786 TI - Afferents from the auditory thalamus synapse on inhibitory interneurons in the lateral nucleus of the amygdala. AB - Physiological studies suggest that afferents to the lateral nucleus of the amygdala (LA) from the auditory thalamus initiate feedforward inhibition [Li et al. (1996b)]. This model of neural processing requires that thalamic afferents synapse directly onto inhibitory interneurons. To determine whether such synaptic contacts occur, we combined anterograde tract tracing with interneuron immunocytochemistry. The anterograde tracer biotinylated dextran amine (BDA) was injected into the auditory thalamus. Inhibitory interneurons in the LA were identified using antibodies directed against gamma aminobutyric acid (GABA) or one of the calcium binding proteins (CBPs), parvalbumin (PARV), calbindin (CALB), or calretinin (CALR), since CBPs identify distinct populations of GABAergic cells within the amygdala. The distribution of GABAergic and CBP interneurons in each subregion of the LA was examined by light microscopy and the relationships between thalamo-amygdala terminals and interneurons were examined by confocal and electron microscopy. Immunoreactive cells were distributed in all three subdivisions of LA, except for CALR-ir neurons, which were sparse in the dorsal subregion and were found mainly in the ventromedial and ventrolateral subregions. Confocal microscopy revealed some thalamo-amygdala terminals in close proximity to LA interneurons, while electron microscopy showed that thalamo-amygdala terminals made direct synaptic contacts onto distal dendritic processes of inhibitory neurons. These data provide morphological evidence that thalamic afferents synapse directly onto inhibitory interneurons in LA, and are consistent with the possibility that inputs from the auditory thalamus initiate feedforward inhibition in LA. This architecture could play an important role in the suppression of background neural noise, thereby enhancing the response of LA cells to incoming auditory stimuli. PMID- 11018787 TI - Effect of amphetamine on extrastriatal D2 dopamine receptor binding in the primate brain: a PET study. AB - [(11)C]raclopride binding to D2 dopamine receptors in the striatum is sensitive to drug-induced changes of endogenous dopamine concentration. We recently developed the new radioligand [(11)C]FLB 457, which is suitable for positron emission tomography (PET) studies of extrastriatal D2 dopamine receptors. The purpose of this PET study was to examine the effect of amphetamine on [(11)C]FLB 457 binding in extrastriatal regions. Each of three cynomolgus monkeys was examined at baseline conditions, 15 min and 3 h after I.V. injection of amphetamine (2 mg/kg). The effect of amphetamine was calculated from the ratio of specific [(11)C]FLB 457 binding to the binding in the cerebellum, a region which was used as reference for free and nonspecific binding in the brain. The changes of the ratio in the striatum, the thalamus, and the neocortex were between -1.2% and -15.5% at 15 min and -2.1% and -16.3% at 3 h, respectively, after amphetamine administration. The reductions of the binding ratios in the extrastriatal regions are similar to those reported for [(11)C]raclopride binding in the striatum. These data in a limited series of monkeys suggest that [(11)C]FLB 457 binding to D2 dopamine receptors in extrastriatal regions is sensitive to changes in the concentration of endogenous dopamine. PMID- 11018788 TI - Metyrapone, an inhibitor of corticosterone synthesis, blocks the kainic acid induced expression of HSP 70. AB - It is shown in the present study that metyrapone (100 mg/kg), an inhibitor of corticosterone synthesis, given twice, 30 min before and 6 h after kainic acid (10 mg/kg) administration, blocks the kainic acid-evoked induction of heat shock proteins 72 kDa (HSP 70). Specifically, it was observed that metyrapone completely prevented kainic acid-induced appearance of HSP 70 in the rat amygdala, habenula, parietal cortex, and significantly decreased the number of HSP 70-positive neurons in the CA1, CA3, and CA4 subregions of hippocampus. The reduction in HSP 70 induction was paralleled by a complete prevention of the kainic acid-induced rise in the circulating corticosterone level by metyrapone; however, in applied doses metyrapone evoked slight enhancement of blood corticosterone. Despite the fact that metyrapone blocked/attenuated the kainic acid-evoked induction of HSP 70, its administration did not affect the behavioral effects of kainic acid, regarded as "limbic status epilepticus." It is concluded that the blockade of corticosterone synthesis might have neuroprotective effects in the pathological states associated with the overstimulation of glutamatergic receptors. PMID- 11018789 TI - Muscarinic agonist carbachol depresses excitatory synaptic transmission in the rat basolateral amygdala in vitro. AB - Intracellular recordings in slice preparations of the basolateral amygdala were used to test which excitatory amino acid receptors mediate the excitatory postsynaptic potentials due to stimulation of the external capsule. These recordings were also used to examine the action of muscarinic agonists on the evoked excitatory potentials. Intracellular recordings from amygdaloid pyramidal neurons revealed that carbachol (2-20 microM) suppressed, in a dose-dependent manner, excitatory postsynaptic responses evoked by stimulation of the external capsule (EC). This effect was blocked by atropine. The estimated effective concentration to produce half-maximal response (EC(50)) was 6.2 microM. Synaptic suppression was observed with no changes in the input resistance of the recorded cells, suggesting a presynaptic mechanism. In addition, the results obtained using the paired-pulse protocol provided additional support for a presynaptic action of carbachol. To identify which subtype of cholinergic receptors were involved in the suppression of the EPSP, four partially selective muscarinic receptor antagonists were used at different concentrations: pirenzepine, a compound with a similar high affinity for muscarinic M1 and M4 receptors; gallamine, a noncompetitive antagonist for M2; methoctramine, an antagonist for M2 and M4; and 4-diphenylacetoxy-N-methylpiperidine, a compound with similar high affinity for muscarinic receptors M1 and M3. None of them independently antagonized the suppressive effect of carbachol on the evoked EPSP completely, suggesting that more than one muscarinic receptor subtype is involved in the effect. These experiments provide evidence that in the amygdala muscarinic agonists block the excitatory synaptic response, mediated by glutamic acid, by acting on several types of presynaptic receptors. PMID- 11018790 TI - Modulation of behavioral sensitization to cocaine by NAALADase inhibition. AB - Sensitization to cocaine has been attributed to alterations in excitatory amino acid and dopamine neurotransmission in the mesolimbic system. The present study sought to determine whether inhibition of NAALADase, an enzyme that cleaves glutamate from the endogenous neuropeptide, N-acetyl-aspartyl-glutamate (NAAG), attenuates sensitization to the psychomotor stimulant effects of cocaine. Rats received daily injections of cocaine (20.0 mg/kg/day; i.p.) or saline for 5 days. Fifteen minutes prior to these injections they received an i.p. injection of the NAALADase inhibitor, 2-PMPA (50.0-100 mg/kg), or vehicle. Locomotor activity and stereotypy produced by a challenge dose of cocaine (15.0 mg/kg) were assessed 3 days later. Acute cocaine administration increased locomotor activity in control animals. In animals with a prior history of cocaine administration, the behavioral response to cocaine was significantly enhanced. In animals that had received 2-PMPA in combination with cocaine, the enhancement of cocaine-induced locomotor activity was attenuated. No alteration in cocaine-evoked activity was observed in animals that had received once daily injections of 2-PMPA, alone. Acute administration of 2-PMPA also did not modify saline-induced locomotor activity or activity produced by an acute cocaine challenge. These data demonstrate that NAALADase inhibition attenuates the development of sensitization to the locomotor-activating effects of cocaine. Furthermore, this action cannot be attributed to an antagonism of the acute effects of cocaine. PMID- 11018791 TI - Comparison of the effects of sibutramine and other weight-modifying drugs on extracellular dopamine in the nucleus accumbens of freely moving rats. AB - The acute effects of systemic administration of the anti-obesity agent sibutramine on extracellular dopamine (DA) in the nucleus accumbens of freely moving rats were studied using in vivo microdialysis and compared with the actions of phentermine and d-amphetamine at doses 1x and 3x their respective 2 h ED(50) values to reduce food intake in rats. At the lower dose, sibutramine did not elevate extracellular DA concentrations; however, at the higher dose (6.0 mg kg(-1), i.p.) it caused a modest and prolonged increase in extraneuronal DA. A maximal rise was observed at 60 min post-sibutramine treatment (+231% compared to controls) with DA levels remaining elevated for up to 160 min post treatment. In contrast, phentermine and d-amphetamine significantly enhanced DA efflux at both the lower and higher doses. These elevations of DA levels were significantly greater than that seen with the corresponding dose of sibutramine over 0-80 min post treatment. Maximal rises in DA levels resulting from the higher dose of each drug were +733% (phentermine, 3.9 mg kg(-1), i.p.) and +603% (d-amphetamine, 1.5 mg kg(-1), i.p.) compared to controls 40 min post treatment. The highest doses of phentermine and d-amphetamine increased rat locomotor activity up to 100 min and 160 min post treatment, respectively, whereas the equivalent sibutramine dose had no effect. These findings therefore suggest that dopaminergic reward mechanisms are not involved in the reduction of food intake by sibutramine. Furthermore, they are consistent with the view that sibutramine lacks abuse potential. PMID- 11018792 TI - Receptor-mediated internalization of somatostatin in rat cortical and hippocampal neurons. AB - Binding of neuropeptides to their receptors usually results in internalization of receptor-ligand complexes. This process serves a crucial role in receptor downregulation, resensitization, and transmembrane signaling. It has mainly been investigated in cells ectopically expressing recombinant receptors. In the present study, we investigated whether rat central neurons and astrocytes naturally expressing somatostatin (SRIF) receptors internalized this neuropeptide. We demonstrated that 29% of cortical and 45% of hippocampal neurons in culture expressed the SRIF receptor sst(2A) and that 40-50% of the neurons internalized fluorescent SRIF. Similarly, an important proportion of astrocytes expressed sst(2A) (up to 60% in cortical cultures) and internalized fluo-SRIF. Competition experiments using the sst(2)/sst(5)-preferring agonist SMS 201-995 (octreotide) showed that a subpopulation of neurons internalized fluo-SRIF via sst(2) and/or sst(5) receptors, but that others also did so via other subtypes. Fluo-SRIF labeling was barely competed for by the sst(1)-selective agonist CH 275, indicating that sst(1) was unlikely to be mediating SRIF internalization in hippocampal and cortical neurons. Given the paucity of sst(5) receptors in cerebral cortex and hippocampus and the poor yield of sst(4) internalization in transfected cells, we conclude that sst(2) and sst(3) subtypes are the most likely to be responsible for SRIF internalization in our culture systems. PMID- 11018793 TI - Endogenous Zn(2+) is required for the induction of long-term potentiation at rat hippocampal mossy fiber-CA3 synapses. AB - The functional role of the abundant Zn(2+) found in some hippocampal synapses has been an enigma. We show here, using N-[6-methoxy-8-quinolyl]-P-toluenesulfonamide (TSQ) staining, that chelatable-Zn(2+) can be removed from hippocampal synaptic boutons using dietary depletion or with Zn(2+) chelators. A chronic dietary deficiency of bouton Zn(2+) resulted in the impairment of long-term potentiation (LTP) at mossy fiber-CA3 synapses. The averaged normalized fEPSP slope 30 min after tetanus was 209 +/- 28% of baseline value in control (mean +/- SEM, n = 10), and 118 +/- 12% in Zn(2+)-deficient rats (mean +/- SEM, n = 12, P < 0.01). In the deficient rats with Zn(2+) supplements, mossy fiber LTP returned to normal levels. The acute depletion of bouton Zn(2+) in the hippocampal slice with membrane-permeable Zn(2+) chelators, dithizone, or diethyldithiocarbamic acid (DEDTC) blocked the induction of mossy fiber LTP. The mean amplitudes of EPSCs after tetanus were 194 +/- 22% of baseline value in control (n = 5), compared to 108 +/- 14% in dithizone (n = 6) and 101 +/- 12% in DEDTC (n = 5). The averaged value of LTP, at the associational commisural fiber-CA3 synapses, was 193 +/- 20% in the control (n = 6), compared to 182 +/- 21% (n = 6, P > 0.1) in the presence of dithizone. The blockade of mossy fiber LTP by dithizone was reversible after washout. In addition, normal LTP could be induced by tetanus if exogenous Zn(2+) was applied immediately following dithizone. Our results indicate that the endogenous Zn(2+) is specifically required for LTP induction at the mossy fiber input into CA3 neurons. PMID- 11018794 TI - Effects of cocaine administration on receptor binding and subunits mRNA of GABA(A)-benzodiazepine receptor complexes. AB - The effects of intermittent intraperitoneal (i.p.) administration of cocaine (20 mg/kg) on GABA(A)-benzodiazepine (BZD) receptors labeled by t [(35)S]butylbicyclophosphorothionate (TBPS), and on several types of mRNA subunits were investigated in rat brain by in vitro quantitative receptor autoradiography and in situ hybridization. Phosphor screen imaging with high sensitivity and a wide linear range of response was utilized for imaging analysis. There was a significant decrease in the level of alpha 1, alpha 6, beta 2, beta 3, and gamma 2 subunits mRNA, with no alteration of [(35)S]TBPS binding in any regions in the brain of rats at 1 h following a single injection of cocaine. In chronically treated animals, the mean scores of stereotyped behavior were increased with the number of injections. The level of beta 3 subunit mRNA was decreased in the cortices and caudate putamen, at 24 h after a final injection of chronic administrations for 14 days. In the withdrawal from cocaine, the frontal cortex and hippocampal complexes showed a significant increase in [(35)S]TBPS binding and alpha1 and beta 3 subunit mRNA in the rats 1 week after a cessation of chronic administration of cocaine. These findings suggest that the disruption of GABA(A)-BZD receptor formation is closely involved in the development of cocaine-related behavioral disturbances. Further studies on the physiological functions on GABA(A)-BZD receptor complex will be necessary for an explanation of the precise mechanisms underlying the acute effects, development of hypersensitization, and withdrawal state of cocaine. PMID- 11018795 TI - Serotonin 2A and 2C receptor biosynthesis in the rodent striatum during postnatal development: mRNA expression and functional linkage to neuropeptide gene regulation. AB - The present study was designed to determine if there are region-specific differences in serotonin (5-HT) neurotransmission and 5-HT receptor expression that may limit the stimulatory effects of the 5-HT releaser p-chloroamphetamine (pCA) on striatal neuropeptide gene expression to the posterior striatum (P-STR) during postnatal maturation. Sprague-Dawley rat brains from postnatal days (PND) 1-35 were processed for 5-HT(2A) and 5-HT(2C) receptor mRNA expression by in situ hybridization and monoamine analysis by HPLC. Within the P-STR, 5-HT(2A) receptor mRNA expression reached young adult (PND 35) levels by PND 3, while levels in the A-STR were significantly less (range: 1.43 +/- 0.219-6. 36 +/- 0.478) than P-STR (5.36 +/- 0.854-12.11 +/- 1.08) at each respective age throughout the time course. 5-HT(2C) receptor mRNA expression reached young adult levels at PND 7 in the A-STR and by PND 3 in the P-STR. At each PND age 5-HT(2C) receptor mRNA levels within the P-STR were significantly less (6.23 +/- 1.02-12.32 +/- 0.427) than the A-STR (7.31 +/- 1.65-26.84 +/- 2.24). 5-HT content increased across the developmental time course within the P-STR (5.01 +/- 0.327-15.7 +/- 1.03 ng/mg protein) and A-STR (2.97 +/- 0. 223-11.2 +/- 0.701 ng/mg protein). Four hours following injection (i. p.) of pCA (10 mg/kg), preprotachykinin (PPT) mRNA levels increased 89% in the P-STR but not the anterior (A-STR) striatum of the 3-week old rat, which were prevented by preinjection (30 min, i.p.) of the 5-HT(2) receptor antagonist ritanserin (1 mg/kg). Together, these data suggest that faster maturity of 5-HT(2A) receptor expression in the P-STR may be sufficient to convey the region-specific acute stimulatory effects of pCA on PPT mRNA transcription in the developing rodent striatum. These results provide further evidence that the influence of 5-HT on neuropeptide gene expression is far stronger in caudal vs. rostral striatal regions during postnatal development. PMID- 11018796 TI - Opiate receptor avidity in the thalamus is sexually dimorphic in the elderly. AB - Opiate receptor avidity (B(')(max)/K(D)), was measured in the subcortex of nine females (five healthy subjects, four Alzheimer patients) and 15 males (seven healthy subjects, eight Alzheimer patients), 51-75 years of age, with the opiate receptor antagonist 6-deoxy-6-beta-[(18)F]fluoronaltrexone (cyclofoxy, CF) and a positron emission tomograph. CF avidity was 27.5% less in the thalamus of healthy women compared to healthy men and 48.5% less in Alzheimer disease female patients compared to male patients. PMID- 11018797 TI - IgM antiavian antibodies in sera from patients with pigeon breeder's disease. AB - The authors' objective was to study the presence of IgM antiavian antibodies in sera from patients with pigeon breeder's disease. We studied 93 patients with interstitial lung disease admitted for the assessment of pigeon breeder's disease. Eighty sera from healthy donors with no history of bird contact and 47 asymptomatic pigeon breeders were included as controls. The presence of IgM, IgG, and IgA antiavian antibodies was detected by ELISA and Western blot using avian pooled serum antigen. Fifty-three patients were classified as having definite pigeon breeder's disease, whereas 40 did not fulfill these diagnostic criteria. The levels of IgM antiavian-antibodies in pigeon breeder's disease by ELISA exceeded both the values of healthy subjects with no history of avian contact (P = 2.5 x 10(-8)) and the results of asymptomatic breeders (P = 0. 03). Positive IgA antiavian antibodies were the most frequent abnormalities in pigeon breeder's disease showing values over the reference levels of control groups that reach significant statistical differences. Both precipitin-positive and -negative samples demonstrated IgM reactivity. IgM antiavian antibodies were confirmed by Western blot. A relationship of IgM positive tests with a recent history of avian antigen exposure and acute disease was found. Additionally, the positive IgM group included patients having subacute and chronic lung disease. Antiavian antibodies have previously been considered of minor significance in hypersensitivity pneumonitis; nevertheless, recent studies support their use in clinical diagnosis. Although no specific laboratory tests can confirm the diagnosis in pigeon breeder's disease, IgM antiavian antibodies may be useful for detecting recent antigen exposure and the acute stage of the disease. PMID- 11018798 TI - Serial changes in enzyme inhibitory antibody to pyruvate dehydrogenase complex during the course of primary biliary cirrhosis. AB - To assess the usefulness of enzyme inhibition assay for the diagnosis of primary biliary cirrhosis (PBC), we determined the serial changes in enzymatic inhibitory antibody to pyruvate dehydrogenase complex (PDC) in patients with PBC, and compared the results to those of immunofluorescence and immunoblotting. Forty nine sera from 19 patients with PBC who were followed-up for at least 16 months were tested for antimitochondrial antibodies (AMA) by indirect immunofluorescence, immunoblotting on bovine heart mitochondria, and enzyme inhibition assay using commercially available TRACE Enzymatic Mitochondrial Antibody (M2) Assay (EMA) kit. Of the 49 sera, 39 (80%), 35 (71%), 38 (78%), 31 (63%), and 36 (73%) were positive for AMA by immunofluorescence, for immunoglobulin G (IgG), IgM, and IgA class antibody against E2 subunit of PDC (PDC-E2) by immunoblotting, and for enzymatic inhibitory antibody to PDC by EMA, respectively. AMA titers determined by immunofluorescence did not change in 9 patients (47%), increased in 4 (21%), decreased in 3 (16%), and fluctuated in 3 (16%) during follow-up. The number of anti-M2 bands by immunoblotting did not change in 9 (47%), increased in 6 (32%), decreased in 2 (11%), and fluctuated in 2 (11%). Units of PDC activity by EMA did not change markedly in 16 (84%), increased in 2 (11%), and fluctuated in 1 (5%). Positive EMA results were common in cases with high levels of serum alkaline phosphatase and IgM, and the units of PDC activity by EMA correlated significantly and inversely with AMA titers by immunofluorescence, and serum reactivity to PDC-E2 by immunoblotting, respectively. There was no correlation between serial changes in biochemical data and units of PDC activity by EMA. In three patients who showed a decrease in AMA titers, AMA titers correlated more with EMA results than immunoblotting. Moreover, in a patient with fluctuating AMA titers, the units of PDC activity by EMA paralleled AMA titers. Our results suggest that EMA is useful for the diagnosis of AMA-positive PBC, and also could be used for monitoring the disease course in PBC. PMID- 11018799 TI - Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction. AB - The development of a multiplex polymerase chain reaction method for the rapid and accurate detection and typing of HSV-1, HSV-2, and VZV from clinical specimens is described. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations. False-negative results that sometimes arise due to inhibitors of DNA amplification or failure of DNA extraction procedure used may be avoided by assaying each specimen with alpha-tubulin primers. Multiplex PCR amplified viral sequences from all 55 specimens obtained from patients with clinical evidence of HSV or VZV infection indicated 100% sensitivity. From 55 patients who were investigated by multiplex PCR, HSV-1 was detected in 28, HSV-2 in 20, and VZV in 7 specimens. The reported results indicate that the present multiplex PCR assay has a potential application in clinical diagnosis when a rapid and accurate detection and typing of involved viruses HSV-1, HSV-2, or VZV is needed. PMID- 11018800 TI - Measurement of serum IgA and C3 may predict the diagnosis of patients with IgA nephropathy prior to renal biopsy. AB - The levels of serum IgA and C3 in patients with IgA nephropathy were determined using international standard serum (IFCC/CRM470) in a multicenter trial in Japan. The ratio of serum IgA to C3 (serum IgA/C3 ratio) without any information from renal biopsy was used for the diagnosis of IgA nephropathy. Three hundred and six patients with IgA nephropathy and other glomerular diseases, and 418 healthy adults were examined. The new diagnostic standardized criterion in patients with IgA nephropathy, obtained by nephelometric immune assay based on the international reference preparation CRM470, was 315 mg/dl. The serum IgA/C3 ratio was a more useful marker for distinguishing IgA nephropathy from non-IgA nephropathy together with serum IgA levels. This suggests that the measurement of serum IgA and C3 may predict the diagnosis of patients with IgA nephropathy prior to renal biopsy. PMID- 11018801 TI - Performance evaluation of a new chemiluminescent cardiac troponin I assay. AB - Cardiac troponin I is a marker for diagnosis of myocardial damage. Several immunoassays are currently available for determination of concentrations of troponin I in serum. We evaluated a chemiluminescent assay for troponin I using ACS:180 automated analyzer (Bayer Diagnostics). We compared our results with two other immunoassays using the OPUS Magnum (OPUS troponin I assay, Dade Behring) and AxSYM (microparticle enzyme immunoassay, Abbott) analyzers. The within-run and between-run CVs were less than 5% for all three levels of controls. The chemiluminescent assay for troponin I was linear up to a serum troponin I concentration of 50 ng/mL and the detection limit was 0.1 ng/mL of troponin. A good correlation between troponin I concentration measured by the chemiluminescent assay (y axis) and the microparticle enzyme immunoassay (MEIA) (x axis) was observed, although the concentrations of troponin I in individual specimens were approximately four times higher, when measured by the MEIA assay, than those measured by chemiluminescent assay. The correlation coefficient was 0.98 with the regression equation y = 0.22x + 1.125. We also observed a good correlation in troponin I concentrations obtained by the chemiluminescent assay (y axis) and OPUS troponin I assay (x axis). The correlation coefficient was 0.96 and the regression equation was y = 0.79x - 0.52. The correlation coefficient was 0.93 when we compared troponin I concentrations obtained by the OPUS assay (x axis) with the corresponding concentrations obtained by the MEIA assay (y axis). The corresponding regression equation was y = 0.25x + 3.5. We conclude that the chemiluminescent troponin I assay showed good analytical performance. PMID- 11018802 TI - ACE and AT1R gene polymorphisms and hypertension in Indian population. AB - The renin angiotensin system (RAS) controls intrarenal blood pressure and sodium balance, and is an important target for antihypertensive therapy. Several polymorphisms have been identified within genes encoding RAS that may contribute to the development of elevated blood pressure. The relevance of these polymorphisms in hypertension remains controversial. In this study we have examined 105 hypertensive subjects and 192 controls from the Indian population for I/D polymorphism of angiotensin I converting enzyme (ACE) and A(1166)C polymorphism of angiotensin II type I receptor (AT1R) genes by polymerase chain reaction (PCR) and PCR-based restriction enzyme analysis method, respectively. There was no significant difference in the distribution of ACE (I/I, I/D, and D/D) and AT1R (A/A and A/C) genotypes between controls and hypertensive subjects. D allele was significantly associated with an early onset of hypertension and although nonsignificant, the frequency was high in subjects with family history of cardiovascular disorders. C(1166) allele of AT1R did not correlate with the age of onset of hypertension and the frequency was low in subjects with family history. Thus no association was found between ACE and AT1R genotypes and hypertension. However the D allele can be used as a predictor of risk of hypertension in the Indian population. PMID- 11018803 TI - Evaluation of Henes-PCR assay for Mycobacterium detection in different clinical specimens from patients with or without tuberculosis-associated HIV infection. AB - The need for early diagnosis of tuberculosis, particularly in HIV-infected patients, requires the development of diagnosis methods that have a high sensitivity and specificity, as does the nucleic acid-based technology. With the purpose of improving the detection of mycobacterium in different clinical samples, we proposed and evaluated an assay based on nucleic acid-amplification: heminested-PCR (Henes-PCR). The procedure was designed to identify Mycobacterium spp., M. tuberculosis complex (MTC), and M. avium complex (MAC), although it has the potential to include more primers for the identification of other species. Analytical and clinical evaluation of Henes-PCR was performed by analysis of reference strains and 356 clinical specimens from 246 patients with pulmonary and meningitis tuberculosis and unrelated infections, including 142 HIV-infected individuals. Ninety-three percent (199) positive and 100% (143) negative results were obtained in specimens from patients with tuberculosis and non-tuberculosis infection, respectively. The overall sensitivity of Henes-PCR was 93.4%, specificity was 100%, positive and negative predictive values were 100 and 91.1%, respectively. Sensitivity and negative predictive value of Henes-PCR were significantly higher than culture procedure for microscopy-negative specimens. Even though frequency of HIV infection was higher in patients with tuberculosis, diagnostic parameters of Henes-PCR were similar between HIV-positive and HIV negative patients. MTB was identified in 194 (98%) specimens while MAC was detected in 5 (2%) specimens. These findings suggest that Henes-PCR is a useful test for rapid detection of mycobacterium in clinically suspected cases of tuberculosis with smear-negative results. PMID- 11018804 TI - Higher prevalence of urogenital mycoplasmas in human immunodeficiency virus positive patients as compared to patients with other sexually transmitted diseases. AB - We determined the prevalence of three emergent urogenital mycoplasma species (M. fermentans, M. penetrans, and M. genitalium) in comparison to the most common species (M. hominis and U. urealyticum). M. genitalium is probably the third most frequent agent of nongonococcal urethritis (NGU) in men. It has been suggested that M. fermentans and M. penetrans play a role in the development of AIDS. We analysed the urine and the urethral swab samples from 106 HIV-1 infected individuals (HIV group) and 110 HIV-negative patients with NGU (STD group) by using PCR and culture methods. M. genitalium was detected in 0.9% of the urine and in 1.9% of the urethral samples from the HIV group, compared to 9.1% found only in urethral swab samples from the STD group. M. fermentans was detected in 5.7% urethral swabs from the HIV group and in 0.9% from the STD group. M. penetrans was detected in 6.6% urine samples from the HIV group. M. hominis and U. urealyticum showed infection rates of 7.5 and 18.9% in the HIV group, and 0.9 and 13.6% in the STD group. Overall there was a higher prevalence of mycoplasmas in the HIV group than in the STD group, but the significance of these results remains unclear. PMID- 11018805 TI - Ambulatory blood pressure monitoring and microalbuminuria in normotensive subjects with insulin-dependent diabetes mellitus. AB - OBJECTIVE: To assess the association between microalbuminuria with ambulatory blood pressure monitoring in normotensive individuals with insulin-dependent diabetes mellitus. METHODS: Thirty-seven patients underwent determination of the rate of urinary excretion of albumin through radioimmunoassay and ambulatory blood pressure monitoring. Their mean age was 26.5+/-6.7 years, and the mean duration of their disease was 8 (1-34) years. Microalbuminuria was defined as urinary excretion of albumin > or =20 and <200 microg/min in at least 2 out of 3 urine samples. RESULTS: Nine (24.3%) patients were microalbuminuric. Ambulatory blood pressure monitoring in the microalbuminuric patients had higher mean pressure values, mainly the systolic pressure, during sleep as compared with that in the normoalbuminuric patients (120.1+/-8.3 vs 110.8+/-7.1 mmHg; p=0.007). The pressure load was higher in the microalbuminuric individuals, mainly the systolic pressure load during wakefulness [6.3 (2.9-45.9) vs. 1.6 (0-16%); p=0.001]. This was the variable that better correlated with the urinary excretion of albumin (r(S)=0.61; p<0.001). Systolic pressure load >50% and diastolic pressure load > 30% during sleep was associated with microalbuminuria (p=0.008). The pressure drop during sleep did not differ between the groups. CONCLUSION: Microalbuminuric normotensive insulin-dependent diabetic patients show greater mean pressure value and pressure load during ambulatory blood pressure monitoring, and these variables correlate with urinary excretion of albumin. PMID- 11018806 TI - Reduction of QTc interval dispersion. Potential mechanism of cardiac protection of pyridostigmine bromide. AB - OBJECTIVE: Parasympathetic dysfunction is an independent risk factor in individuals with coronary artery disease, and cholinergic stimulation is a potential therapeutical option. We determined the effects of pyridostigmine bromide, a reversible anticholinesterase agent, on electrocardiographic variables of healthy individuals. METHODS: We carried out a cross-sectional, double blind, randomized, placebo-controlled study. We obtained electrocardiographic tracings in 12 simultaneous leads of 10 healthy young individuals at rest before and after oral administration of 45 mg of pyridostigmine or placebo. RESULTS: Pyridostigmine increased RR intervals (before: 886+/-27 ms vs. after: 1054+/-37 ms) and decreased QTc dispersion (before: 72+/-9 ms vs. after: 45+/-3 ms), without changing other electrocardiographic variables (PR segment, QT interval, QTc, and QT dispersion). CONCLUSION: Bradycardia and the reduction in QTc dispersion induced by pyridostigmine may effectively represent a protective mechanism if these results can be reproduced in individuals with cardiovascular diseases. PMID- 11018807 TI - Prognostic factors of rheumatic mitral stenosis during pregnancy and puerperium. AB - OBJECTIVE: To identify characteristics associated with complications during pregnancy and puerperium in patients with rheumatic mitral stenosis. METHODS: Forty-one pregnant women (forty-five pregnancies) with mitral stenosis, followed up from 1991 to 1999 were retrospectively evaluated. PREDICTOR VARIABLES: the mitral valve area (MVA), measured by echocardiogram, and functional class (FC) before pregnancy (NYHA criteria). Maternal events: progression of heart failure, need for cardiac surgery or balloon mitral valvulotomy, death, and thromboembolism. Fetal/neonatal events: abortion, fetal or neonatal death, prematurity or low birth weight (<2,500 g), and extended stay in the nursery or hospitalization in newborn ICU. RESULTS: The mean +/- SD of age of the patients was 28.8+/-4.6 years. The eventful and uneventful patients were similar in age and percentage of first pregnancies. As compared with the level 1 MVA, the relative risk (RR) of maternal events was 5.5 (95% confidence interval (CI) =0.8 39.7) for level 2 MVA and 11.4 (95% CI=1.7-74.5) for level 3 MVA. The prepregnancy FC (FC > or = II and III versus I) was also associated with risk for maternal events (RR=2.7; 95% CI=1.4-5.3).MVA and FC were not importantly associated with these events, although a smaller frequency of fetal/neonatal events was observed in patients who had undergone balloon valvulotomy. CONCLUSION: In pregnant women with mitral stenosis, the MVA and the FC are strongly associated with maternal complications but are not associated with fetal/neonatal events. Balloon mitral valvulotomy could have contributed to reducing the risks of fetal/neonatal events in the more symptomatic patients who had to undergo this procedure during pregnancy. PMID- 11018808 TI - The Bussola study. Final results, conclusions and proposals. AB - OBJECTIVE: To evaluate the use of thrombolytic and acetylsalicylic acid therapies in acute myocardial infarct patients as well as the availability of technical and human resources for the care of these patients in the emergency units of the city of Rio de Janeiro. Additional objectives were the evaluation of the use of primary angioplasty and the level of acceptance of SBC /RJ as an entity responsible for programs of continued medical education. METHODS: Interviews with physicians at 46 emergency units in the city of Rio de Janeiro. RESULTS: Of the 46 emergency units inspected, a policy of encouragement to use thrombolytic therapy was only prescribed in 6.5%. In 1/3 of the public wards no thrombolytic agents were available, and in none of them was access to primary angioplasty regularly available; 45.9% did not offer the minimal conditions required for the handling of cases of acute myocardial infarction; 60% of the physicians on-call (at both public and private emergency units), appeared not to know the importance of the use of acetylsalicylic acid in acute myocardial infarct patients; all physicians interviewed would participate in programs of continued medical education organized by the SBC/RJ. CONCLUSION: The study suggests there was: 1) the low probability of the use of thrombolytic therapy in the majority of the emergency units in of the city of Rio de Janeiro due to the inadequate policy of waiting for the transfer of the patient to coronary or intensive care unit; 2) a low awareness to the importance of early use of acetylsalic acid in acute myocardial infarct; 3) half of the emergency units of the public net do not have the minimal conditions required for the handling of cases of acute myocardial infarction; 4) a high level of credibility exists that would enable the SBC/RJ to set up programs for continued medical education to change the mentality regarding the use of thrombolytic therapy and of acetylsalicylic acid. PMID- 11018809 TI - Hormonal and cardiovascular reflex assessment in a female patient with pure autonomic failure. AB - We report the case of a 72-year-old female with pure autonomic failure, a rare entity, whose diagnosis of autonomic dysfunction was determined with a series of complementary tests. For approximately 2 years, the patient has been experiencing dizziness and a tendency to fall, a significant weight loss, generalized weakness, dysphagia, intestinal constipation, blurred vision, dry mouth, and changes in her voice. She underwent clinical assessment and laboratory tests (biochemical tests, chest X-ray, digestive endoscopy, colonoscopy, chest computed tomography, abdomen and pelvis computed tomography, abdominal ultrasound, and ambulatory blood pressure monitoring). Measurements of catecholamine and plasmatic renin activity were performed at rest and after physical exercise. Finally the patient underwent physiological and pharmacological autonomic tests that better diagnosed dysautonomia. PMID- 11018810 TI - Cardiac involvement in total generalized lipodystrophy (Berardinelli- Seip syndrome). AB - Total generalized lipodystrophy (Berardinelli - Seip Syndrome) is a rare hereditary disease characterized by insulin-resistant diabetes mellitus and a small quantity of adipose tissue and is of unknown origin. Common cardiovascular alterations related to this syndrome are cardiac hypertrophy and arterial hypertension. This article reports a case of Berardinelli - Seip syndrome and reviews the literature with special emphasis on the cardiovascular manifestations of this syndrome. PMID- 11018811 TI - Heart transplant in a patient with complete absence of the pericardium. AB - We report the case of a heart transplant in which the recipient patient had a total congenital absence of the pericardium. Associated with this, we found a major disproportion between the size of the recipient's mediastinal cavity and the size of the donor's heart. To prevent twisting of the great arteries, we placed the graft on the left diaphragm muscle and beneath the left lung, which resulted in an uneventful early and late postoperative course. PMID- 11018812 TI - Why are calcium antagonists still being used in heart failure in the era of calcium sensitizers? PMID- 11018813 TI - Diagnosis of dermatomyositis and polymyositis: a study of 102 cases. AB - Patients with dermatomyositis (DM) or polymyositis (PM) were studied retrospectively. The patients were divided into four groups: definite PM 24, probable PM 19, definite DM 34 and mild-early DM 25 cases. PM patients complained more often proximal muscle weakness [p <0.01]. DM patients complained more arthralgia [p <0.05], dysphagia [p <0.03] and weight loss [p <0.04]. Five patients had a malignant neoplasm and 9 had other connective-tissue disease. DM presented higher ESR than PM [p <0.002]. PM presented more significant increase in creatine kinase (CK) [p <0.02] and in alanine aminotransferase (ALT) [p <0.001] levels. Electromyography showed myopathic pattern in 76%. Muscle biopsy was the definitive test. Perifascicular atrophy was more frequent in definite DM than in mild-early DM group [p <0.03]. CONCLUSION: A small association with connective-tissue diseases and neoplasms was found. DM and PM are clinically different. DM presents systemic involvement affecting the skin, developing more severe arthralgia, dysphagia and weight loss and presenting higher values of ESR. PM presents a restricted and more significant involvement of muscles generating more weakness complaints and higher levels of serum muscle enzymes. PMID- 11018814 TI - The diagnosis of leprosy among patients with symptoms of peripheral neuropathy without cutaneous lesions: a follow-up study. AB - Forty-four patients with neuritic leprosy were individually followed for periods ranging from 4 months to almost 4 years for the purpose of ascertaining the presence and/ or absence of leprosy. The neural symptoms presented were sensory impairment (41), parasthesia (28), nerve enlargement (22), nerve tenderness (20), paresia (20), amyotrophy (8). Leprosy was diagnosed in ten out of the total number of patients studied. Leprosy was confirmed by the appearance of reactional neuritis (4), reversal reaction (2), biopsy of the hypoesthesic area (3) and the appearance of non-reactional cutaneous lesion. We reported an experience in the diagnosis of neuritic leprosy and its most frequent clinical presentation with which clinicians have to be acquainted. We can also state that the clinical follow-up was an effective strategy for the diagnosis of the disease when diagnostic facilities are not available or have not confirmed the diagnosis. PMID- 11018815 TI - Benign monomelic amyotrophy: a study of twenty-one cases. AB - A consecutive series of 21 patients with single limb atrophy (monomelic amyotrophy) is reported. Sixteen had lower limb atrophy and five had upper limb involvement. The median age of the onset was 20 years. Characteristic features were sporadic occurrence, wasting confined to one limb, insidious onset with slow progression, stabilizing in 1 to 4 years, and absence of pyramidal signs. All the patients with upper limb involvement were male, however in our cases with lower limb amyotrophy there were no male preponderance. We observed wasting of the entire length of the lower limbs in six patients. There were nine cases with amyotrophy restricted to the leg and one with amyotrophy only in the thigh. In the upper limb in four cases the involvement was distal and in one patient the atrophy was proximal. The electromyographic features were suggestive of anterior horn disease not only in the affected limb but also, in some cases, in clinically uninvolved limb. Cervical or lumbar MRI was normal. MRI of the lower limb disclosed increased signal intensity in the gastrocnemius and soleus muscles in one patient suggesting denervation. PMID- 11018816 TI - Non-progressive juvenile spinal muscular atrophy of the distal upper limb (Hirayama's disease): a clinical variant of the benign monomelic amyotrophy. AB - Hirayama's disease (HD) is frequently found in Asia, and is rarely referred among westerners. It affects young people with higher incidence in males. It is a focal distal amyotrophy with unilateral or asymmetric bilateral involvement of C7, C8 and T1 innervated muscles. HD appears sporadically and has a benign evolution with clinical stabilization in around one year. We report four young male patients with clinical and electrophysiological alterations described in HD, which were followed-up during 5 years. Electromyographic findings were indicative of lower motor neuron involvement. We analyzed cervical MRI aiming at understanding if a questionable spinal cord compression could be implicated in the pathogenesis, but no abnormality was verified. In view of its clinical, and EMG characteristics, HD is no more than a benign monomelic amyotrophy (BMA) clinical variant, and not a specific disease. This eponym could be considered only for the distal upper limb variant (Hirayama's variant) of the BMA. PMID- 11018817 TI - Scanning electron microscopy study of the choroid plexus in the monkey (Cebus apella apella). AB - The cells of the choroid plexus of the lateral ventricles of the monkey Cebus apella apella were examined through scanning electron microscopy at contributing to the description of such structures in primates. The animals were anesthetized previously with 3% hypnol intraperitoneally and after perfusion with 2.5% glutaraldehyde, samples of the choroid plexus were collected after exhibition of the central portion and inferior horn of the lateral ventricles. The ventricular surface of those cells presents globose form as well as fine interlaced protrusions named microvilli. Among those, it is observed the presence of some cilia. Resting on the choroid epithelial cells there is a variable number of free cells, with fine prolongations which extend from them. They are probably macrophages and have been compared to Kolmer cells or epiplexus cells, located on choroid epithelium. The choroid plexus of the encephalic lateral ventricles of the monkey Cebus apella apella at scanning electron microscopy is similar to that of other primates, as well as to that of other species of mammals mainly cats and rats, and also humans. PMID- 11018818 TI - Trail making and cognitive set-shifting. AB - We tested the hypothesis that Part B of the Trail Making Test (TMT) is a measure of cognitive set-shifting ability in 55 normal subjects with the conventional (written) TMT and a verbal adaptation, the "verbal TMT" (vTMT). The finding of a significant association between Parts B of TMT and vTMT (r = 0,59, p < 0,001), after correcting for age and education, supports the view that Part B of TMT is a valid measure of the ability to alternate between cognitive categories. PMID- 11018819 TI - Motor performance after posteroventral pallidotomy and VIM-thalamotomy in Parkinson's disease: a 1-year follow-up study. AB - Twenty-three patients with Parkinson's disease underwent stereotactic surgery. To study the long-term motor performance, the patients were evaluated at the pre operative period and at the 1st, 3rd, 6th, and 12th post-operative months, with the following scales: Unified Parkinson's Disease Rating Scale (UPDRS) motor score and Larsen's Scale for Dyskinesias. The patients under levodopa therapy were assessed both in "on" and "off" periods. Fourteen unilateral ventrolateral thalamotomies (VLT), 4 unilateral posteroventral pallidotomies (PVP), 2 bilateral PVP, and 3 VLT with contralateral PVP were performed. The motor improvement was significant and long-lasting in the "off" period, except for 2 patients. The "on" period quality improved, mainly due to the control of dyskinesias. The improvement of dyskinesias was long-lasting for the majority of the patients. There was no significant decrease in the levodopa dose. Three patients showed permanent complications, but none was severe. PMID- 11018820 TI - Polymerase chain reaction for the laboratory diagnosis of aseptic meningitis and encephalitis. AB - A protocol for testing cerebrospinal fluid specimens using a range of PCR assays for the diagnosis of central nervous system infection was developed and used to test prospectively 383 specimens. PCR assays were used for the detection of adenovirus, Borrelia burgdorferi, enteroviruses, Epstein Barr virus, cytomegalovirus, herpes simplex virus, human herpes virus type 6, JC virus, Leptospira interrogans, Listeria monocytogenes, lymphocytic choriomeningitis virus, measles virus, mumps virus, Mycobacterium sp. , Mycoplasma pneumoniae, Toxoplasma gondii and varicella zoster virus. Of the 383 specimens tested in this study, 46 (12.0%) were found to be positive. The microorganisms detected were CMV, enterovirus, Epstein Barr virus, herpes simplex virus, human herpes virus type 6, JC virus, L. monocytogenes, Mycobacterium genus, Toxoplasma gondii and varicella zoster virus. The introduction of the PCR protocol described has improved the diagnosis of a range of central nervous system infections in our laboratory. We believe however that further evaluation of these assays in immunocompromised patients is necessary to better determine the predictive value of positive PCR results in these patient groups. PMID- 11018821 TI - Staphylococcus aureus meningitis in children: a review of 30 community-acquired cases. AB - In spite of the steady increase in the incidence of Staphylococcus aureus infections, it remains a relatively uncommon cause of meningitis. To our knowledge, no series of community-acquired S. aureus meningitis (CASAM) restricted to children has been published. So far in this retrospective study we report our experience with CASAM in children, hospitalized from 1983 to 1998 at Nossa Senhora da Gloria Children's Hospital (HINSG). During the sixteen-year study period, 2,319 new cases of acute pyogenic meningitis were diagnosed at HINSG. Community-acquired S. aureus was identified as the causative agent in 30 patients (1.3 percent). The predominantly spinal localization of the agent is stressed. In contrast with publications which analyze adults, it has a better prognosis. PMID- 11018822 TI - Benign rolandic epilepsy: clinical and electroencephalographic correlates. AB - Benign rolandic epilepsy (BRE) is known for its dissociation from structural alterations. Nevertheless, the number of cases with reported organic lesions has been increasing. This led to the creation of two subgroups, "benign" and "non benign" BRE, and resulted in the need for additional parameters to define electrographic benignity. We assessed the possible associations between interictal electroencephalographic findings and clinical behavior in 60 BRE cases, testing four parameters of electrographic benignity (paroxysm morphology, horizontal dipole, base rhythms, laterality of rolandic spikes). We also assessed the relationship between neuroimaging findings and electrographic and clinical classifications, and found a statistically significant association (sensitivity=73.5%; specificity=81.8%; positive predictive value=94. 8%; negative predictive value=40.9%). Three of the electrographic parameters proposed were associated with clinical classification: paroxysm morphology, horizontal dipole, and base rhythms. Cases electrographically classified as benign have 21 times more chances to be equally classified as clinically benign according with the tested criteria. PMID- 11018823 TI - [Familial partial epilepsies]. AB - OBJECTIVE: To investigate the clinical and genetic characteristics of familial partial epilepsies. METHOD: Family history of seizures was questioned in all patients followed in our epilepsy clinics, from October 1997 to December 1998. Those with positive family history were further investigated and detailed pedigrees were obtained. All possibly affected individuals available underwent clinical evaluation. Seizures and epilepsy syndromes were classified according to the ILAE recommendations. Whenever possible, EEG and MRI were performed. RESULTS: Positive family history was identified in 32 unrelated patients. A total of 213 possibly affected individuals were identified, 161 of whom have been evaluated. The number of affected subjects per family ranged from two to 23. Temporal lobe epilepsy (TLE) was identified in 22 families (68%), frontal lobe epilepsy in one family (3%), partial epilepsy with centrotemporal spikes in five families (15%), and other benign partial epilepsies of childhood in four families (12%). Most of the affected individuals in the TLE families (69%) had clinical and/or EEG characteristics of typical TLE. However, the severity of epilepsy was variable, with 76% of patients with spontaneous seizure remission or good control with medication and 24% with refractory seizures, including 7 patients that underwent surgical treatment. In the other 10 families, we identified 39 possibly affected subjects, 23 of whom were evaluated. All had good seizure control (with or without medication) except for one patient with frontal lobe epilepsy. Pedigree analysis suggested autosomal dominant inheritance with incomplete penetrance in all families. CONCLUSION: Family history of seizures is frequent among patients with partial epilepsies. The majority of our families had TLE and its expression was not different from that observed in sporadic cases. The identification of genes involved in partial epilepsies may be usefull in classification of syndromes, to stablish prognosis and optimal treatment. PMID- 11018824 TI - [Brain hypometabolism in patients with mesial-temporal sclerosis demonstrated by FDG-PET]. AB - The purpose of this study was to evaluate the extent of brain hypometabolism in patients with mesial-temporal sclerosis (MTS). METHOD: This retrospective study included 21 patients who had medically refractory complex partial seizures and were selected for surgical therapy after a comprehensive evaluation which included surface EEG recordings and neuroimaging studies (PET, SPECT and MRI). All patients were subjected to surgical intervention and had an histopathological confirmation of MTS. A semi-quantitative analysis of the PET images was performed using regions of interest in the following structures: frontal, parietal and occipital lobes, basal ganglia, thalami, cerebellum and three different regions in the temporal lobes, which included medial, inferior and lateral cortices. An asymmetry index (AI) was calculated, comparing the counts per pixel in the homologous structures in both brain hemispheres. The AI of the different structures were then correlated. RESULTS: A significant correlation was demonstrated between the AI of the medial temporal cortices and the frontal lobe, parietal lobe, basal ganglia and thalami (r = 0.72, 0.62, 0.47 and 0.47 respectively with p < 0.05 ). Within the temporal lobe, highly significant correlations were demonstrated among the structures (as high as 0.86 between temporo-medial and temporo-inferior). CONCLUSION: These data indicated that hypometabolism extends beyond the epileptogenic focus in the temporal lobe in patients with complex partial seizure related to MTS. The metabolism in the medial portion of the temporal lobe is more correlated with the metabolism in the frontal lobe than with those of the others brain structures outside the temporal lobe. The pathophysiological mechanisms of hypometabolism remain controversial. PMID- 11018825 TI - [Cerebral hemometabolism: variability in the acute phase of traumatic coma]. AB - OBJECTIVE: to evaluate the interrelationships between cerebral and systemic hemometabolic alterations in patients with severe traumatic brain injury managed according to a standardized therapeutic protocol. DESIGN: prospective, interventional study in patients with traumatic coma. SETTING: a general Intensive Care Unit in a teaching hospital. PATIENTS AND METHODS: twenty-seven patients (21M e 6F), aging 14 - 58 years, with severe acute brain trauma, presenting with three to eight points on the Glasgow Coma Scale, were prospectively evaluated according to a cumulative protocol for the management of acute intracranial hypertension, where intracranial pressure (ICP) and cerebral extraction of oxygen (CEO2) were routinely measured. Hemometabolic interrelationships involving mean arterial pressure (MAP), ICP, arterial carbon dioxide tension (PaCO2), CEO2, cerebral perfusion pressure (CPP) and systemic extraction of oxygen (SEO2) were analyzed. INTERVENTIONS: routine therapeutic procedures. RESULTS: no correlation was found between CEO2 and CPP (r = -0.07; p = 0.41). There was a significant negative correlation between PaCO2 and CEO2 (r = -0.24; p = 0.005) and a positive correlation between SEO2 and CEO2 (r = 0.24; p = 0.01). The mortality rate in this group of patients was 25.9% (7/27). CONCLUSION: 1) CPP and CEO2 are unrelated; 2) CEO2 and PaCO2 are closely related; 3) during optimized hyperventilation, CEO2 and SEO2 are coupled. PMID- 11018826 TI - [Neurocysticercosis: a clinical and pathological study of 27 necropsied cases]. AB - Neurocysticercosis is the most frequent and widespread neuroparasitosis of the human being. The development of brain and leptomeningeal lesions, with subsequent symptoms, are mainly related with the immune status of the host, and to the number and evolutional phase of the parasites. We present the pathological findings in 27 necropsies of patients with neurocysticercosis, which accounted for 3.1% of the necropsies. 77% of the patients were male and the age ranged from 18 to 85 years. In 26% there was previous history of alcoholism. Clinicopathological study showed that 50% of the cases were classified as asymptomatic form, 11% epileptic form, 11% intraventricular form and 11% combined form. 33% of the patients presented seizures as a factor of aggravation of the clinical picture. There was a single cysticercus in 60% of the cases, the cellulosae form present in 82% and the racemous form in 7% of the cases; the remaining 11% had both forms present. In 30% of the patients the cause of death was directly related with the presence of the cysticercus in the central nervous system. Our findings confirm the high morbidity of this disease. PMID- 11018827 TI - [Malformations of the central nervous system: analysis of 157 pediatric autopsies]. AB - The malformations of the central nervous system affect about 5 to 10 children per 1000 births. We studied the central nervous system malformations in 5837 pediatric autopsies performed in the Sector of Anatomic Pathology, Hospital de Clinicas-UFPR, between 1960 and 1995. There were 157 central nervous system malformations (2.69%), the commonest were neural tube defects (61%): 47 cases of anencephaly and 45 cases classified in the group of myeloencephaloceles. The anomalies of the prosencephalic evagination corresponded to 8% of all central nervous system malformations, with seven cases of holoprosencephaly. Posterior fossa malformations occurred in 3%, with three cases of Arnold-Chiari. In the present study, the mortality rate due to central nervous system malformations was higher in the neonatal period. PMID- 11018828 TI - Cerebellar vermis hypoplasia - non progressive congenital ataxia: clinical and radiological findings in a pair of siblings. AB - We describe the clinical and radiological findings of a pair of siblings with cerebellar vermis hypoplasia and compare them with the literature. Both of them present pregnancies and deliveries uneventful and both presented some grade of hypotonia, ataxia, ocular motor abnormalities and mild motor delay and slurred speech. These siblings meet many of the criteria described in non-progressive congenital ataxia in which can occur familial cases with cerebellar atrophy, including vermis hypoplasia. As differential diagnosis we compare them with related syndromes and with Joubert's syndrome which main radiological finding on MRI is vermis hypoplasia associated with "molar tooth" appearance. The correct answer for these cases will only be possible by molecular genetics. PMID- 11018829 TI - Palatal myoclonus: report of two cases. AB - We describe two cases of palatal myoclonus (PM), one essential and another secondary to a stroke. Case 1: a 64 years old female who developed clicking sounds in both ears after a stroke and three years later on noticed a progressive involuntary movement of the throat associated with rhythmic contractions of the soft palate, muscles of tongue and throat. MRI showed an ischemic area in brainstem. The patient had a partial response to the use of sumatriptan 6 mg subcutaneously. Case 2: a 66 years old female who began with ear clicking at left ear that worsed slowly associated with tinnitus and arrhythmic movements of soft palate and an audible click at left ear. Brain MRI was normal; audiometry showed bilateral neurosensory loss. She was prescribed clonazepan 1 mg daily with complete recovery. Primary and secondary palatal myoclonus share the same clinical features but probably have different pathophysiological underlying mechanisms. PMID- 11018830 TI - Pneumothorax and pneumoperitoneum during the apnea test: how safe is this procedure? AB - Apnea test is a crucial requirement for determining the diagnosis of brain death (BD). There are few reports considering clinical complications during this procedure. We describe a major complication during performing the apnea test. We also analyse their practical and legal implications, and review the complications of this procedure in the literature. A 54 year-old man was admitted for impaired consciousness due to a massive intracerebral hemorrhage. Six hours later, he had no motor response, and all brainstem reflexes were negative. The patient fulfilled American Academy of Neurology (AAN) criteria for determining BD. During the apnea test, the patient developed pneumothorax, pneumoperitoneum, and finally cardiac arrest. Apnea test is a necessary requirement for the diagnosis of brain death. However, it is not innocuous and caution must be take in particular clinical situations. Complications during the apnea test could be more frequent than reported and may have practical and legal implications. Further prospective studies are necessary to evaluate the frequency and nature of complications during this practice. PMID- 11018831 TI - Neurocysticercosis in a preschool-age child: case report. AB - Neurocysticercosis is a frequent cause of epilepsy and other neurologic abnormalities in all age groups, however by virtue of its prolonged incubation period as well as young children's nutritional habits, it is rarely seen in preschool-age children. The objective of this study is to report the case of a 2 year-old child who presented with new-onset seizures. Her diagnostic features, including neuroradiologic findings, are described and compared with the literature. No matter how young he or she may be, every child who presents with new-onset seizures or other unexplained neurologic features and whose CT or MRI shows cystic lesions or contrast-enhancing rounded lesions should raise a suspicion of neurocysticercosis. PMID- 11018832 TI - Generalized status epilepticus associated with massive pulmonary aspiration and transient central diabetes insipidus: case report. AB - Status epilepticus causes significant morbidity and mortality. A case of generalized status epilepticus followed by massive pulmonary aspiration, acute respiratory failure and transient central diabetes insipidus is presented. Seizures were promptly controlled, but the patient required mechanical ventilation and correction of polyuria with desmopressin acetate. During hospitalization mental status improved, diabetes insipidus spontaneously remitted and he was discharged without neurologic sequelae. The clinical and pathophysiological features of this case are discussed. PMID- 11018833 TI - Epilepsia partialis continua (Koshevnikov): a preliminary case report. AB - We report on the preliminary clinical and electrophysiological aspects of an in patient possibly presenting epilepsia partialis continua (Koshevnikov). We discuss the different etiologies and emphasize on the possible idiopathic form in this case. PMID- 11018834 TI - [Bradycardia as an epileptic manifestation in temporal epilepsy: report of a case]. AB - We describe a patient who had cardiac arrhythmia as epileptic manifestation. In a 34-year-old woman who had many episodes of loss of consciousness, the simultaneous ECG and video-EEG monitoring recorded bradycardia with a short episode of asystole (4 seconds) and left temporal rhythmic theta activity on EEG. MRI showed a small mass lesion in the left parahippocampal gyrus. Alterations in cardiac rhythm have been reported in epileptic seizures and tachycardia is the most common finding associated with them; bradyarrhythmia during seizures was uncommon. Many interconnections among insular cortex, limbic system and hypothalamus, may be responsible for vegetative manifestations in temporal lobe epilepsy. PMID- 11018835 TI - [Implicit memory and its contribution to the rehabilitation of an amnesic patient: case report]. AB - Even severe amnesic patients usually are able to learn new information using implicit memory strategies. In the present study we describe our experience in the rehabilitation of an amnesic patient who had suffered cerebral anoxia. He was taught to use a computer text editor, enabling him to sheltered work. The training program lasted 14 weeks and was based on errorless learning and drilling practice techniques. The patient succeeded in all tasks, even though he could not remember accurately the training sessions. These results meet previous studies which point out that interaction between implicit and residual explicit memory can provide a basis new learning in amnesic patients. The domain-specific knowledge acquisition is demonstrating effectiveness in neuropsychological rehabilitation of brain injured patients. PMID- 11018836 TI - [Third ventricle meningioma in a child: case report]. AB - Meningiomas in childhood are rare, and those located in the third ventricle are even rarer, with only 16 cases reported. A five-year-old girl was seen at our hospital with headache and bilateral papilledema. CT scan showed a large third ventricular mass and associated hydrocephalus. Transcallosal interfornical approach was performed with total removal of the tumor, that was attached to the choroid plexus of the anterior third ventricle. Histologically the lesion proved to be a transitional meningioma. The recovery was uneventful, a permanent shunt was not necessary. The diagnosis of these tumors can be done using CT scan or MRI, that usually are sufficient for surgical planning. Angiography usually is not required, unless a vascular lesion is concerned, or embolization is necessary. The treatment of these tumors is surgical, aiming the total removal of the lesion, usually curative once it is acomplished. We report this case, and add a brief review of the literature on third ventricle meningiomas. PMID- 11018837 TI - [Polymyositis and HTLV-I infection: case report]. AB - We report the case of a 57 years-old woman presenting polymyositis associated to HTLV-I infection without clinical signs of involvement of the central and peripheral nervous system. Pathophysiologic aspects of muscular involvement in HTLV-I infection are discussed. PMID- 11018838 TI - [Endoscopic transepto-interforniceal approach to colloid cysts: case report]. AB - The endoscopic treatment of colloid cysts of third ventricle is still controversial. Management of the patient can include observation and serial CT or MRI imaging only, ventriculo peritoneal shunt only, stereotactic aspiration, microsurgery and endoscopy. The author reports the case of a patient with colloid cyst of third ventricle located in a retroforaminal position, in the diencefalic roof, dissecting the raphe fornix, expanding supero-posteriorly, inside the cavum of the septum pellucidum, protruding on the floor of lateral ventricles. The cyst was approached with a rigid neuroendoscope, through a pre-coronal burr hole, 2.5 cm lateral to the midline (Kochers point). Following a transventricular route, the right leaf of the septum pellucidum was endoscopically opened with bipolar, posterior to the septal vein. A yellowish hard lesion was found and partially resected with biopsy forceps, between the two fornices, which were split apart by the uncommon expansion and location of the lesion, allowing a new endoscopic transventricular transepto-interforniceal approach. We also performed endoscopic septostomy and third ventriculostomy. The microsurgical approaches compared with endoscopic approaches, and the possibility of endoscopic alternative routes suitable for selected cases with superior expansion towards the fornix and septum pellucidum is discussed. PMID- 11018839 TI - [Lipoma of the corpus callosum associated with the hypertrophy of the corpus callosum: case report]. AB - Despite the lipomas are the tumors that more commonly occur in the corpus callosum (CC), its incidence in the population is not common. We report on a 5 year old boy, with history of retardation in the psychomotor development and disturbs in the gait, secondary to generalized hypotony. Magnetic resonance imaging showed a curvilinear lipoma of the CC related to its hypertrophy. In the literature patients with CC lipoma usually have agenesis or hypotrophy of the CC, but in the reported case we have seen, by the first time, a hypertrophy of the CC. We made embryologic, genetic, clinical, radiographic and therapeutic considerations about the patients that have CC lipoma comparing to findings in the case we report. PMID- 11018840 TI - [Lipoma of the cerebellopontine angle: case report]. AB - Lipoma of the cerebellopontine angle is a very rare tumor. We report the case of a 14-years-old female, with left side deafness during three years, associated with headache. CT scan showed an hypodense mass, without enhancement at the cerebellopontine angle. The patient was treated surgically by left retrosigmoid approach. The lesion involved the eighth and seventh cranial nerves and only a partial removal was performed. The postoperative course was uneventful. She had no more headache; the deafness of the left side remained unchanged. Asymptomatic lipoma of the cerebellopontine angle can be treated conservatively, although those with progressive symptoms should be treated surgically, with total or partial remove based on their neurovascular involvement. PMID- 11018841 TI - [Cavernoma of the lateral ventricle: case report]. AB - Cavernous malformations are uncommon lesions that are usually present in the cerebral hemispheres. They occur rarely in the ventricular system, and even more rarely in the lateral ventricle. Only 28 cases have been previously reported in the literature. CASE: We present one case of lateral ventricle cavernoma in a 15 years-old female patient, who suffered of mild chronic headache for 8 months, followed by two episodes of sudden intensive headache and stupor with complete recovery after 48 hours. CT scan was performed and revealed a voluminous size, hiperdense mass in the frontal horn of the lateral ventricle. The surgical access to site was through transcallosal interhemisphere approach. The patient had a good recovery without complications. CONCLUSION: Although lateral ventricle cavernomas are rare they should be considered in the differential diagnosis of intraventricular tumors. A wrong preoperative diagnosis has sometimes induced a wrong therapy, such as radiotherapy, for these surgically curable benign lesions. PMID- 11018842 TI - [Hemichorea-hemiballism associated to cryptococcal granuloma in a patient with AIDS: case report]. AB - Movement disorders are not common in acquired immunodeficiency syndrome. Hemichorea-hemiballism (HC-HB) is the most common of them all, and it is usually related to oportunistic toxoplasmosis of the basal ganglia. We present a 28-year old man, HIV positive with HC-HB caused by a right subthalamic granuloma, which did not respond to treatment for toxoplasmosis. Cryptoccococic antigen was positive in the cerebrospinal fluid and antifungic therapy led to clinical and radiologic improvement, thus the diagnosis of a granulomatous lesion by Cryptococcus neoformans was established. Current literature on HC-HB and its relationship with AIDS is subsequently reviewed. PMID- 11018843 TI - Yes, we have bananas (and also health statistics data): Brazilian-based medicine. PMID- 11018844 TI - Nutritional evaluation and support for children infected with HIV. PMID- 11018845 TI - Detection of human papillomavirus in epithelial lesions of the conjunctiva. AB - CONTEXT: Many factors like exposure to UV radiation, climatic conditions, genetic predisposition, immunological state and, more recently, the presence of HPV have been implicated in the genesis of some lesions of the conjunctiva, especially the carcinoma. OBJECTIVE: To evaluate the presence of HPV DNA in acquired lesions of the conjunctiva and in normal mucosa. TYPE OF STUDY: Cross-sectional study. SETTING: A public university referral center (the Ophthalmology Service of the A.C. Camargo Hospital - A. Prudente Foundation, Sao Paulo). PARTICIPANTS: Thirty patients with acquired lesions of the conjunctiva and 60 matched controls (by age and sex) were evaluated in this study, from June 1993 to March 1995. PROCEDURES: The detection of HPV DNA in the normal conjunctiva and in acquired lesions was done by the PCR technique and dot blot hybridization. The material was collected by scraping the normal mucosa and the surface of the lesions. A fragment of fresh frozen tissue and paraffin embedded specimens of each lesion were also included. MAIN MEASUREMENTS: The association between the HPV infection and the presence or absence of conjunctival lesions. RESULTS: Sequences of HPV DNA were detected in 4 of the 31 lesions evaluated (12.9%) and in the healthy mucosa of one individual of the control group (1.6%). HPV type 16 was detected in 2 carcinomas and in the normal mucosa of one individual of the control group. HPV type 11 was demonstrated in 2 papillomas of one patient with lesions in both eyes. CONCLUSIONS: The low frequency of HPV DNA found in the lesions of this sample and the detection of the viral genome in the normal mucosa indicate that there is a weak possibility of association between HPV infection and the carcinoma of the conjunctiva. PMID- 11018846 TI - Epidemiology of bloodstream infections at a cancer center. AB - CONTEXT: Cancer patients are at unusually high risk for developing bloodstream infections (BSI), which are a major cause of in-hospital morbidity and mortality. OBJECTIVE: To describe the epidemiological characteristics and the etiology of BSI in cancer patients. DESIGN: Descriptive study. SETTING: Terciary Oncology Care Center. PARTICIPANTS: During a 24-month period all hospitalized patients with clinically significant BSI were evaluated in relation to several clinical and demographic factors. RESULTS: The study enrolled 435 episodes of BSI (349 patients). The majority of the episodes occurred among non-neutropenic patients (58.6%) and in those younger than 40 years (58.2%). There was a higher occurrence of unimicrobial infections (74.9%), nosocomial episodes (68.3%) and of those of undetermined origin (52.8%). Central venous catheters (CVC) were present in 63.2% of the episodes. Overall, the commonest isolates from blood in patients with hematology diseases and solid tumors were staphylococci (32% and 34.7%, respectively). There were 70 episodes of fungemia with a predominance of Candida albicans organisms (50.6%). Fungi were identified in 52.5% of persistent BSI and in 91.4% of patients with CVC. Gram-negative bacilli prompted the CVC removal in 45.5% of the episodes. Oxacillin resistance was detected in 26.3% of Staphylococcus aureus isolates and in 61.8% of coagulase-negative Staphylococcus. Vancomycin-resistant enterococci were not observed. Initial empirical antimicrobial therapy was considered appropriate in 60.5% of the cases. CONCLUSION: The identification of the microbiology profile of BSI and the recognition of possible risk factors in high-risk cancer patients may help in planning and conducting more effective infection control and preventive measures, and may also allow further analytical studies for reducing severe infectious complications in such groups of patients. PMID- 11018847 TI - The frequency of smoking and problem drinking among general hospital inpatients in Brazil - using the AUDIT and Fagerstrom questionnaires. AB - CONTEXT: Although the CAGE questionnaire is one of the most widely used alcohol screening instruments, it has been criticized for not identifying people who are drinking heavily or who have alcohol related problems but do not as yet show symptoms of alcohol dependence. The AUDIT (Alcohol Use Disorder Identification Test) questionnaire was developed by WHO as a screening instrument specifically designed to identify problem drinkers, as well as those who were already dependent on alcohol. OBJECTIVE: The aim of this study was to use the AUDIT and Fagerstrom questionnaires in a general hospital inpatient population to measure the frequency of problem drinking and nicotine dependence, and to see if levels varied between medical speciality. DESIGN: Retrospective cross-sectional study. SETTING: Federally funded public teaching hospital. SAMPLE: 275 inpatients from both genders. MAIN MEASUREMENTS: Socio-demographic data, AUDIT (Alcohol Use Disorders Identification Test) and Fagerstrom Test for Nicotine Dependence. RESULTS: We interviewed 275 inpatients, 49% of whom were men and 51% women. Thirty-four patients were identified as "cases" by the Audit questionnaire; 22% of the male patients and 3% of the females. Just over 21% of inpatients were current smokers. The gastroenterology (26%) and general medicine (16%) inpatient units had the largest number of individual cases. CONCLUSIONS: Only by knowing the prevalence of alcohol abuse/dependence and nicotine dependence in a general hospital can we evaluate the need for a specialized liaison service to identify and treat these patients. PMID- 11018848 TI - Thyroid stimulating hormone levels in cord blood are not influenced by non thyroidal mothers' diseases. AB - CONTEXT: Screening programs not only offer the opportunity to trace and treat almost all cases of congenital hypothyroidism but also mean large savings to the health system. However, carefully planned strategies are necessary to extend their benefits and reduce costs. OBJECTIVE: To determine the possible influence of maternal diseases that affect maternal-fetal placenta dynamics on primary thyroid stimulating hormone (TSH) screening for congenital hypothyroidism. DESIGN: Prospective non-randomized clinical trial with at least 3 months of follow-up. SETTING: A public university referral center [CAISM/Hospital das Clinicas, Faculty of Medicine, University of Campinas, Campinas, SP]. PARTICIPANTS: 415 neonates divided into 5 groups: eighty-three infants born from cardiac mothers; 98 from mothers that had toxemia; 54 of the mothers had diabetes mellitus; 40 were HIV positive and 140 had no diseases. INTERVENTION: All newborns had cord blood samples collected on filter paper at birth. MAIN MEASUREMENTS: TSH was measured from dried blood spots using a homemade immunofluorescence assay (sensitivity in dried blood spots = 0.1 mU/L). RESULTS: There was no significant difference in the mean TSH levels among the 5 groups. Moreover, TSH levels were around 5 mU/L in 48% of the newborns, indicating that our region is severely deficient in iodine. CONCLUSIONS: Our results indicate that primary TSH screening programs using cord blood are not affected by maternal diseases. We suggest that, besides its technical advantages over heel punctures with T4 primary approaches, neonatal screening using primary cord blood TSH may also be used as a monitoring tool for evaluation and control of iodine deficiency disorders (IDD). PMID- 11018849 TI - Evolution of nutritional status of infants infected with the human immunodeficiency virus. AB - CONTEXT: There are today only a limited number of studies defining growth parameters and nutritional status for HIV children. OBJECTIVE: To study the nutritional status of infants infected with the human immunodeficiency virus. TYPE OF STUDY: Longitudinal study. SETTING: Department of Pediatrics, Faculty of Medical Sciences, UNICAMP, Campinas, Brazil. PARTICIPANTS: One hundred and twenty four children born to HIV infected mothers were evaluated from birth until the age of two years. They were subdivided into two groups: 71 infected children and 53 non-infected children. MAIN MEASUREMENTS: Growth was evaluated in both groups by comparing Z-scores for weight/age (w/a), length/age (H/a) and weight/length (w/H) (using the NCHS curves as reference). RESULTS: The Z-score analyses showed that there was a significant difference between the two groups for all the variables studied, except for the H/a value at 3 months of age and the W/H value at 21 months of age, which showed P > 0.05. CONCLUSIONS: The growth of infected infants was observed to be severely affected in comparison with that of seroreversed infants in the same age groups. Although clinical manifestations may take time to appear, the onset of growth changes begin soon after birth. PMID- 11018850 TI - Vanishing bile duct syndrome in Hodgkin's disease: case report. AB - CONTEXT: Liver damage is relatively common in patients affected by Hodgkin's disease. A smaller proportion of cases develops jaundice. Recently, the vanishing bile duct syndrome was described in Hodgkin's disease. The mechanisms of this severe complication have been poorly understood until now. OBJECTIVE: To describe a rare case of intra-hepatic cholestasis due to vanishing bile duct syndrome. DESIGN: Case report. CASE REPORT: A 38-year-old male patient affected by Hodgkin's disease. Liver biopsy showed no detectable Hodgkin's disease. Intra hepatic cholestasis was found and none of the six portal tracts analyzed contained normal bile ducts. The treatment was based on conventional and high dose escalation chemotherapy. The patient died from an irreversible liver failure while in complete remission from Hodgkin's disease. PMID- 11018851 TI - Musculoskeletal manifestations of bacterial endocarditis. AB - CONTEXT: The incidence of staphylococcal infection has been increasing during the last 20 years. OBJECTIVE: Report a case of staphylococcal endocarditis preceded by musculoskeletal manifestations, which is a rare form of clinical presentation. DESIGN: Case report. CASE REPORT: A 45-year-old-man, without addictions and without known previous cardiopathy, was diagnosed as having definitive acute bacterial endocarditis due to Staphylococcus aureus. Its etiology was community acquired, arising from a non-apparent primary focus. In addition, the musculoskeletal symptoms preceded the infective endocarditis (IE) by about 1 month, which occurred together with other symptoms, e.g. mycotic aneurysms and petechiae. Later, the patient showed perforation of the mitral valve and moderate mitral insufficiency with clinical control. PMID- 11018852 TI - Comparative analysis of HIV type 1 genotypic resistance across antiretroviral trial treatment regimens. AB - From data on HIV-1 genotypes collected from antiretroviral trial participants who fail virologically, we describe methods for comparing distributions of acquired HIV-1 mutations across different treatment regimens. Given a definition of a "mutational distance" that summarizes the genetic change of a subject's virus in a way that captures the resistance cost of exposure to an antiretroviral regimen, these comparative analyses inform about the relative treatability of emergent virus by next-line therapy directed to the same viral target. The utility of the methods is illustrated by application to data from AIDS Clinical Trials Group (ACTG) Study 241. We find that patients failing zidovudine/didanosine/nevirapine accumulated a 2.41-fold greater nonnucleoside reverse transcriptase inhibitor (RTI) mutational distance than patients failing zidovudine/didanosine [95% confidence interval (1.55, 5.26), p < 0.000001], quantitating expectations that adding a nonnucleoside RTI to a double nucleoside regimen may attenuate future effectiveness of nonnucleoside RTI therapy for nucleoside-experienced patients if viremia is not suppressed. We also find that persons with extensive prior experience with suboptimal nucleoside therapy who were virologically failing zidovudine/didanosine/nevirapine or zidovudine/didanosine accumulated a similar nucleoside RTI mutational distance, implying that the addition of the nonnucleoside RTI did not preserve future nucleoside options. PMID- 11018853 TI - Effect of lamivudine in HIV-infected persons with prior exposure to zidovudine/didanosine or zidovudine/zalcitabine. AB - Nucleoside analog-based regimens remain an integral component of combination therapy for use in both antiretroviral treatment-naive and experienced HIV infected patients. To further define treatment responses to new antiretroviral therapy in patients with long-term experience to dual nucleoside analog therapy (zidovudine [ZDV] plus didanosine [ddI] or ZDV plus zalcitabine [ddC]), 325 subjects derived from the AIDS Clinical Trials Group (ACTG) 175 trial were randomized to three different combination regimens: (1) continuation of ZDV + ddI or ZDV + ddC (continuation arm), (2) addition of 3TC to ZDV + ddI or ZDV + ddC (addition arm), or (3) a switch to ZDV + 3TC therapy (switch arm). Both the addition and switch arms sustained significantly greater short-term (baseline to week 4) mean CD4+ cell count increases compared with the continuation arm (+36, +28 versus -4 cells/mm3; p = 0.012) and long-term CD4+ cell count responses (baseline to weeks 40/48: +32, +19 versus -9 cells/mm3; p = 0.003). Superior short-term (baseline to week 8) mean decreases in plasma HIV RNA (p < 0.001) were achieved by both the addition and switch arms (0.53 log10 and 0.54 log10 copies/ml, respectively) compared with the continuation arm (0.13 copies/ml) whereas no differences in long-term virologic suppression were observed (p = 0.30). At week 48, no differences were observed in the proportions of subjects who had HIV RNA levels below 500 copies/mL: 18% of subjects in each treatment arm (3-way p = 1.0). Overall, the treatments were well tolerated and only nine subjects (3%) died or developed one or more AIDS-defining events. While this study confirms the intrinsic antiretroviral activity of 3TC, only modest marker changes and limited short-term viral suppression are seen with incremental addition of the drug. The current approach of using 3TC in maximally suppressive regimens is preferred. PMID- 11018854 TI - Thalidomide for the treatment of AIDS-associated wasting. AB - A double-blind, placebo-controlled trial of efficacy and safety of thalidomide in AIDS-associated wasting was carried out. Ninety-nine of 103 male patients had at least one on-study measurement (intent-to-treat [ITT] cohort). Patients were randomized to thalidomide at 100 mg/day (T100) or 200 mg/day (T200), or placebo for 8 weeks. By ITT analysis, the mean change in body weight of the placebo, T100, and T200 treatment groups was 0.3 kg (0.4%), 2.0 kg (3.0%), and 0.9 kg (1.4%), respectively (p = 0.021 for T100 versus placebo; p = 0.53 for T200 versus placebo). Of the 64 patients who completed the 8 weeks of study treatment, significant weight gain was observed in both the T100 group (2.2 kg, [33%]; p = 0.008 versus placebo) and the T200 group (1.5 kg [2.5%]; p = 0.019 versus placebo). Approximately half the weight gain was fat-free mass (bioimpedance analysis). Patients in the T100 or T200 groups had no significant change in CD4+ cell counts, neutrophil counts, or TNF-alpha levels, compared with placebo. HIV viral load measured as log10 copies/ml decreased by a median of 0.07 in the placebo group, and increased by a median of 0.29 (T100 group) and 0.23 (T200 group) (p = 0.024 andp = 0.018 versus placebo, respectively). Thalidomide therapy was associated with mild to moderate rashes and fevers, but not peripheral neuropathy. Although the anabolic benefits of high-dose thalidomide are limited by drug intolerance, 8 weeks of low-dose thalidomide results in significant weight gain in patients with AIDS-associated wasting. PMID- 11018855 TI - Highly active antiretroviral therapy results in HIV type 1 suppression in lymph nodes, increased pools of naive T cells, decreased pools of activated T cells, and diminished frequencies of peripheral activated HIV type 1-specific CD8+ T cells. AB - This study examines sequential lymph nodes from 13 drug-naive patients before and after 24 weeks of highly active antiretroviral therapy (HAART). A multipronged approach was used to study changes in HIV-1 RNA in each paired lymph node in relation to tissue architecture and frequency of naive T cells. After 24 weeks, all patients showed significant suppression of plasma viral load and 12 of 13 showed concordant viral suppression in the lymph node (p = 0.001). Using in situ hybridization and quantitative image analysis, we showed that HIV-1 RNA was reduced to below detectable levels (two copies per cell) in follicular dendritic cell (FDC) and mononuclear cell pools. Independent immunohistochemical analysis of lymph node sections revealed that 5 of 13 patients displayed increased FDC networks and 6 of 13 showed no change and all patients showed increases in tissue resident CD4+ cells. All lymph node biopsies at 24 weeks showed increased proportions of CD4+ and CD8+ cells coexpressing the naive markers CD45RA and CD62L when compared with baseline values. Significant correlations existed between viral load suppression and loss of activated CD8+ T cells after 24 weeks in both lymph node and blood, which was mirrored by significantly lowered frequencies of activated peripheral Gag peptide/MHC tetramer+ CD8+ cells. Overall, these data show that a potent and successful treatment strategy that significantly suppresses and removes FDC-resident HIV-1 results in improvements in lymphoid architecture and by so doing provides the structures available for increased numbers of naive cells to interact with cognate antigen. In addition, our article shows that suppression of HIV-1 replication results in diminished frequencies of peripherally activated antigen-specific CD8+ cells. PMID- 11018856 TI - Dual infection with human immunodeficiency virus type 1 and type 2: impact on HIV type 1 viral load and immune activation markers in HIV-seropositive female sex workers in Abidjan, Ivory Coast. AB - To determine the impact of dual infection with HIV-1 and HIV-2 on HIV-1 viral load and markers of immune activation among HIV-seropositive FSWs in Abidjan, we analyzed blood samples obtained from consenting HIV-seropositive FSWs attending a confidential clinic between September 1996 and June 1997 in Abidjan. Among HIV-1 and HIV-2 dually seropositive FSWs, polymerase chain reaction (PCR) testing with HIV-1 and HIV-2 primers was used to differentiate between FSWs who were PCR positive only for HIV-1 and those positive for both HIV-1 and HIV-2 (dually infected). Of the 203 FSWs, 151 (74%) were HIV-1 seropositive only (median age, 26 years), 4 (2%) were HIV-2 seropositive, and 48 (24%) were dually seropositive (median age, 30 years). Of the 48 dually seropositive FSWs, 33 (69%) were dually infected and 15 (31%) were dually seropositive. Median CD4+ T cell counts per microliter were not significantly different among the three groups (525 for HIV-1 positive only, 502 for dually infected, and 416 for dually seropositive) (p = 0.14). Median viral load (log10 copies/ml) was not significantly different among the HIV-1-only FSWs (4.8 log10 copies/ml) compared with the 32 dually infected FSWs (4.6 log10 copies/ml) and 14 dually seropositive FSWs (4.7 log10 copies/ml; p = 0.95). Median levels of HLA-DR immune activation were increased in both CD4+ and CD8+ T cells for the dually infected (n = 27) FSWs compared with those infected with HIV-1 only (n = 123) (p = 0.019 and p = 0.01, respectively). Dual infection does not appear to influence levels of HIV-1 viral load in vivo. However, levels of HLA-DR are higher among FSWs dually infected with HIV-1 and HIV-2 than among those infected with HIV-1 only. PMID- 11018857 TI - High plasma levels of soluble fas in HIV type 1-infected subjects are not normalized during highly active antiretroviral therapy. AB - Plasma levels of soluble Fas (sFas) are elevated in human immunodeficiency virus type 1 (HIV-1) infection, indicating dysregulation of the Fas apoptosis pathway and chronic immune activation. We performed a retrospective study to investigate the effects of HAART on plasma levels of sFas. A cross-sectional study of 27 drug naive infected subjects and 49 patients under antiretroviral treatment showed that plasma levels of sFas were higher in HIV-1-infected subjects than in 52 HIV 1-negative controls, independently of the treatment status. In a longitudinal study of 69 patients undergoing HAART, we observed a minimal, but significant decrease in sFas plasma levels after 1 year of therapy. Levels of sFas, however, remained still higher than physiologic values. Patients undergoing HAART were further classified as nonresponders or responders on the basis of viremia suppression; no significant changes in plasma levels of sFas were observed between the two groups. These findings show that 1 year of HAART has a minor effect on the sFas levels in plasma. Long-term HAART may be required to normalize the dysregulation of the Fas apoptotic pathway and the persistent immune activation initiated by HIV-1. PMID- 11018858 TI - Natural residues versus antiretroviral drug-selected mutations in HIV type 1 group O reverse transcriptase and protease related to virological drug failure in vivo. AB - HIV-1 group O viruses were first recognized as a distinct subgroup of HIV-1 with the isolation and characterization in 1990 of a virus (ANT70) from a woman (individual A) and her spouse (individual B), both from Cameroon (De Leys R, et al.: J Virol 1990;64:1207-1216). During the 5-6 years before treatment, individual A remained asymptomatic, with viral RNA levels between 2.5 and 2.8 log10 copies/ml, as measured by a newly developed group O-specific quantitative NASBA-based RNA assay. Individual B developed mild clinical symptoms, with 3.1 to 3.6 log10 copies of viral RNA per milliliter. HIV-1 sequences obtained from both individuals showed pretreatment residues in protease that confer resistance to protease inhibitors in group M viruses (10I, 36I, and 71V). Individual A showed an initial response to AZT, but shortly after addition of ddC and saquinavir, the RNA levels returned to baseline, while subsequent treatment with d4T, 3TC, and indinavir reduced the RNA level to less than 50 copies/ml for the time of follow up. Individual B showed no response to AZT or ddC monotherapy, and a change to d4T, 3TC, and indinavir had, in contrast to individual A, only a temporary effect. While a multitude of mutations in HIV-1 group O reverse transcriptase (RT) and protease appeared that are associated with drug resistance in group M viruses, the observed T215N mutation in RT and the V15I and V22A mutations in protease have not previously been described and may represent resistance conferring mutations specific to group O viruses. These results indicate that treatment of HIV-1 group O-infected individuals with antiretroviral drug regimens that include protease inhibitors might lead to rapid selection for resistance conferring mutations. This probably results from preexisting protease residues contributing to reduced sensitivity of group O viruses to protease inhibitors, as is observed in vitro. PMID- 11018859 TI - Protease gene analysis of HIV type 1 non-B subtypes in Spain. AB - The protease gene of human immunodeficiency virus type 1 (HIV-1) clinical isolates found in 15 immigrants (most of African origin) living in Spain was examined. Phylogenetic analyses were performed, taking as reference a panel of 26 HIV protease gene sequences deposited with GenBank. All specimens belonged to four distinct HIV-1 non-B subtypes: C (three cases), F (one), G (nine), and H (two). Five patients harboring subtype G strains were further classified within the IbNg recombinant clade. A high degree of genetic polymorphism at the protease gene was seen in all subtypes. Moreover, changes at positions associated with drug resistance were seen in subtype G viruses carried by patients who had not been exposed to protease inhibitors. Plasma viremia was lower than expected for some samples, according to the clinical features and the CD4+ cell count, suggesting that viral load titers were underestimated by all three commercially available techniques. This work represents the first genetic characterization of subtypes C, F, G, and H in Spain. PMID- 11018860 TI - Strain-dependent productive infection of a unique eosinophilic cell line by human immunodeficiency virus type 1. AB - Eosinophils are granulocytic leukocytes that function in both protective and pathological immune responses. They can be infected by HIV-1, but characterization of the infection has been hindered by lack of a productive cell culture model. In the present study, the unique eosinophilic cell line AML14.3D10 was used as a model to test the hypothesis that HIV-1 productively infects eosinophilic cells in a strain-dependent fashion. The AML14.3D10 cell line was cultured with one T cell-tropic (T-tropic) strain and two macrophage-tropic (M tropic) strains of HIV-1 (HTLV-IIIB, HIV-1AdaM, and HIV-1Ba-L strains, respectively). Cytopathic effects were evident in living cultures and in stained slide preparations of AML14.3D10 cells infected with the T-tropic strain of HIV 1. Culture supernatants from infected AML14.3D10 cells contained high levels of HIV-1 p24 protein that peaked at approximately 7-10 days postinfection. A line of AML14.3D10 cells chronically infected with HTLV-IIIB and continuously producing high levels of virus was established. In contrast to the T-tropic strain, the M tropic strains of HIV-1 did not productively infect the eosinophilic cell line. Thus, the AML14.3D10 eosinophilic cell line was permissive for a T-tropic strain but not for M-tropic strains of HIV-1. Flow cytometry revealed that uninfected AML14.3D10 cells were positive for the HIV-1 receptor CD4 and coreceptors CXCR4 and CCR5; the cell line was negative for CCR3. The lack of productive infection by M-tropic strains despite CCR5 expression indicates that strain-dependent infection may not be determined at the coreceptor level in AML14.3D10 cells. PMID- 11018861 TI - Mutations in the HIV type 1 integrase of patients receiving long-term dideoxynucleoside therapy do not confer resistance to zidovudine. AB - Metabolites of AZT can inhibit HIV-1 integrase in vitro (Mazumder A, et al., Proc Natl Acad Sci USA 1994;91:5771-5775). To determine if long-term dideoxynucleoside therapy can lead to the emergence of HIV-1 AZT-resistant variants containing mutations in the integrase, we have sequenced the proviral DNA encoding the HIV-1 integrase of nine HIV-1-infected patients at different time points during treatment. Four of the nine patients developed mutations during the course of treatment. Although most mutations occurred at nonconserved amino acids, one patient developed a mutation at codon (R166T), a residue that is conserved among all integrases from known HIV-1 isolates. This mutation was introduced in the recombinant HIV-1 integrase protein to determine if it could confer resistance to AZT in vitro. We show that the R166T integrase mutant is still proficient at carrying 3'-processing and 3' end-joining but that the enzyme is not resistant to AZT-TP. Our results suggest that it is unlikely that integrase inhibition contributes to the antiviral activity of AZT. PMID- 11018862 TI - Plasma and salivary IgG subclasses in HIV type 1 infection: evidence of both transudation and local synthesis of IgG in parotid saliva. AB - In this study we measured the levels of plasma and salivary IgG subclasses in 81 HIV-1-infected individuals and 30 uninfected controls. Salivary IgG1 was increased in HIV-1-infected patients, while salivary IgG2 and IgG4 were decreased. Patients with high levels of plasma anti-HIV-1 IgG antibodies presented a higher CD4+ cell count and lower viral load. High levels of anti-HIV 1 IgG antibodies in plasma were also associated with high levels of anti-HIV-1 IgG antibodies in parotid saliva. In comparing the HIV-1 recognition patterns of salivary IgG with plasma IgG, we determined that plasma and salivary IgG antibodies had similarities as well as differences in their reactivity to HIV-1 antigens. The present study demonstrates that HIV-1 infection affects both plasma and salivary IgG and provides evidence that the origin of HIV-1-specific IgG antibodies in parotid saliva is primarily transudated from plasma; however, some local synthesis of IgG in parotid saliva also occurs. PMID- 11018863 TI - Identification of highly conserved and broadly cross-reactive HIV type 1 cytotoxic T lymphocyte epitopes as candidate immunogens for inclusion in Mycobacterium bovis BCG-vectored HIV vaccines. AB - One of the fundamental goals of current strategies to develop an efficacious vaccine for AIDS is the elicitation of cytotoxic T lymphocyte (CTL) reactivities capable of recognizing cells infected with different subtypes of the human immunodeficiency virus type 1 (HIV-1). In efforts to explore new vaccine candidates by the UNAIDS/WHO Vaccine Committee, we review the most recent data concerning CTL epitopes that are conserved among the different HIV-1 subtypes. Moreover, we examine HLA allelic frequencies in several different populations, to determine those that could contribute to the goal of a cumulative phenotype frequency (CP) of at least 80%. By analyzing conserved epitopes in the context of HLA restricting alleles, we define a set of HIV-1 gene regions that may have the greatest potential to induce cross-clade reactive CTLs. The absence of well defined correlates of immune protection that link CTL epitopes to delayed disease progression and/or prevention of infection does not permit an assignment of rank order of the most relevant component of a candidate vaccine. Thus far, most of the studies conducted in clade B-infected patients to define conserved and immunodominant epitopes indicate gag and pol gene products to be the most conserved among the HIV-1 subtypes. Moreover, anti-Pol and -Gag CTL responses appear to correlate inversely with disease progression, suggesting that they should be among the first choice of antigens to be included in a candidate vaccine construct aimed at induction of broad CTL responses. The impact of a clade B-based vaccine as a worldwide candidate capable of inducing protective immune responses can be determined only after "in vivo" studies. Meanwhile, extensive parallel studies in populations infected with non-clade B HIV-1 subtypes should define the patterns of immunodominant epitopes and HLA for comparison with the data already collected in clade B-infected subjects. PMID- 11018864 TI - A comparison of full-length glycoprotein 120 from incident HIV type 1 subtype E and B infections in Bangkok injecting drug users with prototype E and B strains that are components of a candidate vaccine. AB - Complete gp120 sequence information was obtained from eight persons with incident HIV-1 infections (four subtype E and four subtype B) who were part of a prospective injecting drug user (IDU) cohort in Bangkok, Thailand, during 1996 1998. The incident subtype E strains were similar to the prototype subtype E strain CM244 isolated in 1992 in northern Thailand. The incident subtype B strains displayed divergence, in both overall genetic distance and other significant gp120 characteristics, from the prototype North American subtype B strain HIV-MN. Recombinant gp120s derived from CM244 and HIV-MN strains are components of a vaccine that is undergoing phase III efficacy testing, begun in March 1999, among Bangkok area IDUs. The information presented here will be important in the evaluation of any breakthrough HIV-1 infections occurring among vaccinees during the vaccine trial and in ongoing vaccine development efforts in Thailand. PMID- 11018865 TI - Human isotretinoin metabolism during indinavir therapy. PMID- 11018866 TI - Minimal potentials with very many minima AB - We demonstrate, by construction, that simple renormalizable matrix potentials with S(N), as opposed to O(N), symmetry can exhibit an exponentially large number of inequivalent deep local minima. PMID- 11018867 TI - Fast time-evolution method for dynamical systems AB - A fast time-evolution method is developed for systems for which the dynamical behavior can be reduced to the eigenvector/eigenvalue problem. The method does not use the eigenvectors/eigenvalues themselves and is based on a polynominal expansion of the formal operator solution in the eigenfrequency domain. It is complementary to the standard time-integration approaches and allows one to calculate or simulate the state of a system at arbitrary times. The time evolution of, e.g., classical harmonic atomic systems and quantum systems described by linear Hamiltonians can be treated by this method. PMID- 11018868 TI - Nonlinear effects in interference of bose-einstein condensates AB - Nonlinear effects in the interference of Bose-Einstein condensates are studied using exact solutions of the one-dimensional nonlinear Schrodinger equation, which is applicable when the lateral motion is confined or negligible. With the inverse scattering method, the interference pattern is studied as a scattering problem with the linear Schrodinger equation, whose potential is profiled by the initial density distribution of the condensates. Our theory not only provides an analytical framework for quantitative predictions for the one-dimensional case, it also gives an intuitive understanding of some mysterious features of the interference patterns observed in experiments and numerical simulations. PMID- 11018869 TI - Motion of dark solitons in trapped bose-einstein condensates AB - We use a multiple time scale boundary layer theory to derive the equation of motion for a dark (or grey) soliton propagating through an effectively one dimensional cloud of Bose-Einstein condensate, assuming only that the background density and velocity vary slowly on the soliton scale. We show that solitons can exhibit viscous or radiative acceleration (antidamping), which we estimate as slow but observable on experimental time scales. PMID- 11018870 TI - General structure of bose-einstein condensates with arbitrary spin AB - Motivated by the recent discoveries of spin-1 and pseudo-spin-1/2 Bose gas, we have studied the general structure of the Bose gases with arbitrary spin. A general method is developed to uncover the elementary building blocks of the angular momentum eigenstates, as well as the relations (or interactions) between them. Applications of this method to Bose gas with integer spins ( f = 1,2,3) and half integer spins ( f = 1/2,3/2) reveal many surprising structures. PMID- 11018871 TI - Condensate statistics in interacting and ideal dilute bose gases AB - We obtain analytical formulas for the statistics, in particular, for the characteristic function and all cumulants, of the Bose-Einstein condensate in dilute weakly interacting and ideal equilibrium gases in the canonical ensemble via the particle-number-conserving operator formalism of Girardeau and Arnowitt. We prove that the ground-state occupation statistics is not Gaussian even in the thermodynamic limit. We calculate the effect of Bogoliubov coupling on suppression of ground-state occupation fluctuations and show that they are governed by a pair-correlation, squeezing mechanism. PMID- 11018872 TI - Suprathreshold stochastic resonance in multilevel threshold systems AB - A new form of stochastic resonance (SR) that occurs in multilevel threshold signal detectors is reported. In contrast to classical SR, which extends the dynamic range of threshold detectors to subthreshold signal levels, this new form of SR extends the dynamic range to suprathreshold signal strengths. The effect is most dominant, and can outperform networks based on standard engineering design, when all thresholds adapt to the dc level of the signal. This has an interesting analogy to dc adaptation in neurons. The possible connection between these two effects is discussed. PMID- 11018873 TI - Consistent canonical quantization of general relativity in the space of vassiliev invariants AB - We present a quantization of the Hamiltonian and diffeomorphism constraint of canonical quantum gravity in the spin network representation. The novelty consists in considering a space of wave functions based on the Vassiliev invariants. The constraints are finite, well defined, and reproduce at the level of quantum commutators the Poisson algebra of constraints of the classical theory. A similar construction can be carried out in 2+1 dimensions leading to the correct quantum theory. PMID- 11018874 TI - Quantum gravity corrections to neutrino propagation AB - Massive spin-1/2 fields are studied in the framework of loop quantum gravity by considering a state approximating, at a length scale L much greater than Planck length l(P), a spin-1/2 field in flat spacetime. The discrete structure of spacetime at l(P) yields corrections to the field propagation at scale L. Neutrino bursts ( &pmacr; approximately 10(5) GeV) accompanying gamma ray bursts that have traveled cosmological distances L are considered. The dominant correction is helicity independent and leads to a time delay of order (&pmacr;l(P))L/c approximately 10(4) s. To next order in &pmacr;l(P), the correction has the form of the Gambini and Pullin effect for photons. A dependence L(-1)(os) approximately &pmacr;(2)l(P) is found for a two-flavor neutrino oscillation length. PMID- 11018875 TI - Multi-TeV scalars are natural in minimal supergravity AB - For a top quark mass fixed to its measured value, we find natural regions of minimal supergravity parameter space where all squarks, sleptons, and heavy Higgs scalars have masses far above 1 TeV and are possibly beyond the reach of the Large Hadron Collider at CERN. This result is simply understood in terms of "focus point" renormalization group behavior and holds in any supergravity theory with a universal scalar mass that is large relative to other supersymmetry breaking parameters. We highlight the importance of the choice of fundamental parameters for this conclusion and for naturalness discussions in general. PMID- 11018876 TI - New approach to axial coupling constants in the QCD sum rule AB - We derive new QCD sum rules for the axial coupling constants by considering the correlation functions of axial-vector currents in a one-nucleon state. The QCD sum rules tell us that the axial coupling constants are expressed by nucleon matrix elements of quark-gluon composite operators which are related to the sigma terms and the moments of parton distributions. The results for the isovector axial coupling constants and the eighth component of the SU(3)(f) octet are in good agreement with experiment. PMID- 11018877 TI - Interference in exclusive vector meson production in heavy-Ion collisions AB - Vector mesons are produced copiously in peripheral relativistic heavy-ion collisions. Virtual photons from one ion can fluctuate into quark-antiquark pairs and scatter from the second ion, emerging as vector mesons. The emitter and target are indistinguishable, so emission from the two ions will interfere. Vector mesons have negative parity so the interference is destructive, reducing the production of mesons with small transverse momentum. The mesons are short lived, and decay before emission from the two ions can overlap. However, the decay-product wave functions overlap and interfere since they are produced in an entangled state, providing an example of the Einstein-Podolsky-Rosen paradox. PMID- 11018878 TI - Elastic compton scattering from the deuteron and nucleon polarizabilities AB - Cross sections for elastic Compton scattering from the deuteron were measured over the laboratory angles straight theta(gamma) = 35 degrees -150 degrees. Tagged photons in the laboratory energy range E(gamma) = 84-105 MeV were scattered from liquid deuterium and detected in the large-volume Boston University NaI spectrometer. Using the calculations of Levchuk and L'vov, along with the measured differential cross sections, the isospin-averaged nucleon polarizabilities in the deuteron were estimated. A best-fit value of (alpha; beta;) = 2.6+/-1.8 was determined, constrained by dispersion sum rules. This is markedly different from the accepted value for the proton of (alpha;-beta;)(p) = 10.0+/-1.5+/-0.9. PMID- 11018879 TI - Out of plane measurements of the decay neutron from the giant resonance in the 12C(e,e(')n)11C reaction AB - Out of plane measurements of the angular correlations for the 12C(e, e(')n) reaction have been performed for the first time in the giant resonance region. The cross sections were directly separated into the longitudinal and transverse, longitudinal-transverse, and transverse-transverse components. The cross section at the peak of the giant resonance ( omega = 22.5 MeV) has been found to be almost all longitudinal. It was reproduced by the multipole expansion with E0 and E2 components besides E1. The longitudinal-transverse component might have a maximum around 24 MeV. The transverse-transverse component is very small over the giant resonance. PMID- 11018881 TI - Atomic quadrupolar photoemission asymmetry parameters from a solid state measurement AB - A method for measuring the asymmetry parameter gamma arising from dipole quadrupole interference in core-level photoemission is proposed which is based on condensed thin films and exploits the influence of this asymmetry in photoemission monitoring of x-ray standing wave field absorption. The high density of material in this method offers some advantages over a conventional gas phase measurement. Results are presented for C, O, and F 1s photoemission at photon energies around 3 keV. The combined nondipole asymmetry factor (delta+gamma/3) for non- s-states can be determined in a similar way. PMID- 11018880 TI - Large enhancement of the sub-barrier fusion probability for a halo nucleus AB - The fusion-fission cross sections of the 4He+238U and 6He+238U systems have been measured, at Louvain-la-Neuve, for energies around and below the Coulomb barrier, using an array of Si detectors surrounding a UF4 target. The data taken with 4He are in good agreement with previous data and with the coupled channel fusion calculation performed with ECIS. The 6He data show a regular trend with a large enhancement below the barrier which is attributed to the halo structure of the 6He nucleus. PMID- 11018882 TI - Density shift and broadening of transition lines in antiprotonic helium AB - The density shift and broadening of the transition lines of antiprotonic helium have been evaluated in the impact approximation using an interatomic potential calculated ab initio with the symmetry-adapted perturbation theory. The results help to remove an uncertainty of up to 10 ppm in the laser spectroscopy data on antiprotonic helium and are of importance in experimental tests of bound state QED and CPT invariance. PMID- 11018883 TI - Tomographic measurement of joint photon statistics of the twin-beam quantum state AB - We report the first measurement of the joint photon-number probability distribution for a two-mode quantum state created by a nondegenerate optical parametric amplifier. The measured distributions exhibit up to 1.9 dB of quantum correlation between the signal and idler photon numbers, whereas the marginal distributions are thermal as expected for parametric fluorescence. PMID- 11018884 TI - Directed current due to broken time-space symmetry AB - We consider the classical dynamics of a particle in a one-dimensional space periodic potential U(X) = U(X+2pi) under the influence of a time-periodic space homogeneous external field E(t) = E(t+T). If E(t) is neither a symmetric function of t nor antisymmetric under time shifts E(t+/-T/2) not equal-E(t), an ensemble of trajectories with zero current at t = 0 yields a nonzero finite current as t- >infinity. We explain this effect using symmetry considerations and perturbation theory. Finally we add dissipation (friction) and demonstrate that the resulting set of attractors keeps the broken symmetry property in the basins of attraction and leads to directed currents as well. PMID- 11018885 TI - How efficiently Do three pointlike particles sample phase space? AB - We show that the continuous phase space of a hard particle system can be mapped onto a discrete but infinite phase space. For three pointlike particles confined to a ring, the evolution of the system maps onto a chaotic walk on a hexagonal lattice. This facilitates direct measurement of the departure of the system from its original configuration. In special cases of mass ratios the phase space becomes closed and finite (nonergodic). There are qualitative differences between this chaotic walk and a random walk, in particular a more rapid sampling of phase space. PMID- 11018886 TI - Estimation of noise levels for models of chaotic dynamical systems AB - We investigate how far it is possible to identify and separate dynamical noise from measurement noise in observed nonlinear time series. Using Bayesian methods, we derive estimates for the two noise levels, and find that, given a good model of the dynamics, these can give accurate results even if the dynamical noise level is orders of magnitude smaller than the measurement noise level, whereas a simple calculation of root mean square error badly understates the dynamical noise. We argue that this allows better estimates of the underlying dynamical time series, and so better predictions of its future and of its fundamental dynamical properties. PMID- 11018887 TI - Average energy flow of optical pulses in dispersive media AB - The arrival time of a light pulse at a point in space is defined using a time expectation integral over the Poynting vector. The delay between pulse arrival times at two distinct points is shown to consist of two parts: a spectral superposition of group delays (inverse of group velocity) and a delay due to spectral reshaping via absorption or amplification. The result provides a context wherein group velocity is always meaningful even for broad band pulses and when the group velocity is superluminal or negative. The result imposes luminality on sharply defined pulses. PMID- 11018888 TI - Bright spatial solitons on a partially incoherent background AB - We present the first observation of incoherent antidark spatial solitons in noninstantaneous nonlinear media. This new class of soliton states involves bright solitons on a partially incoherent background of infinite extent. In the case where the nonlinearity is of the Kerr type, their existence is demonstrated analytically by means of an exact solution. Computer simulations and experiments indicate that these incoherent antidark solitons can propagate in a stable fashion provided that the spatial coherence of their background is reduced below the incoherent modulation instability threshold. PMID- 11018889 TI - Acoustic einstein-hopf drag on a bubble AB - Theoretical results show that the drag on a bubble can be modified by the presence of isotropic, homogeneous, broadband acoustic noise, when the band overlaps the bubble's resonance width. While these results constitute an acoustic analog to the Einstein-Hopf drag on an oscillating dipole in the presence of electromagnetic fluctuations, an important difference is that band-limited acoustic noise can reduce the drag when the lower frequency of the spectrum coincides with the resonant frequency of the bubble. Applications to bubble migration, heat transfer, and acoustophoresis are suggested. PMID- 11018890 TI - Normal heat conductivity of the one-dimensional lattice with periodic potential of nearest-neighbor interaction AB - The process of heat conduction in a chain with a periodic potential of nearest neighbor interaction is investigated by means of molecular dynamics simulation. It is demonstrated that the periodic potential of nearest-neighbor interaction allows one to obtain normal heat conductivity in an isolated one-dimensional chain with conserved momentum. The system exhibits a transition from infinite to normal heat conductivity with the growth of its temperature. The physical reason for normal heat conductivity is the excitation of high-frequency stationary localized rotational modes. These modes absorb the momentum and facilitate locking of the heat flux. PMID- 11018891 TI - Universality and saturation of intermittency in passive scalar turbulence AB - The statistical properties of a scalar field advected by the nonintermittent Navier-Stokes flow arising from a two-dimensional inverse energy cascade are investigated. The universality properties of the scalar field are probed by comparing the results obtained with two types of injection mechanisms. Scaling properties are shown to be universal, even though anisotropies injected at large scales persist down to the smallest scales and local isotropy is not fully restored. Scalar statistics is strongly intermittent and scaling exponents saturate to a constant for sufficiently high orders. This is observed also for the advection by a velocity field rapidly changing in time, pointing to the genericity of the phenomenon. PMID- 11018892 TI - Finite wavelength instabilities in a slow mode coupled complex ginzburg-landau equation AB - In this Letter, we discuss the effect of slow real modes in reaction-diffusion systems close to a supercritical Hopf bifurcation. The spatiotemporal effects of the slow mode cannot be captured by traditional descriptions in terms of a single complex Ginzburg-Landau equation (CGLE). We show that the slow mode coupling to the CGLE introduces a novel set of finite wavelength instabilities not present in the CGLE. For spiral waves, these instabilities highly affect the location of regions for convective and absolute instability. These new instability boundaries are consistent with transitions to spatiotemporal chaos found by simulation of the corresponding coupled amplitude equations. PMID- 11018893 TI - Self-amplification of coherent spontaneous emission in a cherenkov free-electron maser AB - Ultrashort pulses of microwave radiation have been produced in a dielectric-lined Cherenkov free-electron maser (FEM) amplifier. An intense initial seed pulse, due to coherent spontaneous emission (CSE), arises at the leading edge of the electron pulse. There is evidence to show that 3-4 cycle spikes are produced through the amplification of these seed pulses. A strong dependence of the start up power on the rise time of the electron pulse has been found. The experimental results are verified by a theoretical analysis. Our study shows that amplification in a FEM amplifier is always initiated by CSE arising from the edge of the electron pulse when the rise time is comparable to the electromagnetic wave period. PMID- 11018894 TI - Identification of minority ion-cyclotron emission during radio frequency heating in the JET tokamak AB - First measurements and identification of minority ion-cyclotron emission (MICE) during ICRF (H)D minority heating in the JET tokamak are presented. An inner wall radiofrequency (rf) probe shows the new single MICE spectral line, down-shifted from the heating frequency and appearing approximately 400 ms after the ICRH switch-on. The line is narrow (Deltaomega/omega approximately 0.04), characterized by the ion-cyclotron frequency of minority protons in the outer edge midplane plasma and is observed irrespective of whether single or multifrequency ICRH is applied. The observations are consistent with the classical evolution and population of the plasma edge with approximately 3 MeV ICRH protons on orbits near the outboard limiters. Particle loss and energy filtering contribute to a local non-Maxwellian energetic ion distribution, which is susceptible to ion-cyclotron instability. PMID- 11018895 TI - Destabilization of the m = 1 diocotron mode in non-neutral plasmas AB - The theory for a Penning-Malmberg trap predicts m = 1 diocotron stability. However, experiments with hollow profiles show robust exponential growth. We propose a new mechanism of destabilization of this mode, involving parallel compression due to end curvature. The results are in good agreement with the experiments. The resulting modified drift-Poisson equations are analogous to the geophysical shallow water equations, and conservation of line integrated density corresponds to that of potential vorticity. This analogy predicts Rossby waves in non-neutral plasmas and an m = 1 instability in fluids. PMID- 11018896 TI - Laser-hole boring into overdense plasmas measured with soft X-Ray laser probing AB - A laser self-focused channel formation into overdense plasmas was observed using a soft x-ray laser probe system with a grid image refractometry (GIR) technique. 1.053 &mgr;m laser light with a 100 ps pulse duration was focused onto a preformed plasma at an intensity of 2x10(17) W/cm (2). Cross sections of the channel were obtained which show a 30 &mgr;m diameter in overdense plasmas. The channel width in the overdense region was kept narrow as a result of self focusing. Conically diverging density ridges were also observed along the channel, indicating a Mach cone created by a shock wave due to the supersonic propagation of the channel front. PMID- 11018897 TI - Devil's staircase-type faceting of a cubic lyotropic liquid crystal AB - The faceting of monocrystals of the lyotropic cubic liquid crystals in equilibrium with a humid atmosphere is observed. Experiments reveal the presence of more than 60 different types of facets on the surface of a spherical crystal of radius R = 1 mm. The devil's staircase type of faceting has been predicted theoretically when the interaction between steps on the crystal surface is repulsive. PMID- 11018898 TI - Defects in a disordered world: the structure of glassy GeSe2 AB - The full set of partial structure factors for the prototypical network glass GeSe2 was measured using the method of isotopic substitution in neutron diffraction. The basic building block of the network is the Ge(Se(1/2))(4) tetrahedron in which 34(5)% of the Ge reside in edge-sharing configurations. The intrinsic chemical order of the glass is, however, broken with a maximum of 25(5)% Ge and 20(5)% Se being involved in homopolar bonds at distances of 2.42(2) and 2.32(2) A, respectively. PMID- 11018899 TI - Unexpected scenario of glass transition in polymer globules: an exactly enumerable model. AB - We introduce a lattice model of glass transition in polymer globules. This model exhibits ergodicity breaking in which the disjoint regions of phase space do not arise uniformly, but as small chambers whose number increases exponentially with polymer density. Chamber sizes obey power law distribution, making phase space similar to a fractal foam. This clearly demonstrates the importance of the phase space geometry and topology in describing any glass-forming system, such as semicompact polymers during protein folding. PMID- 11018900 TI - Observation of "Stick" and "Handle" intermediates along the fullerene road AB - The hypothesis that fullerenes grow in a carbon plasma by the addition of C2 units (the "fullerene road") has been widely acclaimed as the most plausible mechanism for formation of larger fullerenes including C60 and C70. Calculations suggest that the association of C2 with fullerenes proceeds through two classes of intermediates, "sticks" and "handles." Here we report the observation of these species using high-resolution ion-mobility measurements for C(n) cations generated by laser vaporization of graphite and laser desorption of C60. Sticks with up to eight-atom chains have also been found. PMID- 11018901 TI - Dimer to monomer phase transition in alkali-metal fullerides: magnetic susceptibility changes AB - Ab initio calculations have been employed to investigate the peculiar change in magnetic property (from diamagnetic to paramagnetic) of the dianionic C60-dimer phase in a rapidly cooled AC60 samples ( A: alkali metal). We first note that the triplet state of (C60)-22 which was never considered previously is nearly degenerate with the singlet state, and the transition barrier between the two states is reasonably small. This explains the susceptibility increase with an increase in temperature and the magnetic phase transition in the process of the dimer to monomer phase transition. PMID- 11018902 TI - Water under pressure AB - Water under pressure is investigated by first principles molecular dynamics, with a focus on the changes in hydrogen bonding and the oxygen network in the nondissociative regime. At a pressure of 10 GPa and a temperature of 600 K, which is close to the freezing point, no appreciable molecular dissociation is observed in the simulations. However, the structure of water is substantially altered from that at ambient conditions. The liquid exhibits a much larger coordination of oxygen atoms, an essential weakening of hydrogen bonding, and sizable changes in the density of electronic states close to the Fermi level. Our results provide new structural data for direct comparison with future experiments. PMID- 11018903 TI - Metastability and nucleation in capillary condensation AB - This paper is devoted to thermally activated dynamics of capillary condensation. On the basis of a simple model we identify the critical nucleus involved in the transition mechanism and calculate the nucleation barrier from which we obtain information on the nucleation time. Close to the condensation point, the theory predicts extremely large energy barriers leading to strong metastabilities, long time dependencies, and large hysteresis in agreement with experimental observations in mesoporous media. The validity of the model is assessed using a numerical simulation of a time-dependent Ginzburg-Landau model for the confined system. PMID- 11018904 TI - Orientational dynamics in supercooled liquids near T(c) and comparison with ideal mode-coupling theory AB - Orientational dynamics in supercooled salol and ortho-terphenyl were measured near their critical temperatures, T(c), with optical Kerr effect experiments spanning a very broad range of times. Above T(c), the decays are shown to be in excellent agreement with the master curve predicted by ideal mode-coupling theory when higher order terms are included. Between the critical decay and the von Schweidler power laws, the intermediate time range of the data can be modeled by a power law. This intermediate power law, located at 2 which is the thermodynamically favored orientation. As the deposition temperature is decreased to 130 K, an unexpected 90 degrees switch of the ripple orientation is observed. The ridges of the ripples and the steps are in this case parallel to <100>. In the intermediate temperature range a checkerboard of rectangular mounds results. We interpret our results in terms of the peculiar hierarchy of interlayer and intralayer diffusion barriers present on the anisotropic Ag(110) surface. PMID- 11018907 TI - Enhanced instability of strained alloy films due to compositional stresses AB - A single-component strained film is known to be unstable to the stress-driven morphological instability. Here, we determine how the instability is modified in an alloy film by considering the effect of compositional stresses due to an atomic size difference. We find that the coupling of composition to stress always makes the film more unstable to the formation of stress-driven surface undulations. The destabilization is greatest over a range of intermediate deposition rates. PMID- 11018908 TI - Gapping by squashing: metal-insulator and insulator-metal transitions in collapsed carbon nanotubes AB - Squashing brings circumferentially separated areas of a carbon nanotube into close proximity, drastically altering the low-energy electronic properties and (in some cases) reversing standard rules for metallic versus semiconducting behavior. Such a deformation mode, not requiring motion of tube ends, may be useful for devices. Uniaxial stress of a few kbar can reversibly collapse a small radius tube, inducing a 0.1 eV gap with a very strong pressure dependence, while the collapsed state of a larger tube is stable. The low-energy electronic properties of chiral tubes are surprisingly insensitive to collapse. PMID- 11018909 TI - Excitonic and quasiparticle gaps in Si nanocrystals AB - We present calculations of the one- and two-particle excitations in silicon nanocrystals. The one-particle properties are handled in the GW approximation, and the excitonic gap is obtained from the Bethe-Salpeter equation. We develop a tight binding version of these methods to treat clusters up to 275 atoms. The self-energy and Coulomb corrections almost exactly cancel each other for crystallites with radius larger than 0.6 nm. The result of this cancellation is that one-particle calculations give quite accurate values for the excitonic gap of crystallites in the most studied range of sizes. PMID- 11018910 TI - Competition between phase separation and "Classical" intermediate valence in an exactly solved model AB - The exact solution of the spin-1 / 2 Falicov-Kimball model on an infinite coordination Bethe lattice is analyzed in the regime of "classical" intermediate valence. We find that (i) either phase separation or a direct metal-insulator transition precludes intermediate valence over a large portion of the phase diagram, and (ii) within the intermediate valence phase, only continuous transitions are found as functions of the localized f-electron energy or temperature. PMID- 11018911 TI - Nonlinear electric field effects at a continuous mott-hubbard transition AB - We characterize the non-Ohmic portion of the conductivity at temperatures T<1 K in the highly correlated transition metal chalcogenide Ni(S,Se)(2). Pressure tuning of the T = 0 metal-insulator transition reveals the influence of the quantum critical point and permits a direct determination of the dynamical critical exponent z = 2.7(+0.3)(-0.4). Within the framework of finite temperature scaling, we find that the spatial correlation length exponent nu and the conductivity exponent &mgr; differ. PMID- 11018912 TI - Dynamical properties near the metal-insulator transition: evidence for electron assisted variable range hopping AB - We report independent measurements (between 20 and 200 mK) of the electronic specific heat C(e), the electron-phonon coupling G(e-ph), and the electron-phonon relaxation time tau(e-ph) (from 10(-2) to 10(-5) s) for NbxSi1-x Anderson insulator thin films. We show that the usual equation tau(e-ph) = C(e)/G(e-ph) holds only if the resistance is solely related to the electron temperature. We conclude that at sufficiently low temperatures variable range hopping transport is assisted by electron-electron interactions alone and is independent of the phonon distribution. PMID- 11018913 TI - Probing the potential landscape inside a two-dimensional electron Gas AB - We report direct observations of the scattering potentials in a two-dimensional electron gas using electron-beam diffraction experiments. The diffracting objects are local density fluctuations caused by spatial and charge-state distribution of donors in the GaAs-(Al,Ga)As heterostructures. The scatterers can be manipulated externally by sample illumination or by cooling the sample down under depleted conditions. PMID- 11018914 TI - Topological scenario for stripe formation in manganese oxides AB - The spin-charge-orbital complex structures of manganites are studied using topological concepts. The key quantity is the "winding number" w associated with the Berry-phase connection of an e(g) electron parallel transported through Jahn Teller centers, along zigzag one-dimensional paths in an antiferromagnetic environment of t(2g) spins. From these concepts, it is shown that the "bi-stripe" and "Wigner-crystal" states observed experimentally have different w's. Predictions for the spin structure of the charge-ordered states for heavily doped manganites are discussed. PMID- 11018915 TI - Finite-element theory of transport in ferromagnet-normal metal systems AB - We formulate a theory of spin dependent transport in an electronic circuit involving ferromagnetic elements with noncollinear magnetization which is based on the conservation of spin and charge current. The theory considerably simplifies the calculation of the transport properties of complicated ferromagnet normal metal systems. We illustrate the theory by considering a novel three terminal device. PMID- 11018916 TI - Direct coulomb and exchange interaction in artificial atoms AB - We determine contributions from the direct Coulomb and exchange interactions to the total interaction in artificial semiconductor atoms. We tune the relative strengths of the two interactions and measure them as a function of the number of confined electrons. The electrons tend to have parallel spins when they occupy nearly degenerate single-particle states. We use a magnetic field to adjust the single-particle-state degeneracy, and find that the spin configurations in an arbitrary magnetic field are well explained in terms of two-electron singlet and triplet states. PMID- 11018917 TI - Weak localization, hole-hole interactions, and the "Metal"-insulator transition in two dimensions AB - A detailed investigation of the metallic behavior in high-quality GaAs-AlGaAs two dimensional hole systems reveals the presence of quantum corrections to the resistivity at low temperatures. Despite the low density ( r(s)>10) and high quality of these systems, both weak localization (observed via negative magnetoresistance) and weak hole-hole interactions (giving a correction to the Hall constant) are present in the so-called metallic phase where the resistivity decreases with decreasing temperature. If these quantum corrections persist down to T = 0, the results suggest that even at high r(s) there is no metallic phase in two dimensions. PMID- 11018918 TI - Dynamic phases and the peak effect in dirty type II superconductors AB - We study numerically and experimentally the dynamics of driven vortex matter. Our London-Langevin simulations find that the critical current exhibits a peak both across the Bragg glass to vortex glass transition and across the melting line. The peak is accompanied by a clear crossing of the I-V curves. We report transport measurements on untwinned YBCO crystals, in complete accordance with these findings. At higher drives disorder is averaged to reduced values, and in three dimensions the vortices reorder into a "moving solid." The effect of the disorder can be well represented with a "shaking temperature" which is inversely proportional to the velocity. PMID- 11018919 TI - Enhanced critical currents of superconducting ErNi2B2C in the ferromagnetically ordered state AB - We report on transport and magnetization studies of the critical current in single crystal ErNi2B2C for applied fields below 4 kG. Below T approximately 2.5 K superconductivity coexists with weak ferromagnetism. We find that the critical currents are strongly enhanced for all field orientations in this ferromagnetic regime, corresponding to a threefold increase of the pinning force of the flux line lattice. We speculate that this increase is due to strong pair breaking by the ferromagnetism. PMID- 11018920 TI - Magnetoresistance of ferromagnetic tunnel junctions in the double-exchange model AB - We conduct a theoretical study of the temperature dependence of the spin polarization ( P) and the magnetoresistance (MR) ratio using the double exchange (DE) model for ferromagnetic tunnel junctions with half-metallic systems. It is shown that the strong exchange coupling in the DE model plays an important role in the temperature dependence of both P and the MR ratio; their values can be less than the maximum values expected for half-metallic systems at low temperatures, and the MR ratio decreases more rapidly than P with increasing temperature. The calculated results, however, indicate that the MR ratio may still be large at high temperatures near the Curie temperature. PMID- 11018921 TI - Optical anisotropy of zinc-blende semiconductors in an electric field AB - A theory of optical anisotropy for zinc-blende semiconductors in an electric field is derived by extending the formalism of Luttinger and Kohn [Phys. Rev. 97, 869 (1955)] to higher order. This resolves a recent controversy over the correct form of the Hamiltonian for the degenerate valence bands. PMID- 11018922 TI - Squeezing alcohols into sonoluminescing bubbles: the universal role of surfactants AB - We conduct an experimental study of the dependence of single bubble sonoluminescence intensity on the concentration of various alcohols. The light intensity is reduced by one-half at a molar fraction of ethanol of approximately 2.5x10(-5); butanol achieves the same reduction at a concentration 10 times smaller. We account for the results by a theoretical model in which the alcohols are assumed to be mechanically forced into the bubble at collapse, modifying the adiabatic exponent of the gas. The increasing hydrophobicities of the alcohols lead to decreasing effective adiabatic exponents, and thus to less heating and therefore less light. Support for this model is obtained by replotting the experimental light intensity values vs the calculated exponents, yielding a collapse of all data onto a universal curve. PMID- 11018923 TI - Regulated and entangled photons from a single quantum Dot AB - We propose a new method of generating nonclassical optical field states. The method uses a semiconductor device, which consists of a single quantum dot as active medium embedded in a p- i- n junction and surrounded by a microcavity. Resonant tunneling of electrons and holes into the quantum dot ground states, together with the Pauli exclusion principle, produce regulated single photons or regulated pairs of photons. We propose that this device also has the unique potential to generate pairs of entangled photons at a well-defined repetition rate. PMID- 11018924 TI - Role of the 2D surface state continuum and projected band gap in charge transfer in front of a Cu(111) surface AB - Electron capture by Li+ and H projectiles in grazing scattering from Cu(111) and Cu(110) surfaces is studied experimentally and theoretically. Whereas data for Cu(110) can be described by established theoretical methods treating resonant charge transfer with a free-electron metal, data for Cu(111) show pronounced deviations from this approach. We interpret our observations by the effect of the projected L-band gap of the Cu(111) surface. In particular, the quantum states of reduced dimension (2D surface state continuum) play a dominant role in electron transfer. PMID- 11018925 TI - Classical teleportation of a quantum Bit AB - Classical teleportation is defined as a scenario where the sender is given the classical description of an arbitrary quantum state while the receiver simulates any measurement on it. This scenario is shown to be achievable by transmitting only a few classical bits if the sender and receiver initially share local hidden variables. Specifically, a communication of 2.19 bits is sufficient on average for the classical teleportation of a qubit, when restricted to von Neumann measurements. The generalization to positive-operator-valued measurements is also discussed. PMID- 11018926 TI - Theory of quantum error correction for general noise AB - A measure of quality of an error-correcting code is the maximum number of errors that it is able to correct. We show that a suitable notion of "number of errors" e makes sense for any quantum or classical system in the presence of arbitrary interactions. Thus, e-error-correcting codes protect information without requiring the usual assumptions of independence. We prove the existence of large codes for both quantum and classical information. By viewing error-correcting codes as subsystems, we relate codes to irreducible representations of operator algebras and show that noiseless subsystems are infinite-distance error correcting codes. PMID- 11018927 TI - Highly optimized tolerance: robustness and design in complex systems AB - Highly optimized tolerance (HOT) is a mechanism that relates evolving structure to power laws in interconnected systems. HOT systems arise where design and evolution create complex systems sharing common features, including (1) high efficiency, performance, and robustness to designed-for uncertainties, (2) hypersensitivity to design flaws and unanticipated perturbations, (3) nongeneric, specialized, structured configurations, and (4) power laws. We study the impact of incorporating increasing levels of design and find that even small amounts of design lead to HOT states in percolation. PMID- 11018928 TI - Do free DNA counterions control the osmotic pressure? AB - The contribution of counterions to macroscopic properties of isotropic DNA solutions has been studied using osmotic pressure measurements in low added salt condition. In the high DNA concentration range, the counterion contribution prevails and the associated osmotic coefficient is equal to 0.245+/-0.020. In the lower concentration range, the osmotic pressure may be exerted either by polymers or by ions, or due to a combination of both effects, depending on the added salt and DNA concentrations. PMID- 11018929 TI - Twist in chiral interaction between biological helices. AB - Using an exact solution for the pair interaction potential, we show that long, rigid, chiral molecules with helical surface charge patterns have a preferential interaxial angle approximately sqrt[RH]/L, where L is the length of the molecules, R is the closest distance between their axes, and H is the helical pitch. Estimates based on this formula suggest a solution for the puzzle of small interaxial angles in alpha-helix bundles and in cholesteric phases of DNA. PMID- 11018930 TI - Comment on "Accelerated emission of gamma rays from the 31-yr isomer of 178Hf induced by X-Ray Irradiation" PMID- 11018931 TI - Comment on "Accelerated emission of gamma rays from the 31-yr isomer of 178Hf induced by X-Ray Irradiation" PMID- 11018932 TI - Comment on "Accelerated emission of gamma rays from the 31-yr isomer of 178Hf induced by X-Ray Irradiation" PMID- 11018934 TI - Comment on "Singularities and pseudogaps in the density of states of peierls chains" PMID- 11018933 TI - Collins et al. reply: PMID- 11018935 TI - Bartosch and kopietz reply: PMID- 11018936 TI - Comment on "Density functional simulation of a breaking nanowire" PMID- 11018937 TI - Nakamura et al. reply: PMID- 11018938 TI - Incompatibility of macroscopic local realism with quantum mechanics in measurements with macroscopic uncertainties AB - We show that quantum mechanics predicts a contradiction with local hidden variable theories for photon number measurements which have limited resolving power, to the point of imposing an uncertainty in the photon number result which is macroscopic in absolute terms. We show how this can be interpreted as a failure of a new premise, macroscopic local realism. PMID- 11018939 TI - Interpretation of the nonextensivity parameter q in some applications of tsallis statistics and Levy distributions AB - The nonextensivity parameter q occurring in some of the applications of Tsallis statistics (known also as index of the corresponding Levy distribution) is shown to be given, in the q>1 case, entirely by the fluctuations of the parameters of the usual exponential distribution. PMID- 11018940 TI - Topology and phase transitions: paradigmatic evidence AB - We report upon the numerical computation of the Euler characteristic chi (a topologic invariant) of the equipotential hypersurfaces Sigma(v) of the configuration space of the two-dimensional lattice varphi(4) model. The pattern chi(Sigma(v)) versus v (potential energy) reveals that a major topology change in the family Sigma(v)(vinR) is at the origin of the phase transition in the model considered. The direct evidence given here-of the relevance of topology for phase transitions-is obtained through a general method that can be applied to any other model. PMID- 11018941 TI - Gravity in the randall-sundrum brane world AB - We discuss the weak gravitational field created by isolated matter sources in the Randall-Sundrum brane world. For the case of a single wall of positive tension, the field stays localized near the wall if the source is stationary. We calculate the leading Kaluza-Klein corrections to the linearized gravitational field of a nonrelativistic spherical object, which is different from the Schwarzschild solution at large distances. In the case of two branes of opposite tension, linearized Brans-Dicke (BD) gravity is recovered on either wall, with different BD parameters. On the wall with positive tension the BD parameter is larger than 3000 provided that the separation between walls is larger than 4 times the AdS radius. The gravitational field due to shadow matter is also considered. PMID- 11018942 TI - The pinch technique at two loops AB - It is shown that the fundamental properties of gauge independence, gauge invariance, unitarity, and analyticity of the S matrix lead to the unambiguous generalization of the pinch technique algorithm to two loops. PMID- 11018943 TI - Isolated photon cross section in p&pmacr; collisions at radicals = 1. 8 TeV AB - We report a new measurement of the cross section for the production of isolated photons with transverse energies ( E(gamma)(T)) above 10 GeV and pseudorapidities |eta|<2.5 in p&pmacr; collisions at sqrt[s] = 1.8 TeV. The results are based on a data sample of 107.6 pb(-1) recorded during 1992-1995 with the D0 detector at the Fermilab Tevatron collider. The background, predominantly from jets which fragment to neutral mesons, was estimated using the longitudinal shower shape of photon candidates in the calorimeter. The measured cross section is in good agreement with the next-to-leading order QCD calculation for E(gamma)(T) greater, similar36 GeV. PMID- 11018944 TI - Differential production cross section of Z bosons as a function of transverse momentum at sqrt AB - We present a measurement of the transverse momentum distribution of Z bosons produced in p&pmacr; collisions at sqrt[s] = 1.8 TeV from data collected by the DO experiment at the Fermilab Tevatron Collider. We find good agreement between our results and current resummation calculations, and also use our data to extract nonperturbative parameters for a particular version of the resummation formalism. The resulting values are significantly more precise than obtained in previous determinations. PMID- 11018945 TI - Bombarding energy dependence of pi(-) interferometry at the brookhaven AGS AB - We present the first excitation function of pi(-) intensity interferometry at Alternating Gradient Synchrotron (AGS) energies (2-8 A GeV). The sensitivity of the multidimensional correlation functions to the geometry and dynamics of the pion-emitting system provides a stringent test of transport models of heavy ion collisions. Detailed comparisons with a realistic transport model, both with and without an explicit nuclear mean field, suggest that the beam energy evolution in the reaction dynamics is different in the model than in the data. A significantly increased pi(-) emission time scale, which has been suggested as a signal of the onset of the transition to quark-gluon plasma, is not observed. PMID- 11018946 TI - Novel rapidity dependence of directed flow in high-energy heavy-Ion collisions AB - For high-energy nucleus-nucleus collisions, we show that a combination of space momentum correlations characteristic of radial expansion, together with the correlation between the position of a nucleon in the nucleus and its stopping, results in a very specific rapidity dependence of directed flow: a reversal of sign in the midrapidity region. We support our argument by RQMD model calculations for Au+Au collisions at sqrt[s] = 200A GeV. PMID- 11018947 TI - Enhanced cooling of hydrogen atoms by lithium atoms AB - We present calculated scattering lengths for collisions between various isotopic forms of lithium and hydrogen atoms interacting via singlet and triplet molecular states of LiH. We demonstrate that one bound triplet level is supported for each isotopomer 7LiH, 6LiH, 7LiD, and 6LiD. We obtain large calculated triplet scattering lengths that are stable against uncertainties in the potential. We present elastic and momentum transfer cross sections, and the corresponding rate coefficients, for hydrogen atoms colliding with 7Li atoms. We suggest that enhanced cooling of trapped atomic hydrogen by 7Li atoms is feasible. PMID- 11018948 TI - Autoionization of triply excited rydberg series AB - The Auger rates of triply excited Rydberg series are shown to behave rather differently from doubly excited series. It is shown that in hollow atoms the Auger decay rates for Rydberg series of the type 2l2l(')nl(") with n>/=2 are expected to be nearly independent of n, while for doubly excited series of the type 2lnl(') the decay rate in general decreases with increasing n. In addition the ratio between the rates for 2l2l(')nl(") Rydberg series with different l(") values will be fixed and often the ratio will be equal to one. PMID- 11018949 TI - Cold rubidium molecules formed in a magneto-optical trap AB - We report the first observation of translationally cold ( approximately 90 &mgr;K) Rb2 molecules. They are produced in a magneto-optical trap in their triplet ground state. The detection is performed by selective mass spectroscopy after two-photon ionization into Rb+2, resonantly enhanced through the intermediate a (3)Sigma(+)(u)-->2 (3)Pi(g) molecular band. The two rubidium isotopes present very different types of behavior that are interpreted in terms of their respective collisional properties. PMID- 11018950 TI - Quantum entanglement via optical control of atom-atom interactions AB - Two-photon optical transitions combined with long-range dipole-dipole interactions can be used for the coherent manipulation of multiatom collective states. We show that it is possible to induce optical resonances accompanied by the generation of entangled superpositions of such atomic states. Resonances of this kind can be used to implement quantum logic gates using optically excited single atoms (impurities) in the condensed phase. PMID- 11018951 TI - Strong species dependence of high order photoelectron production in alkali metal atoms AB - We present a theoretical and experimental study of the production of very high order photoelectrons from alkali metal atoms interacting with intense, midinfrared radiation. The strength of this process shows an unexpectedly strong species dependence. We find that this dependence can be explained via the difference in the cross section for electron-ion scattering from the different atoms. This allows us to directly relate the high energy portion of the photoelectron spectrum to specific features of the electron-ion potential. PMID- 11018952 TI - Doppler splitting of In-flight auger decay of dissociating oxygen molecules: the localization of delocalized core holes AB - By exploiting the core-excitation-induced dissociation of O2, we find that the Auger emission exhibits a Doppler-like energy shift. We show this to be a manifestation of localization of the core hole and propose that the problem of core-hole localization versus delocalization in core-hole spectroscopies may be resolved by considering the nature of the measurement. PMID- 11018953 TI - Photoinduced polarization inversion in a polymeric molecule AB - The polymeric molecule can exhibit a new photoinduced phenomenon where the electric dipole of the molecule with a bipolaron is reversed by absorbing one photon. This photoinduced polarization inversion occurred in a single molecule is an ultrafast process with a relaxation time of 200 fs. PMID- 11018954 TI - Ergodicity of random-matrix theories: the unitary case AB - We prove ergodicity of unitary random-matrix theories by showing that the autocorrelation function with respect to energy or magnetic field strength of any observable vanishes asymptotically. We do so using Efetov's supersymmetry method, a polar decomposition of the saddle-point manifold, and an asymptotic evaluation of the boundary terms generated in this fashion. PMID- 11018955 TI - Relaxation and diffusion for the kicked rotor AB - The dynamics of the kicked rotor, which is a paradigm for a mixed system, where the motion in some parts of phase space is chaotic and in other parts is regular, is studied statistically. The evolution operator of phase space densities in the chaotic component is calculated in the presence of noise, and the limit of vanishing noise is taken in the end. The relaxation rates to the equilibrium density are calculated analytically within an approximation that improves with increasing stochasticity. The results are tested numerically. A global picture is presented of relaxation to the equilibrium density in the chaotic component when the system is bounded and to diffusive behavior when it is unbounded. PMID- 11018956 TI - Unification of perturbation theory, random matrix theory, and semiclassical considerations in the study of parametrically dependent eigenstates AB - We consider a classically chaotic system that is described by a Hamiltonian H(Q,P;x), where x is a constant parameter. Specifically, we discuss a gas particle inside a cavity, where x controls a deformation of the boundary or the position of a "piston." The quantum eigenstates of the system are |n(x)>. We describe how the parametric kernel P(nmid R:m) = ||(2) evolves as a function of deltax = x-x(0). We explore both the perturbative and the nonperturbative regimes, and discuss the capabilities and the limitations of semiclassical as well as random waves and random-matrix-theory considerations. PMID- 11018957 TI - Brewster's scattering angle in scattered waves from slightly rough metal surfaces AB - Brewster's scattering angle in electromagnetic wave scattering from slightly random metal surfaces is investigated by means of the stochastic functional approach. While there are dips due to Brewster's scattering angle in scattering profiles from dielectric surfaces, Brewster's scattering angle does not exist in scattering from metal surfaces. However, the dips can exist in scattering from rough metal surfaces with the optically denser medium to convert evanescent wave into radiative wave. PMID- 11018958 TI - Observation of a cw dark-field signal in an absorption spectroscopic experiment using a phase locked free-electron laser AB - We introduce a technique for ultrasensitive absorption spectroscopy using the GHz rate pulse train from a phase-locked free-electron laser (FEL), in which the fractional power absorbed from one or more laser lines reappears as a signal on the dark background between the pulses emerging from the sample. Preliminary absorption experiments in 15 Torr cm of methane at 3.25 &mgr;m, using phase locked pulses from the Mark III FEL, clearly reveal an interpulse beat signal due to absorption by adjacent molecular rotational lines which is generated only in the presence of interpulse phase coherence. PMID- 11018959 TI - Robust photonic band gap from tunable scatterers AB - We show theoretically and experimentally that photonic band gaps can be realized using metal or metal-coated spheres as building blocks. Robust photonic gaps exist in any periodic structure built from such spheres when the filling ratio of the spheres exceeds a threshold. The frequency and the size of the gaps depend on the local order rather than on the symmetry or the global long range order. Good agreement between theory and experiment is obtained in the microwave regime. Calculations show that the approach can be scaled up to optical frequencies even in the presence of absorption. PMID- 11018960 TI - Momentum conservation implies anomalous energy transport in 1D classical lattices AB - Under quite general conditions, we prove that for classical many-body lattice Hamiltonians in one dimension (1D) total momentum conservation implies anomalous conductivity in the sense of the divergence of the Kubo expression for the coefficient of thermal conductivity, kappa. Our results provide rigorous confirmation and explanation of many of the existing "surprising" numerical studies of anomalous conductivity in 1D classical lattices, including the celebrated Fermi-Pasta-Ulam problem. PMID- 11018961 TI - Two-color operation in high-gain free-electron lasers AB - Two-color operation in free-electron laser (FEL) amplifiers is studied using a 3D nonlinear polychromatic simulation. We assume the FEL is seeded at two closely spaced wavelengths within the gain band, and study the growth of the seeds and a discrete spectrum of beat waves that are outside the gain band. The beat waves grow parasitically due to electron bunching in the seeded waves with growth rates higher than the seeded waves. Injection of narrow-band seeds ensures a discrete spectrum. An example is discussed corresponding to an x-ray FEL; however, the physics is applicable to all spectral ranges. PMID- 11018962 TI - New Alfven continuum gaps and global modes induced by toroidal flow AB - Continuous magnetohydrodynamic (MHD) spectra in tokamaks with toroidal rotation are studied. In the corotating frame, the linear perturbations experience Coriolis and centrifugal effects. The latter create Alfven frequency gaps on rational surfaces resonant with the perturbations. New global eigenmodes are located inside these gaps. These modes may be used for MHD spectroscopy. In contrast to the usual gaps and toroidal Alfven eigenmodes, the new flow-induced gaps and global modes are in the low frequency range so that they will have important implications for stability as well. PMID- 11018963 TI - First observation of ion acoustic waves produced by the langmuir decay instability AB - Thomson scattering measurements are presented which demonstrate conclusively the occurrence of the Langmuir decay instability (LDI) in a laser-produced plasma experiment. Both products of the instability, the ion acoustic wave and the electron plasma wave, were simultaneously observed and identified with their spectral characteristics. The secondary decay of the LDI-generated electron plasma wave, into another Langmuir wave and an ion acoustic wave, has been observed for the first time. The connection with growth and saturation of the stimulated Raman instability is discussed. PMID- 11018964 TI - Dynamics of fast electrons driven by the lower hybrid wave in giant sawtoothing plasmas AB - By hard x-ray tomography of the nonthermal bremsstrahlung emission, a detailed investigation of the dynamics of the fast electrons driven by the lower hybrid wave in the presence of giant sawteeth is carried out. While sawtooth activity is clearly observed on the bulk contribution, no modulation is detected on radiations emitted by the suprathermal population. This original effect is interpreted as the consequence of a poor penetration of the wave in the core of the plasma due to reduced accessibility conditions. Implications for current drive in sawtoothing plasmas are discussed. PMID- 11018965 TI - Correlations between the scanning tunneling microscopy imaged configurations and the electronic structure on stepped Si(111)-( 7x7) surfaces AB - We have observed the dependence of the scanning tunneling microscopy (STM) imaged atom intensity within the (7x7) unit cell on stepped Si(111) as a function of the tunneling voltage. Pronounced differences from the corresponding atom intensity on the flat surface are observed for the contrast of atoms on the low versus the high side of the step and for the contrast between the faulted versus unfaulted subcells of the (7x7) structure. These differences can be accounted for by changes in the electronic structure within the (7x7) subcells adjacent to the step. Calculations of the local density of states and the STM images using a tight-binding method are in excellent agreement with the experimental results. PMID- 11018966 TI - Structures of high-density and low-density water AB - The three site-site partial structure factors for water have been measured as a function of pressure, using neutron diffraction, at a temperature of 268 K. It is found that the measured structure functions imply a continuous transformation with increasing pressure from a low-density form of water ( rho(L) approximately 0.0295 molecules/A(3)), with an open, hydrogen-bonded tetrahedral structure, to a high-density form of water ( rho(H) approximately 0.0402 molecules/A(3)), with nontetrahedral O-O-O angles and a collapsed second coordination shell, which implies broken hydrogen bonds between the first and second coordination shells. PMID- 11018967 TI - Compositional patterning in systems driven by competing dynamics Of different length scale AB - We study an alloy system where short-ranged, thermally driven diffusion competes with externally imposed, finite-ranged, athermal atomic exchanges, as is the case in alloys under irradiation. Using a Cahn-Hilliard-type approach, we show that when the range of these exchanges exceeds a critical value labyrinthine concentration patterns at a mesoscopic scale can be stabilized. Furthermore, these steady-state patterns appear only for a window of the frequency of forced exchanges. Our results suggest that ion beams may provide a novel route to stabilize and tune the size of nanoscale structural features in materials. PMID- 11018968 TI - Nanocrystalline versus microcrystalline Li(2)O:B(2)O3 composites: anomalous ionic conductivities and percolation theory AB - We study ionic transport in nano- and microcrystalline (1-x)Li(2)O:xB(2)O3 composites using standard impedance spectroscopy. In the nanocrystalline samples (average grain size of about 20 nm), the ionic conductivity sigma(dc) increases with increasing content x of B2O3 up to a maximum at x approximately 0.5. Above x approximately 0.92, sigma(dc) vanishes. By contrast, in the microcrystalline samples (grain size about 10 &mgr;m), sigma(dc) decreases monotonically with x and vanishes above x approximately 0. 55. We can explain this strikingly different behavior by a percolation model that assumes an enhanced conductivity at the interfaces between insulating and conducting phases in both materials and explicitly takes into account the different grain sizes. PMID- 11018969 TI - Molecular traffic control in a nanoscale system AB - Atomistic molecular dynamics simulations are used to elucidate a novel and exploitable transport phenomenon known as "molecular traffic control." Under some conditions a binary mixture of differently sized molecules in a structure possessing dual sized pores can exhibit a surprising effect. In the case examined, size segregation and other effects lead to physical separation of the two species through anisotropic diffusion. We have established the underlying causes of this effect in an equilibrium system and used simulations of a relaxing system to show that these causes also hold under nonequilibrium conditions. PMID- 11018970 TI - Atomic structure of the polar NiO(111)- p(2x2) surface AB - Using grazing-incidence x-ray diffraction, the p(2x2) surface structures of the single crystal NiO(111) and a 5 monolayer thick NiO(111) film on Au(111) were both shown to exhibit locally the theoretically predicted octopolar reconstruction, with some important differences. The single crystal exhibits a single Ni termination with double steps. The thin film exhibits both possible terminations (O and Ni) and single steps. These surfaces were found to be nonreactive with respect to hydroxylation. PMID- 11018971 TI - Microstructure controlled shear band pattern formation and enhanced plasticity of bulk metallic glasses containing in situ formed ductile phase dendrite dispersions AB - Results are presented for a ductile metal reinforced bulk metallic glass matrix composite based on glass forming compositions in the Zr-Ti-Cu-Ni-Be system. Primary dendrite growth and solute partitioning in the molten state yields a microstructure consisting of a ductile crystalline Ti-Zr-Nb beta phase, with bcc structure, in a Zr-Ti-Nb-Cu-Ni-Be bulk metallic glass matrix. Under unconstrained mechanical loading organized shear band patterns develop throughout the sample. This results in a dramatic increase in the plastic strain to failure, impact resistance, and toughness of the metallic glass. PMID- 11018972 TI - A nearly universal critical conductivity for semiconductor-metal alloys AB - Classical ionized impurity scattering is employed to calculate the conductivity at and in the vicinity of the critical point. The result sigma(iis)(x = x(c),T) = Asqrt[T], closely given by e(2)/Planck's over 2pilambda(dB) with the de Broglie wavelength lambda(dB) = h/(2m(*)kT)(1/2) in the nondegenerate regime epsilon(F)<x(c), T) might also explain the linear scaling behavior sigma(x, T)-Asqrt[T] = sigma(0)(x/x(0)-1). PMID- 11018973 TI - Metallic low-temperature resistivity in a 2D electron system over an extended temperature range AB - We report measurements of the zero-field resistivity in a dilute 2D electron system in silicon at temperatures down to 35 mK. This extends the previously explored range of temperatures in this system by almost an order of magnitude. On the metallic side, the resistivity near the metal-insulator transition continues to decrease with decreasing temperature and shows no low-temperature upturn. At the critical electron density, the resistivity is found to be temperature independent in the entire temperature range from 35 mK to 1 K. PMID- 11018974 TI - Density of states in coupled chains with off-diagonal disorder AB - We compute the density of states rho(varepsilon) in N coupled chains with random hopping. At zero energy, rho(varepsilon) shows a singularity that strongly depends on the parity of N. For odd N, rho(varepsilon) approximately 1/|varepsilonln (3)varepsilon|, with and without time-reversal symmetry. For even N, rho(varepsilon) approximately |lnvarepsilon| in the presence of time-reversal symmetry, while there is a pseudogap, rho(varepsilon) approximately |varepsilonlnvarepsilon|, in the absence of time-reversal symmetry. PMID- 11018975 TI - Defects, quasibound states, and quantum conductance in metallic carbon nanotubes AB - The effects of impurities and local structural defects on the conductance of metallic carbon nanotubes are calculated using an ab initio pseudopotential method within the Landauer formalism. Substitutionally doped boron or nitrogen produces quasibound impurity states of a definite parity and reduces the conductance by a quantum unit (2e(2)/h) via resonant backscattering. These resonant states show strong similarity to acceptor or donor states in semiconductors. The Stone-Wales defect also produces quasibound states and exhibits quantized conductance reduction. In the case of a vacancy, the conductance shows a much more complex behavior than the prediction from the widely used pi-electron tight-binding model. PMID- 11018976 TI - Dynamic conductance of carbon nanotubes AB - The dynamic conductance of carbon nanotubes was investigated using the nonequilibrium Green's function formalism within the context of a tight-binding model. Specifically, we have studied the ac response of tubes of different helicities, both with and without defects, and an electronic heterojunction. Because of the induced displacement currents, the dynamic conductance of the nanotubes differs significantly from the dc conductance displaying both capacitive and inductive responses. The important role of photon-assisted transport through nanotubes is revealed and its implications for experiments discussed. PMID- 11018977 TI - Coupling between adsorbate vibrations and an electronic surface state AB - We report direct angle-resolved photoemission measurements of the coupling between the symmetric stretch vibrational mode of adsorbed hydrogen and a surface band on W(110). This coupling is manifested by the surface band being split into two branches at a binding energy comparable to the vibrational mode energy, as confirmed by observation of a dramatic hydrogen/deuterium isotope effect. The electron-phonon coupling parameter lambda is found to be significantly larger than that for bulk W, and to be closely related to the degree of surface localization of the surface state wave function. PMID- 11018978 TI - Bloch electrons in a magnetic field: why does chaos send electrons the hard Way? AB - We find that a 2D periodic potential, with different modulation amplitudes in the x and y directions, and a perpendicular magnetic field may lead to a transition to electron transport along the direction of stronger modulation and to localization in the direction of weaker modulation. In the experimentally accessible regime we relate this new quantum transport phenomenon to avoided band crossings due to classical chaos. PMID- 11018979 TI - Apparent spin polarization decay in Cu-dusted Co/Al2O3/Co tunnel junctions AB - Co/Al2O3/Co magnetic tunnel junctions with an interfacial Cu layer have been investigated with in situ growth characterization and ex situ magnetotransport measurements. Cu interlayers grown on Co give an approximately exponential decay of the tunneling magnetoresistance with xi approximately 0.26 nm while those grown on Al2O3 have a decay length of 0.70 nm. The difference in decay lengths can be explained by different growth morphologies, and in this way clarifies a present disagreement in the literature. For monolayer coverage of Cu, we show that the tunneling spin polarization is suppressed by at least a factor of 2 compared to Co and beyond approximately 5 ML it becomes vanishingly small. PMID- 11018980 TI - Conductivity tensor of striped quantum hall phases AB - We study the transport properties of pinned striped quantum Hall phases. We show that, under quite general assumptions, the macroscopic conductivity tensor satisfies a semicircle law. In particular, this result is valid for both smectic and nematic stripe phases, independent of the presence of topological and orientational defects such as dislocations and grain boundaries. As a special case, our results explain the experimental validity of a product rule for the dissipative part of the resistivity tensor, which was previously derived by MacDonald and Fisher (cond-mat/9907278) for a perfect stripe structure. PMID- 11018981 TI - High-field electrical transport in single-wall carbon nanotubes AB - Using low-resistance electrical contacts, we have measured the intrinsic high field transport properties of metallic single-wall carbon nanotubes. Individual nanotubes appear to be able to carry currents with a density exceeding 10(9) A/cm(2). As the bias voltage is increased, the conductance drops dramatically due to scattering of electrons. We show that the current-voltage characteristics can be explained by considering optical or zone-boundary phonon emission as the dominant scattering mechanism at high field. PMID- 11018982 TI - Abrupt change of josephson plasma frequency at the phase boundary of the bragg glass in Bi(2)Sr(2)CaCu(2)O(8+delta) AB - We report the first detailed and quantitative study of the Josephson coupling energy in the vortex liquid, Bragg glass, and vortex glass phases of Bi(2)Sr(2)CaCu(2)O(8+delta) by the Josephson plasma resonance. The measurements revealed distinct features in the T and H dependencies of the plasma frequency omega(pl) for each of these three vortex phases. When going across either the Bragg-to-vortex glass or the Bragg-to-liquid transition line, omega(pl) shows a dramatic change. We provide a quantitative discussion on the properties of these phase transitions, including the first order nature of the Bragg-to-vortex glass transition. PMID- 11018983 TI - Cu NQR study of the stripe phase local structure in the lanthanum cuprates AB - Using Cu nuclear quadrupole resonance (NQR) in Eu-doped La2-xSrxCuO4 we find the evidence of the pinned stripe phase at 1.3 K for 0. 080.18 correlating with the onset of bulk superconductivity corresponds to the depinning of the stripe phase. PMID- 11018984 TI - Magnetothermodynamics of the spin- 1 / 2 Kagome antiferromagnet AB - In this paper, we use a new hybrid method to compute the thermodynamic behavior of the spin- 1 / 2 Kagome antiferromagnet under the influence of a large external magnetic field. We find a T2 low-temperature behavior and a very low sensitivity of the specific heat to a strong external magnetic field. We display clear evidence that this low-temperature magnetothermal effect is associated with the existence of low-lying fluctuating singlets, but also that the whole picture ( T2 behavior of C(v) and the thermally activated spin susceptibility) implies contribution of both nonmagnetic and magnetic excitations. Comparison with experiments is made. PMID- 11018985 TI - Entropy balance and evidence for local spin singlets in a Kagome-like magnet AB - We have measured the specific heat C(T) of the S = 3 / 2 Kagome-lattice containing compound SrCr (9p)Ga (12-9p)O (19). We find little field dependence of the low-temperature C(T), consistent with a low-energy spectrum dominated by many body singlet excitations. At high temperatures, we recover only approximately 50% of the total R ln4 entropy. PMID- 11018986 TI - Thermal and electronic transport properties and two-phase mixtures in La(5/8 x)Pr(x)Ca(3/8)MnO3 AB - We measured thermal conductivity kappa, thermoelectric power S, and electric conductivity sigma of La(5/8-x)Pr(x)Ca(3/8)MnO3, showing an intricate interplay between metallic ferromagnetism (FM) and charge ordering (CO) instability. The change of kappa, S, and sigma with temperature (T) and x agrees well with the effective medium theories for binary metal-insulator mixtures. This agreement clearly demonstrates that with the variation of T as well as x, the relative volumes of FM and CO phases drastically change and percolative metal-insulator transition occurs in the mixture of FM and CO domains. PMID- 11018987 TI - Effects of nuclear spins on the quantum relaxation of the magnetization for the molecular nanomagnet Fe8 AB - The strong influence of nuclear spins on resonant quantum tunneling in the molecular cluster Fe8 is demonstrated for the first time by comparing the relaxation rate of the standard Fe8 sample with two isotopic modified samples: (i) 56Fe is replaced by 57Fe, and (ii) a fraction of 1H is replaced by 2H. By using a recently developed "hole digging" method, we measured an intrinsic broadening which is driven by the hyperfine fields. Our measurements are in good agreement with numerical hyperfine calculations. For T>1.5 K, the influence of nuclear spins on the relaxation rate is less important, suggesting that spin phonon coupling dominates the relaxation rate. PMID- 11018988 TI - Low to high spin-state transition induced by charge ordering in antiferromagnetic YBaCo2O5 AB - The oxygen-deficient double perovskite YBaCo2O5, containing corner-linked CoO5 square pyramids as principal building units, undergoes a paramagnetic to antiferromagnetic spin ordering at 330 K. This is accompanied by a tetragonal to orthorhombic distortion. Below 220 K orbital ordering and long-range Co(2+)/Co(3+) charge ordering occur as well as a change in the Co2+ spin state from low to high spin. This transition is shown to be very sensitive to the oxygen content of the sample. To our knowledge this is the first observation of a spin-state transition induced by long-range orbital and charge ordering. PMID- 11018989 TI - Spatial mapping of mobility and density of the conduction electrons in (FA)2PF6 AB - Novel implementation of the Fourier imaging technique on conduction electron spins in the one-dimensional organic conductors (FA)2PF6 (FA: fluoranthene) is reported. Two-dimensional spatial imaging of resolution 30 &mgr;m(2) is combined with the pulsed-gradient spin-echo technique, to derive maps revealing the local properties of the electron spin density and mobility. The maps generally show pronounced inhomogeneity of both density and mobility on the scale of approximately 30-300 &mgr;m. Highly mobile regions were identified to exist, and the mobility in these was quantitatively evaluated by a basic theoretical model of restricted diffusion. PMID- 11018990 TI - Measurement of the relaxation rate of the magnetization in Mn12O12-acetate using proton NMR echo AB - We present a novel method to measure the relaxation rate W of the magnetization of Mn 12O (12)-acetate (Mn12) magnetic molecular cluster in its S = 10 ground state at low T. It is based on the observation of an exponential growth in time of the proton NMR signal during the thermal equilibration of the magnetization of the molecules. We can explain the novel effect with a simple model which relates the intensity of the proton echo signal to the microscopic reversal of the magnetization of each individual Mn12 molecule during the equilibration process. The method should find wide application in the study of magnetic molecular clusters in off-equilibrium conditions. PMID- 11018991 TI - Impulsive excitation of phonon-pair combination states by second-order raman scattering AB - The excitation of (100)-oriented KTaO3 with 25-fs laser pulses impulsively drives phonon-pair combination states via second-order Raman scattering. Oscillations in the phonon-amplitude covariance at the sum and difference frequency of the two involved phonons are observed in a spectrally and temporally resolved pump-probe experiment. Transmission changes of the sample are dominated by contributions of wave vector conserving phonon-pair combinations from the entire Brillouin zone that have maxima in their combined density of states. For low temperatures the temperature dependence of the covariance oscillations of different phonon combinations is reproduced by a quantum-mechanical model. PMID- 11018992 TI - Observation of vibrational excitation and deexcitation for NO (v = 2) scattering from au(111): evidence for electron-hole-pair mediated energy transfer AB - Measurements of vibrational excitation and deexcitation of NO (v = 2) scattering from a Au(111) surface show that the probability of both processes increases strongly with the kinetic energy of the incident NO. These results are qualitatively consistent with a vibrational energy transfer mechanism involving electron-hole pairs but not with competing adiabatic models. The form of the energy dependence, and other features of the measurements, are not in accord with existing calculations. PMID- 11018993 TI - Scheme for total quantum teleportation AB - We address the issue of totally teleporting the quantum state of an external particle, as opposed to studies on partial teleportation of external single particle states, total teleportation of coherent states and encoded single particle states, and intramolecular teleportation of nuclear spin states. We find a set of commuting observables whose measurement directly projects onto the Bell basis and discuss a possible experiment, based on two-photon absorption, allowing, for the first time, total teleportation of the state of a single external photon through a direct projective measurement. PMID- 11018994 TI - Optimal quantum cloning via stimulated emission AB - We show that optimal universal quantum cloning can be realized via stimulated emission. Universality of the cloning procedure is achieved by choosing systems that have appropriate symmetries. We first discuss a scheme based on stimulated emission in certain three-level systems, e.g., atoms in a cavity. Then we present a way of realizing optimal universal cloning based on stimulated parametric down conversion. This scheme also implements the optimal universal NOT operation. PMID- 11018995 TI - Patterned colloidal deposition controlled by electrostatic and capillary forces. AB - We use substrates chemically micropatterned with anionic and cationic regions to govern the deposition of charged colloidal particles. The direct observation of the colloidal assembly suggests that this process includes two steps: an initial patterned attachment of colloids to the substrate and an additional ordering of the structure upon drying. The driving forces of the process, i.e. , screened electrostatic and lateral capillary interactions, are discussed. This approach makes it possible to fabricate complex, high-resolution two-dimensional arrays of colloidal particles. PMID- 11018996 TI - Instabilities in a liquid-fluidized bed of gas bubbles AB - Gas bubbles in an aqueous foam can be unjammed, or fluidized, by introducing a forced flow of the continuous liquid phase at a sufficiently high rate. We observe that the resulting bubble dynamics are spatially inhomogeneous, exhibiting a sequence of instabilities vs increasing flow rate. First irregular swirls appear, then a single convective roll, and finally a series of stratified convection rolls each with a different average bubble size. PMID- 11018997 TI - Thermodynamics of heat-shock response. AB - Production of heat-shock proteins is induced when a living cell is exposed to a rise in temperature. The heat-shock response of protein DnaK synthesis in E.coli for temperature shifts T-->T+DeltaT and T-->T-DeltaT is measured as a function of the initial temperature T. We observe a reversed heat shock at low T. The magnitude of the shock increases when one increases the distance to the temperature T0 approximately 23 degrees C, thereby mimicking the nonmonotonous stability of proteins at low temperature. This suggests that stability related to hot as well as cold unfolding of proteins is directly implemented in the biological control of protein folding. PMID- 11018998 TI - Role of secondary motifs in fast folding polymers: a dynamical variational principle. AB - A fascinating and open question challenging biochemistry, physics, and even geometry is the presence of highly regular motifs such as alpha helices in the folded state of biopolymers and proteins. Stimulating explanations ranging from chemical propensity to simple geometrical reasoning have been invoked to rationalize the existence of such secondary structures. We formulate a dynamical variational principle for selection in conformation space based on the requirement that the backbone of the native state of biologically viable polymers be rapidly accessible from the denatured state. The variational principle is shown to result in the emergence of helical order in compact structures. PMID- 11018999 TI - Beyond Hebb: exclusive-OR and biological learning. AB - A learning algorithm for multilayer neural networks based on biologically plausible mechanisms is studied. Motivated by findings in experimental neurobiology, we consider synaptic averaging in the induction of plasticity changes, which happen on a slower time scale than firing dynamics. This mechanism is shown to enable learning of the exclusive-OR (XOR) problem without the aid of error backpropagation, as well as to increase robustness of learning in the presence of noise. PMID- 11019000 TI - Particle diffusion in a quasi-two-dimensional bacterial bath. AB - We study the effect of bacterial motion on micron-scale beads in a freely suspended soap film. Given the sizes of bacteria and beads, the geometry of the experiment is quasi-two-dimensional. Large positional fluctuations are observed for beads as large as 10 microm in diameter, and the measured mean-square displacements indicate superdiffusion in short times and normal diffusion in long times. Though the phenomenon is similar to Brownian motions of small particles, its physical origin is different and can be attributed to the collective dynamics of bacteria. PMID- 11019001 TI - Comment on "Stripes and the t-J Model" PMID- 11019002 TI - Hellberg and manousakis reply: PMID- 11019003 TI - Experimental investigation of resonant activation AB - We experimentally investigate the escape from a metastable state over a fluctuating barrier of a physical system. The system is switching between two states under electronic control of a dichotomous noise. We measure the escape time and its probability density function as a function of the correlation rate of the dichotomous noise in a frequency interval spanning more than four frequency decades. We observe resonant activation, namely a minimum of the average escape time as a function of the correlation rate. We detect two regimes in the study of the shape of the escape time probability distribution: (i) a regime of exponential and (ii) a regime of nonexponential probability distribution. PMID- 11019004 TI - Scaling behavior of cyclical surface growth AB - The scaling behavior of cyclical surface growth (e.g., deposition/desorption), with the number of cycles, n, is investigated. The roughness of surfaces grown by two linear primary processes follows a scaling behavior with asymptotic exponents inherited from the dominant process while the effective amplitudes are determined by both. Relevant nonlinear effects in the primary processes may remain so or be rendered irrelevant. Numerical simulations for several pairs of generic primary processes confirm these conclusions. Experimental results for the surface roughness during cyclical electrodeposition/dissolution of silver show a power law dependence on n, consistent with the scaling description. PMID- 11019005 TI - Generalization of the regge-wheeler equation for self-gravitating matter fields AB - It is shown that the dynamical evolution of perturbations on a static spacetime is governed by a standard pulsation equation for the extrinsic curvature tensor. The centerpiece of the pulsation equation is a wave operator whose spatial part is manifestly self-adjoint. In contrast to metric formulations, the curvature based approach to perturbation theory generalizes in a natural way to self gravitating matter fields, including non-Abelian gauge fields and perfect fluids. As an example, the pulsation equations for self-gravitating, non-Abelian gauge fields are explicitly shown to be symmetric. PMID- 11019006 TI - Nonminimal global monopoles and bound orbits AB - We perform a numerical analysis of the gravitational field of a global monopole coupled nonminimally to gravity and find that, for some given nonminimal couplings (in contrast with the minimal coupling case), there is an attractive region where bound orbits exist. We exhibit the behavior of the frequency shifts that would be associated with "rotation curves" of stars in circular orbits in the spacetimes of such global monopoles. PMID- 11019007 TI - Can flavor-independent supersymmetric soft phases Be the source of all CP violation? AB - Recently it has been demonstrated that large phases in softly broken supersymmetric (SUSY) theories are consistent with electric dipole moment constraints, and are motivated in some (type I) string models. Here we consider whether large flavor-independent soft phases may be the dominant (or only) source of all CP violation. In this framework, epsilon and epsilon(')/epsilon can be accommodated, and the SUSY contribution to the B system mixing can be large and dominant. An unconventional flavor structure of the squark mass matrices (with enhanced super-Cabibbo-Kobayashi-Maskawa mixing) is required for consistency with B and K system observables. PMID- 11019008 TI - QCD rescattering and high energy two-body photodisintegration of the deuteron AB - Photon absorption by a quark in one nucleon followed by its high-momentum transfer interaction with a quark in the other may produce two final-state nucleons with high relative momentum. We sum the relevant quark rescattering diagrams to show that the scattering amplitude depends on a convolution between the large angle pn scattering amplitude, the hard photon-quark interaction vertex, and the low-momentum deuteron wave function. The computed absolute values of the cross section are in reasonable agreement with the data. PMID- 11019009 TI - Synchronization of mutually coupled self-mixing modulated lasers AB - Synchronization of mutually coupled chaotic lasers has been demonstrated in a microchip LiNdP4O12 laser array with self-mixing feedback modulation. An abrupt transition to synchronized chaos by way of "phase squeezing" was observed when coupling between the two lasers was increased. This phenomenon is well reproduced by numerical calculations using model equations. It is also shown that low energy variation as well as high disorder are concurrently established in synchronized chaos. PMID- 11019010 TI - Impurity-induced stabilization of solitons in arrays of parametrically driven nonlinear oscillators AB - Chains of parametrically driven, damped pendula are known to support solitonlike clusters of in-phase motion which become unstable and seed spatiotemporal chaos for sufficiently large driving amplitudes. We show that the pinning of the soliton on a "long" impurity (a longer pendulum) expands dramatically its stability region, whereas "short" defects simply repel solitons producing an effective partition of the chain. We also show that defects may spontaneously nucleate solitons. PMID- 11019011 TI - Fingering instabilities of confined elastic layers in tension AB - Fingering instabilities similar to those commonly observed in viscous systems have been observed in purely elastic layers that are strained in tension. The instability is driven by the release of lateral constraints within a confined elastic layer and is observed when the lateral confinement significantly exceeds the thickness of the elastic layer. Our results show convincingly that yielding or flow of a material is not required in order for fingering to be initiated in a confined material. PMID- 11019012 TI - Universality in fragmentation AB - Fragmentation of a two-dimensional brittle solid by impact and "explosion," and a fluid by "explosion" are all shown to become critical. The critical points appear at a nonzero impact velocity, and at infinite explosion duration, respectively. Within the critical regimes, the fragment-size distributions satisfy a scaling form qualitatively similar to that of the cluster-size distribution of percolation, but they belong to another universality class. Energy balance arguments give a correlation length exponent that is exactly one-half of its percolation value. A single crack dominates fragmentation in the slow-fracture limit, as expected. PMID- 11019013 TI - Transverse instability of solitary waves in the generalized kadomtsev petviashvili equation AB - The linear stability of planar solitary waves with respect to long-wavelength transverse perturbations is studied in the framework of the generalized Kadomtsev Petviashvili equation. It is newly discovered that for some nonlinearities in this family, the solitary waves could be transversely unstable even in a medium with negative dispersion. In the case of positive dispersion, they are found to be always unstable. PMID- 11019014 TI - Stable static localized structures in one dimension AB - We study the existence, the stability properties, and the bifurcation structure of static localized solutions in one dimension, near the robust existence of stable fronts between homogeneous solutions and periodic patterns. PMID- 11019015 TI - Observations of the collapse of dilute lyotropic lamellar phases under shear flow AB - Experimental evidence of the collapse of dilute lamellar phases due to shear flow is presented. Two systems are used: one composed of brine and an ionic surfactant, and another composed of water, a nonionic surfactant, and cosurfactant. We observe this transition for a range of lamellar spacings and brine salinity. The results are in reasonable agreement with recent theory in which the suppression of fluctuations by shear plays an important role. PMID- 11019016 TI - Singularity theory study of overdetermination in models for L-H transitions AB - Two dynamical models that have been proposed to describe transitions between low- and high-confinement states in confined plasmas are analyzed using singularity theory and stability theory. It is shown that the stationary-state bifurcation sets have qualitative properties identical to standard normal forms for the pitchfork and transcritical bifurcations. The analysis yields the codimension of the highest-order singularities, from which we find that the unperturbed systems are overdetermined bifurcation problems and derive appropriate universal unfoldings. Questions of mutual equivalence and the character of the state transitions are addressed. PMID- 11019017 TI - Nonlinear theory of nonparaxial laser pulse propagation in plasma channels AB - Nonparaxial propagation of ultrashort, high-power laser pulses in plasma channels is examined. In the adiabatic limit, pulse energy conservation, nonlinear group velocity, damped betatron oscillations, self-steepening, self-phase modulation, and shock formation are analyzed. In the nonadiabatic limit, the coupling of forward Raman scattering (FRS) and the self-modulation instability (SMI) is analyzed and growth rates are derived, including regimes of reduced growth. The SMI is found to dominate FRS in most regimes of interest. PMID- 11019018 TI - Resonant self-trapping and absorption of intense bessel beams AB - We report the observation of resonant self-trapping and enhanced laser-plasma heating resulting from propagation of high intensity Bessel beams in neutral gas. The enhancement in absorption and plasma heating is directly correlated to the spatial trapping of laser radiation. PMID- 11019019 TI - Modification of spatial and temporal gains of stimulated brillouin and raman scattering by polarization smoothing AB - Significant reductions of stimulated Brillouin (SBS) and Raman (SRS) scattering are measured by adding polarization smoothing (PS) to a random phase plate smoothed beam. The associated plasma waves, as measured by Thomson scattering, are spatially and temporally modified and reveal that the interplay between SBS and SRS has to be taken into account to understand the effect of the smoothing. The results also support the numerical simulations [S. Huller et al., Phys. Plasmas 5, 2706 (1998); R. Berger et al., Phys. Plasmas 6, 1043 (1999)] predicting a reduction of self-focusing with PS, resulting in a decrease of the maximum laser intensity and consequently of SBS and SRS gains. PMID- 11019020 TI - Measurement of natural plasma flow along the field lines in the scrape-Off layer on the JT-60U divertor tokamak AB - The plasma profile and parallel plasma flow in the scrape-off layer (SOL) were systematically measured using reciprocating Mach probes installed at the outer midplane and near the divertor magnetic null (x point) in the JT-60U tokamak with a single null divertor. For the ion vertical drift due to the toroidal magnetic field gradient (ion nablaB drift) directed towards the divertor, SOL plasma flow along the magnetic field lines away from the divertor ("flow reversal") was discovered at the midplane far from the divertor. A quantitative evaluation of the ion "Pfirsch-Schluter flow," wherein the parallel flow is naturally produced in a toroidal plasma, was consistent with the measurement. PMID- 11019021 TI - Simultaneous attainment of high electron and ion temperatures in discharges with internal transport barriers in ASDEX upgrade AB - Internal transport barriers have been demonstrated to exist also under conditions with T(e) approximately T(i) approximately 10 keV and predominant electron heating of the tokamak core region. Central electron cyclotron heating was added to neutral beam injection-heated ASDEX Upgrade discharges with a preexisting internal transport barrier, established through programmed current ramping leading to shear reversal. Compared to a reference internal transport barrier discharge without electron cyclotron resonance heating, the electron heat conductivity in the barrier region was found not to increase, in spite of a fivefold increase in electron heat flux, and also angular momentum and ion energy transport did not deteriorate. PMID- 11019022 TI - Adsorption of polyelectrolytes at an oppositely charged surface AB - We develop a scaling theory of polyelectrolyte adsorption at an oppositely charged surface. At low surface charge densities, the thickness of the adsorbed layer is determined by the balance between electrostatic attraction to the charged surface and chain entropy. At high surface charge densities, it is determined by the balance between electrostatic attraction and short-range monomer-monomer repulsion. These different stabilizing mechanisms result in the nonmonotonic dependence of the layer thickness on the surface charge density. PMID- 11019023 TI - Refusing to twist: demonstration of a line hexatic phase in DNA liquid crystals AB - We report conclusive high resolution small angle x-ray scattering evidence that long DNA fragments form an untwisted line hexatic phase between the cholesteric and the crystalline phases. The line hexatic phase is a liquid-crystalline phase with long-range hexagonal bond-orientational order, long-range nematic order, but liquidlike, i.e., short-range, positional order. So far, it has not been seen in any other three dimensional system. By line-shape analysis of x-ray scattering data we found that positional order decreases when the line hexatic phase is compressed. We suggest that such anomalous behavior is a result of the chiral nature of DNA molecules. PMID- 11019024 TI - Boundary effects in chiral polymer hexatics AB - Boundary effects in liquid-crystalline phases can be large due to long-ranged orientational correlations. We show that the chiral-hexatic phase can be locked into an apparent three-dimensional N+6 phase via such effects. Simple numerical estimates suggest that the recently discovered "polymer hexatic" may actually be this locked phase. PMID- 11019025 TI - How fast can cracks propagate? AB - We have performed atomic simulations of crack propagation along a weak interface joining two harmonic crystals. The simulations show that a mode II shear dominated crack can accelerate to the Rayleigh wave speed and then nucleate an intersonic daughter that travels at the longitudinal wave speed. This contradicts the general belief that a crack can travel no faster than the Rayleigh speed. PMID- 11019026 TI - High-pressure elasticity of alpha-quartz: instability and ferroelastic transition AB - The single-crystal elastic moduli of alpha-quartz were measured to above 20 GPa in a diamond-anvil cell by Brillouin spectroscopy. The behavior of the elastic moduli indicates that the high-pressure phase transition in quartz is ferroelastic in nature and is driven by softening of C44 through one of the Born stability criteria. The trends in elastic moduli confirm theoretical predictions, but there are important differences, particularly with respect to the magnitudes of the B(i). The quartz I-II transition occurs prior to complete softening of the mode and amorphization. PMID- 11019027 TI - Progress in monte carlo calculations of fermi systems: normal liquid 3He AB - The application of the diffusion Monte Carlo method to a strongly interacting Fermi system as normal liquid 3He is explored. We show that the fixed-node method together with the released-node technique and a systematic method to analytically improve the nodal surface constitute an efficient strategy to improve the calculation up to a desired accuracy. This methodology shows unambiguously that backflow correlations are enough to generate an equation of state of liquid 3He in excellent agreement with experimental data from equilibrium up to freezing. PMID- 11019028 TI - Dynamics of rough interfaces in chemical vapor deposition: experiments and a model for silica films AB - We study the surface dynamics of silica films grown by low pressure chemical vapor deposition. Atomic force microscopy measurements show that the surface reaches a scale invariant stationary state compatible with the Kardar-Parisi Zhang (KPZ) equation in three dimensions. At intermediate times the surface undergoes an unstable transient due to shadowing effects. By varying growth conditions and using spectroscopic techniques, we determine the physical origin of KPZ scaling to be a low value of the surface sticking probability, related to the surface concentration of reactive groups. We propose a stochastic equation that describes the qualitative behavior of our experimental system. PMID- 11019029 TI - Dense branching morphology in electrodeposition experiments: characterization and mean-field modeling AB - Dense branching morphologies (DBM) obtained in thin gap electrodeposition cells are characterized by a dense array of branches behind a flat advancing envelope. In this Letter, we show the existence in DBM of a new (porous) phase, qualitatively different from a (compact) metal deposit. The local porosity inside the branches is found to be much more robust than geometric characteristics such as the width or the distance between branches. This fact seems to be unreported in previous modeling of DBM. A mean-field model is proposed that displays overall features observed in the experiments, such as concentration profiles, front velocity, and branched internal structure. PMID- 11019030 TI - Unexpected behavior of the local compressibility near the B = 0 metal-insulator transition AB - We have measured the local electronic compressibility of a two-dimensional hole gas as it crosses the B = 0 metal-insulator transition. In the metallic phase, the compressibility follows the mean-field Hartree-Fock (HF) theory and is found to be spatially homogeneous. In the insulating phase it deviates by more than an order of magnitude from the HF predictions and is spatially inhomogeneous. The crossover density between the two types of behavior agrees quantitatively with the transport critical density, suggesting that the system undergoes a thermodynamic change at the transition. PMID- 11019031 TI - Electron-irradiation-induced radiolytic oxygen generation and microsegregation in silicon dioxide polymorphs AB - The first direct in situ observations of the production and microsegregation of radiolytic interstitial oxygen resulting from electron beam irradiation of crystal and amorphous oxygen deficient SiO2 polymorphs has been made using cathodoluminescence (CL) microanalysis (spectroscopy and microscopy). Previously unreported near-infrared CL emission is observed at 0.968+/-0.003 eV from crystal alpha-SiO2 (quartz) and at 0.971+/-0.003 eV from amorphous a-SiO2 (fused quartz and silica glasses) at 290 K. The energy and width of the near-infrared CL emission from electron-irradiated alpha-SiO2 polymorphs is consistent with the O2 (1)Delta(g)-->(3)Sigma(-)(g) transition associated with molecular oxygen. PMID- 11019032 TI - Quantum hall effect at low magnetic fields AB - The temperature and scale dependence of resistivities in the standard scaling theory of the integer quantum Hall effect is discussed. It is shown that recent experiments, claiming to observe a discrepancy with the global phase diagram of the quantum Hall effect, are in fact in agreement with the standard theory. The apparent low-field transition observed in the experiments is identified as a crossover due to weak localization and a strong reduction of the conductivity when Landau quantization becomes dominant. PMID- 11019033 TI - Strong, ultranarrow peaks of longitudinal and hall resistances in the regime of breakdown of the quantum hall effect AB - With unusually slow and high-resolution sweeps of magnetic field, strong ultranarrow (width down to 100 &mgr;T) resistance peaks are observed when high currents are applied through quantum Hall samples. The peaks are dependent on the directions and even the history of magnetic field sweeps, indicating the involvement of a very slow physical process. Such a process and the sharp peaks are, however, not predicted by existing theories. We also find that the sharp resistance peaks are influenced by the nuclear spin flips. PMID- 11019034 TI - Current-driven magnetization reversal and spin-wave excitations in Co /Cu /Co pillars AB - Using thin film pillars approximately 100 nm in diameter, containing two Co layers of different thicknesses separated by a Cu spacer, we examine the process by which the scattering from the ferromagnetic layers of spin-polarized currents flowing perpendicular to the layers causes controlled reversal of the moment direction in the thin Co layer. The well-defined geometry permits a quantitative analysis of this spin-transfer effect, allowing tests of competing theories for the mechanism and also new insight concerning magnetic damping. When large magnetic fields are applied, the spin-polarized current no longer fully reverses the magnetic moment, but instead stimulates spin-wave excitations. PMID- 11019035 TI - Mobile bipolaron AB - We explore the properties of the bipolaron in a 1D Holstein-Hubbard model with dynamical quantum phonons. Using a recently developed variational method combined with analytical strong coupling calculations, we compute correlation functions, effective mass, bipolaron isotope effect, and the phase diagram. The two site bipolaron has a significantly reduced mass and isotope effect compared to the on site bipolaron, and is bound in the strong coupling regime up to twice the Hubbard U naively expected. The model can be described in this regime as an effective t-J-V model with nearest neighbor repulsion. These are the most accurate bipolaron calculations to date. PMID- 11019036 TI - First observation for a cuprate superconductor of fluctuation-induced diamagnetism well inside the finite-magnetic-field regime AB - For the first time for a cuprate superconductor, measurements performed above T(c) in high quality grain aligned La1.9Sr0.1CuO4 samples have allowed the observation of the thermal fluctuation induced diamagnetism well inside the finite-magnetic-field fluctuation regime. These results may be explained in terms of the Gaussian Ginzburg-Landau approach for layered superconductors, but only if the finite field contributions are estimated by taking off the short-wavelength fluctuations. PMID- 11019037 TI - Paramagnetic reentrant effect in high purity mesoscopic AgNb proximity structures AB - We discuss the magnetic response of clean Ag coated Nb proximity cylinders in the temperature range 150 &mgr;K A-AF --> C-AF --> G-AF with increasing of x, is also explained based on the instability of FM states with respect to the spin-wave excitations. PMID- 11019040 TI - Spontaneous plaquette dimerization in the J1-J2 heisenberg model AB - We investigate the nonmagnetic phase of the spin-half frustrated Heisenberg antiferromagnet on the square lattice using exact diagonalization (up to 36 sites) and quantum Monte Carlo techniques (up to 144 sites). The spin gap and the susceptibilities for the most important crystal symmetry breaking operators are computed. A genuine and somehow unexpected "plaquette resonating valence bond," with spontaneously broken translation symmetry and no broken rotation symmetry, comes out from our numerical simulations as the most plausible ground state for J(2)/J(1) approximately 0.5. PMID- 11019041 TI - Abrupt appearance of the domain pattern and fatigue of thin ferroelectric films AB - We study the domain structure in ferroelectric thin films with a "passive" (nonferroelectric) layer at the interface between the film and electrodes. An abrupt transition from a monodomain to a polydomain state has been found with the increase of the passive layer thickness d. The domain width changes very quickly at the transition (exponentially with d(-2)). The slope of the hysteresis loop is in agreement with experiment, assuming realistic parameters of the layer. The slope scales as 1/d, involving only the properties of the layer. We believe that specific properties of the domain structure in ferroelectrics with a passive layer can resolve the long-standing "paradox of the coercive field." PMID- 11019042 TI - Dynamical aspects of the photoinduced phase transition in spin-crossover complexes AB - We report a dynamical study on the photoinduced cooperative changes of the spin configurations in single crystals of the organometal spin-crossover complex. In the photoswitching process between low- and high-spin states, nonlinear characteristics such as thresholdlike behavior, incubation period, and phase separation have been observed. These results demonstrate that the cooperative intersystem crossing mediated by spin-lattice interaction plays a key role in the driving process of a new class of nonequilibrium phenomena so called photoinduced phase transition. PMID- 11019043 TI - Self-organized networks of competing boolean agents AB - A model of Boolean agents competing in a market is presented where each agent bases his action on information obtained from a small group of other agents. The agents play a competitive game that rewards those in the minority. After a long time interval, the poorest player's strategy is changed randomly, and the process is repeated. Eventually the network evolves to a stationary but intermittent state where random mutation of the worst strategy can change the behavior of the entire network, often causing a switch in the dynamics between attractors of vastly different lengths. PMID- 11019044 TI - Recovery of entanglement lost in entanglement manipulation AB - When an entangled state is transformed into another one with probability one by local operations and classical communication, the quantity of entanglement decreases. This Letter shows that entanglement lost in the manipulation can be partially recovered by an auxiliary entangled pair. As an application, a maximally entangled pair can be obtained from two partially entangled pairs with probability one. Finally, this recovery scheme reveals a fundamental property of entanglement relevant to the existence of incomparable states. PMID- 11019045 TI - Interfacial reactions: mixed order kinetics and segregation effects AB - We study A-B reaction kinetics at a fixed interface separating A and B bulks. Initially, the number of reactions R(t) approximately tn(infinity)(A)n(infinity)(B) is second order in the far-field densities n(infinity)(A), n(infinity)(B). First order kinetics, governed by diffusion from the dilute bulk, onset at long times: R(t) approximately x(t)n(infinity)(A), where x(t) approximately t(1/z) is the rms molecular displacement. Below a critical dimension, d0) leads to anomalous decay of interfacial densities. Numerical simulations for z = 2 support the theory. PMID- 11019046 TI - Denoising human speech signals using chaoslike features. AB - A local projective noise reduction scheme, originally developed for low dimensional stationary deterministic chaotic signals, is successfully applied to human speech. This is possible by exploiting properties of the speech signal which resemble structure exhibited by deterministic dynamical systems. In high dimensional embedding spaces, the strong inherent nonstationarity is resolved as a sequence of many different dynamical regimes of moderate complexity. PMID- 11019047 TI - Mean-field solution of the small-world network model. AB - The small-world network model is a simple model of the structure of social networks, which possesses characteristics of both regular lattices and random graphs. The model consists of a one-dimensional lattice with a low density of shortcuts added between randomly selected pairs of points. These shortcuts greatly reduce the typical path length between any two points on the lattice. We present a mean-field solution for the average path length and for the distribution of path lengths in the model. This solution is exact in the limit of large system size and either a large or small number of shortcuts. PMID- 11019049 TI - Comment on "Thermodynamics of black holes: An analogy with glasses" PMID- 11019050 TI - Comment on "Coherent acceleration by subcycle laser pulses" PMID- 11019048 TI - Spontaneous group formation in the seceder model. AB - The seceder model shows how the local tendency to be different gives rise to the formation of groups. The model consists of a population of simple entities which reproduce and die. In a single reproduction event three individuals are chosen randomly and the individual which possesses the largest distance to their center is reproduced by creating a mutated offspring. The offspring replaces a randomly chosen individual of the population. The paper demonstrates the complex group formation behavior and its dependency on the population size. PMID- 11019051 TI - Rau, tajima, and hojo reply: PMID- 11019052 TI - Comment on "First measurement of the rotational constants for the homonuclear molecular ion He( +)(2)" PMID- 11019053 TI - Hardy and wang reply: PMID- 11019054 TI - How to study the elusive efimov state of the 4He3 molecule through a new atom optical state-selection technique AB - Excited states and excitation energies of weakly bound systems, e.g., atomic few body systems and clusters, are difficult to study experimentally. For this purpose we propose a new and very general atom-optical method which is based on inelastic diffraction from transmission gratings. The technique is applicable to the recently found helium trimer molecule 4He3, allowing for the first time an investigation of the possible existence of an excited trimer state and determination of its excitation energy. This would be of fundamental importance for the famous Efimov effect. PMID- 11019055 TI - Calculated equation of state of al, cu, ta, mo, and W to 1000 GPa AB - Calculated Hugoniots and 293-K isotherms at pressures up to 1 TPa (10 Mbar) for the five reference metals Al, Cu, Ta, Mo, and W are reported using the classical mean-field approach where both the cold and the thermal parts of the Helmholtz free energy are derived entirely from the 0-K total energies and electronic density of states calculated with the full-potential linearized augmented plane wave method within the generalized gradient approximation to exchange correlational functional. Our approach permits efficient computation and invokes no empirical parameters. Both the experimental Hugoniots and 293-K isotherms are reproduced excellently. PMID- 11019056 TI - Rapid thermal relaxation in near-critical fluids and critical speeding up: discrepancies caused by boundary effects AB - We present one-dimensional numerical simulations reporting the temperature evolution of a pure fluid subjected to heating near its liquid-vapor critical point under weightlessness. In this model, thermal boundary conditions are imposed at the outer edges of the solids in contact with the fluid. Our investigations concern the thermal conditions at the edges of the fluid and their consequences on the fluid's global response. The results for piston effect heating are shown to be significantly affected by the simulation of the solid boundaries. Concerning critical speeding up, it is even found that taking conductive solids into account can make the bulk fluid temperature change in a way opposed to that predicted in their absence. PMID- 11019057 TI - Convective nonlinearity in non-newtonian fluids AB - In the limit of infinite yield time for stresses, the hydrodynamic equations for viscoelastic, non-Newtonian liquids such as polymer melts must reduce to that for solids. This piece of information suffices to uniquely determine the nonlinear convective derivative, an ongoing point of contention in the rheology literature. PMID- 11019058 TI - Phase coherent vacuum-ultraviolet to radio frequency comparison with a mode locked laser AB - We demonstrate a versatile new technique that provides a phase coherent link between optical frequencies and the radio frequency domain. The regularly spaced comb of modes of a mode-locked femtosecond laser is used as a precise ruler to measure a large frequency gap between two different multiples (harmonics or subharmonics) of a laser frequency. In this way, we have determined a new value of the hydrogen 1S-2S two-photon resonance, f(1S-2S) = 2 466 061 413 187.29(37) kHz, representing now the most accurate measurement of an optical frequency. PMID- 11019059 TI - Discrete breathers and delocalization in nonlinear disordered systems AB - We find exact localized time-periodic solutions with frequencies inside the linearized spectrum [intraband discrete breathers (IDBs)] in random nonlinear models using a new self-consistent method. The IDB frequencies belong to intervals between forbidden gaps generated by resonances with the linear modes, becoming fat Cantor sets in infinite systems. When localized IDBs are continued versus frequency, they delocalize and become multisite IDBs (not predicted by existing theorems), which can propagate energy. Some implications for energy relaxation in glasses are discussed. PMID- 11019060 TI - On the spacing of planetary systems. AB - We present a simplified model of planetary accretion based on conservation of mass, conservation of momentum, and angular-momentum-deficit stability. Within the limitations of this model, we show how the organization of generic planetary systems may be derived from the knowledge of their initial mass distribution. Comparisons with our Solar System and the upsilon-Andromedae planetary system are presented. PMID- 11019061 TI - Stability of halo orbits. AB - We predict new populations of trapped nonequatorial ("halo") orbits of charged dust grains about an arbitrary axisymmetric planet. Simple equilibrium and stability conditions are derived, revealing dramatic differences between positively and negatively charged grains in prograde or retrograde orbits. Implications for the Cassini mission to Saturn are discussed. PMID- 11019062 TI - Lithium-6: A probe of the early universe AB - I consider the synthesis of 6Li due to the decay of relic particles, such as gravitinos or moduli, after the epoch of big bang nucleosynthesis. The synthesized 6Li/H ratio may be compared to 6Li/H in metal-poor stars which, in the absence of stellar depletion of 6Li, yields significantly stronger constraints on relic particle densities than the usual consideration of overproduction of 3He. Production of 6Li during such an era of nonthermal nucleosynthesis may also be regarded as a possible explanation for the relatively high 6Li/H ratios observed in metal-poor halo stars. PMID- 11019063 TI - Chiral-symmetry realization for even- and odd-parity baryon resonances AB - Baryon resonances with even and odd parity are collectively investigated from the viewpoint of chiral symmetry (ChS). We propose a quartet scheme where Delta's and N(*)'s with even and odd parity form a chiral multiplet. This scheme gives parameter-free constraints on the baryon masses in the quartet, which are consistent with observed masses with spin 1 / 2,3 / 2,5 / 2. The scheme also gives selection rules in the one-pion decay: The absence of the parity nonchanging decay N(1720)-->piDelta(1232) is a typical example which should be confirmed experimentally to unravel the role of ChS in baryon resonances. PMID- 11019064 TI - Measurement of differences between J/psi and psi(') suppression in p-A collisions AB - Measurements of the suppression of the yield per nucleon of J/psi and psi(') production for 800 GeV/ c protons incident on heavy nuclear targets, relative to light nuclear targets, have been made with very broad coverage in x(F) and p(T). The observed suppression is smallest at x(F) values of 0.25 and below, and increases at larger values of x(F). It is also strongest at small p(T). Substantial differences between psi(') and J/psi production are observed for the first time in p-A collisions. The suppression for psi(') production is stronger than that for J/psi for x(F) near zero, but becomes comparable to that for J/psi for x(F)>0.6. PMID- 11019065 TI - Effective restoration of the U(A)(1) symmetry in the SU(3) linear sigma model AB - The effective restoration of the chiral U(A)(1) symmetry in strong interactions is studied using the linear chiral SU(3)xSU(3) model at finite temperatures. We find that the disappearance of the chiral anomaly causes a considerable change in the meson mass spectrum. We propose several signals for detecting this chiral phase in ultrarelativistic heavy-ion collisions: The eta/pi(0) ratio is enhanced by an order of magnitude, the a(0) is suppressed in the K&Kmacr; mass spectrum, and the scalar kappa meson appears as a peak just below the K(*)(892) in the invariant piK mass spectrum. PMID- 11019066 TI - Dynamical relativistic effects in quasielastic 1p-shell proton knockout from 16O AB - We have measured the cross section for quasielastic 1p-shell proton knockout in the 16O(e,e(')p) reaction at omega = 0.439 GeV and Q2 = 0.8 (GeV/c)(2) for missing momentum P(miss)infinity; below it, the equilibrium amplitude vanishes. Simulations support these predictions providing new insight into the long-time nonlinear fate of the wave due to Landau damping in plasmas. PMID- 11019080 TI - Steady-state fully noninductive current driven by electron cyclotron waves in a magnetically confined plasma AB - A steady-state, fully noninductive plasma current has been sustained for the first time in a tokamak using electron cyclotron current drive only. In this discharge, 123 kA of current have been sustained for the entire gyrotron pulse duration of 2 s. Careful distribution across the plasma minor radius of the power deposited from three 0. 5-MW gyrotrons was essential for reaching steady-state conditions. With central current drive, up to 153 kA of current have been fully replaced transiently for 100 ms. The noninductive scenario is confirmed by the ability to recharge the Ohmic transformer. The dependence of the current drive efficiency on the minor radius is also demonstrated. PMID- 11019081 TI - Buoyant magnetic flux tubes enhance radiation in Z pinches AB - In numerous experiments, magnetic energy coupled to strongly radiating Z-pinch plasmas exceeds the thermalized kinetic energy, sometimes by a factor of 2-3. We demonstrate that the enhanced energy coupling may be due to the buoyancy rise of toroidal magnetic flux tubes converging to the axis through the unstable pinch plasma. We derive an explicit formula for the enhanced dissipation rate and apply this formula to reconsider an old problem of power balance in a steady-state Z pinch, and then to analyze a new challenge of producing K-shell 3 to 10 keV radiation in long-pulse Z-pinch implosions. PMID- 11019082 TI - Self-acceleration of a tokamak plasma during ohmic H mode AB - Core plasma rotation is observed to change from counter direction to co-current direction during the transition from low (L) to high (H) confinement mode, in Alcator C-Mod plasmas that are heated purely Ohmically and, hence, have no momentum input. The changes of the toroidal velocities, deduced independently from impurity Doppler measurements and from magnetic perturbations associated with sawteeth, agree. The magnitude of the change is consistent with the previously documented scaling for rotation in ion cyclotron rf-heated H modes. The rotation in this Ohmic experiment is obviously not an rf effect but demonstrates unequivocally a transport effect accelerating the plasma. PMID- 11019083 TI - Mixed-mode oscillations and chaos in a glow discharge AB - We present results from numerical simulations on mixed-mode oscillations and chaos excited in a glow discharge, where a model of one-dimensional fluid equations coupled with an external circuit is used. Long duration of high ion and electron densities and fast recharge of a capacitor after a breakdown contribute to the generation of mixed-mode oscillations. The chaotic behavior is characterized by a one-dimensional multibranched map. PMID- 11019084 TI - Molecular dynamics simulation of structural transformation in silicon carbide under pressure AB - Pressure-induced structural transformation in cubic silicon carbide is studied with the isothermal-isobaric molecular-dynamics method using a new interatomic potential scheme. The reversible transformation between the fourfold coordinated zinc-blende structure and the sixfold coordinated rocksalt structure is successfully reproduced by the interatomic potentials. The calculated volume change at the transition and hysteresis are in good agreement with experimental data. The atomistic mechanisms of the structural transformation involve a cubic to-monoclinic unit-cell transformation and a relative shift of Si and C sublattices in the 100 direction. PMID- 11019085 TI - Imaging the elastic properties of coiled carbon nanotubes with atomic force microscopy AB - Coiled carbon nanotubes were produced catalytically by thermal decomposition of hydrocarbon gas. After deposition on a silicon substrate, the three-dimensional structure of the helix-shaped multiwalled nanotubes can be visualized with atomic force microscopy. Helical structures of both chiralities are present in the nanotube deposits. For larger coil diameters ( >170 nm), force modulation microscopy allows one to probe the local elasticity along the length of the coil. Our results agree with the classical theory of elasticity. Similar to the case of straight nanotubes, the Young modulus of coiled multiwalled nanotubes remains comparable to the very high Young modulus of hexagonal graphene sheets. PMID- 11019086 TI - Intrinsic mobility of a dissociated dislocation in silicon AB - Dislocation velocities in silicon in the experimental range of temperature and stress are studied a priori by combining a mechanistic treatment of elementary kink processes with activation energies obtained by atomistic calculations. Pronounced effects of intrinsic coupling of the dissociated partial dislocations are captured in kinetic Monte Carlo simulations, which are consistent with observed velocity variations with applied stress. As a result, the nature of "weak obstacles" to kink propagation, a long-standing postulate in previous data interpretation, is clarified. A striking new effect is predicted and offered for experimental verification when dislocation velocity shows nonmonotonic oscillatory behavior with increasing stress. PMID- 11019087 TI - Hydrodynamic coarsening of binary fluids AB - By suitable interpretation of results from the linear analysis of interface dynamics, it is found that the hydrodynamic growth of the size L of domains that follow spinodal decomposition in fluid mixtures scales with time as L approximately t(alpha), with alpha = 4/7 in the inertial regime. The previously proposed exponent alpha = 2/3 is shown to indicate only the scaling of the oscillatory frequency omega(-2/3) approximately L of the largest structures of the system. The viscous dissipation in the system occurs within a layer of thickness L(d) that also follows a power law of the form L(d) approximately L3/4 in the inertial regime. In the viscous regime the growth is linear in time L approximately t and the dissipative region remains constant L(d) approximately L0. PMID- 11019088 TI - Structure determination of the two-domain ( 1x4) anatase TiO2(001) surface AB - The reconstructed anatase TiO2(001) surface has been investigated by low-energy electron diffraction (LEED), x-ray photoelectron spectroscopy (XPS), and angle resolved mass spectroscopy of recoiled ions (AR-MSRI). Prior investigations have observed or considered only a (1x1) unreconstructed termination for this surface with no detailed structural analysis. Our LEED results indicate a previously unobserved two-domain (1x4) reconstruction after sputtering and annealing the (1x1) surface. The XPS data for this reconstruction indicate the presence of only Ti4+. Simulations of the AR-MSRI experimental data indicate a best fit for a microfaceted surface, revealing both (103) and (1;03) surface planes. PMID- 11019089 TI - Reentrant mound formation in GaAs(001) homoepitaxy observed by ex situ atomic force microscopy AB - A study of the surface morphology of homoepitaxial GaAs(001) by means of ex situ atomic force microscopy in air reveals the reentrance of mounding behavior at low growth temperatures. A transition from statistical roughening to organized mound formation is observed as the growth temperature is reduced. We show by means of growth simulations that the observed morphology is compatible with anisotropic adatom diffusion in the presence of an Ehrlich-Schwoebel barrier. The mechanism leading to this kind of adatom kinetics at low temperatures is interpreted in terms of surfactant acting arsenic condensing on the surface. PMID- 11019090 TI - First principles simulation of a ceramic /Metal interface with misfit AB - The relaxed atomic structure of a model ceramic/metal interface, 222MgO/Cu, is simulated, including lattice constant mismatch, using first principles local density functional theory plane wave pseudopotential methods. The 399-atom computational unit cell contains 36 O and 49 Cu atoms per layer in accordance with the 7/6 ratio of MgO to Cu lattice constants. The atomic layers on both sides of the interface warp to optimize the local bonding. The interface adhesive energy is calculated. The interface electronic structure is found to vary appreciably with the local environment. PMID- 11019091 TI - Oxidation of iron: the relation between oxidation kinetics and oxide electronic structure AB - The test of the validity of the Fromhold-Cook theory of metal oxidation for the O2/Fe system in the tunnel regime is impeded by the growth of a passivating Fe2+/Fe3+ double layer at T150 degrees C, the thermionic emission of electrons is rate limiting for oxygen coverages larger than 13x10(15) atoms/cm(2). PMID- 11019092 TI - Topological approach to Luttinger's theorem and the fermi surface of a kondo lattice AB - A nonperturbative proof of Luttinger's theorem, based on a topological argument, is given for Fermi liquids in arbitrary dimensions. Application to the Kondo lattice shows that even completely localized spins contribute to the Fermi sea volume as electrons, whenever the system can be described as a Fermi liquid. PMID- 11019093 TI - Cyclotron resonance in the layered perovskite superconductor Sr2RuO4 AB - We have observed cyclotron resonance in the layered perovskite superconductor Sr2RuO4. We obtain cyclotron masses for the alpha, beta, and gamma Fermi surfaces of (4.33+/-0.05)m(e), (5.81+/-0. 05)m(e), and (9.71+/-0.2)m(e), respectively. The appreciable differences between these results and those obtained from de Haas-van Alphen measurements are attributable to strong electron-electron interactions in this system. Our findings appear to be consistent with predictions for an interacting Fermi liquid; indeed, semiquantitative agreement is obtained for the electron pockets beta and gamma. PMID- 11019094 TI - 5f-electron localization in PuSe and PuSb AB - Thin layers of PuSb and PuSe were studied by photoelectron spectroscopy. X-ray photoelectron spectroscopy and high-resolution valence-band ultraviolet photoelectron spectroscopy spectra show localization of the 5f states and a low density of states at E(F) in PuSb. In PuSe, which can be classified as a heavy fermion system with low carrier density, we observed three narrow peaks in the valence band, which can be related to the 5f emission. These three features are very sensitive to stoichiometry deviations and disappear for PuSe prepared at T = 77 K. PMID- 11019095 TI - Unified picture of polariton propagation in bulk GaAs semiconductors AB - High-resolution amplitude and phase linear spectroscopy of high-quality bulk GaAs are reported. The detailed structure of the observed full complex transmission is consistently explained by polariton effects on the basis of microscopic calculations. The coupled equations for the excitonic polarization and the light field in the slab configuration are evaluated using appropriate boundary conditions for the electromagnetic field and the excitonic wave function without reference to additional boundary conditions for the macroscopic polarization. PMID- 11019096 TI - Coulomb-induced positive current-current correlations in normal conductors AB - In the white-noise limit current correlations measured at different contacts of a mesoscopic conductor are negative due to the antisymmetry of the wave function (Pauli principle). We show that current fluctuations at capacitive contacts induced via the long range Coulomb interaction due to charge fluctuations in the mesoscopic sample can be positively correlated. The positive correlations are a consequence of the extension of the wave functions into areas near both contacts. As an example we investigate in detail a quantum point contact in a high magnetic field under conditions in which transport is along an edge state. PMID- 11019097 TI - Zero-conductance resonances due to flux states in nanographite ribbon junctions AB - Electronic transport properties through junctions in nanographite ribbons are investigated using the Landauer approach. In the low-energy regime ribbons with zigzag boundary have a single conducting channel of edge states. The conductance as a function of the chemical potential shows a rich structure with sharp dips of zero conductance. Each zero-conductance resonance is connected with a resonant state which can be interpreted as the superposition of two degenerate flux states with Kekule-like current patterns. These zero-conductance dips are connected with a pronounced negative magnetoresistance. PMID- 11019098 TI - Relationship between the noise-induced persistent current and the dephasing rate AB - ac noise in disordered conductors causes both dephasing of the electron wave functions and a dc current around a mesoscopic ring. We demonstrate that the dephasing rate tau(-1)(varphi) in long wires and the dc current , induced by the same noise and averaged over an ensemble of small rings, are connected in a remarkably simple way: tau(varphi) = C(beta)e. Here e is an electron charge, and the constant C(beta) approximately 1 depends on the Dyson symmetry class. The relationship seems to agree reasonably with experiments. This suggests that the two puzzles, anomalously large persistent current and the low-temperature saturation of the dephasing, may have a common solution. PMID- 11019099 TI - Observation of photon-assisted noise in a diffusive normal metal-superconductor junction AB - We report measurements of nonequilibrium noise in a diffusive normal metal superconductor (N-S) junction in the presence of both dc bias and high-frequency ac excitation. We find that the shot noise of a diffusive N-S junction is doubled compared to a normal diffusive conductor. Under ac excitation of frequency nu the shot noise develops features at bias voltages |V| = hnu/(2e), which bear all the hallmarks of a photon-assisted process. Observation of these features provides clear evidence that the effective charge of the current carriers is 2e, due to Andreev reflection. PMID- 11019100 TI - Aging effects in an anderson insulator AB - Aging, commonly observed in glasses, is a manifestation of breakdown of time translational invariance. Here we demonstrate experimentally aging effects in the electronic system of an Anderson insulator. The aging phenomenon in the electron glass appears to be much less sensitive to temperature than in other systems. The differences in the behavior of the electron glass and a spin glass system are discussed in terms of some microscopic differences between the two systems. PMID- 11019101 TI - Optical second harmonic spectroscopy of boron-reconstructed Si(001) AB - Optical second harmonic generation (SHG) spectroscopy is used to probe Si(001) following thermal decomposition of diborane at the surface. Incorporation of boron (B) at second layer substitutional sites at H-free Si(001) intensifies and redshifts the E1 SHG spectral peak, while subsequent H termination further intensifies and blueshifts E1, in sharp contrast to the effect of bulk B doping or nonsubstitutional B. Ab initio pseudopotential and semiempirical tight binding calculations independently reproduce these unique trends, and attribute them to the surface electric field associated with charge transfer to electrically active B acceptors, and rehybridization of atomic bonds. PMID- 11019102 TI - d-band quantum well states AB - Observations of d-band quantum well states are made for atomically uniform Ag films on Fe(100) using angle-resolved photoemission. For increasing film thicknesses, quantum well peaks within the small 4d bandwidth multiply rapidly and merge into a bulklike spectrum at approximately 25 monolayers. An analysis of the peak positions yields a highly accurate bulk band structure of Ag. A very narrow d-band peak width (13 meV) is observed at the band top. PMID- 11019103 TI - Scaling at the chaos threshold for interacting electrons in a quantum Dot AB - The chaotic mixing by random two-body interactions of many-electron Fock states in a confined geometry is investigated. Two regimes are distinguished in the dependence of the typical number of Fock states that are mixed into an eigenstate on the interaction strength V, the excitation energy varepsilon, and the level spacing Delta. In both regimes the number is large (indicating delocalization in Fock space). However, only the large- V regime is described by the golden rule (indicating chaotic mixing). The crossover region is characterized by a maximum in a scaling function that becomes more pronounced with increasing excitation energy. The scaling parameter that governs the transition is (varepsilonV/Delta(2))ln(Delta/V). PMID- 11019104 TI - Infrared hall effect in high- T(c) superconductors: evidence for non-fermi-liquid hall scattering AB - Infrared ( 20-120 and 900-1100 cm(-1)) Faraday rotation and circular dichroism are measured in high- T(c) superconductors using sensitive polarization modulation techniques. Optimally doped YBa2Cu3O7 thin films are studied at temperatures in the range ( 150 the relaxation rate is T independent and approximately p(3), (ii) the slowing down of spin fluctuation is activated with an energy U, which is also a linear function of p(3) and lim U as p-->0 = 8 K; this energy scale could stem only from a single ion anisotropy, and (iii) the p dependence of the dynamical properties is identical below and above p(c), indicating that they are controlled by local excitation. PMID- 11019113 TI - Butterfly hysteresis loop and dissipative spin reversal in the S = 1/2, V15 molecular complex AB - Time resolved magnetization measurements have been performed on a spin 1/2 molecular complex, so-called V15. Despite the absence of a barrier, magnetic hysteresis is observed over a time scale of several seconds. A detailed analysis in terms of a dissipative two-level model is given, in which fluctuations and splittings are of the same energy. Spin-phonon coupling leads to long relaxation times and to a particular "butterfly" hysteresis loop. PMID- 11019114 TI - Mechanisms of decoherence in weakly anisotropic molecular magnets AB - Decoherence mechanisms in crystals of weakly anisotropic magnetic molecules, such as V15, are studied. We show that an important decohering factor is the rapid thermal fluctuation of dipolar interactions between magnetic molecules. A model is proposed to describe the influence of this source of decoherence. Based on the exact solution of this model, we show that at relatively high temperatures, about 0.5 K, the quantum coherence in a V15 molecule is not suppressed and, in principle, can be detected experimentally. Therefore, these molecules may be suitable prototype systems for study of physical processes taking place in quantum computers. PMID- 11019115 TI - Oscillatory decay of magnetization induced by domain-wall stray fields AB - The demagnetization of a hard ferromagnetic layer via the fringing fields of domain walls created by reversing the moment of a neighboring soft ferromagnetic layer is explored experimentally. An unusual oscillatory decay of the magnetic moment of the hard layer is observed using structures in which the demagnetization occurs after a few hundred cycles. This surprising observation is confirmed on a microscopic scale by detailed imaging of the magnetization of the hard layer using high resolution photoemission electron microscopy and by micromagnetic simulations. PMID- 11019116 TI - Coercivity enhancement in exchange biased systems driven by interfacial magnetic frustration AB - We report the temperature and cooling field dependence of the coercivity of exchange biased MnF(2)/Fe bilayers. When the antiferromagnetic surface is in a state of maximum magnetic frustration and the net exchange bias is zero, we observe a strong enhancement of the coercivity, which is proportional to the exchange coupling between the layers. Hence, the coercivity can be tuned in a reproducible and repeatable fashion in the same sample. We propose that a frustrated interface provides local energy minima which effectively pin the propagating domain walls in the ferromagnet, leading to an enhanced coercivity. PMID- 11019117 TI - Surface-enhanced and normal stokes and anti-stokes Raman spectroscopy of single walled carbon nanotubes. AB - Surface enhancement factors of at least 10(12) for the Raman scattering of single walled carbon nanotubes in contact with fractal silver colloidal clusters result in measuring very narrow Raman bands corresponding to the homogeneous linewidth of the tangential C-C stretching mode in semiconducting nanotubes. Normal and surface-enhanced Stokes and anti-Stokes Raman spectra are discussed in the framework of selective resonant Raman contributions of semiconducting or metallic nanotubes to the Stokes or anti-Stokes spectra, respectively, of the population of vibrational levels due to the extremely strong surface-enhanced Raman process, and of phonon-phonon interactions. PMID- 11019118 TI - Coherent control of absorption and polarization decay in a GaAs quantum well: time and spectral domain studies AB - Two phase-locked pulses are used to coherently excite excitonic polarizations. It is shown that the second pulse can either be strongly amplified by taking up energy gained from the destruction of the exciton polarization or can be decreased drastically by giving up all its energy to excitons. Both the temporal and the spectral signatures of the transmitted pulse shapes agree well with model calculations. PMID- 11019119 TI - Theory of resonant rayleigh scattering from semiconductor microcavities: signatures of disorder AB - We develop a self-consistent, microscopic theory of coherent resonant secondary emission from semiconductor microcavities in the normal-mode-coupling regime. Our theory provides a quantitative description of the spectral, temporal, and angular properties of the disorder-induced emission component-resonant Rayleigh scattering-and offers an intuitive physical explanation of emission properties. PMID- 11019120 TI - Multipartite entanglement for continuous variables: A quantum teleportation network AB - We show that one single-mode squeezed state distributed among N parties using linear optics suffices to produce a truly N-partite entangled state for any nonzero squeezing and arbitrarily many parties. From this N-partite entangled state, via quadrature measurements of N-2 modes, bipartite entanglement between any two of the N parties can be "distilled," which enables quantum teleportation with an experimentally determinable fidelity better than could be achieved in any classical scheme. PMID- 11019121 TI - Universal teleportation with a twist AB - We give a transfer theorem for teleportation based on twisting the entanglement measurement. This allows one to say what local unitary operation must be performed to complete the teleportation in any situation, generalizing the scheme to include overcomplete measurements, non-Abelian groups of local unitary operations (e.g., angular momentum teleportation), and the effect of nonmaximally entangled resources. PMID- 11019122 TI - Two state behavior in a solvable model of beta-hairpin folding. AB - Understanding the mechanism of protein secondary structure formation is an essential part of the protein-folding puzzle. Here we describe a simple model for the formation of a beta hairpin, motivated by the fact that folding of a beta hairpin captures much of the basic physics of protein folding. The modeled hairpin is composed of two interacting Gaussian chains with one pairwise (two body) and two many-body interactions. We show that these many-body interactions, arising from side chain packing effects, are responsible for producing an "all-or none" folding transition. We also estimate the (single exponential) folding/unfolding rate via calculating the thermodynamic weight of the "critical" droplet/bubble. PMID- 11019123 TI - Nonequilibrium fluctuations, traveling waves, and instabilities in active membranes. AB - The stability of a flexible fluid membrane containing a distribution of mobile, active proteins (e.g., proton pumps) is shown to depend on the structure and functional asymmetry of the proteins. A stable active membrane is in a nonequilibrium steady state with height fluctuations whose statistical properties are governed by the protein activity. Disturbances are predicted to travel as waves at sufficiently long wavelength, with speed set by the normal velocity of the pumps. The unstable case involves a spontaneous, pump-driven undulation of the membrane, with clumping of the proteins in regions of high activity. PMID- 11019124 TI - Comment on "Physical picture for light emission in scanning tunneling microscopy" PMID- 11019125 TI - Xiao replies: PMID- 11019126 TI - Comment on "Density of states near the mott-hubbard transition in the limit of large dimensions" PMID- 11019127 TI - Kehrein replies: PMID- 11019129 TI - Loutsenko and roubtsov reply: PMID- 11019128 TI - Comment on "Critical velocities in exciton superfluidity" PMID- 11019130 TI - Comment on "Brownian motion of microscopic solids under the action of fluctuating electromagnetic fields" PMID- 11019131 TI - Dorofeyev, fuchs, and gotsmann reply: PMID- 11019132 TI - Linking numbers for self-avoiding loops and percolation: application to the spin quantum hall transition AB - Nonlocal twist operators are introduced for the O(n) and Q-state Potts models in two dimensions which count the numbers of self-avoiding loops (respectively, percolation clusters) surrounding a given point. Their scaling dimensions are computed exactly. This yields many results: for example, the number of percolation clusters which must be crossed to connect a given point to an infinitely distant boundary. Its mean behaves as (1/3sqrt[3] pi) |ln( p(c)-p)| as p-->p(c)-. As an application we compute the exact value sqrt[3]/2 for the conductivity at the spin Hall transition, as well as the shape dependence of the mean conductance in an arbitrary simply connected geometry with two extended edge contacts. PMID- 11019133 TI - Classification of phase transitions in small systems AB - We present a classification scheme for phase transitions in finite systems like atomic and molecular clusters based on the Lee-Yang zeros in the complex temperature plane. In the limit of infinite particle numbers the scheme reduces to the Ehrenfest definition of phase transitions and gives the right critical indices. We apply this classification scheme to Bose-Einstein condensates in a harmonic trap as an example of a higher order phase transition in a finite system and to small Ar clusters. PMID- 11019134 TI - Gravity waves, chaos, and spinning compact binaries AB - Spinning compact binaries are shown to be chaotic in the post-Newtonian expansion of the two-body system. Chaos by definition is the extreme sensitivity to initial conditions and a consequent inability to predict the outcome of the evolution. As a result, the spinning pair will have unpredictable gravitational waveforms during coalescence. This poses a challenge to future gravity wave observatories which rely on a match between the data and a theoretical template. PMID- 11019135 TI - Prospects for gravitational-wave observations of neutron-star tidal disruption in neutron-star-black-hole binaries. AB - For an inspiraling neutron-star-black-hole (NS-BH) binary, we estimate the gravity-wave frequency f(td) at the onset of NS tidal disruption. We model the NS as a tidally distorted, homogeneous, Newtonian ellipsoid on a circular, equatorial geodesic around a Kerr BH. We find that f(td) depends strongly on the NS radius R, and estimate that LIGO-II (ca. 2006-2008) might measure R to 15% precision at 140 Mpc ( approximately 1 event/yr under current estimates). This suggests that LIGO-II might extract valuable information about the NS equation of state from tidal-disruption waves. PMID- 11019136 TI - Dark energy and the cosmic microwave background radiation. AB - We find that current cosmic microwave background anisotropy data strongly constrain the mean spatial curvature of the Universe to be near zero, or, equivalently, the total energy density to be near critical-as predicted by inflation. This result is robust to editing of data sets, and variation of other cosmological parameters (totaling seven, including a cosmological constant). Other lines of argument indicate that the energy density of nonrelativistic matter is much less than critical. Together, these results are evidence, independent of supernovae data, for dark energy in the Universe. PMID- 11019137 TI - Violation of the Greisen-Zatsepin-Kuzmin cutoff: a tempest in a (magnetic) Teapot? Why cosmic ray energies above 10(20) eV may not require new physics. AB - The apparent lack of suitable astrophysical sources for the observed highest energy cosmic rays within approximately 20 Mpc is the "Greisen-Zatsepin-Kuzmin (GZK) paradox." We constrain representative models of the extragalactic magnetic field structure by Faraday rotation measurements; limits are at the microG level rather than the nG level usually assumed. In such fields, even the highest energy cosmic rays experience large deflections. This allows nearby active galactic nuclei (possibly quiet today) or gamma ray bursts to be the source of ultrahigh energy cosmic rays without contradicting the GZK distance limit. PMID- 11019138 TI - Coupling gravitons to matter. AB - Using relationships between open and closed strings, we present a construction of tree-level scattering amplitudes for gravitons minimally coupled to matter in terms of gauge theory partial amplitudes. In particular, we present examples of amplitudes with gravitons coupled to vectors or to a single fermion pair. We also present two examples with massive graviton exchange, as would arise in the presence of large compact dimensions. The gauge charges are represented by flavors of dynamical scalars or fermions. This also leads to an unconventional decomposition of color and kinematics in gauge theories. PMID- 11019139 TI - Linking solar and long baseline terrestrial neutrino experiments. AB - We show that, in the framework of three light neutrino species with hierarchical masses and assuming no fine tuning between the entries of the neutrino mass matrix, one can use the solar neutrino data to obtain information on the element U(e3) of the lepton mixing matrix. Conversely, a measurement of U(e3) in atmospheric or long baseline accelerator or reactor neutrino experiments would help discriminate between possible oscillation solutions of the solar neutrino problem. PMID- 11019140 TI - Coherent lambda-sigma coupling in s-shell hypernuclei AB - It is found that the suppression due to two-body LambdaN-SigmaN coupling solves the overbinding problem in (5)(Lambda)He but it, in turn, causes a severe underbinding in the four-body systems. The shortage of this binding is overcome by introducing explicitly the Lambda-Sigma coupling which is equivalent to the LambdaNN three-body force. This three-body force becomes strong in the 0(+) states of (4)(Lambda)H and (4)(Lambda)He according to the coherently added enhancement. The 0(+)-1(+) splitting in (4)(Lambda)H and (4)(Lambda)He is found partly due to the LambdaN spin-spin interaction and partly due to the Lambda Sigma coupling in the recent Nijmegen soft-core potential. PMID- 11019142 TI - Interpretation of momentum distribution of recoil ions from laser induced nonsequential double ionization AB - We show that the recently reported momentum distribution of recoil ions from laser induced nonsequential double ionization can be interpreted as a combined effect of interelectron correlation and final state field interaction ("Volkov dressing") of the two outgoing electrons. We also find a cutoff formula for the recoil momentum that gives the maximum momenta observed in the experiments. PMID- 11019141 TI - Entry distribution, fission barrier, and formation mechanism of 254102No AB - The entry distribution in angular momentum and excitation energy for the formation of 254No has been measured after the 208Pb(48Ca,2n) reaction at 215 and 219 MeV. This nucleus is populated up to spin 22Planck's over 2pi and excitation energy greater, similar6 MeV above the yrast line, with the half-maximum points of the energy distributions at approximately 5 MeV for spins between 12Planck's over 2pi and 22Planck's over 2pi. This suggests that the fission barrier is greater, similar5 MeV and that the shell-correction energy persists to high spin. PMID- 11019143 TI - Collective multielectron tunneling ionization in strong fields AB - We investigate the quantum mechanical process of two-electron tunneling in strong external electric fields. Numerical solution of a two-electron s-wave model reveals the existence of collective tunneling ionization in a mode where both electrons stay at equal distance from the nucleus. Otherwise the lagging electron is immediately recaptured. The corresponding double ionization rate fails to explain nonsequential multiple ionization in strong-field laser experiments. However, an empirically modified version of the analytical one-electron tunneling rate of Ammosov, Delone, and Krainov agrees with the experiments to a surprising accuracy. The reason for this agreement is presently unknown. PMID- 11019144 TI - Direct observation of the dynamics of electronic excitations in molecules and small clusters AB - Femtosecond time-resolved photoelectron spectroscopy is applied to study relaxation paths of excited states of mass-selected negatively charged clusters. As a first example, the lifetime of an excited state of the carbon trimer anion is measured directly. In addition, the mechanism of the decay, i.e., the configurations of the participating electronic states, is determined from the photoelectron spectra. In general, this method can be used to study all kinds of electronic excitation and relaxation processes in mass-selected nanoparticles. PMID- 11019145 TI - Threshold and linewidth of a mirrorless parametric oscillator AB - We analyze the above-threshold behavior of a mirrorless parametric oscillator based on resonantly enhanced four-wave mixing in a dense atomic vapor. It is shown that, in the ideal limit, an arbitrary small flux of pump photons is sufficient to reach the oscillator threshold. We demonstrate that, due to the large group velocity delays associated with electromagnetically induced transparency, an extremely narrow oscillator linewidth is possible, making a narrow-band source of nonclassical radiation feasible. PMID- 11019146 TI - Asymmetric electron-nuclear dynamics in two-color laser fields: laser phase directional control of photofragments in H+2 AB - Exact non-Born-Oppenheimer numerical solutions of the time-dependent Schrodinger equation for the 1D H+2 molecule in an intense, two-color (omega+2omega) laser field have been obtained. Both electron and proton kinetic energy spectra show spatial, correlated, asymmetric distributions. The calculated spectra exhibit the same unusual correlations as in experiments, in which both positively charged nuclear fragments and negatively charged photoelectrons were preferentially emitted in the same direction. The above asymmetries of photoemission of electrons seen in our quantum simulation are interpreted in the framework of a quasistatic tunneling model. PMID- 11019147 TI - Quantum resonances of the kicked rotor and the SU(q) group AB - The quantum kicked rotor map is embedded into a continuous unitary transformation generated by a time-independent quasi Hamiltonian. In some vicinity of a quantum resonance of order q, we relate the problem to the regular motion along a circle in a (q(2)-1) component inhomogeneous "magnetic" field of a quantum particle with q intrinsic degrees of freedom described by the SU(q) group. This motion is in parallel with the classical phase oscillations near a nonlinear resonance. PMID- 11019148 TI - Discrete breathers: exact solutions in piecewise linear models AB - Exact breather solutions are constructed in piecewise linear (PWL) versions of the discrete nonlinear Schrodinger and Klein-Gordon equations. These solutions correspond to intersections of stable and unstable manifolds of relevant fixed points in associated 2D mappings, an exact construction of which is possible due to the PWL nature of the models. Such exact solutions give us insight into several aspects of breather properties. The problem of dynamical stability of the breathers is mentioned as an instance, detailed results on which will be presented in a future paper. PMID- 11019149 TI - High-efficiency single-mode free-electron maser oscillator based on a bragg resonator with step of phase of corrugation AB - A new type of high-selective Bragg resonator having a step of corrugation inside the interaction region was used as a microwave system for a free-electron maser (FEM). Using a LINAC LIU-3000 (1 MeV/200 A/200 ns) to drive the FEM oscillator, a single-mode single-frequency operation was achieved at a frequency of 30.74 GHz with an output power of about 50 MW, which corresponded to a record efficiency of 35% for a millimeter wavelength FEM. PMID- 11019150 TI - Spatial field distributions in the transition from ballistic to diffusive transport in randomly corrugated waveguides AB - A detailed analysis of the statistical properties of the transmitted field in corrugated waveguides in the transition from ballistic to diffusion transport is presented. The field distributions are found to be highly dependent on the incident mode excited. When the speckle contrast is one, our results agree with recent experiments where the intensity distribution follows the predictions of the random-phasor-sum model, even though the field phase is not uniformly distributed. Clear deviations from the Gaussian statistics are found which can be interpreted in terms of " K" distributions arising either from a small number of scatterers or from perturbations of the speckle pattern on propagation. PMID- 11019151 TI - Catastrophic collapse of ultrashort pulses AB - We investigate theoretically the self-focusing dynamics of an ultrashort laser pulse both near and above the threshold at which the pulse effectively undergoes catastrophic collapse. We find that, as a result of space-time focusing and self steepening, an "optical shock" wave forms inside the medium that gives rise to a broad blueshifted pedestal in the transmitted pulse spectrum. Our results are in good agreement with the primary features observed in experiments and thus provide a theoretical understanding for the underlying process that gives rise to "white light" generation. PMID- 11019152 TI - Raman gap solitons AB - We show that an intense pump pulse, detuned far from the Bragg resonance of a nonlinear periodic structure, can excite a gap soliton at a wavelength within the band gap that corresponds to the Raman shift of the medium. This Raman gap soliton is a stable, long-lived, quasistationary excitation that exists within the grating even after the pump pulse has passed. We find both stationary solitons as well as slow Raman gap solitons with velocities as low as 1% of the speed of light. The predicted phenomena should be observable in fiber Bragg gratings and other nonlinear photonic band gap structures. PMID- 11019153 TI - Benard-Marangoni convection in two-layered liquids AB - We describe experiments on Benard-Marangoni convection in horizontal layers of two immiscible liquids. Unlike previous experiments, which used gases as the upper fluid, we find a square planform close to onset which undergoes a secondary bifurcation to rolls at higher temperature differences. The scale of the convection pattern is that of the thinner lower fluid layer for which buoyancy and surface tension forces are comparable. The wave number of the pattern near onset agrees with the linear stability prediction for the full two-layer problem. The square planform is in qualitative agreement with recent two-layer weakly nonlinear theories, which fail however to predict the transition to rolls. PMID- 11019154 TI - Stationary velocity and charge distributions of grains in dusty plasmas AB - Within the kinetic approach the velocity and the charge distributions of grains in stationary dusty plasmas are calculated and the relations between the effective temperatures of such distributions and plasma parameters are established. It is found that the effective temperature which determines the velocity grain distribution could be anomalously large due to the action of accelerating ionic bombarding force. The possibility to apply the results obtained to the explanation of the increasing grain temperature in the course of Coulomb-crystal melting by reduction of the gas pressure is discussed. PMID- 11019155 TI - Opacity of an underdense plasma slab due to the parametric instabilities of an ultraintense laser pulse AB - The interaction of ultraintense laser beams with underdense plasma slabs has been investigated with two-dimensional particle-in-cell numerical simulations, showing a strong absorption and a correlatively low transmission. Energetic electrons in the multi-MeV range are produced. At very high intensities the plasma transparency is recovered. These results are interpreted in terms of the development of electron parametric instabilities in the self-consistently heated plasma. PMID- 11019156 TI - Impact of kinetic processes on the macroscopic nonlinear evolution of the electromagnetic-beam-plasma instability AB - We present a new, fully kinetic mechanism of generation of spatial magnetic vortices that results from the resonant wave-particle interaction in a plasma. This phenomenon is of basic theoretical interest. It can be responsible for the magnetic vortices observed in numerical simulations in the wake of an ultrastrong, ultraintense laser pulse in an underdense plasma. PMID- 11019157 TI - Scale size of magnetic turbulence in tokamaks probed with 30-MeV electrons AB - Measurements of synchrotron radiation emitted by 30-MeV runaway electrons in the TEXTOR-94 tokamak show that the runaway population decays after switching on neutral beam injection (NBI). The decay starts only with a significant delay, which decreases with increasing NBI heating power. This delay provides direct evidence of the energy dependence of runaway confinement, which is expected if magnetic modes govern the loss of runaways. Application of the theory by Mynick and Strachan [Phys. Fluids 24, 695 (1981)] yields estimates for the "mode width" (delta) of magnetic perturbations: delta<0.5 cm in Ohmic discharges, increasing to delta = 4.4 cm for 0. 6 MW NBI. PMID- 11019158 TI - Jets in tokamaks: A theoretical study AB - A new class of bifurcation of the momentum balance equations for a tokamak plasma is presented. The solution exhibits a monopolar localized jet of ExB flow. The jet is generated by the reduction of turbulent viscosity due to ExB velocity shear. Strong jets of localized plasma flow have been observed in tokamaks as a precursor to the development of a transport barrier region with reduced turbulent transport. The jet solution is shown to fit well with the experimental observations. PMID- 11019159 TI - Generation of broadband electrostatic waves in Earth's magnetotail. AB - The theory that broad-band electrostatic waves (BEN) in Earth's magnetotail are trapped-electron ("BGK") modes is reexamined. Electron/ion beams analyzed for a realistic magnetized-plasma source model with kappa distributions are found to drive an unstable spectrum of broad angular range over several orders of magnitude in f, up to (0.1-0.2)f(pe). Analysis indicates that trapping essential for the BGK paradigm is good only at the highest f, whereas most of the spectrum has minimal trapping and can be driven by electron/ion beam instabilities. A new model is proposed in which trapped-electron modes exist only at the highest f band, whereas electron/ion beam instabilities drive the bulk of the broad-band spectrum below that. BEN wave data from ISEE-1 and ISEE-3 show large angles of propagation with respect to the magnetic field for ff(ce) is observed only in a narrow angular range around the magnetic field and may be BGK modes. This predicts that the BEN solitary waves in the source region are not in BEN well into the lobe. PMID- 11019160 TI - Electron holography with atomic focusers AB - In a modified form of electron holography, as originally proposed by Gabor, a specimen illuminated by the focused, convergent beam of a scanning transmission electron microscope is followed by a thin crystal which acts as a periodic array of atomic focusers. Each of the broad diffraction spots of the crystal then contains a magnified image of the specimen with a resolution limit of 0.05 nm or less. The method is illustrated by images of crystal lattice planes and tungsten atoms in the diffraction patterns formed by crystals in the walls of carbon nanoshells. PMID- 11019161 TI - Structure of liquid Y3Al5O12 (YAG) AB - The total structure factor S(Q) and the radial distribution function G(r) of liquid Y3Al5O12 (YAG) were measured at 1770-2230 K by x-ray scattering from samples under containerless conditions in Ar and O2. Nominal coordination numbers are 4 for Al3+ and 6 for Y3+ ions. The G(r) has peaks at r approximately 1.8 A for Al-O, r approximately 2. 25 A for Y-O, and r approximately 3.3-3.6 A assigned to metal ions in adjacent AlO5-4 and YO9-6 polyhedral ions. Relative to the pure oxides, G(r) for molten YAG has smaller half-widths for the Al-O and Y-O peaks, and an increased sensitivity to temperature and the ambient gas composition. PMID- 11019162 TI - Gas-liquid transition in a two-dimensional system of millimeter-sized like charged metal balls AB - Metal balls with a diameter of 1.59 mm, gently rubbed against a dielectric surface using a shaker, are seen to spontaneously exhibit a two-dimensional liquidlike order with macroscopic dimensions, viz., interball distances of several millimeters. This liquidlike order transforms to a gaslike order through coexistence upon decreasing the area fraction of the balls. The measured pair interaction of like-charged balls surprisingly exhibits a long-range attractive term analogous to that in charged colloids. PMID- 11019163 TI - Experimental evidence for fast heterogeneous collective structural relaxation in a supercooled liquid near the glass transition AB - We have extended the exploration of microscopic dynamics of supercooled liquids to small wave numbers Q corresponding to the scale of intermediate range order, by developing a new experimental approach for precise data correction for multiple scattering noise in inelastic coherent neutron scattering. Our results in supercooled Ca0.4K0.6(NO3)(1.4) reveal the first direct experimental evidence, after a decade of controversy, that the so-called picosecond process around the glass transition corresponds to a predicted first, faster stage of the structural relaxation. In addition, they show that this process takes the spatial form of fast heterogeneous collective flow of correlated groups of atoms. PMID- 11019164 TI - Investigation of the radial compression of carbon nanotubes with a scanning probe microscope AB - Efforts have been made to characterize the mechanical properties of carbon nanotubes. Previous work has concentrated on the tubes' longitudinal properties, and studies of their radial properties have lagged behind. We have used a scanning probe microscope with an indentation/scratch function to investigate the radial compression of multiwalled carbon nanotubes under an asymmetric stress. In particular, we have determined the radial compressive elastic modulus at different compression levels and have estimated the compressive strength to be well beyond 5.3 GPa. PMID- 11019165 TI - Quasi-Ab initio molecular dynamic study of Fe melting AB - We have investigated the melting of hcp Fe at high pressure by employing molecular dynamics simulations in conjunction with the full potential linear muffin tin orbital method. Apart from being of fundamental value, the melting of iron at high pressure is also important for our understanding of the Earth. The subject of iron melting at high pressures is controversial. The experimental data for the iron melting temperature can be separated into two regions, "low" and "high." Here we present an ab initio simulated iron melting curve which is in agreement with the low temperatures at lower pressures, but is in excellent agreement with the high-mostly shockwave-temperatures at high pressures. A comparison with available data lends support to the presented iron melting curve. PMID- 11019166 TI - Two experimental tests of the halperin-lubensky-Ma effect at the nematic-smectic- A phase transition AB - We have conducted two quantitative tests of predictions based on the Halperin Lubensky-Ma (HLM) theory of fluctuation-induced first-order phase transitions. First, we explore the effect of an external magnetic field on the nematic-smectic A transition in a liquid crystal. Second, we examine the dependence of the first order discontinuity as a function of mixture concentration in pure 8CB and three 8CB-10CB mixtures. We find the first quantitative evidence for deviations from the HLM theory. PMID- 11019167 TI - Imaging cluster surfaces with atomic resolution: the strong metal-support interaction state of Pt supported on TiO2(110) AB - Nanosized platinum clusters were grown on a TiO2(110) surface and annealed in ultrahigh vacuum at high temperatures. This leads to the so-called strong metal support interaction (SMSI) state, characterized by a complete encapsulation of the clusters with a reduced titanium oxide layer. We present atomically resolved scanning tunneling microscopy measurements of the cluster surfaces and an atomic model of the SMSI state. The ability to resolve the cluster surface geometry with atomistic detail may help to identify the active sites responsible for the SMSI. PMID- 11019168 TI - Effect of the environment on alpha-Al2O3 (0001) surface structures AB - We report that calculating the Gibbs free energy of the alpha-Al2O3 (0001) surfaces in equilibrium with a realistic environment containing both oxygen and hydrogen species is essential for obtaining theoretical predictions consistent with experimental observations. Using density-functional theory we find that even under conditions of high oxygen partial pressure the metal-terminated surface is surprisingly stable. An oxygen-terminated alpha-Al2O3 (0001) surface becomes stable only if hydrogen is present on the surface. In addition, including hydrogen on the surface resolves discrepancies between previous theoretical work and experimental results with respect to the magnitude and direction of surface relaxations. PMID- 11019169 TI - Indium-indium pair correlation and surface segregation in InGaAs alloys AB - In-In pair correlations and In surface segregation in In xGa 1-xAs alloys are studied by first-principles total-energy calculations. By calculating the substitution energy of a single In atom, we find that the near-surface energetics explains the observed In segregation on InGaAs(001)-beta2(2x4) surfaces. Indium surface segregation further enhances the In site selectivity, thus the long-range ordering. We find that the [110] and [001] In-In pair correlations are repulsive and nearly isotropic in bulk but are highly anisotropic near the (001) surface. The sign of the [110] In-In interaction energies vs the distance from the surface is oscillatory. These findings explain the recent puzzling cross-sectional x-STM results. PMID- 11019170 TI - Direct local epitaxy of diamond on Si(100) and surface-roughening-induced crystal misorientation AB - A direct diamond epitaxy on the silicon substrate is demonstrated not only at the interface formed during the growth process but also at the nucleation sites. The small (001) terraces with dimensions of several atomic distances at the site of nucleation are formed due to the roughening of silicon surface and lead to the grain misorientation. A model is presented which attempts to explain the initial stages of diamond growth. Predictions are made for methods of improving the nucleation of epitaxial diamond crystallites. PMID- 11019171 TI - Decay of silicon mounds: scaling laws and description with continuum step parameters AB - The decay of mounds about a dozen layers high on the Si(111)-(7x7) surface has been measured quantitatively by scanning tunneling microscopy and compared with analytic predictions for the power-law dependence on time predicted for a step mediated decay mechanism. Conformably, we find an exponent 1/4 associated with the (3D) decay of the mound height and exponent 1/3 associated with the (2D) decay of top-layer islands. Using parameters from a continuum step model, we capture the essence of the kinetics. Qualitative features distinguish these mounds from multilayer islands found on metals. PMID- 11019172 TI - Evidence for phase-separated quantum dots in cubic InGaN layers from resonant raman scattering AB - The emission of light in the blue-green region from cubic InxGa1-xN alloys grown by molecular beam epitaxy is observed at room temperature and 30 K. By using selective resonant Raman spectroscopy (RRS) we demonstrate that the emission is due to quantum confinement effects taking place in phase-separated In-rich quantum dots formed in the layers. RRS data show that the In content of the dots fluctuates across the volume of the layers. We find that dot size and alloy fluctuation determine the emission wavelengths. PMID- 11019173 TI - Ground state theory of delta-Pu AB - Correlation effects are important for making predictions in the delta phase of Pu. Using a realistic treatment of the intra-atomic Coulomb correlations we address the long-standing problem of computing ground state properties. The equilibrium volume is obtained in good agreement with experiment when taking into account Hubbard U of the order 4 eV. For this U, the calculation predicts a 5f(5) atomiclike configuration with L = 5, S = 5/2, and J = 5/2 and shows a nearly complete compensation between spin and orbital magnetic moments. PMID- 11019174 TI - Pseudogap formation in the intermetallic compounds (Fe1-xVx)3Al AB - Optical conductivity data of the intermetallic compounds (Fe1-xVx)3Al ( 01 in the limit D2-->0. PMID- 11019179 TI - Calculation of the inelastic scanning tunneling image of acetylene on Cu(100) AB - A Green function linear combination of atomic orbitals technique is used to theoretically calculate the "inelastic" scanning tunneling microscope image of a C2H2 molecule adsorbed on Cu(100) and explain previous experimental results. Our analysis of the inelastic scattering process in terms of the orbitals shows that a destructive interference occurs in the inelastic scattering by the C-H bending modes. This results in a much smaller inelastic fraction due to the bending modes as compared to the stretching ones, and explains why the former cannot be observed experimentally. PMID- 11019180 TI - Transient vortex states in Bi2Sr2CaCu2O(8+delta) crystals AB - A high temporal resolution magneto-optical system is employed to observe the time evolution of the vortex structure in Bi(2)Sr(2)CaCu(2)O(8+delta) crystals after a sudden application of a magnetic field. The magneto-optical images reveal dynamic coexistence of two vortex phases: a quasiordered phase in the sample interior and a transient disordered phase near the sample edges. The border between these two phases, marked by an abrupt change in the gradient of the local induction, moves with time. This motion enables tracing the decay of the transient state and the concurrent growth of the thermodynamic vortex phases. The growth rate is sensitive to the location in the field-temperature phase diagram. PMID- 11019181 TI - Enhanced vortex damping by eddy currents in superconductor-semiconductor hybrids AB - An enhancement of vortex-motion damping in thin Pb/In superconducting films is obtained through coupling to an adjacent two-dimensional electron gas formed in a modulation-doped GaAs/AlGaAs heterostructure. This effect is observed by monitoring the power dissipation in the superconductor in the vortex state while increasing the density of the electron gas using a gate voltage. Quantitative agreement is found with calculations based on a viscous damping model which considers generation of eddy currents in the electron gas by moving flux lines. In the regime of filamentary vortex flow, eddy-current damping leads to a striking dissipation breakdown due to the stopping of entire vortex channels. PMID- 11019182 TI - Critical points in heavy ion irradiated untwinned YBa(2)Cu3O(7-delta) crystals AB - The critical points in untwinned YBa(2)Cu(3)O(7-delta) crystals with dilute columnar defects are investigated. We find a convergence of a first order vortex melting line with an irreversibility line associated with the onset of the Bose glass critical regime at the lower critical point. In addition, we find that columnar defects raise the upper critical point, implying that vortex line meandering is a basic feature controlling its position. PMID- 11019183 TI - Transmission phase shift of a quantum dot with kondo correlations AB - We study the effects of Kondo correlations on the transmission phase shift of a quantum dot in an Aharonov-Bohm ring. We predict in detail how the development of a Kondo resonance should affect the dependence of the phase shift on transport voltage, gate voltage, and temperature. This system should allow the first direct observation of the well-known scattering phase shift of pi/2 expected (but not directly measurable in bulk systems) at zero temperature for an electron scattering off a spin- 1 / 2 impurity that is screened into a singlet. PMID- 11019184 TI - Ferromagnetic, A-type, and charge-ordered CE-type states in doped manganites using jahn-teller phonons AB - The two-orbital model for manganites with both noncooperative and cooperative Jahn-Teller phonons is studied at hole density x = 0.5 using Monte Carlo techniques. The phase diagram is obtained by varying the electron-phonon coupling and the t(2g)-spins exchange. The insulating CE-type charge- and orbital-ordered state with the z-axis charge stacking observed in narrow-bandwidth manganites is stabilized in the simulations. Its charge gap Delta(CO) is much larger than the critical temperature k(B)T(CO). Metalliclike A-type and ferromagnetic states are also obtained in the same framework, and the phase boundaries among them have first-order characteristics. PMID- 11019185 TI - Local spin resonance and spin-peierls-like phase transition in a geometrically frustrated antiferromagnet AB - Inelastic magnetic neutron scattering reveals a localized spin resonance at 4.5 meV in the ordered phase of the geometrically frustrated cubic antiferromagnet ZnCr2O4. The resonance develops abruptly from quantum critical fluctuations upon cooling through a first order transition to a co-planar antiferromagnet at T(c) = 12. 5(5) K. We argue that this transition is a three dimensional analog of the spin-Peierls transition. PMID- 11019186 TI - Polarization instability in polydomain ferroelectric epitaxial thin films and the formation of heterophase structures AB - A thermodynamic theory is developed for dense laminar domain structures in epitaxial ferroelectric films. It is found that, at some critical misfit strain between the film and substrate, the 90 degrees c/a/c/a domain structure becomes unstable with respect to the appearance of the polarization component parallel to domain walls, which results in the formation of a heterophase structure. For PbTiO3 and BaTiO3 films, the stability ranges of polydomain and heterophase states are determined. Dielectric anomalies accompanying misfit-strain-driven structural transformations are described. PMID- 11019187 TI - Novel features of Sr1-xCaxTiO3 phase diagram: evidence for competing antiferroelectric and ferroelectric interactions AB - Structural and dielectric evidences are advanced to show that the Sr1-xCaxTiO3 (SCT) system undergoes an antiferroelectric (AFE) phase transition in the composition range 0.18or = square root[B(c)/3B]. The outflow proposed as input to GRB fireball models. PMID- 11019198 TI - Constraining antimatter domains in the early universe with big bang nucleosynthesis. AB - We consider the effect of a small-scale matter-antimatter domain structure on big bang nucleosynthesis and place upper limits on the amount of antimatter in the early universe. For small domains, which annihilate before nucleosynthesis, this limit comes from underproduction of 4He. For larger domains, the limit comes from 3He overproduction. Since most of the 3He from &pmacr; 4He annihilation are themselves annihilated, the main source of primordial 3He is the photodisintegration of 4He by the electromagnetic cascades initiated by the annihilation. PMID- 11019199 TI - Observational evidence for self-interacting cold dark matter AB - Cosmological models with cold dark matter composed of weakly interacting particles predict overly dense cores in the centers of galaxies and clusters and an overly large number of halos within the Local Group compared to actual observations. We propose that the conflict can be resolved if the cold dark matter particles are self-interacting with a large scattering cross section but negligible annihilation or dissipation. In this scenario, astronomical observations may enable us to study dark matter properties that are inaccessible in the laboratory. PMID- 11019200 TI - Search for neutrino oscillations at the palo verde nuclear reactors AB - We report on the initial results from a measurement of the antineutrino flux and spectrum at a distance of about 800 m from the three reactors of the Palo Verde Nuclear Generating Station using a segmented gadolinium-loaded scintillation detector. We find that the antineutrino flux agrees with that predicted in the absence of oscillations excluding at 90% C.L. nu;(e)-nu;(x) oscillations with Deltam(2)>1.12x10(-3) eV(2) for maximal mixing and sin (2)2straight theta>0.21 for large Deltam(2). Our results support the conclusion that the atmospheric neutrino oscillations observed by Super-Kamiokande do not involve nu(e). PMID- 11019201 TI - Further search for the decay K+-->pi(+)nunu; AB - A search for additional evidence for the rare kaon decay K+-->pi(+)nunu; has been made with a new data set comparable in sensitivity to the previous exposure that produced a single event. No new events were found in the pion momentum region examined, 211pi(+)nunu;) = 1.5(+3.4)( 1.2)x10(-10). PMID- 11019202 TI - Hydrodynamics of nuclear matter in the chiral limit AB - Using the Poisson bracket method, we construct the hydrodynamics of nuclear matter in the chiral limit, which describes the dynamics of all low-energy degrees of freedom, including the fluid-dynamical and pionic ones. The hydrodynamic equations contain, beside five Euler equations of relativistic fluid dynamics, N(2)(f)-1 second order equations describing propagating pions and N(2)(f)-1 first order equations describing the advection of the vector isospin charges. We present hydrodynamic arguments showing that the pion velocity vanishes at the second order phase transition at N(f) = 2. PMID- 11019203 TI - Proton-neutron mixed-symmetry 3(+)(ms) state in 94Mo AB - We identify a Jpi = 3(+)(ms) state in 94Mo. This identification is based on six M1 and E2 strengths and is the first identification of a 3(+)(ms) state from B(M1) and B(E2) values. The transition strengths were determined from the measurement of Doppler shifts, branching ratios, and E2/M1 mixing ratios, obtained from gammagamma directional correlations following the 91Zr(alpha,n) reaction and the beta(+) decay of (94)Tc(m). The interacting boson model agrees with the observations, which prove the 2(+) mixed-symmetry states to be a building block in nuclear structure. PMID- 11019205 TI - Matrix elements of random operators and discrete symmetry breaking in nuclei AB - It is shown that several effects are responsible for deviations of the intensity distributions from the Porter-Thomas law. Among these are genuine symmetry breaking, such as isospin, the nature of the transition operator, truncation of the Hilbert space in shell model calculations, and missing transitions. PMID- 11019204 TI - Search for isovector giant monopole resonances via the Pb(3He,tp) reaction AB - The (nat)Pb(3He,tp) reaction at E(3He) = 177 MeV was studied to identify 2Planck's over 2piomega isovector monopole strength in Bi isotopes. Monopole strength was found in the region -45,omega) of diamond and LiF have been measured using inelastic x-ray scattering. The experimental data are compared to results of ab initio calculations, which take into account the interaction of the excited electron with the remaining hole. In diamond, the vicinity of the indirect band gap and its momentum dependence are studied. In LiF, a larger energy range, which covers the fundamental exciton, the plasmon, and several interband transitions, is investigated. Calculations and measurements agree quite well and emphasize the need to properly include the interaction of the excited electron in the conduction band with the hole in the valence band. PMID- 11019237 TI - Critical fluctuations and quenched disordered two-dimensional charge stripes in La(5/3)Sr(1/3)NiO4 AB - Using high-resolution x-ray scattering, we have demonstrated the existence of quenched disordered charge stripes in a single crystal of La (5/3)Sr (1/3)NiO (4) at low temperatures. Above the second-order transition critical scattering was observed due to fluctuations into the charge stripe phase. The charge stripes are shown to be two dimensional in nature both by measurements of their correlation lengths (xi(a) approximately 185 A, xi(b) = 400 A, and xi(c) approximately 25 A) and by the critical exponents of the charge strip transition. The charge stripe ordering did not develop long-range order even at low temperatures, indicating that the charge stripes are disordered and that the length scale of the disorder is quenched. PMID- 11019238 TI - Topology dependent quantities at the anderson transition AB - The boundary condition dependence of the critical behavior for the three dimensional Anderson transition is investigated. A strong dependence of the scaling function and the critical conductance distribution on the boundary conditions is found, while the critical disorder and critical exponent are found to be independent of the boundary conditions. PMID- 11019239 TI - Stripe conductivity in La1.775Sr0.225NiO4 AB - We report Raman light-scattering and optical conductivity measurements on a single crystal of La1.775Sr0.225NiO4 which exhibits incommensurate charge-stripe order. The extra phonon peaks induced by stripe order can be understood in terms of the energies of phonons that occur at the charge-order wave vector Q(c). A strong Fano antiresonance for a Ni-O bond-stretching mode provides clear evidence for finite dynamical conductivity within the charge stripes. PMID- 11019240 TI - Effects of strain and local charge on the formation of deep defects in III-V ternary alloys AB - The effects of external and internal strains and of defect charges on the formation of gallium vacancies and arsenic antisites in GaAs and In0.5Ga0.5As have been investigated by ab initio density functional methods. Present results show that a proper understanding of strain and defect charge permits the development of a defect engineering of semiconductors. Specifically, they predict that arsenic antisites in InGaAs ternary alloys can form, upon p-type doping in the presence of an arsenic overpressure, even in the case of high-temperature epitaxial growths. PMID- 11019241 TI - Search for two-scale localization in disordered wires in a magnetic field AB - A recent paper [A. V. Kolesnikov and K. B. Efetov, Phys. Rev. Lett. 83, 3689 (1999)] predicts a two-scale behavior of wave function decay in disordered wires in the crossover regime from preserved to broken time-reversal symmetry. We have tested this prediction by a transmission approach, relying on the Borland conjecture that relates the decay length of the transmittance to the decay length of the wave functions. Our numerical simulations show no indication of two-scale behavior. PMID- 11019242 TI - Density of states of disordered two-dimensional crystals with half-filled band AB - A diagrammatic method is applied to study the effects of commensurability in two dimensional disordered crystalline metals by using the particle-hole symmetry with respect to the nesting vector P0 = +/-pi / a,pi / a for a half-filled electronic band. The density of electronic states (DoS) is shown to have nontrivial quantum corrections due to both nesting and elastic impurity scattering processes, and as a result the Van Hove singularity is preserved in the center of the band. However, the energy dependence of the DoS is significantly changed. A small offset from the middle of the band gives rise to the disappearance of quantum corrections to the DoS. PMID- 11019243 TI - Disorder and interaction-induced pairing in the addition spectra of quantum dots AB - We have numerically investigated the electron addition spectra in quantum dots containing a small number (N approximately L-5/3 and L-2.2 for (1+1) and (2+1) dimensional manifolds with random bond disorder. PMID- 11019256 TI - Asymmetric magnetization reversal in exchange-biased hysteresis loops AB - Polarized neutron reflectometry is used to probe the in-plane projection of the net-magnetization vector M--> of polycrystalline Fe films exchange coupled to twinned (110) MnF (2) or FeF (2) antiferromagnetic (AF) layers. The magnetization reversal mechanism depends upon the orientation of the cooling field with respect to the twinned microstructure of the AF, and whether the applied field is increased to (or decreased from) a positive saturating field; i.e. , the magnetization reversal is asymmetric. The reversal of the sample magnetization from one saturated state to the other occurs via either domain wall motion or magnetization rotation on opposite sides of the same hysteresis loop. PMID- 11019257 TI - Influence of low-symmetry distortions on the luminescence of Cr4+-doped forsterite AB - By using pressure to vary the extent of nontetrahedral distortions of the Cr4+ site in Mg2SiO4, we reveal the important influence of the 3T1 state on the emission properties of the 3T2 state. We find that 3T1-3T2 mixing has a pronounced effect on the line shape and radiative decay rate of emission from the 3T2 state and that the extent of mixing depends critically on the magnitude of nontetrahedral distortions. The results provide an explanation for the wide variation of Cr4+ emission properties in different host lattices at ambient pressure and indicate that the tailoring of asymmetric distortions of luminescent centers represents an effective new strategy for tuning the linewidth of spectral transitions. PMID- 11019258 TI - Photon emission spectroscopy of individual oxide-supported silver clusters in a scanning tunneling microscope AB - Photon emission spectra of individual alumina-supported silver clusters have been measured for the first time. The light emission stimulated by electron injection from the tip of a scanning tunneling microscope can be assigned to the (1,0) mode of the Mie-plasmon resonance in small silver particles. As cluster sizes decrease, the resonance position shifts to higher energies and the linewidth increases. In the size range examined (1.5-12 nm), intrinsic size effects are discussed as possible origins for the observed size dependence of the Mie resonance. PMID- 11019259 TI - Angular distribution of electron temperature and density in a laser-ablation plume AB - The angular distribution of electron temperature and density in a laser-ablation plume has been studied for the first time. The electron temperature ranges from 0.1 to 0.5 eV and is only weakly dependent on the angle in the low-intensity range studied here. In contrast, the typical ion energy is about 2 orders of magnitude larger, and its angular distribution is more peaked about the target normal. The derived values of the electron density are in agreement with the measured values of ion density. PMID- 11019260 TI - Entanglement purification of gaussian continuous variable quantum states AB - We describe an entanglement purification protocol to generate maximally entangled states with high efficiencies from two-mode squeezed states or from mixed Gaussian continuous entangled states. The protocol relies on a local quantum nondemolition measurement of the total excitation number of several continuous variable entangled pairs. We propose an optical scheme to do this kind of measurement using cavity enhanced cross-Kerr interactions. PMID- 11019261 TI - Self-organized criticality in the olami-feder-christensen model AB - A system is in a self-organized critical state if the distribution of some measured events obeys a power law. The finite-size scaling of this distribution with the lattice size is usually enough to assume that the system displays self organized criticality. This approach, however, can be misleading. In this paper we analyze the behavior of the branching rate sigma of the events to establish whether a system is in a critical state. We apply this method to the Olami-Feder Christensen model to obtain evidence that, in contrast to previous results, the model is critical in the conservative regime only. PMID- 11019263 TI - Comment on "Structures of GaN(0001)bf-(2x2), bf-(4x4), and bf-(5x5) surface reconstructions" PMID- 11019264 TI - Xue et al. reply: PMID- 11019262 TI - Mixing of atmospheric gas concentrations. AB - Atmospheric gas concentrations were measured at 1 s intervals in the upper troposphere during a flight through and near the anvil of a storm. The observed very high correlations between the concentrations of CO and CH4 are interpreted as arising from the mixing of two distinct air masses with differing concentrations of each species, and is due to the nearly identical diffusivities of CO and CH4 in air. We find that the correlations depend on the period over which each concentration measurement was made. Correlations in measurements made over short periods decay with time, while correlations over larger scales remain high. We interpret this using a simple mixing model. PMID- 11019265 TI - Comment on "Experimental observation of spontaneous emission cancellation" PMID- 11019266 TI - Comment on "Spin dependent hopping and colossal negative magnetoresistance in epitaxial Nd0.52Sr0.48MnO3 films in fields up to 50 T" PMID- 11019267 TI - Wagner et al. reply: PMID- 11019268 TI - Off-diagonal geometric phases. AB - We investigate the adiabatic evolution of a set of nondegenerate eigenstates of a parametrized Hamiltonian. Their relative phase change can be related to geometric measurable quantities that extend the familiar concept of Berry phase to the evolution of more than one state. We present several physical systems where these concepts can be applied, including an experiment on microwave cavities for which off-diagonal phases can be determined from published data. PMID- 11019269 TI - Optical potentials for inelastic scattering from many-body targets. AB - The standard text book Green's function possesses a self-energy that is known to be an optical potential for elastic scattering. The introduction of an optical potential reduces the complex many-body scattering problem into a tractable one body problem. In this paper inelastic Green's functions are introduced and discussed which possess self-energies that are optical potentials for inelastic scattering. If the projectile is indistinguishable from particles comprising the target, intriguing aspects arise even for noninteracting particles. PMID- 11019270 TI - Manifestation of superfluidity in an evolving Bose-Einstein condensed gas. AB - We study the generation of excitations due to an "impurity" (static perturbation) placed into an oscillating Bose-Einstein condensed gas in the time-dependent trapping field. It is shown that there are two regions for the position of the local perturbation. In the first region the condensate flows around the impurity without generation of excitations demonstrating superfluid properties. In the second region the creation of excitations occurs, at least within a limited time interval, revealing destruction of superfluidity. The phenomenon can be studied by measuring the damping of condensate oscillations at different positions of the impurity. PMID- 11019271 TI - Influence of capillary condensation on the near-critical solvation force. AB - We argue that in a fluid, or magnet, confined by adsorbing walls which favor liquid, or the (+) phase, the solvation (Casimir) force in the vicinity of the critical point is strongly influenced by capillary condensation which occurs below the bulk critical temperature T(c). At T slightly below and above T(c), a small bulk field h<0, which favors gas, or the (-) phase, leads to residual condensation and a solvation force which is much more attractive (at the same large wall separation) than that found exactly at the critical point. Our predictions are supported by results obtained from density-matrix renormalization group calculations in a two-dimensional Ising strip subject to identical surface fields. PMID- 11019272 TI - First dark matter limits from a large-mass, low-background, superheated droplet detector. AB - We report on the fabrication aspects and calibration of the first large active mass ( approximately 15 g) modules of SIMPLE, a search for particle dark matter using superheated droplet detectors (SDDs). While still limited by the statistical uncertainty of the small data sample on hand, the first weeks of operation in the new underground laboratory of Rustrel-Pays d'Apt already provide a sensitivity to axially coupled weakly interacting massive particles (WIMPs) competitive with leading experiments, confirming SDDs as a convenient, low-cost alternative for WIMP detection. PMID- 11019273 TI - Exact wave functions in a noncommutative field theory. AB - We consider the nonrelativistic field theory with a quartic interaction on a noncommutative plane and compute the 2-->2 scattering amplitude within perturbative analysis to all orders. We regain the results of the perturbative analysis by finding the scattering and the bound state wave functions of the two particle Schrodinger equation. These wave functions unusually have two center positions in the relative coordinates, whose separation is transverse to the total momentum and scales linearly with its magnitude, exhibiting the stringy nature of the noncommutative field theory. PMID- 11019274 TI - Stability of global monopoles revisited. AB - We analyze the stability of global O(3) monopoles in the infinite cutoff (or scalar mass) limit. We obtain the perturbation equations and prove that the spherically symmetric solution is classically stable (or neutrally stable) to axially symmetric, square integrable, or power-law decay perturbations. Moreover, we show that, in spite of the existence of a conserved topological charge, the energy barrier between the monopole and the vacuum is finite even in the limit where the cutoff is taken to infinity. This feature is specific of global monopoles and independent of the details of the scalar potential. PMID- 11019277 TI - Fully relativistic coupled cluster treatment for parity-violating energy differences in molecules. AB - Fully relativistic four-component many-body perturbation and coupled cluster electronic structure calculations including electroweak neutral current corrections are presented for a study of parity-violating effects in chiral molecules. For the chiral molecules H2O2 and H2S2, it is demonstrated that electron correlation contributions to the parity-violating energy shifts are small but critically dependent on the molecular geometry. PMID- 11019276 TI - Charged-particle multiplicity near midrapidity in central Au+Au collisions at sqrt[SNN]=56 and 130 GeV. AB - We present the first measurement of pseudorapidity densities of primary charged particles near midrapidity in Au+Au collisions at sqrt[s(NN)] = 56 and 130 GeV. For the most central collisions, we find the charged-particle pseudorapidity density to be dN/deta|(|eta|<1) = 408+/-12(stat)+/-30(syst) at 56 GeV and 555+/ 12(stat)+/-35(syst) at 130 GeV, values that are higher than any previously observed in nuclear collisions. Compared to proton-antiproton collisions, our data show an increase in the pseudorapidity density per participant by more than 40% at the higher energy. PMID- 11019279 TI - Mirroring doubly excited resonances in argon. AB - New features are revealed in the low-energy photoionization spectrum of Ar by critically combining high photon resolution and differential photoelectron spectroscopic techniques. Two LS-forbidden doubly excited resonances are seen in the 3p(-1)(3/2, 1/2) partial cross sections which exhibit mirroring profiles, resulting in complete cancellation in the total photoionization cross section, as was predicted by Liu and Starace [Phys. Rev. A 59, R1731 (1999)]. These results demonstrate that a new class of weakly spin-orbit induced, mirroring resonances should be observable in partial, but not in total, collisional cross sections involving atoms, molecules, and solids in general. PMID- 11019278 TI - 1s lamb shift in hydrogenlike uranium measured on cooled, decelerated ion beams. AB - The Lyman- alpha transitions of hydrogenlike uranium associated with electron capture were measured in collisions of stored bare U (92+) ions with gaseous targets at the storage ring ESR. By applying the deceleration technique, the experiment was performed at slow collision energies in order to reduce the uncertainties associated with Doppler corrections. From the measured centroid energies, a ground state Lamb shift of 468 eV+/-13 eV is deduced which gives the most precise test of quantum electrodynamics for a single electron system in the strong field regime. In particular, the technique applied paves the way towards the 1 eV precision regime. PMID- 11019280 TI - Cold collision frequency shifts in a 87Rb atomic fountain. AB - We present measurements of cavity frequency pulling and collisional frequency shifts in a 87Rb fountain with a frequency resolution of 3x10(-16). Agreement with theory is found for the cavity pulling and the measured collisional shifts. The clock shift is found at least 50 times smaller than in 133Cs. PMID- 11019275 TI - Measurements of the mass, total width, and two-photon partial width of the eta(c) meson. AB - Using 13.4 fb(-1) of data collected with the CLEO detector at the Cornell Electron Storage Ring, we have observed 300 events for the two-photon production of ground-state pseudoscalar charmonium in the decay eta(c)-->K(0)(S)K-/+pi(+/-). We have measured the eta(c) mass to be [2980.4+/-2.3 (stat)+/-0.6 (syst)] MeV and its full width as [27.0+/-5.8 (stat)+/-1.4 (syst)] MeV. We have determined the two-photon partial width of the eta(c) meson to be [7.6+/-0.8 (stat)+/-0.4 (syst)+/-2.3 (br)] keV, with the last uncertainty associated with the decay branching fraction. PMID- 11019281 TI - Wave packet echoes in the motion of trapped atoms. AB - We experimentally demonstrate and systematically study the stimulated revival (echo) of motional wave packet oscillations. For this purpose, we prepare wave packets in an optical lattice by nonadiabatically shifting the potential and stimulate their reoccurrence by a second shift after a variable time delay. This technique, analogous to spin echoes, enables one even in the presence of strong dephasing to determine the coherence time of the wave packets. We find that for strongly bound atoms it is comparable to the cooling time and much longer than the inverse of the photon scattering rate. PMID- 11019282 TI - Coherently controlled nanoscale molecular deposition. AB - Quantum interference effects are shown to provide a means of controlling and enhancing the focusing of a collimated neutral molecular beam onto a surface. The nature of the aperiodic pattern formed can be altered by varying laser field characteristics and the system geometry. PMID- 11019283 TI - Dynamical Auger emission induced by multistate vibronic coupling in the core excited states of the BCl3 molecule. AB - The Auger emission following the resonant excitation of the BCl3 molecule has been investigated using monochromatized synchrotron radiation. The B 1s excitation to the unoccupied Anti-bonding 4e(') orbital enhances the shoulder structure in the low kinetic energy side of the photoemission from the 2e(') valence bonding orbital. Based on a series of quantum mechanical calculations, this shoulder structure is interpreted as the dynamical Auger emission which reflects the B-Cl stretching nuclear motion and appears as a result of the purely multistate vibronic coupling effect among the Jahn-Teller split B 1s(-1)4e(') E' states and the closely lying B 1s(-1)3a(')(1) A'1 state. PMID- 11019285 TI - Subsurface-channeling-like energy loss structure of the skipping motion on an ionic crystal. AB - The skipping motion of Ne+ ions in grazing scattering from the LiF(001) surface is studied for velocity below 0.1 a.u. with a time-of-flight technique. It is demonstrated that suppression of electronic excitation and dominance of optical phonon excitation in the projectile stopping results in an odd 1,3,5,... progression of the energy loss peaks, a feature usually ascribed to subsurface channeling. The experimental findings are well reproduced by parameter-free model calculations where thermal vibrations are the dominant cause for the ion trapping and detrapping. PMID- 11019284 TI - Observation of a continuum-continuum interference hole in ultrafast dissociating core-excited molecules. AB - The femtosecond dissociation of HCl after core excitation has been studied through the resonant Auger decay. The spectra contain contributions from decay occurring at both "molecular" and "atomic" internuclear distances. We have observed a new interference mechanism in these spectra: An atomic spectral line develops into a negative spectral contribution, a "hole," when detuning the excitation energy from the maximum of the Cl2p(-1)sigma(*) resonance. Resonant x ray scattering theory quantitatively explains this behavior as due to a novel destructive continuum-continuum interference between molecular and atomic contributions to the Auger decay. PMID- 11019286 TI - Twist mode in spherical alkali metal clusters. AB - A remarkable orbital quadrupole magnetic resonance, so-called twist mode, is predicted in alkali metal clusters where it is represented by Ipi = 2(-) low energy excitations of valence electrons with strong M2 transitions to the ground state. We treat the twist by both macroscopic and microscopic ways. In the latter case, the shell structure of clusters is fully exploited, which is crucial for the considered size region ( 8>tau(d), while it reaches a universal value in the opposite limit. Thus, the Lyapunov exponent of chaotic mesoscopic systems may be found by shot noise measurements. PMID- 11019290 TI - Low frequency fluctuations in a multimode semiconductor laser with optical feedback. AB - We study a multimode semiconductor laser subject to a moderate optical feedback. The steady state is destabilized by either a simple Hopf bifurcation leading to in phase dynamics or by a degenerate Hopf bifurcation leading to antiphase dynamics. The degenerate bifurcation is also a source of multiple coexisting attractors. We show that a simple interpretation of the low frequency fluctuations in the multimode regime is provided by a chaotic itinerancy among the many coexisting unstable attractors produced by the degenerate Hopf bifurcation. PMID- 11019291 TI - Cylindrical microcavity laser based on the evanescent-wave-coupled gain. AB - A microcavity laser based on the gain only in the evanescent field region of whispering gallery modes has been demonstrated. A cylindrical microcavity of 125 microm diam was surrounded by rhodamine 6G dye molecules in an ethanol solution of lower refractive index such that whispering gallery modes of the microcavity underwent laser oscillation when the dye molecules in the evanescent field region outside the cavity were excited by a second harmonic of a Nd:YAG laser. For particular pumping spots, single-mode laser oscillation of a transverse magnetic mode was observed at about 600 nm with associated cavity Q of 3x10(7). PMID- 11019292 TI - Does water vapor prevent upscaling sonoluminescence? AB - Experimental results for single-bubble sonoluminescence of air bubbles at very low frequency f = 7.1 kHz are presented: In contrast to the predictions of a recent model [S. Hilgenfeldt and D. Lohse, Phys. Rev. Lett. 82, 1036 (1999)], the bubbles are only as bright (10(4)-10(5) photons per pulse) and the pulses as long (approximately 150 ps) as at f = 20 kHz. We can theoretically account for this effect by incorporating water vapor into the model: During the rapid bubble collapse a large amount of water vapor is trapped inside the bubble, resulting in an increased heat capacity and hence lower temperatures, i.e., hindering upscaling. At this low frequency water vapor also dominates the light emission process. PMID- 11019293 TI - Chaos healing by separatrix disappearance and quasisingle helicity states of the reversed field pinch. AB - The resilience to chaotic perturbations of one-parameter one-degree-of-freedom Hamiltonian dynamics is shown to increase when its corresponding separatrix vanishes due to a saddle-node bifurcation. This is first highlighted for the magnetic chaos related to quasisingle helicity (QSH) states of the reversed field pinch. It provides a rationale for the confinement improvement of helical structures experimentally found for QSH plasmas; such a feature would not be expected from the classical resonance overlap picture as the separatrix disappearance occurs when the amplitude of the dominant mode increases. PMID- 11019294 TI - Dynamics of electron-plasma vortex in background vorticity distribution. AB - Dynamics of a point vortex in interaction with a broad profile of background vorticity is studied experimentally by using an electron plasma. The observed motion of the vortex compares favorably with a recently proposed theoretical model [D. A. Schecter and D. H. E. Dubin, Phys. Rev. Lett. 83, 2191 (1999)]. Perturbations in the background distribution in the wake of the spiral orbit of the vortex amount to several tens of percent and are considered to be a major reason for deviations of the observation from the linear theoretical model. PMID- 11019295 TI - Chaotic regime of Alfven Eigenmode wave-particle interaction. AB - The chaotic regime in Alfven eigenmode wave-particle interaction is identified for the first time in the tokamak plasma of the Joint European Torus. The Alfven modes are driven by energetic hydrogen minority ions produced by ion cyclotron resonance heating. The experimental signatures of the chaotic regime include spectral broadening, phase flips, and nonperiodic amplitude variations. These phenomena are found to be consistent with a general nonlinear theory of kinetic instabilities near stability threshold developed by Berk, Breizman, and Pekker. PMID- 11019296 TI - Combined X-Ray and fully leaky guided mode studies of the smectic layer and optic tensor configuration in a ferroelectric liquid-crystal cell. AB - X-ray scattering together with optical characterization using fully leaky guided modes have been used for the first time to study the same ferroelectric liquid crystal cell. This enables direct calculation of an accurate cone and chevron description of the liquid-crystal director profile since the layer structure and optic tensor configuration are both well known. PMID- 11019297 TI - Origin of low-frequency local vibrational modes in high density amorphous ice. AB - Incoherent-inelastic neutron scattering data are obtained from 5-80 K for high density amorphous (hda) ice in the region 0-135 cm(-1). An excess contribution to the vibrational density of states is identified near 20 cm(-1). The origin of these vibrations has been identified by lattice dynamics calculations on an "experimental" structure derived from reverse Monte Carlo analysis of hda ice neutron diffraction data. An interpretation that localized oscillations of short chains and isolated water molecules are responsible for the excess low-frequency modes is consistent with our data. PMID- 11019298 TI - Silicon clathrate with an f-electron system. AB - A novel crystal of Ba6Ce2Au4Si42 with Ba and Ce encapsulated into silicon polyhedral clusters is self-assembled from the state of elemental mixture. Each atom in the crystal is arranged in its well-defined position with a nanoscale period, causing unique interactions between the conduction and the magnetic electrons originating from the independent sources of Ba and Ce, respectively. In this system, the long-distant magnetic f electrons can interact with each other through nanoscale spacing with isotropic three dimensionality, leading to the occurrence of a unique spontaneous spin ordering at 6.5 K. PMID- 11019299 TI - Catalytic action of Ni atoms in the formation of carbon nanotubes: a molecular dynamics study. AB - Catalytic action of Ni atoms in the growth of single-wall carbon nanotubes is investigated using tight-binding molecular dynamics and ab initio methods. Our results demonstrate this to be a two step process in which the Ni atom first creates and stabilizes defects in nanotubes. The subsequent incorporation of incoming carbon atoms anneals the Ni-stabilized defects freeing the Ni atom to repeat the catalytic process. PMID- 11019300 TI - Theory of distinct crystal structures of polymerized fullerides AC60, A = K, Rb, Cs: the specific role of alkalis. AB - The polymer phases of AC60 form distinct crystal structures characterized by the mutual orientations of the (C( -)(60))(n) chains. We show that the direct electric quadrupole interaction between chains always favors the orthorhombic structure Pmnn with alternating chain orientations. However, the specific quadrupolar polarizability of the alkali metal ions leads to an indirect interchain coupling which favors the monoclinic structure I2/m with equal chain orientations. The competition between direct and indirect interactions explains the structural difference between KC60 and RbC60, CsC60. PMID- 11019301 TI - Mechanical damping by intercrystalline diffusion of hydrogen in metallic polycrystals. AB - Anelastic relaxation by intercrystalline atomic diffusion-analogous to thermal diffusion (i.e., thermoelastic) effects but never observed hitherto-was recently suggested as a new mechanism to explain part of the hydrogen damping spectra in intermetallic compounds. A critical experimental test of this model is now presented using a Zr65Cu17.5Ni10Al7.5 alloy, which allows for a quantitative kinetic analysis in comparison to a closely related reorientation process. The results are in full agreement with the predictions of the model and clearly corroborate the proposed "intercrystalline Gorsky effect" as a new type of mechanical damping in sufficiently fine-grained polycrystals. PMID- 11019302 TI - Measurement of shock wave rise times in metal thin films. AB - We have measured the rise time of laser-generated shock waves in vapor plated metal thin films using frequency-domain interferometry with subpicosecond time resolution. 10%- 90% rise times of <6.25 ps were found in targets ranging from 0.25 to 2.0 microm in thickness. Particle and average shock velocities were simultaneously determined. Shock velocities of approximately 5 nm/ps were inferred from the measured free surface velocity, corresponding to pressures of 30-50 kbar. Thus, the shock front extends only a few tens of lattice spacings. PMID- 11019303 TI - Phonon dispersion of bcc iron to 10 GPa. AB - The phonon dispersion of bcc iron under high pressure to 10 GPa was measured at 300 K by inelastic neutron scattering. Its pressure dependence is surprisingly uniform. Contrary to the behavior found in other bcc elements, there is a lack of any significant pretransitional behavior close to the martensitic bcc-hcp transition which could be related to the Burgers mechanism. This finding confirms predictions by spin-polarized total energy calculations that explain the transition by the effect of pressure on the magnetism of iron. The high pressure frequencies were used to develop a lattice dynamical model from which thermodynamic quantities can be determined at any pressure to 10 GPa. PMID- 11019304 TI - Minimizing boundary reflections in coupled-domain simulations. AB - We propose a time-dependent boundary condition coupling an atomistic simulation system to linear surroundings such that reflection of elastic waves across the boundary is minimized. Interdomain interactions expressed in terms of memory kernel functions within linear-response theory are treated in a natural dynamical manner, albeit numerically. The approach is shown to give significantly reduced phonon reflections at the domain boundaries relative to existing coupling methods. In addition, we demonstrate that the framework is also effective in the context of static relaxation of displacement fields associated with embedded inhomogeneities. PMID- 11019305 TI - Order-disorder transition in the solid phase of a charged hard sphere model. AB - We investigate the solid phases of the restricted primitive model (RPM). Monte Carlo simulations show the existence of an order-disorder transition from a substitutionally disordered face centered cubic lattice (fcc) to a new ordered fcc structure which is proposed as the ground state of the RPM at the close packing density. Our results suggest that the new phase might turn out in a new triple point in the RPM phase diagram involving three solid phases: CsCl, fcc ordered and fcc disordered structures. The order-disorder transition is also studied using the cell theory. The theory shows good agreement with the simulation results and suggests that the transition is weakly first order. PMID- 11019306 TI - Molecular simulation of ultrathin polymeric films near the glass transition. AB - Properties such as the glass transition temperature ( T(g)) and the diffusion coefficient of ultrathin polymeric films are shown to depend on the dimensions of the system. In this work, a hard-sphere molecular dynamics methodology has been applied to simulate such systems. We investigate the influence that substrates have on the behavior of thin polymer films; we report evidence suggesting that, depending on the strength of substrate-polymer interactions, the glass transition temperature for a thin film can be significantly lower or higher than that of the bulk. PMID- 11019307 TI - Nonstoichiometric interfaces and Al2O3 adhesion with Al and Ag. AB - We have determined the relative stability of stoichiometric, oxygen-rich, and aluminum-rich Al/Al 2O (3) and Ag/Al 2O (3) interfaces from first principles. Stable structures vary significantly with oxygen chemical potentials. Computed works of adhesion agree reasonably well with sessile drop experimental values, including correlation with measured oxygen chemisorption effects on Ag. The ordering of predicted bond energies of the interfaces, ceramics, and metals seems consistent with monotonic and fatigue fracture experiments. PMID- 11019309 TI - Long-range correlations in the nonequilibrium quantum relaxation of a spin chain. AB - We consider the nonstationary quantum relaxation of the Ising spin chain in a transverse field of strength h. Starting from a homogeneously magnetized initial state the system approaches a stationary state by a process possessing quasi-long range correlations in time and space, independent of the value of h. In particular, the system exhibits aging (or lack of time-translational invariance on intermediate time scales) although no indications of coarsening are present. PMID- 11019308 TI - Kinetic roughening in polymer film growth by vapor deposition. AB - The growth front roughness of linear poly( p-xylylene) films grown by vapor deposition polymerization has been investigated using atomic force microscopy. The interface width w increases as a power law of film thickness d, w approximately d(beta), with beta = 0. 25+/-0.03, and the lateral correlation length xi grows as xi approximately d(1/z), with 1/z = 0.31+/-0.02. This novel scaling behavior is interpreted as the result of monomer bulk diffusion, and belongs to a new universality class that has not been discussed previously. PMID- 11019310 TI - Stability-instability transitions in silicon crystal growth. AB - In order for a crystal to grow, source atoms must be incorporated into the underlying lattice. Typically, this process occurs on the surface in one of two modes: either through island nucleation or through step flow. However, a third, morphologically unstable growth mode has been predicted. Monitoring the surface of ultraflat substrates with an in situ scanning electron microscope, we prove that for the (111) face of silicon there is a transition from stable step flow to morphological instability and then to island nucleation. PMID- 11019311 TI - Hole dynamics in noble metals. AB - Hole dynamics in noble metals (Cu and Au) is investigated by means of first principles many-body calculations. While holes in a free-electron gas are known to live shorter than electrons with the same excitation energy, our results indicate that d holes in noble metals exhibit longer inelastic lifetimes than excited sp electrons, in agreement with experiment. The density of states available for d-hole decay is larger than that for the decay of excited electrons; however, the small overlap between d and sp states below the Fermi level increases the d-hole lifetime. The impact of d-hole dynamics on electron hole correlation effects is also addressed. PMID- 11019312 TI - Hydrogen bonding and dipole moment of water at supercritical conditions: a first principles molecular dynamics study. AB - We present a first-principles molecular dynamics study of water near and above the critical point ( T = 647 K, rho = 0.32 g/cm(3)). We find that the systems undergo fast dynamics with continuous formation and breaking of H bonds. At low density, the system fragments mostly into trimers, dimers, and single molecules. At a higher density, more complex structures appear and an extended, albeit very dynamical, H-bond network can be identified. These structures have important consequences for the screening properties of the system. This offers a clue to understanding the peculiar chemical behavior of a supercritical system and allows thermodynamical tuning of its solvent properties. PMID- 11019313 TI - Stability of carbon nanotubes: how small can they be? AB - Experimental evidence has been found for the existence of small single wall carbon nanotubes with diameters of 0.5 and 0.33 nm by high resolution transmission electron microscopy, and their mechanical stability was investigated using tight-binding molecular dynamics simulations. It is shown that, while the carbon tubes with diameters smaller than 0.4 nm are energetically less favorable than a graphene sheet, some of them are indeed mechanically stable at temperatures as high as 1100 degrees C. The 0.33 nm carbon tube observed is likely a (4, 0) tube and is indeed part of a compound nanotube system that forms perhaps the smallest metal-semiconductor-metal tubular junction yet synthesized. PMID- 11019314 TI - Current mirror effect and correlated cooper-pair transport in coupled arrays of small Josephson junctions. AB - We have measured the transport properties of capacitively coupled one-dimensional arrays of small aluminum Josephson junctions. Under suitable biasing conditions, the samples act as current mirrors; the currents in the two arrays couple to each other and become equally large. The coupling is found both for currents flowing in the same direction and in opposite directions. We have also measured the time dependent fluctuations of the currents in the two arrays, and we find a strong correlation or anticorrelation of the fluctuations depending on the relative direction of the two currents. PMID- 11019316 TI - Neutron resonance: modeling photoemission and tunneling data in the superconducting state of Bi2Sr2CaCu2O8+delta. AB - Motivated by neutron scattering data, we develop a model of electrons interacting with a magnetic resonance and use it to analyze angle resolved photoemission and tunneling data in the superconducting state of Bi(2)Sr(2)CaCu(2)O(8+delta). We not only can explain the peak-dip-hump structure observed near the (pi,0) point, and its particle-hole asymmetry as seen in superconductor-insulator-normal tunneling spectra, but also its evolution throughout the Brillouin zone, including a velocity "kink" near the d-wave node. PMID- 11019315 TI - Reorientation of anisotropy in a square well quantum hall sample. AB - We have measured magnetotransport at half-filled high Landau levels in a quantum well with two occupied electric subbands. We find resistivities that are isotropic in perpendicular magnetic field but become strongly anisotropic at nu = 9/2 and 11/2 on tilting the field. The anisotropy appears at an in-plane field, B(ip) approximately 2.5 T, with the easy-current direction parallel to B(ip) but rotates by 90 degrees at B(ip) approximately 10 T and points now in the same direction as in single-subband samples. This complex behavior is in quantitative agreement with theoretical calculations based on a unidirectional charge density wave state model. PMID- 11019317 TI - Equilibration and dynamic phase transitions of a driven vortex lattice. AB - We report on the observation of two types of current driven transitions in metastable vortex lattices. The metastable states, which are missed in usual slow transport measurements, are detected with a fast transport technique in the vortex lattice of undoped 2H-NbSe2. The transitions are seen by following the evolution of these states when driven by a current. At low currents we observe an equilibration transition from a metastable to a stable state, followed by a dynamic crystallization transition at high currents. PMID- 11019318 TI - Field induced ordering in highly frustrated antiferromagnets. AB - We predict that an external field can induce a spin ordering in highly frustrated classical Heisenberg magnets. We find analytically stabilization of collinear states by thermal fluctuations at a one-third of the saturation field for kagome and garnet lattices and at a half of the saturation field for pyrochlore and frustrated square lattices. This effect is studied numerically for the frustrated square-lattice antiferromagnet by Monte Carlo simulations for classical spins and by exact diagonalization for S = 1/2. The field induced collinear states have a spin gap and produce magnetization plateaus. PMID- 11019319 TI - Magnetism in Al(Si)-Mn quasicrystals and related phases. AB - An extreme sensitivity of the magnetic properties to the atomic structure has been observed in Al(Si)-Mn and Al-Pd-Mn quasicrystals, approximants, and liquids with similar Mn concentrations (of the order of 8%- 20%). Here, we show that the effect of the local environment of the Mn atoms is not sufficient to explain this complex behavior. A new model is presented, which, taking into account Mn-Mn interactions mediated by the conduction electrons over large distances ( 5 A and more), allows one to understand why only a small fraction of the Mn atoms carry a localized magnetic moment in quasicrystals and why a large proportion is magnetic in liquids. PMID- 11019320 TI - Thermal spin-wave scattering in hot-electron magnetotransport across a spin valve. AB - The role of thermal scattering in spin-dependent transport of hot electrons at 0.9 eV is studied using a spin-valve transistor with a soft Ni(80)Fe(20)/Au/Co base. Spin-dependent scattering makes the collected electron current depend sensitively on the magnetic state of the base. The magnetocurrent reaches 560% at 100 K, decays with increasing temperature, and a huge effect of 350% still remains at room temperature. The results demonstrate that thermal spin waves produce quasielastic spin-flip scattering of hot electrons, resulting in mixing of the two spin channels. PMID- 11019321 TI - Spin-wave interaction and renormalization of magnetic anisotropy in 2D antiferromagnets. AB - The renormalization of the magnetic anisotropy is investigated in 2D easy-axis antiferromagnets using 1/S expansion. It was shown that in the spin-wave gap Delta appears as the 1/S2 infrared divergent correction which becomes large at T>Delta and demands analysis of the full 1/S expansion. Qualitative consequences of this gap renormalization are discussed in relation to the AFR and neutron scattering experiments. PMID- 11019322 TI - Observation of two time scales in the ferromagnetic manganite La1-xCaxMnO3, x approximately 0.3. AB - We report new zero-field muon spin relaxation and neutron spin echo measurements in ferromagnetic (FM) (La,Ca)MnO3 which suggest at least two spatially separated regions possessing very different Mn-ion spin dynamics. One region displays diffusive relaxation, "critical slowing down" near T(C) and an increasing volume fraction below T(C), suggesting overdamped FM spin waves below T(C). The second region possesses more slowly fluctuating spins, a linewidth independent of q, and a decreasing volume fraction below T(C). The estimated length scale for the inhomogeneity is 0.5). AB - We report an inelastic light scattering study of the effects of charge ordering on the spin, charge, and lattice dynamics of Bi1-xCaxMnO3 (x>0.5). We find that charge ordering results in anomalous phonon behavior, such as the appearance of "activated" modes. More significantly, however, the transition to the charge ordered phase results in the appearance of a quasielastic scattering response with the symmetry of the spin-chirality operator ( T(1g)); this scattering response is thus indicative of magnetic or chiral spin fluctuations in the antiferromagnetic charge-ordered phase. PMID- 11019326 TI - Correlation between fluorescence intermittency and spectral diffusion in single semiconductor quantum dots. AB - We find a correlation between the dynamics of fluorescence intermittency and spectral diffusion in the spectroscopy of single CdSe nanocrystal quantum dots (QD). A statistical analysis of the data suggests two populations of blinking events: blinking followed by large spectral diffusion shifts and blinking with small or no spectral shifts. Although unexpected from earlier studies, the correlation between blinking and spectral shifting is consistent with a model of QD ionization as the mechanism for the blinking event, followed by a redistribution of local electric fields that results in spectral shifting. PMID- 11019327 TI - Mechanism of photoinduced changes in the structure and optical properties of amorphous As2S3. AB - We propose a mechanism of photostructural changes in amorphous As2S3 ( a-As2S3) on the basis of ab initio molecular orbital calculations on clusters of atoms modeling the local structure of the amorphous system. We have found that trigonal AsS3 pyramidal units can be transformed into a fivefold coordinated As site having four As-S bonds and one As-As bond via a photoionization process. This photoinduced coordination defect center exhibits a lower photoabsorption energy as compared with the usual pyramidal structure, explaining the observed photodarkening effect of a-As2S3. PMID- 11019328 TI - How carbon monoxide adsorbs in different sites. AB - The interplay between the electronic and the geometric structure of adsorbates is of fundamental importance for the understanding of many surface phenomena. Using x-ray emission spectroscopy and ab initio cluster calculations, this issue has been investigated in unprecedented detail for CO adsorption in different adsorption sites. The investigation establishes pi bonding and sigma repulsion, both increasing with the number of coordinated metal atoms. The two contributions partly compensate each other, leading to only small differences in adsorption energies for the different adsorption sites despite very large variations in the electronic structure. PMID- 11019329 TI - Quantum cryptography with 3-state systems. AB - We consider quantum cryptographic schemes where the carriers of information are 3 state particles. One protocol uses four mutually unbiased bases and appears to provide better security than obtainable with 2-state carriers. Another possible method allows quantum states to belong to more than one basis. Security is not better, but many curious features arise. PMID- 11019330 TI - Hydrodynamic coupling of two brownian spheres to a planar surface. AB - We describe direct imaging measurements of the collective and relative diffusion of two colloidal spheres near a flat plate. The bounding surface modifies the spheres' dynamics, even at separations of tens of radii. This behavior is captured by a stokeslet analysis of fluid flow driven by the spheres' and wall's no-slip boundary conditions. In particular, this analysis reveals surprising asymmetry in the normal modes for pair diffusion near a flat surface. PMID- 11019331 TI - Disorder induced diffusive transport in ratchets. AB - The effects of quenched disorder on the overdamped motion of a driven particle on a periodic, asymmetric potential are studied. While for the unperturbed potential the transport is due to a regular drift, the quenched disorder induces a significant additional chaotic "diffusive" motion. Possible applications to experiments in nanoscale surfaces and particle separation are discussed. PMID- 11019332 TI - Multifractal wavelet filter of natural images. AB - Natural images are characterized by the multiscaling properties of their contrast gradient, in addition to their power spectrum. In this Letter we show that those properties uniquely define an intrinsic wavelet and present a suitable technique to obtain it from an ensemble of images. Once this wavelet is known, images can be represented as expansions in the associated wavelet basis. The resulting code has the remarkable properties that it separates independent features at different resolution level, reducing the redundancy, and remains essentially unchanged under changes in the power spectrum. The possible generalization of this representation to other systems is discussed. PMID- 11019333 TI - Anharmonic effects on charge and orbital order. PMID- 11019334 TI - Comment on "Kagome lattice antiferromagnet stripped to its basics". PMID- 11019336 TI - Comment on "Triviality of the ground state structure in Ising spin glasses". PMID- 11019338 TI - Acquired hypogonadotropic hypogonadism presenting as decreased seminal volume. AB - A 32-year-old man with decreased ejaculatory volume was found to have acquired hypogonadotropic hypogonadism. Initial evaluation demonstrated castrate levels of testosterone with low serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels. Semen analysis revealed a volume of 0.35 cc and severe oligospermia. Administration of gonadotropin-releasing hormone (GnRH) did not effect an increase in LH or FSH, indicating a pituitary defect. Magnetic resonance imaging revealed a partially empty sella turcica. Treatment with human chorionic gonadotropin (hCG) alone resulted in normalization of testosterone levels, sperm concentration, and semen volume, as well as the successful conception and delivery of a healthy baby girl. The findings from this case demonstrate the importance of considering low serum testosterone levels in the evaluation of low semen volume, as well as the role of hCG alone as an infertility treatment for acquired hypogonadotropic hypogonadism. PMID- 11019339 TI - Children with feeding disorders: an emerging issue. PMID- 11019340 TI - Virtual estimation: Internet program helps users determine nutrient intake and needs. PMID- 11019341 TI - Down syndrome and folic acid update. PMID- 11019342 TI - Malnutrition awareness activity. PMID- 11019343 TI - Milk consumption for the lactose intolerant: a clarification. PMID- 11019344 TI - Milk consumption for the lactose intolerant: a clarification. PMID- 11019346 TI - ADA leads way in putting children back on the road to good health. PMID- 11019345 TI - Computers and dietetics education. PMID- 11019347 TI - A report from the office of the Surgeon General. PMID- 11019348 TI - Impact of a randomized, controlled trial of liberal vs conservative hospital discharge criteria on energy, protein, and fluid intake in patients who received marrow transplants. AB - OBJECT: To determine if adult patients who received marrow transplants had faster resumption of oral energy and nutrient intake and shorter duration of intravenous (i.v.) fluid requirement if discharged from the hospital earlier than is customary. DESIGN: Randomized, controlled trial of patients remaining hospitalized because of inadequate oral intake. Consenting patients were assigned randomly to remain hospitalized (hospital group) or be discharged to an ambulatory setting (ambulatory group). SUBJECTS: Seventy-eight patients of the Fred Hutchinson Cancer Research Center who were consuming less than 33% of estimated energy requirement and requiring up to 3,000 mL of fluids per day intravenously. INTERVENTION: Participants received nutrition counseling by a registered dietitian to promote resumption of oral intake. Daily oral intake records were analyzed to determine energy and nutrient content. MAIN OUTCOME MEASURES: Days after study enrollment to consume 33% of energy and protein requirements and total number of days of i.v. fluid support were analyzed by group until discharge from the center, approximately 100 days after transplantation. STATISTICAL ANALYSES: Demographic data were defined by group means. Differences between treatment procedures were determined by Cox regression analysis. No variables were confounding. RESULTS: The hospital group took fewer days than the ambulatory group to resume oral energy intake (4.5 vs 8.0, P = .004) and to discontinue i.v. fluids (30.5 vs 48.5, P = .019). There was no difference between groups in days of parenteral nutrition support (P = .817) or days to resume oral protein intake (P = .470). APPLICATIONS/CONCLUSIONS: Oral and gastrointestinal complications delay resumption of oral energy and protein intakes after transplantation. Earlier hospital discharge can achieve cost savings but may delay resumption of oral energy intake. Because of continued high risk nutrition status and potential for rapid change in medical status, nutrition assessment and counseling are necessary in both the hospital and ambulatory setting to promote resumption of oral intake and discontinuation of i.v. fluids. PMID- 11019349 TI - A qualitative study of participants' experiences with dietary assessment. AB - OBJECTIVE: This article reports on the use of focus groups and an experimental participatory activity to investigate factors influencing participants' decisions about what to eat and what to report on food records and food frequency questionnaires. DESIGN: Four focus groups examined participants' experience with diet records and 3 focus groups explored the topic of food portions using a group consensus activity. Twenty-two women participated in the diet record focus groups, and 15 participated in portion estimation groups. SUBJECTS: Focus group participants were equally distributed by age and body mass index values. Each woman completed a 10-day doubly labeled water protocol to measure total energy expenditure, 7 days of diet records (before and during total energy expenditure), and a food frequency questionnaire after the total energy expenditure. ANALYSIS: Transcripts of the focus groups were coded to index categories of responses and to identify themes within and across those responses. Themes discussed in this article are those that were discussed most often and at greatest length by all groups. RESULTS: The diet record focus groups revealed that 2 major factors influenced reporting on diet records: honesty vs social acceptability, and simplifying food intake. The portion estimation focus groups revealed 5 factors that influenced perceptions of portion size: the role of food in the meal, the type of food, personal preferences, product serving sizes, and comparison of personal servings with those of others. APPLICATIONS: The validity and reliability of self-reported food consumption is greatly influenced by the ways people interpret and respond to dietary assessment instruments. These findings indicate that dietetics professionals need to take extra steps to address issues of accurately recording "bad" foods when training patients to complete diet records. Extra probing is needed when dietary records do not include snacks and include simple meals and a large amount of prepared and packaged food because this may indicate that changes in normal dietary patterns were made in order to more easily complete a dietary record. PMID- 11019351 TI - Clinical nutrition managers have access to sources of empowerment. AB - OBJECTIVE: To ascertain perceived access of dietitians to power in the workplace. DESIGN: The conceptual framework was Kanter's theory of organizational power. The Conditions for Work Effectiveness Questionnaire was used to measure perceived access to sources of power: information, support, resources, and opportunities. Demographic data were collected to identify factors that may enhance empowerment. SUBJECTS/SETTINGS: The questionnaire was sent to a random sample of 348 dietitians chosen from members of the Clinical Nutrition Management dietetic practice group of the American Dietetic Association. Blank questionnaires were returned by 99 (28.4%) people not working as clinical nutrition managers, which left 249 in the sample. STATISTICAL ANALYSES: Descriptive statistics were used to organize and summarize data. One-way analysis of variance and t tests were performed to identify differences in responses based on levels of education, work setting, and information technology skills. RESULTS: Usable questionnaires were received from 178 people (71.5%). On a 5-point scale, scores for access to information (mean +/- standard deviation [SD] = 3.8 +/- 0.7), opportunity (mean +/- SD = 3.6 +/- 0.7), support (mean +/- SD = 3.2 +/- 0.9), and resources (mean +/- SD = 3.1 +/- 0.8) demonstrated that clinical nutrition managers perceived themselves as having substantial access to sources of empowerment. Those having higher levels of education, working in larger hospitals, having better-developed information technology skills, and using information technology more frequently had statistically significant higher empowerment scores (P = < or = .05) than contrasting groups in each category. APPLICATIONS/CONCLUSION: Clinical nutrition managers are empowered and able to assume leadership roles in today's health care settings. Their power may be enhanced by asserting more pressure to gain greater access to sources of power: support, information, resources, and opportunities. PMID- 11019350 TI - Role of registered dietitians in dysphagia screening. AB - OBJECTIVE: To examine the ability of registered dietitians to identify patients at risk for dysphagia and make appropriate diet/feeding recommendations in comparison with the speech-language pathologist, and to determine screening criteria for the registered dietitian to use for prediction of dysphagia risk. DESIGN: The dietitian and speech-language pathologist performed dysphagia screening on subjects independently through questioning and/or mealtime observation to identify signs and symptoms of dysphagia. Presence of dysphagia risk and diet/feeding recommendations were determined and results from the dietitian and speech-language pathologist were compared. SUBJECTS/SETTING: Thirty four patients admitted during a 2-month period to a neuroscience unit at an urban teaching hospital were analyzed prospectively. STATISTICAL ANALYSES PERFORMED: kappa Statistics were used to assess agreement between the dietitian and speech language pathologist. A kappa level of less than 0.4 indicated weak agreement, 0.4 to 0.7 indicated moderate agreement, and greater than 0.7 indicated strong agreement. Logistic regression methods were used to evaluate screening criteria as potential predictors of dysphagia risk. RESULTS: Moderate agreement (0.61) was found between the dietitian and speech-language pathologist in determination of dysphagia risk. The dietitian predicted the ability of the patient to consume an oral diet with strong agreement with the speech-language pathologist (1.0); various diet consistencies with moderate agreement (0.61); and the need for liquid restrictions with strong agreement (1.0). The most significant screening variables for prediction of dysphagia risk (P < .05) were age (P = .018), history of dysphagia (P = .042), difficulty swallowing solids (P = .0007), observed facial weakness (P < .0001), and a change in voice quality (P = .0007). Self reported screening variables significantly related to dysphagia risk included drooling of liquids (P = .0009) and solids (P = .0080), facial weakness (P = .0006), change in voice quality (P = .0010), and prolonged eating time (P = .0157). APPLICATIONS/CONCLUSIONS: Dietitians can effectively identify patients with dysphagia. Screening for dysphagia can be implemented as part of standard nutrition assessments and may aid in decreasing dysphagia-related complications. PMID- 11019352 TI - Clinical nutrition and foodservice personnel in teaching hospitals have different perceptions of total quality management performance. AB - OBJECTIVE: To investigate the perceived total quality management (TQM) performance of their department by clinical nutrition managers and dietitians, and foodservice managers and supervisors, in hospital food and nutrition service departments. DESIGN: Using a 2-part questionnaire containing items about 3 constructs of TQM performance and demographic characteristics, participants rated their perceptions of TQM performance. SUBJECTS: Employees in 7 Council of Teaching Hospitals. Of the 128 possible respondents, 73 (57%) completed the study. STATISTICAL ANALYSES PERFORMED: Correlation analysis to identify relationships between demographic characteristics and TQM performance. Analysis of variance to investigate statistical differences among hospitals and between subject groups and types of employment positions. RESULTS: Three TQM constructs- organization, information, and quality management--were evaluated. The clinical nutrition manager and dietitian group had mean ratings between 3.1 and 4.7 (5 point Likert scale); the foodservice manager and supervisor group had mean ratings from 2.7 to 4.0. Education level was significantly correlated (r = 0.44) to performance of employee training in the clinical nutrition group. The number of employees directly supervised was negatively correlated (r = -0.21) to the performance of employee training in the foodservice group. APPLICATIONS: As the dynamic roles of dietitians change, many dietitians will occupy management positions in organizations such as restaurants, health food stores, food processing/distribution companies, and schools. This study demonstrates how a TQM survey instrument could be applied to clinical nutrition and foodservice settings. Dietitians will need to assess TQM in their workplace facilities, especially because of the direct links of TQM to productivity and client satisfaction. PMID- 11019353 TI - Development of a modified picture-sort food frequency questionnaire administered to low-income, overweight, African-American adolescent girls. AB - There is essentially no ideal method of dietary assessment. Physiologic methods (i.e., doubly labeled water) probably come closest, but high cost, participant burden, and limited information limit their use. Furthermore, most dietary assessment methods have been designed for and tested in white adults. Very few have been designed for and tested in African-American adolescents. This study examined validity and reliability of a modified picture-sort food frequency questionnaire (FFQ) administered to 22 low-income, overweight, African-American adolescent girls, aged 11 to 17 years. The FFQ was administered to subjects twice during a 2-week period, and evaluated using the mean values of three 24-hour recalls. The natural log-transformed energy-adjusted, deattenuated correlation coefficients between the second FFQ and the mean from 3 recalls exceeded 0.50 for most nutrients, ranging from 0.32 (protein) to 0.87 (saturated fat). The energy and nutrient values from the first FFQ were greater than those from the second FFQ. Most correlation coefficients for the test-retest reliability of the FFQ were not significant. We conclude that although larger samples are needed to generalize results, the picture-sort dietary assessment method appears to be promising and merits further research. PMID- 11019354 TI - Status of nutrition labeling, health claims, and nutrient content claims for processed foods: 1997 Food Label and Package Survey. AB - The Food and Drug Administration (FDA) conducts studies of food labels as part of its ongoing monitoring of the nutritional status of the US population. In 1994 FDA nutrition labeling rules were implemented and in 1997 the Food Label and Package Survey characterized various aspects of the labeling of processed, packaged foods, including nutrition labeling, health claims, and nutrient content claims. For the survey, FDA selected a multistage, representative sample of food products from the SCAN-TRACK food sales database (AC Nielsen Co, Schaumburg, Ill). FDA identified 58 product groups and selected those product classes from the database that accounted for 80% of sales in each group. From each product class, FDA selected the 3 top-selling product brands and randomly selected follower brands. Based on label information from a final sample of 1,267 food products, FDA determined the percentage of products sold that bear Nutrition Facts labels, health claims, and nutrient content claims. The purpose of this article was to present FDA findings regarding the status of food labels 3 years after implementation of the nutrition labeling rules. Nutrition-labeled products accounted for an estimated 96.5% of the annual sales of processed, packaged foods. An additional 3.4% of products sold were exempt from labeling regulations. Nutrient content claims and health claims appeared on an estimated 39% and 4%, respectively, of the products sold. Dietitians and other health care professionals can use this survey information to identify food types with specific label information and to assist the US consumer in making more varied and healthful food choices in the marketplace. PMID- 11019355 TI - A food display assignment and handling food models improves accuracy of college students' estimates of food portions. PMID- 11019356 TI - Dietary intakes and plasma concentrations of vitamin C are lowered in healthy people with chronic, nonprogressive physical disabilities. PMID- 11019357 TI - High levels of retinol intake during the first trimester of pregnancy result from use of over-the-counter vitamin/mineral supplements. PMID- 11019358 TI - Breast-feeding practices among adolescent and adult mothers in the Missouri WIC population. PMID- 11019359 TI - Apparent decreased energy requirements in children with organic acidemias: preliminary observations. PMID- 11019360 TI - Methods of aggregation of variables in population dynamics. AB - In ecology, we are faced with modelling complex systems involving many variables corresponding to interacting populations structured in different compartmental classes, ages and spatial patches. Models that incorporate such a variety of aspects would lead to systems of equations with many variables and parameters. Mathematical analysis of these models would, in general, be impossible. In many real cases, the dynamics of the system corresponds to two or more time scales. For example, individual decisions can be rapid in comparison to growth of the populations. In that case, it is possible to perform aggregation methods that allow one to build a reduced model that governs the dynamics of a lower dimensional system, at a slow time scale. In this article, we present a review of aggregation methods for time continuous systems as well as for discrete models. We also present applications in population dynamics. A first example concerns a continuous time model of a single population distributed on a system of two connected patches (a logistic source and a sink), by fast migration. It is shown that under a certain condition, the total equilibrium population can be larger than the carrying capacity of the logistic source. A second example concerns a discrete model of a population distributed on two patches, still a source and a sink, connected by fast migration. The use of aggregation methods permits us to conclude that density-dependent migration can stabilize the total population. PMID- 11019362 TI - Is plant biotin holocarboxylase synthetase a bifunctional enzyme? AB - Holocarboxylase synthetases (HCSs) catalyse the biotinylation of biotin-dependent carboxylases in both prokaryotes and eukaryotes. In Escherichia coli and Bacillus subtilis, the protein also acts as a transcriptional repressor that regulates the synthesis of biotin. Previously, we isolated and characterized a cDNA encoding an Arabidopsis thaliana HCS and subsequently assigned this enzyme form to the chloroplast compartment. To investigate whether or not the Arabidopsis protein may function as a regulator in E. coli, we have expressed the functional plant HCS in a birA-derepressed mutant strain of E. coli devoid of the corresponding E. coli protein and carrying a promoter-less LacZ gene marker inserted into the biotin operon, such that the bio promoter drives the synthesis of beta galactosidase. Our data demonstrate that although the expressed plant HCS efficiently complemented the function of apo-carboxylase biotinylation in E. coli, it proved unable to regulate the expression of the biotin biosynthetic genes. PMID- 11019361 TI - [HLA-G: fetomaternal tolerance]. AB - HLA-G is a non-classical major histocompatibility complex class I molecule selectively expressed on cytotrophoblasts. We have demonstrated ex vivo (from voluntary pregnancy interruption samples) the protector role of the HLA-G molecule present on the surface of cytotrophoblast cells versus the lysis carried out by the decidual uterine NK cells. This occurs under semi-allogenic conditions (maternal uterine NK cells and their trophoblast counterparts), as well as in allogenic conditions (maternal uterine NK cells and trophoblast cells from different mothers), thus defining the absence of maternal rejection at the moment of the implantation of the fertilized egg during pregnancy. Moreover, the expression of HLA-G on the cytotrophoblasts permits migration in maternal circulation and infiltration of maternal tissue (particularly in the skin), thereby probably creating a general state of tolerance. In the context of heart transplantation, in preliminary studies, we show that the presence of HLA-G in cardiac biopsy tissue prelevated from grafted patients significantly reduces acute rejects and shows an absence of chronic rejects. In the tumour context, the expression of HLA-G protein at the surface of primitive melanoma and metastatic cells confers protection from NK and CTL lytic activity. This suggests that HLA-G expression may impede the elimination of malignant cells by anti-tumour immune effector cells, constituting a newly described mechanism by which tumour cells may evade immunosurveillance. From there on E.D. Carosella introduced the breakthrough concept of 'HLA a tolerance molecule' in the heart of histocompatibility antigens, which had been described up till then as antigenes of defence and rejection, and the dramatic role of HLA-G in immunotolerance. PMID- 11019363 TI - [Seasonal reproduction of Porcellio variabilis Lucas, 1846]. AB - Porcellio variabilis is common and very widespread in Tunisia. Its breeding period lasts from February to October with a cessation during the months of November, December and January. The recruitment of young takes place principally between April and July. Virgin and mated females reared with natural photoperiod in the laboratory show that the initiation of reproduction does not depend on mating; however, it is accelerated when the male is present, the preparturial moult being shorter in females reared in the presence of a male. PMID- 11019364 TI - [Presence if a new oyster predator, Ocinebrellus inornatus (Recluz, 1851), in the shellfish culture bay of Marennes-Oleron]. AB - The muricid gastropod, Ocinebrellus inornatus, originates from the coasts of the Korean Sea and southern Japan. This species has been regularly sampled in the bay of Marennes-Oleron (France) since spring 1997, and allowed us to validate the taxonomic status of a previous report of the species in the bay in April 1995. O. inornatus was sampled only in the bay of Marennes-Oleron and has not been observed in the other areas along the Charente-Maritime coast. O. inornatus lives mainly in the same biotope as the local muricid Ocenebra erinacea (i.e. the level of Fucus serratus between MLWN and ELWS) Now, this alien species seems to be very well settled in the bay of Marennes-Oleron, where it causes damage to the farming oyster beds. In the areas of highest densities of Ocinebrellus inornatus, the local species Ocenebra erinacea is observed in comparatively reduced numbers. The probable causes of introduction of O. inornatus in the bay are discussed. PMID- 11019365 TI - Myrmecophily in Hesperiidae. The case of Vettius tertianus in ant gardens. AB - The larvae of the hesperiid butterfly Vettius tertianus develop by eating the leaves of Aechmea mertensii, a bromeliad epiphyte restricted to ant gardens. The relationships between ants and V. tertianus larvae highlight the preferential association of the caterpillars with Pachycondyla goeldii (Ponerinae), an ant garden initiator. The oviposition strategy of V. tertianus may thus imply the identification of the inhabiting ant species and not only the identification of the host plant. The caterpillars neither provide secretions to the ants, nor possess defensive devices (i.e. hairs or appendices) against ants. Their activity rhythm does not isolate them from foraging workers of P. goeldii and their shelters are also attainable by the ants. Moreover, as the cuticular lipid profiles of V. tertianus larvae are clearly different from those of the ants and also from the leaf-surface of A. mertensii, acceptance is not due to mimicry between larvae and plants or ants. However, the caterpillars deposit, on the leaf they eat, silk containing a mixture of substances very similar to those found on their own cuticle. No interaction with ants was recorded during observations, even though the ant gardens were patrolled by numerous P. goeldii individuals during their activity period. But when confronted with the caterpillar, none of the tested ant species reacted aggressively. These results suggest the existence of compounds, other than cuticular lipids, responsible for the absence of aggressiveness in the ants. The case of V. tertianus is relatively new as myrmecophily within Hesperiidae has been described only once. Moreover, it preferentially involves a member of the Ponerinae, a subfamily in which interactions with other arthropods are exceptional. PMID- 11019366 TI - [Spawn and spat collecting relationship in Mimachlamys gloriosa (Reeve, 1853) and Brachtechlamys vexillum (Reeve, 1853), Pectinids of New Caledonia]. AB - Pectinidae spat collecting was carried out during a period of 1.5 years in the southern lagoon of New Caledonia. The majority of spat collected were constituted by Mimachlamys gloriosa and Brachtechlamys vexillum larvae. Settlement showed quantitative variations during the year with a maximum in June-September and a minimum (close to zero) in summer (February). However, although adult stocks maintain a stable density and spawn all the year round, spawning intensity shows a maximum in austral summer, contrarily to the recruitment. Therefore, spawning and spat collecting present no correlation and high values of the gonadic index are not followed by numerous spat collecting. These two tropical species present an opportunistic reproductive model. PMID- 11019367 TI - [Cortex-wall connections in the apical cell of Sphacelaria]. AB - The apical cell of Sphacelaria (Fucophyceae) exhibits a permanent polarized organization throughout asymmetric divisions. The apex organization was studied by immunolocalization of tubulin, vitronectin, alpha-actinin and beta 1 integrin. Microfilaments were stained directly by fluorescein phalloidin. The apex was highly organized around a patch of microfilaments densely packed at the tip, where vitronectin-like and alpha-actinin-like proteins colocalized. In the same area, an actin-dependent targeted secretion of sulfated polysaccharides was shown. The permanent localization of these components throughout cell elongation suggests that a cortical site involving transmembrane connections between the cytoskeleton and the extracellular matrix is required for cell polarity. A model of the organization of the tip is proposed. PMID- 11019369 TI - Is the brown shrimp Crangon crangon (L.) population of the Vaccares lagoon (Camargue, France, Rhone delta) an annual population? AB - To study the population dynamics of Crangon crangon in the Vaccares lagoon in the Camargue (Rhone delta), the age of brown shrimp was determined using the method of Tiews (1954). This method consists in counting the number of segments of the outer branch of the first antenna. This number is correlated with the number of moults already accomplished by the shrimp. The duration of the intermoult depends on water temperature. Adapting this model to our environmental data, we determined the age of the brown shrimp. Hatching of larvae occurs in the sea in May, and juveniles enter the lagoon from June to September. Our results showed that Crangon crangon in the Vaccares lagoon reached sexual maturity between 6 and 11 months, and individuals are 1 year old when they leave the lagoon. A few adults from the previous year (1+) migrate back to the lagoon the following year, but these older individuals represent only a very small part of the population, and have only been observed occasionally. The Vaccares brown shrimp population can therefore be considered as annual, in contrast to northern European populations. PMID- 11019368 TI - The NO way to increase muscular utrophin expression? AB - Duchenne muscular dystrophy (DMD), a severe X-linked recessive disorder that results in progressive muscle degeneration, is due to a lack of dystrophin, a membrane cytoskeletal protein. An approach to the search for a treatment is to compensate for dystrophin loss by utrophin, another cytoskeletal protein. During development, in normal as in dystrophic embryos, utrophin is found at the membrane surface of immature skeletal fibres and is progressively replaced by dystrophin. Thus, it is possible to consider utrophin as a 'foetal homologue' of dystrophin. In a previous work, we studied the effect of L-arginine, the substrate of nitric oxide synthetase (NOS), on utrophin expression at the muscle membrane. Using a novel antibody, we confirm here that the immunocytochemical staining was indeed due to an increase in utrophin at the sarcolemma. The result is observed not only on mdx (an animal model of DMD) myotubes in culture but also in mdx mice treated with L-arginine. In addition, we show here the utrophin increase in muscle extracts of mdx mice treated with L-arginine, after electrophoretic separation and western-blotting using this novel antibody, and thus extending the electrophoretic results previously obtained on myotube cultures to muscles of treated mice. PMID- 11019370 TI - [5'-DFUR chemoprophylaxis in superficial bladder cancer. Kanagawa Urology 5'-DFUR Study Group]. AB - 5'-Deoxy-5-fluorouridine (5'-DFUR), an oral fluorinated pyrimidine carbamate, is widely used in patients with gastrointestinal and breast cancers because of its effectiveness. However, in bladder cancer, response rates have only been reported in Phase II clinical trials. Therefore, we conducted a prospective randomized trial to investigate chemoprophylactic effect of 5'-DFUR against recurrence of superficial bladder cancer after transurethral bladder tumor resection (TUR-Bt). The subjects were grouped as follows: 1) 5'-DFUR group (n = 31), received 600 mg/day of 5'-DFUR starting 2-3 weeks after TUR-Bt for 2 years; and 2) control group (n = 31) received no 5'-DFUR. Although there was no significant difference between groups, the cumulative recurrence rates was more favorable in the 5'-DFUR group (p = 0.256) than in the controls. Results according to cancer factors showed that, in patients with G2 based on grading, those in the 5'-DFUR group tended to have a lower recurrence rate than the control group (p = 0.070). There was a 40% incidence of adverse drug reactions (12/30 patients), primarily slight gastrointestinal symptoms which disappeared or improved with drug discontinuation. The results of the present study suggest that 5'-DFUR might be the choice of treatment to prevent recurrence of superficial bladder cancer. PMID- 11019371 TI - Inhibition of disease flare with diethylstilbestrol diphosphate and chlormadinone acetate administration for two weeks prior to slow-releasing leuprolide acetate in prostatic cancer patients. AB - To determine whether administration of estrogen or gestagen prior to luteinizing hormone-releasing hormone (LH-RH) agonist prevents disease flare in prostate cancer patients, we pretreated the patients with either diethylstilbestrol diphosphate (DES-P) 300 mg daily (N = 17) or chlormadinone acetate (CMA) 100 mg daily (N = 16) or none (N = 16) for two weeks before the initial injection of leuprolide acetate (L). Blood samples for prostatic specific antigen (PSA), testosterone (T), and luteinizing hormone were collected before CMA and DES-P administration, before and at 2, 7, 14, 28, 56, and 84 days after the first administration of leuprolide. The treatment with DES-P and CMA prior to LH-RH agonist induced an early decline of PSA. The mean PSA level showed no significant secondary rise in those patients with pretreatment after L administration. In the patients pretreated with DES-P or CMA, the mean serum T level never exceeded the pretreatment baseline after L administration. On the other hand, in the patients without DES-P or CMA, both serum T and PSA levels increased after the first administration of L. These results clearly demonstrate that pretreatment with DES P 300 mg daily or CMA 100 mg daily for 2 weeks is quite effective to prevent disease flare after the first administration of L in patients with prostatic cancer. PMID- 11019372 TI - [National trend of management of benign prostatic hyperplasia in Japan during 1990s: analysis of national health statistics]. AB - To review the contemporary management of benign prostatic hyperplasia (BPH) in Japan during 1990s, we analyzed several nationwide health statistics by the Ministry of Health and Welfare of Japan. The cross-sectional surveys revealed that the estimated total number of patients receiving treatment increased from 202,000 in 1987, to 335,000 and 590,000, respectively, in 1995 and 1998. Approximately 73-80% of patients were men aged 65 years or over and 94-98% 55 years or older. Urologists treated two thirds of the BPH patients. The incidence of prostatectomies remained relatively stable at 50,000/year (3.0-3.8 prostatectomies/1,000 men aged 55 or over). The average hospital stay in 1996 was 24.7 days. The total cost of BPH therapy nearly doubled between 1988 and 1998. The ratio of outpatient to inpatient costs ranged from 1.5 to 2.2 and 60% of the outpatient cost was spent for medical therapy. The total value of the market for medical therapy increased from 30-40 billion yen in 1989 to more than 80 billion yen in 1998. The application of alpha-blockers increased from 243,000 men (70% of all patients) in 1995 to 452,000 (77%) in 1998, whereas the number of patients taking antiandrogens, plant extracts and antispasmodic agents/Ca antagonists (for pollakisuria), respectively, remained relatively stable at 60,000-70,000, 180,000 and 300,000. Because Japan is a rapidly aging society and men aged 55 or older are expected to increase from 15 million in 1995 to 21 million in 2010, cost effective treatment guidelines for the Japanese BPH patients are needed. PMID- 11019373 TI - [Renal cell carcinoma in a patient with malignant lymphoma: a case report]. AB - A 65-year-old woman was admitted for the treatment of malignant lymphoma. Computed tomography revealed a right renal tumor. After 3 cycles of CHOP (cyclophosphamide, adriamycin, vincristine, prednisone) chemotherapy, we performed right radical nephrectomy. The histopathological diagnosis was renal cell carcinoma. After nephrectomy she was treated with 3 cycles of CHOP chemotherapy and radiation therapy. She received no adjuvant therapy for renal cell carcinoma and had no recurrence after 8 months from the nephrectomy. PMID- 11019374 TI - [Chromophobe cell renal carcinoma: a case report]. AB - We report a case of chromophobe cell renal carcinoma. A 45-year-old woman with a chief complaint of macrohematuria and left backache was introduced to our hospital in October 1998. Abdominal computed tomographic scan (CT) revealed a solitary, solid mass (16 x 14 x 12 cm) at the upper pole of the left kidney. Angiography showed a hypervascular character but irregular neovasculizations were found. We performed a radical nephrectomy with a preoperative diagnosis of malignant renal tumor. The cut surface appearance of the tumor was homogeneous, grey-beige and solid. Light microscopic findings revealed voluminous cells with light, but not clear cytoplasm displaying a fine reticular pattern. Histochemical staining with Hale's colloidal iron stain demonstrated a distinctly positive cytoplasmic reaction. Since the ultrastructural study revealed numerous intracytoplasmic microvesicles, we finally reached the diagnosis of chromophobe cell renal carcinoma. We report our case here with reference to the relevant literature. PMID- 11019375 TI - [A case of squamous cell carcinoma arising in the urinary bladder diverticulum]. AB - A 49-year-old man was admitted to our hospital with the chief compliant of difficulty of urination, lower abdominal pain and macrohematuria. On cystoscopy, a non-papillary tumor was seen in the diverticulum on the right side of the urinary bladder. Having made the diagnosis of urinary bladder cancer, we performed total cystectomy, retroperitoneal node dissection and construction of an ileal neobladder. Histologically, it was squamous cell carcinoma, pT3a, pN0. Seven months after the operation, the patient died of recurrent disease. PMID- 11019376 TI - [Rhabdomyolysis complicated with a fistula between ileal neobladder and intestine: a case report]. AB - A 79-year-old male with an ileal neobladder was hospitalized with the chief complaints of fever, dark colored urine and systemic muscle pain. The clinical diagnosis was rhabdomyolysis complicated with hypokalemia, which was caused by chronic diarrhea due to a fistula between ileal neobladder and intestine. The patient was treated successfully with efficient drip infusion. PMID- 11019377 TI - [Torsion of the spermatic cord in undescended testis: report of two cases]. AB - We report two cases of torsion of the spermatic cord in undescended testis. Case 1: A 23-year-old man was admitted to our hospital with the complaints of fever and left inguinal pain. He had cerebral palsy in his past history. Tentative diagnosis of acute epididymitis of left undescended testis was made, and antibacterial drugs were given. Response was poor, and torsion of the spermatic cord was suspected strongly. Torsion of the spermatic cord in undescended testis and severe testicular infarction were seen in surgery after 13-day conservative treatment. Orchiectomy was performed. Case 2: A 6-year-old boy was admitted to our hospital with the chief complaints of left inguinal mass and pain. He had felt abdominal pain for 3 days. Scrotal contents were impalpable and the diagnosis of torsion of the spermatic cord was made. Orchiectomy was performed due to severe testicular infarction. PMID- 11019378 TI - [A case of tuberculous epididymitis which was difficult to distinguish from a testicular tumor]. AB - A 70-year-old man complaining of painless right scrotal swelling was referred to our hospital. He had a past history of left nephrectomy for renal tuberculosis at the age of 28. Power Doppler ultrasonography revealed a hypoechoic and hypovascular tumor with septa in the scrotum. We suspected a right testicular tumor and therefore, performed a right high inguinal orchiectomy. On macroscopic findings, the fluid of the tumor was yellowish and mucinous and the ipsilateral testis was remarkably atrophic. The pathological diagnosis was tuberculous epididymitis with central necrosis. In recent years, tuberculous epididymitis is rare, and this case was considered to be the first report of power Doppler ultrasonographic findings in tuberculous epididymitis. PMID- 11019379 TI - [The current status of assisted reproductive technology]. AB - It has been over two decades since the birth of the first child conceived by in vitro fertilization (IVF). During the intervening years, technology has evolved; however, IVF has not solved the problems concerning sperm. The first successful pregnancies after the intracytoplasmic injection of a single spermatozoon into an oocyte (ICSI) were rapidly followed by the widespread use of this novel technique for the treatment of male factor infertility. Injection of motile (living) spermatozoa into the oocyte is the most important factor in obtaining good results and other sperm parameters, anti-sperm antibodies and sperm origin, i.e., ejaculated, epididymal and testicular, do not have a strong influence on the outcome of ICSI. ICSI has revolutionized the treatment of male infertility and the application of ICSI is rapidly expanding around the world. However, this technique avoids the natural process of sperm selection and fertilization. Therefore, it is of utmost importance to monitor carefully the development, throughout childhood and into adulthood, of individuals born as a result of ICSI in order to assess its safety. ICSI should only be used for specific indications until its safety has been established. In this paper, the current status of two new approaches, ICSI with spermatid and donor oocyte cytoplasm transfusion, is also reviewed. PMID- 11019380 TI - [Clinical experience with assisted reproductive technology along the development of micro fertilization]. AB - In 1993 we had the first success of micro fertilization in our clinic. The method by partial opening of oocyte zona pellucida initially used achieved a fertilization rate per egg of only 26.7%. Although transfer of sperm under the zona raised the fertilization rate to 39.2%, the new method involved problems such as polyspermy, which represented approximately 30% of the total fertilization rate. In addition, use of this method was limited to patients meeting several conditions. In 1994, we had a successful case of pregnancy after intracytoplasmic sperm injection (ICSI). The ICSI method resulted in a fertilization rate of about 70%, and has solved the problem of polyspermy. Fertilization of eggs has become possible using immotile sperm, and the technique of microsurgical epididymal sperm aspiration (MESA) and testicular sperm extraction (TESE) has been established. Clinical data have confirmed that a fertilization rate by frozen sperms is not lower than that by fresh sperms. Sperm can be stored frozen after MESA or TESE and therefore are surely available when needed, allowing to avoid spoiling retrieved eggs. No difference in fertilization rate was found between conditions needing micro fertilization or sperm recovery methods. No difference in pregnancy rate was shown between treatment cycles. Recent reports show improved pregnancy rates using techniques such as transferring a frozen embryo after endometrical adjustment. Further development in assisted reproductive technology is anticipated. PMID- 11019381 TI - [Epididymal sperm aspiration for obstructive azoospermia]. AB - Epididymal sperm aspiration technique combined with assisted reproductive technology (ART) including intracytoplasmic sperm injection (ICSI) has provided new frontiers for the treatment of unreconstructable obstructive azoospermia, including congenital bilateral absence of the vas deferens and failed surgical intervention. Epididymal sperm is obtained by several procedures, including microsurgical epididymal sperm aspiration (MESA), mini-MESA (Modified MESA), macroscopic epididymal sperm aspiration (MaESA) and percutaneous epididymal sperm aspiration (PESA). Since 1991 in our department, epididymal sperm aspiration combined with ART was performed by MESA (26 cases, 41 times) and PESA (3 cases, 4 times). Motile sperm retrieval using MESA and PESA was obtained in 34 out of 36 times (94.4%) and 2 out of 4 times (50%), respectively. MESA-ICSI resulted in a 47.3% 2PN (metaphase II) fertilization rate per eggs and a 28.8% pregnancy rate per transfer. With advancement of ICSI technique in which frozen sperm can be used, elective sperm retrieval can be planned at our convenience. PESA is a convenient, inexpensive and effective outpatient clinic procedure for retrieving sperm assisted with ICSI. In summary, infertile couples need to be given realistic options regarding treatment outcome. The fertility potential and age of the female partner need to be considered when addressing male treatment options. PMID- 11019382 TI - [Retrieval and cryopreservation of testicular sperm]. AB - We reviewed 44 cases of non-obstructive azoospermia treated by testicular sperm extraction and intracytoplasmic sperm injection (ICSI) from July, 1997 to September 1999 at our institutes. Testicular sperm were retrieved from 32 patients (72.7%). ICSI was performed on 29 patients and the partners of 15 patients (46.9%) became pregnant. Out of 10 patients with histology of Sertoli cell-only, we could retrieve sperm from 3 patients (30%). Testicular volume, Johnsen's score count (JSC), and FSH were significant parameters to predict the recovery of testicular sperm from the patients, but if only the patients with JSC less than 8, are analyzed, none of them was significant parameter. Chromosomal abnormality was not a significant parameter. The partner's age, motility of recovered sperm and testicular volume correlated with the fertilization rate. Chromosomal abnormality or the usage of cryopreserved sperm was not a significant parameter to predict fertilization. PMID- 11019383 TI - [Effects of the gene transfer into sperm mediated by liposomes on sperm motility and fertilization in vitro]. AB - To clarify the problems of gene transfer in the fertility clinic, we investigated the safety and effectiveness of the gene transfer of mouse sperm using the lipofection method. We used the pCAGGS-lacZ plasmid connected with cytomegarovirus enhancer/chicken beta-actin promoter as an adventitious gene. After incubation of the DNA-liposome complex and mouse sperm of epididymis in the medium, we investigated the sperm motility after gene transfer, in vitro fertilization rate of the gene transferred sperm, and gene expression of the fertilized ovum and fetus. Sperm motility was lowered significantly with the increase in DNA density. The fertilization rate for in vitro fertilization was lowered significantly with the increase in DNA density. Gene expression was observed in some of the fertilized ova. In the fetus 12.5 days after embryo transfer, gene transfer was confirmed by the PCR method, but we could not find gene expression by X-gal staining. We proved that the sperm with a gene transferred by the lipofection method could fertilize the ovum and transfer adventitious gene to ova. We speculate that sperm motility and fertility rate were lowered because the introduction of DNA and the DNA-liposome complex destroyed parts of the cell. Development of a sperm fractionation method for DNA transfer and establishment of a method for effective and safe DNA transfer are awaited. PMID- 11019384 TI - [Antipseudomonal activity of carbapenem antibiotics]. AB - To date, three carbapenem antibiotics have been introduced for clinical use, and they can be structurally classified into two types. One is a natural type that has the naturally-occurring carbapenem skeleton and a strongly basic (cationic) moiety in the C-2 side chain, like imipenem or panipenem. The other is a new generation carbapenem, meropenem, which has the 1 beta-methyl carbapenem skeleton and a less basic group in the C-2 side chain. It was reported that there were some significant differences among these two types of carbapenems concerning the antimicrobial profile, especially the antipseudomonal activity. Since Pseudomonas aeruginosa was one of the target pathogens of carbapenem antibiotics, these facts prompted us to overview the different mode of action among imipenem, panipenem and meropenem and clarify the structure-activity relationships of carbapenems with regard to the antipseudomonal activities. In this article, we discuss that both the chemical structure and the physicochemical properties of carbapenems greatly influence a variety of antipsedomonal actions including MIC, affinity for PBPs, outer membrane permeability, interaction with various beta-lactamases and multidrug efflux systems etc., and that the cationic center in the C-2 side chain plays an important role in antipseudomonal activities. This review will be helpful in developing new types of antipseudomonal carbapenems and/or new clinical applications of carbapenem antibiotics for treating pseudomonal infection. PMID- 11019385 TI - [Antimicrobial susceptibility and serotypes of Streptococcus pneumoniae isolated from the patients with respiratory tract infections in Hokusetsu Area of Osaka]. AB - One hundred and thirteen strains of Streptococcus pneumoniae (S. pneumoniae) were isolated from the clinical specimens of patients with respiratory tract infections between January and December 1998 in three hospitals in Hokusetsu area of Osaka. We investigated susceptibility of 113 strains of S. pneumoniae to benzylpenicillin (PCG) and other antimicrobial agents and their serotypes. 1) Of the 113 strains of S. pneumoniae isolated, 25.7% were susceptible (PSSP), 51.3% were intermediate (PISP) and 23% were resistant to benzylpenicillin (PRSP). 2) The MICs of cefaclor, cefditoren, cefpodoxime, cefdinir, erythromycin, clindamycin and minocycline were elevated, but the MIC values of cefditoren ranged from < or = 0.03 to 1.0 microgram/ml. The susceptibility of 113 strains to cefditoren was comparatively high. 3) The MIC values of imipenem, meropenem and vancomycin for 81 strains of PISP and PRSP ranged from < or = 0.015 to 1.0 microgram/ml, from < or = 0.015 to 2.0 micrograms/ml and from 0.13 to 0.5 microgram/ml, respectively. The susceptibility of these strains to three antimicrobial agents was superior to that to the other antimicrobial agents examined. 4) Of the 60 strains examined, 19, 6, and 23 serotypes were 30, 25 and 18.3%, respectively. The three serotypes were observed in PISP and PRSP with a high frequency. 5) Isolates of S. pneumoniae were 37.2% for children under 2 years of age and 30.9% for children from 2 to 6 years of age. Most of the strains isolated from these children were resistant. PMID- 11019386 TI - [Antibacterial activity of fosfomycin against the causative bacteria isolated from bacterial enteritis]. AB - The in vitro antibacterial activities of fosfomycin (FOM) and 3 fluoroquinolones against Salmonella spp., pathogenic Escherichia coli, Campylobacter spp. and Shigella spp. were investigated. The activity upon the environmental condition in the inflammation was compared with standard condition in vitro. On standard condition, the MIC90 of tosfloxacin (TFLX), norfloxacin (NFLX) and levofloxacin (LVFX) against E. coli (77 strains), Shigella spp. (50) and Salmonella spp. (41) were < or = 0.025-0.10, 0.10, and 0.05 microgram/ml, respectively. The MIC90 of FOM against those organisms was 0.39-1.56 micrograms/ml. The MIC90 of TFLX, NFLX, LVFX against Campylobacter spp. were 6.25, 100 and 3.13 micrograms/ml, respectively. The MIC90 of FOM was 50 micrograms/ml. The activity of FOM was unaffected by pH and in anaerobic condition. On the other hand, the activity of NFLX was decreased in low pH and in anaerobic condition. In the presence of horse blood and addition of Na+, the activities of both agents were unaffected. These results suggested that FOM is equally active with or superior to fluoroquinolone in the intestinal infection treatment. PMID- 11019387 TI - Effect of mite antigens on antigen presenting cells/macrophages in mice. AB - The effect of mite antigens on murine lymphocytes and macrophages was studied in vitro. Antigens prepared from Dermatophagoides farinae bodies (Dfb) or recombinant Mag3, glutathione-S transferase (GST)-fused mite antigen, stimulated murine spleen cells to proliferate. The responder cells were B cells, because the response was sensitive to anti-Ig antibody and C treatment, but not to anti-Thy 1 antibody and C treatment. The response was not due to lipopolysaccharide contamination, a representative B-cell mitogen, because polymyxin B column-passed Dfb significantly stimulated B cells, and GST protein alone did not stimulate them. Alloantigen presenting activity was increased in mite antigen-treated B cells and spleen adherent cells. Mite antigens stimulated CD80 and the major histocompatibility complex (MHC) class II molecule expression, but suppressed CD86 expression on B cells and spleen adherent cells that were detected by a flow cytofluorometer. Antibodies to the MHC class II molecules, CD80 and CD86 blocked the alloantigen-presenting activity. Furthermore, mite antigens stimulated B cells and spleen adherent cells to produce cytokines. These results suggest that mite antigens have a stimulating activity on antigen-presenting cells/macrophages and modulate immune responses. PMID- 11019388 TI - Evaluation of dietary intake using carbon and nitrogen isotope analysis of human hair of Chinese living in southern part of China. AB - We examined dietary intakes among 69 people of three ethnic groups (Yao Chinese, Zhung Chinese and Han Chinese) in the autonomous country of Bama in China where many elderly people have a long life span in 1998. Dietary intake was evaluated using carbon and nitrogen isotope ratios of human hair (delta 13C, delta 15N) standardized by standard gas. The distributions of delta 13C in Yao Chinese and Han Chinese were scattered from -21 to -17/1000, which were close to that of C4 plants such as corn and millet. The distribution of delta 13C in Zhung Chinese was located from -25 to -22/1000, which was near that of C3 plants such as rice, wheat and potato. These values showed a much wider range when compared with delta 13C in Chinese living in urban areas. On the other hand, delta 15N in each group was distributed in the area ranged from 8 to 11/1000. The result of delta 13C was consistent with data obtained from a nutritional survey, but that of delta 15N was a little higher than expected. The carbon and nitrogen isotope ratios of human hair among three ethnic groups showed characteristic distributions, although the relation between these isotope ratios and longevity is uncertain. PMID- 11019389 TI - Idiopathic postural orthostatic tachycardia syndrome (POTS): emerging concepts for the clinician. PMID- 11019390 TI - [Idiopathic postural orthostatic tachycardia syndrome] AB - Stimulated by the widespread use of head-up tilt testing, transient episodes of neurocardiogenically mediated hypotension and bradycardia have become a well recognized cause of recurrent syncope and near syncope (generally referred to as neurally mediated syncope). On the other hand, a large subgroup of patients was identified, who appeared to have a less severe hypotension and orthostatic intolerance that is characterized by postural tachycardia, exercise intolerance, disabling fatigue, lightheadedness and dizziness. This form of disability has been recognized as postural orthostatic tachycardia syndrome (POTS). While the etiology of POTS is still unclear, a mild form of idiopathic peripheral autonomic neuropathy (partial dysautonomia) or beta-receptor hypersensitivity has been suggested for the pathophysiology of this disorder. A detailed history and physical examination that includes a careful neurologic examination are essential for diagnosis. Head-up tilt testing is often useful as a standardized measure of response to postural change. This review summarizes the history, current knowledge of clinical features, diagnosis and therapeutic strategies. PMID- 11019391 TI - [Development and evaluation of the health management system including medical images]. AB - In the field of occupational health management, there are many areas in which a number of X-ray films that are associated with health checkup data for longitudinal health management of high risk groups exposed to dusts and specific carcinogenetic chemicals can be used. To make comprehensive use of the medical checkup data, we examined and evaluated basic techniques in which medical images such as radiographs can be linked to the medical checkup data in a form of computer data. A three-step procedure system was developed based on the IS&C system which is a common specification of medical images: 1) data convert and barcode printing system, 2) film digitizing and database management system, 3) utilizing system of health checkup data including a medical image system. Experiments were done using the system and it was found that processing efficiency seemed to be less and/or varied depending on process types. It was also found that the extraction of data from existing systems and the storage methods of data onto a magnetooptical disk are special issues to be resolved in our system. PMID- 11019392 TI - [Seven cases of radiation-induced cutaneous squamous cell carcinoma]. AB - We report 7 cases of radiation-induced skin cancer. The diagnosis was based on the history of radiotherapy for benign skin diseases (5 cases) and of occupational exposures to medical doctors (2 cases). All cases were squamous cell carcinomas which arose from chronic radiodermatitis. The estimated latent period of these tumors ranged from 6 to 64 years, with an average of 29.9 years. After surgical treatments of the lesions, no local recurrences were observed in all cases. Benign skin diseases had sometimes been treated with low-energy radiation before the 1960s. Considering the estimated latent period, the peak time point of developing risk of radiation-induced skin cancer by such treatment has been already passed, however, the danger of it should not be ignored in future. In association with multiplicity of radiation usage, occupational exposure of radiation may develop the risk of occurrence of skin cancer in future. Therefore, we should recognize that radiation-induced skin cancer is not in the past. In the cases of chronic skin diseases showing warty keratotic growth, erosion and ulcer, we should include chronic radio-dermatitis in the differential diagnosis. It is necessary to recall all patients about the history of radiotherapy or radiation exposure. Rapid histopathological examination is mandatory because of the suspicion of radiation-induced skin cancer. PMID- 11019393 TI - [Molecular mechanisms of multidrug resistance in Mycobacterium tuberculosis]. AB - Control of tuberculosis caused by multidrug-resistant (MDR) Mycobacterium (M.) tuberculosis has become one of the major problems throughout the world. Understanding of the molecular mechanisms of resistance may help in the development of novel methods for the rapid and precise detection of drug resistant M. tuberculosis. Eight agents have been recommended to treat tuberculosis. Isoniazid (INH), rifampicin (RFP), pyrazinamide (PZA), streptomycin(SM), and ethambutol (EB) are used as the first line agents, and the others are the second line agents. MDR M. tuberculosis strains are resistant both to INH and RFP which have the most effective bactericidal activity to M. tuberculosis. Nearly 95% of RFP resistant strains possess a mutation on the rpoB gene encoding a DNA-dependent RNA polymerase. INH particularly shows an inhibition of the cell wall synthesis of M. tuberculosis and approximately 90% of INH resistant strains have a mutation on the inhA, katG, and ahpG gene encoding enzymes related to a mycolic acid synthesis of cell wall. PZA resistant strains have a mutation on the pncA gene encoding a pyrazinamidase which degradates pyrazinamide to a bactericidal substance, pyrazinoic acid. SM resistant strains have a mutation on the rrs and rpsL gene encoding a 16S rRNA and a S12 ribosomal subunit protein, respectively. EB resistant strains have a mutation on the embB gene encoding a arabinosyl transferase which catalyzes cell wall synthesis. Resistant mechanisms of second-line agents have also been identified. Recently, rapid detection methods for RFP and INH resistant mutations have been developed on the basis of these studies. PMID- 11019394 TI - [A report on symposium, "protection for personal information in epidemiological research and cancer registry"]. AB - A symposium on protection of personal information in epidemiological research and the compilation of a cancer registry was held at the National Cancer Center, Japan, on March 16, 2000. Participants discussed the use of personal health information in epidemiological research activities such as the compiling of a cancer registry to protect personal information. Legislation in Japan is pending in the near future. A total of thirteen domestic and foreign speakers and commentators shared opinions from various viewpoints and discussed the issue. It was proposed that protection regarding personal information should be legislated as soon as possible for the development of cancer registry; patients could be exempted from informed consent for utilization of personal health information, in particular, in public health activities such as the complication of a cancer registry and epidemiological research with the approval of an ethics review board; ethical guidelines including these steps should be established in Japan. An appropriate system of protection of personal information should be established through constructive discussion at frequent meetings and symposia which are open to the citizenry. PMID- 11019395 TI - [Report from the 7th UOEH meeting of gastrointestinal image diagnosis]. PMID- 11019396 TI - [Report from the 9th UOEH meeting of gastrointestinal image diagnosis]. PMID- 11019397 TI - Lymph and lymphangiogenesis: functional aspects. PMID- 11019398 TI - An avian model for studies of embryonic lymphangiogenesis. AB - Embryonic development of lymphatics (lymphangiogenesis) in recent years has rarely been studied experimentally. Using an avian model, we showed that both intra- and extra-embryonic blood vessels of chick and quail embryos are accompanied by lymphatics. The lymphatics of the chorioallantoic membrane (CAM) are drained by lymphatic trunks of the umbilicus and are connected to the posterior lymph hearts. Intra-embryonic lymphatics are drained via paired thoracic ducts into the jugulo-subclavian junction. The lymphatic endothelial cells are characterized by the expression of Vascular Endothelial Growth Factor Receptors (VEGFR) -2 and -3. Application of VEGF-C, the ligand of these two receptors, on the differentiated CAM, induces proliferation of lymphatic endothelial cells and formation of huge lymphatic sinuses. These lymphatics derive from pre-existing lymphatic endothelial cells, whereas, in early embryos lymphangioblasts are present in the mesenchyme. This phenomenon can be demonstrated by interspecific grafting experiments between chick and quail embryos. Together with the early lymph sacs, the lymphangioblasts form the embryonic lymphatic system. Our studies demonstrate the importance of lymphangioblasts and lymphangiogenic growth factors in embryonic lymphangiogenesis. PMID- 11019399 TI - Ignorance in infectious diseases: the case of AIDS, Kaposi sarcoma, and lymphology. AB - From the perspective of The University of Arizona's innovative Curriculum on Medical (and Other) Ignorance focusing on "what we know we don't know, don't know we don't know, and think we know but don't," the shifting terrain of information knowledge-ignorance of AIDS (a disorder involving, to various incompletely understood degrees, the four components of the lymphatic system-lymph, lymphatics, lymphocytes, and lymph nodes) and Kaposi sarcoma (a lymphedemogenic lesion thought to arise from trans-differentiated lymphatic endothelium) is surveyed by pinpointing some key unanswered questions that have been raised over the course of the epidemic and pointedly in past International Congresses of Lymphology. These questions are placed in the context of general ignorance about infectious diseases and the relationship of "germ" to "terrain" through the "blood-tissue-lymph loop." A framework is suggested for an "ignorance agenda" encompassing basic biology, clinical management, and societal issues. PMID- 11019400 TI - Non-invasive assessment of the lymphedematous limb. AB - Accurate assessment of the swollen limb is crucial to effective management, and usually consists of measurement of volume and assessment of skin condition. Here, we review the different methods available to measure volume, and their accuracy, together with other non-invasive methods available to assess the characteristics of the swelling. These include the measurement of fluid mobility by recording deformation of tissue by a mass (tonometry) and the step compression method; the measurement of truncal swelling by skinfold calipers; imaging techniques (magnetic resonance imaging, computed tomography, ultrasound) which provide information on size and other characteristics of the different tissue compartments; and measurement of impedance (amount of extracellular water and total water content). The varying quality of swelling, as well as its extent and distribution, indicates the need for objective methods of assessment other than simple limb volume measurement. Such detailed information should improve the understanding of peripheral lymphedema. PMID- 11019401 TI - Bioelectrical impedance analysis revisited. PMID- 11019402 TI - Kudos for chest radiography article. PMID- 11019403 TI - Schizophrenia DR fills gap. PMID- 11019404 TI - Ultrasound assessment of uterine disorders. AB - Around the time of menopause many women develop disorders of the uterus that may cause abnormal bleeding and other troublesome symptoms. Some of these disorders, such as uterine polyps, are typically benign and easily correctable. Others, such as endometrial cancer, are potentially more serious. This article discusses the etiology, diagnosis and treatment of some common uterine disorders, with special emphasis on their sonographic appearance. PMID- 11019405 TI - The mammographer's role in addressing special needs. AB - Many mammography patients have special needs related to their age, sex, surgical status or a physical or mental condition. This article describes some of the accommodations mammographers can make to ensure excellent imaging and quality care for their patients with special needs. PMID- 11019406 TI - The role of the physician extender in radiology. AB - This article examines the role of the radiology practitioner assistant (RPA), a physician extender first defined in the 1970s. The purpose of the research was to determine RPAs' accuracy in recognizing abnormal image patterns and their ability to independently perform gastrointestinal fluoroscopic procedures. The results support previous research indicating that technologists with additional education, training and radiologist supervision can detect abnormal image patterns and perform fluoroscopy successfully. PMID- 11019408 TI - 2000 John B. Cahoon, Jr. Memorial Lecture. Hello in there. PMID- 11019407 TI - Boerhaave syndrome. AB - Boerhaave syndrome is a serious and rapidly fatal spontaneous esophageal rupture that mimics myocardial infarction or upper gastrointestinal conditions. Initially misdiagnosed about 50% of the time, Boerhaave syndrome has a mortality rate of more than 90% without prompt diagnosis and treatment. This article presents a case of nearly fatal Boerhaave syndrome in an otherwise healthy young adult and discusses the historical, statistical, pathophysiological, diagnostic and therapeutic aspects of the disease. PMID- 11019409 TI - To maintain and disseminate. PMID- 11019410 TI - Chalk it up to experience. PMID- 11019411 TI - Avoiding patient falls in radiology. PMID- 11019412 TI - Quality assurance in CT. PMID- 11019413 TI - Managing change in radiology. PMID- 11019415 TI - Computed tomography. PMID- 11019414 TI - Improving radiography's image. PMID- 11019416 TI - [Current approaches to the treatment of chronic viral liver diseases]. PMID- 11019417 TI - [Current approach to the treatment of acute bronchial asthma]. AB - AIM: To evaluate the results of nebulised berodual solution in comparison with intravenous euphylline in the treatment of acute bronchial asthma (BA). MATERIAL AND METHODS: The trial entered 73 patients with an acute episode of BA: 22 treated with intravenous euphylline and 51 treated with nebulised berodual solution. Spirometry was conducted and peak expiratory flow rate, heart rate (HR), blood pressure were measured before and after the treatment. RESULTS: The attack was arrested in all the patients who had inhaled nebulised berodual solution. No side effects were registered. Intravenous therapy of euphylline produced only partial bronchodilation in most of the patients and increased HR by 23.4%. CONCLUSION: Berodual nebulised solution is clinically superior to intravenous euphylline in acute BA. PMID- 11019418 TI - [Clinical and cost effectiveness of nebulizer therapy in emergency care for acute bronchial asthma]. AB - AIM: To evaluate clinical and cost efficacy of urgent therapeutic measures for patients with acute bronchial asthma (BA) in a large industrial city. MATERIAL AND METHOD: 380 patients with moderate and severe BA in emergency state were observed. 222 of them got berodual solution provided by pneumatic nebuliser, moistened oxygen and corticosteroids. 158 patients received standard emergency care. Bronchopulmonary and cardiovascular systems were studied. RESULTS: The proposed therapy proved 1.6 and 2.6 times more effective in moderate and severe BA, respectively, than standard therapy. It made possible to reduce the treatment cost by 30%. An algorithm for emergency care for acute BA is designed. CONCLUSION: Differential emergency care is recommended for patients with acute bronchial asthma. PMID- 11019419 TI - [Analysis of bronchial hypersensitivity in patients with bronchial asthma]. AB - AIM: To analyse correlation between bronchial hypersensitivity in bronchial asthma (BA) and environmental factors; to propose methods of relevant correction. MATERIAL AND METHODS: The trial included 97 BA patients (42 males, 55 females) aged 17-79 years. Bronchial sensitivity was studied by means of dose-dependent bronchial resistance curve (Rrs) plotted by the unit Masterlab (Germany) in the course of long-term inhalation of a gradually rising dose of methacholine. A regression analysis was made of the dependence of fluticasone bipropionate effectiveness on clinical symptoms of the disease. RESULTS: Bronchial hypersensitivity depends on initial clinical symptoms of BA, in a less degree on bronchial permeability, severity of inflammation of bronchial mucosa. CONCLUSION: Local corticosteroid fluticasone propionate is a drug of choice in the treatment of mild and moderate BA. PMID- 11019420 TI - [Clinical aspects of trimetazidine (preductal) administration as an antianginal drug]. AB - AIM: To study antianginal effectiveness of trimetazidine (TMS) in monotherapy and combined antianginal therapy; to design approaches to prediction of its effect. MATERIAL AND METHODS: An open cross-over randomised trial enrolled 29 anginal patients. They were given a single TMS dose (60 mg) and its effect was compared to that of placebo, propranolol, verapamil, isosorbide dinitrate, nifedipine). The effect was estimated by an increment of bicycle exercise (BE) duration necessary to depress ST segment by 1 mm. 10 patients retained signs of myocardial ischemia at BE though they were treated with specially selected antianginal drugs. In them, we assessed additional benefit of adjuvant TMS. The drugs were tried under acute test and one-month treatment. RESULTS: In acute test a significant antianginal effects of TMS were registered against the placebo. Differences in efficiency of the antianginal drugs were insignificant. The addition of TMS to the selected drugs gave rise to the trend to prolongation of time to ST segment depression by 1 mm. We found a strong correlation between TMS efficiency in acute pharmacological test and in long-term administration. This enables using BE for prediction of long-term effect of the drug. CONCLUSION: TMS has a significant antianginal effect. It is especially beneficial in cases of insufficient effectiveness of conventional antianginal drugs or when their use is problematic because of marked disturbances of hemodynamics. PMID- 11019421 TI - [Effect of L-arginine on platelet aggregation, endothelial function adn exercise tolerance in patients with stable angina pectoris]. AB - AIM: Examination of the action of donor NO (L-arginine) on platelet aggregation, endothelial function and exercise tolerance in patients with stable angina of effort (SAE). MATERIAL AND METHODS: 42 patients with SAE (functional class I-II) and 10 healthy volunteers (control group) were assigned to two groups. 22 patients of group 1 were randomized to cross-over. They received cardiket (60 mg/day for 10 days or cardiket (60 mg/day) in combination with L-arginine (15 g/day for 10 days). 20 SAE patients of group 2 and control group received L arginine (15 g/day for 10 days). In each group blood lipids were examined, and bicycle exercise test (BET) was performed. In addition, platelet aggregation and endothelial function were studied in group 2 and control group before and after the course of L-arginine. RESULTS: Compared to control group, endothelial function significantly improved in group 2 (from 5.0 +/- 2.9 to 7.8 +/- 4.1% vs 7.1 +/- 1.9 to 6.6 +/- 4.8%) (M +/- SD). BET duration increased in all the patients. After ADP addition in concentrations 1.5, 2.0, and 5.0 micromol/l platelet aggregation declined in 17 patients except 3 in whom the aggregation remained unchanged. CONCLUSION: Positive effect of L-arginine on endothelial function, exercise tolerance and platelet aggregation was observed in patients with stable angina of effort (functional class I-II). Therefore, arginine can be recommended as an adjuvant in the treatment of patients with ischemic heart disease. PMID- 11019422 TI - [Effect of probucol on cardiac electrophysiology in anginal patients with hyperlipidemia and diabetes mellitus type II]. AB - AIM: To investigate effects of a hypolipidemic drug with antioxidant action probucol on electrophysiological parameters of the heart in patients with ischemic heart disease (IHD), stable angina (SA), hyperlipoproteinemia (HLP) and diabetes mellitus (DM) type II. MATERIAL AND METHODS: The trial entered 48 IHD patients (11 males and 37 females) aged 47-73 years with SA functional class II and III (39.4 and 60.6%, respectively), secondary HLP (mean group total cholesterol 6.5 +/- 0.17 mmol/l), DM type II (mean fast glucose 7.7 +/- 1.8 mmol/l) and obesity (mean body mass index 29.7 +/- 2.2 kg/m2). Transesophageal pacing of the left ventricle was conducted in all the patients before probucol treatment and 18 hours, 1 month and 3 months after it. RESULTS: Probucol (a single dose 500 mg) significantly reduced the time of recovery of sinus node function. Three months of probucol administration in a dose 1000 mg/day enhanced pacemaker activity of the sinus node. CONCLUSION: In addition to a beneficial effect on lipid metabolism (a decrease of total cholesterol by 13.4%, LDLP cholesterol by 15.2%, triglycerides by 15.8%), probucol enhanced pacemaker activity of the sinus node. This makes it perspective in patients with sinus node dysfunction. Caution is necessary in prescribing probucol to patients with supraventricular tachycardia caused by re-entry mechanism. PMID- 11019423 TI - [Effect of manual therapy on clinico-pathophysiological status and central hemodynamics in patients with ischemic heart disease]. AB - AIM: To elucidate effects of manual therapy on clinical status, central hemodynamics and parameters of Holter monitoring in ischemic heart disease (IHD) patients with angina of effort. MATERIAL AND METHODS: The examination of 70 IHD patients (functional class II-III) included Kubicek tetrapolar chest rheography in modification of Yu. G. Pushkar, bicycle exercise, 24-h Holter ECG monitoring. The condition of the vertebromotor segments was evaluated at manual tests. RESULTS: The addition of manual therapy to combined treatment of IHD patients improves sympathetic innervation in the cervicothoracic segments, depresses myofascial hypertonicity, diminishes sympathicotonic effects on the heart, enhances myocardial, coronary and aerobic reserves. CONCLUSION: The results of the study allow recommending adjuvant manual therapy in combined treatment of IHD. PMID- 11019425 TI - [Platelet aggregation in patients with hypertrophic cardiomyopathy]. AB - AIM: To examine correlation between platelet aggregation and the degree of left ventricular hypertrophy (LVH) in patients with hypertrophic cardiomyopathy (HC). MATERIAL AND METHODS: Routine clinical examination, ECG, echo-CG, measurement of platelet aggregation, routine biochemical blood tests were made in 45 HC males and 15 healthy controls. RESULTS: HC patients were characterized with high induced and spontaneous platelet aggregation which increased with left ventricular myocardial mass. In LVH, platelets were especially sensitive to the aggregation inductor U-46619, thromboxane A2 analogue. CONCLUSION: It is demonstrated that there is a positive correlation between platelet aggregation and the degree of LVH in HC patients. PMID- 11019424 TI - [Diovan effectiveness in combined treatment of severe arterial hypertension]. AB - AIM: To assess effectiveness of diovan, a novel angiotensin-II antagonist, in combined hypotensive treatment of patients with severe arterial hypertension. MATERIAL AND METHODS: 15 patients with severe arterial hypertension received diovan in a dose 80-160 mg/day for 8 weeks in combination with clofelin, beta adrenoblockers, ACE inhibitors, diuretics prescribed before the start of diovan. Diovan effects were studied by 24-h monitoring of arterial pressure, tolerance, changes in blood counts, central hemodynamics. RESULTS: Adjuvant hypotensive treatment with diovan lowered arterial pressure to satisfactory levels in 80% of patients. 13.3% did not respond to diovan. 6.7% of patients were withdrawn because of allergy. Only 2 patients of those who completed the diovan course developed side effects in first two weeks of the treatment--they had head ache. CONCLUSION: Diovan is effective in combination with clofelin, ACE inhibitors, diuretics, beta-adrenoblockers. PMID- 11019426 TI - [Characteristics of drug resistance of tumor plasmocytes in vitro in patients with multiple myeloma differently responsive to chemotherapy]. AB - AIM: To determine sensitivity of tumor plasmocytes in vitro to cytostatic drugs (prednisolone, alkeran belustin, vincristine, rubomycin, doxorubicin, cytarabin, methotrexate, cysplatin, etoposide). MATERIAL AND METHODS: The sensitivity was measured with DISC method in 12 patients with multiple myeloma (MM) in two groups: resistant and responsive to induction polychemotherapy (PCT). RESULTS: The groups appeared significantly different by lowering of pathological paraprotein concentration (PIg): by 7.4 +/- 2.5% and 32.5 +/- 3.7%, respectively (p < 0.05). The resistance to the drugs was higher in the resistant patients than in the responders (0.7 +/- 0.28 versus 0.4 +/- 0.02, p < 0.05). PCT schemes of resistant patients contained 65.0 +/- 2.3% of ineffective drugs. In the responders the percentage was 35.7 +/- 5.3% (p < 0.05). CONCLUSION: The relationship exists between resistance of tumor plasmocytes to drugs in vitro and clinical findings. PMID- 11019427 TI - [Fludarabine treatment of chronic lymphoid leukemia]. AB - AIM: To try effectiveness of fludarabine monotherapy in verified chronic lymphoid leukemia (CLL) in pretreated patients. MATERIAL AND METHODS: The treatment effectiveness according to the international criteria of complete and partial remission was assessed in 37 patients with progressive, splenic, tumor and bone marrow forms of CLL given 5-day courses of fludarabine in a single daily dose 25 mg. RESULTS: In progressive CLL, the remission was achieved in 45% of the patients (10% complete and 35% partial), in splenic form--62.5% (12.5% complete, 50% partial), in tumor form--50% (100% partial). In bone marrow form no remissions were obtained. CONCLUSION: Fludarabine is effective in therapy of pretreated CLL patients. The treatment should be adjusted to CLL form. PMID- 11019428 TI - [Treatment of local Hodgkin's disease with massive mediastinal involvement]. AB - AIM: To define the scope of combined therapy in patients having local Hodgkin's disease (LHD) with massive mediastinal lesion. MATERIAL AND METHODS: From 1980 to 1997 the treatment was given to 75 LHD patients with massive lesion of the mediastinum (mediastinal-thoracic index, MTI, > or = 0.33). The patients have received combined chemoradiotherapy according to 3 cycle CVPP (cyclophosphamide 600 mg/m2, vinblastine 6 mg/m2, prokarbasin 100 mg/m2, prednisolone 40 mg/m2) with a 2-week interval + radiation to all the lymphatic collectors above the diaphragm with a single focal dose 2 Gy to total focal dose 36-38 Gy + 3 cycle CVPP. According to the size of the mediastinum the patients were divided into 3 groups: group 1--MTI = 0.33, 27 patients; group 2--MTI = 0.33 > 0.45, 32 patients; group 3--MTI > or = 0.45, 16 patients. The response was estimated according to the latest EORTC recommendations. Survival curves were plotted according to the E.L. Kaplan and P. Meier procedure. RESULTS: Complete remission was achieved in 93, 94 and 56% of group 1, 2 and 3 patients, respectively. 6-year recurrence-free survival reached 84, 71 and 32%, while survival free of treatment failure 80, 66 and 27%, respectively. 6-year overall survival and uneventful survival made up 94, 97 and 70%; 72, 66 and 27%, respectively. CONCLUSION: Treatment of LHD patients of group 1 is performed according to the standard scheme (6 cycles of first-line polychemotherapy plus radiotherapy). Patients of group 2 can be treated more intensively, while for patients of group 3 more intensive treatment is a treatment of choice. PMID- 11019429 TI - [Randomized placebo-controlled trial of orlistat for weight loss and prevention of weight regain in obese patients]. AB - BACKGROUND: We undertook a randomised controlled trial to assess the efficacy and tolerance of orlistat, a gastrointestinal lipase inhibitor, in promoting weight loss and preventing weight regain in obese patients over a 2-year period. METHODS: 743 patients (body-mass index 28-47 kg/m2), recruited at 15 European centres, entered a 4-week, single-blind, placebo lead-in period on a slightly hypocaloric diet (600 kcal/day deficit). 688 patients who completed the lead-in were assigned double-blind treatment with orlistat 120 mg (three times a day) or placebo for 1 year in conjunction with the hypocaloric diet. In a second 52-week double-blind period patients were reassigned orlistat or placebo with a weight maintenance (eucaloric) diet. FINDINGS: From the start of lead-in to the end of year 1, the orlistat group lost, on average, more bodyweight than the placebo group (10.2% [10.3 kg] vs 6.1% [6.1 kg]; LSM difference 3.9 kg [p < 0.001] from randomisation to the end of year 1). During year 2, patients who continued with orlistat regained, on average, half as much weight as those patients switched to placebo (p < 0.001). Patients switched from placebo to orlistat lost an additional 0.9 kg during year 2, compared with a mean regain of 2.5 kg in patients who continued on placebo (p < 0.001). Total cholesterol, low-density lipoprotein (LDL) cholesterol, LDL/high-density lipoprotein ratio, and concentrations of glucose and insulin decreased more in the orlistat group than in the placebo group. Gastrointestinal adverse events were more common in the orlistat group. Other adverse symptoms occurred at a similar frequency during both treatments. INTERPRETATION: Orlistat taken with an appropriate diet promotes clinically significant weight loss and reduces weight regain in obese patients over a 2-year period. The use of orlistat beyond 2 years needs careful monitoring with respect to efficacy and adverse events. PMID- 11019430 TI - [Acarbose (glucobai) effects on lipid metabolism in patients with non-insulin dependent diabetes mellitus]. AB - AIM: To investigate the effect of acarbose, intestinal alpha-glucosidase inhibitor, on lipid metabolism in patients with non-insulin-dependent diabetes mellitus (NIDDM). MATERIAL AND METHODS: Glucobai was given for 6 months in a dose 300 mg/day to 63 NIDDM patients aged 40-69 years. The examination covered indices of glycemia, glycosylated hemoglobin, pancreatic insulin secretion, total cholesterol (TC), triglycerides (TG), HDLP and LDLP cholesterol, atherogenic index. RESULTS: Glucobai has improved glycemic profile, reduced insulinemia and body mass index, TC, TG, atherogenic index. Blood levels of HDLP cholesterol elevated by 62%. CONCLUSION: Long-term intake of acarbose improves carbohydrate metabolism and produces a hypolipidemic effect. PMID- 11019431 TI - [Hypolipidemic effects of alisat and lipostabil in patients with diabetes mellitus]. AB - AIM: To study a hypolipidemic action of alisat and lipostabil in patients with non-insulin-dependent diabetes mellitus (NIDDM). MATERIAL AND METHODS: Changes in blood lipids were studied in 121 NIDDM patients aged 36-66 years with compensated or subcompensated carbohydrate metabolism on sugar-reducing therapy. The latter consisted of a 52-week course of alisat (600 mg/day) or lipostabil (900 mg/day) in baseline levels of total cholesterol (TC) under 6.5 mmol/l and above 6.5 mmol/l, respectively. RESULTS: Alisat and lipostal treatments reduced TC levels from 5.4 +/- 25 to 4.77 +/- 0.12 mmol/l and from 7.07 +/- 0.24 to 5.92 +/- 0.30, LDLP cholesterol from 4.0 +/- 0.31 to 2.98 +/- 0.15 mmol/l and 5.54 +/- 0.25 to 4.04 +/- 0.34 mmol/l, respectively. Lipostabil changed LDLP cholesterol and triglycerides from 0.51 +/- 0.05 to 0.33 +/- 0.03 mmol/l and from 2.54 +/- 0.25 to 1.66 +/- 0.15 mmol/l, respectively, while HDLP cholesterol rose from 1.22 +/- 0.10 to 1.55 +/- 0.07 mmol/l. Alisat did not change significantly. CONCLUSION: Adjuvant lipostabil is recommended in combined treatment of NIDDM in marked dyslipidemia, alisat--in moderate dyslipidemia. PMID- 11019432 TI - [Inhalatory corticosteroids in therapy of bronchial asthma]. PMID- 11019433 TI - [Antiviral and regulatory interferon-inducing effects of dipyridamole]. PMID- 11019434 TI - [Reactive oxygen species in progression of chronic myeloid leukemia: prospects of natural antioxidants]. PMID- 11019435 TI - XIII International AIDS Conference highlights. PMID- 11019436 TI - Current issues in antiretroviral and psychiatric therapy for HIV-infected patients. PMID- 11019437 TI - Cervical dysplasia in a nonadherent woman with AIDS. PMID- 11019438 TI - The corrections connection. PMID- 11019439 TI - Inexpensive AIDS drug still reduces HIV transmission from mother to child after 1 year. PMID- 11019440 TI - Adding IL-2 to anti-HIV drugs increases CD4+ cell counts without raising HIV levels. PMID- 11019441 TI - Global initiative to prevent the spread of AIDS. PMID- 11019442 TI - Physician prescribing of sterile injection equipment to prevent HIV infection: time for action. PMID- 11019443 TI - HIV vaccine trials for Africa. PMID- 11019444 TI - Aim at 95% adherence to PIs. PMID- 11019445 TI - Reemergence of plasma viremia after discontinuation of HAART. PMID- 11019446 TI - HIV risk in young men who have sex with men. PMID- 11019447 TI - Severe hypersensitivity reactions following reintroduction of abacavir. PMID- 11019448 TI - Immunization. Update: search for an AIDS vaccine. AB - Identifying a successful HIV preventive vaccine is among the highest research priorities of the US NIH. While therapies for HIV have brought hope to those who are already HIV-infected, stopping the worldwide epidemic will require safe, effective HIV vaccines. Here, we describe scientific and other obstacles to attaining that goal and provide a brief synopsis of clinical trial results, recent preclinical results, and future priorities. PMID- 11019449 TI - Management trends. Resting energy expenditure in the HAART era. AB - HIV infection in the HAART era is often accompanied by nutritional complications, including rapid weight loss and weight gain. Changes in weight are directly related to shifts in energy balance. Understanding the interplay between dietary intake and energy utilization is important for investigating the factors contributing to a person's weight shifts. Total energy use includes resting metabolic rate, or resting energy expenditure (REE), and voluntary activity, both of which are commonly abnormal in persons with HIV infection. There is now evidence that both the level of viral load and the use of HAART affect REE. The role of energy expenditure in lipodystrophy is unknown but is an area of research interest. The current understanding of energy balance in persons with HIV infection is explained here for the health care provider. PMID- 11019451 TI - Robert Wood Johnson Faith in Action funds. PMID- 11019450 TI - HIV Emergency Assistance Program Funding. PMID- 11019452 TI - Maxim and Roche collaborate for development of hepatitis C drug. PMID- 11019453 TI - Psychosocial impact of the lipodystrophy syndrome in HIV infection. AB - Lipodystrophy is a poorly understood condition associated with antiretroviral therapy in HIV infection. The symptoms may include some combination of central fat accumulation, peripheral fat depletion, and metabolic disturbance. A qualitative survey of 33 HIV-infected heterosexual women and gay men with lipodystrophy assessed psychosocial impact and effect on quality of life. Dominant themes included erosion of self-image and self-esteem, problems in social and sexual relations, threats to locus of control, forced HIV disclosure, and demoralization and depression. Another theme was clinicians' minimization of the importance of lipodystrophy. Further research is required to fully understand the psychosocial impact of lipodystrophy and to develop strategies that help individuals cope. PMID- 11019454 TI - Clinical research with topical microbicides as a potential HIV prevention method. AB - An overview of the different phases in microbicide research is given with special attention to nonoxynol-9, the only ingredient that has been tested in phase III trials so far. The methodological and ethical challenges in conducting phase III studies are described. These include study population, compliance data, standardization, therapy for HIV-positive women, and informed consent. Despite these challenges, phase III trials are feasible and should be initiated now. PMID- 11019455 TI - The sustainability transition: a new challenge. PMID- 11019456 TI - Environmental lead exposure: a public health problem of global dimensions. AB - Lead is the most abundant of the heavy metals in the Earth's crust. It has been used since prehistoric times, and has become widely distributed and mobilized in the environment. Exposure to and uptake of this non-essential element have consequently increased. Both occupational and environmental exposures to lead remain a serious problem in many developing and industrializing countries, as well as in some developed countries. In most developed countries, however, introduction of lead into the human environment has decreased in recent years, largely due to public health campaigns and a decline in its commercial usage, particularly in petrol. Acute lead poisoning has become rare in such countries, but chronic exposure to low levels of the metal is still a public health issue, especially among some minorities and socioeconomically disadvantaged groups. In developing countries, awareness of the public health impact of exposure to lead is growing but relatively few of these countries have introduced policies and regulations for significantly combating the problem. This article reviews the nature and importance of environmental exposure to lead in developing and developed countries, outlining past actions, and indicating requirements for future policy responses and interventions. PMID- 11019457 TI - Indoor air pollution in developing countries: a major environmental and public health challenge. AB - Around 50% of people, almost all in developing countries, rely on coal and biomass in the form of wood, dung and crop residues for domestic energy. These materials are typically burnt in simple stoves with very incomplete combustion. Consequently, women and young children are exposed to high levels of indoor air pollution every day. There is consistent evidence that indoor air pollution increases the risk of chronic obstructive pulmonary disease and of acute respiratory infections in childhood, the most important cause of death among children under 5 years of age in developing countries. Evidence also exists of associations with low birth weight, increased infant and perinatal mortality, pulmonary tuberculosis, nasopharyngeal and laryngeal cancer, cataract, and, specifically in respect of the use of coal, with lung cancer. Conflicting evidence exists with regard to asthma. All studies are observational and very few have measured exposure directly, while a substantial proportion have not dealt with confounding. As a result, risk estimates are poorly quantified and may be biased. Exposure to indoor air pollution may be responsible for nearly 2 million excess deaths in developing countries and for some 4% of the global burden of disease. Indoor air pollution is a major global public health threat requiring greatly increased efforts in the areas of research and policy-making. Research on its health effects should be strengthened, particularly in relation to tuberculosis and acute lower respiratory infections. A more systematic approach to the development and evaluation of interventions is desirable, with clearer recognition of the interrelationships between poverty and dependence on polluting fuels. PMID- 11019458 TI - Contamination of drinking-water by arsenic in Bangladesh: a public health emergency. AB - The contamination of groundwater by arsenic in Bangladesh is the largest poisoning of a population in history, with millions of people exposed. This paper describes the history of the discovery of arsenic in drinking-water in Bangladesh and recommends intervention strategies. Tube-wells were installed to provide "pure water" to prevent morbidity and mortality from gastrointestinal disease. The water from the millions of tube-wells that were installed was not tested for arsenic contamination. Studies in other countries where the population has had long-term exposure to arsenic in groundwater indicate that 1 in 10 people who drink water containing 500 micrograms of arsenic per litre may ultimately die from cancers caused by arsenic, including lung, bladder and skin cancers. The rapid allocation of funding and prompt expansion of current interventions to address this contamination should be facilitated. The fundamental intervention is the identification and provision of arsenic-free drinking water. Arsenic is rapidly excreted in urine, and for early or mild cases, no specific treatment is required. Community education and participation are essential to ensure that interventions are successful; these should be coupled with follow-up monitoring to confirm that exposure has ended. Taken together with the discovery of arsenic in groundwater in other countries, the experience in Bangladesh shows that groundwater sources throughout the world that are used for drinking-water should be tested for arsenic. PMID- 11019459 TI - Guidelines for the microbiological quality of treated wastewater used in agriculture: recommendations for revising WHO guidelines. AB - Three different approaches for establishing guidelines for the microbiological quality of treated wastewater that is reused for agriculture are reviewed. These approaches have different objectives as their outcomes: the absence of faecal indicator organisms in the wastewater, the absence of a measurable excess of cases of enteric disease in the exposed population and a model-generated estimated risk below a defined acceptable risk. If the second approach (using empirical epidemiological studies supplemented by microbiological studies of the transmission of pathogens) is used in conjunction with the third approach (using a model-based quantitative risk assessment for selected pathogens) a powerful tool is produced that aids the development of regulations. This combined approach is more cost-effective than the first approach and adequately protects public health. The guideline limit for faecal coliform bacteria in unrestricted irrigation (< or = 1000 faecal coliform bacteria/ 100 ml) is valid, but for restricted irrigation < or = 10(5) faecal coliform bacteria/100 ml is recommended when adult farmworkers are exposed to spray irrigation. A limit of < or = 10(3) faecal coliform bacteria/100 ml is recommended if flood irrigation is used or children are exposed. The guideline limit for nematode eggs for both types of irrigation is adequate except when conditions favour the survival of nematode eggs and where children are exposed; in these cases it should be reduced from < or = 1 egg/l to < or = 0.1 egg/l. PMID- 11019460 TI - The urban environment and health in a world of increasing globalization: issues for developing countries. AB - Urban living is the keystone of modern human ecology. Cities have multiplied and expanded rapidly worldwide over the past two centuries. Cities are sources of creativity and technology, and they are the engines for economic growth. However, they are also sources of poverty, inequality, and health hazards from the environment. Urban populations have long been incubators and gateways for infectious diseases. The early industrializing period of unplanned growth and laissez-faire economic activity in cities in industrialized countries has been superseded by the rise of collective management of the urban environment. This occurred in response to environmental blight, increasing literacy, the development of democratic government, and the collective accrual of wealth. In many low-income countries, this process is being slowed by the pressures and priorities of economic globalization. Beyond the traditional risks of diarrhoeal disease and respiratory infections in the urban poor and the adaptation of various vector-borne infections to urbanization, the urban environment poses various physicochemical hazards. These include exposure to lead, air pollution, traffic hazards, and the "urban heat island" amplification of heatwaves. As the number of urban consumers and their material expectations rise and as the use of fossil fuels increases, cities contribute to the large-scale pressures on the biosphere including climate change. We must develop policies that ameliorate the existing, and usually unequally distributed, urban environmental health hazards and larger-scale environmental problems. PMID- 11019461 TI - El Nino and human health. AB - The El Nino-Southern Oscillation (ENSO) is the best known example of quasi periodic natural climate variability on the interannual time scale. It comprises changes in sea temperature in the Pacific Ocean (El Nino) and changes in atmospheric pressure across the Pacific Basin (the Southern Oscillation), together with resultant effects on world weather. El Nino events occur at intervals of 2-7 years. In certain countries around the Pacific and beyond, El Nino is associated with extreme weather conditions that can cause floods and drought. Globally it is linked to an increased impact of natural disasters. There is evidence that ENSO is associated with a heightened risk of certain vector borne diseases in specific geographical areas where weather patterns are linked with the ENSO cycle and disease control is limited. This is particularly true for malaria, but associations are also suggested in respect of epidemics of other mosquito-borne and rodent-borne diseases that can be triggered by extreme weather conditions. Seasonal climate forecasts, predicting the likelihood of weather patterns several months in advance, can be used to provide early indicators of epidemic risk, particularly for malaria. Interdisciplinary research and cooperation are required in order to reduce vulnerability to climate variability and weather extremes. PMID- 11019462 TI - Climate change and vector-borne diseases: a regional analysis. AB - Current evidence suggests that inter-annual and inter-decadal climate variability have a direct influence on the epidemiology of vector-borne diseases. This evidence has been assessed at the continental level in order to determine the possible consequences of the expected future climate change. By 2100 it is estimated that average global temperatures will have risen by 1.0-3.5 degrees C, increasing the likelihood of many vector-borne diseases in new areas. The greatest effect of climate change on transmission is likely to be observed at the extremes of the range of temperatures at which transmission occurs. For many diseases these lie in the range 14-18 degrees C at the lower end and about 35-40 degrees C at the upper end. Malaria and dengue fever are among the most important vector-borne diseases in the tropics and subtropics; Lyme disease is the most common vector-borne disease in the USA and Europe. Encephalitis is also becoming a public health concern. Health risks due to climatic changes will differ between countries that have developed health infrastructures and those that do not. Human settlement patterns in the different regions will influence disease trends. While 70% of the population in South America is urbanized, the proportion in sub Saharan Africa is less than 45%. Climatic anomalies associated with the El Nino Southern Oscillation phenomenon and resulting in drought and floods are expected to increase in frequency and intensity. They have been linked to outbreaks of malaria in Africa, Asia and South America. Climate change has far-reaching consequences and touches on all life-support systems. It is therefore a factor that should be placed high among those that affect human health and survival. PMID- 11019463 TI - Protecting human health in a changing world: the role of social and economic development. AB - The biological and physical environment of the planet is changing at an unprecedented rate as a result of human activity, and these changes may have an enormous impact on human health. One of the goals of human development is to protect health in the face of rapid environmental change, but we often fail to do this. The aim in this paper is to distinguish between socioeconomic aspects of development that are likely to be protective and those that are likely to increase vulnerability (the capacity for loss resulting from environmental change). Examples include climate change in the Pacific. We conclude that protecting human health in a changing world requires us to take steps to minimize harmful change wherever possible, and at the same time to be prepared for surprises. The goals of mitigation (reducing or preventing change) and adaptation (response to change) are not mutually exclusive. In fact, steps to make populations more resilient in the face of change are often similar to those that are needed to lighten the load on the environment. We need social policies that convert economic growth into human development. Wider application of sustainable development concepts is part of the solution. In particular, there is a need to promote health as an essential asset of poor and vulnerable populations. It is their key to productivity and to surviving shocks; it is also the key to achieving broader development goals such as universal education. For these reasons it is in the interests of all sectors--economic, social and environmental -to play their particular roles in protecting and improving health. PMID- 11019464 TI - Environmental health--for the rich or for all? PMID- 11019465 TI - Paracelsus and Goethe: founding fathers of environmental health. PMID- 11019466 TI - Developing an immunization safety surveillance system in the Philippines. PMID- 11019467 TI - Adverse events monitoring as a routine component of vaccine clinical trials: evidence from the WHO Vaccine Trial Registry. PMID- 11019468 TI - G8 countries set priorities for infectious diseases but fail to make progress on debt relief. PMID- 11019469 TI - Nelson Mandela calls for unity at the XIIIth International AIDS Conference in Durban, South Africa. PMID- 11019470 TI - Community surveys to identify health priorities. PMID- 11019471 TI - Plasma HIV-1 RNA quantitation in HIV infected adult Zimbabweans. AB - OBJECTIVES: To determine HIV-1 RNA levels in plasma of HIV infected Zimbabwean adults and to correlate these with CD4+ cell counts. DESIGN: Prospective observational study. SETTING: Specialist Immunology Laboratory, Harare, Zimbabwe. SUBJECTS: 49 consecutive HIV infected patients. MAIN OUTCOME MEASURES: CD4+ cell counts and plasma HIV-1 RNA levels. RESULTS: Based on the CDC HIV classification system five patients were in early stage infection, 15 in middle, 17 in late and 12 in advanced stage of infection. The median CD4+ cell count was 165/microL (Q1 = 4; Q3 = 866). The median HIV-1 RNA levels were 73,097 RNA copies/ml (Q1 = 753; Q3 = 750,000). Eighty percent of the patients had HIV-1 RNA levels > 10,000 copies/ml and 49% had > 100,000 RNA copies/ml. Median viral loads among the four categories of infection were found to be highly significant (p < 0.001). The correlation between CD4+ cell counts and HIV-1 RNA copies for the 49 patients was highly significant (r = -0.76; p < 0.001). CONCLUSIONS: High treatment HIV-1 RNA levels were determined in the majority of patients. A highly significant correlation was obtained between plasma HIV-1 RNA levels and CD4+ cell counts. PMID- 11019472 TI - Post natal maternal morbidity patterns in mothers delivering in Gweru City (Midlands province). AB - OBJECTIVES: To determine morbid conditions suffered by mothers during the first six weeks post delivery and risk factors associated with them in Gweru district. DESIGN: A cross sectional survey. SETTING: Gweru district hospital, Monomotapa, Mkoba 1 and Mkoba polyclinic. SUBJECTS: A consecutive sample of 201 mothers residing in Gweru district who delivered in these facilities during the month of June 1997. MAIN OUTCOME MEASURES: Morbid conditions, magnitude and the risk factors. RESULTS: During follow up checks on recruited mothers, turn up rates were 82% and 63% at two and six weeks respectively. Fifty eight percent (58%) of the mothers reported at least one morbid condition within the first 24 hours post delivery and pain was a dominant feature. At two weeks, 56% of those who turned up predominantly presented with sepsis in areas of the reproductive tract. The figure fell to 35% among mothers who turned up at six weeks and a mixture of pain and sepsis were the predominant complaints. Episiotomies caused a lot of pain post operatively and the risk of subsequent sepsis was high, (OR: 9; 95% CI: 1.16 < OR < 69.7; p = 0.020). Statistically significant associations were found between backache and multiparity (OR: 1.89; 95% CI: 1 < OR < 3.4; p = 0.040) and also between Caesarian section and some morbid conditions (OR: 4.14; p = 0.002; 95% CI 2.05 < OR < 10.91). The prevalence rate of HIV was 29.4%. HIV positivity was associated with marriage below the age of 19 years (OR: 2.4; 95% CI 1.21 < OR < 3.8; p = 0.024). There was no association between HIV serostatus and maternal morbidity. Use of traditional medication during pregnancy was reported by 42% of mothers, but it did not have any immediately observable intrapartum of post partum effects. Similarly the place of delivery was not associated with post natal maternal morbidity. CONCLUSION: The first two weeks post delivery were the most critical for the mother in terms of post natal maternal morbidity. Sepsis associated pain was the predominant condition. To improve the effectiveness of post natal care, a review within the first two weeks post delivery is an essential intervention, in addition to the routine six weeks check. PMID- 11019473 TI - Abortion: attitudes and perceptions of health professionals in Zimbabwe. AB - OBJECTIVE: To determine the attitudes of professional health workers (doctors, nurses, matrons, social workers and hospital administrators) to medically supervised abortion. DESIGN: Cross sectional study. SETTING: Randomly selected health institutions--urban as well as rural in the eight provinces of Zimbabwe. SUBJECTS: Males and females--age range 18 to 70 years. MAIN OUTCOME MEASURES: Perception of the problem of abortion by health professionals, their knowledge of the present abortion law and desire for change. RESULTS: The majority of doctors were supportive of medically supervised abortion (61.2%) while the nurses were divided 43.2% for and 42.0% against but 14.8% were undecided. The administrators and social workers were supportive. Of the doctors 75% felt that the present abortion law was restrictive and 55.6% supported change. All health professionals agreed that the majority of women who present for abortion treatment are single. The surprising finding was that it is knowledge of the dire complications of unsafe abortion that determines one's attitude to abortion rather than religion. CONCLUSION: By increasing single women's and adolescents' access to family planning services the incidence of unintended pregnancies which result in unsafe abortion with life threatening complications will be reduced. The present restrictive abortion laws which foster backstreet unsafe abortion need to be revised. PMID- 11019474 TI - Patient acceptance of outpatient treatment for inguinal hernia in Jos, Nigeria. AB - BACKGROUND: Patient acceptability constitutes an important component of outpatient treatment for hernias. It is essential that patients are properly selected so that only those who need admission are admitted and cared for with the limited available resources. It is also equally important that those who qualify for outpatient care are adequately informed that there is no added risk attributable to outpatient treatment and that they are free to accept or reject such treatment. OBJECTIVE: To determine the acceptability of day care treatment for inguinal hernia in our environment. DESIGN: Prospective cross sectional study. SETTING: Jos University Teaching Hospital. SUBJECTS: 121 selected patients were randomized to undergo elective inguinal hernia repair either as outpatients (61) or inpatients (60). MAIN OUTCOME MEASURES: Six weeks after operation, an assessment of patients' opinion was made as to their preferred method. RESULTS: 52 of 61 day cases and 24 of 60 inpatients preferred outpatient treatment, while two daycare patients and 36 inpatients preferred hospital admission (p < 0.001). Altogether, acceptability rate for outpatient treatment was 68.6%. CONCLUSION: in carefully selected and adequately informed patients, outpatient elective inguinal hernia repair is readily acceptable. PMID- 11019475 TI - Non-pharmacological termination of a supraventricular tachycardia. AB - Patients with re-entrant supraventricular tachycardias frequently attend casualty departments for termination of their rhythm. With the advent of adenosine in the physician's pharmaceutical armoury, the ease and effectiveness of vagal stimulation is being forgotten. PMID- 11019476 TI - Low CD4 count in HIV negative malaria cases and normal CD4 count in HIV positive and malaria negative patients. PMID- 11019478 TI - Listening is everybody's business. PMID- 11019477 TI - Hypertension in pregnancy--2--eclampsia. AB - Eclampsia is a grand mal convulsion associated with pregnancy-induced hypertension. It is caused by cerebral hypoxia from intense vasospasm combined with cerebral oedema. CT scans show cerebral ischaemia from thrombosis and oedema. PMID- 11019479 TI - Letter across the Pacific. PMID- 11019480 TI - Metapreferences, preferences and suicide. PMID- 11019481 TI - Season and suicide: recent findings from Italy. AB - Seasonal asymmetry in yearly suicide occurrence is a long-observed phenomenon in psychiatric, suicidological, and sociological research, and the effects of seasonal factors on suicidal behavior have been the focus of a number of earlier studies. Taking into account limitations of data and methods, these studies have generally favored interpretations based on psychosocial factors. Recent studies have challenged the widely held notion that seasonal effects on suicide are a unitary phenomenon, not influenced by age, gender, or circumstances of the act. In particular, the seasonal occurrence of suicides has been found to differ significantly between the young and the elderly, and differences have also been found between male and female cycles of occurrence. Suicides using violent methods have been shown to follow clearer seasonal patterns than suicides by less violent methods (such as drug or gas poisoning), possibly reflecting the greater impulsive component involved in the choice of a violent lethal means. In this paper, findings from Italy are used to illustrate the clinical implications of studies into the topic of season and suicide, with the aim of developing more effective preventative strategies. PMID- 11019482 TI - An exercise in improving suicide reporting in print media. AB - This study was conducted to support the publication of guidelines for media reporting on suicide. First, quantitative and qualitative aspects of suicide reporting in Swiss print media were surveyed over a time span of 8 months. The results were presented at a national press conference, and written guidelines for suicide reporting were sent out to all newspaper editors. The results of the survey and the guidelines were discussed in a personal meeting with the Editor-in Chief of the main tabloid. After the publication of the guidelines a second, identical survey was conducted. The main variables regarding frequency, form, and content of the newspaper reports before and after the press conference were compared. The number of articles, on the one hand, increased over the 3 years between the first and second survey, but the quality of reporting clearly improved on the other. The personal contact with the editor of the tabloid was probably the most effective means of intervention. PMID- 11019483 TI - A model for analyzing suicide prevention. AB - This study monitored the evolution of psychologists' (n = 31) conceptions of suicide prevention over the 9-year course of the National Suicide Prevention Project in Finland and assessed the feasibility of the theoretical model for analyzing suicide prevention developed in earlier studies [Upanne, 1999a,b]. The study was formulated as a retrospective self-assessment where participants compared their earlier descriptions of suicide prevention with their current views. The changes in conceptions were analyzed and interpreted using both the model and the explanations given by the subjects themselves. The analysis proved the model to be a useful framework for revealing the essential features of prevention. The results showed that the freely-formulated ideas on prevention were more comprehensive than those evolved in practical work. Compared to the earlier findings, the conceptions among the group had shifted toward emphasizing a curative approach and the significance of individual risk factors. In particular, greater priority was focused on the acute suicide risk phase as a preventive target. Nonetheless, the overall structure of prevention ideology remained comprehensive and multifactorial, stressing multistage influencing. Promotive aims (protective factors) also remained part of the prevention paradigm. Practical working experiences enhanced the psychologists' sense of the difficulties of suicide prevention as well as their criticism and feeling of powerlessness. PMID- 11019484 TI - Telephone crisis intervention skills: a simulated caller paradigm. AB - Counseling skills were evaluated in a telephone counseling service for Vietnam veterans. Thirty simulated crisis calls were made to telephone counselors by experienced clinical psychologists. The counselors' responses were rated on dimensions that indexed general counseling skills, knowledge of veteran needs, and provision of appropriate advice. Whereas the majority of callers demonstrated adequate counseling skills, many lacked knowledge of veterans' posttraumatic stress, common veteran terminology, and the nature of veterans' experiences. The findings suggest that telephone counseling services that target specific populations should employ comprehensive training to ensure that counselors possess adequate and relevant knowledge about callers and their presenting problems. The simulated caller paradigm appears to be an effective paradigm for training and evaluation of telephone counselors. PMID- 11019485 TI - Intraseasonal pattern in the time distribution of suicides. PMID- 11019486 TI - Genetic and molecular pathogenesis of hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterized by vascular dysplasia and hemorrhage. The pathogenesis regarding heterogeneity of vascular malformations in patients with HHT has been obscure, although it has become possible to partially explain the pathogenesis from the identification of two distinct genes, endoglin and ALK-1. Endoglin and ALK-1 are type III and type I TGF-beta receptors, respectively, and are exclusively expressed on vascular endothelial cells. Binding of TGF-beta to the type II TGF beta receptor on endothelial cells, which is accelerated in the presence of endoglin, phosphorylates type I TGF-beta receptors, ALK-5 and ALK-1, and phosphorylated ALK-5 and ALK-1 activate the downstream proteins Smad2/3 and Smad1/5, respectively. These activated Smad proteins dissociate from the type I TGF-beta receptor, bind to Smad4, and enter the nucleus to transmit TGF-beta signaling by regulating transcription from specific gene promoters involved in angiogenesis. Therefore, a balance between these two signaling pathways via ALK-5 and ALK-1 plays an important role in determining the properties of endothelial cells during angiogenesis. Mutations of endoglin and ALK-1 genes are genetic pathogenesis of HHT type 1 and HHT type 2, respectively. To date, at least 29 and 17 different kinds of mutations in endoglin and ALK-1, respectively, have been found, including missense, nonsense, frameshift, and deletion mutations. The precise mechanisms of vascular abnormalities elicited by these mutations observed in HHT patients are still uncertain, although elucidation of the mechanism of intracellular signal transduction and the change in targeted gene expressions using mutant recombinant endoglin or ALK-1 proteins and knockout mice will enable us to understand the genetic and molecular pathogenesis of HHT and to effectively treat patients with HHT. PMID- 11019487 TI - Current status of surgical treatment for hepatocellular carcinoma. AB - The therapeutic modalities for hepatocellular carcinoma (HCC) have diversified during the past decades, and in particular, the treatment has mainly been concentrated on small HCC of 3 cm or less. Therefore, it has become very important for surgeons to be able to identify the clinical indications for liver resection in HCC. We reviewed data on liver resection for HCC using the National Primary Hepatic Cancer follow-up survey report of the Liver Cancer Study Group of Japan, indicating problem associated with liver resection for HCC. As a result, the indications of surgical treatment for HCC are as follows: (1) In patients with HCC of 3 cm or less in diameter, a solitary HCC indicates liver resection. However, priority should be given to medical treatment such as percutaneous transhepatic ethanol injection therapy, microwave coagulative necrosis therapy for multiple HCC and patients with clinical states II or III. (2) HCC between 3 cm and 5 cm in diameter is a good indicator for liver resection. Solitary HCC indicates liver resection as a first choice. (3) HCC greater than 5 cm diameter eter and cases with tumor stages II or III indicate liver resection. (4) There are limits to treating HCC with tumor thrombus in the second branch of the portal vein (Vp2) and tumor thrombus in the first branch, the trunk of the portal vein or in a branch on the opposite side (Vp3) only by liver resection. (5) HCC with tumor thrombus in the right, middle or left hepatic vein trunk, posterior inferior hepatic vein trunk or short hepatic vein (Vv2) and with tumor thrombus in the inferior vena cava (Vv3) are indicators for liver resection. A limited resection according to Glissons structure and with a negative surgical margin can be performed in HCC of 5 cm or less, however an extended resection is required for HCC greater than 5 cm. Furthermore, the extent of liver resection should be considered according to the hepatitis virus. Finally, it was emphasized that effective measures against the postoperative recurrence was essential in order to improve the outcome of HCC. PMID- 11019488 TI - Molecular mechanisms of fragile X syndrome. AB - Fragile X syndrome is the most common form of inherited mental retardation Mutations which abolish expression of an X-linked gene, FMR1, result in pathogenesis of the disease. FMR1 encodes a cytoplasmic RNA-binding protein which interacts with two autosomal homologs, FXR1 and FXR2. These proteins are highly expressed in neurons. In addition, the FMR1/FXR proteins are associated with ribosomes. Given their RNA-binding activity and association with ribosomes, these proteins are hypothesized to bind to specific RNAs and regulate their expression at translational levels in a manner critical for correct development of neurons. Much progress has been made in FMR1 research over the past several years, but little light has yet to be shed on the physiological function of these proteins. It will be critical to define the biochemical properties of these proteins, and identify potential downstream targets to clarify the molecular mechanisms underlying the potential roles of these proteins in translation. A basic understanding of the function of this new family of RNA-binding proteins should then allow us to begin to address the question of how the lack of FMR1 expression leads to symptoms in fragile X syndrome. PMID- 11019489 TI - Multiple endocrine neoplasia type 1: from bedside to benchside. AB - Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterized by the combined occurrence of parathyroid, pancreatic endocrine, and anterior pituitary tumors. MEN1 has two characteristics; a hormone excess and a sometimes lethal outcome due to malignant tumors. The recent identification of the MEN1 gene has opened the door to a much deeper understanding of this syndrome. Germline MEN1 mutations have been identified in most MEN1 families. They were not found, however, in families with familial pituitary tumors. Thus, studies with the MEN1 gene helped to establish that mutation of some other gene(s) is likely causative of the MEN1 phenocopy. These recent advances provide for the identification of mutant MEN1 gene carriers who are at a high risk of developing MEN1. The protein encoded by the MEN1 gene has been shown to function in the regulation of JunD-activated transcription but much still remains to be elucidated. PMID- 11019490 TI - Secondary hyperparathyroidism and phosphate sensing in parathyroid glands. AB - Retention of inorganic phosphate (Pi) and associated hyperphosphatemia are important development of hyperparathyroidism secondary to renal failure. The beneficial effect of a low-Pi diet in the prevention of hyperparathyroidism can be attributed to the decrease in PTH secretion. This effect of Pi may be mediated by specific molecules in the parathyroid cell membrane. A complementary DNA encoding a Na(+)-Pi co-transporter, termed rat PiT-1, has been isolated from rat parathyroid. The amount of PiT-1 mRNA in the parathyroid is controlled by vitamin D and dietary Pi, which are the most important regulators of PTH secretion. The parathyroid Pi transporter may mediate the effects of extracellular Pi and PTH secretion in secondary hyperparathyroidism. In this study, we focus on the function of Na/Pi co-transporters in the parathyroid glands as inorganic Pi sensor. PMID- 11019491 TI - Practice of electroconvulsive therapy at University Hospital, the University of Tokushima School of Medicine from 1975 to 1997. AB - BACKGROUND: In Western nations the effect of ECT has been re-evaluated since the 1970s, while reports on ECT are few in Japan. METHODS: The sample included 3,067 patients admitted to Tokushima University Hospital between 1975 and 1997. Hospital charts were reviewed retrospectively for ECT. RESULTS: ECT was carried out on 6.03% of all subjects; 11.97% of patients with schizophrenia, 4.88% with manic depressive psychosis, 7.44% with atypical psychosis and 3.27% with psychogenic reactions. The remission rate from ECT was 68.11% of all subjects; 59.85% with schizophrenia, 100% with manic depressive psychosis, 100% with atypical psychosis and 92.0% with psychogenic reactions. Patients averaged 10.26 treatments. ECT had been administered mainly to patients who had responded poorly to pharmacotherapy and to patients who required rapid improvement of life threatening symptoms. ECT was highly effective for symptoms as excitement, suicidal tendencies and stupor. Side effects were claimed by 36.77% of patients. CONCLUSIONS: ECT is suggested to be a useful therapeutic modality in current psychiatric practice. PMID- 11019492 TI - Effect of amino acid mixtures on nasal allergic responses induced by toluene diisocyanate in mice. AB - We studied the effect of various amino acid mixtures on nasal allergy induced by the intranasal application of toluene diisocyanate (TDI) in mice. In Experiment 1 (Exp. 1), mice were fed a 25% casein, soy protein isolate (SPI), egg white protein (EW) or gluten diet. In Experiment 2 (Exp. 2), mice were fed a 25% amino acid mixture diets patterned after casein (AA-casein), SPI (AA-SPI), EW protein (AA-EW) or gluten (AA-gluten). In Experiment 3 3 (Exp. 3) we modified the glutamine/glutamic acid (Gln/Glu) concentrations in the amino acid mixtures. Mice were fed a 25% AA-SPI, low Gln/Glu AA-SPI (LG-AA-SPI), AA-EW or high Gln/Glu AA EW (HG-AA-EW) diet. At the 5th week, mice were divided into sensitized (sen-) and non-sensitized (ns-) groups. The mice in sensitized groups were treated with two courses of intranasal application of toluene diisocyanate (TDI) in ethyl acetate for 5 consecutive days, separated by 9 days rest. The non-sensitized groups of mice were treated with a vehicle. Nine days after the second sensitization, all mice were provoked by TDI. Nasal responses and serum IgE concentration were studied. The findings of Exp. 1 showed that the sen-EW group exhibited a lower body weight gain, higher nasal symptom score and higher IgE concentration than the other sensitized groups. The findings dings of Exp. 2 showed that the sen-EW group had a lower body weight gain, higher nasal symptom score and higher IgE concentration than the other sensitized groups. In Exp. 3, the AA-EW group showed a higher total nasal score and IgE concentration than the HG-AA-EW group, however, the findings of LG-AA-SPI and AA-SPI were similar. These findings demonstrated that amino acid mixtures affect nasal allergy induced by the intranasal application of TDI in mice. PMID- 11019493 TI - Comparison between short-term food restriction and exercise on whole body glucose disposal in high-fat rats. AB - High-fat diets induce whole-body insulin resistance. The aim of this study was to compare effect of two interventions: 3-day food restriction (66% of ad libitum fed) and 3-day exercise training (voluntary running wheels), on decreased insulin mediated whole body glucose uptake in high-fat fed rats (5 mo old) using the hyperinsulinemic euglycemic clamp procedure. The control group was maintained on rat chow alone. After high-fat feeding for 2 wk, insulin-stimulated whole body glucose utilization was significantly decreased by 26%. The exercise training was more effective than food restriction in lowering plasma concentrations of insulin and triacylglycerol and tissue concentrations of triacylglycerol in soleus muscles. Diminished whole-body glucose uptake resulting from high-fat feeding was reversed completely by exercise training, but only partially by food restriction. The time course of starvation on insulin-stimulated glucose uptake was also observed in high-fat fed rats. Although the extension of starvation time to 48 h resulted in decreased plasma glucose, insulin and triacylglycerol concentrations, whole body glucose uptake did not increase further. These findings suggest that short-term exercise has a higher restorative effect on insulin sensitivity in high-fat fed rats than food restriction, in spite of the same loss in body weight, presumably due in part to improved local lipid availability. PMID- 11019494 TI - An incident involving blood sucking by a tick in a suburb in Japan. AB - We encountered a patient whose blood was sucked by Haemaphysalis longicornis in the suburb of a business city in Tokushima prefecture in Japan. The tick, which had been attached to the lower limb of the patient for one week, measured 10 mm in length. There were no notable objective or subjective findings after the complete extirpation of the tick. The area had not been known in recent history to be a habitat of ticks, and, thus, this case is of importance in terms of predicting future trends of tick-borne diseases in Japan. PMID- 11019495 TI - Ovarian fibrothecoma with massive edema. AB - We report a rare case of ovarian fibrothecoma with massive edema. The patient was a 59-year-old woman with a left ovarian mass measuring 11 x 10 x 7 cm. Magnetic resonance images revealed a solid mass showing unhomogeneous content with predominantly high signal intensity on T2-weighted image. Microscopically, the ovarian mass was composed of a cellular area and an edematous hypocellular area. The latter accounted for more than 75% of the tumor. In the cellular area, spindle-shaped or plump tumor cells were randomly distributed or arranged in a fascicular fashion. These cells contained abundant intracytoplasmic lipid. There was dense collagenous connective tissue in the stroma of the cellular areas. In contrast, in the edematous areas spindle or stellate cells were scattered. Alcian blue stain revealed only a small amount of stromal mucin even in the edematous areas. The microscopic findings were consistent with that of fibrothecoma with massive edema. The present case must be differentiated from massive edema of the ovary and sclerosing stromal tumor of the ovary. Immunohistochemistry was not helpful in distinguishing them. The age of the patient and careful histologic observation are important. PMID- 11019496 TI - Diffuse intervertebral disk calcification in a patient with rheumatoid arthritis. AB - A patient with seronegative rheumatoid arthritis (RA) who presented with intervertebral disk calcification (IDC) of several thoracic and lumbar intervertebral disks in herein described. There was no evidence of any other coexisting diseases such as ochronosis and hemochromatosis, but a remarkable degree of polyclonal hypergammaglobulinemia was observed as a notable finding. Although the appearance of IDC on T1-weighted images on magnetic resonance is controversial, no increased signal intensity was observed in our patient. To the best of our knowledge, this is the first report of IDC in RA. PMID- 11019497 TI - Hepatocellular carcinoma with extension into the right atrium: report of a successful liver resection by hepatic vascular exclusion using cardiopulmonary bypass. AB - We report a successful liver resection using cardiopulmonary bypass with, total hepatic vascular exclusion (THVE) for hepatocellular carcinoma (HCC), with extension into the right atrium. A 61-year-old man with a cirrhotic liver was referred to our department with HCC in the medial segment of the left lobe of the liver, and tumor thrombus extending into the right atrium. During surgery, a left lobe and caudate lobe of the liver were transected leaving the left lobe of the liver connected to the inferior vena cava (IVC) by only the left and middle hepatic trunks, and then the intracaval tumor thrombus and the left lobe of the liver were removed en bloc using cardiopulmonary bypass with total hepatic vascular exclusion (THVE). Cardiac arrest was not performed during THVE, and the patient had an uneventful postoperative course and was discharged from the hospital 2 months following surgery. He died of multiple pulmonary metastases 4 years and 8 months after surgery; however, imaging showed no evidence of recurrence in the remnant liver during that period. In conclusion, by performing dissection of the hepatic parenchyma to the hepatic vein prior to removal of the tumor thrombus, the period of extracorporeal circulation, duration of warm ischemic time to the liver, and intraoperative blood loss were all reduced and a radical operation could be performed safely without scattering tumor cells during extirpation of the tumor. PMID- 11019498 TI - Human pulmonary dirofilariasis presenting as a small nodule with a cavity. AB - A 73-year-old woman had a 4-year history of lung fibrosis from collagen vascular disease. She presented with a complaint of dry cough. A chest radiograph showed a 2-cm solitary pulmonary nodule with a small cavity in the right lower lobe. Preoperatively, we performed computed tomography of the chest and measured tumor markers. Video-assisted thoracotomy was performed because we could not rule out lung cancer. Pathologic analysis confirmed the presence of a granuloma with Dirofilaria immitis. In Japan, the incidence of human dirofilariasis has steadily increased and must be considered in the workup of cavitary pulmonary nodules. PMID- 11019499 TI - Hepatopulmonary syndrome-discussion of cardiopulmonary parameters. AB - We report a 70-year-old man with hepatopulmonary syndrome (HPS) in C liver cirrhosis. Hypoxemia worsened markedly, especially on exertion, while the hepatic function was clinically stable. Contrast echocardiography, 99mTc macroaggregated albumin (99mTcMAA) lung scan, and pulmonary angiography were performed. The findings suggested the presence of both intrapulmonary vascular dilatation and substantial right-to-left shunt. The contribution of intrapulmonary vascular abnormalities in patients with severe liver cirrhosis without abnormal chest radiography and spirometry tests when marked hypoxemia is present should be investigated. PMID- 11019500 TI - Care of the sick and the dying: a matter of trust? PMID- 11019501 TI - Family Medicine residents' knowledge and attitudes about end-of-life care. AB - The medical management of end-of-life symptoms, and the psychosocial care of the dying and their families have not been a specific part of the curriculum for undergraduate medical students or residency training programs. The purpose of our research was to assess family medicine residents' knowledge of and attitudes toward care of the dying. All entering (PGY1) and exiting (PGY2) residents of the Dalhousie University Family Medicine Residency Program were given a 50-item survey on end-of-life care. They survey contains two 25-item subscales concerning attitudes/opinions toward end-of-life care, and knowledge about care. Thirty-one of the 33 entering PGY1s 94%) and 26 of the 30 exiting PGY2s (86%) completed the surveys. Overall attitude scores were felt to be high among both groups, with little difference between them. Areas of concern regarding the adequacy of knowledge were found in relation to managing opioid drugs and the symptom of dyspnea. Interventions are now in development to address these issues in the residency program. In an era of subspecialties, the challenge of integrating these areas into the curriculum without creating rotations in specialist palliative care is an issue faced by most family medicine residency programs. PMID- 11019502 TI - The Canadian Palliative Care Education Group. AB - The Canadian Palliative Care Education Group, a committee of the Canadian Society of Palliative Care Physicians, carries out surveys from time to time on the status of palliative care education in Canadian medical schools. We describe the organization of the Canadian Palliative Care Education Group and report on a survey carried out in the 1996-97 academic year. Our data suggest that the emphasis on palliative care varies widely amongst the 16 Canadian medical schools, in concert with the availability of academic staff positions in palliative care. We conclude that the need for palliative care education is supported by rhetoric which remains to be matched by proportionate tangible investment. PMID- 11019503 TI - Frequency of symptom distress and poor prognostic indicators in palliative cancer patients admitted to a tertiary palliative care unit, hospices, and acute care hospitals. AB - The Edmonton Regional Palliative Care Program was established to increase access to palliative care for terminal cancer patients in the region. Inpatient care is delivered, in decreasing order to distress, at the tertiary palliative care unit, by consult teams in acute care facilities, and in hospices. We reviewed the admission data for all patients discharged from the program between November 1, 1997, and October 31, 1998, in order to determine if demographical and clinical variables suggested appropriate use of the three levels of care. Patients admitted to the tertiary palliative care unit were significantly younger than those admitted to acute care of hospices (61 years versus 68 years and 71 years respectively, p < 0.0001), had a higher frequency of positive screening for alcoholism in the CAGE questionnaire (27% versus 16% and 14% respectively, p < 0.0001), and a higher frequency of poor prognostic pain syndromes as compared to acute care admissions (87% versus 65%, p < 0.0001). Overall, frequency of symptoms and severe symptoms was significantly higher in patients admitted to the palliative care unit than those admitted to the other two settings. Our results suggest that patients with demographic and clinical indications of higher distress are more frequently admitted to the tertiary palliative care unit. The clinical tools are useful predictors of utilization that can be used for monitoring health care delivery. PMID- 11019504 TI - Hospital referrals to a hospice: timing of referrals, referrers' expectations, and the nature of referral information. AB - Hospital referrals to Mary Potter Hospice were reviewed prior to the implementation of a new hospital palliative care support service. The hospital palliative care service aims to improve the communication between health professionals in the acute hospital and hospice, and to advise the hospital staff on its management of terminally ill patients. The purpose of this review was to provide baseline information that would assist the hospice with the planning and future evaluation of the new service. Information included the timing of the referrals, the type of referral information provided by hospital doctors, and their expectations of hospice care. One fifth of patients were referred near to death. While medical information was nearly always provided on referral, information on the psychological, spiritual and social dimensions of care was often absent. A third of hospital doctors expressed their expectation of the hospice as "to take over" the patient's care. This suggests the hospital palliative care support service should encourage health professionals to take a more active role in caring for dying patients. The findings are discussed in relation to the goals of the new hospital palliative care support service. PMID- 11019505 TI - An exercise intervention for advanced cancer patients experiencing fatigue: a pilot study. AB - Fatigue is reported by advanced cancer patients to be their most prevalent and distressing symptom. Despite this, few interventions have been developed and tested to manage this debilitating symptom. This paper describes a pilot study undertaken to test the effects of a 28-day exercise intervention on levels of fatigue in advanced cancer patients. All participants were able to increase their activity levels with no increase in reported fatigue. Furthermore, a trend was noted in all patients toward increased quality of life scores and decreased anxiety scores. All participants described a sense of satisfaction in attaining increased activity levels. These preliminary pilot results suggest that patients who initially report the highest levels of fatigue may achieve the largest decrease in fatigue scores. These findings provide support for the suitability of this intervention for the palliative care population and justify the importance of further hypothesis testing. PMID- 11019506 TI - Death education in home hospice care in Japan. AB - In the practice of home hospice care, death education for both patient and family is extremely important, although little information on its usefulness is available. In this study, the effects of death education under home hospice care were analyzed for 16 patients who died at home. Death education for the patient and his/her family was given at least once in each phase of care, and at least four times in total. The acceptance of death by the patients was judged according to the way they spent their remaining time, to their attitudes, and to their hope for a life after death. Fourteen of 15 patients appeared to accept their own death. An autopsy was performed in five of the 16 cases. In one case, the doctor recommended an autopsy to the family; in the other cases, it was performed in accordance with the patient's or family's wish. As the goal of death education in home hospice care is the acceptance of death by both patient and family, our methods of death education appear to be effective. PMID- 11019507 TI - Factors influencing outcomes for family caregivers of persons receiving palliative care: toward an integrated model. PMID- 11019508 TI - Article: Comparison of CMA Joint Statement on resuscitative interventions and New Brunswick Hospital Corporations' policies on end-of-life treatments. Poirier N. J Palliat Care 2000; 16(1): 15-22. PMID- 11019509 TI - [Current topics on classification and nomenclature of bacteria. 1. Preface and schedule of the series]. PMID- 11019510 TI - [Clinical study of infection associated hemophagocytic syndrome]. AB - Hemophagocytic syndrome (HPS) causes pancytopenia, increased blood LDH level, liver dysfunction, DIC, etc. with macrophages appearing in the bone marrow, spleen, lymph nodes, etc. Adult HPS is mostly secondary to various infections, malignant tumors, etc. and sometimes has a serious outcome. Particularly infection associated HPS (IAHS) is triggered by viral, bacterial and fungal infections. The cases of unknown primary disease and suspected IAHS of unidentified pathogenic microorganism are often encountered in the clinical setting. The authors compared IAHS and malignant associated HPS (MAHS) and classified IAHS into viral associated HPS (VAHS), bacterial associated HPS (BAHS) and fungal types to compare the test values based on the test findings at the onset in the HPS cases treated at our Department. The patients consisted of 21 HPS cases, 11 IAHS cases (VAHS 4, BAHS 5, fungal 2) and 10 MAHS cases. Based on the test findings (WBC, Hb, Plt, LDH, ferritin, myelogram, cytokines, [IFN alpha, TNF gamma, IL-6, sIL-2R, M-CSF], adhesion molecules [sICAM-1, sVCAM-1, sELAM-1, sL-selectin]) at the onset, a comparison between IAHS and MAHS and among the IAHS cases classified by pathogenic microorganism was made. In the comparison between IAHS and MAHS, the Hb value was significantly decreased and sIL-2R tended to be increased at the onset in MAHS. When comparing the IAHS cases by pathogenic microorganism, Plt was significantly decreased and sICAM-1 and sVCAM-1 were increased at the onset in the BAHS, The BAHS cases had serious underlying diseases and poor prognosis with high incidence of DIC complications. We are going to accumulate more cases for early diagnosis and treatment of IAHS. PMID- 11019511 TI - [Evaluation of antiretroviral chemotherapy based on the profile of virus isolation from HIV-1 infected individuals]. AB - It is well-known that the biological characteristics of HIV-1 persistently infected in the host have often changed into a rapid growth in vitro, T-cell line tropism and marked syncytium inducing (SI) ability accompanied by the progress of clinical stages from AC to ARC or AIDS. We have developed a follow-up diagnostic test using the clinical markers based on the virus phenotypes mentioned above, and reported that the test was significantly useful for determining the clinical status of HIV infected individuals. Recently, highly active antiretroviral therapy (HAART) was introduced into HIV-1 chemotherapy, and has been reported to be a markedly effective treatment for HIV infected individuals. In this study, we carried out an investigation to see whether the follow-up diagnosis was useful even after introducing the HAART in Japan in 1997. The results by the laboratory diagnosis on 139 HIV infected individuals who were clinically observed over a long period showed that the positive rate of virus isolation and MT-4 cell tropism in the isolates during the two years between 1997 and 1998 were significantly lower than that of the nine years from 1988 to 1996 before the implementation of HAART. In addition, we obtained data that the effect of HAART reflects the biological profiles of virus isolation more than the CD4+ T cell counts. These results suggest that data of clinical examination using virus isolation as a parameter are useful not only for predicting the development of AIDS but also for evaluating the effects of HAART, particularly in patients showing no association between the CD4+ T cell counts and the plasma viral load. PMID- 11019512 TI - [Incidence of amantadine-resistant influenza A (genotype Ser-31-Asn) in nursing homes in Niigata, Japan]. AB - The antiviral agent amantadine specifically inhibit influenza A virus infection, but the emergence of drug-resistant viruses occur more readily with amantadine treatment. In human influenza viruses, single amino acid changes at 4 sites spanning the transmembrane domain of the M2 protein can confer drug resistance. Amantadine was approved for treatment of Parkinson's disease in 1975, and for the influenza A virus infection in November 1998, in Japan. Annual consumption of amantadine for influenza treatment increased suddenly after the approval. According to our previous report, the predominant genotype of resistant virus is the substitution S-31-N in M2 both in vitro and in clinical samples, as in the other reports. Based on the above findings, we focused on single amino acid change at position 31 (genotype S-31-N) and applied polymerase chain-reaction restriction fragment length polymorphism (PCR-RFLP), directly from throat swab samples, by using primers that insert a restriction site for Sca I. With this technique, we surveyed the incidence of amantadine resistant viruses in nursing homes, Niigata, Japan. Thirty-one (22.0%) of 141 PCR positive samples from 8 nursing homes in 1998-99 season showed resistant patterns, and only 6 (19.4%) of them were after the administration of amantadine for treatment. All of these 8 nursing homes used amantadine for Parkinson's disease, but only half of them used the drug for influenza A infection. The incidence of resistant viruses was not significantly different from facilities with amantadine for influenza treatment to those without, 25.5% and 14.0% respectively. The occurrence of outbreaks and sporadic illness in those facilities, with different administration status were observed, but fortunately we could not find any evidence to relate the emergence of resistant viruses to the outbreaks. This is the first report that the resistant influenza viruses already exist in nursing facilities where amantadine was used for not only influenza but also Parkinson's disease in Japan. PMID- 11019513 TI - [Search for Staphylococcus aureus heterogeneously resistant to vancomycin (hetero VRSA) in MRSA strains isolated from clinical samples during 1990s]. AB - Hetero-VRSA was studied in 978 MRSA strains isolated from clinical samples during 1991 to 1998. Although no VRSA was detected, 23 strains (2.4%) were identified as hetero-VRSA by the vancomycin-resistance using MU3 agar plate. The frequency of hetero-VRSA was not increased in the course of time. MIC of the hetero-VRSA to vancomycin and teicoplanin was 1-2 micrograms/ml and 0.5-12 micrograms/ml, respectively. All of the hetero-VRSA strains were confirmed as a heterogeneous strain by a population analysis. Although 43% of the hetero-VRSA strains were coagulase type II, positive for TSST-1, and enterotoxin type C, others were various in the characteristics. In the gene analysis by pulse field gelelectrophoresis (PFGE), 4 sets of 2 strains were found to be identical among the 23 strains but the other 15 strains were genetically different. We speculate that hetero-VRSA strains were generated in 1991 secondary possibly by use of beta lactam antibiotics. PMID- 11019514 TI - [Report of questionnaire survey for methicillin-resistant Staphylococcus aureus and penicillin-resistant Streptococcus pneumoniae in the Kinki District]. AB - An epidemiological investigation for methicillin-resistant Staphylococcus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae (PRSP) was performed at 22 medical institutes in Kinki area by the questionnaire from Kinki Infection Working Group in 1998. The isolation frequencies of MRSA and PRSP, including penicillin-intermediate resistant S. pneumoniae (PISP), were 61.5 and 45.8%, respectively. As compared to the previous data of MRSA in the Kinki area of 1997, the increase of MRSA was recognized. A single infection with MRSA was 49.5% and simultaneous pathogen detected with MRSA was Candida species. On the other hand, a single infection with PRSP including PISP, was 36.4% and simultaneous pathogen with PRSP was Haemophilus influenzae. Vancomycin, teicoplanin, ST and arbekacin showed good activity against MRSA. PRSP and PISP had low activity to ampicillin and erythromycin. PMID- 11019515 TI - [Virucidal activity of disinfectants. Influence of the serum protein upon the virucidal activity of disinfectants]. AB - Five disinfectants were tested for virucidal activity on three DNA viruses and three RNA viruses in the presence or absence of serum protein. Disinfectants of the aldehyde and halogen groups had a virucidal activity on human herpes virus, bovine rhabdo virus, human immunodeficiency virus, human adeno virus, porcine parvo virus, and polio virus. Disinfectants of the invert and amphoteric soap groups, and biganide group had a destructive effect on RNA and DNA viruses possessing an envelope. The presence of serum protein exerted great influence upon the virucidal activity of disinfectants of the invert and amphoteric soap groups. PMID- 11019516 TI - [Comparison of positivity rates for antibodies against measles, rubella, chickenpox and mumps by assays]. AB - Our previous study found mistakes by some doctors in the choice of an assay for determining antibodies in Japan. To compare the positivity rates for antibodies by assays, we measured the antibodies of measles, rubella, chickenpox and mumps from the same sera using such methods as the EIA, HI and CF assays. The subjects were 175 nursing students. The positivity rates for measles, chickenpox and mumps by the EIA assay were 96.6%, 93.7%, and 83.3%, respectively. Those for rubella by the HI and CF assays were 92.0% and 10.1%. The sensitivity rates for measles, chickenpox and mumps by the HI and CF assays, based on the results of the EIA assay, were 75.1%, 102.4% and 69.2% in the HI assay, and 20.6%, 38.7% and 8.0% in the CF assay, respectively. Our previous study showed that the sensitivity of the HI assay for rubella antibody is same as that of the EIA assay in Japan. Currently an EIA assay should be chosen for these antibodies and the HI assay or IAHA assay should be possible selections for rubella and chickenpox. However, international comparison of the cutoff titers for these antibodies should be considered. PMID- 11019517 TI - [Cytomegalovirus (CMV) infection in infants may result intractable stridor]. AB - We found ten cases of human cytomegalovirus (CMV) infection who were intractable stridor. Their symptoms were not improved by the treatment with aminophyllin nor beta stimulants. They were admitted repeatedly complaining of stridor, fever and diarrhea. In two cases, the immunological findings showed a decrease of bacterial sterilizing activity of the neutrophils. Additionally, blood count showed leukocytosis more than 15,000/ul in all cases. Total serum IgE and specific IgE antibodies to many antigens were not elevated. Transaminase was elevated. Chest X p findings of interstitial pneumonia or atelectasis continued for a long time in some cases. Virological examinations revealed high concentrations of specific IgM or CF antibodies against CMV in all cases. CMV DNA in saliva were examined by polymerase chain reaction (PCR) with primer sets for the immediate early (IE) region of CMV and showed positive in seven cases. CMV in bronchoalveolar lavage (BAL) was isolated in two cases, and CMV PCR in BAL was positive in three cases. The sequence of the CMV-PCR products showed almost same sequence except one point mutation in bp 1203. We considered that CMV infections in infants may induce stridor for a long period. PMID- 11019518 TI - [Certain clinical aspects of etiology of internal diseases]. PMID- 11019519 TI - [Apoptosis and myocardial pathology]. PMID- 11019520 TI - [Interactive regimen of chronodiagnosis and biocontrolled chronophysiotherapy in some internal diseases]. PMID- 11019521 TI - [Current outlook on pathogenesis and treatment of chronic cardiac insufficiency]. PMID- 11019522 TI - [Clinical course, diagnosis and treatment of malabsorption syndrome]. PMID- 11019523 TI - [Gastroesophageal reflux disease: new data on the mechanisms of esophageal ulcer lesion healing]. PMID- 11019524 TI - [Permanent and disseminated intravascular coagulation]. PMID- 11019526 TI - [Infectious endocarditis in intravenous drug abusers]. AB - Clinical course of infectious endocarditis (IE) was analysed for 43 intravenous drug abusers. 42 of them had primary IE, one patient--secondary. Acute course and high activity of the disease were registered in 86% of the patients. IE was provoked by Staphylococcus aureus (50%), Staphylococcus epidermidis 920%), Staphylococcus haemolyticus (11%), E. coli (8%), Pseudomonas aeruginosa (2%), Candida albicans (2%), mixed microflora (7%). Vegetations were detected on the tricuspid, mitral and aortic valves (52, 23 and 19%, respectively), on more than one valve (6%). Pneumonia, pleuricy, hydrothorax, enlargement of the liver, spleen, nephritis and anemia were found in 76, 44, 9, 100, 75, 70 and 88% of the patients, respectively. Cardiac failure aggravated the disease in half of the patients, lethality was 18%. Thus, IE in intravenous drug abusers is characterized by a primary form, acute active course, prevalent damage to the tricuspid valve, polyorganic involvement, high lethality. IE cure in such patients is feasible only in adequate antibacterial therapy, timely surgical correction and giving up drug abuse. PMID- 11019525 TI - [Neopterin: new immunological marker of autoimmune rheumatic disease]. AB - Serum neopterin (SN), concentration of soluble (s) TNF-receptors (R) with molecular mass 55 kD and sIL-2R, C-reactive protein (CRP), Willebrand factor antigen (WF Ag) were measured in enzyme immunoassay (EIA) or radioimmunoassay (BRAHMS, Berlin, Germany) in 189 patients with autoimmune rheumatic diseases: 52 patients with systemic lupus erythematosus (SLE), 67 patients with rheumatoid arthritis (RA), 44 patients with polymyositis/dermatomyositis and 26 patients with Wegener granulomatosis. SN appeared elevated in autoimmune rheumatic diseases correlating with the disease activity and concentrations of sTNF R and sIL 2R. Assay for neopterin is clinically essential for examination of the immunopathological process activity, prediction of the outcomes, better knowledge about cytokine synthesis profile in autoimmune rheumatic diseases. PMID- 11019527 TI - [Microcirculatory and hemostatic disturbances in patients with influenza and respiratory infections aggravated with coexisting diseases]. AB - Microcirculatory disorders (MD) and hemostasis (HS) were studied in patients with influenza and acute respiratory viral infection (ARVI). The patients also suffered from ischemic heart disease (IHD), essential hypertension (EH) and diabetes mellitus (DM). Among 241 patients 63.9% were middle-aged and old. 45 patients under 60 years of age without coexisting diseases served control. In acute influenza and ARVI the majority of the postcapillary veins and capillaries were affected with sludge syndrome, there were marked perivascular and vascular changes. Convalescence was accompanied with reduced permeability and intravascular aggregation of erythrocytes, microvessels improved tonicity. HS responded to the acute infections with depression of fibrinolysis, in convalescence platelet aggregation activated. IHD patients had disseminated intravascular red cell aggregation, slowing of the microflow, hypercoagulation. Patients with postmyocardial infarction cardiosclerosis (PIC) had more severe affection of microcirculation and hemostasis in convalescence. In hypertensive patients microcirculation and hemostasis were similar to those with IHD. In diabetics platelet aggregation improved but sludge phenomenon and slow blood flow persisted. Thus, ARVI for IHD, EH DM patients are a risk factor for the disease aggravation. In influenza and ARVI, IHD patients, especially with PIC and EH are contraindicated active physical exercise, intake of dysaggregant drugs is desirable. Diabetics should take drugs improving blood rheology early in acute period of ARVI. PMID- 11019528 TI - [Diagnosis and treatment of obstructive sleep apnea in internal medicine]. AB - 130 inpatients (94 males and 36 females, mean age 51.8 +/- 9.1 years) with suspected obstructive sleep apnoa (OSA) were examined clinically, were questioned and underwent night cardiorespiratory monitoring. A definite OSA diagnosis was made in 83 patients. Sleep continuous positive airway pressure (CPAP) was applied in 40 patients who had moderate and severe sleep respiratory impairment. The treatment was effective in 31(77%) patients who stopped snoring, improved sleep quality and daytime sleepiness. In half of the hypertensive patients elimination of apnoa lowered blood pressure without drugs in day hours. Patients with chronic respiratory insufficiency improved gas exchange. In pulmonary hypertension CPAP therapy reduced pressure in the pulmonary artery. It is concluded that diagnosis and effective individual therapy of most CPAP patients can be provided in therapeutic hospital departments. PMID- 11019529 TI - [Aspects of hemodynamics in thyroid dysfunction]. AB - Quantitative electrocardiography, echocardiography and variation pulsometry were used in examination of 231 patients with thyroid dysfunction of various degree. Changes in the central and pulmonary hemodynamics, clinical course, ECG depended on thyroid and thyrotropic blood activity, sympathetic and parasympathetic components of autonomic nervous system. Thyrotoxicosis patients had hemodynamic disorders of the greater and lesser circulation, hypothyroidism patients had only the former. PMID- 11019530 TI - [Current aspects of differentiated therapy of infectious endocarditis]. AB - 150 patients (90 males and 60 females, mean age 41.9 +/- 3.3 years) with infectious endocarditis (IE) were treated. 26.7 and 71.3% of the patients had primary and secondary IE, respectively. Clinical and laboratory manifestations of the variants indicate that each variant is associated with a certain clinicolaboratory syndrome: acute IE is associated with toxicoseptic syndrome, subacute--with immune unbalance, and thromboembolic complications, chronic--with immunocomplex visceral lesions. Different variants of the course predetermine different therapy. It is recommended to combined active antibacterial therapy with UV radiation of autoblood and hemosorption, hyperbaric oxygenation. Immunocorrective and efferent therapy hold promise in IE treatment. Glucocorticosteroids are effective in immunocomplex disorders. PMID- 11019531 TI - [Efficacy of peristil treatment in primary and secondary duodenal dyskinesias]. AB - Clinical studies of peristil demonstrated that this drug is effective in hypomotor and dysmotor gastroduodenal disturbances leading to reduction of clinical symptoms of primary and secondary duodenodyskinesias. A systemic prokinetic action of peristil is beneficial in patients with duodenodiskinesia combination with gastroesophageal reflux as well as hypomotor constipation syndrome. Also, peristil reduces depressive symptoms and improves quality of life in patients with primary and secondary duodenodyskinesias. PMID- 11019532 TI - [Use of olicard retard in patients with stable angina of low effort in an outpatient setting]. AB - Efficiency of olicard 40 retard monotherapy and its combination with beta blockers and calcium antagonists was studied in 33 IHD patients with effort angina of functional class III. It was established that olicard retard is an effective antianginal drug pharmakinetics of which allows assignment of individual therapy of IHD outpatients. PMID- 11019533 TI - [Current problems of differential diagnosis and treatment of arterial hypertension]. PMID- 11019534 TI - [Diagnosis of the upper gastrointestinal tract diseases in pregnancy]. PMID- 11019535 TI - [Application of peptide bioregulators in gerontology: results and perspectives]. PMID- 11019536 TI - [A malignant course of pheochromocytoma with marked myocardial hypertrophy]. PMID- 11019537 TI - [Surgical treatment of gastroduodenal ulcer]. PMID- 11019538 TI - [Some moral issues in science and research]. PMID- 11019539 TI - [The importance of periodic medical examinations for the promotion of workers' health in the 21st century]. AB - Organization of periodic medical examinations for workers in XXI century should correspond to requirements of swiftly developing production. Successful future of periodic medical examinations necessitates regular disclosure of new aspects of those requirements and long-term planning of their application to working environment. PMID- 11019540 TI - [The structure of occupational morbidity in electric welders]. AB - Among 6 nosologic entities of occupational diseases diagnosed in 323 electric welders of Moscow suburb, the most frequent one was pneumoconiosis caused by welding aerosol, then neurosensory deafness, chronic toxic dust bronchitis and chronic manganese intoxication. Malignancies located especially in larynx and lungs appeared important among concurrent diseases that could be considered occupational. PMID- 11019541 TI - [Prognosis of inhalation-related injuries in accidental release of ammonium and chlorine by the method of dynamic concentration]. AB - The article deals with new approaches to evaluate casualty effect of accidental chemical release exemplified by common and dangerous industrial chemicals ammonium and chlorine. Dynamic concentration method considers effects of high concentrations inducing instant reflex cardiac and respiratory arrest as well as lesions caused by variable toxic doses. PMID- 11019542 TI - [The results of cytogenetic survey in liquidators of the Chernobyl nuclear accident]. AB - The article represents results of long cytogenetic survey among liquidators of Chernobyl accident. The parameters studied are frequency of unstable chromosomal aberrations and frequency of symmetrical translocations. The authors show that, in spite of long term after irradiation, average frequency of cells with dicentrics and centric fusions (unstable chromosomal aberrations) remains relatively high. Discussion includes possible use of cytogenetic methods in reconstruction of absorbed radiation doses in many years after exposure. PMID- 11019543 TI - [The combined action of protein-containing dust and sulfur dioxide (experimental studies)]. AB - The authors obtained new concepts on immune and biochemical response conditioned by character and circumstances of combined biologic and chemical agents. The article covers role of chemical technogenic pollution promoting transfer of compensated adaptation into decompensation phase with sensibilization or allergy in some cases and with toxic allergic reactions in others. The calculated values include a generalized parameter that, if changed over preset level, could be a risk criterion for the certain diseases. PMID- 11019544 TI - [Chemical contents of dust released by electric enterprises]. AB - Dust in electric lamp plant appeared to contain maximal levels of Sb, Cd, Hg, W, Pb and Sn, that in plant producing light sources and electric vacuum glass--Cu, B, Ag and Pb, that in semiconductor industry and power electronic enterprises- Cd, Mo, Cu, Pb, Ag and Zn, that in cable plant--Cu, Sn, Pb, Sb, Cd. PMID- 11019545 TI - [Aspects of the heat status in the pilot during a continuous flight]. AB - Pilots wearing summer suit and marine life-saving suit during 6-hour flight were subjects for determination of average weighted skin temperature in order to objectively evaluate human heat state and associated characteristics of operator's activities. The results of measurements aided by devices appeared more adequate, if compared to subjective pilot's estimate. PMID- 11019546 TI - [The importance of dermatological studies within sanitary measure system at industry processing natural gas with high hydrogen sulfide content]. AB - Enterprises processing natural gas with high hydrogen sulfide content appear to be alarming for prophylaxis of occupational skin diseases. Dermatologic symptoms and functional state of skin demonstrate direct and mediated unfavorable influence of industrial factors on workers and lead to prophylactic and sanative measures. PMID- 11019547 TI - [The specifics of pneumoconiosis formation under the influence of highly aluminous mullitic refractories dust (an experimental study)]. PMID- 11019548 TI - [The evaluation of combined effects of lead-copper and lead-zinc binary mixtures (experimental study)]. PMID- 11019549 TI - [Morphological characteristics of bronchial and pulmonary lavage under the exposure to brown coal dust]. PMID- 11019550 TI - [Element composition of some biological mediums in humans under chronic mercury intoxication]. PMID- 11019551 TI - Beware the prescription pitfall. Be suspicious. Be alert. PMID- 11019552 TI - "Can't you just give me some drugs to ease this pain"? PMID- 11019553 TI - They still don't get it. PMID- 11019554 TI - Tell it to the judge. PMID- 11019555 TI - Lifesaving oral cancer screening. AB - Each year 605 New York State residents, on average, die from oral and pharyngeal cancer. Disappointingly, the mortality rate has not changed significantly in more than 25 years. Early detection and risk factor prevention are the two best public health tools to reduce mortality. Dental care workers must play an increasingly important leadership role in the fight against oral cancer. A technique for an annual oral cancer screening is described in this article. PMID- 11019556 TI - Chondrosarcoma of the anterior mandible. A case report. AB - Chondrosarcomas of the maxillofacial region are extremely rare tumors, accounting for approximately 1% of chondrosarcomas of the entire body. This case represents the 12th case of mandibular symphyseal chondrosarcoma reported in the literature. It presented as a painful swelling of the alveolus, associated with a mobile mandibular incisor. The patient remains free from recurrence six years after a marginal resection. PMID- 11019557 TI - Computerizing your operatory ... and living to tell about it. PMID- 11019559 TI - Education department publishes proposal for implementing new dental anesthesia law. PMID- 11019558 TI - Ask your dentist. A campaign for dentistry. PMID- 11019560 TI - [Prevalence of problem drinkers among dental students]. AB - In 1997 and 1998 we statistically studied fourth-year dental students for their drinking and smoking habits. The subjects numbered 103 (65 males, 38 females) in 1997 and 127 (79 males, 48 females) in 1998, totaling 230. The questionnaire included the Kurihama Alcoholism Screening Test (KAST), the Adolescent Alcohol Involvement Scale (AAIS), and the Fagerstrom Tolerance Questionnaire (FTQ). Problem drinkers as defined by KAST, scoring at least 2 points, represented 12.6% of the total in 1997 and 13.4% in 1998, exceeding 10% in both years. Alcoholic misusers, with at least 42 point on the AAIS, stood at 19.4% in 1997 and 17.3% in 1998. The incidence of students assessed as having 6 points on the Quantity Frequency Scale (QF Scale) stood at 18.4% in 1997 and 10.2% in 1998. Compared between two sexes, KAST detected no significant difference between the two groups: 13.9% for males and 11.6% for females. Alcoholic abusers on the AAIS, however, were estimated at 22.9% in males and 9.3% in females, the ratio for males being more than twice as high as that for females. It was also found that 47.9% of the male students and 33.7% of their female counterparts drank every weekend and that 31.3% of the males and 5.8% of the females kept drinking until they were drunk. Female students thus appeared to drink more frequently, though they consumed smaller amounts of alcohol at a time. The smoking rate was lower than the average in the Japanese for both groups; nicotine dependence was also lower in dental students. It was demonstrated that dental students drink more alcohol more frequently, with a higher incidence of problem drinkers. PMID- 11019561 TI - [A study of recovery factor about drug addiction recovery center "DARC"]. AB - This study examines the effectiveness of the 'DARC' (Drug Addiction Rehabilitation Center) and its program in treating patients with drug dependence. DARC is the only rehabilitation facility group managed by recovered drug users in Japan. It uses the 'Twelve Step Program' as part of its recovery regime. Through structured questionnaires, the changes of participants' and ex-participants' everyday behavior, their acceptance of spirituality, attitude toward drugs, self esteem, their coping skills and loneliness were examined. The results revealed: a.. Negative partial correlation between loneliness and everyday behavior. b.. Negative partial correlation between loneliness and positive/effective coping skills. c.. Positive partial correlation between acceptance of 'spirituality' and attitude toward drugs. d.. Positive partial correlation between acceptance of 'spirituality' and positive/effective coping skills. These results suggest that the acceptance of 'spirituality' and decreasing loneliness are important factors in the recovery from drug dependence. PMID- 11019562 TI - [Outcomes and predictors of mortality in elderly patients requiring artificial ventilation]. AB - We retrospectively examined the outcomes and the predictors of mortality in 97 patients aged 70 years and over (mean: 79.3 years) who required artificial ventilation for more than 3 hours. The median duration of artificial ventilation was 16 days (range: 1-85). Of these patients, 61% survived ventilator weaning and 37% were discharged from hospital alive. We performed univariate and logistic regression analysis to determine the predictors of dying before weaning and hospital discharge using severity of illness data. The predictors of hospital mortality were examined in 86 patients, excluding those who had malignant disease, all of whom died in hospital. Activities of daily living (ADL) were ranked as "bedridden", "in wheelchair", or "independent". In the three age groups up to 70 years, 75 to 84 years and 85 years and over-the respective survival rates were 63% (weaned) and 67% (discharged), 69% (weaned) and 39% (discharged), and 33% (weaned) and 12% (discharged); the overall p values being 0.026 (weaned) and 0.003 (discharged). The predictors of dying before weaning according to univariate analysis were as follows: age (p = 0.026), respiratory or cardiac arrest on admission (p = 0.003), acute physiology score (APS) of 25 or more on admission (p = 0.000), systolic blood pressure below 90 mmHg on admission (p = 0.001), hemoglobin less than 11 g/dl (p = 0.044), and total protein less than 6 g/dl (p = 0.007). The predictors of hospital mortality by univariate analysis were as follows: age (p = 0.003), limited ADL (p = 0.001), respiratory or cardiac arrest on admission (p = 0.011), APS 25 or more on admission (p = 0.049), systolic blood pressure less than 90 mmHg on admission (p = 0.002), hemoglobin less than 11 g/dl (p = 0.028), and GOT or GPT more than 50 IU (p = 0.038). The relative risk of dying before weaning decreased in the order: respiratory or cardiac arrest on admission, systolic blood pressure less than 90 mmHg on admission, total protein less than 6 g/dl (Odds ratios: 6.04, 3.90, 3.51, respectively), or, respiratory or cardiac arrest on admission, APS more than 25 in admission, total protein less than 6 g/dl (Odds ratio: 6.94, 3.99, 3.76, respectively). The relative risk of hospital mortality decreased in the order: "bedridden", systolic blood pressure less than 90 mmHg on admission, "with wheel chair" (Odds ratios: 11.76, 6.44, 3.57, respectively). In the older patients, successful ventilator weaning was not indicative of hospital discharge. Ventilator weaning depended mainly on acute health status on admission, but hospital discharge depended also on the presence of limited ADL and preexisting malignant disease. PMID- 11019563 TI - [Evaluation of population-based lung cancer screening in Niigata and analysis of interval cases based on comparison lung cancer registry with screening records]. AB - Survival analysis has been used as one way to examine the efficacy of cancer screening. However, this type of study is susceptible to many sources of bias, especially to a length bias. In the city of Sukumo in Japan, a survival analysis considering screening history for the purpose of eliminating the length bias was conducted. The efficacy of lung cancer screening was assessed by comparing those who participated in the screening in the year preceding diagnosis, with those who did not. The 5-year survival rate was 44% in the former and 16% in the latter. We tried to evaluate population-based lung-cancer screening in Niigata using the method as that of Sukumo by comparing lung-cancer registry data with screening records, those who died of lung cancer from 1991 to 1994 in the study area and had the opportunity to be screened were categorized according to the screening history for the year preceding diagnosis. The 5-year survival rate was 41% in those who were screened and 19% in those not screened. In addition, the median survival time of the former was 37 months, significantly longer than the 12 months of the latter. Replication of study results in different populations may suggest the efficacy of lung-cancer screening on an annual basis. We also analyzed the clinical background of 47 interval cases in the search for a future direction for the improvement of the screening method. PMID- 11019564 TI - [Mortality and prognostic factors in patients with community-acquired pneumonia: an analysis of 231 cases]. AB - We prospectively analyzed the mortality and prognostic factors in 231 patients with community-acquired pneumonia. The patients were enrolled between May 1996 and April 1998. The average patient age was 67.6 +/- 17.5 years, and 68% were male. The mean hospital length of stay was 21.9 +/- 21.7 days. The mortality was 6.5% at the 30th day, 13.9% at hospital discharge, and 19.9% at 1 year later. Stepwise logistic regression analysis showed that liver cirrhosis, diastolic hypotension (< 60 mmHg), hypoxemia (< 50 Torr) were significantly correlated with death at the 30th day, and that alcoholism, malignancy, diastolic hypotension, hypoxemia, hypoalbuminemia (< 3.0 g/dl), and increased creatinine (> 1.2 mg/dl) were significantly correlated with discharge mortality. Furthermore, the prognostic factors at 1 year later were the same as those at hospital discharge, with the exception of alcoholism, which was replaced by ischemic heart disease in the 1-year mortality. We conclude that liver cirrhosis, diastolic hypotension, and hypoxemia are very important prognostic factors during the acute stage of pneumonia, and that alcoholism, malignancy, hypoalbuminemia and renal complications are significantly correlated with hospital death and 1-year mortality. PMID- 11019565 TI - [A case of primary macroglobulinemia with acute pulmonary edema]. AB - A 54-year-old man with acute respiratory failure underwent emergency admission to our hospital. Acute pulmonary edema combined with severe pneumonia was strongly suspected, and treatment with diuretics, antibiotics, and corticosteroids was effective. The diagnosis, primary macroglobulinemia, was made from the following findings: a high level of the lgM 1 type monoclonal protein, and an elevated serum viscosity with characteristic "linked sausage effects" seen in the retinal veins. Lymphoplasmacytoid cells, plasma cells, and small lymphocytes were seen in the bone marrow and lymphoplasmacytoid cells were also seen in the smear of the patient's peripheral blood. The types of malignant cells found in his bone marrow were also seen in both infiltrative lesions in the lung parenchyma and the pleural effusion. We concluded that this was a case of primary macroglobulinemia with acute pulmonary edema, which was caused partly by the hyperviscosity syndrome. The antitumor effect of the corticosteroid might have some effect on the recovery from acute respiratory failure and primary macroglobulinemia. He was transferred to the Department of Hematology to receive suitable chemotherapy. PMID- 11019566 TI - [Relapse of idiopathic chylothorax 17 years after remission of congenital chylothorax]. AB - We report a case of chylothorax in a 17-year-old male. As a neonate, had had congenital chylothorax, and was successfully treated with medium-chain triglycerides; but recently presented with dyspnea after an episode of severe coughing. Radiographic examination disclosed abundant effusion in the right chest. The effusion was milky brown, had a creamy supernatant and a high triglyceride level (3085 mg/dl). This condition was diagnosed as idiopathic chylothorax. Lymphangiography showed a rupture of the right thoracic duct. Since the effusion was resistant to conservative therapy, we performed thoracoscopic clipping of the thoracic duct, which reduced the amount of pleural effusion. We speculated that coughing may have caused the rupture of a congenitally weakened thoracic duct. PMID- 11019567 TI - [A case of adenocarcinoma complicated with massive leukocytosis and hypercalcemia]. AB - A 66-year-old man was admitted to our hospital complaining of non-productive cough and low-grade fever. Chest X-ray examination revealed a mass shadow in the right hilum. Transbronchial lung biopsy of the tumor mass yielded a diagnosis of adenocarcinoma. Despite repeated chemotherapy using CDDP and VDS, metastasis to the right adrenal gland and right femur occurred, and was accompanied by hypercalcemia and hypophosphatemia. Serological study revealed elevated levels of PTH-rP and G-CSF. Six months after adenocarcinoma was diagnosed, multiple skin metastases of the cancer were observed. Immunohistochemical staining for PTH-rP and G-CSF indicated that production of cytokines had caused a paraneoplastic syndrome including hypercalcemia and leukocytosis. It appeared that the elevation of G-CSF was induced by IL-6 produced from PTH-rP in cancer tissue. Documentation of similar cases is required. PMID- 11019568 TI - [A case of hypersensitivity pneumonitis caused by Strophariaceae]. AB - A 38-year-old woman was admitted to the Iiyama Red Cross Hospital with a non productive cough. She had been working with Strophariaceae cultures for 4 years, and in January 1999 a cough had developed. On admission, hypoxemia was noted. The serum level of lactate dehydrogenase was slightly elevated. Chest radiography and CT scanning showed diffuse ground glass opacity and small centrilobular nodules in both lower lobes. In the bronchoalveolar lavage fluid, the total cell count was normal, the CD4/CD8 ratio was reduced, and the lymphocyte fraction was very high. A transbronchial lung biopsy revealed alveolitis with lymphocytic infiltration and organizing changes like Masson bodies. The lymphocyte stimulation test was positive for Strophariaceae. Hypersensitivity pneumonitis caused by Strophariaceae was diagnosed. Prednisolone was administered because of persistent symptoms and prolonged abnormal chest radiography findings. These abnormalities disappeared 2 weeks after prednisolone treatment was initiated. There is one previous report of hypersensitivity pneumonitis caused by Strophariaceae. The symptoms are coughing and sputum production as in the common cold. The occupational history must be carefully considered in making a diagnosis. PMID- 11019569 TI - [Serrapeptase-induced lung injury manifesting as acute eosiniphilic pneumonia]. AB - An 84-year-old man was referred to our hospital because of fever, cough, and hemoptysis. The patient had acute respiratory failure (PaO2 < 40 mmHg) on admission, with diffuse interstitial infiltration and bilateral pleural effusion. The bronchoalveolar lavage fluid was bloody, and contained a high percentage of eosinophils (32%). A diagnosis of acute eosinophilic pneumonia was established, and the patient made a rapid recovery after corticosteroids were administered. When the DLST (drug lymphocyte stimulation test) was performed after the corticosteroid therapy was stopped, it was positive for serrapeptase, which had been prescribed for chronic cystitis for 3 months before the onset of the pneumonia. This was a case of drug (serrapeptase)-induced pneumonitis manifesting as acute eosinophilic pneumonia. PMID- 11019570 TI - [A case of idiopathic subglottic stenosis successfully treated with endoscopic laser treatment]. AB - A 24-year-old woman presented with an abnormal sound in her neck. She had no history of previous surgery, endotracheal intubation or neck trauma. She had been aware of this abnormal sound since she was 12 years old, but paid no attention to it. She had been recommended to visit a hospital for a neck examination. Stridor was heared in the neck ["coming from the throat"?]. Peak flow was markedly suppressed in pulmonary function tests, and the flow volume loop indicated upper airway stenosis. Bronchoscopy revealed subglottic tracheal stenosis, and magnetic resonance imaging showed that the cricoid cartilage was normal. Nd-YAG laser treatment was performed in Mitsui Memorial Hospital and her symptoms were improved. Biopsy of the stenotic area of the trachea showed dense fibrous tissue proliferation in the submucosa. Neither granuloma nor vasculitis was found in the biopsy specimen. No systemic disease capable of inducing tracheal stenosis was found on systemic examination. From these clinical and pathological findings, we diagnosed idiopathic subglottic stenosis. PMID- 11019571 TI - [Occurrence of BOOP outside radiation field after tangential radiation therapy for breast carcinoma]. AB - We report three cases of bronchiolitis obliterans organizing pneumonia (BOOP) that occurred outside the radiation field after radiation therapy using tangential fields for breast carcinoma. All patients complained of a cough between 14 and 20 weeks after completion of radiation therapy. Fever also developed in two of the three. Chest radiography and computed tomography demonstrated peripheral alveolar opacities outside the radiation field on the same side as the radiation therapy. Laboratory data showed an increased level of C-reactive protein and an increased erythrocyte sedimentation rate. Bronchoalveolar lavage showed an elevated total cell count with a very high percentage of lymphocytes. Transbronchial lung biopsy revealed a histologic pattern consistent with BOOP. Treatment with corticosteroids resulted in rapid clinical improvement and complete resolution of the radiographic abnormalities. This pulmonary disorder appears to be induced by radiation, especially when a tangential field is employed for breast carcinoma, though the etiology has not been fully investigated. It is important to be aware of this type of pulmonary complication in patients given radiotherapy for breast carcinoma. PMID- 11019572 TI - [A case of Klebsiella pneumoniae infection causing a buccal abscess complicated with multiple lung abscesses]. AB - A 51 year-old man fitted with a dental prosthesis was hospitalized with buccal swelling, fever and chest pain. Laboratory data showed marked inflammatory changes, and chest radiography and CT scanning revealed small nodular shadows within the lung. A diagnosis of multiple lung abscesses secondary to a buccal abscess possibly caused by the prosthesis was made from needle aspiration biopsies of the lung nodules and of a buccal lesion. Klebsiella pneumoniae was isolated from these lesions and from a blood culture. The patient was successfully treated with antibiotics and by surgical drainage of the buccal abscess. It is important to note that the patient was immunodeficient at the time as a result of diabetes and alcohol intoxication. PMID- 11019573 TI - [A case of Wegener's granulomatosis with pachymeningitis]. AB - A 62-year-old woman who had been receiving corticosteroid therapy for pachymeningitis since 1997 was admitted to our hospital when an abnormal shadow was noticed in her chest radiograph. In bronchial and nasal mucosal biopsies, the findings of a necrotic granulomatous lesion and vasculitis were compatible with Wegener's granulomatosis, although this is rarely seen with pachymeningitis. After further corticosteroid therapy together with cyclophosphamide treatment, the size of the thoracic X-ray shadow decreased. Methicillin-resistant Staphylococcus aureus (MRSA) was cultured from the sputum and the nasal fluid, and may have contributed to the advance of the disease in the airway. This case will require continuing careful observation. PMID- 11019574 TI - [A case of isolated sarcoidosis of the cerebral falx]. AB - A 28 year-old man complaining of intermittent headaches for a month following an episode of general convulsions was admitted to the Department of Neurosurgery of our hospital. His brain CT scan showed a well-enhanced mass growing symmetrically along the anterior cerebral falx in a manner compatible with meningioma. Nothing abnormal was detected in chest radiography on admission. Histopathological examination of the mass after partial resection revealed multiple epithelioid cell granulomas with giant cells indicating sarcoidosis. After 4 weeks of treatment with oral prednisolone 30 mg/day, bronchoalveolar lavage was performed in our department. In the lavage fluid, both the total cell count and the percentage of lymphocytes were elevated. The lung specimens obtained by transbronchial biopsy also showed epithelioid cell granulomas. CNS sarcoidosis is reportedly usually observed in the vicinity of the hypothalamus with other organ involvement. In the present case, differential diagnosis of sarcoidosis from meningioma was difficult because the solitary lesion was growing along the anterior cerebral falx without radiographic evidence of bilateral hilar lymphadenopathy. PMID- 11019575 TI - [A case of tracheobronchomegaly]. AB - We report a rare case of tracheobronchomegaly with crescent-type tracheobronchomalacia. A 77-year-old man with a chronic cough was referred to our hospital because of fever and dyspnea. Radiographic examination showed enlargement of the trachea and main bronchi. On chest radiography, the transverse diameter of the trachea was 31 mm, and consolidation shadows were seen in both upper lung fields. Tracheobronchomegaly with pneumonia was diagnosed. The pneumonia was improved by administration of PAPM/BP. On bronchoscopic examination, the trachea and main bronchi were extremely dilated on inspiration, and were collapsed on expiration. The biopsy specimen from the bronchial mucosa showed non-specific chronic inflammation. PMID- 11019576 TI - [Contrast-enhanced three-dimensional angiography]. AB - Rapid advances in the techniques of contrast-enhanced MR angiography (MRA) have made it possible to evaluate the entire aorta and main arteries as well as small arteries and veins of the whole body. In this article, the technical aspects and clinical applications of contrast-enhanced MRA are reviewed. Dynamic contrast enhanced three-dimensional (3D) MRA is a first-pass vascular imaging technique that uses the 3D fast field echo sequence with rapid bolus injection of contrast material. By repeating imaging sequences, it is possible to trace the dynamics of the injected contrast material over time and apply various postprocessing techniques, such as double subtraction and addition, to demonstrate arteries and veins separately and to display the arteries in a single image, even when circulation time is prolonged. This technique is flexible enough to be applied in a variety of clinical settings by optimizing the examination. The abdominal aorta and main arteries of the lower extremities are examined by two methods, dynamic subtraction 3D MRA (MRDSA) and the moving-bed method. Three bolus injections of contrast material are used in MRDSA. In contrast, a slow continuous injection of a large dose (20 ml or more) of contrast material is used in the moving-bed method, which is a convenient, useful method of contrast-enhanced MR angiography when the study must focus on arterial imaging. Clinical indications include atherosclerotic occlusive disease, Takayasu arteritis, aortic dissection, anti phospholipid antibody syndrome, aneurysm, patent and occluded bypass graft, and deep vein thrombosis, among others. The problem with contrast-enhanced MRA is its difficulty in depicting small vessels such as mesenteric arteries, distal renal arteries, and peripheral small arteries. The nature of gradient-echo sequences has certain inherent problems for the imaging of small vessels. The fat-tissue interface becomes dark because of chemical shift artifact (which occurs both when the echo time is set in-phase and when it is set out-of-phase), reducing the ability to detect small arteries with contrast-enhanced MR angiography. The fat suppression technique is very useful for satisfactorily demonstrating small vessels by reducing chemical shift artifact. MR angiography remains an exciting and challenging field, and the best methods of contrast-enhanced MR angiography will be developed and established on the foundation of all previous work. PMID- 11019577 TI - [Clinical application of endoscopic ultrasonography (EUS) for gastrointestinal diseases]. AB - Endoscopic mucosal resection (EMR) has been widely employed as a minimally invasive surgical procedure for early digestive cancers. Moreover, many recent reports emphasize the usefulness of endoscopic ultrasonography (EUS) for diagnosis of early gastrointestinal cancers. Generally, the normal gastrointestinal wall is visualized as having five layers by endosonographic probes of 7.5, 12, or 20 MHz. Delineation of these layers is the most important point for feature of EUS. Only mucosal cancers that are disclosed as hypoechoic masses within the first to second layer are indicated for EMR. The development of further extracorporeal applications such as color Doppler, 3-D, and aspiration biopsy EUS will contribute to the increased use of EUS in the near future. PMID- 11019578 TI - [Advanced gastric cancer: the findings of delayed phase dynamic CT and radiologic histopathologic correlation]. AB - PURPOSE: The aim of this study was to describe delayed phase dynamic CT findings of advanced (T2-T4) gastric cancer and to correlate with histopathologic findings. METHOD/MATERIALS: Quadruple phase dynamic CT including delayed imaging taken five minutes after the start of injection of contrast material was performed in 43 patients with 45 advanced gastric cancer and 20 control subjects with no gastric lesions. On delayed phase CT scans, the attenuation of the gastric wall was equal to or lower than that of the liver parenchyma in the control subjects, therefore, the presence of higher attenuation in the gastric wall was considered to be abnormal and defined as delayed enhancement. Histopathologic findings in the tumors showing delayed enhancement were compared with those in the tumors without this feature. RESULTS: Delayed enhancement was seen in 26 (57%) of the 45 tumors. Eleven of 25 differentiated-type tumors and 15 of 20 undifferentiated-type tumors showed delayed enhancement (P < .05). Delayed enhancement was seen in one of five medullary type tumors, in 11 of 25 intermediate-type tumors, and in 14 of 15 scirrhous-type tumors (P < .005). CONCLUSIONS: Delayed enhancement was frequently seen in the tumors with abundant fibrous tissue stroma. Delayed phase dynamic CT may be useful for the characterization of advanced gastric cancer. PMID- 11019579 TI - [Gradually enhancing breast cancer on dynamic MRI]. AB - PURPOSE: The aim of our study was to evaluate the frequency and the radiologic and pathologic features of gradually enhancing breast cancers on contrast enhanced dynamic MRI. MATERIAL AND METHODS: 83 patients with pathologically proven breast cancer underwent contrast enhanced dynamic MRI. Breast cancers that showed a gradually enhancing pattern were selected, and their pathological features and the findings in other imaging modalities were retrospectively analyzed. RESULTS: Of 83 lesions, 8 lesions (10%) showed a gradually enhancing pattern on dynamic MRI. The lesions included 3 papillotubular carcinomas, 3 scirrhous carcinomas, one solid-tubular carcinoma, and one mucinous carcinoma. All of them had imaging features suggesting malignancy on mammography and ultrasonography. Histopathologically, all 7 invasive ductal carcinomas showed prominent fibrosis in the stroma, and one showed many dilated ducts. One mucinous carcinoma showed large mucinous pools. CONCLUSION: Some breast cancers may show a gradually enhancing pattern on dynamic MRI. Other imaging features should be taken into account in the differential diagnosis of breast tumors. PMID- 11019580 TI - [Comparison of detection pattern of HCC by ferumoxide-enhanced MRI and intratumoral blood flow pattern]. AB - We compared the detection rate and pattern of ferumoxide-enhanced magnetic resonance imaging (Fe-MRI) with the intratumoral blood flow pattern determined by CT angiography (CTA) and CT portography (CTAP) in 124 nodes (34 cases) diagnosed as hepatocellular carcinoma (HCC) or borderline HCC, based on the clinical course. Sequences to obtain a T1-weighted images (T1W), proton density-weighted images (PDW), T2-weighted images (T2W), T2*-weighted images (T2*W) were used in Fe-MRI. In nodes shown to be hypervascular on CTA, the detection rate by Fe-MRI was 69.7%. In nodes shown to be avascular by CTAP, the detection rate by Fe-MRI was 67.3%. These rates were higher than with other flow patterns. In nodes showing high signal intensity (HSI) on any sequences, arterial blood flow was increased and portal blood flow decreased in comparison with nodes without high signal intensity. All nodes showing HSI, both on Fe-MRI T2W and T2*W, were hypervascular on CTA, and portal blood flow was absent on CTAP. Nodes showing HSI on both T2*W and T2W were considered to have greater arterial blood flow and decreased portal blood flow compared with nodes appearing as HSI on T2*W, but only as iso- or low signal intensity on T2W (Mann-Whitney U-test; p < 0.05). PMID- 11019581 TI - [MR angiography of hemodialysis access fistula: evaluation before PTA procedure]. AB - We evaluated the feasibility of MR angiography for hemodialysis fistula. Eleven patients with suspected stenosis or occlusion of an autogenous hemodialysis fistula in the forearm underwent MRA. MRA was obtained using both the 3D-PC method and 3D gadolinium contrast-enhanced (CE) method with a knee coil. The two methods were compared with DSA in seven patients. CE-MRA clearly demonstrated the entire dialysis fistula. However, signals of the stenotic area are effaced in PC MRA. The diameter of the vessels evaluated in the CE-MRA was well correlated with angiography. CE-MRA demonstrated smaller collateral vessels more clearly than PC MRA. CE-MRA provided useful information before the PTA procedure. PMID- 11019582 TI - [Auditory cortical response to monaural stimulation as detected by functional magnetic resonance imaging]. AB - In order to confirm the crossed-innervation between auditory cortex and the ear that receives monosyllabic sound, the auditory cortical response to monaural monosyllabic stimulation as detected by functional magnetic resonance imaging (fMRI) was investigated in six normal hearing subjects. Stimulus amplitude averaged 95 dBSPL at the distal end of the audio system. A series of 440 echo planar images was acquired during the acoustic stimulation within the four OFF-ON cycle paradigm. Five image series with 10 slices were collected within each OFF or On period. Each scanning session began with four baseline images before the OFF-ON paradigm. Monosyllabic sounds were presented monaurally during the ON period at a rate of one monosyllable/sec. Functional MRI data were analyzed with SPM99b software (Statistical Parametric Mapping). The background scanner noise averaged 97dBSPL. The selicon ear plug and headphone as acoustic shields attenuated the noise as much as 17 dB. A broad and intense auditory cortical response was observed bilaterally in response to monaural monosyllable stimulation. Sound presentation to the right ear was followed by a larger response in the left auditory cortex than in the right, and left ear stimulation evoked a larger response in the right auditory cortex than in the left. This pattern was consistent in all subjects examined. The primary auditory cortex responded to monosyllabic words presented to the contralateral ear. The results confirmed the crossed-innervation between the auditory cortex and ear for listening to monosyllables. Functional MRI is a useful tool for investigating auditory cortex function, if the scanner noise is adequated controlled. PMID- 11019583 TI - [Automated infant auditory screening using the Natus-ALGO 2e in the NICU]. AB - Natus-ALGO 2e, an automated ABR screener(Natus Medical, Foster City, CA, USA), compares the V wave of ABR evoked by 35-dB-nHL click stimuli by using a template matching detection algorithm that provides only a pass-refer outcome. The aim of this study was to compare Natus-ALGO 2e with conventional ABR, and to evaluate its usefulness. The Natus-ALGO 2e screener was used to screen 202 ears of 101 neonates in our neonatal intensive care unit. The mean conceptional age at the time of screening was 40.4 +/- 3.0 weeks. 60 ears of 30 infants at high-risk of hearing impairment, including "refer" infants, were tested by the Natus-ALGO 2e and conventional ABR methods, and the results were compared. All neonates were tested with the Natus-ALGO 2e screener in a state of natural sleep, and screening time averaged 2 minutes 58 seconds. There were 97 cases in which both ears were passed, 3 cases in which both ears were referred, and one case in which one ear was referred. In comparison with conventional ABR, 53 of the 60 ears of 30 high risk infants passed by the Natus-ALGO 2e method, whereas 14 of the 53 ears initially failed the conventional ABR screening. Of these 14 ears disagreements (the results of the Natus-ALGO 2e method passed, but the results of the conventional ABR failed), the results of the ABR screening changed to normal in 11 ears, and ABR showed improved threshold and latency in the other 3 ears after 5 weeks to 12 months. Among those that passed the Natus-ALGO 2e screening, the number of sweeps that failed the ABR screen was significantly greater than with normal ABR. Of the 7 ears of 4 patients that were referred on the basis of the Natus-ALGO 2e screening and failed by the conventional ABR method, 3 ears screened by the ABR method were normal when retested, and one ear passed by the Natus-ALGO 2e screening 12 weeks to 11 months later. In conclusion, Natus-ALGO 2e is useful for screening infant hearing because it can be performed quickly while the patient is sleeping naturally. In infants at high-risk for hearing impairment, the results of Natus-ALGO 2e and conventional ABR screening conflicted in numerous sweeps. Therefore, when there are many sweeps in high-risk infants, a retest should be performed that includes conventional ABR, even if they passed with Natus-ALGO 2e. PMID- 11019584 TI - [Concurrent chemoradiotherapy for locally unresectable head and neck cancer]. AB - In patients with locally unresectable head and neck cancer with large nodal involvement, the expected five-year survival is as low as 1-2%. To improve the prognosis of these patients, we studied the usefulness of concurrent chemoradiotherapy in a phase II trial. Between September 1996 and May 1999, thirty-five patients with locally unresectable head and neck cancer were administered concurrent chemoradiotherapy consisting of low-dose and long-term treatment with cisplatin (CDDP) plus 5-fluorouracil (5FU), or (L-CF); the L-CF regimen consisted of CDDP, 3 mg/m2 on 5 days of the week and 5FU, 150 mg/m2 as a 24-hour infusion on 5 days of the week. Concurrently, conventional radiotherapy was given up to total dose of around 60 Gy. In the 33 patients evaluable for response, 17 complete and 9 partial responses were noted, with an overall response rate of 79%. Oral mucostis and myelosuppression were the major side effects and mucositis was a dose limiting toxicity. This study demonstrates increase in survival among the responders (complete + partial) in the concurrent chemoradiotherapy setting. However 8 local relapses were eventually noted in the 17 complete responders. We concluded that this treatment strategy was beneficial in patients with locally unresectable head and neck cancer. PMID- 11019585 TI - [Imaging studies and surgical approach to postoperative maxillary cysts]. AB - Surgical treatment for postoperative maxillary cysts (POMC) can be performed using the peroral method, or the endonasal method which is less invasive. Recently, the percentage of cases in which the endonasal method is employed has increased with the widespread use of endoscopy and advancement in imaging techniques. However, the peroral method is still necessary in some cases. During the 5-year period from July 1994 to June 1999, we operated on 29 patients (31 sides) for POMC. The endonasal method was used for 22 sides, and the peroral method was needed for the remaining 9 sides. Following analysis of the CT scans and MR images, we concluded that it is difficult to employ the endonasal method in the following cases; 1) Cysts located in the lateral or anteroinferior portion of the maxillary sinus 2) Small cysts 3) Cysts with a bony wall. PMID- 11019586 TI - [Usefulness of electroglottogram (EGG) and photoglottogram (PGG) for the analysis or vocal fold vibration--a high speed digital imaging study]. AB - Simultaneous recordings of EGG, PGG and high-speed digital imaging (HSDI) of vocal fold vibration were made in 3 normal male subjects and the results were compared to evaluate the usefulness of EGG and PGG for the analysis of the vibratory pattern. Each subject was required to produce sounds of 3 different pitches at 3 different intensity levels in the recording sessions and the 27 sets of data samples were collected. For each data set, the glottal area waveform (GAW), glottal width waveform (GWW), EGG, differentiated EGG, PGG, differentiated PGG, and voice waveform were displayed in a graphic form on the same time axis and their patterns were compared. Based on the analysis of GAW and GWW, it was found that there were variations in the pattern of closing and opening of the vocal fold margin corresponding to the differences in the type of phonation. The EGG waveform appeared to correspond well to that of the movement of the vocal fold margin in the closing phase, while the correspondency was less apparent in the opening phase. It is considered that the EGG waveform is dependent on the mode of contact between the two vocal folds and that a careful observation of the EGG is useful and practical for predicting the pattern of vocal fold vibration. On the other hand, it was difficult to determine the onset of the closing and opening phases only from PGG waveform and there was a timing difference between the PGG and GAW. Although PGG seemed to be useful for judging the presence or absence of the opening phase and for the pitch measurement, it is considered to be unsuitable for a precise cycle-by-cycle analysis of the pattern of vocal fold vibration. PMID- 11019587 TI - [Ocular counter-rolling after prolonged alteration in the direction of gravity]. AB - To investigate the dynamics of otolith and oculomotor function, we subjected volunteers to a lateral body tilt for a period of two hours and analyzed ocular counter-rolling. Six healthy adult volunteers were roll-tilted along the naso occipital axis at a speed of 0 degree/sec. from the 0 degree earth vertical position to the 90 degrees lateral tilt position. After two hours, the volunteers were returned to the 0 degree earth vertical position. Ocular counter-rolling was recorded using an infrared CCD camera. The video recordings were made in the dark while the volunteers had their eyes open. Recordings were obtained (1) as the volunteers was moved from the 0 degree earth vertical position to the 90 degrees lateral position, (2) 5 minutes after reaching the lateral position, (3) during the roll-back from the 90 degrees lateral position to the 0 degree upright position, two hours after the lateral tilt loading, and (4) 5 minutes after reaching the final upright position. The occurrence of ocular counter-rolling between the 0 degree upright position and the 30 degrees tilt position was confirmed during both roll movements from 0 degree to 90 degrees and from 90 degrees to 0 degree. The counter-rolling was most noticeable between the 0 degree and the 30 degrees positions. No differences in ocular counter-rolling during the roll-tilt and the roll-back situations were observed. These results indicate that the two-hour lateral tilt position did not produce any functional changes in the hair cells and/or the otolith-oculomotor system. All of the subjects exhibited ocular counter-counter-rolling during the initial stage of their roll-back to a normal upright position. This phenomenon might result from the additional bending of the hairs by inertia during the initiation of the backward roll motion. PMID- 11019588 TI - [Assessment of the psychological stress experienced before and after the use of a hearing aid in elderly patients]. AB - Hearing aids are useful instruments for elderly patients with impaired hearing. However, the psychological effects of wearing a hearing aid have remained unclear until recently. A study was therefore performed at the Tokyo Metropolitan Geriatric Hospital to determine such effects. Eighteen out of 50 questions included in the "Psychological Stress Response Scale (PSRS)" were selected and used to produce a questionnaire. The questionnaire was then given to and completed by 31 elderly patients who received and willingly used hearing aids. Of these patients, 20 were female and 11 were male. Their average age was 80.4 years (+/- 5.3 years ranging from 66-89 years). The questionnaire was completed by each patient on two separate occasions: before the hearing aids were fitted and after. In addition to the PSRS questionnaires, a second "hearing satisfaction" questionnaire was also completed, both before and after the hearing aids were fitted. The scores for depression, anxiety and anger were significantly lower after the hearing aid had been fitted. The score for depression showed a particularly distinctive decrease. The statistics also showed that the use of a hearing aid produced a significant improvement in the level of satisfaction patient's felt regarding their hearing. We conclude that the use of hearing aids by elderly patients not only alleviates their dissatisfaction with being unable to hear, but has additional psychological advantages. These results suggest that the use of a hearing aid not only improves hearing but may also delay or offset the development of dementia. Accordingly, suitable hearing aids should be provided to elderly patients so that they may recover their hearing and benefit from the associated psychological advantages. PMID- 11019589 TI - [Detection of varicella-zoster virus DNA in tear fluid and saliva of patients with Ramsay Hunt syndrome]. AB - Ramsay Hunt syndrome develops when the varicella-zoster virus (VZV) is reactivated. In the present study, we examined the secretion kinetics of VZV DNA in the tear fluid, submandibular gland saliva and parotid gland saliva of 15 patients with Ramsay Hunt syndrome. The presence of VZV DNA was detected using PCR and a microplate hybridization method. Hybridization signals were measured using the fluorescence density of an enzymatic reaction product using fluoroscan and a system involving streptavidin-conjugated beta-galactosidase. The results were converted into numerical values and used to estimate the number of virus DNA copies. VZV DNA was detected in the tear fluid, submandibular gland saliva and parotid gland saliva of the Ramsay Hunt syndrome patients. The rate of VZV DNA detection in the submandibular gland saliva was 72%, and the detection rate in the parotid gland saliva was 57%. The detection rate in the tear fluid was 27%, which is significantly lower than other two detection rates. Regarding the submandibular gland saliva and the parotid gland saliva, the VZV DNA was detected in samples collected at a comparatively early stage of onset. In the tear fluid, the detection rate increased significantly in samples collected 2 weeks after onset or later. Thus, differences in the detection rate were observed depending on the type of secretory gland and the timing of the sample collection. The VZV DNA in the tear fluid is thought to derive from the ganglion trigeminale. The increase and decrease in the number of VZV DNA copies detected in samples collected at different times is considered to substantiate VZV reactivation in Ramsay Hunt syndrome. PMID- 11019590 TI - [Influence of vestibular input on visually guided saccades]. AB - A common pathway in the brainstem is thought to exist for the induction of visually guided saccades and rapid phase nystagmus (Ref. Ohki et al. 1988). This common pathway is often referred to as "the saccade generator" or "the burst generator." The presence of this pathway suggests that vestibular function can be examined by applying "visual stimuli" and "vestibular stimuli" at the same time. We therefore investigated whether caloric stimulation affects the latency, amplitude and velocity of saccadic eye movements and if so, how. Nine normal volunteers participated in the experiment. Eye movements were recorded using DC electrooculography. Caloric stimulation was applied to the right ear using an air caloric stimulator (10 degrees C) during saccades. Two experiments involving visually guided saccades were performed. Task 1: The subject was asked to fixate on a central fixation point. Another spot of light then randomly appeared at one of two different locations (20 degrees to the right or left). The subject was then asked to scan towards the second light as quickly as possible. Task 2: The procedure was the same as in Task 1, except that the target appeared only briefly and then later reappeared at the same location. The saccade latency was unaffected by caloric stimulation in Task 1, while the latency of the rightward saccades increased significantly in Task 2. The saccade amplitude and velocity increased significantly in both directions in Task 1, while the amplitude and velocity of the leftward saccades increased significantly in Task 2. These results can be explained by the synergistic activation of Burster-Driving Neurons by the saccade signal and the vestibular signals. The combinations of these saccade tasks and caloric stimulation, especially Task 2, may be useful in clinically examining vestibulo-oculomotor functions. PMID- 11019591 TI - [Coronary angioplasty over 70 years of age]. AB - In the two institutions between January 1992 and December 1998 PTCA-s were done in 196 pts (123 male and 73 female) aged 70 years or more, with the 57 (20.1%) 6 months redo procedures performed out to June 1999, totally 247 interventions. The dilatations were done in 230 vessels (218 natives and 12 grafts) and in 254 stenoses, as of 1996 in 73 cases with stent deployment, too. PTCA was clinically successful in 178 (90.8%) pts, suboptimal result and/or MACE (AMI, emergency PTCA or CABG, fatal outcome) occurred in 18 (9.2%) cases. Due to unsuccessful PTCA-s, 3 (1.5%) pts underwent acute, 3 elective bypass surgery, in 1 (0.5%) case thoracal sympathectomy without revascularisation, in 1 no further intervention was done. The in-hospital as well as the 30 days mortality was 5 (2.6%). Taking into consideration the higher risk of CABG surgery at an old age, in the case of indication for coronary revascularisation the possibility of angioplasty should be considered first. PMID- 11019592 TI - [Kinetics of aluminum absorption and serum concentration in chronic renal insufficiency]. AB - After administration of a single dose of an aluminium containing antacid (Tisacid) to eight non dialysis patients in various stages of chronic renal insufficiency, the kinetics of serum aluminium absorption was studied. Serum aluminium peaked at two points of time, aluminium absorption being biphasic in renal patients comparing with monophasic kinetic in controls. At the same time, already at 24 hrs after Tisacid ingestion, the aluminium concentration was almost twice as high as initially even in mild renal insufficiency. These results are at variance with the rapidly reversible, monophasic aluminium absorption observed earlier in healthy subjects and in non-renal patients with duodenal ulcer. To ward off complications, a strong warning is given against administration of any aluminium-containing medicament or food to subjects with reduced renal function. PMID- 11019593 TI - [Levels of interleukin-4, interleukin-5, tryptase and eosinophil cationic protein of nasal lavage fluid in pollen allergic rhinitis]. AB - IL-4, IL-5, tryptase and eosinophil cationic protein levels were measured in nasal lavage fluid from 15 pollen allergic rhinitis beyond pollen season. Allergy was proved by prick test. There were 15 non allergic children in the control group. Specific nasal allergen provocation was performed on the rhinitic group. Nasal lavage were done before, 1 and 12 hours after the provocation. Before the nasal provocation the ECP and IL-4 levels were significantly higher in the allergic group compared to the non allergic group. The levels of tryptase, ECP and IL-4 rose significantly after the provocation. The results reflect to the possibility of an activated immune status in allergic rhinitis even without the presence of the triggering pollens. After the specific provocation elevated tryptase levels were measured, referring to the activity of the early phase of the I. type hypersensitivity reaction, while the ECP and IL-5 elevation to its late phase. According to our examinations it can be said, that tryptase, ECP and IL-5 might be used to detect the activation of the early and late phases of the IgE mediated hypersensitive reaction. PMID- 11019594 TI - [MR-guided diagnostic and therapeutic interventions. Review and own preliminary experience]. AB - Magnetic resonance (MR) guided diagnostic and therapeutic interventions are relatively new methods in the larger field of radiologic interventions. Good spatial resolution and soft tissue characterisation, multiplanar imaging capability, functional imaging and up-to-date computer systems enable MR imaging to guide a wide range of interventional procedures. Biopsies, punctures and drainages of fluid collections, tumor ablations and vascular interventions can be performed with the help of MRI. MR imaging provides an unique opportunity for planning, guiding and monitoring certain types of intraoperative procedures. The authors outline the basic features of MR intervention and report their own initial experience in interventional procedures in 8 cases for the first time in Hungary. PMID- 11019595 TI - [Time-course of interocular alignment in arteficially esotropic cats]. AB - Strabismus induced by eye-muscle surgery is a widely accepted experimental model of discordant binocular vision. Up to now, no attention was paid to the progress of strabismus in the postsurgical period, although, the grade of the developing strabismus is strongly influenced by the loss of binocularity in the visual system. Following lateral rectus laminotomy of 2-week-old kittens, the Hirschberg method and prisms were used to assess interocular alignment. The angle of deviation was measured weekly. During the first postoperative weeks, cats fixated still alternally, while by the end of the second postnatal month, when compared to healthy controls, operated cats were unable to align their eyes parallelly. They also showed persistent esotropy (+35 +/- 7 delta) that developed with a characteristic time-course. The interocular misalignment in artificially strabismic kittens develop gradually, and does not simply appear after the operation. The nature of this pathological process resembles the normal development of eye-movements in kittens, as well as in primate infants. Therefore, the surgery for strabismus in experimental animals at the earliest time point during the sensitive period are recommended, in order to provide ample time-window window for the effective development of the "strabismic visual system". PMID- 11019596 TI - Electrophysiological instability in the acute phase: prognostic significance of early ventricular fibrillation in acute myocardial infarction. AB - BACKGROUND: In the prognostic stratification of patients affected by AMI is important to evaluate, besides the assessment of left ventricular function and residual ischemia, the presence of electrophysiological instability. METHODS: We have analysed 15 patients all affected by AMI complicated by early ventricular fibrillation. During the hospital phase we evaluated the E.F.% (ECHO) and the presence of late ventricular potentials (SAECG). After hospital discharge we followed up the patients for 6 months. RESULTS: None of the patients died during the hospital phase while the posthospital cardiac mortality was 20%. The three patients dead during the follow-up had an AMI localized in the anterolateral wall of the left ventricle, an E.F.% less than 40% and LVP positive in the hospital phase. Besides the clinical course was complicated by cardiac failure. CONCLUSIONS: We conclude that these three patients are a "high risk profile subgroup" and should be submitted to extensive evaluation with cardiac catheterization, coronary arteriography and programmed ventricular stimulation. PMID- 11019597 TI - Evaluation of haemostatic parameters and circadian variations of the haemostatic system in patients with systemic sclerosis and Raynaud's phenomenon. AB - BACKGROUND: Systemic sclerosis (SSc) is a multisystemic disease characterized by proliferation and swelling of endothelial cells and other disorders. Raynaud's phenomenon (RP) is a disturbance, with unknown pathogenesis, that may be a precursor to SSc. The aim of this study was to investigate possible alterations in the haemostatic system and to examine whether there is a circadian variation in haemostatic variables at the initial stage of SSc. METHODS: In 20 patients with RP (in all patients secondary to SSc) and in 10 controls the levels of thrombomodulin (TM), beta-thromboglobulin (beta-TG), D-dimer (DD), tissue-type plasminogen activator (t-PA) and plasminogen activator-inhibitor (PAI-1) were measured in venous plasma samples taken at 9.00 and 14.00. RESULTS: Only TM levels were found to be higher in patients than in controls. Moreover the PAI-I levels, in the patient group, showed a significant circadian rhythm (with peak values at 9.00). No significant circadian variations for the other parameters were detected. CONCLUSIONS: These data seem to indicate that in patients with RP there is an endothelial damage reflected by a significant elevation of the TM plasma level and a circadian variation in plasma PAI-1, which was higher in the morning. This observation may be an area worth exploring for its importance potential in the knowledge of Raynaud's phenomenon. PMID- 11019598 TI - ATP-MgCl2 treatment after experimental acute myocardial ischemia and reperfusion. AB - BACKGROUND: The effect of ATP-MgCl2 on myocardial metabolism and hemodynamics was investigated in this study. METHODS: Twelve dogs were entered in this research. Six dogs received ATP-MgCl2 and the remaining dogs were considered as controls. The amount of ATP and MgCl2 concentration of this solution is 100 mumol/ml each. The volume administered to the animals during the aortic occlusion is 0.25 ml/kg/hour; in the solution are 100 mumol/ml dose each. The volume administered to the animals during reperfusion is 0.25 ml/kg/hour. The left anterior descending artery was occluded for a period of one hour and the drug was administered during reperfusion. RESULTS: Three hours after reperfusion, cardiac output was 1524 +/- 26 ml/min in the control group and 1638 +/- 47 ml/min in the ATP-MgCl2 group (p < 0.05), pulmonary capillary wedge pressure was 14 +/- 3 in the control group and 8 +/- 2 in the ATP-MgCl2 group. At the same time interval tissue ATP and lactate level was 7 +/- 3, 1.3 +/- 0.4 in the control group and 14 +/- 2, 0.0 +/- 0.2 in the ATP-MgCl2 group respectively (p < 0.05). CONCLUSIONS: In this study we demonstrated that ATP-MgCl2 usage after one hour of arterial occlusion protects the heart from the adverse effects of ischemia/reperfusion. PMID- 11019599 TI - Low serum alpha-1-antitrypsin specific activity in monoclonal gammopathies. AB - BACKGROUND: Serum alpha 1-antitrypsin (alpha 1AT) antigen concentration is elevated in malignancies as the result of acute phase reaction. In the present study, we examined whether the alpha 1AT elevation in monoclonal gammopathies was accompanied by an adequate increase of its functional activity. METHODS: In this case-control study, serum alpha 1AT concentration was measured in 187 ambulatory patients with monoclonal gammopathies and 320 healthy blood donors matched according to sex and age. The alpha 1AT antigen concentration was assayed by immunonephelometry, whereas its functional activity was measured as trypsin inhibitory capacity (TIC). The specific alpha 1AT inhibitory activity (SIA) was calculated, defined as the TIC/antigen concentration ratio. RESULTS: The alpha 1AT antigen concentrations obtained in the patients' samples were very significantly higher as compared with the corresponding values in the control group (mean +/- SD = 134 +/- 41.9% of normal, p < 0.001). However, the TIC values were higher in the patients than in the healthy controls only by 4% (104 +/- 23.8%, p < 0.05). The specific alpha 1AT activity was very significantly lower in the patients, as compared with the controls (p < 0.001), indicating that serum alpha 1AT in monoclonal gammopathies was partially inactive. CONCLUSIONS: As poor correlation between the TIC values and the antigen concentrations was obtained in the patient group as well as the decreased specific alpha 1AT activities, the TIC values in patients with monoclonal gammopathies should be interpreted with caution. PMID- 11019600 TI - Does holiday hypoglycaemia exist? AB - BACKGROUND: To determine whether an excessive, prolonged and, above all, unusual physical exertion could be associated with episodes of mild hypoglycaemia in non insulin-dependent diabetes mellitus (NIDDM) patients treated with glibenclamide. METHODS: EXPERIMENTAL DESIGN: 11 months of observation with retrospective analysis of patient personal diaries to determine the hypoglycaemic risk. SETTING: Diabetic Unit-Department of Medicine and Aging-Chieti University School of Medicine. PATIENTS: We enrolled 340 NIDDM outpatients adjusted for sex, age, body mass index, alcohol intake and oral treatment regimen with glibenclamide. PATIENTS were tested monthly for circadian blood glucose profiles and glycosylated hemoglobin. Mild hypoglycaemia was defined on the basis of blood glucose values < 2.8 mmol/l associated with mild autonomic symptoms, without requiring external assistance. Each diabetic patient filled personal diary indicating the therapy regimen and the characteristics of eventual hypoglycaemic episodes occurring during the observation period. RESULTS: 21.8% of NIDDM patients experienced one or two episodes of mild hypoglycaemia during the observation period. The analysis of the patients' diaries showed that 60% of the hypoglycaemic episodes was associated with excessive, prolonged and unexpected physical exertions. Within this group, about 70% of the episodes occurred during a holiday ("holiday hypoglycaemia"). After analyzing the socio-demographic and clinical characteristics of the diabetic patients reporting hypoglycaemic events, we found a higher risk for "holiday hypoglycaemia" in patients with a lower educational level, with a sedentary occupation or among the ex-farmers. CONCLUSIONS: As resulted in the present study, unexpected physical exertions may represent a relevant cause of mild hypoglycaemia in diabetic patients receiving oral antidiabetic therapy. However, this hypoglycaemic cause may have been underestimated in the literature. Educational programs conducted by general practitioners or diabetologists could be useful for the patients in reducing the number of mild hypoglycaemic episodes. PMID- 11019601 TI - Azoospermia: incidence, and biochemical evaluation of seminal plasma by the differential pH method. AB - BACKGROUND: Recent artificial reproductive technologies (ART) have enabled even azoospermic subjects to take part in "in vitro fertilization" programs. The aim of the present study was: a) to evaluate the incidence of azoospermic male partners of infertile couples; b) to determine whether biochemical evaluation of such azoospermic males can help to discriminate between obstructive and secretory azoospermia. METHODS: Semen samples (n. 28,339; period 1990-1997) were analyzed according to WHO guidelines. In addition, in 108 azoospermic subjects, fructose, citric acid and L-carnitine (taken respectively as markers of seminal vesicle, prostate and epididymal function) were assayed using a new, faster technique, the differential pH method. RESULTS: The incidence of azoospermic semen samples was 4.76%. The biochemical analyses were indicative diagnostically, especially with regard to obstructive azoospermia. However, in secretory azoospermia, biochemical analysis of seminal plasma did not offer particularly helpful indications, except in cases of hypogonadism. CONCLUSIONS: Azoospermia accounts for a relatively high number of cases and justifies detailed attention to the diagnostic and therapeutic approach. Biochemical analysis of the seminal markers is a valid support for differential diagnosis between secretory obstructive azoospermia. It can therefore help in the correct recruitment of such patients for ART programs. PMID- 11019602 TI - Socio-demographic factors and indications in second trimester voluntary abortion. AB - BACKGROUND: According to Italian Law, second trimester termination of pregnancy is allowed for life threatening conditions or for severe psychological distress, linked or not to prenatal diagnosis of foetal abnormalities. Socio-demographic factors related to this condition have been analysed. METHODS: Clinical records of 330 patients admitted during the years 1988-1997 to the Obstetrics and Gynaecology Department, University of Bari, Italy, for voluntary second trimester abortion, were examined. Maternal psychiatric indications have been given in nearly all of the cases. In 123 cases the indications were secondary to the women suffering a psychiatric disorder due to foetal pathologies. In 205 cases--where poor social conditions were more frequent--the indication was given on the ground of a psychiatric disorder linked to the pregnancy itself. RESULTS: Significantly higher incidence of teenagers (23.3%) and singles (50%) in women who underwent a late abortion. Students were 16.4% in this group. In primary psychiatric indication singles prevail (74.4%) and students represent 23.6% while in secondary psychiatric indication the married were 84.7%, students only 4.8%. In primary psychiatric indication 32.5% of women aged nineteen or less, while in secondary psychiatric indication this percentage was 8%. CONCLUSIONS: Among patients who have a late abortion, teenagers students and singles are prevalent, these patients have significantly more primary psychiatric indications, not linked to foetal abnormalities. The high percentage of teenagers with primary psychiatric indication could depend on inadequate information and social service. Reduction of mid-trimester terminations of pregnancy can be significantly achieved intervening in this group of young women. On the other hand, in secondary indications earlier diagnosis of foetal abnormalities must be encouraged (villocentesis instead of amniocentesis) and abortion discouraged when the foetal pathology is minor, treatable or unlikely to significantly impair the future quality of life. PMID- 11019603 TI - Fetuses with cystic hygroma. A retrospective study. AB - BACKGROUND: The aim of this study is to analyse several theories of pathogenesis of cystic hygroma, its correlation with chromosomal abnormalities and the indicators of poor or good prognosis. METHODS: EXPERIMENTAL DESIGN: This is a retrospective study that evaluates all cases of cystic hygroma seen during the four-year period from January 1994 to December 1997. SETTING: This study was performed in the center of prenatal diagnosis of institutional hospital in Caserta. PATIENTS: All cases of fetuses with cystic hygroma were examined in 2100 pregnant female who visited the ambulatory. INTERVENTIONS: The modality of diagnosis of this pathology and the presence of abnormal maternal serum levels of alpha-fetoprotein, human chorionic gonadotropin and unconjugated estriol were evaluated. Moreover, the presence of karyotype abnormalities or other non chromosomal abnormalities were also evaluated. Prognostic indicators such as the presence of septae seen by sonography were examined. RESULTS: Nine fetuses with cystic hygroma were diagnosed sonographically. Septae were identified in six cases. Chromosomal abnormalities were found in five cases. Two cases presented Turner's syndrome and one case Downs' syndrome. There were two cases with associated anomalies. The amniotic fluid alpha-fetoprotein (AFP) levels were high in all cases. CONCLUSIONS: Cystic hygroma is a malformation of the lymphatic system that is diagnosed by ultrasound very well from the first quarter of pregnancy. It is frequently associated with chromosomal and non chromosomal abnormalities. The presence of septae in it and amniotic fluid AFP levels are prognostic indicators. PMID- 11019604 TI - Tamoxifen effects on endometrium. AB - BACKGROUND: To evaluate the effects on the endometrium of a long term treatment with Tamoxifen in postmenopausal patients, asymptomatic for gynecologic disorders, surgically treated for breast cancer. METHODS: SETTING: Outpatient menopausal clinic and endoscopic unit. PATIENTS AND INTERVENTIONS: 45 patients (Group I) were treated with 20 mg of Tamoxifen daily for a mean of 23.4 months. Seven patients (Group II) represented the control group and did not receive Tamoxifen. A transvaginal ultrasonography and a hysteroscopic guided biopsy were performed in all patients. RESULTS: Sagittal sonograms showed abnormal endometrial thickening (range 8-32 mm, mean 13 mm) in 17 patients (35.4%) of Group I and in 1 patient of Group II. Pathology on endometrial tissue sampling obtained at the time of hysteroscopy showed hyperplastic endometrial polyps in 3 patients (6.25%), endometrial hyperplasia in 16 patients (33.4%), while 1 patient had an endometrial polyp cancer on a background of hyperplasia and 1 had a superficial endometrial cancer (4.1%). Out of the 7 patients of Group II, one had an endometrial polyp, while 6 had no relevant endometrial abnormalities. CONCLUSIONS: Our study confirms that Tamoxifen treatment is associated with an increased incidence of proliferative and neoplastic endometrial changes. No obvious correlation was found between the length of Tamoxifen exposure time and occurrence of endometrial pathologies. It is mandatory to undertake twice per year gynecological evaluations for patients treated with Tamoxifen to promptly identify and correctly manage endometrial changes. PMID- 11019605 TI - Evaluation of polyamide synthetic hair. A long-term clinical study. AB - BACKGROUND: Different methods of hair restoration surgery exist: expanders, rotation flaps, autotransplantation and artificial hair implant. METHODS: The authors reviewed 196 patients who underwent hair implantation 2 years before with a highly biocompatible polyamide fiber. Indications, contraindications and complications of the procedure are outlined. RESULTS: Clinical subjective and photographic objective judgement, evaluation of Hamilton scale grading and covered area rate show very satisfactory improvements. Relevant adverse events were limited to 1.02%. CONCLUSIONS: A careful medical follow-up with regular scalp check-up minimizes complications to a very acceptable rate and overall results are definitely satisfying. PMID- 11019606 TI - Role of sucralfate in gastrointestinal diseases. AB - Sucralfate is a cytoprotective drug widely used in clinical practice to prevent or treat several gastrointestinal diseases such as gastro-esophageal reflux, gastritis, peptic ulcer, stress ulcer and dyspepsia. Sucralfate is a safe and well tolerated drug, as demonstrated by the quite complete lack of side effects and it is, for this reason, one of the most important therapeutic choices in the management of acid related diseases during pregnancy. Moreover, sucralfate has recently been shown to be useful in non-acid related gastrointestinal disease as well. In fact, sucralfate has also been administered topically in patients with radiation-induced mucosal procto-sigmoiditis or ulcerative colitis with surprising results. The drug is actually able to form a physical barrier between epithelium and damaging agents (-bile salts, drugs, refluxate...). Moreover, sucralfate increases the local levels of fibroblast growth factors and induces a rise in the mucosal concentration of prostaglandins which are considered important factors in mucosal healing. The aim of this paper is to describe the current and probably forthcoming uses of sucralfate in the field of gastrointestinal disorders. Moreover, we investigate the role of sucralfate as a reliable means to prevent the occurrence of reflux-like symptoms after Helicobacter pylori eradication and in the management of Helicobacter pylori negative patients affected by non-ulcer dyspepsia. PMID- 11019607 TI - Tourette's syndrome in children: neurobiological issues in pathophysiology. AB - In recent decades significant advances in the understanding of neurobiological substrates of Tourette's Syndrome (TS) have led to the formulation of hypotheses regarding the ways in which the most salient features of the syndrome may occur. Pathophysiology of TS involves multiple intertwined neurobiological issues in different areas of the Central Nervous System. This review considers neuroimaging studies (MRI, PET, SPECT) in patients with TS. Neurochemical neurophysiological and electrophysiological studies are also reviewed. The role of the neuroendocrine and neuroimmunologic mechanism on pathogenesis of the disease is discussed. Advances in diagnostic techniques (fMRI, mMRI, PET, SPECT) and in neurophysiological research on neurotransmitter systems will allow us to better understand the pathophysiology of TS and to use more specific treatments. PMID- 11019608 TI - Red radioluminescence and radiochemiluminescence: premises for a photodynamic tumour therapy with X-rays and haematoporphyrin derivatives. A working hypothesis. AB - In a 1989 paper the author put forward the hypothesis of homogeneously illuminating the interior of the human body with red light by means of X-rays, with a view to extending the possibilities of photodynamic therapy with haematoporphyrin derivatives (HPD). In the present paper, two possible mechanisms for the production of red light by tissues irradiated with X-rays are put forward: a physical mechanism (red radioluminescence) and a physical-chemical mechanism (red radiochemiluminescence). Reactive oxygen metabolites produced in the radiolysis of water, singlet oxygen and lipid peroxidation of biomembranes are the key players in this interesting process. However, before any clinical application can be proposed for integrated tumour therapy (photodynamic therapy with X-rays instead of laser light and haematoporphyrin derivatives), quantitative analyses and experimental research will be required. PMID- 11019609 TI - Previous tuberculosis, hepatitis C virus and lichen planus. A report of 10 cases, a causal or casual link? AB - We report 10 cases of lichen planus (LP) and chronic liver disease linked to HCV. The mean age was 63.4 +/- 5.1 years (range 51-73), five were female; six patients had an established cirrhosis of the liver, as shown by either a liver biopsy or the ultrasonographic and biohumoral evidence. The remaining four patients had chronic hepatitis. Histological examination confirmed the presence of LP: the localization of the dermatosis was restricted to the skin in four patients, to the mucous membranes in five (4 atrophic erosive and one erosive) while the remaining had mucous-cutaneous localization. A type II cryoglobulinemia was demonstrated in two and a type III in one of the patients, while no one had otherwise circulating autoantibodies (anti-nuclear, anti-smooth muscle, anti liver kidney microsomal type 1 and anti-mitochondrial antigens) such as other etiological factors of liver disease. In six of the patients the history was positive for previous Mycobacterium tuberculosis infection. In clinical practice the patients with chronic liver disease and HCV infection can also suffer from severe extrahepatic manifestations, including lichen planus. PMID- 11019610 TI - Takayasu's arteritis associated with Crohn's disease. Report of a case. AB - The authors report a rare case of Takayasu's arteritis with aortic arch and abdominal aorta involvement associated with Crohn's disease of the colon which both occurred in a young female patient. The coexistence of two immune mediated diseases in the same subject is unusual for they are generally considered to be independent of each other, however a hypothesis of their possible interrelationship is put forward. PMID- 11019611 TI - History of laparoscopic surgery. AB - Laparoscopic surgery, whose development has been so impressive in the last decade, was initially introduced at the beginning of this century by Dimitri Ott, Georg Kelling and Hans Christian Jacobeus. Von Ott inspected the abdominal cavity of a pregnant women in 1901 and afterwards Georg Kelling performed a procedure, called "koelioscopie", closer to the definition of modern laparoscopy. In the same year Jacobeus published his first report of what he called "Laparothorakoskopie". In the following years several authors in Europe and in the United States performed laparoscopic procedures for diagnostic purposes. It was only with the introduction of the rod-lens optical system and of the cold light fiber-glass illumination that laparoscopy became more popular especially in the gynecologist departments. At this time laparoscopy in general surgery was mainly performed for the diagnosis of liver disorders and abdominal trauma, until the intuition of Lukichev in 1983 and Muhe in 1985 who performed their personal technique of laparoscopic cholecystectomy in humans. Their rudimental techniques did not receive the attention they probably deserved and increasing interest in laparoscopy among general surgeons developed only after the French gynecologist Mouret performed in 1987 the first acknowledged laparoscopic cholecystectomy by means of four trocars. Operative laparoscopy has advanced surprisingly in the last ten years. Several operative procedures have been performed by this new approach. After its tumultuous debut, laparoscopic surgery is now entering a phase of slower development and nowadays it is impossible to predict the immediate and long-term evolution of the technique. PMID- 11019612 TI - The perception of length on curved and flat surfaces. AB - In three experiments, observers judged the apparent extents of spatial intervals along the surface of a curved cylinder or a flat plane that was binocularly viewed in a natural, indoor environment. The observers' judgments of surface lengths were precise and reliable but were also inaccurate and subject to relatively large constant errors. These distortions differed among the observers, but they tended to perceive lengths oriented along the curved dimension of the cylinder as being longer than physically equivalent lengths in the noncurved dimension. This phenomenon did not occur when the observers judged curved and noncurved paths on the flat surface. In addition, some observers' judgments of length were affected by changes in the distance to the cylinder, whereas others were affected by the cylinder's orientation in space. These results demonstrate that the perception of length on surfaces is highly dependent on the particular context in which the length occurs. PMID- 11019613 TI - Heading judgments in minimal environments: the value of a heuristic when invariants are rare. AB - Observers made systematic heading judgments in two experiments simulating their translation through an environment with only two trees. When those trees converged or decelerated apart, observers tended to follow the invariant information and make heading judgments outside the near member of the pair. When those trees accelerated apart, however, observers tended to follow the heuristic information and make judgments outside the far member, although this result was tempered by the angular separation between the trees and their relative acceleration. The simultaneous existence and use of invariants and heuristics are discussed in terms of different metatheoretical approaches to perception. PMID- 11019614 TI - Lightness from contrast: a selective integration model. AB - As has been observed by Wallach (1948), perceived lightness is proportional to the ratio between the luminances of adjacent regions in simple disk-annulus or bipartite scenes. This psychophysical finding resonates with neurophysiological evidence that retinal mechanisms of receptor adaptation and lateral inhibition transform the incoming illuminance array into local measures of luminance contrast. In many scenic configurations, however, the perceived lightness of a region is not proportional to its ratio with immediately adjacent regions. In a particularly striking example of this phenomenon, called White's illusion, the relationship between the perceived lightnesses of two gray regions is the opposite of what is predicted by local edge ratios or contrasts. This paper offers a new treatment of how local measures of luminance contrast can be selectively integrated to simulate lightness percepts in a wide range of image configurations. Our approach builds on a tradition of edge integration models (Horn, 1974; Land & McCann, 1971) and contrast/filling-in models (Cohen & Grossberg, 1984; Gerrits & Vendrik 1970; Grossberg & Mingolla, 1985a, 1985b). Our selective integration model (SIM) extends the explanatory power of previous models, allowing simulation of a number of phenomena, including White's effect, the Benary Cross, and shading and transparency effects reported by Adelson (1993), as well as aspects of motion, depth, haploscopic, and Gelb induced contrast effects. We also include an independently derived variant of a recent depthful version of White's illusion, showing that our model can inspire new stimuli. PMID- 11019615 TI - Nonlinearity in color space measured by apparent motion. AB - We used an apparent motion technique to examine the intensity coding along the three cardinal axes of color space: achromatic (L + M + S), L-M cone, and S cone axes. Two horizontal bars of different colors were alternated to produce a vertical displacement. The color of the background was a mixture varied between the colors of the two bars. When the background color was close to either of the test colors, only the bar that was more salient appeared to jump. Observers adjusted the color of the background until they saw either the two bars moved equally frequently or both bars moved at once. If the color difference in a linear cone excitation space controls this apparent motion, the setting should be midway between the two colors. All of the three cardinal axes showed some deviation from linear behavior. The nonlinearity was less extreme than a logarithmic function for both the achromatic and S cone axes and could be attributed to a small compressive nonlinearity, possibly at the level of cone responses. However, the L-M stimuli showed a more extreme departure from linearity, which suggested a nonlinearity at an opponent site. A test of perceived contrast judgments did not show this nonlinearity for L-M axis, suggesting that it is specific to the L-M contribution to apparent motion. PMID- 11019616 TI - Illusory contours and spatial judgment. AB - We investigated whether, in the human visual system, the mechanisms responsible for relative location judgments are the same when those judgments are made in the context of illusory contours and in the context of mentally joining two points. We asked subjects to align a dot with the oblique contour of an illusory surface or to align a dot with two markers at an oblique orientation. The systematic errors differed in direction for these two conditions. All the systematic errors were orientation dependent. The errors in aligning a dot with an illusory contour seem to be related to the asymmetrical shape of the single objects, which are able to induce an illusory contour, as well as figure-ground segregation. PMID- 11019617 TI - Coming to grips with weight perception: effects of grasp configuration on perceived heaviness. AB - We investigated how changes in grasp configuration affect perceived heaviness in a weight discrimination task in which participants compared the weights of a series of test objects with the weight of a reference object. In different experiments, we varied the width of the grasp, the number of digits employed, the angle of the grasp surface, and the size of the contact area between the digits and the object. We show that objects are perceived to be lighter when lifting with (1) a wide grip in comparison with a narrow grip, (2) five digits in comparison with two digits, and (3) a large contact area in comparison with a small contact area. However, the angle of the contact surfaces did not influence perceived weight. We suggest that changes in central motor commands associated with grasp differences may influence perceived weight, at least under some conditions. PMID- 11019618 TI - The generation of vibrotactile patterns on a linear array: influences of body site, time, and presentation mode. AB - In order to provide information regarding orientation or direction, a convenient code employs vectors (lines) because they have both length and direction. Potential users of such information, encoded tactually, could include persons who are blind, as well as pilots, astronauts, and scuba divers, all of whom need to maintain spatial awareness in their respective unusual environments. In these situations, a tactile display can enhance environmental awareness. In this study, optimal parameters were explored for lines presented dynamically to the skin with vibrotactile arrays on three body sites, with veridical and saltatory presentation modes. Perceived length, straightness, spatial distribution, and smoothness were judged while the durations of the discrete taps making up the "drawn" dotted lines and the times between them were varied. The results indicate that the two modes produce equivalent sensations and that similar sets of timing parameters, within the ranges tested, result in "good" lines at each site. PMID- 11019619 TI - Strategic control over saccadic eye movements: studies of the fixation offset effect. AB - We studied the strategic (presumably cortical) control of ocular fixation in experiments that measured the fixation offset effect (FOE) while manipulating readiness to make reflexive or voluntary eye movements. The visual grasp reflex, which generates reflexive saccades to peripheral visual signals, reflects an opponent process in the superior colliculus (SC) between fixation cells at the rostral pole, whose activity helps maintain ocular position and increases when a stimulus is present at fixation, and movement cells, which generate saccades and are inhibited by rostral fixation neurons. Voluntary eye movements are controlled by movement and fixation cells in the frontal eye field (FEF). The FOE--a decrease in saccade latency when the fixation stimulus is extinguished--has been shown to reflect activity in the collicular eye movement circuitry and also to have an activity correlate in the FEF. Our manipulation of preparatory set to make reflexive or voluntary eye movements showed that when reflexive saccades were frequent and voluntary saccades were infrequent, the FOE was attenuated only for reflexive saccades. When voluntary saccades were frequent and reflexive saccades were infrequent, the FOE was attenuated only for voluntary saccades. We conclude that cortical processes related to task strategy are able to decrease fixation neuron activity even in the presence of a fixation stimulus, resulting in a smaller FOE. The dissociation in the effects of a fixation stimulus on reflexive and voluntary saccade latencies under the same strategic set suggests that the FOEs for these two types of eye movements may reflect a change in cellular activity in different neural structures, perhaps in the SC for reflexive saccades and in the FEF for voluntary saccades. PMID- 11019620 TI - Evidence for auditory feature integration with spatially distributed items. AB - Recent auditory research using sequentially presented, spatially fixed tones has found evidence that, as in vision for simultaneous, spatially distributed objects, attention appears to be important for the integration of perceptual features that enable the identification of auditory events. The present investigation extended these findings to arrays of simultaneously presented, spatially distributed musical tones. In the primary tasks, listeners were required to search for specific cued conjunctions of values for the features of pitch and instrument timbre. In secondary tasks, listeners were required to search for a single cued value of either the pitch or the timbre feature. In the primary tasks, listeners made frequent errors in reporting the presence or absence of target conjunctions. Probability modeling, derived from the visual search literature, revealed that the error rates in the primary tasks reflected the relatively infrequent failure to correctly identify pitch or timbre features, plus the far more frequent illusory conjunction of separately presented pitch and timbre features. Estimates of illusory conjunction rate ranged from 23% to 40%. Thus, a process must exist in audition that integrates separately registered features. The implications of the results for the processing of isolated auditory features, as well as auditory events defined by conjunctions of features, are discussed. PMID- 11019621 TI - On asymmetries in cross-modal spatial attention orienting. AB - In a previous study, Ward (1994) reported that spatially uninformative visual cues orient auditory attention but that spatially uninformative auditory cues fail to orient visual attention. This cross-modal asymmetry is consistent with other intersensory perceptual phenomena that are dominated by the visual modality (e.g., ventriloquism). However, Spence and Driver (1997) found exactly the opposite asymmetry under different experimental conditions and with a different task. In spite of the several differences between the two studies, Spence and Driver (see also Driver & Spence, 1998) argued that Ward's findings might have arisen from response-priming effects, and that the cross-modal asymmetry they themselves reported, in which auditory cues affect responses to visual targets but not vice versa, is in fact the correct result. The present study investigated cross-modal interactions in stimulus-driven spatial attention orienting under Ward's complex cue environment conditions using an experimental procedure that eliminates response-priming artifacts. The results demonstrate that the cross modal asymmetry reported by Ward (1994) does occur when the cue environment is complex. We argue that strategic effects in cross-modal stimulus-driven orienting of attention are responsible for the opposite asymmetries found by Ward and by Spence and Driver (1997). PMID- 11019622 TI - Selective attention to the chemosensory modality. AB - Previous studies have shown that behavioral responses to auditory, visual, and tactile stimuli are modulated by expectancies regarding the likely modality of an upcoming stimulus (see Spence & Driver, 1997). In the present study, we investigated whether people can also selectively attend to the chemosensory modality (involving responses to olfactory, chemical, and painful stimuli). Participants made speeded spatial discrimination responses (left vs. right) to an unpredictable sequence of odor and tactile targets. Odor stimuli were presented to either the left or the right nostril, embedded in a birhinally applied constant airstream. Tactile stimuli were presented to the left or the right hand. On each trial, a symbolic visual cue predicted the likely modality for the upcoming target (the cue was a valid predictor of the target modality on the majority of trials). Response latencies were faster when targets were presented in the expected modality than when they were presented in the unexpected modality, showing for the first time that behavioral responses to chemosensory stimuli can be modulated by selective attention. PMID- 11019623 TI - Object-based selection under focused attention: a failure to replicate. AB - In a recent study, Lavie and Driver (1996) reported that object-based effects found with distributed attention disappear when attention is focused on a narrow area of the display. This finding stands in contrast with previous reports of object-based effects under conditions of focused attention (e.g., Atchley & Kramer, 1998; Egly, Driver, & Rafal, 1994). The present study was an attempt to replicate Lavie and Driver's finding, using similar task and stimuli. While Lavie and Driver's object-based effect in the distributed attention condition was replicated, its absence in the focused attention condition was not. In the two experiments reported in this paper, object-based effects were found under conditions of both distributed and focused attention, with no difference in the magnitude of the object-based effects in the two conditions. It is concluded that, in contrast with Lavie and Driver's claim, the initial spatial setting of attention does not influence object-based constraints on the distribution of attention. PMID- 11019624 TI - Attending, ignoring, and repetition: on the relation between negative priming and inhibition of return. AB - A series of spatial localization experiments is reported that addresses the relation between negative priming and inhibition of return. The results of Experiment 1 demonstrate that slowed responses to repeated location stimuli can be obscured by repetition priming effects involving stimulus dimensions other than spatial location. The results of Experiments 2, 3A, and 3B demonstrate that these repetition priming effects may occur only when participants are required to respond to the prime display. Together, these results suggest that differences between attended and ignored repetition effects in selective attention studies of spatial localization do not provide a basis for distinguishing between spatial negative priming and inhibition of return. PMID- 11019625 TI - Effects of subphonetic and syllable structure variation on word recognition. AB - This study explored whether natural acoustic variations as exemplified by either subphonetic changes or syllable structure changes affect word recognition processes. Subphonetic variations were realized by differences in the voice-onset time (VOT) value of initial voiceless stop consonants, and syllable structure variations were realized by vowel deletion in initial unstressed syllables in multisyllable words. An auditory identity priming paradigm was used to determine whether the amount of facilitation obtained to a target stimulus in a lexical decision task was affected by the presence of these acoustic variations in a prime stimulus. Results revealed different patterns for the two types of variability as a function of lexical status. In the case of subphonetic variations, shortening of VOT resulted in reduced facilitation for words but not for nonwords, whereas in the case of syllable structure variation, vowel deletion in an unstressed syllable resulted in reduced facilitation for nonwords and increased facilitation for words. These findings indicate that subphonetic variability interferes with word recognition, whereas syllable structure variability does not, and that this effect is independent of the magnitude of the acoustic difference between a citation form and its variant. Furthermore, the results suggest that the lexical status of the target item plays a crucial role in the processing of both types of variability. Results are considered in relation to current models of word recognition. PMID- 11019626 TI - Differences in American English, Spanish, and monkey perception of the say-stay trading relation. AB - An interesting phenomenon in human speech perception is the trading relation, in which two different acoustic cues both signal the same phonetic percept. The present study compared American English, Spanish, and monkey listeners in their perception of the trading relation between gap duration and F1 transition onset frequency in a synthetic say-stay continuum. For all the subjects, increased gap duration caused perception to change from say to stay; however, subjects differed in the extent to which the F1 cue traded with gap duration. For American English listeners, a change from a low to a high F1 onset caused a phoneme boundary shift of 26 msec toward shorter gap durations, indicating a strong trading relation. For Spanish listeners, the shift was significantly smaller at 13.7 msec, indicating a weaker trading relation. For monkeys, there was no shift at all, indicating no trading relation. These results provide evidence that the say-stay trading relation is dependent on perceptual learning from linguistic exposure. PMID- 11019627 TI - [Bedside biology of illness in neonatology]. PMID- 11019628 TI - [The broadcasting physician]. PMID- 11019629 TI - [Helicobacter pylori infection]. PMID- 11019630 TI - [Bacteriology and pathogenicity of Helicobacter pylori]. AB - Helicobacter pylori is the prototype of bacteria belonging to a new genus, the Helicobacter genus. It is a gram-negative, highly motile and microaerophilic bacterium, with a spiral shape, that colonizes the human gastric mucosa and causes several gastroduodenal diseases. Pathogenicity of H. pylori relies upon its capacity to adapt to a hostile environment and to escape the host response. Resistance to acidity, motility, adhesion, molecular mimicry, resistance to phagocytosis, synthesis of a cytotoxin, induction of an inflammatory response are the major strategies developed by H. pylori to colonize persistently and damage gastric tissue. PMID- 11019631 TI - [Epidemiology, acquisition and transmission of Helicobacter pylori]. AB - Helicobacter pylori infection is a chronic infection essentially acquired during childhood. Its prevalence in developed countries like France has decreased according to the year of birth all along the twentieth century, reflecting the progressive improvement of socio-economic and environmental conditions. The incidence of acquisition in adulthood is lower than 0.5% per year. In developing countries, the prevalence is still very high, even in early childhood. H. pylori is a strictly human bacterium. Its reservoir is essentially the stomach. Transmission most likely occurs between humans by an oro-oral or a gastro-oral transmission. Faeco-oral transmission, either direct or via the environment, is possible but seems to be rare in France. PMID- 11019632 TI - [Diagnosis of Helicobacter pylori infection]. AB - Direct or indirect methods are available to diagnose Helicobacter pylori infection. Direct assessment with histopathology, culture, rapide urease test, amplification of gene sequences requires the collection of biopsy samples via an endoscope. Culture is the gold standard, but histopathology is commonly used in clinical practice. Serology, 13C-urea breath test and detection of antigens in stools are indirect methods. In clinical practice, direct assessment is required for the initial diagnosis as only patients with endoscopic lesions need H. pylori eradication. Among indirect methods, serology should be used only in epidemiologic studies. Urea breath test and detection of antigens in stools are the most appropriate methods to assess eradication. PMID- 11019633 TI - [Gastroduodenal ulcer associated with Helicobacter pylori]. AB - Helicobacter pylori is associated with chronic peptic ulcer disease. The high prevalence of H. pylori infection tends to decrease in Western countries suggesting that the incidence of gastroduodenal ulcers not related to antiinflammatory drugs will fall. Abundant data support the leading role of H. pylori in peptic disease through dysregulation of acid production, induction of gastric metaplasia, and development of chronic gastritis. Endoscopy is the key to diagnosis, allowing direct assessment of H. pylori infection. H. pylori eradication is the major aim of peptic ulcer treatment, dramatically improving the recurrence of ulcers, although its efficiency should be controlled in various clinical situations. PMID- 11019634 TI - [Helicobacter pylori and gastric neoplasms: adenocarcinoma and lymphoma]. AB - Helicobacter pylori is a risk factor for gastric carcinoma and an established carcinogenic bacterium. The relative risk to induce a gastric cancer is estimated to be 3 to 6 compared to that of individuals without H. pylori. Gastric atrophy and intestinal metaplasia in gastric mucosa are 2 well recognized precancerous lesions. Their occurrence and evolution are multifactorial depending on age at first infection, duration of infection and host's genetic characteristics. Prevention using H. pylori eradication is recommended only in individuals with high risk of cancer. Gastric lymphoma, although less frequent, may be due to H. pylori infection. Only in low grade lymphoma H. pylori eradication and periodic surveillance are recommended. PMID- 11019635 TI - [Gastritis, dyspepsia and Helicobacter pylori]. AB - Helicobacter pylori is a major culprit in chronic gastritis. Interactions between the infected host, the bacteria and the environment, influence the type of gastritis and the occurrence of specific diseases (peptic ulcer, gastric lymphoma and adenocarcinoma) associated with the infection in a minority of cases. The benefit of H. pylori eradication is not proved in isolated gastritis, except for rare forms such as hypertrophic or lymphocytic gastritis. In spite of a high prevalence in functional dyspepsia with normal gastroduodenal endoscopy, H. pylori is not the main cause of symptoms in dyspeptic patients, but the gastritis could be involved in the appearance of ulcer-like symptoms. Strategies for non invasive H. pylori detection for primary eradication are developed to reduce the endoscopy workload in the management of uninvestigated dyspepsia. PMID- 11019636 TI - [Helicobacter pylori infection in children]. AB - Helicobacter pylori infection is frequent in children. Its incidence in Europe, around 6% in children aged 6-16 years, varies with the socio-economic level and nutritional status. It may reach 46% in Africa and up to 75% in some institutions. Clinical manifestations debated. Vomiting, dyspepsia and acute pain related to ulcer disease may undisputedly be linked to H. pylori, whereas its role in chronic abdominal has yielded contradictory reports. Direct isolation of the bacterium is classically done through perendoscopic antral biopsies followed by culture and histology. Non-invasive diagnosis methods get a wider use in children. Serodiagnosis is reproducible and easy only in older children. The 13C urea breath test is sensitive and specific, and seems perfectly suitable in pediatrics. The H. pylori stool antigen test for the detection of infection seems promising but not yet of current clinical use. Triple therapy using amoxicillin clarithromycin (or metronidazole or tinidazole) and anti-secretory agents is recognised as the most efficient association. PMID- 11019637 TI - [Treatment of Helicobacter pylori infection]. AB - The recommended treatment of Helicobacter pylori infection is a seven-day course of the following regimen: a double dose proton-pump inhibitor (omeprazole 2 x 20 mg, or lansoprazole 2 x 30 mg, or pantoprazole 2 x 40 mg), associated with 2 antibiotics (amoxicillin 2 x 1,000 mg, clarithromycin 2 x 500 mg). Eradication rate is around 65%. Main causes of failure are antibiotic resistance, bad compliance, early termination of therapy due to adverse events. A second line treatment is possible with substitution of clarithromycine by metronidazole (3 x 500 mg), a longer duration of treatment (14 days) and sometimes a quadritherapy with bismuth (not available in France). PMID- 11019638 TI - [France 1]. PMID- 11019639 TI - [Knee pain. Diagnostic orientation]. PMID- 11019640 TI - [Social protection. Principles of social risk in the management of social security, financing mechanisms for health expenditures]. PMID- 11019641 TI - [Cellular and extracellular dehydration and hyperhydration. Etiology, physiopathology, diagnosis, treatment]. PMID- 11019642 TI - [Insulin-dependent diabetes. Etiology, physiopathology, diagnosis, complications, treatment]. PMID- 11019643 TI - [Seizures in the neonate. Diagnostic orientation and emergency guidelines with medical posology]. PMID- 11019644 TI - [Obliteration of the central vein of the retina and its branches. Etiology, diagnosis, development, principles of treatment]. PMID- 11019645 TI - [Immunotolerance and autoimmunity]. PMID- 11019646 TI - [Development of trace metal ion analysis]. AB - Analyses of trace biologically essential or toxic ionic compounds found in the environment are very important. However, the lack of sensitivity and interference caused by coexisting components are often serious problems. To determine trace levels of metal ions without the above problems, new preconcentration and analytical methods have been developed. Firstly, three methods for the selective preconcentration of metal ions are shown below: 1) 3-Chloropyridazine-6 carbohydrazide was immobilized on glass beads supports to be used as a column packing material. Multi-metal ions were concentrated on the column and eluted selectively with several buffers and hydrochloric acid. The eluate was analyzed off-line by flame atomized-atomic absorption spectrometry (AAS). This method was able to determine sub-ppb levels of cupper- and cadmium-ions in environmental samples. 2) Salicylideneamino-2-thiophenol was immobilized on the supports. Aluminum ion was concentrated selectively on the column and eluted with nitric acid. The eluate was analyzed off-line by flameless-AAS or on-line by flow injection analysis using pyrocatechol violet for a post-column colorimetric reagent. These methods were able to determine ppb-ppt levels of aluminium in environmental samples and were suitable for its state-analysis. 3) Bathocuproinesulfonic acid was immobilized on the supports. Copper ion was concentrated selectively on the column and eluted with nitric acid. The eluate was analyzed on-line by flow injection analysis using bathocuproinesulfonic acid. This method was able to determine sub-ppb levels of copper in environmental samples. On the other hand, to analyze simultaneously trace metal ions and anions, capillary electrophoresis was performed using ethylenediaminetetraacetic acid as an electrolyte component. Simultaneous determination of several ions in mineral waters was achieved by the system. PMID- 11019647 TI - [Organometal-catalyzed asymmetric synthesis stereocontrolled by organosulfur functionality]. AB - Our recent studies on asymmetric synthesis with the assistance of organometal catalysts stereocontrolled by organosulfur functionality involved in reaction substrates or ligands are reviewed. The studies focused on asymmetric synthesis via chiral pi-allylmetal complexes derived from (S)-proline allyl ester, olefinic cyclopropanes, chiral 2-alkynyl sulfinates, and chiral new ligands, and also focused on asymmetric cycloaddition reactions with chiral sulfoxides and sulfinates, such as intramolecular ene, metallo-type ene, and hetero Diels-Alder reactions. Participation of organosulfur functionality in organometal-catalyzed asymmetric reactions was unveiled on the basis of the stereochemical outcomes obtained. PMID- 11019648 TI - [Pharmacognosical study on secondary metabolites]. AB - Clonal micropropagation on various medicinal plants was set up resulting in the regenerated plants which possessed a homogeneous quality. The ratio of hapten to bovine serum albumin (BSA) in an antigen conjugate was determined by matrix assisted laser desorption/ionization of mass spectrometry. A hybridoma secreting monoclonal antibody (MAb) was produced by fusing splenocytes immunized with an antigene-BSA conjugate with mouse myeloma cells. Competitive enzyme-linked immunosorbent assay (ELISA) using MAb was set up as a high sensitive, specific and reproducible qualitative method. A method of determination for ginsenosides by using a unique western blotting was established. Immunoaffinity column chromatography using an anti-ginsenoside Rb1MAb has made possible a single-step separation of ginsenoside Rb1 from a crude ginseng extract. Single chain Fv gene of anti-forskolin MAb was prepared from mRNA of hybridoma secreting anti forskolin MAb and cloned. Gene was constructed into a pET-28a(+) vector producing a scFv protein. Modeling of forskolin and scFV was investigated. THCA synthase was purified from the homogenate of Cannabis sativa leaves on successive column chromatographies. THCA synthase was confirmed to be homogeneity having 75 kDa. To obtain the corresponding cDNA clone of THCA synthase, a set of degenerate promers was constructed based on N-terinal and internal amino acid sequences of THCA synthase. The 5' and 3' ends of cDNA were amplified by RACE. A full sequencing has been determined to be corded a polypeptide having 545 amino acid residues. The cDNA clone was expressed in yeast system via PUC19 vector resulting in THCA synthase activity. PMID- 11019649 TI - [Development and evaluation of a local area network system to make drug information paper for inpatient--adaptation to characteristics of diseases, and expansibility to other hospitals and the Internet]. AB - We developed and evaluated a local area network system to make drug information papers. Because pharmacists should explain drug information both orally and using papers to patients in response to their understanding and characteristics of their diseases. The merit of this system is as follows: pharmacists can use it without any education for its operation; most data can be inputted by selection from the lists and automatic reference; the data are adjustable for individuals; different contents are available for different wards; multiple users can access the same data file. Input data in this system are available for making explanation papers on the medical examination of outpatients, and making a plan for pharmaceutical care and guidance services. This system is also available in other hospitals, and on the internet. First, we surveyed the time required for preparing drug information papers. Making a hand written paper for typical patient takes 395 sec. for manuscript, 160 sec. with the system using text data only, and 178 sec. with the system using text and picture data. Next, we surveyed 27 patients' views on the addition of drug pictures to drug information papers, and with the result that 19 patients thought it very good. Last, we surveyed 13 ward pharmacists about their usage of this system. We found that 8 pharmacists used frequently this system, and 3 pharmacists used the same file in the same hours at the maximum. PMID- 11019650 TI - [Analysis of natural medicines by micellar electrokinetic chromatography (MEKC)- determination of calycosin in Astragali radix]. AB - We previously reported a HPLC identification method for Astragali Radix and its fluid extract using calycosin as a marker substance. However, it took about 40 min for one run using gradient elution. Therefore, we alternatively employed capillary electrophoresis (CE) for the analysis of calycosin in those materials. As a result, calycosin was clearly separated from the other components in Astragali Radix and its fluid extract by micellar electrokinetic chromatography (MEKC) mode with sodium dodecyl sulfate (SDS) within 5 min. Then we determined the content of calycosin in a drinkable preparation. Validation for the developed method was also performed in accordance with the ICH guideline. PMID- 11019651 TI - [Effect of thiabendazole (TBZ) on glutathione (GSH) and GSH related enzymes in mice liver]. AB - The present study examines the effects of thiabendazole (TBZ), its metabolites, 5 hydroxythiabendazole (5-OH TBZ) and 2-acetylbenzimidazole (ABI), and structural related compounds, thiazoles and thioamides on glutathione (GSH) concentration and GSH-related enzymes in the livers of ICR 11 week-old female mice. GSH concentration in liver and kidney of mice given orally TBZ 0.65 mol/kg (TBZ group) increased significantly compared with control mice from 24 h to 48 h after administration of TBZ. Even in mice to which TBZ at 0.175 mol/kg was administered in combination with L-buthionine sulfoximine (BSO) 4 mmol/kg (i.p.) (BSO-TBZ group), kidney GSH showed significant increase compared with BSO-control mice 48 h after the administration of TBZ. gamma-Glutamylcysteine synthetase (gamma-GCS) activity in the livers of the TBZ group markedly increased at 48 h and that of BSO-TBZ group increased from 24 h to 48 h. gamma-GCS in mice liver is thus enhanced by TBZ regardless of BSO administration. Hepatic glutathione peroxidase activity of the TBZ group did not change in response to cumene hydroperoxide assubstrate. That of BSO-treated mice decreased by TBZ-coadministration and significant differences was noted between BSO-control and BSO-TBZ group from 1 h to 48 h later. Hepatic glutathione S-transferase (GST) activity toward 1,2 dichloro-4-nitrobenzene (DCNB) was significantly elevated 24 h after administrations of TBZ in TBZ and BSO-TBZ groups. GST activity toward 1,2-epoxy-3 (p-nitrophenoxy) propane of TBZ group increased from 0.5 h to 24 h. Hepatic GST activity toward DCNB and 1-chloro-2,4-dinitrobenzene did not change by administration of 0.65 mol/kg 5-OH TBZ or ABI but increased by administrations of 0.33 mol/kg of thiazole, 4-methylthiazole, 4,5-dimethylthiazole or 2,4 dimethylthiazole. Increase in GSH concentration and GST activity in mice liver by TBZ administration may be considered to provide protection from TBZ or its active metabolites. PMID- 11019652 TI - [Determination of antimicrobial, 10,10'-oxybis-10H-phenoxarsine, in artificial leather used for the seat of chairs]. AB - Recently, a case of patients with allergic contact dermatitis caused by the poly (vinyl chloride) (PVC) seat, containing 10,10'-oxybis-10H-phenoxarsine (OBPA), of a chair was reported. OBPA was developed as an antimicrobial for plastics such as PVC and polyurethane, and it has been widely used in artificial leather for the seats of chairs and sofas. To identify causative chemicals for allergic contact dermatitis, a combination of patch testing in the patients and chemical analysis of causative products is valuable. However, no analytical method and data of OBPA in commercial products was reported. In this study, a method for the determination of OBPA in the artificial leather (PVV base) was developed. OBPA was extracted from PVC samples with methanol. The extract was loaded on an aluminum oxide column, and washed with diethyl ether:hexane, and eluted with ethanol:hexane. The eluate was evaporated, dissolved in methanol and injected to a HPLC equipped an ODS column and an UV detector (detection wavelength 300 nm). OBPA standard crystal was isolated from commercial agents containing OBPA. The calibration curve for OBPA was linear in the range of 0.1-100 micrograms/ml. The minimum detection and determination concentrations of OBPA in samples were 0.07 and 0.25 microgram/g. By this method, eight PVC sheets for the seat of a chair were analyzed. In two PVC sheets, 52.7 and 84.9 micrograms/g of OBPA were detected. In the PVC product that caused contact dermatitis, OBPA was not found. PMID- 11019653 TI - [Effect of KW-3902, a selective adenosine A1-receptor antagonist, on accumulation of gentamicin in the proximal renal tubules in rats]. AB - Adenosine A1-receptor antagonists have been previously shown to possess protective effects in several nephrotoxic models of acute renal failure. To investigate the mechanism of protective effects of adenosine A1-receptor antagonists, we determined effects of 8-(noradamantan-3-yl)-1,3-dipropylxanthine (KW-3902), a selective adenosine A1-receptor antagonist, on the accumulation of gentamicin (GM) into proximal renal tubules in rats. GM was intravenously administered at a dose of 10 mg/kg in anesthetized rats. KW-3902 (0.1 mg/kg) or its vehicle was given as a bolus injection 6 min before the GM injection. Administration of GM and KW-3902 did not affect the systemic blood pressure, heart rate, renal blood flow and glomerular filtration rate. KW-3902 did not affect the plasma concentration of GM. GM was accumulated in the proximal tubules 4 h after the GM injection. Treatment with KW-3902 significantly decreased the accumulation of GM into the proximal tubules. These results suggest that endogenous adenosine may accelerate the uptake of GM at the proximal tubules, and that the renoprotective effects of the adenosine A1-receptor antagonist may result from inhibiting this action of endogenous adenosine, leading to the suppression of intrarenal accumulation of GM. PMID- 11019655 TI - The use of albumin preparations during surgery. AB - New guidelines for the clinical use of blood preparations intended to promote more rational use were issued by the Japanese Ministry of Health and Welfare in June 1999. The purpose of this article is to clarify the current situation in the use of albumin (Alb) preparations during surgery and to design a plan to promote the rational use of these products at Yamaguchi University Hospital. Of the patients administered Alb preparations during surgery in our hospital over the 6 month period from January 1 to June 30, 1999, 158 were selected based on prescription records of plasma component preparations. Most of the patients (63%) were 60 years old or over. The total amount of Alb administered to the patients was 6150 g. Those patients, whose postoperative serum Alb concentration was lower than 3, 3 to 3.5, 3.5 to 4, and more than 4 g/dl, numbered 60, 45, 21, and 23, respectively. The total amount of Alb overdosed to the patients, whose postoperative Alb concentration was more than 3 g/dl, was 2282 g (37% of the administered Alb). The overdosed patients mainly belonged to the Cardiovascular Surgery Division and were treated with a pump-oxygenator during surgery. Three bottles (37.5 g) of concentrated Alb had usually been used to prime the perfusate. These results indicate that it is possible to cut back on Alb by reconsidering the criteria for its administration. PMID- 11019654 TI - [Effect of itraconazole on digoxin clearance in patients with congestive heart failure]. AB - We showed a digoxin-itraconazole interaction in three patients in whom digoxin serum concentrations were increased. Their electrocardiograms revealed arrhythmias such as ventricular premature contraction, atrioventricular block, and ST depression. The elimination half-life of digoxin in case 3 patient who continued itraconazole therapy was 8.4 days, which was estimated by nonlinear least squares method from the serum concentrations of digoxin versus time curve. In order to evaluate the influence of itraconazole on pharmacokinetic parameters of digoxin, we estimated digoxin clearance by the Bayesian method using the population pharmacokinetic parameters in Japanese patients. During the concomitant use of itraconazole and digoxin, the digoxin clearance in all patients decreased to 50.5 +/- 8.8% (mean +/- S.D.) of the clearance without itraconazole. When digoxin and itraconazole are used concomitantly, careful monitoring of digoxin serum concentrations is necessary. Based on our results of digoxin clearance evaluation, the dose of digoxin should be reduced to 50% of original dose after itraconazole is started, and digoxin serum concentration might be controlled at the same level before the concomitant use. PMID- 11019656 TI - New-hip concerns. PMID- 11019657 TI - Preventing sudden cardiac death. PMID- 11019658 TI - What is atherosclerosis? PMID- 11019659 TI - Second opinions. PMID- 11019660 TI - Prostate surgery follow-up. PMID- 11019661 TI - Blocking shingles pain. PMID- 11019662 TI - Timing breast cancer surgery. PMID- 11019663 TI - ECT helps depressed seniors. PMID- 11019664 TI - Prevent bone loss during steroid therapy. PMID- 11019665 TI - Thalidomide shows promise as cancer treatment. PMID- 11019666 TI - Sex after hysterectomy. PMID- 11019667 TI - Can I safely use powdered fiber drinks such as Metamucil to relieve constipation without my body becoming reliant on them? PMID- 11019668 TI - Why are some medicines taken with food and others on an empty stomach? PMID- 11019669 TI - My doctor recently recommended that I take low-dose aspirin to minimize the risk of a heart attack or stroke. I routinely take ibuprofen for arthritis pain. Do I need both, and is it safe to combine the two? PMID- 11019670 TI - Hepatitis C: an uncertain prognosis. PMID- 11019671 TI - Another diabetes dilemma. PMID- 11019672 TI - Dealing with diverticulitis. PMID- 11019673 TI - HRT and breast cancer. PMID- 11019674 TI - Keeping dry after prostate surgery. PMID- 11019675 TI - Stroke risk from uncontrolled high BP. PMID- 11019676 TI - Treating early breast cancer. PMID- 11019677 TI - Lyme disease: lasting effects? PMID- 11019678 TI - Hair loss and heart disease. PMID- 11019680 TI - Exercise for osteoarthritis. PMID- 11019679 TI - Making the most of mammography. PMID- 11019681 TI - Is there anything you can do after stopping smoking to help your lungs get healthy faster? PMID- 11019682 TI - Why is chickenpox more serious in adults than in children? PMID- 11019683 TI - In an article last summer, you mentioned that a 0.3 milligram daily dose of estrogen is effective in preventing osteoporosis. Is a dose that low also effective for preventing heart disease? PMID- 11019684 TI - Kidney failure. Early detection is key. PMID- 11019685 TI - Health tips. Dial 911.... PMID- 11019686 TI - Two new drugs join the war on superbugs. PMID- 11019687 TI - Elbow surgery for arthritis. Relief when other therapies don't work. PMID- 11019689 TI - Anger. Mad can be bad for your health. PMID- 11019688 TI - Scleroderma. It can damage skin and much more. PMID- 11019690 TI - My wife says I should get a flu shot. I'm only 58. Aren't flu shots recommended for people 65 and older? PMID- 11019691 TI - My friend says the peanut butter she buys is healthier than mine. Is that possible? PMID- 11019692 TI - Management of chronic hypertension during pregnancy. PMID- 11019693 TI - Federal, industry initiatives directed at violative residues in market beef, dairy cows. PMID- 11019694 TI - Pork program seeks to quiet public fear, increase foreign competitiveness. PMID- 11019695 TI - Pawspice an option for pets facing the end. PMID- 11019696 TI - Regulation of pet sales and private breeders. PMID- 11019697 TI - Demographic changes in veterinary medicine. PMID- 11019698 TI - Feminization of veterinary medicine. PMID- 11019699 TI - Feminization of veterinary medicine. PMID- 11019700 TI - Thoughts on anesthetic systems used in small animal practices. PMID- 11019701 TI - Questions tryptophan dosage and pain tolerance. PMID- 11019702 TI - What is your diagnosis? Keratoma in a horse's hoof. PMID- 11019704 TI - When the government needs your property--eminent domain. PMID- 11019703 TI - "It was the best of times, it was the worst of times". A perspective on the KPMG study. PMID- 11019705 TI - Motivations for and barriers to engaging in continuing veterinary medical education. AB - OBJECTIVE: To identify reasons for engagement in continuing veterinary medical education (CVME) activities, obstacles to participation, ways to provide more effective programs, and ideas to improve participation in CVME. DESIGN: Focus group interviews. SAMPLE POPULATION: Selected practicing veterinarians from a state that did mandate continuing education for relicensure. PROCEDURE: 12 focus group interviews were held throughout the state of California between May and September 1998. Practitioners were asked to respond to questions about where they obtain information to improve their practice, what value they see in CVME, what motivates them to participate, what obstacles to CVME participation exist, and ways CVME providers and practitioners could overcome those obstacles. RESULTS: 84 practitioners participated in the focus group interviews. In addition to the educational value of CVME, participation was used to rejuvenate practice life and prevent feelings of isolation. Continuing education activities ranged from problem-oriented chats with colleagues to formal educational programs. Timing of events, distance, money, solo practice, stage of career, and family demands were identified as barriers to participation. Designing and marketing CVME with specific learning objectives and for specific career stages and using new educational delivery technologies were suggested to overcome some of these barriers. CONCLUSIONS AND CLINICAL RELEVANCE: If CVME is to improve practice and patient care, it should be integrated into a practice's strategic planning and considered a legitimate business expense. Decisions about CVME participation are made easier if program objectives are clearly outlined. PMID- 11019706 TI - Owner assessment of the outcome of total hip arthroplasty in dogs. AB - OBJECTIVE: To evaluate owners' perceptions of the outcomes of dogs that have undergone total hip arthroplasty (THA). DESIGN: Survey. SAMPLE POPULATION: Owners of 145 dogs that had undergone THA. PROCEDURE: Surveys were mailed to owners of 353 dogs that underwent THA between 1982 and 1996. Owners were asked to answer multiple-choice questions and provide written comments. RESULTS: The response rate was 41%. For owners who responded to the survey, time from surgery to completion of the questionnaire ranged from 6 months to 11 years. Overall, 122 respondents (84.1%) rated results of THA in their dog as excellent or good. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that owners are generally satisfied with the results of THA in their dogs. PMID- 11019707 TI - Evaluation of transdermal fentanyl patches for analgesia in cats undergoing onychectomy. AB - OBJECTIVE: To evaluate efficacy and safety of using transdermal fentanyl patches (TFP) for analgesia in cats undergoing onychectomy. DESIGN: Randomized controlled clinical trial. ANIMALS: 45 client-owned cats weighing > or = 2.7 kg (5.9 lb) undergoing onychectomy, onychectomy and ovariohysterectomy, or onychectomy and castration. PROCEDURE: Cats were randomly assigned to be treated with a TFP (25 micrograms/h) or butorphanol; TFP were applied a minimum of 4 hours before surgery (approx 8 hours prior to extubation). Rectal temperature, heart rate, respiratory rate, force applied by the forelimbs, and serum fentanyl concentration were measured, and temperament, recovery, degree of sedation, severity of pain, severity of lameness, and appetite were scored before and periodically for up to 40 hours after surgery. RESULTS: Cats treated with a TFP had better recovery scores at 2 of 4 evaluation times, lower sedation scores at 2 of 8 evaluation times, and lower pain scores at 6 of 8 evaluation times, compared with cats treated with butorphanol. Use of a pressure-sensitive mat to evaluate force applied by the forelimbs did not reveal any differences between groups but did reveal a significant difference between preoperative and postoperative values. Mean +/- SD serum fentanyl concentrations were 1.56 +/- 1.08, 4.85 +/- 2.38, 4.87 +/- 1.56, and 4.35 +/- 2.97 ng/ml approximately 8, 24, 32, and 48 hours, respectively, after TFP placement. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that use of a TFP (25 micrograms/h) for postoperative analgesia in cats undergoing onychectomy with or without surgical sterilization is safe and effective. PMID- 11019708 TI - Clinical use of serum parvovirus and distemper virus antibody titers for determining revaccination strategies in healthy dogs. AB - OBJECTIVE: To assess whether serum canine parvovirus (CPV) and canine distemper virus (CDV) antibody titers can be used to determine revaccination protocols in healthy dogs. DESIGN: Case series. ANIMALS: 1,441 dogs between 6 weeks and 17 years old. PROCEDURE: CPV and CDV antibody titers in serum samples submitted to a commercial diagnostic laboratory were measured by use of indirect fluorescent antibody (IFA) tests. On the basis of parallel measurements of CPV and CDV serum antibody titers in 61 paired serum samples determined by use of hemagglutination inhibition and serum neutralization methods, respectively, we considered titers > or = 1:5 (IFA test) indicative of an adequate antibody response. RESULTS: Age, breed, and sex were not significantly associated with adequate CPV- or CDV specific antibody responses. Of 1,441 dogs, 1,370 (95.1%) had adequate and 71 (4.9%) had inadequate antibody responses to CPV, whereas 1,346 of 1,379 (97.6%) dogs had adequate and 33 (2.4%) had inadequate responses to CDV. Vaccination histories were available for 468 dogs (468 for CPV, 457 for CDV). Interval between last vaccination and antibody measurement was 1 to 2 years for the majority (281/468; 60.0%) of dogs and 2 to 7 years for 142 of 468 (30.3%) dogs. Interval was < 1 year in only 45 of 468 (9.6%) dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of adequate antibody responses (CPV, 95.1%; CDV, 97.6%) in this large population of dogs suggests that annual revaccination against CPV and CDV may not be necessary. PMID- 11019709 TI - Effects of butorphanol and carprofen on the minimal alveolar concentration of isoflurane in dogs. AB - OBJECTIVE: To evaluate the effects of butorphanol and carprofen, alone and in combination, on the minimal alveolar concentration (MAC) of isoflurane in dogs. DESIGN: Randomized complete-block crossover study. ANIMALS: 6 healthy adult dogs. PROCEDURE: Minimal alveolar concentration of isoflurane was determined following administration of carprofen alone, butorphanol alone, carprofen and butorphanol, and neither drug (control). Anesthesia was induced with isoflurane in oxygen, and MAC was determined by use of a tail clamp method. Three hours prior to induction of anesthesia, dogs were fed a small amount of canned food without any drugs (control) or with carprofen (2.2 mg/kg of body weight [1 mg/lb]). Following initial determination of MAC, butorphanol (0.4 mg/kg [0.18 mg/lb], i.v.) was administered, and MAC was determined again. Heart rate, respiratory rate, indirect arterial blood pressure, endtidal partial pressure of CO2, and saturation of hemoglobin with oxygen were recorded at the time MAC was determined. RESULTS: Mean +/- SD MAC of isoflurane following administration of butorphanol alone (1.03 +/- 0.22%) or carprofen and butorphanol (0.90 +/- 0.21%) were significantly less than the control MAC (1.28 +/- 0.14%), but MAC after administration of carprofen alone (1.20 +/- 0.13%) was not significantly different from the control value. The effects of carprofen and butorphanol on the MAC of isoflurane were additive. There were not any significant differences among treatments in regard to cardiorespiratory data. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of butorphanol alone or in combination with carprofen significantly reduces the MAC of isoflurane in dogs; however, the effects of butorphanol and carprofen are additive, not synergistic. PMID- 11019710 TI - Monoclonal immunoglobulin G cryoglobulinemia and multiple myeloma in a domestic shorthair cat. AB - A 13-year-old castrated domestic shorthair cat was examined because of fever, anorexia, and dermatologic lesions. Crusting, erythema, and well-demarcated purple discoloration of the foot pads and the tips of the pinnae, nose, and tail were seen. A white flocculent precipitate was detected in cooled serum. This precipitate dissolved upon rewarming, consistent with a cryoglobulin. Hypercalcemia, high alanine and aspartate aminotransferase activities, thrombocytopenia, and a monoclonal IgG gammopathy were found. Numerous hepatic nodules were detected by means of abdominal ultrasonography. Cytologic evaluation of fine-needle aspirates of the liver and spleen revealed numerous plasma cells, and evaluation of a bone marrow aspirate revealed plasmacytosis. A diagnosis of multiple myeloma and monoclonal IgG cryoglobulinemia was made, and the cat was euthanatized. PMID- 11019711 TI - Hypercalcemia following renal transplantation in a cat. AB - An 11-year-old 3.0-kg (6.6-lb) neutered male Persian was referred for renal transplantation. Serum total calcium concentration was slightly high prior to surgery, but the week after surgery, total and ionized calcium concentrations were extremely high, and a small mass was palpable on the right side of the trachea at the level of the thyroid and parathyroid glands. Exploratory surgery of the ventral aspect of the neck was performed, and a right external parathyroid mass was removed. One hour after surgery, the serum ionized calcium concentration was within reference limits, and the serum calcium concentration remained normal for the next 14 months without any specific treatment. The gross and histologic appearance of the mass, combined with the rapid decrease in serum calcium concentration following its removal, confirmed that the mass was a functional parathyroid adenoma. Although a common postoperative complication in people, hypercalcemia following renal transplantation appears to be a rare complication in cats. Surgery should be considered if the condition is a result of a parathyroid adenoma. PMID- 11019712 TI - Osteosarcoma in adjacent lumbar vertebrae in a dog. AB - An 8-year-old male Belgian Malinois was referred for evaluation of progressive caudal paresis of 2 to 3 weeks' duration. Radiography revealed a mottled appearance to the body of L4 and misshapen intervertebral foramen at L4-L5. Myelography revealed that the dye column terminated within the body of L4. Computed tomography revealed a soft tissue mass adjacent to or involving the spinal cord and L4, with complete destruction of a portion of the floor of the vertebral foramen. Small circular lesions were also noticed within the body of L3 and L5. A left-sided hemilaminectomy was performed. Histologic examination of a biopsy specimen revealed a high-grade sarcoma. Because of the poor prognosis, the dog was euthanatized. Necropsy examination revealed osteosarcoma, with lesions in L3 to L7, the sacrum, and the lungs. Metastatic lesions in adjacent bones have been termed skip metastases and the primary tumor is typically in long bones. Prognosis associated with skip metastases is similar to or even graver than that associated with pulmonary metastases. In the dog of the present report, the unusual finding of distinct foci of osteosarcoma within 5 adjacent lumbar vertebrae and the sacrum was consistent with skip metastases, potentially spread via the vertebral venous plexus. PMID- 11019713 TI - Lung lobe torsion in dogs: 22 cases (1981-1999). AB - OBJECTIVE: To identify breed disposition, postoperative complications, and outcome in dogs with lung lobe torsion. DESIGN: Retrospective study. ANIMALS: 22 client-owned dogs. PROCEDURE: Information on signalment; history; clinical findings; results of clinicopathologic testing, diagnostic imaging, and pleural fluid analysis; surgical treatment; intra- and postoperative complications; histologic findings; and outcome were obtained from medical records. RESULTS: All 22 dogs had pleural effusion; dyspnea was the most common reason for examination. Fifteen dogs were large deep-chested breeds; 5 were toy breeds. Afghan Hounds were overrepresented, compared with the hospital population. One dog was euthanatized without treatment; the remaining dogs underwent exploratory thoracotomy and lung lobectomy. Eleven dogs recovered from surgery without complications, but 3 of these later died of thoracic disease. Four dogs survived to discharge but had clinically important complications within 2 months, including chylothorax, mediastinal mesothelioma, gastric dilatation, and a second lung lobe torsion. Six dogs died or were euthanatized within 2 weeks after surgery because of acute respiratory distress syndrome, pneumonia, septic shock, pneumothorax, or chylothorax. Chylothorax was diagnosed in 8 of the 22 dogs, including 4 Afghan Hounds. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lung lobe torsion is rare in dogs and develops most frequently in large deep chested dogs, particularly Afghan Hounds. Other predisposing causes were not identified, but an association with chylothorax was evident, especially in Afghan Hounds. Prognosis for dogs with lung lobe torsion was fair to guarded. PMID- 11019715 TI - Hemithyroidectomy in a horse with confirmed hyperthyroidism. AB - A 23-year-old Quarter Horse gelding was admitted to the hospital because of cachexia and hyperactive behavior of 1 year's duration. At admission the horse was severely emaciated, tachycardic with a grade V/VI diastolic murmur, pyrexic, polydipsic, enophthalmic, and alopecic. The right lobe of the thyroid gland was noticeably larger than typical. The horse was also hyperexcitable and had a ravenous appetite. A presumptive diagnosis of hyperthyroidism was made on the basis of clinical signs and high plasma thyroid hormone concentrations. Confirmation of the diagnosis was made on the basis of results of a triiodothyronine-suppression test. Following endocrine testing, the affected portion of the thyroid gland was removed and identified histologically as an adenoma. Return or plasma thyroid hormone concentrations to reference range values and resolution of the clinical signs of disease following hemithyroidectomy provided further conformation of the diagnosis. On the basis of finding in this horse, it appears that horses with hyperthyroidism may be successfully treated by hemithyroidectomy. PMID- 11019714 TI - Serologic confirmation of Ehrlichia equi and Borrelia burgdorferi infections in horses from the northeastern United States. AB - OBJECTIVE: To determine whether horses living in tick-infested areas of northeastern United States with clinical signs of borreliosis or granulocytic ehrlichiosis had detectable serum antibodies to both Borrelia burgdorferi and Ehrlichia equi. DESIGN: Prospective study. ANIMALS: Serum samples from 51 clinically normal horses, 14 horses with clinical signs of borreliosis, and 17 horses with clinical signs of granulocytic ehrlichiosis. PROCEDURE: Serum B burgdorferi or E equi antibodies were measured by use of an ELISA, immunoblot analysis, or indirect fluorescent antibody (IFA) staining. RESULTS: Of the 82 serum samples tested, 37 (45.1%) and 13 (15.9%) had detectable antibodies to B burgdorferi or E equi, respectively. Test results indicated that 12 horses had been exposed to both agents, 11 of these horses had granulocytic ehrlichiosis. The ELISA regularly detected antibodies to the following recombinant protein (p) antigens of B burgdorferi: p29, p37, p39, and p41-G. The use of immunoblot analysis confirmed ELISA results by indicating antibody reactivities to antigens of whole-cell B burgdorferi having molecular masses of predominantly 31, 34, 37, 39, 41, 58, and 93 kd. CONCLUSIONS AND CLINICAL RELEVANCE: Horses living in areas where ticks (Ixodes scapularis) abound are sometimes exposed to multiple pathogens. Analyses for specific recombinant borrelial antibodies using an ELISA can help separate horses with borreliosis from those with granulocytic ehrlichiosis, even when antibodies to both etiologic agents are detected in serum samples. Analysis using immunoblots is sensitive, and along with ELISA or IFA procedures, is suitable for confirming a clinical diagnosis of each disease. PMID- 11019716 TI - Management of patients with non-ST-segment elevation acute coronary syndromes: insights from the PURSUIT trial. AB - The glycoprotein (GP) IIb-IIIa inhibitor eptifibatide (INTEGRILIN, COR Therapeutics, Inc., South San Francisco, California, and Key Pharmaceuticals, Inc., Kenilworth, New Jersey) is a novel and highly potent antithrombotic agent indicated for the management of patients with non-ST-segment elevation acute coronary syndromes (ACS) and those undergoing percutaneous coronary intervention. The approval of eptifibatide for non-ST-segment elevation ACS was based on the positive results of the Platelet Glycoprotein IIb/IIIa in Unstable Angina: Receptor Suppression Using Integrilin Therapy (PURSUIT) trial. With enrollment of almost 11,000 patients, not only is the PURSUIT trial the largest trial of a GP IIb-IIIa inhibitor to date, but it is also the largest clinical study ever conducted in patients with non-ST-segment elevation ACS. The key feature of the PURSUIT trial is that patient management closely resembled standard clinical practice, because decisions about the use and timing of invasive cardiac procedures were made by the individual physicians rather than being prespecified in the study protocol. Eptifibatide therapy was associated with a significant reduction in the incidence of the primary endpoint--a composite of death or myocardial infarction at 30 days (14.2 vs. 15.7% in the placebo group; p = 0.042). Of importance is the fact that the beneficial effect of eptifibatide was independent of the management strategy pursued during study drug infusion (invasive or conservative), and it was achieved with few major safety concerns. These findings demonstrate that the use of eptifibatide should be considered for all patients presenting with signs and symptoms of intermediate- to high-risk non ST-segment elevation ACS. PMID- 11019717 TI - Acute lung injury and acute respiratory distress syndrome. The clinical syndrome. AB - Acute lung injury (ALI) is a syndrome of severe acute respiratory failure defined by a constellation of clinical criteria. The exact incidence of ALI is not known, but it generally occurs in the setting of acute severe illness. The course of ALI after onset is quite variable, but outcome is associated with risk factor, age, and comorbidity. Survival from this syndrome has improved over time. Survivors often are impaired after hospital discharge but tend to improve over time; however, ALI does confer a significant additional burden on survivors with regard to pulmonary function and health-related quality of life. PMID- 11019718 TI - Radiographic findings in patients with acute respiratory distress syndrome. AB - Since its description in 1967, acute respiratory distress syndrome (ARDS) has become a widely recognized, if somewhat imperfectly understood, entity. This article reviews the imaging characteristics of ARDS as demonstrated on plain chest radiography, CT scan, radionuclide imaging, and MR imaging. The abnormalities displayed on these modalities are well understood even though there may be some dispute as to their relative importance in diagnosing and managing patients. PMID- 11019719 TI - Pulmonary pathology of acute respiratory distress syndrome. AB - Lung morphology in ARDS reflects the rapid evolution from interstitial and alveolar edema to end-stage fibrosis consequent to injury of the alveolocapillary unit. This morphologic progression, termed diffuse alveolar damage, has been subdivided into sequentially occurring exudative, proliferative, and fibrotic phases. Pulmonary lesions correlate with the phase of alveolar damage rather than its specific cause. The pathologic features are consistent with the effects of a host of injurious stimuli and the complex interaction of inflammatory mediators on alveolar epithelial and capillary endothelial cells. Although ARDS frequently culminates in "interstitial" fibrosis, the organization of intraluminal exudate dominates the histologic picture in the proliferative phase and establishes the framework for subsequent fibrous remodeling of the lung. Involvement of the pulmonary vasculature is an important aspect of ARDS, from the initial phase of edema to the terminal stage of intractable pulmonary hypertension. Vascular lesions include thrombotic, fibroproliferative, and obliterative changes that, like the parenchymal lesions, correlate with the temporal phase of DAD. Although ARDS is characterized by extensive bilateral lung involvement, alveolar damage can also affect the lung in a localized fashion. RAD is associated with the same clinical risk factors as DAD, suggesting that there is a spectrum in the extent of lung involvement and disease severity in patients at risk for ARDS. The factors that govern which patients will develop the fulminant syndrome are poorly understood. It must be re-emphasized that the lung is stereotyped in its response to injury and, consequently, descriptive, or even quantitative, studies of lung morphology can only provide clues regarding the initiating factors and pathogenetic mechanisms of ARDS. Progress in understanding the pathogenesis of ARDS and development of rational approaches to therapy will ultimately depend on careful clinical and experimental studies and the application of immunohistochemical and molecular biology techniques to unravel basic mechanisms of cellular injury and response. PMID- 11019720 TI - Mediators and mechanisms of acute lung injury. AB - Since last reviewed in this forum, there have been remarkable advances in our understanding of the acute inflammatory process and how it contributes to the development of ALI. As stated in the beginning of this article, it is not possible to even begin to review all the specific advances that have been made. Instead, the author has focused on concepts that have emerged and improved our ability to study the pathogenesis of ARDS. These include the recognition that patients at risk for and with ARDS represent a heterogeneous population, that mediators or markers of inflammation cannot be considered in isolation, that a balance between proinflammatory mediators and inflammatory modulators may be important, and that there are several genetic factors that could contribute to the susceptibility for the development of ARDS. Hopefully these concepts can be expanded and clarified so that the next review of this topic can report on successful therapeutic interventions for the prevention and the treatment of ARDS. PMID- 11019721 TI - Alveolar epithelial barrier. Role in lung fluid balance in clinical lung injury. AB - Several studies have established that transport of sodium from the air spaces to the lung interstitium is a primary mechanism driving alveolar fluid clearance, although further work is needed to determine the role of chloride in vectorial fluid transport across the alveolar epithelium. Although there are significant differences among species in the basal rates of sodium and fluid transport, the basic mechanism seems to depend on sodium uptake by channels on the apical membrane of alveolar type II cells, followed by extrusion of sodium on the basolateral surface by Na,K-ATPase. This process can be upregulated by several catecholamine-dependent and independent mechanisms. The identification of water channels expressed in lung, together with the high water permeabilities, suggest a potential role for channel-mediated water movement between the air space and capillary compartments, although definitive evidence will depend on the results of transgenic mouse knock-out studies. The application of this new knowledge regarding salt and water transport in alveolar epithelium in relation to pathologic conditions has been successful in clinically relevant experimental studies, as well as in a few clinical studies. The studies of exogenous and endogenous catecholamine regulation of alveolar fluid clearance are a good example of how new insights into the basic mechanisms of alveolar sodium and fluid transport can be translated to clinically relevant experimental studies. Exogenous catecholamines can increase the rate of alveolar fluid clearance in several species, including the human lung, and it is also apparent that release of endogenous catecholamines can upregulate alveolar fluid clearance in animals with septic or hypovolemic shock. It is possible that therapy with beta adrenergic agonists might be useful to accelerate the resolution of alveolar edema in some patients. In some patients, the extent of injury to the alveolar epithelial barrier may be too severe for beta-adrenergic agonists to enhance the resolution of alveolar edema, although some experimental studies indicate that alveolar fluid clearance can be augmented in the presence of moderately severe lung injury. A longer-term upregulation of alveolar epithelial fluid transport might be achieved by strategies that accelerate the proliferation of alveolar type II cells repopulating the injured epithelium in clinical lung injury. More clinical research is needed to evaluate the strategies that can upregulate alveolar epithelial fluid transport with both short-term therapy (i.e., beta agonists) and more sustained, longer-term effects of epithelial mitogens such as keratinocyte growth factor. These approaches may be useful in reducing mortality in the acute respiratory distress syndrome. PMID- 11019722 TI - Mechanical ventilation in acute lung injury and acute respiratory distress syndrome. AB - Mechanical ventilation provides life-sustaining support for most patients with acute lung injury and acute respiratory distress syndrome; however, traditional approaches to mechanical ventilation may cause ventilator-associated lung injury, which could exacerbate or perpetuate respiratory failure caused initially by conditions such as pneumonia, sepsis, and trauma. This article reviews the theory, laboratory data, and results of recent clinical trials that suggest that modified ventilator strategies can reduce ventilator-associated lung injury and improve clinical outcomes. PMID- 11019723 TI - Prone ventilation. AB - Considerable clinical experience confirms that oxygenation can be improved in many patients with ARDS by employing prone ventilation. The improvement occurs because, in the prone position, the lung fits into the thorax such that lung distention is more uniform and compressive forces extant in the supine position, which serve to cause dorsal airspace collapse, are reduced. Whether these changes translate into improved clinical outcomes has yet to be determined, but prone ventilation has the potential of reducing oxygen toxicity and limiting ventilator induced lung injury. PMID- 11019724 TI - Inhaled nitric oxide and acute lung injury. AB - The nitric oxide (NO) field has been one of the most exciting scientific ventures over the past 10 years. Among the researches developed, the use of inhalation of NO gas allowed us to propose this therapy in lung diseases with promising results. Because of its property as a "selective" pulmonary vasodilator and because of its apparent clinical safety, inhaled NO has been proposed in acute lung injury (ALI) to improve severe hypoxemia. In this situation, the abnormal ventilation-perfusion ratio is improved by inhaled NO, limiting arterial hypoxia. The major clinical trials performed in adults, however, have failed to show any benefit on mortality and on mechanical ventilation requirements. Inhaled NO has been shown as an efficient therapy in pediatric ALI, probably because of a lower comorbidity. Because of the inhaled NO uptake by the lung, the extra vascular lung effects might be in the future the most important development in relation with platelet anti-agregant and anti-inflammatory properties. PMID- 11019725 TI - Surfactant replacement therapy. AB - Dysfunction of the surfactant system of the lung in the setting of acute lung injury (ALI) is likely to contribute to the pathophysiology of that syndrome. Multiple mechanisms, including injury to alveolar type II cells and inhibition by plasma proteins contribute to this loss of function. Similar injury occurs in animal models of acute lung injury and, in that setting, treatment with exogenous surfactant causes marked improvement in gas exchange. Clinical studies of surfactant treatment of ALI suggest benefit, and definitive phase III trials are now in progress. PMID- 11019726 TI - Partial liquid ventilation for acute respiratory distress syndrome. AB - PLV represents an intriguing alternative paradigm in the approach to the patient with ALI. Within the past decade, substantial information has become available regarding this technique. Clearly, PLV is feasible in patients with ALI and ARDS, and it appears to be safe with respect to short-term effects on hemodynamics and lung physiology, as well as long-term toxicity (although further research in this area is warranted). Although PLV has not yet been proven to be superior to traditional mechanical ventilation for patients with ALI or ARDS, PLV possesses an intriguing combination of physical, physiologic, and biologic effects: "Liquid PEEP" effect--e.g., more effective recruitment of dependent lung zones than achieved by gas ventilation Anti-inflammatory effects Lavage of alveolar debris Mitigation of ventilator-induced lung injury Direct anti-inflammatory effects- e.g., decreased macrophage release of proinflammatory cytokines, etc. Prevention of nosocomial pneumonia Combination with other modalities--e.g., exogenous surfactant replacement, inhaled NO, prone position Enhanced delivery of drugs or gene vectors into the lung. The results of ongoing and future clinical trials will be necessary to establish whether PLV improves clinical outcomes in patients with ALI or ARDS, or specific subgroups of such patients. Significant work also remains to be done to define the optimum dose level of PLV and the most appropriate ventilatory strategies. PMID- 11019727 TI - Extracorporeal life support in the management of severe respiratory failure. AB - ECLS is a safe and effective means to keep patients alive during severe respiratory failure that would otherwise be fatal. In addition to direct and indirect treatment of the lungs during ECLS, the technique allows days of time for study and treatment of other conditions and other organ failure. The technique has been refined in newborn infants and children, in whom survival rates are high and the technology is proven by prospective randomized trials. ECLS is usually applied to adults with respiratory failure when the mortality risk is over 80%. With these indications, the survival rate in experienced centers is 50% to 60%. A new prospective, randomized trial of ECLS in adult patients is underway in the United Kingdom. In the meantime, intensivists who are charged with the management of moribund ARDS patients who fail to respond to other methods of therapy should consider the risks versus the benefits of transferring such patients to an ECLS center. PMID- 11019728 TI - Acute respiratory distress syndrome. Potential pharmacologic interventions. AB - Remarkable progress has been made in the past 10 years with regard to understanding the interplay of potent physiologic mediators in patients with acute lung injury. Because there are so many mediators and the interaction of these agents is complex, true insight into the process has been slow in coming. Clinical studies in ARDS, as well as sepsis, the leading cause of ARDS, have increased in number, size, and quality over this same period. Although none of these studies has produced an accepted new therapy for ARDS, each has laid the groundwork for more efficient and more elegant studies of the problem. The stage is now set for the real advances to be brought forward and put to rigorous, efficient clinical testing. PMID- 11019729 TI - Mechanisms of repair and remodeling following acute lung injury. AB - At present, we largely lack the ability to correlate the clinical course of ARDS patients with potential factors involved in the biochemical and cellular basis of lung repair. This requires very large patient databases with measurement of many biochemical parameters. Important mechanistic determinants during the repair phase can be sought by correlation with late outcomes, but a large-scale cooperative effort among multiple centers with sharing of follow-up data and patient specimens is essential. We also lack detailed human histologic material from many phases of ARDS and, particularly, know little of the long-term morphologic impact of ARDS in survivors. Establishment of a national registry that follows ARDS survivors and that would seek their cooperation in advance in obtaining autopsy specimens when they die of other causes would be very valuable. Correlating the pathology with their pulmonary function during recovery would give important insights into the reasons for the different patterns of abnormal pulmonary functions. The factors that determine the success of repair are of critical importance in testing new ARDS treatment strategies. Would accelerating the resolution of alveolar edema alter the course of subsequent fibrosis and inflammation? Does surfactant replacement therapy--a costly proposition in adults with ARDS--lead to better long-term outcomes in survivors? How much should we worry about the use of high levels of oxygen for support of arterial partial pressure of oxygen? Is it better to accept hyperoxia to avoid pressure or volume trauma induced by mechanical ventilation with higher minute ventilations? These major management issues all may affect the success of the late repair and recovery process. Intervention trials need to examine the long-term physiologic and functional outcomes. PMID- 11019730 TI - The adolescent preparticipation physical examination. Is it helpful? AB - The preparticipation examination (PPE) is a widely practiced method of screening athletes for medical limitations before sports participation. Although the PPE is widely implemented, there is little consistency regarding content or method in providing this important service. This lack of uniformity undermines the overall effectiveness of the PPE. The history, efficiency, and nature of athletic screening in adolescents are discussed in this article, and specific suggestions are offered on how the examination might be improved in the future. As sports participation among teens continues to change--in number of participants and in athlete-demographics--and as the field of sports medicine expands equally rapidly, the adolescent PPE, the commonest interaction between adolescent patients and the medical system, should be updated. PMID- 11019731 TI - Strength training for children and adolescents. AB - The potential benefits of youth strength training extend beyond an increase in muscular strength and may include favorable changes in selected health- and fitness-related measures. If appropriate training guidelines are followed, regular participation in a youth strength-training program has the potential to increase bone mineral density, improve motor performance skills, enhance sports performance, and better prepare our young athletes for the demands of practice and competition. Despite earlier concerns regarding the safety and efficacy of youth strength training, current public health objectives now aim to increase the number of boys and girls age 6 and older who regularly participate in physical activities that enhance and maintain muscular fitness. Parents, teachers, coaches, and healthcare providers should realize that youth strength training is a specialized method of conditioning that can offer enormous benefit but at the same time can result in serious injury if established guidelines are not followed. With qualified instruction, competent supervision, and an appropriate progression of the volume and intensity of training, children and adolescents cannot only learn advanced strength training exercises but can feel good about their performances, and have fun. Additional clinical trails involving children and adolescents are needed to further explore the acute and chronic effects of strength training on a variety of anatomical, physiological, and psychological parameters. PMID- 11019732 TI - Acute knee injuries. AB - Skeletally immature athletes do get major knee injuries. Hemarthrosis is associated with peripheral meniscal tears, anterior cruciate ligament ruptures, tibial tubercle avulsion injuries, and patellar/femoral osteochondral fractures and cannot be ignored. The primary diagnostic tool for patients with a knee injury is a clinical examination by a physician well trained in knee evaluation. MR imaging has significant limitations in this age group. The algorithm for anterior cruciate ligament injury treatment must take into account the patient's physiologic maturity, not chronological age. PMID- 11019733 TI - Lower limb injuries in children in sports. AB - This article presents an overview of sports-related injuries of the lower limb in children, with emphasis on the management. The special injury-related conditions of childhood, epidemiology, and the particular pattern of injuries are discussed. The increased participation of children in sports will continue. Permanent damage is a risk, and, as such, prevention should be the most important management in this age group. For example, stretching exercises should be performed with "warm" muscles. Excessive weight training has an unacceptable risk of injury. In endurance sports, the "10 percent rule," which consists of increasing activity by 10% each week, probably could be applied to prevent overuse injuries. A multifactorial approach may be helpful. The rationale for high performance, competitive sports for children is doubtful. The optimal levels of safe training will remain changeable and not predictable. It should be the responsibility of parents, coaches, and healthcare professionals to try and minimize the potential for injury and disability, and allow children to enjoy the benefits of sports. PMID- 11019734 TI - Back injuries in the young athlete. AB - The diagnosis of back pain in the young athlete should be specific and not attributed to nonspecific, mechanical causes. Risk factor identification and intervention are required. Treatment is then initiated in a specific pattern, addressing flexibility and muscular imbalances. Bracing is often used to allow healing of growth tissue. The lumbosacral orthosis may be molded in a lordotic posture to unload the disc or antilordotic posture to relieve the posterior column; however, customizing the lordosis to the individual biomechanics may be required. Spinal stabilization is initiated with therapy for strengthening isolated weaknesses and progressing to coactivation and proprioceptive techniques, such as the balance ball. Returning to competition is preceded with sport-specific training. PMID- 11019735 TI - Shoulder injuries in the childhood athlete. AB - Shoulder dysfunction in the young athlete usually is manifested differently than the adult counterpart. The physiology and biomechanics of a growing child and adolescents result in different injury patterns that require different and thoughtful approaches to diagnosis and treatment. Most of these conditions are served well by nonsurgical treatment modalities. Judicious use of surgical interventions, however, can significantly improve patient outcome and return them to their sport of interest. Postoperative rehabilitation, and proper training techniques are essential to ensure continued participation of the athlete. PMID- 11019736 TI - Head and neck injuries in young athletes. AB - Careful study of the pathophysiology and epidemiology of sports-related spine injuries brings to light many common features. The incidence increases as the sport becomes increasingly violent and aggressive. Poor conditioning and lack of knowledge of the proper techniques of the sport put the athlete at significant risk for head and spine injury. Improper helmet fit and the use of the head as an offensive weapon also are common features of injury. Although recognition of these features has resulted in a dramatic reduction in catastrophic neurological injury, the athlete remains at risk for less severe head and spine injury, and concussion remains at epidemic proportions at high school, university, and professional levels. It is hoped that careful recognition of the signs of concussion and knowledge of return-to-play criteria will prevent catastrophic complications from minor head injuries, although the long-term effects of multiple concussions on cognition may be problematic. PMID- 11019737 TI - The young dancer. AB - The injuries that are prevalent in and unique to dancers have their origins inextricably linked to faulty technique or poor biomechanics, combined with other risk factors. It is this combination of factors that must be addressed when considering diagnosis, treatment, and rehabilitation. For the dancer to return to full activity with minimal risk of recurrent injury, neuromuscular re-education is mandatory. This process best entails using a team approach, accessing the resources of healthcare professionals and those who train the dancer on a daily basis. In some cases, parents also may need to be involved. Communication, interaction, and mutual understanding among these groups will assist the dancer in regaining and maintaining health. PMID- 11019738 TI - The young skater. AB - Figure skating is one of the best lifetime sports. Participants may skate alone or with a group, recreationally or competitively, at all ages and at all skill levels. As a sport that improves physical fitness, skating builds strength, flexibility, and cardiopulmonary endurance. As a weight-bearing or high-impact activity, it is one of the best sports for increasing bone mineral density and perhaps for preventing osteoporosis. At least half of all competitive figure skating injuries appear preventable. Boots should be as flexible as the skater can control and should be carefully fitted. Normal flexibility of lower extremity muscles, especially in the growing athlete, markedly decreases the incidence of overuse symptoms of the knee. Core body strength is critical for controlling high impact jump landings and decreasing injuries to the spine, pelvis, and hip girdle. Adequate shoulder strength is required for pair skaters and ice dancers. A well-designed off-ice training program decreases injury incidence and enhances performance. Assisting these dedicated athletes is truly rewarding as they train and compete at their highest potential, with passion for their sport. PMID- 11019739 TI - The young gymnast. AB - Gymnastics training develops strength, flexibility, concentration, balance, grace, and speed in young athletes. In terms of hours per week and intensity of practice, the dedication to training at a young age, is unparalleled in most other youth sports. With this dedication comes the risk of injury to the immature musculoskeletal system, and it is our duty to ensure that these risks are minimized. Through adequate safety equipment, supervision, and spotting of difficult techniques, a number of acute injuries may be prevented. Maintenance of balanced flexibility and strength, modification of training to limit pain, and taping or splinting of wrists and ankles may reduce the risk of overuse injuries. Physician who deal with young gymnasts must have an understanding of the inherent risks of the sport to provide prompt diagnoses and appropriate management of injuries. The gymnasium can be a healthy environment as long as the health and safety of the child takes precedence over the success of the gymnast. PMID- 11019740 TI - Musculoskeletal injuries in the young tennis player. AB - Tennis is becoming increasingly popular, especially with young athletes. Despite recent advances in epidemiologic research of tennis injuries, there still is a need for more injury research in all of the racquet sports. The data that does exist show that the young athlete is susceptible to injury in these different sports. Injury patterns in the skeletally immature racquet sports athlete are becoming apparent. Although most of the sports result in similar injury patterns, such as a predominance of lower extremity injury, there are differences. It appears that the physical demands of the sport are becoming more clearly documented, and the adaptive response to these demands is becoming understood. The adaptive response reveals a common origin for many of the injuries in the different sports. This is related most often to repetitive microtrauma with resultant loss in flexibility and strength. The sports medicine practitioner must understand these differences, know the demands, do serial musculoskeletal evaluations for maladaptations, and adhere to a periodized prehabilitation program of preventative exercises to maximize performance and minimize injury risk. PMID- 11019741 TI - The young athlete with physical challenges. AB - This article is intentionally broad in scope, as a result of a collaboration from the fields of primary care sports medicine, orthopedic surgery, and kinesiology. What has been borne out in the process is a true appreciation of the benefits of a multidisciplinary approach toward providing care for the young athlete with a physical disability. To name a few, joint involvement of parents, coaches, trainers, physical therapists, orthotists, prosthetists, wheelchair engineers, neurologists, physiatrists, nutritionists and most importantly, the athletes themselves, should be further encouraged because each discipline provides a unique perspective in the identification and management of health-related issues. It is the intent of this article to provide readers with at least some new insight that they can carry into their future practice. PMID- 11019742 TI - The prevention of sports injuries in children. AB - With the explosion of the number of youth participating in sports activities has come an epidemic of injuries. Myriad factors make their prevention a great challenge for society. This article presents an overview of statistics, factors contributing to sports injuries, a review of significant prevention efforts, barriers to prevention, and recommendations for the future. PMID- 11019743 TI - Modeling and measuring the personality disorders. AB - Livesley, Jang, and Vernon (1998. Archives of General Psychiatry, 55, 941-948) reported that personality disorders (PDs) are quantitatively extreme expressions of normal personality functioning. A similarly designed study attempts to replicate those findings for both self- and observer-rated reports of patients judged clinically to have a PD. Analyses of data sets generated by 758 self reports (SRs) and 515 reports from corroborative witnesses (CWs) refined the set of 266 descriptors to 142 items assessing 30 constructs. Intercorrelation of the constructs revealed considerable interdependence. Principal components analyses identified four factors, consistent across the SR and CW databases, and consistent with the Livesley et al. (1998) analyses where they were labeled Emotional Dysregulation. Dissocial, Inhibition, and Compulsivity. Replication and extension of findings to CW-rated data offer additional support for the argument that higher-order PD traits strongly resemble normal personality dimensions. PMID- 11019744 TI - The SCL-90-R as a screening instrument for severe personality disturbance among outpatients with mood and anxiety disorders. AB - Our objective was to examine whether the SCL-90-R, a widely used self-report measure of distress and psychopathology, could screen for personality disorders in general and for severe personality disturbance (SPD) in particular, at the time when patients seek treatment for a state (mood or anxiety) disorder. The SCL 90-R was administered to 112 consecutive outpatients with various mood and anxiety disorders. The personality severity index (PSI) score, defined as the mean value of the scores on the SCL-90-R subscales of interpersonal sensitivity, hostility, and paranoid ideation, was compared with the current symptom index (CSI) score, defined as the mean value of the scores on the remaining six SCL-90 R subscales. A positive screen was considered if PSI > CSI. SPD was defined as the presence of any DSM-III-R Cluster A and/or Cluster B personality disorder. The accuracy of the screen was verified by means of the Structured Clinical Interview for DSM-III-R Personality Disorders. The sensitivity of the PSI > CSI criterion to screen for SPD was 89.4%, while its sensitivity to screen for any DSM-III-R personality disorder was 72.9%. The SCL-90-R subscales that contributed the most to the screening discriminability of this SCL-90-R-derived screening measure were hostility, paranoid ideation, somatization, and obsessive-compulsive behavior. The SCL-90-R may be used to screen for SPD in routine work with outpatients with mood and anxiety disorders, but the results of the screening need to be verified because of the possibility of false negatives and false positives, although that possibility is apparently low. These findings may have important prognostic and treatment implications. PMID- 11019745 TI - A family study of outpatients with borderline personality disorder and no history of mood disorder. AB - While several studies have examined psychiatric disorders in the relatives of individuals with borderline personality disorder, many of these studies have not employed a family study methodology and suffer from other methodological shortcomings. Thus, the conclusions from family data addressing the validity of borderline personality disorder, its relation to other conditions, and its distinction from mood disorders, continue to be debated. The present investigation employed a family study design with direct interviews with relatives, structured diagnostic interviews with both probands and relatives, and blind assessment of relatives. Rates of psychiatric disorders were examined in 563 relatives of outpatients with mood disorders (n = 119), 54 relatives of outpatients with borderline personality disorder and no history of mood disorder (n = 11), and 229 relatives of never psychiatrically ill controls (n = 45). Results indicate increased rates of mood disorders and personality disorders in the relatives of borderline probands compared with never psychiatrically ill controls. Familial aggregation of psychiatric disorders was generally similar for borderline personality and the mood disorder comparison group. The results suggest there may be common etiological factors between borderline personality disorder and mood disorders. PMID- 11019746 TI - Gender bias in self-report personality disorder inventories. AB - There has been considerable controversy and research regarding gender biases in the diagnosis of personality disorders, but few studies have explored whether personality disorder self-report inventories might contain gender biases. The current study investigated whether items from three commonly used inventories evidence a potential for gender bias. Subjects were from outpatient mental health clinics. Items were considered gender biased if they exhibited gender differences and failed to correlate with or, more importantly, correlated negatively with dysfunction. Thirty-eight items evidenced potential bias, the majority of which were from Narcissistic scales. The implications of the results for the clinical assessment of purportedly maladaptive personality traits and for the construction of personality disorder scales are discussed. PMID- 11019747 TI - Negative childhood accounts, sensitivity, and pessimism: a study of avoidant personality disorder features in college students. AB - Theory suggests that individuals with avoidant personality disorder (APD) may have experienced rejection or isolation during childhood, that they may be temperamentally hypersensitive, and that they may hold pessimistic expectancies. In a sample of 127 undergraduates, negative childhood memories, sensory processing sensitivity, and pessimism were assessed. To measure childhood memories, participants wrote open-ended narratives that were evaluated for valence by independent raters. To measure APD features, participants rated the degree of which verbatim DSM-IV criteria were descriptive of themselves. Negative childhood accounts, self-reports of sensory-processing sensitivity, and pessimism were correlated with DSM-IV APD features, even after controlling for the influence of present negative mood. Interactions suggested that pessimism was more strongly related to APD features among participants who were either highly sensitive or recalled adverse childhood experiences (e.g., isolation, rejection, conflict). Implications for theory and intervention are discussed. PMID- 11019748 TI - The cross-cultural generalizability of Axis-II constructs: an evaluation of two personality disorder assessment instruments in the People's Republic of China. AB - We examined the reliability, cross-instrument validity, and factor structure of Chinese adaptations of the Personality Diagnostic Questionnaire (PDQ-4+; N = 1,926) and Personality Disorders Interview (PDI-IV; N = 525) in psychiatric patients. Comparisons with data from Western countries suggest that the psychometric properties of these two instruments are comparable across cultures. Low to modest agreement between the PDQ-4+ and PDI-IV was observed for both dimensional and categorical personality disorder evaluations. When the PDI-IV was used as the diagnostic standard, the PDQ-4+ showed higher sensitivity than specificity, and higher negative predictive power than positive predictive power. Factor analyses of both instruments replicated the four-factor structure O'Connor and Dyce (1998) found in Western samples. Results suggested that conceptions and measures of DSM-IV personality disorders are cross-culturally generalizable to Chinese psychiatric populations. PMID- 11019749 TI - Biparental failure in the childhood experiences of borderline patients. AB - The purpose of this study was to assess the role of biparental abuse and neglect in the development of borderline personality disorder (BPD). A semistructured research interview was used to blindly assess the childhood experiences of biparental abuse and neglect reported by 358 borderline inpatients and 109 axis II controls. Eighty-four percent of borderline patients reported having experienced some type of biparental abuse or neglect before the age of 18; 55% reported a childhood history of biparental abuse; 77% reported a childhood history of biparental neglect. These experiences were also reported by a substantial percentage of Axis II controls (biparental abuse or neglect [61%], biparental abuse [31%], and biparental neglect [55%]). However, borderline patients were significantly more likely than axis II controls to report having been verbally, emotionally, and physically but not sexually abused by caretakers of both sexes. They were also significantly more likely than controls to report having caretakers of both sexes deny the validity of their thoughts and feelings, fail to provide them with needed protection, neglect their physical care, withdraw from them emotionally, and treat them inconsistently. It was also found that female borderlines who reported a previous history of neglect by a female caretaker and abuse by a male caretaker were at significantly higher risk for having been sexually abused by a noncaretaker. Taken together, the results of this study suggest that biparental failure may be a significant factor in the etiology of BPD. They also suggest that biparental failure may significantly increase a preborderline girl's risk of being sexually abused by someone other than her parents. PMID- 11019750 TI - Major depression, double depression and personality disorders. AB - Personality disorders are much more common among depressive patients than among normal people. Until now, little research has been conducted into the prevalence of personality disorders among patients with both major depression and dysthymia (double depression). The subject of this study is whether depressive patients with dysthymia have more personality disorders than those with no dysthymia. The Vragenlijst voor Kenmerken van de Persoonlijkheid (a Dutch self-report based on the International Personality Disorder Examination) was completed for 211 outpatients with major depression. Approximately 60% of the patients suffer from one or more personality disorders. Depressive patients with dysthymia differ little from the patients without dysthymia, but patients with dysthymia have more cluster A disorders and are more avoidant. Depressive patients without dysthymia do not differ from the patients with dysthymia in terms of symptoms. Depressive patients with personality disorders have significantly more symptoms than the patients without these disorders. There is no interaction between dysthymia and personality disorder. PMID- 11019752 TI - Six lessons in medical nutrition education. PMID- 11019751 TI - The relationship between five-factor personality measurements and ICD-10 personality disorder dimensions: results from a sample of 229 subjects. AB - This article examines the relationship between the five-factor model (FFM) and dimensional ICD-10 personality disorders. In a follow-up study of a child and adolescent psychiatric cohort, former patients and controls were assessed with NEO-FFI and the IPDE interview (CD-10 personality disorder). Full data were available for 229 subjects (149 former patients, 80 controls). Multiple regression analysis showed that the five factors of the FFM as independent variables explained between 5% (schizoid personality disorder) and 32% (anxious personality disorder) of the variance of ICD-10 dimensional personality disorder scores. For the two types of emotionally unstable personality disorder dimension (impulsive and borderline), for anxious (avoidant) personality disorder dimension and for the total score of any personality disorder dimension, FFM explained between 17% and 32% of the variance with almost identical results for the former patient group and the control group. High neuroticism was a feature of paranoid, emotionally unstable, histrionic, anankastic, anxious (avoidant), and dependent personality disorder dimensions, whereas low agreeableness was found in dissocial, emotionally unstable and histrionic personality disorder dimensions. Low extraversion was found in schizoid, anxious (avoidant) and dependent personality disorder dimensions, whereas histrionic PD dimension correlated with high extraversion. We find that the FFM is valuable for the further understanding not only of DSM-IV but also of ICD-10 personality disorder dimensions. The differences between ICD-10 and DSM-IV in this respect seem to be small. PMID- 11019753 TI - Curricular changes, challenges, and opportunities in medical nutrition education. PMID- 11019754 TI - Nutrition-related education, research training, and career development opportunities at the National Cancer Institute. PMID- 11019755 TI - Nutrition and cancer education: ten years of progress. AB - The Nutrition Education and Research Program at the University of Nevada School of Medicine was awarded two separate NIH/NCI R25 cancer education grants over a ten-year period. With this support, a four-year longitudinal nutrition curriculum was implemented, including the required 20-hour freshman Medical Nutrition Course, junior and senior nutrition electives, and a senior assignment in nutrition and cancer during the rural rotation with faculty preceptors. Funding has also supported nutrition integration into the basic science courses, patient care courses, and specialty clerkships. A unique nutrition fellowship for medical students who specialize in nutrition during their four years of training and graduate with special Qualifications in Nutrition (SQIN) has also been instituted. The curriculum reflects a longitudinal, interdisciplinary, but flexible, integration of nutrition into an already crowded medical school education. PMID- 11019756 TI - An integrated nutrition curriculum in medical education. AB - R25 grant support from the NIH/NCI enabled the University of Arizona to assess nutrition education, develop and evaluate specific course content, and move toward comprehensive prevention-based nutrition education in 1991-1997. Hours of nutrition education increased to 115% over baseline, and students indicated greater satisfaction with the amount of nutrition instruction they received. Especially valuable was a course in prenatal and infant nutrition that paired each student with a pregnant patient. After the grant support ended, nutrition began to be crowded out of the curriculum by other, more traditional, topics, but a 57% gain over baseline was sustained. External support for nutrition education is urgently needed. PMID- 11019757 TI - An interdisciplinary training program in nutrition sciences and cancer. AB - To address the shortage of health care professionals trained in the nutritional aspects of cancer prevention, the University of Alabama at Birmingham in 1988 initiated the Cancer Prevention and Control Training Program (CPCTP), with R25 grant support from the NIH/NCI. The CPCTP has enrolled 11 predoctoral and 12 postdoctoral trainees, of whom 18 have completed training and five remain in the program. The curriculum and other program elements are described, and the trainees' academic achievements and ultimate careers are reported. The CPCTP has become a significant resource for training cancer nutrition professionals. PMID- 11019758 TI - A mini-fellowship in clinical nutrition for primary care physicians. AB - When the Regional Nutrition Center's (RNC's) mini-fellowship was being developed, physicians teaching in medical schools and residency programs had little formal training in nutrition, so it was difficult to identify faculty to serve as role models and clinical preceptors. Using funds provided by an NIH/NCI R-25 grant, the mini-fellowship in clinical nutrition was developed as a model to meet the nutrition education needs of senior residents and junior faculty in primary care disciplines. It provided a brief but broad exposure to clinical nutrition topics, with a focus on issues relevant to the practice of primary care and teaching skills. In four weeks, fellows completed didactic course work, a clinical preceptorship, and a teaching project. Tracking of the 55 physicians who completed the program by questionnaires has shown significant improvement in their nutrition-related patient care activities. PMID- 11019759 TI - Nutrition and cancer: a curriculum and short student research experiences. AB - In 1988, Louisiana State University Health Sciences Center (LSUHSC) received an NCI/NIH R25 grant to strengthen nutrition-cancer education in the medical school and to provide short cancer-related research experiences for students. Support for the nutrition-cancer curriculum component of the grant was limited to the first five years, while renewal of the research component was possible on a competitive basis. After the initial five-year award ended, the administration of LSUHSC assumed the support of the nutrition-cancer curriculum. This curriculum, modified over time, continues to be taught in each year of medical school and culminates with a required clinical nutrition course in the fourth year. The teaching of nutrition continues to be very successful and is well received by students and faculty. The research component, renewed twice, has achieved its goals over the same period. As a consequence of the initial award and the commitment of the administration, nutrition education at LSUHSC remains strong and is well established. PMID- 11019760 TI - The development of "Nutrition in Medicine" interactive CD-ROM programs for medical nutrition education. AB - In 1995 and in 1998 the University of North Carolina at Chapel Hill received R25 grant support to create interactive CD-ROMs for teaching nutrition and nutritional biochemistry to medical students, the Nutrition In Medicine (NIM) series. Seven of the proposed ten titles have been created. Three series (Disease, Lifecycle, and Special Topics in Nutrition) teach nutrition concepts using computer-aided instruction (CAI) with emphasis on interactive learning. Patient cases with television-quality interactive videos allow students to apply nutrition knowledge to clinical problems. Pop quizzes, text-based interactions, and multiple-choice examinations help the student self-evaluate progress via immediate feedback. Educators using the programs get instructional support and updates through a dedicated Web site, printed material, telephone support, e mail, and CD-ROM-based computer programs. Implementation at medical and osteopathic schools is continuously surveyed through questionnaires and follow-up telephone interviews. By 1999, 120 of 137 eligible U.S. medical schools owned copies of NIM CAI programs, of which 76 indicated that they were currently implementing the programs. PMID- 11019761 TI - Changes in nutrition knowledge among first- and second-year medical students following implementation of an integrated nutrition curriculum. AB - Existing studies suggest that many of the objectives called for by the National Academy of Sciences in its 1985 report are not being addressed in medical student education. Members of the Nutrition Curriculum Working Group at the UCLA School of Medicine prepared a list of proficiencies in nutrition and developed a coordinated, vertically integrated, two-year nutrition education curriculum to address those objectives. To assess the impact of the curriculum on students' knowledge, the authors tested a cohort of students at repeated intervals over a two-year period using a Nutrition Progress Survey drawn from the Nutrition Test Item Bank developed at the University of Alabama. There was a significant increase in knowledge over the three administrations of the Progress Survey as measured by total score. This was largely accounted for by an increase in the number of correct responses, which rose from 39% at pre-test to 62% at delayed follow-up. In spite of a significant increase in nutrition knowledge, the low percentage correct indicates that nutritional content should be threaded throughout the core clerkship curriculum as well. PMID- 11019762 TI - Nutrition and cancer education at the University of Colorado Health Sciences Center. AB - BACKGROUND: The status of nutrition education at the University of Colorado Health Sciences Center since the 1980s is discussed. METHODS: A needs assessment with emphasis on nutrition education activities in the medical school curriculum led to the successful application for a NCI R25 Cancer Education Grant in 1988. RESULTS: A nutrition elective, Nutrition and Cancer, for health professions students and a section of the Biochemistry course, Principles of Nutrition, were generated. These have persisted to the present despite the scheduled expiration of the R25 grant. CONCLUSION: Topic-directed NCI Cancer Education Grants can stimulate and facilitate successful approaches to defined cancer education needs. The resulting activities can, and should, persist after the expiration of such grants. PMID- 11019763 TI - Food avoidance by women at risk for recurrence of breast cancer. AB - BACKGROUND: This cross-sectional study examined self-reported restriction of foods by middle-aged and older women at risk for breast cancer recurrence, who were participating in an intervention study, and by women without breast cancer. METHODS: Data examined were height, weight, dietary intake, and avoidance of specific food types in women with breast cancer (n = 56) and a comparison group of women without breast cancer (n = 51). Data were analyzed with t-tests and chi square analysis. RESULTS: The broad food categories of meat, dairy foods, and sweets were restricted by women both with (53%, 74%, and 62%, respectively) and without breast cancer (47%, 63%, and 63%, respectively). Significantly more women with breast cancer avoided cakes, butter, and milk, compared with women without breast cancer (p < 0.05). Women in both groups attained a mean consumption of >75% of the Recommended Dietary Allowance for many nutrients; however, mean consumption for calcium was <75% of the RDA for the two groups. CONCLUSIONS: Analysis of food avoidance by women with breast cancer who participate in diet intervention studies and by women without breast cancer may provide information about adherence to dietary guidelines and adequate nutrient consumption among these populations. PMID- 11019764 TI - Dietary change through African American churches: baseline results and program description of the eat for life trial. AB - BACKGROUND: Eat for Life, a multicomponent intervention to increase fruit and vegetable (F & V) consumption among African Americans, is delivered through African American churches. METHODS: Fourteen churches were randomly assigned to one of three treatment conditions: 1) comparison; 2) culturally-sensitive multicomponent intervention with one phone call; and 3) culturally-sensitive multicomponent intervention with four phone calls. The intervention included an 18-minute video, a project cookbook, printed health education materials, and several "cues" imprinted with the project logo and a 5 A Day message. A key element of the telephone intervention was the use of motivational interviewing, a counseling technique originally developed for addictive behaviors. Major outcomes for the trial included total F & V intake, assessed by food-frequency questionnaires (FFQs) and 24-hour recalls, and serum carotenoids. Psychosocial variables assessed included outcome expectations, barriers to F & V intake, preference for meat meals, neophobia, social support to eat more F & V, self efficacy to eat more F & V, and nutrition knowledge. RESULTS: Baseline mean F & V intakes across the three FFQs ranged from 3.45 to 4.28 servings per day. Intake based on a single 24-hour recall was 3.0 servings. Variables positively correlated with F & V intake included self-efficacy, outcome expectations, and a belief that F & V contain vitamins. Factors negatively correlated with intake include perceived barriers, meat preference, neophobia, and high-fat cooking practices. The completion rate for the first telephone counseling call was 90%. Completion rates for the remaining three calls ranged from 79% to 86%. CONCLUSION: The recruitment and intervention methods of the Eat for Life study appear promising. The telephone intervention based on motivational interviewing is potentially useful for delivering dietary counseling. PMID- 11019765 TI - Partnering with African American churches to achieve better health: lessons learned during the Black Churches United for Better Health 5 a day project. AB - African Americans (AAs) are at increased risk for many diseases, including cancer, but health promotion efforts often fail to reach them. Effective partnerships can be established with African American churches to deliver health based interventions. In an NCI-funded study aimed at increasing fruit and vegetable consumption among rural AAs, investigators at three academic institutions and the North Carolina State Health Department partnered successfully with 50 churches to promote dietary change. This study adds to the increasing body of research in support of the African American church as an able partner in delivering health-based interventions. In conducting interventions and research through this channel, the health professional should gain support from regional secular associations; respect the power of the pastor; incorporate the strengths of the congregation; respect the mission of the church; establish open communication and develop trust; provide ample support and training to assure fidelity to interventions and integrity of data; and be patient and persevere. PMID- 11019766 TI - Community-based education in nutrition and cancer: the Por La Vida Cuidandome curriculum. AB - The Por La Vida (PLV) intervention model relies on community lay health advisors trained to conduct education sessions among members of their existing social networks. PLV Cuidandome was funded by the NCI to develop, implement, and evaluate the PLV model with respect to nutrition and cancer prevention, as well as early detection of breast and cervical cancers. The target population is the Latino community, for which substantial barriers to health care access exist. This article presents the curriculum that guides the sessions and describes its development, which was based on semi-structured interviews with Latina lay-health community workers to explore relevant attitudes and behaviors. Also key to the process was the work of the educational materials committee, whose members offered community representation as well as expertise in nutritional sciences, educational technologies, and community-based health promotion interventions and research. The 12-session curriculum's goal was to increase both the variety and the quality of fruits and/or vegetables consumed. It included information about consumption of fiber and fat in the importance of balance between energy intake and physical activity. The program has been well received. An ongoing study examines how it enhances nutrition and cancer prevention. PMID- 11019767 TI - Breast cancer education program based in Asian grocery stores. AB - Culturally and linguistically compatible university students were trained as community health educators to provide breast cancer education and screening information to shoppers at Asian grocery stores. Information about early detection of breast cancer was shared with 8,877 women, who reported speaking 40 different languages. Baseline surveys were completed by 1,202 women; 779 took part in the follow-up survey. The survey questions assessed baseline knowledge, attitudes, and screening behaviors regarding breast cancer, tested the efficacy of the intervention, and sought barriers to accessing screening services. Screening adherence at baseline was low, but reported screening compliance had increased by follow-up. This study confirms the cost-effectiveness of student health educators and Asian grocery store sites as venues to reach the diverse age, ethnic, and socioeconomic segments of the Asian community, while demonstrating the community's receptiveness to the dissemination of health information and introducing bilingual students to health education and research careers. PMID- 11019768 TI - The relationship of Internet use to depression and social isolation among adolescents. AB - The present study investigated whether higher levels of Internet use are associated with depression and social isolation among adolescents. Eighty-nine high school seniors were administered a questionnaire that measured low (less than 1 hour per day), moderate (1-2 hours per day), and high (more than 2 hours per day) Internet use; relationships with mother, father, and peers; and depression. Low Internet users, as compared with high users, reported better relationships with their mothers and friends. PMID- 11019769 TI - Scholastic achievement of adolescent refugees from Cambodia and Central America. AB - This study investigated the relationship of emotional disturbance and pre- and postmigration environment to the scholastic achievement of adolescent refugees of very different cultural backgrounds. One hundred fifty-two Central American and Cambodian students in six Canadian high schools, as well as their parents, were interviewed to assess the students' emotional problems (using the Youth Self Report and Child Behavior Checklist) and to determine the pre- and postmigration family environment. The findings indicated that the relationship between the emotional problems and scholastic achievement of teenaged refugees was tenuous. It was concluded that a connection between young refugees' symptomatology and their functional capacity should not be assumed. Nonetheless, certain pre- and postmigration variables, particularly trauma experienced in the homeland, seem to be associated with the academic achievement of some refugees. PMID- 11019770 TI - A comparative analysis of teenagers who smoke different cigarette brands. AB - This study analyzed and compared the survey responses of teenagers who smoke different cigarette brands. It was found that teen Marlboro and Camel smokers perceived themselves as having more stress in their lives as compared with teen Newport smokers. On average, Marlboro smokers were depressed or sad more often than were Camel or Newport smokers. Camel smokers were generally less interested in a peer stop-smoking program than were Newport or Marlboro smokers. Finally, these teen smokers, regardless of cigarette brand, had similar opinions about quitting. Given the differences across brands, more flexible approaches may be needed to address teenage smoking. PMID- 11019771 TI - Exposure to community violence in adolescence: trauma symptoms. AB - This research explored the relationship between exposure to violence and psychological stress among adolescents. It focused on the cumulative experience of recurring community violence during the high school years, differentiated victimization from witnessing violence, and examined four conceptually separate psychological symptoms of trauma (anger, anxiety, depression, and dissociation). Considerable exposure to recurring community violence, but also considerable variation in the amount of exposure, was found in the sample of 455 first-year students at an urban college. Exposure to recurring community violence was moderately correlated with the manifestation of psychological trauma symptoms. Further, being victimized and witnessing violence had independent (as well as overlapping) relationships with trauma symptoms. It was concluded that attempts to account for psychological stress among adolescents must include exposure to community violence. PMID- 11019772 TI - A longitudinal study of psychosocial changes among Thai adolescents participating in a Buddhist ordination program for novices. AB - Little is known about the effects of intense spiritual practice among teenage youth. In the present study, 170 adolescent males in Thailand were assessed regarding the immediate and long-term behavioral, emotional, and devotional consequences of participation in a six-week Buddhist monastic program. An additional 114 males of equivalent age, who were assessed in regard to changes following a demanding extracurricular English language course, served as controls. Their parents' perceptions were also examined. The results indicated that the changes reported by the youths in the ordination group were highly positive, enduring, and of a different pattern from that of the controls. The ratings of the two parent groups were also significantly different from each other, and analogous to those of their children. The practical and theoretical implications of the findings are discussed. PMID- 11019773 TI - Links between perceived parent characteristics and attachment variables for young women from intact families. AB - This study examined links between perceived parent characteristics and attachment variables for young women from intact families (biological parents still married to each other). One hundred fifty-four female college students served as subjects. They rated both parents using items derived from Secunda's (1992) descriptions of father characteristics, and also were assessed on measures of adult attachment, self-esteem, and interpersonal trust. Positive correlations were found between secure attachment and self-concept, good father characteristics, and doting mother characteristics. An insecure attachment pattern was associated with lower self-worth, less interpersonal trust, distant and demanding father characteristics, and absent mother characteristics. A fearful attachment pattern was associated with distant father and absent mother characteristics. A preoccupied attachment pattern was linked to absent, seductive, and demanding father characteristics and demanding mother characteristics. A dismissive attachment pattern was associated with distant father characteristics. Collectively, father characteristics related more strongly to an insecure attachment pattern, while mother characteristics related more strongly to a secure attachment pattern. PMID- 11019774 TI - Adolescent sexual behavior in two ethnic minority groups: a multisystem perspective. AB - Adolescents are at high risk for a number of negative health consequences associated with early and unsafe sexual activity, such as infection with HIV and other sexually transmitted diseases, as well as unintended pregnancy. In the present study, a multisystem model was applied to one adolescent sexual behavior, penile-vaginal intercourse. Nine hundred seven Black and Hispanic adolescents (aged 14 to 17 years) and their mothers were interviewed. Factors from three systems (self, family, and extrafamilial) that are influential in the lives of adolescents were evaluated using four outcome measures. Factors from most or all systems emerged as predictors of each outcome measure. A cumulative risk index suggested a linear relationship between the number of systems identified as being at risk and indicators of adolescent sexual behavior. The implications for prevention are discussed. PMID- 11019775 TI - Motivational needs of adolescent volunteers. AB - This study investigated the motivational needs of adolescent volunteers. The Volunteer Needs Profile (Francies, 1982) was administered to adolescents in a variety of volunteer settings, and data were examined using factor analysis. While there were similarities between the motivations of adolescent and older volunteers, some important differences were found. The findings are explored in the context of the adolescent developmental stage, and implications are discussed. PMID- 11019776 TI - Relationships between adolescents' attachment styles and family functioning. AB - This study examined the relationship between university students' perceptions of their familial attachment and the manner in which their families cope with life's difficulties. It was hypothesized that individuals with high levels of secure attachment would perceive their families as using more active coping strategies (e.g., mobilizing the family to deal with a problem and making efforts to acquire social support). The results supported this hypothesis. Further, individuals with high levels of anxious/ambivalent attachment perceived their families as using a passive appraisal coping strategy, possibly because of a desire to avoid confrontation for fear of disturbing family accord. The findings are discussed in terms of individuals' dispositional coping styles formed as a function of type of familial attachment, as well as situational factors that might serve to mitigate the use of their basic coping strategies. PMID- 11019777 TI - The blessing as a rite of passage in adolescence. AB - Central to the transition from adolescence to early adulthood is the transformation that takes place in the parent-child relationship, heretofore studied as emotional autonomy, psychological separation, and separation individuation. Blos (1985) suggested that individuation perhaps necessarily includes the confirmation of the child's adult status by the same-sex parent, called "the blessing." Of the 281 late adolescents in the present study, 71.5% indicated they had received some type of blessing from a parent and described the event as meaningful. Males were more likely to receive a blessing regarding instrumental traits, while females were more likely to receive a blessing regarding overall maturity, pubertal changes, or a specific rite of passage. These results offer support for Blos's position regarding the importance of this event to the young adult. PMID- 11019778 TI - Enhancing geometric reasoning. AB - Geometry is an important part of the mathematics curriculum. However, students are not demonstrating strong conceptual knowledge of this subject. The research of Van Hiele and Van Hiele-Geldof has focused on the concept of thinking levels in geometry and the role of instruction in raising levels of thinking. This paper describes a field trial of a supplemental geometry unit intended to raise Van Hiele thinking levels in a group of 23 eighth-grade students by having them become more adept at using higher order thinking skills. Sample questions assessing particular Van Hiele thinking levels and attitudes toward geometry, as well as field-tested activities yielding the most positive results, are presented. Educators can benefit from this application of the Van Hiele model of geometric thinking, since the thought processes involved in learning geometry are explained, along with teaching techniques and tools for assessment. By having teachers become more aware of their students' cognitive skills, attitudes, and misconceptions, teaching practices and student achievement can be enhanced. PMID- 11019779 TI - Targeting adolescent mothers with depressive symptoms for early intervention. AB - Infants of mothers with depressive symptoms show developmental delays if symptoms persist over the first 6 months of the infant's life, thus highlighting the importance of identifying those mothers for early intervention. In Study 1, mothers with depressive symptoms (n = 160) and mothers without depressive symptoms (n = 100) and their infants were monitored to identify variables from the first 3 months that predict which mothers would still be symptomatic at 6 months. A "dysregulation" profile was noted for the infants of depressed mothers, including lower Brazelton scores, more indeterminate sleep, and elevated norepinephrine, epinephrine, and dopamine levels at the neonatal period, and greater right frontal EEG activation, lower vagal tone, and negative interactions at the 3- and 6-month periods. A group of maternal variables from the neonatal and 3-month assessments accounted for 51% of the variance in the mothers' continuing depressive symptoms. These variables included greater right frontal EEG activation, lower vagal tone, and less positive interactions at 3 months, and elevated norepinephrine, serotonin, and cortisol levels at the neonatal stage. In Study 2, a similar sample of mothers with depressive symptoms (n = 160) and without depressive symptoms (n = 100) was recruited and followed to 3 months. Those symptomatic mothers who had values above (or below) the median (depending on the negative direction) on the predictor variables identified in Study 1 (taken from the first 3 months) were then randomly assigned to an intervention or a control group at 3 months. These groups were then compared with each other, as well as with the group without depressive symptoms, at 6 and 12 months. The intervention, conducted from 3 to 6 months, consisted of free day care for the infants and a rehab program (social, educational, and vocational) plus several mood induction interventions for the mothers, including relaxation therapy, music mood induction, massage therapy, and mother-infant interaction coaching. Although the mothers who received the intervention continued to have more depressive symptoms than did the nondepressed mothers, their interactions significantly improved and their biochemical values and vagal tone normalized. Their infants also showed more positive interations, better growth, fewer pediatric complications, and normalized biochemical values, and by 12 months their mental and motor scores were better than those of the infants in the control group. PMID- 11019780 TI - Transforming growth factor-alpha and epidermal growth factor receptor in colonic mucosa in active and inactive inflammatory bowel disease. AB - Transforming growth factor-alpha (TGF-alpha) is overexpressed in colonic carcinomas and promotes mucosal wound healing. It may be implicated in chronic inflammatory bowel disease (IBD). We analyzed the expression of TGF-alpha and its receptor, epidermal growth factor receptor (EGF-r), in the colonic mucosa of patients with Crohn's disease (CD) or ulcerative colitis (UC), in active or inactive stages, as compared with controls. Proteins and mRNA were detected in biopsies from the right and left colon and in surgical colonic specimens. Immunoblot analysis revealed TGF-alpha protein as a 29 kDa band. This band was normally expressed in uninvolved colonic mucosa of patients with CD or UC whether in active or inactive stages, but decreased or absent in involved mucosa of active IBD, even when TGF-alpha mRNA and EGF-r protein were detected. In the unaffected mucosa of CD, the intensity of TGF-alpha immunoreactivity was similar to that of controls in the right colon but stronger (P = 0.05) in the left colon. There was no TGF-alpha overexpression in dysplastic regions. In conclusion, in active IBD disease, the decreased TGF-alpha protein amount seems not only related to epithelial cell loss but reflects a down-regulation at least at the protein level. We speculate that TGF-alpha does not play a role within the active stage but may be implicated later in the repair process. PMID- 11019782 TI - Calcium-dependent and oncogenic IL-3 mRNA stabilization can be distinguished pharmacologically and by sequence requirements in the 3'UTR. AB - In interleukin-3 (IL-3)-dependent PB-3c mast cells, the normally short-lived IL-3 mRNA is stabilized upon calcium-ionophore treatment or following v-H-ras induced oncogenesis. We compared the underlying stabilization mechanisms by analysing the response to the post-transcriptionally acting drugs cyclosporin A (CsA), FK506 and SB202190. Stable IL-3 transcripts in the PB-3c-derived tumour cell line V2D1 decayed in response to CsA and FK506, but not in response to SB202190. Transcripts stabilized by elevating intracellular calcium levels in PB-3c cells were destabilized in response to all three drugs. In PB-3c cells, six AUUUA pentamers within the AU-rich element were sufficient to confer responsiveness to calcium-ionophore and CsA treatment. In V2D1 tumour cells, sensitivity to CsA required additional nucleotides flanking these pentamers. Our data suggest that IL-3 mRNA stabilization by either calcium-dependent or oncogenic pathways involves different intracellular mechanisms. PMID- 11019781 TI - Extracellular FGF-1 inhibits cytoskeletal organization and promotes fibroblast motility. AB - Previous efforts from this laboratory have established that acidic fibroblast growth factor (FGF-1), either added exogenously or secreted as a biologically active protein, induces a transformed phenotype in primary murine fibroblasts. Experimental studies described here demonstrate that constitutive exposure to extracellular FGF-I results in reduced cell attachment to multiple ligands, inhibition of cytoskeletal organization, and reduced collagen contraction, despite no detectable change in integrin cell surface expression. In addition, FGF-1-transduced fibroblasts demonstrated a > 10-fold increase in migration, an observation correlated with increased tyrosine phosphorylation of p125FAK and p130CAS. Collectively, these results suggest that FGF-1-induced fibroblast transformation includes the involvement of specific FGF receptor-mediated signal transduction cascades targeted to cytoskeletal and focal adhesion structures. PMID- 11019783 TI - Predisposition to colonic dysplasia is unaffected by continuous administration of insulin-like growth factor-I for twenty weeks in a rat model of chronic inflammatory bowel disease. AB - BACKGROUND: Insulin-like growth factor-I (IGF-I) is currently under evaluation for the treatment of a variety of chronic disease conditions. We investigated the safety of long-term IGF-I administration in a rat model of inflammatory bowel disease which predisposes to the development of dysplasia. METHODS: Chronic consumption of dextran sulphate sodium (DSS) by rats manifests a colitis with dysplastic features. Rats consumed 2% DSS for 4 weeks when pumps were implanted to deliver either vehicle or IGF-I for 15 or 20 weeks while rats continued to consume DSS. Features of colitis and dysplasia were assessed at kill. RESULTS: Compared to vehicle, 20 weeks IGF-I significantly increased body weight by 19% and total gut weight by 43%. Colonic crypt depth, proliferative compartment, labelling index, dysplasia, neoplasia and other indices of colitis were not significantly affected. CONCLUSIONS: Twenty weeks administration of IGF-I to rats induced growth of the intestine but did not affect the severity of experimentally induced colitis or the incidence or progression of colonic dysplasia. PMID- 11019784 TI - IL-2, its receptors, and bcl-2 and bax genes in normal, hyperplastic and carcinomatous human prostates: immunohistochemical comparative analysis. AB - The presence of interleukin-2 (IL-2) and its receptors (Ralpha, Rbeta, Rgamma), and their relationship with the products of bcl-2 and bax genes was studied in normal prostates, benign prostatic hyperplasia (BPH), and prostatic cancer (PC) by ELISA, Western blot, and immunohistochemistry. A comparative semiquantitative immunohistochemical study was also performed. For all the antibodies assayed, immunoreactions were found in the epithelium and some stromal cells in the three types of specimens studied. These immunoreactions were much more higher in PC samples than in normal prostates. In BPH, immunoreactions were similar to that of normal prostates (bax), similar to that of PC (IL-2 and its three receptors), or intermediate between that of normal prostates and that of PC (bcl-2). Immunoexpressions of IL-2 and its receptors were found in the epithelial basal cells and some stromal cell of normal prostates and might be related to the control of the proliferation-apoptosis equilibrium. The increased expressions of IL-2 and its receptors in the epithelium of prostates in BPH, associated with increased bcl-2 expression which would account for the decrease in the apoptosis index that has been reported in this disorder. The increased expression of both bcl-2 and bax in PC might be involved in the higher apoptosis index reported in PC specimens. Since IL-2 administration seems to have an anti-tumour effect, the increased expression of this interleukin in BPH and PC could be interpreted as an attempt to hinder cell proliferation which would only be efficient at high doses. PMID- 11019785 TI - A conjugate composed of nerve growth factor coupled to a non-toxic derivative of Clostridium botulinum neurotoxin type A can inhibit neurotransmitter release in vitro. AB - Nerve growth factor (NGF) receptor binding, internalisation and transportation of NGF has been identified as a potential route of delivery for other molecules. A derivative of Clostridium botulinum neurotoxin type A (LHN) that retains catalytic activity but has significantly reduced cell-binding capability has been prepared and chemically coupled to NGF. Intact clostridial neurotoxins potently inhibit neurotransmitter release at the neuromuscular junction by proteolysis of specific components of the vesicle docking/fusion complex. Here we report that the NGF-LHN/A conjugate, when applied to PC12 cells, significantly inhibited neurotransmitter release and cleaved the type A toxin substrate. This work represents the successful use of NGF as a targeting moiety for the delivery of a neurotoxin fragment. PMID- 11019786 TI - Progress in desmin-related myopathies. AB - Desmin-related myopathies are sporadic and familial neuromuscular conditions of considerable clinical heterogeneity uniformly marked by the pathologic accretion of desmin, often in a filamentous fashion. A large variety of other proteins, some of them cytoskeletal, also accrue. Morphologically, two types may be distinguished, one characterized by inclusions such as cytoplasmic and spheroid bodies or desmin-dystrophin plaques and another marked by granulofilamentous material. The genetic spectrum of desmin-related myopathies is quite diverse in that missense mutations and deletions in the desmin gene and a missense mutation in the alpha-B crystallin gene have been detected and several genes on other chromosomes have been mapped; the encoded protein products of these genes, however, are unknown. Accumulation of desmin and other proteins appears to be due to impaired nonlysosomal proteolysis. Mutant desmin that appears to be hyperphosphorylated seems to act as a seed protein for filament aggregation, inducing formation of inclusions and granulofilamentous material in these conditions. This condition is part of the group of disorders known as "surplus protein myopathies." PMID- 11019787 TI - Methotrexate treatment protocols and the central nervous system: significant cure with significant neurotoxicity. AB - Methotrexate can influence the central nervous system through several metabolic toxic pathways. These effects can be categorized as immediate, acute to subacute, or chronic neurologic syndromes. The acute to subacute syndrome occurs frequently in acute lymphoblastic leukemia treatment protocols, generally manifesting with focal neurologic signs and changes seen on magnetic resonance imaging and single photon emission computed tomography. While in some patients the neurotoxicity is transient and benign and allows for continuation of chemotherapy, in others it can be quite severe and debilitating, leading to permanent neurologic deficits. The need to modify the treatment protocols when neurotoxicity appears is not fully established. It is also unknown whether the use of sufficient amounts of leucovorin can overcome the toxic effects of the drug. PMID- 11019789 TI - Possible mechanisms in infants for selective basal ganglia damage from asphyxia, kernicterus, or mitochondrial encephalopathies. AB - Magnetic resonance imaging and neuropathologic studies have demonstrated remarkably selective patterns of injury to subregions of the basal ganglia in children. Examples are kernicterus and certain mitochondrial encephalopathies, which cause selective injury to the globus pallidus, and near-total perinatal asphyxia, which causes lesions in the putamen and thalamus. To explain the differential vulnerability of nuclei within millimeters of each other, we hypothesize that their locations within the neurotransmitter-specific circuitry of the basal ganglia motor loop are important. In severe hypoxic-ischemic encephalopathy, excitatory glutamatergic pathways into the putamen and thalamus are overactive, but the globus pallidus might be protected because its activity is silenced by inhibitory neuronal activity. In contrast, the relatively high resting neuronal activity in the globus pallidus might make it more vulnerable to less intense, subacute oxidative stresses from mitochondrial toxins such as bilirubin or from genetic mitochondrial disorders. This hypothesis has implications for designing neuroprotective therapies and for treating associated chronic movement disorders. PMID- 11019788 TI - Cognitive outcome of children with early-onset hemiparesis. AB - The aim of this study was to examine the cognitive outcome of children with congenital and acquired early-onset unilateral brain lesions associated with hemiparesis. The neuropsychologic evaluation was done using the NEPSY test battery on 37 children with hemiparesis and 13 sex- and age-matched healthy controls. Compared to the controls, children with left-sided brain lesions had significant delay in phonologic and language functions, while children with right sided brain lesions performed more poorly in visual and spatial skills and in somatosensory functions. There were more left-handed children in the former (6 of 23) than in the latter (1 of 14) group. There was no significant difference in cognitive outcome between children with congenital and acquired lesions. The cognitive outcome of boys and that of children with active epilepsy was more affected. Overall, the patients showed impairment in many cortical functions and diffuse cognitive delay compared to controls and the side of lesion, active epilepsy, and male gender were significant factors in predicting cognitive outcome. PMID- 11019790 TI - Diffusion-weighted imaging in neonatal cerebral infarction: clinical utility and follow-up. AB - We describe the clinical utility of echo-planar diffusion-weighted imaging in neonatal cerebral infarction. Eight full-term neonates aged 1 to 8 days referred for neonatal seizures were studied. Patients were followed for a mean of 17 months with detailed neurologic examinations at regular intervals. Head computed tomography (CT) and conventional magnetic resonance (MRI) and diffusion-weighted images were obtained. Percent lesion contrast was evaluated for 19 lesions on T2 weighted and diffusion-weighted images. Follow-up conventional MRIs were obtained in seven patients. The findings on diffusion-weighted imaging were correlated with CT and conventional MRI findings as well as with short-term neurodevelopmental outcome. Four patients had focal cerebral infarctions. Four patients had diffuse injury consistent with hypoxic-ischemic encephalopathy. Percent lesion contrast of all 19 lesions was significantly higher on diffusion weighted images when compared with T2-weighted images. In five patients, there were lesions visualized only with diffusion-weighted imaging. In all patients, there was increased lesion conspicuity and better definition of lesion extent on the diffusion-weighted images compared with the CT and T2-weighted MR images. In seven of eight patients follow-up imaging confirmed prior infarctions. Short-term neurologic outcome correlated with the extent of injury seen on the initial diffusion-weighted imaging scans for all patients. Diffusion-weighted imaging is useful in the evaluation of acute ischemic brain injury and seizure etiology in neonates. In the acute setting, diffusion-weighted imaging provides information not available on CT and conventional MRI. This information correlates with short term clinical outcome. PMID- 11019791 TI - Crying abnormalities in congenital hypothyroidism: preliminary spectrographic study. AB - The aim of this preliminary study was to evaluate the acoustic patterns of the cries of hypothyroid newborns at the time of diagnosis and after the beginning of therapy. Cries were recorded at the nursery of the San Raffaele Hospital, Milan, Italy from 12 full-term subjects (three boys and nine girls) affected by congenital hypothyroidism. Results show that untreated hypothyroid infants at first recording had fewer voiceless and partially voiced cries than normal controls. The percent distribution of this pattern did not change at the second recording after the onset of substitutive therapy. Also, untreated hypothyroid infants had many more cry units showing a vibrato contour than did controls, and this pattern did not change after the onset of treatment. Starting, maximum, minimum, and end frequencies measured on the fundamental were significantly lower in the hypothyroid sample. Four hypothyroid subjects recorded before therapy and within 4 weeks after therapy onset significantly augmented their fundamental frequency parameters; however, in 25% of the sample, sound parameters remained unaltered after 3 or more weeks of treatment. To our knowledge the present preliminary study is the first one performed on follow-up of hypothyroid newborns and indicates that both central and peripheral damage might influence the pattern of crying in untreated hypothyroid infants. PMID- 11019792 TI - Managing allegations of scientific misconduct: a guidance document for editors. PMID- 11019793 TI - Neurometabolic testing in developmental delay. AB - The goal of this study was to examine the usefulness of neurometabolic testing in patients with unexplained developmental delay. We included 118 patients from ages 3 months to 13 years. The evaluation was conducted according to a protocol that called for stepwise investigations with the intention to minimize unnecessary tests. Clinical and neuroradiologic abnormalities were used as guidelines for the type and extent of the neurometabolic work-up. Based on our results, a diagnosis for a neurometabolic disorder was established in 16 patients (13.6%). Findings regarding abnormal metabolites (amino acids, ammonia, lactate, pyruvate, urine organic acids) did not lead to a diagnosis but to further investigations. Abnormalities in lysosomal enzymes, very-long-chain fatty acids, and urine mucopolysaccharides were definitive in establishing a diagnosis. In conclusion, in children with slowly progressing development with no apparent cause, there is a need for guidelines regarding the type of patients who need to be tested and the kind and extent of neurometabolic diagnostic work-up. PMID- 11019794 TI - Unilateral periventricular nodular heterotopia associated with diffuse areas of cerebral functional abnormalities. AB - A 17-year-old boy with polymorphic simple and complex partial seizures is described. Magnetic resonance imaging revealed a unilateral periventricular nodular heterotopia near the occipital ventricular right horn. Interictal and ictal electroencephalographic recordings showed bilateral specific epileptiform anomalies in the occipital region and asynchronous slow waves in frontal areas. Single photon emission computed tomography documented a reduction in regional cerebral blood flow in an area of the left occipital cortex and a symmetric increase in tracer uptake in the frontal lobes. The neuropsychologic assessment revealed a dysfunction of the frontal associative areas. Data collected led the authors to suspect a more diffuse cortical dysfunction than the nodular heterotopia revealed on magnetic resonance imaging. PMID- 11019795 TI - Long-term follow-up of auditory agnosia as a sequel of herpes encephalitis in a child. AB - We report a pediatric patient with auditory agnosia as a sequel of herpes encephalitis. His early development was completely normal. He uttered three words at 12 months old. Disease onset was 1 year and 2 months of age. He was discharged from the hospital seemingly with no sequel; however, he could not recover his intelligible words even at age 2 years. He was diagnosed as having auditory agnosia caused by bilateral temporal lobe injury. We began to train him at once, individually and intensively. Adult patients with pure auditory agnosia followed by two episodes of temporal lobe infarction have impairment in central hearing but not inner language. Therefore, they can communicate by reading and writing. Moreover, impairment in hearing is not always severe and is often transient. However, despite long-term (more than 15 years) energetic education and almost normal intellectual ability (Performance IQ of Wechsler Intelligence Scale for Children-Revised was 91), our patient's language ability was extremely poor. Cerebral plasticity could not work fully on our patient, whose bilateral temporal lobe was severely injured in early childhood. The establishment of a systematic training method in such patients is an urgent objective in this field. PMID- 11019796 TI - Electrodiagnosis of infantile botulism. PMID- 11019797 TI - Benign macrocephaly: a common cause of big heads in the first year. PMID- 11019798 TI - Tics in Tourette syndrome and botulinum toxin. PMID- 11019799 TI - Syncope and seizures: it is time for evidence! PMID- 11019800 TI - Web alert. Cell signalling and gene regulation. PMID- 11019801 TI - Ethylene signaling: from mutants to molecules. AB - The past decade has been incredibly productive for ethylene researchers. Major components in the ethylene signaling pathway in plants have been identified and characterized. The past year's contributions include the crystallographic analysis of the Arabidopsis ETR1 receiver domain, antisense studies of the tomato ethylene receptor genes LeETR4 and NR, and the cloning and functional characterization of several Arabidopsis EREBP-related transcription activators and repressors, and of an EIN3-ortholog of tobacco. Additional evidence for the interconnection of the ethylene and auxin responses was provided by the cloning and characterization of Arabidopsis NPH4. Finally, the first discovery of ethylene responsiveness in an animal species implied a more universal role for ethylene than previously thought. PMID- 11019802 TI - Receptor kinase activation and signal transduction in plants: an emerging picture. AB - Plant receptor kinases play key roles in the cell-cell recognition process during development, defense against pathogens, and self incompatibility. Recent identification of potential ligand molecules and downstream signaling components, together with biochemical studies on receptor-complex formation, have revealed an emerging picture of receptor-kinase activation and signal transduction in plants. PMID- 11019803 TI - Cell-cell signaling in the self-incompatibility response. AB - Significant progress towards understanding the molecular basis of self recognition in the self-incompatibility response of Brassica has been made during the past two years. The highly polymorphic molecules that determine the specificity of this interaction in the pollen and stigma have been identified. The structural features of these molecules suggest that a ligand-receptor-kinase interaction triggers the initiation of a signaling cascade within the stigma epidermis and the subsequent arrest of self-pollination. PMID- 11019804 TI - Gibberellin signal transduction. AB - Recent studies using biochemical and genetic approaches have identified a number of components, including several negative regulators, of the gibberellin (GA) signal transduction pathway in higher plants. The basal state of GA signaling is likely to be repressive, and the GA signal seems to activate the pathway by de repression to allow GA-stimulated growth and development. PMID- 11019805 TI - Protein degradation in signaling. AB - Recent studies have linked proteolysis by the ubiquitin/proteasome pathway to a variety of signaling pathways in higher plants. These links were uncovered by characterization of mutants altered in signaling or by targeted disruption of components of the proteolytic pathway. Significant advances have recently revealed connections between proteolysis and hormone responses, light perception, environmental adaptation, and floral development. PMID- 11019806 TI - The COP9 signalosome: from light signaling to general developmental regulation and back. AB - The COP9 signalosome has eight core subunits that are highly conserved between plants and animals. Some of the subunits in Arabidopsis are found in forms that are independent of the complex. The COP9 complex is essential for animal development. The COP9 signalosome may have both an evolutionary and a physical relationship with both the regulatory lid of the proteasome and eIF3. PMID- 11019807 TI - Opening up the communication channels: recent insights into plasmodesmal function. AB - The past year has seen significant advances in our understanding of the function and regulation of plasmodesmata. Notably, we have learned that plasmodesmata undergo dynamic changes during development and may participate in long-range communication through the transmission of RNA signals. Biochemical studies have enriched our understanding of a putative plasmodesmal receptor and of plant factors involved in viral cell-to-cell movement. PMID- 11019808 TI - Regulatory 14-3-3 protein-protein interactions in plant cells. AB - Many biological roles for plant 14-3-3 proteins have been suggested in recent months. The most significant of these include roles in the import of nuclear encoded chloroplast proteins, in the assembly of transcription factor complexes and in the regulation of enzyme activity in response to intracellular signal transduction cascades. PMID- 11019809 TI - Vascular development: tracing signals along veins. AB - The plant hormone auxin has been implicated in vascular development, but the molecular details of patterned vascular differentiation have remained elusive. Research in the past year has identified new genes that control vascular patterning, and auxin transport and perception. New experimental strategies have been employed to study vascular development. Together, these findings have generated a conceptual framework and experimental tools for the exploration of vascular-tissue patterning at the molecular level. PMID- 11019810 TI - Non-linear signaling for pattern formation? AB - Developmental pathways are often simplified as linear cascades of gene activation caused by initial signaling events. Recent data, however, support a more complex view in which non-linear responses to signals are important for pattern formation. First, a positive feedback loop may regulate cellular patterning in the Arabidopsis epidermis. Second, an asymmetric auxin distribution that organizes pattern in the distal root tip may be established by positive feedback. PMID- 11019811 TI - The control of flowering by vernalization. AB - The process by which vernalization, the exposure of a germinating seed or a juvenile plant to a prolonged period of low temperature, promotes flowering in the adult plant has remained a mystery for many years. The recent isolation of one of the key genes involved in vernalization, FLOWERING LOCUS C, has now provided an insight into the molecular mechanism involved, including the role of DNA methylation. PMID- 11019812 TI - A genomic perspective on plant transcription factors. AB - Data from the Arabidopsis genome project suggest that more than 5% of the genes of this plant encode transcription factors. The necessity for the use of genomic analytical approaches becomes clear when it is considered that less than 10% of these factors have been genetically characterized. A variety of tools for functional genomic analyses in plants have been developed over the past few years. The availability of the full complement of Arabidopsis transcription factors, together with the results of recent studies that illustrate some of the challenges to their functional characterization, now provides the basic framework for future analyses of transcriptional regulation in plants. PMID- 11019813 TI - Starvation increases the amount of pyruvate dehydrogenase kinase in several mammalian tissues. AB - Covalent modification of the pyruvate dehydrogenase complex provides an important regulatory mechanism for controlling the disposal of glucose and other compounds metabolized to pyruvate. Regulation of the complex by this mechanism is achieved in part by tissue-specific expression of the genes encoding isoenzymes of pyruvate dehydrogenase kinase (PDK). Starvation is known from our previous work to increase PDK activity of heart and skeletal muscle by increasing the amount of PDK isoenzyme 4 (PDK4) present in these tissues. This study demonstrates that increased expression of both PDK4 and PDK2 occurs in rat liver, kidney, and lactating mammary gland in response to starvation. PDK4 and PDK2 message levels were also increased by starvation in the two tissues examined (liver and kidney), suggesting enhancement of gene transcription. Changes in PDK2 message and protein were of similar magnitude, but changes in PDK4 message were greater than those in PDK4 protein, suggesting regulation at the level of translation. In contrast to these tissues, starvation had little or no effect on PDK2 and PDK4 protein in brain, white adipose tissue, and brown adipose tissue. Nevertheless, PDK4 message levels were significantly increased in brain and white adipose tissue by starvation. The findings of this study indicate that increased expression of PDK isoenzymes is an important mechanism for bringing about inactivation of the pyruvate dehydrogenase complex during starvation in many but not all tissues of the body. The absence of this mechanism preserves the capacity of neuronal tissue to utilize glucose for energy during starvation. PMID- 11019814 TI - A new method for the analysis of urinary vitamin E metabolites and the tentative identification of a novel group of compounds. AB - There is currently interest in measuring urinary metabolites of vitamin E. It has been suggested that alpha-to-copheronolactone (alphaTL), with an oxidized chroman ring, may be an indicator of in vivo oxidative stress and 2,5,7,8-tetramethyl 2(2'-carboxyethyl)-6-hydroxychroman (alpha-CEHC), with a shortened side chain but intact hydroxychroman ring, may provide a measure of adequate or excess vitamin E status. To date, methods in the literature have tended to concentrate on the estimation of single metabolites. We describe the establishment and validation of a relatively simple and reproducible method to extract and quantitate a range of vitamin E metabolites using 0.5 ml of human urine. The vitamin E metabolites were extracted from urine using solid phase extraction cartridges, deconjugated enzymatically, and analyzed using gas chromatography-mass spectrometry. Using this method we have identified alphaTL and the CEHC metabolites derived from alpha-, delta-, and gamma-tocopherol. In addition we have tentatively identified a novel group of vitamin E metabolites, which are related to the CEHCs but have three extra carbons in the side chain. The possibility of the artifactual oxidation of alpha-CEHC to alphaTL during the assay procedure is also discussed. PMID- 11019815 TI - MnII is not a productive substrate for wild-type or recombinant lignin peroxidase isozyme H2. AB - The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene promoter was used to drive the homologous expression of the lignin peroxidase (LiP) isozyme H2 gene in primary metabolic cultures of Phanerochaete chrysosporium. The molecular mass, pI, and optical absorption spectra of purified recombinant LiPH2 (rLiPH2) were essentially identical to those of wild-type LiPH2 (wtLiPH2). wtLiPH2 was prepared by growing cells in the absence of MnII, conditions under which P. chrysosporium manganese peroxidase (MnP) is not expressed, ensuring that wtLiPH2 was not contaminated with MnP. The kinetics of veratryl alcohol (VA) oxidation were essentially identical for rLiPH2 and wtLiPH2. The rLiPH2, wtLiPH2, and wild-type LiP isozyme H8 (wt-LiPH8) enzymes were used to reexamine previous claims that LiPH2 can oxidize Mn" at a rate sufficient to promote catalytic turnover of the enzyme. Our results demonstrate that rLiPH2, wtLiPH2, and LiPH8 do not turn over under steady-state conditions, when MnII is the sole reducing substrate. Furthermore, transient-state kinetic analyses show that the reduction rate of the catalytic intermediate, LiP compound I, by VA was at least 2 x 10(3)-fold higher than the rate of reduction in the presence of MnII. No reduction of LiP compound II was observed in the presence of MnII. In contrast to previous claims, these data strongly suggest that MnII is not a productive substrate for LiPH2 or LiPH8. PMID- 11019816 TI - Decavanadate inhibits catalysis by ribonuclease A. AB - Pentavalent organo-vanadates have been used extensively to mimic the transition state of phosphoryl group transfer reactions. Here, decavanadate (V(10)O(28)6-) is shown to be an inhibitor of catalysis by bovine pancreatic ribonuclease A (RNase A). Isothermal titration calorimetry shows that the Kd for the RNase A decavanadate complex is 1.4 microM. This value is consistent with kinetic measurements of the inhibition of enzymatic catalysis. The interaction between RNase A and decavanadate has a coulombic component, as the affinity for decavanadate is diminished by NaCl and binding is weaker to variant enzymes in which one (K41A RNase A) or three (K7A/R10A/K66A RNase A) of the cationic residues near the active site have been replaced with alanine. Decavanadate is thus the first oxometalate to be identified as an inhibitor of catalysis by a ribonuclease. Surprisingly, decavanadate binds to RNase A with an affinity similar to that of the pentavalent organo-vanadate, uridine 2',3'-cyclic vanadate. PMID- 11019817 TI - Mouse group X secretory phospholipase A2 induces a potent release of arachidonic acid from spleen cells and acts as a ligand for the phospholipase A2 receptor. AB - Group X secretory phospholipase A2 (sPLA2-X) has recently been shown to possess a powerful potency for releasing arachidonic acid from cell membrane phospholipids. Here, we report the purification of mouse pro- and mature forms of sPLA2-X, as well as its expression and biological functions. Purified pro-sPLA2-X was found to possess a propeptide of 11 amino acid residues attached at the NH2-terminals of the mature protein, and showed as little as 8% of the PLA2 activity of the mature form. Limited proteolysis of pro-sPLA2-X with trypsin resulted in the appearance of the mature form with a concomitant increase in PLA2 activity, suggesting a requirement of proteolytic removal of the propeptide for the optimal activity. The expression of sPLA2-X mRNA was detected in various tissues including the lung, thymus, and spleen, and immunohistochemical analysis revealed its expression in splenic macrophages. In the spleen cells, mature sPLA2-X elicited a prompt release of arachidonic acid with significant production of prostaglandin E2 more efficiently than group IB and IIA sPLA2s. In addition, sPLA2-X was identified as a high-affinity ligand for both native and recombinant form of mouse PLA2 receptor (PLA2R). However, there was no significant difference in the sPLA2-X-induced arachidonic acid release responses in the spleen cells between wild-type and PLA2R-deficient mice. These findings strongly suggest that sPLA2-X possesses two distinct biological functions in mice: it elicits a marked release of arachidonic acid from membrane phospholipids leading to the production of lipid mediators based on its enzymatic potency, and it acts as a natural ligand for the PLA2R that has been shown to play a critical role in the production of inflammatory cytokines during endotoxic shock. PMID- 11019818 TI - Photoaffinity labeling and site-directed mutagenesis of rat squalene epoxidase. AB - Squalene epoxidase (SE) (EC 1.14.99.7) is a flavin-requiring, non-cytochrome P 450 oxidase that catalyzes the conversion of squalene to (3S)-2,3-oxidosqualene. Photolabeling and site-directed mutagenesis were performed on recombinant rat SE (rrSE) to elucidate the location and roles of active-site residues important for catalysis. Two new benzophenone-containing analogs of NB-598, a nanomolar inhibitor of vertebrate SE, were synthesized in tritium-labeled form. These photoaffinity analogs (PDA-I and PDA-II) became covalently attached to SE when irradiated at 360 nm. Lys-C digestion and HPLC purification of [3H]PDA-I-labeled rrSE resulted in isolation of a single major peptide. MALDI-TOF mass spectrometry of this peptide indicated a covalent adduct between PDA-I and a tripeptide, Asp Ile-Lys, beginning at Asp-426 of rat SE. Based on the labeling results, three mutant constructs were made. First, the D426A and K428A constructs showed a 5- to 8-fold reduction in SE activity compared with wild-type enzyme, while little change was observed in the I427A mutant. Second, a set of five mutant constructs was prepared for the conserved region based on the structure of the flavoprotein p-hydroxybenzoate hydroxylase (PHBH). Compared with wild-type, D284A and D407A showed less than 25% SE activity. This reduction also appeared to correlate with reduced affinity of the mutant proteins for FAD. Finally, each of the seven Cys residues of rrSE were individually mutated to Ala. Three Cys substitutions had no effect on SE activity, and substitutions at Cys-500 and Cys-533 showed a 50% lower SE activity. Mutations at Cys-490 and Cys-557 produced proteins with negligible SE activity, implicating these residues as being either structurally or catalytically essential. Chemical modification of wildtype and Cys mutants with a thiol-modifying reagent support the existence of a disulfide bond between Cys-490 and Cys-557. PMID- 11019819 TI - Primary structure characterization of a Rhodocyclus tenuis diheme cytochrome c reveals the existence of two different classes of low-potential diheme cytochromes c in purple phototropic bacteria. AB - The complete amino acid sequence of a 26-kDa low redox potential cytochrome c-551 from Rhodocyclus tenuis was determined by a combination of Edman degradation and mass spectrometry. There are 240 residues including two heme binding sites at positions 41, 44, 128, and 132. There is no evidence for gene doubling. The only known homolog of Rc. tenuis cytochrome c-551 is the diheme cytochrome c-552 from Pseudomonas stutzeri which contains 268 residues and heme binding sites at nearly identical positions. There is 44% overall identity between the Rc. tenuis and Ps. stutzeri cytochromes with 10 internal insertions and deletions. The Ps. stutzeri cytochrome is part of a denitrification gene cluster, whereas Rc. tenuis is incapable of denitrification, suggesting different functional roles for the cytochromes. Histidines at positions 45 and 133 are the fifth heme ligands and conserved histidines at positions 29, 209, and 218 and conserved methionines at positions 114 and 139 are potential sixth heme ligands. There is no obvious homology to the low-potential diheme cytochromes characterized from other purple bacterial species such as Rhodobacter sphaeroides. There are therefore at least two classes of low-potential diheme cytochromes c found in phototrophic bacteria. There is no more than 11% helical secondary structure in Rc. tenuis cytochrome c 551 suggesting that there is no relationship to class I or class II c-type cytochromes. PMID- 11019820 TI - Metabolism of alpha-D-[1,2-13C]glucose pentaacetate and alpha-D-glucose penta[2 13C]acetate in rat hepatocytes. AB - Hepatocytes from fed rats were incubated for 120 min in the presence of alpha-D [1,2-13C]glucose pentaacetate (1.7 mM), both D-[1,2-13C]glucose (1.7 mM) and acetate (8.5 mM), alpha-D-glucose penta[2-13C]acetate (1.7 mM), or D-[1,2 13C]glucose (8.3 mM). The amounts of 13C-enriched L-lactate and D-glucose and those of acetate and beta-hydroxybutyrate recovered in the incubation medium were comparable under the first two experimental conditions. The vast majority of D glucose isotopomers consisted of alpha- and beta-D[1,2-13C]glucose. The less abundant single-labeled isotopomers of D-glucose were equally labeled on each C atom. The output of 13C-labeled L-lactate, mainly L-[2-13C]lactate and L-[3 13C]lactate, was 1 order of magnitude lower than that found in hepatocytes exposed to 8.3 mM D-[1,2-13C]glucose, in which case the total production of the single-labeled species of D-glucose was also increased and that of the C3- or C4 labeled hexose was lower than that of the other 13C-labeled isotopomers. In cells exposed to alpha-D-glucose penta[2-13C]acetate, the large majority of 13C atoms was recovered as [2-13C]acetate and, to a much lesser extent, beta hydroxybutyrate labeled in position 2 and/or 4. Nevertheless, L-[2-13C]lactate, L [3-13C]lactate, and single-labeled D-glucose isotopomers were also produced in amounts higher or comparable to those found in cells exposed to alpha-D-[1,2 13C]glucose pentaacetate. However, a modest preferential labelling of the C6-C5 C4 moiety of D-glucose, relative to its C1-C2-C3 moiety, and a lesser isotopic enrichment of the C3 (or C4), relative to that of C1 (or C6) and C2 (or C5), were now observed. These findings indicate that, despite extensive hydrolysis of alpha D-glucose pentaacetate (1.7 mM) in the hepatocytes, the catabolism of its D glucose moiety is not more efficient than that of unesterified D-glucose, tested at the same molar concentration (1.7 mM) in the presence of the same molar concentration of unesterified acetate (8.5 mM), and much lower than that found at a physiological concentration of the hexose (8.3 mM). The present results also argue against any significant back-and-forth interconversion of D-glucose 6 phosphate and triose phosphates, under conditions in which sizeable amounts of D glucose are formed de novo from 13C-enriched Krebs cycle intermediates generated from either D-[1,2-13C]glucose or [2-13C]acetate. PMID- 11019821 TI - Rapid effects of stretched myocardial and vascular cells on gene expression of neonatal rat cardiomyocytes with emphasis on autocrine and paracrine mechanisms. AB - Passive stretch of the heart has a direct effect on cardiomyocytes and other cell types including cardiac fibroblasts, endothelial cells, and vascular smooth muscle cells (VSMCs). Cardiomyocytes are targets for the action of peptide growth factors found in myocardium, suggesting an autocrine or paracrine model of the hypertrophic process. In this study we examined stretch-dependent cellular communication between cardiomyocytes, cardiac fibroblasts, endothelial cells, and VSMCs. Stationary cardiomyocytes were incubated with stretch-conditioned medium (CM0-CM60) derived from stretched (for 0-60 min) cardiomyocytes, cardiac fibroblasts, endothelial cells, and VSMCs. The expression levels of protooncogenes (as c-fos, c-jun, and fra-1) were measured, and as an indication of a hypertrophic response the expression of atrial natriuretic peptide (ANP) was measured. Stationary cardiomyocytes that have been incubated for 30 min with CM from stretched (for 0-60 min) cardiomyocytes, cardiac fibroblasts, endothelial cells, and VSMCs showed distinct gene expression patterns that were time dependent and cell-type specific. In stationary cardiomyocytes, CM derived from stretched cardiomyocytes caused decreased c-fos and fra-1 expression by 37 and 20%, respectively (CM30), elevated c-jun expression by 20% (CM45-CM60), and increased ANP expression by 106% (CM45). CM derived from stretched cardiac fibroblasts caused increased c-fos expression by 41% (CM60), no significant changes in c-jun expression, and increased fra-1 and ANP expression by 39 and 20%, respectively (CM45). CM derived from stretched VSMCs induced an initial decrease in c-fos expression followed by an increase of 13% (CM45) and induced increased c-jun, fra-1, and ANP expression by 39, 24, and 22%, respectively. CM15 CM60 derived from stretched endothelial cells caused decreased c-fos, c-jun and fra-1 expression by 20, 25, and 25%, respectively, and increased ANP expression by 18%. Our data indicate that gene expression of cardiomyocytes in stretched myocardium is regulated by mediators released by cardiomyocytes, cardiac fibroblasts, endothelial cells, and VSMCs. This observation emphasizes the involvement of nonmyocyte cells in the early stages of cardiomyocyte hypertrophy caused by cardiac stretch. PMID- 11019822 TI - Mechanism of activation of cAMP-dependent protein kinase: in Mucor rouxii the apparent specific activity of the cAMP-activated holoenzyme is different than that of its free catalytic subunit. AB - Kinetic constants for peptide phosphorylation by the catalytic subunit of the dimorphic fungus Mucor rouxii protein kinase A were determined using 13 peptides derived from the peptide containing the basic consensus sequence RRASVA, plus kemptide, S6 peptide, and protamine. As a whole, although with a greater Km, the order of preference of the peptides by the M. rouxii catalytic subunit was similar to the one displayed by mammalian protein kinase A. Particularly significant is the replacement of serine by threonine in the basic peptide RRATVA, which impaired its role as a substrate of M. rouxii catalytic subunit. Mucor rouxii protein kinase A is a good model in which to study the mechanism of activation since cAMP alone is not enough to promote activation and dissociation. Four peptides were selected for the study of holoenzyme activation under conditions in which the enzymatic activity was not proportional to the holoenzyme concentration: RRASVA, RRRRASVA, KRRRLSSRA (S6 peptide), and LRRASLG (kemptide); protamine was used as reference. Differential activation degree was observed depending on the peptide used and on cAMP concentration. Ratios of activity between different substrates displayed by the holoenzyme under the above conditions did not reflect the one expected for the free catalytic subunit. The degree of inhibition of the holoenzyme activity by an active peptide derived from the thermostable protein kinase inhibitor was dependent on the substrate used and on the holoenzyme concentration, while it was found to be independent of these two parameters for free catalytic subunit. Polycation modulation of holoenzyme activation by cAMP was also dependent on the polycation itself and on the peptide used as substrate. The observed kinetic differences between holoenzyme and free catalytic subunit were decreased or almost abolished when working at low enzyme or at high cAMP concentrations. Two hypotheses compatible with the results are discussed: substrate participation in the dissociation process and/or holoenzyme activation without dissociation. PMID- 11019823 TI - Acid dissociation constants of bilirubin and related carboxylic acid compounds in bile salt solutions. AB - Bilirubin, the yellow-orange tetrapyrrole pigment of jaundice, is essentially insoluble in pure water, but is much more soluble in solutions of bile salts such as sodium taurocholate. The biophysical chemistry of bilirubin in bile salt solutions is affected by changes in the pH of the solution in the range 5-9, suggesting that interactions with bile salt molecules and micelles may alter the acidity of the pigment. We have examined this possibility by determining the apparent pKa values for a series of carboxyl 13C-enriched model compounds, including the bilirubin analog mesobilirubin XIIIalpha, in solutions of sodium taurocholate and sodium taurodeoxycholate. Apparent pKa values were determined by 13C NMR titrations in dimethyl sulfoxide-water mixtures. The results show that the acidity of all compounds is decreased, or pKa increased, in micellar bile salt solution relative to pure water and that the effect is greatest for the larger, less water-soluble compounds. We have proposed a model to explain these results and discussed the implications of these findings for the biophysical chemistry of bilirubin in bile. PMID- 11019824 TI - A modified method of islet isolation preserves the ability of pancreatic islets to increase protein O-glycosylation in response to glucose and streptozotocin. AB - An important link has recently been shown in vivo between beta-cell O-linked protein glycosylation and beta-cell apoptosis, with hyperglycemia having been demonstrated to reversibly increase beta-cell O-linked protein glycosylation by providing substrate for the glucosamine pathway. In contrast, the same study showed that the administration of streptozotocin to rats prior to the induction of hyperglycemia results in irreversible increases in O-glycosylation and subsequent beta-cell apoptosis. In light of these data, we investigated beta-cell O-glycosylation in vitro by exposing isolated rat islets to high glucose, glucosamine, or streptozotocin and analyzing the pattern of O-glycosylated proteins present. All three compounds acutely increased O-glycosylation of a predominate 135-kDa protein (p135); however, their ability to stimulate p135 O glycosylation was only consistently observed when islets were isolated in the presence of high glucose and 1 mM L-glutamine. Islets isolated in low glucose and no added L-glutamine demonstrated no consistent increase in p135 O-glycosylation in response to glucose, glucosamine, or streptozotocin. These data suggest that during islet isolation, beta-cell enzymes responsible for regulating p135 O glycosylation may be adversely affected by the absence of high glucose and glutamine, which together are necessary for O-linked N-acetylglucosamine synthesis. We propose that the combination of high glucose and glutamine during islet isolation generates UDP-N-acetylglucosamine and O-linked N acetylglucosamine, thus providing substrate protection for these enzymes and preserving the ability of isolated islets to O-glycosylate p135. PMID- 11019825 TI - Anion-induced folding of rabbit muscle pyruvate kinase: existence of multiple intermediate conformations at low pH. AB - Structural and functional characteristics of rabbit muscle pyruvate kinase (PK), a tetrameric enzyme having identical subunits, were investigated under neutral as well as acidic conditions by using enzymatic activity measurements and a combination of optical methods, such as circular dichroism, fluorescence, and ANS binding. At low pH and low ionic strength, pyruvate kinase exists in a partially unfolded state (UA state) retaining half of the secondary structure and no tertiary interactions along with a strong binding to the hydrophobic dye, ANS. Addition of anions, like NaCl, KCl, and Na2SO4, to the acid-unfolded state induces refolding, resulting structural propensities similar to that of native tetramer. When anion concentration exceeds a critical limit (0.7 M KCl), a sudden loss of secondary structure and decrease in fluorescence intensity with a redshift in the emission maximum are seen which may be due to the aggregation of the protein, probably due to the intermolecular association. The anion-refolded state is more stable than the UA state, and its stability is nearly equal to that of native protein toward chemical-induced unfolding by Gu-HCl and urea. Moreover, at low concentrations, Gu-HCl behaves like an anion, by inducing refolding of the acid-unfolded state with structural features equivalent to that of native molecule. These observations support a model of protein folding where certain conformations of low free energy prevail and are populated under non-native conditions with different stability. PMID- 11019826 TI - Metabolic changes in the poly(ADP-ribosyl)ation pathway of differentiating rat germinal cells. AB - Endogenous levels of poly(ADP-ribose) and betaNAD+ have been determined in rat male germinal cells at different stages of differentiation. The levels of both metabolites decreased progressively from primary spermatocytes to secondary spermatocytes and especially in spermatids. We have also determined the size and complexity of the ADP-ribose polymers synthesized in permeabilized germ cells. Polymers of different chain length and complexity were observed in cells incubated with different concentrations of [32P]betaNAD+; short polymers characterized primary spermatocytes incubated with low betaNAD+ concentration. In all cell fractions, polymers of over 20 residues in size were observed at high betaNAD+ levels. Long polymers were associated with the sulfuric acid-insoluble proteins (nonhistone proteins such as PARP itself). By contrast, oligomers of 20 ADP-ribose units or less were found in the sulfuric acid-soluble proteins (histone proteins). We have also identified the main ADP-ribose protein acceptors formed in each cell type. In all cells examined, PARP appears to be extensively automodified. However, by far, the H1t variant of histone H1 appeared to be the preferred ADP-ribose target among the acid-soluble proteins separated by reverse phase HPLC. Therefore, we conclude that an active protein-poly(ADP-ribosyl)ation system is concentrated in primary spermatocytes, based on a high level of PARP automodification accompanied by the preferential heteromodification of the histone H1 variant specifically expressed in the cells undergoing the pachytene phase of the meiotic division. PMID- 11019827 TI - Intact human ceruloplasmin is required for the incorporation of iron into human ferritin. AB - We have previously reported several studies on the loading of iron into ferritin by ceruloplasmin using proteins from rats. Loading iron into human ferritin using human serum ceruloplasmin is complicated by the fact that human ceruloplasmin is very susceptible to proteolysis (T. P. Ryan, T. A. Grover, and S. D. Aust, 1992, Arch. Biochem Biophys. 293, 1-8). The present study investigated the effect of proteolysis on the ability of human ceruloplasmin to load iron into human ferritin. SDS-PAGE revealed one major band with an apparent molecular weight of 116 kDa for a proteolytically degraded form of ceruloplasmin versus a 132-kDa band for an intact form of the enzyme. Both forms of the enzyme possessed ferroxidase activity, although that of the proteolytically degraded enzyme was approximately twofold less than that of the intact enzyme (4.9 nmol (min)-1 vs 8.3 nmol (min)-1). Only the intact form of ceruloplasmin was able to catalyze iron loading into ferritin without altering the physical characteristics of the ferritin protein during the process. Abnormal migration in nondenaturing PAGE gels, as well as a decrease in the amount of detectable ferritin protein, was observed when ferritin was incubated with iron alone or with proteolytically degraded ceruloplasmin and iron. It was concluded that the structural integrity of ceruloplasmin is required for the enzyme to effectively catalyze iron loading into ferritin. PMID- 11019828 TI - Ethanol has multiple effects on DNA synthesis in fibroblasts depending on the presence of secreted growth regulators and zinc as well as the level of protein kinase C activation. AB - Earlier we showed that in serum-starved (27 h), washed mouse fibroblasts and other cell lines 40-80 mM concentrations of ethanol (EtOH) potentiate, in a zinc (Zn2+)-dependent manner, the combined stimulatory effects of calcium (Ca2+) and insulin (Ins) on DNA synthesis. We now report that the promitogenic EtOH effects require removal of the used medium at least 6 h prior to treatments with EtOH, Zn2+, and Ins. If serum-starved (27 h) cells were continuously incubated for another 18-h period without replacing the medium, a secreted cellular factor moderately enhanced the mitogenic effect of Ins and simultaneously blocked the potentiating effect of EtOH on DNA synthesis measured during the last hour of treatments. However, the presence of Ca2+ (2.8 mM) plus Zn2+ (25 microM) or 25 300 nM phorbol 12-myristate 13-acetate (PMA) during the serum starvation period partially restored the promitogenic effect of EtOH. The PMA effect was blocked by the protein kinase C (PKC) inhibitor GF 109203X added for the second (18 h) period. Even at 300 nM, PMA failed to fully downregulate PKC-alpha, the major PKC isoform, over a 28-h period, suggesting that an activated PKC enzyme was involved in the restoration of EtOH effect. When EtOH (40-80 mM) was added for the entire serum starvation period and the incubations were continued for 18 h without removing the medium, EtOH inhibited both the combined actions of Ins and cellular factor as well as the promoting effect of newly added EtOH on Ins-dependent DNA synthesis. Coaddition of Zn2+ and PMA with EtOH prevented these inhibitory effects of EtOH. The results indicate that in mouse fibroblasts EtOH can both enhance and inhibit Ins-dependent DNA synthesis depending on the timing of EtOH treatment as well as the presence of Zn2+, cellular factors, and activators of the PKC system. PMID- 11019829 TI - Interaction of tetrahydrocrysene ketone with estrogen receptors alpha and beta indicates conformational differences in the receptor subtypes. AB - Estrogen receptors (ER) alpha and beta bind estradiol (E2) and other estrogenic ligands with different affinities. To measure the rate of E2 association with ERa and ERbeta, we employed tetrahydrocrysene ketone (THCK), a fluorescent ligand that is an agonist with ERalpha and an antagonist with ERbeta. We report that THCK binds E2-liganded and unliganded ERalpha and ERbeta, indicating a THCK binding site(s) other than the E2 binding pocket. THCK fluorescence was greater for ligand-occupied ERbeta than ERalpha, suggesting differences in the microenvironment of the THCK binding site(s). THCK fluorescence was also significantly greater for E2-, 4-hydroxytamoxifen-, and tamoxifen aziridine liganded versus unliganded ER, allowing calculations of E2 association rate constants (ka) of 7.60 +/- 0.75 and 5.12 +/- 0.30 x 10(5) M(-1) s(-1) for E2 ERalpha and E2-ERbeta, respectively. THCK did not affect ERalpha binding to estrogen response element (ERE) DNA, but decreased ERbeta-ERE binding. We conclude that THCK binding site(s) on ERalpha versus ERbeta are different and important for ER function. PMID- 11019830 TI - Regulation of 25-hydroxyvitamin D3 1alpha-hydroxylase gene expression by parathyroid hormone and 1,25-dihydroxyvitamin D3. AB - The conversion of 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) takes place mainly in the kidney and is catalyzed by the enzyme 1alpha hydroxylase. Parathyroid hormone (PTH) and 1,25-(OH)2D3 are well-known regulators of this tightly controlled step, but the mechanisms by which they modulate 1alpha hydroxylase activity have not been fully delineated. Northern analysis showed PTH and forskolin rapidly and transiently increase 1alpha-hydroxylase expression in AOK-B50 cells and HKC-8 cells. Actinomycin D treatment blocks the increase, but cycloheximide does not. No decrease of 1alpha-hydroxylase transcript by 1,25 (OH)2D3 was observed in either cell line, although 24-hydroxylase levels were strongly induced by 1,25-(OH)2D3 treatment. 1,25-(OH)2D3 suppressed the 1alpha hydroxylase transcript in vivo both in the presence and absence of exogenously supplied PTH. These results suggest that the stimulatory action of PTH is directly on the 1alpha-hydroxylase gene, while the repressive action of 1,25 (OH)2D3 does not involve the parathyroid gland but is nevertheless indirect. PMID- 11019831 TI - Chemical modification of pyrimidine TFOs: effect on i-motif and triple helix formation. AB - In order to form more stable triple helical structures or to prevent their degradation in cells, oligonucleotide analogs are routinely used, either in the backbone or among the bases. The target sequence chosen for this study is a 16 base-long oligopurine-oligopyrimidine region present in the human neurotrophin 4/5 gene. Seven different chemical modifications were tested for their effect on (i) triple helix formation and (ii) i-DNA stability. i-DNA is a tetrameric structure involving hemiprotonated C x C+ base pairs, which may act as a competing structure for triplex formation, especially in the case of a cytosine rich third strand. At acid pH, oligophosphoramidates formed the most stable triple helix, whereas oligonucleotides including 5-propynyl-dU formed a stable i motif which precluded triplex formation. Only two candidates stabilized triple helices at neutral pH: oligonucleotides with phosphoramidate linkage and phosphodiester oligonucleotides containing 5-methyl-dC and 5-propynyl-dU. PMID- 11019832 TI - Coexpression of genetically engineered fused enzyme between yeast NADPH-P450 reductase and human cytochrome P450 3A4 and human cytochrome b5 in yeast. AB - Human hepatic cytochrome P450 3A4 (CYP3A4) was expressed in yeast Saccharomyces cerevisiae. While the expression level was high as compared with other human hepatic cytochrome P450s, CYP3A4 showed almost no catalytic activity toward testosterone. Coexpression of CYP3A4 with yeast NADPH-P450 reductase did not give a full activity. Low monooxygenase activity of CYP3A4 was attributed to the insufficient reduction of heme iron of CYP3A4 by NADPH-P450 reductase. To enhance the efficiency of electron transfer from NADPH-P450 reductase to CYP3A4, a fused enzyme was constructed between CYP3A4 and yeast NADPH-P450 reductase. The rapid reduction of the heme iron of the fused enzyme by NADPH was observed. The fused enzyme showed a high testosterone 6beta-hydroxylation activity with a sigmoidal velocity saturation curve. However, the coupling efficiency between NADPH utilization and testosterone 6beta-hydroxylation was only 10%. Finally, coexpression of the fused enzyme and human cytochrome b5 was examined. A significant decrease in the Km value and a remarkable increase in the coupling efficiency were observed. Substrate-induced spectra revealed that the dissociation constant of the fused enzyme for testosterone significantly decreased with coexpression of human cytochrome b5. These results strongly suggest that human cytochrome b5 directly interacts with the CYP3A4 domain of the fused enzyme and modifies the tertiary structure of substrate binding pocket, resulting in tight binding of the substrate and high coupling efficiency. PMID- 11019833 TI - Phosphate-substituted ATP analogs are antagonists at human P2Y1 purinoceptors. PMID- 11019834 TI - Individualizing high-dose oral busulfan: prospective dose adjustment in a pediatric population undergoing allogeneic stem cell transplantation for advanced hematologic malignancies. AB - We investigated whether adjusting the oral busulfan (BU) dosage on the basis of early pharmacokinetic data to achieve a targeted drug exposure could reduce transplant-related complications in children with advanced hematologic malignancies. Twenty-five children received a preparative regimen consisting of thiotepa (250 mg/m2 i.v. daily for 3 days), BU (40 mg/m2 per dose p.o. every 6 h for 12 doses), and cyclophosphamide (60 mg/kg i.v. daily for 2 days) and then underwent allogeneic stem cell transplantation. Busulfan clearance and area under concentration time-curve (AUC) were determined after the first dose using a one compartment pharmacokinetic (PK) model with first-order absorption. The initial PK analysis was successfully completed after the first BU dose in 21 patients (84%). A final AUC of 1000-1500 microM x min/dose was targeted and subsequent doses were modified as necessary to achieve this value. Fourteen of the 25 patients (56%) required dose adjustment. Follow-up PK analysis was completed in 21 patients and 16 of these achieved the targeted BU exposure for the course of therapy. Interpatient variability in BU clearance was high (up to five-fold). The most frequent regimen-related toxicities were cutaneous and gastrointestinal (stomatitis and diarrhea). Only one patient developed hepatic veno-occlusive disease. Our study demonstrates the feasibility of adjusting the oral BU dose in individual pediatric patients. Although toxicity associated with BU seemed to be reduced, this conclusion is tempered by the fact that the overall regimen-related toxicity (RRT) remains substantial and reflected the effects of all agents used in the preparative regimen. PMID- 11019836 TI - Docetaxel effectively mobilizes peripheral blood CD34+ cells. AB - We prospectively evaluated docetaxel (100 mg/m2) with G-CSF (10 microg/kg S.C., daily) for mobilization efficiency in 26 patients with breast cancer. The minimum target yield was >4.5 x 10(6) CD34+ cells/kg (optimum = 9 x 10(6)/kg), sufficient to support the subsequent three cycles of high-dose therapy (HDT). The peak days for peripheral blood (PB) CD34+ cells were day 8 and day 9. Seven collections began on day 7, 16 on day 8 and three on day 9. The median peripheral blood progenitor cell (PBPC) CD34+ cell content ranged from 1.2 to 5.9 x 10(6)/kg per day during days 7 to 11 with a median CD34+ content of the total 72 PBPC collections of 3.4 x 10(6)/kg (0.07-15.6). Fifteen patients obtained a PBPC collection exceeding 5 x 10(6)/kg on a single day of collection. Following a median 3 days collection for each patient (range 2-4), the median total CD34+ for all individual sets of collections was 9.7 x 10(6)/kg (range 1.0-28.4). We were able to achieve the minimum CD34+ cell target yield in 22 of 26 patients with one cycle of mobilisation chemotherapy and in two of these patients a second collection yielded sufficient cells. Twenty-two patients have subsequently received repetitive HDT and PBPC transplantation with 57 cycles of HDT having been delivered. For all 57 cycles, the median time to absolute neutrophil count (ANC) >0.5 x 10(9)/l and 1.0 x 10(9)/l was 10 days (range 8-22) and 11 days (range 8-23), respectively. The median time to platelets greater than 20 x 10(9)/l, 50 x 10(9)/l and 100 x 10(9)/l was 13 days (range 11-23), 17 days (range 12-53) and 23 days (range 18-70), respectively. We conclude that docetaxel with G CSF effectively mobilises PBPCs with apheresis needing to be commenced approximately 8 days after docetaxel administration. PMID- 11019835 TI - A randomized phase 2 study of PBPC mobilization by stem cell factor and filgrastim in heavily pretreated patients with Hodgkin's disease or non-Hodgkin's lymphoma. AB - This randomized, controlled study compared the ability to mobilize and collect an optimal target yield of 5 x 10(6) CD34+ cells/kg using stem cell factor (SCF; 20 microg/kg/day) plus filgrastim (G-CSF; 10 microg/kg/day) vs filgrastim alone (10 microg/kg/day) in 102 patients diagnosed with non-Hodgkin's lymphoma (NHL) or Hodgkin's disease (HD), who were prospectively defined as being heavily pretreated. Leukapheresis began on day 5 of cytokine administration and continued daily until the target yield was reached, or until a maximum of five leukaphereses had been performed. Compared with the filgrastim-alone group (n = 54), the SCF plus filgrastim group (n = 48) showed an increase in the proportion of patients reaching the target yield within five leukaphereses (44% vs 17%, P = 0.002); reduction in the number of leukaphereses required to reach the target yield (P = 0.003); reduction in the proportion of patients failing to reach a minimum yield of 1 x 10(6) CD34+ cells/kg to proceed to transplant (16% vs 26%, P = NS); increase in the median yield of CD34+ cells per leukapheresis (0.73 x 10(6)/kg vs 0.48 x 10(6)/kg, P = 0.04); and an increase in the median total CD34+ cells collected within five leukaphereses (3.6 x 10(6)/kg vs 2.4 x 10(6)/kg, P = 0.05). All patients receiving SCF were premedicated (antihistamines and albuterol), and treatment was generally well tolerated. Five patients experienced severe mast cell-mediated reactions, none of which were life-threatening. In this study of heavily pretreated lymphoma patients, SCF plus filgrastim was more effective than filgrastim alone for mobilizing PBPC for harvesting and transplantation after high-dose chemotherapy. PMID- 11019837 TI - A low CD34+ cell dose results in higher mortality and poorer survival after blood or marrow stem cell transplantation from HLA-identical siblings: should 2 x 10(6) CD34+ cells/kg be considered the minimum threshold? AB - We studied the effect of the CD34+ cell dose on transplant-related mortality (TRM) and survival in 39 patients randomized to receive lenograstim-mobilized PBSCT (n = 20) or BMT (n = 19) from HLA-identical siblings. Both marrow and blood were harvested, and one infused in a double-blind fashion. The median nucleated (7.0 vs 3.2 x 10(8)/kg; P < 0.0001), CD34+ (3.7 vs 1.5 x 10(6)/kg; P = 0.002), CFU-GM (42 vs 19 x 10(4)/kg; P = 0.002), and CD3+ (1.9 vs 0.3 x 10(8)/kg; P < 0.0001) cell doses with PBSCT were higher. Thirteen patients (6 BMT and 7 PBSCT) experienced TRM at 15-733 days (median 57); 10 of 20 receiving <2 x 10(6) CD34+ cells/kg compared with three of 19 receiving > or =2. Eight of 20 patients receiving <2 x 10(6) CD34+ cells/kg are alive compared with 14 of 19 receiving > or =2. In Cox analysis, CD34+ cell dose > or =2 x 10(6)/kg was associated with lower TRM (RR 0.2, P = 0.01), and higher overall (RR 3.7, P = 0.01) and event free (RR 3.2, P = 0.02) survival. Other cell populations and the source of stem cells did not affect TRM or survival. We conclude that 2 x 10(6) CD34+ cells/kg may be the ideal minimum cell dose for allogeneic transplantation although lower doses do not preclude successful therapy. Since the likelihood of obtaining this threshold CD34+ cell number is significantly greater from blood than marrow, PBSCT may be preferable to marrow for allografts from HLA-identical siblings. PMID- 11019838 TI - Ex vivo expansion of CD34-positive peripheral blood progenitor cells from patients with non-Hodgkin's lymphoma: no evidence of concomitant expansion of contaminating bcl2/JH-positive lymphoma cells. AB - The aim of the present study was to evaluate the capacity to expand of hematopoietic stem cell (HSC) samples from eight patients with NHL, and to follow in parallel the fate of tumor cells in four of eight samples still containing bcl2/JH+ tumor cells after CD34+ or CD19-/20-/34+ cell selection. The presence of bcl2/JH+ cells was also investigated after expansion in four of eight samples, two of which were bcl2/JH at harvesting and two which were initially bcl2/JH+ but became bcl2/JH (below the level of PCR detection) after cell selection, to assess a possible reappearance of occult tumor cells after expansion culture. We used culture conditions that we previously had established to allow high level expansion of normal precursors, progenitors and LTC-ICs. In this study, particular attention was given to the role of Flt3-ligand, known to favor the growth of B cells. The expansion conditions were: 1.5 x 10(3) cells/ml in serum free medium containing stem cell factor (SCF), interleukin-3 (IL-3), IL-6, granulocyte-stimulating factor (G-CSF), erythropoietin (Epo) +/- Flt3-ligand (Flt3-L) for 10 days. After culture, total cells, CFU-GMs, BFU-Es and LTC-ICs were expanded to a mean of 833-, 6.6-, 4.6-, and 1.8-fold, respectively with the cocktail of cytokines not including Flt3-L. When Flt3-L was added, the mean expansion values were 1095-, 31-, 15- and three-fold, respectively. Residual bcl2/JH+ cells present in four of eight samples before expansion were not detected after expansion. Similarly, no tumor cells reappeared after expansion of the two samples which had become negative after selection, as well as in the two samples which were bcl2/JH- at harvesting. These results suggest first that ex vivo expansion of hematopoietic stem cells in patients with non-Hodgkin's lymphoma is feasible without incurring the parallel risk of amplifying tumor cells; second, that Flt3-L did not stimulate the growth of tumor cells while it clearly favored the growth of normal progenitors. PMID- 11019839 TI - Clinical impact of ex vivo differentiated myeloid precursors after high-dose chemotherapy and peripheral blood progenitor cell rescue. AB - The infusion of ex vivo differentiated myeloid precursors may be able to shorten the period of obligatory neutropenia after high-dose chemotherapy and peripheral blood progenitor cell rescue by providing cells capable of differentiating to mature neutrophils within days of infusion. To test this hypothesis, 21 female patients with metastatic breast cancer underwent progenitor cell mobilization with cyclophosphamide, etoposide and G-CSF. CD34+ cells from one to two leukapheresis products were isolated and placed in suspension culture with a serum-free growth medium supplemented with PIXY321. The cultures were maintained for 12 days with subcultures initiated on day 7. The remaining leukapheresis products were cryopreserved in an unmanipulated state. Forty-eight hours after completing high-dose cyclophosphamide, thiotepa and carboplatin, the cryopreserved progenitors were infused, followed 1 to 24 h later by infusion of the differentiated myeloid precursors. In one patient, the cultured cells were labeled with Indium-111 with nuclear imaging performed up to 48 h post infusion. The differentiated myeloid precursors were suitable for infusion in 17 of the patients with a median 13-fold expansion of total nucleated cells. A range of 5.6 to 1066 x 10(7) nucleated cells were infused. Morphologically the cells were predominantly of myeloid lineage (63%) with a median 41% of the cells expressing CD15. No untoward effects were noted with the infusion of the cultured cells. The median days to neutrophil and platelet recovery were 8 and 10 days, respectively. There was a significant relationship (r = 0.67, P = 0.007) between the dose of differentiated myeloid precursors (CD15+ cells) and the depth and duration of neutropenia; a similar relationship, however, was also observed with the dose of cryopreserved CD34+ cells. After infusion of the radiolabeled myeloid precursors, a pattern of distribution similar to radio-labeled granulocytes was noted with uptake detected initially in the lungs and subsequently the reticulo-endothelial system. The impact of differentiated myeloid precursors on neutropenia as an adjunct to high-dose chemotherapy and peripheral blood progenitor cell rescue remains unclear from this study. Further study with controlled doses of cryopreserved progenitors and escalating doses of differentiated myeloid precursors is required. PMID- 11019840 TI - Donor leukocyte infusions in acute lymphocytic leukemia. AB - Donor leukocyte infusion (DLI) has well-documented activity in CML but the role of DLI in other diseases is less well defined. To evaluate the strategy in acute lymphocytic leukemia (ALL) we evaluated 44 ALL patients from 27 centers who were treated with DLI. Patients with persistent or recurrent disease received DLI from the original marrow donor (30 matched related, four mismatched family, and 10 matched unrelated). Chemotherapy was given before DLI to 28 patients. Of 15 patients who received no pre-DLI chemotherapy, two achieved complete remissions, lasting 1112 and 764+ days. In four patients who received DLI as consolidation of remission induced by chemotherapy or immunosuppression-withdrawal, duration of remission post DLI was 65, 99, 195 and 672+ days. Of 25 patients who received DLI in the nadir after chemotherapy, 13 survived > or =30 days post DLI but did not achieve remission, seven died within less than 30 days post DLI, and five entered remissions that lasted 42, 68, 83, 90, 193 days. Seven patients who did not respond to the initial DLI received a second DLI; none of these patients attained durable remission. Eighteen of 37 evaluable patients developed acute GVHD and five of 20 evaluable patients developed chronic GVHD. Overall actuarial survival is 13% at 3 years. In conclusion, DLI has limited benefit in ALL. New approaches are needed in this group of patients. PMID- 11019841 TI - Cytotoxic T lymphocyte precursor frequency as a predictor of acute graft-versus host disease in bone marrow transplantation from HLA-identical siblings. AB - The measurement of precursor frequencies of donor anti-recipient cytotoxic T lymphocytes (CTL-p) has been shown to predict the incidence and the severity of acute graft-versus-host disease (aGVHD) in unrelated donor bone marrow transplantation (BMT). In HLA-identical sibling BMT, where aGVHD is most likely caused by minor histocompatibility antigen mismatches, this assay did not appear to be sensitive enough to provide similar predictive information. In this study, the CTL-p frequencies and the incidence and severity of aGVHD in 51 onco hematological patients transplanted from HLA-identical siblings were compared. Sibling donors were selected on the basis of HLA identity using serological typing for HLA-A, B, C antigens, whereas HLA-DRB was tested by molecular analysis. Sibling identity was also confirmed by DNA heteroduplex analyses. Fifteen out of 21 (71%) patients with high precursor frequency (>1:100 x 10(3)) and 12 out of 30 (40%) with low precursor frequency (<1:100 x 10(3)) experienced clinically significant (II-IV) aGVHD. A significant correlation (P = 0.04) between CTL-p frequency and severe aGVHD was demonstrated. Moreover there was a positive trend for a high frequency response according to an increasing grade of aGVHD, which was statistically significant (P = 0.04). In our experience the CTL p assay is a helpful predictive test for aGVHD in HLA-identical sibling BMT, indicating high risk patients possibly requiring additional prophylaxis. PMID- 11019842 TI - Flow cytometric detection of growth factor receptors in autografts and analysis of growth factor concentrations in autologous stem cell transplantation: possible significance for platelet recovery. AB - In order to improve prediction of hematopoietic recovery, we conducted a pilot study, analyzing the significance of growth factor receptor expression in autografts as well as endogenous growth factor levels in blood before, during and after stem cell transplantation. Three early acting (stem cell factor (SCF), Flt3 ligand (Flt3) and fetal antigen 1 (FA1)) and three lineage-specific growth factors (EPO, G-CSF and thrombopoietin (Tpo)) were analyzed by ELISA in 16 patients with multiple myeloma (MM) and 16 patients with non-Hodgkin's lymphoma (NHL). The relative number of SCF, Flt3, Tpo and G-CSF receptor positive, CD34+ progenitor cells were measured by flow cytometry in the leukapheresis product used for transplantation in a subgroup of 15 patients (NHL, n = 8, MM, n = 7). Three factors were identified as having a significant impact on platelet recovery. First, the level of Tpo in blood at the time of the nadir (day +7). Second, the percentage of re-infused thrombopoietin receptor positive progenitors and finally, the percentage of Flt3 receptor positive progenitors. On the other hand, none of the analyzed factors significantly predicted myeloid or erythroid recovery. These findings need to be confirmed in prospectively designed studies. PMID- 11019843 TI - Utility of obtaining blood cultures in febrile neutropenic patients undergoing bone marrow transplantation. AB - Infection remains an important cause of morbidity and mortality after bone marrow or stem cell transplantation. To evaluate the role of obtaining blood cultures for intermittent or persistent fever in neutropenic patients on antibiotic therapy, we performed a retrospective chart review of 196 consecutive patients admitted to the Bone Marrow Transplant Unit at the University of North Carolina Hospitals from 1995 to 1998. From the cohort of 196 patients, 154 patients developed neutropenic fever. The initial blood culture was positive in 16 of 145 patients during the first fever episode giving a prevalence of 11%. From the total of 109 patients that had blood cultures drawn after day 1 of fever, five patients had blood cultures positive for a pathogen, a prevalence of 4.6%. In only one patient, did blood cultures drawn after day 1 identify an organism not present on day 1 (prevalence 0.9%). After reviewing the results in the first 105 patients, we changed our timing of collection of blood cultures. Forty-nine patients were treated in this manner and we found that the mean number of blood cultures decreased from 9.2 to 4.7 per patient without a change in the frequency of infectious complications or length of hospitalization. PMID- 11019844 TI - Increasing incidence of diffuse alveolar hemorrhage following allogeneic bone marrow transplantation: cryptic etiology and uncertain therapy. AB - Diffuse alveolar hemorrhage (DAH) is a non-infectious pulmonary complication of bone marrow transplantation (BMT) with resultant high mortality. It reportedly occurs primarily in autologous recipients. We examined the incidence of DAH in our center in order to assess potential risk factors and develop preventive strategies. Between 1991 and 1997, 23 cases of DAH occurred in 922 adult patients (2.5%) receiving BMT for hematological malignancy. Strikingly, 12 cases occurred in 1997 with the majority in recipients of allogeneic matched sibling donor stem cells. Treatment with high-dose steroids, 250 mg to 2 g/day, in 15 patients led to transient improvement in 10 patients, but 21 of the 23 patients required mechanical ventilation. Mortality was high with 17 patients (74%) dying a median of 39 days (range 22-47) post transplant; a median of 17 days post onset of DAH (range 5-34). Six patients are alive with a median follow-up of 18 months (range 12-60). No recognizable alteration in supportive care, conditioning regimen, GVHD prophylaxis or cytokine usage was associated with this striking increase in the frequency of DAH after allografting. Further follow-up is required to establish whether this increase in the incidence of DAH in allogeneic transplantation is an isolated occurrence or an ongoing problem. If indeed there is a real increase in the incidence of this complication, then efforts need to be directed towards elucidating a possible cause or risk factors. We offer the possibility that a new unidentified infection, undetected by current microbiological tests might contribute to this striking increase in DAH. These data, while not establishing a cause, suggest a markedly augmented risk of DAH in allogeneic BMT. In addition, high-dose corticosteroids have only limited efficacy as therapy for DAH after allotransplantation. Further investigation into the pathogenesis of this syndrome is essential as is prompt and immediate consideration of DAH in all patients with respiratory compromise early after BMT. PMID- 11019845 TI - Effect of gastrointestinal inflammation and age on the pharmacokinetics of oral microemulsion cyclosporin A in the first month after bone marrow transplantation. AB - Cyclosporin A (CsA) absorption is highly variable in BMT patients. Neoral, a new microemulsion formulation of CsA, permits increased absorption with less variable pharmacokinetic parameters in non-BMT patients. We evaluated the pharmacokinetics of CsA after BMT in patients received microemulsion CsA. Two oral doses of 3mg/kg were given 48 h apart between 14 and 28 days after allogeneic BMT in 20 adults, and one dose in seven children, while subjects were receiving a continuous i.v. infusion of CsA. Whole blood samples were taken throughout the dosing interval to calculate the incremental CsA exposure using maximum concentration (Cmax), time to Cmax (tmax), concentration at 12 h after the dose (C12), the area under the concentration-time curve (AUC), and to establish inter- and intra-patient pharmacokinetic variability. Drug exposure was substantially lower in children than adults, with an AUC of 861+/-805 vs 2629+/-1487 micromg x h/l (P = 0.001), respectively, and absorption was delayed and diminished in both groups by comparison with solid organ recipients. Intra-patient variability in adults for AUC was high at 0.59+/-0.34, while inter-patient variability, measured as the coefficient of variation (c.v.), was 0.55 for the first and 0.54 for the second dose. In adults, gastrointestinal (GI) inflammation due to either mucositis or GVHD resulted in a higher AUC of 3077+/-1551 microg x h/l compared to 1795+/-973 microg x h/l (P = 0.02), and a similar trend was observed in children. AUC seemed little affected by the CsA formulation (liquid or capsule), or co-administration with liquids or food. Trough (12 h) CsA levels correlated poorly with incremental AUC. Sparse sample modeling of the AUC using two-point predictors taken at 2.5 and 5 h after dosing accurately approximated AUC in adults (r2 = 0.94), while 1.5 and 5 h was superior in children (r2 = 0.98). These data suggest that 12 h postdose trough measurements of CsA may not be the most appropriate way to evaluate CsA blood concentrations in order to establish therapeutic efficacy in BMT patients. Based on this study, the dose of microemulsion CsA should be adjusted based on recipient age, and the presence of GI inflammation secondary to mucositis or GVHD. These data would suggest that sparse sampling at time points earlier than the trough more accurately reflects the AUC and may correlate more closely with therapeutic efficacy early post-BMT. PMID- 11019846 TI - Predictors of physical outcomes in pediatric bone marrow transplantation. AB - The purpose of the present study was to investigate the hypothesis that family factors, in conjunction with clinical factors, are associated with physical outcomes in pediatric BMT. A prospective study of 68 pediatric patients (mean age = 7.5 years; ranging from 4 months to 18 years) undergoing BMT was carried out over a 6.5 year period. Physicians rated initial prognosis on a (0-5) scale which incorporated the child's diagnosis, known risk factors, and type of donor. Both parents individually completed two psychometrically sound questionnaires assessing family well-being and marital satisfaction. Cox proportional hazards survival analyses were performed to determine predictors of death (44% of the patients died). Potential predictor variables included were: initial prognosis, type of transplant, patient's age, socioeconomic status, marital satisfaction and family status, and family stress. Initial prognosis, as estimated by the physician, (RR = 0.62, 95% CI = 0.40, 0.97) was the best predictor of survival. Initial clinical factors are clearly critical in outcomes for pediatric BMT patients. PMID- 11019847 TI - Kinetics of in vivo homing and recruitment into cycle of hematopoietic cells are organ-specific but CD44-independent. AB - In this study, we investigated the homing and initiation of division of fluorescently labelled adult mouse bone marrow cells after their intravenous injection into lethally irradiated congenic mice. After 2 h, only 3% of the transplanted cells remained in the blood, and approximately 35% could be retrieved from the marrow, liver and spleen in approximately equal numbers. Subsequently, the proportion of injected cells found in blood, liver and spleen decreased further, but increased slightly (to approximately 17%) in the marrow. Homing of progenitors followed a similar pattern. At 22 h post transplant, almost half of the injected cells in the blood, liver and spleen had completed a first mitosis; although these did not include progenitors with in vitro clonogenic ability. At the same time, >90% of the injected cells recovered from the marrow had not yet divided. Parallel studies with CD44-/- mice showed these to contain a numerically and functionally normal stem cell population whose homing and activation in either CD44+/+ or CD44-/- hosts appeared unaltered. These results indicate homing mechanisms that favor more stable retention of transplanted marrow cells in the marrow of the recipient, more rapid activation of some of those cells that home to other sites, and a lack of change in either of these responses when either the transplanted or the recipient cells do not express CD44. PMID- 11019848 TI - Wolman disease successfully treated by bone marrow transplantation. AB - Wolman disease is characterized by severe diarrhea and malnutrition leading to death during infancy. Lysosomal acid lipase deficiency is the cause of the symptoms and signs. It is inherited in an autosomal recessive manner. All Wolman disease patients have adrenal gland calcification. Previous therapeutic attempts have failed to provide remission. We report successful long-term bone marrow engraftment in a patient with Wolman disease resulting in continued normalization of peripheral leukocyte lysosomal acid lipase enzyme activity. Diarrhea is no longer present. Now, at 4 years of age, this patient is gaining developmental milestones. Cholesterol and triglyceride levels are normal. Liver function is normal. This is the first long-term continued remission reported for Wolman disease. PMID- 11019849 TI - Peripheral stem cell transplantation in a child with amegakaryocytic thrombocytopenia. AB - Congenital amegakaryocytic thrombocytopenia (CAMT) is an unusual cause of thrombocytopenia without radial or other congenital anomalies in the newborn. Generalized bone marrow dysfunction developing later in life has been reported. We present a 13-month-old girl who was diagnosed as having congenital amegakaryocytic thrombocytopenia and was successfully treated with allogeneic peripheral stem cell transplantation (PSCT) from her fully matched sibling donor. The neutrophil engraftment was on post transplant day 12 and platelet engraftment was on day 14. Her last hemogram revealed platelets of 168 x 10(9)/l 20 months post transplant. PMID- 11019850 TI - Donor lymphocyte infusion for treatment of life-threatening respiratory syncytial virus infection following bone marrow transplantation. AB - We describe two patients who developed respiratory syncytial virus (RSV) pneumonia after BMT. One died of RSV pneumonia after three courses of steroid pulse therapy. Surprisingly, RSV antigen was identified in the bronchoalveolar lavage fluid (BALF) obtained post mortem. Steroid pulse therapy might have suppressed anti-RSV immunity, leading to persistent RSV infection for more than 1 month. The other patient received donor lymphocyte infusions (DLI) for relapsed plasma cell leukemia, while having active RSV pneumonia. His respiratory condition improved after DLI, and RSV antigen disappeared in BALF and nasal swabs. DLI might be effective in cases of life-threatening RSV pneumonia. PMID- 11019851 TI - Epstein-Barr virus-negative high grade B cell lymphoma of donor origin developing 19 months after unrelated allogeneic bone marrow transplantation. AB - A 22-year-old man, in first complete remission of acute myelogenous leukemia, developed a high grade B cell lymphoma 19 months after an allogeneic bone marrow transplant (allo-BMT) from an HLA-identical unrelated donor. Biopsy of a cervical lymph node revealed a lymphoma that was negative for Epstein-Barr virus-encoded small nuclear RNAs (EBERs) in situ hybridization. Genotypic analyses identified the lymphoma to be of donor origin, and there was no evidence of the Epstein-Barr virus (EBV) DNA in the lymphoma by Southern blot analysis. The lymphoma went into complete remission, following four courses of combination chemotherapy, but relapsed after a month and the patient died of congestive heart failure. The patient was thought to be persistently immunosuppressed 11 months after cessation of immunosuppressants, and the lymphoma was thought to be induced by one or more factors other than EBV. PMID- 11019852 TI - Histologic transformation of follicular lymphoma after allogeneic bone marrow transplantation. AB - A 38-year-old man with refractory follicular lymphoma underwent allogeneic BMT from an HLA-identical sibling donor. He had generalized lymphadenopathy, hepatosplenomegaly and lymphoma infiltration of the marrow, all of which disappeared within 3 months following transplantation. Six months post transplant, progressive hepatomegaly developed in the absence of splenomegaly and lymphadenopathy, and he died from hepatic failure. Autopsy disclosed diffuse large B cell lymphoma of the liver, into which the follicular lymphoma had transformed. Future issues to be investigated should include the optimal timing of allogeneic BMT for low-grade lymphomas. PMID- 11019853 TI - Searching for a cord blood unit for transplantation based on the unit's cell count. AB - Although recipients of unrelated cord blood can tolerate a greater degree of HLA disparity than recipients of unrelated bone marrow, cell dose is also important. After an unsuccessful search for a bone marrow donor, and after initially dismissing a cord blood search because the patient was 52 kg, we used a different search strategy to identify an unrelated cord blood unit. This led to successful engraftment and transplantation for a patient with no bone marrow donor. PMID- 11019854 TI - Early viral complications after autologous CD34-selected peripheral blood stem cell transplantation. PMID- 11019855 TI - Autologous PBSCT in patients with cardiac amyloidosis. PMID- 11019856 TI - Beta amyloid fragments derived from activated platelets deposit in cerebrovascular endothelium: usage of a novel blood brain barrier endothelial cell model system. AB - Amyloid precursor protein (A betaPP) processing results in generation of amyloid beta peptide (A beta) which deposits in the brain parenchyma and cerebrovasculature of patients with Alzheimer's disease (AD). Evidence that the vascular deposits derive in part from A betaPP fragments originating from activated platelets includes findings that individuals who have had multiple small strokes have a higher prevalence of AD compared to individuals who have taken anti-platelet drugs. Thus, determination of whether platelet A betaPP fragments are capable of traversing the blood-brain barrier (BBB) is critical. We have established that activated platelets from patients with AD retain more surface transmembrane-bound A betaPP (mA betaPP) than control platelets. We report here that this mA betaPP can be cleaved to A beta-containing fragments which pass through a novel BBB model system. This model utilizes human BBB endothelial cells (BEC) isolated from brains of patients with AD. These BEC, after exposure to activated platelets which have been surface-labeled with fluorescein and express surface-retained mA betaPP, cleave fluorescein-tagged surface proteins, including mA betaPP, resulting in passage to the BEC layer The data confirm that BEC contribute to processing of platelet-derived mA betaPP and show that the processing yields A beta containing fragments which could potentially contribute to cerebrovascular A beta deposition. PMID- 11019857 TI - An improved method of preparing the amyloid beta-protein for fibrillogenesis and neurotoxicity experiments. AB - Synthetic amyloid beta-protein (A beta) is used widely to study fibril formation and the physiologic effects of low molecular weight and fibrillar forms of the peptide on cells in culture or in experimental animals. Not infrequently, conflicting results have arisen in these studies, in part due to variation in the starting conformation and assembly state of A beta. To avoid these problems, we sought a simple, reliable means of preparing A beta for experimental use. We found that solvation of synthetic peptide with sodium hydroxide (A beta x NaOH), followed by lyophilization, produced stocks with superior solubility and fibrillogenesis characteristics. Solubilization of the pretreated material with neutral buffers resulted in a pH transition from approximately 10.5 to neutral, avoiding the isoelectric point of A beta (pI approximately 5.5), at which A beta precipitation and aggregation propensity are maximal. Relative to trifluoroacetate (A beta x TFA) or hydrochloric acid (A beta x HCl) salts of A beta, yields of "low molecular weight A beta" (monomers and/or dimers) were improved significantly by NaOH pretreatment. Time-dependent changes in circular dichroism spectra and Congo red dye-binding showed that A beta x NaOH formed fibrils more readily than did the other A beta preparations and that these fibrils were equally neurotoxic. NaOH pretreatment thus offers advantages for the preparation of A beta for biophysical and physiologic studies. PMID- 11019858 TI - Histidine residues underlie Congo red binding to A beta analogs. AB - The binding mechanism of Congo red (CR) to Alzheimer's disease (AD) amyloid fibrils (A beta) in terms of binding affinity and number of sites was quantitated from absorption spectroscopy (at 200-700 nm) by measuring the concentration of CR bound (CR-B) to AD A beta assemblies as a function of CR concentration and pH in 80% ethanol. The rationale for the use of this high concentration of ethanol derives from its use in histological screens for amyloid in tissue sections. Moreover, free CR can be separated from bound CR by filtration in ethanolic but not aqueous medium. The A beta analogs studied here included: (1) peptides having different lengths: A beta1-40, A beta11-28, A beta13-28, A beta19-28, A beta11 25; (2) wildtype, control sequences of A beta1-40 and sequences having different natural amino acid substitutions: primate Pr1-40, rodent Ro1-40, hereditary cerebral haemorrhage with amyloidosis, Dutch type (HCHWA-D) Du1-40, primate reverse sequence Pr40-1; and (3) A beta11-25 sequences having different substitutions: H13D, H14D, and D23K. Negative-staining showed that A beta1-40 fibrils in buffer were indistinguishable from those in buffered ethanolic medium. For all amyloid analogs except A beta19-28, which has no histidine residues and showed no CR binding over the entire pH range 4.0-9.5, CR-B decreased as a function of increasing pH. The decrease was steepest at about pH 5 and became zero above pH 7. For analogs having the same number of histidines, CR-B fell on the same binding curve, indicating that histidine residues are the likely binding sites for CR in this medium. The pH titration of the binding was parameterized by the stoichiometry of dye to the sites, the number of histidines per molecule, the binding dissociation constant Kd, and the apparent proton dissociation constant pK of the histidine; and the calculated pH-titration curves were found to fit the observed ones. For the peptides having 1-3 histidines the average pK was 5.0-5.5, which was similar to the expected pK of histidine in low dielectric medium (80% ethanol), and the Kd's were 2.8-5.9 microM. That histidine residues underlie CR binding in A beta amyloid is consistent with previous findings that A beta peptides sediment as fibrillar assemblies at pH-3-7 and bind Congo red over the same pH range in aqueous medium. Further, the conformation near the binding motif His13-His14-Gln15-Lys16 in A beta assemblies is not greatly altered in 80% ethanol. PMID- 11019859 TI - Transformation of beta-amyloid (A beta) (1-42) tyrosine to L-dopa as the result of in vitro hydroxyl radical attack. AB - A form of beta-amyloid peptide A beta ending at amino acid 42 (A beta42) is the major component of senile amyloid plaques in Alzheimer's Disease (AD). The A beta peptide earliest modifications are extremely important since they constitute the key events in the progression towards further changes finally leading to fibril formation and to A beta deposits which constitute the core pathological change in AD. Chemical and conformational early modifications of the beta-amyloid peptide are critical steps in AD pathogenesis and have been widely investigated. We now show that a Fenton-type OH-generating system is capable of generating L-Dopa (3,4 dihydroxyphenylalanine) in the tyrosine residue of A beta-peptide via aromatic ring hydroxylation, as the result of hydroxyl radical attack on proteins. Since L Dopa is not a constituent of mammalian proteins and peptides, the formation of L Dopa in A beta in vitro constitutes a possible important modification caused by hydroxyl radical attack. These results lay the groundwork for further studies on modification and damage associated with the degenerative disorder in AD where oxidative stress and inflammation are known to occur. PMID- 11019860 TI - Amyloid peptide channels: blockade by zinc and inhibition by Congo red (amyloid channel block). AB - Amyloid peptides are the major constituents of amyloid deposits in various amyloid diseases including Alzheimer's disease, type II diabetes mellitus, prion diseases and others. The hallmark of amyloid is the binding of the dye, Congo red, which creates characteristic staining due to the dye's ability to bind the beta sheet aggregates referred to as amyloid. Previous reports have demonstrated that several cytotoxic, amyloidogenic peptides can form ion channels in planar phospholipid bilayer membranes and have suggested that these channels may represent the pathogenic mechanism of cell and tissue destruction in amyloid disease. Furthermore, zinc and Congo red can ameliorate or prevent the pathogenic effect of certain amyloidpeptides. We report here that zinc at micromolar concentrations caused a reversible blockade of islet amyloid polypeptide (IAPP, amylin) and PrP 106-126 channels whereas calcium and magnesium did not. Congo red completely inhibited channel formation if preincubated with amyloid peptides, but had no effect on IAPP or PrP 106-126 channels once formed. These results suggest a requirement for aggregation for the formation of amyloid peptide channels and are consistent with the "channel hypothesis" of amyloid disease. They also suggest potential avenues for ameliorative therapy of these illnesses. PMID- 11019861 TI - Four structural risk factors identify most fibril-forming kappa light chains. AB - Antibody light chains (LCs) comprise the most structurally diverse family of proteins involved in amyloidosis. Many antibody LCs incorporate structural features that impair their stability and solubility, leading to their assembly into fibrils and to their subsequent pathological deposition when produced in excess during multiple myeloma and primary amyloidosis. The particular amino acid variations in antibody LCs that account for fibril formation and amyloidogenesis have not been identified. This study focuses on amyloidogenesis within the kappa1 family of human LCs. Reanalysis of the current database of primary structures of proteins from more than 100 patients who produced kappa1 LCs, 37 of which were amyloidogenic, reveals apparent structural features that may contribute to amyloidosis. These features include loss of conserved residues or the gain of particular residues through mutation at sites involving a repertoire of approximately 20% of the amino acid positions in the light chain variable domain (V(L)). Moreover 80% of all kappa1 amyloidogenic V(L)s are identifiable by the presence of at least one of three single-site substitutions or the acquisition of an N-linked glycosylation site through mutations. These findings suggest that it is feasible to predict fibril propensity by analysis of primary structure. PMID- 11019862 TI - Histochemical and immunohistochemical differential diagnosis of amyloidosis--a brief illustrated essay and personal experience with Romhanyi's method. AB - The histochemical and immunohistochemical differential diagnosis of amyloidosis in surgical pathology in a referral center is presented. Different forms of amyloidosis are considered e.g. systemic generalized amyloidosis: secondary (AA), primary (AL), senile, hemodialysis-associated, hereditary and organ (tissue) limited (localized) amyloidosis: cerebral, dystrophic (age-related, so-called "senile"), endocrine-related, localized to tumours, focal (concentrated secretion), and isolated plasma cell (solitary plasmacytoma, B-cell) dyscrasia related amyloidosis. The amyloid deposits were identified and characterized histochemically by Congo red staining after performate pre-treatment at 20 degrees C for 1, 3, 5, 10, 15, 20 or 25 sec, and with oxidation induced proteolysis by trypsin digestion at 20 degrees C for 5, 10, or 30 sec, 1, 2, 3, 4, 5, 6 or 10 min and covered with gum-arabic according to Romhanyi, and confirmed by streptavidin-biotin-complex/horseradish peroxidase immunohistochemical reactions. The "sensitivity" or "resistance" to pre-treatment of amyloid deposits depends on the type of amyloid, and the length of pre treatment. Secondary (AA) amyloid is sensitive to KMnO4 oxidation, followed by trypsin digestion (for 1 min), and its green birefringence under polarized light disappears, while primary (AL) (for 1-5 min), senile (for 1-10 min), and most forms of organ (tissue)-limited (localized) amyloid (for 1-10 min) are resistant. Performate pre-treatment is followed by pronounced congophilia. Secondary (AA) is sensitive to performate pre-treatment (for 1 sec), while primary (AL) amyloid (for 1-20 sec), senile (for 1-25 sec), and most forms of organ (tissue)-limited (localized, isolated) amyloid deposits (for 1-25 sec) are resistant, and are constantly positively birefringent. Early identification and differentiation of amyloid deposits is important for the prognosis and for the choice of therapy. The authors conclude that the presented classical histochemical methods are useful as first line screens for the histological identification of amyloidosis. PMID- 11019863 TI - Transthyretin Val71Ala mutation in a Dutch family with familial amyloidotic polyneuropathy. AB - A Dutch family with familial amyloidotic polyneuropathy associated with the transthyretin mutation Val71Ala is described. This is the third reported family with this mutation, causing at the protein level an unstable TTR monomer and at the clinical level progressive wasting, polyneuropathy, autonomic dysfunction and vitreous opacities. PMID- 11019865 TI - Surgically-induced astigmatism after laser in situ keratomileusis for spherical myopia. AB - PURPOSE: To characterize the surgically-induced-astigmatism (SIA) associated with spherical LASIK. METHODS: Refractive outcomes in 70 eyes that underwent primary myopic LASIK with purely spherical ablation were analyzed. The Summit Apex Plus excimer laser was used. The Bausch & Lomb Hansatome with the 180-microm plate was used to produce superiorly hinged flaps. The relationship between refractive astigmatism and corneal topographic astigmatism was analyzed using linear regression and vector analysis. RESULTS: There was a statistically significant negative correlation (slope = -0.21) between refractive surgically-induced astigmatism and preoperative topographic cylinder. A 0.24-D with-the-rule shift was also found. Surgically-induced astigmatism was not correlated with the magnitude of laser ablation. CONCLUSION: The lamellar keratotomy portion of LASIK reduces pre-existing corneal astigmatism and produces a relative steepening of the hinge meridian. PMID- 11019864 TI - Corneal first surface optical aberrations and visual performance. AB - PURPOSE: Wavefront analysis has demonstrated that refractive surgery-induced corneal first surface aberrations are large, are dominated by symmetric aberrations (spherical-like aberrations), and are correlated to measures of visual performance. It is not clear whether the correlation between corneal first surface aberrations and visual performance can be generalized to other corneal conditions where large asymmetric aberrations (coma-like aberrations) may dominate the aberration structure. The purpose of the research reported here was to determine the general utility of corneal first surface wavefront analysis in predicting visual performance. METHODS: Patients were 13 normals and 78 patients with a variety of corneal conditions including surgically removed pterygia, penetrating keratoplasty, keratoconus, radial keratotomy, laser in situ keratomileusis, and others. Videokeratographs were taken for all patients and used to calculate corneal first surface wavefront variance for 3 and 7 mm pupils. Similarly, visual performance was quantified by measurements of contrast sensitivity and high and low contrast acuities through both 3 and 7 mm pupils. RESULTS: Statistically significant correlations existed between all three measures of visual performance and the corneal wavefront variance. All relationships were stronger for the 7 mm diameter-pupil condition than the 3 mm pupil. CONCLUSION: Regardless of the cause, corneas with increased wavefront variance showed a quantifiable decrease in visual performance that was pupil size dependent. PMID- 11019866 TI - Pain after epithelial removal by ethanol-assisted mechanical versus transepithelial excimer laser debridement. AB - PURPOSE: To compare subjective pain responses between two techniques of epithelial removal prior to photorefractive keratectomy (PRK) treatment: ethanol soaked pledget with mechanical debridement of the epithelium versus excimer laser transepithelial ablation. METHODS: Nine patients underwent bilateral PRK. Each had the epithelium in one eye debrided by placing a pledget soaked in 20% ethanol on the cornea for 2 minutes followed by gentle scraping with a blade. The epithelium in the other eye was removed by transepithelial phototherapeutic keratectomy (PTK) treatment. For each eye, PRK was initiated immediately after removal of the epithelium. On postoperative day one, each patient was asked to rate the level of pain suffered over the last 24 hours on a scale of 0 (minimal) to 10 (maximal). Data were analyzed in a masked fashion. RESULTS: Postoperative day one average pain level in the ethanol-assisted mechanically debrided eyes was 3.0 +/- 2.5 and in the transepithelial PTK eyes was 6.8 +/- 1.8. The difference was statistically significant by Student's t-test (P < .01). All epithelial defects healed within 3 days with no clinically significant difference in healing time between the two techniques. CONCLUSION: In preparation for PRK, ethanol assisted mechanical debridement of the epithelium caused significantly less postoperative pain than epithelial removal using the excimer laser. PMID- 11019867 TI - Postoperative inflammation, microbial complications, and wound healing following laser in situ keratomileusis. AB - Although the biology of corneal wound healing is only partly understood, healing after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) differs in many respects, and the mechanisms appear to be differently controlled. There is less of an inflammatory and healing response after LASIK, but a longer period of sensory denervation. The cellular, molecular, and neural regulatory phenomena associated with postoperative inflammation and wound healing are likely to be involved in the adverse effects after LASIK, such as flap melt, epithelial ingrowth, and regression. Interface opacities in the early postoperative period include diffuse lamellar keratitis (DLK), microbial keratitis, epithelial cells, and interface opacities. Diffuse lamellar keratitis (sands of the Sahara syndrome) describes an apparently noninfectious diffuse interface inflammation after lamellar corneal surgery probably caused by an allergic or a toxic inflammatory reaction. Noninfectious keratitis must be distinguished from microbial keratitis to avoid aggressive management and treatment with antimicrobial drugs. Microbial keratitis is a serious complication after LASIK, but a good visual outcome can be achieved following prompt and appropriate treatment. PMID- 11019868 TI - The prospects for perfect vision. PMID- 11019869 TI - Limits to vision: can we do better than nature? AB - Non-invasive wavefront sensing of the human eye provides the necessary information to design corrections which minimize the monochromatic optical errors of the eye beyond simple sphere (defocus) and cylinder (astigmatism). These "ideal" corrections must move with the eye, maintaining proper alignment with the eye's optics. Viable modes of correction include contact lenses, refractive surgery and intraocular lenses. Will these "ideal" corrections lead to better vision? If so, how much better? Here we explore the limits imposed by the optical and neural design of the eye. For larger pupil sizes (>3 mm diameter) "ideal" corrections improve the optical quality of the retinal image beyond the limits imposed by photoreceptor spacing. Photoreceptor spacing limits visual acuity to between 20/8 and 20/10. Correcting the higher order aberrations will provide images with higher contrast and crisper edges. When perfected, "ideal" corrections will provide for high contrast visual acuity between 20/8 and 20/10. PMID- 11019870 TI - The history and methods of ophthalmic wavefront sensing. PMID- 11019871 TI - Visual benefit of correcting higher order aberrations of the eye. AB - There is currently considerable debate concerning the visual impact of correcting the higher order aberrations of the eye. We describe new measurements of a large population of human eyes and compute the visual benefit of correcting higher order aberrations. We also describe the increase in contrast sensitivity when higher order aberrations are corrected with an adaptive optics system. All these results suggest that many, though not all, observers with normal vision would receive worthwhile improvements in spatial vision from customized vision correction, at least over a range of viewing distances and particularly when the pupils are large. Keratoconic patients or patients suffering from spherical aberration as a result of laser refractive surgery as it is presently performed would especially benefit. These results encourage the development of methods to correct higher order aberrations. PMID- 11019872 TI - Understanding aberrations by using double-pass techniques. PMID- 11019873 TI - Principles of Hartmann-Shack aberrometry. PMID- 11019875 TI - Principles of Tscherning aberrometry. AB - Higher-order optical errors of the human eye are often responsible for reduced visual acuity in spite of an optimal spherical or cylindrical refraction. These optical aberrations are of natural origin or can result from operations on the eye involving optical structures. The presented wavefront analyzer is based on Tscherning's aberroscope. A collimated laser beam illuminates a mask with regular matrix pin holes which forms a bundle of thin parallel rays. These rays form a retinal spot pattern on the retina that is more or less distorted according to the optical errors of the eye. This retinal spot pattern is imaged onto the sensor of a low-light CCD camera by indirect ophthalmoscopy. The deviations of all spots from their ideal regular positions are measured by means of a personal computer, and from these values the optical aberrations are computed in the form of Zernike polynomials up to the 8th order. PMID- 11019874 TI - The spatially resolved refractometer. AB - PURPOSE: To investigate factors controlling the aberrations of the eye, including accommodation, wavelength, and the apodization of the optics of the eye by cone directional selectivity. METHODS: We constructed a new implementation of the Spatially Resolved Refractometer (SRR). This is an instrument, based on the Scheiner principle, that allows the rapid psychophysical measurement of the complete wavefront aberrations of the eye. We have investigated both the reproducibility of the measurements, and the effect of static accommodation and wavelength on the wavefront aberrations of the eye. In addition we combined the wave front aberrations with cone photoreceptor directionality to compute the modulation transfer function of the eye, at the retinal level. RESULTS: The SRR measurements were rapid (4 minutes per measurements, 12 minutes per patient) and reproducible. There are significant changes in wavefront quality with accommodation, with optimal image quality near the resting point of accommodation. Image quality for polychromatic (white) light is strongly affected by both longitudinal and transverse chromatic aberration. Finally, we find that incorporating the effects of cone directionality into the calculation of image quality can increase image quality by up to 50%. CONCLUSION: Calculation of a simple "optimal surgical shape" for wave-front guided refractive surgery will depend on improved understanding of the interplay between the biological and physical properties of the eye. PMID- 11019876 TI - Principles of ray tracing aberrometry. AB - PURPOSE: Of all transforms of an eye, aberrations are significant when higher visual acuity is to be achieved. Ray tracing aberrometry developed by the Institute of Biomedical Engineering (Kiev) and first tested at the Vardinoyannion Eye Institute of Crete is a promising technique for eye refraction aberration and refraction mapping. METHODS: The technique uses measurement of the position of a thin laser beam projected onto the retina. The beam is directed into the eye parallel to the visual axis. Each entrance point provides its own projection on the retina. A set of entrance points forms a set of projections. From these data, a refraction map is reconstructed as well as a point spread function of the eye. The total time of scanning over the whole aperture of the eye is within 10 to 20 ms and depends on the number of test points at the eye entrance, as well as on the number of independent measurements in each point. Configuration of the scanning pattern can be chosen by the operator. It may contain 60 to 400 points, each checked 1 to 5 times. RESULTS: Preliminary studies showed high reproducibility of results. Twenty pseudophakic eyes were subjected to 30 consecutive measurements each. Ninety-five percent of all measured values were within +/-0.20 D of declination from the mean. CONCLUSIONS: Ray tracing aberrometry is a flexible technology for eye investigation. It can be adapted to any laser technique of vision correction Its further development should be oriented on laser-linked applications of the refraction driven refractive surgery. PMID- 11019877 TI - Slit skiascopic-guided ablation using the Nidek laser. AB - PURPOSE: To present the approach of using a scanning slit refractometer (the ARK 10000) in conjunction with a corneal topography system to guide customized corneal ablation. This diagnostic system is coupled with the Nidek EC-5000 system which combines scanning slit and a scanning small area ablation (1.0 mm) to perform a customized ablation. METHODS: The ARK 10000 diagnostic system which contains a scanning slit refractometer is described. Information generated from the ARK 10000 wavefront sensor and corneal topography system can be coupled to the new Nidek EC-5000 excimer laser system, which combines the larger area of scanning slit ablation with the small area (1.0 mm) ablation. RESULTS: The Nidek ARK 10000 diagnostic system captures wavefront information using a retinoscopic system which is converted into a refractive power map. This is different from other autorefraction systems in that it has four sensors at different diameters of the cornea and captures 1440 points in 0.4 seconds. This map is used in conjunction with corneal topography-captured simultaneously. This information is then combined to perform a customized ablation using the new Nidek EC-5000 system. CONCLUSIONS: The ARK 10000 diagnostic system represents a different approach to customized ablation in that it combines a corneal topography system with a wavefront system and a larger treatment area of the traditional scanning slit ablation with a new small area ablation treatment for greater efficiency. PMID- 11019878 TI - Clinical experience with the CustomCornea measurement device. PMID- 11019879 TI - Clinical experience with the Tscherning aberrometer. AB - PURPOSE: With the aberrometer based on Tscherning's principle, measurements of wavefront aberrations of human eyes with high accuracy and reproducibility are available for standard diagnostic investigations. METHODS: During investigational and clinical trials, wavefront-aberrations of about 300 human eyes were measured and evaluated within the last few years. RESULTS: measurements are presented in terms of Zernike coefficients and as height maps that can be converted directly to ablation profiles for wavefront-guided laser treatments. CONCLUSION: The Tscherning aberrometer is a simple optical device with high accuracy appropriate for routine clinical investigations on optical aberrations of the human eye. PMID- 11019880 TI - Clinical experience with the Tracey technology wavefront device. AB - PURPOSE: In refractive surgery, measuring the total ocular profile of refraction gives more vital information than measuring the cornea alone. We report the first clinical experiments with spatially resolved refractometry using ray tracking principles. Tracey technology was developed in Ukraine by the Institute of Biomedical Engineering in cooperation with the Vardinoyannion Eye Institute of Crete. METHODS: The Tracey-1 device was evaluated in comparison with conventional videokeratoscopes. Seven pseudophakic eyes of 7 patients and two phakic eyes were measured thirty (30) consecutive times (consecutively) each to test reproducibility. Measurements were provided in zones 0 to 5 (6) mm. For each zone, parameters of astigmatism were calculated and mean value and standard deviation were derived. RESULTS: Standard deviation on the order of 0.14 D was derived. CONCLUSION: The Tracey Technology wavefront device provides information on the refraction distribution at the first principal plane of the ocular optical system. For the instrument to provide measurements that are directly applicable to clinical practice, the data should be transposed to the corneal plane. The Tracey device can be utilized for the measurement of accommodative amplitude and range. Further development of the instrument is requred to increase its accuracy. PMID- 11019881 TI - Technology requirements for Summit-Autonomous CustomCornea. PMID- 11019882 TI - Adaptive optics ophthalmoscopy. AB - Retinal images in the human eye are normally degraded because we are forced to use the optical system of the human eye--which is fraught with aberrations--as the objective lens. The recent application of adaptive optics technology to measure and compensate for these aberrations has produced retinal images in human eyes with unprecedented resolution. The adaptive optics ophthalmoscope is used to take pictures of photoreceptors and capillaries and to study spectral and angular tuning properties of individual photoreceptors. Application of adaptive optics technology for ophthalmoscopy promises continued progress toward understanding the basic properties of the living human retina and also for clinical applications. PMID- 11019883 TI - Preoperative simulation of outcomes using adaptive optics. AB - Measurements of the wavefront of light reflected from the retina of the human eye can be used to determine optical aberrations of the human eye for large pupils. An instrument based on the Hartmann-Shack principle was developed. The wavefront is refracted by a microlens array and detected by a CCD camera. In first clinical studies human volunteer eyes and preoperative and postoperative refractive surgical patient eyes have been examined. An adaptive optical closed loop system has been devised for preoperative simulation of refractive outcomes of aberration free refractive surgical procedures. PMID- 11019884 TI - Modeling and predicting visual outcomes with VOL-3D. PMID- 11019885 TI - Summit-Autonomous CustomCornea laser in situ keratomileusis outcomes. PMID- 11019886 TI - Operative correction of ocular aberrations to improve visual acuity. AB - PURPOSE: Optical aberrations of the human eye degrade the quality of the retinal image and may, therefore, represent a major limit of visual acuity. METHODS: In 15 eyes, ocular aberrations were corrected in addition to myopia and astigmatism by means of wavefront-guided laser in situ keratomileusis (LASIK). RESULTS: At 1 month after surgery, a supernormal visual acuity of 20/10 and better was obtained in 4 eyes (27%). The increase in root mean square wavefront error ranged from 0.6 to 2.3 and was significantly correlated with the increase in visual acuity (R2 = 0.79; P = .03). CONCLUSION: Although the correction of aberrations was not yet optimal, these results show that ocular optical aberrations limit visual acuity in humans and supernormal visual acuity can be achieved by operative correction. PMID- 11019887 TI - Future challenges to aberration-free ablative procedures. PMID- 11019888 TI - Line of sight and alternative representations of aberrations of the eye. AB - Several methods for representing pupil plane aberrations based on wavefront height, slope, and curvature are discussed. The choice of reference axis and reference surface is shown to strongly affect the appearance of the representation. Special attention will be paid to the use of the line of sight (the line from the fixation point to the center of the pupil) as the reference axis. We show that the line of sight is ambiguous and does not determine the amount of prism in the wavefront. PMID- 11019889 TI - A quick method for analyzing Hartmann-Shack patterns: application to refractive surgery. AB - A quick method to analyze the Hartmann-Shack pattern was developed. This method allows us to study the limitations of the system and to determine how to improve its capacity to measure a large range of aberration. It can treat situations presenting difficulty when analyzing a detected pattern. An application to eyes with large amounts of aberration due to corneal surgery is considered. PMID- 11019890 TI - Determining the accuracy of an eye tracking system for laser refractive surgery. AB - PURPOSE: Patient eye and head movements during laser refractive surgery may result in errors between the surgical beam position and the desired location for optimum correction. This, in turn, may lead to reduced postoperative vision, including increased higher order aberrations of the eye. Active eye tracking systems are often incorporated into laser delivery systems, which aim to reduce the effect of patient eye movement. METHODS: In this study, the accuracy of an eye tracking system designed for laser refractive surgery was determined. An enucleated porcine eye was attached to a scanning device and the movement measured using the eye tracking system. The recorded position is compared to the preprogrammed position of the scanning device. RESULTS: The system demonstrated an accuracy of 0.06 m for an intact cornea and 0.1 mm for a cornea with a thin flap removed. This compares to an average decentration of ablation of 0.4 mm for patients relying on passive fixation, as measured by previous clinical trials. CONCLUSION: implementation of this eye tracker would lead to improved alignment between the laser and eye during laser refractive surgery. PMID- 11019891 TI - Longitudinal evaluation of optical aberrations following laser in situ keratomileusis surgery. AB - Objective measurements of the optical aberrations of an eye were taken with a Shack-Hartmann aberrometer immediately before, immediately after, and at regular intervals over a two month recovery period following LASIK refractive surgery. Results indicate that the surgery induced large amounts of positive spherical aberration which regressed slightly during recovery. Computed point-spread functions for various pupil diameters indicate that retinal image quality was the same before and after surgery for small and medium sized pupils (<4 mm diameter) but was significantly degraded for large pupils (>4 mm). PMID- 11019892 TI - New paradigm for the treatment of myopia by refractive surgery. AB - It is proposed that refractive surgery visual outcomes will be significantly improved when using refractive surgery profiles that maintain preoperative corneal aberrations following surgery. An equation to calculate postoperative corneal shapes is presented. PMID- 11019893 TI - Report from the VSIA taskforce on standards for reporting optical aberrations of the eye. PMID- 11019894 TI - Reference axis selection: subcommittee report of the OSA Working Group to establish standards for measurement and reporting of optical aberrations of the eye. AB - It is the committee's recommendation that the ophthalmic community use the line of sight as the reference axis for the purposes of calculating and measuring the ocular optical aberrations. PMID- 11019895 TI - High resolution MRI study of circumlental space in the aging eye. PMID- 11019896 TI - Measurement of corneal thickness by videopachymetry: preliminary results. AB - Corneal thickness measurements find applications in areas such as diagnosis and management of corneal disorders and corneal surgery. We present a technique for the measurement of corneal thickness using a CCD camera mounted on a slit-lamp biomicroscope and common image processing software. Thickness measurements were performed on three RGP contact lenses of known thickness with an average error of 5 microm, SD of 8 microm. Measurements of a living cornea yielded a thickness of 505 microm and a SD of 8 microm for 10 consecutive measurements; this is an acceptable value. PMID- 11019897 TI - Phakic IOLs revisited; the current FDA trials. PMID- 11019899 TI - Suckling-induced change in oxytocin but not in alpha-MSH concentrations of the median eminence, the neural-, intermediate- and anterior lobes of the pituitary gland. AB - Previous reports have implicated that pituitary-derived prolactin (PRL) is secreted from two distinct zones of mammotropes within the anterior lobe (AL). The inner zone (AL-IZ), located adjacent to the neuro-intermediate lobe (NIL), is supposed to be involved in the rapid and massive discharge of PRL from the pituitary gland due to suckling stimulus. Whereas the outer-zone (AL-OZ) gives the basal secretion and it does not play a role in the acute secretory response during nursing. Anatomically, the AL-IZ has an intimate contact with the NIL because the blood passing through the short portal vessels (SPV) bathes it first. Based on this fact it would be hypothesized that locally released and/or produced compounds, like OXY and alpha-MSH, can be delivered to the AL-IZ. In conjunction, OXY and alpha-MSH have already been implicated to play a role in the regulation of PRL release during suckling. Therefore, the purpose of this study was to examine the possible local transportation of these hormones into the median eminence and various regions of the pituitary gland of lactating rats. We have measured the concentrations of OXY and alpha-MSH from tissue samples of nonsuckled (NS) and 10 or 30 min after suckling (S) was initiated using specific RIAs. It has been shown that there are no changes in the concentration of OXY and alpha-MSH in theAL-IZ and AL-OZ due to suckling stimulus. In contrast, our data provide compelling evidence that OXY is transported into the IL, which can be further increased by suckling stimulus. These data suggest that blood transfusing NL passes through the IL before it is drained into the cavernous sinus, which opens the road for OXY into the general circulation. In addition, our data have unequivocally shown a lack of local delivery of either alpha MSH or OXY into the AL that raises serious doubt about their possible role in PRL secretion during suckling stimulus. PMID- 11019898 TI - Tyrphostin-23 enhances steroid-hormone secretion from dispersed human and rat adrenocrotical cells. AB - Tyrphostin-23 is commonly used as inhibitor of tyrosine kinase (TK). We found that tyrphostin-23 concentration-dependently increased basal steroid-hormone secretion from dispersed human and rat adrenocortical cells, the maximal effective concentration being 10(-5) M. Tyrphostin-23 (10(-5) M) enhanced 10(-9) M angiotensin-II- and endothelin-1-stimulated secretion of human and rat adrenocortical cells, but not the secretory response to 10(-9) M ACTH However, it increased the response to lower concentrations (10(-12) or 10(-11) M) of ACTH. The secretagogue effect of tyrphostin-23 on dispersed rat adrenocortical cells was abolished by either the adenylate cyclase inhibitor SQ-22536 (10(-4) M) or the protein kinase A (PKA) inhibitor H-89 (10(-5) M). Tyrphostin-23 (10(-5) M) raised basal cyclic-AMP release by dispersed rat adrenocortical cells, but in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX, 10(-3) M) it was ineffective. Both tyrphostin-23 and IBMX increased cyclic-AMP release by rat adrenocortical cells in response to 10(-10) M ACTH, and their effects were not additive. Taken together, our findings suggest that tyrphostin 23, acting as an inhibitor of phosphodiesterases in adrenocortical cells, increases the intracellular concentration of cyclic-AMP available for PKA activation thereby stimulating steroid-hormone secretion. They also stress that caution must be used in interpreting the results of studies aimed at investigating the possible cross-talk between adenylate cyclase- and TK-dependent signaling cascades. PMID- 11019900 TI - Involvement of endogenous corticotropin-releasing factor in mediation of neuroendocrine and behavioral effects to alpha-melanocyte-stimulating hormone. AB - The present work was to study if the alpha-melanocyte-stimulating hormone (alpha MSH) was involved in activation of the pituitary-adrenal axis (PAA) in rats. The hormone increased plasma corticosterone (CORT) level, and induced an anxiogenic response as indicated by results from the elevated plus-maze test. Intracerebroventricular administration of corticotropin-releasing factor (CRF) antiserum (1:10, 1:20 and 1:100 dilutions in 1microl volume), overcame both the anxiogenic response and the PAA activating effect induced by alpha-MSH (50 microg s.c.) in a concentration-dependent manner. CRF antibody at the doses applied did not modify either the elevated plus-maze responses or CORT level by itself. Our results reveal that both the anxiogenic and the PAA activating effects of alpha MSH are mediated by CRF. PMID- 11019901 TI - Human galanin secretion is increased upon normal exercise test in middle-age individuals. AB - Galanin, a neuropeptide, is found in the central nervous system and in a number of nonbrain areas including adrenal sympathetic medullar tissue and pancreas. Several studies involve galanin in the regulation of GH, which responds to stressful stimuli. This study refers to the investigation of the effect of a 20 min exercise on plasma human galanin (hGAL) and GH in middle-aged healthy volunteer adults. Thirteen individuals, 5 males and 8 females aged 40-50 years (44.7 +/- 2.95) were selected on the basis of normal body mass index (22.5 +/- 2.3 kg/m2) and the absence of endocrine or any other abnormality. Basal concentrations of GH and hGAL were measured between 0800 and 0900h after an overnight fast. Post exercise levels were recorded after termination of the stressful test and 15 min thereafter. GH and hGAL were determined by an immunoradiometric and radioimmunoassay, respectively. The exercise-potentiated GH response in all subjects with post-exercise levels significantly higher (11.09 +/ 1.8 ng/ml vs 1.27 +/- 0.7 ng/ml, p<0.0001, F=32.44) with the peak in the hormone level detected 15 min after the end of exercise (12.09 +/- 1.96 ng/ml). Plasma hGAL levels were also substantially affected by the acute exercise test, in that post exercise peripheral blood concentration was significantly higher from the basal values (21.51 +/- 9.94 vs 13.46 +/- 7.2 pg/ml, p<0.02, F=5.50). Again the hGAL values peaked 15 min after the end of exercise (24.0 +/- 10.5 pg/ml, P<0.015, F = 4.68). However, the time-correlation of the increments of GH and hGAL did not reach a statistically significant level (20 min: r=0.41, p=0.161., 35 min: r=-0.095, P=0.758). These results clearly show an independence of the two hormones. The responsivity of hGAL of middle-aged individuals to the exercise stimulus might be due to the higher releasable pool of the hormone. PMID- 11019902 TI - Oxidative protein damage in type I diabetic patients with and without complications. AB - To examine the influence of oxidative stress on oxidative protein damage, we studied 47 Type I diabetic patients with and without complications. We determined plasma protein carbonyl, plasma protein thiol and nitrotyrosine levels as markers of oxidative protein damage, plasma lipid hydroperoxide levels as markers of oxidative stress, and plasma total thiol, plasma nonprotein thiol, erythrocyte glutathione, plasma ceruloplasmin, transferrin and total iron binding capacity as markers of free radical scavenging. There were no significant differences in nitrotyrosine, total plasma thiol, protein thiol, and erythrocyte glutathione levels between diabetic patients with complications and without complications. However, plasma protein carbonyl, lipid hydroperoxide, and nonprotein thiol levels were significantly increased in diabetic patients with complications compared with diabetic patients without complications. Although redox status of plasma is impaired in diabetic patients, we suppose these significantly different markers reflect enhanced oxidative protein damage in diabetic patients with complications. PMID- 11019903 TI - The effect of short-term treatment with micronized estradiol on bone turnover and gonadotrophins in older men. AB - Evidence for the role of estrogen in male bone metabolism has been confirmed by studies on a man with a genetic defect in the estrogen receptor as well as men with aromatase defects. All exhibited tall stature, delayed epiphysial closure, decreased bone density and increased bone turnover. Estrogen is likely to affect bone turnover in men throughout life; therefore, we hypothesized that older men would show decreased bone resorption in response to estrogen therapy. To test our hypothesis, fourteen community-dwelling men with osteopenia of the femoral neck were treated for 9 weeks with micronized estradiol, 1 mg/d, a dose which is effective in postmenopausal women. Each subject served as his own control. Markers of bone resorption, N-terminal collagen crosslinks (NTX) and C-terminal collagen crosslinks (CTX) and markers of bone formation, osteocalcin (OC) and bone specific alkaline phosphatase (BSAP) were measured every 3 weeks during a 9 week treatment period and 9 weeks post-treatment. Sex hormones, gonadotrophins and calciotropic hormones were measured at baseline, 9 weeks on treatment and 9 weeks post- treatment. After 9 weeks of treatment, estradiol and estrone levels increased significantly by greater than 6-fold and 15-fold, respectively. SHBG levels increased significantly by 17%. Testosterone and free testosterone levels decreased significantly by 27% and 34%, respectively. Markers of bone resorption showed wide variation at baseline and while on treatment. There was no correlation between changes in bone markers and changes in estrogen levels. During treatment, 11 patients showed a decrease of NTX or CTX, but three showed an increase. These three and one other subject had high initial levels of FSH and LH, suggesting some degree of primary gonadal failure, which decreased during estrogen therapy. Thus, the change in NTX (and CTX) after 9 weeks of E2 treatment was correlated with initial FSH (r= -.66, p= .01) and LH (r= -.73, p= .003) values. In addition, the largest decrease in free testosterone at 9 weeks was correlated with the higher values for NTX, CTX and BAP (r=-0.66, -0.68, -0.70 respectively; p< or =.01 for each of the markers). Treatment was generally well tolerated. Side effects of treatment were minimal, and included breast tenderness and decreased libido which reversed after treatment. We conclude that it is feasible to give low dose estrogen to healthy older men, but that the effects on bone turnover are not consistent. Changes in central feedback and in endogenous sex hormone production may alter the response of bone turnover to exogenous estrogen in this population. PMID- 11019904 TI - Interrelationships of GABAergic, serotoninergic and excitatory amino acid systems in its regulatory effect on prolactin secretion in prepubertal rats. AB - GABAergic, serotoninergic and excitatory amino acid systems (EAAs) regulate the prolactin (PROL) secretion in prepubertal female rats. The aim of the present paper was to determine the interrelationships of these systems on the control of this pituitary hormone. It was carried out through the following scheme: 1. The participation of the EAAs and serotonin in the effect of GABAergic system on PROL release, determined by evaluating the GABA A and GABA B receptor agonists. It was carried out on animals that were previously treated with AAEs receptor antagonist or p-chlorophenylamphetamine (PCA), this one depleting serotonin in the hypothalamus. 2. The participation of GABAergic system in the effect of serotonin and EAAs systems, determined by the evaluation of the effects of EAAs receptor agonists and of 5-HTP, a serotonin precursor. With this purpose the rats were previously treated with GABA A and GABA B receptor antagonists. 3. The interrelationships between the EAAs and the serotoninergic systems in the control of PROL secretion, determined (a) by using EAAs agonists (in rats depleted of serotonin by PCA) and (b) using EAAs antagonists (in rats treated with 5-HTP, a serotonin precursor). The administration of GABAergic agonists significantly increased PROL secretion in prepubertal female rats. Neither EAAs antagonists nor the depletion of serotonin in the brain, modified the stimulatory effects of the GABAergic system on PROL levels. This is a clear indication that the activity of the GABAergic system is independent of the serotoninergic and of the EAAs system effects on the pituitary hormone. The EAAs neurotransmitter system agonists significantly increase PROL levels. This effect was blocked by the GABAergic system antagonists but was not modified by serotonin depletion. Taking into account these facts it may be considered that the GABAergic system is involved in the stimulatory effect of EAAs on PROL secretion, this effect being independent of the serotoninergic system. 5-HTP significantly increased PROL plasma levels, and this effect was modified neither by the GABAergic nor by the EAAs receptor antagonists. These results indicate that the stimulatory effect of serotonin on PROL release is independent of the GABAergic and EAAs systems. In conclusion it may be considered that in prepubertal female rats, the GABAergic and serotoninergic systems stimulate PROL secretion by independent mechanisms that do not include EAAs. On the other hand, the effects of EAAs neurotransmission are exerted via the GABAergic system. PMID- 11019905 TI - Transient neonatal hypothyroidism alters plasma and testicular sex steroid concentration in puberal rats. AB - The stimulatory and inhibitory effects on testicular steroidogenesis of transient neonatal hypothyroidism from day 1 postpartum through different postnatal developmental events on testis at puberal age (60 days old) were studied in vivo. Hypothyroidism was induced in neonates by feeding the lactating mother or directly with 0.05% methimazole (MMI) through drinking water from the day of parturition to 10, 15, 30, 40 and 60 days, and were killed at day 60 postpartum. Plasma and testicular interstitial fluid (TIF) progesterone, testosterone, dihydrotestosterone (DHT) and estradiol concentrations were assessed. Testis weight and volume significantly increased in rats subjected to 10 and 15 days of hypothyroidism, decreased in rats subjected to 30, 40 and 60 days of hypothyroidism. A consistent increase in Leydig cell number was seen in puberal rats subjected to transient neonatal hypothyroidism but decreased in 60 days hypothyroid rats. Peritubular myoid cell number was consistently decreased in all experimental rats. Leydig cell diameter decreased consistently in all experimental groups. Persistent hypothyroidism (60 days hypothyroid) consistently decreased both plasma and TIF sex steroids. In transient hypothyroid rats, progesterone concentration decreased in both plasma and TIF. Transient hypothyroidism from birth to day 10 postnatal age maintained normal titre of plasma testosterone, whereas a significant increase in TIF testosterone concentration was evident when compared with controls. All other groups of rats subjected to transient neonatal hypothyroidism had consistently low titres of plasma and TIF testosterone. Plasma DHT concentrations in rats subjected to transient neonatal hypothyroidism remained unaltered. However, TIF DHT increased in 10 days PMID- 11019906 TI - 3,5,3'-triiodothyronine (T3) and 3',5'-diiodothyrone (T2) have short-term effects on lipid metabolism in a teleost Anabas testudineus (Bloch): evidence from enzyme activities. AB - As thyroid hormones are known to have long-term and short-term effects on metabolism and only the long term effects been examined in detail, in the present investigation a comparative study has been mad to elucidate the short-term effects of T3 and T2 on enzymes of lipid metabolism in a fish Anabas testudineus. The time dependent studies involved assays of specific, activities of lipogenic enzymes and membrane ATPase- Anabas responded to T3 treatment with a significant increase in the liver malic enzyme activity (after 1 hr) and the activity pattern was reversed after 6 hrs in low dose (0.25microg T3) treated group. T2 treatment also increased the malic enzyme activity within one hour after administration. T2 caused an Increase, in the activity of glucose-6-phosphate dehydrogenase under both time courses, HMG-CoA Reductase activity was reduced upon T3 and T2 administration except for T2 at one hour. T3 treatment significantly enhanced Na+K+-ATPase activity while T2 decreased it except by low dose at one hour. Both T3 and T2 treatment influenced the activities of enzymes of lipid synthesis in a dose and time dependent manner emphasising a short-term effect of these thyroid hormones in Anabas. PMID- 11019907 TI - Interactions of CRH, AVP and cortisol in the secretion of ACTH from perifused equine anterior pituitary cells: "permissive" roles for cortisol and CRH. AB - To further elucidate the interaction of CRH, AVP and cortisol in the control of ACTH secretion, we used an in vitro perifusion model with dispersed equine anterior pituitary cells. To approximate the in vivo milieu in the horse, CRH was perifused continuously (at 0, 2 and 20 pmol/L) and 5-min pulses of AVP (0, 1, 3 and 10 nmol/L) were given every 30 min in the presence of 0 or 100 nmol/L cortisol. Total (baseline + incremental) ACTH secretion increased as both the CRH (p<0.001) and the AVP (p<0.001) concentration increased and interaction between CRH and AVP was significant (p=0.042). Cortisol reduced total ACTH secretion in the presence of 2 pmol CRH/L (p=0.001) but not 0 or 20 pmol CRH/L. For incremental ACTH there was interaction between CRH and AVP (p<0.0001), with increased secretion at higher concentrations, and no significant main effect of cortisol. There was significant (p=0.001) interaction between cortisol and CRH, with cortisol attenuating ACTH release at 0 pmol CRH/L (p=0.008), having no effect at 2 pmol CRH/L and potentiating it at 20 pmol CRH/L (p=0.026). We conclude that (1) CRH at high physiological levels has a "permissive" role in preventing the cortisol inhibition of the ACTH response to AVP, and (2) basal cortisol levels have a "permissive" action in priming the HPA axis for maximal responsiveness to stimulated levels of CRH and AVP. PMID- 11019908 TI - Plasma acyl-estrone levels are altered in obese women. AB - A group of obese women (BMI>27 kg/m2; N=73) was studied together with lean controls (BMI <27 kg/m2; N=25). Three groups were defined by the compliance with: BMI lower than 27 kg/m2, glycaemia lower than 5.5 mM and insulinaemia lower than 0.2 nM (controls, group 1, N=19). The subjects with BMI>27 kg/m2, glucose >5.5 mM and insulin >0.2 nM constituted group 3 (N=41), and those with BMI>27 with glycaemia and/or insulinaemia lower than the limits set constituted group 2 (N=32). The women in group 3 had higher fat content, BMI and fat-free mass than those in group 2 and the controls. There were no changes in most plasma parameters, such as free estrone and beta-estradiol. Leptin levels were higher in groups 2 and 3 than in controls. In controls, leptin and acyl-estrone levels were well correlated with BMI and fat content; this correlation was not found in groups 2 and 3 for acyl-estrone, although it was found for leptin. Acyl-estrone levels were lower than expected in most obese women when compared to those of controls, suggesting an altered availability or function of this hormone. In obese women, acyl-estrone levels -and probably function- are lower than expected, contrasting with maintained leptin-BMI correlations. The role of insulin in the control of body weight, perhaps through acyl estrone-mediated effects, should be re-evaluated. PMID- 11019909 TI - Identification of functional somatostatin receptors and G-proteins in a new line of human foetal lung fibroblasts. AB - A new line (FP) of human foetal lung fibroblasts was analysed for the expression of functional, G-protein coupled somatostatin receptors (SSTR). By means of RT PCR, we identified the expression of SSTR1, SSTR2, SSTR3 and SSTR4, but not SSTR5, subtypes. The same technical approach evidenced the expression of stimulatory (alphas) and inhibitory (alphai1, alphai2 and alphai3) G-protein subunits. The functionality of SSTR was established from the observation of a dose-dependent inhibitory role of SST upon isoproterenol-stimulated adenylyl cyclase activity, an effect that involves G-protein action. Moreover, the functionality of G-proteins was assessed by means of experiments with forskolin and a nonhydrolysable GTP analogue that showed either Gi or Gs activation in the regulation of adenylyl cyclase. Present results represent a first pharmacological characterization of this new line of human foetal lung fibroblasts. The selective presence of some SSTR subtypes and G-protein subunits in addition to the regulatory network of the adenylyl cyclase pathway are features of recognized involvement in cell growth mechanisms. It is of interest for a cell class widely used to study this topic but also important in lung physiology and pathophysiology. PMID- 11019910 TI - Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is associated with a naive B-cell phenotype in human tonsils. AB - In B cells, signaling through the B-cell antigen receptor (BCR) is negatively modulated by the co-ligation of immunoglobulin (Ig)-immunoreceptor tyrosine-based inhibitory motif (ITIM)-bearing molecules such as FcgammaRIIB1, B-cell transmembrane protein CD72, paired immunoglobulin-like receptor PIR-B, leukocyte associated immunoglobulin-like receptor-1 (LAIR-1), Ig-like transcript ILT2, biliary glycoprotein BGP-1 and B-cell co-receptor CD22. The co-expression of multiple Ig-ITIM receptors may provide B cells with different mechanisms of regulating inhibitory pathways at different stages of differentiation. In this study, we have examined the expression of a newly defined Ig-ITIM receptor, PECAM 1 (CD31) on human B-cells. Human tonsillar B cells were purified using negative selection by depleting T cells with a combination of monoclonal antibodies and magnetic bead separation. Following purification, the pattern of PECAM-1 expression was analyzed in B-cell subpopulations using two- and three-colour fluorescence. To complement this work, PECAM-1 localization in the context of distinct areas of human tonsil was defined by immunohistochemical analysis of tonsil sections. Finally to investigate somatic mutation, Ig variable (V) region genes belonging to the nonpolymorphic VH6 family were amplified by polymerase chain reaction (PCR), subcloned and sequenced from sort-purified CD19+ PECAM-1+ and CD19+ PECAM-1- B cells. Our results demonstrate that PECAM-1 is associated with an unstimulated resting B-cell phenotype, localization to the follicular mantle and marginal zones of human tonsil and expression of unmutated Ig V region genes. These studies suggest that PECAM-1 appears on the cell surface at the naive B-cell stage and is lost as B cells differentiate into memory cells, indicating that PECAM-1 is primarily involved in naive or immature B-cell function. PMID- 11019911 TI - Production and characterization of monoclonal antibodies against conserved epitopes of P-selectin (CD62P). AB - P-selectin (CD62P) is an adhesion molecule expressed on the activated endothelium and activated platelets that is involved in the initial attachment of leukocytes to inflamed vascular endothelium. Blocking monoclonal antibodies (mAbs) and P selectin-deficient mice have shown that P-selectin is a potential target in anti inflammatory therapy. Most mAbs against P-selectin do not bind to conserved epitopes, including the ligand-binding region, since P-selectin from mammalian species shares high amino acid sequence homology. The aim of this study was to generate a novel panel of anti-P-selectin mAbs against the conserved epitopes present in several animal species. To produce these mAbs, P-selectin-deficient mice were immunized with a pre-B-cell line transfected with human P-selectin cDNA. Twelve mouse mAbs that recognize human P-selectin were obtained. Individual mAbs that bound to human, rat, mouse, rabbit and pig activated platelets were characterized by flow-cytometry, immunohistochemistry, adhesion assays and immunoprecipitation. Four of these mAbs (P-sel.KO.2.3, P-sel.KO.2.4, P-sel.KO.2.7 and P-sel.KO.2.12) cross-reacted with human, rat and mouse P-selectin. Another three mAbs (P-sel.KO.2.2, P-sel.KO.2.11 and P-sel.KO.2.12) blocked the attachment of HL60 cells to P-selectin-transfected COS cells, demonstrating that these mAbs inhibit P-selectin-mediated adhesion. MAb cross-blocking experiments showed that these three mAbs bind to very close and overlapping epitopes. An ELISA assay using mAbs P-sel.KO.2.3 and P-sel.KO.2.12 was designed to measure soluble rat, mouse and human P-selectin. These anti-P-selectin mAbs are unique since they recognize common epitopes conserved during mammalian evolution and they may be useful for studying P-selectin function in inflammatory models in various species. PMID- 11019912 TI - Lectin purified human class I MHC-derived peptides: evidence for presentation of glycopeptides in vivo. AB - Previously, using synthetic glycopeptides carrying a natural cytosolic type of monosaccharide O-beta-linked N-acetylglucosamine (GlcNAc) glycosylation of serine residues, we have shown that glycopeptides act as suitable substrates for TAP mediated transport into the endoplasmic reticulum (ER), and that they bind efficiently to class I major histocompatibility complex (MHC) molecules and can elicit glycopeptide-specific cytotoxic T-lymphocyte (CTL) responses in mice. Recently, we have reported that peptides presented by human class I MHC molecules in vivo encompass a small but significant amount of peptides which seem to be carrying O-beta-linked monosaccharide GlcNAc. In the present report we provide further evidence that glycosylated peptides are indeed presented by class I MHC molecules in vivo. Thus, peptides derived from HLA-A*0201 were purified by wheat germ agglutinin (WGA) lectin affinity chromatography as previously described. Subsequently, the peptides contained in the WGA-eluate were subjected to sequence analysis by Edman degradation. It was found that the peptides derived from HLA A*0201 which had been retained by the O-GlcNAc-binding lectin WGA did indeed carry a HLA-A*0201 binding motif. Furthermore, using an enzymatic labeling procedure we present evidence that the HLA-A*0201-derived peptides which bind to the WGA lectin are glycosylated with terminal GlcNAc residues. Together, these data provide further evidence for the natural presentation by human class I MHC of glycopeptides carrying terminal O-GlcNAc residues in vivo. PMID- 11019913 TI - Polymorphism of the ABO blood group genes in Han, Kazak and Uygur populations in the Silk Route of northwestern China. AB - Genetic polymorphism in the ABO blood group gene of Han, Kazak and Uygur populations inhabiting the most northwestern part of China was investigated using polymerase chain reaction-based techniques. The present study enrolled 43 healthy unrelated Han, 37 Kazak and 59 Uygur volunteers. The allele in A1 blood group is distinguished A0101 and A0102 in difference of nucleotide position 467. The A0101 allele is more frequent in Caucasian and the A0102 allele is characteristic in Mongoloid. It must be notable that A0201 in the A2 group (with a single base deletion at nucleotides 1059 to 1061) which was characteristic of Caucasian was observed in Kazak and Uygur populations but not in Han. Further, 00201 (with no nucleotide deletion at 261 and three nucleotide differences), which is frequent in different races including Caucasian except for Mongoloid, was detected also in Kazak and Uygur populations. The frequencies of B0101 in Kazak, Uygur and Han were comparable to those of other Asian populations but higher than those of Caucasian populations. Collectively, these results reveal that the allele frequencies of Kazak and Uygur at the ABO blood group locus are an intermediate between those of Mongoloid and Caucasian, suggesting the admixed feature of Kazak and Uygur with Mongoloid and Caucasian. PMID- 11019914 TI - HNA-1a, HNA-1b, and HNA-1c (NA1, NA2, SH) frequencies in African and American Blacks and in Chinese. AB - The granulocyte antigens HNA-1a, -1b, and -1c (formerly named NA1, NA2 and SH) which reside on the neutrophil FcgammaReceptor IIIb (FcgammaRIIIb) play a major role in immune neutropenias and pulmonary transfusion reactions. In an attempt to shed some light on the origin and history of these antigens we typed the DNA of Blacks from South Africa (n=99), and Ghana (n=27), of 56 African Americans, and of 138 Chinese from Taiwan for HNA-1a,-1b, and -1c antigens using polymerase chain reaction with sequence-specific primers (PCR-SSP). In African and American Blacks, the HNA-1b antigen was more frequent than HNA-1a (77 vs. 67% and 77 vs. 59%, respectively). In contrast, in Chinese HNA-1a was more frequent than HNA-1b (91 vs. 54%). We observed 3 individuals with FcgammaRIIIB deficiency among the 126 tested African Blacks indicating a higher frequency of FcgammaRIIIB deficiency in Blacks than the reported 0.1% in Europeans. In addition, the frequency of HNA-1c in African and American Blacks (38 and 23%, respectively) was higher than the reported 5% in Europeans. Among the 57 HNA-1c (+) Blacks, all were HNA-1b (+) but only 26 were HNA-1a (+) supporting the idea that the HNA-1c antigen is the result of an additional point mutation in the allele coding for HNA-1b. Recently, HNA-1a, -1b, and -1c (+) Europeans have been reported to have three distinct FcgammaRIIIB genes. Among 26 Blacks who had been typed HNA-1a,b,c (+) by PCR-SSP we identified only 7 having three FcgammaRIIIB genes by DNA sequencing. When we sequenced the DNA of 6 HNA-1a,b,c (+) Europeans we found 4 of the individuals had three FcyRIIIB genes. Therefore, we assume that in Africa the point mutation occurred first in the HNA-1b allele resulting in the HNA-1c allele and the FcgammaRIIIB gene duplication took place later. PMID- 11019915 TI - Presence of a protective allele for achalasia on the central region of the major histocompatibility complex. AB - Idiopathic achalasia is a motility disorder of the esophagus whose etiology is unknown. An association between HLA genes and susceptibility to achalasia which suggests a possible immunogenetic mechanism has been reported recently. This study was designed to examine the HLA class II association in a large group of achalasia patients further and to investigate the distribution of TNFa and TNFb microsatellites in these patients. The study population, all Spanish, white and unrelated, consisted of 115 consecutive patients and 339 healthy controls. All of the patients had been diagnosed with primary achalasia of the esophagus with manometric, radiographic and endoscopic studies. All studies were performed on DNA samples after locus-specific amplification with the polymerase chain reaction: HLA-DRB1, DQA1 and DQB1 were typed by dot-blot hybridization and the size of the TNFa and TNFb microsatellites was measured using a semiautomatic method. The broad allele HLA-DQ1 was seen to be weakly associated with achalasia. The TNFa11 allele and the DRB1*1501-DQA1*0102-DQB1*0602 haplotype were reduced in achalasia patients but the stratified analyses showed that this was true only when both were present in the same individual. These results confirm the association between achalasia and HLA-DQ1 allele and suggest that TNFa11 is a marker for a protective allele for the disease, present on the B7-DRB1*1501 (7.1) ancestral haplotype in our population. PMID- 11019916 TI - Study of host- and virus-related factors associated with spontaneous hepatitis C virus clearance. AB - Epidemiologic parameters, virologic characteristics and frequency of HLA class II DR and DQ antigens were compared between 63 subjects with spontaneous hepatitis C virus clearance (group 1) and 282 patients with chronic active hepatitis C virus infection (group 2). DRB1*1101 and moreover DQB1*0301 alleles were more frequent in group 1 than in group 2 (33.8% vs. 14.7% and 64.4% vs. 28.6%; P=0.012 and P=0.003, respectively). The frequency of DQB1*02 was lower in group 1 than in group 2 (25.4% vs. 49%; P=0.04). No difference was observed in viral genotype distribution between group 1 and group 2. Univariate analysis showed that female sex and contamination by intravenous drug use were associated with self limited infection. However, by multivariate analysis, the only independent factor associated with hepatitis C virus RNA clearance was female sex (P=0.004). In conclusion, spontaneous hepatitis C virus RNA clearance is determined by class II antigens (mainly DQB1*0301) and female sex, while viral genotype plays no role. PMID- 11019917 TI - Identification of two novel human CD1E alleles. AB - CD1 is a family of proteins structurally related to major histocompatibility complex (MHC) molecules and specialized in presenting lipids or glycolipids to T cells. In humans, there are five CD1 genes (CD1A to CD1E). It has been shown that, in contrast with classical MHC genes, CD1 loci display a very limited polymorphism. In the present work we describe two novel CD1E alleles found in two healthy Caucasian individuals. One allele differs from the wild-type by a point mutation resulting in a replacement of arginine at position 154 by a tryptophan. In the second allele we found a substitution of the leucine 184 by a proline. PMID- 11019918 TI - Polymorphisms in the TNFA promoter region is not associated with palmoplantar pustulosis. AB - Polymorphisms of the 5'-flanking promoter/enhancer region of the TNAFA gene were determined in 80 Japanese patients with pulmoplantar pustulosis (PPP). The 5' flanking region of the TNFA gene from -1107 to 66 was amplified by polymerase chain reaction (PCR) method. Nucleotide sequencing data from the PCR products revealed that 5 single nucleotide polymorphisms at position 1031, -863, -857, 307 and -237. None of the nucleotide substitutions were significantly increased in PPP patients when compared with those in controls. To clarify the linkage among the neighboring genetic marker, we analyzed the association between the polymorphisms in the TNFA promoter region and the NcoI polymorphism in the first intron of the TNFB gene as well as HLA-DR9. The genotype at 1031C is strongly associated with TNFB1 and negatively associated with TNFB2 which is reported to be associated with PPP. These data indicate that TNFA gene centromeric to TNFB is not associated with PPP and the susceptible gene of PPP is located between TNFB and HLA-B. PMID- 11019920 TI - A study on the polymorphism of human MHC class I-related MR1 gene and identification of an MR1-like pseudogene. AB - Human MR1 is a recently discovered, ubiquitously transcribed gene very similar to the HLA class I loci and of unknown function. Mouse and rat MR1 sequences have also been described showing high similarity with the human gene. The goal of this work was to investigate if human MR1 was polymorphic. We have found that DNA sequences of MR1-specific polymerase chain reaction (PCR) products obtained from samples of diverse ethnic origin were invariant except in one case in which two silent mutations were detected. We also found an MR1-like sequence displaying significant differences with the previously described, the most remarkable of which is a STOP codon in the alpha2 domain indicating that is a pseudogene. PMID- 11019919 TI - Cloning and sequencing of canine MAGE cDNA. AB - Melanoma antigens (MAGE) are regarded to induce tumour-specific immune response and thought to be potential therapeutical agents for cancer immunotherapy. We hereby report the canine MAGE cDNA cloned from the testis of a beagle dog. Canine MAGE cDNA is 1,455 base pair (bp) nucleotides in length, and contains an open reading frame (ORF) of 1,137 bp nucleotides encoding a protein of 378 amino acids. The predicted amino acid sequence has 22-49% of homology with other MAGE proteins. mRNA transcripts of canine MAGE were detected only in the melanoma and testis and not in other normal tissues of adult dog by reverse transcriptase polymerase chain reaction (RT-PCR), indicating that the expression pattern of canine MAGE mRNA is similar to that of the MAGE family genes in tumor and normal tissues. PMID- 11019922 TI - Identification of a new HLA-B*39 allele: HLA-B*3924. AB - We have identified a new HLA-B*39 allele through polymerase chain reaction (PCR) using sequence-specific primers (SSP) and sequence-based typing of exons 2 and 3. This novel allele was identified in three HLA-identical siblings of Turkish origin. This allele only differs from HLA-B*3903 at a unique single nucleotide substitution (T for C) at position 365 in exon 3 which results in an amino acid change in codon 98 of methionine (ATG) to threonine (ACG). The sequencing enabled the development of a monospecific PCR-SSP reaction which can be used to discriminate between HLA-B*3924 and other B*39 alleles. PMID- 11019921 TI - HLA-B44 subtyping in the Catalan population using reference strand mediated conformation analysis. Implications for the selection of unrelated bone marrow donors. AB - Cytotoxic T lymphocytes (CTLs) reactive against the disparity between HLA-B*4402 and HLA-B*4403 have been reported after unrelated donor bone marrow transplantation. These CTLs have been associated with acute graft-versus-host disease and graft rejection. This study describes the HLA-B44-subtyping in the Catalan population using reference-strand mediated conformation analysis. It has been performed on 297 unrelated HLA-B44+ cord blood units from the Barcelona Cord Blood Bank (Barcelona, Spain). We have found a predominance of HLA-B*4403 (66.04%) over HLA-B*4402 (33.02%), whereas the predominant HLA-B44 allele in Northern Europe and the United States is HLA-B*4402. This inverted proportion between HLA-B44 subtypes in Mediterranean populations compared with other Caucasian populations suggests that HLA-B44 subtyping should be performed when an HLA-B44+ unrelated donor marrow is identified. PMID- 11019923 TI - HLA-B*4012: a new allele with unique serological features. AB - We have defined the new allele HLA-B*4012, which had been isolated from a black individual. It was initially recognized as a serologically unique allele when typing her father for renal transplantation. The HLA class I phenotype was A*0201,*6602; B*4001,*4012; Bw6; Cw*0304,*1505. Sequencing from exon 1 through intron 3 of B*4012 was performed. B*4012 is identical to B*4001 and B*4010 in exon 3, and in the 3' part of exon 2, but it is unique in that exon 1 and the 5' part of exon 2 are identical to B*1503, B*1509, B*1510, B*1518, B*1523, and B*1529. The generation of this allele is best explained by a recombination event in exon 2 (break point between nucleotides 205 and 222 from the beginning of the coding region) of B*4001 or B*4010 with one of these B*15 variants as a donor allele. Its unique serological feature (B48, B60, B70, and B72 reactivity) is consistent with the sequence data of its donor alleles. PMID- 11019924 TI - Identification of a new HLA-B*78 allele in a Hispanic family. AB - A new B*78 variant, B*7804, was detected in three members of a Hispanic family. The novel B*78 sequence differs from B*78021 by two substitutions: T at nucleotide 527 (all other B*78s have A) and T at nucleotide 583 (all other B*78s have a C). Both nucleotide substitutions encode amino acid changes at codons 152 and 171, respectively. PMID- 11019925 TI - HLA-B*8202 identified in a Caucasoid potential bone marrow donor. AB - Sequence analysis of HLA class I alleles has continued to reveal the true extent of polymorphism, particularly for B-locus alleles. This diversity can arise through reshuffling of polymorphic sequences generated by point mutation, resulting in interallelic recombination or intergenic recombination (1). Here we describe a new B-locus allele, B*8202, which is structurally most similar to B*8201, having only one nucleotide difference in exon 3 at nucleotide 557, resulting in an amino acid change of aspartic acid to glycine at residue 162. Glycine is the consensus amino acid for B-locus alleles, which suggests that B*8202 is older than B*8201 in evolutionary terms. B*8201 was found to be a hybrid of B*4501 and B*5602 that may have arisen through recombination events, explaining the serological patterns observed with these allotypes. The importance of high-resolution typing is emphasised here as routine typing suggested the presence of B*8201 and the new variant allele may have been missed had it not been typed further by sequence-based typing. PMID- 11019926 TI - New HLA-DRB1* nucleotide sequence of Italian origin: HLA-DRB1*13022. AB - High-resolution polymerase chain reaction using sequence-specific primer typing of the HLA-DRB1 gene of an Italian patient waiting for unrelated bone marrow transplantation revealed a new allelic variant of HLA-DRB1*13. Sequencing the exon 2 of DRB1* gene demonstrated a G-->C transition at the nucleotide 216 resulting in a silent mutation at codon 72: CGG-->CGC. The closest sequence was the HLA-DRB1*1302 and the new allele was named HLA-DRB1*13022. This variant was carried by the haplotype HLA-A*24; Cw*0702; B*39; DRB1*13022; DRB3*0301; DQA1*0102; DQB1*0604 as demonstrated by a family study. PMID- 11019927 TI - Identification of a novel allele, DRB1*1340, in two Brazilian individuals. AB - We identified a novel HLA-DRB1 allele, named DRB1*1340 by the WHO HLA Nomenclature Committee, in two Brazilian individuals. Typing by polymerase chain reaction using sequence-specific oligonucleotide probes (PCR-SSOP) showed a DRB1*13 allele with an unusual hybridization pattern. DNA sequencing of both strands and comparison of the sequence with previously described DRB1 alleles revealed that the most similar allele is DRB1*1301, from which DRB1*1340 differs by a single nucleotide (T-->A) in exon 2, at position 127, codon 47 (Phe-->Tyr). The sequence received accession number AJ237964 from the EMBL database. PMID- 11019928 TI - A new HLA-DPA1 allele, DPA1*02016, identified in African-American population. AB - A new HLA-DPA1 allele has been found in the African-American population. This newly discovered type is homologous with the DPA1*02011 allele, except in codon 38 of the DPA1 sequence. The variant of DPA1 was detected by sequence-specific oligonucleotide (SSO) typing, and specified by cloning and sequencing methods. GenBank accession number is AF165160. The WHO Nomenclature Committee has officially assigned the name DPA1*02016. PMID- 11019929 TI - Antidepressant selection: proceedings from a TCA/SSRI Concensus Conference. AB - OBJECTIVE: Despite significant advances in treatment choice, the use of antidepressants in clinical practice remains far from optimal. One aspect of antidepressant use, initial antidepressant selection, is the most immediately influential way to improve clinical outcomes. The purpose of this conference was to review the available literature on the two most common classes of antidepressants--the tricylic antidepressants (TCAs) and the selective serotonin reuptake inhibitors (SSRIs)--and come to consensus on recommendations for initial antidepressant selection. METHOD: A panel of experts in psychiatry and health services research was convened to systematically review and discuss the available literature on the TCAs and SSRIs in the treatment of depression and related affective disorders. RESULTS: The consensus group concluded that growing evidence suggests that TCAs are no longer justified as first-line antidepressant therapy in most settings. In addition, emerging differences in the tolerability profiles and breadth of indication of the SSRIs are important to consider when making an initial treatment choice. CONCLUSION: While the coming years will bring new treatment options, clinicians currently have the ability to improve clinical outcomes and the long-term prognosis of depression. The following pages provide information to make informed antidepressant choices, thereby providing a higher quality of antidepressant treatment for the patient in actual clinical practice. PMID- 11019930 TI - Treatment of depression with associated anxiety: comparisons of tricyclic antidepressants and selective serotonin reuptake inhibitors. AB - OBJECTIVE: Evidence indicates that alleviating anxiety symptoms early in the treatment of depression improves treatment compliance, limits treatment discontinuations and contributes to a positive treatment outcome. Because of their sedating effects, the tricyclic antidepressants (TCAs) have historically been the first-choice agent in treating comorbid depression and anxiety. However, a growing body of evidence suggests that the selective serotonin reuptake inhibitors (SSRIs) can also be suitable therapy. METHOD: We reviewed literature regarding the use of TCAs and SSRIs in depressed patients with comorbid anxiety. RESULTS: SSRIs are at least as effective as TCAs in the treatment of both overall depression as well as anxiety symptoms. TCAs can cause significant and sometimes unacceptable side effects which limit their therapeutic potential. SSRIs, on the other hand, have little or no effect on cholinergic, histaminergic or adrenergic receptors, and have a very favourable tolerability profile. CONCLUSION: The traditional selection of antidepressants based on the presence or absence of anxiety has little scientific support. In considering the overall risk:benefit ratio, SSRIs should be the first line treatment for depression with associated anxiety. PMID- 11019931 TI - Safety and tolerability considerations: tricyclic antidepressants vs. selective serotonin reuptake inhibitors. AB - OBJECTIVE: An important consideration in the choice of an antidepressant is its safety and tolerability. METHOD: We present a review of literature, clinical trials and meta-analyses regarding the safety and tolerability of the tricyclic antidepressants (TCAs) and the selective serotonin reuptake inhibitors (SSRIs) in depressed patients. RESULTS: The SSRIs have a very favourable side-effect profile compared to the TCAs and are associated with fewer treatment discontinuations. Unlike the TCAs, they do not cause anticholinergic, hypotensive or sedating reactions, and are not associated with impaired cognitive function. Their most common side-effects (nausea, vomiting, nervousness, insomnia, headache and sexual dysfunction) are usually mild and typically disappear as treatment continues. The SSRIs also exhibit lower toxicity and lower lethality when taken in an overdose situation. Although the safety profiles of the principal SSRIs appear to be comparable, there is some data showing important differences in the severity and frequency of specific adverse events. CONCLUSION: The SSRIs have a more favourable safety profile than the TCAs in both acute and long-term treatment of major depression. PMID- 11019932 TI - Tricyclic antidepressants and selective serotonin reuptake inhibitors: use during pregnancy, in children/adolescents and in the elderly. AB - OBJECTIVE: Depressive disorders can occur at any point in the lifespan. One way of differentiating antidepressants is by examining their efficacy and safety in the special patient populations that exist along the lifespan. Thus, we examine clinical data that is available regarding the use of selective serotonin reuptake inhibitors (SSRIs) and tricyclic antidepressants (TCAs) at three distinct points in the lifespan: childhood and adolescence, pregnancy and late adulthood. METHOD: Literature regarding the use of antidepressants in children and adolescents, pregnancy and the elderly was reviewed. RESULTS: Clinical data suggest that SSRIs should be first line treatment in children and adolescents as TCAs have questionable efficacy and definite safety issues. Similarly, although TCAs and SSRIs show equivalent efficacy in elderly patients, the safety profile of the SSRIs makes them a more prudent choice in this population. Finally, although there is no definitive data that contraindicates the use of a particular antidepressant in pregnancy, the bulk of reassuring pregnancy outcome data exists for the SSRIs, specifically for fluoxetine. CONCLUSION: Although no single antidepressant can ever be recommended for every patient, SSRIs should be considered the first-line choice in the treatment of depression in special patient populations. PMID- 11019933 TI - Antidepressants in long-term therapy: a review of tricyclic antidepressants and selective serotonin reuptake inhibitors. AB - OBJECTIVE: Although depression has been shown to be a long-term disorder, most research studies have concentrated on its acute treatment. METHOD: A literature review of the use of tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs) in long-term treatment of depression was performed and recommendations regarding long-term treatment were summarized. RESULTS: Studies conclusively document the need for continuation treatment after initial remission of symptoms to prevent relapse. Studies also suggest that continuation treatment should last a minimum of 3-6 months following acute response. Conclusions from a few maintenance studies clearly show that recurrence rates are lower when patients at risk for recurrence continue their active treatment at its original dose than when they are switched to placebo. CONCLUSION: Overall, studies conclude that depression is a recurrent, often chronic, lifetime illness requiring long-term treatment. Continuation therapy of 3 to 6 months after acute stabilization should be considered standard for all depressed patients, and maintenance therapy should be considered for many depressed patients. Newer agents, such as the SSRIs, are preferable to the TCAs for long-term treatment based on their superior tolerability and safety. However, because of the limited number of maintenance studies, further studies using a prospective, randomized design are needed to address this issue. PMID- 11019934 TI - Anxiety disorders: a review of tricyclic antidepressants and selective serotonin reuptake inhibitors. AB - OBJECTIVE: Anxiety disorders are the most common mental health disorders. While the older tricylic antidepressants (TCAs) are efficacious in the treatment of many anxiety disorders, recent studies with fluoxetine and other selective serotonin reuptake inhibitors (SSRIs) have emphasized the role of serotonin in the aetiology of these conditions. METHOD: We reviewed the efficacy, safety and tolerability of TCAs and SSRIs in the treatment of the most common anxiety disorders, specifically, panic disorder, obsessive-compulsive disorder, post traumatic stress disorder, social phobia and generalized anxiety disorder. RESULTS: Both the TCA and SSRI antidepressants are effective in treating a wide variety of anxiety disorders. SSRIs, due to their greater safety and tolerability, should be the preferred choices in treating anxiety disorders in those instances where TCAs and SSRIs are considered equally effective. In the cases of OCD and social phobia, SSRIs are almost always preferable given that the TCAs do not appear effective in these disorders. CONCLUSION: Further research is needed on the naturalistic long-term use of the TCAs and SSRIs in the treatment of anxiety disorders. PMID- 11019935 TI - Compliance with antidepressant therapy and antidepressant discontinuation symptoms. AB - OBJECTIVE: Compliance with antidepressant medication is important in order to achieve all the goals of antidepressant therapy. These goals include symptom resolution, restoration of normal functioning and prevention of relapse or recurrent episodes. METHOD: We discuss compliance and review adverse symptoms that may occur when antidepressant treatment is abruptly discontinued or interrupted. RESULTS: The physician's role in managing compliance includes preventing or minimizing the risk for discontinuation symptoms, counselling patients regarding the risk for symptoms if doses are missed and choosing pharmacotherapy that is forgiving of non-compliance. Physicians also need to recognize the symptoms of discontinuation syndrome in order to differentiate patients who are non-compliant from patients who require a dosage adjustment or adjunctive therapy. Finally, physicians need to understand how to treat discontinuation symptoms when they do occur. CONCLUSION: Since published data indicate that between 30% and 60% of patients do not take their medications as prescribed, non-adherence to antidepressant medication is likely to be a significant clinical issue in the management of many patients. PMID- 11019936 TI - Antidepressant use patterns in clinical practices: comparisons among tricyclic antidepressants and selective serotonin reuptake inhibitors. AB - OBJECTIVE: Antidepressant use in clinical practice may differ from that in randomized controlled trials due to the complex interaction between patients, prescribers and the health-care system. METHOD: A review of studies using data from actual clinical practice has found consistent differences in the prescribing patterns of tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs). RESULTS: Patients who initiate therapy on SSRIs are more likely to complete a course of adequate dose and duration of antidepressant therapy than patients who initiate therapy on a TCA. Differences in prescribing patterns have also emerged among the most common SSRIs, fluoxetine, sertraline and paroxetine, that are also consistent over time, settings and countries. CONCLUSION: Given that the drugs studied are one constant across all the studies, the antidepressants' underlying pharmacological differences in tolerability and other properties may provide an explanation for differences in prescribing patterns. PMID- 11019937 TI - Economic outcomes associated with tricyclic antidepressant and selective serotonin reuptake inhibitor treatments for depression. AB - OBJECTIVE: To compare the economic outcomes associated with the tricyclic antidepressants (TCAs) and the selective serotonin reuptake inhibitors (SSRIs) in the treatment of depression. METHOD: A literature review of pertinent studies was performed. The advantages and disadvantages of clinical trials versus observational studies are described, and the breadth of the economic outcome measure chosen for the conclusions reached is discussed. RESULTS: The inclusion and exclusion criteria of clinical trials, in combination with their strict provider and patient study protocols, limit their generalizability to naturalistic treatment settings. Retrospective studies of patients can provide valuable information about the experiences and costs incurred by patients in actual treatment. However, confounding factors (both observable and unobservable) limit the amount of confidence that can be placed in inferences about treatment effects. Randomized prospective studies with naturalistic follow-on may help to mitigate some of the concern about treatment confounders which has traditionally been associated with non-randomized observational studies. CONCLUSION: Retrospective studies and one randomized prospective study of the economic outcomes of TCA versus SSRI treatment have found the SSRIs to be less expensive than TCAs when total direct medical expenditures are considered. However, additional studies are needed to address this issue. PMID- 11019938 TI - A full-UV spectrum absorbing daily use cream protects human skin against biological changes occurring in photoaging. AB - BACKGROUND: There is overwhelming evidence that exposure of human skin to ultraviolet radiations (UVR) leads to the development of cutaneous photoaging and eventually to neoplasia. This study was designed to evaluate in humans the protection afforded by a daily use cream containing a photostable combination of UVB and UVA absorbers (Uvinul N539, Parsol 1789 and Mexoryl SX) providing a continuous absorption through the entire UV spectrum, against damages induced by repeated daily exposure to solar simulated radiation (SSR). METHODS: Buttock skin of 12 healthy volunteers was exposed 5 days per week for 6 weeks to one minimal erythema dose of solar simulated radiation per exposure. The following parameters in treated and untreated skin were evaluated: erythema, pigmentation, skin hydration, skin microtopography, histology and immunochemistry, and collagen and metalloproteinase (MMP) mRNA levels. RESULTS: In SSR exposed unprotected skin sites, we observed melanization and changes in the skin hydration and microtopography. The epidermis revealed a significant increase in stratum corneum and stratum granulosum thickness. In the dermis, an enhanced expression of tenascin and a reduced expression of type I pro-collagen were evidenced just below the dermal epidermal junction. Although we were unable to visualize any change in elastic fibers in exposed buttock skin, a slightly increased deposition of lysozyme and alpha 1 antitrypsin on these fibers was observed using immunofluorescence techniques. Furthermore, types I and III collagen mRNA were slightly increased and a significant enhancement (up to 2.8-fold) of MMP-2 mRNA level was observed. The daily use cream was shown to prevent all these biological changes. CONCLUSION: Our results show in vivo that an appropriate full-UV spectrum product significantly reduces the solar-UV-induced skin damage, demonstrating the benefit of daily photoprotection. PMID- 11019939 TI - Interaction of commonly used emollients with photochemotherapy. AB - BACKGROUND/AIM: Even though emollients are commonly used in combination with photochemotherapy, we still lack guidance for the selection of the appropriate emollient to be used in combination with photochemotherapy. The purpose of our study was to determine the interaction of commonly used emollients with photochemotherapy. METHODS: The study was carried on 75 healthy volunteers. In order to assess the effects of five different emollients--white petrolatum, 3% salicylic acid in white petrolatum, Balmandol, Decubal and Urederm hydro--the subjects were divided into five groups. Minimal phototoxic dose (MPD) was determined by irradiating two rows of six circular test fields on the back of the subjects with increasing doses of UVA. One of the emollients listed above was applied to the skin under the upper rows alternatively, whereas the lower rows served as control. RESULTS: Application of Urederm hydro and Decubal before exposure to UVA resulted in a statistically significant decrease in the MPD in all subjects. Even though application of white petrolatum and white petrolatum containing 3% salicylic acid decreased the MPD in most of the subjects and Balmandol application slightly increased the MPD, the results did not reach statistical significance. CONCLUSIONS: According to our findings, it is probable that the total dose of UVA needed to cause clearing of psoriasis would be decreased by application of Urederm hydro and Decubal prior to photochemotherapy. We noted a slight photoprotective effect of Balmandol in the UVA range, but this finding has to be further studied. Clinical studies are needed to clarify the effects of these emollients on the course and outcome of photochemotherapy. PMID- 11019940 TI - Polychromatic phototest as a prognostic tool for polymorphic light eruption. AB - BACKGROUND: Diagnosis of polymorphic light eruption (PLE) is based on the patient's history, the morphology of the lesions and the results of phototesting. Skin lesions of PLE can be provoked by repetitive UVB or UVA irradiation. However, about 20% of the patients with PLE have negative phototests. As 24% of the patients with PLE go into remission, it was of interest to search for a link between the results of the phototests and the evolution of the photodermatosis. METHODS: Forty patients with PLE were recruited and repetitive phototests were performed. To ensure a good reproducibility of the phototests, one to three phototests were performed on each patient at different stages of the disease including the period when the PLE had gone into remission. RESULTS: Except for one patient, there was a good reproducibility of the repetitive polychromatic phototests: in each patient, the tests remained positive or negative throughout the disease. After long-term follow-up, two different subgroups were identified: 30 patients with active PLE and 10 patients in remission. There were no clinical differences between these two groups apart from the age of onset and the clinical lesions of the PLE. PLE began at an earlier age in the patients in remission and presented mainly with a plaque-type eruption. In total, 52.5% of the patients had at least one positive polychromatic phototest. Phototests were positive only in patients with active disease. All the patients in remission had negative phototests. CONCLUSIONS: Repetitive phototests could be a prognostic marker for PLE. Two subtypes of PLE were identified on the basis of phototest results: the benign form of PLE with negative phototests, which tends to go into remission, and the more severe and more chronic PLE, with positive phototests. PMID- 11019941 TI - Comparative quantitative analysis of ultraviolet B-induced skin blood flow change using laser Doppler perfusion imaging technique. AB - Laser Doppler perfusion imaging (LDI), recently developed, can generate a color coded image of tissue perfusion, making it possible to assess the spatial distribution of skin perfusion without touching the surface. Using this apparatus, we investigated ultraviolet B (UVB)-induced perfusion profile of the skin and compared the results with those obtained from laser Doppler flowmetry (LDF). Fifteen adult male Korean volunteers were irradiated with several doses of UVB ranging from 25 mJ/cm2 to 186 mJ/cm2. Twenty-four hours later, the erythema reaction was evaluated with LDF and LDI systems. There was a significant correlation between the logarithmic dose of UVB and erythema values. The curves consisted of two parts, an initial, flat phase and then a linear, steep one. Also, there was a good correlation between LDF and LDI. The LDI is as sensitive as conventional LDF, but has the many advantages of measuring blood flow over large areas without contact with the skin surface. This instrument will be useful in the measurement of skin blood flow in many areas of dermatological application. PMID- 11019943 TI - Effects of a 2-week treatment with pulsed monochromatic light in healthy pigs: a clinical and morphological study. AB - BACKGROUND: Pulsed monochromatic light (PML) is now used clinically for pain relief and wound healing in both human and veterinary practice. The purpose of this study was to evaluated the clinical and pathological effects of PML irradiation in an animal model, using healthy specific pathogen-free pigs. METHODS: After 2 weeks of habituation, one group of animals (n=9) underwent treatment with pulsating monochromatic infrared and red light while the control group (n=9) was left untreated. PML was given five times a week during a 2-week period, and at each treatment the total radiant exposure was 6.3 J/cm2. At the completion of the study, all pigs were subjected to complete necropsy. RESULTS: None of the animals showed any clinical signs of disease during the study period. The measured hematological and clinico-chemical variables all showed values within the reference range and the daily weight gain was high in both the treatment and control groups (825 and 923 g/day, respectively). The pathological examination revealed no morphological differences between treated and non-treated animals. CONCLUSION: In healthy pigs, no adverse effects of low-energy photon therapy on the clinical state of health or on the morphology of different tissues were observed. PMID- 11019942 TI - p53, cyclin-D1, PCNA, AgNOR expression in squamous cell cancer of the lip: a multicenter study. AB - BACKGROUND: The development of squamous cell carcinoma of the lower lip is an interesting model of photocarcinogenesis because of the structural and topographic characteristics of the lips. The purpose of this study was to evaluate the expression of immunohistochemical markers on the lips of patients with lower lip squamous cell carcinoma (LLSCC), compared with a control population. METHODS: Of the 98 subjects involved in the study, 58 were suffering from squamous cell carcinoma of the lower lip. The remaining 40 acted as a control. The case studies were taken from six university and hospital dermatology and plastic surgery departments. Questionnaires were administered to assess the risk factors for LLSCC. The cases involving squamous cell carcinoma underwent surgical excision and punch biopsy specimens were obtained from 20 control patients. Tissues were prepared in 5-microm-thick sections to carry out the following immunohistochemical study: PCNA, p53, AgNOR, cyclin-D1, bcl-2. RESULTS: The lower lip was the predominant location of squamous cell carcinoma, with the following factors playing important roles: chronic sun exposure, history of smoking, alcohol use and familial risk of cutaneous tumors. The male/female ratio in our survey was 5:1. The p53 protein was positive in approximately 50% of SCC cases and in 20% of controls. This protein is mostly associated with chronically photoexposed skin areas. AgNOR positivity increased with the loss of cellular differentiation; a progressive increase in size and a poorly defined shape were evident in poorly differentiated carcinomas. CONCLUSIONS: The results of this multicenter study showed that there is a noticeable difference in the expression of PCNA, p53, cyclin-D1, and AgNOR in tissues from patients with LLSCC and controls. PMID- 11019944 TI - Correlation between bathing time and photosensitivity in 8-methoxypsoralen (8 MOP) bath PUVA. AB - Bath PUVA (psoralen plus ultraviolet A) using 8-methoxypsoralen has become increasingly popular in recent years as an effective treatment option for a continuously expanding range of skin disorders. Among the various variables of bath PUVA treatment, the impact of bathing time on photosensitivity has never been investigated in detail. We therefore determined the threshold UVA dose for erythema induction after different bathing periods. A marked influence of bathing time on photosensitivity was found. Increasing the soaking period from 5 min to 30 min resulted in a greater than 60% reduction of the minimal phototoxic and minimal perceptible phototoxic dose. Our results demonstrate that the duration of the psoralen bath is a critical parameter in bath PUVA treatment and has a major influence on UVA dose requirements. PMID- 11019945 TI - Reduction in 8-methoxypsoralen immersion time alters the erythemal response to bath PUVA. AB - Topical psoralen plus ultraviolet A (PUVA) using 8-methoxypsoralen (8-MOP) bath solution is a well established and effective treatment in dermatology. The standard immersion time in the UK is 15 min, but a shorter bathing period could potentially increase treatment convenience. In order to examine the effect of reduction in immersion time on skin phototoxicity, we compared the erythemal response to UVA following 5 min and 15 min psoralen baths. The study was performed on the forearm skin of 7 healthy volunteers using an 8-MOP psoralen concentration of 2.6 mg/l. One forearm of each volunteer was soaked for 15 min and the other for 5 min, followed by immediate irradiation with a series of 10 doses of broadband UVA ranging from 0.1 J/cm2 to 6.9 J/cm2. At 72 h, the minimal phototoxic doses (MPDs) were noted and erythema readings (erythema index) were taken in triplicate with a reflectance instrument. The median MPD following 5 min immersion was 1.7 (range 0.7-2.7) J/cm2 compared with 1.0 (range 0.4-1.7) J/cm2 after 15 min treatment, with no significant difference. However, the mean slope of erythema dose-response on the 15-min treated side was significantly steeper than on the 5-min treated side, 0.036 and 0.021 respectively, P < 0.05. Hence, this preliminary work shows that reducing 8-MOP immersion time to 5 min reduces the erythemal response to UVA. It will clearly be necessary to examine the effect of a shortened immersion period on disease clearance before considering such a change to the topical PUVA regime. PMID- 11019946 TI - Transmission of UVA radiation through epithelium of oral mucosa and skin in rat and man. AB - BACKGROUND/METHODS: Transmission of 362 nm ultraviolet A radiation (UVA) through epithelium of oral mucosa and skin epidermis was compared in rat and man. The transmitted irradiance was measured by means of a radiometer. RESULTS: In man, significantly (P<0.01) greater transmittance (2.8-fold) was observed in skin compared to buccal mucosa. In rat, no significant differences were found. CONCLUSION: In different species and different types of epitheliae, separate studies are needed for the evaluation of UVA transmission. PMID- 11019947 TI - What's new in photodermatology. PMID- 11019948 TI - Quinacrine sterilization: experience among women at high risk for surgery. AB - A non-surgical method of female sterilization is needed because many women are at high risk of complications with standard surgical methods, especially in developing countries. Also, some women who desire sterilization refuse it for fear of surgery. To meet these special needs, we initiated a trial of quinacrine sterilization (QS), a non-surgical method involving transcervical insertion of 252 mg quinacrine as pellets by a modified IUD inserter. Diclofenac (50 mg) was inserted with the quinacrine pellets. This insertion was repeated a month later and a 150-mg injection of depo medroxyprogesterone was administered at the time of the first insertion. One hundred and thirty-four women of reproductive age entered the trial. Of these, 92 were considered to be at high risk for surgery, 27 had refused surgery, and 15 had had failed surgical sterilization. Mean follow up was 3.46 years. No pregnancies or serious complications were experienced. The main side-effect was menstrual irregularity, due probably to the depo medroxyprogesterone injection. QS is a suitable option for women at high risk of surgical complications. PMID- 11019949 TI - Characteristics of copper corrosion in simulated uterine fluid in the presence of protein. AB - In order to acquire more knowledge of the performance of copper-bearing intrauterine devices (Cu-IUDs), corrosion behavior of copper in a simulated uterine fluid was investigated in the presence of proteins. The proteins studied included serum albumin, gamma-globulin and hemoglobin. Electrochemical polarization resistance measurements indicated that, in all cases under study, the corrosion rate of copper declined with time and the proteins always increased the rate. Moreover, different kinds of protein showed different dependence of copper corrosion rate on protein level. Addition of serum albumin initially raised the copper corrosion rate; however, the higher albumin concentration resulted in less promotion of the corrosion. X-ray diffraction (XRD) of the specimen surface showed that, both in the presence and absence of serum albumin, cuprous oxide was the only corrosion product, even though at higher concentrations of albumin less cuprous oxide was formed. This implies that serum albumin does not alter the corrosion mechanism. Electrochemical cathodic reduction of the oxide film and chemical analysis of solution after corrosion testing proved that the proportion of soluble ionic copper in the whole corrosion products varied with the albumin level. The copper corrosion rate in the presence of gamma-globulin or hemoglobin increased monotonically with increasing concentration of the protein. These three proteins shifted the corrosion potential of copper towards negative. It could be inferred that the proteins accelerated the anodic dissolution process of copper and hence enhanced its corrosion. PMID- 11019950 TI - Comparative study on the acceptability of two modern monophasic oral contraceptive preparations: 30 microgram ethinyl estradiol combined with 150 microgram desogestrel or 75 microgram gestodene. AB - Cycle control and tolerability of two monophasic oral contraceptive pills containing 30 microg ethinyl estradiol (EE) with either 150 microg desogestrel (DSG) or 75 microg gestodene (GSD) were compared in women starting oral contraception. A minimum of 200 healthy women at risk for pregnancy were to be treated for a total of 6 cycles per patient in a prospective, randomized open parallel-group multicenter trial. Two hundred and forty-one subjects were randomized, 115 to DSG/EE and 126 to GSD/EE. Compliance to the study preparation was high (around 95%) in both groups and no pregnancies occurred during the study. Cycle control was excellent; there were no differences between the two groups with regard to incidence of spotting and breakthrough bleeding or duration and intensity of withdrawal bleeding. Side-effects were mild and in general comparable in the two groups. Both at baseline and during treatment, a higher proportion of women taking GSD/EE complained about breast tenderness. This resulted in more early withdrawals because of breast tenderness in the GSD/EE group. It was concluded that monophasic DSG/EE and GSD/EE are equally effective, have similar cycle control and both are generally well tolerated. PMID- 11019951 TI - Performance of the frameless IUD (Flexigard prototype inserter) and the TCu380A after six years as part of a WHO multicenter randomized comparative clinical trial in parous women. AB - The clinical performance of the new Frameless IUD was compared with the TCu380A, the most widely used copper IUD in the world today. Insertions of the Frameless IUD were conducted with a prototype inserter (Flexigard). We report on the 6-year results from a randomized comparative clinical trial conducted at the Shanghai Institute of Planned Parenthood Research, as part of an international multicenter WHO clinical trial, involving 200 women, respectively recruited for use of each device. The cumulative 6-year pregnancy rates were 0.0 per 100 women for the Frameless IUD and 3.3 for TCu380A. Termination due to partial expulsion was significantly less for the Frameless IUD as compared with TCu380A (0.0 and 4.3 per hundred women, respectively). Complete expulsion, bleeding, pain, bleeding and pain and other medical reasons for termination did not differ significantly between the two devices. The net cumulative continuation rates at six years per 100 women were 80.8 for TCu380A, and 83.0 for the Frameless IUD. Both Frameless IUD and TCu380A are highly effective, safe and acceptable contraceptive devices, because of low pregnancy rates (per 100 women), and low termination rates due to expulsion, pain, bleeding, and bleeding and pain. Due to its anchoring, the Frameless IUD is significantly more effective than the TCu380A IUD as regards proper retention of the IUD in the uterine cavity. This may also explain its lower failure rate. PMID- 11019952 TI - Pilot study on the use of repeated doses of misoprostol in termination of pregnancy at less than 9 weeks of gestation. AB - Pregnancy was terminated by repeated doses of vaginal misoprostol in 20 women at a gestational age of less than 9 weeks. The women were given 800 microg of vaginal misoprostol as an initial dose followed by 400 microg of vaginal misoprostol every 3 h for 4 doses. Fourteen women (70%, 95% confidence interval: 48 85%) had a complete abortion. Two women (10%) had a missed abortion, and two (10%) had an ongoing pregnancy. Two women ( 10%) had an incomplete abortion. The interval between the first dose of misoprostol and the passage of tissue mass was 25.3 +/- 34.4 h (median: 15 h). The duration of vaginal bleeding was 23.6 +/- 20.4 days (median: 14 days). Side-effects were mild and there was no significant drop in hemoglobin level. Repeated doses of vaginal misoprostol may be an alternative for women who do not want surgical abortion and who live in an area where mifepristone is not available. PMID- 11019953 TI - Contraceptive knowledge, attitude and practices of men in rural Maharashtra. AB - Since men are the dominant decision makers in India, it is prudent to discover the knowledge, perception, attitudes and contraceptive practices of men to improve their involvement in the reproductive health needs of families. Three thousand and seventy-two married men from a tribal Primary Health Centre (PHC) area in Thane district of Maharashtra State, India were surveyed with special emphasis on investigating the reasons for not accepting male methods. The majority of them not only had no concept of family spacing, but had not even taken any initiative to improve their knowledge or acceptance of condom/ vasectomy. Men who were aware of contraceptive methods had little knowledge of their correct use. Of the men, 53.7% had positive views about their role in family planning while 66.2% of men stressed the need to improve the acceptance of male methods by providing knowledge and information through sources such as radio, television, door-to-door campaigning and interpersonal communications. Thirty per cent emphasized the need to improve the availability and quality of services. This study indicates a pressing need for effective intervention strategies, both at the community and the clinic level, backed with efficient counselling, motivation and provision of services in rural and remote areas. PMID- 11019954 TI - Contraceptive status and sexual behavior in women over 35 years of age in India. AB - The aim of this paper is to survey the current contraception use and analyze the sexual patterns in women above 35 years of age in India. Five hundred women, of whom 250 were at least 35 years old and 250 were less than 35 years old (control group) were interviewed with the help of a prepared questionnaire. A permanent surgical method of contraception had been accepted by 40.4% of women > or = 35 years old versus 16.8% of women < 35 years old. Of various temporary methods, an intrauterine device (IUD) was used by 5.2% and 22.8% oral contraceptive pills (OCs) by 1.2% and 9.6%, and natural methods by 6.4% and 3.2%, in women > or = 35 years and < 35 years, respectively. No contraception was used by 25.6% women > or = 35 years. A history of medical termination was given by 67.18% of non contraceptive users vs. 27.41% of contraceptive users in women > or = 35 years. Coital frequency was noted at 4.32 times/month in women > or = 35 years but 7.2 times/month in women <35 years. Among contraceptive users > or = 35 years, coital frequency was 4.9 times/month compared with 3.9 times/month in non-contraceptive users. There is a high acceptance of irreversible sterilization and a high abortion rate in women > or = 35 years. Use of contraceptives was linked with increased sexual activity and a decreased abortion rate. PMID- 11019955 TI - Control of oral anticoagulant therapy with whole blood prothrombin time devices: the future has begun. PMID- 11019956 TI - Plasma procarboxypeptidase U in men with symptomatic coronary artery disease. AB - Procarboxypeptidase U (proCPU) is the plasma precursor of carboxypeptidase U (CPU, carboxypeptidase R. plasma carboxypeptidase B or activated thrombin activatable fibrinolysis inhibitor, TAFIa). CPU removes C-terminal lysine residues that act as plasminogen binding sites from partially degraded fibrin, thereby down-regulating plasminogen activation and fibrinolysis. The present study was carried out as a pilot study to examine whether the plasma proCPU concentration is related to the presence of coronary artery disease (CAD) and/or to levels of established risk indicators for CAD, in a case-control study of 110 men requiring coronary artery bypass grafting (CABG) because of stable angina pectoris. The preoperative plasma proCPU level in the CABG patients was significantly higher than in population-based controls (1029 +/- 154 vs. 974 +/- 140 U/L, p <0.05). In addition, in a subset of the patients (n = 31 ) the proCPU concentration, which was significantly lower on the third postoperative day (-17 +/- 10%), had increased significantly on the sixth day (+14 +/- 12%) after surgery, compared with the preoperative level. In both patients and controls, proCPU concentration was strongly and positively associated with factor VII amidolytic activity and protein C activity, suggesting a common mechanism modulating the plasma levels of these proteins. Otherwise, statistically significant correlations with proCPU were group-specific. In the patients, proCPU correlated significantly with plasma fibrinogen and protein S. In the controls, proCPU correlated significantly with concentrations of cholesterol in plasma. VLDL and LDL. In addition, proCPU correlated significantly with C-reactive protein and haptoglobin levels in the controls only, indicating that also inflammatory mechanisms are involved in the regulation of plasma proCPU. These results suggest that a mechanism exists by which fibrinolytic function is impaired in a manner that is likely to result in more stable fibrin deposits and increase the risk of precocious CAD as well as early occlusion of venous bypass grafts. PMID- 11019957 TI - A new candidate missense mutation (Leu 1657 IIe) in an apparently asymptomatic type 2A (phenotype IIA) von Willebrand disease family. AB - Type 2A von Willebrand disease (VWD) is mostly an autosomal dominantly inherited bleeding disorder characterised by a qualitative defect of von Willebrand factor (VWF). Mutation screening was used to screen the whole of VWF gene followed by direct sequencing to detect the mutation in a father and son diagnosed with type 2A (phenotype IIA) von Willebrand disease. A C5219 to A transversion was detected predicting Leucine to Isoleucine substitution in codon 1657. This novel missense mutation which was also identified by MboI restriction enzyme analysis, was found in both patient and his father but not in any other unaffected family member or 50 unrelated normal individuals. This substitution was reproduced by in vitro site directed mutagenesis of full-length VWF cDNA and transiently expressed in COS-7 cells. The corresponding recombinant VWF protein exhibited the full spectrum of VWF multimers, suggesting that the abnormal multimer seen in the patient results from increased proteolysis. PMID- 11019958 TI - Heterogeneous recognition of beta 2-glycoprotein I by antibodies from antiphospholipid syndrome patients. AB - Beta 2-glycoprotein I plays a pivotal role in the binding of antiphospholipid antibodies to phospholipid in patients with antiphospholipid syndrome. In this study the nature of the epitopes on beta 2-glycoprotein I (beta2-GPI) recognised by sera from antiphospholipid syndrome (APS) patients (n = 15) was investigated and compared to rabbit polyclonal and mouse monoclonal anti-beta2-GPI antibodies. beta2-GPI was only recognised when bound to a high affinity binding support. The antigenic epitope on beta2-GPI recognised by all APS patients was also dependent on disulphide bond integrity. Digestion of beta2-GPI with elastase rapidly destroyed the epitope(s) on beta2-GPI recognised by antibodies in 91% of APS patients. The main cleavage occurred at tryptophan316-lysine317 in the fifth domain. Digestion with staphylococcal V8 protease resulted in a 50% reduction in antibody binding in 81% of patients and the cleavage sites mainly involved the first domain of the molecule. There was considerable variability in the recognition of six different species of beta2-GPI by serum from APS patients. The epitopes on beta2-GPI bound by APS sera appear conformationally determined in all patients but are quite heterogeneous in the regions of beta2-GPI that are recognised. PMID- 11019959 TI - Multi-cellular activation in vivo by endotoxin in humans--limited protection by adenosine infusion. AB - The influence of adenosine infusion (40 microg/kg/min for 4 h) on inflammatory and hemostatic parameters was investigated in healthy males without (n = 10) or with (n = 11) intravenous endotoxin injection (4 ng/kg). Without endotoxin, adenosine elevated circulating leukocytes and circulating platelet-leukocyte aggregates. Endotoxin activated platelets and leukocytes in vivo. Platelet activation was seen as slightly increased platelet P-selectin expression, decreased platelet counts, and elevated plasma soluble P-selectin (from 39.6 +/- 3.4 to 68.9 +/- 6.6 ng/ml, P<0.01). Leukocyte activation was evidenced by increased CD1 lb expression (from MFI of 0.54 +/- 0.02 to 2.21 +/- 0.17; P<0.01) and plasma elastase levels (from 25.3 +/- 2.5 to 169.3 +/- 22.5 ng/ml: P <0.01). Endotoxin also enhanced platelet and leukocyte responsiveness to in vitro stimulation. Endotoxin induced von Willebrand factor secretion (from 92 +/- 8 units to 265 +/- 19 units at 4 h; P <0.001) and enhanced thrombin generation in vivo. Endotoxin induced leukocytosis and thus increased circulating platelet leukocyte, mainly platelet-neutrophil, aggregates. Adenosine caused slight attenuation of platelet reactivity to agonist stimulation, enhanced the endotoxin induced leukocytosis, and detained more platelet-leukocyte aggregates in circulation, but did not attenuate endotoxin-induced neutrophil elastase secretion, von Willebrand factor secretion, or thrombin generation. Thus, endotoxemia induces multi-cellular activation in vivo. Adenosine inhibits leukocyte adhesion and extravasation, and mildly attenuates platelet responsiveness and soluble P-selectin release. Adenosine has the potential of becoming a therapeutic antiinflammatory drug, but an optimal treatment strategy needs to be developed. PMID- 11019960 TI - Functional analyses of patient-derived IgG monoclonal anticardiolipin antibodies using in vivo thrombosis and in vivo microcirculation models. AB - Antiphospholipid antibodies (aPL) have been associated with thrombosis and pregnancy losses in patients diagnosed with antiphospholipid syndrome (APS) and enhance thrombus formation in vivo in mice, but the mechanism of thrombosis by aPL is not completely understood. It has been proposed that aPL may affect endothelial cell (EC) function and/or induce their activation, transforming their anticoagulant surface into procoagulant, thus predisposing to thrombosis. It has been proposed that aPL may affect EC cell function and/or induce their activation, transforming their anticoagulant surface into procoagulant, thus predisposing to thrombosis. This study proposes to test the hypotheses that some IgG anticardiolipins (IgG aCL) with thrombogenic properties in mice, exert their effects through activation of endothelium. We studied seven patient-derived monoclonal aCL for their thrombogenic properties in an in vivo pinch-induced thrombosis model, and their functional activities in activating EC by analyzing in vivo leukocyte adhesion to endothelium in microcirculation in venules in exposed murine cremaster muscle and in vitro adhesion molecule expression in cultured EC. The binding of the monoclonal aCL to EC was also tested. In addition to the previous identified thrombogenic IS2, four of the five new more IgG monoclonal aCL (from two patients) were found to be thrombogenic. Of these five thrombogenic aCL, three caused more in vivo leukocyte adhesion to EC in microcirculation, as compared to that induced by the H2 control human monoclonal IgG, and enhanced expression of adhesion molecules (particularly VCAM-1) on cultured EC. These data show that about 2/3 patient-derived IgG monoclonal aCL are thrombogenic and suggest that some thrombogenic IgG aCL exert their effects through activating EC. PMID- 11019961 TI - Prothrombin activation is increased among asymptomatic carriers of the prothrombin G20210A and factor V Arg506Gln mutations. AB - The risk of venous thrombosis is increased in individuals who carry specific genetic abnormalities in blood coagulation proteins. Among Caucasians, the prothrombin G20210A and factor V Arg506Gln (FV R506Q) mutations are the most prevalent defects identified to date. We evaluated their influence on markers of coagulation activation among participants in the Second Northwick Park Heart Study, which recruited healthy men (aged 50-61 years) from nine general medical practices in England and Wales. They were free of clinical vascular disease and malignancy at the time of recruitment. Genotypes for the two mutations were analyzed using microplate array diagonal gel electrophoresis, and coagulation markers (factor XIIa; activation peptides of factor IX, factor X, and prothrombin; fibrinopeptide A) were measured by immunoassay. Factor VII coagulant activity and factor VIIa levels were determined by a functional clotting assay. Among 1548 men genotyped for both mutations, 28 (1.8%) and 52 (3.4%) were heterozygous for prothrombin G202 IOA and FV R506Q, respectively. The only coagulation marker that was significantly associated with the two mutations was prothrombin activation fragment FI+2 [mean +/- SD, 0.88 +/- 0.32 nmol/L in men with prothrombin G20210A (p = 0.002) and 0.89 +/- 0.30 in men with FV R506Q (p = 0.0001) versus 0.72 +/- 0.24 among non-carriers for either mutationl. This data provides conclusive evidence that heterozygosity for the prothrombin G20210A as well as the FV R506Q mutations in the general population leads to an increased rate of prothrombin activation in vivo. PMID- 11019962 TI - von Willebrand disease in a pediatric-based population--comparison of type 1 diagnostic criteria and use of the PFA-100 and a von Willebrand factor/collagen binding assay. AB - Definitive diagnosis of type 1 von Willebrand Disease (VWD) remains a problem. Provisional consensus guidelines for the diagnosis of definite and possible type 1 VWD were prepared by the Scientific Subcommittee on von Willebrand factor (VWF) of the Scientific and Standardization Committee (SSC) of the International Society on Thrombosis and Haemostasis (ISTH) during the 1996 annual meeting for the specific purpose of further evaluation in retrospective and prospective studies by a Working Party on Diagnostic Criteria (1996 Annual Report of the SSC/ISTH Subcommittee on VWF). In the first phase of this study, we compared 2 definitions of type 1 VWD. each with 3 criteria: significant bleeding history, laboratory investigations, and family history. Using the ISTH consensus guidelines for type 1 VWD definition, significantly fewer patients were diagnosed with definite type 1 disease as compared to our "in house" Hospital for Sick Children (HSC) criteria (4 vs. 31). While we recognize that the provisional ISTH consensus guidelines were not intended for clinical use, we believe that the results of our studies are of interest and will assist in any future refinements to the ISTH guidelines. In the second phase of this study, we investigated the utility of 2 new tests, a laboratory screening test and a functional test, for VWD in our well characterized, pediatric-based population. The Platelet Function Analyzer (PFA-100) provides an in vitro measure of primary hemostasis under conditions of high shear, using disposable cartridges containing collagen and either epinephrine or ADP. All tested subjects with types 2 or 3 VWD had prolonged PFA-100 closure times (CTs) with both cartridge types (n = 17) and prolonged bleeding times (n = 14). In subjects with definite type 1 VWD, 20/24 (83%) had prolonged CTs with the collagen/ADP cartridge (19/24 (79%) with collagen/epinephrine), compared with 7/26 (27%) with prolonged bleeding times. In subjects with definite types 1, 2, or 3 VWD, collagen/ADP CTs were abnormal in 37/41 subjects, giving an overall sensitivity of 90%. With this high sensitivity, the PFA-100 is a better screening test for VWD than the bleeding time. We also tested a VWF collagen-binding assay (VWF:CBA) as a functional test for VWF, in comparison with the more routinely-used ristocetin cofactor assay (VWF:RC0). The VWF:CBA is based on an ELISA technique, which has the potential to be more reproducible than the VWF:RC0. We found that the VWF:CBA detected 43/49 (88%) subjects with definite types 1, 2, or 3 VWD, performing as well as the VWF:RC0, that detected 42/48 (88%). We also showed that, used in conjunction with VWF antigen levels, the VWF:CBA may be useful in classification of VWD subtypes. PMID- 11019963 TI - A comparison of INRs determined with a whole blood prothrombin time device and two international reference preparations for thromboplastin. AB - Oral anticoagulant therapy is usually monitored with the prothrombin time (PT) on citrate plasma samples. In recent years instruments have been developed for measurement of the PT in non-citrated whole blood. In the present study, the manufacturer's calibration of one type of device (CoaguChek) in terms of the international normalized ratio (INR) was evaluated by one laboratory. Three subsequent lots of test strips for the CoaguChek were investigated using blood samples from 56 coumarin-treated patients. Citrated plasma samples from the same patients were analysed with two international reference preparations for thromboplastin (IRP), i.e., rTF/95 (recombinant human) and RBT/90 (rabbit brain). There were statistically significant INR differences between CoaguChek and the international reference preparations (p <0.001), but the mean relative deviation of the INR was not greater than 0.104. Clinically relevant criteria were used to assess the agreement between the CoaguChek and the IRP results. Standard agreement ranged from 82% to 95%. It is concluded that these test strips achieved a clinically acceptable level of accuracy. Further studies of patient management with these strips are justified. PMID- 11019964 TI - Protein S Thr103Asn mutation associated with type II deficiency reproduced in vitro and functionally characterised. AB - Protein S functions as a cofactor to activated protein C (APC) in the degradation of FVa and FVIIIa. In protein S, the thrombin sensitive region (TSR) and the first EGF-like domain are important for expression of the APC cofactor activity. A naturally occurring Thr103Asn (T103N) mutation in the first EGF-like domain of protein S has been associated with functional (type II) protein S deficiency. To elucidate the functional consequences of the T103N mutation, recombinant protein S mutant was expressed in mammalian cells and functionally characterised. The expression level of protein S T103N from transiently transfected COS 1 cells was equal to that of wild type protein S. The mutant protein S and wild type protein S were also expressed in 293 cells after stable transfection, and the recombinant proteins purified. In APTT- and PT-based coagulation assays, the mutant protein demonstrated approximately 50% lower anticoagulant activity as compared to wild type protein S. The functional defect was further investigated in FVa- and FVIIIa degradation assays. The functional defect of mutant protein S was attenuated at increasing concentrations of APC. The results demonstrate the region around residue 103 of protein S to be of functional importance, possibly through a direct interaction with APC. PMID- 11019965 TI - Folate administration increases n-3 polyunsaturated fatty acids in rat plasma and tissue lipids. AB - It has been shown that folic acid deficiency can modify polyunsaturated fatty acid (PUFA) metabolism and thus could contribute to the development of cardiovascular diseases. We have studied the effect of folate administration on fatty acid composition of plasma, platelet, erythrocyte, intestinal and liver lipids. Two groups of rats were daily injected with either saline solution (control group) or 5-methyl-tetrahydrofolate (MTHF) solution (MTHF group) for 15 days. Changes induced by folate treatment on n-6 PUFA were not very significant. Linoleic acid only decreased in liver and intestinal phospholipids of MTHF rats whereas arachidonic acid was unaffected by folate administration. However, n-3 PUFA significantly increased in plasma lipid fractions and in platelet, erythrocyte and intestinal phospholipids. It is well known that n-3 PUFA show antithrombotic properties. Thus, the increase of n-3 PUFA observed after MTHF administration might contribute to the prevention of vascular disorders. PMID- 11019966 TI - Single founder mutation (W380G) in type II protein C deficiency in Finland. AB - The present study investigated the genetic basis for type II protein C deficiency in Finland, where this form has an unusually high incidence. We demonstrated that, first, a single novel mutation W380G in the protein C gene (PROC) explained 25/26 index patients, estimated to represent two thirds of all families with type II deficiency in Finland. Second, extended chromosomal conservation, i.e. a specific haplotype, around the W380G mutation was indicated in unrelated patients. Third, a local geographical origin for the W380G mutation was suggested by genealogical data. These results are in contrast to the heterogeneity in type II protein C deficiency elsewhere, but closely parallel disorders of the Finnish disease heritage. The high frequency of the type II disease can be explained by founder effect and subsequent enrichment of a single mutation in Finland. The present study also provided a simple means for genetic diagnosis of this disease and the genetic test can be included in the routine screenings in this population. PMID- 11019967 TI - Macrophages synthesize factor X and secrete factor X/Xa-containing prothrombinase activity into the surrounding medium. AB - Activation of the coagulation cascade, mediated by various monocyte/macrophage procoagulants, is an important component in the pathology of inflammatory disease. The type of procoagulant expressed may vary between different monocyte/macrophage subtypes and may differ depending on how the cells are treated. In the present study we show that both murine peritoneal macrophages and human adherent synovial cells from rheumatoid arthritis lesions express prothrombinase activity that was inhibited by anti-Factor X antibodies. Northern blot analysis showed that Factor X was transcribed by the murine peritoneal cells and Western blot analysis showed the presence of Factor X antigen. Further experiments showed that the prothrombinase activity was secreted by the cells into the medium in a detergent-sensitive form, suggesting that the prothrombinase is released on small lipid-containing vesicles. PMID- 11019968 TI - Phenotype-genotype correlation in CD36 deficiency types I and II. AB - CD36 deficiency was studied with attention to the phenotype-genotype relationship. The diagnosis of CD36 deficiency was made when CD36 was negative on platelets (type II) or on both platelets and monocytes (type I). Among 827 apparently healthy Japanese volunteers, the type I and II deficiencies were found in 8 (1.0%) and 48 (5.8%), respectively. The T for C substitution at nt478 for Pro90Ser and the insertion of A at nt 1159 constituted the major causes of type I and II deficiencies. The dinucleotide deletion at nt539 had a minor role. In two family studies, we found a previously unreported polymorphic site in the 5' proximal flanking region and the 3'-untranslated region. Including these new polymorphisms, DNA sequence other than the three known mutations affecting CD36 expression was not observed in the CD36 gene, calling into question the previous hypothesis that a platelet-specific silent allele exists near or at the CD36 gene. PMID- 11019969 TI - An alloantibody recognizing the FVIII A1 domain in a patient with CRM reduced haemophilia A due to deletion of a large portion of the A1 domain DNA sequence. AB - We report the development of a FVIII inhibitor in a patient with severe, cross reacting material reduced (CRM(R)) haemophilia A. The level of Factor VIII antigen (FVIII:Ag) measured by ELISA using anti-C2 monoclonal and alloantibodies was 1.9 U/dl. This baseline FVIII:Ag level was increased to 8.3 U/dl after administration of DDAVP. The anti-FVIII inhibitor titer was 2.9 Bethesda U/ml. DNA analysis showed a large deletion of the FVIII gene from exon 4 to 7, corresponding to amino acid residues 111-317 included within the A1 domain. The size of the gene deletion was approximately 28 kb. 5' and 3' breakpoints were identified by sequencing in intron 3 and intron 7, respectively. FVIII mRNA was detected in the patient's peripheral lymphocytes and the deletion spanning exon 4 to 7 was confirmed at the RNA level. Immunoprecipitation experiments using 125I labeled A1, A2 and light chain demonstrated that the inhibitor reacted only with the 54 kDa A1 domain. The inhibitor activity was more than 95% neutralized by A1 domain polypeptide. Our findings suggest a close relationship between the inhibitor epitope and the specific gene deletion with regard to the pathogenesis of the inhibitor in this patient. PMID- 11019970 TI - Hypofibrinogenaemia with compound heterozygosity for two gamma chain mutations - gamma 82 Ala-->Gly and an intron two GT-->AT splice site mutation. AB - We investigated the molecular basis of hypofibrinogenaemia in a woman with a history of recurrent, pregnancy-associated bleeding, and miscarriage. She had a Clauss fibrinogen of 0.9 mg/ml and SDS PAGE of purified fibrinogen showed a normal pattern of chains. However careful inspection of reverse phase chain separation profiles showed apparent homozygosity for a more hydrophilic form of the gamma chain. DNA Sequencing showed only heterozygosity for a CGT-->GGT (Ala- >Gly) mutation at codon gamma82, but further sequencing showed an additional GT splice sequence mutation at the 5' end of intron 2 of the gamma gene. Translation of mRNA containing this intron would result in premature truncation explaining the phenotypic homozygosity of the gamma82 Ala-->Gly substitution. The patient's sister had a mild bleeding disorder with hypofibrinogenaemia and she too was a compound heterozygote for the y mutations. Her nephew had only the novel splice site mutation, while her mother and daughter inherited only the gamma82 Ala-->Gly substitution. PMID- 11019971 TI - Fibrin down-regulates LPS- and PMA-induced tissue factor expression by blood mononuclear cells. AB - Several studies indicate that fibrin may play a functional role in inflammation by modulating a variety of cellular functions. We investigated the effect of fibrin on tissue factor (TF) production by blood mononuclear cells (MNC). Citrated human blood was recalcified and incubated at 37 degrees C for 1-4 h. The resulting clot was lysed by the addition of tissue plasminogen activator (t-PA) and MNC were isolated by density gradient centrifugation. A control blood sample was processed in the same way but omitting calcium addition and clot formation. Clot- and blood-derived MNC did not express detectable TF activity and antigen whatever the incubation time. Clot-derived MNC, however, generated on average 5 fold less TF (activity and antigen) than control cells, when stimulated with lipopolysaccharide (LPS, I microg/ml) for 3 h at 37 degrees C. A reduced TF response of clot-derived cells was also observed at mRNA level as indicated by RT PCR and in situ hybridization. The effect was dependent on the incubation time within the clot, could not be reversed by enhancing LPS concentration or by adding serum, and was maintained if LPS was replaced by the tumor promoter PMA. A reduced TF response was also found when washed MNC were incorporated for 1 h at 37 degrees C within purified fibrin but not when the cells were incubated with fibrinogen, thrombin or fibrin split products alone. indicating that contact with fibrin was responsible for the inhibition of TF production. Fibrin-induced down regulation of TF response to LPS and PMA by MNC may represent a negative feed back aimed at limiting excessive blood clotting activation in immunoinflammatory diseases. PMID- 11019973 TI - The molecular basis for the aberrant production of plasminogen activator inhibitor type 2 in THP-1 monocytes. AB - Plasminogen activator inhibitor type 2 (PAI-2) is a urokinase inhibitor that is expressed primarily in monocytes. THP-1 monocytes, however. contain a unique defect in the production of PAI-2 in that the PAI-2 transcript is truncated and the expressed protein inactive (1). Here we describe the basis of this mutation in THP-1 cells. Southern blot analysis of THP-1-derived genomic DNA indicated that there were no obvious deletions in the structure of the PAI-2 gene. However, assessment of the THP-1-derived PAI-2 transcript by RT-PCR indicated that only exons seven and eight of the normal PAI-2 mRNA could be detected. Cloning of the 5' region of the PAI-2 mRNA by 5-'RACE indicated that the PAI-2 cDNA derived from THP-1 cells is approximately 1329 bp long and contains 180 bp of sequence derived from intron 5. followed by sequences corresponding to exons seven and eight of the normal PAI-2 mRNA. The presence of the intron five fragment in endogenous THP 1 derived PAI-2 mRNA was confirmed by Northern blotting. The absence of any wild type PAI-2 mRNA in these cells suggests that one copy of the PAI-2 allele has been deleted. The remaining allele producing the truncated mRNA appears to have undergone a translocation event and contains a mutation that has disrupted the splicing of the PAI-2 primary transcript. PMID- 11019972 TI - Adenoviral gene transfer of a u-PA receptor-binding plasmin inhibitor and green fluorescent protein: inhibition of migration and visualization of expression. AB - Smooth muscle cell migration plays a role in the development of intimal hyperplasia. Given the established role of the plasminogen activation system in cell migration, an approach to therapy is to overexpress an inhibitor of plasmin. Therefore, an adenoviral vector was constructed encoding the hybrid protein ATF.BPTI, which contains the active domain of bovine pancreas trypsin inhibitor (BPTI), fused to ATF, the amino terminal fragment or receptor-binding domain of u PA. Adenoviral vectors expressing ATF and BPTI individually were also constructed, and a fourth vector was constructed encoding ATF.BPTI linked by an internal ribosomal entry site to Green Fluorescent Protein (ABIG). Both the expression and functionality of the recombinant proteins were established in human vascular smooth muscle cells. Adenoviral gene transfer of ATF.BPTI inhibited SMC migration more efficiently than the expression of ATF or BPTI individually. Expression of ABIG resulted in the co-expression of ATF.BPTI and Green Fluorescent Protein, thereby providing a tool to monitor transfection efficiency and the behavior of the transfected cells. PMID- 11019974 TI - Use of a new monoclonal antibody-based enzyme immunoassay for soluble fibrin to exclude pulmonary embolism. ANTELOPE-Study Group. AB - We prospectively evaluated the diagnostic performance of a new soluble fibrin assay in 303 consecutive patients with suspected pulmonary embolism and examined potentially useful cut-off levels at which this disease can be safely excluded. In addition, the diagnostic accuracy was calculated in the subgroups of in- and outpatients. The ROC curve of the assay in the total study cohort had an area under the curve of 0.69. The cut-off level associated with a sensitivity and negative predictive value of 100% was 20 ng/ml, but the specificity was only 4%. The cut-off level with a sensitivity of 90% was 30 ng/ml, which corresponded with a specificity and negative predictive value of 27% and 86%. respectively. The diagnostic performance was comparable in the subgroups of in- and outpatients. We conclude that the soluble fibrin assay has a low diagnostic accuracy and seems unsuitable as a screening test for the exclusion of pulmonary embolism. PMID- 11019975 TI - Polymorphonuclear leukocyte apoptosis is inhibited by platelet-released mediators, role of TGFbeta-1. AB - Platelets regulate several polymorphonuclear leukocyte (PMN) functions. We have found that thrombin-stimulated platelets potently inhibited PMN apoptosis. Cell free supernatant from increasing concentrations of stimulated platelets inhibited PMN apoptosis in a dose-dependent manner, with an effect similar to that of corresponding concentrations of platelets. At the plateau, platelet supernatant inhibited PMN apoptosis by 54.6 +/- 6.8%, the anti-apoptotic activity being higher than that of GM-CSF and comparable to that of LPS. Neither IL-1ra nor a combination of anti-IL1alpha + betamAb affected the activity of platelet supernatant. In contrast a mAb recognizing the active form of TGF-beta1 significantly decreased this activity. Moreover, exogenous TGF-beta1 inhibited PMN apoptosis in a dose-dependent manner. The active form of this cytokine was indeed present in the supernatant of stimulated platelets at a concentration able to elicit an anti-apoptotic effect. The p38 MAPK inhibitor SB203580 prevented the anti-apoptotic effect of TGF-beta1 in a dose-dependent manner. Interestingly, it also prevented the anti-apoptotic effect of IL-1alpha, but not that of GM-CSF, LPS and dexamethasone. In conclusion, we report for the first time that PMN apoptosis is potently inhibited by platelet-released mediators, that TGF-beta1 mediates an important part of this effect, and that p38 MAPK is involved in the TGF-beta1 signaling leading to its anti-apoptotic effect. These results provide novel evidence to support the central role of platelets in inflammation. PMID- 11019976 TI - Desensitization of the platelet aggregation response to ADP: differential down regulation of the P2Y1 and P2cyc receptors. AB - Platelets activated by ADP become refractory to restimulation, but the mechanism of this process is not well understood. A normal platelet response to ADP requires coactivation of the P2Y(1) receptor responsible for shape change and the P2cyc receptor, responsible for completion and amplification of the response. The aim of the present study was to characterize the desensitization of platelets to ADP and to determine whether or not these two receptors are desensitized simultaneously through identical pathways when platelets become refractory to ADP. It was found that full inhibition of platelet aggregation in response to restimulation by ADP required the presence of ADP in the medium or use of a high concentration (1 mM) of its non-hydrolysable analogue ADPbetaS. Platelets incubated for 1 h at 37 degrees C with 1 mM ADPbetaS and resuspended in Tyrode's buffer containing apyrase displayed a stable refractory state characterized by the inability to aggregate or change shape in response to ADP. ADPbetaS treated platelets loaded with fura-2/AM showed complete blockade of the calcium signal in response to ADP, whereas the capacity of ADP to inhibit PGE1 stimulated cAMP accumulation in these platelets was only diminished. Consequently, serotonin was able to promote ADP induced aggregation through activation of the Gq coupled 5HT(2A) receptor while adrenaline had no such effect. These results suggested that the refractory state of ADPbetaS treated platelets was entirely due to desensitization of the P2Y(1) receptor, the P2cyc receptor remaining functional. Binding studies were performed to determine whether the P2Y(1) and/or P2cyc binding sites were modified in refractory platelets. Using selective P2Y(1) and P2cyc antagonists (A3P5P and AR-C66096 respectively), we could demonstrate that the decrease in [33P]2MeSADP binding sites on refractory platelets corresponded to disappearance of the P2Y(1) sites with no change in the number of P2cyc sites, suggesting internalization of the P2Y(1) receptor. This was confirmed by flow cytometric analysis of Jurkat cells expressing an epitope-tagged P2Y(1) receptor, where ADPbetaS treatment resulted in complete loss of the receptor from the cell surface. We conclude that the P2Y(1) and P2cyc receptors are differently regulated during platelet activation. PMID- 11019977 TI - GPIIb-IIIa antagonist-induced reduction in platelet surface factor V/Va binding and phosphatidylserine expression in whole blood. AB - In addition to inhibition of platelet aggregation, GPIIb-IIIa antagonists may reduce thrombotic events via other mechanisms. In a novel whole blood flow cytometric system, we investigated the effects of GPIIb-IIIa antagonists, in the presence or absence of thrombin inhibitors, on platelet surface-bound factor V/Va and platelet surface phospholipids. Diluted venous blood was incubated with either buffer or a GPIIb-IIIa antagonist (abciximab, tirofiban, or eptifibatide). Some samples were pre-incubated with clinically relevant concentrations of unfractionated heparin (UFH), a low molecular weight heparin, a direct thrombin inhibitor, or buffer only. Platelets were then activated and labeled with mAb V237 (factor V/Va-specific) or annexin V (binds phosphatidylserine), fixed, and analyzed by flow cytometry. In the absence of thrombin inhibitors, GPIIb-IIIa antagonists (especially abciximab) significantly reduced agonist-induced platelet procoagulant activity, as determined by reduced binding of V237 and annexin V. At high pharmacologic concentrations, unfractionated heparin and enoxaparin, but not hirudin, further reduced factor V/Va binding to the surface of activated platelets in the presence of GPIIb-IIa antagonists. Agonist-induced platelet procoagulant activity was reduced in a patient with Glanzmann's thrombasthenia. We conclude that GPIIb-IIIa antagonists reduce platelet procoagulant activity in whole blood and heparin and enoxaparin augment this reduction. Fibrinogen binding to GPIIb-IIIa is important in the generation of platelet procoagulant activity. PMID- 11019978 TI - Factor Xa acts as a PDGF-independent mitogen in human vascular smooth muscle cells. AB - This study investigates the mitogenic effect of the coagulation factor Xa in smooth muscle cells (SMC) from human saphenous vein and the procoagulant activity of these cells. Factor Xa elicited a concentration-dependent increase in [3H]thymidine incorporation. This mitogenic effect of factor Xa was inhibited by DX-9065a and BABCH, indicating the requirement of proteolytic activity of the enzyme. Factor Xa activated the MAP kinases ERK1/2 concentration- and time dependently. PDGF-neutralizing antibodies neither inhibited the increase in [3H]thymidine incorporation nor ERK-1/2 phosphorylation in factor Xa-stimulated cells, suggesting that factor Xa-induced signaling and mitogenic activity in human venous SMC are independent of PDGF. Exposure of SMC to recalcified plasma resulted in a significant thrombin generation which was inhibited by anti-tissue factor antibody, tissue factor pathway inhibitor, inactivated factor VIIa and DX 9065a. These data indicate that interaction of SMC with the clotting system may contribute to venous graft disease, i.e. thrombus formation and intimal hyperplasia. PMID- 11019979 TI - Reduction of von Willebrand factor by endothelial cells. AB - The haemostatic activity of plasma von Willebrand factor (vWF) is a function of multimer size. Only the large vWF multimers are effective in promoting platelet adhesion to a site of vascular injury. We observed that the conditioned medium of cultured human umbilical vein, human microvascular and bovine aortic endothelial cells contained an activity which reduced the average multimer size of plasma or purified vWF. The average multimer size of vWF produced endogenously by human umbilical vein endothelial cells was similarly reduced following secretion. The reducing activity was ablated by pre-treatment with heat or the thiol blocking agents. iodoacetamide, N-ethylmaleimide or E-64, but not by a range of specific serine-, cysteine-, aspartic-, or metalloproteinase inhibitors. Reduction in vWF multimer size was associated with formation of new thiols in vWF and there was no evidence for additional proteolytic processing of vWF. The reducing activity was associated with a protein with an anionic pi that binds heparin and contains reactive thiol(s). These results suggested that the interchain disulfide bonds that link the vWF homodimers near the N-termini were being reduced by a vWF reductase secreted by endothelial cells. In support of this hypothesis, incubation of vWF with the protein reductants, protein disulfide isomerase and thioredoxin, resulted in formation of new thiols in vWF and reduction in the average multimer size of vWF. These findings may have consequences for control of vWF haemostatic activity. PMID- 11019980 TI - Influence of fluvastatin on levels of von Willebrand factor in cardiac transplant recipients. PMID- 11019981 TI - Prevalence of clotting factor deficiencies in a large population with von Willebrand disease. PMID- 11019982 TI - Human herpesvirus 8 infection in haemophiliacs. PMID- 11019983 TI - Thromboplastin sensitivity in waveform analysis. PMID- 11019984 TI - No association of TGF-beta1 antigen with the development of restenosis in patients undergoing peripheral angioplasty. PMID- 11019985 TI - Detection of unexpected lupus anticoagulant during preparation of normal pooled plasma (NPP) from healthy volunteers. PMID- 11019986 TI - Tissue factor expression in monocytes: in vitro compared to ex vivo. PMID- 11019987 TI - rFVIIa--its thrombogenicity. PMID- 11019988 TI - Association of human liver bilirubin UDP-glucuronyltransferase activity with a polymorphism in the promoter region of the UGT1A1 gene. AB - BACKGROUND/AIMS: Gilbert's syndrome is a benign form of a deficiency in bilirubin glucuronidation. It is associated with a homozygous polymorphism, A(TA)7TAA instead of A(TA)6TAA, in the TATA-box of the promoter region of the bilirubin UDP glucuronyltransferase gene. In this study the correlation between this promoter region polymorphism and in vitro human liver bilirubin UDP-glucuronyltransferase enzyme activity was investigated. METHODS: Liver samples from organ transplant donors n=39) and two known Gilbert's syndrome patients were used for measuring bilirubin UDP-glucuronyltransferase enzyme activity and for isolation of DNA followed by detection of the promoter region polymorphism by polymerase chain reaction. Genotypes were assigned as follows; 6/6: homozygous for the A(TA)6TAA allele, 7/7: homozygous for the A(TA)7TAA-allele, and 6/7: heterozygous with one of each alleles. RESULTS: Seventeen out of 39 subjects (44%) had the homozygous 6/6 genotype, 18 subjects (46%) had the heterozygous 6/7 genotype, whereas four individuals (10%) and the two individuals with Gilbert's syndrome had the 7/7 genotype correlated with Gilbert's syndrome. This resulted in an allele frequency of 0.33 for the A(TA)7TAA-allele. The median bilirubin UDP-glucuronyltransferase enzyme activity of the 17 subjects with the 6/6 genotype (1565 nmol/g liver/h) was significantly higher than the activity of the 18 subjects with the 6/7 genotype (985 nmol/g liver/h; p<0.05) and the six individuals with the 7/7 genotype (749 nmol/g liver/h; p<0.005). No significant differences in enzyme activity were found between the 6/7 and the 7/7 genotype groups. CONCLUSIONS: The results indicate a close association between the promoter region genotype and the expression of hepatic bilirubin UDP-glucuronyltransferase enzyme activity. Subjects who have a 7/7 genotype have the lowest enzyme activity, whereas subjects with the heterozygous 6/7 genotype have an intermediate enzyme activity. PMID- 11019989 TI - Composition and immunofluorescence studies of biliary "sludge" in patients with cholesterol or mixed gallstones. AB - BACKGROUND/AIMS: Gallbladder bile from patients with cholesterol or mixed gallstones frequently contains biliary "sludge", a suspension of cholesterol monohydrate crystals and pigment granules embedded in mucin and proteins. The composition of biliary "sludge" and the preferential localization of mucin and proteins could be an indicator for its potential role in gallstone formation. METHODS: Ultracentrifugation (100000 g/l h) was used to precipitate "sludge" from bile, and the concentration difference of its main components between native bile and ultracentrifuged bile samples was calculated. After purification of the sediment, immunolocalization was performed for the detection of mucin, IgA, albumin, aminopeptidase, and anionic polypeptide fraction using polyclonal and monoclonal antibodies. RESULTS: The amount of sludge in gallbladder bile was 4.26 mg/ml-0.78 (mean+/-SEM) in patients with cholesterol and 2.51 mg/ml+/-0.39 in patients with mixed stones and cholesterol was the main component (48.9+/-4.6% and 44.4+/-7.1%). The sediment appeared as a mixture of vesicular aggregates and pigment particles which were linked by a gel matrix of mucin containing cholesterol crystals. While anionic polypeptide fraction and aminopeptidase were associated to pigments, IgA was uniformly spread in the crystalline parts of "core-like" structures, and albumin, when it was present, appeared as randomly located small spots. CONCLUSIONS: Our study demonstrates that the cholesterol content and the distribution pattern of mucin and different proteins is similar in the sediments of biliary "sludge" to that previously shown in cholesterol and mixed gallstones. This suggests that biliary "sludge" represents an early stage of gallstone formation in these patients. PMID- 11019990 TI - A study of fatty liver disease and plasma lipoproteins in a kindred with familial hypobetalipoproteinemia due to a novel truncated form of apolipoprotein B (APO B 54.5). AB - BACKGROUND/AIMS: Familial hypobetalipoproteinemia (FHBL) is a co-dominant disorder characterized by reduced plasma levels of low-density lipoproteins. It can be caused by mutations in the gene encoding apolipoprotein B-100 (apo B), leading to the formation of truncated apo Bs which have a reduced capacity to export lipids from the hepatocytes as lipoprotein constituents. Case reports suggest the occurrence of liver disease in FHBL, but there are no studies of liver involvement in FHBL with defined apo B gene mutations. The presence of fatty liver disease was investigated in a large FHBL kindred. METHODS: Plasma lipoprotein and apolipoprotein analysis, liver function tests, and apo B gene sequence were performed in 16 members of a FHBL kindred. The presence of fatty liver was assessed by ultrasound and computed tomography scanning. RESULTS: The proband, a non-obese heavy drinker male with hypobetalipoproteinemia, had steatohepatitis with fibrosis. He was heterozygous for a novel non-sense mutation of apo B gene producing a truncated apo B of 2745 amino acids (designated apo B 54.5, having half the size of normal apo B-100). Seven other members of his kindred carried apo B-54.5. Although all of them were hypolipidemic, their lipid levels showed a large inter-individual variability not accounted for by polymorphisms of genes involved in apo B metabolism. Four carriers (two heavy drinkers and two teetotallers), irrespective of their plasma lipid levels, had ultrasonographic evidence of fatty liver. In the other four carriers no evidence of fatty liver was found. CONCLUSIONS: In this kindred apo B-54.5 predisposes to fatty liver, which however may require some additional factors to become clinically relevant. PMID- 11019991 TI - Mycophenolate mofetil for maintenance of remission in autoimmune hepatitis in patients resistant to or intolerant of azathioprine. AB - BACKGROUND/AIM: Azathioprine is standard therapy for maintenance of remission in patients with autoimmune hepatitis. However, approximately 15% of patients are intolerant of therapy and 10% do not respond to it. There is a need for alternative therapies. We describe here the results of mycophenolate mofetil therapy in patients with autoimmune hepatitis. PATIENTS: We studied seven patients with type 1 AIH (six female). Three were intolerant of azathioprine and had elevated transaminases and liver histology showing active disease despite prednisolone therapy. Four had been on a dose of 2 mg per kg of azathioprine without complete normalisation of ALT, and had liver biopsies showing active disease. All were treated with mycophenolate 1 g bd and were followed for a median of 46 months (21-59). End points were improvement in histological inflammation, ALT and prednisolone dose. RESULTS: Five of the seven (71%) patients had normal transaminases after 3 months of treatment. The steroid dose fell from a median of 20 mg per day to 2 mg per day at 9 months (p=0.0001) and the hepatic activity index fell from median 11 to 3 (p=0.001) after 7 months of therapy. One patient required dose reduction because of a fall in white cell count. No other adverse effects were seen. CONCLUSIONS: Mycophenolate mofetil is effective and well tolerated in patients with type 1 AIH who are intolerant of, or do not respond to, azathioprine. PMID- 11019992 TI - Effects of lipopolysaccharide on TNF-alpha production, hepatic NOS2 activity, and hepatic toxicity in rats with cirrhosis. AB - BACKGROUND/AIMS: Septic shock results in high mortality in patients with cirrhosis. Nitric oxide synthase 2 (NOS2) is induced by bacterial lipopolysaccharides (LPS) and plays a major role in the inflammatory response to bacterial infections. Little is known about the regulation of NOS2 in cirrhosis under septic conditions. Thus, the aim of this study was to determine tissue NOS2 activity, serum nitrate and tumor necrosis factor (TNF-alpha) levels and hepatic toxicity in cirrhotic rats after LPS administration. METHODS: Serum nitrates, TNF alpha and transaminases were determined after LPS-administration in rats with secondary biliary cirrhosis and in sham-operated rats. Liver, lung, aortic and peritoneal macrophage NOS2 activities were determined by converting L[14C] arginine into L[14C] citrulline in a calcium free medium. Nitrate and TNF-alpha production were determined in a culture medium of peritoneal macrophages after in vivo LPS administration. RESULTS: LPS (1.5 mg/kg) induced 50% mortality in cirrhotic rats and no mortality in sham-operated rats. After LPS, TNF-alpha, nitrate and transaminase levels were significantly higher in cirrhotic rats compared to sham-operated rats. After LPS administration, there were no differences in NOS2 activity in the aorta, lungs, or peritoneal macrophages of the two groups, whereas NOS2 activity was significantly higher in the cirrhotic liver compared to the normal liver. CONCLUSIONS: In rats with cirrhosis, LPS administration induces higher mortality, hepatic toxicity, hepatic NOS2 activation and TNF-alpha release than in sham-operated rats. These results confirm the harmful role of septic shock in liver disease. PMID- 11019993 TI - Diagnosis of small intestinal bacterial overgrowth in patients with cirrhosis of the liver: poor performance of the glucose breath hydrogen test. AB - BACKGROUND/AIMS: Small intestinal bacterial overgrowth is known to occur in association with cirrhosis of the liver and studies are needed to assess its pathophysiological role. The glucose breath hydrogen test as an indirect test for small intestinal bacterial overgrowth has been applied to patients with cirrhosis but has not yet been validated against quantitative culture of jejunal secretion in this particular patient population. METHODS: Forty patients with cirrhosis underwent glucose breath hydrogen test and jejunoscopy. Jejunal secretions were cultivated quantitatively for aerobe and anaerobe microorganisms. RESULTS: Small intestinal bacterial overgrowth was detected by culture of jejunal aspirates in 73% of patients, being associated with age and the administration of acid suppressive therapy. The glucose breath hydrogen test correlated poorly with culture results, sensitivity and specificity ranging from 27%-52% and 36%-80%, respectively. CONCLUSIONS: In patients with cirrhosis, the glucose breath hydrogen test correlates poorly with the diagnostic gold standard for small intestinal bacterial overgrowth. Until other non-invasive tests have been validated, studies addressing the role of small intestinal bacterial overgrowth in patients with cirrhosis should resort to microbiological culture of jejunal secretions. PMID- 11019994 TI - Effects of hepatocellular iron imbalance on nitric oxide and reactive oxygen intermediates production in a model of sepsis. AB - BACKGROUND/AIMS: In mammals iron homeostasis is most important, as imbalance of iron such as iron overload may lead to severe diseases. Recently, it has been shown that the iron regulatory protein-1 is partially controlled by nitric oxide and reactive oxygen intermediates, molecules frequently seen in inflammatory events. The aim of the present study was to investigate the effects of impaired iron homeostasis on the interaction of nitric oxide, and reactive oxygen intermediate formation in hepatocytes in a model of acute inflammation. METHODS: Hepatocytes isolated from Corynebacterium parvum (C parvum)-injected rats were used to examine the formation of nitrogen and oxygen intermediates by iron deprivation and iron overload in the presence of lipopolysaccharide. In addition, we investigated the RNA binding and aconitase activity of iron regulatory protein 1. RESULTS: In the present study we show that iron overload in lipopolysaccharide treated C. parvum-primed hepatocytes downregulated the RNA binding of iron regulatory protein-1 and aconitase activity. Subsequently, we observed a reduced formation of nitrite/nitrate and S-nitrosothiols but an increased production of reactive oxygen species, and hepatocellular damage. Moreover, the addition of iron to cell cultures caused a further increase in cellular damage, a drop in the cellular glutathione pool, and an increase in peroxynitrite and hydroxyl-like radicals. In contrast, addition of deferoxamine (an iron chelator) to lipopolysaccharide-treated C. parvum-primed hepatocytes protected cells by stabilizing the GSH content, maintaining the nitric oxide formation, and by reducing Fenton oxidants. CONCLUSIONS: Our results show that the antioxidative effects of iron chelators prevent the formation of toxic Fenton oxidants in severe inflammatory events, which should be considered in the treatment of disorders characterized by an iron imbalance. PMID- 11019996 TI - Induction of type 1 plasminogen activator inhibitor in human liver ischemia and reperfusion. AB - BACKGROUND/AIMS: The detailed role of type 1 plasminogen activator inhibitor (PAI 1) in liver with ischemia reperfusion injury remains unclear. The aim of the present study was to examine this mechanism, focusing on inflammatory cytokines. METHODS: Eighteen patients who underwent partial hepatectomy (PH) for metastatic liver tumors using only Pringle's maneuver were enrolled in the study. Peripheral blood was taken perioperatively and PAI-1, tissue type plasminogen activator (tPA), plasma aspartate aminotransferase (AST) and cytokine levels were measured. Two liver specimens were taken from each patient, one before ischemia and the other after PH, for detection of mRNA of PAI-1, tPA, tumor growth factor (TGF) beta1, interleukin-1beta and tumor necrosis factor-a. Immunohistochemistry and Western blotting were performed to detect PAI-1 protein in the liver. RESULTS: The average plasma PAI-1 antigen level reached a maximum after the final clamp (121.6+/-5.9 ng/ml). The average PAI-1 level in hepatic veins (140.4+/-6.3 ng/ml) after the last Pringle's maneuver was higher than that in peripheral veins (p<0.0001). A correlation was observed between the ratio of the corrected fluorescent activity of PAI-1 before and after ischemia and TGF-beta1 mRNA levels in the liver after PH (p=0.003). PAI-1 protein could be detected in parenchymal and non-parenchymal cells of the liver obtained after ischemia reperfusion injury. CONCLUSIONS: PAI-1 was produced in human livers after ischemia reperfusion injury provoked by Pringle's maneuver, giving valuable information regarding the degree of warm ischemic damage. PMID- 11019995 TI - Transcriptional activation of heme oxygenase-1 and its functional significance in acetaminophen-induced hepatitis and hepatocellular injury in the rat. AB - BACKGROUND/AIM: Glutathione depletion contributes to acetaminophen hepatotoxicity and is known to induce the oxidative stress reactant heme oxygenase-1. The metabolites of the heme oxygenase pathway, biliverdin, carbon monoxide, and iron may modulate acetaminophen toxicity. The aim of this study was to assess cell type specific expression of heme oxygenase-1 and its impact on liver injury and microcirculatory disturbances in a model of acetaminophen-induced hepatitis. METHODS: Gene expression of heme oxygenase-1 was studied by Northern- and Western analysis as well as immunohistochemistry. The time course of heme oxygenase-1 and -2, cytokine-induced neutrophil chemoattractant-1, and intercellular adhesion molecule-1 was studied by Northern analysis. DNA-binding activity of nuclear factor-kappaB was determined by electrophoretic mobility shift assay. Sinusoidal perfusion and leukocyte-endothelial interactions were assessed by intravital microscopy. RESULTS: Acetaminophen caused a moderate sinusoidal perfusion failure (-15%) and infiltration of neutrophils along with activation of nuclear factor kappaB and intercellular adhesion molecule-1 and cytokine-induced neutrophil chemoattractant-1 mRNAs. Induction of heme oxygenase-1 mRNA and protein (approximately 30-fold) in hepatocytes and non-parenchymal cells paralleled the inflammatory response. Blockade of heme oxygenase activity with tin protoporphyrin-IX abrogated acetaminophen-induced hepatic neutrophil accumulation and nuclear factor-kappaB activation, but failed to affect sinusoidal perfusion and liver injury. CONCLUSIONS: The inflammatory response associated with acetaminophen hepatotoxicity is modulated by the parallel induction of the heme oxygenase-1 gene. However, heme oxygenase-1 has no permissive effect on sinusoidal perfusion and does not affect liver injury in this model. These data argue against a central role of nuclear factor-kappaB activation and neutrophil infiltration as perpetuating factors of liver injury in acetaminophen toxicity. PMID- 11019997 TI - Relationship between copper, zinc and metallothionein in hepatocellular carcinoma and its surrounding liver parenchyma. AB - BACKGROUND/AIM: Accumulation of copper (Cu) in hepatocellular carcinoma (HCC), especially in small tumors, is greater than that in the surrounding liver parenchyma. Metallothionein (MT) is considered to be present as Cu-MT, Zn,Cu-MT or Zn-MT. The aim of this study was to determine the presence and localization of Cu-MT and Zn-MT in HCC and surrounding liver parenchyma. METHODS: In 16 HCC patients, surgically resected specimens including HCC and surrounding liver parenchyma were evaluated. RESULTS: The level of Cu present in small HCC (<4 cm in diameter) was significantly greater than that in the surrounding liver parenchyma (p<0.05). However, the level of Cu in large HCC (>4 cm in diameter) was similar to that in the surrounding liver parenchyma. Analysis by Sephadex G 75 gel filtration revealed that the peak fraction due to Cu was identical to that due to MT in 14 (87.5%) of 16 HCC, the peak fraction due to Cu and Zn was identical to that due to MT in 2 (12.5%) HCC, and the peak fraction due to Zn was identical to that of MT in none of 16 HCC. CONCLUSIONS: Accumulation of Cu in small HCC, in which Cu was present as Cu-MT or Zn, Cu-MT, was greater than that in the surrounding liver parenchyma. Cu accumulation and the presence of MT in the liver may be related to carcinogenesis of HCC, because of the similarity of these findings in the experimental data of Long-Evans rats with a cinnamon-like coat color who develop HCC spontaneously. PMID- 11019998 TI - Prevalence and prognostic value of hepatocellular carcinoma in cirrhotic patients presenting with spontaneous bacterial peritonitis. AB - BACKGROUND/AIMS: This study examined the prognostic power of hepatocellular carcinoma in patients presenting an episode of spontaneous bacterial peritonitis treated with 3rd generation cephalosporins or quinolones, and subsequent prophylaxis with norfloxacin until death or transplantation. METHODS: The study comprises the prospective evaluation of 168 consecutive cirrhosis patients presenting an episode of spontaneous bacterial peritonitis. RESULTS: Hepatocellular carcinoma was diagnosed in 35 out of the 168 (20%) patients included in the study (10 single; 25 advanced tumors). Renal impairment developed in 82 patients. Resolution of infection was achieved in 90% of the cases, the hospital survival being 70%. Renal impairment, advanced tumor stage, albumin, and GGT showed independent prognostic value for hospital mortality. At the end of follow-up 101 patients had died, the 1- and 2-year survival being 36% and 31%, respectively. Four variables independently predicted survival: advanced tumor (OR: 3.9; p=0.00001), renal impairment (OR: 2.1; p=0.00001), bilirubin (OR: 1.6; p=0.02) and creatinine (OR: 1.3; p=0.03). Advanced tumor retained independent predictability in patients surviving hospitalization (OR: 7.5; p=0.0001), the 6 month survival being significantly lower in patients with advanced tumor (12% vs 57%, p<0.00001). CONCLUSION: The prevalence of hepatocellular carcinoma in cirrhotic patients with spontaneous bacterial peritonitis is high, and its presence should be actively sought. Advanced tumor impairs both hospital and long term survival, and should be considered in the design of future trials. PMID- 11019999 TI - Rapid detection of genotypes and mutations in the pre-core promoter and the pre core region of hepatitis B virus genome: correlation with viral persistence and disease severity. AB - BACKGROUND/AIMS: We aimed to clarify the clinical relevance of hepatitis B virus pre-core mutant detection in patients with chronic hepatitis B using a newly developed assay. METHODS: Viral genotypes and pre-core mutations were studied in relation to viral persistence and liver disease severity using INNO-LiPA methodology. The study group included 151 patients with chronic hepatitis B, 85 positive for HBeAg (group I) and 66 positive for anti-HBe (group II). RESULTS: The prevalence of viral genotypes in group I was: 64% A, 1% B, 15% C, 19% D, 0% E, 0% F and in group II: 39% A, 0% B, 2% C, 56% D, 2% E, 2% F (p<0.001). The prevalence of mutations at pre-core codon 28 (M2) was lower in group I (5%) than in group II (64%) (p<0.001). The prevalence of pre-core promoter mutations was also lower in group I (21%) than in group II (61%) (p<0.001). M2 mutations were more frequently detected in genotype D than in genotype A (p<0.001), while the other mutations were not influenced by viral genotype. Serum HBV DNA levels were significantly lower in group II versus group I (p<0.001), and in patients with any of the pre-core mutations versus wild-type sequence (p<0.01). Although cirrhosis was more frequent in group II (37%) versus group I (22%) and in patients with either one of the pre-core mutation (31%) versus wild-type sequence (25%), there was no statistical difference in liver severity assessed by ALT levels and Knodell score. CONCLUSION: Pre-core mutants, whose molecular pattern is strongly dependent on viral genotypes, are associated with viral persistence in anti-HBe positive patients with ongoing chronic hepatitis B. The availability of this rapid assay should allow a precise monitoring of viral pre-core mutants during the course of chronic hepatitis B. PMID- 11020000 TI - HCV-core protein accelerates recovery from the insensitivity of liver cells to Fas-mediated apoptosis induced by an injection of anti-Fas antibody in mice. AB - BACKGROUND/AIMS: Hepatitis C virus (HCV) is a major etiologic agent of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. The aim of this study was to elucidate pathological effects of HCV-core protein on liver cells. METHODS: We have generated transgenic mice carrying HCV-core cDNA (Px-core) and pathologically examined livers of Px-core mice. RESULTS: HCV-core protein was detectable in livers from lines 5 (C5) and 8 (C8) of Px-core transgenic mice. Since chronic hepatitis and cirrhosis precede hepatocellular carcinoma in patients with HCV infection, we tried to examine the effect of repetitive injection of a small dose of anti-Fas antibody in the transgenic mice. Surprisingly, an initial injection of anti-Fas antibody induced resistance of liver cells to the second injection of anti-Fas antibody in both Px-core and littermate control mice. The insensitivity of liver cells induced in the control mice continued for more than 24 weeks after the first injection but was broken within 1 week after partial hepatectomy. However, the sensitivity was restored in the Px-core mice within 12 weeks after the injection. CONCLUSION: HCV-core protein in liver cells may affect persistence of Fas-mediated liver cell injury. PMID- 11020001 TI - Evaluation of long-term efficacy of interferon alpha-2b and ribavirin in combination in naive patients with chronic hepatitis C: an Italian multicenter experience. Ribavirin-Interferon in Chronic Hepatitis Italian Group Investigators. AB - BACKGROUND/AIMS: A combination of interferon alpha and ribavirin has been suggested to reach a higher rate of sustained virological response in patients with chronic hepatitis C than monotherapy. In this study we assessed the long term efficacy of this combination therapy in the treatment of selected Italian naive chronic hepatitis C patients compared to interferon alpha monotherapy. METHODS: We enrolled 428 naive patients who were randomly assigned to receive either recombinant interferon alpha-2b and ribavirin for 24 weeks or interferon alpha-2b alone for 48 weeks. The primary end-point of the study was the rate of sustained virological response. Serum HCV RNA levels were determined before treatment; during treatment at weeks 12 and 24 in the patients receiving the combination therapy; at weeks 12, 24, 36 and 48 in the patients receiving monotherapy; and after therapy at weeks 12, 24 and 48 in the patients in both study groups. RESULTS: Sustained virological response was observed in 43% of the patients treated with combination therapy and in 14% of the patients treated with monotherapy. Logistic regression analysis showed that sustained response was associated with the combination therapy, with HCV genotype other than 1b, with an HCV viral load of 3x10(6) copies/ml or less, with an inflammation score of 7 or less, and with an estimated duration of disease of 10 years or less. CONCLUSIONS: A 24-week treatment course with interferon alpha-2b and ribavirin offers a greater chance of sustained virological response compared to treatment with interferon alpha-2b alone for 48 weeks, and may be indicated as initial therapy in such patients. PMID- 11020002 TI - Combined treatment of relapse of chronic hepatitis C with high-dose alpha2b interferon plus ribavirin for 6 or 12 months. AB - BACKGROUND/AIMS: Retreatment of relapses of chronic hepatitis C with a standard regimen of interferon plus ribavirin for 6 months obtains a sustained response in a minority of patients with high viraemia and genotype 1b. We aimed to assess whether increasing the interferon dose and prolonging the time of combined treatment may enhance the effectiveness, and also to evaluate the tolerability, and to identify the determinants of sustained response. METHODS: Fifty subjects with chronic hepatitis C who had relapsed after one or more courses of a interferon monotherapy were randomised to receive alpha2b interferon (6 MU tiw) plus ribavirin (1000-1200 mg daily) for 6 or 12 months. ALT normalisation and serum HCV-RNA clearance at the end of treatment and 6 months after stopping therapy were used as markers for sustained response. RESULTS: End-of-treatment response was achieved in 48 patients (96%) and 27 (54%) had a complete sustained response. Patients treated for 12 months had a higher rate of sustained response (18/25, 72%; 95% C.I. 0.54-0.89) than those treated for 6 months (9/25, 36%; 95% C.I. 0.17-0.55, p=0.01). Twelve months of therapy was significantly more effective for patients with genotype 1b and baseline serum HCV-RNA greater than 450 000 copies/ml (p=0.005). Seven subjects (14%) discontinued treatment because of side effects. Logistic regression analysis showed 12 months of therapy, young age and low pre-treatment serum HCV-RNA to be independent predictors of sustained response. CONCLUSIONS: Relapsers with genotype 1b and high levels of HCV-RNA will benefit from a 12-month course of 6 MU tiw interferon plus ribavirin, while subjects with genotype 1b and low levels of serum HCV-RNA or with genotype other than 1b may be treated for 6 months. PMID- 11020003 TI - Early addition of ribavirin to interferon in chronic hepatitis C not responsive to interferon monotherapy. AB - BACKGROUND/AIM: Persistence of HCV-RNA in serum early in treatment is a strong predictor of failure of alpha-interferon therapy for chronic hepatitis C. Therefore, we compared the efficacy of ribavirin addition to alpha-interferon with a doubling of the dosage of alpha-interferon in case of lack of early virological response to alpha-interferon therapy. METHODS: Sixty patients were administered interferon alpha2b at the dosage of 3 million units 3 times a week. After the first 4 weeks of therapy, serum HCV-RNA was evaluated. The patients with negative HCV-RNA test received the same treatment for a further 11 months, while those with detectable HCV-RNA were randomized to receive either the same dosage of alpha-interferon plus ribavirin (1000 mg/day) or double dosage of alpha interferon (6 million units tiw) for 11 months. We considered sustained response to be the maintenance of normal alanine aminotransferase and negativity at HCV RNA testing until the end of a 6-month post-treatment follow-up. RESULTS: After the first 4 weeks of treatment, 12 (20%) patients showed virological response and 48 patients (80%) remained positive on HCV-RNA testing. Sustained response was observed in 5/12 (42%) patients with early virological response, in 10/24 (42%) patients without early virological response who were administered ribavirin and alpha-interferon, and in only 1/24 (4%) patients who were administered the double dosage of alpha-interferon (p=0.006). CONCLUSIONS: This study shows the efficacy of the addition of ribavirin to alpha-interferon and the lack of efficacy of doubling the dosage of alpha-interferon in patients without clearance of hepatitis C virus early on in treatment. PMID- 11020004 TI - Genotypic succession of mutations of the hepatitis B virus polymerase associated with lamivudine resistance. AB - BACKGROUND/AIMS: Hepatitis B mutant strains of virus emerging during treatment with the nucleoside analog lamivudine are being increasingly recognized. In the majority of lamivudine-resistant isolates the mutations have been reported to occur within the YMDD motif of the viral polymerase, either as a single mutation M552I or as M552V concomitant with L528M. We analyzed the time course and genetic succession pattern during the emergence of lamivudine resistance. METHODS: Seven patients with breakthrough viremia in the setting of chronic hepatitis (n=5) or recurrent HBV after liver transplantation (n=2) were investigated. Pre- and post breakthrough serum samples were evaluated by single- or second-round PCR amplification and sequencing analysis. RESULTS: Genotypic succession of the virus populations was observed to occur from M552I to M552I/L528M (n=2) and from L528M to M552V/L528M (n=1). The double mutations M552I/L528M (n=4) or M552V/L528M (n=2) were found in six out of seven patients, and represented the stable virus populations throughout the follow-up period. Breakthrough viremia was not associated with the single L528M mutation. The mean duration of uninterrupted treatment with lamivudine until breakthrough was 422 days (range 182-642) and was longer in the setting of chronic hepatitis B than in recurrent hepatitis B after liver transplantation. HBV DNA levels after breakthrough were lower than pretreatment levels in the majority of patients with chronic hepatitis but higher after liver transplantation. CONCLUSION: Our observations show that the virus populations conferring resistance to lamivudine can evolve from single to double mutations at amino acid 552 and 528 of the HBV polymerase, and that M552I/ L528M or M552V/L528M seem to be the predominant mutations arising during long-term antiviral therapy with lamivudine. PMID- 11020005 TI - Jaundice, genes and promoters. PMID- 11020006 TI - Mycophenolate mofetil--a new treatment for autoimmune hepatitis? PMID- 11020007 TI - New pieces to the hepatitis C treatment puzzle. PMID- 11020008 TI - EASL International Consensus Conference on Haemochromatosis. PMID- 11020009 TI - Adult syncytial giant cell chronic hepatitis due to herbal remedy. AB - Syncytial giant cell hepatitis is an uncommon form of chronic hepatitis in adults, although it has been reported in association with viral infection (including HCV and HIV infection), drug reactions and autoimmune disorders. We here report a very unusual case of giant cell chronic hepatitis in a 26-old woman, who had been taking a herbal remedy (ISABGOL) for chronic constipation. The presence of viral and metabolic diseases has been excluded; an autoimmune etiology was very unlikely as the autoimmunity test remained negative during the year of follow-up and the total disease score normalized (from 13 to 8) without corticosteroid treatment. The causative role of the herbal remedy was further supported by the spontaneous and dramatic clinical, biochemical and histologic improvement observed following its withdrawal, despite the absence of any treatment. PMID- 11020010 TI - Budd-Chiari syndrome: combination of genetic defects and the use of oral contraceptives leading to hypercoagulability. AB - A young female, who had been in excellent health and had used third-generation oral contraceptives, was admitted to hospital because of abdominal pain and ascites. Budd-Chiari syndrome (BCS) was diagnosed by radiographic and histological examination. Tests for myeloproliferative disease, deficiency of coagulation inhibitors and paroxysmal nocturnal haemoglobinuria were negative. DNA investigation showed a double heterozygous defect: the Arg506Gln mutation in the factor V gene (factor V Leiden) and G20210A nucleotide substitution in the prothrombin gene. This double defect was also found in the patient's father, who had never experienced an episode of venous thromboembolism. Genetic and acquired thrombogenic risk factors are being detected increasingly in patients with BCS. With the discovery of new genetic defects leading to hypercoagulabiulity an increasing number of patients with serious thrombotic manifestations, such as BCS, will exhibit concurrence of hereditary and acquired risk factors for thrombosis. PMID- 11020011 TI - Vanishing graft syndrome after auxiliary liver transplantation. PMID- 11020012 TI - Fatal hepatitis B reinfection after orthotopic liver transplantation in an HBsAg negative patient following withdrawal of lamivudine. PMID- 11020013 TI - Rectification to the article "Involvement of phosphatidylserine and non phospholipid components of the hepatitis B virus envelope in human annexin V binding and in HBV infection in vitro". PMID- 11020014 TI - HFE mutations in insulin resistance-associated hepatic iron overload. PMID- 11020015 TI - The outcome of acute interstitial nephritis: risk factors for the transition from acute to chronic interstitial nephritis. AB - BACKGROUND: Acute interstitial nephritis has been known as a complication of mainly streptococcal infection for nearly a century. With the advent of infection control, it became a complication caused by antibiotics and later by other drugs, which might have changed the outcome. To determine risk factors for the development of chronic renal insufficiency, and thus, the transition from acute to chronic interstitial nephritis, we performed a retrospective study of all cases of acute interstitial nephritis found by reviewing 1,068 renal biopsies from 1968 to 1997. METHODS: Patients with permanent and reversible renal insufficiency after acute interstitial nephritis were compared with respect to the causative event, the symptoms, and the clinical and histological findings. Differences between the groups were calculated by applying bi- and multivariate analysis. RESULTS: Acute interstitial nephritis was found in 6.5% of all biopsies (64 patients with 68 episodes of acute interstitial nephritis); it was infection induced in 10%, idiopathic in 4%, and drug-induced in 85% of the cases (antibiotics in 13 cases, analgesics in 17, non-steroidal anti-inflammatory drugs (NSAIDs) in 16, diuretics in 5, and various other drugs in 7). Renal insufficiency was reversible in 69% and permanent in 31% (12% partially reversible, 19% irreversible). The infection-induced and idiopathic types of acute interstitial nephritis were always reversible. Drug-related acute interstitial nephritis caused permanent renal insufficiency in 36% with a maximum of 56% in NSAID-induced cases. In drug-induced cases, intake of the suspected drug for more than a month prior to diagnosis caused permanent renal insufficiency in 88% and interstitial granuloma in 31%. Multivariate analysis disclosed the following significant features separating the permanent from the reversible renal insufficiency group: patients in the first group had more tubular atrophy in their histology, more chronic use of mixed analgesics and/or NSAIDs, less oliguria or anuria as an acute symptom, fewer antibiotics as causative agents, more interstitial granuloma, more pronounced interstitial cell infiltration in their histology, and more imaging of renal shrinkage. Renal histology had the highest predictive value. CONCLUSION: Today, acute interstitial nephritis is mainly drug-induced. NSAIDs are the most frequent cause of permanent renal insufficiency after acute interstitial nephritis. Clinically, subacute symptoms, a prolonged intake of the suspect drug, and chronic analgesic or NSAID use are related to a more chronic course of interstitial nephritis. In histology, tubular atrophy, interstitial granuloma, and pronounced interstitial cell infiltration indicate chronicity. PMID- 11020016 TI - Acute tubulointerstitial nephritis in 21 Japanese children. AB - PATIENTS AND METHODS: We studied 21 Japanese children aged 1.2 to 14.9 years with biopsy-proven acute tubulointerstitial nephritis (ATIN; 5 drug-induced, 7 infection-related, 3 tubulointerstitial nephritis and uveitis syndrome (TINU), and 6 unclassified) for clinical presentation, laboratory findings, and outcome to clarify the clinical features of the entity. All developed acute renal failure with peak BUN values from 25 to 164 mg/dl (mean 83 mg/dl) and peak serum creatinine values from 1.5 to 15.1 mg/dl (mean 6.5 mg/dl). All the 7 infection related ATIN were associated with Yersinia pseudotuberculosis infection. Four of the 21 patients underwent dialysis therapy for anuria and 1 patient in the unclassified ATIN showed progression to chronic renal failure. RESULTS: In 20 patients, renal insufficiency lasted for 6 to 73 days: 5 drug-induced; median and ranges 25 (12-33) days, 7 infection-related; 15 (6-22) days, 3 TINU; 65 (55-73) days, and 5 unclassified; 24 (6-34) days. Oral steroid therapy was introduced to the 3 TINU patients because of the prolonged renal dysfunction. Although it was effective in all, 2 of them showed a deterioration of clinical symptoms or renal function after reducing the dosage of steroid. CONCLUSION: Consequently, TINU patients required a longer period of time for the improvement of renal function compared to the other etiologies. PMID- 11020017 TI - The characteristic pattern of aminoaciduria in patients with aristolochic acid induced Fanconi syndrome: could iminoaciduria be the hallmark of this syndrome? AB - BACKGROUND: In Japan the patients with Chinese herbs nephropathy (CHN), aristolochic acids-(AAs) associated renal failure, often present Fanconi syndrome. The aim of this study was to investigate the pattern of aminoaciduria in patients with AAs-induced Fanconi syndrome and to clarify whether it is different from other Fanconi syndromes reported in the literature. SUBJECTS AND METHODS: The subjects consisted of 4 patients with Fanconi syndrome due to AAs. We studied biochemical data and urinary excretion of amino acids in the 4 patients. Amino acids in their urine were analyzed by high performance liquid chromatography (HPLC). RESULTS: Three out of 4 patients showed in common very increased excretion ofproline, hydroxyproline and citruline. Last patient showed the very increased levels of proline and valine. Regarding glycine, which is considered to belong to the same group as imino acid and to be shared with high affinity transport system ofproline, there was not very increased excretion. CONCLUSION: These findings suggest that AAs would predominantly affect the low affinity transport system of proline in the brushborder membrane of proximal tubules because the low-affinity system is considered not to be shared with glycine transport. PMID- 11020018 TI - Serum cystatin C as an endogenous parameter of the renal function in patients with normal to moderately impaired kidney function. AB - BACKGROUND: Cystatin C is a proteinase inhibitor with a low molecular weight. The serum levels of cystatin C are mainly dependent on glomerular filtration rate (GFR) making cystatin C an endogenous parameter of GFR. The aim of the study was to elucidate the applicability of serum cystatin C as a parameter of GFR in patients with normal to moderately impaired kidney function and to estimate a reference interval for serum cystatin C. PATIENTS AND METHODS: Forty-six patients (25 males and 21 females) aged 22 to 83 years with various kidney diseases and 250 blood donors (164 males and 86 females) aged 19 to 64 years were included. Cystatin C was measured by an automated particle-enhanced nephelometric immunoassay, serum creatinine by an enzymatic and by Jaffe method, urine creatinine by an enzymatic method, and GFR by 99mTc-DTPA clearance. RESULTS: Serum levels ofcystatin C and creatinine showed increments with decreasing values of 99mTc-DTPA clearance and a linear relationship was found between 99mTc-DTPA clearance and l/serum cystatin C, l/serum creatinine (enzymatic method), and creatinine clearance. Comparison of the non-parametric receiver-operating characteristic (ROC) plots for serum cystatin C (area under the curve (AUC) = 0.996; SE = 0.005), serum creatinine (enzymatic method) (AUC = 0.899; SE = 0.044), serum creatinine (Jaffe method) (AUC = 0.870; SE = 0.051), measured creatinine clearance (AUC = 0.959; SE = 0.025), and estimated creatinine clearance (0.950; SE = 0.029) revealed significant differences for serum cystatin C and serum creatinine (enzymatic and Jaffe method) (p values: 0.03 and 0.01). No significant differences were demonstrated between serum cystatin C and measured and estimated creatinine clearance (p value: 0.14 and 0.12). The non-parametric reference interval for serum cystatin C was calculated to be 0.51-1.02 mg/l (median: 0.79 mg/l; range: 0.33 - 1.07 mg/l). CONCLUSION: Serum cystatin C seems to be a better parameter of GFR than serum creatinine in adults with various types of kidney disease with normal to moderately impaired kidney function. PMID- 11020019 TI - Lp(a) is increased in hemodialysis patients according to the type of dialysis membrane: a 2-year follow-up study. AB - AIMS: Patients with chronic renal failure treated by hemodialysis develop lipoprotein abnormalities that may contribute to their increased risk ofatherosclerosis. This study shows lipid parameter follow-up procedure according to the type of dialysis membrane in an unselected population of 33 hemodialysis patients. PATIENTS AND METHODS: The study included 33 patients with end-stage renal disease and 110 healthy blood bank donors of Tenon Hospital. Cholesterol and triglycerides were determined by enzymatic methods, apoA-I, apoB by immunoturbidimetry and Lp(a) by immunonephelemetry. Apo(a) phenotyping was performed by agarose gel electrophoresis followed by immunoblotting. Patients and controls subjects were estimated by Student's t- and chi2-tests. RESULTS: Patients dialyzed with low-flux membranes had Lp(a) concentrations higher than patients dialyzed with high-flux membranes. Patients dialyzed with polyacrylonitrile membranes (AN 69) had an apoA-I concentration significantly lower than patients dialyzed with hemophane or polysulfone membranes. We also confirmed some of the lipid abnormalities and high Lp(a) concentrations in ESRD patients. CONCLUSION: These results may contribute to a more rational choice of the dialysis membrane in hemodialysis patients. PMID- 11020020 TI - Fracture and bone mineral density in hemodialysis patients. AB - AIM: The aim of this cross-sectional study was to determine in hemodialysis patients the pattern of low trauma fracture, the ability of dual X-ray absorptiometry (DXA) to discriminate between patients with and without fracture, and the magnitude, distribution and mechanism of bone loss. PATIENTS AND METHODS: Eighty-eight patients were studied. Bone mineral density (BMD) was measured by DXA at lumbar spine (LS), femoral neck (FN) and 3 radius sites (UD, MID and 1/3R). In 11 patients (12.5%), 16 fractures occurred and were predominant at the distal forearm and ribs. RESULTS: Patients with fracture had a significatively lower BMD Z-score at LS (-1.34 +/- 1.66 vs -0.42 +/- 1.23), at FN (-1.58 +/- 1.25 vs -0.60 +/- 1.01), at MID radius (-2.59 +/- 1.34 vs -0.93 +/- 1.76) and 1/3 radius (-1.62 +/- 1.60 vs -0.39 +/- 1.32). They also had a longer history of dialysis (113 +/- 64 vs 53 +/- 65 months). Prevalence of osteoporosis varied from 23% at LS to 50% at MID radius. CONCLUSION: Multiple regression analysis showed that there was no influence of gender, age, parathormone status and primary renal disease on BMD. However, at FN, UD, MID and 1/3 radius, a significantly negative correlation was found between length of dialysis and BMD Z-score. By contrast at LS, there was a positive correlation between age at onset of dialysis and BMD Z score. Despite occurrence of fracture at the fistula forearm, BMD levels were similar in both arms. PMID- 11020021 TI - Hemodialysis causes changes in dynamic vectorcardiographic ischemia monitoring parameters. AB - AIMS: The aim of this study was to establish whether changes in parameters reflecting myocardial ischemia QRS vector difference (QRS-VD), ST change vector magnitude (STC-VM) and ST vector magnitude (ST-VM6) during hemodialysis (HD) registered by MIDA (myocardial infarction dynamic analysis) are related to changes in blood volume (BV), extracellular water (ECW) and blood biochemistry. PATIENTS AND METHODS: Fifteen hemodialysis (HD) patients were studied. Computerized vectorcardiography was used for on-line dynamic analysis of ST segment and QRS complex changes. BV changes were monitored non-invasively and continuously with the CRIT-LINE instrument. Bioelectric impedance analysis (BIA) was used for ECW estimation. Blood samples were taken before and after hemodialysis for hemoglobin (B-Hb), hematocrit (B-Hcr), sodium (P-Na), chloride (P-Cl), magnesium (P-Mg), potassium (P-K), ionized calcium (P-iCa), phosphate (P Pi) and astrup measurement. RESULTS: During dialysis treatment QRS-VD, ST-VM6 and STC-VM did not change in parallel. According to the linear mixed model, no statistically significant changes were noted in ST-VM6 during dialysis (time effect p = 0.5635). On the other hand, QRS-VD and STC-VM showed a statistically significant linear trend (time effect for QRS-VD p = 0.0001 and for STC-VM p = 0.0004). Changes in both ECW and BV affected the change in QRS-VD and in STC-VM. CONCLUSION: During HD treatment changes in the vectorcardiographic ischemia monitoring parameters QRS-VD and STC-VM are mostly related to ECW and BV changes and may give a false positive impression of myocardial ischemia. The ST-VM6 trend is less markedly influenced by volume changes. PMID- 11020022 TI - Clinical characteristics associated to atrial fibrillation in chronic hemodialysis patients. AB - BACKGROUND: Atrial fibrillation (AF) is the most common sustained arrhythmia diagnosed in non-uremic patients and its prevalence increases in older subjects, however, information concerning AF in dialysis patients is scarce. Therefore, we carried out a prospective cross-sectional study from September 1996 to December 1996 in order to evaluate the prevalence and some of the clinical characteristics associated to AF in hemodialysis (HD) patients. SUBJECTS AND METHODS: 316 HD patients (age 63 +/- 12 years, dialysis duration 69 +/- 71 months) treated in three different hospital-based units were studied. Standard 12-lead electrocardiograms (ECGs) carried out in the interdialytic day during the study period were reviewed. Data concerning age, history of ischemic heart disease (IHD), cerebrovascular disease (CVD), peripheral vascular disease (PVD), presence of diabetes, smoking history and antihypertensive therapy were collected. Systolic and diastolic blood pressure, fasting cholesterol and triglycerides, albumin and hemoglobin were also derived from the clinical records. Performance status was assessed by Karnofsky index (Ki). RESULTS: 74 patients (23.4%) had persistent AF, i.e. presence of AF in all (at least two) ECGs performed in the study time. Patients with AF were older (age 69 +/- 10 vs 62 +/- 12 years, p < 0.001), had lower Ki (54 +/- 20 vs 68 +/- 17, p < 0.01), cholesterol (182 +/- 46 vs 198 +/- 52 mg/dl, p < 0.01) and albumin (3.9 +/- 0.5 vs 4.1 +/- 0.5 g/dl, p < 0.001) compared to those with no AF. Prevalence of IHD (44.5% vs 19%, p < 0.05) and PVD (23% vs 11%, p < 0.05) was higher among AF patients. Logistic regression analysis showed that IHD (p < 0.001) and Ki (p < 0.01) were independently associated to AF. CONCLUSION: We conclude that AF is a frequent arrhythmia in HD patients treated in hospital-based dialysis units, especially in those with low performance status. It appears to be associated to the atherosclerotic damage of coronary arterial tree. Prospective studies are necessary to assess whether it could contribute to cardiovascular morbidity and mortality in end-stage renal disease. PMID- 11020023 TI - Safety and tolerability of a multidose formulation of epoetin beta in dialysis patients. Collaborative Study Group. AB - AIM: Previous studies in healthy volunteers and renal patients have demonstrated the favorable tolerability of a new multidose formulation of epoetin beta. The aim of this open, multicenter study was to further assess the safety, tolerability and efficacy of this formulation ofepoetin beta in patients with end stage renal disease (ESRD). MATERIALS AND METHODS: 375 adult patients receiving maintenance epoetin therapy for renal anemia were switched to the multidose formulation of epoetin beta for 12 weeks, using the same dosage and route of administration. RESULTS: Adverse events were experienced by 123 patients (33%), most commonly hypertension (5.6%) and hypotension (4.5%). Few patients (2%) were prematurely withdrawn because of tolerability concerns. No clinically relevant changes in blood pressure or laboratory variables were observed. Compared with baseline, hemoglobin and hematocrit values remained essentially unchanged during treatment with this new formulation of epoetin beta. No changes in iron metabolism parameters were apparent, and nearly all patients (94%) did not require blood transfusions during the study. CONCLUSION: The results of this study indicate that the multidose formulation of epoetin beta is safe and well tolerated in patients with ESRD. Moreover, switching patients to this new formulation of epoetin beta does not compromise therapeutic efficacy. PMID- 11020024 TI - Minimal change nephrotic syndrome in a 65-year-old patient following influenza vaccination. PMID- 11020025 TI - Recurrence of rapidly progressive glomerulonephritis probably associated with two different kinds of drugs. AB - We describe a rare case that developed a rapidly progressive glomerulonephritis twice in a 69-year-old man during a course of treatment, first with allopurinol and then with piperacillin. The cessation of each treatment was followed by spontaneous recovery in renal function. A renal biopsy showed crescentic glomerulonephritis with mild tubulointerstitial change and a skin biopsy showed leukocytoclastic vasculitis. This is, to our knowledge, a very rare case of crescentic glomerulonephritis, probably associated with vasculitis during a course of treatment with two different kinds of drugs. PMID- 11020026 TI - A reply to Agarwal. Strategies and feasibility of hypertension control in a prevalent hemodialysis cohort. PMID- 11020027 TI - RPA position on dialysis for non-citizens. Renal Physicians Association. PMID- 11020028 TI - Determination of pravastatin by high performance liquid chromatography. AB - BACKGROUND: Pravastatin is a hydrophilic liver-specific inhibitor of the enzyme 3 hydroxy-3-methylglutaryl coenzyme A reductase. It effectively lowers plasma cholesterol and low-density lipoprotein concentrations in humans. Pharmacokinetic studies of pravastatin have been mostly performed by means of radioactively labelled drug or by measuring plasma concentrations with gas chromatography and mass spectrometry. AIMS OF THE STUDY: Aim of our study was to develop a simple, but reliable method which allows the determination of pravastatin plasma concentrations under clinical routine conditions. SUBJECTS, MATERIALS AND METHODS: Samples were prepared by solid-phase extraction on cyclohexyl bond elut cartridges. Chromatography was carried out on an octyl matrix. Triamcinolone acetonide was used as internal standard. The method was linear within the range of 5 to 200 microg/l pravastatin. The coefficient of variation depended on the pravastatin concentration, but was less than 10% throughout. The pharmacokinetics of pravastatin were determined in healthy individuals. Five healthy subjects received single oral doses of pravastatin (60 mg) and one of these subjects additionally received a dose of 80 mg at three different study days. In all subjects blood was sampled 0, 30, 60, 90, 120, 150, 180, 240 and 300 min after drug intake. RESULTS: Peak plasma concentrations of pravastatin were found between 60 min and 120 min after oral administration of 60 mg and reached values between 37 microg/l and 126 microg/l. The calculated AUCs were between 52 ng/ml x h and 311 ng/ml x h and the corresponding plasma elimination half-life times were between 95 min and 165 min. In all subjects plasma concentrations of pravastatin 5 hours after oral drug administration were near the detection limit of the method (5 microg/l). Intraindividually, there was only little variation in the kinetics of pravastatin. However, marked differences were encountered between the subjects studied. CONCLUSION: The data suggest that the determination of pravastatin plasma concentrations by means of a HPLC system can be used for routine analysis of pravastatin plasma concentrations. The obtained pharmacokinetic data in healthy individuals stand in ample agreement with the results of prior studies in which the concentrations of pravastatin were determined by other more sophisticated methods. PMID- 11020029 TI - Use of a propafenone metabolic ratio as a measure of CYP2D6 activity. AB - AIM: The antiarrythmic drug propafenone is metabolized to its main metabolite by CYP2D6, suggesting that its metabolic ratio may be used for CYP2D6 phenotyping. However, reported ratios obtained from plasma concentrations did not reflect the phenotype. The objective of this paper was to find optimal conditions for plasma sampling based on pharmacokinetic data and to investigate whether propafenone/metabolite ratios reflect the CYP2D6 phenotype. PATIENTS, MATERIALS AND METHODS: The present study was conducted in 14 healthy volunteers phenotyped for CYP2D6 activity by a sparteine test. A single dose of oral propafenone (Profenorm PRO.MED.CS Praha a.s.) was administered, and venous blood samples were taken up to 24 hours thereafter. Propafenone and hydroxypropafenone were measured by HPLC. RESULTS: The individual data for the respective propafenone/metabolite metabolic ratio in plasma samples taken at tmax correlated well with the sparteine metabolic ratio used routinely for CYP2D6 phenotyping. However, when the samples were taken 4 hours after drug intake, the correlation was poor. CONCLUSION: The results indicate a possibility to use the propafenone metabolic ratio for determination of the CYP2D6 phenotype in plasma samples taken at single time point (close to the Cmax, i.e. 2 hours after drug intake). PMID- 11020030 TI - An extract of Petasites hybridus is effective in the prophylaxis of migraine. AB - OBJECTIVE: Migraine is still an unsolved problem. This clinical trial investigates the efficacy and tolerance of Petasites hydridus in the prophylaxis of migraine. METHODS: A randomized, group-parallel, placebo-controlled, double blind clinical study was carried out with a special CO2 extract from the rhizome of Petasites hybridus. Following a 4-week run-in phase, 60 patients received either the special Petasites hybridus extract petadolex or placebo at a dosage of 2 capsules (each capsule contains 25 mg) twice daily over 12 weeks. Outcome variables included the frequency, intensity and duration of migraine attacks as well as any accompanying symptoms. RESULTS: The frequency of migraine attacks decreased by a maximum of 60% compared to the baseline. This reduction in migraine attacks with petadolex was significant (p < 0.05) compared to placebo. No adverse events were reported. Petasites was exceptionally well tolerated. CONCLUSIONS: The results suggest that migraine patients can benefit from prophylactic treatment with this special extract. The combination of high efficacy and excellent tolerance emphasizes the particular value that Petasites hydridus has for the prophylactic treatment of migraine. PMID- 11020031 TI - Effects of ondansetron on portal hemodynamics in liver cirrhosis. AB - OBJECTIVE: The double-blind randomized pilot study was undertaken to compare the effects of a 10-day course of ondansetron 8 mg/day and propranolol 80 mg/day perorally in treating portal hypertension. SUBIECTS AND METHODS: 16 patients with liver disease were enrolled in the study. Measurements of portal vein diameter, portal blood flow velocity and portal blood flow volume were done at days 1, 5 and 10 of treatment using duplex Doppler sonography. RESULTS: The propranolol group demonstrated a decrease in portal venous diameter, while patients treated with ondansetron exhibited reduced portal blood flow velocity values. A decreased portal blood flow volume was found in both groups after 10 days of therapy. CONCLUSION: No statistically significant differences were found between the groups with the exception of portal venous diameter which is significantly lower at the end of the treatment in the case of propranolol. PMID- 11020032 TI - Knowledge of and attitudes to pharmacotherapy in medical inpatients. AB - OBJECTIVE: To analyze different aspects of patients' knowledge and attitudes to pharmacotherapy in medical inpatients. PATIENTS: 183 patients hospitalized in the Department of Medicine of University Hospital "Merkur", Zagreb, Croatia were investigated. METHODS: A questionnaire was designed to investigate patients' knowledge of drugs they were taking before admission to the hospital and drugs they are receiving during hospitalization. Patients were asked to give drug names, dosage and reasons for their prescription. Patients' rating of the importance of some drug characteristics (dosage, indication, precautions, side effects, mode of action) was evaluated. RESULTS: A representative group of patients (mean age 55.5 years, range 17-86, SD 16.1; 89 men, 94 women; 50 hematological, 44 cardiological, 50 gastroenterological and 39 nephrological patients) showed a significantly better (p < 0.000001) overall knowledge of drugs taken prior to admission compared to the knowledge of drugs that they were receiving during hospitalization. Overall drug knowledge did not differ significantly between groups of patients stratified according to gender, ward, number of drugs they were taking or duration of treatment. In older patients (p < 0.0001) and in those with lower education (p < 0.001) a significantly worse overall knowledge was observed. On a 1-5 semiquantitative scale patients rated dosage as the most important and mode of action as the least important drug characteristic (average 3.62 and 2.08, respectively). Of all patients, 94.5% pointed out physicians as one of their sources of drug information, written drug information followed in 40.4% and pharmacists in only 11.5% of patients. CONCLUSIONS: Our results agree with the results of the few similar studies published to date. A need for better health education of patients is underlined and possible ways of providing drug information for patients are discussed. The need for improvement of physician-patient transfer of drug information as well as the need for written drug information tailored according to patients' needs is underlined. PMID- 11020033 TI - Effect of inhaled low molecular weight heparin on methacholine-induced bronchoconstriction. AB - OBJECTIVES: Recent studies have shown that inhaled standard heparin exhibits protection towards various bronchoconstrictor stimuli in asthma including methacholine. Low molecular weight heparins (LMWH) (4000-5000 daltons) have higher bioavailability than standard heparins (12,000-16,000 daltons). It is possible that the anti-asthmatic activity of heparin may be molecular weight dependent. The purpose of the present investigation was to study the effect of LMWH on methacholine-induced bronchoconstriction and to compare the effect of LMWH with that of standard heparin. SUBJECTS: Fifteen subjects (7 male, 8 female, mean age: 33 +/- 13 years, range: 20-65) with mild asthma were studied. METHOD: Methacholine bronchial provocation tests were performed in a single-blind, crossover, randomized order and repeated 45 minutes after placebo or aerosolized standard heparin (1.000 U/kg) or aerosolized LMWH (Enoksaparin, Clexane, 0.8 mg/kg). RESULTS: There was no significant difference in baseline FEV1 values between study days. The standard heparin and enoksaparin inhibited bronchoconstriction induced by methacholine. The geometric mean log PD20 values after placebo, standard heparin, and enoksaparin were 0.24 +/- 0.57 (1.74) mg/ml, 0.79 +/- 0.59 (6.17 mg/ml), 0.76 +/- 0.57 (5.75 mg/ml), respectively (p < 0.0009). Three subjects in standard heparin group and two subjects in enoksaparin group showed increased hyperreactivity, the others showed decreased bronchial hyperreactivity. The degree of protection offered by standard heparin and enoksaparin did not show any statistical difference. CONCLUSIONS: These data suggest that both inhaled LMWH and inhaled standard heparin play inhibitory roles in methacholine-induced bronchoconstriction. PMID- 11020034 TI - Development of a pulmonary phenytoin-associated hypersensitivity reaction despite concomitant dexamethasone and prednisolone administration. AB - OBJECTIVE: This is the report about a very early onset of phenytoin hypersensitivity reaction showing respiratory insufficiency, fever, hepatic and skin reactions in a 68-year-old male patient who was concomitantly treated with high dose dexamethasone/prednisolone because of brain edema. CASE REPORT: The patient developed a hypersensitivity reaction one week after starting a phenytoin treatment, 250 mg daily intravenously, because of a focal epileptic seizure after a transsphenoidal resection of a pituitary tumor. The drug hypersensitivity was diagnosed first clinically and was confirmed by an in vitro rechallenge using the lymphocyte transformation test three months later. Phenytoin itself did not stimulate the lymphocytes' proliferation. The test was performed successfully using human reference sera. CONCLUSIONS: The results suggest that protein-bound reactive drug metabolites which are assumed to occur in the sera may be responsible for the reaction. Corticosteroids did not prevent the reaction and even seem to mask laboratory and clinical findings. Dexamethasone might accelerate the accumulation of potential allergotoxic epoxide metabolites. Both phenytoin and dexamethasone should be withdrawn urgently in the case of suspected hypersensitivity syndrome. PMID- 11020035 TI - Kinetic alteration of theophylline at therapeutic doses. AB - OBJECTIVE: To report a case of a patient diagnosed as having bronchial asthma treated with intravenous infusion of theophylline, who presented an episode of altered theophylline metabolism with a disproportionately increase in serum concentrations together with tachycardia and symptoms of intoxication. CASE SUMMARY: A 51-year-old white woman with a history of bronchial asthma was treated with intravenous infusion of theophylline in an Intensive Care Unit. Bayesian pharmacokinetic approach for dose individualization was done and linear kinetics of theophylline were observed. At therapeutic dose, an alteration of theophylline metabolism had happened, that induced a decrease on the theophylline clearance coinciding with an increase of dose. The serum concentration at this dose was 23.9 microg/ml and this was associated with symptoms compatible with intoxication by theophylline. All factors that could change theophylline elimination were analyzed. A possible drug interaction, hypothyroidism and cardiac failure were ruled out. Clinical hepatic insufficiency was not determined, but an increase of hepatic enzymes was observed. Theophylline infusion was suspended during 5 hours, the serum levels returned to therapeutic values and hepatic enzymes returned to their initial values. DISCUSSION: Episodes of anormal theophylline kinetics may occur, even with doses and serum concentrations considered to be therapeutic, especially in critically ill patients or those whose physical condition has deteriorated. This makes it necessary to carry out therapeutic drug monitoring of theophylline in this group of the population. PMID- 11020036 TI - Effects of mixed starter composition on nisin Z production by lactococcus lactis subsp. lactis biovar. diacetylactis UL 719 during production and ripening of Gouda cheese. AB - A starter culture system that produced both acid and nisin at acceptable rates in milk for manufacture of Gouda cheese was developed using nisin Z-producing L. lactis subsp. lactis biovar. diacetylactis UL 719 (UL 719) and a commercial Flora Danica (FD) starter culture. Different compositions of mixed cultures (0, 0.2, 0.4, 0.6 or 0.8% UL 719 with 1.4% FD) were tested for acidification and nisin Z production in milk after 12 h incubation at 30 degrees C. The 0.6/1.4% combination, selected as the optimal mixture of starter cultures, acidified milk to a suitable pH and produced nisin Z at a high concentration of 512 IU/ml. With this optimal combination, FD numbers of citrate-fermenting and non-fermenting bacteria did not change compared with the control (1.4% FD). However, with 0.8% of L. lactis strain UL 719 and 1.4% of the FD starter culture, the numbers of citrate-fermenting and non-fermenting bacteria in fermented milk decreased compared with those obtained when milk was inoculated with 0.2, 0.4 or 0.6% of UL 719 added to 1.4% FD or control cultures (1.4% FD). Mixed starter culture ratios 0.6/1.4%, 0.4/1.4% and 0.5/1.4% (UL 719/FD) were used to manufacture nisin Z containing Gouda cheese which was ripened up to 45 weeks. The composition of control cheeses made with 1.4% FD, and nisin Z-containing Gouda cheeses were similar with respect to percent moisture, fat, salt and protein. During the ripening period, the cell counts observed were approximately two logs higher in cheese made with the 0.6/1.4% mixed starter culture than in control cheese. In experimental cheese produced with 0.6/1.4% (UL 719/FD) mixed starter culture, nisin activity increased from 256 IU/g at the end of manufacture to a maximum of 512 IU/g after 6 weeks of ripening; the levels then decreased to 128 and 32 IU/g after 27 and 45 weeks of ripening, respectively. In contrast, nisin Z was not detected in experimental cheeses made with 0.4/1.4% or 0.5/1.4% (UL 719/FD) mixed starters. Using an affinity purified anti-nisin polyclonal antibody, anti-rabbit gold-conjugate and transmission electron microscopy, nisin Z was found to be localized in the cheese matrix, in fat globules, in the casein phase and concentrated at the fat-casein interface. After 27 weeks of ripening, nisin Z was detected preferentially in the fat globules of the experimental cheese. PMID- 11020037 TI - Predictions of growth for Listeria monocytogenes and Salmonella during fluctuating temperature. AB - We studied the predictive performance of a dynamic modelling approach, combined with predictions from the Food MicroModel software, applied to the growth of Listeria monocytogenes and Salmonella in pasteurised milk, chicken liver pate and minced chicken, under constant as well as fluctuating temperatures. We found that, in general, the accuracy of a prediction under fluctuation temperature was similar to that under constant temperature. Generally, there was a good agreement between predictions and observations. However, the growth of Listeria monocytogenes in pasteurised milk was inhibited largely by the natural flora present. PMID- 11020038 TI - Volatiles for mycological quality grading of barley grains: determinations using gas chromatography-mass spectrometry and electronic nose. AB - The possibility of using an electronic nose or gas chromatography combined with mass spectrometry (GC-MS) to quantify ergosterol and colony forming units (CFU) of naturally contaminated barley samples was investigated. Each sample was split into three parts for (i) ergosterol and CFU analysis, (ii) measurements with the electronic nose and (iii) identification of volatiles collected on an adsorbent with a GC-MS system. Forty samples were selected after sensory analysis to obtain 10 samples with normal odour and 30 with some degree of off-odour. The data set of volatile compounds and the data collected from the electronic nose were evaluated by multivariate analyse techniques. SIMCA classification (soft independent modelling of class analogy) was used for objective evaluation of the usefulness of the data from the GC-MS or electronic nose measurements for classification of grain samples as normal or with off-odour. The main volatile compounds of grain with normal odour were 2-hexenal, benzaldehyde and nonanal, while 3-octanone, methylheptanone and trimethylbenzene were the main volatile compounds of grain with off-odours. Using data from the electronic nose three samples of 40 were misclassified, while data analysis of the volatile compounds detected with the GC-MS, led to six misclassified samples. Regression models (partial least-squares, PLS) were built to predict ergosterol- and CFU-levels with data from the GC-MS or electronic nose measurements. PLS models based on both GC-MS and electronic nose data could be used to predict the ergosterol levels with high accuracy and with low root mean square error of prediction (RMSEP). CFU values from naturally infected grain could not be predicted with the same degree of confidence. PMID- 11020039 TI - The use of transgenic yeasts expressing a gene encoding a glycosyl-hydrolase as a tool to increase resveratrol content in wine. AB - Resveratrol, a phenolic compound produced in grapes, exhibits properties that may contribute to the reduction of the incidence of coronary heart disease and other human health related processes. Recombinant yeast strains expressing the Aspergillus niger abfB gene encoding an alpha-L-arabinofuranosidase or the Candida molischiana bgIN gene encoding a beta-glucosidase have been used in vinifications as tools to increase the resveratrol content of white wine. Glycosylated resveratrol isomers (trans- and cis-piceid) and free resveratrol isomers (trans- and cis-resveratrol) were detected and quantified in white wines. Wines fermented with the strain expressing BgiN showed an increased amount of total resveratrol derivatives, particularly the non-glycosylated forms. PMID- 11020040 TI - Structural model requirements to describe microbial inactivation during a mild heat treatment. AB - The classical concept of D and z values, established for sterilisation processes, is unable to deal with the typical non-loglinear behaviour of survivor curves occurring during the mild heat treatment of sous vide or cook-chill food products. Structural model requirements are formulated, eliminating immediately some candidate model types. Promising modelling approaches are thoroughly analysed and, if applicable, adapted to the specific needs: two models developed by Casolari (1988), the inactivation model of Sapru et al. (1992), the model of Whiting (1993), the Baranyi and Roberts growth model (1994), the model of Chiruta et al. (1997), the model of Daughtry et al. (1997) and the model of Xiong et al. (1999). A range of experimental data of Bacillus cereus, Yersinia enterocolitica, Escherichia coli O157:H7, Listeria monocytogenes and Lactobacillus sake are used to illustrate the different models' performances. Moreover, a novel modelling approach is developed, fulfilling all formulated structural model requirements, and based on a careful analysis of literature knowledge of the shoulder and tailing phenomenon. Although a thorough insight in the occurrence of shoulders and tails is still lacking from a biochemical point of view, this newly developed model incorporates the possibility of a straightforward interpretation within this framework. PMID- 11020041 TI - Comparison between the growth of Yersinia enterocolitica, listeria monocytogenes, escherichia coli O157:H7 and salmonella spp. in ground beef packed by three commercially used packaging techniques. AB - Growth of the pathogens Yersinia enterocolitica, Listeria monocytogenes, Escherichia coli O157:H7 and strains of Salmonella were compared in ground beef packed in modified atmospheres of 60% CO2/40% N2/0.4% CO (high CO2/low CO mixture), 70% O2/30% CO2 (high O2 mixture) and in chub packs (stuffed in plastic casings). The ground beef was inoculated with rifampicin-resistant or nalidixic acid/streptomycin-resistant strains of the pathogens (final concentration 10(2) - 10(3) bacteria/g) and stored at 4 and 10 degrees C for up to 14 days. At 4 degrees C the shelf life, based on colour stability and background flora development, was prolonged for the high CO2/low CO mixture compared to the two other packaging methods, but at 10 degrees C the shelf life was < 8 days for all the packaging methods. Growth of Y. enterocolitica was nearly totally inhibited both at 4 and 10 degrees C in the high CO2/low CO mixture, while the bacterial numbers in the samples packed in the high O2 mixture increased from about 5 x 10(2) bacteria/g at day 0 to about 10(4) at day 5 at 4 degrees C and to 10(5) at 10 degrees C. Growth in the chub packs was even higher. L. monocytogenes showed very little growth at 4 degrees C in all treatments. At 10 degrees C there was slow growth from about 5 x 10(3) bacteria/g to about 10(4) at day 5 in the high CO2/low CO mixture, while the numbers in the high O2 mixture and the chub packs were about 10 times higher. Growth of E. coli O157:H7 at 10 degrees C in the ground beef was nearly totally inhibited in both the high CO2/low CO mixture and the high O2 mixture. Growth in the chub packs was higher, as the number of bacteria increased 3 log in 5 days. The Salmonella strains (S. typhimurium, S. dublin, S. enteritidis and S. enterica 61:k:1,5,(7)) in the ground beef stored at 10 degrees C for 5 and 7 days grew to a higher number in the high CO2/low CO mixture than in the high O2 mixture. This study shows that the growth of Y. enterocolitica and L. mononcytogenes in ground beef stored in the high CO2 /low CO mixture was not increased as a result of prolonging the shelf life. However, the observed growth of strains of Salmonella at 10 degrees C in this mixture and in chub packs does emphasise the importance of temperature control during storage. PMID- 11020042 TI - Application of foodborne disease outbreak data in the development and maintenance of HACCP systems. AB - Five-hundred and thirty general foodborne outbreaks of food poisoning reported in England and Wales between 1992 and 1996 were reviewed to study their application to the development and maintenance of HACCP systems. Retrospective investigations of foodborne disease outbreaks provided information on aetiological agents, food vehicles and factors that contributed to the outbreaks. Salmonella spp. and foods of animal origin (red meat, poultry and seafood) were most frequently associated with outbreaks during this period. Improper cooking, inadequate storage, cross contamination and use of raw ingredients in the preparation of food were the most common factors contributing to outbreaks. Classification and cross tabulation of surveillance information relating to aetiological agents, food vehicles and contributory factors facilitates hazard analysis. In forming control measures and their corresponding critical limits, this approach focuses monitoring on those aspects that are critical to the safety of the product. Incorporation of epidemiological data in the documentation of HACCP systems provides assurance that the system is based on the best scientific information available. PMID- 11020043 TI - Improvement of the traditional processing and fermentation of African oil bean (Pentaclethra macrophylla Bentham) into a food snack--'ugba'. AB - Inocula for the improvement of the traditional production of 'ugba' were developed by attaching pure cultures of Bacillus subtilis responsible for the natural fermentation of the African oil bean seeds on cowpea granules. Changes in pH, amino-nitrogen and protease activity were used as fermentation indicators. In comparison with the natural fermentation, changes in these process variables were more pronounced. Results also showed that the production time could be significantly reduced. In addition, the overall product quality of 'ugba' produced by the developed inocula was good and highly acceptable. The cultures were stable and viable for over 6 months on the granules of cowpea. PMID- 11020044 TI - Study of the cryotolerance of Lactobacillus acidophilus: effect of culture and freezing conditions on the viability and cellular protein levels. AB - Slow cooling rate and pre-freezing stress brings about a high increase in the cell resistance and preservation of their physiological characteristics. A brutal decrease in temperature (from 37 degrees C to - 80 degrees C) causes a considerable loss of cell viability, in contrast a slow one preserves a survival rate of 75%. Pre-incubation of cells at low temperature (22 degrees C) during 6 h led to the development of cryotolerance indicated by an enhanced capacity to survive after a freezing treatment of 24 h at - 80 degrees C. Exposure of the cells to low pH (5.5) caused a large decrease in cell resistance but did not lead to any significant decrease of survival rate after freezing treatment. However, an increase of 15 +/- 3% in protein level compared to cells cultivated at regulated pH was observed. PMID- 11020045 TI - Gelatinase activities in wounds of healing-impaired mice versus wounds of non healing-impaired mice. AB - The gelatinases, matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9), have been implicated in different aspects of wound repair. However, little is known about MMP-2 and MMP-9 activity in animal models of impaired wound healing. We sought to compare serial gelatinase activities for 25 days after full-thickness excisional wounds in genetically diabetic healing-impaired mice and their nondiabetic non healing-impaired littermates. Wound samples were frozen, homogenized, clarified by centrifugation, and analyzed on zymography gels, and MMP bands were quantitated relative to a conditioned media standard from HT-1080 cells. Gelatinase activity in both diabetic mice and nondiabetic mice increased after the mice were wounded. However, levels of latent gelatinases peaked earlier in the diabetic wounds, and there was more active MMP-2 and MMP-9 in the wounds of the diabetic mice than in the wounds of the nondiabetic mice. Because the higher gelatinase activity in the wounds of the diabetic mice was similar to the higher levels of gelatinase reported in difficult-to-heal wounds such as ulcers and burns, this diabetic mouse model may be useful for studies of these proteinases and their inhibitors in impaired wound healing. PMID- 11020046 TI - Lawn mower-related burns. AB - Lawn mower-related injuries are fairly common and are usually caused by the mower blades. Burns may also be associated with the use of power lawn mowers. We describe 27 lawn mower-related burn injuries of 24 male patients and 3 female patients. Three of the patients with burn injuries were children. Burn sizes ranged from 1% to 99% of the total body surface area (mean, 18.1%). Two of the patients died. The hospital stay ranged from 1 day to 45 days. Twenty-six injuries involved gasoline, which is frequently associated with refueling accidents. Safety measures should involve keeping children away from lawn mowers that are being used. The proper use and storage of gasoline is stressed. PMID- 11020047 TI - Modulation of microvascular permeability by 21-aminosteroids after burn injuries. AB - Burn injuries initiate lipid peroxidation in capillary endothelial cells and cause alterations in microvascular permeability, with subsequent leakage of fluid and protein from the plasma into the interstitium. We evaluated the effects of two lazaroid compounds (U74389F and U75412E) on alterations in microvascular permeability that resulted from burn injuries. A canine model was used for the evaluation of microvascular permeability at the site of the burn injury with the use of a measure of the reflection coefficient (sigma(d)). Hindpaw lymph flow, lymph and plasma total protein concentrations, and arterial, venous, and capillary pressures were measured before burn injuries and for 6 hours in 6 different groups. Footpaw weight gain was then calculated as the percentage of increase of experimental hindpaw relative to the contralateral paw. The damage was attenuated by 20 mg/kg of lazaroid U75412E given before the injuries, but a lower dose was not effective. This agent was also effective in limiting edema formation, as evidenced by changes in footpaw weight gain. However, the administration of either lazaroid compound produced no significant effect on the burn-induced changes in capillary permeability. We conclude that these lazaroids do not prevent burn-induced changes in permeability at the site of injury when administered after an injury. U75412E administered before the injury was effective in limiting the alterations in microvascular permeability. PMID- 11020048 TI - The effect of medical insurance coverage on the obtainment of pressure garments. AB - Pressure garments are used to alter the appearance of immature burn scars. These garments are costly, and delays in obtaining them are frequent. The purpose of the study was to determine the nature of the delays in the obtainment of pressure garments and to examine the role that the payer plays in these delays. The billing and medical records of all patients with burns measured for pressure garments between January 1, 1998, and August 1, 1999, were reviewed. The distribution of payers was as follows: workers' compensation, 37%; state-funded insurance, 32%; health maintenance organizations, 12%; private insurance, 16%; and other, 3%. Payment authorization time for pressure garments was 37 days for state payers and less than 10 days for all other groups. Patients with state funded insurance waited an average of 67 days to receive their garments as opposed to a wait of 20 to 30 days for other payers. The percentage of billed charges paid was least for patients with state-funded and HMO insurance (58% and 51%, respectively). The interval to payment of charges was longer than 60 days for all groups. Marked delays in authorization exist for state-funded reimbursement of pressure garments. Reimbursement for patients with state- and HMO-funded insurance was lower than for other payers. These differences may have an adverse effect on outcome. PMID- 11020049 TI - Exogenous interleukin-10 increased in vitro rejection of allogeneic keratinocytes. AB - Interleukin-10 (IL-10) is an anti-inflammatory cytokine that has been shown to alter HLA expression in some hemopoietic cells. In skin, the presenting capacity of Langerhans' cells is reduced by IL-10, but little is known about the effect of IL-10 on keratinocytes. Using in vitro experiments, we have shown that IL-10 augments the HLA-DR expression of keratinocytes in the presence of interferon gamma. The increase in HLA-DR expression increased the stimulation and proliferation of allogeneic peripheral blood mononuclear cells, which in turn increased the in vitro rejection of allogeneic keratinocytes. Therefore exogenous application of IL-10 to cultured keratinocytes augments the rejection by allogeneic peripheral blood mononuclear cells in vitro conditions rather than having the suspected protective effect. PMID- 11020050 TI - The challenges of children: the first 48 hours. PMID- 11020051 TI - Application of a model to guide ethical decision making in burn treatment. AB - Ethical considerations in the decision to resuscitate and continue treatment of a massive burn injury are complex. As survival improves, the parameters we use in deciding to resuscitate are less clear. The point at which further treatment is futile is often difficult to recognize, and the decision-making process of whether to withhold or withdraw care can cause great anxiety for the family, as well as for the burn team. The identification of the operative ethical principles in this decision-making process helps clarify issues and values. Although there is no simple algorithm for decision making with ethical dilemmas, the systematic application of the decision-making model presented here that consists of 4 prioritized questions that address medical indications, patient preferences, quality-of-life issues, and contextual factors can focus the problem and define options. In addition, early use of a hospital bioethics committee is advocated to resolve conflict and to provide an acceptable forum for discussion. PMID- 11020052 TI - The impact of a burn wound education program and implementation of a clinical pathway on patient outcomes. AB - Patients are often discharged before burn wounds are fully healed, which requires home care nurses to manage complex burn care. This study evaluated patient outcomes at 3 periods: (1) before visiting nurses participated in a burn care education program, (2) after visiting nurses participated in a burn education program, and (3) after the implementation of a burn clinical pathway. A random sample of 54 patient records from the first 2 periods and the entire sample of 12 records from the third period were reviewed. Demographic variables included age, sex, race, percentage of total body surface area burned, and mechanism of burn. Outcome variables measured were weight maintenance, wound infection, pain management, unplanned readmissions to the burn unit, and documentation of patient education. The findings of this study show the positive impact of education in combination with the use of a burn home care clinical pathway. PMID- 11020053 TI - Torchiere-style halogen floor lamps: a need for fire safety awareness. AB - Torchiere-style halogen lamps pose a significant fire hazard. The 500-W halogen bulb in these lamps can generate temperatures as high as 1200 degrees F. Factors related to the design of the lamp also contribute to the fire risk. The Consumer Products Safety Commission has issued various warnings and recalls on these lamps because of a significant number of fire-related incidents and deaths. The impact of these recalls on a random sample of consumers was investigated. A survey of these participants revealed that 60% of lamp owners and nonowners were unaware of the recalls. The majority of respondents were also not aware of the risks of injury, the extreme bulb temperatures, or safety measures designed for these lamps. There is a need for increasing community awareness and education to reduce the fire hazards associated with this product. PMID- 11020054 TI - Nutrition intervention in pediatric patients with thermal injuries who require laparotomy. AB - Increased intra-abdominal pressure is a complication of thermal injuries that is most commonly noted during burn shock or sepsis. Severely elevated intra abdominal pressure requires surgical treatment by laparotomy to avert cardiac, respiratory, and renal compromise. The purpose of this retrospective study was to examine the manipulation of the nutrition program and outcomes in response to such a procedure. Open laparotomy for increased intra-abdominal pressure was necessary for 6 patients admitted to a pediatric burn facility from March 1993 to April 1999. One patient was excluded from the review because he died 2 days after the burn injury (1 day after the laparotomy) and nutrition intervention was not initiated. Four of the five remaining patients received parenteral nutrition within 48 hours of surgery. One patient did not receive parenteral nutrition because the enteral regimen was at the goal by 5 days after the laparotomy. Trophic enteral feedings were initiated in all 5 patients within 48 hours of the operations. Tube feedings were gradually increased and the parenteral nutrition rate was decreased in accordance with gastrointestinal tolerance (abdominal girth, bowel motility). Enteral nutrition was started before abdomen closure in all of the patients. No mechanical, infectious, or mortality-related complications related to the initiation of enteral nutrition after open laparotomies were noted. Surgical intervention by open laparotomy interrupts the postburn nutrition regimen but does not preclude the safe postoperative delivery and advancement of enteral feedings. PMID- 11020055 TI - Psychological principles of burn wound pain in children. II: Treatment applications. AB - The pain involved in acute burn care can be excruciating and intractable. Even the best pharmacologic pain control efforts often fail to adequately control pain, especially procedure-related pain, in pediatric patients with burn injuries. Nonpharmacologic interventions have been found to be effective in reducing pain in both children and adults and can be extremely important adjuvants to standard pharmacologic analgesia in the burn care setting. In the first article in this series, we outlined psychological factors that influence the emotions, cognitions, and behaviors of children during wound care. Building on this theoretical framework, we now present a detailed discussion of the implementation of nonpharmacologic intervention strategies in the burn care setting. Because accurate measurement of discomfort is imperative for the development of interventions and for the evaluation of their efficacy, we begin with a brief review of pain measurement techniques. We follow this with suggestions for tailoring interventions to meet specific patient needs and conclude with a detailed and practical discussion of specific intervention techniques and the implementation of those techniques. PMID- 11020056 TI - Expression of the insulin-like growth factor system and cancer progression in hormone-treated prostate cancer patients. AB - BACKGROUND: The insulin-like growth factor (IGF) system, which is composed of two ligands (IGF-I and IGF-II), two receptors (IGFR-I and IGFR-II) and six binding proteins (IGFBP-1 to -6), plays an important role in cell biology. To identify the main components in the IGF system that contribute to human prostate cancer progression after hormone therapy, mRNA expression of the IGF system in human prostate cancer tissue was systematically examined. METHODS: Expression of the IGF system in 24 carcinomas obtained from total prostatectomies after hormone therapy was examined. Expression levels of mRNA of each component of the IGF system were analyzed by quantitative reverse transcription-polymerase chain reaction procedures and compared with clinicopathologic parameters. RESULTS: Expression of the IGF system, except IGFBP-1, was detected in all prostate cancer tissue samples. The expression of IGF-II and IGFBP-2 significantly correlated with pathologic stage, lymph node metastasis, histologic differentiation and serum prostate-specific antigen (PSA) levels after hormone therapy. However, expression of IGF-I was significantly lower in locally advanced prostate cancer tissue than in tissue from the early stage. Expression of IGFBP-5 was significantly related to only serum PSA levels after hormone therapy. CONCLUSIONS: In the IGF system, IGF-II and IGFBP-2 play a role in human prostate cancer progression and their increased expression is a possible candidate for a prognostic indicator in hormone-treated prostate cancer patients. PMID- 11020057 TI - Characteristics of pediatric urolithiasis in south-east Anatolia. AB - BACKGROUND: Urolithiasis is endemic in Turkey and characteristics of urolithiasis vary in different regions of the world. The aim of the present study was to evaluate the etiological and clinical characteristics and course of pediatric urolithiasis in south-east Turkey. METHODS: The study population consisted of 81 children (52 girls) with urolithiasis at a mean age of 6.2 +/- 4.2 years who were followed up for 1-32 months. RESULTS: Metabolic disorders, anatomical defects and infection stones were found to be the etiological factor in 34.6, 29.6 and 22.2% of patients, respectively, while 13.6% of patients were considered idiopathic. Of all patients, 28.4% were admitted with acute renal failure (ARF) and 72.8% had urinary tract infection. Recurrence was seen in 19.8% of patients at presentation. The localization of the stone was found to be in the upper urinary tract, the lower urinary tract or both in 65.4, 14.8% and 17.3% of patients, respectively. Patients with multiple and bilateral stones had a higher risk for ARF than the others. The risk for chronic renal failure was significantly higher in children with multiple, bilateral or recurrent stones and with ARF at presentation. CONCLUSIONS: Early diagnosis and management of renal stones and urinary tract infections is necessary to prevent the development of ARF or chronic renal failure and to improve the quality of a patient's life. PMID- 11020058 TI - Pathologic examination of renal cell cancer by means of step-sectioning. AB - BACKGROUND: Because incidental detection of small renal cell cancers (RCC) has recently become increasingly common, nephron-sparing surgery for these cases has been more widely performed. Renal cell cancer was investigated by means of step sectioning in order to determine which cases were suitable for nephron-sparing surgery and how it should be performed. METHODS: Pathology specimens obtained from 90 cases of radical nephrectomy were examined in 5 mm sections. We excluded large and invasive tumors from the present study. Particular attention was given to satellite tumor nodules (STN), pseudocapsules of the main tumor and vascular invasion. RESULTS: Satellite tumor nodules of cancer (STN-Ca) were found in seven of 90 cases (7.8%). Tumor size and grade was not correlated with the existence of STN-Ca. The distance between the main tumor and STN-Ca varied. with the pathology of the STN-Ca mostly resembling that of the main tumor. Pseudocapsules of the main tumor were incomplete in 53 cases (58.9%), but extracapsular invasion of more than 1 mm was not seen in tumors less than 50 mm in diameter. Through step sectioning, a further six cases with microscopic vascular invasion were found. Vascular invasion within the main tumor was confirmed in 16 of 17 cases (94.1%) and within normal renal tissue in only one case. CONCLUSIONS: The incidence of existing STN-Ca was not high (7.8%) in the present study, we had not any characteristics in the cases with STN-Ca. As STN-Ca were rarely near the main tumor, we could not expect to resect STN-Ca with main tumor, resection to more than 1 mm outside the pseudocapsule was needed for complete resection of main tumor less than 50 mm in diameter. Normal renal tissue between the tumor and pelvis is a requirement for selecting appropriate cases in nephron-sparing surgery, but we could not remove the risk of leaving STN-Ca completely. PMID- 11020059 TI - Cowper's syringocele causing voiding disturbance in an adult. AB - BACKGROUND: A 21-year-old man presented with a sudden onset of miction pain and post-void dribbling. The diagnosis was Cowper's syringocele due to the findings of retrograde urethrography and urethroscopy. METHODS/RESULTS: The patient underwent transurethral unroofing of the syringocele. The present case is of interest because Cowper's syringocele is usually diagnosed in male infants and children and is rarely found in adults. CONCLUSION: This lesion should be included in the differential diagnosis of voiding disturbances in adults. PMID- 11020060 TI - Congenital urethrocutaneous fistula. AB - BACKGROUND: A 3-year-old boy visited our hospital for aberrance of urination. He had a fistula on his ventral penile shaft. Our diagnosis was congenital urethrocutaneous fistula. METHODS/RESULTS: We performed one-stage repair transverse preputial onlay island flap urethroplasty. Postoperatively, the patient was voiding comfortably with no recurrence of fistula. CONCLUSIONS: Congenital urethrocutaneous fistula is rare. Eighteen cases of congenital urethrocutaneous fistula have been reported previously. We consider the etiology of congenital urethrocutaneous fistula as a deficiency of the urethral plate and fusion of urethral folds. PMID- 11020061 TI - Intrarenal varix mimicking a cystic renal tumor. AB - BACKGROUND: A case of intrarenal varix in a 60-year-old woman is reported. METHODS/RESULTS: The preoperative diagnosis was cystic tumor in the central part of the left kidney. Retrospectively, gadolinium-enhanced magnetic resonance (MR) imaging demonstrated a layered gadolinium sign, indicating the vascular nature of the mass. CONCLUSIONS: Our experience shows that urologists should be aware of the possible presence of a renal cystic mass of vascular origin and of the usefulness of gadolinium-enhanced MR examination for making an identification. PMID- 11020062 TI - Primary squamous cell carcinoma of the prostate. AB - BACKGROUND: Primary squamous cell carcinoma (SCC) is an uncommon tumor of the prostate gland. A 65-year old man complained of obstructive symptoms. METHODS/RESULTS: Transrectal palpation and diagnostic imaging indicated an ordinary adenocarcinoma, although serum prostate-specific antigen (PSA) was normal. Biopsy specimens revealed SCC with the serum SCC antigen elevated. The patient was treated with pelvic irradiation and systemic administration of cis platinum and peplomycin, which resulted in shrinkage of the cancer. CONCLUSION: No evidence of recurrence has been seen for 18 months. PMID- 11020063 TI - Primary malignant melanoma of the male urethra. AB - BACKGROUND: We describe a rare case of malignant melanoma originating from the male urethra. METHODS/RESULTS: This case, which presented a tan-colored lesion at the distal urethra on urethroscopy, was considered as transitional cell carcinoma by transurethral biopsy. Pathologic examination of widely resected urethra revealed scattering cells, which included melanin granules in cytoplasm, and then immunohistochemical stains established the diagnosis as amelanotic melanoma. Although partial penectomy and inguinal lymph node dissection were subsequently performed, 7 months later, total cystourethrectomy was required because of multifocal recurrence at the residual urethra. CONCLUSIONS: In cases of amelanotic melanoma, it should be noted that a lack of melanin granules in tumor cells may lead pathologists and urologists to a misdiagnosis due to the histologic non-specific appearance. With respect to surgical management, of significance is the fact that malignant melanoma may progress multifocally along the total urethra; therefore, total urethrectomy should be considered as radical surgery for the primary site, even if the tumor is confined to the distal urethra. PMID- 11020064 TI - Two-way selection for high and low responsiveness to photostimulation in broiler strain males. AB - Previous studies indicate that responsiveness to photoperiod (PR) has deteriorated in White Rock heavy breed females and males, and that selection for increased PR in females has a positive effect on egg production. In order to determine whether PR can be affected by selection in males, a two-way selection experiment for high and low PR, as expressed through early and late onset of mature semen production under suboptimal photoperiod, was implemented in White Rock heavy breed males. In each selection line, the selected males in each generation were mated to unselected females of the same line. After four generations of selection, males and females of the two selection lines did not differ significantly in 6-wk body weight. Males of the high-PR line, however, achieved mature semen production 18.9 d earlier (P < 0.01) and at a body weight 587 g lighter (P < 0.01) than the males of the low-PR line. Females of the high PR line came into lay 25.9 d earlier (P < 0.01) and at a body weight 375 g lighter (P < 0.01) than females of the low-PR line; they also produced 13.9 more eggs over a 14-wk test period (P < 0.01). These results show that it is possible to affect PR by selection in males, with parallel effects on female PR and reproductive performance. This result carries the potential of improving reproductive performance of male and female broiler chickens through mass selection for early sexual maturity among males. PMID- 11020065 TI - Male broiler breeder fowl display high levels of aggression toward females. AB - Commercial broiler breeder males are reported to display high levels of aggression, often injuring and sometimes killing females. Aggression toward females by mature male domestic fowl is rare and may be regarded as aberrant behavior. The objective of this study was to determine the effects of genetic strain and feed restriction on the sexual and aggressive behavior of male domestic fowl. A 3 x 2 factorial design was used to compare the behavior of three genetic strains (two broiler breeder strains and one commercial laying strain) on two feeding regimens (one restricted and one fed ad libitum) between 25 and 37 d of age. At 22 wk of age, pairs of same-strain males were penned with 20 females of a broiler breeder strain. Behavior was sampled from each pen for six 10-min periods during Weeks 25, 27, 29, 33, and 37. Broiler breeder males displayed significantly more male-to-male (P < 0.0197) and male-to-female aggression (P < 0.0005) than laying strain males. Broiler breeder males chased females (P < 0.0001), forced more copulations (P < 0.0003), and displayed little courtship behavior (P < 0.0001) compared with laying strain males. There were no behavioral differences between broiler breeder strains. It was expected that feed restriction would result in increased aggression. However, males fed ad libitum displayed the most male-to-male (P < 0.0035) and male-to-female (P < 0.0273) aggression. Sexual behavior was not affected by feeding regimen. In conclusion, broiler breeder males display aberrant sexual behavior and extremely high levels of aggression, which are associated with genetic differences, not feed restriction. PMID- 11020066 TI - Egg prices, feed costs, and the decision to molt. AB - On April 7, 1998, the United Poultry Concerns filed a petition with the Department of Health and Human Services of the Food and Drug Administration calling for the elimination of the practice of forced molting of laying hens in the US. In reaction to this petition, this study investigated the economic importance of forced molting as a short-term production management tool for egg producers. The relationship between shell egg prices and feed costs and the occurrence of forced molting in the five shell egg-pricing regions in the US was addressed. The purpose of this analysis was to determine whether forced molting is used to slow egg production during periods of falling or low egg prices or periods of high or rising feed costs. Ordinary least squares was used to test the relationship between the independent variables (egg, corn, and meal prices) and the dependent variable (percentage of layers in molt). In four of the five regions, there was a significant inverse relationship (P < 0.05) between egg prices and the percentage of layers in molt. This analysis suggests that producers were influenced by current egg prices when making the decision to molt. However, the relationship between the percentage of layers in molt and corn and meal prices was less clear. Although a positive relationship between feed prices and molt was found in each region, in only one region was the relationship statistically significant (P < 0.05). PMID- 11020067 TI - Application of the agar gel precipitin test to detect antibodies to Salmonella enterica serovar enteritidis in serum and egg yolks from infected hens. AB - Serological surveillance can be an important component for egg quality assurance programs geared toward controlling problems with Salmonella enterica serovar Enteritidis (S. enteritidis) within a flock. Serum is the primary sample source for the procedures, although egg yolk antibody assays have become popularity in recent years. However, these assays tend to be labor intensive, requiring procedures for extracting antibodies from the yolk followed by assaying the samples. We describe an adaptation of the agar gel precipitin (AGP) test for use in detecting antibodies to S. enteritidis deposited in egg yolks of infected hens. Yolk or sera from infected birds were administered to wells cut into seven well clusters in an agar gel plate, and detection antigen was added to the center well. The agar gels were incubated for 24 h and then examined for the presence of precipitin lines formed by the interaction of antibody with antigen. Three different antigens were tested: S. enteritidis flagella, SEF14 (a 14-kDa fimbrial protein produced ostensibly by S. enteritidis), and a sodium deoxycholate extract of whole S. enteritidis organism. Flagella and the organism extract detected antibodies to S. enteritidis in the yolk and sera, whereas SEF14 was not reactive. Positive reactions were observed in serum 1 wk postchallenge, whereas in yolks, this was further delayed by 1 wk. The sensitivity of the test was slightly less than the standard microagglutination assay, although specificity was slightly higher, as indicated by results from sera and yolks from birds infected with Salmonella enterica serovar Typhimurium. Simplicity and low labor requirements of the assay would allow for the potential testing of several hundred egg samples within a day, which would make up for test shortcomings due to sensitivity. The AGP test could be an important tool for individuals using serological testing to monitor the S. enteritidis situation within their flocks or as a rapid screen for vaccine responses. The assay could also be used in tandem with other AGP tests to screen for the presence of multiple avian pathogens. PMID- 11020068 TI - Effects of treatments with cyclooxygenase inhibitors on chickens infected with Eimeria acervulina. AB - Reduced weight gains and feed conversions are major sources of economic losses from avian coccidiosis. Experiments were conducted to determine possible involvement of up-regulation of cyclooxygenase (COX) activity during coccidia infections that might contribute to reduced weight gain. In a series of trials, young (3 to 5 wk of age) cockerels infected with Eimeria acervulina, a duodenal parasite, received oral treatments with indomethacin (IM), an inhibitor of both COX-1 and COX-2, or nimesulide (NIM), a specific inhibitor of COX-2. Infection doses varied with experiment (from 10(5) to 10(6) oocysts per chick). Effects of infection on weight gain, duodenal lesions, plasma carotenoids, and levels of NO2+ NO3- were determined at 6 d postinoculation. Total oocysts were enumerated in feces collected from 5 through 8 d postinoculation from separate replicate groups. In no trials did treatment with IM reverse effects of infection on weight gain or significantly reduce lesion scores. However, in all trials, IM treatment reduced oocyst output per chick (Trial 1, 15%; Trial 2, 19%; Trial 3, 53%; Trial 4, 29%; Trial 5, 28.5%). Supplementation of feed with 400 and 100 ppm NIM significantly reduced weight gain of, and increased oocyst output from, infected chicks, whereas supplementation with 50 ppm NIM had no beneficial effects on weight gain or lesion scores, but reduced oocyst shedding. The inability of the IM and NIM treatments to reverse infection-associated weight gain suppression suggests that this pathological effect is not linked to increased prostanoid synthesis as a result of COX-2 up-regulation during infection. The inhibitory effects of IM treatment on oocyst shedding suggest that COX-2 products may have immunosuppressive effects in coccidia infection at local sites of infection. The stimulating effects of high-dose NIM treatments on oocyst shedding suggest that this compound may inhibit synthesis of other prostanoids as well. PMID- 11020069 TI - A note on the effects of perches and litter substrate on leg weakness in broiler chickens. AB - Two trials were conducted to investigate the effect of availability of perches on indices of leg weakness in broiler chickens. A third trial investigated the effect of litter substrate on similar indices of leg weakness in broiler chickens. Leg weakness traits examined were walking ability and tibial dyschondroplasia, tibial curvature, foot burn, and hock burn. Body weight was also measured in all trials. The presence of perches in the rearing pens had no effect on any of the indices of leg weakness examined in either trial. There were no consistent effects of perches on BW. Litter substrate significantly affected some indices of leg weakness; birds reared on wheat straw had poorer walking ability and more foot burn than birds reared on wood shavings, and birds reared on hemp waste were intermediate between them. There was no effect of litter substrate on tibial dyschondroplasia or tibial curvature. Turning the straw litter regularly and adding fresh supplies when necessary did not significantly improve indices of leg weakness. It was concluded that wood shavings provide a better litter substrate than straw, but that perches have no beneficial effect on reducing leg weakness in broilers. PMID- 11020070 TI - Adjuvant effects of various lipopeptides and interferon-gamma on the humoral immune response of chickens. AB - The adjuvant effects of various lipopeptides and recombinant chicken interferon gamma (IFN-gamma) on the humoral immune response of laying hens was investigated in four immunization studies. We used the lipopeptide Pam3Cys-Ser-(Lys)4 (PCSL), the conjugate P-Th1 consisting of the lipopeptide P3CS and the T-helper epitope Th1 (FISEAIIHVLHSRHPG), and the conjugate P-Th2 of the lipopeptide P3CSS and the T-helper epitope Th2, which corresponds to the peptide EWEFVNTPPLV, as adjuvants. Human serum albumin (HSA), recombinant bovine somatotropin (RBST), and human immunoglobulin G (IgG) served as antigens in the different experiments. All tested adjuvants enhanced the humoral immune response with various intensities. Chickens showed high antibody titers after the immunization with HSA even without adjuvant, but the adjuvant effects of PCSL and the combination of PCSL and recombinant chicken interferon-gamma (IFN-gamma) were much more pronounced using the antigens RBST and IgG. Especially after the third immunization, higher titers of antibodies were induced by the coadministration of P-Th1 and, to a greater extent, by the combination of PCSL and P-Th1 compared with the use of PCSL. Also, chickens that had received PCSL and P-Th2 showed the highest immune response, even after the second booster. The average concentrations of chicken immunoglobulin Y were significantly higher in 5-mo-old chickens (9.4 mg/mL serum and 10.1 mg/mL egg yolk) compared with 9-mo-old chickens (5.9 mg/mL serum and 5.1 mg/mL egg yolk). The specific serum antibody response was higher in the older chickens than in the younger chickens. Because chicken antibodies are likely to be used increasingly for diagnostic and therapy in the future, lipopeptides and recombinant chicken IFN-gamma may find many applications as adjuvants, thus contributing to the welfare of experimental animals. PMID- 11020071 TI - Alloantigen systems L and P influence phagocytic function independent of the major histocompatability complex (B) in chickens. AB - Synthetic parent stocks were designed to produce progeny among which alleles were simultaneously segregating for nine alloantigen systems, including the MHC (B). Chicks from Ancona-derived B19B19 females crossed with White leghorn B19B21 males were blood typed, resulting in genotypic categories for the A-E, C, D, H, I, L, and P loci with the objective of determining which, if any, of the eight non-MHC alloantigen systems influence or interact with the B system genotypes for blood monocyte phagocytic activity. Leukocytes obtained from whole blood at 2 and 4 wk were separated on a Fico/Lite LymphoH, density gradient and were allowed to adhere to glass coverslips. The resulting adherent monocyte monolayers were incubated with viable Escherichia coli for 1 h and stained with Leukostat, and the phagocytic monocytes and numbers of internalized bacteria per phagocytic monocyte were scored microscopically. The combined results from two separate trials demonstrated that the genotypes of the A-E, C, D, H, and I systems did not differ in the percentage of monocytes exhibiting phagocytosis, whereas significant differences were noted relative to the B system genotype at 2 wk of age (B19B21 > B19B19; P = 0.049), L at 4 wk (L1L1 > L1L2; P = 0.009), and P at 4 wk (P4P4 > P1P1; P = 0.047). The data were further analyzed to determine any interactions of P and L alloantigen genotypes with the B system genotypes; no such interaction was observed. These studies suggest that the L and P non-MHC alloantigen systems have the potential to influence immune responses by modulating phagocytic function in chickens. Furthermore, this modulation seems to be independent of the B (MHC) system. PMID- 11020072 TI - Analysis of chicken bile by gel precipitation reactions using a lectin in the place of antibody. AB - A lectin obtained from black turtle beans (BTB) was precipitated with IgA in chicken bile samples in various forms of agarose gel systems. Ouchterlony-type double-diffusion (ODD) precipitation patterns between the lectin, bile IgA, and heavy chain-specific antibody contained spurs of the type suggestive of partial immunologic identity. The immunoelectrophoresis precipitation patterns between the same three reactants were mirror images and fused on the cathodic side of the immunoelectrophoresis origin. In addition to use in ODD-type gels, BTB could also be incorporated into agarose gels suitable for Mancini (radial immunodiffusion) or Laurell-type rocket electrophoresis. Bile samples obtained from Cornell lines OS and C, broiler breeder males, and University of California-Davis congenic lines were investigated using BTB- and antibody-based methods. The results of this study indicated that IgA was the most frequently detected isotype in bile, occurring in 139 of 156 (89%) samples. Most bile samples (128/156; 82%) also contained IgG, whereas fewer (19/156; 12%) contained IgM. Cornell lines appeared to differ from broiler breeders, having a higher frequency of IgM-positive samples. Of the total bile samples studied, 11% (17/156) of samples from broiler breeders and the Cornell lines appeared to be devoid of IgA; the bile of one broiler breeder was found to be devoid of all three isotypes. Instances were found in which bile samples shown to be negative for IgA by antibody-ODD were shown to be positive by BTB-ODD. Thus BTB appears to be a suitable adjunct to antibody for the study of IgA. PMID- 11020073 TI - Evaluation of normal yellow dent corn and high available phosphorus corn in combination with reduced dietary phosphorus and phytase supplementation for broilers grown to market weights in litter pens. AB - A study was conducted to determine the extent fecal P levels could be reduced while maintaining performance. Various strategies were employed including the use of a high available phosphorus hybrid of corn (HAPC), supplementation with phytase enzyme, and reduced dietary P levels. The use of HAPC resulted in a 50% reduction in phytate-bound dietary P as compared with a normal yellow dent corn (YDC) diet. Dietary nonphytate P was maintained at either NRC (1994) recommendations for appropriate age periods or reduced by 0.075 or 0.15%. Portions of the diets were supplemented with 1,000 units of phytase/kg. Male chicks of a commercial strain were grown to 56 d on the test diets. Broilers fed diets with HAPC had BW, feed conversion, livability, and tibia ash that were equal to or superior to those fed diets with YDC with considerably reduced fecal P content at any dietary level of nonphytate P. Phytase supplementation enabled birds to maintain live performance at lower levels of nonphytate P, further reducing the fecal P output. One of the greatest contributions of phytase was a reduction in mortality at the lower levels of nonphytate P. Dietary P levels could be reduced by 0.075% under NRC (1994) recommendations without adversely affecting live performance; a reduction of 0.15% in conjunction with phytase supplementation maintained BW, feed conversion, and livability but reduced tibia ash. The extent to which dietary P levels can be reduced over the entire feeding program is subject to further research. PMID- 11020074 TI - Limiting amino acids after methionine and lysine with growing turkeys fed low protein diets. AB - The effectiveness of Thr and other amino acids (AA) replacing CP in Met- and Lys adequate diets of Large White male turkeys was studied from 6 wk to market age. Experiment (EXP) 1 examined efficacy of Thr and an AA grouping in corn-soybean (CS) diets containing 82.8 and 77.0% of NRC CP and in corn-soybean-canola-meat (CSCM) diets containing 85.2 and 79.4% of NRC (1984) CP. Experiment 2 compared AA responses in CSCM diets containing 100, 92.5, 85, and 77.5% of NRC (1994) CP. Compared with control CP, 1) 92.5% of NRC CP supported maximum BW, and supplemental Thr or Trp was without effect; 2) 82.8 to 85.2% of NRC CP resulted in reduced BW and breast meat yield (BMY), and supplemental Thr or Trp was ineffective in reversing this reduction, and 3) 77 to 79% of NRC CP resulted in depressed BW and BMY. Supplemental Thr provided a substantial positive BW but no BMY response, whereas a combination of Thr, Ile, Val, Arg, and Trp completely returned BW and partially returned BMY to that of the normal CP control. Turkeys on CS and CSCM assay diet series supported BW responses to CP and AA similarly. We concluded that in low-CP diets containing Met and Lys to requirement, supplemental Thr resulted in improved BW, whereas Thr, Ile, Val, Trp, and Arg returned BW, but not BMY, to normal CP control. PMID- 11020075 TI - Identification of limiting amino acids in methionine- and lysine-supplemented low protein diets for turkeys. AB - Large White male turkeys were fed 100, 85, 70, or 60% of NRC (1994) CP during 7 to 28 d (Experiment (EXP) 1), 8 to 12 wk (EXP 2), and 16 to 20 wk (EXP 3) of age. Diets contained corn, soybean, canola, and meat meals and were supplemented with Met and Lys to requirement. The influence of supplementary amino acids (AA) was studied at each protein level. Turkeys fed 85% CP gained BW similarly to those fed 100% of NRC CP (control) during each age range. Supplemental Thr, Val, and Ile during 7 to 28 d or 8 to 12 wk, or Thr during 16 to 20 wk, did not result in positive BW gain response. For turkeys fed 70% CP, BW gain was depressed compared with the normal-CP control in each period. During 7 to 28 d and 8 to 12 wk of age, the combination of Thr, Ile, Val, Arg, and Trp to 100% of NRC reversed the BW depression; here only Thr, Ile, and Val were essential components of the response. The BW depression during 16 to 20 wk was reversed by the combination of Thr, Ile, Val, and Trp. For turkeys fed 60% of CP, BW gain was severely depressed. The combination of Thr, Ile, Val, Trp, and Arg resulted in nearly complete BW recovery during each age. PMID- 11020076 TI - Hydrolysis and absorption in the small intestines of posthatch chicks. AB - In the immediate posthatch period, chicks must transfer from metabolic dependence on yolk to utilization of exogenous feed. This study describes changes in intestinal luminal pancreatic enzyme activity and mucosal uptake posthatch as influenced by feed and Na intake. Chicks with access to feed increased in BW and small intestinal weight in the 48-h posthatch, whereas chicks without access to feed decreased in BW; however, small intestinal weight increased during this period. Chicks ingesting feed showed increases in total intestinal trypsin, amylase and lipase activities that were correlated with intestinal weights and BW. Chicks without access to feed showed little change in trypsin and amylase activities, and these increased only after feed consumption. Feeding a low-Na diet did not significantly change the regression coefficient between pancreatic enzyme activity and BW. Mucosal uptake was estimated by measuring Na+,K+ adenosine triphosphatase (ATPase) activity in small intestinal segments. In fed birds this activity increased in relationship to growth, whereas in nonfed birds uptake increased only after access to feed. Low-Na diets allowed only minimal mucosal uptake in all intestinal segments. This study indicates that secretion of trypsin and amylase into the intestine was triggered by feed intake. In addition, Na plays a critical role in intestinal uptake in the immediate posthatch period. PMID- 11020077 TI - Effect of zinc bacitracin and salinomycin on intestinal microflora and performance of broilers. AB - A feeding experiment was carried out over 42 d with four groups of broiler chickens fed experimental diets formulated to provide no supplementation, 20 mg zinc bacitracin, 60 mg salinomycin, or both feed additives in combination. During the fifth week of the experiment, four chickens from each pen were killed, and the contents of gizzard, duodenum, jejunum, ileum, ceca, and rectum were separately collected and pooled. In all intestinal segments, the pH and the concentration of lactic acid were measured, and the numbers of anaerobic bacteria, coliforms, lactic acid bacteria, lactobacilli, enterococci, and Clostridium perfringens were counted. In homogenates of pancreas obtained from four animals, the activities of amylase, lipase, trypsin, and chymotrypsin were measured. A significant growth-promoting effect was observed in the group receiving zinc bacitracin in combination with salinomycin. Zinc bacitracin significantly reduced the number of coliform bacteria in the ileum and increased the activities of amylase and lipase in pancreas homogenates. Supplementation with salinomycin and zinc bacitracin, alone or in combination, resulted in significantly lower counts of C. perfringens as well as Lactobacillus salivarius, which was a dominant lactic acid bacterium found in broiler intestinal contents. High numbers of these lactobacilli may play a role in broiler growth depression related to competition in nutrient uptake or impaired fat absorption due to bile acid deconjugation. PMID- 11020078 TI - Effect of the inclusion time of dietary saturated and unsaturated fats before slaughter on the accumulation and composition of abdominal fat in female broiler chickens. AB - The aim of this experiment was to assess the effects of four different feeding programs designed to include tallow, a saturated fat at 0, 8, 12, and 28 d prior to slaughter on female broiler performance and the deposition, fatty acid profile, and melting point of abdominal fat. The following treatment groups were established according to dietary inclusion--from 21 to 49 d of age--of: sunflower oil (SUN), sunflower oil followed by tallow during the last 8 d (SUN + 8TALL), sunflower oil followed by tallow during the last 12 d (SUN + 12TALL), and tallow (TALL). The diets were designed to be isoenergetic and isonitrogenous. Abdominal fat deposition increased linearly with increasing number of days in which birds were fed the tallow-enriched diet. However, linear and quadratic response patterns were found between days before slaughter in which the birds were fed the tallow-enriched diet and abdominal fat melting points. This result suggested an exponential response in which 85% of the maximum level was already attained when the dietary fat type changed from an unsaturated to a saturated condition during the last 8 d of the feeding period. The use of an unsaturated fat source during the first stages of growth, and the substitution of a saturated fat for a few days before slaughter, may offer the advantage of lower abdominal fat deposition and an acceptable fat fluidity compared with the use of a saturated fat source during the whole growing and finishing period. PMID- 11020079 TI - Utilization of soybean oil and tallow in young turkeys compared with young chickens. AB - The experiment was conducted to determine the apparent metabolizable energy and apparent fatty acid digestibility of tallow and soybean oil (8% of the basal diet) in young (1 and 3 wk of age) turkeys compared with young chickens. At 1 wk of age, turkeys used fats, particularly saturated fats, more efficiently than young chickens (i.e., total fatty acid digestibility was 96.5 vs. 86.4% for soybean oil and 75.0 vs. 49.1% for tallow). This difference between the two species disappeared at 3 wk of age, when there was an increase (95.7 vs. 99.3% for soybean oil and 64.0 vs. 69.7% for tallow) in fat utilization in young chickens, whereas in turkeys it remained constant (96.5 vs. 99.3% for soybean oil and 75 vs. 69.3% for tallow). This result suggests a greater and earlier maturity of the digestive system for fat utilization in turkeys than in young chickens. This efficient utilization of saturated fatty acids in turkeys seemed to depend more on the fat origin than on bird age, because it remained constant with age (91.4 vs. 96%) when saturated fatty acids were provided by soybean oil, and decreased (particularly for C18:0: 5.08 vs. 35.8%) when saturated fatty acids were provided by tallow. In turkeys, fluctuations in secretion of bile salts or in lipase activity during the trial period may have caused the lower stearic acid digestibility at 3 wk of age. The present study revealed a difference in fat utilization between turkeys and young chickens. For higher validity of the digestibility of fats, it would be preferable to use turkeys to derive metabolizable energy and fatty acid digestibility of fat values in formulating turkey diets. PMID- 11020080 TI - Histological alterations in the intestinal epithelium caused by the inclusion of full-fat sunflower kernels in broiler chicken diets. AB - Changes in small intestinal morphology (jejunum) were examined at 28 d of age in chicks fed with full-fat sunflower kernels (FFSK)-based diets. Jejunal mucosa of chicks (six chicks per treatment) were embedded in Epon-812 for examination by a conventional electron microscope procedure. A portion of the tissues was also sectioned and embedded in paraffin for examination by light microscopy. Brush border, goblet cells, and intraepithelial lymphocytes, on the one hand, and fibroblast and mononuclear cells (lymphocytes, plasma cells, macrophages) were observed in the enterocytes and lamina propria, respectively. The results showed that the birds fed 150 g kg(-1) of FFSK showed a shortening and thickening of the villi, hyperplasia and vacuolar degeneration of enterocytes, and hypertrophy and hyperplasia of goblet cells. Likewise, an increment of intraepithelial lymphoid cells and hypercellularity of the lamina propria was observed. In addition, electron microscopy showed large vacuoles in the enterocytes, which could be dilations of agranular and granular endoplasmic reticulum or Golgi. There were many dark granules within the vacuoles that could be triglyceride-rich lipoproteins (portomicrons). These lesions could have been due to the presence of chlorogenic acid or to the greater concentration of oil in the FFSK diet. Our observations demonstrated that addition of 150 g kg(-1) FFSK to broiler chicken diets caused alterations in jejunal mucosa that could explain the decrease in fat digestibility observed in a previous experiment in which we incorporated FFSK into broiler diets. PMID- 11020081 TI - Citric acid does not improve phosphorus utilization in laying hens fed a corn soybean meal diet. AB - Data previously obtained from our laboratory have indicated that citric acid was very effective at improving phytate-P utilization in chicks fed a P-deficient corn-soybean meal (SBM) diet. The objective of the current study was to determine if citric acid would have similar effects in a commercial strain of Single Comb White Leghorn laying hens (n = 432) fed a corn-SBM diet containing 0.10% available P (AP). Dietary treatments were a corn-SBM basal diet (0.10% AP, 3.8% Ca, and 17% CP) supplemented with 0, 1, 2, 3, or 4% citric acid and a positive control diet containing 0.45% AP. Each of the six dietary treatments were fed to six replicate groups of 12 hens from 22 to 40 wk of age. No significant differences in performance were observed among treatments during the first 4 wk of the experiment. Hen-day egg production, feed consumption, feed efficiency, and egg yield were subsequently depressed in hens fed 0, 1, 2, 3, or 4% citric acid compared with hens fed 0.45% AP. The results of this study indicate that citric acid does not improve the utilization of dietary P in laying hens fed a corn-SBM diet containing 3.8% Ca. PMID- 11020082 TI - Relationship between mechanical properties and pentosidine in tendon: effects of age, diet restriction, and aminoguanidine in broiler breeder hens. AB - Nonenzymatic glycosylation contributes to the formation of crosslinks, which leads to the structural and functional deterioration of tissue protein. The accumulation of these crosslinks in tissue proteins has been implicated in the alteration of biomechanical properties of connective tissues. The objective of this study was to determine whether tendon breaking time (TBT) and tendon breaking strength (TBS) of the flexor perforans et perforatus digiti iii tendon were related to concentrations of pentosidine in tendons (Pt) of broiler breeder hens from 8 to 125 wk of age. In addition, effects of diet restriction (DR) and a crosslinking inhibitor, aminoguanidine (AG) on Pt, TBS, and TBT were determined. Female chicks (n = 450) were randomly assigned to four treatment groups immediately after hatch: ad libitum-fed (AL); diet-restricted (DR; 60% of AL); and AL and DR groups supplemented with 1.35 mg/kg BW per day AG in the feed (AL+AG and DR+AG, respectively). In AL hens, Pt increased with increasing age (P < or = 0.0001). Concurrently, an age-related parallel increase was found for TBS (P < or = 0.0001) and TBT (P < or = 0.0001). Rate of Pt accumulation was lower in DR (P < or = 0.001), TBS (P < or = 0.01), and TBT (P < or = 0.02) hens compared with AL hens. Concentration of Pt in the AL + AG group was lower (P < or = 0.0002) than in the AL group; TBS and TBT (P < or = 0.01) followed a similar pattern. Supplementation of DR with AG did not affect Pt, TBS, or TBT. The age related increase in Pt and loss of elasticity in the tendon was retarded by diet restriction and AG. PMID- 11020083 TI - Effect of in ovo administration of insulin-like growth factor-I on composition and mechanical properties of chicken bone. AB - The influence of in ovo administration of insulin-like growth factor-I (IGF-I) on long bone growth (tibiae and femora) of 42-d-old broiler chickens was investigated. Eggs were divided into three groups: uninjected control, vehicle injected control, and recombinant human (rh) IGF-I. Eggs were injected once with 100 microL vehicle (10 mM acetic acid and 0.1% BSA) per embryo or vehicle containing 100 ng rh IGF-I/100 microL per embryo (n = 555 eggs total) on Days 1, 2, 3, or 4 of embryonic development. Males had greater bone length and moment of inertia than did females for the tibia and the femur (P < or = 0.01 for all). Although fracture load was significantly affected by gender (P < or = 0.02 and P < or = 0.006 for the femur and tibia, respectively), there was no treatment effect on these variables. However, when the fracture load was normalized with body weight of the animal, treatment and gender effects were found for femora (P < or = 0.04). Hydroxyproline concentrations of bones from male broilers were increased by the treatment (P < or = 0.02), whereas it had no effect on female broilers. There was no treatment effect on ash content, stiffness, yield load, yield deflection, and ultimate deflection and elastic, plastic, and total work for the femur or the tibia. We suggest that the effect of in ovo administration of IGF-I on bone mechanical properties was site-specific, and treated femora tended to have a lower fracture load relative to increased body weight. PMID- 11020084 TI - An evaluation of the presence of pathogens on broilers raised on poultry litter treatment-treated litter. AB - Two trials were conducted to evaluate the presence of salmonella, campylobacter, and generic Escherichia coli on broilers raised on Poultry Litter Treatment (PLT) enhanced litter in comparison with those raised on untreated litter. Two Company A farms included three houses on each farm as the treated group and three houses per farm as controls. Two complete growouts were evaluated on each farm. The Company B study included 10 farms with two paired houses per farm, one house as the treated group and one house as the control. One growout was evaluated per farm. The pathogen sampling consisted of litter sampling and whole bird rinses on the farm and in the processing plant. Litter pH, ammonia concentration, total litter bacteria, temperatures, and humidity were also recorded. The study with Company A resulted in lower mean levels of pH, ammonia concentration, total litter bacteria, litter E. coli, and bird rinse counts for salmonella and E. coli in houses treated with PLT. The results for Company B closely resembled those for Company A, but also included campylobacter data, which showed no difference between treated and control groups. The data indicate that PLT may be a beneficial component for on-farm pathogen reduction. PMID- 11020085 TI - Adhesion of rice flour-based batter to chicken drumsticks evaluated by laser scanning confocal microscopy and texture analysis. AB - The convenience and appeal of battered or breaded products have resulted in a sales increase of 100% since 1980. Because of the rapid growth of the Asian American population and increasing consumption of rice and rice products, rice flour is a logical alternative for wheat flour in traditional batter formulation. The effects of ingredients used in rice flour-based batters on adhesion characteristic for deep-fat fried chicken drumsticks were studied by laser scanning confocal microscopy (LSCM) and texture analysis. Raw chicken drumsticks were predusted with egg albumin powder before dipping into batters prepared from combinations of rice flour, yellow corn flour, oxidized cornstarch, methylcellulose, or xanthan gum. The drumsticks were fried at 175+/-5 C until the internal temperature reached at least 71 C. For LSCM, samples were fixed overnight and were sectioned by vibratome (200 microm) before viewing. Batter adhesion was determined using an attachment specifically designed for chicken drumsticks. Microstructural analysis showed that batter formulated with a 50:50 mixture of rice and corn flours adhered better to drumsticks than batter with other rice flour ratios. Xanthan gum (0.2%) or methylcellulose (0.3%) alone had poor adhesion to chicken skin. However, when combined with other ingredients, xanthan gum increased the amount of batter pick-up before frying by increasing viscosity. Egg albumin significantly facilitated batter adhesion. The results from texture analysis supported the microstructural studies. As rice flour ratio increased from 50 to 70%, the binding force decreased. Rice flour showed potential as an alternative to wheat flour for batter formulas when the appropriate levels of oxidized starch, xanthan gum, and methylcellulose were included in the formulation. PMID- 11020086 TI - The influence of postmortem electrical stimulation on rigor mortis development, calpastatin activity, and tenderness in broiler and duck pectoralis. AB - This study was conducted to evaluate the effects of electrical stimulation (ES) on rigor mortis development, calpastatin activity, and tenderness in anatomically similar avian muscles composed primarily of either red or white muscle fibers. A total of 72 broilers and 72 White Pekin ducks were either treated with postmortem (PM) ES (450 mA) at the neck in a 1% NaCl solution for 2 s on and 1 s off for a total of 15 s or were used as nonstimulated controls. Both pectoralis muscles were harvested from the carcasses after 0.25, 1.25, and 24 h PM and analyzed for pH, inosine:adenosine ratio (R-value), sarcomere length, gravimetric fragmentation index, calpastatin activity, shear value, and cook loss. All data were analyzed within species for the effects of ES. Electrically stimulated ducks had a lower muscle pH at 0.25 and 1.25 h PM and higher R-values at 0.25 h PM compared with controls. Electrically stimulated broilers had a lower muscle pH at 1.25 h and higher R-values at 0.25 and 1.25 h PM compared with controls. Muscles of electrically stimulated broilers exhibited increased myofibrillar fragmentation at 0.25 and 1.25 h PM, whereas there was no such difference over PM time in the duck muscle. Electrical stimulation did not affect calpastatin activity in either broilers or ducks; however, the calpastatin activity of the broilers did decrease over the aging time period, whereas that of the ducks did not. Electrical stimulation decreased shear values in broilers at 1.25 h PM compared with controls; however, there was no difference in shear values of duck muscle due to ES at any sampling time. Cook loss was lower for electrically stimulated broilers at 0.25 and 1.25 h PM compared with the controls, but had no effect in the ducks. These results suggest that the red fibers of the duck pectoralis have less potential for rigor mortis acceleration and tenderization due to ES than do the white fibers of the broiler pectoralis. PMID- 11020088 TI - Depression: a symptom of coronary artery disease or a pathogenetic factor? PMID- 11020087 TI - Inflammation, depressive symptomtology, and coronary artery disease. AB - OBJECTIVE: Many patients feel exhausted or depressed before the onset of an acute coronary event, but little is known about the origin of these feelings. We tested the hypothesis that the depressive symptomatology is associated with a reactivation of latent viruses and inflammation of a coronary vessel. METHODS: A blood sample was drawn and a biopsy sample was obtained from the coronary lesion of 15 exhausted and 15 nonexhausted patients treated with directional coronary angioplasty because of severe angina. Blood samples were analyzed to measure antibody titers against Chlamydia pneumoniae, cytomegalovirus, and the cytokines interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha. The biopsy sample was analyzed for the presence of IL-1beta and TNF-alpha. RESULTS: Exhausted/depressed patients had higher antibody titers against cytomegalovirus, higher levels of C. pneumoniae immunoglobulin G, and higher levels of IL-1beta and TNF-alpha. No associations between the mental state of a patient and cytokine mRNA in the biopsy sample were found. CONCLUSIONS: The findings indicate that the mental state of angioplasty patients is positively associated with serological markers of inflammation. It remains to be seen whether the inflammation causes feelings of exhaustion, whether exhaustion and depression set the stage for inflammation, or whether existing feelings of exhaustion are amplified by the inflammation. PMID- 11020089 TI - Are vacations good for your health? The 9-year mortality experience after the multiple risk factor intervention trial. AB - OBJECTIVE: The objective of this study was to determine the risk for various causes of posttrial death associated with vacation frequency during the Multiple Risk Factor Intervention Trial (MRFIT). METHODS: Middle-aged men at high risk for coronary heart disease (CHD) were recruited for the MRFIT. As part of the questionnaires administered during the first five annual visits, men were asked whether they had had a vacation during the past year. For trial survivors (N = 12,338), the frequency of these annual vacations during the trial were used in a prospective analysis of posttrial all-cause and cause-specific mortality during the 9-year follow-up period. RESULTS: The relative risk (RR) associated with more annual vacations during the trial was 0.83 (95% confidence interval [CI], 0.71 0.97) for all-cause mortality during the 9-year follow-up period. For cause of death, the RRs were 0.71 (95% CI, 0.58-0.89) and 0.98 (95% CI, 0.78-1.23) for cardiovascular and noncardiovascular causes, respectively. The RR was 0.68 (95% CI, 0.53-0.88) for CHD (including acute myocardial infarction). These associations remained when statistical adjustments were made for possible confounding variables, including baseline characteristics (eg, income), MRFIT group assignment, and occurrence of a nonfatal cardiovascular event during the trial. CONCLUSIONS: The frequency of annual vacations by middle-aged men at high risk for CHD is associated with a reduced risk of all-cause mortality and, more specifically, mortality attributed to CHD. Vacationing may be good for your health. PMID- 11020090 TI - A randomized, wait-list controlled clinical trial: the effect of a mindfulness meditation-based stress reduction program on mood and symptoms of stress in cancer outpatients. AB - OBJECTIVE: The objective of this study was to assess the effects of participation in a mindfulness meditation-based stress reduction program on mood disturbance and symptoms of stress in cancer outpatients. METHODS: A randomized, wait-list controlled design was used. A convenience sample of eligible cancer patients enrolled after giving informed consent and were randomly assigned to either an immediate treatment condition or a wait-list control condition. Patients completed the Profile of Mood States and the Symptoms of Stress Inventory both before and after the intervention. The intervention consisted of a weekly meditation group lasting 1.5 hours for 7 weeks plus home meditation practice. RESULTS: Ninety patients (mean age, 51 years) completed the study. The group was heterogeneous in type and stage of cancer. Patients' mean preintervention scores on dependent measures were equivalent between groups. After the intervention, patients in the treatment group had significantly lower scores on Total Mood Disturbance and subscales of Depression, Anxiety, Anger, and Confusion and more Vigor than control subjects. The treatment group also had fewer overall Symptoms of Stress; fewer Cardiopulmonary and Gastrointestinal symptoms; less Emotional Irritability, Depression, and Cognitive Disorganization; and fewer Habitual Patterns of stress. Overall reduction in Total Mood Disturbance was 65%, with a 31% reduction in Symptoms of Stress. CONCLUSIONS: This program was effective in decreasing mood disturbance and stress symptoms in both male and female patients with a wide variety of cancer diagnoses, stages of illness, and ages. cancer, stress, mood, intervention, mindfulness. PMID- 11020091 TI - Stress and body shape: stress-induced cortisol secretion is consistently greater among women with central fat. AB - OBJECTIVE: Excessive central fat puts one at greater risk of disease. In animal studies, stress-induced cortisol secretion has been shown to increase central fat. The objective of this study was to assess whether women with central fat distribution (as indicated by a high waist-to-hip ratio [WHR]), across a range of body mass indexes, display consistently heightened cortisol reactivity to repeated laboratory stressors. METHODS: Fifty-nine healthy premenopausal women, 30 with a high WHR and 29 with a low WHR, were exposed to consecutive laboratory sessions over 4 days (three stress sessions and one rest session). During these sessions, cortisol and psychological responses were assessed. RESULTS: Women with a high WHR evaluated the laboratory challenges as more threatening, performed more poorly on them, and reported more chronic stress. These women secreted significantly more cortisol during the first stress session than women with a low WHR. Furthermore, lean women with a high WHR lacked habituation to stress in that they continued to secrete significantly more cortisol in response to now familiar challenges (days 2 and 3) than lean women with a low WHR. CONCLUSIONS: Central fat distribution is related to greater psychological vulnerability to stress and cortisol reactivity. This may be especially true among lean women, who did not habituate to repeated stress. The current cross-sectional findings support the hypothesis that stress-induced cortisol secretion may contribute to central fat and demonstrate a link between psychological stress and risk for disease. PMID- 11020092 TI - Exercise treatment for major depression: maintenance of therapeutic benefit at 10 months. AB - OBJECTIVE: The purpose of this study was to assess the status of 156 adult volunteers with major depressive disorder (MDD) 6 months after completion of a study in which they were randomly assigned to a 4-month course of aerobic exercise, sertraline therapy, or a combination of exercise and sertraline. METHODS: The presence and severity of depression were assessed by clinical interview using the Diagnostic Interview Schedule and the Hamilton Rating Scale for Depression (HRSD) and by self-report using the Beck Depression Inventory. Assessments were performed at baseline, after 4 months of treatment, and 6 months after treatment was concluded (ie, after 10 months). RESULTS: After 4 months patients in all three groups exhibited significant improvement; the proportion of remitted participants (ie, those who no longer met diagnostic criteria for MDD and had an HRSD score <8) was comparable across the three treatment conditions. After 10 months, however, remitted subjects in the exercise group had significantly lower relapse rates (p = .01) than subjects in the medication group. Exercising on one's own during the follow-up period was associated with a reduced probability of depression diagnosis at the end of that period (odds ratio = 0.49, p = .0009). CONCLUSIONS: Among individuals with MDD, exercise therapy is feasible and is associated with significant therapeutic benefit, especially if exercise is continued over time. PMID- 11020093 TI - Change in heart rate and heart rate variability during treatment for depression in patients with coronary heart disease. AB - OBJECTIVE: Major depression is a common problem in patients with coronary heart disease (CHD) and is associated with an increased risk for cardiac morbidity and mortality. It is not known whether treating depression will improve medical prognosis in patients with CHD. Depression is also associated with elevated heart rate and reduced heart rate variability (HRV), which are known risk factors for cardiac morbidity and mortality that may explain the increased risk associated with depression. The purpose of this study was to determine whether treatment for depression with cognitive behavior therapy (CBT) is associated with decreased heart rate or increased HRV. METHODS: Thirty depressed patients with stable CHD, classified as either mildly or moderately to severely depressed, received up to 16 sessions of CBT. The 24-hour heart rate and HRV were measured in these patients and in 22 medically comparable nondepressed controls before and after treatment of the depressed patients. RESULTS: Average heart rate and daytime rMSSD (reflecting mostly parasympathetic activity) improved significantly in the severely depressed patients, but remained unchanged in the mildly depressed and the control patients. However, only rMSSD improved to a level comparable to the control patients. None of the remaining indices of HRV showed improvement. CONCLUSIONS: The results suggest that treating depression with CBT may reduce heart rate and increase short-term HRV. Thus, CBT may have a beneficial effect on a risk factor for mortality in depressed patients with coronary heart disease. A randomized, controlled study is needed to confirm these findings. PMID- 11020094 TI - Cardiovascular recovery from acute laboratory stress: reliability and concurrent validity. AB - OBJECTIVE: We assessed the value of laboratory measures of cardiovascular recovery across four criteria: reliability across multiple tasks, reliability across a 3-year time interval, ability to predict daily ambulatory blood pressure, and interrelationships with coronary risk factors and psychosocial variables. METHODS: Three hundred twenty-nine healthy adults (mean age = 27.1 years) completed a two-part protocol consisting of 1 day of laboratory testing and 1 day of ambulatory monitoring. The laboratory protocol included a 15-minute baseline assessment followed by three 5-minute laboratory challenges (mental arithmetic, speech, and handgrip). Five-minute recovery periods followed each exercise. One hundred twenty-five participants returned after 3 years to repeat the protocol. RESULTS: When aggregated across tasks, cardiovascular recovery showed acceptable levels of internal consistency (alpha values = 0.7) and proved relatively stable across time (r values = 0.22-0.35). Recovery values statistically improved the prediction of daily ambulatory readings above baseline and stress reactivity laboratory values (p values < .001) but were largely unrelated to coronary risk factors or psychosocial measures. CONCLUSION: These results suggest that cardiovascular recovery from acute laboratory stress can be treated as a stable individual difference variable that can -improve standard laboratory-based predictor models of ambulatory readings. PMID- 11020095 TI - Chronic fatigue syndrome, fibromyalgia, and multiple chemical sensitivities in a community-based sample of persons with chronic fatigue syndrome-like symptoms. AB - OBJECTIVE: The aim of this study was to determine illness comorbidity rates for individuals with chronic fatigue syndrome (CFS), fibromyalgia (FM), and multiple chemical sensitivities (MCS). An additional objective was to identify characteristics related to the severity of fatigue, disability, and psychiatric comorbidity in each of these illness groups. METHODS: A random sample of 18,675 residents in Chicago, Illinois, was first interviewed by telephone. A control group and a group of individuals with chronic fatigue accompanied by at least four minor symptoms associated with CFS received medical and psychiatric examinations. RESULTS: Of the 32 individuals with CFS, 40.6% met criteria for MCS and 15.6% met criteria for FM. Individuals with MCS or more than one diagnosis reported more physical fatigue than those with no diagnosis. Individuals with more than one diagnosis also reported greater mental fatigue and were less likely to be working than those with no diagnosis. Individuals with CFS, MCS, FM, or more than one diagnosis reported greater disability than those with no diagnosis. CONCLUSIONS: Rates of coexisting disorders were lower than those reported in prior studies. Discrepancies may be in part attributable to differences in sampling procedures. People with CFS, MCS, or FM endure significant disability in terms of physical, occupational, and social functioning, and those with more than one of these diagnoses also report greater severity of physical and mental fatigue. The findings illustrate differences among the illness groups in the range of functional impairment experienced. PMID- 11020096 TI - Acute psychological stress and exercise and changes in peripheral leukocyte adhesion molecule expression and density. AB - OBJECTIVE: This study examined the effects of acute psychological stress and exhaustive exercise on the expression and density of adhesion molecules (L selectin, lymphocyte function antigen-1 [LFA-1], and intracellular adhesion molecule-1 [ICAM-1]) on monocytes, granulocytes, and lymphocytes. METHODS: Forty five healthy volunteers performed a 15-minute public speaking task and a 15- to 18-minute bicycle ergometer challenge. RESULTS: In general, both the exercise and speaking tasks led to increases in the number of circulating leukocytes and lymphocyte subsets. The density of L-selectin (CD62L) on mixed lymphocytes and T lymphocytes was decreased in response to exercise (p values < .001). Both stressors led to an increased density of LFA-1 (CD11a) on mixed lymphocytes (p values < .01), whereas CD11a density on monocytes and granulocytes remained unchanged. ICAM-1 (CD54) density was unaffected, but the number of lymphocytes, monocytes, and granulocytes expressing CD54 increased in the circulation on both stressors. CONCLUSIONS: The data indicate that both psychological stress and exercise have significant effects on cellular expression of adhesion molecules on circulating leukocytes. Given the crucial role that adhesion molecules on circulating cells play in inflammation and disease, these findings may have clinical relevance in sympathetic nervous system-induced immune activation. PMID- 11020097 TI - Pavlovian conditioning of taste aversion using a motion sickness paradigm. AB - OBJECTIVE: Pavlovian conditioning of taste aversion has rarely been investigated in healthy humans using motion sickness as the unconditioned stimulus (US). METHODS: Ninety subjects were pretested for susceptibility to illusory motion (vection) in a rotating drum. Thirty-two subjects susceptible to pseudomotion were assigned randomly to two groups and received either water 1 hour before rotation and a novel taste (elderberry juice, conditioned stimulus, [CS]) immediately before rotation in a rotating chair (conditioning group), or the sequence of water and juice was reversed (control group). During the test session 1 week later, all subjects were exposed to water 1 hour before and juice immediately before rotation. The amount of liquids ingested, nausea ratings, rotation tolerance, and blood levels of hormones (ACTH, ADH, PP) were evaluated. RESULTS: Subjects in the conditioning group developed taste aversion toward the novel taste, but not subjects in the control group. Postrotation nausea rating was affected marginally by conditioning, but rotation tolerance was not changed by conditioning. ACTH and ADH but not PP levels increased with rotation, but were unaffected by conditioning. CONCLUSIONS: Pavlovian conditioning of behavioral, but not of endocrine, indicators was effective in susceptible subjects using a rotating chair as US and a single CS-US pairing. PMID- 11020098 TI - Do panic symptom profiles influence response to a hypoxic challenge in patients with panic disorder? A preliminary report. AB - OBJECTIVE: This study examined how panic symptom profiles affect response to a hypoxic laboratory challenge in patients with panic disorder. METHODS: Seven patients whose naturally occurring panic attacks were characterized by prominent respiratory symptoms (Resp subgroup) were compared and contrasted with seven patients who did not report respiratory symptoms during panic attacks (NonResp subgroup). All were administered a novel 12% O2 challenge and assessed with measures of tidal volume, respiratory rate, end-tidal CO2, anxiety, and panic symptoms. RESULTS: Although the Resp and NonResp subgroups showed equivalent increases in anxiety and panic symptoms, the Resp subgroup showed greater fluctuation in tidal volume during and after the challenge as well as overall lower levels of end-tidal CO2. CONCLUSIONS: Our results suggest the importance of panic symptom profiles in determining respiratory responses to a hypoxic challenge in patients with panic disorder. These findings are discussed in light of current theories of panic disorder, with particular attention to respiratory disturbances in this disorder. PMID- 11020099 TI - Anger, harassment, and cardiovascular reactivity among Chinese and Indian men in Singapore. AB - OBJECTIVE: This experiment examined psychological and cardiovascular responses to experimental harassment among Chinese and Indian men in Singapore who differed in levels of dispositional anger. METHODS: Eighty-four Chinese and Indian men participated in a laboratory experiment on cardiovascular reactivity in which mood was rated and heart rate and blood pressure were measured during computer tasks in which they were either harassed or allowed to complete the tasks without interruption. RESULTS: Comparison of systolic reactivity to harassment and nonharassment indicated, as expected, that reactivity was greater after harassment. Furthermore, a significant race by dispositional anger by harassment effect was obtained for systolic reactivity that indicated different patterns of reactivity for Chinese and Indian participants. In the absence of harassment, Chinese participants showed low systolic reactivity regardless of their level of dispositional anger, whereas systolic reactivity increased as a function of dispositional anger when they were harassed. For Indians, however, systolic reactivity was a positive function of dispositional anger both when they were harassed and not harassed. CONCLUSIONS: These results suggest stronger cardiovascular reactivity to stress among Indian than among Chinese men. This seems to be particularly true for Indians high in dispositional anger. PMID- 11020100 TI - Psychological and perceptual factors associated with arrhythmias and benign palpitations. AB - OBJECTIVE: Little is known about how patients who seek medical help for benign palpitations can be distinguished from those with clinically significant arrhythmias. This study tested whether patients with arrhythmia can be distinguished from those who are aware of sinus rhythm or extrasystoles on the basis of sex, prevalence of anxiety disorders, and heartbeat perception. METHODS: A consecutive sample of patients referred to a cardiology clinic participated in the study. Patients were diagnosed as having either arrhythmia (N = 62), extrasystoles (N = 75), or awareness of sinus rhythm (N = 47). They were assessed with use of the anxiety disorders and hypochondriasis modules of the Structured Clinical Interview for DSM-IV. Both patients and control subjects (N = 35) answered questionnaires measuring anxiety, fear of bodily sensations, and depression and underwent a heartbeat perception test. The present report focuses on patients who had palpitations but no comorbid cardiovascular disease. RESULTS: Patients with awareness of sinus rhythm could be distinguished from those with arrhythmia by several variables: female sex, higher prevalence of panic disorder, poor performance on the heartbeat perception test, report of palpitations when doing the test, higher heart rates, lower levels of physical activity, and (as trends) a greater prevalence of panic attacks, fear of bodily sensations, and depression. In contrast, patients with arrhythmias rarely reported palpitations when doing the test but were more likely to perceive their heartbeats accurately than patients with sinus rhythm and control subjects. Performance on the heartbeat perception test was intermediate in patients with extrasystoles; these patients also had an intermediate prevalence of panic disorder and intermediate depression scores. CONCLUSIONS: Measures of panic disorder and a simple heartbeat perception test could complement medical assessment in the diagnosis of patients who seek medical help for palpitations. The results also have implications for the treatment of patients with benign palpitations. PMID- 11020101 TI - Personality and social predictors of atherosclerotic progression: Edinburgh Artery Study. AB - OBJECTIVE: The aim of this study was to determine whether personality traits and social factors predict the progression of peripheral atherosclerosis. Progression was assessed using the objective, noninvasive ankle brachial pressure index (ABPI). METHODS: In the Edinburgh Artery Study, 1592 men and women were randomly sampled from the general population, and their ABPI was measured at baseline and at the end of a 5-year follow-up period. A low ABPI suggests the presence of peripheral arterial disease. The revised Bedford-Foulds Personality Deviance Scale was administered at baseline to assess submissiveness and hostility. Data on other baseline risk factors, including physiological and social factors, were also collected. RESULTS: Change in ABPI over 5 years was negatively correlated with age in both men and women (men, r = -0.10; women, r = -0.25). In multiple linear regression models, smoking, alcohol consumption, and submissiveness together accounted for 2% of the variance in ABPI change in men; in women, only age was related to change, accounting for 6% of the variance. Well-fitting structural equation models in both sexes showed that age influenced baseline ABPI and change in ABPI; that smoking and social deprivation directly affected baseline ABPI; and that the effect of hostility, and some of the effect of social deprivation, was mediated by smoking. CONCLUSIONS: Social and personality factors were associated directly with baseline ABPI levels and indirectly with progression of atherosclerosis. Structural equation models revealed that associations among personality, social factors, and atherosclerotic progression were complex, involving mediation through other variables. PMID- 11020102 TI - Maternal social support predicts birth weight and fetal growth in human pregnancy. AB - OBJECTIVE: Low birth weight is a primary cause of infant mortality and morbidity. Results of previous studies suggest that social support may be related to higher birth weight through fetal growth processes, although the findings have been inconsistent. The purpose of this investigation was to test a model of the association between a latent prenatal social support factor and fetal growth while taking into account relations between sociodemographic and obstetric risk factors and birth weight. METHOD: A prospective study was conducted among 247 women with a singleton, intrauterine pregnancy receiving care in two university affiliated prenatal clinics. Measures of support included support from family, support from the baby's father, and general functional support. Sociodemographic characteristics were also assessed. Birth outcome and obstetric risk information were abstracted from patients' medical charts after delivery. RESULTS: Structural equation modeling analyses showed that a latent social support factor significantly predicted fetal growth (birth weight adjusted for length of gestation) with infant sex, obstetric risk, and ethnicity in the model. Marital status and education were indirectly related to fetal growth through social support. The final model with social support and other variables accounted for 31% of the variance in fetal growth. CONCLUSIONS: These findings suggest that prenatal social support is associated with infant birth weight through processes involving fetal growth rather than those involving timing of delivery. Biological and behavioral factors may contribute to the association between support and fetal growth, although these mechanisms need to be further explored. These results pave the way for additional research on fetal growth mechanisms and provide a basis for support intervention research. PMID- 11020103 TI - Psychological differences between veterans with and without Gulf War unexplained symptoms. Portland Environmental Hazards Research Center. AB - OBJECTIVE: The objective of this study was to assess measures of psychological and neurobehavioral functioning to determine their association with unexplained symptoms in Gulf War veterans. METHODS: An epidemiological survey focusing on exposures and symptoms was mailed to a random sample of Gulf War veterans from Oregon and southwestern Washington. Volunteers were recruited from survey respondents who agreed to undergo a thorough medical examination and psychological and neurobehavioral assessment. Persistent symptoms with no medical explanation associated with service in the Persian Gulf (eg, fatigue, muscle pain, and memory deficits) that began during or after the war qualified respondents as cases. The 241 veterans with unexplained symptoms were classified as case subjects, and the 113 veterans without symptoms were classified as control subjects. All veterans completed a battery of computerized assessment tests consisting of 12 psychosocial and 6 neurobehavioral tests. Differences between case and control subjects on neurobehavioral and psychological variables were assessed with univariate and multivariate statistical comparisons. RESULTS: Case subjects differed substantially and consistently from control subjects on diverse psychological tests in the direction of increased distress and psychiatric symptoms. Case subjects had small but statistically significant deficits relative to control subjects on some neurobehavioral tests of memory, attention, and response speed. A logistic regression model consisting of four psychological variables but no neurobehavioral variables classified case and control subjects with 86% accuracy. CONCLUSIONS: Our results revealed that Gulf War veterans who report symptoms associated with that conflict differed on multiple psychological measures in the direction of increased distress and performed more poorly on neurobehavioral measures when compared with control subjects who did not report symptoms. This suggests that psychological differences have a prominent role in investigation of possible explanations of Gulf War symptoms. PMID- 11020104 TI - Effect of risperidone on prolactinoma growth in a psychotic woman. AB - OBJECTIVE: The case of a psychotic woman is described in which risperidone use was found to correspond with an increase in the size of a prolactinoma and prevented the return of serum prolactin level to baseline. METHODS: Although the patient had been treated with a high dose of bromocriptine, her prolactin level remained elevated, causing persistent galactorrhea. The patient later was treated with olanzapine and carbamazepine successfully. RESULTS: This case report highlights the role of risperidone on prolactin and discusses alternative methods of treating psychosis when the etiology is unclear, especially in younger patients. CONCLUSIONS: The authors recommend that additional studies regarding the relationship between the growth of prolactinoma and atypical antipsychotics would be worthwhile. PMID- 11020105 TI - Optic neuritis in children. AB - PURPOSE: To describe the clinical characteristics of optic neuritis in children, including final visual acuity and development of multiple sclerosis (MS). METHODS: Charts were reviewed of all patients < 15 years of age who presented with optic neuritis to the Bascom Palmer Eye Institute or the Miami Children's Hospital between 1986 and 1998. RESULTS: Fifteen patients were identified. There was a slight female predilection in the study group (60%), with a mean age of 9.8 years at presentation. A preceding febrile illness within 2 weeks of visual symptoms was reported in 66% of patients. Initial visual acuity ranged from 20/15 to no light perception. Involvement was bilateral in 66% of patients, and disc swelling was present in 64% of involved eyes. Of the patients who underwent magnetic resonance imaging, 33% had focal demyelinating lesions in the brain, and 63% of affected nerves were enlarged or enhanced with gadolinium. Eleven patients were treated with intravenous steroids. Final visual acuity was > or = 20/40 in 58.3% of eyes. Thirty percent of the patients had vision of finger counting or worse. Four (26%) patients developed MS. The mean age of patients with MS was 12 years, compared with 9 years in children who did not develop MS. Patients with unilateral involvement had an excellent visual prognosis (100% > 20/40), but a higher rate of development of MS (75%). Two patients had positive serology for Lyme disease. CONCLUSIONS: Optic neuritis presents differently in children than in adults. Children typically have bilateral involvement with papillitis following an antecedent viral illness. Although visual prognosis is poorer in children than adults, the development of MS is less common in children. Children who present with unilateral involvement have a better visual prognosis; however, they also develop MS at a greater frequency than children with bilateral involvement. Patients who developed MS were, on average, older at presentation with optic neuritis than those who did not develop MS. PMID- 11020106 TI - Ocular fixation to nasal periosteum with a superior oblique tendon in patients with third nerve palsy. AB - PURPOSE: To report a new surgical approach that uses ocular fixation to the nasal periosteum with superior oblique tendon for patients with complete third nerve palsy. METHODS: Prospective study of 15 patients with complete third nerve palsy who underwent surgery using a superior oblique tenectomy and ocular fixation to the nasal periosteum with the superior oblique tendon fragment. RESULTS: Eleven (73%) patients achieved good ocular alignment, 1 (7%) patient had a cosmetically acceptable result, and 3 (20%) patients had a cosmetically unacceptable result. Five (30%) patients had preoperative diplopia; all achieved resolution of their double vision in the primary position of gaze after surgery. Two patients without preoperative diplopia did not achieve good alignment and had diplopia postoperatively. Follow-up ranged from 8-41 months (mean: 19 months). No operative complications occurred. CONCLUSION: Ocular fixation to the nasal periosteum with superior oblique tendon is a safe, effective, and technically undemanding option for the surgical management of patients with complete third nerve palsy. PMID- 11020107 TI - Visual field defects in children with congenital glaucoma. AB - PURPOSE: To determine visual field defects in a cohort of children with congenital glaucoma. METHODS: Monocular visual fields were measured in 24 meridians for targets V4e, I4e, I2e, and I1e, using a Goldmann perimeter in a group of 13 children between the ages of 4 and 14 years with congenital glaucoma and 10 age-matched healthy children. Localized visual field defects (eg, paracentral scotoma, nasal step, and arcuate scotoma) were determined by abnormal findings or shape of the eye in at least one of each of the targets presented. RESULTS: Visual field extent for target 12e was significantly constricted for unilateral and bilateral cases of congenital glaucoma when compared with control eyes. A post-hoc procedure (Tukey Test) showed significant differences between unilateral cases and normal control eyes, and between bilateral cases (best outcome eye) and normal control eyes. Stimuli V4e and 14e results were comparable for patients and normals. Stimulus I1e showed significantly different total extent visual field for bilateral and normal controls. Specific visual field defects were found only in bilateral cases. Paracentral scotoma was found in 1 of 12 eyes with bilateral congenital glaucoma. Nasal steps were found in 6 of 12 eyes with bilateral congenital glaucoma. Arcuate scotoma were found in 4 of 12 eyes with bilateral congenital glaucoma. CONCLUSION: Localized visual fields were found in 37.5% of eyes with congenital glaucoma. Early treatment for congenital glaucoma provided better visual field outcome. PMID- 11020108 TI - Extraocular muscle cysticercosis: a clinical masquerade. AB - PURPOSE: To highlight the different presentations of extraocular muscle cysticercosis, which can result in acquired ocular motility disorders. METHODS: Ten cases of acquired motility disorders examined between January 1998 and January 1999 were diagnosed as extraocular muscle cysticercosis with computed tomography (CT). All patients were treated with corticosteroid therapy and albendazole. Repeat CT scans were performed in 5 patients. RESULTS: The inferior rectus muscle was most commonly affected with double elevator palsy the most common clinical presentation. Response to treatment was satisfactory with complete resolution of the ocular motility disorder in 8 patients. Initiating therapy with corticosteroids prior to the addition of cysticidal drugs prevented severe inflammatory reactions and residual strabismus. CONCLUSION: A high index of suspicion should be entertained for extraocular muscle cysticercosis in every case of recent acquired ocular motility disorder. Response to medical therapy is satisfactory. PMID- 11020109 TI - Disinsertion of the inferior oblique muscle for treatment of superior oblique paresis. AB - PURPOSE: To examine the safety and efficacy of disinsertion of the inferior oblique muscle in the treatment of long-standing ipsilateral superior oblique paresis. METHODS: This retrospective study included 52 patients who underwent disinsertion of the inferior oblique muscle over a 5-year period. Mean age at presentation was 30.8 years (range: 1-70 years). All patients had long-standing unilateral superior oblique paresis and overaction of the ipsilateral inferior oblique muscle. Information recorded included pre- and postoperative vertical deviation, pre- and postoperative Hess chart error scores, reoperation rate, and symptom relief. RESULTS: Following surgery, mean vertical deviation was reduced from 12.9 to 4 prism diopters, while mean Hess chart error score decreased from 596 to 258. This procedure alone resulted in satisfactory symptom relief in 84.6% of patients. The only complication was significant undercorrection requiring further surgery in 13.6% of patients and prisms in 1.8%. When further surgery was needed, the procedure of choice was tucking of the affected superior oblique. Recession of the contralateral inferior rectus was carried out when superior oblique tucking was believed to be unsuitable. CONCLUSIONS: Disinsertion of the inferior oblique muscle is a safe and effective treatment for the majority of long-standing idiopathic unilateral superior oblique pareses. PMID- 11020110 TI - Documenting pediatric lens problems with the MTI polaroid photoscreener. AB - PURPOSE: To evaluate the use of the MTI photoscreener (Medical Technology Inc, Iowa City, Iowa) in assessing and documenting lens abnormalities in the pediatric ophthalmology clinic. METHODS: MTI photoscreener photographs of pediatric patients with lens abnormalities were taken to assess and document lens opacities and subluxation, to assist with explanations to parents, and to monitor progression of abnormalities. Postoperative photographs were taken to assess opacification of the posterior capsule and contact lens fit. RESULTS: Nineteen children with lens abnormalities were examined. Twelve patients underwent surgery and 9 patients had postoperative photoscreener pictures. Illustrative cases are reported. CONCLUSION: The MTI photoscreener is a useful adjunct to clinical examination in the assessment and documentation of pediatric lens abnormalities. PMID- 11020111 TI - Ocular findings in glutaric aciduria type 1. AB - PURPOSE: To determine the nature and course of ocular abnormalities in glutaric aciduria (acidemia) type 1 (GA1). METHODS: Fifteen children with GA1 have been studied in the Republic of Ireland. A retrospective review of the records of the 6 children who died during their illness and prospective clinical examination of 9 survivors were performed. RESULTS: Seven of the 15 children had abnormal eye findings. Ocular complications included intraretinal hemorrhages, cataract, gaze palsy, strabismus, ametropia, and pigmentary retinopathy. CONCLUSION: Ocular involvement is common in glutaric aciduria. Complete ophthalmologic evaluation is recommended in all patients suspected to have this rare disease. Intraretinal hemorrhages due to GA1 could be misinterpreted as resulting from child abuse, and it is important to include this disorder with the differential diagnosis of child abuse. PMID- 11020112 TI - Retrospective comparison of surgical techniques to prevent secondary opacification in pediatric cataracts. AB - PURPOSE: To evaluate the effect of different surgical methods for management of the posterior capsule and anterior vitreous on the rate of posterior capsule opacification in pediatric cataracts. METHODS: Charts of 34 children (47 eyes) aged 40 days to 18 years (mean: 8.5 years) who had primary cataract surgery with or without posterior chamber intraocular lens (IOL) implantation during the past 5 years were reviewed. In 26 eyes, cataracts were managed with a posterior continuous curvilinear capsulorhexis, and in 21 eyes, the posterior capsule was left intact. Follow-up averaged 10 months (range: 6.5 months to 5 years). RESULTS: Visually significant secondary cataract developed in nine eyes with intact posterior capsules, and seven eyes required Nd:YAG laser posterior capsulotomy. The average time for YAG capsulotomy postcataract removal in the second group was 4 months. The visual axis remained clear in all eyes that had posterior continuous curvilinear capsulorhexis with or without posterior chamber IOL. Complications such as fibrinoid membrane, stromal edema, posterior synechiae, updrawn pupil, and transient glaucoma occurred in both groups at a similar rate. CONCLUSION: Primary posterior continuous curvilinear capsulorhexis is an effective method for preventing secondary cataract formation in pediatric cataracts. PMID- 11020113 TI - Isolated granulocytic myelodysplasia in a child with unilateral retinoblastoma. PMID- 11020114 TI - Mobius syndrome: MRI findings in three cases. PMID- 11020115 TI - Palpebral myiasis. PMID- 11020116 TI - Diffuse infiltrating retinoblastoma presenting as a spontaneous hyphema. PMID- 11020117 TI - Intestinal malrotation: another systemic anomaly associated with Peters' syndrome. PMID- 11020118 TI - Preseptal orbital cellulitis following oral trauma. PMID- 11020119 TI - Management of narcolepsy in pregnancy. PMID- 11020120 TI - Platelet supersensitivity to thrombin stimulation in depression: a possible mechanism for the association with cardiovascular mortality. AB - The mortality risk associated with cardiovascular disease is significantly increased in patients with major depression and panic disorder. The mechanism of this phenomenon is unclear. Thrombin is responsible for platelet aggregation and shape change, and it plays a significant role in the development of thromboembolic events. In this study, we examined the platelet second messenger intracellular calcium response to thrombin stimulation in patients with major depression (n = 13), major depression after response to electroconvulsive therapy (ECT; n = 13), subsyndromal depression (n = 16), schizophrenia (n = 15), and control subjects (n = 65). Patients with major depression had significantly higher intracellular calcium responses to thrombin stimulation than control subjects, patients with subsyndromal depression, and patients with schizophrenia (p < 0.05). Electroconvulsive therapy did not significantly change this supersensitivity. This suggests that the platelet response to activation in patients with major depression is supersensitive. This study suggests a possible mechanism for the increased risk of cardiovascular disease that is seen in these two psychiatric disorders. The lack of difference between the control and subsyndromal depression groups appears to validate current diagnostic thresholds in depression. The failure of nonpharmacologic treatment to alter this marker suggests that it may be a trait marker of depression. PMID- 11020121 TI - Mexiletine in the treatment of torticollis and generalized dystonia. AB - Mexiletine is an antiarrhythmic drug that has been reported to exert antidystonic properties. We performed an open-label study to collect further evidence of the antidystonic effect of mexiletine in spasmodic torticollis (ST) and to evaluate its possible use in generalized dystonia. We administered mexiletine to six patients with dystonia (three with generalized dystonia and three with ST) who had failed to respond to previous pharmacotherapy. The drug was started at a dose of 200 mg/d by mouth and increased up to a maximum dose of 800 mg/d. Patients were evaluated at regular intervals over a 6-week period with use of the Fahn & Marsden Dystonia Scale and the Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) and videotaped. At the end of the trial, the videotapes were reviewed and scored by a blind observer. Patients were then followed for at least 1 year and evaluated every 3 months at the dose reached during the study period. No adverse effects were reported in five patients; in one patient, dizziness developed at the dosage of 800 mg/d, requiring a reduction of the dose. At the end of a 6-week period, a significant improvement in the rating scale for dystonia and in videotape ratings was observed after mexiletine treatment (p < 0.01). Our data indicate that mexiletine is a useful drug in dystonia treatment. PMID- 11020122 TI - Decreased homovanilic acid in cerebrospinal fluid correlates with impaired neuropsychologic function in HIV-1-infected patients. AB - To determine whether dopamine metabolism is abnormal in HIV infected patients and whether dopamine metabolism abnormalities are related to specific neuropsychologic characteristics in HIV-infected patients, we measured cerebrospinal fluid (CSF) levels of homovanilic acid (HVA), the primary dopamine metabolite, in 10 HIV-infected patients and compared it to HVA levels in CSF in a group of 13 healthy control subjects. HIV-infected patients were also assessed with a battery of neuropsychologic tests and HVA levels were then correlated with performance on specific neuropsychologic tests. The mean (+/-SD) HVA level in CSF was 100.9 +/- 29.3 nmol/L in the HIV-infected study group and 230.5 +/- 50.0 nmol/L in the non-HIV-infected control group (p < 0.0001). The decrease in concentrations of HVA in CSF correlated with impairment on performance on neuropsychologic testing (Spearman r = 0.67; p = 0.03). When the relationship between HVA levels and specific cognitive domains was evaluated, we observed trends for positive correlation between HVA levels and tests that measure motor speed (r = 0.59; p = 0.074) and those testing attention, concentration, and executive control (r = 0.54; p = 0.108). There was no relationship between performance on memory tests and CSF HVA levels (r = -0.0061; p = 0.987). These results further support the hypothesis that dopaminergic dysfunction plays an important role in the pathogenesis of AIDS dementia complex (ADC) and suggest that specific motor and cognitive abnormalities may be related to depressed dopaminergic activity. This may have important implications for the development of treatments or preventive strategies for ADC. PMID- 11020123 TI - Sustained cabergoline treatment reverses levodopa-induced dyskinesias in parkinsonian monkeys. AB - The pathophysiology of L-Dopa-induced dyskinesias (LID), a common problem after long-term use of L-dopa in the treatment of Parkinson's disease (PD), is not completely understood. Oscillations in L-Dopa concentrations in the brain are believed to be responsible, at least in part, for their pathogenesis. This study was aimed at verifying whether chronic administration of cabergoline, a long acting dopamine D2-like receptor agonist, can reverse established LID. Four MPTP treated cynomolgus monkeys with long-standing and stable parkinsonian syndrome and reproducible dyskinesias to L-Dopa, were used in this study. We compared the antiparkinsonian and dyskinetic responses of L-Dopa methyl ester (62.5 mg and 125 mg), given with benserazide (50 mg) (L-Dopa/benserazide), administered before and after a 6-week period during which the animals were treated only by daily administration of cabergoline (doses ranging from 0.125 to 0.185 mg/kg, subcutaneous). During cabergoline treatment, the monkeys initially showed marked dyskinesias, which were reduced significantly after 4 weeks of treatment. However, there was no tolerance to its antiparkinsonian effect. L Dopa/benserazide given 4 days after cabergoline withdrawal produced a significant antiparkinsonian effect, but dyskinesias were dramatically reduced compared to what had been seen before chronic cabergoline treatment. The duration of the L Dopa response was not increased after chronic administration of cabergoline. Our data suggest that sustained dopamine D2 receptor stimulation could be of value when trying to reduce or to reverse LID in patients with fluctuating advanced PD. PMID- 11020124 TI - A pharmacoeconomic evaluation of botulinum toxin in the treatment of spasmodic torticollis. AB - We performed a prospective study in 21 patients to evaluate the cost of treatment of spasmodic torticollis (cervical dystonia) before and after botulinum toxin type A (BTA) treatment and to assess the impact of BTA treatment on quality of life. Data were recorded for the analysis over a period starting 8 months before and ending 7.2 +/- 0.2 months (mean +/- SEM) after the first injection of BTA. All patients received at least two BTA injections (2.9 +/- 0.2 injections per patient). We studied direct medical costs (drugs, outpatient and inpatient visits, diagnostic procedures, physiotherapy), clinical effects of BTA (clinical rating scale and patient's global assessment), quality of life (French version of the Nottingham Health Profile [NHP]), and adverse reactions. Costs associated with the treatment of spasmodic torticollis before the first BTA injection were 479 +/- 143 French Francs (FF)/patient/month (97 +/- 29 US $/pt/mo). During BTA treatment, costs were 1,126 +/- 147 FF/pt/mo (228 +/- 30 US $/pt/mo), including a mean cost of BTA of 771 +/- 131 FF/pt/mo (157 +/- 27 US $/pt/mo). Treatment with BTA significantly decreased clinical symptoms of spasmodic torticollis and improved the emotional, social, and pain-related domains of the quality of life assessment. Botulinum toxin type A treatment increases the cost of treating spasmodic torticollis but improves quality of life in terms of pain, social, and psychologic functioning in patients with spasmodic torticollis. PMID- 11020125 TI - Effect and time course of deep brain stimulation of the globus pallidus and subthalamus on motor features of Parkinson's disease. AB - We studied the effect and temporal profile of deep brain stimulation (DBS) of the globus pallidus and subthalamic nucleus on the motor signs of Parkinson's disease (PD). Four patients with bilateral deep brain stimulators of the globus pallidus and four patients with bilateral deep brain stimulators of the subthalamus were studied while taking no medication and at 15 and 30 minutes and 1, 2, 4, and 6 hours after turning stimulation on. An immediate (15 minutes) and sustained (6 hours) benefit was observed for all the motor manifestations of PD for both stimulation sites. Deep brain stimulation of the globus pallidus and subthalamus is highly effective in reducing all the cardinal motor features of PD. PMID- 11020126 TI - Serum vitamin B6 in schizophrenic and schizoaffective patients with and without tardive dyskinesia. AB - There are several reports regarding the efficacy of vitamin B6 in the treatment of tardive dyskinesia (TD). Vitamin B6 plays a key role in the synthesis of several neurotransmitters, including serotonin, dopamine, norepinephrine, and gamma-aminobutyric acid, all of which have been proposed to be involved in the development of TD. The purpose of this study was to examine whether there are special markers to distinguish long-term neuroleptic exposure patients who have TD from those patients who do not develop this side effect. In view of the pivotal role of vitamin B6 in the synthesis of all neurotransmitters believed to take part in the pathogenesis of TD, we decided to examine whether basal levels of vitamin B6 might explain the difference between these two groups. Such a finding could provide a predictive marker for vulnerable patients. The active metabolite of vitamin B6 is pyridoxal phosphate (PP). Pyridoxal phosphate blood levels were measured in 15 schizophrenic and schizoaffective patients with TD and compared with 15 patients without evidence of TD (matched by sex, age, smoking, and diagnosis). We found that, although patients in the TD group were exposed to neuroleptic drugs for significantly longer periods of time, there were no differences in serum PP levels between the groups. The reports of the effectiveness of vitamin B6 supplementation in the treatment of TD could therefore be explained by the assumption that central nervous system or intracellular vitamin B6 levels, which are involved in the pathogenesis of TD, are not the same as vitamin B6 peripheral serum levels. There is need for further studies, which will clarify the relationship between vitamin B6 and TD. PMID- 11020128 TI - Reversible pitch perception deficit caused by carbamazepine. AB - Carbamazepine (CBZ) is an antiepileptic drug frequently used to treat a variety of neurologic diseases or symptoms. In addition, the drug is used as a mood stabilizer in patients with affective or schizophrenic disorders. Among its adverse effects, auditory disturbance is described rarely. In this report, we describe a 25-year-old woman who noted falsely higher pitch perception after starting CBZ treatment for schizoaffective disorder. We also review the literature reporting CBZ-associated abnormal pitch perception. PMID- 11020127 TI - Protease inhibitor-induced carbamazepine toxicity. AB - Neurologic manifestations of HIV infection are quite diverse and can develop into seizures. Because new drug therapies have been developed, it is important to know the interactions between antiretroviral and antiepileptic agents. A 36-year-old patient with HIV developed a set of progressive left hemiparesis and secondarily generalized partial seizures related to progressive multifocal leukoencephalopathy. Phenytoin and carbamazepine were necessary to control the seizures. Instead of diverse antiretroviral therapies, the viral load was increased. Protease inhibitors (ritonavir and saquinavir) were added to the treatment and the patient developed progressive ataxia related to carbamazepine toxicity. Carbamazepine was discontinued and the patient remained asymptomatic. The patient was diagnosed with carbamazepine toxicity related to the introduction of ritonavir. Ritonavir is a potent inhibitor of hepatic cytochrome P450, mainly the CYP3A4 isoform. Carbamazepine is metabolized by this subsystem. Ritonavir acted as a CYP3A4 inhibitor, diminishing carbamazepine metabolism and provoking an increase in serum levels and clinical toxicity. We present a case of interaction between ritonavir and carbamazepine. Interaction between antiepileptic and antiretroviral agents is an emergent problem caused by the increasing association of the two therapies. We recommend strict monitoring of serum antiepileptic drug (AED) levels to avoid toxicity and inadequate seizure control. PMID- 11020129 TI - Nefazodone in the adjunctive therapy of schizophrenia: an open-label exploratory study. AB - Compared to conventional antipsychotic medications, atypical antipsychotic medications demonstrate greater central serotonin (5HT2) receptor antagonism than dopamine type 2 (D2) receptor antagonism. Nefazodone, an antidepressant medication, exhibits 5HT2 receptor antagonism; we therefore wondered if its addition to stable regimens of antipsychotic medication would increase antipsychotic efficacy, independently of a primary effect on mood, through the mechanism of augmented 5HT2 receptor antagonism. In a pilot investigation, we administered nefazodone (400 mg/d) for 6 weeks as an open-label adjunct to antipsychotic medication in 10 patients with chronic schizophrenia. The patients were moderately depressed at baseline but did not meet criteria for major depressive episode. The Brief Psychiatric Rating Scale (BPRS) and Montgomery Asberg Depression Rating Scale scores showed statistically significant and clinically robust improvements with nefazodone treatment, which were maintained at follow-up evaluation 2 weeks after the end of nefazodone treatment. There were no adverse events. These results suggest that nefazodone may be a safe and effective adjunct to antipsychotic medications in schizophrenia and that augmentation of 5HT2 antagonism may prove to be a viable strategy for "boosting" antipsychotic efficacy and for treating depressive symptoms in schizophrenia. PMID- 11020130 TI - Polyradiculoneuritis after botulinum toxin therapy for cervical dystonia. AB - A 40-year-old man with cervical dystonia developed an acute inflammatory demyelinating polyradiculoneuritis after botulinum toxin type A treatment. Some cases of idiopathic brachial plexopathy and polyradiculoneuritis have been reported to date. Although a causal relationship is not firmly established, the clinical temporal profile suggests a pathogenic relationship. In patients with cervical dystonia, further use of type A botulinum toxin should be considered contraindicated, and the use of another type of botulinum toxin should be taken into consideration. PMID- 11020131 TI - Acute dystonia with quetiapine. PMID- 11020132 TI - Re: The apolipoprotein E epsilon4 allele increases the risk of drug-induced hallucinations in Parkinson's disease. PMID- 11020133 TI - Clinical pharmacokinetics of teicoplanin. AB - The glycopeptide antibacterial teicoplanin has become increasingly popular in the last decade with the rise in infections related to methicillin-resistant Staphylococcus aureus. Teicoplanin has 6 major and 4 minor components. It is predominantly (90%) bound to plasma proteins. Of the several methods available to measure concentrations in serum, fluorescence polarisation immunoassay has high reliability and specificity. Teicoplanin is not absorbed orally, but intravenous and intramuscular administration are well tolerated. Teicoplanin is eliminated predominantly by the kidneys and only 2 to 3% of an intravenously administered dose is metabolised. Total clearance is 11 ml/h/kg. Steady state is reached only slowly, 93% after 14 days of repeated administration. Elimination is triexponential, with half-lives of 0.4 to 1.0, 9.7 to 15.4 and 83 to 168 hours. Volumes of distribution are 0.07 to 0.11 (initial phase), 1.3 to 1.5 (distribution phase) and 0.9 to 1.6 (steady state) L/kg. A standard dosage regimen of 6 mg/kg every 12 hours for 3 doses, then daily, will produce therapeutic serum concentrations of > or = 10 mg/L in most patients. Higher dosages may be required in certain patients, for example intravenous drug abusers or those with burns, because of unpredictable clearance. Concentrations in bone reach 7 mg/L at 12 hours after a dose of teicoplanin 6 mg/kg, but reach only 3.5 mg/L in the cartilage. Doses of 10 mg/kg are necessary to achieve adequate bone concentrations. There is little penetration into cerebrospinal fluid or the aqueous or vitreous humour. In fat, concentrations may be subtherapeutic (0.5 to 5 mg/L) after a dose of 400mg. A single prophylactic dose of 12 mg/kg is sufficient to maintain therapeutic concentrations during cardiopulmonary bypass or burns surgery. High loading doses reduce the delay to attaining therapeutic concentrations. Premature neonates require a loading dose of 15 mg/kg and a maintenance dosage of 8 mg/kg daily to ensure therapeutic serum concentrations. Children need loading with 10 mg/kg every 12 hours for 3 doses followed by maintenance with 10 mg/kg/day. Clearance is reduced predictably in renal failure, and dosage adjustments can be based on the ratio of impaired clearance to normal clearance. In patients on haemodialysis, 3 loading doses of 6 mg/kg at 12-hour intervals followed by maintenance doses every 72 hours produced trough plasma concentrations of 8 mg/L in most patients at 48 hours. The monitoring of serum concentrations is not necessary to avoid toxicity, but can be helpful in certain patient groups to ensure therapeutic concentrations are present, especially in those not responding to treatment. PMID- 11020134 TI - Comparative pharmacokinetics of the carbapenems: clinical implications. AB - During the last few decades, several carbapenems have been developed. The major characteristic of the newer drugs, such as MK-826, is a prolonged half-life. Alternatively, some carbapenems have been developed that can be given orally, such as CS-834 and L-084. Although imipenem and panipenem have to be administered with a co-drug to prevent degradation by the enzyme dehydropeptidase-1 and reduce nephrotoxicity, the newer drugs such as meropenem, biapenem and lenapenem are relatively stable towards that enzyme. Structural modifications have, besides changes in pharmacology, also led to varying antimicrobial properties. For instance, meropenem is relatively more active against Gram-negative organisms than most other carbapenems, but is slightly less active against Gram-positive organisms. Except for half-life and bioavailability, the pharmacokinetic properties of the carbapenems are relatively similar. Distribution is mainly in extracellular body-water, as observed both from the volumes of distribution and from blister studies. Some carbapenems have a better penetration in cerebrospinal fluid than others. In patients with renal dysfunction, doses have to be adjusted, and special care must be taken with imipenem/cilastatin and panipenem/betamipron to prevent accumulation of the co-drugs, as the pharmacokinetic properties of the co-drugs differ from those of the drugs themselves. However, toxicity of the co drugs has not been shown. The carbapenems differ in proconvulsive activity. Imipenem shows relatively the highest proconvulsive activity, especially at higher concentrations. Pharmacodynamic studies have shown that the major surrogate parameter for antimicrobial efficacy is the percentage of time of the dosage interval above the minimum inhibitory concentration (MIC). The minimum percentage percentage of time above the MIC (TaM) needed for optimal effect is known in animals (30 to 50%), but not in humans. It is probably less than 100%, but may be higher than 50%. Dosage regimens currently in use result in a TaM of about 50% at 4 mg/L, which is the current 'susceptible' breakpoint determined by the National Committee for Clinical Laboratory Standards (NCCLS) for most micro organisms. Dosage regimens in patients with reduced renal clearance should be based on the TaM. The increased half-life of the newer carbapenems will probably lead to less frequent administration, although continuous infusion may still be the optimal mode of administration for these drugs. The availability of oral carbapenems will have a profound effect on the use of carbapenems in the community. PMID- 11020135 TI - Drug-Drug interactions of clinical significance in the treatment of patients with Mycobacterium avium complex disease. AB - Therapeutic and prophylactic regimens directed specifically against Mycobacterium avium complex (MAC) are increasingly being used in patients infected with the human immunodeficiency virus (HIV). Several of the drugs used in the management of MAC have been associated with significant drug interactions involving the cytochrome P450 (CYP) enzyme system. This enzyme system is also highly influenced by other drugs used in the management of patients with HIV, particularly the protease inhibitors, non-nucleoside reverse transcriptase inhibitors (NNRTIs) and azole antifungals. This article reviews the published concentrations or subtherapeutic concentrations of other drugs have been described. In particular, concurrent use of rifabutin with clarithromycin or fluconazole has resulted in increased concentrations of rifabutin and an accompanying increase in the incidence of rifabutin toxicities, including uveitis and leucopenia. Similar results have been seen when rifabutin is combined with protease inhibitors or delavirdine. The macrolides, clarithromycin and azithromycin, have also been associated with significant drug interactions. Clarithromycin has a higher affinity for CYP than azithromycin and, thus, is more frequently associated with clinically significant drug interactions. Clarithromycin is an inhibitor of CYP and may result in toxic concentrations of other drugs metabolised by this enzyme system. Such interactions have been described with rifabutin and the statin lipid lowering agents. In addition, nevirapine and efavirenz have been shown to significantly reduce clarithromycin concentrations, whereas the protease inhibitors and delavirdine may increase clarithromycin concentrations. Other drugs used in the management of patients with MAC are not metabolised by CYP and thus have a lower incidence of interactions, although the absorption of ciprofloxacin may be impaired when it is given with products containing multivalent cations, such as didanosine. However, clinicians must remain vigilant for drug interactions when reviewing a patient's medication profile, keeping in mind both interactions that have been described in the literature and those that may be predicted based upon known pharmacokinetic profiles. PMID- 11020136 TI - Effects of obesity on pharmacokinetics implications for drug therapy. AB - Obesity is a worldwide problem, with major health, social and economic implications. The adaptation of drug dosages to obese patients is a subject of concern, particularly for drugs with a narrow therapeutic index. The main factors that affect the tissue distribution of drugs are body composition, regional blood flow and the affinity of the drug for plasma proteins and/or tissue components. Obese people have larger absolute lean body masses as well as fat masses than non obese individuals of the same age, gender and height. However, the percentage of fat per kg of total bodyweight (TBW) is markedly increased, whereas that chrome P450 isoforms are altered, but no clear overview of drug hepatic metabolism in obesity is currently available. Pharmacokinetic studies provide differing data on renal function in obese patients. This review analyses recent publications on several classes of drugs: antibacterials, anticancer drugs, psychotropic drugs, anticonvulsants, general anaesthetics, opioid analgesics, neuromuscular blockers, beta-blockers and drugs commonly used in the management of obesity. Pharmacokinetic studies in obesity show that the behaviour of molecules with weak or moderate lipophilicity (e.g. lithium and vecuronium) is generally rather predictable, as these drugs are distributed mainly in lean tissues. The dosage of these drugs should be based on the ideal bodyweight (IBW). However, some of these drugs (e.g. antibacterials and some anticancer drugs) are partly distributed in adipose tissues, and their dosage is based on IBW plus a percentage of the patient's excess bodyweight. There is no systematic relationship between the degree of lipophilicity of markedly lipophilic drugs (e.g. remifentanil and some beta-blockers) and their distribution in obese individuals. The distribution of a drug between fat and lean tissues may influence its pharmacokinetics in obese patients. Thus, the loading dose should be adjusted to the TBW or IBW, according to data from studies carried out in obese individuals. Adjustment of the maintenance dosage depends on the observed modifications in clearance. Our present knowledge of the influence of obesity on drug pharmacokinetics is limited. Drugs with a small therapeutic index should be used prudently and the dosage adjusted with the help of drug plasma concentrations. PMID- 11020137 TI - Pharmacokinetics of etonogestrel and ethinylestradiol released from a combined contraceptive vaginal ring. AB - OBJECTIVE: To assess the pharmacokinetics of etonogestrel and ethinylestradiol released from a novel combined contraceptive vaginal ring (NuvaRing) releasing etonogestrel 120microg and ethinylestradiol 15 microg per day and compare them with those of a combined oral contraceptive containing desogestrel 150 microg/ethinylestradiol 30 microg (DSG/EE COC). DESIGN AND SETTING: This was a nonblind, randomised, crossover study in 16 healthy women. METHODS: All volunteers received one cycle of DSG/EE COC before being randomised to 1 of 2 treatment groups. The participants in group 1 received 1 cycle of DSG/EE COC, a treatment period with NuvaRing and an intravenous bolus injection of etonogestrel/ethinylestradiol (150 microg/30 microg). Those in group 2 received a NuvaRing treatment period, 1 cycle of DSG/EE COC and the same intravenous bolus injection. RESULTS AND CONCLUSIONS: After the insertion of NuvaRing, maximum serum concentrations of etonogestrel and ethinylestradiol were achieved in approximately 1 week. The concentrations subsequently showed a gradual linear decrease in time. The maximum serum concentrations of etonogestrel and ethinylestradiol were approximately 40 and 30%, respectively, of those for the DSG/EE COC. In comparison with the DSG/EE COC, the absolute bioavailability for NuvaRing was higher for etonogestrel (102.9 vs 79.2%) and similar for ethinylestradiol (55.6 vs 53.8%). Taking the difference in daily doses into account, systemic exposure to etonogestrel was similar for NuvaRing and the DSG/EE COC, whereas systemic exposure to ethinylestradiol with NuvaRing was only approximately 50% of that for the DSG/EE COC. PMID- 11020138 TI - Effect of symptom information and validity scale information on the malingering of depression on the MMPI-2. AB - MMPI-2 (Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) data from college students who were attempting to malinger depression (199 women and 171 men) were compared to MMPI-2 data from students who responded honestly (50 women and 45 men). Mean MMPI-2 scores were compared with analyses of variance, and students' success in malingering depression was evaluated with criteria based on cutting scores for validity indexes and on the clinical scales commonly associated with depression. Students who were given information about the validity scales or about the symptoms of depression were more successful at malingering than students who received no information, indicating that malingerers of depression may be able to elude detection by the MMPI-2 if they are informed about the validity scales or the symptoms of depression. PMID- 11020139 TI - Differential roles of depressive and anxious symptoms and gender in defensiveness. AB - Among a sample of Air Force cadets facing the prospect of basic training (N= 1,190; 1,005 men and 185 women), the influence of a defensive test-taking style on measures of depressive and anxious symptoms was examined. Participants completed the Beck Depression Inventory (Beck & Steer, 1987) and the Beck Anxiety Inventory (Beck, Epstein, Brown, & Steer, 1988), as well as the MMPI (Hathaway & McKinley, 1943) L scale. Results supported hypotheses that defensiveness would affect a self-report measure of depression but not a self-report measure of anxiety and would do so more among men than women. Applied implications of the results are discussed. PMID- 11020140 TI - Rorschach correlates of sexual abuse: trauma content and aggression indexes. AB - This study compared Rorschach records of nondissociative outpatients with histories of (a) definite sexual abuse (n = 22), (b) suspected but unconfirmed sexual abuse (n = 13), or (c) no sexual abuse (n = 43) on selected variables hypothesized to be associated with sexual abuse. As predicted, clients with definite sexual abuse scored significantly higher than clients known not to be sexually abused on Armstrong and Loewenstein's (1990) Trauma Content index (TC/R), with an effect size greater than 1 SD. Contrary to prediction, there was no significant difference in the frequency of their Aggressive Past (AgPast; Gacono & Meloy, 1994) scores. AgPast scores, however, did positively correlate with sexual abuse that was violent or sadistic. As a test of discriminant validity, we hypothesized that 2 Rorschach variables (PER and Sc) would be unrelated to sexual abuse. This was supported by our data. Although TC/R was strongly associated with the presence and severity of sexual abuse, it could not discriminate sexually abused from nonsexually abused clients with great accuracy. The TC/R score is 1 factor among many that can be used to assess the validity of clients' claims of past sexual abuse. PMID- 11020141 TI - Reliability of interview-assessed hostility ratings across mode of assessment and time. AB - These studies addressed 2 questions concerning interview-based hostility assessments: whether they are affected if the interview is conducted face-to-face versus telephone and whether they are stable across an extended time period. In Study 1A, 54 students were interviewed face-to-face and by telephone in a laboratory setting. Half the sample was reinterviewed in the laboratory 6 weeks later. The other half was reinterviewed by telephone at home. With 1 exception, all intraclass correlation coefficients (ICCs) comparing interview modes were above .62. In Study 1B, 48 adults were interviewed face-to-face in a laboratory and by telephone in their homes with a 2-week intervening interval. The ICC comparing interview modes was .78. In Study 2, 100 adults were interviewed face to-face in a laboratory and approximately 4 years later by telephone in their homes. The ICC across interviews was .69. Thus, support was found for the stability of interview-based hostility assessments across interview methods and extended periods of time. PMID- 11020142 TI - Personality characteristics of ADHD adults assessed with the Millon Clinical Multiaxial Inventory-II: evidence of four distinct subtypes. AB - This study compared the personality characteristics of 104 adults diagnosed with attention deficit hyperactivity disorder (ADHD). Personality features were assessed with the MCMI-II (Millon, 1987). Participants were divided into 4 groups based on the presence of persisting oppositional defiant disorder (ODD) or other comorbid diagnoses (ADHD only, ADHD-comorbid, ADHD-ODD, ADHD-ODD-comorbid). Significant differences between these groups were present for 9 of the 13 MCMI-II personality scales, resulting in 4 modal personality styles. ADHD-only adults evidenced mild histrionic traits, whereas the ADHD-comorbid group was more often avoidant and dependent in personality style. ADHD-ODD adults showed histrionic, narcissistic, aggressive-sadistic, and negativistic traits whereas the ADHD-ODD comorbid group had a combination of avoidant, narcissistic, antisocial, aggressive-sadistic, negativistic, and self-defeating personality features. Implications for treatment are discussed. PMID- 11020143 TI - Circumplex scales of interpersonal values: reliability, validity, and applicability to interpersonal problems and personality disorders. AB - Three studies describe the development, psychometric properties, and potential utility of a new self-report measure, the Circumplex Scales of Interpersonal Values (CSIV). The CSIV was designed to complement other interpersonal circumplex measures that assess interpersonal behavior by efficiently assessing a comprehensive set of agentic and communal values. The eight 8-item scales of the CSIV were shown to have good internal consistency and test-retest reliability and a circumplex structure. The CSIV showed convergent and discriminant validity with measures of interpersonal traits (the Bem Sex Role Inventory; Bem, 1974), interpersonal problems (the Inventory of Interpersonal Problems-Circumplex; Horowitz, 2000), implicit interpersonal motives (the Thematic Apperception Test; see Atkinson, 1958), and interpersonal goals (the Interpersonal Goals Inventory; Dryer & Horowitz, 1997). Finally, the locations of the MCMI-III (Millon, 1994) personality disorder scales on the CSIV circumplex generally mirrored the locations of personality disorders on other interpersonal circumplex measures. PMID- 11020144 TI - Utility of the MMPI-2 in assessing gender dysphoric patients. AB - We compared MMPI-2 (Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989) profiles of 2 groups of adult biological men requesting sex reassignment surgery; 1 group was diagnosed with Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev. [DSM-III-R]; American Psychiatric Association, 1987) transsexualism and the other with gender identity disorder of adolescence and adulthood, nontranssexual type (GIDAANT). Although the mean profiles for the transsexual group did not demonstrate any psychopathology, the GIDAANT group showed moderate psychopathology. A cluster analysis indicated that 85% of the transsexual group showed low psychopathology and 47% of the GIDAANT group showed severe psychopathology. Neither the MMPI-2 results nor the DSM-III-R clinical evaluation support the conclusion of many authors that transsexualism is associated with severe personality disorder; rather, the data indicate that transsexualism and other gender identity disorders without persistent wish for sex reassignment differ significantly in degree of psychopathology. PMID- 11020145 TI - A study of nonpatient preadolescent Rorschach protocols. AB - There is a paucity of peer-reviewed, nonpatient Rorschach studies in psychological journals, particularly for children. This study examined 100 preteenage children, using the Comprehensive System to code their Rorschach protocols. The participants were also evaluated with the Conners Parent Rating Scale-93 (Conners, 1989) and 5 exclusionary behavior criteria. The 100 participants demonstrated better than average behaviors. The results presented include all the ratios and indexes found in the nonpatient statistics for the Comprehensive System. We discuss interrater reliability issues and offer recommendations for further cross-validating research. PMID- 11020146 TI - Comparing the relative fit of various factor models of the self-consciousness scale in two independent samples. AB - Research shows that using highly self-aware participants yields studies of higher reliability, validity, and statistical power; dispositional self-awareness is commonly measured using the Fenigstein Self-Consciousness Scale (Fenigstein, Scheier, & Buss, 1975). This study used confirmatory factor analysis to compare various factor models that may underlie that scale. Two independent student samples (296 from Bernstein, Teng, & Garbin, 1986, and 350 from a large Canadian university) completed the scale. Using 6 fit criteria, results from both samples supported the Burnkrant and Page (1984) 4-factor model, namely, that self consciousness consists of 3 principle scales: Social Anxiety, Public Self Consciousness, and Private Self-Consciousness (divided into Internal State Awareness and Self-Reflectiveness). We discuss the psychometric implications of enhancing scale reliability, validity, and self-awareness. PMID- 11020147 TI - Development and validation of a scale to measure perceived control of internal states. AB - One of the key developments in the psychological literature on control has been the growing recognition of the multidimensional nature of the control construct. Recent research suggests that perceived control of internal states may be just as important as perceived control of external events. The Perceived Control of Internal States Scale was developed to provide a measure of the degree to which people feel they have control of their internal states (emotions, thoughts, physical reactions). I report the results of 2 studies (N= 689), supporting the reliability, construct, and incremental validity of the scale. The buffering effects of perceived control for people facing major life events was also explored, with higher levels of perceived control being associated with less physical and psychological symptoms of strain. PMID- 11020148 TI - Feigning combat-related posttraumatic stress disorder on the personality assessment inventory. AB - This study examined whether individuals who were instructed on the Diagnostic and Statistical Manual of Mental Disorders (4th ed. [DSM-IV]; American Psychiatric Association, 1994) criteria for posttraumatic stress disorder (PTSD) could feign PTSD on the Personality Assessment Inventory (PAI; Morey, 1991). The study also investigated whether PAI indexes of symptom exaggeration, the Negative Impression Management (NIM) scale and the Malingering index, could identify individuals feigning PTSD. The diagnostic rule for PTSD (Morey, 1991, 1996) was applied to the profiles of a group of 23 veterans with combat-related PTSD and 23 male undergraduates instructed to malinger PTSD. Seventy percent of the student malingerers produced profiles that received diagnostic consideration for PTSD. The NIM cutting score (> or = 8) was highly effective in detecting simulation of PTSD but resulted in the misclassification of a large number of true PTSD cases. There were no significant differences in the overall efficiency of the test with various validity criteria. We discuss the implications of these findings for the use of the PAI in the diagnosis of combat-related PTSD. PMID- 11020149 TI - Feast and potential famine. PMID- 11020150 TI - Misuse of the term "containment" in laboratory animal science. PMID- 11020151 TI - National Institutes of Health--supported opportunities in Comparative Medicine. AB - The Comparative Medicine (CM) area of the National Institutes of Health (NIH) is a major source of support for research on laboratory animals, training of laboratory animal specialists, and support of shared, regional animal resources. We present a brief history of CM at NIH and the major mechanisms by which it accomplishes its goals in programs located across the United States. PMID- 11020152 TI - Partial hepatectomy and laparoscopic-guided liver biopsy in rhesus macaques (Macaca mulatta): novel approach for study of liver regeneration. AB - BACKGROUND AND PURPOSE: Although valuable information has been gained using a rodent partial hepatectomy model to assess liver regeneration, the ability to apply this research to humans remains uncertain. Thus, liver regeneration was assessed in a non-human primate, the rhesus macaque (Macaca mulatta). METHODS: One animal underwent 60% hepatectomy, a second animal underwent 30% hepatectomy, and control surgery (cholecystectomy) was performed on two separate animals. Laparoscopic-guided liver biopsy was performed on days 1, 2, 7, 14, and 30 after surgery. Changes in hemoglobin concentration and alanine transaminase activity were assessed, and liver regeneration was evaluated by measuring the expression of Ki-67. RESULTS: All animals survived surgery and laparoscopy. Substantial liver regeneration was induced in the animal that underwent 60% hepatectomy. Excellent tissue specimens were obtained via laparoscopic-assisted liver biopsy. CONCLUSIONS: Sixty percent partial hepatectomy in rhesus macaques appears to be an excellent model for the study of hepatocellular regeneration. The procedure was safe, and effectively induced liver regeneration. In addition, laparoscopic guided liver biopsy allows observation of changes in the liver remnant as regeneration develops, and provides excellent tissue specimens for analysis. Thus, this rhesus macaque partial hepatectomy model will allow further characterization of liver regeneration in a species closer to humans. PMID- 11020153 TI - An improved genetic linkage map of rat chromosome 20. AB - BACKGROUND AND PURPOSE: Rat chromosome 20 is one of special interest because it contains some diabetogenic genes, such as a major histocompatibilitiy complex (MHC)-linked genetic components and quantitative trait loci. We studied rat chromosome 20, using the backcross progeny between BB/Wor and PVG.R23 rats, and confirmed the genetic linkage map by use of another backcross panel. METHODS: Backcross panels were done between BB/Wor and PVG.R23 rats, and BN and KZC rats. Length variations of simple sequence length polymorphism markers were analyzed by use of polymerase chain reaction (PCR) analysis. Alleles of RT1-Bb and RT1-Db were analyzed by use of the PCR-restriction fragment length polymorphism method. Genetic maps of rat chromosome 20 were constructed, using the Map Manager computer program. RESULTS: Fifty-two loci were mapped on rat chromosome 20. Genetic length was 57.9 cM, with average spanning of 1.11 cM between markers. The positions of RT1-N1, Tnf, and RT1-Bb into the MHC region were separated and confirmed by results of two backcross panels in our linkage studies. CONCLUSIONS: The genetic linkage map of rat chromosome 20 was improved, and was a useful tool for genetic analysis of a diabetogenic gene(s) and for producing MHC congenic strains. PMID- 11020154 TI - Guinea pig abscess/hypersensitivity model for study of adverse vaccination reactions induced by use of Q fever vaccines. AB - BACKGROUND AND PURPOSE: The Coxiella burnetii phase-I cellular vaccine is efficacious in humans, imparting nearly complete protection against Q fever. However, this vaccine can also induce sterile abscesses and granulomas at the inoculation site in humans previously sensitized by natural infection or vaccination. To decrease the possibility of vaccinating immune persons, vaccinees are currently screened by skin testing to detect pre-existing Q fever immunity. We developed a model of abscess hypersensitivity in Hartley guinea pigs to assess the likelihood that Q fever vaccines would induce adverse vaccination reactions in previously sensitized individuals. METHODS: Guinea pigs (4 to 6/group) were sensitized to C. burnetii by immunization and aerosol challenge, or by intraperitoneal inoculation. Eight weeks later, animals were then vaccinated SC with a Q fever cellular (WCI) or chloroform:methanol residue (CMR) vaccine. Development of adverse reactions at the vaccination site was assessed histologically and by observation of increases in erythema and/or induration. RESULTS: The WCI vaccine caused greater magnitude and duration of erythema and induration at the vaccination sites than did the CMR vaccine. In addition, non immune guinea pigs developed induration when given WCI, but not CMR vaccine. CONCLUSIONS: The CMR vaccine may prove a safe alternative to WCI vaccines for use in individuals unscreened for prior immunity to C. burnetii. PMID- 11020155 TI - Ultrasound measurement of biparietal diameter and umbilical artery blood flow in the normal fetal guinea pig. AB - BACKGROUND AND PURPOSE: Measurement of the biparietal diameter (BPD) by use of B mode ultrasound provides a useful means for assessment of gestational age and brain growth in humans during pregnancy. Recording of flow velocity waveforms from the umbilical artery, using Doppler ultrasound, is used to assess development of the fetal placental circulation. We sought to measure these ultrasound parameters during normal pregnancy in the guinea pig and develop normative data. METHODS: Measurements of BPD were made on 205 fetuses of various gestational ages; 114 fetuses had 2 or more serial studies performed (total n = 474). RESULTS: BPD increased from 0.806cm at 22 to 26 days, to 1.922cm at term (69 days), (y = -0.00043x2 + 0.06881x - 0.75941, with an r value of 0.995, where x = days' gestation, y = biparietal diameter [cm]). Umbilical artery flow velocity waveform resistance index (RI) decreased as gestation advanced (y = 0.012x + 1.294 with an r value of 0.887, where x = days gestation, y = RI) reflecting expansion of the placental vascular bed. CONCLUSIONS: It is possible to use ultrasound to study pregnancy in the guinea pig. The BPD may be used to estimate gestational age. Resistance to blood flow in the placenta may be assessed using the RI derived from the umbilical artery flow velocity waveform. PMID- 11020156 TI - Model of chronic systolic and diastolic dysfunction after cryothermia-induced myocardial necrosis in rats. AB - BACKGROUND AND PURPOSE: Left ventricular dysfunction following myocardial infarction is the most important predictor of adverse prognosis. Novel treatment options in infarction require an appropriate experimental model with a standardized, hemodynamically relevant myocardial injury. We evaluated a cryoinjury model in rodents that allows quantitative analysis of systolic and diastolic dysfunction. METHODS: Anesthetized, orally intubated, and ventilated Lewis rats (n = 12) underwent sternotomy. Myocardial necrosis was induced by use of a standardized cryolesion to the obtuse margin of the left ventricle, freezing for 3 minutes to -160 degrees C. Left ventricular performance was analyzed at day 120 after cryoinjury. Sham-operated animals (n = 10) served as controls. RESULTS: Cryoinjured animals behaved normally and gained weight up to day 120. Average heart weight of cryoinjured animals significantly exceeded that of controls. Left ventricular systolic pressure and systolic, diastolic, and mean aortic pressures were lower 4 months after cryoinjury, whereas left ventricular end-diastolic pressure was significantly increased. Cryoinjured animals had reduced aortic blood flow, as well as impaired maximal left ventricular dP/dt during aortic occlusion and aortic occlusion-provoked peak systolic pressure. Analysis of maximal rates of isovolumic pressure decrease revealed significant reduction in peak negative dP/dt in cryoinjured animals. Finally, time constants of isovolumic pressure decline were significantly prolonged in cryoinjured animals. CONCLUSION: Standardized cryothermia induces a myocardial lesion that results in highly reproducible impairment of left ventricular performance 120 days after cryothermia. The model is ideally suited to test novel therapeutic strategies for myocardial dysfunction. PMID- 11020157 TI - Comparison of two porcine (Sus scrofa domestica) skin models for in vivo near infrared laser exposure. AB - BACKGROUND AND PURPOSE: The current safety standards for lasers operating in the 1,400- to 2,000-nanometer (nm) wavelength region are based on only a few observations at specific wavelengths. On the basis of experimental results conducted with Yorkshire pigs (Sus scrofa domestica), these standards may not accurately reflect the potential for laser injury when humans are exposed to these wavelengths. It is our belief that one of the damage mechanisms involved in these laser injuries results from energy absorption by skin pigmentation (melanin), and a more highly pigmented animal model, the Yucatan hairless minipig, may be a more suitable subject for laser exposure studies. METHODS: Skin specimens were collected from Yorkshire pigs and Yucatan minipigs for histologic examination, and the thickness of the epidermis was measured. Epidermal thickness of human skin also was determined, and a qualitative assessment of the melanin content in the epidermal layers was conducted. RESULTS: Mean +/- SD thicknesses of the Yucatan minipig flank and dorsal neck epidermis were 68 +/- 34 and 68 +/- 25 microm, respectively. Thicknesses of the Yucatan minipig skin were closely comparable to the thicknesses of human epidermis from the face (68 +/- 26 microm), neck (65 +/- 24 microm) and arms (68 +/- 21 microm). The Yorkshire pig lacks substantial melanin in the epidermis, whereas the skin of the Yucatan minipig is more similar to that of humans. CONCLUSION: On the basis of epidermal skin thickness measurements and melanin assessment, the flank and dorsal neck of the Yucatan minipig are better suited to laser injury studies than are the Yorkshire pig models of human skin. PMID- 11020158 TI - Long-term infusion of nonphysiologic solutions into brain parenchyma: effects of pH, osmolarity, and flow rate. AB - The effects of long-term (3-day) infusion of nonphysiologic solutions into brain parenchyma were investigated in male Fischer (F344) 344 rats. Two weeks prior to infusion, a guide cannula was placed into the striatum, substantia nigra, or hippocampus. Solutions were infused continually for 3 days at flow rates of 0.03 (129.6 microl total) or 0.10 (432 microl total) microl/min. Four days after infusion, rats were euthanized and the brain was removed and processed for histologic evaluation. Rats that received cannula implants alone had the usual mechanical damage induced by implantation of the cannula. The brain regions that received 0.9% saline, pH 5.0 or pH 9.0 buffer at the two aforementioned flow rates had only minor evidence of tissue damage adjacent to the infusion site that was similar to that attributable to mechanical damage from the cannula implants. Brain tissue infused with distilled water or 1.8% saline also had modest effects of the solutions similar to the usual mechanical damage induced by the infusion cannulae. In contrast, contamination of the infusion sites was seen to induce inflammation. Data from these studies support the hypothesis that nonphysiologic solutions can be used to deliver compounds into brain parenchyma, without the infusion solutions themselves causing excess damage to brain tissue. PMID- 11020160 TI - Survey of embryonic stem cells for murine infective agents. PMID- 11020159 TI - Echocardiography, color-coded Doppler imaging, and abdominal sonography, a non invasive method for investigation of heart and aortic morphology and function in female gottingen minipigs: method and reference values for M-mode, B-mode, and flow parameters. AB - OBJECTIVE: The aim of the study reported here was to set up a method for echocardiography (EC) and abdominal sonography and to obtain EC reference values for left ventricular (LV) morphology and function and sonographic abdominal aortic morphology, function, and flow values in conscious, unsedated Gottingen minipigs. METHODS: Applying a standardized investigation procedure, the following parameters were measured by use of M-mode EC, color-coded Doppler imaging, and B mode sonography, or were calculated, in 58 female minipigs: LV end-diastolic and end-systolic diameter, interventricular septum thickness, LV caudal wall thickness, LV end-diastolic volume and end-systolic volume, fractional shortening, ejection fraction, and percentage of thickening of interventricular septum and LV caudal wall. In addition, morphology, pulsatility, flow values, and flow patterns in the abdominal aorta were recorded or calculated during abdominal sonography and color-coded Doppler imaging. RESULTS: Variable EC values were obtained due to individual variations of motor activity. Variation could be reduced by accustoming the animals to a standardized investigation procedure. Reference values could be obtained for EC, partially indicating clear correlation with body weight. Color-coded Doppler and Doppler spectra did not indicate flow disturbances in large arterial abdominal vessels. CONCLUSIONS: Results indicate that handling during EC and sonography can cause discomfort in unsedated minipigs that may interfere with recording of valid reference values for functional cardiac parameters in young animals. Accustoming the animals to a standardized investigation procedure reduces stress to a satisfactory level and enables data recording. Thus the minipig is considered suitable for assessment of cardiovascular parameters in experimental or toxicologic studies. PMID- 11020161 TI - Type 2 diabetes mellitus, hyperlipidemia, and extremity lesions in California mice (Peromyscus californicus) fed commercial mouse diets. AB - BACKGROUND AND PURPOSE: We characterized abnormalities of carbohydrate and lipid metabolism and determined whether those metabolic abnormalities are associated with extremity lesions in California mice (Peromyscus californicus). METHODS: Blood samples were evaluated for glucose, cholesterol, triglyceride, and insulin concentrations. Necropsy and histologic evaluation were done on selected mice, including staining pancreatic sections for insulin. Physical examinations also were performed. RESULTS: California mice were found to have Type 2 diabetes mellitus (T2DM). Sections of pancreas from diabetic and prediabetic mice had pathologic changes consistent with T2DM. After six months of feeding a low-fat diet, mice were normoglycemic, normotriglyceridemic, and normocholesterolemic. Some mice remained hyperinsulinemic. Traumatic lesions were not associated with T2DM. CONCLUSIONS: California mice develop diet-related T2DM when fed a diet containing 25.8% kcal from fat. California mice may be a useful animal model of human T2DM, and traumatic lesions result from housing California mice in multiple male groups. PMID- 11020162 TI - Use of guanfacine to control self-injurious behavior in two rhesus macaques (Macaca mulatta) and one baboon (Papio anubis). AB - BACKGROUND AND PURPOSE: Self-injurious behavior (SIB) affects 0.8 to 10% of individually housed non-human primates, and is a substantial threat to their health and well being. The potential for SIB to involve multiple neurotransmitters and the complex variations in response to external stressors complicate case management. Modulation of the adrenergic system by use of guanfacine, an alpha2A-adrenergic receptor agonist, was assessed as a novel therapeutic strategy for SIB. METHODS: The efficacy of guanfacine against SIB was evaluated in 11 self-biting episodes among two rhesus macaques (Macaca mulatta) and one baboon (Papio cynocephalus anubis). Affected animals were given guanfacine IM or PO at 0.5 mg/kg of body weight twice daily (rhesus) or 0.3 mg/kg (baboon) for 5 to 10 days, followed by gradual reduction of the dose to 0.25 mg/kg (rhesus) or 0.15 mg/kg (baboon) once daily over an average of 33 days. RESULTS: The 0.5 mg/kg twice daily regimen of guanfacine halted all self-biting, whereas reducing the dose to 0.25 mg/kg given twice daily or 0.5 mg/kg given once daily resulted in reversion to self-biting in four of the 11 episodes. Recurrence was controlled by returning to twice daily 0.5 mg/kg dosing for one aggressive episode, and resolved in the three milder episodes without dose or frequency being increased. Self-biting after discontinuation of therapy recurred six times over five years in case 1, three times over 1.5 years in case 2, and three times over one year in case 3. Clinical assessment suggested that guanfacine therapy decreased agitation without overt side effects associated with alpha2-agonists, such as profound sedation. CONCLUSION: The mechanism for guanfacine inhibition of self-biting is unclear, but could result from strengthening of prefrontal cortex inhibitory functions. Guanfacine therapy provides an effective psychological stabilizing tool that alleviates self-biting, and provides time to assess and address external stressors and triggers. PMID- 11020163 TI - Mousepox resulting from use of ectromelia virus-contaminated, imported mouse serum. AB - Mousepox was identified in a single mouse-holding room in early 1999 after a group of 20 CAF1/Hsd mice were inoculated SC with a killed murine spindle cell tumor line, S1509A. The cell line had been used without complications multiple times and was determined to be free of viral contamination on the basis of results of mouse antibody production testing. Of the 20 mice inoculated, 12 mice died by postinoculation day 8. Severe lymphoid and hepatic necrosis was observed in select mice subjected to histologic examination. Ballooning degeneration of epithelial cells with intracytoplasmic eosinophilic inclusion bodies was observed in the skin overlying the inoculation site of the single mouse from which this tissue site was evaluated. Presence of ectromelia virus was confirmed by use of immunohistochemical and polymerase chain reaction analyses, and the virus was isolated after serum, pooled from 5 of the index cases, was inoculated into an immune-naive mouse. Investigation into the source of virus contamination included inoculating mice with aliquots of various S1509A freeze dates; chemically defined media and supplements, including fetal bovine serum; and two lots of pooled commercial mouse sera, after heat inactivation at 56 degrees C for 30 minutes used as a medium supplement. One lot of pooled commercial mouse serum was identified as the source of ectromelia virus. This lot of serum was inadvertently used to feed S1509A cells that were subsequently inoculated into mice. We determined that the contaminated serum, which was purchased in late 1998, originated from China. The serum was imported into the United States as a batch of 43 L in early 1995. The serum was blended into a single lot and filtered (0.2 microm) before distribution to major suppliers throughout the country. The serum was sold or further processed to obtain a variety of serum-derived products. Because murine serum is generally sold in small aliquots (10 to 50 ml), we speculate that several thousand aliquots may have been derived from this batch of serum and, if inoculated into mice, would likely result in additional mousepox outbreaks. PMID- 11020164 TI - Monitoring sentinel mice for Helicobacter hepaticus, H rodentium, and H bilis infection by use of polymerase chain reaction analysis and serologic testing. AB - BACKGROUND AND PURPOSE: Natural infection of research mice with enterohepatic Helicobacter spp. is common and may confound experimental studies from intercurrent disease. We evaluated a protocol of dirty bedding exposure for transmission of Helicobacter infection from colony mice to female Tac:(SW)fBR sentinel mice over 6 months. METHODS: Cecal scrapings from culled colony mice and associated sentinel mice were tested for H. hepaticus, H. rodentium, and H. bilis using polymerase chain reaction analysis (PCR). These results were correlated with the results of sentinel serum IgG responses measured by ELISA. RESULTS: In 9 colony rooms, 43 of 45 mice were infected with H. hepaticus; in 14 rooms, 58 of 70 mice were infected with H. rodentium; and in 2 rooms, 2 of 10 mice were infected with H. bilis. Concurrence of Helicobacter infection between colony and sentinel mice was 82% for H. hepaticus, 88% for H. rodentium, and 94% for H. bilis. Concurrence of Helicobacter infection status of sentinel cagemates was 98% for H. hepaticus, 86% for H. rodentium, and 95% for H. bilis. Fecal samples pooled by sentinel cage had positive PCR results for H. hepaticus and H. rodentium at 1 month in 60 and 44%, respectively, of the cages that contained test-positive mice at necropsy (6 months). By 3 months, detection rates were 100 and 81% for H. hepaticus and H. rodentium, respectively, and H. bilis was not detected until 4 months. Newly acquired infections with H. rodentium and H. bilis were evident throughout the 6-month study period. Seroconversion was coincident with positive PCR results in sentinel mice, and serum IgG values continued to increase until necropsy. The serum IgG ELISA was 98 to 100% sensitive, but was low in specificity (34 to 44%), most likely attributable to common coinfection with H. hepaticus and H. rodentium. CONCLUSION: Sentinel mice acquire infection with Helicobacter spp. through dirty bedding exposure. Combined use of PCR analysis and serologic testing of sentinel mice was predictive of Helicobacter infection status of mouse colonies used for biomedical research. PMID- 11020165 TI - Eradication of infection with Helicobacter spp. by use of neonatal transfer. AB - BACKGROUND AND PURPOSE: Efficient methods for detection and elimination of Helicobacter spp. infections are needed to facilitate the development of Helicobacter-free mouse colonies. We developed an inexpensive, high-throughput method for preparation of fecal DNA for Helicobacter polymerase chain reaction (PCR) assays. METHODS: Fecal DNA was prepared by heating fecal pellets to 95 degrees C for 10 minutes in an alkaline solution, then adjusting the pH by addition of Tris buffer. This solution is used for PCR assays without purification of DNA. We then tested fostering as a method of generating Helicobacter-free mice. Litters born to Helicobacter-positive dams were transferred to Helicobacter-negative foster dams on the first day of life. RESULTS: Fostered pups tested Helicobacter negative up to 89 days of age, whereas pups raised by Helicobacter-positive dams were all test positive by 19 days of age. CONCLUSION: These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies. PMID- 11020167 TI - Otalaryngology-head and neck surgery: moving further into the electronic media PMID- 11020166 TI - Diverticulitis with rupture and fatal peritonitis in a Sumatran orangutan (Pongo pygmaeus). AB - A 30-year-old male Sumatran orangutan (Pongo pygmaeus) presented with signs of depression, lethargy, anorexia, and diarrhea that progressed to acute colic. Exploratory laparotomy revealed fibrinopurulent peritonitis and 50 cm of devitalized small intestine. The surgically resected small intestine contained several mucosal diverticula along the mesenteric attachment; one had ruptured, resulting in peritonitis. Fifteen days after surgery, the orangutan's abdominal incision dehisced. Repeated laparotomy revealed dehiscence of the distal intestinal anastomosis site, as well as extensive adhesions and purulent exudate. The defect was repaired, and the abdomen was extensively irrigated and closed, but the animal died within 24 hours. To our knowledge, this is the first report of diverticulitis in a great ape. Diverticulosis should be considered in the differential diagnosis for great apes that present with signs of depression, lethargy, anorexia, and/or diarrhea. PMID- 11020168 TI - Elevated nitric oxide metabolite levels in chronic sinusitis. AB - Decreased exhaled nitric oxide (NO) is found in chronic sinusitis. NO metabolites (nitrates, nitrites, and S-nitrosothiols) were measured in sinus lavages with a rabbit model of chronic sinusitis. NO metabolite levels (mean +/- SD) were 3.0+/ 1.6 micromol/L in uninfected rabbits, 10.7+/-11.4 micromol/L in infected animals, and 7.6+/-5.4 micromol/L in postantrostomy recovering animals. Infected sinuses had elevated levels of NO metabolites that were statistically significant (P<0.01) when compared with uninfected sinuses. Mucociliary transport velocity was measured in uninfected (16.0+/-5.7 mm/minute), infected (5.2+/-1.3 mm/minute), and recovery phases (3.0 mm/minute). Endoscopic appearance, light and electron microscopy, and bacterial cultures improved during recovery. Elevated levels of NO metabolites were found during chronic sinusitis and began to return to normal levels during recovery. The possible link between NO in epithelial autotoxicity and host defense mechanisms warrants further investigation. PMID- 11020169 TI - Analysis of aerobic bacterial strains found in chronic rhinosinusitis using the polymerase chain reaction. AB - INTRODUCTION: Rhinosinusitis is a common disease affecting an estimated 14% of the population. Although there is general agreement in the literature regarding acute rhinosinusitis, chronic rhinosinusitis is not as well studied, and no consensus has been reached regarding the bacterial etiology. The goal of this study was to test chronic rhinosinusitis mucosal specimens using the polymerase chain reaction (PCR) for aerobic bacterial content and to compare the results with standard culture data. RESULTS: Routine culture samples grew 50% aerobic bacteria, whereas PCR detected 62% aerobic bacteria contamination. CONCLUSION: PCR detected more bacteria in mucosal samples than in standard culture, but standard culture of this mucosa reflects the general aerobic bacteria found in chronic rhinosinusitis, with no predominant species of aerobic bacteria. SIGNIFICANCE: The analysis of chronic rhinosinusitis mucosa with the PCR method should give a more accurate picture of the bacteria found in chronic rhinosinusitis so that proper therapy can be instituted. PMID- 11020170 TI - Factors related to outcome of salvage therapy for isolated cervical recurrence of squamous cell carcinoma in the previously treated neck: a multi-institutional study. AB - OBJECTIVE: The goal was to identify factors associated with the outcome of salvage therapy for patients with isolated cervical recurrences of squamous cell carcinoma in the previously treated neck (ICR-PTN). STUDY DESIGN AND SETTINGS: A tumor registry search for ICR-PTN patients was performed at 7 participating institutions, and the charts were reviewed. Kaplan-Meier plots for survival and time until re-recurrence were used to evaluate the significance of associated variables. RESULTS: Median survival and time until re-recurrence were both 11 months. Survival was better in patients with the following characteristics: nonsurgical initial neck treatment, negative initial disease resection margins, no history of prior recurrence, ipsilateral location of the ICR-PTN relative to the primary, and use of surgical salvage. CONCLUSIONS: By pooling the experience of 7 US tertiary care medical centers, we have identified 5 factors that are associated with outcome of salvage therapy for ICR-PTN. SIGNIFICANCE: Consideration of these factors, as well as the reviewed literature, should facilitate patient selection for salvage protocols. PMID- 11020171 TI - Numerical densities of myonuclei and satellite cells in muscle fiber types in the aging human thyroarytenoid muscle: an immunohistochemical and stereological study using confocal laser scanning microscopy. AB - OBJECTIVE: This study determines the role of changes in numerical densities of myonuclei and satellite cells in age-related remodeling of the thyroarytenoid muscle (TA). DESIGN: Changes in numerical densities (N(V)) and ratios (N(N)) of myonuclei and satellite cells were estimated for the entire TA by use of stereological techniques. RESULTS: There was no age-related change or difference between fiber types for N(V myonucleus, fiber), but N(V myonucleus, fiber) increased with decreasing fiber diameter. There was a trend toward a decrease in N(V satellite cell, fiber) and a decrease in N(N satellite cell, myonucleus). N(V satellite cell, fiber) was higher for type 1 than for type 2 fibers, and type 1 satellite cells increased disproportionately with increasing total satellite cell numerical density. CONCLUSION: Decreased satellite cell proliferation may contribute to age-related fiber loss and atrophy in the TA. SIGNIFICANCE: Therapeutic techniques based on activation of satellite cells may block age related fiber loss and atrophy in the TA. PMID- 11020172 TI - Prevalence of reflux in 113 consecutive patients with laryngeal and voice disorders. AB - OBJECTIVES: The goal was to estimate the prevalence of laryngopharyngeal reflux (LPR) in patients with laryngeal and voice disorders. STUDY DESIGN AND SETTING: This was a prospective study of 113 unselected, new patients with laryngeal and voice disorders. Patients completed an extensive medical history form including a reflux symptom profile. A comprehensive otolaryngologic examination was performed with photographic transnasal fiberoptic laryngoscopy. Patients with both symptoms and findings of LPR (78/133, 69%) underwent ambulatory 24-hour double-probe pH monitoring. RESULTS: Seventy-three percent (57/78) of patients undergoing pH testing had abnormal studies. Thus 50% (57/113) of the entire the study population had pH-documented reflux. Of the diagnostic sub-groups studied, the highest incidence of reflux was found in patients with vocal cord neoplastic lesions (88%) and patients with muscle tension dysphonias (70%). LPR was infrequently found in patients with neuromuscular disorders. CONCLUSION: LPR occurs in at least 50% of all patients at our center with laryngeal and voice disorders at presentation. PMID- 11020173 TI - Computed tomography stage, allergy testing, and quality of life in patients with sinusitis. AB - BACKGROUND: Chronic rhinosinusitis is a common disease associated with significant morbidity. CT has been a poor predictor of the extent of symptoms among these patients. Allergic rhinitis is frequently associated with chronic rhinosinusitis. This study examines CT stage, allergic sensitivity, and sinus related symptoms. METHODS: Forty-eight patients with chronic rhinosinusitis underwent CT scanning and skin end-point titration (SET). They also completed the Rhinosinusitis Disability Index (RSDI), a measure of sinus-related disability. RESULTS: No association was noted between CT stage and severity of sinus-related symptoms. There was a significant correlation between CT stage and SET mean end point and between SET mean end point and RSDI physical scale score, as well as between significance on the RSDI emotional scale score and overall score. CONCLUSION: The degree of allergic disease is a better indicator of symptom severity and impairment in quality of life among patients with chronic rhinosinusitis than CT stage. Careful evaluation and treatment of allergic disease is critical in the treatment of chronic rhinosinusitis. PMID- 11020174 TI - Allergy and health-related quality of life. AB - OBJECTIVE: We asked patients with allergies to complete the SF-36 Health Survey, a health-related quality-of-life (QOL) measure, to determine the impact of allergy on QOL. STUDY DESIGN AND SETTING: In total, 377 adults-140 men (37%) and 237 women (63%)-seen in a tertiary care private neurotologic practice allergy clinic completed the questionnaire before beginning immunotherapy and dietary management. One hundred patients completed both initial and 1-year follow-up questionnaires. The SF-36 measures 8 health concepts ranging from physical to mental health, and scale scores range from 0 to 100. RESULTS: Initial mean scores ranged from a high of 79.1 for the physical functioning scale to a low of 47.2 for the vitality scale, lower (poorer) on all scales than norms for the general US population. Significant improvement occurred from initial to follow-up on all scales. The largest improvements were in role functioning-physical, role functioning-emotional, and social functioning. CONCLUSION: Findings suggest that allergy symptoms can affect QOL and that treatment with specific immunotherapy and/or dietary management may lead to measurable improvements. SIGNIFICANCE: The significant impact of allergy must be recognized, and treatment should be offered. The SF-36 can be used to evaluate treatment outcome. PMID- 11020175 TI - Torsional strength of the radius after osteofasciocutaneous free flap harvest with and without primary bone plating. AB - The osteocutaneous radial forearm free flap (OCRFFF) has not gained widespread popularity in mandibular reconstruction, primarily because of concerns about pathologic fracture of the weakened radius. This study examines the effectiveness of plate fixation of the radius bone after harvest of the OCRFFF as a mechanism to minimize donor-site morbidity and increase the usefulness of the OCRFFF. Matched pairs of fresh human cadaveric radius bones were used in this study. Two study groups were designed. The first group was used to define the amount of strength lost after a typical bone graft harvest. The second group was designed to demonstrate how much torsional strength was regained by the application of an orthopedic reconstruction plate. Statistically significant results were obtained for both groups. In group 1, the strength of the cut bones compared with that of the unaltered bones was significantly decreased by 82% (P = 0.016). In group 2, the cut bones reinforced with a plate were 75% stronger (P = 0.002) than the bones that were only cut. Although the radius bone is significantly weakened by the harvest of a graft, much of this strength can be regained with plate fixation of the radius. PMID- 11020176 TI - Applications of fast-setting hydroxyapatite cement: cranioplasty. AB - A variety of autogenous and synthetic materials have been used to repair cranial defects resulting from traumatic and iatrogenic causes. In theory, the ideal material should be readily available and safe. It should adequately protect the underlying central nervous system, resist cerebrospinal fluid fistula formation, and be easily contoured. One promising synthetic biomaterial that has been used for cranioplasty is hydroxyapatite cement. This biomaterial has successfully restored cranial contour in most patients in whom it has been used; however, difficulties have arisen because of the material's prolonged water solubility. When exposed to cerebrospinal fluid or blood, inadequate setting of the cement occurs, resulting in loss of its structural integrity. This problem can be alleviated with the use of fast-setting hydroxyapatite cement, which hardens 6 to 12 times faster than the traditional cement. We present, to the best of our knowledge, the first series of the use of this material in 21 patients requiring cranioplasty. The advantages and limitations of fast-setting hydroxyapatite cement will be discussed. PMID- 11020177 TI - Analysis of the human neurofibromatosis type 2 gene promoter and its expression. AB - OBJECTIVE: It is hypothesized that transcriptional regulation plays an important role for neurofibromatosis type 2 (NF2) expression in Schwann cells and other cell types. The objective of this study is the isolation and characterization of the transcriptional regulatory elements of the NF2 gene. STUDY DESIGN AND SETTING: A bacterial artificial chromosome library and a partial genomic DNA library were used to isolate the human NF2 gene; NF2 promoter-luciferase constructs were generated, and promoter activities were assayed. This study was carried out in a molecular biology laboratory. RESULTS: A bacterial artificial chromosome clone with an approximately 100-kilobase insert containing nearly the entire human NF2 gene has been isolated. An additional 5' NF2 sequence has also been cloned. Transient transfection experiments demonstrate strong promoter activity from the NF2 5' flanking DNA. CONCLUSIONS: The NF2 gene is approximately 100 kilobases long. Both positive and negative regulatory elements are present in NF2 5' flanking regions. SIGNIFICANCE: Better understanding of the NF2 gene and its regulation will improve molecular diagnostics and ultimately treatment of patients with NF2. PMID- 11020178 TI - Cochlear implant performance in senior citizens. AB - Cochlear implants are indicated for elderly patients with severe-to-profound hearing loss (sensorineural hearing loss >/=70 dB). Their use has been limited, possibly by the misconception that elderly patients will perform poorly. To document the performance of older adults (> or =65 years old), we undertook a retrospective analysis of our postlingually deafened adult patients who underwent implantation with the CLARION Multi-Strategy Cochlear Implant and underwent formal audiologic analysis (sentence recognition [Central Institute for the Deaf, CID] and monosyllabic word recognition (consonant-noun-consonant, CNC)). Both younger (n = 20; mean age = 46.9 years) and older (n = 16; mean age = 71.5 years) adults showed statistically significant increases in CID and CNC scores after cochlear implantation. No statistically significant difference could be detected in operative time, anesthesia time, length of hospitalization, or CID or CNC scores between the two age groups. We conclude that age should not be a criterion for deciding who should receive cochlear implants. PMID- 11020179 TI - Evaluation of hydroxyapatite ossicular chain prostheses. AB - Hydroxyapatite (HA) middle ear prostheses have gained popularity as an alternative to human autografts and homografts. This study reports on 3 HA prostheses types: total ossicular chain prostheses, used for grafting the stapes footplate to the tympanic membrane; partial ossicular chain prostheses, used for grafting the stapes superstructure to the tympanic membrane; and Kartush incus struts (Smith & Nephew Richards Inc), used for grafting the stapes superstructure to the undersurface of the malleus. This single-surgeon study of 33 consecutive cases revealed a statistically significant difference in mean postoperative air bone gap and airbone gap closure between incus struts (14/26 dB) or partial (22/11 dB) or total (25/10 dB) ossicular chain prostheses (t test: P<0.05). Prognostic risk factors graded by the Middle Ear Risk Index indicate a tendency for worse postoperative hearing with increasing Middle Ear Risk Index. This study supports the use of HA ossicular prostheses and, in particular, the use of the malleus for ossicular chain construction. PMID- 11020180 TI - Single-stage laryngotracheal reconstruction in children: a review of 200 cases. AB - OBJECTIVE: We reviewed our experience with pediatric single-stage laryngotracheal reconstruction (SSLTR) to identify factors that affect postoperative outcomes, including the need for reintubation and tracheostomy. STUDY DESIGN: Retrospective chart review was done. RESULTS: In total, 190 children underwent 200 SSLTRs; 29% were reintubated, and 15% required postoperative tracheostomy. Currently, 96% are decannulated. The use of anterior and posterior costal cartilage grafting, age less than 4 years, sedation for more than 48 hours, a leak pressure around the endotracheal tube at greater than 20 cm H2O, and moderate/severe tracheomalacia significantly increased the rate of reintubation. The duration of stenting did not affect outcomes. Children with anterior and posterior grafts and those with moderate or severe tracheomalacia were more likely to need a postoperative tracheostomy. CONCLUSION: SSLTR can be effective for the treatment of pediatric laryngotracheal stenosis. Diligent preoperative assessment of the patient and the patient's airway and close postoperative care are important to the success of this operation. PMID- 11020181 TI - Use of plain radiography and computed tomography to identify fish bone foreign bodies. AB - INTRODUCTION: Fish bones are the most common aerodigestive foreign bodies in adults. Radiographic studies, often ordered for diagnosis, have questionable utility. METHODS: With a laryngoscope, bones from 10 fish species were placed in a variety of positions in a human head and neck cadaver specimen. Plain films and CT scans were obtained and evaluated by blinded readers. RESULTS: Plain radiography exhibited a sensitivity and specificity of 39% and 72%. Cooking did not grossly change radio-opacity. The species of fish affected visibility in soft tissue, without correlation to the characteristic optical density of each species. Bones placed orthogonal to the film in the vallecula were most readily identified. CT scanning correctly identified 9 of 10 bones. CONCLUSIONS: Plain radiography poorly visualizes fish bone foreign bodies in soft tissue; visibility varied with fish species, location, and orientation. CT is the test of choice to radiographically diagnose fish bone impactions. PMID- 11020182 TI - Penetrating neck trauma in children: an urban hospital's experience. AB - OBJECTIVE: As the incidence of violent crime increases in our society, the rate of penetrating head and neck trauma in children also rises. The methods of management of pediatric penetrating neck wounds are addressed. METHODS: All clinical records of children younger than 18 years admitted with penetrating neck injuries between 1990 and 1997 were reviewed. The injuries were classified according to type and location of the neck wound. Demographic data, clinical presentation, diagnostic studies, and management techniques were evaluated. RESULTS: Thirty-five children aged 6 to 18 years old were evaluated for 31 missile wounds and 4 stab wounds. There were 30 boys and 5 girls. Fourteen percent of injuries were in zone 1, 60% in zone II, and 26% in zone III. Of the 33% of children with zone II penetrating neck traumas who underwent selective neck explorations, 86% had significant intraoperative findings. The mortality rates for zones I, II, and III were 60%, 29%, and 56%, respectively. The overall mortality rate was 40%. CONCLUSIONS: Penetrating neck trauma in children may lead to potentially life-threatening injuries. Selective management of penetrating head and neck injuries in children can be a safe and effective policy in an experienced trauma center. PMID- 11020183 TI - Assessment of the airway in obstructive sleep apnea syndrome with 3-dimensional airway computed tomography. AB - OBJECTIVE: The goal was to evaluate 3-dimensional airway CT for upper airway assessment in obstructive sleep apnea syndrome (OSAS). DESIGN: Airway CT was obtained and 3-dimensional airway models were constructed prospectively for 40 patients with OSAS and 10 controls. Airway dimensions were correlated with polysomnography, and comparison was made between patients with and without OSAS. RESULTS: OSAS patients had a mean respiratory distress index of 51.9 events per hour. The mean minimum cross-sectional area (XSA) in the neutral position was 67.1 mm(2). Minimum XSA decreased in both the inspiratory and expiratory phases to 16.3 mm(2) and 15.0 mm(2), respectively (P<0.001). Complete airway obstruction occurred in 1 or more phases of respiration in 28 patients. Neither airway XSA nor length of obstruction correlated with sleep apnea parameters. No statistically significant differences in airway dimensions were found between OSAS and control patients. CONCLUSIONS: Airway CT demonstrates dynamic airway obstruction in OSAS but does not correlate well with clinically important disease parameters. PMID- 11020184 TI - Common myths about electrosurgery. AB - The electrosurgery device market is lucrative and highly competitive. Several device manufacturers exist, and many creative techniques are used to differentiate products. Some device manufacturers make claims in marketing campaigns that are difficult to reconcile with the laws of physics or basic physiology. The variety of claims may be confusing to surgeons desiring to purchase new electrosurgical products. Understanding a few basic principles of electrosurgery physics can allow a surgeon to be a more informed consumer of electrosurgical products. This article discusses the basic physics of electrosurgery and then addresses several common misconceptions about electrosurgery and electrosurgical devices. PMID- 11020185 TI - Reduced-time-window sestamibi scanning for nonlocalized primary hyperparathyroidism. AB - BACKGROUND: Technetium Tc 99m sestamibi scan is the standard of care for localizing parathyroid adenomas. METHODS AND MATERIAL: We performed a retrospective, single-institution study of 111 consecutive patients with primary hyperparathyroidism who underwent standard sestamibi scan for localization of a parathyroid lesion. A revised protocol of reduced-time-window sestamibi scan was used in nonlocalized patients. The protocol was composed of single-view scans at 15, 30, 45, and 120 minutes and tomograms at 60 minutes. RESULTS: Thirty-five patients underwent surgery at our institution. Thirty-one had adenomas, and 4 had hyperplasia. Three of 31 patients did not show localization on the standard scan and underwent reduced-time-window scans, which localized 2 more adenomas. These were confirmed at the time of surgery. CONCLUSION: We suggest that all patients with a high degree of suspicion for a parathyroid adenoma, which is not visualized by a standard scan, undergo a reduced-time-window scan. PMID- 11020186 TI - Tumors and tumorous masses presenting as temporomandibular joint syndrome. AB - OBJECTIVE: Neoplasms of the temporomandibular joint (TMJ) usually mimic common causes of TMJ syndrome, leading to delay in diagnosis. To increase awareness of TMJ neoplasms and establish guidelines for early intervention, we performed a retrospective analysis of a series of patients with neoplasms of the TMJ. STUDY DESIGN AND SETTING: A retrospective review of the records of patients with neoplasms of the TMJ from 1990 to 1997 was done. RESULTS: Six patients were identified. The neoplasms included benign and malignant neoplasms. The time from initial presentation to final diagnosis was in most cases prolonged, ranging from 3 months to 8 years. Patients typically showed advanced lesions radiographically. All patients required surgical extirpation. Postoperative radiation therapy was used for malignant lesions. Patient outcomes were delineated. CONCLUSIONS AND SIGNIFICANCE: Neoplasms, both benign and malignant, of the TMJ are rare but represent a challenging diagnostic problem. In patients with 1 month or more of recalcitrant TMJ swelling or pain, radiographic imaging may be considered to rule out these rare neoplasms. This may lead to earlier intervention and improved outcome. PMID- 11020187 TI - Endoscopic surgery for a parathyroid functioning adenoma resection with the neck region-lifting method. AB - Recently, endoscopic surgery has been applied to cervical exploration. We have developed new techniques for endoscopic neck surgery. We reported on a 53-year old Japanese man with functioning parathyroid adenoma resected by endoscopic surgery with a neck region-lifting method. A 10-mm midline trocar for the endoscope and two 5-mm lateral trocars were inserted from the anterior chest wall to avoid neck scars. There were no intraoperative complications. The incisions were completely covered by the patient's undergarments. PMID- 11020188 TI - New approach for cochlear implantation: cochleostomy through the middle fossa. AB - The middle fossa approach was used in 11 patients with profound bilateral hearing loss for insertion of a cochlear implant. Fibroadhesive otitis media (n = 1), bilateral cavity radical mastoidectomy (n = 1), autoimmune inner ear disease (n = 2), previous cranial trauma (n = 1), genetic prelingual deafness (n = 5), and otosclerosis (n = 1) were the causes of deafness. A cochleostomy was performed on the most superficial part of the basal turn, and the electrode array was inserted up to the cochlear apex. Speech perception tests (1-9 months after cochlear implant activation) yielded better results in these patients compared with a homogeneous group of postlingually deaf patients operated on through the traditional transmastoid route. Insertion of the implant through the middle fossa cochleostomy furnishes the possibility of stimulating areas of the cochlea (ie, the middle and apical turns) where a greater survival rate of spiral ganglion cells is known to occur, with improvement of information regarding the formants relevant for speech perception. PMID- 11020189 TI - The nature of juvenile nasopharyngeal angiofibroma. AB - OBJECTIVE: The goal of this study was to investigate the nature of juvenile nasopharyngeal angiofibroma. STUDY DESIGN: Twenty-five excised tumors were studied morphologically to confirm the histopathologic features of the tumor. METHODS: Paraffin sections of all tumor specimens were stained with hematoxylin eosin for elastic fiber, polyclonal antibody to factor VIII-related antigen and monoclonal antibodies to smooth muscle actin, estrogen receptor, and progesterone receptor. A quantitative assessment of the vascular component changes was also performed in 14 cases. Ultrathin sections of 5 tumors were studied electromicroscopically. Particular attention was paid to the progressive changes of both vascular and fibrous components as well as their mutual relationship. RESULTS: (1) Beneath the endothelium of the vascular component, there was a thin layer of spindle cells; these cells had a positive reaction for smooth muscle actin, thus proving to be pericytes. (2) The actively proliferating site of the tumor was usually at the peripheral portion, with abundant vascular element lined by plump endothelial cells. (3) At the central and base locations, the growth of tumor tissue usually appeared to be in a relatively resting state. The vascular component decreased in amount as the extracellular collagenous fibers increased. CONCLUSIONS: Because of the bicellular mixed composition of the vascular component and the tendency of synchronous maturation of both vascular and fibrous elements, it is suggested that juvenile nasopharyngeal angiofibroma is similar to most hemangiomas by nature and is likely a vascular hamartoma or a pathologic and reactive proliferation of vessels. PMID- 11020190 TI - Pitfalls in the surgical management of cancer of the larynx and hypopharynx. AB - This article is based on findings in 180 patients with cancer of the larynx and hypopharynx. Positive margins were found in 12.2% (22/180) of all tumors, 12.5% (8/64) of supraglottic tumors, 16% (4/25) of transglottic tumors, 5.3% (2/38) of glottic tumors, 11.1% (2/18) of pyriform fossa tumors, 50% (3/6) of posterior pharyngeal wall tumors, and 10.6% (3/29) of postcricoid tumors. The relationship between the inadequate surgical margins of the specimens, tumor spread, preoperative CT findings, and surgical procedures performed have been correlated. PMID- 11020191 TI - Comparative external versus endoscopic dacryocystorhinostomy: results in 115 patients (130 eyes). AB - BACKGROUND: Epiphora is an annoying symptom, embarrassing the patient both socially and functionally. The two widely accepted treatment modalities of epiphora resulting from obstruction of the nasolacrimal ductus are external and endoscopic dacryocystorhinostomy (DCR). OBJECTIVE: The aim was to compare the results, operative time, and complications of external and videoendoscopic endonasal DCR performed between December 1994 and December 1998. METHODS: In group 1, conventional primary external DCR with or without silicone tube intubation was performed in 79 patients (66 women and 13 men) with unilateral dacryocystitis. In group 2, endoscopic primary endonasal DCR with hammer-chisel removal of bone located over the lacrimal sac was performed in 51 eyes of 36 patients, 33 women and 3 men (15 bilateral procedures). RESULTS: The follow-up period was 6 to 48 months (mean 25 months) after surgery. The age range was from 4 to 76 years (mean 38.5 years). The success rates of external and endoscopic hammer-chisel DCR were found to be 89.8% and 88.2%, respectively. A lower complication rate was observed in the endoscopic group, with minimal morbidity and shorter operative time compared with the external approach. CONCLUSIONS: Hammer-chisel endoscopic DCR is practical, less traumatic, less time-consuming, and cosmetically more convenient than the external approach. The success rate of the endoscopic DCR procedure is comparable with that of traditional external DCR, and it also allows simultaneous correction of any intranasal pathology. PMID- 11020192 TI - Inlay butterfly cartilage tympanoplasty (Eavey technique) modified for adults. AB - INTRODUCTION: Popular myringoplasty techniques include either an underlay or an onlay approach using tissues such as temporalis fascia or perichondrium as grafts. In 1998, Eavey described an inlay tympanoplasty technique in children using a cartilage graft which resembled butterfly wings. This technique was effective for closure of tympanic membrane perforations, while being more rapid for the surgeon and more comfortable for the patient. The aim of this study was to describe our results with modification of this technique in adults under local anesthesia. METHODS AND MATERIAL: Cartilage butterfly inlay tympanoplasty was carried out in 20 adult patients to close small-to-medium tympanic membrane perforations over a period of 11 months. Compared with the original description, 3 modifications are noted: (1) adult patients, (2) local anesthesia, and (3) no skin graft over the cartilage. We evaluated the success rate and postoperative hearing results. RESULTS: Two patients were not evaluated in terms of hearing gain because they had profound preoperative hearing loss. Fourteen patients had a postoperative 4 pure-tone average between 0 and 9 dB, 3 patients between 10 and 20 dB, and 1 patient greater than 20 dB. The graft "take rate" was 90% (18/20) at the end of the follow-up period (mean follow-up 7.25 months). Seventeen of 18 successful cases had cartilage grafts that were intact and dry at the last visit. CONCLUSIONS: The tympanoplasty technique described by Eavey is effective to close tympanic membrane perforation in adults as well as children. The technique modifications introduced here (abandonment of the skin graft and local anesthesia) reduced surgery time and postoperative morbidity. Postoperative results compared with those of the original technique were good. PMID- 11020193 TI - Tracheotomy complications: a retrospective study of 1130 cases. AB - BACKGROUND: Tracheotomy is one of the most frequently performed surgical procedures in the critically ill patient. It is frequently performed as an elective therapeutic procedure and only rarely as an emergency procedure. Complications occur in 5% to 40% of tracheotomies depending on study design, patient follow-up, and the definition of the different complications. The mortality rate of tracheotomy is less than 2%. Numerous studies demonstrate a greater complication and mortality rate in emergency situations, in severely ill patients, and in small children. METHODS: A retrospective study of 1130 consecutive tracheotomies performed during 1 decade (January 1987 through December 1996) is presented. We studied the indications for surgery, the major complications of tracheotomy, and their treatment and outcome. We also noted the overall mortality rate and the specific complications that led to these deaths. RESULTS: In total, 1130 tracheotomies were performed. Major complications occurred in 49 of the cases, and 8 deaths were directly attributed to the tracheotomy. The most common complication was tracheal stenosis, which occurred in 21 cases. Hemorrhage was the second most common complication, which occurred in 9 cases. CONCLUSION: This is one of the largest series of consecutive tracheotomies compiled. We found a relatively low overall complication and mortality rate compared with other large series. Tracheal stenosis was the most common complication in contrast to other series. Our opinion is that this may reflect tracheal damage originally caused by prolonged intubation before the tracheotomy. We believe that all other complications of tracheotomy may be prevented or minimized by careful surgical technique and postoperative tracheotomy care. PMID- 11020194 TI - Influence of height on the spatial orientation and equilibrium of the body. AB - The sway of the center of gravity for 30 seconds with a stabilometer was examined in 30 volunteers under 3 visual conditions (eyes open with and without fixation and eyes closed) at heights of 0, 1, and 2 m and under 2 visual conditions (eyes open with fixation and eyes closed) at a height of 10 m 22 cm. Eight of the subjects had acrophobia. The sway worsened at 10 m 22 cm but showed no change at 1 or 2 m. The acrophobic group became clearly worse at 10 m 22 cm. In another group of 12 volunteers, the subjects were guided to the roof with the eyes closed, and sway was measured. Sway was also measured with the eyes uncovered and open and again with the eyes closed. The first measurement with the eyes closed showed worse sway, and the second with the eyes open was better, except in 2 subjects who were acrophobic. PMID- 11020195 TI - Diagnostic value of Epstein-Barr viral serology in nasopharyngeal carcinoma. AB - Although Epstein-Barr virus (EBV) IgA serology has been established as an effective marker for nasopharyngeal carcinoma (NPC), it remains unclear how useful or cost-effective it is as a screening test. This article is aimed at establishing how these tests could be used most effectively in the diagnosis of NPC in an otolaryngology outpatient clinic. A total of 111 consecutive patients with NPC and an equal number of control subjects were studied. Forty-seven patients with NPC had early (AJCC stages 1 and 2) and 64 had advanced (stages 3 and 4) disease. A positive early antigen (EA) serology result was found in 81.2% of NPC patients and in none of the controls. Negative EA and viral capsid antigen (VCA) serology results were present in 2.7% of NPC patients and in 46.8% of controls. Negative EA and positive VCA serology results were found in 30.0% of NPC patients with early disease, 7.8% of NPC patients with advanced disease, and 53.2% of controls. Given its high specificity, serology for EA is recommended as a clinically useful screening test. Serology for VCA, although highly sensitive, has an unacceptably high false-positive rate, and its cost-effectiveness as a universal screening test is questionable. PMID- 11020196 TI - Amplification of herpes simplex virus type 1 DNA in human geniculate ganglia from formalin-fixed, nonembedded temporal bones. AB - Polymerase chain reaction (PCR) has provided new insights in molecular biology. Recently, some studies have been focused on temporal bone pathology, with amplification of DNA from fixed sections of celloidin-embedded bones. The purpose of our study was to elucidate the utility of PCR in detection of minor concentrations of DNA from nonoptimal stored samples. We obtained geniculate ganglia from 30 temporal bones preserved in formalin for a long time, without any process of embedding. By performing a nested PCR assay, we detected herpes simplex virus type 1 DNA in 13 of 30 ganglia (43%). We conclude therefore that study of temporal bones stored under poor conditions by PCR is possible, although there are some limitations when compared with fresh or optimally archived samples. PMID- 11020197 TI - Extended lateral rhinotomy incision for total maxillectomy. AB - An extended lateral rhinotomy incision for total maxillectomy, which avoids postoperative lower eyelid complications, is described. Advantages of the extended lateral rhinotomy incision for total maxillectomy are discussed in comparison with the classic Weber-Fergusson incision. PMID- 11020198 TI - Round window obliteration by osteoma of the middle ear. PMID- 11020200 TI - Hemangioma of the ethmoid sinus. PMID- 11020199 TI - Salvage of an infected hydroxyapatite cement cranioplasty with preservation of the implant material. PMID- 11020201 TI - Sarcoidosis presenting as recurrent nasal polyps. PMID- 11020202 TI - Dehiscence of the tympanic segment of the facial nerve. PMID- 11020203 TI - Choristoma of the nasopharynx. PMID- 11020204 TI - Epistaxis: new approach. PMID- 11020205 TI - Secondary intention healing of exposed scalp and forehead bone. PMID- 11020206 TI - Secondary intention healing of exposed scalp and forehead bone. PMID- 11020207 TI - In search of a hammer. PMID- 11020208 TI - In search of a hammer PMID- 11020209 TI - The hypocretins: excitatory neuromodulatory peptides for multiple homeostatic systems, including sleep and feeding. AB - The hypocretins are two neuropeptides of related sequence that are produced from a common precursor whose expression is restricted to 1, 100 large neurons of the rat dorsal-lateral hypothalamus. The hypocretins have been detected immunohistochemically in secretory vesicles at synapses of fibers that project to areas within the posterior hypothalamus that are implicated in feeding behaviors and hormone secretion and diverse targets in other brain regions and in the spinal cord, including several areas implicated in cardiovascular function and sleep-wake regulation. The hypocretin-producing cells have receptors for leptin and receive input from arcuate neuropeptide Y neurons. The peptides are excitatory when applied to cultured hypothalamic, cortical, or spinal cord neurons. Two G protein-coupled receptors for the hypocretins have been identified, and these have different distributions within the CNS and differential affinities for the two hypocretins. Administration of the hypocretins stimulates food intake; affects blood pressure, hormone secretion, and locomotor activity; and increases wakefulness while suppressing REM sleep. The hypocretin mRNA accumulates during food deprivation. An inactivating insertion into the hypocretin receptor 2 gene in dogs results in narcolepsy. Mice whose hypocretin gene has been inactivated exhibit a narcolepsy-like phenotype. Human patients with narcolepsy have greatly reduced levels of hypocretin peptides in their cerebral spinal fluid. One aspect of hypocretin activity is the direct excitation of noradrenergic neurons in the locus coeruleus to prevent entry into REM sleep. These peptides appear to be part of a complex circuit that integrates aspects of energy metabolism, cardiovascular function, hormone homeostasis, and sleep-wake behaviors. PMID- 11020210 TI - Artificial gravity as a countermeasure in long-duration space flight. AB - Long-duration exposure to weightlessness results in bone demineralization, muscle atrophy, cardiovascular deconditioning, altered sensory-motor control, and central nervous system reorganizations. Exercise countermeasures and body loading methods so far employed have failed to prevent these changes. A human mission to Mars might last 2 or 3 years and without effective countermeasures could result in dangerous levels of bone and muscle loss. Artificial gravity generated by rotation of an entire space vehicle or of an inner chamber could be used to prevent structural changes. Some of the physical characteristics of rotating environments are outlined along with their implications for human performance. Artificial gravity is the centripetal force generated in a rotating vehicle and is proportional to the product of the square of angular velocity and the radius of rotation. Thus, for a particular g-level, there is a tradeoff between velocity of rotation and radius. Increased radius is vastly more expensive to achieve than velocity, so it is important to know the highest rotation rates to which humans can adapt. Early studies suggested that 3 rpm might be the upper limit because movement control and orientation were disrupted at higher velocities and motion sickness and chronic fatigue were persistent problems. Recent studies, however, are showing that, if the terminal velocity is achieved over a series of gradual steps and many body movements are made at each dwell velocity, then full adaptation of head, arm, and leg movements is possible. Rotation rates as high as 7.5-10 rpm are likely feasible. An important feature of the new studies is that they provide compelling evidence that equilibrium point theories of movement control are inadequate. The central principles of equilibrium point theories lead to the equifinality prediction, which is violated by movements made in rotating reference frames. PMID- 11020211 TI - Long-term potentiation in the presence of NMDA receptor antagonist arylalkylamine spider toxins. AB - The role of the NMDA receptor (NMDAR) in long-term potentiation (LTP) is now well established. All potent NMDAR antagonists known to date inhibit the induction of LTP at the Schaffer collateral-CA1 pyramidal cell synapse in rat hippocampus, regardless of their site and mechanism of action. Arylalkylamine toxins are noncompetitive NMDAR antagonists in the mammalian central nervous system (CNS). The synthetic toxins argiotoxin-636 (Arg-636), Joro spider toxin (JSTX-3), alpha agatoxin-489 and -505 (Agel-489 and Agel-505) and philanthotoxin-433 (delta-PhTX) were found in the present study to have no effect on the induction of LTP in the Schaffer collateral-CA1 pyramidal cell pathway in rat hippocampal slices maintained in vitro. Arylalkylamine toxins represent a class of potent NMDAR antagonists that fail to affect hippocampal LTP, and thus provide novel structural leads for the development of NMDAR antagonists that do not impair cognition. PMID- 11020212 TI - Expression and role of sulfoglucuronyl (HNK-1) carbohydrate and its binding protein SBP-1 in developing rat cerebral cortex. AB - Developmental expression of sulfoglucuronyl carbohydrate (SGC) and its binding protein, SBP-1 was studied in the rat cerebral cortex to understand their function. Between embryonic day (ED) 14-19, SBP-1 was strongly expressed in neurons of the ventricular zone and migrating neurons throughout the cortex. SBP 1 declined at birth and by postnatal day (PD) 3 only the latest arriving neurons in the most superficial segment of the cortical plate expressed SBP-1. Between ED 14-16, SGC was expressed in a thin row of glial cells near the ventricles and on their radial processes. Between ED 16-PD 3, SGC was not in neuronal cell soma, but was in neuronal plasma membranes and processes surrounding the neuronal perikarya. The expression of SGC declined similar to SBP-1 and both of them disappeared by PD 7. The expression of SBP-1 and SGC was chronologically coordinated with neuronal migration. SBP-1 was specifically expressed in immature neuronal nuclei and plasma membranes. SBP-1 and SGC were colocalized and were available for interaction with each other on neuronal cell membranes and processes. This was confirmed with isolated neurons in culture. As in vivo, the expression of SBP-1 in neurons declined with time in culture. The dissociated cortical neurons when plated on SBP-1 as a substratum produced extensive neuritic outgrowth. HNK-1, anti-SBP-1 antibodies and sulfoglucuronyl glycolipid, SGGL specifically and severely reduced neurite outgrowth. SBP-1-SGC interactions provide a potential mechanism for guidance and cell signaling, in the processes of neuronal migration and terminal differentiation. PMID- 11020213 TI - Molecular cloning of rat alpha1,3-fucosyltransferase IX (Fuc-TIX) and comparison of the expression of Fuc-TIV and Fuc-TIX genes during rat postnatal cerebellum development. AB - Fucosylated glycoconjugates play an essential role in central nervous system development, but the regulation of expression of these molecules is not well understood. The final biosynthetic step for a major group of cerebellar fucosylated glycoconjugates (those bearing the developmentally regulated epitope 3-fucosyl-N-acetyllactosamine, CD15, and related fucosylated epitopes) is catalyzed by an alpha-1,3-fucosyltransferase (FucT). The major FucT activity in postnatal rat cerebellum has a specificity consistent with that encoded by either a Fuc-TIV- or Fuc-TIX-like gene, and thus the expression of these genes was investigated during postnatal rat cerebellar development. A rFuc-TIX cDNA was cloned and a comparison of its enzymatic activity with rFuc-TIV revealed similar results on oligosaccharides, but strikingly higher activity on lipid acceptors, suggesting a greater role for rFuc-TIX than rFuc-TIV in the synthesis of CD15 glycolipids. rFuc-TIX mRNA levels were much higher than those of rFuc-TIV in neonatal cerebellum. Whereas rFuc-TIX mRNA levels remained relatively constant, rFuc-TIV mRNA levels declined during postnatal cerebellar development. In situ hybridization of postnatal rat cerebella also revealed different patterns of expression for these two genes. The rFuc-TIV gene was expressed predominantly in Purkinje cells and the deep cerebellar nuclei throughout postnatal development, but was expressed in granule neurons only in the neonatal, and not the adult, rat. In contrast, the rFuc-TIX gene was expressed in cells in the granule cell layers in both neonatal and in the adult rat. The potential implications of the different enzymatic activities and cell localization of rFuc-TIV and rFuc-TIX expression for the regulation of fucosylated glycoconjugates during cerebellar development are discussed. PMID- 11020214 TI - PtdIns 4-kinasebeta and neuronal calcium sensor-1 co-localize but may not directly associate in mammalian neurons. AB - It was recently demonstrated that the yeast homologue of phosphatidylinositol 4 kinasebeta PIK1 is directly associated with frq1, the yeast homologue of mammalian neuronal calcium sensor-1 (NCS-1) (Hendricks et al., [1999] Nat. Cell Biol. 1:234- 241). This was a novel finding and suggests that a calcium binding protein activates and regulates PtdIns 4-kinasebeta. This finding had not been shown in mammalian cells and both PtdIns 4-kinasebeta and NCS-1 have been shown to have important roles in the regulation of exocytotic release associated with neurotransmission. The aims of this study were to determine if PtdIns 4 kinasebeta and NCS-1 directly associate in mammalian neural tissues. We show that the immunostaining pattern for PtdIns 4-kinasebeta and NCS-1 is co-localized throughout the neurites of newborn cultured dorsal root ganglia (DRG) neurons but not in E13 DRG neurons. We then provide biochemical evidence that PtdIns 4 kinasebeta may not be in physical association with NCS-1 in mammalian nervous tissue unlike that previously reported in yeast. PMID- 11020215 TI - Palmitoylation of the p75 neurotrophin receptor has no effect on its interaction with TrkA or on TrkA-mediated down-regulation of cell adhesion molecules. AB - The short- and long-term effects of nerve growth factor (NGF) were studied on fibroblast cell lines stably expressing both TrkA and either wild-type p75 or a mutant that lacks the palmitoylation site of p75. The lack of palmitoylation had no effect on the ability of p75 to enhance the short-term NGF-induced tyrosine phosphorylation of TrkA over a wide range of NGF concentrations. Long-term treatment of the cell lines with NGF led to loss of cell adhesion to the culture dishes that increased with increasing concentrations of NGF and increased expression of TrkA. Treatment of the cell lines with mutant NGFs that bound selectively to TrkA or p75 alone revealed that cell detachment was mediated solely through TrkA. Increased cell detachment correlated with a decrease in the expression levels of fibronectin and cadherin, cell surface molecules involved in cell adhesion. The loss of cell adhesion with the cell line expressing the palmitoylation-deficient p75 were identical to those expressing wild type, as was anticipated from the lack of involvement of p75 in this process. PMID- 11020216 TI - Antisense palmitoyl protein thioesterase 1 (PPT1) treatment inhibits PPT1 activity and increases cell death in LA-N-5 neuroblastoma cells. AB - Infantile neuronal ceroid lipofuscinosis (INCL) is a childhood neurodegenerative disease caused by the selective death of cortical neurons and retinal degeneration, as the result of a palmitoyl protein thioesterase 1 (PPT1) deficiency. Recently, we showed that overexpression of PPT1 protects LA-N-5 human neuroblastoma cells against apoptotic death (Cho and Dawson [2000a] J. Neurochem. 74:1478-1488) and we now show that inhibition of PPT1 increases the susceptibility of these cells to apoptotic cell death. Transient transfection of LA-N-5 neuroblastoma cells with PPT1-FLAG resulted in a strong expression of PPT FLAG-tagged protein as evidenced by Western blot analysis and immunofluorescence. Co-transfection of a reverse-oriented (antisense) PPT1 (AS-PPT1) decreased the expression of PPT-FLAG to almost zero, reduced PPT1 enzyme activity (as measured by an in vitro assay) and increased the susceptibility to apoptosis induced by C(2) ceramide. Similarly, inhibition of PPT1 with a synthetic inhibitor (AcG palmitoyl diaminoproprionate-VKIKK) (DAP1) (100 microM) increased the susceptibility of the cells to apoptosis induced by either C(2)-ceramide or etoposide, a common chemotherapeutic agent used in the treatment of neuroblastoma. Cells stably overexpressing PPT1 were resistant to apoptosis induced by DAP1 suggesting that the inhibitor has a specific action and confirming that low levels of protein palmitoylation block the death pathway. Drugs that raise the level of protein palmitoylation are pro-apoptotic and PPT1 inhibition may enhance the killing efficacy of chemotherapeutic agents used to kill neuroblastoma-derived cells. PMID- 11020217 TI - Fibroblast growth factor 2 (FGF2) and FGF receptor expression in an experimental demyelinating disease with extensive remyelination. AB - Fibroblast growth factor 2 (FGF2) is an excellent candidate to regulate remyelination based on its proposed actions in oligodendrocyte lineage cell development in conjunction with its involvement in CNS regeneration. To assess the potential for FGF2 to play a role in remyelination, we examined the expression pattern of FGF2 and FGF receptors (FGFRs) in an experimental demyelinating disease with extensive remyelination. Adult mice were intracranially injected with murine hepatitis virus strain A-59 (MHV-A59) to induce focally demyelinated spinal cord lesions that spontaneously remyelinate, with corresponding recovery of motor function. Using kinetic RT-PCR analysis of spinal cord RNA, we found significantly increased levels of FGF2 mRNA transcripts, which peaked during the initial stage of remyelination. Analysis of tissue sections demonstrated that increased levels of FGF2 mRNA and protein were localized within demyelinated regions of white matter, including high FGF2 expression associated with astrocytes. The expression of corresponding FGF receptors was significantly increased in lesion areas during the initial stage of remyelination. In normal and lesioned white matter, oligodendrocyte lineage cells, including progenitors and mature cells, were found to express multiple FGFR types (FGFR1, FGFR2, and/or FGFR3). In addition, in lesion areas, astrocytes expressed FGFR1, FGFR2, and FGFR3. These findings indicate that, during remyelination, FGF2 may play a role in directly regulating oligodendrocyte lineage cell responses and may also act through paracrine or autocrine effects on astrocytes, which are known to synthesize other growth factors and immunoregulatory molecules that influence oligodendrocyte lineage cells. PMID- 11020218 TI - Group I and group III metabotropic glutamate receptor subtypes provide enhanced neuroprotection. AB - Neuroprotection by the metabotropic glutamate receptor (mGluR) system has been linked to the modulation of both the free radical nitric oxide (NO) and programmed cell death (PCD). Because the cellular mechanisms that ultimately determine neuronal PCD rely upon the independent pathways of genomic DNA degradation, externalization of membrane phosphatidylserine (PS) residues, and the activation of associated cysteine proteases, we investigated the ability of the individual mGluR subtypes to modulate the distinct pathways of NO-induced PCD in primary rat hippocampal neurons. Membrane PS residue externalization occurred within the initial 3 hr after exposure to the NO donors (300 microM SNP or 300 microM NOC-9), preceded genomic DNA fragmentation, and was present in 80 +/- 2% of the neurons within a 24-hr period. NO exposure also led to the rapid induction of both caspase 1-like and caspase 3-like activities that were determined to be necessary, at least in part, for the generation of NO-induced genomic DNA degradation, but distinct from the detrimental effects of intracellular acidification. Yet, only caspase 1-like activity was necessary for the modulation of PS residue externalization. Activation of group I mGluR subtypes utilized an effective, "upstream" mechanism for the inhibition of cysteine protease activity that offered an enhanced level of neuroprotection through both the preservation of genomic DNA integrity and the maintenance of PS membrane asymmetry. Group II and Group III mGluR subtypes maintained DNA integrity and group III mGluR subtypes additionally prevented PS residue externalization through mechanisms that were targeted below the level of caspase activation. Our work elucidates the independent nature of the mGluR subtypes to not only provide discrete levels of protection against neuronal PCD, but also offer robust therapeutic strategies for neurodegenerative disease. PMID- 11020219 TI - Kinetics of facial motoneuron loss following facial nerve transection in severe combined immunodeficient mice. AB - We have recently shown that cells of the acquired immune system are crucial components of motoneuron survival after injury (Serpe et al. [1999] J. Neurosci. 19:RC7). The goal of the present study was to determine the kinetics of facial motoneuron (FMN) loss in wild-type, scid, and reconstituted scid mice after a right facial nerve axotomy at the stylomastoid foramen. Scid mice showed a significant decrease in FMN survival at all weekly postoperative (wpo) times. One, two, four, and ten wpo, ipsilateral FMN survival in scid mice was 90% +/- 1.8%, 84% +/- 1.3%, 52% +/- 3.7%, and 45% +/- 2.5%, respectively, of the contralateral, unoperated side. In contrast, FMN loss after axotomy in wild-type and reconstituted scid mice was not observed until 4 wpo (86% +/- 2.5% and 83% +/ 3.5%, respectively) relative to the contralateral, unoperated side. However, the levels of FMN in both wild-type and reconstituted scid mice were significantly higher than those in the nonreconstituted scid at 4 wpo. By 10 wpo, FMN survival in both wild-type and reconstituted scid mice had continued to decline significantly (60% +/- 2.1% and 58% +/- 3.1%, respectively) relative to the contralateral, unoperated side but were still significantly higher than that of the nonreconstituted scid at 10 wpo. Several important controls were also added to this study. Because the scid mutation is present in all cells (although it specifically results in a loss of V(D)J recombination mechanisms, we wanted to rule out the actual DNA mutation as causal in FMN loss). To accomplish this, we used the recombinase-activating gene-2 knockout (RAG-2 KO) mouse model, in which the RAG-2 has been disrupted and prevents maturation of T and B cells. As with the scid model, there was a significant loss of FMN at 4 wpo in the RAG-2 KO that could be reversed with whole splenocyte reconstitution. We also compared FMN numbers in nonaxotomized facial nuclei from both scid and RAG-2 KO mice relative to wild-type controls. No differences in normal numbers of FMN were found in either the mutation or the gene knockout model. The ability of T and B lymphocytes to rescue FMN from cell death after peripheral nerve injury supports the hypothesis that cells of the acquired immune system produce neurotrophic factors or neurocytokines to support neuronal survival until target reconnection occurs. PMID- 11020220 TI - Effect of acute hyperglycemia on visual cortical activation as measured by functional MRI. AB - To determine whether acute hyperglycemia changes the hyperemic response to functional activation of brain, the area and magnitude of the activation were measured in healthy volunteers maintained at euglycemia and then at hyperglycemia using the hyperglycemic clamp technique. Activation of the visual cortex (8-16 Hz) was assessed by functional MRI with blood oxygenation level dependent (BOLD) contrast using a 4 Tesla magnet and a multi-slice echo-planar imaging sequence (TE = 30 msec, TR = 1.5 sec). At euglycemia (4.8 +/- 0.2 mM, mean +/- SEM, n = 6), the number of activated pixels in the occipital lobe was 79 +/- 10 and the intensity of activation was 4.5 +/- 0.5%. During hyperglycemia (plasma glucose 300% of control), the number of activated pixels was 90 +/- 20% of control and the BOLD activation was 3.5 +/- 0.3%, respectively. The change in BOLD signal was below 0.2%/mM plasma glucose. This study demonstrates that acute hyperglycemia is without substantial effect on the size and intensity of activation of the occipital cortex. The results further suggest that fluctuations in blood glucose within the physiologic range are without effect on the functional activation of the cerebral cortex measured by BOLD fMRI. PMID- 11020221 TI - Thiamine deficiency results in metabolic acidosis and energy failure in cerebellar granule cells: an in vitro model for the study of cell death mechanisms in Wernicke's encephalopathy. AB - Thiamine deficiency (TD) in both humans and experimental animals results in severe compromise of mitochondrial function and leads to selective neuronal cell death in diencephalic and cerebellar structures. To examine further the influence of TD on neuronal survival in relation to metabolic changes, primary cultures of rat cerebellar granule cells were exposed to thiamine-deficient medium for up to 7 days in the absence or presence of the central thiamine antagonist pyrithiamine (Py). Exposure of cells for 7 days to thiamine-deficient medium alone resulted in no detectable cell death. On the other hand, 50 microM Py treatment led to reductions of thiamine phosphate esters, decreased activities of the thiamine dependent enzymes alpha-ketoglutarate dehydrogenase and transketolase, a twofold increase in lactate release (P < 0.001), a lowering of pH, and significant (58%, P < 0.001) cell death. DNA fragmentation studies did not reveal evidence of apoptotic cell death. Addition of 50 microM alpha-tocopherol (vitamin E) or 100 microM of butylated hydroxyanisole (BHA) to Py-treated cells resulted in significant neuroprotection. On the other hand, addition of 10 microM MK-801, an NMDA receptor antagonist, was not neuroprotective. These results suggest that reactive oxygen species (ROS) play a major role in thiamine deficiency-induced neuronal cell death. Insofar as this experimental model recapitulates the metabolic and mitochondrial changes characteristic of thiamine deficiency in the intact animal, it might be useful in the elucidation of mechanisms involved in the neuronal cell death cascade resulting from thiamine deficiency. PMID- 11020222 TI - Amyloidogenicity and neurotoxicity of peptides corresponding to the helical regions of PrP(C). AB - An alpha-helical to beta-sheet conformational change in the prion protein, PrP(C), is believed to be causative in transmissible spongiform encephalopathies. Recent nuclear magnetic resonance structures of PrP(C) have identified three helical regions in the normal full-length protein. We have synthesised peptides corresponding to these helical regions (PrP144-154, helical region one; PrP178 193, helical region two; and PrP198-218, helical region three). Circular dichroism results show that the peptide corresponding to helical region one is unstructured, while peptides corresponding to the second and third helical regions have a high propensity to form beta-sheet structure in a pH-dependent manner in aqueous solutions. Peptides corresponding to the second helical region, PrP180-193 and PrP178-193, are the only ones that form amyloid by electron microscopy and congo red birefringence. PrP178-193 and the amyloidogenic Alzheimer's disease Abeta25-25 peptide were found to promote Cu (II)-induced lipid peroxidation and cytotoxicity in primary neuronal cultures, while PrP144 154, PrP198-218 and the nonamyloidogenic Abeta1-28 had no effect on Cu (II) toxicity. There was no increase in toxicity induced by PrP178-193 in cultures treated with Fe (II) or hydrogen peroxide, indicating a preferential modulatory effect on Cu (II) toxicity by PrP178-193. The data suggest that the PrP178-193 peptide has both structural and bioactive properties in common with Abeta25-35 and that the second putative helical region of PrP could be involved in modulation of Cu (II)-mediated toxicity in neurons during prion disease. PMID- 11020223 TI - Defective ubiquitination of cerebral proteins in Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by the presence of neurofibrillary tangles (NFT), senile plaques, and cerebrovascular deposits of amyloid-beta. Ubiquitin has also been shown to be present in some of the inclusions characteristic of this disease. To obtain further insight into the role played by the ubiquitin pathway in AD, we investigated the capacity of postmortem samples of cerebral cortex from normal and AD patients to form high-molecular-weight ubiquitin-protein conjugates. Activity of the ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzymes (E2) involved in the ubiquitin pathway was also determined. In normal samples, the amount of high-molecular-weight ubiquitin protein conjugates (HMW-UbPC) in cytosol increased with incubation time, whereas, in samples of AD cases, these were almost undetectable. The addition of an adult rat fraction, enriched in ubiquitinating enzymes, restored the capacity of AD brain cytosolic fraction to form conjugates. The trypsin-like proteolytic activity of the 26S proteasome was found to be decreased in AD cytosol brain. Assay of the activity of E1 and E2 by thiol-ester formation revealed a significant decrease in AD samples. Moreover, Western blotting using a specific antibody against E1 showed a dramatic drop of this enzyme in the cytosolic fraction, whereas normal levels were found in the particulate fraction, suggesting a possible delocalization of the enzyme. Our results suggest that a failure in the ubiquitination enzymatic system in brain cytosol may contribute to fibrillar pathology in AD. PMID- 11020224 TI - Notch-1 activation by familial Alzheimer's disease (FAD)-linked mutant forms of presenilin-1. AB - We prepared a cleavage site-directed antibody against Notch-1, that specifically recognized the cleaved Notch-1 intracellular domain (NICD). To assess Notch-1 processing and its nuclear localization in familial Alzheimer's disease (FAD) linked presenilin-1 (PS-1) mutants, we overexpressed wild type, M146V, A246E, C410Y, or deltaE9 PS-1 mutant with a membrane-bound Notch-1 in a PS-1-deficient cell line. On Western blot and immunocytochemical analyses using the NICD specific antibody, M146V and A246E mutants showed the comparable levels of Notch 1 processing and nuclear localizing activities to wild type PS-1 whereas C410Y and deltaE9 mutants failed to show these activities. These results suggest that the loss or partial loss of PS-1 activities in Notch-1 proteolysis and its nuclear translocation may be irrelevant for the neuropathology of Alzheimer's disease. PMID- 11020225 TI - Observation of the ultrastructure and synaptic relationships of angiotensin II like immunoreactive neurons in the rat area postrema. AB - The ultrastructure and synaptic relationships of the angiotensin II-containing neurons in the area postrema of the rat were studied by immunocytochemistry using the avidin-biotin-complex-DAB method, and also using silver-gold intensification following the DAB reaction. At the light microscopic level, the angiotensin II like immunoreactive neurons were observed within the area postrema, especially in the upper region. At the electron microscopic level, the angiotensin II-like immunoreactive cell bodies were observed as having a round, unindented nucleus. The nuclei of these neurons were not immunostained. The angiotensin II-like immunoreactive axon terminals often contained a few dense core vesicles in addition to many small clear synaptic vesicles. Numerous axon terminals were found to make synapses on immunonegative dendrites; they were also found to make synapses on angiotensin II-like immunoreactive dendrites. Many angiotensin II like immunoreactive dendrites received synapses from immunonegative axon terminals. Although angiotensin II-like immunoreactive cell bodies were sometimes postsynaptic to immunoreactive axon terminals, they did not receive synapses from immunonegative axon terminals. These results provide solid morphological evidence of AP endogenous angiotensin II and confirm that in spite of circulating angiotensin II, the local neurons in the AP may also play an important role in angiotensin II-induced cardiovascular regulation. PMID- 11020226 TI - Overflow of dopamine and cholecystokinin in rat nucleus accumbens in response to acute drug administration. AB - Cholecystokinin octapeptide (CCK-8) coexists with dopamine (DA) in rat mesencephalic neurons that project to the nucleus accumbens. To obtain indices of corelease, microdialysis probes were placed in the posterior nucleus accumbens of anesthetized rats, which were then injected acutely (s.c.) with drugs that exert known effects on DA neuronal function. Microdialysis samples were assayed for DA and CCK-8 to determine the differential overflow of these cotransmitters in response to drug treatment. Haloperidol (0.5 mg/kg), d-amphetamine (1 mg/kg), and TCP (5 mg/kg) preferentially increased DA overflow, whereas morphine (5 mg/kg) elicited marked increases in the overflow of both DA and CCK-8. These results suggest that the release of accumbal DA and CCK-8 can be differentially regulated by drug treatment. PMID- 11020227 TI - Differential modulation of frontal cortex acetylcholine by injection of substance P into the nucleus basalis magnocellularis region in the freely-moving vs. the anesthetized preparation. AB - In vivo microdialysis was used to assess the effects of unilateral substance P (SP) injection into the nucleus basalis magnocellularis on extracellular levels of acetylcholine (ACh) in the frontal cortex, either in freely moving or urethane anesthetized rats. The results show that the neurochemical effects of SP are critically dependent on the choice of the experimental preparation: In the freely moving rat, the injection procedure led to behavioral and concurrent bilateral cholinergic activation in the frontal cortex. This cholinergic activation was ipsilaterally reduced by intrabasalis injection of SP (1 ng), indicating that the peptide exerted an inhibitory influence on the neurochemical effect exerted by handling, intracranial needle insertion, and vehicle injection. In the anesthetized preparation, SP had a biphasic dose-dependent action on cortical ACh: a short-lasting ipsilateral increase immediately after injection (especially with 1 ng), and a delayed bilateral increase after more than 2 h (10, 100 ng). The procedure of inserting the injection needle moderately increased cortical ACh levels. Methodologically, these data are discussed with respect to the importance of using anesthetized vs. freely moving rats and the effects of intraparenchymal injections. PMID- 11020228 TI - Kinetics of recovery from opioids at wild-type and mutant mu opioid receptors expressed in xenopus oocytes. AB - To investigate a previous observation that classical antagonists behave as agonists at mutant H297N and H297Q mu opioid receptors, we compared the kinetics of recovery from opioids at wild-type and mutant mu receptors expressed in voltage-clamped Xenopus oocytes. The cDNA for the potassium channel GIRK1 was coinjected into the oocytes with that of the mu receptors to transduce agonist binding into a coupled electrophysiological response. The kinetics of recovery were estimated by brief test pulses of the agonist normorphine given at a frequency of 0.67 or 1 per min. After treatment with a variety of agonists, the receptors recovered from desensitization at rates that depended on the agonist, but there was little difference between mutant and wild-type receptors. Antagonists, however, induced agonist-like currents and demonstrated faster recovery at the mutant receptors. These results suggest that His-297 may comprise part of an antagonist subsite. This conclusion, when coupled with the steric theory that intrinsic activity depends on independent binary equilibration of a drug between agonist and antagonist subsites, could unify the paired observations that antagonists become agonists and recover faster at the mutant than at the wild-type receptors. Synapse 38:254-260, 2000. Published 2000 Wiley-Liss, Inc. PMID- 11020229 TI - Assessment of tyrosine hydroxylase immunoreactive innervation in five subregions of the nucleus accumbens shell in rats treated with repeated cocaine. AB - To explore the effects of behavioral sensitization on the anatomy of the nucleus accumbens shell, we employed a typical cocaine dosing paradigm and assessed tyrosine hydroxylase immunoreactive varicosities in five different areas of the shell, as well as the core of the nucleus accumbens. Rats were given bidaily injections of either saline (1 ml/kg i.p.) or cocaine (15 mg/kg i.p.) for 5 consecutive days, and sacrificed either 2 or 14 days from the last injection. Sections of the nucleus accumbens were processed for tyrosine hydroxylase immunoreactivity and the number of immunoreactive varicosities in contact with neuronal cell bodies was quantified in each of the subregions of the shell, as well as the core of the nucleus accumbens. Compared to saline controls, the cocaine-treated animals showed a significant augmentation in tyrosine hydroxylase immunoreactivity in two of the five subregions after 2 days of withdrawal in the shell, but not in the core. No differences were found in any region tested after 14 days of withdrawal. These data are the first to suggest that increases in nucleus accumbens presynaptic tyrosine hydroxylase may play a role in the development of behavioral sensitization, but not in the long-term expression of this phenomenon. PMID- 11020230 TI - Functional state of corticostriatal synapses determines their expression of short and long-term plasticity. AB - Relationships between presynaptic function and short- and long-term plasticity were investigated at adult corticostriatal synapses. Wide variability was observed in the expression of short- and long-term synaptic plasticity. Intracellular records from 47 cells produced 17 examples of LTD (<90% of control), 10 examples of no long-term change (between 90-110% of control), and 20 examples of LTP (>110% of control). Similar variation existed in paired-pulse and posttetanic plasticities. The variability expressed in all three forms of plasticity appears to be related, based on correlations found between the paired pulse ratio (PPR) and tetanus-induced short- (3 min posttetanus) and long-term plasticities (16-20 min posttetanus). These data suggest that tetanus-induced changes in synaptic strength are related to the intrinsic, preconditioned behavior of synapses. Every cell showing paired-pulse depression also expressed LTD in response to high-frequency activation of its afferents. Those synapses showing paired-pulse potentiation tended to express LTP, although exceptions did exist. Similar relationships were found in a parallel analysis of population spikes. PPR also changed in association with the expression of posttetanic and long-term depression. Greater paired-pulse potentiation was observed in medial intracellular recordings, but no medial to lateral differences were seen in posttetanic plasticities. Field recordings also showed a medial bias toward paired-pulse and posttetanic potentiation, but not in long-term plasticity. Block of postsynaptic L-type Ca(2+) channels with nifedipine eliminated LTD expression, but overall no differences were found between nifedipine and control cells. PMID- 11020231 TI - Collateral projections from the supramammillary nucleus to the medial septum and hippocampus. AB - Previous reports have shown that the supramammillary nucleus projects to the medial septum and to the hippocampus, and specifically to the dentate gyrus and the CA2/CA3a region of the hippocampus. The aim of the present study was to examine collateral projections from the supramammillary nucleus to the septum and hippocampus. The fluorescent retrograde tracers, Fluororuby and Fluorogold, were injected into regions of the septum and hippocampus, respectively, and the supramammillary nucleus was examined for the presence of single- and double labeled neurons. The main findings were: 1) pronounced numbers of single-labeled cells (about 40-60/section) were present in the supramammillary nucleus following retrograde tracer injections in either the septum or hippocampus; 2) single and double retrogradely labeled neurons were intermingled within the supramammillary nucleus and mainly localized to the lateral two-thirds of the supramammillary nucleus; 3) approximately 5-10% of supramammillary cells were double-labeled, ipsilaterally, and 2-4%, contralaterally, with injections in medial or lateral parts of the medial septum and the dentate gyrus of the hippocampus; and 4) approximately 3-5% of supramammillary cells were double-labeled, ipsilaterally, and 1-2%, contralaterally, with injections in the medial septum and CA2/CA3a of the dorsal hippocampus. Cells of the supramammillary nucleus have been shown to fire rhythmically in bursts synchronous with the hippocampal theta rhythm and have been implicated in the generation of the theta rhythm. The supramammillary cells that we identified with collateral projections to the septum and hippocampus may be directly involved in generation of the theta rhythm. PMID- 11020232 TI - Ionotropic glutamate receptor modulation preferentially affects NMDA receptor expression in rat hippocampus. AB - Electrophysiological data suggest that alterations in the function of one glutamate receptor subtype may affect the function of other subtypes. Further, previous studies have demonstrated that NMDA receptor antagonists affect NMDA and kainate receptor expression in rat hippocampus. In order to address the mutual regulation of NMDA, AMPA, and kainate receptor expression in rat hippocampus, we conducted two experiments examining the effects of NMDA and non-NMDA glutamate receptor modulators on NMDA, AMPA, and kainate receptor expression using in situ hybridization and receptor autoradiography. NMDA receptor expression was preferentially affected by systemic treatments, as all drugs significantly altered [(3)H]MK-801 binding, and several drugs increased [(3)H]ifenprodil binding. GYKI52466 and aniracetam treatments resulted in changes in both [(3)H]ifenprodil binding and NR2B mRNA levels, consistent with the association of this subunit and binding site in vitro. There were more modest effects on AMPA and kainate receptor expression, even by direct antagonists. Together, these data suggest that ionotropic glutamate receptors interact at the level of expression. These data also suggest that drug regimens targeting one ionotropic glutamate receptor subtype may indirectly affect other subtypes, potentially producing unwanted side effects. PMID- 11020233 TI - Novel in vivo electrophysiological assay for the effects of cocaine and putative "cocaine antagonists" on dopamine transporter activity of substantia nigra and ventral tegmental area dopamine neurons. AB - The aim of these studies was to establish a rapid in vivo assay for evaluating potential "cocaine antagonists," i.e., drugs postulated to block cocaine binding to the dopamine transporter (DAT) without corresponding blockade of dopamine reuptake. The assay is based on the ability of dopamine, and drugs that elevate synaptic dopamine levels, to inhibit the extracellular single unit activities of midbrain dopamine neurons in chloral hydrate-anesthetized rats. As expected, cocaine itself (0.06-16 mg/kg, i.v.) caused a dose-dependent inhibition of firing of both substantia nigra and ventral tegmental area (VTA) dopamine neurons, but had a significantly higher potency on VTA than nigral dopamine cells (ED(50)'s 1.2 and 8.8 mg/kg, respectively). VTA cells were also inhibited to a greater extent (to 4.7 +/- 4.5% vs. 41.3 +/- 6.3% of baseline rates at 16 mg/kg, respectively). We next evaluated GBR12909, a piperazine analog promoted as a "cocaine antagonist" because of its ability to bind with high affinity to the DAT, while only modestly elevating extracellular dopamine levels. The agonist- and antagonist-like properties of GBR12909 were evaluated on only VTA dopamine cells since these neurons were more fully inhibited by cocaine and have been implicated in its rewarding effects. Given alone, GBR12909 exhibited modest "cocaine-like" activity insofar as it partially inhibited VTA dopamine neurons (to 59.0 +/- 4.6% of baseline at 8 mg/kg). However, consistent with an antagonist profile, pretreatment with a low (0.5 mg/kg) dose of GBR12909, which depressed firing only slightly, resulted in a >2-fold rightward shift in the dose-response curve to cocaine (ED(50) 2.6 mg/kg). We conclude that electrophysiological testing of putative "anti-cocaine" drugs for their abilities to inhibit the firing of VTA dopamine neurons, and to block their inhibitory responses to cocaine, may provide a rapid in vivo screen for compounds expected to behave as functional cocaine antagonists in the dopamine reward system. PMID- 11020234 TI - Nicotine effects on regional cerebral blood flow in awake, resting tobacco smokers. AB - The hypothesis for this research was that regional cerebral blood flow (rCBF) would increase following nasal nicotine administration to overnight abstinent tobacco smokers in relationship to the known brain distribution of nicotinic cholinergic receptors (nAChRs). Nine male and nine female healthy adult smokers were studied. They abstained overnight from tobacco products for 10 or more hours prior to study the next morning. Nicotine nasal spray was given in doses of 1-2.5 mg total with half in each nostril while the subject was awake and resting in a supine position. Oleoresin of pepper solution in a similar volume was used as an active placebo to control for the irritating effects of nicotine. Both substances were given single blind to the subjects. Positron emission tomography (PET) with H(2)(15)O was used to measure rCBF. The data from each subject volunteer were normalized to global activity to better assess regional brain changes. Both nasal nicotine and pepper spray produced similar increases in CBF in somesthetic area II, consistent with the irritant effects of both substances. The mean rCBF effects of nasal pepper were subtracted from those of nasal nicotine to determine the actions of nicotine alone. The latter produced increases in rCBF in the thalamus, pons, Brodman area 17 of the visual cortex, and cerebellum. Some brain areas that contain a large number of nAChRs, such as the thalamus, showed an increase in CBF. Other areas that have few nAChRs, such as the cerebellum, also showed an increase in relative CBF. The hippocampal/parahippocampal areas showed greater regional decreases (left) and lesser increases (right) in CBF that correlated with the increase in plasma arterial nicotine concentrations. The results obtained indicate complex primary and secondary effects of nicotine in which only some regional brain CBF changes correlate with the known distribution of nAChR. No gender differences were noted. PMID- 11020235 TI - Role of cAMP-dependent protein kinase (PKA) in opioid agonist-induced mu-opioid receptor downregulation and tolerance in mice. AB - Studies suggest that acute and chronic opioids can regulate the cAMP-dependent protein kinase (PKA) signaling pathway and that changes in this pathway may be involved in opioid tolerance. In the present study, we examined the role of cAMP PKA on mu-opioid receptor downregulation and tolerance in mice. Mice were injected intracerebroventricular (i.c.v.) and intrathecal (i.t.) once a day with an antisense oligodeoxynucleotide directed at the mRNA for the alpha catalytic subunit of mouse PKA. Controls were treated with saline or a mismatch oligodeoxynucleotide. On day 2 of treatment, mice were implanted s.c. with a 25 mg morphine pellet and an osmotic minipump infusing morphine (40 mg/kg/day) for 3 days. Other mice were implanted with an osmotic minipump infusing etorphine (125, 250 microg/kg/day) for 2 days. Control mice were implanted s.c. with inert placebo pellets. At the end of treatment, pumps and pellets were removed and mice tested for morphine or etorphine analgesia. Other mice were sacrificed and mu opioid receptor binding assays conducted in whole brain. Both infusion doses of etorphine produced significant tolerance (ED(50) shift = 3.6 and 6.3-fold). The higher etorphine infusion produced downregulation of mu-receptor density ( approximately 30%) while the lower infusion dose of etorphine did not. Morphine treatment also produced significant tolerance in mice (ED(50) shift = 4.5-fold), but no receptor downregulation. Antisense to PKA partially blocked tolerance induced by the higher dose of etorphine, but had no effect on receptor downregulation. On the other hand, antisense to PKA completely blocked tolerance induced by morphine and the lower infusion dose of etorphine. The mismatch oligodeoxynucleotide had no effect on any measure. These results suggest that PKA has a limited role in opioid agonist-induced receptor downregulation. However, the partial block of tolerance for the high infusion dose of etorphine and the complete block of tolerance for morphine and the low infusion dose of etorphine suggests that PKA may play a critical role in tolerance that is "receptor regulation-independent." PMID- 11020236 TI - 5-HT(1A) receptor-mediated inhibition and 5-HT(2) as well as 5-HT(3) receptor mediated excitation in different subdivisions of the rat amygdala. AB - The techniques of extracellular single cell recording and microiontophoresis were used to study the effects of serotonin (5-HT) and of 5-HT(1A), 5-HT(2A/2C) and 5 HT(3) receptor agonists on the spontaneous activity of amygdaloid neurons in rats anesthetized with urethane. The background discharge rate was modified by 5-HT as well as by 5-HT agonists in about two-thirds of neurons tested in different nuclei of the amygdaloid complex. Whereas the 5-HT(2) and 5-HT(3) agonists significantly increased the neuronal discharge rate in nearly all subdivisions of the amygdala, the 5-HT(1A) agonist significantly inhibited the firing rate. Co administration of bicuculline and 5-HT receptor agonists prevented the 8-OH-DPAT induced increases in the firing rate in most cases tested, as well as the inhibitory effects of DOI or 2-methyl-5HT. Therefore, GABAergic interneurons seem to be involved in the mediation of serotonergic effects. The action of 5-HT agonists on the neuronal discharge rate was blocked by different receptor specific antagonists. The results support the hypothesis that 5-HT exerts control throughout the amygdala by acting at least on 5-HT(1A), 5-HT(2A/2C) and 5-HT(3) receptors seemingly located both on projection and interneurons. PMID- 11020237 TI - SPECT tracer [(123)I]IBZM has similar affinity to dopamine D2 and D3 receptors. AB - Emission tomography investigations of the pathophysiological involvement of the cerebral dopaminergic transmitter system in the living human brain relies heavily on a careful selection of the most suitable radioligand. In recent years, many clinical studies have employed [(123)I]IBZM in SPECT studies. The aim of the present study was to characterize the binding of IBZM to dopaminergic receptor subtypes as a means of elucidating which receptor subtypes are visualized and examined by [(123)I]IBZM. The affinity of IBZM for each of the major human dopamine receptors (D1, D2(short), D3, D4(4. 2), and D5 receptor) was determined by competitive radioligand binding assay using membranes prepared from clonal cell lines expressing the different subtypes. Radioligands with high affinity for the D1(A) and D5 receptors ([(3)H]SCH-23390), dopamine D2(short) and D4(4.2) receptors ([(3)H]Spiroperidol), and dopamine D3 receptor ([(3)H]7-OH-DPAT) were used to measure specific binding. Corresponding unlabeled displacing ligands for determination of nonspecific binding were employed. Assays were performed at 25 degrees C. These experiments show that for IBZM K(i) values were 1.6 nM for dopamine D2(s) receptors and 2.2 nM for dopamine D3 receptors. There was no binding of IBZM to D1(A), D5, or D4(4.2) receptors. In conclusion, when [(123)I]IBZM is used as SPECT tracer, the studies reflect dopaminergic D2 as well as D3 receptor binding. PMID- 11020238 TI - Distribution of the histamine H(2) receptor in monkey brain and its mRNA localization in monkey and human brain. AB - The distribution of histamine H(2) receptor mRNA was determined by in situ hybridization histochemistry in human and monkey brain. In the case of monkey brain, we combined this technique with receptor ligand autoradiography to compare the distribution of mRNA and receptor binding sites. [(125)I]Iodoaminopotentidine ([(125)I]-APT), a reversible, high specific activity antagonist with high affinity and selectivity for the H(2) receptor, was used for receptor autoradiography. Radiolabeled oligonucleotides derived from the human mRNA sequence encoding this receptor were used as hybridization probes. The highest density of the H(2) receptor mRNA in human and monkey brain was found in caudate and putamen nuclei and external layers of cerebral cortex. Moderate levels were seen in the hippocampal formation and lower densities in the dentate nucleus of cerebellum. Areas such as globus pallidus, amygdaloid complex, cerebellar cortex, and substantia nigra were devoid of hybridization signal. The distribution of H(2) receptor mRNA in monkey brain is generally in good agreement with that of the corresponding binding sites: prominent in caudate, putamen, accumbens nuclei, and cortical areas. The hippocampus showed lower densities of receptors and low levels were detected in the globus pallidus pars lateralis. No binding sites were seen in amygdaloid complex and substantia nigra. The distribution of histaminergic innervation is in good correlation with the areas of high density for H(2) receptors: caudate, putamen, and external layers of cerebral cortex in monkey and human brain. The presence of mRNA in caudate and putamen nuclei, together with its absence from substantia nigra, suggests that the H(2) receptors found in the striatum are synthesized by intrinsic cells and not by nigral dopaminergic cells. These striatal H(2) receptors may be located on short circuit striatal interneurons or somatodendritically on striatal projection neurons which project to the globus pallidus pars lateralis. In conclusion, the present results, which constitute, to our knowledge, the first report of the regional distribution of mRNA encoding H(2) receptors detected by in situ hybridization, define the sites of synthesis of H(2) receptors and are the basis for future, more detailed studies that should result in a better understanding of H(2) receptor function. PMID- 11020239 TI - Differential effects of acute and short-term lithium administration on dialysate glutamate and GABA levels in the frontal cortex of the conscious rat. AB - In the present study, we employed in vivo microdialysis in the frontal cortex of the awake rat to investigate the effects of acute and short-term (twice daily, 3 days) lithium chloride administration (1, 2, and 4 meq/kg, s.c.) on local dialysate glutamate and GABA levels. Acute lithium (1 meq/kg) failed to influence cortical glutamate levels while the higher (2 and 4 meq/kg) doses increased (+38 +/- 6% of basal levels) and reduced (-27 +/- 4%) cortical glutamate levels, respectively. Cortical GABA levels were affected by acute lithium only at the highest 4 meq/kg dose (+62 +/- 6%). Furthermore, these effects were prevented by tetrodotoxin (1 microM) and low-calcium (0.2 mM) medium perfusion. Following short-term administration, lithium increased (+58 +/- 4%) cortical dialysate glutamate levels at the 1 meq/kg dose, was ineffective at 2 meq/kg, while the effect of the 4 meq/kg dose was similar to that observed after acute administration. Interestingly, intracortical perfusion with the GABA(B) receptor antagonist CGP 35348 (100 microM) reversed the acute lithium (4 meq/kg)-induced decrease in glutamate levels. Taken together, these findings indicate a differential dose and duration dependent effect of lithium on cortical dialysate glutamate levels involving both a direct enhancement and an indirect inhibition that is mediated via an activation of local GABA(B) receptor. These findings may be relevant for the therapeutic effects of the drug. PMID- 11020240 TI - Spontaneous long-term compensatory dopaminergic sprouting in MPTP-treated mice. AB - The present study sought to determine whether severe 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) intoxication elicits spontaneous long-term compensatory sprouting in mice. Animals, once treated, were kept without further treatment for 0.5, 1, 5, or 7 months. The stability of the nigral degeneration was checked by evaluation of the number of tyrosine hydroxylase immunoreactive (TH-IR) neurons, whereas sprouting was assessed using both [(3)H]-dopamine (DA) uptake by striatal synaptosomes and optical density of TH-immunolabeled fibers in the striatum as markers. At 0.5 month after MPTP intoxication (80 mg/kg, i.p.), we observed comparable decreases of 83% in DA uptake, 83.3% in TH fiber density, and 74% in the number of TH-IR neurons compared to age-matched saline-treated animals. From 5 months onwards, both DA uptake and striatal TH fiber density increased significantly (50% and 34.9% at 5 months, 65% and 67.4% at 7 months, respectively) in comparison with age-matched saline-treated animals, although the number of TH-IR neurons remained stable (73% of degeneration at 7 months). These results indicate clearly that spontaneous long-term compensatory dopaminergic sprouting is a phenomenon that is not restricted to situations of partial nigral degeneration but can, on the contrary, constitute a response even to severe stable MPTP-induced nigral degeneration. PMID- 11020242 TI - Collagenase induction promotes mouse tumorigenesis by two independent pathways. AB - Although progress has been made in the understanding of the role of metalloproteinases in tumor progression during metastasis, little is known about their contributions, if any, to tumor formation. Accumulating evidence identified an increased presence of several matrix metalloproteinases in human cancers, but the precise role for interstitial collagenase in tumor formation or progression has not been well defined. Transient induction of collagenase was observed in wild-type mouse skin after treatment with the tumor-promoting agents 12-O tetradecanoylphorbol-13-acetate (TPA) and chrysarobin, which promote tumorigenesis through protein kinase C-dependent and -independent pathways, respectively. Transgenic mice that constitutively express interstitial collagenase within the epidermis of the skin have an increased susceptibility to tumorigenesis and produced tumors at lower doses of TPA as compared with wild type mice. Similarly, the transgenic mice showed increased tumorigenesis when promoted with chrysarobin. These results demonstrate that collagenase overexpression can contribute to tumorigenesis via protein kinase C-dependent and -independent pathways. Significantly, compared with wild-type mice, the transgenic mice demonstrated an elevated expression of c-fos in the skin at baseline, before tumor promotion, suggesting a molecular mechanism for the increased tumor susceptibility in collagenase transgenic mice. These findings further support the importance of MMP deregulation in tumorigenesis and suggest that the role of MMP family members is not limited to metastasis but may also contribute to initial tumor development. PMID- 11020241 TI - Involvement of cyclin-dependent kinases in doxorubicin-induced apoptosis in human tumor cells. AB - Apoptotic cell death caused by doxorubicin, a chemotherapeutic agent, is suppressed by expression of p21 (waf1/cip1/sdi1), a cyclin-dependent kinase (cdk) inhibitor. To examine cdk activity required for doxorubicin-induced apoptosis, we transfected p21-deficient human tumor DLD1(p21-/-) cells with plasmids carrying wild-type p21 and mutated p21 unable to bind to cdks or proliferating cell nuclear antigen. The apoptosis induced at the G(2)/M phase after doxorubicin treatment was suppressed by transient expression of the p21 with cdk-binding activity but not by the p21 lacking the activity. We also transfected cells with plasmids carrying wild-type, dominant negative and constitutively active mutants of cdk2 or cdk4. The apoptosis was suppressed by transient expression of dominant negative mutants of cdk2 or cdk4. These findings indicate that cdk is involved in the doxorubicin-induced apoptosis pathway. PMID- 11020243 TI - Reduced global genomic repair of ultraviolet light-induced cyclobutane pyrimidine dimers in simian virus 40-transformed human cells. AB - The p53 tumor-suppressor gene has been implicated in the inducible activation of excision repair of ultraviolet (UV)-induced cyclobutane pyrimidine dimers (CPDs) in human cells. Because the large T antigen (LTAg) of the simian virus 40 (SV40) binds p53 protein and can interfere with its function, it was of interest to study DNA repair in normal human fibroblasts that had been transformed by SV40 compared with that in their nontransformed parental counterparts and to determine whether such transformation attenuated global genomic repair (GGR) of CPDs. Three methods were used to measure GGR in UV-irradiated cells: (i) an immunoassay using monoclonal antibodies specific for CPDs or 6-4 photoproducts (6-4PPs), (ii) zone sedimentation in alkaline sucrose gradients to measure the average DNA strand size after specific nicking at CPD sites in duplex DNA with T4 endonuclease V (TEV), and (iii) Southern hybridization of TEV-treated DNA with strand-specific mRNA probes to assess removal of CPDs from either strand of a defined genetic sequence in an expressed gene. Whereas repair of 6-4PPs was very similar in paired SV40-transformed and primary fibroblasts, GGR of CPDs was significantly reduced in the SV40-transformed cells. In contrast, SV40 transformation did not appreciably affect the efficiency of transcription-coupled repair. These data support the hypothesis that SV40 transformation can result in reduced levels of GGR, most likely because of the inhibition of normal p53 function by LTAg. PMID- 11020244 TI - IkappaBbeta-related proteins in normal and transformed colonic epithelial cells. AB - The transcription factor nuclear factor-kappaB (NF-kappaB) regulates genes that can influence cell proliferation, apoptosis, and inflammatory responses. Since these events can contribute to carcinogenesis, we examined the expression of NF kappaB inhibitory proteins (IkappaBs) in normal and transformed colonic epithelial cells. Immunohistochemical analysis of the mouse colon revealed a high level of IkappaBbeta expression in epithelial cells relative to the rest of the tissue, whereas IkappaBalpha was found primarily in cells of the lamina propria. Mouse colon tumors showed a similar cell-specific staining pattern. Immunoblot analysis of IkappaBbeta from mouse colonocytes and the human HT-29 colon cancer cell line indicated that most of the IkappaBbeta in these cells was similar to the C-terminal-truncated IkappaBbeta2 isoform. Cell fractionation studies were consistent with IkappaBbeta being a major regulator of p65-p50 NF-kappaB complexes in HT-29 cells. Interestingly, two larger proteins specifically recognized by IkappaBbeta antibodies (p106 and p112) were found in HT-29 cells and in colon tissue of carcinogen-exposed mice. The p106 and p112 proteins bound to NF-kappaB, and their levels changed during the transient interleukin-1beta activation of NF-kappaB in HT-29 cells. Evidence was obtained indicating that p106 and p112 are stably ubiquitinated forms of IkappaBbeta. We propose that deficiencies in the proteasomal degradation of IkappaBbeta lead to p106 and p112 accumulation, which in turn alter NF-kappaB regulation in colon cancer cells. PMID- 11020245 TI - Complementation of an hMSH2 defect in human colorectal carcinoma cells by human chromosome 2 transfer. AB - The human colorectal tumor cell line LoVo has a homozygous deletion in the hMSH2 gene from exon 3 to exon 8, is deficient in mismatch repair (MMR) activity, and exhibits microsatellite instability. To determine whether the introduction of a wild type hMSH2 into LoVo restores MMR activity and stabilizes microsatellite loci, we transferred a chromosome 2 fragment containing hMSH2 into a homologous recombination-proficient chicken DT40/human hybrid (DT40 2C) and a human chromosome 2 in a mouse A9 hybrid to LoVo. Transfers of these chromosomes into LoVo resulted in LoVo both with and without a wild-type hMSH2. Complete correlation was found between the presence of the wild-type hMSH2 and hMSH2 expression, an increased stability in microsatellite loci, and competency in MMR. The hMSH2-positive LoVo hybrids also showed an increased sensitivity to N-methyl N'-nitro-N-nitrosoguanidine. This enhanced toxicity is associated with G(2) cell cycle arrest followed by premature mitosis and cell death. These results suggest that hMSH2 may be responsible for complementing mutator and drug-resistant phenotypes in chromosome 2-transferred LoVo cells. To test whether the hMSH2 in DT40 2C cells can be modified by homologous recombination, we transfected DT40 2C with a targeting vector containing an hMSH2 exon 4 disrupted by the zeocin resistant gene. The results showed that the hMSH2 locus in DT40 2C was efficiently targeted by an exogeneously transfected homologous sequence, suggesting that transfer of a modified hMSH2 from DT40 2C to LoVo via chromosome transfer could be used to determine the function of hMSH2. PMID- 11020246 TI - The language of infectious disease: a light-hearted review. PMID- 11020247 TI - Development of daptomycin for gram-positive infections. PMID- 11020248 TI - Multiplex PCR assays for the detection of clinically relevant antibiotic resistance genes in staphylococci isolated from patients infected after cardiac surgery. The ESPRIT Trial. AB - Multiresistant staphylococci (82 Staphylococcus aureus and 114 coagulase-negative staphylococci) were characterized by testing with rapid multiplex polymerase chain reaction (PCR) assays for species identification and detection of associated antibiotic resistance genes. These 196 staphylococci were isolated from 149 adult patients who developed wound infection after elective coronary artery bypass grafts and/or valve surgery. The multiplex PCR assays allowed identification of the most common staphylococcal species with S. aureus- and Staphylococcus epidermidis-specific primers as well as the detection of the erythromycin resistance genes ermA, ermB, ermC and msrA, the aminoglycoside resistance gene aac(6')-aph(2"), the oxacillin resistance gene mecA and the penicillin resistance gene blaZ. There was a very good correlation between the genotypic analysis by PCR and the phenotype determined by standard methods of susceptibility testing and identification of staphylococcal species: 100% for erythromycin resistance, 98.0% for gentamicin resistance, 99.0% for oxacillin resistance, 100% for penicillin resistance and 100% for S. aureus and S. epidermidis species identification. This study suggests that the incidence and distribution of the tested clinically relevant antibiotic resistance genes in staphylococci associated with infections after cardiac surgery do not differ from those in strains from other infections. These multiplex PCR assays may be used as diagnostic tools to replace or complement standard methods of susceptibility testing and identification of staphylococci. PMID- 11020249 TI - Evaluation of the Oxoid Aura image system for measuring zones of inhibition with the disc diffusion technique. AB - In this study the Oxoid Aura image antibiotic sensitivity test system, used as a stand-alone device, was compared with manual zone measurement and use of a template, for the determination of sensitivities. An overall correlation coefficient of 0.99 was observed for zone diameters measured using the Aura image system and zones measured manually, when the differences between zones were within 3 mm; 5.4% of zones showed a difference in zone diameter between manual and automated measurement of >3 mm. The results obtained using the template method for interpretation were less reliable than zone measurement, with cefuroxime and ampicillin tested against Enterobacteriaceae and Acinetobacter spp. When linked to a laboratory patient database, the bar code and disc identification facilities avoided errors that were associated with manual data entry. PMID- 11020250 TI - Cytotoxicity and probable mechanism of action of sulphimidazole. AB - Sulphimidazole (1-methyl-2((4-aminophenyl)-sulphonyl)-amino-5-nitroimidazole) is a new compound in which a p-aminobenzenesulphonamide radical has been attached at position 2 of the 5-nitroimidazole ring. It possesses a useful spectrum of activity in vitro against various anaerobic microorganisms and its action against aerobic and facultative bacteria is synergically enhanced in association with trimethoprim. In the present study, we determined the cytotoxicity in vitro of sulphimidazole and trimethoprim, both alone and in combination, and analysed the viability of Vero cells and the protein content of their cell lysate in the presence of increasing concentrations of these drugs. Also, in order to verify the hypothesis that the action of sulphimidazole against aerobic and facultative bacteria is mediated by the sulphonamide component of the molecule, while that against anaerobic bacteria depends on the action of the nitro group of the 5 nitroimidazole ring, we studied the mechanism of action of the new compound both indirectly, by means of microbiological techniques, and directly, by determining its oxidoreduction potential with respect to that of metronidazole. The results show that sulphimidazole is only slightly toxic in vitro for Vero cells, either alone or in association with trimethoprim, and that the combination of the two functional groups in a single molecule not only maintains its structure-activity relationship intact but also broadens its antibacterial spectrum. PMID- 11020251 TI - In vitro activity of new generation fluoroquinolones against genotypically distinct and indistinguishable Clostridium difficile isolates. AB - We compared the activities of ciprofloxacin and levofloxacin with those of the newer fluoroquinolones grepafloxacin, moxifloxacin, sparfloxacin and trovafloxacin against Clostridium difficile isolates. As there is good evidence of marked clonal spread of C. difficile, we studied both genotypically distinct (n = 26) and indistinguishable (n = 28) isolates as determined by random amplified polymorphic DNA and ribosomal spacer PCR fingerprinting. The indistinguishable strains examined represent the main UK epidemic C. difficile clone. For 17 of 54 strains (31%) we were unable to read MICs following inocula preparation using Mueller-Hinton broth. Using Schaedler's broth for inocula preparation 93% of strains had readable MICs, although geometric mean MICs were uniformly higher (2.5- to 5.4-fold) compared with results using Mueller-Hinton broth. Moxifloxacin and trovafloxacin, followed by grepafloxacin, were the most active fluoroquinolones tested and were 3- to 4-fold more active than older agents such as ciprofloxacin by both MIC methods. Unexpectedly, clonal C. difficile strains had markedly reduced susceptibility compared with the distinct strains to each of the fluoroquinolones tested. Clonal strains were more than seven-fold or 12- to 29-fold less susceptible (according to geometric mean MICs) than distinct strains to both moxifloxacin and trovafloxacin, depending on the MIC method used. It remains to be seen whether the enhanced activity of new fluoroquinolones such as moxifloxacin in comparison with other fluoroquinolones against C. difficile implies that these agents are unlikely to be associated with C. difficile infection. However, clinical use of new generation fluoroquinolones in elderly hospitalized patients where C. difficile is endemic requires further study, particularly given the reduced antibiotic susceptibility to all fluoroquinolones of the readily transmissible UK C. difficile clone. PMID- 11020252 TI - Streptococcus pneumoniae in community-acquired respiratory tract infections in Spain: the impact of serotype and geographical, seasonal and clinical factors on its susceptibility to the most commonly prescribed antibiotics. The Spanish Surveillance Group for Respiratory Pathogens. AB - Resistance of Streptococcus pneumoniae to antimicrobial agents shows geographical variation and also depends on serotype, patient age and sample origin. Factors affecting antibiotic resistance in S. pneumoniae were studied from results of a multicentre susceptibility study of 12 antimicrobial agents, carried out in 14 Spanish hospitals between May 1996 and April 1997. The most frequent serotypes were serotypes 6 (14.8%), 19 (11.8%), 23 (10.5%), 3 (9.9%), 9 (8.0%), 14 (8.0%), 15 (3.9%) and 11 (3.2%); 9.6% of isolates were not typeable. The remaining 25 serotypes constituted 20.2% of strains. Penicillin resistance was more prevalent in serotypes 14 (88%), 23 (66%), 9 (62%) and 6 (57%), whereas erythromycin resistance was more prevalent in serotypes 6 (68.5%), 15 (61.4%), 14 (49.5%), 19 (42.7%) and 23 (39.3%). Serotypes 6 and 19 were the serotypes most commonly isolated from both children and adults, although with different proportions (24.0% and 19.2% of isolates from children were serotype 6 and 19, respectively, with the corresponding figures for isolates from adults being 13.6% and 11.5%, respectively). The rates of resistance of pneumococcal strains to penicillin and cefuroxime were significantly higher in strains from children than in those from adults (resistance to penicillin, 50.4% in children and 37.0% in adults; resistance to cefuroxime, 62.4% in children and 45.6% in adults). There was significantly more resistance to erythromycin in middle ear isolates (48.9%) than in blood isolates (27%). The prevalence of resistance to beta-lactams showed a seasonal pattern, with higher rates in summer and winter, proportional to the magnitude of resistance. Susceptibility to macrolides did not vary seasonally. Our results stress the relative importance that geographical, temporal (seasonality), patient (sample type, origin, age group) and bacterium-related (serotype) factors have on the variations in susceptibility observed among different pneumococcal clinical isolates. PMID- 11020253 TI - Activity of rifapentine and its metabolite 25-O-desacetylrifapentine compared with rifampicin and rifabutin against Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis and M. bovis BCG. AB - The in vitro activity of rifapentine and its metabolite, 25-O: desacetylrifapentine, as compared with that of rifampicin and rifabutin, was determined against Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis and M. bovis BCG. MICs were determined radiometrically and by the 1% proportional method using Middlebrook 7H11 agar. The bactericidal effect of the drugs was determined in parallel at selected concentrations. For drugsusceptible isolates of M. tuberculosis, the Bactec MICs of rifapentine and 25-O:-desacetylrifapentine were 0.03-0.06 mg/L and 0. 125-0.25 mg/L, respectively. Similar MICs were obtained for M. africanum (0.03-0.125 and 0.125 0.50 mg/L, respectively), and M. bovis (0.063-0.25 and 0.125-1.0 mg/L, respectively), but MICs were considerably lower for M. bovis BCG (0.008-0.063 mg/L for rifapentine and 0.016-0.125 mg/L for its metabolite). In general, MICs determined using 7H11 agar medium were usually one or two dilutions higher than those obtained using Bactec broth. When compared with rifampicin and rifabutin, the inhibitory activity of rifapentine for drug-susceptible isolates was roughly equal to that of rifabutin, and the inhibitory activity of 25-O: desacetylrifapentine was comparable to that of rifampicin; however, rifapentine was somewhat more bactericidal than rifabutin at equal concentrations. Clinical isolates of M. tuberculosis with a high degree of resistance to rifampicin (MIC >/= 32 mg/L) were also highly resistant to rifabutin, rifapentine and 25-O: desacetylrifapentine, although the MICs of rifabutin in this case were somewhat lower than the MICs of rifapentine. PMID- 11020254 TI - Comparison of the in vitro activities of rifapentine and rifampicin against Mycobacterium tuberculosis complex. AB - The in vitro activity of rifapentine for 44 clinical isolates of Mycobacterium tuberculosis complex was compared with that of rifampicin using the Bactec radiometric method and the absolute concentration method for susceptibility testing. Twenty-nine M. tuberculosis, 11 Mycobacterium bovis and four Mycobacterium africanum strains were studied. Control tests showed that rifapentine was stable for 14 days in 7H9 broth and for 3 weeks in 7H10 agar medium. The 44 M. tuberculosis complex strains were more susceptible to rifapentine than to rifampicin, irrespective of the testing method. In the radiometric system, the MIC50 and MIC90 of rifapentine for M. tuberculosis complex strains were one or two two-fold dilutions lower than those of rifampicin (0.06-0.125 mg/L versus 0.25 mg/L, respectively). By the absolute concentration method, the MIC50 and MIC90 of rifapentine for M. tuberculosis complex strains were two two-fold dilutions lower than those of rifampicin (0.125-0.25 mg/L versus 0.5-1 mg/L, respectively). The MIC90 of rifapentine for the 44 M. tuberculosis complex strains was always 0.25 mg/L, irrespective of the method used, but the radiometric method was more reliable and more reproducible than the agar 7H10 method. PMID- 11020255 TI - Inhibition of growth of Pneumocystis carinii by lactoferrins alone and in combination with pyrimethamine, clarithromycin and minocycline. AB - The in vitro activity of lactoferrins alone and in combination with clarithromycin, minocycline and pyrimethamine was investigated against three clinical isolates of Pneumocystis carinii. Susceptibility was tested by inoculating isolates on to cell monolayers and determining the parasite count after 72 h incubation at 37 degrees C. The culture medium was supplemented with serial dilutions of each agent. At 20 mg/L, bovine lactoferrin, the most active agent, suppressed the growth of cystic and trophic forms by >60%. Human lactoferrin, at the same concentration, suppressed the growth of cystic and trophic forms by >50%. Lactoferrins at 20 mg/L combined with clarithromycin 4 mg/L had high anti-P. carinii activity, with a >90% decrease in cystic and trophic form counts. Our study suggests that lactoferrins may inhibit P. carinii growth in vitro and act synergically with other clinically used compounds. These findings lend experimental support to the use of iron-chelating agents in the therapy of pneumocystis infections. PMID- 11020256 TI - Clindamycin promotes phagocytosis and intracellular killing of periodontopathogenic bacteria by crevicular granulocytes: an in vitro study. AB - Phagocytosis of periodontopathogenic bacteria by crevicular polymorphonuclear neutrophil granulocytes (PMNs) plays a key role in the aetiology of periodontitis. Antimicrobials such as clindamycin have been proven to be effective in treating progressive forms of this disease. Therefore, the purpose of this study was to determine the effect of clindamycin on the phagocytosing properties of gingival crevicular PMNs obtained from 16 patients with rapidly progressive periodontitis (RPP), eight with localized juvenile periodontitis (LJP), 12 with adult periodontitis (AP) and 13 periodontally healthy controls. The phagocytosis assay was performed with the two strains Porphyromonas gingivalis ATCC 33277 and Actinobacillus actinomycetemcomitans Tanner FDC 44 on a slide. Phagocytosis and intracellular killing were assessed by fluorescence microscopy after staining with acridine orange. The addition of clindamycin elevated the percentage of phagocytosing PMNs in periodontitis patients and controls regardless of whether P. gingivalis or A. actinomycetemcomitans was used as test strain. In granulocytes of healthy controls an enhancement of the intracellular killing of both strains was observed if clindamycin was added. Besides the antimicrobial effect, the enhancement of the phagocytosis might be an additional indication for treatment of periodontitis patients with clindamycin. PMID- 11020257 TI - Evaluation of a human monocytic cell line THP-1 model for assay of the intracellular activities of antimicrobial agents against Legionella pneumophila. AB - We examined the intracellular activities of 11 antimicrobial agents against Legionella pneumophila using a human monocyte-derived cell line, THP-1. Colony counting and microscopic examination of L. pneumophila co-incubated with THP-1 cells (5 x 105 cells/well) were performed. Both extra- and intra-cellular multiplication of L. pneumophila were observed and were dependent on the inoculum of L. pneumophila in the culture; L. pneumophila did not grow in the cell culture medium alone. Light microscopic examination confirmed that extracellular L. pneumophila originated from THP-1 cells disrupted by bacterial multiplication. L. pneumophila multiplied by 3-4 logs after 24 h incubation with THP-1 cells and their number remained stable at 106-107 cfu/mL until 72 h. The results of viability studies using four antimicrobial agents-ciprofloxacin, erythromycin, minocycline and rifampicin-demonstrated that our system was suitable for the intracellular activity assay. We used a concept of 'minimum extracellular concentration inhibiting intracellular multiplication' (MIEC) to evaluate the intracellular activity of antimicrobial agents. The MIECs of three beta-lactams were markedly higher than their conventional MICs while those of macrolides, quinolones, rifampicin and minocycline were similar to their MICs. Our results suggest that evaluation of the clinical efficacy of drugs against L. pneumophila should include determination of their intracellular activity against the bacteria, which could be measured using our assay system in THP-1 cells. PMID- 11020258 TI - Lactoferrin given in food facilitates dermatophytosis cure in guinea pig models. AB - Dermatophytosis is the most common skin infection caused by dermatophytic fungi, such as Trichophyton spp. We studied the in vitro and in vivo antifungal effects of lactoferrin against Trichophyton. Human and bovine lactoferrin, and a bovine lactoferrin-derived peptide, lactoferricin B, showed in vitro antifungal activity that was dependent on the test strain and medium used. In guinea pigs infected on the back with Trichophyton mentagrophytes (i.e. those with tinea corporis), consecutive daily po administration of bovine lactoferrin did not prevent development of symptoms during the early phase of infection, but facilitated clinical improvement of skin lesions after the peak of the symptoms. The fungal burden in lesions was less in guinea pigs that had been given lactoferrin than in untreated controls 21 days after infection. In guinea pigs infected on the foot with T. mentagrophytes (i.e. those with tinea pedis), the fungal burden of the skin on the heel portion of the infected foot 35 days after infection was lower in animals fed lactoferrin than in controls. These results suggest the potential usefulness of lactoferrin as a food component for promoting dermatophytosis cure. PMID- 11020259 TI - Effect of supplements with lactic acid bacteria and oligofructose on the intestinal microflora during administration of cefpodoxime proxetil. AB - Thirty healthy volunteers in three groups participated in a study of the effect on the intestinal microflora of oral supplementation with Bifidobacterium longum, Lactobacillus acidophilus and oligofructose, an indigestible oligosaccharide, during oral administration of cefpodoxime proxetil bd for 7 days. Those in group A also received an oral supplement with c.1011 cfu of B. longum BB 536 and L. acidophilus NCFB 1748 and 15 g oligofructose daily, those in group B received a supplement with oligofructose only and those in group C received placebo, for 21 days. In all three groups there was a marked decrease in aerobic microorganisms, involving mainly a rapid and almost complete disappearance of Escherichia coli (P: < 0.05) during antimicrobial administration and, thereafter, an overgrowth of enterococci (P: < 0.05). The number of intestinal yeasts also increased significantly (P: < 0.05) in groups A and B over the same period. There was a dramatic decrease in anaerobic microorganisms on day 4 of administration, mainly caused by loss of bifidobacteria (P: < 0.05) in all groups. The number of lactobacilli also decreased but was significantly higher in group A than in group C at the end of cefpodoxime proxetil administration. Clostridium difficile was found in only one person from group A, but six persons each in groups B and C. Of the bifidobacterial strains isolated from the faecal samples in group A, one was similar to the strain of B. longum administered, but most volunteers were colonized by several different strains of B. longum during the investigation period. The administered strain of L. acidophilus was recovered from six patients in group A. PMID- 11020260 TI - Clarithromycin resistance stability in Helicobacter pylori: influence of the MIC and type of mutation in the 23S rRNA. AB - Thirty clarithromycin-resistant Helicobacter pylori strains (MIC range 8-64 mg/L) were subcultured in a drug-free medium and the MIC was determined every five passages to detect in vitro stability of resistance. Three out of the 30 (10%) lost their resistance after 10, 13 or 18 subcultures (MIC decrease from 8 to 0.008, from 16 to 0. 064 and from 32 to 0.016 mg/L). The effect of four macrolides at subinhibitory concentrations on the development of resistance was studied in H. pylori NCTC 11638 and TIGR 26695. A change in the MIC was observed only when NCTC11638 was exposed to 0.5 x MIC of erythromycin for 20 days. PMID- 11020261 TI - In vivo deletion of the methicillin resistance mec region from the chromosome of Staphylococcus aureus strains. AB - Two sets of Staphylococcus aureus isolates recovered from two patients exhibited similar susceptibility profiles except for oxacillin susceptibility (MSSA) or resistance (MRSA). SMA:I macrorestriction and inter-IS256 PCR analysis showed patterns closely related to the Belgian epidemic MRSA clone 1 in each pair of MSSA/MRSA strains. Loss of one large SMA:I DNA fragment and concurrent gain of a smaller fragment in the MSSA isolates was observed. The mecA sequence present in the MRSA was absent in the MSSA variant. Therefore, in vivo deletion of the mec region may occur in some lineages of S. aureus more frequently than previously thought. PMID- 11020262 TI - Antimicrobial resistance in Streptococcus pyogenes isolates in Berlin. AB - A total of 212 clinical Streptococcus pyogenes isolates were tested for susceptibility to various antibiotics by agar dilution. The overall frequency of erythromycin resistance was 12.7%, being higher in isolates from children (18.9%) than in those from adult patients (10.7%). Similar results were found for clarithromycin, while 2.8% of the isolates were resistant to ciprofloxacin. All strains were susceptible to penicillin and cefotaxime. Of the erythromycin resistant isolates subjected to the double-disc diffusion test for erythromycin and clindamycin, 35% expressed constitutive and 55% inducible resistance to clindamycin. Ten per cent were susceptible to clindamycin (M-phenotype). Thus, a high rate of macrolide resistance in S. pyogenes has emerged in Berlin. PMID- 11020263 TI - Simple 1,2-aminoalcohols as strain-specific antimalarial agents. AB - We report here the in vitro activity of a selection of 1, 2-aminoalcohol containing compounds against cloned strains of human Plasmodium falciparum. These compounds exhibit moderate antimalarial activity but a high degree of strain specificity, preferentially inhibiting a chloroquine-resistant strain of the organism. PMID- 11020264 TI - Efficacy of a novel tetracycline derivative, glycylcycline, against penicillin resistant Streptococcus pneumoniae in a mouse model of pneumonia. AB - The MIC90 of glycylcycline (< or =0.06 mg/L) against 55 strains of Streptococcus pneumoniae was 100-fold lower than that of minocycline or tetracycline. In a mouse model of penicillin-resistant S. pneumoniae (PRSP) pneumonia, glycylcycline (10 mg/kg) decreased bacterial counts in the lungs from 10(6) cfu to <10(2) cfu, whereas no apparent reduction of bacterial numbers was observed with minocycline or penicillin G. Pharmacokinetic studies showed that the half-life and area under the curve of glycylcycline were superior to those of minocycline and penicillin G in the lungs. These results show a preferential distribution of glycylcycline in the lungs and potent in vivo bactericidal activity in PRSP pneumonia. PMID- 11020265 TI - Sitafloxacin in the treatment of patients with infections caused by vancomycin resistant enterococci and methicillin-resistant Staphylococcus aureus. AB - Sitafloxacin is a new quinolone active against multi-resistant Gram-positive pathogens. An open study was conducted in patients with serious systemic infections with methicillin-resistant Staphylococcus aureus (MRSA) or vancomycin resistant enterococcus (VRE). Patients with MRSA were recruited if treatment with glycopeptides had failed. Of 11 patients with MRSA infection, four were cured, six failed treatment and one was indeterminate. Of nine patients with VRE infection (one patient had both pathogens), five were cured and four failed. Fifteen adverse events in 12 patients were potentially related to the study drug. Sitafloxacin was effective in VRE and some recalcitrant MRSA infections. PMID- 11020266 TI - Twenty months of screening for glycopeptide-intermediate Staphylococcus aureus. PMID- 11020267 TI - Trends in antimicrobial prescribing. PMID- 11020268 TI - Treatment protocols for community-acquired pneumonia: evidence-based must replace consensus-based. PMID- 11020269 TI - Clarithromycin appears to be linked with Clostridium difficile-associated diarrhoea in the elderly. PMID- 11020271 TI - Quantifying the impact of resistance for prescribers and drug developers-a function of natural resolution rates PMID- 11020270 TI - An assessment of the impact of antibiotic resistance in different bacterial species and of the contribution of animal sources to resistance in human infections. PMID- 11020272 TI - Surveillance of antimicrobial resistance among gram-negative isolates from intensive care units in eight hospitals in turkey PMID- 11020273 TI - Discovery of (R)-7-cyano-2,3,4, 5-tetrahydro-1-(1H-imidazol-4-ylmethyl)-3- (phenylmethyl)-4-(2-thienylsulfonyl)-1H-1,4-benzodiazepine (BMS-214662), a farnesyltransferase inhibitor with potent preclinical antitumor activity. AB - Continuing structure-activity studies were performed on the 2,3,4, 5-tetrahydro-1 (imidazol-4-ylalkyl)-1,4-benzodiazepine farnesyltransferase (FT) inhibitors. These studies demonstrated that a 3(R)-phenylmethyl group, a hydrophilic 7-cyano group, and a 4-sulfonyl group bearing a variety of substituents provide low nanomolar FT inhibitors with cellular activity at concentrations below 100 nM. Maximal in vivo activity in the mutated K-Ras bearing HCT-116 human colon tumor model was achieved with analogues carrying hydrophobic side chains such as propyl, phenyl, or thienyl attached to the N-4 sulfonyl group. Several such compounds achieved curative efficacy when given orally in this model. On the basis of its excellent preclinical antitumor activity and promising pharmacokinetics, compound 20 (BMS-214662, (R)-7-cyano-2,3,4, 5-tetrahydro-1-(1H imidazol-4-ylmethyl)-3-(phenylmethyl)-4-(2-thie nyl sulfonyl)-1H-1,4 benzodiazepine) has been advanced into human clinical trials. PMID- 11020274 TI - Synthesis and SAR of new 5-phenyl-3-ureido-1,5-benzodiazepines as cholecystokinin B receptor antagonists. AB - A series of 5-phenyl-3-ureidobenzodiazepine-2,4-diones was synthesized and evaluated as cholecystokinin-B (CCK-B) receptor antagonists. Structure-activity relationship (SAR) studies revealed the importance of the N-1 substituent for potent and selective CCK-B affinity. Addition of substituents at the urea side chain provided in some cases more potent compounds. Moreover the introduction of bulky substituents such as adamantylmethyl at N-1 and resolution of the racemic ureas resulted in our lead compound GV150013. PMID- 11020276 TI - Crystal structure of FMN-dependent nitroreductase from Escherichia coli B: a prodrug-activating enzyme. AB - The FMN-dependent flavoprotein nitroreductase from Escherichia coli B (NTR) is used in cancer chemotherapy to activate a range of prodrugs. The crystal structure of this enzyme has been determined, using molecular replacement methods and refined at 2.06 A resolution. The recombinant 24-kDa enzyme was crystallized in the tetragonal space group P4(1)2(1)2, with unit cell dimensions of a = b = 57.74 A and c = 275.51 A and two molecules in the asymmetric unit. The structure has a final R factor of 20.3% (R(free) = 26.7%), for all data between the resolution ranges of 10-2.06 A, and includes 4453 protein atoms, 230 water molecules, and 2 flavin mononucleotide (FMN) molecules. The functional unit is a homodimer, which forms the asymmetric unit in the crystal structure. The tertiary structures of these two monomers and their subunit interactions are nearly identical. The molecular replacement search model, the crystal structure of the major NAD(P)H:FMN oxidoreductase of Vibrio fisheri (FRase 1), was selected on the basis of its high sequence identity to that of NTR. The final superposition of these two enzymes revealed a very similar overall fold, with variation in the structures focused around surface loops and helices near the FMN cofactor. Helix G is implicated in substrate specificity and is better resolved in the present NTR structure than in the previously reported FRase 1 structure. The FMN binding pocket is also well-resolved, showing the presence of two channels leading into the active site. The amino acid side chains and main chain atoms interacting with the FMN are well-ordered. The structure of the substrate binding pocket has been used to examine substrate specificity and enzyme kinetics for prodrugs used in antibody-directed enzyme prodrug therapy (ADEPT) and gene-directed enzyme prodrug therapy (GDEPT). PMID- 11020275 TI - Replacement of glycine with dicarbonyl and related moieties in analogues of the C terminal pentapeptide of cholecystokinin: CCK(2) agonists displaying a novel binding mode. AB - Recent advances in the field of cholecystokinin have indicated the possible occurrence of multiple affinity states of the CCK(2) receptor. Besides, numerous pharmacological experiments performed "in vitro" and "in vivo" support the eventuality of different pharmacological profiles associated to CCK(2) ligands. Indeed, some agonists are essentially anxiogenic and uneffective in memory tests, whereas others are not anxiogenic and appear as able to reinforce memory. The reference compound for the latter profile is the CCK-8 analogue BC 264 (Boc Tyr(SO(3)H)-gNle-mGly-Trp-(NMe)Nle-Asp-Phe-NH(2)). However, although tetrapeptide ligands based on CCK-4 (Trp-Met-Asp-Phe-NH(2)) are known to possess sufficient structural features for CCK(2) recognition, none shares the properties of BC 264. Hence we have developed new short peptidic or pseudo-peptidic derivatives containing the C-terminal tetrapeptide of BC 264. Our results indicate that some compounds characterized by the presence of two carbonyl groups at the N-terminus, as in 2b (HO(2)C-CH(2)-CONH-Trp-(NMe)Nle-Asp-Phe-NH(2)), are likely to show a BC 264-like profile, bind to the CCK(2) receptor in a specific way, and display remarkable affinities (2b: 0.28 nM on guinea-pig cortex membrane preparations). This original binding mode is discussed and further enlightened by NMR and molecular modeling studies. PMID- 11020277 TI - Design, synthesis, and SAR of novel carbapenem antibiotics with high stability to Xanthomonas maltophilia oxyiminocephalosporinase type II. AB - Racemic cis-6-(phenylacetamido)carbapenem (21), 2-hydroxycarbonyl-cis-6 (phenylacetamido)carbapenem (22), 2-methoxycarbonyl-cis-6 (phenylacetamido)carbapenem (30), 2-methoxycarbomethyl-cis-6 (phenylacetamido)carbapenem (33), 2-hydroxyethyl-cis-6 (phenylacetamido)carbapenem (34), and 2-acetoxyethyl-cis-6 (phenylacetamido)carbapenem (35) were synthesized. Formation of the carbapenem nuclei in 21, 22, and 30 involved dehydrophosphonation of the corresponding 2 diphenylphosphono-6-(phenylacetamido)carbapenam precursors 14, 15, and 28 using trimethylsilyl triflate and 1,8-diazabicyclo[5.4.0]undec-7-ene in THF. Syntheses of carbapenems 33-35 involved a Wittig reaction of carbapenam 14 with methyl glyoxylate in the presence of lithium 2,2,6,6-tetramethylpiperidine in THF. For the antibacterial activities against Staphylococcus aureus FDA 209P, S. aureus 95, Escherichia coli ATCC 39188, Klebsiellapneumoniae NCTC 418, Pseudomonas aeruginosa 1101-75, and P. aeruginosa 18S-H, carbapenems (+/-)-21, (+/-)-22, (+/ )-30, and (+/-)-33-35 were found comparable with imipenem ((+)-3), yet they were notably more potent than (+)-3 against Xanthomonas maltophilia GN 12873. On the other hand, unlike (+)-3, carbapenems (+/-)-21, (+/-)-22, (+/-)-30, and (+/-)-33 35 were stable to X. maltophilia oxyiminocephalosporinase type II. Their beta lactamase inhibitory properties, however, were found to be more comparable with those of penicillin G ((+)-4) than to those of imipenem ((+)-3). A combination of imipenem ((+)-3) with (+/-)-21, (+/-)-22, (+/-)-30, and (+/-)-33-35 resulted in synergistic antibacterial activity against X. maltophilia GN 12873. Results from the biological tests were correlated with the distribution of the electron density at C(2)=C(3) of carbapenems upon reaction with transpeptidases or beta lactamases. PMID- 11020278 TI - A peptide prodrug approach for improving bisphosphonate oral absorption. AB - This work was aimed at improving the absorption of bisphosphonates by targeting carrier systems in the intestine and the intestinal peptide carrier system (hPEPT1), in particular. (14)C-Labeled pamidronate and alendronate as well as radiolabeled and "cold" peptidyl-bisphosphonates, Pro-[(3)H]Phe [(14)C]pamidronate, and Pro-[(3)H]Phe-[(14)C]alendronate were synthesized. In situ single-pass perfusion studies revealed competitive inhibition of transport by Pro-Phe, suggesting peptide carrier-mediated transport. Prodrug transport in the Caco-2 cell line was significantly better than that of the parent drugs, and the prodrugs exhibited high affinity to the intestinal tissue. Oral administration of the dipeptidyl prodrugs resulted in a 3-fold increase in drug absorption following oral administration in rats, and the bioavailability of Pro Phe-alendronate was 3.3 (F(TIBIA)) and 1.9 (F(URINE)) times higher than that of the parent drug. The results indicate that the oral absorption of bisphosphonates can be improved by peptidyl prodrugs via the hPEPT1; however, other transporters may also be involved. PMID- 11020279 TI - New substituted 1-(2,3-dihydrobenzo[1, 4]dioxin-2-ylmethyl)piperidin-4-yl derivatives with alpha(2)-adrenoceptor antagonist activity. AB - The emergence of a novel theory concerning the role of noradrenaline in the progression and the treatment of neurodegenerative diseases such as Parkinson's and Alzheimer's diseases has provided a new impetus toward the discovery of novel compounds acting at alpha(2)-adrenoceptors. A series of substituted 1-(2, 3 dihydrobenzo[1,4]dioxin-2-ylmethyl)piperidin-4-yl derivatives bearing an amide, urea, or imidazolidinone moiety was studied. Some members of this series of compounds proved to be potent alpha(2)-adrenoceptor antagonists with good selectivity versus alpha(1)-adrenergic and D(2)-dopamine receptors. Particular emphasis is given to compound 33g which displays potent alpha(2)-adrenoceptor binding affinity in vitro and central effects in vivo following oral administration. PMID- 11020280 TI - Synthesis, structure, and antitumor activity of a novel tetranuclear titanium complex. AB - The coordination complex cyclo-tetrakis[bis(1-phenyl-3-methyl-4-benzoylpyrazolon 5-ato++ +)mu-o xotitanium(IV)] has been synthesized and characterized with IR and NMR spectroscopies and X-ray diffraction. The core of this species consists of an eight-membered Ti-mu-oxo ring with alternate short-long Ti-O bond lengths. Besides these two O ligands, each metal is bound octahedrally to four O atoms from two chelating 1-phenyl-3-methyl-4-benzoylpyrazolon-5-ato anions. Several sets of Ti-O bond lengths are present: the shortest are the two Ti-O(oxo) (which are cis to each other), the longest are the two Ti-O(acyl) (cis to each other), and the two Ti-O(pyrazolonato) (trans to each other) are intermediate. The beta diketonate ligand asymmetry, a feature considered essential in other antitumor Ti compounds, induces the short-long Ti-O(oxo) sequence of bond lengths. The antitumor activity of this compound, encapsulated in a dipalmitoylphosphatidylcholine liposome, has been studied in vitro using TA-3 (mouse mammary adenocarcinoma), HEP-2 (human epithelial larynx carcinoma), and VERO (African green monkey kidney) cell lines and in vivo in CF-1 and AJ female mice ip inoculated with TA-3. In vitro cytotoxicity is greater for TA-3 than for HEP-2 and null for VERO cell lines. In vivo results show a marked increase in survival time (T/C = 293% for AJ and 208% for CF-1), whereas tumor weight decrease was observed for CF-1-treated mice. These results suggest the Ti complex liposome system may be promising as an antitumor drug. PMID- 11020281 TI - Synthesis and hypolipidemic and antiplatelet activities of alpha-asarone isomers in humans (in vitro), mice (in vivo), and rats (in vivo). AB - A series of alpha-asarone isomers was synthesized and investigated for their hypolipidemic and antiplatelet activity. Considering the hypolipidemic activity in rats at a dose of 80 mg/kg/day, some isomers were more potent than clofibrate at 150 mg/kg. Compound 3 was one of the most active agents elevating the HDL cholesterol level by 56% and lowering the LDL cholesterol level by 46.8% in rats after 7 days of administration. The activities of the platelet aggregation test in vitro were significant but lower than those of the reference substances (indomethacine and acetylsalicylic acid). In the pulmonary thromboembolic in vivo test in mice, two compounds (alpha-asarone (6) and compound 4) produced significant antithrombotic effects at 100 mg/kg, namely 44% and 52% protection against lung microembolia, respectively. alpha-Asarone derivatives form a new group of potential hypolipidemic and/or antithrombotic agents. The compounds 3, 4, and 6 may serve as lead substances whose structural modifications may result in original drugs. PMID- 11020282 TI - Carbonic anhydrase and matrix metalloproteinase inhibitors: sulfonylated amino acid hydroxamates with MMP inhibitory properties act as efficient inhibitors of CA isozymes I, II, and IV, and N-hydroxysulfonamides inhibit both these zinc enzymes. AB - The 14 different carbonic anhydrase (CA, EC 4.2.1.1) isozymes as well as the 23 different matrix metalloproteinases (MMPs) isolated up to now in higher vertebrates play important physiological functions in these organisms. Unsubstituted sulfonamides act as high-affinity inhibitors for the first type of these enzymes, whereas hydroxamates strongly inhibit the latter ones. Since the active site geometry around the zinc ion in these two types of metalloenzymes is rather similar, we tested whether sulfonylated amino acid hydroxamates of the type RSO(2)NX-AA-CONHOH (X = H, benzyl, substituted benzyl; AA = amino acid moiety, such as Gly, Ala, Val, Leu) with well-known inhibitory properties against MMPs and Clostridium histolyticum collagenase (ChC, another zinc enzyme related to the MMPs) might also act as CA inhibitors. We also investigated whether N hydroxysulfonamides of the type RSO(2)NHOH (which are effective CA inhibitors) inhibit MMPs and ChC. Here we report several potent sulfonylated amino acid hydroxamate CA inhibitors (with inhibition constants in the range of 5-40 nM, against the human isozymes hCA I and hCA II, and 10-50 nM, against the bovine isozyme bCA IV), as well as preliminary SAR for this new class of non-sulfonamide CA inhibitors. Some N-hydroxysulfonamides also showed inhibitory properties (in the micromolar range) against MMP-1, MMP-2, MMP-8, MMP-9, and ChC. Thus, the SO(2)NHOH group is a new zinc-binding function for the design of MMP inhibitors. Both CA as well as MMPs are involved, among others, in carcinogenesis and tumor invasion processes. On the basis of these findings, we suggest that the mechanism of antitumor action with some hydroxamate inhibitors might also involve inhibition of some CA isozymes (such as CA IX, CA XII, and CA XIV) present only in tumor cell membranes, in addition to collagenases/gelatinases of the MMP type. Our data also suggest that it should be possible to develop dual enzyme inhibitors that would strongly inhibit both these metalloenzymes, CAs and MMPs, based on the nature of the R, AA, and X moieties in the above formula. Compact X (such as H) and AA (such as Gly) moieties favor CA over MMP inhibition, whereas bulkier X (benzyl, substituted benzyl, etc.) and AA (such as Val, Leu) moieties and substituted-aryl R groups are advantageous for obtaining potent MMP and ChC inhibitors, which show lower affinity for CA. PMID- 11020283 TI - Synthesis of new indeno[1,2-c]isoquinolines: cytotoxic non-camptothecin topoisomerase I inhibitors. AB - In an attempt to design and synthesize potential anticancer agents acting by inhibition of topoisomerase I (top1), a new series of indenoisoquinolines was prepared and tested for cytotoxicity in human cancer cell cultures and for activity against top1. The synthesis relied on the condensation of substituted Schiff bases with homophthalic anhydrides to produce cis-3-aryl-4 carboxyisoquinolones that were cyclized to indenoisoquinolines in the presence of thionyl chloride. Both top1 inhibitory activity and cytotoxicity maximized in a single compound, 6-[3-(2-hydroxyethyl)aminopropyl]-5,6-dihydro-2,3-dimethoxy-8, 9 methylenedioxy-5,11-dioxo-11H-indeno[1,2-c]isoquinoline hydrochloride (19a), which proved to be a very potent top1 inhibitor having a 110 nM mean graph midpoint (MGM) when tested for cytotoxicity in 55 human cancer cell cultures. A number of structurally related indenoisoquinolines were also obtained that had both potent cytotoxicity as well as top1 inhibitory activity. The key feature of the more potent compounds was the presence of an aminoalkyl side chain on the indenoisoquinoline nitrogen atom. The DNA cleavage patterns induced by top1 in the presence of the indenoisoquinolines were different from those seen with camptothecin. Some of the cleavage sites induced by the indenoisoquinolines were different from those seen with camptothecin, and conversely, camptothecin induced unique cleavage sites not apparent with the indenoisoquinolines. However, both camptothecin and the indenoisoquinolines also induced DNA cleavage sites that were the same in both series but varied in intensity. In addition, some of the DNA cleavages seen with the free base of 19a (compound 18c) in the presence of top1 were inhibited at higher drug concentrations, suggesting either a direct inhibition of the enzyme or an alternative mechanism involving DNA intercalation. Consistent with intercalation, compound 18c did unwind DNA. PMID- 11020284 TI - Deriving a quantitative chirality measure from molecular similarity indices. AB - A versatile new method has been developed as a continuous symmetry measure for chiral compounds. The application of principal component analysis (PCA) to the complete N x N pairwise similarity matrices (electrostatic potential and shape indices) of a series of dihydropyridine calcium channel antagonists allowed to single out a chirality component and to compute a chirality score in terms of the between-enantiomers difference on the component value. The possibility to have chirality defined continuously at the series level could be of importance in eudismic analyses where the relative potency of two enantiomers is studied as well as in QSAR studies dealing with chiral molecules in order to improve the power of the generated models. PMID- 11020285 TI - Synthesis and cytotoxicity of 4-amino-5-oxopyrido[2,3-d]pyrimidine nucleosides. AB - A number of nucleoside analogues have been either used clinically as anticancer drugs or evaluated in clinical studies, while new nucleoside analogues continue to show promise. In this article, we report synthesis and cytotoxicity of a series of new pyrido[2, 3-d]pyrimidine nucleosides. 2-Amino-3-cyano-4 methoxypyridine was converted, in two steps, to 4-amino-5-oxopyrido[2,3 d]pyrimidine. A variety of 1-O-acetylated pentose sugar derivatives were condensed with silylated 4-amino-5-oxopyrido[2,3-d]pyrimidine, followed by protection, to afford a series of 4-amino-5-oxopyrido[2, 3-d]pyrimidine nucleosides. Further derivatizations provided an additional group of pyrido[2,3 d]pyrimidine nucleosides. These nucleosides were evaluated for in vitro cytotoxicity to human prostate cancer (HTB-81) and mouse melanoma (B16) cells as well as normal human fibroblasts (NHF). A number of compounds (1a,b, 2a-c,f, 3f+4d) showed significant cytotoxicity to cancer cells, with 4-amino-5-oxo-8 (beta-D-ribofuranosyl)pyrido[2,3-d]pyrimidine (1b) being the most potent proliferation inhibitor (EC(50): 0.06-0.08 microM) to all types of cells tested. However, a selective inhibition to the cancer cells was observed for 4-amino-5 oxo-8-(beta-D-xylofuranosyl)pyrido[2,3-d]pyrimidine (2b), which is a potent inhibitor of HTB-81 (EC(50): 0.73 microM) and has a favorable in vitro selectivity index (28). PMID- 11020286 TI - Fast calculation of molecular polar surface area as a sum of fragment-based contributions and its application to the prediction of drug transport properties. AB - Molecular polar surface area (PSA), i.e., surface belonging to polar atoms, is a descriptor that was shown to correlate well with passive molecular transport through membranes and, therefore, allows prediction of transport properties of drugs. The calculation of PSA, however, is rather time-consuming because of the necessity to generate a reasonable 3D molecular geometry and the calculation of the surface itself. A new approach for the calculation of the PSA is presented here, based on the summation of tabulated surface contributions of polar fragments. The method, termed topological PSA (TPSA), provides results which are practically identical with the 3D PSA (the correlation coefficient between 3D PSA and fragment-based TPSA for 34 810 molecules from the World Drug Index is 0.99), while the computation speed is 2-3 orders of magnitude faster. The new methodology may, therefore, be used for fast bioavailability screening of virtual libraries having millions of molecules. This article describes the new methodology and shows the results of validation studies based on sets of published absorption data, including intestinal absorption, Caco-2 monolayer penetration, and blood-brain barrier penetration. PMID- 11020287 TI - Benzo[c]quinolizin-3-ones: a novel class of potent and selective nonsteroidal inhibitors of human steroid 5alpha-reductase 1. AB - The synthesis and biological evaluation of a series of novel, selective inhibitors of isoenzyme 1 of human 5alpha-reductase (5alphaR) (EC 1.3.99.5) are reported. The inhibitors are 4aH- (19-29) or 1H-tetrahydrobenzo[c]quinolizin-3 ones (35-47) bearing at positions 1, 4, 5, and 6 a methyl group and at position 8 a hydrogen, methyl group, or chlorine atom. All these compounds were tested toward 5alphaR-1 and 5alphaR-2 expressed in CHO cells (CHO 1827 and CHO 1829, respectively) resulting in selective inhibitors of the type 1 isoenzyme, with inhibitory potencies (IC(50)) ranging from 7.6 to 9100 nM. The inhibitors of the 4aH-series, having a double bond at position 1,2, were generally less active than the corresponding inhibitors of the 1H-series having the double bond at position 4,4a on the A ring. The presence of a methyl group at position 4 (as in compounds 39-40 and 45-47), associated with a substituent at position 8, determined the highest inhibition potency (IC(50) from 7.6 to 20 nM). Compounds 39 and 40, having K(i) values of 5.8+/-1.8 and 2.7+/-0.6 nM, respectively, toward 5alphaR-1 expressed in CHO cells, were also tested toward native 5alphaR-1 in human scalp and 5alphaR-2 in human prostate homogenates, in comparison with finasteride and the known 5alphaR-1-selective inhibitor LY191704, and their mechanism of inhibition was determined. They both inhibited the enzyme through a reversible competitive mechanism and again were selective inhibitors of 5alphaR-1 with IC(50) values of 41 nM. These specific features make these inhibitors suitable candidates for further development as drugs in the treatment of DHT-dependent disorders such as acne and androgenic alopecia in men and hirsutism in women. PMID- 11020288 TI - Nonpeptide alpha(v)beta(3) antagonists. 1. Transformation of a potent, integrin selective alpha(IIb)beta(3) antagonist into a potent alpha(v)beta(3) antagonist. AB - Modification of the potent fibrinogen receptor (alpha(IIb)beta(3)) antagonist 1 generated compounds with high affinity for the vitronectin receptor alpha(v)beta(3). Sequential modification of the basic N-terminus of 1 led to the identification of the 5,6,7, 8-tetrahydro[1,8]naphthyridine moiety (THN) as a lipophilic, moderately basic N-terminus that provides molecules with excellent potency and selectivity for the integrin receptor alpha(v)beta(3). The THN containing analogue 5 is a potent inhibitor of bone resorption in vitro and in vivo. In addition, the identification of a novel, nonpeptide radioligand with high affinity to alpha(v)beta(3) is also reported. PMID- 11020290 TI - Synthesis of novel 3,7-substituted-2-(3',4'-dihydroxyphenyl)flavones with improved antioxidant activity. AB - A series of 3,7-disubstituted-2-(3',4'-dihydroxyphenyl)flavones was synthesized as potential cardioprotective agents in doxorubicin antitumor therapy. The influence of substituents on the 3 and 7 positions of the flavone nucleus on radical scavenging and antioxidant properties was explored to improve the antioxidant activity of our lead compound monoHER. In the TEAC assay most compounds had a similar potency (3.5-5 times as potent as trolox), but in the LPO assay IC(50) values ranged from 0.2 to 37 microM. In general, the 3-substituted flavones (9a-j) were the most potent compounds in the LPO assay. The number of hydroxyl groups is not the only prerequisite for antioxidant activity. Substitution in ring A of the flavonoid is not necessary for high activity, but the presence of a 7-OH group significantly modifies the antioxidant activity. The compounds are good antioxidants, which makes it interesting to evaluate them as cardioprotective agents. PMID- 11020289 TI - Synthesis and opioid receptor affinity of a series of 2, 4-diaryl-substituted 3,7 diazabicylononanones. AB - 3,7-Diazabicyclo[3.3.1]nonan-9-ones having aryl rings in positions 2 and 4 with systematically varied substituents were synthesized using a double Mannich procedure. Radioligand binding assays were performed to measure the affinity of the compounds to the mu-, delta-, and kappa-opioid receptors. The affinity of all 2, 4-diphenyl-substituted 3,7-diazabicyclo[3.3.1]nonan-9-ones to the mu- and delta-receptors was found to be low. In contrast, with exception of the nitro- and cyanophenyl-substituted compounds, most of the diazabicycles showed considerable affinity for the kappa-receptor. In particular, the m-fluoro-, p methoxy-, and m-hydroxy-substituted compounds have an affinity in the submicromolar range. Due to solubility problems in aqueous media, salts of HZ2 were synthesized. The methiodide shows high kappa-affinity and may, thus, be a promising candidate for development of a peripheral kappa-agonist, e.g. for use in the case of rheumatoid arthritis. PMID- 11020291 TI - New arylpiperazine derivatives as antagonists of the human cloned 5-HT(4) receptor isoforms. AB - New derivatives of arylpiperazine 9 were designed from ML 10302, a potent 5-HT(4) receptor agonist in the gastrointestinal system. Compounds were synthesized by condensation of a number of available arylpiperazines or heteroarylpiperazines with 2-bromoethyl 4-amino-5-chloro-2-methoxybenzoate. They were evaluated in binding assays on the recently cloned human 5-HT(4(e)) isoform stably expressed in C6 glial cells with [(3)H]GR 113808 as the radioligand. The affinity values (K(i)) depended upon the substituent on the aromatic ring. A chlorine atom produced a marked drop in activity (K(i) > 100 nM), while a m-methoxy group gave a compound with nanomolar affinity (K(i) = 3 nM). The most potent compounds were the heterocyclic derivatives with pyrimidine, pyrazine, pyridazine, or pyridine moieties (compounds 9r, 9t, 9u, 9x, respectively). K(i) values for 9a and 9r were determined for the 5-HT(4(a)), 5-HT(4(b)), 5-HT(4(c)), and 5-HT(4(d)) receptor isoforms transiently expressed in COS cells. The results indicated that the compounds were not selective. They produced an inhibition of the 5-HT-stimulated cyclic AMP synthesis in the C6 glial cells stably expressing the 5-HT(4(e)) receptor and shifted the 5-HT concentration-effect curve on adenylyl cyclase activity with pK(D) values of 7.44 and 8.47, respectively. In isolated human atrial myocytes, 9r antagonized the stimulatory effect of 5-HT on the L-type calcium current (I(Ca)) with a K(D) value of 0.7 nM. PMID- 11020292 TI - beta-Turned dipeptoids as potent and selective CCK(1) receptor antagonists. AB - To improve our knowledge of the bioactive conformation of CCK(1) antagonists, we previously described that replacement of the alpha-MeTrp residue of dipeptoids with the (2S,5S, 11bR)-2-amino-3-oxohexahydroindolizino[8,7-b]indole-5-carbox ylate (IBTM) skeleton, a probed type II' beta-turn mimetic, led to restricted analogues (2S,5S,11bR,1'S)- and (2S,5S,11bR, 1'R)-2-(benzyloxycarbonyl)amino-5 [1'-benzyl-2'-(carboxy)ethyl]carbam oyl-3-oxo-2,3,5,6,11,11b-hexahydro-1H indolizino[8,7-b]indole, 1a,b, showing high binding affinity and selectivity for CCK(1) receptors. In this report, we describe the synthesis and binding profile of new analogues of compounds 1 designed to explore the importance of the C terminal residue and of the type of beta-turn on the receptor binding affinity and selectivity. Structure-affinity relationship studies show that a C-terminal free carboxylic acid and an S configuration of the Phe and betaHph residues are favorable for CCK(1) receptor recognition. Moreover, selectivity for this receptor subtype is critically affected by the beta-turn type. Thus, while compounds 15a and 16a, containing the (2S,5S,11bR)- and (2R,5R, 11bS)-IBTM frameworks, respectively, are both endowed with nanomolar affinity for CCK(1) receptors, restricted dipeptoid derivative 15a, incorporating the type II' IBTM mimetic, shows approximately 6-fold higher CCK(1) selectivity than analogue 16a, with the type II mimetic. From these results, we propose that the presence of a beta-turn-like conformation within the peptide backbone of dipeptoids could contribute to their bioactive conformation at the CCK(1) receptor subtype. Concerning functional activity, compounds 15a and 16a behave as CCK(1) receptor antagonists. PMID- 11020293 TI - Novel cannabinol probes for CB1 and CB2 cannabinoid receptors. AB - The observation that the phenolic hydroxyl of THCs was important for binding to the CB1 receptor but not as critical for binding to the CB2 receptor prompted us to extend this finding to the cannabinol (CBN) series. To study the SAR of CBN analogues, CBN derivatives with substitution at the C-1, C-3, and C-9 positions were chosen since these positions have played a key role in the SAR of THCs. CBN 3-(1',1'-dimethylheptyl) analogues were prepared by sulfur dehydrogenation of Delta(8)-THC-3-(1',1'-dimethylheptyl) analogues. 9-Substituted CBN analogues were prepared by the standard sulfur dehydrogenation of 9-substituted Delta(8)-THC analogues (Scheme 1), which in turn were prepared following our previous procedure using selenium dioxide oxidation of the corresponding Delta(8)-THCs followed by sodium chlorite oxidation to give the 9-carboxy-Delta(8)-THC derivatives. 11-Hydroxy-CBN analogues were prepared from the corresponding 9 carbomethoxy-CBN analogues by reduction with LiAlH(4). Deoxy-CBN analogue 14 was prepared from the corresponding Delta(8)-THC analogue 11 by conversion of the phenolic hydroxyl to the phosphate derivative 12, followed by lithium ammonia reduction to provide the deoxy-Delta(8)-THC analogue 13, which in turn was dehydrogenated with sulfur to provide the deoxy-CBN analogue 14 (Scheme 2). The various analogues were assayed for binding both to the brain and the peripheral cannabinoid receptors (CB1 and CB2). We have found that the binding profile differs widely between the CBN and the THC series. Specifically, in the CBN series the removal of the phenolic hydroxyl decreases binding affinity to both the CB1 and CB2 receptors, whereas in the THC series, CB1 affinity is selectively reduced. Thus, in the CBN series, the selectivity of binding observed with the removal of the hydroxy group is decreased severalfold as compared to what occurs in the THC series. Generally, high affinity for the CB2 receptor was found in analogues when the phenolic hydroxyl was present. The 3-(1', 1'-dimethylheptyl) derivatives were found to have much higher affinities than the CBN analogues, which is in complete agreement with previously reported work by Rhee et al. PMID- 11020294 TI - Binding of a diverse set of ligands to avidin and streptavidin: an accurate quantitative prediction of their relative affinities by a combination of molecular mechanics and continuum solvent models. AB - We report calculations of free energies of binding, DeltaG(bind), between a diverse set of nine ligands and avidin as well as between a peptide and streptavidin using the recently developed MM/PBSA approach. This method makes use of a molecular dynamics simulation of the ligand-protein complex to generate a thermally averaged ensemble of conformations of the molecules that are involved in the complex formation. Based on this set of structures, a free energy of binding is calculated using molecular mechanical and continuum solvent energies as well as including estimates of the nonpolar solvation free energy and solute entropy. We compare in our simulations different classes of ligands, involving biotin derivatives, the dye 2-(4'-hydroxyazobenzene)benzoic acid (HABA), and a cyclic hexapeptide, which cover a large range of binding free energies from -5 to -20 kcal/mol. Our calculations are able to reproduce experimental DeltaG(bind) values with a very good correlation coefficient of r(2) = 0.92. This agreement is considerably better than the results obtained with an alternate approach, the linear interaction energy approximation, for this system (r(2) = 0.55). PMID- 11020295 TI - Synthesis of new 2,6-prolylxylidide analogues of tocainide as stereoselective blockers of voltage-gated Na(+) channels with increased potency and improved use dependent activity. AB - A series of tocainide chiral analogues were designed, synthesized, and evaluated in vitro, in pure enantiomeric form, as use-dependent blockers of skeletal muscle sodium channels to better understand the structural requirements responsible for the antimyotonic activity. The voltage clamp recordings showed a remarkable increase of both potency and use-dependent behavior with the analogue N-(2, 6 dimethylphenyl)-2-pyrrolidinecarboxamide (1a). In fact (R)-1a was 5-fold more potent than (R)-tocainide in producing the tonic block, i.e., the reduction of peak sodium current in resting conditions after application of the compound, but it was 21-fold more potent in condition of high frequency of stimulation (phasic block). Furthermore, as opposite to tocainide, this compound was also stereoselective, (S)-1a being 2-3-fold less potent than (R)-1a. The introduction in 1a of a methyl group in place of the hydrogen bonded to either the aminic nitrogen atom [N-(2, 6-dimethylphenyl)-1-methyl-2-pyrrolidinecarboxamide (2a)] or the amidic nitrogen atom [N-(2, 6-dimethylphenyl)-N-methyl-2 pyrrolidinecarboxamide (3a)] led unexpectedly to an inversion of stereoselectivity, the (S)-enantiomers being 3-fold more potent than the (R) ones. The comparison between eutomers showed that (S)-2a and (S)-3a are almost equieffective to (R)-1a in producing a tonic block, the half-maximal concentrations being about 100 microM; however, the use-dependent behavior was remarkably decreased by the presence of the methyl group: i.e., the gain of potency observed at high frequency of stimulation amounted to 3 and 1.6 times for 2a and 3a, respectively. The replacement of both hydrogens bonded to the aminic and amidic nitrogen atoms resulted in N-(2,6-dimethylphenyl)-N, 1-dimethyl-2 pyrrolidinecarboxamide (4a) in which the (S)-isomer was still twice as potent as the (R)-one, but the absolute potency and mostly the use-dependent behavior were strongly reduced, showing therefore no clear advantages with respect to tocainide. The use-dependent behavior, which plays a pivotal role for antimyotonic activity, is strongly reduced by the presence of methyl groups on the nitrogen atoms, likely for modification of pK(a) and/or for constraint of molecular conformation. PMID- 11020296 TI - 6-Aminoquinolones as new potential anti-HIV agents. AB - A series of 6-aminoquinolone compounds were evaluated for their in vitro activity against human immunodeficiency virus type 1 (HIV-1). Compound 12a, bearing a methyl substituent at the N-1 position and a 4-(2-pyridyl)-1-piperazine moiety at the C-7 position, was the most active in inhibiting HIV-1 replication on de novo infected C8166 human lymphoblastoid cell lines. The 12a EC(50) value was 0.1 microM, a 7-20-fold lower concentration relative to that for compounds 8a and 7a containing a cyclopropyl and tert-butyl substituent at the N-1 position, respectively. When the C-6 amino group was replaced with a fluorine atom, a decreased antiviral effect was observed. The observed effects are selective, since potency is substantially reduced when testing the compounds against the herpes simplex virus type 1 (HSV-1). Active quinolone derivatives very efficiently interact with TAR RNA, which suggests a nucleic acid-targeted mechanism of action. PMID- 11020297 TI - Melanoma peptide MART-1(27-35) analogues with enhanced binding capacity to the human class I histocompatibility molecule HLA-A2 by introduction of a beta-amino acid residue: implications for recognition by tumor-infiltrating lymphocytes. AB - The design of heteroclytic antigens with high MHC binding capacity is of particular interest to overcome the weak immunogenicity of peptide epitopes derived from tissue antigens expressed by tumors. In the present study, double substituted peptide analogues of the tumor-associated antigen MART-1(27-35) incorporating a substitution at a primary anchor residue and a beta-amino acid residue at different positions in the sequence were synthesized and evaluated for binding to the human histocompatibility class I molecule HLA-A2 and for recognition by tumor-infiltrating lymphocytes. Interestingly, by combining a Leu for Ala substitution at P2 (which alone is deleterious for antigenic activity) with a beta-amino acid substitution at a putative TCR contact residue, recognition by tumor-infiltrating lymphocytes was partially restored. The analogue [Leu(28),beta-HIle(30)]MART-1(27-35) displays both a higher affinity to HLA-A2 and a more prolonged complex stability compared to [Leu(28)]MART-1(27-35). Overall, these results suggest that double-substitution strategies and beta-amino acid replacements at putative TCR contact residues might prove useful for the design of epitope mimics with high MHC binding capacity. PMID- 11020299 TI - Molecular phylogeny and biogeography of the nasute termite genus Nasutitermes (Isoptera: Termitidae) in the pacific tropics. AB - The nasute termite genus Nasutitermes is widely distributed over all tropical regions. The phylogenetic relationships among 17 Nasutitermes species from the Pacific tropics were inferred from sequences of mitochondrial cytochrome oxidase II and 16S ribosomal RNA genes. Several methods of analysis yielded phylogenetic trees showing almost the same topology and in good agreement with reconstructions based on morphological or behavioral characters. Neotropical and Australian species came out as separate, apical clades. Asian species split between an apical branch, appearing as sister group to the neotropical clade, and basal taxa. New Guinean species were spread among several clades, suggesting a derivation from multiple origins. A well-supported clade includes the neotropical, Australian, and New Guinean species, with the southeast Asian N. takasagoensis and N. matangensis. It excludes the Asian species N. regularis, N. parvonasutus, and N. longinasus, which might deserve to be removed from Nasutitermes, as well as the long-legged Asian genera Hospitalitermes and Longipeditermes. A Gondwanan origin is proposed for the former clade, although an Old World origin of Nasutitermes followed by dispersal to Australia and South America cannot be excluded. PMID- 11020298 TI - Synthesis, antibacterial, and cytotoxic evaluation of certain 7-substituted norfloxacin derivatives. AB - We report herein the synthesis and biological evaluation of two series of 7 substituted norfloxacin derivatives. Most compounds tested in this study demonstrated better activity against methicillin-resistant Staphylococcus aureus than norfloxacin. Preliminary in vitro evaluation indicated that the 7-[4-(2 hydroxyiminoethyl)piperazin-1-yl] derivatives 3b-e possess distinct cytotoxicity profiles as compared with their alpha-methylene-gamma-butyrolactone counterparts, 4b,e: i.e., excellent activities against the renal cancer subpanel. Among them, 1 ethyl-6-fluoro-7-?4-[2-(4-chlorophenyl)-2-hydroxyiminoethyl]-1-p ipe razinyl?-4 oxo-1,4-dihydro-3-quinolinecarboxylic acid (3d) demonstrated the most significant activities against renal cancer cell lines, with log GI(50) values of -6.40 against CAK-1, -6.14 against RXF 393, and -7.54 against UO-31, compared with a mean log GI(50) value of -5.03. PMID- 11020300 TI - First data on the molecular phylogeography of scincid lizards of the genus Mabuya. AB - A 487-bp fragment of the mitochondrial 16S rRNA gene was sequenced in 26 species of the circumtropical lizard genus Mabuya and used to analyze phylogenetic relationships within the genus. The species from Africa and Madagascar formed a monophyletic group relative to the included Asian and South American taxa. The Malagasy species included (M. elegans, M. cf. dumasi, and M. comorensis) did not appear as a monophylum. Combined and separate analysis of the 16S data and additional sequences of the mitochondrial 12S rRNA, ND4, and cytochrome b genes (a total of 2255 bp) in one Asian, two Malagasy, and two African species also did not result consistently in a monophyletic grouping of the Malagasy taxa. However, a monophylum containing African and Malagasy taxa was strongly supported by the combined analysis. These preliminary results indicate that Mabuya probably colonized Madagascar from Africa through the Mozambique Channel. PMID- 11020302 TI - 28S rDNA evolution in the Eumalacostraca and the phylogenetic position of krill. AB - The Malacostraca are an ancient and morphologically diverse class of Crustacea. The phylogenetic position of one order within this class, the Euphausiacea ("krill," subclass Eumalacostraca) was investigated using 28S rDNA sequences from representatives of several malacostracan orders. Phylogenies for these sequences were estimated by maximum-likelihood and maximum-parsimony analysis. The results of these analyses produced a new scheme for evolution within the Eumalacostraca. The new phylogenies suggested that Euphausiacea are most closely related to the Mysida and not the Decapoda, as is generally thought. Furthermore, the Mysida were found not to be closely related to the Lophogastrida, which are often considered their sister taxon. These hypotheses were tested against the hypotheses of monophyly for the Eucarida, Mysidacea, and Peracarida and found to be significantly better on the basis of the 28S rDNA data. PMID- 11020301 TI - Phylogenetic investigations of the stephanoberyciformes and beryciformes, particularly whalefishes (Euteleostei: Cetomimidae), based on partial 12S rDNA and 16S rDNA sequences. AB - DNA data were collected from a number of acanthomorph fishes for 12S rDNA (30 sequences) and 16S rDNA (39 sequences) to investigate the phylogenetic relationships of genera within Cetomimidae (whalefishes) and of this family within the Stephanoberyciformes/Beryciformes assemblage. The Cetomimidae are apparently monophyletic. Within the family, species of Gyrinomimus and Cetomimus form a clade but the former genus is paraphyletic with respect to the latter. Cetostoma is sister to Ditropichthys rather than to Gyrinomimus plus Cetomimus as suggested by morphological analyses. Rondeletiidae + Cetomimidae + Barbourisiidae are shown, as expected from morphological analyses, as a monophyletic group in the 12S rDNA analyses, but not in the 16S rDNA or combined analyses, although the shortest trees showing the group require only one extra step in each case. These three families plus Melamphaidae (our sample of Stephanoberyciformes) are not shown as a group in any analysis, with Melamphaidae being sister to Berycidae in the 16S and combined analyses, but dispersed in the 12S analyses. Maximum parsimony trees without imposed constraints are notably shorter than trees constrained to show ordinal groupings or either of the two main current hypotheses of Stephanoberyciformes/Beryciformes relationships. The length difference is highly significant for most comparisons using either 12S or 16S rDNA sets or their combination, and significant or nearly so for all comparisons. In particular, the Beryciformes is unlikely to be monophyletic. The Holocentridae are included, with high bootstrap and Bremer support, in a clade of non beryciforms comprising the Gempylidae, Zeidae, and Atheriniformes (the only higher acanthomorphs sampled) and not with other Beryciform families. In these data, the Berycidae are the sister to the Melamphaidae, a stephanoberyciform family. PMID- 11020303 TI - Phylogenetic and biogeographic relationships of eastern Asian and eastern North American disjunct Suillus species (fungi) as inferred from nuclear ribosomal RNA ITS sequences. AB - Species of Suillus produce fleshy, pored mushrooms. They are important symbiotic (ectomycorrhizal) partners of many coniferous trees. The genus includes several putative eastern Asian and eastern North American disjunct species, i.e., the S. americanus-S. sibiricus and S. decipiens-S. spraguei complexes. Phylogenetic relationships among the groups were determined to further understand the biogeographic pattern. Analyses were based on 40 sequences of the ITS region of the nuclear ribosomal RNA tandem repeats, representing 18 distinct species/populations. Our phylogenetic analyses suggested that: (1) Chinese and United States' (U.S.) S. spraguei plus S. decipiens form a strongly supported monophyletic group, with North American S. decipiens and Chinese S. spraguei being sister taxa; (2) S. americanus, Asian and U.S. S. sibiricus, plus S. umbonatus form a clade supported by a high bootstrap value; and (3) little ITS sequence divergence exists within the latter group compared to the S. decipiens S. spraguei clade. Phylogenetic patterns revealed by this study imply a close phylogenetic relationship between eastern Asian and eastern North American disjunct population/species of Suillus. These fungi display relatively high host fidelity (at least to the host subgenus level), suggesting potential coevolutionary/comigratory trends. PMID- 11020304 TI - Phylogenetic relationships of north American field crickets inferred from mitochondrial DNA data. AB - A well-supported molecular phylogeny for North American Gryllus species based on a combined data set of mitochondrial (mt) DNA is presented. A total of 26 individuals representing 13 populations of 11 species of the genus Gryllus and 4 individuals of two outgroup species, Teleogryllus oceanicus and Acheta domestica, were sampled in this study. The complete cytochrome b gene (1036 bp) and a 500-bp fragment of the 16S rRNA gene were sequenced for each individual. Since results from separate analyses of the cytochrome b and 16S data sets, as well as a previously published mtDNA restriction-site data set, were not conflicting, all data were combined for phylogenetic analyses. The clade of European Gryllus was clearly separated from the North American clade. The amount of sequence divergence between these clades was significantly greater than within the clades, suggesting a basal drift-vicariant event in the genus. This is the first phylogenetic analysis of North American Gryllus that includes western species. Four well-supported groups were identified but their relationships showed no clear east-west structure. Our phylogeny supports the recent reassignment of G. integer Scudder 1901 from Texas to G. texensis Cade and Otte 2000. The evolution of cricket song and life cycle is discussed using the new phylogenetic framework. PMID- 11020305 TI - Slicing hyperdimensional oranges: the geometry of phylogenetic estimation. AB - A new view of phylogenetic estimation is presented where data sets, tree evolution models, and estimation methods are placed in a common geometric framework. Each of these objects is placed in a vector space where the character patterns are the basis vectors. This viewpoint allows intuitive understanding of various complex properties of the phylogeneticestimation problem structure. This is illustrated with examples discussing data set combinations, mixture models, consistency, and phylogenetic invariants. PMID- 11020306 TI - Mitochondrial gene fragments suggest paraphyly of the genus Panorpa (Mecoptera, Panorpidae). AB - Life history studies of scorpionfly species have been used to test predictions of evolutionary theory, but comparative analysis has been hampered by a lacking phylogeny of scorpionflies. We present a molecular phylogeny of selected panorpid scorpionflies inferred from mitochondrial 12S, 16S rRNA, and COI gene fragments, using parsimony and maximum-likelihood methods. Maximum-likelihood reconstructions depend on an explicit evolutionary substitution model; therefore, we estimated fit of substitution models to our data and used an optimal evolutionary substitution model in subsequent reconstructions. Both reconstruction methods converge on compatible trees with considerable statistical support for a majority of nodes. We performed parametric tests of most important phylogenetic conclusions employing the fitted GTR + %I + Gamma substitution model. Parametric bootstrapping allowed rejection of alternative explanations of the data set, where classical tests, like the KHY test, failed. Parametric bootstrapping confirmed that the association of Neopanorpa sp. with Asian Panorpa species is currently the superior explanation of the data set. Therefore, it is concluded that the genus Panorpa is most likely paraphyletic to the representative of the genus Neopanorpa. We conclude that the sequenced mitochondrial gene fragments appear to be well suited for future more comprehensive phylogenetic investigations of panorpid scorpionflies. PMID- 11020307 TI - Phylogeny and life history evolution of the genus Chrysoritis within the Aphnaeini (Lepidoptera: Lycaenidae), inferred from mitochondrial cytochrome oxidase I sequences. AB - Phylogenetic relationships among 26 South African species in the tribe Aphnaeini (Lepidoptera: Lycaenidae) were inferred from DNA characters of the mitochondrial gene cytochrome oxidase I (COI), using maximum-parsimony methods. The resulting phylogenetic estimate supports the systematic hypothesis made by Heath (1997, Metamorphosis, supplement 2), based on morphological characters, that at least three preexisting genera (Chrysoritis, Poecilmitis, and Oxychaeta) should be collapsed into the single monophyletic genus Chrysoritis. Two of the species groups described by Heath within Chrysoritis are also monophyletic, while one is paraphyletic and thus unsupported by the molecular data. Strong node support and skewed transition/transversion ratios suggest that two Chrysoritis clades contain synonymous species. Aphytophagy appears as a derived feeding strategy. Evolutionary patterns of ant association indicate lability at the level of ant genus, while association with different ant subfamilies may have played an ancestral and chemically mediated role in the diversification of South African aphnaeines. PMID- 11020308 TI - Homeobox gene diversification in the calcareous sponge, Sycon raphanus. AB - Knowledge of the developmental mechanisms in living basal metazoan phyla is crucial for understanding the genetic bases of morphological evolution in early animal history. We looked for homeobox genes in the calcareous sponge, Sycon raphanus, using the polymerase chain reaction. Partial sequences of eight homeoboxes were recovered, five of which are assignable to the NK-2 class of homeoboxes. The three remaining sequences are related members of a new class of homeoboxes, the Sycox class, showing limited similarity to bilaterian Lbx, Hlx, HEX, En, and Cad classes. Among the five NK-2 class homeoboxes are four closely related sequences occupying a divergent position within the class, the remaining one on the contrary showing high sequence similarity with members of the NK-2 family, a particular subgroup within the NK-2 class, previously known only from the Bilateria. This suggests that diversification of the NK-2 class occurred early in metazoan history. Altogether, the results reveal an unexpected diversification of homeobox genes in S. raphanus. PMID- 11020309 TI - A molecular framework for the phylogeny of the ant subfamily dolichoderinae. AB - Partial sequences are reported for the mitochondrial genes for cytochrome oxidase subunits 2 and 3 and for cytochrome b, and the entire sequence of the gene for tRNA(Leu)(UUR) for species from 14 genera of dolichoderine ants and from three outgroup genera. Considerable variation was observed between tRNA genes in the size of the TPsiC arm and the DHU and anticodon loops and whether or not the TPsiC stem possesses a GC pair. The outgroup taxa showed complete TAA CO1 stop codons, but dolichoderines have either TA or T. The outgroup taxa showed a noncoding gap between the CO1 and the tRNA(Leu)(UUR) genes. A phylogeny independent compatibility test using the amino acid sequences showed differences between the genes consistent with variation in evolutionary rates, according with other studies. Base compositions proved heterogeneous between species, hence phylogenetic analysis was restricted to the protein sequences using maximum likelihood and the mtREV24 replacement matrix. A maximum-likelihood consensus tree has similarities to those from morphological studies with some exceptions such Leptomyrmex falling within the dolichoderine genera rather than basally, and the accretion of genera formerly included under Iridomyrmex. Features of the tRNA genes and the CO1 termination codons agree quite well with the molecular phylogeny. PMID- 11020310 TI - Affinities among anostracan (Crustacea: Branchiopoda) families inferred from phylogenetic analyses of multiple gene sequences. AB - To gain insights into the relationships among anostracan families, molecular phylogenetic analyses were performed on nuclear (28S D1-D3 ribosomal DNA) and mitochondrial (16S rDNA, COI) gene regions for representatives of seven families and an outgroup. Data matrices used in the analyses included 951 base pairs (bp) of aligned sequences for 28S, 465 bp for 16S, and 658 bp (219 amino acids) for COI. Maximum-parsimony and maximum-likelihood methods were used to construct phylogenetic trees, enabling the evaluation of both previous hypotheses of taxonomic relationships among families based on morphology, and of the relative merits of independent versus simultaneous analyses of multiple data sets for phylogeny construction. Data from various combinations of the gene regions produced relatively congruent patterns of phylogenetic affinity. In most analyses, two monophyletic groups were resolved: one cluster included the families Polyartemiidae, Chirocephalidae, Branchinectidae, Streptocephalidae, and Thamnocephalidae, while the other contained the Artemiidae and Branchipodidae. Comparative analyses showed that combining gene regions in a single matrix generally resulted in increased resolution and support for each cluster relative to those obtained from single-gene analyses. Statistical tests demonstrated that morphology-based hypotheses of relationships among families had poorer support than those determined from molecular data, reflecting the homoplasy in characters used to differentiate families. PMID- 11020311 TI - The importance of being earnest: what, if anything, constitutes a "specimen examined?". PMID- 11020312 TI - Developments in immunoassay technology. PMID- 11020313 TI - Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays. AB - A very large proportion of modern immunoassays involve the use of synthetic solid phases to immobilize one of the reactants. These solid-phase immunoassays (SPIs) therefore involve ligand-receptor interactions that occur within a reaction volume close to the solution/solid phase interface. As a consequence, the immunochemistry/biochemistry of these ligand-receptor interactions differs from that of their counterparts in solution. Furthermore, the immobilization process can significantly alter the biological activity of the reactant; most adsorbed proteins on polystyrene or silicone are partially or largely denatured. Therefore the use of alternative methods of immobilization is attractive but may result in little increase in the amount of total functional reactant. However, all commonly used solid phases do not have the same properties or the same capacity for reactant immobilization or experience the same level of nonspecific binding. Empiricism plays a major role in SPIs. Derivations of mass law equations for measuring the antigen capture of solid-phase antibodies, for determining the affinity of solid phase for protein adsorption, and for estimating antibody affinity are reviewed. PMID- 11020314 TI - Expression immunoassay. AB - Expression immunoassay is a sensitive analytical method that takes advantage of coupled in vitro transcription and translation as a signal amplification technique. Essentially, the immunoassay is performed using a detection antibody that is labeled with an expressible fragment of DNA. The product of expression is a protein that can be used to generate a signal. Here we describe two distinct expression immunoassays; both are based on expression of DNA labels to produce active enzyme molecules which are subsequently detected through their enzymatic activities. The luciferase expression immunoassay uses a 2.1-kb DNA template as a reporter molecule. The DNA is attached to a biotinylated detection antibody via biotin-streptavidin linkage. After the detection antibody is immunoreacted with immobilized antigen and excess antibody is removed, the DNA label is expressed in vitro. A linear relationship exists between the bioluminescent signal, from luciferase activity, and the immobilized antigen. This expression immunoassay allows the detection of 5 x 10(4) antigen molecules. The second expression immunoassay makes use of the well-characterized alpha-complementation of beta galactosidase by employing an alpha-peptide encoding DNA as a reporter molecule. By monitoring the resulting beta-galactosidase activity with a fluorogenic substrate it was possible to detect as little as 3 fmol of immobilized antigen. Both expression immunoassays are amenable to automation and demonstrate the potential sensitivity that can be achieved using in vitro expression as a signal amplification method. PMID- 11020315 TI - Synthesis of bifunctional antibodies for immunoassays. AB - The synthesis of bifunctional antibodies using the principle of solid-phase synthesis is described. Two Fab' fragments were chemically linked together via a bismaleimide crosslinking reagent. The F(ab')(2) fragments from intact immunoglobulin G (IgG) were prepared using an immobilized pepsin column. Goat, mouse, and human antibodies were digested completely within 4 h. The F(ab')(2) fragments thus produced did not contain any IgG impurities. Fab' fragments were produced by reducing the heavy interchain disulfide bonds using 2 mercaptoethylamine. Use of the solid-phase reactor in the preparation of the bifunctional antibodies eliminated many of the time-consuming separation steps between the fragmentation and conjugation steps. This procedure facilitates the automation of bifunctional antibody preparation and the rapid optimization of reaction conditions. PMID- 11020316 TI - Site-specific photobiotinylation of antibodies, light chains, and immunoglobulin fragments. AB - The high affinity of biotin for avidin has been exploited for many antibody-based assays. This requires that biotin is covalently conjugated to the antibody molecule. Several chemically reactive biotinylation reagents are commercially available. Except for the attachment via sulfhydryl groups in the immunoglobulin (Ig) molecule, these reagents attach biotin randomly to various amino acid side chains. Although non-site-specific modification of antibodies does not interfere in most immunoassays, specific application and sensitive antibodies would benefit from site-specific biotinylation. Here we describe an affinity biotinylation technique based on a photoreactive biotin reagent. The design of this reaction was possible from the discovery of a conserved binding site in the variable Ig domain for nucleotides and nucleosides. The described photoaffinity biotinylation offers the advantages of ease, convenience, and production of a reproducible and defined biotinylated antibody preparation. PMID- 11020317 TI - Monoclonal antibody-based enzyme immunoassay for mercury(II) determination. AB - A monoclonal antibody (K3C6) was developed against Hg(II) and applied in different enzyme immunoassay (EIA) formats to determine the test system with the highest sensitivity. A detection limit of 1. 0 microg/L Hg(II) could be achieved with a competitive format in contrast to a detection limit of 2.1 microg/L Hg(II) with a noncompetitive EIA. A competitive displacement EIA yielded the best detection limit of 0.4 microg/L Hg(II) and was well suited to measuring real samples. For this purpose different water samples were diluted at least 1:10 to avoid matrix effects and subsequently spiked with 1 microg/L HgCl(2). Recovery of the spiked samples was between 80 and 120%. PMID- 11020318 TI - Development of rapid one-step immunochromatographic assay. AB - An analytical system for a one-step immunoassay has been constructed using the concept of immunochromatography. The system employed two different antibodies that bound distinct epitopes of an analyte molecule: an antibody labeled with a signal generator (e.g., colloidal gold), which was placed in the dry state at a predetermined site on a glass-fiber membrane, and another antibody immobilized on the surface of a nitrocellulose membrane. Three membranes, one with the tracer, one with immobilized antibody, and a cellulose membrane as the absorbent of medium (in a sequence from the bottom), were attached to a plastic film and cut into strips. Aqueous medium containing analyte absorbed from the bottom end of the immunostrip dissolved the labeled antibody, and the antigen-antibody binding complex formed was transported into the next nitrocellulose membrane by the flow caused by capillary action. The complex subsequently reacted with the immobilized antibody, which generated a signal in proportion to the analyte concentration. The convective mass transfer of the immunoreactant to the binding partner allowed the assay to be performed with no handling of reagents. The reaction, however, was carried out under nonequilibrium conditions, which resulted in decreased sensitivity as compared with assays performed in an equilibrium mode (e.g., ELISA). To minimize such sacrifice, major factors that control system performance were identified and the system was then devised under optimal conditions. PMID- 11020319 TI - Fluorescence polarization competition immunoassay for tyrosine kinases. AB - To increase the sensitivity and throughput of protein tyrosine kinase (PTK), simple, homogeneous, nonradioactive, direct and indirect fluorescence polarization (FP) protein tyrosine kinase immunoassays have been developed that are compatible with high-throughput and ultrahigh-throughput screening for developing drugs. In the direct method, a fluorescinylated peptide substrate is incubated with the kinase, ATP, and antiphosphotyrosine antibody. The phosphorylated peptide product is immunocomplexed with the antiphosphotyrosine antibody, resulting in an increase in the polarization signal. Since the direct method can be used only with a peptide substrate and requires large amounts of antiphosphotyrosine antibody, a modified indirect method, wherein a phosphorylated peptide or protein produced by kinase reaction will compete with a fluorescent phosphopeptide used as a tracer for immunocomplex formation with phosphotyrosine antibody, was developed. In this format kinase activity will result in loss of the polarization signal. Both the direct and indirect FP-PTK immunoassays have been compared with a more commonly used (32)PO(4) transfer assay and validated using lymphoid T-cell protein tyrosine kinase (Lck). In both assays, Lck activity showed a similar dependence on ATP, Lck enzyme, and peptide substrate concentration, comparable to the (32)PO(4) transfer assay. Inhibition by staurosporine and the Lck inhibitor 4-amino-5-(methylphenyl)-7-(tert butyl)pyrazolo[3, 4-d]pyrimidine in these two FP assays was similar to that obtained in the (32)PO(4) transfer assay. The advantages of these FP-PTK assays over the other kinase assays, besides high sensitivity, are use of inexpensive nonisotropic substrate; environmental safety; homogeneous nature of FP kinase assays that are done in the same tube (or in a well of 96- or 384-well microtiter plates), without separation, precipitation, or washing; and increase of throughput. PMID- 11020320 TI - Fluorescence polarization immunoassay: detection of antibody to Brucella abortus. AB - Fluorescence polarization immunoassay (FPA) is a homogeneous immunoassay useful for rapid and accurate detection of antibody or antigen. The principle of the assay is that a fluorescent dye (attached to an antigen or an antibody fragment) can be excited by plane-polarized light at the appropriate wavelength. As a rule, a small molecule rotates faster when in solution than a larger molecule. The rotation rate may be assessed by measuring light intensity in the vertical and horizontal planes. Generally, the time it takes a molecule to rotate through a given angle is an indication of its size. When a small molecule that rotates rapidly is bound to a larger molecule, the rotation rate is decreased and this decrease is measured. Because it is a primary antigen-antibody interaction, the rate of reaction is very rapid and usually a result may be obtained in minutes. This technology was applied to the detection of antibody to Brucella abortus in serum and milk, providing for the first time a rapid primary binding assay that is cost effective for use in the field. PMID- 11020321 TI - Surface plasmon resonance-based immunoassays. AB - Surface plasmon resonance (SPR) has been successfully incorporated into an immunosensor format for the simple, rapid, and nonlabeled assay of various biochemical analytes. Proteins, complex conjugates, toxins, allergens, drugs, and pesticides can be determined directly using either natural antibodies or synthetic receptors with high sensitivity and selectivity as the sensing element. Immunosensors are capable of real-time monitoring of the antigen-antibody reaction. A wide range of molecules can be detected with lower limits ranging between 10(-9) and 10(-13) mol/L. Several successful commercial developments of SPR immunosensors are available and their web pages are rich in technical information. This review highlights many recent developments in SPR-based immunoassay, functionalizations of the gold surface, novel receptors in molecular recognition, and advanced techniques for sensitivity enhancement. Furthermore, it describes the challenge of current problems and provides some insights toward the future technologies. PMID- 11020322 TI - Application of imprinted synthetic polymers in binding assay development. AB - The first part of the review describes a method for the synthesis of molecularly imprinted polymers for use in binding assays. The method considers the many factors involved that affect the recognition properties of the materials and describes an approach to screening and optimization of these factors. The second part describes the development of binding assays using such polymers. This includes the use of different labels, the effect of solvent and buffer, the scale of the assay (amount of solid polymer), and how these influence the quality of the assay in terms of sensitivity, selectivity, and speed of analysis. PMID- 11020324 TI - Editorial PMID- 11020323 TI - In vitro selected oligonucleotides as receptors in binding assays. AB - This paper describes a new binding assay that uses oligodeoxyribonucleotides (DNAs) obtained by the in vitro selection method instead of antibodies. DNAs that specifically bound to a target molecule were selected, labeled with some probes, and then used to detect the target molecule by staining or competitive binding assay. PMID- 11020325 TI - Mutagenesis assays in yeast. AB - Analyzing mutation spectra is a very powerful method to determine the effects of various types of DNA damage and to understand the workings of various DNA repair pathways. However, compiling sequence-specific mutation spectra is laborious; even with modern sequencing technology, it is rare to obtain spectra with more than several hundred data points. Two assay systems are described for yeast, one for insertion/deletion mutations and one for base substitution mutations, that allow determination of specific mutations without the necessity of DNA sequencing. The assay for insertion/deletion mutations uses a variety of different simple repeats placed in frame with URA3 such that insertions or deletions lead to a selectable Ura(-) phenotype; essentially all such mutations are in the simple repeat sequence. The assay for base substitution mutations uses a series of six strains with different mutations in one essential codon of the CYC1 gene. Because only true reversions lead to a selectable phenotype, the bases mutated in any reversion event are known. The advantage of these assays is that they can quantitatively determine over several orders of magnitude the types of mutations that occur under a given set of conditions, without DNA sequencing. PMID- 11020326 TI - In vivo technique for determining transcriptional mutagenesis. AB - When an elongating RNA polymerase encounters DNA damage on the template strand of a transcribed gene it can either be arrested by or be transcribed through the lesion. Lesions that arrest RNA polymerases are thought to be subject to transcription-coupled repair, whereas that damage that is bypassed can cause miscoding, resulting in "mutations" in the transcript (transcriptional mutagenesis). We have developed a technique using a plasmid-based luciferase reporter assay to determine the extent to which a particular type of DNA base modification is capable of causing transcriptional mutagenesis in vivo. The system uses Escherichia coli strains with different DNA repair backgrounds and is designed to detect phenotypic changes caused by transcriptional mutagenesis under nongrowth conditions. In addition, this method is capable of indicating the extent to which a particular DNA repair enzyme (or pathway) suppresses the occurrence of transcriptional mutagenesis. Thus, this technique provides a tool with which the effects of various genes on non-replication-dependent pathways resulting in the generation of mutant proteins can be gauged. PMID- 11020327 TI - Measurement of oxidative DNA damage and repair in specific DNA sequences. AB - We describe two methods that were developed in our laboratory to measure the production and repair of oxidative DNA damage in specific DNA sequences. Both of these methods rely on the use of monoclonal antibodies against modified nucleotides to separate DNA sequences that contain damage from those in which repair has occurred. In one case, the modified base is bromodeoxyuridine, which is inserted into the DNA during the repair synthesis step of excision repair. An antibody against this modified base is used to detect the production of the bromodeoxyuridine in the repair patch. This approach allows for the measurement of repair of any DNA lesion whose removal is accompanied by the production of a DNA repair patch. In the other case, the modified base is thymine glycol, an oxidized base that is produced by hydrogen peroxide and ionizing radiation. Using a monoclonal antibody that recognizes this altered base, detection of the production and repair of a specific base damage in a DNA sequence can be accomplished. These approaches are used in our laboratory to examine the transcription-coupled repair of oxidative DNA damage in both yeast and mammalian cells. PMID- 11020328 TI - Analysis of gene-specific DNA damage and repair using quantitative polymerase chain reaction. AB - Soon after discovery of the polymerase chain reaction (PCR), various laboratories have attempted to use quantitative PCR (QPCR) to detect DNA damage in specific gene segments. The development of techniques that facilitate long PCR increased the sensitivity of the assay so that biologically relevant doses of DNA-damaging agents could be assessed. QPCR has been used to survey DNA damage induced by different genotoxicants and to establish the repair kinetics of numerous genes. Current work seeks to analyze damage and repair in specific genes from animals exposed to specific DNA-damaging agents such as oxidative stress. PMID- 11020329 TI - Mapping oxidative DNA damage using ligation-mediated polymerase chain reaction technology. AB - Reactive oxygen species induce a pharmacopoeia of oxidized bases in DNA. DNA can be cleaved at most of the sites of these modified bases by digestion with a combination of two base excision repair glycosylases from Escherichia coli, Fpg glycosylase, and endonuclease III. The frequency of the resulting glycosylase dependent 5'-phosphoryl ends can be mapped at nucleotide resolution along a sequencing gel autoradiogram by a genomic sequencing technique, ligation-mediated polymerase chain reaction (LMPCR). In cultured rat cells, the frequency of endogenous oxidized bases in mitochondrial DNA is sufficiently high, about one oxidized base per 100 kb, to be directly mapped from 0.1 microg of total cellular DNA preparations by LMPCR. Nuclear DNA has a lower frequency of endogenous oxidative base damage which cannot be mapped from 1-microg preparations of total cellular DNA. Preparative gel electrophoresis of the PGK1 and p53 genes from 300 microg of restriction endonuclease-digested genomic DNA showed a 25-fold enrichment for the genes and, after endonuclease digestion followed by LMPCR, gave sufficient signal to map the frequency of oxidized bases from human cells treated with 50 microM H2O2. PMID- 11020330 TI - Measuring DNA damage using capillary electrophoresis with laser-induced fluorescence detection. AB - Damage to cellular DNA is implicated in the early stages of carcinogenesis and in the cytotoxicity of many anticancer agents, including ionizing radiation. Sensitive techniques are required for measuring cellular levels of DNA damage. We describe in detail a novel immunoassay that makes use of the resolving power of capillary electrophoresis and the sensitivity of laser-induced fluorescence detection. An example is given of the detection of thymine glycol in DNA produced by irradiation of human cells with a clinical dose of 2 Gy. A detection limit of approximately 10(-21) mol allowed us to monitor the repair of the lesion and to suggest that the cellular repair response may be inducible. PMID- 11020331 TI - Detection of abasic sites and oxidative DNA base damage using an ELISA-like assay. AB - Reactive oxygen species produce a wide spectrum of DNA damage, including oxidative base damage and abasic (AP) sites. Many procedures are available for the quantification and detection of base damage and AP sites. However, either these procedures are laborious or the starting materials are difficult to obtain. A biotinylated aldehyde-specific reagent, ARP, has been shown to react specifically with the aldehyde group present in AP sites, resulting in biotin tagged AP sites in DNA. The biotin-tagged AP sites can then be determined colorimetrically with an ELISA-like assay, using avidin/biotin-conjugated horseradish peroxidase as the indicator enzyme. The ARP assay is thus a simple, rapid, and sensitive method for the detection of AP sites in DNA. Furthermore, removal of damaged base by DNA N-glycosylases generates AP sites that can be measured by the ARP reagent. By coupling the ARP assay with either endonuclease III from Escherichia coli or 8-oxoguanine N-glycosylase (OGG1) from yeast, investigators can rapidly determine the amount of oxidative pyrimidine damage (endonuclease III-sensitive sites) or purine damage (OGG1-sensitive sites) in cellular DNA, respectively. An increased level of oxidative damage has been implicated in several age-related human diseases such as Alzheimer's disease, amyotrophic lateral sclerosis, and Parkinson's disease, as well as the aging process. The sensitivity and simplicity of the ARP assay thus make it a valuable method for investigators who are interested in estimating the level of oxidative DNA damage in cells and tissues derived from patients with various age-related diseases or cancers. PMID- 11020332 TI - Low- and high-resolution mapping of DNA damage at specific sites. AB - Measurement of DNA damage and repair at the nucleotide level in intact cells has provided compelling evidence for the molecular details of these events as they occur in intact organisms. Furthermore, these measurements give the most accurate picture of the rates of repair in different structural domains of DNA in chromatin. In this report, we describe two methods currently used in our laboratories to map DNA lesions at (or near) nucleotide resolution in yeast cells. The low-resolution method couples damage-specific strand breaks in DNA with indirect end-labeling to measure DNA lesions over a span of 1.5 to 2 kb of DNA sequence. The resolution of this method is limited by the resolution of DNA length measurements on alkaline agarose gels (about +/-20 bp on average). The high-resolution method uses streptavidin magnetic beads and special biotinylated oligonucleotides to facilitate end-labeling of DNA fragments specifically cleaved at damage sites. The latter method maps DNA damage sites at nucleotide resolution over a shorter distance (<500 bp), and is constrained to the length of DNA resolvable on DNA sequencing gels. These methods are used in tandem for answering questions regarding DNA damage and repair in different chromatin domains and states of gene expression. PMID- 11020333 TI - Identification, characterization, and purification of DNA glycosylase/AP lyases by reductive crosslinking to 2'-deoxyribooligonucleotides containing specific base lesions. AB - This paper describes a reductive amination crosslinking protocol that facilitates identification and characterization of a class of DNA repair enzymes, DNA glycosylase/AP lyases, which are involved in base excision repair. This crosslinking technique has been used to identify enzymes in crude extracts and in partially purified enzyme preparations, to isolate proteins for sequencing, and to confirm the reaction mechanism of members of this enzyme family. Chemical reduction of the Schiff's base enzyme-substrate intermediate to a stable amine results in the formation of an irreversible covalent bond between the substrate lesion situated within a 2'-deoxyoligonucleotide and the repair enzyme. This complex can be detected by gel electrophoresis and can also be isolated and analyzed by amino acid sequencing. PMID- 11020334 TI - Nucleotide excision repair in oocyte nuclear extracts from Xenopus laevis. AB - We have developed efficient DNA repair extracts derived from the unusually large nuclei of Xenopus oocytes. These extracts use nucleotide excision repair (NER) to completely remove bulky adducts from DNA. There is very little or no synthesis on control, undamaged DNA, indicating the extracts do not have significant nonspecific nuclease activity, and repair of cyclobutane pyrimidine dimers (CPDs) occurs in the dark, indicating that NER, and not photolyase, is responsible for CPD repair. The extracts can be inactivated with antibodies specific to repair proteins and then repair activity can be restored by adding purified recombinant protein. Here we describe detailed protocols for preparing Xenopus nuclear repair extracts. PMID- 11020336 TI - Application of chemically induced dynamic nuclear polarization to the nitration of N-acetyltyrosine and to some reactions of peroxynitrite. AB - By the observation of chemically induced dynamic nuclear polarization in (15)N NMR spectroscopy it has been shown that nitration of N-acetyltyrosine, even under acidic conditions, is largely a radical process. In the alkaline reaction of tyrosine with peroxynitrite the main products are nitrite and nitrate, both produced by a radical pathway, and tyrosine nitration is a minor reaction. It is suggested that tyrosine catalyzes the production of NO(*)(2) and HO(*) from peroxynitrite. PMID- 11020335 TI - Differential and constitutive expression of neuronal, inducible, and endothelial nitric oxide synthase mRNAs and proteins in pathologically normal human tissues. AB - Nitric oxide (NO) is produced by NO synthases (nNOS, iNOS, and eNOS) expressed in various human tissues and depending on the amount of NO produced in each tissue, the physiological function of NO is determined. However, due to the difficulty in obtaining normal human tissues, little is known about the basal levels of each of the three NOS mRNAsand proteins expressed constitutively in various human tissues. Results of the present study indicate that the basal levels of each of the three NOS mRNAs and proteins expressed in various regions of brain and peripheral tissues are different both in their sizes and in their contents. In Northern blot analysis, two different-sized mRNAs were found for each NOS isozymes: for the nNOS, approximately 12 and <12 kb mRNAs; for the iNOS, 4.2 and 4.5 kb mRNAs; for the eNOS, 4.2 and 4.4 kb mRNAs. In the Western blot, several different-sized NOS proteins were detected ( approximately 160, approximately 140, and approximately 130 kDa for nNOS; approximately 130 kDa for iNOS and eNOS) with tissue-specific expression patterns. These differential expression patterns of NOS mRNAs and proteins were caused by alternative splicing in the open-reading frame, and 5'- and/or 3'-untranslated regions of NOS mRNAs. These results suggest that regulation for differential expression of the three NOS genes in various human tissues may occur by alternative splicing of the NOS mRNAs in tissue specific patterns. PMID- 11020337 TI - Distinct subcellular localization and mRNA expression of neuronal nitric oxide synthase in the nucleus dorsalis and red nucleus and their correlation with inducible transcription factors after spinal cord hemisection. AB - We previously reported on the differential expression of neuronal nitric oxide synthase (nNOS) in neurons of the nucleus dorsalis (ND) and red nucleus (RN), as well as differential roles of nitric oxide (NO) in these two distinct groups' neurons characterized with different nNOS phenotypes after lower thoracic spinal cord hemisection. To further understand the enzyme, nNOS expression was studied at the subcellular and mRNA levels by using electron microscopic immunohistochemistry (EM-IHC) and in situ hybridization respectively. Possible transcriptional regulation by c-Jun or CREB in the differential nNOS expression in both ND and RN neurons was also studied. nNOS mRNA was not found in the normal ND neurons, but was shown in the normal RN neurons. After spinal cord hemisection, nNOS mRNA was induced in the ipsilateral ND, while upregulated on both sides of the RN, which preceded protein induction or upregulation. By EM IHC, nNOS immunoreaction products were predominantly bound to the membrane of the mitochondria, rough endoplasmic reticulum (rER), Golgi apparatus, and nuclear envelope in the RN neurons of normal rats as well as rats subjected to spinal cord hemisection. In contrast, nNOS-immunoreactive deposits in the experimental ND neurons were found to be mainly granular, being dispersed throughout the cytoplasmic matrix. It is speculated that the differential subcellular localizationof nNOS indicates that axotomy may trigger different nNOS transcripts and lead to different nNOS isoform expression in the normally non-nNOS- and normally nNOS-containing neurons. c-Jun was induced in the ipsilateral ND neuronsand upregulated only in the contralateral RN neurons. Activation of CREB by phosphorylation was occasionally detectable in the ND neurons, but not in the RN neurons. Double-labeling data showed a large proportion of c-Jun and nNOS colocalization in neurons of the ipsilateral ND and contralateral RN after spinal cord hemisection. However, dissociation of nNOS expression kinetics with c-Jun was observed in the ipsilateral RN. The results implied that nNOS expression might not be under the direct transcriptional regulation by c-Jun, although it seemed to be closely related to the c-Jun expression. PMID- 11020338 TI - Depletion of intracellular glutathione reduces mutations by nitric oxide-donating drugs. AB - The Mutatect system is a mouse tumor line in which mutations at the hypoxanthine phosphoribosyltransferase (Hprt) locus can be readily detected both in vitro and in vivo. We have previously shown that the nitric oxide-generating drugs, glyceryl trinitrate (GTN) and sodium nitroprusside (SNP), can induce mutations that are readily detected in these cells. In the present report, we have tested the effect of glutathione depletion by buthionine sulfoximine (BSO) on cytotoxicity and mutagenicity by these two drugs. Exposure for 24 h to either drug (123 microM GTN; 500 microM SNP) induced mutations with relatively little cytotoxicity. Pretreatment with 50 microM BSO for 24 h, and then removal at the time of GTN or SNP addition, enhanced cytotoxicity to a modest extent. However, mutagenicity induced by both GTN and SNP was largely abolished. BSO did not affect nitrite accumulation in the medium over a 24-h period, indicating no inhibition of bioactivation of GTN or SNP. Maintaining BSO in the medium for 24 h prior and throughout the period of exposure to GTN or SNP produced a similar effect on mutations. N-Acetylcysteine and oxothiazolidine-4-carboxylate, drugs that are used to increase intracellular glutathione, also blocked mutations. We postulate that a product of the reaction between nitric oxide and intracellular glutathione, such as GSNO or some species derived from it, is promutagenic. PMID- 11020339 TI - Effect of curcumin on the production of nitric oxide by cultured rat mammary gland. AB - We have hypothesized that one aspect of the antitumor activity of curcumin (diferuloylmethane) during the promotion stage of mammary gland tumorigenesis may be linked to reduction of free radicals (Inano et al., Carcinogenesis, 20: 1011 1018, 1999). Nitric oxide (NO) has been found to inflict damage on important biomolecules, and the overproduction of NO in diseases may be implicated in carcinogenesis and tumor progression. We have reported that the presence of three isoforms of nitric oxide synthases (NOS) and NO generation in the mammary gland correlate with the mammary gland development and mammary carcinogenesis. We, therefore, investigated the inhibitory activity of curcumin for the production of NO in rat mammary glands by using an organ culture system to validate the effectiveness and usefulness of curcumin in the pathophysiology of the mammary gland. A diced mammary gland (approximately 3 mm cubes) from the inguinal part of a female Wistar-MS rat treated with estradiol and progesterone was cultured with 2 ml of 5% FCS/DMEM in the presence or absence of LPS (0.5 microg/ml) for 2-3 days. Curcumin ( approximately 100 microM) was added at the same time to the LPS treated cultures. In some experiments, curcumin was added to the culture after the LPS had been washed out. The NO production was significantly increased (by almost 20-fold compared to the control) by the addition of LPS to the culture system. This enhancement of NO production by LPS was reduced to 76 and to 56% by addition of 30 and 100 microM curcumin, respectively, to the culture. When LPS was eliminated from the culture after prestimulation for 1 day, the production of NO by the mammary gland dropped off, although some NO was still detectable. Curcumin did not further inhibit the production of NO by the prestimulated mammary gland after the elimination of LPS from the culture. The inducible nitric oxide synthase (iNOS, 122 kDa) and endothelial nitric oxide synthase (eNOS, 152 kDa) isoforms were detected in the mammary gland extracts at the end of the organ culture. The quantity of iNOS was apparently increased in the gland treated with LPS, while the eNOS expression was clearly diminished. Curcumin (100 microM) obviously suppressed the iNOS expression in the mammary glands cultured with LPS, and a recovery in the eNOS expression was observed. On the other hand, curcumin exhibited scavenging activity for the NO released from N-ethyl-2-(1-ethyl-2 hydroxy-2-nitrosohydrazino)-ethanamine (NOC 12), a NO donor compound, in the coincubation mixture. These results indicate that curcumin has the ability to inhibit iNOS induction by LPS in the mammary gland and to scavenge NO radicals, which might explain, at least partly, its therapeutic properties in inflammation of the mammary gland. PMID- 11020340 TI - NO(x) contamination in laboratory ware and effect of countermeasures. AB - Contamination of various types of laboratory wares with NO(x) (NO(-)(2) and NO( )(3)) was assessed systematically and the effect of extensive washing as a countermeasure was evaluated. Mean NO(x) contamination arising from a model procedure for NO(x) determination in plasma was 0.93 microM (range, 0.35-1.49 microM). The major source of contamination included conical tubes (54.8%) and pipette tips used for transfer of solution (12.3-16.3%). Except for soft glassware, most NO(x) contamination could be washed out by pure water. Although NO(x) contamination in respective laboratory wares could be reduced below detection levels by extensive washing, summation of the contamination through the model procedure could not be completely abolished (but the effect of washing persisted at least 10 days). Heavy contamination was noted in glassware (especially soft glass) and ultrafiltration units, which was difficult to remove. Several types of vacuum blood sampling tubes contained various levels of NO(x). Our results indicated that a small but significant amount of contamination remained in laboratory ware even after extensive washing, and that it is advisable to avoid the use of glassware (soft glass), ultrafiltration units, and vacuum blood sampling tubes during the processing of clinical sampling for the measurement of NO(x). PMID- 11020341 TI - Nitrosylation of manganese(II) tetrakis(N-ethylpyridinium-2-yl)porphyrin: a simple and sensitive spectrophotometric assay for nitric oxide. AB - Reaction between NO(*) and manganese tetrakis(N-ethylpyridinium-2-yl)porphyrin (Mn(III)TE-2-PyP(5+)) was investigated at 25 degrees C. At high excess of NO(*) (1.5 mM) the reaction with the oxidized, air-stable form Mn(III)TE-2-PyP(5+) (5 microM), proceeds very slowly (t(1/2) congruent with 60 min). The presence of excess ascorbate (1 mM) produces the reduced form, Mn(II)TE-2-PyP(4+), which reacts with NO(*) stoichiometrically and in the time of mixing (k congruent with 1 x 10(6) M(-1) s(-1)). The high rate of formation and the stability of the product, Mn(II)TE-2-PyP(NO)(4+) (?Mn(NO)?(6)), make the reaction outcompete the reaction of NO(*) with O(2). Our in vitro measurements show a linear absorbance response upon addition of NO to a PBS, pH 7.4, solution containing an excess of ascorbate over Mn(III)TE-2-PyP(5+). Thus, the observed interactions can be the basis of a convenient and sensitive spectrophotometric assay for NO(*). Also, it may have important implications for the in vivo behavior of Mn(III)TE-2-PyP(5+) which is currently exploited as a possible therapeutic agent for various oxygen radical related disorders. PMID- 11020342 TI - Overexpression of neutrophil neuronal nitric oxide synthase in Parkinson's disease. AB - Much evidence supports a role of nitric oxide (.NO) and peroxynitrite (ONOO(-)) in experimental and idiopathic Parkinson's disease (PD); moreover, an overexpression of neuronal nitric oxide synthase (nNOS) was recently reported in the basal ganglia of PD patients. In accord, we previously found a 50% increased.NO production rate during the respiratory burst of circulating neutrophils (PMN) from PD patients. As PMN express the nNOS isoform, the objective of the present study was to ascertain whether this increased.NO production is representative of nNOS gene upregulation. PMN were isolated from blood samples obtained from seven PD patients and seven age- and sex-matched healthy donors; nNOS mRNA was amplified by reverse transcriptase-polymerase chain reaction and the products were hybridized with a probe for nNOS. Nitrotyrosine containing proteins and nNOS were detected by Western blot and NO production rate was measured spectrophotometrically by the conversion of oxymyoglobin to metmyoglobin. The results showed that both.NO production and protein tyrosine nitration were significantly increased in PMN isolated from PD patients (PD 0.09 +/- 0.01 vs 0.06 +/- 0.008 nmol min(-1) 10(6) cells(-1); P < 0.05). In addition, five of the seven PD patients showed about 10-fold nNOS mRNA overexpression; while two of the seven PD patients showed an expression level similar to that of the controls; detection of nNOS protein was more evident in the former group. In summary, it is likely that overexpression of nNOS and formation of ONOO(-) in PMN cells from PD patients emphasizes a potential causal role of.NO in the physiopathology of the illness. PMID- 11020343 TI - AIDS: are we doing enough? PMID- 11020344 TI - In the beginning: induction of prenatal tolerance still an elusive goal. PMID- 11020345 TI - Development of biomaterials for gene therapy. AB - Novel biocompatible polymeric gene carriers have been examined for their potential in treating various genetic and acquired diseases. The use of polymeric gene carriers may overcome the current problems associated with viral vectors in safety, immunogenicity, and mutagenesis. However, effective polymer-based gene therapy requires the control of cellular access and uptake, intracellular trafficking, and nuclear retention of plasmid DNA. Inefficient endosomal release, cytoplasmic transport, and nuclear entry of plasmids are currently limiting factors in the use of polymers for effective plasmid-based gene therapy. Therefore, several different polymeric gene carriers have been designed recently in an attempt to overcome these problems. This review explores the conceptual and experimental aspects of polymer-based gene delivery and presents an overview on the recent use of polymers to enhance the effectiveness of plasmid-based systems. Despite their current limitations, polymeric carriers have significant potential as commercially viable gene medicines. PMID- 11020346 TI - Inhibition of experimental lung metastasis by aerosol delivery of PEI-p53 complexes. AB - Mutations in the p53 tumor suppressor gene and the pathways mediated by the p53 protein are common in many human cancers. Replacement of functional p53 by gene therapy is a potential way of combating these cancers and the associated drug resistance and tumor growth. Aerosol delivery of genes is a noninvasive way of targeting genes to the lung for gene therapy. Here we demonstrate, using a murine melanoma lung metastasis model, that aerosol delivery of polyethyleneimine-p53 (PEI-p53) complexes inhibits the growth of lung metastasis. A significantly reduced number of visible foci were observed in C57BL/6 mice injected with B16 F10 melanoma and treated with PEI-p53 complexes by aerosol for 3 weeks at twice a week. Fifty percent of the mice in the PEI-p53-treated group exhibited no visible tumor foci. There was a significant reduction in the lung weights of p53-treated mice (P < 0.01) compared to control groups. The tumor burden was also significantly lower (P < 0.001) in mice treated with PEI-p53 complexes. No extrapulmonary metastasis was observed in the groups treated with PEI-p53 complexes compared to 50% of the mice in control groups, which showed metastasis to lymph nodes in the neck or abdomen. Treatment with PEI-p53 aerosol also led to about a 50% increase in the mean length of survival of the mice injected with B16 F10 cells. These data suggest that delivery of the p53 gene by aerosol using PEI as the gene delivery vector can inhibit the growth of lung metastasis. PMID- 11020347 TI - Tumor growth inhibition by intratumoral inoculation of defective herpes simplex virus vectors expressing granulocyte-macrophage colony-stimulating factor. AB - To evaluate the potential of defective herpes simplex virus (HSV) amplicon vectors as in vivo cytokine gene transfer vehicles for active immunotherapy, we generated a defective HSV vector that encodes the murine granulocyte-macrophage colony-stimulating factor (GM-CSF) gene, using a replication-defective HSV as helper virus. A variety of murine tumor cell lines were efficiently infected in vitro with the defective GM-CSF vector (dvGM), and this led to the synthesis and secretion of murine GM-CSF. In an established bilateral subcutaneous tumor model with Harding-Passey murine melanoma, unilateral intratumoral inoculation of dvGM significantly inhibited tumor growth of both the inoculated and noninoculated contralateral tumors. This tumor inhibition was dose-dependent and resulted in increased survival of the dvGM-treated mice. Inoculation of a lacZ-expressing defective vector had no effect on tumor growth. We conclude that this defective HSV vector system offers an effective method for cytokine gene delivery in vivo and that GM-CSF expression in tumors has antitumor activity. PMID- 11020348 TI - Strong promoters are the key to highly efficient, noninflammatory and noncytotoxic adenoviral-mediated transgene delivery into the brain in vivo. AB - Using the major immediate early murine cytomegalovirus (MIEmCMV) promoter to drive expression of beta-galactosidase, we have demonstrated that, following adenoviral-mediated transduction of brain cells in vivo, a single viral infectious unit is capable of producing detectable levels of transgene expression and that gene transfer into the brain is close to 100% efficient. By reducing 100 fold the amount of virus needed to detect large numbers of transduced brain cells, we were able to completely eliminate the cellular inflammation and viral cytotoxicity associated with the delivery of adenoviral vectors into the brain compared to saline-injected controls. These results demonstrate that a strong promoter is necessary to allow the use of low concentrations of adenoviral vectors for gene transfer into the brain, thereby eliminating deleterious side effects and increasing the potential efficacy of gene therapy. PMID- 11020349 TI - Characterization of a class of cationic peptides able to facilitate efficient protein transduction in vitro and in vivo. AB - Protein transduction domains (PTDs), such as the third helix of the Drosophila Antennapedia homeobox gene (Antp) and the HIV TAT PTD, possess a characteristic positive charge on the basis of their enrichment for arginine and lysine residues. To determine whether cationic peptides are able to function as protein transduction domains, 12-mer peptide sequences from an M13 phage library were selected for synthesis on the basis of their varying cationic charge content. In addition, polylysine and polyarginine peptides were synthesized in order to assess the effect of charge contribution in protein transduction. Coupling of the biotinylated peptides to avidin-beta-galactosidase facilitated transduction in a wide variety of cell lines and primary cells, including islet beta-cells, synovial cells, polarized airway epithelial cells, dendritic cells, myoblasts, and tumor cells. Two of the peptides, PTD-4 and PTD-5, mediated transduction nearly 600-fold more efficiently than a random control peptide, but with an efficiency similar to the TAT PTD and the 12 mers of polylysine and polyarginine. Furthermore, confocal analysis of biotinylated peptide-streptavidin-Cy3 conjugates demonstrated that the internalized PTDs are found in both the nuclei and the cytoplasm of treated cells. When tested in vivo, the PTDs were able to facilitate efficient and rapid protein delivery into rabbit synovium and mouse solid tumors following intraarticular and intratumoral administration, respectively. These novel PTDs can be used to transfer therapeutic proteins and DNA for the treatment of a wide variety of diseases, including arthritis and cancer. PMID- 11020350 TI - Intracellular Fas ligand expression causes Fas-mediated apoptosis in human prostate cancer cells resistant to monoclonal antibody-induced apoptosis. AB - Several laboratories have attempted with little success to induce Fas-mediated apoptosis in prostate cancer (PCa) cells, using different external Fas agonists, i.e., anti-Fas antibodies and membrane-bound FasL. The present study confirms these earlier results using the anti-Fas antibody CH-11 in five human PCa cell lines (PPC-1, LNCaP, PC-3, TSU-Pr1, and DU145). However, intracellular murine FasL expression induced Fas-mediated apoptosis in all CH-11-resistant cell lines. Adenovirus (AdGFPFasL(TET)) was used to deliver a Murine FasL-GFP fusion gene into human PCa cells resulting in 70-98% apoptosis at 48 h as determined by the MTS assay. DU145 and PPC-1 cells treated with AdGFPFasL(TET) stained positive for the TUNEL assay, indicating that cell death was via apoptosis. Using immunofluorescent microscopy, Fas and GFPFasL colocalized to the same intracellular compartment. The anti-Fas neutralizing antibody ZB-4 was unable to block AdGFPFasL(TET)-mediated cell death, suggesting that intracellular FasL may ligate Fas within the Golgi and/or endoplasmic reticulum. This is the first evidence suggesting that these two molecules interact prior to cell surface presentation. Collectively, these findings indicate that intracellular GFPFasL expression is superior to CH-11 at inducing Fas-mediated apoptosis in human PCa cells and may allow use of AdGFPFasL(TET) for PCa gene therapy. PMID- 11020351 TI - Targeting transgene expression to airway epithelia and submucosal glands, prominent sites of human CFTR expression. AB - Targeting therapeutic gene expression to disease-affected tissues is an essential component of effective and safe gene therapy. After birth, CFTR gene expression in human lungs is localized predominantly in the epithelial cells lining the upper airways, especially in the ducts and serous tubules of the submucosal glands. We have developed a K18 expression cassette, based on the DNA control elements of the human cytokeratin 18 gene. Temporal and spatial analyses of transgenic mice demonstrated that this expression cassette targets transgene expression to almost all cell types in which CFTR is expressed. Airway epithelium expression started as early as 11.5 days of gestational age and continued into the adulthood of the transgenic mice. In these adult mice, the pattern of the reporter expression strikingly matched that of the human cytokeratin 18 and human CFTR genes. The transgene expression was epithelium-specific and undetectable in connective tissue, muscle, bone, cartilage, blood, and endothelial cells. Significantly, high levels of expression were detected in tracheal submucosal glands. Together, these results suggest that our K18 expression cassette has a high potential for clinical application in gene therapy for patients with cystic fibrosis. PMID- 11020352 TI - Safety of adeno-associated virus as cochlear gene transfer vector: analysis of distant spread beyond injected cochleae. AB - The adeno-associated virus (AAV), inoculated into the perilymph, has been shown to be an effective vector for mediating intracochlear transgene expression. The unexpected finding of transgene expression in the contralateral cochlea in previous work raised concern about dissemination of the virus from the target tissue. The current study was undertaken to assess the extent of AAV dissemination following its introduction into the inner ear. Adult male guinea pigs were injected with recombinant AAV into their left ears and sacrificed at 2 or 4 weeks. Various organs including the cochleae were harvested to characterize the presence and expression of the viral DNA. Virus DNA was detected via polymerase chain reaction in the infused and contralateral cochlea and in the cerebellum but not in any other organs, including cortex, heart, lung, liver, spleen, and kidney. Although the viral presence was established in the cerebellum, transgene expression in this organ was undetectable with either Western blot or immunohistochemistry. Transgene expression was demonstrated via immunohistochemistry in multinucleated giant cells in the bone marrow spaces adjacent to the infused and contralateral cochleae. Collectively, these results suggest potential routes for AAV dissemination from the infused cochlea via the cochlear aqueduct or by extension through the temporal bone marrow spaces. This study reinforces the need to investigate factors that mitigate viral leakage. PMID- 11020353 TI - Reexpression following readministration of an adenoviral vector in adult mice after initial in utero adenoviral administration. AB - Adenovirus-mediated gene delivery is limited by the induction of immune responses that produce toxicity and prevent reexpression. To determine whether adenoviral delivery in the preimmune fetus would produce tolerance, we assessed luciferase (luc) expression following sequential pre- and postnatal adenoviral-mediated gene delivery. Day 15 fetuses were injected intrahepatically with 1 x 10(7) pfu of an adenoviral-luc vector (Ad-luc). Following in utero injection, hepatic luc expression persisted 1 month postnatally. No humoral response to adenovirus or luc was detected. Adult mice, previously injected in utero, were reinjected intravenously with 5 x 10(8) pfu of Ad-luc at 3 months of age and again at 6 months with either 5 x 10(8) pfu of Ad-luc or cationic liposome-DNA complexes (CLDC). Following the first postnatal injection, animals injected in utero had levels of luc comparable to those of age-matched naive controls. However, both control and experimental animals subsequently developed antibodies to adenovirus and luc. No further expression was achieved with a second postnatal injection of Ad-luc or with delivery of CLDC-luc. These studies demonstrate that the delivery of adenoviral vectors in utero at E15 does not elicit an immune response. However, delivery of recombinant adenovirus postnatally results in brisk and limiting immune responses regardless of the in utero exposure. PMID- 11020354 TI - Downregulation of CXCR4 gene expression in primary human endothelial cells following infection with E1(-)E4(+) adenovirus gene transfer vectors. AB - Infection of human endothelial cells with first-generation E1(-)E4(+) adenovirus (Ad) vectors leads to prolonged cell survival and changes in the cell phenotype to a more quiescent stage. Based on the concept that the CXCR4, the receptor for the endothelial chemoattractant stromal-derived factor-&alpha (SDF-alpha), is constitutively expressed by quiescent, resting endothelial cells, the present study analyzes the effect of Ad vector infection on CXCR4 expression and SDF alpha responses of human umbilical vein endothelial cells (HUVEC). CXCR4 transcripts were markedly downregulated in E1(-)E4(+) Ad-infected cells 48 h following infection, but not in uninfected control cells or when the cells were infected with an E1(-)E4(-) Ad vector. Analysis of surface CXCR4 expression by flow cytometry demonstrated marked reduction of the CXCR4 receptor on cells infected with E1(-)E4(+) Ad compared to uninfected control cells or E1(-)E4(-) Ad infected cells. Infection of other cell types which express CXCR4, such as dendritic cells and myeloma cells, did not exhibit CXCR4 receptor downregulation following infection with E1(-)E4(+) Ad. Consistent with the observed downregulation of CXCR4 mRNA and surface protein, infection of the endothelial cells with an E1(-)E4(+) Ad rendered the cells unresponsive to the chemoattractant SDF-alpha compared to naive or E1(-)E4(-) Ad-infected cells. Together, the data suggest that first-generation Ad vectors, likely the E4 region, modify the ability of endothelial cells to respond to at least one important chemoattractant. PMID- 11020355 TI - Effect of preexisting anti-herpes immunity on the efficacy of herpes simplex viral therapy in a murine intraperitoneal tumor model. AB - HSV-1716, a replicating nonneurovirulent herpes simplex virus type 1, has shown efficacy in treating multiple types of human tumors in immunodeficient mice. Since the majority of the human population has been previously exposed to herpes simplex virus, the efficacy of HSV-based oncolytic therapy was investigated in an immunocompetent animal tumor model. EJ-6-2-Bam-6a, a tumor cell line derived from h-ras-transformed murine fibroblast, exhibit a diffuse growth pattern in the peritoneal cavity of BALB/c mice and replicate HSV-1716 to titers observed in human tumors. An established intraperitoneal (ip) tumor model of EJ-6-2-Bam-6a in naive and HSV-immunized mice was used to evaluate the efficacy of single or multiple ip administrations of HSV-1716 (4 x 10(6) pfu/treatment) or of carrier cells, which are irradiated, ex vivo virally infected EJ-6-2-Bam-6a cells that can amplify the viral load in situ. All treated groups significantly prolonged survival versus media control with an approximately 40% long-term survival rate (cure) in the multiply treated, HSV-naive animals. Prior immunization of the mice with HSV did not significantly decrease the median survival of the single or multiply treated HSV-1716 or the carrier cell-treated groups. These studies support the development of replication-selective herpes virus mutants for use in localized intraperitoneal malignancies. PMID- 11020356 TI - Selective Rep-Cap gene amplification as a mechanism for high-titer recombinant AAV production from stable cell lines. AB - Gene transfer vectors based on adeno-associated virus mediate high-level, stable gene expression in a variety of postmitotic tissues; thus, there is interest in developing improved production systems. We previously described the generation of rAAV producer cell lines that, upon infection with adenovirus, yielded biologically active rAAV particles. In these studies we show that the adenovirus multiplicity of infection (m.o.i.) is a critical variable for efficient production of cell line-derived rAAV and can affect yields by over 200-fold. Moreover, a threshold level of adenovirus was found necessary for high-titer vector production. To define the possible factors responsible for adenovirus m.o.i. -dependent rAAV yields, we analyzed rep and cap expression as a function of adenovirus m.o.i. High-level AAV capsid protein synthesis was observed in rAAV producer cells at adenovirus m.o.i. > or =10. This prompted us to analyze the rep cap copy number following adenovirus infection. We documented robust episomal DNA amplification (100-fold) of integrated rep-cap sequences. Interestingly, no amplification of rep-cap sequences was observed when the sequences (in plasmid form) were transfected into adenovirus-infected HeLa cells. These data suggest that adenovirus-dependent rep-cap gene amplification is a critical process responsible for efficient rAAV synthesis in stable cell lines. PMID- 11020357 TI - Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes. AB - We exploited the ability of lentiviral vectors to govern the stable transduction of cells irrespective of their cycling status to induce the reversible immortalization of human primary cells. First, bicistronic HIV-derived lentiviral vectors expressing GFP- and the HSV1 thymidine kinase and containing the LoxP sequence in their LTR (HLox) were used to transduce HeLa cells. Cre expression led to efficient proviral deletion, and unexcised cells could be eliminated by ganciclovir treatment. A human liver biopsy was then exposed to a combination of HLox vectors that harbored either the SV40 large T (TAg) or the human telomerase (hTERT) DNAs in place of GFP. This led to the isolation of liver sinusoidal endothelial cell (LSEC) clones that exhibited an immortalized phenotype while retaining most of the features of primary hLSEC. Complete growth arrest of these cells was observed in 2 days of Cre expression, and the resulting stationary culture could be kept for at least 2 weeks. Transduction of human adult pancreatic islets with HLox vectors coding for Tag and Bmi-1 also induced the proliferation of insulin-positive cells. These results indicate that lentivectors can be used to mediate the reversible immortalization of primary nondividing cells and should allow for the production of large supplies of a wide variety of human cells for both therapeutic and research purposes. PMID- 11020359 TI - An enzyme-linked immunosorbent assay for the detection of antibody to avian reovirus by using protein sigma B as the coating antigen. AB - An enzyme-linked immunosorbent assay using the expressed protein sigma B as the coating antigen (sigma B-ELISA) for detecting antibody to avian reovirus (ARV) in chickens was developed and compared with a conventional ELISA. Both ELISA s and a serum neutralisation (SN) test were used to test the sera from experimentally vaccinated and farm chickens. The sigma B-ELISA could clearly distinguish the SN positive and -negative sera in 38-week-old chickens. The correlation rate between SN and a sigma B-ELISA was 100 per cent (65/65), and that between SN and conventional ELISA was 84 per cent (55/65). With the sigma B-ELISA, all SN negative sera had low absorbance values (below 0.06), and the absorbance values correlated closely with the SN titres. However, the sera which were antibody negative by SN had various absorbance values, ranging from 0.07 to 0.39 in the conventional ELISA. Hence, the sigma B-ELISA had lower non-specific binding reactions than the conventional ELISA against sera from ARV -negative birds. Antibody against ARV could be detected by sigma B-ELISA after vaccination. Absorbance values peaked 4 weeks after vaccination at 2 weeks of age and were maintained until the birds were 27 weeks old. The results suggest that the presence of antibody against viral protein sigma B in birds may be used as a good indicator by the sigma B - ELISA for detecting immune status of a chicken flock or to detect chickens infected with ARV. PMID- 11020358 TI - Only one medicine: the future of comparative medicine and clinical research. AB - This article reviews the roots of comparative medicine and argues that during the 20th century it failed to realise its full potential. New opportunities arise from the growing availability of precise, minimally invasive, clinically compatible techniques, which enable us to benefit from the availability of spontaneous analogues of human disease in animals. Particularly with multifactorial diseases, these offer a unique blend of authenticity and acceptability. To realise the full benefits to both animals and humans, we need much closer alignment of human and veterinary clinical medicine. PMID- 11020361 TI - Ameliorative effects of sodium bentonite on phagocytosis and Newcastle disease antibody formation in broiler chickens during aflatoxicosis. AB - The ameliorative effect of graded levels of dietary sodium bentonite (0.2, 0.4 and 0.6 per cent wt/wt of feed) on in vitro-impaired phagocytosis and suppressed immune response to Newcastle disease vaccine during aflatoxicosis (AF) in broiler chicks was investigated. Both percentage and mean of phagocytic activities were decreased significantly (P < 0.05) in chicks fed 2.5 mg aflatoxin per kg feed. The addition of sodium bentonite was significantly effective in ameliorating the negative effect of AF on the percentage and mean of phagocytosis. The presence of AF alone in the diet depressed the immune response of chicks as measured by haemagglutination inhibition (HI) test. Sodium bentonite was also effective in ameliorating the suppressive effect of AF on the HI -titre in chicks vaccinated against Newcastle disease. The best results obtained when sodium bentonite was added at the rate of 0.4 per cent wt/wt of feed to the AF-containing diets. PMID- 11020360 TI - Localization of the lectin reactive sites in adult and prepubertal horse testes. AB - The testes of prepubertal and adult horses were investigated using 10 horseradish peroxidase conjugated lectins combined with sialidase digestion and potassium hydroxide treatment, to localise the oligosaccharide sequences of glycoconjugates during spermatid maturation. In adult animals, the lectins showed a variable affinity for spermatids and Sertoli cell apical extensions. Soybean agglutinin (SBA), peanut agglutinin (PNA), Ricinus communis agglutinin (RCA-I) and wheat germ agglutinin (WGA) bound to the acrosomal structures of spermatids, whereas Griffonia simplicifolia agglutinin (GSA-II) labelled these structures only during Golgi and cap phases. These results suggested that glycoproteins of mature acrosomes contain both N- and O-linked oligosaccharides and that these carbohydrate chains undergo modifications during spermiogenesis. Sialic acid residues were not detected throughout the acrosomal development. The lectin binding pattern of Sertoli cells was very similar to that of acrosome of spermatids during the maturation phase. In sexually immature horses, only the degenerated germinal cells and the Leydig cells showed reactivity towards lectins. The first cells reacted with SBA and Dolichos biflorus agglutinin (DBA), the latter with SBA, PNA, WGA, GSA-II, Canavalia ensiformis agglutinin (ConA), Lens culinaris agglutinin (LCA) and also with DBA after sialidase digestion. PMID- 11020362 TI - Urinary markers of type I collagen degradation in the dog. AB - Urinary assays for type I collagen metabolites provide a non invasive index of bone resorption in humans, and are widely used in the management of patients with metabolic bone diseases. The specific aims of this study were to investigate the feasibility of using commercial human assay kits for quantifying the urinary excretion of type I collagen metabolites in dogs of different ages. Urine and serum samples were collected from 35 beagle dogs in five age groups (0 to 1 years; 1 to 2 years; 2 to 3 years; 3 to 7 years; > 8 years old). Urinary concentrations of pyridinoline (Pyd), deoxypyridinoline (Dpd), and the carboxy- and amino-terminal cross-linked telopeptides of type I collagen (CTx and NTx, respectively) were measured with commercial enzyme-linked immunoassay kits. Serum concentrations of another type I collagen metabolite, the carboxy-terminal cross linked teloptide of type I collagen (ICTP), were measured with a commercial radioimmunoassay. Dilutional studies indicated that the four urinary assays show specific cross-reactivity with canine urine. Age-related differences in urinary marker excretion were identified, with young dogs excreting the highest concentrations of Pyd, Dpd, NTx and CTx. The correlation between the individual urinary markers was excellent (r = 0.87 to 0.98), while the correlation between serum ICTP and individual urinary markers was weaker (r = 0.52 to 0.64). These results validate the usefulness of the commercial assay kits in monitoring type I collagen metabolism in dogs. Histomorphometric studies have confirmed the relationship between collagen degradation and bone resorption in humans, and similar studies are now needed in dogs. PMID- 11020364 TI - Responses of adult cattle to vaccination with a reduced dose of Brucella abortus strain RB51. AB - The efficacy of a reduced dosage of Brucella abortus strain RB51 (SRB51) was evaluated in adult cattle. Hereford heifers were vaccinated with saline, or three SRB51 dosing regimens (3 x 10(9) colony forming units (cfu) once, 1 x 10(9) cfu once, or 1 x 10(9) cfu twice). Cattle vaccinated with 3 x 10(9) cfu of SRB51 had greater (P < 0.05) antibody responses than non-vaccinates, and greater lymphocyte proliferative responses to SRB51 than all other treatments. Four of six non vaccinated heifers aborted after mid-gestational challenge with B abortus strain 2308 (S2308) and S2308 was recovered from tissues obtained from all the non vaccinates. In comparison, S2308 or SRB51 were not recovered from tissues from heifers vaccinated with any of the SRB51 dosage regimens. The data suggest that vaccination with a reduced dosage of SRB51 protects adult cattle against abortion or infection caused by exposure to virulent B abortus during the subsequent pregnancy. PMID- 11020363 TI - Effects of lorglumide and atropine on MgSO(4)-induced gallbladder emptying in conscious dogs. AB - The aim of the study was to examine the possible involvement of cholecystokinin (by lorglumide) and cholinergic mechanisms (by atropine) in magnesium sulphate (MgSO(4))-induced gallbladder contraction of conscious dogs. The gallbladder (GB) volume was determined by ultrasonography. The optimal dose of 80 mg kg(-1)of MgSO(4)was determined from a MgSO(4)dose-response curve using doses of 10, 20,40, 80, 120 mg kg(-1). The largest dose of MgSO(4)was less effective than the optimal dose. Peak gallbladder contraction (32 per cent) was achieved at 30 minutes. Atropine (50 microg kg(-1)s.c.) or lorglumide (1 mg kg(-1)p.o.) fully prevented GB contraction. In conclusion, supraoptimal doses of MgSO(4)have a diminishing effect. The sustained contraction of the gallbladder in response to the optimal dose of MgSO(4)can be explained by an additive effect of the cholecystokinin release and a cholinergic trigger mechanism. Ultrasonography and MgSO(4)stimulation proved to be a valuable technique for examination of gallbladder motility. PMID- 11020365 TI - Haematological profile of crossbred dairy cattle to monitor herd health status at medium elevation in Central Himalayas. AB - Haematological profile-haemoglobin concentration (Hb), total erythrocytes count (TEC), packed cell volume (PCV), erythrocyte indices-mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were studied in crossbred dairy cattle (Holstein Friesian x Sahiwal) under various physiological states: non-pregnant heifers (NPH), pregnant heifers (PH), empty dry cows (EDC), pregnant lactating cows (PLC), medium yield early lactating cows (MYELC) and high yield early lactating cows (HYELC) during summer and winter seasons at 1700 metres altitude from mean sea level in the Central Himalayas. On comparison of annual means, the highest values of Hb and PCV were recorded in PH and of TEC in NPH, whereas the lowest values of these parameters were found in EDC. The Hb and TEC tended to decrease with increasing milk yield. Comparison of annual means of erythrocyte indices revealed the highest MCV and MCH in EDC, which simultaneously showed the lowest MCHC. Significant seasonal variations in haematological profile were recorded. The overall group mean (OGM) of Hb, MCV, MCH and MCHC was found to be significantly higher (P < 0.01) during summer whereas the TEC and PCV showed higher OGM (P < 0.01) during the winter season. PMID- 11020366 TI - The post-operative analgesic effects of ketamine after canine ovariohysterectomy- a comparison between pre- or post-operative administration. AB - Thirty-six female dogs undergoing ovariohysterectomy were randomly allocated into three groups in this assessor-blinded study. The control group received no ketamine, the preketamine group were given ketamine (2.5 mg kg(-1)i.m.) at anaesthetic induction (in addition to the induction agents), the post-ketamine group received ketamine (2.5 mg kg(-1)i.m.) at extubation. Mechanical nociceptive thresholds and visual analogue scale (VAS) scores were measured before premedication and post-operatively at 20 minutes, 1, 2, 4, 8 and 18 hours after extubation. Dogs in the control group required more rescue analgesics than those in the other two groups (significantly more than the preketamine group), they also had consistently higher VAS pain scores throughout the post-operative period. Administration of ketamine post-operatively delayed the onset of post operative wound hyperalgesia; dogs in the control group had the greatest amount of post-operative wound hyperalgesia. A single subanaesthetic dose of ketamine provided effective but short acting analgesia and preoperative administration may confer some benefits over administration post-operatively. PMID- 11020367 TI - Serum haptoglobin: an objective indicator of experimentally-induced Salmonella infection in calves. AB - Experimental models of Salmonella -induced gastroenteritis have previously relied on crude subjective clinical markers of infection to assess disease severity. The aim of this study was to investigate the possibility that changes in serum levels of the acute phase protein, haptoglobin, may be used as an objective, quantitative measurement of infection. Eight 3- to 4-week-old animals were challenged with a mixture of three Salmonella serotypes containing 6 x 10(10)bacteria and compared with five animals given a placebo preparation. Animals were monitored and characteristic clinical symptoms of infection; diarrhoeal scores, morbidity scores and rectal temperature, were recorded. Serum samples, from both animal groups, taken prior to challenge and again on days 1, 3, and 5 post-challenge, were analysed for haptoglobin levels using a direct serum binding assay. Prior to challenge, all 13 animals had normal levels of haptoglobin in their serum. By day 3 post-challenge six of eight animals challenged with Salmonella had abnormal serum haptoglobin levels (median level = 212 microg ml(-1)), while haptoglobin levels remained normal in placebo challenged animals (median level = 0 microg ml(-1)). The change in haptoglobin levels during the 5-day observation period was statistically significant in the Salmonella -challenged animals (P = 0.0003, H = 16.477). Serum haptoglobin levels showed a statistical correlation with clinical measures of disease severity; diarrhoeal scores (P = 0.0015, H =8. 988), morbidity scores (P = 0.0004, H = 15.711) and rectal temperature (P = 0.0001, Z = 4.304). Thus, serum haptoglobin levels closely reflect the clinical symptoms of infection and are therefore a useful marker of infection severity in salmonellosis in calves. PMID- 11020368 TI - Characterisation of sugar residues in glycoconjugates of pig mandibular gland by traditional and lectin histochemistry. AB - Sugar residues are important components of salivary gland secretion. Traditional histochemical methods and lectin histochemistry were used to characterise glycoconjugates present in the mandibular gland of normal adult pigs. Acinar cells contained abundant quantities of glycoconjugates with the terminal trisaccharide sialic acid - (alpha 2-->3, 6) galactosyl (beta 1-->3) N acetylgalactosamine. Mandibular acinar cells also contained alpha and beta N acetylgalactosamine and N -acetylglucosamine residues, whereas the demilunar cells contained glycoconjugates with fucose, mannose and N -acetylglucosamine residues. In the duct system a range of sugar residues were localised throughout the cell cytoplasm or limited to the apical surface. These results provide new knowledge concerning the structure of salivary glycoconjugates in normal adult pig and a basis for future pathological studies. PMID- 11020369 TI - Difficulties in experimental infection studies with Flavobacterium psychrophilum in rainbow trout (Oncorhynchus mykiss) using immersion, oral and anal challenges. AB - This study was carried out in order to try to establish an efficacious and reliable experimental infection model for Flavobacterium psychrophilum, the causative agent of rainbow trout fry syndrome, using contact, oral and anal challenges. Ten F psychrophilum strains of different origin were included. The influence of water temperature, scarification, water quality, stress and growth conditions of the pathogen on the experimental infection was assessed. For each challenge protocol, all strains failed to reproduce disease signs or mortality in rainbow trout (Oncorhynchus mykiss L.) fry. Histological and bacteriological examination of the skin, gills and internal organs of the fish 3 weeks following inoculation were found to be negative. Different hypotheses to explain the inability of the challenge models to reproduce the disease experimentally are discussed. PMID- 11020370 TI - Genomic polymorphism in Uruguayan Creole cattle using RAPD and microsatellite markers. AB - Uruguayan Creole cattle inhabit areas that cannot sustain conventional farming. They have adapted to fragile environments and are influenced only by natural selection. In this study, random amplified polymorphic DNA (RAPD) and microsatellite (MS) markers were used to analyse Creole cattle genome polymorphism. A comparative analysis using the RAPD technique was performed in pooled DNA of three cattle breeds (Holstein Friesian, Creole and Hereford) in order to evaluate their amplification patterns. A primary screening of RAPD primers allowed us to select and use those with higher percentage of GC base composition. A total of 215 loci ranging between 300 and 2500 bp were amplified. Bandsharing frequency (BSF) among breeds showed that less related fingerprints were observed between Creole and Hereford cattle (0.77), while the highest similarity frequency corresponded to Holstein Friesian compared to Hereford (0.81). Specific RAPD bands were identified in the three DNA pools and they were tested in every individual of each breed. It may be possible to isolate and sequence these bands to create breed-specific molecular markers. The identification of multiple alleles of the MSCYP 21 in Creole cattle with an heterozygosity of He = 0.846 supported the variability of this genetic resource. The use of molecular markers such as RAPD s and microsatellites is proposed to establish genetic distance among American Creole cattle and possibly related ancestral Iberian breeds. PMID- 11020371 TI - Rapid changes occur in the percentage of circulating bovine WC1(+)gamma delta Th1 cells. AB - gamma delta T cells found in the peripheral blood of cattle include a major subpopulation distinguished by expression of WC1. These cells are distinct from the WC1(-)gamma delta T cell population based on T cell receptor gene usage. We documented that a group of 6-month-old calves allowed free-range grazing and access to their mothers had a significantly greater proportion of total gamma delta T cells in their blood, attributable to the WC1(+)gamma delta T cell subpopulation, compared to age and breed-matched calves held in conventional housing. When the animals with the greater proportion of gamma delta T cells were transferred to conventional housing there was a decrease in the WC1(+)population so that by 3 weeks after transfer there was no longer a significant difference between the two groups. To investigate the biological activities of WC1(+)gamma delta T cells, the cells were purified by flow cytometric sorting. In vitro, they responded to stimulation by irradiated monocytes in autologous mixed leukocyte reaction (AMLR) cultures but not to direct stimulation through the T cell receptor (T c R) by anti-delta monoclonal antibody. After stimulation in the AMLR, WC1(+)gamma delta T cells had a Th1 cytokine profile characterised by production of IFN -gamma and lack of IL -4. Thus we propose that higher levels of the WC1(+)gamma delta T cells may provide calves with a mechanism to produce Th1 cytokines and that the level of these cells may be modulated according to environment or stress since both groups of calves were apparently disease-free. PMID- 11020372 TI - Systemic and local immune response of cows to intramammary infection with Staphylococcus aureus. AB - The association between Staphylococcus aureus chronic mammary gland infection and the resulting immune response expressed by the production of specific IgG and IgA antibodies in blood and milk was studied in Israeli Holstein cows. Specific antibodies of the IgG class were detected in sera of 82.6 per cent of the cows chronically infected by S aureus, while in 17.4 per cent no such antibodies could be detected. Specific IgG antibodies to S aureus were neither detected in sera of cows free of mammary infection nor in those infected with different coagulase negative staphylococci (CNS) such as S intermedius, S chromogenes or S haemolyticus. In milk, specific IgG antibodies to S aureus were detected only in cows with positive serology. The end point dilutions in the milk were 5 to 30 per cent of that of blood from the same cow. No significant difference in IgG titres was found in the same cow if the quarter was infected with S aureus or not. Specific antibodies to S aureus of the IgA class could not be detected in the sera of any of the cows included in this study. In milk, a specific IgA antibody was detected only in the samples from the S aureus infected quarters in which S aureus was isolated at the time of the experiment. In the same cow, quarters infected by S aureus were found to have a significantly higher IgA titre (P < 0.0001) than that of the non-infected ones. PMID- 11020373 TI - Detection of IL-2 and IFN-gamma m RNA expression in bovine milk cells at the late stage of the lactation period with RT-PCR. AB - The expression of bovine interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) in the late stage of the lactation period in milk cells was detected with reverse transcriptase-polymerase chain reaction (RT-PCR). IFN-gamma was detected with all primer concentrations, 0.2, 0.5 and 1 microM. IL-2 however, was faintly detected only with 0.5 microM primer concentration. It was shown that IFN-gamma m RNA expression can be demonstrated clearly with RT-PCR at the late stage of the lactation period. In contrast, IL-2 was weakly expressed at this stage of the lactation period. The work is important for substantiating the therapeutic use of recombinant bovine IFN-gamma and IL-2 and for helping to reveal the activity of cytokine in the mammary gland at the late stage of the lactation period. PMID- 11020374 TI - The importance of immediate destruction in epidemics of foot and mouth disease. AB - Foot and mouth disease is still prevalent in many parts of the world, as emphasised by the recent devastating epidemic in pig farms in Taiwan. A discrete time mechanistic model has been used to describe the spread of infection in both this epidemic and the 1967 to 1968 epidemic in the UK. The force of infection and basic reproduction number are estimated and the sensitivity of these results to the distributions of both the latent and infectious periods of the disease is examined. Epidemic simulations were performed to evaluate the disease control policy whereby all herds are slaughtered on the same day as disease confirmation. These simulations showed that implementing this policy could have resulted in a dramatic reduction (of over 60 per cent) in the number of pig farms affected in the Taiwan epidemic. It is thus imperative that the necessary resources are available to implement this policy, should an outbreak occur. PMID- 11020375 TI - Preventive efficacy of clinoptilolite in broilers during chronic aflatoxin (50 and 100 ppb) exposure. AB - This study was conducted to evaluate the protective efficacy of clinoptilolite (CLI, a natural zeolite) in the prevention of toxic effects of aflatoxin (AF). A total of 576 1-day-old Ross broiler chicks were housed in six treatment groups [six replicates of 16 each; control, CLI (15 g kg(-1)diet), 50 parts per billion (ppb) AF, 50 ppb AF plus CLI, 100 ppb AF, 100 ppb AF plus CLI ] for 42 days. Compared to controls, 100 ppb AF treatment significantly decreased body weight gains of chicks. The addition of CLI to the 100 ppb AF - containing diet moderately reduced the adverse effects of AF on performances of chicks. The chicks consuming 50 ppb AF -containing diet showed no significant differences on investigated parameters compared to controls. Also, the single addition of CLI to the AF -free diet had no adverse effects in birds. These results suggest that CLI can be beneficial in broilers for protection of AF toxicity at the level above. PMID- 11020376 TI - Role of Ca2+in the replication and pathogenesis of rotavirus and other viral infections. AB - Ca2+ plays a key role in many pathological processes, including viral infections. Rotavirus, the major etiological agent of viral gastroenteritis in children and young animals, provides a useful model to study a number of Ca2+ dependent virus cell interactions. Rotavirus entry, activation of transcription, morphogenesis, cell lysis, particle release, and the distant action of viral proteins are Ca2+ dependent processes. In the extracellular medium, Ca2+ stabilizes the structure of the viral capsid. During entry into the cell the low cytoplasmic Ca2+ concentration induced the solubilization of the outer protein layer of the capsid and transcriptase activation. Viral protein synthesis modifies Ca2+ homeostasis which, in turn, favours viral morphogenesis and induces cell death. The generation of diarrhea is a multifactorial process involving Ca2+ dependent secretory processes of mediators and water and electrolytes, as well as the induction of cell death in the different cell types that compose the intestinal epithelium. The discovery of the non-structural viral protein NSP4 as a viral enterotoxin and the possible participation of the enteric nervous system in the pathogenesis of diarrhea represent significant advances in its understanding. Ca2+ also plays a role in the replication cycles and pathogenesis of other viral diseases such as poliovirus, Coxsackie virus, cytomegalovirus, vaccinia and measles virus and HIV. PMID- 11020377 TI - Positive regulation of capacitative Ca2+ entry by intracellular Ca2+ in Xenopus oocytes expressing rat TRP4. AB - We have investigated the role of intracellular Ca2+ in the opening of capacitative Ca2+ entry (CCE) channels formed with rat TRP4 (rTRP4) using Xenopus oocytes. In rTRP4-expressing oocytes pretreated with thapsigargin, perfusion with A23187, a Ca2+ ionophore, significantly potentiated the delayed phase of the CCE mediated Cl- current response evoked by extracellular perfusion with Ca2+, without affecting the transient phase of CCE response. In control oocytes, the potentiation of delayed CCE response by A23187 was not significant. Using cut open recording in combination with artificial intracellular perfusion of oocytes, CCE-mediated Cl- response was recorded at controlled cytosolic Ca2+ concentrations. Intracellular perfusion with a Ca2+ free solution containing 10 mM EGTA abolished most of the CCE responses of both non-injected and rTRP4 expressing oocytes. The native CCE response was not fully recovered by subsequent increases in the intracellular Ca2+ concentration up to 300 nM. However, CCE response of the rTRP4-expressing oocytes was restored at an internal Ca2+ concentration of 110 nM. Blockade of endogenous Cl- channels with anion channel blocker isolated Ca2+ current flowing through CCE channels and clarified the difference in the sensitivity to an internal Ca2+ concentration. These findings indicate that recombinant CCE channels formed with rTRP4 are positively regulated by cytosolic Ca2+ at higher sensitivity compared to oocyte-endogenous CCE channels. PMID- 11020378 TI - Evidence for a vesicle-mediated maintenance of store-operated calcium channels in a human embryonic kidney cell line. AB - Direct microinjection of the clostridial neurotoxins botulinum neurotoxin A light chain or tetanus neurotoxin into cells of a human embryonic kidney cell line significantly reduced calcium entry after depletion of internal calcium stores by cyclopiazonic acid, a reversible inhibitor of the sarcoplasmic-endoplasmic reticular calcium-ATPases. Botulinum neurotoxin A light chain specifically hydrolyzes a synaptosomal-associated protein of 25 kilodaltons (SNAP-25), and tetanus neurotoxin specifically hydrolyzes synaptobrevin-2 (vesicle-associated membrane protein 2, VAMP-2) and cellubrevin (vesicle-associated membrane protein 3, VAMP-3). Since these substrate proteins are required for vesicle docking and fusion, inhibition of store-operated calcium entry by botulinum neurotoxin A light chain and tetanus neurotoxin supports a model in which vesicle fusion is a prerequisite for activation of store-operated calcium entry. Brefeldin A, a fungal metabolite that interferes with vesicle traffic, partially reduced calcium entry following store depletion. The size of the reserve pool of vesicles or parallel vesicle recycling pathways employing brefeldin A-sensitive and brefeldin A-insensitive ADP-ribosylation factors may explain the failure of brefeldin A to completely inhibit store-operated calcium entry. PMID- 11020379 TI - Spontaneous calcium oscillatory patterns in mammotropes display non-random dynamics. AB - We previously showed that primary rat mammotropes exhibited four distinct patterns of 'spontaneous' free intracellular calcium ([Ca2+]i) oscillatory behavior: a quiescent state A and three oscillatory states B,C&D, which differed in frequency/amplitude characteristics. When [Ca2+]i was monitored in 10 min windows separated by several hours, these phenotypes were frequently found to interconvert, raising the question about whether these transitions were random or ordered events. We reasoned that if such activity were random, then neither episode duration nor transitional probabilities should differ among phenotypes. We tested this logic in the current study by making long-term, continuous measurements of [Ca2+]i in mammotropes microinjected with Fura-2-dextran and identified by their ability to express a prolactin promoter-driven reporter plasmid. We found that transitions occurred in ~25% of cells (n = 36 from 9 independent experiments) once every 1-5 h and demarcated phenotype episodes of different duration (A, 1.04 +/- 0.2 h; B, 1.64 +/- 0.3 h; C, 2.45 +/- 0.62 h; D, 0.90 +/- 0.2 h, mean +/- SEM). Moreover, some transitions occurred more frequently than others and linked specific phenotypes into a common pattern: C to B to A. Our results demonstrate that the seemingly spontaneous nature of [Ca2+]i phenotype transitions are, in fact, ordered and support the view that they comprise a structured 'code' like that proposed to underlie calcium-dependent regulation of exocytosis and gene expression. PMID- 11020380 TI - Induction of spontaneous recurrent epileptiform discharges causes long-term changes in intracellular calcium homeostatic mechanisms. AB - Calcium and calcium-dependent systems have been long implicated in the induction of epilepsy. We have previously observed that intracellular calcium ([Ca2+]i) levels remain elevated in cells undergoing epileptogenesis in the hippocampal neuronal culture (HNC) model. In this study, we employed the hippocampal neuronal culture (HNC) model of in vitro 'epilepsy' which produces spontaneous recurrent epileptiform discharges (SREDs) for the life of the neurons in culture to investigate alterations in [Ca2+]i homeostatic mechanisms that may be associated with the 'epileptic' phenotype. [Ca2+]i imaging fluorescence microscopy was performed on control and 'epileptic' neurons with two different fluorescent dyes ranging from high to low affinities for [Ca2+]i. We measured baseline [Ca2+]i levels and the ability to restore resting [Ca2+]i levels after a brief 2-min exposure to the excitatory amino acid glutamate in control neurons and neurons with SREDs. Neurons manifesting SREDs had statistically significantly higher baseline [Ca2+]i levels that persisted for the life of the culture. In addition, the 'epileptic' phenotype was associated with an inability to rapidly restore [Ca2+]i levels to baseline following a glutamate induced [Ca2+]i load. The use of the low affinity dye Fura-FF demonstrated that the difference in restoring baseline [Ca2+]i levels was not due to saturation of the high affinity dye Indo 1, which was utilized for evaluating the [Ca2+]i kinetics at lower [Ca2+]i levels. Peak [Ca2+]i levels in response to glutamate were the same in both 'epileptic' and control neurons. While [Ca2+]i levels recovered in approximately 30 min in control cells, it took more than 90 min to reach baseline levels in cells manifesting SREDs. Alterations of [Ca2+]i homeostatic mechanisms observed with the 'epileptic' phenotype were shown to be independent of the presence of continuous SREDs and persisted for the life of the neurons in culture. Epileptogenesis was shown not to affect the degree or duration of glutamate induced neuronal depolarization in comparing control and 'epileptic' neurons. The results indicate that epileptogenesis in this in vitro model produced long lasting alterations in [Ca2+]i regulation that may underlie the 'epileptic' phenotype and contribute to the persistent neuroplasticity changes associated with epilepsy. PMID- 11020381 TI - Cholinergic control of membrane conductance and intracellular free Ca2+ in outer hair cells of the guinea pig cochlea. AB - We have studied the action of cholinergic agonists on outer hair cells, both in situ and isolated from the cochlea of the guinea pig, combining new fast CCD technology for Ca2+ imaging and conventional patch-clamp methods. Carbachol (1 mM) activated a current with a reversal potential near -70 mV and a bell-shaped I V curve, suggesting that it was a Ca2+ activated K+ current. In a few cells, this current was preceded by a transient inward current, probably owing to an influx of Ca2+ and other cations through the acetylcholine (ACh) receptors. The amplitude of the Ca2+ signal was maximal in a circumscribed region at the basal pole of the cell and decreased steeply towards the apical pole, compatible with Ca2+ influx and/or Ca2+ induced Ca2+ release at the cells base. The time course of the Ca2+ rise was fastest at the base, but it was still slightly slower, and more rounded, than that of the K+ current. In some recordings the K+ current was observed without any measurable change of intracellular Ca2+. The K+ current was potentiated (18%) by caffeine (5 mM), and decreased (19%) by ryanodine (0.1 mM) in the majority of cells tested. The results are discussed in terms of a labile intracellular Ca2+ store located at the base of the cell, close to the Ca2+ permeable ACh receptor channels and Ca2+ activated K+ channels, whose contribution to the Ca2+ rise occurring in the region of the channels is variable, and probably dependent on its ability to refill with Ca2+. PMID- 11020382 TI - Lipoarabinomannan from Mycobacterium tuberculosis promotes macrophage survival by phosphorylating Bad through a phosphatidylinositol 3-kinase/Akt pathway. AB - Efforts in prevention and control of tuberculosis suffer from the lack of detailed knowledge of the mechanisms used by pathogenic mycobacteria for survival within host cell macrophages. The exploitation of host cell signaling pathways to the benefit of the pathogen is a phenomenon that deserves to be looked into in detail. We have tested the hypothesis that lipoarabinomannan (LAM) from the virulent species of Mycobacterium tuberculosis possesses the ability to modulate signaling pathways linked to cell survival. The Bcl-2 family member Bad is a proapoptotic protein. Phosphorylation of Bad promotes cell survival in many cell types. We demonstrate that man-LAM stimulates Bad phosphorylation in a phosphatidylinositol 3-kinase (PI-3K)-dependent pathway in THP-1 cells. Man-LAM activated PI-3K. LAM-stimulated phosphorylation of Bad was abrogated in cells transfected with a dominant-negative mutant of PI-3K (Delta p85), indicating that activation of PI-3K is sufficient to trigger phosphorylation of Bad by LAM. Since phosphorylation of Bad occurred at serine 136, the target of the serine/threonine kinase Akt, the effect of LAM on Akt kinase activity was tested. Man-LAM could activate Akt as evidenced from phosphorylation of Akt at Thr(308) and by the phosphorylation of the exogenous substrate histone 2B. Akt activation was abrogated in cells transfected with Deltap85. The phosphorylation of Bad by man LAM was abrogated in cells transfected with a kinase-dead mutant of Akt. These results establish that LAM-mediated Bad phosphorylation occurs in a PI-3K/Akt dependent manner. It is therefore the first demonstration of the ability of a mycobacterial virulence factor to up-regulate a signaling pathway involved in cell survival. This is likely to be one of a number of virulence-associated mechanisms by which bacilli control host cell apoptosis. PMID- 11020383 TI - 2-acetylaminofluorene up-regulates rat mdr1b expression through generating reactive oxygen species that activate NF-kappa B pathway. AB - Overexpression of multidrug resistance genes and their encoded P-glycoproteins is a major mechanism for the development of multidrug resistance in cancer cells. The hepatocarcinogen 2-acetylaminofluorene (2-AAF) efficiently activates rat mdr1b expression. However, the underlying mechanisms are largely unknown. In this study, we demonstrated that a NF-kappa B site on the mdr1b promoter was required for this induction. Overexpression of antisense p65 and I kappa B alpha partially abolished the induction. We then delineated the pathway through which 2-AAF activates NF-kappa B. 2-AAF treatment led to the increase of intracellular reactive oxygen species (ROS) which causes activation of IKK kinases, degradation of I kappa B beta (but not I kappa B alpha), and increase in NF-kappa B DNA binding activity. Consistent with the idea that ROS may participate in mdr1b regulation, antioxidant N-acetylcysteine inhibited the induction of mdr1b by 2 AAF. Overproduction of a physiological antioxidant glutathione (GSH) blocked the activation of IKK kinase complex and NF-kappa B DNA binding. Based on these results, we conclude that 2-AAF up-regulates mdr1b through the generation of ROS, activation of IKK kinase, degradation of I kappa B beta, and subsequent activation of NF-kappa B. This is the first report that reveals the specific cis elements and signaling pathway responsible for the induction of mdr1b by the chemical carcinogen 2-AAF. PMID- 11020384 TI - A transmembrane site determines sensitivity of neuronal nicotinic acetylcholine receptors to general anesthetics. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are potential targets for a wide variety of general anesthetics. We recently showed that alpha(4)beta(2) nAChRs are more sensitive than alpha(4)beta(4) receptors to the gaseous anesthetics nitrous oxide and xenon. The present study examines chimeric and point mutant rat nAChRs expressed in Xenopus oocytes and identifies a single amino acid residue (beta(2)-Val(253) or beta(4)-Phe(255)) near the middle of the second transmembrane segment (TM2) that determines gaseous anesthetic sensitivity. Mutations of this residue in beta subunits and the homologous residue of alpha(4) subunits (alpha(4)-Val(254)) showed that this position also determines sensitivities of nAChRs to acetylcholine, isoflurane, pentobarbital, and hexanol. In contrast, these mutations did not affect actions of ketamine. The positively charged sulfhydryl-specific reagent methanethiosulfonate ethylammonium reacted with a cysteine introduced at alpha(4)-Val(254) or beta(2)-Val(253), and irreversibly reduced anesthetic sensitivities of nAChRs. Propyl methanethiosulfonate is an anesthetic analog that covalently binds to a TM2 site of gamma-aminobutyric acid(A) and glycine receptors and irreversibly enhances receptor function. However, propyl methanethiosulfonate reversibly inhibited cysteine-substitution mutants at alpha(4)-Val(254) or beta(2)-Val(253) of nAChRs, and did not affect anesthetic sensitivity. Thus, residues alpha(4)-Val(254) and beta(2)-Val(253) alter channel gating and determine anesthetic sensitivity of nAChRs, but are not likely to be anesthetic-binding sites. PMID- 11020385 TI - Two-step processing of human frataxin by mitochondrial processing peptidase. Precursor and intermediate forms are cleaved at different rates. AB - We showed previously that maturation of the human frataxin precursor (p-fxn) involves two cleavages by the mitochondrial processing peptidase (MPP). This observation was not confirmed by another group, however, who reported only one cleavage. Here, we demonstrate conclusively that MPP cleaves p-fxn in two sequential steps, yielding a 18,826-Da intermediate (i-fxn) and a 17,255-Da mature (m-fxn) form, the latter corresponding to endogenous frataxin in human tissues. The two cleavages occur between residues 41-42 and 55-56, and both match the MPP consensus sequence RX downward arrow (X/S). Recombinant rat and yeast MPP catalyze the p --> i step 4 and 40 times faster, respectively, than the i --> m step. In isolated rat mitochondria, p-fxn undergoes a sequence of cleavages, p - > i --> m --> d(1) --> d(2), with d(1) and d(2) representing two C-terminal fragments of m-fxn produced by an unknown protease. The i --> m step is limiting, and the overall rate of p --> i --> m does not exceed the rate of m --> d(1) --> d(2), such that the levels of m-fxn do not change during incubations as long as 3 h. Inhibition of the i --> m step by a disease-causing frataxin mutation (W173G) leads to nonspecific degradation of i-fxn. Thus, the second of the two processing steps catalyzed by MPP limits the levels of mature frataxin within mitochondria. PMID- 11020386 TI - Excluded volume effects on the refolding and assembly of an oligomeric protein. GroEL, a case study. AB - We have studied the effect of macromolecular crowding reagents, such as polysaccharides and bovine serum albumin, on the refolding of tetradecameric GroEL from urea-denatured protein monomers. The results show that productive refolding and assembly strongly depends on the presence of nucleotides (ATP or ADP) and background macromolecules. Nucleotides are required to generate an assembly-competent monomeric conformation, suggesting that proper folding of the equatorial domain of the protein subunits into a native-like structure is essential for productive assembly. Crowding modulates GroEL oligomerization in two different ways. First, it increases the tendency of refolded, monomeric GroEL to undergo self-association at equilibrium. Second, crowding can modify the relative rates of the two competing self-association reactions, namely, productive assembly into a native tetradecameric structure and unproductive aggregation. This kinetic effect is most likely exerted by modifications of the diffusion coefficient of the refolded monomers, which in turn determine the conformational properties of the interacting subunits. If they are allowed to become assembly-competent before self-association, productive oligomerization occurs; otherwise, unproductive aggregation takes place. Our data demonstrate that the spontaneous refolding and assembly of homo-oligomeric proteins, such as GroEL, can occur efficiently (70%) under crowding conditions similar to those expected in vivo. PMID- 11020388 TI - Phosphorylation of p300 at serine 89 by protein kinase C. AB - CREB-binding protein (CBP)/p300 plays an important role in the connection of many different signal transduction pathways and the promotion of certain differentiation and proliferation processes. This role depends upon the ability of CBP/p300 to serve as coactivator for transcription factors. It has been suggested that CBP/p300 is regulated by phosphorylation, but the nature of the phosphorylation, the responsible kinase in vivo, and its physiological significance are still unclear. Here, we demonstrate the first identification of an in vivo phosphorylation site, conserved serine 89, in p300. Signal-dependent protein kinase C is able to phosphorylate serine 89 and mediates this phosphorylation event in vivo. Different from other phosphorylation observed so far in CBP/p300, this serine 89-specific phosphorylation represses the transcriptional activity of p300. This phosphorylation-mediated regulation of p300 function represents a previously unrecognized signal transduction pathway for protein kinase C to regulate cell growth and differentiation. PMID- 11020387 TI - The overall conformation of conventional kinesins studied by small angle X-ray and neutron scattering. AB - The quaternary structures of several monomeric and dimeric kinesin constructs from Homo sapiens and Drosophila melanogaster were analyzed using small angle x ray and neutron scattering. The experimental scattering curves of these proteins were compared with simulated scattering curves calculated from available crystallographic coordinates. These comparisons indicate that the overall conformations of the solution structures of D. melanogaster and H. sapiens kinesin heavy chain dimers are compatible with the crystal structure of dimeric kinesin from Rattus norvegicus. This suggests that the unusual asymmetric conformation of dimeric kinesin in the microtubule-independent ADP state is likely to be a general feature of the kinesin heavy chain subfamily. An intermediate length Drosophila construct (365 residues) is mostly monomeric at low protein concentration whereas at higher concentrations it is dimeric with a tendency to form higher oligomers. PMID- 11020390 TI - The incidence of pulmonary embolism in unexplained sudden cardiac arrest with pulseless electrical activity. AB - PURPOSE: The cause of many cases of sudden cardiac arrest from pulseless electrical activity is unknown. We hypothesized that pulmonary embolism was responsible for a substantial proportion of these cases and used transesophageal echocardiography to identify pulmonary embolism among patients with sudden cardiac arrest. SUBJECTS AND METHODS: We performed a prospective study at a tertiary care, university-operated county hospital, with a level 1 trauma center. Consecutive patients (n = 36) who were admitted with (n = 20) or unexpectedly developed (n = 16) sudden cardiac arrest of unknown cause were studied with transesophageal echocardiography during cardiopulmonary resuscitation. We determined the presence of central pulmonary embolism, right ventricular enlargement, and other causes of sudden cardiac arrest (such as myocardial infarction and aortic dissection) using prospectively defined criteria. RESULTS: Of the 25 patients with pulseless electrical activity as the initial event, 9 (36%) had pulmonary emboli (8 seen with transesophageal echocardiography and 1 diagnosed at autopsy) compared with none of the 11 patients with other rhythms, such as asystole or ventricular tachycardia or fibrillation (P = 0.02). Of the 8 patients who had pulmonary embolism diagnosed by transesophageal echocardiography, 2 survived to hospital discharge. CONCLUSIONS: Mortality from massive pulmonary embolism is high, particularly if patients present with sudden cardiac arrest. Earlier diagnosis of pulmonary embolus may permit wider use of thrombolytic agents or other interventions and may potentially increase survival. PMID- 11020389 TI - Rapid kinetic studies link tetrahydrobiopterin radical formation to heme-dioxy reduction and arginine hydroxylation in inducible nitric-oxide synthase. AB - To understand how heme and (6R)-5,6,7,8-tetrahydro-l-biopterin (H(4)B) participate in nitric-oxide synthesis, we followed ferrous-dioxy heme (Fe(II)O(2)) formation and disappearance, H(4)B radical formation, and Arg hydroxylation during a single catalytic turnover by the inducible nitric-oxide synthase oxygenase domain (iNOSoxy). In all cases, prereduced (ferrous) enzyme was rapidly mixed with an O(2)-containing buffer to start the reaction. A ferrous dioxy intermediate formed quickly (53 s(-1)) and then decayed with concurrent buildup of ferric iNOSoxy. The buildup of the ferrous-dioxy intermediate preceded both H(4)B radical formation and Arg hydroxylation. However, the rate of ferrous dioxy decay (12 s(-1)) was equivalent to the rate of H(4)B radical formation (11 s(-1)) and the rate of Arg hydroxylation (9 s(-1)). Practically all bound H(4)B was oxidized to a radical during the reaction and was associated with hydroxylation of 0.6 mol of Arg/mol of heme. In dihydrobiopterin-containing iNOSoxy, ferrous-dioxy decay was much slower and was not associated with Arg hydroxylation. These results establish kinetic and quantitative links among ferrous-dioxy disappearance, H(4)B oxidation, and Arg hydroxylation and suggest a mechanism whereby H(4)B transfers an electron to the ferrous-dioxy intermediate to enable the formation of a heme-based oxidant that rapidly hydroxylates Arg. PMID- 11020391 TI - Effects of age on the performance of common diagnostic tests for pulmonary embolism. AB - PURPOSE: The diagnosis of pulmonary embolism in the elderly is often difficult because of comorbid medical conditions, and perhaps also because diagnostic tests have a lower yield. We analyzed the diagnostic performance of common diagnostic tests for pulmonary embolism in different age groups. METHODS: We analyzed data from two large studies that enrolled 1,029 consecutive patients presenting to the emergency department with clinically suspected pulmonary embolism. The clinical probability of pulmonary embolism (high [>/=80%], intermediate, or low [/=80 years of age. The positive predictive value of a high clinical probability of pulmonary embolism was greater in the elderly (71% to 78% in those >/=60 years old versus 40% to 64% in those /=80 years old. The diagnostic yield of lower limb compression ultrasonography was greater in the elderly. The proportion of lung scans that were diagnostic (normal, near-normal, or high probability) decreased from 68% to 42% with increasing age. CONCLUSIONS: Age affects the performance of common diagnostic tests for pulmonary embolism and should be kept in mind when evaluating patients suspected of having this condition. PMID- 11020392 TI - Hydrochlorothiazide reduces loss of cortical bone in normal postmenopausal women: a randomized controlled trial. AB - PURPOSE: Thiazide diuretics reduce urine calcium excretion and might therefore reduce postmenopausal bone loss. In some, but not all, case-control studies, their use has been associated with a reduced incidence of hip fractures. We studied the effects of hydrochlorothiazide on bone loss in normal postmenopausal women. SUBJECTS AND METHODS: We performed a randomized, double-blind, 2-year trial of the effects of hydrochlorothiazide (50 mg per day) and placebo on bone mineral density in normal postmenopausal women. Participants were not required to have either low bone mineral density or hypertension. Bone mineral density was measured using dual-energy x-ray absorptiometry. RESULTS: One hundred eighty-five women entered the study, of whom 138 completed 2 years of follow-up. In an intention-to-treat analysis, hydrochlorothiazide produced significant benefits on bone mineral density of the total body (between-group difference at 2 years of 0.8%, 95% confidence interval [CI]: 0.3% to 1.3%, P <0.0001), legs (0.9%, 95% CI: 0.2% to 1.7%, P <0.0001), mid-forearm (1.2%, 95% CI: 0.2% to 2.2%, P = 0.02), and ultradistal forearm (1.7%, 95% CI: 0.1% to 3.2%, P = 0.04). There was no effect in the lumbar spine (0.5%, 95% CI: -0.5% to 1.6%) or femoral neck (0.2%, 95% CI: 1.3% to 1.7%). The between-group changes tended to be greatest during the first 6 months, except in the mid-forearm where there appeared to be a progressive divergence. An as-treated analysis produced similar results. Urine calcium excretion and indices of bone turnover decreased in the thiazide group, but parathyroid hormone concentrations did not differ between the groups. Treatment was tolerated well. CONCLUSIONS: Hydrochlorothiazide (50 mg per day) slows cortical bone loss in normal postmenopausal women. It may act directly on bone as well as on the renal tubule. The small size of the effect suggests that thiazides may have a role in the prevention of postmenopausal bone loss, but that they are not an appropriate monotherapy for treating osteoporosis. PMID- 11020393 TI - Evaluation of the effects of aspirin combined with angiotensin-converting enzyme inhibitors in patients with coronary artery disease. AB - BACKGROUND: Several studies have suggested that there may be an interaction between angiotensin-converting enzyme (ACE) inhibitors and aspirin in patients with congestive heart failure, such that their benefits are attenuated when used in combination. Whether this interaction exists in patients with coronary artery disease is not known. SUBJECTS AND METHODS: Patients enrolled in two large trials, Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO-I) and Evaluation in PTCA to Improve Long-Term Outcome with Abciximab GP IIb/IIIa Blockade (EPILOG), were stratified according to use of aspirin and ACE inhibitors on discharge from the hospital. In the EPILOG trial, left ventricular systolic function was assessed by contrast ventriculography. The primary endpoint was all-cause mortality at 1 year. EPILOG patients, all of whom were receiving aspirin, were also examined for the combined endpoint of death or nonfatal myocardial infarction. Stratified and multivariate analyses were used to adjust for baseline differences in patient characteristics. RESULTS: We studied 31,622 patients in the GUSTO-I trial and 2,619 patients in the EPILOG trial. There were 615 deaths among the GUSTO-I patients and 45 deaths among the EPILOG patients at 1 year. Unadjusted mortality was greater among patients treated with both ACE inhibitors and aspirin than among patients treated with aspirin alone (3.3% versus 1.6%, P <0.001 for GUSTO-I; and 3.7% versus 1.2%, P <0.001 for EPILOG). Similarly, the composite endpoint of death or nonfatal myocardial infarction was more frequent among EPILOG patients who were taking ACE inhibitors (6.3% versus 3.3%, P = 0. 001). After adjusting for confounders, combined use of aspirin and ACE inhibitors was associated with increased mortality in GUSTO-I patients (hazard ratio [HR] = 2.2, 95% confidence interval [CI]: 1.1 to 4.3, P = 0.03) compared with aspirin alone. In EPILOG patients, after adjusting for clinical factors and extent of left ventricular dysfunction, the combination of aspirin and ACE inhibitors was associated with an increased risk of death (HR = 2.1, 95% CI: 1.1 to 3.8, P = 0.02) and of death or nonfatal myocardial infarction (HR = 1.5, 95% CI: 1.1 to 2.5, P = 0.02) compared with aspirin alone. CONCLUSION: These observational findings suggest the possibility of an interaction between aspirin and ACE inhibitors among patients with ischemic heart disease. Further study of this issue is warranted. PMID- 11020394 TI - Relation between length of hospital stay and costs of care for patients with community-acquired pneumonia. AB - PURPOSE: Patients with pneumonia often remain hospitalized after becoming clinically stable, without demonstrated benefits on outcome. The purposes of this study were to assess the relation between length of hospital stay and daily medical care costs and to estimate the potential cost savings associated with a reduced length of stay for patients with pneumonia. SUBJECTS AND METHODS: As part of a prospective study of adults hospitalized with community-acquired pneumonia at a community hospital and two university teaching hospitals, daily medical care costs were estimated by multiplying individual charges by department-specific cost-to-charge ratios obtained from each hospital's Medicare cost reports. RESULTS: The median total cost of hospitalization for all 982 inpatients was $5, 942, with a median daily cost of $836, including $491 (59%) for room and $345 (41%) for non-room costs. Average daily non-room costs were 282% greater on the first hospital day, 59% greater on the second day, and 19% greater on the third day than the average daily cost throughout the hospitalization (all P <0.05), and were 14% to 72% lower on the last 3 days of hospitalization. Average daily room costs remained relatively constant throughout the hospital stay, with the exception of the day of discharge. A projected mean savings of $680 was associated with a 1-day reduction in length of stay. CONCLUSIONS: Despite institutional differences in total costs, patterns of daily resource use throughout hospitalization were similar at all institutions. A 1-day reduction in length of stay might yield substantial cost-savings. PMID- 11020395 TI - Unplanned readmissions of patients with congestive heart failure: do they reflect in-hospital quality of care or patient characteristics? AB - PURPOSE: To determine if early unplanned readmissions of patients hospitalized for heart failure are associated with suboptimal in-hospital care or with the clinical and demographic characteristics of the patient. SUBJECT AND METHODS: We performed a case-control study among patients discharged with a principal diagnosis of heart failure. Cases included all patients unexpectedly readmitted within 31 days of discharge; controls were randomly selected from among those not readmitted. Quality of care was measured using explicit criteria reflecting the admission work-up, evaluation and treatment, and readiness for discharge. RESULTS: Ninety-one cases and 351 controls were included. There was no significant association between early unplanned readmissions and the scores for quality of the admission work-up or evaluation and treatment during the stay. There was a significant association between readiness for discharge and subsequent early readmission: for each 10% decrease in the proportion of fulfilled criteria, the odds of readmission increased by 14% (95% confidence interval [CI] 1. 01 to 1.28, P = 0.04) for all-cause readmissions and by 19% (95% CI: 1.04 to 1.36, P = 0.01) for heart-failure-related readmissions. In a multiple logistic regression model, previous diagnosis of heart failure (odds ratio [OR] = 2.9, 95% CI: 1.7 to 4.8, P <0.001), age (OR = 3.3, 95% CI: 1.3 to 8.5, P = 0.01 for patients aged 65 to 79 years and OR = 4.1, 95% CI: 1.6 to 11, P = 0.004 for patients aged 80 years and older), and history of cardiac revascularization (OR = 2.1, 95% CI: 1.2 to 3.9, P = 0.01) showed a stronger association with early unplanned all-cause readmissions than the readiness-for-discharge score (OR = 1.16, 95% CI: 1.02 to 1.31, P = 0.02). Similar findings were seen for heart failure-related readmissions. CONCLUSIONS: Among patients with heart failure, early unplanned readmissions were not associated with suboptimal admission work up or evaluation and treatment but were weakly associated with readiness for discharge. However, they were strongly associated with the patients' clinical and demographic characteristics. PMID- 11020396 TI - Hellenic holocaust: a historical clinico-pathologic conference. PMID- 11020397 TI - Oxidants, nitrosants, and the lung. AB - The respiratory tract is subjected to a variety of environmental stresses, including oxidizing gases, particulates, and airborne microorganisms, that together, may injure structural and functional lung components and thereby jeopardize the primary lung function of gas exchange. To cope with such various environmental threats, the lung has developed elaborate defense mechanisms that include inflammatory-immune pathways as well as several antioxidant systems. These defense systems operate largely in extracellular spaces, thus protecting underlying bronchial and alveolar epithelial cells from injury, although these cells themselves are also active participants in such (inflammatory) defense mechanisms. Although potentially harmful, oxidants are increasingly recognized as pathophysiologic mediators produced primarily by inflammatory-immune cells as a host defense mechanism, but also by various other cell types as an intracellular mediator in various cell responses, thus affecting inflammatory-immune processes or inducing resistance. The molecular mechanisms and signaling pathways involved in such processes are the focus of much current investigation. Nitric oxide, a messenger molecule produced by many lung cell types, also modulates oxidant mediated processes, thereby giving rise to a new family of reactive nitrogen species ("nitrosants") with potentially unique signaling properties. The complex role of oxidants and nitrosants in various pathophysiologic processes in the lung have confounded the design of therapeutic approaches with antioxidant substrates. This review discusses current knowledge regarding extracellular antioxidant defenses in the lung, and oxidant/nitrosant mechanisms operating under inflammatory-immune conditions and their potential contribution to common lung diseases. Finally, some recent developments in antioxidant therapeutic strategies are discussed. PMID- 11020398 TI - Tropical pyomyositis and human toxocariasis: a clinical and experimental study. PMID- 11020399 TI - Pulmonary embolism: a treatable cause of cardiac arrest? PMID- 11020400 TI - Thiazides and bone. PMID- 11020401 TI - Interaction between aspirin and angiotensin-converting enzyme inhibitors: real or imagined. PMID- 11020402 TI - How long a stay in the hospital is needed for patients with community-acquired pneumonia? PMID- 11020403 TI - Ahmed glaucoma valve implant vs trabeculectomy in the surgical treatment of glaucoma: a randomized clinical trial. AB - PURPOSE: To compare the short- and intermediate-term results of two commonly used glaucoma surgical procedures, trabeculectomy and Ahmed glaucoma valve implant. METHODS: A randomized clinical trial was performed at two international centers. One eye each of consecutive patients requiring glaucoma surgery for intraocular pressure control was randomized to receive either trabeculectomy or the Ahmed implant. RESULTS: Of the 117 patients, 62 were randomized to trabeculectomy and 55 to the Ahmed implant. With a mean follow-up of 9.7 months, the trabeculectomy group had statistically lower intraocular pressures at weeks 6 to 15 (12.6 mm Hg vs 16.4 mm Hg) and months 11 to 13 (11.4 mm Hg vs 17.2 mm Hg) than the Ahmed implant group. Compared with preoperative status, no statistically significant differences between groups were noted for visual acuity, visual field, lens status, and final anterior chamber depth. The cumulative probabilities of success (intraocular pressure <21 mm Hg and at least 15% reduction in intraocular pressure from preoperative level) were 83.6% for trabeculectomy and 88.1% for Ahmed implant (P =.43). However, the Ahmed implant group had a greater adjunctive medication requirement. On the last visit, 10 of the trabeculectomy eyes and 19 of the Ahmed implant eyes required at least one topical medication (P =.01). There was no statistically significant difference in the rate of complications between the two groups. CONCLUSIONS: Lower mean intraocular pressures were noted for the trabeculectomy group. All other results, including success (as defined in this study) and frequency of complications, were comparable between the two groups. PMID- 11020404 TI - Factors associated with long-term progression or stability in primary open-angle glaucoma. AB - PURPOSE: To evaluate long-term risk factors for progression or stability in patients with primary open-angle glaucoma. METHOD: We retrospectively included consecutively reviewed patients who had primary open-angle glaucoma for at least 5 years in this multicenter trial. Historical and clinical factors in these patients were evaluated for their association with stability or progression of the glaucoma. RESULTS: We included 218 patients in this study; of these, 34 progressed over an average length of follow-up of 45.5 +/- 30.0 months, and 184 were stable over an average of 72.8 +/- 18.3 months. The mean intraocular pressure over the follow-up period for the progressed group was 19.5 +/- 3.8 mm Hg and for the stable group 17. 2 +/- 3.1 mm Hg (P =.001). The average standard deviation of individual intraocular pressures was greater in the progressed group (5.1 mm Hg) than the stable group (3.9 mm Hg, P =.012). Baseline characteristics indicating a greater potential to progress were a larger cup-to-disk ratio (P <.001), a greater number of medications (P =.02), older age (P.007), and worse visual acuity (P =.003). However, no difference was observed in pressure levels that prevented progression in these subpopulations compared with the total sample size. CONCLUSIONS: This study suggests that lowering the intraocular pressure is important in the treatment of primary open-angle glaucoma to help prevent long term progression. Lowering the pressure, however, is not uniformly effective in preventing progression. Additionally, risk factors for progression do not further help identify pressure levels that prevent worsening of glaucoma. PMID- 11020405 TI - Optic disk topography after medical treatment to reduce intraocular pressure. AB - PURPOSE: We examined changes in optic disk topography using confocal scanning laser ophthalmoscopy after reducing intraocular pressure with administration of latanoprost. METHODS: Twenty-nine patients with glaucoma or ocular hypertension were imaged using the Heidelberg Retina Tomograph before and after the administration of latanoprost to decrease intraocular pressure. Average time between pretreatment and posttreatment imaging was 2.7 +/- 1.8 weeks. Heidelberg Retina Tomograph software-measured parameters were mean height of contour, cup area, cup volume, mean cup depth, maximum cup depth, cup shape, rim area, rim volume, cup-to-disk ratio, and retinal nerve fiber thickness. RESULTS: Average intraocular pressure decreased significantly (mean +/- SD) by 7.2 +/- 5.4 mm Hg (25 +/- 16% decrease). No statistically significant changes in measured topographic parameters were found. When data from patients with decreases in intraocular pressure of 7 mm Hg or greater were analyzed separately (mean intraocular pressure decrease = 10.79 +/- 4.32 mm Hg, 36 +/- 8% decrease), cup area (P =.005), cup volume (P =. 002), and cup-to-disk ratio (P =.005) decreased significantly, and rim area (P =.005) increased significantly. Linear regression analysis of the data from all subjects showed that a change in intraocular pressure after latanoprost administration accounted for 12% or more of the variance in two measured topographic parameters (mean cup depth and cup shape). CONCLUSIONS: These results suggest that, in some patients, moderate decreases in intraocular pressure may affect disk topography, as measured by Heidelberg Retina Tomograph. Intraocular pressure should be considered when analyzing consecutive confocal scanning laser ophthalmoscopy images for glaucomatous progression. PMID- 11020406 TI - Anterior uveitis as a side effect of topical brimonidine. AB - PURPOSE: To report anterior uveitis as a late side-effect of topical brimonidine tartrate 0.2% (Alphagan; Allergan Inc., Irvine, California). METHODS: Four patients aged 79 to 91 (mean, 83) years, who developed acute anterior uveitis while using topical brimonidine for chronic glaucoma, were clinically evaluated for causes of uveitis. When the inflammation had completely cleared, topical brimonidine was recommenced in one eye only to determine causality. RESULTS: All four patients developed acute granulomatous anterior uveitis, which resolved rapidly on stopping brimonidine and receiving treatment with topical corticosteroids. The uveitis occurred 11 to 15 (mean, 13.8) months after commencement of brimonidine 0.2% twice daily. None of the patients had a previous history of uveitis, and no other identifiable cause of uveitis was found. In all patients, on rechallenge with brimonidine instilled twice daily to one eye, uveitis recurred within 3 weeks in that eye only. CONCLUSIONS: These findings suggest that brimonidine can cause anterior uveitis as a late side effect. The inflammation settles rapidly on stopping the medication and on using topical corticosteroids and recurs on rechallenge with brimonidine. PMID- 11020407 TI - Phakic posterior chamber intraocular lens for the correction of anisometropia and treatment of amblyopia. AB - PURPOSE: To assess the potential visual benefits of posterior chamber phakic intraocular lens implants in eyes of children with anisometropic amblyopia. METHODS: In a prospective study, three girls 9, 14, and 18 years old with high anisometropia and deep amblyopia were included in this study. The phakic posterior chamber intraocular lens (ICL; STAAR Surgical AG, Nidau, Switzerland) was used to correct the anisometropia. This intraocular lens was inserted in the anterior chamber through a 3.0-mm temporal clear cornea incision and manipulated into the posterior chamber using an iris manipulator. A peripheral iridectomy was performed using the Ocutome Probe (Storz; Premiere, St. Louis, Missouri). Local therapy with corticosteroids and antibiotics were prescribed for 2 weeks, and patients were followed regularly for a period of 6 to 9 months. RESULTS: In the three amblyopic eyes of the three patients, the preoperative best-corrected visual acuity of 6/30, 6/60, and 6/30 improved, to 6/7.5 (20/25), 6/30 (20/100), and 6/15 (20/50), respectively, 6 months after the surgery. Binocular functions with development of fusional abilities and stereopsis were observed in two of these patients after the intraocular lens implantation. In the third patient, the fusional abilities developed only after surgical correction of the exotropia. The intraocular pressure remained within normal limits, and there was no significant change in the corneal endothelial cell count during the period of follow-up. No major intraoperative or postoperative complications were observed, except for a temporary pigment dispersion. CONCLUSIONS: Implantation of phakic posterior chamber intraocular lenses may be beneficial for the treatment of amblyopia in children with anisometropia. Although additional cases and long-term follow-up observations are necessary, it appears that amblyopia may be overcome by the use of posterior chamber phakic intraocular lens implants, even in eyes of children beyond the age generally considered to be responsive to anti-amblyopic treatment. PMID- 11020408 TI - Sloughing of corneal epithelium and wound healing complications associated with laser in situ keratomileusis in patients with epithelial basement membrane dystrophy. AB - PURPOSE: To report sloughing of corneal epithelium during laser in situ keratomileusis and subsequent wound healing complications in patients with epithelial basement membrane dystrophy. METHODS: In a retrospective study, the surgical procedures, postoperative course, and visual acuities of 16 eyes of nine patients with epithelial basement membrane dystrophy who underwent laser in situ keratomileusis complicated with epithelial sloughing at three centers were reviewed. The mean follow-up period was 23 weeks (range, 4 to 52 weeks). RESULTS: In 13 (81%) of 16 eyes with epithelial basement membrane dystrophy, epithelial sloughing occurred during laser in situ keratomileusis. In eight of the 13 eyes, epithelial growth beneath the flap was observed. The flap was lifted and the interface epithelium scraped in six eyes. Flap melt or keratolysis occurred in four eyes. At the last follow-up visit, 13 of 16 eyes had an uncorrected visual acuity of 20/30 or better, and all eyes had a best-corrected visual acuity of 20/30 or better. CONCLUSIONS: Patients with epithelial basement membrane dystrophy have poorly adherent corneal epithelium and are predisposed to epithelial sloughing during the microkeratome pass of laser in situ keratomileusis. This may lead to flap distortion, interface epithelial growth, flap keratolysis, and corneal scarring. It is not recommended that laser in situ keratomileusis be performed in patients with classic, symptomatic epithelial basement membrane dystrophy. In patients who present with mild and asymptomatic epithelial basement membrane dystrophy, laser in situ keratomileusis should be performed with caution, or photorefractive keratectomy may be the preferred refractive procedure. PMID- 11020409 TI - The madurai intraocular lens study IV: posterior capsule opacification. AB - PURPOSE: To estimate the cumulative incidence of posterior capsule opacification 4 years after surgery in patients who participated in the Madurai Intraocular Lens Study and had extracapsular cataract extraction with posterior chamber intraocular lens implantation. METHODS: In the Madurai Intraocular Lens Study, 1,700 patients with best-corrected visual acuity 20/120 or worse in the better eye had extracapsular cataract extraction with posterior chamber intraocular lens implantation, and 1,474 (86.7%) of these completed the 1-year follow-up examination. From this group of 1,474 pseudophakic patients, 400 were randomly selected for reexamination 4 years after the original surgery. The eye that was operated on was examined by an ophthalmologist who was involved in the 1-year follow-up examinations and posterior capsule opacification grading. A grading of I to III was used to reflect the degree of opacification. With grades II and III, posterior capsule opacification detectable with an undilated pupil was present in the central axis. RESULTS: Three hundred twenty-seven (81.8%) of the selected population were examined between October 1997 and December 1998. Thirty-four (8.5%) were confirmed as being deceased, and 39 (9.8%) were unavailable for follow-up. The median age was 60 years, and 57.2% were women. The 4-year incidence of grade II or III posterior capsule opacification, including eyes already treated with laser capsulotomy, was 13.1% (95% confidence interval [CI], 9.7% to 17.3%). Each year of increased age was associated with a decreased risk of posterior capsule opacification (odds ratio, 0.96; 95% CI, 0.92 to 1.00). Based on best-corrected visual acuity of 20/40 or worse without co-existing pathology, the 4-year incidence of posterior capsule opacification was 13.5%. CONCLUSION: Because patients with relatively mature cataracts routinely receive extracapsular cataract extraction with posterior chamber intraocular lens implantation instead of the traditional intracapsular extraction, the subsequent need for laser capsulotomy may be less than that anticipated, based on previous reports. PMID- 11020410 TI - Clinical assessment of long-term safety and efficacy of a widely implanted polyacrylic intraocular lens material. AB - PURPOSE: To evaluate the long-term safety and efficacy, in a large series of patients, of polyacrylic intraocular lens implants 1 and 3 years after surgery, compared with polymethylmethacrylate intraocular lens implants. METHODS: This was a randomized, prospective study of 1,514 eyes of 1,202 patients that were examined for a 1-year follow-up study and 1,264 eyes of 990 patients that were examined for a 3-year follow-up study. All the patients were prospectively randomized to receive a polyacrylic or polymethylmethacrylate intraocular lens implant. A standardized surgical protocol was performed using a phacoemulsification technique and capsulorhexis. Any surgical complications were excluded, and all patients had standard postoperative medication and follow-up. Postoperative measurements included spectacle-corrected visual acuity, occurrence of postoperative sight-threatening or lens-related complications, and adverse reactions. Results were compared between the polyacrylic intraocular lens and polymethylmethacrylate intraocular lens groups. Results were also compared with standards for polymethylmethacrylate intraocular lens that were established by the US Food and Drug Administration and a previous study for silicone lens. RESULTS: The overall complete follow-up rates were 79.9% for 1-year follow-up and 69.5% for 3-year follow-up. At 1 year, the mean logarithm of the minimum angle of resolution (logMAR) of visual acuity was 0.037 +/- 0.150 and 0.042 +/- 0.154 in best case patients of the polyacrylic intraocular lens group and polymethylmethacrylate intraocular lens group, respectively. At 3 years, the mean logMAR visual acuity was 0.038 +/- 0.155 and 0.054 +/- 0.181 in best case patients in the polyacrylic intraocular lens group and polymethylmethacrylate intraocular lens group. There was no significant difference between groups at 1 and 3 years. At 1 year, 96.8% of best case patients in the polyacrylic intraocular lens group and 95.8% of those in the polymethylmethacrylate intraocular lens group achieved corrected visual acuity of 0.5 or better. At 3 years, 96.4% of best case patients in the polyacrylic intraocular lens group and 94.9% of those in the polymethylmethacrylate group achieved a corrected visual acuity of 0.5 or better; no significant difference was found at 1 and 3 years. The rate of sight-threatening complications was 1. 2% for the polyacrylic intraocular lens group of all patients and 1. 5% for the polymethylmethacrylate intraocular lens group at 1 year, and 1.6% for the polyacrylic intraocular lens group and 1.4% for the polymethylmethacrylate intraocular lens group at 3 years. No significant difference was found between the groups. The cumulative rate of Nd:YAG capsulotomy for the polyacrylic intraocular lens group (7.1%) was significantly lower than that for the polymethylmethacrylate intraocular lens group (38.1%) through 3 years (P <.0001). CONCLUSIONS: Polyacrylic intraocular lenses demonstrated safe and effective performance through long-term follow-up at a level equal to or better than polymethylmethacrylate intraocular lenses. Patients undergoing polyacrylic intraocular lens implantation achieved excellent postoperative visual acuity and experienced a low rate of postoperative complications, adverse reactions, and Nd:YAG capsulotomy. These long-term clinical results are an important outcome database for the ongoing use of this foldable optic material. PMID- 11020411 TI - Reproductive exposures, incident age-related cataracts, and age-related maculopathy in women: the beaver dam eye study. AB - PURPOSE: To evaluate the relationship of reproductive exposures and incident age related cataract and maculopathy in women. METHODS: This was a population-based cohort study including all adults 43 to 84 years of age living in Beaver Dam, Wisconsin (a representative midwestern community) who were identified during a census in 1987 to 1988. They were evaluated initially in 1988 to 1990 and at follow-up in 1993 to 1995. Evaluations included medical histories and both fundus and lens photography. All procedures were done according to protocols that were the same at both examinations. All photographs were graded by trained observers using defined grading schemes. The severities of age-related cataracts and maculopathy were determined by grading of photographs. Information on hormone exposures was ascertained from structured interviews. RESULTS: After adjusting for age, the only significant finding for lens end points was a trend indicating a possible protective effect of increasing number of live births and incident posterior subcapsular cataract. There were no significant associations of any reproductive exposure with lesions of early or late age-related maculopathy. CONCLUSIONS: In these population-based data, there is little evidence of association of hormone exposures with incident age-related eye disease in women 5 years later. Longer follow-up of this population, whose mean age is approaching that of heightened incidence, may disclose significant relationships. PMID- 11020412 TI - Optic disk edema associated with peripapillary serous retinal detachment: an early sign of systemic Bartonella henselae infection. AB - PURPOSE: To describe optic disk edema associated with peripapillary serous retinal detachment as an early sign of systemic Bartonella henselae infection. METHODS: Multicentered, retrospective case series. RESULTS: Five women and two men presented with optic disk edema producing peripapillary serous retinal detachment. Each patient had a markedly elevated serum anti-B. henselae antibody titer. Patient age ranged from 11 to 44 years, with a mean and median of 26.6 and 28 years, respectively. The time from the onset of systemic symptoms to the onset of visual symptoms varied from 3 days to 1 month. The peripapillary serous retinal detachment resolved within 1 to 3 weeks in each case, producing a macular star in four of seven patients. Initial vision was 20/200 or worse in five of seven patients and improved in four of these five patients to 20/30 or better. CONCLUSIONS: Systemic B. henselae infection should be considered in patients who develop optic disk edema associated with a peripapillary serous retinal detachment, even in the absence of classic neuroretinitis with a macular star. PMID- 11020413 TI - Outcomes of macular hole surgery in severely myopic eyes: a case-control study. AB - PURPOSE: To evaluate the outcomes of macular hole surgery in highly myopic eyes and to compare these outcomes with a control group of eyes that were not severely myopic. METHODS: The study design was a matched, case-control, retrospective chart review. The participants included 26 eyes of 24 patients who had vitreous surgery for macular holes. The eyes were divided into two groups: 13 consecutive eyes with severe myopia (defined as -6.00 diopters of refractive error or greater) and 13 control eyes without severe myopia that were operated on immediately before or after each study eye, with the most recently operated eye chosen. The main outcome parameters were preoperative and final follow-up visual acuity, macular hole closure rates, reoperation rates, duration of preoperative symptoms, and follow-up time. RESULTS: Using the Snellen equivalent of logarithm of minimal angle of resolution (logMAR) units, visual acuity improved after macular hole surgery in severely myopic eyes from 20/152 to 20/89 (P =.041) and in control eyes from 20/152 to 20/47 (P <.001). At final follow-up, visual acuities were lower in severely myopic eyes compared with control eyes (P =.048). Macular hole closure rates, reoperation rates, duration of pre-operative symptoms, and follow-up intervals were not significantly different statistically between groups. CONCLUSIONS: Macular hole surgery results in anatomical and visual improvements in severely myopic eyes but generally yields poorer visual acuity outcomes compared with eyes that are not severely myopic. PMID- 11020414 TI - Ocular bartonellosis. AB - PURPOSE: To review recent advances in the basic and clinical biology of Bartonella-related eye disease. METHOD: A review of the pertinent medical literature was performed. RESULTS: A number of novel Bartonella species have been identified over the past decade. Of these, Bartonella henselae, the etiologic agent in cat scratch disease, is most often associated with ocular complications, which may include Parinaud oculoglandular syndrome, neuroretinitis, and focal retinochoroiditis. Although cat and flea exposure appear to be the main risk factors for contracting cat scratch disease, the diagnosis of ocular bartonellosis relies primarily on the recognition of suggestive clinical signs in conjunction with positive serologic testing. B. henselae-associated ocular complications are usually self-limited but may be treated with doxycycline or erythromycin, with or without rifampin, when the infections are severe or sight threatening. CONCLUSIONS: B. henselae infection is common and should be considered in patients with Parinaud oculoglandular syndrome, neuroretinitis, or focal retinochoroiditis, particularly when there is a history of cat or flea exposure. PMID- 11020415 TI - A short questionnaire on visual function of older adults to supplement ophthalmic examination. AB - PURPOSE: To estimate frequency of self-reported visual dysfunction in specific life situations. METHODS: Performance-based measures of visual function were obtained during the 5-year follow-up examination of the Beaver Dam Eye Study cohort. During the subsequent yearly follow-up telephone call, four questions concerning visual abilities in specific situations were administered. RESULTS: Visual difficulties in everyday life situations were commonly reported by adults aged 48 to 91 years, with only 17% to 35% of persons reporting excellent or very good vision in the four specific situations related to visual function. Visual acuity, near vision, and contrast sensitivity measures do not reflect the visual difficulties encountered in common daily activities in our study population (Spearman correlation coefficients all less than 0.35). CONCLUSIONS: A few simple questions about visual function in common daily activities may give a clinician insights into patient complaints. It may be that environmental changes could improve self-reported visual function. PMID- 11020416 TI - Epilenticular intraocular lens implantation in traumatic cataract with a ruptured posterior capsule. AB - PURPOSE: To present a case of rapidly progressing traumatic cataract caused by posterior capsule rupture after nonpenetrating ocular injury, as well as a surgical procedure to safely implant the intraocular lens in such a traumatized eye. METHODS: In a 23-year-old man with traumatic cataract and posterior lens capsule rupture, a one-piece polymethylmethacrylate IOL was implanted before cataract extraction into the ciliary sulcus in front of the cataractous lens. Subsequently, the cataract was removed by pars plana lensectomy. RESULTS: The surgery and postoperative course were uneventful. Postoperative visual acuity was 20/20. CONCLUSION: We present a method of intraocular lens implantation in cases of rapidly progressing traumatic cataract caused by posterior capsule rupture after a blunt ocular trauma. PMID- 11020417 TI - Influence of rapid glycemic control on lens opacity in patients with diabetes mellitus. AB - PURPOSE: To report the influence of rapid glycemic control on lens opacity in patients with diabetes mellitus. METHODS: In a prospective study, nine patients with adult onset diabetes mellitus and glycosylated hemoglobin values over 9% were divided into two groups, rapid glycemic control and slow glycemic control groups, based on the time course of glycosylated hemoglobin values after the initiation of glycemic control. The lens thickness and opacity were measured using the anterior eye segment analysis system. RESULTS: One week after onset of treatment, the lens in rapid glycemic control group became significantly thicker than in pretreatment, but returned to the baseline level at the subsequent measurement points. The lens opacity index in the rapid glycemic control group increased significantly (P <.01, paired t test) 4 months after the glycemic control, which persisted throughout the 1-year study period. The lens thickness and opacity in the slow glycemic control group did not change significantly. CONCLUSION: It was suggested that rapid glycemic control can induce an irreversible increase in lens opacification. PMID- 11020418 TI - Amsler grid examination and optical coherence tomography of a macular hole caused by accidental Nd:YAG laser injury. AB - PURPOSE: To compare a macular hole from accidental Nd:YAG laser injury with idiopathic macular holes. METHODS: Case report. In a 24-year-old man with accidental Nd:YAG laser injury, right eye, Amsler grid testing and optical coherence tomography were performed. RESULTS: Nd:YAG laser injury was responsible for a macular hole about 700 microm in diameter. The visual acuity was 20/100. Amsler grid testing displayed a central scotoma with no surrounding distortion. Optical coherence tomography showed a defect in all retinal layers at the macula. CONCLUSION: The scotoma caused by Nd:YAG laser injury is not surrounded by distortion; the hole is produced by the defect of all retinal layers. In contrast, idiopathic macular holes generally produce a pincushion pattern on Amsler grid testing and have no tissue loss. PMID- 11020419 TI - Novel mutation in RP2 gene in two brothers with X-linked retinitis pigmentosa and mtDNA mutation of leber hereditary optic neuropathy who showed marked differences in clinical severity. AB - PURPOSE: To report the identification of a novel mutation of the RP2 gene in two Japanese brothers with X-linked retinitis pigmentosa of a differing clinical severity. The mother was a carrier of both retinitis pigmentosa and optic atrophy. METHODS: The older brother had a severe form of retinitis pigmentosa associated with macular degeneration and total optic atrophy, whereas the younger brother presented typical X-linked retinitis pigmentosa. RESULTS: Each patient exhibited a novel 2-bp insertion at codon 278 in exon 3 of the RP2 gene as well as a 11778 mutation in mitochondrial DNA. This suggests that the older brother may have developed Leber hereditary optic neuropathy as well as retinitis pigmentosa. CONCLUSION: Molecular testing confirmed the clinical diagnosis in each case. However, such testing did not explain the differences in the severity of the ophthalmoscopic findings between the two brothers. PMID- 11020420 TI - Mizuo phenomenon observed by scanning laser ophthalmoscopy in a patient with Oguchi disease. AB - PURPOSE: To elucidate the origin of the abnormal fundus reflex in Oguchi disease. METHODS: The ocular fundus of a 63-year-old woman who showed a homozygous arrestin 1147delA mutation was observed by scanning laser ophthalmoscopy with the use of an argon blue laser (wavelength, 488 nm), a helium-neon laser (633 nm), and an infrared laser (780 nm). RESULTS: Diffuse, fine, white particles, which do not exist in normal subjects, were clearly demonstrated only with the helium-neon laser. After 4-hour dark adaptation, the abnormal particles disappeared, but then they reappeared gradually during 30 minutes of light adaptation, in accordance with the golden metallic reflex. CONCLUSION: The white particles found by helium neon laser could be the origin of the abnormal fundus reflex in Oguchi disease. PMID- 11020421 TI - Acquired and progressive retinal nerve fiber layer myelination in an adolescent. AB - PURPOSE: To report the onset and progression of retinal nerve fiber layer myelination in an adolescent patient. METHODS: Case report. RESULTS: A 7-year-old male was found to have myelination of the retinal nerve fiber layer in the left eye. When he was reexamined at age 14 years, he was found to have new myelination in the right eye, progression of the myelination in the left eye, and bilateral optic nerve drusen. CONCLUSION: Retinal nerve fiber layer myelination can occur spontaneously in adolescence, progress in adolescence, and be associated with optic nerve drusen. PMID- 11020422 TI - Bilateral diffuse choroidal hemangiomas with unilateral facial nevus flammeus in Sturge-Weber syndrome. AB - PURPOSE: To report bilateral choroidal hemangiomas associated with unilateral facial nevus flammeus in Sturge-Weber syndrome. METHODS: Case report. RESULTS: A 6-year-old male with a left facial nevus flammeus and a history of a left cerebral angioma had been followed 2 years for increasing esotropia. Examination demonstrated bilateral diffuse choroidal hemangiomas with overlying exudative retinal detachments. After bilateral external beam radiotherapy, the retinal detachments resolved and vision improved. CONCLUSION: Patients with Sturge-Weber syndrome and unilateral facial nevus flammeus may harbor bilateral choroidal hemangiomas. Clinical manifestations of the Sturge-Weber syndrome are characteristically unilateral and ipsilateral to the facial nevus flammeus. Bilateral choroidal hemangiomas associated with bilateral facial nevus flammeus are rare. 1,2 We report bilateral diffuse choroidal hemangiomas associated with unilateral facial nevus flammeus in Sturge-Weber syndrome. PMID- 11020423 TI - Echographic features of medulloepithelioma. AB - PURPOSE: To evaluate the echographic features of medulloepithelioma that may assist in establishing the diagnosis. METHODS: Retrospective review identified four eyes with medulloepithelioma studied with echography. Clinical records, echographic findings, histopathologic tumor features, and the clinical course were reviewed. RESULTS: The initial preoperative diagnosis of medulloepithelioma was uncertain, based on clinical findings alone in three cases but was accurate when echographic findings were combined with clinical findings in all four cases. Cysts posterior to the iris were detected on clinical examination in only two cases, but were revealed on echography in all four cases. Additional echographic findings included irregular high internal reflectivity (n = 4), irregular tumor surface (n = 3), molding around intraocular structures (n = 3), and internal vascularity (n = 2). CONCLUSIONS: The diagnosis of medulloepithelioma is not always apparent on clinical examination alone. Echographic findings of a highly reflective, irregularly structured tumor with associated cystic changes involving the ciliary body region may help establish a presumed diagnosis of medulloepithelioma. PMID- 11020424 TI - Bilateral blindness and lumbosacral myelopathy associated with high-dose carmustine and cisplatin therapy. AB - PURPOSE: To report the early ocular pathologic findings associated with high-dose carmustine and cisplatin therapy. METHODS: A patient with metastatic breast carcinoma developed an acute onset of branch retinal artery occlusion, bilateral blindness, and a myelopathy involving the lower extremities after high-dose chemotherapy and bone marrow transplant. RESULTS: Histopathologic examination of the eye and optic nerves at autopsy disclosed nerve fiber layer infarction secondary to right inferior temporal retinal artery thrombosis. Patchy necrosis of both optic nerves, medulla oblongata, and spinal cord was associated with focal small-vessel thrombosis. CONCLUSIONS: The syndrome of retinal vascular occlusion, optic neuropathy, and myelopathy is associated with the high-dose chemotherapeutic agents carmustine and cisplatin. The distribution of necrosis suggests an ischemic event rather than direct neurotoxic effects. PMID- 11020425 TI - Osteogenesis in a human hydroxyapatite orbital implant 5.5 years after implantation. AB - PURPOSE: We report the histological findings of a hydroxyapatite orbital implant 5.5 years after implantation. METHODS: The hydroxyapatite orbital implant was excised from the left orbit of a 17-year-old female, and it was decalcified and processed for routine light microscopic evaluation. RESULTS: The implant showed complete fibrovascularization with nearly complete osteogenesis with hematopoiesis. CONCLUSION: Bone may biointegrate throughout the pores of hydroxyapatite orbital implants. PMID- 11020426 TI - Reversal of optic canal stenosis in osteopetrosis after bone marrow transplant. AB - PURPOSE: To describe a patient with infantile osteopetrosis and optic atrophy secondary to optic canal stenosis who demonstrated optic canal enlargement after bone marrow transplant. METHODS: Case report. A 3-month-old infant with infantile "malignant" osteopetrosis underwent ophthalmic examination, including visual evoked potentials, electroretinogram, and computed tomography (CT). Bone marrow transplant was performed at 8 months of age. RESULTS: Examination revealed visual loss and optic atrophy, left eye greater than right eye, secondary to optic canal stenosis. Flash visual evoked potentials revealed a normal waveform in both eyes with increased latency in the left eye. Electroretinogram was normal in both eyes. CT after bone marrow transplant showed enlargement of the optic canals. Vision remains stable 43 months after bone marrow transplant. CONCLUSIONS: Bone marrow transplant in infantile osteopetrosis may be followed by reversal of optic canal stenosis and preservation of vision. PMID- 11020427 TI - Orbital T-cell lymphoma in a multiple myeloma patient. AB - PURPOSE: To report an 82-year-old woman with multiple myeloma who developed an orbital T-cell lymphoma concomitantly. METHODS: The patient presented with left upper eyelid swelling. Magnetic resonance imaging demonstrated an orbital mass compressing the eyeball. The mass was excised for diagnostic purposes and orbital decompression. RESULTS: Histopathologic and immunohistochemical evaluation identified the mass as a T-cell lymphoma. The disease progressed rapidly despite chemotherapy. CONCLUSIONS: Orbital T-cell lymphoma is rare and we are unaware of previous reports of orbital T-cell lymphoma in patients with multiple myeloma. PMID- 11020428 TI - Recurrent orbital solitary fibrous tumor in a 14-year-old girl. AB - PURPOSE: To report a case of orbital solitary fibrous tumor in a pediatric patient. METHODS: Case report and review of the literature. RESULTS: A 14-year old girl presented with a 5-month history of painless proptosis of the left eye. Magnetic resonance imaging revealed a well-circumscribed mass in the anterior superomedial left orbit. The lesion was excised, and histopathologic examination revealed a solitary fibrous tumor. The lesion recurred in the orbit 4 months postoperatively, and histologic examination of the new lesion was consistent with solitary fibrous tumor. CONCLUSION: Based on this case report of orbital solitary fibrous tumor in a pediatric patient, solitary fibrous tumor should be included in the differential diagnosis of pediatric orbital tumors. PMID- 11020429 TI - Orbital compartment syndrome caused by intraorbital bacitracin ointment after endoscopic sinus surgery. AB - PURPOSE: To present an unusual case of orbital compartment syndrome after endoscopic sinus surgery. METHODS: Case report. RESULTS: Acute proptosis, chemosis, decreased vision, and ophthalmoplegia were found immediately after endoscopic sinus surgery. Ophthalmologic evaluation showed a tense orbit, and intraocular pressure increased to 54 mm Hg. Treatment was initiated and the intraocular pressure dropped. Computed tomography (CT) revealed the presence of bacitracin ointment in the orbit. CONCLUSION: Ophthalmic complications after sinus surgery are well identified. Postoperative orbital compartment syndrome may be caused by retrobulbar hemorrhage, edema, air (emphysema), or foreign material. In this case, the findings were caused by inadvertent injection of bacitracin ointment into the orbit. PMID- 11020431 TI - What makes clinical research ethical? emanuel EJ, wendler D, grady C. * JAMA 2000;283:2701-2711 PMID- 11020430 TI - NIH, under pressure, boosts minority health research. Helmuth L.* science 2000;288:596-597 PMID- 11020433 TI - Radiotherapy for Graves' orbitopathy: randomised placebo-controlled study. Mourits MP,* van kempen-harteveld ML, garcia MBG, koppeschaar HPF, tick L, terwee CB. Lancet 2000;355:1505-1509 PMID- 11020432 TI - The perilous state of academic medicine. Pardes H.**** JAMA 2000;283:2427-2429. Sustaining change in medical education. Griner PF,**** danoff D. JAMA 2000;283:2429-2431. The quest for the therapeutic organization. Bulger RJ.**** JAMA 2000;283:2431-2433. The evolving academic health center. Strategies and priorities at emory university. Saxton JF, blake DA, fox JT, johns MME.**** JAMA 2000;283:2434-2436. The academic health center. Some policy AB - + PMID- 11020434 TI - Treatment of ocular myasthenia with corticosteroids. Yes. Agius MA. ** arch neurol 2000;57:750-751. Treatment Of ocular myasthenia. Steroids only when compelled. Kaminski HJ,** daroff RB. Arch neurol 2000;57:752-753 PMID- 11020435 TI - The DNA sequence of human chromosome 21. Hattori M, fujiyama A, taylor TD, watanabe H, yada T, park HS, toyoda A, ishii K, totoki Y, choi DK, soeda E, ohki M, takagi T, sakaki Y,* taudien S, blechschmidt K, polley A, menzel U, delabar J, kumpf K, lehmann R, patterson D, reichwald K, rump A, shillhabel M, schudy A, zimmermann W, rosenthal A, kudoh J, shibuya K, kawasaki K, asakawa S, shintani A, sasaki T, nagamine K, mitsuyama S, antonakaris SE, minoshima S, AB - + PMID- 11020436 TI - Genetic and environmental factors in age-related nuclear cataracts in monozygotic and dizygotic twins. Hammond CJ,* sneider H, spector TD, gilbert CE. N engl J med 2000;342:1786-90 PMID- 11020438 TI - Nontraumatic corneal perforation. Lekskul M, fracht HU, cohen EJ,* rapuano CJ, laibson PR. Cornea 2000;19:313-319 PMID- 11020437 TI - The resiliency of the corneal endothelium to refractive and intraocular surgery. Edelhauser HF.* cornea 2000;19:263-273 PMID- 11020439 TI - Visual attention impairments in Alzheimer's disease. Rizzo M,* anderson SW, dawson J, myers R, ball K. Neurology 2000;54:1954-1959 PMID- 11020440 TI - Flow cytometric analysis of inflammatory markers in conjunctival epithelial cells in patients with dry eyes. Brignole F, pisella P-J, goldschild M, De saint jean M, goguel A, baudouin C.* invest ophthalmol vis sci 2000;41:1356-1363 PMID- 11020441 TI - Conjunctiva-associated lymphoid tissue in the human eye. Knop N, knop E.* invest ophthalmol vis sci 2000; 42:1270-1279 PMID- 11020442 TI - Use of in vitro methaemoglobin generation to study antioxidant status in the diabetic erythrocyte. AB - Poor glycaemic control in diabetes and a combination of oxidative, metabolic, and carbonyl stresses are thought to lead to widespread non-enzymatic glycation and eventually to diabetic complications. Diabetic tissues can suffer both restriction in their supply of reducing power and excessive demand for reducing power. This contributes to compromised antioxidant status, particularly in the essential glutathione maintenance system. To study and ultimately correct deficiencies in diabetic glutathione maintenance, an experimental model would be desirable, which would provide in vitro a rapid, convenient, and dynamic reflection of the performance of diabetic GSH antioxidant capacity compared with that of non-diabetics. Xenobiotic-mediated in vitro methaemoglobin formation in erythrocytes drawn from diabetic volunteers is significantly lower than that in erythrocytes of non-diabetics. Aromatic hydroxylamine-mediated methaemoglobin formation is GSH-dependent and is indicative of the ability of an erythrocyte to maintain GSH levels during rapid thiol consumption. Although nitrite forms methaemoglobin through a complex GSH-independent pathway, it also reveals deficiencies in diabetic detoxification and antioxidant performance compared with non-diabetics. Together with efficient glycaemic monitoring, future therapy of diabetes may include trials of different antiglycation agents and antioxidant combinations. Equalization in vitro of diabetic methaemoglobin generation with that of age/sex-matched non-diabetic subjects might provide an early indication of diabetic antioxidant status improvement in these studies. PMID- 11020443 TI - Pharmacology of the receptors for the phorbol ester tumor promoters: multiple receptors with different biochemical properties. AB - The phorbol ester tumor promoters and related analogs are widely used as potent activators of protein kinase C (PKC). The phorbol esters mimic the action of the lipid second messenger diacylglycerol (DAG). The aim of this commentary is to highlight a series of important and controversial concepts in the pharmacology and regulation of phorbol ester receptors. First, phorbol ester analogs have marked differences in their biological properties. This may be related to a differential regulation of PKC isozymes by distinct analogs. Moreover, it seems that marked differences exist in the ligand recognition properties of the C1 domains, the phorbol ester/DAG binding sites in PKC isozymes. Second, an emerging theme that we discuss here is that phorbol esters also target receptors unrelated to PKC isozymes, a concept that has been largely ignored. These novel receptors lacking kinase activity include chimaerins (a family of Rac-GTPase-activating proteins), RasGRP (a Ras exchange factor), and Unc-13/Munc-13 (a family of proteins involved in exocytosis). Unlike the classical and novel PKCs, these "non kinase" phorbol ester receptors possess a single copy of the C1 domain. Interestingly, each receptor class has unique pharmacological properties and biochemical regulation. Lastly, it is well established that phorbol esters and related analogs can translocate each receptor to different intracellular compartments. The differential pharmacological properties of the phorbol ester receptors can be exploited to generate specific agonists and antagonists that will be helpful tools to dissect their cellular function. PMID- 11020444 TI - Inhibition of cellular action of thrombin by N3-cyclopropyl-7-[[4-(1 methylethyl)phenyl]methyl]-7H-pyrrolo[3, 2-f]quinazoline-1,3-diamine (SCH 79797), a nonpeptide thrombin receptor antagonist. AB - A growing body of evidence suggests an important contribution of the cellular actions of thrombin to thrombosis and restenosis following angioplasty. Recently we reported on SCH 79797 (N3-cyclopropyl-7-?[4-(1-methylethyl)phenyl]methyl?-7H pyrrolo[3, 2-f]quinazoline-1,3-diamine) and its analogs as new potent, nonpeptide thrombin receptor antagonists. This study further characterizes the biochemical and pharmacological actions of pyrroloquinazoline inhibitors of protease activated receptor-1 (PAR-1) in human platelets and coronary artery smooth muscle cells (hCASMC). SCH 79797 and its N-methyl analog (SCH 203099) inhibited binding of a high-affinity thrombin receptor-activating peptide ([(3)H]haTRAP, Ala-Phe(p F)-Arg-ChA-HArg-[(3)H]Tyr-NH(2)) to PAR-1 with IC(50) values of 70 and 45 nM, respectively. SCH 79797 inhibited [(3)H]haTRAP binding in a competitive manner. SCH 79797 and SCH 203099 inhibited alpha-thrombin- and haTRAP-induced aggregation of human platelets, but did not inhibit human platelet aggregation induced by the tethered ligand agonist for protease-activated receptor-4 (PAR-4), gamma thrombin, ADP, or collagen. SCH 203099 inhibited surface expression of P-selectin induced by haTRAP and thrombin, and it did not increase P-selectin expression or prevent thrombin cleavage of the receptor. Thrombin and TFLLRNPNDK-NH(2) (TK), a PAR-1-selective agonist, produced transient increases in cytosolic free Ca(2+) concentration ([Ca(2+)](i)) in hCASMC. This increase in [Ca(2+)](i) was inhibited effectively by SCH 79797. However, the Ca(2+) transients induced by SLIGKV-NH(2,) a PAR-2-selective agonist, were not inhibited by SCH 79797. Thrombin- and TK stimulated [(3)H]thymidine incorporation also was inhibited completely by SCH 79797. The results of this study demonstrate that SCH 79797 and SCH 203099 are potent, selective antagonists of PAR-1 in human platelets and hCASMC. These data also suggest that the thrombin stimulation of Ca(2+) transients and mitogenesis in hCASMC is mediated primarily through activation of PAR-1. PMID- 11020445 TI - Daunorubicin cardiotoxicity: evidence for the importance of the quinone moiety in a free-radical-independent mechanism. AB - Anthracyclines, such as daunorubicin (Daun), and other quinone-containing compounds can stimulate the formation of toxic free radicals. The present study tests the hypothesis that the quinone moiety of Daun, by increasing free-radical production, disrupts sarcoplasmic reticulum (SR) function and thereby inhibits myocardial contractility in vitro. We compared Daun with its quinone-deficient analogue, 5-iminodaunorubicin (5-ID), using experimental interventions to produce various contractile states that depend on SR function. At concentrations of Daun or 5-ID that did not alter contractility (dF/dt) of steady-state contractions (1 Hz) in electrically paced atria isolated from adult rabbits, only Daun significantly attenuated the positive inotropic effects on dF/dt of increased rest intervals (PRP; post-rest potentiation) or increased stimulation frequencies. Attenuation was to 98+/-6% at 1 Hz, and 73+/-8 and 67+/-8% for 30 and 60 sec PRP, respectively, and 73+/-3 and 63 +/-3% at 2 and 3 Hz, respectively, for 88 microM Daun (P<0.05, vs pre-drug baseline values, mean +/- SEM). These effects of Daun were similar to those of caffeine (2 mM), an agent well known to deplete cardiac SR calcium. We also examined the effect of Daun in isolated neonatal rabbit atria, which lack mature, functional SR; Daun did not alter the force-frequency relationship or PRP contractions. Additional studies in Ca(2+)-loaded SR microsomes indicated that both Daun and 5-ID opened Ca(2+) release channels, with Daun being 20-fold more potent than 5-ID in this respect. Neither anthracycline, however, induced free-radical formation in SR preparations (assayed via nicking of supercoiled DNA) prior to stimulating Ca(2+) release. Thus, our results indicate that Daun impairs myocardial contractility in vitro by selectively interfering with SR function; the quinone moiety of Daun appears to mediate this cardiotoxic effect, acting through a mechanism that does not involve free radicals. PMID- 11020446 TI - Potentiation of 1-beta-D-arabinofuranosylcytosine-mediated mitochondrial damage and apoptosis in human leukemia cells (U937) overexpressing bcl-2 by the kinase inhibitor 7-hydroxystaurosporine (UCN-01). AB - Antileukemic interactions between the nucleoside analog 1-beta-D arabinofuranosylcytosine (ara-C) and the kinase inhibitor 7-hydroxystaurosporine (UCN-01) have been examined in relation to Bcl-2 expression/phosphorylation, mitochondrial damage, caspase activation, and loss of clonogenic potential. Subsequent exposure of ara-C-pretreated U937 cells (1 microM; 6 hr) to UCN-01 (300 nM; 24 hr) resulted in marked potentiation of pro-caspase-3 and -9 cleavage/activation, poly(ADP-ribose)polymerase degradation, diminished mitochondrial membrane potential (Deltapsi(m)), enhanced cytochrome c release, reduction in the S-phase fraction, and induction of classic apoptotic morphologic features. Enforced expression of full-length Bcl-2 significantly protected cells (at 24 hr) from ara-C/UCN-01-induced caspase activation and apoptosis, but was ineffective in preventing loss of Deltapsi(m) and cytochrome c release. Ectopic expression of a Bcl-2 N-terminal phosphorylation loop-deleted protein (Bcl 2Delta(32-80)) was more potent than its full-length counterpart in blocking drug induced loss of Deltapsi(m, ) caspase activation, and apoptotic morphology, but not cytochrome c release. Examination of cells at later intervals revealed that ectopic expression of Bcl-2 or Bcl-2Delta(32-80) could only delay, but not prevent, mitochondrial damage, caspase activation, and cell death induced by ara C/UCN-01 treatment. Despite their initial ability to inhibit apoptosis, neither full-length nor truncated Bcl-2 protein restored clonogenic potential to drug treated cells. These findings indicate that subsequent exposure of ara-C pretreated human leukemia cells to UCN-01 potently triggers mitochondrial damage and apoptosis, and that these events are postponed but not prevented by ectopic expression of Bcl-2 or its phosphorylation loop-deleted counterpart. PMID- 11020447 TI - Possible role of protein kinase C zeta in muscarinic receptor-induced proliferation of astrocytoma cells. AB - Recent studies have shown that protein kinase C zeta (PKC zeta) is part of a pathway that plays a key role in a wide range of physiological processes including mitogenesis, cell survival, and transcriptional regulation. Most studies on PKC zeta have been done by stimulating cells with tyrosine kinase receptor agonists, or by transfecting the cells with either constitutively active PKC zeta or negative mutants of PKC zeta. Less is known about the ability of endogenous G-protein-coupled receptors to generate a mitogenic signal through activation of endogenous PKC zeta. In the present paper, we showed that in 123 1N1 human astrocytoma cells, which express the G-protein-coupled M2, M3, and M5 muscarinic receptors, PKC zeta is activated by carbachol in a concentration dependent manner, resulting in the translocation of PKC zeta from the cytoplasm to granules in the perinuclear region. The effect of carbachol was long-lasting (up to 24 hr) and appeared to be mediated by activation of M3 muscarinic receptors. A selective PKC zeta inhibitor peptide (peptide Z) inhibited PKC zeta translocation as well as carbachol-induced DNA synthesis. Inhibition of both phosphatidylinositol 3-kinase and phospholipase D decreased carbachol-induced [(3)H]thymidine incorporation and blocked carbachol-induced PKC zeta translocation, suggesting an involvement of both pathways in these effects. PMID- 11020448 TI - In vitro effects of Se-allylselenocysteine and Se-propylselenocysteine on cell growth, DNA integrity, and apoptosis. AB - Two previously unevaluated selenium compounds, Se-allylselenocysteine (ASC) and Se-propylselenocysteine (PSC), have been shown recently to be active in the chemoprevention of experimentally induced mammary carcinogenesis. Other than their potential as chemopreventive agents, little is known about the pharmacological properties of these compounds. In this article, we report on the in vitro effects of ASC and PSC on cell growth inhibition, apoptosis, and the induction of DNA damage. The effects of ASC and PSC were examined in two mouse mammary epithelial cell lines derived from mammary hyperplasias. These cell lines, designated TM2H and TM12, have mutant or wild-type p53, respectively. It was observed that ASC but not PSC reduced, in a concentration- and time-dependent manner, the number of adherent cells in culture, and this suppressive effect was more prominent in TM12 than in TM2H cells. ASC was also found to induce alkaline labile DNA damage and the oxidation of pyrimidines, and it also increased the rate of apoptosis. These changes were not seen by exposure to PSC or the sulfur analog of ASC. However, additional data obtained from the intact rat mammary gland suggest that the loss of DNA integrity induced by ASC might not be manifest in vivo at doses of ASC that inhibit carcinogenesis. PMID- 11020449 TI - Inhibition by diallyl trisulfide, a garlic component, of intracellular Ca(2+) mobilization without affecting inositol-1,4, 5-trisphosphate (IP(3)) formation in activated platelets. AB - Garlic has been used in herbal medicine for thousands of years. Some reports have shown that garlic has protective effects against atherosclerosis and inhibits platelet function. In this study, we investigated the mechanism by which diallyl trisulfide (DT), a component of garlic, inhibits platelet function. DT inhibited platelet aggregation and Ca(2+) mobilization in a concentration-dependent manner without increasing intracellular cyclic AMP and cyclic GMP. DT also had no inhibitory effects on thromboxane A(2) (TXA(2)) production in cell-free systems. Collagen-related peptide (CRP)-induced Ca(2+) mobilization is regulated by phospholipase C-gamma2 (PLC-gamma2) activation. We evaluated the effect of DT on tyrosine phosphorylation of PLC-gamma2 and the production of inositol-1,4,5 trisphosphate (IP(3)). DT at concentrations that inhibited platelet aggregation and Ca(2+) mobilization had no effects on tyrosine phosphorylation of PLC-gamma2 or on the formation of IP(3) induced by CRP. Similar results were obtained with thrombin-induced platelet activation. DT inhibited platelet aggregation and Ca(2+) mobilization induced by thrombin without affecting the production of IP(3.) We then evaluated the effect of DT on the binding of IP(3) to its receptor. DT at high concentrations partially blocked the binding of IP(3) to its receptor. Taken together, our findings suggest that the agent suppresses Ca(2+) mobilization at a step distal to IP(3) formation. DT may provide a good tool for investigating Ca(2+) mobilization. PMID- 11020450 TI - Multidrug resistance protein functionality: no effect of intracellular or extracellular pH changes. AB - A major problem in the treatment of cancer is cellular resistance to cytotoxic drugs. In tumor cells in vitro, the development of multidrug resistance is usually accompanied by increased expression of drug transporters, either P glycoprotein (P-gp) or multidrug resistance-associated protein (MRP(1)). Both proteins belong to the superfamily of ATP-binding cassette (ABC) transporter proteins and mediate the transport of a broad range of drugs. Altenberg et al. (Proc Natl Acad Sci USA90: 9735-9738, 1993) have shown that changes in intra- or extracellular pH do not mediate P-gp-dependent multidrug resistance. Therefore, we similarly studied whether changes in intra- or extracellular pH could mediate MRP(1)-dependent multidrug resistance. In particular, we measured the MRP(1) mediated efflux of hydroxyrubicin from GLC4/ADR cells. Since hydroxyrubicin is a fully neutral anthracycline derivative that has no deprotonable function at pH lower than 10 and so cannot accumulate in non-nuclear compartments under the influence of pH or transmembrane gradients, we hypothesized that any modifications of its kinetics of efflux as a function of pH can be assigned to a modification of the transporter efficiency. However, as our data show, modifications of extra- and/or intracellular pH yielded no modification of the MRP(1)-mediated efflux of hydroxyrubicin. PMID- 11020451 TI - Effect of ethanol on the expression of hepatic glutathione S-transferase: an in vivo/in vitro study. AB - Ethanol, a human toxicant and a solvent in pharmacological research, is known to interfere with biotransformation of xenobiotics. We compared the in vivo and in vitro long-term effects of ethanol exposure on the expression of glutathione S transferases (GST, EC 2. 5.1.18) in rat liver. Long-term in vivo ethanol treatment to achieve blood ethanol levels ranging between 10-50 mM was by liquid diet feeding. For in vitro experiments, rat hepatocytes co-cultured with rat liver epithelial cells were exposed to 17 and 68 mM ethanol for up to 10 days. Two weeks of liquid diet ethanol treatment increased total GST activity. Both Mu and Alpha classes and in particular the A1 and A2 subunits and the amount of their corresponding mRNAs were increased. Total GST activity was also increased in co-cultures after exposure to 68 mM ethanol for 10 days. However, the Mu class subunits M1 and M2 and the corresponding mRNAs were increased, rather than the Alpha class subunits. Thus, long-term exposure to ethanol induces hepatic GST both in vivo and in vitro, but different isoenzymes are affected. Consequently, extrapolation of in vitro data on GST expression and regulation to the in vivo situation must be judicious. During xenobiotic metabolism in cell culture, a shift in relative expression and induction of different GST forms may occur, resulting in either an under- or overestimation of effects. PMID- 11020452 TI - Characterisation of the antitrypanosomal activity of peptidyl alpha-aminoalkyl phosphonate diphenyl esters. AB - Two groups of irreversible serine peptidase inhibitors, peptidyl chloromethyl ketones and peptidyl phosphonate diphenyl esters, were examined for antitrypanosomal activity against the bloodstream form of Trypanosoma brucei brucei. Both peptidyl chloromethyl ketones and peptidyl phosphonate diphenyl esters inhibited trypsin-like peptidases of the parasites and exhibited antitrypanosomal activity at micromolar concentrations. In live T. b. brucei, labelled analogues of both of these groups of inhibitors primarily targeted an 80 kDa peptidase, possibly a serine oligopeptidase known as oligopeptidase B. In an in vivo mouse model of infection, one of these inhibitors, carbobenzyloxyglycyl-4 amidinophenylglycine phosphonate diphenyl ester, was curative at 5 mg kg(-1) day( 1) but appeared toxic at higher doses. There was no significant correlation between the inhibitory potency (as evaluated against purified T. b. brucei oligopeptidase B) and the in vitro antitrypanosomal efficacy of either group of inhibitors, suggesting that these inhibitors were acting on multiple targets within the parasites, or had different cell permeability properties. These findings suggest that serine peptidases may represent novel chemotherapeutic targets in African trypanosomes. PMID- 11020453 TI - Phosphorylation of the anti-HIV compound (S,S)-isodideoxyadenosine by human recombinant deoxycytidine kinase. AB - (S,S)-Isodideoxyadenosine [(S,S)-isoddA] is an anti-HIV active compound discovered in our laboratory. However, its cellular mechanism of action, particularly the critical first stage of phosphorylation, is not understood. IsoddA is not phosphorylated by adenosine kinase. Also, because it is not a substrate for adenosine deaminase, it would not be activated by the pathway taken by ddA, i. e. via 5'-nucleotidase phosphorylation of ddI and conversion of ddIMP to ddAMP. However, we have discovered that human recombinant 2'-deoxycytidine kinase (dCK) phosphorylates (S,S)-isoddA. The enzyme kinetic data revealed that the extent of monophosphorylation of this L-related nucleoside was comparable to that found with ddA. (S,S)-IsoddATP is among the most potent inhibitors of HIV reverse transcriptase known, which suggests that the observed low efficiency of phosphorylation of this compound by dCK is a key factor that limits the capacity of human lymphocytes to make (S,S)-isoddA an exceptionally active anti-HIV agent. PMID- 11020454 TI - Selective induction of cytochrome P450 3A1 by dexamethasone in cultured rat hepatocytes: analysis with a novel reverse transcriptase-polymerase chain reaction assay section sign. AB - The study of drug metabolism in cultured rat hepatocytes is hampered by the rapid loss of the expression of cytochrome P450 enzymes. Nevertheless, the activity of cytochrome P450 3A (CYP3A), one of the most important isoenzymes for drug metabolism, can be elevated by chemical inducers. In the present study, we investigated in cultured rat hepatocytes the induction of all four currently identified CYP3A isoforms by dexamethasone, and compared the results obtained in vitro with the induction profile of dexamethasone in vivo. To this end, CYP3A mRNA levels were quantified with a novel, radioactive reverse transcriptase polymerase chain reaction (RT-PCR) assay, and CYP3A enzymatic activity was measured by a testosterone hydroxylation assay. In the RT-PCR assay, CYP3A isoforms were co-amplified with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the presence of radioactively labeled nucleotides. This resulted in an extremely sensitive and accurate determination of CYP3A expression levels, relative to those of GAPDH. Using this RT-PCR assay, it was found that the expression of all CYP3A isoforms in rat hepatocytes, cultured on a collagen matrix, was decreased by 80-90% within one day of cultivation. After addition of dexamethasone, at one day after isolation, CYP3A1 mRNA levels were elevated to levels comparable to those in freshly isolated hepatocytes within two days. In contrast, CYP3A2, CYP3A9, and CYP3A18 mRNA levels were not affected by dexamethasone treatment, and were hardly detectable after three days of cultivation. CYP3A enzymatic activity was also induced in cultured hepatocytes (approximately 6-fold) after addition of dexamethasone. In vivo, CYP3A1 mRNA levels increased 45-fold after dexamethasone administration. However, in contrast to the situation in cultured hepatocytes, CYP3A2 and CYP3A18 were also induced, albeit to a lesser extent (4- and 7-fold elevated mRNA levels, respectively). We conclude that the selective induction of CYP3A1 in dexamethasone-treated rat hepatocytes allows the study of biotransformation reactions by CYP3A1, without interference by any of the other CYP3A isoenzymes. PMID- 11020455 TI - Obstacles to the prediction of estrogenicity from chemical structure: assay mediated metabolic transformation and the apparent promiscuous nature of the estrogen receptor. AB - Information on structure-activity relationships (SAR) and pathways of metabolic activation would facilitate the preliminary screening of chemicals for estrogenic potential. Published crystallographic studies of the estrogen receptor (ER) imply an essential role of the two hydroxyl groups on estradiol (17beta-E(2)) for its binding to ER. The influence of these hydroxyl groups on ER binding and estrogenicity was evaluated by the study of 17beta-E(2) with one or both of these hydroxyl groups removed (17beta-desoxyestradiol and 3, 17beta-bisdesoxyestradiol, respectively). 6-Hydroxytetralin (17beta-E(2) with its C- and D-rings removed) and other synthetic estrogens were also studied. The estrogenicity assays comprised a yeast ER-mediated transcription assay, mammalian cell transcription assays incorporating either ER alpha or ER beta, and the immature rat uterotrophic assay. With the exception of 6-hydroxytetralin in the uterotrophic assay, all the chemicals were active in all the assays. Hydroxylation of the two desoxy compounds to estradiol was shown to occur in immature female rats, but metabolism was not implicated in the responses observed in the ER-binding and yeast systems. It is concluded that the 3-hydroxyl and 17beta-hydroxyl groups of 17beta-E(2) are not absolute requirements for estrogenicity. It would therefore be of value to the derivation of SAR for estrogenicity were the crystal structure of the bisdesoxy-E(2)/ER complex to be evaluated. PMID- 11020456 TI - Stimulation of hepatocyte growth factor production in human fibroblasts by the protein phosphatase inhibitor okadaic acid. AB - In this study, we examined whether the production of hepatocyte growth factor (HGF) in fibroblasts is regulated by protein phosphatase(s). Inhibitors of the enzymes okadaic acid and calyculin A were used for this purpose. Both inhibitors markedly stimulated HGF production in human skin fibroblasts in a dose-dependent manner. The effects of okadaic acid and calyculin A were maximal at 25-37.5 and 1.25 nM, respectively. Highly active HGF production in MRC-5 human embryonic lung fibroblasts was also promoted by both inhibitors. The effect of okadaic acid was accompanied by an up-regulation of HGF gene expression. The stimulating effect of okadaic acid on HGF production was synergistic with that of phorbol 12-myristate 13-acetate (PMA) and epidermal growth factor (EGF), whereas it was additive to the effect of cholera toxin. The protein kinase C (PKC) inhibitor GF 109203X inhibited the effect of PMA, but not of okadaic acid and EGF. The effect of okadaic acid as well as EGF was not inhibited, but rather enhanced in human skin fibroblasts pretreated for 24 hr with a high dose of PMA to deplete PKC, as compared with its effect in untreated cells. PD 98059, an inhibitor of mitogen activated protein (MAP) kinase kinase, suppressed the effects of okadaic acid and EGF, but not those of cholera toxin and 8-bromo-adenosine 3',5'-cyclic monophosphate (cAMP). These results suggest that HGF production in human skin fibroblasts is down-regulated by protein phosphatase(s) and that HGF production stimulated by okadaic acid is, at least in part, dependent on the activation of the MAP kinase cascade. PMID- 11020458 TI - The role of automated urine particle flow cytometry in clinical practice. AB - Urine particle flow cytometers (UFC) have improved count precision and accuracy compared to visual microscopy and offer significant labor saving. The absence of an internationally recognized reference measurement procedure, however, is a serious drawback to their validation. Chamber counting by phase contrast microscopy of supravitally-stained uncentrifuged urine is considered the best candidate for reference. The UF-100 (Sysmex Corporation, Japan) identifies RBC, WBC, squamous epithelial cells, transitional epithelial and renal tubular cells (SRC), bacteria, hyaline and inclusional casts, yeast-like cells, crystals and spermatozoa, using argon laser flow cytometry. Evaluations have established acceptable linearity over useful working ranges, with an imprecision that is consistently and significantly less than microscopy, and with negligible carry over. Comparisons of UFC with chamber counts, quantitative urine microscopy, sediment counts, test strips, bacterial culture and urine density are reviewed. Clinical studies include diagnosis and monitoring of urinary tract infection; localization of the sites of hematuria; and diagnosis, monitoring and exclusion of renal disease. The most popular approach is to combine test strips with UFC for primary screening either always by both methods or by using test strips for analytes unrelated to particles analyzed by UFC. Expert systems now exist combining both test modalities based on user definable decision rules. The implementation of such a strategy significantly reduces microscopy review and saves time and expense without diminishing clinical utility. PMID- 11020457 TI - Synergy between ethanol and grape polyphenols, quercetin, and resveratrol, in the inhibition of the inducible nitric oxide synthase pathway. AB - In atherosclerosis and tumor initiation, inducible nitric oxide synthase (iNOS) has been implicated in the damage of vascular walls and DNA, respectively. Moderate consumption of red wine has been ascribed as a preventive for coronary heart disease; however, there has been much debate over whether the beneficial effect is from grape polyphenolic components or ethanol. We studied the interaction of grape compounds on nitric oxide (NO) production by macrophages, mediators of blood vessel damage in atherosclerosis. For the murine macrophage cell line RAW 264.7, stimulation with lipopolysaccharide and interferon-gamma led to expression of the iNOS gene and production of NO. The polyphenols quercetin and resveratrol at a micromolar range suppressed iNOS gene expression and NO production, as determined by reverse transcription-polymerase chain reaction and nitrite assay. The polyphenols were also found to be scavengers of NO in an acellular system using sodium nitroprusside under physiological conditions. Ethanol, at a moderate level, did not produce any appreciable level of reduction of iNOS or NO activity. However, its presence at 0.1 to 0.75% enhanced the effect of grape polyphenols concentration-dependently. Thus, the interaction between these components plays a significant role in the health effects of red wine, especially with respect to their effect on the NO pathway. PMID- 11020460 TI - Rapid serum vancomycin assay by high-performance liquid chromatography using a semipermeable surface packing material column. AB - A new isocratic high-performance liquid chromatographic (HPLC) assay has been developed for vancomycin that uses direct injection of microquantities of serum into a separation column filled with octyl-C(8) silica support that has a semipermeable surface. A mixture of disodium hydrogen phosphate buffer (pH 7.0) and acetonitrile is used as the mobile phase, and vancomycin is directly detected at 240 nm. The minimum limit of detection was 0.5 microg/ml at a signal-to-noise ratio of 3:1. Linearity was established from 0 to 100 microg/ml. The coefficient of variation for within-run reproducibility was 1.1-2.7% for a concentration range of 2.9-52.5 microg/ml; for day-to-day reproducibility it was 4.0% and 3.1% for a concentration range of 5.8-26.4 microg/ml, and the recovery rate was 94 105%. There was no interference from 41 antibiotics or other drugs currently in use. The correlation coefficient between the fluorescence polarization immunoassay (x) and this method (y) was 0.995 with a linear equation, y = 1.06x - 0.924. This method is simple, rapid, and provides an economical quantification of serum vancomycin. PMID- 11020459 TI - Plasma concentration of brain natriuretic peptide as a biochemical marker for the evaluation of right ventricular overload and mortality in chronic respiratory disease. AB - The purpose of this study was to evaluate whether the plasma brain natriuretic peptide (BNP) concentration is a useful marker of right ventricular (RV) overload and whether it has prognostic value as a predictor of death in patients with chronic respiratory disease (CRD). We measured the plasma BNP and atrial natriuretic peptide (ANP) concentrations in 31 consecutive patients with CRD who underwent right-heart catheterization to evaluate pulmonary hypertension. All patients were followed for >12 months. The plasma BNP concentration closely correlated with the mean pulmonary artery pressure and pulmonary vascular resistance (r=0.62, P<0.0005 and r=0. 85, P<0.0001), and showed a weak linear correlation with cardiac output (r=-0.36, P<0.05). During the follow-up period, 5 (16%) end-stage CRD deaths (4 RV heart failure and 1 respiratory infection) and 2 non-end-stage CRD deaths occurred. In a stepwise multivariate Cox proportional hazards regression analysis including age, sex, BNP, ANP, hemodynamic variables and the ratio of PaO(2) to fraction of inspired oxygen, only BNP (P<0.05) was an independent predictor of end-stage CRD death. The upward and leftward shift in the receiver operating characteristic curve between patients with end-stage CRD death and those without was greater for BNP than for ANP. Our findings suggest that the plasma BNP concentration may be an inexpensive, simple and useful marker of RV overload and end-stage CRD death in CRD patients. These preliminary results need to be confirmed in a large series of CRD patients. PMID- 11020461 TI - Comparative antioxidant potential of anaesthetics and perioperative drugs in vitro. AB - We have investigated the comparative antioxidant capacity of a range of anaesthetics (inhaled and intravenous) and perioperative neurosurgical drugs (at clinically relevant concentrations) using different radical species and assay methods in vitro. The highest levels of antioxidant activity against the ABTS(.+) radical were obtained with propofol (100 mmol/LTE) and dopamine (1080 mmol/LTE), respectively. However, only dopamine (12 mmol/l) showed antioxidant activity in protecting proteins in normal brain tissue from oxidative damage (assessed via SDS-PAGE analysis) induced by OH(.) or O(2)(-.) generated radiolytically in vitro. Neither dopamine nor propofol showed antioxidant activity against O(2)(-.) generated chemically via reaction between xanthine and xanthine oxidase in vitro. From these data, together with data on the relative antioxidant properties of anaesthetics/drugs obtained by other research groups which we have reviewed, we conclude that the apparent antioxidant activity of a given compound may depend entirely on the free radical species and/or the method of generation or assay employed. Finally, we suggest that on the basis of data obtained showing protection of brain proteins from oxidative damage induced by OH(.), or O(2)(-.) in vitro, further investigation into the in vivo antioxidant therapeutic potential of dopamine (or its analogues) on neurosurgical patients may be warranted. PMID- 11020462 TI - Marked elevation in serum apolipoprotein E in a case of heterozygous cholesteryl ester transfer protein deficiency. AB - The subject was a 57-year-old Japanese woman with a body mass index of 21.2 kgm( 2). Her serum total cholesterol (TC), triglycerides (TG) and HDL-cholesterol levels were 7.11 mmoll(-1), 0.53 mmoll(-1) and 2.05 mmoll(-1), respectively. She had a marked increase of serum apolipoprotein (Apo) E concentration of 25 mgdl( 1) with normal concentrations of serum Apo A-I, A-II, B, C-II and C-III. Polymerase chain reaction-restriction fragments length polymorphism analysis of the cholesteryl ester transfer protein (CETP) gene from this subject revealed the heterozygous nucleotide change causing a Asp442 to Gly substitution (D442G) in the CETP protein. For comparison, 11 unrelated female subjects with this mutation (age, 57+/-5.1 years; BMI, 22+/-1.5 kgm(-2); TC, 7.23+/-1.16 mmoll(-1); TG, 1.44+/-0.80 mmoll(-1); HDL-C, 2.47+/-0.53 mmoll(-1)) were found to have a serum Apo E concentration of 7+/-1.5 mgdl(-1), about a third of the patient's concentration. The lipoprotein profile of the proband's serum analyzed by disk polyacrylamide gel electrophoresis showed a trace amount of VLDL. A vitamin A fat loading test showed little increase in serum triglycerides and retinyl palmitate levels compared with control subjects at 2, 4 and 6 h after fat loading. Ultracentrifugation analysis of her serum revealed no detectable Apo E in the VLDL fraction but showed a large amount of Apo E in the HDL fraction, in contrast to a normal control, who had Apo E in the VLDL fraction as well as in the HDL fraction. Sequence analysis of the Apo E gene from the subject showed no nucleotide changes in exon 3 and exon 4, which code the mature Apo E protein, indicating there is no structural abnormality in the Apo E protein. Direct sequence analysis of the LDL receptor gene also did not show any nucleotide change. Based on these findings, it was hypothesized that the marked increase of Apo E in the patient's serum was caused by a decreased transfer of Apo E from HDL particles to TG-rich lipoproteins or impaired uptake of Apo E-containing HDL by LDL receptor or remnant receptor, due presumably to a dysfunction of these receptors in the patient. PMID- 11020463 TI - Evidence that pioglitazone, metformin and pentoxifylline are inhibitors of glycation. AB - Enhanced formation and accumulation of advanced glycation end products (AGEs) have been proposed to play a major role in the pathogenesis of diabetic complications, and atherosclerosis, leading to the development of a range of diabetic complications including nephropathy, retinopathy and neuropathy. Several potential drug candidates as AGE inhibitors have been reported recently. Aminoguanidine is the first drug extensively studied. However, there are no currently available medications known to block AGE formation. We have previously reported a number of novel and structurally diverse compounds as potent inhibitors of glycation and AGE formation. We have now studied several of the existing drugs, which are in therapeutic practice for lowering blood sugar or the treatment of peripheral vascular disease in diabetic patients, for possible inhibitory effects on glycation. We show that that three compounds; pioglitazone, metformin and pentoxifylline are also inhibitors of glycation. PMID- 11020464 TI - Are equally spaced specimen collections necessary to assess biological variation? Evidence from renal transplant recipients. AB - The established method for determining the components of biological variation (BV) requires equispaced time intervals between samplings. In a previous study, we determined BV in renal post transplantation patients, taking advantage of the samples obtained within their clinical treatment protocol (not necessarily equispaced). To confirm the validity of this practice, we sought to determine if the use of varying sampling intervals has an effect on the results obtained in such biological variation studies. The study included two phases: comparison of the results found with identical and non-identical sampling intervals and correlation between the within-subject BV and the length of the sampling interval. There were no differences in within-subject BV between the groups or correlations with sampling intervals for any of the constituents studied. We conclude that samples acquired within established clinical protocols for kidney transplant recipients can be used for estimating BV. PMID- 11020465 TI - Comparison of the determination of superoxide dismutase and antioxidant capacity in neurological patients using two different procedures. AB - As oxidative stress in relation with neurological diseases has become an important point in recent research, simple methods to be used in epidemiological studies and clinical practice are required. The hypothesis that the analytical methods used in research laboratories (RLM) can be used interchangeably with commercial kits (CKM) for SOD and TRAP is tested. Both methods were compared using linear transformations of the RLM measurements into the CKM scales. Data were obtained from Alzheimer's, Parkinson's, and vascular dementia patients and controls. The lack of fit and the run's test of residuals were not significant, but the same sign method detected significant nonlinearities (P<0.000001 for SOD, P<0.01 for TRAP). The intragroup CVs of both methods were comparable for TRAP, while in the RLM determinations of SOD resulted in <50% of those obtained with the CKM. The ANCOVA comparison of the regression parameters across the clinical groups resulted significant for SOD (P<0.0001) and not significant for TRAP. Both methods agree in describing the features of the clinical groups, but the degree of agreement at the individual concentration was poor and they could not be readily intercalibrated. Normal and pathological values should be obtained independently for the CKM to insure their applicability to large populations. PMID- 11020466 TI - Particle size analysis of high density lipoproteins in patients with genetic cholesteryl ester transfer protein deficiency. AB - We investigated the detailed distribution of high-density lipoproteins (HDL) particle size in patients with cholesteryl ester transfer protein (CETP) deficiency. Serum samples pre-stained with Sudan black B were electrophoresed using 4-30% polyacrylamide gradient gels, and the Stokes diameter of HDL particles was determined in 23 patients with genetic CETP deficiency, nine patients with hyperalphalipoproteinemia and seven subjects with normal HDL cholesterol concentrations. The mean Stokes diameter of HDL particles in CETP deficient patients (11.2+/-0.6 nm) was significantly greater than hyperalphalipoproteinemia (10.7+/-0.3 nm, P<0.05) and normal subjects (9.5+/-0.4 nm, P<0.01). A significant relationship was found between mean HDL size and serum CETP mass concentrations (P<0.05). When the particle size of all detected HDL bands was investigated, extra-large HDL particles larger than 12 nm were found in 14 of the 23 patients with CETP deficiency, which were not found in any of the hyperalphalipoproteinemia patients or normal subjects. Serum low-density lipoproteins (LDL) cholesterol and total cholesterol concentrations were lower in CETP deficiencies with extra-large HDL particles than those in non-carriers (P<0.01). These results indicate that extra-large HDL may be an index to clarify the relationship between genetic CETP deficiency and atherosclerosis. PMID- 11020467 TI - Diminished rate of mouse peritoneal macrophage cholesterol efflux is not related to the degree of HDL glycation in diabetes mellitus. AB - The efflux of (14)C-cholesterol from mouse peritoneal macrophages mediated by in vivo and in vitro glycation of intact HDL(3) and by HDL(3) apolipoproteins was investigated. Cholesterol-laden cells were incubated a long time with HDL(3) from control subjects (C), poorly controlled diabetes mellitus patients (D) and with HDL C submitted to in vitro glycation (G), as well as with all their respectively isolated apolipoproteins. A diminished cholesterol efflux rate occurred in incubations with intact HDL(3) D but not with intact HDL(3)G or with apoHDL(3)C, G or D. The specific binding of (125)I-HDL(3)G to the cell receptor, obtained upon incubation in the absence and in the presence of excess unlabelled HDL(3), was lower than the control. The role of apoE secretion by cholesterol-laden macrophages on cholesterol efflux was analyzed by incubating apoE knockout and control mice macrophages with HDL C or HDL G: a lower cholesterol efflux was observed from apoE knockout macrophages but glycation of HDL(3) did not influence this process either. The diminished capacity to remove cholesterol by the HDL drawn from diabetic subjects must be attributed to other modifications of the lipoproteins, except for non enzymatic glycation. Thus, events that impair the cell cholesterol removal in diabetes mellitus are multifaceted. PMID- 11020468 TI - Relationship between soluble thrombomodulin in plasma and coagulation or fibrinolysis in type 2 diabetes. AB - Serum concentration of soluble thrombomodulin (TM) is thought to be a marker for endothelial damage. Although several studies have reported that serum TM concentrations are increased in patients with diabetes mellitus, there is little information on the physiological function of soluble TM in human plasma. To evaluate the relationship of soluble TM in plasma between coagulation and/or fibrinolysis system in patients with diabetes, we measured plasma soluble TM, protein C activity (a natural anticoagulant induced by thrombin-TM complex), prothrombin F1+2 (a direct marker of thrombin generation), and plasmin-alpha 2 antiplasmin complex (PAP) and D dimer (measures of fibrinolytic activity) in 55 patients with type 2 diabetes mellitus. The plasma concentrations of soluble TM (P<0.01), protein C activity (P<0.01), prothrombin F1+2 (P<0.05), PAP (P<0.001) and D dimer (P<0.001) were significantly higher in the diabetic patients than the 48 age-matched control subjects. The plasma concentrations of TM and PAP were obviously increased in patients with diabetic nephropathy. In the diabetic patients, the plasma concentrations of soluble TM were inversely correlated with the protein C activity (r=-0.43, P<0.005), and were positively correlated with the plasma concentrations of prothrombin F1+2 (r=0.63, P<0.0001) and the plasma PAP concentrations (r=0.30, P<0.05). The present study demonstrated that both coagulation and fibrinolysis are enhanced concomitantly in patients with type 2 diabetes mellitus, and that an increase in plasma concentration of soluble TM is associated not only with hypercoagulability but also with enhanced fibrinolysis in diabetic patients. PMID- 11020469 TI - Electrophoretic study of tartrate-resistant acid phosphatase isoforms in endstage renal disease and rheumatoid arthritis. AB - The objective of this study was to identify the isoform, type-5a or type-5b, responsible for increased tartrate-resistant acid phosphatase (TRAP) activity in endstage renal disease (ESRD) and TRAP protein in rheumatoid arthritis (RA). We studied 24 sera each from healthy, ESRD and RA subjects. Type-5 TRAP activity and protein were quantitated by immunoassays. Isoform expression was determined by computerized imaging of non-denaturing polyacrylamide gels (PAGE) stained for TRAP activity. Other biochemical markers included: intact parathyroid hormone (iPTH), total and bone-specific alkaline phosphatase (TAP, BAP), N-telopeptides of type-I collagen (NTx), and free pyridinoline (Pyd). Isoform 5a was normal in both ESRD and RA. Isoform 5b was elevated in ESRD only. Serum TRAP activity correlated with both isoforms 5a and 5b in RA, but only with 5b in ESRD. TRAP protein assays did not correlate with PAGE assays for 5a or 5b. TRAP activity, but not protein, correlated with BAP and NTx in RA sera. Both TRAP activity and protein correlated with iPTH, TAP and Pyd in ESRD sera. Increased TRAP activity in ESRD was due to increased osteoclastic isoform 5b and related to bone turnover. Increased TRAP protein in RA was suspected, but not proven, to be isoform 5a and not related to bone turnover. Heterogeneity of serum TRAP and preferential expression of isoforms has clinical significance in different diseases including ESRD and RA. PMID- 11020470 TI - Hyaluronidase activity in serum of patients with monoclonal gammapathy. AB - Hyaluronidase and hyaluronic acid, two substances thought to be strongly implicated in carcinogenesis, were assessed in the plasma of 35 patients with newly documented monoclonal gammapathy and in 25 control patients. A significant increase was found in plasma hyaluronidase activity in the patients with monoclonal gammapathy. A statistically significant positive correlation was found between hyaluronidase activity and monoclonal immunoglobulin levels in plasma. An increase in serum hyaluronidase activities may be a response to the deleterious effect of hyaluronic acid in cell migration and tumor progression. Further studies are needed to assess the value of hyaluronidase activity as a marker of tumor progression. PMID- 11020471 TI - Influence of sex and age on serum nitrite/nitrate concentration in healthy subjects. AB - Measurement of serum nitrite/nitrate (NOx) concentrations has been considered useful to estimate nitric oxide production in humans. However little is known about the physiologic range and the factors affecting serum NOx levels. The aim of this study was, thus, to investigate the influence of sex and age on serum NOx levels in healthy subjects. We selected 263 healthy subjects (118 women and 145 men, 20-69 y) from 505 consecutive subjects who received annual medical checkups at our hospital. Serum NOx levels were determined using an analyzer employing the Griess method. The linear regression analysis showed that NOx increased significantly according to age in women (r=0.22, P<0.05), but did not in men (P=NS). Women of the younger age group (<40 y) showed significantly lower NOx levels than men of the same age group (P<0.05), whereas there was no significant difference between men and women of the older age (> or =40 y). Then, to investigate whether menopause affects serum NOx levels, middle aged women (46-55 y) were selected. Multiple logistic regression analysis showed that menopause was an independent factor for increased serum NOx levels in middle aged women (r=0.4, P<0.05). These results suggest that the serum NOx concentration is affected by age in healthy women, possibly depending on menopausal state. PMID- 11020472 TI - N alpha-methylhistamine: association with Helicobacter pylori infection in humans and effects on gastric acid secretion. AB - Infection with the bacterium Helicobacter pylori is associated with altered gastric acid secretion and gastrointestinal disease. Recent work has suggested that N alpha-methylhistamine, produced by the bacterium and acting on histamine receptors in gastric tissue, might be involved. Gastric juice and tissue biopsies from infected patients have been analysed for the presence of N alpha methylhistamine using a specific and sensitive assay based on gas chromatography mass spectrometry. N alpha-Methylhistamine was detected in five of seven samples of gastric juice from infected patients (5-180 pmol/ml) but was absent in nine uninfected subjects. The compound was not found in fundic and antral biopsies from both subject groups. Helicobacter pylori, cultured on agar and in broth with and without added histamine, was found not to produce detectable levels of N alpha-methylhistamine. Instillation of this compound at 10(-5) mol/l into the gastric lumen produced a significant increase in acid secretion in vivo while plasma gastrin concentration remained unchanged. N alpha-Methylhistamine in gastric juice appears therefore to be associated with infection, although this product is not generated directly by the bacterium. The concentrations found are below those required to affect acid secretion or gastrin production in vivo, although higher local concentrations may exist around a site of infection. PMID- 11020473 TI - An automated assay for measuring serum ascorbic acid with use of 4-hydroxy 2,2,6,6-tetramethylpiperidinyloxy, free radical and o-phenylenediamine. AB - We developed a novel, cost-effective, and automated assay for ascorbic acid (AsA) in serum using a COBAS MIRA S analyzer (Roche Diagnostic System). Our method has a wide dynamic range and covers AsA concentrations from well below the lower reference interval to well above it. AsA is oxidized by 4-hydroxy-2,2,6,6 tetramethylpiperidinyloxy, free radical (TEMPO) to dehydroascorbic acid (DAsA). The latter condenses with o-phenylenediamine (OPDA) to form a quinoxaline derivative that absorbs light at 340 nm. The change in absorbance at 340 nm is proportional to the concentration of AsA in the specimen. The automated system permitted the assay of 65 specimens per hour at a cost of approximately US$ 0.01 per specimen for reagents. The assay can be applied directly to serum specimens (direct method) and also to sera with a prior deproteinization step with metaphosphoric acid. The detection limit for the direct serum assays is 0.8 vs. 0.4 mg/l with the deproteinization method. The recovery of AsA from a supplemented serum pool was of >95% for both procedures. We used four distinct methods on 66 patients sera. The direct method for AsA correlated well with an HPLC method (r=0.964, P<0.001); the direct method also correlated well with a method that uses AsA oxidase (r=0.975, P<0. 001). The deproteinization method correlated well with HPLC (r=0.981, P<0.001), and with the AsA oxidase procedure (r=0.994, P<0.001). Ten within-day determinations on a serum pool gave a C.V. <4.3% for both the direct and deproteinization procedures. The between-day assays of the same serum pool over 10 days gave a C.V. of <6.7% by both methods. PMID- 11020474 TI - Single strand conformation polymorphism analysis of K-ras gene mutations by capillary electrophoresis with laser-induced fluorescence (LIF) detector. AB - Mutations of K-ras gene play an important role in neoplastic progression. The capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) technique is available for the detection of gene mutations. Using an automated capillary electrophoresis with short-chain linear polyacrylamide, after denaturation of PCR products, injections were performed at reverse polarity of 5 kV for 15 s and the separations were carried out under a constant voltage of 8 kV. Of 16 specimens of lung cancer tissue, two specimens were found to have abnormal peaks in the electrophoretogram. CE-SSCP is rapid, automated, and has high performance. PMID- 11020475 TI - Measurable amount of glutathione in blood is influenced by oxygen saturation of hemoglobin. PMID- 11020476 TI - Detection of methylenetetrahydrofolate reductase (MTHFR) C677T and prothrombin G20210A mutations: second restriction site for digestion control of PCR products. PMID- 11020477 TI - Erratum to "Interpreting complex urinary patterns with MDI LABLINK: a statistical evaluation". PMID- 11020478 TI - Antagonist binding at 5-HT(2A) and 5-HT(2C) receptors in the rabbit: high correlation with the profile for the human receptors. AB - This study examined the binding of serotonin receptor antagonists at the 5-HT(2A) and 5-HT(2C) receptors of the rabbit's cerebral cortex. The 5-HT(2A) receptor was characterized by the binding of [3H]MDL 100,907 (R(+)-alpha-(2, 3 dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine-methan ol) to cortical membranes and the 5-HT(2C) receptor by the binding of [3H]mesulergine in the presence of the selective 5-HT(2A) receptor ligand spiperone. Both [3H]MDL 100,907 and [3H]mesulergine demonstrated high affinity binding to single sites in rabbit membranes. Based on Scatchard plots of [3H]MDL 100,907 binding, the mean B(max) was 8.5+/-0.7 fmol/mg tissue and the mean K(d) was 33. 1+/-3.5 pM. For [3H]mesulergine binding the mean B(max) was 3.70+/-0. 58 fmol/mg tissue and the mean K(d) was 0.35+/-0.05 nM. Binding of [3H]MDL 100,907 to the 5-HT(2A) receptor and of [3H]mesulergine to the 5-HT(2C) receptor was confirmed by displacement studies with highly selective 5-HT(2A) and 5-HT(2C) receptor ligands. The pharmacological profile of these ligands in rabbits correlated highly with published values for 5-HT(2A) (r=0.91, P<0.001) and 5-HT(2C) (r=0.94, P<0.001) receptors in humans. There was also a high correlation between the profiles for human and rat 5-HT(2C) receptor (r=0.92, P<0.001), but not for 5-HT(2A) receptors (r=0.53, P>0.10). It was concluded that the rabbit provides an appropriate animal model for studies attempting to predict the pharmacology of human 5-HT(2A) and 5 HT(2C) receptors. PMID- 11020479 TI - A key role for the mitochondrial benzodiazepine receptor in cellular photosensitisation with delta-aminolaevulinic acid. AB - The aim of this study was to determine the part played by the mitochondrial benzodiazepine receptor in cellular photosensitisation with the protoporphyrin IX precursor, delta-aminolaevulinic acid. Evaluation of the delta-aminolaevulinic acid-concentration dependence and kinetics of fluorescent protoporphyrin IX accumulation in monolayers of rat AR4 2J pancreatoma cells established a basis for assessing pharmacological modulation of the biosynthetic pathways for protoporphyrin IX production and photocytotoxicity. Iron chelation enhanced the accumulation of photo-active protoporphyrin IX whereas 1-(2-chlorophenyl)-N methyl-N-(1-methylpropyl)-3-isoquinolinecarboxam ide (PK11195), dipyridamole, or 7-(dimethylcarbamoloxy)-6-phenylpyrrolo-[2,1-d]benzothiazepi ne (DPB), competitive ligands of the mitochondrial benzodiazepine receptor, all diminished protoporphyrin IX accumulation, as did acifluorfen, a mitochondrial protoporphyrinogen oxidase inhibitor. In addition to protoporphyrin IX (Em(max): 630 nm), delta-aminolaevulinic acid-treated cells also generated a fluorophore of Em(max) 580 nm; this compound was identified as Zn-protoporphyrin IX. Mitochondrial benzodiazepine receptor ligands increased the formation of the zinc porphyrin whilst decreasing that of protoporphyrin IX. The involvement of the mitochondrial benzodiazepine receptor in the translocation of porphyrins and the formation of Zn-protoporphyrin IX have wide implications for the use of delta aminolaevulinic acid in photodynamic therapy. PMID- 11020480 TI - Inhibitory effect of various thrombin inhibitors on shear-induced platelet function and dynamic coagulation. AB - We assessed the effects of active site-directed, fibrinogen recognition exosite (FRE)-directed and bifunctional thrombin inhibitors, on shear-induced platelet reactivity (adhesion/aggregation) and dynamic coagulation (coagulation of flowing blood). An in vitro test for shear-induced haemostatic plug formation and dynamic coagulation (haemostatometry) was employed using non-anticoagulated rat blood. The active site-directed inhibitors (argatroban, P891, P899) caused inhibition of platelet reactivity and coagulation at 1-, 100- and 100-microM concentrations, respectively. Bifunctional inhibitors (P553, P1053) exerted inhibitory effects at 0.1 microM. A dimeric bifunctional inhibitor P824 caused significant inhibition at 1 microM. The FRE-directed inhibitor (P960) inhibited shear-induced platelet reactivity at 10 microM but the dynamic coagulation at 1 microM. Combination of active site-directed argatroban and FRE-directed P960 did not show any synergistic effect. The most potent inhibition was observed in monomeric bifunctional inhibitors. The inhibitory effects were compared with the K(i) values against human thrombin and with the IC(50) values against fibrin clot formation. The minimum effective concentrations on shear-induced platelet reactivity and dynamic coagulation were comparable with the IC(50) values, but not with the K(i) values. PMID- 11020481 TI - Neurotoxic/neuroprotective profile of carbamazepine, oxcarbazepine and two new putative antiepileptic drugs, BIA 2-093 and BIA 2-024. AB - We investigated and compared the toxicity profile, as well as possible neuroprotective effects, of some antiepileptic drugs in cultured rat hippocampal neurons. We used two novel carbamazepine derivatives, (S)-(-)-10-acetoxy-10,11 dihydro-5H-dibenz[b, f]azepine-5-carboxamide (BIA 2-093) and 10, 11-dihydro-10 hydroxyimino-5H-dibenz[b,f]azepine-5-carboxamide (BIA 2-024), and compared their effects with the established compounds carbamazepine and oxcarbazepine. The assessment of neuronal injury was made by using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl (MTT) assay, as well as by analysing morphology and nuclear chromatin condensation (propidium iodide staining), after hippocampal neurons were exposed to the drugs for 24 h. The putative antiepileptic drugs, BIA 2-093 or BIA 2-024 (at 300 microM), only slightly decreased MTT reduction, whereas carbamazepine or oxcarbazepine were much more toxic at lower concentrations. Treatment with the antiepileptic drugs caused nuclear chromatin condensation in some neurons, which is characteristic of apoptosis, and increased the activity of caspase-3-like enzymes, mainly in neurons treated with carbamazepine and oxcarbazepine. The toxic effect caused by carbamazepine was not mediated by N methyl-D-aspartate (NMDA) or by alpha-amino-3-hydroxy-5-methyl-isoxazole-4 propionate (AMPA) receptors. Moreover, the antiepileptic drugs failed to protect hippocampal neurons from the toxicity caused by kainate, veratridine, or ischaemia-like conditions. PMID- 11020482 TI - Potentiation of nerve growth factor-action by picrosides I and II, natural iridoids, in PC12D cells. AB - Natural iridoid, picroside I (beta-D-glucopyranoside, 1a,1b,2,5a,6, 6a-hexahydro 6-hydroxy-1a-(hydroxymethyl)oxireno[4,5]cyclopenta[1, 2-c]pyran-2-yl, 6-(3-phenyl 2-propenoate)) or II (beta-D-glucopyranoside, 1a,1b,2,5a,6, 6a-hexahydro-6-[(4 hydroxy-3-methoxybenzoyl)oxy]-1a-(hydroxymethyl )ox ireno[4,5]cyclopenta[1,2 c]pyran-2-yl) alone did not exhibit neuritogenic activity, but caused a concentration-dependent (>0.1 microM) enhancement of nerve growth factor (NGF, 2 ng/ml)-induced neurite outgrowth from PC12D cells. The picroside-induced enhancing action of NGF was abolished by GF109203X (2-[1-(3-dimethylaminopropyl) indol-3-yl]-3-(indol-3-yl)maleimide) (0.1 microM), a protein kinase C inhibitor. Furthermore, PD98059 (2-(2'-amino-3'-methoxyphenyl)-oxanaphthalen-4-one) (20 microM), a potent mitogen-activated protein (MAP) kinase kinase inhibitor, completely blocked the picroside-induced enhancement of neurite outgrowth in the presence of NGF (2 ng/ml), suggesting that picrosides activate the MAP kinase dependent signaling pathway. Interestingly, no increase in the expression of phosphorylated MAP kinase was observed in picroside-treated (60 microM) PC12D cells in the presence of NGF (2 ng/ml). These results suggest that picroside I or II enhances NGF-induced neurite outgrowth from PC12D cells, probably by amplifying a down-stream step of MAP kinase in the NGF receptor-mediated intracellular MAP kinase-dependent signaling pathway. PMID- 11020483 TI - Mitogen-induced up-regulation of non-smooth muscle isoform of alpha-tropomyosin in rat aortic smooth muscle cells. AB - Correlation between the expression of the alpha-tropomyosin isoforms and cell growth was investigated in rat aortic smooth muscle cells. The levels of exon 1a, exons 1a+2a (smooth muscle type) and exons 1a+2b (non-smooth muscle type) were determined by reverse transcription-polymerase chain reaction (RT-PCR). When the cells were cultured, the level of exons 1a+2b transiently increased while reaching a maximum at 3-5 days. When the serum-deprived confluent cells were stimulated with 3-20% serum for 1.5 h, the level of exons 1a+2b increased by about twofold. The 1-(5-isoquinolinesulphonyl)-2-methylpiperazine (H-7) but not 2 [1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)-maleimi de (GF 109203X) inhibited this up-regulation. Phorbol-12, 13-dibutyrate (PDB) mimicked the effect of serum. The DNA synthesis as determined by the incorporation of 5 bromo-2'-deoxy-uridine (BrdU) was not enhanced by the 1.5 h stimulation with serum or phorbol ester. The up-regulation of non-smooth muscle isoform of alpha tropomyosin occurred during G(0)/G(1) transition before entering S phase. Protein phosphorylation is suggested to be involved in the up-regulation. However, the responsible kinase(s) remain to be elucidated. PMID- 11020484 TI - Effect of recombinant human erythropoietin on cerebral ischemia following experimental subarachnoid hemorrhage. AB - Erythropoietin exerts a neuroprotective effect during cerebral ischemia. We investigated the effect of systemic administration of recombinant human erythropoietin in a rabbit model of subarachnoid hemorrhage-induced acute cerebral ischemia. The animals were divided into three groups: group 1, subarachnoid hemorrhage; group 2, subarachnoid hemorrhage plus placebo; group 3, subarachnoid hemorrhage plus recombinant human erythropoietin (each group, n=8). Experimental subarachnoid hemorrhage was produced by injecting autologous blood into the cisterna magna. Treatment with recombinant human erythropoietin and placebo was started 5 min after subarachnoid hemorrhage and was continued every 8 h for 24 h. Before the animals were killed, erythropoietin concentration was measured in the cerebrospinal fluid. The rabbits were killed 24 h after subarachnoid hemorrhage and ischemic brain injury was histologically evaluated. In group 3, the concentration of erythropoietin in the cerebrospinal fluid was significantly increased and a significant reduction in cortical necrotic neuron count was also observed. These findings may encourage the use of erythropoietin in the treatment of cerebral ischemia that often occurs in the early stage of subarachnoid hemorrhage. PMID- 11020485 TI - Pretreatment with morphine potentiates naloxone-conditioned place aversion in mice: effects of NMDA receptor antagonists. AB - Acute pretreatment with opioid receptor agonists potentiates behavioral effects of opioid antagonists. This phenomenon was suggested to serve as an acute model of opioid dependence. Since antagonists acting at N-methyl-D-aspartate (NMDA) receptors were repeatedly shown to attenuate development, maintenance, and expression of opioid dependence, the present study evaluated the effects of competitive NMDA receptor antagonist, D-CPPene (SDZ EAA 494; 3-(2 carboxypiperazin-4-yl)-1-propenyl-1-phosphonic acid), and low-affinity channel blocker, 1-amino-3,5-dimethyl adamantane hydrochloride (memantine), on establishment of naloxone-conditioned place aversion in mice that were pre exposed to morphine. Morphine (20 mg/kg) pretreatment significantly potentiated the ability of naloxone (0.01-0.3 mg/kg) to produce place aversion. The place aversion produced by naloxone (0.1 mg/kg) was attenuated by D-CPPene (1 and 3 mg/kg but not 0.1 or 0.3 mg/kg) when it was administered 3.5 h after morphine (0.5 h prior to conditioning trial with naloxone) but not 0.5 h prior to morphine. Memantine (1-10 mg/kg) had no effect under any treatment condition (0.5 h prior to morphine, simultaneously with morphine, 2 or 3.5 h after morphine). Thus, the ability of NMDA receptor antagonist to affect development and/or expression of morphine dependence may not be a good predictor of their effects on establishment of morphine-potentiated naloxone-conditioned place aversion. PMID- 11020486 TI - Comparison of responses to siguazodan, rolipram, and zaprinast in the feline pulmonary vascular bed. AB - The present study was undertaken to investigate and compare responses to the cyclic nucleotide phosphodiesterase inhibitors siguazodan (type III, guanosine 3',5'-cyclic monophosphate (cGMP)-inhibited adenosine 3',5'-cyclic monophosphate (cAMP)), rolipram (type IV, cAMP-specific), and zaprinast (type V, cGMP-specific) in the feline pulmonary vascular bed. When tone in the pulmonary vascular bed was raised to a high steady level with a constant infusion of the thromboxane mimic U46619 (9,11-dideoxy-11, alpha9alpha-epoxymethano prostaglandin F(2alpha)), intralobar injections of the three phosphodiesterase inhibitors caused dose related decreases in lobar arterial pressure. In terms of relative vasodilator activity, rolipram was more potent at higher doses than siguazodan, which was more potent than zaprinast. The duration of the pulmonary vasodilator response to zaprinast was shorter than for siguazodan or rolipram. Furthermore, siguazodan and rolipram, but not zaprinast, decreased systemic arterial pressure when injected into the perfused lobar artery in the range of doses studied. The present data demonstrate that the three phosphodiesterase inhibitors have potent, long-lasting vasodilator activity in the pulmonary vascular bed of the cat. These data suggest that there is rapid turnover of cAMP and cGMP in the pulmonary circulation and indicate that phosphodiesterase enzyme types III, IV, and V may play an important role in the regulation of vasomotor tone in the feline lung. PMID- 11020487 TI - Role of phospholipase A(2) and myoendothelial gap junctions in melittin-induced arterial relaxation. AB - We have used preconstricted rings of rabbit superior mesenteric artery to investigate the contribution of phospholipase A(2) and gap junctional communication to endothelium-derived hyperpolarizing factor (EDHF)-type relaxations evoked by melittin, a polypeptide toxin known to mobilize arachidonic acid from the cell membrane. Arachidonyl trifluoromethyl ketone (30 microM), an inhibitor of the Ca(2+)-dependent phospholipase A(2), and Gap 27 (300 microM), a connexin-mimetic peptide which attenuates intercellular communication via gap junctions, both abolished the endothelium-dependent component of EDHF-type responses evoked by melittin in the presence of the NO synthase inhibitor N(G) nitro-L-arginine methyl ester (L-NAME, 300 microM) and the cyclooxygenase inhibitor indomethacin (10 microM). By contrast, the sulfhydryl agent thimerosal (300 nM), which amplifies EDHF activity, potentiated nitric oxide (NO)/prostanoid independent relaxations induced by melittin. Neither arachidonyl trifluoromethyl ketone nor thimerosal modulated relaxations evoked by the peptide toxin in the absence of L-NAME and indomethacin. We conclude that melittin evokes EDHF-type relaxations through activation of the endothelial Ca(2+)-dependent phospholipase A(2) followed by the transmission of a chemical and/or electrical signal via myoendothelial gap junctions. This mechanism of vasorelaxation may be negatively regulated by NO. PMID- 11020488 TI - Differences in the nitric oxide/soluble guanylyl cyclase signalling pathway in the myocardium of neonatal and adult rats. AB - The effects of a nitric oxide-donor, S-nitroso-N-acetylpenicillamine, and a direct activator of soluble guanylyl cyclase, 3-(5'-hydroxymethyl-2'-furyl)-1 benzyl indazole (YC-1), on force of contraction (F(c)) and L-type Ca(2+) currents (I(Ca(L))) were investigated in myocardial preparations from neonatal and adult rats. Since hearts from adult and neonatal animals contained 160 and 47 mg/100 g wet weight myoglobin, respectively, its possible interaction with both drugs was also investigated. Both S-nitroso-N-acetylpenicillamine (100 microM) and YC-1 (30 microM) were ineffective in myocardial preparations from adult rats but reduced the magnitude of I(Ca(L)) and F(c) in preparations from neonatal rats. The latter effects were antagonised by 1H-[1,2, 4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 50 microM) and attenuated by myoglobin (30-300 microM), which also attenuated the effects of both drugs on pre-contracted aortic rings. The differential effects of S-nitroso-N-acetylpenicillamine and YC-1 in the myocardium from adult and neonatal rats may result from developmental changes in the content of myoglobin and/or in the NO/soluble guanylyl cyclase signal pathway. PMID- 11020489 TI - Cardiac and vascular effects of nitric oxide synthase inhibition in lipopolysaccharide-treated rats. AB - In the present study, intraperitoneal injection of lipopolysaccharide (10 mg/kg) to anaesthetized rats produced a gradual fall in mean arterial pressure in 6 h. Aortic rings from lipopolysaccharide-treated rats showed a significant reduction in the contractile response to vasoconstrictors. Pretreatment with N(G)-nitro-L arginine methyl ester (L-NAME) or aminoguanidine, two nitric oxide synthase (NOS) inhibitors, abolished this vascular hyporeactivity. In ventricular myocytes isolated from lipopolysaccharide-treated rats, both electrically induced Ca(2+) transients and the intracellular Ca(2+) response to beta-adrenergic stimulation were significantly depressed when compared with those recorded from myocytes from sham control rats. L-NAME and aminoguanidine alone had no effects on electrically stimulated Ca(2+) transients in ventricular myocytes either from control or lipopolysaccharide-treated rats. However, these two NOS inhibitors augmented the intracellular Ca(2+) response to beta-adrenergic stimulation in myocytes from lipopolysaccharide-treated rats, but not in control myocytes. In addition, 1H [1,2,4]oxadiazolo[4, 3-a]quinoxalin-1-one (ODQ), an inhibitor of nitric oxide (NO)-sensitive guanylyl cyclase, also reversed the intracellular Ca(2+) hyporesponsiveness to beta-adrenergic stimulation in myocytes from lipopolysaccharide-treated rats. In cardiac myocytes from lipopolysaccharide-rats pretreated with aminoguanidine, the intracellular Ca(2+) hyporesponsiveness to beta-adrenergic stimulation was abolished. However, there still existed a depressed Ca(2+) response to electrical field stimulation. These data indicate that NO following lipopolysaccharide stimulation contributes to vascular hyporeactivity and the depressed intracellular Ca(2+) response to beta-adrenergic stimulation in lipopolysaccharide-treated rats, but is not responsible for the reduced Ca(2+) response to electrical stimulation in our experimental conditions. PMID- 11020490 TI - Human urotensin II mediates vasoconstriction via an increase in inositol phosphates. AB - The cyclic peptide urotensin II has recently been cloned from human and reported to potently constrict primate blood vessels. To elucidate the cellular signalling mechanisms of this peptide, we investigated a possible relationship of vasomotor effects of human urotensin II and phosphoinositide turnover in isolated rabbit thoracic aorta. Human urotensin II produced a slowly developing increase in isometric contractile force (pEC(50)=9.0) that was endothelium-independent. The contractile effect of urotensin II was significantly inhibited by the phospholipase C inhibitor, 2-nitro-4-carboxyphenyl-N,N,-diphenylcarbamate (NCDC), but not by the cyclooxygenase inhibitor, indomethacin. In slices of rabbit thoracic aorta, human urotensin II increased phosphoinositide hydrolysis, and this effect was also inhibited by NCDC. The potency of urotensin II (pEC(50)=8.6) was similar to that found in the contractile studies. Thus, vasoconstrictor effects of human urotensin II appear to be mediated by a phospholipase C dependent increase in inositol phosphates, suggesting that the peptide acts via a G(q) protein-coupled receptor. PMID- 11020491 TI - Evaluation of Y-27632, a rho-kinase inhibitor, as a bronchodilator in guinea pigs. AB - To evaluate (+)-(R)-trans-4-(l-Aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride, monohydrate (Y-27632), a selective Rho-kinase inhibitor, as a novel bronchodilator in vivo and in vitro, we investigated the effect of Y-27632 on the acetylcholine- or ovalbumin-induced increase in lung resistance (R(L)) in non-sensitized or passively sensitized guinea pigs, and the relaxant effects of salbutamol, Y-27632 and theophylline on acetylcholine- or ovalbumin-induced contraction of isolated trachea. Y-27632 inhalation (1 mM, 2 min) inhibited acetylcholine- or ovalbumin-induced increase in R(L) without changes in mean blood pressure, and the effect persisted for at least 3 h. Salbutamol, Y-27632 and theophylline each completely reversed the acetylcholine- or ovalbumin-induced contraction of isolated trachea with rank order of potency, salbutamol>Y 27632>theophylline. The relaxant effect of Y-27632 was not affected by propranolol. We conclude that, although Y-27632 is not as potent as a beta adrenoceptor agonist, Y-27632 may become an alternative inhaled bronchodilator, because Y-27632 is more potent than theophylline, and the relaxant effect is independent of beta-adrenoceptors. PMID- 11020492 TI - S-15176 reduces the hepatic injury in rats subjected to experimental ischemia and reperfusion. AB - The protective effect of N-[(3, 5-di-tertiobutyl-4-hydroxy-1-thiophenyl)]-3 propyl-N'-(2,3, 4-trimethoxybenzyl)piperazine (S-15176) on liver injury induced by warm ischemia-reperfusion was investigated using a rat model. Animals were subjected to 2 h of ischemia followed by different reperfusion times. Hepatocyte integrity was assessed by measuring plasma alanine and aspartate aminotransferase activities, and by determining reduced and oxidized glutathione in plasma and bile. Hepatocyte function was quantitated by determining bile flow and liver ATP content. Ischemia-reperfusion resulted in severe hepatic injury involving a huge increase in alanine and aspartate aminotransferase activities, a drop in ATP content, and a decrease in bile flow. Plasma and bile reduced (GSH) and oxidized (GSSG) glutathione concentrations were inversely related: plasma levels increased when biliary levels decreased. This was associated with a decrease in animal survival (-34%). S-15176 pretreatment (1.25, 2.5, 5 or 10 mg kg(-1) day(-1)) improved the survival rate and limited tissue damages in a dose-dependent manner. The pretreatment also reduced the aminotransferase leakage from hepatocytes and the increase in plasma glutathione levels. In addition, normalization of the plasma GSSG/GSH ratio, a good index of an oxidative stress, was observed in groups treated with the higher dosage, suggesting that the antioxidant properties demonstrated for the compound in vitro (IC(50)=0.3 microM towards lipid peroxidation) could play a role in its protective effect. S-15176 pretreatment also protected the organ from the drop in ATP levels. At the higher dose, ATP content was maintained at a level almost 86% of the sham-operated group after 60 min of reperfusion. This was associated with a restoration of the biliary flow. These data suggest that S-15176 may be a useful drug in liver surgery to prevent ischemia-reperfusion injury. PMID- 11020493 TI - Effect of neocuproine, a selective Cu(I) chelator, on nitrergic relaxations in the mouse corpus cavernosum. AB - The effects of neocuproine and bathocuproine, Cu(I) and Cu(II) chelators, respectively, were studied on relaxations in response to electrical field stimulation, acetylcholine, S-nitrosoglutathione, acidified sodium nitrite and sodium nitroprusside in the mouse corpus cavernosum precontracted with phenylephrine. Neocuproine significantly inhibited relaxations induced by electrical field stimulation, acetylcholine and S-nitrosoglutathione, but not by acidified sodium nitrite and sodium nitroprusside. The pre-prepared neocuproine Cu(I) complex was ineffective on the responses. The discrepancy between the shape of relaxations in response to electrical field stimulation or to acetylcholine and S-nitrosoglutathione was abolished by adding CuCl(2) into the bathing medium. The copper action was blocked by neocuproine but not by bathocuproine. However, the pre-prepared bathocuproine-Cu(II) complex did not accelerate the relaxations affected by CuCl(2). These findings suggest that a Cu(I)-dependent mechanism may play a role in the relaxation induced by the endogenous relaxant factor as well as by S-nitrosoglutathione in mouse cavernosal tissue. PMID- 11020494 TI - Less is more: the risks of multiple births. The Institute for Science, Law, and Technology Working Group on Reproductive Technology. AB - OBJECTIVE: To review the medical, social, and financial risks caused by the birth of multiples that need to be addressed in policy and practice. RESULT(S): Many risks of multiple births are described in the literature. The medical risks to the offspring include death, low birth weight, deformational plagiocephaly, and other physical and mental disabilities. Risks to the women include premature labor, premature delivery, pregnancy-induced hypertension, toxemia, gestational diabetes, and vaginal-uterine hemorrhage. Children born in multiples face difficulty socializing, developmental delays, and behavioral problems, whereas their parents risk exhaustion, depression, and anxiety. In addition to personal costs faced by families, society often bears the financial costs of overburdened hospitals, caps on insurance and/or inability of parents to cover expenses. CONCLUSION(S): Multiple births present potential acute and long-term medical risks to the pregnant woman and her children. However, more long-term follow-up research and more research on outcomes with higher-order multiples are needed. In designing practices and policies to improve the success of IVF while reducing the risk of multiples, it is important to balance the many interests involved. At a minimum, providers and patients need to be educated about the risks of multiple gestation so that steps can be taken to prevent adverse outcomes. PMID- 11020496 TI - Follicle pool depletion: factors involved and implications. PMID- 11020495 TI - Predictors of ovarian follicle number. AB - OBJECTIVE: To identify factors associated with ovarian follicle density in women of reproductive age. DESIGN: Women undergoing incidental oophorectomy responded to structured interviews. Grossly normal ovaries were sectioned and assessed for follicle density. SETTING: University hospital. PATIENT(S): 102 women undergoing hysterectomy with incidental oophorectomy who had morphologically normal ovaries. MAIN OUTCOME MEASURE(S): Follicle density, age, parity, oral contraceptive use, body size, smoking, and alcohol consumption. RESULT(S): Follicle density decreased with age and with current or past cigarette smoking. Other factors were not associated with follicle density, although the study had adequate power to detect only strong associations. CONCLUSION(S): Some of the adverse effects of smoking on reproductive health may be due to irreversible follicular depletion. PMID- 11020497 TI - Antiphospholipid antibodies in patients undergoing IVF: the data do not support testing. PMID- 11020498 TI - "It's not what you don't know that makes you look like a fool. it's what you do know that ain't so."--Appalachian Mountain proverb. PMID- 11020499 TI - Deja vu all over again. PMID- 11020500 TI - Assisted reproductive technology in the United States: 1997 results generated from the American Society for Reproductive Medicine/Society for Assisted Reproductive Technology Registry. AB - OBJECTIVE: To summarize the procedures and outcomes of assisted reproductive technology (ART) initiated in the United States in 1997. DESIGN: Data were collected electronically by using Society for Assisted Reproductive Technology Clinical Outcome Reporting System software and were submitted to the American Society for Reproductive Medicine/Society for Assisted Reproductive Technology Registry. PARTICIPANT(S): 335 programs submitted data on procedures performed in 1997. Data were collated after November 1998 so that the outcome of all pregnancies established would be known. MAIN OUTCOME MEASURE(S): Incidence of clinical pregnancy, ectopic pregnancy, abortion, stillbirth, delivery, and structural and functional abnormalities. RESULT(S): Programs reported initiating 73,069 cycles of ART treatment. Of these, 51,344 cycles involved IVF (with and without micromanipulation), with a delivery rate per retrieval of 27.9%; 1,943 were cycles of GIFT, with a delivery rate per retrieval of 30.0%; and 1,104 were cycles of zygote intrafallopian transfer, with a delivery rate per retrieval of 28.0%. The following additional ART procedures were also initiated: 4,616 donor oocyte cycles, with a delivery rate per transfer of 40.0%; 10,181 frozen embryo transfer procedures, with a delivery rate per transfer of 18.8%; 1,584 frozen embryo transfers using donated oocytes, with a delivery rate per transfer of 22.2%; and 600 cycles using a host uterus, with a delivery rate per transfer of 34.6%. Furthermore, 1,173 cycles were reported as combinations or more than one treatment type, 40 cycles as research, 258 as embryo banking, and 226 as other (unclassified) cycle types. As a result of all procedures, 17,311 deliveries resulting in 25,059 babies were reported. CONCLUSION(S): In 1997, more programs reported ART treatment and the number of reported cycles increased significantly (10.9%) compared with 1996. In comparable cycle types, the overall success rate (deliveries per retrieval) increased by 1.8%, which represents an increase of 6.9% compared with the success rate for 1996. PMID- 11020501 TI - Commentary PMID- 11020502 TI - Should we still advise infertile couples to use (barrier) contraception before IVF down-regulation? AB - OBJECTIVE: To determine the outcome of spontaneous conceptions in women who received GnRH agonists during mid-luteal phase down-regulation before IVF treatment. DESIGN: Retrospective analysis of case records and study of the literature. SETTING: Two university-affiliated reproductive medicine units. PATIENT(S): Seventy-three women who conceived spontaneously after starting down regulation with a GnRH agonist before controlled ovarian hyperstimulation. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Course and clinical outcome of pregnancies. RESULT(S): Seventy-four pregnancies occurred in 73 women who received a GnRH agonist. Of these patients, 6 (8%) had a biochemical pregnancy, 6 (8%) had an ectopic pregnancy, 21 (28%) miscarried, and 41 pregnancies resulted in successfully delivered babies; there were 2 cases of congenital abnormalities. CONCLUSION(S): These cases, together with other published data, suggest that pregnancy outcome is not adversely affected by exposure to GnRH agonist during luteal-phase down-regulation. A central register of pregnant women who received a GnRH agonist is needed. PMID- 11020503 TI - A follow-up study with oocyte donors exploring their experiences, knowledge, and attitudes about the use of their oocytes and the outcome of the donation. AB - OBJECTIVE To learn what information oocyte donors were given and wanted to have about the use of their oocytes and the outcome of the donation. DESIGN: In-depth interviews. SETTING: Participants recruited through IVF clinics, matching agency, the Internet, word of mouth, and newspaper ads. PARTICIPANT(S): Thirty-three former oocyte donors and six women preparing to donate. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): None. RESULT(S): Thirty-three former donors completed 66 donation cycles; 48 donation cycles were anonymous. Only 41% (16 of 39) of all participants were comfortable giving the recipient couple complete dispositional authority over the resulting embryos; the remainder wanted some control. One quarter did not want embryos used for research. Fifty-four percent (21 of 39) thought donation of excess embryos to another couple was acceptable, but one third wanted to be informed. Of the 25 anonymous donors, 6 learned the outcome of the donation; 14 others wanted to know. All hoped the donation was successful. CONCLUSION(S): Because oocyte donors' need for information varies, clinics should consider being more flexible in their disclosure policies. Disclosure about the possible uses of donor oocytes or embryos should be mandatory. Findings have implications for the informed consent and counseling processes. PMID- 11020504 TI - Initial IVF-ET experience with assisted hatching performed 3 days after retrieval followed by day 5 embryo transfer. AB - OBJECTIVE: To report our initial IVF-ET experience combining assisted hatching performed 3 days after oocyte retrieval with day 5 embryo transfer (ET). DESIGN: Retrospective review of 110 consecutive IVF cycles not involving donor oocytes, including 16 cycles that involved assisted hatching performed 3 days after oocyte retrieval in combination with day 5 ET. SETTING: Academic teaching hospital IVF center. PATIENT(S): Eighty-six consecutive IVF patients undergoing ET. INTERVENTION(S): Assisted hatching using acid Tyrode's solution performed 3 days after oocyte retrieval in selected cases in combination with day 3 or 5 ETs. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate per ET. RESULT(S): Of the 16 women undergoing day 5 ET following day 3 assisted hatching, 14 had a clinical pregnancy. These included 11 ongoing/delivered singletons and 2 ongoing/delivered twin pregnancies, neither of which was monochorionic. These clinical and ongoing/delivered pregnancy rates compared very favorably with those of 54% and 46%, respectively, for the 35 patients undergoing day 5 ETs without assisted hatching, even though the latter group appeared to be better IVF candidates based on the prognostic factors commonly used to predict success. CONCLUSION(S): Our experience suggests that day 3 assisted hatching followed by day 5 ET may be a useful combination in selected patients. Although not seen in our small series, an increased risk of monochorionic pregnancies remains a theoretical concern when such a combination is used, since both assisted hatching and blastocyst transfers have been independently linked to an increased risk in some reports. PMID- 11020505 TI - Effect of chromosomal translocations on the development of preimplantation human embryos in vitro. AB - OBJECTIVE: To determine the reliability of a new technique for single human blastomere karyotyping during clinical cases for preimplantation genetic diagnosis of translocations. DESIGN: Controlled clinical study. SETTING: Preimplantation genetic diagnosis and IVF program. PATIENT(S): Nineteen preimplantation genetic diagnosis cases with 11 types of translocations (10 reciprocal and one Robertsonian) involving chromosomes 1, 5, 7, 8, 9, 11, 12, 13, 14, 15, 16, 18, 20, 21, and 22. INTERVENTION(S): Blastomere biopsy followed by blastomere nucleus conversion into metaphase chromosomes. Fluorescent in situ hybridization (whole chromosome painting) was used for the detection of chromosomally unbalanced preimplantation human embryos. MAIN OUTCOME MEASURE(S): Percentage of informative metaphase plates and effect of unbalanced translocations on preimplantation embryo development. RESULT(S): Informative metaphases were obtained for 84% of the blastomeres. Analysis of preimplantation development of the resulting embryos showed that an unbalanced chromosomal complement does not affect embryo ability to reach the blastocyst stage in vitro. CONCLUSION(S): For the translocations tested, there is no evident selection against chromosomally unbalanced embryos at the preimplantation stage of embryo development. PMID- 11020506 TI - The choice of embryo transfer catheter affects embryo implantation after IVF. AB - OBJECTIVE: Comparison of two transfer catheters in an IVF program. DESIGN: Prospective, randomized clinical study. SETTING: A private tertiary care center for ART. PATIENT(S): 66 patients < 38 years of age undergoing IVF and/or ICSI. INTERVENTION(S): Patients were randomly assigned to undergo ET using the Tomcat catheter (n = 32) or the TDT catheter (n = 34). MAIN OUTCOME MEASURE(S): Primary outcome measures were implantation and pregnancy rates. Secondary outcome measures were contamination with blood and/or mucus on the tip of the catheter, cramping or patient discomfort, and time required to complete ET. RESULT(S): Use of the Tomcat catheter resulted in significantly higher implantation (25.2% vs. 8.4%) and clinical pregnancy rates (47% vs. 14.7%) compared with the TDT catheter. All secondary outcome measures were similar for both catheters. CONCLUSION(S): The choice of ET catheter may affect the success of IVF-ET cycles. Use of the Tomcat catheter compared with the TDT catheter seems to result in significantly better efficiency of the ET procedure and is more cost effective. PMID- 11020507 TI - Analysis of 104 twin pregnancies conceived with assisted reproductive technologies and 193 spontaneously conceived twin pregnancies. AB - OBJECTIVE: To evaluate pregnancy outcome of assisted reproductive technology (ART)-conceived twin pregnancies. DESIGN: Retrospective study. SETTING: A tertiary obstetric care center. PATIENT(S): All twin pregnancies delivered > or = 24 weeks of gestation from January 1, 1996, to December 31, 1997. INTERVENTION(S): Maternal and neonatal record review. MAIN OUTCOME MEASURE(S): Pregnancy and perinatal outcome. RESULT(S): The study group comprised 104 ART conceived twin pregnancies, and 193 non-ART-conceived pregnancies served as controls. Mean maternal age, the proportion of nulliparae, and the percentage of women who delivered before 34 weeks' gestation was higher among the study women, whereas mean gestational age was younger. The incidences of pregnancy-induced hypertension, uterine bleeding, premature contractions, intrauterine growth retardation, fetal death, discordance, and cesarean section were significantly higher in the study group. Correspondingly, in the study group, the mean birth weight of both twins was lower; more neonates weighed < 1, 500 g, more had Apgar scores of < 7 at 5 minutes, more were admitted to the intensive care unit, and more second twin neonates died. The outcome of twin pregnancies conceived spontaneously was comparable with those conceived by ovulation induction. CONCLUSION(S): Assisted reproductive technology-conceived twin pregnancies are at greater risk than non-ART-conceived ones for pregnancy complications and adverse perinatal outcome. PMID- 11020508 TI - Paternal occupational exposures and embryo implantation rates after IVF. AB - OBJECTIVE: To investigate any influence of paternal occupational exposures on implantation rates after IVF. DESIGN: Cohort study of couples who sought IVF treatment. SETTING: University Hospital Utrecht, The Netherlands. PATIENT(S): The study population was composed of 726 couples pursuing IVF treatment. INTERVENTION(S): Only the earliest IVF treatment cycle with ET was selected for the analysis. All couples filled in a generic questionnaire on lifestyle factors and details about their occupation. In addition, more detailed exposure information was obtained for pesticides with use of job-specific questionnaires and a subsequent telephone interview. MAIN OUTCOME MEASURE(S): The implantation rate was defined as the number of gestational sacs seen with ultrasound at 6-7 weeks of pregnancy, divided by the number of embryos replaced. RESULT(S): A significantly reduced implantation rate was seen among couples with male partners working in occupations with presumably high levels of organic solvent exposure. Conversely, paternal pesticide exposure was significantly associated with an increased implantation rate. Paternal exposures to metal dust or fumes and welding fumes were not related to the probability of implantation. CONCLUSION(S): The findings suggested that paternal organic solvent exposure decreased the implantation rate among couples undergoing IVF-ET treatment. PMID- 11020509 TI - Use of electively cryopreserved microsurgically aspirated epididymal sperm with IVF and intracytoplasmic sperm injection for obstructive azoospermia. AB - OBJECTIVE: To investigate the efficacy of using intentionally cryopreserved epididymal sperm in selected cases of obstructive azoospermia. DESIGN: A retrospective, nonrandomized study. SETTING: Academic research environment. PATIENTS: One hundred forty-one couples undergoing first-time IVF/ICSI using either fresh or cryopreserved epididymal sperm. INTERVENTIONS: The epididymides were microsurgically aspirated. MAIN OUTCOME MEASURES: Clinical pregnancy rates. RESULTS: Motile sperm were obtained from all men. For the fresh group, the mean total sperm aspirated was 99 x 10(6) with 5.5 vials frozen per patient after ICSI and 82 x 10(6) with 4.7 vials frozen per patient in the cryopreserved group. No statistically significant difference in oocyte fertilization rate or number of embryos transferred was noted between groups. Of 108 patients using freshly aspirated sperm, 72 (66.7%) achieved clinical pregnancy. Of 33 patients in the group using cryopreserved sperm, 20 (60.6%) achieved clinical pregnancy (P=0.47). CONCLUSIONS: In selected ideal cases of unreconstructable azoospermia, elective open microsurgical epididymal sperm aspiration with cryopreservation yields pregnancy rates similar to that employing fresh sperm. The advantages of this method are: (1) Use of cryopreserved sperm obviates the logistics problems associated with the use of fresh sperm. (2) Abundant high-quality sperm can be cryopreserved in a single procedure for all future attempts at IVF/ICSI. Rarely, viable sperm will not be present after thawing, and fresh retrieval will be necessary. PMID- 11020510 TI - Psychological well-being and coping patterns in infertile men. AB - OBJECTIVE: To determine whether differences existed in mood and coping styles among fertile men, oligoasthenospermic men, or euspermic men whose wives were undergoing ovulation stimulation with clomiphene and IUI. DESIGN: A cross sectional research design. SETTING: Hospital-based academic fertility clinic. PATIENT(S): 30 fertile men with currently pregnant wives, 30 euspermic and 30 oligoasthenospermic men in couples undergoing ovulation stimulation with clomiphene and IUI. INTERVENTION(S): Measures of psychological well-being and coping were administered. MAIN OUTCOME MEASURE(S): Biodemographic information, and psychometric measures of mood and coping. RESULT(S): There were no significant differences among the groups on any of the measures except the Family Inventory of Life Events (FILE), in which fertile men reported higher stress levels. FILE scores in all groups were moderate, indicating typical levels of family stress. CONCLUSION(S): Mood and coping in the three groups were similar. This study suggests that men's psychological adjustment to their own infertility and to unexplained infertility is generally healthy. PMID- 11020511 TI - Desogestrel plus testosterone effectively suppresses spermatogenesis but also causes modest weight gain and high-density lipoprotein suppression. AB - OBJECTIVE: To test the hypothesis that oral desogestrel (DSG) plus testosterone would uniformly and rapidly suppress sperm concentrations in young men as effectively as levonorgestrel (LNG) plus testosterone and cause less high-density lipoprotein (HDL) suppression and weight gain. DESIGN: Single-blind, randomized trial. SETTING: VA Puget Sound and University of Washington, Seattle, Washington. PATIENT(S): Twenty-four healthy young men, aged 20-49. INTERVENTION(S): Subjects were randomized to three groups of men who were administered 6 months of therapy with oral DSG plus im testosterone enanthate: 150 microg of DSG plus 50 mg of testosterone (DSG 150-T 50), 150 microg of DSG plus 100 mg of testosterone (DSG 150-T 100) or 300 microg of DSG plus 100 mg of testosterone (DSG 300-T 100). We compared these three groups to two groups of historical controls of 100 mg of im testosterone alone or 150 microg of oral LNG plus 100 mg of im testosterone (LNG 125-T 100 group) enrolled in similar studies. MAIN OUTCOME MEASURE(S): Suppression of sperm counts to severe oligoazoospermia (sperm counts <1 x 10(6)/mL) and azoospermia, weight gain, and serum high-density cholesterol (HDL) suppression. RESULT(S): Azoospermia was achieved in all eight men receiving DSG 150-T 100 and seven of the eight men in the DSG 300-T 100 group. DSG 150 or 300 plus T 100 suppressed spermatogenesis as effectively as LNG 125-T 100 and more effectively than DSG 50-T 100 or testosterone alone. All groups tended to gain weight compared with their baseline, but the weight gain was greatest (and statistically significant) in the DSG 150-T 100, DSG 300-T 100, and LNG 125-T 100 groups. Serum HDL levels were modestly suppressed in all groups, and this effect was greatest in the DSG 300-T 100 and LNG 125-T 100 groups. CONCLUSION(S): The combination of DSG plus testosterone is a very effective regimen for suppression of spermatogenesis and has acceptably low side effects. PMID- 11020512 TI - Semen treatment with progesterone and/or acetyl-L-carnitine does not improve sperm motility or membrane damage after cryopreservation-thawing. AB - OBJECTIVE: To assess the effects of progesterone and acetyl-L-carnitine used before semen cryopreservation-thawing on sperm motility parameters and plasma membrane integrity. DESIGN: Prospective cohort study. SETTING: Academic tertiary center. PATIENT(S): Subfertile men undergoing semen evaluation. INTERVENTION(S): Before cryopreservation, spermatozoa were incubated with water-soluble progesterone (1 and 10 microM), acetyl-L-carnitine (2.5, 5, 10, and 20 mM), or both (progesterone, 1 microM; and acetyl-L-carnitine, 5 mM). MAIN OUTCOME MEASURE(S): Postthaw change of motility parameters (computer-assisted measurements) and vitality-membrane integrity (examined with eosin-Y staining and annexin V-Cy3 binding assay). RESULT(S): There were no statistically significant differences between control samples and samples treated with progesterone and/or acetyl-L-carnitine for cryosurvival rate, motility parameters, or membrane integrity. The percentages of postthaw cells identified as live showed significantly different results with use of the eosin-Y staining and annexin V binding assay. CONCLUSION(S): Neither progesterone nor acetyl-L-carnitine seemed to prevent cryodamage assessed by motility changes or membrane integrity in human spermatozoa of subfertile men. Annexin V binding, a reflection of membrane translocation of phosphatidylserine, provided more distinct information about postfreezing membrane integrity changes than eosin-Y staining. PMID- 11020513 TI - A continuous quality control program for strict sperm morphology. AB - OBJECTIVE: To develop a training program with intervals of continuous quality control assessments for the evaluation of strict sperm morphology. DESIGN: Prospective analytical study. SETTING: Academic hospital and academic institution setting. PATIENT(S): Healthy sperm donors. INTERVENTION(S): Nine individual andrology laboratories in Switzerland were invited to participate in a training course for strict sperm morphology, which was followed up every 3 months by a continuous quality control program. Each laboratory received six slides over a period of 9 months, during which time the results were forwarded to the reference laboratory. Papanicolaou stain sperm slides were prepared and shipped to participating laboratories every 3 months. MAIN OUTCOME MEASURE(S): Percentage of normal spermatozoa. RESULT(S): The mean (+/-SE) percentage of normal sperm reported by the reference laboratory compared with the participating laboratories for slides 1-6 were 11. 4 +/- 1.6 vs. 17.3 +/- 6 (P>.2), 6.0 +/- 1.3 vs. 8.6 +/- 2.5 (P>.2), 9.0 +/- 0 vs. 9.6 +/- 3 (P>.2), 1 +/- 0 vs. 1.2 +/- 0.2 (P>.2), 23.3 +/- 0.3 vs. 28.0 +/- 1.3 (P>.2), and 2.0 +/- 0 vs. 6.1 +/- 1.2 (P>.2), respectively. Technician proficiency was reported to differ by <10% from the reference laboratory in 94% of cases. CONCLUSION(S): The results illustrate that training and proficiency testing can be conducted on a national and international level with the support of a reference laboratory. Global quality control measurements in andrology laboratories should become mandatory, since these results indicate that continuous quality control for laboratory technicians can be successful. PMID- 11020514 TI - Delayed conception and active and passive smoking. The Avon Longitudinal Study of Pregnancy and Childhood Study Team. AB - OBJECTIVE: To determine whether passive as well as active smoking by women or smoking by men is associated with delayed conception, after adjustment for confounding factors. DESIGN: Population study of couples expecting a baby. Logistic regression was performed to identify factors associated with delayed conception. SETTING: The Avon Health Authority area, United Kingdom. PATIENT(S): All couples expected to deliver between April 1991 and December 1992. INTERVENTION(S): Questionnaires administered early in pregnancy. MAIN OUTCOME MEASURE(S): Time taken to conceive, categorized as <6 months, 6-11 months, 1-3 years, and >3 years. RESULT(S): After correction for confounding factors, delayed conception was statistically significantly associated with both active smoking by the woman (odds ratio [OR] 1.23 [95% CI 0.98-1.49] for > 6 months and 1.54 [95% CI 1.19-2.01] for >12 months) and her exposure to passive smoking (OR 1.17 [95% CI 1.02-1.37] and 1.14 [95% CI 0.92-1.42]) compared with women with no exposure to tobacco smoke (referent). Heavy smoking by men was independently associated with delayed conception. In active smokers, the effect increased with the number of cigarettes. CONCLUSION(S): Smoking by men and passive and active smoking by women are associated with delayed conception. PMID- 11020515 TI - Oral contraceptive use by teenage women does not affect peak bone mass: a longitudinal study. AB - OBJECTIVE: To determine the effect of oral contraceptive pill (OCP) use during adolescence on peak bone mass. DESIGN: Longitudinal observational study. SETTING: Academic clinical research center. PATIENT(S): Sixty-two non-Hispanic, white females in The Penn State Young Women's Health Study, who were studied for 8 years during ages 12-20. INTERVENTION(S): There were 28 OCP users, who used OCPs for a minimum of 6 months and were still using at age 20, and 34 nonusers who had never used OCPs. MAIN OUTCOME MEASURE(S): Total body bone, dedicated hip bone, and body composition measurements were made by dual-energy roentgenogram absorptiometry. RESULT(S): The OCP users and nonusers did not differ at entry in anthropometric, body composition, or total body bone measurements. By age 20, the average duration of OCP use by the user group was 22 months. At age 20, the groups remained indistinguishable in anthropometric, body composition, total body, and hip bone measures, and in age of menarche and sports exercise scores. CONCLUSION(S): Oral contraceptive pill use by healthy, white, teenage females does not affect acquisition of peak bone mass. PMID- 11020516 TI - Atypical endometriosis revisited: clinical and biochemical evaluation of the different forms of superficial implants. AB - OBJECTIVE: To investigate the activity of different forms of endometriosis implants by clinical and biochemical evaluation. DESIGN: Prospective, blinded study. SETTING: Tertiary-care university hospital. PATIENT(S): Forty-seven infertile patients with minimal or mild endometriosis diagnosed at laparoscopy were included in the clinical investigation. INTERVENTION(S): Patients were submitted to a preoperative evaluation of endometriosis-associated dysmenorrhea by means of a 10-point linear analog scale and to laparoscopic staging of endometriosis following a current classification system. In the biochemical investigation, tissue samples from different endometriosis lesions and control tissues were assessed for prostaglandin (PG) F(2 alpha) production. MAIN OUTCOME MEASURE(S): Evaluation of the correlation between endometriosis-associated dysmenorrhea and the extent of the disease. Evaluation of the production of PGF(2 alpha) by the different tissues sampled. RESULT(S): No positive correlation was present between any of the implant subgroups and the associated dysmenorrhea. White peritoneal implants were associated with milder pain symptoms than black or red lesions. The biologic activity of red and black superficial implants, expressed as the production of PGF(2 alpha), was similar. CONCLUSION(S): No positive correlation was demonstrated between endometriosis-associated dysmenorrhea and the current classification of endometriosis, which includes characterization of the different morphologic aspects of superficial endometriosis implants. PMID- 11020517 TI - Ovarian transposition for patients with cervical carcinoma treated by radiosurgical combination. AB - OBJECTIVE: To assess the indications, effectiveness, and complications of ovarian transposition before pelvic irradiation for cervical cancer. DESIGN: Prospective study. SETTING: Gynecologic oncology department at a French cancer center. PATIENT(S): One hundred seven patients treated for cervical cancer. INTERVENTION(S): Ovarian transposition to the paracolic gutters with radical hysterectomy and lymphadenectomy. MAIN OUTCOME MEASURE(S): Clinical and laboratory follow-up tests for ovarian function. RESULT(S): Bilateral ovarian transposition was achieved in 104 patients (98%). Twelve patients were lost to follow-up or excluded because of evolution of the disease. Preservation of ovarian function was achieved in 83% of the patients having follow-up. The rates of ovarian preservation were 100% for patients treated exclusively by surgery, 90% for patients treated by postoperative vaginal brachytherapy, and 60% for patients treated by postoperative external radiation therapy and vaginal brachytherapy. The main risk for ovarian failure was found in patients treated by external radiation therapy. CONCLUSION(S): Ovarian transposition is a safe and effective procedure for preserving ovarian function in patients treated by a radiosurgical combination. This procedure should be performed in patients <40 years of age with a small invasive cervical carcinoma (<3 cm) treated by initial surgery. In such selected cases, the risk of ovarian metastasis is low. PMID- 11020518 TI - Management of women with polycystic ovary syndrome who experienced premature luteinization during clomiphene citrate treatment. AB - OBJECTIVE: To determine the preferred treatment modality in patients with PCOS who experienced premature luteinization during CC treatment. DESIGN: Prospective randomized study. SETTING: Tertiary medical center. PATIENTS: Twenty-two infertile women with PCOS demonstrating premature luteinization during at least two consecutive CC cycles. INTERVENTIONS: Randomized induction of ovulation either with FSH alone or with GnRH agonist combined with FSH for a single treatment cycle. MAIN OUTCOME MEASURES: Premature luteinization was defined as serum progesterone >1.5 ng/mL before hCG administration. RESULTS: Premature luteinization occurred in eight of the 10 patients (80%) in group A and in two of the 12 patients in group B (16.6%). This result corresponds to the higher mean (+/-SD) progesterone level present in group A patients as compared to those in group B (2.0 +/- 1.2 ng/mL vs. 1.2 +/- 0.6 ng/mL, P=0.03). No pregnancies were achieved in group A, whereas the pregnancy rate per cycle observed in group B was 33.3% (4/12). On the day of hCG administration, the maximum mean (+/-SD) estradiol level was significantly lower (P<0.0001) in group A (210.6 +/- 37.9 pg/mL) than in group B (600.3 +/- 253.8 pg/mL). The treatment duration and the number of FSH ampules used did not differ between the groups. CONCLUSIONS: Pituitary desensitization with GnRH analog in combination with FSH is superior to FSH-only treatment in PCOS patients who demonstrate premature luteinization during CC treatment. PMID- 11020519 TI - Quantitative analysis of estrogen receptor alpha and beta messenger ribonucleic acid levels in normal endometrium and ovarian endometriotic cysts using a real time reverse transcription-polymerase chain reaction assay. AB - OBJECTIVE: To quantify messenger RNA (mRNA) levels of the two estrogen receptor isoforms, estrogen receptor-alpha (ER-alpha) and estrogen receptor-beta (ER-beta) in the eutopic endometrium and ovarian endometriotic cysts. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Patients with endometriosis and patients with uterine leiomyoma or carcinoma in situ. INTERVENTION(S): Gonadotropin-releasing hormone agonist (GnRH-a)-treated (n = 12) or untreated (n = 24) endometriotic cysts were obtained from 36 patients during laparoscopic cystectomy. Eutopic endometrium tissues were obtained from 24 patients during or immediately after surgery. MAIN OUTCOME MEASURE(S): ER-alpha and ER-beta mRNA levels, using a real-time reverse transcription (RT)-polymerase chain reaction (PCR) assay, TaqMan RT-PCR. RESULT(S): Eutopic endometrium and ovarian endometriotic cysts showed predominantly higher levels of ER-alpha mRNA than ER beta mRNA. Although ER-alpha and ER-beta mRNA levels in the eutopic endometrium were affected by a cyclic change in ovarian hormones, ovarian endometriotic cysts were less affected. Moreover, a long-term hypoestrogenic state induced by GnRH-a especially decreased ER-alpha mRNA levels in endometriotic cysts. Consequently, the relative ratios of ER-alpha to ER-beta mRNA levels in both GnRH-a-treated and untreated endometriotic cysts were significantly lower than those in the eutopic endometrium. CONCLUSION(S): The results suggest that the principal and regulatory effects of estrogens may be mediated mainly via ER-alpha rather than ER-beta in both the eutopic endometrium and endometriotic cysts. PMID- 11020520 TI - Apoptosis and expression of Bcl-2 and Bax in eutopic endometrium from women with endometriosis. AB - OBJECTIVE: To evaluate and compare spontaneous apoptosis and Bcl-2 and Bax expression in eutopic endometrium from women with and without endometriosis. DESIGN: Apoptosis and Bcl-2 and Bax expression were examined in eutopic endometrium from women with and without endometriosis. SETTING: Instituto de Biologia y Medicina Experimental-CONICET, Department of Gynecology and Department of Gynecological Pathology, Clinicas University Hospital, Buenos Aires, Argentina. PATIENT(S): Women with untreated endometriosis (n = 14) and controls (n = 16). INTERVENTION(S): Collection of endometrial samples during diagnostic or therapeutic laparoscopy. MAIN OUTCOME MEASURE(S): Apoptotic cells were detected with use of the dUTP nick-end labeling (TUNEL) assay; Bcl-2 and Bax expressions were assessed with use of immunohistochemical techniques. RESULT(S): Spontaneous apoptosis was significantly lower in eutopic endometrium from patients with endometriosis, compared with healthy controls (2.26 +/- 0.53 and 9.37 +/- 1.69 apoptotic cells/field, respectively) and was independent of cycle phase. An increased expression of Bcl-2 protein was found in proliferative eutopic endometrium from patients with endometriosis. Bax expression was absent in proliferative endometrium, whereas there was an increase in its expression in secretory endometrium from both patients and controls. CONCLUSION(S): Women with endometriosis show decreased number of apoptotic cells in eutopic endometrium. The abnormal survival of endometrial cells may result in their continuing growth into ectopic locations. PMID- 11020521 TI - Detection of endometrial pinopodes by light microscopy. AB - OBJECTIVE: To study the value of light microscopy (LM) in the assessment of endometrial pinopodes. DESIGN: Comparative histologic study. SETTING: Outpatient infertility clinic in an academic teaching institution. PATIENT(S): Eighteen oocyte donors undergoing controlled ovarian hyperstimulation. INTERVENTION(S): Endometrial biopsies on days 14-24 of the cycle. MAIN OUTCOME MEASURE(S): Assessment of pinopodes by scanning electron microscopy (SEM) and of endometrial surface projections by LM. RESULT(S): The luminal surface was identified by LM in 36 of 38 endometrial specimens obtained. Although apical projections could be recognized in all, they were few, moderate, and abundant in 20, 12, and 4 cases, respectively. Pinopodes were detected by SEM in all 4 samples with abundant projections, but in only 14 of 32 samples with lesser quantities of these surface features. No predictive value could be ascribed to apical projections viewed by LM for the developmental stage of pinopodes as defined by SEM. CONCLUSION(S): The LM of routine endometrial specimens can serve as a preliminary tool in the evaluation of surface morphology. Although abundant apical projections by LM are compatible with the presence of pinopodes by SEM, the latter modality remains as the definitive method in cases with few or moderate projections and for the evaluation of the stage of pinopode development. PMID- 11020522 TI - Do androgens have a direct effect on endometrial function? An in vitro study. AB - OBJECTIVE: To test the hypothesis that androgens have a direct effect on the function of endometrial epithelial cells. DESIGN: In vitro study. SETTING: Academic research center. PATIENT(S): Endometrial epithelial cells were prepared from biopsy samples obtained from normal fertile women. INTERVENTIONS: Cells were incubated with androstenedione, testosterone, dihydrotestosterone, and DHEA. MAIN OUTCOME MEASURE(S): Secretion of glycodelin A into the culture fluid was used to assess secretory activity. Uptake of (3)H-thymidine and immunostaining for Ki67 was used to assess cell growth. The specific action of the androgens was confirmed by incubation with an antiandrogen, cyproterone acetate. RESULT(S): Androstenedione (10(-6) M and 10(-7) M) caused a dose-dependent decrease in glycodelin A secretion, uptake of (3)H-thymidine, and percentage of positive Ki67 cells in cultured human endometrial epithelial cells. Testosterone, dihydrotestosterone, and DHEA had no effect on glycodelin A secretion or (3)H thymidine uptake. The direct effect of androgens on endometrial function were confirmed by demonstrating the presence of androgen receptors in cultured endometrial epithelial cells and showing that the direct effects of the androgens were not observed when cyproterone acetate was added to the cultures. CONCLUSION(S): The results suggest that androstenedione can inhibit human endometrial cell growth and secretory activity. Infertility and miscarriage associated with high androgen levels (e.g., that caused by the polycystic ovary syndrome) may be due to an adverse effect of high androgen levels on the endometrium. PMID- 11020523 TI - Importance of sperm-to-epithelial cell contact for the capacitation of human spermatozoa in fallopian tube epithelial cell cocultures. AB - OBJECTIVE: To investigate the mechanisms involved in the stimulatory effect of fallopian tube epithelial cell coculture on sperm movement characteristics. DESIGN: Human spermatozoa were cultured with human fallopian tube epithelial cell monolayers. A microporous membrane was used to prevent sperm-to-epithelial cell contact. Sperm movement characteristics were measured at 4 and 24 hours. SETTING: University hospital and fertility center. PATIENT(S): Voluntary donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Movement characteristics of human spermatozoa. RESULT(S): Fallopian tube epithelial cell coculture increased sperm motility, curvilinear velocity, amplitude of lateral head displacement, and hyperactivated motility, mainly at 24 hours, compared with controls. These stimulatory effects were inhibited when a microporous membrane prevented cell-to cell contact between sperm and fallopian tube epithelial cells. CONCLUSION(S): Physical contact between sperm and epithelial cells in coculture systems seems to be the main factor in stimulating sperm movement characteristics, and this could be the main mechanism of in vivo sperm capacitation. PMID- 11020524 TI - Identification of meiotic and postmeiotic gene expression in testicular tissue of patients histologically classified as Sertoli cell only. AB - OBJECTIVE: To determine whether meiotic and postmeiotic germ cell gene products could be detected in biopsy specimen from patients with Sertoli cell only (SCO) and maturation arrest. DESIGN: Prospective clinical study. SETTING: University based departments and laboratories. PATIENT(S): Nine patients, seven with nonobstructive azoospermia (12 biopsies) and two with obstructive azoospermia (controls) (2 biopsies). INTERVENTION(S): Specimens were divided into three parts: IVF laboratory, histology, and molecular analysis. Germ cell-specific messenger RNAs (mRNAs) were detected by extracting total RNA for Northern blotting or reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURE(S): Detection of meiotic (lactate dehydrogenase C4) and postmeiotic (transition protein 1 and protamine 1 and 2) gene expression and correlation with histologic and IVF laboratory findings. RESULT(S): The IVF laboratory identified spermatozoa in 3 of 14 biopsies (controls and severe hypospermatogenesis). Histologically, 6 of 14 biopsies (43%) were diagnosed as SCO, 4 (29%) maturation arrest, 2 (14%) severe hypospermatogenesis, and 2 normal. Molecular analysis showed mRNA for meiotic and postmeiotic genes in 12 of 14 biopsies (86%) (P =. 006), of which 4 (67%) in SCO and 3 (75%) in maturation arrest. CONCLUSION(S): Differentiated germ cells are present in biopsies of men histologically diagnosed as SCO. Absence of these molecular markers strengthens the histologic diagnosis and helps the physician in counseling the infertile couple. PMID- 11020525 TI - Endometrial laser intrauterine thermotherapy: the first series of 100 patients observed for 1 year. AB - OBJECTIVE: To describe a new instrument (GyneLase) that offers a new approach (endometrial laser intrauterine thermal therapy [ELITT]) to treatment of menorrhagia and to evaluate the efficacy of ELITT in the management of dysfunctional uterine bleeding. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): 100 premenopausal women with dysfunctional uterine bleeding were observed for 1 year. INTERVENTION(S): Intrauterine laser thermotherapy with a diode laser. MAIN OUTCOME MEASUREMENT(S): Amenorrhea rate after 1 year. RESULT(S): The amenorrhea rate after 1 year of follow-up was 71%, and the rate of amenorrhea/severe hypomenorrhea rate was >90%; these rates are much higher than those in the literature after such procedures as electrosurgery or intrauterine thermal balloon therapy. The ELITT procedure is an inherently safe and simple alternative, providing controlled and effective treatment of the entire endometrium. In contrast to traditional endometrial ablation using a neodymium yttrium-aluminum-garnet laser, the ELITT procedure does not require intensive training or hysteroscopic control; it is also far less risky, because the power used per unit area is 1,000 times lower. CONCLUSION(S): The ELITT procedure is a new nonhysteroscopic technique for endometrial ablation. The technique is very safe and offers the highest amenorrhea rate to date in the literature. PMID- 11020526 TI - Role of three-dimensional ultrasonographic measurement of endometrium volume as a predictor of pregnancy outcome in an IVF-ET program: a preliminary study. AB - OBJECTIVE: To evaluate the role of three-dimensional (3D) ultrasonographic measurement of the endometrium in predicting pregnancy in an IVF-ET program. DESIGN: Prospective study. SETTING: Center for assisted reproductive techniques. PATIENT(S): 65 women undergoing controlled ovarian hyperstimulation for IVF. INTERVENTION(S): Ultrasonographic examination on the day of hCG administration. MAIN OUTCOME MEASURE(S): Endometrial volume, endometrial thickness, and pregnancy rate. RESULT(S): The mean (+/-SD) endometrium volume (4.16 +/- 1.97 mL), endometrium thickness (11 +/- 2 mm), and estradiol level (1686.82 +/- 1057.10 pg/mL) in 21 pregnant women on the day of hCG administration did not differ statistically differ from the respective values in 44 nonpregnant women (4.53 +/- 1.79 mL; 11 +/- 2 mm, 1883.56 +/- 1147.21 pg/mL). Receiver-operating characteristic curves showed that the area under curve (AUC) was 0.57 for endometrial volume and 0.48 for endometrial thickness. Using a cut-off value of 2.5 mL for endometrial volume to predict pregnancy, results of the Fisher exact test were statistically significant. No significant cut-off value was found for endometrial thickness. CONCLUSION(S): Endometrial volume and thickness on the day of hCG administration did not predict occurrence of pregnancy. A minimum volume of 2.5 mL appeared to favor pregnancy. The predictive value of 3D ultrasonographic measurement of endometrial volume and thickness was better than that of 2D measurement. PMID- 11020527 TI - Cryopreservation of mature human oocytes by vitrification with ethylene glycol in straws. AB - OBJECTIVE: To examine the effect of vitrification with ethylene glycol (EG) for mature human oocytes in straws. DESIGN: Prospective, randomized, in vitro experiments. SETTING: Reproductive unit of a university hospital. PATIENT(S): Immature oocytes from 110 patients undergoing intracytoplasmic sperm injection (ICSI). INTERVENTION(S): The immature oocytes were incubated to reach metaphase II (MII). The MII oocytes were treated with EG-based cryoprotectants and vitrified in straws. They were diluted in sucrose solutions, inseminated by ICSI, and cultured in vitro. MAIN OUTCOME MEASURE(S): Survival, fertilization, and embryo cleavage. RESULT(S): The survival rates were greater for oocytes pretreated with 1.5 M of EG (65% for 0 minute, 93% for 5 minutes, and 96% for 10 minutes). The oocytes vitrified in 60 and 90 seconds had a greater rate of fertilization than those vitrified in 120 seconds. There were no differences in survival and fertilization for vitrified oocytes diluted by three or four steps. The cleavage rates to the six- to eight-cell stage were comparable with controls. However, no blastocyst formation was observed in vitrified oocytes. CONCLUSION(S): Vitrification of human oocytes with EG in straws achieves a high rate of survival, fertilization, and early cleavage of embryos. Further studies should be conducted for the improvement of blastocyst formation. PMID- 11020528 TI - Perifollicular Doppler flow and follicular fluid vascular endothelial growth factor concentrations in poor responders. AB - OBJECTIVE: To prospectively evaluate follicular fluid levels of vascular endothelial growth factor in women undergoing IVF cycles and to investigate the correlation of these levels with ovarian response to gonadotropins and with uterine or ovarian Doppler findings. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): 41 patients undergoing ART were divided into two groups according to response to ovarian stimulation protocols: poor responders (n = 18) and normoresponders (n = 23). INTERVENTION(S): Doppler analysis of perifollicular arteries and assay of follicular fluid vascular endothelial growth factor. MAIN OUTCOME MEASURE(S): During ovarian stimulation, patients underwent hormonal (E2), ultrasonographic (follicular number and diameter, endometrial thickness) and Doppler (uterine and perifollicular arteries) evaluation. Serum and follicular fluid concentrations of vascular endothelial growth factor were assayed in each female patient. RESULT(S): Compared with poor responders, more oocytes were collected and more embryos were transferred but follicular fluid levels of vascular endothelial growth factor levels were lower in normoresponders. Follicular fluid levels of vascular endothelial growth factor were inversely correlated with number of oocytes retrieved. Poor responders had significantly higher uterine and perifollicular Doppler flow resistances. The pregnancy rate per cycle was significantly higher in normoresponders (26%) than poor responders (6%). CONCLUSION(S): Elevated follicular fluid levels of vascular endothelial growth factor concentrations are associated with poor ovarian response and a very low pregnancy rate. PMID- 11020529 TI - Impact of implementation of an embryo storage fee on embryo disposal activity. AB - OBJECTIVE: To examine the impact of implementation of a new fee for continued storage of cryopreserved embryos on the rate of requests for disposal of embryos. DESIGN: Retrospective cohort study. SETTING: A university-based assisted reproduction program. PATIENT(S): All patients with cryopreserved embryos. INTERVENTION(S): Implementation of a semiannual embryo storage fee of $100 to cover administrative and laboratory costs. MAIN OUTCOME MEASURE(S): The number of embryo disposal requests before and after implementation of the embryo storage fee was compared in relation to the activity of the cryopreserved embryo program as measured by number of frozen embryo transfers. RESULT(S): Annual requests for embryo disposal from 1992 through 1997 ranged from zero to three, which represented 0-5% of the annual frozen embryo program activity. In contrast, a significantly higher number of disposal requests (10, representing 18% of program activity) were received in 1998. CONCLUSION(S): Fees for storage of cryopreserved embryos seem to influence patients' decisions about disposal of cryopreserved embryos. PMID- 11020531 TI - Impact of reducing the number of embryos transferred from three to two in women under the age of 35 who produced three or more high-quality embryos. AB - OBJECTIVE: To investigate the effect of a reduction in the number of good-quality embryos transferred in patients <35 years of age on pregnancy and multiple pregnancy rate. DESIGN: Prospective observational study with historical controls. SETTING: Academic tertiary referral unit. PATIENT(S): Three hundred eight patients <35 years of age undergoing IVF-ET. INTERVENTION(S): For patients who had three or more good quality embryos available for transfer, those in group 1 were given the option to have either two or three embryos replaced, whereas those in group 2 were allowed a maximum of two embryos transferred. In both groups, patients who had less than three good-quality embryos had the option to have three embryos transferred. MAIN OUTCOME MEASURE(S): Pregnancy and multiple pregnancy rates. RESULT(S): Patients in group 1, compared with those in group 2, had significantly more embryos (3 vs. 2) of significantly higher cumulative embryo score (31 vs. 24) transferred. This resulted in significantly higher multiple (57.8% vs. 30.8%) and triplet (15.6% vs. 1.4%) pregnancy rates in group 1. However, no difference in overall clinical pregnancy rate (37.2% vs. 41.2%) or live birth rate (28.1% vs. 29.4%) was observed between group 1 and 2. CONCLUSION(S): In women <35 years of age, who have three or more good-quality embryos available for transfer, a maximum of two embryos should generally be transferred. PMID- 11020530 TI - Treatment of hirsutism with ethinyl estradiol-desogestrel contraceptive pills has beneficial effects on the lipid profile and improves insulin sensitivity. AB - OBJECTIVE: To evaluate the effects on the lipid pattern and insulin sensitivity of hirsute women of an oral contraceptive pill containing 30 microg of ethinyl estradiol and 150 microg of desogestrel. DESIGN: Prospective clinical study. SETTING: Tertiary care institutional hospital. PATIENT(S): 16 hirsute women. INTERVENTION(S): Women were evaluated at baseline and after receiving six cycles of oral contraceptive therapy. MAIN OUTCOME MEASURE(S): Body mass index (BMI); hirsutism score (nine body areas); serum levels of total cholesterol, high density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, triglycerides, apolipoprotein B, lipoprotein(a), and serum adrenal and ovarian androgens; and fasting glucose and insulin concentrations. RESULT(S): The mean serum total, HDL, and LDL cholesterol levels increased after six cycles of oral contraceptive therapy. Levels of HDL cholesterol were < 50 mg/dL in 7 of the 16 patients at baseline; these levels normalized in 4 patients after treatment. Serum total and LDL cholesterol remained within the normal range in all patients before and after therapy. No significant changes were observed in serum triglyceride, apolipoprotein B and lipoprotein(a) concentrations. Fasting insulin levels and insulin resistance as analyzed by homeostasis model assessment were reduced significantly after therapy. No changes in BMI were observed. Administration of oral contraceptive pills signifiCantly reduced the hirsutism score and hyperandrogenemia. CONCLUSION(S): Oral contraceptive pills containing low-dose ethinyl estradiol and desogestrel are effective in controlling hyperandrogenism and hirsutism and ameliorate the abnormal metabolic profile of women with hirsutism. PMID- 11020532 TI - Influence of initial semen quality on the integrity of human sperm DNA following semen processing. AB - OBJECTIVE: To examine and compare the effects of density-gradient centrifugation on the integrity of sperm DNA from the semen of both fertile and infertile men. DESIGN: Prospective, observational study. SETTING: University infertility clinic. PATIENTS: Forty-four nonazoospermic, infertile men and nine fertile controls. INTERVENTIONS: Semen samples were processed by density-gradient centrifugation. Sperm motility and sperm chromatin structure (evaluated by flow cytometry analysis of acridine orange-treated spermatozoa) were monitored before and after semen was processed. MAIN OUTCOME MEASURES: Sperm motility and DNA integrity. RESULTS: Following density-gradient centrifugation, mean sperm motility (+/-SEM) improved significantly compared to whole semen in samples from fertile and infertile men, respectively (71 +/- 6 vs. 49 +/- 7% and 56 +/- 3 vs. 44 +/- 3%, P<0.05). However, the percentage of sperm with denatured DNA increased compared to whole semen after processing of samples from infertile (25 +/- 3 vs. 15 +/- 2%, P<0. 01) but not fertile men (9 +/- 3 vs. 8 +/- 2%, P>0.05). CONCLUSIONS: Our data indicate that the potential detrimental effect of density-gradient centrifugation on sperm DNA integrity is related to the initial semen quality. These data urge us to examine our current sperm-processing techniques to minimize sperm DNA damage. PMID- 11020533 TI - Partial hydatidiform mole following transfer of a cryopreserved-thawed blastocyst. PMID- 11020534 TI - Hyperimmunoglobulin therapy for cytomegalovirus-associated infertility: case reports. PMID- 11020535 TI - Birth of healthy male twins after intracytoplasmic sperm injection of frozen thawed testicular spermatozoa from a patient with nonmosaic Klinefelter syndrome. PMID- 11020536 TI - Use of the Multi-ZSC one-step standardized swim-up method: recovery of high quality spermatozoa for intrauterine insemination or other forms of assisted reproductive technologies. PMID- 11020537 TI - Pregnancy and delivery of healthy infants developed from vitrified blastocysts in an IVF-ET program. PMID- 11020538 TI - Research contributions to Fertility and sterility in the field of gynecology at the change of the millennium. PMID- 11020540 TI - Letter to the editor PMID- 11020539 TI - Effect of injected spermatozoa morphology on the outcome of intracytoplasmic sperm injection in humans. PMID- 11020541 TI - Errata PMID- 11020542 TI - Consultation-liaison psychiatry training and supervision results in fewer recommendations for constant observation. AB - In this study, we tested two hypotheses. First, that consultation-liaison (C-L) trained psychiatry residents would order constant observation (CO) less frequently than psychiatry residents untrained in C-L. Second, we predicted that CO would be ordered less frequently under circumstances when experienced C-L psychiatry attending and fellows would be available to supervise psychiatry residents training in C-L. We reviewed a total of 138 consultations during a 6 month period. Constant observation was recommended in 31 cases (22.5%). Consultations were done by residents who had received training in C-L psychiatry (n=34) and by residents who were not trained in C-L (n=34). Residents not trained in C-L had a significantly higher percentage of CO orders (44.1%) compared to those trained in C-L (15.4%) (chi(2)=12.1, df=1, P<0.001). Because C-L-trained residents provided regular-hour and after-hour consults while residents without C L training provided only after-hour consults, we also separately analyzed data from the 102 after-hour cases. We again found that residents with C-L training had a significantly lower rate of ordering CO (22.1%) than those who had not yet received C-L psychiatry training (44.1%) (chi(2)=5.31, df=1, P<0.05). We also found that C-L-trained residents ordered CO less frequently during regular hour consults (2.8%) when experienced staff are available in supervision compared to after hours (22.1%) (chi(2)=6.72, df=1, P<0.01). Our findings suggest that training in C-L psychiatry has a significant impact on the use of constant observation for patients in the general hospital thereby reducing the cost of care. PMID- 11020543 TI - Depression with irritability in patients undergoing coronary artery bypass graft surgery: the cardiologist's role. AB - The purpose of this study was to investigate the determinants and maintenance factors of depression in patients undergoing coronary artery bypass graft (CABG) surgery. Fifty CABG candidates were selected randomly for evaluation before and after surgery. Depressive disorders occurred in 22.0% of the patients preoperatively and 21% postoperatively. Irritability as a symptom was more prevalent in depressed than nondepressed patients preoperatively (P=.04) and postoperatively (P=.006). Although depression was the most prevalent diagnosis in the patients' psychiatric histories and evaluations, benzodiazepines were prescribed more frequently than antidepressants (70% vs 3%). Cardiologists should consider that the irritability in some patients undergoing CABG may obscure the diagnosis of depression. The correct recognition of depression is essential to reduce inappropriate benzodiazepine prescription and to apply appropriate treatment. PMID- 11020544 TI - Maximizing life's potentials in AIDS: a psychopharmacologic update. AB - Psychiatric factors play a major role in the AIDS pandemic. They have an impact on transmission, morbidity, coping, adherence, and quality of life and of death. Substance-related disorders are associated with HIV transmission through needle sharing, sexual transmission, exchange of sex for drugs, and perinatal transmission. Persons with AIDS have a high prevalence of substance-related disorders, mood disorders, dementia, mania, and delirium. Persons with AIDS require complex medications, including combination antiretroviral therapy and prophylaxis and treatment for opportunistic infections and cancers. Recognition and treatment of distressing symptoms can maximize life's potentials and enhance adherence with risk prevention and with care. We describe an integrated approach to the use of psychotropic medications in the care of persons with AIDS. With the new antiretroviral therapies available, psychotropic medications can be helpful in alleviating distressing symptoms, promoting less risk taking, and adhering to complex medical care. The intricacies and complexities of new medical and psychopharmacological issues are delineated in order to enable caregivers help persons with AIDS to maximize life's potentials. PMID- 11020545 TI - Exogenous BDNF, NT-3 and NT-4 differentially regulate neurite outgrowth in cultured hippocampal neurons. AB - Multiple growth factors contribute to the differentiation of dendritic and axonal processes by a neuron. Cultured hippocampal cells elaborate dendritic and axonal processes following well-defined steps. We used this culture system to determine the specific effects of brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4) on dendritic and axonal differentiation in hippocampal pyramidal neurons. We demonstrated that each of these neurotrophins exert distinct effects on neurite outgrowth. Both BDNF and NT-3 had positive effects on the outgrowth of undifferentiated neurites, called minor neurites, and on the axonal process of hippocampal pyramidal neurons. However, the effect of NT 3 was more important than that of BDNF. On the other hand, NT-4 did not enhance axonal outgrowth but had only an effect on the outgrowth of minor neurites. Since cytoskeletal proteins play crucial roles in promoting neurite outgrowth, we examined the protein levels of some of these proteins that are associated with neurite outgrowth: beta-actin, gamma-actin, alpha-tubulin, MAP2 and tau. Surprisingly, we did not detect any change in their protein levels. Taken together, our results show that BDNF, NT-3 and NT-4 exert distinct effects on the neuritic compartments of hippocampal neurons. PMID- 11020546 TI - Lithium treatment in ovo: effects on embryonic heart rate, natural death of ciliary ganglion neurons, and brain expression of a highly conserved chicken homolog of human MTG8/ETO. AB - Understanding the action of the mood stabilizer lithium is dependent on availability of experimental models where lithium treatment at clinically relevant concentrations induces marked phenotypic and genotypic changes. Here we report on such changes in the chicken embryo. Lithium chloride (0.6 mM), applied in ovo 60 h after incubation, markedly delayed the heart rate increase observed from ED2.5 to ED5, and induced the brain expression of a new chicken gene cETO from ED7 to ED15. At the same time the overall developmental dynamics and embryo survival, or the expression of chicken gephyrin were not significantly affected. Furthermore, lithium treatment (0.3 mM, 48 h after incubation) abolished the difference in neuronal number between ED12 ciliary ganglia developing in the presence or absence of postganglionic target muscles. We show that cETO is a close homologue of the human transcription factor MTG8/ETO; named after its location on chromosome 8, and participation in chromosomal translocation 8;21 in myeloid leukemia. The mRNA and protein levels of ETO and gephyrin had a parallel course in chicken brain development suggesting that the expression of both genes is regulated mainly at the level of gene transcription. However, the patterns of expression were markedly different. ETO peaked at ED7 and decreased five-fold at ED15. In contrast, gephyrin levels increased five-fold from ED7 to ED15. We propose that the induction of ETO expression, in concert with lithium-induced upregulation of other genes, such as PEBP2beta and bcl-2, is participating in the neuroprotective effect of chronic lithium treatment. PMID- 11020547 TI - Prenatal choline exposure alters hippocampal responsiveness to cholinergic stimulation in adulthood. AB - Manipulation of dietary choline levels during gestation results in enduring neurobehavioral changes in offspring that last into adulthood. Alterations of hippocampal function and memory are among the most striking changes. Depending upon the measures assessed, prenatal choline supplementation tends to promote excitatory synaptic efficacy in hippocampal circuits while prenatal choline deficiency diminishes it. However, the mechanisms underlying these changes remain unclear. Transverse hippocampal slices were prepared from adult offspring of dams fed choline supplemented, choline deficient, or control diets. We assessed paired pulse inhibition, and excitatory synaptic responsiveness before and after activation of cholinergic receptors with Carbachol. Prenatally choline deficient animals yielded significantly fewer electrophysiological viable hippocampal slices than did animals from either of the other two treatment groups. Among the slices tested, there were no differences in paired pulse inhibition between the treatment groups. However, transient cholinergic activation resulted in a prolonged enhancement of the amplitude of the population EPSP (pEPSP) response in slices from prenatally choline supplemented animals. These results suggest that GABA receptor-mediated inhibition remains intact after prenatal choline manipulations, and that enhancement of the excitatory responsiveness of hippocampal circuits in slices from prenatally choline supplemented rats may be related in part to an increase in cholinergic tone within the CA1 circuit. PMID- 11020548 TI - Proliferation and migration of receptor neurons in the vomeronasal organ of the adult mouse. AB - Cell proliferation and differentiation in the vomeronasal organ of the adult mouse was studied by bromodeoxyuridine (BrdU) immunohistochemistry coupled to immunostaining for specific markers of the differentiation, such as carnosine, B50-GAP43 (growth-associated protein) and stathmin. The present study shows that three populations of proliferating elements are present in the vomeronasal sensory epithelium that are placed, respectively, in the supporting cell layer, at the boundaries between the sensory epithelium (S-VNO) and the non-sensory (NS VNO) and in the basal region of the S-VNO. The number of dividing cells at the boundaries of the S-VNO is by far prevailing. Few proliferating cells located adjacent to the basal membrane are, however, present 1 day after BrdU inoculations. Seven days after BrdU treatment immunopositive nuclei were detected in more central regions of the VNO and at longer survival times they were also positive to carnosine, a marker of fully differentiated neurons. In conclusion, the present results suggest that at least two populations of VNO neuronal precursors are responsible for cell replacement throughout life. PMID- 11020549 TI - Postnatal developmental changes in AMPA and NMDA receptors in the rat vestibular nuclei. AB - Changes in the expression of the AMPA receptor subunits GluR1-4 and of the NMDA receptor subunits NR1, NR2A-D were investigated in the developing rat medial and lateral vestibular nuclei. Analyses were performed using nonradioactive in situ hybridization and immunoblotting with subunit-specific antibodies. During the postnatal development, glutamatergic receptor subunits were differentially expressed in the vestibular nuclei. The level of expression of GluR1, GluR4 and NR1 subunits was higher in the developing brain as compared to the adult. We observed a gradual increase in GluR2/3, NR2A, NR2B and NR2C levels of expression in the medial and lateral vestibular nuclei during the first 3 weeks of postnatal development. In situ hybridization results were consistent with immunoblot analyses. The differential expression of AMPA and NMDA receptor subunits in immature vestibular neurons is consistent with changes in glutamate receptor properties. This may be related to the postsynaptic regulation of receptor subunits associated with the synaptic plasticity of the vestibular neuron connections during specific sequences of postnatal development. PMID- 11020550 TI - Perinatal distribution of galanin and galanin receptor-1 mRNA in the rat hindbrain. AB - In situ hybridisation was used to determine the distribution and levels of mRNA for galanin precursor preprogalanin (ppGAL) and galanin receptor-1 (GAL-R1) in the rat hindbrain before and after birth. Quantification of mRNA levels was performed from E21. Also, immunohistochemistry was used to study GAL-like immunoreactivity (GAL-LI) prenatally. On E16, no expression of ppGAL mRNA could be detected in any areas examined, whereas on E19 low transcript levels were observed. GAL-LI, however, was seen at relatively high levels in nerve fibres already on E16, mainly in the areas receiving primary afferents. Also, GAL-R1 mRNA was expressed at high levels in discrete areas of the hindbrain on E16. On E21 ppGAL mRNA was found in the locus coeruleus (LC), the nucleus of the solitary tract, the dorsal motor nucleus of the vagus (nX), the lateral reticular nucleus (LRn) and superficially along the ventral medullary surface. Expression increased postnatally in all these areas except in nX and LRn. GAL-R1 mRNA, on the other hand, was found to be expressed at high levels on E21 in the LC, where levels then decreased on P1. Expression of GAL-R1 mRNA was also found in other areas of the brainstem, but here no changes were detected around birth. These findings demonstrate that ppGAL and GAL-R1 mRNAs, as well as GAL-LI, are present in the brainstem in the rat fetus and that the changes in expression after birth could be of importance for the newborn in the transition from pre- to postnatal life. PMID- 11020551 TI - Postnatal development of somatostatin 2A (sst2A) receptors expression in the rabbit retina. AB - In the retina, somatostatin (SRIF) acts as a neuromodulator by interacting with specific SRIF subtype (sst) receptors. Aim of this investigation was to determine the cellular localization of the sst2A receptor isoform in the postnatal rabbit retina. Receptor immunoreactivity was localized using the antiserum K-230, directed to the C-terminus of the human sst2A receptor. In the postnatal rabbit retina, sst2A receptors were abundantly expressed without significant regional differences. They were localized predominantly to rod bipolar cells, identified with a protein kinase C (PKC) antibody, to amacrine cells, some of which also containing tyrosine hydroxylase (TH), and to presumed rare horizontal cells. Quantitative analysis showed that sst2A-immunoreactive (-IR) bipolar and amacrine cells reached their maximum density and absolute number at the time of eye opening, when the expression pattern of sst2A receptors was similar to that in adult retinas. In the adult retina, 68% of the PKC-IR rod bipolars and 34% of the TH-IR amacrine cells were observed to also express sst2A receptors. The appearance of sst2A receptor immunolabeling prior to eye opening and the developmental profile of sst2A receptor expression are compatible with a role of SRIF in the maturation of retinal circuitries. The partial expression of sst2A receptors in PKC-IR rod bipolar cells and in TH-IR amacrine cells may suggest some type of heterogeneity within these cell populations. PMID- 11020552 TI - Localization of p450scc and 5alpha-reductase type-2 in the cerebellum of fetal and newborn sheep. AB - Prenatally, neuroactive steroids that modulate GABAergic activity may be synthesized de novo within the fetal brain. We have examined changes in immunoreactivity staining for the steroidogenic enzymes cholesterol P450 side chain cleavage (P450scc), and 5alpha-reductase type-2 in the cerebellum of late gestation (130-145 days gestation) fetal sheep and newborn lambs (1-4 weeks of age). Both enzymes were predominantly localized in the Purkinje cell body and dendrites of the fetal and newborn cerebellum, with weaker immunoreactivity in a few cells of the inner granular layer. P450scc immunoreactivity was present in Purkinje neurons expressing either of the neuronal microtubule associated proteins MAP1b/5 or MAP2a/b, but was absent from GFAP and HNK-1 positive cells. Soma of Purkinje neurons were also immunopositive for 5alpha-reductase type-2 in the fetuses, but expression decreased to just detectable levels in the 1-2 and 2 4 week old lambs. Both MAP1b/5- and MAP2a/b-positive Purkinje neurons showed 5alpha-reductase type-2 expression in the fetus, whereas the residual 5alpha reductase staining in the newborn lamb was present only in MAP2a/b-positive Purkinje neurons. Allopregnanolone in the cerebellum decreased from 21.8+/-1.9 ng/g wet weight in fetuses at 140-145 days gestation to 6.7+/-0.5 ng/g in 2-4 week old lambs (P<0. 05). Thus, synthesis of neuroactive steroids from cholesterol is possible in cerebellar neurons in late gestation and persists into neonatal life, 5alpha-reductase type-2 expression is greater in the fetus compared to the neonate, and allopregnanolone concentrations in the cerebellum decrease significantly after birth. PMID- 11020553 TI - Neurotrophin-4/5 promotes proliferation of oligodendrocyte-type-2 astrocytes (O 2A). AB - In this study, the ability of neurotrophin-5 (NT-4/5) to promote the proliferation of oligodendrocyte precursor (O-2A) cells has been examined. This has been accomplished by the addition of exogenous NT-4/5 to purified cultures of O-2A cells maintained in an undifferentiated state by the addition of the mitogens platelet derived growth factor (PDGF-AA) and basic fibroblast growth factor (bFGF). Counts of cells incorporating bromodeoxyuridine (BrdU) showed that the addition of NT-4/5 for 24 h increased O-2A cell proliferation by 1.8-fold above that seen in PDGF-AA and bFGF alone. These data demonstrate a previously unidentified role for the neurotrophin NT-4/5 in oligodendrocyte biology. PMID- 11020554 TI - Prenatal methotrexate exposure delays onset of low Mg(2+)-induced epileptiform discharges in the entorhinal cortex. AB - We determined the effects of prenatal exposure to DNA synthesis inhibitor methotrexate (MTX) on: (a) the susceptibility to low Mg(2+)-induced epileptiform activity in deep layers (IV-V) of medial entorhinal cortex in vitro; and (b) neuronal counts in this area. Low Mg(2+)-induced discharges developed significantly later in slices from prenatally MTX-exposed rats than in control slices. Neuronal counts were increased in the layer V of medial entorhinal cortex of prenatally MTX-exposed rats. Results indicate that: (a) MTX-induced prenatal brain DNA impairment is antiepileptogenic; and (b) simple increases in neuronal numbers may not be associated with epileptogenic effects. PMID- 11020555 TI - Immunogold cytochemistry of the blood-brain barrier glucose transporter GLUT1 and endogenous albumin in the developing human brain. AB - The blood-brain barrier (BBB) glucose transporter, GLUT1, was detected by immunogold electron microscopy on the microvascular compartment of the human foetus telencephalon at the 12th and 18th weeks of gestation. By computerized morphometry, the cellular and subcellular localization of the immunosignal for GLUT1 was quantitatively evaluated. The study showed that the glucose transporter is strongly expressed by endothelial cells while a very low signal is detected on vascular pericytes. The GLUT1 antigenic sites are preferentially associated to the ablumenal and junctional plasma membranes of the endothelial cells and tend to increase significantly with age. A parallel study carried out by the endogenous serum protein albumin demonstrated that already at the 12th week the endothelial routes are hindered to the protein as happens at the blood endothelium interface of mature brain. The results demonstrate that in the human foetus the brain microvessels express BBB-specific functional activities early. PMID- 11020556 TI - Postoperative radiotherapy for lung cancer: the breast cancer story all over again? PMID- 11020557 TI - Overall survival after prostate-specific-antigen-detected recurrence following conformal radiation therapy. AB - PURPOSE: To study the significance, in terms of overall and cause-specific survival, of biochemical failure after conformal external-beam radiation therapy (RT) for prostate cancer. METHODS AND MATERIALS: Of the 1844 patients in the Radiation Oncology prostate cancer database, 718 were deemed eligible. Patients excluded were those with N1 or M1 disease, those treated after radical prostatectomy, those who received hormone therapy before radiation therapy, and those who died, failed clinically, or had no PSA response in the first 6 months after RT. Patients included were required to have a minimum of 2 post-RT PSAs separated by at least 1 week. Biochemical relapse was defined as 3 consecutive PSA rises. This resulted in 154 patients with biochemical failure. Survival was calculated from the third PSA elevation. The rate of rise of PSA was calculated by fitting a regression line to the four rising PSAs on a ln PSA vs. time plot. RESULTS: There were 41 deaths among the 154 patients with failure in 23 of the 41 due to prostate cancer. The overall survival after failure was 58% at 5 years, while the cause-specific failure was 73% at 5 years. Among the 154 failures, several factors were evaluated for an association with overall survival: age at failure, pre-RT PSA, PSA at second rise, PSA nadir, time from RT to failure, time to nadir, Gleason score, T-stage, and rate of rise, both from the nadir and from the beginning of the rise. None of these factors were significantly associated with an increased risk of death. As expected, the group of patients with biochemical failure have significantly worse prognostic factors than those without biochemical failure: median pre-RT PSA 15.9 vs. 9.0 (p < 0.001), and Gleason score of 7 or greater for 48% of subjects vs. 40% (p = 0.1). Relative PSA rise and slope of ln PSA vs. time were associated with cause-specific mortality (p < 0.001 and p = 0.007, respectively). CONCLUSION: Overall survival after conformal radiotherapy for prostate cancer remains high 5 years after biochemical failure. This high survival rate occurs even though the group of patients with biochemical failure has worse than average adverse preradiation prognostic factors. Thus, although biochemical failure can identify patients who have recurrent disease after RT, the ultimate relationship between this endpoint and death remains to be better defined. PMID- 11020558 TI - Complications from radiotherapy dose escalation in prostate cancer: preliminary results of a randomized trial. AB - OBJECTIVE: To compare early and late side effects in prostate cancer patients with Stage T1b-T3 disease randomized to receive 70 Gy or 78 Gy. METHODS: There were 189 patients randomized with a minimum follow-up of 2 years, that were available for this analysis. All patients were initially treated with a 4-field box to an isocenter dose of 46 Gy at 2 Gy per fraction. In the 70-Gy arm, treatment was continued to a reduced volume using a 4-field box technique. In the 78-Gy arm, treatment was continued to a reduced volume using a conformal 6-field arrangement. Side effects were graded on a 1-4 scale, adapted from Radiation Therapy Oncology Group and Late Effects Normal Tissue Task Force criteria. RESULTS: No significant differences in acute rectal or bladder toxicity were seen between the two treatment techniques (p > 0.6 for all comparisons). The 5-year Kaplan-Meier risks of Grade 2 or higher late bladder toxicity were 20% and 9% for 70-Gy and 78-Gy groups, respectively (log rank, p = 0.8). The 5-year risks of Grade 2 or higher late rectal toxicity were 14% and 21% for 70 Gy and 78 Gy, respectively (p = 0.4). Dose-volume histogram analysis of the 78-Gy patients showed a significant correlation between the percentage of rectum irradiated to 70 Gy or greater and the likelihood of developing late rectal complications. Patients with more than 25% of the rectum receiving 70 Gy or greater had a 5-year risk of Grade 2 or higher complications of 37% compared to 13% for patients with 25% or less (p = 0.05). All three Grade 3 complications occurred when greater than 30% of the rectum received 70 Gy or more. CONCLUSION: The overall rate of complications was similar in both treatment arms. However, there is evidence for a significant increase in late rectal complications when more than 25% of the rectum received 70 Gy or greater. This parameter may serve as a benchmark for the design of future three-dimensional conformal trials. PMID- 11020559 TI - Long-term urinary toxicity after 3-dimensional conformal radiotherapy for prostate cancer in patients with prior history of transurethral resection. AB - PURPOSE: To report on the long-term urinary morbidity among prostate cancer patients with a prior history of a transurethral resection of the prostate (TURP) treated with high-dose 3-dimensional conformal radiotherapy (3D-CRT). METHODS AND MATERIALS: Between 1988 and 1997, 1100 patients with clinically localized prostate cancer were treated with 3D-CRT. Of these, 120 patients (8%) were identified as having had a prior TURP and are the subjects of this analysis. The median age was 71 years (range: 49-83 years). The clinical stages of the patients were T1c: 33 (28%); T2a: 38 (32%); T2b: 15 (13%); and T3: 34 (27%). Neoadjuvant androgen ablation therapy was given to 39 (33%). The median radiation dose prescribed to the planning target volume was 75.6 Gy (range: 64.8-81 Gy). The median elapsed time from TURP to initiation of 3D-CRT was 69 months (range: 4-360 months). The median follow-up time was 51 months (range: 18-109 months). RESULTS: Five patients of the 120 with a prior history of TURP (4%) developed a urethral stricture after 3D-CRT which was corrected with dilatation. The 5-year actuarial likelihood of >/= Grade 2 late urinary toxicities was 9%. No Grade 4 urinary toxicities were observed in this group of patients. Among 110 patients who were completely continent of urine prior to 3D-CRT, 10 (9%) developed stress incontinence requiring 1 pad daily for protection or experienced occasional leakage (not requiring pad protection). The 5-year incidence of >/= Grade 1 stress incontinence was 18% in patients who developed acute >/= Grade 2 GU symptoms during the course of 3D-CRT compared to 7% for patients who experienced Grade 1 or no acute urinary symptoms (p = 0.05). The radiation dose (>/=75.6 Gy vs. <75.6 Gy), the number of prior TURP procedures, or the volume of resected tissue at the time of TURP had no significant impact on the long-term urinary morbidity outcome. A multivariate analysis demonstrated that the presence of Grade 2 acute urinary symptoms was the only predictor of >/= Grade 1 stress incontinence after 3D-CRT in this group of patients. CONCLUSIONS: Despite prior TURP, the incidence of >/= Grade 3 urinary toxicities is low. Nevertheless, especially among patients with a prior history of TURP who experience Grade 2 acute urinary symptoms during radiation treatment, a higher risk of stress incontinence is observed. PMID- 11020560 TI - Reduction of small and large bowel irradiation using an optimized intensity modulated pelvic radiotherapy technique in patients with prostate cancer. AB - PURPOSE: To investigate the role of intensity-modulated radiation therapy (IMRT) to irradiate the prostate gland and pelvic lymph nodes while sparing critical pelvic organs, and to optimize the number of beams required. METHODS AND MATERIALS: Target, small bowel, colon, rectum, and bladder were outlined on CT planning scans of 10 men with prostate cancer. Optimized conventional (RT) and 3 dimensional conformal radiotherapy (3D-CRT) plans were created and compared to inverse-planned IMRT dose distributions using dose-volume histograms. Optimization of beam number was undertaken for the IMRT plans. RESULTS: With RT the mean percentage volume of small bowel and colon receiving >45 Gy was 21.4 +/- 5.4%. For 3D-CRT it was 18.3 +/- 7.7% (p = 0.0043) and for 9-field IMRT it was 5.3 +/- 1.8% (p < 0.001 compared to 3D-CRT). For 7, 5, and 3 IMRT fields, it was 6.4 +/- 2.9%, 7.2 +/- 2.8%, and 8.4 +/- 3.8% (all p < 0.001 compared to 3D-CRT). The rectal volume irradiated >45 Gy was reduced from 50.5 +/- 16.3% (3D-CRT) to 5.8 +/- 2.1% by 9-field IMRT (p < 0. 001) and bladder from 52.2 +/- 12.8% to 7 +/ 2.8% (p < 0.001). Similar benefits were maintained for 7, 5, and 3 IMRT fields. CONCLUSIONS: The reduction in critical pelvic organ irradiation seen with IMRT may reduce side effects in patients, and allow modest dose escalation within acceptable complication rates. These reductions were maintained with 3-5 IMRT field plans which potentially allow less complex delivery techniques and shorter delivery times. PMID- 11020561 TI - A prospective comparison of three systems of patient immobilization for prostate radiotherapy. AB - PURPOSE: The study compared the setup reliability of 3 patient immobilization systems, a rubber leg cushion, the alpha cradle, and the thermoplastic Hipfix device, in 77 patients with cT1-T3, N0, M0 prostate cancer receiving conformal radiotherapy. METHODS AND MATERIALS: Port films were analyzed and compared to simulation films to estimate the setup errors in the three coordinate axes (anterior-posterior, cranial-caudal, medial-lateral). A total vector error was calculated from these shifts. RESULTS: The Hipfix was found significantly superior to the other two devices in reducing mean setup errors in all axes (p < 0.005). The average field-positioning error with the Hipfix ranged from 1.9 mm to 2.6 mm for all axes, whereas the deviation for the other two systems ranged from 2.7 to 3. 4 mm. Errors greater than 10 mm were virtually eliminated with the Hipfix system. There was a reduction in the mean total vector error in the alpha cradle and Hipfix patient cohorts over time, reflecting improved efficacy as a result of experience. CONCLUSION: There was a significant difference in the performance of each immobilization device. The Hipfix was consistently more reliable in reducing setup errors than the other devices. PMID- 11020562 TI - Rectal function following prostate brachytherapy. AB - PURPOSE: Quality of life following therapeutic intervention for carcinoma of the prostate gland has not been well documented. In particular, a paucity of data has been published regarding bowel function following prostate brachytherapy. This study evaluated late bowel function in 209 consecutive prostate brachytherapy patients via a one-time questionnaire administered 16-55 months postimplant. MATERIALS AND METHODS: Two hundred nineteen consecutive patients underwent permanent prostate brachytherapy from April 1995 through February 1998 using either (125)I or (103)Pd for clinical T1c-T3a carcinoma of the prostate gland. Of the 219 patients, 7 had expired. Of the remaining 212 patients (median follow-up, 28 months), each patient was mailed a self-administered questionnaire (10 questions) with a prestamped return envelope; 209 (98.6%) surveys were returned. Clinical parameters evaluated for bowel dysfunction included patient age, diabetes, hypertension, history of tobacco consumption, clinical T-stage, elapsed time since implant, and prostate ultrasound volume. Treatment parameters included utilization of neoadjuvant hormonal manipulation, utilization of moderate dose external beam radiation therapy prior to implantation, choice of isotope ((125)I vs. (103)Pd), rectal dose (average, median and maximum doses), total implanted seed strength, values of the minimum dose received by 90% of the prostate gland (D(90)), and the percent prostate volume receiving 100%, 150%, and 200% of the prescribed minimum peripheral dose (V(100), V(150) and V(200), respectively). Because detailed baseline bowel function was not available for these patients, a cross-sectional survey was performed in which 30 newly diagnosed prostate cancer patients of comparable demographics served as controls. RESULTS: The total rectal function scores for the brachytherapy and control patients were 4.3 and 1.6, respectively, out of a total 27 points (p < 0.001). Of the evaluated clinical parameters, only the preimplant number of bowel movements per day were correlated with the total survey score (p < 0.01). None of the treatment parameters were significantly correlated with the total survey score. Despite the fact that implantation with (103)Pd resulted in lower radiation doses to the rectum, the choice of isotope was not predictive of bowel function scores. A trend toward increased rectal scores was noted for older patients, and a nonsignificant improvement in rectal survey scores was noted with elapsed time from implantation. Only 19.2% (40/208) of the treatment group reported a worsening of bowel function following implantation. Patient perception of overall rectal quality of life, however, was inversely related to the utilization of external beam radiation therapy (p = 0.034). CONCLUSION: To date, no severe changes in late bowel function have been noted following prostate brachytherapy. Although the survey scores indicate bowel function is worse after an implant, the minor changes are not significant enough to bother most individuals. Less than 20% of patients reported that their bowel function was worse following prostate brachytherapy. PMID- 11020563 TI - High-dose-rate interstitial brachytherapy as a monotherapy for localized prostate cancer: treatment description and preliminary results of a phase I/II clinical trial. AB - PURPOSE: To improve results for localized prostate cancer, a prospective clinical trial of hyperfractionated Iridium-192 high-dose-rate (HDR) brachytherapy as a monotherapy was initiated. METHODS AND MATERIALS: Between May 1995 and September 1998, 22 implants were performed on 22 patients with localized prostate cancer (T1:T2:T3:T4 = 4:6:9:3) at Osaka University Hospital. Nineteen patients, who had T3-T4 tumors or pretreatment PSA >/= 20.0 ng/mL, received hormone therapy. No patient had external beam radiation. Transperineal needle implants using real time ultrasound guidance were performed, followed by dose optimization program. Patients were irradiated twice a day, with a time interval of more than 6 h. Total dose was 48 Gy/8 fractions/5 days or 54 Gy/9 fractions/5 days. Acute toxicity was scored using the Radiation Therapy Oncology Group (RTOG) radiation morbidity scoring criteria. Median follow-up time was 31 months. RESULTS: HDR brachytherapy as a monotherapy was well-tolerated. No significant intra- or peri operative complications occurred. No patient experienced acute toxicity of grade 3 or more. PSA levels normalized in 95% of patients within 20 months after irradiation. Four-year clinical and biochemical relapse-free rates were 95% and 55%, respectively. CONCLUSION: Acute toxicity with this method was acceptable. Further patient accrual and longer follow-up will allow comparison to other techniques. PMID- 11020564 TI - Feasibility and acute toxicities of radioimmunoguided prostate brachytherapy. AB - PURPOSE: We present a technique that fuses pelvic CT scans and ProstaScint images to localize areas of disease within the prostate gland to customize prostate implants. Additionally, the acute toxicity results from the first 43 patients treated with this technique are reviewed. METHODS AND MATERIALS: Between 2/97 and 8/98, 43 patients with clinical stage II prostate adenocarcinoma received ultrasound-guided transperineal implantation of I-125 or Pd-103 seeds. The median patient age was 70 years (range 49-79). Prior to treatment, the median Gleason score and prostate-specific antigen (PSA) were 6 (range 3-8) and 7.5 (range 1.8 16.6 ng/mL), respectively. The median follow-up was 10 months (range 2.9-20.4 months). RESULTS: The median PSA value at 10 months is 0.7 ng/mL. Significant acute complications within the first month following implantation included 13 Grade I urinary symptoms, 24 Grade II urinary symptoms, 6 Grade III symptoms, and no Grade IV complications. Beyond 4 months, complications included 12 Grade I urinary symptoms, 17 Grade II urinary symptoms, 1 Grade III, and 1 Grade IV complications. CONCLUSIONS: The image fusion of the pelvic CT scan and ProstaScint scans helped identify regions within the prostate at high risk of local failure, which were targeted with additional seeds during implantation. PMID- 11020565 TI - Predictors of severe esophagitis include use of concurrent chemotherapy, but not the length of irradiated esophagus: a multivariate analysis of patients with lung cancer treated with nonoperative therapy. AB - PURPOSE: To identify in a multivariate analysis treatment-related factors predisposing patients (pts) with lung cancer to acute esophagitis, expressed as a severity grade or Esophagitis Index (EI). METHODS AND MATERIALS: Acute esophagitis is prospectively scored as an RTOG Grade in our institution during and after thoracic radiotherapy. Charts, toxicity forms and digitally reconstructed radiographs (DRRs) of all pts with lung cancer who received thoracic radiotherapy (RT) between 11/95 and 1/99 were reviewed. Esophagitis grades for each time point were verified by review of weekly physician and nursing treatment notes, hospital discharge summaries and referring physician notes and then plotted on graph against time. The area under the curve was calculated for each patient's graph and was defined as an Esophagitis Index. The length of esophagus was measured on each anterior DRR while assuming that esophagus overlies the vertebral bodies on the anterior films and projects over the edge of the vertebral body on the oblique DRRs. This assumption was confirmed in 10 pts by digitizing esophagus on CT simulator-derived slices and visualizing its position on DRRs. To compare RT doses delivered with different fractionation schemes to standard fractionated doses, the equivalent RT doses were calculated using the linear-quadratic formula and alpha/beta ratio of 10. Univariate and multivariate analyses of several factors potentially influencing the maximum esophagitis grade, as well as EI, were performed. RESULTS: A total of 277 pts were identified. Pts were included in the analysis (n = 105) if they fulfilled the following criteria: chart, toxicity form and DRRs were all available; parallel opposed fields (no multiple fields) were used for both the initial and off cord/cone down fields; and an equivalent dose of 45.0 Gy or more was delivered. Seventy-eight pts had Stage III; 32, Stage IV, and the remainder, Stages I, II, or recurrent lung cancer (85 non-small cell and 18, small cell). Seventy-four pts were treated with definitive intent. Chemotherapy was given concurrently with RT in 58 pts (in 7 pts, with twice daily, or b.i.d., RT) and as induction treatment, in 11. Only 2 pts required a treatment break of more than 1 week. Median total and equivalent RT doses, fraction size, and anterior esophageal length were as follows: 59.9 Gy, 59.9 Gy, 2.0 Gy, and 14 cm (range, 4.2-21). The following maximum grades of esophagitis were recorded: 1, in 54 pts; 2, in 17 pts; 3, in 13 pts, and 4, in 1 pt. The mean EI for all pts; pts treated with standard RT alone; induction chemotherapy and standard RT; concurrent chemotherapy and standard (QD) RT; and b.i.d. RT with concurrent chemotherapy, was 41. 5 (range, 0-317); 13.6; 24.5; 52.4; and 132.1, respectively (p < 0. 001). Three pts developed an esophageal stricture within 3 months beginning RT. In multivariate analysis, the following factors were significantly associated with increasing EI: concurrent chemotherapy with QD RT and concurrent chemotherapy with b.i.d. RT (p < 0.001, considered jointly). Both factors were also associated with increasing maximum esophagitis grade (p = 0.011). Esophageal length was not associated with increasing EI or esophagitis grade in either univariate or multivariate analyses. CONCLUSION: Concurrent chemotherapy and twice daily radiotherapy, especially if combined together, were associated with the highest acute maximum esophagitis grade and esophagitis index in pts with lung cancer. The duration of acute esophagitis was also longest in the concurrent chemotherapy/twice daily radiotherapy group. Esophagitis Index appeared to be a more sensitive measure of acute esophagitis than the maximum esophagitis grade. The increasing length of esophagus in the radiation field did not predict for the severity of acute esophagitis. PMID- 11020566 TI - High-dose-rate endobronchial brachytherapy effectively palliates symptoms due to airway tumors: the 10-year M. D. Anderson cancer center experience. AB - PURPOSE: To evaluate the toxicity and efficacy of Iridium-192 high-dose-rate (HDR) endobronchial brachytherapy (EBBT) for the palliation of symptoms caused by relapsed or persistent endobronchial tumors. METHODS AND MATERIALS: We reviewed the treatment outcomes between 1988 and 1997 in 175 lung cancer patients who underwent HDR EBBT for recurrent or metastatic tumors at The University of Texas M. D. Anderson Cancer Center. One hundred sixty of these patients had previously received thoracic external-beam irradiation. This updated report includes 74 patients from a previous series. Most patients received 3,000-cGy EBBT delivered at a distance of 6 mm and divided into 2 fractions over 2 weeks. Subjective response was assessed by questionnaire at follow-up. Objective response was assessed by physical examination, bronchoscopy, and chest radiograph. RESULTS: The median actuarial survival for the entire group was 6 months from the time of the first EBBT treatment session. Of the 115 patients (66%) who showed symptomatic improvement, 32% were much improved and 34% were slightly improved. Patients showing improvement survived for significantly longer than those who showed no change or worsening symptoms (7 vs. 4 months, p = 0.0032). Repeat bronchoscopy demonstrated a 78% overall objective response rate that correlated significantly with subjective response and symptom relief. Complications occurred in 19 patients (11% crude rate) with an actuarial complication rate of 13% at 1 year from the time of the first EBBT treatment session. The actuarial hazard for fatal hemoptysis due to EBBT was 5%. CONCLUSION: HDR EBBT effectively palliates most patients' symptoms caused by endobronchial lesions. This relief correlates significantly with an overall survival benefit. Treatment complications appear to be few, even for patients who have received prior external-beam irradiation. PMID- 11020567 TI - Involved-field radiotherapy alone for early-stage non-small-cell lung cancer. AB - PURPOSE: To describe the outcome of involved-field radiotherapy in patients with early-stage non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: A written policy for the radical treatment of early-stage NSCLC with involved-field radiotherapy was adopted at our center in 1986. The sites of known disease were treated to a dose of 52.5 Gy in 20 daily fractions over 4 weeks without elective irradiation of radiologically uninvolved regional nodes. We have reviewed the outcome of this policy in 102 patients treated with radiotherapy alone between 1986 and 1995. RESULTS: The patients' median age was 71.5 years. The stage distribution was as follows: T1, 33.3%; T2, 56.9%; T3, 8.8%; and T4, 1.0%. Only 5 cases were N1; the remainder were N0. Most patients (76.5%) were not surgical candidates because of co-morbidity. Ninety-three percent had a CT of the thorax as part of their initial staging, while mediastinoscopy was performed in only 16.7%. Overall survival was 35% at 3 years and 16% at 5 years. Recurrence free survival was 23.9% at 3 years and 13. 9% at 5 years. Cause-specific survival was 43.5% at 3 years and 26. 8% at 5 years. Of those who recurred, 68.9% had a local component of failure at initial relapse, and 49.2% failed locally without evidence of regional or distant metastases. Isolated regional nodal relapse at initial failure occurred in only 6.6% of recurrences. There were no treatment interruptions due to acute toxicity and no treatment-related deaths. CONCLUSION: Involved-field radiotherapy alone cures a small but significant number of patients with early-stage NSCLC. This approach is recommended in patients who are unfit for surgery and who have severely compromised pulmonary function that would preclude the use of wide-field radiotherapy. The dose used in this study was well tolerated, but produced suboptimal local control rates. PMID- 11020568 TI - Three-dimensional intensity-modulated radiotherapy in the treatment of nasopharyngeal carcinoma: the University of California-San Francisco experience. AB - PURPOSE: To review our experience with three-dimensional intensity-modulated radiotherapy (IMRT) in the treatment of nasopharyngeal carcinoma. METHODS AND MATERIALS: We reviewed the records of 35 patients who underwent 3D IMRT for nasopharyngeal carcinoma at the University of California-San Francisco between April 1995 and March 1998. According to the 1997 American Joint Committee on Cancer staging classification, 4 (12%) patients had Stage I disease, 6 (17%) had Stage II, 11 (32%) had Stage III, and 14 (40%) had Stage IV disease. IMRT of the primary tumor was delivered using one of the following three techniques: (1) manually cut partial transmission blocks, (2) computer-controlled autosequencing static multileaf collimator (MLC), and (3) Peacock system using a dynamic multivane intensity-modulating collimator (MIMiC). A forward 3D treatment planning system was used for the first two methods, and an inverse treatment planning system was used for the third method. The neck was irradiated with a conventional technique using lateral opposed fields to the upper neck and an anterior field to the lower neck and supraclavicular fossae. The prescribed dose was 65-70 Gy to the gross tumor volume (GTV) and positive neck nodes, 60 Gy to the clinical target volume (CTV), and 50-60 Gy to the clinically negative neck. Eleven (32%) patients had fractionated high-dose-rate intracavitary brachytherapy boost to the primary tumor 1-2 weeks following external beam radiotherapy. Thirty two (91%) patients also received cisplatin during, and cisplatin and 5 fluorouracil after, radiotherapy. Acute and late normal tissue effects were graded according to the Radiation Therapy Oncology Group (RTOG) radiation morbidity scoring criteria. Local-regional progression-free, distant metastasis free survival and overall survival were estimated using the Kaplan-Meier method. RESULTS: With a median follow-up of 21.8 months (range, 5-49 months), the local regional progression-free rate was 100%. The 4-year overall survival was 94%, and the distant metastasis-free rate was 57%. The worst acute toxicity was Grade 2 in 16 (46%) patients, Grade 3 in 18 (51%) patients and Grade 4 in 1 (3%) patient. The worst late toxicity was Grade 1 in 15 (43%), Grade 2 in 13 (37%), and Grade 3 in 5 (14%) patients. Only 1 patient had a transient Grade 4 soft-tissue necrosis. At 24 months after treatment, 50% of the evaluated patients had Grade 0, 50% had Grade 1, and none had Grade 2 xerostomia. Analysis of the dose-volume histograms (DVHs) showed that the average maximum, mean, and minimum dose delivered were 79.5 Gy, 75.8 Gy, and 56.5 Gy to the GTV, and 78.9 Gy, 71.2 Gy, and 45.4 Gy to the CTV, respectively. An average of only 3% of the GTV and 2% of the CTV received less than 95% of the prescribed dose. The average dose to 5% of the brain stem, optic chiasm, and right and left optic nerves was 48.3 Gy, 23.9 Gy, 15.0 Gy, and 14.9 Gy, respectively. The average dose to 1 cc of the cervical spinal cord was 41.7 Gy. The doses delivered were within the tolerance of these critical normal structures. The average dose to 50% of the right and left parotids, pituitary, right and left T-M joints, and ears was 43. 2 Gy, 41.0 Gy, 46.3 Gy, 60.5 Gy, 58.3 Gy, 52.0 Gy, and 52.2 Gy, respectively. CONCLUSION: 3D intensity-modulated radiotherapy provided improved target volume coverage and increased dose to the gross tumor with significant sparing of the salivary glands and other critical normal structures. Local-regional control rate with combined IMRT and chemotherapy was excellent, although distant metastasis remained unabated. PMID- 11020569 TI - Radiation therapy in T1-T2 glottic carcinoma: influence of various treatment parameters on local control/complications. AB - PURPOSE: To evaluate the influence of various treatment parameters on local control as well as complications in T1 and T2 glottic carcinomas. METHODS AND MATERIALS: Between 1975 and 1989, 676 patients with early glottic carcinoma (460 T1 and 216 T2) received curative radiation with three different treatment regimens, as follows: Regimen 1-50 Gy/15 Fr/3 weeks (3.33 Gy/daily) for 192 patients; Regimen 2-60-62.5 Gy/24-25 Fr/5 weeks (2.5 Gy/daily) for 352 patients; and Regimen 3-55-60 Gy/25-30 Fr/5-6 weeks (2-2.25 Gy/daily) for 132 patients. RESULTS: The local control at 10 years was 82% and 57% for T1 and T2 lesions respectively (p = 0.0). For the T1N0M0 group, field size had significant impact on local control with both univariate (p = 0.05) and multivariate (p = 0.03) analysis. For T2N0M0, group field size (p = 0.03) as well as registration year (p = 0.016) were significant in univariate analysis whereas only field size remained significant on multivariate analysis. Persistent radiation edema was noted in 146 (22%) patients and was significantly worse with larger field size (p = 0.000) but not related to different treatment regimens. CONCLUSION: The shorter fractionation schedule had comparable local control, without increased complications in comparison to the protracted schedule and is best suited for a busy department. PMID- 11020570 TI - An analysis of facial nerve function in irradiated and unirradiated facial nerve grafts. AB - PURPOSE: The effect of high-dose radiation therapy on facial nerve grafts is controversial. Some authors believe radiotherapy is so detrimental to the outcome of facial nerve graft function that dynamic or static slings should be performed instead of facial nerve grafts in all patients who are to receive postoperative radiation therapy. Unfortunately, the facial function achieved with dynamic and static slings is almost always inferior to that after facial nerve grafts. In this retrospective study, we compared facial nerve function in irradiated and unirradiated nerve grafts. METHODS AND MATERIALS The medical records of 818 patients with neoplasms involving the parotid gland who received treatment between 1974 and 1997 were reviewed, of whom 66 underwent facial nerve grafting. Fourteen patients who died or had a recurrence less than a year after their facial nerve graft were excluded. The median follow-up for the remaining 52 patients was 10.6 years. Cable nerve grafts were performed in 50 patients and direct anastomoses of the facial nerve in two. Facial nerve function was scored by means of the House-Brackmann (H-B) facial grading system. Twenty-eight of the 52 patients received postoperative radiotherapy. The median time from nerve grafting to start of radiotherapy was 5.1 weeks. The median and mean doses of radiation were 6000 and 6033 cGy, respectively, for the irradiated grafts. One patient received preoperative radiotherapy to a total dose of 5000 cGy in 25 fractions and underwent surgery 1 month after the completion of radiotherapy. This patient was placed, by convention, in the irradiated facial nerve graft cohort. RESULTS: Potential prognostic factors for facial nerve function such as age, gender, extent of surgery at the time of nerve grafting, preoperative facial nerve palsy, duration of preoperative palsy if present, or number of previous operations in the parotid bed were relatively well balanced between irradiated and unirradiated patients. However, the irradiated graft group had a greater proportion of patients with pathologic evidence of nerve invasion (p = 0.007) and unfavorable type of nerve graft (p = 0.04). Although the irradiated graft cohort had more potentially negative prognostic factors, there was no difference in functional outcome (H-B Grade III or IV) between irradiated and unirradiated graft patients. H-B Grades III, IV, V, and VI were the best postoperative facial nerve functions achieved in 35%, 39%, 13%, and 13% of patients, respectively. The patient with preoperative radiotherapy never recovered any facial nerve function (H-B Grade VI). Median time to best facial nerve function after surgery was longer in the irradiated patients (13.1 vs. 10.8 months), but this was not statistically significant (p = 0.10). Presence of preoperative facial nerve palsy (p = 0.005), duration of preoperative palsy (p = 0.003), and age greater than 60 years at the time of grafting (p = 0. 04) were all negative prognostic factors for achieving a functional facial nerve on univariate analysis. Analysis of age as a continuous variable (p = 0.12) and pathologic evidence of nerve invasion (p = 0. 1) revealed a trend toward negative prognostic factors. Gender, number of previous operations in the parotid bed, extent of surgery at the time of nerve grafting, and type of grafting procedure were not significant prognostic factors. Whether radiotherapy was delivered less than 6 weeks after nerve grafting or more than 6 weeks had no impact on achievement of a functional facial nerve. CONCLUSION: Negative prognostic factors for achieving a functional facial nerve in our series include the presence of preoperative facial nerve palsy, duration of preoperative palsy, and age greater than 60 years. Radiotherapy was not a negative prognostic factor. Comparing irradiated and unirradiated grafts revealed no difference in best facial nerve function achieved, despite the presence of a greater proportion of negative prognostic factors in PMID- 11020571 TI - Timescale of evolution of late radiation injury after postoperative radiotherapy of breast cancer patients. AB - PURPOSE: To evaluate the incidence and prevalence of various signs of late morbidity, their time of appearance and pattern of progression during an observation period up to 34 years in breast cancer patients treated with postoperative radiation therapy after radical mastectomy. METHODS AND MATERIALS: A group of 71 breast cancer patients received in 1963-1965 aggressive postoperative telecobalt therapy to the parasternal, axillary, and supraclavicular lymph node regions after total mastectomy and axillary clearance. None of the patients received chemotherapy either prior to, or after the irradiation as part of their primary treatment. The prescribed dose to the three lymph node regions was 44 Gy in 11 fractions. Only two of the three fields were treated per day. This total dose was given in 16-17 fractions over 3-4 weeks. Because of the overlap of the supraclavicular and axillary fields, the dose received by the brachial plexus was not the dose that was prescribed. A retrospective dose calculation showed that the total dose to the brachial plexus was 57 Gy, delivered as a complex combination of 1.8 Gy, 3.4 Gy, and 5.2 Gy fractions. This cohort of patients has now been followed to 34 years and the late side effects of the treatment evaluated and scored. RESULTS: This series is unique in the literature. There is no comparable report of a detailed long-term follow-up in a homogeneously treated group of patients with such a high survival, especially among the younger women, where it is almost 50% at 30 years. This is the reason that they were able to develop some of the very slowly evolving injuries. There was progression of many of the late effects in the period between 5 and 34 years. The more serious morbidities have increased progressively over the whole 34-year follow-up period. Ninety-two percent of the long-term survivors have paralysis of their arm. Other neurological findings included unilateral vocal cord paralysis among 5% of the patients, who developed the disease after a median time of 19 years. All of them were left-sided, indicating a mediastinal involvement of the recurrent nerve. Local recurrence or the appearance of a new primary tumor infiltrating or causing pressure on the recurrent nerve were vigorously investigated and excluded as possible causes of these symptoms. CONCLUSION: The greatest risk for all cancer patients is the inadequate treatment of their disease, because this is inevitably lethal. The aggressiveness of the therapy and the acceptable risk of complications must therefore be balanced against the risk of recurrence. The neuropathy seems to be closely linked to the development of fibrosis around the nerve trunks. The use of large daily fractions, combined with hot spots from overlapping fields contributed to the severity of the complications. PMID- 11020572 TI - Bronchiolitis obliterans organizing pneumonia syndrome in breast-conserving therapy for early breast cancer: radiation-induced lung toxicity. AB - PURPOSE: Breast-conserving surgery and postoperative radiotherapy have played important roles in the treatment of early breast cancer. Bronchiolitis obliterans organizing pneumonia (BOOP) syndrome has recently been reported to be one of the complications of adjuvant radiotherapy. The purpose of this study was to determine the incidence of and risk factors for BOOP syndrome in breast cancer patients. METHODS AND MATERIALS: Between January 1996 and December 1998, 157 patients with breast cancer underwent radiotherapy after breast-conserving surgery. The criteria used for the diagnosis of BOOP syndrome were as follows: 1) radiation therapy to the breast within 12 months, 2) general and/or respiratory symptoms lasting for at least 2 weeks, 3) radiographic lung infiltrates outside the radiation port, and 4) no evidence of a specific cause. RESULTS: BOOP syndrome developed in 4 (2.5%) patients, who had fever and nonproductive cough, with patchy infiltrative shadows on chest roentgenograms which emerged between 5 and 6 months after radiotherapy. The symptoms and pulmonary infiltrates were rapidly improved by treatment with prednisone (40 mg/day), which was tapered over 2- to 5-month periods. However, BOOP syndrome relapsed in all cases during the tapering period or after withdrawal of prednisone. The eosinophil and neutrophil counts were increased and the ratios of CD4+ to CD8+ lymphocytes were elevated in bronchoalveolar lavage fluid in all four cases. There were no differences in proportions of patients by age, irradiated breast site, use of tamoxifen and/or chemotherapy, or radiation dose between those with and without BOOP syndrome. CONCLUSIONS: BOOP syndrome is considered an intractable form of lung toxicity after radiotherapy to the breast. An immunologic reaction mediated by eosinophils, neutrophils, and lymphocytes may be responsible for the development of this syndrome. Methods of prevention of BOOP syndrome should be established. PMID- 11020573 TI - External beam irradiation plus (192)Ir implant after breast-preserving surgery in women with early breast cancer. AB - PURPOSE: To provide more information for the clinician and to analyze the impact of the boost with brachytherapy on the local disease-free survival (LDFS), disease-free survival (DFS), specific overall survival (OS), and cosmesis, a retrospective study of external radiation therapy and (192)Ir implantation in early breast cancer at Institut Catala d'Oncologia has been undertaken. PATIENTS AND METHODS: From 1986 to 1996, 530 patients were selected for this study with a median follow-up period of 39.5 months (range, 10-115 months). External radiation therapy (combined with brachytherapy) was administered postoperatively to the breast in all patients. Mean given dose was 48.7 Gy (range, 42-52 Gy) with external radiation therapy to the breast, and 16.8 Gy (range, 10-27 Gy) was the mean dose with brachytherapy. Variables have been tested for cosmesis. Univariate and multivariate analysis have also been carried out. RESULTS: Mean age of the patients was 54 years (range, 28-81 years). Stages were distributed as follows: 350 patients (66%) in Stage I, 173 in Stage II (32.8%), and 7 in Stage III (1.1%). Pathologic distribution was 445 patients with infiltrating ductal carcinoma (84%), 20 patients with infiltrating lobular carcinoma (4%), and 65 patients (12%) of a miscellaneous group. OS for the entire group was 89.4% and 85.9% at 5 and 7 years respectively. Probability of DFS was 81.7% and 70.1% at 5 and 7 years. The LDFS was 94.9% and 91.7% at 5 and 7 years. The MFS probability was 85.5% and 76.9% at 5 and 7 years, respectively. Univariate analysis demonstrated that age (older than 52 years), premenopausal status, moderate and high histologic grades (Grades II-III), and presence of intraductal comedocarcinoma were prognostic factors for local relapse. Multivariate analysis for local disease-free survival demonstrated that only perineural or muscular infiltration remain as prognostic factors. Tumor dose bed of 70 Gy or higher had a negative impact in breast subcutaneous fibrosis, whereas dose rate lower than 65 cGy/h was better for skin color at 2 years. CONCLUSION: We conclude that patients with early-stage breast cancer undergoing external radiotherapy and LDR brachytherapy can be effectively managed. Overall survival, long-term local control, and cosmetic control are excellent. PMID- 11020574 TI - Treatment of high-risk uterine cancer with whole abdominopelvic radiation therapy. AB - PURPOSE: To evaluate the treatment outcomes in patients with optimally debulked Stage III and IV endometrial adenocarcinoma (ACA) or Stages I-IV uterine papillary serous (UPSC) or clear cell (CCC) carcinoma of the uterus, treated postoperatively with whole abdominopelvic irradiation (WAPI). METHODS AND MATERIALS: Between 1979 and 1998, 48 patients received postoperative WAPI at our institution. Twenty-two patients had FIGO Stage III or Stage IV ACA and 26 patients had FIGO Stages I-IV UPSC or CCC. The median dose was 30 Gy to the upper abdomen and 49.8 Gy to the pelvis. Mean follow-up was 37 months (2.4-135 months). RESULTS: The 3-year estimated disease-free survival (DFS) and overall survival (OS) rates for the entire group were 60% and 77%, respectively. Patients with ACA had 3-year DFS and OS of 79% and 89%, respectively, compared with 47% and 68% in the UPSC/CCC group. Early-stage patients (I and II) with UPSC/CCC had 3-year DFS and OS of 87% compared with 32% and 61% in those with advanced (Stage III and IV) disease. The 3-year actuarial major complication rate was 7%, with no treatment related deaths. All 4 failures in the ACA group were extra-abdominal and 6 of the 11 in the UPSC/CCC group had an extra-abdominal component. Age and UPSC/CCC histology were significant prognostic factors for DFS and OS. In addition, stage and number of extrauterine sites of disease were significant predictors for DFS in UPSC/CCC. CONCLUSION: WAPI is a safe, effective treatment for patients with optimally debulked advanced-stage uterine ACA or early-stage UPSC/CCC. Survival was significantly worse in advanced-stage UPSC/CCC patients. We recommend future trials of WAPI with concurrent, or subsequent systemic therapy in patients with advanced-stage UPSC or CCC. PMID- 11020575 TI - The American Brachytherapy Society recommendations for high-dose-rate brachytherapy for carcinoma of the endometrium. AB - PURPOSE: To develop recommendations for use of high-dose-rate (HDR) brachytherapy in patients with endometrial cancer. METHODS: A panel of members of the American Brachytherapy Society (ABS) performed a literature review, supplemented their clinical experience, and formulated recommendations for endometrial HDR brachytherapy. RESULTS: The ABS endorses the National Comprehensive Cancer Network (NCCN) guidelines for indications for radiation therapy for patients with endometrial cancer and the guidelines on HDR quality assurance of the American Association on Physicists in Medicine (AAPM). The ABS made specific recommendations for HDR applicator selection, insertion techniques, target volume definition, dose fractionation, and specifications for postoperative adjuvant vaginal cuff therapy, for vaginal recurrences, and for medically inoperable primary endometrial cancer patients. The ABS recommends that applicator selection should be based on patient and target volume geometry. The dose prescription point should be clearly specified. The treatment plan should be optimized to conform to the target volume whenever possible while recognizing the limitations of computer optimization. Suggested doses were tabulated for treatment with HDR alone, and in combination with external beam radiation therapy (EBRT), when applicable. For intravaginal brachytherapy, the largest diameter applicator should be selected to ensure close mucosal apposition. Doses should be reported both at the vaginal surface and at 0.5-cm depth irrespective of the dose prescription point. For vaginal recurrences, intracavitary brachytherapy should be restricted to patients with nonbulky (< 0.5-cm thick) disease. Patients with bulky (> 0.5-cm thick) recurrences should be treated with interstitial techniques. For medically inoperable patients, an appropriate applicator that will allow adequate irradiation of the entire uterus should be selected. CONCLUSION: Recommendations are made for HDR brachytherapy for endometrial cancer. Practitioners and cooperative groups are encouraged to use these recommendations to formulate their treatment and dose reporting policies. This will lead to meaningful comparisons of reports from different institutions and lead to advances and appropriate use of HDR. PMID- 11020576 TI - A phase I/II evaluation of tirapazamine administered intravenously concurrent with cisplatin and radiotherapy in women with locally advanced cervical cancer. AB - PURPOSE: This is a Phase I/II dose escalation study to determine the tolerable dose of tirapazamine (TPZ), and the toxicity of a regimen using TPZ with cisplatin, and radiotherapy in women with locally advanced cervical cancer. METHODS AND MATERIALS: Eligible women for this study were those with a diagnosis of locally advanced cervix cancer, who were less than 75 years of age, having provided informed consent, and who had undergone the necessary prestudy investigations. External-beam radiotherapy (RT) was given to a minimum dose of 4500 cGy in 25 fractions (Day 1-35), and brachytherapy then delivered to bring the total dose at point A to 8500 cGy. The first dose level of the study used TPZ 190 mg/m(2) and cisplatin 75 mg/m(2) on Days 1, 15, and 29 of RT. TPZ 160 mg/m(2) alone was used on Days 8, 10, 12 and 22, 24, 26 of RT. A conventional dose escalation step method was then used to determine the maximum tolerated dose (MTD) of TPZ. RESULTS: Four patients were treated at Level 1, 6 at Level 2, and 5 at Level 3. Only 1 patient experienced a dose-limiting toxicity (DLT) at Level 2, but 2 of the 5 patients at Level 3 incurred DLTs. Level 2 was declared the MDT (TPZ 290 mg/m(2) on Days 1, 15, 29 and 220 mg/m(2) on Days 8, 10, 12 and 22, 24, 26). At 6 months, 13 of 15 patients had complete pelvic control of disease. CONCLUSION: Level 2 of this regime was identified as the MDT. The use of TPZ with concurrent cisplatin and pelvic radiotherapy has acceptable toxicity and should be considered for further Phase 2 testing in view of the promising responses noted. PMID- 11020577 TI - Active breathing control (ABC) for Hodgkin's disease: reduction in normal tissue irradiation with deep inspiration and implications for treatment. AB - PURPOSE: Active breathing control (ABC) temporarily immobilizes breathing. This may allow a reduction in treatment margins. This planning study assesses normal tissue irradiation and reproducibility using ABC for Hodgkin's disease. METHODS AND MATERIALS: Five patients underwent CT scans using ABC obtained at the end of normal inspiration (NI), normal expiration (NE), and deep inspiration (DI). DI scans were repeated within the same session and 1-2 weeks later. To simulate mantle radiotherapy, a CTV1 was contoured encompassing the supraclavicular region, mediastinum, hila, and part of the heart. CTV2 was the same as CTV1 but included the whole heart. CTV3 encompassed the spleen and para-aortic lymph nodes. The planning target volume (PTV) was defined as CTV + 9 mm. PTVs were determined at NI, NE, and DI. A composite PTV (comp-PTV) based on the range of NI and NE PTVs was determined to represent the margin necessary for free breathing. Lung dose-mass histograms (DMH) for PTV1 and PTV2 and cardiac dose-volume histograms (DVH) for PTV3 were compared at the three different respiratory phases. RESULTS: ABC was well-tolerated by all patients. DI breath-holds ranged from 34 to 45 s. DMHs determined for PTV1 revealed a median reduction in lung mass irradiated at DI of 12% (range, 9-24%; n = 5) compared with simulated free breathing. PTV2 comparisons also showed a median reduction of 12% lung mass irradiated (range, 8-28%; n = 5). PTV3 analyses revealed the mean volume of heart irradiated decreased from 26% to 5% with deep inspiration (n = 5). Lung volume comparisons between intrasession and intersession DI studies revealed mean variations of 4%. CONCLUSION: ABC is well tolerated and reproducible. Radiotherapy delivered at deep inspiration with ABC may decrease normal tissue irradiation in Hodgkin's disease patients. PMID- 11020578 TI - Cataract-free interval and severity of cataract after total body irradiation and bone marrow transplantation: influence of treatment parameters. AB - PURPOSE: To determine prospectively the cataract-free interval (latency time) after total body irradiation (TBI) and bone marrow transplantation (BMT) and to assess accurately the final severity of the cataract. METHODS AND MATERIALS: Ninety-three of the patients who received TBI as a part of their conditioning regimen for BMT between 1982 and 1995 were followed with respect to cataract formation. Included were only patients who had a follow-up period of at least 23 months. TBI was applied in one fraction of 8 Gy or two fractions of 5 or 6 Gy. Cataract-free period was assessed and in 56 patients, who could be followed until stabilization of the cataract had occurred, final severity of the cataract was determined using a classification system. With respect to final severity, two groups were analyzed: subclinical low-grade cataract and high-grade cataract. Cataract-free period and final severity were determined with respect to type of transplantation, TBI dose, and posttransplant variables such as graft versus host disease (GVHD) and steroid treatment. RESULTS: Cataract incidence of the analyzed patients was 89%. Median time to develop a cataract was 58 months for autologous transplanted patients. For allogeneic transplanted patients treated or not treated with steroids, median times were 33 and 46 months, respectively. Final severity was not significantly different for autologous or allogeneic patients. In allogeneic patients, however, final severity was significantly different for patients who had or had not been treated with steroids for GVHD: 93% versus 35% high-grade cataract, respectively. Final severity was also different for patients receiving 1 x 8 or 2 x 5 Gy TBI, from patients receiving 2 x 6 Gy as conditioning therapy: 33% versus 79% high-grade cataract, respectively. The group of patients receiving 2 x 6 Gy comprised, however, more patients with steroid treatment for GVHD. So the high percentage of high-grade cataract in the 2 x 6 Gy group might also have been caused to a significant extent by steroid treatment. The percentage of patients with high-grade cataract was lower in allogeneic transplanted patients without steroid treatment for GVHD than in autologous transplanted patients: 35% versus 48%. An explanation for this could be pretransplant therapy containing high-dose steroids. CONCLUSIONS: After high-dose rate TBI in one or two fractions, steroids for GVHD influence latency time of a cataract and are of great importance for the severity the cataract finally attains. Although a cataract will develop in all patients, a clinically important high-grade cataract is relatively infrequent in patients not treated with steroids. Pretransplant therapy might also influence final severity of cataract. PMID- 11020579 TI - The rationale and technique of staged-volume arteriovenous malformation radiosurgery. AB - PURPOSE: Stereotactic radiosurgery is an effective management strategy for properly selected arteriovenous malformation (AVM) patients. However, the risk of postradiosurgical radiation-related injury generally limits this procedure to patients with AVMs of an average diameter of 3 cm or less. Radiosurgery of large AVMs in a planned staged fashion was undertaken to limit the radiation exposure to the surrounding normal brain. METHODS AND MATERIALS: Between April 1997 and December 1999, 10 patients with a median AVM volume of 17.4 cm(3) (range, 7.4 53.3 cm(3)) underwent staged-volume radiosurgery (23 procedures). At the first radiosurgical procedure, the total volume of the AVM is estimated and a dose plan calculated that covers 10 cm(3)-15 cm(3), or one-half the nidus volume if the AVM is critically located (brainstem, thalamus, or basal ganglia). At 6-month intervals thereafter, radiosurgery was repeated to different portions of the AVM with the previous dose plan(s) being re-created utilizing intracranial landmarks to minimize radiation overlap. Radiosurgical procedures were continued until the entire malformation has been irradiated. RESULTS: The radiation dosimetry of staged-volume AVM radiosurgery was compared to hypothetical single-session procedures for the 10 patients. Staged-volume radiosurgery decreased the 12-Gy volume by an average of 11.1% (range, 4.9-21%) (p < 0.001). The non-AVM 12-Gy volume was reduced by an average of 27.2% (range, 12.5-51.3%) (p < 0.001). DISCUSSION: Staged-volume radiosurgery of large AVMs results in less radiation exposure to the adjacent brain. Further follow-up is needed to determine whether this technique provides a high rate of AVM obliteration while maintaining an acceptable rate of radiation-related complications. PMID- 11020580 TI - Allelic loss of chromosome 1p and radiotherapy plus chemotherapy in patients with oligodendrogliomas. AB - INTRODUCTION: Allelic loss of the short arm of chromosome 1 predicts radiographic response to chemotherapy and long overall survival times in patients with anaplastic oligodendrogliomas. Using a database of patients with oligodendrogliomas in whom chromosome 1p status was known, we explored whether allelic loss of 1p also predicted longer duration of tumor control when radiotherapy was part of the initial treatment of these patients. MATERIALS AND METHODS: We measured progression-free survival following radiotherapy in a cohort of patients with World Health Organization (WHO) Grade II and WHO Grade III oligodendrogliomas. The effects on progression-free survival of patient age, Karnofsky performance score (KPS), tumor grade when irradiated and chromosome 1p status were examined by univariate and multivariate statistical analyses. For the subset of patients with newly diagnosed anaplastic oligodendrogliomas, relationships between use of chemotherapy, chromosome 1p status and progression free survival were also examined. RESULTS: Fifty-five patients (29 male, 26 female; ages 18-75 years; median, 44 years; KPS 50-90, median 80) were irradiated for either a WHO Grade II (n = 19) or Grade III (n = 36) oligodendroglioma. Twenty-eight patients had chemotherapy immediately prior to radiotherapy, and 27 had chemotherapy at progression following radiotherapy. The median radiation dose was 54 Gy in 30 fractions. Loss of heterozygosity (LOH) at chromosome 1p was evident in 36 tumors and absent in 19. Overall median progression-free survival after radiotherapy was 40.4 months. Median progression-free survival was 55.0 months for patients whose tumors harbored 1p loss vs. 6.2 months for those patients whose tumors retained both copies of chromosome 1p (p < 0.001). On both univariate and multivariate analyses, chromosome lp loss was the principal independent predictor of longer progression-free survival for patients with Grade II and III oligodendrogliomas. For Grade III oligodendrogliomas, chemotherapy as an adjunct to radiotherapy prolonged tumor control for those patients whose tumors harbored allelic loss of chromosome 1p (p = 0.004). CONCLUSION: These data suggest allelic loss of chromosome 1p in patients with oligodendroglial neoplasms predicts longer progression-free survival among patients receiving radiotherapy +/- chemotherapy as part of their initial treatment. Chromosome 1p loss may be an important stratification variable in future therapeutic trials of oligodendroglioma. PMID- 11020581 TI - Extraneural metastatic glioblastoma after interstitial brachytherapy. AB - PURPOSE: This is a report of 3 cases of extraneural metastasis of glioblastoma after interstitial radiation and assessment of pertinent literature addressing concern over an increased risk of these events with this therapy. METHODS AND MATERIALS: In a series of 82 patients treated with (125)I brachytherapy for primary malignant brain tumors over a 7-year interval, 3 cases of extraneural glioblastoma were identified. The multicatheter technique for delivery of (125)I sources was utilized in all. Extraneural metastases were documented by imaging studies or biopsy. Over the same period, 310 patients with primary malignant brain tumors were treated without brachytherapy. RESULTS: Biopsy-proven scalp and skull metastases occurred in 2 patients, at 3 and 8 months following brachytherapy. Each developed radiographic evidence of systemic metastases at 7 and 14 months postbrachytherapy, respectively. The third patient developed biopsy proven cervical node involvement 4 months after brachytherapy. No patients with malignant gliomas undergoing craniotomy or stereotactic biopsy, but not brachytherapy, during the same time period developed extraneural metastases. Incidence in previously reported series commenting on this otherwise rare process range from 0% to 4.3%. The incidence of extraneural metastases in this series is 3.7% (3/82) and is comparable to those reports. CONCLUSIONS: Percutaneous catheter-delivered brachytherapy may be associated with an increased incidence of extraneural metastatic glioma. PMID- 11020582 TI - A retrospective analysis of 52 cases of spinal cord glioma managed with radiation therapy. AB - PURPOSE: To describe the outcome of primary spinal cord glioma treated with radiation therapy after surgery and to identify variables predictive of outcome. METHODS AND MATERIALS: A chart review of 52 patients with a diagnosis of spinal cord non-ependymoma glioma at the Princess Margaret Hospital was conducted. Thirty-two patients (62%) were male and 20 (38%) were female. Median age was 32 years (2-76 years). Median follow-up was 3.7 years (2 months to 27 years). Initial surgical management consisted of biopsy alone in 27 (52%) cases, subtotal resection in 20 (38%) cases, and gross total resection in 5 (10%) cases. All patients received postoperative radiation therapy; median total dose was 50 Gy, given in 25 daily fractions (20-60 Gy). Actuarial survival rates were calculated and the influence of patient-, tumor-, and treatment-related variables on outcome was determined. RESULTS: Five-year overall, cause-specific, and progression-free survivals were 54%, 62%, and 58%, respectively. Ten-year survivals were 45%, 50%, and 43%, respectively. A total of 29 (56%) patients died during the period of review. For 23 (79%) of these patients, death was cancer specific. Progression of tumor was documented in 28 of 52 (54%) patients. The following factors predicted for improved outcome on univariate analysis: age < 18 years, low-grade histology, and length of symptoms prior to diagnosis > 6 months. CONCLUSION: The outcome of patients in this series is consistent with that of other similar published reports. Specific recommendations are made for the management of this tumor. PMID- 11020583 TI - Preoperative chemotherapy and pelvic radiation for tethered or fixed rectal cancer: a phase II dose escalation study. AB - PURPOSE: To study the impact of preoperative radiation dose escalation and postoperative adjuvant chemotherapy on the outcome of tethered and fixed rectal carcinoma. METHODS AND MATERIALS: We have treated 156 patients with 3 consecutive preoperative chemoradiation protocols with escalating treatment intensity. Schedule 1 consisted of 40 Gy radiation with concurrent 5-fluorouracil (5-FU) infusion and mitomycin C. Schedule 2 used a sandwich design with preoperative (40 Gy) and postoperative (18 Gy) radiation with concomitant 5-FU infusion, leucovorin, and mitomycin C. In schedule 3, the preoperative radiation dose was increased to 50 Gy and adjuvant 5-FU/leucovorin chemotherapy was added following surgery. There were 54, 27, and 75 patients treated in schedules 1, 2, and 3, respectively. RESULTS: The resectability was 91% for schedule 1 and 100% for both schedules 2 and 3. A dose-response relationship was observed between the radiation dose and the tumor downstaging and local control. The pathological complete response (T0N0M0) rates for schedules 1, 2, and 3 were 4%, 15%, and 25%, respectively. The respective rates of tumor downstaging were 41%, 33%, and 68%, respectively. The 5-year local relapse-free rates were 67% for schedule 1 (40 Gy), 96% for schedule 2 (58 Gy), and 92% for schedule 3 (50 Gy) (p = 0.0011). The addition of postoperative chemotherapy appeared to improve both the survival and the relapse-free survival. The 5-year survival was increased from 52% to 84% (p = 0.0004) and the 5-year progression-free survival was improved from 48% to 74% (p = 0.0008). CONCLUSION: Preoperative 5-FU infusion, leucovorin, mitomycin C, and 50-Gy pelvic radiation, followed by postoperative bolus 5-FU/leucovorin chemotherapy, appeared to be an effective treatment for tethered/fixed rectal cancers. However, its therapeutic efficacy could only be validated in randomized studies. PMID- 11020584 TI - Clinical outcomes of orbital irradiation combined with or without systemic high dose or pulsed corticosteroids for Graves' ophthalmopathy. AB - PURPOSE: To evaluate the outcomes of orbital irradiation with or without high dose or pulsed corticosteroids in patients with Graves' ophthalmopathy (GO). METHODS AND MATERIALS: One hundred and twenty-one patients with moderate to severe GO who received orbital irradiation from 1987 to 1997 were retrospectively analyzed. A total dose of 20 Gy in 10 fractions was delivered to the bilateral retrobulbar volume. Eighty-six patients were treated in combination with high dose or pulsed corticosteroids and irradiation. Univariate and multivariate analyses were performed to assess the prognostic variables. RESULTS: The median follow-up period was 26 months. The overall clinical response was evaluated as excellent in 17 patients (14%), good in 65 (54%), fair in 31 (25%), no response in 7 (6%), and worse in 1 (1%). The best responses were noted for soft-tissue signs, extraocular muscle involvement, and sight loss, while a limited response was noted for proptosis. Multivariate analysis revealed that the use of high-dose corticosteroid or pulsed corticosteroids, female gender, and a shorter duration of ophthalmopathy before radiotherapy were significantly correlated with favorable outcomes. No long-term complications related to radiotherapy were observed. CONCLUSION: Orbital irradiation combined with high-dose or pulsed corticosteroids is an effective treatment for moderate to severe GO, especially in cases with major manifestations of soft-tissue signs, extraocular muscle impairment, or sight loss. PMID- 11020585 TI - Postoperative irradiation of primary or recurrent pterygium: results and sequelae. AB - PURPOSE: To evaluate the efficacy of postoperative beta irradiation and to analyze treatment sequelae in patients with primary and recurrent pterygium. METHODS AND MATERIALS: From June 1986 to June 1998, 94 patients corresponding to 100 eyes received postoperative beta irradiation. Two groups of patients were treated: 37 eyes with primary pterygium (Group I) and 63 eyes with recurrent pterygium (Group II). Terson technique surgery was used in the majority of patients. Time between surgery and beta irradiation ranged from 2 to 48 h. Radiation doses and fractionation consisted of 30 Gy/3 fractions/5 days in 17 cases, 60 Gy/6 fractions/6 weeks in 80 cases, and 20 Gy/1 fraction in 3 patients. RESULTS: Fourteen of the 100 cases (14%) treated with surgery and adjuvant irradiation recurred. The overall crude local recurrence rates were 5.4% for Group I and 19% for Group II patients. The 5-year probability of local tumor control was 83.5% for the whole group of patients, 94% for Group I, and 76.9% for Group II (p = 0.04). The early sequelae related to surgery or irradiation were self limited and disappeared by 6 months after the end of the treatment: ocular irritation (14 cases), scleral atrophy (5 cases), and neovascularization (7 cases). A greater incidence of sequelae was observed in Group II patients, but the difference between the groups was not statistically significant (p = 0.15). No significant correlation between treatment sequelae and treatment dose was noted: 29% sequelae with 30 Gy vs. 18.7% sequelae with 60 Gy (p = 0.32). No late complications have been observed. CONCLUSION: Adjuvant beta irradiation provides effective therapy for primary pterygium, is somewhate less effective in patients with recurrent pterygium, and is associated with a moderate rate of early and transient sequelae. PMID- 11020586 TI - Treatment-time-dependence models of early and delayed radiation injury in rat small intestine. AB - BACKGROUND: The present study modeled data from a large series of experiments originally designed to investigate the influence of time, dose, and fractionation on early and late pathologic endpoints in rat small intestine after localized irradiation. The objective was to obtain satisfactory descriptions of the regenerative response to injury together with the possible relationships between early and late endpoints. METHODS: Two- and 26-week pathologic radiation injury data in groups of Sprague-Dawley rats irradiated with 27 different fractionation schedules were modeled using the incomplete repair (IR) version of the linear quadratic model with or without various time correction models. The following time correction models were tested: (1) No time correction; (2) A simple exponential (SE) regenerative response beginning at an arbitrary time after starting treatment; and (3) A bi-exponential response with its commencement linked to accumulated cellular depletion and fraction size (the 'intelligent response model' [INTR]). Goodness of fit of the various models was assessed by correlating the predicted biological effective dose for each dose group with the observed radiation injury score. RESULTS: (1) The incomplete repair model without time correction did not provide a satisfactory description of either the 2- or 26 week data. (2) The models using SE time correction performed better, providing modest descriptions of the data. (3) The INTR model provided reasonable descriptions of both the 2- and 26-week data, confirming a treatment time dependence of both early and late pathological endpoints. (4) The most satisfactory descriptions of the data by the INTR model were obtained when the regenerative response was assumed to cease 2 weeks after irradiation rather than at the end of irradiation. A fraction-size-dependent delay of the regenerative response was also suggested in the best fitting models. (5) Late endpoints were associated with low-fractionation sensitivity and treatment-time dependence even in animal groups that exhibited minimal early mucosal reactions. CONCLUSION: Radiation injury scores in this rat small intestinal experimental model cannot be adequately described without time correction. 'Consequential' mechanisms contribute to the development of late effects, even in animals that do not develop severe early mucosal injuries. The initiation of the regenerative response is subject to a fraction-size-dependent mitotic delay and is linked to the level of accumulated cellular depletion. The response does not cease at the end of therapy but probably continues until maximal healing has taken place. PMID- 11020587 TI - Neointimal responses 3 months after (32)P beta-emitting stent placement. AB - PURPOSE: Studies have shown a potential benefit of brachytherapy in preventing restenosis. However, the effects of intravascular radiation on arterial healing have not been well-established. The purpose of this study was to explore the histologic changes following placement of beta-emitting radioactive stents in arteries focusing on intimal responses and endothelialization. METHODS AND MATERIALS: 3.0-mm beta-emitting (32)P stents (6-microCi and 24-microCi) were placed in rabbit iliac arteries with nonradioactive stents serving as controls. Animals were euthanized at 3 months and histologic assessment, morphometry, and analysis of endothelialization were performed. RESULTS: The lumen areas of 24 microCi stents (4.24 +/- 0.22 mm(2), p < 0.0007) and 6-microCi stents (4.23 +/- 0.49 mm(2), p < 0.01) were larger than control stents (3.64 +/- 0.44 mm(2)). The mean lumen percent stenosis was 11. 4 +/- 3.0% in the 24-microCi stents (p < 0.007 vs. 6-microCi stents and p < 0.0001 vs. control stents), 18.7 +/- 6.4% in the 6-microCi stents (p < 0.02 vs. control stents), and 25.0 +/- 4.9% in control stents. Neointimal area was least in the 24-microCi stent (54.2% smaller than controls and 42.7% smaller than 6-microCi); the neointimal area of the 6-microCi stents was 20.0% less than controls. The control stent neointima consisted of smooth muscle cells in a proteoglycan and collagen matrix. In contrast, the intima of radioactive stents showed persistent fibrin thrombus with nonconfluent areas of matrix. Actin-positive intimal cell density was reduced with radioactive stenting, but intimal cell proliferation was increased. Evans blue staining, an indicator of increased endothelial permeability, was present on 86 +/- 9% of the stented segment of 6-microCi stents vs. 10 +/- 11% in controls (p < 0.0001). Scanning electron microscopy demonstrated endothelialization of 97 +/- 8% of the intimal surface of control stents; in contrast, the midportion of the 6-microCi stents remained nonendothelialized, and only 33 +/- 15% (p < 0.0001) of the entire stent surface was endothelialized. CONCLUSIONS: (32)P beta-emitting stents reduce neointimal growth, but healing is incomplete with poor endothelialization at 3 months. Longer-term studies with complete arterial healing are needed to determine whether there is sustained neointimal inhibition by stent-delivered brachytherapy. PMID- 11020588 TI - Postimplant dosimetry for (125)I prostate implants: definitions and factors affecting outcome. AB - OBJECTIVE: An analysis of CT-based dosimetry was performed to assess the efficacy of the real time method of prostate implantation, explore the relationship of various dose descriptions and determine implant factors affecting outcome. METHODS AND MATERIALS: Between 7/95 and 8/99, 297 patients underwent (125)I implants for T1-T2 prostate cancer and had CT-based dosimetry performed (TG43 formalism). Dosimetry was performed 1 month postimplant. Using a dose-volume histogram, doses delivered to 100%, 95%, 90%, and 80% of the prostate (D100, D95, D90, D80, respectively) as well as percentages of the gland receiving 240 Gy, 160 Gy, 140 Gy (V240, V160, V140, respectively) were reported. Correlations between the various dose parameters and D90 were generated. The effect of the number of seeds implanted, seeds/volume, prostate volume, experience as assessed by time (8/01/99-date of implant), ultrasound probe (mechanical sector vs. dual phased electronic), and the ratio of the CT dosimetry prostate volume/ultrasound implant volume (CT/US vol) were analyzed. RESULTS: The median D100, D95, D90, and D80 values were 10,200 cGy, 15,655 cGy, 17,578 cGy, and 19,873 cGy, respectively. The median V240, V160, and V140 were 56%, 94%, and 98%, respectively. Correlations of dose descriptions found a close relationship of D95, D80, V240, V160, and V140 with D90 with r values of 0.928, 0.973, 0.911, 0.816, and 0.733, respectively. D100 correlated poorly with D90 (r = 0.099). Using a stepwise regression analysis, CT/US vol ratio, prostate volume, and seed number were the only significant factors affecting D90 with CT/US vol ratio having the greatest effect. The dual-phased electronic probe was associated with fewer D90 values of less than 140 Gy (2%) compared to the mechanical sector probe (14%) (p = 0.02). CONCLUSION: CT-based dosimetry results reveal the real-time implant technique to be an effective method of prostate implantation. Factors associated with more precise implantation, such as decreased postimplant edema, new technology, and increased number of seeds will lead to higher D90 values. PMID- 11020589 TI - Receipt of preventive health care services by lesbians. AB - BACKGROUND: We measured receipt of age-appropriate preventive health services by lesbians and assessed whether provider and individual characteristics, including disclosure of sexual orientation, are independently associated with receipt of these services. METHODS: A questionnaire was printed in a national biweekly gay, lesbian, and bisexual news magazine, and self-identified lesbians living in all U. S. states (N =6935) responded to the survey. Main outcome variables were receipt of a Pap smear within the preceding 1 and 2 years and, for women aged > or= 50, receipt of a mammogram within the past 1 and 2 years. RESULTS: Fifty-four percent had Pap smears within 1 year and 71% within 2 years, with increasing rates among older and more educated respondents. Seventy percent of respondents aged > or = 50 had a mammogram in the past year, and 83% within 2 years; rates did not vary significantly controlling for education. Sixty percent had disclosed their sexual orientation to their regular health care provider. Controlling for patient and provider characteristics, disclosure was independently associated with receipt of Pap smears, but not mammograms. CONCLUSIONS: It is important for providers to identify their lesbian patients' unmet needs for preventive health care. Additionally, it is important for providers to provide complete and appropriate preventive health care for their lesbian patients. Further research is needed to determine why lesbians are not receiving Pap smears at the recommended rate and whether this disparity is reflective of aspects of cervical cancer screening or indicates a more general problem with access to health care including receipt of preventive services. PMID- 11020590 TI - Colorectal cancer screening participation by older women. AB - OBJECTIVE: Although recent screening guidelines recommend annual fecal occult blood testing (FOBT) for adults aged > or = 50, a number of studies report that these tests are underused. Systematic efforts to increase awareness of colorectal cancer (CRC) and to promote screening participation are needed to meet national objectives for CRC control. METHODS: This study examined CRC-screening practices and evaluated factors related to recent participation in screening by FOBT in a sample of women aged 50 to 80 who were surveyed about their use of clinical preventive services at Group Health Cooperative, a managed care organization in western Washington State. RESULTS: Of the 931 women eligible for analysis, 75% reported ever having been screened by FOBT and 48% reported having been screened within 2 years before the survey. Participation in screening did not vary by demographic characteristics or by perceived or actual risk of CRC. Women with a positive attitudes toward CRC screening had sevenfold greater odds of recent screening by FOBT (odds ratio=7.1; 95% confidence interval, 4.4 to 11.6). Only 58% of study women reported that their physicians encouraged CRC screening, but this factor was strongly related to participation (odds ratio=12.7; 95% confidence interval, 6.6 to 24.4). CONCLUSIONS: We identified several areas in which understanding of CRC risk may be low. As a whole, these findings suggest that effective strategies to control CRC may include efforts to improve knowledge of risk and prevention, but must also appeal directly to primary care physicians to identify and address their barriers to screening recommendations. PMID- 11020591 TI - Mortality rates and causes among U.S. physicians. AB - CONTENT/OBJECTIVES: No recent national studies have been published on age at death and causes of death for U.S. physicians, and previous studies have had sampling limitations. Physician morbidity and mortality are of interest for several reasons, including the fact that physicians' personal health habits may affect their patient counseling practices. METHODS: Data in this report are from the National Occupational Mortality Surveillance database and are derived from deaths occurring in 28 states between 1984 and 1995. Occupation is coded according to the U.S. Bureau of the Census classification system, and cause of death is coded according to the ninth revision of the International Classification of Diseases. RESULTS: Among both U.S. white and black men, physicians were, on average, older when they died, (73.0 years for white and 68.7 for black) than were lawyers (72.3 and 62.0), all examined professionals (70.9 and 65.3), and all men (70.3 and 63.6). The top ten causes of death for white male physicians were essentially the same as those of the general population, although they were more likely to die from cerebrovascular disease, accidents, and suicide, and less likely to die from chronic obstructive pulmonary disease, pneumonia/influenza, or liver disease than were other professional white men. CONCLUSIONS: These findings should help to erase the myth of the unhealthy doctor. At least for men, mortality outcomes suggest that physicians make healthy personal choices. PMID- 11020592 TI - Point-of-entry screening for C. trachomatis In female army recruits. Who derives the cost savings? AB - BACKGROUND: Screening women for genital Chlamydia trachomatis at entry to military service provides an opportunity to reduce costs associated with sequelae of this infection. However, financial responsibility for screening may be debated. More than 50% of recruits return to civilian life within 2 years. The military and the civilian health care systems would both benefit from a screening program. OBJECTIVE: To assess the cost-effectiveness and relative cost savings to the military and civilian health sectors of three screening strategies for U.S. Army female recruits for C. trachomatis using urine ligase chain reaction: screening all recruits, screening recruits aged < or = 25 years, and no screening. METHODS: We applied a decision analytic model. Cost factors included screening, lost military training, morbid pelvic inflammatory disease, and other sequelae. Using a 5-year analytic horizon, we conducted analyses from military and civilian perspectives. RESULTS: Screening 10,000 female army recruits would cost 193,500 dollars and prevent 282 cases of sequelae, with a projected savings of 53,325 dollars to the military and 505,053 dollars to the civilian sector. From a military perspective, screening women aged < or = 25 years provided the highest cost savings. Screening all female recruits incurred an incremental cost of $1199 per sequela prevented. From a civilian perspective, screening all recruits offered the greatest cost savings. CONCLUSIONS: Screening female Army recruits for C. trachomatis offers substantial savings in health care costs for both the military and civilian health care systems. Relative financial benefit derived from recruit screening is disproportionate; greatest cost savings are enjoyed by the civilian sector. PMID- 11020593 TI - Misconceptions about STD-protective behavior. Project RESPECT Study Group. AB - CONTEXT: Misconceptions about STD-protective behaviors have not been studied before and after STD counseling. Further, to the best of our knowledge, the relationship of these misconceptions to condom use and STD incidence has not previously been described in published reports. OBJECTIVES: The main purpose of the study was to determine the prevalence of misconceptions about STD prevention among STD clinic attendees (N=3498) in five large cities, as well as whether misconceptions decreased after STD diagnosis, STD counseling, or both. The study also identified predictors of persistent misconceptions and determined the relationship of STD incidence and unprotected sex to persistent misconceptions. METHODS: Data from a randomized controlled trial evaluating HIV/STD counseling interventions (Project RESPECT) were used for the present analyses. Participants completed an interview upon study enrollment and every 3 months following enrollment for a 1-year period. A portion of the interview assessed participants' misconceptions about STD-protective behaviors. RESULTS: At baseline, 16.3% believed that washing the genitals after sex protected from STDs. Likewise, urinating after sex (38.7%), douching (45.7%), and use of oral contraceptives (19. 9%) were believed to prevent STDs. Prevalence of misconceptions was significantly diminished at a 3-month follow-up (p<.001). Those continuing to have misconceptions were more likely to be aged > or = 24 and African American. Those continuing to have these misconceptions did not have higher STD incidence. CONCLUSIONS: Misconceptions about STD-protective behaviors are common, and the event of an STD or STD counseling or both generally reduces these misconceptions. Although these misconceptions may not directly translate into risky behavior, they may preclude movement toward safer sex. PMID- 11020594 TI - Factors affecting environmental awareness among Head Start families in Mississippi. AB - BACKGROUND: Socioeconomic and racial/ethnic disparities in health status in the United States may be attributed in part to environmental injustice and differential exposure to environmental hazards among low-income and/or minority populations. However, the environmental justice movement has historically focused on equity in the siting of point-source polluting facilities, giving little attention to environmental hazards and environmental awareness at the level of the individual household. METHODS: Heads of 763 low-income households participating in Head Start programs in 20 counties of the Mississippi Delta region were surveyed regarding their education, the physical environment of their home and workplace, sources of food and water, awareness of local polluting sites/facilities, knowledge of government agencies, and behaviors that may affect their health or impact their local environment. Survey results were compared to demographic, socioeconomic, and environmental quality indicators. RESULTS: Significant associations existed between both education and race/ethnicity and the responses of survey participants. Being African American was more commonly associated with poor quality-of-life indicators such as renting substandard older homes and living in close proximity to areas of unfavorable watershed quality. Higher education was more commonly and positively associated with indicators of heightened environmental awareness and increased political empowerment. No association was observed between race/ethnicity and the prevalence of polluting facilities. However, a significant association existed between race/ethnicity and indicators of environmental quality/integrity. CONCLUSIONS: Environmental health education interventions that target individual households may be a useful mechanism for increasing the access of low-income communities to government health resources and reducing adverse health effects from the environment. However, racial/ethnic disparities in education and health remain an important consideration. PMID- 11020595 TI - State youth-access tobacco control policies and youth smoking behavior in the United States. AB - CONTEXT: State policies aimed at controlling youth access to tobacco are an important component of public health efforts to reduce smoking prevalence among youth and prevent subsequent disease. OBJECTIVES: This study sought to assess the extensiveness of state youth access tobacco control legislation in the United States, describe how state policies changed over a 4-year period, explore how various political and economic characteristics are related to state policies, and determine the relationship of youth smoking behavior to state youth tobacco control policies. DESIGN: This descriptive and correlational study utilized data from multiple national surveillance, economic, and sociodemographic data sets. PARTICIPANTS: All 50 states and the District of Columbia provided economic and political data. A standard tobacco-control policy score was developed by an expert panel for each state. Aggregated state-level measures of youth smoking behavior were provided by 79,491 youth in 33 states and the District of Columbia. MAIN OUTCOME MEASURES: Included were extent of state tobacco control policies and changes over time, relationship between state political and economic characteristics and tobacco control policies, and relationship between state policies and youth smoking behavior. RESULTS: State policy scores increased in variability and in mean value over the 4-year period, from a mean score of 7.2 in 1993 to 9.0 in 1996. State policy scores were significantly correlated with several state political and economic variables. States with more extensive tobacco control policies had significantly lower youth smoking rates. There was some evidence that a strong state tobacco economy may limit the effectiveness of tobacco control policies on youth smoking rates. CONCLUSIONS: It is possible to reliably measure the extent to which states are achieving important public health goals in limiting youth access to tobacco products. Comprehensive state tobacco control policies are important for increasing prevention and cessation of smoking among youth. PMID- 11020596 TI - Restrictions on smoking at home and urinary cotinine levels among children with asthma. AB - OBJECTIVES: The purpose of this study was to determine the extent to which various levels of restrictions on smoking in the home may be associated with children's exposure to environmental tobacco smoke (ETS). METHODS: The methodology consisted of a cross-sectional survey involving 249 children with asthma aged 1 to 11 attending hospital outpatient clinics, with at least one parent who smoked, linked to the child's urinary cotinine to creatinine ratios (CCR). RESULTS: After adjustment for child's age, mother's smoking status, and total parental daily cigarette consumption, a total ban was associated with significantly lower urinary CCR levels (7.6 nmol/mmol) than bans with exceptions or limited smoking in the home. Where exceptions to bans were made (14.9 nmol/mmol), children's urinary CCR levels were no different from homes in which smoking was allowed in rooms the child rarely frequented (14.1 nmol/mmol). These two intermediate levels of restriction were in turn associated with significantly lower CCR levels than unrestricted smoking in the home (26.0 nmol/mmol). CONCLUSIONS: Making exceptions to bans on smoking at home measurably undermines the protective effect of a ban. However, making some exceptions to a ban and limiting smoking to rooms where the child rarely goes may result in reduced exposure to ETS, compared with unrestricted smoking. PMID- 11020597 TI - Correlates of postpartum smoking relapse. Results from the Pregnancy Risk Assessment Monitoring System (PRAMS). AB - BACKGROUND: Using population-based data from the Pregnancy Risk Assessment Monitoring System (PRAMS), this study examines the prevalence and intensity of smoking before, during, and after pregnancy, and identifies correlates of postpartum smoking relapse. METHODS: Women who delivered live births in 1996 responded to a mailed questionnaire approximately 2 to 6 months after delivery (N =17,378). Data from 10 states participating in PRAMS were included in the study, and the overall participation rate was 75%. Analyses were adjusted for survey design and sampling strategy. Logistic regression analysis identified independent correlates of smoking relapse. RESULTS: Overall, 25.6% of women reported cigarette smoking before pregnancy. Among women who smoked before pregnancy, 44.5% quit during pregnancy. Among women who quit during pregnancy, half relapsed by the time of the survey. Independent correlates associated with increased risk of postpartum relapse included African American race/ethnicity, multiparity, high maternal weight gain, late or no prenatal care, and stressful life events. CONCLUSIONS: Correlates of postpartum smoking relapse identified by this study may contribute to the development of effective and targeted interventions to maintain long-term smoking cessation. PMID- 11020598 TI - A curriculum for environmental genetics education. AB - INTRODUCTION: Environmental genetics is a scientific area concerned with interactions between genes and the environment. Progress in this field, coupled with the growth in genetic testing, has great potential for improving human health. There are also ethical, legal, and social concerns surrounding advances in environmental genetics and genetic testing. Because genetic information is rapidly increasing in our society, the public needs to learn more about scientific progress and policy issues in these areas. OBJECTIVE: To describe a curriculum for the public on environmental genetics and genetic testing. PROGRAM: In 1998, the Department of Environmental Health (Center for Environmental Genetics), University of Cincinnati, began an outreach project for the public called Learning Exchange for Genetic and Environmental Disease Solutions (LEGENDS). The project fosters awareness and understanding of environmental genetics and genetic testing with discussion of related policy issues. The curriculum includes brief lectures and discussions based on thematic modules and a set of interactive exercises to be conducted in small groups. More than 100 persons have attended instructional sessions sponsored by LEGENDS at the time of this writing. SIGNIFICANCE: The curriculum appears to be a potentially useful resource for educating the public about environmental genetics, genetic testing, and related policy issues. This project has implications for other organizations working to further genetics education. PMID- 11020599 TI - Enhancing patient safety and reducing medical error. The role of preventive medicine. PMID- 11020600 TI - Elegy for fifty-six black female physicians. PMID- 11020601 TI - Notes from the Association of Teachers of Preventive Medicine. Affirmative action: it's everybody's business. PMID- 11020602 TI - Searching for the perfect phakic intraocular lens. PMID- 11020603 TI - New pieces for the puzzle: nonsteroidal anti-inflammatory drugs and corneal ulcers. PMID- 11020605 TI - Consultation section PMID- 11020604 TI - Consultation section. Cataract surgical problem. PMID- 11020607 TI - Consultation section PMID- 11020606 TI - Consultation section PMID- 11020608 TI - Consultation section PMID- 11020610 TI - Consultation section PMID- 11020609 TI - Consultation section PMID- 11020611 TI - Flexible iris hooks for phacoemulsification in patients with iridoschisis. AB - Flexible iris hooks, or retractors, can be used to facilitate cataract removal by phacoemulsification in patients with primary iridoschisis. This rare condition is associated with fibrillary degeneration of the iris, narrow drainage angles, and cataract. In addition to their conventional use as iris retractors, iris hooks can control the degenerate fibrillary iris stroma to improve the view and access to the lens, preventing further damage during phacoemulsification and cortical cleanup. Iris hooks are widely available, easily handled, and can transform a difficult case into one that is almost routine. PMID- 11020612 TI - Quarters extraction technique for manual phacofragmentation(1). AB - In the quarters extraction technique, the nucleus is manually split and the fragments then removed. A 5.5 to 6.5 mm sclerocorneal single-plane incision is made. After capsulorhexis, hydrodissection, hydrodelineation, and surface cortex aspiration, the edge of the nucleus is prolapsed into the anterior chamber. The front quarter of the nucleus is cut and removed with a nucleus puncher. A corner of the remaining three quarters of nucleus is wedged into the wound and rotated out with a claw vectis. Among the initial 120 cases, there were no posterior capsule ruptures, and the mean endothelial cell loss at 3 months was 8.7% +/- 6.5% (SD). Because there is no need to deeply insert instruments at the time of nuclear fragmentation, this technique can be performed safely and easily in most cases except in eyes with very large nuclei. PMID- 11020613 TI - Surgical correction of severe myopia with an angle-supported phakic intraocular lens. AB - PURPOSE: To evaluate the effectiveness, predictability, and safety of a fourth generation angle-supported anterior chamber phakic intraocular lens (IOL) in patients with severe myopia. SETTING: Refractive Surgery and Cornea Unit, Alicante Institute of Ophthalmology, Miguel Hernandez University School of Medicine, Alicante, Spain. METHODS: In this prospective study, 23 eyes (16 patients) with a mean preoperative myopia of -19.56 diopters (D) +/- 1.76 (SD) (range -16.75 to -23.25 D) were implanted with the ZSAL-4 phakic IOL. Basic examinations were performed before and after surgery in all patients. Eighteen eyes were also studied by clinical specular microscopy, and the endothelium was analyzed for cell density, coefficient of variation in cell size, and hexagonality. The location of the IOL in the anterior chamber was evaluated in 10 eyes by A-scan biometry. To evaluate haptic geometry, a theoretical mechanical model was used. The follow-up was 24 months in all patients. RESULTS: Uncorrected visual acuity was 20/40 or better in 13 eyes (56.0%) 12 months after surgery and in 14 eyes (60.8%) at 24 months. Best spectacle-corrected visual acuity improved 0.19 at 12 and 24 months (0.1 = 1 line) from preoperative values. The efficacy index was 1.12 at 24 months and the safety index, 1.45. The mean postoperative spherical equivalent was -0.65 +/- 0.65 D at 24 months. The postoperative spherical equivalent was within +/-1.00 D of emmetropia in 19 eyes (82.6%) at 12 and 24 months. The mean endothelial cell loss was 3.50% at 12 months and 4.18% at 24 months. The coefficient of variation in cell size decreased from 0.34 before surgery to 0.28 at 24 months after surgery. The IOL was located 0.79 +/- 0.24 mm in front of the crystalline lens. Postoperative complications included night halos in 6 eyes (26.1%) at 12 and 24 months and pupil ovalization in 4 eyes (17.4%). Intraocular lens rotation was observed in 10 eyes (43.5%) at 24 months. Two eyes (8. 7%) developed a slight inflammatory response during the first 6 months. Our mechanical model predicted that the compression forces against the angle structures were greater at the first footplate than at the second. CONCLUSIONS: Implantation of the ZSAL-4 IOL in the anterior chamber of phakic eyes was effective and predictable in correcting severe myopia. However, the ZSAL 4 did not prevent pupil ovalization, IOL rotation, or low-grade postoperative uveitis. Improvements in haptic design following our mechanical model could decrease these haptic-related complications. PMID- 11020614 TI - Cataract surgery in highly myopic eyes corrected by phakic anterior chamber angle supported lenses(1). AB - PURPOSE: To assess the incidence of cataract, potential causes of its development, and the outcome of cataract surgery after previous implantation of phakic angle-supported anterior chamber intraocular lenses (AC IOLs) in highly myopic patients. SETTING: Instituto Oftalmologico de Alicante, Department of Refractive Surgery, Spain. METHODS: Two hundred sixty-three highly myopic phakic eyes of 160 patients had implantation of a phakic AC IOL. Follow-up was up to 8 years (range 38.4 to 103.2 months). Eyes that subsequently developed cataract had cataract extraction and were studied during follow-up for clinical association to other preoperative or postoperative data. Cataract surgery was performed after phakic IOL explantation by phacoemulsification and posterior chamber IOL (PC IOL) implantation (Domilens-Chiron AL3). RESULTS: Nuclear cataract developed in 9 cases (3.42%) a mean of 42.91 months +/- 17.7 (SD) after phakic AC IOL implantation. Final best spectacle-corrected visual acuity (BSCVA) was not significantly different from that after phakic AC IOL implantation (P =.25, paired Student t test). Mean endothelial cell loss after cataract surgery was 6.87% +/- 0.42% cells/mm(2). Age at implantation of older than 40 years and axial length greater than 30. 0 mm were the 2 factors significantly related to nuclear cataract development (r = 1.69 and 1.98, respectively; P <.05). CONCLUSIONS: There is a potential risk of nuclear cataract development after phakic AC IOL implantation to correct high myopia in patients older than 40 years and with very high (greater than 30.0 mm) axial myopia. Phakic IOL explantation, phacoemulsification, and PC IOL implantation successfully resolved this complication. The benefits in terms of BSCVA and spherical equivalent obtained after phakic AC IOL implantation were preserved after cataract surgery. PMID- 11020615 TI - Diffuse lamellar keratitis associated with epithelial defects after laser in situ keratomileusis. AB - PURPOSE: To describe the association between the presence of epithelial defects and the development of diffuse lamellar keratitis (DLK), "Sands of the Sahara" syndrome, following laser in situ keratomileusis (LASIK). SETTING: Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, USA. METHODS: In this retrospective study, the postoperative clinical course and surgical outcomes in 735 eyes of 358 consecutive patients who had myopic LASIK between December 1998 and August 1999 were reviewed. Of the 735 procedures, 680 were primary LASIK procedures and 55 were retreatments performed by lifting the existing flaps. The incidence and severity of DLK and the relationship of DLK to epithelial defects were tabulated. RESULTS: Diffuse lamellar keratitis developed in 9 of the 16 eyes that had postoperative epithelial defects. Seventeen eyes without epithelial defects also developed DLK. The presence of an epithelial defect increased an individual's risk of developing DLK 24 times (95% confidence interval, 13 to 45). In 8 eyes, the inflammation resolved following treatment with intense topical corticosteroids. One eye had irrigation under the flap because of dense central inflammation; the final outcome was mild inferior corneal steepening. All eyes recovered preoperative best spectacle-corrected visual acuities. CONCLUSION: Patients who have epithelial defects of any size following LASIK are at significantly increased risk of developing DLK. PMID- 11020617 TI - Comparison of tear secretion and tear film instability after photorefractive keratectomy and laser in situ keratomileusis. AB - PURPOSE: To evaluate and compare tear secretion and tear film instability following photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). SETTING: Department of Ophthalmology, Yonsei University School of Medicine, Seoul, Korea. METHODS: In a prospective study, 36 eyes (21 patients) had PRK and 39 eyes (25 patients) had LASIK to correct myopia. Tear secretion and tear film instability were tested preoperatively and 3 and 6 months postoperatively using Schirmer test values, tear breakup time (BUT) scores, and tear osmolarity. RESULTS: Six months after surgery, the change in Schirmer test values from preoperative levels was -14.57% +/- 6.39% (SD) in the PRK eyes and 23.40% +/- 5.94% in the LASIK eyes and the change in BUT scores, -12.54% +/- 8.28% and -18.79% +/- 13.01%, respectively. The change in tear osmolarity was 14.95% +/- 6.46% and 35.63% +/- 8.51%, respectively. CONCLUSIONS: The decrease in tear secretion was greater after LASIK than after PRK at 6 months. Proper treatment of dry eye is required after LASIK and PRK, particularly in the LASIK postoperative period. PMID- 11020616 TI - Anatomic factors affecting microkeratome placement in laser in situ keratomileusis(2)(2). AB - PURPOSE: To identify the significant anatomic factors that affect placement of 2 microkeratomes and compare the ease of their placement. SETTING: The Laser Center, Houston, Texas, USA. METHODS: Anatomic factors affecting microkeratome placement were evaluated in 250 eyes of 137 myopic patients in a prospective consecutive-enrollment study. Anatomic factors measured included vertical orbital width, horizontal interpalpebral fissure (HPF), corneal diameter, and position of the corneal apex relative to the lateral orbital rim (PCL). Vertical orbital width and HPF were measured with a micrometer caliper (Mitutoyo); corneal diameter, with Orbscan topography (Bausch & Lomb Surgical); and PCL, with a Hertel exophthalmometer. Patients were randomly divided into 2 groups (125 eyes/group) to have surgery with the Automated Corneal Shaper((R)) (ACS) (Bausch & Lomb Surgical) or Hansatome (Bausch & Lomb Surgical) microkeratome. Microkeratome placement was graded by 1 surgeon as "easy" (grades 1, 2, and 3) or "difficult" (grades 4 and 5). Results were correlated with anatomic factors to evaluate possible anatomic influences on the difficulty of microkeratome placement. RESULTS: The PCL and vertical orbital width were statistically significant factors in predicting the difficulty of ACS microkeratome placement (P <.05). For the Hansatome microkeratome, since only 1 eye was graded as a difficult placement, the anatomic parameters affecting placement could not be evaluated. There were no statistically significant between-group differences in mean age and means of all anatomic factors measured (independent t test, P >.05). Difficult applanations were encountered in the ACS group (8/125 eyes, 6.4%) more than in the Hansatome group (1/125 eyes, 0.8%), which was a statistically significant difference by the Pearson chi-square test (P <.05). CONCLUSIONS: The PCL and vertical orbital width were significant anatomic factors affecting placement of the ACS microkeratome. The anatomic factors affecting Hansatome microkeratome placement could not be determined. The percentage of difficult placements was significantly lower with the Hansatome than with the ACS. PMID- 11020618 TI - Goldmann applanation tonometry after intacs corneal ring segments(1). AB - PURPOSE: To evaluate the change of intraocular pressure (IOP) after implantation of Intacs (KeraVision) corneal ring segments using Goldmann applanation tonometry (GAT). SETTING: Multicenter clinical trial for U.S. Food and Drug Administration application conducted at 10 U.S. sites. METHODS: One-year follow-up data from a phase III clinical trial (n = 359) were reviewed. Intraocular pressure was measured by GAT preoperatively and 1 week and 1, 3, 6, and 12 months after Intacs implantation. The untreated fellow eyes were controls. Change from baseline (preoperative) IOP was calculated and tested for correlation with age and change from baseline in mean keratometry, manifest refraction spherical equivalent (MRSE), and pachymetry (central corneal thickness). Mean IOP in treated and control groups was compared through month 6. RESULTS: At all postoperative examinations, mean IOP in the Intacs eyes was significantly lower (-0.39 to -1.75 mm Hg; all P 0.10 g); 8 of these developed glaucoma. The other 6 eyes had no or minimal after-cataract and did not develop secondary glaucoma. The relationship between after-cataract and secondary glaucoma was statistically significant. CONCLUSION: A significant relationship between the amount of after-cataract and the development of secondary glaucoma was found in aphakic infant rabbit eyes. PMID- 11020627 TI - Corneal endothelial toxicity of different lidocaine concentrations. AB - PURPOSE: To examine the potential damaging effect on the corneal endothelium of unpreserved lidocaine in concentrations of 1%, 5%, and 10%. SETTINGS: Department of Ophthalmology, Charite Medical Faculty, Humboldt University, Berlin, Germany. METHODS: Experimental porcine corneas (n = 18) were exposed to 100 microL of unpreserved lidocaine hydrochloride at concentrations of 1%, 5%, and 10% for 60 minutes. Additional corneas (n = 6) were treated with lidocaine hydrochloride 1% for 30 minutes to simulate clinical conditions. Balanced salt solution (BSS((R))) served as a control to evaluate corneal endothelial cell damage using Janus Green photometry. Morphology, damage pattern, and changes in the ultrastructural appearance of corneal endothelial cells were examined by light and scanning electron microscopy. RESULTS: Lidocaine 1% used for 30 or 60 minutes did not cause significantly more corneal endothelial damage (mean 3.00% +/- 0.76% [SD] and 3.26% +/- 1.00%, respectively) than in the control group (mean 3.32% +/- 0. 86%) (P >.01). Significant corneal endothelial cell loss was observed with lidocaine 5% (mean 10.7% +/- 6.4%) (P <.001) and lidocaine 10% (42.3% +/- 17.0%) (P <.001). CONCLUSION: Experimental exposure of corneal endothelial cells to higher concentrations of lidocaine resulted in significant cell loss, indicating that the 1% concentration only should be used clinically. PMID- 11020629 TI - Topography changes associated with sublamellar epithelial ingrowth after laser in situ keratomileusis. AB - Laser in situ keratomileusis (LASIK) was performed in 1 eye of a patient for correction of myopia. The patient was evaluated postoperatively using both Placido-disk-based videokeratography and rasterstereography. The patient developed an epithelial defect and subsequent sublamellar epithelial ingrowth after LASIK. The Placido-disk system demonstrated an area of flattening over the area of epithelial ingrowth, and rasterstereography more specifically identified the area over the epithelial ingrowth as an area of relative elevation. PMID- 11020628 TI - Interaction between inflammatory cells and heparin-surface-modified intraocular lens. AB - PURPOSE: To investigate the interaction and adherence of inflammatory cells to a heparin-surface-modified intraocular lens (HSM IOL). SETTING: Department of Ophthalmology, Tokyo Medical University Hospital, Tokyo, Japan. METHODS: Splenic mononuclear leukocytes from rats with experimental autoimmune uveitis were cultured with the optic of an HSM IOL for 96 hours. The number of adherent cells on the HSM IOL surface was measured with and without the addition of interphotoreceptor retinoid-binding protein and concanavalin A (ConA) to the culture medium. The adherent cells were observed under a light microscope or a scanning electron microscope. RESULTS: Interphotoreceptor retinoid-binding protein and ConA increased the number of adherent cells on the HSM IOL relative to the control. Adherent cells on the HSM IOL were small and round, considered to be mainly lymphocytes. CONCLUSION: Activated lymphocytes tended to adhere to the surface of the HSM IOL. PMID- 11020630 TI - Complete capsular bag distension syndrome. AB - After conventional extracapsular cataract extraction and in-the-bag intr aocular lens (IOL) implantation 3 years previously, the partially cut su perior one-third of the anterior capsulotomy flap resealed in the eye of a 55-year-old woman. With the integrity of the capsular bag restored, it distended fully; the IOL was in the center and milky fluid accumulated inside. A neodymium:YAG laser was used to drain the milky fluid and di srupt the opacified anterior and posterior capsules, restoring vision. PMID- 11020631 TI - Corneal laceration with total but isolated aniridia caused by a pecking injury. AB - A 38-year-old man sustained a left eye injury after being attacked by a mynah bird. Ocular examination revealed a beak-shaped, full-thickness corneal laceration with total aniridia in the left eye. No other ocular injury was observed, and the lens and posterior segment remained normal over the subsequent 3 months. A penetrating keratoplasty was planned for visual restoration. PMID- 11020632 TI - Posterior capsule rupture after a paint-pellet injury. AB - We report a case in which a high-velocity paint pellet injury to the eye ruptured the posterior lens capsule. In the setting of blunt trauma, posterior capsule rupture has rarely been reported. Recognizing such a rupture allows the surgeon to plan the best surgical approach. PMID- 11020633 TI - Peripheral ulcerative keratitis after clear corneal cataract extraction(1). AB - A previously healthy 80-year-old man had uneventful clear corneal cataract extraction. An extensive peripheral corneal infiltrate with overlying epithelial defect at the incision site was noted at the regular follow-up visit 1 week after surgery. Corneal cultures showed no evidence of infectious keratitis. A systemic evaluation uncovered early-stage, active rheumatoid arthritis. This case illustrates that peripheral ulcerative keratitis may occur with a small clear corneal incision and may be the presenting sign of a previously undiagnosed rheumatoid disease. PMID- 11020634 TI - Posterior dislocation of a plate-haptic silicone intraocular lens with large fixation holes. AB - We present a patient who had uneventful phacoemulsification with in-the- bag implantation of a silicone plate-haptic posterior chamber intraocula r lens (IOL) with 1.15 mm fixation holes. Six months after a neodymium: YAG laser capsulotomy, the IOL spontaneously dislocated posteriorly. A pars plana vitrectomy was performed, the IOL was explanted, and a 3-piece loop-haptic IOL was implanted in the ciliary sulcus. Eight weeks after surgery, best corrected visual acuity was 20/20. PMID- 11020635 TI - Sleep-related disorders in neurologic disease during childhood. AB - Sleep disorders commonly are associated with neurologic disorders in childhood. This review discusses primary sleep disorders that affect children with primary neurologic diseases. Primary sleep disorders are discussed as they relate to the primary neurologic disease. In addition, sleep disorders secondary to neurologic disorders commonly seen in the practice of pediatric neurology are reviewed. A useful sleep history to improve diagnostic and therapeutic interventions is outlined. PMID- 11020636 TI - Predictive value of proton magnetic resonance spectroscopy in pediatric closed head injury. AB - We studied 26 infants (1-18 months old) and 27 children (18 months or older) with acute nonaccidental (n = 21) or other forms (n = 32) of traumatic brain injury using clinical rating scales, a 15-point MRI scoring system, and occipital gray matter short-echo proton MRS. We compared the differences between the acutely determined variables (metabolite ratios and the presence of lactate) and 6- to 12 month outcomes. The metabolite ratios were abnormal (lower NAA/Cre or NAA/Cho; higher Cho/Cre) in patients with a poor outcome. Lactate was evident in 91% of infants and 80% of children with poor outcomes; none of the patients with a good outcome had lactate. At best, the clinical variables alone predicted the outcome in 77% of infants and 86% of children, and lactate alone predicted the outcome in 96% of infants and 96% of children. No further improvement in outcome prediction was observed when the lactate variable was combined with MRI ratios or clinical variables. The findings of spectral sampling in areas of brain not directly injured reflected the effects of global metabolic changes. Proton MRS provides objective data early after traumatic brain injury that can improve the ability to predict long-term neurologic outcome. PMID- 11020637 TI - Linear hyperechogenicity within the basal ganglia and thalamus of preterm infants. AB - Linear hyperechogenicity (LHE) within the basal ganglia and thalamus is an uncommon sonographic finding in preterm infants and is of unclear significance. The study objectives were to determine the clinical characteristics and neurodevelopmental outcome in preterm infants who develop LHE. Ten preterm and 20 control infants were evaluated developmentally at 18 months adjusted age using the Bayley Scales of Infant Development. LHE was diagnosed at 4 weeks (range = 1 11). Antenatal glucocorticoid therapy was more common in infants with LHE than in the control infants (90% vs 45%). Four (44%) of nine LHE infants and no control infants were positive for cytomegalovirus (P = 0.02, and three of 10 LHE infants and no control infants had a hypothyroid (P = 0.03). The mental development scores and behavioral evaluation results were lower in the infants with LHE than in the control infants (73.7 +/- 9.7 vs 83.7 +/- 9.4, P = 0.01 and 23.7 +/- 20.1 vs 43.9 +/- 25.4, P = 0.04, respectively). The infants without LHE also had poorer motor quality (22.8 +/- 20. 5 vs 55.7 +/- 37.4, P = 0.02) and lower emotional regulation scores (25.7 +/- 16 vs 42.3 +/- 24, P = 0.06) than the control infants. Preterm infants with LHE are at an increased risk of adverse neurodevelopmental outcome and, in particular, cognitive and behavioral performance. The sonographic evolution of LHE may be a marker of a diffuse insult to the brain. PMID- 11020638 TI - Alternating hemiplegia of childhood: clinical manifestations and long-term outcome. AB - We present our analysis of 44 patients with alternating hemiplegia of childhood. The clinical course usually consisted of three phases. The first was dominated by abnormal eye movements and dystonic episodes, the second by hemiplegic spells and psychomotor regression, and the third by persistent developmental delay and fixed neurologic deficits. The age of onset was 0-54 months (mean = 7.9 +/- 13 months). The presenting signs included abnormal ocular movements in 65%, dystonia in 60%, and hemiplegia in 32%. Patients with an early onset of the disorder and an early appearance of hemiplegic spells faired the poorest developmentally. Developmental delay was present in 91%, ataxia in 68%, choreoathetosis in 50%, and seizures in 18%. Laboratory investigations suggested mitochondrial abnormalities and cerebrovascular dysfunction in several patients. Numerous therapies were largely ineffective. Flunarizine reduced the duration, severity, and frequency of the hemiplegic attacks in 78%. Patients who received flunarizine did not differ developmentally from those who did not. Our data suggest that flunarizine does not adversely affect and may favorably influence the outcome in patients with alternating hemiplegia of childhood. Additionally, the occurrence of autosomal dominant cases of the syndrome, although rare, suggests that, in addition to mitochondrial dysfunction, genetic factors may be important. PMID- 11020639 TI - Carbamazepine and valproic acid: effects on the serum lipids and liver functions in children. AB - We aimed to determine the effects of carbamazepine, which induces liver microsomal enzymes, and valproic acid on the serum lipids and liver function test results in epileptic children. Thirty-eight epileptic children (18 males, 20 females, mean age 8.6 +/- 3.9 years) were evaluated for serum lipids and liver function test results at the onset and the second and sixth months of antiepileptic therapy. The results of the children receiving carbamazepine (n = 31) and valproic acid (n = 7) were compared. In addition, the values obtained at different periods of treatment were compared within each group. The differences in the serum lipid levels and liver function test results of the children in the carbamazepine group and the valproic acid group were not statistically significant throughout the study. However, the total cholesterol, low-density lipoprotein, total cholesterol/high-density lipoprotein, and gamma glutamyl transferase levels were significantly increased in the carbamazepine group during treatment (P < 0.05) but not in the valproic acid group. Carbamazepine treatment alters the serum lipid profile of the children in such a way that it facilitates the development of atherosclerosis. Valproic acid does not alter the levels of the serum lipids. PMID- 11020640 TI - Successful treatment of rheumatic chorea with carbamazepine. AB - Carbamazepine has been used successfully in the treatment of different movement disorders and was recently reported to be effective for nonhereditary chorea. In view of the significant side effects associated with the drugs currently used to treat chorea, we sought to further evaluate the efficacy of carbamazepine in children with rheumatic chorea. The study was prospective and included 10 children with chorea (eight females and two males; age range = 7-16 years) referred to our Pediatric Rheumatology Clinic between 1995 and 1999. Nine had rheumatic fever and one had antiphospholipid antibody syndrome that later evolved to systemic lupus erythematosus. All were treated with carbamazepine. Improvement was evident within 2-14 days of initiation of low doses of carbamazepine (4-10 mg/kg daily). The plasma drug levels were 2.8-8.2 microg/mL (therapeutic antiepileptic range = 8-12 microg/mL). The chorea disappeared within 2-12 weeks. The duration of treatment was 1-15 months. No side effects were observed. Recurrence was observed in three patients who received a second trial of carbamazepine with a good response. We suggest that carbamazepine may serve as a first-line treatment for rheumatic chorea. PMID- 11020641 TI - Neuromotor spectrum of periventricular leukomalacia in children born at term. AB - The spectrum of neuromotor abnormalities of term children with periventricular leukomalacia (PVL) has never been specifically defined. We report 12 term children with PVL to delineate its long-term clinical correlates. A retrospective review of a standardized computer database and files of a single pediatric neurologist during a 7-year period was performed. The imaging studies were reviewed independently by two neuroradiologists. The mean age of the patients at the initial neurologic assessment was 24. 4 months (range 5-60); nine were males. The reason for the assessment was developmental delay in 10 (83.3%), seizure in one, and attention-deficit-hyperactivity disorder in one. Three children (25%) had normal motor examinations, three (25%) were hypotonic, three (25%) had spastic diplegia, two (16.7%) had spastic quadriplegia, and one (8.3%) had fine motor abnormalities. Nine children (75%) had developmental delay (severe global delay in six), and two children (16.7%) had epilepsy; electroencephalograms were abnormal in six (50%). PVL was mild in five (41.7%), moderate in two (16.6%), and severe in five (41.7%) children. Four of eight children with global developmental delay had enlargement of cerebral sulci in addition to white matter changes. PVL in term children presents with a spectrum of neurologic abnormalities, particularly developmental delay and heterogeneous motor findings not limited to classic spastic diplegia. The clinician should consider the diagnosis of PVL in the context of term children with developmental delay and motor abnormalities, even in the absence of perinatal difficulties. PMID- 11020642 TI - Lamotrigine monotherapy in children. AB - The effectiveness of lamotrigine as a monotherapeutic agent for a variety of pediatric epilepsies was reviewed retrospectively. Children were categorized as having focal vs generalized epilepsy and according to whether they were antiepileptic drug naive or drug exposed. Data collected included dosages, side effects, length of follow-up, number of prior drugs, and treatment response. Treatment was considered successful if the patient was seizure free for 6 months or more. Eighty-three children were identified (average age = 8.7 years); 43 had focal epilepsy, 32 had generalized epilepsy, and eight were not classified. Twenty-nine patients were classified as having specific syndromes. Fourteen patients were drug naive. The median follow-up period was 8 months (mean = 8.5). Overall, 45% were seizure free, 44% with focal epilepsy and 36% with generalized epilepsy. All children with juvenile myoclonic epilepsy and benign rolandic epilepsy of childhood were seizure free, although not all had been treated for at least 6 months. One third of drug-naive patients were seizure free. Rash was the most common side effect and was reported in five patients (6%); two patients discontinued the drug. None had Stevens-Johnson syndrome. One quarter of children experienced nonquantifiable improvements, namely increased alertness and improved behavior regardless of seizure control. Lamotrigine is effective as a monotherapeutic agent in children for both focal and generalized epilepsies. Side effects are relatively uncommon. Lamotrigine may be an effective firstline agent. PMID- 11020643 TI - Progressive facial hemiatrophy after epileptic seizures. AB - Intractable complex partial seizures developed in a 3-year-old female with normal intracranial findings on computed tomography. Frontal paramedian band-like depression of the skin gradually developed thereafter, and progressive facial hemiatrophy (Parry-Romberg syndrome) was diagnosed. Computed tomography scanning at 5 years of age revealed multiple parenchymal calcifications and low-density areas in the white matter of the frontoparietal lobes. Epileptic seizures, one of the major neurologic complications of progressive facial hemiatrophy, could precede the succeeding neurocutaneous changes. PMID- 11020644 TI - Left vagal nerve stimulation in six patients with hypothalamic hamartomas. AB - Six patients with medically refractory epilepsy secondary to hypothalamic hamartomas were treated with intermittent stimulation of the left vagal nerve. Three of the patients had remarkable improvements in seizure control. Four of these six patients had severe autistic behaviors. Striking improvements in these behaviors were observed in all four during treatment with intermittent stimulation. This finding suggests that vagal nerve stimulation can control seizures and autistic behaviors in patients with hypothalamic hamartomas. PMID- 11020645 TI - Abnormal myelination in a patient with deletion 14q11.2q13.1. AB - A male carrying an interstitial deletion of chromosome 14, presumably del(14)(q11.2q13), and presenting with abnormal myelination on magnetic resonance imaging is described. The abnormal myelination was evidenced as a high-signal intensity on T(2)-weighted magnetic resonance imaging. The patient had severe neurologic signs, various dysmorphic features, and a marked microcephaly. To our knowledge, this case is the first patient reported with abnormal myelination and a deletion of chromosome 14. PMID- 11020646 TI - Variable onset of metachromatic leukodystrophy in a Vietnamese family. AB - We report two siblings with metachromatic leukodystrophy, one who presented at 7 years of age (juvenile onset) and his sister who presented at 22 years of age (adult onset). They are compound heterozygotes for two novel mutations in the arylsufatase A gene (ARSA). The responsible mutations in this Vietnamese family consist of a missense mutation with 5% enzyme activity (R143G) and a nonsense mutation (W318ter), from which no enzyme activity would be expected. These mutations in the ARSA gene have not been previously reported and may be useful when diagnosing metachromatic leukodystrophy in other affected Vietnamese individuals. The variability in presentation suggests that the genotype alone is not sufficient to determine the onset and course of the disease and modifying genetic and perhaps nongenetic factors likely contribute. PMID- 11020647 TI - Acute disseminated encephalomyelitis associated with poliomyelitis vaccine. AB - A 6-year-old female patient with acute disseminated encephalomyelitis associated with poliomyelitis vaccine virus is reported. She had a history of high fever, headache, and gait disturbance. Neurologic examination confirmed spastic triparesis, urinary incontinence, diminution of tactile sensation, and vision deterioration. Hemography, serum laboratory findings, and urinalysis were normal. The cerebrospinal fluid was clear, with normal pressure, 9 leukocytes/mm(3), and 27 mg/dL protein, but the myelin basic protein was elevated to 10.7 ng/mL. T(2) weighted magnetic resonance imaging disclosed multifocal high-intensity lesions of the spinal cord. The serum polio virus type 2 antibody titer was raised in the acute phase, and polio vaccine virus type 2 was detected in viral cultures of the cerebrospinal fluid and pharynx swab and had undergone an A-G neurovirulence mutation at nucleotide 481. Finally, she had human leukocyte antigen (HLA)-Cw3 and HLA-DR2, to which multiple sclerosis is related in Japan. Thus the cause of ADEM may have been related to her HLA type. PMID- 11020648 TI - Anterior horn cell disease and olivopontocerebellar hypoplasia. AB - To date, fewer than 30 cases of anterior horn cell disease with associated olivopontocerebellar hypoplasia have been reported. We describe five patients and review the literature on this uncommon disorder. In addition to a syndrome of progressive spinal muscular atrophy similar to that seen in Werdnig-Hoffmann disease, this disorder is characterised by hypoplasia of the olivary nuclei, pons, and cerebellum. Additional clinical features may include dysmorphism, abnormal eye movements, stridor, congenital joint contractures, and enlarged kidneys. Pontocerebellar hypoplasia may be associated with posterior fossa cystic malformations, cerebral atrophy, and a demyelinating neuropathy. PMID- 11020649 TI - A female with central anticholinergic syndrome responsive to neostigmine. AB - Central anticholinergic syndrome is a rarely observed condition in children. The occurrence of this syndrome after ingestion of Solanum pseudocapsicum is infrequent because findings tend to be milder and localized to the gastrointestinal system, without central nervous system involvement. Most patients do not present with diagnostic problems because their relatives can usually report any ingestion of poisonous agents; however, when drug poisoning or plant ingestion is uncertain, a differential diagnosis with encephalitis must be considered. Physostigmine salicylate is the specific antidote because it crosses the blood-brain barrier because of its tertiary ammonium group. Neostigmine methylsulfate has a quaternary ammonium group, which prevents its penetration through the blood-brain barrier; hence its primary influence is believed to be due to its action on the peripheral nervous system. We describe a female with central anticholinergic syndrome caused by ingestion of Solanum pseudocapsicum. A slow intravenous infusion of neostigmine methylsulfate (0.03 mg/kg) immediately resolved the clinical picture. To our knowledge, this case is the first reported of central anticholinergic syndrome occurring after ingestion of Solanum pseudocapsicum in a child and the first report of a complete and rapid remission after intravenous neostigmine methylsulfate administration. PMID- 11020650 TI - Galactose metabolism and ovarian toxicity. AB - Galactose is an energy-providing nutrient and also a necessary basic substrate for the biosynthesis of many macromolecules in the body. Metabolic pathways for galactose are important not only for the provision of these pathways but also for the prevention of galactose and galactose metabolite accumulation. Problems with galactose metabolism can cause a variety of clinical manifestations in animals and humans. It has been found that the mammalian ovary is particularly susceptible to damage from the accumulation of galactose and galactose metabolites. The galactose metabolites Gal-1-P, galactitol, and UDPgal are all considered to be important in this toxicity and proposed mechanisms include interference with ovarian apoptosis and gonadotrophin signaling. This review addresses the most recent scientific findings regarding the possible mechanisms of galactose-induced ovarian toxicity and also the possible protective role of hormonal and antioxidant therapy. In addition, the available epidemiologic and scientific evidence linking galactose intake with risk of ovarian cancer is discussed. PMID- 11020651 TI - Rat two-generation reproduction and dominant lethal study of acrylamide in drinking water. AB - Fischer 344 (F344) F(0) weanling rats, 30/sex/group, were exposed to acrylamide in drinking water at 0.0, 0.5, 2.0, or 5.0 mg/kg/day for 10 weeks and then mated. Exposure of F(0) females continued through gestation and lactation of F(1) litters. F(0) males, after F(0) mating, were removed from exposure and mated (one male: two untreated females) for the dominant lethal (DL) assay. Thirty F(l) weanlings/sex/group were exposed for 11 weeks to the same dose levels as their parents, and then mated to produce F(2) offspring. F(0) and F(l) parents and F(1) and F(2) weanlings were necropsied. Prebreeding exposure of F(0) and F(l) animals resulted in systemic toxicity at 2.0 to 5.0 mg/kg/day, with head tilt and/or foot splay increased at 0.5 to 5.0 mg/kg/day. F(0) and F(l) reproductive indices and gestational length were unaffected. Implantations and live pups/litter at birth were reduced at 5.0 mg/kg/day. Survival of F(l) and F(2) pups was reduced at 5.0 mg/kg/day for PND 0 through 4 only. In the DL assay, total and live implants were reduced, pre- and postimplantation loss was increased, and the frequency of DL factors (F(L)%) was increased at 5.0 mg/kg/day. At 5.0 mg/kg/day, adult F(l) male peripheral nerves exhibited axonal fragmentation and/or swelling; F(l) female spinal cord sections were unremarkable. The NOEL for prenatal DL was 2.0 mg/kg/day; the NOEL for adult systemic toxicity, including neurotoxicity, was < or = 0.5 mg/kg/day. Therefore, neurotoxicity and DL were differentially affected. PMID- 11020652 TI - Effects of prenatal and postnatal exposure to styrene dimers and trimers on reproductive function in rats. AB - Styrene dimers and trimers (SDT) were evaluated for reproductive toxicity in Sprague-Dawley rats. SDT was administered orally to rats at doses of 0, 0.04, 0.2, and 1.0 mg/kg from day 6 of gestation through day 21 after delivery. Clinical signs, including pregnancy and lactation, and changes in body weight and food consumption were assessed. All dams underwent a gross necropsy examination. The brain, liver, kidney, ovary, uterus, thyroid gland, and pituitary were weighed. Offspring were evaluated for the effects of the test compound on viability, growth, anogenital distance, preputial separation and vaginal opening, behavioral function, estrous cycling, mating, and fertility. There were no test compound-related clinical signs or effects on body weight or food consumption in dams during any phase of the study. In addition, no abnormalities in delivery or lactation, including gestation length, were noted in any dam. No dose-dependent changes were observed in pup viability or growth. There were no adverse effects of SDT on any developmental landmark, learning, memory, or estrous cycling in offspring. The number of days to inseminations in the 0.2 mg/kg group was significantly greater than that in the control group, but was independent of dose. No test compound-related necropsy findings were seen in either the dams or the offspring. No compound-related histopathologic findings were noted in the reproductive tissues of either the male or female offspring. No compound-related alterations in sperm motion or density were detected in the offspring. Thyroid stimulating hormone levels in the male offspring of the 0.2 mg/kg and 1.0 mg/kg groups were significantly higher than those in the controls, whereas thyroid hormone (T(3), T(4)) levels in these groups were comparable to the controls. In addition, the thyroid glands of males in all groups were similar histologically. These results indicate that SDT administered at doses as high as 1.0 mg/kg (1000 times the estimated human daily intake) did not produce reproductive toxicity in dams or offspring. PMID- 11020653 TI - Developmental toxicity of butyl benzyl phthalate in the rat using a multiple dose study design. AB - The developmental toxicity of butyl benzyl phthalate (BBP) was investigated in the rat using ten dose groups between 270 and 2100 mg/kg/day. Exposure was by daily gavage from gestation day 5 through 16 or gestation day 5 through 20. Dose response data were analyzed using the benchmark approach by fitting dose-response models to the various endpoints. BBP induced increased liver and kidney weights in dams, accompanied by liver enzyme increases in maternal serum. Extramedullary hematopoiesis, which was already substantial in control pregnant animals, was increased after BBP treatment. Fetotoxicity included increased resorptions, reduced fetal weights, increased incidence of skeletal anomalies, and reduced fetal testis weights in the presence of an increased incidence of retarded testicular descent. As embryotoxicity was found at lower dosages compared to observed maternal toxicity, BBP appeared to be a specifically embryotoxic compound. The extended exposure protocol (gestation day 5 through 20) appeared more sensitive for measuring fetotoxic effects. We recommend the use of more doses in toxicity tests, together with the benchmark approach as an appropriate and more accurate method for analyzing dose-response data compared to the NOAEL approach. PMID- 11020654 TI - Two-generation reproduction toxicity studies of di-(C(7)-C(9) alkyl) phthalate and di-(C(9)-C(11) alkyl) phthalate in the rat. AB - Di-(C(7)-C(9) alkyl) phthalate (D79P) and di-(C(9)-C(11) alkyl) phthalate (D911P), based on high-normality linear oxo-alcohols, have been assessed for their impact upon reproductive performance in Sprague-Dawley rats. Rats were continuously exposed to either D79P or D911P at dietary levels of 0%, 0.1%, 0.5%, or 1.0% over two generations. Selected F(0) offspring (F(1) generation) were exposed to the same dietary concentration of D79P or D911P as the respective F(0) animals, and were mated to produce F(1) offspring. Both D79P and D911P markedly reduced body weight gain in F(0) and F(1) adult males at the highest dose, but females were affected to a lesser extent. There was no impairment of fertility, fecundity, or development in either generation, but body weights of offspring in the 1.0% D79P and 1.0% D911P groups were slightly and transiently reduced over the weaning period. Although decreases in the weight of several organs were accounted for by depressed body weight, ovary weights were reduced in both generations exposed to 1.0% D79P, and epididymidal weights were slightly reduced in adults of both generations exposed to 1.0% D911P. However, ovarian function assessed by the oestrus cycle and mating behaviour-and epididymidal sperm concentration, motility, and morphology were unaffected by either substance. Treatment resulted in liver changes, particularly in males, characterised by increased liver weight in young animals, histopathologic changes and reduced organ weight in mature animals, and an increase in palmitoyl CoA oxidase activity. In conclusion, neither D79P nor D911P impaired reproductive function in rats when administered in the diet at levels that induce systemic toxicity, and the NOAEL for effects on reproduction in the rat is 0.5% for both D79P and D911P. PMID- 11020655 TI - Adverse effects of dibutyltin dichloride on initiation and maintenance of rat pregnancy. AB - The present study was conducted to evaluate the adverse effects of dibutyltin dichloride (DBTCl) on initiation and maintenance of pregnancy after maternal exposure during early pregnancy in rats. After successful mating, female rats were given DBTCl by gastric intubation on Days 0 to 3 or on Days 4 to 7 of pregnancy at 0, 3.8, 7.6, or 15.2 mg/kg. Food-restricted pregnant rats were given an amount of feed equal to the feed intake of female rats treated with DBTCl at 15.2 mg/kg on Days 0 to 3 or on Days 4 to 7 of pregnancy. Female rats were sacrificed on Day 20 of pregnancy and pregnancy outcome was determined. After administration of DBTCl on Days 0 to 3, the rate of nonpregnant females and the incidence of preimplantation embryonic loss in the 7.6 mg/kg group were significantly higher than those in the control group, and those in the 15.2 mg/kg group were significantly higher than those in the control and pair-fed groups. In females with implantations, the numbers of implantations and live fetuses and the incidence of postimplantation embryonic loss in the groups given DBTCl on Days 0 to 3 were not significantly different from those in the control group. The incidence of postimplantation embryonic loss in the groups given DBTCl on Days 4 to 7 at 7.6 and 15.2 mg/kg was significantly higher than that in the control and pair-fed groups. It can be concluded that DBTCl adversely affects initiation and maintenance of pregnancy when administered during early pregnancy and that the manifestations of the adverse effects of DBTCl vary with the gestational stage at the time of maternal exposure. PMID- 11020658 TI - Forum on oxidative stress status (OSS) and its measurement PMID- 11020656 TI - Reproductive toxicology of acephate in male mice. AB - The reproductive toxicity of the insecticide acephate was studied in male mice. Adult male mice were treated by gavage with acephate at doses of 0, 7, 14, and 28 mg/kg/day for 4 weeks before mating with untreated females. Signs of cholinergic effects were observed in the 28 mg/kg/day group. Brain and skeletal muscle acetylcholinesterase activity was inhibited only in this group. Acephate treatment was associated with a decreased number of implantations and live fetuses, and an increased number of early resorptions at 28 mg/kg/day. The percent morphologically normal spermatozoa was unaffected in all dose groups; however, sperm motility and count were decreased in the 14 and 28 mg/kg/day groups compared to the control. Histologic examination of brain did not reveal any abnormalities. Dose related histologic changes, including degeneration of muscle fibers, were observed in the muscles of male mice treated with any of the doses of acephate. The current study demonstrated adverse effects of male acephate exposure on pregnancy outcome with effects on sperm parameters at 14 and 28 mg/kg/day. PMID- 11020657 TI - Folic acid and human malformations: a summary and evaluation. AB - A large body of evidence gathered over the past 30 or more years has led to the firmly established belief that deficiency of the vitamin folic acid is a cause of congenital malformations of neural tube closure. Beginning with studies showing folic acid levels of mothers of children with such defects to be low, this belief has been solidified by epidemiologic studies revealing that this consequence is prevented by maternal supplements of the vitamin from early pregnancy. The present article reviews this evidence for the purpose of examining the claims of the efficacy of folic acid in this respect. This seems to be an advantageous moment to do so, because no clear impact of folic acid supplementation and fortification on the prevalence of neural tube defects has as yet been documented; and furthermore a pause seems to have been reached in such studies. It is felt that a historical, i.e. a chronologic approach will best describe the findings, and therefore they will be considered as they unfolded. PMID- 11020659 TI - Detection of superoxide anion released extracellularly by endothelial cells using cytochrome c reduction, ESR, fluorescence and lucigenin-enhanced chemiluminescence techniques. AB - Endothelium produces oxygen-derived free radicals (nitric oxide, NO&z.rad;; superoxide anion, O(2)(*-)) which play a major role in physiology and pathology of the vessel wall. However, little is known about endothelium-derived O(2)(*-) production, particularly due to the difficulty in assessing O(2)(*-) when its production is low and to controversies recently raised about the use of lucigenin enhanced chemiluminescence. We compared four techniques of O(2)(*-) assessment when its production is low. In the present study, we have compared ferricytochrome c reduction, electron spin resonance (ESR) spectroscopy using DMPO as spin trap, hydroethidine fluorescence, and lucigenin-enhanced chemiluminescence to assess O(2)(*-) production in cultured bovine aortic endothelial cells (BAEC). We focused our study on extracellular O(2)(*-) production because the specificity of the signal is provided by the use of superoxide dismutase, and this control cannot be obtained intracellularly. We found that the calcium ionophore A23187 dose-dependently stimulated O(2)(*-) production, with a good correlation between all four techniques. The signals evoked by postconfluent BAEC were increased 2- to 7-fold in comparison to just confluent BAEC, according to the technique used. Ferricytochrome c 20 microm rather than at 100 microm appears more suitable to detect O(2)(*-). However, in the presence of electron donors such as NADH or NADPH, lucigenin-enhanced chemiluminescence generated high amounts of O(2)(*-). Thus, ferricytochrome c reduction, electron spin resonance (ESR), and hydroethidine fluorescence appear as adequate tools for the detection of extracellular endothelium-derived O(2)(*-) production, whereas lucigenin may be artifactual, even when a low concentration of lucigenin is employed. PMID- 11020660 TI - Kinetic analysis of lipid-hydroperoxides in plasma. AB - We increased the precision of chemiluminescent procedure for measuring lipid hydroperoxides in plasma or lipoproteins by (i) escaping from extraction and chromatography of lipids, (ii) using detergent dispersed lipids, and (iii) calculating the results by fitting the photon emission rate with the integrated equation, which describes the model of the series of reactions. The use of kinetics instead of the crude integration of cps increases precision because at each measurement the correct reaction pathway is tested. This was relevant for the optimization of the analytical procedure, contributing to the elimination of possible side reactions. The relationship between lipid hydroperoxide content in the sample and cps is not linear; thus, the calculation of results through internal calibration is carried out using an exponential equation. This is in agreement with the reaction mechanism and raises the point of the linear calibration previously reported in other chemiluminescent procedures. Although sensitive and precise, this procedure suffers for being time consuming, requiring approximately 30 min per sample. Moreover, since no chromatography is used, information about the hydroperoxides in different lipid classes is missing. Obviously this will be solved when a validated procedure for quantitatively extracting lipid hydroperoxides is available. PMID- 11020661 TI - Biomarkers of myeloperoxidase-derived hypochlorous acid. AB - Hypochlorous acid is the major strong oxidant generated by neutrophils. The heme enzyme myeloperoxidase catalyzes the production of hypochlorous acid from hydrogen peroxide and chloride. Although myeloperoxidase has been implicated in the tissue damage that occurs in numerous diseases that involve inflammatory cells, it has proven difficult to categorically demonstrate that it plays a crucial role in any pathology. This situation should soon be rectified with the advent of sensitive biomarkers for hypochlorous acid. In this review, we outline the advantages and limitations of chlorinated tyrosines, chlorohydrins, 5 chlorocytosine, protein carbonyls, antibodies that recognize HOCl-treated proteins, and glutathione sulfonamide as potential biomarkers of hypochlorous acid. Levels of 3-chlorotyrosine and 3,5-dichlorotyrosine are increased in proteins after exposure to low concentrations of hypochlorous acid and we conclude that their analysis by gas chromatography and mass spectrometry is currently the best method available for probing the involvement of oxidation by myeloperoxidase in the pathology of particular diseases. The appropriate use of other biomarkers should provide complementary information.Keywords-Free radicals, Myeloperoxidase, Neutrophil oxidant, Hypochlorous acid, Chlorotyrosine, Chlorohydrin, Oxidant biomarker PMID- 11020662 TI - Sensitive and nonenzymatic measurement of hydrogen peroxide in biological systems. AB - The increasing demand in detecting H(2)O(2) under various experimental conditions is only partly fulfilled by most conventional peroxidase-based assays. This article describes a sensitive and nonenzymatic H(2)O(2) assay that is based on the chemiluminescence reaction of luminol with hypochlorite. It allows the determination of H(2)O(2) down to nanomolar concentrations. Actual H(2)O(2) concentrations rather than a turnover of H(2)O(2) can be determined in monolayer cultures, perfusates, suspensions of intact cells, organelles, and crude homogenates. One of the strengths of this assay is that it may be used to assess fast enzyme kinetics (catalase, glutathione peroxidase, oxidases) at very low H(2)O(2) concentrations. Its use together with a glucose oxidase/catalase system appears to be a powerful tool in studying signal functions of H(2)O(2) in various biological systems on a quantitative basis. Several applications are discussed in detail to demonstrate the technical requirements and analytical potentials. PMID- 11020663 TI - NADPH oxidase: a universal oxygen sensor? AB - NADPH oxidase is classically regarded as a key enzyme of neutrophils, where it is involved in the pathogenic production of reactive oxygen species. However, NADPH oxidase-like enzymes have recently been identified in non-neutrophil cells, supporting a separate role for NADPH-oxidase derived oxygen species in oxygen sensitive processes. This article reviews the current literature surrounding the potential role of NADPH oxidase in the oxygen sensing processes which underlie hypoxic pulmonary vasoconstriction, systemic vascular smooth muscle proliferation, carotid and airways chemoreceptor activation, erythropoietin gene expression, and oxytropic responses of plant cells. PMID- 11020664 TI - Relative reactivity of lysine and other peptide-bound amino acids to oxidation by hypochlorite. AB - Antibacterial and inflammatory responses of neutrophils and macrophages produce hypochlorite as a major oxidant. Numerous side chains of amino acids found in extracellular proteins can be modified by hypochlorite, including His, Arg, Tyr, Lys, Trp, and Met. We studied the relative reactivity of each of these amino acid residues in short N-blocked peptides, where other residues in the peptide were highly resistant to hypochlorite attack. Hypochlorite treatment led to modified peptides in each case, which were detected by changes in retention on reversed phase HPLC. A distinct single product, consuming two equivalents of hypochlorite per equivalent of peptide, was obtained from the Lys-containing peptides. UV spectroscopy, nuclear magnetic resonance (NMR), and electrospray/mass spectroscopy identified this product as the dichloramine at the epsilon-amino group of the Lys side chain. The dichloramine at Lys did not decompose to form a detectable amount of carbonyl reactive with dinitrophenylhydrazine. The dichloramine at Lys did however quantitatively revert back to Lys during HCl digestion of the tetrapeptide for amino acid analysis, with simultaneous modification of the adjacent Phe residue. The formation of the dichloramine at Lys was not blocked by peptides or acetylated amino acids that contained Tyr, His, or Arg. In contrast, the presence of equimolar Met-containing peptide, or N Acetyl-Trp, both inhibited the formation of the dichloramine at Lys. Thus, Met and Trp side chains of proteins might be able to protect Lys from chloramine formation under some circumstances, but this interpretation must consider that Met and Trp are typically found in relatively inaccessible hydrophobic sites, whereas lysine is typically exposed on the protein surface. The hierarchy of amino acid reactivities examined here will aid in the prediction of residues in biological samples most likely to be modified by hypochlorite. PMID- 11020665 TI - How should xanthine oxidase-generated superoxide yields be measured? AB - The rate of formation of superoxide measured by its reduction of tetranitromethane (TNM) and by its reduction of ferric cytochrome c (Fe(III) cc) are in excellent agreement when the superoxide is generated from a simple chemical precursor. In contrast, the rate of formation of superoxide generated in the reaction of xanthine oxidase with acetaldehyde is much higher (up to a factor of 6) when measured with TNM and compared with Fe(III) cc. It is shown that Fe(III) cc measures superoxide that has diffused from the enzyme, and that TNM probably scavenges all the dioxygen that is reduced by one electron by the enzyme. The TNM traps enzyme-bound superoxide in competition with the second electron transfer and proton transfer, which normally yield hydrogen peroxide. The proton transfer is probably rate determining, k(p) 80 years of age, respectively. Only 22% of the normal aged controls carried the 4977 bp mtDNA deletion, whereas 77% (17/22) of the ESRD patients exhibited the mtDNA deletion. Using a semiquantitative PCR method, we determined the proportion of the 4977 bp-deleted mtDNA from the muscles that had been confirmed to harbor the deletion. We found that the proportions of the 4977 bp-deleted mtDNA in the muscle were significantly higher than those of the aged matched controls. Using long-range PCR techniques, a distinctive array of mtDNA deletions was demonstrated in the muscle of uremic patients. In summary, we found diverse and multiple mtDNA deletions in the skeletal muscles of ESRD patients. These deletions are more prevalent and abundant in ESRD patients than those found in normal populations. Accumulation of uremic toxins and impaired free radical scavenging systems may be responsible for the increased oxidative stress in ESRD patients. Such stress may result in oxidative damage and aging-associated mutation of the mitochondrial genome. PMID- 11020668 TI - Dediazoniation of p-hydroxybenzenediazonium ion in a neutral aqueous medium. AB - The dediazoniation of p-hydroxybenzenediazonium ion (PDQ) in a neutral aqueous medium has been studied under controlled experimental conditions to prevent photochemical and/or heterolytic side-reactions. Oxygen increased the dediazoniation rate of PDQ and caused the appearance of quinone and hydroquinone. An accumulation of quinone deriving from the reduction of PDQ by hydroquinone was also observed. In ESR analyses with different spin traps, the most stable signal was identified as belonging to the adduct of the p-hydroxyphenyl radical, even in the presence of dimethylsulfoxide or ethanol. A general scheme is proposed including three pathways for the homolytic dediazoniation of PDQ. Pathway 1 represents dediazoniation induced by a hydroxyl ion, a slow process at neutral pH and an even slower one with deaerated samples; a favored quinoid structure is put forward to explain these results. In pathway 2, the formation of a semiquinone radical via the reaction of an aryl radical with oxygen is put forward to justify the increase in the dediazoniation rate in the presence of oxygen. In pathway 3, hydroquinone, produced by semiquinone dismutation, may act as a reducing agent. PMID- 11020670 TI - Erratum(a) PMID- 11020669 TI - Hemiketal formation of dehydroascorbic acid drives ascorbyl radical anion disproportionation. AB - In this paper the relationship between the ascorbate anion (AH(-)) and its oxidation products, ascorbyl radical anion (A&z.rad;(-)) and dehydroascorbic acid (DHA), are studied by means of theoretical calculations. Additional calculations are performed on alpha-hydroxytetronate, a model compound of ascorbate lacking the side chain. The method uses density functional theory with the B3LYP functional and a polarizable conductor dielectric model to compute solvation effects. Our results indicate that the model compound reacts with the alpha tocopheroxyl radical to regenerate vitamin E with a free energy change of reaction (in water) of -7.4 kcal/mol. This reaction is 2.9 kcal/mol more exergonic than the corresponding reaction involving ascorbate, suggesting that the model compound may make a more effective antioxidant than ascorbate. However, the disproportionation of the ascorbyl radical anion, a reaction that regenerates AH(-), is found to be exergonic while the similar reaction involving the model compound is slightly endergonic. The reason for the difference is that the disproportionation of A&z. rad;(-) is found to be driven by the formation of the hemiketal structure of dehydroascorbic acid (DHA). PMID- 11020671 TI - Berstein's anti-reductionistic materialism: On the road towards a biology of activity (1965). AB - Bernstein's paper, "On the Road Towards a Biology of Activity," appeared the year before his death.2 With this paper, Bernstein closed several lines of argument that he had been developing from the onset of his career in the early 1920s. The paper converges on the notion of activity. In accordance with his own shifting focus heuristic (cf. Bongaardt, 1996), Bernstein challenged future researchers of movement to integrate models of the movement functions that constitute activity. He suggested that these functions are: the coordination of movement, the planning of movement, and the exploration of better, optimal ways to move. In the 1920s, Bernstein had collaborated with his friend and colleague L.S. Vygotsky at the Moscow Institute of Experimental Psychology. Vygotsky (cf. 1926/1994) was the first to place activity at the core of Soviet psychology. According to Vygotsky, reflexology and behaviorism, then dominant approaches in psychology, were fundamentally wrong; they focus on building-blocks of behavior without addressing phenomena that stand out as typically human, most importantly, consciousness. Rather than starting with building blocks, psychology should start with the daily activity of human beings in their environment and show how this activity relates to consciousness. Forty years later, in his 1965 paper, Bernstein stressed a point that mirrors Vygotsky's: Reflexes are not building blocks of movement. The general characteristics of any movement precede the specificity of such units, whether reflexes or synergies, and this primacy pertains to the actual organization of movements as well as to the study of movement. The development and relevance of the activity concept in Bernstein's work in the period from 1925 to 1965 deserves a study of its own; here, a brief historical sketch of Bernstein's activity concept is offered, along with a few theoretical considerations concerning activity's constituent functions. PMID- 11020672 TI - Vibration-induced postural reaction continues after the contact with additional back support. AB - We investigated the development of postural reactions induced in standing subjects by Achilles tendon vibration. We compared vibratory reactions in 3 different conditions: normal standing, standing near support, and when the solid support being protracted forward changed the initial posture. Additional support for the back was placed at subject's sacral or shoulder level. In the easy standing condition, the postural vibration reaction consists of progressive backward upper body movement. When the body contacted the additional support on the sacral level during the vibratory reaction, the movement of the upper body continued in most of the subjects. This was accompanied by an increase of pressure on the toes. When the support was applied at the shoulder level, the body motion reversed its direction in half of the subjects. In this case, backward-forward oscillations occurred near the support. The initial change of body-support interaction did not influence the ensuing vibration reaction; namely the reaction was similar to that with the support near to the body at the sacral level. Our data demonstrate that the vibration-induced reaction is not a local reaction limited to one joint, but a complex postural synergy that involves both leg and trunk muscles and integrates the information from touch and pressure afferents of the upper body. PMID- 11020673 TI - Role of different sensory inputs for maintenance of body posture in sitting rat and rabbit. AB - In this paper, we describe the postural activity in sitting rats and rabbits. An animal was positioned on the platform that could be tilted in the frontal plane for up to +/-20-30 degrees, and postural corrections were video recorded. We found that in both rat and rabbit, the postural reactions led to stabilization of the dorsal-side-up trunk orientation. The result of this was that the trunk tilt constituted only approximately 50% (rat) and 25% (rabbit) of the platform tilt. In addition, in the rabbit the head orientation was also stabilized. Trunk stabilization persisted in the animals subjected to the bilateral labyrinthectomy and blindfolding, suggesting that the somatosensory input is primarily responsible for trunk stabilization. Trunk stabilization was due to extension of the limbs on the side moving down, and flexion of the opposite limbs. EMG recordings showed that the limb extension was caused by the active contraction of extensor muscles. We argue that signals from the Golgi tendon organs of the extensor muscles may considerably contribute to elicitation of postural corrective responses to the lateral tilt. PMID- 11020674 TI - Against a final ballistic process in the control of voluntary action: evidence using the Hoffmann reflex. AB - Reduced surface electromyogram (EMG) onsets, observed in some cases when trying to stop an earlier intended action before it begins, were taken to suggest a control process that remains open to stopping right up to the point of motor discharge. This interpretation argues against a final ballistic (involuntary) process in the control of voluntary action. That a final ballistic process may receive reduced input shortly before its manifestation as reduced output (i.e., motor discharge), however, provides for an alternative interpretation of these same data. In this study we used the Hoffmann (H) reflex to further investigate for stopping effects in the brief interval before voluntary motor discharge. Late stopping effects on the facilitated H-reflex within the time window that a final ballistic process would otherwise be expected (i.e, shortly before EMG onset) were observed in some instances. We conclude from these data good evidence against a final ballistic process in the control of voluntary action. PMID- 11020675 TI - Effects of cervical spine immobilization technique and laryngoscope blade selection on an unstable cervical spine in a cadaver model of intubation. AB - STUDY OBJECTIVE: Orotracheal intubation (OTI) is commonly used to establish a definitive airway in major trauma victims, with several different cervical spine immobilization techniques and laryngoscope blade types used. This experimental, randomized, crossover trial evaluated the effects of manual in-line stabilization and cervical collar immobilization and 3 different laryngoscope blades on cervical spine movement during OTI in a cadaver model of cervical spine injury. METHODS: A complete C5-C6 transection was performed by using an osteotome on 14 fresh-frozen cadavers. OTI was performed in a randomized crossover fashion by using both immobilization techniques and each of 3 laryngoscope blades: the Miller straight blade, the Macintosh curved blade, and the Corazelli-London-McCoy hinged blade. Intubations were recorded in real time on fluoroscopy and then transferred to video and color still images. Outcome measures included movement across C5-C6 with regard to angulation expressed in degrees of rotation and axial distraction and anteroposterior displacement with values expressed as a proportion of C5 body width. Cormack-Lehane visualization grades were also recorded as a secondary outcome measure. Data were analyzed by using multivariate analysis of variance to test for differences between immobilization techniques and between laryngoscope blades and to detect for interactions. Significance was assumed for P values of less than.05. RESULTS: Manual in-line stabilization resulted in significantly less movement than cervical collar immobilization during OTI with regard to anteroposterior displacement. Use of the Miller straight blade resulted in significantly less movement than each of the other 2 blades with regard to axial distraction. The Cormack-Lehane grade was significantly better with manual in-line stabilization versus cervical collar immobilization; no differences were observed between blades. CONCLUSION: Manual in-line stabilization results in less cervical subluxation and allows better vocal cord visualization during OTI in a cadaver model of cervical spine injury. The Miller laryngoscope blade allowed less axial distraction than the Macintosh or Corzelli-London-McCoy blades. The clinical significance of this degree of movement is unclear. PMID- 11020676 TI - A randomized controlled trial on the effect of educational interventions in promoting airway management skill maintenance. AB - STUDY OBJECTIVE: This study was conducted to determine the natural history of airway management skill decay and examine the effect of independent practice and periodic feedback on airway management skill maintenance. METHODS: This prospective, randomized controlled study conducted at Dalhousie University in Halifax, Nova Scotia, Canada, between November 1997 and September 1998. A convenience sample of 84 health sciences students with no prior airway management experience was used. Participants were trained using an advanced airway manikin and then were randomly assigned to control (n=24), periodic feedback only (n=30), and independent practice plus periodic feedback (n=30) groups. Performance was measured by a 52-point weighted checklist at 0, 16, 25, and 40 weeks after the initial program. RESULTS: Group scores were analyzed using a mixed-model repeated measures analysis of variance and Bonferroni-adjusted P values. Overall group (P =.0002) and time (P =.0001) effects were significant. At time 0, there was no statistical difference in mean scores between groups (range 45.0 to 45.2). Control group performance fell over the first time interval (0 to 16 weeks) (mean score=34.0, P =.002) and remained lower at all intervals without further significant change. Scores in the independent practice plus feedback group revealed no significant changes over time and were significantly higher than the control group throughout. Performance in the periodic feedback only group showed a nonsignificant trend to improved performance over the control group. CONCLUSION: Airway management skill performance declines early after initial training. Independent practice combined with periodic feedback was effective in maintaining performance scores in an advanced airway management simulation. Periodic evaluation with feedback alone showed a nonsignificant trend toward improvement over control. PMID- 11020677 TI - Intravenous Lorazepam versus dimenhydrinate for treatment of vertigo in the emergency department: a randomized clinical trial. AB - STUDY OBJECTIVE: To determine whether lorazepam is more effective than dimenhydrinate in relieving the symptom of vertigo in the emergency department setting. METHODS: A prospective, randomized, double-blind trial of intravenous lorazepam versus dimenhydrinate therapy was conducted in the ED of a county owned, university-affiliated hospital. All adult patients who presented between January 24, 1998, and May 23, 1999, with the symptom of vertigo were eligible for inclusion. The intervention was varying the intravenous treatment between lorazepam, 2 mg, and dimenhydrinate, 50 mg. All patients received intravenous infusion of Ringer's lactate solution at a rate of 100 mL/h. Adequacy of randomization to the 2 treatment groups was assessed by comparing the patients' relevant baseline history, physical examination, and symptoms. The predetermined primary outcome measurement was the patient's sensation of "vertigo with ambulation" 1 and 2 hours after treatment. Secondary outcome measurements included vertigo while lying, sitting, and turning the head, ability to ambulate as judged by the enrolling physician, and sensation of nausea and drowsiness 1 and 2 hours after treatment, and whether the patient was "ready to go home" per patient report or physician assessment 2 hours after treatment. All patient symptoms were reported on 10-point scales. Outcome measurements were compared between the 2 treatment groups with a 2-way repeated-measures analysis of variance, Student's t test, Mann-Whitney U, and chi(2) test as appropriate. RESULTS: Ten patients refused entry into the study, 16 were excluded, and 74 were enrolled, treated, and included in the analysis. One enrolled patient had evidence of vertigo of central origin. The pretreatment values of vertigo with ambulation were strongly correlated with the patient's ability to ambulate (P <.001), suggesting good internal validity. The patients randomly assigned to the lorazepam group were sicker based on their pretreatment symptoms and ability to ambulate, and this may have biased the study results. The patients' symptom of "vertigo with ambulation" decreased 1.5 units more (95% confidence interval [CI] 0 to 3.0) on average on a 10-point scale 2 hours after treatment in the dimenhydrinate group. All other measures of vertigo also decreased more in the dimenhydrinate group, although the differences were not statistically significant. At 2 hours after treatment, the patients' ability to ambulate was better in the dimenhydrinate group (P <.001), and 17% (95% CI -2 to 36) more patients in this group were "ready to go home." Patients in the lorazepam group experienced a 1.8-unit (95% CI 0.2 to 3.4) greater increase in drowsiness 2 hours after treatment. CONCLUSION: Our results suggest that dimenhydrinate was more effective in relieving vertigo and less sedating than lorazepam at the intravenous doses administered in this study. Dimenhydrinate appears to be the preferred medicine for patients who present to the ED with vertigo likely to be of peripheral origin. PMID- 11020678 TI - Antibiotic prescribing for patients with colds, upper respiratory tract infections, and bronchitis: A national study of hospital-based emergency departments. AB - STUDY OBJECTIVE: Antibiotics are often used to treat viral upper respiratory tract infections, even though they are usually ineffective. However, frequent inappropriate antibiotic use contributes to the emergence of drug-resistant bacterial pathogens. This study used a national database to evaluate antibiotic use in treating upper respiratory tract infections in emergency departments. METHODS: Data were obtained from the 1996 National Hospital Ambulatory Medical Care Survey. Antibiotic prescribing rates were examined for colds, upper respiratory tract infections, and acute bronchitis. Patients with comorbid conditions or secondary diagnoses, such as chronic obstructive pulmonary disease, pneumonia, sinusitis, and HIV, were excluded. Bivariate and multivariate analyses were used to assess predictors of antibiotic use. RESULTS: Overall, there were an estimated 2.7 million ED visits for colds, upper respiratory tract infections, and bronchitis by children and adults in 1996. Antibiotics were prescribed for 24.2% (95% CI 18.9, 29.5) of patients with common colds and upper respiratory tract infections and for 42.2% (95% CI 35.2, 49.2) of patients with bronchitis. There were no significant associations between antibiotic use and patient race, sex, Hispanic ethnicity, geographic location, or source of payment. Antibiotics were prescribed less often by interns or residents than by staff or other physicians (odds ratio 0.43; 95% CI 0.19, 0.98), and patients younger than 18 years were less likely to receive antibiotics than adults (odds ratio 0.32; 95% CI 0.20, 0.52). Smokers were 4.3 (95% CI 2.2, 8.3) times more likely to receive antibiotics than nonsmokers. CONCLUSION: Antibiotics are commonly prescribed for ED patients with upper respiratory tract infections even though they are usually ineffective in otherwise healthy adults. Efforts should be made to reduce inappropriate antibiotic use for the sake of containing costs, preventing side effects, and limiting the spread of antibiotic resistance. PMID- 11020679 TI - Paramedic success rate for blind nasotracheal intubation is improved with the use of an endotracheal tube with directional tip control. AB - STUDY OBJECTIVES: Blind nasotracheal intubation (BNTI) is used to secure the airway in patients who are spontaneously breathing. The success rate for BNTI is often lower than for orotracheal intubation. We conducted this study to determine whether the use of an endotracheal tube (ETT) capable of directional tip control can improve the BNTI success rate. METHODS: This prospective, experimental study was conducted by a state emergency medical services agency during 1997, 1998, and 1999. Consecutive patients undergoing attempted BNTI or orotracheal intubation were included. Five paramedic units were trained to use an ETT with triggeractivated distal tip directional control for BNTIs (intervention group). Ten units used conventional ETTs for BNTIs and served as concurrent controls (control group). Subjects in the 2 groups were enrolled concurrently with nonrandomized allocation based on the agency providing service. An intubation attempt was defined by tube passage, and success was defined as confirmed endotracheal placement. RESULTS: A total of 219 BNTIs were studied (141 in the control group and 78 in the intervention group). BNTI was successful in 82 (58%) of 141 cases using conventional ETTs, and in 56 (72%) of 78 cases using directional tip control (P =.04). The overall success rate was 63%. CONCLUSION: Use of ETTs with distal directional control is associated with a higher success rate for BNTI than conventional ETTs. Use of ETTs with directional tip control significantly improves the success rates for BNTIs. PMID- 11020680 TI - Injury control in Honduras: a survey of injury mortality. AB - STUDY OBJECTIVE: We sought to evaluate the methods and accuracy of mortality data collection and summarize the injury mortality rate in one sector of the State of Olancho, Honduras, with the intent to establish a baseline of injury mortality that will identify potential areas of intervention and serve as a comparison after subsequent interventions. METHODS: Mortality data were collected from a rural, regional, health center database containing age, sex, and cause of death for one geographic sector in the State of Olancho, Honduras. Causes of death were classified as medical or intentional versus nonintentional injury. RESULTS: Accurate mortality data were difficult to obtain for several reasons: (1) deaths are often recorded by untrained health care workers, (2) causes of death are not coded in a standard manner, and (3) infant mortality is underreported. We found 132 recorded noninfant deaths. A disproportionate number of these resulted from injury, especially from intentional injury, particularly among male subjects aged 12 to 49 years. Eighty-two percent of male subjects aged 12 to 49 years who died did so from injuries, and 52% died from intentional injuries. Overall, 48% of all male deaths were injury related. The estimated male mortality rate (age 12 to 49 years) from injuries was 4.5 times that of the United States. CONCLUSION: Injury, particularly intentional injury, is an important cause of mortality in rural Honduras, particularly among male subjects aged 12 to 49 years. This suggests a fertile opportunity for intervention. More reliable data collection will be necessary to accurately target which specific causes of injury death are most amenable to interventions and to monitor the effect of injury control programs. PMID- 11020681 TI - Adults and children in severe motor vehicle crashes: a matched-pairs study. AB - STUDY OBJECTIVE: Many articles report seat belt injuries to children. This study examines the effect of child versus adult age and seat belt use on outcome in severe motor vehicle crashes. METHODS: A population-based data set of all motor vehicle crashes statewide was analyzed by using matched-pairs logistic regression. Subjects were participants in motor vehicle crashes in which at least one occupant was killed or hospitalized and at least one was a child (age <15 years). Only passengers in cars, vans, and the front of light trucks were considered. Unique matched pairs were formed of one adult and one child from the same vehicle. The main outcome measure was death or hospitalization. Covariates were seat belt use and front or back seat position. RESULTS: Overall, 413 pairs were analyzed. Seat belt use in these severe crashes was low for children and adults (40% versus 45%). Children more often sat in the back seat (74% versus 31% for adults). Risk of death was similar (7% for children and 8% for adults), but the percentage killed or hospitalized differed (13% for children and 28% for adults; odds ratio [OR] 2.5; 95% confidence interval [CI] 1.8 to 3.7). After controlling for seat belt use and seat position, adults remained at a similarly increased risk compared with children (OR 2.6; 95% CI 1.6 to 4.2). The back seat was much safer than the front seat (OR 5.5; 95% CI 3.7 to 8.1). An adult's nonuse of restraints was strongly predictive of a child's nonuse. CONCLUSION: Seat belts were at least as protective for children as for adults, but only 40% of the children in these severe crashes were restrained. PMID- 11020682 TI - Teenagers' perceptions of personal safety and the role of the emergency health care provider. AB - STUDY OBJECTIVE: We designed this study to determine the experiences, attitudes, and beliefs of teenagers regarding violence in their lives and to gain an understanding of the perceived role of the emergency health care professional. METHODS: A qualitative study involving 10-person focus groups was conducted in 4 cities representing urban/low socioeconomic and suburban/high socioeconomic areas. Participants were 14 or 15 years of age and were recruited from local community centers. Moderators were matched by sex to the teenagers, and groups were segmented by race and sex. A semistructured guide was developed to help facilitate the discussion. All groups were audiotaped and videotaped, and the tapes were reviewed by the investigators for reoccurring themes. RESULTS: A total of 140 adolescents (14 groups of 10) participated; one half were male. Urban teenagers expressed concerns about gangs, rape, and homicide; suburban teenagers were concerned about parental pressure and suicide. The teenagers expressed distrust of teachers, police officers, and doctors and felt safest with their parents. The emergency department was viewed as a confusing and frightening place, and participants believed that the role of the ED staff was to treat the patient's medical problem and not inquire or counsel about violence. CONCLUSION: All of the teenagers, regardless of socioeconomic status, were concerned about violence in their lives. All of the teenagers believed that the emergency department is not the place for patients to be counseled about safety and violence prevention. A better understanding of the problem of violence from the point of view of the teenager is important in refining an effective role for the emergency health care provider in adolescent violent injury prevention. PMID- 11020683 TI - Immobilization technique and blade choice in the endotracheal intubation of trauma patients: Miller time or much ado about nothing? PMID- 11020684 TI - Why did the chicken cross the road? ... to get the antibiotics. PMID- 11020685 TI - Design and implementation of a controlled trial of pediatric endotracheal intubation in the out-of-hospital setting. AB - This article describes the design and implementation of the Pediatric Airway Management Project. The project was completed January 1, 1997, and evaluated the effectiveness of endotracheal intubation relative to bag-valve-mask ventilation in improving survival to hospital discharge and neurologic outcome in children, the effect of training on paramedic airway management skills and self-efficacy, the length of time the skills can be retained, and the costs of training and retraining. The main focus of project design was the implementation of a controlled trial comparing methods of airway management for acutely ill and injured pediatric patients in the out-of-hospital setting. To date, this project is the largest prospective, controlled, out-of-hospital study of the care of children ever reported. Barriers to implementation of a study of this size are described. PMID- 11020686 TI - Nonoccupational HIV postexposure prophylaxis: a new role for the emergency department. AB - Despite numerous primary prevention campaigns, new cases of HIV infection are occurring at high rates. Postexposure prophylaxis (PEP) after possible HIV exposures from sexual encounters or injection drug use may prove to be a worthwhile means of reducing HIV infection. Although there are no studies that directly demonstrate its efficacy, indirect support comes from animal and human studies. Multiple animal studies have shown that antiretroviral medications can reduce simian immunodeficiency virus infections if given early and for a prolonged period. A study of health care workers suggests that zidovudine taken after needlestick injuries can dramatically reduce HIV seroconversion. Zidovudine and nevirapine use recently showed great reductions in perinatal HIV transmission. Studies of dendritic and T-cell processing of simian immunodeficiency virus and HIV indicate that antiretroviral medications taken soon after a viral exposure may terminate viral replication. Regimens of 2 or 3 antiretroviral medications have been suggested as prophylactic measures after certain exposures. Even though limited experience exists with these populations, HIV PEP is most likely safe in pregnancy and for children. Emergency departments are encouraged to anticipate the probable demands for nonoccupational HIV PEP by establishing protocols for its rapid provision and ensuring proper follow-up care. PMID- 11020687 TI - Guidelines for ambulance diversion. American College of Emergency Physicians. PMID- 11020688 TI - An international training program to assist with establishing emergency medicine in Ethiopia. AB - Ethiopia has a population of 56 million and an area of 1,110,000 km(2). Ethiopia is one of the poorest nations in the world, and its health services system reflects that poverty. Accidental injury, violence, infectious diseases, and natural and manmade disasters abound, yet emergency medicine is practically nonexistent. Poorly equipped and staff emergency "rooms" supply substandard service even in the capital. Significant work and planning are under way by a group of high-quality and dedicated local physicians and nurses with international support from Israel and the United States. A coherent plan is due at the end of the year. Much financial and professional support from outside sources will be required for significant advances in the quality of emergency care to improve. Human as well are material resources are vital. PMID- 11020689 TI - Ecthyma gangrenosum as a manifestation of Pseudomonas sepsis in a previously healthy child. AB - We present a case of Pseudomonas aeruginosa sepsis heralded by ecthyma gangrenosum in a previously healthy 15-month-old child. Pseudomonas infection and its uncommon skin manifestation are rarely encountered in an immunocompetent child. This case highlights the critical importance of identifying ecthyma gangrenosum to institute optimal antimicrobial therapy. PMID- 11020690 TI - Antihistamines and driving-related behavior: A review of the evidence for impairment. PMID- 11020691 TI - Commentary: antihistamines and driving performance-An underrecognized issue in traffic safety PMID- 11020692 TI - The value of the out-of-hospital experience for emergency medicine residents. PMID- 11020693 TI - Today I am 2 months old PMID- 11020694 TI - Remember me PMID- 11020695 TI - Failure of intravenous calcium gluconate for hydrofluoric acid burns. PMID- 11020696 TI - Ultrasound-guided brachial and basilic vein cannulation. PMID- 11020697 TI - Enhancing the emergency department outcomes of patients with mental retardation. PMID- 11020699 TI - Clinical policy: critical issues for the initial evaluation and management of patients presenting with a chief complaint of nontraumatic acute abdominal pain PMID- 11020711 TI - Nodal and bone morphogenetic protein 5 interact in murine mesoderm formation and implantation. AB - Mice mutant for the TGF-beta family member, nodal, lack mesoderm and die between E8.5 and E9.5. The short ear-lethal (se(l) ) mutation, a deletion that eliminates Bmp-5, causes a strikingly similar gastrulation defect. Here we analyze se(l);nodal compound mutants and find a dosage effect. Embryos homozygous for one mutation show distinct gastrulation stage defects that depend on whether they are heterozygous or homozygous for the other mutation. Embryos mutant for nodal or se(l);nodal compound mutants fail to execute an antigenic shift indicative of mesoderm differentiation and ectoderm cells are shunted into an apoptotic pathway. Furthermore, we find a novel phenotype in se(l);nodal double mutant litters, in which two to four genetically different embryos are contained within the same deciduum. Both the gastrulation and implantation phenotypes can also arise in short ear-viable (se(v) ) and se(v); nodal mutant mice. These data indicate that loss of Bmp-5 may underlie the se(l) gastrulation phenotype and suggest that nodal and Bmp-5 interact during murine mesoderm formation. Our data also reveal an unsuspected role for Bmp-5 in implantation and the decidual response in the mouse. PMID- 11020713 TI - A flattened mouse embryo: leveling the playing field. PMID- 11020712 TI - Temporally controlled somatic mutagenesis in smooth muscle. AB - Ligand-dependent site-specific recombinases are powerful tools to engineer the mouse genome in specific somatic cell types at selected times during pre- and postnatal development. Current efforts are primarily directed towards increasing the efficiency of this recombination system in mice. We have generated transgenic mouse lines expressing a tamoxifen-activated Cre recombinase, CreER(T2), under the control of the smooth muscle-specific SM22 promoter. Both a randomly integrated transgene [SM-CreER(T2)(tg)] and a transgene that has been "knocked in" into the endogenous SM22 locus [SM-CreER(T2)(ki)] were expressed in smooth muscle-containing tissues. The level of CreER(T2) expression and tamoxifen induced recombination was lower in SM-CreER(T2)(tg) mice compared with SM CreER(T2)(ki) mice. Whereas no recombinase activity could be detected in vehicle treated SM-CreER(T2)(ki) mice, administration of tamoxifen induced the excision of a loxP-flanked reporter transgene in up to 100% of smooth muscle cells. The recombined genome persisted for at least four months after tamoxifen treatment. SM-CreER(T2)(ki) transgenic mice should be useful to study the effects of various somatic mutations in smooth muscle. PMID- 11020714 TI - A new positive/negative selectable marker, puDeltatk, for use in embryonic stem cells. AB - A novel positive/negative selection cassette, puDeltatk, was generated. pu(Delta)tk is a bifunctional fusion protein between puromycin N acetyltransferase (Puro) and a truncated version of herpes simplex virus type 1 thymidine kinase (DeltaTk). Murine embryonic stem (ES) cells transfected with pu(Delta)tk become resistant to puromycin and sensitive to 1-(-2-deoxy-2-fluoro-1 beta-D-arabino-furanosyl)-5-iodouracil (FIAU). Unlike other HSV1 tk transgenes, puDeltatk is readily transmitted through the male germ line. Thus pu(Delta)tk is a convenient positive/negative selectable marker that can be widely used in many ES cell applications. PMID- 11020739 TI - Cause-specific mortality risks of anesthesiologists. New evidence for the existence of old problems. PMID- 11020715 TI - Genetically distinct cardial cells within the Drosophila heart. AB - Although often viewed as a simple pulsating tube, the Drosophila dorsal vessel is intricate in terms of its structure, cell types, and patterns of gene expression. Two nonidentical groups of cardial cells are observed in segments of the heart based on the differential expression of transcriptional regulators. These include sets of four cell pairs that express the homeodomain protein Tinman (Tin), alternating with groups of two cell pairs that express the orphan steroid hormone receptor Seven Up (Svp). Here we show that these myocardial cell populations are distinct in terms of their formation and gene expression profiles. The Svp expressing cells are generated by asymmetric cell divisions of precursor cells based on decreases or increases in their numbers in numb or sanpodo mutant embryos. In contrast, the numbers of Tin-expressing cardial cells are unchanged in these genetic backgrounds, suggesting they arise from symmetric cell divisions. One function for Svp in the two pairs of cardial cells is to repress the expression of the tin gene and at least one of its targets, the beta3 tubulin gene. Further differences in the cells are substantiated by the identification of separable enhancers for D-mef2 gene transcription in the distinct cardioblast sets. Taken together, these results demonstrate a greater cellular and genetic complexity of the Drosophila heart than previously appreciated. PMID- 11020740 TI - Cause-specific mortality risks of anesthesiologists. AB - BACKGROUND: The health-related effects of the operating room environment are unclear. The authors compared mortality risks of anesthesiologists to those of internal medicine physicians between 1979 and 1995. METHODS: The Physician Master File database, a listing of all US physicians, was used to identify anesthesiologists and general internists. The cohort of internists (n = 40,211) was a stratified random sample of all internists, frequency-matched to the cohort of anesthesiologists (n = 40,242) by gender, decade of birth, and US citizenship. The National Death Index was used to confirm death status and to determine specific causes of death. Mortality risks, adjusted for age, gender, and race, were compared using the Cox proportional hazards regression model. RESULTS: The standardized mortality ratios for all physicians were well below 1.0, except for suicide. The all-cause mortality ratios, and the risks of death caused by cancer and heart disease, did not differ between anesthesiologists and internists. Anesthesiologists had an increased risk of death from suicide (rate ratio [RR] = 1.45, 95% confidence interval [CI] = 1.07 - 1.97), drug-related death (RR = 2.79, 95% CI = 1.87 - 4.15), death from other external causes (RR = 1.53, 95% CI = 1.05 - 2.22), and death from cerebrovascular disease (RR = 1.39, 95% CI = 1.08 - 1.79). Male anesthesiologists had an increased risk of death from HIV (RR = 1.82, 95% CI = 1.09 - 3.02) and viral hepatitis (RR = 7.98, 95% CI = 1.0 - 63.84). Although the risk to anesthesiologists of drug-related deaths was highest in the first 5 years after medical school graduation, it remained increased over that of internists throughout the career. CONCLUSIONS: Substance abuse and suicide represent significant occupational hazards for anesthesiologists. New methods to combat substance abuse among anesthesiologists should be developed. PMID- 11020741 TI - Substance P (Neurokinin-1) antagonist prevents postoperative vomiting after abdominal hysterectomy procedures. AB - BACKGROUND: The safety and antiemetic efficacy of CP-122,721, a novel neurokinin 1 antagonist, was evaluated when administered alone or in combination with ondansetron. METHODS: Using a randomized, double-blind, placebo-controlled study design, CP-122,721 was initially compared with placebo and subsequently to ondansetron alone and in combination for prophylaxis against postoperative nausea and vomiting in 243 women undergoing abdominal hysterectomy. In the dose-ranging studies (n = 86), patients received either CP-122,721 100 mg (vs. placebo) or 200 mg (vs. placebo) orally 60-90 min before induction of anesthesia. In the interaction study (n = 157), patients received CP-122,721 200 mg or placebo 60-90 min before induction of anesthesia, and ondansetron 4 mg or saline 2 ml intravenously 15-30 min before the end of surgery. Patients assessed their level of nausea and pain on arrival in the postanesthesia care unit and at 0.5-, 1-, 1.5-, 2-, 4-, 8-, 12-, and 24-h intervals postoperatively. Emetic episodes, need for rescue antiemetic-antinausea medication, postoperative complications, and patient satisfaction were recorded. RESULTS: In the initial dose-ranging study, only 10% of the patients experienced emesis within the first 8 h after surgery with CP-122,721 200 mg compared with 50% in the placebo group. CP-122,721 200 mg also decreased the need for rescue medication (25% vs. 48%). CP-122,721 100 mg was less effective than 200 mg in decreasing the incidence of repeated episodes of emesis. In the interaction study, 6% of the patients receiving CP-122,721 200 mg orally experienced emesis less than 2 h after surgery compared with 17% with ondansetron alone. With combined therapy, only 2% experienced emesis. In addition, the median times for 75% of patients to remain free from postoperative nausea and vomiting were 82, 75, and 362 min in the ondansetron, CP-122,721, and combination groups, respectively. CONCLUSIONS: Oral CP-122,721 200 mg decreased emetic episodes compared with ondansetron (4 mg intravenously) during the first 24 h after gynecologic surgery; however, there was no difference in patient satisfaction. PMID- 11020742 TI - Lower extremity neuropathies associated with lithotomy positions. AB - BACKGROUND: The goal of this project was to study the frequency and natural history of perioperative lower extremity neuropathies. METHODS: A prospective evaluation of lower extremity neuropathies in 991 adult patients undergoing general anesthetics and surgical procedures while positioned in lithotomy was performed. Patients were assessed with use of a standard questionnaire and neurologic examination before surgery, daily during hospital stay in the first week after surgery, and by phone if discharged before 1 postoperative week. Patients in whom lower extremity neuropathies developed were observed for 6 months. RESULTS: Lower extremity neuropathies developed in 15 patients (1.5%; 95% confidence interval, 0.8-2.5%). Unilateral or bilateral nerves were affected in patients as follows: obturator (five patients), lateral femoral cutaneous (four patients), sciatic (three patients), and peroneal (three patients). Paresthesia occurred in 14 of 15 patients, and 4 patients had burning or aching pain. No patient had weakness. Symptoms were noted within 4 h of completion of the anesthetic in all 15 patients. These symptoms resolved within 6 months in 14 of 15 patients. Prolonged positioning in a lithotomy position, especially for more than 2 h, was a major risk factor for this complication (P = 0.006). CONCLUSIONS: In this surgical population, lower extremity neuropathies were infrequent complications that were noted very soon after surgery and anesthesia. None resulted in prolonged disability. The longer patients were positioned in lithotomy positions, the greater the chance of development of a neuropathy. These findings suggest that a reduction of duration of time in lithotomy positions may reduce the risk of lower extremity neuropathies. PMID- 11020743 TI - Transdermal nitroglycerine enhances spinal neostigmine postoperative analgesia following gynecological surgery. AB - BACKGROUND: Intrathecal neostigmine causes analgesia by inhibiting the breakdown of acetylcholine. Experimental data suggest that the production of endogenous nitric oxide is necessary for tonic cholinergic inhibition of spinal pain transmission. The purpose of this study was to determine whether association of transdermal nitroglycerine would enhance analgesia from a low dose of intrathecal neostigmine in patients undergoing gynecologic surgery during spinal anesthesia. METHODS: Forty-eight patients were randomized to one of four groups. Patients were premedicated with use of 0.05-0.1 mg/kg intravenous midazolam and received 15 mg bupivacaine plus 1 ml test drug intrathecally (saline or neostigmine, 5 microgram). Twenty to 30 min after the spinal puncture, a transdermal patch of either 5 mg nitroglycerin or placebo was applied. The control (Con) group received spinal saline and transdermal placebo. The neostigmine group received spinal neostigmine and transdermal placebo. The nitroglycerin group received spinal saline and a transdermal nitroglycerine patch. Finally, the neostigmine nitroglycerin group received spinal neostigmine and transdermal nitroglycerine. Pain and adverse effects were evaluated using a 10-cm visual analog scale. RESULTS: Patients in the groups were similar regarding age, weight, height, and American Society of Anesthesiologists status. Sensory level to pin prick at 10 min, surgical duration, anesthetic duration, and visual analog scale score for pain at the time of administration of first rescue medication were statistically the same for all groups. The time to administration of first rescue analgesic (min) was longer in the neostigmine-nitroglycerin group (550 min; range, 458 1,440 min; median, 25-75th percentile) compared with the other groups (P < 0.001). The neostigmine-nitroglycerin group required fewer rescue analgesics in 24 h than did the control group (P < 0.0005), whereas the neostigmine group required less analgesics compared with the control group (P < 0.02). The incidence of perioperative adverse effects (nausea, vomiting, headache, back pain) was similar among groups (P > 0.05). CONCLUSION: Although neither intrathecal 5 microgram neostigmine alone nor transdermal nitroglycerine alone (5 mg/day) delayed the time to administration of first rescue analgesics, the combination of both provided an average of 14 h of effective postoperative analgesia after vaginoplasty, suggesting that transdermal nitroglycerin and the central cholinergic agent neostigmine may enhance each other's antinociceptive effects at the dose studied. PMID- 11020744 TI - Arterial and venous contributions to near-infrared cerebral oximetry. AB - BACKGROUND: Cerebral oximetry is a noninvasive bedside technology using near infrared light to monitor cerebral oxygen saturation (Sco2) in an uncertain mixture of arteries, capillaries, and veins. The present study used frequency domain near-infrared spectroscopy to determine the ratio of arterial and venous blood monitored by cerebral oximetry during normoxia, hypoxia, and hypocapnia. METHODS: Twenty anesthetized children aged < 8 yr with congenital heart disease of varying arterial oxygen saturation (Sao2) were studied during cardiac catheterization. Sco2, Sao2, and jugular bulb oxygen saturation (Sjo2) were measured by frequency domain near-infrared spectroscopy and blood oximetry at normocapnia room air, normocapnia 100% inspired O2, and hypocapnia room air. RESULTS: Among subject conditions, Sao2 ranged from 68% to 100%, Sjo2 from 27% to 96%, and Sco2 from 29% to 92%. Sco2 was significantly related to Sao2 (y = 0. 85 x -17, r = 0.47), Sjo2 (y = 0.77 x +13, r = 0.70), and the combination (Sco2 = 0.46 Sao2 + 0.56 Sjo2 - 17, R = 0.71). The arterial and venous contribution to cerebral oximetry was 16 +/- 21% and 84 +/- 21%, respectively (where Sco2 = alpha Sao2 + beta Sjo2 with alpha and beta being arterial and venous contributions). The contribution was similar among conditions but differed significantly among subjects (range, approximately 40:60 to approximately 0:100, arterial:venous). CONCLUSIONS: Cerebral oximetry monitors an arterial/venous ratio of 16:84, similar in normoxia, hypoxia, and hypocapnia. Because of biologic variation in cerebral arterial/venous ratios, use of a fixed ratio is not a good method to validate the technology. PMID- 11020745 TI - Oral clonidine premedication does not change efficacy of simulated epidural test dose in sevoflurane-anesthetized children. AB - BACKGROUND: Caudal epidural anesthesia is often used as an adjunct to general anesthesia and for postoperative pain relief in children. In anesthetized children, epinephrine and isoproterenol are reliable indicators to detect accidental intravascular injection of a test dose. Oral clonidine, a useful premedicant in pediatric anesthesia, modifies hemodynamic responses to sympathomimetics, including catecholamines. The aim of the current study was to determine whether oral clonidine premedication alters the efficacy of a simulated intravascular test dose containing epinephrine or isoproterenol in sevoflurane anesthetized children. METHODS: One hundred twenty children (aged 1-7 yr) were randomly divided into six groups; control-saline, control-epinephrine, control isoproterenol, clonidine-saline, clonidine-epinephrine, and clonidine isoproterenol. The three clonidine groups received oral clonidine 4 microg/kg [DOSAGE ERROR CORRECTED] as premedication, whereas the three control groups did not receive any premedication. Anesthesia was maintained with sevoflurane at a level of 1.2 minimum alveolar concentration. After hemodynamics were stable, 0.1 ml/kg of 1% lidocaine containing epinephrine 0.5 mg/kg or isoproterenol 75 ng/kg was intravenously given to the two epinephrine or isoproterenol groups, respectively, to simulate intravascular injection of a test dose. The saline groups received saline alone instead of the test dose. Heart rate, blood pressure, and T-wave amplitude of electrocardiogram were recorded before and after administration of study drugs for subsequent analysis. RESULTS: Test solution containing epinephrine increased heart rate, systolic blood pressure, and T-wave amplitude. Oral clonidine had no effect on elevation of these variables in response to epinephrine. The isoproterenol-containing test dose produced a prominent increase in heart rate and a less pronounced increase in systolic blood pressure and T-wave amplitude. Oral clonidine also failed to modify isoproterenol-induced hemodynamic and T-wave changes. Calculated sensitivity and specificity of epinephrine or isoproterenol were all 100% based on a new heart rate criterion (positive if >/= 10 beats/min) and were unaltered by oral clonidine premedication. CONCLUSIONS: Epinephrine or isoproterenol is a reliable marker to detect accidental intravascular injection of a test dose with 100% sensitivity and specificity based on a new heart rate criterion in sevoflurane-anesthetized children. These data suggest that oral clonidine premedication does not alter the efficacy of a simulated epidural test dose containing epinephrine or isoproterenol. PMID- 11020746 TI - Emergence from anesthesia in the prone versus supine position in patients undergoing lumbar surgery. AB - BACKGROUND: Conventional supine emergence in patients undergoing prone lumbar surgery frequently results in tachycardia, hypertension, coughing, and loss of monitoring as the patient is rolled supine. The prone position might facilitate a smoother emergence because the patient is not disturbed. No data describe this technique. METHODS: Fifty patients were anesthetized with fentanyl, nitrous oxide, isoflurane, and rocuronium. By the conclusion of surgery, all patients achieved spontaneous ventilation and full reversal of neuromuscular blockade in the prone position, as the volatile anesthetic level was reduced. Baseline heart rate and mean arterial pressure were recorded. Patients were then randomized at time 0 to the supine (n = 24) or prone (n = 21) position as 100% oxygen was administered. Patients in the supine position were then rolled over, while those in the prone position remained undisturbed. Heart rate, mean arterial pressure, and coughs were recorded until extubation. Tracheas were extubated on eye opening or purposeful behavior. RESULTS: When compared with the supine group, prone patients had significantly less increase in heart rate (P = 0.0003, maximum increase 9.3 vs. 25 beats/min), less increase in mean arterial pressure (P = 0.0063, maximum increase 4.8 vs. 19 mmHg), less coughing (P = 0.0004, 7.0 vs. 23 coughs), and fewer monitor disconnections (P < 0.0001). Time to extubation from time 0 was similar (4.0 vs. 3.7 min, prone vs. supine). No one required airway rescue. There was no significant difference in need for restraint (three prone, four supine). CONCLUSIONS: Prone emergence and extubation is associated with less hemodynamic stimulation, less coughing, and less disruption of monitors, without specifically observed adverse effects, when compared with conventional supine techniques. PMID- 11020747 TI - Evaluation of a cerebral oximeter as a monitor of cerebral ischemia during carotid endarterectomy. AB - BACKGROUND: Stroke is an important contributor to perioperative morbidity and mortality associated with carotid endarterectomy (CEA). This investigation was designed to compare the performance of the INVOS-3100 cerebral oximeter to neurologic function, as a means of detecting cerebral ischemia induced by carotid cross-clamping, in patients undergoing carotid endarterectomy with cervical plexus block. METHODS: Ninety-nine patients undergoing 100 CEAs with regional anesthesia (deep or superficial cervical plexus block) were studied. Bilateral regional cerebrovascular oxygen saturation (rSO2) was monitored using the INVOS 3100 cerebral oximeter. Patients were retrospectively assigned to one of two groups: those in whom a change in mental status or contralateral motor deficit was noted after internal carotid clamping (neurologic symptoms; n = 10) and those who did not show any neurologic change (no neurologic symptoms; n = 90). Data from 94 operations (neurologic symptoms = 10 and no neurologic symptoms = 84) were adequate for statistical analyses for group comparisons. A relative decrease in ipsilateral rSO2 after carotid occlusion (calculated as a percentage of preocclusion value) during all operations (n = 100) was also calculated to determine the critical level of rSO2 decrease associated with a change in neurologic function. RESULTS: The mean (+/- SD) decrease in rSO2 after carotid occlusion in the neurologic symptoms group (from 63.2 +/- 8.4% to 51.0 +/- 11.6%) was significantly greater (P = 0.0002) than in the no neurologic symptoms group (from 65.8 +/- 8.5% to 61.0 +/- 9.3%). Logistic regression analysis used to determine if a change in rSO2, calculated as a percentage of preclamp value, could be used to predict change in neurologic function was highly significant (likelihood ratio chi-square = 13.7; P = 0.0002). A 20% decrease in rSO2 reading from the preclamp baseline, as a predictor of neurologic compromise, resulted in a sensitivity of 80% and specificity of 82.2%. The false-positive rate using this cutoff point was 66.7%, and the false-negative rate was 2.6%, providing a positive predictive value of 33.3% and a negative predictive value of 97.4%. CONCLUSION: Monitoring rSO2 with INVOS-3100 to detect cerebral ischemia during CEA has a high negative predictive value, but the positive predictive value is low. PMID- 11020748 TI - Transcranial Doppler ultrasonography as a screening technique for detection of a patent foramen ovale before surgery in the sitting position. AB - BACKGROUND: Venous air embolism has been reported to occur in 23-45% of patients undergoing neurosurgical procedures in the sitting position. If venous air embolism occurs, a patent foramen ovale (PFO) is a risk factor for paradoxical air embolism and its sequelae. Preoperative screening for a PFO is therefore recommended by some investigators. The reference standard for identifying a PFO is contrast-enhanced transesophageal echocardiography (c-TEE). Contrast-enhanced transcranial Doppler ultrasonography (c-TCD) and contrast-enhanced transthoracic echocardiography (c-TTE) are noninvasive alternative methods, but so far there are no studies as to their diagnostic validity in neurosurgical patients. METHODS: The sensitivity and specificity of c-TCD and c-TTE in detecting a PFO were determined in a prospective study using c-TEE as the reference standard. Preoperative c-TCD, c-TTE, and c-TEE studies were performed during the Valsalva maneuver after intravenous echo-contrast medium (D-Galactose, Echovist-300, Schering AG, Berlin, Germany) was administered in 92 consecutive candidates (47 men and 45 women; mean age, 51 yr; range, 25-72 yr) before neurosurgical procedures in the sitting position. RESULTS: A PFO was detected in 24 of the 92 patients (26.0%) using c-TEE. c-TCD correctly identified 22 patients, whereas c TTE only correctly identified 10. This corresponds to a sensitivity of 0.92 for c TCD and 0.42 for c-TTE. The negative predictive value was 0.97 for c-TCD compared with 0.83 for c-TTE. The prevalence of a PFO in patients with a posterior fossa lesion was 27%, and in the group with cervical disc herniation was 24% as detected by c-TEE. The incidence of intraoperative venous air embolism was 35% in cases of cervical foraminotomy and 75% in posterior fossa surgery as detected by c-TEE. CONCLUSIONS: c-TCD is a highly sensitive and highly specific method for detecting a PFO. Because c-TCD is noninvasive, it may be more suitable than c-TEE for routine preoperative screening for a PFO. C-TTE is not reliable in detecting a PFO. PMID- 11020749 TI - Analgesic effects of caudal and intramuscular S(+)-ketamine in children. AB - BACKGROUND: Previous studies suggest that caudal administration of ketamine cause effective analgesia. The purpose of the current study was to compare the clinical effectiveness and plasma concentrations of S(+)-ketamine after caudal or intramuscular administration in children to distinguish between local and systemic analgesia. METHODS: After induction of general anesthesia, 42 patients, aged 1 to 7 yr, scheduled to undergo inguinal hernia repair randomly received a caudal (caudal group) or intramuscular (intramuscular group) injection of 1 mg/kg S(+)-ketamine. Intraoperatively, heart rate (HR), mean arterial pressure (MAP) and arterial oxygen saturation were measured. Postoperative measurements included duration of analgesia, a four-point sedation score, and hemodynamic and respiratory monitoring for 6 h in the recovery room. Analgesic requirements in the recovery room were assessed by an independent blinded observer using an observational pain/discomfort scale (OPS). Plasma samples for determination of ketamine concentrations were obtained before and 10, 20, 30, 45, 60, 90, 120, and 180 min after injection of S(+)-ketamine. RESULTS: A significantly longer duration of analgesia (P < 0.001) was observed after caudal administration (528 min [220-1,440 min]; median [range]) when compared with intramuscular administration (108 min [62-1,440 min]) of S(+)-ketamine. Plasma levels of ketamine were significantly lower from 10 to 45 min after caudal administration than after intramuscular injection. CONCLUSION: Caudal S(+)-ketamine provides good intra- and postoperative analgesia in children. Despite similar plasma concentrations during most of the postoperative observation period, caudal S(+) ketamine provided more effective analgesia than did intramuscular S(+)-ketamine, indicating a local analgesic effect. PMID- 11020750 TI - Electroencephalogram approximate entropy correctly classifies the occurrence of burst suppression pattern as increasing anesthetic drug effect. AB - BACKGROUND: Approximate entropy, a measure of signal complexity and regularity, quantifies electroencephalogram changes during anesthesia. With increasing doses of anesthetics, burst-suppression patterns occur. Because of the high-frequency bursts, spectrally based parameters such as median electroencephalogram frequency and spectral edge frequency 95 do not decrease, incorrectly suggesting lightening of anesthesia. The authors investigated whether the approximate entropy algorithm correctly classifies the occurrence of burst suppression as deepening of anesthesia. METHODS: Eleven female patients scheduled for elective major surgery were studied. After propofol induction, anesthesia was maintained with isoflurane only. Before surgery, the end-tidal isoflurane concentration was varied between 0.6 and 1.3 minimum alveolar concentration. The raw electroencephalogram was continuously recorded and sampled at 128 Hz. Approximate entropy, electroencephalogram median frequency, spectral edge frequency 95, burst suppression ratio, and burst-compensated spectral edge frequency 95 were calculated offline from 8-s epochs. The relation between burst-suppression ratio and approximate entropy, electroencephalogram median frequency, spectral edge frequency 95, and burst-compensated spectral edge frequency 95 was analyzed using Pearson correlation coefficient. RESULTS: Higher isoflurane concentrations were associated with higher burst-suppression ratios. Electroencephalogram median frequency (r = 0.34) and spectral edge frequency 95 (r = 0.29) increased, approximate entropy (r = -0.94) and burst-compensated spectral edge frequency 95 (r = -0.88) decreased with increasing burst-suppression ratio. CONCLUSION: Electroencephalogram approximate entropy, but not electroencephalogram median frequency or spectral edge frequency 95 without burst compensation, correctly classifies the occurrence of burst-suppression pattern as increasing anesthetic drug effect. PMID- 11020751 TI - Pulmonary disposition of propofol in surgical patients. AB - BACKGROUND: The lungs have been mentioned as a possible site contributing to the extrahepatic clearance of propofol. The objective of the present study was to clarify the pulmonary disposition of propofol directly in human lungs by investigating both the first-pass uptake and pulmonary extraction at pseudo steady state. METHODS: Nine patients were enrolled in the first-pass uptake study. Propofol (5 mg) and indocyanine green (ICG; 15 mg) were simultaneously administered via a central venous catheter within 1 s, and sequential arterial blood samples were obtained from the radial artery at 1-s intervals up to 45 s. Eleven patients were included in the infusion study, and propofol was infused via the jugular vein at a rate of 50 microgram. kg-1. min-1. Blood samples were simultaneously collected from pulmonary and radial arteries up to 60 min. RESULTS: A pronounced difference in the dilution curves between propofol and ICG was observed, and 28.4 +/- 11.6% (mean +/- SD) of propofol was taken up during the single passage through the human lung. The mean pulmonary transit time of propofol (31.3 +/- 6.0 s) was significantly longer than that of ICG (22.4 +/- 2.7 s; P < 0.01), indicating that some of the propofol trapped by lungs returned to the circulation by back diffusion. In the constant infusion study, no significant differences were observed with the plasma concentrations of propofol between pulmonary and radial arteries except for that at 2 min. The area under the curve of pulmonary and radial arterial concentration curves to 60 min were 59.1 +/- 14.8 and 56.8 +/- 12.5 microg. ml-1. min-1, respectively. No significant difference was observed with the area under the curve, suggesting that metabolism was not involved in the pulmonary uptake in human lungs. CONCLUSIONS: Most of the propofol that undergoes pulmonary uptake during the first pass was released back to the circulation by back diffusion. Metabolism was not involved in the pulmonary uptake in human lungs. PMID- 11020752 TI - The role of human lungs in the biotransformation of propofol. AB - BACKGROUND: The metabolism of propofol is very rapid, and its transformation takes place mainly in the liver. There are reports indicating extrahepatic metabolism of the drug, and the alimentary canal, kidneys, and lungs are mentioned as the most probable places where the process occurs. The aim of this study was to determine whether the human lungs really take part in the process of propofol biotransformation. METHODS: Blood samples were taken from 55 patients of American Society of Anesthesiologists grade 1-3 scheduled for elective intracranial procedures (n = 47) or for pulmonectomy (n = 8). All patients were premedicated with diazepam (10 mg) administered orally 2 h before anesthesia. Propofol total intravenous anesthesia was performed at the following infusion rates: 12 mg. kg-1. h-1, 9 mg. kg-1. h-1, and 6 mg. kg-1. h-1. Fentanyl and pancuronium bromide were also administered intermittently. After tracheal intubation, the lungs were ventilated to normocapnia with an oxygen-air mixture (fraction of inspired oxygen = 0.33). Blood samples for propofol and 2,6 diisopropyl-1, 4-quinol analysis were taken simultaneously from the right atrium and the radial artery, or the pulmonary artery and the radial artery. The concentration of both substances were measured with high-performance liquid chromatography and gas chromatography-mass spectroscopy. RESULTS: The concentration of propofol in the central venous system (right atrium or pulmonary artery) is greater than in the radial artery, whereas the opposite is observed for propofol's metabolite, 2,6-diisopropyl-1,4-quinol. Higher propofol concentrations are found in blood taken from the pulmonary artery than in the blood collected from the radial artery. CONCLUSIONS: Human lungs take part in the elimination of propofol by transforming the drug into 2,6-diisopropyl-1,4-quinol. PMID- 11020753 TI - Adenosine-induced ventricular asystole to induce transient profound systemic hypotension in patients undergoing endovascular therapy. Dose-response characteristics. AB - BACKGROUND: Adenosine-induced asystole has been used to induce transient systemic hypotension for various vascular procedures. Dose-response characteristics of adenosine-induced ventricular asystole have not been determined. METHODS: During endovascular embolization of cerebral arteriovenous malformations, the authors performed a series of adenosine test injections to establish a dose-response relation in each patient. After an interval of 3-10 min, the dose was escalated by 10-20 mg for each injection to achieve an end point of 20-30 s of stable mean arterial pressure (MAP) reduction to 25-30 mmHg. All patients received constant infusion of nitroprusside (approximately 1 microgram. kg-1. min-1) throughout the procedure. RESULTS: The authors studied four adult patients (age, 22-44 yr; two patients had two separate procedures) and one pediatric patient (age, 4 yr). Twenty-three adenosine injections resulted in measurable asystole. The adenosine dose was 0. 98 +/- 0.40 mg/kg (mean +/- SD), and the dose range was 0.24-1.76 mg/kg (6-90 mg). The duration of asystole, MAP < 30 mmHg, and MAP < 50 mmHg, were 8 +/- 3 s, 18 +/- 12 s, and 50 +/- 29 s, respectively. The minimum MAP and the MAP for the first 20 s were 16 +/- 3 mmHg and 30 +/- 9 mmHg, respectively. There was a linear relation between adenosine dose and the duration of hypotension with MAP < 30 mmHg and MAP < 50 mmHg. CONCLUSIONS: In the dose range studied, a series of adenosine test injections can be used to determine optimal adenosine dose for induction of transient profound hypotension. PMID- 11020754 TI - Isosulfan blue affects pulse oximetry. AB - BACKGROUND: Certain vital dyes are known to cause pulse oximetry (Spo2) desaturation. The authors studied the effect of isosulfan blue (IB) on Spo2. METHODS: Thirty-three women, aged 34-81 yr, who were undergoing surgery for breast cancer were studied. IB, 5 ml (50 mg), was injected intraparenchymally around the tumor area by the surgeon. A pulse oximeter was used to continuously record Spo2 values up to 130 min after IB injection. Friedman repeated-measures analysis of ranks was used to analyze the baseline Spo2 and values at 5, 10, 20, 30, 40, 50, and 60 min. RESULTS: Spo2 values were significantly different from baseline values at 5, 10, 20, 30, 40, 50, and 60 min (P < 0.05). In a typical patient, a maximum Spo2 decrease of 3% can be anticipated 25 min after injection of IB. CONCLUSIONS: After peritumoral administration of IB, 50 mg, a significant interference with Spo2 will occur. PMID- 11020755 TI - Electrocardiographic ST-segment changes during acute, severe isovolemic hemodilution in humans. AB - BACKGROUND: Controversy exists regarding the lowest blood hemoglobin concentration that can be safely tolerated. The authors studied healthy resting humans to test the hypothesis that acute isovolemic reduction of blood hemoglobin concentration to 5 g/dl would produce an imbalance in myocardial oxygen supply and demand, resulting in myocardial ischemia. METHODS: Fifty-five conscious healthy human volunteers were studied. Isovolemic removal of aliquots of blood reduced blood hemoglobin concentration from 12.8 +/- 1.2 to 5.2 +/- 0.5 g/dl (mean +/- SD). Removed blood was replaced simultaneously with intravenous fluids to maintain constant isovolemia. Hemodynamics and arterial oxygen content (Cao2) were measured before and after removal of each aliquot of blood. Electrocardiographic (ECG) changes were monitored continuously using a Holter ECG recorder for detection of myocardial ischemia. RESULTS: During hemodilution, transient, reversible ST-segment depression developed in three subjects as seen on the electrocardiogram during hemodilution. These changes occurred at hemoglobin concentrations of 5-7 g/dl while the subjects were asymptomatic. Two of three subjects with ECG changes had significantly higher heart rates than those without ECG changes at the same hemoglobin concentrations. When evaluating the entire study period, the subjects who had ECG ST-segment changes had significantly higher maximum heart rates than those without ECG changes, despite having similar baseline values. CONCLUSION: With acute reduction of hemoglobin concentration to 5 g/dl, ECG ST-segment changes developed in 3 of 55 healthy conscious adults and were suggestive of, but not conclusive for, myocardial ischemia. The higher heart rates that developed during hemodilution may have contributed to the development of an imbalance between myocardial supply and demand resulting in ECG evidence of myocardial ischemia. However, these ECG changes appear to be benign because they were reversible and not accompanied by symptoms. PMID- 11020756 TI - Cardiovascular and metabolic response to acute normovolemic anemia. Effects of anesthesia. AB - BACKGROUND: The maintenance of adequate tissue oxygenation during acute anemia depends on an increase in both cardiac output and tissue oxygen extraction. This study tested the hypothesis that anesthesia blunts the cardiac output response associated with acute normovolemic hemodilution. METHODS: Forty patients undergoing major abdominal surgery were prospectively randomized to undergo acute normovolemic hemodilution (ANH) either awake (awake group, n = 20) or with fentanyl-nitrous oxide-isoflurane anesthesia (anesthetized group, n = 20). Radial and pulmonary artery catheters were placed in all patients. After hemodynamic measurements were taken, patients in the two groups underwent hemodilution to decrease their hemoglobin concentration from 13 to 8 g/dl. A total of 1,875 +/- 222 ml (mean +/- SD) of blood was collected and simultaneously replaced by the same volume of medium molecular weight hydroxyethylstarch in both groups. RESULTS: In the awake group, ANH resulted in a significant increase in cardiac index (from 3.1 +/- 0.5 to 4.8 +/- 1.0 l. min-1. m-2) related to both an increase in heart rate and stroke index. Oxygen delivery remained unchanged, but oxygen consumption increased significantly, resulting in an increase in oxygen extraction ratio. In the anesthetized group, ANH resulted in a significantly smaller increase in cardiac index (from 2.3 +/- 0.5 to 3.1 +/- 0.7 l. min-1. m-2) related solely to an increase in stroke index. Oxygen delivery decreased but oxygen consumption was maintained as oxygen extraction increased. CONCLUSIONS: Anesthesia significantly reduces the cardiac output response associated with ANH. This could be related to the effects of the anesthetic drugs on the autonomic and the cardiovascular systems. PMID- 11020757 TI - Dose-response and onset/offset characteristics of rapacuronium. AB - BACKGROUND: A rigorous study of the dose-response relation of rapacuronium has, to our knowledge, yet to be performed. In addition, there is little information available regarding the onset or offset profile of rapacuronium when administered in subparalyzing doses. These issues necessitate further study. METHODS: Forty seven adult patients, American Society Anesthesiologists physical status I or II, were studied. Tracheal intubation was accomplished without muscle relaxants. Anesthesia was maintained with use of nitrous oxide, propofol, and alfentanil. The electromyogram of the first dorsal interosseous muscle was measured using a monitor. Single stimuli at 0.10 Hz were administered. A single dose of rapacuronium was administered. After log-dose or logit transformation of the data, the best-fit line of regression was determined using the method of least squares. For each subject, the authors estimated the 50% effective dose (ED50) and 95% effective dose (ED95) from the Hill equation using the slope obtained from regression analysis. The onset times to 50 and 90% of peak effect were estimated in a subset of 10 individuals in which peak twitch depression decreased to the range of 90-99%. RESULTS: The calculated ED50 and ED95 values for rapacuronium were 0.39 +/- 0.08 (SD) and 0.75 +/- 0.16 mg/kg, respectively. After a single ED95 dose, 90% of the drug's peak effect was evident in 77 +/- 17 s. After this dose, rapacuronium has a clinical duration of 6.1 +/- 1.1 min. CONCLUSIONS: The authors found the ED95 of rapacuronium to be substantially less than suggested by previous estimates. Rapacuronium has an onset profile that is not different from that previously reported for succinylcholine. The rate of spontaneous recovery was faster after rapacuronium than the authors previously observed after mivacurium administration but was slower than after succinylcholine, using an identical protocol. PMID- 11020758 TI - Block of human heart hH1 sodium channels by the enantiomers of bupivacaine. AB - BACKGROUND: S(-)-bupivacaine reportedly exhibits lower cardiotoxicity but similar local anesthetic potency compared with R(+)-bupivacaine. The bupivacaine binding site in human heart (hH1) Na+ channels has not been studied to date. The authors investigated the interaction of bupivacaine enantiomers with hH1 Na+ channels, assessed the contribution of putatively relevant residues to binding, and compared the intrinsic affinities to another isoform, the rat skeletal muscle (mu1) Na+ channel. METHODS: Human heart and mu1 Na+ channel alpha subunits were transiently expressed in HEK293t cells and investigated during whole cell voltage clamp conditions. Using site-directed mutagenesis, the authors created point mutations at positions hH1-F1760, hH1-N1765, hH1-Y1767, and hH1-N406 by introducing the positively charged lysine (K) or the negatively charged aspartic acid (D) and studied their influence on state-dependent block by bupivacaine enantiomers. RESULTS: Inactivated hH1 Na+ channels displayed a weak stereoselectivity with a stereopotency ratio (+/-) of 1.5. In mutations hH1 F1760K and hH1-N1765K, bupivacaine affinity of inactivated channels was reduced by approximately 20- to 40-fold, in mutation hH1-N406K by approximately sevenfold, and in mutations hH1-Y1767K and hH1-Y1767D by approximately twofold to threefold. Changes in recovery of inactivated mutant channels from block paralleled those of inactivated channel affinity. Inactivated hH1 Na+ channels exhibited a slightly higher intrinsic affinity than mu1 Na+ channels. CONCLUSIONS: Differences in bupivacaine stereoselectivity and intrinsic affinity between hH1 and mu1 Na+ channels are small and most likely of minor clinical relevance. Amino acid residues in positions hH1-F1760, hH1-N1765, and hH1-N406 may contribute to binding of bupivacaine enantiomers in hH1 Na+ channels, whereas the role of hH1-Y1767 remains unclear. PMID- 11020759 TI - Effects of isoflurane, sevoflurane, and halothane on myofilament Ca2+ sensitivity and sarcoplasmic reticulum Ca2+ release in rat ventricular myocytes. AB - BACKGROUND: The aim of this study was to describe and compare the effects of isoflurane, sevoflurane, and halothane at selected concentrations (i.e., concentrations that led to equivalent depression of the electrically evoked Ca2+ transient) on myofilament Ca2+ sensitivity, sarcoplasmic reticulum (SR) Ca2+ content, and the fraction of SR Ca2+ released during electrical stimulation (fractional release) in rat ventricular myocytes. METHODS: Single rat ventricular myocytes loaded with fura-2 were electrically stimulated at 1 Hz, and the Ca2+ transients and contractions were recorded optically. Cells were exposed to each anesthetic for 1 min. Changes in myofilament Ca2+ sensitivity were assessed by comparing the changes in the Ca2+ transient and contraction during exposure to anesthetic and low Ca2+. SR Ca2+ content was assessed by exposure to 20 mm caffeine. RESULTS: Isoflurane and halothane caused a depression of myofilament Ca2+ sensitivity, unlike sevoflurane, which had no effect on myofilament Ca2+ sensitivity. All three anesthetics decreased the electrically stimulated Ca2+ transient. SR Ca2+ content was reduced by both isoflurane and halothane but was unchanged by sevoflurane. Fractional release was reduced by both isoflurane and sevoflurane, but was unchanged by halothane. CONCLUSIONS: Depressed myofilament Ca2+ sensitivity contributes to the negative inotropic effects of isoflurane and halothane but not sevoflurane. The decrease in the Ca2+ transient is either responsible for or contributory to the negative inotropic effects of all three anesthetics and is either primarily the result of a decrease in fractional release (isoflurane and sevoflurane) or primarily the result of a decrease in SR Ca2+ content (halothane). PMID- 11020760 TI - Addition of sodium bicarbonate to lidocaine decreases the duration of peripheral nerve block in the rat. AB - BACKGROUND: Adding sodium bicarbonate to lidocaine to enhance its efficacy during peripheral nerve block is controversial. The authors studied the effect of adding sodium bicarbonate to lidocaine with and without epinephrine versus equivalent alkalinization by sodium hydroxide (NaOH) on onset, degree, and duration of peripheral nerve block. METHODS: Part I examined alkalinization by sodium bicarbonate versus NaOH to pH 7.8 on 0.5% lidocaine, with and without epinephrine (1:100,000), prepared from crystalline salt. Part II examined 0.5% and 1.0% commercial lidocaine solutions, with and without epinephrine, either unalkalinized or alkalinized with sodium bicarbonate or NaOH. With NaOH, pH was adjusted to 7.8, but with sodium bicarbonate, no pH adjustments were made to simulate clinical conditions. RESULTS: In part I, addition of either NaOH or sodium bicarbonate to 0.5% lidocaine without epinephrine produced a faster onset than did unalkalinized lidocaine, without effecting degree or duration of block. In solutions with epinephrine there were no differences in onset, degree, or duration between lidocaine alkalinized with sodium bicarbonate versus NaOH. In part II, addition of sodium bicarbonate or NaOH to 1.0% commercial lidocaine without epinephrine did not accelerate onset compared with the unalkalinized solution. However, adding sodium bicarbonate decreased the degree and duration of block by 25% and more than 50%, respectively, compared with lidocaine unalkalinized and alkalinized with NaOH. With epinephrine, sodium bicarbonate hastened onset without effecting degree and duration compared with the unalkalinized solution. CONCLUSIONS: With 1% commercial lidocaine without epinephrine, sodium bicarbonate decreases the degree and duration of block. However, in solutions with epinephrine, sodium bicarbonate hastens onset, without effecting degree or duration. PMID- 11020762 TI - In vitro compound A formation in a computer-controlled closed-circuit anesthetic apparatus. Comparison with a classical valve circuit. AB - BACKGROUND: Few data exist on compound A during sevoflurane anesthesia when using closed-circuit conditions and sodalime with modern computer-controlled liquid injection. METHODS: A PhysioFlex apparatus (Drager, Lubeck, Germany) was connected to an artificial test lung (inflow approximately 160 ml/min carbon dioxide, outflow approximately 200 ml/min, simulating oxygen consumption). Ventilation was set to obtain an end-tidal carbon dioxide partial pressure (Petco2) approximately 40 mmHg. Canister inflow (T degrees in) and outflow (T degrees out) temperatures were measured. Fresh sodalime and charcoal were used. After baseline analysis, sevoflurane concentration was set at 2.1% end-tidal for 120 min. At baseline and at regular intervals thereafter, Petco2, end-tidal sevoflurane, T degrees in, and T degrees out were measured. For inspiratory and expiratory compound A determination, samples of 2-ml gas were taken. These data were compared with those of a classical valve-containing closed-circuit machine. Ten runs were performed in each set-up. RESULTS: Inspired compound A concentrations increased from undetectable to peak at 6.0 (SD 1.3) and 14.3 (SD 2.5) ppm (P < 0.05), and maximal temperature in the upper outflow part of the absorbent canister was 24.3 degrees C (SD 3.6) and 39.8 degrees C (SD 1.2) (P < 0.05) in the PhysioFlex and valve circuit machines, respectively. Differences between the two machines in compound A concentrations and absorbent canister temperature at the inflow and outflow regions were significantly different (P < 0.05) at all times after 5 min. CONCLUSION: Compound A concentrations in the high flow (70 l/min), closed-circuit PhysioFlex machine were significantly lower than in conventional, valve-based machines during closed-circuit conditions. Lower absorbent temperatures, resulting from the high flow, appear to account for the lower compound A formation. PMID- 11020761 TI - Activation of mu- and delta-opioid receptors causes presynaptic inhibition of glutamatergic excitation in neocortical neurons. AB - BACKGROUND: The mechanism underlying the depressant effect of opioids on neuronal activity within the neocortex is still not clear. Three modes of action have been suggested: (1) inhibition by activation of postsynaptic potassium channels, (2) interaction with postsynaptic glutamate receptors, and (3) presynaptic inhibition of glutamate release. To address this issue, the authors investigated the effects of mu- and delta-receptor agonists on excitatory postsynaptic currents (EPSCs) and on membrane properties of neocortical neurons. METHODS: Intracellular recordings were performed in rat brain slices. Stimulus-evoked EPSCs mediated by different glutamate receptor subtypes were pharmacologically isolated, and opioids were applied by addition to the bathing medium. Possible postsynaptic interactions between glutamate and opioid receptors were investigated using microiontophoretic application of glutamate on neurons functionally isolated from presynaptic input. RESULTS: delta-Receptor activation by d-Ala2-d-Leu5-enkephalin (DADLE) reduced the amplitudes of EPSCs by maximum 60% in a naltrindole reversible manner (EC50: 6-15 nm). In 30-40% of the neurons investigated, higher concentrations (0.1-1 micrometer) of DADLE activated small outward currents. The mu-receptor selective agonist d-Ala2-N-MePhe5-Gly5-ol-enkephalin (0.1-1 micrometer) depressed the amplitudes of EPSCs by maximum 30% without changes in postsynaptic membrane properties. In the absence of synaptic transmission, inward currents induced by microiontophoretic application of glutamate were not affected by DADLE. CONCLUSIONS: Activation of mu- and delta-opioid receptors depresses glutamatergic excitatory transmission evoked in neocortical neurons by presynaptic inhibition. A weak activation of a postsynaptic potassium conductance becomes evident only at high agonist concentrations. There is no evidence for a postsynaptic interaction between glutamate and opioid receptors. PMID- 11020763 TI - Disposition of bupivacaine and its metabolites in the maternal, placental, and fetal compartments in rats. AB - BACKGROUND: This study was designed to determine the disposition of bupivacaine and its metabolites in the maternal, placental, and fetal compartments, using multiple sampling time points in chronically prepared awake pregnant rats. METHODS: All animals received an intravenous infusion of bupivacaine at a rate of 0.33 mg. kg-1. min-1 over a period of 15 min. The fetuses were delivered either at the end of infusion or at 2 or 4 h after dosing. Maternal and fetal blood and tissue samples were obtained for the assays of bupivacaine and its metabolites using capillary gas chromatography-mass spectrometry. RESULTS: The elimination half-life of bupivacaine was 37.7 min. The major metabolite was 3' hydroxybupivacaine. Bupivacaine and 3'-hydroxybupivacaine were present in all samples at the end of administration. The fetal to maternal concentration ratio of bupivacaine in plasma was 0.29, and in the placenta was 0.63. The amnion contained the highest bupivacaine concentration: threefold higher in the maternal and 11-fold higher than in the fetal plasma. At 4 h after dosing, bupivacaine was no longer detectable in any maternal and fetal samples, whereas 3' hydroxybupivacaine was still present in all tissues except the fetal plasma and heart. CONCLUSIONS: These data indicate that a considerable amount of bupivacaine is taken up by both sides of the placenta, as well as the amnion and myometrium. 3'-Hydroxybupivacaine was present in all tissues except the fetal plasma and heart samples, even after the parent compound became no longer detectable. Whether this slow elimination of 3'-hydroxybupivacaine causes any adverse effects on the fetus-newborn needs to be explored. PMID- 11020764 TI - Enflurane directly depresses glutamate AMPA and NMDA currents in mouse spinal cord motor neurons independent of actions on GABAA or glycine receptors. AB - BACKGROUND: The spinal cord is an important anatomic site at which volatile agents act to prevent movement in response to a noxious stimulus. This study was designed to test the hypothesis that enflurane acts directly on motor neurons to inhibit excitatory synaptic transmission at glutamate receptors. METHODS: Whole cell recordings were made in visually identified motor neurons in spinal cord slices from 1- to 4-day-old mice. Excitatory postsynaptic currents (EPSCs) or potentials (EPSPs) were evoked by electrical stimulation of the dorsal root entry area or dorsal horn. The EPSCs were isolated pharmacologically into glutamate N methyl-d-aspartate (NMDA) receptor- and non-NMDA receptor-mediated components by using selective antagonists. Currents also were evoked by brief pulse pressure ejection of glutamate under various conditions of pharmacologic blockade. Enflurane was made up as a saturated stock solution and diluted in the superfusate; concentrations were measured using gas chromatography. RESULTS: Excitatory postsynaptic currents and EPSPs recorded from motor neurons by stimulation in the dorsal horn were mediated by glutamate receptors of both non NMDA and NMDA subtypes. Enflurane at a general anesthetic concentration (one minimum alveolar anesthetic concentration) reversibly depressed EPSCs and EPSPs. Enflurane also depressed glutamate-evoked currents in the presence of tetrodotoxin (300 nm), showing that its actions are postsynaptic. Block of inhibitory gamma-aminobutyric acid A and glycine receptors by bicuculline (20 micrometer) or strychnine (2 micrometer) or both did not significantly reduce the effects of enflurane on glutamate-evoked currents. Enflurane also depressed glutamate-evoked currents if the inhibitory receptors were blocked and if either D,L-2-amino-5-phosphonopentanoic acid (50 micrometer) or 6-cyano-7 nitroquinoxaline-2,3-dione disodium (10 micrometer) was applied to block NMDA or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-kainate receptors respectively. CONCLUSIONS: Enflurane exerts direct depressant effects on both alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and NMDA glutamate currents in motor neurons. Enhancement of gamma-aminobutyric acid A and glycine inhibition is not needed for this effect. Direct depression of glutamatergic excitatory transmission by a postsynaptic action on motor neurons thus may contribute to general anesthesia as defined by immobility in response to a noxious stimulus. PMID- 11020765 TI - The effect of altered cerebral blood flow on the cerebral kinetics of thiopental and propofol in sheep. AB - BACKGROUND: Thiopental and propofol are highly lipid-soluble, and their entry into the brain often is assumed to be limited by cerebral blood flow rather than by a diffusion barrier. However, there is little direct experimental evidence for this assumption. METHODS: The cerebral kinetics of thiopental and propofol were examined over a range of cerebral blood flows using five and six chronically instrumented sheep, respectively. Using anesthesia (2.0% halothane), three steady state levels of cerebral blood flow (low, medium, and high) were achieved in random order by altering arterial carbon dioxide tension. For each flow state, 250 mg thiopental or 100 mg propofol was infused intravenously over 2 min. To quantify cerebral kinetics, arterial and sagittal sinus blood was sampled rapidly for 20 min from the start of the infusion, and 1.5 h was allowed between consecutive infusions. Various models of cerebral kinetics were examined for their ability to account for the data. RESULTS: The mean baseline cerebral blood flows for the "high" flow state were over threefold greater than those for the low. For the high-flow state the normalized arteriovenous concentration difference across the brain was smaller than for the low-flow state, for both drugs. The data were better described by a model with partial membrane limitation than those with only flow limitation or dispersion. CONCLUSIONS: The cerebral kinetics of thiopental and propofol after bolus injection were dependent on cerebral blood flow, despite partial diffusion limitation. Higher flows produce higher peak cerebral concentrations. PMID- 11020766 TI - Effects of gaseous anesthetics nitrous oxide and xenon on ligand-gated ion channels. Comparison with isoflurane and ethanol. AB - BACKGROUND: Ligand-gated ion channels are considered to be potential general anesthetic targets. Although most general anesthetics potentiate the function of gamma-aminobutyric acid receptor type A (GABAA), the gaseous anesthetics nitrous oxide and xenon are reported to have little effect on GABAA receptors but inhibit N-methyl-d-aspartate (NMDA) receptors. To define the spectrum of effects of nitrous oxide and xenon on receptors thought to be important in anesthesia, the authors tested these anesthetics on a variety of recombinant brain receptors. METHODS: The glycine, GABAA, GABA receptor type C (GABAC), NMDA, alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionic acid (AMPA), kainate, 5-hydroxytryptamine3 (5-HT3), and nicotinic acetylcholine (nACh) receptors were expressed in Xenopus oocytes and effects of nitrous oxide and xenon, and as equipotent concentrations of isoflurane and ethanol, were studied using the two-electrode voltage clamp. RESULTS: Nitrous oxide (0.58 atmosphere [atm]) and xenon (0.46 atm) exhibited similar effects on various receptors. Glycine and GABAA receptors were potentiated by gaseous anesthetics much less than by isoflurane, whereas nitrous oxide inhibited GABAC receptors. Glutamate receptors were inhibited by gaseous anesthetics more markedly than by isoflurane, but less than by ethanol. NMDA receptors were the most sensitive among glutamate receptors and were inhibited by nitrous oxide by 31%. 5-HT3 receptors were slightly inhibited by nitrous oxide. The nACh receptors were inhibited by gaseous and volatile anesthetics, but ethanol potentiated them. The sensitivity was different between alpha4beta2 and alpha4beta4 nACh receptors; alpha4beta2 receptors were inhibited by nitrous oxide by 39%, whereas alpha4beta4 receptors were inhibited by 7%. The inhibition of NMDA and nACh receptors by nitrous oxide was noncompetitive and was slightly different depending on membrane potentials for NMDA receptors, but not for nACh receptors. CONCLUSIONS: Nitrous oxide and xenon displayed a similar spectrum of receptor actions, but this spectrum is distinct from that of isoflurane or ethanol. These results suggest that NMDA receptors and nACh receptors composed of beta2 subunits are likely targets for nitrous oxide and xenon. PMID- 11020767 TI - Periischemic cerebral blood flow (CBF) does not explain beneficial effects of isoflurane on outcome from near-complete forebrain ischemia in rats. AB - BACKGROUND: Isoflurane improves outcome from near-complete forebrain ischemia in rats compared with fentanyl-nitrous oxide (N2O). Sympathetic ganglionic blockade with trimethaphan abolishes this beneficial effect. To evaluate whether anesthesia-related differences in cerebral blood flow (CBF) may explain these findings, this study compared regional CBF before, during, and after near complete forebrain ischemia in rats anesthetized with either isoflurane (with and without trimethaphan) or fentanyl-nitrous oxide. METHODS: Fasted, normothermic isoflurane anesthetized Sprague-Dawley rats were prepared for near-complete forebrain ischemia (10 min of bilateral carotid occlusion and mean arterial pressure = 30 mmHg). After surgery, rats were anesthetized with either 1.4% isoflurane (with or without 2.5 mg of trimethaphan intravenously at onset of ischemia) or fentanyl-nitrous oxide (25 microgram. kg-1. h-1. 70% N2O-1). Regional CBF was determined (14C-iodoantipyrine autoradiography) before ischemia, 8 min after onset of ischemia, and 30 min after onset of reperfusion. RESULTS: Regional CBF did not differ significantly among groups at any measurement interval. Ischemia caused a marked flow reduction to 5% or less of baseline (P < 0.001) in selectively vulnerable regions, such as the cortex, caudoputamen and hippocampus, whereas flow in the brain stem and cerebellum was preserved. Reperfusion at 30 min was associated with partial restoration of flow to 35-50% of baseline values in ischemic structures. CONCLUSIONS: The results indicate that improved histologic-behavioral outcome provided by isoflurane anesthesia cannot be explained by differential vasodilative effects of the anesthetic states before, during, or after severe forebrain ischemia. This study also shows severe postischemic delayed hypoperfusion that was not affected by choice of anesthetic or the presence of trimethaphan. Mechanisms other than effects on periischemic CBF must be responsible for beneficial effects of isoflurane in this model. PMID- 11020768 TI - Statistical method for predicting when patients should be ready on the day of surgery. AB - BACKGROUND: Previously, mathematical theory was developed for determining when a patient should be ready for surgery on the day of surgery. To apply this theory, a method is needed to predict the earliest start time of the case. METHODS: The authors calculated a time estimate such that the probability is 0.05 that the preceding case in the patient's operating room (OR) will be finished before the patient is ready for surgery. This implies there will be a 5% risk of OR personnel being idle and waiting for the patient. This 0. 05 value was chosen by considering the relative cost valuation of an average patient's time to that of an average surgical team based on national salary data. Case duration data from a surgical services information system were used to test different statistical methods to estimate earliest start times. RESULTS: Simulations found that 0.05 prediction bounds, calculated assuming case durations followed log-normal distributions, achieved actual risks for the OR staff to wait for patients of 0.050 to 0.053 (SEM = 0.001). Nonparametric prediction bounds performed no better than the parametric method. Having patients ready a fixed number of hours before the scheduled starts of their operations is not reliable. If the preceding case in an OR had been underway for 0.5 to 1.5 h, the parametric 0.05 prediction bounds for the time remaining achieved actual risks for OR staff waiting of 0.055 to 0.058 (SEM = 0.001). CONCLUSION: The earliest start time of a case can be estimated using the 0.05 prediction bound for the duration of the preceding case. The authors show 0.05 prediction bounds can be estimated accurately assuming that case durations follow log-normal distributions. PMID- 11020769 TI - Systematic overview of the evidence supporting the use of cerebrospinal fluid drainage in thoracoabdominal aneurysm surgery for prevention of paraplegia. PMID- 11020770 TI - Chronic pain as an outcome of surgery. A review of predictive factors. PMID- 11020771 TI - Survey of residency training in preoperative evaluation. PMID- 11020772 TI - Preemptive analgesia. PMID- 11020773 TI - Systemic anaphylaxis associated with intramammary isosulfan blue injection used for sentinel node detection under general anesthesia. PMID- 11020774 TI - Factious desaturation due to isosulfan dye injection. PMID- 11020775 TI - Bilateral intraoperative atelectasis in a child with latex allergy. PMID- 11020776 TI - Hyperbaric oxygen reverses organ dysfunction in severe anemia. PMID- 11020777 TI - Can epidural fentanyl induce selective spinal hyperalgesia? PMID- 11020778 TI - How to open the Lung? The unsolved question. PMID- 11020779 TI - Pulmonary aspiration of a milk-cream mixture. PMID- 11020780 TI - Pulmonary aspiration of milk and cream: An avoidable complication. PMID- 11020781 TI - Venous air embolism in craniosynostosis surgery: what do we want to detect? PMID- 11020782 TI - In vitro model for cardiac function under Xenon anesthesia. PMID- 11020783 TI - Combination of two standard pneumatic calf compression devices to fit the morbidly obese. PMID- 11020784 TI - Anesthetic implications of a new implantable device: the loop recorder system. PMID- 11020785 TI - Henry Isaiah Dorr was the first person to hold the title professor of anaesthesia. PMID- 11020786 TI - A modest proposal. PMID- 11020787 TI - Life history of Aptesis nigrocincta (Hymenoptera: Ichneumonidae) a cocoon parasitoid of the apple sawfly, Hoplocampa testudinea (Hymenoptera: Tenthredinidae). AB - Aptesis nigrocincta: Gravenhorst is a bivoltine ectoparasitoid of apple sawfly cocoons, hosts that must be found and parasitized by females at a depth of 10-25 cm in the soil. Females are significantly smaller than males and nearly wingless. After encountering a host, females needed 29.3 min at 20 degrees C and 19. 9 min at 25 degrees C to deposit an egg on the host. Development from egg to adult took 39.6 days for females and 38.0 days for males at 20 degrees C. This small difference was significant. At 20 degrees C, the longevity of females that had no opportunity to oviposit was on average 72.5 days, significantly higher than male longevity (50.6 days). The longevity of females given access to hosts throughout their lifetime averaged 58.6 days. Females were able to mate immediately after emergence and copulation lasted on average 21.7 s. After a pre-oviposition period averaging 5.8 days, females laid 20.2 eggs during their lifetime, thus less than one egg per day. Neither the fecundity nor longevity of individual females was correlated with body size. If females were deprived of food, longevity as well as lifetime fecundity were drastically reduced. Field studies were carried out in one organically managed apple orchard in Switzerland. Aptesis nigrocincta showed parasitism rates ranging from 12.1 to 39.7 % within single parasitoid generations, thereby constituting the most important mortality factor of apple sawfly cocoons. PMID- 11020788 TI - The biology of Meteorus gyrator (Hymenoptera: Braconidae), a solitary endoparasitoid of the tomato moth, Lacanobia oleracea (Lepidoptera: Noctuidae). AB - There is a need to identify potential biological control agents for use against lepidopterous pests in greenhouses. The solitary endoparasitoid Meteorus gyrator (Thunberg) attacks a range of macrolepidopterous larvae, including those of some important horticultural pest species. Laboratory trials designed to investigate the biology of M. gyrator on larvae of the tomato moth, Lacanobia oleracea Linnaeus, reveal that this parasitoid is capable of parasitizing all larval stages of its host, third instars being parasitized most frequently. Each female parasitoid lives for up to 40 days (at 25 degrees C), ovipositing into an average of 78 hosts. Preadult development is rapid ( approximately 2 weeks), and the sex ratio of offspring is 1:1. Parasitism by M. gyrator suppresses the growth of both early and late host instars, and there is a concomitant reduction in the amount of food consumed (overall feeding reduction over a 12 day period is 68%). Our results indicate that inoculative releases of M. gyrator could provide effective biological control of L. oleracea and other noctuid pests of greenhouses. PMID- 11020789 TI - Population biology of the potato tuber moth, Phthorimaea operculella (Lepidoptera: Gelechiidae), in two potato cropping systems in Israel. AB - The life cycle, within-field distribution, crop damage and impact of natural enemies of the potato tuber moth, Phthorimaea operculella (Zeller) were investigated in two potato cropping systems. The two systems differed in soil type (sandy vs. loess) which in turn affected the choice of cultivars, irrigation programme, insecticide application method (ground vs. aerial), and planting and harvest times. From mid-April to the end of May, almost twice as many moths were caught in pheromone traps in sandy than in loess fields. Highest infestation of tubers was found before harvest, and infestation was greater in loess than in sandy fields. Larval densities in foliage and tubers were significantly higher at the margins of the fields than in the centre. A significant positive correlation was found between adult catch and larval infestation on foliage in sandy fields but not in loess. Tuber infestation in sand was positively correlated with foliage infestation. No such correlation was detected in loess. Five parasitic wasps emerged from P. operculella larvae collected from commercial fields and volunteer plants: Diadegma pulchripes (Kokujev) and Temelucha decorata, (Gravenhorst) (Ichneumonidae) and Bracon gelechiae Ashmead and two other unidentified Braconidae. The most abundant predators at the field site were Coccinella septempunctata Linnaeus (Coccinellidae), Chrysoperla carnea Stephens (Chrysopidae), Orius albidipennis (Reuter) (Anthocoridae) and four ant species (Formicidae). Parasitism rate reached 40% and predation was estimated at 79%. Results are discussed with regard to the development of an integrated pest management programme for this important pest. PMID- 11020790 TI - Variation in the Bemisia tabaci s. l. species complex (Hemiptera: Aleyrodidae) and its natural enemies leading to successful biological control of Bemisia biotype B in the USA. AB - Parasitoids of the Bemisia tabaci (Gennadius) species complex collected in Spain and Thailand were evaluated as biological control agents of B. tabaci biotype B in cole crops in Texas, USA. Parasitoids were identified by morphological and RAPD-PCR analyses. The most abundant parasitoid from Spain was Eretmocerus mundus Mercet with apparent field parasitism of 39-44%. In Thailand, Encarsia formosa Gahan, E. transvena Timberlake, E. adrianae Lopez-Avila, Eretmocerus sp. 1 and sp. 2 emerged, with apparent field parasitism of 1-65%. Identification and molecular classification of B. tabaci associated with parasitoid collections and in the release site in Texas were accomplished using morphological traits and nucleotide sequence comparison of the mitochondrial cytochrome oxidase I gene (COI) (700-720 bp). Collections of B. tabaci from Thailand grouped separately from B types from Arizona and Florida and the target B type from Texas, USA, a cluster from India, and other New World B. tabaci. The Spanish B. tabaci host of E. mundus which was laboratory and field-tested to achieve biological control of the B type was most closely related to non-B type B. tabaci populations from Spain and Sudan, the latter which formed a second group within the larger clade that also contained the B type cluster. Laboratory tests indicated that E. mundus from Spain parasitized more B. tabaci type B than did Eretmocerus spp. native to Texas and other exotic parasitoids evaluated. Eretmocerus mundus from Spain also successfully parasitized B. tabaci type B when field-released in a 0.94 million ha test area in Texas, and has significantly enhanced control of B. tabaci type B in California, USA. In contrast, parasitoids from Thailand failed to establish in the field in Texas, collectively suggesting a positive correlation between the centres of diversity of compatible parasitoid-host complexes. PMID- 11020791 TI - Population genetics of Glossina morsitans submorsitans (Diptera: Glossinidae). AB - Breeding structure of Glossina morsitans submorsitans Newstead was evaluated by using genetic markers in mitochondrial DNA where diversity was scored at two loci in five natural populations from The Gambia and two populations in Ethiopia (form ugandensis Vanderplank), countries separated by c. 5450 km. Twenty six haplotype combinations were found, of which 17 were shared among two or more populations. Nine haplotypes were found in The Gambia and 23 haplotypes in Ethiopia. There were 12 unique haplotypes. Only six haplotypes were shared between the two countries. Populations in The Gambia (he = 0.26 +/- 0.04) showed less than a third of the diversity of populations in Ethiopia (he = 0.84 +/- 0.03). This suggests recovery from an earlier reduction in population. In a nested analysis of molecular variance of haplotype frequencies, 65% of the variance was due to differences within populations, 34% to differences between populations grouped by country, and only 1% was due to differences among populations within countries. In terms of gene flow, the fixation index FST = 0.35, which leads to an estimate by Wright's island model of less than one reproducing migrant per generation exchanged between the eastern and western submorsitans populations. Nei's genetic similarity measure showed a deep division between Gambian and Ethiopian populations. PMID- 11020792 TI - Modelling development time of Lipaphis erysimi (Hemiptera: Aphididae) at constant and variable temperatures. AB - The development period from birth to adult of virginoparae of the turnip aphid, Lipaphis erysimi (Kaltenbach), at 14 constant, 15 alternating and 15 natural temperature regimes were modelled to determine mathematical functions for simulating aphid development under a wide range of natural conditions. The day degree model, the logistic equation, and the Wang model were used to describe the relationships between temperature and development rate at constant and alternating temperatures. The three models were then used with a Weibull function describing the distribution of development times, to simulate the development of individuals of cohorts at natural temperature regimes. Comparison of the observed with simulated distributions of adult emergence indicates that all three models can simulate the development of L. erysimi equally well when temperature does not go below 6 degrees C (the notional low temperature threshold of the day-degree model) or above 30 degrees C. When accumulation of temperatures below 6 degrees C becomes substantial, only the logistic curve offers accurate simulations; the other two models give falsely longer durations of development. When accumulation of temperatures above 30 degrees C becomes substantial, the logistic curve and the Wang model offer more accurate simulations than the day-degree model, which tends to produce shorter durations of development. Further analysis of the data reveals that development rate of this aphid at a given unfavourable high temperature may vary with time. Methods for accurately simulating the development time of L. erysimi in the field are suggested. The significance of modelling insect development at low and high temperatures by non-linear models is discussed. PMID- 11020793 TI - Advantageous attributes of larval whitefringed weevil, Naupactus leucoloma (Coleoptera: Curculionidae) for bioassaying soil fumigants, and responses to pure and plant-derived isothiocyanates. AB - First instars of the soil-inhabiting whitefringed weevil, Naupactus leucoloma (Boheman), are a particularly good bioassay model for assessing volatile soil fumigants and biofumigants. Eggs are readily obtained and can be stored for long periods with larvae hatched on demand and the first instar is non-feeding, surviving without food or shelter. Longevity varies with temperature, but readily accommodates the period required to conduct bioassays without appreciable mortality of untreated controls. In vitro bioassays of pure methyl isothiocyanate, the active ingredient from metham sodium soil fumigant, and the less volatile 2-phenylethyl isothiocyanate, sensitively detected differences in toxicity and effects of temperature. Bioassay of volatiles emitted from hydrolysed tissue of various isothiocyanate-producing Brassica plants revealed widely varying toxicity effects, indicating that bioassays with N. leucoloma are a sensitive and relevant indicator of the potential of different plants for biofumigation of soil-borne pest organisms. PMID- 11020794 TI - Temperature tolerance of the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae) in tropical and temperate regions of Asia. AB - Temperature tolerance was investigated in nine populations of Plutella xylostella Linnaeus from tropical and temperate regions of Asia. At all rearing temperatures between 15 and 35 degrees C, no clear differences were observed in female egg production or larval development between tropical and temperate populations. Thus, tropical populations did not show a high-temperature tolerance superior to that of the temperate populations. In all populations, the net reproductive rate (number of new females born per female) largely depended on the number of eggs laid per female, and egg production significantly decreased with increasing temperature (P < 0.001). Larval developmental rate also showed a significant positive correlation with temperature (P < 0.001). Per cent hatch of eggs and larval survival did not show a significant correlation with temperature: hatching was constant between 15 and 32.5 degrees C, but considerably lower at 35 degrees C. Larval survival was similar between 15 and 30 degrees C, appreciably lower at 32.5 degrees C and declined to 0% at 35 degrees C. Based on these results, environmental conditions under which P. xylostella can maintain a high population density throughout the year in tropical and subtropical regions are discussed. PMID- 11020795 TI - Diapause initiation and incidence in the millet stem borer, Coniesta ignefusalis (Lepidoptera: Pyralidae): the role of the host plant. AB - The role of the host plant in the development of larval diapause in the millet stem borer, Coniesta ignefusalis (Hampson) was investigated in northern Ghana in 1996 and 1997. Surveys conducted in farmers' fields in the Guinea and Sudan savannah revealed that of all the upland cereals grown, the insect survived the dry season only in stalks and stubble of pearl millet, Pennisetum glaucum and late sorghum, Sorghum bicolor. This observation was confirmed by results from field trials conducted at the Manga Research Station. In these studies, C. ignefusalis larvae entered diapause only in late millet and late sorghum, with a higher incidence in the former. The insect neither attacked nor entered diapause in maize planted during the same period as the other crops. Results from controlled experiments showed that diapause incidence in the preferred host, millet, was higher in older than in younger plants, suggesting that host plant maturation is a key factor influencing the development of larval diapause in C. ignefusalis. PMID- 11020796 TI - Regulated expression of an MHC class II gene from a promoter-inducible retrovirus. AB - Specific immune tolerance to fully allogeneic kidney grafts can be achieved in a miniature swine transplantation model by retrovirus-mediated transfer of allogeneic MHC class II genes into bone marrow cells (BMCs) of recipient animals. Graft survival correlated with transient expression of the somatic transgene (Tg) in the induction phase of tolerance. With the aim of investigating the effects of timing and threshold levels of Tg expression on induction of hyporesponsiveness to the grafted tissues, two recombinant retrovirus constructs containing the tetracycline binary regulatory system were used to achieve conditional expression of either the green fluorescent protein (tetGFP) as a control, or the porcine MHC class II DRbeta chain (tetDRB). Effective downregulation of GFP gene transcription was demonstrated in transduced murine fibroblasts after doxycycline treatment, leading to a > 90% reduction of GFP fluorescence. Similar diminution of the DRB gene transcription was achieved in transduced pig endothelial cells (ECs). Drug-dependent downregulation of DRBc gene expression in SLAd pig ECs coincided with complete inhibition of allogeneic activation of anti-class IIc primed SLAd T cells. These in vitro results suggest that the binary tetracycline retrovirus system may also be adequate to regulate MHC class II Tg expression in vivo. PMID- 11020797 TI - Molecular adaptors for vascular-targeted adenoviral gene delivery. AB - Gene therapy would be considerably more effective if vectors could be targeted to specific organs or tissues after systemic administration. We previously developed an in vivo selection system to isolate organ- and tumor-homing peptides from phage display peptide libraries. The peptides isolated by this approach bind to receptors expressed in vascular endothelia. We describe here the development of molecular adaptors to target adenoviral gene therapy vectors to selective vascular "addresses." The adaptor design consists of an organhoming peptide conjugated to an adenovirus-binding moiety. We isolated and characterized several monoclonal antibodies that bind to adenovirus type 5 (Ad5). Two of the antibodies neutralized Ad5 infection. We linked the Fab fragments of one of these antibodies to a synthetic lung-homing peptide (CGFECVRQCPERC or GFE-1 peptide) and tested the ability of the resulting bispecific conjugate to retarget Ad5. Cells that express the receptor for the GFE-1 peptide and are resistant to Ad5 infection were sensitized to recombinant Ad5 vectors in the presence of the Fab-GFE adaptor. Our findings indicate that selective gene therapy delivery may be developed on the basis of our vascular targeting technology. PMID- 11020798 TI - Antiangiogenic gene therapy for cancer via systemic administration of adenoviral vectors expressing secretable endostatin. AB - A growing number of antiangiogenesis strategies have been investigated for the treatment of cancer and other angiogenesis-dependent diseases. One of the most promising strategies is to systemically administer one or more antiangiogenic proteins frequently enough to achieve a sufficient long-term steady state level of the protein(s) to achieve the maximum beneficial effect. However, the utility of this strategy is limited because of many technical difficulties, including obtaining both the quantity and quality of the protein(s) necessary for optimal therapeutic benefit. To overcome these difficulties, we hypothesized that a single administration of a replication-defective adenoviral vector expressing a secretable antiangiogenic protein could achieve an optimal long-term systemic concentration. We constructed a recombinant adenoviral vector, Av3mEndo, which encodes a secretable form of murine endostatin. We demonstrated secretion of endostatin from several cell lines transduced with Av3mEndo. Partially purified endostatin secreted from Av3mEndo-transduced mammalian cells was shown to potently inhibit endothelial cell migration in vitro. A single intravenous administration of Av3mEndo in mice was shown to result in (1) prolonged and elevated levels of circulating endostatin, (2) partial inhibition of VEGF-induced angiogenesis in a VEGF implant angiogenesis model, and (3) prolonged survival and in 25% of mice the complete prevention of tumor growth in a prophylactic human colon/liver metastasis xenograft murine model. These results support our contention that adenoviral vector-mediated expression of an antiangiogenic protein(s) represents an attractive therapeutic approach to cancer and other angiogenesis-dependent diseases. PMID- 11020799 TI - Inhibitory effect of cystic fibrosis sputum on adenovirus-mediated gene transfer in cultured epithelial cells. AB - Effective gene transfer to the airway epithelial cells of individuals with cystic fibrosis (CF) requires gene therapy vectors to effectively penetrate the mucous lining of the airways of these patients. In this study, we examined the effects of the aqueous sol fraction of sputum recovered from CF patients (CF sol) on adenovirus (Ad)-mediated gene transfer to cultured epithelial cells. Sputum collected from patients with CF was separated into aqueous sol and gel fractions by ultracentrifugation and the sol fraction from different individuals was pooled. To determine if CF sol affects Ad-mediated transfection, Fisher rat thyroid (FRT) epithelial cells or normal human bronchial epithelial (NHBE) cells were infected with an Ad encoding beta-galactosidase (Ad2/betagal-2) in the presence or absence of the pooled CF sol. Transfection efficiency was determined by measuring beta-Gal activity. CF sol significantly inhibited Ad2-mediated gene transfer in a dose-dependent manner when the vector was incubated with CF sol prior to exposure to the cells. In contrast, preincubation of the cells with the sol was without effect. The inhibition of Ad-mediated gene transfer by CF sol was not related to its low pH, was abrogated by preadsorption with an Ad2 serotype vector, and was neutralized by heat treatment, but was not affected by treatment with protease inhibitors. Analysis of CF sol fractions from seven different individuals with CF showed inhibition of Ad-mediated gene transfer in four of the seven samples tested and, further, the inhibitory effect was correlated with the presence of Ad-specific antibodies. We conclude that preexisting adenovirus specific antibodies present in some of the patient samples were the predominant factor inhibiting Ad-mediated gene transfer. PMID- 11020801 TI - Potent inhibition of human immunodeficiency virus type 1 replication by conditionally replicating human immunodeficiency virus-based lentiviral vectors expressing envelope antisense mRNA. AB - We describe an HIV-based lentiviral vector that expresses a 1-kb antisense mRNA directed against the HIV-1 mRNAs containing env sequences. The expression of antisense env mRNAs (envAS) does not inhibit the synthesis of p24 expressed from the HIV-1 helper plasmid used to package the vector, as this helper has a deletion in the env gene. This allows the production of high-titer VSV-G pseudotyped lentiviral particles. In challenge experiments using unselected populations of SupT1 cells transduced with this vector, a complete inhibition of HIV-1 replication was observed for long periods of in vitro culture, even at high HIV-1 infectious doses. The potent inhibition of HIV-1 replication by this vector correlated with a low occurrence of mobilization of the vector to previously untransduced cells. The infectivity of the wild-type HIV-1 that escapes inhibition was highly inhibited, suggesting that the vector is providing HIV-1 inhibition of replication not only due to its antisense effect but also by competing for encapsidation and mobilization to noninfected cells. PMID- 11020800 TI - A novel, conditionally replicative adenovirus for the treatment of breast cancer that allows controlled replication of E1a-deleted adenoviral vectors. AB - The efficiency of gene therapy strategies against cancer is limited by the poor distribution of the vectors in the malignant tissues. To solve this problem, a new generation of tumor-specific, conditionally replicative adenoviruses is being developed. To direct the replication of the virus to breast cancer, we have considered one characteristic present in a great proportion of these cancers, which is the expression of estrogen receptors (ERs). On the basis of the wild type adenovirus type 5, we have constructed a conditionally replicative adenovirus (Ad5ERE2) in which the E1a and E4 promoters have been replaced by a portion of the pS2 promoter containing two estrogen-responsive elements (EREs). This promoter induces transcriptional activation of the E1a and E4 units in response to estrogens in cells that express the ERs. Ad5ERE2 is able to kill ER(+) human breast cancer cell lines as efficiently as the wild-type virus, but has decreased capacity to affect ER(-) cells. By complementation of the E1a protein in trans, Ad5ERE2 allows restricted replication of a conventional E1a deleted adenoviral vector. When a virus expressing the proapoptotic gene Bc1-xs (Clarke et al., Proc. Natl. Acad. Sci. U.S.A. 1995;92:11024-11028) is used in combination with Ad5ERE2, the ability of both viruses to induce cell death is dramatically increased, and the effect can be modulated by addition of the antiestrogen tamoxifen. PMID- 11020802 TI - Addition of the human interferon beta scaffold attachment region to retroviral vector backbones increases the level of in vivo transgene expression among progeny of engrafted human hematopoietic stem cells. AB - Absence of durable high-level expression of transgenes from Moloney murine leukemia (Mo-MuLV) retroviral vectors has been a hurdle in bringing effective gene therapy to the clinic. In this study we have analyzed transgene expression among the progeny of mobilized hematopoietic stem cells (HSCs), comparing Mo-MuLV and mouse stem cell virus (MSCV) vectors, with or without addition of a scaffold attachment region (SAR) from the human interferon beta gene. Retroviral (RV) vector supernatant quality was assessed by comparing NGFR transgene expression by HEL cells, and transgene delivery and expression by CD34(+) cells 72 hr after transduction, using real-time PCR and FACS analysis. This is the first description of the effect of SAR within both Mo-MuLV and MSCV vector backbones on long-term RV transgene expression among in vivo HSC progeny in HSC repopulation assays (SCID-hu bone and NOD/SCID). After transduction of mobilized CD34(+) cells with MSCV-SAR vector, transgene expression was observed among a mean of 10% of donor HSC progeny in the SCID-hu bone (range, 0.6-43%). The predominant effect of SAR was to increase the mean fluorescence intensity (MFI) of transgene expression among HSC progeny in both in vivo bone repopulation models (three- to fourfold), and after long-term stromal cultures (twofold). PMID- 11020803 TI - In vivo competitive studies between normal and common gamma chain-defective bone marrow cells: implications for gene therapy. AB - Corrective gene transfer into hematopoietic stem cells (HSCs) is being investigated as therapy for X-linked severe combined immunodeficiency (XSCID) and it is hoped that selective advantage of gene-corrected HSCs will help in achieving full immune reconstitution after treatment. Lines of evidence from the results of allogeneic bone marrow transplantation in patients with XSCID support this hypothesis that, however, has not been rigorously tested in an experimental system. We studied the competition kinetics between normal and XSCID bone marrow (BM) cells using a murine bone marrow transplantation (BMT) model. For easy chimerism determination, we used genetic marking with retrovirus-mediated expression of the enhanced green fluorescent protein (EGFP). We found that XSCID BM cells were able to compete with normal BM cells for engraftment of myeloid lineages in a dose-dependent manner, whereas we observed selective repopulation of T, B, and NK cells deriving from normal BM cells. This was true despite the evidence of competitive engraftment of XSCID lineage marker-negative/c-Kit positive (Lin-/c-Kit+) cells in the bone marrow of treated animals. From these results we extrapolate that genetic correction of XSCID HSCs will result in selective advantage of gene-corrected lymphoid lineages with consequent restoration of lymphocyte populations and high probability of clinical benefit. PMID- 11020804 TI - Regulatory issues: future meetings of the NIH recombinant DNA advisory committee (RAC) and gene therapy policy conferences (GTPC) PMID- 11020805 TI - Biomedical information technology: medicine and health care in the digital future. AB - Advancements in medicine and health care are being significantly influenced by the exploding information technology developments. The IEEE Transactions on Information Technology in Biomedicine will address the applications and the infrastructure innovations that would harness biomedical and health care programs in the 21st century. PMID- 11020806 TI - What can you publish in T-ITB? AB - IEEE Transactions on Information Technology in Biomedicine will focus on applications reflecting global information technology advances in medicine and biology and will further address the implementation and management of the broad spectrum of health care innovations arising from these developments worldwide. PMID- 11020807 TI - Programmable ultrasound imaging using multimedia technologies: a next-generation ultrasound machine. AB - High computational and throughput requirements in modern ultrasound machines have restricted their internal design to algorithm-specific hardware with limited programmability. We have architected a programmable ultrasound processing system, Programmable Ultrasound Image Processor (PUIP), to facilitate engineering and clinical ultrasound innovations. Multiple high-performance multimedia processors were used to provide a computing power of 4 billion operations per second. Flexibility was achieved by making our system programmable and multimodal, e.g., B-mode, color flow, cine and Doppler data can be processed. We have successfully designed and implemented the PUIP to fit within an ultrasound machine. It provides a platform for rapid testing of new concepts in ultrasound processing and enables software upgrades for future technologies. Current and future clinical applications include extended fields of view, quantitative measurements, three-dimensional ultrasound reconstruction and visualization, adaptive persistence, speckle reduction, edge enhancement, image segmentation, and motion analysis. The PUIP is a significant step in the evolution of ultrasound machines toward more flexible and generalized systems bridging the gap between many innovative ideas and their clinical use in ultrasound machines. PMID- 11020808 TI - A direct multi-volume rendering method aiming at comparisons of 3-D images and models. AB - We present a new method for direct volume rendering of multiple three-dimensional (3-D) functions using a density emitter model. This work aims at obtaining visual assessment of the results of a 3-D image registration algorithm which operates on anisotropic and non segmented medical data. We first discuss the fundamentals associated with direct, simultaneous rendering of such datasets. Then, we recall the fuzzy classification and fuzzy surface rendering theory within the density emitter model terminology, and propose an extension of standard direct volume rendering that can handle the rendering of two or more 3-D functions; this consists of the definition of merging rules that are applied on emitter clouds. The included rendering applications are related on one hand, to volume-to-volume registration, and on the other hand, to surface-to-volume registration: the first case is concerned with global elastic registration of CT data, and the second one presents fitting of an implicit surface over a CT data subset. In these two medical imaging application cases, our rendering scheme offers a comprehensive appreciation of the relative position of structural information. PMID- 11020809 TI - A nurse rostering system using constraint programming and redundant modeling. AB - This paper describes the design and implementation of a constraint-based nurse rostering system using a redundant modeling approach. Nurse rostering is defined as the process of generating timetables for specifying the work shifts of nurses over a given period of time. This process is difficult because the human roster planner has to ensure that every rostering decision made complies with a mixture of hard hospital rules and soft nurse preference rules. Moreover, some nurse shift pre-assignments often break the regularity of wanted (or unwanted) shifts and reduce the choices for other unfilled slots. Soft constraints amount to disjunction, which can be modeled as choices in the search space. This approach, although straightforward, incurs overhead in the search of solution. To reduce search time, we propose redundant modeling, an effective way to increase constraint propagation through cooperations among different models for the same problem. Our problem domain involves around 25 to 28 nurses and 11 shift types. Experiments and pilot testing of the system confirm the effectiveness and efficiency of our method. PMID- 11020810 TI - Compression guidelines for diagnostic telepathology. AB - As the healthcare community has begun to rely increasingly upon digital technologies for acquisition, storage, and transmission of pictorial data, image compression has become an indispensable tool. We have investigated the feasibility of lossy compression in a well-defined task domain, the clinical assessment of digitized images of chromatic microscopic pathology specimens. The effect of compression was measured under two distinct perceptual criteria, just noticeable difference (j.n.d.) and largest tolerable distortion (l.t.d.), differing in the involvement required from subjects, who were experts in pathology. For standard JPEG compressed images it was found that when the experiment is performed under the l.t.d. criterion, a significantly larger compression ratio is reported as satisfactory. It is concluded that lossy compression holds promise for diagnostic telepathology. PMID- 11020811 TI - Virtual reality in medicine-computer graphics and interaction techniques. AB - This paper describes several new visualization and interaction techniques that enable the use of virtual environments for routine medical purposes. A new volume rendering method supports shaded and transparent visualization of medical image sequences in real-time with an interactive threshold definition. Based on these rendering algorithms two complementary segmentation approaches offer an intuitive assistance for a wide range of requirements in diagnosis and therapy planning. In addition, a hierarchical data representation for geometric surface descriptions guarantees an optimal use of available hardware resources and prevents inaccurate visualization. The combination of the presented techniques empowers the improved human-machine interface of virtual reality to support every interactive task in medical three-dimensional (3-D) image processing, from visualization of unsegmented data volumes up to the simulation of surgical procedures. PMID- 11020812 TI - An architecture for naval telemedicine. AB - Navy fleets have a defined overall objective for mission readiness impacted by the health of personnel aboard the ships. Medical treatment facilities on the ships determines the degree of mission readiness. This paper describes the concepts and technologies necessary to establish a Naval telemedicine system, which can drastically improve health care delivery. It consists of various combinations of the following components: Fleet Naval Medical Consultation and Diagnostic Centers, Shipboard Naval Medical Consultation and Diagnostic Centers (hospital ship or combatant ships with medical specialists on board), and Remote Medical Referring Centers such as a ship, a small Naval station annex, or a field hospital. This Naval telemedicine architecture delivers clinical medicine and continuing medical education (CME) by means of computers, video-conferencing systems, or telephony to enhance the quality of care through improved access to research, medical and nonmedical imaging, remote consultations, patient clinical data, and multimedia medical education programs. It integrates the informatics infrastructure and provides a medical telepresence among participants. PMID- 11020813 TI - Information analysis and validation of intelligent monitoring systems in intensive care units. AB - Validation of intelligent systems is an important task to perform. Typically the results of the validation analysis are used to verify whether or not the system satisfies the initial design requirements, and to acquire new knowledge and/or refine the knowledge already acquired. In practice, the validation of intelligent systems usually requires the application of several different techniques (e.g., retrospective, prospective, quantitative). In this work the authors present the methodology devised to validate PATRICIA: an intelligent monitoring system designed to advise clinicians on the management of patients dependent on mechanical ventilation. The application of this methodology requires that appropriate validation paradigms are selected, depending on both the application domain and the characteristics of the intelligent system. The article also presents and discusses validation results. PMID- 11020814 TI - Applying object-oriented technologies in modeling and querying temporally oriented clinical databases dealing with temporal granularity and indeterminacy. AB - The need of managing temporal information given at different levels of granularity or with indeterminacy is common to many application areas. Among them, we focus on clinical data management. Different time granularities and indeterminacy are also needed in querying temporal databases. In this paper, we describe GCH-OSQL (Granular Clinical History-Object Structured Query Language), an object-oriented temporally-oriented extension of SQL. GCH-OSQL is based on an object-oriented temporal data model, GCH-OODM. GCH-OODM allows storage of clinical information at different and mixed granularities or with temporal indeterminacy. GCH-OSQL deals with the valid time of clinical information. The temporal extension of the SELECT construct includes the addition of the TIME SLICE and MOVING WINDOW clauses, and the capability to reference the temporal dimension of objects in the WHERE and SELECT clauses. Using object-oriented technologies, a system prototype for GCH-OSQL and GCH-OODM has been implemented and applied to data management of follow-up patients after coronary angioplasty intervention. PMID- 11020815 TI - Computer-aided detection of breast cancer nuclei. AB - A computer-aided detection system for tissue cell nuclei in histological sections is introduced and validated as part of the Biopsy Analysis Support System (BASS). Cell nuclei are selectively stained with monoclonal antibodies, such as the anti estrogen receptor antibodies, which are widely applied as part of assessing patient prognosis in breast cancer. The detection system uses a receptive field filter to enhance negatively and positively stained cell nuclei and a squashing function to label each pixel value as belonging to the background or a nucleus. In this study, the detection system assessed all biopsies in an automated fashion. Detection and classification of individual nuclei as well as biopsy grading performance was shown to be promising as compared to that of two experts. Sensitivity and positive predictive value were measured to be 83% and 67.4%, respectively. One major advantage of BASS stems from the fact that the system simulates the assessment procedures routinely employed by human experts; thus it can be used as an additional independent expert. Moreover, the system allows the efficient accumulation of data from large numbers of nuclei in a short time span. Therefore, the potential for accurate quantitative assessments is increased and a platform for more standardized evaluations is provided. PMID- 11020816 TI - Segmentation algorithms for detecting microcalcifications in mammograms. AB - The presence of microcalcification clusters in mammograms contributes evidence for the diagnosis of early stages of breast cancer. In many cases, microcalcifications are subtle and their detection can benefit from an automated system serving as a diagnostic aid. The potential contribution of such a system may become more significant as the number of mammograms screened increases to levels that challenge the capacity of radiology clinics. Many techniques for detecting microcalcifications start with a segmentation algorithm that indicates all candidate structures for the subsequent phases. Most algorithms used to segment microcalcifications have aspects that might raise operational difficulties, such as thresholds or windows that must be selected, or parametric models of the data. We present a new segmentation algorithm and compare it to two other algorithms: the multi-tolerance region growing algorithm that operates without the aspects mentioned above, and the active contour model that has not been applied previously to segment microcalcifications. The new algorithm operates without threshold or window selection, or parametric data models, and it is more than an order of magnitude faster than the other two. PMID- 11020817 TI - Microcontroller-based underwater acoustic ECG telemetry system. AB - This paper presents a microcontroller-based underwater acoustic telemetry system for digital transmission of the electrocardiogram (ECG). The system is designed for the real time, through-water transmission of data representing any parameter, and it was used initially for transmitting in multiplexed format the heart rate, breathing rate and depth of a diver using self-contained underwater breathing apparatus (SCUBA). Here, it is used to monitor cardiovascular reflexes during diving and swimming. The programmable capability of the system provides an effective solution to the problem of transmitting data in the presence of multipath interference. An important feature of the paper is a comparative performance analysis of two encoding methods, Pulse Code Modulation (PCM) and Pulse Position Modulation (PPM). PMID- 11020818 TI - Improvement of sensitivity of breast cancer diagnosis with adaptive neighborhood contrast enhancement of mammograms. AB - Mammograms are difficult to interpret, especially of cancers at their early stages. In this paper, we analyze the effectiveness of our adaptive neighborhood contrast enhancement (ANCE) technique in increasing the sensitivity of breast cancer diagnosis. Seventy-eight screen-film mammograms of 21 difficult cases (14 benign and seven malignant), 222 screen-film mammograms of 28 interval cancer patients and six benign control cases were digitized with a high-resolution of about 4096 x 2048 x 10-bit pixels and then processed with the ANCE method. Unprocessed and processed digitized mammograms as well as the original films were presented to six experienced radiologists for a receiver operating characteristic (ROC) evaluation for the difficult case set and to three reference radiologists for the interval cancer set. The results show that the radiologists' performance with the ANCE-processed images is the best among the three sets of images (original, digitized, and enhanced) in terms of area under the ROC curve and that diagnostic sensitivity is improved by the ANCE algorithm. All of the 19 interval cancer cases not detected with the original films of earlier mammographic examinations were diagnosed as malignant with the corresponding ANCE-processed versions, while only one of the six benign cases initially labeled correctly with the original mammograms was interpreted as malignant after enhancement. McNemar's tests of symmetry indicated that the diagnostic confidence for the interval cancer cases was improved by the ANCE procedure with a high level of statistical significance (p-values of 0.0001-0.005) and with no significant effect on the diagnosis of the benign control cases (p-values of 0.08-0.1). This study demonstrates the potential for improvement of diagnostic performance in early detection of breast cancer with digital image enhancement. PMID- 11020819 TI - Real-time multimodal medical image processing: a dynamic volume-rendering application. AB - In the present research, a system for medical image processing has been proposed, which allows multimodal dynamic three-dimensional (3-D) visualization interactively and in real time. The system has been conceived to support medical specialists in the diagnosis of moving organs, such as the heart during the cardiac cycle, allowing them to compare information on perfusion/contraction match as a basis for diagnosis of important cardiovascular diseases. The 3-D volume-rendering algorithm runs on a SIMD machine because of the great amount of data to be manipulated by always using the same operations. One of the features of the algorithm is the possibility to change, interactively, image processing and visualization parameters at any step, and to perform simple and effective image manipulations. Performance studies have demonstrated a very high global efficiency in practical situations by using typical data-volume dimensions. The system has been tested in the medical environment, by using magnetic resonance (MR) and single-photon emission-computed tomographic (SPECT) images. PMID- 11020820 TI - Design, analysis, and implementation of a telemedicine remote consultation and diagnosis session playback using discrete event system specification. AB - Telemedicine remote consultation and diagnosis (RCD) software is a complex and distributed system. RCD allows physicians to collaborate on radiology or pathology cases from distributed geographic locations. It is very important to simplify design, construction, and maintenance of such a system. Currently, object-oriented design methodology is used to design and develop a software system in a modular fashion. Object-oriented software is made of various objects that work together. From the design of the software system, we get information about object methods and inheritance. We also get information about which objects are contained in a particular object and which objects are used by another object. One important element that the traditional object-oriented design misses is time. We propose the use of discrete event system specification (DEVS) in the design and analysis of a software system, such as RCD. With DEVS, coupling between objects can be specified explicitly and an object behavior can be shown in time. We introduce DEVS, show the time-line analysis of Remote Consultation and Diagnosis session playback using DEVS, and then describe its implementation. PMID- 11020821 TI - A fundamental metric for continuity of care: modeling and performance evaluation. AB - The concept of continuity in medicine is fundamental and it refers to the delivery of medical care to a patient by a care provider in an uninterrupted and coordinated manner and in accordance with the medical care needs of the patient. Principal limitations of previous studies include the following. First, there is the absence of a deliberate effort to schedule a patient's successive visits with the same provider. Second, in most cases, the reported measures are derived from the data collected through questionnaires that patients and doctors are asked to complete from memory. This introduces an unreliability factor. Third, the reported continuity-related measures are restricted to the available patient sample and, as a result, they do not necessarily reflect a representative patient distribution by age in a typical community. Fourth, the relationship between the patient-doctor interaction interval and the continuity of medical care has never been explored systematically in the literature. This paper models continuity for a representative staff model health maintenance organization (HMO) clinic in suburban Arizona and simulates patient visits that are stochastically generated through utilizing representative numbers of patient visits and care providers. It focuses on individual-based continuity delivered by a primary care provider, i.e., a general practitioner. In the simulation, the visit patterns of patients, their ages, and the length of the interaction episodes are synthesized stochastically. The distributions reflect both the patient visit profile, by age, inherent in the 90,000+ patient electronic records collected at the CIGNA HMO clinic at Chandler, AZ, over a three-year period, and the population distribution inherent in the CIGNA records. A key characteristic of the model is that it aims at deliberately maximizing continuity by making a strong effort to schedule both a patient's regular visit and follow-ups with the primary care provider, subject to the provider's availability and schedule. This paper proposes a new definition of continuity, fundamental continuity of care index (FCCI), and argues that, fundamentally, a primary care provider's depth of understanding of the patient is directly proportional to the total length of interaction between the patient and the primary care provider. Utilizing the CIGNA summary data, this paper organizes patients into five age groups: 0-5 years, 5-15 years, 15-45 years, 45-60 years, and 60+ and synthesizes the corresponding arrival distributions. Performance results, obtained for a total of 55,056 patient visits over a three-year period indicate that, while 94-97% of the patient visits are with the primary care provider, patients spend 76-77% of the nominal total visit time with their primary care provider, leading to FCCI values ranging from 0.72 to 0.75. Performance results indicate that for different choices of the mean of the patient-doctor interaction duration, the average FCCI values, obtained following simulation, reveal an approximate bell-shaped graph. For values of the mean ranging from 5 to 10, 15, 20, 25, 35, 45, and 55 min, the average FCCI measure starts at 0.49, increases to a maximum of 0.88 corresponding to a mean of 25 min, and then decreases to 0.60. Thus, for a medical clinic, constrained by the number of care providers servicing a fixed number of patients, for a given population distribution by age, an optimum value for the average patient-doctor interaction duration may be obtained empirically that yields the maximal continuity measure. PMID- 11020822 TI - A communication server for telemedicine applications. AB - The telemedicine applications, in some cases, need multipoint-to-multipoint communication. To meet the requirement of telemedicine communication, the development of a medical communication server is proposed in this paper. To make a working, as well as cost-effective, communication platform for the telemedicine applications, a specially designed communication server model is proposed in this work. This server is able to provide an effective multipoint-to-multipoint communication service for any level applications in telemedicine. The implementation program of this server is developed in a Windows'95 environment by using a winsocket. The trial application testing in a telemonitoring system is also presented to demonstrate the feasibility of taking such a structure with the server. By using the architecture of such a communication server in telemedicine applications, the multipoint-to-multipoint communication is easily managed and the communication processes are simplified and well controlled by the server. PMID- 11020823 TI - Healthcare information technology: what is on the horizon? PMID- 11020824 TI - Telecommunications and the reform process in public health. PMID- 11020825 TI - Dynamic image data compression in spatial and temporal domains: theory and algorithm. AB - Advanced medical imaging requires storage of large quantities of digitized clinical data. These data must be stored in such a way that their retrieval does not impair the clinician's ability to make a diagnosis. In this paper, we propose the theory and algorithm for near (or diagnostically) lossless dynamic image data compression. Taking advantage of domain-specific knowledge related to medical imaging, the medical practice and the dynamic imaging modality, a compression ratio greater than 80:1 is achieved. The high compression ratios are achieved by the proposed compression algorithm through three stages: 1) addressing temporal redundancies in the data through application of image optimal sampling, 2) addressing spatial redundancies in the data through cluster analysis, and 3) efficient coding of image data using standard still-image compression techniques. To illustrate the practicality of the proposed compression algorithm, a simulated positron emission tomography (PET) study using the fluoro-deoxy-glucose (FDG) tracer is presented. Realistic dynamic image data are generated by "virtual scanning" of a simulated brain phantom as a real PET scanner. These data are processed using the conventional [8] and proposed algorithms as well as the techniques for storage and analysis. The resulting parametric images obtained from the conventional and proposed approaches are subsequently compared to evaluate the proposed compression algorithm. As a result of this study, storage space for dynamic image data is able to be reduced by more than 95%, without loss in diagnostic quality. Therefore, the proposed theory and algorithm are expected to be very useful in medical image database management and telecommunication. PMID- 11020826 TI - CT-based software for 3-D localization and reconstruction in stepping source brachytherapy. AB - This paper describes innovative software for catheter localization and three dimensional (3-D) reconstruction in stepping source brachytherapy applications. Patient information is a set of computed tomography (CT) slices scanned during the implantation of brachytherapy catheters. Catheter geometry and patient anatomy are exported for use with dose calculation software modules. The errors produced by the system are also encouragingly low. Time saving was achieved, in terms of other traditional reconstruction techniques. Various automated procedures, 3-D graphics and a user-friendly GUI, have contributed to providing a powerful, comprehensive software tool, directly useable in the clinical practice. PMID- 11020827 TI - A technique for extracting physiological parameters and the required input function simultaneously from PET image measurements: theory and simulation study. AB - Positron emission tomography (PET) is an important tool for enabling quantification of human brain function. However, quantitative studies using tracer kinetic modeling require the measurement of the tracer time-activity curve in plasma (PTAC) as the model input function. It is widely believed that the insertion of arterial lines and the subsequent collection and processing of the biomedical signal sampled from the arterial blood are not compatible with the practice of clinical PET, as it is invasive and exposes personnel to the risks associated with the handling of patient blood and radiation dose. Therefore, it is of interest to develop practical noninvasive measurement techniques for tracer kinetic modeling with PET. In this paper, a technique is proposed to extract the input function together with the physiological parameters from the brain dynamic images alone. The identifiability of this method is tested rigorously by using Monte Carlo simulation. The results show that the proposed method is able to quantify all the required parameters by using the information obtained from two or more regions of interest (ROI's) with very different dynamics in the PET dynamic images. There is no significant improvement in parameter estimation for the local cerebral metabolic rate of glucose (LCMRGlc) if the number of ROI's are more than three. The proposed method can provide very reliable estimation of LCMRGlc, which is our primary interest in this study. PMID- 11020828 TI - CAR special issue of the IEEE transactions on information technology in biomedicine. PMID- 11020829 TI - WebOnCOLL: medical collaboration in regional healthcare networks. AB - This paper presents WebOnCOLL, a web-based medical collaboration environment, which has been designed in the context of the regional healthcare network of Crete. WebOnCOLL employs the infrastructure of regional healthcare networks to provide integrated services for virtual workspaces, annotations, e-mail, and on line collaboration. Virtual workspaces support collaborative concepts like personal web pages, bulletin boards, discussion lists, shared workspaces, and medical case folders. Annotations provide a natural way for people to interact with multimedia content, while e-mail is one of the most popular forms of communication today. On-line collaboration satisfies the need for a more direct form of communication. PMID- 11020830 TI - Full-field direct digital telemammography: technical components, study protocols, and preliminary results. AB - The early detection of breast cancer increases the survival rate in women. Today, film-screen mammography is the most common and effective technique for the detection of breast cancer. However, the film-screen image recording system of current mammography has several technical limitations that can reduce the breast cancer diagnostic accuracy. A state-of-the-art technology, full-field direct digital mammography (FFDDM) has the potential to increase the sensitivity of clinical and screening examinations and emerge as a film-screen mammography replacement. It is believed that efficiently delivering the superior image quality of FFDDM to expert mammographers will significantly increase the breast cancer diagnostic accuracy. This concept motivates the studies of FFDDM telemammography. In this paper, we will report the FFDDM telemammography project that is being conducted in our facility. The fundamental technology of the FFDDM system and the characteristics of FFDDM images are described. Our research approach is through three protocols: telediagnosis, teleconsultation, and telemanagement. Each of these study protocols is defined. To conduct this FFDDM telemammography project, an asynchronous transfer-mode-network-based telemammography system is developed across two remote campuses in our facility. The architecture of this system is detailed. Unlike other medical imaging modalities, the matrix of an FFDDM image is huge. How to present the image information to mammographers via currently available display media is a challenging task. Our display approaches for the FFDDM images are described. Some preliminary study results from the current phase of this study are reported. PMID- 11020831 TI - An open medical imaging workstation architecture for platform-independent 3-D medical image processing and visualization. AB - A need for an entirely new medical workstation design was identified to increase the deployment of 3-D medical imaging and multimedia communication. Recent wide acceptance of the Word Wide Web (WWW) as a general communication service within the global network has shown how big the impact of standards and open systems can be. Information is shared among heterogeneous systems and diverse applications on various hardware platforms only by agreeing on a common format for information distribution. For medical image communications, the Digital Imaging and Communication in Medicine (DICOM) standard is possibly anticipating such a role. Logically, the next step is open software: platform-independent tools, which can as easily be transferred and used on multiple platforms. Application of the platform-independent programming language Java enables creation of plug-in tools, which can easily extend the basic system. Performance problems inherent to all interpreter systems can be circumvented by using a hybrid approach. Computationally intensive functions like image processing functions can be integrated into a natively implemented optimized image processing kernel. Plug-in tools implemented in Java can utilize the kernel functions via a Java-wrapper library. This approach is comparable to the implementation of computationally intensive operations in hardware. PMID- 11020832 TI - Voxel-based 2-D/3-D registration of fluoroscopy images and CT scans for image guided surgery. AB - Registration of intraoperative fluoroscopy images with preoperative three dimensional (3-D) CT images can be used for several purposes in image-guided surgery. On the one hand, it can be used to display the position of surgical instruments, which are being tracked by a localizer, in the preoperative CT scan. On the other hand, the registration result can be used to project preoperative planning information or important anatomical structures visible in the CT image onto the fluoroscopy image. For this registration task, a novel voxel-based method in combination with a new similarity measure (pattern intensity) has been developed. The basic concept of the method is explained at the example of two dimensional (2-D)/3-D registration of a vertebra in an X-ray fluoroscopy image with a 3-D CT image. The registration method is described, and the results for a spine phantom are presented and discussed. Registration has been carried out repeatedly with different starting estimates to study the capture range. Information about registration accuracy has been obtained by comparing the registration results with a highly accurate "ground-truth" registration, which has been derived from fiducial markers attached to the phantom prior to imaging. In addition, registration results for different vertebrae have been compared. The results show that the rotation parameters and the shifts parallel to the projection plane can accurately be determined from a single projection. Because of the projection geometry, the accuracy of the height above the projection plane is significantly lower. PMID- 11020833 TI - [Acute renal insufficiency due to vomiting or diarrhea: the Achilles heel of medical support for heart failure]. AB - Morbidity and mortality of heartfailure are decreasing because of improved medical treatment. The recompensation balance can however be very fragile, which is illustrated by three patients. A woman aged 73 and a man aged 62, both known with heart failure, became seriously ill after diarrhoea; examination revealed uraemia, hyperpotassaemia and raised digitalis levels. A man aged 72, also with heart failure and diarrhoea, developed sensory disorders in both feet. He, also, was found to suffer from renal insufficiency. All patients used an angiotensin converting enzyme inhibitor or an angiotensin-II-antagonist and an aldosterone receptor blocker as well. During the (innocent) intercurrent disease by which their intravascular volume diminished they developed a severe renal insufficiency which needed clinical, intensive treatment. After haemodialysis, all three recovered well. Renal function plays an essential role in the fragile treatment balance that exists in recompensated cardiac patients. The medication that is beneficial for the cardiac functioning may at the same time compromise the renal perfusion. Good monitoring and good instructions about what to do in case of intercurrent diseases are of vital importance in these patients. PMID- 11020834 TI - [Saint John's Wort as an antidepressant]. AB - Although in Germany St. John's wort (Hypericum perforatum) is the most widely prescribed antidepressant, in the Netherlands little is known about it. Nevertheless patients ask for it more often or take it as self-medication. There has been much research into the antidepressant efficacy of hypericum extracts, but the methodological quality has been moderate. In recent years studies with a much better design were published in European journals. Hypericum extract in a dosage of 900 mg/day is effective in mild to moderately severe depressed outpatients. Maintenance therapy has not been studied sufficiently yet. The side effect profile is on a placebo level. Recently interactions with other medications were described (decrease of plasma level/clinical efficacy of protease inhibitors, warfarin, theophyllin, cyclosporin, third generation oral contraconceptives, digoxin and phenprocoumon). A patient's request to be treated with hypericum extract can be considered, e.g. if the patient experiences troublesome side effects on other drugs. In case hypericum is the initial antidepressant and the effect falls short of the expectations, treatment with a synthetic standard antidepressant should be applied. PMID- 11020835 TI - [Roaming through methodology. XXIII. The need for randomization and blinding in therapeutic investigations]. AB - After the start of a treatment patients often get better. This may be due to the treatment or other circumstances. There may be errors in the execution of the study, the disease may disappear spontaneously, effective co-interventions may be present and possibly there are also placebo effects. Most of the time, only sufficiently large randomised, blinded research is useful to estimate the efficacy of drugs and of other treatments. The best comparison is often between the new treatment and the best available one, not the sham treatment. PMID- 11020836 TI - [Diagnostic image (3). (Lithopedion)]. AB - In a 62-year-old woman with aspecific symptoms in the lower abdomen a lithopaedion was diagnosed mimicking an ovarian tumour. PMID- 11020837 TI - [From gene to disease; mutations in interleukin-12-receptor-beta 1- and interferon-gamma-receptor-1 lead to nontuberculous mycobacterial infections and salmonellosis]. AB - In patients with severe and relapsing infections with non-tuberculous mycobacteria and Salmonella autosomal recessive as well as dominant-negative mutations are shown in genes for receptors of type-1 cytokines: the IL-12 receptor and the IFN-gamma receptor. In case of an impaired capacity to produce these cytokines or--given an abnormal receptor--to react to them, the cellular immune reaction does not run a normal course and the susceptibility to infection by intracellular pathogens is enhanced. PMID- 11020838 TI - [Incidence of herpes neonatorum in Netherlands]. AB - OBJECTIVE: Investigation of the incidence of neonatal herpes in the Netherlands between 1992 and 1998. DESIGN: Inventory questionnaire survey. METHODS: All virological laboratories in the Netherlands were sent a questionnaire on the number of culture proven cases of neonatal herpes recorded between 1992 and 1998 and on the type of herpes simplex virus (HSV-1 or HSV-2). The gynaecological and paediatric departments of all university hospitals and of half of the general hospitals were sent questionnaires as well. Gynaecologists were asked how often caesarean section was performed in order to prevent neonatal herpes and how frequently pregnant women were seen with genital herpes. Paediatricians were asked how often they observed neonatal herpes, the type of HSV and the possible transmission route. Based on these data the figures for the whole of the Netherlands were estimated. RESULTS: The incidence of neonatal herpes in the Netherlands in the period 1992 to 1998 was 2.4 per 100,000 neonates. HSV-1 was the cause of neonatal herpes in 73%, HSV-2 in 9%, and in 18% of the cases the type of infection was not recorded. The number of pregnant women with genital herpes had increased, but, in agreement with a consensus statement, the gynaecologists hardly performed caesarean sections any more to prevent neonatal herpes (2 per year). CONCLUSIONS: The incidence of neonatal herpes in the Netherlands had not increased. There was no predominant role of HSV type 2 causing neonatal herpes. PMID- 11020839 TI - [Food additives as a cause of medical symptoms: relationship shown between sulfites and asthma and anaphylaxis; results of a literature review]. AB - OBJECTIVE: To determine if a causal connection exists between food additives and various medical complaints. DESIGN: Literature study. METHOD: Medline over the period January 1966-January 1999 was searched for articles on the following substances not containing protein and lactose: monosodium glutamate (MSG), sulfites, azo-dyes (tartrazine, sunset yellow, azorubin, amarant, cochineal red), benzoates, sorbates, butylated hydroxyanisole/butylated hydroxytoluene (BHA/BHT), parabens, cinnamon and vanilla, in combination with key words regarding food and side effects. Of those studies purporting to demonstrate an effect, only double blind randomized placebo-controlled studies with oral challenge were assessed further, unless the complaint was anaphylaxis. Of studies not demonstrating an effect the design was assessed. RESULTS: Only for sulfites as causative agents of asthma and anaphylaxis, methodologically adequate studies demonstrating a causal connection could be found. For azo-dyes, benzoates, MSG, sorbates and BHA/BHT, no link with medical symptoms was demonstrable. For parabens, cinnamon and vanilla there were insufficient or inadequate data to justify a conclusion. PMID- 11020840 TI - [Neuroradiologic intervention in two patients with cerebral sinus thrombosis]. AB - Two patients with cerebral sinus thrombosis were successfully treated with neuroradiological intervention procedures, one with local thrombolysis and the other with mechanical thrombosuction using a hydrolyser catheter. The first patient, a 20-year-old woman, was treated with asparaginase for acute lymphatic leukaemia. She lapsed into coma with extensor posturing due to superior sagittal and right transverse sinus thrombosis. She recovered completely after local thrombolysis with 2,940,000 units urokinase, administered over a period of 40 hours. The second patient was a 29-year-old man who presented with clinical deterioration after seizures due to superior sagittal, left transverse and straight sinus thrombosis. A CT-scan demonstrated bilateral haemorrhagic cerebral infarctions. Since the risk of haemorrhage during thrombolysis with urokinase was considered to be high, mechanical thrombosuction with a hydrolyser catheter was performed. This procedure took only 4 hours. The patient recovered completely in two weeks. These cases add further evidence to the effectiveness of thrombolysis and thrombosuction in selected patients with severe cerebral sinus thrombosis. PMID- 11020841 TI - [Consensus for infliximab treatment of patients with Crohn's disease]. AB - Very recently infliximab--a tumour necrosis factor blocking antibody--was registered as an important novel immunomodulating therapy for active Crohn's disease in the Netherlands. Although this treatment can have a very significant and sometimes dramatic effect (on remission induction), its longterm effects are still uncertain and the high cost is a major drawback. An expert committee of specialists in inflammatory bowel disease (IBD) in the Netherlands therefore produced a consensus report on specific treatment indications for its use. It is stated that only immunosuppressive-resistant patients or patients with incompatibilities and allergies to prednisone, azathioprine and methotrexate should be candidates, a second indication being patients with serious enterocutaneous fistulae not reacting to a full course of conventional therapy. Its use is not indicated in patients with stenosing disease. It is also stated that the effect of longterm repeated therapy in active Crohn's disease still has to be established. Since budget limitations for medications are a major disadvantage, the formation of expert committees (with an expert in the treatment of complicated inflammatory bowel disease) in each hospital should be stimulated to limit the treatment to only cases with real immunosuppressive-resistant disease. PMID- 11020842 TI - [Clinical thinking and decision-making in practice. A patient with persistent fever]. PMID- 11020843 TI - [Meperidine (pethidine) outdated as analgesic in acute pancreatitis]. PMID- 11020844 TI - Totally implantable venous access device inserted during major abdominal operations for cancer. A new technique. AB - Totally implantable venous access devices are usually implanted by surgical cut down technique or by percutaneous approach, after a first surgical procedure. The authors describe a new surgical approach utilizing the right gonadal vein or the vena cava for totally implantable venous access device placement in patients submitted to major digestive surgical procedures for tumors. This new surgical approach allows one to avoid a second operative procedure. PMID- 11020845 TI - Pseudotumorous hyperplasia of the caudate lobe in the non-cirrhotic liver: MR and CT arterial portography appearance. AB - We describe the magnetic resonance image and computed tomographic arterial portography appearance of pseudotumorous hyperplasia in the caudate process of the caudate lobe in a non-cirrhotic patient. A prominent portal vein branch directly arising from the right main portal branch was seen in the center of the lesion on magnetic resonance imaging and computed tomographic arterial portography. We think that portal flow change of the caudate lobe by an anomalous portal branch may be correlated to pseudotumorous hyperplasia. PMID- 11020846 TI - The effect of microvesicular fatty change on liver regeneration after partial hepatectomy. AB - BACKGROUND/AIMS: Hepatic resection for primary and secondary liver tumors and transplantation of segments of liver into patients with severe liver disease has become a common practice. However, the information on the regenerative potential of liver with various underlying diseases, particularly, the common disorder fatty change, is sparse. METHODOLOGY: In the present study we have evaluated the regenerative potential of liver in mice lacking fatty acyl-CoA oxidase (AOX-/-) after partial hepatectomy. Liver regeneration was assessed by measuring mitotic activity and 5-bromo-2'-deoxyuridine (BrdU) labeling using immunohistochemistry. RESULTS: BrdU labeling index in AOX-/- mice was 99 +/- 32, 156 +/- 28, 54 +/- 13, and 120 +/- 33 per 1000 cells at 48, 60, 72 and 84 hours, respectively after partial hepatectomy. Comparable BrdU labeling index was observed in AOX+/+ mice. Similarly, mitotic index in both groups was comparable. CONCLUSIONS: These results suggest that DNA synthesis after partial hepatectomy in genetically altered mice with microvesicular fatty change is unaffected. PMID- 11020847 TI - A new case of congenital cyst of the pancreas. AB - A new case of congenital cyst of the pancreas is reported. A 34-year-old woman was admitted with a painless, large, epigastric mass. Ultrasonography revealed an anechogenic retrogastric tumor. Computed tomography scan described a liquid cyst of the pancreas which was 15 cm in diameter. During surgery, a well-delimited translucent cyst was found and no local malignancy was observed. Extensive distal pancreatic resection with preservation of the spleen was performed and a thin part of cephalic pancreas was preserved. The liquid of the cyst did not contain any mucus. Microscopic study of the cyst wall described normal cuboidal cells and congenital cyst of the pancreas was diagnosed. Several diagnoses including hydatid cyst, pseudocyst and cystic tumors of the pancreas are discussed. Before surgery, lack of acute pancreatitis in recent medical history rules out pseudocyst and hydatid cyst. During the operation, if cystadenocarcinoma is easily ruled out, macrocystic serous cystadenoma is more difficult to exclude. Only histological examination of the cystic wall confirms the difference between cystadenoma and congenital cyst which remains an exceptional entity. PMID- 11020848 TI - Association between common bile duct stones and treated hypothyroidism. AB - BACKGROUND/AIMS: The purpose of this study was to investigate the frequency of diagnosed hypothyroidism in patients with common bile duct stones. METHODOLOGY: The common bile duct stone group (Group I) consisted of all the patients who had verified gallstones in the common bile duct in endoscopic retrograde cholangiopancreatography during 1995. The control group (Group II) was matched for age, sex, and hospital admission. These patients did not have diagnosed gallbladder or common duct stones. In both groups there were 86 patients (56 women and 30 men). The median age in these groups was 73 (range: 22-92) years at the time of common bile duct stone diagnosis. Medical records of all patients were reviewed. RESULTS: In Group I the prevalence of previously diagnosed hypothyroidism was 7/86 (8%) compared with 1/86 (1%) in Group II (P = 0.01). Hypothyroidism was previously diagnosed only in the common bile duct stone patients (Group I) of over 60 years of age, where the prevalence was 7/66 (11%). In addition, we studied 36 consecutive gallbladder stone patients (Group III) of over 60 years, who had no evidence of common bile duct stones. Hypothyroidism had been diagnosed in them less frequently (2/36 = 6%) than in the age and hospital admission matched common bile duct stone patients (P = 0.01). Other diagnosed endocrine disorders did not differ between the study groups. CONCLUSIONS: There is a significant association between the common bile duct stones and previously diagnosed hypothyroidism. There stronger association between the common bile duct stones and hypothyroidism compared to gallbladder stones and hypothyroidism suggests a mechanism other than merely the cholesterol metabolism mediated mechanism. The 11% prevalence of previously diagnosed hypothyroidism in the common bile duct stone patients of over 60 years of age suggests all patients with common bile duct stones be screened for current thyroid dysfunction. PMID- 11020849 TI - Combined laparoendoscopic approach for biliary lithiasis treatment. AB - BACKGROUND/AIMS: The treatment of common bile duct stones diagnosed during videolaparoscopic cholecystectomy is still under debate. In cases of suspected common bile duct stones, a double approach with endoscopic retrograde cholangiopancreatography either prior to, or following videolaparoscopic cholecystectomy is the current routine in many centers. An intraoperative endoscopic retrograde cholangiopancreatography with endoscopic papillosphincterotomy and stone extraction has recently been proposed. METHODOLOGY: We compared the approaches for suspected common bile duct stones in 21 cases of combined intervention endoscopic retrograde cholangiopancreatography during videolaparoscopic cholecystectomy to 17 cases of sequential intervention (endoscopic retrograde cholangiopancreatography prior to videolaparoscopic cholecystectomy). Complications and postoperative monitoring are discussed and reported on the basis of hospital stay. RESULTS: Although the efficacy and the complications are similar, patients treated with the sequential approach stayed in the hospital longer because of the double monitoring period during both after endoscopic retrograde cholangiopancreatography and after videolaparoscopic cholecystectomy. CONCLUSIONS: A combined approach to suspected common bile duct stones during videolaparoscopic cholecystectomy could be an effective and a financially worthwhile treatment. PMID- 11020850 TI - Results of resection for hilar cholangiocarcinoma with analysis of prognostic factors. AB - BACKGROUND/AIMS: Resection for hilar cholangiocarcinoma remains a challenging procedure and recent published results continue to show that few patients are cured of their disease. The objective of this review was to determine the results of radical resection and to identify factors associated with long-term survival. METHODOLOGY: Retrospective review of resection for hilar cholangiocarcinoma in 29 consecutive patients with statistical analysis of prognostic factors, including p53 status. RESULTS: The mortality and morbidity rates were 6.9% and 34%, respectively. The overall 5-year survival was 20% with the median survival being 16 months. Univariate analysis identified the following factors associated with poor survival; involved lymph nodes, vascular invasion, advanced tumor stage, positive tumor margins, and p53 mutation. However, none of these factors was associated with poor survival by multivariate analyses. CONCLUSIONS: Radical resection for hilar cholangiocarcinoma can be performed with acceptable morbidity and mortality, but most patients succumb to their disease. p53 status may be a helpful adjunct to routine pathological staging. PMID- 11020851 TI - Palliative percutaneous transhepatic gallbladder drainage of gallbladder empyema before laparoscopic cholecystectomy. AB - BACKGROUND/AIMS: This study is conducted to evaluate the feasibility of percutaneous transhepatic gallbladder drainage prior to laparoscopic cholecystectomy for the treatment of gallbladder empyema. We also determine the sonographic findings, causative organism, clinical signs and symptoms, laboratory data, associated underlying medical disorders and the complications related to both cholecystostomy and laparoscopic cholecystectomy. METHODOLOGY: One hundred and forty-five cases of gallbladder empyema were included in this study which was composed of 80 males and 65 females, aged 22-94 years with a mean age of 71 years. All patients underwent percutaneous transhepatic gallbladder drainage under ultrasound and fluoroscopic guidance, and laparoscopic cholecystectomy was carried out thereafter. We analyzed the clinical presentations (signs, symptoms, laboratory and ultrasonographic findings, concomitant medical disorders), causative organisms and the complications related to percutaneous cholecystostomy and laparoscopic cholecystectomy. RESULTS: Percutaneous transhepatic gallbladder drainage was performed successfully in all patients within 48 hours after clinical diagnosis of acute cholecystitis. Complications related to percutaneous transhepatic gallbladder drainage were bile leakage after tract dilatation noted in 2 patients (1.4%), and 20 (14%) patients had pain at the puncture site which radiated to the right shoulder during the procedure, but resolved spontaneously within an hour later. On admission, 102 (70%) patients presented as right upper quadrant pain, 39 (27%) as epigastric pain, 90 (62%) as fever, 108 (74%) patients had leukocytosis, and 33 (22.7%) patients were septic. AST and ALT were elevated in 57% and 51% of patients, respectively. Alkaline phosphatase was elevated in 56% of patients, and 34% of those patients had combined common bile duct stones. Gallbladder stones were documented in 135 (93%) patients, while the remaining 10 (7%) cases were acalculous. Five (3.4%) patients had combined gallbladder adenocarcinoma, 7 (4.8%) had liver abscess, while 13 (9%) had biliary pancreatitis. The ultrasonographic findings included gallbladder distension (93%), wall thickening (90%), pericholecystic fluid accumulation (15%), intraluminal sludge or stone (93%) and intraluminal air (13.9%). Bile culture were positive in 83% of the cases and showed gram-negative bacteria in 75%, gram positive in 30%, anaerobes in 7%, while no growth in the remaining 17% of the cases. The common pathogens were Escherichia coli (57%), Enterococcus (27%), Klebsiella pneumonia (18%), Morganella morganii (7.6%), Pseudomonas aeruginosa (4.1%) and Salmonella (0.7%). The total postoperative complication rate was 17%, which included wound infection, bleeding, subhepatic abscess, cystic duct stump leak, common bile duct injury and pneumonia. Postoperative mortality was 2.6%. Conversion rate to open cholecystectomy was 27%. Clinical conditions improved within 48 hours after cholecystostomy in 93% of patients. Time interval between cholecystostomy and elective cholecystectomy was 2-21 days with a mean of 4 days. Total hospital stay was 5-38 days (mean: 11 days). CONCLUSIONS: Percutaneous transhepatic gallbladder drainage is a safe and effective procedure for the initial management of gallbladder empyema. We highly recommend this preoperative drainage procedure in patient with sepsis, and for those high-risk patients such as old age and with underlying medical illnesses. This procedure can stabilized the patient so that an appropriate therapeutic planning can be achieved. PMID- 11020852 TI - A simple novel method of biliary drainage: gallbladder serosal wrapping after open choledochotomy. AB - In this study, we developed a new, simple technique for biliary drainage after open choledochotomy of choledocholithiasis. After the absence of intraductal stones was established by operative cholangiography and cholangioscopy, preserved gallbladder serosal wrapping was performed by inclosing a polyethylene tube (C tube), which was inserted from the cystic duct to optimal portion of choledochus, within the gallbladder bed, with continuous suture of the preserved serosa of the gallbladder using 4-0 absorbable thread. This method was used in the cases of 8 patients. There was neither bile leakage nor residual bile duct stones. The C tube could be removed after 7 days following surgery. The average hospital stay was 12.3 +/- 6.6 days. We propose that this procedure would be very simple and useful, and it would significantly shorten hospital stays after open choledochotomy of choledocholithiasis. PMID- 11020853 TI - Gallbladder hematoma in a patient with hemophilia B, report of a case. AB - The authors report the case of a 49-year-old man, hematoma of the gallbladder associated with hemophilia B. The patient was diagnosed incidentally by abdominal computed tomography with a gallbladder tumor. Endoscopic retrograde cholangiopancreatography revealed negative gallbladder and angiography showed no obvious abnormality in the cystic artery. Magnetic resonance imaging showed a mass of mixed intensity on T2-weighted image and a mass of mixed intensity on the lining of the gallbladder wall. Hemorrhage in the gallbladder was thought to be most likely, however, the gallbladder tumor could not to be neglected. Cholecystectomy was performed on the patient and the pathological diagnosis was of a hemorrhage in the gallbladder. Hematoma of the gallbladder is a rare complication in patients with hemophilia B, however, it leads sometimes to a fatal course and magnetic resonance imaging is very useful to diagnose hemorrhage in the gallbladder. PMID- 11020854 TI - Composite neuroendocrine and adenocarcinoma of the common bile duct associated with Clonorchis sinensis: a case report. AB - The biliary tract has neuroendocrine cells of endoderm origin similar to the gastrointestinal tract, however neuroendocrine tumors of the biliary tract are rare. We report a composite glandular-endocrine cell carcinoma of the common bile duct in a 64-year-old Korean man which was associated with Clonorchis sinensis. The patient complained of right upper quadrant abdominal pain. Several parasites of Clonorchis sinensis were removed during the percutaneous transbiliary drainage. Endoscopic retrograde cholangiopancreatography revealed a polypoid mass (3 x 3 cm) with central ulceration in the common bile duct. Pancreaticoduodenectomy was performed. Microscopic examination of the tumor revealed a composite small cell neuroendocrine carcinoma and adenocarcinoma. The small cell carcinoma component showed positive reaction to chromogranin A and neuron-specific enolase and it was located mainly in the deeper portion of the mass. The well-differentiated adenocarcinoma component showed a positive reaction to carcinoembryonic antigen and it was situated in the superficial portion of the mass. Exclusively, the small cell component metastasized to the lymph node. It is suggested that this tumor could arise from a multipotential stem cell and showed neuroendocrine and glandular differentiation and that Clonorchis sinensis could be a predisposing factor, as in cholangiocarcinoma. PMID- 11020855 TI - Hepatopancreatoduodenectomy for squamous cell carcinoma of the gallbladder. AB - Herein we present a case of a 57-year-old woman with squamous cell carcinoma of the gallbladder, with an advanced, large tumor infiltrating the liver and duodenum, but curatively resected by hepatopancreatoduodenectomy. She has been doing well without any sign of recurrence more than 5 years after operating. Hepatopancreatoduodenectomy is an adequate procedure for en bloc resection of advanced squamous cell carcinoma of the gallbladder because it often grows expansively. No residual tumor cells owing to hepatopancreatoduodenectomy shall fortunately result in a good prognosis. PMID- 11020856 TI - Chemotherapy for gallbladder carcinoma--a surgeon's perspective. AB - Surgery is the only curative treatment for gallbladder cancer. However, because of frequent local-regional and distant recurrences, radical surgery during advanced stages of the disease is often unsuccessful. Despite potential improvements in patient survival and quality of life, alternatives such as chemotherapeutic regimens have not been fully developed for treatment of gallbladder cancer. The present paper reviews current publications that deal exclusively with the use of chemotherapy for treatment of gallbladder cancer, with particular attention to adjuvant chemotherapy including postoperative and preoperative systemic or regional hepatic arterial infusion chemotherapy. Treatment outcomes, including response rate, adverse effects, and survival are summarized. PMID- 11020857 TI - Characteristics of sarcomatous cholangiocarcinoma of the liver. AB - BACKGROUND/AIMS: The aim of this study was to clarify the clinicopathological characteristics of intrahepatic cholangiocarcinoma with sarcomatous changes. METHODOLOGY: Four cases of cholangiocarcinoma with sarcomatous change were identified and investigated. The clinicopathological findings, including the results of immunohistochemical staining, were investigated in comparison with those of ordinary cholangiocarcinoma. RESULTS: Two of them exhibited pyrexia as the initial symptom. The serum alkaline phosphatase level in sarcomatous cholangiocarcinoma was significantly lower than that in ordinary cholangiocarcinoma. Both the serum carcinoembryonic antigen and carbohydrate 19-9 level in sarcomatous cholangiocarcinoma also tended to be lower than those in ordinary cholangiocarcinoma. The carcinomatous component of all tumors was mostly poorly differentiated adenocarcinoma. The associated microscopic findings were as follows: lymphocyte infiltration in the tumor, accompanied by both necrosis and extensive lymph node metastases. Three of them predominantly exhibited spindle shaped sarcomatous changes, whereas the other case predominantly demonstrated pleomorphic-type sarcomatous changes. The sarcomatous area was positive for both vimentin, a mesenchymal marker, and for epithelial markers, furthermore, in 3 of 4 cases, the carcinoma portions were also positive for vimentin. The overall survival curves were not significantly different between the 2 groups, however, no long-term survivor was found in sarcomatous cholangiocarcinoma. CONCLUSIONS: The main characteristics of cholangiocarcinoma with sarcomatous changes are considered to be as follows: 1) often demonstrating pyrexia as a symptom; 2) not always demonstrating remarkable abnormal findings in the laboratory data including tumor markers; 3) histologically showing poorly differentiated adenocarcinoma; and 4) showing a very poor prognosis (especially, in a pleomorphic-type). PMID- 11020859 TI - Imaging-based dynamic liver testing--a computerized simulation. AB - BACKGROUND/AIMS: The mechanisms for maintaining liver function are complex and currently used liver function tests give limited and often nonspecific and insensitive results. The aim of the study was to demonstrate a dynamic liver test with a tomographic imaging modality as sampling method by means of a computer simulation. METHODOLOGY: A virtual three-dimensional liver model, consisting of parenchyma and bile ducts, was created using computer aided design/computer aided manufacturing software. An intravenously administered, bile excreted test substance was simulated in the model by awarding changing densities to the parenchyma and bile ducts in 12 identical sets of the model. RESULTS: Imaging based density measurements enabled the creation of time-density curves reflecting the transport of the simulated test substance in the parenchymal mass and bile ducts. By means of quantitative analysis of the curves, parenchymal uptake and excretion and intrahepatic bile flow could be assessed independently. CONCLUSIONS: The method enables comparison of function in different liver segments and may have particular value in investigating diseases that affect the liver in a non-homogeneous fashion, for example primary sclerosing cholangitis. Imaging sampling can theoretically be applied with any tomographic imaging technique for which a suitable test substrate exists, including computed tomography, magnetic resonance imaging or single photon emission tomography. PMID- 11020858 TI - Apoptosis and cell proliferation in the metaplasia-dysplasia-carcinoma-sequence of Barrett's esophagus. AB - BACKGROUND/AIMS: The regulation of apoptosis as a distinctive form of cell death and proliferation in the process of carcinogenesis in Barrett's esophagus is poorly understood. To investigate regulation of apoptosis, proliferation and the participation of the tumor suppressor gene p53, we examined these parameters in Barrett's metaplasia, dysplasia, and adenocarcinoma. METHODOLOGY: Apoptotic cells were identified and quantified in tissue specimens of 45 patients with different stages of Barrett's esophagus and normal fundus epithelium, respectively, using the in situ end-labeling and electron microscopy method in combination with morphological criteria. The tumor suppressor gene p53 was examined by direct sequencing of exon 4-8 as well as immunohistochemically. The proliferative activity was assessed by Ki67 immunostaining. RESULTS: Apoptotic cell death, identified by the in situ end-labeling and ultrastructural technique was significantly increased in Barrett's epithelium with intestinal metaplasia than in specimens with normal fundic epithelium and Barrett's carcinomas (P < 0.01). The proliferative activity, defined as Ki67 labeling index, was highest in adenocarcinomas (P < 0.01). P53 mutations were found in 8/9 adenocarcinomas and 2/5 specimens with dysplasia. Apoptosis was lower in p53 positive specimens of the metaplasia-dysplasia-carcinoma-sequence than in p53 negative specimens of Barrett's esophagus (P < 0.05). CONCLUSIONS: The higher levels of both apoptosis and proliferation indicate an increased cell turnover in Barrett's epithelium. Apoptosis seems to maintain tissue homeostasis, which is regulated by p53, and gradually lost in the metaplasia-dysplasia-carcinoma-sequence of Barrett's esophagus. PMID- 11020860 TI - p53 protein expression in intraductal papillary mucinous tumors (IPMT) of the pancreas as an indicator of tumor malignancy. AB - BACKGROUND/AIMS: Intraductal papillary mucinous tumors, as a cystic disease in the pancreas, clinically has a more indolent and favorable course than invasive ductal pancreas carcinoma. However, some cases of intraductal papillary mucinous tumors show invasive and rapid progression like ductal pancreas carcinoma and the prognosis of such patients is sometimes poor. In the current study, we carried out immunohistochemical staining of intraductal papillary mucinous tumor tissues for p53 and investigated whether positive staining indicates tumor malignancies and has a prognostic value for intraductal papillary mucinous tumors. METHODOLOGY: Nineteen (19) patients who underwent pancreatic resection under the diagnosis of intraductal papillary mucinous tumors at the Chiba University Hospital between April 1992 and December 1996 were studied. We performed immunohistochemical staining of p53 as well as of PCNA, Ki-67 and Bcl-2 using their respective antibodies. Pathological findings revealed that 9 cases were intraductal papillary adenoma, 9 were intraductal papillary adenocarcinoma, and one was invasive ductal papillary adenocarcinoma. RESULTS: p53 expression could only be detected in the 1 case with invasive ductal papillary adenocarcinoma. Significant association could not be found between histological features and immunohistochemical staining of PCNA, Ki-67 and Bcl-2. CONCLUSIONS: p53 protein expression could be detected after progression to invasive type of intraductal papillary mucinous tumors. The present results demonstrate that p53 expression might be an indicator of invasive progression in intraductal papillary mucinous tumors, and might represent a surgical indicator of intraductal papillary mucinous tumors. PMID- 11020862 TI - Gaucher's disease: a review for the internist and hepatologist. AB - Gaucher's disease is the autosomally recessively inherited deficiency of the lysosomal enzyme glucocerebrosidase. Increasing storage of glucocerebrosides leads to a multi-system disease which prevalence ranges between 1:30,000 and 1:50,000 in most countries. Thus only a minority of physicians are aware of this diagnosis, of the symptoms that should lead to its consideration, and of the availability of specific tests that confirm it. Because Gaucher's disease often affects the liver, hepatologists may care for Gaucher patients. This review provides the internist and hepatologist with practical information about recent advances in the management of the non-neuronopathic type I of Gaucher's disease. Gaucher's disease, type 1 should be considered when unexplained spleno- and hepatomegaly, anemia, thrombocytopenia, or skeletal disease are present, particularly in combination. The diagnosis is established by an assay for glucocerebrosidase activity in peripheral leukocytes. Lack of awareness and of widespread availability of the enzyme assay has as yet limited its application in clinical practice, and led to many cases of Gaucher's disease being diagnosed by bone marrow and liver biopsy. Alglucerase, placental enzyme preparation of glucocerebrosidase, has proven effective in more than 1,000 patients worldwide. Recently, alglucerase has been exchanged by the recombinant enzyme preparation imiglucerase, which is equally effective and safe. Enzyme replacement improves hematological abnormalities, hepato-splenomegaly, and quality of life in a matter of a few months. Regression of skeletal complications is usually seen only after 3-4 years. Recently gene therapy trials, which center on autotransfusion of retrovirally transduced stem cells, have successfully been started. PMID- 11020861 TI - Production and characterization of immortalized rat hepatocytes secreting hepatocyte growth factor/scatter factor. AB - BACKGROUND/AIMS: Hepatocyte transplantation is a recently attractive field in the treatment of liver failure and enzyme-deficient diseases. However, procurement of sufficient quantities of hepatocytes is almost impossible. We attempted to create a hepatocyte cell line that could be used for hepatocyte transplantation. METHODOLOGY: L2A2 is a conditionally immortalized rat hepatocyte cell line produced by transfection of temperature-sensitive simian virus T antigen to the hepatocytes in the Lewis rat. Hepatocyte Growth Factor/Scatter Factor (HGF/SF) secreting L2A2 cells, designated as SF-21, was produced by transfecting human HGF/SF cDNA into L2A2 cells. RESULTS: This cell line was able to produce HGF/SF at the rate of 5-10 ng/10(6) cells/24 hrs, and the recombinant HGF/SF was of the expected size and was functionally active in that it could scatter Madin-Darby canine kidney cells. The SF-21 cells grew faster than its parental cell clone, and survived and proliferated at 37 degrees C in vitro. Also, the SF-21 cells were able to survive and proliferate when transplanted into the spleen of syngeneic rat, and expressed glucose-6-phosphatase. CONCLUSIONS: These HGF/SF secreting hepatocytes can be used as a model system to test a feasibility of using genetically engineered hepatocyte cell line for hepatocyte transplantation in the rat. PMID- 11020863 TI - Expression of carbonic anhydrase I or II and correlation to clinical aspects of colorectal cancer. AB - BACKGROUND/AIMS: This study was undertaken to elucidate the correlation between the expression of carbonic anhydrase I or II and the characteristic features of colorectal cancer. METHODOLOGY: The carbonic anhydrase I or II expressions of 74 colorectal cancer patients were analyzed by Western blotting. The relative intensity of cancer to the paired normal mucosa was calculated, and then compared with the clinicopathological parameters. Furthermore, a multivariate analysis for synchronous distant metastasis was undertaken. RESULTS: The expression of carbonic anhydrase I in colon cancer or carbonic anhydrase II in rectal cancer with Duke's D was found to be significantly lower than that with Duke's B or C, respectively. Similarly, carbonic anhydrase I in colon cancer or carbonic anhydrase II in rectal cancer with moderate-severe budding was found to be significantly lower than that with none-mild budding, respectively. Based on the findings of a logistic regression analysis, carbonic anhydrase I was adopted for colon cancer (P = 0.057) and carbonic anhydrase II for rectal cancer (P = 0.008) regarding synchronous distant metastasis. CONCLUSIONS: The expressions of carbonic anhydrase I and II correlated with biological aggressiveness of colorectal cancer and synchronous distant metastasis, especially carbonic anhydrase I for colon cancer and carbonic anhydrase II for rectal cancer. PMID- 11020864 TI - Effect of blood transfusions on disease-free interval after rectal cancer surgery. AB - BACKGROUND/AIMS: To assess retrospectively the impact of perioperative blood transfusions on disease-free interval after curative surgery of rectal cancer. METHODOLOGY: One hundred and seven (107) subjects (65 men and 42 women, median age: 67) underwent anterior resection of the rectum for rectal cancer between 1990 and 1996. Thirty-six (37%) were transfused perioperatively. Outcome variables measured were: age, sex, distance of the tumor from the anal verge, type of stapled anastomosis, protective colostomy, cancer-free margin in the specimen, grading and staging, leak of the suture line, transfusion, number of blood units given, and adjuvant therapy. RESULTS: Transfusion, nodes involvement and adjuvant therapy had a significant impact on disease-free interval at the univariate analysis. The estimated probability of cancer-free survival at 5 years was 81% for the not transfused, and 63% for the transfused patients (P < 0.05). At multivariate analysis, nodes involvement resulted significantly and independently related to time to recurrence (P < 0.05) but also perioperative blood transfusion showed an important role as independent risk factor for rectal cancer relapse (P = 0.05). CONCLUSIONS: This study suggests that blood transfusion is likely to have a detrimental effect on disease-free interval after resective surgery of rectal cancer. PMID- 11020865 TI - Correlation between tissue levels of carboxy-terminal propeptide of type I procollagen and clinicopathological characteristics in human colorectal cancer. AB - BACKGROUND/AIMS: Type 1 collagen is the most abundant protein in the human body and a major constituent of the interstitial connective tissue. However, little is known about carboxy-terminal propeptide of type I procollagen (PICP) expression in human colorectal tumor tissues. We therefore evaluated the concentrations of PICP in colorectal tumor tissue as well as surrounding normal tissues and examined the relationship between its level and clinicopathological variables. METHODOLOGY: Tumor and normal tissues from 40 patients with colorectal carcinoma who had been operated on were stored at -80 degrees C until assays. PICP was assayed by sandwich immunoassay. RESULTS: It was found that the PICP level was significantly higher in the tumor extracts than in the normal tissue extracts (P < 0.0001). The ANOVA test showed that the level of PICP in tumor tissue was higher in the patients with advanced colorectal tumor than those with early stage disease (P < 0.0001). There were statistically significant differences with regard to the depth of tumor invasion, presence of lymph node metastasis, and hepatic metastasis (P < 0.05). There were also quantitative differences with respect to the PICP levels between obstructing tumors and non-obstructing carcinoma (P < 0.05), but the elevated PICP levels in tumor tissues were not associated with the type of histologic differentiation (P > 0.05). CONCLUSIONS: Although these findings suggest that PICP value in tumor tissue is most likely related to the some histomorphological architecture of the tumor, the authors believe that the prognostic significance of PICP expression in primary colorectal tumor and normal tissues requires further evaluation. PMID- 11020866 TI - The detection of colorectal cancer at an asymptomatic stage by screening is useful. AB - BACKGROUND/AIMS: The validity of mass screening using fecal occult blood testing remains controversial. In addition, no controlled clinical study has yet been performed to show the usefulness of sigmoidoscopy. The purpose of the present study was to compare the surgical results achieved in asymptomatic patients with colorectal cancer detected by screening with those in symptomatic individuals. METHODOLOGY: A total of 285 patients underwent a surgical resection of colorectal cancer between 1991 and 1997 at our institution. Among them, 233 patients had complaints related to cancer at the time of diagnosis. In contrast, 52 were asymptomatic. In those 52 patients, colorectal cancer had been suspected based on routine screening including fecal occult blood testing, colonoscopy and/or elevated serum levels of carcinoembryonic antigen. RESULTS: Early stage of colorectal cancer was more frequently seen in asymptomatic patients than in symptomatic patients P < 0.01. The survival rates for asymptomatic patients was also superior to those of symptomatic patients P < 0.05. CONCLUSIONS: Screening using fecal occult blood testing, colonoscopy and tumor markers is thus considered to be beneficial for the early detection of colorectal carcinoma, which also tends to demonstrate good surgical results. PMID- 11020867 TI - Mucosal antioxidant defense is not impaired in ulcerative colitis. AB - BACKGROUND/AIMS: In this study, mucosal antioxidant defense was investigated in the biopsy samples from 12 patients with active ulcerative colitis and from 13 patients under remission. METHODOLOGY: Biopsy samples obtained from healthy colon parts of the same subjects were used as control. RESULTS: No changes were observed between superoxide dismutase, glutathione peroxidase and catalase enzyme activities of control or inflamed biopsy samples. However, antioxidant potential values were found to be higher and malondialdehyde levels lower in inflamed samples compared with controls. CONCLUSIONS: Our results show that in contrast to previous suggestions, mucosal antioxidant defense is not impaired in ulcerative colitis. PMID- 11020868 TI - Rectal carcinoma with stercoral ulcer perforation. AB - We report a case of ruptured stercoral ulceration due to chronic constipation which is caused by rectal carcinoma. This case suffered from difficulty of stool passage for 5 months. Periumbilical pain and current-jelly stool were experienced before his admission. Physical examination revealed diffuse abdominal rebounding pain and laboratory data showed leukocytosis. Computed tomography demonstrated marked dilatation of the sigmoid colon with stool impaction due to neoplastic growth in the rectosigmoid junction. Thickening and edematous change of the colonic wall were noted. There was amorphous material with gas in the mesocolon, which indicated fecal peritonitis. Emergent operation with Hartman's procedure and left colostomy was performed. Diffuse pressure gangrene of the sigmoid colon wall with a perforating hole was identified. Pathologically, the resected colon specimen showed non-specific-acute and chronic inflammatory change. The perforating hole was surrounded by a necrotic border of ulcerative mucosa. After the operation, pelvic drainage was undertaken for 1 month and then the patient was discharged uneventfully. PMID- 11020869 TI - Chemotherapy-induced apoptosis of lymphocytes in esophageal cancer worsens outcome. AB - BACKGROUND/AIMS: Chemotherapy has been shown to induce apoptosis in esophageal cancer. However, no windows of opportunity exist to selectively kill tumor cells without killing host cells. Due to the concern that tumor-infiltrating lymphocytes may be killed by chemotherapy, we examined the significance of the effect of treatment on the density of tumor-infiltrating lymphocytes and apoptosis in the tumor itself and in the tumor-infiltrating lymphocytes. METHODOLOGY: In 93 patients with esophageal cancer including 50 with neoadjuvant chemotherapy, esophagectomy specimens were examined for density of tumor infiltrating lymphocytes and for apoptosis in both tumor cells and tumor infiltrating lymphocytes. RESULTS: Apoptotic index was increased by neoadjuvant chemotherapy only in tumor-infiltrating lymphocytes, apoptotic index was > or = 4 only in chemotherapy patients. The density of tumor infiltrating lymphocytes was a significant positive prognostic factor in chemotherapy and non-chemotherapy groups, and the high apoptotic index in tumor-infiltrating lymphocytes was an independent negative prognostic factor in the chemotherapy group. CONCLUSIONS: Apoptosis in tumor-infiltrating lymphocytes was induced by chemotherapy in some patients in association with a poor prognosis. Unexpectedly, chemotherapy did not increase apoptosis in tumor cells. Both findings suggest a need for improved regimes and individualized treatment. PMID- 11020870 TI - Esophageal anastomosis following gastrectomy for gastric cancer: comparison of hand-sewn and stapling technique. AB - BACKGROUND/AIMS: During the past 4 years, we have experienced a marked reduction in the incidence of esophageal anastomotic leakage and/or stricture coinciding with the use of a mechanical circular stapler for gastric cancer patients. METHODOLOGY: We reviewed medical records of gastric cancer patients who underwent a total or proximal gastrectomy, and compared the leakage or stricture of stapled anastomosis with the hand-sewn anastomosis technique. A total of 390 esophageal anastomosis were performed between January 1978 and August 1997. Two types of mechanical circular staplers were used (EEA and CDH). RESULTS: Anastomotic leakage occurred in 13 (3.3%) of 390 cases; three (4.5%) of 66 cases with hand sewn anastomosis, and 10 (3.1%) of 324 cases with stapled anastomosis (EEA: 4.5%, CDH: 0%). The anastomotic leakage rate was significantly lower in the CDH stapler group than in the EEA stapler group. Anastomotic stricture occurred in one (1.5%) of 66 cases of hand-sewn anastomosis, and 16 (4.9%) of 324 cases of stapled anastomosis (EEA: 5.9%, CDH: 2.9%). There were no significant differences in the stricture rate between the hand-sewn group and the stapler group. CONCLUSIONS: Stapling anastomosis using a CDH stapler led to a reduction in the incidence of anastomotic leakage. Surgeons must, however, continue to be aware of anastomotic stricture. PMID- 11020871 TI - Effect of rectal distension on the small intestine with evidence of a recto enteric reflex. AB - BACKGROUND/AIMS: To study the effect of rectal distension on jejunal and ileal motility aiming at the assessment of the possible role of rectal distension induced by constipation on the transport of the material in the gut. METHODOLOGY: The rectum of 16 healthy volunteers (mean age: 38.6 +/- 11.7 years, 10 men, and 6 women) was distended by a balloon filled with water in increments of 50 mL up to 200 mL and the response of the jejunal and ileal pressures was recorded. The test was repeated distending the anesthetized rectum 20 min and 3 hours after anesthetization. RESULTS: Rectal distension with 50 mL of water effected no jejunal or ileal pressure changes (P > 0.05). One hundred-mililitre (100-mL) rectal distension produced decrease of jejunal and ileal pressures (P < 0.05) which lasted as long as distension was maintained. Rectal distension with 150 and 200 mL caused jejunal and ileal pressure response similar to that of the 100 mL distension (P > 0.05). Distension of the anesthetized rectum effected no significant jejunal or ileal pressure changes. CONCLUSIONS: The results were reproducible in the individual subject. The decline of the intestinal pressure upon rectal distension postulates a reflex relationship between the 2 conditions. This reflex nature is evidenced by reproducibility and by its absence on distension of the anesthetized rectum. We termed this reflex relation: "recto enteric reflex". It is suggested that under normal physiologic conditions the reflex inhibits the intestinal transit, thus giving the rectum time to evacuate itself. Continuous rectal distension, as occurs in inertia constipation, appears to effect enteric hypotonia, a hypothesis which requires further studies. PMID- 11020872 TI - Octreotide prevents postoperative adhesion formation by suppressing peritoneal myeloperoxidase activity. AB - BACKGROUND/AIMS: Despite a great deal of effort to prevent the adhesion formation, an ideal therapy or drug remains to be identified. The aim of this study was to investigate the effect of octreotide on adhesion prevention and neutrophil infiltration. METHODOLOGY: Fourteen female Sprague-Dawley rats were subjected to a standard intraabdominal adhesion model operation. Octreotide group (n = 7) rats were administered 20 micrograms/kg octreotide intramuscular while control group (n = 7) rats were given no treatment. On the 10th day the rats were decapitated and the degrees of adhesions were recorded and the plasma, peritoneal fluid, peritoneal tissue myeloperoxidase levels were determined. RESULTS: Intraabdominal adhesions and peritoneal tissue myeloperoxidase levels were significantly reduced in the octreotide group as compared to saline treated adhesion group. CONCLUSIONS: We concluded that octreotide reduces the intraabdominal adhesions. This adhesion-preventive effect may be due to the inhibition of neutrophil infiltration. PMID- 11020873 TI - Intestinal permeability increases with the severity of abdominal trauma: a comparison between gas liquid chromatographic and enzymatic method. AB - BACKGROUND/AIMS: Increased intestinal permeability was found in humans after multiple trauma, burn injury and major vascular surgery. However, no data are reported regarding possible correlation between trauma and intestinal permeability degree. This study was undertaken to compare gas-liquid chromatographic and enzymatic method for the evaluation of intestinal permeability impairment in patients after severe abdominal trauma. METHODOLOGY: Five traumatized patients with an injury severity score of more than 24 and 5 cross-matched healthy volunteers were studied. The intestinal permeability was performed using a test solution, containing 10 g lactulose and 5 g mannitol. Gas liquid chromatographic method was applied to measure sugar standards and 5-hour urine samples and the results were compared with those obtained employing a specific enzymatic method. RESULTS: Linearity of myoinositol, lactulose and mannitol measured by gas-liquid chromatographic method was from 0.2-1 microgram injected. Using the enzymatic method, the response was linear between mannitol concentrations of 0.34 and 5.49 mM. Linearity of lactulose standard was from 0.18 2.92 mM. The gas-liquid chromatographic and enzymatic methods showed a good agreement using the Bland-Altman procedure. The mean lactulose/mannitol ratio was 0.085 +/- 0.025 in patients and 0.009 +/- 0.001 in controls (P < 0.001). The higher the injury severity score (30.8 +/- 5) the larger the ratio of lactulose to mannitol (R2 = 0.74). CONCLUSIONS: The enzymatic method--inexpensive, easy-to perform and timesaving--is suitable for intestinal permeability studies. An abdominal trauma, without injury requiring surgical operation, modifies the intestinal mucosa permeability possibly favoring passage of bacteria and subsequent sepsis. PMID- 11020874 TI - Endoscopic hemostasis for bleeding peptic ulcer using a hemostatic clip or pure ethanol injection. AB - BACKGROUND/AIMS: Endoscopic hemostasis is frequently chosen as the treatment of first choice for bleeding peptic ulcers. We retrospectively compared the hemostatic effects of hemoclips and endoscopic ethanol injection in patients with bleeding gastric and duodenal ulcers. METHODOLOGY: The subjects were 149 patients with 156 bleeding peptic ulcer lesions who were treated by endoscopic hemostasis, comprising 127 gastric ulcers and 29 duodenal ulcers. Hemoclips were used for 68 lesions (hemoclip group) and ethanol injection was done for 88 lesions (ethanol group). RESULTS: The hemostasis rates were 98.5% for the hemoclip group and 92.0% for the ethanol group. There was no significant difference in hemostatic effect between these two methods. All patients with unsuccessful hemostasis had a visible vessel larger than 2.0 mm in diameter and/or concomitant disease. CONCLUSIONS: The results suggest that these two endoscopic hemostatic methods are both highly effective. The presence of a large visible vessel may be predictive of unsuccessful hemostasis. PMID- 11020875 TI - Tauroursodeoxycholic acid administration as adjuvant therapy in cirrhotic patients on transplantation waiting lists. AB - BACKGROUND/AIMS: The lack of organ availability and an increased number of end stage cirrhotic patients has led to the lengthening of liver transplantation waiting lists. The progressive worsening of clinical and functional performance in patients awaiting the graft is one of the factors implicated in the increased mortality during the wait and in poor transplantation outcome. In this work our aim was to evaluate the effects of tauroursodeoxycholic acid administration on biochemical, clinical and functional parameters in a group of cirrhotic patients consecutively placed onto our liver transplantation waiting list. METHODOLOGY: Ten cirrhotic patients underwent biochemical, clinical and functional evaluation at the time of entering on our liver transplantation waiting list, then tauroursodeoxycholic acid was administered until liver transplantation. Complete evaluation was repeated every 2 months. The results were compared to those of a comparable historical control group that had undergone liver transplantation the year before the study. RESULTS: All patients were transplanted within 6 months from insertion on the waiting list. Longitudinal analysis of the treated group showed that cholestasis and cytolisis parameters constantly decreased and that gamma-glutamyl transpeptidase was significantly lower compared to baseline values at the 4th month of therapy. Clinical and functional parameters remained stable during follow-up. Comparison with the control group showed that gamma-glutamyl transpeptidase, alkaline phosphatase and both aminotransferases were reduced at the 4th month of therapy. Fewer days of hospital stay and less intensive care were required in the treated group. CONCLUSIONS: Treatment of end-stage cirrhotic patients awaiting liver transplantation with tauroursodeoxycholic acid improves biochemical parameters of cytolisis and cholestasis, and furthermore helps to maintain clinical and functional stability during the wait. Improved biochemical conditions and steady clinical-functional performance may promote better short term transplant outcome. PMID- 11020876 TI - Levels of interleukin (IL)-6, IL-8, and IL-1 receptor antagonist in the hepatic vein following liver surgery. AB - BACKGROUND/AIMS: The liver produces various cytokines, but local changes in the concentrations of these reaction products after liver surgery are unknown. We investigated the local changes of interleukin-6, interleukin-8 and interleukin-1 receptor antagonist after liver surgery. METHODOLOGY: We determined levels of interleukin-6, interleukin-8, and interleukin-1 receptor antagonist in the hepatic vein and radial artery after liver resection in 13 patients. These cytokine levels in the portal vein were also measured in 6 patients. RESULTS: Interleukin-6, interleukin-8, and interleukin-1 receptor antagonist levels were significantly increased during liver surgery (P < 0.05). The level of interleukin 6 was significantly lower in the hepatic vein than in the radial artery as well as in the portal vein at the end of the operation (P < 0.05, < 0.03). The level of interleukin-8 and interleukin-1 receptor antagonist was significantly higher in the hepatic vein than in the artery (P < 0.05). CONCLUSIONS: Interleukin-6 may be taken up by the liver after liver surgery, and the difference between hepatic venous and peripheral arterial interleukin-6 levels may be an indicator of liver regeneration after liver resection. Interleukin-8 and interleukin-1 receptor antagonist appear to be produced in the remaining liver. PMID- 11020877 TI - Intracystic infection of liver hydatidosis. AB - BACKGROUNDS/AIMS: The aim of this study was to review the clinical presentation and management of patients with intracystic bacterial infection in liver hydatidosis. METHODOLOGY: The records of 480 patients suffering from liver hydatid disease treated at our institution were reviewed. Only 42 patients fulfilled intracystic bacterial infection criteria. We assessed the incidence, clinical and laboratory findings, diagnostic procedures and surgical approach in these cases. RESULTS: The incidence of intracystic bacterial infection was steady throughout the study period. Clinical and laboratory data are non-specific. Diagnostic procedures for intracystic bacterial infection, including liver computed tomography scan, are of limited value. Although both, radical and non radical surgical approaches, were used in these patients, mortality was associated with non-radical surgery. The morbidity rate was high regardless of the procedure used. CONCLUSIONS: Our current goal in the management of intracystic bacterial infection, if overall condition of the patient is good, is to carry out a total or subtotal cystectomy in order to avoid mortality and hydatid disease relapses. PMID- 11020878 TI - Subdivision of liver anterior segment into two units according to hepatic venous drainage. AB - BACKGROUND/AIMS: In anatomic liver resection, consideration of the distribution of the hepatic vein is important for maximizing remnant liver function. We have examined the anterior segment of the liver tomographically and subdivided it according to the hepatic venous distribution. METHODOLOGY: Thirty patients in whom liver tumors were diagnosed and who were examined by three-dimensional computed tomography were reviewed. Portal and hepatic venous distributions in the anterior segment were analyzed using the tomograms obtained, and the anterior segment was divided into the ventral and dorsal units. RESULTS: S8d was present in only 23 cases, out of which in 14 cases, drainage veins were present. When P8d was supplied by P8, S8d together with S8a were classified into the ventral unit (8 cases). When P8d was supplied by P8c, S8d and S8b and S8c were in the dorsal unit (4 cases). S5a was in the ventral unit in 26 cases and the dorsal unit in 4 cases. S5b was in the ventral unit in 11 cases and in the dorsal unit in 19 cases. The ventral unit was drained by the middle, and the dorsal by the right, hepatic vein. CONCLUSIONS: Three-dimensional computed tomography made it possible to classify the anterior segment into ventral and dorsal units according to the distribution of the hepatic vein. This new classification makes possible new operative procedures in which resection is limited to one of the units with or without one or more segments, and which result in better remnant liver function. PMID- 11020879 TI - Action of somatostatin analogue (octreotide) on experimental hepatic encephalopathy in rats. AB - BACKGROUND/AIMS: We investigated the effect of a somatostatin analogue (octreotide) on hepatic encephalopathy in a rat model. Fulminant hepatic failure was induced by thioacetamide twice daily for 3 consecutive days. METHODOLOGY: Animals with hepatic encephalopathy grade III were divided into 2 groups. The groups received saline (control) or octreotide. In both groups the distance traveled in the open field activity, neurological score and mortality time were evaluated before and after the treatment. RESULTS: In the control group the motor activity was 13.7 +/- 6.4 and 12.9 +/- 5.5 cm/10 min, the neurological score was 8.4 +/- 0.9 and 8.5 +/- 1.3 before and after the treatment, respectively. In the octreotide group the motor activity was 11.4 +/- 5.0 and 10.4 +/- 3.5 cm/10 min, the neurological score was 8.8 +/- 1.5 and 8.6 +/- 0.9 before and after the treatment, respectively. Mortality times in the saline and octreotide group were 76.1 +/- 28.1 and 89.7 +/- 46.5 min, respectively. All parameters of this study were statistically not significant. CONCLUSIONS: This study demonstrated that somatostatin analogue, octreotide does not effect hepatic encephalopathy in an experimental rat model. PMID- 11020880 TI - Small hepatocellular carcinoma with minute satellite nodules. AB - BACKGROUND/AIMS: To investigate the clinicopathologic characteristics of small hepatocellular carcinoma with minute satellite nodules. METHODOLOGY: We investigated the clinicopathologic characteristics of 131 solitary small (< or = 2.0 cm in diameter) hepatocellular carcinomas including 105 hepatocellular carcinomas without minute satellite nodules and 17 hepatocellular carcinomas with minute satellite nodules smaller than 5 mm, and also discuss the clinical significance. RESULTS: None of the clinical backgrounds of the patients and pathologic features of the main tumor, except for the average of preoperative serum alpha-fetoprotein, were significantly different between the two groups. Firstly, minute satellite nodules demonstrated that the maximum diameter of all minute satellite nodules was 1.5-4.0 mm, secondly, the moderately to poorly differentiated hepatocellular carcinomas had 4 or more minute satellite nodules within 1 cm from the main tumor, while well differentiated hepatocellular carcinomas may have 1 or 2 minute satellite nodules 6 cm or more away, and thirdly, 4 or more minute satellite nodules may present within 1 cm in intrahepatic metastasis cases, while 1 or 2 minute satellite nodules may be present 6 cm or more away from the main tumor in multicentric occurrence cases. CONCLUSIONS: At least 13% of solitary small hepatocellular carcinomas had preoperatively undetectable minute satellite nodules. In case of moderately to poorly differentiated hepatocellular carcinomas, hepatic resection as well as percutaneous ethanol injection should be performed including the surrounding liver tissue at least 1.0 cm from the main nodule. On the other hand, in well differentiated hepatocellular carcinomas, which may indicate multicentric occurrence, closer observation and careful follow-up after therapy are recommended. PMID- 11020881 TI - Coagulation and cytokine profile after major hepatobiliary surgery in Chinese patients. AB - BACKGROUND/AIMS: Hypercoagulability and increased circulating cytokine levels have been separately reported after surgical operations. We investigated whether the cytokine level and postoperative coagulation profile would change similarly in Chinese patients undergoing major hepatobiliary surgery. METHODOLOGY: Serial serum levels of IL-6, IL-1 beta and TNF-alpha were measured by colorimetric ELISA. Serial coagulation profile was measured by thrombelastography. Hemoglobin and white cell counts were also measured serially. RESULTS: Only mild hypercoagulability was found at the 12th and 144th hours, evidenced by shortening of r-time and k-time on thrombelastography (P < 0.05). IL-6 and IL-1 beta levels were significantly raised from the 12th hour (P < 0.01) while TNF-alpha level was unchanged. From the 3rd hour to the 48th hour, there was negative correlation between IL-6 levels and various thrombelastographic parameters such as coagulation index (r = -0.777, P < 0.001); k-time (r = 0.478, P < 0.05); maximum amplitude (r = -0.688, P < 0.01) and angle (r = -0.665, P < 0.01). CONCLUSIONS: We observed a much milder degree of hypercoagulability in Chinese patients after major hepatobiliary surgery as compared to those reported in other operations in non-Chinese patients. Unlike in sepsis where a close link between increased cytokine levels and hypercoagulability has been reported, increase in IL-6 was associated with less hypercoagulability in Chinese patients after major hepatobiliary surgery. PMID- 11020882 TI - Interferon-alpha treatment of children with chronic hepatitis D virus infection: the Greek experience. AB - BACKGROUND/AIMS: The therapeutic experience of interferon-alpha therapy against hepatitis D virus infection in affected children is rather limited. For this reason, we conducted a retrospective study (duration: 1991-1995) in order to evaluate the efficacy and the safety of interferon-alpha in children suffering from chronic hepatitis D in Northwestern Greece. METHODOLOGY: Seven children who were found to be infected with HDV in a total of 324 children seropositive for hepatitis B virus infection during the 5-year period of the study were treated with interferon-alpha, 3 x 10(6) U/m2 body surface area, intramuscularly or subcutaneously, 3 times weekly for 1 year (after an informed consent obtained from their parents). Patients were assessed monthly by hematological serological and biochemical tests. Clinical progress, levels of serum alanine aminotransferase, hepatitis D ribonucleic acid (HDV-RNA) and hepatitis B deoxyribonucleic acid (HBV-DNA), seroconversion of hepatitis B surface antigen (HBsAg) and Hepatitis Be Antigen (HBeAg) and liver histology were used as response criteria. RESULTS: Posttreatment alanine transferase levels were significantly reduced (P < 0.05) but Immunoglobulin M and total anti-hepatitis D virus (anti-HDV) antibodies remained positive in all, while hepatitis D ribonucleic acid persisted positive in 4 cases. In addition, no seroconversion of HBsAg or HBeAg was noted and the liver histology progress was disappointing. Side effects including mild fever, arthralgias and malaise and reversible neutropenia and thrombocytopenia were common, but not particularly disturbing. Nevertheless, the children remained fully active on treatment, felt well and attended school. Initially 4 children had been below the 10th percentile for weight and height. All thrived during treatment and two crossed above the 10th percentile indicating height velocity and body mass index increase. CONCLUSIONS: The administration of regular interferon-alpha doses for treating children with chronic hepatitis D was safe as attested by the mild side effects and the objective clinical criteria regarding their growth, but relatively ineffective. Although the prevalence of hepatitis D virus infection is now generally decreased, this study indirectly indicates that more effective agents and new approaches at the molecular level of the hepatitis D virus genome are urgently warranted for its control in individuals already infected with the virus. Finally, the poor therapeutic results in the present study further enhance the necessity of the expanded vaccination against Hepatitis B virus according to the World Health Organization's recommendations. PMID- 11020883 TI - Combination of preoperative embolization of the right portal vein and hepatic artery prior to major hepatectomy in high-risk patients: a preliminary report. AB - BACKGROUND/AIMS: Preoperative transhepatic portal vein embolization may not always be sufficient to achieve the desired changes in contralateral hepatic volume and function. The beneficial role of additional transcatheter arterial embolization performed after inadequate response to preoperative transhepatic portal vein embolization is described. METHODOLOGY: Four patients underwent both preoperative transhepatic portal vein embolization and transcatheter arterial embolization, and 6 control patients underwent preoperative transhepatic portal vein embolization only. Changes in right liver lobe volume fraction, residual left lobe volume fraction, and prediction score (low-risk, < 45; borderline, 45 55; high-risk > 55); were evaluated. RESULTS: 1) The change in right liver lobe volume after both preoperative transhepatic portal vein embolization and transcatheter arterial embolization (volume after/before) was 0.75 times that of the original level whereas after preoperative transhepatic portal vein embolization, they were only 0.81 times that of the original level. 2) The change in residual left liver volume after both preoperative transhepatic portal vein embolization and transcatheter arterial embolization (volume after/before) was 1.40 times that of the original level whereas after preoperative transhepatic portal vein embolization they were only 1.30 times than the original level. The changes in left liver volume after preoperative transhepatic portal vein embolization/transcatheter arterial embolization was more favorable than those after preoperative transhepatic portal vein embolization only. 3) The change in prediction score after both preoperative transhepatic portal vein embolization and transcatheter arterial embolization (after/before) was 0.81 times that of the original level. All prediction score in high-risk patients recovered to the borderline or safety zone. Change after preoperative transhepatic portal vein embolization only (before/after) was 0.87 times that of the original level. 4) All 4 patients who underwent both preoperative transhepatic portal vein embolization and transcatheter arterial embolization received right hepatic lobectomy successfully and returned to their normal life style. CONCLUSIONS: Preoperative occlusion of right hepatic inflow vessels increased the volume and function of the contralateral lobe where high-risk patients recovered to the borderline zone for major hepatic resection. PMID- 11020884 TI - Correlation between plasma and hepatic phosphatidylcholine hydroperoxide, energy charge, and total glutathione content in ischemia reperfusion injury of rat liver. AB - BACKGROUND/AIMS: Oxygen-derived free radicals are believed to be responsible for the hepatocellular injury leading to liver failure following ischemia-reperfusion in liver, endotoxemia and many other life-threatening illnesses. This study was designed to investigate the reactive oxygen species interaction in lipid peroxidation, the adenosine and energy charge levels of liver cells, and total glutathione content in ischemic-reperfusion injury of liver in rat. METHODOLOGY: To prevent intestinal congestion during the clamping of vascular structures, subcutaneous transposition of the spleen was done beforehand. Four to six weeks later, after the development of natural portal-systemic shunts, occlusion of the portal vein, hepatic artery and bile duct was performed for different periods; blood and liver samples were taken at different intervals after the release. On the basis of the ischemia-reperfusion time, the rats were divided into the following 9 groups: 30/0, 30/30, 30/60, 60/0, 60/30, 60/60, 90/0, 90/30, and 90/60. The following parameters were measured: total hepatic glutathione content, adenosine values (ATP, ADP, AMP), energy charge, phosphatidylcholine hydroperoxide (PCOOH) concentrations in liver and plasma, and serum transaminases (AST, ALT). Decreased liver glutathione stores (an indicator of increased oxidative stress), increased serum hepatic transaminases (an indicator of hepatocellular injury), and increased PCOOH (an indicator of cellular-membrane lipid peroxidation) were noted. RESULTS: The ATP level and energy charge diminished significantly with the increase in duration of ischemia and reperfusion. A close correlation between the PCOOH levels in plasma and liver was observed. Extreme damage was noted in the 90-minute ischemia with 60-minute reperfusion group. The hepatic total glutathione level was reduced to the lowest level in the 90/60 group and it correlated with the energy charge level, denoting the highest degree of oxidative stress sustained by the liver cells in this group. CONCLUSIONS: These results indicated that prolonged hepatic ischemia with reperfusion produced bursts of oxygen-derived free radicals which overwhelmed the defense mechanisms of the cells, with a resultant decrease in energy charge associated with an increase in membrane lipid peroxidation. These findings not only provide confirmation of previously reported hepatocellular injury by free radicals generated after reperfusion, but they also establish the use of PCOOH analysis in liver and plasma as a sensitive and specific indicator of the injury process in time. The plasma PCOOH level may be a useful indicator of free radical induced hepatic membrane lipid peroxidation during ischemia-reperfusion, and might be employed in clinical studies of the therapeutic effects of drugs in various liver diseases, as well as for determining the prognosis after different kinds of hepatic operations. PMID- 11020885 TI - Hepatic resections for non-colorectal metastases: forty resections in 35 patients. AB - BACKGROUND/AIMS: Hepatic resection, though now an accepted practice for colorectal primary tumors, is poorly documented for non-colorectal metastases. However, the few series reported suggest that this approach may lead to a significant increase in survival. METHODOLOGY: Study of 40 cases of resection in 35 patients with non-colorectal hepatic metastasis to define the role of hepatic resection between 1986 and 1997. RESULTS: Resection was performed for 5 metastases of ovarian and fallopian tube carcinoma, 8 gastrointestinal tract adenocarcinomas, 8 endocrine tumors, 8 sarcomas and 6 miscellaneous metastases, involving 17 lobectomies, 3 trisegmentectomies, 5 lateral segmentectomies and 15 non-anatomical local resections. Survival at 1, 2 and 5 years was 54 +/- 8, 42 +/ 8 and 27 +/- 8%, respectively. Hepatic metastases of gastrointestinal tract adenocarcinomas were found to have the poorest prognosis (median time: 13 months), and genital tract adenocarcinomas the best (27 months). CONCLUSIONS: Some carefully selected patients may benefit from liver resection for non colorectal metastases. PMID- 11020886 TI - Surgical indications for advanced hepatocellular carcinoma. AB - BACKGROUND/AIMS: The aim of this study is to clarify the limitations of hepatectomy for advanced hepatocellular carcinoma. METHODOLOGY: Fifty-six patients with Stage 4 hepatocellular carcinoma were retrospectively studied, and the prognostic factors were both univariately and multivariately analyzed. The VI score, which was defined as the degree of portal vein invasion (VP) multiplied by the degree of intrahepatic metastases (IM), was introduced as a new prognostic indicator. RESULTS: A univariate analysis revealed the following significant variables: hypertension, esophageal varices, Child's classification B or C, a bilirubin value of over 1.0 mg/dL, a albumin value of below 3.5 g/dL, a GOT value of over 100 IU/L, an AFP value of over 1000 ng/mL, a history of tumor rupture, Stage 4B, a tumor size of over 5 cm, VP3, IM3, and the VI score of no less than 6. A multivariate analysis demonstrated the following 4 variables to be independent prognostic indicators: a Stage of 4B, a VI score of no less than 6, a Child's classification of B or C, and a tumor size of over 5 cm. Furthermore, no long-term survivors were found in patients with either Stage 4B HCC or a hepatocellular carcinoma having a VI score of more than 6. At the present time, either Stage 4B or a hepatocellular carcinoma having a VI score > or = 6 are considered to be factors which means the limitation of hepatectomy alone. Furthermore, an advanced hepatocellular carcinoma with either poor liver function or a hepatocellular carcinoma with a size of over 5 cm should be carefully evaluated before determining its appropriateness for hepatectomy. CONCLUSIONS: The VI score is therefore suggested to be a useful prognostic indicator for determining the surgical indications for advanced hepatocellular carcinomas. PMID- 11020887 TI - Resected cases of hepatocellular carcinoma detected after interferon therapy for chronic hepatitis C. AB - BACKGROUND/AIMS: Interferon therapy decreases the incidence of hepatocellular carcinoma in patients infected with hepatitis C virus. However, hepatocellular carcinoma was detected after interferon therapy in some patients. METHODOLOGY: Of the 167 patients who underwent liver resection for hepatitis C virus-related hepatocellular carcinoma between 1993 and September 1998, the carcinoma was detected after interferon therapy in 11 patients. The clinicopathologic findings in these 11 patients were studied. RESULTS: The response to interferon was complete (n = 4), partial (n = 4), or no response (n = 3). Hepatocellular carcinoma was detected 2 months to 3 years 9 months, after interferon therapy. The interval period from the end of interferon therapy to the detection of the carcinoma were significantly correlated with the longest diameter of the main tumor (P = 0.0043), indicating that most carcinomas have already developed before the end of interferon therapy. In one non-responder, multicentric carcinogenesis occurred after liver resection for primary hepatocellular carcinoma. Another patient with advanced hepatocellular carcinoma died of the recurrence. CONCLUSIONS: Surveillance for hepatocellular carcinoma must be performed even in patients successfully treated with interferon because occult carcinoma may have developed before or during the therapy. PMID- 11020888 TI - Doubly-armed short sutures are useful for microsurgical hepatic artery reconstruction in living-related liver transplantation. AB - Although microsurgical hepatic artery reconstruction has become the essential technique in pediatric and living-related liver transplantation, it remains a technical challenge. We describe here our microsurgical hepatic artery reconstruction technique in living-related liver transplantation using a doubly armed short suture. From August 1998 to March 1999, 8 patients underwent living related liver transplantation at the Kyushu University. For these 8 patients, microsurgical reconstruction with doubly-armed short sutures were undertaken. After preparing the operative field for microsurgery, operative microscopy with a continuous zoom magnification is introduced. Interrupted sutures using doubly armed short sutures are performed in a biangular stay-sutured method with a double clip. None of the 8 patients developed hepatic artery thrombosis. We conclude that the use of doubly-armed sutures allows safe and reliable in microsurgical hepatic artery reconstruction in living related liver transplantation. PMID- 11020889 TI - Conversion of unresectable hepatoma to resectable. Report of a case and review of the literature. AB - One of the most important reasons that hepatocellular carcinoma displays a poor prognosis, is the low resectability rate at the time of the diagnosis. In this study, we report a case of unresectable hepatocellular carcinoma converted to resectable after transcatheter arterial chemoembolization. In addition a review of the literature is attempted. PMID- 11020890 TI - Liver perforation: a serious complication of percutaneous acetic acid injection for hepatocellular carcinoma. AB - A 75-year-old cirrhotic man, after undergoing percutaneous acetic acid injection therapy for hepatocellular carcinoma, presented high fever and right hypochondralgia. Computed tomography disclosed an area of low attenuation in the liver and a crescent-shaped intraperitoneal lesion adjacent to it. We diagnosed liver perforation and localized peritonitis due to the leakage of acetic acid following acetic acid injection. It is important to consider this serious complication when evaluating the indications for percutaneous acetic acid injection for hepatocellular carcinoma. PMID- 11020891 TI - Portal-systemic encephalopathy due to a congenital extrahepatic portosystemic shunt: three cases and literature review. AB - Extrahepatic portal-systemic encephalopathy due to congenital extrahepatic portosystemic shunt has so far been rarely reported in the literature. We herein report 3 such cases without liver cirrhosis or portal hypertension which were presented with the chief complaint being disturbance of consciousness and abnormal behavior. In all cases the brain computed tomography scan revealed no pathological findings, while electroencephalogram showed a diffuse slow activity with triphasic waves. The laboratory data revealed a high serum ammonia level. Percutaneous transhepatic portography demonstrated portosystemic shunts. After these shunts were surgically occluded, the serum ammonia level reached a normal range and encephalopathy disappeared. A liver biopsy also revealed neither fibrosis nor cirrhosis in any of the cases. The 23 previously reported cases are also discussed. PMID- 11020893 TI - Hepatocellular carcinoma responding to chemotherapy with 5-FU. AB - A 69-year-old man with unresectable hepatocellular carcinoma and portal vein tumor thrombus was treated by chemotherapy with 5-fluorouracil. A dose of 500 mg/day of 5-fluorouracil was continuously administered via a central venous catheter. After 4 months, the alpha-fetoprotein level was decreased from 50,000 ng/mL to 4,760 ng/mL. Computed tomography revealed disappearance of the low density area in the liver parenchyma, but the portal vein tumor thrombus was not changed. After 6 months, pancytopenia appeared and continuous infusion of 5 fluorouracil was stopped. After 8 months, the patient died of pneumonia, at which time the alpha-fetoprotein level was 12,000 ng/mL. Continuous intravenous infusion of 5-Fluorouracil was effective against unresectable primary hepatocellular carcinoma, but had little influence on portal vein tumor thrombus. PMID- 11020892 TI - Recurrent hepatocellular carcinoma presenting with superior vena cava syndrome. AB - A 45-year-old male received wedge resection for his small hepatocellular carcinoma in April 1989 and extended right lobectomy for tumor recurrence 8 months later. Unfortunately, recurrent hepatic tumor with lung metastases were found 18 months after the second operation. Both the hepatic and pulmonary recurrent tumors were resected and transcatheter arterial embolization was added for the residual hepatic tumors. He remained symptom free for another 18 months. However, mediastinal lymphadenopathy, superior vena cava thrombus with superior vena cava syndrome, cardiac and brain metastases developed subsequently. He died of increased intracranial pressure. It is rare for hepatocellular carcinoma to have mediastinal metastases, superior vena cava thrombus and superior vena cava syndrome. PMID- 11020894 TI - Spontaneous total necrosis of hepatocellular carcinoma: report of a case. AB - We report on a 50-year-old man with a liver mass that, when surgically resected, was found to be a hepatocellular carcinoma that had undergone spontaneous complete necrosis without previous treatment. Histologically, no viable tumor cells were observed. The postoperative course was uneventful, and the patient is alive without evidence of recurrence about 5 years after surgery. Spontaneous total necrosis of hepatocellular carcinoma without previous treatment is rare. In particular, spontaneous total necrosis of a hepatocellular carcinoma that has been proven by histologic examination of a surgically resected liver specimen is extremely rare; only three cases of spontaneous complete necrosis of hepatocellular carcinoma have been reported previously. In this report, we present our unusual case and discuss possible causes of spontaneous total necrosis or regression of hepatocellular carcinoma. PMID- 11020895 TI - Combined hepatocellular-cholangiocarcinoma presented with massive pulmonary embolism. AB - A 30-year-old HBsAg-positive woman was admitted to the hospital because of 6 days of progressive shortness of breath. She was in severe respiratory distress with circulatory collapse. She had an enlarged liver but no stigmata of chronic liver disease or signs of cirrhosis. She had rapidly developed respiratory arrest and was transferred to intensive care unit. Heart ultrasonography and Doppler scan showed right heart straining and high pulmonary artery pressure. Despite cardiovascular and respiratory support she died a few hours after admission. Autopsy revealed combined hepatocellular-cholangiocarcinoma infiltrating the entire liver, metastatic invasion of lung blood vessels and absence of right ventricular hypertrophy. The incidence of hepatocellular-cholangiocarcinoma, a variant of hepatocellular carcinoma, is roughly 2-3% and the presenting symptoms are abdominal pain, weight loss, jaundice, fever or decompensation of liver disease. Associated HBsAg positivity and cirrhosis are reported in 20-30% and 60% of patients, respectively. Metastases to lungs are relatively frequent but this is the first report of hepatocellular-cholangiocarcinoma presented with acute respiratory distress due to massive pulmonary embolism. PMID- 11020897 TI - Preoperative staging of periampullar cancer with US, CT, EUS and CA 19-9. AB - BACKGROUND/AIMS: Several diagnostic methods are available for preoperative staging of periampullary cancers. It happens that on the basis of preoperative findings the patient is prepared for radical surgery, but during the operation the lesion turns out to be unresectable. METHODOLOGY: We studied 43 patients operated on for periampullary cancer. Preoperatively, all patients were evaluated by ultrasonography, computed tomography, endoscopic ultrasonography and the level of carbohydrate antigen 19-9 was assessed. Statistical parameters were calculated and compared. RESULTS: Endoscopic ultrasonography has specificity of 70.8% and positive predictive value regarding tumor resectability 55.8%. Computed tomography has a specificity of 45.8% and carbohydrate antigen 19-9 of 66.7%. Positive predictive value for computed tomography and carbohydrate antigen 19-9 is 40.6% and 52.6%, respectively. Ultrasonography is the least accurate method with specificity of 12.5% and positive predictive value 29.7%. A combined use of different diagnostic methods has higher positive predictive value, highest (65.1%) being found for the combination of endoscopic ultrasonography and carbohydrate antigen 19-9. Statistical tests showed statistically significant differences between diagnostic methods. CONCLUSIONS: Among the diagnostic methods studied, endoscopic ultrasonography showed the highest accuracy in predicting tumor resectability. The use of either endoscopic ultrasonography and carbohydrate antigen 19-9 or computed tomography and carbohydrate antigen 19-9 is accurate enough for assessing tumor resectability. PMID- 11020896 TI - The prognosis of intraductal papillary mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: Intraductal papillary mucinous tumors of the pancreas have been recognized as a distinct clinical entity. However, their biological behavior has not been clearly defined. The aim of this study was to examine the prognosis of this tumor, to clarify the biological behavior and determine the most appropriate treatment. METHODOLOGY: Correlations between prognosis of operated cases and histopathologic features were investigated. RESULTS: In 105 patients with characteristic clinical features of intraductal papillary mucinous tumors, the lesions were classified as hyperplasias in 21%, intraductal tumors in 48% and invasive carcinomas in 31%. Minimal invasion was apparent in 25%, lymph node metastasis in 21%, and fistula formation in 31% of the invasive lesions. Non invasive and minimally invasive intraductal papillary mucinous tumors were essentially free from risk of tumor recurrence. Other invasive intraductal papillary mucinous tumors showed a significantly poor prognosis. CONCLUSIONS: Because of the variation in pathological characteristics, patient outcome and the possibility of differential diagnosis, the treatment might be recommended as follows: the case of hyperplasia can be followed-up with close surveillance. Non invasive and minimally invasive intraductal papillary mucinous tumors should be operated with function-preserving minimal pancreatectomy. For patients with invasive intraductal papillary mucinous tumors evident with preoperative imaging modalities, radical operations with lymph node dissection might be needed. PMID- 11020898 TI - Pancreatogastrostomy (PG) after pancreatoduodenectomy with or without duct-to mucosa anastomosis for the small pancreatic duct: short- and long-term results. AB - BACKGROUND/AIMS: Pancreatogastrostomy, generally considered to pose less postoperative complications for a small-duct pancreas after pancreatoduodenectomy than pancreatojejunostomy, has usually been conducted with an invagination method, which can cause obstruction of the duct during the follow-up. The purpose of this study was to investigate the short- and long-term results with special reference to the patency of the main pancreatic duct after pancreatogastrostomy performed on a small-duct pancreas, combining an invagination technique with a duct-to-mucosa approximation. METHODOLOGY: Out of 73 patients with a nondilated pancreatic duct, 24 in an earlier series underwent pancreatogastrostomy only with an invagination. The other 49 in a later period had an additional duct-to-mucosa anastomosis. RESULTS: Four patients (5.5%) developed a minor anastomotic leak of the pancreas which healed uneventfully within 16-41 days without mortality. Long term results revealed that the pancreatic duct tended to dilate after pancreatogastrostomy without mucosal adaptation, while the new method of pancreatogastrostomy with duct-to-mucosa anastomosis left the diameter of the pancreatic duct unchanged. Body weight and peripheral blood glycohemoglobin A1c retained preoperative levels, irrespective of the mucosal anastomosis. CONCLUSIONS: Pancreatogastrostomy with mucosa-to-mucosa anastomosis appears to be a useful method of pancreatic reconstruction in both the short- and long-term. PMID- 11020899 TI - Intraoperative and conformal external-beam radiation therapy in patients with locally advanced pancreatic carcinoma; results from a feasibility phase II study. AB - BACKGROUND/AIMS: Chemoradiation therapy is widely indicated to patients with locally advanced pancreatic carcinoma, though the capability of radiotherapy alone is not assessed enough. The purpose of this study is to clarify the efficacy and safety of a more intensive radiotherapy for those patients. METHODOLOGY: Fifteen patients were enrolled in a feasible phase II study of treatment with intraoperative radiation therapy (25 Gy), followed by conformal external-beam radiation therapy (40 Gy in 20 fractions, 5 times/week). The antitumor effect was evaluated on the early phase of dynamic computed tomography image. RESULTS: The full irradiation dose was feasible in 12 of 15 patients. The overall response rate was 40% (1 complete and 5 partial responses). Grade 3 toxicity was observed in 2 patients (13%) with nausea/vomiting or anorexia. One patient developed gastric ulcer and died of gastrointestinal bleeding 12 months after intraoperative radiation therapy. The median survival time was 11.1 months, and the 2-year survival rates were 13%. Survival for more than 2 years was observed in 2 of the 6 responders. CONCLUSIONS: The above radiotherapy is considered to be active for the locally advanced pancreatic cancer with acceptable toxicity, when the gastrointestinal tract is excluded from the radiation field. This should be further assessed in late phase II studies involving a large number of patients. PMID- 11020900 TI - Prospective and randomized study of gabexate mesilate for the treatment of severe acute pancreatitis with organ dysfunction. AB - BACKGROUND/AIMS: Exaggerated production of various proteases may account for the late presence of organ dysfunction in acute pancreatitis. This study examined the effects of early administration of gabexate mesilate on the condition of patients with severe acute pancreatitis and organ dysfunctions. METHODOLOGY: Fifty-two patients with acute pancreatitis and organ dysfunction were enrolled. The treatment group included 26 patients receiving intravenous gabexate mesilate infusion at a dose of 100 mg/hr for 7 days. APACHE-II score, clinical and biochemical parameters were monitored intensively. RESULTS: Coagulopathy ileus, and abdominal pain was significantly improved with gabexate mesilate. Gabexate mesilate reduced the necessity for surgical intervention and peritoneal lavage. The 7-day-mortality and 90-day-mortality rates were also significantly reduced with gabexate mesilate therapy. CONCLUSIONS: There are strong indications from this study that early intravenous gabexate mesilate infusion results in improved survival in acute pancreatitis with organ dysfunctions. PMID- 11020901 TI - Intraoperative radiotherapy and bypass surgery for unresectable pancreatic cancer. AB - BACKGROUND/AIMS: Conflicting results have been reported concerning the usefulness of radiotherapy for unresectable pancreatic cancer. We evaluated the clinical efficacy of intraoperative radiotherapy and/or external beam radiotherapy in combination with bypass surgery. METHODOLOGY: Twenty-six patients with unresectable pancreatic cancer (16 in Stage II-III and 10 in Stage IV) were treated with intraoperative radiotherapy plus external beam radiotherapy (16 patients) or intraoperative radiotherapy alone (10 patients). The dose of intraoperative radiotherapy was either 25 or 30 Gy and the external beam radiotherapy dose was 31-60 Gy. The feasibility and clinical outcome were analyzed. RESULTS: The median survival time for Stage II-III and Stage IV were 11.5 and 6.5 months, respectively. The difference between Stage II-III and Stage IV in survival patterns was statistically significant (P < 0.05). For Stage II III patients, the survival curves between the groups of intraoperative radiotherapy plus external beam radiotherapy and intraoperative radiotherapy alone were not significantly different, and only performance status was a significant factor in the prognosis (P < 0.05). Gastrointestinal bleeding was noted in 8%, but did not occur in the patients treated with an external beam radiotherapy dose less than 50 Gy. Palliative radiation was successfully performed to relieve pain, jaundice and appetite-loss and to shorten the hospital stay. CONCLUSIONS: The combination therapy with intraoperative radiotherapy and bypass surgery is considered to be tolerable and effective for unresectable pancreatic cancer, and also may improve the quality of life of the patients. PMID- 11020902 TI - Complete drainage of pancreatic juice from the cut edge of the pancreas after subtotal left-side pancreatectomy. AB - The management of the cut stump after a subtotal left-side pancreatectomy is sometimes difficult compared with that after distal pancreatic resection, which is performed at or to the left of the superior mesenteric vein. This report describes a simple and effective drainage method of leaking pancreatic juice from the cut stump after subtotal left-side pancreatectomy for pancreatic adenocarcinoma located in the body and tail. Since July 20, 1994, we have applied this method to 12 patients with carcinoma of the body and tail of the pancreas other than the present case and as yet have never experienced serious postoperative complications. PMID- 11020903 TI - Pancreatic adenocarcinoma in a patient with Peutz-Jeghers syndrome: report of a case and literature review. AB - We present a rare case of pancreatic adenocarcinoma in a 47-year-old man with the Peutz-Jeghers syndrome. The patient underwent pancreatoduodenectomy with partial resection of the portal vein. We also review the current literature concerning Peutz-Jeghers syndrome associated with malignant tumors, especially pancreatic cancer. To our knowledge, this is the first report of a Peutz-Jeghers syndrome patient with pancreatic cancer having pancreatoduodenectomy and pathologically diagnosed with invasive ductal adenocarcinoma of the pancreas. The Peutz-Jeghers syndrome patients with pancreatic cancer were relatively young. As the pancreatic cancer in these patients was advanced and most were unresectable at diagnosis, the prognoses of these patients were extremely poor. Surgical resection offers the only chance for cure or long-term survival for Peutz-Jeghers syndrome patients, if the tumor is localized without distant metastasis. Therefore, screening even for young patients with Peutz-Jeghers syndrome is necessary for early detection of cancer. PMID- 11020904 TI - Common bile duct and pancreatic injury after extracorporeal shock wave lithotripsy for renal stone. AB - Common bile duct and pancreatic injury are rare complications following extracorporeal shock wave lithotripsy. We reported a case of peripancreatic abscess with inflammation change of common bile duct and pancreatic head following extracorporeal shock wave lithotripsy for right renal stone. Its anatomical location and subsequent clinical course suggest it was related to trauma caused by the shock wave. It should be considered a relative contraindication to apply extracorporeal shock wave lithotripsy in cases of right renal stone associated with large gall stones. PMID- 11020905 TI - Biliopancreatic fistula associated with intraductal papillary-mucinous pancreatic cancer: institutional experience and review of the literature. AB - Intraductal papillary-mucinous tumour is clinicopathologically characterized by papillary growth and mucin production within the pancreatic duct system. The category includes a wide range of dysplasia, ranging from adenoma to carcinoma, the latter designated as intraductal papillary-mucinous cancer. In general, the tumor renders a favorable prognosis after complete resection. However, intraductal papillary-mucinous tumor with overt invasion outside the gland has been reported to have a poor prognosis, as is the case with the usual type of duct cell cancer of the pancreas. We experienced two cases of intraductal papillary-mucinous cancer with obstructive jaundice due to impaction of thick mucus protruding from the pancreas via a "spontaneous" biliopancreatic fistula. Preoperative examinations of both patients showed a large intraductal papillary mucinous tumor in the head of the pancreas with fistula formation between the intrapancreatic portion of the common bile duct and the main pancreatic duct. Histopathological investigation of the two resected specimens suggested that the fistula may not have developed from invasion by papillary or tubular adenocarcinoma, but from compression and destruction of the intercalating tissues by abundant mucinous secretion. The first patient died of peritoneal carcinomatosis with clinicopathologic features of pseudomyxoma peritonei 6 years after surgery. The second patient is alive and has been well for 2 years postoperatively. Review of the world literature showed that half of the patients with intraductal papillary-mucinous cancer plus biliopancreatic fistula had no stromal invasion around the fistula, indicating that the fistula might have been caused by mechanical pressure. However, the other half of the cases did have stromal invasion around the fistula. Two-thirds of these cases, including our own patients, had foci of mucinous carcinoma in the stroma around the fistulization, implying that mucinous lakes in the stroma may have served as part of the "waterway" from the pancreatic duct to the bile duct, assisted by increased pressure by mucus production. Since intraductal papillary-mucinous cancer with biliopancreatic fistula has a comparatively favorable prognosis, surgical resection should be considered. PMID- 11020906 TI - Resection of a non-functioning islet cell carcinoma occupying the entire pancreas. AB - A 46-year-old woman with jaundice was found to have enlargement of the entire pancreas on computed tomography. Endoscopy revealed a tumor at the papilla of Vater, although examination of biopsy specimens did not demonstrate a malignancy. The jaundice resolved spontaneously, but reappeared at 7 months. Also noted was increased swelling of the papilla. The serum pancreatic endocrine and exocrine hormone concentrations were within the reference ranges. At surgery, a hard mass was found occupying the entire pancreas. Examination of incisional biopsy specimens from the pancreatic mass and papilla of Vater revealed an islet cell tumor. A total pancreatectomy was performed. The tumor extended from the pancreatic tail to the papilla of Vater. Histopathologic and immunopathologic examination revealed a non-functional islet cell carcinoma. There was no evidence of recurrence at 3 years. A good outcome can be achieved with resection of non functioning islet cell carcinomas, even if they are advanced. PMID- 11020907 TI - Pancreatic cancer: current factors and options to determine future research. AB - There are many factors to consider when reviewing the fact that the prognosis for patients diagnosed with pancreatic carcinoma is "dismal." For this reason, we reviewed various articles regarding the diagnostic factors, treatment and prognostic factors including resection and laparoscopy, along with modalities of treatment to establish the objectives for future developments. We believe that as our population ages, we will see fundamental changes in the factors, options and modalities of treatment for this lethal disease. Public health efforts which are moving for the screening for early cancer detection, a better understanding of the tumor biology and efforts to reduce the gap between the clinical practice and the scientific understanding may ultimately reduce the burden of cancer in society. PMID- 11020908 TI - Six-day therapy with ranitidine bismuth citrate plus low-dose clarithromycin and tinidazole to cure Helicobacter pylori infection. AB - BACKGROUND/AIMS: Short-term ranitidine bismuth citrate-based triple regimens have been shown to be effective for the eradication of H. pylori. We investigated the efficacy of an eradicating therapy including ranitidine bismuth citrate, low-dose clarithromycin and tinidazole, administered for only 6 days. METHODOLOGY: Forty five consecutive patients, who underwent gastroscopy for symptoms and were found to be H. pylori-positive, were recruited. They received ranitidine bismuth citrate 400 mg b.i.d. plus clarithromycin 250 mg b.i.d. plus tinidazole 500 mg b.i.d., given for 6 days. The medications given in twice daily doses were taken after meals with an interval of 12 h. The H. pylori status was evaluated by means of histology and rapid urease test on admission, and by 13C-urea breath test alone 8 weeks after treatment. RESULTS: All 45 enrolled patients completed the study. Thirty-nine of 45 patients returned H. pylori-negative (both per protocol and intention-to-treat analysis = 87%; 95% confidence interval = from 73-95%), while 6 of 45 were still H. pylori-positive (13%). Slight or mild side effects occurred in 5/45 patients (11%). CONCLUSIONS: Ranitidine bismuth citrate-based triple therapy, containing low-dose clarithromycin and tinidazole, given for only 6 days, yielded high eradication rates with modest side effects. Regimens based on ranitidine bismuth citrate plus two antibiotics at low dosages, administered for less than 7 days, constitute highly promising strategies for eradication of H. pylori. PMID- 11020909 TI - Clinicopathological evaluation of T2-gastric cancer among age groups. AB - BACKGROUND/AIMS: Although the recent results of surgical treatments for T2 gastric cancer (defined as: tumor invasion of the muscularis propria or the subserosa) have been comparatively favorable, frequency of lymph node metastasis is high and recurrence often happens. METHODOLOGY: A retrospective study on T2 gastric cancer in 347 patients who underwent curative resection between 1975 and 1995 was performed to address this issue. These 347 patients were divided into 3 groups according to age: group I (72 cases) < 50 years old; group II (202 cases) not < 50 years but < 70 years old; and group III (73 cases) > or = 70 years old. RESULTS: There was an apparent tendency that gastric cancer in aged patients is more likely to become differentiated cancer and spread easily to the liver. CONCLUSIONS: The data suggest that aged patients should be more carefully followed for any hematogenous metastases including liver metastases. Removal of hepatic metastases or regional hepatic infusion chemotherapy may then salvage some patients even elderly patients. PMID- 11020910 TI - Does nimesulide induce gastric mucosal damage? "A double-blind randomized placebo controlled trial". AB - BACKGROUND/AIMS: In this study, it was aimed to examine the effect of nimesulide, a selective inhibitor of cox-2 enzyme, to the gastric mucosa and to correlate its effect with aspirin. METHODOLOGY: This study was planned as double-blind, randomized and placebo-controlled. Mean age of voluntary persons (n = 32) was 42.3 +/- 2.7. Divided into 3 groups of volunteers were given randomized placebo (n = 10), aspirin (n = 10) (500 mg aspirin, Bayer) and nimesulide (n = 12) (100 mg mesulid, Pfizer) with 50 mL of water after 12 hours fasting period at 08.00 am. Gastroduodenoscopy was performed to the volunteers 3 hours after each therapy. RESULTS: Endoscopic scores of groups were; placebo: 0.20 +/- 0.13, aspirin: 2.8 +/- 0.46, nimesulide: 1.41 +/- 0.51. Lesion scores both in the aspirin group when compared with nimesulide and placebo groups (P < 0.00002, < 0.03), and in the nimesulide group when compared with the placebo group (P < 0.01) were significantly high. The positivity of Helicobacter pylori of groups was found; 67% in placebo, 72% in aspirin, 71% in nimesulide and there was no statistically significant difference in the groups. CONCLUSIONS: It was shown that nimesulide causes significantly serious gastric mucosal lesion when compared with placebo. The lesion score of nimesulide was found less than aspirin. According to the findings, nimesulide should be given carefully just as other analgesics due to the probability of causing gastric lesion. PMID- 11020911 TI - Clinicopathologic differences between early gastric remnant cancer and early primary gastric cancer in the upper third of the stomach. AB - BACKGROUND/AIMS: This study was designed to clarify the clinicopathologic characteristics and survival in early gastric remnant cancer and compare with early primary cancer in the upper third of the stomach. METHODOLOGY: Twenty-five patients with early gastric remnant cancer, who underwent resection at Kanagawa Cancer Center and First Department of Surgery, Yokohama City University between 1974 and 1996 were evaluated in this study. Various clinicopathologic characteristics, such as age, sex, symptoms, size of tumor, depth of invasion, lymph node metastasis, cell differentiation, and survival were investigated and early gastric remnant cancer was compared with early primary cancer in the upper third of the stomach. RESULTS: According to the macroscopic type, protruded type such as I or II type accounted for a great majority in early gastric remnant cancer, while II c depressed type was common in early primary cancer in the upper third of the stomach, comprising 64.2% of all cases. Pathological examination disclosed that well-differentiated carcinoma and mucosal carcinoma were more frequently observed in early gastric remnant cancer than in early primary cancer in the upper-third of the stomach. The 5-year survival rate was 83.5% for early primary cancer in the upper-third of the stomach. In contrast, no patients experienced recurrence after operation for early gastric remnant cancer. CONCLUSIONS: From the view point of clinicopathological evaluation, gastric remnant cancer is a special from of gastric cancer. A follow-up program is important in order to detect early gastric remnant cancer. A low incidence of lymph node metastasis suggests that endoscopic mucosal resection of the tumor or limited operation could be performed under strict indication. PMID- 11020913 TI - Treatment of the gastric stump ulcer: an open study with five drugs. AB - BACKGROUND/AIMS: Despite a great progress in the treatment of peptic ulcer disease, the management of gastric stump ulcers still remains to be established. METHODOLOGY: Eighty-one patients with peptic ulcer developed postoperatively in the gastric remnant were treated in an open trial with 5 antiulcer drugs (cimetidine, omeprazole, sucralfate, colloidal bismuth and misoprostol) characterized by different mechanisms of action. The ulcer healing rate was evaluated endoscopically after 2, 4 and 6 weeks. RESULTS: It was found that after 2 weeks the most rapid ulcer healing was in the omeprazole and cimetidine treated groups, 67 and 43% of healing, respectively. Also after 4 weeks the antisecretors were more effective than gastroprotective drugs; ulcer healing rate for omeprazole was 87% and cimetidine 68%, while for sucralfate, colloidal bismuth and misoprostol 50%, 52%, and 33%, respectively. After 6 weeks all drugs represented very close ulcer healing rates. CONCLUSIONS: Both antisecretory and gastroprotective drugs may be useful in the management of stump ulcers, however, to initiate and accelerate the stump ulcer healing omeprazole appears to be the drug of choice. PMID- 11020912 TI - Cyanocobalamin absorption abnormality in alcoholics is improved by oral supplementation with a fermented papaya-derived antioxidant. AB - BACKGROUND/AIMS: Thirty alcoholic patients and 24 teetotaler dyspeptic patients were considered and underwent baseline blood chemical evaluation and the Schilling test. METHODOLOGY: During gastroscopy, biopsy samples were taken to assay: routine histology, malonyldialdehyde, vitamin E and glutathione concentration and for testing vitamin B12-Intrinsic Factor binding. Examinations were repeated after 1-week supplementation with Bionormalizer. RESULTS: Plasma malonyldialdehyde level and lipid hydroperoxides concentration as well as either malonyldialdehyde and xanthine oxidase concentration in the gastric mucosa in alcoholics were significantly higher than in controls and despite unchanged alcohol consumption, significantly decreased after Bionormalizer supplementation. Gastric mucosal glutathione was markedly depressed in alcoholics and partly recovered after Bionormalizer supplementation. Although the alcoholics showed a normal intrinsic factor secretion in the gastric juice, they exhibited a markedly depressed intrinsic factor-cobalamin binding on the "ex vivo" study. Moreover, nearly 23% of them had an abnormal Schilling test. Both these impairments reverted to normal after Bio-normalizer supplementation. CONCLUSIONS: It can be postulated that the antioxidative action played by Bionormalizer, possibly due to its availability substrates for glutathione synthesis as well as to its effects on local oxidative burst from neutrophil, is able to recover a normal cobalamin absorption. PMID- 11020914 TI - Adverse effects of saline neurolysis on a chronically compressed rat sciatic nerve. AB - We have evaluated the effect of saline neurolysis compared with a simple decompression procedure on chronically compressed sciatic nerves in rats. Eight months after the initiation of nerve compression within a silicone tube, rats were divided into three groups of 35. In group I, saline was injected subepineurially at the compressed segment of the nerve after removal of the tube (decompression plus saline neurolysis). The nerves in group II were treated by removal of the tube only (simple decompression). In group III the tubes were left in place. Adding saline neurolysis to decompression provided no histological, morphometric, electrophysiological, or vascular advantages up to four months, and even gave worse results than simple decompression. We conclude that saline neurolysis has no beneficial effect on a chronically compressed nerve and it is not recommended for clinical use. PMID- 11020915 TI - Experimental model for local application of growth factors in skin re epithelialisation. AB - We did an experimental study to assess the effects of different growth factors on re-epithelialisation of skin wounds by creating a partial-thickness defect in rats with a handle dermatome. Three different growth factors that are particularly involved in the re-epithelialisation phase of wound repair (epidermal growth factor (EGF), keratinocyte growth factor (KGF), and basic fibroblast growth factor (FGF-b) n = 10 in each group) were applied locally in a hydrocolloid dressing containing solutions of the different factors (EGF 10 micrograms/ml, KGF 3.3 ng/ml, bFGF 1 microgram/ml). The dressings were changed daily. The thickness of the epithelium, the percentage of re-epithelialisation, and the maturity of the epithelium were quantified and measured morphometrically. The results showed that: the experimental model allowed us to apply the growth factors, while continuously maintaining the dose within the maximum activity of the growth factor; when EGF, KGF, and bFGF were given according to the protocol there was significant thickening of the new epidermis (p < 0.01) and acceleration of the re-epithelialisation (p < 0.05) rate compared with controls, and significantly more mature epithelium grew (p < 0.05) all of which were evident on both the third and the fifth days; and EGF and KGF cause a more epidermal thickening than bFGF. PMID- 11020916 TI - Microcirculation of a venous flap: an experimental study with microspheres in rabbits. AB - In recent years, it has been found that maintenance of venous circulation alone may support a small flap with no direct arterial inflow. The clinical application of a venous flap has potential in the field of microsurgery. The purpose of this study was to evaluate the haemodynamics within a pedicled venous flap in rabbits, compared with those of a composite graft. Pedicled venous flaps and composite grafts were raised from the abdominal walls of 30 adult New Zealand rabbits. Flap survival was measured and recorded and blood flow studies with microspheres were done for seven days. The viability of the pedicled venous flaps was much better than that of the composite grafts. At two weeks 24 of the venous flaps (80%) showed more than 75% surviving, but 29 (97%) of the composite grafts had less than 25% surviving. The results suggest that the blood flow through a patent vein maintained in a venous skin flap can provide enough nutrients for the flap to survive during the initial three days until neovascularisation. The venous flap receives more blood flow than a composite graft. We conclude that a venous flap depends on blood supply from the axial vein in addition to neovascularisation to maintain its survival. PMID- 11020917 TI - Maxillary dental arch and occlusion in repaired clefts of the secondary palate: influence of surgical closure with minimal denudation of bone. AB - A change in the method of surgical repair of cleft palate initiated this study of 64 patients with various degree of clefts of the secondary palate. Study casts were obtained at different ages. Certain measurements were taken and compared with those made in a previous investigation of similar patients operated on by the so-called "push-back" technique. Growth in the width of the maxillary dental arch and occlusion had improved, but there were significant differences only for the patients with velar clefts. PMID- 11020918 TI - A six-centre international study of the outcome of treatment in patients with clefts of the lip and palate: the results of a cross-linguistic investigation of cleft palate speech. AB - Speech samples of 131 subjects with complete unilateral clefts of the lip and palate from six European cleft palate centres were analysed and assessed using a specifically designed phonetic framework. This framework focused on consonants that are "vulnerable" in speech associated with cleft palate and common to the five languages of the project. The methodology used and the results of the reliability study are reported. Consonant articulation, resonance, and voice quality are also evaluated. The results show good outcomes with regard to consonant articulation across the whole study group with common areas of minor difficulty across languages. The results for resonance were less good, with slight hypernasality in 20% of subjects. There were, however, few indications of seriously disordered speech. The detectable differences between centres match the findings of the Eurocleft Orthodontic Group particularly in regard to the ranking of the centres. PMID- 11020919 TI - Active participation of mothers during speech therapy improved language development of children with cleft palate. AB - Whole language intervention uses the principles of natural language learning, which consider language not as an independent system but as a system intimately related to other cognitive and social abilities. This paper compares the outcome of speech therapy given in different settings to two groups of children with cleft palate. Those in the first group were treated by the speech pathologist alone (control group), whereas those in the second group were treated by the speech pathologist but were also accompanied by their mothers (experimental group). The purpose of this study was to find out if including the mother as an active participant in speech therapy sessions would improve the language development of children with cleft palate who also had additional language delays. Both groups were evaluated before and after treatment to evaluate the advance of each group. The patients accompanied by their mothers had significantly better language skills compared with patients treated without their mothers. The results support the statement that language development is related to mother-child mode of daily life interaction in children with cleft palate. PMID- 11020920 TI - Asymmetry of gait after free flap reconstruction of severe tibial fractures with extensive soft-tissue damage. AB - Gait patterns of the preferred speed were investigated on 17 patients recovering (9 months-14 years) after reconstruction of severe tibial fractures. A novel data acquisition system was used to record the plantar pressures as well as electromyographic (EMG) activities during walking. The results indicated incomplete recovery of symmetrical gait patterns. In particular, the duration of the stance phase was shorter on the operated side (mean (SD) 701 (90) ms compared with 765 (128) ms, p < 0.001). The peak pressure points under the foot were different on bilateral comparison, the operated side being regularly higher under the lateral forefoot area. This may imply attempts to reduce the loading of the ankle joint during stance. The pressure distribution models reflected these asymmetrical patterns more specifically than the EMG activities of the lower leg muscles examined. PMID- 11020921 TI - Trapeziometacarpal joint arthrodesis for the treatment of arthrosis. AB - We undertook a retrospective follow-up study of arthrodesis of 21 trapeziometacarpal joints in 19 patients. Internal fixation was by two or three Kirschner wires and a bone graft. No joint failed to unite. Painless joints and stable grasping were achieved. Subjective estimations of improvement were excellent for 16 joints. However, some patients complained of clumsiness after the procedure. Instability was seen in six metacarpophalangeal (MP) joints of the thumb. The most common indication for trapeziometacarpal joint arthrodesis was painful arthritis in vigorous, young male labourers. However, preoperative evaluation of the function of the MP joint is necessary to determine suitability for surgery. PMID- 11020922 TI - Follow-up of volar plate interposition arthroplasty (Tupper) of the metacarpophalangeal joints in rheumatoid hands: preliminary findings. AB - We present the results of a three-year follow-up study of 59 interposition volar plate arthroplasties (Tupper) on metacarpophalangeal joints in 13 patients with erosive rheumatoid arthritis. The median age at the time of operation was 60 years (range 45-77). All patients reported lasting pain relief at rest. Improvement of hand function was, however, less satisfactory as both grip and pinch strength were compromised in all patients. Seven patients were satisfied with the outcome whereas six complained of stiffness in the operated joints. All but one patient concluded that they would have the operation again if necessary. PMID- 11020923 TI - Weilby-Burton arthroplasty of the trapeziometacarpal joint of the thumb. AB - Twenty-three patients (25 thumbs) were treated by tendon interposition arthroplasty for trapeziometacarpal arthrosis as described by Weilby and modified slightly as described by Burton and Pellegrini. There was good (4/25, 16%) or complete (19/25, 76%) pain relief in 23 (92%) of the cases. Activities of daily living were generally easier. Mobility and strength of the thumb were satisfactory. One patient had signs of instability during a stress test. We conclude that our technique produces a stable and pain-free thumb joint. However, careful selection of the patients for this procedure is essential, and the patient must be given comprehensive information about all stages. PMID- 11020924 TI - Repeated digital nerve block for pain control after tenolysis. AB - We describe a way of achieving immediate painfree mobilisation after tenolysis or tenosynovectomy in Zone II. Bupivacaine is instilled along the flexor tendon sheath through a thin percutaneous catheter with an antibacterial filter. PMID- 11020925 TI - Complete breakage of three-dimensional miniplates: unusual complication of osteosynthesis after sagittal split osteotomy. Two case reports. AB - We describe two cases in which three-dimensional miniplates broke after bilateral sagittal split osteotomy. The miniplates broke vertically and the cause was suspected to be excessive shear force on the osteotomy line because of unstable occlusion. In patients with unstable postoperative occlusions the osteosynthesis should be bicortical. PMID- 11020926 TI - Superselective neurectomy with periorbital primary reconstruction for blepharospasm. Case report. AB - A 66-year old man with blepharospasm and ptosis of the brow was treated with a combined procedure in which the branches of the facial nerve were excised at the margin of the orbicularis, and the periorbital area was reconstructed simultaneously. The condition improved after the treatment with no occurrence of either oral complications or facial anaesthesia. PMID- 11020927 TI - Eradication of metastatic angiosarcoma of the lower extremity. Case report. AB - A 47-year-old man with an angiosarcoma of the right lower extremity was cured with mainly surgery and radiotherapy despite an inguinal metastasis. Histologically, the amputation stump of the metastatic tumour showed invasion by tumour. Radiotherapy, chemotherapy, and treatment with interleukin-2 was added. Ten years later he has had no recurrence. PMID- 11020928 TI - Use of the distally-based radial forearm flap supplied by the dorsal carpal arch, or palmar carpal arch, or both, in mutilating injuries. Two case report. AB - The distally-based radial forearm flap is safe, easy, and versatile as a regional, one-stage procedure to reconstruct soft tissue defects of the hand. However, there is a general perception that the deep and superficial palmar arches have to be intact to raise a distally-based radial forearm flap. We successfully used two flaps supplied by the dorsal carpal arch, or the palmar carpal arch, or both, despite the fact that the deep and the superficial palmar arches were damaged in mutilating injuries. PMID- 11020929 TI - True aneurysm of the digital artery. Case report and review. AB - We report a case of true aneurysm of the ulnar digital artery, the aetiology of which was related to gripping a golf club. It was diagnosed preoperatively by magnetic resonance imaging. The patient made a good recovery from excision, and the aneurysm had not recurred two years later. PMID- 11020930 TI - Isolated tuberculous tenosynovitis of the flexor tendon of the fourth finger of the hand. Case report. AB - A 21-year-old woman presented with a painful swelling of her palm which prevented her from fully extending her fourth finger. Magnetic resonance imaging showed synovial thickening around the flexor tendon and fluid in the tendon sheath. The mass was excised and histopathological examination showed tuberculosis. She had a six-month course of antituberculous treatment and was perfectly well three years later. PMID- 11020931 TI - The potential role of contraception in reducing abortion. AB - This study examines the potential role of further increases in contraceptive prevalence and effectiveness in reducing abortion rates. The model used in this analysis links the abortion rate to its direct determinants, including couples' reproductive preferences, the prevalence and effectiveness of contraceptive practice to implement these preferences, and the probability of undergoing an abortion to avoid an unintended birth when a contraceptive fails or is not used. An assessment of the tradeoff between contraception and abortion yields estimates of the decline in the total abortion rate that would result from an illustrative increase of 10 percentage points in prevalence. This effect varies among societies, primarily because the tendency to obtain an abortion after an unintended pregnancy varies. For example, in a population with an abortion probability of 0.5, a 10 percentage-point increase in prevalence would avert approximately 0.45 abortions per woman, assuming contraception is 95 percent effective. If all unintended pregnancies were aborted, this effect would be three times larger. Eliminating all unintended pregnancies and subsequent abortions would require a rise in contraceptive prevalence to the level at which all fecund women who do not wish to become pregnant practice contraception that is 100 percent effective. A procedure is provided for estimating this "perfect" level of contraceptive prevalence. PMID- 11020932 TI - Who's at risk? Men's STD experience and condom use in southwest Nigeria. AB - Many STD/HIV-prevention programs worldwide assume that individuals' risk of acquiring sexually transmitted diseases, including HIV infection, is highest in the context of commercial sex. To address this assumption, research conducted in urban Southwest Nigeria combined qualitative and quantitative methods to examine men's sexual behavior, condom use, and STD experience in different types of sexual relationships (marital, casual, and commercial). Logistic regression analysis of survey data indicates that number of sexual partners and sex with sex workers are positively and significantly related to STD experience. Follow-up in depth interviews with clients of sex workers indicate, however, that these men are actually more likely to report having contracted an STD from a casual sex partner than from a sex worker. Men are most uncertain about their vulnerability to STDs with casual partners. Men's condom use is highest in commercial sex, inconsistent in casual relationships, and lowest in marriage. STD/HIV-prevention programs need to address the range of sexual relationships and the meanings and behaviors associated with them. PMID- 11020933 TI - Trends and determinants of contraceptive use in Rakai District, Uganda, 1995-98. AB - A prospective study conducted between 1995 and 1998 assessed trends in contraceptive use in rural Rakai District, Uganda. Over a period of 30 months, women's use of modern contraceptives increased significantly from 11 percent to 20 percent. Male condom use increased from 10 percent to 17 percent. The prevalence of pregnancy among sexually active women 15-49 declined significantly from 15 percent to 13 percent. Women practicing family planning for pregnancy prevention were predominantly in the 20-39-year age group, married, better educated, and had higher parity than others, whereas women or men adopting condoms were predominantly young, unmarried, and better educated. Condom use was particularly high among individuals reporting multiple sexual partners or extramarital relationships. Contraceptive use was higher among women who desired fewer children, among those who wished to space or terminate childbearing, and among women with previous experience of unwanted births or abortions. Self perception of HIV risk increased condom use, but HIV testing and counseling had only modest effects. Contraception for pregnancy prevention and for HIV/STD prophylaxis are complementary. PMID- 11020934 TI - Sexual initiation and contraceptive use among adolescent women in northeast Brazil. AB - To estimate trends and determinants of sexual initiation and contraceptive use among adolescent women in Northeast Brazil, multivariate logistic hazard models are used that draw on data from three Demographic and Health Surveys conducted there between 1986 and 1996. Educational attainment is among the variables found to be associated most consistently with differential risk of engaging in first intercourse during adolescence, including premarital intercourse, and of contraceptive use during sexual initiation. Greater frequency of attending religious services and greater exposure to television are also associated with lower rates of sexual initiation and higher use of contraceptives. Seemingly diminishing returns of education on delayed sexual activity may help explain, in part, observed increases in the absolute level of adolescent sexual experience across survey periods, however. Multilevel modeling techniques pointing to the existence of cluster-level random variances underline the need for further research into community influences on individual sexual activity. PMID- 11020936 TI - Kenya 1998: results from the Demographic and Health Survey. PMID- 11020935 TI - The first generation to control family size: a microstudy of the causes of fertility decline in a rural area of Bangladesh. AB - In Bangladesh, the total fertility rate declined from more than six children per woman of reproductive age in 1980 to 3.3 children per woman in 1996. Much discussion has ensued about the circumstances responsible for this decline and, in particular, about the contribution made by the national family planning program. For this study, qualitative interviews were conducted with 67 women concerning the factors influencing their reproductive behavior. The participants, residents of a rural area in southwestern Bangladesh, consisted of two groups: women who were currently practicing family planning and women who had never practiced or had not done so for at least four years. The findings suggest that the family planning program has played an important role in reducing family size, but also indicate that economic and social changes, and especially growing aspirations (particularly for a life outside agriculture) have combined with changes in the nature of family decision making to make couples more receptive to the idea of family planning. PMID- 11020937 TI - Bolivia 1998: results from the Demographic and Health Survey. PMID- 11020939 TI - [Low pregnancy rate and small litters after artificial insemination]. PMID- 11020938 TI - [Veterinarians:'Watch your affairs!']. AB - The most relevant results of a written questionnaire among veterinarians in the Netherlands are presented and discussed. The inquiry was performed by MarketResponse Nederland BV. The objective was to get an overview of the current perception and the future view of the profession. The response was 37%. Most respondents (66%) were practitioners, 7% were active in research or teaching institutions, 5% were governmental employees, 5% were employed in industry, and 17% did not belong to any of these categories (retired, unemployed etc.). Forty seven per cent of the veterinarians practised mainly small animal medicine, 24% large animal medicine (cattle 15%, swine 8%, and poultry 1%), and 4% equine medicine; 24% worked in mixed practices. This division reflects the real-life situation. The percentage of female respondents was considerably higher in the group of recent graduates than in the other groups of graduates (increasing from 27% in the graduation period 1980-1989 to 56% in the period 1990-1999). Sixty per cent of the veterinarians worked more than 40 hours a week. Veterinarians considered themselves reliable, honest, professional, client-minded, and animal friendly. According to them, the public perceived veterinarians as being animal friendly, professional, and reliable. Veterinarians were less satisfied in their current position than other professional groups, particularly with regard to their income. Their current market position was considered indifferent or good. This position was influenced negatively by the decrease in the number of animals and competition from non-veterinarians and others. The situation could be improved by collaboration and practice fusions, specialization, and differentiation. More demanding and price-conscious clients and governmental regulations were considered important trends. Social, management, and marketing skills, increased knowledge and cooperation, and a vision of future developments were considered essential in order to be able react to developments on the market. About 60% of the practitioners and 50% of the other veterinarians felt that they lacked the right skills, and especially management and marketing skills, to react to these developments adequately. Most veterinarians (77%) considered that their training did not provide them with the skills needed for their current position. During training, more time should be spent on management, communication, marketing, and social skills. Based on the results, veterinary medicine in the Netherlands seems to be an introspective but good profession; however, the interesting aspects of the profession appear to be over-shadowed by less appreciated aspects and the income is moderate. Entrepreneuship is poorly developed, in part because this is considered taboo. The importance of skills that are not an integral part of veterinary medicine, such as management, marketing, and communication, is recognized, as is the lack of these skills. Despite this, emphasis is put on continuing professional education, species specialization, modernization, accreditation, and expansion of facilities and treatment possibilities as ways to respond to the market situation. PMID- 11020940 TI - [Predecessors: veterinarians from earlier times (40). Karl-Wilhelm Vix (1802 1866)]. PMID- 11020941 TI - [Grand opening of House Lameris]. PMID- 11020942 TI - [Development of veterinary medicine for the horse in the 20th century]. PMID- 11020943 TI - [Regulating industry control of poultry diseases]. PMID- 11020944 TI - The extent and impact of sheep pox and goat pox in the state of Maharashtra, India. AB - A survey of sheep and goat producers in the state of Maharashtra, India, was undertaken to ascertain the extent and economic impact of sheep pox and goat pox (SGP). One thousand one hundred and sixteen owners were interviewed. Eighty owners (7.2%) reported that they had experienced an outbreak of the disease in the previous 6 years. The results showed that, while producers ranked SGP below other infectious diseases such as foot-and-mouth disease, rinderpest and enterotoxaemia, when SGP occurred it had a major impact, with average morbidity and mortality rates of 63.5% and 49.5%, respectively. Modelling studies suggested it would take about 6 years for a flock or herd to recover from an outbreak, with average annual losses in income of 30-43%, depending on flock type and the owner's actions. Statewide, it is estimated that around 5000 flocks and herds are affected by SGP annually in Maharashtra, costing up to INR 107.5 million. The highest losses occurred in the Aurangabad region. PMID- 11020945 TI - Diversity of ectoparasites in sheep flocks in Sao Paulo, Brazil. AB - The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sao Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the 'Associacao Paulista de Criadores de Ovinos' (ASPACO; Sao Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host. PMID- 11020946 TI - Reproductive responses of Holstein-Friesian cattle to the climatic conditions of central Sudan. AB - Data were evaluated from a purebred Holstein herd (n = 1274) kept near Khartoum, a hot and dry area, from 1990 to 1996. The mixed model procedure in SAS was used to analyse the reproductive traits. The mean gestation length, days dry, number of services per conception, days from calving to first oestrus, days from calving to conception, calving interval and breeding efficiency were 279 days, 104 days, 4.2, 85.8 days, 208.9 days, 486.2 days and 74.9%, respectively. Factors that had significant effects (p < 0.01) on all the reproductive measures under study were the year and month of calving, and parity. The heritability estimates for all traits under investigation were zero. The repeatability estimates for the numbers of services per conception, days from calving to conception, days from calving to first oestrus and calving interval were 0.21 +/- 0.021, 0.12 +/- 0.024, 0.03 +/- 0.018 and 0.00, respectively. The phenotypic correlation for calving interval with numbers of service per conception was 0.05, that with days from calving to first oestrus was 0.06 and that with days from calving to conception was 0.07. Other correlations ranged from 0.00 to 0.28. The simple correlation coefficients between production and reproductive measures ranged from -0.06 to 0.45. PMID- 11020947 TI - Effects of supplementing a basal diet of Chloris gayana hay with one of three protein-rich legume hays of Cassia rotundifolia, Lablab purpureus and Macroptilium atropurpureum forage on some nutritional parameters in goats. AB - Growth and digestibility experiments were conducted on growing East African type goats offered Chloris gayana hay supplemented with one of three high-protein (119 128 g CP/kg DM) legume hays, Cassia rotundifolia (cassia), Lablab purpureus (lablab) or Macroptilium atropurpureum (siratro), and crushed maize to investigate the feed intake, digestibility, growth and urinary excretion of purine derivatives. Goats in the supplemented groups had higher total dry matter and nitrogen intakes and higher N retention and body mass gains than unsupplemented counterparts. The digestibility of dry matter, organic matter and neutral detergent fibre were increased by protein supplementation. Animals on supplemented diets had higher fractional outflow rates of particulate matter from the rumen. The production of protein by ruminal microbes and the efficiency of microbial N production were increased by supplementation. It was concluded that a mixture of low-quality grass hay (61.9 CP/kg DM) and either cassia, lablab or siratro hay, and maize grain can provide a productive balanced diet for growing goats. PMID- 11020948 TI - Growth and survival of Holstein and Brown Swiss calves reared outdoors in individual hutches. AB - The effects of outdoor raising of Brown Swiss and Holstein replacement calves in individual hutches were studied on a farm having northern Mediterranean climatic conditions. A total of 240 female calves was used with 30 calves of each breed and born in each of the four seasons. The animals were kept for 3 months in individual hutches and then in groups of 10 until 6 months of age. The birth weights of the Brown Swiss and Holstein calves were 37.2 +/- 0.21 and 37.9 +/- 0.22, respectively. The average birth weights were highest in spring and lowest in winter. Breed and seasonal effects were evident at 6 months. The live weights for the two breeds at 6 months were 114.1 +/- 1.3 and 126.3 +/- 1.5 kg, respectively. The numbers of health disorders were significantly higher in the first month of life. Mortalities occurred in the following months, being 2.5% in Holsteins and 5% in Brown Swiss by 6 months of age. The results indicated that calves can be reared outdoors successfully. PMID- 11020949 TI - Autoimmune systemic diseases. Proceedings of an international symposium. December 2-3, 1999. Ljubljana, Slovenia. PMID- 11020950 TI - Overview of the nomenclature and diagnostic categorization of vasculitis. PMID- 11020951 TI - Vasculitis: clinical approach, pathophysiology and treatment. AB - Systemic vasculitides constitute a heterogeneous group of diseases characterized by inflammation of blood vessels. Classification of the disease is mandatory for the assessment of prognosis and the institution of treatment. Secondary forms of vasculitis should be distinguished from primary vasculitides, as infections or other conditions underlying the secondary forms require a different approach. The presence of anti-neutrophil cytoplasmic antibodies (ANCA) directed against either proteinase 3 or myeloperoxidase strongly suggest one of the pauci-immune small vessel vasculitides. Their role in the pathogenesis of such vasculitides has been suggested by clinical and experimental data but has not been proven. Treatment of severe systemic vasculitides consists of cyclophosphamide in combination with corticosteroids but this regimen has many adverse effects. Less toxic regimens as well as new treatment modalities based on insight into the pathophysiology of systemic vasculitides have been developed and are currently being tested in multicentre European studies. PMID- 11020952 TI - Pathogenic role of anti-endothelial cell antibodies in systemic vasculitis. AB - Antiendothelial cell antibodies (AECA), a heterogeneous group of antibodies quite distinct from the ANCA family, have been detected in variety of diseases which share a varying degree of vessel wall damage. This review is mainly focused on Wegener's granulomatosis, Takayasu's arteritis and Kawasaki syndrome, which provide the best examples to evaluate the pathogenic and prognostic value of AECA. There is increasing evidence to show that AECA might be pathogenic in inducing autoimmune vascular disease. It is relevant to note that the presence and titre of AECA has been correlated with disease activity in systemic vasculitis. Experimental in vitro and in vivo models support a potential pathogenic role for AECA in sustaining immune-mediated vessel inflammation. Rather than being cytotoxic to endothelial cells, AECA are able to up-regulate the expression of adhesion molecules (E-selectin, intercellular adhesion molecule 1 and vascular cell adhesion molecule-1) and to induce the secretion of cytokine and chemokine which, in turn, cause leukocyte recruitment and adhesion. A recent idiotypic animal model has provided further evidence that AECA can be pathogenic. PMID- 11020953 TI - Antineutrophil cytoplasmic autoantibodies-antigen specificity and associated diseases. AB - Antineutrophil cytoplasmic antibodies (ANCA) are widely used as a useful diagnostic marker for small vessel vasculitides, although the test may occasionally be positive in various other conditions. The aim of this study was to assess ANCA in various clinical-pathological settings. ANCA were tested by indirect immunofluorescence and enzyme-linked immunosorbent assay and were found to be positive in 423 patients in the period from 1989-1999. Patients were grouped in accordance with their clinical-pathological setting as follows: 1. pauci-immune vasculitis confirmed by biopsy (n = 151), 2. clinically suspected vasculitis (n = 59), 3. inflammatory bowel diseases and autoimmune hepato-biliary disorders (n = 83), and 4. miscellaneous diseases (n = 130). The association of proteinase 3 ANCA with Wegener's granulomatosis (45/56) and myeloperoxidase ANCA with microscopic polyangiltis (45/54) and pauci-immune necrotising glomerulonephritis (24/28) was established. However, ANCA with other specificities were also shown to be present in these forms of vasculitides. ANCA, specific mostly for myeloperoxidase but also for other or unknown ANCA antigens, frequently revealing atypical immunofluorescence patterns, were characteristically found in other diseases. The titres of ANCA were significantly higher (p < 0.05) in patients with pauci-immune vasculitis than in those with clinically suspected vasculitis and other diseases. In conclusion, well standardised techniques for ANCA testing in conjunction with the clinical picture and histopathologic findings, if available, may significantly contribute to the diagnosis of small vessel vasculitides. PMID- 11020954 TI - ANCA-associated vasculitis--an autopsy study. AB - The purpose of the study was to analyse the autopsies of 31 patients (20 female, mean age 67 years, range 28-87 years; and 11 male, mean age 66 years, range 47-80 years) with antineutrophil cytoplasmic antibodies (ANCA)-positive vasculitis, which was clinically confirmed in 25 patients and suspected in 6 patients, who had been treated and had died between 1989 and 1999. Kidney biopsy was performed in 22 patients on average 33 months (range, 1-132 months) prior to death. Biopsy and autopsy tissue specimens were examined by standard light and immunofluorescence microscopy techniques. Pauci-immune extracapillary glomerulonephritis was found in nearly the same percentage of 22 renal biopsies and 31 autopsies, namely in 91% and 84%, respectively. Active necrotising extracapillary glomerulonephritis was the prevailing lesion in 75% of biopsies, while advanced sclerosing glomerular lesions prevailed in 69% of autopsies. In the biopsies, necrotising lesions predominated in patients with ANCA of proteinase 3 specificity, while sclerotic lesions were more often associated with myeloperoxidase-ANCA. In the autopsies, florid necrotising systemic vasculitis coexisted in 2 patients with advanced sclerosing glomerulonephritis. Autopsies revealed the actual expansion of vasculitic disease, disclosed clinically silent vasculitic involvement of unusual locations and, in 3 patients, confirmed the clinically suspected vasculitis. The final diagnoses in 31 patients were as follows: Wegener's granulomatosis (5 men, 2 women), microscopic polyangitis (10 women, 2 men), pauci-immune crescentic glomerulonephritis (4 women, 3 men), a single case each of polyarteritis nodosa and isolated cutaneous vasculitis. In 3 patients, suspected vasculitis was not confirmed at autopsy. Nineteen of 31 patients died from septic infections or necrotising pneumonias, and 6 patients from progressive or recurrent vasculitis with complications, altogether nearly 80%. Cardiovascular failure, including pulmonary thrombembolism, caused death in 6 patients. PMID- 11020955 TI - Wegener's granulomatosis in the upper respiratory tract. AB - Wegener's granulomatosis is a distinct clinico-pathological entity characterised by necrotising vasculitis of small arteries and veins in conjunction with the formation of granuloma in the upper and lower respiratory tracts, and glomerulonephritis. The vast majority of patients have antineutrophil cytoplasmic antibodies in the serum with a characteristic cytoplasmic pattern. However, in early phases of the disease only the upper respiratory tract may be affected, clinical and histological features may be nonspecific, and antineutrophil cytoplasmic antibodies not present. In this paper we present four patients with involvement of the upper respiratory tract suspicious for early Wegener's granulomatosis. We emphasise the significance of clinical, histological and serological parameters in the early detection of Wegener's granulomatosis. PMID- 11020956 TI - Wegener's granulomatosis--a diagnostic challenge. AB - Wegener's granulomatosis is a systemic vasculitis characterized by necrotizing granulomatous lesions in the upper and lower respiratory tracts, glomerulonephritis and vasculitis involving other organs. Limited forms have been described in which some features of the disease may be absent. Four patients with this disease are being reported with special emphasis on differences in presentation, the ensuing diagnostic problems, and individual outcome. In three, the disease began as a limited form with upper respiratory tract and eye involvement, while in one patient, onset was systemic including affection of the lower respiratory tract. The mean delay from first symptoms to diagnosis was 20 months--much longer for the three limited forms than for the one with systemic onset, in whom the condition was recognized after 2 months following initial misdiagnosis of respiratory and urinary tract infections. One patient developed endocarditis and required aortic valve replacement. Immunofluorescence revealed antineutrophil cytoplasmic antibodies in all, three showing a cytoplasmic pattern and antibodies to proteinase 3, and the fourth a perinuclear pattern and antibodies to myeloperoxidase. Upper respiratory tract biopsies were not specific. Kidney biopsies were performed in all the patients and were crucial for definitive diagnosis and treatment of the disease, which was successful in three patients. PMID- 11020957 TI - Autoantibodies and clinical subsets: relevance to scleroderma. PMID- 11020958 TI - Vascular complications in lupus nephritis and lupus-like syndromes. PMID- 11020959 TI - Correlation among WHO classes, histomorphologic patterns of glomerulonephritis and glomerular immune deposits in SLE. AB - In addition to the conventional World Health Organization (WHO) classification of lupus glomerulonephritis (GN), various concomitant approaches have been introduced in the evaluation of renal biopsies of patients with systemic lupus erythematosus (SLE) in order to increase the impact of biopsies on the decision concerning the most appropriate therapy as well as for establishing the prognosis. Three hundred and seventy kidney tissue samples from 267 SLE patients were analysed using standardised light, electron and immunofluorescence microscopic techniques. In 155 patients, a comparative clinical follow-up study and statistical analysis were performed. The study highlighted the heterogeneity of WHO classes IV and III, which include 5 and 6 different conventional histomorphologic types of GN, respectively. Mixed membranous and proliferative GN associated with "full-house" mesangial-transmembranous immune deposits, demonstrated in more than one third of our SLE cases, appears to be diagnostically most characteristic. Immune deposits distributed in the glomeruli in five different patterns, obviously play a major role in the pathogenesis of various WHO classes and histomorphologic types of lupus GN. Additional mechanisms related to the occurrence of antiphospholipid antibodies and antineutrophil cytoplasmic antibodies are suggested to contribute to the histomorphologic heterogeneity of WHO class III and IV lupus GN, particularly to the development of thrombotic, necrotising and crescentic glomerular lesions. In the present study, a statistically significant association was demonstrated between increasing mean values of the activity index and glomerular deposit distribution patterns labeled by subendothelial deposits. Furthermore, a significant correlation was established between an increasing risk of developing renal failure and increasing mean values of the chronicity index. Differences in the increasing risk of developing renal failure between groups with different histomorphologic types of GN and different immune deposit distribution patterns were not statistically significant. The surprisingly high renal survival rate of more than 80% noted in lupus patients with predominantly necrotising crescentic GN during the mean follow-up period of 40 months appears to be related to the more aggressive treatment of those patients. Our study confirmed a significant role of the WHO classification of lupus GN in the decision concerning the most appropriate treatment and prognostication. An increasing risk of irreversible renal failure in patients with WHO class IV lesions in contrast to those of WHO class III and in contrast to those of the category incorporating all other WHO classes was shown to be statistically significant. PMID- 11020960 TI - Tubulo-interstitial involvement in lupus nephritis with emphasis on pathogenesis. AB - Glomerular lesions in lupus nephritis have been extensively studied in recent decades, but much less attention has been paid to the tubulo-interstitial compartment. The aim of this study was to contribute to the understanding of the pathogenesis of tubulo-interstitial lesions in lupus nephritis by analysing their incidence, character, and their associations. One hundred and ninety kidney biopsies of 190 patients fulfilling American Rheumatology Association (ARA) criteria of systemic lupus erythematosus (SLE) were examined by traditional light, immunofluorescence and electron microscopy. Interstitial inflammatory infiltration and tubulo-interstitial immune deposits concurred in 72 cases (37.9%). Their frequency was the highest in WHO class IV lupus glomerulonephritis. By multivariate analysis, the intensity of interstitial inflammatory infiltration correlated best with the percentage of renal corpuscules with extracapillary crescents and the extent of interstitial fibrosis. On immunohistochemical assessment, the inflammatory infiltrate was found to be composed of CD45RO positive T lymphocytes (191.3/mm2), CD68 positive macrophages (101.7/mm2) and CD45RA positive B lymphocytes (17.2/mm2). For all cell types the median value was higher in cases with extracapillary crescents, and did not correlate with presence and intensity of tubulo-interstitial immune deposits. Infiltration showed the tendency of periglomerular distribution, especially around glomeruli showing extracapillary proliferation and destruction of the capsular basal membrane. Rare S100 positive cells were only found in the interstitium. Tubulo-interstitial lesions estimated semiquantitatively correlated with the degree of proteinuria. Our findings suggest that tubulo-interstitial deposits do not play a major role in the pathogenesis of tubulo-interstitial lesions. The formation of interstitial cell infiltrates appears to be greatly influenced by the development of extracapillary crescents, perhaps by direct transmission of the severe inflammatory process to the adjacent interstitium. The composition of the infiltrate, including antigen presenting cells may signalize an additional involvement of cell-mediated immune mechanisms acting against so far hypothetical tubular epithelial neoantigens. PMID- 11020961 TI - Small vessel thrombosis without major thrombotic events in systemic lupus erythematosus patients with antiphospholipid syndrome. AB - Antiphospholipid syndrome has been defined by the presence of antiphospholipid antibodies or lupus anticoagulant in association with certain clinical events, including recurrent arterial or venous thromboses and recurrent fetal loss. It comprises two separate clinical entities: simple, characterized by large vessel occlusions, and catastrophic, with multiple occlusive events predominantly affecting small vessels. Three patients with systemic lupus erythematosus and permanently increased IgG anticardiolipin antibody levels are being described. Only postmortem histopathological examination revealed microangiopathic thrombotic changes in different organs, which were clinically silent in early stages of the disease and misinterpreted later in its course because of a peculiar clinical picture. All patients presented features of catastrophic antiphospholipid syndrome in the final stage of the disease. PMID- 11020962 TI - Fingerprint and other organised deposits in lupus nephritis. AB - A prominent feature of lupus glomerulonephritis is extracellular, predominantly homogeneous electron dense deposits. Fingerprint-like deposits have been reported in 6 to 10% of cases. On electron microscopy, we studied the frequency and characteristics of organised deposits in 227 kidney tissue samples obtained by biopsy in 185 patients with systemic lupus erythematosus (SLE). Fingerprint forms of deposits were demonstrated in 34 biopsies of 32 patients (17.3%). In the control group of 626 kidney biopsies of patients with primary renal and systemic diseases other than SLE, no fingerprint deposits were found. In 227 kidney biopsy samples, fingerprint deposits were found to be associated with mesangial (8.8%), mesangial-subendothelial (3.8%), subepithelial (28.6%), mesangial-subepithelial (11.1%) and mesangial-transmembranous (19.4%) glomerular deposit distribution patterns. They were demonstrated more often at different locations along the peripheral capillary glomerular basal membrane (77.4%) than within the mesangial matrix (43.7%). In extraglomerular locations, fingerprint deposits were present in the interstitium in 8.8%, along the tubular in 23.5% and peritubular capillary basal membrane in 20.5%, in the wall of the arterioles in 64.7% and in the juxtaglomerular apparatus in 18.2% of biopsies. Organised fingerprint deposits consisted of semicircular dark and light lines, each with a diameter of about 10 to 15 nm. Unilaterally, spiky processes at a periodicity of 10 to 15 nm were seen. Among 14 of 185 SLE patients with cryoglobulinemia, fingerprint deposits were demonstrated in only 2 patients. We conclude that fingerprint deposits are characteristic, diagnostically relevant for SLE and represent morphologically a homogeneous group of organised deposits unrelated to cryoglobulins. In 3 SLE patients, 20 to 100 nm tubules and in 2 SLE patients, 10 and 18 nm Congo red negative fibrils were found. By their morphology and their structural characteristics, the tubules and fibrils resemble the tubules in primary immunotactoid glomerulopathy and fibrils in primary fibrillary glomerulonephritis. PMID- 11020963 TI - Role of lipids in the progression of renal disease in systemic lupus erythematosus patients. AB - Systemic lupus erythematosus (SLE) is an autoimmune connective tissue disease marked by immune-complex mediated lesions in small blood vessels of various organs, especially the kidneys, although other factors may also be implicated in the pathogenesis of the disease. This article focuses on the role of lipids in the progression of glomerular, vascular and tubulo-interstitial lesions in two patients with lupus nephritis associated with pronounced hyper- and dyslipidemia. The pathogenesis of progressive glomerulosclerosis in both patients appears to be multifactorial. In addition to immune complex mediated lupus glomerulonephritis, progressively active in the first patient, severe nephrotic-range persistent proteinuria, arterial hypertension associated with hyperfiltration and hyperperfusion injuries and, to a minor extent, hyper- and dyslipidemia were observed. Immunological and non-immunological factors were shown to contribute to the development of tubulo-interstitial lesions. In both patients, in addition to local immune deposits, prominent tubulo-interstitial lipid deposits were probably causally related to both hyperlipidemia and the increased permeability of the glomerular filtration barrier. Tubular lesions were highlighted by intracytoplasmic lipid droplets as well as small cleft-like spaces found to be impacted in the tubular lumina. They were seen to penetrate tubular epithelial cells and eventually lodge in the interstitium, surrounded by mononuclear cell infiltrates and foam cells. In both patients, hypertensive angiopathy and extraglomerular vascular immune deposits were demonstrated. In addition, in the second patient, arteriolar and small arterial hyaline was found at the age of 28 years to be full of lipids and calcium precipitates, suggesting a peripheral atherosclerosis-like process which never occurs as a natural age-related condition. In conclusion, all parts of the nephron may be involved in the pathogenetic process causally related or influenced by hyper- or dyslipidemia. Associated either with endothelial cell injury and consequent insudation of lipids in the vascular walls, glomerular filtration barrier injury with hyperfiltration, or tubulo-interstitial lipid deposition, the mechanism of tissue damage by lipids in all parts of the nephron shares similarities with the pathogenesis of systemic atherosclerosis. PMID- 11020964 TI - Immunoserological aspects of idiopathic inflammatory muscle disease. AB - In the present study, the autoantibody profile of 31 Slovenian patients with idiopathic inflammatory muscle disease was estimated: 11 with polymyositis, 11 with dermatomyositis--both groups diagnosed according to the criteria of Bohan and Peter--and 9 with myositis-overlap syndromes. Autoantibodies against most relevant muscle specific (Jo-1, Mi-2) and non-specific antigens (PM-Scl, U1RNP, native Ro, Ro60, Ro52, and La) were detected with one or more detection techniques: counter-immunoelectrophoresis, enzyme-linked immunoassay, immunoblot and immunoprecipitation, each using different antigen preparations (native, recombinant). With counter-immunoelectrophoresis using a native antigen substrate (rabbit thymus extract), we were able to detect anti-PM-Scl antibodies more readily than with other techniques, probably due to conformational epitopes of native PM-Scl. Patients with this serological profile constituted a distinct group, sharing features of polymyositis and systemic sclerosis. Compared to previously reported data, the greater frequency of anti-Jo-1 found in all groups of patients (64-87% for PM, 18-20% for dermatomyositis and 33-44% for overlap syndromes) was probably due to the various methods used and the different clinical characteristics of patients. The greater prevalence of anti-Mi-2 antibodies in dermatomyositis patients (67%) and in particular in polymyositis patients (33%) and myositis-overlap syndromes (33%) seemed to be mainly due to methodological differences. A strikingly high prevalence of anti-Ro52 positive patients with polymyositis (55%), dermatomyositis (22%), and myositis-overlap syndromes (33%) was demonstrated, but was detected by only one technique. Moreover, concurrence with anti-Jo-1 antibodies was noted (69%). PMID- 11020965 TI - Acute respiratory distress syndrome in a polymyositis patient with the anti-Jo-1 antibody. AB - We report a case of a 66-year-old caucasian woman suffering from polymyositis with the anti-Jo-1 antibody. Shortly after admission to our hospital, despite the fact that the patient was given intravenous pulse methylprednisolone (1000 mg/day for 3 consecutive days), she developed severe, ventilatory-dependent, acute respiratory distress syndrome. Evaluation for infectious or noninfectious aetiologies of acute respiratory distress syndrome was unrevealing. Bronchoalveolar lavage disclosed neutrophilic alveolitis. Histological examination of a transbronchial biopsy revealed an interstitial fibrosing process and cuboidalisation of the alveolar epithelium. In addition to high-dose methylprednisolone the patient was given intravenous pulse cyclophosphamide fortnightly for six weeks and afterwards every 4 weeks. All symptoms resolved and arterial blood gases returned to normal. Remission has been maintained with azathioprine. One year after onset, polymyositis is in complete remission. This is the first report of a patient suffering from polymyositis with the anti-Jo-1 antibody who survived such a complication. Intravenous pulse cyclophosphamide was probably a life saving therapy. PMID- 11020966 TI - Amyloidosis of the larynx. AB - Amyloidosis of the larynx is an uncommon disease and mainly a local occurrence. Hoarseness is the prevalent symptom. Surgical excision of the amyloid masses is the treatment of choice. In the present paper, the authors describe seven cases treated in the last twenty years. PMID- 11020967 TI - Recurrent sepsis and seronegative arthritis in a patient with a selective IgG3 deficiency. AB - In a sixty-one-year-old patient with chronic polyarthritis, two life-threatening septic events were observed over a period of 6 months. The patient also had a selective IgG3 deficiency. The susceptibility to infection and chronic polyarthritis observed in this patient were very likely a consequence of the selective IgG3 deficiency. PMID- 11020968 TI - [Validity of various obesity indices calculated from height and weight in adult females using the underwater-weighing method as a reference]. AB - We evaluated association between excess percent body fat (%Fat) and various obesity indices calculated from height and weight in 322 adult females. %Fat was measured by the underwater-weighing method, and obesity indices were based on the following 5 methods; Broca-Katsura method (Katsura method), Kato-Wataya method (Kato method), Japan Society for the Study of Obesity method (BMI method; based on the body weight at which BMI is 22), Meiji Life Insurance Co. method, and Table and Figure for the Assessment of Obesity and Leanness by the Ministry of Health and Welfare (MHW method). %Fat was 30% or more (obese) in 73 females (22.7%), 25-30% in 97 (30.1%), 20-25% in 88 (27.3%), and less than 20% in 64 (19.9%). The correlation coefficient between the obesity indices and %Fat were 0.71 for the Katsura method, 0.70 for the Kato-method, 0.72 for the BMI method, 0.70 for the Meiji Life Insurance Co. Method, and 0.63 for the MHW method, being significant for all methods (P < 0.01). When the cut-off point was set as 110% for each obesity index, sensitivity was the highest for the Katsura method (67.1%), and specificity was the highest for the Meiji Life Insurance Co. method (95.2%). Receiver operating characteristic (ROC) curves were similar in figure for the Katsura method, Kato-method, BMI method, and Meiji Life Insurance Co. method. For the MHW method, however, the curve was slightly farther from the point of sensitivity of 100% and 1-specificity of 0% than the others. Excess fat accumulation could not be accurately assessed by the obesity indices calculated from body height and weight. Validity was similar among the obesity indices except for the MHW method. PMID- 11020969 TI - [Case-control study of risk factors for cedar pollinosis among parents of 3-year old children]. AB - PURPOSE: To identify risk factors associated with cedar pollinosis among parents of three-year-old children. METHODS: The subjects were parents whose children underwent health examination at the age of three years in September and October 1997 in Tochigi prefecture. We distributed questionnaires to the examinees beforehand with the request that they be filled in and brought to the examination site. Information on parents' environmental factors (place of residence, type and structure of housing, and distance from roads with heavy traffic), and mothers' past history and family history of allergic diseases was obtained. We defined cedar pollinosis in terms of three symptoms, sneezing, nasal discharge, and nasal obstruction, between February and May. Controls were those who did not have any of the three symptoms. Environmental factors were assessed with univariate and multivariate analyses using unconditional logistic models. Maternal genetic factors were also analyzed with odds ratios and 95% confidence intervals calculated for each. We also compared odds ratios of environmental factors between groups with and without genetic factors. RESULTS: Of the parents of children taking the health examination, 90.2% took part (2,968 of 3,291 couples). Information was obtained from 2,846 mothers and 2,905 fathers. Mother and father cases were 312 and 229, and controls were 1,857 and 1,934, respectively. For the univariate analysis of environmental factors, place of residence (residential area/agricultural area), type of housing (apartment complex/solitary house), structure (reinforced concrete building/wooden house), and distance from heavy traffic (< 100 m/100 m+) were positively related to cedar pollinosis. For the multivariate analysis using unconditional logistic models, the odds ratio was significantly high for distance from heavy traffic (< 100 m/100 m+) among fathers. Mothers' past history and family history of allergic diseases showed high odds ratios for cedar pollinosis. We classified groups with and without past and family histories of allergic diseases (group with and without genetic factors) for assessment with multivariate analyses. Odds ratios for the group with an allergic history were higher than for those without such a history, but difference for factors such as, distance from heavy traffic type of housing, and structure were not statistically significant. CONCLUSION: Risk of cedar pollinosis increases with distance from heavy traffic among fathers. Mothers with histories of allergy show slightly elevated odds ratios for environmental factors, but without statistic significance. PMID- 11020970 TI - [Construction of a subjective fatigue scale for adolescent students]. AB - OBJECTIVE: There is no appropriate scale to evaluate subjective symptoms of fatigue (SSF) in everyday life in adolescent students. An existing scale long before constructed for laborers has been used. It is essential to establish a practical, rational scale. The purpose of this study was to construct a practical scale to measure subjective symptoms of fatigue for high school and college students. METHODS: A questionnaire on SSF was administered to 5,435 students aged 15-20 yr. Factor analysis based on the oblique Promax rotation method was applied to a correlation matrix consisting of 44 items. RESULTS: Six factors were interpreted as follows: difficulty with concentrated thinking, languor, reduced activation, reduced motivation, drowsiness and feeling of physical disintegration. A subjective fatigue scale (SFS-Y: 24 items) was constructed using these factors. The eigen-value of principal component analysis, alpha reliability coefficients, and test-retest reliability coefficients for each subscale were very high. CONCLUSION: The SFS-Y generated in the present study is a very practical scale with high validity and reliability. PMID- 11020972 TI - [Change in maternal bone mineral density during pregnancy and relationship between the density and foetus growth--a prospective study]. AB - In order to study change in maternal bone mineral density during pregnancy and its relationships with bone metabolism and maternal perinatal factors including foetus growth, bone mineral density and bone metabolic markers were measured in 45 pregnant women aged 26-35 years. Specifically, bone mineral density was measured twice, first at 8-20 weeks of gestation and secondly within two weeks postpartum by ultrasonic bone densitometry, while bone metabolic markers were assessed at 8-20 and 26-30 weeks of gestation and at one week postpartum. Bone mineral density and metabolic markers were also examined in 35 non-pregnant controls, twice with a six-month interval. The results of the measurements and subsequent examinations based thereon were as follows: 1. Stiffness as an index of bone density decreased significantly during pregnancy (mean: -4.3%), with wide variation among individuals (-20%-(+)11%). Stiffness in controls did not show any significant change within the 6 months. 2. During pregnancy, bone alkaline phosphatase and osteocalcin levels were significantly lower while urinary hydroxyproline levels (H.P/Cre) were significantly higher than in controls. Significant negative correlations were found between change rate in stiffness and HP/Cre measured at second trimester and postpartum. These results indicate that bone formation is reduced while bone resorption is increased during pregnancy, and that stiffness index reflects the extent of bone resorption. 3. Pre-pregnancy BMI and weight gain during pregnancy could not be linked with change in bone mineral density. 4. The women with greater bone density demonstrated a greater extent of loss postpartum. 5. Postpartum bone mineral density showed a significant, positive correlation with baby birth weight and height. Higher preservation of maternal bone mass is important not only for the mother's health but also for the baby's. Ultrasonic bone densitometry was found to be useful for measuring bone mineral density because it could detect small changes during pregnancy reflecting bone metabolism. PMID- 11020971 TI - [Follow-up study related to raising low-birth-weight infants relationship among physical growth, mental development and child-rearing anxiety]. AB - OBJECTIVE: In order to clarify relationship among physical growth, mental development of low-birth-weight (LBW) infants and child-rearing anxiety of mothers, a follow-up study was accomplished for LBW cases from birth to three years of age in a regional group. METHODS: A self-administered questionnaire survey was conducted for mothers whose premature infants were born from April 1992 through March 1993 and whose existence was confirmed from April 1995 through March 1996 in Fukui prefecture. A control group was selected from mature infants whose gestation was 37 weeks or more, who were born in the same year and in the same region. The questionnaire was focused on fetal and child growth, as well as child-rearing anxiety. RESULTS: Of the subjects, while including only 6% very LBW infants, the percentage of full-term LBW infants was 55.3% and that of pre-term LBW infants was 44.7%. In the former group, the mean birth weight (MBW) was 2,319 g, the percentage of females was 63.3%, firstborn children was 59.2%, and light for-dates (LFD) infants due to intrauterine growth retardation 76.9%. In the latter group, the MBW was 1,983 g, the percentage of females was 52.9%, that of firstborn children was 44.5%, and LFD infants was 14.3%. The growth of the LBW infants through the age of three fell into the same category of growth as the upper portion (1,250-1,499 g) of the very LBW infants rather than that of the mature infants. While the growth of appropriate-for-dates (AFD) infants at full term were normal, that of LFD infants and pre-term infants was insufficient, and the percentage of those demonstrating catch-up of mature infants was less than 80%. The group of pre-term infants was retarded in the gross motor skills, but not language understanding and pronunciation of two-word sentences. The child rearing anxiety in mothers who delivered pre-term infants was extremely severe after child birth. As the child grew older, however, the feeling of uneasiness was reduced. Mothers who were delivered of full-term LBW infants did not exhibit any fluctuation in child-rearing anxiety during the first three years. CONCLUSION: Fetal size affects growth progress of children. The prognosis of LBW infants is not satisfactory and mothers who deliver full-term LBW infants continue to worry about their child's growth. PMID- 11020973 TI - [Stroma supported erythropoiesis]. PMID- 11020974 TI - [Molecular diagnosis of hereditary sideroblastic anemia and model mouse of the disease]. PMID- 11020975 TI - [Erythroid stimulating factors]. PMID- 11020976 TI - [Erythropoietin receptor mutations]. PMID- 11020977 TI - [Transferrin receptor and erythropoiesis]. PMID- 11020978 TI - [Red cell membrane anomalies in childhood intractable anemia]. PMID- 11020979 TI - [Antithymocyte globulin treatment in 11 patients with aplastic anemia]. AB - We report the response to, and toxicity of antithymocyte globulin (ATG) treatment in 11 consecutive patients (7 men and 4 women; median age 46 years) with aplastic anemia (AA). Six of the patients had severe disease and 5 had moderate disease; all were treated within one year from diagnosis. The ATG regimen was the initial treatment for 6 patients, but a sequential treatment for the other 5. Cyclosporin A was administered orally 1-3 weeks after the ATG treatment. All patients were assessed for over 6 months (median, 20.4 months); 8 showed a good response, 1 a minimal response, and 2 no response. Disease severity had no influence on the response. In 1 patient, ATG treatment had to be discontinued because of hepatic toxicity. However, adverse reactions were not severe in the other 10 patients. These findings suggest that ATG treatment is a safe and effective therapy for AA. PMID- 11020980 TI - [Treatment outcome in elderly patients with aggressive non-Hodgkin's lymphoma]. AB - Seventy-one patients aged 61-84 years with previously untreated aggressive non Hodgkin's lymphoma were treated with a doxorubicin-containing regimen and evaluated retrospectively. The patients comprised 49 men and 22 women with a median age of 68 years. The median observation period was 544 days. Histological examination revealed 17 cases of diffuse small cleaved, 11 cases of diffuse mixed, 40 cases of diffuse large, and 3 cases of immunoblastic lymphoma, classified according to the International Working Formulation. When the patients were divided according to the age-adjusted international index, group A (61-64 years; n = 21) comprised 5 low (L)-, 4 low-intermediate (LI)-, 7 high intermediate (HI)-, and 5 high (H)-risk patients. The corresponding numbers in group B (> or = 65 years; n = 50) were 14, 12, 16, and 8, respectively. The overall three-year survival rate was 50%, being 78% in group A and 36% in group B (P = 0.02), and 77% for L + LI patients and 34% for HI + H patients (P = 0.003). The respective three-year survival rates for L + LI and HI + H patients were 100% and 67% in group A, and 68% and 16% in group B. HI + H patients in group B showed shorter survival than L + LI patients in group B (P = 0.002) and HI + H patients in group A (P = 0.03). The cause of death in most group B HI + H patients was lymphoma, although the dose intensity of doxorubicin, cyclophosphamide and vincristine did not differ significantly from that in the other groups. Thus, HI + H patients aged 65 and over had an essentially poor prognosis. PMID- 11020981 TI - [Prognostic significance of chromosome analysis in childhood acute lymphoblastic leukemia. Children's Cancer and Leukemia Study Group (CCLSG)]. AB - We analyzed the prognostic significance of chromosomal findings in children with acute lymphoblastic leukemia (ALL), treated according to the Children's Cancer and Leukemia Study Group protocols between 1987 and 1993. Patients were classified into 5 groups according to chromosome number. The patients with a hyperdiploid(> 50) karyotype(13%) had the best prognosis [4-year event-free survival (EFS): 83 +/- 6%], while those with a pseudodiploid karyotype (24%) had the worst prognosis(4-year EFS: 52 +/- 6%) (log-rank, p = 0.03). However, multivariate analysis revealed that the ploidy classification had no prognostic significance in terms of EFS. When patients were classified according to chromosome abnormalities, those with any type of translocation had a worse outcome (4-year EFS: 33 +/- 9%) than those with hyperdiploidy(> 50), normal diploidy, and other abnormalities(log-rank, p < 0.0001). Multivariate analysis revealed that chromosome abnormalities were an independent prognostic factor (relative risk 3.98; p < 0.0001). Patients with t(1; 19) had an EFS similar to that of patients with chromosome abnormalities other than translocations or normal diploidy. We conclude that chromosomal findings have prognostic significance, although some chromosome abnormalities lost their statistical significance after modern intensified chemotherapy. Childhood ALL should be further stratified according to chromosome classification. PMID- 11020982 TI - [Tacrolimus administration to a patient with cyclosporine-induced encephalopathy after allogeneic bone marrow transplantation]. AB - A 25-year-old woman with severe aplastic anemia received allogeneic bone marrow transplantation from an HLA-identical sibling. Pretransplant conditioning comprised 3.6 Gy of total body irradiation and 200 mg/kg cyclophosphamide. Cyclosporine (CSP) and methotrexate were administered to prevent graft-versus host disease (GVHD). The patient complained of severe headache soon after CSP administration on day-1. On day 3, convulsion developed and she lost consciousness for 15 min. CT and MRI demonstrated low density areas and high signals, respectively, in the frontal and parietooccipital lobes and splenium of the corpus callosum, suggesting brain edema probably induced by CSP. After immediate withdrawal of CSP, glycerol and prednisolone were instituted, and the patient's condition improved. Thereafter, she developed grade II acute GVHD. This was treated with tacrolimus, which produced no adverse effects including central nervous system (CNS) toxicity. This case illustrates that careful management of CNS disorders induced by CSP can be important in patients undergoing allogeneic bone marrow transplantation. PMID- 11020983 TI - [CD8-positive diffuse large B-cell lymphoma]. AB - In May, 1998, a 63-year-old woman was admitted for treatment of relapsed malignant lymphoma. In March 1997, a diagnosis of diffuse large B-cell lymphoma(DLBL), clinical stage IIISE A, was made from a biopsy specimen of the tumor in the left buccal mucosa. Six cycles of CHOP regimen were given, and complete remission was achieved. However, relapse of the lymphoma was suspected, and the patient was transferred to our hospital. On admission, a right tosillar mass, 8 mm in diameter, was found, and a biopsy showed DLBL. The immunophenotype of the lymphoma cells was CD3- CD4- CD5- CD8+ CD10- CD19+ CD20+ CD23- CD25+ IgL (lambda)+, and dual staining confirmed that the cells were CD8+ and CD19+. The patient was an HTLV-1 carrier, but monoclonal integration of HTLV-1 proviral DNA into the lymphoma cells was not detected. She was diagnosed as having CD8+ DLBL, clinical stage IA. Because she responded to salvage chemotherapy, autologous peripheral blood stem cell transplantation was performed and complete remission was obtained. To our knowledge, this is the first report of CD8+ DLBL. PMID- 11020984 TI - [Leukocytopenia and thrombocytopenia preceded by human parvovirus B19 infection: report of three adult cases]. AB - In Saiki City, Oita, Japan, erythema infectiosum in children has been prevalent from June, 1999 to the time of writing (January, 2000). We present three adult cases of parvovirus B19-associated leukocytopenia and thrombocytopenia that developed during this epidemic. Between June and November, 1999, a 32-year-old woman, a 38-year-old woman, and a 63-year-old man were referred to our hospital for treatment of leukocytopenia and thrombocytopenia. All complained of common cold-like symptoms. Their WBC counts (percentage of neutrophils) were 1,000/microliter (70%), 1,900/microliter (40%) and 1,680/microliter (40%), their hemoglobin levels 9.4 g/dl, 9.8 g/dl and 14.9 g/dl, and their platelet counts 10.8 x 10(4)/microliter, 6.9 x 10(4)/microliter and 4.5 x 10(4)/microliter, respectively. The diagnosis of parvovirus B19 infection was documented by the presence of B19-specific IgM antibodies or serum positivity for viral DNA. In two cases, the leukocytopenia and thrombocytopenia resolved gradually. In the other case, leukocytopenia, thrombocytopenia and B19 infection persisted for more than two months. These cases suggest that parvovirus B19 may be a common cause of leukocytopenia and thrombocytopenia even in adult patients without hematological disorders (erythropoietic stress), and that testing for parvovirus infection is justified in such patients, even if anemia is slight, especially when erythema infectiosum is prevalent. PMID- 11020985 TI - [MALT lymphoma of the larynx]. AB - We describe a 25-year-old Japanese woman with a MALT-type lymphoma of the larynx. She presented with a one-year history of hoarseness and increasing pain in the larynx. A small tumor was found on the left side of the false cord, and was biopsied under laryngoscopy in the department of laryngology. Histological examination showed the presence of centrocyte-like cells infiltrating the submucosa and forming lymphoepithelial lesions. The neoplastic cells were CD20+, CD79a+, and CD5-. Staining for keratin with CAM 5.2 highlighted the infiltrated epithelium. Analysis of DNA extracted from the biopsy specimen showed a clonal immunoglobulin heavy chain gene rearrangement, confirming the histological diagnosis of extranodal marginal zone B-cell lymphoma of the MALT type. To our knowledge, only 6 cases of MALT lymphoma of the larynx have been reported previously. The presence of MALT lymphomas arising at rare sites emphasizes the importance of accurate diagnosis and appropriate clinical management. Patients require careful periodic evaluation in order to time the therapy appropriately, and to avoid overtreatment and complications of therapy, including secondary malignancies. PMID- 11020986 TI - [Intravascular lymphomatosis diagnosed by bone marrow biopsy]. AB - Intravascular lymphomatosis (IVL) is a rare malignancy characterized by neoplastic proliferation of lymphoid cells within the lumina of small vessels. We report a case of IVL in a 69-year-old woman, who presented with pancytopenia and elevation of the serum LDH level. There was no skin eruption or neurological abnormalities. Clusters of abnormal lymphoid cells were barely evident in a peripheral blood smear. Laboratory examinations revealed high levels of LDH (2,602 IU/l) and sIL-2R (5,640 U/ml). Bone marrow aspiration revealed a normal cellular marrow with mild hemophagocytosis, but no tumor cells were detected. After admission, respiratory failure due to multiple pulmonary embolisms progressed, and continuous infusion of heparin had no apparent effect. Bone marrow vessels filled with lymphoma cells were observed in a biopsy specimen, thus establishing a diagnosis of IVL. Chemotherapy with the CHOP regimen was immediately instituted. The respiratory failure was dramatically improved, resulting in disappearance of the abnormal lymphoid cells from the bone marrow. After eight courses of CHOP, low-dose etoposide therapy was administered, and no symptoms of relapse were noticed. The diagnosis of IVL is difficult because it does not form masses of tumor cells. Bone marrow biopsy may be helpful for early diagnosis of IVL if the disease is suspected and searched for. PMID- 11020987 TI - [ALL complicated by obstructive jaundice due to choledocholithiasis after unrelated bone marrow transplantation]. AB - A 22-year-old man with acute lymphocytic leukemia underwent allogeneic bone marrow transplantation (BMT) from an unrelated donor in October 1996. In April 1997, he suddenly developed severe abdominal pain with nausea and vomiting. The diagnosis was obstructive jaundice associated with gallstones in the gallbladder and common bile duct. The patient underwent laparoscopic cholecystectomy and endoscopic removal of the stones in the common bile duct. The major component of the gallstones was bilirubinate calcium. Although the pathogenesis of gallstones after BMT remains unclear, several factors including impaired contractility of the gallbladder, hemolysis, changes in bile composition, and biliary tract infection may play important roles. PMID- 11020988 TI - [Secondary solid tumors in autologous-peripheral blood stem cell transplantation recipients]. AB - We investigated the incidence of post-transplant solid tumors in a cohort of 109 patients who had undergone auto-peripheral blood stem cell transplantation (PBSCT) for acute myelogenous leukemia (n = 18), acute lymphoblastic leukemia (n = 7), non-Hodgkin's lymphoma (n = 52), Hodgkin's disease (n = 5), multiple myeloma (n = 16), chronic myelogenous leukemia (n = 1), myelodysplastic syndrome (n = 3) and solid tumors (n = 7). The patients were followed up for a median of 32 months after PBSCT. Five second solid malignancies developed in 4 patients within 14 to 43 months after PBSCT: large cell carcinoma of the lung, adenocarcinoma of the rectum, cholangiocellular carcinoma of the liver, squamous cell carcinoma of the mouth, and adenocarcinoma of the gallbladder. The affected patients were 3 of 52 with non-Hodgkin's lymphoma and one of 16 with multiple myeloma. One of the patients was in the 5th decade of life (n = 31) and 3 were in the 6th decade (n = 31). The cumulative actuarial risk for developing post transplant solid tumors was 8% at 8 years. Elderly patients (> or = 60 year old) who have undergone PBSCT need to be observed carefully for second neoplasms because of their increased risk of post-transplant solid tumors. PMID- 11020989 TI - [First documentation of transfusion-associated babesiosis in Japan]. AB - A 40-year-old man received blood transfusion in December 1998 because of gastric bleeding from a peptic ulcer. One month later, he developed febrile hemolytic anemia. Administration of high doses of glucocorticoid significantly reduced the hemolysis, but did not cure the disease. To investigate the cause of the hemolysis, the patient was transferred to our hospital in May 1999. Giemsa stained peripheral blood smears showed Babesia parasites in the red blood cells (RBC), and PCR analysis confirmed the presence of Babesia microti DNA. The parasitemia disappeared hematologically after 2 weeks of quinine and clindamycin therapy. However, parasite DNA was still detectable in the RBC. Although treatment with oral atovaquone was given for 2 weeks, parasitemia and febrile hemolysis recurred within a month after the last treatment. Fortunately, complete remission was obtained after a second 12-week course of therapy with quinine and clindamycin. PCR analysis revealed asymptomatic Babesia infection in one of eight samples from the original blood donor. The initial steroid therapy given to the patient without an accurate diagnosis seemed to have delayed augmentation of the specific antibodies (IgG) against Babesia microti, thus prolonging the parasitemia after the initial acute stage of babesiosis. PMID- 11020990 TI - [Aggressive transformation and extramedullary tumor formation in IgA-lambda multiple myeloma]. AB - A 52-year-old woman complained of lower back pain and gluteal pain in April 1997, and was found to have anemia, hypercalcemia and renal disorder. In September of the same year, she was diagnosed as having IgA-lambda myeloma (stage IIIA). VMMD IFN therapy was started in November, 1997, and this resulted in improvement of the M-protein level, and relief of the pain in the lower back and gluteal region. A second course of VMMD-IFN therapy was also effective. In April 1998, however, the back pain worsened, and in July the patient suffered a fall and fractured her left femur. Upon readmission to our hospital, the level of M-protein was lower, and high fever, hypercalcemia, renal disorder, elevation of the LDH level, anemia and thrombocytopenia were observed. Bone marrow examination revealed 30% atypical large-sized CD19-, CD38+, CD56+ myeloma cells and chromosomal abnormalities. Although the symptoms were improved temporarily after a third course of VMMD therapy, disease aggravation occurred again, and extramedullary masses appeared on the head, face and pelvis. VAD therapy was performed without effect, and the patient died about 2 months after recurrence. This was a comparatively rare case of fulminant multiple myeloma occurring in the terminal stage. PMID- 11020991 TI - [Non-Hodgkin's lymphoma in two married couples]. AB - Couple 1: A 74-year-old woman was diagnosed as having diffuse large B-cell lymphoma (DLBL) by left axillary lymph node biopsy. About 6 months later, DLBL was also diagnosed in her 79-year-old husband by right submandibular lymph node biopsy. Although the wife achieved partial remission with chemotherapy, she died due to disease progression. The husband's disease was chemotherapy-resistant, and he died of renal failure. Couple 2: An 86-year-old man was diagnosed as having DLBL by left axillary lymph node biopsy. About 4 years later, DLBL was also diagnosed in his 86-year-old wife by left axillary lymph node biopsy. Both the husband and the wife received chemotherapy. The husband is currently alive in complete remission, and although the wife achieved partial remission, she died due to disease progression. In both of these couples, it was considered unlikely that Epstein-Barr virus or human T-cell lymphotropic virus type I was related to the development of non-Hodgkin's lymphoma, and no environmental factors were confirmed to be involved. It is postulated that other unknown factors or agents may be associated with the development of lymphoma in married couples. PMID- 11020992 TI - [Pure red cell aplasia induced by clomipramine hydrochloride in a patient with SLE]. AB - A 48-year-old woman, who had been suffering from systemic lupus erythematosus (SLE), developed normochromic normocytic anemia after receiving clomipramine hydrochloride. Her reticulocyte count was low, and a bone marrow aspirate revealed erythroid hypoplasia without involvement of other cell lines. Thus a diagnosis of pure red cell aplasia (PRCA) was made. The anemia gradually resolved following withdrawal of the drug. Although several drugs are known to cause PRCA, this is the first time that clomipramine hydrochloride has been reported to have such an effect. The underlying SLE in this case suggested the possible immunological pathogenesis of drug-induced PRCA. PMID- 11020993 TI - [Granulocytic sarcoma presenting as an epidural mass with spinal cord compression]. AB - A 73-year-old man was admitted because of back pain and paralysis of the lower extremities. Magnetic resonance imaging of the spine at the Th4-6 level, obtained after gadolinium injection, demonstrated abnormal signal intensity within the Th5 6 vertebral bodies and an extradural soft-tissue mass on the right posterior side of the spinal canal, compressing the thecal sac. The patient underwent prompt decompression with laminectomy, but this was unsuccessful. A biopsy sample of the mass revealed the histological features of granulocytic sarcoma, including diffuse infiltration of numerous cells containing cytoplasmic granules and immunohistochemical positivity for myeloperoxidase. Two months later, a subcutaneous soft-tissue mass appeared at the anterior chest wall, and this was confirmed to be granulocytic sarcoma by microscopic examination. Both of these tumors were radiosensitive, but the patient died of septic shock. Granulocytic sarcoma usually occurs in association with leukemia or other myeloproliferative disorders. However, it is rarely found before leukemia becomes evident in the peripheral blood or bone marrow; only eight such instances have been reported previously. PMID- 11020994 TI - [MALT lymphoma producing IgG-kappa type M-protein]. AB - A 72-year-old woman, who has been administered prednisolone and azathioprine with diagnoses of idiopathic thrombocytopenic purpura (ITP) and autoimmune hepatitis (AIH), underwent a complete medical examination because of monoclonal gammopathy (IgG-kappa). Tumors were found in the ileum and descending colon. Pathological examination of biopsy specimens suggested a diagnosis of marginal zone B-cell lymphoma of the MALT type with a high-grade component. Flow cytometric analysis by two-color staining revealed that the neoplastic B cells expressed CD38, CD19, IgG and kappa, but not CD5 or CD10. There were no abnormal plasma cells in bone marrow smears. The patient achieved complete remission after receiving three cycles of THP-COP chemotherapy, which resulted in a decrease of the IgG level to within the normal range. These findings indicated that monoclonal IgG-kappa might be produced by lymphoma cells. However, the relationship of the immunosuppressive agents to the pathogenesis of the MALT lymphoma remains to be clarified. PMID- 11020995 TI - [Progression of refractory thrombocytopenia to chronic myelomonocytic leukemia accompanied by various inflammatory reactions]. AB - A 51-year-old man was admitted for treatment of severe thrombocytopenia in May 1997. A diagnosis of MDS RA (refractory thrombocytopenia; RTC) was made by bone marrow examination, which revealed mild marrow hypoplasia and a reduced number of megakaryocytes accompanied by micromegakaryocytes and hypolobular megakaryocytes. Chromosome analysis demonstrated 46, XY, t(5;7) (q31;q22) in all 20 cells examined. The patient received only supportive therapy including platelet transfusion, until leukocytosis and monocytosis gradually developed in November 1998. In view of a marked increase in the number of monocytes (more than 3,000/microliter), a diagnosis of CMML was made in December 1998. As the leukocytosis progressed, various inflammatory symptoms such as facial erythema and endophthalmitis developed. Administration of interferon alpha (IFN alpha) unexpectedly worsened the leukocytosis and monocytosis, suggesting abnormal responses of these cells to IFN alpha. Detailed molecular analysis of these cells might reveal a novel mechanism of leukemogenesis associated with 5q31. PMID- 11020996 TI - [Autologous transplantation of Ph-negative peripheral blood stem cells for treatment of chronic myelogenous leukemia]. AB - A 21-year-old man, diagnosed in March 1997 as having chronic myelogenous leukemia (CML), received hydroxyurea followed by daily interferon (IFN) until December 1998, when the additional chromosome abnormality of +8 appeared. As no suitable matched donor was available, the patient received mobilization therapy consisting of mini-ICE (idarubicin, cytarabine, etoposide) followed by G-CSF subcutaneously. During hematopoietic recovery, a total of 12 x 10(6)/kg CD34-positive cells were harvested. Cytogenetic analysis of peripheral blood stem cell (PBSC) products using FISH revealed 1% BCR/ABL fusion signals. In March 1999, he received conditioning therapy consisting of busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg) followed by infusion of 5 x 10(6)/kg CD34-positive cells. A neutrophil count of 500/microliter and a platelet count of 5 x 10(4)/microliter were attained by days 20 and 38, respectively. Bone marrow aspirates showed 2.6% BCR/ABL fusion signals on day 35 after autologous PBSC transplantation, and the patient remained in chronic phase until the sixth month, when a cytogenetic relapse (Ph, +8:4/20) occurred. These observations suggest that Ph-negative progenitor cells can be harvested using a mini-ICE regimen followed by G-CSF, and that autologous PBSC transplantation is feasible in patients with CML resistant to IFN. PMID- 11020997 TI - [Successful treatment of refractory thrombotic thrombocytopenic purpura with cyclosporine A and splenectomy]. AB - A 19-year-old girl was admitted to our hospital because of general fatigue, headache and purpura. A number of her laboratory findings suggested hemolytic anemia and thrombocytopenia. Direct/indirect Coombs tests gave negative results. Although the patient had no neurological or renal abnormalities, peripheral blood smears showed marked red cell fragmentation, and therefore she was diagnosed as having thrombotic thrombocytopenic purpura (TTP). Fresh frozen plasma (FFP) was transfused daily. The thrombocytopenia and hemolysis immediately improved, but worsened again after reduction of the FFP transfusion. Plasma exchange was instituted every other day, but the patient's condition worsened. Palsy and consciousness disturbance developed, and finally she lapsed into a coma. Daily plasma exchange and methylprednisolone pulse therapy were performed, together with weekly vincristine therapy, and this led to a gradual improvement in the patient's condition. However, several attempts at weaning from plasma exchange resulted in exacerbation of the TTP activity. Therefore oral cyclosporine A was started and splenectomy was performed. After these interventions, despite transient relapse, the patient was successfully weaned off the FFP transfusion, and she is now in remission. Because in this case splenectomy and cyclosporine A resulted in sustained remission of TTP that was refractory to intensive plasma therapy, an autoimmune mechanism may have been involved in the pathogenesis. PMID- 11020998 TI - [Successful hematologic reconstitution using CD34+ cells positively selected from cryopreserved autologous peripheral blood stem cells in a patient with malignant lymphoma]. AB - Clinical use of CD34+ cells positively selected from cryopreserved peripheral blood stem cells (PBSC) has been limited, and there have been only a few reports of this procedure, mainly because clump formation decreases the proportion of CD34+ cells that can be recovered. A 49-year-old Japanese woman with non Hodgkin's lymphoma (NHL) (follicular mixed, B cell, stage IVA) was treated with seven cycles of conventional chemotherapy and achieved partial remission. During hematopoietic recovery after the seventh course of chemotherapy, PBSC were harvested by continuous leukapheresis and cryopreserved. However, clonal rearrangement of the immunoglobulin heavy chain gene was detected in the PBSC by Southern blot analysis. After high-dose chemotherapy, CD34+ cells were positively immunoselected from the cryopreserved PBSC and infused into the patient at 1.97 x 10(6)/kg. The overall purity and recovery rate of the CD34+ cells were 72.2% and 65.0%, respectively. There were no severe adverse effects after PBSC transplantation, and the time required for recovery of neutrophils to over 0. 5 x 10(9)/l and platelets to over 50 x 10(9)/l was 11 and 21 days, respectively. Transplantation of CD34+ cells positively selected from cryopreserved PBSC provides engraftment ability similar to that of unmanipulated PBSC. PMID- 11020999 TI - [Paralytic ileus during treatment of acute promyelocytic leukemia with all-trans retinoic acid]. AB - A 54-year-old man was referred to our hospital because of petechiae and pancytopenia. Bone marrow aspiration showed a normocellular marrow with 92.4% promyelocytes. PML/RAR alpha mRNA was detected by reverse transcription polymerase chain reaction. On the basis of above data, a diagnosis of acute promyelocytic leukemia (APL) was made, and treatment with all-trans retinoic acid (ATRA) at a dose of 60 mg/day was begun. Fourteen days after the start of treatment, the patient developed paralytic ileus, accompanied by hyperleukocytosis, high fever, renal dysfunction and elevation of the serum FDP level. There was no evidence of infection. At this time, retinoic acid syndrome was suspected, and therefore steroid pulse therapy was started, which led to an improvement of the symptoms within four days. This case suggests that ATRA may have an adverse effect on the small intestine, causing paralytic ileus. PMID- 11021000 TI - Treatment of childhood non-Hodgkin's lymphoma. PMID- 11021001 TI - Immune function of small bowel and its related diseases. AB - The small intestine is a major immunological organ with aggregated lymphoid tissue of Peyer's diffuse lymphoid tissue of lamina propria and intra-epithelial lymphocytes as 3 key components. Immunopathogenesis is of central importance for coeliac disease, Crohn's disease and gastrointestinal food allergy. Small intestinal enteropathy of variable severity, related to T-cell activation is common to all. PMID- 11021002 TI - Hypovolemia and hypovolemic shock in children with nephrotic syndrome. AB - Hypovolemic shock is not an uncommon presentation in nephrotic syndrome, yet it is seldom mentioned in the literature. This study was performed to investigate the prevalence of hypovolemia and hypovolemic shock in the acute nephrotic stage, and the association of hemoconcentration and abdominal pain with hypovolemic status. Two hundred and twenty-five patients with a total of 328 admissions to the pediatric ward of our hospital during 1983 to 1996 were retrospectively reviewed for hypovolemic episodes. Clinical presentation and laboratory data including hemoglobin, serum sodium, albumin, cholesterol, and triglyceride levels were investigated. Thirteen patients with 19 episodes (5.8%) of hypovolemic shock were found, and had more severe hemoconcentration (hemoglobin 19.6 +/- 2.2 g/dL) and hyponatremia (127.3 +/- 7.2 mEq/L). Another 33 patients with 41 symptomatic hypovolemic episodes without hypotension (12.5%) were found, and their hemoglobin levels were higher compared to patients without hypovolemic symptoms. Among 61 episodes of abdominal pain and hemoconcentration, 58 were responsive to albumin infusion. This suggested the presence of hypovolemia. Twenty patients had abdominal pain without hemoconcentration, and 18 of them had primary peritonitis. Hypovolemia was found in patients at the acute nephrotic stage, and was usually associated with hemoconcentration and abdominal pain. A combined examination of hemoglobin and serum sodium is the best indicator of hypovolemic status. Both primary peritonitis and hypovolemic episodes should be taken into consideration when managing abdominal pain in children with nephrotic syndrome. PMID- 11021003 TI - Effect of ice water ingestion on asthmatic children after exercise challenge. AB - Both exercise and ice water ingestion are known to be trigger factors for an asthma attack in ethnic Chinese asthmatic children. The purpose of this study was to investigate whether ice water ingestion further deteriorates pulmonary function of asthmatic children after exercise. Thirty Chinese asthmatic children underwent exercise challenge by ergocyclometer for 6 minutes and then were further challenged by immediate ingestion of ice water (200 ml, 0-4 degrees C), warm water (200 ml, 37 degrees C) or no ingestion on three different days in one week. Each patient completed the three different water ingestion tests after exercise challenge. The FEV1, FEF25-75%, and PEF tests were performed at baseline and again at 5, 15, 30, 60, 90 minutes after exercise plus water ingestion challenge. After the spirometric test at 90 minutes, 3 puffs (0.6 mg) of hexoprenaline from a metered dose inhaler were given and then a further spirometric test was performed 15 minutes later. The FEV1 and PEF were significantly decreased after exercise plus the 3 different water ingestion challenge except for the FEV1 in the patients who ingested nothing (p = 0.051) and PEF in the patients who ingested warm water (p = 0.163). FEF25-75% of the three tests was not significantly decreased. Exercise-induced asthma (EIA) developed in about two thirds of the 30 patients, regardless of whether ice water, warm water or nothing at all was ingested after exercise challenge. There was no statistically significant difference in spirometric data among the 3 different water tests at various time points. The mean percentage increase of FEV1, FEF25-75% and PEF after bronchodilator therapy were all the lowest in the ice water test, and the greatest in the warm water test. A statistically significant difference was found between ice water and warm water tests for FEV1 and PEF (p = 0.0293 and p = 0.0308 respectively). In conclusion, about two thirds of the asthmatic children in this series had EIA. Those who ingested warm water after exercise had a better bronchodilator response than those who ingested ice water. PMID- 11021004 TI - Clinical experience in the treatment of ulcer-induced gastric outlet obstruction in seven children. AB - Peptic ulcer disease complicated with gastric outlet obstruction is rare in children. The experience in the treatment in children is limited. From 1987 to 1998, 7 children with gastric outlet obstruction due to peptic ulcer disease were encountered in our hospital. The 5 patients treated from October 1987 to May 1997 underwent truncal vagotomy, combined with pyloroplasty in 4 patients and antrectomy in 1. The indications for operation were failure of treatment with a histamine 2 (H2)-receptor antagonist. The remaining 2 patients, treated from June 1997 to October 1998, were successfully treated with oral omeprazole with or without the addition of antibiotics. These 2 patients had shorter hospital stays and could tolerate feeding earlier than those treated with truncal vagotomy. Although surgical treatment has merits in the management of children with peptic ulcer disease complicated with gastric outlet obstruction, in light of our cases we disclosed the trend toward intensive medical treatment. PMID- 11021005 TI - Treatment results of the TPOG-NHL92 protocols for childhood non-Hodgkin's lymphomas in Taiwan: a report from the Taiwan Pediatric Oncology Group (TPOG) AB - A nation-wide chemotherapeutic trial for childhood non-Hodgkin's lymphoma (NHL) was conducted by the Taiwan Pediatric Oncology Group (TPOG). Four TPOG-NHL92 protocols based on stage and histology were activated in 1992: TPOG-92LD (treatment duration: 8 months) was used for localized (stages I/II) NHL with any histology, 92LB (2 years), 92SNC (5 months), and 92LC (1 year) for advanced (stages III/IV) lymphoblastic (LB), small non-cleaved cell (SNC), and large cell (LC) lymphoma, respectively. From January 1992 through June 1998, 200 children with newly diagnosed NHL from 13 member hospitals of TPOG were enrolled. There were 140 boys and 60 girls. Their ages at diagnosis ranged from 2.4 months to 18.3 years with a median of 8.2 years. There were 54 (27.3%) patients with LB, 94 (47.5%) with SNC including B-cell acute lymphoblastic leukemia (B-ALL), and 50 (25.2%) with LC. Stages I, II, III, and IV (including B-ALL) of the disease comprised 5%, 10%, 43%, and 42% of cases, respectively. There were 176 patients eligible for evaluation of treatment results. The remission rate of induction was 82.4%, induction failed in 22 (12.5%) patients, and nine patients died during induction. As of August 31, 1999, 26 patients relapsed, six died during remission, one patient developed secondary acute myelomonocytic leukemia, and 105 patients remained in continuous remission with a median remission duration of 49 months. The event-free survival (EFS) at 7 years was 63.5%, 61.5% and 65% for LB, SNC, and LC groups (P = 0.8298). The 7-year EFS for stages I/II, III, and IV of the disease was 73%, 68.9%, and 50.3% (P = 0.0212), respectively. We concluded that following the strategy of stratification of therapy, only disease stages had prognostic significance in this study. More efforts are needed to improve our treatment results. PMID- 11021006 TI - The follow-up on the training course of pediatric advanced life support. AB - Two parts are included in this article. First, to improve the training course in pediatric advanced life support (PALS), we held a 2-day PALS training course on March 28 and 29, 1999. A pretest, posttest, and survey questionnaire were given to each participant. Of the 143 participating in the 2-day PALS course, 137 completed the study. The mean pretest score was 70.2 (SD = 11.5) and the mean posttest score was 86.9 (SD = 9.1). There was significant improvement noted after the 2-day PALS training course (p < 0.001). The 2-day PALS training course resulted in greater improvement in scores compared with the previous 1-day PALS training course in some certain groups of participants. The number of participants complaining that they were too rushed to get the best learning result declined from 71 of 106 (66.3%) taking the 1-day training course to 43 of 137 (31.4%) taking the 2-day training course. Second, a same written test was given to the 129 participants who had finished a 1-day PALS training course one year previously to evaluate the participants' retention of knowledge after 1 year. Of 82 responders who took the 1-year follow up test, 75 were from the medical center. The score of the follow-up test for these 75 participants showed a significant decline when compared with the posttest, except for those receiving another training course of advanced life support during the follow-up year. We conclude that the 2-day PALS training course is better than the 1-day PALS training course. Those who participate in the pediatric critical or emergent care had better receive the 2-day PALS training course. The participants receiving the 1-day PALS training showed a significant decline in their knowledge after 1 year, and they should reinforce their knowledge in the following years. PMID- 11021007 TI - Bilateral ulnar hemimelia in Brachmann-de Lange syndrome: report of one case. AB - Twenty-three cases of Brachmann-de Lange syndrome (BDLS) have been reported in literature here in Taiwan, but none of them had severe upper limb anomalies. We report on a male infant with BDLS who has bilateral ulnar hemimelia. PMID- 11021008 TI - Subglottic hemangioma associated with cutaneous and cerebellar hemangiomas detected by MRI: report of one case. AB - Subglottic hemangioma (SGH) is a benign neoplasm that may cause severe and life threatening respiratory obstruction in infants. However, patients usually present with inspiratory stridor in the first few months of life and may be mistakenly diagnosed as recurrent or persistent croup. Definitive diagnosis is made by image studies, endoscopic examination and biopsy or all. We report a 2-month-old female infant of SGH with initial clinical manifestations of dyspnea and inspiratory stridor co-existing with cutaneous and cerebellar hemangiomas. Clinicians must be alert the possibility of SGH when associated with cutaneous hemangioma. This patient has received oral steroid treatment for more than two months with improvement of the airway obstruction. Although purplish patch lesions over left side of face, eyelid, cheek, and peri-oral regions regressed, the size of the SGH on the followed MRI was slightly enlarged. The diagnosis and various treatments of SGH are discussed and reviewed in this paper. PMID- 11021009 TI - Hereditary fructose intolerance presenting as Reye's-like syndrome: report of one case. AB - Hereditary fructose intolerance (HFI) is an autosomal recessive disease caused by catalytic deficiency of aldolase B (fructose-1, 6-bisphosphate aldolase). Herein we report on a case of hereditary fructose intolerance with initial presentation of episodic unconsciousness, seizure, hypoglycemia, hepatomegaly, and abnormal liver function since the patient was 11 months old. She was diagnosed as Reye's like syndrome according to a liver biopsy done at 20 months of age. As she grew up, cold sweating, abdominal pain or gastrointestinal discomfort shortly after the intake of fruits was noted and she developed an aversion to fruits, vegetables and sweet-tasting foods. At 9 years of age, a fructose tolerance test signified a positive result that induced hypoglycemia, transient hypophosphatemia, hyperuricaemia, elevation of serum magnesium, and accumulation of lactic acid. Appropriate dietary management and precautions were recommended. The patient has been symptom-free and exhibited normal growth and development when followed up to 12 years of age. PMID- 11021010 TI - Congenital diaphragmatic hernia misdiagnosed as pneumothorax in a newborn. AB - Congenital diaphragmatic hernia (CDH) is usually left sided and has a large defect allowing abdominal viscera herniated into thoracic cavity. The chest films usually show air-filled stomach and/or loops of bowel in the ipsilateral hemithorax with mediastinal shift. We report a newborn with CDH, presenting as hyperlucent hemithorax, right-shifted mediastinum, apparently normal pattern of abdominal bowel gas, with the tip of nasogastric tube below the left hemidiaphragm on the radiograph. It was initially misdiagnosed as pneumothorax, and the acute respiratory distress was temporarily relieved by needle aspiration. Hyperlucent hemithorax due to intrathoracic gastric dilatation alone is an unusual presentation of CDH in neonatal period. Absence of stomach bubble in the left upper quadrant of the abdomen, in both radiography and abdominal sonography, is an important clue to make diagnosis of CDH in this misleading condition. PMID- 11021011 TI - [Skin care and topical steroids]. PMID- 11021012 TI - [Current concepts of asthma in adolescence: from the standpoint of internal medicine]. PMID- 11021013 TI - [In view of allergy in 21st century from 20th--bronchial asthma]. PMID- 11021014 TI - [Clinical efficacy of immunotherapy with house dust in the patients with perennial nasal allergy]. AB - We tried to elucidate the relation of age and severity with the efficacy of immunotherapy. The subjects included 51 patients with perennial allergic rhinitis who had immunotherapy for more than a year. Symptom scores, serum total IgE levels, Dermatophagoides pteronyssinus specific IgE, and peripheral blood eosinophils counts were evaluated before, half a year, one year and two years after the onset of immunotherapy. 64% of the children's group and 69% of the adults' group of patients showed significant improvement in their nasal symptoms, showing that age is not a factor influencing the efficacy of immunotherapy. The severity of the disease influenced the clinical efficacy; the more severe the disease, the poorer the efficacy of the immunotherapy. Periphipheral blood eosinophils counts gradually decreased after the onset of immunotherapy. High eosinophil counts before immunotherapy predicted poor efficacy of the immunotherapy. PMID- 11021015 TI - [The role of lipid mediators in bronchial hyperresponsiveness and airway eosinophil accumulation induced by antigen challenge in guinea pigs]. AB - The aim of this study was elucidate the role of lipid mediators in bronchial hyperresponsiveness (BHR) and airway eosinophil accumulation 24 hours after an antigen challenge in guinea pigs. Thromboxane (TX) A2 receptor antagonist, S-1452 (1, 10 mg/kg), cysteinyl leukotriene (cLT) receptor antagonist, ICI-198, 615 (0.5, 5 mg/kg), platelet activating factor (PAF) receptor antagonist, E-6123 (1, 10 micrograms/kg), and each vehicle were intraperitoneally given 1 h before and 11 h after an ovalbumin (OVA) challenge. BHR to inhaled methacholine was measured and then bronchoalveolar lavage (BAL) was performed 24 h after the OVA challenge. The three drugs significantly inhibited BHR to methacholine, dose dependently. S 1452 significantly inhibited total cell counts (TCC). ICI-198, 615 significantly reduced both TCC and eosinophil percentage, but E-6123 did not alter TCC and cell differentiation in BAL fluid. Therefore, these results clearly showed that lipid mediators were involved in antigen-induced BHR and suggested that TXA2 and cLT may contribute to the penetration of inflammatory cells through capillary wall, still more cLT is concerned eosinophil accumulation with cell specificity. PAF dose not take part in the penetration of inflammatory cells. PMID- 11021016 TI - [Effect of a leukotriene antagonist, pranlukast hydrate, on exercise-induced asthma for 14 days]. AB - Exercise is a common stimulus of bronchoconstriction in subjects with asthma. Exercise-induced asthma (EIA) may be caused by heat loss and/or water loss during exercise. These changes cause the release of mediators and the development of bronchoconstriction. Recent studies have implicated the cysteinyl leukotrienes in the pathogenesis of EIA. Therefore we investigated the effect of a leukotriene antagonist, pranlukast hydrate, on EIA. Subjects were 11 EIA positive patients and were all atopic type bronchial asthma. 450 mg of pranlukast hydrate was given for 14 days and the exercise test was performed before and after treatment. Pulmonary function tests were done before and 0, 5, 15 and 30 min after completion of the each exercise. Prostaglandin E2, thromboxane B2, leukotriene E4 and B4 in urine were measured before and after the treatment. Pranlukast hydrate did not affect FEV1.0, PF and Flow volume curve at rest and also levels of mediators in urine. The mean maximal percentage fall in FEV1.0 after exercise was -54.9 +/- 5.7% without therapy and--30.1 +/- 7.5% after pranlukast hydrate therapy. There was a significant difference between FEV1.0 values before and treatment at each time (p < 0.05). This study demonstrated that pretreatment with a potent and selective leukotriene antagonist, pranlukast hydrate, attenuated EIA, and it suggests that leukotriene plays a role in EIA. PMID- 11021017 TI - [Atopic dermatitis at 18-months physical examination]. AB - In order to know the prevalence rate and severity of atopic dermatitis, and to compare them to those previously done for 4-month-old infants, the authors examined atopic dermatitis of 18-month-old children at regular health checks at a health center. Among 769 children examined, 69 were diagnosed as having atopic dermatitis, giving a prevalence rate of 9.0%. Most of cases were mild, and no severe cases were observed. In contrast to the location of eruptions on head, face, and trunk at 4-month-old, the eruptions were seen mainly on the trunk and extremities. The prevalence rate in summer and autumn was significantly lower than that in winter and spring. Among 501 children whom we could examine both at 4-month and 18-month-old, 102 cases were diagnosed as having atopic eruptions at 4 months. The eruptions, however, were disappeared in 83 cases when they were 18 month-old. The severity of atopic dermatitis also came to be mild when they grew up. Altogether, a quarter of children had atopic eruptions either at 4 months or 18 months, or both. It will be of interest to observe the transition of atopic dermatitis in childhood and we are planning to follow them at next physical examination at 3-year-old. PMID- 11021018 TI - [Effect of a patch formulation of tulobuterol on cough scores and PEF in patients with cough variant asthma]. PMID- 11021019 TI - Hidden agendas? PMID- 11021020 TI - Medical emergencies. PMID- 11021021 TI - Quick fix? PMID- 11021022 TI - Orthodontics on TV. PMID- 11021023 TI - Orthodontics on TV. PMID- 11021024 TI - Oral cancer screening. PMID- 11021025 TI - Oral cancer screening. PMID- 11021026 TI - NHS direct. PMID- 11021027 TI - HRT ulceration. PMID- 11021028 TI - Room with a view. PMID- 11021029 TI - Piercing dangers. PMID- 11021030 TI - Introduction to acupuncture in dentistry. AB - An introduction to the practical application of acupuncture in dentistry is presented in the light of current research. It is concluded that acupuncture could supplement conventional treatment modalities. Its value in the treatment of temporomandibular dysfunction syndrome and facial pain has been well documented and supported by randomised controlled trials. Although it may be useful in the control of post-operative pain, its use as sole analgesia for operative care is questionable. The mode of action of acupuncture can be explained with reference to modern neurophysiology. A short training course can allow the technique to be an effective tool in every dentist's hands. PMID- 11021031 TI - The treatment of adult patients with a mental disability. Part 2: Assessment of competence. AB - Over recent years practitioners are increasingly being asked to attend to, or to provide treatment for, adult patients with some degree of mental illness, either in their homes or in the dental surgery. Because the issue of competence and the assessment of the patient is crucial to this aspect of dental practice this second paper is devoted to an overview of that assessment process; outlining the legal tests and standards to be employed. PMID- 11021032 TI - Social background of minority ethnic applicants to medicine and dentistry. AB - AIM: To explore ethnic variations in social background of successful applicants to undergraduate United Kingdom medical and dental schools. METHOD: Retrospective analyses of University and College Admissions Services data on all students to commence study in pre-clinical medicine and dentistry, during the academic years 1994/5, 1995/6 and 1996/7. Analyses were undertaken for two categories of social class, namely higher (professional and intermediate) and lower (skilled non manual, skilled manual, partly skilled, and unskilled) social class. RESULTS: Over 15 thousand students were accepted to study medicine and dentistry during the three-year study period, of which 80% were from high social class backgrounds. More medical (80.9%) students were from high social class backgrounds than dental (73.3%) students (OR = 1.54, 95% CI = 1.39, 1.70). Social class differences were observed, with a greater proportion of higher social class students amongst the white students than amongst the minority ethnic students (OR = 1.42, 95% CI = 1.30, 1.55). This was more marked in dentistry (OR = 1.48, 95% CI = 1.22, 1.79) than in medicine (OR = 1.35, 95% CI = 1.22, 1.49). More students from higher social class backgrounds were observed in medicine than in dentistry amongst the black (OR = 1.55, 95% CI = 0.59, 4.00), Indian (OR = 2.04, 95% CI = 1.58, 2.62) and white (OR = 1.44, 95% CI = 1.26, 1.64) groups. CONCLUSIONS: Significant inter-ethnic differences are observed in the social background of students entering medicine and dentistry. Dentistry accepted a greater proportion of students from lower social class backgrounds and from black and minority ethnic groups. PMID- 11021033 TI - Dental care for children and young people who have a hearing impairment. AB - AIM: The aim was to determine whether there are indications that hearing-impaired children experience difficulties in accessing dental care and/or in receiving dental treatment. METHOD: The study was carried out by means of a questionnaire. Parents of 84 children contacted through the National Deaf Children's Society returned completed questionnaires. RESULTS: Eighty-two children (98%) had visited a dentist. Nearly two-thirds (63%) were reported to have at least one problem in communication while receiving dental care, this increased significantly as the severity of the hearing impairment increased. Fifty-nine children (70%) reported having at least one problem in communication at the doctors'. Fifty-two (62%) reported that the dentist had worn a mask while communicating with the child and 48 (57%) that there had been background noise in the surgery during appointments. CONCLUSIONS: Removing masks while talking, reducing background noise and learning to use simple signs may improve communication with hearing-impaired children. PMID- 11021034 TI - The use of interprofessional peer examiners in an objective structured clinical examination: can dental students act as examiners? AB - OBJECTIVE: To assess whether final year dental students could act as reliable examiners within an Objective Structured Clinical Examination (OSCE) by comparison with results obtained by an experienced member of staff. DESIGN: A station testing examination of the mouth was included in the second year medical undergraduate summative OSCE examination. SETTING: Concurrently run in three different examination venues on the Ninewells Hospital campus. SUBJECTS: 147 medical students and 3 pairs (A, B, C) of examiners. Each examining pairing consisted of one member of staff and one dental student (blind to each other's marking). METHOD: A checklist of 13 tasks to be performed was provided to the examiners. One mark awarded for a completed task, no mark for no attempt at the task, and half a mark for attempt at task. RESULTS: Paired results were available for 125 medical students. Using Mann-Witney analysis, the non-parametric 95% confidence intervals for the difference in scores between the 3 paired teams were group A (-0.5, 0), group B (-0.5, 0.5), group C (-0.5, 0). In only 4 students (out of 125) did the difference between the individual pair differ by 2 or more marks. CONCLUSION: On the basis of this pilot study final year dental students may be used as examiners in OSCEs where basic technical skills are to be evaluated. This development from peer group teaching provides further evidence supportive of interprofessional education. PMID- 11021035 TI - Concreteness and relational effects on recall of adjective-noun pairs. AB - Extending previous research on the problem, we studied the effects of concreteness and relatedness of adjective-noun pairs on free recall, cued recall, and memory integration. Two experiments varied the attributes in paired associates lists or sentences. Consistent with predictions from dual coding theory and prior results with noun-noun pairs, both experiments showed that the effects of concreteness were strong and independent of relatedness in free recall and cued recall. The generally positive effects of relatedness were absent in the case of free recall of sentences. The two attributes also had independent (additive) effects on integrative memory as measured by conditionalized free recall of pairs. Integration as measured by the increment from free to cued recall occurred consistently only when pairs were high in both concreteness and relatedness. Explanations focused on dual coding and relational-distinctiveness processing theories as well as task variables that affect integration measures. PMID- 11021036 TI - The effect of divided attention on memory for items and their context. AB - Memory for context, in comparison to memory for items, is a more demanding task and requires more attentional resources. We examined differences between item and context memory using divided attention at encoding and retrieval. Participants were presented with word lists and were instructed to learn the items (i.e., words), the intrinsic context (i.e., the colour of the cards on which each word was presented), and the extrinsic context (i.e., the temporal order of the words). Among 72 young adults, in comparison to conditions of full attention, divided attention applied at encoding only or retrieval only resulted in equally lower performance on all memory tasks; in contrast, divided attention applied at both encoding and retrieval resulted in lower performance only on memory for temporal order. The findings support the idea that memory for temporal order requires greater attentional resources and strategic processing than memory for items. PMID- 11021037 TI - The contribution of nonvisual information to simple place navigation and distance estimation: an examination of path integration. AB - In Experiment 1, participants walked without vision to a target location they had either previously viewed, were led to and from blindfolded, or both viewed and were led to and from blindfolded. A course to the target could be set and held without vision only if prior vision of its location was available. The locomotor group reproduced the heading and distance to the target less accurately than the other groups, which did not differ significantly. However, when nonvisual information accompanied vision of the target location, it served to subtly influence performance. Participants in Experiment 2 estimated the distance of a target they either viewed or were led to blindfolded. When vision was available, men overestimated target distance and women underestimated it. When target distance was learned nonvisually, no sex differences in distance estimations emerged. Our findings suggest that deriving navigation information from nonvisual locomotion is difficult and may be dependent on prior visual information. PMID- 11021038 TI - The role of the fixation location in inhibition of return. AB - Experiments that have examined inhibition of return (IOR) have typically used trial sequences in which attention is reoriented to a central fixation location between the presentation of the peripheral cue(s) and the target. In examining the role of the fixation location in IOR, three experiments were conducted in which an exogenous cue was used to reorient attention following a peripheral cue and before the appearance of a target. However, this cue occurred at either the traditional central fixation location or a nonfixated location. The results indicate that reorienting attention to a fixated location results in a significant reduction in the inhibitory effect. The results from the study suggest that IOR could serve as a mechanism that improves the efficiency of visual searches. PMID- 11021039 TI - Lexical access codes in visual word recognition: are the joint effects of context and stimulus quality diagnostic? AB - Is it possible to identify when lexical access in visual word recognition is based on an orthographic code and when it is based on a phonological code? Some researchers have argued that the joint effects of semantic context and stimulus quality in lexical decision are diagnostic. Their argument is that when phonological recoding is employed, it serves to keep the effects of stimulus quality and context from interacting. Semantic context and stimulus quality are therefore predicted to have additive effects on RT. In contrast, when lexical access is not mediated by a phonological recoding stage, then the effect of stimulus quality interacts with context, as commonly seen in the literature. A strong test of these claims was devised in which participants were forced to use phonological recoding for the purpose of lexical access. An interaction between context and stimulus quality was observed. This finding is taken as evidence that the joint effects of semantic context and stimulus quality are not diagnostic with respect to the nature of the lexical access code (i.e., orthographic versus phonological) for readers of English. PMID- 11021040 TI - [The development of experimental pharmacology 1790-1850]. AB - 1. The use of drugs goes back to the origins of mankind. In historical times oral drug-lore became codified empiric drug theory (materia medica) and ultimately, in the 19th century, experimental pharmacology. The initiator of experimental pharmacology as an independent medical discipline is Rudolf Buchheim (1820-1879). This study traces the pathways leading to Buchheim and identifies his predecessors between 1790 and 1850. The history of empirical pharmacology and its major theories in Antiquity, the Middle Ages, and early modern times is summarized. For the 18th century an overview is given on early attempts at experimental testing of drug effects and on the new therapeutic systems and medical sects. 2. Many authors have dealt with the grievances of pharmacology and therapy between 1790 and 1850, among them chief representatives of contemporary medicine like the French Fourcroy, Bichat, Pinel, Alibert, Magendie, and the Germans Schonlein, Mitscherlich, Wunderlich, Henle, and Oesterlen. Their criticisms are a means for a better understanding of the situation. They cover the following aspects. Pharmacology is distorted by speculations on the causes of drug action and confusion with regard to terminology and indications. Drug actions are being tested with inadequate methods. An increase in the number of drugs is mistaken for an increase in knowledge. The statement is made that pharmacology is the least developed of all medical subjects. The critics point out that only a more developed chemistry, physiology, and etiology will allow a scientific pharmacology. The drug theories of the medical sects are likewise rejected. Polypharmacy, composite drugs, and absurd formulas are regarded with contempt. Aggressive drug therapy is repudiated, but this easily results in avoidance of drugs and in therapeutic nihilism. 3. In 1799 Johann Christian Reil elaborated his principles for a future pharmacology. Reil establishes the rules for clinical experiments on which a scientific pharmacology should be based. His goal is to explain the actions of drugs which are the results of biochemical alterations. Even though Reil's program is a theoretical conception, it anticipates a situation that was to take shape half a century later. Also in 1799 Adolph Friedrich Nolde published detailed rules for the critical examination of drug actions in patients, including aspects like placebo, compliance, statistics, and several ethical rules. Reil's and Nolde's programmatic messages vanished in the emerging German medicine of "Naturphilosophie". 4. In the decades after 1800 medicine was at its zenith in the Paris School. It became a hospital medicine, based on anatomy and pathology. Francois Magendie was one of its representatives. He started out as a physician in 1808 and became a physiologist who soon surpassed his teachers Bichat and Richerand. Magendie's sole interest were facts, which had to be unravelled by experiments, mainly on animals. He created modern physiology based on the laws of physics and chemistry. Nevertheless, he remained an outsider among the Paris School. Bichat and other predecessors of Magendie had considered an experimental pharmacology based on physiology, however, they did not provide knowledge resulting from experiments. Magendie published his first experimental study of a pharmacological problem in 1809. From then on he studied the mechanism and site of action of drugs and used them at the same time as tools for the investigation of physiological processes. After Serturner's isolation of morphine from opium the preparation of pure alkaloids became a specialty of French pharmacists and chemists. Magendie sought their collaboration from 1817 on, convinced that pharmacology and therapy must be based on both physiology and chemistry. In 1821 he published his Formulaire pour la preparation et l'emploi de plusieurs nouveaux medicamens which marks the beginning of modern pharmacology. It grew throughout eight editions up to 1835. (ABST PMID- 11021041 TI - Privatization = poverty. PMID- 11021042 TI - Trading away public health: WTO obstacles to effective toxics controls. PMID- 11021044 TI - Bearing witness for tobacco. PMID- 11021043 TI - Tobacco control and direct democracy in Dade County, Florida: future implications for health advocates. AB - In 1979 and 1980 in Dade County, Florida, a small grassroots advocacy group, Group Against Smoking Pollution (GASP), attempted to enact a clean indoor air ordinance through the initiative process. The tobacco industry's successful efforts to defeat the initiatives were expensive high-tech media-centered campaigns. Even though GASP's electoral resources were extremely limited for both initiatives, GASP utilized similar media-centered tactics. This approach attempted to defeat the tobacco industry in its own venue, in spite of the tobacco industry's vastly greater resources. Nevertheless, the industry defeated these ordinances by narrow margins because of broad voter support for the initiatives before the industry started its campaigns. Health advocates will never have the resources to match the tobacco industry in expensive high-tech media-centered initiative campaigns. Rather, their power lies in the general popularity of tobacco control legislation and their ability to mobilize broad grassroots efforts combined with an adequately funded media campaign. PMID- 11021045 TI - Public opinion on alcohol policies in the United States: results from a national survey. AB - We surveyed the U.S. non-institutionalized population age 18+ on opinions regarding 23 alcohol control policies (N = 7,021). The cooperation rate among contacted households was 70% and the overall response rate was 54%. Results showed high levels of public support for most alcohol control policies. Over 80% support restrictions on alcohol use in public places, such as parks, beaches, concert venues, and on college campuses. Eighty-two percent support increased alcohol taxes, provided the funds are used for treatment or prevention programs. Over 60% support alcohol advertising and promotion restrictions, such as banning billboard advertising, banning promotion at sporting events, or banning liquor and beer advertising on television. Multivariate regression analyses indicated significant relationships between alcohol policy opinions and a variety of sociodemographic, political orientation, and behavioral measures. However, the absolute differences in alcohol policy support across groups is small. There is a strong base of support for alcohol control policies in the U.S., and such support is found among whites and ethnics of color, young and old, rich and poor, and conservatives, moderates, and liberals. PMID- 11021046 TI - Local option policies and alcohol availability in North Carolina counties. AB - Concerned about social problems related to alcohol, communities are considering legal alternatives for restricting its availability. Whether restrictive local policies are realistic means for curtailing the physical availability of alcohol is unknown. Using North Carolina's 100 counties as the unit of analysis, the extent to which 1994 local option policies predicted 1995 rates of alcohol permits, controlling for other factors related to the demand for alcohol, was examined. Counties with restrictive policies were found to have lower rates of alcohol permits than counties with permissive policies, after accounting for alcohol demand. Local option policies may be a reasonable strategy for limiting the physical availability of alcohol. In turn, limiting alcohol availability may be effective in reducing alcohol-related community problems such as traffic crashes and violent assaults. PMID- 11021047 TI - Oregon's Toxic Household Products Law. AB - In 1991, Oregon became the first state in the U.S. to require the addition of an aversive agent to ethylene glycol-containing antifreeze and methanol-containing windshield wiper fluid. This new law, entitled "Toxic Household Products (THP) Act," was designed to reduce pediatric and animal poisonings from accidental ingestion of these two potentially lethal consumer automotive products. While not the stated intention of the law, addition of aversive agents to consumer automotive products could also reduce adult poisonings associated with intentional (suicides or alcoholics ingesting methanol-containing windshield wiper fluid) or accidental exposures. This law went into effect April 30, 1995, following settlement of a lawsuit brought by the Chemical Manufacturing Specialties Association (CSMA), a trade group representing the five largest manufacturers of ethylene glycol-based antifreeze in the U.S. This paper discusses the major policy issues that arose following the passage of Oregon's THP Act. Major provisions of the law are provided along with a discussion of CSMA's opposition to the Act's implementation. A description of the eventual settlement that was reached with CSMA as well as the major components of Oregon Health Division's (OHD) enforcement program are also highlighted. Data are presented for 1987 through 1998 on the number of exposures and severity of effects for pediatric cases (children < 6 years old) following exposure to both of these potentially lethal automotive products. However, because of the low incidence of exposures each year, these data are insufficient to draw any conclusions on the impact of the THP Act. PMID- 11021048 TI - Manipulation method for the treatment of ankle equinus. AB - Ankle equinus is a well-known clinical entity that has previously been shown to compound a variety of foot and ankle conditions. Treatments for this disorder have included surgery to lengthen the Achilles tendon and daily stretching. This article describes a method of manual manipulation that can immediately and substantially increase ankle joint dorsiflexion. Patients treated with manipulation in the current study demonstrated nearly twice as much dorsiflexion motion as that demonstrated by patients in a prior study who were treated with a 5-minute daily stretching program for 6 months. PMID- 11021049 TI - The effect of rearfoot eversion on maximal hallux dorsiflexion. A preliminary study. AB - Functional hallux limitus has been recently described as an etiologic factor in postural complaints. Eversion of the rearfoot has been theoretically linked to decreased hallux dorsiflexion, although there have been no scientific studies on this subject to date. This study was undertaken to investigate the relationship between rearfoot eversion and hallux dorsiflexion; the results show that hallux dorsiflexion is decreased with rearfoot eversion in a static setting. PMID- 11021050 TI - Techniques for obtaining specimens for culture to confirm onychomycosis. AB - A study was conducted to determine whether fungal culture results in cases of suspected onychomycosis differ depending on the location from which the specimen is obtained. Specimens were taken from the nail plate, subungual debris, and nail bed in 30 patients with clinical evidence of onychomycosis. Cultures from the subungual debris were more likely to be positive for dermatophytes, nondermatophytic molds, and yeasts than were cultures from the deeper nail bed or nail plate. PMID- 11021051 TI - Nontraumatic foot complaints in older people. A population-based survey of risk factors, mobility, and well-being. AB - In a population-based cross-sectional survey conducted in the Netherlands of 7,200 people aged 65 years and older (with a response rate of 79%), 20% of the respondents were found to have nontraumatic foot complaints of more than 4 weeks' duration, often involving the forefoot. Female sex, joint disease, and multimorbidity were found to be risk factors for the presence of foot complaints; older age and obesity were not. Respondents with these complaints had limited mobility and poor perceived well-being. PMID- 11021052 TI - Outcomes research in podiatric medicine. AB - Outcomes research has become a high priority for the podiatric medical profession, according to the results of a recent survey of members of the American Podiatric Medical Association (APMA). The methods of population-based studies using claims data, health-related quality-of-life measures, decision and cost-effectiveness analysis, and quality improvement are discussed and their contributions to the podiatric medical profession highlighted. The integration of this methodology into the podiatric medical literature has been sparse. Future work needs to address the training of future researchers in these methods, the establishment of collaborative arrangements, and the development of uniform clinical and health-related quality-of-life measures. PMID- 11021053 TI - A case of peroneal neuropathy-induced footdrop. Correlated and compensatory lower extremity function. AB - This article reports on the case of a man with peroneal neuropathy-induced footdrop who was seen at the authors' institution 3 years after open reduction and internal fixation of a proximal fibular fracture and a distal, spiral, oblique tibial fracture of the right leg. A comprehensive gait analysis was conducted. A significant footdrop in gait resulted in a "reverse check mark" center-of-pressure pattern, an increased transverse-plane rotation of the foot, and excessive knee and hip flexion in the sagittal plane. These objective findings documented significant dysfunction within the involved lower extremity; in addition, aberrant biomechanics were observed in structures other than the site of initial injury within both limbs. PMID- 11021054 TI - Klippel-Trenaunay syndrome. An unusual presentation of unilateral leg edema. AB - Klippel-Trenaunay syndrome is a rare congenital condition that classically presents as a triad of varicosities, bone or soft-tissue hypertrophy, and cutaneous hemangiomas. The authors describe a case of this disease in an otherwise healthy patient with unilateral leg edema and intermittent bleeding from prominent ankle varicosities. The diagnosis was made on the basis of clinical presentation, diagnostic studies, and a comprehensive literature review. An overview of the pathology, treatment, and prognosis of this disease is provided. PMID- 11021055 TI - A rare occurrence of polydactyly. AB - Polydactyly is a common congenital foot deformity that varies in its clinical presentation. Duplication of the central ray accounts for 3% to 6% of all cases of pedal polydactyly. A review of polydactyly and its multiple classification schemes is presented, along with a rare case involving the duplication of a central digit and its surgical correction. PMID- 11021056 TI - [Recognizing malignant melanomas early enough. Saving the skin and life]. PMID- 11021057 TI - [Comprehensive skin cancer screening should be covered by national health insurance]. PMID- 11021058 TI - [The new EBM--a monstrosity? Colleague K. has missed the point]. PMID- 11021059 TI - [Increasingly more frequent refusal of prescription requests. Were physicians formerly too generous?]. PMID- 11021060 TI - [Turkish patient not accepted for the transplantation list. Scandalous prejudice toward foreigners?. Interview by Dr. Bernard Wiedemann]. PMID- 11021061 TI - [Patient with heart failure. Beta blockage becomes mandatory]. PMID- 11021062 TI - [Closer to a contraceptive drug for the man. How sperm are eliminated. Interview by Petra Eiden]. PMID- 11021063 TI - [Diarrhea and fever after vacation in the south. Specific to tropics only in every 5th patient]. AB - Post-travel medicine is mainly European medicine: Some 80% of the diseases are ubiquitous in nature, and only 20% tropics-specific. The leading symptoms are diarrhoea (68%), fever (20%) and skin conditions (9%), sometimes in combination. The travel history is of decisive importance for the diagnosis. Quite simple questions aimed at identifying the start of the illness, symptoms, destination, travel class and vaccinations can be useful for differentiation purposes. For the acute diagnosis, such aids as urine stick test, centrifuge, microscope, a range of stains, and a stool test for occult blood suffice. The most common serious travel-specific diseases include malaria, falciparum > tertian, dengue fever, hepatitis, and rickettsiosis. When falciparum malaria is suspected ("flu without a runny nose", and a visit to a tropical country), the patients must be hospitalized. Emergency diagnosis is done with a blood smear, "thick drop", and rapid malaria test. PMID- 11021064 TI - [Dangerous souvenir from Bali: scorpion in the luggage. First aid after sting--a case report]. PMID- 11021065 TI - [Barrett esophagus. Regular referral for endoscopy]. PMID- 11021066 TI - [Tea tree oil--household remedy of Australian aborigines. Complementary medicine methods, 5]. PMID- 11021067 TI - [Following blood specimen collection in patients with viral hepatitis: how dangerous is a needlestick injury?]. PMID- 11021068 TI - [Diabetic polyneuropathy. Sequential therapy with alpha-lipoic acid passes the general practice test]. PMID- 11021069 TI - [Diagnostic quiz. Panic attacks with surgical solution. Adrenal pheochromocytoma]. PMID- 11021070 TI - [High dosage acetylsalicylic acid for migraine. A classic drug with new appearance]. PMID- 11021071 TI - [Work site hypertension--an underestimated phenomenon. When job stress increases blood pressure]. PMID- 11021072 TI - [Interview with Prof. Joachim Schrader. Work site hypertensive patients are evidently more numerous than expected. Interview by Sonja Bohm]. PMID- 11021073 TI - [First insulin sensitizer approved in Germany. Insulin resistance in type 2 diabetes can be treated]. PMID- 11021074 TI - [Report of experiences. LDL alone is not always reliable enough]. PMID- 11021075 TI - Surgical strategy for meningioma extension into the optic canal. AB - Neuroimaging of the extension of meningioma into the optic canal was evaluated for planning the surgical strategy. Intracanalicular extension and localization were retrospectively analyzed in 13 patients with frontal base meningioma near the optic canal, based on the findings of visual field defects, magnetic resonance (MR) imaging, and surgical observations. MR imaging confirmed intracanalicular localization in one of three patients with tumors extending into the optic canals, and indicated the tumor in the others. The visual field defect did not precisely correspond to the tumor localization. Unroofing of the optic canal was performed in four patients and no adverse effects were observed. The interhemispheric approach was employed for tumors localized medially in the canal, and the pterional approach for tumors localized laterally. MR imaging is useful to evaluate the intracanalicular extension, but aggressive confirmation during surgery is essential. Tailored unroofing of the optic canal and removal of the intracanalicular tumor can be performed with few adverse effects and results in good tumor control. PMID- 11021076 TI - Immunohistochemical and ultrastructural study of clinically nonfunctioning pituitary adenomas. AB - Sixty-five clinically nonfunctioning pituitary adenomas were studied by immunohistochemistry, and 12 cases were also analyzed by electron microscopy. Thirty-nine cases (60%) were immunohistochemically identified as hormone producing adenomas. Six adenomas produced multiple hormones. Electron microscopy found seven null cell adenomas and five oncocytomas. The oncocytomas had a significantly higher incidence of hormone expression that the null cell adenomas. These results indicate that clinically nonfunctioning pituitary adenomas produce hormones, even though blood hormone levels are normal or low. Furthermore, the evidence of multihormonal production implies that two or more cell lineages including a protein hormone-producing type and a glycoprotein hormone-producing type may exist in the same nonfunctioning pituitary adenoma. PMID- 11021077 TI - Intracerebellar penetrating injury and abscess due to a wooden foreign body--case report. AB - A 4-year-old boy presented with chopstick penetration into the cerebellum via the temporal squamosa and tentorium cerebelli, which resulted in a cerebellar abscess 1.5 years after the injury. The neuroimaging appearance of the wooden chopstick were unusual, hyperdense on computed tomography, and isointense on T2-weighted and hypointense on T2-weighted magnetic resonance imaging. Abscess aspiration and continuous drainage was performed with real-time intraoperative ultrasound guidance. The chopstick fragment was surgically removed and the patient discharged with minor neurological deficits. Wooden foreign body may show changes in properties after a long period of intraparenchymal retention. Extra care is required to remove wooden foreign bodies because of the high risk of infection. PMID- 11021078 TI - Fatal severe vasospasm due to rewarming following hypothermia--case report. AB - A 37-year-old female died of cerebral vasospasm as a complication of rewarming following hypothermia therapy for severe head injury. She presented with severe consciousness disturbance and anisocoria after falling down a flight of stairs. Computed tomography (CT) revealed a right acute subdural hematoma and temporal contusion. Following surgery, mild hypothermia was started and rewarming was completed by the 11th day. Neurological examination showed no abnormalities, but intracranial pressure (ICP) suddenly increased and she manifested anisocoria on the 13th day. Repeat CT revealed a low density area in the right middle cerebral artery region and cerebral angiography showed diffuse narrowing of the main arterial trunks. A cerebrospinal fluid (CSF) sample was collected using an intraventricular ICP monitoring catheter. The CSF level of 8-hydroxy-2' deoxyguanosine was elevated during the rewarming period, indicating substantial deoxyribonucleic acid (DNA) oxidation. She died on the 15th day due to uncontrollable ICP. Histological examination at autopsy of the narrowed artery found the waving phenomenon in the internal elastic lamina and invasion of inflammatory cells into the adventitia. These findings constitute the possible evidence that free-radical-mediated oxidative DNA damage may be important in the genesis of severe vasospasm due to rewarming following hypothermia. PMID- 11021079 TI - Subepicranial varix mimicking sinus pericranii: usefulness of three-dimensional computed tomography angiography and bone window computed tomography--case report. AB - A 16-year-old female presented with a rare case of subepicranial varix in the left temporal area manifesting as a soft mass in the left temporal area when she laid down in the left lateral position. Bulging of the mass was observed when intracranial venous pressure was raised by the Valsalva maneuver, the left lateral position, or the prone position. Bone window computed tomography (CT) revealed a tiny hole, 1 mm in diameter, in the outer bone table. Three dimensional CT (3D-CT) angiography clearly visualized a mass with a diameter of approximately 10 mm connected to the diploic vein. The mass was totally resected by operation. Venous bleeding was observed from the tiny hole. Histological examination revealed a venous lesion mimicking sinus pericranii and containing endothelial cells. No communication with the intracranial venous sinuses was identified, so the diagnosis was subepicranial varix. Radiological examination by direct injection of contrast medium is usually performed to identify subepicranial varix, but 3D-CT angiography is a non-invasive preoperative examination that can visualize this small venous lesion. Adjustment of the CT acquisition conditions may allow 3D-CT angiography to identify sinus pericranii in the future. PMID- 11021080 TI - Giant aneurysm of the azygos anterior cerebral artery--case report. AB - A 77-year-old female presented with a giant aneurysm of the azygos anterior cerebral artery (ACA) manifesting as acute onset of akinetic mutism caused by enlargement of the aneurysm resulting from rapid thrombus formation within the aneurysmal sac. Thrombus removal to obtain decompression of the aneurysmal bulk and tension was performed before parent artery occlusion to prevent thromboembolic events. The aneurysmal neck was completely clipped with preservation of the parent artery and all branches. This strategy for direct neck clipping of a giant thrombosed distal ACA aneurysm can reduce the possibility of ischemic sequelae. PMID- 11021081 TI - Endovascular treatment of unusual multiple aneurysms of the internal carotid artery-posterior communicating artery complex--case report. AB - A 79-year-old female presented with subarachnoid hemorrhage due to rupture of a rare true posterior communicating artery (PCoA) aneurysm and with poor general condition. Endovascular therapy was performed in the chronic stage. Right carotid angiography just before embolization demonstrated unusual multiple aneurysms of the internal carotid artery (ICA)-PCoA complex. Superselective angiography and aneurysmography using microcatheter revealed two separate aneurysms arising from the PCoA and the ICA-PCoA junction. Endovacular embolization using Guglielmi detachable coils (GDCs) was successfully performed for both aneurysms and complete occlusions were achieved with the PCoA fully patent. Embolization with GDCs is a good alternative to surgical clipping for PCoA aneurysm after careful evaluation of superselective angiography. PMID- 11021082 TI - Terson syndrome caused by ventricular hemorrhage associated with moyamoya disease -case report. AB - A 24-year-old female presented with Terson syndrome secondary to bilateral ventricular hemorrhage as a complication of moyamoya disease. Ophthalmoscopy and magnetic resonance imaging clearly demonstrated vitreous hemorrhage in the left eye globe. Various visual symptoms are associated with moyamoya disease, almost all of which result from ischemic lesions in the visual cortex and optic pathways. In this case, the visual disturbance was caused by Terson syndrome secondary to ventricular hemorrhage. Close ophthalmological and radiological evaluation is mandatory even in patients with moyamoya disease and hemorrhagic manifestation located in the intracerebral, subarachnoid, or intraventricular space. PMID- 11021083 TI - Spontaneous intracranial hypotension associated with bilateral chronic subdural hematomas--case report. AB - A 34-year-old female presented with spontaneous intracranial hypotension (SIH) manifesting as severe postural headache and meningism. Magnetic resonance (MR) imaging with gadolinium showed diffuse pachymeningeal enhancement. She developed bilateral chronic subdural hematomas 4 weeks after the onset of the symptoms. MR imaging showed descent of the midline structures of the brain. The bilateral chronic subdural hematomas were surgically drained, with no remarkable pressure. Postoperative MR imaging showed complete resolution of the pachymeningeal enhancement and relevation of the midline structures of the brain. SIH is an uncommon and probably unrecognized condition because of the usually benign course. However, this case emphasizes that SIH is not entirely benign. SIH should be considered if there is no identifiable risk for intracranial hemorrhage, particularly in young patients. Neurosurgical intervention for the treatment of the underlying cerebrospinal fluid leak may be required if SIH persists. PMID- 11021084 TI - Hypoglossal neurinoma--two case reports. AB - Two patients presented with hypoglossal neurinoma extending both intra- and extracranially. A 63-year-old male presented with right trigeminal neuralgia and hypoglossal nerve paresis. The intracranial part of the tumor was removed totally via a suboccipital craniectomy. Over-coagulation of the venous collaterals, particularly the emissary veins, resulted in dural venous sinus thrombosis and cerebellar infarction. Unfortunately this patient died. A 48-year-old male presented with pareses of the VII, IX, X, XI, and XII cranial nerves and cerebellar sign. The tumor extended both extra- and intracranially, and was completely removed by opening the hypoglossal canal and the jugular foramen without over-coagulation of the venous collaterals. Preservation of the venous collaterals is very important for the prevention of postoperative venous complications. PMID- 11021085 TI - [A new mobile genetic element, staphylococcal cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus]. PMID- 11021086 TI - [Structure and functions of trypsin-like cysteine proteinases (gingipains) from Porphyromonas gingivalis, a causative bacterium of periodontitis]. PMID- 11021087 TI - [Biogenesis of lipoproteins in gram-negative bacteria: mechanisms underlying the selective membrane localization of lipoproteins]. PMID- 11021088 TI - [Modulation of host defense systems by beta-glucans]. PMID- 11021089 TI - [Progress in the diagnosis of bile duct stones]. PMID- 11021090 TI - [Treatment of common bile duct stones--from the viewpoint of physicians]. PMID- 11021091 TI - [Treatment of common bile duct stones--from the viewpoint of surgeons]. PMID- 11021092 TI - [A study on the appropriate time point for the assessment of Helicobacter pylori eradication--evaluation from the re-positive rate of H. pylori after successful eradication and delayed decrease of 13C-urea breath test levels]. AB - We attempted to evaluate the appropriate time point for the assessment of Helicobacter pylori eradication after treatment. One hundred and nine patients with gastroduodenal diseases were enrolled this study. All of them were received proton pomp inhibitor based triple therapy and diagnosed as eradication of H. pylori infection at initial assessment. They were followed up over six months. The diagnosis of H. pylori eradication was determined by rapid urease test, culture, histology and 13C-urea breath test (UBT), and the initial assessment of the eradication was performed on 31-90 days after finishing eradication therapy. Re-appearance rate of H. pylori after initial diagnosis of eradication was 4.6% (5/109), and the mean follow-up period of them was 16.3 months. The time period of initial assessment of eradication in these 5 patients were 35, 37, 42, 49 and 60 days after treatment, respectively. On the other hand, there were 6 patients who were diagnosed as failed of eradication therapy by 13C-UBT, and being success at following period. All of the 13C-UBT levels of these 6 patients were less than 10/1000 and were decreased within negative range subsequently. The time periods of initial diagnosis of these patients except one were within 2 months after treatment. It was concluded that the assessment time of H. pylori eradication should be performed over 2 months after eradication therapy. PMID- 11021093 TI - [A synchronous cancer of the esophagus and the trachea]. PMID- 11021094 TI - [A case of Cronkhite-Canada syndrome associated with colorectal cancer]. PMID- 11021095 TI - [A case of acute-onset autoimmune hepatitis with rheumatoid arthritis]. PMID- 11021096 TI - [An autopsy case of intrahepatic cholangiocarcinoma producing granulocyte-colony stimulating factor]. PMID- 11021097 TI - [A case of focal fatty infiltration of hepatic segment IV required to be distinguished from metastatic liver cancer]. PMID- 11021098 TI - [Clinical evaluation of the relationship between interferon therapy and oral lichen planus in patients with type C chronic hepatitis]. PMID- 11021099 TI - [Romanian ophthalmology]. PMID- 11021100 TI - [Diagnostic and treatment difficulties in patients with HIV and AIDS infections; the importance of interdisciplinary collaboration]. AB - At present, the immunodeficiency syndrome has became pandemic. In these conditions, every doctor, no matter his profile, will have to carry out patients with AIDS. This multisystemic disease goes beyond the limits of a single medical discipline, presenting a lot of clinical manifestations. In ophthalmology, the infection with immunodeficiency virus has a lot of clinical aspects. In the beginning stages, the ocular manifestations are nonspecific, often being the prime symptoms of the general infection. In time, the disease is advancing and the opportunistic infections are becoming very frequent. In the latest stages of the diseases, one can see ocular complications, recurrences of the retinitis and also accidents caused or related with therapy. We have formulated an algorithm of diagnosis, together with some elements of etiology, extremely important for the ocular determinations of the systemic disease. We have presented some general idea about the points, the aim and the principles of the antiretroviral treatment and about the monitoring activity of the disease. We have also presented an algorithm for the initiation of the treatment. As a conclusion, we notice the significance of a close cooperation between doctors, for the diagnostic and also for the therapeutical activity. PMID- 11021101 TI - [An update on the treatment of central retinal vein thrombosis (retinal vein occlusion)]. AB - The paper reviews the actual therapeutical means in a severe invalid eye disease. The treatment is not able to improve the retinal blood flow, being more effective on the main symptoms and complications. The isovolumetric hemodilution method as an up-to-date method in the treatment of central retinal vein occlusion, appears to bring better recuperation hopes, by a higher improvement of the local anatomical and hemodynamical factors. That is the reason why we shall expose it widely in our paper. Unfortunately, despite its complexity, the treatment of central retinal vein occlusion is not satisfactory from the point of view of the high number of patients that remain with a low visual acuity. The main goal of the therapy is the recovery of visual function, but a few aspects and especially the late presentation for ophthalmological examination, reduce the objective of treatment to prevent complications. PMID- 11021102 TI - [Specular microscopy--its clinical implications]. AB - Specular microscopy has multiple advantages in the clinical practice, allowing the study of the corneal endothelium in-vivo. We can observe corneal disorders, as well as the corneal status after intraocular surgery, of the posterior pole as well as of the anterior pole of the eye. PMID- 11021103 TI - [The neurogenic mechanisms of ocular inflammation and their modulation under local treatment with indomethacin]. AB - Neurogenic modulation of the inflammation is an essential mechanism of initiate and conditionate an inflammatory reaction. SP, NKA and NKB--neurokinins are involved in the modulation of the local ocular inflammation initiated by the surgical perforating trauma of the anterior pole. They determine the increase of vascular permeability and the breakdown of the aqueous-humor-blood barrier by direct action on the endothelial vascular receptors and by indirect mechanism (releases prostaglandins). This effect is demonstrated by the increase of the blue Evans concentration in the aqueous-humor after local application of SP, NKA and NKB in a dose-dependent manner. Local treatment with non-steroidal antiinflammatory drugs like Indomethacin stabilizes the aqueous-humor-blood barrier, by diminishing the local effect of neurokinins. By increasing the vascular permeability SP, NKA and NKB make an important linkage by which the nervous system mediators contribute to the modulation of eye inflammatory responses whose intimate mechanisms are not entirely known yet. PMID- 11021104 TI - [The hierarchization of the prognostic factors in uveal malignant melanoma]. AB - PURPOSE: To retrospectively study the prognostical factors, that influence disease development as well as survival in patients suffering of uveal malignant melanoma. METHODS: 100 cases of uveal malignant melanoma were thoroughly reviewed clinically and histopathologically. The analysis of the results were carried out by means of the statistically correlated methods. RESULTS: Factors having a negative prognostical influence upon the patient's survival were classified following their importance, the most significant of them being the histopathologically necrotic cellular type. CONCLUSIONS: Classifying the prognostically negative factors in patients with uveal malignant melanoma enabled to choose the most important of them in order to draw up a clinical record card suitable for ensuring a patient, whose eye had been enucleated due to an uveal malignant melanoma. PMID- 11021105 TI - [The congener eye in patients with unilateral classical malignant glaucoma]. AB - PURPOSE: To prospectively study in a follow-up period, ranging from 5 to 20 years, "the fate" of the fellow eye in patients with unilateral classical malignant glaucoma. METHODS: The fellow eye of 10 patients had been thoroughly examined immediately after the beginning of an adequate therapy of the unilateral classical malignant glaucoma. RESULTS: In 6 cases the fellow eye was apparently normal and 4 cases exhibited an intermittent primary angle-closure glaucoma. Surgery of the fellow eye was accepted immediately by two patients; in one a peripheral iridectomy was carried out and in the other one a trabeculectomy. The other 8 patients initially turned down surgery but accepted in thereafter in a period of time ranging between 1 and 8 years. A peripheral iridectomy was performed in two cases, a trabeculectomy in other two and a combined procedure (trabeculectomy with pars plana aspiration of the aqueous trapped within the vitreous in 3 cases and the same procedure plus lens extraction on one patient) in the remaining 4 cases, since they developed into a primary chronic angle closure glaucoma. CONCLUSIONS: Prophylactic surgery should be best done when the fellow eye is apparently normal having an entirely open angle and both the ocular tension and the visual function normal. The treatment of the advanced primary chronic angle-closure glaucoma of the fellow eye consists in trabeculectomy with pars plana aspiration associated with cataract extraction if the lens is opaque. PMID- 11021106 TI - [Changes in the vascular wall in the conjunctiva of patients with diabetes mellitus]. AB - Venous capillary wall in type II diabetic patients' conjunctiva was examined. The study was performed on conjunctiva fragments removed from diabetic patients during the operation of senile cataract. The fragments were fixed in Lillie's solution then studied by using optical microscopy with usual but histochemical stainings, too. Conjunctiva fragments removed from patients of the same age, being operated by senile cataract and one conjunctiva from ten years old patient were examined by using the same techniques as they could be compared. Computer determinations of the external and internal diameter of the same capillary in all three groups were performed. Venous capillary wall thickening by type IV collagen hyperproduction in all the aged patients was noted. This kind of thickening is more revealed in aged diabetic person. Type I-III collagen presence in the capillary wall of the diabetic patients was noted, too. The average value of the differential between the external and internal diameter was increased in the diabetic patients with diabetic retinopathy. Venous capillary wall thickening in the diabetic patients is due to the basal membrana both by means of the type IV collagen hyperproduction and presence of the I-III collagen, too. PMID- 11021107 TI - [Changes in the oscillatory potentials in the electroretinogram during the evolution of diabetic retinopathy]. AB - Oscillatory potentials where intensely studied in diabetic patients with various degrees of retinopathy. Classical reports pointed out the early reduction of their amplitude, as well as of their latency, while a and b waves remained still normal. A prognostic value was attached to these modifications, the reduction of oscillatory potentials being correlated with a ten time higher probability of developing a high risk retinopathy. 27 diabetic patients where investigated electroretinographically by us. No significant variation of the amplitude of oscillatory potentials was observed during the progression of diabetic retinopathy. PMID- 11021108 TI - [The significance of points alpha and beta on the dark adaptogram of the normal eye]. AB - PURPOSE: To determine the meaning of the points alpha and beta of the curve during dark adaptation. METHOD: In this paper I studied the curves during dark adaptation of the 48 normal eyes, examined at the Goldmann-Weekers adaptometer, for 30 minutes in standard conditions. The study had two stages: 1. To find out the mathematical low which defined the phenomenon; 2. To give a physiological explanation of the phenomenon. CONCLUSIONS: 1. Distribution in time of point alpha and point beta recognized mathematical low of normal log. 2. The cones adapted to the dark into two phases: from the beginning of the adaptation to the point alpha; after point alpha till before point beta. 3. The rods adapted in three phases: a) before point alpha to the point alpha; b) from point alpha to the point beta; c) after point beta to the terminal threshold. CONCLUSION: Point beta gained a new meaning: point beta represents the end of the cone's adaptation. PMID- 11021109 TI - [Ocular changes in dialysis patients]. AB - The study analyzes the ocular aspects in patients receiving hemodialysis, in order to define the importance of the ophthalmological exam as prognosis and follow-up parameter. The prospective study includes 84 patients with renal insufficiency who received hemodialysis between 1994-1998. The ocular aspects and their connection with the dialysis and the basic disease are described and analyzed. The most important were the retinal vascular complications: hypertensive retinopathy, anterior optic ischaemic neuropathy, central retinal artery occlusion, diabetic retinopathy. PMID- 11021111 TI - [The nonfunctional pituitary adenoma, The optic chiasm syndrome]. AB - We present the case of patient D. E., 48 years old, whose diagnosis is nonfunctional pituitary adenoma. The diagnosis was revealed by the presence of bitemporal hemianopia. This observation led us to remember the principal neuro ophthalmological manifestations and to emphasize the part of the ophthalmologist in the diagnosis of this disease. PMID- 11021110 TI - [An efficacy study of lodoxamide treatment in allergic eye lesions]. AB - Lodoxamid is an antiallergic drug, which stabilizes the mast cells' membrane blocking the release of the type I hypersensitivity reaction chemical mediators. A number of 25 patients with ocular allergic diseases (allergic conjunctivitis, vernal and atopic keratoconjunctivitis, giant papillary conjunctivitis), were included in this study. Lodoxamid, solution 0.1% (Alomide), was given 4 times daily for 6 weeks. The study's aim was to assess the lodoxamid's efficiency, on the ocular signs and symptoms. The study's results showed a significant improvement, or the disappearance of the ocular allergic disease. It is debated upon the lodoxamid's way and place of action, in blocking the type I hypersensitivity reaction. The lodoxamid's efficiency is due to its pharmacological features, by means of which it is effective on many links of the pathogenic chain: mast cells, eosinophils, lymphocytes, neutrophils, antigen presenting cells. Due to its action lodoxamid stabilizes the mast cell's membrane, and inhibits the release of histamine, prostaglandins, leukotrienes, triptase, interleukines -4, -8 and TNF-. During therapy with lodoxamid recruitment and activation of eosinophils is decreased, causing a significant reduction of the basic major protein, cationic eosinophilic protein, eosinophilic derived neurotoxin, eosinophilic peroxidase. Lodoxamid reduces the expression of ICAM-1 on the surface of the antigen presenting cells, and decreases the number of the TH2 cells, from the tears of the allergic patients. PMID- 11021112 TI - [Orbital mesenchymal tumors--orbital fibrous histiocytoma. Their clinical and histopathological aspects]. AB - Histiocytoma is a tumor with mesenchymal etiology, whose primary orbital form is very rare. (1) Although it is a benign tumor, there have been written--in a few cases--it's metastatic forms too. This article describes the case of a female patient with this tumor which had been extirpated successfully and without any recurrences till now. The disease had been appeared with a chronic sphenoidal and maxillary sinusitis. At the same time there are described the latest concepts about these tumors' etiology, classification and their treatment. PMID- 11021113 TI - [Ocular cicatricial pemphigoid]. AB - Ocular cicatricial pemphigoid is a systemic disease of autoimmune etiology, belonging to the category of acquired oculo-muco-cutaneous bullous dermatosis. Although it is a disease primarily involving the conjunctiva, it frequently affects other mucous membranes (oropharynx, genitalia, anus), and the skin is involved as well in approximately 15% of the cases. The ocular manifestations, during the acute stage, consist of conjunctival bullous, which quickly lead to ulcerous lesions. During the chronic stage the disturbances of the palpebral statics and dynamics, ocular dryness syndrome and corneal lesions will develop. Being a relative less frequent disease, the observation of 3 patients is presented. PMID- 11021114 TI - [A familial case of drusen of the optic disk]. AB - The present abstract is about a case of drusen of the optic disc (DOD), consisting of three members, belonging to two generations: the mother (41 years) and two daughters (16 and 19 years). Their personal records showed nothing special, except case III-2 with crisis equivalent to the epileptic ones. No changes of the visual acuity and of the visual field were found. Ophthalmoscopy and angiofluorography showed: a bilateral and superficial location of the DOD; no vascular anomalies (discal and peridiscal) or other retinal changes; a trend towards flattening and getting more mat of the DOD, in older cases. It is debated upon the possibility that a single gene might be involved, responsible for an autosomal dominant or autosomal recessive transmission mode. PMID- 11021115 TI - [Young ophthalmologists]. PMID- 11021116 TI - [The mechanisms of neuronal death in glaucoma]. AB - The current therapy of glaucoma (drugs, LASER, surgery) acts mainly on the trabecular meshwork and on the ciliary body. The loss of the visual function in glaucoma is however determined by the disfunction and death of the ganglion cells. The main theories trying to explain the death of the ganglion cells in glaucoma are analyzed. Their practical value is represented by the finding of the ideal neuroprotective drug. PMID- 11021118 TI - [The explantation and implantation of artificial crystalline lenses]. AB - THE PURPOSE of the study is to evaluate the causes of explantation and replacement of different types of intraocular lenses. MATERIAL AND METHOD: We performed a study of 1060 eyes operated for cataract in the University Eye Hospital between 1992-1996, following the number of explantation and replacement of intraocular lenses (IOL), the types of IOL and the causes of explantation. RESULTS: We performed 8 explantation of IOL (0.75 g) for pseudophakic bullous keratopathy (1 case), chronic uveitis (3 cases), secondary glaucoma (2 cases), suppurative uveitis (1 case), cystoid macular oedema (1 case). From the 8 explantation of IOL, 3 were anterior chamber intraocular lenses (AC-IOL) and 5 posterior chamber intraocular lenses (PC-IOL). We performed reposition in 4 cases of IOL. The most frequent cause of explantation was chronic uveitis and secondary glaucoma. CONCLUSION: Explantation is indicated in certain cases of IOL complications without response on medical treatment. PMID- 11021117 TI - [Modern concepts in the pathogenesis and treatment of glaucoma]. AB - The reduction of the visual performances in glaucoma may be explained by several theories. The most admitted theory, for the present, considers glaucoma as a progressive primitive optic neuropathy, having the origin at the retinal ganglion cells, associated with a disease of the sclero-corneal trabeculum, whose porosity is diminishing. The most important pathogenic hypothesis in glaucoma are: the vascular, the mechanical and the collagenic hypothesis. The vascular theory considers that the prolonged ischemia is a responsible for the neuronal alteration. In this process are participating a lot of factors: the intraocular pressure, the blood pressure and some other factors governing the vascular tone of the optic nerve (myogenic factors, metabolic factors, vasoactive factors, circulating hormones and endothelial factors). From therapeutical point of vue, these modern concepts are reflected in the new therapeutic perspectives, with a special attention for the axonal outflow and trabecular cell disfunction and also for the dysregulation of the cellular oxidoreduction. PMID- 11021119 TI - [Chorioretinal lesions in deaf mutes]. AB - The paper reports a study of 216 patients of special deaf-mute schools from Craiova. The exam of them showed us the presence of retinal degenerative lesions in 17 cases (7.8%). There was not a parallelism between deafness' degree and retinal lesions expanse. In point of clinical aspect, the retinal degenerative lesions were Sjogreen retinal lesions in 6 cases, colloidal Amalric-Bessou retinal lesions in 6 cases. In 2 cases lesions had a pseudoinflammatory aspect and another 2 cases were peripheral lesions like "salt and pepper's". Only one case had pigmentary osteoblastic mobilization. Deafness was associated with oligophrenia in 15 cases, with discreet manifestations of eredoataxia Friedreich in 5 cases, with epilepsy in 2 cases and with nanism in another 2 cases. It is necessary a complex oto-neuro-ophthalmological exploration of patients with deaf muteness. PMID- 11021120 TI - [The effects of photocoagulation on the electrical activity of the diabetic retina]. AB - The aim of photocoagulation in diabetic retinopathy is to destroy ischaemic retina because this produces a vasogenic factor, responsible of neovascularization. Retinal burn reduces the number of photoreceptors and effects the transmission of nervous impulse in areas close to those coagulated. It is presumable therefore that electroretinographic parameters will be significantly modified after treatment. 23 diabetic patients where investigated, resulting 43 scotopic electroretinograms, 41 photopic electroretinograms and 46 flicker electroretinograms, each eye being considered separately. The statistic analysis showed significant alterations with laser treatment of the amplitudes of a and b waves in both scotopic and photopic conditions as well as diminished amplitude of the flicker test. PMID- 11021121 TI - [Changes in myopic refractive errors after 9 months of extensive wear of hydrogel lenses with high oxygen permeability and compared with those with low permeability]. AB - BACKGROUND: A small but significant increase in myopia after extended wear of low oxygen permeability (Dk) hydrogel lenses has been previously reported; however, the specific impact of hypoxia on refractive status and corneal curvature with extended wear are not well documented. THE PURPOSE of this study was to compare the refractive changes induced over a period of 9 months' extended wear with high Dk fluorosiloxane hydrogel lenses and low-Dk hydrogel lenses. METHODS: Adapted daily wear contact lens wearers were randomly assigned to one of two groups. The low-Dk group wore etafilcon A (Dk = 28) for up to 7 days and 6 nights and the high-Dk group wore lotrafilcon A (Dk = 140) for up to 30 days and nights. Refractive error and corneal curvature were measured at 3-month intervals over 9 month of extended wear. RESULTS: The etafilcon A group demonstrated an average increase in myopia of 0.30 D over the 9 months period; however, no change in spherical myopic correction was measured in the lotrafilcon A group. The cylindrical component did not change in either group. A stratified analysis revealed a greater increase in myopia for low myopes than moderate myopes in the etafilcon A group but no difference in the lotrafilcon A group. Keratometric analysis revealed no change in the etafilcon A group and a small degree of central corneal flattering in both major meridians of 0.35 D in the lotrafilcon A group. CONCLUSIONS: Nine months of extended wear of low-Dk lenses is associated with a small degree of myopic progression in adult myopes that appears to be reversible. Wearing fluorosiloxane-hydrogel lenses of high-Dk had no impact on refractive error and may be associated with a small degree of central corneal flattering. PMID- 11021122 TI - [The clinical aspects in myopic retinal detachment]. AB - Beside aphakia and senility myopia is one of the major factors predisposing to retinal detachment. The purpose of the study is to define the clinical features of the myopic retinal detachment with therapeutic and prognostic implications. METHOD: 100 myopic (< 6 dioptres) retinal detachments that were hospitalized in the Ophthalmological Department from Cluj-Napoca between 1996-99 are retrospectively analyzed. RESULTS, DISCUSSION: The major clinical elements having an impact in defining the anatomical and functional prognosis of the myopic retinal detachment are described and analyzed: the extension of the retinal detachment, the number and shape of the retinal ruptures, the detachment of the macula, the association of aphakia and of myopic choroidosis. PMID- 11021123 TI - [A survival analysis of patients with malignant uveal melanoma]. AB - PURPOSE: Survival rate estimate in patients with uveal malignant melanoma. METHODS: 100 cases of uveal malignant melanoma were retrospectively reviewed clinically and histopathologically. Survival rate assessment of the patients was carried out by means of uni and multivariate analysis. Univariate evaluation was performed using the Kaplan-Meier's method and multivariate analysis by means of the proportional hazards model of Cox. RESULTS: Univariate analysis disclosed some significant factors predicting survival rates such as: the period of time elapsed between occurring the first disease symptoms and the objective appearance of uveal melanoma, personal history connected with melanoma, the size, shape and extrascleral extension of the tumor, the Callender's cell type, the reticulin and melanin amount existing within the tumor, the necrosis extension into the tumor and development or not of the metastases. Multivariate analysis succeeded in disclosing a hierarchy of factors predicting survival rates headed by the histopathologically necrosis cell type. CONCLUSIONS: There are not significant differences between sexes, different age groups and professions with regard to survival rates of the patients with uveal malignant melanoma. Multivariate analysis represents the main means of assessment of the part played by a combination of different prognostical factors in survival of uveal malignant melanoma patients. PMID- 11021124 TI - [Colobomatous pits of the optic nerve papilla associated with serous retinal detachment. The clinical and pathogenic aspects]. AB - The present study is about a number of 19 cases (19 eyes), the patients having a congenital pit of the optic papilla (CPP) complicated with serous retinal detachment (SRD). The cases were selected, from a group of 41 patients with CPP. Average age of the cases was 37(31-54) years. The serous retinal detachment appeared under two different clinical appearances: the detachment of the internal retinal layers (DIRL) or retinal schisis, 10 cases, located between the papillae and macula, and involving the optic disc; the detachment of the external retinal layers (DERL), 9 cases, located within the macular area, without involvement of the optic disc. A number of 8 cases (42%) developed a macular hole, located within the external retinal layers. The cases with DIRL had better visual acuity, compared to those with DERL. This study proves the bilamellate character, of the serous retinal detachment. The disease starts with DIRL (retinal schisis), due to fluid passing at the level of the CPP and then DERL develops as a complication, following the degenerescence of the retinal layers within this area. PMID- 11021125 TI - [The incidence of ocular complications in 10,000 diabetes mellitus patients hospitalized at Dolj County Clinical Hospital]. AB - The paper surveys the statistical results of retrospective analysis for observation sheets of patients having mellitus diabetes, type I and II, hospitalized during the last 10 years at the Ophthalmological Clinic in Craiova. The accent was made on the ocular complications related to the type of mellitus diabetes, the age of its debut, the evolution length, the treatment made, glycemia compensation, which make this affection to be the first cause of cecity at adult age. PMID- 11021126 TI - [Isovolemic hemodilution--a treatment method for central retinal vein obstruction]. AB - OVCR--is a vascular disease of the posterior pole of the eye, witch leads usually, to a significant decrease of the visual acuity. The actual management of the disease is still inefficient of the functional recovery of those patients. Among the medical treatment methods, is also the isovolemic hemodilution, witch must be established only after a thorough cardio-vascular and renal examination. The present study shows an increase of the visual acuity in the patients treated with isovolemic hemodilution followed by LASER Argon photocoagulation. PMID- 11021127 TI - [Allergic retinal vasculitis following the intramuscular administration of Moldamin]. AB - The authors presents two cases report of allergic retinal vasculitis and allergic optic neuropathy after anaphylactic shock due to the parenteral Moldamin administration. Clinicians should be aware of the possibilities of hypersensitivity ocular reaction in anaphylactic shock, with dramatic consequence on the vision. PMID- 11021128 TI - [Retinoschisis--the clinical associations and therapeutic possibilities]. AB - Retinoschisis is a vitreo retinal dystrophy characterized by splitting of the neuroretinal layers. The paper presents two cases of retinoschisis associated with retinal detachment. As a particularity, one patient was diagnosed with von Willebrand disease and the other one, with otomandibular dysostosis (Francois Haustrade). There are presented therapeutical methods used in this two cases: Laser, Argon photocoagulation, transscleral cryopexia, scleral buckling and also the functional and anatomical results. PMID- 11021129 TI - [Changes to visual acuity and the visual field in a case of meningioma operated at the anterior level]. AB - It's presented a midline meningioma of anterior level case which preoperative has an importance affectation of the visual acuity and visual field, and postoperative in dynamic is founded the important improvement of visual acuity and visual field. It's discussed visual field topography and atrophy optic pathophysiology. It's a neuro-ophthalmology case example, where the cooperation neurosurgery-ophthalmology permit medical solution and restored in social life of the patient. PMID- 11021130 TI - [The growth and decline of medical publications]. PMID- 11021131 TI - [Modern methods for examining the cornea]. AB - The paper makes a review of indications limits and possibilities of some unusual diagnostic techniques: pachymetry; ultrasound biomicroscopy and confocal microscopy. PMID- 11021132 TI - [Optical coherence tomography]. AB - Optical coherence tomography is a novel, noninvasive, noncontact and repetitive imaging technique which produces high depth resolution cross-sectional tomographs of ocular tissue. It's value is given by the possibility of achieving pseudo histological images of the target tissue. It is optically based, analogue to ultrasound B-scan examination and similar to laser reflectometry. Optical coherence tomography involves shining low-level infrared light on a tissue specimen, an interferometer and a computerized imaging system. The most attractive applications of optical coherence tomography are the diseases of the retina such as: central serous retinopathy, macular hole, chronic macular edema, age-related macular degenerescence. PMID- 11021133 TI - [The influence of meteorological factors in wintertime on the incidence of the occurrence of acute endogenous iridocyclitis]. AB - PURPOSE: I tried to establish if the atmospheric factors such as: temperature, pressure, humidity of the air, wind, had any influence on the onset of acute uveitis. METHOD: This study was made on 597 cases with acute uveitis hospitalized in the Eye Clinic--Cluj-Napoca, between 1983-1991. CONCLUSION: During winter, in the region of Cluj, acute uveitis occurred significantly more often: 1. When the weather was warmer or colder than the mean value of the season, and the temperature varied with more than 4 degrees C from the previous day; 2. When the humidity of the air was low (dry weather); 3. When the wind was more than 4 m/s. Correlating the physiological knowledge with the studies about weather and the living organisms, I draw some possible pathogenetic mechanisms to explain the influence of the meteorological factors on the onset of acute uveitis, in winter. PMID- 11021134 TI - [The incidence of primary open-angle glaucoma in patients with central retinal vein occlusion]. AB - PURPOSE: To prospectively study in a 10-year follow-up period the dynamics of the incidence of the primary open-angle glaucoma in patients with central retinal vein occlusion. METHOD: The study group consisted of 147 patients without diabetes with open angle glaucoma remained constantly by the end of the 10th month of the follow-up period when the frequency dropped to 4.7% due to the occurrence of neovascular glaucoma in one case of venous occlusion associated with primary open-angle glaucoma. The progressively increasing incidence of primary open-angle glaucoma to 10.2% thereafter was accounted for by the fact that in 8 patients the glaucoma suspect associated with central retinal vein occlusion converted to primary open-angle glaucoma. The incidence of primary open open angle glaucoma in patients of the control group was rather constant, ranging between 4 and 5%. CONCLUSIONS: There was no difference in the first years of the follow-up between the incidences of the primary open-angle glaucoma in both the study and control group. The obvious difference between the two frequencies appeared after the third year and by the end of the ensuing the two frequencies of primary open-angle glaucoma doubled in the patients of the study group. The low incidence of the neovascular glaucoma (1/16 = 5.5%) showed that the primary open-angle glaucoma associated with central retinal vein occlusion represented a protecting factor preventing the occurrence of the neovascular glaucoma. PMID- 11021135 TI - [The clinico-statistical correlations of eye injuries in children]. AB - This is a study of epidemiological and medico-social aspects of eye injuries in children. It refers to 624 cases, over the period 1996-1999; only 108 of them needed admission. The analysed factors are: the alleged person, the traumatic agent, the socio-climatic environment and the medical aspects of the accident. Only mechanical traumas are considered. The study reveals the quantitative indicators of various aspects such as: medical attendance, surgery, drug treatment, decreases of vision. PMID- 11021136 TI - [Inflammatory optic neuropathies]. AB - The article presents a study on 94 patients hospitalized in the Clinic of Ophthalmology of Cluj-Napoca, during 1993-1998, with optic neuritis. There are presented the clinical and functional aspects of optic neuritis, especially the importance of static perimetry, for the diagnosis, and her superiority, compared with the dynamic perimetry, in finding minor visual field defects in the diagnosis of the optic neuritis too. After treatment, 75% of the cases have a favorable evolution, but the static perimetry reveals sequela visual field defects at 85.3% of the eyes which suffered optic neuritis. According with this findings, static perimetry has a important role in the diagnosis of optic neuritis, in the revealing the functional sequela defects and in the monitoring of the patients. PMID- 11021137 TI - [Anterior ischemic optic neuropathies]. AB - The article presents a study on 82 patients hospitalized in the Clinic of Ophthalmology of Cluj-Napoca, during 1993-1999, with anterior ischaemic optic neuropathy (AION). There are presented the clinical and functional aspects of AION, especially the importance of static perimetry (PS) for the diagnosis, and her superiority, compared with the dynamic perimetry, in the monitoring of the patients with chronic nerve head ischaemia. After treatment, 32.9% of the cases have a favorable evolution, but PS reveals sequela visual field defects at all eyes which suffered AION. Long-term monitoring of the patients reveals unfavorable evolution at mare than 85% of cases, because of the chronic residual ischaemia, characterised by decreasing of visual capacity in PS. According with this findings, static perimetry has a important role in the diagnosis of AION, in revealing of the sequela defects and in the monitoring of the patients. PMID- 11021138 TI - [Postcontusion glaucoma]. AB - Secondary posttraumatic glaucoma. Blunt ocular trauma may cause transient or permanent high IOP, occurring immediately or in time after the injury. This study presents some clinical and therapeutical considerations about 16 cases which developed secondary glaucoma. The elevation of IOP was associated with hyphema, iridodialysis, zonular tears (with lens luxation or subluxations), alterations of the iridocorneal angle. PMID- 11021139 TI - [A myopia study of glaucoma patients following pilocarpine instillation]. AB - AIM: Our research studies transitory myopia induced by Pilocarpine treatment in glaucoma patients. MATERIAL AND METHODS: 48 patients with open angle glaucoma were selected, visual acuity and refraction were measured before and after the instillation of Pilocarpine 2%, for two hours. Induced myopia was compared for different ages and it was established the influence of Timolol and Acetazolamide administered in the same time. RESULTS: Transitory myopia is important in young patients, reduced after 50 years and absent after 70 years. Acetazolamide increases myopia, Timolol has no effect on refraction. CONCLUSIONS: The study confirms the unpleasant effect of Pilocarpine and suggests not performing refractometry in the first two hours after the instillation of the drug. PMID- 11021140 TI - [Anterior ischemic optic neuropathy secondary to retrobulbar hematoma]. AB - Two clinical cases are displayed, age over 70, with multiple vascular risk factors; following the retrobulbar injection performed for cataract surgery, retrobulbar hematoma and ischemic anterior neuropathy developed as complications. Pathogenesis of the optic ischemic anterior neuropathy may be explained by changes of a precarious hemodynamic balance and witch was previously settled, due to mechanic compression and increase of the intraocular pressure, which lead to the decrease of the blood pressure and implicitly to the development of optic neuropathy. PMID- 11021141 TI - [Isolated obliteration of the cilioretinal artery]. AB - Place on record the case patient of 51 years, who evince diminution visual acuity to the right eye, due to one precapillary occlusion of cilioretinal artery. Clinical syndrome to be characterized through diminution visual acuity, ophthalmoscopic (ischemic edema) and angio-fluorographic alters (the place of obliteration, ischemic zone, coterminous vessels alters, the absence of soft exudates). Etiopathogenic to be discussed the possibility one cholesterolemic ambole due to its localization and appearance ophthalmoscopic as well easily value of cholesterolemic. Functional prognostic was has been, due to localization obstruction of circulation preterminal, unique character of cilioretinal artery, as well anatomical specific features vascularity from the level of macula. PMID- 11021142 TI - [Suprainfected corneal injuries--the specific therapeutic aspects in 2 cases]. AB - This paper presents two cases of corneal abscess and their therapeutically approach which was the surgical treatment (penetrant keratoplasty). First case was a corneal ulcer which failed to abscess under medical therapy. The second case was a corneal abscess complicated with impending perforation after intracorneal foreign body. The particular aspects for each case are emphasized as well as the common aspects for corneal infections summarized as an algorithm. PMID- 11021143 TI - [Serpiginous choroiditis--the diagnostic problems]. AB - Serpiginous choroiditis is a very rare ocular disease with an unknown etiology. This paper presents the case of a patient with serpiginous choroiditis and the diagnosis problems of this case. PMID- 11021145 TI - Introduction to the special section on methods and implications of revising assessment instruments. PMID- 11021144 TI - [An inclusion cyst of the anterior chamber]. AB - Our paper present a clinical case of inclusion cyst of the anterior chamber. We discuss the clinical course, the specific investigations, the surgical management and the histopathologic diagnosis. PMID- 11021146 TI - Implications of test revisions for research. AB - Test revisions are increasingly common in psychology and neuropsychology in particular. However, such revisions may alter in complex ways the kind of information obtained, and they may assess traits, abilities, and conditions in ways different from earlier versions. This article outlines some of the problems associated with the revision of tests facing clinicians and researchers. Three broad classes of revision are considered. Part 1 considers the aging of tests, part 2 concerns the aging of participants, and part 3 considers changes in test format. Although the article focuses largely on measures of intelligence and personality, the issues addressed in the article apply to other tests and assessment domains as well. PMID- 11021147 TI - Does the rose still smell as sweet? Item variability across test forms and revisions. AB - This article examines item stability when the same item appears in different contexts. The 1st section considers the assumptions in classical test theory and item response theory concerning the relationship between the item and the trait it is presumed to measure. The 2nd section presents contextualist challenges to the measurement theory assumptions about item properties and shows the instability of item characteristics across different testing contexts. The 3rd section describes methods for checking the relationship between items and traits. Classical test methods, item response methods, and structural equation methods for assessing item stability are reviewed. The instability of item characteristics across contexts should caution researchers to assess, and not assume, that items operate the same way on different test versions. Item instability also indicates the need for a more detailed understanding of the psychological processes that occur between item and answer. PMID- 11021148 TI - Updating to the WAIS-III and WMS-III: considerations for research and clinical practice. AB - The Wechsler Adult Intelligence Scale-Revised (WAIS-R) and the Wechsler Memory Scale-Revised (WMS-R) are the most commonly used intelligence and memory scales in both clinical and neuropsychology. In 1997, updated versions of these instruments (the WAIS-III and WMS-III) were published. Because of the extensive use of the WAIS-R and WMS-R in the field and the body of accumulated research, there is naturally some reluctance by clinicians and researchers to update to the new versions. It is sometimes difficult for clinicians who test individuals on repeated occasions to switch over to the new versions of the scales because of the difficulty of interpreting score discrepancy between the 2 versions. Researchers, especially those conducting longitudinal research, have a similar difficulty in changing measurement devices because of the possible threat of internal validity. This article reviews the substantive revisions of the scales and outlines those issues that users should take into consideration when updating to the new versions. PMID- 11021149 TI - Revising psychological tests: lessons learned from the revision of the MMPI. AB - Some types of psychological tests become dated and require more frequent and more extensive revision than others. Because of the formidable effort that is required in a test revision, the goals and scope of the revision need to be carefully staked out before a revision is undertaken. The revision team needs to develop a generally agreed-upon guiding philosophy for the test revision in the beginning of the project and incorporate broad input into the changes that are likely to be required. Factors important to consider in a test revision are discussed, and the parameters of personality test revision illustrated from the extensive program to revise the Minnesota Multiphasic Personality Inventory (MMPI) are included. Recommendations for gauging acceptance of the revision are suggested along with steps that revisers and publishers might take to make a test revision both more research based and more acceptable to test users. PMID- 11021150 TI - Implications of test revisions for assessment with Asian Americans. AB - There are serious gaps in knowledge with respect to the use of standardized assessment instruments such as the Wechsler Adult Intelligence Scale-Third Edition (WAIS-III; D. Wechsler, 1997) or the Minnesota Multiphasic Personality Inventory-2 (MMPI-2; J. N. Butcher, W. G. Dahlstrom, J. R. Graham, A. Tellegen, & B. Kaemmer, 1989) with Asian Americans. Issues surrounding the availability, reliability, and validity of assessment instruments must be addressed before extended discussions about the implication of test revisions for this population can take place. The authors review the current status of the WAIS-III and MMPI-2 with Asian Americans with respect to their availability, reliability, and validity, including reasons why Asian Americans have been severely underrepresented in validation studies. The authors argue for the need to collect data on the use of standardized assessment instruments with Asian Americans and conclude with recommendations for the inclusion of this population in future test revision projects. PMID- 11021151 TI - Practical and ethical issues pertaining to test revisions. AB - Revisions of clinical psychological tests are occurring at more frequent intervals than in the past. These revisions involve such practical issues as the goals of the revision, the economic considerations relating to the revision, the methodology of the revision, and the degree to which the revision meets the available scientific and ethical standards governing the use of these tests in individual assessment. A tension between practical and ethical issues in the test revision process is inevitable and demands the best of psychologists in their decision making. Test developers, psychologists, patients, and consumers of the test interpretations and recommendations have legitimate interests and a stake in seeing that test revisions are applied with maximal effectiveness and fairness in the broadest sense. PMID- 11021152 TI - Factor analysis and scale revision. AB - This article reviews methodological issues that arise in the application of exploratory factor analysis (EFA) to scale revision and refinement. The authors begin by discussing how the appropriate use of EFA in scale revision is influenced by both the hierarchical nature of psychological constructs and the motivations underlying the revision. Then they specifically address (a) important issues that arise prior to data collection (e.g., selecting an appropriate sample), (b) technical aspects of factor analysis (e.g., determining the number of factors to retain), and (c) procedures used to evaluate the outcome of the scale revision (e.g., determining whether the new measure functions equivalently for different populations). PMID- 11021153 TI - Clinical and research implications of revising psychological tests. AB - This article integrates those of other contributors to this special section, "Methods and Implications of Revising Assessment Instruments," to underscore important conceptual factors to consider when undertaking test revisions. These considerations include determination of when test measures have become sufficiently understood to be incorporated in a test revision, cohort effects, revision of administration formats and test instructions, and comparisons of performance levels across test versions. The discussion of these factors also takes into consideration clinical practice and educational implications of making a transition to revised test versions. PMID- 11021154 TI - Child-Adolescent Suicidal Potential Index (CASPI): a screen for risk for early onset suicidal behavior. AB - This study's purpose was to develop a reliable and valid self-report questionnaire, the Child-Adolescent Suicidal Potential Index (CASPI), to screen for risk for suicidal behavior in children and adolescents. Four hundred twenty five child and adolescent psychiatric patients and nonpatients completed the CASPI and other research instruments to rate suicidal and assaultive behavior and symptoms of depression, anxiety, and hopelessness. The 30-item CASPI involves 3 factors (anxious-impulsive depression, suicidal ideation or acts, family distress) that contributed to a unidimensional 2nd-order factor accounting for 59% of the total variance. Internal consistency (alpha) for the total score was .90, and test-retest reliability (ICC) for the total score was .76. Total score distinguished between children and adolescents with different severity of psychopathology and different levels of suicidal and assaultive behavior. Each of the 3 factors had different contributions to discriminating between levels of suicidal status. CASPI total score of 11 distinguished suicidal ideation or acts from nonsuicidal behavior, with sensitivity 70% and specificity 65%. CASPI total score positively correlated with symptom severity of depression, anxiety, and hopelessness. PMID- 11021155 TI - The discriminative validity of a phallometric test for pedophilic interests among adolescent sex offenders against children. AB - The authors examined the responses of adolescent sex offenders against children on a phallometric test of pedophilic interests. Participants were 40 adolescent sex offenders against children, 75 young adult sex offenders against children, and 39 young adult comparison participants. The responses of adolescents with female victims resembled those of comparison participants; adolescents with any male victims had larger relative responses to child stimuli than comparison participants. Young adult offenders, regardless of victim sex, had larger relative responses to child stimuli than comparison participants. Using a cut score of 0 (indicating equal or greater arousal to children than to adults), sensitivity was 42% for adolescents with any male victims, and specificity was 92% for the comparison participants. Results suggest phallometric testing can identify pedophilic interests among these adolescent sex offenders. PMID- 11021156 TI - Comparison of white, African American, Hispanic, and Asian children on the Naglieri Nonverbal Ability Test. AB - This study examined differences between 3 matched samples of White (n = 2,306) and African American (n = 2,306), White (n = 1,176) and Hispanic (n = 1,176), and White (n = 466) and Asian (n = 466) children on the Naglieri Nonverbal Ability Test (NNAT; J. A. Naglieri, 1997a). The groups were selected from 22,620 children included in the NNAT standardization sample and matched on geographic region, socioeconomic status, ethnicity, and type of school setting (public or private). There was only a small difference between the NNAT scores for the White and African American samples (d ratio = .25) and minimal differences between the White and Hispanic (d ratio = .17) and between the White and Asian (d ratio = .02) groups. The NNAT was moderately correlated with achievement for the total sample and correlated similarly with achievement for the White and ethnic minority groups. The median correlation of NNAT with reading was .52 and NNAT with math was .63 across the samples. Results suggest that the NNAT scores have use for fair assessment of White and minority children. PMID- 11021157 TI - Normative data for the F(p) scale of the MMPI-2: implications for clinical and forensic assessment of malingering. AB - Data from several clinical samples and the Minnesota Multiphasic Personality Inventory--2 standardization group are presented to familiarize the reader with response patterns of different groups on a new validity scale designed to assist in the identification of exaggeration or fabrication of psychological disturbance. Sensitivity-specificity analyses are included along with suggestions for use of the F(p) Scale with other validity scales. Cautions about setting single cutoff scores are also discussed. PMID- 11021158 TI - Confirmatory factor analyses of the WAIS-III standardization data. AB - Confirmatory factor analyses with the standardization data of the Wechsler Adult Intelligence Scale-Third Edition (Wechsler, 1997a) compared 6 models with 1 to 4 factors for 11- and 13-subtest versions of the test. Three factors usually fit the data better than 2 factors, but 2-factor models were more parsimonious. A 2 factor model with a Verbal Comprehension factor (Vocabulary, Similarities, Information, and Comprehension) was as good as and sometimes better than the 2 factor model defined by the traditional separation of Verbal and Performance subtests. For 3-factor models, alternative specifications of processing speed subtests on either the Perceptual Organization or Freedom From Distractibility factor were comparable, and specifying a 4th factor for Digit Symbol and Symbol Search had little advantage in comparison with 3-factor models with correlated errors for the 2 subtests. PMID- 11021160 TI - Summed-score linking using item response theory: application to depression measurement. AB - An item response theory (IRT) approach to test linking based on summed scores is presented and demonstrated by calibrating a modified 23-item version of the Center for Epidemiologic Studies Depression Scale (CES-D) to the standard 20-item CES-D. Data are from the Depression Patient Outcomes Research Team, II, which used a modified CES-D to measure risk for depression. Responses (N = 1,120) to items on both the original and modified versions were calibrated simultaneously using F. Samejima's (1969, 1997) graded IRT model. The 2 scales were linked on the basis of derived summed-score-to-IRT-score translation tables. The established cut score of 16 on the standard CES-D corresponded most closely to a summed score of 20 on the modified version. The IRT summed-score approach to test linking is a straightforward, valid, and practical method that can be applied in a variety of situations. PMID- 11021159 TI - An empirical investigation of the factor structure of the AUDIT. AB - This study investigated the Alcohol Use Disorders Identification Test's (AUDIT) factor structure and psychometric properties. The factor structure was derived from a sample of 7,035 men and women primary care patients. A principal components analysis identified 2 factors in the AUDIT data and was supported in a confirmatory factor analysis (CFA). The 2 factors were Dependence/Consequences and Alcohol Consumption. The CFA also provided support for a 3-factor model whose factors (Alcohol Consumption, Alcohol Dependence, and Related Consequences) matched those proposed by the AUDIT's developers. Psychometric indexes were determined by use of the baseline and 12-month follow-up data of 301 men and women who entered a clinical trial. The results showed that the 2 factors had good reliability. Validity tests supported the interpretation of what the 2 factors measure, its implications for relationships to other variables, and the comparability of the 2- and 3-factor models. PMID- 11021161 TI - [Kidney transplant in the aged]. PMID- 11021162 TI - [Inflammatory cytokines]. PMID- 11021163 TI - [Antiretroviral therapy in HIV infection. Effects of the introduction of new drugs on health care expenditures, 1994-1999]. AB - The administration of antiretroviral compounds to our cohort of HIV-infected patients was assessed since 1994, on the ground of some epidemiological, clinical, and therapeutic variables. During the six-year study period, a significant increase of mean prescription rate of overall anti-HIV agents was observed, with a nearly 10-fold rise of mean prescribed daily doses per 1,000 patients-year. In particular, lamivudine and indinavir represented the most frequently administered drugs, among nucleoside analogues and protease inhibitors, respectively. A significant increase of the percentage of HIV infected patients undergoing combined antiretroviral therapy (79.3% in 1999), and the mean number of drugs prescribed per patient (3.02 in 1999), was concurrently detected. The progressive changes of antiretroviral therapy guidelines were responsible for a nearly 16-fold increase of expenditures directly related to antiretroviral drug administration in 1999 compared with 1994 (with over 41% of costs related to protease inhibitors). On the other hand, a substantial modification of HIV disease evolution occurred in our patient cohort in terms of absolute morbidity and mortality figures, as expressed by a drop of notified AIDS cases and AIDS-related deaths ranging from 2.5 to 5 times, during the considered period. PMID- 11021164 TI - ["The memory effect" in healthy heart and transplanted heart. "Cor se ipse alit, se ipse movet, se ipse reget"]. AB - The heart's memory, interpreted as information residue between a cardiac cycle and the subsequent one, is a very definite phenomenon (short- term memory) both in healthy subjects and in heart transplanted recipients. This "memory effect", we have observed by the autocorrelation and by the spectrum analysis of the values expressing the heartbeat acceleration variability (tachogram 24 h). The "memory effect" cannot be absolutely put down in heart transplanted recipients to interference or to the activity of the central and/or peripheral nervous system, because the heart transplanted is a denervated heart by definition. Moreover, an immediate regulating effect of hormonic factors in the short-term heartbeat regulation is not completely plausible. As already known, the heart transplanted responds to the emergency situation autonomously and autochthonously in proportion to the signals reaching it directly from the circle (vascular resistance, blood pressure, etc.). This implies the existence of functional memory that is likely to be inborn in the conduction system of the heart. Looking at the results of our research we can conclude: "cor se ipse alit, se ipse movet, se ipse reget" (it autonomously models itself to all the different modifications in the circle, it feeds itself as the organ propelling blood circulation and, if necessary, it carries on its kinetic activity autonomously). PMID- 11021165 TI - [Aspects of thrombophilia in Buerger's disease]. AB - Buerger's disease or thromboangiitis obliterans is a disorder of unknown etiology which affects young heavy smokers, mainly of male sex; it is not clear why this disease afflicts only a little number of heavy smokers. Studies about genetic predisposition, autoimmune phenomena and platelet function have been performed in order to search additional etiologic factors without finding significant alterations. Recently some cases of Buerger's disease in association with hypercoagulable states have been described. The authors reported ten subjects affected by Buerger's disease, who have undergone to a clotting process evaluation. We have found 3 cases with fasting hyperhomocysteinemia in association with low level of plasma folate, 3 cases with high value of lipoprotein(a) (in 1 subject in association with IgG anticardiolipin antibody positivity), 1 case with IgM anticardiolipin antibody positivity. These findings, along with literature data, induce to hypothesize that hypercoagulable states may favour the clinical expression of Buerger's disease. PMID- 11021166 TI - [Isolated acute renal failure secondary to sarcoidosis. Apropos of a case]. AB - A 59-years-old male patient was admitted to our Service because of acute renal failure with maintained diuresis (creatinine at admittance 6.2 mg/dl), preceded by malaise and weight loss. Clinical examination was normal and no investigation lead to a sure differential diagnosis of acute renal failure. The only abnormal laboratory investigations were: marked hypercalcaemia (12.7 mg/dl), slightly depressed parathormone (10 pg/ml) and anemia (Hb 11.2 g/dl). Also instrumental investigations performed were inexpressive. The abrupt appearance of an unilateral 7th cranial nerve paralysis lead to start a steroid therapy followed, in some days, by the normalization of calcium level and by a partial improvement of renal function. A renal biopsy was finally performed which permitted the diagnosis of interstitial granulomatous nephritis according to a sarcoidosis disease. Steroid therapy was continued allowing to a progressive, although not complete, recovery of renal function. The case peculiarity consists of isolated renal lesions with a severe expression of renal disease. The absence of classical disease criteria (pulmonary involvement firstly), in our case total body Gallium 67 scintigraphy was normal, should not exclude the diagnosis of sarcoidosis. The authors underline the importance of renal biopsy in detecting interstitial renal lesions potentially leading to uremia. PMID- 11021167 TI - [Guidelines for the treatment of dementias]. PMID- 11021168 TI - [Rehabilitation in Alzheimer's dementia]. AB - Alzheimer's disease is a neurodegenerative disease that causes a progressive decline of cognitive and behavioural functions. The simultaneous presence of these disorders requires a treatment not only for cognitive decline, but also for behavioural symptoms, depression and caregiver's stress. Research has made many efforts to develop a wide range of treatments, different from current pharmacological therapy, which is not resolutive, owing to the absence of an exact etiopathogenetic mechanism. Since new drugs have not been shown to be really effective in slowing cognitive impairment, various forms of rehabilitative interventions have been proposed in order to treat Alzheimer's disease. Their efficacy in the improvement of cognitive functions is still not completely clear. Surely, interesting results have been obtained from studies about Reality Orientation Therapy, Occupational Therapy and Memory Training. Music therapy might provide a new form of rehabilitative intervention, especially acting on the reducing of behavioural symptoms. These alternative forms of non pharmacological treatment may have a positive effect on caregiver. The heavy emotional burden of seeing a loved one becoming confused and isolated and of having to accept new responsibilities, may be reduced by rehabilitative supports, complementary to the pharmacological therapy. Caregiver stress could be reduced in two ways: by promoting the hope that something is being done for the patient and providing free time for himself. PMID- 11021169 TI - [Complications of lung transplantation: clinical aspects and radiological findings]. AB - In spite of improvements in the surgical technique of lung transplantation, early and late complications after surgery are not uncommon. The authors review the clinical aspects and radiological findings of these complications, including reperfusion edema, acute rejection, infections, airway complications, vascular complications, pleural complications, chronic rejection, omentopexy complications, transbronchial biopsy complications, post-transplantation lymphoproliferative disease and recurrence of the primitive disease. Early detection of these complications is important for early appropriate treatment, resulting in the decrease in morbidity and mortality of the transplanted patients. PMID- 11021170 TI - [Immortal music and mortal septicemias. Fatal sepsis of famous composers]. PMID- 11021171 TI - [A physician who can take care of the joints is the rheumatologist--a salutary lesson for the Japanese rheumatologists from the white paper issued by the Japanese Association of the Patients with Rheumatoid Arthritis]. PMID- 11021172 TI - [Clinical features of patients with osteoarthritic knees followed by development of rheumatoid arthritis]. AB - We investigated clinical features of patients with osteoarthritic knees followed by development of rheumatoid arthritis (RA) after several year's interval. The subjects were 16 knees of 8 patients with osteoarthritis (OA) including one man and 7 women. The mean age at development of OA knee was 62.8 years (range; 45 73). The mean age at later development of RA was 66.0 years (range; 52-79). The mean follow-up period was 96.4 months (range; 28-191). We evaluated clinical features using the 1987 revised Criteria of the American College of Rheumatology (ACR), laboratory dates including RF, CRP, ESR, the number of joints with RA, and femorotibial angle (FTA). The mean number of features of patients which was fulfilled with the ACR criteria was 3.3 +/- 1.6 at the onset of RA. Only four patients were seropositive through the total follow-up period. The serum level of RF, CRP, and ESR were reduced at the follow-up period. The mean number of the joints involved in RA was 11.0 +/- 5.1 (range; 4-22) and wrist and shoulder joints were involved more frequently than other joints except knees. High tibial osteotomy (HTO) was performed on 5 knees of 3 patients and the mean degree of FTA was 168.8 +/- 1.9 degrees just after surgery. However, 36 months after development of RA, joint destruction and valgus deformity occurred on 3 knees and the mean degree of FTA of 5 knees was ended up to 159.6 +/- 11.3 degrees. Our experiences suggested that RF, CRP, ESR and lesions of other joints should be carefully evaluated in the OA patients with seropositivity or knee hydrarthrosis and that histological analysis for synovium should be assessed by the biopsy at time of HTO or arthroscopic surgery to improve accuracy of diagnosis. PMID- 11021173 TI - [A case of rheumatoid arthritis associated with autoimmune hemolytic anemia due to weekly low-dose methotrexate therapy]. AB - A 57-year-old woman was found to have rheumatoid arthritis (RA) in 1996. Treatment with different immunoregulatory agents, including actarit and bucillamine, produced no improvement. Therefore, combined therapy with methotrexate (MTX) (5 mg/week oral) and low-dose prednisolone (PSL) (5 mg/day) was started in April 1997. Sulindac and famotidine were also administered. In August 1997, she was admitted to our hospital because of palpitations and shortness of breath due to severe anemia. Results of laboratory studies were hemoglobin, 2.9 g/dl; reticulocyte count, 225/1000; and haptoglobin, less than 10 mg/dl. The direct and indirect Coombs'tests were positive. A diagnosis of autoimmune hemolytic anemia (AIHA) was made on the basis of the laboratory findings. Treatment with high-dose PSL (50 mg/day) was started, and the anemia improved. The hemoglobin level increased to 6.0 g/dl within the 1st week and to 12.6 g/dl 6 weeks later. We believe that the most likely explanation for this anemia was the low-dose MTX because the anemia appeared soon after treatment was started. PMID- 11021174 TI - [A case of transient cortical blindness complicated by Henoch-Schonlein purpura with bronchiectasis]. AB - A 19-year-old woman had been treated for bronchiectasis since she was born. In October 1995, she was diagnosed as Henoch-Schonlein purpura (HSP) and HSP nephritis with the findings as follows; palpable petechial rash of legs, abdominal pain, arthralgias, and proteinuria. The administration of oral prednisolone was started, the clinical symptoms except for proteinuria was disappeared. However, nephrotic syndrome was continued despite the therapy of intravenous methylprednisolone pulse, various immunosuppressive drugs and warfarin. In February 1998, she was admitted to our hospital because of pneumonia. Several days later, her pneumonia improved on treatment with antibiotics, but she suddenly developed transient cortical blindness and acute renal failure. A provisional diagnosis of hypertensive encephalopathy and cerebral edema related to vasculitis was made, and she was treated with nifedipine, Glycelo, and high dose immunoglobulin. After the treatment, her vision and renal function had improved. She is a rare case associated with transient cortical blindness, bronchiectasis, and HSP. PMID- 11021175 TI - [A case of amyopathic dermatomyositis with rapidly progressive interstitial pneumonia]. AB - We report here a case of interstitional pneumonia (IP) associated with amyopathic dermatomyositis (DM). In August, 1998, a 53-year-old Japanese man was admitted to our hospital because of fever, polyarthritis and erythematous heliotrope eruption and Gottron's sign without any symptom of myositis. Serum CK level and EMG were normal. Jo-1 antibody was negative. Chest computed tomography (CT) revealed IP at both lung base areas. The patient was diagnosed as amyopathic DM with IP. When IP rapidly progressed, methylprednisolone pulse therapy and oral high dose prednisolone were not effective. High-dose intravenous cyclophosphamide seemed to be transiently effective, but oral cyclosporine A was not effective. The patient was died of respiratory failure in October 1998. During the course, we measured serum levels of LDH, sIL-2 R, s-ICAM-1 and KL-6, KL-6 could be a sensitive parameter of IP activity. PMID- 11021176 TI - [A case of intestinal Behcet's disease with abnormal ossification complicated by myelodysplastic syndrome, symptoms revealed after the perforation of ileum ulcer]. AB - A 39-year-old man, who had been treated with Etretinate for common wart since he was 29 years old, was admitted to Taga General Hospital complaining of gradually deteriorating lumbago and bilateral hip joints pain in September, 1996. His lower vertebrae and bilateral hip joints showed abnormal ossification on X-ray. The bone scintigraphy indicated the existence of sacroiliitis. His platelet counts were fluctuating between 8 x 10(4) and 9 x 10(4)/mm3. During the follow-up in our out-patient clinic, he was suddenly suffered from severe abdominal pain in August, 1997 and admitted to our hospital. An emergency operation revealed multiple ulcers of his ileum with several perforations. Histological findings of the specimen of the ileum showed simple ulcer. After the operation, he had oral and genital ulcers. He did not have any signs or symptoms of ocular involvement. He was diagnosed as intestinal Behcet's disease. Because he showed gradually pancytopenia for several months after the operation, bone marrow aspiration was performed and a diagnosis of refractory anemia, a type of myelodysplastic syndrome (MDS), with trisomy of chromosome 8 was made. Abnormal ossification of his vertebrae and hip joints were considered to be related to Behcet's disease because an coexistence with sacroiliitis. On the other hand, there is no denying the effects of orally Etretinate administration. Several cases have been reported the association of MDS with Behcet's disease. In this case, the existence of MDS or various symptoms in Behcet's disease became apparent after the perforation of ileum ulcer. This paper discusses possible etiology of the relation between Behcet's disease and MDS, or the characteristic clinical course in this case. PMID- 11021177 TI - [Etiology and high tibial osteotomy for osteoarthritic knee and regeneration of articular cartilage]. PMID- 11021178 TI - [Osteoporosis in rheumatoid arthritis]. PMID- 11021179 TI - [What is your diagnosis? Piezogenic nodules]. PMID- 11021180 TI - [Contraception from the viewpoint of women (2): Long-term pill users are especially pleased by the advantages of hormonal contraception. Comprehensive study of motives at first and refill prescription of the pill and evaluation after 3 months of hormonal contraception]. AB - The pill is the most famous contraceptive method beside the condom. It has a positive image both with women taking it for the first time as with women taking it again after an interruption: they appreciate its efficacy and its easy use. They consider a potential weight increase as a major disadvantage of the pill but do not fear serious health risks. Younger women are significantly more concerned by the lack of protection from AIDS. Young women start their "contraception carrier" with the condom and switch to the pill later. Middle-aged women had usually chosen the pill as first contraception method. The quality of life with the pill is considered as positive, especially its effects on skin and menses. PMID- 11021181 TI - [The value of conventional roentgen imaging and computerized tomography in diagnosis of frontobasal fractures]. AB - The classical x-ray diagnosis and the computer tomography (CT) examination depends mainly on clinical examination. The purpose of our study was to compare the value of conventional x-ray diagnosis and CT in detecting rhinobasal fractures. We examined 133 patients retrospectively who had a fracture. Altogether we considered with the classical x-ray diagnosis 111 and with the CT 250 regions of the frontobasis. In all, one third of fractures in the regions could not be detected preoperatively by CT and not the half by classical x-ray diagnosis. CT allows a direct detection of brain hernias and far better imaging of osseous lesions, also of fine structures, such as a better appraisal of paranasal sinus pathology. Classical radiology is, however, still indispensable for imaging certain fracture types and localization of a pneumocranium. Except in case of special demands, conventional x-ray diagnosis is important for the routine diagnostics of the facial skull as a primary method supplying an answer to the most important questions at a lower cost and in a shorter time. Altogether, CT allows an improvement of postoperative results, influencing indication, timing of operation, such as surgical approach. PMID- 11021182 TI - [Arterial hypertension and increased urinary catecholamines]. AB - We report the history of a 39-year-old-man who suffered from refractory arterial hypertension and who was referred to us for the diagnosis of phaeochromocytoma due to increased urinary free catecholamines. The work-up revealed the diagnosis of cocaine-induced arterial hypertension. The pathogenesis of cocaine-induced arterial hypertension as well as the differential diagnosis of increased urinary catecholamines will be discussed. PMID- 11021184 TI - [An unusual case of left-sided thoracic pain. Pleural tuberculosis]. PMID- 11021183 TI - [Leptospirosis: a frequently missed diagnosis? Epidemiology and diagnosis in Switzerland]. AB - Clinical leptospirosis is rarely seen in Switzerland due to the climatic conditions. Nevertheless people may still suffer from leptospirosis, even to the extent of requiring intensive medical treatment. Variable symptoms and problematic diagnostics can make the diagnosis of leptospirosis difficult. Through two cases with varied symptoms, the clinical and diagnostic features of the disease are presented. In addition, the risk factors, the epidemiology and the diagnostic possibilities are discussed. PMID- 11021185 TI - [Anal fissure--a new therapy concept]. AB - The anal fissure is one of the most frequent causes for anal pain. Conservative treatment usually consists of laxatives, local anesthetics and nitroglycerin cream. These therapies have a high recurrency rate. Surgical interventions, i.e. manual dilatation and sphincterotomy are fraught with the danger of fecal incontinence. The completely reversible effect of botulinum toxin injection opens new possibilities in the treatment of anal fissures. Its use is discussed as part of a 3-stage therapeutic regimen. PMID- 11021186 TI - [Sudden coma in a 20-year-old man]. AB - A 20-year-old male patient was brought to the emergency department by Emergency Medical Services after having been found unconscious. Upon arrival the patient was comatose with a GCS of 3, his vital signs were stable (with blood pressure 100/54 mmHg, heart rate 48 per minute, respiration rate 12 per minute and oxygen saturation 98% on room air). Both pupils were 3 mm, symmetric, and only minimally responsive. Approximately 2 hours after arrival the patient awoke and admitted having taken three ampoules of GHB (gamma hydroxybutyrate). GHB is a synthetic analog of gamma-amino butyric acid (GABA), a centrally inhibitory neurotransmitter. While GHB produces euphoria in low doses, small overdosing can result in severe poisoning with coma. The combination with other CNS depressants such as alcohol, opioids, and other narcotics is particularly dangerous. Physicians should be alerted to the clinical effects of GHB since abuse has become more widespread in Switzerland within the last months. In patients with unexplained coma the differential diagnosis of GHB-intoxication should be taken into consideration. PMID- 11021187 TI - [Diabetic nephropathy with nephrotic syndrome--apropos of 2 cases. Advantages of combined conversion enzyme inhibitor and non-dihydropyridine calcium antagonist treatment]. AB - The inhibitors of the angiotensin-converting-enzyme (ACEI) are considered as the best pharmacological class for the treatment of patients with diabetic nephropathy. Independently of lowering the arterial blood pressure they reduce the proteinuria and slow the evolution of the renal failure. Calcium-channels blockers not belonging to the dehydropyridine-type (verapamil-diltiazem) possess some of these features, too, contrarily to the rest of calcium-antagonists (nifedipine-like). Two clinical studies dealing with type-II diabetic-patients whose nephropathy was complicated by a nephrotic syndrome and a rapid progressive renal failure showed that the combination of verapamil with an ACEI, further dietetic measures (protein restriction, saltless diet), permitted a significant decrement of the proteinuria as well as a stabilisation of the kidney function. This effect could not be shown under treatment with only ACEI. Thus the proteinuria-inhibiting and kidney-protecting effects of the combination of theses two substances-groups should be known when the physician is confronted at this clinical situation that determines the prognostic of the kidney function in such a dramatic way and short laps of time. PMID- 11021188 TI - [Pain in the right epigastrium]]. PMID- 11021189 TI - [Galactorrhea and secondary amenorrhea in hyperprolactinemia]. PMID- 11021190 TI - [Psychological disorders and behavioral symptoms in patients with dementia]. PMID- 11021191 TI - [Neuropathological principles of behavioral disorders in dementia]. AB - Psychotic symptoms, apathy, agitation and aggressiveness are behavioral disorders that occur frequently in patients with Alzheimer's dementia. They cause serious problems for patients, relatives and care-givers. These behavioral disorders are associated with neuropathologic changes and alterations of brain metabolism in specific brain areas. Disturbances in mesotemporal and frontal brain areas seem to be related to psychotic symptoms. Apathy is associated with dysfunction of frontal cortical areas. Agitation and impulsivity appear to result from a hypofunction of the serotonin system in association with a relative hyperfunction of dopaminergic and noradrenergic systems. These dysfunctions are the result of direct neuropathologic changes, but also due to cholinergic deficits that seem to both contribute synergistically to and independently cause behavioral disturbances. PMID- 11021192 TI - [Psychological disorders and behavioral symptoms in dementia and differential diagnostic significance]. AB - Dementia is one of the most common diseases in the elderly. Rapid and comprehensive diagnosis and appropriate treatment are imperatives. Over the course of the disease, psychiatric disturbances and behavioral abnormalities will frequently occur with the most variable of manifestations and forms. The burden on caregivers is immense. Patients usually respond well to a combination of milieu therapy and pharmacological management. The nature and the manifestation of the disorder can provide important clues for differential diagnosis. PMID- 11021193 TI - [Behavioral disorders in patients with dementia--somatic causes]. AB - Behavioral disorders in dementia are often caused by somatic diseases. This suggests the importance of a focussed but careful physical examination, especially because it is often difficult to obtain correct anamnestic data. Its a fact that behavioral disturbances in dementia often improve by a successful treatment of somatic problems. This strategy may not only enhance the chance to find a nursing home where the patient can spend his further life but will also improve his quality of life and outcome. PMID- 11021194 TI - [Management of behavioral disorders in patients with dementia. Significance, diagnosis, non-medicamentous and medicamentous therapy]. AB - In the clinical course of dementia delusion, hyperactivity, depression, anxiety and aggression are very frequent behavioural disturbances. Caregivers are weighted down more by those disturbances than by the cognitive deficits of the patients. The hazy term of "agitation" may be seized diagnostically, differentiated by scales recording behaviour disturbances, as brain-organic causes of the agitation must be distinguished from both, reactive troubles and dementia specific disturbances. Regular medical assessment of the behavioural disturbances of demented patients is a starting point for rational environmental and medicinal interventions. The best prevention of disturbed behaviour is adequate treatment by not asking too much of the demented patients and an appropriate living ambience. Moreover there are newly developed atypical neuroleptics and the latest generation of antidepressants at disposal. A survey of the most prescribed drugs for the treatment of different symptoms is being explained. PMID- 11021195 TI - [Aging, love, sexuality--possibilities for psychotherapy interventions]. AB - Like high life expectancy also emancipation for questions of life quality and sexuality in old age is increasing. Estimations and possibilities for psychotherapy in old age would be painted out and discussed. PMID- 11021196 TI - [Developmental biotechnology: overview]. PMID- 11021197 TI - [Cloned mice from somatic cell nuclei]. PMID- 11021198 TI - [Animal cloning by somatic nuclear transfer]. PMID- 11021199 TI - [Microinsemination and nuclear transfer using male germ cells]. PMID- 11021200 TI - [Acquisition of developmental competence by nuclear transfer of immature oocytes]. PMID- 11021201 TI - [Stem cell technology: overview]. PMID- 11021202 TI - [Human pluripotent stem cell lines (ES and EG cell lines)]. PMID- 11021203 TI - [Molecular mechanism for cell-fate determination in ES cells]. PMID- 11021204 TI - [Purification and characterization of the hematopoietic stem cell]. PMID- 11021205 TI - [Neural stem cells]. PMID- 11021206 TI - [Generation of cardiomyocytes from mesenchymal stem cells]. PMID- 11021207 TI - [Liver stem cells]. PMID- 11021208 TI - [Cytokines for cell growth and tissue regeneration: overview]. PMID- 11021209 TI - [Functions of fibroblast growth factor and their receptors during morphogenesis]. PMID- 11021210 TI - [Therapeutic potential of HGF as a regenerative factor]. PMID- 11021211 TI - [The roles of TGF-beta superfamily in mouse early development]. PMID- 11021212 TI - [Epidermal growth factor]. PMID- 11021213 TI - [Neurotrophic factors]. PMID- 11021215 TI - [Biomedical technologies for tissue engineering: overview]. PMID- 11021214 TI - [Angiogenic inducers and inhibitors]. PMID- 11021216 TI - [Bioabsorbable polymer]. PMID- 11021217 TI - [Inorganic materials: porous hydroxyapatite for tissue engineering]. PMID- 11021219 TI - [Application of extracellular matrix biology to tissue engineering]. PMID- 11021218 TI - [Cell and cell sheet manipulation by utilizing biomaterials--development of cell sheet engineering]. PMID- 11021220 TI - [Immunoisolation]. PMID- 11021221 TI - [Drug delivery system]. PMID- 11021222 TI - [Cell culture engineering]. PMID- 11021223 TI - [Cryopreservation of biological material]. PMID- 11021224 TI - [Construction of organs and tissues and developmental engineering: overview]. PMID- 11021225 TI - [Role of dermo-epidermal interactions in the development of cutaneous appendages]. PMID- 11021226 TI - [Mechanisms of growth cone guidance]. PMID- 11021227 TI - [Kidney formation]. PMID- 11021228 TI - [Development and regeneration of skeletal muscle]. PMID- 11021229 TI - [Genetic plan for the formation of cardiovascular system]. PMID- 11021230 TI - [Organogenesis and regeneration of pancreas]. PMID- 11021231 TI - [Identification and enrichment of hepatic stem cells by flow cytometric cell sorting]. PMID- 11021232 TI - [Tissue engineering: overview]. PMID- 11021233 TI - [Hematopoietic stem cell transplantation]. PMID- 11021234 TI - [Regeneration of the central nervous system by cell transplantation]. PMID- 11021235 TI - [Peripheral nerve]. PMID- 11021236 TI - [Regenerative medicine of skin]. PMID- 11021237 TI - [Bone regeneration by cultured bone graft: tissue engineering using cultured marrow cells]. PMID- 11021238 TI - [Tissue engineered cartilage tissues]. PMID- 11021239 TI - [Hepatocyte transplantation for the treatment of hepatic failure]. PMID- 11021240 TI - [Implantation of bio-artificial endocrine pancreas]. PMID- 11021242 TI - [Present states and future perspectives of advanced medicine]. PMID- 11021241 TI - [Development of donor animals for xenotransplantation]. PMID- 11021243 TI - [Artificial organ]. PMID- 11021244 TI - [Human embryonic stem cell and ethical issues]. PMID- 11021245 TI - [Patents and bioethics: the problems posed by regenerative medicine]. PMID- 11021246 TI - [Current status of tissue engineering industry in Japan and U.S]. PMID- 11021247 TI - [Current trends of tissue regulation and guidelines]. PMID- 11021248 TI - [Accountability and public acceptance: role of massmedia]. PMID- 11021249 TI - Interventions in cardiology: what does and does not work. PMID- 11021250 TI - The comparative pathobiology of atherosclerosis and restenosis. AB - Percutaneous coronary interventions (PCIs) play an increasingly important role in the management of patients with coronary artery disease. However, these important procedures are complicated by restenosis in a sizeable number of patients. The pathobiology of atherosclerosis comprises a complex interaction among lipids, the endothelium, circulating and tissue inflammatory cells, platelets, and vascular smooth muscle cells. The superimposition of the mechanical and cellular consequences of PCIs on the abnormal substrate of atherosclerosis leads to a characteristic and distinct pathobiology that initiates and perpetuates restenosis. A clear understanding of the significant differences between atherosclerosis and restenosis will provide a rational basis for developing treatment plans that always address both problems. This article reviews and contrasts the pathobiology of atherosclerosis and restenosis and compares the mechanisms and time-course of these distinct entities. PMID- 11021251 TI - Rationale for a postintervention continuum of care: insights from intravascular ultrasound. AB - In 1987, Glagov published a provocative hypothesis that the principal initial response to plaque accumulation was enlargement of the vessel as a whole (remodeling), which preserved luminal diameter until the lesion occupied a substantial proportion of the area within the elastic external membrane (EEM). This model has been confirmed and extended by intravascular ultrasound (IVUS) studies. Remodeling renders most atheromas invisible to angiography. In patients with single-vessel disease, the mean percent of EEM area occupied by atheroma at a segment with an angiographically normal appearance is 39%. IVUS studies further show that the disease progresses in a diffuse manner and that stenotic lesions represent only a small proportion of the total disease burden. We further know that disease can begin as early as the teenage years. Data suggest that up to 25% of individuals < 25 years old may be affected. Atherosclerotic lesions can be effectively targeted with lipid-lowering therapy. Preliminary data suggest that as plaques stabilize and become smaller, a reverse modeling occurs: the EEM area decreases but the lumen size remains unchanged. This may explain why significant reductions in cardiovascular events in clinical trials were accompanied by only modest angiographic evidence of regression. This hypothesis will be tested prospectively in the Reversal of Atherosclerosis with Lipitor (REVERSAL) study. PMID- 11021252 TI - Lipid-lowering therapy after coronary revascularization. AB - Atherosclerosis is often asymptomatic, unrecognized, and undertreated. Lumen irregularities are important angiographic findings that should be addressed aggressively through risk factor modification, medical therapy, and coronary revascularization. Both angiographic and clinical benefits have been demonstrated with lipid reduction therapy in randomized clinical trials. Coronary revascularization is indicated for symptom relief and improvement in quality of life in patients with acute coronary syndromes at "intermediate" and "high" risk of subsequent death or myocardial infarction. In patients following percutaneous coronary intervention (PCI), future cardiac events may be related to lumen renarrowing or to progression of atherosclerotic disease at sites remote from the site of coronary revascularization. The time course of restenosis is relatively self-limiting, generally occurring within 6-12 months after the procedure. Clinical events occurring > 1 year after PCI generally relate to new lesions or progression of existing atherosclerotic disease. Patients with diabetes mellitus may be at higher risk for late coronary events than nondiabetic patients. In post coronary artery bypass surgery (CABG) patients, the majority of late events relate to degeneration of saphenous vein grafts. Lipid lowering therapy after coronary revascularization has been shown to prevent clinical events related to plaque instability and inhibit progression of saphenous vein graft disease. Thus, there are 2 goals in management of patients with symptomatic coronary artery disease: (1) to relieve the flow-limiting stenosis, and (2) to prevent future clinical events with aggressive lipid lowering and modification of other risk factors. Patients, specialists, and primary care physicians each need to take accountability for this risk-factor modification. PMID- 11021253 TI - Cholesterol lowering in atherosclerosis. AB - Lipid-lowering with statins reduces disease progression, prevents myocardial infarction and other hard end points, and prolongs survival. Data from large scale trials with these agents further show that lowering low-density lipoprotein (LDL) cholesterol in patients with coronary artery disease reduces the incidence of cardiovascular events and that the lower the LDL cholesterol achieved, the lower the event rate. Currently available evidence supports the National Cholesterol Education Program (NCEP) recommendations for reduction of LDL cholesterol levels to at least 100 mg/dL in patients with coronary artery disease. The Treating to New Targets study, which will evaluate the effects of LDL-cholesterol lowering to < or = 75 mg/dL with atorvastatin, may help clarify if additional benefit accrues with further reductions. However, up to 82% of patients with proven coronary disease are not even at the current NCEP lipid goal. Up to 55% need a > 30-mg/dL reduction in LDL cholesterol to reach that goal. These data suggest that many patients are not receiving a statin or are receiving an inadequate dose. Aggressive lipid lowering, although a desirable goal, does not yet appear to be standard practice. PMID- 11021255 TI - American National Standard--Personnel neutron dosimeters (neutron energies less than 20 MeV). American National Standards Institute, Inc. PMID- 11021254 TI - Collectins and collectin receptors in innate immunity. AB - This thesis is based on nine papers and a review on the collectins and collectin receptors in innate immunity. The collectins are a family of proteins in which the individual chains consist of a C-type lectin domain attached to a collagen domain via an alpha-coiled neck region. The chains are organized into a triple collagen helix and oligomerized through N-terminally located cysteines. The collectins have a dual function: one is to bind specifically to carbohydrate structures on the surface of a pathogen; the other is subsequently to recruit other cells and molecules to destroy the pathogen. The C-type lectin domains contain 110-130 amino-acid residues arranged in a conserved sequence pattern which allows the domain to fold into a well-defined tertiary structure. Five collectins have been described. Lung surfactant proteins A and D (SP-A and SP-D) are mainly found in the surfactant coating the luminal surface of the pulmonary epithelial cells, but are also produced by cells lining the gastrointestinal tract. Mannan-binding lectin (MBL), conglutinin and collectin-43 (CL-43) are serum proteins produced by the liver. Conglutinin and CL-43 have so far only been found in Bovidae. The collectins are involved in innate, nonadaptive immune defense. They bind to microbial surface carbohydrates, inducing aggregation and thereby impeding infectivity or mediating phagocytosis through specific receptors on the phagocytes. After binding microbial carbohydrate, MBL can activate the complement system through a newly discovered pathway which makes use of two serine proteases (MASP-1 and MASP-2) to activate the complement factors C4 and C2. In man, low serum MBL concentrations resulting from mutations in the collagen region are associated with a common opsonic defect. CL-43 was identified as a new collectin by its calcium-dependent binding to mannan and by its M(r) of 43 kDa in the reduced state on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The lectin was isolated by affinity chromatography on mannan Sepharose, absorption with rabbit anti-bovine Ig coupled to Sepharose-4B and ion exchange chromatography. CL-43 shows an apparent molecular mass of 120 kDa in the unreduced state on SDS-PAGE and elutes with an apparent molecular mass of 750 kDa on gel chromatography under nondissociating conditions. Amino-acid analysis and susceptibility to collagenase digestion indicated that CL-43 was a collectin. Electron microscopy of purified CL-43 revealed only rod-like monomer subunits 37.4 nm long. Two-dimensional gel electrophoresis showed that CL-43 has two isoforms of pI 4.9 and 5.3 respectively, corresponding to the native form of CL 43 and a truncated form which lacks the first 9 amino-acid residues. The N terminal amino-acid sequence of CL-43 was used to design primers for PCR with a bovine liver cDNA as template. The cDNA of CL-43 was cloned and the open reading frame was found to encode a protein of 301 amino-acid residues, including an N terminal region of 28 residues, a collagen region of 114 residues and a neck-CRD region of 159 residues. The amino-acid sequence of CL-43 shows 74% identity with bovine conglutinin and 70% identity with bovine SP-D, but the collagen region is considerably shorter than those of conglutinin and SP-D. Northern blot analysis showed that CL-43 was only synthesized in bovine liver, no signal being detected in a variety of other bovine tissues, including lung. No cross-hybridizing signals were detected in mRNA from ovine, human, rat or mouse liver. Since CL-43 and conglutinin have only been detected in members of the Bovidae, it is probable that an ancestral gene of these two proteins was first derived from a SP-D-like gene and that this ancestral gene underwent duplication during evolution. The carbohydrate binding profile of CL-43 was analyzed by an inhibition assay with biotinylated CL-43, using solid-phase mannan as the ligand. (ABSTRACT TRUNCATED) PMID- 11021256 TI - How safe is our drinking water? PMID- 11021257 TI - Improving detection of venous thromboembolism. New technology holds promise for early, precise diagnosis. AB - In recent years, a number of modalities have been evaluated for the diagnosis of venous thromboembolism. The role of these modalities is still evolving. While ventilation-perfusion scanning is important in the diagnosis of venous thromboembolism, spiral CT scanning, MRI, and D-dimer assays are now being used more often, either exclusively or in combination with ventilation-perfusion scanning. A number of diagnostic algorithms using these modalities are currently being evaluated. Regardless of which diagnostic approach is used, the clinician must be aware of some key limitations. Spiral CT scanning has gained popularity because it is noninvasive and can rapidly identify other cardiopulmonary diseases that mimic pulmonary embolism. Its use has been limited because of its inability to detect subsegmental pulmonary emboli. D-dimer assays offer promise as rapid, inexpensive screening tools. However, the wide variability in assay performance has limited its usefulness. We recommend that if D-dimer assays are to be used in a diagnostic algorithm, the clinician be aware of the details of the assay. At present, lack of data precludes use of MRI as a primary diagnostic tool for detection of venous thromboembolism. PMID- 11021258 TI - Anticoagulation for venous thromboembolism. What are the current options? AB - Venous thromboembolism remains an important cause of considerable morbidity and mortality. Low-molecular-weight heparin appears to be a safe and effective alternative to unfractionated heparin for inpatient treatment. In addition, we recommend considering its use in outpatient treatment in selected patients. When warfarin therapy is initiated, the starting dose should approximate the suspected maintenance dose. The optimal length of anticoagulation after an initial episode of venous thromboembolism is 6 months unless a persistent risk factor is identified. Thrombolytic therapy for hemodynamically stable patients remains controversial, primarily because of the potential for devastating complications. PMID- 11021259 TI - Evaluation of the hypercoagulable state. Whom to screen, how to test and treat. AB - Venous thromboembolism is a common disease that causes significant morbidity and mortality. In recent years, the ability to diagnose inherited genetic defects and common acquired conditions predisposing to thrombosis has greatly increased. Venous thromboembolism is now understood to be a complex interaction of genetic and environmental factors leading to thrombosis. Integrating the various factors to individually assess thrombotic risk still poses a challenging clinical problem that will likely become easier as more data accumulate. As the ability to accurately assess risk increases, the data can then be translated into tailored treatment regimens. Until then, only general guidelines regarding evaluation and management are available. In the future, it is likely that other prothrombotic conditions will be elucidated, adding to the pool of data. PMID- 11021260 TI - Thromboembolism during pregnancy. Risks, challenges, and recommendations. AB - Pregnancy is an important risk factor for venous thrombosis, and venous thromboembolism is a leading cause of preventable death in pregnancy. Diagnosis of venous thromboembolism is complicated in that the symptoms of dyspnea and lower extremity edema are relatively common complaints of pregnant patients. Physicians should maintain an appropriately high index of suspicion and request diagnostic imaging in a timely manner. Diagnosis of deep venous thrombosis with Doppler ultrasonography of the lower extremity poses no health risk to the fetus, but other radiographic studies pose a low radiation risk to the fetus. Because anticoagulant therapy poses a greater health risk to mother and fetus than does the radiation required for the diagnosis of pulmonary embolism, clinicians should aggressively pursue objective evidence of venous thromboembolism. Once the diagnosis is made, anticoagulation with intravenous unfractionated heparin or subcutaneous low-molecular-weight heparin should be used prepartum followed by warfarin therapy after delivery. PMID- 11021261 TI - Common dermatoses of the male genitalia. Recognition of differences in genital rashes and lesions is essential and attainable. AB - Dermatoses of the male genitalia can be confusing to identify and difficult to diagnose and treat. Rashes and lesions that occur on other areas of the body can be hard to recognize when they appear on the genitalia. In this article, Dr Goldman reviews the common dermatoses, presents defining characteristics, and suggests treatment options. PMID- 11021263 TI - What's causing these scalp nodules? Cutaneous metastasis. PMID- 11021262 TI - Sports supplements. Can dietary additives boost athletic performance and potential? AB - Nutritional sports supplements, many of which are endorsed by professional athletes, are increasing in popularity among casual and adolescent sports enthusiasts, bodybuilders, and weight lifters. Because many people consider nutritional additives to be "natural" and therefore "safe," patients may not consider the possible effects of those substances when taken in high doses or in combination with medications. Drs Rubinstein and Federman present an interesting overview of several sports supplements and examine the consequences and caveats of their use, as well as the reasons for their popularity. PMID- 11021264 TI - Is it multiple sclerosis? PMID- 11021265 TI - Management of questionable hypothyroidism? PMID- 11021267 TI - IFCC-WorldLab Satellite meeting n. 9: Aging and Clinical Laboratory. Alghero, Italy, 11-13 June 1999. Abstracts. PMID- 11021268 TI - IFCC-WorldLab. Firenze, Italy, 6-11 June 1999. Abstracts. PMID- 11021266 TI - 32nd Annual meeting of the Society for the Study of Reproduction. July 31-August 3, 1999. Pullman, Washington, USA. Abstracts. PMID- 11021269 TI - Plant topics. Society for Experimental Biology annual meeting. 27-31 March 2000. Abstracts. PMID- 11021270 TI - [Hormone therapy in heart failure: growth hormone and insulin-like growth factor I]. AB - Heart disease is the major cause of mortality in the developed world. Despite recent advances in the therapy of heart failure due to ACE inhibitors and beta blockers, the prognosis of this syndrome is still poor. In the past few years, the effects of growth hormone (GH) and insulin-like growth factor-I (IGF-I) on heart morphology and function were extensively studied. Some studies dealing with experimental heart failure of animals and one controversial study dealing with human heart failure suggest positive hemodynamic effects of GH and/or IGF-I treatment. This review summarizes the physiological effects of GH/IGF-I on the myocardium, their signal transduction mechanisms, and the data currently available on the therapeutic use of these agents. PMID- 11021271 TI - [Hypocalcemia-induced heart failure as the initial symptom of hypoparathyroidism]. AB - Hypocalcemia is a relatively uncommon reversible cause of congestive heart failure. There are a few reports of hypocalcemic children who developed congestive heart failure associated with hypoparathyroidism. In all these patients, however, cardiac failure did not occur before the age of nine years. In addition, other striking noncardial manifestations of hypoparathyroidism, e.g., convulsive seizures, had been present prior to cardiac symptoms. We report on a 3.7 year old girl with mitral insufficiency and severe cardiac failure due to hypocalcemia secondary to familial hypoparathyroidism. The infant's mother was suffering from idiopathic hypoparathyroidism, but her own history lacked any evidence for parathyroid hormone deficiency. On admission, she presented with fatigue, dyspnea, and pedal edema. Liver edge was palpable 4 cm below the right costal margin, and a 3/6 systolic murmur was heard. A chest x-ray showed cardiac enlargement; electrocardiogram demonstrated a prolonged QTc interval of 0.46 s. The echocardiography revealed a cleft in the mitral valve with mitral insufficiency and markedly reduced contractility of the left ventricle. Laboratory studies demonstrated a low total serum calcium level of 1.3 mmol/l; serum magnesium level was slightly decreased (0.5 mmol/l), and parathyroid hormone level was not detectable. Partial monosomy of chromosome 22 was excluded. Ophthalmological examination, audiometry, and renal ultrasonogram were normal. Oral calcium supplementation and anticongestive therapy with metildigoxin, furosemid, and captopril was initiated but no improvement of the heart failure occurred. However, normalization of serum calcium level by calcium infusions caused prompt clearing of the clinical symptoms, complete normalization of liver size, reduction of cardiac enlargement (thoracic ratio decreased from 0.68 to 0.57), and marked improvement in contractility (left ventricular shortening fraction increased from 21% to 34%). The QTc interval decreased to 0.39 s. The successful treatment following normalization of serum calcium level proved the superiority of hypocalcemia over mitral valve insufficiency in the etiology of the cardiac failure. To our knowledge, this is the first report of congestive heart failure due to hypocalcemia as the first manifestation of hypoparathyroidism in childhood. Hypocalcemia should be kept in mind in any congestive heart failure in children with or without underlying cardiac malformation. PMID- 11021272 TI - [1998 prognosis of left ventricular dysfunction in a maximum care non-university hospital. Analysis of the Ludwigshafen LVD Registry]. AB - BACKGROUND: Prognostic data on heart failure rely on epidemiological studies or large clinical trials. These data are not to transfer easily on everyday practice. OBJECTIVE: To assess the prognosis of left ventricular dysfunction under the therapeutic conditions of the 1990s. SETTING: Nonuniversity tertiary care hospital. PATIENTS AND METHODS: Since January 1995 consecutive and complete registry of all inpatients presenting with a left ventricular dysfunction (EF < 45%). No exclusion criteria. RESULTS: n = 512, mean age 64 years, 77% male, mean NYHA class 2.5, left ventricular EF 31%. ETIOLOGY: 58% coronary heart disease, 28% dilated cardiomyopathy. Medical treatment consisting of 91% ACE inhibitors, ACE inhibitor dosage 52% of the recommended dose, 42% beta-blockers, 70% diuretics, and 63% digitalis. Follow-up completeness 95.5%. One-year mortality 64/489 patients (13.9%). Determinants of mortality: NYHA III/IV, EF < 30%, malignant disease, age > 75 years. CONCLUSION: In spite of the aforementioned medical treatment and a proportion of 14.8% implanted defibrillators in 1998, the one-year mortality of ventricular dysfunction is as high as 13.9%. In everyday practice the prognostic effect of ACE inhibition and beta-blockade is not completely utilized. PMID- 11021273 TI - [Using the internal thoracic artery of patients with reduced left ventricular function]. AB - Todate internal mammary artery (IMA) is routinely used in coronary artery bypass grafting even in elder patients. However in patients with poor left ventricular function use of the IMA is discussed controversely in Germany. Main arguments against IMA are an increased operation time, initially lower blood flow, higher rates of reoperation for bleeding and more perioperative complications. In this study we investigated use of the IMA in patients with poor left ventricular function (LVEF < 40%) compared to exclusively veingraft bypass surgery. 137 patients (105 m/32 f) suffering from coronary artery disease with reduced LVEF (12-40%) were randomized in the study. 67 patients received exclusively vein grafts (group I), 70 patients routinely obtained an IMA graft. Criteria used for evaluation of IMA graft were operation time, postoperative bleeding, need for catecholamines, requirement of intensive care, perioperative myocardial infarction and mortality. The number of distal anastomoses in each group was 3.1 (2-5). The operation time varied in compliance with the number of distal anastomoses, but there were no significant differences between both groups. Postoperative bleeding until the second postoperative day was 905 ml in group II versus 569 ml in group I; the difference was significant (p < 0.05). The need of catecholamines after operation and hemodynamic parameters were comparable in both groups, there were no significant differences. Intensive care was required for a mean of 1.6 days in both groups, postoperative ventilation was 5.8 hours in group I versus 7.9 hours in group II, differences not significant. Ischemia or myocardial infarction could be demonstrated in 2 patients of group I (3%) versus 4 patients of group II (5.7%). The differences between the groups were not significant. Cardiac low output syndromes without sights of myocardial infarction were apparent in 9 patients of group I (13.5%) versus 2 patients of group II (2.9%), this difference being significant (p < 0.05). Mortality after operation in both groups was higher than in patients with normal ventricular function, however the differences between the evaluated groups were not significant (5.9% in group I versus 4.3% in group II). Summarizing the above it can be concluded that patients with poor left ventricular function are at a higher risk when subjected to bypass operation; the use of IMA did not show any disadvantages in comparison to exclusively veingraft surgery, except of a higher perioperative bleeding risk. Due to better long term results IMA should be used routinely also in bypass-patients with poor left ventricular function. PMID- 11021274 TI - [Large pseudoaneurysm of the left ventricle after posterolateral wall infarct]. AB - Pseudoaneurysms of the left ventricle are rare complications after acute myocardial infarction. We report on a 69 year old patient with a large false aneurysm located in the posterolateral ventricular wall. It became clinically apparent during an episode of severe left heart failure. Echocardiography and left ventriculography allowed an accurate determination of localization and dimension of the pseudoaneurysm; coronary angiography revealed a proximal occlusion of the left circumflex coronary artery. Because of the high risk of rupture, a rapid surgical repair of the false aneurysm was indicated. PMID- 11021275 TI - [Need for ambulatory cardiology after-care rehabilitation in the metropolitan area of a large city. Results of the Cologne model of ambulatory cardiologic rehabilitation--phase II]. AB - Within the Cologne Model (CM) of outdoor cardiac rehabilitation (OCR), phase II investigations about the demand for this form of cardiac rehabilitation (CR) after acute cardiac diseases were carried out in three general hospitals, the cardiological and cardiosurgical university hospitals of Cologne. The subsequent questions were investigated: total number of coronary or cardiac operated patients, number of patients with indication for CR, and number of patients corresponding to the restricted indications of CM (age below 65, low risk patient, no cardiac operation). For these groups the acceptance or refusal of CR was checked. Subsequently a sample of patients corresponding to the criteria of CM or of operated patients were confronted with the additional offer of an OCR. The motivation for the acceptance of rehabilitation in specialized hospitals (ICR), OCR or refusal of each kind of CR was inquired. The acceptance of CR in the different groups varied widely. Whereas operated patients in Cologne accept CR in nearly 100% of cases, this is the case in patients after acute myocardial infarction (AMI) in only 50% and in patients after PTCA without AMI in only 5-6%. The analysis of predictors for acceptance brings about that younger patients prefer CR, and if they do, OCR. Patients with the more serious form of disease prefer ICR. Women accept CR more rarely than men, and if they do, they prefer the hospital form. However, this is less gender specific but consequence of the generally more serious form and later onset of CAD in females. Higher educational as well as occupational status favors acceptance of CR and specially OCR. The suspicions that unmarried people prefer OCR and foreigners ICR could not be generally confirmed. Crucial reasons for the form of CR which is accepted or refused are individual ones. ICR is favored by the wish for more safety and better recuperation. For OCR, the comfortable conditions at home with high social support and/or antipathy against hospitals after long clinical treatment are named. Analysis of demand for OCR demonstrates that between 40% in low risk patients (corresponding to CM criteria) and 20% in more serious cases (operated patients) prefer the outhospital form. From these data an estimation of demand for OCR in areas with high population was carried out. PMID- 11021276 TI - [Sudden cardiovascular death during sexual intercourse--results of a legal medicine autopsy study]. AB - This epidemiologic, retrospective follow-up mortality study is based on the data gathered in the department of forensic medicine at the University Hospital of Johann Wolfgang Goethe University in Frankfurt/Main. Over a period of 21 years (1972-1992), roughly 21,000 forensic autopsies revealed 39 cases (0.19%) of natural deaths occurring during sexual activity. Except for two women, all of these cases were men of an average age of 61.3 years. The most frequent cause of death was myocardial infarction (n = 21/53.8%), three of these cases involving pericardial tamponade accompanied by myomalacia. The medical history of twelve of the deceased showed a previous myocardial infarction. Most of the deaths took place during or after sexual intercourse or manual stimulation. In most cases sudden death occurred during the sexual act with a prostitute. The annual incidence of sudden cardiovascular deaths during sexual activity is estimated to be 0.2/100,000 men. The risk for women is lower by a factor of 12. PMID- 11021278 TI - [In Process Citation] PMID- 11021277 TI - 19th World Congress of Dermatology. 15-20 June 1997, Sydney, Australia. Abstracts. PMID- 11021279 TI - [Technics for exploring the optic nerve]. AB - This issue propose an inventory of the methods of investigation which help for the diagnosis of the optic neuropathies. These investigation methods are shortly presented, and the situation in which there are usefull are presented. Some of these investigation are expensive and are not available in any ophthalmological hospitals. Despite of the great number of investigations, the problem of the etiopathogenesis of the optic neuropathies is not completely elucidated. PMID- 11021280 TI - [Modern pathogenetic-therapeutic approaches in glaucoma]. AB - The review presents briefly the new orientations in pathogenically research of glaucoma, the actual theory of apoptosis in generating glaucomatous neuropathy. The substances and transmitters that interfere in the process of apoptosis are also discussed. It is also discussed the new neuroprotective medication. The conclusion is that this new therapy will be an adjunctive one to the classical antiglaucomatous therapy. PMID- 11021281 TI - Seven steps to assure the best possible treatment of persons with glaucoma or suspected of having glaucoma. AB - Glaucoma causes more blindness and disability than any other eye disease except for cataract. Cataract surgery is highly successful, even intracapsular cataract surgery. The major problem, then, with regard to relieving or preventing disability related to cataract is one of access to care. Though access to care is a major problem with glaucoma, it is by no means the only problem. Many patients with glaucoma get worse, even when they are being treated. Additionally, it is not only glaucoma itself that causes disability. In the recent past, far more damage was caused by treatment for glaucoma than by glaucoma itself. To repeat, summarized slightly differently, glaucoma and its treatment damage health more than any other eye disease except cataract, and the medical profession is responsible for a major part of that problem. How can it be that glaucoma and its treatment still cause so much disability? In the first place, care is unavailable in many areas, and, second, care is not utilized even where available, and third, care is often inappropriate even where available. The first two issues are of great importance but are not the subject of this discussion. It is my belief that the major explanation for why care is often poor is because the goals of care are often wrong. Specifically, the goals of care are to lower intraocular pressure, or to prevent visual field loss. These may be appropriate intermediary steps on the way to achieving the appropriate goal, but they are not the goal itself. PMID- 11021282 TI - [Juvenile glaucoma]. AB - Juvenile glaucoma is an uncommon form of chronic open angle glaucoma that appears between 3 and 35 years of age. We reports in this study four cases; one of them had unilateral visual acuity reduced. The intraocular pressure is above to 30 mm Hg, and a myopia was associated to two cases. The insidious development of this pathology and the difficulty of its diagnosis among young people, often result in severe clinical manifestations, with high visual field defects, and important optic disc cuppings. PMID- 11021283 TI - [The vitreoretinal complications of cataract surgery]. AB - The paper reports a retrospective study performed between January 1996 and January 1999 which comprised 423 patients who underwent eye cataract surgery. Vitreoretinal complications (retinal detachment, cystoid macular edema, choroidal detachment, endophthalmitis, luxation of the lens into the vitreous body) were related to age, sex, other general and local diseases and surgical complications, like disruption of the posterior capsule. The study presents also vitreoretinal complications after Nd:YAG laser posterior capsulotomy. The outcomes of the study show that 18 cases (4.25%) had retinal detachment, 11 (2.60%) had cystoid macular edema, 2 cases had endophthalmitis, while choroidal detachment and luxation of the lens into the vitreous body were reported each in 1 case. Retinal detachment following Neodymium:YAG laser capsulotomy (performed in 109 cases) was found in 2 cases. All of those vitreoretinal complications were reported with a higher incidence in patients with vascular diseases (diabetes mellitus, arterial hypertension, angiosclerosis) and also in patients who had axial lengths of 25 mm or greater. PMID- 11021284 TI - [Potentiated loco-regional anesthesia in ophthalmological surgery]. AB - A clinical study on 46 patients, who were operated on for ophthalmologic diseases under local anaesthesia, is presented. Midazolam, diazepam and/or pethidine, given to the patients 5-10 minutes before the operation improved the classical local anaesthesia; they ensured hypnosis, analgesia, anterograde amnesia and vegetative protection, thus offering a high surgical comfort and diminishing considerably the rate of the well known incidences and complications of the local anaesthesia. PMID- 11021285 TI - [The clinico-statistical aspects in allergic conjunctivitis]. AB - Ocular allergy represents the hypersensitivity reaction of the ocular surfaces in contact with the external environment. Allergic conjunctivitis are one of the frequent disorders of the clinical practice. In this paper we present statistically the cases attended in our service for one year, a few pathogenetical factors; the positive diagnose and the therapeutic strategy. PMID- 11021286 TI - [The efficacy of acyclovir treatment in the therapy of herpetic keratitis]. AB - There were investigated 51 patients, 19 female and 32 male, with ages between 12 and 80 years, hospitalized in the Dept. Ophthalm. of the City Hospital of Arad, during 1995-1997. From these, 21 had the clinical diagnosis of superficial keratitis, typical form of dendritic ulcer, 7 had non-typical forms, and 4 had stromal keratitis; 19 patients recurred. The diagnosis of herpetic keratitis was established correlating the clinical aspect with the cytologic examination, made on conjunctivo-corneane smears. A number of 32 patients received a specific treatment with Acyclovir, in an ointment form in local applications and on general route orally; there were also associated mydriatics and epitelizants of the cornea. The evolution of the disease was evaluated by comparing results obtained in different groups of patients. PMID- 11021287 TI - [Prostaglandin esters--new directions in the treatment of glaucoma]. AB - The therapeutically approach of glaucoma still represent a problem of the ophthalmology. Latanoprost, a new prostaglandin derivative, fall the intraocular pressure as good in the night as during the day by its special way of action. Since January 1998, in the Eye Clinic of the Central Military Hospital, we studying the ocular hypotensive effects of the latanoprost. Our results seems to confirm the efficacy of this prostaglandin analogue. PMID- 11021288 TI - [Congenital glaucoma and trabeculectomy]. AB - The authors presents 27 cases (48 eyes) with congenital glaucoma operated with trabeculectomy. We used 3 different procedures of trabeculectomy: suture of the scleral flap in two points; by placing a silicon membrane under the scleral flap; tightly close of the scleral flap with five sutures. We concluded that the last of the procedures gave us the best results, especially when was used in the incipient stage of the disease. From this study results that the mechanism of action of trabeculectomy is more an opening of a physiological way of drainage then a filtration to the subconjunctival space. PMID- 11021289 TI - [The therapeutic aspects in intermittent divergent strabismus]. AB - PURPOSE: Is to evaluate the possibilities of treatment in intermittent divergent squint at children and young adults. MATERIALS AND METHOD: We studied an group of 66 patients treated in the University Clinic of Ophthalmology from Cluj-Napoca. RESULTS: The most frequent type of intermiTtent divergent squint (IDS) is the type of excess of divergency (25.76%). In 46.9% the onset was between 2-6 years. In 69.7% visual acuities were equal and stereopsis was present in 74.24%. Hyperopia +/- astigmatism was present in 69.7%. The treatment of intermittent exodeviation was conservative in 51.5% and surgical for first intention in 39.3%. CONCLUSIONS: 1. The most frequent type of intermittent exodeviation is by excess of divergency. 2. The treatment of exodeviation is an optical, orthoptic and surgical treatment. 3. The surgical technique depends on the type of intermittent exodeviation. PMID- 11021290 TI - [Cytomegalovirus retinitis and optic neuropathy in a case of an infectious HIV syndrome]. AB - It is presented a case report of HIV syndrome complicated with cytomegalovirus retinitis and anterior ischemic optic neuropathy. It is reported a 36-years-old with type-B hemophilia and HIV-positive since few years. Despite treatment not recovered of ocular findings is obtained. Some literature data about ocular complications in HIV syndrome, particular features of CMV retinitis and anterior ischemic optic neuropathy etiopathogenesis are discussed. PMID- 11021291 TI - [A clinical case of functional recovery after an extensive, perforating corneoscleral wound with partial evisceration of the membranes and crystalline lens]. AB - The article presents a clinical case of functional recuperation after perforated corneo-scleral wound with partial evisceration of the eye and lens. The authors describe the possibilities of evolution and the surgical modalities for correction of aphakia. PMID- 11021292 TI - [The chorioretinal location of Toxoplasma infection]. AB - Considered rare and occurring only in immunocompromised patients or patients with severe infection, Toxoplasma gondii chorioretinitis--as a unique presenting sign of toxoplasmosis appears to occur with increased frequency over the last few years in immunocompetent patients. The typical clinical picture (central chorioretinal lesion with recurrent vasculitic reactions) and positive IgG Toxoplasma gondii serology confirmed our diagnosis. Negative Toxoplasma IgG serology in the mothers of 2 patients excluded the possibility of congenital toxoplasmosis. Clinicians should be aware of the possibility of increased frequency of ocular toxoplasmosis in immunocompetent patients. PMID- 11021293 TI - European Society for Comparative Physiology and Biochemistry 21st Congress. Liege, Belgium, July 24-28, 2000. Abstracts. PMID- 11021294 TI - The American Academy for Cerebral Palsy and Developmental Medicine (AACPDM) 2000 meeting. Abstracts. PMID- 11021295 TI - Congress of the European Association of Nuclear Medicine. 2-6 September 2000, Paris, France. Abstracts. PMID- 11021296 TI - Cleft 2000 meeting of the International Cleft Lip Palate Foundation. Abstracts. PMID- 11021297 TI - 25 year cumulative index 1973-1997. PMID- 11021298 TI - Society for Leukocyte Biology 34th annual meeting: Interactions of innate and acquired immunity. Cambridge, Massachusetts, USA. October 5-8, 2000. Abstracts. PMID- 11021299 TI - Society for Leukocyte Biology membership directory. PMID- 11021300 TI - 9th International Symposium on Celiac Disease. Hunt Valley, Maryland, USA. August 10-13, 2000. Abstracts. PMID- 11021301 TI - [Proceedings of the V Congress of the International Association of Morphologists. Abstracts]. PMID- 11021302 TI - Docetaxel in the management of breast and non-small-cell lung cancers. ASCO 2000, 22nd annual San Antonio Breast Cancer Symposium, and ECCO 10. Abstracts. PMID- 11021303 TI - 1st Congress of the International Pediatric Transplantation Association. Venice, August 23-25, 2000. Abstracts. PMID- 11021304 TI - Ca-activated K channels: the ins and outs of calmodulin. PMID- 11021305 TI - Hemoglobin affinity for oxygen in three subspecies of toads (Bufo sp.) living at different altitudes. AB - Blood oxygen affinity and red blood cell properties were measured in three subspecies of genus Bufo: Bufo spinulosus limensis, collected at sea level and at an average day temperature of 20 degrees C; Bufo spinulosus trifolium, from 3100 m, average day temperature of 15 degrees C; and Bufo spinulosus flavolineatus, from 4100 m, average day temperature of 10 degrees C. Electrophoresis of the hemoglobin showed the same component in each of the three subspecies. At 20 degrees C the blood oxygen affinities (P50) showed small differences between Bufo spinulosus limensis and Bufo spinulosus trifolium, whereas the value for Bufo spinulosus flavolineatus was markedly lower. At 10 degrees C, the ambient temperature of Bufo spinulosus flavolineatus, the P50 was extremely low compared with the other two subspecies at their corresponding ambient temperatures. PMID- 11021306 TI - Characterization and mapping of an informational suppressor in Aspergillus nidulans. AB - The present work was undertaken to characterize a suppressor gene present in a mutant strain of A. nidulans obtained with NTG (N-Methyl-N'-Nitro-N Nitrosoguanidine). Analyses of this mutant have shown that this suppressor, designated suO1, induces phenotypic co-reversion of several auxotrophic mutations and makes the strain sensitive to aminoglycoside antibiotics and lower temperatures. suO1 has shown to be on linkage group VIII. The vegetative growth of the mutant strain is very unstable because the suppressor gene induces the production of prototrophic mitotic sectors. The strains bearing the suO1 gene produce cleistothecia containing a reduced number of viable ascospores during the sexual cycle. The segregation of the genetic markers has also been observed in the mutant strain self crossed. From the above results it may be concluded that suO1 is an informational suppressor. PMID- 11021307 TI - Zinc fingers: DNA binding and protein-protein interactions. AB - The zinc finger domain is a very ubiquitous structural element whose hallmark is the coordination of a zinc atom by several amino acid residues (cysteines and histidines, and occasionally aspartate and glutamate). These structural elements are associated with protein-nucleic acid recognition as well as protein-protein interactions. The purpose of this review is to examine recent data on the DNA and protein binding properties of a few zinc fingers whose three dimensional structure is known. PMID- 11021308 TI - Ontogenetic changes in the fractal geometry of the bronchial tree in Rattus norvegicus. AB - Respiration and metabolism change dramatically over the course of the development of vertebrates. In mammals these changes may be ascribed to organogenesis and differentiation of structures involved in gas exchange and transport and the increase in size. Since young as well as mature individuals must be well-designed if the species is to survive, the physiological changes during the development should be matched with geometrical or structural adjustments of the respiratory system. The aim of this study was to evaluate changes in the fractal geometry of the bronchial tree during the postnatal development of the rat. The average fractal dimension of the bronchial tree of the rats was 1.587, but that of juveniles was larger than that of the adults. We found a significant negative correlation between age and fractal dimension. This correlation could be considered be misleading because of the difficulty of separating age/body size effects. Nevertheless, because fractal dimensions of the bronchial tree of rabbits and humans are known to be similar, 1.58 and 1.57 respectively, the body size effect may be nil. To our knowledge, this is the first report of ontogenetic changes in the fractal dimension of the bronchial tree in mammals. PMID- 11021309 TI - [Toxic effect of DDT, chlordane and water from the Ignacio Ramirez dam (Mexico), on Daphnia magna (Crustacea: Daphnidae)]. AB - Chlorodiphenylnitrichloroethane (DDT) and chlordane (CLO) are currently used in Mexico to control malaria and termites. From 1990 to 1996 a total of 27 ton of DDT and 508 of CLO were imported. We establish a methodology to determine their environmental impact in a Mexican dam (Ignacio Ramirez). The toxic effect of DDT and CLO were evaluated on the o-demethylase (OD) and acethycholinesterase activities (AchA) of the cladoceran Daphnia magna exposed to different concentrations of the insecticides solved in water from three sites. Their effect on the AchA and OD activities, and so the CL50 were used as exposure bioindicators to determine the more polluted sites. The physicochemical characteristics of water and the biodiversity of the dam test sites were considered. The station near the floodgate has toxicity potential because enzymatic activities were modified. We suggest the use of AchA and OD activities measure in the cladoceran to evaluate the toxicity of a water body polluted by organochlorate insecticides. PMID- 11021310 TI - Anti-inflammatory activity of aqueous extracts of five Costa Rican medicinal plants in Sprague-Dawley rats. AB - The anti-inflammatory properties of Loasa speciosa and Loasa triphylla (Loasaceae), Urtica leptuphylla and Urera baccifera (Urticaceae), and Chaptalia nutans (Asteracene) were studied using the carregeenan induced rat paw edema model. Aqueous extracts of each plant were made according to the ethnobotanical use. The hippocratic assay was made with female rats; the dose used was 500 mg/kg i.p. and the control group received 0.5 ml of n.s.s.. All the animals treated showed hypothermia, and those treated with the extracts of Chaptalia nutans, Urera baccifera and Urtica leptuphylla showed an increased colinergic activity. Acute toxicities of the aqueous extracts were studied in mice an the mean lethal doses ranged between 1.0226 and 1.2022 g/kg. The extracts of Urera baccifera, Chaptalia nutans, Loasa speciosa and Loasa triphylla (500 mg/kg i.p.) showed an anti-inflammatory activity comparable with that of indomethacin. The extracts of U. baccifera and C. nutans, which showed the greatest anti-inflammatory activity, did not show it when used orally (500 mg/kg p.o.). PMID- 11021311 TI - [2 new species of Stilestrongylus (Nematoda: Heligmonellidae) parasites of Peromyscus (Rodentia: Cricetidae) from Mexico]. AB - Stilestrongylus peromysci n. sp. collected from Peromyscus difficilis (Hidalgo state, Mexico), differs from other species in the genus in number of the spines (30) in the synlophe (both sexes) and because the eighth ray arises from the root of the ninth ray; S. hidalguensis n. sp. parasitised Peromyscus sp. and differs from all other congeneric species in the presence of 24 spines in the male synlophe and in the arrangement of the bursal rays (2-2-1 in the right lobe and 2 3 in the left lobe). A key to the species of Stilestrongylus is provided. PMID- 11021312 TI - Pharmacological properties of the repellent secretion of Zonocerus variegatus (Orthoptera: Prygomorphidae). AB - The odours of the whole body and the secretion of Zonocerus variegatus were easily recognised and perceived by human volunteers. However, the secretion odour is not related to the odour of the food plant consumed by the grasshopper. The repellency of Z. variegatus becomes more pronounced in the 6th. and adult instars whose gland lumens contain an appreciable volume of secretion. The secretion odour is so strong that even dilution does not affect its repulsiveness to humans. The secretion had pharmacological properties: it induced contraction in rat (Rattus rattus) stomach smooth muscle preparations and guinea pig (Cavia porcellus) ileum, and induced oedema formation in the rat hind paw. The secretion was not lethal to the animals used in this study, effects were temporary and recovery occurs after a short time. PMID- 11021313 TI - Hypotensive action of an aqueous extract of Pimenta dioica (Myrtaceae) in rats. AB - The intra-venous (i.v.) hypotensive action of the final aqueous fraction of Pimenta dioica was studied in Spontaneously Hypertensive Rats (SHR). The rats were anaesthetized (sodium pentobarbital 50 mg/kg), the trachea, right carotid artery and jugular vein were cannulated for adequate ventilation, direct blood pressure measurement and intra-venous administration of extracts, solutions and drugs. The arterial line was connected to a pressure transducer (Viggo-Spectramed model P23 XL) and a polygraph (Grass model 7H) and monitored continuously during the first five minutes after plant extract administration and then at 5 and 15 minute intervals for one hour. Responses were taken as the maximum pressure changes observed during this period. Increasing doses of the final aqueous fraction were given i.v. to groups of six SHR each. It produced a dose dependent decrease in blood pressure and the ED50 was 45 mg/kg. To discard that the hypotensive effect of the extracts was due to its ionic composition, a solution containing KCl, NaCl, CaCl2 and MgCl2 equivalent to the ion contents present in a dose of 50 mg/kg of total aqueous extract was injected to Sprague-Dawley rats (SDN) using the same method as described above. It did not produce significant changes in blood pressure. Pharmacological antagonistic studies were done injecting either autonomic ganglion, alpha adrenoceptor, beta adrenoceptor and cholinergic receptor blockers prior to extract administration in SHR rats. Atropine, propranolol and phentolamine did not affect the hypotensive effect of the final aqueous fraction. With hexamethonium (autonomic ganglion blocker) the hypotensive response was diminished in a significant way (p < 0.05). The hypotensive action of the final aqueous extract was not mediated through cholinergic, alpha or beta adrenergic receptors. The extract may posses vasorelaxing activity which could not be evident after autonomic ganglion blockade due to extreme vasodilation present prior to extract administration. Future studies should address the question of a possible direct vasodilating effect of the extracts. PMID- 11021314 TI - [Physicochemical and biological characterization of the water quality in the Piedras Blancas basin, Antioquia, Colombia] [In Process Citation] AB - From April to December 1996 a limnological study was carried out in 17 streams near Medellin, in order to stablish their water quality. The area is located in Piedras Blancas Park (6 degrees 8' 20"N, 75 degrees 30' 20"W) SE of Medellin at 2,400 m altitude. Three sampling periods, encompasing wet and dry season were selected: April-May, August-September and October-December. For qualitative collection methods hand screen and D-net were used. Each station was intensively sampled to collect most macroinvertebrates in all types of habitat. The area sampled in each station was 6 m2. Physicochemical variables in general showed small fluctuations along the study; only conductivity and total dissolved solids exhibited significant changes, all related with high precipitation in the wet season. Macroinvertebrate community consisted of 113 genera, 63 families and seven phyla. Trichoptera was the most abundant group. The results indicate that the freshwater ecosystems in the study area present a high water quality corresponding to a oligo-mesotrophic system. PMID- 11021315 TI - [Parasitic helminths from Sigmodon hispidus (Rodentia: Cricetidae) from seasonal and evergreen habitats in Costa Rica]. AB - The helminthological fauna of the cotton rat, Sigmodon hispidus in a tropical environment varies according to habitat and feeding behavior. Six species of nematodes (Longistriata adunca, Trichostrongylus sigmodontis, Strongyloides sigmodontis, Litomosoides carinii, Monodontus sp. and Protospirura sp.) and two species of cestodes (Hymenolepis diminuta and Raillietina sp.) were found in rats from extensive dry lands in Guanacaste where hot temperatures and heterogeneous diet are the rule. Only two species of nematodes (Longistriata adunca and Angiostrongylus costaricensis) were found in rats collected in a humid pineapple plantation in the Central Plateau (Alajuela) where mild temperatures predominate. A. costaricensis, a metastrongylid of medical importance, was found in 42% of them. PMID- 11021316 TI - Histopathological characterization of nephritides in the brown capuchin monkey, Cebus apella (Primates: Cebidae). AB - The renal lesions are of special importance in the captive primates. The most commonly pathologies are: pyelonephritis, nephrocalcinosis, glomerulonephritis, congenital malformations, hydronephrosis and functional diseases. We report the histopathological study of renal lesions of five cases of deaths in Cebus apella (Primates) of the Argentinean Primate Center. The ages of the monkeys were from 4 months to 15 years old. Microscopically, we have observed principally acute diffuse proliferative glomerulonephritis, hilar mesangio proliferative glomerulonephritis, extracapilar glomerulonephritis with crescents, chronic interstitial nephritis and chronic pyelophritis. PMID- 11021317 TI - Parvovirus-mediated antineoplastic activity exploits genome instability. AB - The generation and accumulation of genetic mutations have been associated with carcinogenesis. Epidemiological and experimental evidence implicate parvoviruses in growth inhibition (oncosuppression) and selective destruction (oncolysis) of tumor cells. It is proposed here that parvoviruses can preferentially target genetically unstable tumor cells, which are deficient in DNA repair mechanisms. This selective strategy may serve as a virus-based therapeutic approach against cancer. PMID- 11021319 TI - Established histological identity and cell destruction treatments for cancer. AB - During cancerous cell turnover activity maintained by two types of mitosis, maturation and heteroduplication, cancer tissue consists of two types of cells, maturable and non-maturable. Most of the tissue is composed of maturable cells, which eventually disappear in the terminally matured cell phase. These cells do not participate in cell turnover activity or the organoid identity of cancer tissue. However, a small portion of the tissue is comprised of non-maturable cells, which replicate themselves endlessly, while producing maturable cells through each mitotic division in hetero-duplication mitosis. Thus, cell turnover activity and organoid identity are established in the cancer tissue. This organoid identity is solely responsible for carcinogenesis. Since most typical features of cancer are only detectable in maturable cells during maturation mitosis, cell destruction targeting these features should not be regarded as eradication. To eradicate cancer, the organoid identity of cancer, which is only established by heteroduplication mitosis, should clearly be recognized, and a new concept of cancer treatment based on destruction of the organoid identity should be devised in the future. This does not appear to be an insurmountable task. PMID- 11021318 TI - Mass flux balance-based model and metabolic flux analysis for collagen synthesis in the fibrogenesis process of human liver. AB - A mass flux balance-based stoichiometric model for human liver metabolism has been set up. The model considers 125 reaction fluxes, and there are 83 metabolites that are assumed to be in pseudo-steady state. Theoretical metabolic flux distributions in the fibrotic and healthy liver cells were determined by maximizing respectively the collagen and palmitate synthesis in the objective function for the solution of the model. The flux distribution maps of the analysis for the collagen synthesis showed that the glycolysis pathway was active down to fructose-6-phosphate and the gluconeogenesis pathway was active up to glyceraldehyde 3-phosphate synthesis. However, the flux distribution maps for the palmitate synthesis revealed that both the glycolysis pathway and the gluconeogenesis pathway were active towards 3-phospho glycerate. The TCA cycle operated from citrate towards oxalacetate, and the anaplerotic reactions that connect the TCA cycle to the gluconeogenesis pathway were active in both analyses. Metabolic flux analysis shows that the amino acid fluxes are indeed important in the collagen synthesis. The results of the comparative analyses for the occurrence of the collagen synthesis in the fibrotic liver cells reveal that among the non-essential amino acids three, namely glycine, proline and aspartic acid, and among the essential amino acids one, methionine, are respectively the potential metabolic bottlenecks and the limiting amino acid. The diversions in the pathways and certain metabolic reactions are also presented, and potential strategies for controlling the collagen synthesis and consequently the fibrosis are also discussed. PMID- 11021320 TI - Gout attacks and lunar cycle. AB - The aim was to search for periodical fluctuations in daily occurrence of gout attacks during the synodic lunar cycle. 126 gout attacks were recorded between 1972 and 1994 at known calendar dates. The synodic plexogram, displaying the number of attacks for each day of the cycle, was processed by cosinor regression. A pronounced cycling was found, with highest peaks under the new and full moon (syzygigies). Thus, the maximal occurrence of attacks coincides with the peaking lunisolar tidal effect. Similar relation of attacks to synodic moon was seen in bronchial asthma of children, and a reciprocal one in paroxysmal tachyarrhythmia. Differing pathogenetic backgrounds of these diseases substantiate the observed differences in their putative reactions on the changing cosmogeophysical environment. PMID- 11021321 TI - Nuclear morphology in breast cancer: is pleomorphism an illusion? AB - Nuclear pleomorphism is an underlying theme of cancer histology. Any medical practitioner, not just the pathologist, would have at one time or another in a Pathology practical class, looked at histopathology slides and convinced himself or herself that cancer cells indeed looked more variable in size and shape than nontumorigenic cells. However, data from our nuclear morphometric analysis of cells in breast cancer and adjacent benign tissues appears to suggest that the larger size of the cancer cell nuclei could make any change in nuclear shape and size more noticeable and apparent than their benign counterparts. We hypothesize that the phenomenon of nuclear pleomorphism is due partly to observational subjectivity. PMID- 11021322 TI - A threshold concept for cancer therapy. AB - Oxygen free radicals have been implicated in many disease processes, including aging and carcinogenesis, and have been associated with a variety of complications resulting from the treatment of cancer. As a result, the treatment of free radical-induced disease with antioxidants or free radical scavengers has become an important therapeutic modality. Ironically, these same oxygen free radicals also play a critical role in anti-cancer therapies. The use of antioxidants such as superoxide dismutase (SOD), in this setting, has been found to decrease the efficacy of anti-tumor therapies, which depend on free radical generation for their action. In addition, increased antioxidant activity can often be utilized by the tumor cell to favor increased growth. Therefore, the appropriate application of oxygen free radicals and antioxidants seems to be critically important in designing proper strategies for both prevention and treatment of malignant disorders. This review will summarize free radical and antioxidant regimens that have been employed to date, examine some of the problems associated with these regimens, introduce the 'threshold concept' explaining the dual effects of oxygen free radicals and antioxidants, and discuss a novel hypothesis regarding therapy that could potentially improve outcome in cancer patients. PMID- 11021323 TI - The tensegrity model applied to the lens: a hypothesis for the presence of the fiber cell ball and sockets. AB - The concept of tensegrity refers to the tensional integrity of a system. Such a system mechanically stabilizes itself by balancing the opposing forces of tension and compression. One category of this model involves a 'pre-stressed' condition in which the structural elements are in a state of tensional compression. The other category includes rigid struts that can sustain the opposing forces. The lens of the eye contains epithelial cells that differentiate at the lens equator to fiber cells. With time, these fiber cells are packed into the center of the lens or the lens nucleus. Part of the process of differentiation is the loss of the cellular nucleus and organelles. During maturation of the lens fiber cells, much of the cytoskeleton is lost as a result of proteolysis. As the lens cells mature, ball and socket joints and interlocking ridges appear on the cellular surface. Applying the tensegrity model to the lens, it appears that the lens cells go from prestressed state of tensional compression to the other state that is defined by rigid connections between cells. PMID- 11021324 TI - The prevalence of asthma appears to be inversely related to the incidence of typhoid and tuberculosis: hypothesis to explain the variation in asthma prevalence around the world. AB - The prevalence of asthma varies widely throughout the world. We now believe that asthma is due to airway inflammation caused by an imbalance of the T-lymphocytes in the airway. The atopy that drives asthma is due to a predominance of Th2 lymphocytes in the airway. This paper links the prevalence of asthma inversely to the prevalence of tuberculosis and enteric infection. We will argue that the reason we observe such marked variations in the prevalence of asthma around the world is the fact that in the developing world there is a survival advantage in the fetal immune response to mature from Th2 to Th1 lymphocyte predominant. A Th1 response is required to combat infectious diseases such as typhoid and tuberculosis. Data from the World Health Organization confirms that these two infectious diseases occur very rarely in those communities where the asthma is high. It may be that the clean and infection-free environment of Australia and New Zealand is responsible for the region of Oceania having the dubious honor of having the highest prevalence of asthma in the world. PMID- 11021325 TI - Oxidative stress, acute and regular exercise: are they really harmful in the diabetic patient? AB - Oxidative stress has been involved in the pathogenic process of a variety of diseases including diabetes mellitus. The production of oxidative reactive products has been involved in biochemical changes in bio-molecules that might produce tissue damage directly related to some of the main vascular complications in the diabetic patient. On the other hand, exercise, paradoxically, is a well recognized model of oxidative stress and also an important therapeutic tool in diabetes management. Therefore, the relationship between oxidative stress and exercise in diabetic patients implies an interesting biochemical paradox due to some of the negative effects of exercise principally by the increase of oxidative species in plasma. The effect of oxidative stress during an acute exercise and after an aerobic training period on those patients remains unknown and needs to be studied. PMID- 11021326 TI - Neonatal management at limits of viability: hypothesis based on review of literature and clinical observations. AB - Advances in the surfactant era have resulted in a dramatic rise in the survival of 23-26 week old neonates. For the group as a whole, survival is 50% with a 50% morbidity in survivors. Current literature includes a few 'small baby protocols' suggesting guidelines for management based on institutional experiences. Adopting these with or without any modifications may not be appropriate for other institutions due to the diversity of the patient population involved and the available resources. A review of literature on ELBW neonates reveals an interesting fact that almost all the management strategies and optimal goals in the most critical first few weeks of life involve two numbers--'fifty' and 'eight'! The relevance of these two numbers (or their multiples) in simplifying the management of ELBW neonates is highlighted. PMID- 11021327 TI - The influence of race and gestational age on the age of maximum risk of SIDS in infancy. AB - Risk of sudden infant death syndrome (SIDS) reaches a maximum in the third month. Thereafter, it decreases by half every 40 days or so. It is proposed that the relative sparing of the very young infant is a consequence of an innate (but temporary) characteristic possessed by the newborn infant. Interpretation of available data suggests that this innate characteristic is negatively associated with the infant's level of maturity. This is the basis for the hypothesis that the age at which the risk of SIDS begins to decline at a uniform rate decreases as the infant's gestational age increases. Because of a greater level of maturity at birth, the age at which this occurs in the black infant should be earlier than average. An analysis of data on 32 573 instances of SIDS within the United States between 1985 and 1991 provides support for the hypothesis. PMID- 11021328 TI - The role of brain thyroid hormones in the mechanisms of seasonal changes in mood and behavior. AB - Many individuals experience seasonal changes in mood and behavior. Various theories have been suggested to explain the mechanisms of these changes. However, the mechanisms of seasonal mood and behavioral changes remain unclear. The author suggests that brain thyroid hormones may play an important role in seasonal changes in mood and behavior. This suggestion is based on the facts that seasonal changes in light and temperature may affect the metabolism of brain thyroid hormones and that small alterations of the brain thyroid economy, independent of peripheral changes in thyroid status, may produce significant behavioral effects. The author further suggests that there may be a fault in the thyroid metabolism in the brain in seasonal affective disorder patients, and that fault cannot be identified by studying the peripheral thyroid hormone metabolism. Seasonal mood and behavioral changes may also be related to the interaction between thyroid hormones and different neurotransmitter systems in the brain. PMID- 11021329 TI - Abnormal neuronal ionic flux activity: learning paradigms of schizophrenic thought disorders. AB - New developments in biological psychiatry have prompted the development of our pathophysiological theory of the etiology of schizophrenic thought disorders. We suggest that the presence of brain bionic lesions may produce microneuropathies in certain neuronal electrical pathways, leading to abnormal biological communication. These blockages force the rerouting of incoming electric signals through bypass pathways which divert the message from target to non-target neurons for decoding and integration. Using this model, we were able to link specific schizophrenic disturbances to different types of brain bionic lesion effects. This model should be further investigated to facilitate the understanding of the nature and origin of schizophrenic disorders and to stimulate further interest in this fascinating field. PMID- 11021330 TI - Fast DNA sequencing. AB - A method for fast nucleotide sequencing is described. It is based in the selection of well-known small oligomers able to be hybridized with the unknown target. The selected oligomers are afterwards ordered following a simple statistical approach. The use of capillary electrophoresis for the analysis is emphasized. PMID- 11021331 TI - Induced genome instability as a potential screening test for cancer susceptibility? AB - The variety of mutations associated with carcinogenesis, along with variations in penetrance and environmental factors, complicate the genetic screening for cancer predisposition. It is proposed here that the detection of inherent genome instability as determined by increased mutagen susceptibility may enhance the identification of populations at risk for cancer. In support for this hypothesis, our analysis reveals a strong association between mutagen-induced chromosomal instability in peripheral blood lymphocytes and the propensity for cancers of oral cavity, pharynx, larynx, and lung. DNA instability in response to a variety of mutagens identifies patients with gastrointestinal, brain, endocrine, breast, skin, and hematologic tumors as well as individuals with cancer family syndromes. Induced genome instability therefore appears to be strongly linked to cancer predisposition, and prospective studies may yield a screening test utilizing a panel of mutagens to better identify populations at risk. PMID- 11021332 TI - Sphincter (ring muscle) gymnastics for spinal cord injured and spinal cord transected patients. AB - Sphincter gymnastics seem to be able to recruit central pattern generators- networks of neurons that generate motor patterns--through peripheral rhythmic stimuli and to activate them without supraspinal signals. This physical therapy method is thus suitable for treating spinal cord injured and spinal cord transected patients, for it can reach below the injured segment and feed signals into the central nervous system, thus activating vital organs. This article presents some observations and discusses a possible mechanism. PMID- 11021333 TI - Hyperdynamic circulation in portal hypertension: a comparative model of arterio venous fistula. AB - Complications of portal hypertension remain perplexing physiologic phenomena in the understanding of shunt hemodynamics with multiple theories. Hyperdynamic circulation was also found in sepsis, chronic anemia and arterio-venous (A-V) fistula which relate to an increase in nitric oxide. We hypothesize that portosystemic collaterals may mimic an A-V fistula in which the high-pressure portal blood connects with the lower pressure systemic venous circulation. Although these collaterals decompress the portal circulation, a number of secondary hemodynamic phenomena occur which increase portal blood flow and tend to counteract the portal hypotensive effect of the portosystemic shunt. The consequent increases in cardiac output and portal blood flow perfuse the compromised liver. As portal blood flow increases, collateral flow increases and is nearly totally shunted in the systemic circulation. This shunt may eventually introduce a vicious cycle of hyperdynamic circulation into a compromised host. Ultimately, high-output cardiac failure occurs, leading to cirrhotic cardiomyopathy. PMID- 11021334 TI - Thiamine supplementation to prevent induction of low birth weight by conventional therapy for gestational diabetes mellitus. AB - Conventional treatment for gestational diabetes mellitus increases the proportion of infants born with a low birth weight, a risk factor for cardiovascular disease and diabetes mellitus in later life. Thiamine supplementation during pregnancy may be shown to be a safe preventive measure. During pregnancy, approximately 50% of the women develop a biochemical thiamine deficiency, whereas the thiamine status falls, but remains within normal limits, in most other women. Thiamine is essential for glucose oxidation, insulin production by pancreatic beta-cells and cell growth. It is therefore likely that thiamine supplementation in pregnant women not only improves their glucose tolerance but also stimulates the intra uterine growth, thereby preventing a low birth weight to ensue from conventional therapy which only improves glucose tolerance. PMID- 11021335 TI - A method for reducing the pain and danger of certain arthritic joints. AB - It has been found that by hanging upside-down from the feet, the pain arising from an arthritic hip joint is largely eliminated. If this method is not used, the pain returns rapidly. It is suggested that the direct cause of the pain and damage is the presence of small, hard crystals of apatite or related compounds present in the synovial fluid of the joint. Under the normal pressures involved in standing, walking or running, these damage the joint involved by a process of grinding. If this joint is opened by hanging and swivelling, the crystals are induced to move out of the joint region, thereby alleviating the problem. PMID- 11021336 TI - A psychophysiological model of emotion space. AB - Despite a wide variety of emotions that can be subjectively experienced, the emotion space has consistently revealed a low dimensionality. The search for corresponding somato-visceral response patterns has been only moderately successful. The authors suggest a solution based on an assumed parallelism between emotion coding and color coding. According to the color detection model proposed by Sokolov and co-workers, neurons responsible for color detection are triggered by a combination of excitations in a limited number of input cells. Similarly, a limited number of input channels may feed complex emotion detectors being located on a hypersphere in a four-dimensional emotion space, the three angles of which correspond to emotional tone, intensity, and saturation, in parallel to hue, lightness, and saturation in color perception. The existence of such a four-dimensional emotion space in the subjective domain is shown by using schematic facial expressions as stimuli. A neurophysiological model is provided in which reticular, hypothalamic, and limbic structures constitute input channels of an emotion detecting system, thus acting as the first layer of emotion predetectors. Hypothalamic neurons with differential sensitivity for various transmitters may elicit a subsequent selective activation in a second layer of predetectors at the thalamic level. The latter are suggested to trigger emotion detectors located in cortical areas, the action of which should be revealed by measures of central nervous system activity. Preliminary results from evoked potential studies show that switching between schematic faces that express different emotions may be used as an objective measure for establishing a psychophysiological emotion space. PMID- 11021337 TI - Blocking cholesterol synthesis impairs acquisition of the classically conditioned eyeblink response. AB - Smith-Lemli-Opitz (SLO) syndrome is a congenital disorder characterized by severe mental retardation. Patients with SLO lack 7-dehydrocholesterol (7 dH) reductase, which catalyzes the last step of cholesterol synthesis. Administration of an agent that blocks 7 dH cholesterol reductase, BM 15.766 (BM), leads to a biochemical profile which resembles that of SLO patients, i.e., lower plasma, liver, and brain cholesterol levels accompanied by the appearance of the precursors 7 dH and 8 dH cholesterol. In this article we address the functional consequences of chronic BM treatment on new motor learning by assessing acquisition of the classically conditioned eyeblink response. Just-weaned rats were fed BM by gavage for four months, with half of these rats given exogenous cholesterol during the last two months of BM treatment. Acquisition of the eyeblink response was impaired in BM-treated rats. Impaired acquisition of the eyeblink response was not accompanied by alterations in responsiveness to either the conditioned or unconditioned stimulus. Exogenous cholesterol, a clinically relevant countertreatment, failed to correct for the learning impairment produced by BM treatment. Chronic treatment with a cholesterol synthesis-blocking agent impaired associative learning in just-weaned rats. PMID- 11021338 TI - Pain perception to the cold pressor test during the menstrual cycle in relation to estrogen levels and a comparison with men. AB - Animal and human research has shown that pain sensitivity changes during the menstrual cycle. This has sometimes been ascribed to hormonal variations. The aim of the present study was to examine how perception of pain, induced by the cold pressor test to the dominant hand, was related to gender and phases of the menstrual cycle. A repeated-measures design was used, where twenty-two female students participated at two different phases of the menstrual cycle (days 2-4 and days 20-24). A control group of nineteen male students participated on two occasions, separated by a three week period. The cycle phase during which each woman began her participation was randomized. Pain was induced using the cold pressor test. Pain threshold was determined as the duration of time between when the subject first reported pain and exposure to the painful stimulus. Pain tolerance was determined as the duration of time until the subject withdraw her/his hand from the test water because the pain was too intensive. The results showed that men tolerated significantly greater pain than women. Women's pain threshold was significantly higher during the second phase of the menstrual cycle. Systolic pressure was higher in men than women, increasing more in men in response to cold pressor testing than women. Further research, including measurements of plasma hormone levels during the menstrual cycle, is needed to clarify the role played by estrogens in pain perception. PMID- 11021339 TI - Can an aspect of consciousness be imprinted into an electronic device? AB - Although quantum mechanics allows consciousness to have some effect on the collapse of wave functions, most conventional scientists expect the effect to be quite small. Their experience with expert systems, computer-aided design, etc., would cause them to deny the possibility that a specific human intention could be imprinted into a simple electronic device via a meditative process and that this device could then influence a target experiment in accordance with the specific intention. Here, via two very different target experiments, that prevailing supposition has been experimentally tested and found to be fallacious! For each target experiment, one starts with two identical physical devices, isolates them from each other and "charges" one with the specific intention for the particular experiment. This charging process involved the services of four highly qualified meditators to imprint the device with the specific intention. The devices were then wrapped in aluminum foil and separately shipped, via Federal Express approximately 2,000 miles to a laboratory where the actual target experiments were conducted by others. For the two experiments, the intentions were (1) to decrease (increase) the pH of water by one pH unit and (2) to increase the ATP/ADP ratio in fruit fly larvae so as to significantly decrease their development time. For (1), changes of 0.5 to 1.0 pH units were achieved while, for (2), reductions of approximately 15 percent in larval development time for the imprinted vs. unimprinted device were observed (p<0.005). From a theoretical perspective, (1) a thermodynamic basis is provided for the effect of intention on both the electrochemical potential and flux of molecular species, (2) a conceptual model for linking subtle domains with the physical domain devices is given, and (3) a conceptual model is also given of how such devices may broadcast specific prime directive information via intention-augmented electromagnetism which can "tilt" chemical reactions in appropriate ways within the target experiment. PMID- 11021340 TI - Using administrative data. PMID- 11021341 TI - Using administrative data to reward agency performance: the case of the federal adoption incentive program. AB - This article explores the history of the federal Adoption Incentive Program, the first federal child welfare outcome-based incentive program to rely solely on administrative data, and discusses it within the context of other prior and ongoing federal child welfare incentive programs. Various data-related issues are also examined, including the use of data to project program utilization and costs, set baselines for the number of adoptions, and award funds. Challenges in the legislatively required use of AFCARS data for determining the amount of incentive funds awarded and the impact of the program on AFCARS reporting are also discussed. The article concludes with a look at the impact of the Adoption Incentive Program on adoption and predictions for the future. PMID- 11021343 TI - Improving child welfare performance through supervisory use of client outcome data. AB - Despite their benefits, there is little evidence that outcome data are being widely used by program managers or field level supervisors. Three interdependent factors that facilitate the use of outcome data are well-constructed reports, and organizational culture that supports learning and outcome achievement, and managerial skills in interpreting data and taking relevant action. This article describes an outcome reporting package and training oriented toward frontline supervisors to help them use outcome data, shape a learning culture, interpret data, and take focused action toward improving outcomes for children and families. PMID- 11021342 TI - Measuring contract agency performance with administrative data. AB - This article uses administrative data to analyze the relative performance of contract agencies-those organizations under contract with a city child welfare agency to provide out-of-home care services to children placed in the custody of the public agency-by examining how long it took children placed in out-of-home care to return home to their families. The objective was to determine whether credible empirical evidence could show a relationship between length of stay and the agency providing care. Agency level reunification rates are widely distributed around the mean, indicating that contract agency performance differs and that "agency effects" leave an independent imprint on a child's out-of-home care experience. PMID- 11021344 TI - Data-based organizational change: the use of administrative data to improve child welfare programs and policy. AB - Administrative databases hold the potential to have a significant impact on the development of effective child welfare programs and policies. This article discusses the strengths and weaknesses of administrative databases, issues with their implementation and data analysis, and effective presentation of their data at different levels in child welfare organizations. PMID- 11021345 TI - Costing child protective services staff turnover. AB - This article details the process used in one state to determine the financial costs to the child welfare agency accrued over the course of one year that were directly attributable to CPS staff turnover. The formulas and process for calculating specific cost elements due to separation, replacement and training are provided. The practical considerations inherent in this type of analysis are highlighted, as well as the use of this type of data to inform agency human resource strategies. PMID- 11021346 TI - Assessing the local need for family and child care services: a small area utilization analysis. AB - This article reports the findings of a study to develop a new method for allocating resources to family and child care services within Northern Ireland. Downloads from financial and client databases, together with a survey of social worker case activity, were used to estimate utilization costs across 500 local areas. Regression techniques were then used to account for variations in these costs in terms of local socioeconomic conditions. The resultant needs index represents a fair and equitable means of allocating central family and child care budgets to local units of management and service delivery. PMID- 11021347 TI - Mapping child maltreatment: looking at neighborhoods in a suburban county. AB - This study used administrative data, census data, and geographic information system (GIS) technology to map and analyze the rates and distribution of child physical abuse, sexual abuse, and neglect at the neighborhood level in a suburban jurisdiction. The results provide an example of how administrative data, census data, and GIS technology can give child protective services and community-based agencies a visual tool to help identify and target neighborhoods in which children may be at high risk for maltreatment. PMID- 11021348 TI - Safety, permanency, and in-home services: applying administrative data. AB - This article describes the construction and use of safety and permanency indicators, two aspects of a full set of indicators that also includes child well being and family functioning. The indicators were constructed from Philadelphia's Family and Child Tracking System and were used to examine the city's Services to Children in their Own Home (SCOH) program. Cohort datasets were constructed through the use of extract files, and two independent data file construction algorithms were employed to calibrate the accuracy of the data construction process. The primary unit of analysis was the "family" spell in SCOH services. Contextual variables included family structure, race, and service intensity. The indicators associated with SCOH spells included reports of maltreatment after service, founded maltreatment after service, and out-of-home placement after service. Event history techniques were used to conduct the data analysis. Baseline indicator data for Philadelphia are presented, and future uses for such data are discussed. PMID- 11021349 TI - Using administrative data to assess child safety in out-of-home care. AB - This article describes efforts to produce useful safety measures from administrative data. A measure similar to that proposed by the U.S. Department of Health and Human Services is presented and compared to a measure that takes into account the length of time children are in placement. These measures are also reported for out-of-home care placement types. The challenges posed in constructing such measures from extant data are discussed. PMID- 11021350 TI - Placement stability for children in out-of-home care: a longitudinal analysis. AB - This study examines the number of placement moves experienced over an eight-year period by 5,557 children in one state who first entered out-of-home care between birth and age six. This group comprised 28% of all young children who entered care during this period. Nearly 30% of children in kinship care and 52% of children in nonrelative care experienced placement instability (defined as three or more moves after the first year in care). Children in kinship care, regardless of age, had fewer placement moves than those in nonkinship care. A multivariate analysis found that children who had more than one placement move during their first year of care were more likely to experience placement instability in long term out-of-home care than if they did not move or were moved only once during their first year in care. PMID- 11021351 TI - Data mining in child welfare. AB - Data mining is the sifting through of voluminous data to extract knowledge for decision making. This article illustrates the context, concepts, processes, techniques, and tools of data mining, using statistical and neural network analyses on a dataset concerning employee turnover. The resulting models and their predictive capability, advantages and disadvantages, and implications for decision support are highlighted. PMID- 11021352 TI - Is there a basis for novel pharmacotherapy of autism? AB - No medication has yet been shown to consistently alter the symptoms or the course of autism in the majority of patients. The present pharmacotherapy is mainly palliative and sometimes effective in attenuating specific behaviors. The search for better treatment involves examination of the underlying pathophysiology, the genetic or environmental etiology (including possible iatrogenic causes), and assessment of the clinically-generated evidence of efficacy, including serendipitous or unexplained findings. Subtle neuroanatomic and neurochemical changes are being explored and there are anecdotal reports or limited clinical trials that suggest some therapy might be possible. Secretin is a surprising recent addition to the list of candidates. The pharmacologic mechanism by which these agents might provide such effect is not clear, but hypotheses are beginning to emerge. In addition, the prevention of some uncertain number of autism cases is being investigated by examination of certain vaccinations as putative causative or contributory factors. These topics are reviewed in this article, which has the additional purpose of stimulating novel drug discovery efforts for this enigmatic disorder. PMID- 11021354 TI - Effects of myricetin on glycemia and glycogen metabolism in diabetic rats. AB - In our previous study, we found that myricetin, a naturally occurring bioflavonoid, was able to stimulate glucose transport in rat adipocytes and enhance insulin-stimulated lipogenesis. We report here that after 2 days of treatment with myricetin (3 mg/12 h), hyperglycemia in diabetic rats was reduced by 50% and the hypertriglyceridemia that is often associated with diabetes was normalised. Treatment with myricetin increased hepatic glycogen and glucose-6 phosphate content. It increased hepatic glycogen synthase I activity without having any effect on total glycogen synthase nor phosphorylase a activity. It lowered phosphorylase a activity in the muscle. Thus, the hypoglycemic effect of myricetin is likely to be due to its effect on glycogen metabolism. There was no indication of serious hepatotoxicity with myricetin treatment and therefore, myricetin could be of therapeutic potential in diabetes. PMID- 11021353 TI - Insulin-like growth factor I retards apoptotic signaling induced by ethanol in cardiomyocytes. AB - Apoptosis of cardiac muscle cells may contribute to the development of cardiomyopathy and heart failure. Alcohol (ethanol) abuse is a major cause of cardiomyopathy, but its underlying mechanism remains unknown. To determine whether ethanol causes apoptosis in cardiac muscle and whether insulin-like growth factor I (IGF 1) improves cardiac muscle survival upon ethanol exposure, we have defined the effects of ethanol and IGF I in primary cardiomyocytes. Ethanol decreased cell viability in dose-response manner from 0.2% to 1%. In contrast, ethanol (0.2-1%) did not alter viability of cardiac fibroblasts. To assess the occurrence of apoptosis, DNA fragmentation was determined with quantitation of nucleosomes. Nucleosomes were increased in ethanol-treated cells, thus confirming the apoptotic effects of ethanol. The pro-apoptotic Bax protein and Caspase 3 are important proteins of apoptotic signaling. The content of Bax and the activities of Caspase 3 were increased upon ethanol exposure. IGF I partially suppressed Bax induction, Caspase 3 activation, DNA fragmentation, and increased cardiomyocyte survival. The effects of IGF I on ethanol-induced apoptosis can be inhibited with a chemical inhibitor of PI 3 Kinase (LY-294002), suggesting that anti-apoptotic actions of IGF I involves PI 3 Kinase. These results may have important implications on further understanding the pathogenesis of alcoholic heart disease and the development of new strategies to treat alcoholic cardiomyopathy. PMID- 11021355 TI - Aggravation of ischemia/reperfusion-induced gastric lesions in streptozotocin diabetic rats. AB - We examined the influence of diabetes on ischemia/reperfusion-induced gastric damage in rats, in relation to the antioxidative system. Animals were injected with streptozotocin (STZ: 70 mg/kg, i.p.) and used after 5 weeks of diabetes with blood glucose levels of >350 mg/dl. Gastric mucosal blood flow (GMBF) was measured before, during and after 20 min of ischemia (1.5 ml bleeding per 100 g body weight from the carotid artery) followed by a 15-min reperfusion in the presence of acid (100 mM HCI). At the end of each experiment, gastric damage was observed macroscopically. GMBF was reduced by ischemia in all groups of rats, followed by a gradual return after reperfusion. Ischemia/reperfusion produced hemorrhagic lesions in normal rat stomachs in the presence of 100 mM HCl. These lesions were significantly aggravated when the animals were pretreated with diethyldithiocarbamate, an inhibitor of superoxide dismutase (SOD). Ischemia/reperfusion-induced damage was also markedly exacerbated in STZ-diabetic rats, but this aggravation was significantly suppressed by pretreatment with exogenous SOD or glutathione (GSH). Diabetic rat stomachs showed significantly less SOD activity as well as GSH content than normal rat stomachs. In addition, the deleterious influence of diabetes on the gastric ulcerogenic response to ischemia/reperfusion was significantly mitigated by decreasing the blood glucose levels by daily insulin treatment. These results suggest that the gastric mucosa of diabetic rats is more vulnerable to ischemia/reperfusion-induced injury, and the mechanism may be partly accounted for by impairment of the antioxidative system associated with a reduced SOD activity and GSH content. PMID- 11021356 TI - Genotype analysis of the CYP2C19 gene in HCV-seropositive patients with cirrhosis and hepatocellular carcinoma. AB - This study was conducted to assess whether the genotypic frequency of Smephenytoin 4'-hydroxylase CYP2C19 gene differs in Japanese cirrhotic patients who developed hepatocellular carcinoma. Thirty-eight patients with cirrhosis were studied. The wild-type allele CYP2C19*1 and the two mutated alleles, CYP2C19*2 and CYP2C19*3, were identified by PCR-RFLP method. Individuals with homozygous CYP2C19*2 or CYP2C19*3 mutation and those with CYP2C19*2 and CYP2C19*3 heterozygous mutation were predicted to be the poor metabolizer (PM) phenotype. The overall frequency of PM predicted from the genotyping analysis was 29% (11 of the 38 patients), consisting of 5 patients homozygous for CYP2C19*2, two homozygous for CYP2C19*3 and four heterozygous for the two defects. Among 24 HCV seropositive patients with cirrhosis and hepatocellular carcinoma, the frequency of PM was 41.7% and significantly higher than that observed in 186 healthy controls. We postulate that the PM phenotype caused by the mutation of CYP2C19 gene in cirrhotic patients with HCV infection is associated with a high risk for developing hepatocellular carcinoma. PMID- 11021357 TI - Chroman amide 12P inhibition of lipid peroxidation and protection against learning and memory impairment. AB - Structure modification of the cerebroprotective chroman amide 12 to improve the drug delivery to the target organ by protecting the active hydroxy functional group was carried out in this study. Chroman amide 12P, which the O-acetyl group was served to protect the active group to be delivered to the target organ, was synthesized. Ex vivo antilipid peroxidation activity of 12P was significantly greater than the activity of 12 while the in vitro inhibition of 12P was found to be lower. These indicated that 12P with protected active group effectively reached the brain, the target site, but in vitro, 12P was unable to release its parent or released slowly. Neuropharmacological effect of 12P was investigated in mice. 12 and 12P (50-100 mg/kg, i.p.) showed significant suppression on the hypermotility induced by methamphetamine. 12P (100 mg/kg, i.p.) was more potent than 12, 54.36% and 38.73% suppression, respectively. The result suggested the enhancement of brain delivery and the antagonism against aberrant dopamine release. In the water maze test, 12P (200 mg/kg) as well as tacrine (3 mg/kg) significantly reduced the learning and memory impairment induced by scopolamine (0.5 mg/kg). The results support the enhanced brain delivery and the additional role of radical scavengers in the modulation of brain neurotransmitters in the aberrant condition. PMID- 11021358 TI - Glucuronidation of apomorphine. AB - Apomorphine, a dopaminergic receptor agonist, is largely used in the therapy of Parkinson's disease. In this study, we characterized the glucuronidation of apomorphine and other catechols in rat liver and brain microsomes, using UDP-[U 14C]glucuronic acid and separation of the glucuronides formed by a thin layer chromatographic method. rat liver microsomes glucuronidate apomorphine at a significant rate, that was increased in the presence of dithiothreitol. Two apomorphine glucuronides were separated by high pressure liquid chromatography. We showed by electrospray mass spectrometry that both products were monoglucuronides. Other catechols were also glucuronidated in liver microsomes at various rates, and among them, 4-nitrocatechol was the most efficiently conjugated. in rat brain microsomes, only 4-nitrocatechol was significantly glucuronidated, suggesting that in the liver, several uridine-diphosphate glucuronosyltransferase (UGT) isoforms participate to the conjugation of catechols. To determine which isoforms catalyze apomorphine glucuronidation, two recombinant enzymes expressed in V79 cells were used. The isoform UGT1A6 was unable to glucuronidate apomorphine, but we observed a significant activity catalyzed by the isoform UGT2B1. These results provide, to our knowledge, the first demonstration of apomorphine conjugation by recombinant UGT2B1, and the first evidence of the lack of apomorphine glucuronidation in the rat brain. PMID- 11021359 TI - Glutamate receptors of the AMPA type modulate neurotransmitter release and peristalsis in the guinea-pig isolated colon. AB - To assess the role of AMPA and kainate receptors in modulating neurotransmitter release from the myenteric plexus, the effect of alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA) and kainic acid on endogenous acetylcholine (ACh) and noradrenaline (NA) overflow from the guinea-pig isolated colon was studied. AMPA inhibited spontaneous ACh overflow and increased electrically-evoked NA overflow. Kainic acid did not influence both ACh and NA overflow. AMPA-mediated effects on ACh and NA overflow were significantly reduced by the AMPA/kainate antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, CNQX. The inhibitory effect of AMPA on ACh overflow could be due, at least in part, to the AMPA-induced NA overflow as it was greatly reduced after adrenoceptor blockade and virtually abolished in sympathetically-denervated animals. The possible functional significance of these findings was studied by measuring the efficiency of the peristaltic reflex in the presence of the different agonists. The efficiency of peristalsis was enhanced by AMPA, whereas it was not modified by kainic acid. In conclusion, AMPA receptors, but not kainate receptors, may play a role in the modulation of ACh and NA release and of peristalsis in the guinea-pig colon. PMID- 11021360 TI - Prenatal exposure to low levels of carbon monoxide alters sciatic nerve myelination in rat offspring. AB - Prenatal exposure to low concentrations of carbon monoxide (CO, 75 and 150 ppm from day 0 to day 20 of gestation), resulting in maternal blood HbCO concentrations equivalent to those maintained by human cigarette smokers, leads to subtle myelin alterations in the sciatic nerve of male rat offspring. The rapid growth spurt in pup body weight was related to the period of maximal increase in myelin sheath thickness in both control and CO-exposed animals. A significant reduction in myelin sheath thickness of sciatic nerve fibers, paralleled by changes in the frequency distribution, occurred in both 40- and 90 day-old rats exposed in utero to CO (75 and 150 ppm). Myelin deficit observed in 75 and 150 ppm CO-exposed animals showed up only after the major spurt in myelination but not early during development. The subtle myelin alterations observed in CO-exposed offspring were not accompanied by changes in developmental pattern of axon diameters and did not result in a gross impairment of motor activity. These results suggest that the myelination process is selectively targeted by a prenatal exposure model simulating the CO exposure observed in human cigarette smokers. PMID- 11021361 TI - Correlation between brain glycogen and convulsive state in mice submitted to methionine sulfoximine. AB - It is now well established that in epileptic patients, hypometabolic foci appear during interictal periods. The meaning and the mechanism of such an hypometabolism are as yet unclear. The aim of the present investigation was to look for a putative relationship between glucose metabolism in the brain and the genesis of seizures in mice using administration of the convulsant, methionine sulfoximine. Besides its epileptic action, methionine sulfoximine is a powerful glycogenic agent. We analyzed the epileptogenic and glycogenic effects of methionine sulfoximine in two inbred mouse strains with different susceptibility towards the convulsant. CBA/J mice displayed high response to methionine sulfoximine. The tonic convulsions appeared 5-6 h after MSO administration, without brain glycogen content variations during the preconvulsive period. These mice died of status epilepticus during the first seizure(s). Conversely, C57BL/6J mice displayed low response to MSO. The tonic and clonic seizures appeared 8 to 14 h after MSO administration with only 2% mortality. The seizures were preceded by an increase in brain glycogen content during the preconvulsive period. Moreover, during seizures, C57BL/6J mice were able to mobilize this accumulated brain glycogen, that returned to high value after seizures. The epileptic and glycogenic responses of the parental strains were also observed in mice of the F2 generation. The F2 mice that convulsed early (16%) did not utilize their small increase in brain glycogen content, and resembled CBA/J mice; while the F2 mice that seized tardily (24%) increased their brain glycogen content before convulsion, utilized it during convulsions, and resembled C57BL/6J mice. Sixty percent of the F2 mice presented an intermediate pattern in epileptogenic responses to the convulsant. These data suggest a possible genetic link between the two MSO effects, epileptiform seizures and increase in brain glycogen content. The increase in brain glycogen content and the capability of its mobilization during seizures could delay the seizure's onset and could be considered a "resistance factor" against the seizures. PMID- 11021362 TI - Cardiovascular effects of BRX-005 comparison to bimoclomol. AB - Concentration-dependent effects of BRX-005, the novel heat shock protein coinducer, cardioprotective and vasoprotective agent, on intracellular calcium transients and contractility were studied in Langendorff-perfused guinea pig hearts loaded with the fluorescent calcium indicator dye Fura-2. BRX-005 increased peak left ventricular pressure, the rate of force development and relaxation significantly in a concentration-dependent manner. The amplitude of [Ca2+]i transients was left unaltered by the drug. In contrast to BRX-005, bimoclomol increased both contractility and the amplitude of [Ca2+]i transients. In canine ventricular cardiomyocytes high concentrations of BRX-005 had no effect on depolarization, whereas bimoclomol suppressed action potential upstroke markedly. In guinea pig pulmonary artery preparations precontracted with phenylephrine, BRX-005 induced concentration-dependent relaxation. This effect of BRX-005 was independent of the integrity of endothelium indicating that vasorelaxant effect of the drug develops directly on vascular smooth muscle. PMID- 11021363 TI - Effect of subchronic bimoclomol treatment on vascular responsiveness and heat shock protein production in spontaneously hypertensive rats. AB - This study was designed to investigate the effects of subchronic bimoclomol treatment on endothelial function and expression of 72 kD heat shock protein (HSP 72) in spontaneously hypertensive rats. Endothelial function was tested by monitoring vascular responses to acetylcholine in isolated aortic ring preparations. Polymerase chain reaction was applied to determine HSP-72 mRNA content in the tissue samples. Significant age-dependent declines in relaxation to acetylcholine and vascular HSP-72 mRNA levels were observed in the spontaneously hypertensive animals. Both changes were prevented by subchronic application of bimoclomol suggesting that preservation of endothelial function might be related to sustained levels of HSP-72. PMID- 11021364 TI - Myocardial fiber diameter as a good indicator of outcome in Batista's operation. AB - BACKGROUND: Despite the initial promissory results of partial left ventriculectomy, or Batista's operation, the postoperative mortality associated with the procedure has been too high. We described a histopathologic study performed to identify histological parameters that could help to determine outcomes of patients undergoing this procedure. METHODS AND RESULTS: Myocardial fiber diameter, myocardial fibrosis, thickness of the compact wall, and number of cells presenting from the endocardium to epicardium were analyzed in 32 patients with idiopathic dilated cardiomyopathy who underwent Batista's operation. Data were grouped by patients who died < or = 6 months and patients who survived for > 6 months after the surgical procedure. Additional analyses were performed to compare results according the causes of death and to test the application of these results to biopsy. RESULTS: Myocardial fiber diameter was the only index that could distinguish the two groups. Myocardial fiber diameter < 22 microm distinguished the group of patients who survived the 6-month postoperative period from patients who died during that time with sensitivity of 85.7 and specificity of 72.2. The subendocardial region of the compact wall and the trabecular portion of the wall exhibited comparable results. CONCLUSION: Our results indicate that the myocardial fiber diameter of samples from the trabecular or subendocardial compact wall regions may help predict the outcome of left ventriculectomy. Samples from the trabecular or subendocardial compact wall regions were used for analysis. Further prospective studies involving left ventricular endomyocardial biopsies are necessary to confirm if the use of myocardial fiber diameter in the selection of patients for surgery improves the index of success of Batista's operation. Other factors that are involved remain unclear. PMID- 11021365 TI - Total arterial myocardial revascularization using new composite graft techniques for internal mammary and/or radial arteries conduits. AB - BACKGROUND: Total arterial myocardial revascularization (TAMR) is feasible because of the excellent long-term patency of the arterial conduits. We present five new surgical configurations for TAMR. METHODS: Between December 1998 and July 1999, 34 patients with triple vessel disease underwent TAMR. All patients were in CCS III or IV. Sketelonized internal mammary arteries (IMAs) were used. The surgical techniques for TAMR consisted of Y or T composite grafts constructed between the in situ RIMA and free LIMA graft or radial artery (RA) conduit in three different configurations. Other techniques uses included a T graft constructed between the RA conduit and free LIMA graft in two configurations. Twenty-six (76%) patients underwent contrast-enhanced TTE color Doppler before and after adenosine provocative test, and seven (20%) patients had postoperative coronary angiography. RESULTS: Overall, 144 anastomoses (average number per patient, 4.2) were completed. One (2.9%) patient undergoing an inverted T graft technique died on postoperative day 2. Another patient (2.9%) undergoing the right Y graft technique using IMAs and RA suffered perioperative AMI due to RA conduit vasospasm. Contrast-enhanced TTE color Doppler before and after the adenosine provocative test and at 1 week postoperation revealed a coronary flow reserve (CFR) of 2.1 +/- 0.2 in the LIMA stem, and in the RIMA stem, a CFR of 2.3 +/- 0.3 (P < 0.007). In one patient undergoing the right Y graft technique using IMAs, we found only anomalous flow dynamic parameters of RIMA, suggesting a partial graft closure. The angiographic examination revealed a free LIMA graft closure. At 6 +/- 2.4 months after operation 33 patients were alive and free of angina. The IMAs stem evaluation by TTE color Doppler at follow-up revealed a 2.45 +/- 0.1 mm LIMA diameter and 2.6 +/- 0.2 mm RIMA diameter, which was more than early postoperative data of P < 0.001 and P < 0.007, respectively. CONCLUSION: These data indicate that TAMR in young patients perhaps offers a better postoperative outcome and perhaps should be part of the surgical armamentarium. These techniques apply the "nontouch" principle and should be taken into consideration in patients with a heavily calcified aorta. Contrast enhanced TTE color Doppler is a safe, accurate, and noninvasive test, which allows assessment of IMA patency and CFR evaluation. The flow reserve of the IMAs seems to be adequate for multiple coronary anastomoses. PMID- 11021366 TI - Carotid disease is a risk factor for stroke in coronary bypass operations. AB - The purpose of this study was to investigate the risk of perioperative major cerebrovascular events (MCVEs) in patients undergoing coronary artery bypass grafting (CABG) and to develop preventive therapy. After excluding the patients with marked ascending aortic atheromas and those with combined intracardiac procedures such as valve replacement and aneurysmectomy, 722 consecutive patients who had carotid artery duplex scanning (CADS) and CABG were studied. The results of the study showed the correlation of advanced age, smoking, previous major cerebrovascular event (MCVE), and severe coronary artery disease with high grade carotid artery stenosis of 80-99% (p < 0.05). A total of 13 patients had perioperative MCVE and an analysis of risk factors showed that the MCVE correlated with carotid stenosis of > 60% and reduced cardiac output requiring inotropic support (p < 0.01). Prophylactic carotid endarterectomy (CEA) in patients with 80% to 99% carotid stenosis notably decreased the incidence of MCVE (p < 0.01), and the use of the "pump off" technique instead of standard cardiopulmonary bypass decreased MCVE (p = 0.056). On the basis of these data, prophylactic carotid endarterectomy is effective in averting perioperative MCVE in subjects with carotid stenosis of 80% to 99%, and for those with 60% to 79% carotid stenosis, the pump off technique decreases MCVE. PMID- 11021367 TI - Effect of partial oxygen supply on mitochondrial electron transport system during complete cardiac ischemia. AB - During complete ischemia we assessed myocardial utilization of the small amount of oxygen available. We also determined whether blood cardioplegia has any advantage over crystalloid cardioplegia in this setting. Patients with preserved left ventricular myocardial function and without anterolateral wall infarct or aneurysm were included to the study. Intermittent cold blood and crystalloid cardioplegia were used in 10 patients (group BC) and 9 patients (group CC), respectively. From myocardial biopsies, obtained before and after ischemia, complete electron transport system (ETS) enzyme activities (NDH, SDH, NCCR, SCCR, and COX) and lactate content were analyzed. Biochemical and hemodynamic analyses also were done. Myocardial and blood temperatures were monitored. Ischemic time was longer in group CC (p < 0.05). There were no important differences in biochemical and hemodynamic variables between the two groups. In addition, there was no difference in NDH and SDH activities as well as COX/SCCR and COX/RS-NCCR ratios between the two groups before and after ischemia. After Ischemia, RS-NCCR in group CC and SCCR and COX activities in both groups were lower than the control. For all enzymes, activity change ratios were not different between groups. Myocardial lactate content was increased in both groups after ischemia. However, the increase in group BC was less (p < 0.01). Based on our findings, we believe that the superiority of blood cardioplegia over crystalloid cardioplegia does not depend on oxygen content, but on other factors such as buffering and free oxygen radical scavenger effects among others. However, with the warm and continuous blood cardioplegia technique, oxygen content might be more important. PMID- 11021368 TI - Reduced postoperative atrial fibrillation using multidrug prophylaxis. AB - BACKGROUND: Atrial fibrillation (AFIB) is the most common complication following coronary artery bypass grafting (CABG). Despite three decades of recognition, efforts to reduce the high incidence reported (15%-30%) have been largely unsuccessful. Reasons for postoperative AFIB are likely multifactorial. As a result, we defined a multidrug prophylaxis based on agents known to be individually effective. This method was applied prospectively to a series of consecutive CABG patients with the goal of reducing the incidence of new-onset postoperative AFIB. METHODS: Isolated CABG with cardiopulmonary bypass was performed on 517 consecutive patients. A rapid recovery protocol emphasizing AFIB multidrug prophylaxis was applied to all patients. All patients received 10 microg of triiodothyronine intraoperatively when the clamp on the aorta was released. Immediately following CABG, parenteral magnesium was administered to assure a serum magnesium > 2.2 mEq/dL. Thyroxine 200 microg was administered parenterally to all patients on postoperative days 1 and 2. Metoprolol (25 mg to 100 mg/day) was begun on all patients after extubation provided: heart rate > 85 beats/min and systolic blood pressure > 130 mmHg. Parenteral procainamide (12 mg/kg) loading dose, followed by a maintenance dose (2 mg/min), was used for patients who developed premature atrial contractions (> 1/min), nonsustained supraventricular tachycardia, or any episodes of atrial fibrillation. All patients also received postoperative digitalization, steroids, and aggressive diuresis. RESULTS: The 30-day operative mortality was 3.7%. The overall incidence of new-onset postoperative AFIB was 10.3% (53 patients). There was no major difference in operative mortality (7.5% vs 3.2%, p = 0.23), Parsonnet risk score, or intraoperative variables between AFIB patients and the non-AFIB patients. Patients presenting with a preoperative acute myocardial infarction (p < 0.05), left main stenosis > or = 70% (p < 0.01), and advanced age > or = 70 years (p < 0.05) were at increased risk of developing AFIB. The length of stay for patients with AFIB was 9.9 +/- 9.6 days versus 5.9 +/- 5.2 days (p < 0.001). CONCLUSION: Application of a multidrug prophylaxis can reduce postoperative AFIB to a low incidence. Identification of associated clinical features can help predict patients at risk for postoperative AFIB. Additional strategies to target postoperative AFIB may include treatment at the earliest recognition of atrial rhythm instability. PMID- 11021369 TI - Chronic aortic dissection (DeBakey type II) with infective thrombus in the false channel. AB - A 68-year-old man with a DeBakey type II chronic aortic dissection developed bacteremia secondary to a urinary tract infection with the Serratia species. This resulted in an infected thrombus in the false channel and a subsequent rupture of the ascending aorta. An emergency resection of the ascending aorta and a Dacron graft interposition were performed. The patient suffered from severe neurological insufficiency and died of pneumonia 6 months postoperatively. The optimal operation time and surgical options of an infected aortic dissection should be determined. PMID- 11021370 TI - Sequential revascularization of intramuscular left anterior descending coronary artery. AB - We present a new technique for avoiding possible kinking or angulation of the sequential left internal mammary artery to left anterior descending artery (LIMA LAD) anastomoses when the LAD follows an intramuscular course. A 3- to 5-mm cusp of saphenous vein segment is interposed between the intramuscular LAD segment and internal mammary artery (IMA) at the sequential anastomotic site, to which the distal portion of the IMA was anastomosed in standard end-to-side fashion. PMID- 11021371 TI - Comparison of intermittent warm and cold blood perfusion during hypothermic myocardial preservation on functional and metabolic recovery. AB - Numerous techniques are used to maintain intraoperative heart viability. The studies presented here evaluated heart function and metabolism after various periods of preservation up to 4 hours with intermittent warm and cold blood perfusion. Using a heterotopic heart model cooled to 10 degrees C and maintained for 1, 2, 3, and 4 hours, various preservation techniques were compared. Changes in myocardial metabolism were determined from substrate uptakes and biopsy samples of the left ventricular muscle for high-energy phosphates. Preservation techniques included: (1) sustained hypothermia, (2) 1 or 2 hours of sustained warm blood perfusion with fibrillation, (3) intermittent cold blood perfusion during 2, 3, and 4 hours of preservation, (4) intermittent warm blood perfusion during 2, 3, and 4 hours of preservation and (5) a control group (no preservation). Normothermic fibrillation had no effect on postpreservation functional or metabolic parameters. Sustained hypothermia reduced functional recovery proportional to the length of ischemia. The cold intermittent procedures maintained function and metabolism better than sustained hypothermia, while warm intermittent preservation maintained function and metabolism at control levels throughout the recovery period for all preservation techniques. Changes in ATP mirrored the functional changes. Creatine phosphate (CP) was markedly reduced during heart isolation and preservation and exceeded the control by 100% during reperfusion. For operative procedures of 2 hours or less, functional and metabolic recovery was not affected by the various preservation methods applied. Warm intermittent perfusion during hypothermic preservation offered the best protection for the myocardium. The warming cycles during hypothermia may provide some degree of preconditioning and protect the myocardium during reperfusion. PMID- 11021372 TI - Use of the "single suture" technique to expose the anterior surface of the heart. AB - The "single suture" technique, which consists of placing a suture in the oblique sinus of the posterior pericardium and connecting it to a vaginal tape, is commonly adopted in off-pump coronary artery revascularization to obtain elevation of the heart and coronary artery exposure. This report describes the use of this technique to expose the anterior wall of the heart in the setting of ventricular aneurysm repair. PMID- 11021373 TI - Coronary-coronary bypass using the internal thoracic artery: a sparing procedure of the arterial conduit. AB - BACKGROUND: The internal thoracic artery (ITA) is well known to be the best conduit for coronary artery bypass grafting. However, the bilateral use of ITAs remains limited because in situ right ITAs (RITAs) do not possess an adequate length to be directed to the posterolateral myocardium. We thus considered using free ITAs for conduits between the two segments of the same coronary artery. METHODS: From March 1997 to May 1999, 17 patients underwent coronary-coronary bypass grafting (C-CBG) using free ITAs. Early operative results were analyzed. C CBG was indicated when the right ITA had an inadequate length or when a distal part of the ITA was left unused. RESULTS: No patient died after C-CBG and none have experienced angina since C-CBG (mean follow-up period 27.3 +/- 19.8 months). Postoperative angiography was performed in all subjects at discharge. Only one coronary-coronary bypass graft was occluded, the other grafts were patent, and there were no stenotic changes. Bilateral ITAs were used in 75% of the patients undergoing CABG during the period of this study. CONCLUSIONS: C-CBG can expand the use of bilateral ITAs and can provide an alternative method for revascularization of the posterolateral myocardium. PMID- 11021374 TI - An effective technique to correct anterior mitral leaflet prolapse. AB - Up to one-third of the patients with degenerative mitral valve disease and severe mitral regurgitation have anterior mitral valve prolapse due to chordal rupture or elongation. Surgical treatment of such a condition is often technically demanding and not infrequently associated with suboptimal results. Techniques used to treat anterior leaflet prolapse include chordal transfer, chordal shortening, artificial chordae, and anterior leaflet resection or plication. Each of these strategies has potential shortcomings, and there is considerable controversy concerning the durability of anterior leaflet prolapse repairs using these techniques. The "edge-to-edge" technique, a simple and effective method of correcting anterior mitral leaflet prolapse is described. PMID- 11021375 TI - Surgical techniques for the repair of anterior mitral leaflet prolapse. AB - Myxomatous disease is the leading cause of mitral valve regurgitation in the developed world. Although posterior mitral leaflet (PML) prolapse is the most common cause of regurgitation and of repair, lesions of the anterior mitral leaflet (AML) are often considered beyond the possibilities of repair. The surgical anatomy and pathology of the AML and a precise terminology for the location of the lesions are described. The surgical maneuvers for commissural prolapse, chordal shortening, chordal replacement, leaflet resection, and annuloplasty are also described. The advantages and limitations of each maneuver are discussed. It is concluded that a flexible approach to the repair of the AML is necessary to adapt the best technique for each type of lesion. No particular technique can be applied systematically to all lesions. It is the author's contention that the majority of AML lesions can be repaired safely. PMID- 11021376 TI - A peril of minimally invasive cardiac surgery. PMID- 11021377 TI - The rule of ones and bilateral enucleation. PMID- 11021378 TI - Enucleation: past, present, and future. PMID- 11021379 TI - Evisceration: a new technique of trans-scleral implant placement. AB - PURPOSE: To describe a modification of the evisceration technique. METHOD: Trans scleral implant placement of a hydroxyapatite spherical implant. RESULTS: Forty nine patients received implants ranging in diameter from 16 mm to 20 mm. Minor complications occurred in three patients. The functional and aesthetic result in all patients was excellent. CONCLUSIONS: The described modified evisceration technique has advantages over standard evisceration techniques. PMID- 11021380 TI - Enucleation of blind, painful eyes. AB - PURPOSE: To determine the demographics and results of treatment in patients undergoing enucleation of blind, painful eyes. METHODS: Medical records were reviewed to determine the precipitating cause, preoperative symptoms, prior therapy, and results of treatment in 58 patients in whom enucleation was performed to relieve ocular discomfort. Patient perception of symptoms and treatment was assessed by a follow-up mail survey. RESULTS: Forty-five percent of patients had sustained prior trauma. Ocular discomfort was attributed to phthisis (43%), elevated intraocular pressure (40%), or corneal decompensation (10%). Preoperative symptoms included ocular pain (91%), periocular headache (17%), tearing (26%), and ipsilateral photophobia (26%). Eight patients experienced contralateral photophobia. Enucleation provided complete symptomatic relief in 54 patients (93%). Two of the four patients with persistent discomfort experienced painful facial paresthesias, attributed to prior retrobulbar alcohol injections. Ninety-one percent of the 32 patients who completed the follow-up survey reported pain relief after surgery, whereas 94% would recommend enucleation to others with blind, painful eyes. CONCLUSIONS: Trauma is the most common precipitating condition in patients undergoing enucleation of blind, painful eyes. These patients experience a variety of symptoms, including ocular pain, facial pain or paresthesias, tearing, and photophobia. Contralateral photophobia is not uncommon. Enucleation is highly effective in relieving these symptoms and should be considered in patients unresponsive to topical therapy. Facial paresthesias in patients who have received prior retrobulbar alcohol injections are not relieved by enucleation. PMID- 11021381 TI - Porous polyethylene implant fibrovascularization rate is affected by tissue wrapping, agarose coating, and insertion site. AB - PURPOSE: Often used in facial and ocular reconstruction, biointegratable materials, such as hydroxyapatite and high density porous polyethylene, can be associated with migration, exposure, and infection. Complications are less likely after implants become fibrovascularly integrated. A model was sought to study the influence of multiple factors on the rate of fibrovascular ingrowth into porous implants. METHODS: High density porous polyethylene cubes were implanted into paraspinous skeletal muscles in rabbits. The cubes were explanted at weekly intervals using survival surgery. The number of fibroblasts at the center of each cube was counted, generating a time-dependent standard curve of cell accumulation. Porous polyethylene cubes uncoated, coated with agarose (a plant derived carbohydrate), or coated with nonperforated sclera (human or rabbit) were implanted into suprascapular adipose and paraspinous skeletal muscle in other rabbits. RESULTS: Fibrovascular ingrowth occurred more rapidly with cube implantation into skeletal muscle versus adipose, with increased surface area contact between implants and muscle, and with removal of muscle capsules. While the rate of fibroblast accumulation decreased in cubes coated with sclera, coating the cubes with agarose increased the fibrous capsule formation without altering the rate of biointegration. CONCLUSIONS: This study provides a novel approach for the study of fibrovascular ingrowth into implants treated under a variety of conditions. Modification of current surgical techniques may increase the rate of porous polyethylene implant biointegration. PMID- 11021382 TI - Epidermal and fibroblast growth factors enhance fibrovascular integration of porous polyethylene implants. AB - PURPOSE: Porous implants used in functional and aesthetic reconstruction of the orbit, face, and cranium are less likely to develop complications after they become biointegrated. We investigated whether the administration of exogenous growth factors could increase the rate of implant integration. METHODS: High density porous polyethylene cubes were placed in dorsal paraspinal muscles of rabbits, and daily transcutaneous injections of saline, epidermal growth factor, or basic fibroblast growth factor were administered directly over the cubes for 10 days. At serial time points up to 10 weeks, cubes were explanted and the fibroblasts present at the center of the cubes were counted. RESULTS: Injections of epidermal growth factor and basic fibroblast growth factor increased the rate at which fibroblasts accumulated in porous polyethylene implants and decreased the time required to achieve a maximal rate of cellular accumulation within the cubes. At 4 weeks, when all cell populations had attained a linear rate of accumulation, cubes previously injected with saline, epidermal growth factor, or basic fibroblast growth factor contained an average of 10, 40, and 80 cells per 0.0156 mm2, at their centers, respectively. CONCLUSIONS: Enhancement of the rate of biointegration of porous polyethylene cubes in rabbits is achievable by repeated, transcutaneous administration of exogenous growth factors. PMID- 11021383 TI - Growth factors embedded in an agarose matrix enhance the rate of porous polyethylene implant biointegration. AB - PURPOSE: Repeated injections of epidermal and basic fibroblastic growth factors have been shown to enhance the biointegration rate of implanted porous polyethylene. A study was done to determine whether agarose, introduced at the time of implant placement, might serve as an adequate "single dose" delivery system for endogenous and exogenous growth factors. METHODS: Polyethylene cubes coated with agarose-containing growth factors were implanted into fat and muscle in rabbits. Factors studied included autogenous whole blood, autogenous serum, ascorbic acid, epidermal growth factor, basic fibroblast growth factor, transforming growth factor alpha, and transforming growth factor beta. The rate and character of the fibrovascular ingrowth into implants and surrounding capsule thickness were assessed. RESULTS: Fibroblast infiltration enhanced two- to sixfold with the use of autogenous or allogenic factors introduced in an agarose matrix at the time of cube implantation. CONCLUSIONS: Growth factors studied altered the thickness of the capsule surrounding implants as well as both the vascularity and stromal density within implants. PMID- 11021384 TI - The bioceramic orbital implant: a new generation of porous implants. AB - PURPOSE: The authors describe a new generation of porous orbital implant made of aluminum oxide (Al2O3) and compare it with the hydroxyapatite orbital implants (Bio-Eye and FCI hydroxyapatite). METHODS: The authors examined the new implant macroscopically, with chemical analysis and microscopically with scanning electron microscopy. Animal implantation studies were performed using six adult male New Zealand albino rabbits. Implant vascularization was evaluated by means of magnetic resonance imaging and histopathologic sectioning. RESULTS: The Bioceramic orbital implant was found to have very uniform pore structure with an average pore size of 500 microm. The implant was 99.9% aluminum oxide on x-ray diffraction. Magnetic resonance imaging in vivo vascularization studies demonstrated enhancement of the implant to its center by 4 weeks after implantation in the rabbit. Histopathologically, fibrovascularization occurred uniformly throughout the implant and was noted by 4 weeks. CONCLUSIONS: The Bioceramic orbital implant represents a new porous orbital implant that has a very regular and extensive interconnected pore system, is as biocompatible as hydroxyapatite, is easy to manufacture, structurally strong, and free of contaminants. It is manufactured with no disruption to marine life ecosystems as may occur in the harvesting of coral for other orbital implants. It is less expensive than currently available hydroxyapatite implants and was approved by the U.S. Food and Drug Administration in April 2000. PMID- 11021385 TI - The Molteno M-Sphere. AB - PURPOSE: To analyze a mammalian hydroxyapatite (HA) implant known as the Molteno M-Sphere, recently approved by the Food and Drug Administration of the United States. METHODS: The authors examined the implant macroscopically, with chemical analysis (x-ray powder diffraction, x-ray fluorescence spectrophotometry), and microscopically with scanning electron microscopy. Animal implantation of six Molteno M-Spheres was carried out in six adult male New Zealand albino rabbits. Implant vascularization was evaluated by means of magnetic resonance imaging and histopathologic sectioning. RESULTS: The M-Sphere was found to have multiple interconnected pores throughout with an average pore size of 300 microm to 600 microm. This implant was very light-weight (0.31 g) and fragile. It was made up of pure HA. Magnetic resonance imaging studies showed implant enhancement to its center by 4 weeks after implantation. Histopathologically, fibrovascularization occurred uniformly throughout the 4, 8, and 12-week rabbit implants. CONCLUSIONS: The M-Sphere is an alternative type of HA implant that recently has been reintroduced into the United States for use after enucleation, evisceration, or as a secondary implant. It has multiple interconnected pores allowing central fibrovascularization as early as 4 weeks in a rabbit model. Its light weight and fine trabecular framework, however, are associated with increased implant fragility when compared with other available HA implants (BioEye and FCI3 synthetic HA). The implant requires careful handling because routine handling may damage the implant. The implant is currently approved by the United States Food and Drug Administration. PMID- 11021386 TI - Brazilian hydroxyapatite implant. AB - PURPOSE: To evaluate a synthetic hydroxyapatite (HA) (manufactured in Brazil) in a rabbit model. METHODS: Nine New Zealand white rabbits underwent enucleation of one eye followed by implantation of either a 12-mm Brazilian synthetic HA implant or a 12-mm BioEye, wrapped in polyglactin 910 mesh. Magnetic resonance imaging was performed to assess fibrovascular ingrowth 4, 8, and 12 weeks after implantation. Three animals were killed at each of the times for histopathologic examination. The Brazilian implant was also examined chemically and by scanning electron microscopy. RESULTS: The Brazilian HA was found to be heavier than either the BioEye or synthetic FCI3 HA implants. It did not have a visible, regularly arranged interconnected porous architecture. Rather, it had randomly appearing channels apparent on its external and internal surface in addition to large cystic areas within the body of the implant. Scanning electron microscopy revealed the implant to have a microporous architecture in addition to the large channels and cystic cavities. Histopathologically, central vascularization occurred by 4 weeks and was also present at 8 and 12 weeks. In three Brazilian implants an unrecognizable, nonbirefringent material was identified. The cause of it was unclear. Chemical analysis confirmed the implant to consist of pure HA. CONCLUSION: The Brazilian implant is pure HA that appears solid but has randomly arranged channels as well as a microporous architecture that allows vascularization to its center. It is heavier than other available HA implants and has the presence of an unidentified foreign material within it. Although this implant is less expensive and does not require a costly manufacturing process, the structural characteristics of the material do not offer any theoretical or clinical advantages. The implant is only available in Brazil at this time. PMID- 11021387 TI - Primary placement of a titanium motility post in a porous polyethylene orbital implant: animal model with quantitative assessment of fibrovascular ingrowth and vascular density. AB - PURPOSE: To determine whether a Medpor porous polyethylene orbital implant, at the time of initial orbital implant surgery, will tolerate the insertion of a titanium screw on the anterior surface of the implant. METHODS: Twelve New Zealand white rabbits were enucleated and implanted with a porous polyethylene orbital implant. At the time of enucleation, the porous polyethylene orbital implants were drilled, and titanium motility coupling posts were inserted. The motility coupling posts were inserted at two projection heights (2 or 4 mm) and either covered within Tenon capsule/conjunctiva (eight implants) or left exposed (four implants). Rabbits were killed at 6 or 12 weeks. Clinical tissue tolerance, histologic response to the motility coupling post, and vascular density of the porous polyethylene orbital implant were evaluated. RESULTS: The motility coupling posts were well tolerated, and extrusion or migration of the motility coupling post did not occur. The average percentage cross-sectional area of the implant occupied by fibrovascular tissue at 6 and 12 weeks was 76.3% and 97.5%, respectively. In comparing the vascular density (number of vessels per square millimeter) in the porous polyethylene orbital implant within a 1-mm zone immediately surrounding the motility coupling post, no significant difference between this zone and the vascular density found within its entire corresponding annulus was found at either 6 or 12 weeks. CONCLUSIONS: During the 6- and 12-week observation periods, all implanted motility coupling posts demonstrated favorable tissue tolerance and stable interfaces with surrounding tissues. The extent of fibrovascular tissue ingrowth and vascular density verify that initial screw insertion does not adversely affect the healing process after porous polyethylene orbital implant implantation. Thus, primary placement of the motility coupling post may obviate the need for a secondary surgical procedure. PMID- 11021388 TI - A new titanium peg system for hydroxyapatite orbital implants. AB - PURPOSE: To evaluate a new hydroxyapatite-coated titanium sleeve and titanium peg system for HA orbital implants. METHODS: The authors followed 54 patients receiving an HA-coated titanium sleeve and peg system and analyzed the complications associated with this peg system. The following data were recorded: type of surgery performed, size of implant used, type of HA used, time of pegging, follow-up duration, problems encountered, and treatment. RESULTS: Fifty seven patients received the HA-coated titanium peg and sleeve system. The average duration of follow-up was 15 months (range, 3-30 months). Three patients were lost to follow-up after 1 month. Complications associated with peg placement in 54 patients included: discharge (9.2%), pyogenic granulomas (14.8%), peg falling out during prosthesis removal (9.2%), poor transfer of movement (1.8%), clicking (3.7%), conjunctiva overgrowing peg (1.8%), part of sleeve shaft visible (9.2%), peg drilled on an angle (1.8%), HA visible around peg hole (3.7%), and loose sleeve (3.7%). CONCLUSION: The HA-coated titanium sleeve and titanium peg is a new peg system available for HA orbital implants. Many of the complications associated with this peg system are similar to the commonly used polycarbonate peg system. Pyogenic granulomas and discharge, however, appear to be less frequently encountered with this new system. The HA-coated titanium sleeve and titanium pegs were well tolerated and appeared quieter in the socket than most polycarbonate pegs. PMID- 11021389 TI - The ideal ocular prosthesis: analysis of prosthetic volume. AB - PURPOSE: An ocular prosthesis should complement the volume of the intraconal implant to achieve complete replacement of the volume that is removed by enucleation. This study investigates the limitations on the prosthetic volume in achieving this goal. METHODS: Prosthetic volume and thickness were measured in 70 patients who underwent enucleation. Patients in group 1 (n = 17) were adults enucleated in childhood, and patients in group 2 (n = 53) were enucleated as adults. Clinical problems after enucleation were documented to determine problems potentially related to prosthetic volume. RESULTS: None of the ocular prostheses in this patient series was greater in volume than 4.2 ml (range, 0.75-4.2 ml). The average prosthetic volume for patients with implant diameters of 14 mm to 22 mm was 2.2 ml. A prosthetic volume > or =1.8 ml provided an anterior to posterior dimension of 7 mm. Anterior malposition of the implant and the presence of severe socket contraction were noted in patients with the smallest and the thinnest prostheses. In group 2, larger prostheses were associated with ptosis and lower eyelid laxity (p < 0.05; p < 0.01). CONCLUSIONS: One should not depend on the ocular prosthesis to supply more than 4.2 ml of volume in the anterior compartment of the socket. Adult patients with normal bony development, noncontracted sockets, and an average axial length should not receive implant sizes <20 mm, if one intends to achieve complete replacement of the volume removed by enucleation. Children should receive the largest implant possible. PMID- 11021390 TI - Norian craniofacial repair system bone cement for the repair of craniofacial skeletal defects. AB - PURPOSE: To describe the use of the Norian Craniofacial Repair System (CRS) calcium phosphate bone cement in the restoration of craniofacial skeletal defects. METHODS: Consecutive case series. RESULTS: Calcium phosphate bone cement was used to repair craniofacial skeletal defects in three patients. Indications included repair of a posttraumatic orbital floor defect causing hypo-ophthalmos, reconstruction of frontal craniotomy and temporalis muscle donor sites in a patient who had undergone resection of an invasive squamous cell carcinoma, and augmentation of a post-traumatic anterior maxillary skeletal defect. The primary outcome measure was the restoration of bony volume and support. The use of calcium phosphate bone cement in these patients was effective and without complications. CONCLUSIONS: Norian CRS calcium phosphate bone cement is useful in the repair of craniofacial skeletal defects. PMID- 11021391 TI - Rotavirus infection and intussusception: a view from retrospect. AB - A live orally-administrable rhesus rotavirus (RRV) tetravalent (TV) vaccine, licensed in the U.S.A. and the European Union, was recalled from the market because it was suspected to increase the risk of intussusception during the week following immunization. In contrast, natural rotavirus infection is generally believed not to cause intussusception. Because my experience contributed to the first paper that linked intussusception with rotavirus infection, I have re examined our own data published 22 years ago and other studies on this issue. I also made a case study of adenovirus and intussusception as a paradigm to establish an etiological association of viral infection and intussusception. My hypothesis postulated in this review is that natural infection of susceptible (or predisposed) infants with some rotavirus strains, probably serotype G3 rotaviruses, will result in an appreciable fraction of idiopathic intussusception. Thus, the number of rotavirus-induced intussusception cases may change reflecting the relative frequency of G3 strains, which I believe was much higher in the 70s than during the last two decades. The epidemiological data indicate that the RRV-TV vaccine triggers intussusception at a rate significantly higher than the background incidence rate following the week of vaccination, particularly after the first dose. In contrast, the data do not suggest that the cumulative incidence among the vaccine recipients increases accordingly, implicating that the risk of intussusception attributable to the RRV-TV vaccine may be minimal. PMID- 11021392 TI - Antibacterial properties of Pseudomonas aeruginosa immunotype 1 lipopolysaccharide-specific monoclonal antibody (MAb) in a murine thigh infection model: combined effects of MAb and ceftazidime. AB - A murine monoclonal antibody (MAb) specific for the Pseudomonas aeruginosa immunotype 1 (It-1) lipopolysaccharide (LPS) O-side chain was evaluated in terms of its in vitro bactericidal opsonophagocytic activity and in vivo bacterial killing in a mouse thigh infection model. An immunoglobulin (Ig) G2a MAb Ld3-2F2, specific for It-1 LPS, mediated in vitro complement-dependent opsonophagocytic killing at a concentration of 10 microg/ml. MAb-mediated, complement-dependent killing also occurred in the absence of neutrophils at serum concentrations in excess of 20%. A remarkable synergy was observed in opsonophagocytic assays between MAb Ld3-2F2 (0.5 microg/ml) and ceftazidime (1/4 MIC). The administration of MAb Ld3-2F2 at a level of 1 microg resulted in a significant decrease in the number of bacteria in the thigh muscles of normal mice, while 100 microg of the same MAb was required for one log of reduction in the number of bacteria at the same site in neutropenic mice. The combined therapy with MAb Ld3-2F2 and ceftazidime provided a significant reduction in the density of bacteria in the thigh muscle at 9 hr post-infection in normal and neutropenic mice as compared with those after treatment alone or with no treatment (P< 0.01). These favorable in vitro and in vivo interactions of an LPS-specific IgG MAb and ceftazidime strongly support their potential for use in therapy, combined with an LPS reactive MAb and parenteral antipseudomonas beta-lactam antibiotics in the therapy of systemic Pseudomonas infections in normal and neutropenic hosts. PMID- 11021393 TI - Blood lysate staining, a new microscopic method for diagnosis of fungemia using peripheral blood. AB - We developed a microscopy method for the detection of fungal cells in peripheral blood, termed blood lysate staining, using an approximately 5x5 mm dotted blood lysate. This method was able to detect the emerging fungal pathogen Trichosporon asahii in murine models of systemic fungal infection and fungemia in patients quickly and at minimal cost. Pathogenic yeasts were successfully detected in 6 of 8 blood samples which were taken from feverish immunocompromised patients who were clinically suspected of having fungal infections. Fungal cells were observed as ovoid to elongated, 3x3 to 7x10 microm, and occurred singly, budding, and in short chains and clusters in a periodic acid-Schiff-stained blood smear. The yeast cells were easily distinguished from blood-cell debris by their size, shape and smooth yet rigid outline. PMID- 11021394 TI - Comparison of the oral bacterial flora in saliva from a healthy subject and two periodontitis patients by sequence analysis of 16S rDNA libraries. AB - The oral bacterial flora in the saliva from two patients with periodontitis and from a periodontally healthy subject were compared using a sequence analysis of 16S rDNA libraries without cultivation. 16S rDNAs were amplified from salivary DNA by PCR and cloned. Randomly selected clones were partially sequenced. On the basis of sequence similarities, the clones were classified into several clusters corresponding to the major phylum of the domain Bacteria. The major phylum in the libraries was the low G+C Gram-positive bacteria. There was no clonal sequence affiliated with periodontopathic bacteria in the salivary sample from the healthy subject, while a number of periodontal pathogens such as Campylobacter rectus, Prevotella intermedia, Porphyromonas gingivalis and Treponema socranskii were detected in the salivary samples from the patients with periodontitis. In addition, a number of previously uncharacterized and uncultured microorganisms were recognized. These organisms may have some role in periodontal disease. This study reveals some potential for a molecular-biological technique to analyze the oral microflora associated with periodontal disease, including previously uncharacterized and uncultured microorganisms, without cultivation. PMID- 11021395 TI - Firm adherence of Staphylococcus aureus and Staphylococcus epidermidis to human hair and effect of detergent treatment. AB - Staphylococcus aureus and S. epidermidis are common pathogens in hospitals, and care should be taken not to disseminate these organisms among patients. We have focused on human hair as a source of bacterial contamination. We treated hair with culture solutions of S. aureus and S. epidermidis, and then performed scanning electron microscopy. Bacteria were detected on the surface of the cuticles of the hair, and the attached bacteria were not completely removed even by repeated washing with detergents. These results suggested that hair could be a source of bacterial contamination and indicated the importance of decontamination of hair. PMID- 11021396 TI - The rabies virion-associated 100-kDa polypeptide (VAP100) is a host-derived minor component of the viral envelope. AB - We investigated a minor polypeptide component of 100-kDa detected in the rabies virion (referred to as VAP100) by using a monoclonal antibody (mAb), #16743, which was shown to recognize the SDS-denatured VAP100 antigen by immunoblot analyses. Although the VAP100 antigen was hardly detectable in the cell by usual immunoblot methods with this mAb, we could detect the antigen by a luminescent immunoblot method as well as by immunoprecipitation from the metabolically radiolabeled cell lysates and virions. Fluorescent antibody (FA) staining with mAb #16743 detected the uniformly distributed antigen on the formalin-fixed normal BHK-21 cells, while slight accumulation of the antigen was also seen in the Golgi area when the cells were permeabilized by treatment with Triton X-100 after fixation. Rabies virus infection induced alteration of the behavior of VAP100 to show a spotted distribution pattern in virus-infected cells. Double FA staining with mAb #16743 and rabbit antibody against the rabies virus envelope antigen demonstrated colocalized distribution of the viral envelope antigens and VAP100 in the cell. From these results, we think that VAP100 is a membrane associated component of the cell, and its colocalized distribution with the viral envelope antigens in the cell implicates an intimate association of the VAP100 with viral envelope protein(s) and a reflection of possible involvement in the efficient incorporation of VAP100 into the virion. PMID- 11021398 TI - Identification of amino acids of influenza virus HA responsible for resistance to a fusion inhibitor, Stachyflin. AB - We have recently described a novel hemagglutinin (HA) conformational change inhibitor of human influenza virus, Stachyflin (Yoshimoto et al, Arch. Virol., 144, 1-14, 1999). Stachyflin-resistant variants of human influenza A/WSN/33 (H1N1) virus were isolated in vitro and the nucleotide sequences of their HA genes were determined. The relation of amino acid substitutions and Stachyflin resistance was analyzed with in vitro membrane fusion between HA-expressing cells and octadecylrhodamine (R18)-labelled chick erythrocytes (RBC). The amino acid substitutions, lysine to arginine at position 51 or lysine to glutamic acid at position 121 of the HA2 subunit of the HA protein was enough to confer a Stachyflin-resistant phenotype of HA protein. The molecular mechanism of anti-HA conformational change activity of Stachyflin is discussed. PMID- 11021397 TI - Antiviral substance from silkworm faeces: characterization of its antiviral activity. AB - The antiviral activity of a substance (L4-1) purified from silkworm faeces was examined in an HVJ (Sendai virus)-LLC-MK2 cell system. Its antiviral effect depended on the period of light irradiation and was inhibited by sodium sulfite and anaerobic conditions. These results indicate that the antiviral activity of L4-1 is associated with active oxygen species produced from the substance. SDS polyacrylamide gel electrophoretic analysis showed that viral proteins were damaged by this substance under light irradiation. The results suggest that the antiviral activity is due to damage to viral protein(s) caused by active oxygen species produced from L4-1. PMID- 11021399 TI - Expression of recombinant capsid proteins of chitta virus, a genogroup II Norwalk virus, and development of an ELISA to detect the viral antigen. AB - The second open reading frame (ORF2) gene of the Chitta virus (CHV) was cloned to construct a recombinant baculovirus. The CHV ORF2 is predicted to encode a capsid protein of 535 amino acids (aa). CHV showed a high aa identity in the capsid region with genogroup II Norwalk virus (NV) (65-85%), but a low aa identity with genogroup I NV (44-46%). Phylogenetic analysis of the ORF2 gene demonstrated that CHV is genetically closely related to the Hawaii virus included in genogroup II NV. The recombinant capsid protein of CHV (rCHV) self-assembled to form empty virus-like particles (VLPs) when expressed in insect cells with the recombinant baculovirus. An enzyme-linked immunosorbent assay (ELISA) based on antisera to rCHV was developed to detect CHV antigen in stools. The antigen ELISA appeared to be highly specific to both rCHV and CHV-like strains. In addition, combined use of antigen ELISAs using antibodies against two antigenically distinct recombinant VLPs, the recombinant Chiba virus (rCV) and recombinant Seto virus (rSEV), enabled us to determine the genetic as well as antigenic relationship among these three viruses. PMID- 11021400 TI - Monoclonal antibody against Mycoplasma fermentans-specific aminoglycoglycerolipid. AB - Previously, we reported that Mycoplasma fermentans has specific antigens (phosphocholine-containing glycoglycerolipids: GGPL-I and GGPL-III) and discussed the possibility of their pathogenic role. In this paper, we report the characterization of a monoclonal antibody (MF-III-1) specific to GGPL-III (phosphocholine-containing aminoglycoglycerolipid) using methods of electron microscopy, immunofluorescence cell surface staining, laser scanning microscopy, immunoelectron microscopy, and thin-layer chromatography immunostaining. The MF III-1 antibody specifically recognized M. fermentans attached to the surface of HTLV-I-infected human helper T-cells, and it did not cross-react with other lipids nor with human T-cell antigens. Since MF-III-1 distinguishes GGPL-III from GGPL-I, the binding site may include a serinol (2-amino-1,3-propanediol) residue of GGPL-III. MF-III-1 is useful for the in vitro study of M. fermentans, and may also be useful as a tool for the study of the involvement of M. fermentans in human diseases. PMID- 11021401 TI - Determination of the protective effects of neutralizing anti-hepatitis B virus (HBV) immunoglobulins by epitope mapping with recombinant HBV surface-antigen proteins. AB - Anti-hepatitis B virus (HBV) surface-antigen immunoglobulins prepared from human sera are clinical reagents which have been approved for prophylactic treatment in HBV-exposed persons. The passive immunoprophylaxis with immunoglobulins is meant to cross-link viral particles, which are then further cleared by the host's own immune system. While antibodies specific for both anti-S- and anti-preS proteins have been proved to serve as effective anti-viral agents, so far the fine antigen specificity of clinical immunoglobulin preparations has not been determined. Using recombinant proteins covering the hepatitis B surface antigen, in the present study, the specificity of a commercially available immunoglobulin preparation was determined and immunodominant epitopes were mapped. Here, it is shown that the major reactivity of anti-HBV immunoglobulins is directed against the S-protein, and that no reactivity to the preS2 but a weak binding activity to the preS1 region was detectable. The antigen reactivity within the preS1 region was biased to the C-terminal region, which indicates the presence of a putative B cell epitope. The evaluation of the antigen specificity and determination of novel protective epitopes will provide valuable information for the further development and improvement of prophylactic HBV immunoglobulins. PMID- 11021402 TI - First isolation of Stephanoascus ciferrii from a cat. AB - The present study deals with the first isolation of Stephanoascus ciferrii from a cat. A 2-year-old female Persian cat weighing 2.25 kg was referred to an animal hospital with a chief complaint of otitis externa of the left ear. Microscopic examination of specimen from the left ear disclosed yeast cells. The colony of the clinical isolate was cream-colored, rough, raised and wrinkled. The microscopic examination of the clinical isolate revealed abundant branched and septated mycelia with small ramified chains of oval blastoconidia, variable in size, and arranged alongside the hyphae. Amplification of the isolate DNA with LSU rDNA primers yielded a fragment of about 570 bp, whose nucleotide sequence of the isolate showed 100% similarity to that of Stephanoascus ciferrii in the GenBank database. Therefore, the isolate was identified as Stephanoascus ciferrii, confirming the result of mycological examination by molecular analysis. PMID- 11021403 TI - Heterogeneous post-translational modification of Actinobacillus actinomycetemcomitans fimbrillin. AB - Fresh isolates of Actinobacillus actinomycetemcomitans produce bundle-forming fimbriae. The exact molecular mass of A. actinomycetemcomitans fimbrillin, a structural subunit of fimbriae, was determined by liquid chromatography electrospray ionization mass spectrometry. Three major molecular species with 6,226.0, 6,366.0, and 6,513.0 Da were detected in a purified fimbrial fraction from the strain 310-a. These molecular masses were significantly higher than the molecular weight (5,118 Da) calculated from nucleotide sequence data of the fimbrillin gene, flp, suggesting that the fimbrial peptides were post translationally modified. Modification of the fimbrial peptides was also suggested by an N-terminal amino acid sequence analysis of fimbrillin peptic fragments, with the modified amino acids being due to seven serine or asparagine residues located in the C-terminal region. A periodate oxidation/biotin-hydrazide labeling assay of fimbrillin suggested that it might be glycosylated. PMID- 11021404 TI - Molecular cloning and partial characterization of rat procarboxypeptidase R and carboxypeptidase N. AB - Carboxypeptidase R (EC 3.4.17.20) (CPR) and carboxypeptidase N (EC 3.4.17.3) (CPN) cleave carboxy-terminal arginine or lysine residues from biologically active peptides such as kinins or anaphylatoxins in the circulation thereby regulating their activities. Although CPN is present in a stable active form in plasma, CPR is generated from proCPR, a plasma zymogen, by proteolytic enzymes such as thrombin, thrombin-thrombomodulin complex and plasmin. We have isolated rat proCPR and CPN cDNA clones which can induce enzymatic activities in culture supernatants of the transfected cells. mRNA of proCPR was detected only in rat liver by Northern hybridization and showed hepatocyte-specific expression. Expression of proCPR mRNA was enhanced following LPS injection, indicating that proCPR production is increased under inflammatory conditions. PMID- 11021405 TI - Upregulated expression of Iba1 molecules in the central nervous system of mice in response to neurovirulent influenza A virus infection. AB - The present study deals with the expression of Iba1 molecules, a novel EF-hand Ca2+-binding protein, in the brain after stereotaxic introduction of the neurovirulent WSN strain of influenza A virus into the olfactory bulb of C57BL/6 mice. The virus selectively targeted the paraventricular and anterior olfactory nuclei. Infected neurons appeared as early as at day 3 post infection and degenerated and vanished by day 12. The Iba1 molecule was normally expressed in resting microglia. The overexpression of the Iba1 in microglial cells was detected at day 3 post infection, culminating at day 7 with a morphological activation. Iba1-immunopositive macrophages outnumbered microglia in the paraventricular and anterior olfactory nuclei, where the infected neurons had degenerated. Macrophages totally disappeared by day 12, and the Iba1-expression in microglia was reduced to a normal level by day 35. Lack of perforin predisposed the mice to long-term virus infection of the brain, leading to the prolonged Iba1-overexpression. These results show that the Iba1 is upregulated in the mouse brain in response to influenza virus infection and may play significant roles in the regulation of some immunological and pathophysiological functions of microglia during virus infection. PMID- 11021406 TI - Prevention of childhood lead poisoning. AB - Although past national public health efforts have reduced lead exposure significantly, lead poisoning remains the most common environmental health problem affecting American children. Currently, lead exposure occurs predominantly through ingestion of lead-contaminated household dust and soil in older housing containing lead-based paint; exposure can be increased with housing deterioration or renovation. Environmental prevention efforts focus on improvement in risk assessment, development of housing-based standards for lead based paint hazards, and safe and cost-effective lead hazard remediation techniques. Educational efforts address parental awareness of lead exposure pathways, hygiene, and housekeeping measures to prevent ingestion of dust and soil. Blood lead screening is recommended either universally at ages 1 and 2 years or in a targeted manner where local health departments can document a low prevalence of elevated blood lead levels. Nutritional interventions involve provision of regular meals containing adequate amounts of calcium and iron and supplementation for iron deficiency. Lead chelation should complement environmental, nutritional, and educational interventions, when indicated. Collaboration of multiple federal agencies in a new strategy to eliminate childhood lead poisoning should further prevention efforts. PMID- 11021407 TI - Prevention of asthma morbidity: recent advances. AB - Asthma prevalence has risen substantially in recent decades and is an increasing cause of disability for American children. Concern about the rise in morbidity has led to treatment guidelines and a growing body of clinical research. Recent trials continue to support the role of inhaled corticosteroids as the most effective therapy to control airway inflammation associated with persistent asthma. Growth suppression due to inhaled corticosteroids has also been well documented, although the long-term effects and relative potencies of different agents require further study. Other anti-inflammatory agents such as cromolyn and the new class of leukotriene receptor antagonists have demonstrated benefit in milder patients. Leukotriene receptor antagonists and long-acting beta2-agonists may allow for reduction of inhaled steroid doses. Control of environmental allergens and irritants is essential. New evidence suggests an increasingly important role for allergen immunotherapy. PMID- 11021408 TI - Current concepts in adolescent smoking. AB - This review discusses important research findings regarding adolescent tobacco use reported from April 1999 to March 2000. Although the vast majority of adult smokers began before 1 8 years of age, a significant number of college students seem to be initiating smoking behaviors. Recent literature reviews pointed to cultural as well as neurochemical factors that lead to increased tobacco consumption while creating barriers to quitting. Psychosocial and behavioral correlates of cigarette smoking revealed the role that tobacco use plays in coping with cultural, social, and intrapsychic demands. In addition to understanding why and how adolescents initiate and maintain cigarette smoking, recent studies also attempted to uncover the correlates of quitting behavior. However, recent reports of school-based intervention trials revealed that reproducible, long-term success rates may not be achievable with a single program or approach. Finally, several recent studies explored the role that health care providers play in prevention and intervention, as well as the pitfalls of well meaning office interventions. PMID- 11021409 TI - Corticosteroids in the treatment of gastrointestinal disease. AB - Corticosteroids remain the mainstay of anti-inflammatory and immunosuppressive therapy for many gastrointestinal conditions. We are now starting to understand their mechanism of action and the phenomenon of corticosteroid resistance. Because of the ubiquity of corticosteroid receptors in virtually all cells of the body, side effects of therapy are common and may affect multiple body sites. Newer corticosteroid analogues are being developed to minimize these complications, and concomitant use of other immunomodulatory drugs often facilitates corticosteroid dosage reduction or even withdrawal in chronic inflammatory states. PMID- 11021410 TI - Update in gastrointestinal allergic diseases. AB - Food allergy or hypersensitivity is defined as an adverse reaction to food protein which is immune mediated. Without standard definitions and reliable tests for many forms of allergic disease of the bowel, studies are difficult to interpret. The proceedings of a recent workshop on the classification of adverse immunologic reactions to foods provide the framework for this review. Recent studies have helped define the clinical spectrum and natural history of IgE and non-IgE-mediated food allergy, and provide insight into underlying pathophysiology and dietary management. PMID- 11021411 TI - Helicobacter pylori infection in children: update. AB - Despite its worldwide distribution, the pathogenesis of Helicobacter pylori associated gastroduodenal disease remains poorly understood. What is clear is that H. pylori infection rarely resolves spontaneously and that the chronic gastritis that accompanies infection is typically maintained for the duration of infection. Ultimately, if untreated, this chronic inflammation predisposes a subset of individuals to develop gastric or duodenal ulcers and even gastric cancer. Chronic long-lasting H. pylori infection, particularly when acquired early in childhood, can predispose an individual to a significantly increased risk of developing gastric cancer. These studies were so compelling that the World Health Organization has recently classified H. pylori as a type 1 human carcinogen. Since the discovery of H. pylori less than 20 years ago, this infection has continued to generate considerable interest in the medical and scientific community. As we enter the new millenium, there are now a number of effective treatments for children in whom H. pylori-associated peptic ulcer disease is diagnosed. Although there is now overwhelming evidence to confirm that H. pylori plays an etiologic role in the development of peptic ulcer disease, only a small number of these children develop H. pylori disease. In this review, we highlight some of the recently published pediatric studies addressing the role H. pylori plays in the development of gastroduodenal disease in children. PMID- 11021412 TI - New developments in the pathophysiology, clinical spectrum, and diagnosis of disorders of fatty acid oxidation. AB - Fatty acid oxidation disorders are among the most common inborn errors of metabolism affecting infants and children. Recognition of this family of defects is critical because careful dietary monitoring, avoidance of fasting, and prompt intervention during common childhood illness can prevent catastrophic cardiac and metabolic decompensation. This review focuses on new molecular and clinical diagnostic aspects of several of these disorders. Recent papers highlight the recognition that the clinical spectrum of disorders of fatty acid oxidation goes far beyond the stereotypical Reyes-like presentation or cardiomyopathy, and now encompasses more cases of sudden infant death syndrome, fulminant hepatic failure, and severe complications during pregnancy. PMID- 11021413 TI - Infant feeding: a critical look at infant formulas. AB - Commercially available infant formulas serve as the best alternative to human milk when breastfeeding is not possible. Infant formulas are designed specifically to mimic the composition of human milk or the functional aspects of human milk feeding. This review highlights the issues related to the composition of infant formulas. The most hotly debated issue currently is whether to add long chain polyunsaturated fatty acids to infant formulas. Other controversial topics include the safety and efficacy of soy-based protein formulas, protein quantity and quality as they relate to the infant's nutritional needs and feeding tolerance, and the replacement of lactose with other carbohydrate sources for specialized infant formulas. Recent modifications in the fat blend of infant formulas have led to improved fat digestibility. However, the full spectrum of benefits associated with the addition of nucleotides awaits further study. Modifications to infant formulas are made when the preponderance of scientific evidence suggests that the compositional change will better meet the nutritional needs of the infant. PMID- 11021414 TI - Probiotics in pediatric gastrointestinal disorders. AB - Probiotics have been defined most recently as living microorganisms which, upon ingestion in certain numbers, exact health benefits beyond inherent general nutrition. They have been a part of human nutrition for centuries, but in recent years they have been more closely studied for their potential to improve health and treat disease. This review of probiotics is not extensive, highlighting the most recent reviews and well controlled clinical studies in both animals and humans. The safety issues are also discussed as well as potential mechanisms of action. The importance of studying each probiotic bacterium individually in each condition where a health benefit is claimed is highlighted by Lactobacillus GG, the most widely studied probiotic which has proven benefit in reducing the severity and duration of viral diarrhea but no benefit against bacterial diarrhea. PMID- 11021415 TI - Clinical and basic laboratory assessment of children for possible congenital heart disease. AB - Accurate and cost-effective detection of congenital heart disease is a priority for the primary care physician, but there are many diagnostic modalities and strategies available. Within the past year, investigators have reported factors that contribute to the failure to diagnose congenital heart disease before autopsy. Recent research also highlights the strengths and weaknesses of the clinical evaluation by the primary care physician. An examination by a pediatric cardiologist is a more specific discriminator between heart disease and innocent murmur and can, in many instances, eliminate the expense of relying on echocardiography for diagnosis. Current reports document substantial limitations of diagnostic utility of electrocardiography and chest roentgenography. Suspicions that echocardiography can be misleading when performed in laboratories without expertise in congenital heart disease are confirmed in recent organized comparisons of accuracy. High-quality, well-designed tele-echocardiography programs are shown to be effective solutions when maldistribution of pediatric cardiology services hampers diagnostic efficiency. PMID- 11021416 TI - Fetal cardiology. AB - The prevalence of congenital heart disease increases with increasing NT, and many defects can be detected by echocardiography performed by a specialist at 13 to 17 weeks. There is increasing evidence that a prenatal diagnosis improves fetal outcome. Physiology and pathophysiology of the human fetal pulmonary circulation play an important role in the fetal circulation, and recent studies about the reactivity of these vessels in the third trimester have contributed to increase our understanding. The main issue in fetal arrhythmias (brady- and tachycardia) is the correct indication, efficacy, and safety of treatment. PMID- 11021417 TI - Advances in cardiovascular genetics and embryology: role of transcription factors in congenital heart disease. AB - In spite of tremendous advances in diagnosis and treatment of cardiovascular disease in the young, our understanding of its cause is still incomplete. Recent studies have reported pleiotropic cardiac malformations resulting from mutations in transcription factors, a family of proteins known to play important roles in many aspects of development. Further evaluation of these important causes of cardiovascular disease in the young promises new insight into embryology of cardiac development and improved understanding of the pathophysiologic basis of cardiovascular disease in the young. PMID- 11021418 TI - Advances in pediatric cardiovascular surgery: anatomic reconstruction of the left ventricular outflow tract in transposition of the great arteries with pulmonic valve abnormalities. AB - In the past years, advances in pediatric cardiovascular surgery have occurred in many areas with some of the greatest strides being made in complex repairs in younger age groups. Aggressive early corrections while higher risk, may in the long run provide a child with a normal anatomic heart, and corresponding myocardial growth and physiology. PMID- 11021419 TI - Congenital chylothorax. PMID- 11021420 TI - Atopic diseases and upper respiratory infections. AB - This article reviews information on the topics of asthma, atopic dermatitis, food allergy, and upper respiratory infections. The asthma section provides an in depth look at sociodemographic factors contributing to asthma morbidity and the barriers to asthma control. New findings on the triggers and therapies of atopic dermatitis and new articles on formula allergy and peanut allergy are presented. Recent publications in the areas of sinusitis and upper respiratory infections are also reviewed. PMID- 11021421 TI - Bibliography. Current world literature. General pediatrics. PMID- 11021422 TI - Bibliography. Current world literature. Gastroenterology and nutrition. PMID- 11021423 TI - Bibliography. Current world literature. Cardiovascular medicine. PMID- 11021424 TI - Diagnosis of persistent bovine viral diarrhea virus infection by immunohistochemical staining of formalin-fixed skin biopsy specimens. AB - The objective of this study was to evaluate the efficacy of immunohistochemical (IHC) staining for diagnosis of persistent bovine viral diarrhea virus (BVDV) infection using formalin-fixed, paraffin-embedded skin biopsy specimens. Skin from 41 of 42 calves shown to be persistently infected (PI) with BVDV by repeated virus isolation more than 3 weeks apart were immunohistochemically positive for BVDV antigen. Positive IHC staining was most pronounced in the keratinocytes and in hair follicle epithelium, hair matrix cells of the hair bulb, and the dermal papilla. All of the skin sections from 10 calves experimentally infected postnatally with BVDV (10(5) median tissue culture infective doses [TCID50]) and biopsied on days 0, 5, 7, and 9 postinfection were negative for viral antigen. Ten calves from a second group experimentally infected with a higher dose of BVDV (10(8) TCID50) were biopsied when viremic between 10 and 14 days postinfection and 4 calves exhibited positive IHC staining for BVDV; however, staining in these skin biopsies was confined to small foci in the nonfollicular epidermis and follicular ostia. This staining was distinct from that observed in skin obtained from PI cattle. Skin biopsy represents an effective method for identifying animals PI with BVDV. PMID- 11021425 TI - Development and application of a competitive enzyme-linked immunosorbent assay for the detection of serum antibodies to porcine circovirus type 2. AB - We report the development of a competitive enzyme-linked immunosorbent assay (c ELISA) for the detection of antibodies to porcine circovirus type 2 (PCV2), the agent associated with the recently described postweaning multisystemic wasting syndrome in pigs. At present, no method has been published describing a c-ELISA for the detection of antibodies to PCV2, and currently employed tests are impractical for use in some laboratories. The assay described here uses a cell culture isolate of porcine circovirus type 2 as antigen and a PCV2-specific monoclonal antibody as the competing reagent. Evaluation of the ELISA was performed by comparison with results obtained using an indirect immunofluorescent test on 484 sera from pig herds in the United Kingdom, Canada, France, and the USA and serial bleeds from pigs experimentally infected with porcine circoviruses. The sensitivity and specificity of the ELISA were determined as 99.58% and 97.14%, respectively, at 2 standard deviations (SD) from the mean or 95.81% and 100% at 3 SD from the mean. Using this ELISA, a serologic survey of 461 sera collected from commercial pig herds in Northern Ireland between 1973 and 1999 was undertaken. Analysis of the results of this survey demonstrated that the number of ELISA-positive sera detected in an individual year during this period ranged from 55% to 100%. This c-ELISA has applications for large-scale rapid diagnosis of PCV2 infection in pig populations worldwide and for immunoscreening of sera from other species for antibodies to PCV2. PMID- 11021427 TI - The effect of fecal sample weight on detection of Salmonella enterica in swine feces. AB - The effect of different fecal sample weights on the detection of Salmonella enterica in swine feces was examined. Sample weights evaluated were rectal swabs and fecal samples weighing 1 g, 10 g, and 25 g. Comparisons were made on matched fecal samples obtained from individual pigs housed on 2 commercial swine farms in North Carolina. Relative sensitivity (number of positive pigs per fecal weight category/number positive in all weight categories) increased (P < 0.001) with fecal sample weight, and ranged from 9% for rectal swabs to 78% for 25-g samples. Stomaching of fecal samples did not affect detection of S. enterica. These observations demonstrate that fecal sample weight can markedly influence estimates of prevalence of S. enterica in epidemiologic studies. Failure to consider the imperfect sensitivity of bacterial culture in the design and interpretation of epidemiologic studies will lead to underestimation of prevalence and reduced power to detect the presence of S. enterica-infected herds. PMID- 11021426 TI - Malignant catarrhal fever: polymerase chain reaction survey for ovine herpesvirus 2 and other persistent herpesvirus and retrovirus infections of dairy cattle and bison. AB - Using a polymerase chain reaction (PCR) test for sequences of ovine herpesvirus 2 (OHV2), this virus was shown to be significantly associated with sheep-associated malignant catarrhal fever (SA-MCF) in terminal cases of disease in 34 cattle and 53 bison. Ovine herpesvirus 2 was not detected in cattle (38) and bison (10) that succumbed to other diseases. Other persistent herpesviruses, retroviruses, and pestivirus, some of which have been previously isolated from cases of SA-MCF, were not associated with the disease. These included bovine herpesvirus 4 (BHV4), bovine lymphotrophic herpesvirus (BLHV), bovine syncytial virus (BSV, also known as bovine spumavirus), bovine immunodeficiency virus (BIV), and bovine viral diarrhea virus (BVDV). A PCR survey for OHV2 in DNA from individual cow's peripheral blood lymphocytes in 4 dairies showed that the 1 dairy that was in close contact to sheep had a prevalence of OHV2 of 21.3%, whereas the 3 other dairies had no OHV2. Prevalence of the other herpesviruses and retroviruses in the dairy cows was variable, ranging from 2% to 51% for BHV4, 52% to 78.7% for BLHV, and 10% to 34% for BSV. Bovine lymphotrophic herpesvirus and BSV were also found in a few (1-4 of 21 tested) cases of terminal SA-MCF, but BIV and BVDV were not found in either the dairy cows sampled, or in the cases of SA-MCE No significant correlation was found between the presence of any 2 viruses (OHV2, BHV4, BLHV, BSV) in the dairy cows or terminal cases of SA-MCE PMID- 11021428 TI - Coxiella burnetii infection is associated with placentitis in cases of bovine abortion. AB - A positive score on a modified acid-fast (MAF)-stained smear test of fresh placenta was used to identify a group of bovine abortion submissions believed to be infected with Coxiella burnetii. Immunohistochemical (IHC) testing for Coxiella and Chlamydia antigens was performed on 14 MAF smear-positive cases as well as 29 MAF smear-negative cases received during the study period. Nine MAF smear-positive cases as well as 1 MAF smear-negative case were Coxiella-positive via the IHC test. No placentas were positive for Chlamydia antigen. Various histopathologic features were categorized for all placentas and the presence or absence of selected risk categories was also graded for each case. The results between Coxiella IHC-positive cases and Coxiella IHC-negative/MAF-negative cases were compared using Fisher's exact test (P value at 95% confidence). Significant associations were found between Coxiella IHC-positive cases and the presence of placental inflammation (P = 0.0027), placental necrosis (P = 0.012), fetal pneumonia (P = 0.0152), and the visibility of Coxiella-like organisms within trophoblasts on hematoxylin and eosin-stained sections (P < 0.0001). Histopathologic features of Coxiella IHC-positive placentas included infiltration of the chorionic stroma by mononuclear cells, necrosis of chorionic trophoblasts, and focal exudation of fibrin and neutrophils. The results indicate that MAF smears are a good screening tool for the presence of Coxiella in placentas from bovine abortion cases and that the detection of this pathogen in aborted placentas via traditional staining or IHC methods is usually associated with placentitis. PMID- 11021429 TI - The postmortem diagnosis of cholecalciferol toxicosis: a novel approach and differentiation from ethylene glycol toxicosis. AB - The objectives of this study were to develop a novel approach to postmortem diagnosis of cholecalciferol (CCF) toxicosis in dogs using kidney, bile, and urine samples, and to differentiate CCF from ethylene glycol (EG) toxicosis. To achieve these objectives, specimens collected from 2 previous laboratory studies in which dogs were given a single oral toxic dose of CCF (8.0 mg/kg) were used. For EG toxicosis, historical data from the previous 13 years (1985-1998) were reviewed and confirmed cases of EG toxicosis were selected. The historical data were used to compare trace mineral concentrations, specifically of calcium and phosphorus to differentiate between intoxications caused by CCF from that caused by EG in dogs. Kidneys, bile, and urine from dogs that died of CCF toxicosis were analyzed for 25 monohydroxy vitamin D3 (25(OH)D3) and 1,25 dihydroxy vitamin D3 (1,25(OH)2D3) and compared to known control unexposed dogs. Results of this study show that biliary and renal 25(OH)D3 concentrations and renal calcium to phosphorus ratio are of diagnostic value in dogs exposed to toxic concentrations of CCF. The renal calcium to phosphorus ratio was <0.1 in normal dogs, 0.4-0.9 in dogs that died of CCF toxicosis, and >2.5 in dogs that died of EG toxicosis. PMID- 11021430 TI - Activated partial thromboplastin time as a screening test of minor or moderate coagulation factor deficiencies for canine plasma: sensitivity of different commercial reagents. AB - To determine the sensitivity for detection of coagulation factor deficiencies by commercial reagents for canine plasma, 5 commercial activated partial thromboplastin time (APTT) reagents with different types of contact activator and phospholipid of various origin were examined. Thirty canine plasma samples with minor or moderate deficiencies of coagualition factors that influence the APTT were examined. Significant differences were found for the sensitivity of various reagents, but no correlation was found with the type of contact activator. Following the test instructions provided by the manufacturers, the number of APTT results that were prolonged beyond the reference range varied between 20 and 30 (sensitivity = 0.67-1.00); the number of corresponding results using a standardized test protocol varied between 19 and 28 (sensitivity: 0.63-0.93). The most sensitive reagent contained kaolin as a contact activator and a human placental thromboplastin. The results of this study indicate that the APTT test optimized for human plasma is also a sensitive screening test of the intrinsic system of canine plasma, provided that a suitable reagent is used. PMID- 11021431 TI - Bronchoalveolar lavage cytology and hematology: a comparison between high and low health status pigs at three different ages. AB - Bronchoalveolar lavages (BALs) were performed with a bronchoscope on 5- and 7.5 week-old, anesthetized, high health status pigs (n = 14). At 10 weeks of age, pigs (n = 28) were necropsied, lungs were removed, and BAL samples were collected from the right diaphragmatic lobe with a modified 12-Fr (4-mm) Foley catheter. Peripheral blood was sampled from all pigs (n = 28) before each BAL procedure. Peripheral blood and BAL samples were collected according to a similar study design at 5, 7.5, and 10 weeks of age from 12 low health status pigs, which were raised according to standard farm procedures (n = 6) or as segregated early weaned pigs (n = 6). Bronchoalveolar lavage cytology and hematologic 95% confidence intervals were determined for 5-, 7.5-, and 10-week-old high (group A) and low health status pigs (groups B and C). The results were compared between the different groups. Repeated BALs were easily performed in all pigs, making this an additional tool for evaluation of respiratory health. Total numbers of cells and neutrophils in peripheral blood and BAL samples were greater in low health status pigs than in high health status pigs. Hematologic results paralleled the findings in BAL fluid. Segregated early weaning of low health status pigs in a less challenging environment mainly reduced the number of neutrophils in BAL samples and peripheral blood. PMID- 11021432 TI - Atypical Actinobacillus pleuropneumoniae isolates that share antigenic determinants with both serotypes 1 and 7. AB - In the present study, the characterization of 3 atypical isolates of Actinobacillus pleuropneumoniae is presented. Two isolates (1B and 27E) showed positive reactions in coagglutination, immunodiffusion, and indirect hemagglutination tests for serotypes 1 and 7, whereas the third isolate (26B) reacted with antisera to serotypes 1, 4, and 7. These atypical isolates of A. pleuropneumoniae possessed a capsular polysaccharide (CPS) antigenically related to serotype 1 as well as an O-chain lipopolysaccharide antigenically related to serotype 7 or to serotypes 4 and 7, as shown by the use of monoclonal antibodies. Results of toxin profile and virulence assays for mice and pigs showed them to be more related to A. pleuropneumoniae serotype 7 field isolates. All 3 isolates induced antibodies mainly against serotype 7/4 O-long-chain lipopolysaccharide (LC-LPS) and, to a lesser extent, to the CPS of serotype 1, in experimentally infected pigs. Diagnostic laboratories that use a LC-LPS-based enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of A. pleuropneumoniae infection in swine would probably diagnose herds infected with these atypical isolates as being infected by A. pleuropneumoniae serotypes 7 or 4, whereas those that use a CPS-based ELISA would probably consider them as infected by A. pleuropneumoniae serotype 1. PMID- 11021433 TI - Isolation of environmental Clostridium difficile from a veterinary teaching hospital. AB - An environmental survey of a veterinary teaching hospital for the presence of Clostridium difficile was performed using contact plates and cycloserine cefoxitin-fructose with 0.1% sodium taurocholate agar. Clostridium difficile was isolated from 24 of 381 sites (6.3%). Growth was obtained from 4.5% (9/202) of sites sampled in the Large Animal Clinic, from 8.1% (13/160) of sites within the Small Animal Clinic, and from 20% (2/10) of sites sampled elsewhere. Fourteen of 21 strains tested produced toxins in vitro. A geographic association was found with areas in the large animal clinic where nosocomial C. difficile diarrhea in horses had previously been diagnosed. Several other sites with a potential for nosocomial transmission of the organism were identified. Areas from which C. difficile was isolated tended to be areas with high animal traffic, with increased chance of fecal contamination, and with rough, difficult to clean surfaces. This study documents the prevalence of this organism in the environment and its potential role in nosocomial disease. PMID- 11021434 TI - Fatal Clostridium botulinum toxicosis in eleven Holstein cattle fed round bale barley haylage. AB - Twenty-two lactating Holstein cattle in Tennessee had clinical signs of intoxication with preformed Clostridium botulinum toxin. These signs included weakness, paralysis of the tongue and chest muscles, abdominal breathing, and, in 11 of the 22 cows, death. Differential diagnoses included hypocalcemia, hypomagnesemia, carbohydrate overload, and several toxicoses including mycotoxin, lead, nitrate, organophosphate, atropine or atropine-like alkaloid, and botulism. A diagnosis of botulism by the ingestion of preformed C. botulinum type B toxin was made by eliminating these other diseases, by finding C. botulinum type B spores in 3 bales of round bale barley haylage fed to these cattle, and by isolating preformed type B toxin from 1 of the 3 bales. Confirmation of the toxin type was made by demonstrating mouse lethality by intraperitoneal injection of specimen extracts with neutralization by C. botulinum type B antitoxin. The haylage, harvested green and encased in black plastic bags to facilitate fermentation, was presumably contaminated by the botulinum toxin when fermentation failed to produce enough acid to lower the pH to 4.5, the pH below which C. botulinum growth is inhibited. Farmers and ranchers who use round hay balers to produce haylage should be alert to this potential problem. PMID- 11021435 TI - Efficacy of a modified polymerase chain reaction assay for detection of Ehrlichia canis infection. AB - Detection of Ehrlichia canis in acutely infected and convalescent dogs is important for effective treatment and control. However, accurate detection has been difficult to achieve, in part because dogs that have been treated therapeutically often remain seropositive for extended periods. A new method, polymerase chain reaction (PCR) assay using biotinylated E. canis-specific primers (PCR-BP), was developed for detection of E. canis. Four dogs experimentally infected with E. canis by intravenous inoculation of whole blood from carrier dogs and 2 naturally infected convalescent carriers were used to compare the specificity and sensitivity of the new method with that of microscopy/blood smear evaluation, serologic test, and conventional PCR assay using E. canis-specific primers. In experimentally infected animals, infection was detected as early as 7 days post-exposure using PCR-BP. Although the 2 naturally infected dogs were positive by serologic test and PCR-BP, both were negative by conventional PCR. Results suggest that the new method is a sensitive assay for detection of E. canis infection. In addition, results were obtained more rapidly than with other PCR-based assays. PMID- 11021436 TI - Disseminated T-cell lymphoma in a guinea pig with bilateral ocular involvement. AB - A 2-year-old female shorthair guinea pig was presented to the Veterinary Medical Teaching Hospital, University of Wisconsin-Madison, for evaluation of a unilateral corneal opacity of 1 week duration. Physical examination revealed a markedly thickened right cornea and lymphadenopathy of the submandibular and prescapular lymph nodes. Cytology of a lymph node aspirate was highly suggestive of lymphoma. The animal was humanely euthanized. Postmortem examination revealed a disseminated lymphadenopathy involving the submandibular, anterior cervical, prescapular, bronchial, anterior mediastinal, and mesenteric nodes, and hepatomegaly with accentuation of lobular morphology. The right cornea was dark red, dry and dull, and diffusely thickened, and the globe was exophthalmic. Microscopically, pleomorphic neoplastic lymphoblasts were present in the lymph nodes, spleen, liver, lungs, heart, rhinarium, bone marrow, and kidneys. Bilateral infiltration of the eyes by neoplastic lymphoblasts was noted, which was more extensive on the right. The neoplastic cells stained immunohistochemically as T-lymphocytes using antibodies directed against CD3 antigen. PMID- 11021437 TI - Hepatic capillariasis (Capillaria hepatica) in porcupines (Erethizon dorsatum) in Pennsylvania. AB - Tissues of 53 adult porcupines (Erethizon dorsatum) from Pennsylvania were obtained for histopathologic examination. Hepatic capillariasis was recorded in 9% of the porcupines. An additional 11% of the liver sections showed lesions that were compatible with migration by Capillaria hepatica. Because only 1 section of the liver per animal was examined microscopically, the documented prevalence of C. hepatica in Pennsylvania is considered conservatively low. However, this condition was subclinical, because none of the infected animals showed clinical signs, and none revealed severe pathologic changes in the affected livers. This seems to be the only report of C. hepatica in porcupines. PMID- 11021438 TI - Pulmonary lymphomatoid granulomatosis in a cat. AB - Pulmonary lymphomatoid granulomatosis was diagnosed in a 9-year-old castrated male domestic shorthair cat with a history of coughing, lethargy, and anorexia. Radiographic examination revealed multiple pulmonary opacities, consolidation of left lung lobes, and enlarged tracheobronchial lymph nodes. Cytologic examination of impression smears of abnormal pulmonary tissue revealed erythrocytes, lymphocytes, and macrophages, with scattered atypical lymphocytes and binucleate cells. Histopathologic evaluation of abnormal lung tissue revealed multiple, coalescing, densely cellular nodules composed of anaplastic and pleomorphic lymphocytes, with scattered binucleate and multinucleate cells. Marked infiltration and effacement of bronchiolar and vascular smooth muscle were present. These features are characteristic of lymphomatoid granulomatosis. To the authors' knowledge, this is the first report of pulmonary lymphomatoid granulomatosis in a cat. PMID- 11021439 TI - Cardiomyopathy in captive African hedgehogs (Atelerix albiventris). AB - From 1994 to 1999, 16 captive African hedgehogs (Atelerix albiventris), from among 42 necropsy cases, were diagnosed with cardiomyopathy. The incidence of cardiomyopathy in this study population was 38%. Fourteen of 16 hedgehogs with cardiomyopathy were males and all hedgehogs were adult (>1 year old). Nine hedgehogs exhibited 1 or more of the following clinical signs before death: heart murmur, lethargy, icterus, moist rales, anorexia, dyspnea, dehydration, and weight loss. The remaining 7 hedgehogs died without premonitory clinical signs. Gross findings were cardiomegaly (6 cases), hepatomegaly (5 cases), pulmonary edema (5 cases), pulmonary congestion (4 cases), hydrothorax (3 cases), pulmonary infarct (1 case), renal infarcts (1 case), ascites (1 case), and 5 cases showed no changes. Histologic lesions were found mainly within the left ventricular myocardium and consisted primarily of myodegeneration, myonecrosis, atrophy, hypertrophy, and disarray of myofibers. All hedgehogs with cardiomyopathy had myocardial fibrosis, myocardial edema, or both. Other common histopathologic findings were acute and chronic passive congestion of the lungs, acute passive congestion of the liver, renal tubular necrosis, vascular thrombosis, splenic extramedullary hematopoiesis, and hepatic lipidosis. This is the first report of cardiomyopathy in African hedgehogs. PMID- 11021440 TI - Ossifying fibroma in a llama. AB - A 4.5-year-old llama was admitted for evaluation of a firm mass rostral and ventral to the medial canthus of the left eye. Mucopurulent nasal discharge and absence of airflow through the left nostril were noted. Radiographs of the skull revealed a sharply demarcated soft tissue mass with faint mineralization. Endoscopy of the nasal passages revealed a mucosa-covered mass originating in the area of the second premolar, extending to the edge of the soft palate, and obstructing the airway. Examination of the oral cavity revealed a missing second molar and a mass protruding 2-cm from the empty alveolus. An ossifying fibroma, a previously unreported tumor in llamas, was diagnosed at postmortem examination. PMID- 11021441 TI - Proliferation of maxillary and mandibular periodontal squamous cells in mink fed 3,3',4,4',5-pentachlorobiphenyl (PCB 126). AB - This report characterizes squamous cell proliferation in young farm mink (Mustela vison) fed a diet supplemented with 0.024 ppm 3,3',4,4',5-pentachlorobiphenyl (polychlorinated biphenyl [PCB] congener 126). One to 2 months of dietary exposure to PCB 126 resulted in gross lesions of the upper and lower jaws consisting of mandibular and maxillary nodular proliferation of the gingiva and loose teeth. The maxilla and mandible of the PCB-treated mink were markedly porous because of loss of alveolar bone. Histologically, this osteoporosis was caused by proliferation of squamous cells that formed infiltrating cords. This report clearly documents the fact that the environmental contaminant PCB 126 can cause osteoinvasive squamous proliferation in young mink, although the dose used in the present study was 7 and 36 times higher than what is typically encountered in contaminated bird eggs and fish, respectively. PMID- 11021442 TI - Tibial component fixation in porous- and hydroxyapatite-coated total knee arthroplasty: a radiostereo metric evaluation of migration and inducible displacement after 5 years. AB - A total of 51 knees (45 patients) with osteoarthrosis were stratified in a randomized study to receive a hydroxyapatite-coated Freeman-Samuelson (FS HA) or a porous-coated Miller-Galante II (MG II) uncemented total knee arthroplasty. Repeated clinical, radiographic, and radiostereometric analysis (RSA) evaluations of the tibial components were done for 5 years. The clinical outcome was equal at the 5-year follow-up (mean Hospital for Special Surgery score, 93). Standard radiographs displayed more zones around the tibial stem in the MG II group. RSA revealed that the FS HA components migrated less (smaller maximum total point motion and maximum subsidence) after 5 years and showed less inducible displacements at the 1-year follow-up. The stability of the implants obtained is equal to or better than cemented implants after 5 years. Key words: knee prosthesis, cementless, hydroxyapatite, radiostereometry, inducible displacement. PMID- 11021443 TI - The outcome of revision hip arthroplasty in patients older than age 80 years: complications and social outcome of different risk groups. AB - Between December 1995 and June 1996, we reviewed 53 consecutive patients who were at least 80 years old and had undergone a revision of a hip prosthesis between June 1988 and June 1995. Three patients (6%) had died after the operation in the hospital. Thirty-five patients were still alive with a mean follow-up of 4.0 years (range, 1.0-7.3 years). At follow-up, 15 patients had died, with a mean survival period of 25 months. Thirteen of the 15 (89%) belonged to American Society of Anesthesiologists' (ASA) group 3. Of 50 patients, 40 (80%) returned to their original social environment. Admission to a home for elderly people or to a nursing home was unrelated to the procedure in 9 of 10 cases. Complications occurred in 27 patients (51%). Major complications occurred only in high-risk patients, classified as ASA 3 according to the physical status classification of the ASA. PMID- 11021444 TI - The press-fit condylar total knee system: 8- to 10-year results with a posterior cruciate-retaining design. AB - A total of 108 consecutive Press-Fit Condylar total knee arthroplasties were performed in 94 patients. All patients had implantation with a cemented posterior cruciate-retaining design, which included resurfacing of the patella. Mean age at surgery was 70 years (range, 35-87 years). Patients were followed for a mean of 9 years (range, 8-10 years) with follow-up for all surviving patients. The average postoperative functional knee score was 96 points. Nonprogressive radiolucent lines were present in 59%. One patellar component was radiographically loose. Five knees underwent revision procedures, none for aseptic loosening. Survivorship was 93.4% at 9 years with revision for any reason as the endpoint and 98.7% with aseptic loosening as the endpoint. This knee arthroplasty shows excellent results at 8 to 10 years with no patients lost to follow-up. PMID- 11021445 TI - An in vivo determination of total hip arthroplasty pistoning during activity. AB - The purpose of this study was to determine to what extent hip joint separation occurs during normal gait on a treadmill and an abduction/adduction leg-lift maneuver in patients who have undergone total hip arthroplasty (THA). Eight patients who had a total of 10 successful unconstrained THAs (Harris Hip Scores >90) performed successive gait motions on an electronically powered treadmill and an abduction/adduction leg lift while under fluoroscopy. The fluoroscopic video images were analyzed using a 3-dimensional model-fitting technique that converts 2-dimensional fluoroscopic images into 3-dimensional real-time images. Hip joint separation was determined to be present if the amount of separation was >0.75 mm, the calculated linear error. During both activities, all 10 THAs experienced femoral head/acetabular component separation. For gait, the maximum amount of separation was 2.8 mm, while the minimum amount was 0.8 mm (average, 1.2 mm). For abduction/adduction leg lift, the maximum amount of separation was 3.0 mm, while the minimal amount was 1.7 mm (average, 2.4 mm). It appears that the femoral head separates from the acetabular component but remains in contact with the superior most tip of the component. Potential detrimental effects resulting from hip joint separation include premature polyethylene wear and component loosening. Wear may be enhanced because of the creation of multidirectional wear vectors or excessive loads resulting from eccentric femoral head pivoting. These data may be valuable in hip simulation studies to better duplicate wear patterns observed in retrieval analysis. PMID- 11021446 TI - Issues relating to long-term follow-up in hip arthroplasty surgery: a review of 598 cases at 7 years comparing 2 prostheses using revision rates, survival analysis, and patient-based measures. AB - We reviewed 598 cemented Charnley and Hi-nek total hip arthroplasties at 7 years. Data were obtained from general practitioners, hospital medical notes, microfilm, and patient questionnaires. Outcome measures were revision rates, survival analysis, 12-item Oxford Hip Score, and satisfaction ratings. There were 471 Charnley (79%) and 127 Hi-nek (21%) total hip arthroplasties; 139 deaths (23%) occurred, and 5 (<1%) were lost to follow-up. Characteristics of the Charnley and Hi-nek patient groups were similar, with more information missing for Charnley cases. Revision rates were Charnley, 37 (8%), and Hi-nek, 6 (5%) (not significant). Survival analysis revealed no difference between the 2 groups (P = .23). The patients' median Oxford Hip Score was low/good (19), slightly worse for the Hi-nek group (not significant). Taking all evidence together, neither implant was outperforming the other at 7 years. PMID- 11021447 TI - Early failure of revision total hip arthroplasty with cemented precoated femoral components: comparison with uncemented components at 2 to 8 years. AB - This is a retrospective analysis of 2 methods of fixation of the femoral component in 86 consecutive revision arthroplasties, for which all clinical and radiographic data were recorded prospectively. There were 56 cemented revisions using precoated femoral components followed for 2 to 8 years (mean, 4 years) and 30 uncemented, proximally porous-coated femoral revisions followed for 2 to 6 years (mean, 4 years). Of the 56 cemented hips, 31 (55%) had a good or excellent clinical result. Rerevision has been performed in 10 hips, and revision of 3 loose femoral components is pending (23%). Radiographic review of 56 femoral components showed that 16 (29%) had probable or definite loosening. Of 30 cementless hips, 24 (80%) had a good or excellent clinical result. There was radiographic bone ingrowth in 22 of 30 hips (73%). Seven hips (23%) had nonprogressive subsidence, and 3 hips (10%) had progressive subsidence or loosening. Rerevision has been performed in only 2 hips (7%). The high rate of loosening (29%) and rerevision (23%) at a mean follow-up of only 4 years suggests that a precoated femoral component may place increased stress at the already damaged bone-cement interface. PMID- 11021448 TI - The effect of diaphyseal biologic fixation on clinical results and fixation of the APR-II stem. AB - This study evaluated the effect of a grit-blasted diaphyseal surface on noncemented fixation of the Anatomic Porous Replacement II stem (APR-II, Sulzer Medica Orthopaedics, Austin, TX) for improvement of clinical results and fixation. A total of 107 consecutive total hip arthroplasties with the APR-II stem, which has proximal porous coating and a diaphyseal grit-blasted surface, were performed without cement, and 99 were studied at average 4 years. Of these hips, 37 had hydroxyapatite sprayed onto the proximal porous coating, but because there were no statistical differences for performance in any category, all stems were considered as 1 group. Clinical results were measured by the Harris hip score. Radiographic measurements of fixation, osteolysis, and bone remodeling were studied by reference to Gruen zones. Of hips, 99% had a good or excellent result by Harris hip score, with an average pain score of 42.3. Of hips, 98% had >40 points, with no patient reporting thigh pain after 3 years. There were no radiolucent lines in 94% of stems, and 100% had proximal bone ingrowth fixation. Distal cortical hypertrophy associated with tip fixation occurred in 49%, whereas proximal stress shielding was present in 43% of hips. Comparison of these clinical and radiographic results with our previous experience of bone ingrowth implants (smooth stem) suggests that bone ingrowth proximally with ongrowth in the diaphysis of the femoral stem provides better clinical and radiographic results. PMID- 11021449 TI - Characteristics and natural history of transient postoperative pseudosubluxation after total hip arthroplasty. AB - A series of 13 total hip arthroplasties (THAs) in which immediate postoperative radiographs revealed axial subluxation were reviewed, and the timing, incidence, and subsequent natural history of this phenomenon were determined. Each patient had received perioperative epidural anesthesia and was placed in an abduction splint before transfer to the postoperative care unit. All initial radiographs were taken while the patient was still under the effect of the epidural anesthetic agent (average, 54 minutes; range, 37-80 minutes) postoperatively. In each case, radiographs taken after anesthetic recovery and without limb manipulation revealed reduced components. The balance of the postoperative care and rehabilitation was uncomplicated; patients were followed at least 36 months postoperatively, and in only 1 patient did a subsequent hip dislocation occur. Over a 6-month period, the postoperative films of all THAs were studied, and postoperative subluxation was identified in about 3% of the cases. Postoperative subluxation is likely to be noted when the radiograph is obtained within 1 hour of transfer to the postoperative care unit. It does not appear to have a dramatic effect on the rate of postoperative dislocation, however. This radiographic finding probably is due to the continued effects of regional anesthesia, and reduction occurs as muscle tone returns. If transient subluxation is identified, no change in postoperative care is necessary. PMID- 11021450 TI - The effect of the preoperative bone quality on the fixation of the tibial component in total knee arthroplasty. AB - The relationship between the preoperative level of bone mineral density (BMD) in the proximal tibia and the migration of the tibial component 2 years after total knee arthroplasty was investigated in 28 knees with osteoarthrosis (10 men, 18 women; mean age, 71). Sixteen components were inserted uncemented and 12 were cemented. Mean average BMD measured 10 mm below the joint level was 0.81 g/cm2 (range, 0.15-1.33 g/cm2) and was not influenced by gender, age, weight, or preoperative alignment. Local BMD measured in the medial and lateral condyles was influenced by the preoperative alignment. In knees with uncemented fixation, most of the tibial component migration (ie, subsidence and lift-off) occurred within the first months, and thereafter the implants seemed to stabilize. In the uncemented implants, there was a significant relationship between average BMD and migration (regression analysis with curve-fit estimation). The least migration was seen when average BMD was 0.6 to 1.0 g/cm2. Beyond this range, increased subsidence and lift-off was seen. There was no relationship between BMD and the change in maximum migration between 1 and 2 years postoperatively, however. In knees with cemented fixation, subsidence was initially small but continuously increasing. There were no relationships between BMD and subsidence, lift-off, and maximum migration, indicating that bone-cement can compensate for variations in bone quality, at least in the early period after operation. PMID- 11021451 TI - A functional analysis of massive knee replacement after extra-articular resections of primary bone tumors. AB - We performed a functional analysis of massive knee replacements after extra articular resection of primary bone tumors. Patients did not score as highly on the Musculoskeletal Tumour Society Score and Knee Society Score (P < or = .01) as patients who had undergone intra-articular resection. Lower scores were achieved for emotional acceptance (P < or = .01), functional restriction (P < or = .05), walking distance (P < or = .05), and managing stairs (P < or = .05). These patients had a significantly greater extension lag (P < or = .01) and greater fixed flexion (P < or = .05). This functional deficit is primarily due to a compromised extensor mechanism inherent in the type of surgical resection necessary to achieve adequate removal of the tumor. PMID- 11021452 TI - The accuracy of computed tomography for determining femoral and tibial total knee arthroplasty component rotation. AB - Patellofemoral complications, instability, and tibial polyethylene wear after total knee arthroplasty (TKA) resulting from malrotation of the tibial or femoral components (or both) may be difficult to diagnose based on physical examination and standard knee radiographs. The preoperative assessment of implant rotational alignment is critical in planning treatment because the femoral or tibial component (or both) may need to be revised if malpositioned. The purpose of this study was to ascertain the accuracy of computed tomography (CT) scan for determining rotational alignment of femoral and tibial components in TKA. TKA components were inserted in human cadaver specimens at neutral and 5 degrees of external or internal rotation. For each position, the amount of rotation, determined from digital photographs, was compared with CT scan. The correlation coefficient between these two values averaged 0.87, which was significant at P < .05. The CT scan protocol described in this study can be applied clinically to patients with patellofemoral complaints to confirm or rule out the presence of component malrotation. PMID- 11021453 TI - Symptomatic osteonecrosis of the hip after orthotopic liver transplantation. AB - Patients who require long-term steroid use for immunosuppression after organ transplantation are at risk for the development of osteonecrosis. The purpose of this study was to determine the prevalence of symptomatic osteonecrosis of the hip in patients who were managed with corticosteroids for immunosuppression after orthotopic liver transplantation. We reviewed the relationship of the administration and dosage of steroids in 203 patients who underwent orthotopic liver transplantation. Overall, 29 of the 203 patients (14%) complained of hip pain, but only 3 patients (2%) developed symptomatic osteonecrosis of the hip. The osteonecrosis was diagnosed an average of 31 months after transplantation (range, 26-38 months). There was no association noted between steroid dose and the development of osteonecrosis. Routine screening for osteonecrosis of asymptomatic liver transplant patients is not necessary. PMID- 11021454 TI - Sister chromatid exchange in patients with joint prostheses. AB - The evaluation of sister chromatid exchanges (SCE) is used to establish the cytogenic damage in subjects exposed to toxic substances. The test is considered to be 1 of the most sensitive and accurate indicators of damage and responds to toxic chemicals at low doses. We evaluated the incidence of SCE in peripheral lymphocytes of patients with articular prostheses. Subjects with prostheses made of titanium-aluminium-vanadium alloys presented a significantly higher SCE number than the control population (6.3+/-2.3 vs 4.4+/-1.3; P = .0128), whereas subjects with prostheses made of chrome-cobalt alloy or mixed prostheses presented a higher SCE value than the controls but not significantly different. The presence of high-frequency cells was alarming only in 5 patients, 4 of whom had titanium alloy prostheses, whereas none belonged to the control group. The number of SCE was not affected by the presence of bone-cement used in prosthesis fixation or by the implant duration. The indication of possible cytogenic damage in patients with titanium alloy prostheses that emerged from this study should be considered carefully, even though the sample population was small. PMID- 11021455 TI - The effect of patella preparation for total knee arthroplasty on patellar strain: a comparison of resurfacing versus inset implants. AB - This study was conceived to quantify variables in surgical technique that may lead to patellar fracture after total knee arthroplasty. Anterior surface strain on load-bearing cadaveric patellae was measured before and after patellar resurfacing or inset prosthesis placement. Variables studied were i) the type of prosthesis (resurfacing vs inset), ii) the depth of reaming or osteotomy during surface preparation, and iii) the overall thickness of the polyethylene/patella composite after implantation. Comparison of measured patellar surface strain patterns for the different prosthesis styles, which were implanted at varying depths, provided statistically significant data from which the following clinically relevant conclusions can be made: i) Patellar resurfacing is superior to inset prosthesis placement when comparing postoperative patellar strain (22% vs 28% increase in strain compared with preoperative values). ii) Osteotomy for patellar resurfacing is more tolerant to error by excess cutting than is reaming for inset prosthesis placement (25% vs 42% increase in strain with a 2-mm error). iii) If the ideal depth of cut or reaming is surpassed, attempts to re-create the original patellar thickness by using a thicker prosthesis are mechanically detrimental. Key words: patella, strain, resurfacing, complications. PMID- 11021456 TI - Low-conforming all-polyethylene tibial component not inferior to metal-backed component in cemented total knee arthroplasty: prospective, randomized radiostereometric analysis study of the AGC total knee prosthesis. AB - In a prospective, randomized study of 40 patients, the quality of fixation of cemented metal-backed versus all-polyethylene tibial components of the unconstrained anatomic graduated component total knee arthroplasty design was studied during 2 years using radiostereometric analysis (RSA). The shape, as well as the articulating geometry, of the implants was identical, as was the operative technique and the postoperative regimen. In this study, no negative consequences regarding the quality of fixation using an all-polyethylene tibial component with unconstrained articulating surfaces could be identified. In all aspects, the all polyethylene tibial components displayed migration on par with, or sometimes lower than, their metal-backed counterparts. The rotations of the all polyethylene components were equally low as for the metal-backed components, and maximum lift-off was significantly lower than for the metal-backed implants. We could not identify any collapse of the bone at the medial condyle or increased subsidence at the medial part of the tibia or increased rates of radiolucent lines in the knees with all-polyethylene components. All all-polyethylene implants seemed to be stable within the resolution of RSA between 1 and 2 years, a finding known to be of positive prognostic significance regarding future aseptic loosening. PMID- 11021457 TI - Microseparation of the centers of alumina-alumina artificial hip joints during simulator testing produces clinically relevant wear rates and patterns. PMID- 11021458 TI - Underestimation of pelvic osteolysis: the value of the iliac oblique radiograph. AB - Periacetabular osteolysis is recognized as a complication of cementless total hip arthroplasty. Routine follow-up radiographs are recommended by most joint reconstructive surgeons to detect osteolytic lesions that may remain asymptomatic. These studies usually consist of an anteroposterior (AP) pelvic and lateral radiograph of the hip. Three male cadaver pelves were used to evaluate whether a routine AP pelvic radiograph was adequate to show certain osteolytic lesions. This study involved creation of progressively enlarging defects simulating an osteolytic lesion of the posterior wall. The extent of involvement of the lesion as shown by an AP pelvic radiograph and iliac oblique views was then compared. By the time the posterior column lesion could be estimated to the same extent on both the AP pelvic and iliac oblique radiographs, the lesion had become uncontained in the posterior medial direction. The evaluation of pelvic osteolysis with biplanar imaging provides a 3-dimensional understanding of the lesion. This study shows the inadequacy of a single AP pelvic radiograph in showing and localizing periprosthetic osteolytic lesions in the posterior column. PMID- 11021459 TI - Loosening of a fenestrated femoral prosthesis: a case report with histomorphologic analysis. AB - The explanted loose stem of a trabecular cementless femoral prosthesis (Holz Copf) was histologically analyzed revealing soft tissue 3 mm thick between the metal of the prosthesis and the cancellous bone. Direct contact between the implant and bone could not be found. Removal of the prosthesis was impossible by simple extraction so that a transfemoral approach was required. Prostheses with large fenestrations may be loose but at the same time ingrown with sufficient bone to require invasive procedures for their removal. PMID- 11021460 TI - Use of a magnet to retrieve a broken scalpel blade. AB - The retrieval of metallic foreign bodies, such as screws, drill bits, needles, and scalpel blades, from the surgical wound during orthopaedic surgery is often a time-consuming procedure. The extra time and dissection may result in increased morbidity. We report a case during a hip arthroplasty in which a broken surgical blade could not be located with surgical dissection even with the aid of an image intensifier. The broken blade finally was retrieved with the introduction of a magnet. PMID- 11021461 TI - Fat embolism syndrome after cementless total hip arthroplasty. AB - There are few reports in the literature of fat embolism syndrome after cementless total hip arthroplasty (THA). Most reported cases have occurred after fracture or cemented THA. We report a case of a healthy 51-year-old woman who underwent THA for osteoarthritis under spinal anesthesia. A press-fit cup and extensively porous-coated diaphyseal locking stem were used and inserted without cement. In the recovery room, the patient became hypoxemic and hypotensive and developed cortical blindness. The next day, a petechial rash was evident. Gurd's criteria for fat embolism syndrome were fulfilled. Her symptoms resolved over a 2-week period. Patients undergoing cementless THA are at risk for fat embolism syndrome, and this must be considered in the differential diagnosis for postoperative hypoxemia and neurologic deficits. PMID- 11021462 TI - Fracture of the polyethylene tibial post in posterior stabilized (Insall Burstein II) total knee arthroplasty. AB - Polyethylene wear is a well-recognized complication of total knee arthroplasty (TKA). To our knowledge, fracture of the polyethylene tibial post in a posterior stabilized TKA system has not been described as a complication or cause of failure in TKA. We report a case of a fractured polyethylene tibial post in an 85 year-old woman within 3 years of Insall Burstein II TKA, causing instability that necessitated revision surgery. PMID- 11021463 TI - Metallic instrument debris: a source of third-body wear particles? PMID- 11021464 TI - Pulmonary vein electrogram characteristics in patients with focal sources of paroxysmal atrial fibrillation. AB - INTRODUCTION: The major source of ectopic beats initiating paroxysmal atrial fibrillation (AF) is from pulmonary veins (PVs). However, the electrogram characteristics of PVs are not well defined. METHODS AND RESULTS: Group I consisted of 129 patients with paroxysmal AF. Group II consisted of 10 patients with a concealed left-sided free-wall accessory pathway. All group I patients had spontaneous AF initiated by ectopic beats, including 169 ectopic foci originating from the PVs. We analyzed PV electrograms from the 169 ectopic foci during sinus beats and ectopic beats. During AF initiation, most (70%) ectopic beats showed PV spike potential followed by atrial potential; 16% of ectopic beats showed PV fragmented potential followed by atrial potential; and 14% showed fusion potentials. The coupling interval between the sinus beat and the ectopic beat was significantly shorter in the inferior PVs than in the superior PVs (171 +/- 48 msec vs 222 +/- 63 msec, P = 0.001) and was significantly shorter in the distal foci than in the ostial foci of PVs (206 +/- 52 msec vs 230 +/- 56 msec, P = 0.01). The incidence of conduction block in the PVs during AF initiation was significantly higher in the inferior PVs than in the superior PVs (12/24 vs 37/145, P = 0.03) and was significantly higher in the distal foci than in the ostial foci of PVs (43/121 vs 6/48, P = 0.04). The maximal amplitude of PV potential was significantly larger in the left PVs than in the right PVs, and the maximal duration of PV potential was significantly longer in the superior PVs than in the inferior PVs during sinus beats in both group I and II patients. CONCLUSION: PV electrogram characteristics were different among the four PVs. Detailed mapping and careful interpretation are the most important steps in ablation of paroxysmal AF originating from PVs. PMID- 11021465 TI - Mini-maze suffices as adjunct to mitral valve surgery in patients with preoperative atrial fibrillation. AB - INTRODUCTION: After mitral valve (MV) surgery, preoperative atrial fibrillation (AF) often recurs while cardioversion therapy generally fails. Additional Cox maze surgery improves postoperative arrhythmia outcome, but the extensive nature of such an approach limits general appliance. We investigated the clinical outcome of a simplified, less extensive Cox maze procedure ("mini-maze") as adjunct to MV surgery. METHODS AND RESULTS: Thirteen patients with MV disease and preoperative AF were treated with combined surgery (group 1). Nine control patients without previous AF underwent isolated MV surgery (group 2). We retrospectively compared the results to findings in 23 patients with preoperative AF who had undergone isolated MV surgery (group 3). In group 1, mini-maze took an additional 46 minutes of perfusion time. One 75-year-old patient died of postoperative multiple organ failure. Seven patients showed spontaneously converting (within 2 months) postoperative AF. After 1 year, 82% were in sinus rhythm (SR). No sinus node dysfunction was observed. In group 2, all patients were in SR after 1 year. In group 3, only 53% were in SR after 1 year, despite serial cardioversion and antiarrhythmic drug therapy. Exercise tolerance and heart rate were comparable for groups 1 and 2. Left atrial function was present in all but one patient in group 1 and in all patients in group 2 (after MV reconstruction). CONCLUSION: Adding a relatively simple mini-maze to MV surgery improves arrhythmia outcome in patients with preoperative AF without introducing sinus node dysfunction or persistent absence of left atrial function. The results of this type of combined surgery are encouraging and deserve further attention. PMID- 11021466 TI - Recovery time dispersion measured from 87-lead body surface potential mapping as a predictor of sustained ventricular tachycardia in patients with idiopathic dilated cardiomyopathy. AB - INTRODUCTION: The clinical usefulness of QT dispersion in 12-lead ECG has been controversial in identifying subjects at risk for sustained ventricular tachycardia (VT) in patients with idiopathic dilated cardiomyopathy (DCM). We hypothesized that increasing the spatial resolution of the ECG improves the accuracy of risk stratification. The purpose of this study was to test the ability of recovery time dispersion measured from 87-lead body surface potential mapping (BSPM) to identify patients at risk for sustained VT in idiopathic DCM. METHODS AND RESULTS: We obtained 87-lead BSPM and 12-lead ECG in 33 patients with idiopathic DCM (15 patients with a history of sustained VT [VT(+) group] and 18 patients without a history of sustained VT [VT(-) group]) and in 20 normal control subjects. We measured the corrected QT dispersion and corrected recovery time dispersion from 12-lead ECG (QTc-12 dispersion and RTc-12 dispersion, respectively) and 87-lead BSPM (QTc-87 dispersion and RTc-87 dispersion, respectively). Signal-averaged ECG also was recorded in 25 patients. Neither the QTc-12 nor QTc-87 dispersion discriminated between the VT(+) and VT(-) groups patients. The VT(+) group patients had a larger but insignificant RTc-12 dispersion than the VT(-) group patients. In contrast, the RTc-87 dispersion was significantly larger in the VT(+) group patients than in the VT(-) group patients (236 +/- 39 msec vs 184 +/- 28 msec, P < 0.001). Receiver operating curve analysis indicated that the RTc-87 dispersion was as good as late potentials in predicting susceptibility to sustained VT; its sensitivity, specificity, and negative predictive value were 73%, 76%, and 76%, respectively (cutoff value 200 msec). RTc-87 dispersion >200 msec combined with positive late potentials provide high sensitivity (92%) and high negative predictive value (88%) for sustained VT. CONCLUSION: The RTc-87 dispersion is a useful tool to identify subjects at risk for sustained VT in patients with idiopathic DCM. PMID- 11021467 TI - Identification of left atrial origin of ectopic tachycardia during right atrial mapping: analysis of double potentials at the posteromedial right atrium. AB - INTRODUCTION: The high posteromedial right atrium is adjacent to the left atrium near the right superior pulmonary vein. We hypothesized that analysis of electrograms at this site could distinguish left from right atrial tachycardia. METHODS AND RESULTS: Atrial mapping was performed in 16 patients with left atrial origin ectopic tachycardia (11 patients with right superior pulmonary vein origin and 5 patients with other left atrial tachycardias). During left atrial tachycardia, earliest right atrial activation was recorded at the high posterior right atrium in 14 of 16 patients. At all of these 14 early sites, double potentials were recorded during tachycardia. The first potential was a far-field signal from left atrium as indicated by the following: (1) during sinus beats, the timing of the two potentials reversed such that the left atrial one was late; (2) ablation at the right atrial site did not decrease the amplitude of the first potential, but did decrease the amplitude of the second potential; and (3) the timing of activation at the adjacent left atrium agreed with that of the first potential. In the 11 right superior pulmonary vein tachycardias, the first potential was markedly earlier than the p wave onset, but in left atrial tachycardias with other origins it was later. In a control group of six patients with pacing to simulate right atrial tachycardia, double potentials were recorded in the posterior right atrium, but the timing of components did not reverse during sinus rhythm. CONCLUSION: For some left atrial ectopic tachycardias, particularly those originating from the right superior pulmonary vein, recognition of left versus right atrial origin can be accomplished during right atrial mapping by analysis of double potentials in the posteromedial right atrium. PMID- 11021468 TI - Mechanism for site-dependent differences in the shape of the resetting response curve in fixed barrier reentry. AB - INTRODUCTION: We investigated whether the site of stimulation within a reentrant circuit affects the resetting response curve (RRC). RRCs are used to characterize the excitable gap of reentrant circuits, including the duration of the fully excitable gap and the presence of partially excitable tissue. METHODS AND RESULTS: We reset proximal and distal to a site of interval-dependent conduction (IDC) in canine in vitro atrial tricuspid rings. Adjustable reentry allowed changes in the cycle length and direction of reentry. In nine preparations we reset 26 tachycardias. In the 16 tachycardias with one site of IDC, RRCs were significantly different when stimulating distal and proximal to a site of interval-dependent conduction. For the distal curves, the duration of the flat portion was 42 +/- 26 msec greater (P < 0.001), the slope of the increasing portion was 0.20 +/- 0.17 less (P < 0.02), and the increase in the return cycle was 14 +/- 9 msec less (P < 0.001). These differences resulted from early activation of the site of IDC by the antidromic premature impulse when stimulating from distal sites. As a result, the coupling interval of the orthodromic impulse at the site of IDC was the same or greater than at the stimulation site. In 10 tachycardias with multiple sites of IDC, significant differences in the resetting responses did not occur even when the antidromic impulse penetrated one site of IDC. CONCLUSION: In a fixed anatomic barrier reentrant circuit with one site of IDC, resetting distal to this site misrepresents the properties of the excitable gap of the entire circuit. PMID- 11021469 TI - Predictive factors of ventricular fibrillation triggered by pause-dependent torsades de pointes associated with acquired long QT interval: role of QT dispersion and left ventricular function. AB - INTRODUCTION: Death due to acquired torsades de pointes usually is caused by ventricular fibrillation (VF), but the contributing factors to VF triggered by pause-dependent torsades de pointes are not understood. METHODS AND RESULTS: We evaluated 91 patients who fulfilled four criteria: (1) pause-dependent torsades de pointes; (2) prolonged QT interval and/or corrected QT (QTc) (>0.44 sec); (3) long-short initiation sequence; and (4) conditions known to induce pause dependent torsades de pointes. There were 38 patients with a documented VF (group I) and 53 without VF (group II). Absolute and relative dispersions of QT and QTc were calculated based on the 12-lead standard ECG. Group I differed from group II with regard to myocardial infarction history (32% vs 13%; P = 0.035), left ventricular ejection fraction (44% +/- 14% vs 65% +/- 9%; P < 0.0001), presence of structural heart disease (100% vs 20.8%; P < 0.0001), QT mean (591 +/- 73 msec vs 514 +/- 78 msec; P < 0.0001), QTc mean (563 +/- 76 msec vs 508 +/- 90 msec; P = 0.002), absolute QT dispersion (166 +/- 56 msec vs 84 +/- 49 msec; P < 0.0001), relative QT dispersion (9.9% +/- 3.5% vs 6.3% +/- 3.2%; P < 0.0001), absolute QTc dispersion (158 +/- 57 msec vs 81 +/- 44 msec; P < 0.0001), and relative QTc dispersion (9.9% +/- 3.6% vs 6.2% +/- 3%; P < 0.0001). Multiple regression analysis showed that ejection fraction (P = 0.0001), presence of structural heart disease (P < 0.0001), and relative QTc dispersion (P = 0.038) were the only independent predictors of VF. CONCLUSION: Left ventricular function, presence of structural heart disease, and QTc relative dispersion should be evaluated carefully in patients with conditions susceptible to inducing torsades de pointes. PMID- 11021470 TI - Reversal of repolarization gradient does not reverse the chirality of shock induced reentry in the rabbit heart. AB - INTRODUCTION: Two hypotheses have been proposed to explain the mechanisms of vulnerability and related failure of defibrillation therapy: the cross-field induced critical point hypothesis and the virtual electrode-induced phase singularity hypothesis. These two hypotheses predict the opposite effect of preshock repolarization on the chirality (direction of rotation) of shock-induced reentry. The former suggests its reversal upon reversal of repolarization, whereas the latter suggests its preservation. The aim of this study was to determine, by reversing the repolarization sequence, which of the mechanisms is responsible for internal shock-induced arrhythmia in the Langendorff-perfused rabbit heart. METHODS AND RESULTS: We used high-resolution optical mapping to assess the chirality of postshock reentry in 11 hearts. Hearts were paced at a coupling interval of 300 msec at various sites around the field of view (13.5 x 13.5 to 16.5 x 16.5 mm). Cathodal monophasic implantable cardioverter defibrillator shocks (-100 V, 8 msec) were applied during the T wave from a 10-mm coil electrode placed into the right ventricular cavity. We used 3.5 +/- 0.8 different pacing sites per heart. Change in direction of repolarization did not result in change of chirality. Chirality was constant in all 11 hearts despite the complete reversal of activation and repolarization patterns. However, the position of resulting vortices depended on transmembrane polarization gradient inverted delta Vm and amplitude of negative polarization Vm (deexcitation). Stronger gradients and deexcitation produced earlier epicardial break excitation (P = 0.04 and P < 0.0001, respectively). CONCLUSION: Virtual electrode-induced phase singularity mechanism underlies internal shock-induced arrhythmia in this model. PMID- 11021471 TI - Defibrillation mechanisms: the parable of the blind men and the elephant. PMID- 11021472 TI - Prediction of defibrillation outcome by epicardial activation patterns following shocks near the defibrillation threshold. AB - INTRODUCTION: Ventricular defibrillation is probabilistic and shock strength dependent. We investigated the relationship between defibrillation outcome and postshock activation patterns for shocks of the same strength (approximately 50% probability of success for defibrillation [ED50] to yield an equal number of successful and failed shocks). METHODS AND RESULTS: In five pigs, 10 shocks of approximately ED50 strength (right ventricle-superior vena cava, biphasic, 6/4 msec) were delivered after 10 seconds of ventricular fibrillation (VF). Epicardial activation sequences following shocks were mapped with a 504-electrode shock and analyzed by animating dV/dt of the electrograms. Intercycle interval (ICI, time between the onset of successive postshock cycles), wavefront conduction time (WCT, time between the earliest and latest activation of a cycle), and overlapping index (WCT of cycle[n]/ICI of cycle[n+1]) were determined for the first five postshock cycles. An overlapping index >1 indicates overlap between successive cycles. Of 50 defibrillation attempts, 25 were successes. There was no difference between successful and failed episodes for both ICI (68 +/- 9 msec vs 62 +/- 10 msec) and WCT (97 +/- 24 msec vs 100 +/- 14 msec) of cycle 1. However, starting at cycle 2, the ICI was longer, and the WCT was shorter for successful than failed episodes (P < 0.01). Overlapping cycles (index > 1) were found during the transition from cycles 2 through 5 in all failed (index >1) and in no successful episodes. CONCLUSIONS: (1) Defibrillation outcome cannot be determined during the first postshock cycle. (2) At least three rapid successive cycles with overlap of cycles 2 and 3 are present in all failed and in no successful episodes. (3) The overlapping index is a marker to predict defibrillation outcome. PMID- 11021473 TI - Pacing following shocks stronger than the defibrillation threshold: impact on defibrillation outcome. AB - INTRODUCTION: A recent study of shocks near defibrillation threshold (DFT) strength demonstrated that at least three rapid cycles always occur after failed shocks but not after successful shocks, suggesting that the number and rapidity of postshock cycles are important in determining defibrillation success. To test this hypothesis, rapid pacing was performed following a shock stronger than the DFT that by itself did not induce rapid cycles and ventricular fibrillation (VF). METHODS AND RESULTS: Epicardial activation was mapped in six pigs using a 504 electrode sock. The DFT was determined by an up/down protocol with S1 shocks (right ventricle-superior vena cava, biphasic). Ten shocks that were 100 to 200 V above the DFT (aDFT) were delivered after 10 seconds of VF to confirm they always defibrillated. Then, S2, S3, etc., pacing at 5 to 10 times diastolic threshold was performed from the left ventricular apex after aDFT shocks during VF. First, the postshock interval after aDFT shocks was scanned with an S2 stimulus to find the shortest S1-S2 coupling interval (CI) that captured. This was repeated for S3, S4, etc., until VF was induced. To induce VF after aDFT shocks, three pacing stimuli (S2, S3, S4) with progressively shorter CIs were always required; S2 or S2,S3 never induced VF. For the S2-S4 cycles, the intercycle interval was shorter (P < 0.01), and the wavefront conduction time was longer (P < 0.01) for episodes in which VF was induced (n = 57) than for episodes in which it was not (n = 60). Following the S4 cycle that induced VF, two types of spontaneous activation patterns appeared: focal (88%) and reentrant (12%). CONCLUSION: VF induction after aDFT shocks always required at least three rapid successive paced-induced cycles. Thus, the number and rapidity of the first several postshock cycles rather than just the first postshock cycle may be determining factors for defibrillation outcome. PMID- 11021474 TI - Spatial distribution and frequency dependence of arrhythmogenic vagal effects in canine atria. AB - INTRODUCTION: Prior studies in isolated canine atria demonstrated that acetylcholine-induced reentrant atrial fibrillation (AF) was triggered by multifocal activity in the area of normal impulse origin (sinus node-crista terminalis). The aim of this study was to investigate the activation sequence in AF induced by vagal stimulation (VS) in intact dog hearts. METHODS AND RESULTS: VS (10 to 50 Hz, 1 msec, 15 V, 5-sec trains) induced single or multiple atrial premature depolarizations (APDs), and/or AF in 8 of 10 open chest dogs. Occurrence of APDs and AF increased with increasing VS intensity. Epicardial mapping (254 unipolar electrodes) of both atria showed that APDs as a rule emerged from ectopic sites, often from the right atrial appendage. Activation mapping of the first 10 cycles of AF showed that only a small number (<3 to 4) of unstable reentrant circuits were possible at the same moment. Moreover, most sustained VS-induced AFs were accounted for by a single leading stable reentrant circuit that activated the remainder of the atria. CONCLUSION: (1) Occurrence of vagally induced APDs and AF increases with increasing frequency of VS. (2) VS induced focal ectopic APDs are widely distributed over the atria. (3) A single APD can be sufficient for initiation of reentrant AF. (4) Despite its high rate of sustained AF, it may be maintained by single stable reentrant circuit. (5) The atrial septum can play an important role in both the initiation and the maintenance of VS-induced AF. PMID- 11021475 TI - Donor-to-recipient decremental conduction of atrial fibrillation following orthotopic heart transplantation: insights into the mechanism of atrioatrial conduction. AB - We describe the unique case of a heart transplant patient with type I atrial fibrillation that arose in the donor atrium during a late acute rejection episode and conducted to the recipient atrium with second-degree type I local block. After internal cardioversion, programmed stimulation showed bidirectional decremental conduction across the suture line with nearly equal atrioatrial interval, whereas the recipient atrium showed progressively delayed intra-atrial conduction. These findings strongly suggest that the mechanism of atrioatrial conduction may be electrical propagation along viable myocardium bridging the surgical scar and that the electrophysiologic characteristics of the recipient atrium are involved in decremental conduction across the suture line. PMID- 11021476 TI - Hypokalemia-induced long QT syndrome with an underlying novel missense mutation in S4-S5 linker of KCNQ1. AB - Congenital long QT syndrome (LQTS) is caused by mutations in at least five genes coding for cardiac potassium or sodium channels that regulate the duration of ventricular action potentials. Acquired LQTS often is associated with drugs or metabolic abnormalities. A 47-year-old woman who presented with marked QT prolongation (QTc = 620 msec(1/2)) and repeated episodes of torsades de pointes associated with hypokalemia (2.6 mEq/L) was screened for mutations in LQTS genes using polymerase chain reaction/single-strand conformation polymorphism (PCR/SSCP). We identified a novel missense mutation in the intracellular linker of S4-S5 domains of KCNQ1, resulting in an amino acid substitution of cysteine for arginine at position 259 (R259C). Whole cell, patch clamp experiments were conducted on COS7 cells transfected with wild-type and/or R259C KCNQ1 with or without KCNE1. Functional analyses of the mutant KCNQ1 subunit on COS7 cells revealed its functional channels in the homozygous state, producing a significantly smaller current than the KCNQ1 channels and a less severe dominant negative effect on I(Ks). The novel KCNQ1 mutation R259C is the molecular basis for I(Ks) dysfunction underlying an apparently sporadic case of hypokalemia induced LQTS, consistent with a mild mutation likely to disclose the clinical manifestation of LQTS in a context of severe hypokalemia. Our findings suggest that gene carriers with such mild mutations might not be so rare as commonly expected in patients with acquired LQTS, and stress the importance of mutational analysis for detecting either "silent" forms of congenital LQTS or de novo mutations. PMID- 11021477 TI - Phenotype (ECG)-genotype considerations in long QT syndrome and Brugada syndrome. PMID- 11021478 TI - Preexcited tachycardia: what is the tachycardia mechanism? PMID- 11021479 TI - EP images: from cell to bedside. Spot welding the trigger in focal atrial fibrillation ablation. PMID- 11021480 TI - A positive oesophageal margin in stomach cancer. PMID- 11021481 TI - Extrafascial excision for rectal cancer. PMID- 11021482 TI - Significance of a positive oesophageal margin in stomach cancer. AB - BACKGROUND: A positive oesophageal margin is frequently encountered in total and proximal gastrectomies. It is controversial as to whether a positive oesophageal margin in gastrectomy predisposes to anastomotic dehiscence and loco-regional recurrence. Its independent impact on survival has not been fully addressed. METHODS: A retrospective review of 137 total and proximal gastrectomies for adenocarcinoma was undertaken. Independent prognostic factors were identified in a bivariate and Cox proportional hazards regression model. RESULTS: The prevalence of positive oesophageal margin was 18.2%. A positive oesophageal margin was not associated with increased anastomotic leak (8.0 vs 10.7%; P = 0.51), operative morbidity (32.0 vs 27.7%; P = 0.85) or 30-day mortality (8.0 vs 5.4%; P = 0.48) rates when compared with a negative margin. In addition, it did not predispose to anastomotic (13.0 vs 10.4%; P = 0.47) or regional (22.7 vs 24.5%; P = 0.51) recurrences. In the multivariate analysis the TNM stage and status of the oesophageal margin were the only independent prognostic factors for survival. CONCLUSIONS: A positive oesophageal margin is an independent poor prognostic factor for long-term survival in stomach cancer. All efforts should therefore be made to clear the oesophageal margin in total and proximal gastrectomies. PMID- 11021483 TI - Results of extrafascial excision and conventional surgery for rectal cancer at Auckland Hospital. AB - BACKGROUND: Recent studies have suggested that local recurrence rates following rectal cancer surgery are reduced if the mesorectum is removed intact within its fascia propria. The present study aims to compare the outcomes of conventional surgery for rectal cancer and surgery in which the rectum and mesorectum are removed by the technique of extrafascial excision (EFE). METHODS: All patients undergoing surgery for rectal cancer at Auckland Hospital from 1980 to 1996 were identified. Demographic, tumour, operation, outcome, survival and follow-up data were obtained from patient charts, New Zealand (NZ) Death Registry, death certificates and the NZ Electoral Roll. Complication rates, recurrence rates, overall and cancer-free survival and treatment costs were calculated for each group. RESULTS: A total of 262 patients had curative surgery (138 had conventional surgery, 124 had EFE). The groups were similar with respect to age, sex, operation performed and Dukes' stage. There was no difference in complication rates between the groups. Mean follow-up was 7 years in survivors. Twenty-nine conventional-surgery (21%) and eight EFE (6%) patients developed local pelvic recurrence. The 5-year actuarial local recurrence rates were 30% and 10%, respectively (P = 0.0006). The 5-year overall survival was 54% for conventional surgery and 60% for EFE (P = 0.23). The 5-year cancer-free survival was 63% for conventional surgery and 74% for EFE (P = 0.02). Average initial costs were NZ$15,717 and NZ$15,158 for conventional surgery and EFE, respectively. The average cost of local recurrence was an additional NZ$10,471. CONCLUSIONS: The present study adds further support to the growing evidence that excision of the mesorectum within an intact fascial envelope reduces local recurrence rates after surgery for rectal cancer. There appears to be an associated improvement in cancer-free survival. Complication rates and cost were not increased in the patients having EFE. PMID- 11021484 TI - Consultative Committee on Road Traffic Fatalities: trauma audit methodology. AB - BACKGROUND: Since 1992 the Consultative Committee on Road Traffic Fatalities in Victoria has identified deficiencies and errors in the management of 559 road traffic fatalities in which the patients were alive on arrival of ambulance services. The Committee also assessed the preventability of deaths. Reproducibility of results using its methodology has been shown to be statistically significant. The Committee's findings and recommendations, the latter made in association with the learned Colleges and specialist Societies, led to the establishment of a Ministerial Taskforce on Trauma and Emergency Services. As a consequence, in 2000, a new trauma care system will be implemented in Victoria. This paper presents a case example demonstrating the Committee's methodology. METHODS: The Committee has two 12 member multidisciplinary evaluative panels. A retrospective evaluation was made of the complete ambulance, hospital and autopsy records of eligible fatalities. The clinical and pathological findings were analysed using a comprehensive data proforma, a narrative summary and the complete records. Resulting multidisciplinary discussion problems were identified and the potential preventability of death was assessed. RESULTS: In the present case example the Committee identified 16 management deficiencies of which 11 were assessed as having contributed to the patient's death; the death, however, was judged to be non-preventable. CONCLUSION: The presentation of this example demonstrating the Committee's methodology may be of assistance to hospital medical staff undertaking their own major trauma audit. PMID- 11021485 TI - Pancreatic cancer and diabetes: is there a relationship? A case-controlled study. AB - BACKGROUND: There is controversy about whether diabetes mellitus is a risk factor for pancreatic cancer or an epiphenomenon of the cancer. The present study aims to determine if long-term diabetes is a risk factor for pancreatic cancer. METHODS: The study undertook to determine the prevalence of diabetes among three matched (age/gender) patient groups (pancreatic cancer (PaC), colorectal cancer (CRC), and fracture neck of femur (NOF)) at the date of diagnosis of cancer or fracture as well as 1 and 5 years prior to this. A retrospective review of the medical records of the three groups of patients was undertaken. Patients identified with PaC in the period July 1994 to February 1998 were age (+/- 5 years)- and gender-matched to patients identified in the same time period with NOF and with CRC. The data were then analysed using McNemar's test for discordant pairs. RESULTS: Over a 44-month period 116 patients with PaC were identified of which 24% had diabetes at the time of diagnosis of their malignancy (NOF, 8%; CRC, 9.5%). There was a statistically significant difference (PaC and NOF, P < 0.01; PaC and CRC, P < 0.01). For a duration of diabetes of > 5 years the prevalence of diabetes fell to 7.8% in the PaC group, to 6% in the NOF group and to 6.9% in the CRC group, with no significant difference between the groups. CONCLUSION: There is no increase in the prevalence of long-standing diabetes mellitus in patients with PaC compared to age- and gender-matched controls with NOF and CRC. The relationship of PaC and diabetes may be an epiphenomenon, rather than diabetes being a risk factor for pancreatic malignancy. PMID- 11021486 TI - Absolute risk of breast cancer for Australian women with a family history. AB - BACKGROUND: The purpose of the present paper was to estimate the absolute risk of breast cancer over the remainder of a lifetime in Australian women with different categories of family history. METHODS: Age-specific breast cancer incidence rates were adjusted for screening effects, and rates in those with no family history were estimated using the attributable fraction (AF). Relative risks from a published meta-analysis were applied to obtain incidence rates for different categories of family history, and age-specific incidence was converted to cumulative risk of breast cancer. The risk estimates were based upon Australian population statistics and published relative risks. Breast cancer incidence was from New South Wales women for 1996. The AF was calculated using prevalence of a family history of breast cancer from data on Queensland women. The cumulative absolute risk of breast cancer was calculated from decade and mid-decade ages to age 79 years, not adjusted for competing causes of death. RESULTS: Lifetime risk is approximately 8.6% (1 in 12) for the general population and 7.8% (1 in 13) for those without a family history. Women with one relative affected have lifetime risks of 1 in 6-8 and those with two relatives affected have lifetime risks of 1 in 4-6. The cumulative residual lifetime risk decreases with advancing age; by age 60 years all groups with only one relative affected have well above a 90% probability of not developing breast cancer to age 79 years. CONCLUSIONS: These Australian risk statistics are useful for public information and in the clinical setting. Risks given here apply to women with average breast cancer risk from other risk factors. PMID- 11021487 TI - Management of intraparotid facial nerve schwannomas. AB - BACKGROUND: The purpose of the present paper was to review the management of intraparotid facial nerve schwannoma so as to discuss its clinical presentation, evaluate the various possible diagnostic investigations, and compare the various surgical options and outcome. METHODS: Case series was undertaken of five (1.3%) patients with facial nerve schwannoma out of 400 consecutive parotidectomies at Singapore General Hospital. RESULTS: There were three men and two women with an age range of 29-65 years. Three patients presented with painless parotid lumps while two had painful parotid swellings. None had facial nerve paresis. Only one patient had preoperative diagnosis suspicious of schwannoma by fine-needle aspiration cytology (FNAC). Diagnoses were made intraoperatively. Four patients had excision with cable grafting of the nerve defect. achieving facial nerve grade II-IV (House-Brackmann scale). One patient who underwent enucleation of tumour with nerve preservation achieved grade II. CONCLUSIONS: Preoperative diagnosis is difficult but it is important for discussion of the extent and options of surgery. Fine-needle aspiration cytology holds promise in making a preoperative diagnosis. Enucleation with nerve preservation where possible seems to offer better facial function whereas nerve excision with cable graft can give satisfactory results. PMID- 11021488 TI - Anaesthetic simulators: training for the broader health-care profession. AB - BACKGROUND: The use of high-fidelity patient simulators for training health-care professionals has increased rapidly in recent years. Approximately 150 simulation training centres operate internationally. Australasia has acquired four centres since 1997. A large component of simulator-based training is experiential. METHODS: Participants manage clinical scenarios on lifelike computer-controlled mannikins within realistic clinical environments. Afterwards they actively reflect upon the experience, an exercise that is facilitated by observation of a video replay of the event. RESULTS: This approach to training promotes a consideration of broader issues which can influence clinical practice and patient outcomes. This has particular relevance to emergencies. Here, events that are by nature infrequent and unscheduled can be addressed in a controlled fashion, in an environment that is supportive and separated from actual patients. CONCLUSIONS: A broad range of skills can be addressed with this resource. Of key importance are situational management and team effectiveness skills. Deficiencies with respect to these 'non-clinical' skills are being increasingly identified for their contribution to preventable adverse events within the health-care environment. Multidisciplinary operation-room team training has the potential to address these issues as they relate to the perioperative environment. PMID- 11021489 TI - Effects of CO2 insufflation and laparotomy on wound healing in mice. AB - BACKGROUND: The aim of the present study was to evaluate the effects of laparotomy and CO2 insufflation on wound healing in a murine incisional wound healing model. METHODS: Seventy-two male Swiss Albino mice were randomly allocated into three groups of control, laparotomy and CO2 insufflation. A transverse skin incision of 15 mm was made in the dorsum of each mouse, and four interrupted mattress sutures with 4.0 polypropylene thread were laid for wound closure. A median laparotomy was performed in the laparotomy group. CO2 insufflation was performed with an intra-abdominal pressure of 9 mmHg. The retained gas was evacuated from the abdominal cavity at the end of a 60-min period. Mice were killed on the 3rd, 7th and 15th postoperative days. The wound tensile strength and 5-hydroxyproline concentration in the wound tissue were measured. RESULTS: Tensile strength of the incised skin increased as the post incision period progressed. There was no significant difference between the tensile strengths of the incised skin of control, laparotomy and CO2 insufflation groups throughout the observation period. The skin 5-hydroxyproline concentrations of all groups were not significantly different at the 3rd postoperative day. But laparotomy and CO2 insufflation groups had lower 5 hydroxyproline concentrations at the 7th and 15th postoperative days, when compared to controls (P < 0.02 for 7th and 15th days). CONCLUSION: CO2 insufflation and laparotomy reduce the 5-hydroxyproline concentration of the wound, suggesting a diminished wound healing capacity. PMID- 11021490 TI - A prospective randomized trial of day-stay only versus overnight-stay laparoscopic cholecystectomy: comment. PMID- 11021491 TI - Minimally invasive surgery for primary hyperparathyroidism: a systematic review: comment. PMID- 11021492 TI - Rectal Dieulafoy's lesion: cause of massive lower gastrointestinal tract haemorrhage. PMID- 11021493 TI - Postoperative incisional fistula involving a Meckel's diverticulum. PMID- 11021494 TI - Successful endoscopic treatment of enterocutaneous fistulas by histoacryl glue. PMID- 11021495 TI - Diffuse oncocytic hyperplasia of the parotid gland. PMID- 11021496 TI - Malignant melanoma originating in the thymus. PMID- 11021497 TI - Leo Doyle, master surgeon. AB - On 3 March 1953 Leo Doyle died at the Mercy Hospital, Melbourne. The day before he died Leo Doyle had been operating at the Mercy Hospital when he took ill. Doyle's final illness was almost certainly the result of the severe aortic stenosis that had been developing over some years. His death at the relatively young age of 61 ended the career of a man described by Sir Gordon Gordon Taylor as the greatest technical surgeon that he had ever seen. In all likelihood Australian surgery will never see the likes of Doyle, a virtuoso surgeon, again. And yet to many of the surgeons who were Doyle's contemporaries and to those who followed him he remained somewhat of an enigma. Perhaps in some way the description of the great French surgeon Baron Dupuytren may also be applicable to Leo Doyle: known to all, loved by many, understood by few. By all accounts Leo Doyle's surgical repertoire knew no bounds. He operated with equal facility on the central nervous system, the head and neck, in the thorax, abdomen and pelvis and he was more than competent in gynaecology, urology and orthopaedics. In the latter part of his career he became, par excellence, a cancer surgeon. He was, arguably, Australia's first surgical oncologist. No procedure was deemed too complicated or demanding. Like some other superb technicians his judgement at times did not match his technical ability. Doyle was one of the first surgeons in Australia to perform hindquarter amputation and he helped to pioneer the operations of total gastrectomy and oesophagogastrectomy. An avid reader of the surgical literature, he possessed an enormous library which was matched by an equally large collection of surgical instruments. Unlike Devine he published relatively little. He was not a good clinical teacher, preferring to teach by example in the operating theatre. Although interested in music and the visual arts, surgery was his life. PMID- 11021498 TI - Perceptual and acoustic study of professionally trained versus untrained voices. AB - Acoustic and perceptual analyses were completed to determine the effect of vocal training on professional singers when speaking and singing. Twenty professional singers and 20 nonsingers, acting as the control, were recorded while sustaining a vowel, reading a modified Rainbow Passage, and singing "America the Beautiful." Acoustic measures included fundamental frequency, duration, percent jitter, percent shimmer, noise-to-harmonic ratio, and determination of the presence or absence of both vibrato and the singer's formant. Results indicated that, whereas certain acoustic parameters differentiated singers from nonsingers within sex, no consistently significant trends were found across males and females for either speaking or singing. The most consistent differences were the presence or absence of the singer's vibrato and formant in the singers versus the nonsingers, respectively. Perceptual analysis indicated that singers could be correctly identified with greater frequency than by chance alone from their singing, but not their speaking utterances. PMID- 11021499 TI - Study of the elderly females' voice by phonetography. AB - This study was carried out to investigate the voice characteristics of 40 healthy females with no voice disorders, ranging in age from 60 to 84 years (X = 68.2+/ 5.74 years). Measurements over all the entire phonational range were obtained by phonetography. The subjects were asked to sustain the vowel /a/ in modal register for a minimum of 5 seconds in the highest and lowest intensities after hearing the semitones C, E, G, and A, over all phonational ranges. The results indicated expansion of the low and reduction of high ends of the pitch range, decrease of the pitch numbers of the vocal range, restriction of the lowest and highest limits of the intensity, and reduction of either the maximum phonational range and the phonetogram area. The phonetography technique has shown efficacy to investigate the voice characteristics of elderly females. PMID- 11021500 TI - Effects of stimulation techniques on vocal responses: implications for assessment and treatment. AB - During voice evaluation and treatment it is customary for clinicians to elicit samples of the vowel /a/ from clients using various elicitation techniques. The purpose of this study was to compare the effects of four commonly used stimulation tasks on the laryngeal mechanism. Eleven female singing students, studying at a university music school, served as subjects for the study. The subjects phonated the vowel /a/ using 4 vocal stimulation techniques: yawn-sigh, gentle onset, focus, and the use of the voiceless fricative. Videoendoscopic and acoustic evaluations of their productions were done. Results show that, in the first 100 ms following the end of the formant transition, these techniques affected voice differently. The fundamental frequency was found to be highest in the yawn-sigh condition, whereas the maximum frequency perturbation was obtained for the voiceless fricative condition. Planned comparisons were made by comparing the data across 2 dimensions: (1) vowels elicited with voiced contexts versus those elicited with voiceless consonantal contexts and (2) vowels elicited with obstruent versus vowels elicited with nonobstruent consonantal contexts. Some changes in acoustic parameters brought about by these stimulation techniques may be explained on the basis of coarticulatory effects of the consonantal context. PMID- 11021501 TI - Respiratory and laryngeal contributions to maximum phonation duration. AB - Maximum phonation duration (MPD) is a common assessment procedure in speech language pathology. However, the specific contributions of the respiratory and phonatory components of the speech-production mechanism to this task are not typically assessed. Six women and 6 men with normal speech and voice were monitored for lung volume during a standard MPD task, and for laryngeal airway resistance (Rlaw) during a modified MPD (slow syllable-repetition) task. On average, subjects used 90% of their vital capacity (VC) for their best MPD trial. There was no systematic relation between MPD and VC for these subjects. Rlaw was strongly correlated with MPD for the men (r(s) = 0.886 for /a/; r(s) = 0.829 for /i/), but not for the women. Rlaw increased linearly as lung volume decreased (slope > 0.15) for a subset of trials (32%). This was a common pattern for four of the subjects. The clinical utility of MPD to assess breathing for speech is questioned because of the lack of association between MPD and VC, and some atypical laryngeal-valving strategies. PMID- 11021502 TI - Effects of a vocally fatiguing task and systemic hydration on phonation threshold pressure. AB - Phonation threshold pressure (PTP), effort for speaking, and vibratory closure pattern were assessed in 4 women with normal untrained voices after 2 hours of loud reading. PTP generally increased after this vocally fatiguing task at conversational pitch and 10%, 50%, and especially 80% of the pitch range. Increased systemic hydration by drinking water appeared to attenuate and/or delay the elevation of PTP for 3 subjects, at least at the highest pitch tested. Effort for speaking increased consistently throughout the loud reading task and subsequently decreased after 15 minutes of vocal silence. Upon videostroboscopic examination of the larynx, 3 subjects demonstrated spindle-shaped vibratory closure patterns on occasion after loud reading. The results provide preliminary support for increasing water consumption to reduce or delay some vocal-function changes after prolonged loud phonation in untrained speakers. PMID- 11021503 TI - Taxonomy of singers used as subjects in scientific research. AB - Criteria for determining the classification of singers in research has posed numerous problems for both researchers and their readership. Previous attempts at creating usable categories have failed because of disagreement within the professions involved in such research. The object of this paper is to explore and suggest a system in which scientific research related to singing is based on a quantifiable singing standard. Use of the systematic categorization set out in this paper will help to alleviate the questions regarding performance abilities of singers used as subjects in scientific projects and lead to more credible results. Nine singer categories have been consolidated and based on proven performance achievement. These categories range from superstar (household name) to local community and amateur singers with comprehensive subcategories under each topic for each type of singer, that is, opera, pop, jazz, etc. PMID- 11021504 TI - Voice handicap index in singers. AB - The Voice Handicap Index (VHI) was developed to assess patients' perception of the severity of their voice disorder. The purpose of this study was to determine the degree of handicap expressed by professional and recreational presenters with a voice complaint. Singers (n = 106) and nonsingers (n = 369) with voice symptoms were studied. The results of the VHI for singers indicate that singers score significantly lower (less severe) on the VHI compared to nonsingers. Singers with vocal fold nodules had a lower mean VHI than singers with vocal fold cysts or polyps. Singers who perform classical music had the lowest mean VHI of all types of singers studied. A low VHI in singers may represent a significant handicap and should not be ignored when considering the severity of a singer's voice problem. PMID- 11021505 TI - Voice-related quality of life (V-RQOL) following type I thyroplasty for unilateral vocal fold paralysis. AB - Unilateral vocal fold paralysis is a common clinical problem which frequently causes severe dysphonia. Various treatment options exist for this condition, with the type I thyroplasty being one of the more commonly performed surgical procedures for vocal rehabilitation. The Voice-Related Quality of Life (V-RQOL) Measure is a validated outcomes instrument for voice disorders. This study measured the V-RQOL of patients with unilateral vocal fold paralysis who had undergone a type I thyroplasty and compared these scores to those of patients with untreated and uncompensated unilateral vocal fold paralysis and to normals. Treated patients had significantly higher domain and overall V-RQOL scores than untreated patients, but also scored lower than normals. These differences were true across gender and age. Patients who were more distant from surgery had lower V-RQOL scores than those who had more recently been treated. It is concluded that type I thyroplasty leads to a significantly higher V-RQOL for patients with unilateral vocal fold paralysis. This study also demonstrates further the utility of patient-oriented measures of treatment outcome. PMID- 11021506 TI - Soft tissue response in the rabbit larynx following implantation of LactoSorb (PLA/PGA copolymer) prosthesis for medialization laryngoplasty. AB - This project is designed to provide initial data regarding the use of polylactic acid/polyglycolic acid (PLA/PGA) copolymer ("LactoSorb" [Walter Lorenz Corp]), an alloplastic, resorbable material, as a prosthesis in an animal model of vocal fold medialization. Fifteen New Zealand white rabbits were utilized for left medialization laryngoplasty with LactoSorb implants after undergoing left recurrent laryngeal nerve section. At 1, 3, 6, and 9 months, the rabbits were sacrificed and their larynges were evaluated both grossly and histologically for tissue response to, and resorption characteristics of the implant, tissue cellularity, maintenance of vocal fold medialization, and airway patency. Additionally, 4 rabbits were used as controls, implanted with silicone rubber medialization implants, and sacrificed at 9 months for comparison. One rabbit underwent no surgery and was likewise used as a control. Grossly, no airway obstruction was noted, and no extrusions of the implants occurred. The LactoSorb implant maintained medialization in each group of sacrificed rabbits. Histologic findings revealed a very discrete, fibrous capsule around the implant in the 1- and 3-month rabbits, and the LactoSorb was still grossly visible. At 6 months, the thin fibrous capsule partially remained; at 9 months, the capsule was no longer evident, and the implant was no longer grossly visible. Endoscopic findings at the time of sacrifice in those rabbits implanted with silicone rubber included grossly patent airways with maintenance of medialization. In the rabbits implanted with silicone rubber, the histologic findings are similar to those described elsewhere. LactoSorb, because of its intermittent resorption rate, could offer an ideal alternative to currently utilized temporary, or resorbable, materials, and as such will hopefully prove an invaluable tool in the laryngologist's treatment planning and surgical repair of the patient with a paralyzed vocal fold. PMID- 11021507 TI - "Alternative medical therapy" use among singers: prevalence and implications for the medical care of the singer. AB - Singers are extremely conscious of health problems that affect their voices and well-being and often take an active role in seeking care for these problems. They frequently seek treatment from providers or with modalities considered "alternative" to traditional medical care. A survey of singers was completed to elucidate their attitudes and practices regarding "alternative modalities" of medical care. Frequently singers will self-medicate or take advice from people not well versed in the special needs of a professional voice user. They will fail to share this information with the physician when seeking "traditional" medical care. These practices may predispose the singer to suboptimal or even dangerous care. These results are discussed, as well as the implications for the medical physician treating the singer. The possible detrimental pharmacologic effects of "natural therapies" widely used by singers are presented, with special attention to the particular concerns for the professional singer. PMID- 11021508 TI - The treatment of essential voice tremor with botulinum toxin A: a longitudinal case report. AB - The purpose of this study was to evaluate the effects of bilateral botulinum toxin injection into the thyroarytenoid (TA) muscles of a patient with essential voice tremor. Acoustic and aerodynamic data were collected weekly over a 16-week period. Flexible nasolaryngoscopy was performed prior to injection and 2, 6, 10, and 16 weeks postinjection. Perceptual analyses of the acoustic and nasolaryngoscopic data were performed. A reduction in frequency tremor and, to a lesser extent, amplitude tremor was observed during the 1-10 week period. Estimated laryngeal resistance decreased after injection and was accompanied in perceptual measures by a reduction in vocal effort, laryngeal tremor, and supraglottic hyperfunction. Essential voice tremor can be successfully attenuated with bilateral percutaneous injection of botulinum toxin A into the vocalis muscle. PMID- 11021509 TI - Functional imaging of vocal fold vibration: digital multislice high-speed kymography. AB - For the diagnosis of voice disorders, and especially for the classification of hoarseness, direct observation of vocal fold vibration is essential. Furthermore, a quantitative description of the movement of the vocal fold becomes increasingly necessary to document and compare findings as well as the progression of speech therapy. On the base of digital high-speed sequences of vocal fold vibration, multiple "functional images"-also called digital kymograms-are obtained using image- and signal-processing algorithms. Digital kymograms can serve as a powerful aid for visualization, description, and classification of vocal fold vibration and as an intermediate step for image interpretation by biomechanical modeling. This visualization technique will be discussed and compared to other techniques currently available: videokymography and videostroboscopy. The technique is applied to several clinical examples: aperiodic processes (phonation onset), irregular vocal fold vibration (paralysis of the recurrent nerve), particular vibration modes (anterior-posterior modes), and running speech. PMID- 11021510 TI - Male reproductive health: cause for concern? AB - A substantial body of evidence has accumulated in recent years that human semen quality may be deteriorating. This has been associated with evidence of other changes in male reproductive health, including increases in congenital malformations and testicular cancer in humans, and similar problems in wildlife. Unfortunately, the evidence remains inconclusive. It has been suggested that these changes may be due to environmental xeno-oestrogens acting during development. Although there is now a large quantity of data indicating that this is a plausible hypothesis, evidence of causality, rather than association, remains to be provided. The potential importance of these changes for human health is considerable and urgent research is required to clarify the situation. PMID- 11021511 TI - Test systems to identify reproductive toxicants. AB - Experience with drugs and other xenobiotics indicates that both animal testing and epidemiological studies are necessary to provide adequate data for an estimation of risks that might be associated with exposure to a chemical substance. In this review, the pros and cons of test systems for reproductive toxicity are discussed. Usually, several studies are performed to cover the different phases of the reproductive cycle. In the preclinical development of drugs, the three so-called 'segment testing protocols' have been used for several decades now. More recently, new testing concepts have been accepted internationally which include more flexibility in implementation. Several examples of compounds with the potential for reproductive toxicity are presented in more detail in a discussion of some pitfalls of the tests for fertility (phthalates and fluoroquinolones), teratogenicity (acyclovir and protease inhibitors) and postnatal developmental toxicity (fluoroquinolones). In addition, important aspects of kinetics and metabolism as a prerequisite for a rational interpretation of results from toxicological studies are briefly discussed. In vitro assays are useful for supplementing the routinely used in vivo approaches or for studying an expected or defined effect, but they are not suitable for revealing an unknown effect of a chemical on the complex reproductive process. PMID- 11021512 TI - In vitro germ cell models for the detection of fertility impairment. AB - Pluripotent embryonic carcinoma cells and pluripotent embryonic stem cells established from undifferentiated cells of an early mouse embryo were investigated for induction of proliferation inhibition, sister chromatid exchanges (SCE) and single-strand breaks by treatment with various germ cell mutagens. The comparison of malignant cells with nonmalignant cells showed an increased sensitivity of nonmalignant cells independent of their state of differentiation. Mitomycin C (MMC) inhibited the proliferation of nonmalignant cells at a concentration of 10(-6) M but did not affect growth of the teratocarcinoma cell line P19. There were no differences between the investigated cell lines at a lower MMC concentration. At the concentration of 10(-6) M MMC the sister chromatid exchanges of P19 were enhanced up to 41 SCE per metaphase. Testing of another germ cell mutagen, ethylnitrosourea (ENU), gave similar results: a decreasing generation time of nonmalignant cell lines after treatment with 1 mM ENU and no effect on the teratocarcinoma cells. This concentration also induced a high number of SCE. Single-strand breaks could be produced by exposure to methanmethylsulphonate (MMS). 56.3% of embryonic stem cell DNA was passing through the filter after MMS treatment. In contrast to the embryonic stem cells, only 35.6% of teratocarcinoma DNA was affected. PMID- 11021513 TI - In vitro tests for essential sperm functions using the phyto-oestrogen genistein as a test substance. AB - Sperm motility, binding of spermatozoa to the zona pellucida and induction of the acrosome reaction are prerequisites for successful oocyte fertilization. Examination of the physiological and nonphysiological effects of particular compounds on sperm functions requires high-quality in vitro test systems. In this short methodological overview, a reliable combined in vitro test system with bovine gametes is described. The purpose of the study was to evaluate whether aliquots of pooled post-thaw spermatozoa are suitable for examination of environmental substances that affect essential sperm functions. The combined test system includes a number of known methods for the assessment of sperm vitality and motion parameters, acrosomal status, inducibility of acrosome reaction and sperm zona pellucida binding. First observations indicate that genistein inhibits the induction of acrosomal exocytosis and binding of spermatozoa to the zona pellucida. Motility parameters and the viability of bovine spermatozoa were not affected by this substance. It is concluded that genistein, a phyto-oestrogen which is abundant in several plants, can be used as a test substance for the evaluation of effects upon essential bovine sperm functions in vitro. PMID- 11021514 TI - Spermatogenesis and mutagenicity of environmental hazards: extrapolation of genetic risk from mouse to man. AB - To perform germ cell mutagenicity studies it is mandatory to know the duration of the different stages of spermatogenesis. The timing of male germ cell development determines the test protocols. Chemical mutagens are characterized by their differential spermatogenic responses, e.g. different chemicals induce mutations in different germ cell stages. Knowledge of the sensitive germ cell stages for a test agent is essential for the evaluation of the genetic hazard, i.e. stem cell effects present permanent genetic hazards and post-stem cell effects present transient hazards. A variety of assays are available to determine germ cell mutagenicity in treated animals or in the progeny of treated animals. Germ cell cytogenetics in differentiating spermatogonia and the dominant lethal assay are used for genetic hazard identification. Their results allow categorization of chemicals as germ cell mutagens (Maximale Arbeitsplatz Konzentration categories for germ cell mutagens). Gene mutations or reciprocal chromosome translocations induced in germ cells are assessed by observation of mutant offspring of treated males. These results are applicable to the quantification of genetic hazards for chemical exposures which cannot be avoided, i.e. for occupational exposures to chemicals such as butadiene. PMID- 11021515 TI - Sertoli cells as a target for reproductive hazards. AB - Male fertility can be impaired by various toxicants. Some of them are known to target mainly Sertoli cells, which play an essential role in spermatogenesis. In this study, the in vitro response of immature rat Sertoli cells to various environmental pollutants, including pesticides, oestrogenic compounds and heavy metals, has been investigated. Mitochondrial dehydrogenase activity has been used to measure Sertoli cell viability, while production of lactate and secretion of inhibin B have been used as general and specific cell markers. Sertoli cell viability was not affected after 24-h exposure to lindane, DDT, ethinyloestradiol or bisphenol A in the concentration range analysed (up to 100, 25 or 50 microM, respectively). In contrast, mercury(II) (EC50 = 31 microM) and cisplatin (15% decrease in viability at 100 microM) induced some cytotoxic effect. With the exception of the pesticide DDT, all chemicals investigated induced a significant dose-dependent increase in lactate production after 24-h exposure to Sertoli cells. Owing to the cytotoxic effect of mercury(II), lactate levels dropped again at concentrations above 20 microM. The pesticide lindane (but not DDT) and both oestrogens significantly increased the production of the Sertoli cell specific hormone inhibin B without affecting cell viability. In contrast, the heavy metals mercury(II) and platinum(II) markedly decreased inhibin B levels. This sharp decrease was already significant at metal concentrations that reduced Sertoli cell viability only moderately (10-15%). In conclusion, the secretion of lactate and inhibin B by immature rat Sertoli cells seems to be a useful and sensitive marker with which to explore potential Sertoli cell toxicants. PMID- 11021516 TI - Evaluation of aneuploidy and DNA damage in human spermatozoa: applications in field studies. AB - With the goal of incorporating measures of sperm nuclear integrity in an epidemiology study, semen samples from young Czech men were analysed for sperm aneuploidy and sperm chromatin structure in addition to routine measures of sperm production and quality. The exposure in question was to high seasonal air pollution containing reactive polyaromatic hydrocarbons potentially capable of affecting spermatogenesis and damaging sperm DNA. The sperm aneuploidy assay uses fluorescence in situ hybridization to label selected sperm chromosomes; as applied in this study, the sex chromosomes (X,Y) and chromosome 8 were targeted. The sperm chromatin structure assay detects sperm nuclei with increased susceptibility to denaturation, a feature that is associated with DNA damage. Logistically, these assays were relatively easy to incorporate into the study design. The aneuploidy assay provided information suggesting that exposure to high levels of air pollution may increase the risk of sperm aneuploidy and that it is important to control for exposure to cigarette smoke and/or alcohol in such studies. The sperm chromatin structure assay provided valuable baseline information about Czech semen donors and data suggestive of an adverse effect of smoking and air pollution on spermatozoa that merits further investigation. PMID- 11021517 TI - Xenobiotic metabolism, genetic polymorphisms and male infertility. AB - Male reproductive function may be impaired by various occupational and environmental chemical agents. The majority of these xenobiotics, however, require metabolic activation in order to exert adverse effects via covalent interactions between intermediate metabolites and cellular macromolecules such as DNA or protein. In addition, metabolization may alter endocrine-disrupting properties of xenobiotics. Thus tissue-specific expression and regulation of multiple xenobiotic-metabolizing enzymes are likely to play an important role in chemically induced disorders of male reproductive organs. Recent studies suggest that genetic polymorphisms underlying inter-individual and inter-ethnic variability of xenobiotic metabolism modulate susceptibility to male reproductive disorders. For cytochrome P450 1A1 (CYP1A1), a key enzyme in extra-hepatic metabolic activation of lipophilic xenobiotics, increased frequencies of two genetically linked polymorphisms have been found among infertile men. PMID- 11021518 TI - Environmental exposure to dioxin: the Seveso experience. AB - The toxicity in humans of 2,3,7,8-tetrachlorodibenzo-p-dioxin, a man-made compound and environmental pollutant, is still debated. The industrial accident at Seveso, Italy, in 1976 exposed a large population of both sexes and of all ages to a massive concentration of 2,3,7,8-tetrachlorodibenzo-p-dioxin. Monitoring of soil and measurement of blood samples allowed classification of the exposed population into three categories: A, B and R (high, medium and low exposure, respectively). This article presents data from longitudinal health monitoring of the population, including liver function, immune function, neurological impairment, dermatological effects, reproductive pathology, and mortality. PMID- 11021519 TI - New methods for detection of potential endocrine disruptors. AB - It has been hypothesized that recent adverse trends in humans are linked to an increased exposure to potential endocrine disrupting agents. These include widely used compounds that mimic the action of sex hormones, including bisphenol A, phthalates and parabens. Since the chemical structure is not sufficient to determine whether a chemical will act as an oestrogen, there is a need for assays that can determine whether a compound interferes with the endocrine systems. The Environmental Protection Agency has recently suggested a testing scheme, composed of an initial screening followed by a more comprehensive investigation of chemicals that are positive in the screening. The screening will use several short-term assays to screen many thousands of compounds for potential endocrine disrupting properties. However, none of these tests determines compound-induced effects on the expression of endogenous genes, which is the cause of the adverse effects. We propose to use a precise quantification of the expression levels of endogenous oestrogen-regulated genes to test whether a chemical has oestrogenic properties, and describe how an endogenous gene expression assay can be established and conducted. Furthermore, different applications of such an assay are discussed: in cell cultures; in experimental animals; or, optimally, directly in blood samples from exposed humans. PMID- 11021520 TI - Receptor-mediated effects of chlorinated hydrocarbons. AB - This short review summarizes some recent findings on arylhydrocarbon (dioxin) receptor expression during early pregnancy in mammals. The arylhydrocarbon receptor is a ligand-activated transcription factor and was originally described as a mediator of the toxic effects of dioxins and other chlorinated hydrocarbons such as polychlorinated biphenyls. Orally administered polychlorinated biphenyls accumulate in uterine secretions, in the pre-implantation blastocyst, in the foetus and in the placenta. Coplanar polychlorinated biphenyls are known ligands of the arylhydrocarbon receptor. Deletion experiments indicate physiological roles of the arylhydrocarbon receptor during development and for the function of various organs. During early pregnancy, the arylhydrocarbon receptor is specifically expressed in pre-implantation embryos, during blastocyst differentiation and implantation, in the endometrial epithelium and in the decidua cells of the placenta. Coplanar polychlorinated biphenyls were found to be embryotoxic in low doses, i.e. in doses found in tissues of not specifically exposed individuals originating from xenobiotic environmental background contamination. In initial experiments, low-dose exposure of rabbit blastocysts to coplanar polychlorinated biphenyls in vitro did not induce transcriptional changes of the so-called arylhydrocarbon receptor gene battery. The embryological and toxicological implications of the findings are discussed. PMID- 11021522 TI - Oestrogen receptor alpha and beta in rat prostate and epididymis. AB - The cellular localization of two oestrogen receptor (ER) subtypes, ER alpha and ER beta, was investigated in neonatal, postnatal, immature and adult male rats to determine whether these receptor subtypes are differentially expressed in prostate and epididymis. A monoclonal antibody against ER alpha and two polyclonal ER beta antibodies were used. Paraffin sections revealed a specific nuclear immunoreaction product in certain cells but not in others. In the epididymis, nuclear ER alpha immunoreactivity (IR) was detected in epithelial cells of efferent ductules and initial segments as well as in connective tissue surrounding the tubules in caput, corpus and cauda. No IR was observed in rete testis. Epithelial cells of the prostate lacked ER alpha IR, but connective tissue cells surrounding prostatic buds in the early neonatal period revealed IR. In prostate, ER beta IR was expressed in epithelial cells of the ventral and dorsolateral lobes, but the IR intensity was higher in the ventral lobe. In neonatal rats, ER beta was expressed in the epididymis but not in the prostate gland. Weak ER beta expression was found in the prostates of 5-day-old rats, and the reaction increased in intensity thereafter. In the epididymis, a similar developmental expression pattern of ER beta was observed. ER beta expression in prostate and epididymis was similar to expression of androgen receptors reported previously for these organs. The results support that both ER alpha and ER beta may be involved in oestrogen modulation of prostate and epididymal functions. PMID- 11021521 TI - Male adolescent exposure to endocrine-disrupting pesticides: vinclozolin exposure in peripubertal rabbits. AB - Adolescence is a time of dramatic neuroendocrine changes that are required for sexual maturation. Hormonal mimicking or inhibiting chemicals can cause significant impairment during this critical period. Vinclozolin (Vin) has been shown to be an anti-androgen affecting male offspring in rats in utero, and its mechanism of action may be mediated by inhibition of androgenic receptor action. The majority of teenagers working on farms are male, and therefore a systemic fungicide, vinclozolin, was selected for study. The rabbit has proved to be an excellent species for modelling reproductive toxicant effects in the male and was selected as the test species. The peripubertal phase for the rabbit was determined to be between the 3rd and 4th months. A 2-month dosing period was therefore initiated at 3 months of age and carried through to the 4th month. Vin was administered by dermal application (100 mg kg(-1) in 100 microl of dimethylsulphoxide) daily. Body weights were determined weekly. The rabbits were then held until fully mature (6 months of age). Semen was collected and evaluated from sexually mature males on a weekly schedule for 5 weeks to maximize sperm output. An automated solid phase extraction procedure for monitoring exposures through isolation and quantification of Vin and its metabolic products was developed. Increased plasma levels of Vin and M2 were found throughout the experimental period. The exposed rabbits had a smaller weight gain during pubertal growth (approaching significance; P=0.059). At maturity, the accessory sex glands of the exposed animals weighed less than those of the controls (P=0.016). Surprisingly, the pooled sperm count of the exposed animals was significantly higher (P=0.017) than that of the unexposed animals. The anti androgenic effects of Vin may have blocked the negative feedback mechanism of testosterone on the hypothalamus or pituitary gland, allowing for an increase in gonadotrophin release, and consequently increasing sperm production at puberty. PMID- 11021523 TI - Effect of two complex platinum salts on human sperm motility and acrosome reaction. AB - Hydrogen hexachloroplatinate, H2PtCl6, has been shown to induce the human sperm acrosome reaction in vitro. However, the molecular mechanism underlying this exocytic process has not been studied. Therefore, two structurally and chemically different platinum (Pt) compounds, the potent sensitizer sodium-hexachloro platinate-(IV), Na2[PtCl6], and the nonimmunogenic tetraamineplatinum-(II) chloride, [Pt(NH3)4]Cl2, were selected for the experiments. Their effects on human sperm function and second messenger pathways were investigated. Washed human spermatozoa were treated with different concentrations of both Pt salts (0.5-1000 microM) during or after capacitation for 3 h at 37 degrees C. In addition, spermatozoa were incubated with Pt salts in calcium-free medium or in the presence of the protein kinase A+C inhibitor H7. Sperm motility was evaluated by computer-assisted sperm analysis; acrosomal loss was detected by triple staining. Compared with the controls (6.6+/-2.4%), the percentages of living acrosome-reacted spermatozoa showed a significant dose-dependent increase (P<0.001) after 3 h of incubation with Na2[PtCl6] (7.9+/-4.2% for 0.5 microM 25.0+/-2.9% for 1 mM) and [Pt(NH3)4]Cl2 (7.9+/-3.9% to 21.0+/-5.8%). Sperm motility was markedly reduced in samples containing the highest concentrations of the Pt salts. The acrosome reaction was also significantly increased when spermatozoa had first been capacitated and then treated with both Pt salts. Calcium-free medium had no effect on the ability of both Pt salts to induce the acrosome reaction. However, incubation of Na2[PtCl6] in the presence of H7 tendentiously decreased the percentage of acrosome-reacted spermatozoa. In conclusion, complex Pt salts such as Na2[PtCl6] or [Pt(NH3)4]Cl2 influence human sperm functions by inducing the acrosome reaction during or after capacitation. This stimulatory effect is independent of calcium and seems to be dependent on protein kinase A or C. PMID- 11021524 TI - Effects of chlorinated hydrocarbons on sperm function in vitro. AB - For the past few years there has been controversial discussion of whether environmental pollutants in general, and chlorinated hydrocarbons in particular, may impair male fertility. Organochlorine compounds, e.g. dichlorodiphenyltrichlorothane (DDT) and metabolites, polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are highly persistent in the environment and there is therefore some concern about human exposure. These chlorinated compounds are universally found in human body fluids. Substantial amounts are detected in human body fluids associated with reproduction such as follicular fluid, seminal fluid and cervical mucus. The available data on male fertility and organochlorines are scarce and controversial. Fertilization rates in in vitro fertilization procedures were found to be decreased in couples in whom the male partner was exposed to pesticides. Data on the effects of PCBs on human sperm motility and acrosome reaction in vitro are controversial. Various PCDD congeners had no effect on human sperm motility in vitro. Effects of chlorinated hydrocarbons on sperm function in vivo seem to be unlikely since the concentrations used in vitro were far higher than those found in fluids of the human reproductive tract. However, negative effects on human spermatozoa in vivo cannot be totally excluded because other organohalogen compounds can be identified in the genital tract and little is known about their synergistic effects. PMID- 11021525 TI - Metal ions and human sperm mannose receptors. AB - Zinc and lead concentrations were measured in seminal plasma from fertile donors, infertile men with varicocoele and men undergoing work-ups for in vitro fertilization. Ejaculated spermatozoa from these subjects were incubated in vitro with various metal ions and/or dibromoethane and dibromochloropropane. Mannose receptor expression was correlated with metal and toxicant levels. Sperm distributions of potassium channels were compared with lead ions and calcium channels with zinc ions. Mannose receptor expression by capacitated spermatozoa increased linearly with seminal plasma zinc levels, and correlated inversely with lead levels. Cobalt had no effect on mannose receptor expression, but nickel had a concentration-dependent biphasic effect. Mannose receptor expression was not affected by dibromoethane and dibromochloropropane if the cholesterol content of the sperm membrane was high, but mannose receptor expression was decreased in low cholesterol spermatozoa by exposures below estimated permissive exposure limits. Potassium channels and lead ions co-localized over the entire head of human spermatozoa, while both calcium channels and zinc ions were confined to the equatorial segment of the head. Mannose receptor expression on the external surface of the human sperm plasma membrane is a biomarker for the effects of transition and heavy metals and organic toxicants on sperm fertility potential. PMID- 11021526 TI - SOCS proteins, regulators of intracellular signaling. PMID- 11021527 TI - Functional flexibility in T cells: independent regulation of CD4+ T cell proliferation and effector function in vivo. AB - Proliferation and differentiation of CD4+ T cells are often correlated, but it is not clear whether they are mechanistically linked. When antigen-specific T cells are present at high frequency in vivo, they all respond to antigenic peptide stimulation by expressing activation markers, but only a subset begins to proliferate. However, noncycling cells may synthesize the effector cytokine IFNgamma even though their cell cycle is blocked in G1. These data show that proliferation and effector function are not rigidly linked in T cells. Instead, CD4+ T cells have the flexibility to engage in or bypass clonal expansion based on the integration of multiple signals, including the frequency of other responding T cells. PMID- 11021528 TI - Soluble CD86 is a costimulatory molecule for human T lymphocytes. AB - CD86 is an important costimulatory molecule for the priming and activation of naive and memory T cells, respectively. Here, we show that soluble CD86 is detected in human serum. Soluble CD86 is produced by resting monocytes and results from an alternatively spliced transcript (CD86deltaTM) characterized by deletion of the transmembrane domain. Recombinant CD86deltaTM binds to CD28 and CTLA-4 and induces the activation of T cells after stimulation with anti-CD3 mAb. CD86deltaTM also induces IFNgamma production by virus-specific CD8+ memory human T cells stimulated with the Flu M1 peptide. The concentrations of soluble CD86 found in human serum are sufficient to induce biological activity. Soluble CD86 molecule, therefore, appears to be a functional costimulatory molecule playing a potentially important role in immune surveillance. PMID- 11021529 TI - The CD28 and CTLA-4 receptors associate with the serine/threonine phosphatase PP2A. AB - CD28 and CTLA-4 are related members of a family of T lymphocyte cell surface receptors that function to regulate T cell activation. We have found that the cytoplasmic domains of both CTLA-4 and CD28 can associate with members of the PP2A family of serine/threonine phosphatases. The association of PP2A with CD28 was negatively regulated by tyrosine phosphorylation of the CD28 cytoplasmic domain. Inhibition of PP2A activity in Jurkat leukemia T cells by treatment with okadaic acid or by expression of a dominant-negative mutant enhanced T cell activation induced by CD28 engagement. Interactions between cell surface receptors such as CTLA-4 and CD28 and serine/threonine phosphatases may represent a novel mechanism for modulating the intracellular signal transduction pathways associated with cell activation. PMID- 11021530 TI - Antigen presentation in extracellular matrix: interactions of T cells with dendritic cells are dynamic, short lived, and sequential. AB - Cognate interactions of naive T cells with antigen-presenting dendritic cells require physical cell-cell contacts leading to signal induction and T cell activation. Using a three-dimensional collagen matrix videomicroscopy model for ovalbumin peptide-specific activation of murine and oxidative mitogenesis of human T cells, we show that T cells maintain vigorous migration upon cognate interactions to DC (dendritic cell), continuously crawl across the DC surface, and rapidly detach (median within 6-12 min). These dynamic and short-lived encounters favor sequential contacts with the same or other DC and trigger calcium influx, upregulation of activation markers, T blast formation, and proliferation. We conclude that a tissue environment supports the accumulation of sequential signals, implicating a numeric or "digital" control mechanism for an ongoing primary immune response. PMID- 11021531 TI - Differences that matter: major cytotoxic T cell-stimulating minor histocompatibility antigens. AB - Despite thousands of genetic polymorphisms among MHC matched mouse strains, a few unknown histocompatibility antigens are targeted by the cytotoxic T cells specific for tissue grafts. We isolated the cDNA of a novel BALB.B antigen gene that defines the polymorphic H28 locus on chromosome 3 and yields the naturally processed ILENFPRL (IFL8) peptide for presentation by Kb MHC to C57BI/6 CTL. The CTL specific for the IFL8/Kb and our previously identified H60/Kb complexes represent a major fraction of the B6 anti-BALB.B immune response. The immunodominance of these antigens can be explained by their differential transcription in the donor versus the host strains and their expression in professional donor antigen-presenting cells. PMID- 11021532 TI - DAP12-deficient mice fail to develop autoimmunity due to impaired antigen priming. AB - DAP12 is an ITAM-bearing membrane adaptor molecule implicated in the activation of NK and myeloid cells. In mice rendered DAP12 deficient by targeted gene disruption, lymphoid and myeloid development was apparently normal, although the activating Ly49 receptors on NK cells were downregulated and nonfunctional. To analyze the consequences of DAP12 deficiency in vivo, we examined the susceptibility of DAP12-/- mice to experimental autoimmune encephalomyelitis (EAE). DAP12-/- mice were resistant to EAE induced by immunization with myelin oligodendrocyte glycoprotein (MOG) peptide. Resistance was associated with a strongly diminished production of IFNgamma by myelin-reactive CD4+ T cells due to inadequate T cell priming in vivo. These data suggest that DAP12 signaling may be required for optimal antigen-presenting cell (APC) function or inflammation. PMID- 11021533 TI - Combined natural killer cell and dendritic cell functional deficiency in KARAP/DAP12 loss-of-function mutant mice. AB - KARAP/DAP12 is a transmembrane polypeptide with an intracytoplasmic immunoreceptor tyrosine-based activation motif (ITAM). KARAP/DAP12 is associated with several activating cell surface receptors in hematopoietic cells. Here, we report that knockin mice bearing a nonfunctional KARAP/DAP12 ITAM present altered innate immune responses. Although in these mice NK cells are present and their repertoire of inhibitory MHC class I receptors is intact, the NK cell spectrum of natural cytotoxicity toward tumor cell targets is restricted. KARAP/DAP12 loss-of function mutant mice also exhibit a dramatic accumulation of dendritic cells in muco-cutaneous epithelia, associated with an impaired hapten-specific contact sensitivity. Thus, despite its homology with CD3zeta and FcRgamma, KARAP/DAP12 plays a specific role in innate immunity, emphasizing the nonredundancy of these ITAM-bearing polypeptides in hematopoietic cells. PMID- 11021534 TI - Inhibition of natural killer cell-mediated cytotoxicity by Kaposi's sarcoma associated herpesvirus K5 protein. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) K3 and K5 proteins dramatically downregulate MHC class I molecules. However, although MHC class I downregulation may protect KSHV-infected cells from cytotoxic T lymphocyte recognition, these cells become potential targets for natural killer (NK) cell-mediated lysis. We now show that K5 also downregulates ICAM-1 and B7-2, which are ligands for NK cell-mediated cytotoxicity receptors. As a consequence, K5 expression drastically inhibits NK cell-mediated cytotoxicity. Conversely, de novo expression of B7-2 and ICAM-1 resensitizes the K5-expressing cells to NK cell-mediated cytotoxicity. This is a novel viral immune evasion strategy where KSHV K5 achieves immune avoidance by downregulation of cellular ligands for NK cell-mediated cytotoxicity receptors. PMID- 11021535 TI - Structure of the human IgE-Fc C epsilon 3-C epsilon 4 reveals conformational flexibility in the antibody effector domains. AB - IgE antibodies mediate antiparasitic immune responses and the inflammatory reactions of allergy and asthma. We have solved the crystal structure of the human IgE-Fc Cepsilon3-Cepsilon4 domains to 2.3 A resolution. The structure reveals a large rearrangement of the N-terminal Cepsilon3 domains when compared to related IgG-Fc structures and to the IgE-Fc bound to its high-affinity receptor, FcepsilonRI. The IgE-Fc adopts a more compact, closed configuration that places the two Cepsilon3 domains in close proximity, decreases the size of the interdomain cavity, and obscures part of the FcepsilonRI binding site. IgE-Fc conformational flexibility may be required for interactions with two distinct IgE receptors, and the structure suggests strategies for the design of therapeutic compounds for the treatment of IgE-mediated diseases. PMID- 11021536 TI - Crystal structure of the extracellular domain of a human Fc gamma RIII. AB - Fc receptors play a major role in immune defenses against pathogens and in inflammatory processes. The crystal structure of a human immunoglobulin receptor, FcgammaRIIIb, has been determined to 1.8 A resolution. The overall fold consists of two immunoglobulin-like domains with an acute interdomain hinge angle of approximately 50 degrees. Trp-113, wedged between the N-terminal D1 and the C terminal D2 domains, appears to further restrict the hinge angle. The putative Fc binding region of the receptor carries a net positive charge complementary to the negative-charged receptor binding regions on Fc. A 1:1 binding stoichiometry between the receptor and Fc was measured by both the equilibrium and nonequilibrium size-exclusion chromatography. Two separate parallel dimers are observed in the crystal lattice, offering intriguing models for receptor aggregation. PMID- 11021537 TI - Abnormal Stat activation, hematopoietic homeostasis, and innate immunity in c-fes /- mice. AB - The c-fes protooncogene encodes a nonreceptor tyrosine kinase (Fes) implicated in cytokine receptor signal transduction, neutrophil survival, and myeloid differentiation. To determine the role of Fes in embryonic development and hematopoiesis, we engineered a null mutation of the murine c-fes locus. c-fes-/- mice are viable but not born in the expected Mendelian ratios. Live born c-fes-/- mice exhibit lymphoid/myeloid homeostasis defects, compromised innate immunity, and increased Stat activation in response to GM-CSF and IL-6 signaling. Therefore, increased cytokine responsiveness in the absence of Fes leads to abnormal myeloid proliferation and functional defects in the macrophage lineage. PMID- 11021538 TI - The contribution of somatic hypermutation to the diversity of serum immunoglobulin: dramatic increase with age. AB - Although somatic mutation contributes to the diversity of only a minor fraction of B cells in mouse spleen or blood, its contribution to the diversity of serum immunoglobulin is unknown. We have devised an immunoassay to monitor mutated antibodies in serum using a monoclonal antibody that recognizes a VK only when mutated at its major intrinsic hot spot. Mutation makes essentially no contribution to the diversity of endogenous serum IgM, IgG, or IgA in young mice. However, in response to environmental antigens, the titer of mutated immunoglobulin in T cell-proficient mice rises strikingly with age, such that the major proportion of serum immunoglobulin in adults is somatically mutated, with the mutation load in IgG being some 10-fold greater than in IgM. PMID- 11021539 TI - Development of computational and graphical tools for analysis of movement and flexibility in large molecules. AB - We developed a computer program for the calculation and display of the difference distance matrices (DDMs) of macromolecules that has the ability to compare multiple structures simultaneously. To demonstrate its use, a data set of atoms for superimposition of the HIV-1 reverse transcriptase enzyme was defined using the coordinates for the 21 available crystal structures of this enzyme and its complexes. The DDM technique for superimposition data set generation allows selection of atoms that are invariant in all structures, is free from user bias, and represents the most accurate and precise method of producing such subsets. Comparison of this technique was made against other published methods of generating superimposition data sets, and it was found that significant differences in magnitude and trends of atom movements are observed depending on which data set was used. PMID- 11021540 TI - An algorithm for predicting the NMR shielding of protons over substituted benzene rings. AB - In a strong magnetic field, hydrogen nuclei located over an aromatic ring experience a reduced magnetic field as a result of the induced magnetic field associated with circulating pi electrons. We used GIAO-SCF, an ab initio subroutine in Gaussian 94 to calculate isotropic shielding values and to determine the proton nuclear magnetic resonance (NMR) shielding increment for a simple model system: methane held at various positions over a substituted benzene ring. The NMR shielding increments experienced by the proximal protons of methane have been mapped as a function of their position X, Y, and Z relative to the center of aniline and, separately, nitrobenzene. A mathematical function of the same form has been fit to the three-dimensional shielding increment surface at each of five distances from the face of each aromatic ring. In addition, a single mathematical equation has been developed for predicting the shielding caused by either substituted aromatic ring. The chemical shifts predicted by using the results of this equation in conjunction with additive substituent increments are compared to observed values. PMID- 11021541 TI - A method for including protein flexibility in protein-ligand docking: improving tools for database mining and virtual screening. AB - Second-generation methods for docking ligands into their biological receptors, such as FLOG, provide for flexibility of the ligand but not of the receptor. Molecular dynamics based methods, such as free energy perturbation, account for flexibility, solvent effects, etc., but are very time consuming. We combined the use of statistical analysis of conformational samples from short-run protein molecular dynamics with grid-based docking protocols and demonstrated improved performance in two test cases. Our statistical analysis explores the importance of the average strength of a potential interaction with the biological target and optionally applies a weighting depending on the variability in the strength of the interaction seen during dynamics simulation. Using these methods, we improved the num-top-ranked 10% of a database of drug-like molecules, in searches based on the three-dimensional structure of the protein. These methods are able to match the ability of manual docking to assess likely inactivity on steric grounds and indeed to rank order ligands from a homologous series of cyclooxygenase-2 inhibitors with good correlation to their true activity. Furthermore, these methods reduce the need for human intervention in setting up molecular docking experiments. PMID- 11021542 TI - An automatic homology modeling method consisting of database searches and simulated annealing. AB - We introduce a method of homology modeling consisting of database searches and simulated annealing. All processes involving searches for homologous proteins, alignment, the construction of Calpha atoms, construction of main-chain atoms, and the construction of side-chain atoms are performed automatically. In this method, main-chain conformations are generated from the weighted average of mainchain coordinates in reference proteins. The weight is defined by the local space homology representing the similarity of environmental residues at topologically equivalent positions in reference proteins. Side-chain conformations are generated for constructed main-chain atoms by database searches, and main-chain atoms are optimized for the fixed side-chain conformations. These two processes, i.e., the side-chain generation and main chain optimization, are repeated several times. This type of construction provides a structure similar to the x-ray structure, in particular, for main chain and side-chain atoms in the residues belonging to structurally conserved regions (SCRs). The accuracy of our method was evaluated for 14 proteins whose structures are known. The average root mean square deviation between models and x ray structures was 2.29 A for all atoms, and the percentage of chi1 angles within 30 degrees was 72.6% for SCRs residues. Some models were in good agreement with their respective x-ray structures. Our method, which has the advantage of being automated, gives results similar to, or better than, published results for three widely used test proteins. Our software, FAMS, is available on the World Wide Web. PMID- 11021543 TI - GB/SA water model for the Merck molecular force field (MMFF). AB - A revised generalized Born/surface area (GB/SA) continuum solvation model has been developed for water that is compatible with the Merck molecular force field (MMFF). This model gives free energies of aqueous solvation that are comparable in accuracy to the original water model when the OPLS* force field is employed. The average unsigned error in aqueous deltaGsol using the new water model and MMFF is 0.62 kcal/mol for a training set of 82 solutes compared to 1.24 kcal/mol for the original GB/SA water model and MMFF. The average unsigned errors for 47 neutral solutes outside the training set and 10 ions are 0.96 and 2.32 kcal/mol, respectively. By comparison, the average errors for the test set and ions using the original GB/SA water model are 1.76 and 5.32 kcal/mol. This revised parameter set provides a more accurate representation of aqueous solvation for use with MMFF. PMID- 11021544 TI - Docking of sulfonamides to carbonic anhydrase II and IV. AB - Starting with a known active site of a protein and a database of compounds, one would like to quickly identify a few compounds that "dock" into the active site and obtain "good" binding free energies. The main goal of current automated docking procedures is to predict the "best" substrate-enzyme complex while other programs such as UHBD and DelPhi can be used to compute binding free energies. In this paper, we will focus on the application of docking methods and parameters to study substrate-enzyme interactions of a metalloenzyme system. Specifically, we report on the docking of sulfonamides to carbonic anhydrase II and IV, which are of interest due to their application in glaucoma therapy. Using a standard docking protocol, it is possible to correctly predict not only the orientation of inhibitors to a specific isozyme, but also determine the qualitative affinity for a group of inhibitor for an isozyme. PMID- 11021545 TI - SymROP: ROP protein with identical helices redesigned by all-atom contact analysis and molecular dynamics. AB - Experience has shown that protein redesigns (using the backbone from a known protein structure) are far more likely to produce well-ordered, native-like structures than are true de novo designs. Therefore, to design a four-helix bundle made of identical short helices, we here proceed by an extensive redesign of the ROP protein. A fully symmetrical SymROP sequence derived from ROP was chosen by modeling ideal-geometry side chains, including hydrogens, while maintaining the "goodness-of-fit" of side-chain packing by calculating all-atom contact surfaces with the Reduce and Probe programs. To estimate the probable extent of backbone movement and side-chain mobility, restrained molecular dynamics simulations were compared for candidate sequences and controls, including substitution of Abu for all or half the core Ala residues. The resulting 17-residue designed sequence is 41% identical to the relevant regions in ROP. SymROP is intended for construction by the Template Assembled Synthetic Proteins approach, to control the bundle topology, to use short helices, and to allow blocked termini and unnatural amino acids. ROP protein has been a valuable system for studying helical protein structure because of its simplicity and regularity within a structure large enough to have a real hydrophobic core. The SymROP design carries that simplicity and regularity even further. PMID- 11021546 TI - Part VI. Toxic inhalants. PMID- 11021547 TI - Part VII. Air pollution: overview of sources and health effects. AB - In this era of increasing interest about environmental contributors to illness, primary care providers may be asked by patients for guidance about air pollution. Some components of outdoor air pollution have improved in the past 30 years, but locally hazardous conditions continue to occur, especially for vulnerable subpopulations. Such groups include people with chronic cardiac and respiratory conditions, the elderly, pregnant women, and newborns. This overview discusses the sources and health effects of the primary pollutant groups--ozone, sulfur dioxide (SO2), nitrogen oxides, carbon monoxide (CO), particulates, and other air toxics. A number of Web links are provided to help clinicians and patients get up to-date information about pollution in the areas where they live. This section concludes with a discussion of treatment and of behavioral and societal approaches to air pollution. PMID- 11021548 TI - Part VIII. Identification and control of selected residential hazards. PMID- 11021549 TI - Part IX. Laboratory evaluation of low-level chemical or toxic exposures. PMID- 11021550 TI - Perspectives on the use of frequency doubling and short wavelength perimetry for the diagnosis of glaucoma. PMID- 11021551 TI - A comparison of short wavelength automated perimetry with frequency doubling perimetry for the early detection of visual field loss in ocular hypertension. AB - BACKGROUND: Achromatic automated perimetry (AAP) is limited in its ability to detect very early visual field loss in ocular hypertensive patients. Tests targeting axons that are selectively damaged, or have low redundancy, may detect visual field losses before they are seen on AAP. It has been claimed that short wavelength automated perimetry (SWAP) and frequency doubling perimetry (FDP) are two tests that provide early detection. METHODS: Patients (n = 62) were selected on the basis that they had raised intraocular pressure but normal visual fields detected by AAP. A SWAP and an FDP was performed on each of the patients and the results compared. Fields were scored as either normal or abnormal based on criteria used in previous studies. RESULTS: On comparing FDP with SWAP as the 'gold standard', a sensitivity of 88.9% and a specificity of 96.2% was found, showing a high concordance between the two tests. CONCLUSION: These results suggest that as SWAP may be predictive of AAP visual field loss, FDP may be similarly predictive. PMID- 11021552 TI - Ocular parameters in the subgroups of angle closure glaucoma. AB - PURPOSE: This study was conducted to compare anatomical parameters, thought to be responsible for causing angle closure glaucoma (ACG), among eyes having acute, subacute or chronic ACG. METHODS: Ninety consecutive patients diagnosed with a subgroup of ACG, and 30 age, sex and refraction matched controls, provided a total of 240 eyes for a prospective study. The refractive error, corneal diameter, keratometry, pachymetry, lens thickness and axial length were measured and the relative lens position was calculated. The data were analysed by paired t test, ANOVA, signed rank test and multivariate analysis. RESULTS: Acute ACG eyes were mildly hyperopic. All the ACG subgroups had similarly short eyeballs and a steeper corneal curvature compared to control eyes. Acute ACG lenses were thicker than all the other groups (P < 0.001), but all ACG eyes had thicker lenses than the controls. Corneal diameters and anterior chamber depths were decreased in acute and chronic ACG eyes compared with subacute ACG and controls (P < 0.001). The uninvolved fellow eyes in each subgroup differed from affected eyes only in having more posteriorly positioned lenses. CONCLUSIONS: There was a spectrum of anatomical variations seen in the subgroups of ACG. Acute ACG eyes expressed an extreme shift of anatomical features away from normal, especially, smaller corneal diameters, leading to a large mobile lens in an already crowded anterior segment. This predisposed them to a severe relative pupillary block, and to a form of ciliary block glaucoma. Chronic ACG eyes were less divergent from normal and therefore could have suffered a milder form of the same kind of angle closure, but over a more prolonged period. Subacute ACG eyes deviated least from controls, and therefore exhibited mild signs and spontaneous resolution. Further work is required to elucidate completely the pathophysiology that leads to ACG. PMID- 11021553 TI - Beta radiation as an adjunct to low-risk trabeculectomy. AB - PURPOSE: To assess a single dose of intraoperative beta radiation used to enhance the success rate of trabeculectomy in a population of low-risk glaucoma patients in whom antimetabolites might not be indicated. METHODS: A prospective randomized trial of 65 eyes was designed, with 31 eyes receiving 750 rads of intraoperative beta radiation (group 1), and 34 eyes receiving no supplementation (group 2). RESULTS: Mean follow-up time was 24 months. Mean postoperative intraocular pressure was 12.2 mmHg in group 1, and 13.7 mmHg in group 2 (P = 0.16). Mean decrease in intraocular pressure was 10.3 mmHg in group 1, and 9.3 mmHg in group 2 (P = 0.49). The two groups were not significantly different in terms of surgical complications. CONCLUSION: For this population of low-risk patients, there was no significant difference in outcome after applications of a single intraoperative dose of beta radiation. PMID- 11021554 TI - Diode laser cyclophotocoagulation: longer term follow up of a standardized treatment protocol. AB - BACKGROUND: Diode laser cyclophotocoagulation (cyclodiode) is gaining acceptance as a therapy for refractory or end-stage glaucoma. The aim of the study was to establish whether a reproducible dose-effect relationship exists following a single standardized cyclodiode treatment to lower intraocular pressure (IOP). METHODS: This paper reports results from a longer-term follow up of a trial involving the study of 30 patients with uncontrolled IOP and advanced glaucoma who were treated with a single episode of either a half or a full standardized dose of laser depending on clinical risk of phthisis and monitored for IOP control, visual acuity, postoperative inflammation and phthisis. RESULTS: A lowering of IOP at the final follow up was achieved in 83% of patients, with a mean follow-up time of 21 months. For patients receiving a full standardized laser dose, preoperative IOP was 49.4 +/- 11.2 mmHg (mean +/- SD) and postoperative IOP was 28.5 +/- 20.0 mmHg (42% reduction); 45% of patients achieved IOP < 22 mmHg and 68% gained an IOP reduction > 30%. For the half treatment cases, the preoperative IOP was 29.4 +/- 4.3 mmHg (mean +/- SD) and the postoperative IOP was 18.3 +/- 10.0 mmHg (38% reduction); 63% of patients achieved IOP < 22 mmHg and 50% gained an IOP reduction of > 30%. Of 22 sighted eyes, nine (41%) recorded no change in vision; and nine (41%) lost and four (18%) gained vision. Hypotony was seen in 4/22 (18%) of full standardized laser dose cases. CONCLUSION: Longer follow-up times confirm that diode laser cyclophotocoagulation is a convenient and useful therapy in the control of IOP in end-stage glaucoma. Response of IOP to the laser therapy is highly variable, particularly in the neovascular glaucoma group, and it does not appear to be possible to predict an IOP outcome for an individual eye. Circumferential treatments in neovascular eyes should be avoided. Prospects for long-term vision retention in end-stage eyes are poor, perhaps due to progression of the underlying disease. PMID- 11021555 TI - Age-specific prevalence and causes of bilateral and unilateral visual impairment in older Australians: the Blue Mountains Eye Study. AB - PURPOSE: To report age-specific prevalence and causes of bilateral and unilateral visual impairment in a representative older Australian population. METHODS: Participants in the Blue Mountains Eye Study (n = 3654) underwent a detailed eye examination. Any visual impairment was defined as best-corrected visual acuity (VA) of 6/12 or worse, moderate impairment as VA 6/24-6/60 and severe impairment as VA worse than 6/60 (the Australian definition for legal blindness). Bilateral visual impairment was defined from the better eye and unilateral impairment from the worse eye. Proportional causes for visual loss were determined by the examining ophthalmologist. RESULTS: Prevalence of bilateral and unilateral visual impairment was strongly age related. Corresponding bilateral and unilateral visual impairment prevalence rates were, respectively, 0.6% and 3.6% for persons aged 49-59 years, 1.1% and 8.2% for ages 60-69, 5.4% and 20.1% for ages 70-79, and 26.3% and 52.2% for persons aged 80+ years. Overwhelmingly, age-related maculopathy (ARM) was the predominant cause of bilateral blindness (13/17) and of moderate to severe bilateral visual impairment in persons aged 70+ years. However, cataract was the most frequent cause of mild bilateral visual impairment among persons aged 60+ years. Amblyopia was the most frequent cause of mild or worse unilateral visual impairment in persons aged 49-59 years. Cataract was the most common cause of mild unilateral visual impairment in people aged 60+ years, while ARM and cataract were jointly the most frequent causes of moderate to severe unilateral visual impairment in people aged 70+ years. CONCLUSIONS: These findings indicate that around half of visually impaired persons aged 60 years or over had cataract, a cause amenable to treatment. PMID- 11021556 TI - An innovation in developing world cataract surgery: sutureless extracapsular cataract extraction with intraocular lens implantation. AB - PURPOSE: The benefits of sutureless cataract surgery have not been available to the majority of developing world patients. This report describes a surgical technique developed to gain the benefits of self-sealing sutureless techniques, without the need for costly, complex instrumentation, expensive consumables, or difficult nucleus fragmentation manoeuvres in the anterior chamber. METHOD: A detailed description of a sutureless technique that uses only standard manual extracapsular cataract extraction instrumentation is presented. The clinical records of 362 consecutive completed sutureless surgeries were reviewed, and preliminary indicative data of visual outcome for those having follow up between postoperative weeks 3 and 8 are also presented. RESULTS: Hospital and eye camp experience confirms this operation offers speed, routinely 5 min or less with mature cataracts, reduced cost because no suture, viscoelastic or anterior chamber maintainer is needed, wound security and stability, and rapid effective visual rehabilitation. Uncorrected distance visual acuity during the third to eighth postoperative week was available for 266 of the 362 patients (73%): 10, 145, 101, and 10 had acuities of better than 6/9, 6/9-6/18, 6/24-6/60, and worse than 6/60, respectively. Ninety-seven, 165, 12, and two of the 276 (76%) patients with recorded corrected distance acuity were in the same groups. Comparing postoperative with preoperative corneal astigmatism for each patient with postoperative information available (208), without regard to axis, the amplitude of astigmatism worsened by a mean of 0.94 dioptres (+/- 0.17, 95% confidence; +/- 1.23 standard deviation; 7.5D maximum worsening; 2.5D maximum improvement). No attempt has been made to analyse complication type or frequency in this preliminary report. CONCLUSION: The authors believe this sutureless cataract extraction technique to be an innovation with widespread application in the developing world. Further and more exhaustive critical appraisal is warranted. PMID- 11021557 TI - Visually significant calcification of hydrogel intraocular lenses necessitating explantation. AB - PURPOSE: To report two cases of HydroviewR (Storz H60M) hydrogel intraocular lens (IOL) opacification necessitating explantation. To analyse the explanted lenses and compare them with a new lens in order to determine the cause of opacification. METHODS: Two cases are presented. Both patients complained of subjective decreases in vision approximately 1 year following uneventful cataract extraction and implantation of hydrogel lenses by the same experienced surgeon in the same institution. The reduced contrast sensitivity was thought to be due to failure of the hydrogel lenses which appeared opacified at the slit-lamp. They were then exchanged for acrylic lenses. Pre- and postoperative contrast sensitivity testing was performed using the VectorvisionR CSV-1000E instrument. A detailed comparative analysis of the opacified explanted lenses and a new lens was undertaken. This included surface microscopy, infrared spectral analysis, X ray elemental analysis, scanning electron microscopy, and electron microscopy of lens slices. RESULTS: Lens exchange resulted in an improvement in subjective visual acuity (VA) and in objective contrast sensitivity testing. Analysis revealed deposits of calcium phosphate (hydroxyapatite) on the explanted lens' surfaces. CONCLUSIONS: The surface of the HydroviewR H60M intraocular lens is subject to opacification as a result of calcium phosphate (hydroxyapatite) deposition, thereby limiting the patient's visual outcome, and in some cases necessitating explantation. The reason for surface calcification requires further study. PMID- 11021558 TI - Topical anaesthesia for phacoemulsification surgery. AB - AIM: To assess the discomfort experienced by patients undergoing phacoemulsification under topical anaesthesia. METHODS: Thirty patients had a collagen contact lens, soaked in 4% lignocaine, placed on the cornea 23-89 min preoperatively. Immediately preoperatively, additional ophthetic drops were inserted. The patients underwent phacoemulsification surgery with the insertion of a foldable intraocular lens. This group was compared with 30 age- and sex matched controls who underwent phacoemulsification and the insertion of a non foldable lens under peribulbar anaesthesia. (8 mL of an 80:20 mix of 0.5% bupivacaine, 4% lignocaine and hyaluronidase). The pain perceived by patients was assessed on a verbal analogue scale (0-10) at administration of anaesthesia, perioperatively and 3-6 h postoperatively. RESULTS: Topical anaesthesia was less painful than peribulbar anaesthetic at administration (difference in mean pain score 1.2, P < 0.05, Wilcoxon paired signed rank test). Patients experienced more pain during the operation under topical anaesthesia (difference in means 0.63, P < 0.05). There was no difference between the groups postoperatively. CONCLUSION: Topical anaesthesia does not carry the risk of injection into, or injury of, the many delicate structures of the orbit. Because it provides acceptable levels of anaesthesia for phacoemulsification, it could be used more frequently. PMID- 11021559 TI - Effect of lens style on postoperative refractive astigmatism after small incision cataract surgery. AB - PURPOSE: To evaluate the astigmatic results of two popular foldable intraocular lenses when standard surgical approaches are used in rural private practice in Australia. METHODS: Four hundred consecutive cataract operations were followed in a prospective study. The refractive results of patients receiving the Alcon Acrysof MA30 lens were compared to those receiving the Allergan silicone S140 lens. All patients were over 60 years of age and had no intercurrent ocular disease. Refractive and keratometric astigmatism was analysed both by vector analysis and as an absolute measure. RESULTS AND CONCLUSION: The Acrysof lens is associated with less postoperative astigmatism (P = 0.01), than the Silicone S140 lenses. PMID- 11021560 TI - Eyelid retractor surgery as an adjunct to cicatricial ectropion repair. AB - PURPOSE: To evaluate the effectiveness of eyelid retractor repair in cicatricial ectropion of the lower eyelid. METHODS: The study design was a prospective case series. One hundred and twenty eight eyelids were operated on in 100 consecutive patients with cicatricial ectropion. All patients underwent lower eyelid retractor repair via a conjunctival approach combined with skin replacement to the anterior lamella with or without a horizontal lid tightening procedure. When only medial ectropion was present, a medial-based transpositional skin flap was used to repair the anterior lamella (26 eyelids). The remaining eyelids with ectropion involving all or most of the eyelid underwent upper-to-lower eyelid lateral-based transpositional skin flap repair (92 eyelids), or full thickness free skin grafting (10 eyelids). Horizontal lid tightening was performed by lateral canthoplasty in 123 eyelids. RESULTS: Relief of cicatricial ectropion symptoms was reported in 90% of patients overall. A normal punctum position was achieved in 70% of eyelids, overall, and was highest (88%) with a medial-based transpositional skin flap. CONCLUSIONS: Eyelid retractor repair combined with skin replacement and horizontal lid shortening is an effective procedure for cicatricial ectropion. PMID- 11021561 TI - The macular hole: report of an Australian surgical series and meta-analysis of the literature. AB - PURPOSE: To report an Australian series of macular hole surgery by pars plana vitrectomy and fluid-gas exchange, and to identify factors influencing the outcome of the surgery through meta-analysis of the literature. METHODS: Fifty six consecutive cases of macular hole were treated by pars plana vitrectomy, fluid-gas exchange and face-down positioning for at least 7 days, and prospectively followed for 3-12 months. Thirty-six reports of macular hole surgery were reviewed. A meta-analysis on the pre- and postoperative parameters was performed on 389 cases, in which case-specific data-points were available. RESULTS: In the current series, anatomical closure was achieved in all (100%) of 16 stage 2, and in 35 (87.5%) of 40 stage 3 or 4 macular holes. At least 2 logMAR lines of improvement in visual acuity were seen in 10 (62.5%) stage 2 and 20 (50.0%) stage 3 or 4 holes. Postoperative visual acuity was 6/12 or better in 10 (62.5%) stage 2 and 17 (42.5%) stage 3 or 4 holes. In both the current series and the meta-analysis, favourable surgical outcomes were associated with stage 2 macular holes, better preoperative visual acuity, and shorter preoperative duration. Adjuvant use was associated with a higher rate of anatomical closure but there was no clear benefit in terms of postoperative visual acuity. CONCLUSION: Our experience and the results of the meta-analysis suggest that macular hole surgery should be offered as early as possible once full-thickness neuroretinal defect occurs. PMID- 11021562 TI - Central serous retinopathy complicating systemic lupus erythematosus: a case series. AB - Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder with widespread manifestations including the eye. Central serous retinopathy (CSR) has been associated as a complicating event in SLE, although it is uncommon. We present a case series of four female Chinese SLE patients who developed CSR during the course of their systemic disease. All four presented clinically with typical CSR. Angiographic findings did not show evidence of choroidal ischaemia or delayed choroidal filling. Resolution of the serous retinal detachment occurred in all four patients. Recovery of vision was seen in three patients. The clinical outcome was similar to that occurring in the usual male population. Central serous retinopathy as a manifestation of SLE may be caused by various factors. These include SLE-associated choroidopathy, systemic hypertension, renal disease, retinal pigment epithelial dysfunction and glucocorticoid therapy. PMID- 11021563 TI - Congenital porphyria with necrotizing scleritis in a 9-year-old child. AB - Porphyria is a rare metabolic disorder that is characterized by the accumulation of photosensitive, toxic intermediates of the heme metabolic pathway in various organs of the body including the skin, eye and neural tissue. Porphyria as a potential cause for bilateral necrotizing scleritis in children is very infrequently emphasized in literature, probably due to the relatively rare occurrence and lack of well-documented cases. A case of a 9-year-old child with congenital porphyria who had developed necrotizing scleral ulceration in both eyes in addition to severe cutaneous hypersensitivity and facial disfigurement is herein presented. PMID- 11021564 TI - Ocular decompression retinopathy after resolution of acute primary angle closure glaucoma. AB - A patient presented with acute primary angle closure glaucoma with markedly elevated intraocular pressure. Two weeks after laser peripheral iridotomy and resolution of the acute attack, the patient was noted to have developed scattered retinal haemorrhages. The haemorrhages resolved over time with no visual sequelae. This is the first reported case of ocular decompression retinopathy after resolution of acute primary angle closure glaucoma. PMID- 11021565 TI - Indocyanine green angiographic features of varix of the vortex vein ampulla. AB - Varix of the vortex vein ampulla is a rare, benign, asymptomatic condition, which may be confused with a choroidal naevus or melanoma. A 28-year-old man was referred to a tertiary retinal practice with a diagnosis of choroidal naevus. The lesion was an elevated choroidal mass in the superonasal peripheral retina measuring 2 by 1 disc diameters. It was dark red to burgundy in colour and disappeared under digital pressure applied to the globe. The methods used in diagnosis were colour fundus photography, fluorescein angiography and indocyanine green angiography using a scanning laser ophthalmoscope. On fluorescein angiography the lesion was initially hypofluorescent, becoming isofluorescent 25 s after dye injection. Indocyanine green angiography demonstrated the lesion to be two separate dilatations of the vortex vein ampullae. The dilatations collapsed when pressure was applied to the globe. A choroidal mass that collapses under pressure applied to the globe should suggest a varix of the vortex vein ampulla. Indocyanine green angiography is useful in demonstrating the outline of the varix of the vortex vein ampulla. PMID- 11021566 TI - Sino-orbital fistula: two case reports. AB - A fistula between the paranasal sinuses and the orbit as a late complication of orbital fractures is rare and may present with intermittent symptoms due to air passing into the orbit. A case note review of two patients with sino-orbital fistula is presented. Two patients, 23- and 30-year-old males, presented with intermittent symptoms of globe displacement, diplopia or discomfort months after repair of an orbital floor fracture with a synthetic orbital floor implant. The symptoms occurred after nose blowing. They were both cured by removal of the implant and partial removal of the tissue surrounding the implant. A sino-orbital fistula may complicate the otherwise routine repair of an orbital floor fracture, but may be cured by removal of the implant and part of the surrounding pseudocapsule. PMID- 11021567 TI - Bilateral choroidal infarction in a patient with antiphospholipid syndrome: a case report. AB - PURPOSE: To report a case of bilateral choroidal infarction occurring as a complication of primary antiphospholipid syndrome. METHODS: The case notes of the patient were reviewed. RESULTS: A 25-year-old man with primary antiphospholipid syndrome and a previous history of deep vein thrombosis and pulmonary embolism, developed episcleritis and bilateral choroidal infarction, with deterioration of vision to 6/12 OD and counting fingers at 5 feet OS. Anticoagulation therapy with warfarin was administered and the patient's vision gradually improved to 6/6 OD and 6/9 OS. CONCLUSION: Primary antiphospholipid syndrome is associated with thrombotic phenomena, which may affect the ocular arterial and venous vasculature. Therefore, a detailed ophthalmological examination is warranted in those patients who present with ocular symptoms and deteriorating vision. Retinal fluorescein angiography is valuable in confirming the diagnosis when in doubt, and in determining the extent of vessel occlusion. PMID- 11021568 TI - Restoration of ocular circulation and some visual function following external carotid endarterectomy. PMID- 11021569 TI - The incidence of killer activity of non-Saccharomyces yeasts towards indigenous yeast species of grape must: potential application in wine fermentation. AB - Fourteen killer yeasts were assayed for their ability to kill species of yeast that are commonly associated with fermenting grape must and wine. A total of 147 of a possible 364 killer-sensitive interactions were observed at pH 4.5. Of the killer yeasts studied, Pichia anomala NCYC 434 displayed the broadest killing range. At a pH value comparable with those of wine ferments, pH 3.5, the incidence of killer-sensitive interactions was reduced by 700% across all the yeasts. Williopsis saturnus var. mrakii CBS 1707 exhibited the broadest killing range at the lower pH, killing more than half of the tester strains. Intraspecific variation in sensitivity to killer yeasts was observed in all species where more than one strain was tested. Also, in strains of Pichia anomala, Kluyveromyces lactis and Pichia membranifaciens, the three species in which more than one killer yeast was analysed, intraspecific variation in killer activity was observed. PMID- 11021570 TI - A 210-min solid phase cytometry test for the enumeration of Escherichia coli in drinking water. AB - A 210-min-test for the enumeration of Escherichia coli in drinking water is described, based on solid phase cytometry (SPC) and a two-step enzymatic procedure for fluorescence labelling of single cells and small microcolonies. The test involves membrane filtration through a 25-mm black polyester filter, induction of beta-glucuronidase in the retained target cells, fluorescence labelling with fluorescein-di-beta-Dglucuronide as an enzyme substrate and laser scanning of the membrane filter. Scan results can be confirmed on-line by epifluorescence microscopy. Application to 149 naturally contaminated and uncontaminated well, tap, out-of-pump centre (distribution), surface and sewage spiked water samples indicated > or =90% agreement and equivalence with plate count methods, including Chromocult Coliform agar and m FC agar. In 5.4% of all samples examined, SPC detected between 1 and 11 E. coli per 100 ml, while the two plate methods yielded negative results. Cases of a negative SPC result but a positive E. coli count on both reference media were not observed. This test would primarily be useful for 'emergency' monitoring of drinking water when rapid results are crucial. PMID- 11021571 TI - Antimicrobial mechanism of action of surfactant lipid preparations in enteric Gram-negative bacilli. AB - Two surfactant lipid preparations (SLPs) were investigated to determine their mechanism of antimicrobial action. 8N8, a water-in-oil emulsion, and W60C, a liposome, both have bactericidal activity against Gram-positive bacteria and non enteric Gram-negative bacteria. Additionally, W60C is bactericidal for enteric Gram-negative bacilli when suspended in deionized water. Zeta potential measurements suggested that the resistance of Gram-negative bacilli to 8N8 might be caused by ionic repulsion. Addition of 50 micromol 1(-1) ethylene diamine tetra acetic acid in 100 mmol 1(-1) Tris buffer to either SLPs yielded efficient bactericidal activity against Gram-negative bacilli. This appeared to be due to disruption of the outer membrane and the chelation of divalent cations, as the addition of excess calcium inhibited the antimicrobial effect. Electron microscopy studies documented that 8N8 disrupts the bacterial cell wall, lysing the bacteria, while W60C fuses and internalizes within the cell, causing damage without immediate cell lysis. Understanding the mechanisms of action of these biocidal formulations will help to produce improved formulations with broader spectra of activity. PMID- 11021572 TI - Saccharomyces boulardii stimulates sIgA production and the phagocytic system of gnotobiotic mice. AB - The effect of Saccharomyces boulardii on the immune system was evaluated, comparing germ-free Swiss/NIH mice monoassociated with the probiotic with germ free mice. Saccharomyces boulardii colonized the gut of germ-free mice and survived the gastrointestinal conditions. An increase in sIgA production, both total and anti-S. boulardii, was observed in the intestinal contents of monoassociated mice when compared with germ-free controls. The number of Kupffer cells was significantly higher in monoassociated mice than in germ-free controls. In S. boulardii-monoassociated mice, clearance of Escherichia coli B41 was higher than in germ-free controls. TNF-alpha, IFN-gamma and IL-12 serum levels were higher at earlier time points in monoassociated mice when compared with germ-free mice. These results show that the yeast S. boulardii modulates the host immune responses. This effect may be of interest for improving the resistance to enteropathogenic bacterial infections. PMID- 11021573 TI - The expression of proteinases and haemolysins by Aeromonas hydrophila under modified atmospheres. AB - The study estimated the proteolytic activity (against Hide Powder Azure) and haemolytic activity (against horse erythrocytes) in cell-free filtrates (CFF) from four strains of Aeromonas hydrophila growing under a range of commercially relevant modified atmospheres (2% O2, 78% N2, 20% CO2; 10% O2, 80% N2, 10% CO2; 50% N2, 50% CO2; 100% CO2). The examined strains exhibited significant qualitative and quantitative differences in the extent and times of onset of expression of these enzymes under aerobic and modified atmospheres. No proteolytic or haemolytic activities were detected in any Aer. hydrophila cultures grown at sub-optimal temperatures under modified atmospheres containing high concentrations of CO2 (i.e. 50% CO2 or 100% CO2). Although Aer. hydrophila can grow rapidly in modified atmospheres, the overall spoilage and pathogenic potential is grossly affected. Implications of these findings are discussed. PMID- 11021574 TI - Colonization strategies and conjugal gene transfer of inoculated Pseudomonas syringae on the leaf surface. AB - Survival, colonization and activity of Pseudomonas syringae bacteria inoculated onto the leaf surface of the common bean (Phaseolus vulgaris) was studied. Inoculated Ps. syringae cells shortened by half their size in 100% humidity and by an average of one fifth in 40-60% humidity. The respiring portion of the population, measured by the formation of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC)-formazan crystals, decreased more in 40-60% humidity than in 100% humidity. In scanning electron micrographs, the bacterial cells on leaf surfaces were seen embedded in a mucoid matrix. Intraspecies conjugation of plasmid RP1 also occurred in 40-60% humidity conditions. The portion of transconjugants temporally rose higher than the same portion in 100% humidity conditions. Therefore, although only a small proportion of the inoculated cells remained active on the leaf surface in 40-60% humidity, a relatively high rate of conjugation was still seen. Gene spreading was thus efficient on the leaf surface also when conditions did not allow bacterial population growth. PMID- 11021575 TI - Ultrasonic methodology coupled to ATP bioluminescence for the non-invasive detection of fouling in food processing equipment--validation and application to a dairy factory. AB - The use of an ultrasonic apparatus (40 kHz) for the non-destructive, rapid and reproducible removal of biofilm from standard materials (stainless steel and polypropylene) in a dairy factory was investigated. The application of ultrasound with the tested conditions (10 s and 40 kHz) was found not to be detrimental for standard ATP (concentration ranging between 5 x 10(-9) and 10(-5) mol 1(-1)) and for prokaryotic cells, including both rods and coccoid-shaped bacteria (Escherichia coli and Staphylococcus aureus). It allowed the use of the ATP bioluminescence measurement for quantifying the biofilm removal. The repeatability of industrial milk removal was determined on fouled stainless steel and polypropylene sheets. The variability of the results with the sonication method was constant, +/-24% (coefficient of variation) for both surfaces, and was variable with the swabbing method, +/-42% for the stainless steel sheet and +/ 74% for the polypropylene sheet. The ultrasonic apparatus removed twice the amount of industrial milk biofilm compared with the swabbing method in the case of the polypropylene sheets. The apparatus was used to validate the industrial cleaning protocols of a milk factory. PMID- 11021576 TI - Effects of lactic acid bacteria in inoculants on changes in amino acid composition during ensilage of sterile and non-sterile ryegrass. AB - A study was carried out on the changes occurring in the amino acid fraction of a hybrid ryegrass during ensilage in laboratory-scale silos to help to establish the relative roles of plant and microbial proteases on protein degradation in the silo. Herbage treatments included (i) normal grass without treatment (ii) lambda irradiated grass (sterile) without treatment (iii) sterile, inoculated with a strain of Lactobacillus plantarum and (iv) sterile, inoculated with a strain of Lactobacillus paracasei subsp. paracasei. These treatments had a significant effect on silage amino acid profiles. Concentrations of free amino acids and the extent of amino acid catabolism varied with treatment. However, levels were notably higher in control silages after 90 days (free amino acid nitrogen constituting 54% of total amino acid nitrogen compared with 37, 32 and 22% for treatments i, ii and iv, respectively). These results indicate that the extent of protein hydrolysis during ensilage is influenced by factors other than rate of pH decline and plant protease activity, and that microbial proteases play a role. PMID- 11021577 TI - Comparison of PCR-RFLP, ribotyping and ERIC-PCR for typing Bacillus anthracis and Bacillus cereus strains. AB - PCR-RFLP analysis of the vrrA gene and cerAB gene was used to investigate the genomic diversity in 21 strains of Bacillus anthracis and 28 strains of Bacillus cereus, and was compared with results obtained by ribotyping and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) analysis. VrrA-typing divided the B. anthracis into four groups. Except for one Pasteur vaccine strain, the vrrA PCR-RFLP profiles of the B. anthracis were separated into three groups, which were different from those of the B. cereus strains. Ribotyping separated the B. anthracis isolates into seven ribotypes, and a common fragment of an approximately 850 bp band from the ERIC-PCR fingerprints separated most B. anthracis strains into two groups. VrrA/cerAB PCR-RFLP, ribotyping and ERIC-PCR generated 18, 22 and 23 types, respectively, from B. cereus strains. The results suggest that a combination of all three methods provides a high resolution typing method for B. anthracis and B. cereus. Compared with ribotyping and ERIC-PCR, PCR RFLP is simple to perform and has potential as a rapid method for typing and discriminating B. anthracis strains from other B. cereus group bacteria. PMID- 11021578 TI - Genotypic and phenotypic differentiation of an antifungal biocontrol strain belonging to Bacillus subtilis. AB - Physiological and molecular fingerprints of biotechnologically relevant rhizobacteria are necessary for registration, patenting, recognition and quality checking of the strains. To characterize the biological control agent, Bacillus subtilis B2g, the strain was compared with other plant-associated B. subtilis isolates. Phenotypic characterization included biochemical and nutritional properties, in vitro activity and analysis of potential antagonistic mechanisms towards several plant pathogenic fungi. According to the phenotypic characteristics, it was not possible to differentiate the biocontrol agent from the other strains, although the enzymatic fingerprint was unique. Genotypic diversity among the isolates was characterized by molecular fingerprinting methods using REP-PCR (repetitive extragenomic palindromic PCR), and macrorestriction of genomic DNA and electrophoretic separation of DNA fragments by pulsed-field gel electrophoresis (PFGE). A protocol for PFGE analysis using restriction enzyme SfiI for B. subtilis was developed. PFGE typing of B. subtilis B2g resulted in a unique fingerprint. Therefore, it was possible to differentiate B. subtilis B2g, the biocontrol agent of Phytovit, from other antifungal B. subtilis isolates. PMID- 11021579 TI - Survival of Salmonella in peanut butter and peanut butter spread. AB - In 1996, the first documented outbreak of salmonellosis associated with the consumption of peanut butter was reported. This study was undertaken to determine survival characteristics of high (5.68 log10 cfu g(-1)) and low (1.51 log10 cfu g(-1)) inocula of a five-serotype mixture of Salmonella in five commercial peanut butters and two commercial peanut butter spreads. Populations in samples inoculated with 5.68 log10 cfu g(-1) and stored for 24 weeks at 21 or 5 degrees C decreased 4.14-4.50 log10 cfu g(-1) and 2.86-4.28 log10 cfu g(-1), respectively, depending on the formulation. The order of retention of viability was: peanut butter spreads > traditional (regular) and reduced sugar, low-sodium peanut butters > natural peanut butter. Differences in rates of inactivation are attributed to variation in product composition as well as size and stability of water droplets in the colloidal matrix, which may influence nutrient availability. With the exception of natural peanut butter, products initially inoculated with 1.51 log10 cfu of Salmonella g(-1) (32 cfu g(-1)) were positive for the pathogen after storage for 24 weeks at 5 degrees C. At 21 degrees C, however, with the exception of one peanut butter spread, all products were negative for Salmonella after storage for 24 weeks. Post-process contamination of peanut butter and spreads with Salmonella may to result in survival in these products for the duration of their shelf life at 5 degrees C and possibly 21 degrees C, depending on the formulation. PMID- 11021580 TI - Continuing high prevalence of VanA-type vancomycin-resistant enterococci on Norwegian poultry farms three years after avoparcin was banned. AB - Avoparcin was used as a feed additive in Norwegian broiler and turkey production from 1986 until 1995. It was banned due to the selection of VanA-type vancomycin resistant enterococci (VRE) in animal husbandry and to reduce the potential for human exposure to VRE. The aim of the present study was to investigate the prevalence of VRE carriage in Norwegian poultry farmers and their poultry three years after avoparcin was banned. Corresponding faecal samples from poultry and humans on farms where avoparcin had previously been used (exposed farms, n = 73) and farms where avoparcin had never been used (unexposed farms, n = 74) were analysed for the presence of VRE. For each farm, one sample from the poultry house and one sample from the farmer were obtained. VRE were isolated from 72 of 73 (99%) and eight of 74 (11%) poultry samples from exposed and unexposed farms, respectively. VRE were isolated from 13 of 73 (18%) and one of 74 (1%) farmer samples from exposed and unexposed farms, respectively. All VRE isolates were highly resistant to vancomycin and possessed the vanA gene, as shown by PCR. The high prevalence of VRE is in accordance with previous Norwegian studies, and shows a remarkable stability of the VanA resistance determinant in an apparently non-selective environment. PMID- 11021581 TI - Production of B-group vitamins by two Azotobacter strains with phenolic compounds as sole carbon source under diazotrophic and adiazotrophic conditions. AB - Azotobacter vinelandii strain ATCC 12837 and A. chroococcum strain H23 (CECT 4435) were able to grow on N-free or NH4Cl-amended chemically-defined (Burk's) media, with protocatechuic acid (1-2 mmol 1(-1)) or sodium p-hydroxybenzoate (1 10 mmol 1(-1)) as sole carbon (C) sources. At a concentration of 2 mmol 1(-1), both substrates supported nitrogen fixation (acetylene reduction assay) at similar or higher rates than bacteria grown in control media amended with 2 mmol 1(-1) sodium succinate as C source. The two strains produced the B-group vitamins niacin, pantothenic acid, thiamine, riboflavin and biotin after 72 h of growth in chemically-defined media with 2 mmol 1(-1) protocatechuic acid, sodium phydroxybenzoate or sodium succinate as sole C source, either in N-free media or in media amended with 0.1% NH4Cl. Quantitative production of all vitamins was affected by the use of the different C and N substrates. PMID- 11021582 TI - Occurrence of lectins and hydrophobicity of bacteria obtained from biofilm of hospital catheters and water pipes. AB - Bacteria isolated from biofilms of water distribution pipes and colonized catheters from hospitalized patients were studied for their haemagglutination ability, expression of lectins and hydrophobicity. Higher haemagglutination ability of clinical strains for human red blood cells was demonstrated, which could be an expression of their adaptation to the human ecosystem. Environmental strains had higher hydrophobicity, possibly related to adaptation to a low nutritive ecosystem. Expression of lectins was relatively low and comparable in both bacterial populations, but carbohydrate specificities were very different, possibly related to a different implication of these structures in the two ecosystems. PMID- 11021583 TI - Comparison of two commercial preparations of buffered peptone water for the recovery and growth of Salmonella bacteria from foods. AB - This study compared the performance of two commercial preparations of buffered peptone water. Performance was assessed in terms of ability to resuscitate and recover low numbers of stressed cells, buffering capacity, growth of Salmonella bacteria in pure culture and growth of Salmonella in food pre-enrichments. Although both the preparations of BPW had similar chemical compositions, differences in their recovery performance were found. Brand A recovered significantly higher numbers of heat-injured Salmonella (mean = 0.57 log10 cfu ml(-1) difference) in pure culture compared with brand B when dealing with very low inoculum levels. Although brand B had higher buffering capacity, the pH at the end of the pre-enrichment was found to be similar in both media, even in foods such as milk powder which showed the greatest decline in pH. Both brands were comparable in their ability to grow unstressed Salmonella from different food types. In unstressed cell studies, similar cell numbers were recovered at the end of a 24 h incubation period from both media, although brand B yielded a higher biomass. In the food study with unstressed cells, performance was related more to the food type and the likely association between this and the level and type of competitor organisms present, rather than to the brand of medium used. PMID- 11021584 TI - Use of the DNA sequence of variable regions of the 16S rRNA gene for rapid and accurate identification of bacteria in the Lactobacillus acidophilus complex. AB - The Lactobacillus acidophilus complex includes Lact. acidophilus, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus gasseri and Lactobacillus johnsonii. The objective of this work was to develop a rapid and definitive DNA sequence-based identification system for unknown isolates of the Lact. acidophilus complex. A approximately = 500 bp region of the 16S rRNA gene, which contained the V1 and V2 variable regions, was amplified from the isolates by the polymerase chain reaction. The sequence of this region of the 16S rRNA gene from the type strains of the Lact. acidophilus complex was sufficiently variable to allow for clear differentiation amongst each of the strains. As an initial step in the characterization of potentially probiotic strains, this technique was successfully used to identify a variety of unknown human intestinal isolates. The approach described here represents a rapid and definitive method for the identification of Lact. acidophilus complex members. PMID- 11021585 TI - Bactericidal action of high-power Nd:YAG laser light on Escherichia coli in saline suspension. AB - Infra-red light (1064 nm) from a high-power Nd:YAG laser caused more than 90% loss of viability of Escherichia coli during exposures that raised the temperature of PBS suspensions of the bacteria to 50 C in a thermocouple-equipped cuvette. In contrast, there was minimal loss of viability after heating the same suspensions to 50 degrees C in a water-bath, or in a PCR thermal cycler. The mechanism of laser killing at 50 degrees C was explored by differential scanning calorimetry, by laser treatment of transparent and turbid bacterial suspensions, and by optical absorbency studies of E. coli suspensions at 1064 nm. Taken together, the data suggested that the bactericidal action of Nd:YAG laser light at 50 degrees C was due partly to thermal heating and partly to an additional, as yet undefined, mechanism. Scanning electron microscopy revealed localized areas of surface damage on laser-exposed E. coli cells. PMID- 11021586 TI - Metabolism of arginine and its positive effect on growth and revival of Oenococcus oeni. AB - Oenococcus oeni is the main lactic acid bacteria species which induces malolactic fermentation during wine-making. It is able to break down arginine via the arginine deiminase pathway, a potential source of energy already considered for many bacteria. The production of ATP by starved cells from arginine was quantified with a bioluminescence assay, and efficient coupling of amino acid catabolism and cell growth was monitored. Therefore, molecular growth yield was determined after glucose exhaustion. With colony plate counting and a direct epifluorescence technique, it was shown that addition of arginine to viable but non-culturable cells obtained after nutrient starvation restored their ability to grow during its degradation. Therefore, arginine produced more than maintenance energy. It is concluded that strains which are able to metabolize arginine might take advantage of this additional energy source for growth. PMID- 11021587 TI - Effects of physicochemical variables and cyanobacterial extracts on the immunoassay of microcystin-LR by two ELISA kits. AB - Two types of commercially available ELISA kits for the immunoassay of cyanobacterial microcystins were evaluated for potential interference effects due to methanol, salinity, pH, plasticware and cyanobacterial extract. Of the treatments examined, methanol had the greatest effect, giving false positive microcystin concentrations with increasing methanol concentrations up to 30% (v/v) compared with the negative calibrators of each kit. False positive microcystin results were also produced with increasing salinity up to full strength seawater. Decreases in microcystin-LR equivalents were observed when assaying purified microcystin-LR at pH values between 6.25 and 10. Aqueous microcystin-LR solutions in plastic microcentrifuge tubes after pipetting with disposable plastic tips had lower toxin concentrations than expected when analysed by ELISA. Indicated microcystin concentrations in cyanobacterial extracts varied between kit types and the choice of blanks used. Although ELISAs can be useful tools for the screening of water and cyanobacterial blooms for microcystins and nodularins, users should be aware that commercial kits can be susceptible to interference by commonly encountered environmental and laboratory conditions and materials. PMID- 11021588 TI - Detection of toxigenic Vibrio cholerae from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure. AB - A pit-stop semi-nested PCR assay for the detection of toxigenic Vibrio cholerae in environmental water samples was developed and its performance evaluated. The PCR technique amplifies sequences within the cholera toxin operon specific for toxigenic V. cholerae. The PCR procedure coupled with an enrichment culture detected as few as four V. cholerae organisms in pure culture. Treated sewage, surface, ground and drinking water samples were seeded with V. cholerae and following enrichment, a detection limit of as few as 1 V. cholerae cfu ml(-1) was obtained with amplification reactions from crude bacterial lysates. The proposed method, which includes a combination of enrichment, rapid sample preparation and a pit-stop semi-nested PCR, could be applicable in the rapid detection of toxigenic V. cholerae in environmental water samples. PMID- 11021589 TI - Kinetics of the arginine metabolism of malolactic wine lactic acid bacteria Lactobacillus buchneri CUC-3 and Oenococcus oeni Lo111. AB - The excretion of citrulline, a precursor of carcinogenic ethyl carbamate, formed from arginine degradation by malolactic bacteria in wine is of toxicological concern. The arginine metabolism of resting cells of Lactobacillus buchneri CUC-3 and Oenococcus oeni Lo1l1 was examined. The citrulline excretion rate was found to be linearly correlated to the arginine degradation rate. It was possible to calculate an arginine to citrulline conversion ratio which could be used to predict the amount of citrulline expected after the degradation of a known quantity of arginine. The conversion ratios determined in this study were similar to data calculated from other authors for fermentations in wine and ranged between 4.0% and 7.7%. Ribose, fructose and glucose inhibited the degradation of arginine in Lact. buchneri CUC-3, and inhibition of arginine degradation by glucose correlated with higher arginine to citrulline conversion ratios. The work presents new results of arginine metabolism in malolactic bacteria and gives starting points for investigations in wine. PMID- 11021590 TI - IL-12 plasmid-enhanced DNA vaccination against carcinoembryonic antigen (CEA) studied in immune-gene knockout mice. AB - Intramuscular (i.m.) injection of a plasmid encoding human carcinoembryonic antigen (CEA) elicited immunity against transplanted syngeneic (C57BL/6) CEA positive Lewis lung carcinoma (CEA/LLC) cells, but tumors still appeared in all mice. In wild-type mice, coinjection of an IL-12 plasmid markedly enhanced anti CEA humoral, T-helper-1 and cytotoxic T lymphocyte (CTL) responses, and resistance to a CEA/LLC tumor challenge such that 80% of mice remained tumor free. Injection of the IL- 12 plasmid alone was not protective. To analyze immune requirements, we immunized gene knockout (KO) mice of C57BL/6 background, deficient in either CD3, CD4, CD8, interferon gamma (IFNgamma), perforin or Fas ligand (FasL). Only CD3+ mice expressing both CD4 and CD8, which appear equally important, as well as IFNgamma and perforin, could fully resist a tumor challenge. IL-12 stimulated CTL activity, which was strictly CD3/CD8/perforin dependent. FasL-KO mice had normal CTL activity and tumor resistance, indicating that only the perforin lytic pathway was involved. CD4-KO and IFNgamma-KO mice still generated CTLs. CEA-stimulated IFNgamma production occurred in both CD4- or CD8-KO mice and in both cases was augmented by IL-12. In IFNgamma-KO mice, IL-12 still enhanced anti-CEA antibody production but only moderately restored impaired DTH and tumor resistance. We conclude that the immune requirements for tumor rejection are stringent, involving multiple mechanisms which are all enhanced by IL-12. PMID- 11021591 TI - Transient control of a virus-induced immunopathology by genetic immunosuppression. AB - The ability to control T cell reactivity using suicide genes opens new perspectives for the treatment of T cell-mediated diseases. The therapeutic effect is achieved by the selective killing of thymidine kinase gene-modified activated T cells by ganciclovir (GCV). This strategy has been shown to control T cell alloreactivity efficiently after bone marrow or solid organ transplantation. Here, we aimed to determine whether an immunopathological process induced by a viral infection could be controlled by GCV when T cells express a thymidine kinase transgene. When transgenic mice were infected with the lymphocytic choriomeningitis virus, administration of GCV resulted in an efficient, but only transient, control of the immunopathological immune response. Further analysis revealed the existence of a minute population of GCV-insensitive T cells. These cells expand in response to the virus despite the presence of GCV and cause immunopathology before viral elimination is finally obtained. Thus, when confronted with a replicative virus, the efficacy of this genetic immunosuppression strategy is highly dependent on the presence of even small numbers of GCV-insensitive cells. These results emphasize the need for sufficient preclinical investigations with regard to the pathology and the nature of the immune response if suicide gene transfer is envisioned for new therapeutic indications. PMID- 11021592 TI - Selection of HSV-1 TK gene-transfected murine mammary carcinoma cells resistant to (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) and ganciclovir (GCV). AB - We evaluated the molecular mechanism of resistance in herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene-transfected murine mammary carcinoma (FM3ATK /HSV-1 TK+) cells, that were selected for resistance against (E)-5-(2-bromovinyl) 2'-deoxyuridine (BVDU) and ganciclovir (GCV) by prolonged exposure of the cell cultures to dose-escalating concentrations of these compounds. Drug-resistant FM3ATK-/HSV-1 TK+ cells showed marked differences in their sensitivity spectrum to a series of antiherpetic nucleoside analogues. BVDU-resistant FM3ATK-/HSV-1 TK+ cells displayed the same sensitivity profile as wild-type FM3A/0 cells. In contrast, GCV-resistant FM3ATK-/HSV-1 TK+ cells were still sensitive to BVDU, (E)5-(2-iodovinyl)-2'-deoxyuridine (IVDU) and (E)-5-(2bromovinyl)-2' deoxycytidine (BVDC), a typical feature of FM3A TK cells lacking cytosolic TK. Southern blot and PCR analysis revealed that HSV-1 TK genes were not deleted from the genome of the drug-resistant FM3ATK-/HSV-1 TK+ cells. However, the TK genes in drug-resistant FM3ATK-/HSV-1 TK+ cells were shown to be heavily methylated. Accordingly, RT-PCR demonstrated the complete abrogation of TK mRNA production resulting in a complete loss of TK enzyme activity in drug-resistant FM3A TK-/HSV 1 TK+ cells. PMID- 11021593 TI - Adeno-associated virus vector mediated gene transfer to pancreatic beta cells. AB - Insulin-dependent diabetes mellitus (IDDM) or type 1 diabetes is an autoimmune disease that results in destruction of the insulin-producing pancreatic islet beta cells. Several factors induce the invasion of immune cells into islets and trigger inflammation. Gene therapy approaches targeting the islet cells could be an effective treatment to prevent the onset or reverse type 1 diabetes. Allogeneic islet transplantation provides short-term treatment. However, genetically modified islets, which resist the host immune response, could provide long-term solutions. Adeno-associated virus (AAV) is emerging as a prominent vector system for delivering therapeutic genes for human gene therapy. AAV vector can transduce nondividing cells and provide long-term gene expression by integrating into host chromosome. Therefore, it is an appropriate vector system for islet cell gene therapy. To test the efficacy of AAV vector to transduce pancreatic endocrine cells, we constructed AAV vectors using plasmid pSub201. Wild-type AAV DNA analogue from plasmid psub201 was subcloned into a cloning plasmid pSP72 and AAV vectors were constructed by inserting the transgenes with heterologous promoter in place of AAV open reading frames (rep and cap). In this report we demonstrate the transduction of pancreatic islet cells with AAV vectors encoding bacterial -galactosidase enzyme or enhanced green fluorescent protein (EGFP) as reporter gene. Dispersed porcine and rat islet cells can be transduced by AAV vector, with an efficiency of 47% and 38%, respectively. In particular porcine islet insulin producing beta cells were transduced with an efficiency of 39%. Intact rat islet cells were transduced with an efficiency of 26% as estimated by FACS analysis following transduction with an AAV vector encoding EGFP. Transduction of intact rat islets with an AAV vector did not alter glucose induced insulin secretion. AAV vector transduction was higher in transformed islet cell lines INS-1 and RIN m5F with an efficiency of 65% and 57%, respectively. These new results suggest that AAV vectors will provide an improved method of gene delivery to pancreatic islets and isolated pancreatic beta cells. PMID- 11021594 TI - Cotransduction of nondividing cells using lentiviral vectors. AB - Diseases such as AIDS and cancers may require the introduction of multiple genes into either stem cells or nondividing cells, among others, for therapeutic purposes. Such genes may act at different points of the disease pathway, or may constitute a regulatory loop to bypass or rectify the defective gene or pathway underpinning the disease. Ideally, the therapeutic genes must be transduced together in diverse combinations, and the introduction should occur without constraints. Since lentiviral vectors can transduce both dividing and nondividing cells, they are ideal vehicles to investigate combinatorial gene transfer into diverse cells. In this study, we demonstrate that by using two independent lentiviral vectors, pseudotyped with the protein g of vesicular stomatitis virus, up to four genes can be introduced simultaneously into single dividing and nondividing cells. Up to 45% and 73% of dividing and nondividing cells, respectively, could be transduced with two lentiviral vectors. The efficiency of cotransducing a single cell was the product of the individual transduction efficiencies and suggested the absence of viral interference. Multiple and combinatorial gene transduction using lentiviral vectors may prove useful in gene therapy. PMID- 11021595 TI - Ammonium sulphate precipitation of recombinant adenovirus from culture medium: an easy method to increase the total virus yield. AB - In the majority of the methods for purifying and concentrating recombinant adenoviruses (rAds) the virus that is associated with the helper cells is harvested, while the virus that is present in the cell-culture medium is discarded. During routine propagation of adenovirus type-5 vectors at optimised conditions we noted that, on average, 47% of the total amount of virus is present in the culture medium. To recover and concentrate these rAds from the medium, we devised a method, which is based on ammonium sulphate ((NH4)2SO4) precipitation. At 40% (NH4)2SO4 saturation, 95 +/- 6% of the available virus precipitates from the medium, while the majority of the protein (85%) remains in solution. In contrast to adenovirus precipitation with polyethylene glycol, the (NH4)2SO4 precipitation technique allows collection of precipitated rAds by filtration. We demonstrate here that (NH4)2SO4 precipitation of rAds from cell-culture medium is a simple and fast technique that can be used in combination with standard virus isolation methods to increase the yields of rAds. PMID- 11021596 TI - Poxvirus as a vector to transduce human dendritic cells for immunotherapy: abortive infection but reduced APC function. AB - Dendritic cells (DC) are potent antigen-presenting cells (APC). Ongoing preclinical and clinical studies exploit this capacity for the immunotherapy of tumors. We tested vaccinia virus (VV) as a vector to transduce human DC. Immature and mature DC were prepared from blood monocytes and infected with (1) recombinant VV expressing GFP to analyze infection rates, virus replication in DC and the effect of infection on DC phenotype and (2) recombinant VV expressing beta-galactosidase (betaGAL) under the control of viral early, intermediate and late promoters to analyze the poxvirusdriven gene expression. While the infection rate in DC was comparable to a permissive fibroblast cell line, viral betaGAL gene expression was limited to early promoters. Genes under the control of virus late promoters were not expressed by VV in DC, indicating an abortive infection. VV infection selectively reduced the surface expression of the costimulatory molecule CD80 and the DC maturation marker CD83 on mature DC while other surface molecules including CD86 and MHC remained unchanged. In line with this finding, there was a pronounced reduction in the capacity of VV-infected DC to stimulate allogeneic or autologous T cells in mixed lymphocyte reactions. Furthermore, VV infection inhibited the maturation of immature DC after exposure to proinflammatory cytokines. These results indicate that VV-derived vectors may have complex effects on their target cells. In the case of DC used for immunotherapy, this may be detrimental to their function as potent APC and particularly their capacity to activate T helper cells. PMID- 11021597 TI - Retargeting of adenoviral vectors to neurons using the Hc fragment of tetanus toxin. AB - The Hc fragment of tetanus toxin (Hc) retains the specific nerve cell binding and transport properties of the holotoxin, but lacks any toxicity. We are investigating the potential for utilising its neurotropism for targeted gene delivery to the central nervous system. Previously we reported the use of Hc polylysine conjugates for selective gene transfer into neuronal cells in vitro. However, as attempts to apply these constructs in vivo were not successful, we have extended these studies to modification of the tropism of adenoviral vectors. Either Hc-polylysine conjugates or the Fab fragment of a neutralising anti-knob antibody covalently bound to Hc were attached to the virus. Infection of neuronal and non-neuronal cell lines with retargeted virus showed highly increased neuronal cell selectivity, but no significant enhancement of gene delivery into these cells. High concentrations of free Hc blocked the infectivity of the retargeted vector efficiently. Intramuscular injection of retargeted virus into mouse tongues resulted in selective gene transfer to the neurons of the hypoglossal nucleus, where no pathological changes were observed. As differentiated neurons do not undergo cell division, appropriate vectors carrying a thymidine kinase gene, which allows selective elimination of dividing cells, may be exploitable for the treatment of tumours of the central nervous system. The demonstrated suitability of the Hc fragment of tetanus toxin as targeting moiety for viral vectors also indicates a potential for gene therapy of inherited neurodegenerative diseases such as spinal muscular atrophy. PMID- 11021598 TI - Signaling antibodies complexed with adenovirus circumvent CAR and integrin interactions and improve gene delivery. AB - Current adenoviral (Ad) vectors cannot be targeted to specific cell types due to the widespread distribution of the Ad receptor (CAR). Moreover, CAR and/or internalization receptors (alphav integrins) are absent or present at low levels on some cell types, rendering them resistant to Ad-mediated gene delivery. To address these problems, we have developed a novel vector targeting approach that takes advantage of the common cell signaling pathways initiated by ligation of alphav integrins and growth factor receptors. Recombinant growth factor/cytokines (TNF-alpha, IGF-1, EGF) which trigger phosphatidylinositol-3-OH kinase (PI3K) activation, a signaling molecule involved in adenovirus internalization, were fused to a monoclonal antibody specific for the viral penton base. Ad vectors complexed with these bifunctional mAbs increased gene delivery 10 to 50-fold to human melanoma cells lacking alphav integrins. The bifunctional mAbs also enhanced gene delivery by fiberless adenovirus particles which cannot bind to CAR. Improved gene delivery correlated with increased virus internalization and attachment as well as PI3K activity. The use of bifunctional mAbs to trigger specific cell signaling pathways offers a widely applicable method for bypassing the normal Ad receptors in gene delivery and potentially increasing the selectivity of gene transfer. PMID- 11021599 TI - Stable integration of large (>100 kb) PAC constructs in HaCaT keratinocytes using an integrin-targeting peptide delivery system. AB - Transfer of large DNA constructs in gene therapy studies is being recognised for its importance in maintaining the natural genomic environment of the gene of interest and providing tissue-specific regulation and control. However, methods used to deliver such constructs have been poorly studied. We used a receptor mediated, integrin-targeting transfection system enhanced by liposomes, to deliver a 110 kb PAC (P1-based artificial chromosome) to HaCaT keratinocytes. The PAC contained the collagen VII locus, an EGFP (enhanced green fluorescent protein) reporter gene and the puromycin resistance gene (pac) to allow selection of stably transfected cells. Analysis of puromycin resistant and EGFP-expressing colonies by Western blot showed that collagen VII production increased dramatically after transfection, indicating successful transfer of a large fully functional genomic locus. Fluorescent in situ hybridisation (FISH) and Southern blot analysis revealed that the PAC had integrated as at least one copy per cell. EGFP expression has persisted for 35 weeks, suggesting stable transgene expression. We conclude that the integrin-targeting peptide method of gene delivery is an effective means of stably delivering large DNA constructs to human keratinocytes and could be of benefit for genomic gene therapy approaches. PMID- 11021600 TI - Multiple systemic expression of human interferon-beta in mice can be achieved upon repeated administration of optimized pcTG90-lipoplex. AB - The possibility of achieving multiple systemic expression of human interferon beta in mice upon repeated intravenous administration of cationic liposome-DNA complex (lipoplex) was investigated. Lipoplexes containing the pentammonio lipid pcTG90 were first optimized by selecting the most efficient ratio of pcTG90 to phosphatidylethanolamine (DOPE) and the N/P ratio of cationic lipid nitrogen to DNA phosphate. Highest levels and reproducibility of gene expression were obtained using pcTG90/DOPE (1:2) liposomes complexed with the IFNB1 gene containing plasmid pTG14169 at a N/P ratio of 10. Following lipoplex administration, an early but transient human interferon-beta expression in serum was observed. Importantly, repeated systemic gene expression could be achieved upon re-administration with a minimal time interval of 14 days between two injections. For an interval period of 6 days, subsequent gene expression was inhibited by a first administration of lipoplexes containing either a luciferase reporter gene plasmid or an empty plasmid, but was not inhibited when free (non complexed) plasmid pTG14169 was first injected. Multiple injections of pcTG90 lipoplex performed once every other month resulted in three subsequent peaks of systemic IFNB1 gene expression in mice. In conclusion, our study demonstrates the feasibility of expanding the therapeutic window of a cytokine using repetitive intravenous administration of lipoplex. PMID- 11021601 TI - New leads on ovarian carcinogenesis. PMID- 11021602 TI - Possible health risks of radiofrequency exposure from mobile telephones. PMID- 11021603 TI - Sorting out the role of air pollutants in asthma initiation. PMID- 11021604 TI - Declining ovarian cancer rates in U.S. women in relation to parity and oral contraceptive use. AB - Ovarian cancer incidence and mortality rates have declined among U.S. women age 35-59 years during the period 1970-1995. Epidemiologic studies have shown that ovarian cancer risk decreases with increasing parity and increasing duration of oral contraceptive use. During this period, parity has declined while oral contraceptive use has increased. We compared temporal trends in observed ovarian cancer incidence rates with rates predicted by changes in parity and duration of oral contraceptive use to determine whether the changes in these characteristics could explain the declining rates in younger women. In addition, we wished to examine whether oral contraceptive use continues to be protective to postmenopausal women. To predict changes in rates between 1970 and 1995, we assumed that increases in parity and duration of oral contraceptive use induce proportional decreases in incidence rates. We found that the rates predicted by these assumptions agreed well with observed rates in young women (age 30-49) but were substantially lower than observed rates in older women (age 50-64). The data indicate that the relative decrease in incidence rates due to the protective effect of oral contraceptive use declines with age. PMID- 11021605 TI - Beyond ovulation: oral contraceptives and epithelial ovarian cancer. AB - In a case-control study in three Australian states that included 794 women with epithelial ovarian cancer and 853 community controls for whom we had adequate contraceptive and reproductive histories, we examined the effects of oral contraceptive use after controlling for estimated number of ovulatory cycles. Other covariates included in the multiple logistic regression analysis were parity, smoking, and history of pelvic surgery. The protective effect of duration of oral contraceptive use appeared to be multiplicative, with a 7% decrease in relative risk per year [95% confidence interval (CI) = 4-9%], persisting beyond 15 years of exposure. Use for up to 1 year may have a greater effect than predicted (odds ratio = 0.57; 95% CI = 0.40-0.82), whereas use before the first pregnancy may be additionally beneficial (odds ratio = 0.95; 95% CI = 0.87-1.03, adjusted for overall duration of use). Better control for ovulatory life might attenuate these estimates somewhat. There was little evidence of waning protection with time since last exposure or of extra benefit with early commencement of oral contraceptive use. We found no convincing evidence of effect modification in any factor examined or differences in effect among the three main histologic cancer types or between borderline and malignant tumors. Oral contraceptives may act by both suppressing ovulation and altering the tumor promoting milieu. PMID- 11021606 TI - Factors related to inflammation of the ovarian epithelium and risk of ovarian cancer. AB - Previous epidemiologic observations consistently suggest that suppression of ovulation, tubal ligation, and hysterectomy reduce the risk of ovarian cancer and that perineal talc use increases the risk. We examined these and other risk factors in the context of a new hypothesis: that inflammation may play a role in ovarian cancer risk. Ovulation entails ovarian epithelial inflammation; talc, endometriosis, cysts, and hyperthyroidism may be associated with inflammatory responses of the ovarian epithelium; gynecologic surgery may preclude irritants from reaching the ovaries via ascension from the lower genital tract. We evaluated these risk factors in a population-based case-control study. Cases 20 69 years of age with a recent diagnosis of epithelial ovarian cancer (767) were compared with community controls (1,367). We found that a number of reproductive and contraceptive factors that suppress ovulation, including gravidity, breast feeding, and oral contraception, reduced the risk of ovarian cancer. Environmental factors and medical conditions that increased risk included talc use, endometriosis, ovarian cysts, and hyperthyroidism. Gynecologic surgery including hysterectomy and tubal ligation were protective. Tubal ligation afforded a risk reduction even 20 or more years after the surgery. The spectrum of associations provides support for the hypothesis that inflammation may mediate ovarian cancer risk. PMID- 11021607 TI - Radiofrequency exposure and mortality from cancer of the brain and lymphatic/hematopoietic systems. AB - The proliferation of wireless communication technologies has raised public concern regarding potential health effects of radiofrequency (RF) exposures. This is the first report of findings from a large-cohort mortality study among employees of Motorola, a manufacturer of wireless communication products. We examined all major causes of mortality, with brain cancers, lymphomas, and leukemias as a priori outcomes of interest. Using job titles, we classified workers into high, moderate, low, and background RF exposure groups. A total of 195,775 workers contributed 2.7 million person-years during the 1976-1996 period. Using external comparisons, the standardized mortality ratios for RF-exposed workers were 0.53 [95% confidence interval (CI) = 0.21-1.09] and 0.54 (95% CI = 0.33-0.83) for central nervous system/brain cancers and all lymphomas/leukemias. Rate ratios calculated from Poisson regression models based on internal comparisons were near 1.0 for brain cancers and below 1.0 for all lymphomas and leukemias. These findings were consistent across cumulative, peak, and usual exposure classifications. We did not observe higher risk with increased exposure duration or latency. Although this study is limited by the use of a qualitative exposure matrix and the relatively young age of the cohort, our findings do not support an association between occupational RF exposure and brain cancers or lymphoma/leukemia. PMID- 11021608 TI - The relation between infant indoor environment and subsequent asthma. AB - Our aim was to examine the contribution of an infant's indoor environment to childhood asthma using prospective data. We conducted a cross-sectional asthma survey in 1995 on 92% (6,378/6,911) of 7-year-olds in Tasmania, Australia. We linked these data with data collected in 1988 as part of the Tasmanian Infant Health Survey, which was designed to investigate sudden infant death. We were able to match 863 records out of the 1,111 in the 1988 survey and the 6,378 in the 1995 survey. The former group was interviewed at home at 1 month of age. In homes where at least one adult smoked in 1988, reported infant exposure to smoking in the same room in 1988 was associated with increased asthma by 1995 (relative risk = 1.52; 95% confidence interval = 1.01-2.29) after adjustment for confounders. The associations between infant exposure to environmental tobacco smoke and asthma were not consistent, however. Gas heater use in 1988 was associated with asthma (relative risk = 1.92; 95% confidence interval = 1.33 2.76). Markers of aeroallergen exposure at 1 month of age were not materially associated with asthma or wheeze. In some settings, air circulation practice with regard to bedroom door closure appeared important. Poor indoor air quality may play an important role in the development of childhood asthma. PMID- 11021609 TI - Associations between outdoor air pollutants and hospitalization for respiratory diseases. AB - The concentration of particulate matter in outdoor air, as indicated by daily measures of particulate matter 10 microm in diameter (PM10) in many cities, has been found to be associated with the daily number of deaths and hospital admissions in these cities. To understand this association better, we studied the daily number of hospital admissions for respiratory diseases and the concentrations of eight pollutants in ambient air, during a period of 38 months, in an area with cold winters and air pollution that comes mainly from motor vehicles. We estimated the changes in risk of hospitalization by interquartile increases in pollutant concentrations by Poisson regression analyses. Controlling for periodic trends and weather, the relative risk of hospitalization associated with an interquartile increase of PM10 was 1.038 [95% confidence interval (CI) = 0.991-1.087]. In contrast, the relative risk associated with benzene was 1.105 (95% CI = 1.047-1.166). In a two-pollutant model, the relative risk estimates were 1.014 (95% CI = 0.966-1.063) for PM10 and 1.090 (95% CI = 1.031-1.153) for benzene. We evaluated other two- and three-pollutant models and concluded that pollutants other than PM10 are more strongly associated with hospital admissions for respiratory diseases. PMID- 11021610 TI - Time to pregnancy among the wives of men occupationally exposed to lead. AB - We conducted a retrospective study on time to pregnancy among the wives of men biologically monitored for lead to assess whether paternal occupational exposure to inorganic lead is associated with decreased fertility. After three mailings, 554 (72.2%) wives of the monitored men participated. The final study population consisted of 502 couples who did not use contraception at the beginning of the pregnancy. We assessed lead exposure from blood lead measurements and from questionnaires completed by the men. We calculated the relative fecundability density ratios with discrete proportional hazards regression. The fecundability density ratios, adjusted for potential confounders, were 0.92 [95% confidence interval (95% CI) = 0.73-1.16], 0.89 (95% CI = 0.66-1.20), 0.58 (95% CI = 0.33 0.96), and 0.83 (95% CI = 0.50-1.32) for blood lead categories 0.5-0.9, 1.0-1.4, 1.5-1.8, and > or =1.9 micromol/L, respectively, as compared with <0.5 micromol/L. The findings provide limited support for the hypothesis that paternal exposure to lead is associated with decreased fertility. PMID- 11021611 TI - Paternal exposure to lead and infertility. AB - To assess whether paternal exposure to lead is associated with infertility, we performed a register-based study among married men biologically monitored for exposure to inorganic lead. We obtained information about the marriages and the wives of the men from the Finnish Central Population Register. Data on pregnancies were obtained from medical records. Paternal exposure to lead was assessed on the basis of blood lead measurements. We estimated the risk of infertility, defined as nonoccurrence of a marital pregnancy, by applying binomial regression. For the blood lead categories of 0.5-0.9, 1.0-1.4, 1.5-1.9, 2.0-2.4, and > or =2.5 micromol/L the relative risk of infertility, compared with the risk in the lowest exposure category (<0.5 micromol/L), was 1.27 (95% confidence interval 1.08-1.51), 1.35 (1.12-1.63), 1.37 (1.08-1.72), 1.50 (1.08 2.02), and 1.90 (1.30-2.59), respectively. The findings support the hypothesis that paternal exposure to lead increases the risk of infertility at low occupational exposure levels. We applied proportional hazards regression to the analysis of pregnancy delay. A delay was observed among the wives of men exposed to lead. Exposure to lead was not clearly associated with delayed pregnancy, however, when the analysis was restricted to couples with at least one pregnancy. This finding suggests that the restriction of the study on time to pregnancy to fertile couples may introduce a bias toward no association. PMID- 11021612 TI - Outdoor air concentrations of nitrogen dioxide and sulfur dioxide and prevalence of wheezing in school children. AB - We report analysis of data on outdoor air pollution and respiratory symptoms in children collected in the Czech part of the international Small Area Variations in Air pollution and Health (SAVIAH) Project, a methodological study designed to test the use of geographical information systems (GIS) in studies of environmental exposures and health at small area level. We collected the following data in two districts of Prague: (1) individual data on 3,680 children (response rate 88%) by questionnaires; (2) census-based socio-demographic data for small geographical units; (3) concentrations of nitrogen dioxide (NO2) and sulfur dioxide (SO2) measured by passive samplers in three 2-week surveys at 80 and 50 locations, respectively. We integrated all data into a geographical information system. Modeling of NO2 and SO2 allowed estimation of exposure to outdoor NO2 and SO2 at school and at home for each child. We examined the associations between air pollution and prevalence of wheezing or whistling in the chest in the last 12 months by logistic regression at individual level, weighted least squares regression at small area (ecological) level and multilevel modeling. The results varied by the level of analysis and method of exposure estimation. In multilevel analyses using individual data, odds ratios per 10 microg/m3 increase in concentrations were 1.16 (95% CI = 0.95-1.42) for NO2, and 1.08 (95% CI = 0.97-1.21) for SO2. While mapping of spatial distribution of NO2 and SO2 in the study area appeared valid, the interpolation from outdoor to personal exposures requires consideration. PMID- 11021613 TI - Pregnancy outcomes in females after treatment for childhood cancer. AB - This Ontario province-wide cohort study was conducted to compare the risk of adverse pregnancy outcomes in female childhood cancer survivors who received abdominal-pelvic radiation and/or chemotherapy with alkylating agents with the risk among those who were treated by non-sterilizing alkylating agents with the risk among those who were treated by non-sterilizing surgery only. Females in Ontario, Canada, diagnosed in 1964-1988 before age 20 with a histologically confirmed malignancy and who had survived for at least 5 years, attained age 18, and were alive at the time of study, were identified through the Ontario Cancer Registry. We ascertained pregnancy outcomes by a telephone-administered questionnaire. Treatment data were abstracted from medical records for 830 subjects 18-49 years of age, the analysis comprised 340 survivors who had one or more pregnancies after treatment. There was no evidence of an increased risk of having a spontaneous abortion or an infant with a birth defect. Survivors receiving abdominal-pelvic radiation were more likely to have a low birth weight infant (odds ratio estimate [OR] = 3.64; 95% confidence interval [CI] = 1.33 9.96), a premature low birth weight infant (OR = 3.29; 95% CI = 0.97-11.1), or an infant who died in the perinatal period (OR = 2.41; 95% CI = 0.50-11.5), compared with those receiving surgery. Risks of perinatal death and having a low birth weight infant increased with dose of radiotherapy directed to the abdomen. PMID- 11021614 TI - Implications of stage-specific survival rates in assessing recent declines in prostate cancer mortality rates. AB - It has been noted that the most important evidence for a benefit of early detection of prostate cancer using prostate-specific antigen (PSA) testing would be a decline in prostate cancer mortality rates to levels below those existing before diagnostic use of PSA testing. We document a decrease in U.S. prostate cancer mortality rates in white men less than 85 years of age to levels below those existing in 1986, the year use of PSA testing was approved. In fact, for men 60-79 years of age, prostate cancer mortality rates were lower in 1997 than in any year since 1950. Although it has been argued that the decrease in prostate cancer mortality rates began too soon to be explained by PSA testing, stage specific survival rates indicate that a rapid decrease in mortality may be explained by the large number of high-grade prostate cancers detected before metastasis. If recent decreases in U.S. prostate cancer mortality rates are due to early detection using PSA testing, randomized clinical trials investigating PSA testing will show early evidence of a mortality benefit. PMID- 11021615 TI - Antidepressant medications and risk for cancer. AB - Antidepressants appear to promote tumor growth in experimental studies; however, results from epidemiologic studies are inconclusive. We used a population-based cohort study to estimate the incidence of cancer after antidepressant treatment in 39,807 adult users of antidepressants identified in the Prescription Database of the County of North Jutland, Denmark between January 1, 1989 and December 31, 1995. Information on cancer occurrence was obtained from the Danish Cancer Registry. We categorized exposure according to use of tricyclic antidepressants, tetracyclic antidepressants, selective serotonin reuptake inhibitors, or monoamine oxidase inhibitors. In the follow-up period beginning 1 year after first known prescription, there were 966 cancers among users of antidepressants; our population estimate suggested an expected number of 946 for an overall standardized incidence ratio of 1.0 (95% confidence interval = 1.0-1.1). Users of tricyclic antidepressants had an excess of non-Hodgkin's lymphoma, with the risk increasing with the number of prescriptions of tricyclic antidepressants. The standardized incidence ratio was 2.5 (95% confidence interval, 1.4-4.2) for those with five or more prescriptions. Our results provide little evidence that antidepressants promote cancer at other sites, except for a possible effect of tricyclic antidepressants and tetracyclic antidepressants on non-Hodgkin's lymphoma. PMID- 11021616 TI - Induced abortion in relation to breast cancer among parous women: a birth certificate registry study. AB - We wished to assess the relation of induced abortion to the subsequent incidence of breast cancer among parous women, using a design that would prevent the possibility of differentially complete reporting of abortion history by women with breast cancer and controls. Our study was conducted within a cohort of women who gave birth to a child during 1984-1994 while residing in 13 counties of western Washington. Cases were women from the cohort diagnosed with breast cancer between 1984 and 1994. From the remaining cohort members, five controls were matched to each woman with breast cancer by year of index birth (ie, the last child born before breast cancer diagnosis) and by age at delivery. We categorized 463 cases and 2,201 controls according to history of induced abortion as recorded on the index birth certificate. The risk of breast cancer was not found to be associated with a prior induced abortion (estimated relative risk (RR) = 0.9, 95% confidence interval (CI) 0.7-1.2). These results suggest that an induced abortion, if followed at some later time by pregnancy and childbirth, does not increase a woman's risk of breast cancer. PMID- 11021617 TI - Left-handedness in relation to breast cancer risk in postmenopausal women. AB - Breast cancer risk may be influenced by intrauterine exposure to steroid hormones. We evaluated left-handedness, a marker of intrauterine hormone exposure, in relation to breast cancer risk in our population-based, case-control study. Case women 50-79 years of age with a first diagnosis of invasive breast cancer were ascertained through statewide cancer registries in Wisconsin, Massachusetts, and New Hampshire. Control women were identified in each state through lists of licensed drivers (for ages 50-64) and Medicare beneficiaries (for ages 65-79), and selected at random to correspond with the age distribution of case women. Exposure information, including handedness, was obtained through a telephone interview. Our results indicated a modest association between left handedness and breast cancer risk (OR = 1.42; 95% CI = 1.10-1.83). The effect of left-handedness was modified by age; we observed the greatest risk ratio in the oldest age group. Left-handedness was not associated with breast tumor laterality. Our results are consistent with the hypothesis that intrauterine hormone exposures play a role in the development of breast cancer. PMID- 11021618 TI - Low birth weight and preterm delivery as risk factors for asthma and atopic dermatitis in young adult males. AB - Gestational factors have been hypothesized to play a role in the susceptibility to asthma and atopic dermatitis. We examined whether fetal growth was associated with asthma and atopic dermatitis separately in a population of 4,795 male conscripts born between 1973 and 1975 in Denmark. The prevalence of asthma was 4.7%. The prevalence odds ratio of asthma in conscripts with a birth weight below 2,501 g was 1.5 (95% confidence interval = 0.7-3.1) compared with conscripts with a birth weight of 3,001-3,500 g, adjusted for gestational age and potential confounders. The adjusted prevalence odds ratio among conscripts born before 34 gestational weeks was 0.8 (95% confidence interval = 0.3-2.0) compared with conscripts born at term. The prevalence of atopic dermatitis was 1.0%. The prevalence odds ratio of atopic dermatitis among those with a birth weight below 2,501 g was 3.0 (95% confidence interval = 0.8-11.9) compared with those whose birth weight was between 3,001 and 3,500 g. Men whose gestational age had been below 34 weeks had an adjusted prevalence odds ratio of 0.3 (95% confidence interval = 0.0-3.1). These findings indicate that fetal growth retardation rather than preterm delivery of male infants is the main gestational factor underlying the associations but does not explain the apparent increase over time of asthma or atopic diseases. PMID- 11021619 TI - Do confounding or selection factors of residential wiring codes and magnetic fields distort findings of electromagnetic fields studies? AB - In contrast with several previous studies, our recent large case-control study found little association between childhood acute lymphoblastic leukemia (ALL) and electric-power-line wire codes. Here we examine internal evidence from our study to assess the possibility that selection bias and/or confounding may have affected the findings. We compared the relation between childhood ALL and wire codes and direct measurements of magnetic fields in subjects who participated in all phases of the study with the relation in all subjects, including those who declined to allow access inside the home. We found that the odds ratio for ALL among those living in homes with very high current configurations increased by 23% when 107 "partial participants" were excluded. We found similar, but slightly smaller, increases in the odds ratios when we performed the same comparisons using direct measurements of magnetic fields, excluding subjects who allowed only a measurement outside the front door. "Partial participants" tended to be characterized by lower socioeconomic status than subjects who participated fully, suggesting possible selection bias. We also examined the relation between a large number of potential confounding variables and both proxy and direct measurements of magnetic fields. Univariate adjustment for individual variables changed the odds ratio for ALL by less than 8%, while simultaneous adjustment for several factors reduced the estimate by a maximum of 15%. We conclude that while confounding alone is unlikely to be an important source of bias in our own and previous studies of magnetic fields, selection bias may be more of a concern, particularly in light of the generally low response rates among controls in case control studies. PMID- 11021620 TI - An exploration of awareness bias in two environmental epidemiology studies. AB - Awareness bias in environmental health research is the tendency to report more illness because of concerns arising from proximity to a hazard in the absence of a measurable biological effect. Such bias complicates the interpretation of self reported symptoms in communities exposed to emissions from heavy industry. We used data from two epidemiologic studies in Northeast England where community concerns existed about health risks from industry. An association between proximity to industry and self-reported respiratory and nonrespiratory illnesses and symptoms had been found in one study but not in the other. An indicator of concern about industrial pollution was constructed from responses to a 17-item questionnaire about issues that had caused stress or anxiety. Univariate and multivariate analyses of health outcome variables in both studies showed that individuals with "industry-related worries" reported more illness, irrespective of proximity to industry. We conclude that self-reported illness was influenced by both worry and proximity to industry, but that worry about the hazard had the greatest effect on self-reported illness. We suggest that because absolute certainty about the role and extent of awareness bias in environmental epidemiology studies is unlikely to be achieved, self-reported data should be supplemented with other observations. PMID- 11021621 TI - Estimation of completeness of AIDS case reporting in Massachusetts. AB - One of the most important aspects of any surveillance system is degree of completeness. We conducted a multiple source capture-recapture study using the 1994 Massachusetts Uniform Hospital Discharge Data Set (UHDDS) and Medicaid claims data to evaluate the completeness of the state's AIDS registry. We used encrypted social security numbers as the primary link to ensure confidentiality. For cases that did not link in the first round owing to missing social security numbers, we linked using gender and date of birth. Staff reviewed unmatched records from the Uniform Hospital Discharge Data Set and Medicaid datasets to determine subjects' AIDS case status. Using the Uniform Hospital Discharge Data Set, the AIDS registry was 92.6% complete (95% confidence interval (CI) = 91.6 93.5). The Medicaid claims dataset suggested the AIDS registry to be 94.5% complete (95% confidence interval = 93.7-95.3). The completeness of reporting to the state AIDS registry continues to be high, but there are differences by gender and mode of transmission of the virus. The continued assessment of completeness of reporting is important to ensure quality of the surveillance database over time. PMID- 11021622 TI - Body size and hip fracture risk. Swedish Hip Fracture Study Group. AB - The objective of this population-based case-control study was to determine the independent association between height, weight at different ages and adult weight change on hip fracture risk, and the joint effects of these factors. The study base comprised postmenopausal women 50-81 years of age who resided in six counties in Sweden during the period October 1993 to February 1995. The study included 1,327 cases with an incident hip fracture and 3,262 randomly selected controls. We obtained information on body measures and other factors possibly related to hip fracture through mailed questionnaires and telephone interviews. Height and weight change were dominant risk factors. Tall women (> or = 169 cm) had an odds ratio of 3.16 (95% confidence interval = 2.47-4.05) compared with women shorter than 159 cm. Weight gain during adult life was strongly protective: compared with those with moderate weight change (-3 to 3 kg), those with substantial weight gain (> or =12 kg) had a markedly decreased risk of hip fracture (odds ratio = 0.35; 95% confidence interval = 0.27-0.45), whereas weight loss was associated with an increased risk. Weight change retained important effects among all subjects, even after controlling for current weight and weight at age 18. In contrast, among women who gained weight, the separate effects of current weight and weight at age 18 were small or absent. Among women who lost weight, both current weight and weight at age 18 had effects that remained after controlling for weight change. Adult weight change and height are dominant body size risk factors for hip fracture. Weight loss vs weight changes demarcates different patterns of hip fracture risk. PMID- 11021624 TI - Ethical considerations concerning the regulation of human exposure to electromagnetic fields. PMID- 11021623 TI - Epstein-Barr virus and multiple sclerosis. AB - It has been suggested that the Epstein-Barr virus (EBV) plays a role in the etiology of multiple sclerosis (MS), but individual epidemiologic studies have been inconclusive, in part because of the high prevalence of previous infection among individuals without MS. We conducted a systematic review of case-control studies comparing EBV serology in MS patients and controls. Eight published investigations were identified, including a total of 1,005 cases and 1,060 controls. The summary odds ratio of MS comparing EBV seropositive individuals with EBV seronegative individuals was 13.5 (95% CI = 6.3-31.4). The strength and consistency of this association and the high sensitivity and specificity of EBV serology suggest that these results are not readily explained by an aspecific immune activation among MS patients. These findings support a role of EBV in the etiology of MS. PMID- 11021625 TI - Does randomization introduce bias in unblinded trials? PMID- 11021626 TI - Trends of spontaneous abortions in Italy 1990-1995. PMID- 11021627 TI - Maternal pesticide exposure and childhood leukemia. PMID- 11021628 TI - Parental smoking and infection with Helicobacter pylori among preschool children. PMID- 11021629 TI - Serum levels of testosterone do not provide evidence of selection bias in studies of male reproductive health. PMID- 11021630 TI - Induced abortion and risk of breast cancer. PMID- 11021631 TI - Osteopontin. AB - Osteopontin (OPN) is a highly phosphorylated sialoprotein that is a prominent component of the mineralized extracellular matrices of bones and teeth. OPN is characterized by the presence of a polyaspartic acid sequence and sites of Ser/Thr phosphorylation that mediate hydroxyapatite binding, and a highly conserved RGD motif that mediates cell attachment/signaling. Expression of OPN in a variety of tissues indicates a multiplicity of functions that involve one or more of these conserved motifs. While the lack of a clear phenotype in OPN "knockout" mice has not established a definitive role for OPN in any tissue, recent studies have provided some novel and intriguing insights into the versatility of this enigmatic protein in diverse biological events, including developmental processes, wound healing, immunological responses, tumorigenesis, bone resorption, and calcification. The ability of OPN to stimulate cell activity through multiple receptors linked to several interactive signaling pathways can account for much of the functional diversity. In this review, we discuss the structural features of OPN that relate to its function in the formation, remodeling, and maintenance of bones and teeth. PMID- 11021632 TI - Genetic and teratogenic approaches to craniofacial development. AB - Craniofacial malformations are the most common birth defects that occur in humans, with facial clefting representing the majority of these defects. Facial clefts can arise at any stage of development due to perturbations that alter the extracellular matrix as well as affect the patterning, migration, proliferation, and differentiation of cells. In this review, we focus on recent advances in the understanding of the developmental basis for facial clefting through the analysis of the effects of gene disruption experiments and treatments with teratogens in both chickens and mice. Specifically, we analyze the results of disruptions to genes such as Sonic hedgehog (Shh), epidermal growth factor receptor (EGFR), Distal-less (Dlx), and transforming growth factor beta 3 (TGFbeta3). We also describe the effects that teratogens such as retinoic acid, jervine, and cyclopamine have on facial clefting and discuss mechanisms for their action. In addition to providing insight into the bases for abnormal craniofacial growth, genetic and teratogenic techniques are powerful tools for understanding the normal developmental processes that generate and pattern the face. PMID- 11021633 TI - Molecular signaling and pulpal nerve development. AB - The purpose of this review is to discuss molecular factors influencing nerve growth to teeth. The establishment of a sensory pulpal innervation occurs concurrently with tooth development. Epithelial/mesenchymal interactions initiate the tooth primordium and change it into a complex organ. The initial events seem to be controlled by the epithelium, and subsequently, the mesenchyme acquires odontogenic properties. As yet, no single initiating epithelial or mesenchymal factor has been identified. Axons reach the jaws before tooth formation and form terminals near odontogenic sites. In some species, local axons have an initiating function in odontogenesis, but it is not known if this is also the case with mammals. In diphyodont mammals, the primary dentition is replaced by a permanent dentition, which involves a profound remodeling of terminal pulpal axons. The molecular signals underlying this remodeling remain unknown. Due to the senescent deterioration of the dentition, the target area of tooth nerves shrinks with age, and these nerves show marked pathological-like changes. Nerve growth factor and possibly also brain-derived neurotrophic factor seem to be important in the formation of a sensory pulpal innervation. Neurotrophin-3 and -4/5 are probably not involved. In addition, glial cell line-derived neurotrophic factor, but not neurturin, seems to be involved in the control of pulpal axon growth. A variety of other growth factors may also influence developing tooth nerves. Many major extracellular matrix molecules, which can influence growing axons, are present in developing teeth. It is likely that these molecules influence the growing pulpal axons. PMID- 11021634 TI - Biocompatibility of resin-modified filling materials. AB - Increasing numbers of resin-based dental restorations have been placed over the past decade. During this same period, the public interest in the local and especially systemic adverse effects caused by dental materials has increased significantly. It has been found that each resin-based material releases several components into the oral environment. In particular, the comonomer, triethyleneglycol di-methacrylate (TEGDMA), and the 'hydrophilic' monomer, 2 hydroxy-ethyl-methacrylate (HEMA), are leached out from various composite resins and 'adhesive' materials (e.g., resin-modified glass-ionomer cements [GICs] and dentin adhesives) in considerable amounts during the first 24 hours after polymerization. Numerous unbound resin components may leach into saliva during the initial phase after polymerization, and later, due to degradation or erosion of the resinous restoration. Those substances may be systemically distributed and could potentially cause adverse systemic effects in patients. In addition, absorption of organic substances from unpolymerized material, through unprotected skin, due to manual contact may pose a special risk for dental personnel. This is borne out by the increasing numbers of dental nurses, technicians, and dentists who present with allergic reactions to one or more resin components, like HEMA, glutaraldehyde, ethyleneglycol di-methacrylate (EGDMA), and dibenzoyl peroxide (DPO). However, it must be emphasized that, except for conventional composite resins, data reported on the release of substances from resin-based materials are scarce. There is very little reliable information with respect to the biological interactions between resin components and various tissues. Those interactions may be either protective, like absorption to dentin, or detrimental, e.g., inflammatory reactions of soft tissues. Microbial effects have also been observed which may contribute indirectly to caries and irritation of the pulp. Therefore, it is critical, both for our patients and for the profession, that the biological effects of resin-based filling materials be clarified in the near future. PMID- 11021635 TI - Smoking and periodontal disease. AB - Numerous investigations of the relationship between smoking and periodontal disease have been performed over the last 15 years, and there now exists a substantial body of literature upon which this current review is based. From both cross-sectional and longitudinal studies, there appears to be strong epidemiological evidence that smoking confers a considerably increased risk of periodontal disease. This evidence is further supported by the data emanating from patients who stop smoking. These patients have levels of risk similar to those of non-smokers. Numerous studies of the potential mechanisms whereby smoking tobacco may predispose to periodontal disease have been conducted, and it appears that smoking may affect the vasculature, the humoral immune system, and the cellular immune and inflammatory systems, and have effects throughout the cytokine and adhesion molecule network. The aim of this review is to consider the evidence for the association between smoking and periodontal diseases and to highlight the biological mechanisms whereby smoking may affect the periodontium. PMID- 11021636 TI - Food starches and dental caries. AB - Sucrose and starches are the predominant dietary carbohydrates in modern societies. While the causal relationship between sucrose and dental caries development is indisputable, the relationship between food starch and dental caries continues to be debated and is the topic of this review. The current view of dental caries etiology suggests that in-depth evaluation of the starch-caries relationship requires the consideration of several critical cariogenic determinants: (1) the intensity (i.e., the amount and frequency) of exposure of tooth surfaces to both sugars and starches, (2) the bioavailability of the starches, (3) the nature of the microbial flora of dental plaque, (4) the pH lowering capacity of dental plaque, and (5) the flow rate of saliva. Studies of caries in animals, human plaque pH response, and enamel/dentin demineralization leave no doubt that processed food starches in modern human diets possess a significant cariogenic potential. However, the available studies with humans do not provide unequivocal data on their actual cariogenicity. In this regard, we found it helpful to distinguish between two types of situations. The first, exemplified by our forebears, people in developing countries, and special subject groups in more modern countries, is characterized by starch consumption in combination with a low sugar intake, an eating frequency which is essentially limited to two or three meals per day, and a low-to-negligible caries activity. The second, exemplified by people in the more modern societies, e.g., urban populations, is characterized by starch consumption in combination with significantly increased sugar consumption, an eating frequency of three or more times per day, and a significantly elevated caries activity. It is in the first situation that food starches do not appear to be particularly caries-inducive. However, their contribution to caries development in the second situation is uncertain and requires further clarification. Although food starches do not appear to be particularly caries inducive in the first situation, the possibility cannot be excluded that they contribute significantly to caries activity in modern human populations. The commonly used term "dietary starch content" is misleading, since it represents a large array of single manufactured and processed foods of widely varying composition and potential cariogenicity. Hence, increased focus on the cariogenicity of single starchy foods is warranted. Other aspects of starchy foods consumption, deserving greater attention, include the bioavailability of starches in processed foods, their retentive properties, also in relation to sugars present (starches as co-cariogens), their consumption frequency, the effect of hyposalivation on their cariogenicity, and their impact on root caries. The starch-caries issue is a very complex problem, and much remains uncertain. More focused studies are needed. At present, it appears premature to consider or promote food starches in modern diets as safe for teeth. PMID- 11021638 TI - Histone deacetylase inhibitors and cell proliferation in pea root meristems. AB - The histone deacetylase (HDA) inhibitors, trichostatin A (TSA) and HC toxin halt mitosis in cultured root meristems of Pisnum sativum, while the anti-protozoan HDA inhibitor apicidin is ineffective. Two-dimensional PAGE of proteins from root meristems exposed to TSA and HC toxin did not show significant differences compared to controls, although a previously tested HDA inhibitor, butyrate, exhibited dramatic variations in its protein profile. Northern analysis of butyrate- and TSA-treated root meristems indicated that non-proliferating cells are expressing significant amounts of transcripts of the known cell proliferation associated genes: histone H2A, MAP kinase, cycA2:1 and cdc2. Western analysis reveals the presence of hyperacetylated nuclear proteins in HDA-inhibitor treated cells. These results suggest that the HDA inhibitors, butyrate and TSA, halt mitosis without down-regulating genes that typically have low or nonexistent expression levels in non-dividing cells. PMID- 11021637 TI - Gibbestatin B inhibits the GA-induced expression of alpha-amylase expression in cereal seeds. AB - The expression of alpha-amylase in aleurone layers of barley is known to be induced by gibberellin A3 (GA). In the present study, gibbestatin B (GNB) was isolated from Streptomyces sp. C-39 as an inhibitor of the GA-induced expression of alpha-amylase in barley and rice, with IC50 values of 125 and 70 microM, respectively. GNB suppressed accumulation of GA-induced barley high-pI type B and rice RAmylA alpha-amylase transcripts. However, GNB showed no inhibitory activity on GUS expression in transgenic tobacco harboring the auxin-inducible par B promoter:: GUS fusion gene. The transcription of an abscisic acid (ABA)-inducible gene, HVA1, was unaffected by GNB. In addition, GNB prevented aleurone cells from cell death induced by GA. In tobacco and Arabidopsis plants, GNB suppressed the germination and retarded the growth of seedlings without toxicity. The growth of gai, spy and abi mutants was also retarded by GNB. Normal plants treated with GA biosynthesis inhibitors and GA-defective and GA-signaling mutants normally have dwarf dark green leaves. However, dwarfed healthy green leaves were observed in normal plants treated with GNB. GA-induced stem elongation of plants was also detected in the presence of GNB. These analyses indicate that GNB inhibits the GA induced expression of alpha-amylase by regulating one of the steps involved in ABA signaling, but not by acting as a weak ABA analog. PMID- 11021639 TI - Transformation of jervine by Cunninghamella elegans ATCC 9245. AB - Preparative-scale fermentation of the known C-nor-D-homosteroidal jerveratrum alkaloid jervine with Cunninghamella elegans (ATCC 9245) has resulted in the isolation of (-)-jervinone as the major metabolite. In addition, C. elegans ATCC 9245 was able to epimerize C-3 of jervine, producing 3-epi-jervine. This epimerization reaction was similar to that reported for tomatidine, the known spirosolane-type Solanum alkaloid. The structure elucidation of both metabolites was based primarily on 1D- and 2D-NMR analyses. PMID- 11021640 TI - Biosynthesis of tetrahydroanthraquinones in fungi. AB - The tetrahydroanthraquinones (1S,3S)-austrocortilutein (4) and (1S,3S) austrocortirubin (7) are octaketides formed in the fruit bodies of the fungus Dermocybe splendida via the gentiobioside 2 of the dihydroanthracenone (S) torosachrysone (1). PMID- 11021641 TI - Two polyisoprenylated benzophenones from the trunk latex of Clusia grandiflora (Clusiaceae). AB - The polyisoprenylated benzophenones, chamones I and II, were isolated from the trunk latex of Clusia grandiflora (Clusiaceae) growing in southeastern Venezuela. A third benzophenone, nemorosone II, was isolated from the pollinator reward resin of the female flowers of the same plant. Chamone I and nemorosone II are structurally similar, differing only in the degree of prenylation. Bioassays of chamone I and nemorosone II using the honeybee pathogens, Paenibacillus larvae and Paenibacillus alvei, demonstrate that both have potent antibacterial activity, and that their structural differences affect both their bactericidal efficacies and their aqueous mobilities. PMID- 11021642 TI - New bioactive aromatic compounds from Vismia guianensis. AB - Five benzophenones, vismiaguianones A-E, and two benzocoumarins, vismiaguianins A and B were isolated from the CHCl3 extract of the roots of Vismia guianensis by bioassay-directed fractionation using the DNA strand-scission assay and KB cell line. Of the isolates obtained, vismiaguianone B exhibited DNA strand-scission activity, whereas vismiaguianones D and E and vismiaguianin A were found to be significantly cytotoxic. PMID- 11021643 TI - (S)- and (R)-eriodictyol-6-C-beta-D-glucopyranoside, novel keys to the fermentation of rooibos (Aspalathus linearis). AB - The processed leaves and stems of Aspalathus linearis contain a new diastereomeric pair of the flavanones, (S)- and (R)-eriodictyol-6-C-beta-D glucopyranoside, which is also formed via the oxidative cyclization of the dihydrochalcone, aspalathin, under conditions which mimic the fermentation process. PMID- 11021644 TI - Antifungal monoterpene production in elicited cell suspension cultures of Piqueria trinervia. AB - Cell suspension cultures of the traditional medicinal plant Piqueria trinervia Cav., which synthesizes monoterpene piquerol A, were established. A defense response was induced in the cultures when eight homogenized fungi isolated from wild populations of P. trinervia were added. Piquerol A was not produced in the elicited system, while four other substances were synthesized de novo. They were excreted into the medium and inhibited in vitro fungal growth. The most abundant substance produced in this system was a new monoterpene: 2-methylene-7,7 dimethylbicyclo (3,3,1) heptane-4,6-diol. Monoterpenes in the cell suspension culture reported here were produced via two metabolic channels: the first acted constitutively and expressed in liquid and solid cultures, the second is inducible in response to several pathogens and elicitor substances. PMID- 11021645 TI - Triacontanol inhibits both enzymatic and nonenzymatic lipid peroxidation. AB - The effect of the plant growth regulator, triacontanol (TRIA) on lipid peroxidation was studied in three different systems: (i) isolated chloroplasts of spinach (Spinacea oleracea L.) leaves; (ii) egg lecithin liposomes; and (iii) soybean lipoxygenase (LOX) system. The nonenzymatic lipid peroxidation in isolated chloroplasts and egg lecithin liposomes was measured as the amount of thiobarbituric acid reactive substances (TBARS) formed. Inhibition of Fe2+ and/or light-induced lipid peroxidation by TRIA was observed in both isolated chloroplasts and egg lecithin liposomes. The kinetics of soybean lipoxygenase-1 (LOX-1) was studied using linoleic acid as the substrate. The enzyme was competitively inhibited by TRIA. The Ki for TRIA inhibition of the enzyme was estimated to be 3.2-5.0 microM according to different methods of estimation. TRIA has been known to exhibit anti-inflammatory action in animals and this anti inflammatory effect of TRIA might be mediated through inhibition of lipid peroxidation. Since LOX inhibitors have been extensively used as therapeutic agents, TRIA, being a natural compound has been suggested to be an effective anti inflammatory drug. PMID- 11021646 TI - Phenolic glycosides of forage legume Onobrychis viciifolia. AB - A chemical examination of the extractives of the leaves of sainfoin was undertaken as part of a programme directed at understanding the factors which may contribute to its nutritive value as animal feed. Among the low molecular weight phenolic compounds characterized were seven cinnamic acid derivatives and nine flavonoid glycosides all of which were identified by NMR spectroscopy. Included among these compounds were two new natural hydroxycinnamic esters namely methyl 6 O-p-trans-coumaroyl-beta-D-glucopyranoside and methyl 6-O-p-cis-coumaroyl-beta-D glucopyranoside and a novel flavonoid chrysoeriol-4'-O-(6''-O-acetyl)-beta-D glucopyranoside. PMID- 11021647 TI - Changes in grape seed polyphenols during fruit ripening. AB - The quantity and characterization of extracted flavan-3-ol monomers and procyanidins was determined in seeds from Vitis vinifera cv. Cabernet Sauvignon berries, over the course of ripening and at different levels of vine water status. The per berry extractive yield of all polyphenols decreased with maturity, and followed second-order kinetics. The flavan-3-ol monomers decreased most rapidly, followed by the procyanidin extension units and finally, the terminal units. The relative proportion of procyanidin extension units did not vary with maturity. During fruit ripening, the mean degree of polymerization of extracted procyanidins is unchanged when analyzed intact by HPLC, but decreases by thiolytic degradation. The proportion of extracted procyanidins resistant to acid catalyzed thiolysis increased with maturity. Changes in vine water status affected polyphenol amounts, indicating that cultural practices can be used to influence composition. Oxidation of the seed polyphenols during fruit ripening, could explain these observations. PMID- 11021648 TI - Atomic force microscopy analysis of intermediates in cobalt hexammine-induced DNA condensation. AB - The packaging pathway of cobalt hexammine-induced DNA condensation on the surface of mica was examined by varying the concentration of Co(NH3)6(3+) in a dilute DNA solution and visualizing the condensates by atomic force microscopy (AFM). Images reveal that cobalt hexammine-induced DNA condensation on mica involves well defined structures. At 30 microM Co(NH3)6(3+), prolate ellipsoid condensates composed of relatively shorter rods with linkages between them are formed. At 80 microM Co(NH3)6(3+), the condensed features include toroids with average diameter of approximately 240 nm as well as U-shaped and rod-like condensates with nodular appearances. The results imply that the condensates, whether toroids, U-shaped or rod-like structures have similar intermediate state which includes relatively shorter rod-like segments. The average size of the condensed toroids after incubated at room temperature for 5 h (approximately 240 nm) is much larger than that incubated for 0.5 h (approximately 100 nm). The results indicate that the condensation of DNA by Co(NH3)6(3+) is a kinetic-controlled process. PMID- 11021649 TI - Effect of coordinated ions on structure and flexiblity of parallel G quandruplexes: a molecular dynamics study. AB - Single tract guanine residues can associate to form stable parallel quadruplex structures in the presence of certain cations. Nanosecond scale molecular dynamics simulations have been performed on fully solvated fibre model of parallel d(G7) quadruplex structures with Na+ or K+ ions coordinated in the cavity formed by the 06 atoms of the guanine bases. The AMBER 4.1 force field and Particle Mesh Ewald technique for electrostatic interactions have been used in all simulations. These quadruplex structures are stable during the simulation, with the middle four base tetrads showing root mean square deviation values between 0.5 to 0.8 A from the initial structure as well the high resolution crystal structure. Even in the absence of any coordinated ion in the initial structure, the G-quadruplex structure remains intact throughout the simulation. During the 1.1 ns MD simulation, one Na+ counter ion from the solvent as well as several water molecules enter the central cavity to occupy the empty coordination sites within the parallel quadruplex and help stabilize the structure. Hydrogen bonding pattern depends on the nature of the coordinated ion, with the G-tetrad undergoing local structural variation to accommodate cations of different sizes. In the absence of any coordinated ion, due to strong mutual repulsion, 06 atoms within G-tetrad are forced farther apart from each other, which leads to a considerably different hydrogen bonding scheme within the G-tetrads and very favourable interaction energy between the guanine bases constituting a G-tetrad. However, a coordinated ion between G-tetrads provides extra stacking energy for the G-tetrads and makes the quadruplex structure more rigid. Na+ ions, within the quadruplex cavity, are more mobile than coordinated K+ ions. A number of hydrogen bonded water molecules are observed within the grooves of all quadruplex structures. PMID- 11021650 TI - Effect of neighboring bases on base-pair stacking orientation: a molecular dynamics study. AB - It is generally believed that base-pair stacking interaction in DNA double helix is one of the strongest interactions that governs sequence directed structural variability. However, X-ray crystal structures of some base-paired doublet sequences have been seen to adopt different structures when flanked by different base-pairs. DNA crystal database, however, is still too small to make good statistical inference about effect of such flanking residues. Influence of neighboring residue on the local helical geometry of a base-paired doublet in B DNA has been investigated here using molecular dynamics simulation. We have generated ensembles of structures for d(CA).d(TG) and d(AA).d(TT) base-paired doublets located at the centers of d(CGCGCAAAGCG).d(CGCTTTGCGCG) and d(CGCGAAAACGCG).d(CGCGTTTTCGCG) sequences along with their analogs by varying the bases either at 5'- or 3'- position to the central doublet. Comparison of base paired doublet parameters for the ensembles of structures show that stacking geometry of d(CA).d(TG) doublet depends on some of the flanking base-pairs. On the other hand d(AA).d(TT) doublet remains nearly unperturbed when the flanking residues are altered. PMID- 11021651 TI - The solution structure of a DNA duplex containing a single 1-(2-O,3-C-ethylene beta-D-arabinofuranosyl)thymidine nucleoside. AB - The structure of a DNA duplex containing one 1-(2-O,3-C-ethylene-beta-D arabinofuranosyl)-thymidine nucleoside (T5) modification was investigated by use of two-dimensional 1H NMR spectroscopy at 750 MHz. The structure of the d(CCGCT5AGCG):d(CGCTAGCGG) duplex (CT5AG) containing one of this 2'-O,3'-C-linked bicycloarabino conformational restricted modification has been determined. We obtained inter-proton distance bounds from NOESY spectra by including a complete relaxation matrix analysis. These distance bounds were used as restraints in molecular dynamics (rMD) calculations. We also analyzed the fine structure of the cross peaks in a selective DQF-COSY spectra to determine the sugar conformations of the nucleotides. Forty final structures were generated for CT5AG from A-form and B-form dsDNA starting structures. The root-mean-square deviation (RMSD) of the coordinates for the forty structures of the complex was 0.92A. The structures were observed to be markedly irregular compared to canonical B-DNA, especially in terms of large variations in propeller twist and buckle. Also, lack of stacking of two bases near the modification site is observed. The sugar conformations of all the unmodified nucleotides are close to pure C2'-endo conformation. The structural feature of CT5AG was discussed in relation to the thermal stability and resistance towards exonucleolytic degradation. PMID- 11021653 TI - Invariant and variable base stacking geometries in B-DNA and A-DNA. AB - We calculated the interatomic distances between all couples of non-hydrogen atoms belonging to the neighboring Watson-Crick base pairs in the available crystal structures of DNA. Their standard deviations revealed remarkably large differences in the variability of the base stacking geometries of the particular steps. In line with experimental studies in solution, (CpA)-(TpG) and (TpA).(TpA) were identified as the most variable or flexible steps in the crystal structures of B-DNA. On the other hand, base stacking geometries of the (ApT).(ApT) steps were the most invariant, which was very surprising because all three steps composed only of C and G were much more flexible. This finding suggests that conformational stability of DNA and the rigidity have different origins. Furthermore, the nucleotide sequence dependence of the flexibility was almost reversed in A-DNA because the most flexible steps in B-DNA were the least flexible in A-DNA. The most invariant steps of B-DNA were variable in A-DNA. The (ApT).(ApT) step was a notable exception to this rule because it belonged to the most rigid steps in both B-DNA and A-DNA. The present results are fully consistent with the properties that poly(dA-dT).poly(dA-dT), poly(dA).poly(dT), poly(dAdC).poly(dG-dT) and poly(dA-dG).poly(dC-dT) exhibit in solution. PMID- 11021652 TI - Binding of symmetrical cyanine dyes into the DNA minor groove. AB - Optical methods, such as fluorescence, circular dichroism and linear flow dichroism, were used to study the binding to DNA of four symmetrical cyanine dyes, each consisting of two identical quinoline, benzthiazole, indole, or benzoxazole fragments connected by a trimethine bridge. The ligands were shown to form a monomer type complex into the DNA minor groove. The complex of quinoline containing ligand with calf thymus DNA appeared to be the most resistant to ionic strength, and it did not dissociate completely even in 1 M NaCl. Binding of cyanine dyes to DNA could also be characterized by possibility to form ligand dimers into the DNA minor groove, by slight preference of binding to AT pairs, as well as by possible intercalation between base pairs of poly(dG)-poly(dC). The correlation found between the binding constants to DNA and the extent of cyanine dyes hydrophobicity estimated as the n-octanol/water partition coefficient is indicative of a significant role of hydrophobic interactions for the ligand binding into the DNA minor groove. PMID- 11021654 TI - Sequencing by hybridization with the generic 6-mer oligonucleotide microarray: an advanced scheme for data processing. AB - DNA sequencing by hybridization was carried out with a microarray of all 4(6) = 4,096 hexadeoxyribonucleotides (the generic microchip). The oligonucleotides immobilized in 100 x 100 x 20-microm polyacrylamide gel pads of the generic microchip were hybridized with fluorescently labeled ssDNA, providing perfect and mismatched duplexes. Melting curves were measured in parallel for all microchip duplexes with a fluorescence microscope equipped with CCD camera. This allowed us to discriminate the perfect duplexes formed by the oligonucleotides, which are complementary to the target DNA. The DNA sequence was reconstructed by overlapping the complementary oligonucleotide probes. We developed a data processing scheme to heighten the discrimination of perfect duplexes from mismatched ones. The procedure was united with a reconstruction of the DNA sequence. The scheme includes the proper definition of a discriminant signal, preprocessing, and the variational principle for the sequence indicator function. The effectiveness of the procedure was confirmed by sequencing, proofreading, and nucleotide polymorphism (mutation) analysis of 13 DNA fragments from 31 to 70 nucleotides long. PMID- 11021655 TI - Basic properties of populations generated in the frame of one-parameter discrete model of genetic diversity. AB - Previously, when discussing the properties of one parameter discrete model of genetic diversity (M.Yu. Shchelkanov et al, J. Biomol. Struct. Dyn. 15, 887-894 (1998)), we took into account Hamming distance distribution only between precursor and arbitrary descendant sequences. However, really there are sets of sequence populations produced during amplification process. In the presented work we have investigated Hamming distance distributions between sequences from different descendant sets produced in the frame of one parameter discrete model. Two basic descendant generation operators (so called amplifiers) are introduced: 1) the last generation amplifier, L, which produces descendants with precursor elimination; 2) all generations amplifier, G, which produces descendants without precursor elimination. Generalization of one-parameter discrete model for the case when precursor sequences do not coincide are carried out. Using this generalization we investigate the distribution of Hamming distances between L- and G-generated sequences. Basic properties of L and G operators, L/G-choice alternative problem have been discussed. Obtained results have common theoretical significance, but they are more suitable for high level genetic diversity process (for example, HIV diversity). PMID- 11021656 TI - The identification of tryptophan residues responsible for ATP-induced increase in intrinsic fluorescence of myosin subfragment 1. AB - ATP binding to myosin subfragment 1 (S1) induces an increase in tryptophan fluorescence. Chymotryptic rabbit skeletal S1 has 5 tryptophan residues (Trp113, 131, 440, 510 and 595), and therefore the identification of tryptophan residues perturbed by ATP is quite complex. To solve this problem we resolved the complex fluorescence spectra into log-normal and decay-associated components, and carried out the structural analysis of the microenvironment of each tryptophan in S1. The decomposition of fluorescence spectra of S1 and S1-ATP complex revealed 3 components with maxima at ca. 318, 331 and 339-342 nm. The comparison of structural parameters of microenvironment of 5 tryptophan residues with the same parameters of single-tryptophan-containing proteins with well identified fluorescence properties applying statistical method of cluster analysis, enabled us to assign Trp595 to 318 nm, Trp440 to 331 nm, and Trp 13, 131 and 510 to 342 nm spectral components. ATP induced an almost equal increase in the intensities of the intermediate (331 nm) and long-wavelength (342 nm) components, and a small decrease in the short component (318 nm). The increase in the intermediate component fluorescence most likely results from an immobilization of some quenching groups (Met437, Met441 and/or Arg444) in the environment of Trp440. The increase in the intensity and a blue shift of the long component might be associated with conformational changes in the vicinity of Trp510. However, these conclusions can not be extended directly to the other types of myosins due to the diversity in the tryptophan content and their microenvironments. PMID- 11021657 TI - Structure of glutamate decarboxylase and related PLP-enzymes: computer-graphical studies. AB - Amino acid sequences of E. coli glutamate decarboxylase (GADa) and those of 36 GAD of different origin were compared by pairwise alignment using computer program CLUSTAL. GADalpha and plant enzymes showed 59.8-67.8% subunit homology, GADalpha and other bacterial GAD--49.8-77.6%, whereas GADalpha and animal enzymes -13.9-58.8%. Two PLP domains exhibited higher homology comparing to that of the whole subunit in the case of GAD67, plant (68.4-73.9%), and bacterial (46.7 83.2%) enzymes. The alignment of PLP-domains of 37 GAD, three group II decarboxylases, and two pyridoxal enzymes with known 3D structures (bacterial ORD and mAAT from chicken heart) allowed us to reveal conserved residues of the active sites. Their functional role is discussed. Modelling of the PLP-binding sites in active centers for GADalpha and human brain GAD67 was done using the Swiss-PdbViewer homology modelling program. Although the homology between GADalpha and GAD67 is rather low, structural similarity of their active sites allows us to consider here a functional convergence. Thus, glutamate decarboxylation by GADalpha may be helpful for understanding general mechanism of this reaction. PMID- 11021658 TI - Homology model of a novel xylanase: molecular basis for high-thermostability and alkaline stability. AB - Xylanases form enzymes of considerable interest to a variety of biotechnological industries. Their industrial usage is especially attractive since they can replace some of the environmental pollutants, and are economically viable. Those with higher thermostability and optimal activity at alkaline pH are of particular importance to the paper and pulp industry due to the demands of conditions under which the enzymatic reactions are carried out. We have earlier isolated a xylanase from Bacillus sp. NG-27, which is active both at high temperature as well as at alkaline pH. In order to find out factors responsible for the adaptation of this enzyme to the extreme conditions, three dimensional structure of NG-27 xylanase has now been obtained by homology modelling. The tertiary structure shows TIM barrel fold consisting of 8 parallel beta-strands surrounded by alpha-helices. The active site is located at the carboxy terminal end of the TIM barrel. Factors which contribute to the thermostability of the enzyme are increased number of salt bridges. The salt bridges occur remarkably on one face of alpha-helices, with oppositely charged residues occupying i, i+4, i+7 positions. A solvent shielded salt bridge interaction is also observed, which is absent in the mesophilic homologous xylanases. Solvent shielding may enhance electrostatic interaction through lowering of the dielectric, and contribute to increased stability of the enzyme. PMID- 11021659 TI - Solution structure of O-glycosylated C-terminal leucine zipper domain of human salivary mucin (MUC7). AB - Solution structures of a 23 residue glycopeptide II (KIS* RFLLYMKNLLNRIIDDMVEQ, where * denotes the glycan Gal-beta-(1-3)-alpha-GalNAc) and its deglycosylated counterpart I derived from the C-terminal leucine zipper domain of low molecular weight human salivary mucin (MUC7) were studied using CD, NMR spectroscopy and molecular modeling. The peptide I was synthesized using the Fmoc chemistry following the conventional procedure and the glycopeptide II was synthesized incorporating the O-glycosylated building block (Nalpha-Fmoc-Ser-[Ac4-beta-D-Gal (1,3)-Ac2-alpha-D-GalN3+ ++]-OPfp) at the appropriate position in stepwise assembly of peptide chain. Solution structures of these glycosylated and nonglycosylated peptides were studied in water and in the presence of 50% of an organic cosolvent, trifluoroethanol (TFE) using circular dichroism (CD), and in 50% TFE using two-dimensional proton nuclear magnetic resonance (2D 1H NMR) spectroscopy. CD spectra in aqueous medium indicate that the apopeptide I adapts, mostly, a beta-sheet conformation whereas the glycopeptide II assumes helical structure. This transition in the secondary structure, upon glycosylation, demonstrates that the carbohydrate moiety exerts significant effect on the peptide backbone conformation. However, in 50% TFE both the peptides show pronounced helical structure. Sequential and medium range NOEs, CalphaH chemical shift perturbations, 3JNH:CalphaH couplings and deuterium exchange rates of the amide proton resonances in water containing 50% TFE indicate that the peptide I adapts alpha-helical structure from Ile2-Val21 and the glycopeptide II adapts alpha-helical structure from Ser3-Glu22. The observation of continuous stretch of helix in both the peptides as observed by both NMR and CD spectroscopy strongly suggests that the C-terminal domain of MUC7 with heptad repeats of leucines or methionine residues may be stabilized by dimeric leucine zipper motif. The results reported herein may be invaluable in understanding the aggregation (or dimerization) of MUC7 glycoprotein which would eventually have implications in determining its structure-function relationship. PMID- 11021660 TI - Density functional geometry optimization and energy calculations of calcium(II) triphosphate complexes. Polyphosphates as possible dissolving agents for calcium pyrophosphate dihydrate crystals in chondrocalcinosis disease. AB - Geometry optimizations and energy calculations have been carried out via molecular orbital methods at the density functional B3LYP/LANL2DZ level on the molecules PO3-, OPO3(3-), HOPO3(2-), CH3OPO3(2-), H(CH3OPO3)-, O(PO3)2(4-), HO(PO3)2(3-), CH2(PO3)2(4-), (CH3OPO2)O(PO3)3-, O(PO3)3(5-), HO(PO3)3(4-), (PO3)3(3-), (CH3OPO2)O(PO3)2(4-), [Mg[O(PO3)2)]]2-, [Ca[O(PO3)2]]2-, [Ca[CH2(PO3)2]]2-, [Ca[CH3OPO2)O(PO3)]]-, [Ca(PO3)3]-, [Ca[O(PO3)3]]3-, and [Ca[CH3OPO2)O(PO3)2]]2- with the aim to find reliable and easily accessible computational methods to simulate some phosphate-containing molecules of importance for the living cells and to study the energetics for protonation and metal-complex formation reactions. The analysis is part of a general investigation on phosphate-containing molecules as potential dissolving agents for calcium pyrophosphate dihydrate (CPPD) crystals which deposit in certain articular diseases. The basis set was expanded to 6-31G** for the P atoms for all the molecules investigated and to 6-31G* for the O atoms for OPO3(3-). Calculations at the semiempirical MNDO/d level were also carried out for comparison purposes on the free ligand molecules and on [Mg[O(PO3)2]]2-. The density functional analysis reproduced well the geometry found at the solid state via X-ray diffraction. The analyses of the geometrical parameters and the total electronic energy of the molecules shows that O(PO3)2(4-) and other di- and tri phosphates are versatile ligands for divalent metal ions like Ca2+. The computed P-O-P bond angle for free O(PO3)2(4-) is 180 degrees and the conformation of the two PO3- groupings is staggered along the P...P vector. The linear arrangement for P-O-P is assisted by P-O pi interactions. The bending of the P-O-P angle when accompanied by a slight P-O(b) elongation requires a very small amount of energy; 4.65 kcal/mol to pass from 180 to 140 degrees , as calculated at the DFT level. The computed Ca-O and Mg-O bond distances for [M[O(PO3)2]]2- are 2.378 and 2.079A, when the metal ions link two oxygen atoms from each PO3 group. The computed Ca-O bond lengths for [Ca[CH3OPO2)O(PO3)]]- are 2.482 (PalphaO2) and 2.358A (PbetaO2), showing a significant lengthening for Ca-OPalpha, when compared to the pyrophosphate derivative. The Ca-O bond lengths for [Ca[O(PO3)3]]3- and [Ca[CH3OPO2)O(PO3)2]]2- are 2.251A and 2.525 (PalphaO2), 2.407 and 2.338 (PbetaO2), and 2.251 and 2.228A (PgammaO2), showing a shortening for the Ca OPgamma bond upon methylation. The (Pbeta)O-Pgamma bond length increases significantly (0.09 A) upon Ca(II) coordination to (CH3OPO2)O(PO3)2(4-) via all the three PO3 groups. This latter result suggests that metal complexes of linear organic-triphosphates have a larger tendency to release the PgammaO3 group when compared to the free ligand molecules. The electronic contribution to the energy of the complex formation reaction for [Ca[CH2(PO3)2]]2- is only slightly higher (some 1.8 kcal) than that for [Ca[O(PO3)2]]2-; but is much higher (some 63 kcal) than that relevant to the formation of [Ca[CH3OPO2)O(PO3)2]]2-. (ABSTRACT TRUNCATED) PMID- 11021661 TI - Disordered regulation of coagulation and platelet activation in xenotransplantation. AB - Rejection of xenografts is associated with vascular-based inflammation, thrombocytopenia and the consumption of coagulation factors that may evolve into disseminated intravascular coagulation (DIC). Similarly, bone marrow-derived cellular xenotransplantation procedures are associated with endothelial cell activation and thrombotic microangiopathic injury. These complications generally develop despite the best available measures for depletion of xenoreactive natural antibody, inhibition of complement activation and suppression of T- and B-cell mediated immune responses. The mechanisms underlying the DIC and thrombotic microangiopathy associated with xenotransplantation are unclear. A proposed primary biological dysfunction of xenografts with respect to regulation of clotting could amplify vascular injury, promote immunological responses and independently contribute to graft failure. Disordered thromboregulation could have deleterious effects, comparable to unregulated complement activation, in the pathogenesis of xenograft rejection and may therefore represent a substantive barrier to xenotransplantation. PMID- 11021662 TI - Lack of antibody production against Hanganutziu-Deicher (H-D) antigens with N glycolylneuraminic acid in patients with porcine exposure history. AB - The significance of non-alphagalactosyl antigens remains unclear in pig-to primate xenotransplantation. Hanganutziu-Deicher (H-D) antigens with terminal N glycolylneuraminic acid (NeuGc) are widely expressed on endothelial cells of mammalian species, with the exception of humans. As baboons and monkeys also express H-D antigens, a pig-to-non-human primate experimental model cannot resolve the question of whether H-D antigens can elicit a potent humoral response in human recipients. The purpose of this study was to elucidate the clinical significance of H-D antigens by examining the sera from patients who have been previously exposed to porcine tissue. After the digestion of porcine aortic endothelial cells (PAEC) by neuraminidase, NeuGc and N-acetylneuraminic acid (NeuAc) were quantitated by HPLC. IgG and IgM antibody levels against H-D antigens were measured by NeuGc-GM3-coated ELISA plates in the sera of patients who had undergone ex vivo kidney perfusion 1 to 3 weeks and 2 years previously (n=2) or had been injected with fetal porcine islets 2 months previously (n= 10). HPLC determined that 9.7x 10(7) NeuAc and 6.3x 10(7) NeuGc residues per cell were released from PAEC by neuraminidase, while 25.7x 10(7) NeuAc and an undetectable level of NeuGc were released from human aortic endothelial cells (HAEC). No significant elevation of IgG or IgM antibody levels against NeuGc-GM3 was observed in sera from patients with a history of porcine exposure. Considering the active production of antibody against the foreign galactosyl antigens after pig-to-human xenotransplantation, some production of antibodies against the equally foreign H-D antigens would be expected, because large amounts of NeuGc terminated saccharides are present in the pig endothelial cell surface. However, no production of antibodies directed to H-D antigens could be found in patients exposed to porcine tissue. Further studies are warranted to explain why H-D antigens do not elicit a significant antibody production. PMID- 11021663 TI - Plasma perfusion by apheresis through a Gal immunoaffinity column successfully depletes anti-Gal antibody: experience with 320 aphereses in baboons. AB - BACKGROUND: Anti-Galalpha1-3Gal (Gal) antibodies (Gal Ab) contribute to the rejection of porcine organs transplanted into primates. Extracorporeal immunoadsorption (EIA) has been developed to eliminate Gal Ab from the circulation. METHODS: Between 1995 and 1999 we performed 320 EIAs in baboons using a COBE-Spectra apheresis unit incorporating a synthetic Gal immunoaffinity column. Three plasma volumes were immunoadsorbed on each occasion. The 221 consecutive EIAs performed in 41 immunosuppressed baboons between January 1997 and April 1999 form the basis of this review. Of these 41 baboons, 29 underwent a series of three or four EIAs at daily intervals, seven had multiple series of three EIAs, and the remainder underwent single or double EIAs. Serum Gal Ab levels were monitored by ELISA before and at intervals after the course of EIA. RESULTS: There were two fatal complications, one from a respiratory mishap (unrelated to the EIA) and one from persistent hypotension unresponsive to therapeutic interventions. Seven procedures (3%) were terminated early owing to technical difficulties and/or persistent hypotension. Mean pre-EIA Gal Ab levels in naive baboons were 33.1 microg/ml (IgM) and 14.5 microg/ml (IgG). Immediately after three consecutive EIAs, IgM was depleted by a mean of 97.3% and IgG by 99.4%. By 18 to 24 h later, Gal Ab was returning but depletion remained at 80.1% (IgM) and 84.7% (IgG). The subsequent rate of return of Gal Ab depended on the immunomodulatory protocol used. CONCLUSIONS: (1) With appropriate monitoring, EIA is an acceptably safe procedure, even in small (<10 kg) baboons. (2) Three consecutive EIAs are effective in removing >97% of Gal Ab. (3) In the majority of cases, return of Gal Ab begins within 24 h, irrespective of the immunomodulatory protocol. PMID- 11021664 TI - Acute vascular rejection is associated with systemic complement activation in a pig-to-primate kidney xenograft model. AB - The introduction of h-DAF transgenic porcine organs into pre-clinical pig-to primate discordant xenotransplantation has led to complete and reliable abrogation of hyperacute xenograft rejection (HAR). Despite additional heavy immunosuppression however, most xenografts are still lost due to acute vascular rejection (AVR), with current treatment protocols being of only limited value. In a life-supporting model of pig-to-primate kidney transplantation, unmodified (n=8) or h-DAF-transgenic (n=9) porcine kidneys were transplanted into cynomolgus monkeys under cyclophosphamide (CyP), cyclosporine and low-dose steroid immunosuppression. Longest recipient survival was 11 days in the control group and 68 days in the h-DAF transgenic group. Stable initial graft function with recipient survival >4 days was generated in eight animals (two controls and six transgenics). In these animals, plasma complement levels were analyzed during ongoing AVR. Compared with baseline levels, a two-fold increase in C3a levels and a four-fold increase in sC5b-9 levels were measured. In parallel to systemic complement activation, increased deposition of C3 and C5b-9 along with massive staining for recipient IgM immunoglobulins was detected in the xenografts on immunohistochemistry. We conclude that acute vascular xenograft rejection of porcine kidneys in cynomolgus monkeys is associated with classical pathway complement activation following binding of induced recipient anti-porcine antibodies. This complement activation can be observed despite membrane bound expression of human complement regulators in the porcine xenografts. Therefore, additional short-term fluid phase complement inhibition seems necessary for the future development of protocols designed for treatment of AVR in the pig-to primate combination. PMID- 11021665 TI - Inducible nitric oxide synthetase is expressed in adult but not fetal pig pancreatic islets. AB - Cytokine-induced expression of inducible nitric oxide synthetase (iNOS) and production of nitric oxide (NO) by pancreatic islet cells has been suggested as one potential mechanism for beta cell destruction. In this study, we investigated the role of iNOS and NO in islet primary non-function. Islets were assessed for their function, viability and expression of iNOS. Adult rat and pig islets isolated by collagenase digestion and fetal pig pancreas (FPP) grafts isolated by collagenase digestion or high oxygen culture were transplanted into C57BL6 mice and nude mice. iNOS protein was detected by immunohistochemistry. iNOS protein was found in normal rat and pig pancreas and adult rat and pig islets that were isolated by collagenase digestion and transplanted into either C57BL6 mice or nude mice. iNOS was not detected in fetal pig islet grafts, regardless of whether collagenase was used in the isolation process. In adult pig islet grafts, the presence of iNOS protein correlated with high levels of islet cell apoptosis and primary non-function. Despite the persistent presence of iNOS in rat islets, there was no evidence that it had a deleterious effect on rat islet viability, or function. Therefore, in isolated adult pig islets, there was a correlation between iNOS expression and apoptosis, suggesting that iNOS activation may be deleterious to the adult pig islets. However, other factors such as the fragility of the islet capsule may be equally important. By contrast, fetal pig islets did not express iNOS and this may be an important reason for their enhanced viability when compared with adult islet tissue. PMID- 11021666 TI - Delayed type hypersensitivity-associated cytokines in islet xenotransplantation: limited efficacy of interleukin-2- and tumor necrosis factor-alpha-blockade in interferon-gamma receptor-deficient mice. AB - To investigate the role of interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha and their potential to replace each other in the process of fetal porcine islet-like cell cluster (ICC) xenograft rejection, mice with a targeted disruption of the IFN-gamma receptor gene and wild-type controls were transplanted with fetal porcine ICCs under the kidney capsule and given post transplant treatment with the TNF-alpha-inhibiting agent MDL 201,449A. Some of the MDL 201,449A-treated IFN-gamma receptor-deficient mice received additional treatment with cyclosporinee (CsA). Evaluation of the xenografts was performed 7 days after transplantation (all groups), and in IFN-gamma receptor-deficient mice treated with MDL 201 449 A, also 10 and 13 days after transplantation. On day 7 after transplantation, a few CD3+ cells were seen accumulated peripherally in the ICC xenograft. Moderate to abundant numbers of F4/80+ and Mac-1+ cells surrounded a few remaining ICCs present within the xenograft. Histochemical visualization of cyanide-resistant endogenous peroxidase activity for detection of eosinophils demonstrated only small numbers of eosinophils present within the xenograft by day 7 after transplantation. An increased amount of eosinophilic granulocytes was not found until day 10 after transplantation, i.e. at a time when ICC xenograft rejection has already been completed. However, two out of six IFN-gamma receptor deficient mice given post-transplant treatment with CsA and MDL 201,449A exhibited intact ICC xenografts with ICCs arranged in chords and duct-like structures on day 7 after transplantation. Taken together, findings in this study indicate that, in the pig-to-mouse model, IFN-gamma, TNF-alpha, and interleukin-2 seem to be of importance to fetal porcine ICC xenograft rejection. Nevertheless, in a majority of animals, other cytokines eventually substitute for the lack of IFN-gamma, TNF-alpha and interleukin-2. PMID- 11021667 TI - Pig islet xenograft rejection is markedly delayed in macrophage-depleted mice: a study in streptozotocin diabetic animals. AB - The present study aimed to evaluate the effect of depletion of macrophages and/or natural killer (NK) cells on islet xenograft rejection in the pig-to-mouse model. Five microliters (4,000 to 5,000 IEQ, islet equivalents) of adult pig islets were transplanted under the renal capsule of C57BL/6 mice with streptozotocin-induced diabetes. Macrophages were depleted by injection of liposome-encapsulated dichloromethylene diphosphonate (Lip-Cl2MDP) intraperitoneally (i.p.) at a dose of 100 microl/ 10 g body weight (BW) 2 days before transplantation, and 50 microl/10 g BW weekly thereafter. NK cells were depleted by injection of the monoclonal antibody NK 1.1 (anti-NK 1.1 mAb) i.p. at a dose of 100 microg/mouse 1 day before transplantation, and then 25 microg per week thereafter. Islet graft survival was monitored by daily measurements of blood glucose. Graft survival was 8 +/- 1.2 days in untreated controls, 9 +/- 1.0 days with anti-NK 1.1 mAb alone, 22 +/- 4.9 days with Lip-Cl2MDP alone (P<0.01 vs. controls), and 26 +/- 3.8 days with Lip-Cl2MDP plus anti-NK 1.1 mAb (P<0.01 vs. controls). In the last group, two of six animals were killed with functioning grafts 30 days after transplantation. In untreated controls, rejected xenografts were heavily infiltrated by F4/80+ macrophages and CD3+T cells. In Lip-Cl2MDP-treated groups, the number of F4/80+ macrophages was markedly reduced. On the periphery of xenografts, a small number of CD3+T cells were observed. In conclusion, our results suggest that strategies targeting macrophages may facilitate islet xenograft survival. A role for NK cells cannot be excluded, but appears to be of minor importance. PMID- 11021668 TI - Generation and testing of a highly specific anti-serum directed against porcine endogenous retrovirus nucleocapsid. AB - Advances in xenotransplantation offer chances to alleviate the shortage of human donor organs. The discovery that pig endogenous retroviruses (PERV) can infect human cells in vitro has stimulated the discussion on infectious risk in xenotransplantation. A molecular and immunologic monitoring of xenograft recipients and of donor animals for putative infection with PERV and other microorganisms is inevitable. In this report, we describe the generation and testing of a highly specific anti-serum directed against the PERV nucleocapsid protein. The Gag amino acid (aa) sequence of PERV class B was used to define immunogenic domains by computer analysis. A peptide corresponding to the C terminal 19 aa of the 10 kDa (p10) nucleocapsid (NC) portion of the Gag polyprotein was used to immunize rabbits. The generated serum was tested using recombinant PERV Gag protein expressed in insect cells, purified PERV virus particles and human 293 cells transfected or infected with PERV, respectively. Test methods included Western blotting, indirect immunofluorescence, immunoperoxidase assay and ELISA. The PERV anti-serum provides a tool that is instrumental for detection of a potential agent of zoonosis. It can be used for screening of donor animals and xenograft recipients in the course of xenotransplantation procedures. PMID- 11021669 TI - Literature update 2000, part 1. PMID- 11021670 TI - Should health plan quality measures be adjusted for case mix? PMID- 11021671 TI - Impact of sociodemographic case mix on the HEDIS measures of health plan quality. AB - BACKGROUND: The widely used Health Plan Employer Data and Information Set (HEDIS) measures may be affected by differences among plans in sociodemographic characteristics of members. OBJECTIVE: The objective of this study was to estimate effects of geographically linked patient sociodemographic characteristics on differential performance within and among plans on HEDIS measures. RESEARCH DESIGN: Using logistic regression, we modeled associations between age, sex, and residential area characteristics of health plan members and results on HEDIS measures. We then calculated the impact of adjusting for these associations on plan-level measures. SUBJECTS: This study included 92,232 commercially insured members with individual-level HEDIS data and an additional 20,615 members whose geographic distribution was provided. MEASURES: This study used 7 measures of screening and preventive services. RESULTS: Performance was negatively associated with percent receiving public assistance in the local area (6 of 7 measures), percent black (5 measures), and percent Hispanic (2 measures) and positively associated with percent college educated (6 measures), percent urban (2 measures), and percent Asian (1 measure) after controlling for plan and product type. These effects were generally consistent across plans. When measures were adjusted for these characteristics, rates for most plans changed by less than 5 percentage points. The largest change in the difference between plans ranged from 1.5% for retinal exams for people with diabetes to 20.2% for immunization of adolescents. CONCLUSIONS: Performance on quality indicators for individual members is associated with sociodemographic context. Adjustment has little impact on the measured performance of most plans but a substantial impact on a few. Further study with more plans is required to determine the appropriateness and feasibility of adjustment. PMID- 11021672 TI - Gender-related differences in the organization and provision of services among general practitioners in Europe: a signal to health care planners. AB - BACKGROUND: The number of women entering general practice is rising in many countries. Thus, gender differences in work situation preferences and practice activities are important for future planning. OBJECTIVES: This article describes the differences between male and female general practitioners (GPs) in 32 European countries. It examines gender differences in curative and preventive services and relates these to features of the health care system and the practice. METHODS: The data were collected in 1993 and 1994 in the European Study of Task Profiles of General Practitioners. In 32 countries, 8,183 GPs answered standardized questionnaires written in their own languages on their self-reported involvement in curative and preventive services, as well as how their practice was organized and managed. Because the independent variables in this study were on both the national 1 and individual practice levels, the data were subjected to multilevel analysis. RESULTS: Regardless of the type of health care system, the female GPs were younger than the male GPs and more often worked part time in groups or partnerships and in cities, although not in deprived areas. They made fewer house calls and did less work outside office hours. Differences between men and women regarding workload diminished considerably after controlling for part time work. When other characteristics of the person and the practice were taken into account, female GPs proved to be less involved in several curative services, except as the first contact for gynecological problems, but more involved in health education. Some differences were found in only certain types of health care systems. CONCLUSIONS: The results may have important implications for working arrangements, training, education, and planning of resources for general practice in the future. PMID- 11021673 TI - Hoptel equalizes length of stay for homeless and domiciled inpatients. AB - BACKGROUND: There is growing evidence that homeless individuals have longer inpatient lengths of stay with significantly higher medical costs than domiciled individuals. We compared adjusted mean lengths of stay among domiciled patients and homeless patients discharged to a hospital hotel (hoptel). Because the hoptel allowed homeless patients to be discharged when medically indicated, we hypothesized no significant differences in lengths of stay between the 2 groups after adjustment for severity of illness. METHODS: Demographic, diagnosis, and length of stay data were obtained for all patients discharged from all inpatient wards at a large, urban Veterans Affairs Medical Center during the first 32 months of hoptel operation (1996 through 1998). Adjusted mean lengths of stay for domiciled patients and homeless patients discharged to the hoptel were compared. ANCOVA was used to adjust for demographic characteristics (age, income, race/ethnicity), diagnosis-related group (DRG), principal diagnosis, and substance abuse comorbidity. Three types of hospitalizations were analyzed on the basis of major DRG categories: general medical, psychiatric, and all hospitalizations. RESULTS: No significant differences were found in adjusted mean lengths of stay between domiciled patients and homeless patients discharged to the hoptel. Homeless hoptel patients stayed an average 0.7, 0.3, and 0.4 days longer than domiciled patients for all, general medical, and psychiatric hospitalizations (P = 0.504, 0.627, and 0.839), respectively. CONCLUSIONS: These data suggest that hoptels may help equalize inpatient lengths of stay among domiciled and homeless patients discharged to a hoptel. Public health care systems seeking innovative ways of reducing inpatient lengths of stay among homeless patients ought to consider establishing hoptels. PMID- 11021674 TI - Differences in health values among patients, family members, and providers for outcomes in schizophrenia. AB - OBJECTIVE: The objectives of this study were to determine whether there are important differences in how patients, family members, and health care providers (HCPs) value health outcomes in schizophrenia and to assess the degree to which such differences, if they exist, could adversely affect clinical and policy decision making. METHODS: Participants viewed videotaped depictions of simulated patients with mild and moderate symptoms of schizophrenia, with and without a common adverse drug effect (pseudoparkinsonism), and then provided standard gamble and visual analog scale ratings of desirability of these states. SUBJECTS: A convenience sample of unrelated patients (n = 148), family members of patients (n = 91), and HCPs (nurses, psychologists, doctors of pharmacy, and doctors of medicine; n = 99) was drawn from geographically and clinically diverse environments. RESULTS: Patients' and family members' utilities for health states averaged 0.1 to 0.15 units higher than those of HCPs (P <0.002 for differences between groups, ANOVA for multiple observations). The disutility of adverse drug effects was less for health professionals than patients and family members (P = 0.008). Health professionals tended to prefer states with mild symptoms with extrapyramidal side effects to states with moderate symptoms. Patients and family members found these states equally preferable (P <0.007 for differences between groups). CONCLUSIONS: There are systematic differences in values for health outcomes between patients and HCPs with regard to states with adverse effects of antipsychotic drugs. Family members of patients in general had values that were more similar to those of patients than were those of health professionals. The results emphasize the importance of participation by patients (or family member proxies) in clinical decision making and guideline development. PMID- 11021675 TI - Performance of the SF-36, SF-12, and RAND-36 summary scales in a multiple sclerosis population. AB - BACKGROUND: Multiple sclerosis (MS) patients accumulate both physical and mental health problems along with disease progression. Valid and sensitive outcome measures are important to measure disease effects and the effect of treatment. OBJECTIVE: The objective of this study was to test the performance of the physical and mental summary scales of SF-36, SF-12, and RAND-36. METHODS: The scales were evaluated by comparing the scores of a cohort of 194 MS patients with general population data and using the Expanded Disability Status Scale (EDSS) and the Incapacity Status Scale-mental as criterion variables for physical functioning and mental health. RESULTS: All 3 physical summary scales were markedly reduced and correlated highly with the EDSS. The SF-36 mental summary score was only slightly reduced among MS patients (0.2 SD) compared with the general population, despite significantly reduced scores on all 4 health scales being most related to mental health and despite a high prevalence of mental health problems. This results from the poor physical functioning (mean scale score, 2.3 SD below the general population) and the orthogonal factor rotation used to derive independent measures of physical and mental health. Similar results were found for the SF-12. The nonorthogonal RAND-36 physical and mental summary scores were both markedly reduced. This is more compatible with the disease progression in MS and the results of the other measures of physical and mental health used in the study. CONCLUSIONS: The SF-36 and SF-12 mental health summary scales appear to overestimate mental health in people with MS. PMID- 11021676 TI - Physicians' responses to Medicare fee schedule reductions. AB - BACKGROUND: Relatively little empirical research has addressed physicians' responses to fee changes under the Medicare Fee Schedule. OBJECTIVES: We analyzed Medicare claims data for ophthalmologists and orthopedic surgeons for the years 1991 through 1994 to evaluate the relative importance of profit-maximizing and target-income theories in determining physicians' supply responses to specific Medicare fee reductions. RESEARCH DESIGN: This study was designed to estimate the impact of fee reductions for cataract extractions and major joint repair/replacement procedures through pooled cross-section time series data. RESULTS: The supply function for cataract extractions has both strong own-price and cross-price effects, as well as a highly significant negative income effect. Yet, the magnitude of the income effect is small; thus, the substitution effect dominates the income effect. Similarly, in the supply functions for joint procedures, the own price has the expected positive sign, implying that as the fee declines, orthopedic surgeons will perform fewer joint surgeries. However, the cross-price variable has the correct sign only if treated as exogenous, and the variables measuring the income effect have the wrong sign, although their magnitude is small. CONCLUSIONS: These results suggest that the Medicare Fee Schedule does have the potential to influence physicians' supply decisions, but these effects may vary by specialty and service. PMID- 11021677 TI - A trial for comparing methods for eliciting treatment preferences from men with advanced prostate cancer: results from the initial visit. AB - OBJECTIVE: The objective of this study was to evaluate the convergent validity of 3 types of utility measures: standard gamble, time tradeoff, and rating scale. RESEARCH DESIGN: A prospective cohort of 120 men with advanced prostate cancer were first asked to rank order 8 health states, and then utility values were obtained from each participant for each of the 8 health states through 2 of the 3 techniques evaluated (standard gamble, time tradeoff and rating scale). Participants were randomly assigned to 1 of 3 possible pairs of techniques. The validity of the 3 methods, as measured by the convergence and raw score differences of the techniques, was assessed with ANOVA. The ability of the techniques to differentiate health states was determined. The inconsistencies between rankings and utility values were also measured. Proportions of illogical utility responses were assessed as the percent of times when states with more symptoms were given higher or equal utility values than states with fewer symptoms. RESULTS: There were significant differences in raw scores between techniques, but the values were correlated across health states. Utility values were often inconsistent with the rank order of health states. In addition, utility assessment did not differentiate the health states as well as the rank order. Furthermore, utility values were often illogical in that states with more symptoms received equal or higher utility values than states with fewer symptoms. CONCLUSIONS: Use of the utility techniques in cost-effectiveness analysis and decision making has been widely recommended. The results of this study raise serious questions as to the validity and usefulness of the measures. PMID- 11021678 TI - Mobility problems and perceptions of disability by self-respondents and proxy respondents. AB - BACKGROUND: The Americans With Disabilities Act defines disability on the basis of physical or mental impairments or external perceptions of impairment. OBJECTIVES: The objective of this study was to examine perceptions of disability among people with lower-extremity mobility difficulties. RESEARCH DESIGN: This study used a cross-sectional, nationally representative survey, the 1994 to 1995 National Health Interview Survey-Disability (NHIS-D) supplement. Using SAS callable SUDAAN for all analyses, we produced national population estimates. SUBJECTS: This study included 142,572 noninstitutionalized, civilian residents of the United States who were > or =18 years of age, with 80,423 self-respondents and 49,883 proxy respondents. MEASURES: We created a 4-level mobility variable using NHIS-D questions about the ability to walk, climb stairs, stand and the use of mobility aids. We examined associations between mobility and answers to 2 questions about self- and external perceptions of disability. RESULTS: The results showed that 3.1% (estimated 5.82 million persons) reported major mobility difficulties, including 3.7% of self-respondents and 2.7% of those with proxy respondents. Among persons with major mobility problems, 70.8% perceived themselves as disabled, whereas 64.8% thought other people see them as disabled. Also, 80.5% of manual wheelchair users saw themselves as disabled. Proxies were somewhat more likely to perceive disability than self-respondents, although differences were not generally statistically significant. In multivariable regressions, mobility level was the strongest predictor of self-perceived disability, followed by general health status. CONCLUSIONS: Mobility problems increase the likelihood that people will see themselves as disabled, but these perceptions are not universal. Although the schematic of wheelchair users has become an international symbol of disability, many people with serious mobility problems do not view themselves as disabled. PMID- 11021679 TI - Volumetric reconstruction of medical images. PMID- 11021680 TI - Exact radon rebinning algorithm for the long object problem in helical cone-beam CT. AB - This paper addresses the long object problem in helical cone-beam computed tomography. We present the PHI-method, a new algorithm for the exact reconstruction of a region-of-interest (ROI) of a long object from axially truncated data extending only slightly beyond the ROI. The PHI-method is an extension of the Radon-method, published by Kudo, Noo, and Defrise in issue 43 of journal Physics in Medicine and Biology. The key novelty of the PHI-method is the introduction of a virtual object fpsi(x) for each value of the azimuthal angle psi in the image space, with each virtual object having the property of being equal to the true object f(x) in some ROI omegam. We show that, for each psi, one can calculate exact Radon data corresponding to the two-dimensional (2-D) parallel-beam projection of fpsi(x) onto the meridian plane of angle psi. Given an angular range of length pi of such parallel-beam projections, the ROI omegam can be exactly reconstructed because f(x) is identical to fpsi(x) in Omegam. Simulation results are given for both the Radon-method and the PHI-method indicating that 1) for the case of short objects, the Radon- and PHI-methods produce comparable image quality, 2) for the case of long objects, the PHI-method delivers the same image quality as in the short object case, while the Radon method fails, and 3) the image quality produced by the PHI-method is similar for a large range of pitch values. PMID- 11021681 TI - Exact (spiral + circles) scan region-of-interest cone beam reconstruction via backprojection. AB - We present a (spiral + circles) scan cone beam reconstruction algorithm in which image reconstruction proceeds via backprojection in the object space. In principle, the algorithm can reconstruct sectional region-of-interest (ROI) in a long object. The approach is a generalization of the cone beam backprojection technique developed by Kudo and Saito in two aspects: the resource-demanding normalization step in the Kudo and Saito's algorithm is eliminated through the technique of data combination that we published earlier, and the elimination of the restriction that the detector be big enough to capture the entire cone beam projection of the ROI. Restricting the projection data to the appropriate angular range required by data combination can be accomplished by a masking process. Because of the simplification resulting from the elimination of the normalization step, the most time-consuming operations of the algorithm can be approximated by the efficient step of line-by-line ramp filtering the cone beam image in the direction of the scan path, plus a correction image. The correction image, which can be computed exactly, is needed because data combination is not properly matched at the mask boundary when ramp filtering is involved. Empirical two dimensional (2-D) point spread function (PSF) is developed to improve matching with the correction image which is computed with finite samplings. The use of transition region to further improve matching is introduced. The results of testing the algorithm on simulated phantoms are presented. PMID- 11021682 TI - Multislice helical CT: image temporal resolution. AB - A multislice helical computed tomography (CT) halfscan (HS) reconstruction algorithm is proposed for cardiac applications. The imaging performances (in terms of the temporal resolution, z-axis resolution, image noise, and image artifacts) of the HS algorithm are compared to the existing algorithms using theoretical models and clinical data. A theoretical model of the temporal resolution performance (in terms of the temporal sensitivity profile) is established for helical CT, in general, i.e., for any number of detector rows and any reconstruction algorithm used. It is concluded that the HS reconstruction results in improved image temporal resolution than the corresponding 180 degrees LI (linear interpolation) reconstruction and is more immune to the inconsistent data problem induced by cardiac motions. The temporal resolution of multislice helical CT with the HS algorithm is comparable to that of single-slice helical CT with the HS algorithm. In practice, the 180 degrees LI and HS-LI algorithms can be used in parallel to generate two image sets from the same scan acquisition, one (180 degrees LI) for improved z-resolution and noises, and the other (HS-LI) for improved image temporal resolution. PMID- 11021683 TI - Enhanced 3-D-reconstruction algorithm for C-arm systems suitable for interventional procedures. AB - Increasingly, three-dimensional (3-D) imaging technologies are used in medical diagnosis, for therapy planning, and during interventional procedures. We describe the possibilities of fast 3-D-reconstruction of high-contrast objects with high spatial resolution from only a small series of two-dimensional (2-D) planar radiographs. The special problems arising from the intended use of an open, mechanically unstable C-arm system are discussed. For the description of the irregular sampling geometry, homogeneous coordinates are used thoroughly. The well-known Feldkamp algorithm is modified to incorporate corresponding projection matrices without any decomposition into intrinsic and extrinsic parameters. Some approximations to speed up the whole reconstruction procedure and the tradeoff between image quality and computation time are also considered. Using standard hardware the reconstruction of a 256(3) cube is now possible within a few minutes, a time that is acceptable during interventions. Examples for cranial vessel imaging from some clinical test installations will be shown as well as promising results for bone imaging with a laboratory C-arm system. PMID- 11021684 TI - Three-dimensional attenuation map reconstruction using geometrical models and free-form deformations. AB - We address the issue of using deformable models to reconstruct an unknown attenuation map of the torso from a set of transmission scans. We assume the three-dimensional (3-D) distribution of attenuation coefficients to be piecewise uniform. We represent the unknown distribution by a set of closed surfaces defining regions having the same attenuating properties. The methods of reconstruction published so far tend to directly deform the surfaces, the parameters being the surface elements. Rather than deforming the surfaces, we explore the possibility of deforming the space in which the geometrical primitives are contained. We focus on the use of free-form deformations (FFD's) to describe the continuous transformation of space used to match a set of transmission measurements. We illustrate this approach by reconstructing realistically simulated transmission scans of the torso with various noise levels and compare the results to standard reconstruction methods. PMID- 11021685 TI - Reconstruction of two- and three-dimensional images from synthetic-collimator data. AB - A novel SPECT collimation method, termed the synthetic collimator, is proposed. The synthetic collimator employs a multiple-pinhole aperture and a high resolution detector. The problem of multiplexing, normally associated with multiple pinholes, is reduced by obtaining projections at a number of pinhole detector distances. Projections with little multiplexing are collected at small pinhole-detector distances and high-resolution projections are collected at greater pinhole-detector distances. These projections are then reconstructed using the ML-EM algorithm. It is demonstrated through computer simulations that the synthetic collimator has superior resolution properties to a high-resolution parallel-beam (HRPB) collimator and a specially built ultra-high-resolution parallel-beam (UHRPB) collimator designed for our 0.38-mm pixel CdZnTe detectors. It is also shown that reconstructing images in three dimensions is superior to reconstructing them in two dimensions. The advantages of a high-resolution synthetic collimator over the parallel-hole collimators are apparently reduced in the presence of statistical noise. However, a high-sensitivity synthetic collimator was designed which again shows superior properties to the parallel hole collimators. Finally, it is demonstrated that, for the cases studied, high resolution detectors are necessary for the proper functionality of the synthetic collimator. PMID- 11021686 TI - Improved image quality and computation reduction in 4-D reconstruction of cardiac gated SPECT images. AB - Spatiotemporal reconstruction of cardiac-gated SPECT images permits us to obtain valuable information related to cardiac function. However, the task of reconstructing this four-dimensional (4-D) data set is computation intensive. Typically, these studies are reconstructed frame-by-frame: a nonoptimal approach because temporal correlations in the signal are not accounted for. In this work, we show that the compression and signal decorrelation properties of the Karhunen Loeve (KL) transform may be used to greatly simplify the spatiotemporal reconstruction problem. The gated projections are first KL transformed in the temporal direction. This results in a sequence of KL-transformed projection images for which the signal components are uncorrelated along the time axis. As a result, the 4-D reconstruction task is simplified to a series of three dimensional (3-D) reconstructions in the KL domain. The reconstructed KL components are subsequently inverse KL transformed to obtain the entire spatiotemporal reconstruction set. Our simulation and clinical results indicate that KL processing provides image sequences that are less noisy than are conventional frame-by-frame reconstructions. Additionally, by discarding high order KL components that are dominated by noise, we can achieve savings in computation time because fewer reconstructions are needed in comparison to conventional frame-by-frame reconstructions. PMID- 11021687 TI - Direct least-squares estimation of spatiotemporal distributions from dynamic SPECT projections using a spatial segmentation and temporal B-splines. AB - Artifacts can result when reconstructing a dynamic image sequence from inconsistent, as well as insufficient and truncated, cone beam single photon emission computed tomography (SPECT) projection data acquired by a slowly rotating gantry. The artifacts can lead to biases in kinetic model parameters estimated from time-activity curves generated by overlaying volumes of interest on the images. However, the biases in time-activity curve estimates and subsequent kinetic parameter estimates can be reduced significantly by first modeling the spatial and temporal distribution of the radiopharmaceutical throughout the projected field of view, and then estimating the time-activity curves directly from the projections. This approach is potentially useful for clinical SPECT studies involving slowly rotating gantries, particularly those using a single-detector system or body contouring orbits with a multidetector system. We have implemented computationally efficient methods for fully four dimensional (4-D) direct estimation of spatiotemporal distributions from dynamic SPECT projection data. Temporal B-splines providing various orders of temporal continuity, as well as various time samplings, were used to model the time activity curves for segmented blood pool and tissue volumes in simulated cone beam and parallel beam cardiac data acquisitions. Least-squares estimates of time activity curves were obtained quickly using a workstation. Given faithful spatial modeling, accurate curve estimates were obtained using cubic, quadratic, or linear B-splines and a relatively rapid time sampling during initial tracer uptake. From these curves, kinetic parameters were estimated accurately for noiseless data and with some bias for noisy data. A preliminary study of spatial segmentation errors showed that spatial model mismatch adversely affected quantitative accuracy, but also resulted in structured errors (projected model versus raw data) that were easily detected in our simulations. This suggests iterative refinement of the spatial model to reduce structured errors as an area of future research. PMID- 11021688 TI - Reconstruction of attenuation map using discrete consistency conditions. AB - Methods of quantitative emission computed tomography require compensation for linear photon attenuation. A current trend in single-photon emission computed tomography (SPECT) and positron emission tomography (PET) is to employ transmission scanning to reconstruct the attenuation map. Such an approach, however, considerably complicates both the scanner design and the data acquisition protocol. A dramatic simplification could be made if the attenuation map could be obtained directly from the emission projections, without the use of a transmission scan. This can be done by applying the consistency conditions that enable us to identify the operator of the problem and, thus, to reconstruct the attenuation map. In this paper, we propose a new approach based on the discrete consistency conditions. One of the main advantages of the suggested method over previously used continuous conditions is that it can easily be applied in various scanning configurations, including fully three-dimensional (3-D) data acquisition protocols. Also, it provides a stable numerical implementation, allowing us to avoid the crosstalk between the attenuation map and the source function. A computationally efficient algorithm is implemented by using the QR and Cholesky decompositions. Application of the algorithm to computer-generated and experimentally measured SPECT data is considered. PMID- 11021689 TI - LROC analysis of detector-response compensation in SPECT. AB - Localization ROC (LROC) observer studies examined whether detector response compensation (DRC) in ordered-subset, expectation-maximization (OSEM) reconstructions helps in the detection and localization of hot tumors. Simulated gallium (Ga-67) images of the thoracic region were used in the study. The projection data modeled the acquisition of attenuated 93- and 185-keV photons with a medium-energy parallel-hole collimator, but scatter was not modeled. Images were reconstructed with five strategies: 1) OSEM with no DRC; 2) OSEM preceded by restoration filtering; 3) OSEM with iterative DRC; 4) OSEM with an ideal DRC; and 5) filtered backprojection (FBP) with no DRC. All strategies included attenuation correction. There were four LROC studies conducted. In a study using a single tumor activity, the ideal DRC offered the best performance, followed by iterative DRC, restoration filtering, OSEM with no DRC, and FBP. Statistical significance at the 5% level was found between all pairs of strategies except for restoration filtering and OSEM with no DRC. A similar ranking was found for a more realistic study using multiple tumor activities. Additional studies considered the effects of OSEM iteration number and tumor activity on the detection improvement that iterative DRC offered with respect to OSEM with no DRC. PMID- 11021690 TI - 2.5-D simultaneous multislice reconstruction by series expansion methods from Fourier-rebinned PET data. AB - True three-dimensional (3-D) volume reconstruction from fully 3-D data in positron emission tomography (PET) has only a limited clinical use because of its large computational burden. Fourier rebinning (FORE) of the fully 3-D data into a set of 2-D sinogram data decomposes the 3-D reconstruction process into multiple 2-D reconstructions of decoupled 2-D image slices, thus substantially decreasing the computational burden even in the case when the 2-D reconstructions are performed by an iterative reconstruction algorithm. On the other hand, the approximations involved in the rebinning combined with the decoupling of the image slices cause a certain reduction of image quality, especially when the signal-to-noise ratio of the data is low. We propose a 2.5-D Simultaneous Multislice Reconstruction approach, based on the series expansion principle, where the volume is represented by the superposition of 3-D spherically symmetric bell-shaped basis functions. It takes advantage of the time reduction due to the use of the FORE (2-D) data, instead of the original fully 3-D data, but at the same time uses a 3-D iterative reconstruction approach with 3-D basis functions. The same general approach can be applied to any reconstruction algorithm belonging to the class of series expansion methods (iterative or noniterative) using 3-D basis functions that span multiple slices, and can be used for any multislice sinogram or list mode data whether obtained by a special rebinning scheme or acquired directly by a PET scanner in the 2-D mode using septa. Our studies confirm that the proposed 2.5-D approach provides a considerable improvement in reconstruction quality, as compared to the standard 2-D reconstruction approach, while the reconstruction time is of the same order as that of the 2-D approach and is clinically practical even on a general-purpose computer. PMID- 11021691 TI - Iterative crystal efficiency calculation in fully 3-D PET. AB - The calculation of the intrinsic efficiency of individual crystals is one of the steps needed to obtain accurate images of the radioisotope distribution in positron emission tomography (PET). These efficiencies can be computed by comparing the number of coincidence counts obtained when the crystals are equally illuminated by the same source. However, because the number of coincidence counts acquired for one crystal also depends on the efficiency of the other crystals in coincidence, most methods of crystal efficiency calculation need to assume that the influence of the other crystals is negligible. If there are large crystal efficiency variations, this approximation may lead to systematic errors. We have recently implemented an iterative method for a single ring of detectors that does not rely on this assumption. In this paper, we describe a fully three-dimensional (3-D) iterative method that better exploits the sensitivity of the tomograph and allows reduced acquisition times or the use of narrow energy windows. We compare the performance of the iterative method (single-ring and extended to fully 3-D) with noniterative techniques for different acquisition times of a uniform cylinder. Two different energy windows were used to assess the performance of each method with different levels of variations of crystal efficiency. The results showed that the iterative methods are more accurate when large efficiency variations exist and that only the fully 3-D methods provided good efficiency estimates with very low duration scans. We, thus, conclude that iterative fully 3 D methods provide the best estimations and can be used in a larger range of situations than can the other methods tested. PMID- 11021692 TI - Resolution and noise properties of MAP reconstruction for fully 3-D PET. AB - We derive approximate analytical expressions for the local impulse response and covariance of images reconstructed from fully three-dimensional (3-D) positron emission tomography (PET) data using maximum a posteriori (MAP) estimation. These expressions explicitly account for the spatially variant detector response and sensitivity of a 3-D tomograph. The resulting spatially variant impulse response and covariance are computed using 3-D Fourier transforms. A truncated Gaussian distribution is used to account for the effect on the variance of the nonnegativity constraint used in MAP reconstruction. Using Monte Carlo simulations and phantom data from the microPET small animal scanner, we show that the approximations provide reasonably accurate estimates of contrast recovery and covariance of MAP reconstruction for priors with quadratic energy functions. We also describe how these analytical results can be used to achieve near-uniform contrast recovery throughout the reconstructed volume. PMID- 11021693 TI - Comparison of 3-D maximum a posteriori and filtered backprojection algorithms for high-resolution animal imaging with microPET. AB - We have evaluated the performance of two three-dimensional (3-D) reconstruction algorithms with data acquired from microPET, a high resolution tomograph dedicated to small animal imaging. The first was a linear filtered-backprojection algorithm (FBP) with reprojection of the missing data, and the second was a statistical maximum a posteriori probability algorithm (MAP). The two algorithms were evaluated in terms of their resolution performance, both in phantoms and in vivo. Sixty independent realizations of a phantom simulating the brain of a baby monkey were acquired, each containing three million counts. Each of these realizations was reconstructed independently with both algorithms. The ensemble of the 60 reconstructed realizations was used to estimate the standard deviation as a measure of the noise for each reconstruction algorithm. More detail was recovered in the MAP reconstruction without an increase in noise relative to FBP. Studies in a simple cylindrical compartment phantom demonstrated improved recovery of known activity ratios with MAP. Finally, in vivo studies also demonstrated a clear improvement in spatial resolution using the MAP algorithm. The quantitative accuracy of the MAP reconstruction was also evaluated by comparison with autoradiography and direct well counting of tissue samples and was shown to be superior. PMID- 11021694 TI - Energy-based scatter correction for 3-D PET scanners using NaI(T1) detectors. AB - Earlier investigations with BGO positron emission tomography (PET) scanners showed that the scatter correction technique based on multiple acquisitions with different energy windows are problematic to implement because of the poor energy resolution of BGO (22%), particularly for whole-body studies. We believe that these methods are likely to work better with NaI(TI) because of the better energy resolution achievable with NaI(TI) detectors (10%). Therefore, we investigate two different choices for the energy window, a low-energy window (LEW) on the Compton spectrum at 400-450 keV, and a high-energy window (HEW) within the photopeak (lower threshold above 511 keV). The results obtained for our three-dimensional (3-D) (septa-less) whole-body scanners [axial field of view (FOV) of 12.8 cm and 25.6 cm] as well as for our 3-D brain scanner (axial FOV of 25.6 cm) show an accurate prediction of the scatter distribution for the estimation of trues method (ETM) using a HEW, leading to a significant reduction of the scatter contamination. The dual-energy window (DEW) technique using a LEW is shown to be intrinsically wrong; in particular, it fails for line source and bar phantom measurements. However, the method is able to produce good results for homogeneous activity distributions. Both methods are easy to implement, are fast, have a low noise propagation, and will be applicable to other PET scanners with good energy resolution and stability, such as hybrid NaI(TI) PET/SPECT dual-head cameras and future PET cameras with GSO or LSO scintillators. PMID- 11021695 TI - Evaluating performance of reconstruction algorithms for 3-D [15O] water PET using subtraction analysis. AB - Positron emission tomography (PET) [15O] activation studies have benefited significantly from three-dimensional (3-D) data acquisition. However, they have been slow to take advantage of new 3-D reconstruction techniques. Compared with the widely used 3-D reprojection reconstruction (3DRP), the advantage of signal and noise for iterative algorithms has been outweighed by concern about long and complicated reconstruction procedures and inconsistent performance. Most pseudo-3 D algorithms, such as rebinning methods, aim at increasing the speed of reconstruction but lack further resolution improvement or noise control. Although many evaluations have been conducted through simulations and phantom experiments, the spatially varying nature of signal and noise and the complexity of biological effects have complicated the interpretation of real data based on simulation or phantom results. We have taken a different approach and used the analysis of real data directly as a measure with which to compare three reconstruction algorithms: 3DRP, iterative filtered backprojection with median root prior (IFBP-MRP), and Fourier rebinning followed by two-dimensional (2-D) filtered backprojection (FORE FBP) for [15O] PET. Two subjects, each with 32 scans acquired in four sessions during a finger opposition motor task, are analyzed using subtraction. A fixed volume-of-interest (VOI) measurement in regions related to the task demonstrates that at high resolution, IFBP-MRP has the best signal-to-noise performance followed by 3DRP and FORE-FBP; however, this advantage gradually diminishes as the resolution decreases. For a voxel measurement derived from the image of each reconstruction, all three algorithms are capable of detecting highly activated regions. Although there are some differences in the size, shape, and center location of the activated foci, our preliminary results suggest that IFBP-MRP does offer enhanced signal with some noise control compared with 3DRP for the analysis of high-resolution images. If images are to be analyzed at an intermediate to lower resolution, FORE-FBP provides a significant reduction of reconstruction time compared with 3DRP. PMID- 11021696 TI - List-mode maximum-likelihood reconstruction applied to positron emission mammography (PEM) with irregular sampling. AB - We present a preliminary study of list-mode likelihood reconstruction of images for a rectangular positron emission tomograph (PET) specifically designed to image the human breast. The prospective device consists of small arrays of scintillation crystals for which depth of interaction is estimated. Except in very rare instances, the number of annihilation events detected is expected to be far less than the number of distinguishable events. If one were to histogram the acquired data, most histogram bins would remain vacant. Therefore, it seems natural to investigate the efficacy of processing events one at a time rather than processing the data in histogram format. From a reconstruction perspective, the new tomograph presents a challenge in that the rectangular geometry leads to irregular radial and angular sampling, and the field of view extends completely to the detector faces. Simulations are presented that indicate that the proposed tomograph can detect 8-mm-diameter spherical tumors with a tumor-to-background tracer density ratio of 3:1 using realistic image acquisition parameters. Spherical tumors of 4-mm diameter are near the limit of detectability with the image acquisition parameters used. Expressions are presented to estimate the loss of image contrast due to Compton scattering. PMID- 11021697 TI - A 3-D reconstruction system for the human jaw using a sequence of optical images. AB - This paper presents a model-based vision system for dentistry that will assist in diagnosis, treatment planning, and surgical simulation. Dentistry requires an accurate three-dimensional (3-D) representation of the teeth and jaws for diagnostic and treatment purposes. The proposed integrated computer vision system constructs a 3-D model of the patient's dental occlusion using an intraoral video camera. A modified shape from shading (SFS) technique, using perspective projection and camera calibration, extracts the 3-D information from a sequence of two-dimensional (2-D) images of the jaw. Data fusion of range data and 3-D registration techniques develop the complete jaw model. Triangulation is then performed, and a solid 3-D model is reconstructed. The system performance is investigated using ground truth data, and the results show acceptable reconstruction accuracy. PMID- 11021698 TI - Unmatched projector/backprojector pairs in an iterative reconstruction algorithm. AB - Computational burden is a major concern when an iterative algorithm is used to reconstruct a three-dimensional (3-D) image with attenuation, detector response, and scatter corrections. Most of the computation time is spent executing the projector and backprojector of an iterative algorithm. Usually, the projector and the backprojector are transposed operators of each other. The projector should model the imaging geometry and physics as accurately as possible. Some researchers have used backprojectors that are computationally less expensive than the projectors to reduce computation time. This paper points out that valid backprojectors should satisfy a condition that the projector/backprojector matrix must not contain negative eigen-values. This paper also investigates the effects when unmatched projector/backprojector pairs are used. PMID- 11021699 TI - Analytical versus voxelized phantom representation for Monte Carlo simulation in radiological imaging. AB - Monte Carlo simulations in nuclear medicine, with accurately modeled photon transport and high-quality random number generators, require precisely defined and often detailed phantoms as an important component in the simulation process. Contemporary simulation models predominantly employ voxel-driven algorithms, but analytical models offer important advantages. We discuss the implementation of ray-solid intersection algorithms in analytical superquadric-based complex phantoms with additional speed-up rejection testing for use in nuclear medicine imaging simulations, and we make comparisons with voxelized counterparts. Comparisons are made with well-known cold rod:sphere and anthropomorphic phantoms. For these complex phantoms, the analytical phantom representations are nominally several orders of magnitude smaller in memory requirements than are voxelized versions. Analytical phantoms facilitate constant distribution parameters. As a consequence of discretizing a continuous surface into finite bins, for example, time-dependent voxelized phantoms can have difficulties preserving accurate volumes of a beating heart. Although virtually no inaccuracy is associated with path calculations in analytical phantoms, the discretization can negatively impact the simulation process and results. Discretization errors are apparent in reconstructed images of cold rod:sphere voxel-based phantoms because of a redistribution of the count densities in the simulated objects. These problems are entirely avoided in analytical phantoms. Voxelized phantoms can accurately model detailed human shapes based on segmented computed tomography (CT) or magnetic resonance imaging (MRI) images, but analytical phantoms offer advantages in time and accuracy for evaluation and investigation of imaging physics and reconstruction algorithms in a straightforward and efficient manner. PMID- 11021700 TI - Ethical standards in the insurance industry. PMID- 11021701 TI - Osteoradionecrosis of the jaws: clinical characteristics and relation to the field of irradiation. AB - PURPOSE: This study focuses on the clinical characteristics of patients with osteoradionecrosis (ORN) of the jaws and on the relation between the extent of the ORN and the field of irradiation. PATIENTS AND METHODS: The study group consisted of 80 patients referred for treatment of ORN. Charts of the extent of ORN, based on panoramic radiographs, were drawn by the clinical investigator and compared with charts of the field of irradiation drawn by the oncologists. Also recorded was diagnosis of the lesion, stage, location, treatment schedule, and period and dose of irradiation. Also documented were various potential initiating factors for ORN. RESULTS: Smoking habits of ORN patients were similar to those of other patients with head and neck cancer. A new primary tumor or a recurrence was diagnosed in 10% of the patients at the time of ORN. Only 3 patients had received accumulated doses of less than 60 Gy. More than half of the cases were initiated by removal of teeth; however, one third occurred spontaneously. ORN developed within the first 3 years in 74%; but ORN can emerge on a traumatic basis for an infinite number of years after radiation therapy. Widespread ORN may be symptomless apart from a discrete dehiscense of the oral mucosa. The predilection site for ORN is the mandibular molar region. All cases of ORN but 1 were found in the field of radiation. CONCLUSION: Because many ORN case are symptomless, more focus on mucosal dehiscence in the follow-up after radiation therapy is advocated in an effort to detect ORN at an early stage. The consequence of practically all ORN cases being located in the field of irradiation, together with tooth removal frequently being the initiating factor, should lead to a more aggressive preirradiation approach to dental pathology located within the field of radiation. PMID- 11021702 TI - The minor oral surgery outcome scale: a multi-attribute patient-derived outcome measure. AB - PURPOSE: The purpose of this study was to produce a clinically useful, patient derived, minor oral surgery outcome scale. PATIENTS AND METHODS: Seventy-seven consecutive patients scheduled for minor oral surgery were interviewed using semistructured interview techniques to identify those areas of life (domains) that patients believed will be affected by minor oral surgery. These interviews were analyzed by a multidisciplinary panel and a scale based on 5 domains, each with 4 outcome statements was constructed. The domains and outcome statements were weighted in terms of importance by a further consecutive sample of 100 patients using resource allocation and visual analog tasks. An additive mathematical formula was applied to the mean weights for each domain and outcome states to produce the final weighted scale. RESULTS: Five domains were identified from the interviews and weighted according to their order of importance (0 to 100 scale; 0 = least important). These were general health and well-being (24.6); impact on home/social life (20.8); health and comfort of the mouth, teeth, and gums (20.0); appearance (18.8); and impact on job/studies (15.8). CONCLUSION: A clinically applicable, multi-attribute outcome scale has been produced that takes into account all determinants of health outcome as perceived by the patient in the context of minor oral surgery. It is completed by selecting a weighted statement from each domain, the sum of which constitutes the patient's health state utility score. The scale has potentially extensive application to both clinical care and research. PMID- 11021703 TI - Nasolabial flap reconstruction of oral cavity defects: a report of 18 cases. AB - PURPOSE: This article describes the successful use of pedicled nasolabial flaps in the reconstruction of various oral cavity defects with or without adjunctive microvascular free tissue transfer. PATIENTS AND METHODS: Twenty-eight flaps were performed in 18 patients, for a variety of oral cavity defects. RESULTS: All flaps healed without evidence of necrosis, infection, or dehiscence. Patient satisfaction with this procedure was high. The use of the nasolabial flap appears to provide an improvement in overall functional outcome. CONCLUSIONS: The inferiorly based nasolabial flap provides reliable coverage of intermediate-sized oral cavity defects when used alone. It can improve mastication and speech when used in conjunction with microvascular free tissue transfer for the reconstruction of large combined defects of the tongue and floor of mouth. PMID- 11021704 TI - Mandibular fracture after third molar removal. AB - PURPOSE: Mandibular fracture after impacted lower third molar removal is a rare, but major, complication. The factors leading to a mandibular fracture secondary to third molar removal are analyzed retrospectively. PATIENTS AND METHODS: Six patients who suffered from a mandibular fracture as a complication after third molar removal were examined clinically and radiographically. RESULTS: All fractures occurred an average of 14 days postoperatively. The patients were 42 to 50 years old and were all fully dentulous. All grades of tooth impaction were included. CONCLUSIONS: The major risk factor for this complication seems to be advanced age in combination with a full dentition. The degree of tooth impaction is less important. Preexisting bone lesions weaken the mandible and further predispose to a fracture. PMID- 11021705 TI - Benign tumors of adipose tissue of the oral cavity: a clinicopathologic study of 13 cases. AB - PURPOSE: The purpose of this study was to determine the clinical and histologic features of 9 simple lipomas, 2 fibrolipomas, and 2 infiltrating lipomas of the oral cavity. In addition, 10 cases of infiltrating lipoma of the oral cavity previously reported in the literature were reviewed. MATERIALS AND METHODS: All cases were retrieved from the archives of the Department of Oral Medicine/Pathology of the University of Thessaloniki, Greece. Data on age, sex, location, clinical appearance, duration, recurrence, and histologic appearance were evaluated. RESULTS: This study showed that the sex distribution of the simple lipomas was approximately equal, and the mean age of patients was 60.2 years. The buccal mucosa was the most frequent location of the tumors. Their size ranged from 0.2 to 1.5 cm, with an average rate 0.8 cm. In 3 cases, the tumor was not encapsulated. The sex distribution of the infiltrating lipomas was equal, and the mean age of the patients was 36.8 years. The most frequent location was the tongue. CONCLUSIONS: Tumors of adipose tissue represent rare neoplasms of the oral cavity. The diagnosis is based on both the clinical and histologic characteristics. Caution is required during their surgical excision to avoid recurrence, especially with infiltrating lipomas. PMID- 11021706 TI - A comparison of labial and crestal incisions for the 1-stage placement of IMZ implants: a pilot study. AB - PURPOSE: The aim of this study was to compare the crestal incision with the labial flap design when inserting a 2-stage implant system in a nonsubmerged manner. PATIENTS AND METHODS: Ten consecutive edentulous patients with a severely resorbed mandible (Cawood Class V to VI) that resulted in reduced stability and insufficient retention of the lower denture were included. In all patients, 2 IMZ implants were inserted in a 1-stage procedure in the mandibular canine regions as part of an implant overdenture treatment. In 5 patients, the labial flap approach was used, and a crestal incision approach was used in the other 5 patients. Standardized evaluations were performed at 2, 6, and 12 weeks after implant placement and 12 months after placement of the new prosthesis. RESULTS: In the first postoperative weeks, more hyperplasia occurred around the implants inserted by the crestal incision than the labial flap. However, after 1 year of function, no striking differences between the 2 groups were present with regard to the clinical and radiographic parameters. CONCLUSIONS: From this pilot study, it was concluded that both the crestal incision and the labial flap approach are reliable procedures for insertion of IMZ implants in a 1-stage procedure. However, because of the smaller risk of soft tissue overgrowth, there is a preference for the labial flap approach. PMID- 11021707 TI - The presence of collagen III and IV in experimental microsurgical anastomoses of arteries. AB - PURPOSE: This study investigated the distribution of collagen III and IV in experimental microvascular anastomoses and correlated the findings with the quality of the healing process. MATERIALS AND METHODS: A previously excised and reanastomosed section of the left common carotid artery of 21 female Wistar rats were explanted 4 weeks after surgery. Collagens III and IV were identified in perfusion-fixed specimens by immunohistochemistry. RESULTS: The identification and distribution of collagens reflected the quality of each suturing procedure. A myointimal hyperplasia was present, extending into the vascular lumen, and was closely related to the ends of the vessel segments. The extent of myointimal hyperplasia was dependent on the length and the vitality of the vessel segments. The media was the weakest part of the tissue layers with regard to regenerative potential in the axial direction. The marked cuff of the adventitia around the anastomosis was rich in collagen III-positive vasa vasorum. Collagen IV was distributed in net-like patterns in the media. The distribution pattern in the media was not observed in anastomoses with incomplete approximation and necrotic segment ends. The collagen IV-positive subendothelial basal membrane was incomplete in the aneurysms because of partial necrosis. CONCLUSIONS: The experimental findings support the need for atraumatic suturing in microvascular surgery. Immunohistochemical detection of collagens can be a valuable tool for studying the basic processes of wound healing and the integration of microvascular flaps into their recipient sites. PMID- 11021708 TI - Presence of human papilloma viruses in intraosseous ameloblastoma. AB - PURPOSE: This study investigated the possibility that human papilloma virus (HPV) is a possible etiologic agent in the development of ameloblastoma. MATERIALS AND METHODS: DNA was extracted from 18 formalin-fixed, paraffin-embedded biopsy specimens and assayed for the presence of HPV DNA by PCR using the L1 consensus primer and specific primers for HPV types 6/11, 16 and 18. RESULTS: Eight samples (67%) were positive for HPV. Of the 8 HPV-positive samples, 7 were positive for HPV 18. Four of the HPV 18-positive samples were also positive for HPV 6/11. One HPV-positive sample was not positive for any of the type-specific primers. CONCLUSIONS: No conclusions can be drawn about the etiologic role of HPV from this study, but surgical manipulation is suggested to be one of the reasons for HPV presence attributable to contamination from the surface mucosal epithelium in these tumors. PMID- 11021709 TI - Reactive arthritis: preliminary microbiologic analysis of the human temporomandibular joint. AB - PURPOSE: The presence of Chlamydia trachomatis has been previously shown in the temporomandibular joint (TMJ). This study investigated whether the presence of other bacteria associated with reactive arthritis (ReA) can be identified in the TMJ. MATERIALS AND METHODS: Posterior bilaminar tissue removed during TMJ surgery from 26 patients (24 F, 2 M) was evaluated for the presence of C. trachomatis, Mycoplasma fermentans, Mycoplasma genitalium, Campylobacter jejuni, Yersinia enterocolitica, Salmonella spp, and Shigella spp by highly specific polymerase chain reaction (PCR) assays. RESULTS: Bacterial DNA was identified in the TMJ as follows: C. trachomatis, 11 of 26 (42%); M. fermentans/orale, 6 of 26 (23%); M. genitalium, 9 of 26 (35%). Nine of 26 TMJs (35%) had the presence of a single bacterial species. Eight of 26 TMJs (31%) had more than 1 species, as follows: C. trachomatis with either M. genitalium or M. fermentans/orale in 5 of 26 (19%), M. fermentans/orale with M. genitalium 2 of 26 (8%), and C. trachomatis/M. fermentans/orale/M. genitalium, 1 of 26 (4%). A total of 17 of 26 (65%) of TMJs had the presence of bacteria identified in the TMJ. Campylobacter jejuni, Y. enterocolitica, Salmonella spp, and Shigella spp were not identified in any samples. CONCLUSIONS: The presence of M. genitalium in the human TMJ has not been previously reported. The presence of bacteria in the TMJ, either singly or concurrently with other bacteria, may serve as the pathogenetic mechanism of TMJ inflammation. The presence of 2 bacteria from the urogenital tract in the TMJ suggests that internal derangement of the TMJ may occur as a result of a sexually acquired infection. PMID- 11021710 TI - Persistent anterior maxillary bone loss. PMID- 11021711 TI - Peripheral and mural ameloblastoma in the mandibular canine region of a 13-year old boy. PMID- 11021712 TI - Mandibular metastasis of bladder transitional cell carcinoma. PMID- 11021713 TI - Epineural dissection to preserve the inferior alveolar nerve in excision of an ameloblastoma of mandible: case report. PMID- 11021714 TI - Adenomatoid odontogenic tumor: a case report. PMID- 11021715 TI - Spindle cell squamous carcinoma of the maxilla: report of a case with immunohistochemical analysis. PMID- 11021716 TI - Congenital dilation of the submandibular duct: report of a case. PMID- 11021717 TI - Schwannoma of the upper lip: ultrasound, CT, and MRI findings. PMID- 11021718 TI - Cystic lymphoid hyperplasia in the parotid gland in HIV infection: report of 2 cases. PMID- 11021719 TI - Body dysmorphic disorder involving the facial region: a report of 6 cases. PMID- 11021720 TI - Giant parotid pleomorphic adenoma involving the parapharyngeal space: report of a case. PMID- 11021721 TI - A method for reduction of zygomaticomaxillary complex fractures. PMID- 11021722 TI - A modification to the mucosal closure after alveolar cleft bone grafting. PMID- 11021723 TI - A discussion about the discussion. Nasal profile changes after maxillary impaction and advancement surgery. PMID- 11021724 TI - Practice transitions can be difficult. PMID- 11021725 TI - Oral cancer information too difficult for most, study says. PMID- 11021726 TI - Patient pain and anxiety: the medical and psychologic challenges facing oral and maxillofacial surgery. PMID- 11021727 TI - Guidelines for anxiety control and pain management in oral and maxillofacial surgery. PMID- 11021728 TI - Nonsteroidal anti-inflammatory drugs and opioids: safety and usage concerns in the differential treatment of postoperative orofacial pain. PMID- 11021729 TI - Pain management in the multiply operated temporomandibular joint patient. PMID- 11021730 TI - An open-label evaluation of the efficacy and safety of Stadol NS with ibuprofen in the treatment of pain after removal of impacted wisdom teeth. AB - PURPOSE: This study evaluated the efficacy and safety of transnasal butorphanol tartrate (Stadol NS; Bristol-Myers Squibb, New York, NY) in the treatment of patients with moderate to severe pain after oral surgery for the removal of impacted third molars. PATIENTS AND METHODS: In this open-label study, third molar extraction was performed on 25 male and 25 female patients. These patients were given Stadol NS nasal spray, 1 mg, administered in a single dose every 4 hours as needed. However, patients were allowed to remedicate 60 to 90 minutes after the initial dose, if necessary. They also took ibuprofen (400 mg) as concomitant medication every 4 to 6 hours for the first 48 hours. Patients recorded pain intensity on a visual analog scale, with 0 representing no pain, to 10 representing the most severe pain. RESULTS: Stadol NS significantly reduced pain (by 50%) after third molar extraction within 15 minutes after administration. It had a high level of patient acceptance, 81% on day 1, and was well tolerated. CONCLUSION: The rapid onset of analgesia and long duration of action shown by Stadol NS in this study, as well as its ease of administration and high level of patient acceptance, suggest that this drug would be an excellent primary choice for the management of pain after third molar extraction and oral surgery in general. PMID- 11021731 TI - Analgesic efficacy of intranasal butorphanol (Stadol NS) in the treatment of pain after dental impaction surgery. AB - PURPOSE: This study evaluated the safety and efficacy of increasing doses of intranasal butorphanol (Stadol NS, Bristol-Myers Squibb, New York, NY) compared with placebo in controlling moderate to severe pain after removal of bony impacted third molars. PATIENTS AND METHODS: This single-dose, double-blind, parallel-group, dose-response trial compared the efficacy and safety of 4 doses of intranasally administered butorphanol tartrate and placebo in controlling moderate to severe pain after the removal of impacted third molars in 151 patients. The study was conducted at 2 sites. The patients were randomly assigned to receive 1 dose of butorphanol tartrate: 0.25 mg (n = 31), 0.5 mg (n = 29), 1.0 mg (n = 30), 2.0 mg (n = 30), or placebo (n = 31). Medication was administered with a metered-dose spray pump. Patients rated pain intensity (PI), pain relief (PAR), pain half gone (PHG), and adverse events at 0.25, 0.5, 1, 2, 3, 4, 5, and 6 hours after treatment. At the end of the study period or before rescue medication (ibuprofen, 400 mg, or acetaminophen, 1,000 mg), patients provided an overall assessment (GLOBAL). RESULTS: A linear dose-response regression (P < or = .05) was observed for the means of pain intensity difference (PID), PAR, and PHG at 0.25, 0.5, and 1 hour, and for sum of pain intensity differences (SPID), sum of pain relief (TOTPAR), peak PID and PAR, and GLOBAL evaluation. The 1.0- and 2.0-mg groups experienced greater pain relief compared with placebo (P = .05) during the first hour after drug administration. The 1.0- and 2.0-mg groups had significantly better GLOBAL evaluations than the placebo group, but were not significantly different from placebo for time until remedication (TREMED). Incidence and severity of the most common adverse events were dose-related. Two severe adverse events (drowsiness and dizziness) occurred after the 2.0-mg dose. CONCLUSION: Intranasal butorphanol effectively relieved postsurgical dental pain, with a rapid onset within 15 minutes, and seems to be a promising addition to the current armamentarium of opioid analgesics. As with other opioids, it should be used cautiously in an outpatient setting. PMID- 11021732 TI - Effectiveness of Stadol NS (butorphanol tartrate) with ibuprofen in the treatment of pain after laser-assisted uvulopalatopharyngoplasty. AB - PURPOSE: Laser-assisted uvulopalatopharyngoplasty (LA-UPPP) is a recently developed technique for treating snoring and mild sleep apnea. This treatment is associated with severe postoperative pain. This study evaluated the effectiveness and safety of transnasal administration of butorphanol tartrate (Stadol NS; Bristol-Myers Squibb, New York, NY) as a primary means of controlling pain after this surgical procedure. PATIENTS AND METHODS: Twenty patients (14 men and 6 women; mean age, 43 years) were enrolled and treated with LA-UPPP and radiofrequency turbinate procedures (coblation) for severe snoring and chronic nasal congestion in an open label study. The same surgeon performed all procedures. Patients were advised to take 2 tablets of ibuprofen (200 mg each) and use a single 1 mg spray of Stadol NS in 1 nostril for the initial dose. If adequate pain relief was not achieved in 1 to 1.5 hours, an additional 1 mg dose of Stadol NS was recommended. Thereafter, they could repeat the ibuprofen and Stadol NS every 3 to 4 hours, as needed. Comprehensive preoperative and postoperative evaluations were made using a visual analog scale of 0 to 10, with 0 equal to no pain and 10 being the most severe. Patients were followed-up for 14 days. RESULTS: All 20 patients had LA-UPPP and nasal turbinate radiofrequency coblation, and all 20 were eligible for both efficacy and safety analysis. Pain was reduced from a mean of 6.6 to a mean of 3.3 (50%) after an average of 48 minutes after the initial dose. CONCLUSION: Stadol NS appears to be an effective treatment for moderate-to-severe pain after LA-UPPP. If patients have difficulty swallowing, which is the case with most throat surgeries performed by oral and maxillofacial surgeons and otolaryngologists, the transnasal route of administration reduces the number of oral medications taken by the patient. PMID- 11021733 TI - Phase I dose escalation pharmacokinetics of O-(chloroacetylcarbamoyl) fumagillol (TNP-470) and its metabolites in AIDS patients with Kaposi's sarcoma. AB - The pharmacokinetics of TNP-470 and its major metabolites were investigated in AIDS patients enrolled in a phase I dose escalation trial for the treatment of Kaposi's sarcoma. The patients received TNP-470 by 1-h intravenous infusion in dose cohorts of 10, 20, 30, 40, 50 and 70 mg/m2. The parent drug and metabolites, MII and MIV, were measured by high-performance liquid chromatography/mass spectrometry (HPLC/MS) in plasma samples collected during and out to 168 h after the beginning of the infusion. Both metabolites were detected in all patients' plasma, while the parent drug was undetectable at time-points as early as 5 min after the end of infusion for some patients. A large interpatient variability of pharmacokinetic parameters among the dosing cohorts was observed for TNP-470, with a mean (+/- SD) plasma elimination half-life (t1/2) of 0.06 +/- 0.04 h, plasma clearance (CL) of 1487 +/- 1216 l/h and an area under the concentration versus time curve (AUC) of 49.9 +/- 35.8 ng/ml x h. Time to maximum plasma concentration (Tmax) typically occurred before the end of the infusion. The predominant plasma metabolite was MII with a t1/2 of 1.21 +/- 0.43 h, AUC of 1226 +/- 2303 l/h and a Tmax occurring between 5 and 15 min after infusion. The reported active metabolite MIV had a t1/2 of 0.24 +/- 0.13 h, AUC of 24.9 +/- 32.6 ng/ml x h and a Tmax occurring between the midpoint of the infusion and 15 min after infusion. The parent drug was undetectable by HPLC/MS/MS in urine samples collected and pooled between 0-6 and 6-24 h from the beginning of drug administration. Metabolite MIV was present in the 0-6-h urine pool of two patients enrolled in the highest dosing cohorts, equivalent to 0.4% of the administered dose. Metabolite MII was present in all 0-6-h samples analyzed and represented 1.12 +/- 0.9% of the administered dose. Renal clearance (CLR) for MII was 140 +/- 70 ml/h. PMID- 11021734 TI - Feasibility of a novel weekday-on/weekend-off oral UFT schedule as postoperative adjuvant chemotherapy for colorectal cancer. UFT Compliance Study Group, Kanagawa, Japan. AB - PURPOSE: When oral anticancer agents are used for adjuvant chemotherapy of colorectal cancer, compliance and feasibility become issues because of the long treatment time. Appropriate studies of these issues are lacking. We investigated compliance and feasibility during a weekday-on/weekend-off schedule of oral UFT (uracil-tegafur) over a period of 1 year administered as adjuvant chemotherapy to patients with colorectal cancer. PATIENTS AND METHODS: A UFT dose of 600 mg/day was prescribed according to a weekday-on/weekend-off schedule to 87 patients after potentially curative resection. Compliance was investigated in three ways: physician interview, patient self-report, and chemical analysis of urine. The results were compared with the dose prescribed. Feasibility was evaluated on the basis of two indices: relative performance (RP), which was the ratio of the actual total dose taken to the total dose planned, and individual dose intensity (IDI), which was the ratio of the actual dose taken to the dose planned during a given period. RESULTS: The compliance assessed by physician interview and by patient self-report conformed well with the prescribed dose, the rate of agreement among the three compliance measures being more than 94%. Chemical analysis of urine in 38 of the patients revealed that they were actually taking the drug. The RP was 0.72, and the IDI was 0.8. CONCLUSION: From these results, the feasibility of the weekday-on/weekend-off schedule was judged to be good. It is suggested that the feasibility would be even better if the dose of UFT was set according to body surface area. PMID- 11021735 TI - Effect of tamoxifen pretreatment on the pharmacokinetics, metabolism and cardiotoxicity of doxorubicin in female rats. AB - The purpose of this study was to examine the effect of tamoxifen pretreatment on the metabolism and pharmacokinetics of doxorubicin. We tested the hypothesis that the pretreatment would counteract the side effects of doxorubicin and modify the disposition of the drug. The concentration-time profiles of doxorubicin in plasma and blood cells were determined in conjunction with the cumulative amount of renal and hepatobiliary elimination of unchanged drug and metabolites following a 10-day tamoxifen pretreatment at a dose of 1 mg/kg per day. Furthermore, under the same experimental protocol the serum concentration-time profile of endothelin was determined as a biomarker of toxicity. METHODS: Female Sprague Dawley rats (225-275 g). pretreated orally for 10 days with corn oil or tamoxifen in corn oil (1 mg/kg per day), received 14C-doxorubicin (specific activity 0.4 microCi/mg, 10 mg/kg) intravenously. Plasma, blood cells, bile and urine were collected periodically and analyzed for doxorubicin and its metabolites. Four other groups of animals received the same pretreatment and non-labeled doxorubicin. Their serum samples were analyzed for endothelin. Two additional groups were also used to examine the effect of tamoxifen on the in vitro metabolism of doxorubicin by the cytosolic enzyme aldo-keto reductase. RESULTS: Tamoxifen pretreatment reduced the total protein of the cytosolic fraction by 50% and reduced the formation of doxorubicinol both in vitro and in vivo. The pretreatment resulted in a notable increase in the area under plasma and blood cells concentration-time curves of doxorubicin and a significant reduction in mean residence time, apparent volume of distribution and serum endothelin levels. CONCLUSIONS: We attributed the increase in the area under the curves of plasma and blood cells following tamoxifen pretreatment to a reduction in the uptake of doxorubicin by peripheral tissues. This conclusion was consistent with the reduction in the volume of distribution of plasma, mean residence time and higher availability of the parent compound for excretion. An interesting observation was that the increase in concentration of doxorubicin in plasma was not concomitant with an increase in concentration of doxorubicinol. The levels of this toxic metabolite and its corresponding biliary rate constant were reduced by approximately 50%. The results demonstrate that tamoxifen, in addition to being a modulator of P glycoprotein and counteracting the effects of doxorubicin at the cellular level, also alters the metabolic profile of doxorubicin either by inhibiting the formation of the toxic metabolite doxorubicinol or by reducing the enzyme responsible for the biotransformation. The change in metabolism may well be a contributing factor to reduction of serum endothelin levels. PMID- 11021736 TI - Ara-C differentially affects multiprotein forms of human cell DNA polymerase. AB - PURPOSE: The antimetabolite 1-beta-D-arabinofuranosylcytosine (ara-C) has proven to be one of the most effective agents available for the treatment of acute leukemia although the precise mechanism by which ara-C induces cytotoxicity remains unclear. Our laboratory has previously isolated from human cells a DNA replication complex, termed the DNA synthesome, which is fully competent to orchestrate, in vitro, all of the reactions required to efficiently and faithfully replicate DNA. Using this system and the active metabolite of ara-C, ara-CTP, we demonstrated that the human DNA synthesome can efficiently incorporate ara-CTP into internucleotide positions of newly replicated DNA in vitro mimicking results obtained using intact cells and isolated nuclei. We then hypothesized that DNA polymerase auxiliary proteins, present within the DNA synthesome, may aid in incorporating this nucleotide analog into DNA. METHODS: To test this hypothesis, we utilized three distinct multiprotein complexes each of which contained human DNA polymerase alpha and examined with standard in vitro polymerase assays the effectiveness of ara-C in inhibiting various aspects of their polymerase function. RESULTS AND CONCLUSION: These polymerase-mediated elongation assays, which included ara-CTP- or ara-C-containing primers in the reaction mixture, showed that the rate of DNA elongation in the presence of ara CTP was significantly enhanced when the DNA polymerase was associated with its auxiliary proteins, and that the elongation resulted in the formation of internucleotide ara-CMP. Nevertheless, the enhanced activities resulting from the association of these auxiliary proteins with polymerase alpha did not fully account for the remarkable efficiency with which the DNA synthesome incorporated ara-C into internucleotide positions during DNA replication. PMID- 11021737 TI - Pretreatment with ranitidine does not reduce the bioavailability of orally administered topotecan. AB - PURPOSE: The purpose of this randomized, two-period crossover study was to determine the pharmacokinetics of orally administered topotecan in the presence and absence of oral ranitidine. METHODS: Patients with solid malignant tumors refractory to standard treatment were given topotecan orally on a daily times five schedule repeated every 21 days. Topotecan was given initially at 2.3 mg/m2 per day; dose adjustments were permitted after the first dose of course 2 if necessary. Blood samples for pharmacokinetic assessments were drawn at protocol specified times for up to 10 h following oral administration of topotecan on day 1 of courses 1 and 2. Patients were randomly assigned to receive a total of nine doses of ranitidine: 150 mg twice daily for 4 days before day 1 of one of the first two courses and 150 mg given 2 h before the first topotecan dose. Plasma samples were assayed for concentrations of active topotecan lactone (TPT-L) and total topotecan (TPT-T, lactone plus open-ring carboxylate form) using high performance liquid chromatography with fluorescence detection. After completion of courses 1 and 2, patients could continue on therapy for days 1-5 of every 21 days if not withdrawn due to unacceptable toxicity, disease progression, protocol violation, or by request. Patients continued on treatment for a maximum of six courses. RESULTS: No pharmacokinetic parameter for either TPT-L or TPT-T differed significantly during administration of topotecan with ranitidine compared with topotecan alone (n = 13). Geometric mean ratios (95% confidence intervals, CIs) of areas under the curve in the presence and absence of ranitidine were 0.94 (0.80, 1.10) for TPT-L and 0.97 (0.80, 1.16) for TPT-T. Corresponding ratios (CIs) of peak plasma concentrations in the presence and absence of ranitidine were 1.06 (0.78, 1.44) for TPT-L and 1.07 (0.84, 1.38) for TPT-T. The median difference in time to peak plasma concentration was 0.0 h for TPT-L and -0.5 h for TPT-T (i.e. slightly faster in the presence of ranitidine). CONCLUSIONS: Administration of ranitidine prior to oral topotecan resulted in a similar extent of absorption. A slightly faster rate of absorption of topotecan was also observed, which is unlikely to be of clinical significance. Dosage adjustments of orally administered topotecan should not be necessary in patients who are pretreated with ranitidine, an H2 antagonist, or another agent that comparably raises gastric pH. PMID- 11021738 TI - Modified irinotecan hydrochloride (CPT-11) administration schedule improves induction of delayed-onset diarrhea in rats. AB - PURPOSE: Clinically, diarrhea is the major dose-limiting toxicity of irinotecan hydrochloride (CPT-11). Using a rat model, we attempted to decrease the incidence of delayed-onset diarrhea by modifying the administration schedule of CPT-11, and studied the pharmacokinetics in this model in relation to the incidence of diarrhea. METHODS: CPT-11 (total dose, 240 mg/kg) was administered intravenously (i.v.) to rats according to various schedules, and the incidence of delayed-onset diarrhea was monitored. RESULTS: Administration of CPT-11 at a dose of 60 mg/kg once daily for four consecutive days induced severe diarrhea, while at 30 mg/kg twice daily at an interval of 9 h (daily dose 60 mg/kg) for four consecutive days alleviated the diarrheal symptoms, and at 30 or 40 mg/kg once daily for eight or six consecutive days, respectively. diarrhea was hardly induced. With the first schedule, mucosal impairment of the cecal epithelium was observed, including wall thickening, edema, decrease in crypt number and size, and formation of pseudomembrane-like substance, whereas these changes were less severe with the second schedule and were hardly observed with the other two schedules. The areas under the plasma and cecal tissue concentration-time curves (AUCpla and AUCcec), the maximum plasma concentrations (Cmax) and the biliary excretions of CPT-11 and its metabolites, 7-ethyl-10-hydroxycamptothecin (SN-38) and SN-38 glucuronide (SN 38G) in rats depended on the daily dose of CPT-11. Exceptionally, CPT-11 Cmax was significantly lower and SN-38 AUCcec was larger in the animals treated at 30 mg/kg twice daily than in those treated at 60 mg/kg once daily. CONCLUSION: These results suggested that the duration of exposure to both CPT-11 and SN-38 of the intestinal epithelium and CPT-11 plasma Cmax are closely related to the incidence and severity of CPT-11-induced delayed-onset diarrhea in rats. PMID- 11021740 TI - Population pharmacokinetics of pemetrexed disodium (ALIMTA) in patients with cancer. AB - PURPOSE: To evaluate the population pharmacokinetics of pemetrexed disodium in cancer patients enrolled in four different open-label, multicenter, nonrandomized phase II studies. METHODS: Pemetrexed disodium was administered as a 10-min intravenous infusion (600 mg/m2) every 21 days. A total of four blood samples were to be collected each cycle per patient (n= 103 patients) during cycles 1 and 3. Plasma concentration-time data were analyzed by nonlinear mixed-effect modeling using NONMEM to estimate pemetrexed disodium pharmacokinetic parameters (mean, and between- and within-patient variability) as well as relationships between the pharmacokinetic parameters and various patient-specific factors (demographic and physiologic data). RESULTS/CONCLUSIONS: The pharmacokinetics of pemetrexed disodium were best characterized by a two-compartment model with initial distribution and terminal elimination half-lives of 0.63 h and 2.73 h, respectively. The typical value of systemic clearance (CL) in liters per hour included a relationship to creatinine clearance (CrCL) with a slope of 0.0292. Typical values of central volume (V(c)), distributional CL (Q), and peripheral volume (V(p)) were 11.3 1, 3.21 l/h, and 5.20 l, respectively. Between-patient variability was 19.6%, 15.6%, and 21.7% for CL, V(c), and V(p), respectively. A combined additive/proportional error model was used to describe residual variability, with a coefficient of variation of 23.7% for the proportional component and a standard deviation of 0.0410 microg/ml for the additive component. Significant patient-specific factors on CL were calculated CrCL, body weight, and to a lesser extent alanine transaminase and folate deficiency. Gender and body weight were significant factors on V(c) while both body surface area and albumin were significant factors on V(p). In conclusion, population pharmacokinetic modeling revealed relationships between pharmacokinetic parameters and various patient specific factors. PMID- 11021739 TI - Cisplatin versus cisplatin combined with piroxicam in a canine model of human invasive urinary bladder cancer. AB - PURPOSE: More than 12,000 people are expected to die from invasive transitional cell carcinoma (TCC) of the urinary bladder each year in the United States, indicating that more effective therapy is needed. Drugs inhibiting cyclooxygenase (cox) have recently been found to have chemopreventive and antitumor activity and may potentiate the effects of chemotherapy. The purpose of this study was to determine whether cisplatin combined with the cox-inhibitor piroxicam would induce remission more frequently than cisplatin alone in a relevant animal model of human invasive TCC. METHODS: Pet dogs with naturally occurring, histopathologically confirmed, measurable TCC of the urinary bladder were randomized to receive cisplatin (60 mg/m2 i.v. every 21 days) or cisplatin (same dosage) combined with piroxicam (0.3 mg/kg orally every 24 h). Complete staging was performed prior to and at 6-week intervals during therapy. RESULTS: After eight dogs had been evaluated in each treatment group, a significant difference in remission rate was noted (Fisher's Exact test, P < 0.004). Tumor responses in the cisplatin/piroxicam group included two complete remissions (CR), four partial remissions (PR), two stable disease (SD), and no progressive disease (PD). Tumor responses to cisplatin alone in eight dogs were no CR, no PR, four SD, and four PD. Six additional dogs were treated with cisplatin/piroxicam, and in total 10 of 14 dogs had remission (two CR, eight PR). Renal toxicity of cisplatin/ piroxicam was frequent and dose limiting. CONCLUSIONS: Cisplatin/piroxicam induced remission more frequently than cisplatin alone in a canine model of human invasive TCC. Strategies to reduce renal toxicity need to be developed prior to evaluation of cisplatin/piroxicam in humans or general use of this treatment in pet dogs. PMID- 11021741 TI - Effect of 5-(phenylselenenyl)acyclouridine, an inhibitor of uridine phosphorylase, on plasma concentration of uridine released from 2',3',5'-tri-O acetyluridine, a prodrug of uridine: relevance to uridine rescue in chemotherapy. AB - PURPOSE: The purpose of this investigation was to study the effects of combining oral 5-(phenylselenenyl)acyclouridine (PSAU) with 2',3',5'-tri-O-acetyluridine (TAU) on the levels of plasma uridine in mice. PSAU is a new lipophilic and potent inhibitor of uridine phosphorylase (UrdPase, EC 2.4.2.3), the enzyme responsible for uridine catabolism. PSAU has 100% oral bioavailability and is a powerful enhancer of the bioavailability of oral uridine. TAU is a prodrug of uridine and a far superior source of uridine than uridine itself. METHODS: Oral TAU was administered to mice alone or with PSAU. The plasma levels of uridine and its catabolites as well as PSAU were measured using HPLC and pharmacokinetic analysis was performed. RESULTS: Oral administration of 2000 mg/kg TAU increased plasma uridine by over 250-fold with an area under the curve (AUC) of 754 micromol x h/l. Coadministration of PSAU at 30 and 120 mg/kg with TAU further improved the bioavailability of plasma uridine resulting from the administration of TAU alone by 1.7- and 3.9-fold, respectively, and reduced the Cmax and AUC of plasma uracil. CONCLUSION: The exceptional effectiveness of PSAU plus TAU in elevating and sustaining a high plasma uridine concentration could be useful in the management of medical disorders that are remedied by administration of uridine, as well as the rescue or protection from host toxicities of various chemotherapeutic pyrimidine analogues. PMID- 11021742 TI - The role of caspase family protease, caspase-3 on cisplatin-induced apoptosis in cisplatin-resistant A431 cell line. AB - PURPOSE: Cisplatin (cis-diamminedichloro-platinum(II), CDDP) has been reported to induce apoptosis in cancer cells. the mechanism of the apoptosis in cancer cells induced by CDDP is still unclear. Recent studies have revealed that caspase family of cystine proteases play an important role in the regulation of several apoptotic processes. In this study, whether apoptosis induced by CDDP could be mediated by the activation of caspase-3, a caspase family protease, was investigated. METHODS: The CDDP-resistant subline A431/CDDP2 from the previously established human epidermoid carcinoma cell line A431 was used. The parent A431 cells (A431/P) and the A431/CDDP2 were exposed to CDDP with or without a caspase family protease inhibitor (Z-Asp-CH2-DCB), and cellular sensitivity to CDDP was determined. DNA fragmentation was then analyzed, and the caspase-3 protein levels determined by Western blotting following exposure of the cells to CDDP with or without Z-Asp-CH2-DCB. RESULTS: In the A431/P cells, the cytotoxicity of CDDP was clearly reduced by Z-Asp-CH2-DCB compared with its cytotoxicity in A431/CDDP2 cells. Furthermore, quantitative analysis of DNA fragmentation revealed that Z Asp-CH2-DCB inhibited DNA fragmentation induced by CDDP in A431/P cells, but not in A431/CDDP2 cells. Western blotting analysis demonstrated a marked reduction in procaspase-3 protein levels in A431/P cells treated with Z-Asp-CH2-DCB. In the A431/CDDP2 cells, procaspase-3 protein levels were no different with and without Z-Asp-CH2-DCB. CONCLUSIONS: These findings suggest that caspase-3 may mediate apoptosis induced by CDDP, and its induction could represent a novel approach to the effective treatment of malignant tumors. PMID- 11021743 TI - Preclinical oral antitumor activity of BMS-185660, a paclitaxel derivative. AB - PURPOSE: A water soluble paclitaxel derivative, BMS-185660, identified previously as having parenteral activity comparable with that of the parent drug, was evaluated for antitumor activity when given orally. METHODS: Staged subcutaneous (s.c.) tumor models of both murine and human origin were used for this purpose. RESULTS: BMS-185660 achieved levels of activity following oral administration which were comparable with those maximum effects obtained using intravenous (i.v.) paclitaxel. Consecutive daily oral administrations of BMS-185660 resulted in maximum gross log cell kill (LCK) values of 1.7-2.0 in two experiments involving the s.c. Madison 109 murine lung tumor model, which were comparable with the best effects of the derivative injected intravenously, and 0.3 to 0.9 LCK greater than the maximum effects obtained with i.v. paclitaxel; paclitaxel given orally was inactive. Against a human ovarian tumor model with developed resistance to cisplatin (A2780/ cDDP), oral BMS-185660 achieved a maximum LCK of 1.8 compared with i.v. paclitaxel, which produced a maximum 2.4 LCK. Also, in the human HCT-116 colon carcinoma model, oral BMS-185660 cured a maximum of seven of eight mice compared with six of seven mice cured with i.v. paclitaxel. The loss in potency between comparably effective intravenously and orally administered doses of BMS-185660 was about four- to five-fold, but since no drug-associated lethality was ever observed following the oral administration of the highest doses of BMS-185660, further dose escalation may have been tolerated. The intermediate metabolite between BMS-185660 and paclitaxel is BMS-181681. This compound was also evaluated orally and found not to be active versus s.c. M109, despite demonstrating good activity by the i.v. route. CONCLUSION: The comparable activities of both intravenously and orally administered BMS-185660 to intravenously administered paclitaxel, combined with the attribute of improved water solubility, provides a good basis for further derivative development. PMID- 11021745 TI - Characteristics and analysis of normal and leukemic stem cells: current concepts and future directions. AB - Acute myeloid leukemias (AML) are considered to be clonal disorders involving early hematopoietic progenitor cells. The recent advances in characterization of early stem cells give rise to the question whether it is possible to distinguish healthy progenitors from cells of the leukemic clone in leukemia patients. Differences and similarities in phenotype, genotype and biology are described for leukemic cells and normal hematological progenitors. Recent new insights into human stem cell development offer the perspective that distinction between benign and malignant progenitors might be possible in the future at a very early stage of maturation. PMID- 11021744 TI - A phase II pharmacodynamic study of pyrazoloacridine in patients with metastatic colorectal cancer. AB - PURPOSE: To perform a phase II trial of pyrazoloacridine (PZA), a novel DNA intercalator, in patients with metastatic colorectal carcinoma and no previous therapy. METHODS: PZA was administered at a dose of 750 mg/m2 intravenously over 3 h every 21 days. Pharmacokinetic studies to determine PZA plasma concentrations were performed. RESULTS: No responses were seen in 14 response-evaluable patients. Patients received a median of two cycles of PZA (range 1-6). Toxicity included neutropenia and neurologic side-effects, which were > or = grade III in 73% and 14%, respectively. High plasma concentrations of PZA (Cmax) correlated with low neutrophil counts (P = 0.04). CONCLUSIONS: PZA is inactive at this dose and schedule in colorectal cancer, and produces moderately severe toxicity. PMID- 11021746 TI - Origin and diversification of the clonogenic cell in multiple myeloma: lessons from the immunoglobulin repertoire. AB - The study of immunoglobulin genes in multiple myeloma over the last decade has provided important information regarding biology, ontogenetic assignment, disease evolution, pathogenic consequences and tumor-specific therapeutic intervention. Detailed analysis of VH genes has revealed the clonal relationship between switch variants expressed by the bone marrow plasma cell and myeloma progenitors in the marrow and peripheral blood. Regarding VH usage, a bias was found against the V4 34 gene encoding antibodies with cold agglutinin specificity (anti-I/i), thus explaining in part the absence of autoimmune phenomena in myeloma compared to other B cell lymphoproliferative disorders. However, in some studies a substantial number of cases analyzed were carrying the rearranged Humkappav325 Vkapppa gene, known to be over utilized by B cell chronic lymphocytic leukemia clones and possessing autoantibody binding activity. VH genes accumulate somatic hypermutations following a distribution compatible with antigen selection, but with no intraclonal heterogeneity. The analysis of Vkappa genes indicates a bias in usage of Vkappa family members; somatic hypermutation, in line with antigen selection, of the expressed Vkappa genes is higher than any other B cell lymphoid disorder. Similar conclusions were reached for Vlambda genes; in this case, the analysis raises the controversial issue of N nucleotide insertion at Vlambda Jlambda junctions, apparently as a result of TdT activity. A complementary imprint of antigen selection as evidenced by somatic hypermutation of either the VH or VL clonogenic genes has been observed. The absence of ongoing somatic mutations in either VH or VL genes gives rise to the notion that the cell of origin in myeloma is a post-germinal center memory B cell. PMID- 11021747 TI - Concurrent disruption of p16INK4a and the ARF-p53 pathway predicts poor prognosis in aggressive non-Hodgkin's lymphoma. AB - The INK4a/ARF locus at chromosome 9p21 encodes two structurally and functionally distinct molecules with tumor-suppressive properties. p16INK4a controls cell cycle progression by inhibiting phosphorylation of the retinoblastoma protein (Rb), while ARF prevents MDM2-mediated degradation of p53. By using a panel of PCR-based methods, we have examined the status of the p16INK4a, ARF and p53 genes in 123 cases of non-Hodgkin's lymphoma (NHL) at diagnosis. Alterations of one or more of these genes were detected in seven of 36 (19%) cases with low- to intermediate-grade histology, and in 35 of 87 (40%) cases with aggressive histology. For the aggressive lymphomas, the Kaplan-Meier estimate of overall survival for cases with disruption of either p16INK4a or the ARF-p53 pathway was not different from cases with retention of both pathways (5 year survival 45% vs 35%; P= 0.85), suggesting that selective inactivation of one of the pathways does not significantly influence overall survival. By contrast, the 5-year survival was only 7% for cases with concurrent disruption of p16INK4a and the ARF-p53 pathway vs 38% for cases with retention of one or both pathways (P = 0.005). Similar results were obtained when the analysis was confined to diffuse large B cell lymphomas (P= 0.019). On stepwise multivariate regression analysis including factors from the international prognostic index, concurrent disruption of p16INK4a and the ARF-p53 pathway was an independent negative prognostic factor in NHL with aggressive histology (P = 0.006). Our results suggest that the compound status of the p16INK4a and ARF-p53 pathways is a major determinant of outcome in NHL. PMID- 11021748 TI - Treatment of childhood acute myelogenous leukemia with an intensive regimen (AML 87) that individualizes etoposide and cytarabine dosages: short- and long-term effects. AB - The purpose of this study was to assess the feasibility and efficacy of a treatment regimen for pediatric acute myelogenous leukemia (AML) that uses four rotating drug pairs and adjusts dosages of etoposide and cytarabine to target specific plasma concentrations. Thirty-one girls and 27 boys (median age, 9.7 years) with de novo AML were treated on the protocol. Six cycles of chemotherapy were planned. Cycles 1 to 4 comprised the drug combinations cytarabine plus etoposide, cytarabine plus daunomycin, etoposide plus amsacrine, and etoposide plus azacitidine, respectively. For cycles 5 and 6, the first two combinations were repeated. Dosages were adjusted to achieve plasma concentrations of 1.0 microM +/- 0.1 microM cytarabine and 30 microM +/- 0.3 microM etoposide. Forty four patients (76%) entered complete remission. Of those, 24 have had relapses; 23 remain alive in first or subsequent remission. The 5-year event-free survival (EFS) estimate was 31.0% +/- 5.9%; the 5-year survival estimate was 41.4% +/- 6.3%. Six patients (10%) died of the toxic effects of therapy. Severe neutropenia occurred in all cycles. Long-term complications of therapy included hepatitis C, cardiac insufficiency, and hearing loss. Adjustment of cytarabine and etoposide dosage was feasible for achieving targeted plasma drug concentrations; however, the potential clinical efficacy of this approach was offset by substantial acute and long-term toxicity. PMID- 11021749 TI - Involvement of CD95-independent caspase 8 activation in arsenic trioxide-induced apoptosis. AB - Arsenic trioxide (As2O3)-treatment is effective in acute promyelocytic leukemia (APL) patients with t(15;17). Clinically achievable concentrations of As2O3 induce apoptosis in NB4, an APL cell line, in vitro. Here, to study the mechanism of As2O3-induced apoptosis, we established an As2O3-resistant subline, NB4/As. Growth of NB4/As was inhibited by 50% after 2 day-treatment (IC50) at 1.6 microM As2O3, whereas IC50 of NB4 was 0.3 microM. Degradation of PML-RARalpha and change of the PML-subcellular localization were similarly induced by As2O3 in NB4 and NB4/As, suggesting that their contribution to apoptosis is small. Treatment with 1 microM As2O3 induced the activation of caspase 3 as well as a loss of mitochondrial transmembrane potential (deltapsim) in NB4 but not in NB4/As. Caspase 8 and Bid were also activated by As2O3 in NB4 but not in NB4/As. In NB4, an inhibitor of caspase 8 blocked not only the activation of caspase 3 but also the loss of deltapsim. Neither cell line expressed CD95/Fas, and agonistic anti Fas antibody (CH-11) failed to cause apoptosis. Neither antagonistic anti CD95/Fas antibody nor anti-Fas ligand antibodies influenced the As2O3-induced apoptosis. NB4/As had a higher concentration of intracellular glutathione (GSH) than NB4 (96 vs 32 nmol/mg). Reduction of the GSH level by buthionine sulfoxide (BSO) completely restored the sensitivity to As2O3 in NB4/As. Furthermore, caspase activation and the loss of deltapsim were recovered by combination treatment with BSO. These findings suggest that the As2O3 treatment activates caspase 8 in a CD95-independent but GSH concentration-dependent manner. In combination with BSO, As2O3 might be applied to therapy of leukemia/cancers which are insensitive to the clinically achievable concentrations of As2O3. PMID- 11021750 TI - Colony-forming unit-megakaryocyte (CFR-meg) numbers and serum thrombopoietin concentrations in thrombocytopenic disorders: an inverse correlation in myelodysplastic syndromes. AB - We studied both serum-free colony-forming unit-megakaryocyte (CFU-meg) numbers and serum thrombopoietin (TPO) levels in 14 patients with aplastic anemia (AA), 37 patients with myelodysplastic syndromes (MDS) and 23 patients with idiopathic thrombocytopenic purpura (ITP) to assess thrombopoiesis in these thrombocytopenic disorders. The mean CFU-meg numbers were lower in AA and MDS patients (10.7 +/- 11.4 and 42.3 +/- 58.5/10(5) BMLD cells) than in healthy controls (103.1 +/- 57.3/10(5) BMLD cells) (P < 0.0001 and P= 0.0053, respectively), although they were distributed variably in MDS. ITP patients showed higher CFU-meg numbers (223.2 +/- 143.5/10(5) BMLD cells) (P= 0.017). The mean TPO concentrations were higher in both AA (986.8 +/- 500.8 pg/ml) and MDS patients (838.2 +/- 639.1 pg/ml) than in healthy controls (80.7 +/- 38.8 pg/ml) (P < 0.0001), although they were distributed from high to low in MDS. ITP patients showed a slight elevation of TPO (123.1 +/- 55.3 pg/ml) P = 0.0106). The TPO levels was inversely correlated to both platelet counts and CFU-meg numbers (correlative coefficient (CC): -0.719 and -0.682, P < 0.0001) in AA, but not in ITP. In MDS, the inverse correlation to TPO was stronger in CFU-meg (CC: -0.678, P < 0.0001) than in platelet counts (CC: -0.538, P = 0.0014), suggesting that CFU-meg plays an important role in regulating TPO production in this heterogenous disorder. CFU meg and TPO may provide useful information for understanding thrombopoiesis of MDS, especially for application of TPO. PMID- 11021751 TI - Elevated MLF1 expression correlates with malignant progression from myelodysplastic syndrome. AB - MLF1 is a novel protein identified as the NPM-MLF1 chimeric protein produced by a t(3;5)(q25.1;q34) chromosomal translocation, which is associated with myelodysplastic syndrome (MDS), often prior to acute myeloid leukemia (AML), except for M3. The clinical features of t(3;5)-positive myeloid disorders suggest that this chimeric protein is involved in dysregulation of progenitor cells with the capability to differentiate into multiple lineages. So far, involvement of wild-type MLF1 in hematopoiesis or in leukemogenesis has not been fully investigated. In the present study, 65 patients with AML and 44 patients with MDS were tested for the expression of MLF1 using the quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. A significantly higher level of MLF1 expression (ratio of MLF1/beta-actin mRNA >0.4) was readily detected in seven of 65 patients with de novo AML, three of 12 with post-MDS AML and seven of 44 with MDS, but not in any patients with ALL (n = 18). According to the FAB classification, high levels of MLF1 were found in patients with relatively immature subtypes of AML (M1, M2, M6 and M7) and high risk MDS (RAEB and RAEB-T). These findings indicate that the pattern of MLF1 expression is identical to the clinical morphology appearing in the t(3;5)-positive myeloid disorders and is correlated to the MDS-associated AML and transformation phase of MDS in t(3;5)-negative myeloid disorders. A CD34+ population of normal bone marrow cells preferentially expressed MLF1 with obviously decreasing levels of expression during maturation. Therefore, MLF1 normally functions in multi-potent progenitor cells and its dysregulation may take part in leukemogenesis from MDS. PMID- 11021752 TI - Constitutive activation of FLT3 stimulates multiple intracellular signal transducers and results in transformation. AB - Aberrant expression of FLT3 has been found in most cases of B-lineage ALL and AML, and subsets of T cell ALL, CML in blast crisis and CLL. In 20% of patients with AML the receptor has small internal tandem duplications of the juxtamembrane region which appear to contitutively activate the receptor. To investigate whether FLT3 activation could play a role in leukemia, we generated a constitutively activated FLT3 by fusing its cytoplasmic domain to the helix-loop helix domain of TEL in analogy to the fusion that occurs with TEL-PDGFR in CMML. In vitro translation assays demonstrated oligomerization and intrinsic tyrosine kinase activity of the TEL-FLT3 chimeric receptor. Constitutively activated TEL FLT3 conferred IL-3 independence and long-term proliferation to transfected Ba/F3 cells. Immunoblot analyses showed that JAK 2, STAT 3, STAT 5a, STAT 5b and CBL were tyrosine-phosphorylated in TEL-FLT3 expressing Ba/F3 cells in the absence of IL-3. These data suggest a possible role for the JAK/STAT pathway in FLT3 signaling. Transplantation of TEL-FLT3 expressing Ba/F3 cells into syngeneic mice caused mortality in all mice by 3 weeks after injection. Histopathologic analysis demonstrated a massive infiltration of mononuclear cells in the liver, spleen and bone marrow. The mimicking of naturally occurring TEL fusions provides an approach to assess aspects of the biology of activated FLT3, or other receptor type tyrosine kinases (RTKs) in leukemic transformation. PMID- 11021753 TI - The interleukin-3 receptor alpha chain is a unique marker for human acute myelogenous leukemia stem cells. AB - Recent studies suggest that the population of malignant cells found in human acute myelogenous leukemia (AML) arises from a rare population of leukemic stem cells (LSCs). LSCs have been documented for nearly all AML subtypes and have been phenotypically described as CD34+/CD38- or CD34+/HLA-DR-. Given the potentially critical role of these primitive cells in perpetuating leukemic disease, we sought to further investigate their molecular and cellular characteristics. Flow cytometric studies using primary AML tissue showed that the interleukin-3 receptor alpha chain (IL-3Ralpha or CD123) was strongly expressed in CD34+/CD38- cells (98 +/- 2% positive) from 16 of 18 primary specimens. Conversely, normal bone marrow derived CD34+/CD38- cells showed virtually no detectable expression of the CD123 antigen. To assess the functional role of IL-3Ralpha positive cells, purified CD34+/CD123+ leukemia cells were transplanted into immune deficient NOD/SCID mice. These experiments showed that CD123+ cells were competent to establish and maintain leukemic populations in vivo. To begin to elucidate a biological role for CD123 in leukemia, primary AML samples were analyzed with respect to signal transduction activity in the MAPK, Akt, and Stat5 pathways. Phosphorylation was not detected in response to IL-3 stimulation, thereby suggesting CD123 is not active in conventional IL-3-mediated signaling. Collectively, these data indicate that CD123 represents a unique marker for primitive leukemic stem cells. Given the strong expression of this receptor on LSCs, we propose that targeting of CD123 may be a promising strategy for the preferential ablation of AML cells. PMID- 11021754 TI - Differentiation commitment and regulator-specific granulocyte-macrophage maturation in a novel pro-B murine leukemic cell line. AB - The cloned pro-B-lymphocyte murine leukemic cell line GB2, was established from a leukemic Max41 x Emu-myc double transgenic mouse. Its Igh alleles are rearranged and its surface markers are primarily B-lymphoid, but a small proportion of the cells also express surface Gr-1 and some cells develop the morphology of maturing granulocytes. The cell line grows continuously in suspension culture without the addition of growth factors, but expresses mRNA for M-CSF, TPO and Flt-3-ligand. When stimulated in agar cultures by GM-CSF, G-CSF, M-CSF, IL-3, SCF, IL-6, leukemia inhibitory factor (LIF), IL-5 or IFNgamma, GB2 cells generated blast colonies or colonies of maturing granulocytes and macrophages. There was a striking similarity in colony types, relative colony numbers and maturation of colony cells to those formed by normal bone marrow cells in response to the same stimuli. GB2 blast colony-forming cells exhibited self-renewal as well as an ability to form granulocyte-macrophage colony-forming progeny, with evidence that a hierarchical sequence of clonogenic cells is generated in the cell line even after subcloning. Factor-specific maturation was clearly initiated by the action of the added growth factors. In contrast, FACS-sorting experiments showed that commitment to various types of colony-forming cell occurs in maintenance suspension cultures in the apparent absence of potentially relevant growth factors. PMID- 11021755 TI - The effects of dose, route of administration, drug scheduling and MDR-1 gene transfer on the genotoxicity of etoposide in bone marrow. AB - We have used the bone marrow micronucleus assay (BMMN) as a measure of clastogenicity, in response to etoposide exposure in murine bone marrow. Oral delivery of etoposide resulted in a reduced number of micronucleated polychromatic erythrocytes (MPE) relative to the same dose delivered intraperitoneally (P < 0.001). Daily fractionation of the oral schedule of etoposide led to a more than six-fold increase in cumulative MPE frequency over that observed with the same total, unfractionated dose, with the potency of the response increasing with serial exposure (r = 0.79). Retrovirally-mediated expression of MDR1 in murine bone marrow resulted in partial protection against the clastogenic activity of etoposide relative to mock transduced control mice. The model system developed has indicated a variety of factors able to influence the genotoxicity of etoposide. It should now be possible to further exploit this model in order to define other factors governing haemopoietic sensitivity to etoposide. PMID- 11021756 TI - Karyotypic dissection of Hodgkin's disease cell lines reveals ectopic subtelomeres and ribosomal DNA at sites of multiple jumping translocations and genomic amplification. AB - Although the neoplastic significance of the chromosome changes widespread in Hodgkin's disease (HD) remains obscure, a distinct cytogenetic picture has emerged combining aneuploidy with structural rearrangements clustered at certain breakpoints. Notably absent are the recurrent chromosome translocations which distinguish other hematopoietic neoplasms and serve as clues to underlying oncogene alterations. The paucity of neoplastic cells in HD biopsies hinders detailed chromosome analysis. As an alternative, we investigated a panel of well characterized cell lines by classical and molecular cytogenetics, using single gene and subtelomeric probes, including three autologous HD examples (HDLM-1/2/3) analyzed by 'spectral karyotyping' - the first complete HD karyotype to be documented. Although complex, most rearrangements in HDLM cells arose in vivo and included few rare but many typical HD breakpoints, notably at the r(ibosomal)DNA regions. Two types of genomic rearrangement involving DNA repeats were conspicuous: insertion and genomic amplification/coamplification of rDNA-the first genomic rDNA rearrangements to be reported in a tumor cell, and the first example of multiple 'jumping translocations' (JT). Of four subtelomeric microsatellite repeats tested in HDLM cells, three exhibited interstitial sites at JT, of which two (at 5qter and 9pter) were respectively associated with deletion of the 5q31-32 myeloid region, and coamplification of a recently described HD-recurrent amplicon at 9p2 together with transcriptionally silent rDNA. Altogether, three out of four HD cell lines carried interstitial 9p subtelomeres and rDNA rearrangements. Taken together, these data suggest tumorigenic rearrangements may be facilitated by 'hitchhiking' along with mobile DNA repeat sequences which may target gene rearrangement at 9p in HD. Southern analysis of parallel rearrangements within rDNA intergenic spacers in HDLM cells highlighted several at, or near, retroposons. As well as validating HD cell lines as cytogenetic models, and resources for identifying genes rearranged in HD, our findings warrant further investigation of the roles of DNA repeat sequences, notably subtelomeric microsatellites, rDNA spacer sequences and retroposons as facilitators and markers of tumor-gene rearrangement. PMID- 11021757 TI - Lack of benefit of CD34+ cell selected over non-selected peripheral blood stem cell transplantation in multiple myeloma: results of a single center study. AB - In order to determine the clinical impact of CD34+ cell selected autologous transplantation in multiple myeloma (MM), we have performed a retrospective case controlled analysis comparing 21 MM patients receiving high-dose melphalan and autologous transplantation with CD34+ peripheral blood stem cells (PBSC) as front line therapy to 21 control patients receiving unselected products. Case matching was performed using the following criteria: age and beta2-microglobulin at diagnosis and disease status at the time of transplantation. Both cohorts were homogeneous in term of induction treatment and conditioning regimen. Patients were collected for CD34+ selection after priming with G-CSF alone. Significantly fewer CD34+ cells/kg were infused to patients in the selected group as compared to patients in the control group: 2.2 (range 0.5-14.3) vs 9.4 (range 1.1-15) (P < 0.001). The median time to neutrophil recovery > or =0.05 x 10(9)/l was 10 days for the CD34+ group and 9.5 days for the control group (P = 0.357). The median time to platelet recovery > or = 20 x 10(9)/l was 9 days for the CD34+ group and 4.5 days for the control group (P = 0.005). Response rates were comparable in both groups (85.7% in the CD34+ group vs 90.4% in the control group). At 3 years, event-free survival (32% in the CD34+ group vs 39% in the control group) and overall survival (85% in the CD34+ group vs 79% in the control group) were not significantly different. Finally, use of unselected products dramatically reduced the cost of the transplantation procedure. This study shows that CD34+ cell selected autologous transplantation is more expensive than transplantation with unselected products and does not improve the clinical outcome of patients with MM. PMID- 11021758 TI - Biological significance of the expression of HIV-related chemokine coreceptors (CCR5 and CXCR4) and their ligands by human hematopoietic cell lines. AB - The aim of this study was to learn more about the role of the HIV-related chemokine-chemokine receptor axes in human hematopoiesis. To address this issue we phenotyped 35 selected hematopoietic cell lines for the expression of CD4, CXCR4 and CCR5. We next evaluated the functionality of these chemokine receptors by calcium flux and chemotaxis assays, and by the ability of SDF-1, MIP-1alpha, MIP-1beta and RANTES to influence the growth of the cells expressing CXCR4 and/or CCR5. Lastly, we examined whether human hematopoietic cell lines may secrete some HIV-related chemokines, and whether endogenously secreted chemokines might interfere with the infectability. of hematopoietic cells by X4 and R5 HIV strains. These results demonstrate that: (1) HIV-related receptors are widely expressed on human hematopoietic cell lines; (2) stimulation of CXCR4 by SDF-1 induces calcium flux and chemotaxis in several hematopoietic cell lines more efficiently than stimulation of CCR5 by receptor-specific beta-chemokines; (3) chemokines do not regulate proliferation of the hematopoietic cells; and finally (4) infectability of the hematopoietic cells by HIV-1 may be auto-modulated by endogenously secreted chemokines. These data shed more light on the role of HIV related chemokine-chemokine receptors axes in human hematopoiesis and interaction of hematopoietic cells with HIV. PMID- 11021759 TI - Positive and negative regulation of apoptotic pathways by cytotoxic agents in hematological malignancies. AB - Most chemotherapeutic drugs can induce tumor cell death by apoptosis. Analysis of the molecular mechanisms that regulate apoptosis has indicated that anticancer agents simultaneously activate several pathways that either positively or negatively regulate the death process. The main pathway from specific damage induced by the drug to apoptosis involves activation of caspases in the cytosol by pro-apoptotic molecules such as cytochrome c released from the mitochondrial intermembrane space. At least in some cell types, anticancer drugs also upregulate the expression of death receptors and sensitize tumor cells to their cognate ligands. The Fas-mediated pathway could contribute to the early steps of drug-induced apoptosis while sensitization to the cytokine TRAIL could be used to amplify the response to cytotoxic drugs. The Bcl-2 family of proteins, that includes anti- and pro-apoptotic molecules, regulates cell sensitivity mainly at the mitochondrial level. Anticancer drugs modulate their expression (eg through p53-dependent gene transcription), their activity (eg by phosphorylating Bcl-2) and their subcellular localization (eg by inducing the translocation of specific BH3-only pro-apoptotic proteins). Very early after interacting with tumor cells, anticancer drugs also activate lipid-dependent signaling pathways that either increase or decrease cell ability to die by apoptosis. In addition, cytotoxic agents can activate protective pathways that involve activation of NFkappaB transcription factor, accumulation of heat shock proteins such as Hsp27 and activation of proteins involved in cell cycle regulation. This review discusses how modulation of the balance between noxious and protective signals that regulate drug-induced apoptosis could be used to improve the efficacy of current therapeutic regimens in hematological malignancies. PMID- 11021760 TI - Quality assurance in RT-PCR-based BCR/ABL diagnostics--results of an interlaboratory test and a standardization approach. AB - Here we describe the results of an interlaboratory test for RT-PCR-based BCR/ABL analysis. The test was organized in two parts. The number of participating laboratories in the first and second part was 27 and 20, respectively. In the first part samples containing various concentrations of plasmids with the ela2, b2a2 or b3a2 BCR/ABL transcripts were analyzed by PCR. In the second part of the test, cell samples containing various concentrations of BCR/ABL-positive cells were analyzed by RT-PCR. Overall PCR sensitivity was sufficient in approximately 90% of the tests, but a significant number of false positive results were obtained. There were significant differences in sensitivity in the cell-based analysis between the various participants. The results are discussed, and proposals are made regarding the choice of primers, controls, conditions for RNA extraction and reverse transcription. PMID- 11021761 TI - Breakpoint analysis by fluorescence in situ hybridization in myelodysplastic syndromes with t(3;12)(q26;p13) and expression of EVI1. PMID- 11021762 TI - A sensitive, non-radioactive and fast method for detection of JNK/SAPK activity in leukemic T cells. PMID- 11021763 TI - Preliminary evidence that Ghrelin, the natural GH secretagogue (GHS)-receptor ligand, strongly stimulates GH secretion in humans. AB - An endogenous ligand for the GH secretagogue-receptor (GHS-R) has been recently purified from rat and human stomach and named Ghrelin. It has been demonstrated that Ghrelin specifically stimulates GH secretion from rat pituitary cells in culture as well as in rats in vivo. In this preliminary study, in 4 normal adults [age (mean+/-SE): 28.6+/-3.5 yr; body mass index (BMI): 22.3+/-2.1 kg/m2] we administered 1.0 microg/kg Ghrelin or GHRH-29 to compare their GH-releasing activities in humans. In all subjects Ghrelin induced a prompt, marked and long lasting increase in circulating GH levels (peak: 107.9+/-26.1 microg/l; AUC: 6503.1+/-1632.7 microg/l/h). The GH response to Ghrelin was clearly higher (p<0.05) than that after GHRH (peak: 22.3+/-4.5 microg/l; AUC: 1517.5+/-338.4 microg/l/h). In conclusion, this preliminary study shows that Ghrelin exerts a strong stimulatory effect on GH secretion in humans releasing more GH than GHRH. PMID- 11021764 TI - Post-partum thyroiditis in South Brazil presenting as thyrotoxicosis: prevalence and risk factors. AB - The prevalence of post-partum thyroiditis (PPT) has been reported in several countries (1.9 to 16.7%) but is not known in Brazil. Several factors have been associated to its development, such as a female sex of the newborn, PPT in a previous pregnancy, a family history of thyroid disease and cigarette smoking. To investigate the prevalence of PPT and its risk factors in a southern Brazilian city, a three-cross-sectional observation study was performed. PPT was diagnosed in 14/284 subjects (5.3%) and all cases had thyrotoxicosis (13 sub-clinical and one clinical). Serum total T4 and free T4 were higher and serum TSH was lower in PPT subjects. Anti-thyroid antibodies were positive in 16.7% of PPT subjects and in 4.5% of those with no thyroid dysfunction. Goiter was identified in 14.3% of PPT subjects and in 15% of no PPT subjects. Thyroid was hardened more frequently in PPT subjects (21.4%) than in others (5.2%). Male sex of the newborn was associated to PPT, increasing 11 times the risk of PPT. Cigarette smoking was associated to PPT in group II subjects. There was no clinical sign or symptom able to contribute to this diagnosis, except the presence of hardened thyroid. Based on these findings, PPT, manifesting itself as mild thyrotoxicosis, is a common problem in southern Brazil and is associated to male sex of the newborn. PMID- 11021765 TI - Epidermal growth factor influenced by opioid peptides in immature rat uterus. AB - The aim of the present experiment was to investigate the effect of [D-Met2,Pro5] enkephalinamide (ENK) implantation on the development of the uterus during 8-33 days of age and the involvement of epidermal growth factor (EGF) in the effect. Administration of ENK was attained by osmotic minipumps (5 microg/h) implanted intraperitoneally. ENK resulted in a decrease in the EGF content of the uterus, which was already significant after 48 h of the implantation. The DNA content 24 and 48 h after the treatment decreased, no change at 72 h was found, however the protein/DNA ratio on the effect of ENK treatment was significantly decreased at this time in all examined age groups. High affinity and lower capacity competitive naloxone binding sites were demonstrated in the membrane fraction of the uteri. Seventy-two h after ENK treatment the binding capacity of these sites significantly dropped. The present results suggest a novel multiple interaction between estrogen and two probably paracrine hormones, EGF and opioid peptide, in the regulation of growth and development of the uterus. PMID- 11021766 TI - Circulating interleukin-6 significantly correlates to thyroid hormone in acute myocardial infarction but not in chronic heart failure. AB - To investigate relationships between thyroid states and the cardiac endocrine system, we analyzed thyrotropin (TSH), thyroid hormone, plasma levels of interleukin-6 (IL-6) and brain natriuretic peptide (BNP) in 50 patients with chronic heart failure (CHF), in 30 patients with heart failure from acute myocardial infarction (AMI), and in 15 controls. Plasma levels of IL-6 and BNP in both CHF and AMI were significantly elevated, while free triiodothyronine (FT3) was significantly decreased compared to controls. FT3/free thyroxine (FT4) ratio was significantly decreased in CHF but not in AMI compared to controls. In CHF, diuretic treatment diminished circulating BNP but not IL-6, while diuretic treatment increased FT3/FT4 ratio. In AMI, FT3/FT4 ratio was significantly decreased 72 h compared to 12 h after the onset of AMI, while BNP and IL-6 were significantly increased 72 h compared to 12 h after the onset of AMI. In both CHF and AMI, BNP significantly correlated with FT4. On the other hand, significant correlations between IL-6 and FT3, and between IL-6 and FT3/FT4 ratio were detected in AMI but not in CHF. This preliminary study suggests that IL-6, BNP and thyroid hormone reflect ventricular dysfunction in both acute and chronic heart failure, and that IL-6 significantly relates to circulating thyroid hormone in AMI but not in CHF. PMID- 11021767 TI - Prevalence of the sleep apnea syndrome in acromegaly population. AB - The prevalence of sleep apnea syndrome (SAS) in acromegaly is high. Consequences of SAS are serious and are associated with increased morbidity and mortality. The aim of this study was to assess the relative frequency and predictive factors for SAS in a group of patients with acromegaly (n=55). The presence of SAS was evaluated using the Polymesam device. Hormonal and clinical examination consisted of assessment of growth hormone, insulin-like growth factor I plasma levels, body mass index (BMI), neck circumference, age, sex, treatment modes of acromegaly and ear, nose and throat (ENT) examination. The relative frequency of SAS in our group of patients with acromegaly was 75%. Independent predictors of SAS were: increased activity of acromegaly, higher age and neck circumference. No association between SAS and BMI and ENT findings was observed. The role of gender was controversial. PMID- 11021768 TI - Normal IGF-I and enhanced IGFBP-3 response to very low rhGH dose in patients with dilated cardiomyopathy. AB - Well-nourished patients with dilated cardiomyopathy (DCM) show slight reduction of mean basal IGF-I levels which, however, display a response to a rhGH dose as low as 5.0 microg/kg/day similar to that of age-matched control subjects (CS). To further investigate peripheral GH sensitivity, we studied the IGF-I and IGFBP-3 responses to 4-day s.c. 2.5 microg/kg/day rhGH administration, the lowest effective dose able to increase IGF-I levels in normal subjects, in 10 DCM patients [age (mean+/-SE): 57.6+/-1.0 yr, body mass index (BMI): 24.0+/-1.2 kg/m2, left ventricular ejection fraction: 26.2+/-3.2%, NYHA (New York Heart Association): I/0, II/4, III/4, IV/2] and in 9 age-matched healthy CS (age: 55.3+/-1.2 yr, BMI: 23.7+/-1.8 kg/m2). Basal IGF-I levels in DCM were lower though not significantly than those in CS (147.7+/-9.8 vs 174.7+/-17.0 microg/l). Basal IGFBP-3 levels in DCM were similar to those in CS (3.1+/-0.3 vs 2.7+/-0.2 mg/l). In CS 4-day rhGH increased IGF-I levels (222.4+/-14.9 microg/l; p<0.01 vs baseline) but did not modify IGFBP-3 levels (3.0+/-0.2 mg/l). In DCM IGF-I levels were increased by 4-day rhGH administration (175.7+/-11.0 microg/l; p<0.05 vs baseline) with a similar percent extent than in CS. On the other hand, in DCM, but not in CS, 4-day rhGH significantly increased IGFBP-3 levels (3.5+/-0.3 mg/l; p<0.05 vs baseline). Therefore, in conclusion, testing with the lowest effective rhGH dose further suggest that peripheral GH sensitivity in well-nourished DCM is preserved. On the other hand, DCM patients show enhanced IGFBP-3 sensitivity to stimulation by rhGH. PMID- 11021769 TI - Pivagabine decreases stress-related hormone secretion in women with hypothalamic amenorrhea. AB - Stress-induced neuroendocrine activities influence the regulation of endocrine glands and axes. Weight loss-related hypothalamic amenorrhea is a typical stress induced physiopathological condition. It is characterized by increased adrenal cortex activation and by reduced GH, LH, FSH and gonadal steroid hormone levels. The aim of the present study was to investigate the effects of pivagabine, a neurotropic drug (1800 mg/day for 7 days) or placebo administration on ACTH, cortisol, GH, LH, FSH and PRL plasma levels in patients with hypothalamic amenorrhea related to weight loss. Hormonal parameters and the pulsatile release of cortisol (6-hour pulsatility, sampling every 10 minutes) were evaluated before and after 7 days of treatment. Pivagabine administration significantly reduced mean plasma ACTH (from 21.7+/-1.7 to 15.4+/-1.2 pg/ml, p<0.05) and cortisol levels (from 12.2+/-0.7 to 9.7+/-0.7 ng/ml, p<0.05) and increased GH levels (from 1.4+/-0.5 to 3.0+/-0.9 ng/ml, p<0.05). A significant reduction of cortisol pulse amplitude was observed (p<0.01) while no change in pulse frequency occurred. No changes were observed in placebo-treated subjects. LH, FSH and PRL levels were not modified by placebo or pivagabine administration. In conclusion, in patients with hypothalamic amenorrhea related to weight loss pivagabine induced a significant decrease of cortisol secretion and an increase of GH release by pivagabine administration, suggesting that this drug exerts a specific neuroendocrine modulatory role. PMID- 11021770 TI - Case of an ivory vertebra. AB - The differential diagnosis of an osteoblastic vertebral lesion (ivory vertebra) includes metastatic prostate cancer, lung cancer, lymphoma, osteosarcoma and Paget's disease. We report a case of a man who was initially diagnosed with Paget's disease on vertebral biopsy. He failed to respond to conventional bisphosphate therapy. The review of the original biopsy specimen showed metastatic carcinoid tumor involving the bone marrow. The various features of carcinoid tumors metastasizing to the skeleton are briefly reviewed. PMID- 11021771 TI - Cushing's syndrome caused by ectopic corticotropin secretion by multiple peripheral pulmonary carcinoids and tumorlets of carcinoid type. AB - Multiple peripheral pulmonary carcinoid tumors or their smaller counterparts (tumorlets of carcinoid type) are the most unusual form of carcinoids as a cause of ectopic corticotropin syndrome. Only three case reports were found in the literature. We describe a 35 year-old female patient with ectopic corticotropin secretion due to multiple peripheral pulmonary carcinoid tumors and tumorlets. A high-dose dexamethasone suppression test result led to the diagnosis of Cushing's disease in our case. But no tumor was identified on sella imaging and bilateral inferior petrosal sinus sampling was non-diagnostic. Computed tomography of the lungs revealed multiple acinar-nodular parenchymal infiltrations confined to the left lung. Corticotropin-dependent hypercortisolism persisted after bilateral adrenalectomy. A second operation was necessary to remove the hyperplastic adrenal remnants. Meanwhile, computed tomography findings of the thorax were unchanged. We decided to explore these nodules by open lung biopsy. During the procedure multiple nodules ranging 12 to 3 mm in diameter scattered throughout the left lung were observed and left pneumonectomy was performed. Histopathological diagnosis was multiple peripheral carcinoid tumors and tumorlets of carcinoid type showing positive immunostaining with corticotropin. This observation emphasizes a rare form of carcinoids as a cause of ectopic corticotropin secretion and its unusual response to high dose dexamethasone suppression test. PMID- 11021772 TI - Venous thrombo-embolism following inferior petrosal sinus sampling in Cushing's disease. AB - Bilateral inferior petrosal sinus sampling for ACTH with corticotrophin releasing hormone stimulation has become an established test in differentiating pituitary Cushing's disease from Cushing's syndrome due to ectopic ACTH secretion. We report two patients with Cushing's disease who developed thromboembolic complications soon after inferior petrosal sinus sampling. We discuss the possible mechanisms leading to this complication in a syndrome in which thromboembolic complications are well recognized and highlight the need for consideration of prophylactic anticoagulation. PMID- 11021774 TI - Introduction: the advent of minimalism in neurosurgery. PMID- 11021773 TI - The PTH/PTHrP receptor in Jansen's metaphyseal chondrodysplasia. AB - JMC is a rare autosomal dominant form of short limb dwarfism characterized by asymptomatic hypercalcemia and skeletal deformities, despite low PTH and PTHrP levels. This rare disorder is likely to be caused by activating mutations in the PTH/PTHrP receptor leading to ligand-independent cAMP accumulation. The analysis of genetically altered mice which lack either PTHrP or the PTH/PTHrP receptor, as well as of transgenic mice in which the mutant receptor is targeted to the growth plate, has provided a molecular explanation for the severe skeletal abnormalities seen in JMC. In addition, the study of this rare human disorder has further elucidated the fundamental role played by the PTH/PTHrP receptor in mediating both the paracrine/autocrine actions of PTHrP in growth plate development and bone elongation, as well as the endocrine actions of PTH. The insight gained from the study of this human disease model is likely to continue to provide an important tool to define the cellular and molecular mechanisms that mediate the biological roles of the PTH, PTHrP and their receptor. PMID- 11021776 TI - Endoscopic carpal tunnel release: report of 146 cases. AB - OBJECTIVE: The purpose of this study is to describe one type of surgical treatment of carpal tunnel syndrome, namely endoscopic carpal tunnel release. MATERIALS AND METHODS: One hundred forty-six (146) cases of median nerve compression underwent the same-day procedure. RESULTS: Eighty-six percent (86%) were favorable, and 14% unfavorable. There was no damage caused to nerves, vascular structures, or tendons. CONCLUSIONS: Although overall long-term results of endoscopic carpal tunnel release appear similar to those of open release, patients who undergo endoscopic carpal tunnel release demonstrate a quicker functional recovery, less postoperative pain, and less scarring than open carpal tunnel release patients. PMID- 11021775 TI - Lumboperitoneal shunts. AB - BACKGROUND: To assess lumboperitoneal shunting as a simple method of relieving communicating hydrocephalus, treating pseudotumor cerebri, and resolving cerebrospinal fluid fistulae. METHOD: Three neurosurgeons at different hospitals report their combined experience over a 20-year period. RESULTS: Ease of insertion of the device and low rate of complications were reported in a combined series of 107 patients. CONCLUSIONS: Lumboperitoneal shunting is a technically simple and highly effective surgical technique for diversion of cerebrospinal fluid. PMID- 11021777 TI - Percutaneous microdecompressive endoscopic cervical discectomy with laser thermodiskoplasty. AB - OBJECTIVE: To study the surgical outcome of outpatient percutaneous microdecompressive endoscopic cervical discectomy with lower energy laser for shrinkage of disc material (thermodiskoplasty). METHOD: Since 1994, 200 patients with herniated cervical discs have presented at the authors' clinic, with unilateral radicular pain. The diagnosis was confirmed by MRI or CT, and EMG. RESULTS: At an average follow-up of 25 months, 94.5% of the cases had good-to excellent results. Eleven patients (5.5%) remained symptomatic, with persistent neck and upper extremity pain associated with paresthesias. There were no significant postoperative complications. Average time before returning to work was 10 days. CONCLUSIONS: Percutaneous microdecompressive endoscopic cervical discectomy with laser thermodiskoplasty has proven to be a safe and efficacious minimally invasive procedure. PMID- 11021778 TI - Arthroscopic microdiscectomy: an alternative to open disc surgery. AB - OBJECTIVE: To assess minimally invasive spinal surgery under endoscopic magnification and illumination (arthroscopic microdiscectomy) as a reliable alternative to open microsurgery for most herniated lumbar discs. METHOD: A total of 600 cases are evaluated retrospectively in terms of patient selection and technique. One series of 300 operations was performed by a key academician in the development of arthroscopic microdiscectomy. A second series of 300 patients was treated by a neurosurgeon in private suburban practice. RESULTS: In terms of patients self-evaluation, satisfactory outcome rates of 85-92% were realized. The patients considered brief intravenous anesthesia and same-day scheduling preferable to general anesthesia and hospitalization needed for open laminotomy and discectomy. Fewer than 2% of the cases required a second surgery. CONCLUSION: The authors are of the opinion that advantages include: (1) one-hour operative time, (2) negligible blood loss, (3) avoidance of significant scarring in the spinal canal, and (4) anterolateral fenestration of the annulus for continuing relief of intradiscal pressure and nerve root decompression. PMID- 11021779 TI - Percutaneous stereotactic radiofrequency thermal rhizotomy for the treatment of trigeminal neuralgia. AB - BACKGROUND: Trigeminal neuralgia (TN) is the most common cephalic neuralgia in people over the age of fifty, with a mean incidence of 4 per 100,000. Percutaneous stereotactic differential radiofrequency thermal rhizotomy (RTR) is a well-recognized surgical treatment for TN. The purpose of this study was to evaluate a management algorithm for TN and to evaluate the effectiveness of RTR for TN after failure of pharmacologic management. METHODS: Two hundred and fifty eight (258) patients underwent RTR from 1992-1996 and were prospectively evaluated. These patients were characterized by age, sex, side of the face and division(s) involved. Patients were evaluated for pain relief, recurrence requiring or not requiring re-operation, and the type and rate of complications. They were followed by serial clinical evaluation and telephone interview. Patients were grouped according to results: (A) Successful Result--excellent or good pain relief; (B) Unsuccessful Result--fair, poor or no pain relief. The RTR group was compared to historical controls. Follow-up ranged from 12-80 months (mean = 38 months). RESULTS: At early follow-up (defined as median postoperative period up to 6 months), pain relief that was excellent or good (successful results) occurred in 224/258 (87%). At long-term follow-up (> 6 months) recurrence of tic pain required re-operation in 31 patients (12%). In 37 patients (14%), recurrence of tic pain did not require re-operation. Dysesthesia developed in 20 patients (8%); corneal analgesia developed in 8 patients (3%). "Anesthesia dolorosa" developed in 5 patients (2%) and was medically managed. At the conclusion of the long-term follow-up period, 214/258 patients (83%) had excellent to good pain relief (successful result). There were no mortalities, no significant morbidity and a low rate of minor complications. CONCLUSION: With the use of this specific diagnostic and management algorithm, patients with TN can be successfully managed with RTR. PMID- 11021780 TI - Frameless stereotaxy of the brain. AB - Today's neurosurgical journals are replete with advertisements for systems designed to provide image guidance during surgery. These so-called "frameless" stereotactic systems provide the surgeon with navigational information, relating the location of instruments in the operative field to preoperative imaging data. Such information minimizes invasiveness by more accurately selecting the best trajectory to the lesion, ensures more precise identification of normal structures, and guides complete removal of a lesion. To achieve these goals, all of these systems utilize the stereotactic principle of co-registration of the patient with an imaging study. This review will trace the development of image guided surgery from its origins in frame-based stereotaxy to its current use as a surgical navigation methodology. A review of the more prevalent techniques and available systems will be presented, along with examples of specific applications of surgical navigation. Finally, some of the future directions of frameless stereotaxy will be discussed. PMID- 11021781 TI - Continuing experience with chemonucleolysis. AB - Chemonucleolysis using chymopapain is the least invasive technique used to treat a herniated lumbar pulposus. After 37 years of clinical experience, multiple clinical trials, a national multicenter, double-blind study mandated by the Food and Drug Administration, and heated controversy in the scientific community, the injection of chymopapain to treat herniated discs has (in appropriately selected patients) proven as successful as laminectomy, with fewer complications and the advantage of considerable cost savings. PMID- 11021782 TI - Anterior cervical discectomy without fusion or instrumentation: 25 years' experience. AB - OBJECTIVE: To assess a minimalist approach to anterior cervical discectomy without instrumentation or bone grafting. METHOD: A total of 530 operations performed during a 25-year period were evaluated in terms of technique and patient outcome. RESULTS: Advantages included minimal blood loss, little tissue swelling, less narcotic requirement, fewer days in the hospital, and earlier return to customary activity. During the 2-year follow-up period, a repeat surgical procedure was required in only 2% of the group. CONCLUSIONS: Eliminating spinal fusion from anterior cervical discectomy shortened the operation, reduced the number of surgical instruments needed, and eliminated complications associated with obtaining a graft from the iliac crest or maintaining the bone in situ. PMID- 11021783 TI - CT-guided needle procedures for brain lesions: 20 years' experience. AB - OBJECTIVE: To assess free-hand needle procedures with CT guidance for biopsy and drainage of brain lesions. METHOD: A series of 184 operations was carried out on 120 patients over a 20-year period. Sixty (60) cases of suspected neoplasm were biopsied; 40 cases of intracerebral hemorrhage and 20 cases of intracranial suppuration underwent drainage. RESULTS: The morbidity and mortality of the methods described were quite low due to steroid therapy, prophylactic antibiotics, and careful preparation of the access site. There was one complication of postoperative hemorrhage in a patient with metastatic melanoma. CONCLUSIONS: The simple and accurate techniques presented continue to have widespread application in the practice of neurosurgery and compare quite favorably with techniques using more elaborate equipment, stereotactic frames, and intraoperative monitoring. PMID- 11021785 TI - The evolution of percutaneous spinal endoscopy and discectomy: state of the art. AB - OBJECTIVE: The author relates his 7 years' experience with endoscopic spine surgery for lumbar disc herniations and conditions previously treated only with more invasive methods. MATERIALS AND METHODS: Five hundred (500) patients were treated with the Yeung endoscopic spine system, which features an endoscope with a 2.8 mm operating channel. The protocol included preoperative or intraoperative discography in all cases. Adjuvant therapies were employed in various clinical conditions when dictated by the visualized spinal pathology--KTP laser (Laserscope, San Jose, CA) in 100 patients, radiofrequency by electrothermal probe in 400 patients, chymopapain in 50 cases, and intraoperative steroids in 100 cases. A newer slotted tube system allowed for foraminoplasty and removal of osteophytes or extruded fragments. RESULTS: Good-to-excellent results were recorded in 432 of the 500 patients (86.4%). Separate analysis was made of the first 100 cases when the KTP laser was in use. CONCLUSIONS: The 2.8 mm operating channel scope produced clear visualization of annular tears, disc fragments, foraminal osteophytes, and the epidural space. Monitoring of the microinstruments in the disc space and spinal canal was readily accomplished. The quality of the imaging provided by discography improved the definition of the disc pathology. Adjuvant use of lasers, radiofrequency, chymopapain, and intradiscal steroids and the newer slotted tube system, have contributed to the advances in minimally invasive technique for endoscopic discectomy. PMID- 11021784 TI - Continuing experience with subgaleal shunting. AB - OBJECTIVE: To evaluate subgaleal shunting--the diversion of cerebrospinal fluid into the subgaleal space for temporary absorption by the membranes of the scalp. METHOD: Eighty-one (81) patients were treated over a 20-year period. There were 22 cases of hypertensive hydrocephalus, 52 cases of acute head trauma, and 7 cases of chronic subdural hematoma. The simple surgical technique is described. RESULTS: Of the eight-one patients, 33 improved, 27 did not change, and 21 expired within 72 hours. Intracranial pressure was monitored directly in 22 cases and demonstrated that subgaleal shunting provided effective short-term treatment. CONCLUSIONS: This simple, closed method for drainage of cerebrospinal fluid avoids the complications of open ventriculostomy or open drainage of the subdural space. PMID- 11021786 TI - Supraorbital burr hole via a glabellar incision: a simple approach to anterior skull base. AB - A left subfrontopolar lesion with marked edema was totally resected utilizing a minimally invasive approach. It was possible to expose and resect the lesion, which turned out to be a tuberculoma, through a burr hole placed supraorbitally through a glabellar incision. The development and significance of minimalization techniques for surgery in the skull base region are discussed. PMID- 11021787 TI - Herpes zoster. PMID- 11021788 TI - Discovery of young, isolated planetary mass objects in the final sigma Orionis star cluster. AB - We present the discovery by optical and near-infrared imaging of an extremely red, low-luminosity population of isolated objects in the young, nearby stellar cluster around the multiple, massive star final sigma Orionis. The proximity (352 parsecs), youth (1 million to 5 million years), and low internal extinction make this cluster an ideal site to explore the substellar domain from the hydrogen mass limit down to a few Jupiter masses. Optical and near-infrared low-resolution spectroscopy of three of these objects confirms the very cool spectral energy distribution (atmospheric effective temperatures of 1700 to 2200 kelvin) expected for cluster members with masses in the range 5 to 15 times that of Jupiter. Like the planets of the solar system, these objects are unable to sustain stable nuclear burning in their interiors, but in contrast they are not bound to stars. This new kind of isolated giant planet, which apparently forms on time scales of less than a few million years, offers a challenge to our understanding of the formation processes of planetary mass objects. PMID- 11021789 TI - Optically defined multifunctional patterning of photosensitive thin-film silica mesophases. AB - Photosensitive films incorporating molecular photoacid generators compartmentalized within a silica-surfactant mesophase were prepared by an evaporation-induced self-assembly process. Ultraviolet exposure promoted localized acid-catalyzed siloxane condensation, which can be used for selective etching of unexposed regions; for "gray-scale" patterning of refractive index, pore size, surface area, and wetting behavior; and for optically defining a mesophase transformation (from hexagonal to tetragonal) within the film. The ability to optically define and continuously control both structure and function on the macro- and mesoscales is of interest for sensor arrays, nanoreactors, photonic and fluidic devices, and low-dielectric-constant films. PMID- 11021790 TI - Vibrational promotion of electron transfer. AB - By using laser methods to prepare specific quantum states of gas-phase nitric oxide molecules, we examined the role of vibrational motion in electron transfer to a molecule from a metal surface free from the complicating influence of solvation effects. The signature of the electron transfer process is a highly efficient multiquantum vibrational relaxation event, where the nitrogen oxide loses hundreds of kilojoules per mole of energy on a subpicosecond time scale. These results cannot be explained simply on the basis of Franck-Condon factors. The large-amplitude vibrational motion associated with molecules in high vibrational states strongly modulates the energetic driving force of the electron transfer reaction. These results show the importance of molecular vibration in promoting electron transfer reactions, a class of chemistry important to molecular electronics devices, solar energy conversion, and many biological processes. PMID- 11021791 TI - Dynamically controlled protein tunneling paths in photosynthetic reaction centers. AB - Marcus theory has explained how thermal nuclear motions modulate the energy gap between donor and acceptor sites in protein electron transfer reactions. Thermal motions, however, may also modulate electron tunneling between these reactions. Here we identify a new mechanism of nuclear dynamics amplification that plays a central role when interference among the dominant tunneling pathway tubes is destructive. In these cases, tunneling takes place in protein conformations far from equilibrium that minimize destructive interference. As an example, we demonstrate how this dynamical amplification mechanism affects certain reaction rates in the photosynthetic reaction center and therefore may be critical for biological function. PMID- 11021792 TI - Xenon as a complex ligand: the tetra xenono Gold(II) cation in AuXe(4)2+(Sb(2)F(11)-)(2). AB - The first metal-xenon compound with direct gold-xenon bonds is achieved by reduction of AuF(3) with elemental xenon. The square planar AuXe(4)2+ cation is established by a single-crystal structure determination, with a gold-xenon bond length of approximately 274 picometers. The bonding between gold and xenon is of the final sigma donor type, resulting in a charge of approximately 0.4 per xenon atom. PMID- 11021793 TI - Resurrection of crushed magnetization and chaotic dynamics in solution NMR spectroscopy. AB - We show experimentally and theoretically that two readily observed effects in solution nuclear magnetic resonance (NMR)-radiation damping and the dipolar field combine to generate bizarre spin dynamics (including chaotic evolution) even with extraordinarily simple sequences. For example, seemingly insignificant residual magnetization after a crusher gradient triggers exponential regrowth of the magnetization, followed by aperiodic turbulent spin motion. The estimated Lyapunov exponent suggests the onset of spatial-temporal chaos and the existence of chaotic attractors. This effect leads to highly irreproducible experimental decays that amplify minor nonuniformities such as temperature gradients. Imaging applications and consequences for other NMR studies are discussed. PMID- 11021794 TI - Imaging the electron wave function in self-assembled quantum dots. AB - Magnetotunneling spectroscopy is used as a noninvasive and nondestructive probe to produce two-dimensional spatial images of the probability density of an electron confined in a self-assembled semiconductor quantum dot. The technique exploits the effect of the classical Lorentz force on the motion of a tunneling electron and can be regarded as the momentum (k) space analog of scanning tunneling microscopy imaging. The images reveal the elliptical symmetry of the ground state and the characteristic lobes of the higher energy states. PMID- 11021795 TI - Potent analgesic effects of GDNF in neuropathic pain states. AB - Neuropathic pain arises as a debilitating consequence of nerve injury. The etiology of such pain is poorly understood, and existing treatment is largely ineffective. We demonstrate here that glial cell line-derived neurotrophic factor (GDNF) both prevented and reversed sensory abnormalities that developed in neuropathic pain models, without affecting pain-related behavior in normal animals. GDNF reduces ectopic discharges within sensory neurons after nerve injury. This may arise as a consequence of the reversal by GDNF of the injury induced plasticity of several sodium channel subunits. Together these findings provide a rational basis for the use of GDNF as a therapeutic treatment for neuropathic pain states. PMID- 11021796 TI - Long-term survival but impaired homeostatic proliferation of Naive T cells in the absence of p56lck. AB - Interactions between the T cell receptor (TCR) and major histocompatibility complex antigens are essential for the survival and homeostasis of peripheral T lymphocytes. However, little is known about the TCR signaling events that result from these interactions. The peripheral T cell pool of p56lck (lck)-deficient mice was reconstituted by the expression of an inducible lck transgene. Continued survival of peripheral naive T cells was observed for long periods after switching off the transgene. Adoptive transfer of T cells from these mice into T lymphopoienic hosts confirmed that T cell survival was independent of lck but revealed its essential role in TCR-driven homeostatic proliferation of naive T cells in response to the T cell-deficient host environment. These data suggest that survival and homeostatic expansion depend on different signals. PMID- 11021797 TI - Molecular and neuronal substrate for the selective attenuation of anxiety. AB - Benzodiazepine tranquilizers are used in the treatment of anxiety disorders. To identify the molecular and neuronal target mediating the anxiolytic action of benzodiazepines, we generated and analyzed two mouse lines in which the alpha2 or alpha3 GABAA (gamma-aminobutyric acid type A) receptors, respectively, were rendered insensitive to diazepam by a knock-in point mutation. The anxiolytic action of diazepam was absent in mice with the alpha2(H101R) point mutation but present in mice with the alpha3(H126R) point mutation. These findings indicate that the anxiolytic effect of benzodiazepine drugs is mediated by alpha2 GABAA receptors, which are largely expressed in the limbic system, but not by alpha3 GABAA receptors, which predominate in the reticular activating system. PMID- 11021798 TI - Function of GATA transcription factors in preadipocyte-adipocyte transition. AB - Genes that control the early stages of adipogenesis remain largely unknown. Here, we show that murine GATA-2 and GATA-3 are specifically expressed in white adipocyte precursors and that their down-regulation sets the stage for terminal differentiation. Constitutive GATA-2 and GATA-3 expression suppressed adipocyte differentiation and trapped cells at the preadipocyte stage. This effect is mediated, at least in part, through the direct suppression of peroxisome proliferator-activated receptor gamma. GATA-3-deficient embryonic stem cells exhibit an enhanced capacity to differentiate into adipocytes, and defective GATA 2 and GATA-3 expression is associated with obesity. Thus, GATA-2 and GATA-3 regulate adipocyte differentiation through molecular control of the preadipocyte adipocyte transition. PMID- 11021799 TI - Proximity of chromosomal loci that participate in radiation-induced rearrangements in human cells. AB - Rearrangements involving the RET gene are common in radiation-associated papillary thyroid cancer (PTC). The RET/PTC1 type of rearrangement is an inversion of chromosome 10 mediated by illegitimate recombination between the RET and the H4 genes, which are 30 megabases apart. Here we ask whether despite the great linear distance between them, RET and H4 recombination might be promoted by their proximity in the nucleus. We used two-color fluorescence in situ hybridization and three-dimensional microscopy to map the positions of the RET and H4 loci within interphase nuclei. At least one pair of RET and H4 was juxtaposed in 35% of normal human thyroid cells and in 21% of peripheral blood lymphocytes, but only in 6% of normal mammary epithelial cells. Spatial contiguity of RET and H4 may provide a structural basis for generation of RET/PTC1 rearrangement by allowing a single radiation track to produce a double strand break in each gene at the same site in the nucleus. PMID- 11021800 TI - A calmodulin-related protein that suppresses posttranscriptional gene silencing in plants. AB - Posttranscriptional gene silencing (PTGS) is an ancient eukaryotic regulatory mechanism in which a particular RNA sequence is targeted and destroyed. The helper component-proteinase (HC-Pro) of plant potyviruses suppresses PTGS in plants. Using a yeast two-hybrid system, we identified a calmodulin-related protein (termed rgs-CaM) that interacts with HC-Pro. Here we report that rgs-CaM, like HC-Pro itself, suppresses gene silencing. Our work is the first report identifying a cellular suppressor of PTGS. PMID- 11021801 TI - Regulation of STAT3 by direct binding to the Rac1 GTPase. AB - The signal transducers and activators of transcription (STAT) transcription factors become phosphorylated on tyrosine and translocate to the nucleus after stimulation of cells with growth factors or cytokines. We show that the Rac1 guanosine triphosphatase can bind to and regulate STAT3 activity. Dominant negative Rac1 inhibited STAT3 activation by growth factors, whereas activated Rac1 stimulated STAT3 phosphorylation on both tyrosine and serine residues. Moreover, activated Rac1 formed a complex with STAT3 in mammalian cells. Yeast two-hybrid analysis indicated that STAT3 binds directly to active but not inactive Rac1 and that the interaction occurs via the effector domain. Rac1 may serve as an alternate mechanism for targeting STAT3 to tyrosine kinase signaling complexes. PMID- 11021802 TI - Regulation of C. elegans life-span by insulinlike signaling in the nervous system. AB - An insulinlike signaling pathway controls Caenorhabditis elegans aging, metabolism, and development. Mutations in the daf-2 insulin receptor-like gene or the downstream age-1 phosphoinositide 3-kinase gene extend adult life-span by two to threefold. To identify tissues where this pathway regulates aging and metabolism, we restored daf-2 pathway signaling to only neurons, muscle, or intestine. Insulinlike signaling in neurons alone was sufficient to specify wild type life-span, but muscle or intestinal signaling was not. However, restoring daf-2 pathway signaling to muscle rescued metabolic defects, thus decoupling regulation of life-span and metabolism. These findings point to the nervous system as a central regulator of animal longevity. PMID- 11021803 TI - Structure of the protease domain of memapsin 2 (beta-secretase) complexed with inhibitor. AB - Memapsin 2 (beta-secretase) is a membrane-associated aspartic protease involved in the production of beta-amyloid peptide in Alzheimer's disease and is a major target for drug design. We determined the crystal structure of the protease domain of human memapsin 2 complexed to an eight-residue inhibitor at 1.9 angstrom resolution. The active site of memapsin 2 is more open and less hydrophobic than that of other human aspartic proteases. The subsite locations from S4 to S2' are well defined. A kink of the inhibitor chain at P2' and the change of chain direction of P3' and P4' may be mimicked to provide inhibitor selectivity. PMID- 11021804 TI - Immune inhibitory receptors. AB - With the detailed description and analysis of several inhibitory receptor systems on lymphoid and myeloid cells, a central paradigm has emerged in which the pairing of activation and inhibition is necessary to initiate, amplify, and then terminate immune responses. In some cases, the activating and inhibitory receptors recognize similar ligands, and the net outcome is determined by the relative strength of these opposing signals. The importance of this modulation is demonstrated by the sometimes fatal autoimmune disorders observed in mice with targeted disruption of inhibitory receptors. The significance of these receptors is further evidenced by the conservation of immunoreceptor tyrosine-based inhibitory motifs during their evolution. PMID- 11021805 TI - T-Independent immune response: new aspects of B cell biology. AB - Recent results emphasize the roles of T-independent antibody response in humoral defenses, for which B1 cells and marginal zone B cells are mostly responsible. We discuss how these cells are activated, migrate, and differentiate into antibody producing cells in various lymphoid tissues. Based on recent findings in each of these areas of B cell biology, we propose a possible mechanism for peripheral tolerance of autoreactive B cells at target organs. PMID- 11021806 TI - Dynamics of T lymphocyte responses: intermediates, effectors, and memory cells. AB - The immune response is initiated in organized lymphoid tissues where antigen loaded dendritic cells (DCs) encounter antigen-specific T cells. DCs function as packets of information that must be decoded by the T cell before an appropriate immune response can be mounted. We discuss how the dynamics of DC-T cell encounter and the mechanism of T cell differentiation make the decoding of this information stochastic rather than determinate. This results in the generation of both terminally differentiated effector cells and intermediates that play distinctive roles in protection, immunoregulation, and immunological memory. PMID- 11021807 TI - The ins and outs of body surface immunology. AB - Rather than being confined to the secondary lymphoid tissue of the spleen and lymph nodes, large numbers of lymphocytes are intrinsically associated with the epithelial surfaces of the body. The best studied is gut-associated lymphoid tissue, but distinct epithelium-associated lymphoid tissue also exists in the reproductive tract, the lung, and the skin. The multiple cell types and functions composing these lymphoid tissues are increasingly seen as the key to how antigens delivered to body surfaces can elicit either immunogenic or tolerogenic responses. In some instances, these responses occur purely within the local body surface tissue, yet in other cases both local and systemic responses are elicited. PMID- 11021808 TI - Prevalence of human papillomavirus DNA in different histological subtypes of cervical adenocarcinoma. AB - The prevalence of human papilloma virus (HPV) DNA in different histological subtypes of cervical adenocarcinoma and related tumors was examined using formalin-fixed, paraffin-embedded tissue samples from 105 primary cervical adenocarcinomas and adenosquamous carcinomas. Broad-spectrum HPV DNA amplification and genotyping was performed with the SPF10 primer set and line probe assay (LiPA), respectively. HPV DNA was detected in 82 of 90 (91%) mucinous adenocarcinomas, encompassing endocervical, intestinal, and endometrioid histological subtypes, and in nine of nine adenosquamous tumors (100%). HPV DNA was not detected in any nonmucinous adenocarcinomas including clear cell, serous, and mesonephric carcinomas (0/6). The most common viral types detected in adenocarcinoma were HPV 16 (50%) and HPV 18 (40%), followed by HPV 45 (10%), HPV52 (2%), and HPV 35 (1%). Multiple HPV types were detected in 9.7% of the cases. In conclusion, mucinous adenocarcinomas and adenosquamous carcinomas of the cervix demonstrate a very high prevalence of HPV DNA, similar to that reported for cervical squamous cell carcinoma. Only rare histological variants of cervical adenocarcinoma seem unrelated to HPV infection. PMID- 11021809 TI - Histological and immunoglobulin VH gene analysis of interfollicular small lymphocytic lymphoma provides evidence for two types. AB - Interfollicular small lymphocytic lymphoma (I-SLL) has not been well characterized and its relationship to small lymphocytic lymphoma (SLL) or chronic lymphocytic leukemia (CLL) is uncertain. Moreover, two different proliferation center growth patterns have been described with respect to reactive germinal centers. In this study, we evaluate the histological and immunophenotypic features of 13 cases of I-SLL and immunoglobulin heavy chain variable (VH) gene sequences from 10 cases. Immunophenotypic analyses indicate that cases showing either growth pattern have the same CD5-positive B cell phenotype typical of SLL or CLL. Sequence analysis revealed the use of VH, D, and J gene segments often found in CLL, although there may be more frequent use of J6. Similar to recent studies of CLL, there were approximately equal numbers of cases with either mutated or unmutated VH genes without evidence of ongoing mutation, consistent with I-SLL having either a naive or memory B cell origin. Interestingly, the mutational status of the I-SLL VH genes seemed to correlate with the two different histological growth patterns. These studies support the proposal that I SLL represents SLL/CLL and suggest the recently proposed two types of CLL originating from either memory or naive B cells may have different histological patterns of growth in lymph nodes that show architectural preservation. PMID- 11021810 TI - Patterns of hairless (hr) gene expression in mouse hair follicle morphogenesis and cycling. AB - The hr (hairless) gene encodes a putative transcription factor with restricted expression in the skin and brain. Mutations in the hr locus cause papular atrichia in humans and complete hair loss in mice and other mammals. To further elucidate the role of hr in skin biology, and to identify potential target cells for hr regulation, we studied hr mRNA localization during hair follicle (HF) morphogenesis and cycling in normal C57BL/6J mice. In situ hybridization revealed that hr expression was present in the suprabasal cell layers of the epidermis, whereas the basal and highly differentiated keratinocytes of the granular layer were hr-negative. During the early stages of HF morphogenesis, hr mRNA was detected in the developing hair peg. Later, it became concentrated in the HF infundibulum, in the HF matrix, and in the inner root sheath (IRS), whereas the dermal papilla (DP) and outer root sheath were consistently hr mRNA-negative. During catagen, hr gene expression gradually declined in the regressing IRS, shortly but dramatically increased in the zone of developing club hair, and became up-regulated in the epithelial cells adjacent to the DP. The co localization of hr mRNA with the site of the morphological defects in mutant skin implicates hr as a key factor in regulating basic cellular processes during catagen, including club hair formation, maintenance of DP-epithelial integrity, IRS disintegration, and keratinocyte apoptosis in the HF matrix. PMID- 11021811 TI - Enhancement of chemical hepatocarcinogenesis by the HIV-1 tat gene. AB - The human immunodeficiency virus-1 Tat protein is suspected to be involved in the neoplastic pathology arising in AIDS patients. tat-transgenic (TT) mice, which constitutively express Tat in the liver, develop liver cell dysplasia (LCD) that may represent a preneoplastic lesion. To test if TT mice are predisposed to liver carcinogenesis, we treated them with diethylnitrosamine, a hepatotropic carcinogen. Diethylnitrosamine-treated TT mice developed both preneoplastic and neoplastic lesions in the liver. They showed an enhancement of LCD and developed basophilic liver cell nodules (BLCN), hepatocellular adenomas (HA), and hepatocellular carcinomas (HC). Both preneoplastic (LCD and BLCN) and neoplastic (HA and HC) lesions were significantly more frequent in TT than in control mice: 29.7% versus 12.7% for LCD, 57.9% versus 23.3% for BLCN, 40.6% versus 10.0% for HA, and 50.0% versus 12.7% for HC. These results indicate that Tat expression in the liver predisposes to both initiation of hepatocarcinogenesis and to malignant progression of liver tumors. This study supports a role for Tat in enhancing the effect of endogenous and exogenous carcinogens in human immunodeficiency virus-1 infected patients, thereby contributing to tumorigenesis in the course of AIDS. PMID- 11021812 TI - Mutations in exons 9 and 13 of KIT gene are rare events in gastrointestinal stromal tumors. A study of 200 cases. AB - Gastrointestinal stromal tumors (GISTs), the most common mesenchymal tumors of the gastrointestinal tract, typically express the KIT protein. Activating mutations in the juxtamembrane domain (exon 11) of the c-kit gene have been shown in a subset of GISTs. These mutations lead into ligand-independent activation of the tyrosine kinase of c-kit, and have a transforming effect in vitro. Several groups have studied the clinical implication of the c-kit mutation status of exon 11 in GISTs and a possible relationship between c-kit mutations and malignant behavior has been established. Recently, a 1530ins6 mutation in exon 9 and missense mutations, 1945A>G in exon 13 of the c-kit gene were reported. The frequency and clinical importance of these findings are unknown. In this study we evaluated 200 GISTs for the presence of mutations in exons 9 and 13 of c-kit. Six cases revealed 1530ins6 mutation in exon 9 and two cases 1945A>G mutation in exon 13. All tumors with mutations in exon 9 and 13 lacked mutations in exon 11 of c kit. None of the analyzed tumors had more than one type of c-kit mutation. All but one of the eight tumors with mutations in exon 9 or 13 of the c-kit gene were histologically and clinically malignant. All four of six cases with exon 9 mutation of which location of primary tumor was known, were small intestinal, suggesting that this type of mutation could preferentially occur in small intestinal tumors. Exon 9 and 13 mutations seem to be rare, and they cover only a small portion (8%) of the balance of GISTs that do not have mutations in exon 11 of c-kit. This finding indicates that other genetic alterations may activate c kit in GISTs, or that KIT is not activated by mutations in all cases. PMID- 11021813 TI - Mutation and expression analyses reveal differential subcellular compartmentalization of PTEN in endocrine pancreatic tumors compared to normal islet cells. AB - The pathogenesis of sporadic endocrine pancreatic tumors (EPTs) is still primarily unknown. Comparative genomic hybridization studies revealed loss of 10q in a significant number (nine of 31) of EPTs. The tumor suppressor gene PTEN lies on 10q23, and so, is a candidate to play some role in EPT pathogenesis. Germline PTEN mutations are found in Cowden and Bannayan-Riley-Ruvalcaba syndromes, whereas somatic mutations and deletions are found in a variety of sporadic cancers. The mutation and expression status of PTEN in EPTs has not yet been examined. Mutation analysis of the entire coding region of PTEN including splice sites was performed in 33 tumors, revealing one tumor with somatic L182F (exon 6). Loss of heterozygosity of the 10q23 region was detected in eight of 15 informative malignant (53%) and in none of seven benign EPTs. PTEN expression was assessed in 24 available EPTs by immunohistochemistry using a monoclonal anti PTEN antibody. Of these 24, 23 tumors showed strong immunoreactivity for PTEN. Only the EPTs with PTEN mutation lacked PTEN protein expression. Although normal islet cells always exhibited predominantly nuclear PTEN immunostaining, 19 of 23 EPTs had a predominantly cytoplasmic PTEN expression pattern. Exocrine pancreatic tissue was PTEN-negative throughout. PTEN mutation is a rare event in malignant EPTs and PTEN protein is expressed in most (23 of 24) EPTs. Thus, intragenic mutation or another means of physical loss of PTEN is rarely involved in the pathogenesis of EPTs. Instead, either an impaired transport system of PTEN to the nucleus or some other means of differential compartmentalization could account for impaired PTEN function. Loss of heterozygosity of the 10q23 region is a frequent event in malignant EPTs and might suggest several hypotheses: a different tumor suppressor gene in the vicinity of PTEN might be principally involved in EPT formation; alternatively, 10q loss, including PTEN, seems to be associated with malignant transformation, but the first step toward neoplasia might involve altered subcellular localization of PTEN. PMID- 11021814 TI - Loss of Dpc4 expression in colonic adenocarcinomas correlates with the presence of metastatic disease. AB - DPC4 is a candidate tumor suppressor gene on chromosome 18q21, a region that shows high frequencies of allelic losses in pancreatic and colorectal adenocarcinomas. Biallelic inactivation of DPC4 has been reported in half of pancreatic cancers, but are relatively infrequent in other tumor types. The role of DPC4 inactivation in colorectal neoplasms has not been fully characterized. An immunohistochemical assay for Dpc4 protein expression has been recently developed and shown to be a sensitive and specific surrogate for alterations in the DPC4 gene. In this study we examined the expression of Dpc4 protein in formalin-fixed archival tissue from 83 colorectal lesions, including 19 adenomas and 64 sporadic adenocarcinomas (11 stage I, 13 stage II, 17 stage III, and 23 stage IV cancers). None of the adenomas or stage I adenocarcinomas showed loss of Dpc4 expression, whereas one of 13 (8%) stage II, one of 17 (6%) stage III, and five of 23 (22%) of stage IV cancers showed loss of Dpc4 expression. There was a borderline significant difference in loss of Dpc4 reactivity in colorectal tumors with distant metastasis at presentation (22%) versus primary tumors without distant metastasis (5%) (Fisher's exact test, P = 0.05; chi(2) = 0.04). Poorly differentiated histology or status of pericolonic lymph nodes did not affect Dpc4 expression. Alterations in DPC4 are involved in the progression of a subset of colorectal carcinomas, especially those that present with advanced disease. In the sequential pathogenesis of colorectal tumors, inactivation of DPC4 is likely to be a late event. PMID- 11021815 TI - Patterning and nuclear beta-catenin expression in the colonic adenoma-carcinoma sequence. Analogies with embryonic gastrulation. AB - Patterning is a process by which ordered arrangements of cells and tissue structure are attained. The term derived from developmental biology is also useful for the study of colonic carcinogenesis, in which the patterning of neoplastic tubules is necessary for properties of growth, invasion, and metastasis. Interestingly the nuclear expression and transcriptional activity of beta-catenin, a major oncoprotein in colonic carcinogenesis, is decisive for the first patterning of a tubule in embryogenesis, which creates the primitive gut and is called the gastrulation. Thus, basic patternings of embryogenesis and carcinogenesis might be linked. To test this hypothesis we compared morphological patterns and immunohistochemical beta-catenin stainings in colonic adenomas and adenocarcinomas with the gastrulation steps. Two analogies were found: 1) the patterning of invasion with reconstruction in adenocarcinomas corresponded to the epithelio-mesenchymal transition, ingression, and rearrangement of cells during the first phase of gastrulation; and 2) the patterning of tubular branching in adenomas and adenocarcinomas resembled the endodermal invagination during the second phase. The intratumorous distribution and intensity of nuclear beta catenin expression was significantly correlated with the two patternings, similar to the findings in gastrulation. The results indicate microenvironmental regulations of nuclear beta-catenin expression and a return of neoplastic cells to embryonic transcriptional susceptibilities during colonic carcinogenesis. PMID- 11021816 TI - Epigenetic PTEN silencing in malignant melanomas without PTEN mutation. AB - A tumor suppressor gene at 10q 23.3, designated PTEN, encoding a dual specificity phosphatase with lipid and protein phosphatase activity, has been shown to play an important role in the pathogenesis of a variety of human cancers. Germline mutations in PTEN cause Cowden syndrome (CS), which is characterized by multiple hamartomas and a high risk of breast and thyroid cancers. Frequent loss of heterozygosity at 10q is found in both early and advanced-stage sporadic melanomas; however, mutations or deletions in PTEN are detected mainly in melanoma cell lines. In this study, we examined PTEN expression in 34 unselected sporadic melanomas (4 primary melanomas, 30 metastases) using immunohistochemistry and correlated this with the results of structural studies of this gene. Immunostaining of 34 melanoma samples revealed no PTEN expression in 5 (15%) and low PTEN expression in 17 (50%), whereas the rest of the tumors (35%) had high levels of expression. Hemizygous deletion was found in 32% of the tumors but neither intragenic PTEN mutation nor biallelic deletion was found in any of the samples. Of the 5 melanomas showing no PTEN expression, 4 had no mutation or deletion of PTEN. Of the 13 tumors having weak PTEN immunoreactivity and informative loss of heterozygosity results, 6 had evidence of hemizygous allelic loss of PTEN while the remaining 7 had intact PTEN. These results strongly support PTEN as a major tumor suppressor on 10q involved in melanoma tumorigenesis and suggest an epigenetic mechanism of biallelic functional inactivation not previously observed in other cancers where PTEN might be involved. PMID- 11021817 TI - The role of microsatellite instability in gastric low- and high-grade lymphoma development. AB - DNA mismatch repair genes and their dysfunction as evidenced by microsatellite instability (MSI) play an important role in the pathogenesis of a variety of tumors, most prominently hereditary nonpolyposis colorectal cancer (HNPCC). However, their role in development of extranodal lymphomas has not been established yet. We therefore evaluated for MSI 25 gastric low-grade marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue type and 31 gastric high-grade diffuse large B-cell lymphomas (DLBCLs) with 29 and 118 microsatellites, respectively. Compared with HNPCC, the overall level of MSI was much lower with a mean of 2.6% MSI-positive repeats in the DLBCLs; the frequency of MSI showed a tendency to increase with age (P = 0.01), as did MSI variability (P = 0.02). Low-grade mucosa-associated lymphoid tissue lymphomas displayed even less MSI (sevenfold) than DLBCLs (P = 0.009). MSI frequency thus increases with the transition from low- to high-grade disease and with age; it does not seem to initiate lymphomagenesis. Other microsatellites than those typically mutated in HNPCC frequently revealed MSI in these lymphomas, especially dinucleotide repeats on chromosomes 3, 5, and 18. To facilitate rapid screening of lymphomas for MSI and to establish a tool for future MSI frequency comparisons, we recommend to use repeats D3S1261, D3S1530, D5S346, D17S250, D18S474, and DCC. PMID- 11021818 TI - Lack of phosphotyrosine phosphatase SHP-1 expression in malignant T-cell lymphoma cells results from methylation of the SHP-1 promoter. AB - SHP-1 is an important negative regulator of signaling by several receptors including receptors for interleukin-2 (IL-2R) and other cytokines. SHP-1 acts by dephosphorylating the receptors and receptor-associated kinases such as IL-2R associated Jak3 kinase. We found that SHP-1 protein was not detectable or greatly diminished in most (six of seven) T cell lines derived from various types of T cell lymphomas and all (eight of eight) cutaneous T-cell lymphoma tissues with a transformed, large-cell morphology. All T-cell lymphoma lines tested (eight of eight) expressed diminished amounts or no detectable SHP-1 mRNA. These T cell lines did not, however, carry any mutations in the SHP-1 gene-coding, splice junction, and promoter regions. Importantly, SHP-1 DNA promoter region in the T cell lines was resistant to digestion with three different methylation-sensitive restriction enzymes. This resistance was reversed by treatment of the cells with a demethylating agent, 5-deoxyazacytidine. The treatment resulted also in the expression of SHP-1 mRNA and, less frequently, SHP-1 protein. The expression of SHP-1 protein was associated with dephosphorylation of the Jak3 kinase. These results show that lack of SHP-1 expression is frequent in malignant T cells and results from methylation of the SHP-1 gene promoter. Furthermore, they indicate that SHP-1 loss may play a role in the pathogenesis of T cell lymphomas by permitting persistence of signals generated by IL-2R and, possibly, other receptor complexes. PMID- 11021819 TI - BCL10 expression in normal and neoplastic lymphoid tissue. Nuclear localization in MALT lymphoma. AB - BCL10 is an apoptotic regulatory molecule identified through its direct involvement in t(1;14)(p22;q32) of mucosa-associated lymphoid tissue (MALT) lymphoma. We examined BCL10 protein expression in various normal tissues and B cell lymphomas by immunohistochemistry of formalin-fixed and paraffin-embedded tissues using mouse BCL10 monoclonal antibodies. BCL10 protein was expressed in lymphoid tissue but not in 21 various other tissues with the exception of breast. In normal B-cell follicles, the protein was expressed abundantly in the germinal center B cells, moderately in the marginal zone, but only weakly in the mantle zone B cells. Irrespective of their stage of B-cell maturation, BCL10 was predominantly expressed in the cytoplasm. In contrast, each of the four MALT lymphomas with t(1;14)(p22;q32) showed strong BCL10 expression in both the nucleus and cytoplasm. Twenty of 36 (55%) MALT lymphomas lacking the translocation exhibited BCL10 expression in both the nucleus and cytoplasm although at a much lower level, whereas the remaining 16 cases displayed only cytoplasmic BCL10. Unlike MALT lymphoma, both follicular and mantle cell lymphomas generally displayed BCL10 expression compatible to their normal cell counterparts. Our results show differential expression of BCL10 protein among various B-cell populations of the B-cell follicle, indicating its importance in B cell maturation. The subcellular localization of BCL10 was frequently altered in MALT lymphoma in comparison with its normal cell counterparts, suggesting that ectopic BCL10 expression may be important in the development of this type of tumor. PMID- 11021820 TI - Restricted 12p amplification and RAS mutation in human germ cell tumors of the adult testis. AB - Human testicular germ-cell tumors of young adults (TGCTs), both seminomas and nonseminomas, are characterized by 12p overrepresentation, mostly as isochromosomes, of which the biological and clinical significance is still unclear. A limited number of TGCTs has been identified with an additional high level amplification of a restricted region of 12p including the K-RAS proto oncogene. Here we show that the incidence of these restricted 12p amplifications is approximately 8% in primary TGCTs. Within a single cell formation of i(12p) and restricted 12p amplification is mutually exclusive. The borders of the amplicons cluster in short regions, and the amplicon was never found in the adjacent carcinoma in situ cells. Seminomas with the restricted 12p amplification virtually lacked apoptosis and the tumor cells showed prolonged in vitro survival like seminoma cells with a mutated RAS gene. However, no differences in proliferation index between these different groups of seminomas were found. Although patients with a seminoma containing a homogeneous restricted 12p amplification presented at a significantly younger age than those lacking it, the presence of a restricted 12p amplification/RAS mutation did not predict the stage of the disease at clinical presentation and the treatment response of primary seminomas. In 55 primary and metastatic tumors from 44 different patients who failed cisplatinum-based chemotherapy, the restricted 12p amplification and RAS mutations had the same incidence as in the consecutive series of responding patients. These data support the model that gain of 12p in TGCTs is related to invasive growth. It allows tumor cells, in particular those showing characteristics of early germ cells (ie, the seminoma cells), to survive outside their specific microenvironment. Overexpression of certain genes on 12p probably inhibits apoptosis in these tumor cells. However, the copy numbers of the restricted amplification of 12p and K-RAS mutations do not predict response to therapy and survival of the patients. PMID- 11021821 TI - Expression of heparanase in normal, dysplastic, and neoplastic human colonic mucosa and stroma. Evidence for its role in colonic tumorigenesis. AB - The human heparanase gene, an endo-beta-glucuronidase that cleaves heparan sulfate at specific intrachain sites, has recently been cloned and shown to function in tumor progression and metastatic spread. Antisense digoxigenin labeled heparanase RNA probe and monoclonal anti-human heparanase antibodies were used to examine the expression of the heparanase gene and protein in normal, dysplastic, and neoplastic human colonic mucosa. To our knowledge, this is the first systematic study of heparanase expression in human colon cancer. Both the heparanase gene and protein were expressed at early stages of neoplasia, already at the stage of adenoma, but were practically not detected in the adjacent normal looking colon epithelium. Gradually increasing expression of heparanase was evident as the cells progressed from severe dysplasia through well-differentiated to poorly differentiated colon carcinoma. Deeply invading colon carcinoma cells showed the highest levels of the heparanase mRNA and protein associated with expression of both the gene and enzyme by adjacent desmoplastic stromal fibroblasts. A high expression was also found in colon carcinoma metastases to lung, liver, and lymph nodes, as well as in the accompanying stromal fibroblasts. Moreover, extracts derived from tumor tissue expressed much higher levels of the heparanase protein and activity as compared to the normal colon tissue. In all specimens, the heparanase gene and protein exhibited the same pattern of expression. These results suggest a role of heparanase in colon cancer progression and may have both prognostic and therapeutic applications. PMID- 11021822 TI - Novel protective effects of stem cell factor in a murine model of acute septic peritonitis. Dependence on MCP-1. AB - Mast cells participate in the host response during sepsis and have been shown to have a protective effect in a murine model of acute septic peritonitis and multi organ failure initiated by cecal ligation and puncture (CLP). Stem cell factor (SCF) is a hematopoietic cytokine important in mast cell proliferation and activation. In the present study, we examined the protective effects of a single intraperitoneal injection of SCF given 2 hours before CLP surgery in mice. Four days after the CLP surgery, SCF pretreatment significantly improved mouse survival from 29 to 56% and mast cells were absolutely required for this effect. Immunoneutralization studies revealed that the SCF-stimulated release of monocyte chemoattractant protein-1 (MCP-1) into the septic peritoneal cavity contributed to the protective effect of SCF in this model. One potential cellular source of MCP-1 was the SCF-activated mast cell. In addition, SCF pretreatment significantly augmented circulating levels of SCF and the immunomodulatory cytokine interleukin-10 in septic mice, in part because the SCF pretreatment seemed to promote the release of both mediators from the liver. Additional hepatic effects of SCF treatment included an accelerated expression of hepatic levels of signal transducer and activator of transcription-3 (STAT-3) in CLP mice pretreated with SCF. Taken together, the findings from the present study demonstrate that the intraperitoneal delivery of SCF has a major protective effect in a murine model of CLP. PMID- 11021824 TI - Expression of intrinsic factor in rat and murine gastric mucosal cell lineages is modified by inflammation. AB - Intrinsic factor is produced primarily by chief cells in rat and mouse, but 4 to 11% of isolated rat parietal cells also contain intrinsic factor. To test whether local conditions could alter the distribution of intrinsic factor expression, two rodent models of chronic lymphocytic gastric inflammation were examined. Immunocytochemistry was performed using antiserum against human intrinsic factor and H/K ATPase (a parietal cell marker), counting the percent of intrinsic factor positive parietal cells. HLA-B27 transgenic rats develop chronic gastritis at age 3 months. Congenic controls expressed intrinsic factor in 8.9 +/- 3.8% (mean +/- SD) of parietal cells; in inflamed areas of transgenic rats 21 +/- 5.2% (P < 0.0001) of parietal cells were positive. In adjacent areas without inflammatory infiltrate 16 +/- 3.6% of parietal cells contained intrinsic factor. C57BL/6 mice inoculated with Helicobacter felis develop gastritis by 4 weeks. After 4 and 8 weeks of infection, intrinsic factor-positive parietal cells increased from 7.8 +/- 2.8% in the congenic controls to 17.6 +/- 4.1% in the inflamed gastric body (P < 0.0001). Isolated rat parietal cells incubated with interleukin-1beta demonstrated a twofold increase in intrinsic factor-positive parietal cells. These studies are consistent with the concept that intrinsic factor expression is both predetermined in chief cells and can be expressed in parietal cells in response to local inflammatory factors. The differences between inflamed and adjacent noninflamed areas in the rat model suggest a tissue gradient of soluble inducer(s), possibly cytokines. PMID- 11021823 TI - Murine oncostatin M stimulates mouse synovial fibroblasts in vitro and induces inflammation and destruction in mouse joints in vivo. AB - Oncostatin M (OSM) is a multifunctional cytokine, a member of the interleukin 6/leukemia inhibitory factor (IL-6/LIF) family, that can regulate a number of connective-tissue cell types in vitro including cartilage and synovial tissue derived fibroblasts, however its role in joint inflammation in vivo is not clear. We have analyzed murine OSM (muOSM) activity in vitro and in vivo in mouse joint tissue, to determine the potential role of this cytokine in local joint inflammation and pathology. The effects of muOSM and other IL-6/LIF cytokines on mouse synovial fibroblast cultures were assessed in vitro and showed induction of monocyte chemotactic protein-1, interleukin-6, and tissue inhibitor metalloproteinase-1, as well as enhancement of colony growth in soft agarose culture. Other IL-6/LIF cytokines including IL-6, LIF, or cardiotrophin-1, did not have such effects when tested at relatively high concentrations (20 ng/ml). To assess effects of muOSM in articular joints in vivo, we used recombinant adenovirus expressing muOSM cDNA (AdmuOSM) and injected purified recombinant virus (10(6) to 10(8) pfu) intra-articularly into the knees of various mouse strains. Histological analysis revealed dramatic alterations in the synovium but not in synovium of knees treated with the control virus Ad-dl70 or knees treated with Adm-IL-6 encoding biologically active murine IL-6. AdmuOSM effects were characterized by increases in the synovial cell proliferation, infiltration of mononuclear cells, and increases in extracellular matrix deposition that were evident at day 4, but much more marked at days 7, 14, and 21 after administration. The synovium took on characteristics similar to pannus and appeared to contact and invade cartilage. Collectively, these results provide good evidence that OSM regulates synovial fibroblast function differently than other IL-6-type cytokines, and can induce a proliferative invasive phenotype of synovium in vivo in mice on overexpression. We suggest that OSM may contribute to pathology in arthritis. PMID- 11021825 TI - Fibronectin-mononuclear cell interactions regulate type 1 helper T cell cytokine network in tolerant transplant recipients. AB - Fibronectin (FN), expressed primarily by macrophages, endothelial cells, and smooth muscle cells, represents an integral feature of the rejection response in transplant recipients. Here we demonstrate a unique pattern of cellular FN expression in rat recipients of cardiac allografts rendered tolerant in an infectious manner with either nondepleting CD4 mAb or regulatory spleen cells. Unlike in rejecting controls, cellular FN in tolerant hosts was restricted to the graft vessels and no vascular cell adhesion molecule-1 or intercellular adhesion molecule-1 expression could be found, supporting the role of FN in leukocyte sequestration at the graft site. The lack of myocardial FN in tolerant rats, despite dense macrophage infiltration, correlated with profound depression of Th1 (interleukin-2 and interferon-gamma) cytokines. Treatment with CD4-depleting mAb prevented tolerance induction and restored myocardial expression of FN in parallel with marked increase in the expression of interleukin-2 and interferon gamma mRNA/protein. Furthermore, connective segment-1 peptide-facilitated adjunctive blockade of FN-alpha4beta1 interactions in recipients conditioned with CD4 depleting mAb, significantly depressed intragraft expression of interleukin-2 and interferon-gamma mRNA/protein. Hence, the lack of FN associated with infiltrating leukocytes plays an important role in the maintenance of tolerance in transplant recipients by depressing local expression of Th1 cytokines that otherwise facilitate acute graft rejection. PMID- 11021826 TI - Immunohistochemical expression of uPA, uPAR, and PAI-1 in breast carcinoma. Fibroblastic expression has strong associations with tumor pathology. AB - The urokinase-type plasminogen activator (uPA) system has been implicated in tumor spread. We have used immunohistochemistry to examine three components of this system, ie, uPA, uPA receptor (uPAR), and plasminogen activator inhibitor-1 (PAI-1), in a pilot study on 142 cases of breast carcinoma. We wished to determine whether there were any relationships between expression of the proteins in either tumor cells or fibroblasts and clinical and pathological features. Strong uPA expression in each cell type was significantly related to high tumor grade (P = 0.013 and 0.008, respectively), and was more common in invasive than in in situ carcinomas (P < 0.0001). Fibroblastic expression of uPAR was only related to the presence of invasion (P < 0.0001). Strong PAI-1 expression in both cell types was seen in high-grade tumors (tumor cells, P = 0.012; fibroblasts, P < 0.001), but only fibroblastic expression was related to the presence of invasion (P = 0.042). Fibroblastic expression of both uPA and uPAR were positively correlated with tumor size. Although patients with strong fibroblastic expression of uPA showed a tendency toward a shorter time to relapse, none of the plasminogen activator proteins were significantly associated with relapse-free survival. These results suggest that strong expression of uPA, uPAR, and PAI-1 in fibroblasts rather than in tumor cells have the most impact on the clinical behavior of breast cancer. Larger prospective studies are needed to confirm these findings. PMID- 11021827 TI - Induction of vascular adhesion protein-1 during liver allograft rejection and concomitant cytomegalovirus infection in rats. AB - Vascular adhesion protein-1 (VAP-1) is an adhesion molecule controlling lymphocyte recirculation through high endothelial venules of the lymph nodes. It has also been shown to be induced and to mediate lymphocyte adhesion at sites of inflammation. We studied the expression of VAP-1 and two other inducible adhesion molecules ICAM-1 and VCAM-1 in our experimental model of rat liver allograft rejection and, in addition, the effect of concomitant rat cytomegalovirus (RCMV) infection on this expression. Expression of VAP-1, ICAM-1, and VCAM-1 was studied in rat liver allografts with or without RCMV infection, isografts, and normal rat liver. Immunoperoxidase technique and monoclonal antibodies including a novel anti-VAP-1 reagent were used. VAP-1 expression was induced by acute rejection in sinusoids, hepatocytes, and also in bile ducts, when compared to the isografts or normal liver, where only blood vessels were consistently positive. Sinusoidal and hepatocyte expression of VAP-1 was prolonged by the presence of RCMV. ICAM-1 and VCAM-1 expression was also induced by acute rejection. However, RCMV increased sinusoidal VCAM-1 expression compared to uninfected grafts. The present experimental study shows that VAP-1 is up-regulated in acute rejection of liver allografts, and that this up-regulation is prolonged by RCMV infection. PMID- 11021828 TI - Antibody-mediated resolution of light chain-associated amyloid deposits. AB - Primary light-chain-associated (AL) amyloidosis is characterized by the deposition in tissue of monoclonal light chains as fibrils. With rare exception, this process is seemingly irreversible and results in progressive organ dysfunction and eventually death. To determine whether immune factors can effect amyloid removal, we developed an experimental model in which mice were injected with amyloid proteins extracted from the spleens or livers of patients with AL amyloidosis. Notably, the resultant amyloidomas were rapidly resolved, as compared to controls, when animals received injections of an anti-light-chain monoclonal antibody having specificity for an amyloid-related epitope. The reactivity of this monoclonal antibody was not dependent on the V(L) or C(L) isotype of the fibril, but rather seemed to be directed toward a beta-pleated sheet conformational epitope expressed by AL and other amyloid proteins. The amyloidolytic response was associated with a pronounced infiltration of the amyloidoma with neutrophils and putatively involved opsonization of fibrils by the antibody, leading to cellular activation and release of proteolytic factors. The demonstration that AL amyloid resolution can be induced by passive administration of an amyloid-reactive antibody has potential clinical benefit in the treatment of patients with primary amyloidosis and other acquired or inherited amyloid-associated disorders. PMID- 11021830 TI - Localization of apoptotic macrophages at the site of plaque rupture in sudden coronary death. AB - Although apoptosis is a well-recognized phenomenon in chronic atherosclerotic disease, its role in sudden coronary death, in particular, acute plaque rupture is unknown. Culprit lesions from 40 cases of sudden coronary death were evaluated. Cases were divided into two mechanisms of death: ruptured plaques with acute thrombosis (n = 25) and stable plaques with and without healed myocardial infarction (n = 15). Apoptotic cells were identified by staining of fragmented DNA and confirmed in select cases by gold conjugate labeling combined with ultrastructural analysis. Additional studies were performed to examine the expression and activation of two inducers of apoptosis, caspases-1 and -3. Ruptured plaques showed extensive macrophage infiltration of the fibrous cap, in particular at rupture sites contrary to stable lesions, which contained fewer inflammatory cells. Among the culprit lesions, the overall incidence of apoptosis in fibrous caps was significantly greater in ruptured plaques (P < 0.001) and was predominantly localized to the CD68-positive macrophages. Furthermore, apoptosis at plaque rupture sites was more frequent than in areas of intact fibrous cap (P = 0. 028). Plaque rupture sites demonstrated a strong immunoreactivity to caspase 1 within the apoptotic macrophages; staining for caspase-3 was weak. Immunoblot analysis of ruptured plaques demonstrated caspase-1 up-regulation and the presence of its active p20 subunit whereas stable lesions showed only the precursor; nonatherosclerotic control segments were negative for both precursor and active enzyme. These findings demonstrate extensive apoptosis of macrophages limited to the site of plaque rupture. The proteolytic cleavage of caspase-1 in ruptured plaques suggests activation of this apoptotic precursor. Whether macrophage apoptosis is essential to acute plaque rupture or is a response to the rupture itself remains to be determined. PMID- 11021829 TI - p75(NTR) mediates neurotrophin-induced apoptosis of vascular smooth muscle cells. AB - The development of atherosclerotic lesions results from aberrant cell migration, proliferation, and extracellular matrix production. In advanced lesions, however, cellular apoptosis, leading to lesion remodeling, predominates. During lesion formation, the neurotrophins and the neurotrophin receptor tyrosine kinases, trks B and C, are induced and mediate smooth muscle cell migration. Here we demonstrate that a second neurotrophin receptor, p75(NTR), is expressed by established human atherosclerotic lesions and late lesions that develop after balloon injury of the rat thoracic aorta. The p75(NTR), a member of the tumor necrosis factor/FAS receptor family, can modulate trk receptor function as well as initiate cell death when expressed in cells of the nervous system that lack kinase-active trk receptors. p75(NTR) expression colocalizes to neointimal cells, which express smooth muscle cell alpha-actin and are expressed by cultured human endarterectomy-derived cells (HEDC). Areas of the plaque expressing p75(NTR) demonstrate increased TUNEL positivity, and HEDC undergo apoptosis in response to the neurotrophins. Finally, neurotrophins also induced apoptosis of a smooth muscle cell line genetically manipulated to express p75(NTR), but lacking trk receptor expression. These studies identify the regulated expression of neurotrophins and p75(NTR) as an inducer of smooth muscle cell apoptosis in atherosclerotic lesions. PMID- 11021831 TI - Mechanism of abnormal elastin gene expression in the pinguecular part of pterygia. AB - Elastodysplasia and elastodystrophy are two known manifestations in the conjunctival, ie, pinguecular, part of pterygia. But the mechanisms are still not understood. The purpose of this study is to investigate the mechanism of enhanced elastin gene expression in fibroblasts from the pinguecular part of pterygia, which is related to abnormal elastic fiber expression in the pinguecular part of pterygia. Elastin in surgical specimens of normal conjunctiva and the pinguecular part of pterygia from age-matched patients was detected by immunohistochemical staining. Northern hybridization and quantification of radiolabeled tropoelastin were performed in conjunctival fibroblasts cultured under different doses of ultraviolet (UV) B irradiation, and in cultured pinguecular fibroblasts from pterygia. In vitro translation was also performed to analyze the tropoelastin production in rabbit reticulocyte lysate. The level of tropoelastin in reticulolysates from UV-treated conjunctival and pinguecular fibroblasts of pterygia was higher than in normal conjunctival fibroblasts. The coding sequence and 3'- untranslated region of tropoelastin mRNAs were amplified by reverse transcription-polymerase chain reaction, and mutations were checked by DNA sequencing. Immunohistochemical staining revealed elastin in pinguecular subepithelial connective tissues of pterygia, but not in normal conjunctiva. Tropoelastin mRNA levels were not elevated in cultured pinguecular or conjunctival fibroblasts with or without ultraviolet B irradiation. However, tropoelastin synthesis was enhanced in culture medium of pinguecular and UV irradiated conjunctival fibroblasts, but not in normal conjunctival fibroblasts. Direct DNA sequencing revealed mutations in the 3'-untranslated region but not in the coding sequence of tropoelastin mRNA, in both pinguecular and UV-irradiated conjunctival fibroblasts. The increased expression of tropoelastin in pinguecular and UV-irradiated fibroblasts is not a result of increased levels of steady-state mRNA, but is a result of posttranscriptional modification of tropoelastin. PMID- 11021832 TI - CD18 and ICAM-1-dependent corneal neovascularization and inflammation after limbal injury. AB - Extensive limbal injury is a leading cause of irreversible blindness. The destruction of corneal limbal stem cells often results in corneal neovascularization and an optically inferior epithelium. Previous work has shown that the neovascularization after limbal injury is vascular endothelial growth factor (VEGF)-dependent, with much of the VEGF emanating from the inflammatory cells that invade the cornea. Using a relevant mouse model of limbal injury, we examined the role of CD18 and intercellular adhesion molecule-1 (ICAM-1) in limbal injury-induced neovascularization. The results show that CD18- and ICAM-1 deficient mice developed 35% (n = 5, P = 0.003) and 36% (n = 5, P = 0.002) less neovascularization than strain-specific normal controls, respectively. The corneal neutrophil counts were similarly reduced by 51% (n = 5, P < 0.003) and 46% (n = 5, P < 0.006), respectively. When VEGF mRNA levels were analyzed, they were reduced by 66% (n = 3, P = 0.004) and 48% (n = 3, P = 0.024), respectively. Taken together, these data identify CD-18 and ICAM-1 as mediators of the inflammatory and VEGF-dependent corneal neovascularization that follows limbal injury. The targeting of CD18 and ICAM-1 may prove useful in the treatment of inflammation-associated neovascularization in the cornea and elsewhere. PMID- 11021833 TI - Prominent cerebral amyloid angiopathy in transgenic mice overexpressing the london mutant of human APP in neurons. AB - Deposition of amyloid beta-peptide (Abeta) in cerebral vessel walls (cerebral amyloid angiopathy, CAA) is very frequent in Alzheimer's disease and occurs also as a sporadic disorder. Here, we describe significant CAA in addition to amyloid plaques, in aging APP/Ld transgenic mice overexpressing the London mutant of human amyloid precursor protein (APP) exclusively in neurons. The number of amyloid-bearing vessels increased with age, from approximately 10 to >50 per coronal brain section in APP/Ld transgenic mice, aged 13 to 24 months. Vascular amyloid was preferentially deposited in arterioles and ranged from small focal to large circumferential depositions. Ultrastructural analysis allowed us to identify specific features contributing to weakening of the vessel wall and aneurysm formation, ie, disruption of the external elastic lamina, thinning of the internal elastic lamina, interruption of the smooth muscle layer, and loss of smooth muscle cells. Biochemically, the much lower Abeta42:Abeta40 ratio evident in vascular relative to plaque amyloid, demonstrated that in blood vessel walls Abeta40 was the more abundant amyloid peptide. The exclusive neuronal origin of transgenic APP, the high levels of Abeta in cerebrospinal fluid compared to plasma, and the specific neuroanatomical localization of vascular amyloid strongly suggest specific drainage pathways, rather than local production or blood uptake of Abeta as the primary mechanism underlying CAA. The demonstration in APP/Ld mice of rare vascular amyloid deposits that immunostained only for Abeta42, suggests that, similar to senile plaque formation, Abeta42 may be the first amyloid to be deposited in the vessel walls and that it entraps the more soluble Abeta40. Its ability to diffuse for larger distances along perivascular drainage pathways would also explain the abundance of Abeta40 in vascular amyloid. Consistent with this hypothesis, incorporation of mutant presenilin-1 in APP/Ld mice, which resulted in selectively higher levels of Abeta42, caused an increase in CAA and senile plaques. This mouse model will be useful in further elucidating the pathogenesis of CAA and Alzheimer's disease, and will allow testing of diagnostic and therapeutic strategies. PMID- 11021834 TI - Synaptic vesicle protein 2, A new neuroendocrine cell marker. AB - Synaptic vesicle protein 2 (SV2) is a glycoprotein identified in the nervous system of several species, including man, but its occurrence in the human neuroendocrine (NE) cell system has not been investigated. By using a monoclonal antibody to SV2, immunoreactivities were demonstrated in NE cell types in human gastrointestinal tract, pancreas, anterior pituitary gland, thyroid, parathyroid, and adrenal medulla, and also in chief cells of gastric oxyntic mucosa. Immunoelectron microscopy of pancreatic islets revealed SV2 immunoreactivity in secretory granules. Comparison of SV2, synaptophysin, and chromogranin A immunoreactivity showed more SV2- and synaptophysin- than chromogranin A immunoreactive cells in the antrum and pancreas. In the other gastrointestinal regions and in the other endocrine organs more SV2- than synaptophysin immunoreactive cells were seen. More chromogranin A- than SV2-immunoreactive cells were observed in duodenum, colon, and parathyroid. Various NE tumors were examined and all contained SV2-immunoreactive cells. The staining patterns with the three markers agreed well, except in hindgut carcinoids, which showed strong SV2 immunoreactivity, weak synaptophysin but no chromogranin A immunostaining. In pituitary adenomas more cells were immunoreactive to SV2 than to the other two antibodies. In conclusion, SV2 is recognized as a further broad marker for NE cells and widens the arsenal of diagnostic tools for NE tumors. It is of special importance for identifying hindgut carcinoids. PMID- 11021835 TI - Expression of Egr-1 in late stage emphysema. AB - The transcription factor early growth response (Egr)-1 is an immediate-early gene product rapidly and transiently expressed after acute tissue injury. In contrast, in this report we demonstrate that lung tissue from patients undergoing lung reduction surgery for advanced emphysema, without clinical or anatomical evidence of acute infection, displays a selective and apparently sustained increase in Egr 1 transcripts and antigen, compared with a broad survey of other genes, including the transcription factor Sp1, whose levels were not significantly altered. Enhanced Egr-1 expression was especially evident in smooth muscle cells of bronchial and vascular walls, in alveolar macrophages, and some vascular endothelium. Gel shift analysis with (32)P-labeled Egr probe showed a band with nuclear extracts from emphysematous lung which was supershifted with antibody to Egr-1. Egr-1 has the capacity to regulate genes relevant to the pathophysiology of emphysema, namely those related to extracellular matrix formation and remodeling, thrombogenesis, and those encoding cytokines/chemokines and growth factors. Thus, we propose that further analysis of Egr-1, which appears to be up regulated in a sustained fashion in patients with late stage emphysema, may provide insights into the pathogenesis of this destructive pulmonary disease, as well as a new facet in the biology of Egr-1. PMID- 11021836 TI - Possible involvement of myofibroblasts in cellular recovery of uranyl acetate induced acute renal failure in rats. AB - Cellular recovery in acute renal failure is a form of wound healing. Fibroblast like cells or myofibroblasts are involved in wound healing. We examined the serial changes in tubular damage and origin and kinetics of regenerating cells in uranyl acetate-induced acute renal failure, with a special emphasis on interstitial myofibroblasts. Acute renal failure was induced in rats by intravenous injection of uranyl acetate (5 mg/kg). All rats received bromodeoxyuridine intraperitoneally 1 hour before sacrifice. Serial changes in the distribution of tubular necrosis and bromodeoxyuridine-incorporated or vimentin-positive regenerating cells, and their spatial and temporal relation to alpha-smooth muscle actin-positive myofibroblasts as well as ED 1-positive monocytes/macrophages were examined. Necrotic tubules initially appeared around the corticomedullary junction after uranyl acetate injection, then spread both downstream and upstream of proximal tubules. Peritubular alpha-smooth muscle actin-positive myofibroblasts appeared and extended along the denuded tubular basement membrane, establishing network formation throughout the cortex and the outer stripe of outer medulla at days 4 to 5. Tubular regeneration originated in nonlethally injured cells in the distal end of S3 segments, which was confirmed by lectin and immunohistochemical staining using markers for tubular segment. Subsequently, upstream proliferation was noted along the tubular basement membrane firmly attached by myofibroblasts. During cellular recovery, no entry of myofibroblasts into the tubular lumen across the tubular basement membrane was noted and only a few myofibroblasts showed bromodeoxyuridine positivity. The fractional area of alpha-smooth muscle actin-positive interstitium reached a peak level at day 7 in the cortex and outer stripe of outer medulla, then gradually disappeared by day 15 and remained only around dilated tubules and in the expanded interstitium at day 21. ED 1-positive monocytes/macrophages were transiently infiltrated mainly into the region of injury. They did not show specific association with initially necrotic tubules, but some of them located in close proximity to regenerating tubules. Nonlethally injured cells at the distal end of proximal tubules are likely to be the main source of tubular regeneration, and the transient appearance of interstitial myofibroblasts attached to the tubular basement membrane immediately after tubular necrosis might play a role in promoting cellular recovery in possible association with monocytes/macrophages in uranyl acetate-induced acute renal failure. PMID- 11021837 TI - A repertoire of cell cycle regulators whose expression is coordinated with human cytotrophoblast differentiation. AB - Although placental development depends on careful coordination of trophoblast proliferation and differentiation, little is known about the mitotic regulators that are key to synchronizing these events. We immunolocalized a broad range of these regulators in tissue sections of the maternal-fetal interface (first trimester through term) that contained floating villi (which include cytotrophoblasts differentiating into syncytiotrophoblasts) and anchoring villi (which include cytotrophoblasts differentiating into invasive cells). Trophoblast populations at the maternal-fetal interface stained for 16 of the cell cycle regulators whose expression we studied. The staining patterns changed as a function of both differentiation and gestational age. Differentiation along the invasive pathway was associated with entrance into, then permanent withdrawal from, the cell cycle, as evidenced by the orchestrated expression of cyclins, their catalytic subunits, and inhibitors. Surprisingly, we found coexpression of molecules that regulate different portions of the cell cycle in the syncytium. These data, which constitute one of the few examples to date of in situ localization of an extensive repertoire of mitotic regulators, provide the basis for studies aimed at understanding factors that lead to abnormal placentation. PMID- 11021838 TI - Glucose stimulation of transforming growth factor-beta bioactivity in mesangial cells is mediated by thrombospondin-1. AB - Glucose is a key factor in the development of diabetic complications, including diabetic nephropathy. The development of diabetic glomerulosclerosis is dependent on the fibrogenic growth factor, transforming growth factor-beta (TGF-beta). Previously we showed that thrombospondin-1 (TSP-1) activates latent TGF-beta both in vitro and in vivo. Activation occurs as the result of specific interactions of latent TGF-beta with TSP-1, which potentially alter the conformation of latent TGF-beta. As glucose also up-regulates TSP-1 expression, we hypothesized that the increased TGF-beta bioactivity observed in rat and human mesangial cells cultured with high glucose concentrations is the result of latent TGF-beta activation by autocrine TSP-1. Glucose-induced bioactivity of TGF-beta in mesangial cell cultures was reduced to basal levels by peptides from two different sequences that antagonize activation of latent TGF-beta by TSP, but not by the plasmin inhibitor, aprotinin. Furthermore, glucose-dependent stimulation of matrix protein synthesis was inhibited by these antagonist peptides. These studies demonstrate that glucose stimulation of TGF-beta activity and the resultant matrix protein synthesis are dependent on the action of autocrine TSP-1 to convert latent TGF-beta to its biologically active form. These data suggest that antagonists of TSP-dependent TGF-beta activation may be the basis of novel therapeutic approaches for ameliorating diabetic renal fibrosis. PMID- 11021839 TI - Quantitative ultrastructural analysis of a single spinal cord demyelinated lesion predicts total lesion load, axonal loss, and neurological dysfunction in a murine model of multiple sclerosis. AB - Infection of susceptible mice with Theiler's murine encephalomyelitis virus results in neurological dysfunction from progressive central nervous system demyelination that is pathologically similar to the human disease, multiple sclerosis. We hypothesized that the development of neuropathology proceeds down a final common pathway that can be accurately quantified within a single spinal cord lesion. To test this hypothesis, we conducted quantitative ultrastructural analyses of individual demyelinated spinal cord lesions from chronically infected mice to determine whether pathological variables assessed within a single lesion accurately predicted global assessments of morphological and functional disease course. Within lesions we assessed by electron microscopy the frequencies of normally myelinated, remyelinated, and demyelinated axons, as well as degenerating axons and intra-axonal mitochondria. The frequency of medium and large remyelinated fibers within a single lesion served as a powerful indicator of axonal preservation and correlated with preserved neurological function. The number of degenerating axons and increased intra-axonal mitochondria also correlated strongly with global measures of disease course, such as total lesion load, spinal cord atrophy, and neurological function. This is the first study to demonstrate that functional severity of disease course is evident within a single demyelinated lesion analyzed morphometrically at the ultrastructural level. PMID- 11021842 TI - Metastatic and nonmetastatic models of retinoblastoma. AB - To generate animal models of retinoblastoma that closely resemble metastatic and nonmetastatic human disease for the purposes of examining tumor biology and developing alternate treatments, human retinoblastoma cell lines were injected into the vitreal cavities of immunodeficient mice. Two reproducible animal models with contrasting biological behaviors analogous to human retinoblastoma have been developed. The Y79 retinoblastoma model demonstrated specific tumor evolution similar to that seen in human invasive and metastatic disease. Y79 retinoblastoma cells formed intraocular tumors that were initially confined to the vitreal cavity. Tumors progressively invaded the retina, subretinal space, choroid, optic nerve head, and anterior chamber of the eye. Tumors progressed into the subarachnoid space and focally invaded the brain. Metastases were detected in the contralateral optic nerve. Large tumors developed extraocular extensions. The histology of the tumors showed a poorly differentiated pattern with high mitotic rate, foci of necrosis, and calcification. The WERI-Rb model more closely resembled nonmetastatic human retinoblastoma. WERI- Rb tumors were localized in the eye with only anterior choroidal invasion at late stages. To examine potential biological differences in vitro, the retinoblastoma cell lines were cocultured with adherent choroid cells or adherent glioma cells which represent the targets of invasive retinoblastoma in vivo. Consistent with the in vivo observations, Y79 cells but not WERI-Rb cells adhere specifically to both the choroidal and the glioma cell lines. PMID- 11021840 TI - Novel human alpha9 acetylcholine receptor regulating keratinocyte adhesion is targeted by Pemphigus vulgaris autoimmunity. AB - Pemphigus vulgaris (PV) is a potentially fatal autoimmune mucocutaneous blistering disease. It was assumed that PV is caused by anti-desmoglein (Dsg) 3 autoimmunity because absorption of PV sera with a chimeric baculoprotein containing the Dsg 3 and IgG1 portions, rDsg3-Ig-His, eliminated disease-causing antibodies. In this study we demonstrate that rDsg3-Ig-His adsorbs out autoantibodies to different keratinocyte antigens, including a non-Dsg 3 130-kd polypeptide. Because the pool of disease-causing PV IgGs contains antibodies against the keratinocyte acetylcholine receptor (AChR), we sought to identify the targeted receptor(s). Preincubation of monkey esophagus with PV antibodies blocked specific staining of the keratinocyte cell membrane with rabbit monoepitopic antibody to alpha9 AChR, indicating that this first of its kind AChR with dual, muscarinic and nicotinic pharmacology is targeted by PV autoimmunity. Anti-alpha9 antibody stained keratinocytes in a fishnet-like intercellular pattern, and visualized a single band at approximately 50 kd in Western blots of keratinocyte membrane proteins. Using step-by-step reverse transcription polymerase chain reactions with primers based on known alpha9 sequence regions, we identified the complete reading frame of human alpha9. Its amino acid sequence showed 85% similarity with rat alpha9. Treatment of keratinocyte monolayers with anti-alpha9 antibody induced pemphigus-like acantholysis, which could be reversed either spontaneously or by using the cholinergic agonist carbachol. We conclude that alpha9 is coupled to physiological regulation of keratinocyte adhesion, and its interaction with PV IgG may lead to blister development. PMID- 11021843 TI - Quality of websites: kitemarking the west wind. PMID- 11021841 TI - p57(KIP2) is not mutated in hepatoblastoma but shows increased transcriptional activity in a comparative analysis of the three imprinted genes p57(KIP2), IGF2, and H19. AB - Hepatoblastomas (HBs), representing malignant liver tumors of childhood, show frequent loss of heterozygosity (LOH) in the chromosomal region 11p15.5. This loss is of maternal origin suggesting the presence of a monoallelically expressed tumor suppressor gene in this region. p57(KIP2) (KIP2) located at 11p15.5 is predominantly expressed from the maternal allele and encodes a cyclin-dependent kinase inhibitor. We screened a series of 56 HB tumors and five HB cell lines for allelic loss (LOH) of the KIP2 locus by microsatellite analysis and KIP2 coding sequence mutations by single-strand conformation polymorphism analysis. Although LOH at the KIP2 locus occurred in 25% of the cases, no mutations were found. Analysis of KIP2 mRNA expression by competitive reverse transcriptase-polymerase chain reaction revealed up-regulation in nine of 12 HBs compared to matching liver samples. In contrast, mRNA levels of the putative suppressor gene H19 on 11p15.5 were decreased in 10 of 12 tumors, indicating that KIP2 and H19 are not co-regulated in HBs. IGF2 mRNA expression was increased in 11 of 12 HB samples. All HBs showed monoallelic KIP2 expression. However, the overexpression of KIP2 in HBs with maternal loss of 11p15.5 suggests a reactivation of the paternal allele in these cases. Overexpression of KIP2 in HBs argues against a role as a HB suppressor gene. PMID- 11021844 TI - Fluoridation, fractures, and teeth. PMID- 11021845 TI - The risk of bias from omitted research. PMID- 11021846 TI - Electronics, clinicians, and the NHS. PMID- 11021847 TI - Cancer and insulin-like growth factor-I. A potential mechanism linking the environment with cancer risk. PMID- 11021848 TI - Cancer registries fear imminent collapse. PMID- 11021849 TI - UK death rates from breast cancer fall by a third. PMID- 11021850 TI - In brief PMID- 11021852 TI - Doctors doubt whether cancer targets can be met. PMID- 11021851 TI - UK drugs industry sets up trials register. PMID- 11021853 TI - Abortion pill is approved for sale in United States. PMID- 11021854 TI - Interferon treatment is beneficial in early multiple sclerosis PMID- 11021855 TI - Flu experts warn of need for pandemic plans. PMID- 11021856 TI - Netherlands bans cloning of human embryos for research. PMID- 11021857 TI - EU aims to reduce suicides. PMID- 11021858 TI - Twins' lawyers may demand change of venue for operation. PMID- 11021859 TI - More drugs pass the NICE test. PMID- 11021860 TI - The gene collector. PMID- 11021861 TI - Systematic review of water fluoridation. AB - OBJECTIVE: To review the safety and efficacy of fluoridation of drinking water. DESIGN: Search of 25 electronic databases and world wide web. Relevant journals hand searched; further information requested from authors. Inclusion criteria were a predefined hierarchy of evidence and objectives. Study validity was assessed with checklists. Two reviewers independently screened sources, extracted data, and assessed validity. MAIN OUTCOME MEASURES: Decayed, missing, and filled primary/permanent teeth. Proportion of children without caries. Measure of effect was the difference in change in prevalence of caries from baseline to final examination in fluoridated compared with control areas. For potential adverse effects, all outcomes reported were used. RESULTS: 214 studies were included. The quality of studies was low to moderate. Water fluoridation was associated with an increased proportion of children without caries and a reduction in the number of teeth affected by caries. The range (median) of mean differences in the proportion of children without caries was -5.0% to 64% (14.6%). The range (median) of mean change in decayed, missing, and filled primary/permanent teeth was 0.5 to 4.4 (2.25) teeth. A dose-dependent increase in dental fluorosis was found. At a fluoride level of 1 ppm an estimated 12.5% (95% confidence interval 7.0% to 21.5%) of exposed people would have fluorosis that they would find aesthetically concerning. CONCLUSIONS: The evidence of a beneficial reduction in caries should be considered together with the increased prevalence of dental fluorosis. There was no clear evidence of other potential adverse effects. PMID- 11021863 TI - Audit of oxygen prescribing before and after the introduction of a prescription chart. PMID- 11021862 TI - Community water fluoridation, bone mineral density, and fractures: prospective study of effects in older women. AB - OBJECTIVE: To determine whether fluoridation influences bone mineral density and fractures in older women. DESIGN: Multicentre prospective study on risk factors for osteoporosis and fractures. SETTING: Four community based centres in the United States. PARTICIPANTS: 9704 ambulatory women without bilateral hip replacements enrolled during 1986-8; 7129 provided information on exposure to fluoride. MAIN OUTCOME MEASURES: Bone mineral density of the lumbar spine, proximal femur, radius, and calcaneus plus incident fractures (fractures that occurred during the study) of vertebrae, hip, wrist, and humerus. RESULTS: Women were classified as exposed or not exposed or having unknown exposure to fluoride for each year from 1950 to 1994. Outcomes were compared in women with continuous exposure to fluoridated water for the past 20 years (n=3218) and women with no exposure during the past 20 years (n=2563). In women with continuous exposure mean bone mineral density was 2.6% higher at the femoral neck (0.017 g/cm(2), P<0.001), 2.5% higher at the lumbar spine (0.022 g/cm(2), P<0.001), and 1.9% lower at the distal radius (0.007 g/cm(2), P=0.002). In women with continuous exposure the multivariable adjusted risk of hip fracture was slightly reduced (risk ratio 0.69, 95% confidence interval 0.50 to 0.96, P=0.028) as was the risk of vertebral fracture (0.73, 0.55 to 0.97, P=0.033). There was a non-significant trend toward an increased risk of wrist fracture (1.32, 1.00 to 1.71, P=0.051) and no difference in risk of humerus fracture (0.85, 0.58 to 1.23, P=0.378). CONCLUSIONS: Long term exposure to fluoridated drinking water does not increase the risk of fracture. PMID- 11021864 TI - Refused and granted requests for euthanasia and assisted suicide in the Netherlands: interview study with structured questionnaire. PMID- 11021865 TI - "Old Man" (after marot) PMID- 11021866 TI - Do patients wish to be involved in decision making in the consultation? A cross sectional survey with video vignettes. AB - OBJECTIVE: To determine patients' preferences for a shared or directed style of consultation in the decision making part of the general practice consultation. DESIGN: Structured interview, with video vignettes of acted consultations. SETTING: 5 practices in Lothian, Scotland. PARTICIPANTS: 410 patients (adults and adults accompanying children) attending surgery appointments. MAIN OUTCOME MEASURES: Preference for shared or directed form of video vignette for five different presenting conditions. RESULTS: Patients varied in their preference for involvement in decision making in the consultation. Under multiple regression analysis, patients' preference was found to be independently predicted by the problem viewed (patients presented with physical problems preferred a directed approach), patients' age (patients aged 61 or older were more likely to prefer the directed approach), social class (social classes I and II were more likely to prefer the shared approach), and smoking status (smokers more likely to prefer the shared approach). Those patients who were able to answer (or who thought their doctor's style similar to those in the vignettes) were more likely to describe their own doctor's style as similar to their preferred style. No major association in preference was found with sex, frequency of attendance, or perceived chronic ill health. CONCLUSION: Patients may vary in their desire for involvement in decision making in consultations. Although this variation seems to depend on the presenting problem, age, social class, and smoking status, these associations are not absolute, with large minorities in each group. Doctors need the skills, knowledge of their patients, and the time to determine on which occasions, with which illnesses, and at which level their patients wish to be involved in decision making. PMID- 11021868 TI - How to die PMID- 11021867 TI - Clinical governance in primary care: knowledge and information for clinical governance. PMID- 11021869 TI - Back to basics on NHS networking. PMID- 11021870 TI - NHSnet in Scottish primary care: lessons for the future. AB - OBJECTIVE: To evaluate the primary care communications initiative, which introduced NHSnet to primary care in Scotland. DESIGN: Semi-structured telephone interviews, postal questionnaire. SETTING: All 15 Scottish health boards, random sample of 1 in 3 of all Scottish general practices. PARTICIPANTS: Information management and technology managers of health boards, 355 practice managers in the general practices. MAIN OUTCOME MEASURES: Variations between health boards in styles of project management, means of connection to NHSnet, costs to general practices, and training provided. Practices' levels of participation in initiative, initial use of NHSnet, and factors acting as incentives and disincentives to use of NHSnet. RESULTS: 99% of Scottish general practices agreed to participate in initiative. Health boards varied significantly in project management styles (from minimal to total control), the nature of the networks they established (intranets or direct connections), costs to practices (from nothing to pound125 per general practitioner per year), and training provided (from none to an extensive programme). In 56% of practices someone accessed NHSnet at least once a week. Practices varied considerably in amount of internet training received and staff groups targeted and in the intention to provide desktop access to NHSnet through a practice network. CONCLUSION: The initiative has successfully introduced a network that links Scottish general practices, health boards, and hospital trusts. However local variation in this "national" initiative may affect its use in primary care. Health authorities and general practices in England and Wales may wish to note these findings in order to avoid unhelpful variation. PMID- 11021872 TI - All in a day's work PMID- 11021871 TI - Recent advances: rheumatology. PMID- 11021873 TI - ABC of colorectal cancer. Molecular basis for risk factors. PMID- 11021874 TI - Data protection legislation: interpretation and barriers to research. PMID- 11021875 TI - Health care for asylum seekers. Main obstacles are inflexibility of NHS and bureaucracy of support systems. PMID- 11021876 TI - Health care for asylum seekers. General practitioners' knowledge of issues relating to asylum seekers is poor. PMID- 11021877 TI - Health care for asylum seekers. Germany has better method of dealing with asylum seekers' medical problems. PMID- 11021878 TI - Use of ambulatory blood pressure monitoring. Elegant new test needs clinical indication. PMID- 11021879 TI - The secret life of the NHS. Trust seems not to have understood function of walk in centre. PMID- 11021880 TI - The secret life of the NHS. Managers have invented new system for reducing waiting list times in Suffolk. PMID- 11021881 TI - Predicting left ventricular systolic dysfunction. Combining test results gives best predictive effect. PMID- 11021882 TI - Predicting left ventricular systolic dysfunction. Cut-off level was inappropriate. PMID- 11021883 TI - Randomised controlled trial of chest pain units is needed. PMID- 11021884 TI - Social cost-benefit analysis of teledermatology. Costs were understated. PMID- 11021885 TI - Life is as much a pain as it ever was. PMID- 11021886 TI - Paying for the NHS. Democratic control should not be dismissed. PMID- 11021887 TI - Paying for the NHS. What about health needs? PMID- 11021888 TI - Paying for the NHS. Spending should be decided by public and politicians. PMID- 11021889 TI - Paying for the NHS. People covered by private health insurance will not reduce consumption of NHS services. PMID- 11021890 TI - Minimising harm from hepatitis C virus needs better strategies. PMID- 11021891 TI - Combination treatment for hepatitis C is not being given. PMID- 11021892 TI - Out of Africa: some lessons for general practice/family medicine in developed countries? PMID- 11021893 TI - GP job satisfaction in 1987, 1990 and 1998: lessons for the future? AB - BACKGROUND: Job satisfaction is an important determinant of physician retention and turnover, and may also affect performance. Objective. Our aim was to investigate changes in GP job satisfaction from 1987 to 1998, covering a period of major change in the organization of British general practice. METHODS: Postal surveys of random national samples of GPs were carried out by separate groups of investigators in 1987, 1990 and 1998. In each survey, the questionnaire contained a standardized job satisfaction scale and a list of 14 job stressors. The final samples consisted of 1817 GPs in 1987 (response rate 45%), 917 GPs in 1990 (response rate 61%) and 1828 GPs in 1998 (response rate 49%). RESULTS: For both men and women, overall job satisfaction declined from 1987 to 1990 and then improved from 1990 to 1998, although satisfaction in 1998 remained below that in 1987. Women tended to report higher levels of satisfaction than men in all 3 years. Satisfaction with nine specific aspects of work showed dissimilar patterns of change over time. From 1987 to 1990, reported levels of stress increased for eight of 14 job stressors. Of these, three subsequently declined in 1998, two remained unchanged and three continued to increase. Of the six job stressors which showed no change from 1987 to 1990, five subsequently increased as sources of stress. Men and women differed in their sources of stress, but the differences were not consistent over time. CONCLUSIONS: The results suggest that GP job satisfaction has improved significantly from the low point reached following the introduction of the 1990/1991 NHS reforms, although reported levels of stress in relation to many aspects of work have continued to increase. The changes are discussed within the context of wider research into the determinants of GP job satisfaction in order to anticipate the likely effects on GPs of future organizational reforms. PMID- 11021894 TI - The General Practice Assessment Survey (GPAS): tests of data quality and measurement properties. AB - OBJECTIVES: The aim of this study was to describe the psychometric properties of the General Practice Assessment Survey (GPAS) and its acceptability to patients in the UK. GPAS comprises seven multiple item scales and two single item scales addressing nine key areas of primary care activity (access, technical care, communication, inter-personal care, trust, knowledge of patient, nursing care, receptionists and continuity of care). A further four single items relate to patients' perceptions of the GP's role in referral and co-ordination of care, their willingness to recommend their GP and their overall satisfaction with care received. METHODS: Two hundred consecutive patients attending routine consulting sessions at 55 inner London practices were invited to complete the GPAS questionnaire. The acceptability, reliability and validity of GPAS was assessed using standard psychometric techniques. RESULTS: Out of 11 000 patients, 7247 (66%) completed a questionnaire in a GP surgery. Fifty-five out of a separate sample of 77 patients attending one practice completed a second questionnaire mailed to them 1 week following their attendance. GPAS was acceptable to patients as evidenced by low proportions of missing data for all items, and a full range of possible scores for all but one of the nine scales. Reliability of the instrument was good. Multiple item scales had excellent internal consistency, high item-total correlations, and test-retest reliability. Scaling assumptions were confirmed, with six of the seven scales achieving 100% scaling success (convergent and discriminant validity). Construct validity was evident, although this requires further evaluation against external measures. CONCLUSIONS: GPAS is a useful instrument for assessing several important dimensions of primary care. It is acceptable, reliable and valid, and has the potential for versatility in mode of administration. It will be a useful instrument for practices, primary care groups and primary care researchers evaluating key areas of primary care activity. Further work is required to evaluate its performance in non-inner-city settings and to evaluate further its validity against external criteria. PMID- 11021895 TI - Antibiotics for coughing in general practice: a qualitative decision analysis. AB - BACKGROUND: In family practice, medical decisions are prompted most often by complaints about coughing. There is no single yardstick for the differential diagnosis of respiratory tract infections (RTIs). In 80% of cases, the excessive use of antibiotics in the treatment of RTIs is caused by the prescription behaviour of GPs. OBJECTIVE: Our aim was to explicate GPs' diagnostic (and therapeutic) decisions regarding adult patients who consult them with complaints about coughing, and to investigate what determines decision making. METHODS: Exploratory, descriptive focus groups were held with GPs. Hypotheses were generated on the basis of 'qualitative content analysis'. Results. Twenty-four GPs participated in four semi-structured group discussions. In order to differentiate RTIs from other possible diagnoses, less likely diagnoses were not ruled out explicitly. In the case of suspected RTI, there was a low degree of certainty in the differentiation between RTIs (e.g. between bronchitis and pneumonia). Clinical signs and symptoms, which determine the probability of disease, often left GPs with reasonable diagnostic doubt. In the end, the decision whether or not to prescribe antibiotics was taken. GPs' prescription behaviour was also determined by doctor- and patient-related factors (e.g. having missed pneumonia once, patient expectations). The 'chagrin factor' explains why these factors lead to a shift in the action threshold, in favour of antibiotics. CONCLUSION: This inductive research method enabled the generation of meaningful hypotheses regarding the complex decision processes pursued by GPs. The authors are developing an educational intervention that builds on these findings, focusing on the prescribing decision. PMID- 11021896 TI - Predictors of an antibiotic prescription by GPs for respiratory tract infections: a pilot. AB - BACKGROUND: Antibiotics are over-prescribed for respiratory tract infections in Australia. OBJECTIVES: The aim of this study was to describe the clinical predictors of GPs' prescribing of antibiotics. METHODS: We used Clinical Judgment Analysis to study the responses of GPs to hypothetical paper-based vignettes of a 20-year-old with a respiratory tract infection. The nature of four symptoms and signs (colour of nasal mucous discharge; soreness of the throat; presence of fever; and whether any cough was productive of sputum) was varied and their effect on prescribing measured using logistic regression. RESULTS: Twenty GPs participated. The nature of each symptom and sign significantly predicted prescribing of an antibiotic. Cough productive of yellow sputum; presence of sore throat; fever; and coloured nasal mucus increased the probability of an antibiotic being prescribed. CONCLUSIONS: GPs are influenced by clinical signs and symptoms to use antibiotics for respiratory infections for which there is poor evidence of efficacy from the literature. PMID- 11021897 TI - Observational study of home visits in Slovene general practice: patient characteristics, practice characteristics and health care utilization. AB - BACKGROUND: Home visits are an important way of delivering primary health care, but there is a long-term decrease in home visit rates in many countries. OBJECTIVE: The aim of this study was to evaluate patient characteristics, morbidity, functional status, quality of life, satisfaction with care, practice characteristics and health care utilization in general practice patients visited at home at least once in a study year. METHODS: The design of the study was a cross-sectional survey of the patients of a stratified sample of 36 GP offices in Slovenia using a self-administered questionnaire. Sixty consecutive patients in sampled practices contacting the doctor in the office in the study period in March 1998 were included in the analysis. The age, sex, educational status, residence, presence of chronic condition, measures of anxiety or depressive symptoms, rates of patients who expressed a need for emergency care in 1 year, rates of self-care, measures of functional status, quality of life, satisfaction with care, rates of using GP practice visits and out-of-hours services and rates of using specialist or hospital services were recorded in a home-visited group versus a non-visited group. RESULTS: A total of 277 patients (15.4%) were reported to have at least one visit in the study year. Patients visited in their homes were older, predominantly female, better educated, had lower perceptions of their functional status and well-being and they used primary health services more frequently than others. Their GPs were more likely to be males, and were more likely to practise in rural areas, in solo practices as private practitioners. CONCLUSION: Home visits remain an important part of GP work in countries in transition, such as Slovenia, especially for more seriously ill patients. PMID- 11021898 TI - Time changes in new cases of ischaemic heart disease in general practice. AB - BACKGROUND: There are no recent studies of the presentation of ischaemic heart disease (IHD) in general practice. What information exists is derived from the secondary care setting, where seasonal and daily variation has been reported in admissions for IHD. There are epidemiological studies that show a falling incidence and mortality for IHD. It is not clear, however, if this is also the case in clinical general practice. OBJECTIVES: The aims of the present study were to (i) estimate the number of cases of IHD in general practice populations; (ii) determine the recorded diagnosis and time of first presentation of IHD during a 5 year period; and (iii) perform time series analysis on the above data. METHODS: The design of the study was a retrospective survey, using MIQUEST software, of computer databases in five general practices with a combined population of nearly 40,000 patients. The five practices were selected randomly from volunteering practices in the Trent Focus Collaborative Research Network. All patients with a new diagnosis of IHD recorded between January 1993 and December 1997 inclusive were included in the study. The number of new cases of IHD, the recorded diagnosis and time of first presentation of IHD were the main outcome measures. RESULTS: A total of 644 new cases of IHD were identified during the study period: 54.0% 'angina pectoris', 26. 9% 'acute myocardial infarction', 18.8% 'ischaemic heart disease', 0. 3% 'coronary atherosclerosis'. Time series analysis reveals a seasonal and weekly pattern to new cases of IHD, with peak cases occurring in January and on Mondays/Fridays. A downward trend was detected for new cases of IHD (all diagnoses) over the 5 year period, and for new cases of IHD (excluding acute myocardial infarction). An upward trend was observed for new cases of acute myocardial infarction. CONCLUSION: Presentation of IHD in general practice varies according to season and day of the week. The proportion of new cases recorded as 'angina pectoris' or 'ischaemic heart disease' is falling, and this decline masks a rise in the incidence recordings of 'acute myocardial infarction'. PMID- 11021899 TI - High dose proton pump inhibitor response as an initial strategy for a clinical diagnosis of gastro-oesophageal reflux disease (GERD). Swedish multi-centre group in primary health care. AB - BACKGROUND: The diagnosis of gastro-oesophageal reflux disease (GERD) in primary care rests primarily on symptoms. Oesophageal acid exposure is the most important pathogenic factor and it is likely that symptom response to acid inhibition also identifies patients with GERD. OBJECTIVE: The aim of this study was to evaluate the outcome of a symptom-based strategy in the management of GERD patients in primary care. METHODS: Patients in general practice with main symptoms of at least moderate heartburn or regurgitation were given omeprazole 20 mg b.i.d. openly for 7 days (first phase). Responders with later relapse were randomized to double-blind treatment with omeprazole 20 mg o.m. or placebo for 2 weeks (second phase). A response in both phases was defined as a decrease by at least three grades on a seven-grade Likert scale and no more than mild intensity of the main symptom. RESULTS: Of the 362/371 recruited patients who were evaluated in the first phase, 73% were responders. A total of 174/179 patients with a relapse were assessed in the second phase, and 74 and 28% in the omeprazole and placebo group, respectively, were defined as responders (P: < 0.001, 95% confidence interval 33 59). CONCLUSIONS: GERD patients are highly responsive to omeprazole 20 mg b.i.d. They are equally responsive to omeprazole 20 mg o.m. at symptomatic relapse, but have a low response rate to placebo. Omeprazole is a valuable therapeutic instrument to detect and treat patients with GERD in general practice. PMID- 11021900 TI - Health problems in people with intellectual disability in general practice: a comparative study. AB - In a GP database, 318 people with intellectual disability (ID) appeared to have 2.5 times more health problems than people without ID. This short report deals with the nature of the health problems. Consequences for health care policy are discussed. PMID- 11021901 TI - Factors associated with fatigue in a family medicine clinic in the United Arab Emirates. AB - BACKGROUND: Fatigue is a common symptom in Family Medicine and it has many associated factors. The Arabian Gulf provides a unique setting for studying these factors, in particular the UAE where rapid development has been a prominent feature. OBJECTIVES: The aim of the study was to sample a group of GP attenders and examine the factors which were associated with fatigue in the UAE. METHODS: A fatigue scale, psychological questionnaire, detailed history, physical examination and laboratory testing were administered to a sample of attenders at a Family Medicine clinic. RESULTS: Fatigue was more prevalent than in western studies (males 34.0%, females 38. 2%). It was strongly associated with anxiety, especially in younger adults, and it has been recognized that rapid social change is felt most acutely in young adults and adolescents. Depression in females was also a factor. Lack of exercise, obesity and illiteracy played a minor role in the severity of fatigue. CONCLUSIONS: Fatigue appears to be a cultural 'idiom of distress', a way of expressing anxiety or depression in a rapidly changing society. PMID- 11021902 TI - Differences in physician utilization between Aboriginal and non-Aboriginal children. AB - OBJECTIVE: In the present study, we examined the factors affecting Aboriginal children's visits to a medical practitioner and compared them with non-Aboriginal children. METHODS: We selected five Aboriginal communities and four neighbouring non-Aboriginal communities, and conducted a door-to-door survey, covering all children born after 1983. Of an initial sample of 1013 children, 896 (response rate 89.92% for non-Aboriginal children and 85.87% for Aboriginal children) completed the questionnaire for analysis. RESULTS: In all, 896 children of non mixed lineage with an age range of 0-12 years were collected into the study, including 316 Aborigines and 580 non-Aborigines. A higher percentage of non Aboriginal children had more national health insurance coverage than Aboriginal children. The ratio of parents using the services of an out of community medical practitioner when their children were sick was higher for Aboriginal parents than for non-Aborigines. Medical injection frequency was higher in Aboriginal children. Linear regression was used to examine the factors affecting the frequency of physician utilization in the preceding month. CONCLUSION: A lower national health insurance coverage rate, and a higher rate of intramuscular injections for Aboriginal children plus difficulties in access to medical resources due to travel time and travel distance are still major problems for the Aborigines. PMID- 11021903 TI - Using video-recorded consultations for research in primary care: advantages and limitations. AB - BACKGROUND: Video-recording primary care consultations is an established technique for primary care research. Despite the widespread use of video recording to help answer a variety of research questions, little is known about how this recording technique influences the findings of studies in which it is employed. OBJECTIVE: This article investigates how video-recorded consultations have been used in research and discusses how this technique may influence both the internal and external validity of studies. CONCLUSION: Using video-recorded consultations for research purposes may cause bias in the characteristics of doctors and patients who agree to participate in research. There is little evidence, however, that video-recording influences the behaviour of either GPs or patients. Recommendations are made for researchers who are considering using video-recorded consultations in their research. PMID- 11021904 TI - A preliminary study of the decision-making process within general practice. AB - OBJECTIVE: The aim of the present study was to explore the factors that contribute to the process of decision making within general practice, over and above evidence-based information. METHODS: A qualitative study was conducted using semi-structured interviews on a purposeful sample of GPs, based in the South West of England. Each interview was tape-recorded and transcribed verbatim. RESULTS: Five broad categories emerged from the data: practitioner; patient; practitioner-patient relationship; verbal and non-verbal communication; evidence based medicine; and external factors. CONCLUSION: The nature of general practice is such that the process of making clinical decisions is complex. In an era when GPs are being overwhelmed by evidence-based information, consideration needs to be given to the implications that the nature of the decision-making process has upon the way 'evidence' is constructed and promoted within general practice. PMID- 11021905 TI - Reflexivity--a strategy for a patient-centred approach in general practice. AB - Reflexivity as a strategy in general practice can be used to implement a patient centred approach in the consultation. General practice has long represented a tradition attempting to integrate both illness and disease. For the GP, it is natural to focus on the patient's whole situation, and the GP's experience with patients is often based on a long-term relationship. Reflexivity implies having a self-conscious account of the production of knowledge as it is being produced. We believe that GPs can gain access to additional knowledge by consciously using reflexivity as a strategy in the consultation. In the present article, we discuss reflexivity in relation to the notions of empathy, personal experience and self knowledge. By using reflexivity in order to rely on personal experience, the GP can gain access to patients' understanding of their health. Reflexivity can be a valuable concept for the GP in patient-centred medicine and can contribute to bridging the gap between the patient's perspective and the doctor's understanding of the patient's health. PMID- 11021906 TI - Selections from current literature. Recent advances in conjugated pneumococcal vaccination. AB - This article reviews the current literature and recent updates with regard to childhood vaccination for Streptococcus pneumonia including: studies in immunology with antibody titres, dosages of conjugated vaccines, carriage rates of Streptococcus, side effects, comparison in certain disease states and comparison between vaccines. PMID- 11021907 TI - Evidence-based management of groin hernia in primary care--a systematic review. AB - BACKGROUND: National clinical guidelines on the surgical management of groin hernia have been published by the Royal College of Surgeons of England. There is also a need for guidance on the management of pre- and post-hernia repair patients in primary care, in areas such as diagnosis, referral and advice on recuperation. OBJECTIVE: The purpose of the present study was to determine best practice in primary care aspects of managing groin hernia in adults, by examination of the evidence base. METHOD: A systematic review of the available evidence was carried out, searching the major electronic databases: Medline, the Cochrane Library, Embase, Assia, Helmis, Cinahl and Psyclit. Key search terms were hern$, inguinal, femoral, groin, truss$, with searches limited to human adult subjects and the English language. RESULTS: Robust research on groin hernia is concerned almost exclusively with the in-patient surgical management of patients undergoing primary elective hernia repair. The areas with which this review was concerned, principally diagnosis, referral and advice about return to work, are areas in which it is more difficult to conduct robustly designed studies. Perhaps because of this, the evidence base on the non-surgical aspects of management is of poor methodological quality, being based primarily on expert opinion, reviews of clinical practice and experience, surveys, descriptive case studies and clinical audits. CONCLUSIONS: As the research in this area is generally of poor quality, strong conclusions are precluded, but it is possible to define best practice in some areas of care. In relation to diagnosis, GPs should distinguish correctly between a femoral and inguinal hernia because of the increased risks of strangulation and incarceration associated with the former. Due to clinical inaccuracy, the identification of whether a hernia is direct or indirect is not a good basis on which to base decision making regarding referral for elective repair. The risks associated with surgical repair are those of the normal range found for any procedure. Decisions about the fitness of patients for surgery in this instance are not procedure specific, and therefore the decisions about elective repair especially in older patients should be considered in terms of quality of life and patient choice rather than increased risks with surgical repair. Further research is required to address the gap in the evidence for the management of groin hernia within the primary care sector. PMID- 11021909 TI - WONCA news : An international forum for family doctors PMID- 11021908 TI - ICPC-2-E: the electronic version ICPC-2. PMID- 11021910 TI - Functional genomic studies of dihydroxyacetone utilization in Escherichia coli. PMID- 11021911 TI - The causes of Pseudomonas diversity. PMID- 11021912 TI - An interactive web-based Pseudomonas aeruginosa genome database: discovery of new genes, pathways and structures. AB - Using the complete genome sequence of Pseudomonas: aeruginosa PAO1, sequenced by the Pseudomonas: Genome Project (ftp://ftp.pseudomonas. com/data/pacontigs.121599), a genome database (http://pseudomonas. bit.uq.edu.au/) has been developed containing information on more than 95% of all ORFs in Pseudomonas: aeruginosa. The database is searchable by a variety of means, including gene name, position, keyword, sequence similarity and Pfam domain. Automated and manual annotation, nucleotide and peptide sequences, Pfam and SMART domains (where available), Medline and GenBank links and a scrollable, graphical representation of the surrounding genomic landscape are available for each ORF. Using the database has revealed, among other things, that P. aeruginosa contains four chemotaxis systems, two novel general secretion pathways, at least three loci encoding F17-like thin fimbriae, six novel filamentous haemagglutinin like genes, a number of unusual composite genetic loci related to vgr/RHS: elements in Escherichia coli, a number of fix-like genes encoding a micro-oxic respiration system, novel biosynthetic pathways and 38 genes containing domains of unknown function (DUF1/DUF2). It is anticipated that this database will be a useful bioinformatic tool for the Pseudomonas: community that will continue to evolve. PMID- 11021913 TI - Monitoring genome evolution ex vivo: reversible chromosomal integration of a 106 kb plasmid at two tRNA(Lys) gene loci in sequential Pseudomonas aeruginosa airway isolates. AB - The genome rearrangements in sequential Pseudomonas aeruginosa clone K isolates from the airways of a patient with cystic fibrosis were determined by an integrated approach of mapping, sequencing and bioinformatics. Restriction mapping uncovered an 8.9 kb deletion of PAO sequence between phnAB and oprL in clone K, and two 106 kb insertions either adjacent to this deletion or several hundred kilobases away, close to the pilA locus. These 106 kb blocks of extra DNA also co-existed as the circular plasmid pKLK106 in several clone K isolates and were found to be closely related to plasmid pKLC102 in P. aeruginosa clone C isolates. The breakpoints of the deletion in clone K and the attB-attP sequences for the reversible integration of the plasmid in clones C and K were located within the 3' end of the lysine tRNA structural genes (att site). pKLK106 sequentially recombined with either of the two tRNA(Lys) genes in clone K isolates. The att site of the pilA hypervariable region has been utilized by clone C to target its plasmid pKLC102 into the chromosome; the att site of the phnAB-oprL region has been employed by strain PAO to incorporate a DNA block encoding pyocin, transposases and IS elements. The use of typical phage attachment sites by conjugative genetic elements could be one of the major mechanisms used by P. aeruginosa to generate the mosaic genome structure of blocks of species-, clone- and strain-specific DNA. The example described here demonstrates the potential impact of systematic genome analysis of sequential isolates from the same habitat on our understanding of the evolution of microbial genomes. PMID- 11021914 TI - Phylogeny of the replication regions of pPT23A-like plasmids from Pseudomonas syringae. AB - It was previously shown that most Pseudomonas syringae strains contain one or more plasmids with cross-hybridizing replication regions and other areas of homology, and these plasmids were designated the pPT23A-like family. The majority of these plasmids encode genes conferring epiphytic fitness or resistance to antibacterial compounds and those investigated in this study are essential for pathogenicity or increased virulence. The phylogeny of 14 pPT23A-like plasmids from five P. syringae pathovars was studied by comparing a fragment of the sequence of their repA genes (encoding a replicase essential for replication). In the phylogenetic tree obtained, four groups (< or =88.8% identity between their members) could be identified. The first group contained the plasmids from three P. syringae pv. tomato strains, a P. syringae pv. apii strain and five out of the seven P. syringae pv. syringae strains, with identity ranging between 88.8 and 100%. The clustering of the pv. syringae strains did not reflect host specialization or previously reported phylogenetic relationships. The second group contained the plasmids from two strains of pv. glycinea and pv. tomato (95.5% identity), and it also included the previously sequenced replicon of a pathogenicity plasmid from P. syringae pv. phaseolicola. The plasmids from the remaining two pv. syringae strains were distantly related to the other plasmid sequences. Hybridization experiments using different genes or transposable elements previously described as plasmid-borne in P. syringae, showed that the gene content of highly related plasmids could be dissimilar, suggesting the occurrence of major plasmid reorganizations. Additionally, the phylogeny of the different native plasmids did not always correlate with the phylogeny of their harbouring strains, as determined by the analysis of extragenic repetitive consensus (ERIC) and arbitrarily primed PCR (AP-PCR) products. Collectively, these results suggest that pPT23A-like plasmids were, in most cases, acquired early during evolution. PMID- 11021915 TI - Phylogeny of the genus Pseudomonas: intrageneric structure reconstructed from the nucleotide sequences of gyrB and rpoD genes. AB - Phylogenetic analysis of the genus Pseudomonas: was conducted by using the combined gyrB and rpoD nucleotide sequences of 31 validly described species of Pseudomonas: (a total of 125 strains). Pseudomonas: strains diverged into two major clusters designated intrageneric cluster I (IGC I) and intrageneric cluster II (IGC II). IGC I was further split into two subclusters, the 'P: aeruginosa complex', which included P: aeruginosa, P: alcaligenes, P: citronellolis, P: mendocina, P: oleovorans and P: pseudoalcaligenes, and the 'P: stutzeri complex', which included P: balearica and P: stutzeri. IGC II was further split into three subclusters that were designated the 'P: putida complex', the 'P: syringae complex' and the 'P: fluorescens complex'. The 'P: putida complex' included P: putida and P: fulva. The 'P: syringae complex' was the cluster of phytopathogens including P: amygdali, P: caricapapayae, P: cichorii, P: ficuserectae, P: viridiflava and the pathovars of P. savastanoi and P. syringae. The 'P. fluorescens complex' was further divided into two subpopulations, the 'P. fluorescens lineage' and the 'P. chlororaphis lineage'. The 'P. fluorescens lineage' contained P. fluorescens biotypes A, B and C, P. azotoformans, P. marginalis pathovars, P. mucidolens, P. synxantha and P. tolaasii, while the 'P. chlororaphis lineage' included P. chlororaphis, P. agarici, P. asplenii, P. corrugata, P. fluorescens biotypes B and G and P. putida biovar B. The strains of P. fluorescens biotypes formed a polyphyletic group within the 'P. fluorescens complex'. PMID- 11021916 TI - Quantification of biofilm structures by the novel computer program COMSTAT. AB - The structural organization of four microbial communities was analysed by a novel computer program, COMSTAT, which comprises ten features for quantifying three dimensional biofilm image stacks. Monospecies biofilms of each of the four bacteria, Pseudomonas: putida, P. aureofaciens, P. fluorescens and P. aeruginosa, tagged with the green fluorescent protein (GFP) were grown in flow chambers with a defined minimal medium as substrate. Analysis by the COMSTAT program of four variables describing biofilm structure - mean thickness, roughness, substratum coverage and surface to volume ratio - showed that the four Pseudomonas: strains represent different modes of biofilm growth. P. putida had a unique developmental pattern starting with single cells on the substratum growing into micro-colonies, which were eventually succeeded by long filaments and elongated cell clusters. P. aeruginosa colonized the entire substratum, and formed flat, uniform biofilms. P. aureofaciens resembled P. aeruginosa, but had a stronger tendency to form micro colonies. Finally, the biofilm structures of P. fluorescens had a phenotype intermediate between those of P. putida and P. aureofaciens. Analysis of biofilms of P. aureofaciens growing on 0.03 mM, 0.1 mM or 0.5 mM citrate minimal media showed that mean biofilm thickness increased with increasing citrate concentration. Moreover, biofilm roughness increased with lower citrate concentrations, whereas surface to volume ratio increased with higher citrate concentrations. PMID- 11021917 TI - Experimental reproducibility in flow-chamber biofilms. AB - The structural organization of microbial communities is influenced by many factors, e.g. nutrient composition, shear stress and temperature. This paper presents a general method for quantitative comparison of biofilm structures and assessment of experimental reproducibility between independent biofilm experiments. By using a novel computer program, COMSTAT, biofilm structures of Pseudomonas aeruginosa and an isogenic rpoS mutant were quantified. The strains were tagged with the green fluorescent protein (GFP) and grown in flow chambers with a defined minimal medium as substrate. Three independent rounds of biofilm experiments were performed and in each round, each of the two variants was grown in two separate channels. Nine image stacks were acquired in each channel 146 h after inoculation. An analysis of variance model incorporating the factors experiment round, bacterial strain, channel number and image stack number was used to analyse the data calculated by COMSTAT. Experimental reproducibility was verified by estimating the magnitude of the variance of the effects round (sigma(2)R) and the interaction between bacterial strain and round (sigma(2)BR). Mean thickness of the wild-type and rpoS mutant biofilms was estimated at 6.31 microm (SE 0.81 microm) and 16.85 microm (SE 0.87 microm), respectively. PMID- 11021918 TI - Iron regulation of the hcnABC genes encoding hydrogen cyanide synthase depends on the anaerobic regulator ANR rather than on the global activator GacA in Pseudomonas fluorescens CHA0. AB - Pseudomonas fluorescens CHA0 produces hydrogen cyanide (HCN), a secondary metabolite that substantially contributes to this strain's biocontrol ability. Cyanogenesis is induced by oxygen-limiting conditions, but abolished by iron depletion. In P. fluorescens, the anaerobic regulator ANR and the global activator GacA are both required for the maximal expression of the HCN biosynthetic genes hcnABC. The molecular basis of this regulation by ANR and GacA was investigated under conditions of oxygen and iron limitation. A promoter deletion analysis using a translational hcnA'-'lacZ fusion revealed that a conserved FNR/ANR recognition sequence in the -40 promoter region was necessary and sufficient for the regulation by ANR in response to oxygen limitation. Stimulation of hcnA'-'lacZ expression by the addition of iron also depended on the presence of ANR and the FNR/ANR box, but not on GacA, suggesting that in addition to acting as an oxygen-sensitive protein, ANR also responds to iron availability. Expression of the translational hcnA'-'lacZ fusion remained GacA dependent in hcn promoter mutants that were no longer responsive to ANR, in agreement with earlier evidence for a post-transcriptional regulatory mechanism under GacA control. These data support a model in which cyanogenesis is sequentially activated by ANR at the level of transcription and by components of the GacA network at the level of translation. PMID- 11021919 TI - Vanadium interferes with siderophore-mediated iron uptake in Pseudomonas aeruginosa. AB - Vanadium is a metal that under physiological conditions can exist in two oxidation states, V(IV) (vanadyl ion) and V(V) (vanadate ion). Here, it was demonstrated that both ions can form complexes with siderophores. Pseudomonas aeruginosa produces two siderophores under iron-limiting conditions, pyoverdine (PVD) and pyochelin (PCH). Vanadyl sulfate, at a concentration of 1-2 mM, strongly inhibited growth of P. aeruginosa PAO1, especially under conditions of severe iron limitation imposed by the presence of non-utilizable Fe(III) chelators. PVD-deficient mutants were more sensitive to vanadium than the wild type, but addition of PVD did not stimulate their growth. Conversely, PCH negative mutants were more resistant to vanadium than the wild-type strain. Both siderophores could bind and form complexes with vanadium after incubation with vanadyl sulfate (1:1, in the case of PVD; 2:1, in the case of PCH). Although only one complex with PVD, V(IV)-PVD, was found, both V(IV)- and V(V)-PCH were detected. V-PCH, but not V-PVD, caused strong growth reduction, resulting in a prolonged lag phase. Exposure of PAO1 cells to vanadium induced resistance to the superoxide-generating compound paraquat, and conversely, exposure to paraquat increased resistance to V(IV). Superoxide dismutase (SOD) activity of cells grown in the presence of V(IV) was augmented by a factor of two. Mutants deficient in the production of Fe-SOD (SodB) were particularly sensitive to vanadium, whilst sodA mutants deficient for Mn-SOD were only marginally affected. In conclusion, it is suggested that V-PCH catalyses a Fenton-type reaction whereby the toxic superoxide anion O(2)- is generated, and that vanadium compromises PVD utilization. PMID- 11021920 TI - Heterogeneity of iron bioavailability on plants assessed with a whole-cell GFP based bacterial biosensor. AB - Ferric iron is an essential element for microbial growth but its water solubility in aerobic environments is considered to be low. Thus it is a limiting resource for which microbes must compete in natural habitats. Since competition for iron occurs at the level of individual cells, knowledge of the variability in iron bioavailability to such individuals is required to assess the nature of the competition in these habitats. Ferric iron availability to cells of Pseudomonas syringae was assessed by quantifying the fluorescence intensity of single cells harbouring a plasmid-borne transcriptional fusion of an iron-regulated promoter from a locus encoding a membrane receptor for a pyoverdine siderophore with a reporter gene encoding green fluorescent protein (GFP) following fluorescence microscopy. Cells of this iron biosensor exhibited iron-dependent GFP fluorescence that was inversely proportional to the amount of iron added to the media, and which differed by over 20-fold in iron-replete compared to iron deplete culture media. Cells cultured in a medium of a given iron content exhibited a very narrow range of fluorescence intensities. In contrast, the fluorescence intensity of cells of the biosensor strain recovered from the rhizosphere or phylloplane of inoculated bean plants varied greatly. The distribution of fluorescence intensities was strongly right-hand skewed, with about 10% of the cells exhibiting substantially higher GFP fluorescence than that of the median cell. Cells of a positive control strain, harbouring a fusion of the constitutive nptII promoter with the gfp reporter gene, exhibited uniform GFP fluorescence both in culture media and on plants. These results indicate that there is substantial heterogeneity of iron biovailability to cells of P. syringae on plants, with only a small subset of cells experiencing low iron availability. Such heterogeneity places constraints on models of interactions of bacteria in natural habitats that are based on competition for limited iron. PMID- 11021921 TI - Regulatory interactions between the Hrp type III protein secretion system and coronatine biosynthesis in Pseudomonas syringae pv. tomato DC3000. AB - In P. syringae, the co-ordinated regulation of different systems required for pathogenicity and virulence seems logical but has not been established. This question was addressed in the present study by analysing production of the phytotoxin coronatine (COR) in defined hrp/hrc mutants of P. syringae pv. tomato DC3000. COR was produced in vitro by mutants of DC3000 defective in hrcC, which encodes an outer-membrane protein required for type III-mediated secretion. When inoculated in plants, hrcC mutants produced chlorotic regions indicative of COR production, but lacked the necrotic lesions produced by the wild-type DC3000. Furthermore, a DC3000 mutant containing a polar mutation in hrcC, which inactivates hrcC, hrpT and hrpV, produced significantly higher amounts of COR than the wild-type strain in vitro. This mutant was able to produce COR earlier and at lower cell densities than the wild-type. The results indicate that the hrp/hrc secretion system is not required for COR production, but mutations in this system may have regulatory effects on the production of virulence factors such as COR. PMID- 11021922 TI - Temperature-responsive genetic loci in the plant pathogen Pseudomonas syringae pv. glycinea. AB - Plant-pathogenic bacteria may sense variations in environmental factors, such as temperature, to adapt to plant-associated habitats during pathogenesis or epiphytic growth. The bacterial blight pathogen of soybean, Pseudomonas syringae pv. glycinea PG4180, preferentially produces the phytotoxin coronatine at 18 degrees C and infects the host plant under conditions of low temperature and high humidity. A miniTn5-based promoterless glucuronidase (uidA) reporter gene was used to identify genetic loci of PG4180 preferentially expressed at 18 or 28 degrees C. Out of 7500 transposon mutants, 61 showed thermoregulated uidA expression as determined by a three-step screening procedure. Two-thirds of these mutants showed an increased reporter gene expression at 18 degrees C whilst the remainder exhibited higher uidA expression at 28 degrees C. MiniTn5-uidA insertion loci from these mutants were subcloned and their nucleotide sequences were determined. Several of the mutants induced at 18 degrees C contained the miniTn5-uidA insertion within the 32.8 kb coronatine biosynthetic gene cluster. Among the other mutants with increased uidA expression at 18 degrees C, insertions were found in genes encoding formaldehyde dehydrogenase, short-chain dehydrogenase and mannuronan C-5-epimerase, in a plasmid-borne replication protein, and in the hrpT locus, involved in pathogenicity of P. syringae. Among the mutants induced at 28 degrees C, insertions disrupted loci with similarities to a repressor of conjugal plasmid transfer, UV resistance determinants, an isoflavanoid-degrading enzyme, a HU-like DNA-binding protein, two additional regulatory proteins, a homologue of bacterial adhesins, transport proteins, LPS synthesis enzymes and two proteases. Genetic loci from 13 mutants did not show significant similarities to any database entries. Results of plant inoculations showed that three of the mutants tested were inhibited in symptom development and in planta multiplication rates. Temperature-shift experiments suggested that all of the identified loci showed a rather slow induction of expression upon change of temperature. PMID- 11021923 TI - The biocontrol strain Pseudomonas fluorescens F113 produces the Rhizobium small bacteriocin, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone, via HdtS, a putative novel N-acylhomoserine lactone synthase. AB - Several different species of Pseudomonas: produce N:-acylhomoserine lactones (AHLs), quorum-sensing signal molecules which are involved in the cell-density dependent control of secondary metabolite and virulence gene expression. When Pseudomonas fluorescens F113 was cross-streaked against AHL biosensors capable of sensitively detecting either short (C(4)-C(8)) or long (C(10)-C(14)) acyl chain AHLs, no activity was detectable. However, by extracting cell-free stationary phase culture supernatants with dichloromethane followed by reverse-phase HPLC, three distinct fractions were obtained capable of activating the AHL biosensors. Three AHLs were subsequently characterized using high-resolution MS and chemical synthesis. These were (i) N:-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone (3OH, C(14:1)-HSL), a molecule previously known as the Rhizobium leguminosarum small bacteriocin as a consequence of its growth inhibitory properties, (ii) N: decanoylhomoserine lactone (C(10)-HSL) and (iii) N:-hexanoylhomoserine lactone (C(6)-HSL). A gene (hdtS) capable of directing synthesis of all three P. fluorescens AHLs in Escherichia coli was cloned and sequenced. In vitro transcription/translation of hdtS yielded a protein of approximately 33 kDa capable of directing the synthesis of 3OH, C(14:1)-HSL, C(10)-HSL and C(6)-HSL in E. coli. HdtS does not belong to either of the known AHL synthase families (LuxI or LuxM) and is related to the lysophosphatidic acid acyltransferase family. HdtS may therefore constitute a member of a third protein family capable of AHL biosynthesis. PMID- 11021924 TI - Detection of N-acylhomoserine lactones in lung tissues of mice infected with Pseudomonas aeruginosa. AB - The pathogenesis of Pseudomonas aeruginosa is associated with expression of virulence factors, many of which are controlled by two N:-acylhomoserine lactone (AHL)-based quorum-sensing systems. Escherichia coli strains equipped with a luxR based monitor system expressing green fluorescent protein (GFP) in the presence of exogenous AHL molecules were used to detect the production of AHLs from P. aeruginosa in vivo. Mice were challenged intratracheally with alginate beads containing P. aeruginosa and E. coli and killed on different days after the challenge. By means of confocal scanning laser microscopy, GFP-expressing E. coli bacteria could be detected in the lung tissues, indicating production and excretion of AHL molecules in vivo by the infecting P. aeruginosa. AHL signals were detected mainly in lung tissues exhibiting severe pathological changes. These findings support the view that expression of AHL molecules by P. aeruginosa during infection coincides with its pathogenesis. PMID- 11021925 TI - Comparison of proteins expressed by Pseudomonas aeruginosa strains representing initial and chronic isolates from a cystic fibrosis patient: an analysis by 2-D gel electrophoresis and capillary column liquid chromatography-tandem mass spectrometry. AB - Isolates of Pseudomonas aeruginosa from chronic lung infections in cystic fibrosis (CF) patients have phenotypes distinct from those initially infecting CF patients, as well as from other clinical or environmental isolates. To gain a better understanding of the differences in these isolates, protein expression was followed using two-dimensional (2-D) gel electrophoresis and protein identification by peptide sequencing using micro-capillary column liquid chromatography-tandem mass spectrometry (microLC/MS/MS). The isolates selected for this analysis were from the sputum of a CF patient: strain 383 had a nonmucoid phenotype typical of isolates from the environment, and strain 2192, obtained from the same patient, had a mucoid phenotype typical of isolates from chronic CF lung infections. Strains 383 and 2192 were confirmed to be genetically identical by restriction endonuclease analysis, random amplified polymorphic DNA PCR, and pulsed-field gel electrophoresis. Conditions of protein extraction were optimized for consistent high-resolution separation of several hundred proteins from these clinical isolates as detected by Coomassie staining of 2-D gels. Fourteen proteins were selected for analysis; this group included those whose expression was common between both strains as well as unique for each strain. The proteins were identified by microLC/MS/MS of the peptides produced by an in-gel tryptic digestion and compared to translated data from the Pseudomonas Genome Project; optimization of this technique has allowed for the comparison of proteins expressed by strains 383 and 2192. PMID- 11021926 TI - migA, a quorum-responsive gene of Pseudomonas aeruginosa, is highly expressed in the cystic fibrosis lung environment and modifies low-molecular-mass lipopolysaccharide. AB - Pseudomonas aeruginosa is an opportunistic human pathogen which poses a major threat to patients with cystic fibrosis (CF). Excessive amounts of mucus present in the lungs of CF patients promotes the colonization of P. aeruginosa. The migA gene, encoding a putative glycosyltransferase, has been shown to be highly inducible by respiratory mucus derived from CF patients. In this study, it is further demonstrated by population transcript analysis that the migA gene is highly expressed in the CF lung environment. Deletion analysis of the migA promoter identified a las-box-like sequence commonly found in promoters that are responsive to quorum sensing regulation. Further analysis of migA expression in quorum-sensing-defective strains, as well as its expression in response to autoinducer molecules, demonstrated that migA is regulated by the RhlI/RhlR quorum sensing regulatory system. Functionally, as the MigA sequence homology data suggested, the migA gene indeed affects the structure of LPS in P. aeruginosa. Increased expression of the migA gene results in a loss of core-plus one LPS, while having no obvious effect on the long-chain O-antigen-bearing LPS. Although the exact biological role of the core-plus-one LPS is not clear, these experimental results suggest that migA up-regulation in the CF lung environment is part of the adaptive response which confers on P. aeruginosa a survival advantage. PMID- 11021927 TI - Secreted products of a nonmucoid Pseudomonas aeruginosa strain induce two modes of macrophage killing: external-ATP-dependent, P2Z-receptor-mediated necrosis and ATP-independent, caspase-mediated apoptosis. AB - A nonmucoid clinical isolate of Pseudomonas aeruginosa, strain 808, elaborated ATP-dependent and ATP-independent types of cytotoxic factors in the growth medium. These cytotoxic factors, active against macrophages, were secreted during the exponential phase of growth in a complex medium. Commensurate with the appearance of the cytotoxic activities in the cell-free growth medium, several ATP-utilizing enzymic activities, such as adenylate kinase, nucleoside diphosphate kinase and 5'-nucleotidase (ATPase and/or phosphatase), were detected in the medium. These ATP-utilizing enzymes are believed to convert external ATP, presumably effluxed from macrophages, to various adenine nucleotides, which then activate purinergic receptors such as P2Z, leading to enhanced macrophage cell death. Pretreatment of macrophages with periodate-oxidized ATP (oATP), which is an irreversible inhibitor of P2Z receptor activation, prevented subsequent ATP induced macrophage cell death. A second type of cytotoxic factor(s) operated in an ATP-independent manner such that it triggered activation of apoptotic processes in macrophages, leading to proteolytic conversion of procaspase-3 to active caspase-3. This cytotoxic factor(s) did not appear to act on procaspase-3 present in macrophage cytosolic extracts. Intact macrophages, when exposed to the cytotoxic factor(s) for 6-16 h, underwent apoptosis and demonstrated the presence of active caspase-3 in their cytosolic extracts. Interestingly, two redox proteins, azurin and cytochrome c(551), were detected in the cytotoxic preparation. When cell-line-derived or peritoneal macrophages or mast cells were incubated overnight with Q-Sepharose column flow-through fraction or with a mixture of azurin and cytochrome c(551), they underwent extensive cell death due to induction of apoptosis. PMID- 11021928 TI - Pseudomonas aeruginosa mediated apoptosis requires the ADP-ribosylating activity of exoS. AB - Pseudomonas aeruginosa is an opportunistic bacterial pathogen that primarily infects immunocompromised individuals and patients with cystic fibrosis. Using a tissue culture system, invasive strains of P. aeruginosa were discovered to induce apoptosis at high frequency in HeLa and other epithelial and fibroblast cell lines. This apoptotic phenotype in the infected cells was determined by several criteria including (i) visual changes in cell morphology, (ii) induction of chromatin condensation and nuclear marginalization, (iii) the presence of a high percentage of cells with subG1 DNA content, and (iv) activation of caspase-3 activity. Induction of the type III secretion machinery, but not invasion of P. aeruginosa is required for induction of apoptosis. The apoptosis phenotype is independent of the cytoskeletal rearrangements that occur in the host cell early after infection. Mutants in P. aeruginosa exoS fail to induce apoptosis and complementation with wild-type exoS restored the apoptosis-inducing capacity, demonstrating that ExoS is the effector molecule. Analysis of exoS activity mutants shows that the ADP-ribosylating capacity of ExoS is essential for inducing the apoptotic pathway. PMID- 11021929 TI - Role of Pseudomonas aeruginosa PhoP-phoQ in resistance to antimicrobial cationic peptides and aminoglycosides. AB - Resistance to the polycationic antibiotic polymyxin B and expression of the outer membrane protein OprH in the opportunistic pathogen Pseudomonas aeruginosa both involve the PhoP-PhoQ two-component regulatory system. The genes for this system form an operon with oprH, oprH-phoP-phoQ, that responds to Mg(2+) starvation and PhoP levels. In this study, the Mg(2+)-regulated promoter for this operon was mapped upstream of oprH by primer-extension experiments. An oprH::xylE-Gm(R) mutant H855 was constructed and measurement of the catechol 2,3-dioxygenase activity expressed from this transcriptional fusion provided evidence for a second, weak promoter for phoP-phoQ. Wild-type P. aeruginosa PAO1 strain H103 was found to exhibit Mg(2+)-regulated resistance to the alpha-helical antimicrobial cationic peptide CP28 in addition to its previously characterized resistance to polymyxin B. Resistance to this peptide was unchanged in the OprH-null mutant H855 and a PhoP-null mutant H851. In contrast, PhoQ-null mutant H854 demonstrated constitutive CP28 resistance. Northern blot analysis revealed constitutive expression of phoP in this strain, implicating PhoP-PhoQ in the resistance of P. aeruginosa to cationic peptides. Furthermore, all three null-mutant strains demonstrated increased resistance to the aminoglycoside antibiotics streptomycin, kanamycin and amikacin. Two additional mutant strains, H895 and H896, were constructed that carried unmarked deletions in oprH and were found to exhibit aminoglycoside susceptibility equivalent to that of the wild-type. This result provided definitive evidence that OprH is not involved in P. aeruginosa aminoglycoside resistance and that the changes in resistance in strain H855 and a previously reported oprH mutant were due to polar effects on phoP-phoQ rather than loss of OprH expression. A role for PhoP-PhoQ in resistance to aminoglycosides is envisaged that is distinct from that in resistance to cationic peptides and polymyxin B. PMID- 11021930 TI - Visualization of DNA-protein intermediates during activation of the Pu promoter of the TOL plasmid of Pseudomonas putida. AB - The ATP-dependent multimerization process undergone by the sigma(54)-dependent activator XylR of the TOL plasmid pWW0 of Pseudomonas putida when bound to the upstream activating sequences (UAS) of the cognate PU: promoter was examined by transmission electron microscopy (TEM). To this end, supercoiled DNA templates were combined with increasing concentrations of the constitutive XylR variant XylRDeltaA, with or without ATP or its non-hydrolysable analogue ATPgammaS, and the resulting complexes were visualized by TEM. The different types of DNA protein association were analysed and a statistical study of the frequency of the various forms was made. ATP appeared to establish an equilibrium between different molecular associations, as well as major changes in the physical shape of the DNA-protein complexes. The formation of higher nucleoprotein structures frequently bearing DNA bends became manifest. Such complexes often engaged otherwise separated UAS-containing plasmids, indicating that the ATP-driven multimer included XylR molecules recruited in trans. Whilst ATP caused the different types of XylR-DNA complex to occur at quite balanced frequencies, ATPgammaS appeared to displace the distribution predominantly towards the higher order forms. These data are compatible with the notion that each time ATP is hydrolysed the transcriptional activation complex is disassembled. PMID- 11021931 TI - Activation of Pseudomonas aeruginosa elastase in Pseudomonas putida by triggering dissociation of the propeptide-enzyme complex. AB - The propeptide of Pseudomonas aeruginosa elastase functions both as an intramolecular chaperone required for the folding of the enzyme and as an inhibitor that prevents activity of the enzyme before its secretion into the extracellular medium. Since expression of the lasB gene, which encodes elastase, in Pseudomonas putida did not result in extracellular elastase activity, it has been suggested that the enzyme is not recognized by the Xcp secretion machinery of the heterologous host. Here, it is demonstrated that the proenzyme is normally processed in P. putida and that it is indeed not actively secreted by the Xcp machinery. Nevertheless, substantial amounts of the enzyme were detected in the extracellular medium. Co-immunoprecipitations revealed that the extracellular enzyme was associated with the propeptide, which explains the lack of enzymic activity. Since the propeptide-enzyme complex in P. putida apparently does not dissociate spontaneously, it is concluded that a host-specific factor is required to induce this event. Mutants were selected which showed extracellular elastase activity. Two mutations, located within the lasB gene, were further characterized. These mutations, resulting in the substitution of Ala and Thr at positions -15 and -153, respectively, of the propeptide (where position +1 is defined as the first residue of the mature enzyme) destabilized the propeptide enzyme complex. It is concluded that Ala-15 and Thr-153 are required for the inhibitor function, but not for the chaperone function of the propeptide. PMID- 11021933 TI - Cell-associated degradation affects the yield of secreted engineered and heterologous proteins in the Bacillus subtilis expression system. AB - A series of chimeric alpha-amylase genes derived from amyL, which encodes the liquefying alpha-amylase from Bacillus licheniformis, were constructed in vitro using gene splicing techniques. The gene constructs were cloned in Bacillus subtilis, where their ability to direct the synthesis and secretion of active alpha-amylase was determined. Detectable alpha-amylase activity was observed for some, but not all, of the chimeric proteins. Studies on the secretion of wild type AmyL and its chimeric derivatives revealed that, whilst these proteins were stable in the extracellular milieu, all were subject to some degree of degradation during secretion. The chimeric enzymes were degraded to a greater extent than the native enzyme. These findings suggest that cell-associated proteolysis is a significant problem affecting the use of B. subtilis as host bacterium for the production of heterologous proteins. PMID- 11021932 TI - The carboxyl terminus of the Bacillus subtilis SecA is dispensable for protein secretion and viability. AB - The Escherichia coli secretion-dedicated chaperone SecB targets a subset of proteins to the translocase by interacting with the carboxyl (C-) terminus of SecA. This region of SecA is highly conserved in Eubacteria, but despite its presence in the Bacillus subtilis SecA, the B. subtilis genome does not appear to contain a gene for a clear homologue of SecB. Deletion of the C-terminus of the B. subtilis SecA yields cells that have normal viability, but that exhibit a response reminiscent of oxidative stress and the loss of a number of secretory proteins from the culture supernatant. Semi-quantitative RT-PCR demonstrates that these proteins are expressed at lower levels. The C-terminus of SecA fused to glutathione S:-transferase (GST) specifically binds a cytosolic protein, termed MrgA. This protein has been reported to function in relation to oxidative stress, but deletion of the mrgA gene does not result in a secretion defect nor does it cause an oxidative stress response. It is concluded that the C-terminus of the B. subtilis SecA is not essential for secretion and viability. PMID- 11021934 TI - Expression of the ftsY gene, encoding a homologue of the alpha subunit of mammalian signal recognition particle receptor, is controlled by different promoters in vegetative and sporulating cells of Bacillus subtilis. AB - Bacillus subtilis FtsY (Srb) is a homologue of the alpha subunit of the receptor for mammalian signal-recognition particle (SRP) and is essential for protein secretion and vegetative cell growth. The ftsY gene is expressed during both the exponential phase and sporulation. In vegetative cells, ftsY is transcribed with two upstream genes, rncS and smc, that are under the control of the major transcription factor sigma(A). During sporulation, Northern hybridization detected ftsY mRNA in wild-type cells, but not in sporulating cells of sigma(K) and gerE mutants. Therefore, ftsY is solely expressed during sporulation from a sigma(K)- and GerE-controlled promoter that is located immediately upstream of ftsY inside the smc gene. To examine the role of FtsY during sporulation, the B. subtilis strain ISR39 was constructed, a ftsY conditional mutant in which ftsY expression can be shut off during spore formation but not during the vegetative state. Electron microscopy showed that the outer coat of ISR39 spores was not completely assembled and immunoelectron microscopy localized FtsY to the inner and outer coats of wild-type spores. PMID- 11021935 TI - The Vibrio cholerae O1 chromosomal integron. AB - Until the discovery of the Vibrio cholerae repeat (VCR), the gene capture and expression systems termed integrons had been typically associated with antibiotic resistance gene cassettes with usually less than five genes in an array. A method is described for the cloning of the ends of large cassette arrays. Conserved restriction sites within VCRs facilitated the mapping by Southern hybridization and cloning of the 5' end of the VCR array, and using appropriate fragments it was possible to develop a physical map of the region of the V. cholerae chromosome. Sequence determination of the predicted beginning of this region revealed intI4, a member of the integron family of integrases. Comparison of these sequences from El Tor, Classical and serotype O134 V. cholerae strains identified the 3' end of the attI site, thereby defining the class 4 integron in one of the V. cholerae chromosomes, and providing the first evidence for integron like site-specific recombination within V. cholerae. Conduction assays demonstrated IntI1-mediated recombination between VCRs. Restriction mapping places the sequences of intI4 and 26 VCR gene cassettes in arrays within a 120 kb region of the V. cholerae O1 strain 569B genome. This region contains an estimated 150 VCR gene cassettes, dwarfing previously described arrays. Southern analysis of genomic DNA from strains of Vibrio anguillarum, Vibrio mimicus and a number of V. cholerae serotypes revealed fragments that hybridized with VCR specific probes but showed a high degree of restriction fragment length polymorphism. These data facilitate the identification of part of a new class 5 integron from V. mimicus. PMID- 11021936 TI - Genetic relationships between clinical and environmental Vibrio cholerae isolates based on multilocus enzyme electrophoresis. AB - A total of 107 isolates of Vibrio cholerae, including 29 strains belonging to serogroup O139, were studied using multilocus enzyme electrophoresis (MLEE) to determine allelic variation in 15 housekeeping enzyme loci. All loci were polymorphic and 99 electrophoretic types (ETs) were identified from the total sample. No significant clustering of isolates was detected in the dendrogram generated from a matrix of coefficients of distances with respect to serogroup, biotype or country of isolation. The mean genetic diversity of this V. cholerae population (H:=0.50) was higher than reported previously. Linkage disequilibrium analysis of the MLEE data showed a clonal structure for the entire population, but not in some of the population subgroups studied. This suggests an epidemic population structure. The results showed that the O139 strains were not clustered in a unique ET, in contrast to previous MLEE studies. This higher genetic variation of the O139 serogroup is concordant with ribotyping studies. The results also confirm that the O139 and O1 ElTor isolates are genetically more closely related to each other than to all the other subpopulations of V. cholerae studied. PMID- 11021937 TI - Genome size and genetic map of Cowdria ruminantium. AB - Cowdria ruminantium is the cause of a serious tick-borne disease of domestic ruminants, known as heartwater or cowdriosis. The organism belongs to the tribe Ehrlichieae:, which contains obligate intracellular pathogens, causing several important animal and human diseases. Although a few C. ruminantium genes have been cloned and sequenced, very little is known about the size, gross structure and organization of the genome. This paper presents a complete physical map and a preliminary genetic map for C. ruminantium. Chromosomal C. ruminantium DNA was examined by PFGE and Southern hybridization. PFGE analysis revealed that C. ruminantium has a circular chromosome approximately 1576 kb in size. A physical map was derived by combining the results of PFGE analysis of DNA fragments resulting from digestion of the whole genome with KSP:I, RSR:II and SMA:I and Southern hybridization analysis with a series of gene probes and isolated macrorestriction fragments. A genetic map for C. ruminantium with a mean resolution of 290 kb was established, the first for a member of the Ehrlichieae: A total of nine genes or cloned C. ruminantium DNA fragments were mapped to specific KSP:I, RSR:II and SMA:I fragments, including the major antigenic protein gene, map-1. PMID- 11021938 TI - The nitric oxide regulated nor promoter of Paracoccus denitrificans. AB - The promoter of the Paracoccus denitrificans nitric oxide reductase operon (norCBQDEF) has been characterized by primer extension and deletion analysis. A major transcript that is detectable only in anaerobically grown cells initiates 43.5 bp downstream of the centre of a putative binding site for the transcription factor NNR (nitrite and nitric oxide reductase regulator, which is known to regulate nor expression). A minor transcript initiates 121 bp upstream of the major transcript and is detectable in cells grown aerobically or anaerobically. Deletion derivatives of the nor promoter region were constructed and analysed in vivo using transcriptional fusions to the reporter gene lacZ. Expression patterns from promoter deletions in a wild-type strain and an nnr mutant confirmed that the minor transcript is NNR independent, and makes a small contribution to nor expression under both aerobic and anaerobic growth conditions. A deletion derivative truncated to within 7 bp of the putative NNR-binding site showed a near wild-type response to anaerobic growth, showing that no upstream DNA sequences are required for activation of the major promoter. Site-directed mutagenesis of the putative NNR-binding site confirmed that this is the major cis acting sequence mediating the anaerobic inducibility of nor expression. PMID- 11021939 TI - T4 early promoter strength probed in vivo with unribosylated and ADP-ribosylated Escherichia coli RNA polymerase: a mutation analysis. AB - The consensus sequence of T4 early promoters differs in length, sequence and degree of conservation from that of Escherichia coli sigma(70) promoters. The enzyme interacting with these promoters, and transcribing the T4 genome, is native host RNA polymerase, which is increasingly modified by the phage-encoded ADP-ribosyltransferase, Alt. T4 early transcription is a very active process, possibly out-competing host transcription. The much stronger T4 promoters enhance viral transcription by a factor of at least two and the Alt-catalysed ADP ribosylation of the host enzyme triggers an additional enhancement, again by a factor of about two. To address the question of which promoter elements contribute to the increasing transcriptional activity directed towards phage genes, the very strong E. coli promoter, Ptac, was sequentially mutated towards the sequence of the T4 early promoter consensus. Second, mutations were introduced into the highly conserved regions of the T4 early promoter, P8.1. The co-occurrence of the promoter-encoding plasmid pKWIII and vector pTKRI, which expresses Alt in E. coli, constitutes a test system that allows comparison of the transcriptional activities of phage and bacterial promoters, in the presence of native, or alternatively ADP-ribosylated RNA polymerase. Results reveal that T4 early promoters exhibit a bipartite structure, capable of strong interaction with both types of RNA polymerase. The -10, -16, -42 and -52 regions are important for transcript initiation with the native polymerase. To facilitate acceleration of transcription, the ADP-ribosylated enzyme requires not only the integrity of the 10, -16 and -35 regions, but also that of position -33, and even more importantly, maintenance of the upstream promoter element at position -42. The latter positions introduced into the E. coli Ptac promoter render this mutant promoter responsive to Alt-ADP-ribosylated RNA polymerase, like T4 early promoters. PMID- 11021940 TI - A mutation which affects both the specificity of PtsG sugar transport and the regulation of ptsG expression by Mlc in Escherichia coli. AB - Normally glucosamine (GlcN) is not a substrate for EIICB(Glc) of the glucose phosphotransferase system (PTS), encoded by ptsG, but it is transported by the mannose (Man) PTS, encoded by manXYZ. A mutation, umgC, has been described in Escherichia coli which allows a strain mutated in the Man PTS to grow on GlcN. The umgC mutation was mapped to the ptsG region and was proposed to make ptsG expression constitutive. Transcription of ptsG is regulated by the repressor Mlc so that mutations in mlc enhance the expression of ptsG. An mlc mutation, however, is not sufficient to allow good growth on GlcN, unlike the umgC mutation. The umgC mutation is shown to enhance expression of ptsG even in the absence of any PTS sugar transport, but the increase is greater in the presence of GlcN or Man. The umgC mutation also increases expression of the ptsHI and manXYZ operons, which are both regulated by Mlc. The umgC mutation was sequenced and two mutations were found: one, G176D, within the IIC membrane domain and the second, E472K, within the soluble IIB domain of PtsG. The cloned UmgC allele shows the enhanced transport and regulatory characteristics of the chromosomal mutation. Analysis of the two mutations present individually on plasmids shows that the IIC mutation is responsible for both the effect on sugar specificity and regulation. PMID- 11021942 TI - Intracellular pH determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis. AB - Intracellular pH (pH(i)) is an essential parameter in the regulation of intracellular processes. Thus, its measurement might provide clues regarding the physiological state of cells cultivated in vitro. pH(i) of the filamentous, pristinamycin-producing Streptomyces pristinaespiralis was determined by epifluorescence microscopy and image analysis using the pH-sensitive fluorescent probe BCECF-AM [2', 7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, acetoxymethyl ester]. Staining cell culture samples (OD(660)=1) of S. pristinaespiralis with 20 microM BCECF at 28 degrees C for 30 min yielded a green/red fluorescence ratio (R:(527/600)) that correlated with the pH(i) of the cells for values ranging from 6.5 to 8.5. When S. pristinaespiralis was cultivated in pristinamycin-producing conditions (in batch mode, with a constant external pH of 6.8), the measured pH(i) varied between 6.3 and 8.7. In fact, pH(i) correlated with the excretion of pristinamycins and glucose consumption during the production process. PMID- 11021941 TI - Antisense PNA effects in Escherichia coli are limited by the outer-membrane LPS layer. AB - Antisense peptide nucleic acids (PNAs) can inhibit Escherichia coli gene expression and cell growth through sequence-specific RNA binding, and this opens possibilities for novel anti-infective agents and tools for microbial functional genomics. However, the cellular effects of PNAs are limited relative to effects in cell extracts, presumably because of cell barrier components such as the outer membrane lipopolysaccharide (LPS) layer or drug efflux pumps, both of which function to exclude antibiotics and other foreign molecules. To evaluate the importance of such cellular factors on PNA effects, the authors developed a positive assay for antisense inhibition by targeting the lac operon repressor and compared PNA susceptibilities in mutant and wild-type E. coli by assessing lacZ induction. Strains with defective LPS (AS19 and D22) were more permeable to the antibiotic nitrocefin and more susceptible to PNA than the wild-type. Also, PNA potency was improved in wild-type cells grown in the presence of certain cell wall-permeabilizing agents. In contrast, the activities of the Acr and Emr drug efflux pumps were not found to affect PNA susceptibility. The results show that the LPS layer is a major barrier against cell entry, but PNAs that can enter E. coli are likely to remain active inside cells. PMID- 11021943 TI - Dioctyl phthalate increases the percentage of unsaturated fatty acids with a concomitant decrease in cellular heat shock sensitivity in the yeast Saccharomyces cerevisiae. AB - In the past it has been reproducibly demonstrated that 37 degrees C-grown DBY747 yeast cells have 29% more unsaturated fatty acids and a 3 degrees C higher maximal heat shock response (HSR) than their 25 degrees C counterparts. Suddenly the HSR and lipid profiles of cells grown at 25 degrees C and 37 degrees C became indistinguishable from one another. This paper reports an aberrantly high level of unsaturated fatty acids and an abnormally insensitive HSR in cells grown at 25 degrees C in yeast nitrogen base (YNB) that has been reconstituted from dehydrated medium packaged in 'new' plastic containers. Effective even at a 1:600 dilution of reconstituted medium in laboratory-made YNB, the 'active ingredient' was identified using a combination of HPLC and mass spectroscopy as dioctyl phthalate (a plasticising agent). Furthermore, the same levels of increase in the percentage of unsaturated fatty acids and decrease in the sensitivity of HSR were found in cells grown in laboratory-made YNB that contained as little as 36 microM pure dioctyl phthalate. This compound nevertheless failed to elicit an observable effect on cellular growth rate at levels up to and including 144 microM. These results suggest that dioctyl phthalate causes yeast cells to accumulate high levels of unsaturated fatty acids with a concomitant decrease in the sensitivity of the HSR, without compromising overall cellular function. They also support earlier work that suggested that the HSR is exquisitely sensitive to the level of unsaturated fatty acids present in yeast cells. PMID- 11021944 TI - Mitochondrial respiratory mutants of Saccharomyces cerevisiae accumulate glycogen and readily mobilize it in a glucose-depleted medium. AB - Mutant strains of Saccharomyces cerevisiae defective in respiration have been reported to be unable to store glycogen, as revealed by the iodine-staining method. In this report, it is shown that in contrast to this claim, mitochondrial respiratory mutants accumulated even more glycogen than wild-type cells during the fermentative growth on glucose. However, as soon as glucose was exhausted in the medium, these mutants readily and completely mobilized their glycogen content, contrary to wild-type cells which only transiently degraded this polymer. The mobilization of glycogen was a specific trait resulting from a defect in mitochondrial function that could not be suppressed by mutations in the cAMP- and Pho85 protein kinase-dependent nutrient-sensing pathways, and by other mutations which favour glycogen synthesis. To account for this mobilization, it was found that respiration-defective cells not only contained a less active glycogen synthase, but also a more active glycogen phosphorylase. Since glucose 6 phosphate (Glc6P) is a potent inhibitor of the phosphorylation and an activator of the dephosphorylation processes of glycogen synthase and glycogen phosphorylase, it is suggested that the drop in Glc6P observed at the onset of glucose depletion in respiration-deficient cells triggers this rapid and sustained glycogen mobilization. It is also proposed that this degradation provides the energy for the viability of respiratory mutants in glucose-starved medium. PMID- 11021945 TI - The Aspergillus nidulans cysA gene encodes a novel type of serine O acetyltransferase which is homologous to homoserine O-acetyltransferases. AB - The Aspergillus nidulans cysA gene was cloned by functional complementation of the cysA1 mutation that impairs the synthesis of O:-acetylserine. The molecular nature of cysA1 and cysA103 alleles was characterized; a nucleotide substitution and a frame shift were found in the former and a deletion mutation in the latter. The CYSA protein is 525 amino acids long and is encoded by an uninterrupted open reading frame. Expression of the cysA gene appears not to be regulated by sulfur, carbon and nitrogen sources. Protein sequence analysis reveals extensive similarity to homoserine O:-acetyltransferases, particularly the bacterial ones, and no homology with known serine O:-acetyltransferases. The authors propose that the CYSA protein is analogous to serine O:-acetyltransferases, i.e. it catalyses the same reaction but has an independent evolutionary origin. PMID- 11021946 TI - Mannitol-1-phosphate dehydrogenase from Cryptococcus neoformans is a zinc containing long-chain alcohol/polyol dehydrogenase. AB - Cryptococcus neoformans, the causative agent of cryptococcosis, produces large amounts of mannitol in culture and in infected mammalian hosts. Although there is considerable indirect evidence that mannitol synthesis may be required for wild type stress tolerance and virulence in C. neoformans, this hypothesis has not been tested directly. It has been proposed that mannitol-1-phosphate dehydrogenase (MPD) is required for fungal mannitol synthesis, but no MPD deficient fungal mutants or cDNAs or genes encoding fungal MPDs have been described. Therefore, C. neoformans was purified from a 148 kDa homotetramer of 36 kDa subunits that catalysed the reaction mannitol1-phosphate+NAD--><--fructose 6-phosphate+NADH. Partial peptide sequences were used to isolate the corresponding cDNA and gene, and the deduced MPD protein was found to be homologous to the zinc-containing long-chain alcohol/polyol dehydrogenases. Lysates of Saccharomyces cerevisiae transformed with the cDNA of interest (but not vector-transformed controls) contained MPD catalytic activity. Lastly, Northern analyses demonstrated MPD mRNA in glucose- and mannitol-grown C. neoformans cells. Thus, MPD has been purified and characterized from C. neoformans, and the corresponding cDNA and gene (MPD1) cloned and sequenced. Availability of C. neoformans MPD1 should permit direct testing of the hypotheses that (i) MPD is required for mannitol biosynthesis and (ii) the ability to synthesize mannitol is essential for wild-type stress tolerance and virulence. PMID- 11021948 TI - Sustainability of Canadian medicare: Optimism for the future (part I). PMID- 11021947 TI - Dependence of Trichomonas vaginalis upon polyamine backconversion. AB - Trichomonas vaginalis grown for 16 h in the presence of [(14)C]spermine formed a high intracellular pool of [(14)C]spermidine and a small but detectable pool of [(14)C]putrescine. When [(3)H]putrescine was added to the growth medium, a large intracellular pool of [(3)H]putrescine was found, but it was not further metabolized, confirming previous studies suggesting the absence of a forward directed polyamine synthetic pathway in T. vaginalis. Spermidine:spermineN:(1) acetyltransferase (SSAT) and polyamine oxidase enzyme activities were detected which collectively converted spermine to spermidine. Polyamine oxidase was localized in the hydrogenosome-enriched fraction, whereas SSAT was found predominantly in the cytosolic fraction. In the presence of saturating substrate, the trichomonad SSAT had an activity of 0. 39+/-0.09 nmol min(-1) (mg protein)( 1) (the mean of five analyses) and an apparent K:(m) for spermine of 1.7 microM. The enzyme was competitively inhibited by di(ethyl)norspermine with a K:(i) of 28 microM. Growth studies indicated that 50 microM di(ethyl)norspermine caused a 68% and 84% reduction in the intracellular concentrations of spermidine and spermine, respectively. The trichomonad polyamine oxidase required FAD as a cofactor and had an apparent K:(m) of 6.0 microM for N(1)-acetylspermine. The potential of bis(alkyl) polyamine analogues as antitrichomonad agents is discussed. PMID- 11021949 TI - Influenza immunization for high risk populations. PMID- 11021950 TI - Hunting on the medical savannah. PMID- 11021951 TI - Message from the president. PMID- 11021952 TI - Canadian national high blood pressure prevention and control strategy. AB - Despite major efforts to prevent and control high blood pressure, it is one of the most common and important health problems facing Canadians. To address this issue, Health Canada, in collaboration with the Canadian Coalition for High Blood Pressure Prevention and Control, established an Expert Working Group to prepare a national strategy. The present report outlines a strategy to prevent and control high blood pressure. It is directed at policy makers at the local, provincial, and/or territorial and national levels in both the health and nonhealth sectors. The strategy is based on current research and expertise. A multifaceted, comprehensive approach is proposed because there is no one intervention that will accomplish the goal of improving the health of Canadians through high blood pressure prevention and control. The present report focuses on the general population. It does not address the unique needs of children, pregnant women or aboriginal peoples. Each of these groups needs to be studied in its own right, and, in particular, with the involvement of aboriginal people themselves. An implementation committee has been established to realize this strategy, and the Canadian Hypertension Society is a key stakeholder in this effort. Several initiatives are underway. Strong advocates are necessary to increase public awareness and to support the system changes required for a successful public health approach to reduce the prevalence of hypertension and its complications. PMID- 11021953 TI - A novel process for updating recommendations for managing hypertension: rationale and methods. AB - BACKGROUND: There are numerous hypertension consensus recommendations intended for practising physicians. However, recommendations in their current format have limited impact on improving hypertension control. MATERIALS AND METHODS: A group of national societies, headed by the Canadian Hypertension Society, the Heart and Stroke Foundation of Canada, the Canadian Coalition for High Blood Pressure Prevention and Control, and Health Canada has developed strategies to maintain annually updated recommendations for hypertension management and to provide greater opportunities for their implementation into clinical practice. The process is overseen by a steering committee. Subcommittees have been formed for each of a list of topics seen as important to the control of hypertension. The subcommittees, with the aid of a central librarian, conduct annual literature reviews in accordance with Cochrane Collaboration strategies. Modified existing and new recommendations are forwarded to a group with expertise in clinical epidemiology. Grades of evidence are assigned to each recommendation. Revised recommendations based on the above process will be presented annually at the conjoint Canadian Hypertension Society/Canadian Cardiovascular Congress meeting. Under the leadership of the Cardiovascular Disease Division of the Laboratory Centre for Disease Control, Health Canada, a committee has been charged with the implementation process. CONCLUSIONS: The improvements of the current process over previous national hypertension recommendations are four-fold. First, the recommendations will be updated annually. Second, the methodology has been improved. Third, the grading system can be used in the evaluation of complex study designs. Finally, the implementation process is extended. The authors are optimistic that these changes will contribute to the improvement of hypertension control in the Canadian population. PMID- 11021954 TI - Glucose and insulin metabolism are important determinants of left ventricular mass in obese hypertensives. AB - BACKGROUND: Obesity and hypertension, the major modifiable clinical determinants of left ventricular mass, are both associated with a state of insulin resistance. OBJECTIVE: To determine the relationships between glucose and insulin metabolism and left ventricular mass in a group of obese, nondiabetic, hypertensive people. PATIENTS AND METHODS: Twenty-two obese, nondiabetic, hypertensive people (10 men), free of coronary or valvular heart disease, with normal left ventricular function were studied. The mean age was 55+/-12 years, body mass index 32.8+/-4.8 kg/m2, and systolic and diastolic blood pressures 149.0+/-11.0 mmHg and 98.0+/ 4.0 mmHg, respectively. Left ventricular mass corrected for height (LVM/Ht) and glucose and insulin metabolism (3 h intravenous glucose tolerance test) were measured after a four- to six-week washout period of any antihypertensive medication. RESULTS: The mean LVM/Ht was 119.5+/-11.9 kg/m. The following metabolic measures correlated with LVM/Ht in a univariate analysis: total insulin integration area (r=0.54, P=0.008); fasting insulin (r=0.43, P=0.04); insulin at 90 min (r=0.54, P=0.013); and peak glucose levels (r=0.51, P=0.013). Stepwise multivariate linear regression analysis showed that only total insulin integration area (P=0.005) and peak glucose levels (P=0.01) correlated with LVM/Ht. CONCLUSION: In obese, nondiabetic, hypertensive people, left ventricular mass is associated with circulating glucose and insulin levels. PMID- 11021955 TI - Patterns of amlodipine and felodipine use in an elderly Quebec population. AB - OBJECTIVES: To assess drug prescription patterns and medical resource consumption in an elderly population in Quebec receiving amlodipine or felodipine for the treatment of hypertension. PATIENTS AND METHODS: Sociodemographic, clinical and drug claim data for a random sample of hypertensive patients 65 years of age and older with at least one claim for amlodipine or felodipine between August 1, 1990 and August 31, 1997 were extracted from the Regie de l'assurance maladie du Quebec (RAMQ) database. Patterns of prescription renewal, drug switch and compliance rates, and health care resource use were established for both an amlodipine and a felodipine group. Long term persistence on treatment was quantified by survival curve analysis. RESULTS: The amlodipine (5188 patients) and felodipine (2630 patients) groups were similar in terms of sex ratio (66.7% female) and age (mean 74 years). Average compliance rates for amlodipine patients (67.9%) were significantly higher than for felodipine patients (66.2%) (P<0.01), and switch rates were 5.4-fold higher in the latter group. Patients initiating treatment with felodipine had a 27% increased rate of discontinuation (relative risk 1.27) compared with the amlodipine patients. In addition, patients with at least one year of follow-up data were more likely to maintain amlodipine as part of their antihypertensive regimens than felodipine. After adjustment, medical resource consumption patterns were similar for both groups except for an increase in the number of specialist visits for the amlodipine treatment group. CONCLUSIONS: Patients who received amlodipine, either as a monotherapy or as part of a multitherapy regimen, were more compliant and persistent with their treatment than patients on felodipine. The data suggest that amlodipine may provide more effective long term hypertension control than felodipine, and that the two drugs are not therapeutically equivalent. PMID- 11021956 TI - A prospective, randomized, open-label trial comparing telmisartan 80 mg with valsartan 80 mg in patients with mild to moderate hypertension using ambulatory blood pressure monitoring. AB - OBJECTIVE: To compare the antihypertensive efficacy and tolerability of telmisartan 80 mg with valsartan 80 mg throughout a 24 h dosing interval. DESIGN: A prospective, randomized, open-label, blinded end point, parallel group study. Treatment efficacy was compared using ambulatory blood pressure monitoring (ABPM), cuff sphygmomanometry and calculated responder rates. Tolerability was assessed by physical examination, laboratory parameters, 12-lead electrocardiogram, blood pressure and heart rate monitoring, and evaluation of adverse events. SETTING: Thirty-five centres in the United States. PATIENTS: Four hundred and twenty-six patients with mild to moderate essential hypertension entered the study. Ninety-two per cent (n=393) completed the study. INTERVENTIONS: Patients underwent a four-week, single-blind, placebo run-in period before being randomly assigned to once-daily oral telmisartan 80 mg (n=214) or valsartan 80 mg (n=212) for an eight-week, open-label treatment period. RESULTS: Treatment with telmisartan was associated with a significantly greater mean reduction from baseline in the last 6 h ABPM mean for diastolic blood pressure compared with the valsartan-treated group (-7.5+/-0.6 mmHg versus 5.2+/-0.6 mmHg, respectively, P<0.01). Secondary analyses showed significantly greater efficacy with telmisartan 80 mg than with valsartan 80 mg, including greater mean reductions from baseline of ABPM (systolic blood pressure and diastolic blood pressure) during the daytime (06:00 to 21:59) and morning (06:00 to11:59) hours, and larger decreases in trough cuff blood pressure (P<0.01). Both treatments showed placebo-like tolerability profiles. CONCLUSIONS: Telmisartan 80 mg once daily was superior to valsartan 80 mg once daily in reducing diastolic blood pressure during the last 6 h of the 24 h dosing interval. These results may be due to telmisartan's longer plasma half-life or to a higher potency compared with valsartan, such that a higher dose of valsartan may produce effects similar to those of 80 mg telmisartan. These data confirm the long duration of action of telmisartan with consistent and sustained control of blood pressure over 24 h and during the last 6 h of the dosing interval. Both treatments were well tolerated; the adverse event data confirmed the excellent tolerability profiles of telmisartan and valsartan that have been reported previously. PMID- 11021957 TI - Molecular and cellular mechanisms regulating vascular function and structure- implications in the pathogenesis of hypertension. AB - Human essential hypertension is a complex, multifactorial, quantitative trait under polygenic control. The fundamental hemodynamic abnormality in hypertension is increased peripheral resistance due primarily to changes in vascular structure and function. These changes include arterial wall thickening and abnormal vascular tone, and are due to alterations in the biology of the cellular and noncellular components of the arterial wall. Multiple interacting humoral and mechanical factors as well as oxidative stress stimulate complex signalling pathways, which modulate vascular smooth muscle cell contraction and growth. Under normal physiological conditions, these finely regulated processes maintain vessel wall integrity and prevent pathological increases in blood pressure. However, under abnormal conditions, increased humoral and mechanical signalling results in vascular wall thickening and increased vascular tone, which play an important role in the pathogenesis and maintenance of hypertension. The present review discusses recent developments in the understanding of cellular and molecular mechanisms underlying vascular regulation in hypertension. PMID- 11021958 TI - Guidelines for the management of hypertension. AB - In Canada, as in all industrialized countries, hypertension is the most common indication for visits by adults to doctors, and is becoming even more prevalent largely as a consequence of two factors--the increasing age and the increasing obesity of the population. The control of hypertension in Canada and elsewhere is woefully inadequate. Recent guidelines from the United States and Canada emphasize similar principles designed to improve control. These include the assessment of overall cardiovascular risk, careful measurements of blood pressure, effective use of lifestyle modifications, targeted antihypertensive drug therapy and various manoeuvres to improve long term adherence to therapy. Cardiologists will play an increasingly important role in managing hypertension as a way to reduce the cardiovascular diseases that remain the leading causes of morbidity and mortality. PMID- 11021959 TI - A comparative review of the efficacy of antihypertensive agents on 24 h ambulatory blood pressure. AB - BACKGROUND: Ambulatory blood pressure (ABP) monitoring has contributed significantly to the diagnosis, prognosis and treatment of hypertension. However, most of the published reviews have not focused on the 24 h efficacy of conventional and new antihypertensive agents as measured by ABP monitoring. OBJECTIVES: To discuss the importance of 24 h blood pressure control, and to review the antihypertensive efficacy of conventional antihypertensive agents and of angiotensin II type I receptor (AT1R) blockers as assessed by ABP monitoring. PATIENTS AND METHODS: The 24 h antihypertensive efficacy of the different classes of agents was reviewed based on clinical, randomized, double-blind trials published in peer review journals. These trials were mainly performed in the authors' research unit but were performed in other centres as well. RESULTS AND CONCLUSIONS: Most antihypertensive agents provided 24 h ABP control. However, in the authors' experience, hydrochlorothiazide, some angiotensin converting enzyme inhibitors and low dosages of the calcium antagonist diltiazem did not permit satisfactory blood pressure reduction as assessed by ABP monitoring in truly hypertensive patients. AT1R blockers and especially those of the new generation characterized by tight and long lasting AT1-receptor binding produced adequate 24 h ABP control and continued effectiveness during the high risk early morning hours. PMID- 11021960 TI - Issue on hypertension. PMID- 11021961 TI - Relevance of the force-velocity relationship in the activation of mono- and Bi articular muscles in slow arm movements in humans. AB - We have investigated whether differences in EMG activity in mono- and bi articular muscles for concentric and eccentric contractions (van Bolhuis, Gielen, & van Ingen Schenau, 1998) have to be attributed to a specific muscle coordination strategy or whether they are merely a demonstration of adaptations necessary to adjust for muscle contractile properties. Slow, multi-joint arm movements were studied in a horizontal plane with an external force applied at the wrist. Kinematics and electromyography data from 10 subjects were combined with data from a 3-D model of the arm and a Hill-type muscle model. Data for both mono- and bi-articular muscles revealed a higher activation in concentric than in eccentric contractions. The model calculations indicated that the measured difference in activation (20%) was much larger than expected based on the force velocity relationship (predicting changes of approximately 5%). Although these findings eliminate the force-velocity relationship as the main explanation for changes in EMG, it cannot be ruled out that other muscle contractile properties, such as history dependence of muscle force, determine muscle activation levels in the task that was studied. PMID- 11021962 TI - Preoperative endocrine therapy for breast cancer. AB - The preoperative use of systemic therapy for primary breast cancer has the potential to downstage tumours. This would render suitable for breast conservation some tumours that were unsuitable at initial presentation, or would convert some inoperable locally advanced breast cancers into tumours that are operable. No survival benefit has been demonstrated for neoadjuvant chemotherapy compared with the same therapy given in an adjuvant setting. Preoperative endocrine therapy, in contrast to neoadjuvant chemotherapy, has fewer side effects and has the potential additional advantage that it can be continued throughout the perioperative period. Current data have shown that, in patients with an oestrogen receptor (ER)-positive tumour, a response approaching 70% could be reached in approximately 3 months using traditional endocrine manipulation such as tamoxifen. Randomised clinical trials are warranted to demonstrate the superiority of preoperative endocrine therapy over conventional adjuvant endocrine therapy, to define the optimum duration of therapy, and to identify the best endocrine agents. Both clinical and laboratory studies are also required to identify factors (in addition to ER) that would precisely predict the response and hence to select appropriate patients and to improve existing methods of monitoring response. PMID- 11021963 TI - c-Myc in breast cancer. AB - Ever since Bishop and his co-workers discovered the c-myc gene in the late 1970s (Bishop 1982), voluminous literature has documented its central role in proliferation and malignant transformation of human and animal cells (Amati et al. 1998, Bouchard et al. 1998, Dang et al. 1999). Most, if not all, types of human malignancy have been reported to have amplification and/or overexpression of this gene, although the frequency of these alterations varies greatly among different reports (Nesbit et al. 1999). In 1992, researchers started to realize that aberrant expression of c-myc could cause apoptosis (Evan et al. 1992, Shi et al. 1992), although the phenomenon had actually been observed much earlier (Wurm et al. 1986). Studies in recent years have further shown that the c-myc gene regulates growth, both in the sense of cell size and in the context of tissue differentiation (Gandarillas & Watt 1997, Iritani & Eisenman 1999, Johnston et al. 1999, Schmidt 1999, Schuhmacher et al. 1999). Thus, it is now known that the c-myc gene participates in most aspects of cellular function, including replication, growth, metabolism, differentiation, and apoptosis (Packham & Cleveland 1995, Hoffman & Liebermann 1998, Dang 1999, Dang et al. 1999, Elend & Eilers 1999, Prendergast 1999). How the c-Myc protein may be specifically directed to perform one, but not the others, of these functions is still obscure, despite the fact that the relevant literature has been accumulating at a fast pace in the past two decades. This review focuses on the profound roles of c-Myc in breast cancer and in the actions of the hormones that are eitologically related to breast cancer. PMID- 11021964 TI - Fibroblast growth factors, their receptors and signaling. AB - Fibroblast growth factors (FGFs) are small polypeptide growth factors, all of whom share in common certain structural characteristics, and most of whom bind heparin avidly. Many FGFs contain signal peptides for secretion and are secreted into the extracellular environment, where they can bind to the heparan-like glycosaminoglycans (HLGAGs) of the extracellular matrix (ECM). From this reservoir, FGFs may act directly on target cells, or they can be released through digestion of the ECM or the activity of a carrier protein, a secreted FGF binding protein. FGFs bind specific receptor tyrosine kinases in the context of HLGAGs and this binding induces receptor dimerization and activation, ultimately resulting in the activation of various signal transduction cascades. Some FGFs are potent angiogenic factors and most play important roles in embryonic development and wound healing. FGF signaling also appears to play a role in tumor growth and angiogenesis, and autocrine FGF signaling may be particularly important in the progression of steroid hormone-dependent cancers to a hormone independent state. PMID- 11021965 TI - Charge pair interactions in a model transmembrane helix in the ER membrane. AB - We have examined the effects of Lys-Asp charge pair interactions on the position of a model poly-Leu transmembrane helix in the ER membrane using the so-called "glycosylation mapping" technique. Based on an analysis of a set of constructs containing pairs of positively charged Lys and negatively charged Asp residues in various positions in the model helix, we show that the helix is located deeper in the membrane when Lys and Asp are placed one helical turn apart than for other spacings of the two residues. These results suggest that salt-bridge formation between residues located on the same face of a transmembrane helix may reduce the free energy of membrane partitioning. PMID- 11021966 TI - Epstein-barr virus nuclear antigen 2 retards cell growth, induces p21(WAF1) expression, and modulates p53 activity post-translationally. AB - The Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA2) has been shown to be required for promotion of cell-cycle progression in EBV-immortalized B lymphocytes. However, other studies have indicated that EBNA2 alone, in the absence of other EBV genes, may retard cell growth. To resolve this discrepancy, we investigated the effect of EBNA2 on the growth of various cells, including EBV target nasopharyngeal carcinoma cells, NPC-TW01 and NPC-TW04. We found that EBNA2 could retard cell growth, in stable Vero, HEp-2, and U2OS cell clones expressing EBNA2, and in Vero, 293, NPC-TW01, and NPC-TW04 cells transiently transfected with EBNA2. While investigating the mechanism underlying the growth-retarding function of EBNA2, we found that EBNA2 induced p21(WAF1) expression in these cells. This induction of p21(WAF1) expression was mediated through p53. EBNA2 was found to stimulate p53 to bind to the p53-response element within the p21(WAF1) promoter, possibly by promoting p53 phosphorylation. This enhancement of p53 sequence-specific DNA-binding activity may be the mechanism through which EBNA2 activates the expression of p53-regulated genes, including p21(WAF1) and mdm-2. Together, these studies reveal a possible intrinsic function of EBNA2 in cell growth regulation and elucidate a novel mechanism by which EBNA2 modulates transcription. PMID- 11021967 TI - Archaeal histone selection of nucleosome positioning sequences and the procaryotic origin of histone-dependent genome evolution. AB - Archaeal histones and the eucaryal (eucaryotic) nucleosome core histones have almost identical histone folds. Here, we show that DNA molecules selectively incorporated by rHMfB (recombinant archaeal histone B from Methanothermus fervidus) into archaeal nucleosomes from a mixture of approximately 10(14) random sequence molecules contain sequence motifs shown previously to direct eucaryal nucleosome positioning. The dinucleotides GC, AA (=TT) and TA are repeated at approximately 10 bp intervals, with the GC harmonic displaced approximately 5 bp from the AA and TA harmonics [(GCN(3)AA or TA)(n)]. AT and CG were not strongly selected, indicating that TA not equalAT and GC not equalCG in terms of facilitating archaeal nucleosome assembly. The selected molecules have affinities for rHMfB ranging from approximately 9 to 18-fold higher than the level of affinity of the starting population, and direct the positioned assembly of archaeal nucleosomes. Fourier-transform analyses have revealed that AA dinucleotides are much enriched at approximately 10. 1 bp intervals, the helical repeat of DNA wrapped around a nucleosome, in the genomes of Eucarya and the histone-containing Euryarchaeota, but not in the genomes of Bacteria and Crenarchaeota, procaryotes that do not have histones. Facilitating histone packaging of genomic DNA has apparently therefore imposed constraints on genome sequence evolution, and since archaeal histones have no structure in addition to the histone fold, these constraints must result predominantly from histone fold DNA contacts. Based on the three-domain universal phylogeny, histones and histone dependent genome sequence evolution most likely evolved after the bacterial archaeal divergence but before the archaeal-eucaryal divergence, and were subsequently lost in the Crenarchaeota. However, with lateral gene transfer, the first histone fold could alternatively have evolved after the archaeal-eucaryal divergence, early in either the euryarchaeal or eucaryal lineages. PMID- 11021968 TI - Crystal structures of recombinant histones HMfA and HMfB from the hyperthermophilic archaeon Methanothermus fervidus. AB - The hyperthermophilic archaeon Methanothermus fervidus contains two small basic proteins, HMfA (68 amino acid residues) and HMfB (69 residues) that share a common ancestry with the eukaryal nucleosome core histones H2A, H2B, H3, and H4. HMfA and HMfB have sequences that differ at 11 locations, they have different structural stabilities, and the complexes that they form with DNA have different electrophoretic mobilities. Here, crystal structures are documented for recombinant (r) HMfA at a resolution of 1.55 A refined to a crystallographic R value of 19.8 % (tetragonal form) and at 1.48 A refined to a R-value of 18.8 % (orthorhombic form), and for rHMfB at 1.9 A refined to a R-value of 18.0 %. The rHMfA and rHMfB monomers have structures that are just histone folds in which a long central alpha-helix (alpha2; 29 residues) is separated from shorter N terminal (alpha1; 11 residues) and C-terminal (alpha3; 10 residues) alpha-helices by two loops (L1 and L2; both 6 residues). Within L1 and L2, three adjacent residues are in extended (beta) conformation. rHMfA and rHMfB assemble into homodimers, with the alpha2 helices anti-parallel aligned and crossing at an angle of close to 35 degrees, and with hydrogen bonds formed between the extended, parallel regions of L1 and L2 resulting in short beta-ladders. Dimerization creates a novel N-terminal structure that contains four proline residues, two from each monomer. As prolines are present at these positions in all archaeal histone sequences, this proline-tetrad structure is likely to be a common feature of all archaeal histone dimers. Almost all residues that participate in monomer-monomer interactions are conserved in HMfA and HMfB, consistent with the ability of these monomers to form both homodimers and (HMfA+HMfB) heterodimers. Differences in side-chain interactions that result from non-conservative residue differences in HMfA and HMfB are identified, and the structure of a (rHMfA)(2)-DNA complex is presented based on the structures documented here and modeled by homology to histone-DNA interactions in the eukaryal nucleosome. PMID- 11021970 TI - Analysis and prediction of functional sub-types from protein sequence alignments. AB - The increasing number and diversity of protein sequence families requires new methods to define and predict details regarding function. Here, we present a method for analysis and prediction of functional sub-types from multiple protein sequence alignments. Given an alignment and set of proteins grouped into sub types according to some definition of function, such as enzymatic specificity, the method identifies positions that are indicative of functional differences by comparison of sub-type specific sequence profiles, and analysis of positional entropy in the alignment. Alignment positions with significantly high positional relative entropy correlate with those known to be involved in defining sub-types for nucleotidyl cyclases, protein kinases, lactate/malate dehydrogenases and trypsin-like serine proteases. We highlight new positions for these proteins that suggest additional experiments to elucidate the basis of specificity. The method is also able to predict sub-type for unclassified sequences. We assess several variations on a prediction method, and compare them to simple sequence comparisons. For assessment, we remove close homologues to the sequence for which a prediction is to be made (by a sequence identity above a threshold). This simulates situations where a protein is known to belong to a protein family, but is not a close relative of another protein of known sub-type. Considering the four families above, and a sequence identity threshold of 30 %, our best method gives an accuracy of 96 % compared to 80 % obtained for sequence similarity and 74 % for BLAST. We describe the derivation of a set of sub-type groupings derived from an automated parsing of alignments from PFAM and the SWISSPROT database, and use this to perform a large-scale assessment. The best method gives an average accuracy of 94 % compared to 68 % for sequence similarity and 79 % for BLAST. We discuss implications for experimental design, genome annotation and the prediction of protein function and protein intra-residue distances. PMID- 11021969 TI - Three-dimensional structure of a human pancreatic ribonuclease variant, a step forward in the design of cytotoxic ribonucleases. AB - We have determined the crystal structure of a human pancreatic ribonuclease or RNase 1 variant at 1.65 A resolution. Five residues in the N-terminal region were substituted by the corresponding amino acids of the bovine seminal RNase. In addition, a Pro to Ser mutation was present at position 50. The substitution of part of the N terminus has been critical both in improving the expression of this enzyme as a recombinant protein and in achieving its crystallisation. The determination of the crystal structure revealed the characteristic RNase fold including a V-shaped beta-sheet and three alpha-helices. It differs from its bovine RNase orthologue mainly in the loop regions. The active-site cleft shows a similar architecture to that of its bovine counterpart, with the essential residues occupying equivalent positions. In the present structure, however, His119 is displaced as it is in the structure of RNase A at high pH. An interaction model of human ribonuclease with the ribonuclease inhibitor, together with inhibition assays, indicate that, in contrast to RNase A, the modification of the loop beta4beta5 is not enough to avoid inhibition. This study represents the first crystallographic approach to the human enzyme, and should constitute an invaluable tool for the design of ribonuclease variants with acquired cytotoxic properties. PMID- 11021971 TI - Probing the catalytic mechanism of GDP-4-keto-6-deoxy-d-mannose Epimerase/Reductase by kinetic and crystallographic characterization of site specific mutants. AB - GDP-4-keto-6-deoxy-d-mannose epimerase/reductase is a bifunctional enzyme responsible for the last step in the biosynthesis of GDP-l-fucose, the substrate of fucosyl transferases. Several cell-surface antigens, including the leukocyte Lewis system and cell-surface antigens in pathogenic bacteria, depend on the availability of GDP-l-fucose for their expression. Therefore, the enzyme is a potential target for therapy in pathological states depending on selectin mediated cell-to-cell interactions. Previous crystallographic investigations have shown that GDP-4-keto-6-deoxy-d-mannose epimerase/reductase belongs to the short chain dehydrogenase/reductase protein homology family. The enzyme active-site region is at the interface of an N-terminal NADPH-binding domain and a C-terminal domain, held to bind the substrate. The design, expression and functional characterization of seven site-specific mutant forms of GDP-4-keto-6-deoxy-d mannose epimerase/reductase are reported here. In parallel, the crystal structures of the native holoenzyme and of three mutants (Ser107Ala, Tyr136Glu and Lys140Arg) have been investigated and refined at 1. 45-1.60 A resolution, based on synchrotron data (R-factors range between 12.6 % and 13.9 %). The refined protein models show that besides the active-site residues Ser107, Tyr136 and Lys140, whose mutations impair the overall enzymatic activity and may affect the coenzyme binding mode, side-chains capable of proton exchange, located around the expected substrate (GDP-4-keto-6-deoxy-d-mannose) binding pocket, are selectively required during the epimerization and reduction steps. Among these, Cys109 and His179 may play a primary role in proton exchange between the enzyme and the epimerization catalytic intermediates. Finally, the additional role of mutated active-site residues involved in substrate recognition and in enzyme stability has been analyzed. PMID- 11021972 TI - Molecular enzymology of the EcoRV DNA-(Adenine-N (6))-methyltransferase: kinetics of DNA binding and bending, kinetic mechanism and linear diffusion of the enzyme on DNA. AB - The EcoRV DNA-(adenine-N(6))-methyltransferase recognizes GATATC sequences and modifies the first adenine residue within this site. We show here, that the enzyme binds to the DNA and the cofactor S-adenosylmethionine (AdoMet) in an ordered bi-bi fashion, with AdoMet being bound first. M.EcoRV binds DNA in a non specific manner and the enzyme searches for its recognition site by linear diffusion with a range of approximately 1800 bp. During linear diffusion the enzyme continuously scans the DNA for the presence of recognition sites. Upon specific M.EcoRV-DNA complex formation a strong increase in the fluorescence of an oligonucleotide containing a 2-aminopurine base analogue at the GAT-2AP-TC position is observed which, most likely, is correlated with DNA bending. In contrast to the GAT-2AP-TC substrate, a G-2AP-TATC substrate in which the target base is replaced by 2-aminopurine does not show an increase in fluorescence upon M.EcoRV binding, demonstrating that 2-aminopurine is not a general tool to detect base flipping. Stopped-flow experiments show that DNA bending is a fast process with rate constants >10 s(-1). In the presence of cofactor, the specific complex adopts a second conformation, in which the target sequence is more tightly contacted by the enzyme. M.EcoRV exists in an open and in a closed state that are in slow equilibrium. Closing the open state is a slow process (rate constant approximately 0.7 min(-1)) that limits the rate of DNA methylation under single turnover conditions. Product release requires opening of the closed complex which is very slow (rate constant approximately 0.05-0.1 min(-1)) and limits the rate of DNA methylation under multiple turnover conditions. M.EcoRV methylates DNA sequences containing more than one recognition sites in a distributive manner. Since the dissociation rate from non-specific DNA does not depend on the length of the DNA fragment, DNA dissociation does not preferentially occur at the ends of the DNA. PMID- 11021973 TI - Galanin and spinal nociceptive mechanisms: recent advances and therapeutic implications. AB - Galanin is a peptide consisting of 29 or 30 (in humans) amino acids that is present in sensory and spinal dorsal horn neurons. Endogenous galanin may have an important modulatory function on nociceptive input at the spinal level. In addition, exogenously administered galanin exerts complex effects on spinal nociceptive transmission, where inhibitory action appears to predominate. Peripheral nerve injury and inflammation, conditions associated with chronic pain, upregulate the synthesis of galanin in sensory neurons and spinal cord neurons, respectively. Hence, the sensory effect of galanin may be increased under these conditions, raising the possibility that modulation of the activity of the galanin system may produce antinociception. PMID- 11021974 TI - Conditioned fear-induced changes in neuropeptide Y-like immunoreactivity in rats: the effect of diazepam and buspirone. AB - The study aimed to investigate the influence of conditioned fear, produced in the passive avoidance test, on neuropeptide Y-like immunoreactivity (NPY-LI) and the effect of anxiolytics on NPY-LI in frightened rats. Rats avoided the dark chamber, where they were previously subjected to electric footshock, and they exhibited increased numbers of defecations and gastric ulcers. Moreover, they showed increased NPY-LI in the amygdala, nucleus accumbens and hypothalamus, and decreased NPY-LI in the frontal cortex. Diazepam (1 or 3 mg/kg) and buspirone (1.5 or 5 mg/kg) dose-dependently inhibited passive avoidance and decreased the numbers of defecations, and they also decreased the number of gastric ulcers. Diazepam reversed while buspirone only attenuated the fear-induced changes in NPY LI in all regions studied. In the amygdala, the effect of diazepam was dose dependent. The effect of diazepam on both behaviour and NPY-LI was antagonized by flumazenil (15 mg/kg). Apart from supporting the role of the NPY system in fear and anxiety, the results of this study suggest that NPY is involved in the anxiolytic effects of diazepam and buspirone and that the effect of diazepam is mediated by benzodiazepine receptors. PMID- 11021975 TI - Supraspinal NMDA and non-NMDA receptors are differentially involved in the production of antinociception by morphine and beta-endorphin administered intracerebroventricularly in the formalin pain model. AB - Our previous studies have demonstrated that supraspinal glutamate receptors are differentially involved in the antinociception induced by morphine and beta endorphin given intracerebroventricularly (i.c.v.) in the tail-flick and hot plate tests. The formalin pain test was used in the present study. Injection of mice with formalin solution (2%, 10 microl) into the hindpaw intraplantarly produced the first (0-5 min) and second (20-40 min) phases of formalin responses. The formalin responses in the both phases were attenuated dose-dependently by morphine (0.125-1 microg) or beta-endorphin (0.125-1 microg) administered i.c.v. 5 min before. The antinociceptive effect of morphine was slightly more potent in the second phase whereas the effect of beta-endorphin was more pronounced in the first phase. MK-801 (0.1-1 microg), a non-competitive NMDA receptor antagonist, and CNQX (0.05-0.5 microg), a non-NMDA antagonist, given i.c.v., produced antinociceptive effect in the both phases, but only in a partial manner. Both MK 801 (0.05 microg) and CNQX (0.01 microg), at the dose which had no intrinsic effect, reversed the antinociceptive effect of beta-endorphin (1 microg) observed during the second, but not the first, phase partially but significantly. However, the antinociceptive effect of morphine (1 microg) was not affected by the same dose of MK-801 or CNQX given i.c.v. Our results indicate that, at the supraspinal level, both NMDA and non-NMDA receptors are involved in the production of antinociception induced by supraspinally administered beta-endorphin, but not morphine, in the formalin pain model. PMID- 11021977 TI - Vasopressin and oxytocin receptor mRNA expression during rat telencephalon development. AB - We investigated the developmental expression of vasopressin and oxytocin receptor and peptide mRNA using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Messenger RNAs for both vasopressin receptor subtypes V(1)a and V(2)were present in the telencephalon from embryonic day 12 to day 20. Both V(1)a and V(2)receptor mRNA increased on day 13 and then remained stable from embryonic day 13 to day 20. Messenger RNA for the vasopressin peptide was also detected in the telencephalon from day 12 to day 20, indicating that vasopressin could be synthesized within the rat cerebral cortex during rat embryonic development. Oxytocin receptor mRNA expression was also present in the telencephalon, but expression levels varied considerably from day 12 to day 20. No oxytocin mRNA expression was detected during rat telencephalon development. Temporal patterns of vasopressin receptor and vasopressin peptide mRNA expression along with oxytocin receptor mRNA suggest a temporal role for vasopressin- and oxytocin-mediated actions during rat telencephalon development. PMID- 11021976 TI - Antagonism of NPY-induced feeding by pretreatment with cyclic AMP response element binding protein antisense oligonucleotide. AB - Although second messenger systems subserving neuropeptide Y (NPY)-mediated behaviors have been identified for a variety of receptors in several tissues, downstream signaling events are not well known. The nuclear binding protein, cyclic AMP response element binding protein (CREB) appears to be a transcription factor that is activated following injection of NPY into rat hypothalamus. To allow determination of the functional nature of CREB mediation of NPY-induced feeding, injection cannulae were implanted into the perifornical hypothalamus of 18 rats. Treatment of seven rats with CREB antisense oligonucleotide (15 ug) significantly antagonized NPY feeding for up to one week after treatment, while similar injections of CREB sense oligonucleotide (15 ug) had no significant effect on NPY-induced feeding. Two weeks after the antisense oligonucleotide treatment, feeding was once again elicited by the injection of NPY. Hypothalamic CREB protein was also reduced significantly two days after the CREB antisense oligonucleotide treatment. These results suggest that activation of CREB, probably through phosphorylation, may be a necessary event for the signal transduction of NPY stimulation into feeding behavior. PMID- 11021978 TI - Effect of opioid active therapeutics on the ascending reflex pathway in the rat ileum. AB - For a long time therapeutic agents that interact with opioid receptors have been used in antidiarrheal therapy. The action of the opioid active substances on motility and transit have already been characterized; however, their effects on myenteric reflexes and their possible luminal action have not yet been investigated. Loperamide, fedotozine and beta-casomorphin-4, as well as the casomorphin-analogue beta-CM-4027, are, or have been, suggested as therapeutic agents and were studied in the isolated rat ileum for their effect on the ascending reflex pathway. beta-CM-4027 > fedotozine > loperamide > beta casomorphin-4 caused a concentration-dependent inhibition of the ascending contractile reflex response with an IC(50)of 1.4x10(-7)M, 1.5x10(-6)M, 4.1x10( 6)M and 4.5x10(-6)M respectively. At the same time as the oral contractile reflex response was inhibited, all four opioid agonists (CM-4027 > beta-casomorphin-4 > fedotozine > loperamide) increased the latency of the reflex response. Both effects were blocked by naloxone, indicating the involvement of opioid receptors. These results demonstrate that opioid-active drugs and substances modify the peristaltic reflex by reducing the efficacy of the reflex response and modulating the timing of the reflex pathway. In a second series of experiments, luminal application of opioid-active drugs was compared with serosal application. beta casomorphine-4 caused a concentration-dependent inhibition of the oral reflex response with an IC(50)of 3x10(-3)M which was 750 times higher than after serosal application. In contrast, a stable and highly selective kappa opioid agonist (U 50,488), which caused potent inhibition upon serosal application (IC(50): 2.3x10( 7)M), showed no inhibitory effect after luminal application up to a concentration of 10(-2)M. Thus casomorphins could have a local effect on the gut wall with no need for systemic absorption. This might be used for a possible therapeutic application. PMID- 11021979 TI - Augmenting effect of methionine-enkephalin on interleukin-6 production by cytokine-stimulated murine macrophages. AB - The effects of methionine-enkephalin on the production of interleukin-6 by activated peritoneal murine macrophages were studied. Macrophage were activated with interleukin-1beta or interferon-gamma in the presence or absence of graded concentrations of methionine-enkephalin. Methionine-enkephalin combined with interleukin-1beta or interferon-gamma caused an increase in IL-6 release from cultured macrophages. The opioid receptor antagonist naloxone did not change the stimulatory effect of methionine-enkephalin on IL-6 production by stimulated macrophages. Methionine-enkephalin added to the culture medium of resting macrophages increased IL-6 release from macrophages which were later induced with interleukin-1beta or interferon-gamma. The results of this study suggest that methionine-enkephalin can modulate the proinflammatory cytokine response by controlling, via non-opioid receptor mechanism, the production of IL-6. PMID- 11021980 TI - Effects of the peripherally selective kappa-opioid asimadoline, on substance P and CGRP mRNA expression in chronic arthritis of the rat. AB - We have previously shown that the kappa-opioid agonist, asimadoline, produces time-dependent changes in neuropeptide concentrations in the joints of rats with chronic arthritis. We hypothesized that asimadoline acts on peripheral terminals to modulate substance P (SP) release. To address this hypothesis, here we have examined neuropeptide expression in their source cells in dorsal root ganglia (DRG) that innervate the joint, as well as in non-neuronal tissue, after treatment with asimadoline. We found an increased production of SP and CGRP in untreated chronic arthritic animals which supports our previous finding of increased SP content in the joint. More importantly, the kappa-opioid asimadoline reduced the expression of both SP and calcitonin gene-related peptide-alpha (alpha-CGRP) in DRG cells but had no effect on the very low expression of neuropeptides in non-neuronal tissue. The fact that SP synthesis is attenuated by asimadoline accords with our hypothesis that the increased tissue levels of SP result from kappa-mediated pre-synaptic inhibition of release leading to augmented tissue stores. These in vivo data confirm literature findings that opioids inhibit SP release from peripheral endings of primary afferent fibres. PMID- 11021981 TI - Effects of angiotensin II on visual evoked potentials in the superior colliculus of juvenile rats. AB - There are age-related changes in the relative expression of the AT(1)and AT(2)receptors of angiotensin II (Ang II) in brain regions such as the superior colliculus, a midbrain visual structure where both receptor subtypes are found. We investigated the effects of Ang II on gross visual activity in the colliculus of anesthetized rats aged between 15 and 35 post-natal days. Microinjection of Ang II in the superficial layers yielded a strong reduction in the amplitude of visual evoked potentials in a dose-related manner. Injection of the peptide in more ventral collicular layers did not modify the potential confirming the discrete localization of the angiotensinergic receptors in the superficial layers. Preliminary data indicated that the co-injection of Ang II with Losartan or PD 123319 yielded a partial blockade of Ang II suppressive effects, indicating that both AT(1)and AT(2)receptors are likely to be involved in mediating these responses. Overall, this study shows that the inhibitory nature of Ang II action is similar in juvenile and adult animals (Merabet et al. 1994 and Merabet et al. 1997) PMID- 11021982 TI - The role of melanocortin receptors in sexual behavior in female rats. AB - Earlier data have indicated that alpha-MSH may play a role for sexual behavior in rats. In this study we investigated the effects of MSH peptides on sexual receptivity in ovariectomized-adrenalectomized female rats, pre-treated with benzoate of estradiol, in presence of vigorous male rats. The results show that alpha-MSH significantly increases lordosis behavior in female rats after injections into the ventromedial nucleus. Interestingly, we have for the first time shown that gamma-MSH also causes significant increase in lordosis in female rats. Furthermore, we show that HS014, an antagonist for the central MC receptors, in dose dependent manner blocks the effect of alpha-MSH on lordosis. The results indicate that the effects of MSH peptides on female sexual behaviour are mediated through a specific MC receptor, which could be the MC3 receptor. PMID- 11021983 TI - Inhibition of nociceptive withdrawal reflex by microinjection of interleukin 2 into rat locus coeruleus. AB - This study was to examine the effects of microinjection of human recombinant interleukin 2 (IL-2) into locus coeruleus (LC) on spinal nociception. Following application of IL-2 (0.1 microl, 10 pM) into LC, the percentage of inhibition of nociceptive C responses of reflex at 3, 9, 15, 21 and 27 min after injection were 88.2 +/-9.4%, 84.0 +/- 11.8%, 89.7 +/- 10.5%, 57.1 +/- 8.7% and 26.3 +/- 12.2%, respectively. Also, the expression of Fos protein in superficial dorsal horn was reduced by 73.01 +/- 13.58% of control (P<0.0001). Naloxone (10 microg, i.p.) completely blocked the IL-2-induced inhibition of C responses. The results clearly show that IL-2 receptors present in LC mediate descending inhibition of the spinal nociception, which may couple with the activation of opioid receptors on LC neurons. PMID- 11021984 TI - Hippocampal gene expression in the pig: upregulation of corticotrophin releasing hormone mRNA following central administration of the peptide. AB - Corticotrophin releasing hormone (CRH) and glucocorticoids affect hypophysiotrophic regions of the brain and influence limbic system activity. Since the latter mediates emotional responses, changes in gene expression in regions such as the hippocampus may provide new information on neural stress mechanisms. In this study, mRNA for CRH and selected ionotropic glutamate receptor (iGluR) subunits (NR1, GluR2, GluR3) was quantified in the hippocampus of pigs in which stress was simulated by central administration of CRH (100 microg). Increases in hippocampal CRH mRNA were detected in the CA3 subfield 4 h later, and in the CA1, CA2 and CA3 subfields 24 h post-treatment. However, there were no associated changes in iGluR subunit mRNAs, although the ratio GluR3: GluR2 increased in the dentate gyrus after 4 h. These results, together with a recent similar finding in rats subjected to restraint, point to an involvement of hippocampal CRH in the neuronal response to stress. PMID- 11021985 TI - Involvement of cAMP-dependent protein kinase and pertussis toxin-sensitive G proteins in CGRP mediated JNK activation in human neuroblastoma cell line. AB - Calcitonin gene-related peptide (CGRP) is a neuropeptide with potent cardiovascular effects, which include positive inotropic and chronotropic actions, systemic vasodilation, and hypotension in animal and human studies. Human neuroblastoma cells (SK-N-MC) have been used as a model system to study the CGRP receptors and downstream signaling pathways. This investigation was undertaken to study the role of CGRP in the activation of mitogen-activated protein kinases. While exposure of these cells to CGRP had no significant effect on ERK-1 or p38 MAP kinases, JNK activity was stimulated by CGRP in a time- and concentration-dependent fashion. CGRP-mediated JNK-activation was inhibited by CGRP receptor antagonist, CGRP8-37, confirming that this is a receptor-mediated event. In addition, pretreatment of the cells with H-89, protein kinase A inhibitor or pertussis toxin greatly attenuated CGRP-mediated JNK activation suggesting the requirement of cAMP-dependent protein kinase activation and involvement of pertussis toxin-sensitive G-protein in CGRP-mediated JNK activation. PMID- 11021986 TI - Effect of corticotropin releasing factor receptor 1 antagonist on extracellular norepinephrine, dopamine and serotonin in hippocampus and prefrontal cortex of rats in vivo. AB - Corticotropin releasing factor (CRF) is a major mediator of adaptive responsiveness to stress. We measured changes in extracellular concentrations of catecholamine and indoleamines in freely moving rats in response to administration of CRF1 antagonist CP-154,526 by using in vivo microdialysis. Dialysis probes were placed stereotaxically in either the hippocampus or the prefrontal cortex. We examined the response in the hippocampus or the prefrontal cortex to 32.0 mg/kg i.p. administration of CP-154,526. CP-154,526 reduced the extracellular concentration of norepinephrine (NE) from 30 min to 180 min and 5 hydroxytryptamine (5-HT) from 30 min to 60 min after injection in the hippocampus. CP-154,526 did not remarkably change dopamine (DA). There were no significant differences between CP-154,526 and vehicle in NE, 5-HT and DA in the prefrontal cortex. The present results indicate that CRF1 receptor antagonist produced a decrease in dialysate concentration of NE and 5-HT, but not DA, in the hippocampus. These results suggest that the CRH-1 receptor antagonist suppresses the release of NE and 5-HT in the hippocampus. PMID- 11021988 TI - Exploitation of cellular signaling by HIV-1: unwelcome guests with master keys that signal their entry. PMID- 11021987 TI - Characterization of [Phe(13), Tyr(19)]-MCH analog binding activity to the MCH receptor. AB - Melanin concentrating hormone (MCH), a hypothalamic neuropeptide, is an important regulator of energy homeostasis in mammals. Characterization of an MCH specific receptor has been hampered by the lack of a suitable radioligand. The [Phe(13), Tyr(19)]-MCH analog has been shown by different investigators to bind specifically to cell lines of epithelial or pigment cell origin. Recently, using functional assays, the MCH receptor has been characterized as a seven transmembrane G-coupled protein initially identified as SLC-1. In the present study, we used tyrosine iodinated [Phe(13), Tyr(19)]-MCH analog, which stimulates food intake in a manner similar to that of MCH, as well as native MCH to conduct binding studies. Specific binding could not be demonstrated in intact cells of several cell lines, including A431 and B16. Specific binding associated with membranes localized to the microsomal, not the plasma membrane, fraction. Message for SLC-1 was absent in these cell lines, as assessed by Northern blot analysis. We conclude that cells previously reported to express the MCH receptor do not express SLC-1 and that both iodinated MCH and the [Phe(13), Tyr(19)]-MCH have a large component of non-specific binding. These ligands may be useful for binding studies in transfected cells with high levels of SLC-1 expression. However they do not appear to be suitable for screening for the MCH receptor as most cells demonstrate significant low affinity non-specific binding. PMID- 11021989 TI - In vitro strand transfer from broken RNAs results in mismatch but not frameshift mutations. AB - An in vitro system to compare the fidelity of strand transfers from truncated vs full-length RNAs was constructed. A donor RNA, on which reverse transcriptase (RT)-directed DNA synthesis was initiated, shared homology with an acceptor RNA, to which DNAs initiated on the donor could transfer. All RNAs were derived from the N-terminal portion of the alpha-lac gene. On full-length donors, transfers occurred when DNAs migrated to the acceptor prior to being completed on the donor. On donors that were truncated, most transfers occurred after DNAs reached the end of the donor. Transfer products were amplified by PCR and used to replace the corresponding region in a vector containing the alpha-lac gene. Transformed Escherichia coli were screened for alpha-complementation by blue-white phenotype analysis, with white colonies scored as those with errors in alpha-lac. These errors were derived from RT synthesis and strand transfer. The mutant colony frequency approximately doubled for transfer products derived from truncated donors (0.026+/-0.005 vs. 0.053+/-0.011 (three experiments +/- SD), for full length vs. truncated, respectively). The increases resulted from additional non template-directed bases (mostly thymidines) added to the DNAs before transfer. Sequence analysis of DNAs synthesized on truncated donors showed that about 60% had additions (20/34); however, those without additions transferred at a much higher rate than those with. Transfer of the DNAs with additions always resulted in substitutions; no frameshifts were observed. Results are consistent with RT adding nontemplated nucleotides at template termini. Transfer and subsequent extension of these products is severely inhibited relative to products without additions. The potential relevance of these findings to retrovirus replication is discussed. PMID- 11021990 TI - Induction of apoptosis by the Vpr protein of human immunodeficiency virus type 1 occurs independently of G(2) arrest of the cell cycle. AB - The HIV-1 accessory gene product Vpr can inhibit cell proliferation via cell cycle arrest at the G(2) phase, and it can induce apoptosis after G(2) arrest. We found recently that C81, a carboxy-terminally truncated form of Vpr, induced apoptosis via G(1) arrest but did not induce G(2) arrest of the cell cycle. Thus, it seemed possible that expression of Vpr in cells might cause apoptosis independently of the ability of Vpr to induce G(2) arrest. We demonstrate here that Vpr-induced apoptosis occurs by a mechanism that does not necessarily require induction of G(2) arrest. First, it was found that the extent of apoptosis reached a maximum even when few cells were arrested at the G(2) phase of the cell cycle and was reduced in inverse proportion to the increased induction of G(2) arrest. Thus, the extent of induction of G(2) arrest was not correlated with the extent of Vpr-induced apoptosis. Furthermore, we replaced the Ile/Leu residues in the leucine zipper-like domain of Vpr with Ala or Pro and used cells that expressed the mutant protein to demonstrate that Vpr caused apoptosis in a manner that was independent of G(2) arrest. Finally, replacement of Ile/Leu by Pro at positions 60, 67, 74, and 81 within the leucine zipper-like domain of wild-type Vpr and C81 revealed that the Ile/Leu residues at positions 60, 67, and 74 in the leucine zipper-like domain were indispensable for induction of apoptosis induced by Vpr and by C81 and confirmed, in addition, that both processes might be regulated by the same pathway. C81 appears to be a useful tool for elucidation of the mechanism of apoptosis induced by expression of Vpr protein. PMID- 11021991 TI - Characterization of a beta-1,3-glucanase encoded by chlorella virus PBCV-1. AB - Sequence analysis of the 330-kb chlorella virus PBCV-1 genome revealed an open reading frame, A94L, that encodes a protein with significant amino acid identity to Glycoside Hydrolase Family 16 beta-1,3-glucanases. The a94l gene was cloned and the protein was expressed as a GST-A94L fusion protein in Escherichia coli. The recombinant A94L protein hydrolyzed the beta-1,3-glucose polymer laminarin and had slightly less hydrolytic activity on beta-1,3-1, 4-glucose polymers, lichenan and barley beta-glucan. The recombinant enzyme had the highest activity at 65 degrees C and pH 8. We predicted that the a94l-encoded beta-1,3-glucanase is involved in degrading the host cell wall either during virus release and/or is packaged in the virion particle and involved in virus entry. Therefore, we expected a94l to be expressed late in virus infection. However, contrary to expectations, both the a94l mRNA and the A94L protein appeared 15 min after PBCV 1 infection and disappeared 60- and 120-min p.i. postinfection, respectively, indicating that a94l is an early gene. Twenty-seven of 42 chlorella viruses contained the a94l gene. To our knowledge, this is the first report of a virus encoded beta-1,3-glucanase. PMID- 11021992 TI - Identification and characterization of recombinant subgroup J avian leukosis viruses (ALV) expressing subgroup A ALV envelope. AB - Three recombinant avian leukosis subgroup J viruses, ADOL 5701A, ADOL 5701ADelta, and ADOL 6803A, carrying a subgroup A envelope have been isolated and characterized. These viruses were identified by their ability to replicate in DF 1/J, a recombinant chicken embryo fibroblast (CEF) cell line expressing the subgroup J envelope that is resistant to subgroup J replication. Flow cytometric analysis of DF-1/J cells infected with ADOL 5701 and ADOL 6803, two subgroup J isolates, indicated cross-reactivity with subgroup A chicken polyclonal serum. Based on published sequences of subgroups A and J isolates, we designed a series of primers to PCR amplify the envelope and LTR of these viruses. PCR products were obtained when the forward primer was specific for subgroup A gp85 envelope protein gene and the reverse primer was specific for subgroup J LTR. Sequence analysis of the PCR products indicated that these viruses had a subgroup A gp85, a subgroup E gp37, and a subgroup J LTR. Interestingly, these viruses had previously been propagated in CEF from the alv6 chicken line, a line that carries a replication defective recombinant endogenous virus expressing a subgroup A envelope (RAV 0-A(1)). Sequence analysis of RAV 0-A(1) gp85 and gp37 envelope proteins indicated that they were almost identical to those of the recombinants ADOL 5701A and ADOL 6803A. These results indicate that these three recombinant viruses arose by recombination between exogenous subgroup J isolates and a recombinant defective endogenous virus with subgroup A envelope. PMID- 11021993 TI - NSAID treatment suppresses VSV propagation in mouse CNS. AB - Cyclooxygenase (COX) is the key enzyme in the conversion of arachidonic acid to prostaglandins. COX has two isoforms: COX-1, the constitutively expressed form, and COX-2, the inducible form. Prostaglandins are mediators of many critical physiological and inflammatory responses, but little is known about their roles during a viral infection in the central nervous system (CNS). We used non selective inhibitors of COX, aspirin and indomethacin, and a selective antagonist of COX-2, celecoxib, to study the role of prostaglandins in Vesicular Stomatitis Virus (VSV) induced encephalitis. We found that the inhibition of COX antagonizes VSV propagation both in vitro and in vivo. In addition, aspirin and celecoxib both prevented the disruption of the blood brain barrier in VSV-infected mice. In vitro experiments showed that the effect of COX inhibition was at least partially mediated by increased production of Nitric Oxide (NO), a molecule known to inhibit VSV replication. When NO production was inhibited by N(omega)-nitro-L methyl-arginine-ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, the difference in viral titer between aspirin (or celecoxib)-treated and the control cells was abolished. VSV-infected mice treated with celecoxib expressed more NOS 1 and produced more NO in their CNS compared to the controls. Our data suggest that the product(s) of COX have antagonistic effect(s) on NO production in the mouse CNS. PMID- 11021994 TI - Epstein-Barr virus lacking glycoprotein gp85 cannot infect B cells and epithelial cells. AB - To evaluate whether the BXLF2 gene of Epstein-Barr virus (EBV), which codes gp85 protein, is essential for infection of B cells and epithelial cells, we analyzed the infectivity of an EBV recombinant lacking gp85. The cells that were infected with the BXLF2-disrupted virus were unable to express gp85 proteins that could be detected by mouse monoclonal antibody E1D1, specific for gp85/gp25 complexes. The BXLF2-disrupted EBV had the ability to attach to, but not infect, B cells. On the other hand, the same virus failed to bind to and infect NU-GC-3, a human gastric adenocarcinoma cell line that is susceptible to EBV infection. The results indicate that the gp85 is used for infection of not only B cells but also epithelial cells and suggest that the gp85 is necessary for attaching the virus to epithelial cells. PMID- 11021995 TI - Distribution and quantification of human immunodeficiency virus type 1, strain JC499, proviral DNA in tissues from an infected chimpanzee. AB - Data are accumulating to show that the natural history of human immunodeficiency virus type 1 (HIV-1) in chimpanzees closely reproduces that in humans and is influenced by biologic properties of the infecting HIV-1 strain. To determine the distribution and relative amounts of HIV-1, proviral DNA in multiple tissues from a chimpanzee euthanized because of an abdominal tumor and kidney failure was quantified by nested PCR limiting-dilution assays. At death, 21 months after infection with HIV-1(JC499), this animal had a CD4(+) T-cell count of 268 and 1.7 x 10(5) copies of virion RNA/ml of plasma. The highest proviral burdens were in peripheral lymph nodes and blood, followed by lung, colon, and spleen; values ranged from 130 to 3350 proviral copies/microg DNA (equivalent to DNA in 150,000 cells). The lowest levels of virus were in the spinal cord, brain, and cecum (0.3 to 2.5 copies/microg DNA), with all other tissues harboring intermediate levels (6.8 to 114 copies/microg DNA). Viral burdens in all tissues were comparable to or greater than those reported for HIV-1-infected humans in all stages of disease. Immunohistochemistry for HIV-1 p24 Gag antigen revealed (i) trapping of HIV-1 on follicular dendritic cells in lymph node germinal centers and (ii) virus in the brain, where it was localized primarily to capillary endothelial cells in the cerebral cortex. Analysis of the genetic diversity of the Env V3 loop in tissues indicated that there was no apparent compartmentalization of HIV-1 variants. Of interest, in 83 of 94 (88.3%) clones sequenced, the unique GYGR motif at the tip of the V3 loop of HIV-1(JC499) had reverted to the more common GPGR. The results support the conclusion that HIV-1 has the potential to maintain high viral burdens in chimpanzees and to disseminate to most organs, including the central nervous system. The use of the chimpanzee model with HIV-1(JC499) (or related strains) in vaccine efficacy studies should prove valuable, especially when assessing protection against disease. Furthermore, comparison of both replicative properties of HIV-1(JC499) with SIVcpz strains and immune responses of chimpanzees infected with these viruses might provide new information about HIV pathogenesis. PMID- 11021996 TI - Single amino acid differences are sufficient for CD4(+) T-cell recognition of a heterologous virus by cattle persistently infected with bovine viral diarrhea virus. AB - Cattle that are persistently infected (PI) with one strain of bovine viral diarrhea virus (BVDV) can resolve infection with a second, antigenically heterologous strain but not the homologous strain. Since CD4(+) T cells are thought to be critical for the resolution of acute BVDV infection (Howard et al., 1992, Vet. Immunol. Immunopathol. 32, 303-314), we have examined the recognition of a heterologous virus (NADL) by CD4(+) T cells from Pe515-PI animals. The immune response of non-PI control cattle challenged with NADL or Pe515ncp was strain cross-reactive, whereas Pe515-PI animals responded to NADL only. The immune repertoire of both groups included NS3, which differs by approximately 1% (9/683) amino acids between these two viruses. Lymphoproliferative responses to proteins and synthetic peptides corresponding to three nonconservative differences in NS3 demonstrated that CD4(+) T cells from non-PI control animals responded well to proteins but poorly to the peptides from both viruses. In contrast, PI animals were responsive to heterologous proteins and peptides but nonresponsive to the homologous equivalents. A single amino acid difference between the two sequences was sufficient to allow responsiveness. PMID- 11021997 TI - Modulation of Moloney leukemia virus long terminal repeat transcriptional activity by the murine CD4 silencer in retroviral vectors. AB - We investigated whether CD4 gene regulatory sequences might be useful for developing transcriptionally targeted Moloney murine leukemia virus (Mo-MLV) based retroviral vectors for gene expression specifically in CD4(+) cells. We could modulate Mo-MLV long terminal repeat (LTR) activity by inserting a 438-bp long fragment containing the murine CD4 silencer in the LTR of the vector; both beta-galactosidase and green fluorescent protein reporter gene activities were strongly down-regulated in both murine and human CD8(+) cells, but not in CD4(+) lymphoid cell lines and freshly isolated lymphocytes transduced with this vector, compared with the findings using a control vector carrying wild-type LTRs. Titration experiments on NIH-3T3 cells revealed that inclusion of the CD4 silencer in the LTRs did not reduce the titer of the vectors. These findings indicate that a cellular silencer can be successfully included in retroviral vectors, where it maintains its transcription-regulatory function, thus suggesting a novel approach to transcriptional targeting. PMID- 11021998 TI - Mutations in the E2 glycoprotein of Venezuelan equine encephalitis virus confer heparan sulfate interaction, low morbidity, and rapid clearance from blood of mice. AB - The arbovirus, Venezuelan equine encephalitis virus (VEE), causes disease in humans and equines during periodic outbreaks. A murine model, which closely mimics the encephalitic form of the disease, was used to study mechanisms of attenuation. Molecularly cloned VEE viruses were used: a virulent, epizootic, parental virus and eight site-specific glycoprotein mutants derived from the parental virus. Four of these mutants were selected in vitro for rapid binding and penetration, resulting in positive charge changes in the E2 glycoprotein from glutamic acid or threonine to lysine (N. L. Davis, N. Powell, G. F. Greenwald, L. V. Willis, B. J. Johnson, J. F. Smith, and R. E. Johnston, Virology 183, 20-31, 1991). Tissue culture adaptation also selected for the ability to bind heparan sulfate as evidenced by inhibition of plaque formation by heparin, decreased infectivity for CHO cells deficient for heparan sulfate, and tight binding to heparin-agarose beads. In contrast, the parental virus and three other mutants did not use heparan sulfate as a receptor. All eight mutants were partially or completely attenuated with respect to mortality in adult mice after a subcutaneous inoculation, and the five mutants that interacted with heparan sulfate in vitro had low morbidity (0-50%). These same five mutants were cleared rapidly from the blood after an intravenous inoculation. In contrast, the parental virus and the other three mutants were cleared very slowly. In summary, the five VEE viruses that contain tissue-culture-selected mutations interacted with cell surface heparan sulfate, and this interaction correlated with low morbidity and rapid clearance from the blood. We propose that one mechanism of attenuation is rapid viral clearance in vivo due to binding of the virus to ubiquitous heparan sulfate. PMID- 11021999 TI - Sequence analysis of VP4 and VP7 genes of nontypeable strains identifies a new pair of outer capsid proteins representing novel P and G genotypes in bovine rotaviruses. AB - During a limited epidemiological study, the serotype specificities of several isolates of bovine rotavirus, exhibiting identical electropherotypes, from a single cattle farm near Bangalore, India, could not be determined using a panel of serotyping monoclonal antibodies (MAbs) specific for G serotypes 1-6 and 10. To determine the genotypes of these isolates, the nucleotide sequences of the genes encoding the outer capsid proteins VP4 and VP7 of two representative isolates, Hg18 and Hg23, were determined. The corresponding gene sequences from the two isolates were identical, indicating that these isolates represented a single strain of bovine rotavirus. Comparison of the VP4 nucleotide (nt) and the deduced amino acid (aa) sequences with those of several human and animal rotavirus strains representing all of the currently recognized 20 different VP4 (P) genotypes revealed low nt and aa sequence identities of 61.0 to 74.2% and 57.9 to 78.2% for VP4. The percentages of amino acid homology for the VP8* and VP5* regions of VP4 were 37.7 to 67.9 and 68.1 to 84.2%, respectively. The nt and aa sequences of the VP7 gene were also distinct from those of human and animal strains belonging to the previously established 14 VP7(G) serotypes (65.9 to 75.5% nt and 59.5 to 77.6% aa identities). These findings suggest the classification of the VP4 and VP7 genes of the bovine isolates represented by Hg18 as new P and G genotypes and provide further evidence for the vast genetic/antigenic diversity of group A rotaviruses. PMID- 11022000 TI - Involvement of oxidative stress, NF-IL-6, and RANTES expression in dengue-2-virus infected human liver cells. AB - The liver has been suspected to be one of the major targets of dengue virus infection. Here, we detected increasing secretion of the chemokine RANTES (regulated upon activation, normal T cell expressed and secreted), which functions to recruit the immune cells, in dengue-virus-infected liver cells and patients. Three luciferase reporter genes with various deletions at the 5'-end of the RANTES promoter were constructed to explore the RANTES activation mechanism in human liver cells. The reporter gene was optimally activated by dengue-2 virus when the RANTES promoter contains the region from the transcription starting site (+1) to the nucleotide at the -181 position. NF-IL-6 and an undefined factor forming DNA-protein complexes in the RANTES promoter E and A/B regions in the infected cells were demonstrated by electrophoretic mobility shift assay. Further analysis showed that oxidative stress was an upstream inducer of NF-IL-6 and RANTES signaling in dengue-virus-infected liver cells. This finding was demonstrated by three antioxidants (N-acetyl-l-cysteine, nitro-l-arginine methyl ester, and pyrrolidine dithiocarbamate) used to suppress the activation. In contrast, the DNA binding activity of the undefined factor was not affected by the antioxidant treatment, indicating the existence of an oxidant-independent pathway. We hypothesize that dengue virus infection of the liver cells may trigger both an oxidant-dependent and an oxidant-independent pathway to up regulate RANTES mRNA expression through activating NF-IL-6 and an undefined factor, respectively. In conclusion, the present study suggests a new direction for the study of liver pathogenesis involving RANTES in host immune responses during dengue virus infection. PMID- 11022001 TI - The HCV core protein acts as a positive regulator of fas-mediated apoptosis in a human lymphoblastoid T cell line. AB - Hepatitis C virus (HCV) is a major human pathogen causing mild to severe liver disease worldwide and is remarkably efficient at establishing persistent infections. Previously, we have shown that the core protein has an immunomodulatory function including the suppression of T lymphocyte responses to viral infection. To investigate the underlying mechanism for the role of core protein in immune modulation, we examined the effect of core on the sensitivity of the human T cell line, Jurkat, to Fas-mediated apoptosis. The transient and stable expression of core protein in Jurkat cells increased the sensitivity of cells to Fas-mediated apoptosis when compared to control cells expressing vector DNA alone. In addition, we demonstrated that the core protein binds to the cytoplasmic domain of Fas which may enhance the downstream signaling event of Fas mediated apoptosis. The expression of core protein did not alter the cell surface expression of Fas, indicating that the increased sensitivity of core-expressing cells to Fas ligand was not due to upregulation of Fas. Furthermore, we observed the augmentation of caspase-3 activity in core-expressing cells. These results suggest that the core protein may promote the apoptosis of immune cells during HCV infection via the Fas signaling pathway, thus facilitating HCV persistence. PMID- 11022002 TI - Influenza A and sendai viruses induce differential chemokine gene expression and transcription factor activation in human macrophages. AB - Chemokines regulate leukocyte traffic and extravasation into the site of inflammation. Here we show that influenza A- or Sendai virus-infected human macrophages produce MIP-1alpha, MIP-1beta, RANTES, MCP-1, MCP-3, MIP-3alpha, IP 10, and IL-8, whereas no upregulation of MIP-3beta, eotaxin, or MDC production was detected. Influenza A virus was a better inducer of MCP-1 and MCP-3 production than Sendai virus, whereas MIP-1alpha, MIP-1beta, RANTES, MIP-3alpha, and IL-8 were induced preferentially by Sendai virus. Infection in the presence of protein synthesis inhibitor indicated that ongoing protein synthesis was required for influenza A virus-induced expression of MCP-1, MCP-3, and IP-10 genes, whereas Sendai virus-induced chemokine mRNA expression took place in the absence of de novo protein synthesis. Neutralization of virus-induced IFN alpha/beta resulted in downregulation of virus-induced IP-10, MCP-1, and MCP-3 mRNA expression. IFN-alpha or IFN-gamma were found to directly enhance MCP-1, MCP 3, and IP-10 mRNA expression. Both influenza A and Sendai viruses similarly activated transcription factor NF-kappaB. In contrast to NF-kappaB, IRFs and STATs, the other transcription factors involved in the regulation of chemokine gene expression, were differentially activated by these viruses. Influenza A virus more efficiently activated ISGF3 complex formation and Stat1 DNA-binding compared to Sendai virus, which in turn was a more potent activator of IRF-1. Our results show that during viral infections macrophages predominantly produce monocyte and Th1 cell attracting chemokines. Furthermore, virus-induced IFN alpha/beta enhanced chemokine gene expression in macrophages emphasizing the role of IFN-alpha/beta in the development of Th1 immune responses. PMID- 11022003 TI - Suppressor mutations that allow sindbis virus RNA polymerase to function with nonaromatic amino acids at the N-terminus: evidence for interaction between nsP1 and nsP4 in minus-strand RNA synthesis. AB - The alphavirus RNA polymerase, nsP4, invariably has a Tyr residue at the N terminus. Previously we reported that the N-terminal Tyr residue of nsP4 of Sindbis virus, the type species of the genus Alphavirus, can be substituted with Phe, Trp, or His without altering the wild-type phenotype in cultured cells but that other substitutions tested, except for Met, were lethal or quasilethal. Here we report the identification of two suppressor mutations in nsP4 (Glu-191 to Leu and Glu-315 to Gly, Val, or Lys) and one in nsP1 (Thr-349 to Lys) that allow nsP4 with nonaromatic amino acids at the N-terminus to function at 30 degrees C. The suppressor mutation at nsP4 Glu-315 occurred most frequently. All three suppressor mutations suppressed the effects of Ala, Arg, or Leu at the N-terminus of nsP4 with almost equal efficiency and thus the effect of the suppressing mutation is independent of the nsP4 N-terminal residue. Reconstructed mutants containing nsP1-T349K or nsP4-E315G combined with Ala-nsP4 had a defect in minus strand RNA synthesis at 40 degrees C. A double mutant containing nsP4-Q191L combined with Ala-nsP4 was unstable and could not be tested for RNA synthesis because it reverted to temperature-independence too rapidly. Combinations of nsP1 T349K or nsP4-E315G with Leu, Arg, His, or any aromatic amino acid at the N terminus of nsP4 also made the mutant viruses temperature sensitive. The results from this study and from a previous report on the shutoff of minus-strand RNA synthesis at 40 degrees C with the nsP1-A348T mutation in ts11 suggests that the N-terminus nsP4 interacts with nsP1 during initiation of minus-strand RNA synthesis. PMID- 11022004 TI - Avian hemangioma retrovirus induces cell proliferation via the envelope (env) gene. AB - Several years ago, a field strain retrovirus, avian hemangioma virus (AHV), was isolated from hemangioma tumors in layer hens. Sequence analysis indicated that the AHV genome contains the three prototypic retroviral genes, gag, pol, and env, and is devoid of an oncogene. In cultured endothelial cells, however, AHV induced a significant cytopathic effect through a typical apoptotic cascade. We now demonstrate that AHV also induces cell proliferation and anchorage-independent growth of BSC-1 epithelial cells and NIH-3T3 fibroblasts. This was shown by measurements of (1) cell viability, (2) DNA synthesis, (3) flow cytometry analysis of the cell DNA content, and (4) clonogenic efficiency of the infected cells. Anchorage-independent cell growth was demonstrated by colony formation in soft agar. Moreover, the AHV env gene was cloned into a MuLV-based retroviral vector, and infection of NIH-3T3 cells with this vector induced cell proliferation as well as clonogenic growth. These results suggest that AHV, which is devoid of an oncogene, is a pleiotropic activator capable of inducing either apoptosis or cellular proliferation, depending on the infected cell type. PMID- 11022005 TI - Characterization of human immunodeficiency virus type 1 vif gene in long-term asymptomatic individuals. AB - We have determined the sequence of the human immunodeficiency virus type 1 (HIV 1) vif genes from a cohort of 42 long-term nonprogressors (LTNP) and compared these sequences to those of 8 late progressors. The coding potential of the vif open reading frame directly derived by nested PCR from uncultured peripheral blood mononuclear cell DNA was conserved in all 50 individuals. The nucleotide distances between vif sequences were not significantly different between LTNP and late progressors, indicating similar selections of viruses within both types of long-term HIV-1-infected subjects. However, a statistically significant correlation between an amino acid signature at position 132 of Vif and the viral load was found within LTNP. Namely, amino acid Ser was associated with low viral load and amino acid Arg with high viral load. This signature was also observed when LTNP with low viral load were compared to progressors. The Ser132 signature was introduced in place of Arg132 present in the HIV-1 YU-2 Vif prototype into chimeric viruses to assess the impact of Vif signature on the virus. While the replication properties in the SupT1 cell line were unmodified, the mutagenized virus revealed a fivefold decreased replication in activated PBMC, suggesting a possible role of this Vif signature for viral production in vivo. PMID- 11022006 TI - Satellite and defective RNAs of Cryphonectria hypovirus 3-grand haven 2, a virus species in the family Hypoviridae with a single open reading frame. AB - Cryphonectria parasitica hypovirus 3-Grand Haven 2 (CHV3-GH2) is the most recently characterized member of the Hypoviridae family of viruses associated with hypovirulence of the chestnut blight fungus. Isolates of CHV3-GH2 contain either three or four double-stranded (ds) RNAs that are visible on ethidium bromide-stained agarose or polyacrylamide gels. Only the largest dsRNA appears to be required for virus infectivity, and was characterized previously (C. D. Smart et al., 1999, Virology 265, 66-73). In this study, we report the cloning, sequencing, and analysis of the other three dsRNAs. Sizes of the accessory dsRNAs are 3.6 kb (dsRNA 2), 1.9 kb (dsRNA 3), and 0.9 kb (dsRNA 4), compared to 9.8 kb for the genomic dsRNA segment (dsRNA 1). All three accessory dsRNA species are polyadenylated on the 3'-end of one strand, as is genomic dsRNA. DsRNA 2 represents a defective form of dsRNA 1, with the 5'-terminal 1.4 kb derived from the 5'-end of dsRNA 1 and the 3'-terminal 2.2 kb from the 3'-end of dsRNA 1. A single major open reading frame (ORF) is evident from deduced translations of dsRNA 2. The deduced translation product is a 91-kDa protein that represents a fusion consisting of the entire N-terminal protease and the entire putative helicase domain. DsRNAs 3 and 4 represent satellite RNAs that share very little sequence with dsRNA 1 and 2. DsRNA 4 is 937 nucleotides, excluding the poly(A)(+). The first AUG of the polyadenylated strand of dsRNA 4 occurs eight residues in from the 5'-terminus and would initiate the largest ORF on dsRNA 4, with the coding capacity for a 9.4-kDa protein. Within the deduced ORF and approximately 100 nucleotides from the 5'-end of dsRNA 4 is a 22-base sequence that is identical to sequences found in the nontranslated leaders of dsRNAs 1 and 2. DsRNA 3 accumulation in infected cultures varied, but it was less abundant than dsRNA 4. DsRNA 3 was found to represent a head-to-tail dimer of dsRNA 4 linked by a poly(A)/(U) stretch of 40-70 residues. PMID- 11022007 TI - Comparison of identical temperature-sensitive mutations in the L polymerase proteins of sendai and parainfluenza3 viruses. AB - The L subunit of the RNA-dependent RNA polymerase of negative strand RNA viruses is believed to possess all the enzymatic activities necessary for viral transcription and replication. Mutations in the L proteins of human parainfluenza virus type 3 (PIV3) and vesicular stomatitis virus (VSV) have been shown to confer temperature sensitivity to the viruses; however, their specific defects have not been determined. Mutant PIV3 L proteins expressed from plasmids were tested for temperature sensitivity in transcription and replication in a minigenome reporter system in cells and for in vitro transcription from purified PIV3 template. The single L mutants, Y942H and L992F, were temperature sensitive (ts) in both assays, although viral RNA synthesis was not completely abolished at the nonpermissive temperature. Surprisingly, the T1558I L mutant was not ts, although its cognate virus was ts. Thus the ts defect in this virus may be due to the abrogation of an essential interaction of the mutant polymerase with a host cell component, which is not measured by the RNA synthesis assays. Most of the combinations of the PIV3 L mutations were not additive and did not show temperature sensitivity in in vitro transcription. Since they were ts in the minigenome assay in vivo, replication appears to be specifically defective. The ts mutations in PIV3 and VSV L proteins were also substituted into the Sendai L protein to compare the defects in related systems. Only Sendai Y942H L was ts in both transcription and replication. One Sendai L mutant, L992F, gave much better replication than transcription. Several other mutants could transcribe but not replicate in vitro, while replication in vivo was normal. PMID- 11022008 TI - Evaluation of recombinant vaccinia virus--measles vaccines in infant rhesus macaques with preexisting measles antibody. AB - Immunization of newborn infants with standard measles vaccines is not effective because of the presence of maternal antibody. In this study, newborn rhesus macaques were immunized with recombinant vaccinia viruses expressing measles virus hemagglutinin (H) and fusion (F) proteins, using the replication-competent WR strain of vaccinia virus or the replication-defective MVA strain. The infants were boosted at 2 months and then challenged intranasally with measles virus at 5 months of age. Some of the newborn monkeys received measles immune globulin (MIG) prior to the first immunization, and these infants were compared to additional infants that had maternal measles-neutralizing antibody. In the absence of measles antibody, vaccination with either vector induced neutralizing antibody, cytotoxic T cell (CTL) responses to measles virus and protection from systemic measles infection and skin rash. The infants vaccinated with the MVA vector developed lower measles-neutralizing antibody titers than those vaccinated with the WR vector, and they sustained a transient measles viremia upon challenge. Either maternal antibody or passively transferred MIG blocked the humoral response to vaccination with both WR and MVA, and the frequency of positive CTL responses was reduced. Despite this inhibition of vaccine-induced immunity, there was a reduction in peak viral loads and skin rash after measles virus challenge in many of the infants with preexisting measles antibody. Therefore, vaccination using recombinant vectors such as poxviruses may be able to prevent the severe disease that often accompanies measles in infants. PMID- 11022009 TI - Functional features of hepatitis C virus glycoproteins for pseudotype virus entry into mammalian cells. AB - We have previously reported the generation of pseudotype virus from chimeric gene constructs encoding the ectodomain of the E1 or E2 glycoprotein of hepatitis C virus (HCV) genotype 1a appended to the trans membrane domain and cytoplasmic tail of the vesicular stomatitis virus (VSV) G protein. Sera derived from chimpanzees immunized with homologous HCV glycoproteins neutralized pseudotype virus infectivity (L. M. Lagging et al., J. Virol. 72, 3539-3546, 1998). We have now extended this study to further understand the role of HCV glycoproteins in pseudotype virus entry. Although a number of mammalian epithelial cells were susceptible to VSV/HCV pseudotype virus infection, plaquing efficiency was different among host cell lines. Pseudotype virus adsorption at low temperature decreased plaque numbers. Treatment of E1 or E2 pseudotype virus in media between pH 5 and 8 before adsorption on cells did not significantly reduce plaque numbers. On the other hand, treatment of cells with lysosomotropic agents or inhibitors of vacuolar H(+) ATPases had an inhibitory role on virus entry. Concanavalin A, a plant lectin, exhibited neutralization of both HCV E1 and E2 pseudotype virus infectivity. However, mannose binding protein, a C-type mammalian lectin, did not neutralize virus in the absence or presence of serum complement. Pseudotype virus infectivity was only partially inhibited by heparin, a highly sulfated glycosaminoglycan, in a saturable manner. Additional studies suggested that low-density lipoprotein receptor related molecules partially inhibit E1 pseudotype virus infectivity, while CD81 related molecules interfere with E2 pseudotype virus infectivity. A further understanding of HCV entry and strategies appropriate for mimicking cell surface molecules may help in the development of new therapeutic modalities against HCV infection. PMID- 11022010 TI - A major locus for myoclonus-dystonia maps to chromosome 7q in eight families. AB - Myoclonus-dystonia (M-D) is an autosomal dominant disorder characterized by myoclonic and dystonic muscle contractions that are often responsive to alcohol. The dopamine D2 receptor gene (DRD2) on chromosome 11q has been implicated in one family with this syndrome, and linkage to a 28-cM region on 7q has been reported in another. We performed genetic studies, using eight additional families with M D, to assess these two loci. No evidence for linkage was found for 11q markers. However, all eight of these families showed linkage to chromosome 7 markers, with a combined multipoint LOD score of 11.71. Recombination events in the families define the disease gene within a 14-cM interval flanked by D7S2212 and D7S821. These data provide evidence for a major locus for M-D on chromosome 7q21. PMID- 11022011 TI - A second-generation genomewide screen for asthma-susceptibility alleles in a founder population. AB - A genomewide screen for asthma- and atopy-susceptibility loci was conducted, using 563 markers, in 693 Hutterites who are members of a single 15-generation pedigree, nearly doubling the sample size from the authors' earlier studies. The resulting increase in power led to the identification of 23 loci in 18 chromosomal regions showing evidence for linkage that is, in general, 10-fold more significant (P<.001 vs. P<.01) than the linkages reported previously in this population. Moreover, linkages to loci in 11 chromosomal regions were identified for the first time in the Hutterites in this report, including five regions (5p, 5q, 8p, 14q, and 16q) showing evidence both of linkage, by the likelihood ratio (LR) chi(2), and of disequilibrium, by the transmission/disequilibrium test. A region on chromosome 19 continues to show evidence for linkage, by both tests, in this study. Studies of 17 candidate genes provide evidence for association with variation in the IL4RA gene (16p12), the HLA class II genes (6p21), and the interferon-alpha gene cluster (9p22), but the lack of evidence for linkage in these regions by the LR chi(2) test suggests that these are minor susceptibility loci. A polymorphism in the CD14 gene is in linkage disequilibrium with an as yet unidentified susceptibility allele in the 5q cytokine cluster, a region showing evidence for linkage among the Hutterites. Finally, 10 of the regions showing evidence for linkage in the Hutterites have shown evidence of linkage to related phenotypes in other genome screens, suggesting that these regions may contain common alleles that have relatively large effects on asthma and atopy phenotypes in diverse populations. PMID- 11022012 TI - Autosomal recessive cerebellar ataxia with oculomotor apraxia (ataxia telangiectasia-like syndrome) is linked to chromosome 9q34. AB - Ataxia with oculomotor apraxia (ataxia-telangiectasia-like syndrome [AOA]; MIM 208920) is an autosomal recessive disorder characterized by ataxia, oculomotor apraxia, and choreoathetosis. These neurological features resemble those of ataxia-telangiectasia (AT), but in AOA there are none of the extraneurological features of AT, such as immunodeficiency, neoplasia, chromosomal instability, or sensitivity to ionizing radiation. It is unclear whether these patients have a true disorder of chromosomal instability or a primary neurodegenerative syndrome, and it has not been possible to identify the defective gene in AOA, since the families have been too small for linkage analysis. We have identified a new family with AOA, and we show that the patients have no evidence of chromosomal instability or sensitivity to ionizing radiation, suggesting that AOA in this family is a true primary cerebellar ataxia. We have localized the disease gene, by linkage analysis and homozygosity mapping, to a 15.9-cM interval on chromosome 9q34. This work will ultimately allow the disease gene to be identified and its relevance to other types of autosomal recessive cerebellar ataxias to be determined. PMID- 11022013 TI - The effects of moderate alcohol consumption on female hormone levels and reproductive function. AB - Studies that have investigated the effect of moderate alcohol consumption on the level of oestrogens and progesterone in both pre- and post-menopausal women are reviewed. It is concluded that several lines of evidence point to an alcohol induced rise in natural or synthetic oestrogen levels in women. Proposed mechanisms include an increased rate of aromatization of testosterone or a decreased rate of oxidation of oestradiol to oestrone. Moderate alcohol consumption has also been linked to decreased progesterone levels in pre menopausal women. The relevance of these findings to female health, fertility and the timing of the menopause is considered. PMID- 11022014 TI - Effect of methylene blue on the disposition of ethanol. AB - The effect of methylene blue on the disposition of ethanol was studied in rats and humans. Methylene blue increased the metabolism of [(14)C]ethanol to (14)CO(2) in isolated hepatocytes and in intact rats by 75% and 30%, respectively. In healthy volunteers, methylene blue did not affect the pharmacokinetics of ethanol and did not alleviate the ethanol-induced NAD redox changes as reflected by the increase in the [lactate]/[pyruvate] ratio. PMID- 11022015 TI - Rat liver tryptophan pyrrolase activity and gene expression during alcohol withdrawal. AB - Rat liver tryptophan (Trp) pyrrolase activity and gene expression were studied in relation to the alcohol-withdrawal syndrome (AWS). Both activity and gene expression were enhanced after withdrawal of ethanol-containing liquid diets and the time-course of these changes mirrored that of development and intensity of the behavioural disturbances of the AWS. By contrast, no correlation was observed between the AWS-induced behaviour and changes in activity of another hepatic glucocorticoid-inducible enzyme, tyrosine aminotransferase, and a negative correlation was noted between behaviour and the gene expression of this latter enzyme and also of that of the hepatic glucocorticoid receptor. We suggest that the metabolic consequences of activation of liver Trp pyrrolase during alcohol withdrawal may play a role in the behavioural features of the AWS. PMID- 11022016 TI - Paradigm to test a drug-induced aversion to ethanol. AB - The screening of new agents for aversive therapy of alcoholism requires a simple animal model. Animals trained to ingest ethanol solutions and subsequently administered a drug known to produce an aversion to ethanol in humans, do not readily make the association between the malaise induced by the aversive drug ethanol reaction and the consumption of the same ethanol-containing solution that has been consumed previously without ill effects. An experimental paradigm is reported in which the malaise of the drug-ethanol reaction is quickly recognized by rats as derived from ethanol. Disulfiram was used as the model drug. Lewis rats were deprived of water for 18 h after which 6% (v/v) ethanol was offered as the only fluid. During the first hour of ethanol access, both controls (vehicle) and disulfiram (100 mg/kg)-treated animals consumed intoxicating amounts of ethanol (0.7-0.9 g ethanol/kg). Plasma acetaldehyde levels developed were 3-5 microM and 40-50 microM in the two groups respectively. After this time, disulfiram-treated animals virtually ceased consuming alcohol (90% inhibition), indicating that the disulfiram-ethanol reaction is associated with alcohol ingestion. Control animals continued consuming the alcohol solution for the additional 4-5 h tested. This model should be of value in the testing of new agents that reduce aldehyde dehydrogenase levels for prolonged periods for their potential as an aversive treatment in alcoholism. PMID- 11022017 TI - Effect of short-term ethanol administration on cadmium retention and bioelement metabolism in rats continuously exposed to cadmium. AB - The present study was performed to assess the effect of short-term ethanol administration on cadmium retention and accumulation as well as on bioelement metabolism (zinc, copper, calcium, and magnesium) in rats exposed to an aqueous solution of cadmium chloride for 8 weeks. Intoxication with cadmium led to accumulation of this toxic metal, particularly in the liver and kidney, which was linked to metallothionein synthesis as well as to a disturbance in the metabolism of zinc, copper, and calcium. These effects were dependent on the level of exposure. The administration of ethanol in the final phase of cadmium treatment increased cadmium retention and accumulation in the body with simultaneous elevation in liver and kidney metallothionein concentration. Ethanol alone or with cadmium caused or intensified the cadmium-induced changes in metabolism of zinc and copper. Calcium metabolism disturbed by cadmium was not influenced by ethanol. Neither agents had any effect on magnesium metabolism. We conclude that even short-term ethanol consumption in conditions of exposure to cadmium can increase this heavy metal body burden and lead to more serious disturbances in metabolism of important elements such as zinc and copper. Cadmium- and ethanol induced changes in the homeostasis of these microelements are probably connected with the ability of both xenobiotics to cause metallothionein induction. PMID- 11022018 TI - Forced ethanol treatment stimulates and inhibits ethanol intake in a rat model of alcoholism. AB - In a model of psychological dependence, a very stable ethanol intake was induced by a chronic (1-year) intermittent (once a week) exposure to intoxicating amounts of ethanol (24 h choice between ethanol and water, followed by 2.0 g/kg i.p.). After this year, the rats had continuous access to ethanol and water. Stability was shown by the ability of the rats to take the same dose of ethanol (in g/kg) when the concentration was changed from 10 to 20%. To study possible priming or inhibiting effects on ethanol intake, ethanol was injected i.p., first as 20%, 40% or 60% of the intake in the 24 h prior to the injection, then as fixed doses of 0.5, 1, and 2 g/kg, and the ethanol intake during the following 24-h period was recorded. The results showed that, following a low dose of ethanol, voluntary ethanol intake was increased in rats with a low, and decreased in rats with a high, ethanol intake, while high doses of ethanol seemed to decrease voluntary ethanol intake in all rats. The results are discussed in relation to theories about loss of control of drinking and relapse in humans. PMID- 11022019 TI - Low levels of ethanol stimulate and high levels decrease phosphorylation in microtubule-associated proteins in rat brain: an in vitro study. AB - Phosphorylation and dephosphorylation of proteins associated with microtubules (MAPs) modulate the functional properties of microtubules (MT). A study was designed to test the hypothesis that ethanol at pharmacologically relevant levels affects phosphorylation of MAPs. Low (6, 12, 24, and 48 mM) and high (96, 384, and 768 mM) levels of ethanol were used in the study. MT prepared from rat brain by successive cycles of assembly-disassembly were found to contain two high molecular weight proteins (MAP2 and MAP1), tubulin, and 70-kDa neurofilament. The kinase activity was determined using [gamma(32)P]ATP as a phosphate donor. The results showed that ethanol primarily stimulated MAP2 phosphorylation. Low levels of ethanol stimulated, whereas high levels decreased, the kinase activity. MAP1 was phosphorylated to a lesser extent. 70-kDa neurofilament and tubulin were phosphorylated, however, the dose-dependent biphasic effect of ethanol on phosphorylation was not found in these cytoskeleton proteins. To determine whether the ethanol-induced kinase activity was cAMP-dependent, the catalytic subunit of cAMP-dependent protein kinase was isolated, purified, and kinase activity was determined with and without ethanol. The results showed that cAMP was not involved in ethanol-induced kinase activity. We conclude that ethanol predominantly stimulates phosphorylation of MAP2 in a dose-dependent manner. PMID- 11022020 TI - Evidence of acetaldehyde-protein adduct formation in rat brain after lifelong consumption of ethanol. AB - Acetaldehyde, the first metabolite of ethanol, has been shown to be capable of binding covalently to liver proteins in vivo, which may be responsible for a variety of toxic effects of ethanol. Acetaldehyde-protein adducts have previously been detected in the liver of patients and experimental animals with alcoholic liver disease. Although a role for acetaldehyde as a possible mediator of ethanol induced neurotoxicity has also been previously suggested, the formation of protein-acetaldehyde adducts in brain has not been examined. This study was designed to examine the occurrence of acetaldehyde-protein adducts in rat brain after lifelong ethanol exposure. A total of 27 male rats from the alcohol preferring (AA) and alcohol-avoiding (ANA) lines were used. Four ANA rats and five AA rats were fed 10-12% (v/v) ethanol for 21 months. Both young (n = 10) and old (n = 8) rats receiving water were used as controls. Samples from frontal cortex, cerebellum and liver were processed for immunohistochemical detection of acetaldehyde adducts. In four (two ANA, two AA rats) of the nine ethanol-exposed rats, weak or moderate positive reactions for acetaldehyde adducts could be detected both in the frontal cortex and cerebellum, whereas no such immunostaining was found in the remaining five ethanol-treated rats or in the control rats. The positive reaction was localized to the white matter and some large neurons in layers 4 and 5 of the frontal cortex, and to the molecular layer of the cerebellum. Interestingly, the strongest positive reactions were found among the ANA rats, which are known to display high acetaldehyde levels during ethanol oxidation. We suggest that acetaldehyde may be involved in ethanol induced neurotoxicity in vivo through formation of adducts with brain proteins and macromolecules. PMID- 11022021 TI - Attrition in a follow-up study of driving while impaired offenders: who is lost? AB - High attrition rates seriously threaten the validity of follow-up studies of criminal justice populations. This study examines attrition from a follow-up study of drink-driving offenders referred 5 years earlier to a screening programme. The aim of the study was to determine which factors are most closely associated with: (1) inability to locate subjects, (2) subjects' refusal to participate; (3) the manner in which subjects refuse. Logistic regression models compared the following groups of subjects: located vs not located; interviewed vs not interviewed; type of refusal (direct vs indirect). Independent variables included gender, age group, ethnicity, whether the subject had a telephone, compliance with and completion of the screening programme, alcohol dependence or abuse diagnosis vs no diagnosis, breath-alcohol level (BAL) at the time of arrest, and whether the subject had an outstanding arrest warrant. Some factors (younger age, screening compliance, Mexican national ethnicity, and having an outstanding arrest warrant) predicted both inability to locate and type of refusal. Hispanic ethnicity and having a telephone predicted better success with locating subjects. Among refusers, non-Hispanic whites were more likely than other ethnic groups to refuse directly, and those with warrants were more likely to refuse indirectly. Non-compliance with the screening programme was also associated with differential follow-up rates. Neither arrest BAL nor alcohol diagnoses was associated with differential rates of follow-up. We conclude that alcohol diagnosis does not appear to influence successful follow-up in this criminal justice population. Rather, tracking and interviewing challenges differed among ethnic groups, suggesting a need for culturally sensitive recruitment strategies in these populations. PMID- 11022022 TI - The effects of alcohol intoxication on aggressive responses in men and women. AB - A considerable literature, clinical and experimental, has demonstrated the aggression-eliciting effects of alcohol intoxication. However, the focus of the experimental literature has been primarily on men and the studies on women have been inconclusive. This study was conducted to test for possible gender differences in the manifestation of alcohol-induced aggression. Participants were 54 males and 60 females, aged 18-30 years, who competed in a competitive aggression paradigm either sober or intoxicated. As expected, intoxicated men were more aggressive than their sober peers. However, under high provocation, both sober and intoxicated, women manifested aggression comparable to the intoxicated men. This study suggests that women can be as aggressive as men, and that alcohol intoxication does not seem to be as important a determining factor. PMID- 11022023 TI - The long-term cost-effectiveness of improving alcohol abstinence with adjuvant acamprosate. AB - A computer model was developed with decision analysis software to explore the long-term clinical and economic outcomes of alcohol abstinence maintenance with either standard counselling therapy or standard therapy plus 48 weeks of adjuvant acamprosate in detoxified alcoholic patients. Important complications of alcoholism were modelled using Markov processes, and included relapse (return to drinking), alcohol-related hepatic disease, acute and chronic pancreatitis, acute and chronic gastritis, oropharyngeal carcinoma, oesophageal carcinoma, alcoholic cardiomyopathy, alcohol-related peripheral neuropathy, alcoholic psychosis, accidental death, and suicide. Probabilities of developing complications were dependent on whether the patients within the cohort remained abstinent or had relapsed. Relapse rates, probabilities, and costs for acamprosate therapy and treatment of complications were taken from published literature. The analysis was performed from the German health insurance perspective. Life expectancy and total lifetime costs (costs of initial abstinence maintenance therapy plus costs of complications) were calculated for a typical male cohort with average age of 41 years, 80% with fatty liver, 15% with cirrhosis, 22% with chronic pancreatitis, and 1% with alcoholic cardiomyopathy at baseline. Life expectancy with and without acamprosate therapy was 15.90 and 14.70 years respectively, and discounted (5% per annum) average total lifetime costs per patient were DEM 46 448 and DEM 49 549 respectively. We conclude that, despite the acquisition costs of DEM 2177, adjuvant acamprosate therapy was both clinically and economically attractive under conservative assumptions. PMID- 11022024 TI - Alcoholics with the dopamine receptor DRD2 A1 allele have lower platelet monoamine oxidase-B activity than those with the A2 allele: a preliminary study. AB - Low platelet monoamine oxidase B (MAO-B) activity and the presence of the Taq1 A1 allele of the dopamine D2 receptor (DRD2) gene have independently been proposed as 'biological/genetic' markers for alcoholism. In the present study, the relationship between these two markers was investigated in a group of socially stable Caucasian middle-aged men with a mean (+/-SD) daily ethanol consumption of 85 +/- 57 g. The platelet MAO-B activity was significantly lower in individuals with the DRD2 A1 allele (n = 8), compared to those without it (n = 29). This relationship remained unchanged when including only subjects who fulfilled DSM-IV criteria for alcohol dependence (n = 27). The finding suggests that alcoholics who are carriers of the DRD2 A1 allele may have lower platelet MAO-B activity. PMID- 11022025 TI - Fetal alcohol syndrome (FAS) primary prevention through FAS diagnosis: I. Identification of high-risk birth mothers through the diagnosis of their children. AB - A 5-year, fetal alcohol syndrome (FAS) primary prevention study was conducted in Washington State to: (1) assess the feasibility of using a FAS diagnostic and prevention clinic as a centre for identifying and targeting primary prevention intervention to high-risk women (namely women who had given birth to a child with FAS); (2) generate a comprehensive, lifetime profile of these women; (3) identify factors that have enhanced and/or hindered their ability to achieve abstinence. The results of this study are presented in two parts: work on objective 1 is summarized in the present paper; whereas that on objectives 2 and 3 is summarized in the accompanying paper. This project demonstrated that a multidisciplinary FAS Diagnostic and Prevention Network (FAS DPN) clinic could successfully attract and meet the diagnostic and treatment planning needs of patients presenting with prenatal alcohol exposure. One out of every three patients evaluated in the FAS DPN clinics was diagnosed with FAS or static encephalopathy/alcohol exposed. The birth mothers of one out of every three of these children diagnosed with FAS or static encephalopathy/alcohol exposed could be located and directly contacted. Half of the birth mothers directly contacted were still at risk for producing more children damaged by prenatal alcohol exposure. Thus, one out of every 18 children evaluated in the FAS DPN clinics had a birth mother who could be found and was at risk of producing more children damaged by prenatal alcohol exposure. Primary prevention programmes targeted to this high-risk population could lead to measurable, cost-effective reductions in the incidence of FAS. Using this approach, the cost of raising a child with FAS would be roughly 30 times the cost of preventing FAS in the child. The benefit to the children, their mothers, and society would be immeasurable. PMID- 11022026 TI - Fetal alcohol syndrome (FAS) primary prevention through fas diagnosis: II. A comprehensive profile of 80 birth mothers of children with FAS. AB - A 5-year, fetal alcohol syndrome (FAS) primary prevention study was conducted in Washington State to: (1) assess the feasibility of using a FAS diagnostic and prevention clinic as a centre for identifying and targeting primary prevention intervention to high-risk women; (2) generate a comprehensive, lifetime profile of these women; (3) identify factors that have enhanced and/or hindered their ability to achieve abstinence. The results of this study are presented in two parts. Objective 1 is summarized in the preceding paper and objectives 2 and 3 are summarized here. Comprehensive interviews were conducted with 80 women, who had given birth to a child diagnosed with FAS, to document their sociodemographics, reproductive and family planning history, social and healthcare utilization patterns, adverse social experiences, social support network, alcohol use and treatment history, mental health, and intelligence quotient (IQ). These high-risk women were diverse in racial, educational and economic backgrounds, were often victims of abuse, and challenged by mental health issues. Despite their rather harsh psychosocial profile, many demonstrated the ability to overcome their alcohol dependence over time. Relative to the women who had not achieved abstinence, the women who had achieved abstinence had significantly higher IQs, higher household incomes, larger more satisfactory social support networks, were more likely to report a religious affiliation, and were more likely to be receiving mental health treatment for their mental health disorders. The rate of unintended pregnancies and alcohol-exposed pregnancies was substantial. Key barriers to achieving effective family planning were maternal alcohol and drug use, lack of access to birth control and lack of support by their partner to use birth control. A FAS diagnostic and prevention clinic can be used to identify women at high risk for producing children damaged by prenatal alcohol exposure. Primary prevention programmes targeted to this population could lead to measurable reductions in the incidence of FAS. PMID- 11022027 TI - Drinking patterns among Swedish women: results from a 5-year follow-up of a population-based study. AB - The aims of this study were to: (1) document women's alcohol use over a 5-year period; (2) compare different measures of alcohol consumption such as high alcohol consumption (HAC) and high episodic drinking (HED); (3) to follow the incidence and course of alcohol dependence and abuse (ADA). The study is part of a longitudinal general population-based study, Women and Alcohol in Goteborg. From a stratified random sample of 479 respondents, 399 were interviewed in the first wave (1989/90). In the second wave (1995/96), 386 of the 479 women were available for a follow-up interview. The 12-month prevalence of ADA was 1.3% in both waves. In the longitudinal analysis the 5-year cumulative incidence of ADA was 0.1%. Two new cases of ADA were found. The prevalence of HAC and HED decreased in women of all ages except HED in women born in 1955. We conclude that the 12-month prevalence of ADA remained the same at the 5-year follow-up and that, in contrast to what has been found in some other studies, there was an overall decrease in HAC and in HED. PMID- 11022028 TI - Implementation and dissemination of methods for prevention of alcohol problems in primary health care: a feasibility study. AB - Secondary prevention of alcohol problems in health care has been proved efficacious in many studies, yet its implementation remains scarce, and its effectiveness in regular health care remains unknown. This article reports results from a feasibility study of dissemination of alcohol prevention methods in primary health care in Stockholm. Initial interviews with general practitioners (GPs) and district health nurses indicated that few raised the issue of alcohol with patients, made notes about alcohol in patient charts or found working with alcohol issues rewarding. The impact of a training session, where a project nurse visited all willing GPs and nurses, was limited. Although the uptake of the prevention package was high, follow-up at 3 months indicated that little use was made of the materials. Specifically, screening rates were low. In the future, secondary prevention of alcohol problems will require better adaptation to the realities of primary care. PMID- 11022029 TI - The volume of the liver in patients correlates to body weight and alcohol consumption. PMID- 11022030 TI - Identification of UvrY as the cognate response regulator for the BarA sensor kinase in Escherichia coli. AB - BarA is a membrane-associated protein that belongs to a subclass of tripartite sensors of the two-component signal transduction system family. In this study, we report that UvrY is the cognate response regulator for BarA of Escherichia coli. This conclusion is based upon homologies with analogous two-component systems and demonstrated by both biochemical and genetic means. We show that the purified BarA protein is able to autophosphorylate when incubated with [gamma-(32)P]ATP but not with [alpha-(32)P]ATP or [gamma-(32)P]GTP. Phosphorylated BarA, in turn, acts as an efficient phosphoryl group donor to UvrY but not to the non-cognate response regulators ArcA, PhoB, or CpxR. The specificity of the transphosphorylation reaction is further supported by the fact that UvrY can receive the phosphoryl group from BarA-P but not from the non-cognate tripartite sensor ArcB-P or ATP. In addition, genetic evidence that BarA and UvrY mediate the same signal transduction pathway is provided by the finding that both uvrY and barA mutant strains exhibit the same hydrogen peroxide hypersensitive phenotype. These results provide the first biochemical evidence as well as genetic support for a link between BarA and UvrY, suggesting that the two proteins constitute a new two-component system for gene regulation in Escherichia coli. PMID- 11022031 TI - Identification of a membrane-associated cysteine protease with possible dual roles in the endoplasmic reticulum and protein storage vacuole. AB - SH-EP is a vacuolar cysteine proteinase from germinated seeds of Vigna mungo. The enzyme has a C-terminal propeptide of 1 kDa that contains an endoplasmic reticulum (ER) retention signal, KDEL. The KDEL-tail has been suggested to function to store SH-EP as a transient zymogen in the lumen of the ER, and the C terminal propeptide was thought to be removed within the ER or immediately after exit from the ER. In the present study, a protease that may be involved in the post-translational processing of the C-terminal propeptide of SH-EP was isolated from the microsomes of cotyledons of V. muno seedlings. cDNA sequence for the protease indicated that the enzyme is a member of the papain superfamily. Immunocytochemistry and subcellular fractionation of cotyledon cells suggested that the protease was localized in both the ER and protein storage vacuoles as enzymatically active mature form. In addition, protein fractionations of the cotyledonary microsome and Sf9 cells expressing the recombinant protease indicated that the enzyme associates with the microsomal membrane on the luminal side. The protease was named membrane-associated cysteine protease, MCP. The possibility that a papain-type enzyme, MCP, exists as mature enzyme in both ER and protein storage vacuoles will be discussed. PMID- 11022032 TI - Redox changes of cytochrome b(559) in the presence of plastoquinones. AB - We have found that short chain plastoquinones effectively stimulated photoreduction of the low potential form of cytochrome b(559) and were also active in dark oxidation of this cytochrome under anaerobic conditions in Triton X-100-solubilized photosystem II (PSII) particles. It is also shown that molecular oxygen competes considerably with the prenylquinones in cytochrome b(559) oxidation under aerobic conditions, indicating that both molecular oxygen and plastoquinones could be electron acceptors from cytochrome b(559) in PSII preparations. alpha-Tocopherol quinone was not active in the stimulation of cytochrome photoreduction but efficiently oxidized it in the dark. Both the observed photoreduction and dark oxidation of the cytochrome were not sensitive to 3-(3,4-dichlorophenyl)-1, 1-dimethylurea. It was concluded that both quinone binding sites responsible for the redox changes of cytochrome b(559) are different from either the Q(A) or Q(B) site in PSII and represent new quinone binding sites in PSII. PMID- 11022033 TI - Localization of sequences within the C-terminal domain of the cystic fibrosis transmembrane conductance regulator which impact maturation and stability. AB - Some disease-associated truncations within the 100-residue domain C-terminal of the second nucleotide-binding domain destabilize the mature protein (Haardt, M., Benharouga, M., Lechardeur, D., Kartner, N., and Lukacs, G. L. (1999) J. Biol. Chem. 274, 21873-21877). We now have identified three short oligopeptide regions in the C-terminal domain which impact cystic fibrosis transmembrane conductance regulator (CFTR) maturation and stability in different ways. A highly conserved hydrophobic patch (region I) formed by residues 1413-1416 (FLVI) was found to be crucial for the stability of the mature protein. Nascent chain stability was severely decreased by shortening the protein by 81 amino acids (1400X). This accelerated degradation was sensitive to proteasome inhibitors but not influenced by brefeldin A, indicating that it occurred at the endoplasmic reticulum. The five residues at positions 1400 to 1404 (region II) normally maintain nascent CFTR stability in a positional rather than a sequence-specific manner. A third modulating region (III) constituted by residues 1390 to 1394 destabilizes the protein. Hence the nascent form regains stability on further truncation back to residues 1390 or 1380, permitting some degree of maturation and a low level of cyclic AMP-stimulated chloride channel activity at the cell surface. Thus while not absolutely essential, the C-terminal domain strongly modulates the biogenesis and maturation of CFTR. PMID- 11022034 TI - L-ascorbic acid potentiates endothelial nitric oxide synthesis via a chemical stabilization of tetrahydrobiopterin. AB - Ascorbic acid has been shown to stimulate endothelial nitric oxide (NO) synthesis in a time- and concentration-dependent fashion without affecting NO synthase (NOS) expression or l-arginine uptake. The present study investigates if the underlying mechanism is related to the NOS cofactor tetrahydrobiopterin. Pretreatment of human umbilical vein endothelial cells with ascorbate (1 microm to 1 mm, 24 h) led to an up to 3-fold increase of intracellular tetrahydrobiopterin levels that was concentration-dependent and saturable at 100 microm. Accordingly, the effect of ascorbic acid on Ca(2+)-dependent formation of citrulline (co-product of NO) and cGMP (product of the NO-activated soluble guanylate cyclase) was abolished when intracellular tetrahydrobiopterin levels were increased by coincubation of endothelial cells with sepiapterin (0.001-100 microm, 24 h). In contrast, ascorbic acid did not modify the pterin affinity of endothelial NOS, which was measured in assays with purified tetrahydrobiopterin free enzyme. The ascorbate-induced increase of endothelial tetrahydrobiopterin was not due to an enhanced synthesis of the compound. Neither the mRNA expression of the rate-limiting enzyme in tetrahydrobiopterin biosynthesis, GTP cyclohydrolase I, nor the activities of either GTP cyclohydrolase I or 6-pyruvoyl tetrahydropterin synthase, the second enzyme in the de novo synthesis pathway, were altered by ascorbate. Our data demonstrate that ascorbic acid leads to a chemical stabilization of tetrahydrobiopterin. This was evident as an increase in the half-life of tetrahydrobiopterin in aqueous solution. Furthermore, the increase of tetrahydrobiopterin levels in intact endothelial cells coincubated with cytokines and ascorbate was associated with a decrease of more oxidized biopterin derivatives (7,8-dihydrobiopterin and biopterin) in cells and cell supernatants. The present study suggests that saturated ascorbic acid levels in endothelial cells are necessary to protect tetrahydrobiopterin from oxidation and to provide optimal conditions for cellular NO synthesis. PMID- 11022035 TI - Characterization of a new member of the fatty acid-binding protein family that binds all-trans-retinol. AB - Cellular retinol-binding protein, type I (CRBP-I) and type II (CRBP-II) are the only members of the fatty acid-binding protein (FABP) family that process intracellular retinol. Heart and skeletal muscle take up postprandial retinol but express little or no CRBP-I or CRBP-II. We have identified an intracellular retinol-binding protein in these tissues. The 134-amino acid protein is encoded by a cDNA that is expressed primarily in heart, muscle and adipose tissue. It shares 57 and 56% sequence identity with CRBP-I and CRBP-II, respectively, but less than 40% with other members of the FABP family. In situ hybridization demonstrates that the protein is expressed at least as early as day 10 in developing heart and muscle tissue of the embryonic mouse. Fluorescence titrations of purified recombinant protein with retinol isomers indicates binding to all-trans-, 13-cis-, and 9-cis-retinol, with respective K(d) values of 109, 83, and 130 nm. Retinoic acids (all-trans-, 13-cis-, and 9-cis-), retinals (all trans-, 13-cis-, and 9-cis-), fatty acids (laurate, myristate, palmitate, oleate, linoleate, arachidonate, and docosahexanoate), or fatty alcohols (palmityl, petrosenlinyl, and ricinolenyl) fail to bind. The distinct tissue expression pattern and binding specificity suggest that we have identified a novel FABP family member, cellular retinol-binding protein, type III. PMID- 11022036 TI - Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. AB - GADD45, MyD118, and CR6 (also termed GADD45alpha, beta, and gamma) comprise a family of genes that encode for related proteins playing important roles in negative growth control, including growth suppression. Data accumulated suggest that MyD118/GADD45/CR6 serve similar but not identical functions along different apoptotic and growth suppressive pathways. It is also apparent that individual members of the MyD118/GADD45/CR6 family are differentially induced by a variety of genetic and environmental stress agents. The MyD118, CR6, and GADD45 proteins were shown to predominantly localize within the cell nucleus. Recently, we have shown that both MyD118 and GADD45 interact with proliferating cell nuclear antigen (PCNA), a protein that plays a central role in DNA replication, DNA repair, and cell cycle progression, as well as with the universal cyclin dependent kinase inhibitor p21. In this work we show that also CR6 interacts with PCNA and p21. Moreover, it is shown that CR6 interacts with PCNA via a domain that also mediates interaction of both GADD45 and MyD118 with PCNA. Importantly, evidence has been obtained that interaction of CR6 with PCNA impedes the function of this protein in negative growth control, similar to observations reported for MyD118 and GADD45. PMID- 11022037 TI - Functional interaction of NF-Y and Sp1 is required for type a natriuretic peptide receptor gene transcription. AB - The vasorelaxant and anti-mitogenic activities of the atrial and brain natriuretic peptides depend upon their binding to the type A natriuretic peptide receptor (NPR-A) expressed on the surface of vascular cells. Intervention strategies aimed at controlling NPR-A expression are limited by the paucity of studies in this area. Here we identify a sequence CCAAT between -141 and -137 of the NPR-A promoter that, when mutated, reduces promoter activity by 90% in rat aortic smooth muscle (RASM) cells. Protein/DNA cross-linking and immunoperturbation of electrophoretically shifted complexes formed between RASM nuclear extracts and an oligonucleotide surrounding the CCAAT sequence indicates that the heterotrimeric transcription factor NF-Y binds specifically to the wild type, but not mutated, CCAAT element. Cotransfection of a dominant negative mutant of the NF-YA subunit results in a concentration-dependent decrease in the activity of the NPR-A promoter in RASM cells confirming that endogenous NF-Y is an activator of the promoter. Mutation of the CCAAT element, in conjunction with mutation of all three Sp1 sites previously shown to be involved in NPR-A promoter regulation, virtually eliminates NPR-A promoter activity in RASM cells. Coexpression of all three NF-Y subunits together with Sp1 in Drosophila cells deficient in these factors indicates that NF-Y and Sp1 act synergistically to reconstitute NPR-A promoter activity. A direct physical association between NF-Y and Sp1 can be demonstrated both in vitro by glutathione S-transferase pull-down assay and in the intact cell by coimmunoprecipitation and functional studies. Together, these studies show that NPR-A promoter activity is dominantly regulated through functional, and possibly physical, interactions of NF-Y and Sp1. PMID- 11022038 TI - A decrease in remodeling accounts for the accumulation of arachidonic acid in murine mast cells undergoing apoptosis. AB - The goal of this study was to examine arachidonic acid (AA) metabolism by murine bone marrow-derived mast cells (BMMC) during apoptosis induced by cytokine depletion. BMMC deprived of cytokines for 12-48 h displayed apoptotic characteristics. During apoptosis, levels of AA, but not other unsaturated fatty acids, correlated with the percentage of apoptotic cells. A decrease in both cytosolic phospholipase A(2) expression and activity indicated that cytosolic phospholipase A(2) did not account for AA mobilization during apoptosis. Free AA accumulation is also unlikely to be due to decreases in 5-lipoxygenase and/or cyclooxygenase activities, since BMMC undergoing apoptosis produced similar amounts of leukotriene B(4) and significantly greater amounts of PGD(2) than control cells. Arachidonoyl-CoA synthetase and CoA-dependent transferase activities responsible for incorporating AA into phospholipids were not altered during apoptosis. However, there was an increase in arachidonate in phosphatidylcholine (PC) and neutral lipids concomitant with a 40.7 +/- 8.1% decrease in arachidonate content in phosphatidylethanolamine (PE), suggesting a diminished capacity of mast cells to remodel arachidonate from PC to PE pools. Further evidence of a decrease in AA remodeling was shown by a significant decrease in microsomal CoA-independent transacylase activity. Levels of lyso-PC and lyso-PE were not altered in cells undergoing apoptosis, suggesting that the accumulation of lysophospholipids did not account for the decrease in CoA independent transacylase activity or the induction of apoptosis. Together, these data suggest that the mole quantities of free AA closely correlated with apoptosis and that the accumulation of AA in BMMC during apoptosis was mediated by a decreased capacity of these cells to remodel AA from PC to PE. PMID- 11022039 TI - Reconstitution of pterin-free inducible nitric-oxide synthase. AB - Inducible nitric-oxide synthase (NOS) was expressed and purified in the absence of 6(R)-tetrahydro-l-biopterin (H(4)B). Pterin-free NOS exhibits a Soret band (416-420 nm) characteristic of predominantly low spin heme and does not catalyze the formation of nitric oxide (. NO) (Rusche, K. M., Spiering, M. M., and Marletta, M. A. (1998) Biochemistry 37, 15503-15512). Reconstitution of pterin free NOS with H(4)B was monitored by a shift in the Soret band to 396-400 nm, the recovery of.NO-forming activity, and the measurement of H(4)B bound to the enzyme. As assessed by these properties, H(4)B binding was not rapid and required the presence of a reduced thiol. Spectral changes and recovery of activity were incomplete in the absence of reduced thiol. Full reconstitution of holoenzyme activity and stoichiometric H(4)B binding was achieved in the presence of 5 mm glutathione (GSH). Preincubation with GSH before the addition of H(4)B decreased, whereas lower concentrations of GSH extended, the time required for reconstitution. Six protected cysteine residues in pterin-free NOS were identified by labeling of NOS with cysteine-directed reagents before and after reduction with GSH. Heme and metal content of pterin-free and H(4)B-reconstituted NOS were also measured and were found to be independent of H(4)B content. Additionally, pterin-free NOS was reconstituted with 6-methylpterin analogs, including redox-stable deazapterins. Reconstitution with the redox-stable pterin analogs was neither time- nor thiol-dependent. Apparent binding constants were determined for the 6-methyl- (50 microm) and 6-ethoxymethyl (200 microm) deazapterins. The redox-stable pterin analogs appear to bind to NOS in a different manner than H(4)B. PMID- 11022040 TI - Serine residue in the IIIS5-S6 linker of the L-type Ca2+ channel alpha 1C subunit is the critical determinant of the action of dihydropyridine Ca2+ channel agonists. AB - The dihydropyridine (DHP)-binding site has been identified within L-type Ca(2+) channel alpha(1C) subunit. However, the molecular mechanism underlying modulation of Ca(2+) channel gating by DHPs has not been clarified. To search for novel determinants of high affinity DHP binding, we introduced point mutations in the rat brain Ca(2+) channel alpha(1C) subunit (rbCII or Ca(v)1.2c) based on the comparison of amino acid sequences between rbCII and the ascidian L-type Ca(2+) channel alpha(1) subunit, which is insensitive to DHPs. The alpha(1C) mutants (S1115A, S1146A, and A1420S) and rbCII were transiently expressed in BHK6 cells with beta(1a) and alpha(2)/delta subunits. The mutation did not affect the electrophysiological properties of the Ca(2+) channel, or the voltage- and concentration-dependent block of Ca(2+) channel currents produced by diltiazem and verapamil. However, the S1115A channel was significantly less sensitive to DHP antagonists. Interestingly, in the S1115A channel, DHP agonists failed to enhance whole-cell Ca(2+) channel currents and the prolongation of mean open time, as well as the increment of NP(o). Responsiveness to the non-DHP agonist FPL-64176 was also markedly reduced in the S1115A channel. When S1115 was replaced by other amino acids (S1115D, S1115T, or S1115V), only S1115T was slightly sensitive to S-(-)-Bay K 8644. These results indicate that the hydroxyl group of Ser(1115) in IIIS5-S6 linker of the L-type Ca(2+) channel alpha(1C) subunit plays a critical role in DHP binding and in the action of DHP Ca(2+) channel agonists. PMID- 11022041 TI - Residues in the 1A rod domain segment and the linker L2 are required for stabilizing the A11 molecular alignment mode in keratin intermediate filaments. AB - Both analyses of x-ray diffraction patterns of well oriented specimens of trichocyte keratin intermediate filaments (IF) and in vitro cross-linking experiments on several types of IF have documented that there are three modes of alignment of pairs of antiparallel molecules in all IF: A11, A22 and A12, based on which parts of the major rod domain segments are overlapped. Here we have examined which residues may be important for stabilizing the A11 mode. Using the K5/K14 system, we have made point mutations of charged residues along the chains and examined the propensities of equimolar mixtures of wild type and mutant chains to reassemble using as criteria: the formation (or not) of IF in vitro or in vivo; and stabilities of one- and two-molecule assemblies. We identified that the conserved residue Arg10 of the 1A rod domain, and the conserved residues Glu4 and Glu6 of the linker L2, were essential for stability. Additionally, conserved residues Lys31 of 1A and Asp1 of 2A and non-conserved residues Asp/Asn9 of 1A, Asp/Asn3 of 2A, and Asp7 of L2 are important for stability. Notably, these groups of residues lie close to each other when two antiparallel molecules are aligned in the A11 mode, and are located toward the ends of the overlap region. Although other sets of residues might theoretically also contribute, we conclude that these residues in particular engage in favorable intermolecular ionic and/or H bonding interactions and thereby may play a role in stabilizing the A11 mode of alignment in keratin IF. PMID- 11022042 TI - Association of COOH-terminal-binding protein (CtBP) and MEF2-interacting transcription repressor (MITR) contributes to transcriptional repression of the MEF2 transcription factor. AB - The class II histone deacetylases (HDACs) 4, 5, and 7 share a common structural organization, with a carboxyl-terminal catalytic domain and an amino-terminal extension that mediates interactions with members of the myocyte enhancer factor 2 (MEF2) family of transcription factors. Association of these HDACs with MEF2 factors represses transcription of MEF2 target genes. MEF2-interacting transcription repressor (MITR) shares homology with the amino-terminal extensions of class II HDACs and also acts as a transcriptional repressor, but lacks a histone deacetylase catalytic domain. This suggests that MITR represses transcription by recruiting other corepressors. We show that the amino-terminal regions of MITR and class II HDACs interact with the transcriptional corepressor, COOH-terminal-binding protein (CtBP), through a CtBP-binding motif (P-X-D-L-R) conserved in MITR and HDACs 4, 5, and 7. Mutation of this sequence in MITR abolishes interaction with CtBP and impairs, but does not eliminate, the ability of MITR to inhibit MEF2-dependent transcription. The residual repressive activity of MITR mutants that fail to bind CtBP can be attributed to association with other HDAC family members. These findings reveal CtBP-dependent and -independent mechanisms for transcriptional repression by MITR and show that MITR represses MEF2 activity through recruitment of multicomponent corepressor complexes that include CtBP and HDACs. PMID- 11022043 TI - Mutually cooperative binding of eukaryotic translation initiation factor (eIF) 3 and eIF4A to human eIF4G-1. AB - Eukaryotic translation initiation factor 4G-1 (eIF4G) plays a critical role in the recruitment of mRNA to the 43 S preinitiation complex. The central region of eIF4G binds the ATP-dependent RNA helicase eIF4A, the 40 S binding factor eIF3, and RNA. In the present work, we have further characterized the binding properties of the central region of human eIF4G. Both titration and competition experiments were consistent with a 1:1 stoichiometry for eIF3 binding. Surface plasmon resonance studies showed that three recombinant eIF4G fragments corresponding to amino acids 642-1560, 613-1078, and 975-1078 bound eIF3 with similar kinetics. A dissociation equilibrium constant of approximately 42 nm was derived from an association rate constant of 3.9 x 10(4) m(-1) s(-1) and dissociation rate constant of 1.5 x 10(-3) s(-1). Thus, the eIF3-binding region is included within amino acid residues 975-1078. This region does not overlap with the RNA-binding site, which suggests that eIF3 binds eIF4G directly and not through an RNA bridge, or the central eIF4A-binding site. Surprisingly, the binding of eIF3 and eIF4A to the central region was mutually cooperative; eIF3 binding to eIF4G increased 4-fold in the presence of eIF4A, and conversely, eIF4A binding to the central (but not COOH-terminal) region of eIF4G increased 2.4-fold in the presence of eIF3. PMID- 11022044 TI - Identification of overlapping AP-2/NF-kappa B-responsive elements on the rat cholecystokinin gene promoter. AB - In this study we evaluate both proximal and more distal transcriptional regulation of the 5' flanking region of the rat cholecystokinin gene in transfected GH3 (rat pituitary tumor) cells. Transcriptional activity was measured on the intact (-400 to +73) 5' flanking region of cholecystokinin (CCK), as well as with DNA constructs, which were deleted in both the conventional 5' to 3', as well as an unconventional 3' to 5' direction. Our in vivo studies indicate complex phorbol ester and forskolin interactions in the 10-base pair region between -130 and -140. We conclude, there are at least two transcriptional factors involved in regulation of the rat CCK transcription in this region. In vitro studies utilizing heterologous nuclear (HeLa) extract, as well as purified transcription factors AP-2 and NF-kappa B, identify overlapped AP-2- and NF-kappa B-responsive elements within the 17-base pair sequence between -149 and -134 of the distal 5' flanking region. In this region complex transcriptional regulation occurs, which indicates inhibition of AP-2 CCK promoter complexing by NF-kappa B. Six-point mutations introduced into this sequence prevent AP-2 and NF-kappa B binding to CCK promoter, as well as its transcriptional activation by phorbol ester and forskolin in GH3 cells. PMID- 11022045 TI - Co-translational interactions of apoprotein B with the ribosome and translocon during lipoprotein assembly or targeting to the proteasome. AB - Hepatic lipoprotein assembly and secretion can be regulated by proteasomal degradation of newly synthesized apoB, especially if lipid synthesis or lipid transfer is low. Our previous studies in HepG2 cells showed that, under these conditions, newly synthesized apoB remains stably associated with the endoplasmic reticulum (ER) membrane (Mitchell, D. M., Zhou, M., Pariyarath, R., Wang, H., Aitchison, J. D., Ginsberg, H. N., and Fisher, E. A. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 14733-14738). We now show that independent of lipid synthesis, apoB chains that appear full-length are, in fact, incompletely translated polypeptides still engaged by the ribosome and associated with the ER translocon. In the presence of active lipid synthesis and transfer, translation and lipoprotein assembly are completed, and the complexes exit the ER. Upon omitting fatty acids from, or adding a microsomal triglyceride transfer protein inhibitor to, culture media to reduce lipid synthesis or transfer, respectively, apoB was degraded while it remained associated with the ER and complexed with cytosolic hsp70 and proteasomes. Thus, unlike other ER substrates of the proteasome, such as major histocompatibility complex class I molecules, apoB does not fully retrotranslocate to the cytosol before entering the ubiquitin-proteasome pathway. Although, upon immunofluorescence, apoB in proteasome-inhibited cells accumulated in punctate structures similar in appearance to aggresomes (cytosolic structures containing molecules irreversibly lost from the secretory pathway), these apoB molecules could be secreted when lipid synthesis was stimulated. The results suggest a model in which 1) apoB translation does not complete until lipoprotein assembly terminates, and 2) assembly with lipids or entry into the ubiquitin proteasome pathway occurs while apoB polypeptides remain associated with the translocon and attached to the ribosome. PMID- 11022046 TI - Epithelial sodium channel pore region. structure and role in gating. AB - Epithelial sodium channels (ENaC) have a crucial role in the regulation of extracellular fluid volume and blood pressure. To study the structure of the pore region of ENaC, the susceptibility of introduced cysteine residues to sulfhydryl reactive methanethiosulfonate derivatives ((2-aminoethyl)methanethiosulfonate hydrobromide (MTSEA) and [(2-(trimethylammonium)ethyl]methanethiosulfonate bromide (MTSET)) and to Cd(2+) was determined. Selected mutants within the amino terminal portion (alphaVal(569)-alphaTrp(582)) of the pore region responded to MTSEA, MTSET, or Cd(2+) with stimulation or inhibition of whole cell Na(+) current. The reactive residues were not contiguous but were separated by 2-3 residues where substituted cysteine residues did not respond to the reagents and line one face of an alpha-helix. The activation of alphaS580Cbetagamma mENaC by MTSET was associated with a large increase in channel open probability. Within the carboxyl-terminal portion (alphaSer(583)-alphaSer(592)) of the pore region, only one mutation (alphaS583C) conferred a rapid, nearly complete block by MTSEA, MTSET, and Cd(2+), whereas several other mutant channels were partially blocked by MTSEA or Cd(2+) but not by MTSET. Our data suggest that the outer pore of ENaC is formed by an alpha-helix, followed by an extended region that forms a selectivity filter. Furthermore, our data suggest that the pore region participates in ENaC gating. PMID- 11022047 TI - A novel bifunctional phospholipase c that is regulated by Galpha 12 and stimulates the Ras/mitogen-activated protein kinase pathway. AB - Three families of phospholipase C (PI-PLCbeta, gamma, and delta) are known to catalyze the hydrolysis of polyphosphoinositides such as phosphatidylinositol 4,5 bisphosphate (PIP(2)) to generate the second messengers inositol 1,4,5 trisphosphate and diacylglycerol, leading to a cascade of intracellular responses that result in cell growth, cell differentiation, and gene expression. Here we describe the founding member of a novel, structurally distinct fourth family of PI-PLC. PLCepsilon not only contains conserved catalytic (X and Y) and regulatory domains (C2) common to other eukaryotic PLCs, but also contains two Ras associating (RA) domains and a Ras guanine nucleotide exchange factor (RasGEF) motif. PLCepsilon hydrolyzes PIP(2), and this activity is stimulated selectively by a constitutively active form of the heterotrimeric G protein Galpha(12). PLCepsilon and a mutant (H1144L) incapable of hydrolyzing phosphoinositides promote formation of GTP-Ras. Thus PLCepsilon is a RasGEF. PLCepsilon, the mutant H1144L, and the isolated GEF domain activate the mitogen-activated protein kinase pathway in a manner dependent on Ras but independent of PIP(2) hydrolysis. Our findings demonstrate that PLCepsilon is a novel bifunctional enzyme that is regulated by the heterotrimeric G protein Galpha(12) and activates the small G protein Ras/mitogen-activated protein kinase signaling pathway. PMID- 11022048 TI - Regulation of a novel human phospholipase C, PLCepsilon, through membrane targeting by Ras. AB - Phosphoinositide-specific phospholipase C (PI-PLC) plays a pivotal role in regulation of intracellular signal transduction from various receptor molecules. More than 10 members of human PI-PLC isoforms have been identified and classified into three classes beta, gamma, and delta, which are regulated by distinct mechanisms. Here we report identification of a novel class of human PI-PLC, named PLCepsilon, which is characterized by the presence of a Ras-associating domain at its C terminus and a CDC25-like domain at its N terminus. The Ras-associating domain of PLCepsilon specifically binds to the GTP-bound forms of Ha-Ras and Rap1A. The dissociation constant for Ha-Ras is estimated to be approximately 40 nm, comparable with those of other Ras effectors. Co-expression of an activated Ha-Ras mutant with PLCepsilon induces its translocation from the cytosol to the plasma membrane. Upon stimulation with epidermal growth factor, similar translocation of ectopically expressed PLCepsilon is observed, which is inhibited by co-expression of dominant-negative Ha-Ras. Furthermore, using a liposome-based reconstitution assay, it is shown that the phosphatidylinositol 4,5-bisphosphate hydrolyzing activity of PLCepsilon is stimulated in vitro by Ha-Ras in a GTP dependent manner. These results indicate that Ras directly regulates phosphoinositide breakdown through membrane targeting of PLCepsilon. PMID- 11022049 TI - Differential contributions of NADPH-cytochrome P450 oxidoreductase FAD binding site residues to flavin binding and catalysis. AB - Transfer of reducing equivalents from NADPH to the cytochromes P450 is mediated by NADPH-cytochrome P450 oxidoreductase, which contains stoichiometric amounts of tightly bound FMN and FAD. Hydrogen bonding and van der Waals interactions between FAD and amino acid residues in the FAD binding site of the reductase serve to regulate both flavin binding and reactivity. The precise orientation of key residues (Arg(454), Tyr(456), Cys(472), Gly(488), Thr(491), and Trp(677)) has been defined by x-ray crystallography (Wang, M., Roberts, D. L., Paschke, R., Shea, T. M., Masters, B. S., Kim, J.-J. P. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 8411-8416). The current study examines the relative contributions of these residues to FAD binding and catalysis by site-directed mutagenesis and kinetic analysis. Mutation of either Tyr(456), which makes van der Waals contact with the FAD isoalloxazine ring and also hydrogen-bonds to the ribityl 4'-hydroxyl, or Arg(454), which bonds to the FAD pyrophosphate, decreases the affinity for FAD 8000- and 25,000-fold, respectively, with corresponding decreases in cytochrome c reductase activity. In contrast, substitution of Thr(491), which also interacts with the pyrophosphate grouping, had a relatively modest effect on both FAD binding (100-fold decrease) and catalytic activity (2-fold decrease), while the G488L mutant exhibited, respectively, 800- and 50-fold decreases in FAD binding and catalytic activity. Enzymic activity of each of these mutants could be restored by addition of FAD. Kinetic properties and the FMN content of these mutants were not affected by these substitutions, with the exception of a 3-fold increase in Y456S K(m)(cyt )(c) and a 70% decrease in R454E FMN content, suggesting that the FMN- and FAD-binding domains are largely, but not completely, independent. Even though Trp(677) is stacked against the re-face of FAD, suggesting an important role in FAD binding, deletion of both Trp(677) and the carboxyl-terminal Ser(678) decreased catalytic activity 50-fold without affecting FAD content. PMID- 11022050 TI - Intersubunit interactions in Plasmodium falciparum thioredoxin reductase. AB - The thioredoxin redox system is composed of the NADPH-dependent homodimeric flavoprotein thioredoxin reductase (TrxR) and the 12-kDa protein thioredoxin. It is responsible for the reduction of disulfide bridges in proteins such as ribonucleotide reductase and several transcription factors. Furthermore, thioredoxin is involved in the detoxification of hydrogen peroxide and protects the cell against oxidative damage. There exist two classes of TrxRs: the high M(r) and the low M(r) proteins. The well characterized Escherichia coli TrxR represents a member of the low M(r) class of proteins, whereas the mammalian, Caenorhabditis elegans, and Plasmodium falciparum proteins belong to the family of high M(r) proteins. The primary structure of these proteins is very similar to that of glutathione reductase and lipoamide dehydrogenase. However, the high M(r) TrxRs possess, in addition to their redox active N-terminal pair of cysteines, a pair of cysteine residues or a selenenylsulfide motif at their C terminus. These residues have been shown to be crucial for the reduction of thioredoxin. In this study we address the question whether the active site residues of P. falciparum TrxR are provided by one or both subunits. Differentially tagged wild-type and PfTrxR mutants were co-expressed in E. coli and the recombinant protein species were purified by affinity chromatography specific for the respective tags of the recombinant proteins. Co-expression of PfTrxR wild-type and mutant proteins resulted in the formation of three different protein species: homodimeric PfTrxR wild-type proteins, homodimeric mutant proteins, and heterodimers composed of one PfTrxR wild-type subunit and one PfTrxR mutant subunit. Co-expression of the double mutant PfTrxRC88AC535A with PfTrxR wild-type generated an inactive heterodimer, which indicates that PfTrxR possesses intersubunit active sites. In addition, the data presented possibly imply a coopertive interaction between both active sites of PfTrxR. PMID- 11022052 TI - Serum vitamin C concentration was inversely associated with subsequent 20-year incidence of stroke in a Japanese rural community. The Shibata study. AB - BACKGROUND AND PURPOSE: Epidemiological evidence suggests that vitamin C may decrease the risk of stroke. The purpose of the present study was to examine the association of serum vitamin C concentration with the subsequent incidence of stroke. METHODS: In a Japanese rural community, a cohort of 880 men and 1241 women aged 40 years and older who were initially free of stroke was examined in 1977 and followed until 1997. The baseline examination included a measurement of serum vitamin C concentration. The incidence of stroke was determined by annual follow-up examinations and registry. RESULTS: During the 20-year observation period, 196 incident cases of all stroke, including 109 cerebral infarctions and 54 hemorrhagic strokes, were documented. Strong inverse associations were observed between serum vitamin C concentration and all stroke (sex- and age adjusted hazard ratios were 0.93, 0.72, and 0.59, respectively, for the second, third, and fourth quartiles compared with the first quartile; P for trend=0.002), cerebral infarction (0.71, 0.59, and 0.51; P for trend=0.015), and hemorrhagic stroke (0.89, 0.75, and 0. 45; P for trend=0.013). Additional adjustments for blood pressure, serum total cholesterol, body mass index, physical activity, smoking, alcohol drinking, antihypertensive medication, atrial fibrillation, and history of ischemic heart disease did not attenuate these associations markedly. CONCLUSIONS: Serum vitamin C concentration was inversely related to the subsequent incidence of stroke. This relationship was significant for both cerebral infarction and hemorrhagic stroke. Additional mechanistic hypotheses may be required to explain our findings. PMID- 11022051 TI - The transcriptional and DNA binding activity of peroxisome proliferator-activated receptor alpha is inhibited by ethanol metabolism. A novel mechanism for the development of ethanol-induced fatty liver. AB - Fatty acids are ligands for the peroxisome proliferator-activated receptor alpha (PPAR alpha). Fatty acid levels are increased in liver during the metabolism of ethanol and might be expected to activate PPAR alpha. However, ethanol inhibited PPAR alpha activation of a reporter gene in H4IIEC3 hepatoma cells expressing alcohol-metabolizing enzymes but not in CV-1 cells, which lack these enzymes. Ethanol also reduced the ability of the PPAR alpha ligand WY14,643 to activate reporter constructs in the hepatoma cells or cultured rat hepatocytes. This effect of ethanol was abolished by the alcohol dehydrogenase inhibitor 4 methylpyrazole and augmented by the aldehyde dehydrogenase inhibitor cyanamide, indicating that acetaldehyde was responsible for the action of ethanol. PPAR alpha/retinoid X receptor extracted from hepatoma cells exposed to ethanol or acetaldehyde bound poorly to an oligonucleotide containing peroxisome proliferator response elements. This effect was also blocked by 4-methylpyrazole and augmented by cyanamide. Furthermore, in vitro translated PPAR alpha exposed to acetaldehyde failed to bind DNA. Thus, ethanol metabolism blocks transcriptional activation by PPAR alpha, in part due to impairment of its ability to bind DNA. This effect of ethanol may promote the development of alcoholic fatty liver and other hepatic consequences of alcohol abuse. PMID- 11022053 TI - Antioxidant profile and early outcome in stroke patients. AB - BACKGROUND AND PURPOSE: Experimental studies provide evidence of an association between ischemic stroke and increased oxidative stress, but data in humans are still limited and controversial. The purpose of this study was to investigate the time course of plasma antioxidant changes in ischemic stroke patients. METHODS: Plasma antioxidants, including water-soluble (vitamin C and uric acid) and lipid soluble (vitamins A and E) compounds as well as antioxidant enzyme activities in plasma (superoxide dismutase [SOD] and glutathione peroxidase) and erythrocytes (SOD), were measured by high-performance liquid chromatography (antioxidant vitamins) and by spectrophotometry (antioxidant enzymes) in 38 subjects (25 men and 13 women aged 77.2+/-7.9 years) with acute ischemic stroke of recent onset (<24 hours) on admission, after 6 and 24 hours, and on days 3, 5, and 7. Antioxidant levels in patients on admission were compared with those of age- and sex-matched controls. RESULTS: Mean antioxidant levels and activities in patients on admission were lower than those of controls and showed a gradual increase over time. Patients with the worst early outcome (death or functional decline) had higher vitamin A and uric acid plasma levels and lower vitamin C levels and erythrocyte SOD activity than those who remained functionally stable. CONCLUSIONS: These results suggest that the majority of antioxidants are reduced immediately after an acute ischemic stroke, possibly as a consequence of increased oxidative stress. A specific antioxidant profile is associated with a poor early outcome. PMID- 11022054 TI - Intake of flavonoids, carotenoids, vitamins C and E, and risk of stroke in male smokers. AB - BACKGROUND AND PURPOSE: Antioxidants may protect against atherosclerosis and thus prevent cerebrovascular disease. We studied the association between dietary antioxidants and subtypes of stroke. METHODS: The study cohort consisted of 26 593 male smokers, aged 50 to 69 years, without a history of stroke. They were participants of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study in Finland. The men completed a validated dietary questionnaire at baseline. Incident cases were identified through national registers. RESULTS: During a 6.1-year follow-up, 736 cerebral infarctions, 83 subarachnoid hemorrhages, and 95 intracerebral hemorrhages occurred. Neither dietary flavonols and flavones nor vitamin E were associated with risk for stroke. The dietary intake of beta-carotene was inversely associated with the risk for cerebral infarction (relative risk [RR] of highest versus lowest quartile 0.74, 95% CI 0.60 to 0. 91), lutein plus zeaxanthin with risk for subarachnoid hemorrhage (RR 0.47, 95% CI 0.24 to 0.93), and lycopene with risks of cerebral infarction (RR 0.74, 95% CI 0.59 to 0.92) and intracerebral hemorrhage (RR 0.45, 95% CI 0.24 to 0.86). Vitamin C intake was inversely associated with the risk for intracerebral hemorrhage (RR 0.39, 95% CI 0.21 to 0.74). After simultaneous modeling of the antioxidants, a significant association remained only between beta-carotene intake and risk for cerebral infarction (RR 0.77, 95% CI 0.61 to 0.99). CONCLUSIONS: Dietary intake of beta-carotene was inversely associated with the risk for cerebral infarction. No association was detected between other dietary antioxidants and risk for stroke. PMID- 11022055 TI - Orthostatic hypotension as a risk factor for stroke: the atherosclerosis risk in communities (ARIC) study, 1987-1996. AB - BACKGROUND AND PURPOSE: The association between orthostatic hypotension (OH) and stroke has rarely been investigated in longitudinal studies. The purpose of the present study was to determine whether OH predicts ischemic stroke in a middle aged, biethnic population after adjustment for known stroke risk factors. Diastolic, systolic, and consensus OH were evaluated for baseline associations and for the ability to predict stroke. METHODS: In 11 707 persons from the Atherosclerosis Risk in Communities (ARIC) cohort who were free of stroke and overt heart disease at baseline, Cox proportional hazards analyses modeled the association between OH at baseline and incident ischemic stroke over 7.9 years of follow-up. OH was defined as a systolic blood pressure drop >/=20 mm Hg (systolic OH), a diastolic blood pressure drop >/=10 mm Hg (diastolic OH), or a drop in either (consensus OH) when a person changed from a supine to standing position. RESULTS: OH was predictive of ischemic stroke, even after adjustment for numerous stroke risk factors (consensus OH: hazard ratio, 2.0; 95% CI, 1.2 to 3.2). While the baseline characteristics associated with OH varied depending on the type of OH, all types of OH had a similar risk of stroke. CONCLUSIONS: OH is an easily obtained measurement that may help to identify middle-aged persons at risk for stroke. PMID- 11022056 TI - Cerebral autoregulation in subjects adapted and not adapted to high altitude. AB - BACKGROUND AND PURPOSE: Impaired cerebral autoregulation (CA) from high-altitude hypoxia may cause high-altitude cerebral edema in newcomers to a higher altitude. Furthermore, it is assumed that high-altitude natives have preserved CA. However, cerebral autoregulation has not been studied at altitude. METHODS: We studied CA in 10 subjects at sea level and in 9 Sherpas and 10 newcomers at an altitude of 4243 m by evaluating the effect of an increase of mean arterial blood pressure (MABP) with phenylephrine infusion on the blood flow velocity in the middle cerebral artery (Vmca), using transcranial Doppler. Theoretically, no change of Vmca in response to an increase in MABP would imply perfect autoregulation. Complete loss of autoregulation is present if Vmca changes proportionally with changes of MABP. RESULTS: In the sea-level group, at a relative MABP increase of 23+/-4% during phenylephrine infusion, relative Vmca did not change essentially from baseline Vmca (2+/-7%, P=0.36), which indicated intact autoregulation. In the Sherpa group, at a relative MABP increase of 29+/-7%, there was a uniform and significant increase of Vmca of 24+/-9% (P<0.0001) from baseline Vmca, which indicated loss of autoregulation. The newcomers showed large variations of Vmca in response to a relative MABP increase of 21+/-6%. Five subjects showed increases of Vmca of 22% to 35%, and 2 subjects showed decreases of Vmca of 21% and 23%. CONCLUSIONS: All Sherpas and the majority of the newcomers showed impaired CA. It indicates that an intact autoregulatory response to changes in blood pressure is probably not a hallmark of the normal human cerebral vasculature at altitude and that impaired CA does not play a major role in the occurrence of cerebral edema in newcomers to the altitude. PMID- 11022057 TI - Association between risk factors for atherosclerosis and mechanical forces in carotid artery. AB - BACKGROUND AND PURPOSE: Mechanical stresses on the arterial wall participate in the pathogenesis of atherosclerosis as local factors. The relationships between local mechanical forces and risk factors for atherosclerosis were investigated. METHODS: Mechanical forces on the arterial wall were evaluated in the carotid artery in 117 patients with risk factors for atherosclerosis including hypertension, dyslipidemia, diabetes mellitus, and smoking, as well as in 20 age- and sex-matched normal controls. Circumferential wall tension and shear stress were evaluated with Laplace's law and a poiseuillean parabolic model of velocity distribution. Circumferential wall strain was also evaluated as carotid mechanical force. RESULTS: Mechanical forces in subjects with risk factors were characterized by low wall shear stress, high circumferential wall tension, and reduced strain. Systolic blood pressure was significantly negatively associated with shear stress and circumferential wall strain. HDL cholesterol showed a significant positive correlation with shear stress and a negative correlation with wall tension. Fasting blood glucose was significantly associated with shear stress, while smoking showed a negative correlation with shear stress and a positive correlation with wall tension. Accumulation of risk factors was associated with further deterioration of mechanical forces. Furthermore, stepwise regression analysis showed that the number of risk factors was significantly associated with mechanical forces independently of carotid intima-media thickness. CONCLUSIONS: These findings suggest that risk factors for atherosclerosis were associated with alteration of mechanical forces. Consequent alteration in mechanical forces could be an underlying local mechanism for the progression of atherosclerosis. PMID- 11022058 TI - Proinflammatory cytokines and early neurological worsening in ischemic stroke. AB - BACKGROUND AND PURPOSE: The mechanisms for clinical deterioration in patients with ischemic stroke are not completely understood. Several proinflammatory cytokines are released early after the onset of brain ischemia, but it is unknown whether inflammation predisposes to neurological deterioration. We assessed the implication of interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha in early neurological worsening in ischemic stroke. METHODS: Two hundred thirty-one patients consecutively admitted with first-ever ischemic cerebral infarction within the first 24 hours from onset were included. Neurological worsening was defined when the Canadian Stroke Scale (CSS) score fell at least 1 point during the first 48 hours after admission. IL-6 and TNF-alpha were determined in plasma and cerebrospinal fluid (CSF; n=81) obtained on admission. RESULTS: Eighty-three patients (35.9%) deteriorated within the first 48 hours. IL-6 in plasma (>21.5 pg/mL; OR 37.7, CI 11.9 to 118.8) or in CSF (>6.3 pg/mL; OR 13.1, CI 2.2 to 77.3) were independent factors for early clinical worsening, with multiple logistic regression. The association was statistically significant in all ischemic stroke subtypes as well as in subjects with cortical or subcortical infarctions. IL-6 in plasma was highly correlated with body temperature, glucose, fibrinogen, and infarct volume. CSF and plasma concentrations of TNF-alpha were also higher in patients who deteriorated, but the differences observed did not remain significant on multivariate analysis. CONCLUSIONS: In addition to participating in the acute-phase response that follows focal cerebral ischemia, IL-6 levels on admission are associated with early clinical deterioration. The association between IL-6 and early neurological worsening prevails without regard to the initial size, topography, or mechanism of the ischemic infarction. PMID- 11022059 TI - Effect of contralateral occlusion on long-term efficacy of endarterectomy in the asymptomatic carotid atherosclerosis study (ACAS). ACAS Investigators. AB - BACKGROUND AND PURPOSE: The Asymptomatic Carotid Atherosclerosis Study (ACAS) established the effectiveness of prophylactic carotid endarterectomy, for patients in good health who had stenosis >/=60%, if conducted by surgeons with a surgical morbidity and mortality of <3%. This secondary analysis was performed to determine whether the presence of contralateral cervical carotid occlusion alters the efficacy of asymptomatic ipsilateral carotid endarterectomy. METHODS: One hundred sixty-three participants who had a baseline contralateral occlusion documented by Doppler ultrasound (77 medical, 86 surgical) were compared with 1485 participants with a patent contralateral carotid artery (748 medical, 737 surgical) for the risk of a combined end point of perioperative (30-day) death or stroke or long-term (5-year) ipsilateral stroke. RESULTS: For those without contralateral occlusion, surgery was associated with a 6.7% absolute reduction in the 5-year risk (95% CI, 2.1% to 11.4%), while for those with a contralateral occlusion, surgery was associated with a 2.0% absolute increase in risk (95% CI, 9.3% to 5.2%), which was a statistically significant difference in the effect of surgery (P:=0.047). This difference is primarily attributable to low long-term risk for medically managed patients with contralateral occlusion. CONCLUSIONS: While this post hoc analysis should be interpreted with caution, the findings suggest that endarterectomy in asymptomatic subjects with contralateral occlusion provides no long-term benefit (and may be harmful) in preventing stroke and death. These findings were a result of the benign course of medically treated subjects. PMID- 11022060 TI - Finding the most powerful measures of the effectiveness of tissue plasminogen activator in the NINDS tPA stroke trial. AB - BACKGROUND AND PURPOSE: We sought to identify the most powerful binary measures of the treatment effect of tissue plasminogen activator (tPA) in the National Institute of Neurological Disorders and Stroke (NINDS) rTPA Stroke Trial. METHODS: Using the Classification and Regression Tree (CART) algorithm, we evaluated binary cut points and combination of binary cut points with the 4 clinical scales and head CT imaging measures in the NINDS tPA Stroke Trial at 4 times after treatment: 2 hours, 24 hours, 7 to 10 days, and 3 months. The first analysis focused on detecting evidence of "early activity" of tPA with the use of outcome measures derived from the 2-hour and 24-hour clinical and radiographic measures. The second analysis focused on longer-term outcome and "efficacy" and used outcome measures derived from 7- to 10-day and 3-month measures. After identifying the cut points with the ability to classify patients into the tPA and placebo groups using part I data from the trial, we then used data from part II of the trial to validate the results. RESULTS: Of the 5 most powerful outcome measures for early activity of tPA, 4 involved the National Institutes of Health Stroke Scale (NIHSS) score at 24 hours or changes in the NIHSS score from baseline to 24 hours. The best overall single outcome measure was an NIHSS score /=65 years on December 31, 1988, whose strokes occurred in the period 1989-1993, were followed for mortality until July 1, 1995. Personal data were gathered through home interviews conducted by well-trained community nurses, and mortality data were obtained from the national census office by using identification card numbers. Cox proportional hazards regression analysis and the stepwise technique were used to search for important prognostic factors of survival. RESULTS: Women experienced a higher mortality rate (139.8 per 1000 person-years) than men (126.4 per 1000 person-years), as age-adjusted for World Health Organization world-population figures. Stroke patients who received continuous treatment for diabetes experienced mortality risks similar to those of patients without diabetes and much lower risks than those with discontinuous diabetes treatment. Cognitive impairment was also an independent predictor of survival (relative risk 2.69, P<0.05). In addition, patients with both cognitive and mobility impairments had a 2- to 3-fold greater risk of mortality than those with only a single abnormality. CONCLUSIONS: This first report on the various prognostic factors related to survival of elderly stroke patients in Taiwan's Chinese population emphasized the benefit of continuous diabetes treatment in improving survival chances. These stroke patients should also be monitored for cognitive and mobility impairments and undergo rehabilitation. PMID- 11022064 TI - Determinants of neurological outcome after surgery for brain arteriovenous malformation. AB - BACKGROUND AND PURPOSE: We sought to define determinants of neurological deficit after surgery for brain arteriovenous malformation (AVM). METHODS: One hundred twenty-four prospective patients (48% women, mean age 33 years) underwent microsurgical brain AVM resection. Patients were examined by 3 study neurologists immediately before surgery, postoperatively in-hospital, by in-person long-term follow-up, and with a structured telephone follow-up. They were classified according to the 5-point Spetzler-Martin grading system, with its 3 elements: size, venous drainage pattern, and location. The functional neurological status was classified with the modified Rankin scale. Multivariate logistic regression models were applied to test the effect of patient age, gender, and the 3 Spetzler Martin elements on early and long-term postoperative neurological complications. RESULTS: Twelve patients (10%) were classified as Spetzler-Martin grade 1; 36 (29%) as grade 2; 47 (38%) as grade 3; 26 (21%) as grade 4; and 3 (2%) as grade 5. Postoperatively, in-hospital, 51 patients (41%) showed new neurological deficits (15% disabling [ie, Rankin scale score >2] and 26% nondisabling [ie, Rankin 1 or 2]). At long-term follow-up (mean follow-up time 12 months), 47 patients (38%) revealed surgery-related neurological deficits (6% disabling; 32% nondisabling). The rate of neurological complications increased by Spetzler Martin grade. Female gender, AVM size, and deep venous drainage were significantly associated with neurological deficits at in-hospital and long-term evaluation. For patient age and AVM location, no significant association was found. CONCLUSIONS: The findings suggest that female gender, AVM size, and AVM drainage into the deep venous system may be determinants of neurological deficit after microsurgical AVM resection. PMID- 11022065 TI - Incident hemorrhage risk of brain arteriovenous malformations located in the arterial borderzones. AB - BACKGROUND AND PURPOSE: We sought to assess the relative risk of hemorrhagic presentation of brain arteriovenous malformations (AVMs) located in the arterial borderzone territories. METHODS: The 464 consecutive, prospectively enrolled patients from the New York AVM Databank were analyzed. AVM borderzone location was coded positive when the malformation was supplied by branches of at least 2 of the major circle of Willis arteries (anterior, middle, and/or posterior cerebral arteries). AVMs fed by branches of only 1 major pial or any other single artery served as a comparison group. Clinical presentation (diagnostic event) was categorized as (1) intracranial hemorrhage, proven by brain imaging, or (2) seizure, focal neurological deficit, headache, or other event with no signs of AVM hemorrhage on brain imaging. RESULTS: In 48% (n=222) of the patients, AVMs were located in the arterial borderzone territories; in 52% (n=242) a non borderzone location was found. Hemorrhage was the presenting symptom in 44% (n=205); 28% (n=132) presented with seizures, 11% (n=52) with headaches, 7% (n=34) with a neurological deficit, and 9% (n=41) with other or no AVM-related symptoms. The frequency of incident AVM hemorrhage was significantly lower in borderzone AVMs (27%, n=61) than in non-borderzone malformations (60%, n=144; P:<0.001). This difference remained significant in a multivariate model controlling for age, sex, AVM size, deep venous drainage, and presence of aneurysms (odds ratio, 0.4; 95% CI, 0.25 to 0.66). CONCLUSIONS: Our findings suggest that borderzone location is an independent determinant for a lower risk of AVM hemorrhage at initial presentation. PMID- 11022066 TI - Outcomes of early endovascular versus surgical treatment of ruptured cerebral aneurysms. A prospective randomized study. AB - BACKGROUND AND PURPOSE: This prospective study was conducted to compare the outcomes of surgical clipping and endovascular treatment in acute (<72 hours) aneurysmal subarachnoid hemorrhage (SAH). METHODS: One hundred nine consecutive patients were randomly assigned to either surgical (n=57) or endovascular (n=52) treatment. Clinical and neuropsychological outcome was assessed at 3 and 12 months after treatment; MRI of the brain was performed at 12 months. Follow-up angiography was scheduled after clipping and 3 and 12 months after endovascular treatment. RESULTS: One year postoperatively, 43/41 (surgical/endovascular) patients had good or moderate recovery, 5/4 had severe disability or were in a vegetative state, and 9/7 had died (NS) according to intention to treat. Patients with good clinical recovery did not differ in their neuropsychological test scores. Symptomatic vasospasm (OR 2.47; 95% CI 1.45 to 4.19; P<0.001), poorer Hunt and Hess grade (OR 2.50; 95% CI 1.31 to 4.75; P=0.005), need for permanent shunt (OR 8.90; 95% CI 1.80 to 44.15; P=0.008), and larger size of the aneurysm (OR 1. 22; 95% CI 1.02 to 1.45; P=0.032) independently predicted worsened clinical outcome regardless of the treatment modality. In MRI, superficial brain retraction deficits (P<0.001) and ischemic lesions in the territory of the ruptured aneurysm (P=0.025) were more frequent in the surgical group. Kaplan Meier analysis (mean+/-SD follow-up 39+/-18 months) revealed equal survival in both treatment groups. No late rebleedings have occurred. CONCLUSIONS: One-year clinical and neuropsychological outcomes seem comparable after early surgical and endovascular treatment of ruptured intracranial aneurysms. The long-term efficacy of endovascular treatment in preventing rebleeding remains open. PMID- 11022067 TI - Relationship between apparent diffusion coefficient and subsequent hemorrhagic transformation following acute ischemic stroke. AB - BACKGROUND AND PURPOSE: A method for identifying patients at increased risk for developing secondary hemorrhagic transformation (HT) after acute ischemic stroke could be of significant value, particularly in patients being considered for thrombolytic therapy. We hypothesized that diffusion-weighted MRI might aid in the identification of such patients. METHODS: We retrospectively analyzed 17 patients with ischemic stroke who received diffusion-weighted MRI within 8 hours of symptom onset and who also received follow-up neuroimaging within 1 week of initial scan. The apparent diffusion coefficient (ADC) for each pixel in the whole ischemic area was calculated, generating a histogram of values. Areas subsequently experiencing HT were then compared with areas not experiencing HT to determine the relationship between ADC and subsequent HT. RESULTS: A significantly greater percentage of pixels possessed lower ADCs (40% of the pixels possessed values 30 mg/dL) was found in 20 patients and 10 controls (OR/95% CI, 4.84/2. 16 to 10.86); FV G1691A was present in 17 patients and 10 controls (OR/95% CI, 3.95/1.72 to 9.0); the PT G20210A variant was detected in 4 patients and 4 controls (OR/95% CI, 2.04/0.49 to 8.3); the MTHFR TT677 genotype was found in 15 patients and 20 controls (OR/95% CI, 1.59/0.77 to 3.29); and protein C type I deficiency was found in 6 neonates. Neither antithrombin deficiency nor protein S deficiency was found in the neonatal patients studied. Acquired IgG ACAs were found in 3 cases. Additional triggering factors, ie, asphyxia, septicemia, maternal diabetes, and perinatally acquired renal venous thrombosis, were reported in 54.0% of patients. CONCLUSIONS: Besides acquired triggering factors, the data presented here suggest that genetic prothrombotic risk factors play a role in symptomatic neonatal stroke. PMID- 11022078 TI - Atorvastatin upregulates type III nitric oxide synthase in thrombocytes, decreases platelet activation, and protects from cerebral ischemia in normocholesterolemic mice. AB - BACKGROUND AND PURPOSE: Thrombosis superimposed on atherosclerosis causes approximately two thirds of all brain infarctions. We previously demonstrated that statins protect from cerebral ischemia by upregulation of endothelial type III nitric oxide synthase (eNOS), but the downstream mechanisms have not been determined. Therefore, we investigated whether antithrombotic effects contribute to stroke protection by statins. METHODS: 129/SV wild-type and eNOS knockout mice were treated with atorvastatin for 14 days (0.5, 1, and 10 mg/kg). eNOS mRNA from aortas and platelets was measured by reverse-transcriptase polymerase chain reaction. Platelet factor 4 (PF 4) and beta-thromboglobulin (beta-TG) in the plasma were quantified by ELISA. Transient cerebral ischemia was induced by filamentous occlusion of the middle cerebral artery followed by reperfusion. RESULTS: Stroke volume after 1-hour middle cerebral artery occlusion/23-hour reperfusion was significantly reduced by 38% in atorvastatin-treated animals (10 mg/kg) compared with controls. Serum cholesterol levels were not affected by the treatment. eNOS mRNA was significantly upregulated in a dose-dependent manner in aortas and in thrombocytes of statin-treated mice compared with controls. Moreover, indices of platelet activation in vivo, ie, plasma levels of PF 4 and beta-TG, were dose-dependently downregulated in the treatment group. Surprisingly, atorvastatin-treatment did not influence PF 4 and beta-TG levels in eNOS knockout mice. CONCLUSIONS: The synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor atorvastatin upregulates eNOS in thrombocytes, decreases platelet activation in vivo, and protects from cerebral ischemia in normocholesterolemic mice. Antithrombotic and stroke-protective effects of statins are mediated in part by eNOS upregulation. Our results suggest that statins may provide a novel prophylactic treatment strategy independent of serum cholesterol levels. PMID- 11022079 TI - Cell death after exposure to subarachnoid hemolysate correlates inversely with expression of CuZn-superoxide dismutase. AB - BACKGROUND AND PURPOSE: Subarachnoid hemolysate (SAH) has been associated with oxidative brain injury, cell death, and apoptosis. We hypothesized that over expression of CuZn-superoxide dismutase (CuZn-SOD) would protect against injury after SAH, whereas reduction of its expression would exacerbate injury. METHODS: Saline (n=16) or hemolysate (n=50) was injected into transgenic mice overexpressing CuZn-SOD (SOD1-Tg), CuZn-SOD heterozygous knockout mutants (SOD1+/ ), and wild-type littermates (Wt). Mice were killed at 24 hours. Stress gene induction was evaluated by immunocytochemistry and Western blotting for hemeoxygenase-1 and heat shock protein 70. Apoptosis was evaluated by 3'-OH nick end-labeling and DNA gel electrophoresis. Cell death was quantified through histological assessment after cresyl violet staining. RESULTS: Histological assessment demonstrated neocortical cell death in regions adjacent to the blood injection. Overall cell death was reduced 43% in SOD1-Tg mutants (n=6) compared with Wt littermates (n=6; P<0.02). In contrast, cell death was increased >40% in SOD1+/- mutants (n=6; P<0.05). Both hemeoxygenase-1 and heat shock protein 70 were induced after SAH. Apoptosis was also present after SAH, as evidenced by 3' OH end-labeling and DNA laddering. However, the degree of stress gene induction and apoptosis did not vary between Wt, SOD1-Tg, and SOD1+/- mice. CONCLUSIONS: The extent of CuZn-SOD expression in the cytosol correlates with cell death after exposure to SAH in a manner separate from apoptosis. Overexpression of CuZn-SOD may potentially be an avenue for therapeutic intervention. PMID- 11022080 TI - Selective effects of subarachnoid hemorrhage on cerebral vascular responses to 4 aminopyridine in rats. AB - BACKGROUND AND PURPOSE: We postulated that some abnormalities in cerebrovascular function after subarachnoid hemorrhage (SAH) may involve underlying alterations in K(+) channel function. Thus, using pharmacological inhibitors, we assessed the influence of SAH on function of 2 types of K(+) channel in regulation of basilar artery diameter in vivo and membrane potential (E(m)) in vitro. METHODS: Rats were injected with saline (control) or autologous blood (SAH) into the cisterna magna. Two days later, effects of vasoactive drugs on the basilar artery were examined with a cranial window preparation. Vascular responses to 4-aminopyridine (4-AP), 3-aminopyridine (3-AP), tetraethylammonium (TEA), serotonin, acetylcholine, and adenosine were compared in control and SAH rats. Additional studies using intracellular microelectrodes evaluated the effects of 4-AP and serotonin on E(m) of basilar arteries isolated from control and SAH rats. RESULTS: Baseline artery diameter was 236+/-5 micrometer in control rats and 220+/-7 micrometer in SAH rats (P:<0. 05). 4-AP, but not 3-AP, constricted the basilar artery in control rats, and responses to 4-AP were reduced in SAH rats. Constrictor responses to TEA or serotonin were unaffected by SAH. Vasodilator responses to acetylcholine were impaired in SAH rats, whereas responses to adenosine were not different. Resting E(m) was -81+/-3 mV in control arteries and -79+/-3 mV in SAH arteries. Both 4-AP and serotonin depolarized the basilar artery, but only 4-AP-induced depolarization was impaired in SAH arteries. CONCLUSIONS: These data suggest that 4-AP induces cerebral vasoconstriction in vivo through smooth muscle depolarization due to inhibition of voltage-dependent K(+) channels. Furthermore, function of these K(+) channels may be selectively reduced in the basilar artery after SAH and thus could contribute to cerebral vascular dysfunction. PMID- 11022081 TI - Experimental radiosurgery simulations using a theoretical model of cerebral arteriovenous malformations. AB - BACKGROUND AND PURPOSE: A novel biomathematical arteriovenous malformation (AVM) model based on electric network analysis was used to investigate theoretically the potential role of intranidal hemodynamic perturbations in elevating the risk of rupture after simulated brain AVM radiosurgery. METHODS: The effects of radiation on 28 interconnected plexiform and fistulous AVM nidus vessels were simulated by predefined random or stepwise occlusion. Electric circuit analysis revealed the changes in intranidal flow, pressure, and risk of rupture at intervals of 3 months during a 3-year latency period after simulated partial/complete irradiation of the nidus using doses <25 and >/=25 Gy. An expression for risk of rupture was derived on the basis of the functional distribution of the critical radii of component vessels. The theoretical effects of radiation were also tested on AVM nidus vessels with progressively increasing elastic modulus (E:) and wall thickness during the latency period, simulating their eventual fibrosis. RESULTS: In an AVM with E=5. 0x10(4) dyne/cm(2), 4 (14.3%) of a total 28 sets of AVM radiosurgery simulations revealed theoretical nidus rupture (risk of rupture >/=100%). Three of these were associated with partial nidus coverage and 1 with complete treatment. All ruptures occurred after random occlusion of nidus vessels in AVMs receiving low-dose radiosurgery. Intranidal hemodynamic perturbations were observed in all cases of AVM rupture; the occlusion of a fistulous component resulted in intranidal rerouting of flow and escalation of the intravascular pressure in adjacent plexiform components. Risk of rupture was found to correlate with nidus vessel wall strength: a low E: of 1.9x10(4) dyne/cm(2) resulted in a 92.8% incidence of AVM rupture, whereas a higher E: of 7.0x10(4) dyne/cm(2) resulted in only a 3.6% incidence of AVM rupture. A dramatic reduction in rupture incidence was observed when increasing fibrosis of the nidus was modeled during the latency period. CONCLUSIONS: It was found that the theoretical occurrence of AVM hemorrhage after radiosurgery was low, particularly when radiation-induced fibrosis of nidus vessels was considered. When rupture does occur, it would appear from a theoretical standpoint that the occlusion of intranidal fistulas or larger-caliber plexiform vessels could be a significant culprit in the generation of critical intranidal hemodynamic surges resulting in nidus rupture. The described AVM model should serve as a useful research tool for further theoretical investigations of cerebral AVM radiosurgery and its hemodynamic sequelae. PMID- 11022082 TI - Angiotensin II AT(1) blockade normalizes cerebrovascular autoregulation and reduces cerebral ischemia in spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: Angiotensin II, through stimulation of AT(1) receptors, not only controls blood pressure but also modulates cerebrovascular flow. We sought to determine whether selective AT(1) antagonists could be therapeutically advantageous in brain ischemia during chronic hypertension. METHODS: We pretreated spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto controls with the AT(1) antagonist candesartan (CV-11974), 0.5 mg/kg per day, for 3 to 14 days, via subcutaneously implanted osmotic minipumps. We analyzed cerebral blood flow by laser-Doppler flowmetry, cerebral stroke in SHR after occlusion of the middle cerebral artery with reperfusion, and brain AT(1) receptors by quantitative autoradiography. RESULTS: Candesartan treatment normalized blood pressure and the shift toward higher blood pressures at both the upper and lower limits of cerebrovascular autoregulation in SHR. Candesartan pretreatment of SHR for 14 days partially prevented the decrease in blood flow in the marginal zone of ischemia and significantly reduced the volume of total and cortical infarcts after either 1 or 2 hours of middle cerebral artery occlusion with reperfusion, relative to untreated SHR, respectively. This treatment also significantly reduced brain edema after 2 hours of middle cerebral artery occlusion with reperfusion. In SHR, candesartan markedly decreased AT(1) binding in areas inside (nucleus of the solitary tract) and outside (area postrema) the blood-brain barrier and in the middle cerebral artery. CONCLUSIONS: Pretreatment with an AT(1) antagonist protected hypertensive rats from brain ischemia by normalizing the cerebral blood flow response, probably through AT(1) receptor blockade in cerebral vessels and in brain areas controlling cerebrovascular flow during stroke. PMID- 11022083 TI - Role of phosphatidylinositol 3-kinase in acetylcholine-induced dilatation of rat basilar artery. AB - BACKGROUND AND PURPOSE: We tested the hypothesis that activation of phosphatidylinositol (PI) 3-kinase is involved in dilator responses of the basilar artery to acetylcholine in vivo. METHODS: Responses of the basilar artery were measured by the cranial window technique in anesthetized rats. To examine the role of PI 3-kinase in acetylcholine-induced calcium signaling, we measured intracellular free calcium concentration ([Ca(2+)](i)) of cultured rat basilar arterial endothelial cells using a fluorescent calcium indicator, indo 1. RESULTS: -Topical application of acetylcholine (10(-6), 10(-5.5), and 10(-5) mol/L) increased the diameter of the basilar artery by 8+/-1%, 14+/-2%, and 24+/ 3%, respectively. An inhibitor of PI 3-kinase, wortmannin (10(-8) mol/L), did not change the baseline diameter of the artery. In the presence of wortmannin, acetylcholine (10(-6), 10(-5.5), and 10(-5) mol/L) dilated the artery only by 3+/ 2%, 6+/-2%, and 12+/-2%, respectively. Thus, wortmannin attenuated acetylcholine induced dilatation of the basilar artery (P<0.05 versus control). Wortmannin had no effect on dilatation of the artery in response to a nitric oxide donor, sodium nitroprusside. LY294002, another inhibitor of PI 3-kinase, also inhibited dilator response of the basilar artery to acetylcholine. Acetylcholine produced an increase in [Ca(2+)](i) of the endothelial cells. Genistein, an inhibitor of tyrosine kinase, markedly attenuated acetylcholine-induced calcium influx to the cells; however, wortmannin had no effect on acetylcholine-induced calcium changes. CONCLUSIONS: These results suggest that acetylcholine-induced dilatation of the basilar artery is mediated, at least in part, by activation of PI 3-kinase in vivo. Acetylcholine-induced [Ca(2+)](i) changes of the endothelial cells may not be mediated by activation of the kinase. PI 3-kinase as well as [Ca(2+)](i) may play an important role in the acetylcholine-induced nitric oxide production of the basilar arterial endothelial cells. PMID- 11022084 TI - Inapparent hemodynamic insufficiency exacerbates ischemic damage in a rat microembolic stroke model. AB - BACKGROUND AND PURPOSE: Patients with severe carotid artery stenosis may have more severe ischemic damage after embolic stroke than patients without this abnormality. Unilateral proximal carotid occlusion (UCO) alone typically does not induce infarction in normotensive rats. The aim of this study was to investigate whether UCO increases infarct size after microembolic, experimental stroke. METHODS: Microembolic infarction was induced in 2 groups of Sprague-Dawley rats by injecting 2000 microspheres (50-micrometer diameter) intracranially from the external carotid artery. The common carotid artery (CCA) was either ligated just after the injection (CCA occlusion group, n=8) or left intact (CCA open group, n=8). In the control group (n=4), vehicle without microspheres was injected and the CCA was ligated. Twenty-four hours later, the brains were removed and infarct volumes measured. Perfusion-weighted imaging was used to evaluate the cerebral circulation before and after CCA occlusion with and without microsphere injection in a separate group of animals (n=16). RESULTS: All animals in the microemboli groups survived and had only a slight hemiparesis 24 hours after occlusion. No neurological deficits were observed in the control group. Infarct volumes were 145+/-57 mm(3) in the CCA occlusion group and 45+/-26 mm(3) in the CCA open group (P <0.01). There were no infarctions detected in the control group. Perfusion weighted imaging showed that cerebral blood flow decreased after the CCA occlusion in both experiments with and without the microsphere injection. CONCLUSIONS: UCO alone does not induce ischemic damage, but it worsens ischemic lesion size after multiple microemboli. This is probably due to the slight cerebral perfusion insufficiency caused by UCO. These results suggest that patients with cerebral hemodynamic insufficiency, such as those with severe carotid stenosis, may have increased ischemic damage after microembolic events. PMID- 11022085 TI - Extracellular pH, Ca(2+) influx, and response of vascular smooth muscle cells to 5-hydroxytryptamine. AB - BACKGROUND AND PURPOSE: Cerebral vascular smooth muscle cells (VSMCs) contract on extracellular pH (pH(o)) increases and relax on pH(o) decreases. These changes in tone are believed to result from changes in [Ca(2+)](i), although the responsible mechanisms are not fully understood. VSMCs also contract in response to 5 hydroxytryptamine (5-HT), which increases [Ca(2+)](i) via both Ca(2+) release and influx. We hypothesized that examining effects of pH(o) decreases on 5-HT-induced [Ca(2+)](i) changes would allow us to identify mechanisms whereby pH(o) influences tone. Accordingly, we compared [Ca(2+)](i) increases in cerebral VSMCs, evoked by 5-HT, with increases evoked by increased pH(o) and examined 5-HT dependent [Ca(2+)](i) increases at normal and decreased pH(o). METHODS: We monitored [Ca(2+)](i,), using the Ca(2+)-sensitive dye fura 2, in cultured rat cerebral VSMCs obtained by enzymatic digestion of middle cerebral arteries and their branches (passages 1 to 3) grown on glass coverslips and superfused with physiological saline. RESULTS: Increasing pH(o) from 7.3 to 7.8 increased [Ca(2+)](i), and these increases were prevented in Ca(2+)-free solutions. Decreasing pH(o) from 7.3 to 6.9 did not alter [Ca(2+)](i) unless [Ca(2+)](i) was first raised by treatment with 5-HT (10 micromol/L). 5-HT resulted in biphasic [Ca(2+)](i) increases characterized by transient peaks blocked by the Ca(2+) ATPase inhibitor thapsigargin (10 nmol/L) and prolonged plateaus blocked by the Ca(2+) channel blocker Ni(2+) (1 mmol/L). Acidification did not alter the transient peaks but significantly reduced 5-HT-induced Ca(2+) influx. CONCLUSIONS: We conclude that increasing pH(o) induces Ca(2+) influx in rat cerebral VSMCs and decreasing pH(o) inhibits 5-HT-stimulated Ca(2+) entry but not intracellular Ca(2+) release. PMID- 11022086 TI - Cerebral microembolism detected by transcranial Doppler during cardiac procedures. AB - BACKGROUND AND PURPOSE: Cerebral embolism with clinical sequelae may rarely complicate cardioversion and cardiac catheterization. Transcranial Doppler sonography has recently been introduced to monitor microemboli entering the middle cerebral artery in cardiac and carotid surgery. We therefore used this technique to evaluate the risk of asymptomatic embolism during common cardiac procedures. METHODS: Patients were monitored by transcranial Doppler while undergoing direct current cardioversion (n=15) and cardiac catheterization (n=17). RESULTS: Microemboli were detected in all patients having cardiac catheterization but in only 1 patient after cardioversion. CONCLUSIONS: Microembolism occurred frequently during cardiac catheterization and rarely during cardioversion. It is not yet known whether this has clinical relevance. PMID- 11022087 TI - Surgery in intracerebral hemorrhage. The uncertainty continues. AB - BACKGROUND AND PURPOSE: Primary intracerebral hemorrhage (ICH) accounts for 10% to 20% of stroke but carries the highest rates of mortality and morbidity of all stroke subtypes. Current treatment, however, is varied and haphazard. The most recent Cochrane systematic review refers to 4 prospective, randomized controlled trials. We present a further meta-analysis to include 3 new trials. In addition, we review the trials of Chen et al and McKissock et al and discuss aspects of their quality that, we believe, prevent their inclusion in modern day meta analysis. METHODS: Literature databases and articles were searched from 1966 to October 1999. Using the end points of death and dependency, the results of the 7 identified randomized trials were expressed as odds ratios. All available data were then analyzed with meta-analysis techniques. Analysis of relevant subsets of trials was also carried out. RESULTS: Meta-analysis of all 7 trials shows a trend toward a higher chance of death and dependency after surgery (OR 1.20; 95% CI 0.83 to 1.74). Meta-analysis was also carried out after exclusion of the Chen and McKissock trials for reasons discussed in the text. This meta-analysis suggests a benefit from surgery, with a reduction in the chances of death and dependency after surgical treatment by a factor of 0.63 (OR 0.63; 95% CI 0.35 to 1.14). CONCLUSIONS: When meta-analysis is restricted to modern-day, post-CT, well constructed, balanced trials, a trend for surgery to reduce the chances of death and dependency is found. Perhaps, then, in the modern era of CT, good neuroanesthesia, intensive care, and the operating microscope, surgery has a role in the treatment of supratentorial intracerebral hemorrhage. The results of a large, multicenter, randomized controlled trial are urgently needed, and the ongoing International Surgical Trial of Intracerebral Hemorrhage should fulfill this objective. PMID- 11022088 TI - Fatal hemorrhagic transformation of acute cerebral infarction after the use of abciximab. PMID- 11022089 TI - Chlamydia pneumoniae infection and PFO-associated ischemic stroke. PMID- 11022090 TI - There's no place like home...for some. PMID- 11022091 TI - Systematic comparison of the early outcome of angioplasty and endarterectomy for symptomatic carotid artery disease. PMID- 11022092 TI - The time concept in ischemic stroke: misleading. PMID- 11022094 TI - Abstracts of literature PMID- 11022093 TI - Association between physiological homeostasis and early recovery after stroke. PMID- 11022096 TI - En juin dernier PMID- 11022095 TI - Major Ongoing Stroke Trials. PMID- 11022097 TI - [Vitamin E and cardiovascular prevention]. AB - The antioxidant properties of vitamin E are well established. In humans, they appear very clearly from the nutritional supplementation trials. There is a strong correlation between supplied doses (>= 50 mg/j), vitamin E content of LDL and antioxidant protection of LDL. The consumption studies mostly suggest that the cardiovascular disease risk is diminished by the vitamin E supplementation, this being not true for vitamin E supplied by food strictly. In spite of the fact that there is a coherence between these results due in particular to the highly atherogenic role of oxidized low density lipoprotein, it is not allowed to claim that only the increased protection of LDL against oxidation is responsible for the diminished risk. The cell-regulating properties of vitamin E that have been more recently discovered have also to be taken into account as regards the functions of platelets, monocytes-macrophages, endothelial cells and vascular smooth muscle cells. The LDL-vitamin E capacity at decreasing the superoxide anion production (involved in turn in the oxidation process of LDL) could also play a role in preventing cardiovascular risk. The nutritional intervention studies undertaken in secondary prevention indicate a beneficial effect in terms of cardiovascular morbidity, either for low dose (50 mg), or for higher dose (>= 270 mg/d) intake, but without effect in terms of mortality. A recent study presumably supports a beneficial effect at the dose intake of 300 mg/d only in terms of cardiovascular mortality. Only one intervention has been carried out in condition of primary prevention, leading to the absence of effect at the dose employed (50 mg/d). The studies on the mechanisms of action make plausible the beneficial effects observed in analytical or experimental epidemiology. However, the experimental epidemiology does not provide indisputable evidence for the efficacy of the secondary prevention of cardiovascular risk by vitamin E supplementation. There is no intervention study using doses higher than 50 mg/d in primary prevention. There is a need for such studies in the not too distant future. A period of several years will be necessary before having new data possibly resulting in a consensus achievement. PMID- 11022098 TI - [Dendritic cells: orchestration of the immune response]. AB - Dendritic cells are critical initiators of immune responses. They not only activate pathogen-specific T and B lymphocytes, they also stimulate cells of the innate immune system. Furthermore, dentritic cells are involved in immunological tolerance induction through the elimination of autoreactive T lymphocytes. Over the last ten years, understanding of the development, biology and physiopathology of dentritic cells has progressed considerably which has lead to the use of dentritic cells in immunotherapy protocols to treat tumors, infections and auto immune diseases. PMID- 11022099 TI - [Antiphospholipid antibodies: clinical significance and biological diagnosis]. AB - The term "antiphospholipids" (aPLs) refers to an heterogeneous family of antibodies diagnosed either by clotting tests: the lupus anticoagulants or by Elisa: anticardiolipin (aCL) and anti-beta2-glycoprotein I (anti-beta2GP1) especially. aPLS recognize phospholipids, alone or bound to plasma protein cofactor(s), or the cofactors themselves. aPLs have long been described in autoimmune diseases such as SLE, but may also be found in other clinical settings including infections, malignancies and drug administration. Their persistent presence can be associated with venous and/or arterial thrombotic complications and/or recurrent miscarriage, thus defining the "antiphospholipid syndrome" (APS). The heterogeneity of aPLs makes a comprehensive approach to laboratory investigation essential. Detection of lupus anticoagulants relies on increased clotting times in phospholipid-dependent tests. Their 4 step diagnosis includes: 1) screening (by at least two different tests); 2) demonstration of an inhibitory activity; 3) evidence of its phospholipid dependence; 4) exclusion of an associated coagulopathy. Among the aPLs detected by Elisa, IgG aCL are the most frequently investigated. However, other antibodies may represent useful biological tools. Among them, anti-beta2GP1 are thought to be more closely associated with a history of thrombosis than aCL and testing for anti-beta2 GP1 should now be systematically included in the biological diagnosis of APS. The Elisa used for aCL and anti-beta2GP1 are not fully standardized, and a number of methodological parameters may account for the interlaboratory discrepancies often observed. The clinical importance of other antibodies such as antiphosphatidylethanolamine, antiprothrombin or antiannexin V is being evaluated. An appropriate laboratory investigation of APS should, in all cases, combine the use of clotting and immunological assays, and assess the persistence of autoantibodies over time. PMID- 11022100 TI - [Follow-up of the treatment by direct thrombin inhibitors: activated partial thromboplastin time or ecarin clotting time]. AB - The clinical use of the direct inhibitors of thrombin requires a reliable test to monitor the treatment and to predict the hemorragic risk. The activated partial thromboplastin time (APTT) is the most common test used to monitor treatment with unfractionated heparin. Thus APTT has been first chosen to follow patients treated with direct thrombin inhibitors, but studies have shown that it was probably not the most appropriate test. Indeed, APTT values were not well correlated with the dose administered and were dependent on the type of the thrombin inhibitor used and on the APTT reagent. The ecarin clotting time (ECT), which converts prothrombin into meizothrombin has been then tested and seemed to be a better test. In vitro studies have shown a good correlation between ECT and the different concentrations of thrombin inhibitors. Furthermore, the ECT in contrast to APTT is not sensitive to heparin or oral anticoagulant and interindividual variations are low with ECT. ECT which is a reliable test and is easy to perform seems to be a more appropriate test to monitor treatment with direct thrombin inhibitors but further studies are needed to validate its use in a clinical setting. PMID- 11022102 TI - [Studies on the genotoxic effects of crude liver oils from 3 species of Mediterranean sharks by means of in vitro micronucleus test using human lymphocytes]. AB - Lymphoid system tumours have been identified in two subjects who used to handle for several years mediterranean shark liver oil and squalen extracted from this oil. Moreover, scientific data, reported in 1959 by Kroning, show the induction of lymphoid tumours in C57 B1 mice after exposure of their skin to squalen. These observations rose the question of a possible mutagenic power of shark liver oil. In order to determine the genotoxicity of these oils, in vitro assays have been performed on crude hepatic oil of three species of mediterranean sharks: two benthic sharks, Centrophorus granulosus and Galeus melastomus, and one pelagic specie, Prionace glauca. Genotoxicity of oils have been assayed using a micronucleus test which can detected simultaneously clastogen and aneugen effects. The incubation of human cells with the hepatic crude oils of Centrophorus granulosus increases the rate of the binucleated micronucleated cell in a dose dependent manner. The mean micronucleated cell rate was 9.0%. +/- 1.1 in controls and increased up to 27,1%. +/- 4,0 for the highest concentrations of oil extracts. Similar results have been obtained with crude hepatic oils of Galeus melastomus and Prionace glauca. The results of this experimental study show that the crude liver oils of three species of sharks are genotoxic and confirm a high carcinogenic risk. PMID- 11022101 TI - [Search of biological markers of Alzheimer's disease]. AB - Peripheral markers for Alzheimer's disease are of interest to confirm the diagnosis, to perform epidemiological screening, to identify distinct groups of patients, to predict the outcome of the disease, to monitor its progression and its sensibility to treatment and to give help in performing studies on the relationship between brain and behaviour and on the pathophysiology of the Alzheimer's disease. The ideal biomarker for Alzheimer's disease should detect a fundamental feature of neuropathology and be validated in neuropathologically confirmed cases and be confirmed by at least two independent studies; should be as sensitive and specific than the clinical diagnosis (about 85% and 80%), reliable, reproducible, simple to perform, inexpensive and non invasive (studies on blood, urine, saliva, or buccal scrapings) or moderately invasive (skin, rectal biopsies, bone marrow samples, or cerebrospinal fluid). Such a marker has not yet been found. In this paper we present those markers which come closest to fulfilling criteria for a useful biomarker, keeping in mind that these criteria depends on what purpose it is used (screening, prediction, diagnosis, monitoring, pathophysiological studies.) and that the finding of a good marker depends on the understanding of the disease. PMID- 11022103 TI - [Nosocomial infections due to rotavirus and respiratory syncytial virus in pediatric wards: a 2-year study]. AB - Rotavirus and respiratory syncytial virus (RSV) infections represent up to 30% of the totality of nosocomial infections in paediatric wards. We studied the importance of these infections in the paediatric wards of the University Hospital Center of Poitiers, France, from October 1996 to September 1998. We defined as nosocomial an infection acquired after 3 days of hospitalization for rotavirus and after 7 days for RSV. The 274 cases of children presenting rotavirus gastroenteritis or RSV infection within this period were studied. Rotavirus was detected in stools by using an agglutination test and RSV was diagnosed in nasopharyngeal aspirations by direct examination with an immunofluorescence assay (IFA), cell culture and serotyping with IFA. We noted 50 rotavirus and 224 RSV infections, with a first epidemic of RSV subgroup B (49.5%) and a second epidemic of subgroup A (44.9%). 19 (38%) were rotavirus nosocomial infections and 5 (2.2%) were RSV nosocomial infections. The majority of the nosocomial infections occurred before the age of one year and particularly before the age of 6 months (42.2% for rotavirus, 60% for RSV). In comparison to community-acquired infections, children with rotavirus nosocomial infections were younger (9 months versus 12.5 months) which was the opposite for RSV nosocomial infections (10.8 months versus 6.5 months). The sex-ratio of children with community-acquired infections was 2.1 that was not reported in nosocomial infections. The length of stay in hospital was always longer in nosocomial infections (11.7 days versus 3.6 days for rotavirus; 38.8 days versus 4.8 days for RSV). Diarrhea (p = 0.007) and vomiting (p = 0.013) for enteric infections and wheezing (p = 0.02) for respiratory infections were more often observed in community-acquired infections. This study emphasizes the frequency and the consequences of rotavirus and RSV nosocomial infections in paediatric wards and the importance of the hygienic rules to prevent these infections. PMID- 11022104 TI - [Pseudohypertriglyceridemia due to hyperglycerolaemia]. PMID- 11022105 TI - [Isolation of Pneumococcus in the bile]. PMID- 11022106 TI - [Evaluation of the detection of procalcitonin by an immuno-chromatography test: Brahms PCT-Q]. PMID- 11022107 TI - [Biological markers of renal dysfunction in essential arterial hypertension in African subjects]. PMID- 11022108 TI - [Value of a new method of LDL cholesterol assay in case of moderate hypertriglyceridemia]. PMID- 11022109 TI - [Case of massive pulmonary hemorrhage due to Leptospira icterohaemorrhagiae with a fatal outcome]. PMID- 11022110 TI - [An original case of asymptomatic erythrocytosis]. PMID- 11022111 TI - [Control of the quality of drug doses in the framework of therapeutic surveillance: Asqualab program 1997-1999]. PMID- 11022112 TI - [Hemolysis and blood proteins]. PMID- 11022113 TI - ["Hemolyzed" blood sample]. PMID- 11022114 TI - [The point of SI Units]. PMID- 11022116 TI - Assemblee generale de la Societe francaise de biologie clinique PMID- 11022115 TI - [Comments on analysis reports by the biologist: Why? How? For whom? Plea for a debate]. PMID- 11022117 TI - Cell contact-mediated signaling of monocytes by stimulated T cells: a major pathway for cytokine induction. AB - T lymphocytes are currently thought to play a pivotal part in the pathogenesis of chronic inflammatory diseases. However, the mechanism(s) by which they exert their pathogenic effect remain(s) elusive. Contact-mediated signaling of monocytes by stimulated T cells is a potent pro-inflammatory mechanism that triggers massive up-regulation of the pro-inflammatory cytokines interleukin-1 (IL-1) and tumor necrosis factor-a (TNF-alpha) that play an important part in chronic destructive diseases such as rheumatoid arthritis and multiple sclerosis. To date cell-cell contact is the only endogenous mechanism to be described that displays such an activity in monocyte-macrophages which are classically stimulated in vitro by bacterial products such as LPS or non-specific stimuli such as phorbol esters or poorly activated by soluble cytokines such as IFN gamma. Since direct cellular contact occurs at the inflammatory site, we hypothesized that this mechanism is relevant to the pathogenesis of chronic inflammatory disorders. This review aims at summarizing the state of the art and importance of contact-mediated monocyte activation by stimulated T lymphocytes. PMID- 11022118 TI - Invertebrate and fish cytokines. AB - Cytokine-like molecules are well described in invertebrates, although most recent studies have revealed that there is analogy, rather than homology, between invertebrate and vertebrate cytokine-like activities. Cytokines certainly appeared early in the evolution of vertebrates, dating back some 400 millions years. Here, evidence will be reviewed and updated of the presence of these molecules in jawed fish and in particular, in bony fish, which represent the oldest group displaying true functionality of immune system as known in modern vertebrates. Many studies during the last ten years have confirmed the presence of functional homologues of mammalian cytokines in fish. In this review, particular attention will be focussed on IL-1beta, a very ancient defence cytokine recently sequenced in two species, rainbow trout (Oncorhynchus mykiss) and carp (Cyprinus carpio). Original data on the partial peptide sequence of IL 1beta in the mediterranean sea bass Dicentrarchus labrax are also presented. PMID- 11022119 TI - Three novel mammalian toll-like receptors: gene structure, expression, and evolution. AB - We describe three novel genes, encoding members of the Toll-like receptor (Tlr) family (TLR7, TLR8, and TLR9). These Tlr family members, unlike others reported to date, were identified within a genomic database. TLR7 and TLR8 each have three exons, two of which have coding function, and lie in close proximity to one another at Xp22, alongside a pseudogene. The remaining gene (TLR9) resides at 3p21.3 (in linkage with the MyD88 gene), and is expressed in at least two splice forms, one of which is monoexonic and one of which is biexonic, the latter encoding a protein with 57 additional amino acids at the N-terminus. The novel Tlrs comprise a cluster as nearest phylogenetic neighbors. Combining all sequence data related to Toll-like receptors, we have drawn several inferences concerning the phylogeny of vertebrate and invertebrate Tlrs. According to our best estimates, mammalian TLRs 1 and 6 diverged from a common mammalian ancestral gene 95 million years ago. TLR4, which encodes the endotoxin sensor in present-day mammals, emerged as a distinct entity 180 million years ago. TLRs 3 and 5 diverged from a common ancestral gene approximately 150 million years ago, as did Tlr7 and Tlr8. Very likely, fewer Tlrs existed during early vertebrate evolution: at most three or four were transmitted with the primordial vertebrate line. Phylogenetic data that we have adduced in the course of this work also suggest the existence of a Drosophila equivalent of MyD88, and indicate that the plasma membrane protein SIGIRR is close functional relative of MyD88 in mammals. Finally, a single present-day representative of the Toll-like proteins in Drosophila has striking cytoplasmic domain homology to mammalian Tlrs within the cluster that embraces TLRs 1, 2, 4, and 6. This would suggest that an ancestral (pre-vertebrate) Tlr may have adopted a pro-inflammatory function 500 million years ago. PMID- 11022120 TI - Cloning and characterization of a sub-family of human toll-like receptors: hTLR7, hTLR8 and hTLR9. AB - Members of the Toll-like receptor family are essential components of the innate immune system. Herein we report the molecular cloning and characterization of three novel human Toll-like receptors (hTLRs) designated hTLR7, hTLR8, and hTLR9. Human TLR7-9, like the previously described members hTLR1-6 contain an ectodomain with multiple leucine-rich repeats (LRRs) and a cytoplasmic domain homologous to that of the human interleukin-1 (IL-1) receptor. When compared with hTLR1-6, the hTLR7-9 has a higher molecular weight largely as a result of a longer ectodomain. Phylogenetic analysis shows that hTLR7-9 belong to a new sub-family of the hTLRs. Analysis of mRNA expression at the tissue levels shows differential expression patterns; hTLR7 is predominantly expressed in lung, placenta and spleen, hTLR8 is more abundant in lung, peripheral blood leukocytes, and hTLR9 is preferentially expressed in immune cell rich tissues, such as spleen, lymph node, bone marrow and peripheral blood leukocytes. The hTLR7 and hTLR8 genes are located on the sex chromosome X, hTLR9 gene is located on chromosome 3. Expression of constitutively active hTLR7-9 stimulates an NF-kappaB signaling pathway indirectly supporting the contention that these receptors are involved in cellular responses to stimuli, which activate innate immunity. PMID- 11022121 TI - A short course of oral aspirin increases IL-18-induced interferon-gamma production in whole blood cultures. AB - The effect of aspirin on whole blood cytokine production was studied in six healthy volunteers. Four days after cessation of a 3-day regimen of 650 mg of oral aspirin, there was a 70% increase in interferon-gamma (IFN-gamma) production, stimulated by a combination of interleukin-18 (IL-18) plus lipopolysaccharide (p < 0.05). At this time, there was a 4-fold increase in the production of tumor necrosis factor-alpha (TNF-alpha) compared to pre-aspirin levels (p < 0.03). TNF-alpha and IFN-gamma production returned to pre-aspirin levels one month after the discontinuation of aspirin. Short-term aspirin treatment induces a significant increase in the production of these cytokines, probably through inhibition of prostaglandins. These data suggest a novel pathway through which long aspirin use reduces the risk of colon cancer, and may explain the effects of aspirin in inflammatory bowel disease. PMID- 11022122 TI - Human keratinocytes are major producers of IL-18: predominant expression of the unprocessed form. AB - The cytokine network in the skin is a tightly regulated system in which IL-1 isoforms, as well as their receptors and antagonists have a central role. The recently discovered IL-1 isoform IL-18 (also known as interferon gamma-inducing factor (IGIF) or IL-1gamma), promotes IFN-gamma expression by T cells in concert with IL-12. Because IFN-gamma plays an important role in many inflammatory skin diseases by facilitating the development of Th1 cells, it is important to elucidate the role of mediators which regulate the production of this cytokine. We demonstrate that human keratinocytes constitutively express IL-18 at the mRNA as well as at the protein level. The protein was mainly expressed intracellularly in the 24 kD unprocessed pro-form, but was also secreted. Histochemistry revealed a diffuse staining of IL-18 in the epidermis of normal skin, which is in line with our in vitro data. Furthermore, we show that the level of IL-18 expressed in freshly isolated normal human epidermal cells, whether or not containing HLA-DR+ cells, significantly exceeded the expression levels of other cell types such as monocytes and bronchial epithelial cells. Finally, our results show that stimulation of the keratinocyte cell line HaCaT with PMA LPS or IL-1beta, does not significantly affect intracellular or released (pro) IL-18 levels. These experiments show for the first time that human keratinocytes relative to monocytes, PBMC or leukocytes produce a considerably larger amount of pro-IL-18, which is also readily released. High constitutive levels of IL-18 may contribute to the skewing towards a Th1-like environment, which is apparent in many human inflammatory skin diseases. PMID- 11022123 TI - Relation of pro- and anti-inflammatory cytokines and the production of nitric oxide in patients receiving high-dose immunotherapy with interleukin-2. AB - Immunotherapy with intravenous recombinant human interleukin-2 (rh IL-2) may be accompanied by hypotension and the emergence of capillary leak syndrome. Nitric oxide (NO) is supposed to be responsible for both side effects. The aim of the current investigation was to elucidate the relationship between pro- and anti inflammatory cytokines and the production of NO in eight tumor patients receiving intravenous rh IL-2 continuously over a time period of 120 hours. Markers of systemic inflammation, as well as nitrate plasma levels, were consecutively determined. Significant changes in the levels of pro-inflammatory cytokines IL-6 and IL-8 were observed (p < 0.05). In contrast to the anti-inflammatory cytokine IL-10, which did not increase significantly, the serum concentrations of the soluble tumor necrosis factor receptors (sTNFr) I and II rose continuously and significantly during the observation period (p < 0.05). In parallel, a significant rise in nitrate plasma levels was observed (p < 0.05). Moreover, there were highly significant correlations between nitrate and IL-6 serum levels (p < 0.05), nitrate and sTNFr-I (p < 0.05), nitrate and sTNFr-II (p < 0.05), and between IL-6 and IL-10 (p < 0.05), respectively. We conclude that immunotherapy with IL-2 promotes a pro-inflammatory state, parallelled by an increased production of nitric oxide. Although anti-inflammatory responses accompany this process, they are not able to diminish the production of nitric oxide. PMID- 11022124 TI - Lower interleukin-2 and higher serum tumor necrosis factor-a levels are associated with perimenstrual, recurrent, facial Herpes simplex infection in young women. AB - The aim of this study was to look at a possible relationship between the recurrent perimenstrual dermatosis - facial Herpes simplex infection and the serum concentrations of interleukin-2 (IL-2) and tumor necrosis factor alpha (TNF alpha). Twenty-one volunteers (19-26 year olds) were examined at five points of the menstrual cycle. Ten volunteers were characterised by recurrent Herpes simplex infection lasting either from the 18th or the 25th day of the menstrual cycle until a few days after menstruation. Eleven young women without symptoms formed the control group. Both groups were similar as regards blood levels of 17beta-estradiol and progesterone. The group with the frequent infectious symptoms was characterised, however, by lower concentrations of IL-2 throughout the whole menstrual cycle, as compared to those without the symptoms. Levels of IL-2 in this group additionally dropped significantly on the 18th and on 25th day of the cycle. Moreover, the group with symptoms was characterised by higher level of TNF-alpha on the 18th day. These changes were found during the menstrual cycle of the women with recurrent herpes infection who however, at the time of the examination were free of the clinical symptoms. There was a similar tendency in both groups towards an increase in the levels of TNF-a around menstruation. Measurement of the other serum pro-inflammatory marker - IL-6 showed higher levels of this cytokine during the menstrual cycle in the group with the clinical symptoms. The results indicate that a decrease of IL-2 together with an increase of TNF-alpha and IL-6 in the serum seem to be related to recurrent perimenstrual Herpes simplex infection. PMID- 11022125 TI - Activated protein C inhibits tumor necrosis factor and macrophage migration inhibitory factor production in monocytes. AB - The precise regulatory mechanisms of amplification and downregulation of the pro- and anti-inflammatory cytokines in the inflammatory response have not been fully delineated. Although activated protein C (APC) and its precursor protein C (PC) have recently been reported to be promising therapeutic agents in the management of meningococcal sepsis, direct evidence for the anti-inflammatory effect remains scarce. We report that APC inhibits in vitro the release of tumor necrosis factor (TNF) and macrophage migration inhibitory factor (MIF), two known cytokine mediators of bacterial septic shock, from lipopolysaccharide (LPS)-stimulated human monocytes. The THP-1 monocytic cell line, when stimulated with LPS and concomitant APC, exhibited a marked reduction in the release of TNF and MIF protein in a concentration-dependent manner compared to cells stimulated with LPS alone. This effect was observed only when incubations were performed in serum free media, but not in the presence of 1-10% serum. Serum-mediated inhibition could only be overcome by increasing APC concentrations to far beyond physiological levels, suggesting the presence of endogenous serum-derived APC inhibitors. Inhibition of MIF release by APC was found to be independent of TNF, as stimulation of MIF release by LPS was unaltered in the presence of anti-TNF antibodies. Our data confirm that the suggested anti-inflammatory properties of APC are due to direct inhibition of the release of the pro-inflammatory monokine TNF, and imply that the anti-inflammatory action of APC is also mediated via inhibition of MIF release. PMID- 11022126 TI - Dexamethasone and cyclosporin A do not inhibit interleukin-15 expression in the human lung carcinoma cell line A549. AB - A549 cells constitutively expressed IL-15 mRNA which could be upregulated by stimulation with TNF-alpha- or IL-1beta. Constitutive and induced levels of IL-15 mRNA were not decreased in the presence of 10- 6 M dexamethasone. Control experiments revealed that 10- 6 M dexamethasone inhibited the TNF-alpha- or IL 1beta-mediated increase of IL-8 mRNA in A549 cells, which showed that the glucocorticoid was functional. A549 cells did not secrete relevant amounts of IL 15 protein. The constitutive expression and the TNF-alpha- or IL-1beta-mediated upregulation of intracellular IL-15 protein was not inhibited by dexamethasone, in contrast, the release of IL-8 protein was inhibited. Also, cyclosporin A at 250 ng/ml did not inhibit the TNF-alpha-induced upregulation of IL-15 mRNA and intracellular IL-15 protein. The data suggest that the synthesis of IL-15 mRNA and protein is not influenced by immunosuppressive glucocorticoids or by cyclosporin A. PMID- 11022127 TI - Sublethal hemorrhagic shock reduces tumor necrosis factor-alpha-producing capacity in different cell compartments. AB - Hemorrhagic shock results in a severe impairment of the immune response. Immunological alterations after hemorrhagic shock thus appear to be responsible for reduced resistance to infectious agents commonly observed after shock and severe injury. In the present study we examined the TNF-alpha-producing capacity of immune cells derived from different organs after sublethal shock in rats. Hemorrhagic shock was established by pressure controlled bleeding to a mean arterial pressure of 35 mm Hg for 35-40 min and consecutive resuscitation in male Sprague-Dawley rats. Twenty four hours after shock, TNF-a production in response to lipopolysaccharide (LPS, Salmonella friedenau) stimulation was measured in isolated spleen, bone marrow and blood cells. TNF-a production could be induced by stimulation with 1 ng/ml, in blood, spleen and bone marrow cells collected from sham-operated animals. A maximal stimulation was observed in all cell types after stimulation with 10 ng/ml LPS and could not be further increased with LPS doses of 100 ng/ml. Hemorrhagic shock of 35 mm Hg for 35-40 min, with consecutive resuscitation did not result in mortality, in contrast to a 4 hours lasting hemorrhagic shock resulting in 80% mortality. Blood, spleen or bone marrow cells, harvested from animals 24 hours after sublethal hemorrhagic shock, showed a significantly reduced TNF-alpha production in all cell populations after LPS stimulation. Serum collected from animals 2 hours after sublethal hemorrhagic shock contained an activity not present either before or 24 hours after shock, that downregulated LPS-induced TNF-alpha production in rat whole blood cultures and the murine macrophage cell line J774. The inhibitory activity present in serum, 2 hours after shock is not IL-10 since this mediator was not detectable in any serum sample. However, in the serum samples with TNF-alpha-inhibitory activity, elevated levels of PGE2 metabolites were found, which suggests the involvement of prostaglandins in trauma-induced immunosuppression. Altered TNF-a expression might be partially explained by an inhibitory activity in the serum already present 2 hours after shock. Since adequate, but not overwhelming TNF alpha production is essential for host response, the altered cytokine formation might explain local and systemic susceptibility to infections after hemorrhagic shock. PMID- 11022128 TI - Interleukin-1 signaling in mouse astrocytes involves Akt: a study with interleukin-4 and IL-10. AB - Although astrocytes are well known to respond to the pro-inflammatory cytokine, interleukin-1 (IL-1), the receptor and post-receptor mechanisms that mediate IL-1 effects in this cell type are complex and need further investigation. Using electrophoretic mobility shift assay (EMSA), we show that IL-1beta-induced NFkappaB activation in primary culture of mouse astrocytes is mediated by the interaction of this cytokine with the IL-1 type I receptor/IL-1 receptor accessory protein complex, as demonstrated by the ability of blocking monoclonal antibodies against these receptors to attenuate NFkappaB activation. In addition to NFkappaB activation, IL-1beta is also able to phosphorylate Akt, as demonstrated by Western blot. The observation that addition of wortmanin, that specifically blocks Akt phosphorylation, also attenuates NFkappaB activation can be interpreted that Akt phosphorylation interacts with IL-1 signaling pathways. Furthermore, anti-inflammatory cytokines such as IL-4 and IL-10 that block IL-1b induced NFkappaB activation also attenuate IL-1beta-induced Akt phosphorylation, despite the fact that IL-4 and IL-10 in isolation induced Akt phosphorylation. All these findings point to an interaction between Akt and NFkappaB-dependent IL 1 signaling in the primary culture of astrocytes. PMID- 11022129 TI - Prolactin activates interferon regulatory factor-1 expression in normal lympho hemopoietic cells. AB - It has been proposed that prolactin (PRL) is a lympho-hemopoietic growth and differentiation factor. We show here by Western blotting that PRL-receptors (PRL R) are expressed in normal rat bone marrow and spleen cells. We also show that PRL stimulates the phosphorylation of the PRL-R-associated Janus tyrosine kinase (JAK)-2 in rat bone marrow and spleen cells. This leads to the activation and subsequent binding of signal transducer and activator of transcription (Stat) 5b to an interferon regulatory factor-1 (IRF-1) gamma activation sequence (GAS) as visualized by electromobility shift assay. As shown after reverse transcription of mRNA by polymerase chain reaction, PRL, at physiological concentrations (0.01 microg/ml), stimulates the expression of the IRF-1 gene in these normal cells. PRL could thus affect several aspects of the immune response. PMID- 11022130 TI - Relationship between circulating interleukin-10 (IL-10) with interleukin-6 (IL-6) type cytokines (IL-6, interleukin-11 (IL-11), oncostatin M (OSM)) and soluble interleukin-6 (IL-6) receptor (sIL-6R) in patients with multiple myeloma. AB - We investigated the serum concentration of the interleukin-10 (IL-10), along with cytokines of interleukin-6 (IL-6) family (IL-6, IL-11 and oncostatin M - OSM), as well as soluble receptor for IL-6 (sIL-6R), in 121 patients with multiple myeloma (MM) and 28 healthy subjects. We studied the interactions between IL-10 and other cytokines, and the receptor. The correlation between IL-10 and some clinical and laboratory parameters associated with the disease activity were also analysed. The IL-10 was detectable in all patients with multiple myeloma and in all controls. The IL-10 concentration was significantly increased in myeloma patients compared with healthy persons (mean - 7.09 and 2.1 pg/ml, respectively) (p = 0.008). The level of IL-10 correlated positively with the advanced stage of disease estimated according to the Salmon and Durie classification (I versus III stage - p = 0.03). Higher values of IL-10 were found in patients with the light chain disease, hypercalcaemia, and correlated with the elevated concentrations of C-reactive protein (CRP). IL-6 was detected in 117 of the 121 patients and in all controls. The concentration of IL-6 was statistically increased in MM patients compared with control group (mean - 16.06 and 4.49 pg/ml, respectively) (p = 0.01). We found a positive correlation between IL-10 and IL-6 serum levels in MM patients. The relationship, expressed as Spearman's rank sum coefficient (rho = 0.249, p = 0.006) was significant. IL-11 was detected in 26 of the 121 MM patients and in 3 of the 28 healthy subjects at the mean concentration of 1.2 and 0.6 pg/ml respectively (p > 0.05). OSM was at detectable levels in 51 of the 121 patients and in only 4 of the 28 controls (mean - 3.84 and 0.1 pg/ml, p = 0. 002). The correlation between IL-10 and IL-11 levels in MM patients was not significant, but there was a strong statistical correlation between IL-10 and OSM concentrations (rho= 0.327, p = 0.0002). The serum concentration of sIL-6R was measurable in all patients and all controls (mean - 66.00 and 39.57 ng/ml respectively), but the difference between these groups was not significant. We found significant, positive correlation between the levels of IL-10 and sIL-6R (rho= 0.233, p = 0.01). In conclusion, we state that the serum concentrations of IL-10, IL-6, OSM and sIL-6R in MM patients may be a useful markers for the evaluation of the disease activity. PMID- 11022131 TI - Limited effects of placental and pituitary growth hormone on cytokine expression in vitro. AB - The hypothesis that growth hormone (GH) can affect immune responses in man has been evaluated by monitoring cytokine expression in cultures from peripheral blood mononuclear cells, by enzyme-linked immunosorbent assay (ELISA) and ribonuclease protection assay, and in tonsillar cells by ELISA. In addition to pituitary GH (GH-N), the placental form (GH-V), differing from pituitary GH by 13 amino acids has also been tested. Only few effects reached statistical significance and were in no case greater than 15%. Pituitary GH slightly reduced IL-5 production and stimulated IFN-gamma production. The latter effect was also observed with prolactin and could thus be induced through the prolactin receptor. It is proposed that GH has no strong effects on the parameters investigated, possibly as a result of redundancy in the cytokine network. Alternatively, effects on leukocytes are mediated by other tissues such as the liver or are clear only in response to stronger challenges. PMID- 11022132 TI - Phenotypes and cytokine profiles of enriched blood dendritic cells in healthy individuals. AB - Dendritic cells (DC) are highly specialized for initiating adaptive immune responses and are capable of producing a wide variety of cytokines. However, cytokine profiles of the DC naturally present in human blood have received relatively little attention. The objective of this study was to investigate expression of surface markers and cytokines by blood DC not subjected to prolonged culture and/or polyclonal activation, to identify surface phenotypes of cytokine-expressing DC and to evaluate sex and age differences in cytokine profiles of DC. For this purpose, DC were enriched from blood of healthy donors by the use of the adherence method, and expression of surface molecules and intracellular IFN-g, IL-10, IL-12 and IL-15 was studied by flow cytometry. Enriched blood DC expressed higher levels of IFN-g, IL-12 and IL-15, compared to whole mononuclear cells (MNC) incubated for the same time. Expression of IFN-g and IL-12 was confined to the mature CD83+CD11c+ DC subset. Enriched DC from females' blood displayed higher levels of CD80, IL-10 and IL-15. Taken together, enriched blood DC spontaneously express larger amounts of IFN-g, IL-12 and IL-15 than MNC. Sex differences in expression of CD80, IL-10 and IL-15 may have a modulatory influence on immune responses in males and females. PMID- 11022133 TI - Neurosteroid levels are increased in vivo after LPS treatment and negatively regulate LPS-induced TNF production. AB - Neuroactive steroids such as dehydroepiandrosterone sulfate and pregnenolone inhibit lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) production. Corticosteroids not only inhibit TNF production but their levels are increased in vivo after endotoxin injection, thus representing a feedback system that limits TNF production. We wondered whether the same could be true for neuroactive steroids. Thus, the possibility that neuroactive steroids might be increased concomitantly to TNF induction in vivo in mice treated with LPS was investigated. Increased plasma and hippocampal levels of allopregnanolone (but not of dehydroepiandrosterone or pregnenolone) were found 90 min after LPS injection. Allopregnanolone and progesterone (IC50 10- 7 and 10- 9 M, respectively) also inhibited TNF production by mouse peritoneal macrophages in vitro at concentrations in the range of those detected in vivo. These findings suggest that neuroactive steroids may act as endogenous inhibitors of cerebral and systemic TNF production. PMID- 11022134 TI - SDF-1-induced activation of ERK enhances HIV-1 expression. AB - Chemokine receptors are not only able to bind chemokines but, together with CD4, they serve as an entry door for the human immunodeficiency virus type 1 (HIV-1). The signalling capacity of chemokine receptors, which is of fundamental importance for chemokine-induced chemotaxis, is not used by HIV-1 to enter a target cell, nor by chemokines or chemokine-derived ligands to inhibit viral entry. In addition, an ill-defined signal triggered by chemokines can, under some circumstances, lead to an increase in HIV-1 expression. We show here that, in infected cells, exposure to SDF-1 leads to an increased expression of a X4 strain of HIV-1. A similar increase can be induced by an N-terminal peptide of SDF-1 which had previously been shown to elicit an intracellular calcium response and to inhibit the entry of X4 strains of HIV-1. We demonstrate the involvement of extracellular signal-regulated kinases (ERK) in this phenomenon. SDF-1 activates ERK-1 and ERK-2 in Jurkat cells. In HeLa cells, ERK-2 only is activated by SDF-1 or by a SDF-derived peptide. This ERK activation can be blocked by pertussis toxin and by the MEK inhibitor U0126. Most importantly, SDF-1-dependent HIV-1 expression is abolished by pretreating the cells with pertussis toxin or with U0126. The consequences of this SDF-1-induced, ERK-dependent modulation of HIV-1 expression in infected cells may have a clinical relevance for eradicating latent viruses. PMID- 11022136 TI - 3rd Symposium on Cytokines and Apoptosis in the Cardiovascular System. Halle, Germany, 27-28 January 2000. Abstracts. PMID- 11022135 TI - Cytokine directed targeting in chronic arthritis. PMID- 11022137 TI - Reflex epilepsy and reflex seizures of the visual system: a clinical review. AB - Reflex epilepsy of the visual system is charecterised by seizures precipitated by visual stimuli. EEG responses to intermittent photic stimulation depend on the age and sex of the subject and on how stimulation is performed: abnormalities are commonest in children and adolescents, especially girls. Only generalised paroxysmal epileptiform discharges are clearly linked to epilepsy. Abnormal responses may occur in asymptomatic subjects, especially children. Photosensitivity has an important genetic component. Some patients are sensitive to patterns, suggesting an occipital trigger for these events. Myoclonus and generalised convulsive and nonconvulsive seizures may be triggered by visual stimuli. Partial seizures occur less often and can be confused with migraine. Although usually idiopathic, photosensitive epilepsy may occur in degenerative diseases and some patients with photosensitive partial seizures have brain lesions. Sunlight and video screens, including television, video games, and computer displays, are the commonest environmental triggers of photosensitive seizures. Outbreaks of triggered seizures have occurred when certain flashing or patterned images have been broadcast. There are regulations to prevent this in some countries only. Pure photosensitive epilepsy has a good prognosis. There is a role for treatment with and without antiepileptic drugs, but photosensitivity usually does not disappear spontaneously, and then typically in the third decade. PMID- 11022138 TI - Cortical silent period following transcranial magnetic stimulation in epileptic patients. AB - Cortical silent period (SP) following transcranial magnetic stimulation is mainly due to cortical inhibitory mechanisms. SP may have a value for detecting inhibitory mechanisms in epileptic patients with or without treatment. The aim of this study was to evaluate the effect of both the epilepsy and the antiepileptic medication on these inhibitory mechanisms. The subgroups studied consisted of (a) normal subjects, (b) unmedicated epileptic patients, (c) epileptic patients with uncontrolled seizures under medication, (d) epileptic patients with controlled seizures under medication. SP following transcranial magnetic stimulation was measured in all subjects. The SP values from shortest to the longest were in the following order: 1) normal subjects; 2) epileptic patients with controlled seizures under medication; 3) unmedicated epileptic patients; 4) epileptic patients with uncontrolled seizures under medication. Our findings probably indicate the enhanced interictal inhibitory mechanisms in epilepsy which is resistant to antiepileptic medication. PMID- 11022139 TI - Mesial temporal lobe epilepsy versus amygdalar epilepsy: late seizure recurrence after initially successful amygdalotomy and regained seizure control following hippocampectomy. AB - We summarise the concept of mesial temporal lobe epilepsy and the pros and cons in order to define amygdala epilepsy. We present a patient with stereotactically proven right amygdalar seizure onset, associated with fear and vegetative autonomic signs and symptoms as the most prominent clinical ictal features. Following a right stereotactic amygdalotomy, the patient experienced an 11-year seizure-free period. Similar, but not identical, semeiology of complex partial seizures then recurred. A right-sided selective hippocampectomy and excision of the previously lesioned amygdala was performed. Except for 2 complex partial seizures associated with withdrawal of antiepileptic drugs, the patient remained seizure-free 9.5 years. This case underscores the important role of the amygdala in generating the semiology, and raises several questions concerning the existence of "amygdalar epilepsy". The 11-year seizure-free period following the stereotactic destruction of the amygdala is a strong argument for this notion. The late seizure recurrence requiring a second operation might, however, be seen as an argument for the important role of the hippocampal formation in the syndrome of mesial temporal lobe epilepsy even when the amygdala has been identified as the seizure onset zone. The role of stereotactic amygdalotomy is briefly reviewed. PMID- 11022140 TI - Clinical findings, imaging characteristics and outcome in catastrophic post encephalitic epilepsy. AB - OBJECTIVES: The aim of this study is to characterize the clinical features and prognostic factors for intractable, post-encephalitic epilepsy. METHODS: We studied retrospectively 42 patients (26 men) evaluated between 1982 and 1999. MRI, neuropsychological findings, interictal and ictal scalp EEG were reviewed for all patients. Fifteen patients had additional stereo EEG (SEEG) studies. RESULTS: The mean age at encephalitis was 17 years (SD = 15.5); etiology was identified in 18 patients. During the acute illness, 79% had status epilepticus (SE) or recurrent seizures and 76% were in coma; mean Glasgow outcome scale (GOS) was 3.6 (SD = 0.8). The mean latency to seizure onset was 0.8 years (SD = 1.9). The majority (72%) presented with complex partial seizures with or without secondary generalization. According to interictal epileptiform findings and the predominant seizure onset pattern as found on scalp EEG, patients were unilateral temporal (UTLE) = 8, bilateral temporal (BTLE) = 12, and extratemporal/multifocal or generalized (ETMFE) = 22 patients. MRI atrophy and/or signal changes were unilateral temporal in 7 (18%), bilateral temporal in 5 (13%), multilobar/diffuse in 20 (51%), and absent in 7 (18%). ANOVA revealed significant differences in mean GOS between UTLE versus BTLE and ETMFE (4.7 versus 3.2 versus 3.6; p < 0.0001), but not in age at encephalitis. Latency to the first unprovoked seizure was shorter in patients with ETMFE compared to UTLE and BTLE (p < 0.01). Surgery was performed in 24 patients. The best outcome was obtained in UTLE (7/8 class I and II). In the others, outcome was poor in the majority (13/16 class III and IV). CONCLUSION: There is a small subgroup of patients with postencephalitic UTLE for whom the outcome is favorable. The majority of our patients had multifocal seizure onset with BTLE and ETMFE, and poor outcome after surgical treatment. PMID- 11022141 TI - Epileptic negative myoclonus and brief asymmetric tonic seizures. A supplementary sensorimotor area involvement for both negative and positive motor phenomena. AB - Epileptic negative myoclonus (ENM) is an epileptic motor dysfunction characterised by brief lapses of postural tone provoked by paroxysmal cortical discharges. We report the electrophysiological and video-polygraphic findings in a patient presenting with the unusual association of ENM and brief asymmetric tonic-postural seizures of cryptogenetic origin. Focal EMG silent periods in the left deltoid muscle (mean duration 81.2 +/- 16.4 ms), time-locked with vertex spikes were present. The time lag between spikes and ENM was 33.1 +/- 4.6 ms. We suggest the involvement of mesial frontal areas in the genesis of both negative and positive motor phenomena. PMID- 11022142 TI - Ictal spitting during a left emporal lobe-originated complex partial seizure: a case report. AB - We present a 35-year-old male patient with intractable temporal lobe epilepsy in whom spitting automatism was documented by ictal recordings during seizures. Spitting is an uncommon automatism and occurs mainly with right-sided, nondominant, temporal focus. However, our patient had left mesial temporal sclerosis with nonverbal memory impairment, but intracarotid amobarbital test demonstrated language and memory dominance on the right hemisphere. The authors feel that this case supports the hypothesis of a nondominant, temporal lobe origin for the spitting automatism. PMID- 11022143 TI - Drop episodes in Coffin-Lowry syndrome: an unusual type of startle response. AB - We present a patient with a complete Coffin-Lowry syndrome, associated with drop episodes precipitated by sudden auditory stimuli, which provoked in turn, a definite loss of muscle tone in both legs. Electrophysiological studies showed that these episodes are an unusual type of startle response and that they may be associated with Coffin-Lowry syndrome. PMID- 11022144 TI - Rotatory seizures are not so rare as described. Reply to the letter of Saka and Saygi, Vol. 2, No 2, June 2000. PMID- 11022146 TI - [Fetal weight gain curve for a cohort of 126 pregnant women at Ouagadougou, Burkino Faso]. AB - We carried out a prospective study at Ouagadougou, from March 1 1997 to April 30 1998, in which we plotted curves of fetal weight gain, estimated by ultrasound biometry, with the aim of comparing our results to those of other authors and establishing a local chart. We established a growth curve from 936 fetal weights estimated during the follow up of 126 pregnant women. The mean age of the women included was 26 +/- 0,8 years and 52.4% were of the mean socioeconomic level. We identified significant differences between our weight gain curve and those of other authors. The factors involved in this difference are unclear but may be nutritive, ecological and ethnic in nature. Further work is required to produce local charts, to optimize the estimation of fetal weight. PMID- 11022145 TI - [Determinants of dental caries in Haitian schoolchildren and implications for public health]. AB - This work was carried out as part of a community development project in North West Haiti. The aim was to determine the prevalence of dental caries among young Haitians and to identify the principal determinants and risk factors. A total of 322 schoolchildren, all aged 12 years, participated in the study in March and April 1996. Demographic characteristics, hygiene and dietary habits were recorded. In parallel, a dentist assessed the amount of debris present on the teeth and investigated the children's history of caries. Almost two thirds of the children examined were free of caries. The mean number of decayed missing and filled teeth (DMFT index) was 0.93, which is low. However, a high-risk group was identified with a mean DMFT index of 3.8. This group is a source of some concern, particularly as the region's health services are often inaccessible resulting in dental caries frequently remaining untreated until tooth extraction is required. Logistic regression analysis showed that dental hygiene and, to a lesser extent, sugar intake, were the principal risk factors for dental caries. Adolescents who consume more than three meals per day and who, presumably, have a higher intake of cariogenic food, present more caries than those who consume less. These schoolchildren meet the objectives of the WHO for dental caries for 2000, but two courses of action are nonetheless necessary: caries prevention and improved access to dental health care. To prevent caries, techniques for increasing the resistance of teeth could be recommended, as in industrialized countries. However, such strategies would be difficult to implement in the rural context of a developing country. The addition of fluoride to drinking water, for example, is not practical as most of the inhabitants of rural areas do not have access to running water. The use of sealing agents, which protect against decay affecting the occlusal surfaces of molars, is also impractical, for financial reasons. Thus, basic measures involving the improvement of dental hygiene and reducing the intake of cariogenic foods remain the principal means of preventing caries. Increasing the accessibility of dental care, like prevention, involves a number of problems. Increasing the number of health centers, or renovating existing clinics requires resources, and possibly infrastructure, that the Haitian state may not be in a position to provide. The recently developed ART (Atraumatic Restoration Treatment) overcomes this problem. This method involves manually cleaning cavities and sealing then with glass ionomer. This product is highly adhesive, which frees dentists from the constraints of having to use rotating electrical equipment. Caries can therefore now be treated in remote areas with a minimum of dental equipment. However, although encouraging results were obtained in a three-year trial in Thailand, the mechanical quality of glass ionomer and its ability to block caries development are unclear. In conclusion, a lack of financial resources limits the possibilities of intervention in developing countries, drawing attention to the important question of the financing of health programs. Funding remains the key to any program and has been shown to be an essential issue in epidemiological studies. PMID- 11022148 TI - [The current situation with regard to nicotine addiction in Burkina Faso: tobacco supply and data from the KAB-P survey of young people in Ouagadougou]. AB - Smoking is a worldwide phenomenon and many studies have demonstrated that tobacco use is a risk factor for morbidity and mortality. We investigated aspects of the supply of, and demand for, tobacco in Burkina Faso, with a view to helping the public authorities design strategies to combat this phenomenon. We used data on the importation of tobacco and the making of cigarettes in Burkina Faso and carried out a survey of adolescents and young adults (n = 289) with a mean age of 20.9 years. In 1997, Burkina Faso imported 1,905, 214 metric tons of tobacco of all kinds, with a CAF value of 5,808, 905,269 CFA francs. In the survey, 30.7% of those interviewed said that they smoked. These individuals gave various reasons for their use of tobacco including imitating friends (38.6%) and personal liberty (14.1%). These young people were generally aware of the risks of tobacco use, but this did not deter them from smoking. In countries like Burkina Faso, which is faced with many challenges, controlling tobacco advertising and providing ways and means to help young people to stop smoking or to deter them from starting to smoke are of major importance for future public health. PMID- 11022147 TI - [Ocular traumatism in children at Laquintinie Hospital, Douala (Cameroon)]. AB - Pediatric ophthalmology is poorly developed in Cameroon. However, efforts are being made to collect data essential for the development of national strategies to combat blindness. We relate our experience, within this framework, at a large public hospital in Cameroon. We carried out a retrospective study covering the period from June 1993 to May 1998, studying the medical files of children under the age of 15 years with ocular traumatism. Data were collected from 144 files. The frequency of ocular/orbital injury was 7.8% and was the third most common condition, after ametropia and kerato-conjunctivitis, in this population. The mean age of the children was 7 years and 3 months and more boys (64%) than girls were affected. Ocular lesions were due to contusion in 83.3% of cases. The principal causes of the trauma were children's games (40.2%), and punishment by parents or teachers (23.7%). The ocular lesions were similar to those described in previous studies. Infection was rare, because the interval from trauma to consultation was very short. The functional prognosis was severe, with 24.3% of patients having final vision less than 1/10. We recommend that children should be informed about the dangers of violent games and that parents and teachers should be made aware of the dangers associated with brutality towards children. Finally, ophthalmologists should ensure that the injured eye is treated rapidly and carefully so as to minimize functional sequelae. PMID- 11022149 TI - [Congenital malaria. Parasitological and serological studies in Niamey (Niger)]. AB - Congenital malaria is defined as the presence of Plasmodium parasites in the erythrocytes of newborns less than seven days old. The aim of this study was to determine the incidence of congenital malaria and its possible clinical consequences. We carried out a prospective survey in Niamey, the capital of Niger (600,000 inhabitants) from July to September 1993. Niamey is in an area of mesoendemic malaria and this period of the year corresponds to the rainy season, when malaria transmission is maximal. Ninety mothers and their newborns were included. We assessed the clinical status of the mother and child at the time of the delivery, and took blood smears to check for parasitemia and blood samples to check for antimalaria antibodies by indirect immunofluorescence (IIF). The placenta was not examined. Clinical signs of malaria (fever, splenomegaly, anemia and jaundice) were absent in all mothers and children and 88 of the 90 children had normal birth weights. Plasmodium falciparum was the only parasite detected, with 49 of the 90 mothers and 12 of the 90 newborns having positive blood smears. Serological tests detected the presence of antimalaria antibodies in 73 of the 90 mothers (81.1%) and 68 of the 90 newborns (75.5%). Thus, we found no cases of congenital malaria with clinical signs in this study, despite the high frequency of parasites and antimalaria antibodies. The reasons for this absence of cases of congenital malaria with symptoms are discussed. PMID- 11022150 TI - [Prevention of infection in a surgical environment in the regional hospitals of Senegal]. AB - This study evaluated the attitudes and practices of the personnel with respect to the prevention of nosocomial infections in a hospital environment. We carried out a qualitative survey of all categories of personnel between July 1998 and March 1999, at the five regional hospitals in Senegal (Thies, Kaolack, Saint-Louis, Diourbel, and Louga). Data were collected in two ways: using a questionnaire to evaluate knowledge and by observing the attitudes and practices of the personnel. Hands were rarely washed before and after each procedure and surgical washing was often performed in poor conditions. The decontamination of soiled equipment was ineffective. Reusable instruments were washed directly with bare hands or by individuals wearing used surgical gloves. The equipment used for sterilization was dilapidated and unsuitable and the norms for sterilization were seldom respected. Circulation in the region of the operating theatre was disorganized, if not anarchic. The personnel had a high risk of contamination from blood. Biomedical waste was not decontaminated: it was collected poorly and disposed of directly into the environment. None of the sites visited had a program of waste incineration. In conclusion, at the sites visited, there is a high risk of nosocomial contamination during care and the reuse of equipment, for both the staff and the patients treated. PMID- 11022151 TI - [Teenage knowledge about sex]. AB - The aim of this study was to investigate the quality and sources of teenagers' information about sex. Between October 26th and December 16th 1997, 277 pupils from lower and upper schools agreed to participate in this study. Two hundred and seventy (97.5%) of the pupils in this population declared that they had received information about sex. This information was provided principally by the media, followed by friends, boyfriends and girlfriends, the school and their parents. The information received concerned sexually transmitted diseases (STDs and AIDS), the consequences of sexual relationships, sexual hygiene and the signs of puberty. We found that 95.5% of the pupils knew about AIDS and gonorrhea and that 91.7% knew how to protect themselves against AIDS and STDs. Thus, most pupils had received information, from various sources, on sexuality. Parents played little role in the sexual education of their children, and the State was involved in various ways. PMID- 11022152 TI - [Sensory heterotropias: epidemiological aspects at Douala General Hospital (Cameroon)]. AB - The aim of this retrospective study was to determine the epidemiological characteristics of the sensory heterotropia cases seen in the Opthamology Department of Douala General Hospital between November 1991 and November 1999. Sensory heterotropia accounted for 22.5% of the cases of strabismus reported. Onset was before the age of one year in 80.4% of cases and after the age of one year in 19.6% of cases. All cases of esotropia were apparent before the age of one year. Sensory heterotropia was associated with amblyopia in 89.3% of cases, with eccentric fixation in 78.6% of cases and with nystagmus in 53.6% of cases. Etiology was diverse, the most frequent causes being: albinism (19.6%), chorioretinal scars (16.1%), unilateral cataracts (16.1%), atrophy of the optic nerve (16.1%) and ametropia (8.9%). We recommend preventive measures for some etiologies. PMID- 11022153 TI - [Typhoid and paratyphoid fever in adults in the Internal Medicine Department at Libreville (Gabon)]. AB - We carried out a retrospective study of 150 files of patients suffering from typhoid or paratyphoid fever, confirmed bacteriologically, between January 1992 and December 1996 at Libreville. Young adults were the most frequently affected. These infections were associated with unstable living conditions and lack of cleanliness. Neurological (17%), digestive (12%) and cardiovascular (3%) complications were the principal factors aggravating the disease. The following serotypes were identified: Salmonella typhi (47%), S. paratyphi B (25%), S. paratyphi C (20%) and S. paratyphi A (8%). Salmonellosis was associated with schistosomiasis (13%), nematodiasis (2.3%), sickle-cell anemia (7%) and HIV infection (8%). The mortality rate was 3.3%. Fluoroquinolones are the molecules of choice for treating typhoid and paratyphoid fevers. PMID- 11022154 TI - [Immunosuppression and vaccinations]. AB - Immunocompromised subjects have a higher risk of infection. Some infectious risks could be controlled by vaccination, carried out according to normal schedules, in the absence of effective curative treatment. According to the type of immunodeficiency and its severity, certain vaccines may be recommended, others should be avoided and still others may be used with no particular risk. Thus, in immuno-compromised subjects, vaccines consisting of inactivated, inert or dead microorganisms are indicated. In contrast, those consisting of live microbes are not recommended for several reasons: 1) there is a risk that the immunocompromised host will be unable to control infection with the vaccine; 2) there is a risk of an increase in viral replication, particularly in HIV-infected subjects and 3) there is a risk that the response of the immune system will be poor. HIV-infected individuals now account for most of the cases of secondary immuno-deficiency, following the emergence of AIDS in 1980. They are increasing in number, especially in developing countries, where antiretroviral treatment is not widely available. In this context, vaccinations against transmissible childhood viral and bacterial diseases (e.g. measles and polio) with a high prevalence is advisable, with modifications according to the risk of contagion and the degree of immunodeficiency. However, these vaccines do not target opportunist infections, the prognosis of which is poor and against which we have no vaccines. Other vaccines are recommended only for immunocompromised subjects visiting countries with specific epidemiological situations (endemic or epidemic diseases) or if vaccination is required by the country visited. PMID- 11022156 TI - Finally--a positive change from IPS to PPS! PMID- 11022157 TI - Policy implications of medication errors. PMID- 11022155 TI - [Malaria in the central health district of Dakar (Senegal). Entomological, parasitological and clinical data]. AB - We previously investigated malaria in the southern health district of the city of Dakar, which includes the oldest neighborhoods. In this study, we investigated malaria in the central health district, corresponding to the central area of the conurbation. The study was carried out at 12 sites, from March 1996 to February 1997. The sites were selected such that the entire district was covered and included 2 sites in the shanty town and three in an old village that has been absorbed into the city. We carried out prospective monthly entomological analyses with a view to identifying the vectors and the mode of transmission of malaria. We also carried out clinical and parasitological follow up to determine the incidence of parasitemia and of bouts of malaria. Insects were collected overnight from humans and the insects remaining the next morning in 10 bedrooms in the health district were collected. For clinical and parasitological follow up, families were visited at home once per week and their clinical state was assessed. Blood smears were taken to facilitate the detection of bouts of malaria. Body temperature was measured and we checked for the presence of organisms in the blood systematically during the last weekly visit of each month. For a total of 308 collections at night from human volunteers and 1,395 bedroom collections of residual fauna, we obtained 12,879 Culicidae females, 199 (1.5%) of which were anopheles mosquitoes, with Culex quinquefasciatus accounting for 98% of the remaining mosquitoes. As in the southern district, A. arabiensis was the only species of the A. gambiae complex collected. Anopheles mosquitoes accounted for only 0.3 bites per man per night and 0.07 females per room. They were therefore poorly represented in this district and were not detected at all at five sites. They were found in large numbers only during the rainy season, especially in September, when they accounted for 2.25 bites per man per night and 0.3 females per room at 3 sites in an undeveloped zone in which 81.4% of all the anopheles mosquitoes were collected. The parturition frequency of the biting females was 32.6% and that of the females collected in houses was 50.0%. None of the A. arabiensis females dissected (98.5% of those collected) carried Plasmodium sporozoites. The clinical and parasitological follow up concerned 2,583 individuals, aged from 1 month to 80 years, from 285 families resident in Dakar who volunteered for the study; 41.9% of these individuals were less than 15 years old and 92.2% had been living in Dakar for more than 2 years. Thick and thin blood smears taken monthly showed the frequency of the parasite to be 1.0% and that of gametocytes to be 0.1%. P. falciparum was the only parasite detected in the subjects. Plasmodium infections were observed in all age groups, with a frequency of 0.4% (adults over the age of 20 years) to 1.6% (children under two years of age). Parasitized subjects were detected in every month of the study, with a frequency of 0.4% (in January) to 1.9% (in December). The largest number of cases detected in a three-month period (38.8% of all cases) was that for October to December, the three-month period immediately after the rainy season (July to September). Parasite frequency, which was no higher than 1. 2% at 10 sites, was clearly higher at two sites in the shanty town (3.8 and 6.8%), mostly inhabited by immigrants from rural areas. At the end of the study year, satisfactory weekly follow up was considered to have been achieved for 1,067 of the participants. The annual incidence of parasitemia in this cohort was 5.1% and that of malaria was 2.4%. Incidence did not vary significantly with age and was between 1.8% and 7.6% for parasitemia and between 0.8% and 3.5% for malaria. However, significant differences in incidence were observed between areas. Incidence was higher at the two sites in the shanty town, with rates of 12.1% and 36.5% for parasitemia and 6.1% and 15.9% for malaria. (ABSTRACT TR PMID- 11022158 TI - The gaze of nursing in home care: a commentary. PMID- 11022159 TI - Heart failure. PMID- 11022160 TI - Case management: more important than ever with prospective payment. PMID- 11022161 TI - Industry urges Congress to restore access to home care. PMID- 11022162 TI - Psychopharmacology: an update for psychiatric home care. AB - Psychopharmacology is a critical component of psychiatric home care. Nurses play an important role in medication management for patients who require this intervention. Knowledge of psychopharmacologic assessment, phases of psychopharmacologic treatment, psychobiology, and critical issues related to specific classes of medication is essential in developing a treatment plan with the patient and implementing it. Recent psychobiology research has indicated the importance of neurotransmitters, genetics, and the effect of the immune and endocrine systems. Critical issues related to anxiolytic medications, mood stabilizers, antidepressants, and antipsychotic (neuroleptic) medications are described. PMID- 11022163 TI - Pediatric pain assessment. AB - Pain in children is frequently underrecognized or undertreated based on misconceptions and misjudgments about the experience of pain in this age group. The goal of pain assessment is to obtain sufficient and accurate data, depending on the developmental stage of the child. This information can come from questioning the child directly, including using pain rating scales, and evaluating physiological and behavioral changes. Assessing the emotional state of the family and caregivers is also crucial. Health care providers need to be persistent and thorough when evaluating pain in children and use this information to formulate and modify an appropriate treatment plan. PMID- 11022164 TI - Exploring the clinical pharmacist's role in improving home care for patients with diabetes. AB - Diabetes is a common condition among clients receiving home care services. Improved glycemic control can greatly thwart the long-term complications of diabetes. A study examining the role of a clinical pharmacist was performed in a home care agency in which 105 clients were interviewed and their respective charts reviewed with the following results: 37% were incorrectly classified as to type of diabetes, serum creatinine and HbA1c values were not commonly recorded, and clients tended to have complex drug regimens with a large number of potential drug-related problems. The clinical pharmacist's potential impact in this environment is great based on the data gathered from this study. PMID- 11022165 TI - Deliberations on the risks and antibiotic management of surgical infections: preface PMID- 11022166 TI - Risk factors and surveillance for surgical wound infections. PMID- 11022167 TI - Current trends in antibiotic prophylaxis in surgery. PMID- 11022168 TI - Diagnosis and treatment of uncomplicated and complicated surgical infections. PMID- 11022169 TI - Human leptin: from an adipocyte hormone to an endocrine mediator. AB - Leptin is a mainly adipocyte-secreted protein that was discovered 5 years ago. Most of the research following this discovery focused on the role of leptin in body weight regulation, aiming to illuminate the pathophysiology of human obesity. However, more and more data are emerging that leptin is not only important in the regulation of food intake and energy balance, but that it also has a function as a metabolic and neuroendocrine hormone. It is now clear that it is especially involved in glucose metabolism, as well as in normal sexual maturation and reproduction. Besides this, interactions with the hypothalamic pituitary-adrenal, thyroid and GH axes and even with haematopoiesis and the immune system have also been described. It has been shown that leptin secretion by the adipocyte is partly regulated by other hormones, such as insulin, cortisol, and sex steroids, mainly testosterone. Also, other hormones like thyroid hormone and GH are possibly involved in leptin synthesis. Leptin itself exerts effects on different endocrine axes, mainly on the hypothalamic-pituitary gonadal axis and on insulin metabolism, but also on the hypothalamic-pituitary adrenal, thyroid and GH axes. Leptin may thus be considered a new endocrine mediator, besides its obvious role in body weight regulation. PMID- 11022171 TI - Pygmy mouse gene mutation protects against obesity. PMID- 11022170 TI - Cushing's syndrome: from patients to proteins. PMID- 11022172 TI - Increased sensitivity to the inhibitory effect of excess iodide on thyroid function in patients with beta-thalassemia major and iron overload and the subsequent development of hypothyroidism. AB - OBJECTIVE: Patients with beta-thalassemia frequently develop primary hypothyroidism and other endocrine disorders due to iron overload. We studied whether administration of excess iodide to patients with apparently normal thyroid function could uncover an underlying thyroid disease. DESIGN AND METHODS: Twenty-five patients, 10 prepubertal (mean age 11+/-3 years) and 15 adults (mean age 23+/-5 years) with normal thyroid hormone and TSH levels, a normal response of TSH to TRH and negative thyroid peroxidase antibodies received 20mg iodide three times daily for three weeks, and thyroid hormone and TSH levels were measured weekly during, and for three weeks after, iodide administration and every 3 months thereafter for the next 5 years. RESULTS: During iodide administration there was a significant decrease in thyroid hormone concentrations which remained within normal levels, and a significant increase in TSH concentrations which in 14 out of 25 (56%) patients reached the hypothyroid level. Baseline TSH values were higher in those patients who developed subclinical hypothyroidism (2.31+/-0.71mU/l vs 1. 34+/-0.64mU/l, P=0.0016). Subclinical hypothyroidism developed in 70% of prepubertal and in 47% of adult patients. Serum ferritin was elevated in all patients. Nine of the fourteen patients (64.3%) who developed subclinical hypothyroidism during iodide administration developed hypothyroidism during the 5-year follow-up compared with only one of the eleven patients with a normal response to iodide (P=0.004). CONCLUSIONS: Patients with beta-thalassemia should not be exposed to excess iodide due to increased sensitivity to its inhibitory effects on thyroid function. The susceptible individuals frequently develop permanent hypothyroidism in the following years. PMID- 11022173 TI - Power spectral analysis of heart rate in hypothyroidism. AB - OBJECTIVE: The aim of the present study was to evaluate the impact of hypothyroidism on the autonomic regulation of the cardiovascular system by analysing separately sympathetic and parasympathetic influences on the heart. DESIGN: In seven newly diagnosed untreated hypothyroid patients we analysed power spectral density of heart rate cyclic variations at rest, while lying, and while standing. The same protocol was repeated after the induction of stable euthyroidism by levothyroxine (L-T(4)) treatment. The results were also compared with those obtained from seven age-, sex- and body mass index-matched control subjects. METHODS: Heart rate variability was evaluated by autoregressive power spectral analysis (PSA). This method allows reliable quantification of low frequency (LF) and high frequency (HF) components of the heart rate power spectral density. These are considered to be under mainly sympathetic and purely parasympathetic control respectively. In addition, heart rate variations during deep breathing, lying to standing, and Valsalva's manoeuvre were assessed. RESULTS: PSA showed a sharp reduction in the HF (parasympathetic) component in hypothyroid subjects compared with controls (lying, 29.4+/-5.4 vs 47.7+/-6.3 normalized units (NU) (means +/- s.e.m.), P<0.05; standing, 14.0+/-3.5 vs 32.1+/ 3.6NU, P<0.005). Conversely, the LF (mainly sympathetic) component was higher in hypothyroid subjects than in controls (lying, 61.6+/-6.4 vs 45.4+/-6.7 NU; standing, 71.7+/-8.0 vs 53.1+/-5.6NU), this difference being significant in the standing position. Hence, the LF/HF ratio, which is considered an index of sympathovagal balance, was increased in hypothyroid subjects while both lying (2.75+/-0.6 vs 1.16+/-0.3; P<0.05) and standing (10.0+/-3.7 vs 1.85+/-0.3; P<0. 02). Total heart rate variability, expressed as total power spectral density, was lower in hypothyroid patients than in control subjects, this difference being significant in the lying position (574+/-126 vs 2302+/-994ms(2), P<0.05). In patients re-examined after L-T(4) therapy, complete normalization of cardiovascular parameters was observed (LF/HF ratio, lying, 1.26+/-0.4; standing, 2.56+/-0.8, both P<0.01 vs baseline values). The response to conventional cardiovascular autonomic tests was not significantly different between hypothyroid patients and healthy controls, and did not change in patients after therapy. CONCLUSIONS: These results suggest that, contrary to the clinical picture, thyroid hormone deficiency is associated with an increased sympathetic influence on the autonomic cardiovascular system. The changes in sympathetic function could be explained by a secondary adaptation to an altered cardiovascular responsiveness. PMID- 11022174 TI - Medullary thyroid carcinoma: an accurate pre-operative diagnosis by reverse transcription-PCR. AB - OBJECTIVE: To study the expression of calcitonin (CT) and thyroglobulin mRNA in samples of leftover cells in needles used for fine-needle aspiration biopsy either from thyroid tumours or cervical lymph nodes. PATIENTS AND METHODS: Specimens were analysed using reverse transcription-polymerase chain reaction; 12 samples from 11 patients were included and molecular diagnosis was compared with cytological or histological diagnosis and serum CT measurements. RESULTS: Transcripts of the CT gene were detected in all six patients with medullary thyroid carcinoma (MTC) but in none of the other patients. CONCLUSIONS: Present data reinforce this technique as a reliable and alternative tool to establish the pre-operative diagnosis of MTC, especially when cytological examination is not conclusive or when cytological information is not in agreement with clinical data. Furthermore, it may be clinically useful to identify those conditions in which increased serum CT in the presence of a thyroid nodule is not due to MTC. PMID- 11022175 TI - The usefulness of conventional and echo colour Doppler sonography in the differential diagnosis of toxic multinodular goitres. AB - OBJECTIVE: To assess the potential role of conventional sonography and colour flow Doppler (CFD) sonography (CFDS) in the differential diagnosis of toxic multinodular goitres. SUBJECTS AND METHODS: We investigated 55 patients with untreated hyperthyroidism (24 with typical toxic diffuse goitre of Graves' disease (Group A); 26 with multinodular goitre (Group B); and five with single toxic adenoma (Group C); 22 euthyroid subjects (12 with non-toxic multinodular goitre (Group D) and ten normal subjects (Group E)) were included as controls. In all cases free thyroxine, free tri-iodothyronine, TSH, TSH receptor antibodies (TRAb), anti-thyroperoxidase antibody, anti-thyroglobulin antibodies and anti thyroid microsomal antibodies were determined and a [(99m)Tc]pertechnetate thyroid scan was performed. RESULTS: Patients with toxic multinodular goitre displayed two different CFDS patterns: 18 patients (Group B-1) had nodules with normal vascularity surrounded by diffuse parenchymal hypoechogenicity with markedly increased CFD signal and maximal peak systolic velocity (PSV) (a pattern similar to Group A patients with Graves' disease); eight patients (Group B-2) had increased intra- and perinodular CFD signal and PSV with normal extranodular vascularity (a pattern similar to that found in Group C patients with single toxic adenoma). Patients of Group B-1 showed a proportion of clinically evident thyroid ophthalmopathy, positive TRAb and other thyroid autoantibodies similar to that observed in Group A patients, while no evidence of thyroid autoimmunity was found in Group B-2. Sixteen out of 18 (89%) patients from Group B-1 displayed a scintiscan pattern of diffuse uneven radionuclide distribution, while seven out of eight (87.5%) of those from Group B-2 had localized uptake in multiple discrete nodules. Taken together, these data strongly suggest that Group B-1 mostly represents patients with the multinodular variant of Graves' disease, while Group B-2 represents patients with non-autoimmune toxic multinodular goitre. CONCLUSIONS: This study shows that combined conventional sonography and CFDS may easily distinguish nodular variants of Graves' disease from non autoimmune forms of toxic multinodular goitre and confirms the clinical usefulness of this technique in the first-line evaluation of hyperthyroid patients. PMID- 11022176 TI - Central hypocortisolism as part of combined pituitary hormone deficiency due to mutations of PROP-1 gene. AB - BACKGROUND: One of the causes of combined pituitary hormone deficiency (CPHD) is represented by Prophet of Pit-1 (PROP-1) gene inactivating mutations. This disorder is generally characterized by GH, TSH, prolactin (PRL), and gonadotropin deficiency, but recent papers have described a concomitant alteration of the corticotrope function. OBJECTIVE: To make a detailed investigation of the hypothalamic-pituitary-adrenal axis in two sisters with PROP-1 gene mutations. PATIENTS: Two female siblings (17 and 16 years old) with CPHD, belonging to a Brazilian family of consanguineous parents, presented with growth retardation and central hypothyroidism during childhood, and showed central hypogonadism at the age of puberty. No clear clinical symptoms and signs of hypocortisolism were present. METHODS: GH, TSH, free thyroxine, total tri-iodothyronine, PRL, LH, FSH, ACTH and cortisol were measured in basal condition and after appropriate testing. The molecular study was performed by PCR amplification and sequencing analysis of PROP-1 gene. RESULTS: Both patients showed GH, PRL, LH and FSH deficiencies, associated with absent responses to an insulin tolerance test (ITT), TRH and GnRH injection. Circulating concentrations of TSH were normal in basal conditions, but failed to respond to a TRH test. Plasma ACTH concentrations were normal, but serum cortisol concentrations were below the lower limit of the normal range, showing a trend to decrease during 6 years of follow-up. The serum ACTH response to ITT was impaired, whereas its response to CRH was normal and prolonged. The cortisol response to both tests, and to the ACTH test, was clearly impaired. In both sisters, the genetic analysis showed the presence of a homozygous 2-bp deletion (296delGA) of PROP-1 gene, which results in the synthesis of a protein with no residual functional activity. CONCLUSION: In addition to GH, TSH, PRL and gonadotropin deficiency, patients with PROP-1 gene mutations can present with late-onset central hypocortisolism, possibly beause of the lack of important paracrine factors normally produced by the cells surrounding the corticotropes and absent in the pituitary of these patients, or because of progressive corticotrope apoptosis. This finding indicates the need for life-long endocrine monitoring of PROP-1-deficient patients. PMID- 11022177 TI - Occurrence and effects of octreotide antibodies during nasal, subcutaneous and slow release intramuscular treatment. AB - OBJECTIVE: Previous studies have indicated that antibody formation against octreotide is extremely rare. We examined the occurrence of octreotide antibody formation after treatment with three administration forms in large populations of patients with acromegaly or carcinoid syndrome. DESIGN: (i) Nasally administered octreotide: 70 previously untreated patients and 81 previously s.c. octreotide treated patients participated. (ii) Subcutaneously administered octreotide: 172 acromegalic patients and 59 patients with carcinoid syndrome treated for up to 12 years participated. (iii) Intramuscularly administered depot octreotide (Sandostatin LAR): 62 acromegalic patients participated. METHODS: Presence of antibodies is defined as increased precipitation by polyethylene glycol of (125)I octreotide after incubation with serum; this was also used for screening of cross reaction with somatostatin and lanreotide (Somatuline). RESULTS: In patients who received nasal octreotide for at least 9 and up to 12 months (n=42), the occurrence of octreotide antibodies was 77% and 81% for previously untreated and treated patients respectively. In subcutaneously treated patients it was 63/231 (27%) after a mean exposure of 3 years. In patients treated for more than 5 years (n=53) it was 57% and after 8 years (n=18) 72%. In contrast, no patient could with certainty be identified to be antibody-positive after a mean of 2.5 years intramuscular Sandostatin LAR treatment (n=47). In all populations, the antibody positive patients were as well controlled as the antibody-negative patients. Octreotide antibodies did not cross-react with native somatostatin (n=141), while about 25% of the antibody-positive sera did cross-react with the somatostatin analogue, lanreotide (Somatuline, Ipstyl, Angiopeptin). CONCLUSIONS: Antibody formation against octreotide is much more frequent than previously believed. It depends primarily on drug exposure time and route of administration. It does not alter the GH/IGF-I status in treated acromegalic patients and induces only mild local reactions in some patients. PMID- 11022178 TI - Cardiac dysfunction in acromegaly: evidence by pulsed wave tissue Doppler imaging. AB - OBJECTIVE: To verify whether the accuracy of data on myocardial function provided by pulsed-wave tissue Doppler imaging (PWTDI), a new echocardiographic application that allows quantitative measurements of myocardial wall velocities, could help towards a better understanding of the natural history of acromegalic cardiomyopathy. DESIGN: Eighteen patients with active acromegaly (ten men and eight women; mean age 48.0+/-15.0 years) with no other detectable cause of heart disease underwent PWTDI. Thirteen healthy individuals matched for age and body mass index acted as a control group. METHODS: Ejection fraction (EF), transmitral early/late diastolic velocity (E/A) ratio and isovolumic relaxation time (IVRT) were measured by conventional echocardiography; systolic peak (Sv) and early (Ev) and late (Av) diastolic peak velocities, Ev/Av ratio and regional IVRT (IVRTs) were obtained by PWTDI. RESULTS: All patients showed appreciably abnormal left ventricular global diastolic function represented by prolongation of the IVRT (P<0.001). Using PWTDI we found a prolongation of IVRTs and inversion of the Ev/Av ratio. In addition, the Ev/Av ratio proved to be significantly negatively correlated with IVRT; this correlation was not present in the case of the E/A ratio. Furthermore, a decrease in Sv was detected in the basal segment of the lateral wall (P<0.01), which had the greatest degree of diastolic dysfunction. CONCLUSIONS: PWTDI confirmed the acknowledged diastolic dysfunction that accompanies acromegalic cardiomyopathy and highlighted the greater sensitivity of regional PWTDI with respect to global Doppler diastolic indexes. Furthermore, by revealing an impairment of regional systolic function in presence of a normal EF, the findings with PWTDI contradicted the largely accepted theory that systolic function remains normal for several years in patients affected by acromegalic cardiomyopathy. PMID- 11022179 TI - Acute effects of interferon-alpha administration on testosterone concentrations in healthy men. AB - OBJECTIVE: Recombinant human interferon alpha (rhIFN-alpha) is used therapeutically in malignant disorders and chronic hepatitis. The present study was assessed to study the effects of rhIFN-alpha on the hypothalamic-pituitary testicular (HPT) axis. DESIGN AND METHODS: We performed a saline-controlled cross over study in six healthy men, sequentially measuring the serum concentrations of gonadotropins, testosterone, the free androgen index (FAI) and sex hormone binding globulin (SHBG) after a bolus subcutaneous injection of rhIFN-alpha. RESULTS: rhIFN-alpha induced a sustained decrease of both testosterone (from 19.5+/-1.88 to a nadir of 5.49+/-0.51nmol/l at the end of the study) and FAI (from 98.7+/-14.7 to a nadir of 32. 1+/-5.3 at the end of the study), whereas concentrations of LH, FSH and SHBG were not different between the two studies. CONCLUSIONS: Our results suggest that rhIFN-alpha affects the HPT axis at the testicular level, either directly or indirectly, and changes feedback relationships between the pituitary and the testis. PMID- 11022180 TI - Pituitary response to luteinizing hormone-releasing hormone in women with variant luteinizing hormone. AB - OBJECTIVE: To assess the LH response of the pituitary gland to GnRH stimulation in healthy women with a mutant beta-subunit (Trp8 to Arg8 and Ile15 to Thr15). DESIGN: Clinical study. PATIENTS: We studied 40 healthy non-pregnant Japanese women of known zygosity for the LH beta-subunit gene (3 homozygotes for the mutant gene, 17 heterozygotes, and 20 homozygotes for the wild type). All women had normal ovulatory cycles. MEASUREMENTS: Serum LH status was determined by comparing LH immunoassays results using a monoclonal antibody recognizing only wild-type LH with those from a polyclonal antibody assay recognizing both variant and wild-type LH. The ratio of monoclonal to polyclonal immunoassay results determined the serum LH status. LH secretion in response to a GnRH stimulation test was measured. RESULTS: All women with the wild-type LH showed a normal response of LH to GnRH according to both assays. Over the time course of the response, the ratios in women with wild-type LH showed no remarkable changes. The response curves in women heterozygous for the mutant peaked 15-30min after GnRH injection; their response patterns included a statistically significant decrease in the rates of response at 15min after injection. CONCLUSIONS: There are the differences in circulatory kinetics between the two LH forms and in regulation of the two types of LHbeta genes. The maximal response of the variant LH to pituitary stimulation with GnRH appears to be greater than that of wild-type LH. PMID- 11022181 TI - Ovarian-adrenal cross-talk in polycystic ovary syndrome: evidence from wedge resection. AB - OBJECTIVE: To determine whether the ovary influences adrenal androgen secretion in women with polycystic ovary syndrome (PCOS). DESIGN: Six PCOS-affected patients with clomiphene resistance and gonadotrophin hyperresponsivity, and six controls with regular ovulatory cycles, matched for age and body mass index. METHODS: Bilateral ovarian wedge resection was performed to induce ovulation surgically for these refractory women with PCOS. The adrenal androgen secretions were evaluated in PCOS patients before and again 6 months after this surgery, and in the controls, using an ACTH stimulation test (0.25mg synthetic ACTH(1-24)). RESULTS: Biochemically, basal levels and the maximum net increases (Delta) of 17 hydroxyprogesterone (17-OHP) and androstenedione, Delta17-OHP/Delta progesterone and Delta androstenedione/Delta17-OHP ratios in response to exogenous ACTH were significantly higher in PCOS patients before operation than those of controls. This purely ovarian surgery in women with PCOS was found to significantly reduce their basal androstenedione, testosterone and LH levels, insulin/glucose ratio, and post-corticotrophic Delta17-OHP, Delta androstenedione, Delta17-OHP/Delta progesterone and Delta androstenedione/Delta17-OHP, without obvious changes in FSH, oestradiol, sex hormone-binding globulin, Delta dehydroepiandrosterone, Delta dehydroepiandrosterone sulphate, Delta aldosterone and Delta cortisol values. CONCLUSIONS: Ovarian hyperandrogenicity from polycystic ovary may contribute to the enhanced adrenal P450c17alpha activity and subsequent Delta(4) androgen reserve revealed by the pharmacological corticotrophin stimulation in our special PCOS cases. PMID- 11022182 TI - The nicotinic acid analogue acipimox increases plasma leptin and decreases free fatty acids in type 2 diabetic patients. AB - The effect of 3 days of intensive treatment with acipimox, an antilipolytic nicotinic acid derivative, on plasma leptin levels was studied in eight patients with Type 2 diabetes mellitus in a double-blind, placebo-controlled, cross-over study. Acipimox reduced plasma free fatty acids (FFA) markedly and lowered plasma triglycerides, glucose and insulin. Plasma leptin levels were elevated in all eight patients during 3 days of acipimox treatment (mean increase+/-s.e.: 2.38+/ 0.57ng/ml, P<0.005) and the 24h mean effect of acipimox on leptin levels increased during the experimental period (P<0.03). The effect on plasma insulin and glucose resembled a mirror image of the effect on plasma leptin during 3 days of treatment. The suggestion that leptin mediates insulin resistance and may be involved in the development of the diabetic syndrome cannot be supported by the present results. It has been reported that FFA stimulates leptin secretion. Surprisingly, despite a markedly reduced FFA level, leptin concentration increased in the present study. We suggest that a primary acipimox effect is to increase leptin secretion, and that this prevails over the reduced FFA stimulus. PMID- 11022183 TI - Genotype-phenotype associations in non-classical steroid 21-hydroxylase deficiency. AB - OBJECTIVE: To evaluate whether genotype differences can explain the clinical variability of non-classical steroid 21-hydroxylase deficiency (NC21-OHD) and to determine if genotype is related to ethnic origin. DESIGN: Genotyping for mutations in the steroid 21-hydroxylase (CYP21) gene was performed in 45 unrelated Israeli Jewish patients (nine males) with NC21-OHD (60min 17 hydroxyprogesterone (17-OHP), 45-386nmol/l) who were referred for evaluation of postnatal virilization or true precocious/early puberty. Eleven siblings diagnosed through family screening were genotyped as well. METHODS: Patients were divided by genotype into three groups: (A) homozygous or compound heterozygous for the mild mutations (V281L or P30L) (n=29; eight males); (B) compound heterozygous for one mild and one severe mutation (Q318X, I2 splice, I172N) (n=12; no males); (C) mild mutation detected on one allele only (n=4; one male; peak 17-OHP 58-151nmol/l). We then related the genotype to the ethnic origin, clinical phenotype and hormone level. Since group C was very small, comparisons were made between groups A and B only. RESULTS: At diagnosis, group B tended to be younger (5. 8+/-3.0 vs 8.1+/-4.3 years, P=0.09), had greater height SDS adjusted for mid-parental height SDS (1.6+/-1.1 vs 0.7+/-1.4, P=0.034), tended to have more advanced bone age SDS (2.9+/-1.5 vs 1.7+/-2.1, P=0.10) and had a higher peak 17-OHP level in response to ACTH stimulation (226+/-92 vs 126+/-62nmol/l, P<0.01). Group B also had pubarche and gonadarche at an earlier age (5.1+/-2.4 vs 7.4+/-2.2 years, P<0.01 and 7.4+/-1.8 vs 9.9+/-1.4 years, P<0.001, respectively) and a higher rate of precocious puberty (50 vs 17%, P=0.04). Stepwise logistic regression analysis (excluding males) yielded age at gonadarche as the most significant variable differentiating the two groups, with a positive predictive value of 86% for a cut-off of 7.5 years. CONCLUSIONS: The findings suggest that genotype might explain some of the variability in the phenotypic expression of NC21-OHD. Compound heterozygotes for one mild and one severe mutation have a higher peak 17-OHP associated with pubarche and gonadarche at an earlier age and more frequent precocious puberty. Hence, the severity of the enzymatic defect might determine the timing and pattern of puberty. PMID- 11022184 TI - The best correlation of the new index of hyperandrogenism with the grade of increased body hair. AB - OBJECTIVE: Hyperandrogenemia is the most frequent endocrine disorder in fertile women causing a variety of negative metabolic disturbances. Establishing the diagnosis of androgen overproduction has important implications for the follow-up and treatment of patients. The aim of our study was to identify the optimal laboratory marker of androgen production by correlating the markers to the presence or grade of increased body hair as a clinical sign of hyperandrogenism. DESIGN: Prospective observational study. METHODS: A total of 62 women with acne were included into the study. The serum concentrations of testosterone, androstenedione, dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS) and sex hormone-binding globulin (SHBG) were evaluated. The index of free testosterone (IFT) and a new index of hyperandrogenism (IHA) were calculated. The monitored laboratory markers were correlated to the presence or grade of increased body hair using several statistical methods. RESULTS: The statistical significance of differences between the average levels of laboratory markers between hirsute and non-hirsute women decreased in the following order: IHA, androstenedione and DHEA. Of all the above laboratory markers, only increased IHA was present significantly more often in hirsute women. The significance of correlation between the grade of increased body hair and the tested variables decreased in the following order: IHA, IFT, DHEA, androstenedione, DHEAS and testosterone. CONCLUSIONS: The clinical marker of hyperandrogenism correlates most closely to IHA, reflecting the levels of all commonly determined androgens or androgen precursors and SHBG. Its simple calculation makes IHA a suitable tool for determining total production of androgens in clinical practice, especially in cases with borderline elevations of values. PMID- 11022185 TI - Growth hormone and mild exercise in combination increases markedly muscle mass and tetanic tension in old rats. AB - OBJECTIVE: A decline of skeletal muscle mass and strength is seen with aging and immobilization. Growth hormone (GH) has been shown to increase muscle mass. In the present study the effects of a combination of mild exercise and GH on skeletal musculature tetanic tension, dry defatted weight (DDW), volume, water, fat and collagen concentrations were investigated in old rats. DESIGN: Recombinant human GH (2.7mg/kg per day) was injected subcutaneously for 73 days in 21-month-old female rats. Exercised rats ran on a treadmill, 8 m/min for 1 h/day. The in vivo maximal tetanic tension of the calf musculature (m. soleus, m. plantaris, m. gastrocnemius together) was analysed in anaesthetized rats by stimulating the ischiadic nerve. RESULTS: The maximal tetanic tension was increased by 23% in GH-injected compared to saline-injected rats. Mild exercise + GH in combination resulted in a further 18% increase in maximal tetanic tension. The mild exercise by itself did not influence the maximal tetanic tension significantly when compared with saline injected rats. The GH administration and/or mild exercise did not change skeletal muscle endurance, measured as tetanic tension during 30s of stimulation. Serum IGF-I concentration was increased twofold in GH-injected rats. CONCLUSION: The increased muscle mass induced by GH + mild exercise was associated with a corresponding increase in maximal tetanic tension. Combination of GH + mild exercise resulted in a substantial further increase of muscle mass and maximal tension compared with GH injections alone in these old rats. PMID- 11022186 TI - Endocrine activities of alexamorelin (Ala-His-d-2-methyl-Trp-Ala-Trp-d-Phe-Lys NH2), a synthetic GH secretagogue, in humans. AB - OBJECTIVE: Peptidyl and non-peptidyl synthetic GH secretagogues (GHS) possess significant GH-, prolactin (PRL)- and ACTH/cortisol-releasing activity after i.v. and even p.o. administration, acting via specific hypothalamo-pituitary receptors in both animals and humans. The hexapeptide hexarelin (HEX) is a paradigmatic GHS whose activities have been widely studied in humans. The heptapeptide Ala-His-d-2 methyl-Trp-Ala-Trp-d-Phe-Lys-NH(2) (alexamorelin, ALEX) is a new synthetic molecule which inhibits GHS binding in vitro, but its endocrine activity has never been studied in humans. DESIGN: In six young adults we studied the effects of 1.0 and 2.0 microgram/kg i.v. ALEX or HEX on GH, PRL, ACTH, cortisol and aldosterone levels and those of 20mg p.o. ( approximately 300 microgram/kg) on GH levels. RESULTS: Basal GH, PRL, ACTH, cortisol and aldosterone levels in all testing sessions were similar. ALEX and HEX (1.0 and 2.0 microgram/kg i.v.) induced the same dose-dependent increase of GH and PRL levels. Both ALEX and HEX induced a dose-dependent increase of ACTH and cortisol levels. The ACTH and cortisol responses to the highest ALEX dose were significantly higher than those after HEX. Aldosterone levels significantly increased after both i.v. ALEX doses, but not after HEX. The GH response to 20mg p.o. ALEX was higher, though not significantly, than that to the same HEX dose. CONCLUSION: ALEX, a new GHS, shows the same GH-releasing activity as HEX. On the other hand, ALEX seems endowed with an ACTH-releasing activity more marked than that of HEX; this evidence could explain the significant increase of aldosterone levels after its i.v. administration. PMID- 11022187 TI - In vitro study of the effect of raloxifene on lipid metabolism compared with tamoxifen. AB - OBJECTIVE: Tamoxifen and raloxifene, selective estrogen receptor modulators, decrease serum concentrations of total cholesterol; however, the effect of these drugs on triglyceride metabolism is unclear. In the present study, we investigated the in vitro effect of raloxifene on lipid metabolism and compared it with that of tamoxifen. DESIGN AND METHODS: Intracellular concentrations of total cholesterol and triglyceride in HepG2 cells were measured by an enzymatic method after tamoxifen or raloxifene treatment with or without oleic acid and with or without very low density lipoprotein. RESULTS: Intracellular concentrations of total cholesterol and triglyceride without oleic acid or very low density lipoprotein were not significantly different after treatment with tamoxifen or raloxifene. In contrast, although raloxifene with oleic acid did not increase the intracellular concentrations of triglyceride, tamoxifen treatment in the presence of oleic acid or very low density lipoprotein significantly increased (P<0.05) the triglyceride concentrations. CONCLUSION: The present study suggests that raloxifene does not increase intracellular triglyceride in the presence of oleic acid or very low density lipoprotein, in contrast to tamoxifen. Therefore, raloxifene might be safer than tamoxifen for treating patients with unstable triglyceride levels or a history of hypertriglyceridemia. PMID- 11022188 TI - In vivo and in vitro suppression by leptin of glucose-stimulated insulin hypersecretion in high glucose-fed rats. AB - OBJECTIVES: Chronic feeding to rats of high glycaemic index (GI) diets results in the hypersecretion of insulin in response to an i.v. glucose load. The first aim of this study was to see if this exaggerated insulin response was accompanied by a hypersensitivity to glucose stimulation in isolated islets in vitro. The second aim was to see if the adipocyte factor, leptin, was able to alter insulin secretion in this model both in vivo and in vitro. DESIGN AND METHODS: Rats were fed for 6 weeks either a high GI diet in which the carbohydrate component was mostly glucose (GLUC diet) or a low GI diet containing mostly amylose (AMOSE diet). Rats then underwent an i.v. glucose tolerance test (ivGTT) (1g/kg) with and without a prior infusion of leptin (133 microg/kg perh). Islets were then isolated from these rats and basal and glucose-stimulated insulin secretion (GSIS) measured in both the absence and presence (100ng/ml) of leptin. RESULTS AND CONCLUSIONS: Peak insulin response during the ivGTT was 3-fold greater in GLUC rats (P<0.001). Leptin had no effect on AMOSE rat insulin response but lowered the GLUC rat response to AMOSE rat levels. In vitro, basal insulin secretion was 4-fold greater in GLUC rats (P<0.05). At 20mmol/l glucose, there was no further increase in insulin secretion in GLUC rats but a 2-fold increase in AMOSE rats. Leptin had no effect on basal insulin secretion or GSIS in AMOSE rats but reduced basal insulin secretion and GSIS in GLUC rats. These results show insulin hypersecretion in high GI-fed rats may be reduced by leptin. PMID- 11022190 TI - New perspectives for gene therapy in endocrinology. AB - Gene therapy for endocrine diseases represents an exciting new type of molecular intervention that may be a curative one. Endocrine disorders that might be treated by gene therapy include monogenic diseases, such as GH deficiency and hypothalamic diabetes insipidus, and multifactorial diseases, such as diabetes mellitus, obesity and cancer. Premises seem promising for endocrine tumours, but many combined approaches of cell and gene therapy are foreseeable also for other endocrine disorders. This review outlines the principles of gene therapy, describes the endocrine disorders that might take advantage of gene transfer approaches, as well as the gene therapy interventions that have already been attempted, their major limitations and the problems that remain to be solved. PMID- 11022189 TI - Catecholamines suppress leptin release from in vitro differentiated subcutaneous human adipocytes in primary culture via beta1- and beta2-adrenergic receptors. AB - OBJECTIVE: Circulating leptin, the product of the ob gene, is known to be closely correlated with adipose tissue mass, but it is also subject to short-term regulation by a variety of hormones including catecholamines. The aim of this study was to investigate the contribution of the three beta-adrenergic receptors to leptin secretion from cultured human adipocytes. DESIGN AND METHODS: The model of in vitro differentiated human subcutaneous adipocytes was used in this study. The presence of the beta-adrenoceptor subtypes was studied by RT-PCR. The functional role of the receptor subtypes was determined by stimulation of lipolysis by selective beta-adrenergic agonists and by measuring glycerol release. Leptin secretion into the medium of cultured human adipocytes from young normal-weight females was measured by radioimmunoassay. RESULTS AND CONCLUSION: In a first set of experiments, the expression of the three beta-adrenergic receptor subtypes in cultured human adipocytes was demonstrated. To test their functional activity, the effect of the beta-adrenoceptor agonists isoproterenol (non-selective agonist), dobutamine (beta(1)-selective), fenoterol (beta(2) selective) and the beta(3)-selective agonists BRL 37344 and CGP 12177 was studied. All agonists exhibited a dose- and time-dependent stimulation of glycerol release into the medium in a rather uniform manner. Isoproterenol rapidly reduced leptin secretion from cultured subcutaneous adipocytes in a dose dependent fashion. Incubation with 10(-6)mol/l isoproterenol for 24h resulted in a reduction of the leptin concentration by 48% (P < 0.01). A similar, but less pronounced suppressing effect was seen for dobutamine and fenoterol, whereas both BRL 37344 and CGP 12177 were not effective. These data provide evidence that catecholamines are able to suppress leptin release from differentiated human adipocytes, supporting the concept that leptin secretion is acutely regulated by surrounding hormones. This inhibition is obviously mediated via beta(1)- and beta(2)-adrenergic receptors. PMID- 11022191 TI - Identification of follistatin as a possible trait-causing gene in polycystic ovary syndrome. PMID- 11022192 TI - A novel activating mutation in the thyrotropin receptor gene in an autonomously functioning thyroid nodule developed by a Japanese patient. AB - OBJECTIVE: A number of activating mutations of the thyrotropin receptor (TSHR) have been found in autonomously functioning thyroid nodules (AFTNs) in European patients. We aimed to study TSHR mutation in AFTNs in Japanese patients because no TSHR activating mutation has been found by previous incomplete studies. DESIGN: A typical AFTN developed in a 69-year-old Japanese woman was studied. METHODS: The entire exon 10 of the TSHR cDNA was sequenced. Functional studies were done by site-directed mutagenesis and transfection of a mutant construct into COS-7 cells. RESULTS: We identified a novel heterozygous TSHR gene mutation, Leu512-->Arg (L512R; CTG-->CCG), from the AFTN. The mutation was not detected in the adjacent normal thyroid tissue. COS-7 cells transfected with L512R mutant TSHR expression vector exhibited a 3.3-fold increase in basal cAMP level compared with that of cells transfected with wild-type TSHR DNA, confirming that the mutation was the direct cause of the AFTN. TSHR activating mutations involving the third transmembrane helix reported to date are S505R/N and V509A as well as L512R. An in vitro site-directed mutagenesis study encompassing residues 505-513 revealed that mutations involving residues other than these three did not show constitutive activation. CONCLUSION: This is the first TSHR activating mutation found in a Japanese patient, although true prevalence of TSHR activating mutations in AFTNs developed in Japanese patients remains to be elucidated. In addition, functional studies suggested that amino acid residues in the third transmembrane helix maintaining inactive conformation of the TSHR seem to be located on the same surface of the alpha-helix, possibly making interhelical bonds with another helix. PMID- 11022193 TI - Prevalence of iodine deficiency and goitre during pregnancy in east Hungary. AB - OBJECTIVE: To assess the iodine nutritional status and the prevalence of goitre during pregnancy in a region of Hungary that appeared to be iodine sufficient in previous studies. DESIGN: A cross-sectional voluntary screening study was organized in which 313 pregnant women participated. METHODS: Urine iodine concentration and the volume of the thyroid gland were measured in every woman. In the presence of low urinary iodine concentrations, goitre, or both, thyroid function tests were performed. RESULTS: Iodine deficiency was found in 57.1% of the pregnant women, and was severe in 15.6%. The volume of the thyroid gland was enlarged in 19.2% of individuals. Nodular goitre was found in 17 women (5.4%). The frequency of goitre and the mean thyroid volume were increased in the group of iodine-deficient women. In the 89 cases of iodine deficiency or goitre, thyrotrophin concentrations were in the normal range; however, the free triiodothyronine:free throxine ratio was increased in 97% of them, indicating that the thyroid gland was in a stimulated state in these individuals. CONCLUSIONS: Iodine deficiency with high prevalence of goitre was recognized among pregnant women in an area that previously appeared to be iodine sufficient. An unexpected mild iodine deficiency was also noted in the non-pregnant control group. Reassessment and continuous monitoring of iodine nutritional status is warranted even in populations that are apparently considered to be 'at no risk' of iodine deficiency, especially in pregnant women. Regular administration of iodine, starting at preconception or in early pregnancy and continuing during the period of nursing, is recommended in these regions. PMID- 11022194 TI - Comparative study of thyroid function and types of thyroid dysfunction in two areas in Denmark with slightly different iodine status. AB - OBJECTIVE: The pattern of thyroid dysfunction seems to depend on the iodine status of the population. Prevalence of thyroid dysfunction could be a parameter to consider when evaluating iodine deficiency disorders in a population. DESIGN: Comparative cross-sectional investigation in two regions in Denmark with marginally different iodine excretion. METHODS: A random selection of 4649 participants from the Civil Registration System in Denmark in age groups between 18 and 65 years were examined. Thyroid dysfunction was evaluated from blood samples and questionnaires, and compared with results from ultrasonography. RESULTS: Median iodine excretion was 53 microg/l in Aalborg and 68 microg/l in Copenhagen. Previously diagnosed thyroid dysfunction was found with the same prevalence in the regions. Serum TSH was lower in Aalborg than in Copenhagen (P=0. 003) and declined with age in Aalborg, but not in Copenhagen. Not previously diagnosed hyperthyroidism was found with the same overall prevalence in the regions, but in age >40 years hyperthyroidism was more prevalent in Aalborg (1.3 vs 0.5%, P=0.017). Not previously diagnosed hypothyroidism was found more frequently in Aalborg (0.6 vs 0.2%, P=0.03). Hyperthyroidism was more often associated with macronodular thyroid structure at ultrasound in Aalborg and hypothyroidism was more often associated with patchy thyroid structure in Copenhagen. CONCLUSIONS: Significant differences in thyroid dysfunction were found between the regions with a minor difference in iodine excretion. The findings are in agreement with a higher prevalence of thyroid autonomy among the elderly in the most iodine-deficient region. PMID- 11022195 TI - Long-term follow-up evaluation of magnetic resonance imaging in the prognosis of permanent GH deficiency. AB - OBJECTIVE: In patients with GH deficiency (GHD), magnetic resonance imaging (MRI) has revealed morphological abnormalities such as pituitary hypoplasia, pituitary stalk agenesis (PSA) and ectopia of the posterior pituitary (PPE). The MRI anomalies have been more frequently reported in patients with multiple pituitary hormone deficiency (MPHD) than in subjects with isolated GH deficiency (IGHD). The aim of this work was to define which MRI anatomical abnormalities of the hypothalamo-pituitary area can be considered as a prognostic marker of permanent GHD. DESIGN: To investigate the relationship between the neuroradiological images and endocrine findings, we clinically re-evaluated 93 out of the 121 GHD patients with IGHD and MPHD previously studied. RESULTS: No additional hormone deficiencies were observed in 55 out of 60 patients initially classified as having IGHD with a normal (15 cases) or reduced (40 cases) pituitary gland size, without other MRI abnormalities. The remaining five children, who had initially shown an apparently IGHD in spite of PSA and PPE, developed a MPHD over time. In 33 MPHD patients with (25 cases) or without (8 cases) MRI abnormalities, the associated hormone deficiencies were confirmed during follow-up. CONCLUSIONS: The IGHD patients showing PSA and PPE inevitably develop additional hormone deficiencies, while IGHD subjects having no MRI abnormalities maintain IGHD. Moreover, the anatomical abnormalities of the hypothalamo-pituitary area can be considered as a prognostic marker of permanent GHD. PMID- 11022196 TI - Organ-specific and non-organ-specific autoantibodies in children and young adults with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). AB - OBJECTIVE: The aim of the study was to assess the complex of autoantibodies which can be detected in patients with autoimmune polyendocrinopathy-candidiasis ectodermal dystrophy (APECED), a rare autosomal recessive disease in which the extent of autoimmunity is still unknown. DESIGN: Antibodies (A) to parathyroid glands, adrenal cortex (AC-A), ovary and testis (steroid cell antibodies, SC-A), pancreatic islet cells (IC-A), gastric parietal cells, and non-organ-specific antigens were investigated in 11 APECED patients living in the Salento region of southern Italy. Further measurements included antibodies to cytochrome P450 (CYP) enzymes: cholesterol side-chain cleavage enzyme (CYP11A), 21-hydroxylase (CYP21) and 17alpha-hydroxylase (CYP17); and to glutamic acid decarboxylase 65-kDa isoform (GAD65), tyrosine phosphatase-like protein IA2, thyroglobulin (TG), thyroperoxidase (TPO), thyrotropin receptor, liver CYP enzymes and intrinsic factor. METHODS: Antibodies to organs and subcellular fractions were detected by immunofluorescence. Radiobinding, immunoradiometric, and immunoblotting assays were used for the other measurements. RESULTS: AC-A and SC-A were positive in all sera; among antibodies to adrenal CYP enzymes, only CYP21-A were present in all the patients with Addison's disease of short-medium duration (<15 years). Of three patients with Addison's disease of long duration (>15 years), two tested positive for antibodies to all three CYP enzymes, and the other for only CYP11A A. In all sera CYP11A-A and/or CYP17-A were found. Two patients tested positive for both IC-A and GAD65-A, one for both IC-A and IA2-A, and one for GAD65-A; the fasting C-peptide assay showed no statistical difference between these four subjects and the others. All four hypothyroid patients were positive for TPO-A, while two of them were positive and two were negative for TG-A; two euthyroid subjects had positivity for TG-A. Liver-kidney microsomal antibodies reacting against the CYP2A6 were detected in two patients with autoimmune hepatitis. All but one sera contained anti-nuclear antibodies at a titre ranging between 1:20 and 1:80; however, only two patients had a connective tissue disease (Sjogren's syndrome). CONCLUSIONS: Several autoantibodies may be detected in any APECED patient. Our data confirm that CYP21-A and TPO-A are major autoantibodies involved in APECED-associated Addison's disease and hypothyroidism respectively, while CYP11A-A and CYP17-A correlate with positivity for SC-A. Markers of islet cell autoimmunity are frequent, but prevalence and modalities of progression to overt beta-cell failure have to be clarified. Low-titre non-organ-specific autoantibodies are a feature of autoimmunity in APECED, but their role has yet to be fully explained. PMID- 11022197 TI - Circulating IGF-I concentrations are low and not correlated to glycaemic control in adults with type 1 diabetes. AB - OBJECTIVE: To study plasma concentrations of insulin-like growth factor-I (IGF-I) in adults with type 1 diabetes (IDDM) in comparison with a reference population, and the influence of glycaemic control, dose of insulin, and sex on the concentration of circulating IGF-I in IDDM. DESIGN AND METHODS: Patients with type 1 diabetes were recruited consecutively from our outpatient diabetes unit. In all, 79 men and 55 women aged 20-60 years with a disease duration >/=6 years (range 6-51 years) took part in the study. A reference population of 80 men and 83 women aged 20-60 years was randomly obtained from the population registry. IGF I was measured with radioimmunoassay after acid-ethanol extraction. RESULTS: Mean +/- s. d. values of IGF-I were lower in patients with diabetes (146+/-66 microg/l) than in controls (238+/-83 microg/l, P<0.001). Those with diabetes had lower IGF-I concentrations in all age groups and the differences were highly significant in all decades except in women aged 50-59 years. IGF-I was negatively correlated with age in patients and controls. No correlation was found between IGF-I and glycaemic control measured as haemoglobin A(1c) (HbA(1c)) in the patients. IGF-I was positively associated with the dose of insulin/kg body weight in male patients independently of age, HbA(1c) and body mass index (P<0.03), but not in female patients (P=0.14). CONCLUSIONS: Our data show that IGF-I concentrations are low in adult patients with type 1 diabetes with a disease duration >/=6 years, independently of glycaemic control. This suggests that subcutaneous insulin substitution is inadequate to normalize circulating IGF-I concentrations in patients without endogenous insulin secretion. PMID- 11022198 TI - Mutations in the insulin promoter factor-1 gene in late-onset type 2 diabetes mellitus. AB - OBJECTIVE: Insulin promoter factor-1 (IPF-1) is a transcription factor expressed in pancreatic beta cells. Following the identification of missense variants in the coding regions of the IPF-1 gene, in subjects selected for a strong family history of type 2 diabetes, the aim of our study was to evaluate the prevalence of these variants in the common form of type 2 diabetes. METHODS: Three variants (C18R, Q59L and D76N) were screened by PCR-RFLP in a group of 296 unrelated French late-onset type 2 diabetic subjects consecutively recruited in a diabetes department of a university hospital, regardless of family history of diabetes. RESULTS: The C18R and Q59L variants were each found in 0.37% of the diabetic patients, and in none of 147 controls. We did not detect the D76N variant, which was the most frequent variant in subjects with a strong family history of diabetes, in patients or controls. CONCLUSIONS: We have observed a combined prevalence of missense variants in the coding region of the IPF-1 gene of around 1%, in unselected patients with the common form of late-onset type 2 diabetes. The prevalence of these variants in subjects with a strong family history of type 2 diabetes had been found to be as high as approximately 6%. These differences in prevalence might be related to differences in the clinical profile of patients, such as age of onset of diabetes and associated obesity, as well as a family history of diabetes. PMID- 11022199 TI - The highly malignant phenotype of anaplastic thyroid carcinoma cell lines is recessive. AB - OBJECTIVE: The aim of our studies was to determine whether the phenotype of the anaplastic thyroid carcinomas is dominant or recessive. In fact, it is hypothesized, on the basis of epidemiological and pathological data, that undifferentiated thyroid carcinomas are derived from differentiated tumours through a mechanism of tumour progression. DESIGN: Cell hybrids have been generated by cell fusion of anaplastic thyroid carcinoma cell lines, which show a highly malignant phenotype, to cell lines deriving from differentiated thyroid carcinoma, which show a non-tumorigenic or a poorly tumorigenic phenotype. All of the parental cell lines showed impaired p53 gene function. RESULTS: The cell hybrids contained alleles from the parental cell lines. All of the cell hybrids showed a lower growth rate compared with the parental undifferentiated carcinoma cell lines and were unable to grow in soft agar and to induce tumours after injection into athymic mice. CONCLUSION: Taken together, these findings suggest that the highly malignant phenotype of the anaplastic thyroid carcinoma is achieved by the impairment of gene functions that negatively regulate cell growth, rather than by the activation of dominant oncogenes. PMID- 11022200 TI - Evaluation for roles of nitric oxide generated in the anteroventral third ventricular region in controlling vasopressin secretion and cardiovascular system of conscious rats. AB - OBJECTIVE: To examine local actions of nitric oxide (NO) on the neural mechanisms controlling the release of vasopressin (AVP) and the cardiovascular system in the anteroventral third ventricular region (AV3V), a pivotal area for autonomic functions, and to pursue the problem of whether it may have any role in the AVP and cardiovascular responses evoked by plasma hypertonicity or by increased prostaglandin E(2) (PGE(2)) in the AV3V - one possible factor implicated in osmotic responses. METHODS: We infused NO-related agents into the AV3V, its adjacent area, the nucleus of the vertical limb of the diagonal band (VDB), or into the lateral cerebral ventricle of conscious rats, monitoring effects on plasma AVP, osmolality, sodium, potassium and chloride, arterial pressure and heart rate in the presence or absence of an osmotic or PGE(2) stimulus. The infusion sites were determined histologically. RESULTS: Infusion of L-arginine, the substrate of NO synthase (NOS), into the AV3V structures such as the median preoptic nucleus and periventricular nucleus produced dose-related increases in plasma AVP, arterial pressure and heart rate 5 or 15 min later, whereas infusion of D-arginine (which is not a substrate for NO synthesis) was without significant effect on these variables. Plasma osmolality or electrolytes were not changed by these treatments. The AV3V infusion of sodium nitroprusside (SNP), a spontaneous releaser of NO, also induced dose-dependent augmentations of plasma AVP, without evoking remarkable alteration in the cardiovascular parameters. The infusion of L or D-arginine into the VDB affected none of the variables significantly. When applied intracerebroventricularly, L-arginine caused only increases in plasma AVP, whereas SNP caused only reductions in arterial pressure, leaving other variables at stable values. The effects of AV3V L-arginine on plasma AVP and the cardiovascular variables were abolished by N(G)-nitro-L-arginine methyl ester (L NAME), a potent inhibitor of NOS, applied 15 min before. In contrast, infusion of L-NAME to the AV3V did not exert a significant effect on the responses of plasma AVP or cardiovascular variables to AV3V application of PGE(2) or i.v. infusion of hypertonic NaCl. The infusion of L-NAME alone did not affect plasma variables including AVP, although it tended to increase basal arterial pressure and heart rate. CONCLUSION: These results suggest that NO generated in or near the AV3V may act to enhance AVP release, arterial pressure and heart rate, but it may not play an essential role in eliciting the responses of these variables to osmotic or PGE(2) stimuli. PMID- 11022201 TI - Development of obesity in transgenic rats with low circulating growth hormone levels: involvement of leptin resistance. AB - BACKGROUND: Human growth hormone (hGH) transgenic (TG) rats have been produced in our laboratory. These TG rats are characterized by low circulating hGH levels, virtually no endogenous rGH secretion, and massive obesity. OBJECTIVE: To elucidate how energy balance and leptin sensitivity contributed to the establishment of this obesity. DESIGN AND METHODS: Food intake, locomotor activity and leptin concentrations in serum and cerebrospinal fluid were measured in TG rats and their non-transgenic littermates (control). The effect of intraperitoneal and intracerebroventricular injection of leptin on food intake and body weight gain was also examined. RESULTS: An increase in food intake and a decrease in locomotor activity were observed from 4 and 7 weeks of age, respectively, in the transgenic rats compared with control. Serum leptin concentrations of the transgenic rats were more than twice as high as those of control rats and were associated with an increased white adipose tissue mass and ob gene expression. Intraperitoneal injection of leptin significantly decreased food intake and body weight gain in control rats, but not in transgenic rats. Leptin concentration in the cerebrospinal fluid of transgenic rats was not different from that of control rats, and intracerebroventricular injection of leptin was similarly effective in reducing food intake and body weight gain as it was in control rats. CONCLUSIONS: These results suggest that the transgenic rats, whose GH secretion is suppressed, develop obesity due to early onset of an increase in food intake and a decrease in locomotor activity with leptin resistance resulting from deteriorating leptin transport from peripheral blood to cerebrospinal fluid. PMID- 11022202 TI - Effects of a new steroidal aromatase inhibitor, TZA-2237, and/or chlormadinone acetate on hormone-induced and spontaneous canine benign prostatic hyperplasia. AB - OBJECTIVE: It has been known for many years that human benign prostatic hyperplasia (BPH) is composed predominantly of hyperplastic stromal cells rather than epithelial cells. In the present study the effects of a new steroidal aromatase inhibitor on hormone-induced and spontaneous canine BPH were investigated. METHODS: (1) Effects of TZA-2237 on hormone-induced canine BPH. Ten castrated beagles were administered testosterone and androstenedione 6 days/week for 8 months, and divided randomly into three groups after 2 months of treatment as follows. Group I served as controls, Group II was given 0.5 and Group III was given 2.5 mg/kg/day TZA-2237 5 days/week for 6 months. (2) Effects of TZA-2237 on spontaneous canine BPH. Twenty aged beagles with BPH were divided into five groups, Group IV was untreated, Group V was treated with 1 and Group VI with 5mg/kg/day TZA-2237 5 days/week for 31 weeks. Group VII was treated with 5mg/kg/day Atamestane and Group VIII was treated with 0.3 mg/kg/day chlormadinone acetate (CMA) 5 days/week. (3) Effects of TZA-2237 combined with CMA on spontaneous canine BPH. Three aged beagles with BPH were treated with 1mg/kg/day TZA-2237 and 0.03 mg/kg/day CMA 5 days/week for 20 weeks (Group IX) and a further three aged beagles with BPH were treated with 0.3 mg/kg/day CMA alone 5 days/week (Group X). RESULTS: Hormone-induced prostatic growth was significantly suppressed in group III compared with that in other groups. In Group III, the intraprostatic aromatase activity, estradiol level and androgen receptor content decreased significantly in comparison with the values in Group I. The prostatic weights in Groups V, VI and VII increased significantly in comparison with the weight in Group IV. Serum LH and testosterone levels in Groups V, VI, and VII increased significantly in comparison with the level in Group IV. The prostatic weight in Group IX was decreased only slightly, but the smooth muscle component was decreased significantly. CONCLUSIONS: TZA-2237 is a new, unique and effective aromatase inhibitor that causes inhibition of both epithelial and stromal compartments in hormone-induced canine BPH. Dual inhibition of androgen and estrogen resulted in inhibition of smooth muscle growth, and should prove effective as a new method of treatment given the atrophic effects on not only the epithelium but also the stroma in human BPH. PMID- 11022203 TI - Polymorphisms at the 5' end of the human gonadotropin-releasing hormone receptor gene are not associated with the timing of menarche in Japanese girls. PMID- 11022205 TI - Percutaneous coronary intervention in the elderly: a comparison of procedural and clinical outcomes between the eighth and ninth decades. AB - AIMS: To compare short-term safety and outcome of percutaneous coronary intervention (PCI) in the elderly in their ninth decade with those in their eighth decade. METHODS: Five-hundred and eighty-nine patients aged 70 years undergoing coronary angioplasty were stratified into two groups, those 80 years old (Group 1, n = 65) and those 70Eth 79 years old (Group 2, n = 524). RESULTS: Group 1 comprised more females than Group 2 (57% vs. 40%, respectively; p = 0.009), patients with hypertension (70% vs. 56%, respectively), untreated hyperlipidemia (18% vs. 7%, respectively), acute coronary syndromes (75% vs. 53%, respectively) and multi-vessel disease (63% vs. 54%, respectively). Angiographic success per lesion was similar in Groups 1 and 2 (96% vs. 99%, respectively) as was clinical success per lesion (89% vs. 89%, respectively). Groups 1 and 2 also had a similar incidence of in-hospital death (1.5% vs. 1.1%, respectively), Q wave myocardial infarction (0.0% vs. 0.6%, respectively), non-Q wave myocardial infarction (6.2% vs. 2.9%, respectively), emergency coronary artery bypass surgery (0.0% vs. 1. 3%, respectively), repeat PCI (3.0% vs. 1.7%, respectively), stroke (1.5% vs. 0.4%, respectively) and local vascular complications (4.6% vs. 4.4%, respectively). However, Group 1 had a longer in-hospital stay (4 days) than Group 2 (2 days) (p < 0.001). CONCLUSION: Our data suggest that short-term procedural and clinical outcomes after PCI are similar for patients in their eighth decade compared to those in their ninth decade. PMID- 11022206 TI - Percutaneous coronary intervention in the elderly: no longer the exception. PMID- 11022207 TI - Clinical outcomes following "rescue" administration of abciximab in patients undergoing percutaneous coronary angioplasty. AB - Pre-intervention administration of abciximab in patients at "high risk" for coronary angioplasty has been shown to reduce acute and long-term cardiac outcomes. The role of intra-procedural ("rescue") administration of abciximab has not been fully elucidated. We assessed the clinical outcomes associated with rescue administration of abciximab during complex percutaneous coronary interventions. We studied in-hospital and long-term (1-year) outcomes (death, myocardial infarction and target lesion revascularization) of 298 consecutive patients (78% male; age, 62 +/- 11 years; 83% with acute coronary syndrome) treated with abciximab for thrombus-containing lesions, sub-optimal angioplasty results, procedural dissections or other complications. Stents were used in 73% of procedures. Procedural success was 97.0% and overall major in-hospital complication rate was 3.0% (death, 1.3%; Q-wave myocardial infarction, 0.7%; and emergent bypass surgery, 1.0%). Most frequent angiographic complications included visible thrombus (17%), dissections (17%), threatened closure (7%), and distal embolization (7%). In-hospital non-Q wave myocardial infarction (defined as CK-MB 5 times normal) occurred in 31.0%. Out-of-hospital to one-year events included death (1.7%), Q-wave myocardial infarction (2.7%), and target lesion revascularization (15.1%); cardiac event-free survival was 82.9%. We conclude that rescue administration of abciximab is associated with relatively low in hospital complications and favorable long-term outcome in patients with sub optimal angioplasty results and/or procedure-related complications, although peri procedural non-Q wave myocardial infarction rate is high. A clinical and cost effective comparison between provisional and rescue administration of abciximab may be warranted. PMID- 11022208 TI - "Rescue" platelet GP IIb/IIIa inhibitor therapy in percutaneous coronary intervention: bailing out of a sunken ship. PMID- 11022209 TI - Contemporary percutaneous treatment of saphenous vein graft stenosis: immediate and late outcomes. AB - PURPOSE: The aim of this study was to evaluate the immediate and long-term outcomes following percutaneous treatment of an unselected series of saphenous vein graft (SVG) lesions. METHODS AND RESULTS: Consecutive interventions on 129 saphenous vein graft lesions in 101 patients were reviewed. Stents were implanted in 114 lesions (88%), which included the use of polytetrafluoroethylene-covered stents in 22 lesions (17%) and abciximab in 20 patients (20%). Angiographic success was achieved in 125 lesions (97%). In-hospital major adverse cardiac events (MACE) occurred in 11 patients (11%), with myocardial infarction being the most frequent event. Treatment of degenerated SVG lesions and SVG lesions with larger reference diameters correlated with the incidence of in-hospital MACE [odds ratio (OR) = 7.69 and 2.65, respectively; 95% confidence interval (CI) = 1.80Eth 32.8 and 0.99Eth 7.10, respectively)]. Clinical follow-up was achieved in all patients at 25 +/- 21 months. Successful revascularization to all three distributions of the major coronary arteries negatively correlated [relative risk (RR) = 0.43; 95% CI = 0.20Eth 0.92)], while treatment of a degenerated SVG positively correlated (RR = 1.92; 95% CI = 1.05Eth 3.51) with the occurrence of follow-up MACE. A final effective blood supply to the anterior wall and a higher left ventricular ejection fraction was found to negatively correlate with the occurrence of follow-up death (RR = 0. 20 and 0.61, respectively; 95% CI = 0.06Eth 0.60 and 0.41Eth 0.90, respectively). CONCLUSION: Treatment of SVG lesions continues to be associated with a high incidence of myocardial infarction, particularly in cases of degenerated SVG lesions. An effective blood supply to the anterior wall and a higher left ventricular ejection fraction were protective for the occurrence of death during the follow-up period. PMID- 11022210 TI - Stenting for saphenous vein graft disease: have we solved the problem? PMID- 11022212 TI - Why cover a good thing? PMID- 11022211 TI - Transluminally placed endovascular grafts to exclude undesirable intracardiac vascular communications. AB - PURPOSE: In this report, we summarize our early experience using transluminally placed endovascular grafts (TPEG) to close intracardiac communications within surgically created conduits. MATERIALS AND METHODS: Three patients with surgically corrected congenital heart disease were treated in this study. All three patients had intracardiac conduits [Modified Fontan (n = 2) or Mustard procedure (n = 1)]. All three patients had intracardiac shunts causing hypoxemia. The shunts were caused by a surgically created fenestration (n =1), a leak at a surgical anastomosis (n = 1) and perforations of a pericardial baffle (n = 1). All three patients were treated percutaneously from venous side approaches. The TPEG devices were constructed of pre-expanded polytetrafluoroethylene (PTFE) sutured to a Palmaz stent. RESULTS: In the two patients with the modified Fontan procedures, the leaks were completely excluded; in the patient with the Mustard procedure, the perforations in the baffle were not completely excluded. All patients experienced immediate improvements in oxygen saturation ranging from 17Eth 26% (mean, 20.3%). The improvements in oxygen saturation were sustained with values ranging from 12Eth 22% (mean, 17.3%) at the time of clinical follow up. Clinical follow-up has ranged from 13Eth 19 months (mean, 15 months) and has demonstrated improved exercise tolerance and weight gain. Echocardiographic follow-up has shown no complications of TPEG stenoses, occlusions or migrations. There were no immediate or delayed complications associated with the TPEG devices. CONCLUSION: We report our experience using TPEG devices to exclude intracardiac communications in 3 patients with surgically corrected congenital heart disease. The preliminary results are encouraging and suggest that further investigation in this area should be done. PMID- 11022213 TI - ChoICE PT wire for recanalization of chronically occluded coronary arteries: multiple wires in one? AB - Percutaneous transluminal coronary angioplasty (PTCA) of chronic total occlusions may be technically difficult and the success rate is limited despite increasing operator experience and improvements in PTCA hardware. The number of guidewires required to cross totally occluded lesions is higher than that for stenotic lesions. The ChoICE polymer-tip (PT) wire (Boston Scientific/Scimed, Inc., Maple Grove, Minnesota) is a relatively new stainless-steel core wire with a hydrophilic-coated polyurethane tip. Though never described in the literature, we found that the distal 4 cm of the wire can be cut and reshaped according to the operator's needs. Thus, instead of reshaping a kinked tip or using another new wire, the former being time-consuming and the latter expensive, one can simply cut off the kinked tip and start again with a "new wire." As the tip is resected, the wire becomes progressively more "intermediate-like" and "standard-like." We report our experience with the ChoICE PT wire in 50 consecutive cases of chronic total occlusions. The cumulative crossing success rates were 13/50 (26%) before any resections, 24/50 (48%) after 1 resection, 41/50 (82%) after 2 resections and 42/50 (84%) after 3 resections. There were no perforations, deaths, myocardial infarctions or need for bypass surgery. Our findings suggest that successful recanalization of chronic total occlusions can be achieved with a high success rate using the ChoICE PT wire. A strategy of progressively resecting the more floppy and kinked distal end can provide multiple uses from a single wire, optimizing recanalization success and obviating the need for additional wires. PMID- 11022214 TI - Platelet receptor glycoprotein IIb/IIIa inhibition with eptifibatide in a patient with thrombocytopenia after treatment with abciximab. AB - Clinical experience suggests that patients treated with the glycoprotein (GP) IIb/IIIa inhibitor abciximab (ReoPro , Eli Lilly and Company, Indianapolis, Indiana) may be at increased risk of thrombocytopenia. This case report details the successful use of the GP IIb/IIIa inhibitor eptifibatide (Integrilin , COR Therapeutics, South San Francisco, California) in a patient who developed acute thrombocytopenia (platelet count: 67,000/mm3) approximately 10 hours after initiation of abciximab therapy. Five hours after abciximab was discontinued, platelet count returned to normal (191,000/mm3) and eptifibatide was started because of persistent electrocardiographic evidence of ischemia. The patient underwent diagnostic catheterization during eptifibatide therapy, which was administered for approximately three days. Four days after the initial course of therapy with eptifibatide was discontinued, percutaneous revascularization with adjunct eptifibatide was performed. During both courses of eptifibatide therapy, platelet counts remained in the normal range (> 100,000/mm3) and no adverse ischemic or bleeding events occurred. PMID- 11022215 TI - Isolated profound thrombocytopenia associated with clopidogrel. AB - Clopidogrel is a new oral antiplatelet agent with a structure and mechanism of action similar to ticlopidine; they both block the adenosine diphosphate receptor. Because of better side-effect profile and simpler dosing regime, clopidogrel has largely replaced ticlopidine. In fact, the excellent safety and tolerability profile of clopidogrel compares favorably with that of aspirin as documented in the CAPRIE (Clopidogrel versus Aspirin in Patients at Risk of Ischemic Events) study. After marketing, several cases of clopidogrel-associated thrombotic thrombocytopenic purpura (TTP) were reported. We report our experience with a case of isolated profound thrombocytopenia without evidence of TTP after clopidogrel administration with prompt recovery of platelet counts upon discontinuation of clopidogrel and treatment with intravenous immunoglobulin. PMID- 11022216 TI - Platelets and coronary intervention: some practical precautions. PMID- 11022217 TI - Renal artery intervention: is dominance of the femoral approach about to end? PMID- 11022218 TI - Reprogramming the cortex. PMID- 11022219 TI - Rett and ICF syndromes: methylation moves into medicine. PMID- 11022220 TI - Feverish honeybees. PMID- 11022221 TI - Do life's three domains mirror the origins of sex? PMID- 11022222 TI - Daniel Bernoulli (1738): evolution and economics under risk. PMID- 11022223 TI - Patterns of replication in the neo-sex chromosomes of Drosophila nasuta albomicans. AB - Drosophila nasuta albomicans (with 2n = 6), contains a pair of metacentric neo sex chromosomes. Phylogenetically these are products of centric fusion between ancestral sex (X, Y) chromosomes and an autosome (chromosome 3). The polytene chromosome complement of males with a neo-X- and neo-Y-chromosomes has revealed asynchrony in replication between the two arms of the neo-sex chromosomes. The arm which represents the ancestral X-chromosome is faster replicating than the arm which represents ancestral autosome. The latter arm of the neo-sex chromosome is synchronous with other autosomes of the complement. We conclude that one arm of the neo-X/Y is still mimicking the features of an autosome while the other arm has the features of a classical X/Y-chromosome. This X-autosome translocation differs from the other evolutionary X-autosome translocations known in certain species of Drosophila. PMID- 11022224 TI - Cytokeratin expression in human fetal tongue and buccal mucosa. AB - Expression of cytokeratins (CK), a subset of intermediate filament (IF) proteins in epithelia, is developmentally regulated. CK expression may also change after malignant transformation. Our earlier studies on CK expression in human oral tumours and pre-cancerous lesions have shown specific changes in CK expression. We analysed CK expression in human tongue and buccal mucosa (BM) in fetuses in the embryonic age group of 16 to 27 weeks using biochemical and immunohistochemical techniques to find out whether there is any similarity in CK expression in human oral squamous cell carcinomas (SCC) and fetal oral tissues. CK 1, 8 and 18 were detected in a majority of samples using both techniques. Our earlier studies had shown aberrant expression of CK 1 and 18 in many of the oral SCC and leukoplakias. Studies by immunohistochemistry showed that these different CK antigens were expressed in different cell layers. CK 1(2) were present in the stratified epithelial layers whereas CK 8 and 18 were restricted to glandular epithelium. Till 27 weeks of gestation, both tongue and BM expressed CK 1, 8 and 18 along with CK 6 and 16. Thus, fetal tissues showed some similarities in CK pattern with their respective SCC. PMID- 11022225 TI - Mechanism of cAMP-induced H(+)-efflux of Dictyostelium cells: a role for fatty acids. AB - Aggregating Dictyostelium cells release protons when stimulated with cAMP. To find out whether the protons are generated by acidic vesicles or in the cytosol, we permeabilized the cells and found that this did not alter the cAMP-response. Proton efflux in intact cells was inhibited by preincubation with the V-type H(+) ATPase inhibitor concanamycin A and with the plasma membrane H(+) ATPase blocker miconazole. Surprisingly, miconazole also inhibited efflux in permeabilized cells, indicating that this type of H(+) ATPase is present on intracellular vesicles as well. Vesicular acidification was inhibited by miconazole and by concanamycin A, suggesting that the acidic vesicles contain both V-type and P type H(+) ATPases. Moreover, concanamycin A and miconazole acted in concert, both in intact cells and in vesicles. The mechanism of cAMP-induced Ca2(+)-fluxes involves phospholipase A2 activity. Fatty acids circumvent the plasma membrane and stimulate vesicular Ca2(+)-efflux. Here we show that arachidonic acid elicited H(+)-efflux not only from intact cells but also from acidic vesicles. The target of regulation by arachidonic acid seemed to be the vesicular Ca2(+) release channel. PMID- 11022226 TI - Role of light in the mediation of acute effects of a single afternoon melatonin injection on steroidogenic activity of testis in the rat. AB - Young adult male rats, maintained either in an LD 12 : 12 or in continuous illumination (LL) for one week, were given a single injection of 25 microg melatonin/100 g body wt or ethanolic-saline (control) at 17.00 h. Animals from each group were sacrificed at 11.00 h on the following day. The activity of two important steroidogenic enzymes, 17 beta-hydroxysteroid dehydrogenase (17 beta HSD) and delta(5)-3 beta-hydroxysteroid dehydrogenase (delta(5)-3 beta-HSD), and serum concentrations of testosterone, were measured following highly specific and sensitive spectrophotometric techniques and RIA, respectively. A significant decrease in the activity of both the steroidogenic enzymes was noted in the testes of melatonin-treated rats maintained under normal light-dark schedules, but this response was found to be lacking in the LL rats. However, no significant changes in the level of serum testosterone were noted in either group of melatonin-treated rats from the values in respective groups of ethanolic saline administered LD and LL rats. Exposure of ethanolic saline-injected rats to continuous light also did not cause any change in the steroidogenic activity of the testis from those in LD rats. The study indicates that continuous light as such does not affect the endocrine function of testis but abolishes suppressive effects of melatonin on the steroidogenic activity of the testis in rat. PMID- 11022227 TI - Comparison of the genome sequences and the phylogenetic analyses of the GP78 and the Vellore P20778 isolates of Japanese encephalitis virus from India. AB - The nucleotide sequence of the complete genomes of two Indian isolates of Japanese encephalitis virus were compared. One of these isolates, GP78 was obtained from northern India in 1978. The other, the Vellore P20778 isolate, was obtained from southern India in 1958. There was 4.40% nucleotide sequence divergence between the two Indian isolates that resulted in a 1.86% amino acid sequence divergence. Phylogenetic analyses showed that in evolutionary terms the north Indian GP78 isolate was close to the SA14 isolate from China whereas the south Indian Vellore P20778 isolate was close to the Beijing-1 isolate, also from China. The two Indian isolates, however, appear to have evolved independently. PMID- 11022228 TI - Positive correlation between menthol content and in vitro menthol tolerance in Mentha arvensis L. cultivars. AB - Menthol is a highly valued monoterpene produced by Japanese mint (Mentha arvensis) as a natural product with wide applications in cosmetics, confectionery, flavours, beverages and therapeutics. Selection of high menthol yielding genotypes is therefore the ultimate objective of all genetic improvement programmes in Mentha arvensis. A positive correlation was observed in the present study between menthol content in oils of evaluated genotypes and the level of tolerance to externally supplied menthol of explants of these genotypes in culture medium. The easy use of this relationship as a selectable biochemical marker opens the practical applicability of large scale in vitro screening of the germplasm, clones and breeders' material for selection of elite genotypes. PMID- 11022229 TI - The ontogeny of kin recognition in tadpoles of the toad Bufo melanostictus (Anura; bufonidae). AB - The ontogeny of kin recognition and influence of social environment on the development of kin recognition behaviour was experimentally investigated in tadpoles of Bufo melanostictus that lived in aggregations and showed low larval dispersion. Embryos and tadpoles of the toad were reared as (i) kin only, (ii) with kin and non-kin (separated by a mesh screen), and (iii) in isolation. They were tested for the ability to discriminate between (i) familiar siblings and unfamiliar non-siblings, (ii) familiar siblings and familiar non-siblings and, (iii) unfamiliar siblings and unfamiliar non-siblings. All tadpoles were fed on boiled spinach before conducting trials. Preference of test tadpoles to associate near the end compartments whether empty or containing members of specific stimulus groups was assessed using a rectangular choice tank. When tested in tanks with empty end compartments, the test tadpoles showed random distribution and thus no bias for the apparatus or the procedure. In the presence of kin/non kin in the end compartments a significantly greater number of test tadpoles spent the majority of the time near familiar or unfamiliar kin rather than near familiar or unfamiliar non-kin. Kin discrimination ability persisted throughout larval development. Familiarity with siblings is not required for discriminating kin from non-kin, and kin discrimination ability is not modified following exposure to non-kin. Also, involvement of dietary cues is unlikely to be the prime mechanism of kin recognition in B. melanostictus unlike in some other anurans. PMID- 11022230 TI - Mechanisms of all-trans retinoic acid-induced differentiation of acute promyelocytic leukemia cells. AB - Retinoic acids (RA) play a key role in myeloid differentiation through their agonistic nuclear receptors (RAR alpha/RXR) to modulate the expression of target genes. In acute promyelocytic leukemia (APL) cells with rearrangement of retinoic acid receptor a (RAR alpha) (including: PML-RAR alpha, PLZF-RAR alpha, NPM-RAR alpha, NuMA- RAR alpha or STAT5b-RAR alpha) as a result of chromosomal translocations, the RA signal pathway is disrupted and myeloid differentiation is arrested at the promyelocytic stage. Pharmacologic dosage of all-trans retinoic acid (ATRA) directly modulates PML-RAR alpha and its interaction with the nuclear receptor co-repressor complex, which restores the wild-type RAR alpha/RXR regulatory pathway and induces the transcriptional expression of downstream genes. Analysing gene expression profiles in APL cells before and after ATRA treatment represents a useful approach to identify genes whose functions are involved in this new cancer treatment. A chronologically well coordinated modulation of ATRA-regulated genes has thus been revealed which seems to constitute a balanced functional network underlying decreased cellular proliferation, initiation and progression of maturation, and maintenance of cell survival before terminal differentiation. PMID- 11022231 TI - Visual and acoustic communication in non-human animals: a comparison. AB - The visual and auditory systems are two major sensory modalities employed in communication. Although communication in these two sensory modalities can serve analogous functions and evolve in response to similar selection forces, the two systems also operate under different constraints imposed by the environment and the degree to which these sensory modalities are recruited for non-communication functions. Also, the research traditions in each tend to differ, with studies of mechanisms of acoustic communication tending to take a more reductionist tack often concentrating on single signal parameters, and studies of visual communication tending to be more concerned with multivariate signal arrays in natural environments and higher level processing of such signals. Each research tradition would benefit by being more expansive in its approach. PMID- 11022232 TI - Role of polyamines and ethylene as modulators of plant senescence. AB - Under optimal conditions of growth, senescence, a terminal phase of development, sets in after a certain physiological age. It is a dynamic and closely regulated developmental process which involves an array of changes at both physiological and biochemical levels including gene expression. A large number of biotic and abiotic factors accelerate the process. Convincing evidence suggests the involvement of polyamines (PAs) and ethylene in this process. Although the biosynthetic pathways of both PAs and ethylene are interrelated, S adenosylmethionine (SAM) being a common precursor, their physiological functions are distinct and at times antagonistic, particularly during leaf and flower senescence and also during fruit ripening. This provides an effective means for regulation of their biosynthesis and also to understand the mechanism by which the balance between the two can be established for manipulating the senescence process. The present article deals with current advances in the knowledge of the interrelationship between ethylene and PAs during senescence which may open up new vistas of investigation for the future. PMID- 11022233 TI - Decapod crustacean chelipeds: an overview. AB - The structure, growth, differentiation and function of crustacean chelipeds are reviewed. In many decapod crustaceans growth of chelae is isometric with allometry level reaching unity till the puberty moult. Afterwards the same trend continues in females, while in males there is a marked spurt in the level of allometry accompanied by a sudden increase in the relative size of chelae. Subsequently they are differentiated morphologically into crusher and cutter making them heterochelous and sexually dimorphic. Of the two, the major chela is used during agonistic encounters while the minor is used for prey capture and grooming. Various biotic and abiotic factors exert a negative effect on cheliped growth. The dimorphic growth pattern of chelae can be adversely affected by factors such as parasitic infection and substrate conditions. Display patterns of chelipeds have an important role in agonistic and aggressive interactions. Of the five pairs of pereiopods, the chelae are versatile organs of offence and defence which also make them the most vulnerable for autotomy. Regeneration of the autotomized chelipeds imposes an additional energy demand called "regeneration load" on the incumbent, altering energy allocation for somatic and/or reproductive processes. Partial withdrawal of chelae leading to incomplete exuviation is reported for the first time in the laboratory and field in Macrobrachium species. PMID- 11022234 TI - Cytokines and placental macrophages in regulation of birth activity. AB - The article reviews present notions on functional activity of cytokines of the fetoplacental complex. Particular emphasis is placed on the role of these molecules in the regulation of gestation processes and in pregnancy incompetence. The mechanism of the involvement of placental macrophages and their products in gestation and delivery is discussed. PMID- 11022235 TI - Regulation of cardiac function in prepubertal rats. AB - Prepubertal period in rats is characterized by some peculiarities of the cardiac regulatory mechanisms. In 6-week-old rats atropine produced bradycardia instead of tachycardia. Similar reaction was not observed in chemically sympathectomized age-matched rats. PMID- 11022236 TI - Activity of sympathoadrenal system and myelokaryocyte death during aging in AKR/JY mice. AB - Accelerated bone marrow cell death and activation of the sympathoadrenal system were observed during aging of highly leukemic 2-7-month-old AKR/JY mice compared to that in (CBA/CaLacxAKR/JY)F1 strain. Close correlation was revealed between activity of the sympathoadrenal system and necrotic and apoptotic forms of cell death. This can promote tumor process, because maximum changes in hemopoietic cells occur during advanced stage of the disease. PMID- 11022237 TI - Inhibition of alcohol uptake in C57Bl/6 mice by splenocytes extracorporeally stimulated with vasopressin. AB - Experiments on C57Bl/6 mice kept under conditions of free choice between alcohol and water for 4 and 12 months showed that adoptive transfer of splenocytes extracorporeally stimulated with arginine vasopressin suppressed alcohol intake. PMID- 11022238 TI - Liver resistance to CCl(4)-induced injury after stimulation of macrophages with various preparations. AB - Acute toxic hepatitis in male Wistar rats was produced by single injection of 40% CCl(4) (0.2 ml per 100 g body weight in oil). Pretreatment with various immunostimulators (bacterial polysaccharides prodigiozan and salmozan; yeast polysaccharides zymosan, peptidoglycan, and mannan; and hydrolytic enzyme egg lysozyme) produced a hepatoprotective effect correlating which the stimulatory influence on macrophages and increasing in the following order: mannan or = 3 mm and negative when it was < 3 mm. The validity of the test was estimated by calculating sensitivity, specificity, and the positive and negative predictive values. RESULTS: The skin prick test was positive in 82.92% of the asthmatic children and in 10.7% of the non-asthmatic children, yielding a sensitivity of 82.9%, 95% CI 76.9; 87.7 and a specificity of 89.3%, 95% IC 84.0; 93.0. The positive predictive value was 88.5%, 95% IC 83.0; 92.5 and the negative predictive value was 83.9%, 95% IC 78.2; 88.4. CONCLUSIONS: Blomia tropicalis is an important cause of bronchial asthma among the children of the northern Camaguey region. PMID- 11022270 TI - From analgesic intolerance to analgesic induced asthma: are there some determinants? AB - BACKGROUND: Analgesic intolerance (AI) sometimes appear alone and sometimes with bronchial asthma affecting about 10% of asthmatics and sometimes before and the other times after asthma. OBJECTIVE: We investigated the possible clinical risk factors which might be affecting the transition from isolated AI to analgesic induced asthma (AIA). METHODS: A total of 344 patients admitted to Hacettepe University Hospital Adult Allergy Unit between January 1991 and March 1999 and diagnosed with AI were enrolled in this survey. Patients having AIA (group I) (n = 191) were compared with the patients having AI without asthma (group II) (n = 153). The diagnosis of AI and AIA were made by history and oral provocation tests. A standard questionnaire was filled-in for all the patients. RESULTS: The risk of AIA was increased with nasal polyp, and rhinosinusitis via OR's of 2.75 (95% CI: 1.09, 6.91), and 18.58 (95% CI: 9.86, 35.01), respectively. Having a pet, and ever smoking decreased the risk of AIA in the patients with AI via OR's of 0.53 (95% CI: 0.24, 1.17), and 0.37 (95% CI: 0.17, 0.80), respectively. The association of AIA and smoking was slightly modified by food intolerance (OR for ever smoked and food intolerance: 1.31, 95% CI: 0.40, 4.30). CONCLUSION: There may be two different phenotypes of AI with different clinical features: one developing AIA (having nasal polyp and/or rhinosinusitis, and smoking if food allergy/intolerance is present), and the other AI without asthma (having pet, and could smoke). Findings of this study should be confirmed by further investigations. PMID- 11022271 TI - Local nasal immunotherapy: efficacy and tolerability of two different administration schedules in grass pollen rhinitis. AB - BACKGROUND: Efficacy and safety of local nasal immunotherapy has been demonstrated by many placebo-controlled clinical trials. The treatment schedule consists of an induction phase at increasing dosages followed by a maintenance phase. Aim of present study has been to evaluate a new simplified treatment schedule at constant dosage. METHODS AND RESULTS: 26 grass-allergic rhinitic patients have been treated according to the new constant dosage schedule, while 15 patients in a control group were treated with conventional incremental dosage schedule. A commercial preparation of allergens incorporated into powder (Allerkin). The total cumulative dosage administered with conventional schedule resulted higher than simplified constant schedule (5,025 vs 3,000 AU). To evaluate efficacy, symptom and rescue medication scores were recorded during the spring pollen season, and a subjective overall evaluation was asked to the patients at the end of the season. The analysis of scores showed that the two treatment schedules were equivalent as regards both efficacy and safety, being side effects light and restricted to the administration site (sneezing). CONCLUSIONS: The use of a constant dosage schedules is suggested for the administration of local nasal immunotherapy, having as advantage a better simplicity. Avoidance of mistakes in dosages is considered important in a therapy which is self-administered by patients. PMID- 11022272 TI - Anaphylaxis to calcitonin. AB - BACKGROUND: The calcitonin is an hormone produced by the thyroid gland C cells. The salmon calcitonin is used in some osteomuscular diseases. There are few references of allergic reaction to this hormone. We introduce a case of a sixty years old woman with several previous episodes of rhinitis, conjunctivitis and perspiration immediately after the administration of salmon calcitonin with nasal spray or intramuscular administration (Calsynar). There were some temporal periods of good tolerance between these episodes. METHODS: Skin prick test (SPT), nasal and intramuscular challenge test with commercial salmon calcitonin (Miacalcic) were performed. Leukocyte histamine release test with salmon calcitonin and serum tryptase levels at baseline and after intramuscular challenge test were performed. RESULTS: The patient skin prick test with commercial calcitonin (Miacalcic. 50 UI/ml) was positive and negative in controls. The nasal challenge test with a calcitonin nasal spray, up to 150 UI, was negative. The intramuscular challenge test with 25 UI of Miacalcic was positive with an immediate anaphylactic reaction. Whole blood histamine release studies were negative. Serum tryptase levels after intramuscular challenge did not increase significantly with regard to the basal levels. CONCLUSION: We have introduced a case of anaphylaxis by calcitonin that suggest an IgE mediated hypersensitivity reaction. PMID- 11022273 TI - Our future lies in education. PMID- 11022274 TI - Major quarantine plans in place for Sydney 2000. PMID- 11022275 TI - Inform us, AVA. PMID- 11022276 TI - Well done on microchips! PMID- 11022277 TI - PetPEP defended. PMID- 11022278 TI - Manufacturer's steroid policy. PMID- 11022279 TI - PetPEP questions. PMID- 11022280 TI - Microchip action. PMID- 11022281 TI - Blame NSW education. PMID- 11022282 TI - Docking dispute. PMID- 11022283 TI - Systemic mycosis due to Aspergillus deflectus in a dog. AB - A 4-year-old, entire female, German Shepherd Dog was referred with a 3-month history of right foreleg lameness that partially responded to nonsteroidal anti inflammatory and antimicrobial therapy. The bitch lost weight, was polydipsic and had reduced exercise tolerance. On referral, the animal was in poor condition, pyrexic and exhibited moderate pain on full extension of the right shoulder. Blood, urine and joint fluid were obtained and radiographs were taken of the right shoulder and chest. The bitch was lymphopaenic, hyperfibrinogenaemic, hyperglobulinaemic, mildly azotaemic, mildly proteinuric and isosthenuric. Branching fungal hyphae were present in the urine. On radiography, the thorax contained a large ventral mediastinal mass and the humeral head had extensive areas of radiolucency. An aspirate from the right humeroscapular joint exhibited branched fungal hyphae and numerous neutrophils and macrophages. A diagnosis of disseminated mycosis was made and euthanasia was performed. At necropsy, numerous caseating granulomas were present, especially in the kidneys, adrenal glands, heart and lymph nodes. Extensive osteomyelitis involved the head of the right humerus, the sternebrae and the fifth intervertebral disc. Fungal hyphae were detected in sections of granulomas in all affected organs and a diagnosis of disseminated fungal granulomatosis was made. Aspergillus deflectus was readily isolated from affected lymph nodes, but confirming its identity as A deflectus using standard procedures proved difficult. The identity of the fungus was finally confirmed by sequencing part of the 185 rRNA of the isolate. This is the first report in Australia of a disseminated mycosis caused by A deflectus. Previously, the involvement of A deflectus as a cause of disseminated mycosis was limited to 5 cases from the West Coast of the USA, four of which occurred in German Shepherd Dogs. PMID- 11022284 TI - Entrapment of the small colon through a mesocolic rent in a mare. AB - A 6-year-old mare was presented for acute abdominal pain unresponsive to analgesics. Exploratory laparotomy revealed entrapment of the small colon through a 12 cm rent in the mesocolon. The incarcerated small colon was manually reduced and the rent in the mesocolon was sutured closed. The mare made excellent postoperative recovery and was discharged from the hospital 4 days later. The cause of the rent, which was chronic in appearance, is unknown. PMID- 11022285 TI - Disseminated haemangiosarcoma in an eastern barred bandicoot (Perameles gunnii). AB - A captive adult male Eastern barred bandicoot (Perameles gunnii) presented with three palpable subcutaneous masses in November 1998. A diagnosis of haemangiosarcoma was made based on histological examination of one excised mass. Euthanasia of the animal was performed 11 days postsurgery and a proliferative lesion in the paralumbar musculature and similar, smaller proliferative lesions surrounding the right popliteal lymph node and in the ventricular wall of the heart were found. Metastatic lesions were found in the liver and lung. The histological features of the neoplastic tissues supported the diagnosis of a poorly differentiated, disseminated haemangiosarcoma. This is the first reported case of haemangiosarcoma in the Eastern barred bandicoot. PMID- 11022286 TI - An evaluation of a hand-held electrical resistance meter for the diagnosis of bovine subclinical mastitis in late lactation under Australian conditions. AB - OBJECTIVE: To assess the ability of a hand-held device to differentiate between infected and noninfected bovine mammary glands according to the electrical resistance of milk, under Australian conditions. DESIGN: A cross-sectional study. PROCEDURE: Milk samples were collected from 236 quarters of 60 cows selected from a commercial dairy herd with a high prevalence of mastitis. The true infection status of these quarters was determined using bacteriology. Various methods were used in an attempt to relate the electrical resistance of milk from each quarter to the presence or absence of infection in that quarter. RESULTS: Although the electrical resistance of milk from infected quarters was generally lower than that of noninfected quarters, the overlap of readings between the two populations limited the ability of this device to indicate accurately whether a quarter was infected. The use of methods comparing the readings from each of the four quarters of a single cow did not allow the reliable detection of infected cows. CONCLUSION: Although this device may have some practical advantages in comparison with some other methods of diagnosing subclinical mastitis, the predictive value of the method was generally poor. PMID- 11022287 TI - Digoxin therapy. PMID- 11022288 TI - Pregnancy in a cow with uterus didelphys. AB - Uterus didelphys is a congenital defect of monotocous species. This short contribution describes the condition in a cow that had a pregnancy in the right horn and a corpus luteum on the ipsilateral ovary. This finding demonstrates that pregnancy can occur in a cow with uterus didelphys, although the fate of the pregnancy was obviously not known. The case was identified during routine collection of reproductive tracts in a New Zealand abattoir. It is believed to be the first report of uterus didelphys in a bovine abattoir specimen. PMID- 11022289 TI - The case for solar light radiation being more significant than ambient temperature in producing lethal hyperthermic ergotism in cattle. PMID- 11022290 TI - Use of the polymerase chain reaction assay for the detection of Mycobacterium avium subspecies paratuberculosis in blood and liver biopsies from experimentally infected sheep. AB - OBJECTIVE: To evaluate a polymerase chain reaction-based assay for the detection of Mycobacterium avium subsp paratuberculosis in blood and liver biopsies from subclinically infected sheep. STUDY DESIGN: A direct PCR assay for the detection of M a paratuberculosis was applied to liver biopsy specimens and to samples of blood that were sequentially collected over 53 weeks from 14 sheep infected experimentally with the organism. RESULTS: Of 117 blood samples from the 14 experimentally infected sheep, two tested positive in the PCR assay. Both positive results were obtained in two subclinically infected sheep that had paratuberculosis later confirmed by histological examination at necropsy. However, the assay failed to detect the target DNA in samples of blood from five other sheep with histologically confirmed paratuberculosis. Similarly, the PCR assay on liver biopsy specimens collected 32 weeks after administration of M a paratuberculosis gave only two positive results, both of which were obtained in sheep with histological evidence of paratuberculosis. CONCLUSIONS: The PCR assay on blood and liver biopsies does not provide a useful method for the diagnosis of M a paratuberculosis infection in subclinically infected sheep. PMID- 11022291 TI - Disease and injury among veterinarians. AB - OBJECTIVE: To identify occupational causes of disease and injury in veterinarians and their staff. DESIGN: A self-administered questionnaire was mailed to all of the 160 registered veterinary practices in Western Australia. RESULTS: Injuries accounted for most workers' compensation claims over a 12-month period with 31% of respondents losing a total of 360 work days with a mean of 13.3 days whereas non-occupational diseases such as influenza accounted for 408 work days lost. Over a 10-year period, 71% of respondents had been injured. The major physical injuries were dog and cat bites, cat scratches, scalpel blade cuts and back injuries from lifting heavy animals. Exposure to chemicals such as flea rinses, formalin, glutaraldehyde, x-ray developers and gaseous anaesthetics were reported to cause headache, nausea and allergies. Thirty percent of respondents did not have extractor fans for scavenging waste anaesthetic gases. The study showed high use of radiography (94%) for diagnostic purposes with 24% of respondents believing radiation exposure is a major occupational health and safety issue. Stress, drug abuse, suicide and burglary were also reported. Despite a high awareness of zoonotic diseases, there were very few reports of these. CONCLUSION: Injuries and other occupational hazards reported together with work days lost demonstrate a need for improving the working environment of veterinarians and their staff and the development of comprehensive health and safety programs. PMID- 11022292 TI - Ribotype diversity of porcine Pasteurella multocida from Australia. AB - OBJECTIVE: To use the technique of ribotyping to investigate the genetic diversity of Australian isolates of Pasteurella multocida associated with outbreaks of clinical disease in Australian pigs. DESIGN: One hundred and seven porcine P multocida isolates were analysed by ribotyping using the restriction enzymes HpaII and HindIII. The genetic population structure of the Australian porcine P multocida isolates was determined through statistical analysis of the joint ribotype patterns, and this was then compared with biochemical and epidemiological data available for the population. RESULTS: A total of 25 combined ribotypes were recognised, which were grouped into five ribotype clusters. Despite the deliberate selection of diverse isolates, the study revealed only a limited degree of genetic diversity. Fourteen of the ribotypes contained multiple isolates, and 12 of these ribotypes were present on more than one farm. Three of the seven biovars analysed in the study showed very limited diversity. All fifteen biovar 2 isolates (subsp multocida) were found in a single cluster (III), while all four biovar 8 isolates, which correspond to P multocida subsp gallicida, were allocated by themselves to a single cluster (IV). All nine of the biovar 12 isolates (lactose-positive subsp multocida) were assigned to a single cluster (I), together with the single biovar 14 isolate, which was the only other lactose-positive isolate in the population (ODC-negative). CONCLUSION: A limited number of ribotypes of P multocida are associated with Australian pigs. The majority of these ribotypes are widely distributed across multiple farms, and across multiple states. Individual farms can possess multiple ribotypes of P multocida. Some of the unusual biochemical variants of P multocida present in Australian pigs have a very limited genetic diversity. The nature of pig production in Australia, primarily involving continuous flow systems with few closed herds, has possibly contributed to the widespread distribution of a limited number ribotypes. PMID- 11022293 TI - The isolation of enterohaemorrhagic Escherichia coli O111:H- from Australian cattle. PMID- 11022294 TI - Full-length infectious clone of a pathogenic Australian isolate of chicken anaemia virus. PMID- 11022295 TI - An unusual haemolymphoid mass in the retroperitoneum of a dairy cow. PMID- 11022296 TI - [Effect of hyperbaric air on biorhythms of the pain sensitivity and analgesic effect of morphine]. AB - The circadian and high-frequency (2-2.5 h) pain sensitivity rhythms were observed in male mongrel rats under natural illumination conditions (LD 24, January February). A minimum pain sensitivity threshold with respect to subcutaneous electrode stimulation was observed in the dark phase of the LD 24 cycle. Morphine injections (5 mg/kg, i.p.) led to inversion of the circadian rhythm and expansion of the high-frequency spectrum. The rhythm synchronism was broken under the hyperbaria conditions, whereby the circadian acrophase shifted to exhibit a maximum within the day light phase. The concurrent morphine administration on the hyperbaria background induced the back inversion of the circadian pain sensitivity rhythm. The high-frequency rhythm was not significantly affected by increased pressure--neither alone nor in combination with morphine injections. PMID- 11022297 TI - [Effects of opioid analgesics on potential-gated ion channels in the pond snail neurons]. AB - The effects of opioid analgesics morphine, promedol, tramadol, and butorphanol on the transmembrane calcium, sodium, and potassium (fast and slow) ion currents in pond snail (Lymnaea stagnalis) neurons were studied by intracellular dialysis and membrane potential (voltage-clamp) measurements. It was established that all these drugs produce a dose-dependent reversible inhibition of the ion currents (sodium calcium potassium) and reduce nonspecific leak currents, thus producing a stabilizing action upon the sample membrane. The current-inhibiting effect of morphine was not eliminated by naloxone (500 microM); moreover, the latter drug significantly inhibited the ion currents as well. PMID- 11022298 TI - [Effect of biotransformation on the receptor selectivity of muscarinic antagonists in vivo]. AB - In the experiments in vivo it is found that the receptor selectivity of muscarine antagonist glypine is time-dependent unlike atropine, amedine, benzhexol, benactyzine, and thropacin. Using modulation of the metabolic system activity it is shown that upon biotransformation glypine forms active metabolites that differs in receptor selectivity of the action. PMID- 11022299 TI - [Drug stimulation of the peripheral nerve regeneration]. AB - A comparative study of the effects of xymedone, riboxin, and piracetam upon the regeneration of myelinated axons and the number of surviving sensory neurons in spinal ganglia L4-L5 was studied on the model of sciatic nerve transection in rats. The three drugs decrease the number of neurons entering the posttraumatic apoptosis. Riboxin and xymedone stimulate the regeneration of myelinated axons, the latter drug being more effective and producing a 21.3 and 14.7% increase in the number of regenerated myelinated axons and a 29.3 and 37.7% increase in the relative number of sensory neurons (the transected/contralateral side ratio) in spinal ganglia L4-L5 relative to control by the 60th and 90th day upon transection, respectively. PMID- 11022300 TI - [Effect of beditine on the cerebral circulation and various metabolic parameters]. AB - Beditin, or 2-(2-amino-4-thiazolyl)-1,4-benzodioxane hydrochloride, is a new benzodioxane derivative possessing antihypoxant and alpha 2-adrenoreceptor blocker properties. In contrast to idazoxan, a well-known alpha 2-adrenoreceptor blocker, beditin (3 mg/kg) significantly increases the circulation rate and decreases the blood vessel resistance in various brain structures of rats and cats without considerably changing their systemic hemodynamic indices. Upon a one side ligation of the common carotid artery in rats, beditin prevented a sharp increase in the lactate dehydrogenase activity and a growth in the content of free ammonia in the brain, which are characteristic of the ischemic states. The ammonia-eliminating effect of beditin is related to an increase in the glutamate dehydrogenase activity in the reductive amination reaction. PMID- 11022301 TI - [Pharmacological analysis of the role of central alpha-2 adrenergic and imidazoline receptors in mechanism of the hypotensive effect of clonidine in rats]. AB - The role of alpha 2-adrenergic and imidazoline receptors in the hemodynamic response to clonidine (manifested by reduced arterial pressure, heart rate, and renal blood flow rate) was studied by injecting their antagonists with different affinity (yohimbine and idazoxan) into narcotized Sprague--Dawley rats. The local introduction (microinjection into rostral ventrolateral medulla) and systemic administration (intraperitoneal injection) of the drugs showed that the hemodynamic effect of clonidine is mediated predominantly by the central alpha 2 adrenergic receptors, rather than by the imidazoline receptors. PMID- 11022302 TI - [Anti-arrhythmic effect of phytoadaptogens]. AB - Repeated prophylactic administration of plant adaptogen preparations based on extracts from rhodiola, eleutherococcus, leuzea, and ginseng, produced a pronounced antiarrhythmic effect on the model of adrenal arrhythmia in animals. Preliminary opioid receptor block by naloxone reduced the protective effect of phytopreparations in the adapted animals. Intravenous and intracerebroventricular administration of naloxone (opioid receptor antagonist) showed that the anti arrhythmic effect of rhodiola extract proceeds through the activation of both central and peripheral opioid receptors. PMID- 11022303 TI - [Effect of instenon on the uterine contractile activity]. AB - The effect of instenon upon the uterine contractile activity was studied in pregnant rats. The drug administration at a dose resulted in significant suppression of the myometrium contractility. The effect was not decreased when instenon was injected against the background of oxytocin. On the other hand, the concurrent injection of oxytocin upon a single or long-term administration of instenon produced a pronounced stimulating action on the myometrium contractility. PMID- 11022304 TI - [Effect of ecdystene on parameters of the sexual function under experimental and clinical conditions]. AB - The effects of ecdysterone and the related drug ecdysten on the sexual activity were studied under experimental and clinical conditions. A 10-day administration of ecdysterone (5 and 10 mg/kg, p.o.) improved behavioral characteristics of the sexual function in rats, the effect being especially pronounced during the fist days of experiment. The administration of ecdysten to men with the infertility diagnosis (disturbed spermatogenesis as a complication of some urologic diseases) increased the copulative function and improved the sperm quality. The administration of ecdysten to patients in the stage of recovery upon myocardial infarction also improved the sexual function. PMID- 11022305 TI - [Characteristics of lipid peroxidation in various tissues during acute stress and its correction by pyracetam and cerebrolysin]. AB - The process of lipid peroxidation (LPO), the activity of antioxidant enzymes, and the effects of piracetam (300 mg/kg) and cerebrolysin (0.1 ml/kg) were studied in the brain, liver, thymus, spleen, and blood serum of white male rats under acute stress (immobilization) conditions. The results of analyses confirmed that the stress syndrome development involves the LPO acceleration and retardation stages not coinciding in various tissues. The antioxidant effect of the nootropic agents in CNS has a permanent character, being independent of the initial status of the animal organism. The effect of both drugs upon the LPO level in peripheral tissues has a modulative character and can be considered as a sum of the central antistressor action and the direct influence upon the functional activity of effectors. PMID- 11022306 TI - [The mineral dietary composition affects glucose metabolism and modulates insulin effect in intact rats]. AB - The experiments on intact rats showed that an excess sodium chloride consumption and the use of giposol (NaCl substitute) produce a hypoglycemic effect. Giposol increases, whereas NaCl decreases, the glucose tolerance. Giposol activates hexokinase and glucose-6-phosphate dehydrogenase (the key enzymes of the glucose metabolism in tissues) and reduces the effect of insulin. Both giposol and NaCl increase the level of glucose absorption in small intestine. The gastric motility is activated by NaCl and not affected by giposol. PMID- 11022307 TI - [Immunopharmacologic properties of 12-iminosubstituted 8-aza-D-homogonan and trimezon]. AB - The immunopharmacological properties of two new compounds with an 8-azasteroidal structure, representing the salts (hydrochloride and acetate) of 12-imino substituted 8-aza-D-homogonan, were studied in comparison with the properties of the immunostimulant trimeson. Similar to trimeson, both new compounds produce a dose-dependent activation of the humoral chain of immunogenesis in the model of primary immune response to goat erythrocytes in CBA, C57BL/6, and C57W mice. The immunostimulant activity of the 8-aza-D-homogonan derivatives is inversely proportional to the antigen load (2 x 10(7)-2 x 10(9) goat erythrocytes per mice) and depends to a considerable extent of the genotype of experimental animals. A modification of the 8-aza-D-homogonan structure by substituting imino group for oxygen at position 12 of the 8-azasteroidal skeleton leads to a sharp increase in toxicity as compared to that of trimeson. PMID- 11022309 TI - [Evaluation of major pharmacokinetic parameters of high molecular weight polyethylene oxide using polymer hydrodynamic properties]. AB - The pharmacokinetics of high-molecular-weight polyethylene oxide (HMW PEO)--a parent compound in the polietoks drug--was experimentally studied in cats. A PEO concentration in the blood was determined by measuring a decrease in the hydrodynamic resistance for a blood sample flowing in the turbulent mode via a glass capillary. Variation of the polymer concentration in the blood is described by a single-exponent function with an elimination halftime of T1/2 = 3.15 h. According to the calculation, 99.6% of the initial amount of PEO losses the antiturbulent activity within 24 h. PMID- 11022308 TI - [Modification by lacidipine of the clastogenic effect of dioxidine in vivo]. AB - The influence of lacidipine (0.1-10 mg/kg, intragastric) on the clastogenic effect of dioxidine (100 and 200 mg/kg, i.p.) under conditions of their single and repeated (five-fold, 24-h interval) administration was studied by the chromosome aberration assay in the metaphase bone marrow cells of BALB/c and C57BL/6 male mice. It was found that single (5 or 10 mg/kg) and repeated (10 mg/kg) introduction of lacidipine enhances the clastogenic effect of dioxidine in both genotypes. At the same time, a single treatment of C57BL/6 mice with 0.1 and 1 mg/kg of lacidipine sometimes significantly reduced the clastogenic effect of dioxidine (200 mg/kg). Thus, lacidipine exhibits a comutagen effect in vivo when administered at large doses (5 and 10 mg/kg) but not at small doses, where the drug sometimes acted as antimutagen in C57BL/6 mice. PMID- 11022310 TI - [Toxic effect of the novel phenol antioxidant SO-3 in the subchronic experiment]. AB - Subchronic tests on rats showed that a new phenolic antioxidant bis[3,(3, 5-di tert-butyl-4-hydroxyphenyl)propyl] sulfide (SO-3) exhibits low toxicity upon peroral administration. Neither significant variations in the hematological and integral indices, nor pathological shifts in the activity of serum enzymes or pathomorphological changes in the internal organs were observed. Simultaneous administration of a sunflower oil with SO-3 led to a significant decrease in the blood cholesterol level, produced a hepatoprotector effect, and exhibited a protective action with respect to the mucous membranes of the gastrointestinal tract. Animals receiving large doses of SO-3 showed no signs of a prelipid stage of the arteriosclerosis typically observed in the control group. PMID- 11022311 TI - [Preclinical toxicological study of ecorsin]. AB - A new drug preparation ecorsin based on the dry aspen bark extract was studied for toxicological safety on a preclinical stage. The drug exhibited no toxicity upon a single administration to rats and mice (both male and female). The intragastric administration of ecorsin for 3 months to rats (at a daily dose of 50, 250, and 500 mg/kg) and rabbits (25 and 50 mg/kg) led to neither functional not morphological changes in hemopoietic and lymphoid organs, liver, kidney, heart, digestive system, and CNS. The long-term administration resulted in a partial atrophic modification of convoluted seminiferous tubules in impuberal male rats, while not affecting the testes of aged animals. The administration of ecorsin at 50, 250, and 500 mg/kg led to dose-dependent changes in some characteristics of the reproductive capacity in rats. Ecorsin did not modify the extent of allergic reactions and produced no immunotoxicant and mutagen effects. PMID- 11022312 TI - [Histamine blockers increase efficacy of specific antidote prophylaxis of intoxication by cholinesterase inhibitors in mice]. AB - Dimedrol and pipolphen, but not loratadine, increase efficacy of the atropine and dipiroxime treatment of mice--an antidote prophylaxis against their intoxication with phosphacol and aminostigmine. The results of experiments confirm a hypothesis that histamine participates in the formation of secondary toxicity reactions in the case of heavy poisoning with anticholinesterase agents. PMID- 11022313 TI - [Effect of pyrimidine derivatives on the skin reparative regeneration during stress in laboratory animals]. AB - Experiments on rats showed that some pyrimidine derivatives stimulate the skin repair in animals with thermal and chemical burns under stress conditions. The efficacy of compounds tested increases in the following order: 2-methyl-4-amino-6 hydroxypyrimidine < hydroxymethyluracil < methyluracil. PMID- 11022314 TI - [Correction by common sea buckthorn bark and sprout extracts of hormonal and metabolic disturbances during stress in rats]. AB - Long-term administration of the common sea buckthorn bark and sprout extract improves the hormonal-metabolic organism status in rats disturbed by a stress factor (immobilization). The drug administration led to normalization of the altered functional activity indices of the neuro-endocrine system (disturbed adrenocorticotropin, 11-deoxycortisol, insulin, urea, and glucose levels) by affecting the production of glucocorticoids and increasing the hypothalamus sensitivity with respect to regulatory signals. PMID- 11022315 TI - [Effect of angiotensin II and antagonists of the renin-angiotensin system on the cerebral circulation]. PMID- 11022316 TI - Defibrillator/monitor/pacemakers. AB - Defibrillator/monitors allow operators to assess and monitor a patient's ECG and, when necessary, deliver a defibrillating shock to the heart. When integral noninvasive pacing capability is added, the resulting device is referred to as a defibrillator/monitor/pacemaker. In this Update Evaluation, we present our findings for nine such units, including complete Product Profiles for two newly evaluated models and update information for seven other models evaluated in our May-June 1993 and February 1998 studies. We tested the two newly evaluated models using the same basic protocol as in our previous studies. However, we did add some new tests--and revise some old ones--to account for advances in the technology. These advancements include the increasing use of advisory modes and the increasing availability of expanded monitoring capabilities (which allow units to function, at least to some degree, like a physiologic monitor). As in our previous studies, we rated each model separately for three common defibrillation applications: (1) general crashcart use, (2) in-hospital transport use, and (3) prehospital use by emergency medical services (EMS) personnel. Because each application requires its own set of capabilities, it's not surprising that few models are appropriate for all applications. However, we did identify three models that perform well--earning a rating of either Acceptable or Preferred--in all three areas. PMID- 11022317 TI - New frequencies for medical telemetry. FCC's plan is final--now what? AB - The frequency bands used by wireless medical telemetry in the United States are getting crowded. More and more competing users are occupying those frequencies, putting medical telemetry at an ever-greater risk for harmful interference. In our May 2000 Evaluation of physiologic monitoring systems, we discussed the U.S. Federal Communications Commission's (FCC) proposal to allocate new frequency bands in which medical telemetry could operate in the United States as a primary user. (Being a primary user means that medical telemetry is protected against interference from other sources in the specified frequency bands.) On June 12, 2000, FCC released a Report and Order (FCC No. 00-211) making new frequencies available for medical telemetry use on a primary basis. These new frequencies, called the Wireless Medical Telemetry Service, will be administered by a frequency coordinator and should help alleviate the crowding of available frequencies. The final rules presented in the Report and Order will go into effect on October 16, 2000. In this article, we discuss why these new frequencies are needed and what you should do about them. PMID- 11022318 TI - Unnoticed alarms contribute to a death in an MRI suite. PMID- 11022320 TI - Aiming beam degradation in Laserscope Pulsar/Paragon CO2 laser. PMID- 11022319 TI - Staff injuries caused by falling siderail on bariatric rehabilitation platform bed supplied by Wheelchairs of Kansas. PMID- 11022321 TI - Deuterium labelled nonylphenols in an in-vitro model of percutaneous absorption of environmental xenoestrogens. AB - A technical mixture of isomeric nonylphenols NP is formed as rather persistent degradation product of nonionic surfactants and has become widespread in the environment, e.g., in surface waters. Information about possible pathways for incorporation is needed for risk assessments, as NP has estrogenic properties. NP uptake after skin contact was determined using isolated and perfused bovine udders as models for human skin. NP-d2 labelled in the positions ortho to the OH group was prepared by an exchange procedure which did not measurably change the relative amount of isomers. Samples of udder perfusate, milk equivalent produced during the experiments and skin tissue were taken immediately before administration of 500 mg or 50 mg NP-d2 on an udder skin area of 200 cm2 and then for 5 h. They were analyzed by GC-MS for NP and NP-d2 after cleanup by steam distillation and final extraction with SPME fibres. The results are an unambiguous proof of the penetration of NP into the capillary system of living mammals after skin contact. PMID- 11022322 TI - Experience with the [13C2]oxalate absorption test. AB - Hyperoxaluria is the most important risk factor for a formation of calcium oxalate-urinary stones. Usually, the bulk of oxalate will be formed in the human body, but in many patients the oxalate from food plays the decisive role. Conventionally, in urine the endogenous oxalate can not be distinguished from food derived oxalate. We have developed a standardized oxalate-absorption test, applying a physiological dose (50 mg disodium salt of [13C2]oxalic acid) of labelled oxalate. The assay has been published. Now we report on the first extensive applications of this test in 86 volunteers and 135 patients from different groups with calcium oxalate stones or an increased risk of the formation of such stones. In one-third of the patients with calcium oxalate urinary stones an oxalate hyperabsorption was diagnosed. For these patients, a dietetic stone prophylaxis and/or therapy is indicated. PMID- 11022323 TI - Transfer of symbiotically fixed nitrogen in an alfalfa-grass mixture studied through isotope dilution in a pot experiment. AB - The nitrogen transfer between alfalfa and ryegrass was studied through isotope dilution at three different levels of N fertilization (20 mg N/pot, 200 mg N/pot, 400 mg N/pot) in a pot experiment using quartz sand as a substrate. An isogenic, nodulating, but non nitrogen fixing alfalfa line was used as a reference crop. Fixed N was transferred to the grass in the 20 mg N treatment and contributed markedly to the N nutrition of the grass (about 50% of the N in the plants). No transfer of fixed N could be detected in the higher fertilized treatments, although nitrogen fixation was only slightly inhibited by the presence of the fertilized mineral N. It is concluded that N transfer is strongly influenced by the N concentration in the substrate and transferred N contributes only slightly to the productivity of the legume/grass mixture under the given experimental conditions. PMID- 11022324 TI - 13C and 15N distributions in three spodic dystric cambisols under beech and spruce. AB - The study of natural isotopic abundance signatures is useful to gain further insights in the processes resulting in depthwise changes in the composition of soil organic matter (SOM). Objectives were to describe the delta 13C and delta 15N abundances of SOM with depth in soils from a 153-year old beech (B1), a 119 year old spruce (F1) and a 61-year old spruce (F2) stand at Solling, north-west Germany, and to study, how podzolisation affects the isotopic abundances of 13C and 15N in the SOM. The degree of podzolisation decreased in the order F1 > B1 > F2. At the surface of the humus layer of all three sites, delta 13C values are approximately 1 to 4/1000 higher than in the leaves and needles, probably mainly due to the discrimination of 13C by microbial decomposition. 13C abundances in the organic layers of B1 and F2 increased only slightly from -27.6/1000 PDB (B1, L) to -27.2/1000 PDB (B1, Oh) and from -26.3/1000 PDB (F2, L) to -25.9/1000 PDB (F2, Oh), suggesting that biotic activity resulted in mixing of organic matter. At F1, however, 13C abundance increased from -27.5/1000 PDB (L) to -26.0/1000 PDB (Oh) which reflects the lack of mixing by animals. In the upper 2-4 cm of the mineral soil, i.e., in the eluvial horizons Aeh, 13C values showed a minimum at the spruce sites which was presumably related to a translocation of 13C enriched fulvic acids. Depthwise changes in delta 15N values were not related to podzolisation processes. At all three sites, a 13N enrichment with depth occurred in the mineral soil which is the result of the discrimination of 15N by microbial decomposition. PMID- 11022325 TI - Use of 13C and 15N mass spectrometry to study the decomposition of Calamagrostis epigeios in soil column experiments with and without ash additions. AB - The dynamics of C and N in terrestrial ecosystems are not completely understood and the use of stable isotopes may be useful to gain further insight in the pathways of CO2 emissions and leaching of dissolved organic carbon (DOC) and nitrogen (DON) during decomposition of litter. Objectives were (i) to study the decomposition dynamics of Calamagrostis epigeios, a common grass species in forests, using 13C-depleted and 15N-enriched plants and (ii) to quantify the effect wood ash addition on the decomposition and leaching of DOC and DON. Decomposition was studied for 128 days under aerobic conditions at 8 degrees C and moisture close to field capacity in a spodic dystric Cambisol with mor-moder layer. Variants included control plots and additions of (i) Calamagrostis litter and (ii) Calamagrostis litter plus 4 kg ash m-2. (i) Decomposition of Calamagrostis resulted in a CO2 production of 76.2 g CO2-C m-2 (10% of added C) after 128 days and cumulative DOC production was 14.0 g C m-2 out of which 0.9 g C m-2 was Calamagrostis-derived (0.1% of added C). The specific CO2 formation and specific DOC production from Calamagrostis were 6 times higher (CO2) and 4 times smaller (DOC) than those from the organic layer. The amount of Calamagrostis derived total N (NH4+, NO3-, DON) leached was 0.7 g N m-2 (4.8% of added N). Cumulative DON production was 0.8 g N m-2 which was slightly higher than for the control. During soil passage, much of the DOC and DON was removed due to sorption or decomposition. DOC and DON releases from the mineral soil (17 cm depth) were 6.3 g C m-2 and 0.5 g N m-2. (ii) Addition of ash resulted in a complete fixing of CO2 for 40 days due to carbonatisation. Afterwards, the CO2 production rates were similar to the variant without ash addition. Production of DOC (98.6 g C m 2) and DON (2.5 g N m-2) was marked, mainly owing to humus decay. However, Calamagrostis-derived DOC and Calamagrostis-derived total N were only 3.9 g C m-2 (0.5% of added C) and 0.5 g N m-2 (3.4% of added N). The specific DOC production rate from the organic layer was 6 times higher than that from Calamagrostis. The results suggest that with increasing humification from fresh plant residues to more decomposed material (OF and OH layers) the production ratio of DOC/CO2-C increases. Addition of alkaline substances to the forest floor can lead to a manifold increase in DOC production. PMID- 11022326 TI - A dual 13C and 15N long-term labelling technique to investigate uptake and translocation of C and N in beech (Fagus sylvatica L.). AB - A continuous dual 13CO2 and 15NH4(15)NO3 labelling experimental set-up is presented that was used to investigate the C and N uptake and allocation within 3 year old beech (Fagus sylvatica L.) during one growing season. The C and N allocation pattern was determined after six, twelve and eighteen weeks of growth. The carbon uptake was distinctly different in the three phases examined: The first six weeks after budbreak were dedicated to leaf growth with a R/S (root to shoot) ratio of 0.14 for the new carbon. The second growth phase showed a balanced R/S ratio of C allocation and after week 13, the root compartment was the main carbon sink (R/S = 6.97). Nitrogen allocation was more basipetal as compared to carbon. In the second growth phase, R/S of Nnew was 5.57 but fell to 3.54 for the third growth phase probably due to formation of reserves in buds and stem. PMID- 11022327 TI - Net and gross nitrogen mineralization in drained and reflooded fen soils. AB - One purpose of new land use concepts for degraded fens (organic soils with high N content) is the reduction of the mineralization process due to very high groundwater levels. However, knowledge of nitrogen mineralization process (net and gross) in degraded fen soils affected by reflooding is very small. Therefore, the objectives of our study were (a) to evaluate the suitability of 15N pool dilution method for measurements of gross mineralization rates in degraded fen soils and (b) to investigate how the reflooding of a degraded fen affects the net and gross nitrogen mineralization in a short-term incubation experiment. The usability of the 15N pool dilution method was diminished by the low recovery of the applied 15NH4+ at time zero. The recovery of the added 15NH4+ in the extractable soil NH4+ pool was only 13.5% for the drained soil and 59.6% for the reflooded soil. However, the gross mineralization rates were similar for both soils and exceeded always the net rates substantially. The cumulative net mineralization rate was higher for the reflooded soil (1.58 micrograms N*cm-3*d 1) than for the drained soil (-0.67 microgram N*cm-3*d-1). Differences between the two soils were also found in the nitrification intensity and the loss of 15N. This was probably one reason for the higher net mineralization rate in the reflooded soil. PMID- 11022328 TI - Evidence-based practice: ANAC taking the lead. PMID- 11022329 TI - Women with HIV infection: the three waves of scientific inquiry. AB - Initial efforts to curtail the spread of HIV infection to women failed. Consequently, women are the fastest growing population newly infected with HIV. This article presents a historical look at the waves of scientific inquiry that directed research on HIV infection among women. The author proposes three distinct waves. Each has been shaped by movement within the scientific community away from traditional biomedical and public health approaches toward feminist strategies that embrace the social, political, and cultural forces that influence women's health. These waves provide both valuable insights for nurses new to the field of HIV and a framework to guide future research on women with HIV infection. PMID- 11022330 TI - Empowerment: a case study of a grandmother caring for her HIV-positive grandchild. AB - A case study methodology is used to explore the meaning and development of empowerment in a grandmother who is the primary caregiver of a child with HIV infection. Empowerment to this grandmother meant being strong, coming to grips with life's challenges, and then moving on. Empowerment developed over the grandmother's life span as a result of developmental and situational experiences such as motherhood, disclosure of the HIV diagnosis, sharing her positive and negative feelings with others, gaining knowledge about HIV, making management decisions, and her increasing sense of spirituality. This grandmother manifests her empowerment through her public commitment to influence others positively about HIV infections through speaking, writing, and leadership in groups. Clinical implications are discussed. PMID- 11022331 TI - Complementary and alternative therapies to manage HIV-related symptoms. AB - Persons with HIV infection report substantial use of complementary and alternative medical (CAM) therapies for symptom management. Anecdotal reports from patients indicate that CAM approaches are helpful; however, there is limited scientific information on the safety and efficacy of these therapies in the HIV population. The purpose of this review is to critically appraise the scientific evidence for selected CAM therapies that are used by HIV-infected persons to manage three common symptoms: nutritional alterations, pain, and depression. PMID- 11022332 TI - Correcting deficiencies in HIV/AIDS care for transgendered individuals. AB - Studies suggest that transgendered individuals are at high risk for acquiring HIV through injection drug use and sexual risk behaviors. Beginning efforts are being directed to identify the unique needs and concerns of these individuals and to develop culturally sensitive programs that will be successful in recruiting and retaining these individuals in drug abuse and HIV treatment services. These efforts include examining the available and needed services from the perspectives of both the transgendered community and health care professionals. This article reviews the phenomenon of transgenderism and the social context that places these individuals at risk for HIV. Additionally, this article proposes some guidelines for those who are in positions responsible for designing, evaluating, and implementing existing health care services to transgendered populations. PMID- 11022333 TI - Sexual prevention of HIV: a model for Latinos. AB - Despite reductions in AIDS deaths in the general population, Latino AIDS deaths in the United States have been increasing. The underlying cause of the rising HIV infection rates in the U.S. Latino population is not fully understood. Focus groups among Mexicans in North Carolina revealed methods of prevention that were different from those recommended by the Centers for Disease Control and Prevention (CDC). A research-based Mexican model of HIV prevention is proposed and contrasted with a model deducted from CDC prevention strategies. Recently immigrated Mexicans may be unlikely to adopt the CDC model of HIV prevention due to their culturally specific views of prevention. PMID- 11022334 TI - Knowledge, self-efficacy, coping, and condom use among urban women. AB - In this study, path analysis was used to test a model of the relationships among condom use knowledge, self-efficacy for condom use, coping, and condom use in a sample of 100 urban women. In the final model, the paths between condom use knowledge and self-efficacy, between self-efficacy and condom use, and between self-efficacy and problem-focused coping were significant. In addition, condom use knowledge was indirectly related to condom use through self-efficacy. The final model accounted for 43% of the variance in condom use. Although the women engaged in risky behaviors including having multiple partners, high-risk sexual partners, and unprotected intercourse, they did not worry about or perceive themselves at risk for HIV. To assist women to decrease their risk for sexually transmitted HIV infection, clinicians should conduct individualized assessments of each client's sexual behaviors to identify women at risk. Individualized interventions are recommended to increase both knowledge of how to use condoms and self-efficacy for condom use among women at risk for HIV. PMID- 11022335 TI - Lactic acidosis associated with nucleoside reverse transcriptase inhibitors. AB - Limitations on antiretroviral therapies as a result of resistance and adverse events have been described in the literature. Lactic acidosis has been reported in patients on nucleoside reverse transcriptase inhibitors on rare occasions. Successful assessment and treatment of lactic acidosis is dependent on the nurse's role in the recognition of this condition. This article provides an overview of the clinical presentation, diagnosis, treatment strategies and nursing management of lactic acidosis in patients with HIV/AIDS. PMID- 11022336 TI - Program evaluation, revisited: a community-based HIV prevention grant. PMID- 11022337 TI - Ruminant adaptation to negative energy balance. Influences on the etiology of ketosis and fatty liver. AB - Ketosis and fatty liver occur when physiologic mechanisms for the adaptation to negative energy balance fail. Failure of hepatic gluconeogenesis to supply adequate glucose for lactation and body needs may be one cause of ketosis; however, poor feedback control of nonesterified fatty acid release from adipose tissue is another likely cause of ketosis and fatty liver. The types of ketosis resulting from these two metabolic lesions may require different therapeutic and prophylactic approaches. PMID- 11022338 TI - Subclinical ketosis in lactating dairy cattle. AB - Subclinical ketosis is an important and common condition of early-lactation dairy cattle. It is associated with losses in milk production and increased risk of periparturient disease. Prevention depends on several factors, including proper transition-cow nutrition, management of body condition, and the use of certain feed additives such as niacin, propylene glycol, and ionophores. All currently available cowside tests for subclinical ketosis have certain limitations in their use. Effective monitoring schemes for subclinical ketosis can be developed, however, and these may be useful in many herd health programs. PMID- 11022339 TI - Metabolic aspects in the etiology of displaced abomasum. AB - Results of studies on metabolic variables that exist prior to the diagnosis of displaced abomasum (DA) indicate that subclinical ketosis is a risk factor for DA. Serum aspartate aminotransferase activity, serum and milk betahydroxybutyric acid concentrations, and milk fat-to-protein ratios were often elevated before the diagnosis of DA, and these variables may be used to predict DA. Prevention of subclinical ketosis may prevent DA. PMID- 11022340 TI - Glucagon as a potential therapy for ketosis and fatty liver. AB - An experiment demonstrating the usefulness of glucagon as a treatment for fatty liver and ketosis in early-lactation dairy cows has been described. This is the first report of the ability of glucagon, or any agent, to promote clearance of lipid from livers of animals suffering hepatic lipidosis. The development and use of glucagon as a therapeutic agent for the fatty-liver complex in dairy cows may provide a powerful management tool to enhance the profitability of the high producing dairy cow. PMID- 11022341 TI - Dry cow management for the prevention of ketosis and fatty liver in dairy cows. AB - Dramatic increases in energy requirements during late gestation and early lactation, superimposed on an animal with a profound drop in DMI just before calving, make the dairy cow highly susceptible to the metabolic diseases ketosis and hepatic lipidosis. Increased serum concentrations of NEFA appear to be causally linked to these problems, and feeding strategies to reduce or avoid this dramatic increase are desirable for optimal health and performance. During the last 3 to 4 weeks prepartum, a diet higher in energy and protein concentration than current NRC recommendations should be fed so that adequate nutrient intake occurs within the limits of the reduced DMI. The additional energy should be provided by glucose precursors, such as starchy concentrates or propylene glycol, and not by lipid. Excessive energy and reduced fiber should be avoided both early in the dry period (more than 28 days prepartum) and immediately postpartum. Attention should be paid to the environment of the cow, especially during the last 3 weeks prepartum, to avoid environmental stressors as much as possible. PMID- 11022342 TI - Pregnancy toxemia of ewes, does, and beef cows. AB - Pregnancy toxemia commonly affects pregnant ewes and does during late gestation. This metabolic disease is thought to result from disruption of the dam's glucose homeostatic mechanism in response to increased nutritional demands of the rapidly developing fetal placental unit. Commercial production systems are comprised of a variety of nutritional, metabolic, genetic, physiologic, environmental, economic, health, and management factors that singularly or as a group influence the clinical expression of pregnancy toxemia. Recognizing the role management plays in controlling these inputs is crucial to pregnancy toxemia prevention and treatment programs. PMID- 11022343 TI - Pathophysiology of calcium and phosphorus disorders. AB - Hypocalcemia and hypophosphatemia are relatively common in periparturient cows. This article reviews the etiologic factors that contribute to the development of these conditions. The physiologic role of magnesium in calcium homeostasis and the physiologic effect of dietary cation-anion difference at the cellular level are discussed in depth. A theory to explain the development of periparturient hypophosphatemia is developed. PMID- 11022345 TI - Management of dry cows for the prevention of milk fever and other mineral disorders. AB - Mineral disorders in dairy cattle typically occur soon after calving and are often specifically related to nutritional management during the late dry period. The incidence of hypocalcemia, hypophosphatemia, and hypomagnesemia can be minimized by proper diet formulation and good overall nutritional management in the dry period. The addition of anions to the diet during the late dry period may be of particular benefit for preventing hypocalcemia; however, supplementation with anions will reduce dry matter intake if feeding management is not excellent. PMID- 11022344 TI - Pathophysiology of grass tetany and other hypomagnesemias. Implications for clinical management. AB - Magnesium is an essential mineral with many physiologic and biochemical functions. Surprisingly, Mg homeostasis is not regulated by a hormonal feedback system, but simply depends on inflow (absorption) from the gastrointestinal tract and outflow (endogenous secretion, requirement for milk production, uptake by tissues). Any surplus (inflow greater than outflow) is excreted via urine. Conversely, if the outflow (mainly milk secretion and endogenous loss) exceeds inflow, hypomagnesemia occurs because of the lack of hormonal mechanisms of homeostasis. The major reason for insufficient inflow is a reduced absorption of Mg from the forestomachs. Recent studies from our laboratory and data from the literature permit the proposal of a putative transport model for the secondary active transport of Mg across the rumen epithelium. This model includes two uptake mechanisms across the luminal membrane (PD-dependent and PD-independent) and basolateral extrusion via a Na/Mg exchange. The well-known negative interaction between ruminal K concentration and Mg absorption can be explained on the basis of this model: an increase of ruminal K depolarizes the potential difference of the luminal membrane, PDa, and as the driving force for PD dependent (or K-sensitive) Mg uptake. Because Na deficiency causes an increase of K concentration in saliva and ruminal fluid, Na deficiency should be considered a potentially important risk factor. The data obtained from in vitro and in vivo studies on the association of Mg transport, changes of ruminal K concentration, and PDa are extensive and confirm the model, if the ruminal Mg concentrations are below 2 to 3 mM. It is further proposed by the model that the PD-independent Mg uptake mechanism is primarily working at high ruminal Mg concentration (above 2 mM). Mg absorption becomes more and more independent of ruminal K with increasing Mg concentration, which can be considered as an explanation for the well-known prophylaxis of hypomagnesemia by increasing oral Mg intake. Fermentation products, NH4+ and SCFA, influence Mg absorption. The possible meaning regarding the pathogenesis of hypomagnesemia is not quite clear. A sudden increase of ruminal NH4+ should be avoided, because high NH4+ concentrations transiently reduce Mg absorption. The most prominent signs of hypomagnesemia are excitations and muscle cramps, which are closely correlated with the Mg concentration in the CSF. It is suggested that the clinical signs are caused by spontaneous activation of neurons in the CNS at low Mg concentrations, which leads to tetany. Prophylactic measures are discussed in context with the known effects on ruminal Mg absorption. PMID- 11022346 TI - Variability characteristics and test selection in herd-level nutritional and metabolic profile testing. AB - Nutritional assessment based on animal response factors is the basis of essentially all dietary recommendations. Blood concentrations of nutrients, metabolites, and hormones are important animal response factors associated with nutriture, making blood analysis an important nutritional assessment technique. There are, however, numerous sources of variability, other than nutrition, affecting the concentration of blood analytes used in nutritional assessment. Minimizing the effects of non-nutrient sources of variability and maximizing the effects of nutritional variability is the objective in designing strategies for blood sampling and testing for nutritional assessment. Important non-nutrient sources of variability are age, sex, gestation stage, lactation stage and milk yield, and season. When interpreting test results, grouping animals by these characteristics is an important means of minimizing the effects of non nutritional variability. Within these groups, it is important to take an adequate number of samples, generally starting out with at least seven. Finally, selecting appropriate tests is critical. Tests commonly used for clinicopathologic evaluations are not necessarily the best tests for nutritional assessment. Analytes should be chosen that are likely to have a large portion of their total variability caused by nutritional effects. This generally does not include those metabolites the blood concentrations of which are rigidly controlled by homeostatic forces. PMID- 11022347 TI - [Medicine at the time of the discovery of Brazil and in the beginning of the year 2000]. PMID- 11022348 TI - [Total serum IgE level in atopic and non-atopic individuals in Porto Alegre]. AB - PURPOSE: We determined the total serum IgE levels among healthy individuals and atopic patients with respiratory symptoms in Porto Alegre. METHODS: Atopic patients (n: 92) and normal controls (n: 86) were stratified according to age and sex. Presence of atopy was recorded by history, clinical examination and skin prick test reactivity to common aeroallergens. Serum IgE was measured by Pharmacia Immuno-Cap System. RESULTS: It was demonstrated that total serum IgE is higher in males than in females. Total serum IgE levels were significantly higher in the atopic group (404 UI/mL) in comparison with the non-atopics controls (54.4 UI/mL). Our study provided the normal range for total serum IgE among atopics and non-atopics individuals in Porto Alegre. CONCLUSIONS: The profile of serum IgE among our population (atopics and non-atopics) is similar to the one observed in other countries. PMID- 11022349 TI - [Esophagogastric acute injury caused by a chemical agent]. AB - Treatment of chemical agent ingestion remain controversial. The incidence of these episodes has increased over the several last decades due to a variety reasons. PURPOSE: To analyze the occurrence, complications and results of the treatment of chemically induced esophagogastric injury. METHODS: Twenty-one adult patients with chemically induced esophagogastric injury were retrospectively studied. The patients were admitted up to 23 days after ingestion of a chemical agent to the Emergency Department of Santa Casa of Sao Paulo University Hospital from August, in a 12-year period. The mean age was 32.1 years. Eleven patients were of the female gender, which attempted suicide. Soda was the most ingested agent (76.2%), muriatic acid was present in three cases (14.3%) followed by one case of sulfur acid and another one of ammonia (4.8% each). RESULTS: Injuries of the Larynx and Pharynx were frequently associated with those of the esophagus, accounting for 18 cases (85.7%). Esophageal, gastric and duodenal injuries were assessed and classified according to endoscopic features. Five cases each of severe esophageal or gastric lesions were present. CONCLUSION: Treatment and outcome varied and suggested placement of esophageal tube to be harmful. Global mortality rate was 28.6% with the highest rate related to esophageal injuries of the third degree. PMID- 11022350 TI - [Evaluation of inflammatory activity in chronic osteomyelitis. Contribution of scintigraphy with polyclonal antibodies]. AB - BACKGROUND: Active chronic osteomyelitis or complicating osteomyelitis are difficult to be diagnosed by radiological imaging modalities, such as plain radiograph and CT. They frequently cause increased bone remodeling, leading to nonspecific uptake of Tc-99m-bone scan agents and gallium-67. New radiopharmaceuticals with greater infection avidity are being developed, including the nonspecific polyclonal immunoglobulin (IgG) labeled with technetium 99m. Tc-99m-IgG may be available as a ready to use kit, with no reported side effects, low patient absorbed radiation dose and low cost. MATERIAL AND METHODS: 23 bone segments with suspected active chronic osteomyelitis or violated bone osteomyelitis were studied by Tc-99m-IgG scintigraphy. All patients underwent standard three-phase bone scintigraphy using methylene diphosphonate (Tc-99m MDP), gallium-67 scintigraphy and plain radiographs, compared with clinical evaluation and laboratory tests values. RESULTS: Infection was found in 8 sites. Sensitivity and specificity for Tc-99m-MDP, gallium-67 and Tc-99m-IgG scintigraphy were, respectively, 88 and 36%, 75 and 73%, 88 and 82%. CONCLUSION: Tc-99m-IgG may be usefull in the scintigraphic evaluation of osteomyelitis. PMID- 11022351 TI - [Comparative study of intestinal anastomosis with manual suture and biofragmentable ring in dogs under corticosteroid administration]. AB - BACKGROUND: This study analyzed intestinal anastomoses by manual suture and by compression with biofragmentable ring under delay of cicatrization administering parenteral corticoids. MATERIAL AND METHODS: Twenty dogs were divided into two groups: control and test, the latter submitted to intramuscular administration of hydrocortisone hemisuccinate, 25 to 33 mg/kg/day, on the 30th preoperative and 7th postoperative days. During surgery, each animal underwent two colon sections with anastomosis by manual suture in a single extramucous plane and compression with biofragmentable ring. The animals were sacrificed 7 days after the procedure to evaluate the anastomoses. RESULTS: In the postoperative period, one death occurred in the test group and two in the control group, caused by nonblocked fistula in the anastomoses by ring compression. Statistically, there was a similar incidence of adherences, fistulas, afferent dilatation and obstruction using comparison methods. On microscopy, deficiency in mucous regeneration of the anastomoses by compression was observed. Computerized histological analysis evidenced in the anastomoses by compression, a greater inflammatory reaction, greater edema of the submucous membrane and enlarged scars. CONCLUSIONS: It was concluded that, with the biofragmentable ring, in colonic anastomosis under delay of cicatrization induced by corticoids, similar results to manual suture regarding to postoperative complications incidence were obtained; ring, however, determined worse mucous regeneration and greater cicatricial inflammatory reaction. PMID- 11022352 TI - [Lupus glomerulonephritis: a long-term follow-up]. AB - PURPOSE: To evaluate and the long term course of patients with lupus nephritis, METHOD: Thirty seven patients with lupus nephritis followed in a referral, tertiary care center of a developing country (Brazil) were studied. The length of follow up was 52.4 + 13.3 months and mean age was 26.05 + 11.12 years. 84% of the patients were females and class IV nephritis was found to be the most frequent (80%). RESULTS: At the time of renal biopsy mean serum creatinine was 1.74 + 1.15 mg/dl, and 24 h-proteinuria was 2.62 + 2.89 g. Fifty one per cent of the patients with elevated serum creatinine showed a decrease in these values. Of the variables studied (age, sex, proteinuria, presence of hypertension and serum creatinine at biopsy), serum creatinine elevation was the only one to be associated with poorer prognosis. Remission of the nephrotic syndrome occurred in 65% of the patients. Actuarial survival rate was 96% at 1 year, 82% at 5 years, 70% at 10 years and 70% at 12 years. Five patients developed end stage renal failure and 7 died. Infection was the most frequent(57%) cause of death. CONCLUSION: Among several factors studied the only which has been associated with chronic renal failure was elevated serum creatinine at the time of biopsy. Infections were the main cause of death. PMID- 11022353 TI - [Morphologic changes of the lesser gastric curvature after segmental gastrectomy associated to proximal gastric vagotomy. Experimental study in dogs]. AB - The segmental gastrectomy of the body of the stomach, combined with proximal gastric vagotomy, has been indicated in the treatment of duodenal ulcer, especially in patients with major risk of the recurrent ulceration. PURPOSE: To evaluate the histopathologic changes, secondaries to combined corporeal segmental gastrectomy and proximal gastric vagotomy, in the lesser curvature of the stomach. METHODS: Thirty-six adult mongrel dogs weighing between 10 to 20 kg were divided into three groups of twelve animals each: Group I animals (control) were submitted to laparotomy and gastric manipulation; Group II, animals underwent proximal gastric vagotomy; and Group III, animals underwent corporeal segmental gastrectomy combined with proximal gastric vagotomy. Each group was subdivided into two subgroups of six animals each, and according to the subgroup, the animals were then sacrificed on the third or in the eighth postoperative day. A fragment of the lesser curvature was removed from each stomach for microscopic study. RESULTS: Regarding edema, vascular congestion, inflammatory exudate, fibrosis and cell damage; histopathologic changes found in animals sacrificed in the same postoperative day were slight in Group I, moderate in Group II and intense in the group III. Although nonspecific, the character and site of the lesions were similar to the ones occurring in ischemic process. It was also observed a directly proportional relation between severity of morphologic changes and ischemic potentiality of surgical procedure. CONCLUSION: The segmental gastrectomy probably aggravated the arterial blood supply of the remnant of lesser curvature of the stomach, which was already compromised by devascularization consequent to proximal gastric vagotomy. PMID- 11022354 TI - [Experience in patients with suspected chronic liver disease and contraindication for percutaneous biopsy using modified Ross needle]. AB - PURPOSE: This work aimed at introducing the transjugular liver biopsy technique oriented by radiologic methods in our hospitals. This technique is indicated for obtaining histopathologic diagnosis in patients clinically and laboratory suspected of having chronic liver disease with contraindications to percutaneous liver biopsy. METHODS: Liver biopsy was obtained with a modified Ross needle through the right jugular vein and right hepatic vein under fluoroscopic control. RESULTS: Transjugular liver biopsy was attempted for 39 patients, liver tissue obtained en 32 and histopathologic diagnosis in 25 (64.1%). In 11 patients (28.2%) there was agreement between the diagnoses established before and after biopsy, however, in 14 patients (35.9%), there was disagreement. The yield of diagnosis was low when patients were suspected for cirrhosis. The procedure was well tolerated by the majority of patients. Nonetheless, 1 presented intra abdominal bleeding and required immediate surgery to control retroperitoneal hemorrhage. CONCLUSIONS: Transjugular liver biopsy is useful for the histopathologic diagnosis of patients with chronic liver diseases whenever the percutaneous route is contraindicated. In this series we obtained histopathologic diagnosis for 64.1% of the subjects studied. Patients suspected of having cirrhosis had a low yield of histopathologic diagnosis (50%) when compared to subjects without clinical evidence for cirrhosis (78.9%). The technique is rather complex, and can cause serious complications. This, it should be performed in reference centers in radiology and hepatology. PMID- 11022355 TI - [Natural history of occupational hearing loss induced by noise]. AB - PURPOSE: To evaluate the clinical and audiometric characteristics of occupational hearing loss induced by noise, according to age and time of exposition in years. METHODS: 222 patients with occupational sensorineural hearing loss induced by noise were studied retrospectively, correlating the auditive clinical claims, alterations of audiometric thresholds at frequencies of 250 Hz to 8000 Hz, speech discrimination indicator with age and time of exposure. As a control group were used the audiometric threshold of a population of same medium age, without morbid antecedents of hearing illness, as preconized by ISO 1999 (1990). The group were divided into subgroups and three decades of exposure were analyzed. RESULTS: It was verified that the clinical claims of hipoacusia increases according to the age and time of exposure. The frequency of tinnitus is constant. The audiometric thresholds in the second decade of exposure present variations that depend on the age. The several audiometric curves are parallel, but they are not horizontal. The worst thresholds were found in the high frequencies from 3000 Hz to 8000 Hz, as a clinical and physiopathological consequences of the commitment of basal areas of cochlea. The speech discrimination showed to be worst according to the increase of age and time of exposure. CONCLUSION: Patients with hearing loss disacusia induced by occupational noise present characteristic audiometric thresholds that vary according to age and time of exposure to noise. These characteristics defined and resumed in audiometric curves can constitute a standard of comparison, evaluation and control for exposed populations. PMID- 11022356 TI - [Applicability of the Torrington and Henderson scale]. AB - PURPOSE: To validate the Torrington & Henderson preoperative risk assessment program in the population of surgical patients in an university hospital (Hospital Sao Paulo, UNIFESP, Sao Paulo, Brasil). MATERIAL AND METHODS: We evaluated 1162 patients who underwent to major thoracic, upper and under abdominal surgery using the Torrington & Henderson program. The patients were classified in low (785), moderate (317) and high risk (60). All patients were accompanied daily during the postoperative period by the same medical team which assessed the preoperative period, until discharged or died. RESULTS: The postoperative pulmonary complications were present in 6.1%, 23.3% and 35.0% in the low, moderate and high risk respectively. The relative risk to develop postoperative pulmonary complications was 3.8 higher in the patients with moderate risk and 5.7 higher in the patients with high risk. The mortality rate due to pulmonary complications was 1.7%, 6.3% and 11.7% respectively in the patients with low, moderate and high risk. The relative risk to the death was 3.7 higher in the patients with moderate risk and 6.9 in the high risk. CONCLUSION: We concluded that the Torrington & Henderson preoperative risk assessment program can identify in our population patients who will develop postoperative pulmonary complications. PMID- 11022357 TI - [Airway pressure release in postoperative cardiac surgery in pediatric patients]. AB - OBJECTIVE: Comparison of three modes of MV: intermittent mandatory ventilation with positive end expiratory pressure (IMV + PEEP), APRV and continuous positive airway pressure (CPAP) in children during cardiac surgery post operative with pulmonary hypertension and mild or moderate pulmonary lesion. METHODS: Ten patients were studied with respiratory monitoring (Bear Neonatal Volume Monitor 1) in MV with a continuous flow, time cycled and pressure limited ventilator. The cardiocirculatory variables analyzed were central venous pressure (CVP), oxygen extraction ratio, cardiac rate, systolic arterial pressure, and arterial-mixed venous CO2 difference. Friedman's test (nonparametric) was used to compare the variables in three modalities of ventilation and the Wilcoxon test was used for the variables obtained in two of the modalities. RESULTS: The mean airway pressure (MAP) showed a significant increasing during APRV compared to IMV + PEEP (p = 0.012). The positive inspiratory pressure (PIP), the minute volume and the ratio of oxygen arterial pressure to oxygen inspired fraction (PaO2/FiO2) didn't show statistical difference. During APRV there was a significant decrease in respiratory rate (p = 0.004) and an increase in tidal volume (p = 0.045) when compared to CPAP and IMV + PEEP. In the cardiocirculatory system only CVP showed a significant increased (p = 0.019) during APRV. CONCLUSION: Due to the methodology utilized MAP was higher with APRV resulting in an increased tidal volume without respiratory or cardiocirculatory adverse effects when the three modes were compared. Our results suggest that APRV is a simple and safe method of ventilation. PMID- 11022358 TI - [Bronchofibroscopy in the etiologic diagnosis of lung diseases in patients with acquired immunodeficiency syndrome]. AB - PURPOSE: To evaluate the role of bronchoscopy in diagnosis of pulmonary disease in acquired immunodeficiency syndrome. METHODS: This study analyses a group of 49 patients with AIDS diagnosis admitted at Nereu Ramos Hospital-Florianopolis-SC. It was selected patients with respiratory symptoms and radiologic signs of pulmonary disease, without etiological diagnosis by sputum analysis. These patients were submitted to bronchoscopy with bronchoalveolar lavage, bronchial brush and transbronchial biopsy. The samples were analyzed by BAAR, cytomegalovirus, P. carinii, other fungus and bacterioscopy by Gram. RESULTS: The bronchoscopy was effective on the realization of etiological diagnosis of 71.43% of the cases. The transbronchial biopsy was the most positive technique which confirmed the diagnosis of 59.18% of the cases. The technique with the least positivity was the bronchial brush, with a diagnosis of 4.08% of the cases. The most frequent etiological agent was P. carinii (42.8%), followed by M. tuberculosis (22.86%). No complication was observed in the procedures making. CONCLUSION: The conclusion of this present study was that the bronchoscopy is a secure procedure, with good diagnosis performance for pulmonary disease in this group of patients. PMID- 11022359 TI - Effect of combined treatment with calcitonin on bone densitometry of patients with treated hypothyroidism. AB - INTRODUCTION: Thyroid hormones (TH) may affect bone metabolism and turnover, inducing a loss of bone mass among hyperthyroid and in hypothyroid patients under hormone replacement treatment. Thyroid dysfunction leads to changes in the dynamics of parathyroid hormone (PTH) and calcitonin (CT) secretion. OBJECTIVE: The objective of the study was to determine the usefulness of CT as adjuvant therapy in the prevention of bone loss during the treatment of hypothyroidism. MATERIAL AND METHODS: We studied 16 female patients with recently diagnosed primary hypothyroidism, divided into two groups: group G1 (n = 8) submitted to treatment with thyroxine (L-T4), and Group 2 (n = 8) that, in addition to being treated with L-T4, received a nasal CT spray. All patients were submitted to determination of TSH, free T4, bone mineral densitometry (BMD) and total bone calcium (TBC) at the time of diagnosis, after 6 to 9 months of treatment, and after 12 months of treatment. RESULTS: No statistical significant differences were detected in either group between the total BMD values obtained for the femur and lumbar spine before and after treatment. However, group G1 presented a statistical significant TBC loss after 12 months of treatment compared to initial values. In contrast, no TBC loss was observed in the group treated with LT-4 in combination with CT, a fact that may suggest that CT was responsible for the lower bone reabsorption during treatment of hypothyroidism. PMID- 11022360 TI - [Use of forearm microsurgical flaps for reconstruction in head and neck: experience with 11 cases]. PMID- 11022361 TI - [Brazil 500 years. A Jesuit anatomical nomenclature at the time of discovery]. AB - The purpose of rendering easier for priests to hear confessions from Brazilian indians, in the beginning of the colonization of Brazil by the Portuguese, and in order to serve indians for better communication led Pero de Castilho, a jesuit born in Vila do Esp?rito Santo, to prepare a list of names of parts of the human body. Such a list of tupi (language of the native indians) and portuguese terms of anatomical structures, in alphabetical order, seems to be the first Nomina Anatomica published in Brazil. Such a bilingual vocabulary constitutes a subsidy for the study of both languages spoken soon after the discovery of Brazil and represents a religious related document that contributes to the history of anatomy and medicine. PMID- 11022362 TI - Operationalizing "trance". I: Rationale and research using a psychophenomenological approach. AB - Despite the popularity of the term "trance" among clinicians to describe the subjective effects associated with being hypnotized, heretofore there has been no means to operationalize that definition. The authors present a rationale and psychophenomenological method to operationalize the term "trance" in terms of (a) hypnotic depth, a quantitative measure of subjective trance assessed via a pHGS (predicted Harvard Group Scale) score, derived from regression analysis, and (b) "trance typology profiles," a qualitative differentiation of empirically derived (via cluster and discriminant analyses) categories of subjective trance experiences. The authors then discuss theoretical and clinical implications of this psychophenomenological approach for developing an operational definition of the concept of trance. PMID- 11022363 TI - Hypnosis and distraction differ in their effects on cold pressor pain. AB - On the bases of Hilgard's neodissociation theory and Spano's (1982) sociocognitive theory, volunteers stringently selected for high (N = 10) and low (N = 10) hypnotizability were exposed to a cold pressor pain test during counterbalanced conditions of waking relaxation, distraction, and hypnosis. To better discriminate between hypnosis and distraction conditions, a new distraction procedure was developed involving the memorization of a sequence of colored lights. High hypnotizables showed significantly greater pain relief for hypnosis versus distraction or waking relaxation conditions. High hypnotizables also demonstrated significantly greater pain relief than low hypnotizables in response to hypnosis. Quantitative electroencephalographic (EEG) findings showed significantly greater high theta (5.5-7.5 Hz) activity for highs as compared to lows at parietal (P3) and occipital (01) sites during both hypnosis and waking relaxation conditions. The findings fail to support the socio-cognitive conceptualization of hypnotic behavior while providing additional evidence supporting the neo-dissociation theory and state based theories of hypnosis in general. PMID- 11022364 TI - Enhancing imagery through hypnosis: a performance aid for athletes. AB - This value of imagery in sports is widely acknowledged. The contribution of hypnosis to enhancing athletes' performance is also recognized, but the value of hypnosis in enhancing imagery has little recognition. The reason for this neglect is explored. The study used Martens' Sport Imagery Questionnaire, which asked the participants to image 4 different situations in their own sport--practicing alone, practicing in front of others, watching a teammate, and competing. Participants reported their subjective impression of vividness on four dimensions -visual, auditory, kinesthetic, and affective. The 14 athletes participating imaged each situation in and out of hypnosis--half of the time the imagery in hypnosis came first and half after. The participants reported that the imagery under hypnosis was more intense for each dimension and more intense for each situation. Whether the imagery was done under hypnosis first or after was not significant. The findings suggest that hypnosis substantially enhances imagery intensity and effectiveness. PMID- 11022365 TI - Problems arising from the lack of a standardized method of performing hemoglobin A1c (HbA1c). PMID- 11022366 TI - Influencing care in acute myocardial infarction: a randomized trial comparing 2 types of intervention. AB - The purpose of this study was to evaluate performance feedback delivered by on site presentations compared to mailed feedback on improving acute myocardial infarction (AMI) care. We used a randomized trial including 18 hospitals nested within the Cooperative Cardiovascular Project. Patients comprised AMI Medicare patients admitted before (n = 929, 1994 and 1995) and after intervention (n = 438, 1996). Control hospitals received written feedback by mail. The experimental intervention group received a presentation led by a cardiologist and a quality improvement specialist. We assessed the proportion of patients receiving appropriate AMI care before and after the intervention. Both univariate and multivariate analyses demonstrated no effect of the intervention in increasing the proportion of patients who received reperfusion, aspirin, beta-blockers, or angiotensin-converting enzyme inhibitors. On-site feedback presentations were not associated with a larger improvement in AMI care compared to the mailed feedback. Other interventions, such as opinion leaders and patient-directed interventions, may be necessary in order to improve the care of AMI patients. PMID- 11022367 TI - Using severity-adjusted mortality to compare performance in a Veterans Affairs hospital and in private-sector hospitals. AB - The objective of this study was to compare hospital mortality in Veterans Affairs (VA) and private-sector patients. The study included 5016 patients admitted to 1 VA hospital. Admission severity of illness was measured using a commercial methodology that was developed in a nationwide database of 850,000 patients from 111 private-sector hospitals. The method uses data abstracted from patients' medical records to predict the risk of death in individual patients, based on the normative database. Analyses compared actual and predicted mortality rates in VA patients. VA patients had higher (P < .05) severity of illness than private sector patients. The observed mortality rate in VA patients was 4.0% and was similar (P = .09) to the predicted risk of death (4.4%; 95% confidence interval 4.0-4.9%). In subgroup analyses, actual and predicted mortality rates were similar in medical and surgical patients and in groups stratified according to severity of illness, except in the highest severity stratum, in which actual mortality was lower than predicted mortality (57% vs 73%; P < .001). We found that in-hospital mortality in 1 VA hospital and a nationwide sample of private sector hospitals were similar, after adjusting for severity of illness. Although not directly generalizable to other VA hospitals, our findings nonetheless suggest that the quality of VA and private-sector care may be similar with respect to one important and widely used measure. PMID- 11022368 TI - The use of quality indicators for assessing radical prostatectomy specimens. AB - The information contained in pathology reports of radical prostatectomy specimens is critically important to treating physicians for selecting adjuvant therapy, evaluating therapy, estimating prognosis, and analyzing outcomes. This information is also of importance to patients and their families. In recent years, the Cancer Committee of the College of American Pathologists and the Association of Directors of Anatomic and Surgical Pathology developed suggested protocols for reporting the findings on radical prostatectomy specimens. The objectives of this study were to assess radical prostatectomy-specimen reports by using quality indicators derived from existing suggested protocols and to thereby assist pathologists in improving the quality of their reports on such specimens. A retrospective chart review of 554 cases for the second 6-month period of 1996 focused on 10 quality indicators: submission of a frozen section; location of the adenocarcinoma; proportion of the specimen involved by adenocarcinoma; perineural involvement; vascular involvement; seminal vesicle involvement; periprostatic fat status; number of nodes submitted; status of nodes; and prostate intraepithelial neoplasia (PIN). The findings of this study were shared with the pathology departments in all hospitals in New York State. In addition, the 113 hospitals from which the 554 cases were drawn were given their institution-specific data. Teleconferences were held with the 37 hospitals that accounted for 72.4% of all cases. These conferences included directors of pathology and laboratories and focused on the aggregate statewide findings. The presence of quality indicators in reports varied from a mean of 14.8% (periprostatic fat) to a mean of 85.9% (seminal vesicle involvement). For all hospitals, 4 indicators (proportion of the specimen involved by adenocarcinoma, vascular involvement, periprostatic fat status, and PIN) were included in fewer than 50% of cases. These 4 quality indicators and an additional 3 others (submission of a frozen section, perineural involvement, and the number of nodes submitted) were included in fewer than 70% of cases. Only 3 indicators (location of the adenocarcinoma, seminal vesicle involvement, and the status of nodes) were found in more than 70% of cases. Although the mean level of quality indicator inclusion ranged from 14.8% to 85.9% for all cases examined, the absolute range for any individual indicator was 0% to 100%. Thus, some hospitals included a given indicator 100% of the time; others never included it. This pattern held true for all 10 indicators. High-volume hospitals (10 or more cases) performed significantly better than low-volume hospitals (1-4 cases) on 5 indicators (P < .05), and better, but not significantly so, for an additional 2 indicators. Overall, the mean inclusion levels for all 10 indicators were 10% higher for high-volume hospitals compared with low-volume and medium-volume hospitals (5-9 cases). This study demonstrated wide variations in the inclusion of quality indicators by pathologists in their radical prostatectomy-pathology reports. Whereas some hospitals always include given indicators, others never mentioned them. These marked disparities point to the need for standardized reporting for radical prostatectomy specimens. PMID- 11022369 TI - A quality improvement and risk management initiative for surgical ICU patients: a study of the effects of physical restraints and sedation on the incidence of self extubation. PMID- 11022370 TI - Whither.com. PMID- 11022371 TI - Sensationalist claims unprofessional. PMID- 11022372 TI - Supernumerary upper anterior teeth. PMID- 11022373 TI - Quality assurance--are we really ready? PMID- 11022374 TI - Inductive vs deductive reasoning. PMID- 11022375 TI - Feedback not so NICE. PMID- 11022376 TI - Feedback not so NICE. PMID- 11022377 TI - Feedback not so NICE. PMID- 11022379 TI - Fluorides and the prevention of dental decay: a statement from the Representative Board of the British Dental Association. AB - Fluoride reduces the incidence of dental decay substantially. Fluoride benefits all age-groups, because of its topical effect. Water fluoridation needs to be targeted on high caries areas which are normally also areas of social and material deprivation. Thorough brushing twice a day with a fluoride toothpaste is also crucial to the improvement of oral health. PMID- 11022378 TI - The Eastman Jubilee 1948-1998. AB - The postgraduate Eastman Dental Clinic and Institute of Dental Surgery were established in 1947. However their roots go back to Rochester in New York State where George Eastman was born in 1854. George Eastman literally changed the image of the world by creating popular photography. In 1914 he was driven by his painful neglected dentition to seek the help of Harvey Jacob Burkhart (pictured, right) who was considered to be the best dentist in the state. Burkhart provided him, for the first time in his adult life, with comfortable dentures, which not only restored function but also his appearance. George Eastman was overwhelmed with his new found comfort and social confidence. PMID- 11022380 TI - Tongue piercing resulting in hypotensive collapse. AB - Tongue piercing remains popular. A variety of complications have been reported, including life-threatening infection, airway problems and damaged teeth or mucosal surfaces. A patient who collapsed after continuous profuse bleeding following tongue piercing is presented. It is recommended that piercing practitioners be licensed and inspected. A list of written post piercing instructions for customers is included on how to deal with, or who to contact regarding potential complications including haemorrhage. PMID- 11022381 TI - Registration: stage III--selection of teeth. PMID- 11022382 TI - Antibiotic prescribing by general dental practitioners in the Greater Glasgow Health Board, Scotland. AB - OBJECTIVE: To investigate antibiotic prescribing patterns by general dental practitioners (GDPs) in the Greater Glasgow Health Board Area, Scotland. STUDY DESIGN: A 10% sample of prescriptions were selected at random from 35,545 prescriptions written by GDPs over a 6-month period. MAIN OUTCOME MEASURES: Absolute and relative frequencies were used to describe the different classes of antibiotics used and the variations in prescribing practice. RESULTS: GDPs prescribed a wide range of antibiotics. Seventeen different antibiotics were prescribed with amoxycillin, metronidazole and penicillin V accounting for almost 90% of the prescriptions. In general the antibiotics were prescribed at the British National Formulary (BNF) recommended doses. There were, however, wide variations in the frequencies and durations of the prescriptions for all antibiotics. CONCLUSIONS: The present study provides evidence of sub-optimal prescribing of antibiotics by dentists in Scotland, with considerable variation from the recommended frequencies and doses. PMID- 11022383 TI - A clinical evaluation of an Erbium:YAG laser for dental cavity preparation. AB - OBJECTIVE: A randomised controlled trial to determine the acceptability to dentists and patients of cavity preparation with an Erbium:YAG laser as compared with conventional handpieces. METHODS: Fifteen dentists (9 GDPs, 1 community dentist and 5 hospital dentists) treated 77 patients (age range 3.5-68 years old) who had two matched cavities, in a split mouth, randomised trial. One cavity was prepared conventionally, the other with the laser, with dentist and patient preference determined by questionnaire. RESULTS: In the majority of cases, where dentists expressed a preference, it was for conventional cavity preparation, and this was significant (P < 0.001). In more than half of the laser appointments, dentists had to use conventional handpieces to complete the cavity. Principle difficulties reported with the laser were access (25 cases) and slow speed of cutting (11 cases). Patients aged > or = 10 years who expressed a preference, preferred laser treatment, and this was significant (P < 0.001). Patients aged < 10 years, assessed using a simplified pictorial questionnaire, did not show a significant preference for either technique. CONCLUSIONS: Dentists preferred conventional handpieces for cavity preparation while patients aged > or = 10 years old preferred laser treatment. Patients < 10 years old did not express a preference. PMID- 11022384 TI - High and low torque handpieces: cutting dynamics, enamel cracking and tooth temperature. AB - OBJECTIVE: The aim of these experiments was to compare the cutting dynamics of high-speed high-torque (speed-increasing) and high-speed low-torque (air-turbine) handpieces and evaluate the effect of handpiece torque and bur type on sub surface enamel cracking. Temperature changes were also recorded in teeth during cavity preparation with high and low torque handpieces with diamond and tungsten carbide (TC) burs. The null hypothesis of this study was that high torque handpieces cause more damage to tooth structure during cutting and lead to a rise in temperature within the pulp-chamber. MATERIALS AND METHODS: Images of the dynamic interactions between burs and enamel were recorded at video rate using a confocal microscope. Central incisors were mounted on a specially made servomotor driven stage for cutting with a type 57 TC bur. The two handpiece types were used with simultaneous recording of cutting load and rate. Sub-surface enamel cracking caused by the use of diamond and TC burs with high and low torque was also examined. Lower third molars were sectioned horizontally to remove the cusp tips and then the two remaining crowns cemented together with cyanoacrylate adhesive, by their flat surfaces. Axial surfaces of the crowns were then prepared with the burs and handpieces. The teeth were then separated and the original sectioned surface examined for any cracks using a confocal microscope. Heat generation was measured using thermocouples placed into the pulp chambers of extracted premolars, with diamond and TC burs/high-low torque handpiece variables, when cutting occlusal and cervical cavities. RESULTS: When lightly loaded the two handpiece types performed similarly. However, marked differences in cutting mechanisms were noted when increased forces were applied to the handpieces with, generally, an increase in cutting rate. The air turbine could not cope with steady heavy loads, tending to stall. 'Rippling' was seen in the interface as this stall developed, coinciding with the bur 'clearing' itself. No differences were noted between different handpieces and burs, in terms of sub-surface enamel cracking. Similarly, no differences were recorded for temperature rise during cavity preparation. CONCLUSIONS: Differences in cutting mechanisms were seen between handpieces with high and low torque, especially when the loads and cutting rates were increased. The speed increasing handpiece was better able to cope with increased loading. Nevertheless, there was no evidence of increased tooth cracking or heating with this type handpiece, indicating that these do not have any deleterious effects on the tooth. PMID- 11022385 TI - So the difference is what? PMID- 11022386 TI - Leptospirosis outbreak in Eco Challenge 2000 participants. PMID- 11022387 TI - Influenza activity updates 2000/2001. PMID- 11022388 TI - Three nursing home outbreaks of Norwalk-like virus in Brisbane in 1999. AB - We report on three nursing home outbreaks of gastroenteritis in Brisbane in 1999. The presence of Norwalk-like virus (NLV) genogroup 2 was demonstrated by reverse transcription polymerase chain reaction (RT-PCR) in all three outbreaks. Common findings of these investigations were rapid spread of the illness within the institutions and difficulties in identifying a common source. Nursing home populations are vulnerable and it is important for each institution to have infection control policies in place so outbreaks can be managed promptly. This includes the exclusion of ill staff for 48 to 72 hours after recovery from illness. Genogrouping of NLV by RT-PCR can take several days so control measures will generally have to be instituted before results become available. PMID- 11022389 TI - Typhoid and paratyphoid fever in south-eastern Sydney, 1992-1997. AB - Notification records of typhoid and paratyphoid cases among residents of south eastern Sydney during 1992-1997 were reviewed, with particular attention paid to identifying a source of infection and to completeness of follow up. Notifications comprised 30 cases of Salmonella Typhi, nine of S. Paratyphi A and five of S. Paratyphi B. These 44 cases had a median age of 20 years (range 2-62). Of the 39 cases with known country of birth, 30 were born overseas, predominantly in Asian countries. Of 39 cases with a known travel history, 33 were cases of overseas acquired acute infection and two cases were asymptomatic chronic carriers. A source was identified in only one of four domestically acquired infections. Of eight household contacts in occupations posing a public health risk (seven food handlers and one health-care worker), complete follow-up information was available for only five. Most cases were in overseas-born individuals who may have been infected when returning to their country of birth. Explicit follow-up protocols need to cover appropriate clinical management (including treatment of chronic carriage) and monitoring of those cases and contacts who could pose a public health risk. PMID- 11022390 TI - A community outbreak of Cryptosporidium infection associated with a swimming pool complex. AB - A case-control study was conducted to investigate the cause of a sudden increase in cases of cryptosporidiosis notified to the Brisbane Southside Public Health Unit from January to March 1998. Fifty-two eligible cases were identified over a three-week period early in 1998. Thirty-one of these cases and 21 control subjects participated in the study. Swimming in the 2 weeks before onset of illness was identified as a likely risk factor for cryptosporidiosis infection (OR 3.1, CI 0.8-12.6, P = 0.06). Analysis of swimming pool attendance identified swimming at Pool Complex A as a significant risk factor for the acquisition of cryptosporidiosis (OR 8.9, CI 1.5-67.4, P = 0.004). No other potential risk factors were significantly associated with illness. The detection of cryptosporidium oocysts in three of the four pools at Pool Complex A supported the findings of the case-control study. As a response to this outbreak, Queensland Health has developed a Code of Practice outlining measures for the control and prevention of future outbreaks of swimming pool-associated cryptosporidiosis and/or giardiasis. PMID- 11022391 TI - Notifications of vaccine preventable diseases in Australia. Quarterly report (January-March 2000). AB - Vaccine preventable disease notifications for Australia with disease onset dates between January and March 2000 are reviewed. During this quarter, numbers of notifications for Haemophilus influenzae type b disease and measles were the lowest ever recorded, while those for rubella were the lowest recorded since before the epidemic of spring 1992. These are promising trends that are likely to represent a true reduction in disease incidence. Numbers of pertussis notifications declined compared with the last quarter of 1999, but remain high, making up 88% of notifications for the vaccine preventable diseases reported here. PMID- 11022393 TI - Communicable Diseases Surveillance. Presentation of NNDSS data. PMID- 11022392 TI - Young adult measles vaccination. PMID- 11022394 TI - Immunisation coverage estimates. PMID- 11022395 TI - Sex-related differences: do they matter? PMID- 11022396 TI - Influence of chronic barbiturate administration on sleep apnea after hypersomnia presentation: case study. AB - When sleepiness is excessive, undesirable, inappropriate or unexplained, it often indicates a clinical disorder that is generically termed hypersomnia. One of the leading causes of hypersomnia is sleep apnea. We present the case of a 44-year old woman with a history of bipolar spectrum disorder and epilepsy who initially showed evidence of hypersomnia. The hypersomnia settled with changes to her medication, but the patient was subsequently found to have severe obstructive sleep apnea. The relation between the patient's medication and sleep apnea is discussed, and the possible respiratory-suppressant effects of chronic barbiturate treatment are considered. The role of other evoking factors within the context of this case and the mechanisms by which drug interactions and psychotropic treatment may worsen, obscure or perpetuate sleep apnea are also examined. PMID- 11022397 TI - Probing peripheral and central cholinergic system responses. AB - OBJECTIVE: The pharmacological response to drugs that act on the cholinergic system of the iris has been used to predict deficits in central cholinergic functioning due to diseases such as Alzheimer's disease, yet correlations between central and peripheral responses have not been properly studied. This study assessed the effect of normal aging on (1) the tropicamide-induced increase in pupil diameter, and (2) the reversal of this effect with pilocarpine. Scopolamine was used as a positive control to detect age-dependent changes in central cholinergic functioning in the elderly. DESIGN: Randomized double-blind controlled trial. PARTICIPANTS: Ten healthy elderly (mean age 70) and 9 young (mean age 33) volunteers. INTERVENTIONS: Pupil diameter was monitored using a computerized infrared pupillometer over 4 hours. The study involved 4 sessions. In 1 session, tropicamide (20 microL, 0.01%) was administered to one eye and placebo to the other. In another session, tropicamide (20 microL, 0.01%) was administered to both eyes, followed 23 minutes later by the application of pilocarpine (20 microL, 0.1%) to one eye and placebo to the other. All eye drops were given in a randomized order. In 2 separate sessions, a single dose of scopolamine (0.5 mg, intravenously) or placebo was administered, and the effects on word recall were measured using the Buschke Selective Reminding Test over 2 hours. OUTCOME MEASURES: Pupil size at time points after administration of tropicamide and pilocarpine; scopolamine-induced impairment in word recall. RESULTS: There was no significant difference between elderly and young volunteers in pupillary response to tropicamide at any time point (p > 0.05). The elderly group had a significantly greater pilocarpine-induced net decrease in pupil size 85, 125, 165 and 215 minutes after administration, compared with the young group (p < 0.05). Compared with the young group, the elderly group had greater scopolamine-induced impairment in word recall 60, 90 and 120 minutes after administration (p < 0.05). CONCLUSION: There is an age-related pupillary response to pilocarpine that is not found with tropicamide. Thus, pilocarpine may be useful to assess variations in central cholinergic function in elderly patients. PMID- 11022399 TI - Effects of clozapine and typical antipsychotic drugs on plasma 5-HT turnover and impulsivity in patients with schizophrenia: a cross-sectional study. AB - OBJECTIVE: To compare the efficacy of clozapine with typical antipsychotic drugs in controlling impulsivity and to explore the possible correlation of impulsivity with plasma 5-hydroxytryptamine (5-HT) levels, plasma 5-hydroxyindoleacetic acid (5-HIAA) levels and plasma 5-HT turnover. DESIGN: Prospective, cross-sectional study open to medication and blinded to biochemical analyses. PARTICIPANTS: Healthy control subjects (n = 24) and 46 inpatients and outpatients meeting the DSM-IV criteria for schizophrenia; 20 were being treated with clozapine and 26 were taking typical antipsychotic drugs. INTERVENTIONS: All psychotropic drugs other than clozapine or typical antipsychotic drugs were discontinued for at least 5 days and subjects fasted overnight before they were assessed. OUTCOME MEASURES: Coccaro Impulsivity Scale scores, plasma 5-HT levels, 5-HIAA levels and 5-HT turnover. RESULTS: Patients treated with clozapine and those treated with typical antipsychotics had significantly higher impulsivity scores than the control group, and the mean impulsivity score of the typical antipsychotic group was significantly higher than that of patients treated with clozapine. The mean concentration of 5-HT of the typical antipsychotic group was significantly lower than that of the control group and patients treated with clozapine; however, mean plasma levels of 5-HIAA were significantly higher for the clozapine group than the other 2 groups. 5-HT turnover was significantly higher for the 2 drug treatment groups than for the control group. CONCLUSIONS: These results suggest that treatment with clozapine should be considered for patients with schizophrenia who are impulsive and aggressive. PMID- 11022398 TI - Preliminary randomized double-blind placebo-controlled trial of tryptophan combined with fluoxetine to treat major depressive disorder: antidepressant and hypnotic effects. AB - OBJECTIVE: Because the initial phase of treatment of depression with a selective serotonin reuptake inhibitor is often complicated by a delayed onset of action of the antidepressant or severe insomnia or both, we investigated whether tryptophan, an amino acid with both antidepressant-augmenting and hypnotic effects, would benefit patients with depression at the beginning of treatment with fluoxetine. DESIGN: Randomized, double-blind, placebo-controlled trial. PATIENTS: Thirty individuals with major depressive disorder. INTERVENTIONS: Treatment over 8 weeks with 20 mg of fluoxetine per day and either tryptophan (2 to 4 g per day) or placebo. OUTCOME MEASURES: Mood was assessed using the 29-item Hamilton Depression Rating Scale (HDRS-29) and the Beck Depression Inventory (BDI). Laboratory sleep studies were done at baseline and after 4 and 8 weeks of treatment using standard procedures. RESULTS: During the first week of treatment, there was a significantly greater decrease in HDRS-29 depression scores, and a similar trend in BDI scores, in the tryptophan/fluoxetine group than in the placebo/fluoxetine group. No significant differences were noted at later time points. With respect to sleep measures, there was a significant group-by-time interaction for slow-wave sleep at week 4. Further analysis revealed a significant decrease in slow-wave sleep after 4 weeks of treatment in the placebo/fluoxetine group, but not in the tryptophan/fluoxetine group. No cases of serotonin syndrome occurred, and the combination was well tolerated, although the 4 g per day dosage of tryptophan produced daytime drowsiness. CONCLUSIONS: Combining 20 mg of fluoxetine with 2 g of tryptophan daily at the outset of treatment for major depressive disorder appears to be a safe protocol that may have both a rapid antidepressant effect and a protective effect on slow-wave sleep. Further large-scale studies are needed to confirm these initial findings. PMID- 11022400 TI - Association and linkage studies of candidate genes involved in GABAergic neurotransmission in lithium-responsive bipolar disorder. AB - OBJECTIVE: To test for genetic linkage and association with GABAergic candidate genes in lithium-responsive bipolar disorder. DESIGN: Polymorphisms located in genes that code for GABRA3, GABRA5 and GABRB3 subunits of the GABAA receptor were investigated using association and linkage strategies. PARTICIPANTS: A total of 138 patients with bipolar 1 disorder with a clear response to lithium prophylaxis, selected from specialized lithium clinics in Canada and Europe that are part of the International Group for the Study of Lithium-Treated Patients, and 108 psychiatrically healthy controls. Families of 24 probands were suitable for linkage analysis. OUTCOME MEASURES: The association between the candidate genes and patients with bipolar disorder versus that of controls and genetic linkage within families. RESULTS: There was no significant association or linkage found between lithium-responsive bipolar disorder and the GABAergic candidate genes investigated. CONCLUSIONS: This study does not support a major role for the GABAergic candidate genes tested in lithium-responsive bipolar disorder. PMID- 11022401 TI - Statistical mapping analysis of serotonin synthesis images generated in healthy volunteers using positron-emission tomography and alpha-[11C]methyl-L-tryptophan. AB - OBJECTIVES: To assess the suitability of analyzing functional images of brain serotonin (5-HT) synthesis with statistical parametric mapping (SPM), and to investigate further possible sex-related regional differences. DESIGN: Prospective study. PARTICIPANTS: Six healthy men and 5 healthy women. INTERVENTION: Participants' brains were scanned with positron-emission tomography (PET) after intravenous injection of alpha-[11C]methyl-L-tryptophan (alpha [11C]MTrp). OUTCOME MEASURES: Tissue radioactivity images were converted into functional images using the Patlak plot approach, and analyzed with 2 methods for global normalization in the SPM program: proportional scaling and analysis of covariance (ANCOVA). RESULTS: The data structure suggests that PET alpha [11C]MTrp data meet the criteria for analysis with SPM, and that the proportional scaling method is more appropriate than the ANCOVA method for normalization. Regional differences in 5-HT synthesis were identified between men and women, and the significance of these findings was supported by region of interest (ROI) analyses. CONCLUSION: SPM analyses of PET alpha-[11C]MTrp data may be of value for identifying regional differences in brain 5-HT synthesis between groups, and in investigating the effects of psychotropic drugs. Since we found regional differences between male and female subjects, men and women should not be grouped for data analysis in PET alpha-[11C]MTrp studies. PMID- 11022404 TI - Should we encourage the use of high-dose vitamin E in persons with memory complaints as a preventive strategy against Alzheimer's disease? PMID- 11022402 TI - Relation of serum cholesterol, lipid, serotonin and tryptophan levels to severity of depression and to suicide attempts. AB - OBJECTIVE: To determine if there is a relation to low serum cholesterol, lipoprotein, serotonin or tryptophan levels in patients with depression who have recently attempted suicide. DESIGN: Biochemical and behavioural study. SETTING: Inpatient and outpatient treatment at the Instituto Mexicano de Psiquiatria. PARTICIPANTS: Thirty-three patients with a diagnosis of major depressive episode. Eighteen of these patients had attempted suicide in the month before the start of the study; 15 patients had never attempted suicide. OUTCOME MEASURES: Serum levels of total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides, serotonin (5-HT) and tryptophan. Scores on Hamilton Depression Rating Scale, Carroll Depression Rating Scale, Beck Hopelessness Scale and Beck Suicide Attempt Severity Scale. RESULTS: There were no significant differences between patients who had attempted suicide and those who had not in terms of serum cholesterol, HDL, LDL and triglyceride levels. Serum levels of 5-HT and tryptophan were significantly lower in patients with depression who had a recent suicide attempt than in those patients who had never attempted suicide. A comparison of patients not taking antidepressant medication found serum 5-HT levels to be more than 3 times lower in those patients with a recent suicide attempt than in patients with no history of suicide attempt. CONCLUSIONS: The study found no difference in lipid profiles between patients who had attempted suicide and those who had not. Low serum levels of 5-HT may increase the risk of suicide attempt in patients who are depressed. PMID- 11022406 TI - Transmyocardial revascularization--late results and mechanisms of action. AB - Transmyocardial revascularization (TMR) has been used in over 500 patients to relieve severe angina when all other measures failed: two different lasers were used in the study. Each has been successful, but, in the author's experience, the Carbon Dioxide laser has given better relief of angina and increase in perfusion than the Holmium-YAG laser. Based on these clinical observations, the probable mechanism of action is stimulation of vascular neogenesis plus improved distribution of the available blood supply. PMID- 11022405 TI - Cervical cancer in Kentucky. PMID- 11022403 TI - Effects of sustained (+/-)pindolol administration on serotonin neurotransmission in rats. AB - OBJECTIVE: Given reports that (+/-)pindolol, a beta-adrenergic-5-HT1A/1B receptor antagonist, accelerates the onset of the therapeutic effect of certain antidepressant drugs in major depression, the aim of this study was to assess the effect of sustained (+/-)pindolol administration on the sensitivity of pre- and postsynaptic 5-HT1A receptors, terminal 5-HT1B autoreceptors and on overall 5-HT neurotransmission. DESIGN: Prospective animal study. ANIMALS: Sprague-Dawley rats. OUTCOME MEASURES: Modifications of the sensitivity of somatodendritic and postsynaptic 5-HT1A receptors using in vivo electrophysiological paradigms in animals treated with vehicle or (+/-)pindolol (20 mg/kg/day, subcutaneously) through osmotic minipumps for 2 weeks. RESULTS: (+/-)Pindolol attenuated the suppressant effect of the 5-HT autoreceptor agonist lysergic acid diethylamide (LSD) on the firing activity of 5-HT neurons, suggesting that (+/-)pindolol antagonized somatodendritic 5-HT1A autoreceptors in the dorsal raphe nucleus. However, following a 2-day washout period, the suppressant effect of LSD was still attenuated, indicating rather a desensitization of 5-HT1A autoreceptors had occurred. In the CA3 region of the dorsal hippocampus, (+/-)pindolol treatment did not modify the responsiveness of postsynaptic 5-HT1A receptors to microiontophoretic applications of 5-HT. Moreover, such a treatment modified neither the effectiveness of the electrical stimulation of 5-HT fibers nor the function of terminal 5-HT autoreceptors. Finally, the administration of the selective 5-HT1A receptor antagonist WAY 100635 (100 micrograms/kg, intravenously) did not increase the firing activity of dorsal hippocampus CA3 pyramidal neurons in rats treated with (+/-)pindolol, thus failing to reveal the enhanced tonic activation of postsynaptic 5-HT1A receptors associated with major classes of antidepressant treatments. CONCLUSION: Prolonged administration of (+/ )pindolol by itself is not sufficient to enhance overall 5-HT neurotransmission; pindolol should therefore not be endowed with intrinsic antidepressant activity. Although pindolol is capable of antagonizing the 5-HT1A autoreceptor upon the initiation of a 5-HT reuptake-blocker treatment, it also induces a desensitization of this 5-HT1A autoreceptor, which could explain why patients do not relapse upon its discontinuation when they continue taking a 5-HT reuptake blocker. PMID- 11022407 TI - [Regulatory assessment of the "Nutricion Hospitalaria" journal]. AB - A study is carried out to determine the degree of compliance by the journal entitled "Nutricion Hospitalaria" with the international standards issued by the ISO (International Standards Organization) by means of the application of a survey questionnaire drawn up in accordance with the guidelines published by national and international standards organizations. In total, 161 parameters are considered. Together with the general aspects of the publication, an analysis is made of its specific sections, decisive in the establishment of its level of compliance. After the analysis carried out, it was seen that the general standardization mean was 64% and the real average was 76.82%. At the same time, the necessary recommendations are given to improve these standardization levels and improve the editorial quality of "Nutricion Hospitalaria". PMID- 11022408 TI - [Oxidation of lipids contained in total parental nutrition]. AB - The components used in parenteral nutrition mixtures are subject to oxidation. The oxidation of polyunsaturated lipids leads to the formation of toxic lipoperoxides. This toxicity is of particular significance in new-born children, because of the lack of maturity of their defensive systems against oxidative processes, and also in critically-ill patients, whose oxidative metabolism is increased. A review of the literature is effected to assess the following: toxicity of lipoperoxides, factors involved in the oxidation of parenteral nutrition mixtures and the analytical methods used to determine the peroxides present in parenteral nutrition mixtures. The factors influencing the oxidation of parenteral nutrition mixtures include: the amount of polyunsaturated fatty acids, the presence of bisulphite, vitamin complexes and their polysorbate content, temperature, type of bag and exposure to light. The analytical methods found for the analysis of peroxides in TPN mixtures are: iodine measurement analysis, high resolution liquid chromatography, the thiobarbituric acid test, and oxidation of the ferrous cation in the presence of xylenol orange. Depending on the method, it is possible to determine total peroxides or only lipoperoxides. The literature reviewed does not establish a threshold level of peroxide concentration below which a parenteral nutrition mixture can be considered safe. PMID- 11022409 TI - [Status of feeding services at Spanish hospitals with dietetic and clinical nutrition unit]. AB - Nutrition at the hospital is substantial to the evolution of the disease. The number of diets in the surveyed hospital ranged between 15 and 70 and was not influenced by the hospital number of beds. The proportion of basal and therapeutic diets is similar (about 50% each), although the number of "special" diets increase with the hospital size. The average periodicity of menus is 14 days, while the optional "menu" is lower in bigger hospitals. The knowledge of the Diet Code in different hospitals is how between physicians and nurses in relation to other health and service professionals. The computerized ordering of diets is higher in those hospitals with more than 1500 beds (66%) and the manual request ranges between 65% and 80% in those with 200 to 1500 beds. Most hospitals have a centralised distribution system with a isothermal tray. PMID- 11022410 TI - [Zinc intake deficiency and colorectal cancer: what is the situation in our population?]. AB - INTRODUCTION AND OBJECTIVES: Various experimental studies have shown that a deficient intake of zinc increases the proliferation of cells, the effect of carcinogens and the progression of colonic adenomas to carcinomas. The purpose of our study is to determine whether this relationship between zinc intake deficit and colorectal cancer can be found in our population. METHODS: The study included 100 cases of colorectal cancer which were compared with 200 control patients (76% hospital admissions, 22% in the community and 2% with normal colonoscopy). The sampling of cases was done consecutively without use of probabilities and with the later application of a stratified matching design. For data collection purposes, a food survey form was used. For the determination of a deficient zinc intake this was compared with the values of the United States' Recommended Dietary Allowance (RDA). Deficient zinc intake was defined as values lower than 75% of RDA. Statistical calculations were effected by calculating chi-squared. The level of statistical significance was set at p < 0.05. RESULTS: Seventy-nine patients with cancer (79%) presented zinc intake deficiency, whereas the figure for the control group was 142 individuals (71%). From these data, a chi-square obtained was 2.19, corresponding to a p > 0.1. CONCLUSIONS: Despite the evidence of the experimental studies, we have been unable to identify any relationship between deficient zinc intake and colorectal cancer in our population. PMID- 11022411 TI - [Assessment of nutritional status at hospital admission: identification of patients with risk for malnutrition]. AB - Various studies of prevalence in our hospitals have detected large percentages of under-nourished patients in both medical and surgical wards. These groups of patients are not detected in clinical practice and are therefore untreated, leading to nutritional deterioration. The present paper studies the prevalence of the risk of malnutrition, using the Cardona risk assessment sheet as modified by Mager, in 134 patients between the 3rd and 5th day after admission to hospital. A chi-squared test was applied to allow comparison of quantitative and qualitative variables. We found 56.70% of patients risked under-nourishment (60% in medical wards and 50% in surgical wards). The variables which showed the best correlation with the risk of malnutrition were albumen (p < 0.01), weight loss (p < 0.001) and age (p < 0.005). The Hospital's Clinical Nutrition Committee must define the standard to be achieved regarding nutritional risk in order to ensure the quality offered to our patients in this regard. PMID- 11022412 TI - [Manufacturing a double-lumen tube with simple and inexpensive materials]. AB - Twin-channel enteral feeding catheters allow the administration of early enteral nutrition in various pathologies while avoiding the use of total parenteral nutrition. We describe how to manufacture one simply using cheap and easy-to-find materials whenever a commercial catheter of this type is unavailable. PMID- 11022413 TI - [Fundamental bases of biological rhythms]. AB - The data and theoretical points given in the paper mould basically new views of molecular relationships underlying the function of living beings and biological rhythms. The authors' procedure for wedge biological fluid dehydration reveals a wide autowave spectrum that is clearly detectable when the fluid passes into the solid phase. A hypothesis of the autowave interaction of biologically active molecules is forwarded, which considers autowaves as a basis of organization of physiological and pathological processes occurring in the body. PMID- 11022414 TI - [From chronobiology to chronotopobiology]. AB - Based on the data available in the literature and his own findings, the author presents a concept of the spatial-temporary organization of biological systems. Experimental evidence is provided for this concept with emphasis on its theoretical and medical values. A role of spatial and temporary organization in the activity of the biological system is shown separately. It is emphasized that the spatial-temporary organization of biological system is not the sum of properties of components of its spatial and temporary organizations and has its own features. The author proposed to single out a section of theoretical biology dealing with the study of the spatial-temporary organization of biological systems and to call it chronotopobiology. PMID- 11022415 TI - [Diurnal rhythm of sensitivity of nerve cell genome activity to changed motor regime]. AB - Diurnal variations in the incorporation of labelled precursors of RNA and proteins into the spinal, stem, and cerebral cell populations of the rat nervous system were studied on surviving brain slices under standard conditions and during exercise by swimming. Cosinor analysis of the relative level of 3H-uridine incorporation into spinal ganglia L5, superior cervical ganglia, spinal cord L5, hypothalamic suprachiasmatic nuclei and cerebellar cortex revealed that the arrangement of the calculated acrophases of these organs in the dark period of a day. The acrophases of visual and somatosensory cortical cells were observed in the light period of a day. The calculated maximum values of the rate of 3H leucine incorporation into nerve cell proteins (the internal acrophases ranged from 3 to 9 hours) were seen at certain intervals. Thus, the processes providing the function of a protein-synthesizing apparatus of nerve cells are characterized by a definite time organization and these processes vary significantly in their activity during a diurnal cycle. Depending on the time of a day when exercise was performed, there were differences in the changing activity of genetic and protein synthesizing apparatuses during nerve cell reactions. This is associated with the different time of the onset of a reaction, passage of its some stages, with the rate of processes, and with the periods of normalization. The curves of a phasic response are discussed. PMID- 11022416 TI - [Brain rhythm-organizing structures and pharmacological effect]. PMID- 11022417 TI - [Ulcer protective effect of melatonin during artificial simulation of desynchronization in rats]. AB - The study was made on Wistar rats. A shift in endogenous biological rhythms was simulated by that in the light phase by 12 hours and evaluated by the shift in the rhythm of rectal temperature. Experiments revealed that desynchronization potentiated ulcerative processes in the rat stomach. The study demonstrated the protective effect of intraperitoneally injected melatonin to prevent the formation of ulcerative defects in the rat stomach during artificially simulated desynchronization. PMID- 11022418 TI - [Circadian variability of cardiac arrhythmias in patients with cardiovascular diseases]. AB - A total of 1063 results of Holter monitoring in patients with cardiovascular diseases were analyzed. There were regularities in the circadian rhythm of arrhythmias with their decreased values at night and increased ones in the daytime, and peak in the morning. The profile of rhythm depends on the pattern of arrhythmia and cardiac disease. The main oscillators are as follows: sleep awakening, activity-rest patterns, circadian variations in the sympathetic tone, severity of disease, diurnal variations in myocardial oxygen demand, blood pressure, cardiosclerosis. Conduction disturbances were increased during sleep and early in the morning. Thus, the main unfavourable period in arrhythmia disturbances are early morning, pre- and postawakening periods. PMID- 11022419 TI - [Effects of biorhythm regulator melatonin on DNA synthesis in short-term cultures of human malignant tumors]. AB - The cytotoxic action of melatonin on DNA synthesis was studied in short-term human tumor tissue cultures. Twenty tumors (ovarian, renal, colorectal, gastric, skin, testicular, thyroid, adrenal gland, endometrial, cervical uterus ones, and melanoma) were isolated from patients at surgery. The effect of melatonin (concentrations of 5 x 10(-5) M to 5 x 10(-13) M on the level of 3H-thymidine incorporation into tumor tissue DNA (4-hour exposure) was evaluated. After exposure, 3H-thymidine was added to the medium for an hour. The quantity of DNA was determined in the hydrolysate by spectrometry; the level of 3H-thymidine incorporation into DNA was radiometrically evaluated by a scintillation counter. The quantity corresponding to 3H-thymidine incorporation per unit of DNA was determined. The control group and the melatonin-treated groups were compared by ANOVA. Melatonin inhibited DNA synthesis in 11 (55%) of the 20 tumors and was ineffective in 9 (45%) of the 20 ones. Melatonin-sensitive tumors were as follows: endometrial CA (33% vs 75% inhibition, p < 0.01), gastric CA (71% inhibition, p < 0.05), adrenal gland CA (38% inhibition, p < 0.01). Ovarian, cervical, skin CA and melanoma were unresponsive to melatonin. Renal cell CA, colonic and rectal CA were sensitive in some cases. In sensitive tumors, melatonin was effective even at concentrations of 5 x 10(-13) M to 5 x 10(-7) M. Thus, melatonin has an oncostatic effect on some human tumors in vitro occasionally at physiological concentrations. PMID- 11022420 TI - [Monitoring of basic parameters in cancer patients]. PMID- 11022421 TI - [Time-dependent effects of antihypertensive agents and chronocorrecting action of melatonin in patients with arterial hypertension]. AB - The chronopharmacodynamics of angiotension-converting enzyme (ACE) inhibitors, such as ramipril (R), enalapril (E), acetene (A), caposide-50 (C-50) and the effect of the epiphyseal neurohormone melatonin used alone and in combination with cozaar (Co) and cintome (Ci) in the randomized groups including 124 patients with arterial hypertension (AH). The ACE inhibitors were given in the morning, afternoon, and evening. Co and Ci were used in the morning. Melatonin was given at 10.00 p.m. 10-14 days before and after therapy, 24-hour monitoring (Astracard, Russia) and echocardiography (Toshiba, Japan) were performed. Analyzing the findings indicated the time-dependent hemodynamic effects of R, E, and C-50. Ci, A, and Co were found to have antihypertensive and vasodilatory effects without normalizing the chronostructure of circadian rhythms of major hemodynamic parameters. By producing antihypertensive and vasodilatory effects, melatonin leads to the recovery of circadian hemodynamic organization when used alone and in combination with Co, Ci, and ACE inhibitors. PMID- 11022422 TI - [Spatial and temporary changes in sensitivity of murine proliferative system and energy exchange to typhoid fever infection]. AB - The authors studied the spatial and temporary organization of the proliferative system and energy exchange in the small intestinal epithelium, as well as spatial and temporary changes in the sensitivity of these systems in mice to typhoid fever infection (cultured Salmonella typhi, 4446) at day and night. The small intestinal epithelial systems were found to show a spatial and time organization and a close correlation between their temporary and spatial changes after infection. The temporary organization of the proliferative system in the esophageal epithelium is more sensitive to infection in the day-time and that of the small intestinal epithelial proliferative system is more marked at night. Thus, these changes are tissue-specific. Typhoid fever infection causes a negative effect on the proliferative system of the small intestine at night and on its energy exchange in the day-time, which shows the system-specific response of the temporary organization of a biological process within the same organ. The changes in the spatial proliferative and energy exchange parameters in the small intestinal epithelium, which have been caused by the infection of animals, depend on the time of a day. PMID- 11022423 TI - [Circadian rhythms of urinary parameters in children with glomerulonephritis]. AB - Circadian rhythms of urinary parameters were studied in healthy children and those with glomerulonephritis. In healthy children, urinary excretion of many substances rhythmically varied during a day. Variations were related to the volume and specific gravity of urine, to the levels of creatinine, MDA, and SOD activities. Children with glomerulonephritis showed changes in values of circadian rhythm of urine parameters. These disorders included changes in circadian variations and mean daily values, inversion of group and individual excretion patterns, which indicates that there is an external and internal disagreement of biological rhythms in children with glomerulonephritis. PMID- 11022424 TI - [Biocontrolled chronophysiotherapy for injuries and gastrointestinal diseases]. PMID- 11022425 TI - Improving the appropriateness of carotid endarterectomy: results of three city wide studies. PMID- 11022426 TI - Nurses must not be fooled by the pay award. PMID- 11022427 TI - National standards or pseudo-competencies? PMID- 11022428 TI - Case 19: nurse education. Nurse tutor who leaked examination answers to students. PMID- 11022429 TI - Management of long-term urethral catheters: minimizing complications. AB - Urinary tract infections, tissue damage and encrustation of the catheter, which may cause blockage, are all complications that can arise during long-term catheterization. It is important for nurses to provide effective catheter care in order to minimize the incidence of these complications. There is still controversy in the nursing literature about certain aspects of catheter management. This article explores a number of aspects of catheter care and suggests a rationale for effective and safe management, including the choice of catheter, choice of drainage system, care of the individual and the care of catheter and drainage system. PMID- 11022430 TI - Handwashing facilities in the clinical area: a literature review. AB - Handwashing is without doubt the most important intervention in the control of cross-infection. However, many healthcare staff do not comply with the procedure. One of the factors that contributes towards this is a lack of adequate and appropriate handwashing facilities. This article looks at the literature relating to handwashing and identifies what adequate and appropriate facilities actually are, with recommended standards for clinical areas. It concludes by highlighting that a greater commitment is needed from managers in this area in order to improve compliance with handwashing and therefore reduce infection rates. PMID- 11022431 TI - Improving standards for learning disabilities. PMID- 11022432 TI - A life story approach for people with profound learning disabilities. AB - This article presents an alternative approach to studying issues concerning people with profound learning disabilities. Previous approaches have relied too heavily on quantitative methods, e.g. questionnaires and rating scales, which demand a certain degree of verbal competence. When working with people with profound learning disabilities who can not talk, these techniques are clearly limited. The current research aims to redress this imbalance by presenting an in depth qualitative analysis of issues concerning the identities of people moving from a long-stay hospital to community care. Life story books were introduced as a resource for presenting biographical information about clients previously undocumented. It is suggested that we should move away from measuring to observing. Life story books take the focus away from viewing the participants as patients or subjects to considering them as people with their own unique life. PMID- 11022433 TI - Self-injurious behaviour: reviewing evidence for best practice. AB - This article brings together some of the research literature from the last 15 years concerning self-injurious behaviour (SIB) in people with learning disabilities. There is a focus on medical, biological and psychological research literature because of the important contribution these disciplines have made to the understanding of such behaviours. The article also explores contemporary evidence for the management of people with learning disabilities who present with SIB. The care and management of this group of people presents learning disability nurses with distinctively distressing challenges. Within the article definition, prevalence, topography, causation and evidence for best practice are outlined. The article concludes with a brief exploration of the role of the learning disability nurse in contributing to the health and wellbeing of this group of people. PMID- 11022434 TI - Gerontological nursing. 2: Refocusing the community agenda. AB - This, the second in a series of articles considering potential developments in gerontological nursing, focuses on a community setting. It highlights current trends in policy which emphasizes the promotion of independence and outlines implications for community nurses. The authors argue that if new roles are to emerge then the educational preparation of community nurses must reflect more fully future policy initiatives. PMID- 11022435 TI - Screening for hazardous drinking in a population of well women. AB - Previous research suggests that only about 15% of people who experience alcohol problems come to the notice of healthcare providers. Research has also shown that there is often a failure among healthcare providers to identify alcohol problems in patients whose primary presentation is not overtly alcohol related. It has been implied that this failure is contributed to by a lack of training and support on the part of the organization. This article reports on a project in which a group of health visitors were trained in a brief method of screening for and intervention with alcohol problems and then supported in identifying and intervening with women among their caseload who were drinking at unsafe levels. The sample comprised 430 'well women' between the ages of 18 and 45 years, with no known previous or current alcohol-related problem. All the women were on the caseloads of health visitors in one health board area and all women had children under the age of 16 years. It was found at screening that 13% of the sample consumed more than 14 units of alcohol per week. Consumption was measured 6 months after one session of minimal intervention, and it was found that 92% of the intervention group reduced their alcohol consumption. Mean consumption had decreased by 55%, from 21.11 units to 9.24 units per week. Among those women whose drinking had been at levels which were potentially problematic (capable of sustaining physical, psychological or social problems due to alcohol consumption), the mean weekly amount consumed decreased by 59% from 30.7 units weekly to 12.43 units. PMID- 11022436 TI - The use of Askina saline in the wound cleansing process. AB - After the removal of a dressing from a wound, wound cleansing is often the first action to be taken by the nurse. Saline or tap water is often used to loosen the dressing and therefore help to remove it. It can also be used for the subsequent cleansing of the wound if required. B/Braun Medical has a range of modern wound care products for all healing phases of acute and chronic wounds and this article will look at their saline range. PMID- 11022437 TI - Coming to terms with the death of a child. PMID- 11022438 TI - Web-based research: an innovative method for nursing research. AB - Although still in its infancy, the World Wide Web presents unparalleled opportunities for harnessing the power of the Internet to collect data from populations of interest in an economical and efficient manner. Web-based surveys offer a number of strengths including access to specific populations, speed of data access, and decreased data collection and data entry costs. These advantages are likely to increase as more people begin to use the Web as their primary means of communication. Nurse researchers who embrace this technology will become the pioneers in the new age of nursing research. PMID- 11022439 TI - The 1999 Schering Lecture. Cancer: the long and winding road. AB - Control, hope, uncertainty, and spirituality are not new phenomena, but perhaps we can see them in a different light. We still have much to learn about how these phenomena influence and are influenced by the cancer experience. I have asked you to reflect and consider how you, in your own practice, and your nursing colleagues can reframe cancer as a long-term condition. I have asked you to consider how this may change the ways in which you interact with clients and families. How refocusing might affect the care provided to travellers on the trip of a lifetime; the journey of cancer and the impact nursing can have on that journey. Are we ready to accept the opportunity to be part of someone's story--to journey with them? In their paper on the myth of control of suffering in palliative care, Gregory and English (1994) concluded that, "Reaching out is accomplished through presence, touch and careful listening" (p 22). People experiencing cancer choose those who will journey with them, offering a privilege of the maximal magnitude. PMID- 11022441 TI - Ashes to ashes: the significance of preventing lung cancer through primary prevention and health promotion. PMID- 11022440 TI - John's story: an application of the Calgary Family Intervention Model. AB - This article resulted from both authors sharing their professional experience; it illustrates an application of the Calgary Family Assessment Model (CFAM) and Calgary Family Intervention Model (CFIM) by Lorraine Wright and Maureen Leahey (1994). Indeed, some of the theoretical concepts presented by these authors were used to guide our analysis of the clinical situation. This brief intervention, consisting of four meetings, aimed to promote family coping during the palliative phase. Throughout each of the meetings, the nurse illustrates the principles guiding therapeutic conversations with the family, i.e. generating hypotheses, circularity, and neutrality. Through her intervention, she facilitates the continuation of the story, but without John. PMID- 11022442 TI - Consolidating the educational mandate for the CANO Pain Initiative. Canadian Association of Nurses in Oncology project group. PMID- 11022444 TI - 29th EDTNA/ERCA Conference. European Dialysis and Transplant Nurses Association European Renal Care Association. Lisbon, Portugal, 8-11 July 2000. Abstracts. PMID- 11022443 TI - Standardization of chemotherapy protocol summaries. PMID- 11022445 TI - Responding to the cues of suffering. AB - Enduring and suffering are fundamental and normal responses to catastrophic loss, yet strangely these two basic states have not been extensively explored using qualitative methods. Both states occur commonly as a human response to illness, injury, and bereavement, and it is the role of professional caregivers to respond to people who are in distress. In this article, I distinguish between states of enduring and suffering, discuss the distinctly different and appropriate comforting strategies to be used with each, and conclude by presenting questions for future research. PMID- 11022446 TI - Pregnancy, birth, and disability: women's health care experiences. AB - This qualitative study examined the pregnancy, birth and postpartum experiences of 12 women with mobility-limiting physical disabilities. Analysis of semi structured interviews of one to two hours revealed that the women's experiences were influenced by their own perspectives and the characteristics of health care system within which they were treated. The woman's experience included the effect of her disability, her resources, and her personality and approach. Health care system factors included provider attitudes, knowledge about disability and structural and political factors. Implications for health care providers are described. PMID- 11022447 TI - The role of social support and family relationships in women's responses to battering. AB - For women who are in abusive relationships, social support and close family relationships may be critical to the successful resolution of the abuse. Efforts to enhance battered women's support must be predicated on an understanding of women's perceptions of effective support and the constraints they experience to seeking support. In this paper we present findings from a qualitative analysis of semistructured interviews with 31 women who were in abusive relationships. The women were interviewed three times over two-and-a-half years. Results of the analysis included that women used female friends for support more often than family members, and that women were constrained from seeking support by (a) cultural and societal sanctions against leaving the relationship; (b) a pattern of caution in relating to others or forging new relationships; and (c) forced isolation/seeing self as isolative. Relationships with family members, especially parents, were not consistently seen as useful sources of support. PMID- 11022449 TI - Barriers to breast cancer screening: an integrative review. AB - Breast cancer has been increasing at an alarming rate and is considered to be of epidemic proportions in the United States, with current estimates indicating that 1 in 8 women will develop breast cancer during their lifetimes, according to Breast Cancer Facts and Figures, 1997, by the American Cancer Society [ACS]. In spite of the advances in technology to improve early diagnosis and an increased emphasis on education to promote awareness of early detection, 46,000 women die annually. A significant number of these losses could be prevented through risk reduction measures, yet many women do not practice breast self-exam (BSE) or receive adequate clinical screening. The purpose of this integrative review is to provide an analysis of the barriers to breast cancer screening with recommendations for future research. The studies will be categorized using the three modalities for breast cancer screening, mammography, clinical breast exam (CBE) and BSE. Demographic variables that impede breast cancer screening will be integrated into each section. The evidence is clear that in spite of breast cancer screening guidelines, increased awareness and access to mammography screening, women in the United States are not being screened adequately. Recommendations for future research will be included. PMID- 11022448 TI - Race differences in the relationships between dietary nutrients and overweight in women. AB - Increasingly overweight is a health problem in the United States, especially for African American (AA) women. Height, weight, and 24-hour dietary recall measurements from 46 European American (EA) and 44 AA women were used to assess the relationships between nutrients and overweight and the possible modifying effects of race. Significant race interactions were found when assessing the relationships between overweight and fats and mineral supplements. More EA women were overweight who had excessive fat intake, whereas more AA women were overweight who had appropriate fat intake. This reversed pattern for AA women was unexpected. Another unexpected finding was that overweight EA women used mineral supplements more frequently than nonoverweight EA women. These results were discussed in terms of energy imbalances, metabolic differences, and cultural differences. Additional studies of overweight assessing race and other biological factors that also include measures of energy expenditure and more precise measures of dietary energy intake are recommended. PMID- 11022451 TI - Development of a disease management program. AB - Changes in reimbursement, new regulatory guidelines, and other issues impacting the healthcare industry have forced providers to examine new models of healthcare delivery. One of those models is disease management or disease-state management. This article presents issues involved in developing such a program using the experiences of a Florida-based health system as an example. PMID- 11022450 TI - Women's experience of traumatic stress in cancer treatment. AB - In this study we investigated gender differences in the prevalence and predictors of posttraumatic stress disorder (PTSD) in a sample of cancer patients. Eighty seven patients (59 women and 28 men) recruited from a local cancer clinic completed measures of individual coping style, social support resources, symptoms of PTSD, and the nature of the trauma associated with these symptoms. PTSD symptoms were more frequently reported by women (27% versus 10%). Predictors of PTSD for women were (1) perceived intensity of cancer treatment; (2) problems with health care professionals; (3) and cognitive avoidant coping style. For men, the sole predictor of elevated PTSD scores was behavioral avoidance. Women reported significantly higher levels of treatment intensity (TI), greater numbers and types of treatment, and more problems with health care professionals. Interpersonal and relational aspects of their illness were reported as most stressful, compared with men who were more concerned with work and finances. These differences may, in part, reflect an androcentric model of cancer treatment, which women experience as particularly stressful. PMID- 11022452 TI - Management skills A to Z. PMID- 11022453 TI - A manager's wish list for 2000. PMID- 11022454 TI - Physician-based care teams. PMID- 11022455 TI - Regulation 2000 and beyond. PMID- 11022456 TI - Transformational leadership and personal values for managers in the new millennium. AB - Transformational leadership is best suited to the many demands that home health managers face daily. Transformational leaders who are driven by values that remain constant are better able to guide their agency. They are the ones able to make sound decisions now and have a vision for the future. PMID- 11022457 TI - The virtual home care office is here! PMID- 11022458 TI - Case management as a survival tool. PMID- 11022460 TI - Where do we go from here? PMID- 11022461 TI - [Staphylococcal necrotizing fasciitis: diagnosing and treating a hidden killer]. PMID- 11022459 TI - The importance of job satisfaction in retaining nurses. AB - This study's findings show how power and job satisfaction are perceived by home health nurses. It has important implications for managers and can be used in recruiting nurses. PMID- 11022462 TI - [Dyspnea crisis experienced by couples in their home]. PMID- 11022463 TI - [Antibiotic resistance: a preoccupying phenomenon]. PMID- 11022464 TI - [Traumatic amputation. Organization of first aid at the work place]. PMID- 11022465 TI - [Actions to be taken to assure the safety of the patient]. PMID- 11022466 TI - [The presence of a significant person and the "Sundown syndrome"]. PMID- 11022467 TI - A cognitive developmental approach to understanding how children cope with disasters. AB - TOPIC: This paper applies cognitive developmental theory to explain how preschoolers, school-age children, and adolescents process and respond to disasters. PURPOSE: To help clinicians understand the disaster experience from a child's point of view, recognize age-specific reactions, identify symptoms that may signal coping difficulties, and plan effective interventions. SOURCES: Case examples from the author's work with flood victims illustrate typical reactions for children (preschool to adolescents) within a family context, along with developmentally appropriate interventions. CONCLUSIONS: Children affected by disasters need nursing interventions geared toward their particular developmental level and sensitive to their perception of the disaster. PMID- 11022468 TI - Trauma group psychotherapy for the adolescent female client. AB - TOPIC: Therapeutic uses of adolescent trauma group psychotherapy for the treatment of post-traumatic stress disorder. PURPOSE: To demonstrate the role of the psychiatric nurse/group facilitator to prevent and treat adolescent emotional distress resulting from trauma. SOURCES: Published literature, author's clinical experience. CONCLUSIONS: Adolescent trauma group therapy is useful to deal with emotional, behavioral, maturational/developmental, physiological, cognitive, and solution-focused issues. PMID- 11022469 TI - Peer rejection and neglect of latency-age children: pathways and a group psychotherapy model. AB - TOPIC: Children's peer relations are one of the most powerful predictors of mental health PROBLEMS: Peer rejection and neglect are associated with depression, academic problems, school withdrawal, and delinquency. PURPOSE: To provide brief introductions to models of social development and peer rejection, and to introduce a developmental-group psychotherapy model for children experiencing peer neglect or rejection. SOURCES: Theoretical and empirical literature, as well as clinical experience. CONCLUSIONS: Early identification and intervention with children vulnerable to peer rejection are essential. Research addressing affective experiences of peer-rejected children and the relationships between self-perception and treatment outcomes is needed. PMID- 11022470 TI - A pregnant adolescent: negotiating a difficult journey to adulthood. PMID- 11022471 TI - Seizing the leadership moment. PMID- 11022472 TI - Request for feedback. PMID- 11022473 TI - Academic mentoring. Ensuring our future. PMID- 11022474 TI - Case management for children and families. PMID- 11022475 TI - Toilet training methods, clinical interventions, and recommendations. PMID- 11022476 TI - Physical abuse, social support, self-care, and pregnancy outcomes of older adolescents. PMID- 11022477 TI - Psychiatric comorbidity among adolescents with substance use disorders: findings from the MECA study. PMID- 11022478 TI - Do typical clinical doses of methylphenidate cause tics in children treated for attention-deficit hyperactivity disorder? PMID- 11022479 TI - Screening for major depression disorders in adolescent medical outpatients with the Beck Depression Inventory for Primary Care. PMID- 11022480 TI - Role clarity, organizational commitment, and job satisfaction during hospital reengineering. PMID- 11022481 TI - Pediatric telephone triage protocols: standardized decision-making or a false sense of security? PMID- 11022482 TI - Preparing presentations that produce peace of mind. PMID- 11022484 TI - Child and family nursing international consulting. The Web way. PMID- 11022483 TI - Diaries as a method of data collection with children. PMID- 11022485 TI - A specified outcome. PMID- 11022486 TI - Society for Adolescent Medicine Position Statement. Improving the nutritional health of adolescents. PMID- 11022487 TI - Conceptual analysis of compliance. AB - Compliance has been studied from a wide range of scientific perspectives including medicine, nursing, psychology and health economics. There is no agreement regarding a commonly accepted definition. Lack of consistency in the definition and measurement of compliance is a major problem in research which becomes more complicated in an international study. The response to the confusion over the term compliance has been to suggest and use alternative terms such as adherence, co-operation, mutuality and therapeutic alliance. These terms are ill defined and often are used as synonyms. The purpose of this paper is to analyse definitions of the concept of compliance. Abstracts from MEDLINE have been analysed in order to identify the types of compliance research that have been carried out. PMID- 11022488 TI - Health effects of work schedules in healthcare professions. AB - Increasing variety in working patterns and the appearance of new forms of shift schedules in the different occupational sectors, including health services, have raised great concern about the quality of working life and job performance. The aim of this paper is to present a systematic review of the literature on the effects on health of irregular schedules in healthcare professionals. Computer and manual searches of databases, and discussion with experts, were used to identify relevant studies. No conclusive evidence was found to favour any particular work system, although there is evidence that extended workdays (9-12 h) should be avoided as much as possible. There is need for carefully designed studies in order to evaluate the long-term consequences of work schedules in healthcare workers. PMID- 11022489 TI - Critical care nurses' decision-making activities in the natural clinical setting. AB - This article reports on observation of 18 nurses in urban and rural based critical care settings. The purpose of the study was to observe and describe the decision-making activities of critical care nurses within natural clinical settings. During the 2-hour observation, the researcher dictated a detailed commentary on to audio-tape of each nurse's actions. Tapes were transcribed and subjected to content analysis. Findings indicated three main categories of decisions. Decision frequencies were linked to nurses' critical care experience, appointment level, and location, as well as nursing shifts. The findings are discussed in relation to previous empirical evidence and the implications for practice. The author concludes that future research should be directed towards measuring the contextual influences on nurses' decision-making on the outcome of patient care. PMID- 11022490 TI - Information point: nursing career structure. PMID- 11022491 TI - Patients with implantable cardioverter-defibrillators and their conceptions of the life situation: a qualitative analysis. AB - The implantable cardioverter-defibrillator (ICD) is today widely used for the treatment of sudden cardiac near-death episodes as a result of malignant ventricular dysrhythmia. After examining the literature, only four descriptive studies, all carried out in the USA, with a qualitative analysis based on ICD patients' own perspectives on their life situation have been found. The aim of this study was to describe how patients living with an ICD-device in south western Sweden conceive their life situation. As the focus was on patients' conceptions seen from a holistic perspective, an analysis inspired by phenomenography was employed on a strategic sample of 15 ICD-patients. Six categories emerged: a feeling of safety, a feeling of gratitude, a feeling of being, having a network, having a belief in the future, and gaining awareness. Although the findings cannot be generalized because of the descriptive research design, they illuminate the beneficial as well as intrusive effects of such a device, and emphasize the need for support groups for patients and families as well as further education for personnel in hospital and primary health care. PMID- 11022492 TI - Needs of families with a relative in a critical care unit in Hong Kong. AB - The aim of this study is to explore family members' perceptions of their immediate needs following admission of a relative to a critical care unit in Hong Kong. A convenience sample of 30 family members was drawn from those available during the first 96 hours of hospitalization of their relative. Self-reported questionnaires, consisting of a demographic data sheet, a modified Chinese version of the 45-item Critical Care Family Needs Inventory (CCFNI) and semistructured interviews, are the instruments used to examine family members' perceptions of need importance and to ascertain whether or not these needs are met. Doctors and nurses are identified as the most suitable people to meet most immediate family needs. Conclusions are drawn as to the best focus of nursing interventions in order to provide quality care to patients and families. PMID- 11022493 TI - Assessment and prevention of shivering in patients with severe cerebral injury. A pilot study. AB - Low body temperature is induced by surface cooling to reduce metabolic demands in patients with severe cerebral injury. Shivering, which increases energy expenditure, is a common effect of surface cooling. The aim of this pilot study was to investigate whether increased gradient between the set point and peripheral temperature is related to shivering and whether modifying the loss of body heat during surface cooling decreases the frequency of shivering. Seven cerebrally injured patients at a neurosurgical ICU were studied. Shivering, surface cooling and the temperature gradient measured as the tympanic and the tip toe temperatures were registered every 30 min. Shivering was assessed at three levels: no shivering, mild shivering and severe shivering. In four patients the arms and legs were covered with a cotton cloth for part of the observation time to modify the rate of heat loss. The temperature measurements were repeated in each patient between 13 and 42 times. Four patients out of seven shivered. There was a significant association between increased temperature gradient and shivering (P < 0.01). Modifying the rate of heat loss decreased the temperature gradient (P < 0.001). By simultaneously measuring the tympanic and tip toe temperatures it may be possible to detect shivering earlier and decrease its frequency by modifying the loss of body heat during surface cooling. PMID- 11022494 TI - Waiting for a liver transplant: the experience of patients with familial amyloidotic polyneuropathy. AB - Liver transplant is a new treatment for familial amyloidotic polyneuropathy. The purpose of this phenomenological study is to describe the experience of waiting for a liver transplant from the familial amyloidotic polyneuropathy patients' perspective. Unstructured and open-ended interviews were conducted with 14 familial amyloidotic polyneuropathy patients and the analysis of data was inspired by Colaizzi's method. Waiting was found to involve two theme categories: waiting for a decision; and waiting for the operation. Seven themes were identified: bargaining with oneself; no influence/powerlessness; relief and joy; impatience; agony; time to prepare; and need for information and support. Implications for nursing practice, such as informational and emotional support, are discussed. PMID- 11022495 TI - The image of the children's nurse: a study of the qualities required by families of children's nurses' uniform. AB - Traditional views of the nurse see 'her' in a nurse's uniform, whether she be a 'sex symbol', 'angel of mercy' or 'battle axe'. Children's nursing has, over recent years, moved away from the idea of the uniform into more child friendly clothes, though the traditional uniform is still presented to the public by the media. The staff members of a Children's Outpatients Department (COPD) of a large acute hospital requested knowledge of the qualities families required from the uniform of nurses caring for them, prior to any change in uniform. This study aimed to develop an understanding of these qualities via the use of a survey presented to those attending the COPD. The findings indicate that there is a complex relationship between the qualities that families wish the uniform to present. A balance is needed between presenting a professional authority figure, providing security for families and suggesting an approachable figure, to allow effective communication between staff and families. PMID- 11022496 TI - Patient satisfaction with emergency nurse practitioners in A & E. AB - In recent years the complex process of managing services for patients requiring emergency care has been increasingly under scrutiny and considerable diversity has arisen in the organization of emergency care in both hospital and community settings. One innovation which has been particularly widespread is the development of Emergency Nurse Practitioners (ENPs): experienced Accident and Emergency nurses who assume entire responsibility for patients with minor injuries, in emergency settings. The present paper reports on a study which compared patients' satisfaction with care given by Emergency Nurse Practitioners with that provided by doctors and nurses working together in a traditional A & E. Patients were significantly more likely to have received health education and first aid advice from an ENP than from a doctor. They were also significantly more likely to have been given written instructions to take home and told whom to contact if they needed more help and advice following discharge. Those seen by an ENP also reported that they were subsequently significantly less worried about their health, than did patients seen in a traditional A & E. PMID- 11022498 TI - Infection control in wound care: a study of fatalism in community nursing. AB - As part of a study of community nurses' perceptions of quality in nursing care, the author conducted in-depth qualitative interviews with seven community-based nurses. As part of the study, nurses were asked to describe episodes of wound care and to discuss the factors which could affect the quality of such care. One of the most interesting themes to emerge from the data was the apparent ambivalence of the nurses' attitudes towards infection control in wound care. Nurses discussed the concept of 'aseptic technique' in fatalistic terms and seemed uncertain about what could be achieved in terms of infection control. Although their policy guidelines referred to 'aseptic technique', their educational experience appeared to have made them feel uncertain about the implementation of the measures involved. With the proviso that this was a small scale qualitative study, the author concludes by suggesting that there is a need for greater clarity, both in what is taught and in what is included in practice policy with regard to infection control in wound care. PMID- 11022497 TI - Improving hand hygiene in community healthcare settings: the impact of research and clinical collaboration. AB - Observation of nursing activities and bacteriological studies undertaken with a sample of nurses employed in a community NHS trust indicated that considerable scope for cross infection existed during domiciliary visits. Poor conditions in patients' homes compromised nurses' ability to perform hand hygiene effectively, increasing risks. A clinical trial indicated that carriage of medically significant bacteria likely to contribute to cross infection could be reduced by applying an antiseptic cream which exhibited residual effectiveness. An audit of hand hygiene throughout the inner city trust indicated the need to pay greater attention to hand hygiene, especially during home nursing visits. The situation was less acute in a rural trust where a second audit was performed for comparative purposes. The motivation of clinical staff to improve hand hygiene precautions was high. PMID- 11022499 TI - Strategies used in post-operative pain assessment and their clinical accuracy. AB - Our knowledge about the content of strategies used by staff members in a surgical recovery unit for assessment of post-operative pain is fairly limited. The aim of the present study was to describe variations in the content of strategies used by nurses and physicians in practical clinical pain assessments and to evaluate the clinical accuracy of the strategies used. Critical care nurses (n = 30), physicians (n = 30) and postsurgical patients (n = 180) comprise the respondents. Applying a phenomenographical approach, interview data were tape-recorded during 180 clinical pain assessments. The pain assessments were related to comparative bedside pain ratings (Visual analogue Scale, VAS), both by staff members and post operative patients. The recorded interviews were analysed to describe variations in ways of assessing pain. Pain assessment strategies were established by combining categories describing the impact of experience and categories of assessment criteria. The present observations, if included in the education of clinical staff members, could increase the understanding and thereby the quality of the pain assessment process. PMID- 11022500 TI - The effect of an acute pain service on nurses' knowledge and beliefs about post operative pain. AB - The management of post-operative pain has been an area of concern for many years, with many studies focusing on the knowledge and beliefs of nurses working in this area. Following the report of the Royal College of Surgeons & College of Anaesthetists (1990) in the UK, there has been a rapid expansion in the development of Acute Pain Services (APS) in an attempt to counter these concerns. This descriptive study considers the possible impact the introduction of an APS had on the knowledge and beliefs of nurses working in the surgical area. A closed answer questionnaire was used to replicate an earlier study (Mackintosh, 1994) which took place before the introduction of the APS. Findings demonstrate a consistent but mainly statistically non-significant trend in all areas towards an improved knowledge base and more appropriate beliefs about pain. PMID- 11022501 TI - Promoting autonomy and independence for older people within nursing practice: an observational study. AB - Although autonomy and independence are widely held to be important goals of care for older people, these terms lack precise definition and operationalization in the literature. This paper reports on observational data gathered as part of an evaluation of educational programmes in the nursing care of older people. Strategies which appear both to promote and inhibit autonomy and independence of older patients in a range of care settings are described. Recommendations are made for further research to explore the extent to which autonomy and independence are appropriate goals of care for older people with cognitive impairment. PMID- 11022502 TI - Reflections on intuition and expertise. AB - Reflective practice now appears firmly established in the English speaking world of professional nursing practice and development. Outside this linguistic context, however, the concept seems less well-known. This paper describes an experience drawn from clinical practice and education in French-speaking Switzerland followed by explicit reflection grounded in questions generated by Johns' model for structured reflection. Thus, a concept well-described in the English-language literature underpins an innovative approach to a French-language clinical teaching situation. The professional implications of this situation are explored through meaningful reflection providing new insight into familiar circumstances as they relate to the nurse tutor's role. This exploration is followed by a critical approach to the experience and the subsequent structured reflection in order to address relationships between intuition and expertise and self-awareness through reflection. A hermeneutic perspective provides additional insight into the nurse-patient relationship where both come to the situation with their own 'pre-understandings'. Individual horizons thus endorse a new understanding going beyond taken-for-granted meanings. PMID- 11022503 TI - A study of the effectiveness of different measuring times and counting methods of human radial pulse rates. AB - The purpose of this study was to determine the reliability and validity of measuring resting radial pulse rates by the use of three measuring times: 15, 30 and 60 s; and two counting methods: one beginning with zero (0) and the other with one (1). A two-factor within-subjects experimental design was used to determine the mean difference between pulse rates obtained from the radial artery, and the heart rates recorded by simultaneous electrocardiographic (ECG) recordings. The sample comprised 206 students. Mean difference was used to calculate the extent of any differences between radial pulse rates and the rate shown by the ECG. The interaction between measuring time and counting methods was confirmed using a two-factor within-subjects analysis of variance. For all types of measuring time, the counting from zero method produced a greater mean difference than the counting from one method. For all measuring times, the mean difference between radial pulse rates and rates shown by the ECG were non significant in the counting from one method. In other words, when the pulse rate is counted from one, the rates obtained at 15 or 30 s could be used to predict the one-minute resting pulse rates. The results of this study can contribute to the evidence base for this commonly used aspect of patient care. PMID- 11022504 TI - 'Emergency' consultation in general practice: patients' experiences. PMID- 11022505 TI - Qualitative and quantitative approaches in mental health nursing: moving the debate forward. AB - In recent years, two distinct, and apparently opposing, positions have arisen in debates concerned with mental health nursing research, education and practice. Each of the two has its champions and 'gurus', and its detractors. Writers associated with the quantitative camp have argued in favour of: experimental research where the aims are the investigation of the outcomes of nursing care; multidisciplinary rather than unidisciplinary research; a physiological explanation for mental illness; clinical practice which is evidence based, and skills based education. Qualitative camp writers, on the other hand, have argued for: qualitative research to explore the experiences of illness, and of nursing; the preservation of nursing-only research; the relative irrelevance of physical explanations of mental illness, and holistic nursing practice in which clinical techniques are less important than interpersonal relationships. In this paper, we first review and criticize some key ideas associated with these two positions. In our subsequent analysis, we argue that, rather than differing at some fundamental level, the differences found between the two camps may be more a matter of emphasis. The possibilities of reconciliation and synthesis are, finally, presented and discussed. PMID- 11022506 TI - Stress and burnout in community mental health nursing: a review of the literature. AB - There is a growing body of evidence that suggests that many community mental health nurses (CMHNs) experience considerable stress and burnout. This review aimed to bring together the research evidence in this area for CMHNs working within the UK. Seventeen papers were identified in the literature, seven of which looked at stress and burnout for all members of community mental health teams (CMHTs) and the remaining 10 papers focused on CMHNs. The evidence indicates that those health professionals working as part of community teams are experiencing increasing levels of stress and burnout as a result of increasing workloads, increasing administration and lack of resources. For CMHNs specific stressors were identified. These included increases in workload and administration, time management, inappropriate referrals, safety issues, role conflict, role ambiguity, lack of supervision, not having enough time for personal study and NHS reforms, general working conditions and lack of funding and resources. Areas for future research are described and the current study of Welsh CMHNs is announced. This review has been completed against a background of further significant changes in the health service. In the mental health field, specific new initiatives will have a significant impact on the practice of community mental health nursing. A new National Framework for Mental Health, along with a review of the Mental Health Act (1983), will undoubtedly help to shape the future practice of mental health nursing. PMID- 11022507 TI - Employment, social inclusion and mental health. AB - Whereas unemployment is clearly linked to mental health problems, employment can improve quality of life, mental health, social networks and social inclusion. Yet in the UK only 15% of people with serious mental health problems are employed- despite an overwhelming consensus from surveys, case studies and personal accounts that users want to work. This paper aims to challenge common misconceptions surrounding employment, work and mental health problems. Drawing on a range of research evidence and legislative guidance it discusses significant barriers to work and proposes feasible solutions. The need for mental health staff and services to become involved in the provision of work opportunities is considered, as is the vital role they can play in changing communities. The potency of work as a vehicle for improving the social inclusion and community tenure of people with mental health problems is highlighted. PMID- 11022508 TI - Dissociative identity disorder: a literature review. AB - This paper presents a review of the literature into dissociative identity disorder. This disorder, previously known as multiple personality disorder, is increasingly diagnosed, in part because of more focused diagnostic tools, but also because people are accessing services to assist with the longterm problems of early child abuse and neglect. Dissociative identity disorder is examined in the literature according to a variety of discourses, each of which suggest different ways of conceptualizing problems and therapeutic approaches. These discourses reviewed include: psychiatry, psychology, corporeality, feminism, social constructivism, anthropology, and postmodernism. The paper concludes with an examination of the nursing literature and suggests opportunities for nursing research into this complex mental health problem. PMID- 11022509 TI - Community, culture and character: the place of the virtues in psychiatric nursing practice. AB - In this paper, the authors address the serious neglect of the 'classical' virtues in ethical reflection upon mental health nursing. The virtues are offered as a possible alternative paradigm for mental health nursing in its search for new models and approaches. Embodied in the notion of a moral community, the virtues have important implications in addressing problems adherent in various 'dualisms' so described. By invoking the concepts of practical wisdom and praxis, the interrelationship between theory and practice, action and reflection and self and community, is shown. The contextual nature of the virtues is shown in a number of examples and their potential for the transformation of practice is shown. The opportunity is now present for mental health nursing to strengthen its basis for practice by distancing itself from scientific models. In so doing, it can embrace an approach which is consensual and contextual and which places ethical reflection at the centre of practice. PMID- 11022510 TI - A review of violence and personal injury legal cases in psychiatric and mental health nursing to identify a practical framework for risk assessment. AB - Legal cases were analyzed that involved violence resulting in staff injury, patient assaults on staff, accidents involving the elderly, and patient self harm or injury where there was also an alleged breech in the duty of care. The study resulted in recommendations for: improved systems of work, including a practical framework to anticipate and recognize risk, with a case management approach to individual care, and supervisory procedures for vulnerable staff, explicit consent from patients and carers, and integrated recording of interdisciplinary methods for care. PMID- 11022511 TI - Mental health service user involvement in nurse education: exploring the issues. AB - This paper reports on findings and issues arising from a study designed to promote mental health service users' involvement in a preregistration nursing curriculum. Users' views about the knowledge, skills and attributes required by mental health nurses were explored to inform the curriculum design. Strategies that would facilitate long term, active user involvement in the design and delivery of the curriculum were also explored. Findings are presented with concurrent discussion of issues arising from the research process in relation to user involvement in education. The issue of 'conflict' explores findings relating to users' views of a 'good' mental health nurse and inherent conflicts between user and professional views are highlighted. The representativeness of the research participants is explored and debated in relation to service user involvement in nurse education. Finally, the concepts of 'involvement' and 'tokenism' are discussed and recommendations made about how active user involvement in nurse education can be achieved. PMID- 11022512 TI - Anti oppressive practice: a route to the empowerment of people with dementia through communication and choice. AB - This short study looks at the issue of anti oppressive practice and the way that it relates to the care of people with dementia. In particular it considers ways in which people can be empowered and given choices around aspects of day-to-day living in a care setting. The study took place over a period of 6 months and utilized the Dementia Care Mapping (DCM) observation process to assess the level of well being of residents in the setting studied. The results can only be seen as a first stage in the process of understanding ways in which people with dementia may be empowered. It is demonstrated that a change of practice developed over the 6 months between the two observation periods. Areas such as communications and day-to-day activities are examined to identify both problems and strategies for such change. The conclusion highlights the need to continue the mapping exercise as a way of ensuring that change is not only maintained but also advanced. PMID- 11022513 TI - Constructing normality: a discourse analysis of the DSM-IV. AB - The purpose of this research was to explore how the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) 1994, (American Psychiatric Association, 1994) defines mental disorder and the theoretical assumptions upon which this is based. The analysis examines how the current definition has been constructed and what the criteria for specific mental disorders suggest about what is regarded as normal. The method employed for the research was a critical discourse analysis. This critical approach to research is primarily concerned with analysis of the use of language and the reproduction of dominant belief systems in discourse. It involves systematic and repeated readings of the DSM-IV (1994) to examine what evidence was employed by the text to substantiate its definition of mental disorder and how in the process some assumptions are made about what constitutes normality. This study challenges a central assumption in the DSM-IV's (1994) definition: that it is a pattern or syndrome 'that occurs in an individual'. The proposal that it occurs in an individual implies that it is a consequence of faulty individual functioning. This effectively excludes the social and cultural context in which experiences occur and ignores the role of discourse in shaping subjectivity and social relations. This study proposes that the definition and criteria for mental disorder are based on assumptions about normal behaviour that relate to productivity, unity, moderation and rationality. The influence of this authoritative image of normality pervades many areas of social life and pathologies experiences that could be regarded as responses to life events. PMID- 11022514 TI - 'I don't want to be a psychiatric nurse': an exploration of factors inhibiting recruitment to psychiatric nursing in Ireland. AB - Ireland has experienced a decline in recruitment to preregistration psychiatric nursing programmes (An Bord Altranais 1999). Social care workers often do similar psychosocial work with similar client groups served by psychiatric nurses. In marked contrast to psychiatric nursing, Irish social care diplomate and degree programmes are generally over subscribed. Yet graduates working in social care often experience inferiority in terms of pay, conditions and career structure compared to their psychiatric nursing counterparts (McElwee 1998). The question that therefore needs to be asked is why school leavers opt for such courses in preference to psychiatric nursing. This article describes the results of a pilot study, utilizing a focus group approach, to examine reasons for course and career choice among school leavers, psychiatric nursing students and social care students. Results indicate that school students rely on stereotypical views as part of their decisionmaking process in shaping occupational decisions. School students tend to conceptualize psychiatric nursing as being a job involving menial and physical tasks. Social care students saw psychiatric nursing as lacking autonomy and to be institutional in nature. Psychiatric nursing students themselves felt exploited and 'second class' compared to general nurses. Current recruitment campaigns and careers guidance within schools need to be more targeted on differentiating psychiatric nursing from its general nursing counterpart. PMID- 11022515 TI - Advanced nursing practice: experience, education and something else. PMID- 11022516 TI - The natural history, current status, and future trends of HIV infection. AB - Nearly 50 million people worldwide have been infected with human immunodeficiency virus (HIV), and an estimated 12 to 13 million children have been orphaned by the acquired immune deficiency syndrome (AIDS) epidemic. The natural history of HIV infection continues to evolve as researchers and clinicians unearth new facts about the virus and develop new treatment regimens for patients. The status of anti-HIV treatments is never static but constantly changing. Patients and providers struggle with adherence issues. Vaccine development, viewed as an essential step in controlling the epidemic, is complicated by the genetic diversity of the virus and the inability of the body to clear the virus. Tremendous strides have occurred in the HIV/AIDS arena, yet daunting challenges remain. This overview article presents the natural history, the current status, and future trends of HIV infection. PMID- 11022517 TI - Opportunistic infections in the era of highly active antiretroviral therapy. PMID- 11022518 TI - HIV counseling, testing, and prevention in primary care. AB - Human immunodeficiency virus (HIV) continues to be the second leading cause of death for persons ages 25 to 44 years in the United States, whereas new HIV infection rates remain steady. Coupled with the advent of new antiviral therapies that have significantly decreased mortality and morbidity rates, the importance of the primary care clinician in HIV prevention, early detection, and treatment is paramount. This article presents HIV risk analysis and prevention strategies for the primary care clinician practice. New HIV testing methods are reviewed as well as the Centers for Disease Control (CDC) pretest and posttest counseling guidelines. PMID- 11022520 TI - HIV/AIDS and women. AB - Women are the fastest growing segment of people with acquired immunodeficiency syndrome (AIDS), yet they often receive this diagnosis when the disease is in its advanced stages. New therapies have caused human immunodeficiency virus (HIV) to become a chronic and treatable disease for many of those afflicted. Primary care providers must be cognizant of the initial symptoms to facilitate early diagnosis and prompt treatment for women with HIV. Early signs of HIV in women are subtle. Providers must consider a diagnosis of HIV in women who present with vaginal infections, abnormal pap smears, or sexually transmitted diseases that are unusually severe, recurrent, and resistant to treatment efforts. These signs and symptoms, along with a corroborating history, may be early clues to HIV. Primary care providers, in conjunction with HIV specialists, must strive to decrease the incidence, morbidity, and mortality of the disease in women. PMID- 11022519 TI - Primary care for HIV-exposed and infected children: translating progress into practice. AB - Recent advances have changed the guidelines for diagnosing and managing pediatric human immunodeficiency virus (HIV) infection. HIV-exposed and HIV-infected children should be evaluated by, or in consultation with, pediatric HIV specialists. Primary care practitioners play a vital role in identification of infants and children at risk for HIV infection and can work collaboratively with pediatric HIV specialists to provide state-of-the-art care. With the use of perinatal zidovudine, perinatal transmission rates have been reduced to 3% to 4%, and they may be reduced even further by the use of combination antiretroviral therapy during pregnancy, viral load monitoring, and obstetric interventions. Diagnosis of HIV infection can be determined in all perinatally infected infants by 6 months of age. Combination antiretroviral therapy is the standard of care for HIV-infected children. It has become increasingly effective, but complex. Families living with HIV are affected by a number of psychosocial issues. Disclosure of HIV diagnosis to a child is an important clinical issue. As HIV infected children grow older, medical and psychosocial issues may impact school performance. The plan of care to address specific needs of HIV-infected children should be a collaborative effort between the children, their families, the primary care team, and the multidisciplinary pediatric HIV specialty team. PMID- 11022521 TI - Mental health and psychosocial issues in HIV care. AB - Human immunodeficiency virus (HIV) disease is associated with significant psychological stressors that require attention in their own right and can impact medical treatment. Psychological integration of illness occurs in identifiable stages and affects the patient's response to treatment interventions. Syndromes of depression, anxiety, stress, and substance abuse associated with HIV disease require recognition and appropriate treatment. The psychosocial context of treatment is also an important factor in HIV care, especially for special populations. Counseling and support groups are important tools in comprehensive HIV care and should be tailored to the needs of HIV/acquired immune deficiency syndrome (AIDS) patients. Mental health issues also exist for professionals working in HIV care. PMID- 11022522 TI - Antiretroviral treatment in HIV infection. PMID- 11022523 TI - The initial evaluation and management of an HIV(+) patient. PMID- 11022524 TI - Health care worker exposure. PMID- 11022525 TI - Patient education. HIV diagnostic testing. PMID- 11022526 TI - Patient education. HIV medications. PMID- 11022527 TI - Patient education. Nutrition and HIV. PMID- 11022528 TI - Young woman with recurrent yeast infections. PMID- 11022529 TI - Ownership and values: nurses, nursing and higher education. PMID- 11022530 TI - Modelling nursing activities: electronic patient records and their discontents. AB - A fully integrated and operating EPR in a clinical setting is hard to find: most applications can be found in outpatient or general practice settings or in isolated hospital wards. In clinical work practice problems with the electronic patient record (EPR) are frequent. These problems are at least partially due to the models of health care work embedded in EPRs. In this paper we will argue that these problems are at least partially due to the models of health care work embedded in current EPRs. We suggest that these models often contain projections of nurses' and doctors' work as it should be performed on the ward, rather than depicting how work is actually performed. We draw upon sociological insights to elucidate the fluid and pragmatic nature of healthcare work and give recommendations for the development of an empirically based EPR, which can support the work of nurses and other health care providers. We argue that these issues are of great importance to the nursing profession, since the EPR will help define the worksettings of the future. Since it is a tool that will impact the development of the nursing profession, nurses have and should have a stake in its development. PMID- 11022531 TI - Defining relationships and limiting power: two leaders of Australian nursing, 1868-1904. AB - This paper analyses aspects of the relationship between nursing and medicine during 1868-1904, in terms of power, gender and authority. A biographical approach is used with a focus on two leading nurses in Australia and their relationship with two leading medical practitioners. The first nurse is Lucy Osburn, the figurehead of the first generation of Nightingale nursing in Australia. The second nurse represents the second generation when Nightingale nursing had largely won acceptance and was firmly established in Australian hospitals: she is Susan McGahey. Their main medical antagonists were Dr Alfred Roberts and Dr Anderson Stuart. A struggle over the control of nursing is evident in these relationships. The outcome transcended personalities, greatly influenced the structure of modern nursing, and marked the rising tide of medical domination in Australia. PMID- 11022532 TI - Poststructuralism and nursing: uncomfortable bedfellows? AB - The benefits and limitations of the application of poststructuralist in nursing research are discussed. The debate concerning the use of poststructuralist theory in feminist research is drawn on to argue a divergence between a deconstructionist poststructuralism and nursing aims. It is argued that there are strong parallels between nursing and social movements such as feminism. The reasons why many feminist and nursing researchers have been attracted to poststructuralist theory are explored, as are the criticisms of poststructuralism as an inadequate, even reactionary theory. It is argued that poststructuralism is useful for examining or deconstructing existing discourse and practice, but incompatible and inadequate for research which aims to develop or change such discourses or practices. PMID- 11022533 TI - In the name of Husserl: nursing in pursuit of the things-in-themselves. AB - A perceived contradiction between the tenets of humanism and positivism secures phenomenology's endorsement in nursing as an alternative methodology to the natural sciences. Nursing's humanistic doctrine of valuing the individual is aligned with phenomenology in the belief that both projects investigate the subjective experiences of others. However, the belief that phenomenology opposes objectifying methods does not account for the different understandings of subjectivity that underpin various philosophic positions, such as humanism, phenomenology, hermeneutics and positivism. What is not recognised in nursing inquiry is that the same characteristics which belong to humanism and are subsequently attributed as belonging to phenomenology are grounded, philosophically, in a view that separates the person from her world. But rather than examining the premises on which phenomenology and humanism are based, nurse phenomenologists have simply yoked them together in a mutual alliance against a common enemy--Cartesian dualism. This paper focuses on Husserl and argues that the conceptual link between nursing as a humanistic discipline and Husserlian phenomenology cannot be sustained on philosophic and methodologic grounds. Far from being mutual allies, the philosophic underpinnings of humanism and Husserlian phenomenology offer two incommensurable views on the nature of subjectivity. PMID- 11022534 TI - Feeling old: being in a phase of transition in later life. AB - The aim of the study was to illuminate very old persons' experiences of feeling old in order to get a nuanced understanding of the ageing process in later life. Fifteen persons 85-96 years of age, living in their own homes, were interviewed in-depth. Data were analysed utilising a phenomenological-hermeneutic approach. Eight persons reported that they felt old. The experience of feeling old entailed four characteristics: being able to date the beginning of feeling old, fear of helplessness and of being unable to manage one's life situation, not recognising one's former self, and feeling different from others. These characteristics corresponded to the main properties of a transition process. The comprehensive understanding was that the very old persons who also felt old were in a phase of transition. The distinguishing qualities of those that felt old were also found to be in line with the dystonic dominance in the ninth stage designated by Joan Erikson of the previously described life cycle. Knowledge about transition processes and sensitivity to very old people's experiences of feeling old are of great importance to nursing in order to provide adequate health care services and prevent unhealthy transitions. PMID- 11022535 TI - Possibilities for critical social theory and Foucault's work: a toolbox approach. AB - The benefits and constraints of philosophical frameworks using the work of Michel Foucault and critical social theorists, such as Fay, Giroux and McLaren, are examined in the light of their traditions. The reasons nurse researchers adopt these frameworks are explored, as are the tensions between the respective theories. A complementary 'toolbox' approach to the research process addresses some of the theoretical and methodological challenges presented by each framework. Such an approach provides distinctive insights into nursing practice that the other has ignored or missed. It is argued that by converging the two frameworks into a toolbox approach, it is possible to examine or deconstruct existing practices, whilst also providing an avenue for nurses to reconstruct or change such practices. PMID- 11022536 TI - Clinical exchange: one model to achieve culturally sensitive care. AB - This paper reports on a clinical exchange programme that formed part of a pre registration European nursing degree run by three collaborating institutions in England, Holland and Spain. The course included: common and shared learning including two summer schools; and the development of a second language before the students went on a three-month clinical placement in one of the other base institutions' clinical environments. The aim of the course was to enable students to become culturally sensitive carers. This was achieved by developing a programme based on transcultural nursing principles in theory and practice. Data were gathered by interview, focus groups, and questionnaires from 79 exchange students, fostering the strategies of illuminative evaluation. The paper examines: how the aims of the course were met; the factors that inhibited the attainment of certain goals; and how the acquisition of a second language influenced the students' learning about nursing. A model is presented to illustrate the process of transformative learning from the exchange experience. PMID- 11022538 TI - Setting a trap for hantavirus. PMID- 11022537 TI - Ending the blame game. PMID- 11022539 TI - Applying a wrist restraint. PMID- 11022540 TI - Klebsiella pneumoniae. PMID- 11022541 TI - Using pulse oximetry. PMID- 11022542 TI - Action stat. Blow-out fracture. PMID- 11022543 TI - What's your gut reaction to a poison emergency? PMID- 11022545 TI - My privilege. PMID- 11022544 TI - Skin tears: staying on guard against the enemy of frail skin. PMID- 11022546 TI - Asking the right questions about medication safety. PMID- 11022547 TI - Looking at depression through bifocal lenses. PMID- 11022548 TI - Managing complications of i.v. therapy. PMID- 11022549 TI - Sending the wrong signals. PMID- 11022550 TI - Myths & facts...about organ donation. PMID- 11022551 TI - Safe computer charting. PMID- 11022552 TI - The question. PMID- 11022553 TI - Olanzapine and rhabdomyolysis. PMID- 11022554 TI - How to use gauze dressings. PMID- 11022555 TI - Spotlight on correctional facilities nursing. PMID- 11022556 TI - Don't argue with the arbitrator. PMID- 11022557 TI - Reaching out. PMID- 11022558 TI - [All of us are responsible for a human care for the aged]. PMID- 11022559 TI - [Importance of quality management]. PMID- 11022560 TI - [Mail-order drugs for diabetics. A comparison is important]. PMID- 11022561 TI - [Need to assure more safety for female patients and their nurses in an old age home]. PMID- 11022562 TI - [The seven pillars of wound treatment]. PMID- 11022563 TI - [A new interdisciplinary project]. PMID- 11022564 TI - [The economic importance of nursing services can be present in health policy]. PMID- 11022566 TI - [Quality control and assurance in nursing care: need for changes]. PMID- 11022565 TI - [Improved management of decubitus ulcer is advantageous to the patient as well as to the economy]. PMID- 11022567 TI - [Lack of justification for therapy or surgery: need for facilitating patient information]. PMID- 11022568 TI - [Verbalizing feelings (emotions)]. PMID- 11022569 TI - [Ethical aspects of health care economy]. AB - One of the present-day problems which illustrates several ethical issues is the development of health care, generally referred to as health policy. For over 25 years there has been a debate relating to runaway costs in health care. This led to various retrenchment attempts which failed altogether and will probably continue to fail in the future unless there is a radical change in approaching the subject. Brief explanations regarding the economies of public health care will show that by means of this science statements concerning the consequences as regards costs and efficiency can be made, but the material organisation of a health care system remains somewhat vague. The same applies to ethics as a science. To be able to explain how a health care system can be organised, one has to take a stand based on the system of values relevant to the particular individual. Yet the question is not, whether a certain concept is right or good, but rather to what extent it is plausible. It can be illustrated that the case against restricting health services cannot be justified. Following this there will be a discussion as to who is actually authorized to enforce such restrictions in a society with competing value systems, and which criteria can be applied. Finally the author will try to conceptualize the situation, taking a stand based upon his own views and beliefs. PMID- 11022570 TI - [Demands on interpersonal and communicative skills of nurses in their daily practice]. AB - This article explores the objectives for developing interpersonal skills in nursing education proceeding from the demands of daily nursing practice. The typical situations and structures of communication between nurses and patients are ascertained by a qualitative study. It comes to the conclusion that this communication pattern is strongly characterized by power structures and that nurses often miss out on a differentiated and adequate view of these real requirements and conditions. From the results of the study conclusions for nursing education are drawn. Furthermore, the guiding principles for communication skills in nursing that correspond with a client-centered direction of interaction in order to give psychological help are discussed and rejected. PMID- 11022571 TI - [Balance of power in hospitals]. AB - It is known that there is a large extent of working dissatisfaction within some professional groups of the health service system. Especially in the hospital sector, many "struggles for power" take place. Unfortunately, these struggles are often only examined with regard to individual points of view without considering the system-oriented background. The following text will reflect the discussion in social sciences as to the distribution of power within the health service system. The double meaning of the distribution of power will be explained and, by means of decisive phenomenons, the development of cost and efficiency structures on the one hand as well as the development of health political processes in decision making on the other hand will be described. In the section "The Hospital in the 20th Century", the structures that still can be widely found nowadays will be specified and examined as to aspects in behavioural sciences. Finally, as a logical consequence, an urgently needed reorientation from administration towards management will be pointed out. PMID- 11022572 TI - [Social interaction between nurses and female patients in the course oforganization of a Nursing Development Unit]. AB - Following Friedson's work in the early 1970s, discussion about nursing's status as a profession took place. As a result the nursing development unit movement started in Great Britain in the early 1980ies. The goal of nursing development units is committed to both the development of nurses and nursing practice. The programme was supported by a grant of 3.2 million pound sterling from the British Department of Health. The last decade has witnessed a proliferation of these units. There a now over 300 (Draper, 1996). In Germany this programme of nursing development units is still rather unknown. The author believes that it should be discussed also in Germany as an effective instrument for the development of nursing as both an academic and a practice discipline. The central focus was to draw attention to the social interactions between nurses and patients within an NDU. Another question was whether nursing development units fundamentally represent an effective way to improve the standard of patient care by promoting excellence in nursing. In conclusion the level of patient oriented care and the quality of social interactions between nurses and patients has increased. However, examples for backward steps in the development of the unit the author visited were also found. The study shows that there were both conditions which inhibited and which gave support nurses' development and their practice. PMID- 11022573 TI - [Visiting regulation in intensive care units. Review of English-language literature 1984-98]. AB - This review analyses the Anglo-American nursing literature of the past 15 years that focuses on the subject of visiting policies at intensive care units. It is the aim of this paper to prepare a scientific base for discussion on ICU visiting policies in German nursing literature. Part 1 describes three visiting models whose concepts can be clearly differentiated. This is followed by the argumentation of the described models. Part 2 puts the focus on studies that have investigated visiting needs of patients as well as studies, in which the effect of visits and visiting models on patients has been researched. Transferability of results to the situation in Germany seems to be difficult for two reasons. First, all studies took part in a different cultural context. Secondly, most studies are limited by small size and convenient samples. Nevertheless, some studies indicate trends and support each other through similar findings. While reviewing the literature, it turned out that there has been an intensive debate about ICU visiting policies in Anglo-American nursing journals. This debate can not be held without involving and questioning nurses' professional understanding of patient care. Requirements for nurses as a result of possible changes of the visiting policies are presented and discussed. The article ends with suggestions for future research in Germany. PMID- 11022574 TI - [Nursing and education in gerontology]. AB - Originally, nursing was a practically determined concept. Modern nursing theories are scientifically based concepts, which--more or less--are open to psychosocial aspects. As to seniors, educational gerontology favours nursing theories, which focus upon psychosocial components. Such nursing theories are important prerequisites for successful cooperation between nursing and educational gerontology. PMID- 11022575 TI - [Michael Fischer (1887-1948)--life and work of an important founder of Catholic nursing in Germany]. AB - In 1919, Michael Fischer OSC was appointed to the German charity union "Zentrale des Deutschen Caritasverbandes" (DCV) in Freiburg. He assumed his duties as the acting manager and general secretary for the German catholic union of health institutions "Verband Katholischer Kranken- und Pflegeanstalten". For nearly twenty years, he was involved in expanding and strengthening this specialized organisation. For more than a decade, Michael Fischer influenced catholic medical care in Germany by holding lectures; on the whole, he published the medical welfare journal "Krankendienst" as well as fifteen specialized books and more than five hundred articles in different journals, which were essential. For all his efforts, his enormous engagement, historical health research has ignored him until now. The following essay is meant to give insight into his life and works and pays tribute to his importance in nursing care. PMID- 11022576 TI - The effects of linking research (evidence-based practice) to policy development. PMID- 11022577 TI - Nurse in profile. Daniel Prentice. PMID- 11022578 TI - Time for a new Canada Health Act. PMID- 11022579 TI - Update from the Canadian Dyspepsia Working Group. PMID- 11022580 TI - A parent and a doctor. PMID- 11022581 TI - Battling opiate overdoses. PMID- 11022582 TI - Attitudinal problems facing international medical graduates. PMID- 11022583 TI - Be careful how you report survey results. PMID- 11022584 TI - Comparison of human papillomavirus DNA testing and repeat Papanicolaou test in women with low-grade cervical cytologic abnormalities: a randomized trial. HPV Effectiveness in Lowgrade Paps (HELP) Study No. 1 Group. AB - BACKGROUND: Results of cervical cytology screening showing atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesions (LSIL) indicate risk for high-grade cervical intraepithelial neoplasia (CIN 2 or 3). In a community-based randomized trial we compared the test performance of human papillomavirus (HPV) DNA testing with that of 6-month repeat Papanicolaou (Pap) test in detecting histologically confirmed CIN 2 or 3. METHODS: We randomly assigned 212 women aged 16-50 years with ASCUS or LSIL on cervical cytology screening to undergo either immediate HPV DNA testing or a repeat Pap test in 6 months. Cervical swabs for the HPV DNA testing and the Pap smears were obtained by their family physicians. We tested the swabs for oncogenic HPV using the Hybrid Capture II assay (Digene Corp., Beltsville, Md.). Community-based pathologists examined the Pap smears. All women were referred for colposcopy by their family physicians. Two gynecological pathologists assessed the histology findings. We calculated test performance in women who completed the trial using CIN 2 or 3 as the reference standard. RESULTS: A total of 159 women completed the study. Compared with HPV DNA testing, which detected 87.5% (7/8) of the cases of CIN 2 or 3, repeat Pap smear showing high-grade intraepithelial neoplasia (HSIL) detected 11.1% (1/9) of cases (p = 0.004), and repeat Pap smear showing ASCUS, LSIL or HSIL detected 55.6% (5/9) (p = 0.16). Corresponding specificities were 50.6%, 95.2% (p = 0.002) and 55.6% (p = 0.61). Loss to follow up was 17.1% in the HPV test group and 32.7% in the repeat Pap group (p = 0.009). Given the 7 cases of CIN 2 or 3 detected by HPV testing and the 5 cases detected by the repeat Pap smear, the incremental cost of HPV testing was calculated to be $3003 per additional case of CIN identified. INTERPRETATION: HPV DNA testing was more costly but was associated with significantly less loss to follow-up. It may detect more cases of CIN 2 or 3 in women with low-grade cytologic abnormalities. PMID- 11022586 TI - Incidence of major depression in Canada. PMID- 11022585 TI - Does the site of postgraduate family medicine training predict performance on summative examinations? A comparison of urban and remote programs. AB - BACKGROUND: The location of postgraduate medical training is shifting from teaching hospitals in urban centres to community practice in rural and remote settings. We were interested in knowing whether learning, as measured by summative examinations, was comparable between graduates who trained in urban centres and those who trained in remote and rural settings. METHODS: Family medicine training programs in Ontario were selected as a model of postgraduate medical training. The results of the 2 summative examinations--the Medical Council of Canada Qualifying Examination (MCCQE) Part II and the College of Family Physicians of Canada (CFPC) certification examination--for graduates of the programs at Ontario's 5 medical schools were compared with the results for graduates of the programs in Sudbury and Thunder Bay from 1994 to 1997. The comparability of these 2 cohorts at entry into training was evaluated using the results of their MCCQE Part I, completed just before the family medicine training. RESULTS: Between 1994 and 1997, 1013 graduates of family medicine programs (922 at the medical schools and 91 at the remote sites) completed the CFPC certification examination; a subset of 663 completed both the MCCQE Part I and the MCCQE Part II. The MCCQE Part I results for graduates in the remote programs did not differ significantly from those for graduates entering the programs in the medical schools (mean score 531.3 [standard deviation (SD) 69.8] and 521.8 [SD 74.4] respectively, p = 0.33). The MCCQE Part II results did not differ significantly between the 2 groups either (mean score 555.1 [SD 71.7] and 545.0 [SD 76.4] respectively, p = 0.32). Similarly, there were no consistent, significant differences in the results of the CFPC certification examination between the 2 groups. INTERPRETATION: In this model of postgraduate medical training, learning was comparable between trainees in urban family medicine programs and those in rural, community-based programs. The reasons why this outcome might be unexpected and the limitations on the generalizability of these results are discussed. PMID- 11022587 TI - Who wrote this paper anyway?The new Vancouver Group statement refines the definition of authorship. PMID- 11022588 TI - Rheumatology: 8. Advanced therapy. PMID- 11022589 TI - Environment and health: 2. Global climate change and health. PMID- 11022590 TI - The wellness program for medical faculty at the University of Ottawa: a work in progress. PMID- 11022592 TI - Montrealers ante up for private surgery. PMID- 11022591 TI - Tomato lycopene and its role in human health and chronic diseases. AB - Lycopene is a carotenoid that is present in tomatoes, processed tomato products and other fruits. It is one of the most potent antioxidants among dietary carotenoids. Dietary intake of tomatoes and tomato products containing lycopene has been shown to be associated with a decreased risk of chronic diseases, such as cancer and cardiovascular disease. Serum and tissue lycopene levels have been found to be inversely related to the incidence of several types of cancer, including breast cancer and prostate cancer. Although the antioxidant properties of lycopene are thought to be primarily responsible for its beneficial effects, evidence is accumulating to suggest that other mechanisms may also be involved. In this article we outline the possible mechanisms of action of lycopene and review the current understanding of its role in human health and disease prevention. PMID- 11022593 TI - A clinical-decision rule for cervical spine injury. PMID- 11022594 TI - Use of a critical pathway for the management of community-acquired pneumonia: the CAPITAL Study. PMID- 11022595 TI - Tornadoes and disaster management. PMID- 11022596 TI - The future of medicine. PMID- 11022597 TI - [DMW special topic issue: Gastroenterology]. PMID- 11022598 TI - [Benign tumors of the Vater's papilla]. AB - BACKGROUND AND OBJECTIVE: Villous adenoma is the most common tumour of the papilla of Vater, and transition from adenoma to carcinoma is now generally accepted as proven. It is thus essential for an adenoma to be removed. Methods of removal have ranged from endoscopic sling papillectomy to partial duodenopancreatectomy. It was the aim of this study to determine whether recurrence can be prevented by local resection. PATIENTS AND METHODS: 58 patients with a benign tumour of the papilla (26 men, 32 women; average age 59 [range from 18 to 81] years) were included. Depending on preoperative histology, intraoperative frozen-section diagnosis and macroscopic histopathology, some form of surgical intervention was undertaken, most often resection of the ampulla. The clinical course and findings at postoperative follow-up were recorded and the absence of recurrence checked by endoscopy. RESULTS: Ampullectomy was performed in 49 of the 58 patients, papillectomy in three. Although frozen-section examination had failed to reveal any malignancy, resection of the head of the pancreas with preservation of the pylorus was done in six patients, carcinoma having been suspected macroscopically. There were no operative deaths. An adenoma had been found in 44 patients, one quarter of them showing severe dysplasia: follow-up examinations after a mean interval of 45 months (range of 6-180 months) failed to find any recurrence. CONCLUSION: Ampullectomy provides an adequate surgical treatment of benign adenoma of the ampulla of Vater. Accurate surgical technique and pre-, intra- and final histopathological diagnosis by an experienced pathologist are decisive factors in determining the ultimate outcome. If the histological findings as to benignity are unclear, resection of the head of the pancreas with preservation of the pylorus by an experienced surgeon is indicated. PMID- 11022599 TI - [Gastric polyps in pernicious anemia: indications for polypectomy]. AB - HISTORY AND CLINICAL FINDINGS: A 54-year-old man was admitted to hospital with dyspnoea, giddiness, lack of appetite and burning sensation in the tongue for 3 years. Gastritis was known from his history. Clinical examination showed jaundice, glossitis, systolic murmur and pallhypesthesia of both legs. INVESTIGATIONS: Blood and bone marrow investigations revealed pernicious anaemia. At gastroscopy, fundal gastritis was diagnosed and two small polyps of the fundus were removed. Histologically, one of the two polyps turned out to be a gastric carcinoid type 1 (according to [16]). Additional investigations (x-ray, ultrasound of the abdomen, somatostatin-receptor scintigraphy) did not find any metastases. TREATMENT AND COURSE: With the diagnosis of a pernicious anaemia the patient was treated with vitamin B12. The original symptoms and the anaemia disappeared. Additional therapy of the gastric carcinoid was not necessary. One year after beginning of the treatment, no signs of the gastric carcinoid were found. CONCLUSION: In gastric polyps of patients with pernicious anemia neoplastic transformations will often be found. That is why polypectomy should always be done. The best therapy of the gastric carcinoids type 1, which are the most frequently gastric carcinoids, is endoscopic tumour removal. Surgery is the treatment of choice only for gastric carcinoids type 2 to 4. Concerning quality of life and costs, endoscopic treatment is better for patients with gastric carcinoids type 1 than conventional surgical therapy. PMID- 11022600 TI - [Indications for liver biopsy in liver tumors]. PMID- 11022601 TI - [Colonic carcinoma associated with ulcerative colitis. Risk factors, molecular pathogenesis and surveillance]. PMID- 11022602 TI - [Conservative treatment of gallstones: pro]. PMID- 11022603 TI - [In Process Citation] PMID- 11022604 TI - [Concerning the debate on nosocomial transmission of hepatitis C by infected medical personnel]. PMID- 11022605 TI - [The 125th anniversary of the DMW (Deutsche Medizinische Wochenschrift): development and perspectives]. PMID- 11022607 TI - [Medical errors]. PMID- 11022606 TI - [Heparin after knee joint arthroscopy]. PMID- 11022608 TI - [Female pseudohermaphroditism in congenital adrenogenital syndrome as an incidental intraoperative finding in a 68-year-old patient]. PMID- 11022609 TI - Abuse of the assistant's fee as a 'kickback'. PMID- 11022610 TI - Prevention of vertical transmission of HIV--a constitutional catch-22 situation. PMID- 11022611 TI - HIV--the threat to South Africa's blood transfusion services. PMID- 11022612 TI - Trauma and alcohol. PMID- 11022613 TI - Aorto-iliac occlusive disease in the various population groups of South Africa. PMID- 11022614 TI - Anaesthesia in rural hospitals. PMID- 11022615 TI - Abortion: damned if you do or you don't. PMID- 11022616 TI - We can detect drug abuse--but will we? PMID- 11022617 TI - Get pragmatic about pot. PMID- 11022618 TI - Is flying unhealthy? PMID- 11022619 TI - Nobel Laureate Slams Pfizer. PMID- 11022620 TI - When to hire another doctor and make it pay. PMID- 11022621 TI - HIV vaccine trials in South Africa--some ethical considerations. PMID- 11022622 TI - The Employment Equity Act--another HIV calamity. PMID- 11022623 TI - Insulinoma presenting as temporal lobe epilepsy. PMID- 11022624 TI - Spinal epidural abscess. PMID- 11022625 TI - Legalizing advance directives in South Africa. PMID- 11022626 TI - Absence of commonly recurring BRCA1 mutations in black South African women with breast cancer. PMID- 11022627 TI - Shoe size and pelvic adequacy. PMID- 11022628 TI - Treatment of head injuries in the public sector in South Africa. PMID- 11022629 TI - National and provincial estimated costs and cost effectiveness of a programme to reduce mother-to-child HIV transmission in South Africa. AB - OBJECTIVE: To estimate the cost and cost effectiveness nationally and for each province of a programme to reduce mother-to-child transmission (MTCT) of HIV in South Africa. METHODS: A model developed to estimate cost and cost effectiveness of interventions in Hlabisa, KwaZulu-Natal, was modified and applied to each province. This model predicts a 37% reduction in paediatric HIV infections if short-course oral zidovudine (ZDV) plus infant formula feed for 4 months is provided within a strengthened health system. Estimates of the number of pregnancies and HIV prevalence among pregnant women per province in 1997 were combined with an estimated 30% MTCT rate. Costs were calculated from a health system perspective, and effectiveness was estimated as cost per infection averted and cost per disability-adjusted life year (DALY) gained. RESULTS: In 1997, 63,397 paediatric HIV infections were estimated to have occurred in South Africa, mainly in KwaZulu-Natal (18,513, 29%) and Gauteng (10,417, 16%). The cost of a national programme is estimated at R155.9 million (1997 rand costs, 0.94% of the national health budget). Major cost items are drugs (R46.4 m, 30%), staff salaries (R45.8 m, 29%), and formula feed (R37.1 m, 24%). Most money would need to be spent in KwaZulu-Natal (R37.6 m, 24% of national cost), Gauteng (R25.2 m, 16%) and the Eastern Cape (R24 m, 15%). National cost per infection averted is R6,724, and R213 per DALY gained. Provincial DALY costs range from R176 to R369. CONCLUSIONS: A national programme preventing 37% of expected paediatric HIV infections would cost a small fraction of the national health budget, at a cost equivalent to R3.89 per capita total population. The cost per DALY gained compares well with established public health and clinical interventions in middle income countries, even without factoring in the care costs that would be saved through a successful programme. Cost effectiveness is greatest where HIV prevalence is highest. PMID- 11022630 TI - From policy to praxis--a framework for the delivery of district mental health care in KwaZulu-Natal. AB - This article provides a schema for the provision of mental health care at district level. A framework for service delivery was derived from research conducted by the Community Mental Health Programme (CMHP) into the development of aspects of a district mental health care system in a semi-rural community area in KwaZulu-Natal. Furthermore, information was drawn from interviews with key stakeholders, national and provincial policy documents as well as international experience in the implementation of community-based systems of mental health care. PMID- 11022631 TI - The Cape Town experience with haematopoietic stem cell transplantation: the paediatric programme. AB - OBJECTIVE: To determine the outcome of children with blood malignancies and bone marrow failure syndromes treated by paediatricians in the context of an adult haematopoietic transplantation programme. DESIGN: Retrospective chart review. SETTING: Hospital wards in a provincial tertiary institution in the Western Cape (Department of Haematology, Groote Schuur Hospital). SUBJECTS: Twenty-eight hospitalised children with haematological malignancies (acute lymphoblastic leukaemia (ALL) N = 4, acute myeloblastic leukaemia (AML) N = 13), or bone marrow failure syndromes (N = 11), who consecutively received autologous or allogeneic marrow grafts from HLA-identical siblings. OUTCOME MEASURES: Children (younger than 18 years) received allogeneic or autologous stem cell transplants. In the former group, two forms of graft-versus-host disease (GVHD) prophylaxis were used. Conditioning with radiation-containing regimens was followed by stem cell product infusion after T-cell depletion (CAMPATH 1, ex vivo immunoglobulin G (IgG); rat anti CD52). Children with malignancies who received unfractionated grafts were myeloablated, mainly with busulfan 16 mg/kg and cyclophosphamide 120 mg/kg. Those affected by marrow failure were prepared with cyclophosphamide and antilymphocyte globulin. Median age at time of transplantation was 116 months (range 18-212 months). The main cause of death was disease recurrence (N = 5) and GVHD (N = 3). Twenty-one children survived, 11 of 16 in complete remission (CR) from malignancy. Nine of the eleven patients presenting with marrow failure and 1 patient with severe combined immunodeficiency (SCID) remained disease free at a median follow-up of 934 days (range 70-2,330 days). Significantly longer disease free (P = 0.03) and overall survival (P = 0.05, Cox Mantel test) was experienced by those who received T-cell-depleted stem cell grafts. CONCLUSIONS: The strategy of T-cell depletion of bone marrow/blood stem cells from HLA-matched siblings for transplantation into children with blood disorders has been successful and cost effective. These favourable results are the consequence of rational co-operation between adult and paediatric transplant physicians. PMID- 11022632 TI - An evaluation of the effectiveness of nutrition advisers in three rural areas of northern province. AB - OBJECTIVE: To evaluate the efficacy of a nutrition education intervention programme undertaken by trained local women (nutrition advisers) in rural villages in Northern Province. The programme was aimed at the caregivers of infants living in those villages. STUDY DESIGN: A cross-sectional survey design undertaken in 1989 and again in 1992. STUDY POPULATION: Female caregivers of infants living in three study villages (study area (SA)) and three control villages (control area (CA)) in Northern Province. Households were randomly selected. The response rate of households in the SA and CA was 70% (N = 1,040) at baseline and 84% (N = 1,263) after intervention. METHODS: A baseline study was undertaken in the SA and CA in 1989. A questionnaire eliciting sociodemographic data and information on dietary practices and nutritional knowledge was developed and used in the baseline study and after intervention. A nutrition education programme was undertaken by 6 trained local women (nutrition advisers) in the SA. After 2 years the survey was repeated using the same questionnaire in both the SA and CA. The SA and CA were compared with regard to breast-feeding and infant feeding practices; use of milk, brown bread, legumes and nuts; and use of school lunch boxes by older children. RESULTS: The percentage of women who initiated breast-feeding on the day of birth improved significantly in the SA from 60% to 90%. The frequency of feeding infants at 6 months improved significantly in the SA (P < 0.01). The introduction of solid foods to infants on the first day of life decreased from 26.5% to 6.3% in the SA. There were no significant differences in the SA only with regard to use of milk, brown bread, legumes, nuts, and school lunch boxes. However, some positive findings were a greater increase in the use of these items in the SA compared with the CA. CONCLUSION: A nutrition education programme undertaken by trained local women can significantly improve breast-feeding and infant feeding practices in rural areas. PMID- 11022633 TI - [Clinical course and diagnosis of the acute craniocervical injury]. AB - Combined observation and treatment of 126 patients (80 men, 46 women) aged 15-67 years were carried out in the acute period of craniocerebral trauma of mild or moderate severity. Diagnostic methods including neurological examination, manual testing and X-ray investigation are discussed. The conclusion is made that craniocerebral trauma should be considered as craniocervical trauma as an indirect damage of the neck was revealed in each case in the form of myofascial syndrome pronounced in different degree; cervical syndrome of vertebral artery was also frequent. These data must be taken into consideration in pathogenetic treatment of patients with craniocerebral trauma. PMID- 11022634 TI - [The role of brain stem in determination of compensation of mechanisms regulating autonomic functions as illustrated by craniocerebral trauma]. AB - Clinical neurologic and electroencepalographic examinations were performed in 484 patients with craniocerebral trauma (CCT) of different degree of severity and in remote period of severe CCT. Polygraphic study of autonomic functions including ECG, digital photoplethysmography and evaluation of dermal galvanic reactions were performed in 114 patients. A comparison of the disorders in autonomic functions both with a level and a degree of irritation of brain stem structures, according to EEG-data permitted to define a degree of impairment of compensatory mechanisms responsible for autonomic functions' regulation in CCT of different severity. PMID- 11022635 TI - [The influence of radiation of the child mental development]. AB - The paper presents the results of implementation of the first stage of WHO programme concerning consequences of the Chernobyl accident in terms of influence of radiation on mental health of children whose mothers were exposed to radiation. 4210 children born in the period from April 26, 1986 to February 1, 1987 in Byelorussia, Ukraine, Russia in contaminated and non-polluted (control) regions. In radiation-contaminated regions there was higher frequency of mental retardation and other mental disorders as well as deterioration of both mental and physic health of their parents. A preliminary character of the data obtained is emphasized as well as the necessity of more detailed follow-up of these groups. PMID- 11022636 TI - [Mental disorders in men enlisted into military service and in soldiers (clinico epidemiological study)]. AB - A total clinical epidemiologic survey was performed in one of Moscow regions in 1985-1990. On the first stage 2660 medical records of the individuals unfitted for military service were picked out. 42.8% of them had mental diseases. From 2102 medical records of the soldiers and sailors there were 66 of the discharged from the army. 45.5% of them were dismissed because of mental disease. On the second stage, 30 men dismissed because of mental disease and 299 men called for military service but discharged because of mental disease were examined. The duration of the follow-up was from 2 to 8 years. The most frequent reason both for the discharge from the army and for unfittness because of medical indications were borderline mental diseases (56.6% in the group of the discharged from the army and 40.9% in the group of men called up for military service). Endogenic diseases (schizophrenia, manic-depressive psychosis) made up only 16.6 and 15.8%, respectively. It was found that personal peculiarities and structure of mental diseases as well as risk factors of microsocial and biological nature were the most important indices for fitness for military service and for rehabilitative prophylactic work in soldiers during the first 6 months of the service. PMID- 11022637 TI - [Treatment of infantile cerebral paralysis using transcraniovertebral microelectric stimulation]. AB - A new method of therapy of children with cerebral palsy (CP) has been elaborated. Transitory consequent action of weak bipolar low-frequency current (30-100 mcA, 2 10 Hz) on different levels of motor activity regulation (brain and spinal cord, effectory organs) resulted in a stable correction of both motor and other types of disorders in CP patients. Therapeutic course included 2-8 procedures, 1-2 weekly, their duration 3-5 minutes. Depending on the reaction on the transcraniovertebral microelectrostimulation (MES) the patients were divided into fast and slow reactive groups. Different reaction of the children to MES reflected, probably, a variety of pathogenetic CP mechanisms and a mosaic of cerebral damage. Corrective MES action on the patients' brain was polysystemic and was, probably, based on the changes in integral brain mechanisms, voluminous transmission and homeostasis of brain, in particular. PMID- 11022638 TI - [Mental disorders in acute neuroleptic poisoning]. AB - 110 patients (52 men, 58 women) in a state of acute intoxication were examined in emergency department of psychoneurologic hospital. Neuroleptics of different groups were used to commit a suicide (91.3% of the cases), to obtain a toxicomanic effect (6.3%) and accidentally (1.8%). The patients suffered from borderline mental disorders (39.2%), schizophrenia (40.9%), manic-depressive psychosis in depressive phase (10.9%), chronic alcoholism (4.5%) and organic damages of CNS (4.5%). The patients with borderline states used various drugs and had more light disorders of consciousness (deafness). Meanwhile, the patients with endogenic mental disorders used strong neuroleptics, as a rule, that resulted in coma. In residual period there were different syndromes from asthenic to psychoorganic ones. They were more severe after poisoning with aminazinum, haloperidol, leponex. PMID- 11022639 TI - [EEG diagnostic potential in the estimation of the severity and sequelae of ischemic stroke]. AB - Simultaneous consideration of both electroencephalographic and neurologic data as well as the results of both computer and emission computer tomography in patients with consequences of ischemic stroke during a process of neurorehabilitation gave chance for more accurate definition of organic origin of some EEG phenomena. It was found that a damage of the conductive pathways between the cortex and subcortical structures associated with disorders of blood circulation in the area of the middle cerebral artery was accompanied by local disorders in electric activity of the cortex mainly in form of polymorphic waves of theta- and delta ranges and groups of theta-waves. Besides, a character of the slow waves depended on the size of the damage and on the degree of a decrease of blood circulation in the area of the damage. A decrease of the damage focus and improvement of circulation during restoration process was accompanied by changes in local deviations in cortical electric activity: polymorphic slow waves activity was replaced by sharp waves of alpha- and theta-ranges. Location of the changes of electric activity in caudal regions of the cortex is an unfavourable prognostic sign for restoration of motor activity in patients with ischemic stroke. PMID- 11022641 TI - [Immunopharmacological method for the diagnosis of multiple sclerosis]. AB - Using the data on increased level of opioids in the internal media of patients with multiple sclerosis it was found that blood lymphocytes from MS patients had elevated adhesive activity to opiates, including morphine, in the reaction of specific rosette formation. Immunopharmacologic method for MS diagnosis is proposed based on detection of considerably increased level of the rosette forming cells, specific to morphine (morphine test). This test is a reliable additional marker for all clinical forms of the disease and provides differential diagnosis of both MS and some other diseases with similar clinical symptomatology at any age, any duration and a character of a course of the disease. PMID- 11022640 TI - [Friedreich's disease: a real spectrum of clinical manifestations in terms of direct DNA diagnosis]. AB - Friedreich's disease (FD) is autosomal-recessive form of hereditary ataxias conditioned by expansion of the trinucleotide GAA-repetitions in a new X25 gene. The study was performed in 20 patients from 13 families of different ethnic origin (Slavs, Turkmen, Moldavians, etc) with a suspicion to FD as well as in their 24 relatives who were clinically healthy. Direct DNA-diagnosis confirmed FD diagnosis in patients from 11 families; besides, a number of GAA-repetitions in the patients was in the range from 100 till 1200 (680 +/- 350). A molecular analysis revealed that FD severity was determined by a character of the mutation in each case: a classic form of the disease was characterized by the highest degree of the expansion of GAA-repetitions (more than 400), meanwhile atypic FD cases with late debut and slow progression were conditioned by either a small degree of the expansion of GAA-repetitions or by the presence of point mutations in one of the gene's alleles. A direct DNA-diagnosis permitted to determine a heterozygous carriage of the mutation in 3 clinically healthy individuals. Such cases are necessary to take into consideration in medico-genetic consultations. PMID- 11022642 TI - [Pathogenetic mechanisms and clinical peculiarities of trigeminal facial pain]. AB - 316 patients with extracranial, intracranial and intracerebral lesions of the trigeminal system were examined. Together with a neurologic observation, the examination included registration of trigeminal somatosensory, acoustic brain stem and visual evoked potentials as well as electroencephalography (including brain mapping) and magnetic resonance tomography of the brain. Etiologic factors, location of the damage and systemic pathogenetic mechanisms characteristic for the main types of both paroxysmal and nonparoxysmal facial pain were established. Peculiarities of the manifestations of trigeminal neuralgia, neuropathy and differential facial pain are discussed. A comparative analysis of peculiarities of an algesic syndrome in three forms of verified radiculopathies conditioned by the lesion of trigeminal, glossopharyngeal and cerebrospinal nerves was performed. An original classification of trigeminal facial pain is proposed. PMID- 11022643 TI - [Seasonal incidence of alcoholic psychoses]. AB - Analysis of the incidence of alcoholic psychoses in Moscow in 1982-1993 revealed a considerable increase of their number during the period from March till July August and a decrease by October-November. It was found that such increase resulted from low-concentration atmospheric oxygen in spring-summer period. Hypoxia as a factor provoking alcoholic psychosis is responsible for 10-12% of the cases. The highest sensitivity to hypoxia occurred in the last days of the abuse. PMID- 11022644 TI - [Autonomic homeostasis in healthy men at different level of retrograde transport of Na+/Li+ in erythrocyte membrane]. PMID- 11022645 TI - [A rare case of "mirror" writing]. PMID- 11022646 TI - [Current knowledge of cochleovestibular dysfunctions]. PMID- 11022648 TI - [Evolution of the pharmacology of hypnotic drugs: from alcohol to zolpidem]. PMID- 11022647 TI - [Blood-brain barrier]. PMID- 11022649 TI - [Comment on the article by I.G. Morozova "Classification of pathological drives in children and adolescents"]. PMID- 11022650 TI - [Predictors of prognosis after angioplasty in the unprotected left main coronary artery]. AB - OBJECTIVES: The short-term and long-term predictors of outcome after coronary angioplasty in the unprotected left main coronary artery were investigated. METHODS: The procedure was performed in 122 consecutive patients for de novo lesions without myocardial infarction in our hospital between April 1986 and October 1998, including 16 emergency cases. Procedures were directional coronary atherectomy (73 patients), balloon angioplasty (31 patients), and stent implantation (18 patients). There were 101 males and mean age was 68 +/- 10 years. Follow-up angiography was performed in 98% of discharged patients, and all patients were clinically followed up for more than 1 year. Clinical and angiographic predictors of in-hospital and long-term outcome were evaluated. RESULTS: The in-hospital mortality was 5.7% (7 of 122 patients). Multivariate analysis revealed that more patients were admitted as emergency cases (57% vs 10%, p = 0.0088), with left ventricular ejection fraction < or = 35% (57% vs 22%, p = 0.029) and renal failure (43% vs 3%, p = 0.0004) finally died. Mean follow-up period was 3.5 years. Estimated survival rate was 77.1%, and cardiac-death free survival rate was 81.0% at 5 years by the Kaplan-Meier method. Univariate analysis showed that the predictors of cardiac death were emergency angioplasty, renal failure, decreased left ventricular ejection fraction, multivessel disease and unstable angina and/or congestive heart failure. Cox's regression model showed that renal failure (p = 0.0004) and multivessel disease (p = 0.0075) were significant predictors of long-term prognosis. CONCLUSIONS: Renal failure was the strongest predictor of outcome after unprotected left main coronary artery angioplasty. PMID- 11022651 TI - [Respiratory function after coronary artery bypass grafting through mini sternotomy as a factor of early recovery]. AB - OBJECTIVES: Early recovery in patients after minimally invasive coronary artery bypass grafting with mini-sternotomy and cardiopulmonary bypass (MICS-CABG) was compared to standard CABG by assessing preoperative and postoperative (7 to 10 days after) respiratory function. METHODS: Fifteen patients (Group M; mean age 62.1 years) underwent MICS-CABG with a mean of 2.3 distal anastomoses per patient. Ten patients (Group F; mean age 63.8 years) underwent standard CABG through full sternotomy with a mean of 2.4 distal anastomoses per patient. RESULTS: Postoperative coronary angiography showed that the patency rate of the grafts was 97% in Group M and 96% in Group F. Intubation time and hospital stay were significantly shorter (p < 0.01) in Group M (6.2 +/- 2.4 hours, 16.3 +/- 3.1 days) than in Group F (10.8 +/- 2.9 hours, 22.8 +/- 2.5 days). Respiratory function measured as the percentage of postoperative to preoperative values (Group M/Group F; mean +/- standard error) were vital capacity of 95.8 +/- 3.1%/74.6 +/- 3.4% (p < 0.05), 1 sec percentage of forced expiratory volume of 98.8 +/- 2.3%/71.8 +/- 2.8% (p < 0.05) and peak expiratory flow rate of 91.7 +/- 4.2%/89.4 +/- 4.5%. CONCLUSIONS: Quick recovery of the respiratory function after MICS-CABG may be important in the early recovery and short hospital stay of MICS CABG patients compared with standard CABG patients. PMID- 11022652 TI - [Increased incidence of coronary artery disease and cardiac death in elderly diabetic nephropathy patients undergoing chronic hemodialysis therapy]. AB - OBJECTIVES: The incidence of coronary artery disease and cardiac death was investigated in elderly diabetic patients undergoing chronic hemodialysis therapy. METHODS: Three hundred thirty-five patients who began hemodialysis therapy since 1992 were followed up by echocardiography and treadmill exercise testing. Coronary angiography was also performed in patients with angina pectoris. Angina pectoris was defined as clinical symptoms > Canadian Cardiovascular Society classification II, and asynergy findings by echocardiography or ST depression > 0.1 mV during the treadmill exercise test. Coronary artery stenosis was defined as narrowing > or = 75%. Patients were divided into 4 groups: diabetic nephropathy (DN) > or = 65 years old (Group O/DN, n = 56), DN < 65 years old (Group Y/DN, n = 84), non-DN > or = 65 years old (Group O/non-DN, n = 76) and non-DN < 65 years old (Group Y/non-DN, n = 119). RESULTS: Between 1992 and 1998, there were 137 patients with angina pectoris (40.9%), 79 with coronary artery stenosis (23.6%) and 37 with cardiac death (11.0%). Cumulative incidences of angina pectoris, coronary artery stenosis and cardiac death were significantly higher in the following order of groups; O/DN > Y/DN > O/non-DN > Y/non-DN. Five-year cumulative incidences of angina pectoris, coronary artery stenosis and cardiac death in Groups O/DN vs Y/non-DN were 72.2% vs 38.6%, 53.7% vs 12.2% and 50.6% vs 3.5%, respectively. Relative risks of aging and diabetic nephropathy for angina pectoris, coronary artery stenosis and cardiac death were 3.8, 7.9 and 22.4, respectively (p < 0.0001). CONCLUSIONS: Aging and the presence of diabetes are strong risk factors for coronary artery disease and cardiac death in hemodialysis patients. Therefore, diagnosis and treatment of coronary artery disease should be achieved at the early stage of hemodialysis therapy. PMID- 11022653 TI - [Anatomical variations of Koch's triangle in patients with atrioventricular nodal reentrant tachycardia: usefulness of the left anterior oblique view]. AB - OBJECTIVES: Several anatomical distances of Koch's triangle including the ablation site were measured and correlated with clinical features and slow pathway potentials in patients with atrioventricular nodal reentrant tachycardia to improve the avoidance of complete atrioventricular block. METHODS: Sixty consecutive patients (24 males and 36 females, mean age 47 +/- 12 years) with successfully eliminated atrioventricular nodal reentrat tachycardia were studied. The distances between the His-bundle area and the base of the coronary sinus ostium (Dis HBE-CS) and the distances between the successful ablation site and the base of the CS ostium (Dis SP-CS) were measured in both right anterior oblique and left anterior oblique views, and used to define the dimensions of Koch's triangle. The relationship between the slow pathway potentials at the successful ablation site and anatomical distances was estimated. RESULTS: The Dis HBE-CS in the right anterior oblique view was negatively correlated with patient age (r = -0.759, p < 0.001) and body mass index. In contrast, the Dis HBE-CS in the left anterior oblique view had only weak correlations with patient age and body mass index. The mechanism of the short Dis HBE-CS in the right anterior oblique view in elderly obese patients tended to change the shape of the tricuspid annulus from a circle to an ellipse, compressed by the ascending aorta and diaphragma. The Dis SP-CS in the right anterior oblique view associated with the low frequency potential (Haissaguerre's slow pathway potential) was longer than that associated with the high frequency potential (Jackman's slow pathway potential). CONCLUSIONS: Elderly obese patients had shorter distances between the proximal His-bundle area and the base of the coronary sinus ostium in the right anterior oblique view. In contrast, the Dis HBE-CS in the left anterior oblique view was not so narrow. Therefore, slow pathway ablation can be performed safely without complicated complete atrioventricular block, using both the slow pathway potential guided approach and the anatomical guided approach, especially in the left anterior oblique view. PMID- 11022654 TI - [Is myocardial impairment in idiopathic dilated hypertrophic cardiomyopathy reversible by carvedilol?: a case report]. AB - A 64-year-old woman with dilated hypertrophic cardiomyopathy was treated by beta blocker, because of recurrent episodes of congestive heart failure. Carvedilol administration was started with a dose of 2.5 mg/day, and gradually increased to the dose of 20 mg/day 3 months later. Her functional class recovered from New York Heart Association stage IV to stage II. Thallium-201 scintigraphy demonstrated advanced myocardial damage: defect/low perfusion in the anteroseptal, infero-posterior and apical regions. Iodine-123 beta-methyl-p iodophenyl-pentadecanoic acid scintigraphy confirmed these findings. However, the scintigraphic abnormalities had reversed to near normal perfusion and metabolism similar to those of stunned myocardium after an ischemic episode. This case demonstrates the recovery of myocardial impairment during beta-blocker therapy in a patient with dilated hypertrophic cardiomyopathy. PMID- 11022655 TI - [Recurrent pulmonary embolism from left subclavian thrombosis: a case report]. AB - A 38-year-old woman was admitted to our hospital because of pulmonary thromboembolism. Thrombolysis therapy resulted in initial improvement in symptoms and laboratory data. However, 4 months later, pulmonary thromboembolism recurred despite antiplatelet and anticoagulation therapy. Contrast venography and venous ultrasonography of both upper and lower extremities revealed subtotal occlusion and venous thrombosis of the left subclavian vein with collateral vessels, but no evidence of lower extremity venous thrombosis. She had no history of subclavian venous catheterization, neoplasm, hypercoagulability or other predisposing cause of thrombus formation. Operative ligation of the left subclavian vein was performed at the junction with the internal jugular vein. White thrombus was identified within the venous lumen. She was well without recurrent pulmonary thromboembolism or venous insufficiency for 10 months after the operation. Surgical interruption of the subclavian vein may be effective to prevent recurrent pulmonary thromboembolism in patients with recurrent pulmonary thromboembolism due to venous thrombosis of the upper extremity despite therapeutic anticoagulation. PMID- 11022656 TI - [A 69-year-old woman with congestive heart failure caused by myocardial ischemia and severe anemia]. PMID- 11022658 TI - [Opinions and repartees. Direct speech at the DOC Congress, Nuremberg]. PMID- 11022657 TI - [A 78-year-old man complaining of chest oppression at rest]. PMID- 11022659 TI - [Mid- and long-term results after trabeculectomy in patients with juvenile and late-juvenile open-angle glaucoma]. AB - BACKGROUND: To determine success rate of trabeculotomy in patients with juvenile and late-juvenile glaucoma in a retrospective study. PATIENTS AND METHODS: 46 eyes of 41 juvenile glaucoma-patients (24 female, 17 male) were submitted to trabeculotomy during the period of 1980-1997. Patients age at the time of surgery was 30.2 +/- 10.5 years (11-49 years, median 21 years). Follow-up time was 35 +/- 46 months (1-155 months). RESULTS: Median preoperative visual acuity was 1.0 (range 0.002 to 1.25), postoperative visual acuity at the end of the follow up period was 0.8 (median) with a range from 0.002 to 1.2. In all patients preoperative intraocular pressure was elevated and a glaucomatous configuration of the optic disc was noticed. Preoperative intraocular pressure (IOP) ranged from 30.5 +/- 13.2 mm Hg, after surgery IOP was 16.2 +/- 5 mm Hg. Mean decrease of the IOP postoperatively was 54.4%. In 37% of the eyes IOP was lower than 23 mm Hg without additional medication, in 47% of the eyes IOP was lower than 23 mm Hg with additional medication. In 8 eyes additional antiglaucoma-surgery was necessary. After a follow-up time of 60 months 81% of the patients (n = 37 eyes) had an IOP below 23 mm Hg, after 120 months in 80% of the patients (n = 29 eyes) IOP was below 23 mm Hg. CONCLUSION: The success rate of trabeculotomy (defined as the ability to lower intraocular pressure lower than 23 mm Hg with or without antiglaucoma medication) in our patients is 84%. After second operation, success rate is 89%. Trabeculotomy for patients with juvenile open angle glaucoma is a safe procedure and avoids problems and complications of filtering procedures. PMID- 11022660 TI - [Frequency and risk factors of intraocular pressure increase after penetrating keratoplasty]. AB - PURPOSE: The aim of this study was to evaluate the incidence, risk factors and management of glaucoma following penetrating keratoplasty (PK) and to check for possible correlations with the indication for PK. PATIENTS AND METHODS: 534 eyes of 483 patients (age 54.7 +/- 19.8 years), that had undergone PK from 1989 to 1999 were retrospectively analyzed. The mean follow-up was 2.7 +/- 1.8 years. According to the type of surgical procedure patients were classified in 5 groups: group 1) PK only (n = 291); group 2) PK combined with extracapsular cataract extraction and intraocular lens (IOL) implantation (n = 124); group 3) PK combined with secundary IOL-implantation or IOL-change (n = 32); group 4) PK only after previous cataract surgery (n = 55) and group 5) Cataract surgery after PK (n = 32). The trephination was performed from the epithelial side in donor and recipient either by nonmechanical trephination with the 193 nm excimer laser (n = 444) or mechanical trephination (n = 90). An iridotomy was performed routinely during PK. The postoperative treatment with topical steroids was standardized. RESULTS: Preoperatively, glaucoma was diagnosed in 3.2% for all patients with groups 3 und 4 showing a significantly larger number of glaucomatous eyes (15.6% and 18.9%). Most of the mild early intraocular pressure elevations in group 1 (3.4%) and group 2 (3.0%) were controlled after 6 months (0.5% and 1.1%). Groups 3 and 4 showed most frequently increased intraocular pressure and/or antiglaucomatous treatment (21.4% and 18.7%) six weeks postoperatively. Patients in all groups showed no higher prevalence of persistent glaucoma from six months postoperatively than preoperatively throughout the follow-up period. Again groups 3 and 4 presented the largest number of persistent glaucoma. In most eyes the IOP was controlled by topical antiglaucomatous treatment. Serious forms seen in 2 patients with anterior synechiae caused by anterior chamber lens required several filtering surgeries or pars plicata diathermy, respectively. One patient with pseudoexfoliation syndrome required laser trabeculoplasty. The so-called "Urrets Zavalia syndrome" was seen in none of the patients. CONCLUSION: The development of increased intraocular pressure after PK varied with the indication for keratoplasty. Postkeratoplasty glaucoma seems to be strongly associated with preexisting anterior synechiae and/or simultaneous anterior chamber lens implant removal. Intraoperative iridotomy can prevent the development of acute angle closure glaucoma after keratoplasty. PMID- 11022661 TI - [Monitoring of morphometric changes of optic discs with morphologic progression of glaucomatous optic atrophy by means of laser scanner tomography]. AB - AIM: Aim of this study was to measure morphometric changes in optic discs with laser-scanning tomography (HRT, Heidelberg-Retina-Tomograph, Heidelberg) in eyes with early glaucomatous morphologic progression. PATIENTS AND METHODS: 61 eyes of 36 patients with marked neuroretinal rim loss or its early morphologic signs (1. optic disc hemorrhages, 2. reduced visibility of the retinal nerve fiber layer (RNF), 3. appearance of narrowing of retinal vessels, 4. enlargement of the choroidal, parapapillary atrophy) were compared to 74 normal eyes of 39 probands. 15 degrees stereographs of the optic discs were evaluated for morphologic changes. The morphometric variables of the neuroretinal rim and excavation measured by the HRT were examined in the course of the disease. RESULTS: In the group of normals no significant changes of the neuroretinal rim in the course of 2.0 +/- 1.2 years were found. In the group of glaucomatous eyes (3.0 +/- 1.5 years follow-up) 34 eyes showed marked neuroretinal rim loss, 17 disc hemorrhages, 4 vessel narrowing, 3 an increased chorioidal atrophy, 3 a decreased visibility of the retinal nerve fiber layer. In these eyes a significant loss of rim area (p = 0.01) and an increase of excavation area (p = 0.0001) and volume (p = 0.003) was measured by the HRT. Only three eyes showed a perimetric loss of sensitivity (0.8-3.4 db) in Octopus static perimetry. CONCLUSIONS: Laser-scanning tomography of the optic disc seems to be able to measure morphometric changes in eyes with morphologic progression of glaucomatous optic atrophy, even before perimetric changes occur. PMID- 11022662 TI - [Utilization of a new night vision enhancement device (DAVIS)]. AB - PATIENTS AND METHODS: Following complete ophthalmologic examination 37 patients with night blindness due to Retinitis Pigmentosa (sometimes Usher-Syndrome) and Choroideremia (n = 3) performed several tests with DAVIS during darkness. We evaluated the improvement of visual function on a special outside course in the city of Heidelberg (duration 1.5 to 4 hours). RESULTS: Twenty six of the patients were able to better recognize obstacles, 28 could see objects which were not seen without DAVIS. Twenty two of the 37 patients would use the DAVIS. Patients needed a visual acuity of more than 0.1 and more than 6 degree of central visual field to experience improvement with DAVIS. However, in patients with only minimal changes of the visual field, the restriction due to the presence of the device was a drawback. Sudden occurrence of light sources leads to blinding and limits the indoor use. CONCLUSION: DAVIS enhances contrast acuity especially during night and twilight. This leads to improvement of orientation due to better recognition of obstacles and allows rehabilitation of patients with night blindness for outdoor mobility. Individual test and adjustment of DAVIS is necessary to allow exact and adequate prescription. PMID- 11022663 TI - [Choroidal changes after photodynamic therapy (PDT). A two-year follow-up study of 38 patients]. AB - BACKGROUND: Photodynamic therapy is a new option for treatment of choroidal neovascularisation in patients with age-related macular degeneration. But choroidal changes and associated angiographic characteristics have not been further evaluated. PATIENTS: Indocyanine green angiography was used to follow 38 patients with subfoveal choroidal neovascularisation in age-related macular degeneration over up to two years. All patients were treated with the photosensitizer Benzoporphyrin Derivative-MA receiving either a single or triple treatment. RESULTS: Indocyanine green angiography shows two effects of photodynamic therapy. On the one hand a selective and lasting closure of choroidal neovascularisation was documented. Choroidal neovascularisation-size and leakage was significantly reduced in the entire treatment group to 20.7% and 28.3% one week after treatment, followed by a slow increase to 33.3% and 41.2% at up to two years longterm follow up. On the other hand photodynamic therapy causes typically a peri-lesional hypofluorescence in Indocyanine green angiography. This hypofluorescence is most likely due to choroidal hypoperfusion and vascular endothelial changes. A continuous increase in fluorescence was shown, reaching again 90% of the pretreatment intensity at 3 months, documenting a good recovery of the choroidal network. CONCLUSION: The results show that photodynamic therapy is an alternative treatment in age-related macular degeneration with choroidal, subfoveal neovascularisation. Indocyaningreen angiography reflects well choroidal changes associated with this therapy and may be helpful to choose treatment intervals. PMID- 11022664 TI - [Unilateral or asymmetric PEX syndrome? An electron microscopy study]. AB - BACKGROUND: The pseudoexfoliation (PEX) syndrome which is characterized by the accumulation of an abnormal extracellular material in intra- and extraocular tissues, clinically strictly manifests both unilateral and bilateral. However, the generalized nature of this matrix process does not support unilateral ocular manifestation. The aim of this study, therefore, was a detailed histopathological analysis of the apparently not involved fellow eyes in so-called "unilateral" PEX syndrome. MATERIAL AND METHODS: For transmission electron microscopy, 5 pairs of donor eyes with slitlamp-microscopic, macroscopic and light microscopic evidence of unilateral PEX syndrome and 6 normal control eyes were studied. For immunohistochemistry, light and electron microscopic antibodies against LTBP-1 and HNK-1, two well-known markers for PEX deposits, were used. RESULTS: All apparently not involved contralateral eyes showed ultrastructural changes in the iris, in the ciliary body and in the trabecular meshwork. These changes include deposits of typical PEX fibrils on iris and ciliary epithelia as well as in the iris dilator muscle, microfibrillar precursors in the periphery of iris vessels, degenerative changes of the iris pigment epithelium and of the dilator muscle and an increased accumulation of extracellular matrix components around iris vessels, in the dilator muscle and in the juxta-canalicular connective tissue of the trabecular meshwork. In all contralateral and PEX eyes, but not in the control eyes, LTBP-1 and HNK-1 positive deposits could be identified in the periphery of iris vessels and in the dilator by light- and electron microscopic immunolabeling. CONCLUSIONS: The observed alterations in contralateral eyes in so called "unilateral" PEX syndrome support the concept that the PEX syndrome is a generalized, basically bilateral disease which may present in a clinically asymmetric manifestation. This should be considered in the clinical management of these patients. PMID- 11022665 TI - [Painless orbital swelling after sojourn in tropics. Cysticercosis and other parasitic eye diseases]. AB - BACKGROUND: The differential diagnosis of painless orbital swelling is complex and based on radiological and physical examination as well as laboratory tests. Due to increasing tourism to exotic countries a thorough history is important to observe diseases which are rare in industrialised countries but frequent in developing countries. PATIENT: A 30-year-old man complained about a painless orbital swelling in the absence of general symptoms. Orbital examination revealed a normal globe. MRI scan and B-scan ultrasounds showed an orbital cyst with a diameter of 1 cm. The patient's history disclosed lengthy travel to India as well as South East Asia. RESULTS: Histopathological examinations following excision of a whitish intraorbital mass showed a cream-white, thin walled cyst with a single central invaginated scolex with suckers and hooklets. This finding is characteristic for cysticercosis. CONCLUSIONS: Cysticercus cellulose is the larval stage of the pork tapeworm Taenia solium. It is transmitted by ingestion of contaminated food or water. Ocular affection is reported concerning the vitreous or subretinal localisation, less often in the anterior segment, subconjunctiva or orbita. Especially after trips to tropical regions, parasitic diseases should be kept in mind when dealing with unusual eye symptoms. Besides cysticercosis there are e.g. echinococcosis, onchocercosis (river blindness) and infections with toxocara or Loa loa. PMID- 11022666 TI - [Late diagnosis of Morquio syndrome. Clinical histopathological findings in a rare mucopolysaccharidosis]. AB - BACKGROUND: The Morquio syndrome is a rare autosomal-recessive mucopolysaccharidosis. The Morquio syndrome is characterized by a reduced activity of N-acetylgalactosamine-6-sulfate-sulfatase (type A), or beta galactosidase (type B). This deficiency leads to a lysosomal storage disease with accumulation of keratan sulfate und chondroitin-6-sulfate in connective tissue, skeletal system und teeth. Consequently, abnormalities of the skeletal system, aortic valvular disease and dental abnormalities occur. Ophthalmologically, diffuse corneal opacification and alterations of the trabecular meshwork- occasionally leading to glaucoma--can be found. CASE REPORT: A 44-year-old woman asked for perforating corneal transplantation because of corneal clouding on both eyes. Besides, she suffered from dwarfism of unclear reason. The diffuse corneal clouding and the dwarfism suggested a systemic-metabolic disease. Thus, further radiologic and medical investigation was started. RESULTS: Radiologically, a kyphoscoliosis, a pectus carinatum, a luxation of both hips, and a gonarthrosis were recognized. Fibroblast culture of a skin biopsy showed reduced activity of N acetyl-galactosamine-6-sulfate-sulfatase. This was the proof of Morquio syndrome type A. The explanted corneal button showed granules of acid mucopolysaccharides. Those were in the epithelial and endothelial cells and in the corneal stroma. CONCLUSIONS: To our knowledge, a Morquio syndrome has never been diagnosed with an adult and only after the ophthalmologist gave a hint. This can only be explained by the comparative mild expression of the disease in this patient. Diagnosis of Morquio syndrome is important because the frequent odontoid hypoplasia can lead to a deadly atlanto-axial instability, if not treated. PMID- 11022667 TI - [Immunophenotype classification of macular corneal dystrophy: first case report of immunophenotype I A outside of Saudi Arabia. A clinical histopathological correlation with immunohistochemistry and electron microscopy]. AB - BACKGROUND: Macular corneal dystrophy can be classified into three different immunophenotypes according to presence and distribution of sulfated keratan sulfate (SKS) in the cornea and serum levels of SKS. Immunophenotype I A has yet only been reported in patients with macular corneal dystrophy from Saudi Arabia. MATERIAL AND METHODS: Case report of a patient with macular corneal dystrophy type I A from Germany. Indirect immunohistochemistry for sulfated keratan sulfate was performed on the corneal buttons obtained after perforating keratoplasty (monoclonal antibodies 3D12/H7 and 5-D-4). The serum concentration of SKS was determined using the monoclonal antibody 5-D-4 in a serum antigen-inhibition assay. Immunogold labeling with the 5-D-4 antibody was performed to allow ultrastructural localization of SKS in the cornea. RESULTS: The patient demonstrated the typical clinical picture of macular corneal dystrophy. Serum levels of SKS were not detectable. Positive immunohistochemistry for SKS was found only within single keratocytes of both corneae without staining of the stroma (3D12/H7 and 5-D-4). Immunogold labeling localized SKS within stromal keratocytes and proved this patient to suffer from macular corneal dystrophy immunophenotype I A. CONCLUSION: Macular corneal dystrophy immunophenotype I A can be found outside Saudi Arabia in a German patient. All three immunophenotypes of macular corneal dystrophy are present in corneal buttons from German patients. PMID- 11022668 TI - [Acute bilateral angle-closure glaucoma secondary to ciliary body epithelial cysts of the pars plicata--correlation with the ultrasound biomicroscope]. PMID- 11022670 TI - [Eyelid tumor as a recurrence of a gastric MALT-lymphoma]. AB - BACKGROUND: MALT Lymphoma of the conjunctiva, eyelid and orbit represents a primary manifestation of the disease in most of the cases. CASE REPORT: A 58-year old male presented with a tumor in the left inferior eyelid. A full-thickness incisional biopsy was performed. Histopathologically, a lymphoid infiltrate involved the eyelid in its full width, with reactive follicles surrounded by small cells with irregular nucleus. Follicular colonization and lymphoepithelial lesions were detected. The tumor was CD20 positive. The molecular genetic analysis disclosed a monoclonal B-cell proliferation. The patient had a history of gastric MALT lymphoma treated 9 years ago and he had been otherwise in good health. The histopathologic features of the gastric tumor were identical to those of the eyelid lesion. Since the eyelid tumor represented a recurrence of the disease, treatment consisted of systemic chemotherapy. Complete tumor eradication was obtained. CONCLUSION: Ocular adnexal MALT-lymphomas may represent a recurrence of a distant primary MALT-lymphoma after a long disease-free period and, therefore, be an indication for systemic chemotherapy. PMID- 11022669 TI - [Brainstem anesthesia with respiratory arrest after retrobulbar block--a case report with a review of literature]. AB - BACKGROUND: This case shows the time course and typical clinical features of brain stem anaesthesia. It is presumed that it follows accidental injection of local anaesthetic into the subarachnoidal space via the optic nerve sheaths. HISTORY AND SIGNS: We report the case of a 70-year-old man, who became unconscious shortly after retrobulbar anaesthesia. The first symptoms developed after an interval of several minutes following the injection of the local anaesthetic. After the appearance of cranial nerves deficits the patient complained of difficulty breathing and became unconscious and apnoeic. THERAPY AND OUTCOME: It was decided to intubate the patient. Heart rate and systolic blood pressure dropped, therefore, a central line was placed and intravenous fluids were given. The patient was brought to the intensive care unit where he was extubated uneventfully the same day. CONCLUSIONS: The quick clinical recovery of the patient without any sequelae and the unremarkable results of internal and neurological examines support the diagnosis of brain stem anaesthesia. PMID- 11022672 TI - [Infection and regulation of leukocytes by autonomic nervous system]. AB - The distribution of leukocyte pattern (i.e., the granulocyte to lymphocyte ratio) is changed by microbial infections. We found that the autonomic nervous system is also intimately associated with the inflammatory response. Even in the absence of such infections, the distribution of leukocyte pattern is changed by stimulation of the autonomic nervous system itself. When these changes become significant, people are susceptible to diseases such as purulent inflammation or allergic inflammation. PMID- 11022673 TI - [Transplantation surgery and immune regulation]. AB - Although transplantation has revolutionized care for patients with end-stage organ failure due to the development of new immunosuppressive drugs, significant problems persist with treatments designed to prevent acute graft rejection. There is a real need for improved methods to maintain allograft function by preventing immune-mediated rejection. Accumulating evidence suggests that the inhibition of T cell costimulation may promote antigen-specific tolerance of transplants. In particular, blockade of the CD28 and CD40 costimulatory pathways has shown great promise in preventing transplant rejection in rodents and nonhuman primates. This paper discusses the human immune system, costimulatory pathways, and new therapeutic agents that regulate so-called costimulatory second signaling to maintain transplant function without general suppression of the immune system. PMID- 11022671 TI - [Cytokine production in surgical stress]. AB - Surgical injury influences the function of mononuclear cells, leading to various systemic responses. Proinflammatory cytokines such as tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1, -6, -8, and the antiinflammatory cytokine IL-10, which are mainly produced by mononuclear cells, are known to play an important role in the response to and pathogenesis of surgical stress. TNF alpha production by monocytes is extremely upregulated, but monocyte HLA-DR antigen expression is suppressed in patients with surgical stress. While production of Th1 cytokines such as IL-12 and interferon-gamma by mononuclear cells is suppressed, production of Th2 cytokines and IL-10 is upregulated during surgical stress. Immune suppression following surgical stress has been clarified recently in terms of Th1 and Th2 cytokine production balance mainly caused by mononuclear cells. It is thought to be very important to maintain immunological function after surgical stress by controlling cytokine production and balance. PMID- 11022674 TI - [Immunoguided surgery]. AB - Radioimmunoguided surgery has been attempted using intraoperative tumor detection with radiolabeled monoclonal antibody (mAb). Directly radiolabeled mAb showed poor tumor imaging due to high blood-pool radioactivity. In 1991, three-step immunoscintigraphy of the avidin-biotin system was reported. This three-step method improved tumor imaging because it reduced background radioactivity. Although there were some problems such as a human anti-mouse response and anti avidin response, the three-step clinical protocol was performed in various cancer patients, and many good results (the tumor detection rate was about 85%) were reported. On the other hand, recent major trials on tumor localization and sentinel lymph node detection have been performed using FDG-PET. However, radioimmunoscintigraphy has the major advantage of simultaneous tumor diagnosis and treatment. In the near future, radioimmunoscintigraphy may become a useful diagnostic method with anticancer effects when its adverse effects are ameliorated by the application of gene technology. PMID- 11022675 TI - [Antibody treatment for cancer]. AB - The previous pessimistic view of monoclonal antibodies as cancer therapeutics was largely influenced by the negative results of multiple clinical trials initiated shortly after the introduction of hybridoma technology. Although murine antibodies are of value in cancer therapy, their effectiveness is limited because murine monoclonal antibodies have a short survival time in humans and induce an immune response that neutralizes their therapeutic effect. Furthermore, the responses induced by murine antibodies are limited because they only weakly recruit human effector elements. To circumvent these problems, genetically engineered antibodies were produced which were mouse-human chimeric monoclonal antibodies and humanized antibodies. These antibodies combine the murine variable or hypervariable regions with the human constant or constant and variable framework regions. Numerous published studies demonstrate that erbB-2 oncogene product overexpression is a frequent cause of breast, ovarian, lung, and gastric cancers. Recently, an anti-erbB-2 humanized monoclonal antibody has been generated and used in clinical trials with high clinical response rates. We generated the anti-erbB-2 mouse-human chimeric monoclonal antibody CH401, which was able to kill cancer cells. An analysis of the mechanism of its tumor growth inhibition revealed that the cytotoxicity was induced by apoptosis. This cytotoxic mechanism is not the same a that of other anti-erbB-2 antibodies previously reported. PMID- 11022676 TI - [Human tumor-rejection antigens and peptides from genes to clinical research]. AB - One of the most significant advances in the field of modern tumor immunology is the identification of genes encoding tumor-rejection antigens that are recognized by human leukocyte antigen (HLA) class I-restricted and tumor-specific cytotoxic T lymphocytes (CTLs). Several peptides encoded by these genes are now under clinical trial as cancer vaccines, and major tumor regression has been observed in some melanoma patients. These results indicate that identification of the peptides capable of inducing CTLs may provide a new modality of cancer therapy. We investigated tumor-rejection antigens from epithelial cancers, and reported 7 genes encoding tumor-rejection antigens and peptides available for specific immunotherapy of HLA-A26 or -A24 patients with epithelial cancers. Furthermore, we identified more than 10 genes encoding tumor-rejection antigens and peptides available for specific immunotherapy of HLA-A2 patients with epithelial cancers. Therefore these new antigens and peptides could be applicable to the treatment of numerous epithelial cancer patients worldwide. Phase I clinical trials of cancer vaccine with these peptides for epithelial cancer patients are in progress at our university. Basic and clinical research will provide new insights for a better understanding of the molecular basis of T cell-mediated recognition of cancer cells and be important for the development of cancer vaccines. PMID- 11022677 TI - [Review of cancer gene therapy]. AB - Since the first introduction of gene-marking technology to the clinical field in 1989 by Rosenberg et al, more than 4,000 patients have participated gene therapy clinical trials worldwide. Most of those patients had malignancies. Nearly 90% of clinical trials, however, are still in phase I-II stage, and only 3 protocols are in the phase III stage in early 2000. As current clinical gene therapy protocols are intended essentially to examine the safety and feasibility of the new strategy, more careful and steady steps may be required before these clinical trials really produce clinical benefits. Focused on cancer gene therapy, direct and indirect approaches are undertaken. In the direct approach, HSV-TK, HLA-B7, or p53 tumor suppressor gene therapies are the three major approaches historically. In for the indirect approach, cytokine or adhesion molecule gene transferred tumor cells or immunocompetent cells are considered to be promising to enhance patients' antitumor immunity. In particular, we have concentrated on developing immuno gene therapy using GM-CSF-transduced autologous tumor cells. We have already recruited three patients with stage IV renal cell cancer. In all patients, peripheral blood T cells were mobilized after vaccination with GM-CSF transduced tumor cells, and two of the three patients showed the persistence of cytotoxic T cells against autologous tumor cells. Clinically, one patient has been followed up with stable disease for more than one year since the start of vaccination. Further clinical studies are required to obtain conclusive results. PMID- 11022678 TI - [Epidemiological trends of prostate cancer in Japan]. PMID- 11022679 TI - [International comparisons of prostate cancer incidence and mortality]. PMID- 11022680 TI - [Natural history of prostatic cancer]. PMID- 11022681 TI - [Prostate cancer and androgen]. PMID- 11022682 TI - [Mutations of oncogenes and tumor suppressor genes]. PMID- 11022683 TI - [The mechanism of hormone-independent recurrence of prostate cancer]. PMID- 11022684 TI - [Risk factors for prostate cancer and perspective on its primary prevention]. PMID- 11022685 TI - [Histopathological classification of prostate cancer]. PMID- 11022686 TI - [Recent advances in prostate pathology]. PMID- 11022687 TI - [Clinical staging classification]. PMID- 11022688 TI - [The advancement of diagnostic modalities for prostate cancer]. PMID- 11022689 TI - [An outline of the process of diagnosis]. PMID- 11022690 TI - [Symptoms, physical examination and digital rectal examination]. PMID- 11022691 TI - [Diagnosis of prostate cancer in general practice]. PMID- 11022692 TI - [Prostate specific antigen: a role of PSA in the diagnosis of prostate cancer]. PMID- 11022693 TI - [Free/total PSA ratio]. PMID- 11022694 TI - [PSA density, PSA velocity and PSA doubling time]. PMID- 11022695 TI - [Prostate specific antigen adjusted for the transition zone volume]. PMID- 11022696 TI - [Present status of serum total PSA assay and factors responsible for the inter kit deviations]. PMID- 11022697 TI - [PSA-ACT]. PMID- 11022698 TI - [Development of diagnostic imagings for prostatic cancer and their detection rates]. PMID- 11022699 TI - [Clinical significance of transrectal ultrasonography in screening and staging for prostate cancer]. PMID- 11022700 TI - [CT and MRI]. PMID- 11022701 TI - [Bone scintigraphy]. PMID- 11022702 TI - [Current advances of needle biopsy techniques of the prostate]. PMID- 11022703 TI - [Systematic random core biopsy of the prostate]. PMID- 11022704 TI - [Detection of PSA mRNA and PSMA mRNA by means of RT-PCR]. PMID- 11022705 TI - [Current value of prostate specific membrane antigen]. PMID- 11022706 TI - [Insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) as serum markers of prostate cancer]. PMID- 11022707 TI - [Carboxyterminal propeptide of type I procollagen (PICP), cross-linked carboxyterminal telopeptide of type I collagen (ICTP)]. PMID- 11022708 TI - [Management of prostate cancer-by-stage]. PMID- 11022710 TI - [Surgical castration]. PMID- 11022709 TI - [Evolution of endocrine therapy for prostate cancer]. PMID- 11022711 TI - [Goserelin acetate depot, LH-RH agonist; its properties and therapeutic effects in prostate cancer]. PMID- 11022712 TI - [Leuprorelin acetate depot]. PMID- 11022713 TI - [Total androgen blockade therapy]. PMID- 11022714 TI - [Adverse reaction of LH-RH agonist and its countermeasure]. PMID- 11022716 TI - [Basic and clinical characteristics of flutamide]. PMID- 11022717 TI - [Bicalutamide]. PMID- 11022715 TI - [Usefulness and problems of antiandrogen]. PMID- 11022718 TI - [Revaluation of estrogen therapy on prostate cancer]. PMID- 11022719 TI - [Chemotherapy for prostate cancer]. PMID- 11022720 TI - [History of surgical treatment for prostate cancer]. PMID- 11022721 TI - [Radical prostatectomy for locally advanced adenocarcinoma of the prostate]. PMID- 11022722 TI - [Nerve-sparing radical prostatectomy]. PMID- 11022723 TI - [Troubles and troubleshootings in radical prostatectomy]. PMID- 11022724 TI - [Progress in radiation therapy for prostate cancer]. PMID- 11022726 TI - [Brachytherapy]. PMID- 11022725 TI - [Liniac irradiation]. PMID- 11022727 TI - [Heavy ion radiotherapy]. PMID- 11022728 TI - [Proton beam radiotherapy]. PMID- 11022729 TI - [Palliative radiotherapy (bone metastasis and lymph node metastasis)]. PMID- 11022730 TI - [Recto-vesical morbidity in radiation therapy for prostate cancer]. PMID- 11022731 TI - [Intermittent androgen suppression for prostate cancer]. PMID- 11022732 TI - [Step up therapy]. PMID- 11022733 TI - [Peptide-pulsed dendritic cell therapy]. PMID- 11022734 TI - [Gene therapy]. PMID- 11022735 TI - [High intensity focused ultrasound for localized prostate cancer]. PMID- 11022736 TI - [Cryosurgery]. PMID- 11022737 TI - [Long-acting depot of LH-RH agonist]. PMID- 11022738 TI - [Inhibitors of the key enzymes of androgen synthesis: potential agents as targets for prostate cancer]. PMID- 11022739 TI - [New anti-androgen and luteinizing hormone releasing hormone antagonist for the treatment of advanced prostate cancer]. PMID- 11022740 TI - [Neoadjuvant androgen ablation therapy (NAT)]. PMID- 11022741 TI - [Chemo-endocrine therapy]. PMID- 11022743 TI - [High-dose intravenous estrogen therapy]. PMID- 11022742 TI - [Real-time tumor-tracking radiotherapy combined with neoadjuvant hormonal therapy for prostate cancer]. PMID- 11022744 TI - [Estramustine phosphate and other chemotherapeutic agents in hormone refractory prostate cancer]. PMID- 11022745 TI - [Treatment of hormone-refractory prostate cancer with etoposide (Vp16)]. PMID- 11022746 TI - [Dexamethasone for hormone refractory prostate cancer]. PMID- 11022747 TI - [Biochemical modulation and kampo therapy]. PMID- 11022748 TI - [New horizon for PET imaging]. PMID- 11022749 TI - [Serum and urinary PSA in the diagnosis of PSA failure after radical prostatectomy]. PMID- 11022750 TI - [Mass screening for prostate cancer]. PMID- 11022751 TI - [Health screening for prostate cancer]. PMID- 11022752 TI - [Cost-effectiveness of prostatic cancer screening]. PMID- 11022753 TI - [Hormone therapy and QOL in prostate cancer patients]. PMID- 11022754 TI - [Radical prostatectomy, procedure and quality of life]. PMID- 11022755 TI - [Palliative therapy]. PMID- 11022756 TI - [Informed consent in prostate cancer screening]. PMID- 11022757 TI - [Informed consent of prostate cancer management]. PMID- 11022758 TI - [Familial prostate cancer and prostate cancer susceptibility locus on chromosome 1 (hereditary prostate cancer-1, HPC-1)]. PMID- 11022759 TI - [Discrepancies between Japanese general rules of prostatic cancer and Gleason grading system]. PMID- 11022760 TI - [Is cure of T1c prostate cancer necessary in those for whom it is possible?]. PMID- 11022761 TI - [In vivo ultrathin endoscopy of seminal vesicles for prostate cancer]. PMID- 11022762 TI - [Evaluation of prostate cancer with FDG PET study after therapy]. PMID- 11022763 TI - [Telomerase activity in prostate cancer]. PMID- 11022764 TI - [Treatment of prostate cancer in elderly patients]. PMID- 11022765 TI - [Relation of antiandrogen withdrawal syndrome to androgen receptor]. PMID- 11022766 TI - [Side effects and quality of life in prostate cancer following treatment]. PMID- 11022767 TI - Detection of major putative periodontopathogens in Korean advanced adult periodontitis patients using a nucleic acid-based approach. AB - BACKGROUND: Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systematic analysis of subgingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to describe the prevalence of major putative periodontopathogens in Korean patients by culture-independent methods. METHODS: A total of 244 subgingival plaque samples (5 sites in each participant) were taken from 29 advanced adult periodontitis (AP) patients and 20 periodontally healthy subjects. AP samples were obtained from the 4 deepest periodontal pockets (> or =6 mm probing depth [PD]) and 1 healthy site (< or =3 mm PD) in each patient. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed with eubacterial primers. Aliquots of PCR products were then applied on nylon membranes and hybridized with specific oligonucleotide probes labeled with digoxigenin. RESULTS: All diseased sites harbored Fusobacterium sp., while Porphyromonas gingivalis, Treponema sp., and Bacteroides forsythus were detected in more than 96% of 116 diseased sites. Peptostreptococcus micros, Actinobacillus actinomycetemcomitans, and Prevotella intermedia were present in 82%, 74%, and 71% of diseased sites, respectively. In sites of periodontally healthy subjects, Fusobacterium sp. was present in the highest proportion (58%). Treponema sp., P. gingivalis, and B. forsythus were detected in 22%, 18%, and 18% of healthy sites, respectively. P. micros, P. intermedia, and A. actinomycetemcomitans were found in 8%, 2%, and 1% of healthy sites, respectively. The prevalence of the periodontopathogens, with the exceptions of Fusobacterium sp. and B. forsythus, was significantly higher in the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects (P <0.05). CONCLUSIONS: Using highly sensitive methods relying on 16S ribosomal RNA-based oligonucleotide probes, we confirmed the strong association of 7 putative periodontopathogens with AP patients in a Korean population. With the exceptions of Fusobacterium sp. and B. forsythus, all the periodontopathogens were significantly more associated with the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects. PMID- 11022768 TI - HLA-B27 transgenic rats are susceptible to accelerated alveolar bone loss. AB - BACKGROUND: HLA-B27 transgenic rats exhibit generalized, severe inflammatory reactions and spontaneously develop arthritis and chronic gastrointestinal inflammation, as well as inflammatory lesions in other tissues. Our hypothesis was that HLA-B27 rats would also be susceptible to inflammatory periodontal disease, and therefore alveolar bone loss. The purpose of this investigation was to compare the naturally occurring alveolar bone loss in HLA-B27 and wild type rats. METHODS: Age- and sex-matched HLA-B27 transgenic (TG) and wild type Fischer 344 (WT) female retired breeders, and their age-matched male WT breeding mates, were examined for alveolar bone loss (ABL). Thirty-eight animals were used: twelve, 20, and 6 animals were 6, 8, and 12 months old, respectively. ABL was measured as the exposed root surface area (mm2) in the defleshed maxilla and mandible. RESULTS: The coefficient of variation for replicate ABL measurements was 4.4%. For the 6- and 8-month age groups, ABL was significantly greater in TG rats compared to WT rats. The observed difference in ABL between TG and WT animals did not reach statistical significance for the 12-month age group. Within each of the two animal groups (TG and WT), ABL was significantly different between age groups. The ABL rate of TG female rats was 42% to 250% greater than that of WT female rats, depending on the age range examined. CONCLUSIONS: HLA-B27 rats are susceptible to accelerated alveolar bone loss and could serve as an animal model of alveolar bone loss pathogenesis. PMID- 11022769 TI - Morphology and elemental composition of subgingival calculus in two ethnic groups. AB - BACKGROUND: The aim of the present study was to compare the morphology and elemental composition of subgingival calculus between Indo-Pakistani and Caucasian patient groups. METHODS: Extracted teeth from 14 Indo-Pakistani and 19 Caucasian subjects were collected. Of these, 12 Indo-Pakistani and 10 Caucasian teeth had sufficient subgingival calculus for analysis. Subgingival calculus present on the 22 teeth was classified into six morphological types: 1) crusty/spiny/nodular; 2) ledge/ ring; 3) thin, smooth veneers; 4) finger/fern like; 5) individual islands/spots; or 6) supramarginal on submarginal. Subgingival calculus was zoned: coronal, mid, and apical. A sample obtained from each zone was subdivided to allow 3 separate analyses: transmission electron microscopic (TEM) x-ray microanalysis for elemental composition, fluoride analysis, and carbonate analysis. RESULTS: Crusty/spiny/nodular, ledge/ring, and thin, smooth veneers were more commonly found in the Indo-Pakistani group; individual islands were more prevalent in the Caucasian subjects. Supramarginal on submarginal calculus was found only in the Indo-Pakistani group. No finger/fern-like deposits were found. No differences within or between the two ethnic groups were found with regard to calcium:phosphate ratios, fluoride, or carbonate content. However, the Indo-Pakistani group showed significantly lower levels of sodium in apical samples than in coronal samples (ANOVA, F1,16 = 5.98, P= 0.03), and significantly lower levels of sodium (ANOVA, F1,12 = 4.75, P= 0.05) and magnesium (ANOVA, F1,12 = 5.16, P= 0.04) in apical samples than in those from Caucasians. After adjusting for smoking, the magnesium results remained significant (ANOVA, F2,11 = 4.64, P= 0.05). CONCLUSIONS: Subgingival calculus demonstrated differences in morphology between these two ethnic groups and differences in elemental composition, which may influence its overall solubility and contribute to its greater accretion in the Indo-Pakistani subjects. PMID- 11022770 TI - Crestal bone changes around titanium implants. A histometric evaluation of unloaded non-submerged and submerged implants in the canine mandible. AB - BACKGROUND: Today, implants are placed using both non-submerged and submerged approaches, and in 1- and 2-piece configurations. Previous work has demonstrated that peri-implant crestal bone reactions differ radiographically under such conditions and are dependent on a rough/smooth implant border in 1-piece implants and on the location of the interface (microgap) between the implant and abutment/restoration in 2-piece configurations. The purpose of this investigation was to examine histometrically crestal bone changes around unloaded non-submerged and submerged 1- and 2-piece titanium implants in a side-by-side comparison. METHODS: A total of 59 titanium implants were randomly placed in edentulous mandibular areas of 5 foxhounds, forming 6 different implant subgroups (types A F). In general, all implants had a relatively smooth, machined coronal portion as well as a rough, sandblasted and acid-etched (SLA) apical portion. Implant types A-C were placed in a non-submerged approach, while types D-F were inserted in a submerged fashion. Type A and B implants were 1-piece implants with the rough/smooth border (r/s) at the alveolar crest (type A) or 1.0 mm below (type B). Type C implants had an abutment placed at the time of surgery with the interface located at the bone crest level. In the submerged group, types D-F, the interface was located either at the bone crest level (type D), 1 mm above (type E), or 1 mm below (type F). Three months after implant placement, abutment connection was performed in the submerged implant groups. At 6 months, all animals were sacrificed. Non-decalcified histology was analyzed by evaluating peri-implant crestal bone levels. RESULTS: For types A and B, mean crestal bone levels were located adjacent (within 0.20 mm) to the rough/smooth border (r/s). For type C implants, the mean distance (+/- standard deviation) between the interface and the crestal bone level was 1.68 mm (+/- 0.19 mm) with an r/s border to first bone-to-implant contact (fBIC) of 0.39 mm (+/- 0.23 mm); for type D, 1.57 mm (+/- 0.22 mm) with an r/s border to fBIC of 0.28 mm (+/- 0.21 mm); for type E, 2.64 mm (+/- 0.24 mm) with an r/s border to fBIC of 0.06 mm (+/- 0.27 mm); and for type F, 1.25 mm (+/- 0.40 mm) with an r/s border to fBIC of 0.89 mm (+/- 0.41 mm). CONCLUSIONS: The location of a rough/smooth border on the surface of non-submerged 1-piece implants placed at the bone crest level or 1 mm below, respectively, determines the level of the fBIC. In all 2-piece implants, however, the location of the interface (microgap), when located at or below the alveolar crest, determines the amount of crestal bone resorption. If the same interface is located 1 mm coronal to the alveolar crest, the fBIC is located at the r/s border. These findings, as evaluated by non-decalcified histology under unloaded conditions, demonstrate that crestal bone changes occur during the early phase of healing after implant placement. Furthermore, these changes are dependent on the surface characteristics of the implant and the presence/absence as well as the location of an interface (microgap). Crestal bone changes were not dependent on the surgical technique (submerged or non-submerged). PMID- 11022771 TI - The Fcgamma receptor genotype as a risk factor for generalized early-onset periodontitis in Japanese patients. AB - BACKGROUND: Genetic polymorphisms of immunoglobulin G (IgG) Fc receptors (FcgammaR) were recently shown to be associated with recurrence rates of adult periodontitis (AP). The purpose of this study was to evaluate whether FcgammaR polymorphisms are also associated with generalized early-onset periodontitis (G EOP) in Japanese patients. METHODS: Thirty-eight Japanese patients with G-EOP and 83 Japanese patients with AP were identified according to established clinical criteria, including measurements of probing depth, clinical attachment level, and alveolar bone level. FcgammaR genotypes for 3 bi-allelic polymorphisms were determined in these G-EOP and AP patients and 104 race-matched healthy controls by means of allele-specific polymerase chain reactions. RESULTS: There was a significant difference in the distribution of FcgammaRIIIb genotypes between G EOP patients and healthy controls (P = 0.02). Additionally, a significant over representation of FcgammaRIIIb-NA2 allele was observed in G-EOP patients as compared to AP patients and controls (P= 0.02, P= 0.009, respectively). Moreover, we found a strong association between G-EOP and the composite genotype comprising FcgammaRIIIb-NA2 and FcgammaRIIIa-158F (G-EOP versus controls: odds ratio 2.4, 95% CI 1.0-6.0, chi2 = 4.13, P= 0.04). CONCLUSIONS: This study indicates that the FcgammaRIIIb-NA2 allele and possibly FcgammaRIIIa-158F could be associated with susceptibility to G-EOP in Japanese patients. PMID- 11022772 TI - Evaluation of a new bioabsorbable barrier for recession therapy: a feasibility study. AB - BACKGROUND: The objective of this study was to evaluate the feasibility of a new polylactic acid bioabsorbable barrier in the treatment of gingival recession. METHODS: Twenty patients with buccal recession defects (Miller class I, II, and III; mean recession: 4.0 +/- 1.2 mm; range: 2.0 to 6.8 mm) participated. After thorough scaling and root surface conditioning with 10% tetracycline-HCl, a trapezoidal mucoperiosteal flap was prepared. A customized barrier was applied to cover the defect. Barriers adhered directly to tooth and bone and no sutures were used. The barrier was subsequently covered by a coronally positioned flap. Assessments of probing depths were performed by means of a controlled-force electronic probe, and recession was determined on stone models with a digital caliper at baseline and 12 months following therapy. RESULTS: Eight barriers showed limited exposure (1 to 2 mm) with minimal signs of gingival inflammation between 2 and 6 weeks following surgery. Comparing baseline measurements with outcomes at 12 months, significant root coverage and probing attachment gain were observed (P <0.0001, paired t test). Mean gingival recession was reduced to 0.4 +/- 0.5 mm, corresponding to a mean root coverage of 91.9%, and overall attachment gains amounted to 4.2 mm. A significant gain of keratinized tissue was found (2.9 +/- 0.7 mm), and mean probing depths were slightly reduced from 2.2 to 1.7 mm. CONCLUSIONS: The results of this study indicate favorable outcomes after using a new bioabsorbable barrier material for root coverage in recession-type defects. PMID- 11022773 TI - Treatment of gingival recession: comparative study between subepithelial connective tissue graft and guided tissue regeneration. AB - BACKGROUND: Various procedures have been proposed to treat gingival recession, but few studies compare these procedures to each other. The purpose of this study was to evaluate a clinical comparison of subepithelial connective tissue graft (SCTG) and guided tissue regeneration (GTR) with a collagen membrane in the treatment of gingival recessions in humans. METHODS: Twenty-four defects were treated in 12 patients who presented canine or pre-molar Miller Class I and/or II bilateral gingival recessions. Both treatments were performed in all patients, and clinical measurements were obtained at baseline and 18 months after surgery. These clinical measurements included gingival recession height (GR), root coverage (RC), probing depth (PD), keratinized tissue width (KT), and final esthetic result. RESULTS: Both SCTG and GTR with a bioabsorbable membrane and bone graft demonstrated significant clinical and esthetic improvement for gingival recession coverage. The SCTG group was statistically significantly better than GTR for height of GR (SCTG = 0.2 mm, GTR = 1.12 mm, P= 0.02) and KT (SCTG = 4.58 mm, GTR = 2.5 mm, P<0.0001). However, PD was statistically significantly better for GTR than SCTG treatment (GTR = 1.66 mm, SCTG = 1.00, P= 0.01). The 2 procedures were statistically similar in root coverage (SCTG = 95.6%, GTR = 84.2%, P= 0.073). The esthetic condition after both treatments was satisfactory (P= 0.024). CONCLUSIONS: It was concluded that the gingival recessions treated with the SCTG group were superior for GR, RC, and KT clinical parameters, while GTR demonstrated better PD reduction. The final esthetic results were similar using both techniques. PMID- 11022775 TI - Histologic response to intraligament injections using a computerized local anesthetic delivery system. A pilot study in mini-swine. AB - BACKGROUND: Intraligamentary (periodontal ligament) injection has been used to locally control pain with minimal anesthetic dose. The purpose of the present study was to determine the histologic effect of using a computer-controlled local anesthesia delivery system (CCLADS) for the administration of a periodontal ligament injection in the mini-swine model system. METHODS: Thirty-two sites in 3 mini-swine pigs with healthy periodontium were selected. Each site was notched on the mesial-lingual line angle of the tooth to allow a reference for needle placement and provide a histologic marker. An anesthetic cartridge was used with a 30-gauge needle attached to the handpiece of the CCLADS. This provided a controlled flow rate for 2 minutes. The control sites were treated with the same needle in place for 20 seconds. The animals were sacrificed and provided observation periods of 1 hour, 1, 4, 7, 21, and 49 days. Specimens were stained with hematoxylin and eosin and trichrome. RESULTS: Twenty-four hours after injection, limited localized inflammation was present. By 7 days, the ligament appeared within normal limits. Apical migration of the junctional epithelium extended to the apical limit of the notch and was similar in test and control specimens. CONCLUSIONS: Within the limits of this study, the histologic results showed that tissue responses following intraligament injections using a CCLADS demonstrated limited inflammatory responses within the first 24 hours, which abated by 7 days postinjection. Similar migration of the junctional epithelium was seen in test and control specimens and was probably related to tooth injury rather than anesthetic injections. Further investigations are warranted using CCLADS to determine whether clinical benefits will translate in the human model system. PMID- 11022774 TI - Gingival fluid ciprofloxacin levels at healthy and inflamed human periodontal sites. AB - BACKGROUND: Polymorphonuclear leukocytes (PMNs) take up and accumulate ciprofloxacin. This may allow them to enhance the delivery of this agent to the inflamed periodontium. METHODS: Cross-sectional and longitudinal approaches were used to test the hypothesis. In the cross-sectional study, 7 periodontally healthy adults and 8 adults with untreated periodontitis were administered three doses of ciprofloxacin (500 mg bid). Gingival fluid (GF) and serum samples were obtained after 28 hours and analyzed by high-performance liquid chromatography (HPLC). In the longitudinal study, 8 adult periodontitis subjects were administered 500 mg ciprofloxacin bid for 8 days. After 28 hours, GF from 4 sites with 5 to 8 mm probing depths was sampled in each subject, serum samples were obtained, and 2 of the 4 sites were root planed. GF and serum were sampled again 7 days later (196 hours after the initial dose). RESULTS: The mean ciprofloxacin levels in the GF and serum of periodontally healthy subjects were 2.52 +/- 0.22 microg/ml and 0.47 +/- 0.05 microg/ml, respectively. In subjects with periodontitis, these levels were 2.69 +/- 0.44 microg/ml and 0.61 +/- 0.13 microg/ml, respectively. GF ciprofloxacin levels were significantly higher than corresponding serum levels in healthy and diseased subjects (P<0.01), but there were no significant differences in GF or serum levels between the 2 subject groups. Since GF flow was significantly higher at diseased sites, however, more ciprofloxacin was distributed to these sites than to healthy sites. In the longitudinal study, GF flow at 196 hours was 16% lower at root planed sites than at untreated control sites (P = 0.412). The minor decrease in this index of inflammation was accompanied by a small (9%), but statistically significant (P= 0.007), decrease in GF ciprofloxacin levels. CONCLUSIONS: GF ciprofloxacin levels decreased slightly at inflamed periodontal sites after root planing, but were significantly higher than serum levels even at healthy periodontal sites. Inflammation may enhance the distribution of ciprofloxacin to diseased sites, but it is not a major determinant of GF ciprofloxacin levels. PMID- 11022776 TI - The influence of nicotine on the bone loss rate in ligature-induced periodontitis. A histometric study in rats. AB - BACKGROUND: The present study investigated the possible influence of nicotine on the bone loss rate in the furcation region due to ligature-induced periodontitis in rats. METHODS: Twenty adult male Wistar rats were included. After anesthesia, the tooth was randomly assigned to receive the cotton ligature in the sulcular area, while the contralateral tooth was left unligated. The animals were randomly assigned to one of the following treatments, including daily intraperitoneal injections: group A, 2 microl/g body weight of saline solution; group B, 2 microl/g body weight of a nicotine solution with 0.13 microl of nicotine/ml of saline solution; group C, 2 microl/g body weight of a nicotine solution with 0.19 microl of nicotine/ml of saline solution; and group D, 2 microl/g body weight of a nicotine solution with 0.26 microl of nicotine/ml of saline solution. Thirty days later, the animals were sacrificed and the specimens routinely processed for serial decalcified sections. RESULTS: Intergroup analysis revealed greater bone loss in the ligated teeth of group B (1.01 +/- 0.61 mm2), group C (1.14 +/- 0.72 mm2), and group D (1.36 +/- 0.60 mm2) when compared with group A (0.64 +/- 0.62 mm2) (P <0.01). However, no statistically significant differences in bone loss were found among groups B, C, and D. In addition, no bone loss was observed for unligated teeth (P>0.01). CONCLUSIONS: Within the limits of the present study, nicotine enhanced the effects of the local components of periodontal disease in a non-dose-dependent way; nevertheless, the administration of nicotine did not produce periodontal bone loss by itself. PMID- 11022777 TI - Long-term treatment with subantimicrobial dose doxycycline exerts no antibacterial effect on the subgingival microflora associated with adult periodontitis. AB - BACKGROUND: The purpose of this study was to determine whether treatment with subantimicrobial dose doxycycline (SDD), 20 mg bid, exerted an antimicrobial effect on the microflora associated with adult periodontitis. METHODS: Following the approval of the protocol and informed consent forms by the respective IRBs at the University of Florida and West Virginia University, 76 subjects with adult periodontitis were entered and randomly assigned to receive SDD or placebo. A split-mouth design was utilized, with each subject receiving subgingival scaling and root planing (SRP) in two quadrants immediately following baseline data collection, while the remaining two quadrants were left unscaled (non-SRP). Microbial samples were collected prior to treatment, after 3, 6, and 9 months of treatment, and after 3 months of no treatment. The samples were examined by microscopy and by enumeration on selective and non-selective media. RESULTS: All treatments resulted in statistically significant decreases in the proportions of spirochetes and motile rods (P <0.05) and in an increase in the proportion of coccoid forms (P <0.0001) relative to baseline. No between-treatment differences were detected between the SDD and placebo treatments in either the SRP or non-SRP design, with the exception of the small and large spirochetal groups. The spirochetal proportions present in the SDD group were significantly lower (P<0.05) than the paired placebo group during the 9-month treatment and was preceded by a significant decrease (P<0.01) in the proportion of microbiologic sample sites that bled on probing. No between- treatment differences were detected in any of the other microbial parameters. CONCLUSION: The microbial differences observed were attributed to the anticollagenase and anti-inflammatory properties of SDD and not to an antimicrobial effect. PMID- 11022778 TI - Long-term use of subantimicrobial dose doxycycline does not lead to changes in antimicrobial susceptibility. AB - BACKGROUND: Adjunctive subantimicrobial dose doxycycline (SDD) with scaling and root planing leads to improved clinical parameters of adult periodontitis, but has raised questions about potential changes in antibiotic susceptibility of the host microflora. Our four studies assessed whether long-term SDD changes antibiotic susceptibility of the oral microflora in adults with periodontitis. METHODS: In studies 1 and 2, adult patients with periodontitis were randomized to receive SDD 10 mg qd, 20 mg qd, 20 mg bid, or placebo. In study 3, patients were randomized to receive SDD 20 mg bid or placebo. No medication was administered in study 4, a follow-up to study 3. Subgingival plaque samples were collected at baseline (all studies) and at 12, 15 to 18, and 24 months (study 1); 12, 18, and 27 months (study 2); 3, 6, and 9 months (study 3); and 3 months post-study 3 (study 4). Antimicrobial susceptibility of isolated bacteria was assessed by: 1) minimum inhibitory concentration (MIC) levels (studies 1 and 2); 2) cross resistance to non-tetracycline antibiotics (studies 2 and 3); and 3) the proportion of doxycycline-resistant isolates (studies 3 and 4). RESULTS: Organism MIC levels remained constant among all treatment groups at 18 and 24 months compared with baseline (study 1). Observed changes in susceptibility at 12 and 18 months for the 20 mg groups were attributed to the limited number of isolates tested (study 1). There were no statistically significant differences in the proportion of doxycycline-resistant isolates among treatment groups (studies 3 and 4), and no evidence of multi-antibiotic resistance (studies 3 and 4) or cross resistance (studies 2 and 3) at any timepoint. CONCLUSION: Long-term SDD does not contribute to changes in antibiotic susceptibility. PMID- 11022779 TI - Guided bone regeneration of large mandibular defects in a primate model. AB - BACKGROUND: An earlier publication from our laboratory described the use of guided bone regeneration to fill large bone voids in the mandible created through en bloc resection in primates. The present report is an embellishment of this paper and describes bone regeneration experiments in 18 adult male Macaca mulatta monkeys to determine how long membranes must be in position to promote guided bone regeneration. METHODS: Thirty-six lesions were created in the mandibles of 18 monkeys in a standardized mandibular defect of 8 x 19 mm. Reinforced ePTFE membranes were placed in the animals and held in place with mini screws and sutures for anywhere from 1 to 12 months. No material was added to the defect. In addition to clinical studies, digital subtraction radiology and fluorescent labeling with tetracycline and histomorphometry are described. RESULTS: The results indicate that no bone gain was observed in membranes exposed for 1 month or less, but bone gain (approximately well over 90% of defects) was observed at 12 months when membranes were left in situ for 2 to 12 months (P <0.0001). No significant difference in the amount of bone gained at 12 months was observed for membranes left in place for intervals ranging from 2 to 12 months. A significant correlation between the amount of bone gain observed at 3 and 12 months was observed (P <0.0001). CONCLUSIONS: Data therefore suggest that membranes left in situ for 1 month or less result in minimal bone gain compared with membranes left in place from 2 to 12 months. In addition, labeling and stained sections clearly showed that the bone produced after 2 months of membrane placement is mature. PMID- 11022780 TI - The relationship between bone mineral density and periodontitis in postmenopausal women. AB - BACKGROUND: Systemic bone loss has been proposed as a risk factor for periodontal disease; however, the relationship between these two diseases is still not clear. The objective of this study was to assess the relationship between systemic bone mineral density and periodontal disease, controlling for known confounders. METHODS: The study population included 70 postmenopausal Caucasian women aged 51 to 78 (mean +/- SD: 62.10 +/- 7.1 years). Skeletal bone mineral density (BMD) was assessed by dual energy x-ray absorptiometry (DXA) at the neck, trochanter, intertrochanter, Ward's triangle, and total regions of the femur, and from the anterior-posterior view of the lumbar spine. Periodontal disease severity was represented by clinical attachment loss (CAL) and interproximal alveolar bone loss (ABL). Other measures of periodontal status included probing depth (PD), supragingival plaque, gingival bleeding on probing, and calculus. DXA and oral examinations were performed by calibrated examiners. Partial correlation coefficients (r) were obtained from multiple linear regression analysis adjusting for age, age at menopause, estrogen supplementation, cigarette smoking, body mass index, and supragingival plaque. RESULTS: Mean ABL was significantly correlated with BMD of the trochanter (r =- 0.27), Ward's triangle (r = -0.26), and total regions of the femur (r = -0.25). Mean CAL appeared to be related to BMD consistently at all regions of the skeleton, although the association did not reach statistical significance. CONCLUSIONS: We can conclude that skeletal BMD is related to interproximal alveolar bone loss and, to a lesser extent, to clinical attachment loss, implicating postmenopausal osteopenia as a risk indicator for periodontal disease in postmenopausal Caucasian women. PMID- 11022781 TI - The relationship between peripheral levels of leukocytes and neutrophils and periodontal disease status in a patient with congenital neutropenia. AB - BACKGROUND: Congenital neutropenia is characterized by an almost total absence of neutrophils and increased susceptibility to infection. Oral manifestations include ulcerations of mucous membranes, acute gingival inflammation with focal necrosis, and rapid loss of attachment. Treatment with recombinant human granulocyte colony-stimulating factor (rhG-CSF) increases neutrophil numbers and ameliorates the periodontal status. METHODS: We report the treatment of a 22-year old male with congenital neutropenia (Kostman syndrome), referred to us due to periodontal disease, and the effect of treatment on peripheral neutropenia. Diagnosis of neutropenia was made at year 1; at age 15, the patient started to receive injections of rhG-CSF, reducing the occurrence of infection and improving neutrophil count, although levels remained below normal. The patient underwent extraction of a molar at age 8; scaling, root planing, and modified Widman flaps at age 9; and oral hygiene maintenance every 2 to 3 months from age 18 to 21. At age 23, he initiated treatment at our periodontal clinic. The patient's gingiva was severely inflamed, and the dentition was covered with plaque and calculus. Attachment loss was advanced, all teeth were mobile, and bone loss was approximately 75% in most sites. Neutrophil counts were below normal, but other hematologic parameters were normal. Scaling and root planing were performed and the patient received antibiotics and chlorhexidine rinses twice each day for 2 weeks. Extracoronal splinting was performed, fluoride varnish was used to desensitize cervical areas, and tooth FDI #46 was restored. Root planing and deplaquing were repeated, and the patient received subgingival chlorhexidine irrigation 13 times over one year. Assessments were made on presentation, after the initial treatment, and at 1 and 2 years post-treatment. RESULTS: Mean probing depth was reduced posttreatment with a further reduction during the maintenance period. This was correlated with an increase in attachment levels. Total white blood cells increased, due in part to an increase in neutrophils, reaching normal levels. CONCLUSIONS: This report demonstrates for the first time that periodontal therapy, resulting in decreased bacterial load, may result in restoration of normal levels of circulating neutrophils in individuals with congenital neutropenia under treatment with rhG-CSF. The results also suggest that periodontal pathogens may be associated with depressed neutrophil levels, even when patients receive treatment for neutropenia. PMID- 11022782 TI - Treatment of multiple recession-type defects in patients with esthetic demands. AB - BACKGROUND: When multiple recession defects affecting adjacent teeth in esthetic areas of the mouth are present, patient-related considerations suggest the selection of surgical techniques that allow all gingival defects to be simultaneously corrected with the soft tissue close to the defects themselves. The aim of the present study was to evaluate, with respect to root coverage, the effectiveness of a new surgical approach to the coronally advanced flap procedure for the treatment of multiple recession defects in patients with esthetic demands. METHODS: Twenty-two young systemically and periodontally healthy subjects with at least 2 recession-type defects affecting adjacent teeth in esthetic areas of the mouth were enrolled in the study. All recessions were Miller Class I or II. In each patient, all present recessions were treated at the same time with a modification of the coronally advanced flap technique. The clinical re-evaluation was made 1 year after the surgery. RESULTS: A total of 73 recessions (mean recession depth 2.8 mm) were treated; mean number of gingival recessions in each subject was 3.4. At the 1-year examination, on average, 97% of the root surface was covered with soft tissue and 64 defects (88%) showed complete root coverage. Complete root coverage in all recessions was achieved in 16 out of 22 patients (73%), and no statistically significant relationship was found between the root coverage results and the number of recession defects treated in each patient. A statistically significant increase of keratinized tissue (0.6 mm) was observed after 1 year; this increase was inversely correlated (P <0.001) with the amount of presurgical keratinized tissue. The multiple regression model showed that the final result, in terms of root coverage, was significantly affected by the initial recession depth and by the amount of presurgical keratinized tissue. Greater reductions in recession depth were observed in the cases with worse initial conditions and with lesser amount of keratinized tissue apical to the recession defect. CONCLUSIONS: The results of the present study demonstrated that the proposed surgical technique was very effective for the treatment of multiple gingival recessions affecting teeth in esthetic areas of the mouth and that these successful root coverage results could be achieved irrespective to both the number of recessions simultaneously treated during the surgical intervention and the presence, before surgery, of a minimal amount of keratinized tissue apical to the defects. PMID- 11022783 TI - A perspective on clinical significance. PMID- 11022784 TI - Re: Human periodontal ligament fibroblast behavior on chemically conditioned dentine: an in vitro study. Rompen EH, Goffinet GH, Nusgens B (1999;70:1144 1152). PMID- 11022785 TI - Evolution of viruses by acquisition of cellular RNA or DNA nucleotide sequences and genes: an introduction. AB - The origins of virus evolution may be traced to Archeabacteria since Inouye and Inouye (6) discovered a retroelement with a gene for reverse transcriptase in the bacterial genome and in the satellite, multiple copy single stranded DNA (msDNA) in the soil bacterium Myxococcus xanthus. It was possible (8) to define the evolution of retroelements in eukaryotic cells of plants, insects (gypsy retrovirus) and vertebrates. The replication of RNA viruses in eukaryotic cells allowed for the viral RNA genome to integrate a cellular ubiquitin mRNA, as reported for BVDV (24). Another example is the integration of 28S ribosomal RNA into the hemagglutinin gene of an influenza virus. This change in the hemagglutinin gene led to an increased pathogenicity of the influenza virus (25). In contrast to RNA viruses, DNA viruses had evolved by inserting cDNA molecules derived from mRNA transcripts of cellular genes or foreign viral RNA. It is of interest that the virus acquired cellular genes in the genomes of DNA viruses represent genes that code for proteins that inhibit cellular molecular processes related to HLA class I and II molecules. The other acquired genes are cellular genes that code for cytokines that are capable of inhibiting antigen presentation to T cells by antigen presenting cells (APC) by dendritic Langerhans cells. The acquisition of cellular genes by DNA viruses enhances their pathogenicity by inhibiting the hosts' defense systems. PMID- 11022786 TI - Immunomodulatory functions encoded by the E3 transcription unit of adenoviruses. AB - Persistent viruses have evolved multiple strategies to escape the host immune system. One important prerequisite for efficient viral reproduction in the face of an ongoing immune response is prevention of premature lysis of infected cells. A number of viruses achieve this goal by interfering with antigen presentation and recognition of infected cells by cytotoxic T cells (CTL). Another viral strategy aims to block apoptosis triggered by host defense mechanisms. Both types of strategies seem to be realized by human adenoviruses (Ads). The early transcription unit E3 of Ads encodes proteins that inhibit antigen presentation by MHC class I molecules as well as apoptosis induced by tumor necrosis factor alpha (TNF-alpha) and Fas ligand (FasL). Here, we will describe the organization of the E3 regions of different Ad subgroups and compare the structure and functions of the known immunomodulatory E3 proteins. PMID- 11022787 TI - Immune escape by hepatitis B viruses. AB - Hepatitis B viruses are DNA viruses characterized by their very small genome size and their unique replication via reverse transcription. The circular genome has been efficiently exploited, thereby limiting genome variation, and leaves no space for genes in addition to those essentially needed during the viral live cycle. Hepatitis B viruses are prototype non-cytopathic viruses causing persistent infection. Human hepatitis B virus (HBV), as well as the closely related animal viruses, most frequently are transmitted vertically from mothers to their offspring. Because infection usually persists for many years, if not lifelong, hepatitis B viruses need efficient mechanisms to hide from the immune response of the host. To escape the immune response, they exploit different strategies. Firstly, they use their structural and non-structural proteins multiplely. One of the purposes is to alter the immune response. Secondly, they replicate by establishing a pool of stable extrachromosomal transcription templates, which allow the virus to react sensitively to changes in its microenvironment by up- or downregulating gene expression. Thirdly, hepatitis B viruses replicate in the liver which is an immunopriviledged site. PMID- 11022788 TI - MHC class I-subversive gene functions of cytomegalovirus and their regulation by interferons-an intricate balance. AB - Multiple glycoproteins of human cytomegalovirus (HCMV) encoded by the genes US2, US3, US6 and US11 interrupt the MHC class I pathway of antigen presentation at distinct checkpoints to avoid recognition of infected cells by cytotoxic CD8+ T lymphocytes. The action of cytokines like interferon (IFN)-gamma, IFN-alpha/beta and tumour necrosis factor alpha (TNF-alpha) compensate for the viral inhibition and restore antigen presentation in HCMV-infected cells. This finding was explained by their effects on cellular rather than viral genes and reflected by an increase in the production, assembly and maturation of MHC class I molecules resulting in an escape of MHC I from viral control. Here we reproduce the IFN gamma-mediated effect when MHC I-subversive gene functions of HCMV are tested in isolation, but the efficacy of IFN-gamma to restore MHC I surface expression in US2-, US6- and US11-transfectants differs significantly. In addition, in HCMV infected cells IFN-gamma strongly affects the synthesis of the US6-encoded glycoprotein. Despite the capability of HCMV to block the interferon signaling pathway the IFN-gamma driven enhancement of MHC class I and class II expression remains effective provided that cells are exposed to IFN-gamma before infection. Our findings illustrate a complex interplay between host immune factors and viral immune evasion functions. PMID- 11022790 TI - Herpesvirus homologues of cellular genes. AB - For millions of years viruses have adapted strategies to interfere with the immune defense of the host, which in turn has to deal with this challenge. In general the antiviral defense remains one step behind the pathogen. To achieve this strategic advantage large DNA-containing Viruses encode cellular homologues that mimic or counteract key molecules of the host immune system. Understanding how these cellular homologues enable the viruses to evade the antiviral defense and persist in the host for the lifetime will ultimatively lead also to a better understanding of the principle functions of the immune system. In this review we focused on cellular homologues encoded by human herpesviruses and discuss the functional consequences of their expression. PMID- 11022789 TI - Marek's disease herpesvirus transforming protein MEQ: a c-Jun analogue with an alternative life style. AB - In order to adapt to and to cope with an often hostile host environment, many viruses have evolved to encode products that are homologous to cellular proteins. These proteins exploit the existing host machinery and allow viruses to readily integrate into the host functional network. As a result, viruses are able to maneuver their journey seemingly effortlessly inside the host cell to achieve ultimate survival. Such molecular mimicries sometime go overboard, allowing viruses to overtake the cellular pathways or evade the immune system as do many of the retroviral oncogenes. Retroviral oncogenes are derived directly from host genes, and they are virtually identical to host genes in sequences except those mutations that make them unregulatable by host. Oncogenic herpesviruses also encode oncogenes, or transforming genes, which have independently evolved and are distantly related to host genes. However, these genes do share consensus structural motifs with cellular genes involved in cell growth and apoptosis and are functional analogues to host genes. The Marek's disease virus oncoprotein, MEQ, is one such example. MEQ is a basic region-leucine zipper (bZIP) transactivator which shares extensive homology with the Jun/Fos family of transcription factors within the bZIP domain, but not in other regions. Like all other bZIP proteins, MEQ is capable of dimerizing with itself and with a variety of bZIP partners including c-Jun, B-Jun, c-Fos, CREB, ATF-1, ATF-2, and SNF. MEQ Jun heterodimers bind to a TRE/CRE-like sequence in the meq promoter region and have been shown to up-regulate MEQ expression in both chicken embryo fibroblasts and F9 cells. In addition, the bZIP and transactivation domains are interchangeable between MEQ and c-Jun in terms of transforming potential; i.e. MEQ can functionally substitute for c-Jun. These properties enable MEQ to engage in host cell processes by disguising itself as c-Jun. On the other hand, there are properties of MEQ notably different from c-Jun, which include its capability to bind RNA, to bind a CACAC-bent DNA structure as a homodimer, to inhibit apoptosis, and to interact with CDK2. MEQ's subcellular localization in the nucleolus and coiled body, is also different from Jun/Fos family of transactivators. These unique features may provide the MEQ with additional facility in regulating MDV replication, establishing latency, and cellular transformation. In this review, we will attempt to summarize the past research progress on MDV meq, with a focused on the similarities and differences between MEQ and cellular proteins, and between MEQ and other viral oncoproteins. PMID- 11022791 TI - Iridovirus homologues of cellular genes--implications for the molecular evolution of large DNA viruses. AB - Iridoviruses belong to the group of large cytoplasmic deoxyriboviruses and infect either insects or vertebrates. In analogy to other large DNA viruses of eucaryotes it was found that iridoviruses encode a number of cellular protein homologues. The majority of these proteins represent orthologues of cellular enzymes involved in transcription, replication, and nucleotide metabolism. Others may have the potential to interfere with cell cycle regulation or immune defence mechanisms of the host. This raises the question about the phylogenetic origin of the corresponding viral genes. During the evolution of large cytoplasmic DNA viruses such as iridoviruses, poxviruses, and African swine fever virus the acquirement of cellular genes appears to be a crucial event. Each member of this group of viruses encodes a DNA polymerase, two subunits of the DNA-dependent RNA polymerase, and two subunits of the ribonucleotide reductase. It is important to note that all of these viral proteins show a high level of multidomain structure conservation as compared to their cellular orthologues. As a consequence the large cytoplasmic DNAviruses have the ability to replicate independently of the cellular nucleus in the cytoplasm of the infected cell. Assuming a common cellular origin of viral DNA polymerase genes the corresponding amino acid sequences were chosen to construct a phylogenetic tree showing the relatedness among large DNA viruses of eucaryotes. PMID- 11022792 TI - Sequence and functional analysis of a homolog of interleukin-10 encoded by the parapoxvirus orf virus. AB - Orf virus is a large DNA virus and is the type species of the Parapoxvirus genus of the family Poxviridae. Orf virus infects the epithelium of sheep and goats and is transmissible to humans. Recently we discovered a gene in orf virus that encodes a polypeptide with remarkable homology to mammalian interleukin (IL-10) and viral encoded IL-10s of herpes viruses. The predicted polypeptide sequence shows high levels of amino acid identity to IL-10 of sheep (80%), cattle (75%), humans (67%) and mice (64%), as well as IL-10-like proteins of Epstein-Barr virus (63%) and equine herpes virus (67%). The C-terminal region, comprising two-thirds of the orf virus protein, is identical to ovine IL-10 which suggests that this gene has been captured from its host sheep during the evolution of orf virus. In contrast the N-terminal region shows little homology with cellular IL10s and in this respect resembles other viral IL-10s. IL-10 is a pleiotrophic cytokine that can exert either immunostimulatory or immunosuppressive effects on many cell types. IL-10 is a potent anti-inflammatory cytokine with inhibitory effects on non-specific immunity in particular macrophage function and Thl effector function. Our studies so far, indicate, that the functional activities of orf virus IL-10 are the same as ovine IL-10. Orf virus IL-10 stimulates mouse thymocyte proliferation and inhibits cytokine synthesis in lipopolysaccharide activated ovine macrophages, peripheral blood monocytes and keratinocytes. Infection of sheep with an IL-10 deletion mutant of orf virus has shown that interferon-gamma levels are higher in tissue infected with the mutant virus than the parent virus. The functional activities of IL-10 and our data on orf virus IL 10 suggest a role in immune evasion. PMID- 11022793 TI - Myxoma virus expresses a TNF receptor homolog with two distinct functions. AB - Myxoma virus, a member of the poxvirus family of DNA viruses, encodes many virulence factors to combat and evade the host immune responses. Among the virus encoded immuno-modulators is M-T2, a tumor necrosis factor receptor (TNF-R) homologue. M-T2 is secreted as monomeric and dimeric species that bind and inhibit rabbit TNF in a species-specific manner. Deletion analysis indicates that the anti-TNF function is mediated by the first three of four cysteine rich domains (CRDs) of M-T2. In addition, the intracellular form of M-T2 has the ability to block virus-induced apoptosis in lymphocytes, and the first two CRDs appear to be sufficient for this function. Although the mechanisms for the anti TNF and anti-apoptotic functions of M-T2 are not yet fully defined, we postulate that these dual activities of M-T2 are mediated through different functional motifs and abrogate distinct cellular responses to virus infection. PMID- 11022795 TI - Clinical use of cefuroxime in paediatric community-acquired pneumonia. AB - Cefuroxime has been recommended as a component of treatment for community acquired pneumonia (CAP) in guidelines produced by several groups, including the US and British Thoracic Societies. It is effective in vitro against the major bacterial pathogens in CAP but it needs to be given with an agent that is active against Mycoplasma, Chlamydia or Legionella spp. if the presence of any of these organisms is suspected. Cefuroxime penetrates respiratory tissue effectively after either parenteral or oral administration, and it has a pharmacodynamic profile which suggests that adequate cover can be achieved with oral therapy for respiratory pathogens susceptible to cefuroxime concentrations of 4 mg/L or less. This break-point is applicable to oral monotherapy and to sequential therapy regimens for the treatment of pneumonia. Cefuroxime can be used either orally or parenterally and it is approved in many countries for the treatment of adult pneumonia by either route. The oral form, cefuroxime axetil, has been used extensively in the treatment of children aged over 3 months but its use in paediatric pneumonia has not been reviewed. The present review summarises clinical experience in the treatment of bacterial pneumonia, of varying severity, in children. The data show that children with severe pneumonia, including those with pleural effusion or complications, can be treated with a full course of intravenous cefuroxime therapy, whereas hospitalised children whose pneumonia stabilises rapidly after initial intravenous therapy can change to oral cefuroxime axetil after 24 to 72 hours and may be able to return home. Oral cefuroxime axetil was appropriate for patients with milder pneumonia managed either in hospital or at home. PMID- 11022796 TI - Diamond-Blackfan Anaemia: an overview. AB - Diamond Blackfan Anaemia (DBA) is a congenital disease characterised by defective erythroid progenitor maturation. It is usually diagnosed during the first year of life. The main clinical sign is profound isolated normochromic or macrocytic anaemia, with normal numbers and function of the other haemopoietic cells. Reticulocyte counts in patients with DBA are very low. Bone marrow reflects the defective erythropoiesis, showing a very low number of erythropoietic precursors and a reduction of erythroid burst-forming unit progenitor cells. The proliferation and differentiation of the other lineages are normal. More than one third of patients have malformations, most often involving the upper limbs and head, and the urogenital or cardiovascular systems. However, the link between these malformations and defective erythropoiesis is unclear and a defect in a molecule acting on both early embryonic development and haematopoiesis has been proposed. Whereas most cases are sporadic, inheritance is observed in 10% of patients, with a dominant or, more rarely, recessive pattern. One locus on chromosome 19q13.2 encoding ribosomal protein S19 accounts for a quarter of patients with either the dominant or the sporadic form. Families not linked with this locus have also been described. The diagnosis of DBA may be difficult and differential diagnoses include Fanconi's anaemia and acquired erythroid aplasias. Erythrocyte adenosine deaminase levels are generally high in DBA patients, which may help in the diagnosis, but they are not pathognomic. Corticosteroids are the main treatment option in DBA and these agents induce erythropoiesis in over 60% of patients. Some patients achieve complete remission, which may be either corticosteroid-induced or spontaneous. The increased in vitro erythropoiesis occasionally induced by the addition of specific cytokines, namely interleukin (IL)-3 and stem cell factor (SCF), has suggested their use in vivo. However, few patients have responded to IL-3, whereas SCF administration, though interesting in theory, has not yet been attempted. Patients who do not respond to corticosteroids and those who have to discontinue treatment because of adverse events must rely on long term transfusions, and are thus exposed to all of the associated complications. Bone marrow or cord blood transplantation has been performed in some patients. The former approach is burdened with severe complications and high mortality. PMID- 11022797 TI - Helicobacter pylori infection and eradication in paediatric patients. AB - Helicobacter pylori is now recognised to be typically acquired during childhood. Studies also indicate that the infection is frequently lost in childhood; however, it is still unclear whether this is related to the use of antibacterials, the natural history of the infection, or both. H. pylori colonises gastric mucosa and is causally related to chronic gastritis and peptic ulcer disease in both children and adults. Successful eradication of H. pylori has resulted in the healing of duodenal ulcers and the lowering of the ulcer relapse rate in children. Therapy to cure the infection should be started in all children with peptic (duodenal or gastric) ulcer who are still infected. The ideal anti-H. pylori regimen should be safe, cheap, easy to comply with, well tolerated by children and able to achieve a high cure rate. Although US data are lacking, it is anticipated that the treatment regimen for children should be similar to that in adults (a triple therapy regimen that combines a proton pump inhibitor with 2 antimicrobial agents for 14 days). It is inappropriate to prescribe anti-H. pylori therapy without a firm diagnosis. The use of multiple antibacterials in a paediatric patient with an ulcer but without H. pylori infection cannot provide any benefit to the patient or the community. Such an approach only provides the possibility for adverse effects, for example development of antibacterial resistance among bystander bacteria. It is very important to confirm the diagnosis of H. pylori infection. The [13C]urea breath test is the noninvasive method of choice to determine H. pylori status in children and the ideal test for post-therapy testing. There is a need for post therapy confirmation because of the likelihood of poor outcome for some treatment regimens, which is why post-therapy testing should be the standard of care. There is weak and inconsistent evidence of an association between H. pylori infection and recurrent abdominal pain (RAP) in children, in part because of the unclear definition of RAP in the literature. Therefore, there is still considerable debate regarding the treatment of infected children with RAP. PMID- 11022794 TI - Poxvirus homologues of cellular genes. AB - Over the course of time poxviruses have acquired or "captured" numerous homologues of cellular genes and incorporated them into their large DNA genomes. With more poxvirus genome sequencing data becoming available, the number of newly discovered poxviral cellular homologues is constantly increasing. A common feature of these genes is that they are nonessential for virus replication in vitro and they confer selective advantages in dealing with host cell differentiation and immune defense mechanisms in vivo. Poxviral cellular homologues are reviewed in this synopsis considering the specific viral habitats of different poxviruses and the immune defence capabilities of their respective hosts. Possible mechanisms of cellular gene acquisition by poxviruses as suggested by the analysis of mobile genetic elements in large DNA viruses are discussed. The investigation of poxvirus homologues of cellular genes is essential for our understanding of the mechanisms that regulate virus/host interactions on the cellular level and the host response against infection. PMID- 11022798 TI - Antileukotriene drugs in childhood asthma: what is their place in therapy? AB - Leukotriene antagonists are a new class of anti-inflammatory drugs which have shown clinical efficacy in the management of asthma. However, their role in paediatric asthma is still unclear. In essence, while there are theoretical reasons as to why leukotriene antagonists would be of use in the management of childhood asthma, there is little clinical data on their use in this patient group. Studies with leukotriene antagonists to date have been performed in children with chronic 'undertreated' asthma which, under current recommendations, should be treated with inhaled corticosteroids. Furthermore, the magnitude of effect of leukotriene antagonists on lung function (forced expiratory volume in one second of less than 5% better than placebo) and daily symptoms (0.23 puffs per day of B2 agonist less than placebo), while reaching statistical significance, is unlikely to be of clinical significance in children with chronic undertreated asthma. There is, however, good evidence for leukotriene antagonist use in exercise induced asthma in children. We conclude that although leukotriene antagonists may play an important role in the management of childhood asthma in the future, particularly as corticosteroid sparing agents and in exercise induced asthma, clinical data in paediatric asthma is poor. PMID- 11022800 TI - The link between family history and early onset alcoholism: earlier initiation of drinking or more rapid development of dependence? AB - OBJECTIVE: This study examines the association between early onset alcoholism and family history to determine whether family history of alcoholism is predictive of earlier initiation of drinking, more rapid onset of dependence once drinking has begun, or both. METHOD: Using cross-sectional, retrospective data from a large, nationally representative sample of U.S. adults, discrete time proportional hazards models were used to assess the effects of family history saturation (% of alcoholic first- and second-degree relatives) on: ( 1) the risk of initiating drinking among all adults (N = 42,862; 58.4% female) and (2) the risk of progressing from initiation of drinking to onset of dependence among lifetime drinkers (n = 27,616; 50.7% male). Models were estimated for different time periods, to see if the effect of family history saturation varied over time in a manner suggestive of a stronger association with early onset dependence. RESULTS: The positive effect of family history saturation on the risk of initiating drinking was strongest prior to age 15 and declined steadily with increasing age. It was slightly weaker for men than women. After controlling for early initiation of drinking, the direct positive effect of family history saturation on the risk of progressing to dependence increased over time and was slightly reduced among individuals who started drinking before age 18. The indirect effect of family history on the risk of developing dependence, via its effect on early drinking as a risk factor for dependence, was strongest in the interval from 3 to 9 years after initiation of drinking. CONCLUSIONS: The association between family history and early onset alcoholism appears to be driven most clearly by family history predicting earlier initiation of drinking. The weak effect of family history on the development of dependence within the first 3 years after initiation of drinking may reflect the preponderance of developmentally limited dependence during this time period. The data are consistent with the links established between novelty seeking, impulsivity and early onset alcoholism. While supporting the possibility of genetic effects via dopaminergic and serotonergic function, these findings also suggest that environmental factors may play an important part in helping to explain the association between family history and early onset alcoholism. PMID- 11022801 TI - Acute tolerance, alcohol sensitivity and drinking pattern in the F2 generation of UChA and UChB rats. AB - OBJECTIVE: Studies in mice and rats bred for their behavioral response to ethanol have revealed that there may be a relationship between acute tolerance development, sensitivity to ethanol and alcohol preference. We saw a need to obtain more genetically meaningful correlations between ethanol consumption and acute tolerance to motor impairment or sensitivity to hypothermia induced by ethanol. In the present article, we provide results on acute tolerance, sensitivity to hypothermia and voluntary ethanol consumption in an F2 generation of a cross between inbred UChA (low) and UChB (high) ethanol-consuming rats. METHOD: Naive UChA and UChB rats were tested for acute tolerance development to motor impairment and sensitivity to hypothermia induced by the intraperitoneal administration of a dose of ethanol of 2.3 g/kg body weight. Rats were then offered ad libitum a 10% v/v ethanol solution, distilled water and rat food, and classified according to their drinking scores. Next, one female UChA (low consumer) was mated with one male UChB (high consumer) and one female UChB (high consumer) mated with one male UChA (low consumer). The F2 generation of these inbred rats was used to test acute tolerance, sensitivity and voluntary ethanol consumption. RESULTS: UChB rats developed acute tolerance more rapidly and were less sensitive to the ethanol dose than UChA rats. The F2 generation was designated Hybrid A (HA) for those offspring from the UChA female grandparent and Hybrid B (HB) for the offspring from the UChB female. Results show clearly that there is a relationship between the ability of the rat to acquire acute tolerance to motor impairment, the sensitivity to hypothermia induced by ethanol, and the ethanol preference of the rat. The multiple correlation analysis between the three behaviors showed a significant value in HA rats (r = -0.76, p < .01) and in HB rats (r = -0.83, p < .01). CONCLUSIONS: This study of a cross between UChA and UChB rats showed that the three parameters are related since they clustered together in the F2 generation. This result leads us to speculate that if we could control acute tolerance development in the rats we might also control their voluntary ethanol consumption. PMID- 11022799 TI - Nevirapine: a review of its use in the prevention and treatment of paediatric HIV infection. AB - Nevirapine is a highly specific inhibitor of HIV-1 reverse transcriptase (RT), an important therapeutic target for the treatment of HIV infection. It was the first non-nucleoside RT inhibitor (NNRTI) to be approved for use in HIV-infected individuals, including children. Nevirapine inhibits replication of multiple HIV 1 strains and clinical isolates in cultured human T cells, but has no activity against other retroviral RTs (including HIV-2 RT) or endogenous human DNA polymerases. Monotherapy with nevirapine rapidly selects for high level drug resistance conferred by a single amino acid substitution in the HIV RT gene. The pattern of resistance mutations selected by nevirapine overlaps with those of other NNRTIs, but is distinct from those of nucleoside analogue RT inhibitors and protease inhibitors. The pharmacokinetics of nevirapine are characterised by rapid and nearly complete oral absorption, an apparently even distribution throughout all organs and tissues in the body, and a long elimination half-life. Nevirapine is both metabolised by and induces the activity of cytochrome P450 isoenzymes. Caution is required when coadministering nevirapine with other drugs metabolised by this system, including HIV protease inhibitors. As a single dose therapy given to pregnant women in labour and to neonates shortly after birth, nevirapine reduced the rate of perinatal HIV transmission by 47% compared with a short course of intrapartum and neonatal zidovudine in a randomised trial in breastfeeding women in Uganda. Nevirapine was more cost effective than zidovudine based regimens for the prevention of perinatal HIV transmission in a cost effectiveness model in sub-Saharan Africa. Nevirapine has shown antiretroviral efficacy as part of combination therapy with zidovudine and either didanosine or lamivudine in small numbers of paediatric patients in phase I/II trials. Triple therapy of nevirapine, zidovudine and didanosine was moderately effective in a randomised, nonblind trial in patients with advanced disease. The most frequent adverse event associated with nevirapine is rash, which occasionally develops into severe rash or Stevens-Johnson syndrome. Elevations in liver enzyme levels can occasionally lead to severe complications in patients receiving multiple dose nevirapine. CONCLUSION: Single dose nevirapine appears to be the most cost effective of available options for the prevention of perinatal HIV transmission suitable for use in developing countries. In addition, preliminary clinical trial data suggest that nevirapine adds to the efficacy of combination therapy for the treatment of HIV infection in infants and children, and is reasonably well tolerated. PMID- 11022802 TI - A moderate intake of wine is associated with reduced total mortality and reduced mortality from cardiovascular disease. AB - OBJECTIVE: The aim of this study was to investigate the effects of the consumption of wine, beer and distilled spirits on total mortality and on mortality from cardiovascular disease. METHOD: The consumption of wine, beer and distilled spirits was assessed in 1,828 individuals by a psychiatrist. Subjects were selected according to expected level of need for health services, from a random sample of 24,043 individuals aged 18-65 years. Mortality was recorded after 22 years and the results related to those for the individuals not exposed to the factor examined. The results were adjusted for age, expected level of need for health services, total alcohol consumption, gender, body-mass index, tobacco use and social class. RESULTS: Intake of wine once a week or more (compared with intake of wine less than once a week or not at all) was associated with a relative risk ratio of 0.58 for total mortality (95% CI: 0.40-0.84) and a relative risk ratio of 0.49 for mortality from cardiovascular disease (95% CI: 0.27-0.90). The risk reduction seemed to be confined to those consumers of wine who had an intake of less than 140 grams of alcohol per week and consumed the beverage once a week. Ex-drinkers had an increased relative risk ratio in total mortality compared with lifelong abstainers and individuals who consumed less than 50 grams of alcohol per week (relative risk ratio = 2.64; 95% CI: 1.56 4.49). CONCLUSIONS: A low to moderate intake of wine seems, unlike the consumption of distilled spirits and beer, to be associated with reduced total mortality and reduced mortality from cardiovascular disease. PMID- 11022803 TI - Methylphenidate treatment of neuropsychiatric symptoms of central and extrapontine myelinolysis. AB - OBJECTIVE: Previous reports describe the presentation and course of the neurobehavioral manifestations of central and extrapontine myelinolysis; as of yet, however, there are no specific recommendations for treatment of these problems. We offer the first report of successful treatment. METHOD: We describe a 55-year-old man with chronic alcoholism who developed central and extrapontine myelinolysis following an episode of heavy drinking and rapid correction of hyponatremia. The patient acutely developed motor, cognitive, emotional and behavioral problems best accounted for by central pontine and bilateral striatal myelinolysis. These neuropsychiatric symptoms were treated with methylphenidate over the course of 1 month in an off-on-off-on fashion. The Neuropsychiatric Inventory and other tests were used to assess treatment response. RESULTS: Marked improvements in the patient's neuropsychiatric status were noted only during treatment with methylphenidate. CONCLUSIONS: Methylphenidate effectively reversed the neuropsychiatric symptoms associated with the patient's demyelinating lesions. We discuss possible underlying mechanisms of both symptom formation and treatment effect. PMID- 11022805 TI - Peer networks among heavy, regular and infrequent drinkers prior to marriage. AB - OBJECTIVE: Research has consistently demonstrated that, among adolescents, the characteristics of one's peers are important predictors of substance abuse. The impact of the peer network on adult drinking, however, has received considerably less attention. The purpose of the present study was to examine social network characteristics that are associated with heavy drinking in adulthood prior to marriage. METHOD: Couples were recruited at the time of their first marriage. Husbands and wives were each given identical questionnaire packets to complete at home, independently, as well as a postage-paid envelope for packet return. A broad range of constructs was assessed; included were personality characteristics, relationship functioning, drinking behavior and social network characteristics. Complete data were obtained from 471 husbands and 471 wives. RESULTS: The social networks of heavy-drinking men, compared to men drinking regularly or infrequently, were younger, more likely to be male and unmarried and consisted of friends rather than family or others. For both men and women, "drinking buddies" accounted for nearly 75% of the heavy drinkers' peer networks. The overall ratings of support and conflict created by peers did not differ according to drinking group, for either men or women. CONCLUSIONS: Prior to marriage, the social networks of heavy drinkers differ considerably from the networks of regular or infrequent drinkers with regard to the drinking patterns of their peers. An important finding was that heavy drinkers appear to experience a similar level of emotional, financial and practical support from their peer network compared to regular or infrequent drinkers. PMID- 11022804 TI - Maternal smoking and drinking during pregnancy and the risk for child and adolescent psychiatric disorders. AB - OBJECTIVE: To examine the relative importance of prenatal exposure to cigarettes and alcohol and familial/genetic susceptibility for alcohol dependence in the etiology of childhood psychopathology. METHOD: A longitudinal prospective study of 150 children/adolescents (51.3% male), who were at either high or low risk for developing alcohol dependence because of their familial loading for alcoholism, provided multiple diagnostic assessments (N = 318) of these subjects. High-risk families were identified through the presence of two adult alcoholic sisters; low risk control families were selected from the community. Annual assessments of offspring from these families included an in-depth psychiatric interview of each child and his/her parent to determine the presence or absence of childhood disorders. Mothers were interviewed concerning their prenatal use of substances, and information was gathered concerning their personal and familial loading for psychiatric disorders. RESULTS: Using conventional logistic regression analyses, internalizing and externalizing disorders were found to be associated with familial loading for alcoholism and prenatal exposure to cigarettes and alcohol. In addition, a specialized statistical analysis, a multivariate confounder score approach, was conducted using familial risk status and the child's exposure to maternal prenatal use of alcohol and cigarettes. This analysis demonstrated that only one relationship between a single variable and a childhood disorder was significant while controlling for the other two variables: Oppositional disorder remained significant in association with familial risk status. Three additional analyses were performed to evaluate the effects of familial risk status, prenatal alcohol exposure and prenatal cigarette exposure on childhood psychopathology while controlling for two known risk factors (SES and parental ASPD) for externalizing disorders. Results of these analyses revealed that the only childhood disorder that was elevated was ADHD, and that this was the result of the familial risk variable only. CONCLUSIONS: Familial loading for alcohol dependence is an important risk factor for the development of childhood psychopathology and may account for the previously reported associations between prenatal exposure to nicotine and alcohol. Studies of substance abuse/dependence etiology and childhood psychopathology need to include consideration of both prenatal exposures and familial loading for alcohol dependence and other psychiatric disorders. PMID- 11022806 TI - Readiness to change drinking among heavy-drinking college students. AB - OBJECTIVE: The purpose of this study was to examine motivation to reduce alcohol consumption among heavy-drinking college students. Specific goals were to test the factor structure of the Stages of Change Readiness and Treatment Eagerness Scale (SOCRATES); present normative SOCRATES data for collegiate heavy drinkers; present a heuristic, using SOCRATES scales to determine stage of readiness to change heavy-drinking; and compare students at different stages of change on demographic and drinking variables. METHOD: Participants were 278 (187 female) undergraduates who reported at least one episode of heavy drinking within the past 3 months. Students completed the SOCRATES and other questionnaires that assessed current and past drinking and demographics. RESULTS: Confirmatory factor analysis provided modest support for the SOCRATES factor structure. Students were classified according to the transtheoretical model of change (67% Precontemplation, 20% Contemplation and 13% Action). Contemplators drank more often, consumed more alcohol, reported more heavy drinking episodes and experienced more alcohol consequences than the other groups. CONCLUSIONS: Two thirds of the heavy-drinking college students did not recognize a need to reduce their alcohol consumption, despite evidence of tolerance and negative drinking consequences. PMID- 11022807 TI - Expectations of alcohol-induced positive affect: a cross-cultural comparison. AB - OBJECTIVE: Our aim was to investigate the extent to which positive affect is a cross-culturally expected result of drinking alcohol. This first required development of a quantitative estimate of positive affect on a common scale, an essential step neglected in previous comparative research on alcohol expectancies. METHOD: Approximately equal numbers of male and female respondents (N = 1,008; 521 women) from eight countries were asked to complete a survey inquiring about emotional and behavioral responses they expect people to experience after consuming alcoholic drinks, and about limited aspects of their own drinking habits. Multisample latent covariance structure analysis with means was applied to the data and a cross-culturally invariant model of positive affect was extracted. RESULTS: The latent construct for positive affect that emerged was defined by manifest "interpersonal warmth or closeness," "pleasure of social interactions" and "optimism." There were significant national differences in means for this factor, and self-reported drinking frequency was also marginally related to expected positive affect. CONCLUSIONS: Multisample latent covariance structure analysis with means proved a useful tool capable of addressing critical problems in comparative cross-cultural research. In addition, there were indications that the expectation of increased positive affect associated with drinking may be influenced by contextual factors and cultural traditions, making positive affect less easily attributable to the direct pharmacological action of alcohol consumption than has previously been believed. PMID- 11022808 TI - Alcohol's effects on sexual perception. AB - OBJECTIVE: This study examines hypotheses about alcohol's effects on sexual judgments based on both alcohol and misperception theories. It was hypothesized that gender, alcohol consumption and alcohol expectancy set would influence perceptions of sexuality. METHOD: Participants were unacquainted women and men (88 dyads) who interacted for 15 minutes within the context of the balanced placebo design. After the conversation ended, participants answered questions about their behavior and their partners' behavior. Conversations were videotaped and coded by trained raters. RESULTS: Men perceived their female partner and themselves as behaving more sexually than women perceived their male partner and themselves. When alcohol was consumed, both women and men were perceived as behaving more sexually and in a more disinhibited manner than when alcohol was not consumed. Ratings made by members of white and black dyads were largely comparable. Trained observers coded participants' use of active attention and dating availability cues. Both types of cues interacted with alcohol consumption such that intoxicated participants exaggerated the meaning of strong (dating availability) cues and ignored the meaning of ambiguous (active attention) cues when making sexual judgments. CONCLUSIONS: Supporting past research on gender differences in perceptions of sexuality, men were more sexually attracted to their opposite-sex partner than women were. Both women's and men's sexual judgments were influenced by alcohol consumption but not by alcohol expectancy set. Intoxicated participants' responses to their partners' behavioral cues supported cognitive impairment models of alcohol's effects. The implications of these findings for theories about alcohol's effects on sexuality and for prevention programming are discussed. PMID- 11022809 TI - Variables associated with DUI offender status among whites and Mexican Americans. AB - OBJECTIVE: Arrests of Hispanics have been found to be overrepresented among arrests for driving under the influence of alcohol (DUI) in California. Variables associated with offender status (first vs multiple) among Mexican-American and white DUI arrestees are analyzed to test the hypothesis that acculturation will be related to DUI among Mexican Americans while risk-taking dispositional variables will be related to DUI among whites. METHOD: A sample of 249 Mexican American clients (with an oversampling of foreign-born) and 250 white clients were interviewed in all five DUI treatment programs in one northern California county. RESULTS: In univariate analysis, risk taking/impulsivity and sensation seeking were positively associated with multiple offender status among Mexican Americans, but not among whites. Multiple logistic regression found age as the only significant predictor of offender status in either ethnic group; however, path models found significant indirect effects on offender status of attitudes regarding DUI arrest through alcohol consumption, with those reporting having little chance of being arrested for DUI if stopped being less likely to report consuming five or more drinks at a sitting at least weekly, and with Mexican Americans being more likely than whites to believe they will not be arrested and to consider that DUI is not a problem. CONCLUSIONS: The data suggest that there are significant ethnic differences in attitudes and norms toward DUI that are related to frequency of drinking large amounts at one sitting, and future research is needed to determine how these variables along with dispositional and acculturation variables may be associated with DUI offender status. PMID- 11022810 TI - Specialty mental health care improves patients' outcomes: findings from a nationwide program to monitor the quality of care for patients with substance use disorders. AB - OBJECTIVE: To describe the implementation of a nationwide program to monitor the quality of treatment for substance use disorders in the Department of Veterans Affairs, and to examine how the provision of outpatient mental health care, and the duration and intensity of care, relate to patients' outcomes. METHOD: Clinicians completed a baseline Addiction Severity Index (ASI) on more than 34,000 patients with substance use disorders; more than 21,000 (63%) were reassessed with the ASI an average of 12 months later. Nationwide health service utilization databases were used to obtain information about patients' diagnoses and their use of services during an index episode of care. RESULTS: On average, patients who received specialty outpatient mental health care experienced better risk-adjusted outcomes than did patients who did not receive such care. Patients who had longer index episodes of mental health care improved more than did those who had shorter episodes. There was some evidence that the duration of care contributed more to better outcomes among patients with only substance use disorders, whereas the intensity of care was more important for patients with both substance use and psychiatric disorders. CONCLUSIONS: The provision of specialty outpatient mental health care, and longer episodes of specialty care, were associated with better risk-adjusted substance use, symptom and social functioning outcomes for patients with substance use disorders. More emphasis should be placed on ensuring that these patients enter specialty care and on keeping them in treatment. PMID- 11022811 TI - Predictors of self-help group attendance in cocaine dependent patients. AB - OBJECTIVE: While referral to self-help groups for patients dependent on drugs other than alcohol has become widespread in the substance abuse treatment field, little is known about the characteristics of people who attend these groups. This study examines particular sociodemographic and clinical characteristics as possible predictors of attendance at self-help groups in the National Institute on Drug Abuse Collaborative Cocaine Treatment Study. METHOD: A multicenter study randomly assigned 487 patients (76.8% men) to one of four psychosocial treatments for cocaine dependence. Patients were treated for 24 weeks. Among other measures, the Weekly Self-Help Questionnaire was administered each week and completed, at least once, by 411 subjects. RESULTS: Approximately two thirds of the patients attended one or more self-help meetings during the 24-week period. Patients initially more likely to attend self-help groups frequently were those who were unemployed, had no religious preference, had more severe baseline drug use and reported treatment for prior substance-related problems. Patients with more severe baseline drug use and those who previously received treatment for substance-related problems were more likely to maintain frequent attendance throughout the study period. Only severity of baseline drug use predicted more frequent attendance during Month 6, although there was a trend in Month 6 favoring more frequent attendance by women. These findings remained significant when treatment condition was added to the models. CONCLUSIONS: Although these findings are consistent with past research on alcohol dependent individuals, they challenge popular clinical notions about the types of people who attend self-help groups. These findings demonstrate that self-help groups can appeal to a wide variety of cocaine dependent patients. PMID- 11022812 TI - Comparison of outcomes for clients in randomized versus open trials of treatment for alcohol use disorders. AB - OBJECTIVE: There has been continuing concern that clients who accept randomization into a controlled trial may not be representative of those in noncontrolled trials or ordinary treatment situations. However, it is not possible to test the impact of randomization through a randomized trial. Two parallel studies conducted at the same treatment facility provided an opportunity for a quasi-experimental study to evaluate whether participation in a controlled trial itself affects treatment outcome. METHOD: Two concomitant samples of clients were enrolled during overlapping recruitment periods: one (n = 226) into a randomized clinical trial (RCT) and the other (n = 122) offered treatment as usual (non-RCT). Both samples were given extensive baseline and follow-up assessment. RESULTS: Baseline assessment indicated demographic similarity of the two samples, with somewhat higher problem severity in the non-RCT sample, consistent with the RCT selection criteria. Client retention in treatment was somewhat comparable, and follow-up rates exceeded 90% in both studies. Overall outcomes did not differ for the RCT and non-RCT samples. CONCLUSIONS: It appeared that clients enrolled into an RCT did not differ from those receiving ordinary treatment. Retention was similarly high in both studies, clients completed a comparable number of outpatient sessions, and the number of informal treatment sessions attended during the 6 months of follow-up was comparable. There are some aspects of this study that limit the ability to draw firmer conclusions but despite some pretreatment differences, participation in the RCT did not itself exert an apparent effect on aggregate treatment outcomes. PMID- 11022813 TI - Clinical characteristics and family histories of alcoholics with stimulant dependence. AB - OBJECTIVE: While much is known about the clinical patterns and family histories of individuals with alcoholism or stimulant (cocaine and amphetamine) dependence, there are few data that describe men and women with concomitant alcohol and stimulant dependence. METHOD: As part of the Collaborative Study on the Genetics of Alcoholism, structured interviews were administered to 3,882 (2,432 male) DSM III-R defined alcohol and/or stimulant dependent subjects. The characteristics and family histories of four groups were compared: Group 1 (26%), with the onset of alcohol before stimulant dependence; Group 2 (10%), with alcohol dependence simultaneously with or after stimulant dependence; Group 3 (58%), with alcohol dependence only; Group 4 (6%), with stimulant dependence only. RESULTS: Individuals with concomitant alcohol and stimulant dependence (Groups 1 and 2) reported more general life problems (e.g., marital instability), a higher rate of antisocial personality disorder and more substance-induced mood disorders, additional drug dependencies and substance-related difficulties than those with dependence on one substance only. People with alcohol dependence before stimulant dependence had the most severe clinical patterns. In addition, alcohol dependence and stimulant dependence were found to breed true in families of subjects with these concomitant disorders. The major findings were confirmed with logistic regression analyses, and were independent of ASPD and gender. CONCLUSIONS: It is important for clinicians to be aware of the severe clinical characteristics of patients with concomitant alcohol and stimulant dependence. In addition, the data consistent with drug-specific heritability in this heterogeneous population may be useful to researchers. PMID- 11022814 TI - Alcohol treatment research follow-up interviews and drinking behaviors. AB - OBJECTIVE: This study, drawing upon data collected as part of a randomized clinical trial of alcohol treatment matching effects, investigates the relationship between research follow-up assessment interviews and subsequent drinking behaviors. METHOD: Subjects (N = 188; 143 men) participated in a day hospital substance-use disorder treatment program at either a private psychiatric hospital (n = 151) or a community hospital (n = 37) and were classified into one of three research groups: regularly scheduled follow-up interviews, missed scheduled follow-up interviews, and delayed Year-2 follow-up interviews. Complete data relevant to the present study were collected on 157 subjects. RESULTS: Study results provided support for a subject reactivity effect related to the research follow-up interview; that is, those subjects classified within the regularly scheduled follow-up interviews condition had the better drinking outcomes. Furthermore, by the end of the second follow-up year, subjects classified within the delayed Year-2 follow-up interviews condition had the poorest drinking outcomes. CONCLUSIONS: There may be considerable potential for subject reactivity effects, specific to the research assessment interview, to confound study results. Therefore, the interpretation of data from alcohol treatment outcome studies that fail to control for such potential confounding effects must be viewed with caution. Researchers are advised to control for these potentially confounding effects via methodological and/or statistical mechanisms. PMID- 11022815 TI - An examination of the utility of the AUDIT in people with schizophrenia. AB - OBJECTIVE: To examine the reliability and validity of the Alcohol Use Disorders Identification Test (AUDIT) compared to a structured diagnostic interview, the Composite International Diagnostic Interview (CIDI; 12-month version) in psychiatric patients with a diagnosis of schizophrenia. METHOD: Patients (N = 71, 53 men) were interviewed using the CIDI (Alcohol Misuse Section; 12-month version) and then completed the AUDIT. RESULTS: The CIDI identified 32.4% of the sample as having an alcohol use disorder. Of these, 5 (7.0%) met diagnostic criteria for harmful use of alcohol, 1 (1.4%) met diagnostic criteria for alcohol abuse and 17 (23.9%) met diagnostic criteria for alcohol dependence. The AUDIT was found to have good internal reliability (coefficient = 0.85). An AUDIT cutoff of > or = 8 had a sensitivity of 87% and specificity of 90% in detecting CIDI diagnosed alcohol disorders. All items except Item 9 contributed significantly to discriminant validity. CONCLUSIONS: The findings replicate and extend previous findings of high rates of alcohol use disorders in people with severe mental illness. The AUDIT was found to be reliable and valid in this sample and can be used with confidence as a screening instrument for alcohol use disorders in people with schizophrenia. PMID- 11022816 TI - Factor structure of the Alcohol Use Disorders Identification Test (AUDIT) in a mental health clinic sample. AB - OBJECTIVE: To examine the factor structure of the Alcohol Use Disorders Identification Test (AUDIT) and to identify the implications of this structure for its clinical use. METHOD: The AUDIT was administered to mental health clinic outpatients (N = 197; 86% men) at high risk for alcohol-use disorders. Confirmatory and exploratory factor analyses were used to determine the underlying factor structure of the AUDIT for this high-risk population. RESULTS: Confirmatory analyses indicated that the a priori three- and one-factor solutions did not fit the observed data. The exploratory analyses supported a two-factor solution that included level of alcohol consumption and drinking problems, with both factors explaining substantial variance in AUDIT scores. These findings contrast the original three-factor design of the AUDIT and the conventional use of the AUDIT as a one-factor screening device with a single cutoff score. CONCLUSIONS: Other screening methods that incorporate this two-factor model may be important for mental health patient populations. Replication of these findings among other mental health samples is needed. PMID- 11022817 TI - A simple scale of Gorski's warning signs for relapse. AB - OBJECTIVE: Though it has enjoyed widespread popularity, Gorski's post-acute withdrawal syndrome (PAWS) model of relapse has been subjected to little scientific scrutiny. A scale to operationalize Gorski's 37 warning signs was developed and tested in a larger prospective study of predictors of relapse. Of central interest were: (1) whether the warning signs hypothesized by Gorski are interrelated in a meaningful single factor and (2) whether the hypothesized syndrome would accurately predict subsequent relapses. METHOD: A sample of 122 individuals (84 men) entering treatment for alcohol problems was followed at 2 month intervals for 1 year. The Assessment of Warning-signs of Relapse (AWARE) scale was administered at each assessment point, and the occurrence of both slips (any drinking) and relapses (heavy drinking) was monitored during each subsequent 2-month interval. Principal factor analysis was used to study the factor structure of the warning signs. RESULTS: Of the 37 warning signs, 28 clustered as a robust single factor with excellent internal consistency (Cronbach's alpha: 0.92-0.93). A conservative evaluation of test-retest stability across 2-month intervals estimated reliability at r = 0.80. After covarying for prior drinking status, clients' AWARE scores significantly predicted subsequent slips and relapses. Relapse rates for clients with highest AWARE scores, as projected by regression equations, were 33 to 46 percentage points higher than those for clients with lowest AWARE scores, after taking into account prior drinking status. CONCLUSIONS: This scale of Gorski's warning signs appears to be a reliable and valid predictor of alcohol relapses. PMID- 11022818 TI - Temptation, restriction, and the regulation of alcohol intake: validity and utility of the temptation and restraint inventory. AB - OBJECTIVE: The Temptation and Restraint Inventory (TRI) is a measure of drinking restraint, the preoccupation with controlling alcohol intake. It contains two higher order factors pertaining to the temptation to drink and the restriction of alcohol intake. We tested the convergent and discriminant validity of the TRI in comparison to the Preoccupation with Alcohol Scale (PAS), the Reasons for Limiting Drinking Scale (RLD) and the Drinking-Related Locus of Control Scale (DRIE), each of which measures constructs analogous to the components of the TRI. We also tested the utility of the TRI, as compared to these related measures, for predicting typical drinking and alcohol problems. METHOD: A sample of 296 (50% male) moderate to heavy drinking adults completed the TRI, the PAS, the RLD, the DRIE and measures of typical (weekly) drinking and alcohol problems (SMAST). RESULTS: Correlations indicated that the TRI had excellent convergent and discriminant validity. The TRI's temptation factor (Cognitive and Emotional Preoccupation) was strongly and positively correlated with the PAS and the DRIE, and was weakly and positively correlated with the RLD. The TRI's restriction factor (Cognitive and Behavioral Control) was positively and moderately correlated with the DRIE and moderately to weakly correlated with the PAS and the RLD. In regressions predicting weekly drinking and SMAST scores, the TRI was particularly effective in accounting for the variance in SMAST scores. CONCLUSIONS: In comparison to measures of conceptually related constructs, the components of the TRI showed excellent convergent and discriminant validity. The TRI accounted for large portions of the variance in alcohol-related outcomes, using fewer items than measures of similar constructs. PMID- 11022819 TI - JGH and Asia-Pacific consensus on prevention and management of gastrointestinal and liver diseases. PMID- 11022820 TI - Biochemical markers of hepatic fibrogenesis: single measurements are not reliable enough to replace liver biopsy. PMID- 11022821 TI - Treatment of hepatocellular carcinoma. PMID- 11022822 TI - Consensus statements on the prevention and management of hepatitis B and hepatitis C in the Asia-Pacific region. Core Working Party for Asia-Pacific Consensus on Hepatitis B and C. PMID- 11022823 TI - Oesophageal varices: assessment of the risk of bleeding and mortality. AB - Patients with oesophageal varices run a high risk of bleeding and even death, however rates of bleeding and mortality vary greatly. Indeed, a number of patients with varices never bleed. Prophylactic therapy is effective, but can be associated with side-effects. It remains to be determined which patients are at high risk of bleeding and require treatment. In addition, since non-response to medical therapy has been reported to occur in 20-40% of patients, the effect of a given prophylactic drug, or combinations of drugs, needs to be tested. A review is given of available methods of assessment. The Hepatic Venous Pressure Gradient, and measurements of the variceal pressure, are two proven methods, and the latter has the advantages of being non-invasive and having value in presinusoidal portal hypertension. PMID- 11022824 TI - Effect of branched-chain amino acids on the composition and cytolytic activity of liver-associated lymphocytes in rats. AB - BACKGROUND: Although branched-chain aminoacids (BCAA) are reported to be effective in prolongation of the mean survival time of patients with liver cirrhosis, it is not clear whether BCAA could influence the immune function in those patients. METHODS: Branched-chain amino acids were given as a supplement to carbon tetrachloride-induced cirrhotic rats, and an aminogram of the liver and kinetics of liver-associated lymphocytes (LAL) were then analysed. RESULTS: Liver cirrhosis was established at the 12th week, and glutathione S-transferase placental form (GST-P)-positive lesions, which are known to be pre-neoplastic lesions, occupied 1.72+/-0.84% of the liver at the 16th week in the controls. At this time the LAL showed an increase in the number of CD5-, CD8- and CD18 positive cells and augmentation of lectin-dependent cellular cytotoxicity (LDCC) activity. Furthermore, supplementation of BCAA increased the number of LAL, especially CD8-positive cells and natural killer cells, and augmented LDCC activity of LAL at the 16th week. The number of LAL was positively correlated with the valine concentration in the plasma and liver, and the area of GST-P positive lesions tended to be decreased in the BCAA group. CONCLUSION: These findings suggest that BCAA administration has stimulatory effects on the local immune systems of the liver, which may have a potential to inhibit hepatocarcinogenesis. Moreover, among all amino acids valine might be an important amino acid for enhancing the immune function of LAL. PMID- 11022825 TI - Outcomes of primary hepatocellular carcinoma treatment: an 8-year experience with 368 patients in Thailand. AB - BACKGROUND AND AIMS: Primary hepatocellular carcinoma (HCC) is common in Thailand and its prognosis is extremely poor. Hepatic resection and liver transplantation are modes of curative therapy and various therapies have been developed to treat inoperable HCC. This study was performed to determine the outcome of therapy and to identify prognostic factors for survival among Thai patients with HCC. METHODS: Three hundred and sixty-eight patients with tissue-proven or serodiagnosed HCC were studied. Differences in laboratory investigations, patient survival, treatment outcomes and prognostic factors were analysed. The Kaplan Meier method and the log-rank test were used to calculate and compare survival of patients receiving different therapies. RESULTS: Of the 368 patients 317 were males (86.1%, M: F ratio 6.2:1). The median age of all patients was 52 years range (2-85 years). Fifty-nine patients were specifically treated and 209 were conservatively treated. Among those tested, 70% had hepatitis B virus-related HCC. The overall median survival (range) after diagnosis and that of the treated and untreated patients were 5.6 (0-92.1), 9.0 (0-92.1) and 2.3 (0-25.9) months, respectively. More than 50% of the Okuda I patients survived beyond 30 months. The Okuda II and III patients had a median survival (range) of 5.7 (0.1-75.2) and 1.6 (0.1-25.9) months, respectively. Those patients treated by surgery, transcatheter oil chemoembolization and systemic chemotherapy had a better survival rate than did the conservatively managed patients. Surgery was found to be a better treatment than systemic chemotherapy. However, no differences between other pairs of treatment groups were identified. Subgroup analysis of the Okuda II group patients revealed the same results. Independent predictors of death were multiple lesions, advanced Okuda stage and treatment modality. CONCLUSION: Despite various kinds of therapy, the treatment outcomes for Thai HCC patients remain poor and largely depend on the disease extent and treatment received. Most Thai HCC patients are at an advanced stage at diagnosis. Thai patients who received disease-specific treatment other than intraarterial chemotherapy had a better survival rate than did supportive treatment patients. PMID- 11022826 TI - Bilirubin and bile acids may modulate their own metabolism via regulating uridine diphosphate-glucuronosyltransferase expression in the rat. AB - BACKGROUND AND AIMS: Uridine diphosphate (UDP)-glucuronosyltransferase (UGT) is a critical enzyme in the elimination of bilirubin and it also plays a role in the metabolism of bile acids. The aim of this study was to determine whether bilirubin and bile acids could modulate their own metabolism by regulating UGT levels in cultured rat hepatocytes. METHODS AND RESULTS: Incubation of hepatocytes with bilirubin (48 micromol/L) for 24 h significantly increased the mRNA expression of UGT1A1 and UGT1A5, two UGT isoforms responsible for the conjugation of bilirubin. The induction of UGT1A1 and UGT1A5 by bilirubin was concentration and time dependent. Treatment with chenodeoxycholic acid, cholic acid, deoxycholic acid, hyodeoxycholic acid and lithocholic acid at a concentration of 100 micromol/L for 48 h significantly enhanced the mRNA expression of UGT2B1, a UGT isoform responsible for the glucuronidation of bile acids. The UGT2B3 mRNA level was also increased by hyodeoxycholic acid. The regulation of UGT2B1 mRNA by chenodeoxycholic acid and hyodeoxycholic acid was dose and time dependent. CONCLUSION: Our results suggest that bilirubin and bile acids can induce UGT expression and as a result, these compounds may modulate their own metabolism. Such regulation could play a compensatory role in the pathological increased concentrations of these compounds in some hepatobiliary diseases. PMID- 11022827 TI - Effects of simvastatin on hepatic cholesterol metabolism, bile lithogenicity and bile acid hydrophobicity in patients with gallstones. AB - BACKGROUND AND AIMS: There is limited information available on the effects of 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitors on hepatic and biliary cholesterol metabolism in patients with gallstones. The aims of this study were to determine the effect of simvastatin on the regulatory elements of cholesterol metabolism that determine the concentrations of cholesterol in plasma and bile. METHODS: Thirty-one gallstone patients were enrolled in the study; 17 were treated with 20 mg simvastatin daily for 3 weeks prior to cholecystectomy and 14 served as controls. Samples of blood, liver, gall-bladder bile and bile from the common bile duct (CBD) were collected and analysed. RESULTS: The plasma cholesterol (-30%), triacylglycerol (-23%) and low-density lipoprotein (LDL) cholesterol (-42%) concentrations were significantly lowered by simvastatin treatment, as was the plasma lathosterol: cholesterol (-70%), which reflects whole-body cholesterol synthesis. Despite these changes, the hepatic LDL receptor protein and LDL receptor activity in circulating mononuclear cells were similar in both groups. There were no differences in the plasma phytosterol: cholesterol, which reflects the intestinal cholesterol absorption capacity or in the activity of hepatic acyl-coenzyme A: cholesterol acyltransferase. There were however, lower cholesterol concentrations in CBD (-68%) and gall bladder (-41%) bile, and decreased lithogenic (-47%) and bile acid hydrophobicity (-22%) indices of CBD bile in the simvastatin group. CONCLUSIONS: These data indicate that simvastatin reduced plasma and biliary cholesterol levels primarily by reducing cholesterol synthesis. The reduction in CBD bile lithogenicity and bile acid hydrophobicity by simvastatin suggests that this agent may be useful for people who have early stages of cholesterol gallstone development and in whom a choleretic effect is required. PMID- 11022828 TI - Role of neutrophils in sinusoidal endothelial cell injury after extensive hepatectomy in cholestatic rats. AB - BACKGROUND AND AIMS: The authors have shown previously that sinusoidal endothelial cell injury developed concomitantly with the accumulation of neutrophils in the hepatic sinusoidal space in cholestatic rats after extensive hepatectomy. The aim of the present study was to investigate the role of neutrophils in the development of this kind of endothelial cell injury. METHODS: Rats underwent 78% partial hepatectomy after 2 weeks of cholestasis, and subsequent external biliary drainage for 5 days. To decrease the number of neutrophils, antirat neutrophil serum was administered intraperitoneally. Some serum parameters and histological specimens were examined 48 h after partial hepatectomy. RESULTS: Anti-neutrophil serum significantly reduced the number of accumulated neutrophils in the hepatic sinusoids. Although the purine nucleoside phosphorylase: alanine aminotransferase ratio, a marker of non-parenchymal cell injury, was increased in cholestatic-hepatectomized rats, this abnormality was significantly attenuated by the treatment with antineutrophil serum. Electron microscopically, focal detachment of cytoplasms of sinusoidal endothelial cells was observed occasionally in cholestatic-hepatectomized rats, but was not found in the antirat neutrophil serum-treated rats. CONCLUSION: These results indicate that accumulated neutrophils might be important effector cells in the pathogenesis of sinusoidal endothelial cell injury after extensive hepatectomy in cholestatic rats, even after appropriate external biliary drainage. PMID- 11022829 TI - Biliary excretory function is regulated by canalicular membrane fluidity associated with phospholipid fatty acyl chains in the bilayer: implications for the pathophysiology of cholestasis. AB - BACKGROUND AND AIMS: Bile canalicular membrane fluidity is modulated by phospholipid molecular species within membrane lipid bilayers. Thus, organellar membrane lipid composition is a determinant of canalicular function. In this study, the effect of phalloidin-induced cholestasis on bile lipid composition and liver subcellular membrane fraction composition in rats was examined to clarify the relationship between cholestasis and hepatic lipid metabolism. METHODS AND RESULTS: Each rat received one phalloidin dose (400 microg/kg, i.v.). After the bile was collected, liver microsomes and canalicular membranes were analysed. The bile flow rate decreased by 50% 3.5 h after phalloidin administration. Although the bile acid output remained almost the same, the phospholipid and cholesterol output were significantly decreased (by 40.3+/-5.97% and 76.9+/-5.56%, respectively). Thus, the cholesterol:phospholipid (C:P) ratio in bile was significantly decreased by 80.4+/-10.1%. Phalloidin administration also increased the saturated: unsaturated fatty acid ratio (S:U) in bile for phosphatidylcholine by 25.5+/-3.2%. In the canalicular membrane, the C:P and S:U ratios for phosphatidylcholine were increased (24.8+/-4.2% and 34.4+/-6.9%, respectively), while the S:U for sphingomyelin was decreased by 61.0+/-6.2%. In microsomes, the C:P was decreased by 41.0+/-6.0%, but the S:U for both phosphatidylcholine and sphingomyelin were unaffected. Canalicular membrane fluidity, assayed by 1,6 diphenyl-1,3,5-hexatriene fluorescence depolarization, decreased significantly. Therefore, increased secretion of hydrophobic phosphatidylcholine into bile was associated with more hydrophobic canalicular membrane phosphatidylcholine, while sphingomyelin in the canalicular membrane was less hydrophobic. CONCLUSIONS: These results indicate that phalloidin uncouples secretion of cholesterol and phospholipids, which causes a redistribution of fatty acyl chain species among canalicular membrane phospholipids that alters membrane fluidity. These changes may be a homeostatic response mediated by the phospholipid translocator in the canalicular membrane, although direct evidence for this is unavailable. PMID- 11022830 TI - Changes in prostaglandin and nitric oxide levels in the hyperdynamic circulation following liver resection. AB - BACKGROUND AND AIMS: Significant changes in hepatic haemodynamics occur after major hepatectomy, but the pathogenesis of this phenomenon is unclear. This study investigates the serum profile of prostaglandin and nitric oxide in the hepatic and systemic circulation before and after hepatectomy and the temporal relationship of these to changes in the hepatic blood flow. METHODS: Blood samples were collected from the internal jugular, portal and hepatic veins of six Yorkshire pigs (17-25 kg) before, immediately after and 48 h after partial hepatectomy. RESULTS: Serum levels of prostacyclin I2 (PGI2) and prostaglandin E2 (PGE2) from the systemic circulation, the portal circulation and from the hepatic veins were found to differ considerably even before hepatectomy. After the hepatectomy was performed, there was a significant rise in PGI2 levels in the systemic circulation (P=0.027). Hepatic blood flow and cardiac output were measured before hepatectomy, 24 and 48 h after hepatectomy in another six pigs. A significant increase in hepatic blood flow (P=0.029) occurred after the hepatectomy and this was accompanied by a concomitant increase in the cardiac output (P=0.042). The increase in PGI2 concentration in the systemic circulation after hepatectomy appears to accompany the development of hyperdynamic hepatic and systemic circulations. No significant change was found in circulating PGE2 levels in the systemic, portal and hepatic veins at the three collection intervals (i.e. pre, post and 48 h after hepatectomy). The PGE2 levels in the hepatic vein, however, were significantly higher from than in the portal vein (P=0.028). No significant changes were observed in the level of nitric oxide. CONCLUSION: This study demonstrates an increase in the total hepatic blood flow after hepatectomy together with a threefold increase in prostacyclin in the systemic circulation. The liver was identified as the main source of circulatory prostaglandin. PMID- 11022831 TI - Oesophageal pH monitoring in infants: elimination of gastric buffering does not modify reflux index. AB - BACKGROUND: Oesophageal pH monitoring is considered to be the gold standard investigation in the diagnosis of gastro-oesophageal reflux disease. Results of pH monitoring in regurgitating infants, however, may be within normal ranges. Therefore it was hypothesized that prolonged buffering of gastric acidity caused by milk-feeding may falsely normalize the pH data. Therefore, it may be relevant to omit the periods of gastric acid buffering in the analysis of the oesophageal pH monitoring data. METHODS: Combined gastric and oesophageal pH monitoring (Digitrapper Mark III, two-channel antimony electrode, Synectics) was performed in 90 consecutive infants, 0.5-10 months old, who were referred to the unit by outdoor paediatricians for pH measurement. The data were analysed in three different ways. The reflux index (RI), or the percentage time that the pH in the oesophagus was < 4.0, was calculated in three different ways: (i) the total duration of the investigation; (ii) excluding 90-min postprandial periods, starting from the beginning of a feeding; and (iii) excluding all periods with a gastric pH > 4.0, which are the periods of gastric buffering with a theoretical impossibility of recording an oesophageal pH < 4.0. RESULTS: The mean duration of the pHmetries was 19.20+/-2.01h (A; mean +/- 1 SD), with a RI of 5.38+/-5.39% (B; median 3.45%). If the 90-min postprandial periods were not considered, the mean duration decreased to 12.21+/-2.41h (C; P(A vs C) < 0.001), with an oesophageal RI of 6.82+/-6.57% (D; median: 4.65; P(B vs D) = NS; r (B vs D) = 0.97). If all periods with a gastric pH > 4.0 were elminated, the duration available for analysis of the data with a gastric pH < 4.0 was 12.53+/-4.00 h (E; P(A vs E) < 0.001; P(C vs E) = NS), with an oesophageal RI of 7.06+/-7.52% (F; median: 4.50; P(B vs D vs F) = NS; r(B vs F), (D vs F) = 0.96). CONCLUSION: Overall the data do not support a benefit from including periods of gastric buffering in the routine analysis of oesophageal pH monitoring data. Therefore standard oesophageal pH monitoring should be with a single electrode and should not include periods of gastric buffering. PMID- 11022833 TI - Efficacy of a 1-week pantoprazole triple therapy in eradicating Helicobacter pylori in Asian patients. AB - BACKGROUND: The aim of the present paper was to determine the efficacy and tolerability of a 1-week treatment regimen consisting of pantoprazole and two antibiotics: clarithromycin and amoxycillin, in the eradication of Helicobacter pylori. METHODS: The patients selected had unequivocal evidence of H. pylori infection based on urease test, culture and histology on antral and corpus biopsies obtained at endoscopy. Patients received pantoprazole 40 mg twice a day, clarithromycin 500 mg twice a day and amoxycillin 1 g twice a day for 1 week and were assessed for successful eradication at least 4 weeks after completion of therapy by repeat gastroscopy and gastric biopsies. Eradication was defined as absence of bacteria in both antral and corpus biopsies tested by culture, histology and urease test. RESULTS: One hundred and six patients were recruited for the study. The mean age was 48.0 years (range: 23-74 years). Four patients defaulted follow up and five patients were not compliant (taking less than 85%) with medications. Eradication rates on per-protocol analysis were: 88/97 (90.7%; 95% CI: 83.1-95.7); and on intention-to-treat analysis they were: 88/106 (83.0%; 95% CI: 75.9-90.2). Side-effects were in general mild and tolerable: 57 of 106 (53.7%) patients complained of a bitter taste; 15 (14.1%) complained of giddiness; 10 (9.4%) complained of increased abdominal pain; 11 (11.5%) complained of lethargy and 16 (15.1%) complained of loose motions. Pre-treatment metronidazole resistance was encountered in 57/74 strains (77.0%). Clarithromycin resistance was not encountered in any of the strains. CONCLUSIONS: The pantoprazole 1-week triple therapy with amoxycillin and clarithromycin is effective in H. pylori eradication. The treatment was well tolerated by patients. Metronidazole resistance was reported in a high percentage of strains isolated from patients. Clarithromycin resistance was, however, not detected in any of the strains. PMID- 11022832 TI - Clarithromycin-resistance and point mutations in the 23S rRNA gene in Helicobacter pylori isolates from Japan. AB - BACKGROUND: Resistance of Helicobacter pylori to clarithromycin is mostly due to the point mutations in the 23S rRNA. In Japan, however, the frequency of these mutations has not been fully investigated. Furthermore, no study has used gastric biopsy specimens to detect these point mutations. METHODS: The frequency of primary clarithromycin-resistant H. pylori was examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Eighty-two strains (42 isolated from patients with gastric cancer and 40 isolated from patients with chronic gastritis) were examined. Two biopsy specimens obtained from patients in whom eradication therapy including clarithromycin had failed were also studied. RESULTS: Either A2143G or A2144G point mutation was detected in 90% of clarithromycin-resistant H. pylori strains. Eight out of 82 strains (9.8%) had either A2143G or A2144G point mutation. Only one out of 42 strains in patients with gastric cancer had A2143G mutation, whereas five strains had A2144G and two had A2143G mutations in 40 strains isolated from control subjects. The proportion was significantly lower in patients with early gastric cancer (P < 0.05). This PCR-RFLP was also applicable for DNA samples extracted from biopsy specimens and infection of clarithromycin-resistant H. pylori was observed. CONCLUSION: The results suggest that the point mutation in the 23S rRNA gene is commonly seen in clarithromycin-resistant H. pylori and it contributes to the treatment failure in Japan. The PCR-RFLP system is a sensitive method by which to diagnose H. pylori infection as well as a simple method for detecting clarithromycin resistance without bacterial culture. PMID- 11022834 TI - L-Arginine protects and exacerbates ethanol-induced rat gastric mucosal injury. AB - BACKGROUND AND AIMS: We have previously demonstrated that 60% ethanol (EtOH) increases macromolecular leakage and induces focal lesion formation in areas of permanent flow stasis within gastric mucosal vessels. Nitric oxide (NO) may prevent lesion formation by inhibiting leakage. This study used fluorescent in vivo microscopy to investigate (i) whether L-arginine (NO precursor) prevented EtOH induced injury and (ii) the mechanisms of protection. METHODS: Experiments were carried out on anaesthetized rats (hypnorm/diazepam) receiving either intra arterial fluorescein isothiocyanate-bovine serum albumin (0.2 mL/100 g), a marker for quantitating leakage, or Acridine red (0.1 mL/100 g) which labels leukocytes. Animals then received 100, 300 or 500 mg/kg L-arginine (i.a.) followed by 60% EtOH or distilled water, topically applied to the gastric mucosa (n = 6 for each group). Vessel diameter, macromolecular leakage of labelled albumin from post capillary venules (PCV) and capillaries and leukocyte activity were quantitated using image analysis. RESULTS: L-Arginine (100 mg/kg) did not increase vessel diameter or prevent EtOH-induced lesion formation and leakage. Both 300 mg/kg and 500 mg/kg alone induced significant and sustained increases in PCV diameter after 15 (P< 0.01) and 5 min (P< 0.001), respectively. Lesion formation was only prevented by 300 mg/kg L-arginine, whereas 500 mg/kg exacerbated haemorrhagic lesion formation over the entire exposed mucosa. Neither 300 mg/kg nor 500 mg/kg L-arginine prevented leakage following EtOH. No leukocyte activity was observed following EtOH with or without L-arginine pretreatment. CONCLUSION: L-Arginine (300 mg/kg) prevented lesion formation. The mechanism of protection probably involved the increased blood flow in the dilated PCV and not the inhibition of leakage. The combined effects of EtOH and the possible high NO levels exacerbate gastric mucosal damage despite the increases observed in vessel diameter. PMID- 11022835 TI - Effect of a calcium channel blocker and antispasmodic in diarrhoea-predominant irritable bowel syndrome. AB - BACKGROUND: Irritable bowel syndrome (IBS) is a colonic function disorder. Both pinaverlum bromide (a selective calcium channel blocker) and mebeverine (an antispasmodic) are reported to be effective in the long-term (12-16 weeks) treatment of IBS patients. Their efficacy in the short-term treatment of IBS patients and colonic transit time is unclear. Furthermore, substance P and neuropeptide Y have either excitatory or inhibitory effects on colonic motility. Whether the efficacy of both drugs is mediated through these neuropeptides remains unknown. METHODS AND RESULTS: A clinical trial was conducted with 91 patients with diarrhoea-predominant IBS. After basal measurement of the total colonic transit time, IBS patients were randomized to receive either pinaverlum bromide (50 mg, t.i.d.) or mebeverine (100 mg, t.i.d.) for 2 weeks. The symptomatic scores regarding defaecation, total colonic transit time and serum levels of substance P and neuropeptide Y were measured before and after treatments. The daily defaecation frequency was markedly decreased after treatment (pinaverlum bromide, 2.9+/-1.2 vs 2.0+/-1.0, P< 0.05; mebeverine, 2.7+/ 1.1 vs 2.1+/-1.0, P< 0.05). The stool consistency became well formed after both treatments (P< 0.05). Both drugs similarly improved the global well-being in these IBS patients (pinaverlum bromide vs mebeverine 73.4 vs 71.8%, P> 0.05). The total colonic transit time was significantly prolonged only after pinaverlum bromide treatment (21.4+/-15.5 vs 30.8+/-14.8 h, P< 0.01). Neither substance P nor neuropeptide Y serum level was significantly changed after either treatments. CONCLUSION: Pinaverlum bromide and mebeverine have similar therapeutic efficacies on diarrhoea-predominant IBS patients. Prolonged colonic transit time may be one of the factors responsible for the efficacy of pinaverlum bromide on the IBS patients. Substance P and neuropeptideY appear less important in the pathogenesis of diarrhoea-predominant IBS. PMID- 11022836 TI - Effect of argatroban on trinitrobenzene sulfonic acid-induced colitis. AB - BACKGROUND: Recent studies have suggested that heparin is effective for treatment of inflammatory bowel disease (IBD) and its various effects (in addition to the anticoagulant effect). We evaluated the effects of argatroban as an antithrombin drug on trinitrobenzene sulfonic acid (TNB)-induced colitis, an established model of IBD. METHODS: Rats were randomly assigned to four groups in which mini-osmotic pumps containing saline (TNB-S), argatroban (TNB-A), or 100 U/kg heparin (TNB-H) were intraperitoneally implanted. Three days after the pumps were implanted, TNB was infused via the anus, and colitis was induced. After 5 days, prothrombin time (PT), activated partial prothrombin time (APTT), antithrombin III (AT-III), platelet, fibrinogen, colonic wet weight, macroscopic damage score, histological score, mucosal myeloperoxidase activity and mucosal leukotrien B4 (LTB4) levels were compared among the four groups. RESULTS: The APTT was prolonged in the heparin treatment group but only slightly prolonged in the argatroban treatment group. The platelet count and the fibrinogen level were higher in the TNB-S group than in the healthy control group and the AT-III level was slightly lower in the TNB-S group than in the healthy control group and lower still in the TNB-H group. CONCLUSIONS: The colonic wet weight was similar among the four groups while the macroscopic damage score, histological score, mucosal myeloperoxidase activity and the mucosal LTB4 level were significantly decreased in the TNB-A and TNB-H groups. Argatroban, as well as heparin may be effective for treatment of TNB induced colitis. PMID- 11022837 TI - Yield from colonoscopic screening in people with a strong family history of common colorectal cancer. AB - BACKGROUND AND AIMS: People with a strong family history of common (so-called 'sporadic') colorectal cancer are generally advised to undergo colonoscopic screening, but the starting age for this is unclear. An audit was performed to study the age-related yield of screening colonoscopy in this risk group. METHODS: A prospective audit of the outcome of screening colonoscopy was performed on a cohort of 232 people with a strong family history of common colorectal cancer. All were registrants in a familial bowel cancer service solely because of their family medical history. They had no bowel symptoms and no prior endoscopic investigation of the large bowel. RESULTS: Neoplastic lesions were detected by using colonoscopy in 33 participants. In 27 participants, the major lesion was a small tubular adenoma, four had an advanced adenoma and two had cancer. More neoplastic (P= 0.02) and advanced neoplastic (P= 0.03) lesions were found in those patients aged > or = 50 years. Only one advanced adenoma was detected in a participant below the age of 50 years. CONCLUSION: The yield from screening colonoscopy in young people (< 50 years) with a strong family history of common colorectal cancer is low, placing doubt on the need for colonoscopic screening before the age of 50 years. PMID- 11022839 TI - Hepatitis G virus genotypes in Australia, Papua New Guinea and the Solomon Islands: a possible new Pacific type identified. AB - BACKGROUND: Hepatitis G Virus (HGV)/GB Virus-C (GBV-C) is a newly discovered RNA virus. Nucleotide sequence comparison and phylogenetic studies of the 5' untranslated region (5'UTR) within the viral genome have identified at least three different types which have provisionally been classified as type 1 (West African origin), type 2 (North American origin) and type 3 (Asian origin). METHODS AND RESULTS: The products of RT-PCR were sequenced by using blood donors and patients infected with HGV/GBV-C in Australia, Papua New Guinea and the Solomon Islands to investigate the genotype distribution in this area of the world. All the Australian isolates showed strong sequence homology with type 2, while the Papua New Guinea and Solomon Islands sequences were more closely related, but differ from type 3, which has previously been reported from isolates studied within Asia. CONCLUSIONS: Phylogenetic analysis suggests that these latter sequences are either a new HGV/GBV-C Pacific type or a subtype of the Asian type RNA virus. Isolates homologous with type 1 were not identified in these population groups. PMID- 11022838 TI - Measurement of serum hyaluronic acid in patients with chronic hepatitis C and its relationship to liver histology. Consensus Interferon Study Group. AB - BACKGROUND AND AIMS: Chronic hepatitis C is a slowly progressing inflammatory disease of the liver that can lead to cirrhosis and its complications. Assessment of liver damage in hepatitis C has been primarily via histological evaluation. Liver biopsy, while useful in determining the extent of liver damage, has associated costs and places patients at a small but finite risk of bleeding. Studies in small patient populations have identified serum markers shown to correlate with liver histology, including procollogen III peptide and hyaluronic acid (HA). To determine whether serum HA was a reliable predictor of cirrhosis and fibrosis, we examined serum HA concentrations from 486 chronic Hepatitis C virus (HCV) patients. METHODS AND RESULTS: Patients were anti-HCV and HCV RNA positive, with elevated alanine aminotransferase values and underwent a liver biopsy. Sera were obtained at the baseline for HA using radioimmunoassay methodology. Patients with cirrhosis had significantly higher serum HA concentrations compared with non-cirrhotic patients (382+/-31 vs 110+/-9 microg/L respectively, P< 0.001). Patients with fibrosis had significantly higher mean serum HA concentrations (179+/-11 microg/L) compared with patients without fibrosis (62+/-20 microg/L; P< 0.001). The correlation between HA concentration and the components of the Knodell histological activity index score revealed no strong associations with the exception of fibrosis, which showed moderate correlation (R=0.5421, P<0.001). The clinical value of HA measurement appears to be its ability to exclude cirrhosis. A HA value of < 60 microg/L excluded the presence of cirrhosis or significant fibrosis with a predictive value of 99 and 93%, respectively. CONCLUSIONS: Serum HA measurement may be clinically useful to non-invasively assess the degree of fibrosis and cirrhosis. Further prospective studies are warranted to determine the clinical utility of HA as a non-invasive marker of liver fibrosis. PMID- 11022840 TI - Hepatobiliary and pancreatic: a boy with ascites. PMID- 11022841 TI - Gastrointestinal: Crohn's disease of the oesophagus. PMID- 11022842 TI - Transjugular intrahepatic portosystemic shunt for intractable colonic bleeding in a patient with cirrhosis. AB - A patient with portal hypertension due to alcoholic liver disease developed colonic bleeding that responded to transjugular intrahepatic portosystemic shunt (TIPS) after the failure of other measures. This case illustrates that TIPS should be considered in the treatment of colonic, as well as upper gastrointestinal, bleeding associated with portal hypertension. PMID- 11022843 TI - Hemoglobin-based blood substitutes and the hazards of blood radicals. AB - Cell-free hemoglobins, chemically altered or genetically expressed in microbial host systems, have been developed as oxygen-carrying therapeutics. Site-directed modifications are introduced and serve to stabilize the protein molecules in a tetrameric and/or a polymeric functional form. Animal studies, as well as recent clinical studies, have suggested these proteins probably deliver oxygen to tissues. However, concerns still persist regarding the interference of hemoglobin and its oxidation products with the vascular redox balance, potentially impeding its clinical usefulness. This article reviews our current understanding of heme mediated toxicities and some of the emerging protective strategies used to overcome hemoglobin side reactions. PMID- 11022844 TI - 4-Hydroxynonenal, an end-product of lipid peroxidation, induces apoptosis in human leukemic T- and B-cell lines. AB - 4-Hydroxynonenal (HNE) is the major aldehydic product resulting from lipid peroxidation and has been implicated as involved in several pathological conditions. In our continuing studies on the role of membranes and lipid peroxidation in the induction of apoptosis, we investigated the effect of HNE on cultured human malignant immune system cells. Two cell lines were utilized; MOLT 4, a human T-cell leukemia cell line, and Reh, a human B-cell lymphoma cell line. A 10 min treatment with 0.01 mM HNE resulted in the apoptotic death, as determined by flow cytometric and morphological analyses, of both cell lines within 24 h. MOLT-4 cells exhibited the manifestations of impending apoptotic death much sooner than did Reh cells, indicating that MOLT-4 cells were more sensitive or not as efficient at detoxifying HNE than were Reh cells. These results suggest that peroxidative damage to cellular membranes resulting in the production of HNE may be a trigger for the induction of apoptosis in immune system cells. PMID- 11022845 TI - Cu2+-catalyzed oxidative degradation of thyroglobulin. AB - Thyroglobulin (Tg) was subjected to metal-catalyzed oxidation, and the oxidative degradation was analyzed by SDS-polyacrylamide gel electrophoresis under reducing conditions. In contrast to no effect of hydrogen peroxide (H2O2) alone on the Tg degradation, the inclusion of Cu2+ (30 microM), in combination with 2 mM H2O2, caused a remarkable degradation of Tg, time- and concentration-dependent. The action of Cu2+ was not mimicked by Fe2+, suggesting that Tg may interact selectively with Cu2+. A similar degradation of Tg was also observed with Cu2+/ascorbate system, and the concentration of Cu2+ (5-10 microM), in combination with ascorbate, required for the effective degradation was smaller than that of Cu2+ (10-30 microM) in combination with H2O2. In support of involvement of H2O2 in the Cu2+/ascorbate action, catalase expressed a complete protection. However, hydroxyl radical scavengers such as dimethylsulfoxide or mannitol failed to prevent the oxidation of Tg whereas phenolic compounds, which can interact with Cu2+, diminished the oxidative degradation, presumably consistent with the mechanism for Cu2+-catalyzed oxidation of protein. Moreover, the amount of carbonyl groups in Tg was increased as the concentration (3-100 microM) of Cu2+ was enhanced, while the formation of acid-soluble peptides was not remarkable in the presence of Cu2+ up to 200 microM. In further studies, Tg pretreated with heat or trichloroacetic acid seemed to be somewhat resistant to Cu2+-catalyzed oxidation, implying a possible involvement of protein conformation in the susceptibility to the oxidation. Based on these observations, it is proposed that Tg could be degraded non-enzymatically by Cu2+-catalyzed oxidation. PMID- 11022846 TI - Immunochemical quantitation of UV-induced oxidative and dimeric DNA damage to human keratinocytes. AB - There is growing evidence to suggest that solar radiation-induced, oxidative DNA damage may play an important role in skin carcinogenesis. Numerous methods have been developed to sensitively quantitate 8-oxo-2'deoxyguanosine (8-oxodG), a recognised biomarker of oxidative DNA damage. Immunoassays may represent a means by which the limitations of many techniques, principally derived from DNA extraction and sample workup, may be overcome. We report the evaluation of probes to thymine dimers and oxidative damage in UV-irradiated cells and the DNA derived therefrom. Thymine dimers were most readily recognised, irrespective of whether in situ in cells or in extracted DNA. However, using antibody-based detection the more subtle oxidative modifications required extraction and, in the case of 8 oxodG, denaturation of the DNA prior to successful recognition. In contrast, a recently described novel probe for 8-oxodG detection showed strong recognition in cells, although appearing unsuitable for use with extracted DNA. The probes were subsequently applied to examine the relative induction of lesions in cells following UV irradiation. Guanine-glyoxal lesions predominated over thymine dimers subsequent to UVB irradiation, whereas whilst oxidative lesions increased significantly following UVA irradiation, no induction of thymine dimers was seen. These data support the emerging importance of oxidative DNA damage in UV-induced carcinogenesis. PMID- 11022847 TI - Interactions of superoxide anion with enzyme radicals: kinetics of reaction with lysozyme tryptophan radicals and corresponding effects on tyrosine electron transfer. AB - The kinetics of O2*- reaction with semi-oxidized tryptophan radicals in lysozyme, Trp*(Lyz) have been investigated at various pHs and conformational states by pulse radiolysis. The Trp*(Lyz) radicals were formed by Br2*- oxidation of the 3 4 exposed Trp residues in the protein. At pH lower than 6.2, the apparent bimolecular rate is about 2 x 10(8) M(-1) s(-1); but drops to 8 x 10(7) M(-1) s( 1) or less above pH 6.3 and in CTAC micelles. Similarly, the apparent bimolecular rate constant for the intermolecular Trp*(Lyz) + Trp*(Lyz) recombination reaction is about (4-7 x 10(6) M(-1) s(-1)) at/or below pH 6.2 then drops to 1.3-1.6 x 10(6) M(-1) s(-1) at higher pH or in micelles. This behavior suggests important conformational and/or microenvironmental rearrangement with pH, leading to less accessible semi-oxidized Trp* residues upon Br2*- reaction. The kinetics of Trp*(Lyz) with ascorbate, a reducing species rather larger than O2*- have been measured for comparison. The well-established long range intramolecular electron transfer from Tyr residues to Trp radicals--leading to the repair of the semi oxidized Trp*(Lyz) and formation of the tyrosyl phenoxyl radical is inhibited by the Trp*(Lyz) + O2*- reaction, as is most of the Trp*(Lyz) + Trp*(Lyz) reaction. However, the kinetic behavior of Trp*(Lyz) suggests that not all oxidized Trp residues are involved in the intermolecular recombination or reaction with O2*-. As the kinetics are found to be quite pH sensitive, this study demonstrates the effect of the protein conformation on O2*- reactivity. To our knowledge, this is the first report on the kinetics of a protein-O2*- reaction not involving the detection of change in the redox state of a prosthetic group to probe the reactivity of the superoxide anion. PMID- 11022848 TI - Role of reactive oxygen species in cardiac preconditioning: study with photoactivated Rose Bengal in isolated rat hearts. AB - Oxygen radical scavengers have been shown to prevent the development of ischemic preconditioning, suggesting that reactive oxygen species (ROS) might be involved in this phenomenon. In the present study, we have investigated whether direct exposure to ROS produced by photoactivated Rose Bengal (RB) could mimic the protective effects of ischemic preconditioning. METHODS: In vitro generation of ROS from photoactivated RB in a physiological buffer was first characterised by ESR spectroscopy in the presence of 2,2,6,6-tetramethyl-1-piperidone (oxoTEMP) or 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). In a second part of the study, isolated rat hearts were exposed for 2.5 min to photoactivated RB. After 5 min washout, hearts underwent 30 min no-flow normothermic ischemia followed by 30 min of reperfusion. RESULTS AND CONCLUSIONS: The production of singlet oxygen (1O2) by photoactivated RB in the perfusion medium was evidenced by the ESR detection of the nitroxyl radical oxoTEMPO. Histidine completely inhibited oxoTEMPO formation. In addition, the use of DMPO has indicated that (i) superoxide anions (O2*-) are produced directly and (ii) hydroxyl radicals (HO*) are formed indirectly from the successive O2*- dismutation and the Fenton reaction. In the perfusion experiments, myocardial post-ischemic recovery was dramatically impaired in hearts previously exposed to the ROS produced by RB photoactivation (1O2, O2*-, H2O2 and HO*) as well as when 1O2 was removed by histidine (50 mM) addition. However, functional recovery was significantly improved when hearts were exposed to photoactivated RB in presence of superoxide dismutase (10(5) IU/L) and catalase (10(6) IU/L). Further studies are now required to determine whether the cardioprotective effects of Rose Bengal in presence of O2*- and H2O2 scavengers are due to singlet oxygen or to other species produced by Rose Bengal degradation. PMID- 11022849 TI - Mechanism of alpha-tocopheryl succinate-induced apoptosis of promyelocytic leukemia cells. AB - Selective induction of apoptosis in tumor cells is important for treating patients with cancer. Because oxidative stress plays an important role in the process of apoptosis, we studied the effect of alpha-tocopheryl succinate (VES) on the fate of cultured human promyelocytic leukemia cells (HL-60). The presence of fairly low concentrations of VES inhibited the growth and DNA synthesis of HL 60 cells, and also induced their apoptosis via a mechanism that was inhibited by z-VAD-fluoromethylketone (z-VAD-fmk), an inhibitor of pan-caspases. VES activated various types of caspases, including caspase-3, 6, 8, and 9, but not caspase-1. VES triggered the reaction leading to the cleavage of Bid, a member of the death agonist Bcl-2 family, and released cytochrome c (Cyt.c) from the mitochondria into the cytosol by a z-VAD-fmk-inhibitable mechanism. VES transiently increased the intracellular calcium level [Ca2+]i and stimulated the release of Cyt.c in the presence of inorganic phosphate (Pi). However, high concentrations of VES (approximately 100 microM) hardly induced swelling of isolated mitochondria but depolarized the mitochondrial membrane potential by a cyclosporin A (CsA) insensitive mechanism. These results indicate that VES-induced apoptosis of HL-60 cells might be caused by activation of the caspase cascade coupled with modulation of mitochondrial membrane function. PMID- 11022850 TI - Flavonoids protect against oxidative damage to LDL in vitro: use in selection of a flavonoid rich diet and relevance to LDL oxidation resistance ex vivo? AB - The ability of a range of dietary flavonoids to inhibit low-density lipoprotein (LDL) oxidation in vitro was tested using a number of different methods to assess oxidative damage to LDL. Overall quercetin was the most effective inhibitor of oxidative damage to LDL in vitro. On this basis, a diet enriched with onions and black tea was selected for a dietary intervention study that compared the effect on the Cu2+ ion-stimulated lag-time of LDL oxidation ex vivo in healthy human subjects of a high flavonoid diet compared with a low flavonoid diet. No significant difference was found in the Cu2+ ion-stimulated lag-time of LDL oxidation ex vivo between the high flavonoid and low flavonoid dietary treatments (48 +/- 1.6 min compared to 49 +/- 2.1 min). PMID- 11022851 TI - Carotenoid oxidative degradation products inhibit Na+-K+-ATPase. AB - This study investigates the biological significance of carotenoid oxidation products using inhibition of Na+-K+-ATPase activity as an index. Beta-carotene was completely oxidized by hypochlorous acid and the oxidation products were analyzed by capillary gas-liquid chromatography and high performance liquid chromatography. The Na+-K+-ATPase activity was assayed in the presence of these oxidized carotenoids and was rapidly and potently inhibited. This was demonstrated for a mixture of beta-carotene oxidative breakdown products, beta Apo-10'-carotenal and retinal. Most of the beta-carotene oxidation products were identified as aldehydic. The concentration of the oxidized carotenoid mixture that inhibited Na+-K+-ATPase activity by 50% (IC50) was equivalent to 10 microM non-degraded beta-carotene, whereas the IC50 for 4-hydroxy-2-nonenal, a major lipid peroxidation product, was 120 microM. Carotenoid oxidation products are more potent inhibitors of Na+-K+-ATPase than 4-hydroxy-2-nonenal. Enzyme activity was only partially restored with hydroxylamine and/or beta-mercaptoethanol. Thus, in vitro binding of carotenoid oxidation products results in strong enzyme inhibition. These data indicate the potential toxicity of oxidative carotenoid metabolites and their activity on key enzyme regulators and signal modulators. PMID- 11022852 TI - Inter- and intra-individual variation in plasma and red blood cell vitamin E after supplementation. AB - To establish the range of individual blood responses to supplemental vitamin E, 30 healthy subjects ingested 75 mg of deuterium-labelled alpha-tocopherol with a standard breakfast. Blood was collected at 6, 9, 12, 27 and 51 h post ingestion and deuterated (d6) and non-deuterated (do) alpha-tocopherol concentrations were determined in plasma and red blood cells (RBC) by GC-MS. To examine intra individual responses, 6 of these subjects were re-examined at 6-month intervals over a 30-month period. Post ingestion, the amount of d6-alpha-tocopherol in blood increased rapidly with time with maximal concentrations seen at 12 h (plasma) and 27 h (RBC) in most subjects. At these times, d6-alpha-tocopherol concentration ranged from 0.3-12.4 micromol/l in plasma and 0.6-4.09 micromol/l packed cell in RBC. Area under the curve calculations indicated inter-individual differences of alpha-tocopherol uptake to be 40-fold for plasma (12.9-493.3 micromol h/l) and 6-fold for RBC (24.4-146.1 micromol h/l packed RBC). Intra individual variation in alpha-tocopherol uptake was small in comparison and remained relatively constant over the 30-month period. We conclude that vitamin E uptake varies widely in the normal population, although it is comparatively stable for an individual over time. These differences likely arise from variations in the regulation of vitamin E uptake and metabolism between subjects. Factors regulating this process must be better understood before the optimal intake of vitamin E can be ascertained. PMID- 11022853 TI - A meta-analysis of Parkinson's disease and exposure to pesticides. AB - This study examined the association between Parkinson's disease (PD) and exposure to pesticides. A series of meta-analysis of peer-reviewed studies were performed, using 19 studies published between 1989 and 1999. Prior to the meta-analysis, all studies were reviewed and evaluated for heterogeneity and publication bias. Significant heterogeneity among studies was detected and combined odds ratio (OR) was calculated using the random effect model. The majority of the studies reported consistent elevation in the risk of PD with exposure to pesticides. The combined OR studies was 1.94 [95% confidence interval (95% CI) 1.49-2.53] for all the studies, and 2.15 (95% CI 1.14-4.05) for studies performed in United States. Although the risk of PD increased with increased duration of exposure to pesticides, no significant dose-response relation was established, and no specific type of pesticide was identified. Our findings suggest that exposure to pesticides may be a significant risk factor for developing PD. PMID- 11022854 TI - Mitochondrial ND1 sequence analysis and association of the T4216C mutation with Parkinson's disease. AB - Mitochondrial dysfunction originating from mutations in Complex I genes may play a role in the pathogenesis of Parkinson's disease (PD). In this study, the entire ND1 coding sequence was sequenced in 84 newly diagnosed PD cases and 127 age/gender-matched controls. Numerous missense mutations were found at low frequency (<5%), whereas a thymidine to cytosine missense mutation at position 4216 that results in the replacement of tyrosine with histidine was found in 25% of the PD case samples and in 18% of the controls. When calculated according to gender, the 4216 mutation was observed in 26% of the male cases versus 16% of male controls (Odds Ratio [OR] = 1.85; 95% CI = 0.79-4.34). In contrast, females exhibited approximately equal frequencies among cases (22.5%) and controls (21%), yielding an OR of 1.08 (95% C.I. = 0.36-3.22). The findings indicate only a weak association of this genetic variant with PD. PMID- 11022855 TI - Neurofilaments are non-essential elements of toxicant-induced reductions in fast axonal transport: pulse labeling in CNS neurons. AB - Acrylamide (ACR) and g-diketones (g-DK) produce distal sensory-motor neuropathy in a variety of species, including humans. The specific molecular site and mechanism of toxicant action leading to specific morphological and behavioral abnormalities requires definition. The relative roles of fast anterograde axonal transport and neurofilaments (NF) are investigated using optic nerves of mice, with and without axonal neurofilaments. Segmental analysis, following pulse labeling with 3H-leucine into the vitreous body, was used to detect changes in fast anterograde transport in the optic nerve and tract. Single injections of ACR significantly reduced the quantity of radiolabeled proteins transported in both transgenic (lacking NF) and non-transgenic (containing NF) mice by 68.4% and 46.2%, respectively. Similarly, single injections of 2,5-hexanedione (2,5-HD) reduced the quantity of radiolabeled transport in transgenic and non-transgenic mice by 55.2% and 47.1%, respectively. Equimolar doses of propionamide and 3,4 hexanedione (non-neurotoxic analogues of ACR and 2,5-HD, respectively) produced no changes in the quantity or apparent rate of optic nerve transport. Additionally, no differences in quantity or apparent rate of transport between transgenic and non-transgenic animals were observed under control or experimental conditions. Therefore, ACR and 2,5-HD reduce the quantity of fast anterograde axonal transport in mouse CNS axons in a comparable amount to previously reported reductions in rat PNS axons. The absence of axonal neurofilaments had no effect on normal fast transport. Furthermore, the presence or absence of neurofilaments did not alter the effect of these toxicants on fast axonal transport. We conclude that toxicant-induced reductions in fast axonal transport are unrelated to ACR and g-diketone effects on NF or their accumulation. PMID- 11022856 TI - Residual neurologic deficits 30 years after occupational exposure to elemental mercury. AB - A battery of tests of peripheral and central nervous system function was administered to 205 former workers of a large heavy industrial plant, 104 of whom were previously exposed to inorganic mercury. The mean age of those examined was 71 years. Exposed subjects had participated in a urine-mercury exposure monitoring program during the time of operation of a process that required the use of mercury and its subsequent clean-up. Mercury exposure had been high (mean peak urine mercury concentration was >600 microg/l) and had ended 30 years or more prior to the investigation. Peripheral nerve function outcomes that were statistically significantly associated with cumulative mercury exposure after controlling for covariates included classification as having peripheral neuropathy, peroneal motor nerve conduction velocity, ulnar motor nerve conduction velocity, and peroneal motor nerve F-wave latency. Quantitative assessment of resting tremor was nearly significantly associated with cumulative mercury exposure (p=0.07). Among tests of central nervous system function, results of the Handeye Coordination test were significantly associated with cumulative mercury exposure after controlling for covariates. Cumulative mercury exposure was not observed to be associated with a quantitative measure of dementia or with a number of cognitive neurobehavioral test outcomes. The statistically significant associations with mercury exposure were observed in spite of greater mortality among the exposed group than the unexposed group. These results suggest that substantial occupational mercury exposure can have long-term adverse effects on the peripheral nervous system detectable decades after cessation of exposure. Such long-term adverse effects were not observed for a measure of dementia or other measures of cognitive function. PMID- 11022857 TI - Neurological effects of environmental exposure to arsenic in dust and soil among humans. AB - Persons living in a small town in Georgia, USA, were studied because of known environmental exposure to arsenic-containing dust. To assess the effects of this exposure on the nervous system, a cross-sectional epidemiological study was performed. The health outcome assessments of the study were based on a priori selected clinical examination results and quantitative electrophysiologic and behavioral outcome measures. Because the primary goal was to identify cases of peripheral neuropathy, a clinically relevant case definition, based on results of these outcomes was created. Historical exposure reconstructions were performed on a subset of the exposed subjects for whom such information was available. Of the 238 persons enrolled in the study, 133 were classified as non-exposed (i.e., did not reside in the area of known exposure) and 105 were classified as exposed (i.e., did reside in the area of known exposure). Following exclusions of persons with occupational exposure to neurotoxicants, pre-existing conditions associated with peripheral nerve impairment, or age below 18 years, 118 unexposed subjects and 85 exposed subjects were available for inclusion in the analyses of peripheral nerve outcomes. A total of 4 (3.4%) of the unexposed subjects and 13 (15.3%) of the exposed subjects met the case definitions for peripheral neuropathy (OR=5.1; p=0.004). Comparison of three exposure groups (none, low, high) did not yield statistically significant differences. Statistically significant exposure group differences were observed on quantitative tests of standing steadiness, vibrotactile threshold and tremor intensity but not for nerve conduction measures. These results demonstrate a strong association between community arsenic containing dust exposure and peripheral neuropathy among participants in this investigation. PMID- 11022858 TI - Visual and neurobehavioral impairment associated with polychlorinated biphenyls. AB - To determine whether residents exposed to PCBs and thermolysis products had impaired neurobehavioral functions, the performance of 98 exposed adults was compared with 58 unexposed regional referents (all volunteers). Visual field performance, color confusion index, balance as sway speed, blink reflex latency R 1, hearing, grip strength, simple and choice visual reaction times problem solving for Culture Fair and digit symbol, recall memory, peg placement, trail making A and B for attention and dexterity and long-term memory were tested. A profile of mood states (POMS) and questionnaires for chemical exposures, medical histories and the frequency of 35 symptoms were completed. Only statistically significant differences are described. Exposed subjects had slower simple and choice reaction times and faster sway speeds with eyes closed and open. Color discrimination and visual performance scores were lower and visual fields were often constricted. Scores on Culture Fair, digit symbol, vocabulary and verbal recall were lower. Placement of pegs in a slotted pegboard was slower and trail making A and B took longer. Even embedded memory test scores including vocabulary were lower. POMS scores were elevated. There were no competing chemical exposures, confounding factors or other explanations for the findings. Long residential exposure to PCBs was associated with visual defects and impaired neurophysiologic and neuropsychologic functions. PMID- 11022859 TI - The usefulness of rabbits as an animal model for the neuropathological assessment of neurotoxicity following the administration of vincristine. AB - Vincristine is an effective chemotherapeutic agent for a variety of human neoplasms, but has dose-limiting neurotoxicity. Since laboratory rodents have proven to be refractive in such neurotoxicological studies, we conducted a neuropathological and behavioral assessment in rabbits treated with vincristine at doses known to be both chemotherapeutically effective and neurotoxic in humans. Rabbits (Kbl: NZW) were given vincristine intravenously at doses of 0 (saline), 200, 250 or 300 microg/kg once a week for 6 weeks, 500 microg/kg once a week for 3 weeks, or a single 500 microg/kg administration. Detailed periodic neurologic examination revealed ataxia in a few animals. Pathologically, axonal injury progressing to fiber degeneration was observed in sensory tracts such as the posterior spinocerebellar tract and posterior funiculus, and in peripheral nerves after treatment with vincristine. These alterations were observed even after a single dose of 500 microg/kg. In the group given weekly doses of 500 microg/kg, neuronal chromatolysis was also found in the spinal cord. These results suggest the rabbit is responsive to vincristine neurotoxicity producing a predominantly sensory neuropathy and confirming earlier studies. PMID- 11022860 TI - Schedule dependency of paclitaxel-induced neuropathy in mice: a morphological study. AB - Paclitaxel, which is known to induce peripheral neuropathy in humans, was administered to BDF1 mice, and a neuropathological examination was then conducted. Paclitaxel was administered at a dose of 30 mg/kg once or several times at different intervals, 3 times, every 3 hours (q3hx3), every day (q1dx3), every 2 days (q2dx3) and 4 times, once a week (q7dx4). The spinal cord, spinal ganglion and peripheral nerves (sciatic and tibial) were then processed for neuropathological examination following perfusion of the anesthetized mice. The effect on the peripheral nerves was evaluated by counting the number of abnormal nerve fibers in the preparations of teased nerve fibers and in the epoxy resin 1 microm-thick sections. The drug-induced degeneration, such as axonal and myelin fragmentations and phagocytosis in the nerve fibers, was detected in the dorsal funiculus, the dorsal spinal roots and the peripheral nerves (sciatic and tibial). No drug-related degeneration was observed in the motor neurons in the spinal cord nor in the ventral spinal roots. The single treatment did not induce any changes. The severity of the degeneration was as follows: q2dx3>q1dx3>>q3hx3 and q7dx4. The degeneration in the mice treated on q3hx3 and q7dx4 was very slight, but it was clear that paclitaxel also induced degeneration on these schedules. These results suggest that paclitaxel induces a predominantly sensory neuropathy in mice and the severity is obviously dependent on the treatment schedule. PMID- 11022861 TI - Effects of atropine sulphate on seizure activity and brain damage produced by soman in guinea-pigs: ECoG correlates of neuropathology. AB - The present study describes the effects of pyridostigmine (PYR; 0.2 mg/kg) and atropine sulphate (AS; 5 mg/kg) on guinea-pigs intoxicated by a high dose (2xLD50) of the organophosphate compound, soman, an irreversible inhibitor of acetylcholinesterase. The medication was shown to counteract the acute respiratory distress and lethality normally produced by the intoxication. Moreover, due to the central activity of AS, soman-induced electrocorticographic (ECoG) seizure activity was either totally prevented, or reduced in duration and overall intensity. In addition, as established in the 24-hr survivors, seizure related neuropathology was either prevented, or reduced in topographical extent and severity. An attempt to correlate our electrographic and morphological findings gives evidence that (a), the occurrence of seizure activity is the primary factor necessary for the development of acute neuropathology; (b), the duration of ECoG seizures is a secondary factor, on which the topographical distribution of brain damage finally depends; (c), the minimal duration of seizures necessary to produce 24 hr-damage in the most sensitive areas (e.g. the amygdala) is less than 70 min; (d), the overall intensity/power of epileptiform discharges is a tertiary factor which influences the severity of damage; (e), in addition, ECoG power spectral analysis suggested that an acute increase of relative power in the lower (delta) frequency band might be a real-time external marker of the starting cerebral lesions and is thus predictive for their future installation. All these data confirm the tight relationships which exist between seizure activity and neuropathology in soman poisoning, and suggest that refined, standardized analysis of electrographic parameters drawn from ECoG tracings and power spectrum might serve as a useful tool to predict the presence, localization, and severity of soman-induced brain damage. PMID- 11022862 TI - Muscarinic receptor-mediated pyridostigmine-induced neuronal apoptosis. AB - Pyridostigmine is a reversible cholinesterase (ChE) inhibitor that is associated with neurologic dysfunction involving both central and peripheral nervous systems. To determine the neurotoxic potential of pyridostigmine, rats were sacrificed at intervals after drug administration (0.5-1.85 mg/kg, i.p., twice daily for 4 days) and brains examined histologically. ChE inhibition was used as a biomarker of pyridostigmine activity. Using the in situ terminal deoxynucleotidyl transferase nick-end labeling of DNA fragments (TUNEL) method and electron microscopy, apoptotic brain cell death was noted in cerebral cortex over a dose range of 0.5-1.85 mg/kg and at the higher dose (1.85 mg/kg), apoptosis was also noted in striatum and hippocampus. These responses were blocked by pretreatment with atropine. Rat cortical cells in culture also underwent apoptosis when exposed to pyridostigmine (250 microM for 24 hr), indicating that the pyridostigmine can initiate apoptosis, independent of peripheral mechanisms. Pretreatment of cells with atropine (10 microM) inhibited pyridostigmine-induced apoptosis, confirming the response was mediated by muscarinic receptors. Short term treatment of rats with pyridostigmine (1.85 mg/kg twice daily for 4 days) induced a prolonged apoptotic response, which was evident in rat cortex up to 30 days after the last dose. Active apoptosis persisted, despite recovery of serum ChE activity. These in vivo and in vitro observations indicate that pyridostigmine can initiate a prolonged neurodegeneration. PMID- 11022863 TI - Dietary cadmium exposure alters characteristics of training, substitution, and tolerance when morphine is used as a discriminative stimulus. AB - This study examined the possibility that cadmium, a toxicant in high concentration in all tobacco products, may alter the stimulus properties of morphine. Adult male rats were exposed to regular laboratory chow (Group Control) or chow containing 100 ppm added cadmium chloride (Group Cadmium). Following an initial 30 day exposure period, control and cadmium-exposed animals were trained to discriminate between i.p. injections of 3.00 mg/kg morphine sulfate and vehicle (distilled water) in a two-choice drug discrimination task. Subsequently, the morphine dose-effect generalization function (0.75-6.00 mg/kg) was determined for control and cadmium-exposed animals. Additional substitution tests were conducted with increasing doses of the high efficacy mu agonist fentanyl (0.0016 0.04 mg/kg), the intermediate efficacy mu agonist (-)-metazocine (0.60-5.00 mg/kg), and the kappa agonist (+/-)-bremazocine (0.03-0.12 mg/kg). Also, increasing doses of the selective mu antagonist naloxone (0.0008-0.50 mg/kg) were presented against the training dose of morphine (3.00 mg/kg) and 0.02 mg/kg fentanyl. Finally, training was discontinued, and control and cadmium-exposed animals were injected with 8.00 mg/kg morphine in the home cage every 12 hr for 2 weeks, prior to redetermining the morphine dose effect function. Following a 1 week recovery period where morphine injections were discontinued, a final determination of the morphine dose-effect function was made. The results of the investigation indicated that cadmium exposure, without affecting the rate changing properties of the drugs, slowed initial acquisition of the morphine discrimination, decreased the potency of selective doses of naloxone with respect to antagonizing the stimulus effects of morphine and fentanyl, and blocked the development of tolerance to morphine. Morphine, fentanyl, and (-)-metazocine generalized (substituted) equally across both groups, while (+/-)-bremazocine failed to substitute for the morphine stimulus in either group. These findings add to the growing literature on the interaction between metal poisoning and drug selection/abuse. PMID- 11022864 TI - Astrocytes in methylmercury, ammonia, methionine sulfoximine and alcohol-induced neurotoxicity. AB - Astrocytes occupy approximately 25% of the CNS volume. Their "foot" processes are closely associated with synapses, nodes of Ranvier, axonal tracts, and capillaries. Astrocytic functions include neurotrophic factor secretion, control of extracellular pH, inactivation of glutamate, as well as uptake and metabolism of neurotransmitters. Astrocyte-neuron interactions provide strategic sites for actions of numerous chemical compounds. In this manuscript, we discuss examples of toxins that directly affect astrocyte function (methylmercury, ammonia, methionine sulfoximine, and alcohol), leading to an altered homeostatic control of the extracellular milieu and neuronal dysfunction. In addition, the potential role of astrocytic proteins, the metallothioneins, in attenuating the neurotoxicity of methylmercury is discussed. PMID- 11022865 TI - The PON1 gene and detoxication. AB - It has been assumed since its discovery that serum paraoxonase (PON1) plays a major role in the detoxication of specific organophosphorus compounds. It was also assumed that individuals with low PON1 activity would be more susceptible to paraoxon/parathion poisoning than individuals with higher PON1 activity. Evidence supporting this hypothesis was provided by injection of rabbit PON1 into rodents. Injected PON1 protected against paraoxon toxicity in rats and chlorpyrifos oxon toxicity in mice. The recent availability of PON1 knockout mice has provided an in vivo system with which one can more closely examine the role of PON1 in detoxication. PON1 knockout mice demonstrated dramatically increased sensitivity to chlorpyrifos oxon and diazoxon and moderately increased sensitivity to the respective parent compounds. The PON1 knockout mutation also resulted in the elimination of liver PON1 activity, accounting for the dramatic increase in sensitivity to chlorpyrifos oxon and diazoxon. Totally unexpected was our finding that the PON1 knockout mice were not more sensitive to paraoxon. This was particularly surprising in light of the earlier enzyme injection experiments. Differences in the relative catalytic efficiencies of rabbit vs. mouse PON1 for the specific oxon forms explain these observations. Mouse PON1 has good catalytic efficiency for the hydrolysis of diazoxon and chlorpyrifos oxon, but a poor efficiency for paraoxon hydrolysis relative to rabbit PON1. The human PON1Q192 isoform has a catalytic efficiency similar to that of mice, whereas the human PON1R192 isoform has a much better catalytic efficiency, predicting that individuals expressing high levels of the PONIR192 isoform may have increased resistance to paraoxon toxicity. PMID- 11022866 TI - Multiple chemical sensitivity (MCS)--differential diagnosis in clinical neurotoxicology: a German perspective. AB - The multiple chemical sensitivity syndrome (MCS) is a new cluster of environmental symptoms which have been described and commented on for more than 15 years now in the USA. In the meantime it has also been observed in European countries. The main features of this syndrome are: multiple symptoms in multiple organ systems, precipitated by a variety of chemical substances with relapses and exacerbation under certain conditions when exposed to very low levels which do not affect the population at large. There are no lab markers or specific investigative findings. In our view, MCS is not a separate clinical syndrome but a collective term. A very small part of the patients in question may actually exhibit a somatic or psychosomatic response to low levels of a variety of chemicals in the environment. For another part, even if the MCS symptoms are induced by chemical substances in the environment, the basic hypersensitivity is a psychological stress reaction. In the third and largest group, the patients have been misdiagnosed, i.e. a somatic or psychiatric disease has been overlooked. There is a fourth group of patients in whom there is no evidence of any exposure at all but instead a belief system installed by certain physicians, the media and other groups in society. This paper tries to describe the neurological and neurotoxic aspects of MCS problems and to illustrate it with examples of an alleged outbreak of chronic neurotoxic disease caused by pyrethroids in Germany. Research strategy should establish clearly determined diagnostic criteria, agreement on the use of specific questionnaires as well as clinical and technical diagnostic procedures, prospective clinical studies of MCS patients and comparative groups as well as experimental approaches. PMID- 11022867 TI - Use of aggregating brain cell cultures to study developmental effects of organophosphorus insecticides. AB - Aggregating brain cell cultures of fetal rat telencephalon can be grown in a chemically defined medium for extended periods of time. After a phase of intense mitotic activity, these three-dimensional cell cultures undergo extensive morphological differentiation, including synaptogenesis and myelination. To study the developmental toxicity of organophosphorus compounds (OP), aggregating brain cell cultures were treated with parathion. Protein content and cell type-specific enzyme activities were not affected up to a concentration of 10(5) M. Gliosis, characterized by an increased staining for glial fibrillary acidic protein (GFAP), was observed in immature and in differentiated cells. In contrast, uridine incorporation and myelin basic protein (MBP) immunoreactivity revealed strong differences in sensitivity between these two developmental stages. These results are in agreement with the view that in vivo the development-dependent toxicity is not only due to changes in hepatic detoxification, but also to age related modifications in the susceptibility of the different populations of brain cells. Furthermore, they underline the usefulness of histotypic culture systems with a high developmental potential, such as aggregating brain cell cultures, and stress the importance of applying a large range of criteria for testing the developmental toxicity of potential neurotoxicants. PMID- 11022868 TI - Styrene-induced changes in amacrine retinal cells: an experimental study in the rat. AB - Dopamine (DA) is synthesized in amacrine cells and released upon membrane depolarization in a calcium-dependent way. Thus, it is recognized to function as a major neurotransmitter or modulator in vertebrate retina. Owing to DA modulating activity on cone-horizontal cells transmission, depletion or dysfunction of amacrine cells could interfere with chromatic processing, accounting for the acquired dyschromatopsia described among styrene-exposed workers. The present study has been designed to test the hypothesis that amacrine cells represent a vulnerable target of styrene in subchronically exposed rats. Ten female Sprague-Dawley rats were exposed to 300 ppm styrene 6 h/day, 5 days/week, for 12 weeks; ten rats exposed to fresh air served as a control group. Whole mounted retinas were used for the morphometry of tyrosine hydroxylase (TH) immunoreactive cells (IR). DA content and TH activity were measured by HPLC and electrochemical detection and glutathione (GSH) was measured by HPLC tandem mass spectrometry (LC-MS/MS). In treated rats, morphometric analysis showed a loss of TH-IR amacrine cells (6.2/mm2 vs. 8.7/mm2 recorded in controls, p = 0.002), without any peripheral-central variation in cell loss. DA content was also lower in exposed, as compared to control animals (208.64 vs. 267.98 microg/g w.w., p = 0.004). The activity of TH in the whole retina was similar in styrene-exposed and control rats when expressed as a function of the wet weight, whereas it was much higher in styrene-exposed rats (+64%) when expressed as a function of the number of TH-IR amacrine cells (p < 0.001). Finally, retinal GSH was reduced by 30% in exposed as compared to control rats (p = 0.01). In summary, retinal TH-IR cells were sensitive to styrene exposure, which seems to cause both structural and functional changes, represented by cell loss and DA depletion, respectively. These findings confirm the vulnerability of dopaminergic systems to styrene toxicity, providing some insights on the possible mechanism of loss in chromatic discrimination recorded among workers occupationally-exposed to styrene. PMID- 11022869 TI - Role of antioxidant lycopene in cancer and heart disease. AB - Lycopene, a carotenoid without provitamin-A activity, is present in many fruits and vegetables; however, tomatoes and processed tomato products constitute the major source of lycopene in North American diet. Among the carotenoids, lycopene is a major component found in the serum and other tissues. Dietary intakes of tomatoes and tomato products containing lycopene have been shown to be associated with decreased risk of chronic diseases such as cancer and cardiovascular diseases in several recent studies. Serum and tissue lycopene levels have also been inversely related with the chronic disease risk. Although the antioxidant properties of lycopene are thought to be primarily responsible for its beneficial properties, evidence is accumulating to suggest other mechanisms such as modulation of intercellular gap junction communication, hormonal and immune system and metabolic pathways may also be involved. This review summarizes the background information about lycopene and presents the most current knowledge with respect to its role in human health and disease. PMID- 11022870 TI - Use of subjective global assessment to identify nutrition-associated complications and death in geriatric long-term care facility residents. AB - OBJECTIVE: The primary objective of this study was to assess the use of Subjective Global Assessment to identify nutrition-associated complications and death in a geriatric population. A secondary objective was to evaluate the ability of Subjective Global Assessment to identify geriatric residents of long term care facilities who were undernourished or at risk for developing undernutrition. METHODS: Fifty-three consecutive residents who were > or = 65 years of age and had been residing in a long-term care facility for < 2 weeks were enrolled in the study. The Subjective Global Assessment Classification technique was performed according to the procedure outlined by Detsky and colleagues. Residents were classified as well-nourished (A), mild/moderately undernourished (B) or severely undernourished (C). In addition, a Subjective Global Assessment Composite Score was derived. Subjective Global Assessment measures were compared with two traditional objective measurements of nutritional status: serum albumin and serum total cholesterol. Outcome measurements of nutrition-associated complications were determined over a 3-month period by recording the incidence of major infections, decubitus ulcers, nutrition-related hospital readmissions, and mortality. RESULTS: Sixteen residents (30.2%) were categorized as Subjective Global Assessment class A, 28 residents (52.8%) were class B, and 9 residents (17%) were class C. A significant association was found between nutritional status as determined by Subjective Global Assessment Composite Score and nutrition-associated complications (p<0.05). Subjective Global Assessment Classification was related to death (p<0.05) with severely undernourished residents having the highest mortality rate. Hypoalbuminemia only demonstrated a significant relationship with nutrition-associated complications (p<0.05), whereas hypocholesterolemia was associated with death (p<0.05). CONCLUSIONS: Subjective Global Assessment of nutritional status appears to be a simple, noninvasive and cost-effective tool for assessing nutritional status of geriatric residents in long-term care facilities. This assessment tool is also beneficial for identifying patients with increased risk of nutrition-associated complications as well as death. PMID- 11022871 TI - Health advantages and disadvantages of weight-reducing diets: a computer analysis and critical review. AB - BACKGROUND: Some weight-loss diets are nutritionally sound and consistent with recommendations for healthy eating while others are "fad" diets encouraging irrational and, sometimes, unsafe practices. OBJECTIVE: The purpose of the study was to compare several weight loss diets and assess their potential long-term effects. DESIGN: Eight popular weight-loss diets were selected (Atkins, Protein Power, Sugar Busters, Zone, ADA Exchange, High-Fiber Fitness, Pritikin and Omish) to be non-clinically analyzed by means of a computer to predict their relative benefits/potential harm. A summary description, menu plan and recommended snacks were developed for each diet. The nutrient composition of each diet was determined using computer software, and a Food Pyramid Score was calculated to compare diets. The Mensink, Hegsted and other formulae were applied to estimate coronary heart disease risk factors. RESULTS: Higher fat diets are higher in saturated fats and cholesterol than current dietary guidelines and their long term use would increase serum cholesterol levels and risk for CHD. Diets restricted in sugar intake would lower serum cholesterol levels and long-term risk for CHD; however, higher carbohydrate, higher fiber, lower fat diets would have the greatest effect in decreasing serum cholesterol concentrations and risk of CHD. CONCLUSIONS: While high fat diets may promote short-term weight loss, the potential hazards for worsening risk for progression of atherosclerosis override the short-term benefits. Individuals derive the greatest health benefits from diets low in saturated fat and high in carbohydrate and fiber: these increase sensitivity to insulin and lower risk for CHD. PMID- 11022872 TI - The effect of caffeinated, non-caffeinated, caloric and non-caloric beverages on hydration. AB - OBJECTIVE: To examine the effect of various combinations of beverages on hydration status in healthy free-living adult males. METHODS: In a counterbalanced, crossover manner, 18 healthy adult males ages 24 to 39, on four separate occasions, consumed water or water plus varying combinations of beverages. Clinical guidelines were used to determine the fluid allowance for each subject. The beverages were carbonated, caffeinated caloric and non-caloric colas and coffee. Ten of the 18 subjects consumed water and carbonated, non caffeinated, citrus soft drink during a fifth trial. Body weight, urine and blood assays were measured before and after each treatment. RESULTS: Slight body weight loss was observed on all treatments, with an average of 0.30% for all treatments. No differences (p>0.05) among treatments were found for body weight changes or any of the biochemical assays. Biochemical assays conducted on first voids and 24 hour urines included electrolytes, creatine, osmolality and specific gravity. Blood samples were analyzed for hemoglobin, hematocrit. electrolytes, osmolality, urea nitrogen, creatinine and protein. CONCLUSIONS: This preliminary study found no significant differences in the effect of various combinations of beverages on hydration status of healthy adult males. Advising people to disregard caffeinated beverages as part of the daily fluid intake is not substantiated by the results of this study. The across-treatment weight loss observed, when combined with data on fluid-disease relationships, suggests that optimal fluid intake may be higher than common recommendations. Further research is needed to confirm these results and to explore optimal fluid intake for healthy individuals. PMID- 11022873 TI - Comparative cholesterol lowering properties of vegetable oils: beyond fatty acids. AB - OBJECTIVE: Our laboratory has previously reported that the hypolipidemic effect of rice bran oil (RBO) is not entirely explained by its fatty acid composition. Although RBO has up to three times more serum cholesterol-raising saturated fatty acids (SATS) than some unsaturated vegetable oils, we hypothesized that its greater content of the unsaponifiables would compensate for its high SATS and yield comparable cholesterol-lowering properties to other vegetable oils with less SATS. METHODS: To study the comparative effects of different unsaturated vegetable oils on serum lipoprotein levels, nine cynomologus monkeys (Macaca fascicularis) were fed diets, for four weeks, in a Latin square design, containing rice bran, canola or corn oils (as 20% of energy) in a basal mixture of other fats to yield a final dietary fat concentration of 30% of energy. All animals were fed a baseline diet containing 36% of energy as fat with 15% SATS, 15% monounsaturated fatty acids (MONOS) and 6% polyunsaturated fatty acids (POLYS). RESULTS: Despite the lower SATS and higher MONOS content of canola oil and the higher POLYS content of corn oil, RBO produced similar reductions in serum total cholesterol (TC) (-25%) and low density lipoprotein cholesterol (LDL C) (-30%). In addition, as compared to the baseline diet, the reduction in serum TC and LDL-C cholesterol with RBO was not accompanied by reductions in high density lipoprotein cholesterol (HDL-C) which occurred with the other two dietary oils. Using predictive equations developed from data gathered from several studies with non-human primates, we noted that the observed serum TC and LDL-C lowering capabilities of the RBO diet were in excess of those predicted based on the fatty acid composition of RBO. CONCLUSIONS: These studies suggest that non fatty acid components (unsaponifiables) of RBO can contribute significantly to its cholesterol-lowering capability. PMID- 11022874 TI - Homocysteine elevation in sickle cell disease. AB - OBJECTIVE: Ischemic complications are common in patients with sickle cell disease. Hyperhomocysteinemia is a risk factor for arteriosclerosis and venous thrombosis, and given the propensity of patients with sickle cell disease to develop ischemic complications, we hypothesized that they might have elevated plasma homocysteine concentrations. METHODS: Plasma concentrations of homocysteine, vitamin B12 and folate were measured in 49 adults with sickle cell disease and 16 normotensive Black controls. All subjects with sickle cell disease had been prescribed folic acid 1 mg by mouth daily. RESULTS: The median plasma concentration of homocysteine of subjects with sickle cell disease was approximately 1.5-fold higher than that of controls (p=0.0008). This difference persisted, even when subjects with renal insufficiency were excluded. Plasma folate levels were 1.5-fold higher in subjects with sickle cell disease than in controls (p=0.0498). There was no significant difference in plasma vitamin B12 concentrations between the two groups. There was no difference in plasma homocysteine concentrations between transfused and non-transfused sickle cell subjects. CONCLUSIONS: Patients with sickle cell disease have elevated plasma concentrations of homocysteine in spite of elevated plasma folate levels and vitamin B12 concentrations similar to those observed in controls. Based on these data, we hypothesize that the concentration of folate required to normalize plasma homocysteine levels in patients with sickle cell disease may be higher than that of normal controls and that patients with sickle cell disease have a higher nutritional requirement for folic acid than the general population. PMID- 11022875 TI - The effects of a multivitamin/mineral supplement on micronutrient status, antioxidant capacity and cytokine production in healthy older adults consuming a fortified diet. AB - BACKGROUND: Inadequate micronutrient intake among older adults is common despite the increased prevalence of fortified/enriched foods in the American diet. Although many older adults take multivitamin supplements in an effort to compensate, studies examining the benefits of this behavior are absent. OBJECTIVE: To determine whether a daily multivitamin/mineral supplement can improve micronutrient status, plasma antioxidant capacity and cytokine production in healthy, free-living older adults already consuming a fortified diet. METHODS: An eight-week double-blind, placebo-controlled clinical trial among 80 adults aged 50 to 87 years (mean = 66.5 +/- 8.6 years). RESULTS: Multivitamin treatment significantly increased (p<0.01, compared to placebo) plasma concentrations of vitamins D (77 to 100 nmol/L), E (27 to 32 micromol/L), pyridoxal phosphate (55.1 to 75.2 nmol/L), folate (23 to 33 nmol/L), B12 (286 to 326 pmol/L)), C (55 to 71 micromol/L), and improved the riboflavin activity coefficient (1.23 to 1.15), but not vitamins A and thiamin. The multivitamin reduced the prevalence of suboptimal plasma levels of vitamins E (p=0.003), B12 (p=0.004), and C (p=0.08). Neither glutathione peroxidase activity nor antioxidant capacity (ORAC) were affected. No changes were observed in interleukin-2, -6 or -10 and prostaglandin E2, proxy measures of immune responses. CONCLUSIONS: Supplementation with a multivitamin formulated at about 100% Daily Value can decrease the prevalence of suboptimal vitamin status in older adults and improve their micronutrient status to levels associated with reduced risk for several chronic diseases. PMID- 11022876 TI - Vegetarianism and ischemic heart disease in older Chinese women. AB - OBJECTIVE: This study compared the risk of ischemic heart disease among older vegetarian Chinese women with that of older non-vegetarian women. METHODS: 90 vegetarian Chinese women over 70 years old in Hong Kong were screened for ischemic heart disease by electrocardiogram (ECG) and cardiovascular questionnaire. They were compared with 90 non-vegetarian women of similar age examined in a previous local survey. The effects of confounding factors were adjusted by stepwise logistic regression analysis. RESULTS: The percentages of subjects with ischemic heart disease defined by symptoms and ECG or by ECG alone were significantly lower in vegetarian women (p <0.005 and p<0.05 respectively). Vegetarians had lower serum cholesterol levels: more were old age home residents and were less likely to perform regular exercise. On stepwise logistic regression, using probable ischemic heart disease defined by questionnaire and ECG as outcome measure, vegetarianism was the only significant predictor (OR 0.3, 95%CI 0.1-0.6, p<0.005). CONCLUSIONS: Vegetarian older Chinese women had lower risk of ischemic heart disease when compared with non-vegetarians. Apart from lower serum cholesterol levels, vegetarianism may have other protective factors against ischemic heart disease. PMID- 11022877 TI - Older adults need guidance to meet nutritional recommendations. AB - OBJECTIVE: The purpose of this study was to compare the diet of healthy, free living senior volunteers to the dietary reference intakes (DRIs) and Food Guide Pyramid recommendations. METHODS: This study was a cross-sectional assessment of dietary habits, as measured using a standardized food frequency questionnaire, among 1,740 healthy Southwestern U.S. adults, aged 51 to 85 years. Assessment of independently-living volunteers to chemoprevention trials provides an efficient mechanism to profile typical dietary habits among the older adult population. RESULTS: Daily estimated macronutrient intakes exceeded recommended proportions of protein and fat. In contrast, more than 60% of this senior population reported dietary vitamin D, vitamin E, folate and calcium intakes below estimated average requirements (EAR). Based on the Food Guide Pyramid recommendations, fewer than 10% of the older adults consumed the recommended daily dairy and grain servings. More females than males consumed recommended vegetable (49% versus 40%) and fruit (53% versus 48%) servings (p < 0.05). More males consumed recommended grain (11% versus 7%) and protein (78% versus 73%) servings (p < 0.05) than females. CONCLUSIONS: Mean micronutrient intakes compared well with DRIs, although fewer than one-half of these older adults consumed recommended levels for vitamin D, vitamin E, folate, and calcium or daily food servings of dairy, grains, vegetables or fruits. Since the beneficial aspects of foods are not limited to essential nutrients, nutrition recommendations to older adults may be improved by emphasizing daily servings of nutrient-dense choices within the Food Pyramid. PMID- 11022879 TI - The effects of signal salience and caffeine on performance, workload, and stress in an abbreviated vigilance task. AB - In 2 experiments, a 12-min computerized vigilance task was demonstrated to reproduce the vigilance decrement, high workload (NASA-TLX), and stressful character (Dundee Stress State Questionnaire) of vigilance tasks lasting 30 min or more. In Experiment 1, the abbreviated task was also shown to duplicate the signal salience effect, a major finding associated with long-duration vigilance tasks. Moreover, Experiment 2 showed that performance on the abbreviated task can be enhanced by caffeine - a drug that benefits long-duration tasks. This enhancement effect was limited to performance, however, suggesting that caffeine influences factors that control signal detection but not those that control task induced stress. The results parallel those obtained with long-duration tasks and support a resource-depletion model of the vigilance decrement. The abbreviated task might be useful in situations in which long-duration tasks are precluded (e.g., performance assessment batteries, neuropsychological testing, and brain imaging). PMID- 11022878 TI - A survey of World Wide Web use in middle-aged and older adults. AB - We conducted a survey to document World Wide Web use patterns in middle-aged (ages 40-59), young-old (ages 60-74), and old-old adults (ages 75-92). We conducted this survey of 550 adults 40 years old and over in southeastern Michigan, and the overall response rate was approximately 71%. The results suggested that (a) there are distinct age and demographic differences in individuals who use the Web; (b) middle-aged and older Web users are similar in their use patterns; (c) the two primary predictors for not using the Web are lack of access to a computer and lack of knowledge about the Web; (d) old-old adults have the least interest in using the Web compared with middle-aged and young-old adults; and (e) the primary content areas in learning how to use the Web are learning how to use electronic mail and accessing health information and information about traveling for pleasure. This research may serve as a preliminary attempt to ascertain the issues that must be faced in order to increase use of the World Wide Web in middle-aged and older adults. PMID- 11022880 TI - Assessment of the relationship between box weight and trunk kinematics: does a reduction in box weight necessarily correspond to a decrease in spinal loading? AB - Typically, the simplest and most cost-efficient ergonomic solution to offset the rising costs of low back injuries is to reduce the box weight that is lifted. However, there is limited research on how a worker interacts with the box. In the present study, we quantify the utility of reducing the weight that is lifted - specifically, how changes in the box weight affect trunk kinematics, trunk moments, and ultimately, spinal loads. In the experiment, 15 participants lifted a variety of box weights (from 9.1 to 41.7 kg) from knee height, carried it a distance of 5 feet (1.5 m), and placed it on a shelf at elbow height. For the lower weights, small increases in box weight (3-9 kg) were offset by the trunk dynamics (sagittal velocity), resulting in no difference in spinal loads. At the same time, spinal loads were found to be significantly higher for weights above 25 kg. Thus, when making ergonomic changes (reduction of box weight), it is important to consider how workers will interact with the box. These results indicate that purely weight-based ergonomic controls might not sufficiently reduce the risk of low back disorders. Furthermore, this study provides additional evidence of the utility of using more complex spinal load models (dynamic, multiple muscle models) when evaluating highly dynamic and complex tasks. PMID- 11022881 TI - Continuous assessment of back stress (CABS): a new method to quantify low-back stress in jobs with variable biomechanical demands. AB - Jobs with a high degree of variability in manual materials handling requirements expose limitations in current low-back injury risk assessment tools and emphasize the need for a probabilistic representation of the biomechanical stress in order to quantify both acute and cumulative trauma risk. We developed a hybrid assessment methodology that employs established assessment tools and then represents their evaluations in a way that emphasizes the distributions of biomechanical stress. Construction work activities in the home building industry were evaluated because of the high degree of variability in the manual material handling requirements. Each task was evaluated using the Revised NIOSH Lifting Equation, The University of Michigan Three-Dimensional Static Strength Prediction Program, and the Ohio State University Lumbar Motion Monitor Model. The output from each model was presented as time-weighted histograms of low-back stress, and the assessments were compared. The results showed considerable differences in what were considered high-risk activities, indicating that these 3 assessment tools consider the risk of low-back injury from different perspectives. The time weighted distribution aspect of this methodology also contributed vital information toward the identification of high-risk activities. These results illustrate the necessity for more advanced low-back injury risk assessment techniques for jobs with highly variable manual materials handling requirements. PMID- 11022882 TI - Putting it all together: improving display integration in ecological displays. AB - Computer displays are being designed for increasingly larger industrial systems. As the application domain scales up, maintaining integration across different kinds of views becomes more challenging. This paper presents the results of a study of three different approaches to integration based on the spatial and temporal proximity of related information objects. The domain used for evaluation was a simulation of an industry-scale conventional power plant. All three displays were ecological displays developed using an abstraction hierarchy analysis. Views were integrated in a high-space/low-time, low-space/high-time, and high-space/high-time integration of means-end related objects. During a fault detection and diagnosis task, it was found that a low level of integration, high space/ low-time, provided the fastest fault detection time. However, the most integrated condition, high-space/high-time, resulted in the fastest and most accurate fault diagnosis performance. Actual or potential applications of this research include computer displays for large-scale systems such as network management or process control, for which problem solving is critical and integration must be maintained. PMID- 11022883 TI - Team mental models: techniques, methods, and analytic approaches. AB - Effective team functioning requires the existence of a shared or team mental model among members of a team. However, the best method for measuring team mental models is unclear. Methods reported vary in terms of how mental model content is elicited and analyzed or represented. We review the strengths and weaknesses of vatrious methods that have been used to elicit, represent, and analyze individual and team mental models and provide recommendations for method selection and development. We describe the nature of mental models and review techniques that have been used to elicit and represent them. We focus on a case study on selecting a method to examine team mental models in industry. The processes involved in the selection and development of an appropriate method for eliciting, representing, and analyzing team mental models are described. The criteria for method selection were (a) applicability to the problem under investigation; (b) practical considerations - suitability for collecting data from the targeted research sample; and (c) theoretical rationale - the assumption that associative networks in memory are a basis for the development of mental models. We provide an evaluation of the method matched to the research problem and make recommendations for future research. The practical applications of this research include the provision of a technique for analyzing team mental models in organizations, the development of methods and processes for eliciting a mental model from research participants in their normal work environment, and a survey of available methodologies for mental model research. PMID- 11022884 TI - The effects of task complexity and experience on learning and forgetting: a field study. AB - This paper examines the effects of task complexity and experience on parameters of individual learning and forgetting. Three attributes of task complexity and experience are addressed: the method, machine, and material employed. The task involved a high-manual-dexterity skill taken from an operating textile assembly plant; there were 2853 individual participant learning/forgetting episodes. A parametric model of individual learning and forgetting that allows the evaluation of worker response to the attributes of task complexity and experience is discussed. Results indicate that both task complexity and experience significantly affect learning and forgetting rates. Potential applications of this research include the allocation of workers to tasks based on individual learning/forgetting characteristics. PMID- 11022885 TI - An empirical comparison of alternative methodologies for the evaluation of configural displays. AB - Two different methodologies (visual, memory) were used to evaluate alternative versions of the same configural display. One version (composite display) had several graphical design techniques applied, whereas the other version (baseline display) did not. Two types of information probes (high-level property, low-level data) were administered. When the displays were visible during completion of the probes (visual methodology), the display manipulation had the largest impact on performance (composite display associated with better performance); when the displays were not visible (memory methodology) the probe manipulation had the largest impact on performance (high-level probes associated with better performance). These results are interpreted in light of the mutually interacting constraints introduced by factors in display design, task requirements, and the participants' cognitive and perceptual capabilities/limitations. Implications for both the design and the evaluation of displays and interfaces in general are discussed. Actual or potential applications of this research include design techniques for improving the quality of graphic displays and methodological insights for interpreting previous research and guiding future experimentation. PMID- 11022886 TI - Isoperformance and personnel decisions. AB - Isoperformance curves are contour lines or surfaces that trace all combinations of two or more determinants that suffice to produce a specified level of performance hence the term isoperformance. When the determinants are aptitude and training time or time on the job, an isoperformance curve allows one to identify individuals who have sufficient aptitude for the work that they can be expected to reach a specified standard of performance in a specified length of time. This paper discusses isoperformance in the personnel selection and classification literature and then develops and illustrates the uses of isoperformance curves in personnel decisions. The idea of aptitude sensitivity is introduced, and the relationship between this idea and predictive validity is explained. The paper also indicates how isoperformance can be used to detail trade-offs among training time, the minimum qualifying score, attrition rate, quotas, and the specified level of criterion performance, Isoperformance finds direct application in the multiple-job assignment problem, exemplified by the choice of which recruit to send to which training school in the military services. PMID- 11022887 TI - Is dual-task performance necessarily impaired in space? AB - Recent single-subject experiments in space have reported impaired dual-task performance that could result from either a direct effect of microgravity on the central nervous system or from the multistressor environment. We sought to distinguish between these hypotheses using 6 astronauts in the 16-day NASA Neurolab mission, testing them at intervals with a dual task consisting of primary pursuit tracking without vision of the hand and secondary reaction time (RT). The participants were highly trained, instructed to maintain a fixed attention strategy, and restrained in the apparatus. The results showed that absolute and variable tracking error, as well as correct RT and the standard deviation of RT, were unimpaired. However, RT errors became more variable, an effect attributed to a decrease in strategic control. We conclude that the impairments observed in previous dual-task space experiments can be attributed to stressors rather than to microgravity and that performance deficits are probably not a necessary concomitant of space flight if attention is paid to task design and astronaut training. PMID- 11022888 TI - Effect of aftermarket automobile window tinting films on driver vision. AB - This study was conducted to determine the level of automobile window tint that causes a significant reduction of vision for automobile drivers. Contrast sensitivity was measured on 20 participants, of whom 10 were age 20 to 29 years and 10 were age 60 to 69 years, through a stock automobile window (control) and two windows darkened with plastic film. For the younger drivers, a car window with 37% transmittance did not significantly reduce contrast sensitivity, but a darker tint of 18% transmittance reduced contrast sensitivity at higher spatial frequencies. For the older drivers, a tint of 37% transmittance significantly reduced mid-to- high spatial frequency contrast sensitivity. The typical state standard (no tint with less than 35% transmittance) would thus seem to be appropriate for younger drivers; however, further examination of the standard may be necessary in regard to older drivers. Actual or potential applications of this research include guidelines and regulations regarding tinting of automobile windows. PMID- 11022889 TI - The immediate effects of glare and electrochromic glare-reducing mirrors in simulated truck driving. AB - In this experiment 12 experienced truck drivers drove a fixed-base driving simulator for three 8-h sessions under simulated nighttime driving conditions. Sessions included (a) no glare, (b) intermittent glare presented in the exterior rearview mirrors to simulate following vehicles, and (c) intermittent glare with electrochromic glare reduction. The driving task combined vehicle control on straight and curved road segments with detection of pedestrians appearing alongside the road and targets appearing in the rearview mirrors. The presence of glare slowed detection of pedestrians and, to a lesser extent, slowed the detection of targets appearing in mirrors. Glare was also associated with increased lane position variability, reduced speed on curves, and, most consistently, increased steering variability. We found only meager evidence that electrochromic glare reduction improved target detection performance and no evidence that glare reduction improved vehicle control, despite the fact that participants consistently voiced positive preferences for glare reduction. The results will aid decision making that requires incorporation of the benefits of electrochromic glare-reducing mirrors. PMID- 11022890 TI - Development of ideas on renovascular hypertension. AB - Stephen Hales was the first to measure blood pressure directly in the horse (1733), and the definitive studies on human nephrins by Richard Bright followed much later (1836). The relation between high blood pressure and renal disease was established by Mahomed (1872). The discovery of renin and its possible link with Bright's disease was made by Tigerstedt and Bergman (1898), but only the experimental production of renal hypertension by Goldblatt and his colleagues (1934) led to the delineation of the role of the kidney in human hypertension by a wide variety of methods. PMID- 11022891 TI - The pathophysiology of renin release in renovascular hypertension. AB - Renovascular hypertension (RVH) results from occlusion of blood flow to either kidney, which stimulates renin release. Increased renin leads to a series of actions that rapidly leads to increased systemic blood pressure. Experimental renovascular hypertension is developed in animals by placement of a clip that occludes more than 50% of renal blood flow to that kidney. The major stimulus for renin release in renovascular hypertension is the severe drop in hydrostatic pressure in the afferent arteriole, the location of the juxtaglomerular renin secreting granular cells. The pressure drop changes the degree of stretch of these cells which leads to baroreceptor-mediated renin release. The level of renin released can be modified by sympathetic nerves and to a lesser degree by the macula densa. Several hormone or vasoactive agents may augment renin released during RVH, but nearly all are secondary to changes in the pressure receptor mechanism. PMID- 11022892 TI - Kidney angiotensin receptors and their role in renal pathophysiology. AB - A considerable amount of data have implicated angiotensin receptors (AT receptors) in the development and maintenance of essential hypertension and renovascular hypertension as well as in progressive renal pathologies. Inhibition of angiotensin II (Ang II) action by blocking Ang II formation through angiotensin-converting enzyme (ACE) inhibitors, or by blocking AT1 receptors directly using subtype-selective nonpeptide antagonists, has been found to attenuate the proteinuria, microalbuminuria, glomerulosclerosis, and nephrosclerosis in a variety of experimental models and in clinical trials. This review will first broadly discuss AT receptor subtypes in terms of their structure, function, tissue distribution and signaling. Secondly, the mechanistic differences between ACE inhibition and AT1 receptor blockade will be examined because these pharmaceutical agents are widely used tools to investigate the role of AT receptors in renal disease. Lastly, experimental models of essential hypertension, renovascular hypertension and progressive renal disease will be presented, which include the Fawn-hooded rat, the stroke prone spontaneously hypertensive rat, renal mass ablation and the 2K1C and 1K1C animal models. The overall goal of this review is to critically evaluate the data regarding the role of AT receptors in the pathophysiology of renal disease. PMID- 11022893 TI - Renal mechanisms of angiotensin II-induced hypertension. AB - The kidneys play a central role in the long-term control of arterial pressure by regulating sodium balance and extracellular fluid volume. The renin-angiotensin system is important in the regulation of the arterial pressure through its chronic effects on the pressure natriuresis relationship. Under physiologic conditions, angiotensin II (Ang II) is important in causing the long-term relationship between arterial pressure and sodium excretion to be very steep, so that minimal changes in arterial pressure are necessary to maintain sodium balance in response to variations in sodium intake. An inability to suppress Ang II formation in response to increases in sodium intake can lead to salt-sensitive hypertension. Excess formation of Ang II, such as in renovascular hypertension, causes the pressure natriuresis relationship to be shifted to higher arterial pressures so that higher arterial pressures are necessary to maintain sodium balance. Ang II decreases pressure natriuresis by enhancing tubular reabsorption and/or reducing glomerular filtration. Because Ang II does not decrease glomerular filtration in most circumstances, the sodium retaining actions of Ang II are usually caused by increased tubular reabsorption. However, there are a number of pathophysiologic conditions where Ang II interacts with various local autocrine and paracrine factors (such as nitric oxide [NO], eicosanoids, adenosine, and superoxide) to influence glomerular filtration rate. Ang II enhances tubular reabsorption either indirectly, through aldosterone stimulation, via alterations in renal hemodynamics (physical factors or medullary blood flow), or by directly enhancing tubular sodium transport. Converting enzyme inhibitors or Ang II receptor antagonists improve pressure natriuresis and are very effective in the treatment of various forms of hypertension associated with normal or enhanced activity of the renin-angiotensin system. PMID- 11022894 TI - Epidemiology and clinical presentation. AB - Renovascular disease appears to be increasing in prevalence, particularly in older subjects with atherosclerotic disease elsewhere. Its clinical manifestations and presentation are changing because of rapid advances in medical therapy and other comorbid events. Although fibromuscular dysplasia and other diseases affecting the renal artery can produce the syndrome of renovascular hypertension, atherosclerotic renal artery stenosis is the most common clinical entity. It can produce a spectrum of manifestations, ranging from asymptomatic ("incidental"), identified during angiographic evaluation of other conditions, to progressive hypertension to accelerated cardiovascular disease with pulmonary edema and advanced renal failure. With the widespread application of drugs which block the renin-angiotensin system, including angiotensin-converting enzyme inhibitors and angiotensin antagonists, many cases of renovascular hypertension remain unsuspected and never produce adverse effects. Clinicians need to be alert to the potential for disease progression, with the potential for total renal artery occlusion and/or loss of viable renal tissue. Selection of patients for renal revascularization depends on individual balance of risks and benefits regarding the likely outcomes regarding both improvements in blood pressure control and preservation of renal function. PMID- 11022895 TI - Functional testing: renin studies. AB - Static tests of plasma renin activity (PRA) or renal vein renins, even when assessed in relationship to dietary salt intake by renin-sodium profiling, are insufficiently sensitive for reliable screening for renovascular hypertension. Angiotensin converting enzyme inhibitors (ACEIs) stimulate PRA selectively in patients with renovascular hypertension. In large series, the captopril stimulated PRA test has a high sensitivity in excess of 90% where it is validated against a functional response of an improvement or cure in blood pressure after an intervention to correct renal artery stenosis. However, there are wide variations in the reported accuracy of this test between studies. In part, this may reflect an inappropriate use of a functional test to screen for an anatomical abnormality of renal artery stenosis. Both ACEI-stimulated PRA and ACEI-induced changes in the renogram likely reflect a similar fundamental abnormality in the poststenotic kidney: a selective reduction in the postglomerular vascular resistance leading to a sharp fall in the glomerular filtration rate (GFR). This fall in GFR is manifest as a delay in tubular transit of renogram tracers, leading to an abnormal ACEI renogram, and to a reduction in macula densa NaCl delivery, leading to an abnormal ACEI renogram, and to a reduction in macula densa NaCl delivery, leading to an exaggerated rise in PRA. Under protocol conditions, both the ACEI-PRA and the ACEI-renogram tests have sensitivities for the detection of functional renovascular hypertension of greater than 90%. PMID- 11022896 TI - Functional testing: ACEI renography. AB - Angiotensin-converting enzyme inhibition (ACEI) renography is the only imaging examination that tests directly for the presence of renovascular hypertension (RVH); other imaging examinations test only for the presence of renal artery stenosis (RAS). Consensus panels have recommended that ACEI renograms be interpreted as low, intermediate, or high probability for RVH. ACEI renography is highly accurate in patients with normal renal function and suspected RVH. In this patient population, the sensitivity and specificity of ACEI renography for RAS are approximately 90%; as an initial approach, angiography is not cost effective. Data from 10 studies evaluating cure or improvement in blood pressure in 291 patients undergoing revascularization showed the mean positive predictive value of ACEI renography to be 92%. When azotemic patients present with suspected RVH, as many as 50% of patients may have an intermediate probability ACEI renogram and the sensitivity of detecting RVH falls to approximately 80% even when intermediate and high probability tests are combined. PMID- 11022897 TI - Doppler and ultrasound methods for diagnosis. AB - With the availability of ultrasonic duplex scanning we now have a method of safely screening those patients who may be thought to have renal artery stenosis as the basis for hypertension or renal failure. Modern instrumentation combined with an experienced technologist makes this a reasonable and accurate screening test. One of its major advantages is that it can be used for repeat studies to document the outcome of any form of intervention designed to remove or bypass areas of stenosis. We no longer need to wonder if a reconstruction has been successful or not. Another great advantage is its role in documenting both the degree of narrowing and its effect on kidney size. It is now known, because of information obtained with this technology, that high-grade renal artery stenoses are accompanied by a decrease in kidney size. It is also possible to document disease progression with atherosclerotic involvement of the renal arteries. With more widespread use of this method, we will be able to obtain useful information on how renal artery disease should be treated and monitored. PMID- 11022898 TI - Magnetic resonance angiography of the kidney. AB - Magnetic resonance angiography (MRA) of the kidneys has become a clinical standard for detecting renal artery stenosis. This test is performed by injecting a bolus of a gadolinium chelate and scanning with a three-dimensional volumetric data collection sensitized to the T1 shortening effects of gadolinium. In addition to displaying the renal arterial anatomy, atherosclerosis within the aorta and iliac arteries is commonly depicted. MRA is a time efficient and safe test when compared with conventional arteriography. Gadolinium-enhanced MRA has proven to have a high sensitivity for detecting stenoses in main and accessory renal arteries. Although false-negative studies are rare, overestimation of the degree of renal stenosis is problematic and may lead to false-positive diagnosis. To some extent this tendency to overestimate stenoses can be compensated for by performing phase contrast MRA, a type of MRA based on accumulated phase differences. As with conventional angiography, MRA is still only an anatomic test which provides little information about the functional significance of a stenosis. It is highly accurate in determining the number of renal arteries, the size of the kidneys, and the presence of any anatomic variants. Ultimately, MRA, needs to be combined with a functional test similar in concept to captopril renography. This test, termed MR renography together with MRA may replace the current multimodalitiy approach to the work-up of renovascular hypertension. PMID- 11022899 TI - Novel noninvasive techniques for studying renal function in man. AB - Renal artery stenosis is a major cause of renovascular hypertension in humans, and may lead to ischemic nephropathy and end-stage renal disease. The mechanisms responsible for the progressive renal functional and structural alterations have not been fully elucidated, partly because of the lack of reliable, noninvasive techniques capable of quantifying renal regional hemodynamics and function distal to a stenosis in the renal artery. Novel imaging tools now enable quantification of concurrent intrarenal (cortical and medullary) hemodynamics, segmental nephron dynamics (intratubular transit times and fluid concentrations), and renal function in the intact kidney. Fast computed tomography (CT) scanners, such as electron beam CT, allow discrimination of subtle alterations in renal perfusion and segmental nephron function consequent to changes in renal perfusion pressure, both within and below the range of renal blood flow autoregulation. This technique provides an opportunity to define intrarenal perfusion patterns and function in animals and patients with renal artery stenosis, and may provide insight into the effects of chronic unilateral or bilateral renovascular disease on both the hypoperfused and contralateral kidneys. This methodology may thereby prove to be very useful in the evaluation of renal disease in general, and the renovascular hypertensive patient in particular. PMID- 11022901 TI - Renal vascular disease: medical management, angioplasty, and stenting. AB - In the absence of large, prospective, quality randomized trials, there remains tremendous debate concerning the optimal management of patients with renal vascular disease. This debate is compounded by the fact these patients do not represent a homogeneous group; different causes and presentations each carry a different prognosis and potential response to therapy. Therapeutic options include medical management, surgery, or percutaneous approaches (angioplasty or stenting). This review examines the results of observational studies of medical and percutaneous therapies for blood pressure control and preservation of renal function. Generally, in patients with fibromuscular disease, the results of percutaneous management are superior to medical therapy. Although these observational studies are difficult to compare, in patients with atheromatous disease, the results with interventional and medical therapy appear roughly similar. There have been three randomized prospective trials of routine angioplasty versus medical management. These trials show little advantage to interventional therapies in those patients whose blood pressure is well controlled with medication who do not show progression of renal insufficiency during medical management. Based on these data, this review outlines a potential management strategy that relies on an individualized risk benefit assessment. PMID- 11022900 TI - Prospective studies of diagnosis and intervention: the Dutch experience. AB - This prospective multicenter study included 1,205 patients, who were referred for difficult-to-treat hypertension or analysis of possible secondary hypertension. After a standardized selection protocol based on sharply defined drug-resistant hypertension or renal function impairment during angiotensin-converting enzyme inhibition, patients underwent renal scintigraphy and a captopril-renin challenge test. A set of clinical characteristics was also recorded. Sensitivity and specificity of renal scintigraphy for diagnosing renal artery stenosis were 0.72 and 0.90 and of the captopril-renin test 0.77 to 0.91 and 0.69 to 0.75 depending on the criterion used. The clinical characteristics were used to construct a clinical prediction rule for renal artery stenosis, which had a sensitivity of 0.68 and a specificity of 0.87 at a cut-off level of 30% predicted probability. However, with the prediction rule a sensitivity of 0.90 could be reached by performing arteriography only in patients with a predicted probability of stenosis of > or =10%, resulting in a considerable reduction of the number of arteriograms to be made. A diagnostic strategy is advocated starting with drug resistant hypertension and continuing to renal arteriography only in patients with increased probability of stenosis. Patients with atherosclerotic renal artery stenosis were then randomized to balloon angioplasty (n = 56) versus antihypertensive medication (n = 50). Three months after randomization 22 patients from the medication group underwent balloon angioplasty in second instance. In an intention-to-treat analysis, no difference in blood pressure was found between the groups after 3 months, nor after 12 months of follow-up, although there was a small medication-sparing effect of balloon angioplasty. The lack of a beneficial effect of balloon angioplasty compared with medication could not be attributed to the high stenosis recurrence rate after angioplasty, nor to the fact that the inclusion criterion was set at a stenosis level of > or =50% so that patients with relatively mild stenosis were also included. Renal function after angioplasty was slightly better in the angioplasty group than in the medication group, and improvement of the renal scintigram occurred more often after angioplasty. Apart from the treatment of patients with specific characteristics, the presented therapeutic approach starts with extending the antihypertensive drug therapy to control blood pressure. Only if blood pressure cannot be controlled or if renal function deteriorates, balloon angioplasty (with stent placement) is indicated. PMID- 11022902 TI - Ischemic nephropathy/azotemic renovascular disease. AB - Atherosclerotic renal vascular disease can impair kidney perfusion and lead to deterioration of kidney function. The mechanisms by which reversible tissue injury becomes irreversible are not yet certain, although multiple pathways for activation of inflammatory cytokines and tissue fibrosis have been identified. The clinical hallmark of this disorder is loss of glomerular filtration beyond renal artery stenosis affecting the entire renal mass, usually associated with progressive hypertension and fluid retention. Some investigators believe that 12% to 18% of patients reaching end-stage renal disease in western countries may have lost kidney function because of azotemic renovascular disease. This is an important disorder to identify, because reduction of arterial pressure from antihypertensive therapy may further reduce kidney perfusion. Although administration of angiotensin-converting enzyme (ACE) inhibitors and angiotensin II antagonists lead to functional loss of glomerular filtration rate (GFR) beyond a stenotic lesion because of the removal of efferent actions of angiotensin II, other antihypertensive agents reduce renal perfusion also. Restoration of renal blood flow by surgical or endovascular methods can prevent progressive disease and sometimes improves renal function. However, clinical series commonly indicate that some patients lose further kidney function after revascularization. This may be explained partly by undetected renal atheremboli or other toxicity related to vascular repair. Hence, selection of patients for renal revascularization requires careful consideration of comorbid disease risk and the balance of risks and benefits regarding progressive renal disease. Searching for better methods of identifying those individuals at risk for irreversible loss of renal function and who might benefit from vascular repair is a high research priority. PMID- 11022903 TI - Simultaneous determination of aceclofenac and diclofenac in human plasma by narrowbore HPLC using column-switching. AB - A fully automated narrowbore high performance liquid chromatography (HPLC) with column-switching was developed for the simultaneous determination of aceclofenac and diclofenac from human plasma samples. Plasma sample (100 microl) was directly introduced onto a Capcell Pak MF Ph-1 column (20 x 4 mm I.D.) where primary separation was occurred to remove proteins and concentrate target substances using acetonitrile potassium phosphate (pH 7, 0.1 M) (14:86, v/v). The drug molecules eluted from MF Ph-1 column were focused in an intermediate column (35 x 2 mm I.D.) by the valve switching step. The substances enriched in intermediate column were eluted and separated on the narrowbore phenyl hexyl column (100 x 2 mm I.D.) using acetonitrile-potassium phosphate (pH 7, 0.02M) (33:67, v/v) when the valve status was switched back to A position. The method showed excellent sensitivity (detection limit of 10 ng ml(-1)) with small volume of samples (100 microl), good precision and accuracy, and speed (total analysis time 17 min) without any loss in chromatographic efficiency. The response was linear (r2 > or = 0.999) over the concentration range of 50-10,000 ng ml(-1). PMID- 11022904 TI - Adsorptive stripping voltammetric determination of the antidepressant drug sulpiride. AB - The electrochemical behaviour of the antidepressant drug sulpiride (SP) at a hanging mercury drop electrode (HMDE) is investigated. Linear sweep cathodic stripping voltammetry (LSCSV) was used to determine sulpiride in the presence of 0.01 M sodium acetate medium pH 10.5 and 25 +/- 1 degrees C. Different parameters such as, supporting electrolyte, pH, accumulation potential, scan rate, accumulation time and ionic strength, were tested to optimize the conditions for the determination of SP. The adsorbed form is reduced irreversibly. The linear concentration range is from 2 x 10(-9) to 5 x 10(-8) M SP. Experimentally, 2 x 10(-9) M (0.68 ppb) with accumulation time 60 s can be determined successfully. Furthermore, a theoretical detection limit of 2 x 10(-10) M (0.068 ppb) Sp was calculated. The interferences of some metal ions, ascorbic acid and some amino acids were studied. The method was applied to the analysis of tablets and spiked urine, with recoveries of 104 +/- 3 and 101 + 3, and the relative standard deviation of 3.3 and 3.4%, respectively. PMID- 11022905 TI - Binding of ketoprofen enantiomers in various human albumin preparations. AB - Published data conflict with respect to the enantioselective protein binding parameters of R(-) and S(+) ketoprofen. We studied whether differences in experimental conditions used and/or presence of interfering compounds could provide a possible explanation for these discrepancies. Equilibrium dialysis, supported by ultrafiltration (67 mM Sorensen phosphate buffer pH 7.4, 580 microM HSA, 37 degrees C) allowed the characteristics of the binding sites to be determined according to Scatchard's analysis. (R) and (S)-ketoprofen concentrations were measured by HPLC. The free (R)-ketoprofen/free (S)-ketoprofen (F(R)/F(S)) concentration ratio was calculated. The effect of octanoic acid (OA) found in currently marketed intravenous HSA solutions, and hippuric acid (HA), on F(R)/F(S) concentration ratio was considered. Two classes of binding sites were characterized for both enantiomers. The free (S)-ketoprofen concentrations remained equal to those of the (R)-antipode at low concentrations of racemate (2 35 microg ml(-1)) indicating non-stereoselective albumin binding over the therapeutic range. From 35 microg ml(-1), the free (S)-ketoprofen concentrations were slighty greater than those of its antipode. Both OA and HA induced an increase of the free fraction of the enantiomers by a two-fold to a 15-fold order of magnitude. OA, but not HA, showed a more pronounced effect for the (S)-form leading to a marked decrease in F(R)/F(S) concentration ratio (0.61). Differences in HSA preparations used and/or the presence of interfering compounds may explain the variability in the reported protein binding characteristics of ketoprofen enantiomers. PMID- 11022907 TI - Determination of zopiclone in tablets by HPLC and UV-spectrophotometry. AB - Rapid, simple and accurate chromatographic (HPLC) and spectrophotometric methods for the determination of zopiclone in tablets were elaborated. Acetonitrile was found to be a suitable extraction solvent. The samples were chromatographed on Nova-Pak C18 column and UV detection at 304 nm. The elution was achieved isocratically with a mobile phase of 0.067 M phosphate buffer pH 7.95 - acetonitrile (55:45, v/v). Diazepam was applied as an internal standard. The method was validated for precision, linearity, accuracy and limit of detection. The recovery (mean +/- SD) in HPLC was 99.85 + 0.04% and in the UV spectrophotometry 100.08 +/- 0.09%. PMID- 11022906 TI - Furosemide assay in pharmaceuticals by Micellar liquid chromatography: study of the stability of the drug. AB - A simplified high-performance liquid chromatographic procedure is described for the determination of furosemide (4-chloro-N-furfuryl-5-sulphamoylanthranillic acid), which makes use of UV detection, a C18, reversed-phase column, and micellar mobile phases of sodium dodecyl sulphate (SDS) and 1-propanol at pH 3 buffered with phosphate system. The most adequate experimental conditions to handle furosemide solutions in the analytical laboratory are studied. The mixture of furosemide and its degradation products which are formed upon light exposition was resolved with a mobile phase of 0.04 M SDS-2% propanol. Separation of furosemide from its common impurities and the hydrolytic product, 4-chloro-5 sulphamoylanthranillic acid, was also possible. A mobile phase of larger elution strength, such as 0.06 M SDS-8%, propanol was preferred to assay furosemide in several dosage forms (tablets, capsules, injectables and drops). The validity of the procedure was checked by analysing 27 pharmaceuticals commercialised in several countries. The label claim percentages and coefficients of variation were in the 95-102% and 0.05-1.3% ranges, respectively. The results showed that the procedure is suitable for routine analysis of the diuretic. PMID- 11022908 TI - Rapid bioanalytical determination of dextromethorphan in canine plasma by dilute and-shoot preparation combined with one minute per sample LC-MS/MS analysis to optimize formulations for drug delivery. AB - The determination of dextromethorphan in canine plasma is used to demonstrate the high throughput bioanalytical approach of automated dilute-and-shoot (DAS) sample preparation followed by a 1 min isocratic liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. Dilute-and-shoot preparation is commonly used for the determination of drugs in several biological matrices such as urine and saliva, but is not typically used with plasma samples because the amount of protein present in plasma can lead to a variety of problems including column failure. As a result, plasma sample preparation usually removes protein by precipitation, extraction or filtration; however, the dilute-and-shoot approach solubilizes proteins throughout the chromatographic portion of the assay. The attributes of this approach are compared with a previously validated liquid/liquid extraction procedure for determination of dextromethorphan in plasma. Accuracy and precision of both methods are similar. The lower limit of quantitation (LLOQ) of the dilute-and-shoot approach is much higher at 2 ng/ml versus 5 pg/ml with the liquid/liquid extraction; however, the sample throughput of the preparation portion of the dilute-and-shoot approach is more than 50-fold greater. The ruggedness of the dilute-and-shoot method was thoroughly investigated because of the problems traditionally associated with the direct injection of diluted plasma onto an LC-MS/MS instrument. With the optimal conditions, greater than 1,000 injections of diluted plasma have been successfully performed on a single column in less than 19 h making this technique an excellent approach for the rapid preparation and high throughput of plasma samples containing drug levels in the ng/ml range or higher. Application of this methodology to measure the levels of dextromethorphan in canine plasma to evaluate drug delivery from various formulations is also presented. PMID- 11022909 TI - Simultaneous spectrofluorimetric determination of (acetyl)salicylic acid, codeine and pyridoxine in pharmaceutical preparations using partial least-squares multivariate calibration. AB - A partial least-squares calibration (PLS) method for the simultaneous spectrofluorimetric determination of salicylic acid (SA), codeine (CO) and pyridoxine (PY) is proposed. The determination of SA, CO, and PY has been carried out in mixtures of up to three components by recording the emission fluorescence spectra between 300 and 500 nm (lambda(exc) = 220 nm). Due to the fact of the strong spectral overlap among the excitation and also among the emission spectra of these compounds, a previous separation should be carried out in order to determine them by conventional spectrofluorimetric methodologies. Here, a full spectrum multivariate calibration PLS method is developed. The experimental calibration matrix was constructed with 14 samples. The concentration ranges considered were 0.1-2.0 (SA), 0.25-3.0 (CO) and 0.10-2.0 (PY) mg x l(-1). The optimum number of factors was selected by using the cross-validation method. The method also allows the simultaneous determination of acetylsalicylic acid (ASA), CO and PY by previous alkaline hydrolysis of ASA to SA. To check the accuracy of the proposed method, it was applied to the determination of these compounds in synthetic mixtures and in pharmaceuticals. PMID- 11022910 TI - Facile and selective determination of the cerebral vasodilator nafronyl in a commercial formulation by heavy atom induced room temperature phosphorimetry. AB - This paper presents a facile and selective method for the determination of the pharmaceutical compound nafronyl using heavy atom induced room temperature phosphorimetry (HAI-RTP) as analytical technique. The determination was performed in potassium iodide 1.6 M and sodium sulphite 0.002 M at a measurement temperature of 20 degrees C. The phosphorescence intensity was then measured at lambda(exc) = 292 nm and lambda(em) = 524 nm. Phosphorescence was fully developed instantly, obtaining a linear concentration range between 2.7 and 250 ng ml(-1) with the detection limit of 2.7 ng ml(-1), an analytical sensitivity of 5.1 ng ml(-1) and a standard deviation of 2.17%, at a 150 ng ml (-1) concentration level. The proposed method has been satisfactorily applied to the unique Spanish commercial formulation containing nafronyl at a 100 mg level per capsule. The recovery was 108%, with a 1.7%, standard deviation of the analytical measurement. The method has been validated using standard addition methodology. PMID- 11022911 TI - Quantitative determination of pravastatin and its biotransformation products in human serum by turbo ion spray LC/MS/MS. AB - A sensitive, specific, accurate and reproducible analytical method was developed and validated to quantify pravastatin (Prav), pravastatin-d5 (Prav-d5), SQ-31906, SQ-31906-d5, and pravastatin lactone (Prav-Lac) in human serum samples. Serum samples (0.5 ml) were acidified and extracted by a solid-phase extraction procedure to isolate all five analytes from human serum. Sample extracts were reconstituted and analyzed by turbo ion spray liquid chromatography/tandem mass spectrometry (LC/MS/MS) in the positive ion mode. The total run time was 9 min between injections. The assay demonstrated a lower limit of quantitation (LLQ) of 0.5 ng/ml for all five analytes. The calibration curves were linear from 0.5 ng ml to 100 ng/ml for all five analytes. The coefficients of determination of all calibration curves were > or = 0.999. Precision and accuracy quality control (QC) samples were prepared at concentrations of 2, 30, 80, and 500 ng/ml for all analytes. The intra-assay and inter-assay precision calculated from QC samples were within 8%, for all analytes. The inter-assay accuracy calculated from QC samples was within 8% for all analytes. The extraction recoveries were > or = 90% for all analytes. Benchtop stability experiments in an ice-water bath ( < or = 10 degrees C) demonstrated that over time, Prav-Lac hydrolyzes to Prav in serum. Prav, Prav-d5, SQ-31906, and SQ-31906-d5 were stable under these conditions for up to 24 h. Hydrolysis was minimized by buffering the serum to pH 4.5 and maintaining the serum sample in an ice-water bath. All analytes were stable after three freeze/thaw cycles and in reconstitution solution after 1 week at 4 degrees C. Stability of all analytes in human serum was demonstrated after storage at -70 degrees C for 77 days. The benchtop (< or = 10 degrees C) stability of pooled study samples was also investigated and the results were comparable to those obtained from serum QC samples. PMID- 11022912 TI - A multi-element ICP-MS survey method as an alternative to the heavy metals limit test for pharmaceutical materials. AB - A multi-element inductively coupled plasma-mass spectrometry (ICP-MS) survey method has been demonstrated as an alternative to the antiquated 'heavy metals limit test' prescribed by United States Pharmacopoeia (USP), European Pharmacopoeia (EP), and British Pharmacopoeia (BP), for drug substances, intermediates, and raw materials. The survey method is simple, fast, sensitive, semi-quantitative to quantitative, and includes all the elements which can be analyzed by atomic spectroscopy. PMID- 11022913 TI - Analysis of tobramycin by liquid chromatography with pulsed electrochemical detection. AB - The determination of tobramycin by liquid chromatography using a column packed with poly(styrene-divinylbenzene) and pulsed electrochemical detection on a gold electrode is described. The mobile phase consisted of an aqueous solution containing 52 g/l of sodium sulfate, 1.5 g/l of sodium octanesulfonate and 50 ml/l of a 0.2 M phosphate buffer pH 3.0. The total time of analysis was not more than 30 min. The effects of the different chromatographic parameters on the separation were also investigated. When a number of commercial samples of tobramycin was analyzed using this method, nine different components were separated, five of which are of unknown identity. PMID- 11022914 TI - Direct determination of ecgonine methyl ester and cocaine in rat plasma, utilizing on-line sample extraction coupled with rapid chromatography/quadrupole orthogonal acceleration time-of-flight detection. AB - Our current experiments assess the applicability of on-line sample extraction with coupled rapid chromatography systems to quadrupole orthogonal acceleration time-of-flight (Q-TOF) detection for the quantitative analysis of cocaine (COC), and ecgonine methyl ester (EME) in rat plasma. Experiments were performed on a Q TOF instrument, operated in the MS/MS mode. Quantitation was achieved utilizing the most prominent parent-daughter transition and internal standard calibration techniques (COC-d3: IS). The calibration curves produced for EME and COC ranged from 5.0 to 10,000 and 0.5 to 10,000 ng/ml, respectively. Equations of regression line and correlation coefficients for the pseudo-multiple reaction monitoring (MRM) ion abundance ratio and the corresponding calibration concentrations (r2) were as follows: y = 0.0003 + 0.0703x (r2 = 0.9921) for EME and y = 0.0032 + 0.0035x (r2 = 0.9997) for COC. The system repeatability, given as percent coefficient of variation (% CV) of mean peak-area ratios, was assessed using 50 injections of a rat plasma sample from the pharmacokinetic study. The analyses were performed over the course of 5 days, rendering % CVs for EME and COC of 0.73 and 0.58, respectively. This method suggests that on-line sample extraction coupled with fast liquid chromatography/quadrupole orthogonal time-of-flight mass spectrometry may be a viable alternative for quantitative analysis of EME and COC in rat plasma. PMID- 11022915 TI - HPLC determination of guaifenesin with selected medications on underivatized silica with an aqueous-organic mobile phase. AB - A high performance liquid chromatography procedure has been developed for the simultaneous determination of guaifenesin pseudoephedrine-dextromethorphan and guaifenesin-pseudoephedrine in commercially available capsule dosage forms and guaifenesin-codeine in a commercial cough syrup dosage form. The separation and quantitation are achieved on a 25-cm underivatized silica column using a mobile phase of 60:40%) v/v 6.25 mM phosphate buffer, pH 3.0 - acetonitrile at a flow rate of 1 ml min(-1) with detection of all analytes at 216 nm. The separation is achieved within 10 min for each drug mixture. The method showed linearity for the guaifenesin-pseudoephedrine-dextromethorphan mixture in the 50-200, 7.5-30 and 2.5-10, microg ml(-1) ranges, respectively. The intra- and inter-day RSDs ranged from 0.23 to 4.20%, 0.18 to 2.85%, and 0.13 to 5.04% for guaifenesin, pseudoephedrine, and dextromethorphan, respectively. The guaifenesin pseudoephedrine mixture yielded linear ranges of 25-100 and 3.75-15 microg ml(-1) and intra- and inter-day RSDs ranged from 0.65 to 4.18% and 0.23 to 3.00% for guaifenesin and pseudoephedrine, respectively. The method showed linearity for the guaifenesin-codeine mixture in the 25-100 and 2.5-10 microg ml(-1) ranges and RSDs ranged from 0.37 to 4.25% and 0.14 to 2.08% for guaifenesin and codeine, respectively. PMID- 11022916 TI - Simultaneous analysis of naproxen, nabumetone and its major metabolite 6-methoxy 2-naphthylacetic acid in pharmaceuticals and human urine by high-performance liquid chromatography. AB - A high-performance liquid chromatographic (HPLC) method for simultaneous determination of naproxen (NAP), nabumetone (NAB) and its major metabolite, 6 methoxy-2-naphthylacetic acid (6-MNA), was developed for the application to pharmaceuticals and human urine. Isocratic reversed-phase HPLC was employed for quantitative analysis using triethylamine and 1-heptanesulfonic acid sodium salt (HSA) as ion-pair reagents. Urine samples were purified by solid-phase extraction using Bond-Elut Certify II cartridges containing reversed-phase and anion exchange functionalities. The HPLC assay was carried out using a Wakosil ODS 5C18 column (5 microm, 150 x 4.6 mm, i.d.). The mobile phase consisted of 0.5 g of HSA dissolved in 1,000 ml of a mixture of acetonitrile, water and triethylamine (500:500:1, v/v) adjusted with phosphoric acid to pH 3. The calibration curves of NAP and NAB showed good linearity in the concentration range 32-160 microg/ml with UV detection (270 nm) for pharmaceuticals. In the low concentration ranges (8-96 ng of NAP per ml, 24-288 ng of NAB per ml and 5.6-67.2 ng of 6-MNA per ml), the calibration curves were also obtained with fluorimetric detection (excitation 280 nm, emission 350 nm) for biological fluids. The correlation coefficients were better than 0.999 in all cases. The lower limits of detection (defined as a signal-to-noise ratio of about 3) were approximately 0.3 ng for NAP, 1.5 ng for NAB and 0.2 ng for 6-MNA. The procedure described here is rapid, simple, selective, and is suitable for routine analysis of pharmaceuticals and pharmacokinetic studies in human urine samples. PMID- 11022917 TI - Identification of autoxidation and photodegradation products of ethynylestradiol by on-line HPLC-NMR and HPLC-S. AB - HPLC-NMR, HPLC-MS, and HPLC-UV were used to characterize the predominant solution autoxidation and photodegradation products of ethynylestradiol (1). A hydroperoxide (2) and a series of isomeric dimeric oxidation products (3-7), were identified. PMID- 11022918 TI - Drug-company influence on medical education in USA. PMID- 11022919 TI - Health costs due to outdoor air pollution by traffic. PMID- 11022920 TI - Role of interferon-gamma gene in rheumatoid arthritis? PMID- 11022921 TI - Immunotherapy in hepatocellular carcinoma. PMID- 11022922 TI - Capsaicin and nociception: from basic mechanisms to novel drugs. PMID- 11022923 TI - Diarrhoea, diagnostic delay, and appendicitis. PMID- 11022924 TI - Reinvigorating public health. PMID- 11022925 TI - Neonatal screening for cystic fibrosis in Brittany, France: assessment of 10 years' experience and impact on prenatal diagnosis. AB - BACKGROUND: Neonatal screening for cystic fibrosis has been a subject of debate over the past few years. This study assesses 10 years of neonatal screening in Brittany, France, and examines its impact on prenatal screening of subsequent pregnancies in couples with an affected child. METHODS: The study included all the neonates screened for cystic fibrosis in Brittany from Jan 1, 1989, to Dec 31, 1998. The screening consisted of an immunoreactive trypsinogen assay from dried blood spots, plus, from 1993, mutation analysis. Data were collected on incidence of cystic fibrosis, and genotypic and biochemical characteristics. The use of prenatal screening of subsequent pregnancies in affected families was also investigated. FINDINGS: Of the 343,756 neonates screened, 118 children with cystic fibrosis were identified, giving an incidence of one in 2913. All mutated alleles were characterised: 34 different mutations resulting in 36 genotypes were detected. The introduction of DNA analysis into the protocol greatly reduced the recall rate and increased the sensitivity of the test. The mean cost of the screening programme was US$2.32 per screened child. 39 (34%) of the families identified by neonatal screening opted for subsequent prenatal diagnosis at least once. 12 couples would have benefited from this procedure while their first child was still symptom-free. 42 healthy children were born, and 18 pregnancies were terminated (therapeutic abortion rate of 100%). INTERPRETATION: We have shown the feasibility of neonatal screening for cystic fibrosis in Brittany. Through the detection of a large range of mutations, neonatal screening provides the opportunity for more reliable prenatal diagnosis and cascade screening. The neonatal screening programme described here could provide a good model for other countries intending to initiate such a scheme. PMID- 11022926 TI - Public-health impact of outdoor and traffic-related air pollution: a European assessment. AB - BACKGROUND: Air pollution contributes to mortality and morbidity. We estimated the impact of outdoor (total) and traffic-related air pollution on public health in Austria, France, and Switzerland. Attributable cases of morbidity and mortality were estimated. METHODS: Epidemiology-based exposure-response functions for a 10 microg/m3 increase in particulate matter (PM10) were used to quantify the effects of air pollution. Cases attributable to air pollution were estimated for mortality (adults > or = 30 years), respiratory and cardiovascular hospital admissions (all ages), incidence of chronic bronchitis (adults > or = 25 years), bronchitis episodes in children (< 15 years), restricted activity days (adults > or = 20 years), and asthma attacks in adults and children. Population exposure (PM10) was modelled for each km2. The traffic-related fraction was estimated based on PM10 emission inventories. FINDINGS: Air pollution caused 6% of total mortality or more than 40,000 attributable cases per year. About half of all mortality caused by air pollution was attributed to motorised traffic, accounting also for: more than 25,000 new cases of chronic bronchitis (adults); more than 290,000 episodes of bronchitis (children); more than 0.5 million asthma attacks; and more than 16 million person-days of restricted activities. INTERPRETATION: This assessment estimates the public-health impacts of current patterns of air pollution. Although individual health risks of air pollution are relatively small, the public-health consequences are considerable. Traffic-related air pollution remains a key target for public-health action in Europe. Our results, which have also been used for economic valuation, should guide decisions on the assessment of environmental health-policy options. PMID- 11022928 TI - Family support for stroke: a randomised controlled trial. AB - BACKGROUND: Attention is currently focused on family care of stroke survivors, but the effectiveness of support services is unclear. We did a single-blind, randomised, controlled trial to assess the impact of family support on stroke patients and their carers. METHODS: Patients with acute stroke admitted to hospitals in Oxford, UK, were assigned family support or normal care within 6 weeks of stroke. After 6 months, we assessed, for carers, knowledge about stroke, Frenchay activities index, general health questionnaire-28 scores, caregiver strain index, Dartmouth co-op charts, short form 36 (SF-36), and satisfaction scores, and, for patients, knowledge about stroke and use of services, Barthel index, Rivermead mobility index, Frenchay activities index, London handicap scale, hospital anxiety and depression scales, Dartmouth co-op charts, and satisfaction. FINDINGS: 323 patients and 267 carers were followed up. Carers in the intervention group had significantly better Frenchay activities indices (p=0.03), SF-36 scores (energy p=0.02, mental health p=0.004, pain p=0.03, physical function p=0.025, and general health perception p=0.02), quality of life on the Dartmouth co-op chart (p=0.01), and satisfaction with understanding of stroke (82 vs 71%, p=0.04) than those in the control group. Patients' knowledge about stroke, disability, handicap, quality of life, and satisfaction with services and understanding of stroke did not differ between groups. Fewer patients in the intervention group than in the control group saw a physiotherapist after discharge (44 vs 56%, p=0.04), but use of other services was similar. INTERPRETATION: Family support significantly increased social activities and improved quality of life for carers, with no significant effects on patients. PMID- 11022927 TI - Adoptive immunotherapy to lower postsurgical recurrence rates of hepatocellular carcinoma: a randomised trial. AB - BACKGROUND: Postsurgical recurrence of hepatocellular carcinoma (HCC) is frequent and fatal. Adoptive immunotherapy is active against HCC. We assessed whether postoperative immunotherapy could lower the frequency of recurrence. METHODS: Between 1992 and 1995, we did a randomised trial in which 150 patients who had undergone curative resection for HCC were assigned adoptive immunotherapy (n=76) or no adjuvant treatment (n=74). Autologous lymphocytes activated vitro with recombinant interleukin-2 and antibody to CD3 were infused five times during the first 6 months. Primary endpoints were time to first recurrence and recurrence free survival and analyses were by intention to treat. FINDINGS: 76 patients received 370 (97%) of 380 scheduled lymphocyte infusions (mean cell number per patient 7.1x10(10) [SD 2.1]; CD3 and HLA-DR cells 78% [16]), and none had grade 3 or 4 adverse events. After a median follow-up of 4.4 years (range 0.2-6.7), adoptive immunotherapy decreased the frequency of recurrence by 18% compared with controls (45 [59%] vs 57 [77%]) [corrected] patients. Time to first recurrence in the immunotherapy group was significantly longer than that in the control group (48% [37-59] vs 33% [22-43] at 3 years, 38% [22-54] vs 22% [11-34] at 5 years; p=0.008). The immunotherapy group had significantly longer recurrence-free survival (p=0.01) and disease-specific survival (p=0.04) than the control group. Overall survival did not differ significantly between groups (p=0.09). INTERPRETATION: Adoptive immunotherapy is a safe, feasible treatment that can lower recurrence and improve recurrence-free outcomes after surgery for HCC. PMID- 11022929 TI - Intergenerational continuity of child physical abuse: how good is the evidence? AB - BACKGROUND: There is widespread belief that individuals who were physically abused during childhood are more likely to abuse their own children than those who were not abused, but the empirical studies examining this belief have not been systematically reviewed. The aim of this study was to evaluate systematically, based on eight methodological standards derived from a hypothetical randomised controlled trial, the design of studies investigating the intergenerational transmission of child physical abuse. METHODS: We reviewed studies published between 1965 and 2000 in English that provided information about physical maltreatment in two generations and included a comparison group. Two investigators independently assessed whether each study met the methodological standards. FINDINGS: In the ten studies identified (four cohort, one cross-sectional, and five case-control), the relative risks of maltreatment in the children of parents who were abused during childhood were significantly increased in four studies (relative risks 4.75-37.8), but in three other studies the relative risks were less than 2. Most study reports provided a clear description of abuse of parents during childhood and abuse of their children. Five studies failed to avoid recall and detection bias; five did not ensure that controls were not themselves maltreated; eight did not provide adequate follow up; and in six the report did not state whether the enrolled parent was responsible for the maltreatment. Most studies did not control for intervening factors, such as sociodemographic characteristics during the time of abuse of the parent generation and at the time their children were abused. Only one study met all eight criteria (relative risk of abuse transmission 12.6 [95% CI 1.82-87.2]) and one met six (1.05 [0.53-2.06]). INTERPRETATION: The one study that met all eight methodological standards provided evidence for the intergenerational continuity of child physical abuse, but that which met six standards did not support the hypothesis. Use of our model and methodological standards should improve the scientific quality of studies examining the effects of risk factors for adverse outcomes that may continue across generations. PMID- 11022930 TI - Association between dinucleotide repeat in non-coding region of interferon-gamma gene and susceptibility to, and severity of, rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis ranges from a mild, non-deforming arthropathy with little long-term disability to severe, incapacitating, deforming arthritis which may be refractory to conventional disease-modifying agents. Epidemiological studies show an important genetic influence in rheumatoid arthritis, and MHC region genes and cytokine genes within and outside this region have been considered as candidates. We did a case-control study to test whether polymorphisms in the interferon-gamma gene are associated with severity of rheumatoid arthritis. METHODS: Interferon gamma dinucleotide repeat polymorphisms were examined with quantitative genescan technology, and HLA-DR alleles were identified by PCR and restriction-fragment-length polymorphism analysis. We studied 60 patients with severe rheumatoid arthritis, 39 with mild disease, and 65 normal controls. FINDINGS: Susceptibility to, and severity of, rheumatoid arthritis were related to a microsatellite polymorphism within the first intron of the interferon-gamma gene. A 126 bp allele was seen in 44 (73%) of 60 patients with severe rheumatoid arthritis, compared with eight (21%) of 39 with mild disease (odds ratio 10.66 [95% CI 4.1-24.9]), and with eight (12%) of 65 normal controls (19.59 [7.7-49.9]). Conversely, a 122 bp allele at the same locus was found in four (7%) patients with severe disease compared with 25 (64%) of those with mild disease (0.04 [0.01-0.1]) and with 52 (80%) of controls (0.018 [0.005 0.06]). INTERPRETATION: This association may be valuable for understanding the mechanism of disease progression, for predicting the course of the disease, and for guiding therapy. PMID- 11022931 TI - Bulbar palsies and chest pain. PMID- 11022932 TI - Human brain-cell death induced by tumour-necrosis-factor-related apoptosis inducing ligand (TRAIL) AB - Cell death induced by tumour-necrosis-factor-related apoptosis-inducing ligand (TRAIL) was believed to occur exclusively in tumour cells, suggesting that this drug is safe to use as an antitumour therapy. Concerns were raised, however, when cultured normal human hepatocytes were shown to be susceptible to TRAIL. Here we report that TRAIL induces apoptosis in the human brain. Our finding therefore argues against the use of TRAIL for therapy of human brain tumours. However, neuroinflammatory T cells that express TRAIL might induce apoptosis of brain tissue, indicating a potential target for treatment of multiple sclerosis. PMID- 11022933 TI - Antibacterial properties of xanthine oxidase in human milk. AB - Formula-fed babies contract gastroenteritis more than breast-fed babies, which is of concern to mothers who cannot breastfeed or, as with HIV-infected mothers, are discouraged from breastfeeding. The ability of endogenous breastmilk xanthine oxidase to generate the antimicrobial radical nitric oxide has been measured and its influence on the growth of Escherichia coli and Salmonella enteritides examined. Breastmilk, but not formula feed, generated nitric oxide. Xanthine oxidase activity substantially inhibited the growth of both bacteria. An important natural antibiotic system is missing in formula feeds; the addition of xanthine oxidase may improve formula for use when breastfeeding is not a safe option. PMID- 11022934 TI - Somatic mutation in MECP2 as a non-fatal neurodevelopmental disorder in males. AB - Rett syndrome is a cause of severe learning disability in girls and is associated with a characteristic history and movement disorder. It is an X-linked dominant condition associated with mutations of the MECP2 gene on the distal part of the X chromosome. If present in a male conceptus, the mutation is usually lethal. We present evidence to show that males can be affected by Rett syndrome. In the boy presented, this situation came about because cells containing the MECP2 mutation existed alongside a normal cell line. Somatic mosaicism could explain the occurrence of other X-linked dominant disorders in males, when they would normally be lethal. PMID- 11022935 TI - Endovascular stenting of abdominal aortic aneurysm in patients unfit for elective open surgery. Eurostar group. EUROpean collaborators registry on Stent-graft Techniques for abdominal aortic Aneurysm Repair. AB - Endovascular aneurysm repair is useful for patients who are judged unfit for surgery. We investigated the outcome of endovascular repair of abdominal aortic aneurysm in patients fit and unfit for surgery. The 1-year cumulative survival for patients unfit for surgery and patients unfit for general anaesthesia was 20% and 23%, respectively. The overall health status of patients was an important predictor of survival after endovascular repair. The risks of endovascular aneurysm repair might, therefore, exceed that of non-operative management. Caution should be used when advising these patients about endovascular repair. PMID- 11022936 TI - Dominic O'Brien--master mnemonist. PMID- 11022937 TI - Gore scores in voter polls with health-care proposals. PMID- 11022938 TI - Conflict of interest taints vaccine approval process, charges US report. PMID- 11022940 TI - UN report highlights health effects of sanctions on Afghanistan. PMID- 11022939 TI - Uterine transplantation failure causes Saudi Arabian government clampdown. PMID- 11022941 TI - WHO to support health reforms in South East Asian region. PMID- 11022942 TI - HIV murders prompt activist anger. PMID- 11022943 TI - Public health in developing countries. AB - Poverty not only excludes people from the benefits of health-care systems but also restricts them from participating in decisions that affect their health. The resulting health inequalities are well documented, and the search for greater equity attracts many concerned players and initiatives. Fundamental to the success of these efforts, however, is the need for people to be able to negotiate their own inclusion into health systems and demand adequate health care. This calls for a restatement of the centrality of people in public health and its practice. New forms of communication and cooperation are required at all levels of society, nationally, and internationally, to ensure equitable exchange of views and knowledge to formulate appropriate action to redress inequalities and improve people's health and wellbeing. PMID- 11022944 TI - A case of twin-to-twin transfusion in 1617. PMID- 11022945 TI - Patents and intellectual property: a salvation for patient-oriented research? PMID- 11022946 TI - Comparison of losartan and captopril in ELITE II. PMID- 11022947 TI - Comparison of losartan and captopril in ELITE II. PMID- 11022948 TI - Comparison of losartan and captopril in ELITE II. PMID- 11022949 TI - Comparison of losartan and captopril in ELITE II. PMID- 11022950 TI - Regular use of salbutamol in asthma. PMID- 11022951 TI - Treatment for patients with stage-1 endometrial carcinoma. PMID- 11022952 TI - Premature cardiovascular disease and chronic renal failure. PMID- 11022953 TI - Carbaril for head lice. PMID- 11022954 TI - Birth size and arterial compliance in young adults. PMID- 11022955 TI - Mono-ocular occlusion for treatment of dyslexia. PMID- 11022956 TI - The poor man cooks with water. PMID- 11022957 TI - Tumour site, sex, and survival in colorectal cancer. PMID- 11022958 TI - Tumour site, sex, and survival in colorectal cancer. PMID- 11022959 TI - Tumour site, sex, and survival in colorectal cancer. PMID- 11022960 TI - Doctors in the NHS. National Health Service. PMID- 11022961 TI - Doctors in the NHS. National Health Service. PMID- 11022962 TI - Doctors in the NHS. National Health Service. PMID- 11022963 TI - Doctors in the NHS. National Health Service. PMID- 11022964 TI - Severe malaria with impaired cerebral function? PMID- 11022965 TI - Boycott of conferences in Austria. PMID- 11022966 TI - Familial and acquired long qt syndrome and the cardiac rapid delayed rectifier potassium current. AB - 1. Long QT syndrome (LQTS) is a cardiac disorder characterized by syncope, seizures and sudden death; it can be congenital, idiopathic, or iatrogenic. 2. Long QT syndrome is so-named because of the connection observed between the distinctive polymorphic ventricular tachycardia torsade de pointes and prolongation of the QT interval of the electrocardiogram, reflecting abnormally slowed ventricular action potential (AP) repolarization. Acquired LQTS has many similar clinical features to congenital LQTS, but typically affects older individuals and is often associated with specific pharmacological agents. 3. A growing number of drugs associated with QT prolongation and its concomitant risks of arrhythmia and sudden death have been shown to block the 'rapid' cardiac delayed rectifier potassium current (IKr) or cloned channels encoded by the human ether-a-go-go-related gene (HERG; the gene believed to encode native IKr). Because IKr plays an important role in ventricular AP repolarization, its inhibition would be expected to result in prolongation of both the AP and QT interval of the electrocardiogram. 4. The drugs that produce acquired LQTS are structurally heterogeneous, including anti-arrhythmics, such as quinidine, non sedating antihistamines, such as terfenadine, and psychiatric drugs, such as haloperidol. In addition to heterogeneity in their structure, the electrophysiological characteristics of HERG/IKr inhibition differ between agents. 5. Here, clinical observations are associated with cellular data to correlate acquired LQTS with the IKr/HERG potassium (K+) channel. One strategy for developing improved compounds in those drug classes that are currently associated with LQTS could be to design drug structures that preserve clinical efficacy but are modified to avoid pharmacological interactions with IKr. Until such time, awareness of the QT-prolongation risk of particular agents is important for the clinician. PMID- 11022967 TI - Effects of the new hydroxy-3-methylglutaryl coenzyme a reductase inhibitor fluvastatin on anti-oxidant enzyme activities and renal function in streptozotocin-induced diabetic rats. AB - 1. The effects of 11 week treatments with the new hydroxy3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor fluvastatin on renal intrinsic anti oxidant enzyme (AOE) activities and renal function were evaluated in streptozotocin (STZ)-induced diabetic rats. 2. Renal intrinsic AOE activities, creatinine clearance and urinary albumin excretion were examined in STZ-induced diabetic rats. The levels of total cholesterol (TC), triglyceride (TG) and phospholipid (PL) were also examined. 3. In general, renal AOE activities and function were lower in diabetic rats than in non-diabetic Sprague-Dawley rats. 4. Decreases in TC, TG and PL levels and urinary albumin excretion by the HMG-CoA reductase inhibitor fluvastatin improved renal function and produced a non uniform alteration in renal AOE; only glutathione peroxidase (GSH-Px) activity increased significantly with fluvastatin treatment. 5. It appears that the improvement in renal function and albuminuria may be related to increases in GSH Px activity, but there was no correlation between changes in renal function and changes in the activity of Mn-superoxide dismutase or catalase. PMID- 11022968 TI - Mechanisms of alpha,beta-methylene atp-induced inhibition in rat ileal smooth muscle: involvement of intracellular Ca2+ stores in purinergic inhibition. AB - 1. In order to investigate purinergic effects on rat ileal smooth muscle, we used alpha,beta-methylene ATP (alpha,beta-MeATP), ATP, ADP and UTP. alpha,beta Methylene ATP and ATP were the only agonists that caused a concentration dependent inhibition of carbachol-precontracted smooth muscle. The inhibitory effect of alpha,beta-MeATP was completely blocked by pyridoxalphosphate-6 azophenyl-2',4'-disulphonic acid (3 x 10(-5) mol/L), a selective antagonist of the P2X > > P2Y receptor. 2. Using reverse transcription-polymerase chain reaction we demonstrated the presence of both, P2X and P2Y receptor mRNA within the rat ileal longitudinal muscle/myenteric plexus layer preparation. 3. The alpha,beta-MeATP-induced inhibition was blocked in a concentration-dependent manner in the presence of the K+ channel blocker apamin, but was unaffected by other K+ channel blockers, such as charybdotoxin (10(-7) mol/L), 4-aminopyridine (10(-4)mol/L), glibenclamide (10(-5) mol/L) and tetraethylammonium (10(-3) mol/L). 4. The alpha,beta-MeATP-induced inhibition was unaffected by pretreatment with atropine (10(-6) mol/L), phentolamine (10(-6) mol/L), propranolol (10(-6) mol/L), nitrendipine (10(-7) mol/L), pertussis toxin (10(-6) mol/L) NG-nitro-L arginine (3 x 10(-4) mol/L) and tetrodotoxin (10(-6) mol/L), excluding an involvement of adrenergic, cholinergic, neural, nitrinergic or G-protein involvement in purinergic-mediated inhibition. 5. In order to investigate whether the internal Ca2+ stores participated in the inhibitory effect observed, we depleted internal Ca2+ stores with cyclopiazonic acid, a specific Ca2+-ATPase inhibitor. The inhibitory effect of alpha,beta-MeATP was completely abolished after depletion of the intracellular Ca2+ stores. 6. This is in contrast with the effects seen for neurotensin, where neurotensin-induced inhibition was unchanged after depletion of intracellular Ca2+ stores, suggesting at least two different pathways of apamin-sensitive non-adrenergic, non-cholinergic inhibition in rat ileal smooth muscle. 7. According to our results, the inhibitory effect of alpha,beta-MeATP in rat ileum longitudinal smooth muscle is mediated via a P2 purinoceptor, most likely a P2X receptor, involves G-protein-independent activation of an apamin-sensitive K+ channel and requires filled intracellular Ca+ stores. PMID- 11022969 TI - Garlic elicits a nitric oxide-dependent relaxation and inhibits hypoxic pulmonary vasoconstriction in rats. AB - 1. The aims of the present study were to determine the characteristics of garlic extract-induced relaxation in rat isolated pulmonary arteries, its susceptibility to changes in oxygen tension and its protective effect against hypoxic pulmonary vasoconstriction. 2. In normoxia, garlic extract (3-500 microg/mL) produced a dose- and nitric oxide (NO)-dependent relaxation. Following 60 min hypoxia, maximum garlic relaxation was reduced compared with control (mean (-SEM) -86 +/- 3 vs-69 +/- 2% of phenylephrine (PE) precontraction, respectively), but recovered after 60 min reoxygenation (-85 +/- 3% PE precontraction). 3. Acetylcholine (0.1 micromol/L)-induced NO-dependent relaxation was reduced from a control value of 76 +/- 1% to -46 +/- 4% during hypoxia and was further reduced to -35 +/- 2 % after reoxygenation. 4. In endothelium-intact arteries, hypoxic exposure resulted in a triphasic response: early transient contraction (+24 +/- 4%), followed by transient relaxation (-37 +/- 7%) and then sustained contraction (+62 +/- 5%). 5. Pretreatment with NG-nitro-L-arginine methyl ester abolished the early transient contraction, moderately attenuated the sustained contraction and had no effect on the transient relaxation. Mechanical endothelial disruption inhibited all hypoxia induced vascular changes. 6. Garlic pretreatment had no effect on the early transient contraction (+25 +/- 4%), but inhibited the transient relaxation (-5 +/ 3%; P<0.05) and the sustained contraction (+26 +/- 5%; 7. Garlic also significantly inhibited endothelin-l-induced contractions in a dose-dependent manner. 8. These findings show that garlic extract modulates the production and function of both endothelium-derived relaxing and constricting factors and this may contribute to its protective effect against hypoxic pulmonary vasoconstriction. PMID- 11022970 TI - Inhibition of cytokine release by and cardiac effects of type IV phosphodiesterase inhibition in early, profound endotoxaemia in vivo. AB - 1. In rats, inhibition of type IV phosphodiesterase (PDE4) attenuates acute renal failure and early (hours) mortality induced by high-dose endotoxin. Because it is unlikely that protection of renal function accounts for improved early survivability, most likely PDE4 inhibition exerts multiple beneficial effects in endotoxaemia and the purpose of the present study was to test this hypothesis. 2. In study 1, we determined, in anaesthetized rats, the effects of endotoxin (30 mg/kg, i.v.) on cardiac performance parameters (heart rate (HR), ventricular peak systolic pressure (VPSP), maximum positive change in left ventricular pressure with respect to time (+dP/dt), maximum negative change in left ventricular pressure with respect to time (-dP/dtmax), ventricular end-diastolic pressure (VEDP), ventricular minimum diastolic pressure (VMDP) and HR-pressure product), plasma catecholamine levels, plasma renin activity (PRA) and plasma levels of inflammatory cytokines (tumour necrosis factor (TNF)-alpha and interleukin (IL) lbeta). 3. In study 2, we determined, in anaesthetized rats, whether inhibition of PDE4 attenuates lipopolysaccharide (LPS)-induced changes in the aforementioned parameters of heart performance and neurohumoral status. We compared the changes in these parameters induced by endotoxaemia in animals treated with either RO 20 1724 (10 microg/kg per min; a selective PDE4 inhibitor) or its vehicle (DMSO; 1.35 microL/min). 4. At 90 min postadministration, endotoxin significantly increased HR and reduced -dP/dtmax and VEDP and caused a several-fold increase in plasma levels of TNF-alpha, IL-1beta, noradrenaline, adrenaline and PRA. RO20 1724 significantly blunted the endotoxin-induced reduction in -dP/dtmax and decreased endotoxin-induced increases in TNF-alpha and IL-1beta without significantly altering endotoxin-induced changes in HR, VEDP, catecholamine levels and PRA. 5. Results from these studies indicate that, in addition to preserving renal function, PDE4 inhibition attenuates inflammatory cytokine release caused by high-dose endotoxin and may have protective effects on diastolic function in early profound endotoxaemia. PMID- 11022971 TI - Metabolic variability among disease-resistant inbred rat strains and in comparison with wild rats (Rattus norvegicus). AB - 1. Inbreeding and optimization of environmental conditions for laboratory rats may have led to the survival of mutants with metabolic aberrations but without evident disease phenotype. Therefore, in the present study, we compared metabolic traits between so-called disease-resistant inbred rat strains Dark Agouti (DA), Brown Norway (BN), Lewis (LEW), Wistar-Kyoto (WKY), Fischer 344 (F344) and wild rats (Rattus norvegicus). 2. Twelve males of each strain at 12, 13 and 14 weeks of age were studied for bodyweight, body mass index (BMI), blood glucose, serum triglycerides, total cholesterol, insulin and leptin. 3. In comparison with wild rats, the cholesterol values were significantly increased in all inbred rats studied. Except for DA rats, all rats were also significantly heavier than wild rats. 4. There were also significant differences between the different disease resistant strains and WKY rats were the biggest animals with the highest bodyweight, BMI and cholesterol values. 5. The strains could be separated into groups with either very high (F344, LEW, WKY) or low values (DA, BN) of serum insulin and leptin levels. 6. Because all rats were studied under the same conditions, the findings suggest a substantial strain dependence in feeding behaviour and energy balance caused by the different genotypes. PMID- 11022972 TI - Relevance of cytochrome P450 levels in the actions of enflurane and isoflurane in mice: studies on the haem pathway. AB - 1. The effect of the fluorinated ether anaesthetics enflurane and isoflurane in mice on haem metabolism and regulation in different metabolic states, such as depression and induction of cytochrome P450 produced by allylisopropylacetamide (AIA) and imidazole, respectively, was investigated. 2. Mice previously treated with AIA (350 mg/kg, i.p.) or imidazole (400 mg/kg, i.p.) received a single dose (1 mL/kg, i.p.) of enflurane or isoflurane and were killed 20 min after anaesthetic administration. 3. Induction of delta-aminolevulinic acid synthetase (ALA-S) activity was found, as expected, in animals receiving AIA and also in animals treated with AIA plus anaesthesia, but no change in the activity of either porphobilinogenase (PBGase) or porphobilinogen deaminase (PBG-D) activities was detected in these two groups of animals. An additional increase in haem destruction was observed in the AIA plus isoflurane-treated group. When mice were injected with imidazol alone or in combination with the anaesthetics, ALA-S activity was increased 50-90% in all groups, but again no change in PBGase or PBG D activity was observed. Haem oxygenase was diminished in mice receiving imidazole and anaesthesia. 4. In conclusion, neither enflurane nor isoflurane caused additional disturbances in haem metabolism to those produced by AIA or imidazole alone. PMID- 11022973 TI - Dexmedetomidine and haemodynamic responses to acute central hypovolaemia in conscious rabbits. AB - 1. Effects of the alpha2-adrenoceptor agonist dexmedetomidine on vasoconstrictor and heart rate (HR) responses to acute central hypovolaemia were studied in eight chronically instrumented rabbits. We compared intravenous (i.v.) and fourth ventricular (V4) dexmedetomidine (0.1-10 microg/kg) and the reversal of effects by the alpha2-adrenoceptor antagonist idazoxan and the opioid agonist alfentanil. 2. Gradual inflation of an inferior vena cava (IVC) cuff reduced cardiac index (CI) by 8%/min, with progressive vasoconstriction and increased HR. In control rabbits, at approximately 40% baseline CI, there was sudden decompensation with failure of vasoconstriction and a fall in mean arterial pressure (MAP). 3. Dexmedetomidine (i.v. and V4) reduced resting MAP and HR and caused an earlier decompensation during central hypovolaemia. Intravenous dexmedetomidine (3 and 10 microg/kg) also reduced the slope of the initial vasoconstrictor response and the maximum HR. 4. The effects of dexmedetomidine were reversed by the antagonist idazoxan, which prevented the decompensation phase. Intravenous alfentanil was also effective in restoring the vasoconstrictor response and delaying decompensation with hypovolaemia after dexmedetomidine. Combining dexmedetomidine with an opioid, such as alfentanil, may provide the benefit of reduced sympathetic tone without increased risk of cardiovascular collapse. PMID- 11022974 TI - Cardioprotective effects of nicorandil in rabbits anaesthetized with halothane: potentiation of ischaemic preconditioning via KATP channels. AB - 1. The roles of ATP-sensitive K+ channels (KATP channels) in ischaemic or pharmacological preconditioning in the rabbit heart remain unclear. Infarct limitation by ischaemic preconditioning was abolished by the KATP channel blocker glibenclamide under ketamine/xylazine anaesthesia, but not under anaesthesia induced by pentobarbital. Infarct limitation by the KATP channel opener pinacidil was detected under ketamine/xylazine anaesthesia, but not under pentobarbital anaesthesia. Thus, these effects appear to be anaesthetic dependent. 2. In the present study, we examined whether nicorandil (a KATP channel opener nitrate) exhibits cardioprotective actions under halothane anaesthesia, another commonly used volatile anaesthetic. Control animals were subjected to 40 min coronary occlusion and 120 min reperfusion. Before 40 min ischaemia, the nicorandil group received nicorandil (100 microg/kg per min, i.v., for 10 min), the 5' preconditioning (PC) group received 5 min ischaemia/20 min reperfusion, the 2.5'PC group received 2.5 min preconditioning ischaemia/20 min reperfusion, the nicorandil +2.5'PC group received both nicorandil and 2.5 min ischaemia/20 min reperfusion, the nicorandil +2.5'PC + 5-hydroxydecanoate (5HD) group received both nicorandil and 2.5 min ischaemia/20 min reperfusion in the presence of 5 hydroxydecanoate (5HD; a KATP blocker) and the 5HD group received 5 mg/kg, i.v., 5HD alone. Myocardial infarct size in control (n = 7), nicorandil (n = 5), 5'PC (n = 8), 2.5'PC (n = 5), nicorandil + 2.5'PC (n = 5), nicorandil + 2.5'PC + 5HD (n = 5) and 5HD (n = 4) groups averaged 44.4 +/- 3.6, 41.7 +/- 5.7, 17.8 +/- 3.2,* 34.1 +/- 4.8, 21.3 +/- 4.2,* 39.1 +/- 5.6 and 38.9 +/- 5.0% of the area at risk, respectively (*P <0.05 vs control). 3. Thus, nicorandil alone did not have an infarct size-limiting effect in halothane-anaesthetized rabbits. However, the results suggest that even when nicorandil alone does not demonstrate a direct cardioprotective effect, it may enhance ischaemic preconditioning via KATP channels. Key words: ATP-sensitive K+ (KATP) channel, ischaemic preconditioning, myocardial infarction, nicorandil, rabbit. PMID- 11022976 TI - Introduction: a brief history of capillaries and some examples of their apparently strange behaviour. AB - 1. Over the past decade, evidence has accumulated challenging the notion that the smallest blood vessels, the capillaries, are static, uniform, semipermeable pipes with no function except as a passive barrier marking the boundary between the vascular compartments from the tissue compartment. 2. Instead, capillaries are dynamic structures that participate in the active regulation of water, waste and nutrient exchange, the formation and destruction of exchange vessels and the sites for initiation of signals to regulate the flow of blood into the exchange vascular network. 3. The following papers will provide insight into the expanded appreciation of the varied nature of capillaries and how their functions are co ordinated to achieve 'whole organ' exchange. PMID- 11022975 TI - Glomerular expression of endothelial nitric oxide synthase in deoxycorticosterone acetate-salt-treated rats. AB - 1. In the present study, we investigated the relationship between chronic volume loading and glomerular endothelial nitric oxide synthase (eNOS) expression. Immunohistochemical expression of eNOS in glomeruli was semiquantified by graded scores of staining intensity. Each glomerulus was isolated by microdissection and mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) in desoxycorticosterone acetate (DOCA)-salt-treated rats. 2. Glomerular expansion and dilatation of tubules were the main histological findings and glomerular and vascular injury scores were significantly higher in the DOCA-salt treated group than in the control group. Endothelial NOS staining in glomeruli in DOCA-salt-treated animals was 81.4% lower than in control animals. Endothelial NOS mRNA was also detected at a very low rate in the kidney of treated rats compared with control rats (22/80 vs 74/80 glomeruli, respectively). 3. These results suggest that eNOS protein and mRNA expression in glomeruli was reduced by DOCA-salt loading. Chronic volume loading may damage the glomerulus and this may be mediated, at least in part, by disruption of eNOS. PMID- 11022977 TI - Role for capillaries in coupling blood flow with metabolism. AB - 1. Skeletal muscle blood flow is coupled with metabolism; for this coupling to be effective in matching blood flow to capillary exchange, control of capillary blood flow and recruitment must reside at the capillary level. 2. Capillaries are, indeed, capable of sensing and responding to vasoactive stimuli. We report studies that indicate that contraction of skeletal muscle fibres underneath capillaries is capable of increasing blood flow in those capillaries. 3. This presumed metabolically related signal initiates remote dilations in arterioles upstream of the stimulated capillaries. Our findings indicate that the vasodilatory signal is transmitted along the blood vessel wall. 4. Although we present evidence supporting a role for gap junctionally mediated communication of this vasodilatory signal, it appears unlikely to be primarily electrotonic spread of membrane potential changes. 5. Our studies further indicate that the transmitted signal is not dependent on changes in endothelial cell calcium. PMID- 11022978 TI - Capillary network patterning during angiogenesis. AB - 1. Although much is known of signalling events associated with angiogenesis, starting with a sprout, less is known of how an intact microvascular network is correctly formed following the initiation of sprouting. The aim of the present report is to evaluate some of the factors that could be involved in directing the patterning of the resulting microvascular bed. 2. Hypothetical 'patterning rules' are discussed and considered in the context of data obtained from studies of angiogenesis. The guidelines suggest that both tissue- and stimulus-specific regulators are involved in directing patterning. 3. Examples of 'patterned' and 'non-patterned' angiogenesis in rat mesentery are given, with evidence to support stimulus-related differences in angiogenesis patterns. 4. Skeletal muscle angiogenesis follows metabolic patterns within the muscle, as discerned through traditional histochemical evaluation. However, further examination, using confocal microscopy and new patterning assays, indicates that branching patterns differ according to stimulus. Patterning guidelines within the tissue may be important in preventing the growth of sinusoidal vessels. 5. The concept of 'angiotypes' is proposed to direct future studies towards understanding how therapeutic angiogenesis or angiostasis applications can be used to obtain correctly patterned microvascular networks. Angiogenesis via 'branching' compared with 'non-branching' angiotypes will involve different signalling pathways. Both sprouting and dividing of capillaries are found in the 'branching' angiotype, with each pattern having several subtypes that would be expected to be mediated via different signalling mechanisms. PMID- 11022979 TI - Microvascular remodelling in chronic airway inflammation in mice. AB - 1. Chronic inflammation is associated with blood vessel remodelling, including vessel proliferation and enlargement, and changes in vessel phenotype. We sought to characterize these changes in chronic airway inflammation and to determine whether corticosteroids that inhibit inflammation, such as dexamethasone, can also reduce microvascular remodelling. 2. Chronic airway inflammation was induced in C3H mice by infection with Mycoplasmapulmonis and the tracheal vessels treatment also decreased the immunoreactivity for P-selectin and the number of adherent leucocytes (595 +/- 203 vs 2,024 +/- 393 cells/ mm2 in treated and non treated infected mice, respectively). 6. We conclude that microvascular enlargement and changes in vessel phenotype are features of some types of chronic inflammation and, furthermore, that dexamethasone reverses the microvascular enlargement, changes in vessel phenotype and leucocyte influx associated with chronic inflammatory airway disease. PMID- 11022980 TI - Diversity within pericytes. AB - 1. Pericytes are cells of microvessels (arterioles, capillaries and venules) that wrap around endothelial cells. They are most abundant on venules and are common on capillaries. 2. The pericyte population is highly variable between different tissues and organs, probably in a manner reflecting postarteriolar hydrostatic pressures. Pericytes are more abundant in the distal legs and feet, again suggesting a hydrostatic pressure-driven mechanical role for pericytes as protectors of microvessel wall integrity. 3. Pericyte alteration or degeneration is linked directly with microangiopathy in diabetes, scleroderma, hypertension, dementias and, possibly, inappropriate calcification of blood vessels. 4. Pericytes are functionally codependent on endothelial cells. Each cell type influences each others' mitotic rate and probably phenotypic expression. 5. Pericytes are not randomly located around microvessels. Instead, they are located adjacent to or over endothelial cell junctions of venules and especially over gaps between endothelial cells during inflammation. Pericytes are emerging as essential components of the microvessel wall, with metabolic, signalling and mechanical roles to support the endothelial cell. PMID- 11022981 TI - The microvasculature as a dynamic regulator of volume and solute exchange. AB - 1. It has long been held that the microvessels are passive pipes through which the formed elements of blood and plasma course and from which exchange occurs. Physiological and structural data from vascular cells in culture to whole organs strongly support the notion that microvessels, whether in their arrangement as networks, in their microvascular phenotypic subtypes or with regard to their permeability properties, are anything but static. 2. Examples from different models, organs and species are presented to illustrate the spatial and dynamic nature of the microvasculature. 3. Given that the ultimate goal of this cardiovascular research is to apply what is learned from component parts of the microvasculature to understand integrated tissue function, examples of the danger of generalizing results from limited studies are presented. 4. Finally, accurate interpretation of most of the research in this area is predicated on the assumption that probe molecules, rendered visible by the binding of dyes, are biologically inert. 5. We present data demonstrating that some dyes used widely in microvascular studies can alter the barrier under study and/or alter the physicochemical characteristics of the molecule to which they were bound. PMID- 11022983 TI - Intravenous magnesium sulfate in acute severe asthma. AB - OBJECTIVE: Intravenous magnesium sulfate (MgSO4), as an adjunctive medication to the standard treatment of acute asthma, improves admission rate or severity score in acute severe asthma patients. METHODOLOGY: We conducted a randomized double blind placebo controlled trial with subjects from the emergency room, Ramathibodi Hospital, Bangkok, Thailand. Patients, aged 15-65 years with acute severe asthma attack, whose severity scores were greater than 4 and who were willing to be enrolled in a study during March to November 1997 participated in the study. Randomly allocated patients received either 2 g intravenous MgSO4 or placebo, sterile water, as an adjunctive medication to standard therapy for acute asthma. The medication was diluted in 50 mL of 0.9% normal saline. MEASUREMENT: Severity scores were measured by two investigators using Fischl's indices. The times interval of measurements were at the initial (0), 60, 120, 180, and 240 min from receipt of treatment. Patients were hospitalized if the severity scores at 240 min exceeded 1. Risk ratio (RR) and 95% confidence interval (CI) of RR were applied to estimate the risk of admission. Analysis of variance with repeated measurement on time was used to determine the severity score between two groups. RESULTS: Thirty-four patients with acute severe asthma were enrolled in the present study. One patient from the placebo group was excluded because he did not consent to undergoing peak expiratory flow rate. Seventeen patients received MgSO4 and 16 patients received placebo. The general characteristics between the two groups were not significantly different, which reflected the quality of randomization. The admission rates of the placebo and MgSO4 group were 25.00% and 17.65%, respectively. Patients who received MgSO4 had preventive risk to be hospitalized 0.71 times relative to patients who received placebo. However, this preventive risk did not reach statistical significance (95% CI of RR = 0.19 2.67). The severity score at any time between the two groups was also not statistically significantly different (P = 0.366). CONCLUSION: With the present evidence, the hypothesis was not confirmed. Magnesium sulphate as an adjunct to standard therapy did not improve either admission rate or severity score in patients with acute severe asthma. PMID- 11022982 TI - Composition of alveolar surfactant changes with training in humans. AB - OBJECTIVE: We test the hypothesis that the changes we observed previously in the relative amounts of disaturated phospholipids (DSP), cholesterol (CHOL), and surfactant protein-A (SP-A) in human alveolar surfactant in response to acute exercise, and which were related to fitness, can be induced by training. METHODOLOGY: We examine the effect of 7 weeks' training on these major surfactant components, together with surfactant protein-B (SP-B), in bronchoalveolar lavage fluid harvested from 17 males, both at rest and after acute exercise. Fitness was assessed as workload/heart rate achieved during cycling for 30 min at 90% of theoretical maximal heart rate, and was increased in all subjects following training (mean increase 22.2+/-3.91%; P = 0.001). RESULTS: Training significantly increased the SP-A/DSP, SP-B/DSP, SP-A/CHOL and SP-A/SP-B ratios in whole surfactant harvested from subjects both at rest and immediately following exercise. Training also increased the SP-B/CHOL ratio at rest. Changes were particularly marked at rest in the SP-A/DSP, SP-A/CHOL, and SP-B/CHOL ratios in the tubular myelin-rich fraction, and after exercise in the SP-A/DSP, SP-A/CHOL, and SP-A/SP-B ratios in the tubular myelin-poor fraction. CONCLUSION: We conclude that training markedly alters the composition of alveolar surfactant both at rest and with exercise; the physiological significance of these changes remains to be determined. PMID- 11022984 TI - Heat shock proteins' mRNA expression in asthma. AB - OBJECTIVE: The aim of the present study was to investigate the expression levels of heat shock proteins (HSP) mRNA in the peripheral blood mononuclear cells (PBMC) of patients with asthma and chronic bronchitis to elucidate the role of HSP in the pathogenesis of asthma and chronic bronchitis. METHOD: Using reverse transcription-DNA polymerase chain reaction, the expression levels of HSP70, HSP90alpha and HSP90beta genes in PBMC in normal state and after heat shock were investigated. RESULTS: No HSP70 gene but HSP90alpha and HSP90beta expressions were found in non-heat-shocked PBMC of normal controls; HSP90alpha and HSP90beta genes may be expressed in PBMC of all patients, independently of acute episodes. Expression of HSP70 was found in PBMC of asthmatic patients in acute episodes and three symptom-free patients with Aas 3, step 2. Among patients with chronic bronchitis, no HSP70 gene expression was found in PBMC of patients in convalescent period but in PBMC of patients in acute episode. HSP90alpha and HSP90beta genes were expressed in PBMC of both patient groups. After heat shock, expressions of the three genes increased significantly in PBMC of both normal controls and patients. CONCLUSION: Expression of HSP70 gene in PBMC of asthmatic and chronic bronchitis patients was different, indicating that HSP, especially HSP70, might be involved in the pathogenesis of asthma. PMID- 11022986 TI - Seasonal variation in asthma hospitalizations and death rates in New Zealand. AB - OBJECTIVE: The aim of this study was to determine if there is a seasonal pattern to asthma hospitalizations and mortality in New Zealand. METHODOLOGY: Data for an 18-20 year period were obtained for the whole of New Zealand regarding the number of asthma deaths and hospital discharges in each month, as well as annual population totals (1978-1995 for asthma hospitalizations, 1976-1995 for asthma deaths). Monthly mortality and hospital discharge rates were calculated for the age groups 5-14, 15-44, and those aged 45 years and older. Variation about the mean monthly rates was plotted along a linear timescale to observe the seasonal trend. RESULTS: In the oldest age group (45+), asthma mortality and hospitalization rates peaked in the winter months (July/August). Among the younger age groups, however, mortality and hospitalizations showed different seasonal patterns; peak hospitalization occurred in the early winter months, with peak mortality in the early summer months. CONCLUSIONS: Seasonal variations in the asthma hospitalization and death rates in New Zealand are similar to those from the Northern Hemisphere. The peak mortality in summer for the younger age groups is paralleled by a reduction in hospitalizations in this period, indicating that the increase in mortality may be due to problems of access to medical care. PMID- 11022985 TI - A double-blind, placebo-controlled steroid-sparing study with budesonide Turbuhaler in Japanese oral steroid-dependent asthma patients. Japanese Pulmicort Turbuhaler study group. AB - OBJECTIVE: The aim of this study was to evaluate the oral steroid-sparing capacity of budesonide Turbuhaler. METHODOLOGY: One hundred and thirteen oral steroid-dependent patients were treated for 6 months with placebo or budesonide 800 microg or 1600 microg daily. Every second week the oral steroid dose was reduced if asthma control permitted. RESULTS: The reductions in oral steroid doses were 9, 35 and 60% in the placebo and budesonide 800 microg and 1600 microg groups, respectively. Oral steroid treatment could be discontinued in 4% (placebo), 15% (800 microg) and 23% (1600 microg). Mean peak expiratory flow values increased by 21 and 24 L/min in the budesonide groups but decreased by 6 L/min in the placebo group. Asthma attack, activity and sleep scores remained unchanged showing maintained efficacy. Plasma cortisol levels increased and an adrenocorticotropic hormone test showed improved adrenocortical response in both budesonide groups, indicating improved safety. Adverse drug reactions were infrequent and mild in all study groups. CONCLUSION: Budesonide Turbuhaler, 800 microg and 1600 microg daily, resulted in a significant reduction in oral steroid usage in steroid-dependent patients. The effect was achieved with maintained asthma control together with improvements in lung and adrenal functions. PMID- 11022988 TI - Asthma is more prevalent in rural New South Wales than metropolitan Victoria, Australia. AB - OBJECTIVE: The aim of this study was to compare the prevalence of adult respiratory symptoms and conditions in a rural setting with a metropolitan setting. METHODOLOGY: We used cross-sectional population surveys of respiratory health using the European Respiratory Health Survey screening questionnaire. A random sample of 4455 eligible young adults aged 20-44 years, from electoral rolls in south-eastern metropolitan Melbourne, Victoria, and 4521 from rural south-western New South Wales were surveyed. RESULTS: Response rates of 72% (n=3194) and 69% (n=3121) were achieved in Melbourne and the Riverina, respectively. Respondents from the Riverina reported significantly higher prevalences of nocturnal dyspnoea (P<0.01), chronic bronchitis (P=0.03), an asthma attack in the previous 12 months (P<0.001), ever having had asthma (P<0.001) and doctor-diagnosed asthma (P<0.001) compared to those from Melbourne. However, among those with 'asthma attacks in the last 12 months', Melbourne respondents experienced a higher frequency of attacks (P<0.05). Riverina respondents reported a higher prevalence of smoking (P<0.05) and smoked more cigarettes on average (P<0.001) than Melbourne respondents. However, annoyance from air pollution was higher in Melbourne than in the Riverina. CONCLUSIONS: These results suggest that asthma is more prevalent in rural southern New South Wales than in Melbourne but follows a different pattern of exacerbations. PMID- 11022987 TI - A double-blind, placebo-controlled dose-response study with budesonide Turbuhaler in Japanese asthma patients. Japanese Pulmicort Turbuhaler study group. AB - OBJECTIVE: The aim of this study was to investigate the dose-response for inhaled budesonide via Turbuhaler in Japanese patients with mild to moderate asthma. METHODOLOGY: Inhaled budesonide 100 microg, 200 microg, 400 microg or placebo was administered twice daily via Turbuhaler for 6 weeks, to 267 adult Japanese patients (mean age 51 years) with mild-to-moderate, non-steroid-dependent bronchial asthma, in a double-blind, placebo-controlled, randomized, parallel group study. The patients had to be symptomatic for more than 3 days/week and have an average morning peak expiratory flow (PEF) 50-80% of predicted normal value. RESULTS: The response to budesonide was rapid, all treatments showing a significant improvement in morning PEF after 1 week (P<0.05). During week 6, mean improvements of 15, 45, 53 and 71 L/min were observed for the placebo, 200 microg, 400 microg and 800 microg budesonide groups, respectively. Compared with placebo all improvements in the budesonide groups were statistically significant and a significant dose-response was demonstrated (P<0.001). The difference between the 200 microg and 800 microg doses was significant. Also, for several secondary efficacy variables (e.g. evening PEF, symptom score, treatment score, daily activity score and sleep score) significant dose-responses were shown. Other variables included the investigators' assessments of improvement and usefulness. They also showed statistically significant dose-response relationships and confirmed the rapid onset of action. Budesonide was well tolerated at all tested doses, with a low incidence of adverse events, all of which were minor in severity. CONCLUSIONS: Budesonide Turbuhaler in the doses 100 microg to 400 microg twice daily was effective, well tolerated and showed a rapid onset of action in patients with mild-to-moderate asthma. Dose-response was demonstrated for several variables of clinical efficacy. PMID- 11022989 TI - Diagnostic yield of bronchoscopic sampling methods in bronchial carcinoma. AB - OBJECTIVES: The aim was to study the yield of malignant or suspicious cells from bronchoalveolar lavage (BAL), endobronchial biopsy (BX) and endobronchial brushing (BR) specimens obtained at bronchoscopy. METHODOLOGY: We prospectively followed up all patients who had undergone bronchoscopy and studied those with subsequent diagnosis of bronchial carcinoma at a tertiary referral centre. Bronchoalveolar lavage, BX and BR were performed, sequentially whenever possible, by one pulmonologist. The parameters assessed were age, gender, macroscopic bronchoscopic findings, TNM staging, radiological findings, histological typing, and diagnostic yield from BAL, BX, and BR. RESULTS: One hundred patients (31 females, 69 males; mean age +/- SD, 61.8+/-12.7 years; range 32-81 years) were studied between 1995 and 1997. Of these, BAL, BX and BR were performed on 100, 64 and 37 cases which yielded diagnostic specimens for bronchial carcinoma in 69, 78.1, and 62.2% of cases, respectively (P > 0.05). Diagnostic specimens were therefore obtained in 69,50, and 23% of the entire patient cohort from BAL, BX, and BR, respectively (P < 0.001). The diagnostic yield of BAL was independent of patient age, gender, site of lesion, TNM staging, histological typing and macroscopic bronchoscopic findings. Addition of BX, BR and postbronchoscopic sputum sampling to BAL only increased the yield of diagnostic specimens for bronchial carcinoma by 7, 0, and 4%, respectively. CONCLUSION: Bronchoalveolar lavage is a highly effective sampling method to obtain cytological evidence for bronchial carcinoma. The diagnostic yield for BAL is independent of tumour or other clinical characteristics. Further studies should be performed to confirm these important and clinically relevant findings. PMID- 11022990 TI - Acoustic parameters of voluntary cough in healthy non-smoking subjects. AB - OBJECTIVE: The aim of this study was to explore cough in healthy subjects. METHODOLOGY: We studied 234 coughs generated by 24 (12 males) healthy non-smokers (forced expiratory volume in 1 s (FEV1) 103+/-8% of predicted), who had no significant differences in FEV1 and age between males and females. For each subject, several bouts of voluntary coughing were recorded using a personal computer with an A/D converter (sampling rate 10 kHz, 8 bit resolution) and the first and second coughs of each bout were analysed using short-time Fast Fourier Transformation. For each cough we studied the three phases that are produced. In particular, we studied the duration of the three parts, loudest frequency in the first part, lowest and highest frequencies, number of continuous frequencies and lowest and highest continuous frequencies in the second part, and the loudest frequency of the third part if present. RESULTS: We found significant differences between males and females in length of the first part (41.4+/-14 vs 44.7+/-10.4 msec, P = 0.04), loudest frequency of the first part (362+/-145 vs 449+/-145 Hz), lowest frequencies (282+/-100 vs 348+/-135 Hz) and highest continuous frequencies (3877+/-571 vs 4147+/-362 Hz; P < 0.001) of the second part. An interesting finding was that healthy males and females had the same number of continuous frequencies. Different frequencies are probably a consequence of anatomical differences in airway geometry involved in the cough. CONCLUSION: In cough frequency spectrum studies the differences between the two sexes should be taken into account to reduce the variability of the results. PMID- 11022991 TI - Primary spontaneous pneumothorax: treatment practices in Australia. AB - OBJECTIVE: The aim of the present study was to determine treatment practices for spontaneous pneumothorax (PTX) in Australia. METHODOLOGY: A questionnaire regarding treatment of PTX was posted to all medically qualified members of the Thoracic Society of Australia and New Zealand resident in Australia. RESULTS: Fifty-three per cent (n = 226) of questionnaires were returned. Twenty-one responses were judged to be informal and rejected from analysis. The results indicate a general agreement in the treatment of small and large PTX but a wide variation in PTX of intermediate size. CONCLUSION: Opinion regarding the optimal treatment for PTX of intermediate size was found to vary considerably. There is a need for evidence-based development and publication of best practice guidelines for PTX. PMID- 11022992 TI - Management and treatment perceptions among young adults with asthma in Melbourne: the Australian experience from the European Community Respiratory Health Survey. AB - OBJECTIVES: As part of the European Community Respiratory Health Survey (ECRHS) in 1992-1993 we assessed management practices and treatment perceptions among young asthmatic adults in Melbourne, Australia. METHODOLOGY: We conducted a postal questionnaire survey of 4500 randomly selected adults (aged 20-44 years), drawn from three electoral districts, of whom 3200 (71%) subjects responded. A randomly selected sample of 1642 respondents, 'enriched' by a further 433 symptomatic subjects, was invited to complete a second phase respiratory questionnaire. RESULTS: The questionnaire was completed by 757 subjects who underwent laboratory testing. A further 119 subjects who were unable to attend the laboratory completed an identical questionnaire by telephone interview (42% response rate). In the second phase, 16% of subjects reported 'current asthma' (group I) as defined by physician confirmation and a recent attack (within 1 year), 10% had confirmed asthma but reported no recent attack (group II) and 74% did not have asthma (group III). Inhaled corticosteroid use was significantly higher in group I than in group II subjects (45% vs 24%, P<0.01), but only 11% of asthmatic subjects overall reported daily prophylactic use. Regular treatment in any form was considered to be 'bad' by 65% of asthmatic subjects and only 43% took medication as prescribed all of the time. CONCLUSION: Despite national education campaigns, the majority of young asthmatic adults in Melbourne did not adhere to prescribed treatment, but continued to rely upon beta2-agonists alone with neglect of regular inhaled corticosteroid which has probably contributed to Australia's continued high asthma morbidity and mortality rates. PMID- 11022993 TI - Changes in malondialdehyde levels in bronchoalveolar fluid and serum by the treatment of asthma with inhaled steroid and beta2-agonist. AB - OBJECTIVE: Oxidative stress plays an important role in the pathogenesis of asthma. Recent data suggest that clinical indices of the patients with asthma may not correlate with the underlying inflammatory process. We aimed to measure the level of malondialdehyde (MDA), which is a marker of lipid peroxidation, a free radical-mediated process, before and after a well-accepted treatment of asthma. METHODOLOGY: Nine non-smoking females and five non-smoking males with mild moderate asthma were included. Twenty-four age- and sex-matched, non-smoking healthy people (17 females and seven males, mean age 32.1 years, range 20-59) were included for control. After initial evaluation, spirometry, bronchoscopy with bronchoalveolar lavage (BAL), and blood sample were maintained. The patients were treated with twice-daily salmeterol inhaler (100 microg/d) and fluticasone propionate inhaler (500 microg/d). One month later the investigations were repeated. Serum MDA levels before treatment were compared with both the levels after treatment and levels of controls. Malondialdehyde levels of BAL were compared before and after treatment. RESULTS: Serum MDA level of the patient before treatment was 6.7+/-0.8 nmol/mL, significantly higher than that of healthy controls; 3.8+/-0.4, P < 0.001. One month after the treatment, serum MDA level decreased to 5.3+/-0.7 nmol/mL (P < 0.001). However, this level is still significantly higher than healthy controls (P < 0.0001). Forced expiratory volume in 1 s level of the patients increased from 2.43+/-0.79 L to 3.50+/-1.21 L after the treatment (P < 0.001). CONCLUSION: Although treatment with beta2-agonist and corticosteroid inhalers for the duration of 1 month reduced lipid peroxidation significantly, it was still at a level significantly higher than healthy controls. The treatment may need a longer duration to improve lipid peroxidation or an alternative regimen which is more effective in controlling inflammation may be warranted. PMID- 11022994 TI - Endobronchial actinomycosis associated with a foreign body. AB - Primary endobronchial actinomycosis is an exceedingly rare condition which closely mimics bronchial carcinoma both clinically and radiologically. We report the case of a middle-aged Chinese man who had right lower lobe endobronchial actinomycosis associated with inhalation of a presumed chicken bone. The patient made a complete clinical and radiological recovery after a 2 month combined course of amoxycillin and clavulanate, after removal of the foreign body, which was only detected after using contiguous computed tomography. Our experience in the management of this patient should help clinicians in the investigation and management of this rare but treatable condition. PMID- 11022995 TI - A case of pulmonary tumour embolism mimicking miliary tuberculosis. AB - Pulmonary metastases from choriocarcinoma can very rarely give rise to a 'miliary' pattern on the chest X-ray. A 23-year-old woman with a diffuse nodular pattern on chest X-ray died due to acute respiratory failure and cor pulmonale. At autopsy, choriocarcinoma of the uterus was found. In the lungs, multiple macrovascular tumour emboli within branches of pulmonary muscular arteries in the region of segmental/subsegmental bronchi were detected. There was no evidence of pulmonary parenchymal metastases, pneumonia or tuberculosis. The 'miliary' pattern of pulmonary metastases described in cases of choriocarcinoma may be due to large vessel intra-arterial tumour emboli rather than disseminated parenchymal metastases. PMID- 11022996 TI - Chylothorax occurring 23 years post-irradiation: literature review and management strategies. AB - Chylothorax is an uncommon condition rarely seen outside the clinical scenario of trauma or malignancy, and management may be difficult. We report the case of a 52 year-old man with a right chylothorax occurring 23 years after mantle irradiation for Hodgkin's disease. In addition, a Medline literature search was performed. Chylothorax occurring post-irradiation is rare and the aetiology is uncertain. Recurrent malignancy must be excluded. There are a number of management options and our patient was successfully treated with a combination of thoracocentesis, medium chain triglyceride diet and thoracoscopy with talc pleurodesis. There was no evidence of recurrent chylothorax at 2 years follow up. Prompt assessment and treatment is required. Surgical measures need to be considered early if medical treatment is unsuccessful. PMID- 11022997 TI - A case of idiopathic constrictive bronchiolitis in a middle-aged male smoker. AB - When one sees a middle-aged male smoker who presents with progressive exertional dyspnoea and irreversible airflow obstruction, the most likely clinical diagnosis is pulmonary emphysema or chronic obstructive pulmonary disease (COPD). We report a 45-year-old male smoker who was initially suspected to have such a disease but was eventually diagnosed as having idiopathic constrictive bronchiolitis by lung biopsy, clinical history, and laboratory findings. A finding on lung computed tomography of diffuse hyperinflation but few low attenuation areas and relatively well-preserved diffusing capacity of carbon monoxide seems to be the key for suspecting this rare clinical entity. The pathological difference between this bronchiolitis and small airway disease observed in COPD will be also discussed. PMID- 11022998 TI - Introduction: nutritional and functional roles of eggs in the diet. PMID- 11022999 TI - The changing face of functional foods. AB - Consumers began to view food from a radically different vantage point in the 1990s. This 'changing face' of food has evolved into an exciting area of the food and nutrition sciences known as functional foods. Functional foods can be defined as those providing health benefits beyond basic nutrition and include whole, fortified, enriched or enhanced foods which have a potentially beneficial effect on health when consumed as part of a varied diet on a regular basis at effective levels. Interest in functional foods skyrocketed in the last decade due to a number of key factors, including the growing self-care movement, changes in food regulations and overwhelming scientific evidence highlighting the critical link between diet and health. The interest in functional foods has resulted in a number of new foods in the marketplace designed to address specific health concerns, particularly as regards chronic diseases of aging. In addition to new foods designed specifically to enhance health, however, functional foods can also include those traditional, familiar foods for which recent research findings have highlighted new health benefits or dispelled old dogma about potential adverse health effects. An excellent example is the American egg-Nature's original functional food. Eggs have not traditionally been regarded as a functional food, primarily due to concerns about their adverse effects on serum cholesterol levels. Furthermore, it is now known that there is little if any connection between dietary cholesterol and blood cholesterol levels and consuming up to one or more eggs per day does not adversely affect blood cholesterol levels. Finally, eggs are an excellent dietary source of many essential (e.g., protein, choline) and non-essential (e.g., lutein/zeaxanthin) components which may promote optimal health. Nutrition in the new millennium will be dramatically different than it was in the 20th century. Completion of the human genome project will facilitate the identification of humans predisposed to diet-related diseases. Targeted or 'prescription' nutrition will become the norm, enabling the food and medical industries to provide timely and individualized approaches to disease prevention and health promotion. The egg will continue to play an important role in the changing face of functional foods. PMID- 11023000 TI - The functional food trend: what's next and what Americans think about eggs. AB - OBJECTIVE: The Health Focus National Study of Public Attitudes and Actions Toward Healthy Foods is conducted every two years to identify current issues in consumer health and nutrition behavior and attitudes, to assess the trends in consumer priorities and to develop an understanding of where consumers are headed in their behavior towards their health and diet. This paper focuses on consumer interests in functional nutrition for disease prevention and health enhancement. It examines the role consumers see for eggs in healthy diets. METHODS: The data for this study were collected from written questionnaires completed by 2,074 qualified respondents in August, September and October of 1998. The research was conducted in two stages: 1. A telephone pre-recruit from a national probability sample of households qualified respondents as Primary Grocery Shoppers (those who make most of the food buying decisions for their household or who equally share that responsibility). 2. A 12-page, self-administered questionnaire was mailed to qualified respondents. Respondents to this survey represent shoppers in the U.S. in all respects except race. Women account for 81% of the survey respondents, since they do most of the household shopping. RESULTS: Most shoppers believe foods can offer benefits that reach beyond basic nutrition to functional nutrition for disease prevention and health enhancement. As consumers better understand the functional benefits of eggs, from Prevention to Performance, Wellness, Nurturing and Cosmetics, eggs will continue to play an important role in healthy eating for many consumers. CONCLUSIONS: Eggs are considered a healthy food by most consumers as long as they are eaten in moderation. Increased egg consumption is being driven by consumer interest in health benefits that reach beyond dietary avoidance strategies to positive nutrition strategies. Today's self-reliant approach to health creates significant opportunities for health and nutrition marketers to use knowledge-based marketing programs to shape present and future health decisions and product choices among shoppers. PMID- 11023001 TI - Beyond the zone: protein needs of active individuals. AB - There has been debate among athletes and nutritionists regarding dietary protein needs for centuries. Although contrary to traditional belief, recent scientific information collected on physically active individuals tends to indicate that regular exercise increases daily protein requirements; however, the precise details remain to be worked out. Based on laboratory measures, daily protein requirements are increased by perhaps as much as 100% vs. recommendations for sedentary individuals (1.6-1.8 vs. 0.8 g/kg). Yet even these intakes are much less than those reported by most athletes. This may mean that actual requirements are below what is needed to optimize athletic performance, and so the debate continues. Numerous interacting factors including energy intake, carbohydrate availability, exercise intensity, duration and type, dietary protein quality, training history, gender, age, timing of nutrient intake and the like make this topic extremely complex. Many questions remain to be resolved. At the present time, substantial data indicate that the current recommended protein intake should be adjusted upward for those who are physically active, especially in populations whose needs are elevated for other reasons, e.g., growing individuals, dieters, vegetarians, individuals with muscle disease-induced weakness and the elderly. For these latter groups, specific supplementation may be appropriate, but for most North Americans who consume a varied diet, including complete protein foods (meat, eggs, fish and dairy products), and sufficient energy the increased protein needs induced by a regular exercise program can be met in one's diet. PMID- 11023002 TI - The potential role of dietary xanthophylls in cataract and age-related macular degeneration. AB - The carotenoid xanthophylls, lutein and zeaxanthin, accumulate in the eye lens and macular region of the retina. Lutein and zeaxanthin concentrations in the macula are greater than those found in plasma and other tissues. A relationship between macular pigment optical density, a marker of lutein and zeaxanthin concentration in the macula, and lens optical density, an antecedent of cataractous changes, has been suggested. The xanthophylls may act to protect the eye from ultraviolet phototoxicity via quenching reactive oxygen species and/or other mechanisms. Some observational studies have shown that generous intakes of lutein and zeaxanthin, particularly from certain xanthophyll-rich foods like spinach, broccoli and eggs, are associated with a significant reduction in the risk for cataract (up to 20%) and for age-related macular degeneration (up to 40%). While the pathophysiology of cataract and age-related macular degeneration is complex and contains both environmental and genetic components, research studies suggest dietary factors including antioxidant vitamins and xanthophylls may contribute to a reduction in the risk of these degenerative eye diseases. Further research is necessary to confirm these observations. PMID- 11023003 TI - Choline: needed for normal development of memory. AB - Choline is a dietary component essential for normal function of all cells. It, or its metabolites, assures the structural integrity and signaling functions of cell membranes; it is the major source of methyl-groups in the diet (one of choline's metabolites, betaine, participates in the methylation of homocysteine to form methionine); and it directly affects nerve signaling, cell signaling and lipid transport/metabolism. In 1998, the National Academy of Sciences, USA, issued a report identifying choline as a required nutrient for humans and recommended daily intake amounts. Eggs are an excellent dietary source of choline. Pregnancy and lactation are periods when maternal reserves of choline are depleted. At the same time, the availability of choline for normal development of the brain is critical. When rat pups received choline supplements (in utero or during the second week of life), their brain function changed, resulting in the lifelong memory enhancement. This change in memory function appears to be due to changes in the development of the memory center (hippocampus) in the brain. The mother's dietary choline during a critical period in brain development of her infant influences the rate of birth and death of nerve cells in this center. These changes are so important that we can pick out the groups of animals whose mothers had extra choline even when these animals are elderly. Thus, memory function in the aged rat is, in part, determined by what the mother ate. This is not the first example of a critical nutrient that must be present at a specific time in brain development. If folate isn't available in the first few weeks of pregnancy, the brain does not form normally. Thus, we suggest that pregnancy is a period when special attention has to be paid to dietary intake. PMID- 11023004 TI - The symbolization of eggs in American culture: a sociologic analysis. AB - OBJECTIVE: To demonstrate that the recent decline in egg consumption in the United States was, in part, the result of a food scare that began in the 1960's. METHODS: Using the Reader's Guide to Periodical Literature, the frequency of articles about eggs, dietary cholesterol and heart disease in popular magazines was obtained. A content analysis was performed on a random sample of these articles. RESULTS: The increased trend in magazine articles and public statements by groups such as the American Heart Association linking eggs, blood cholesterol and heart disease is associated with the downward trend in egg consumption. CONCLUSIONS: Public exposure to negative messages about particular foods can contribute to a decline in their consumption. Exposure to more consistently positive messages about foods can bring about an increase in the consumption of those foods. PMID- 11023005 TI - The impact of egg limitations on coronary heart disease risk: do the numbers add up? AB - For over 25 years eggs have been the icon for the fat, cholesterol and caloric excesses in the American diet, and the message to limit eggs to lower heart disease risk has been widely circulated. The "dietary cholesterol equals blood cholesterol" view is a standard of dietary recommendations, yet few consider whether the evidence justifies such restrictions. Over 50 years of cholesterol feeding studies show that dietary cholesterol does have a small effect on plasma cholesterol concentrations. The 167 cholesterol feeding studies in over 3,500 subjects in the literature indicate that a 100 mg change in dietary cholesterol changes plasma total cholesterol by 2.2 mg/dL. Today we recognize that dietary effects on plasma cholesterol must be viewed from effects on the atherogenic LDL cholesterol as well as anti-atherogenic HDL cholesterol since the ratio of LDL:HDL cholesterol is a major determinant of heart disease risk. Cholesterol feeding studies demonstrate that dietary cholesterol increases both LDL and HDL cholesterol with little change in the LDL:HDL ratio. Addition of 100 mg cholesterol per day to the diet increases total cholesterol with a 1.9 mg/dL increase in LDL cholesterol and a 0.4 mg/dL increase in HDL cholesterol. On average, the LDL:HDL ratio change per 100 mg/day change in dietary cholesterol is from 2.60 to 2.61, which would be predicted to have little effect on heart disease risk. These data help explain the epidemiological studies showing that dietary cholesterol is not related to coronary heart disease incidence or mortality across or within populations. PMID- 11023006 TI - Egg consumption and coronary heart disease: an epidemiologic overview. AB - Serum cholesterol has been established as a modifiable risk factor for coronary heart disease. Experimental feeding studies show that saturated fat and cholesterol increase serum cholesterol levels; thus, dietary recommendations for lowering the risk of heart disease proscribe the intake of both substances. Recommendations have also included limits on the intake of eggs because of their high cholesterol content. In free-living populations, diet reflects a pattern of associated choices. Increases in one food may lead to changes in the consumption of other foods that may modulate disease risk. Epidemiologic data are helpful in assessing the importance of foods and nutrients in the context in which they are actually consumed. We review epidemiologic data relating dietary cholesterol and eggs to coronary disease risk. Cholesterol intake was associated with a modest increase in the risk of coronary events. The true magnitude of the association is difficult to estimate because most studies fail to account for potential confounding by other features of the diet. When a full-range of confounding factors was considered, the association between cholesterol intake and heart disease risk was small (6% increase in risk for 200mg/1,000kcal/day difference in cholesterol intake). Several studies have examined egg intake and its relationship with coronary outcomes. All but one failed to consider the role of other potentially confounding dietary factors. When dietary confounders were considered, no association was seen between egg consumption at levels up to 1 + egg per day and the risk of coronary heart disease in non-diabetic men and women. PMID- 11023007 TI - Nutritional contribution of eggs to American diets. AB - OBJECTIVES: The main purposes of this study were (1) to assess the nutritional significance of eggs in the American diet and (2) to estimate the degree of association between egg consumption and serum cholesterol concentration. METHODS: Data from the most recent National Health and Nutritional Examination Survey (NHANES III, 1988-94) were utilized to compare the nutritional quality indicators of diets that contained eggs (USDA food grouping system) with those that did not. Nutrient intake (from 24-hour dietary recall), egg intake (from food frequency questionnaire), sociodemographic data and blood cholesterol levels of subjects who met inclusion criteria (n = 27,378) were grouped according to the occurrence and frequency of egg consumption and were analyzed using SUDAAN. RESULTS: Daily nutrient intake of egg consumers (EC) was significantly greater than that of nonconsumers (NC) for all nutrients studied (except dietary fiber and vitamin B6). Eggs contributed < 10% of daily intake of energy and vitamin B6, 10% to 20% of folate and total, saturated and polyunsaturated fat, and 20% to 30% of vitamins A, E and B12 in EC. Compared to EC, NC had higher rates of inadequate intake (defined by Estimated Average Requirements (EAR) or < 70% Recommended Dietary Allowance (RDA)) for vitamin B12 (10% vs. 21%), vitamin A (16% vs. 21%), vitamin E (14% vs. 22%) and vitamin C (15% vs. 20%). After adjusting for demographic (age, gender and ethnicity) and lifestyle variables (smoking and physical activity), dietary cholesterol was not related to serum cholesterol concentration. People who reported eating > or = 4 eggs/wk had a significantly lower mean serum cholesterol concentration than those who reported eating < or = 1 egg/wk (193 mg/dL vs. 197 mg/dL, p < 0.01). More frequent egg consumption was negatively associated with serum cholesterol concentration (beta = -6.45, p < 0.01). CONCLUSIONS: In this cross-sectional and population-based study, egg consumption made important nutritional contributions to the American diet and was not associated with high serum cholesterol concentrations. PMID- 11023008 TI - Individual differences among cocaine users. AB - The present study examined whether individual differences in personality could differentiate two types of cocaine users. We hypothesized that self-medicators (SM) use cocaine as a way to alleviate their dysphoric moods, whereas sensation seekers (SS), in contrast, use cocaine primarily to engender positive mood states. Eighteen male cocaine users were classified based on two dimensions of the Tridimensional Personality Questionnaire. SM were defined by having high harm avoidance (>17) and low novelty-seeking scores (<18), and SS by high novelty seeking (>18) and low harm-avoidance scores (<17). It was predicted that SM would report higher depression and anxiety than would SS, and would also exhibit a brain activity pattern similar to that found in clinical depression. The results showed that SM reported higher anxiety than SS, F(1, 8) = 27.5, p < .001, but did not differ in depression. SM exhibited decreased blood flow within the left frontal lobes, F(1, 10) = 6.78, p < .05, similar to what has been observed in major depressive disorder. These findings suggest the importance of attending to individual differences in the motivation for cocaine use so that treatment can be targeted more effectively. PMID- 11023009 TI - Relationships between sensation seeking and emotional symptomatology during smoking cessation with nicotine patch therapy. AB - This study explored relationships between the sensation-seeking trait and the development of emotional symptomatology during smoking cessation with nicotine transdermal patches. Twenty-five subjects were evaluated before they stopped smoking, on Day 8, Day 30, Day 90, and Day 120. Initial motives for smoking and the sensation-seeking personality trait were tested as possible predictors for the development of specific mood disturbances. Our subjects scored very high on sensation seeking, consistent with previous results on smokers. This may also be due to the well-known tendency of high sensation-seekers to be willing to try new experiences. The sensation-seeking trait did not predict the issue of cessation. However, it was related to emotional deficit (anhedonia, affective blunting), tiredness, and a lack of energy, before and during smoking cessation. Two different interpretations of emotional deficit are proposed. PMID- 11023010 TI - Self-assessment of inebriation from external indices. AB - OBJECTIVE: The objective was to study people's judgments, through the use of external indices, of their expected degree of inebriation and to characterize the cognitive processes involved in making these judgments. METHOD: The participants were 143 adults living in France. In accordance with N. H. Anderson's functional theory of cognition (1982, 1996), their primary task was to read 64 cards depicting drinking situations characterized by three parameters: number of glasses, type of drink, and temporal relation to eating; to associate with each situation a certain degree of personal intoxication; and to indicate this degree of expected intoxication on linear scale anchored by Not at all drunk and Extremely drunk. RESULTS: Participants were able to judge the general direction and combination of effects. They indicated incorrectly, however, that incremental effects decrease at higher levels of consumption (4 or more glasses) and at higher concentrations of alcohol in a drink. On the other hand, they indicated correctly that eating food decreases the impact of drinking alcohol. Drinkers and nondrinkers used different rules for self-assessment from external indices. IMPICATIONS: The systematic errors in both drinkers' and nondrinkers' use of external clues could, unless corrected, result in their becoming more inebriated than intended. PMID- 11023011 TI - Screening of drug use in a teenage Brazilian sample using the Drug Use Screening Inventory (DUSI). AB - The Brazilian translation of the Drug Use Screening Inventory (DUSI) was applied to 213 Brazilian teenagers who were classified according to their alcohol and/or drug dependence level (DSM-III-R) as: 71 nondrug users (Group 1), 71 with light/moderate dependence (Group 2) and 71 with severe dependence (Group 3). The DUSI was applied and the absolute density in each of 10 areas was calculated. The three groups presented statistically significant differences (p < .001) in the "substance use" area, with the following values (medians +/- interquartile range): Group 1: 0+/-7; Group 2: 20+/-33 and Group 3: 80+/-33. The groups also presented significant differences in behavior pattern, social competency, family system, work adjustment, peer relationships and leisure/recreation. Other differences detected among the groups indicated an important relationship between drug use and school delay. A good Spearman rank correlation (0.86, p < .0001) was observed between Composite International Diagnostic Interview (CIDI) diagnosis and DUSI, indicating that this instrument can be useful in the screening of substance use among Brazilian teenagers. PMID- 11023012 TI - Smoking behavior among casino employees: self-report validation using plasma cotinine. AB - The veracity of behavioral self-reports is often challenged, particularly when the motivation to avoid stigma and win social approval holds potential to introduce bias into the data collected. This study employed plasma cotinine tests to validate the self-reports of tobacco use collected from 3,841 casino employees as part of a comprehensive health survey. Rates of discordance were calculated by comparing employee self-reports with results from plasma colinine tests. This study provides evidence that casino employees can provide valid self-report data. Further, discordance rates of self-reported tobacco use vary according to operational definitions of tobacco use. These findings highlight the methodological importance of recognizing the inherent heterogeneity of smoking behavior. PMID- 11023013 TI - Hispanic and Anglo college women's risk factors for substance use and eating disorders. AB - This study investigated the role of temperament style (Novelty Seeking and Harm Avoidance) of Hispanic American and Anglo college women in moderating and mediating the relationship between family addiction/family functioning and offspring problem behaviors. The sample was comprised of 67 Hispanic American and 770 Anglo undergraduate women. Findings of this study indicate that the processes of risk that lead to substance use and eating disorders follow different routes for Hispanic American and Anglo women. Novelty Seeking and Harm Avoidance were found to be important factors in both moderating and mediating the effects of parental drinking and family dysfunction for both Hispanic and Anglo college women. PMID- 11023014 TI - Eight-year follow-up of a community-based large group behavioral smoking cessation intervention. AB - OBJECTIVE: To evaluate the effectiveness of a large group community-based behavioral smoking cessation intervention over an 8-year follow-up period and to determine precessation predictors of cessation at each follow-up time. RESEARCH APPROACH: Behavioral intervention followed by three longitudinal follow-up interviews. SETTING: Regional Outpatient Cancer Centre. STUDY PARTICIPANTS: 971 participants in smoking cessation clinics held between 1986 and 1990. INTERVENTION: Eight 90-minute sessions over 4 months utilizing education, self monitoring, nicotine fading, a group quit date and behavioral modification techniques. Up to 110 smokers participated in each group program. MAIN OUTCOME MEASURES: Cessation rates at 3, 6, and 12 months postquit and at 8-year follow up. Differences between successful and unsuccessful participants in precessation demographic, smoking history, and smoking behavior variables. RESULTS: At 3 months postquit date, 39.3% of the 971 participants reported that they were not smoking, decreasing to 32.1% at 6 months and 26.0% at 12 months. At the 8-year follow-up, 33.9% of the original sample were contacted, and of those, 47.7% reported that they were currently not smoking. There were nine predictors of cessation at the end of the program (3 months), which were similar to those previously reported in the literature. Similarly, at 6 and 12 months, six factors were associated with not smoking. At the 9-year follow-up the only variable predictive of continued abstinence was being female (p < .05). CONCLUSIONS: This program was successful in promoting smoking cessation and maintenance, even with its large-group format. Predictive factors were similar to those previously reported in the literature. PMID- 11023015 TI - To thine own self be true: self-concept and motivation for abstinence among substance abusers. AB - Individuals approved for public-sector addiction treatment were interviewed regarding their reasons for attempting abstinence. Follow-up interviews were completed 3 to 6 months after participants' removal from county-controlled treatment wait-lists. Rates of continuous self-reported abstinence for 90 days preceding follow-up were positively associated with motivation linked to discrepancies between substance use and self-standards. Characteristics associated with high identity-linked motivation were cocaine preference, a history of reducing self-dissatisfaction through substance use, low rewards and high costs associated with using. and low support for the user identity among significant others. The perception of discrepancies between substance use and self-standards was an effective motivator of abstinence even among those who reported previous use of substances to dampen self-dissatisfaction. PMID- 11023016 TI - Weight concerns in women smokers during pregnancy and postpartum. AB - When women smokers become pregnant, they are asked to control weight gain and at the same time to relinquish an addictive drug with weight suppressing effects. For women with serious body image concerns or a history of unhealthful eating patterns, smoking cessation may be particularly problematic. To investigate the relationship of weight concerns with smoking and weight gain during pregnancy and postpartum weight loss strategies, we conducted a retrospective study of women who had given birth to their first child within the past 10 years and were smokers when they became pregnant. We observed that women smokers with high weight and body image concerns (HC) gained significantly more weight during pregnancy-in amounts that far exceeded maximum recommended weight gain-than did women with low concerns (LC). HC were more likely to adopt smoking as a weight control strategy and to be receptive to multiple weight-control strategies. Although they lost significantly more weight in the first month postpartum than did LC, they had also gained significantly more during pregnancy; the net result was that weight loss as a percentage of weight gained did not differ significantly between groups. HC were significantly less likely to experience food cravings in the first trimester and marginally less likely to vomit than LC. We conclude that early identification of high-risk women, coordination of prenatal care with smoking cessation counseling, and development of effective relapse prevention strategies that specifically address weight issues both during and after pregnancy will be needed if efforts to reduce smoking during pregnancy and postpartum are to be optimized. PMID- 11023017 TI - Preventing illicit drug use in adolescents: long-term follow-up data from a randomized control trial of a school population. AB - National survey data indicate that illicit drug use has steadily increased among American adolescents since 1992. This upward trend underscores the need for identifying effective prevention approaches capable of reducing the use of both licit and illicit drugs. The present study examined long-term follow-up data from a large-scale randomized prevention trial to determine the extent to which participation in a cognitive-behavioral skills-training prevention program led to less illicit drug use than for untreated controls. Data were collected by mail from 447 individuals who were contacted after the end of the 12th grade, 6.5 years after the initial pretest. Results indicated that students who received the prevention program (Life Skills Training) during junior high school reported less use of illicit drugs than controls. These results also support the hypothesis that illicit drug use can be prevented by targeting the use of gateway drugs such as tobacco and alcohol. PMID- 11023018 TI - Eating and body image concerns among obese and average-weight children. AB - Research compared obese and average-weight children with regard to concerns about being or becoming overweight, history of dieting, concerns about the effects of eating food, and perceived discrepancy between real and ideal body image. Participants included 526 obese and average-weight elementary-age school children to whom questionnaires were administered. Gender (male/female), obesity status (obese/average-weight), and grade level (lower elementary/upper elementary) were considered. Obese children were significantly more likely to engage in dieting behaviors, to express concern about their weight, to restrain their eating, and to exhibit more dissatisfaction with their body image than average-weight children. Girls were more likely to exhibit these behaviors than were boys. These findings suggest the importance of studying the emergence of disordered eating habits in childhood. PMID- 11023019 TI - The co-occurrence of smoking and binge drinking in adolescence. AB - The present work sought to determine adolescent rates of smoking and binge drinking co-occurrence. Secondary analyses were conducted on the interview responses of more than 4,000 adolescents between the ages of 13 and 18 who took part in the 1995 National Household Survey on Drug Abuse. Results revealed that not only are adolescent smokers likely to be binge drinkers, but adolescent binge drinkers are also likely to be smokers. Conversely, those who abstain from involvement with one of these substances generally abstain from the other as well. The discussion highlighted race/ethnic and gender differences in co occurrence rates as well as prevention and intervention implications. PMID- 11023020 TI - Characteristics of research volunteers for inpatient cocaine studies: focus on selection bias. AB - In order to investigate the selection bias of subjects for inpatient human cocaine studies, characteristics of 859 potential subjects were examined. Excluded subjects compared with accepted group were more likely to be single and male, currently use drugs other than cocaine, have a history of intravenous cocaine use, and have medical or mental health problems or physical complaints. Subjects who were accepted but did not participate, compared with participants, were likely to spend more money on cocaine. These results suggest that potential subjects who were accepted to our research studies may not accurately represent all potential subjects for several important subject characteristics. PMID- 11023021 TI - Impulsiveness and compulsiveness in alcoholics and nonalcoholics. AB - Impulsiveness and compulsiveness questionnaires were completed by recovering alcoholics (n = 54) and by a community sample (n = 351). No relationship was found, indicating these traits are independent, distinct constructs. Alcoholics scored significantly higher on impulsiveness than nonalcoholics, and impulsiveness was negatively associated with age for both groups. Interestingly, the significant difference between the two groups on impulsiveness was maintained across age groups. Alcoholic and nonalcoholic compulsiveness scores showed no difference; however, compulsiveness was negatively related to age in nonalcoholics, but not alcoholics. PMID- 11023022 TI - Childhood trauma as a correlate of lifetime opiate use in psychiatric patients. AB - Associations have been reported between childhood sexual and/or physical abuse and adult substance abuse in general. This study investigated the relationship between childhood abuse and opiate use in particular among 763 consecutively admitted psychiatric inpatients. Patients were interviewed about demographic information, alcohol and drug use, and history of interpersonal violence. Opiate users were 2.7 times more likely to have a history of childhood sexual and/or physical abuse than nonopiate users, after controlling for diagnostic and sociodemographic variables. Opiate use was higher among those reporting physical abuse alone (24.1%) or both physical and sexual abuse (27%) than among those reporting sexual abuse alone (8.8%). Implications of these findings are discussed. PMID- 11023023 TI - Palliative analgesic effect of Re-186 HEDP in various cancer patients with bone metastases. AB - The clinical picture of bone metastases is manifested by pain and loss of mechanical stability. Standard treatment options for bone metastases include external beam radiotherapy and the use of analgesics. Due to a large number of lesions in many patients, the use of radionuclide therapy with beta emitters may be preferable. Re-186 hydroxyethylidene diphosphonate (Re-186 HEDP) is one of the radiopharmaceuticals suitable for palliative treatment of metastatic bone pain. The aim of this study was to investigate palliative and side effects of Re-186 HEDP in patients with different types of cancers. MATERIAL & METHOD: Thirty one (17 male, 14 female) patients with various cancers (10 prostate, 10 breast, 4 rectum, 5 lung, 2 nasopharynx) and bone metastases were included in the study. Therapy was started with a fixed dose of 1295 MBq of Re-186 HEDP. If necessary, the same dose was repeated at least 3 times after an interval of 10-12 weeks; A total of 40 standard doses were given; 6 patients received repeated doses (3 doses in 3 patients, 2 doses in 3 patients). The patients with bone marrow suppression were excluded from the study. The pain relief was assessed the Eastern Cooperative Oncologic Group (ECOG) and the Karnofsky status index. All patients were evaluated with standard evaluation forms filled in daily for a maximum of 10 weeks. RESULTS: The mean response rate was 87.5% in patients with breast and prostate cancer, 75% in patients with rectum cancer and 20% in patients with lung cancer. The overall response rate was 67.5%. The palliation period varied between 6 and 10 weeks, with a mean of 8.1+/-1.3 weeks. The maximal palliation effect was observed between the 3rd and 7th weeks. No serious side effects were seen except mild hematologic toxicity. DISCUSSION & CONCLUSION: It is concluded that Re-186 HEDP is a highly effective agent in the palliation of metastatic bone pain in patients with prostate, breast and rectum cancer, but not effective in lung cancer. On the other hand, Re-186 seems to be a good alternative to Sr-89 because of its preferable physical characteristics (such as short half life and gamma energy emission), low side effect profile, early response and repeatability. PMID- 11023024 TI - Evaluation of iodinated and brominated [11C]styrylxanthine derivatives as in vivo radioligands mapping adenosine A2A receptor in the central nervous system. AB - In vivo assessment of the adenosine A2A receptors localized in the striatum by PET or SPECT offers us a new diagnostic tool for neurological disorders. In the present study, we evaluated the potential of iodinated and brominated styrylxanthine derivatives labeled with 11C as an in vivo probe. [7-Methyl-11C] (E)-3,7-dimethyl-8-(3-iodostyryl)-1-propargylxan thine ([11C]IS-DMPX) and [7 methyl-11C]-(E)-8-(3-bromostyryl)-3,7-dimethyl-1-propargylxa nthine ([11C]BS DMPX) were prepared by the 11C-methylation of corresponding 7-demethyl derivatives. An in vitro membrane binding study showed a high affinity (Ki values) of the two ligands for A2A receptor: 8.9 nM for IS-DMPX and 7.7 nM for BS DMPX, and a high A2A/A1 selectivity: > 1100 for IS-DMPX and 300 for BS-DMPX. In mice, [11C]IS-DMPX and [11C]BS-DMPX were taken up slightly more in the striatum than in the reference regions such as the cortex and cerebellum. The uptake ratios of striatum to cortex and striatum to cerebellum gradually increased but were very small: 1.6-1.7 for the striatum-to-cortex ratio and 1.2 for the striatum-to-cerebellum ratio at 60 min postinjection. The uptake by these three regions was reduced by co-injection of an excess amount of carrier or an A2A antagonist KF17837, but not by an A1 antagonist KF15372. The blocking effects in the three regions were greater for [11C]BS-DMPX (32-57%) than for [11C]IS-DMPX (6 29%). Ex vivo autoradiography confirmed that the two ligands were slightly concentrated in the striatum. [11C]BS-DMPX showed more selective affinity for adenosine A2A receptors than [11C]IS-DMPX, but these results have shown that the two tracers were not suitable as in vivo ligands because of low selectivity for the striatal A2A receptors and a high nonspecific binding. PMID- 11023025 TI - Relationship between cancer cell proliferation and thallium-201 uptake in lung cancer. AB - Although thallium-201 (201Tl) uptake is related to perfusion in many normal tissues, the biologic rationale for 201Tl uptake in tumors is uncertain. To determine if tumor uptake is related to cell proliferation, we correlated the relative retention of 201Tl in lung tumors with expression of Ki-67, an indicator of cell proliferation. METHODS: Sixty patients with lung tumors, included small cell carcinoma (n = 8) and non-small cell carcinoma (n = 52), underwent 201Tl single photon emission computed tomography (SPECT) imaging. The 201Tl lesion uptake was determined on early and delayed images and the radiotracer retention index (RI) was calculated. Tumor specimens were obtained at surgery or bronchoscopy. The cell proliferation ratio was estimated with MIB-1, a monoclonal antibody that recognized the nuclear antigen Ki-67. RESULTS: The average 201Tl index was 2.13+/-0.61 (early) and 2.46+/-0.83 (delayed). The average RI was 17.44+/-35.01. Overall, the 201Tl index (delayed) and the cancer cell proliferation were correlated (r = 0.70, p < 0.0001). Of interest, there was a significant correlation (r = 0.872, p < 0.0005) between the 201Tl index on delayed images and the cell proliferation ratio in patients with small cell but not non-small cell lung carcinoma. The 201Tl index (delayed) was significantly higher (p < 0.0001) in patients with small cell lung carcinoma than in patients with non-small cell lung carcinoma. CONCLUSION: 201Tl imaging appears to be useful for evaluating patients with small cell lung carcinoma but not non-small lung carcinoma, and is correlated with the monoclonal antibody MIB-1, a marker of cell proliferation. PMID- 11023026 TI - Clinical evaluation of 99mTc-Technegas SPECT in thoracoscopic lung volume reduction surgery in patients with pulmonary emphysema. AB - 99mTc-Technegas (Tcgas) SPECT is useful for evaluating the patency of the airway and highly sensitive in detecting regional pulmonary function in pulmonary emphysema. The aim of this study is to evaluate regional ventilation impairment by this method pre and post thoracoscopic lung volume reduction surgery (LVRS) in patients with pulmonary emphysema. METHODS: There were 11 patients with pulmonary emphysema. The mean age of patients was 64.1 years. All patients were males. LVRS was performed bilaterally in 8 patients and unilaterally in 3 patients. Post inhalation of Tcgas in the sitting position, the subjects were placed in the supine position and SPECT was performed. Distribution of Tcgas on axial images was classified into 4 types, A: homogeneous, B: inhomogeneous, C: hot spot, D: defect. Three slices of axial SPECT images, the upper, middle and lower fields were selected, and changes in deposition patterns post LVRS were scored (Tcgas score). RESULTS: Post LVRS, dyspnea on exertion and pulmonary function tests were improved. Pre LVRS, inhomogeneous distribution, hot spots and defects were observed in all patients. Post LVRS, improvement in distribution was obtained not only in the surgical field and other fields, but also in the contralateral lung of unilaterally operated patients. In 5 patients some fields showed deterioration. The Tcgas score correlated with improvements in FEV1.0, FEV1.0% and %FEV1.0. CONCLUSION: Tcgas SPECT is useful for evaluating changes in regional pulmonary function post LVRS. PMID- 11023027 TI - Phantom study of fusion image of CT and SPECT with body-contour generated from external Compton scatter sources. AB - PURPOSE: A phantom study was conducted to evaluate the feasibility of body contour definition with Compton scatter photons from external sources of technetium-99m pertechnetate (Tc-99m) to create a fusion image of CT and SPECT images. METHODS: External sources of 1 mCi (37 MBq) Tc-99m were placed on each collimator, and body-contour SPECT images were obtained with an energy window of 100 keV +/- 25% for detecting 90 degrees and 180 degrees Compton scatter photons of Tc-99m from the body surface in water-filled cylindrical and hexagonal phantoms, and in a chest phantom with a Tc-99m-avid simulated lung nodule and multimethod surface markers. In the chest phantom, each transaxial SPECT slices was registered with the corresponding CT slice by using image-matching soft ware. A summation of the registered images yielded a three-dimensional (3-D) fusion image of this phantom. RESULTS: This method clearly visualized the body contour on all the SPECT slices in all the phantoms except for the complex hexagonal phantom. There was no significant difference between the known and SPECT-measured diameters of the cylindrical phantom. The fit of CT and SPECT images of the chest phantom was achieved with a mean alignment error of 5% in visual inspection, which was improved to 0.2% after correction of the magnification of the SPECT images according to the resultant dimensional differences. The 3-D fusion image of this phantom effectively visualized the anatomic location of the lung nodule and surface markers. CONCLUSION: This simple method effectively provided boundary information on the cold phantoms. Although further improvements in the registration technique with CT images are desirable, the body-contour SPECT image obtained by this method has the potential for accurately creating a 3-D fusion image with CT images, and is a feasible way of anticipating the anatomical localization of a target tissue. PMID- 11023028 TI - Fundamental study of hot spot detectability in 3-dimensional positron emission tomography. AB - The purpose of this study was to investigate the detectability of small hot lesions with the 3-dimensional transmission/emission (3D T/E) acquisition mode in FDG-PET scan. The correlation of target detectability, target size, target to non target uptake ratio (T/N ratio) and standardized uptake value (SUV) were studied. Small hot lesions ranged from 4.4 mm to 36.9 mm in diameter were located in cylindrical phantom. The images of phantoms with a T/N ratio of 2.0, 4.0, 6.0, 8.0, 9.6, 13.2, 17.5, 23.8 and 30.3 were obtained with 2-dimensional transmission/emission (2D T/E) scan and 3D T/E scans. Targets in diameter more than 10.6 mm in diameter with an actual T/N ratio ranged from 6.0 to 30.3 could be identified on the images obtained with all the 2D T/E and 3D T/E acquisition modes. The detectability efficiency of small hot target in 2D T/E and 3D T/E scans was as same (77.8%). The T/N ratio of targets from 2D T/E images was 30% to 48.4% different to that from 3D T/E image, and the SUV of the target from the 2D T/E images was almost the same as that from 3D T/E images. This study revealed that 3D T/E scanning had similar hot spot detectability to 2D T/E scanning; 3D T/E and 2D T/E scanning had the same faculty for semiquantitative analysis using SUV. These findings may be helpful for the diagnosis and understanding of 3D T/E FDG-PET in hot lesion detection. PMID- 11023029 TI - Preclinical evaluation of [11C]SA4503: radiation dosimetry, in vivo selectivity and PET imaging of sigma1 receptors in the cat brain. AB - Our previous in vivo study with rats has demonstrated that 11C-labeled 1-(3,4 dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine ([11C]SA4503) is a potential radioligand for mapping CNS sigmal receptors by positron emission tomography (PET). In the present study, we further characterized this ligand. The radiation absorbed-dose of [11C]SA4503 in humans estimated with the tissue distribution in mice, was higher in the liver, kidney and pancreas than in other organs studied, but was low enough for clinical use. The brain uptake of [11C]SA4503 in mice was reduced to approximately 60-70% by co-injection of carrier SA4503 and haloperidol, but not by co-injection of any of six ligands for sigma2 or other receptors, for which SA4503 showed in vitro >100 times weaker affinity than for signal receptor. In the cat brain, the uptake in the cortex was higher than that in the cerebellum. The radioactivity in the cortex and cerebellum accumulated for the first 10 min and then gradually decreased until 81.5 min in the baseline measurement, but rapidly decreased in the carrier-loading condition. The receptor mediated uptake was estimated to be approximately 60-65% of the total radioactivity in the cortex and cerebellum at 76 min after tracer injection. We have concluded that [11C]SA4503 has the potential for mapping sigma1 receptor by PET. PMID- 11023030 TI - Uptake of 99mTc-tetrofosmin, 99mTc-MIBI and 201Tl in malignant thymoma. AB - 99mTc-tetrofosmin, Thallium-201-chloride (201Tl) and 99mTc-MIBI imagings were performed in a patient with malignant thymoma. Tracer uptake in the primary tumor was demonstrated. The tumor-to-background ratios of planar and SPECT imagings were 1.60 and 1.98 for 99mTc-tetrofosmin, 1.12 and 2.09 for 201Tl, and 1.19 and 1.80 for 99mTc-MIBI, respectively. In another patient 99mTc-tetrofosmin and 201Tl imagings were performed. Not only the primary tumor but also the direct invasions and metastatic lesions (bone metastases) were clearly detected. The tumor-to background ratios of planar and SPECT imagings were 2.31 and 2.78 for 99mTc tetrofosmin and 2.45 and 3.58 for 201Tl, respectively. In 99mTc-tetrofosmin scintigraphy we acquired delayed images, and the tumor-to-background ratios of planar and SPECT delayed images were 1.20 and 1.86, the retention ratios were 1.11 and -0.92 and the retention indices were -48.1 and -33.1, respectively. Our preliminary results suggest that 99mTc-tetrofosmin is useful in detecting not only the primary tumor but also metastatic lesions from malignant thymoma. PMID- 11023031 TI - Resolution of Ga-67 citrate uptake in the left neck mass of Hodgkin's disease and reversion of double scoliosis of cervical-thoracic and lower lumbar vertebrae. AB - A 6-yr-old boy underwent a total body Ga-67 citrate imaging study because of a large mass of Hodgkin's lymphoma in the left neck and the left anterior chest wall region. The images showed intense uptake in the left neck extending anteroinferiorly to the left upper chest wall corresponding to the left neck and chest region. In addition, there was mild cervical-upper thoracic scoliosis with convexity to the right and mild scoliosis of the lower lumbar scoliosis with concavity to the left. After three cycles of chemotherapy, in the follow-up Ga-67 citrate total body images seven months after his first Ga-67 citrate imaging, the intense uptake in the left neck and the left upper chest wall had been resolved and the scoliosis of the cervical-thoracic and lower lumbar spine had also been reversed to normal. This case shows that a Ga-67 citrate imaging study is useful for first diagnosis and subsequent monitoring of the therapeutic effects in a follow-up imaging. Also Ga-67 citrate imaging provided evidence that the scoliosis had been reversed. PMID- 11023032 TI - Uncommon and dynamic changes detected by 123I-15-(p-iodophenyl)-3-R,S methylpentadecanoic acid myocardial single photon emission computed tomography in a stunned myocardium induced by coronary microvascular spasm. AB - A 55-yr-old man underwent surgery. Soon after the procedure was finished, the patient complained of chest pain, and the electrocardiogram showed increase in the ST-segment in some leads. Emergency angiography showed normal coronary arteries, but there was asynergy in the left ventricle, and delayed filling of contrast medium was observed in the LCA. An intracoronary infusion of isosorbide dinitrate did not improve the delayed filling of contrast medium or ST segment increase in the electrocardiogram. Soon after nicorandil was injected into the LCA, the patient's symptoms, electrocardiogram, and delayed filling of contrast medium dramatically improved. On the second day, initial imaging by 123I-BMIPP myocardial SPECT showed a moderate increase in tracer uptake in the apico anteroseptal region and a moderate decrease in tracer uptake in the lateral region, in which the first left ventriculography showed akinesis, and delayed imaging revealed a moderate increase in tracer uptake in the apical region and a high washout of 123I-BMIPP in the anteroseptal and lateral regions. On the sixth day, initial imaging by 123I-BMIPP myocardial SPECT showed a moderate decrease in tracer uptake in the apical and lateral regions and a mild decrease in tracer uptake in the anteroseptal region, and delayed imaging revealed a moderate increase in tracer uptake in the apical region and a high washout of 123I-BMIPP in the anteroseptal and lateral regions. By the 30th day, 123I-BMIPP myocardial SPECT had normalized. We consider that these dynamic changes in 123I-BMIPP myocardial SPECT imaging may reflect metabolic changes in fatty acids in the ischemic state, the size of the triacylglycerol pool, and the degree of turnover in the triacylglycerol pool. PMID- 11023033 TI - FDG PET in the clinical evaluation of sarcoidosis with bone lesions. AB - We report a case of sarcoidosis with osseous involvement. Whole-body fluorine-18 fluorodeoxyglucose (FDG) PET scanning demonstrated multiple sites of FDG uptake in the hilar lymph nodes, right ulna, bilateral feet, and subcutaneous tissue of the extremities. FDG PET scanning provides useful information for the detection of sarcoidosis with systemic distribution, even when osseous lesions are included. PMID- 11023034 TI - Diffuse and marked breast uptake of both 123I-BMIPP and 99mTc-TF by myocardial scintigraphy. AB - Unexpected breast uptake was observed in a 32-year-old woman referred for evaluation of hypertrophic cardiomyopathy. Diffuse and marked bilateral breast uptake of 123I-BMIPP and 99mTc-TF was shown by both planar and SPECT imaging during the first study, and the uptake of both radionuclides had decreased significantly eleven months later. At the time of the first radionuclide examination, she was occasionally breast feeding her 2-year-old child and had small amounts of milk production. At the follow up examination, the frequency of breast feeding was significantly reduced and she produced only small amounts of milk. Therefore, the uptake of 123I-BMIPP and 99mTc-TF may have been caused by lactation. PMID- 11023035 TI - In-111 platelets used in evaluation of emboli and thrombi in patients with cerebrovascular accident. AB - Studies with In-111 platelets were conducted to evaluate pulmonary embolus, deep vein thrombus and cardiac thrombus. This study aimed to evaluate active thrombi and possible new emboli in patients with cerebrovascular accident (CVA) in the first 24 hours by using autologous In-111 platelets. Twenty-five patients were included in the study. Carotid artery thrombi observed in 10 patients with this technique were confirmed by Doppler ultrasonography. Intracranial thrombi appearing in 3 cases were verified by X-ray computed tomography (CT). Scintigraphy of 8 patients who showed findings suggesting CVA in CT revealed no abnormal accumulation. This was attributed to the possibility that they were small in size, deep in location and/or were also quite aged. Abnormal accumulations observed in the lungs of 3 patients and in the mediastinum and pelvis in one patient were verified by other radiological methods. In-111 platelet study was found to be useful in patients with CVA to evaluate the active thrombi and possible emboli in the early period before clinical symptoms appeared. PMID- 11023036 TI - Recent advances in bilirubin metabolism research: the molecular mechanism of hepatocyte bilirubin transport and its clinical relevance. AB - Bilirubin is taken up from blood into hepatocytes by sinosuidal membrane transporters and then excreted into bile through the bile canalicular membrane mainly as bilirubin glucuronides. (1) Mechanism of bilirubin uptake into hepatocytes: many organic anions are incorporated into hepatocytes by organic anion transporting polypeptides (rat, oatp1, oatp2, oatp3; human, OATP), liver specific transporter (rlst/HLST), and/or by organic anion transporters (OAT2, OAT3). Oatp1 and HLST transport bilirubin monoglucuronide. However, a transporter of unconjugated bilirubin in the sinusoidal membrane has not as yet been identified. Unconjugated bilirubin may also go across the hepatocyte sinusoidal membrane by a diffusion process. (2) Intrahepatic transport and conjugation of bilirubin: ligandin carries bilirubin to the endoplasmic reticulum (ER) of hepatocytes. In the ER, bilirubin is conjugated by bilirubin uridine diphosphate (UDP)-glycosyltransferase (bilirubin UGT; UGT1A1) to form mono- and diglucuronides of bilirubin. (3) Transport mechanism of bilirubin glucuronides across the hepatocyte canalicular membrane: at the canalicular membrane, bilirubin glucuronides are excreted into bile by multidrug resistance-associated protein 2 (MRP2), a member of the ATP-binding cassette transporter family. (4) Regurgitation of bilirubin glucuronides into blood: MRP3, which is located in the lateral membrane, transports bilirubin glucuronides into blood under conditions of impaired biliary bilirubin excretion. PMID- 11023038 TI - Geranylgeranylacetone stimulates mucin synthesis in cultured guinea pig gastric pit cells by inducing a neuronal nitric oxide synthase. AB - Nitric oxide (NO) has been considered to play an important role in the regulation of blood flow, mucosal integrity, and mucus production in the stomach. We investigated the stimulatory actions of epidermal growth factor (EGF) and a cytoprotective compound, geranylgeranylacetone (GGA), on mucin synthesis in guinea pig gastric pre-pit cells, maintained in a serum-free culture system. GGA increased [3H]glucosamine uptake and the accumulation of mucus granules positive for galactose oxidase-Schiff reaction in the cells. This stimulatory action of GGA was equivalent to that of EGF, but GGA did not stimulate the cell growth. Both EGF and GGA increased the release of NO degeneration products, NO2- and NO3 . The [3H]glucosamine uptake was completely inhibited by the non-selective NO synthase (NOS) inhibitors, N(G)-nitro-L-arginine and N(G)-monomethyl-L-arginine, and it was only partially inhibited by a more selective inhibitor for inducible NOS isoform (iNOS), aminoguanidine. Northern blotting with a cDNA probe for rat iNOS, and Western blotting with a polyclonal antibody against iNOS, demonstrated that GGA did not up-regulate the iNOS mRNA expression nor induce its protein. In contrast, GGA and EGF induced neuronal NOS, but not endothelial NOS, which was confirmed by immunoblot analyses with antibodies against these constitutive NOS isoforms. Thus, the present experiments suggests that GGA, as well as EGF, stimulates mucin synthesis at least in part through an NO-dependent pathway, leading to an increase in the integrity of the gastric mucosa. PMID- 11023037 TI - Hepatic stellate cells: a target for the treatment of liver fibrosis. AB - Hepatic fibrosis is a wound-healing process that occurs when the liver is injured chronically. Hepatic stellate cells (HSC) are responsible for the excess production of extracellular matrix (ECM) components. The activation of HSC, a key issue in the pathogenesis of hepatic fibrosis, is mediated by various cytokines and reactive oxygen species released from the damaged hepatocytes and activated Kupffer cells. Therefore, inhibition of HSC activation and its related subsequent events, such as increased production of ECM components and enhanced proliferation, are crucial goals for intervention in the hepatic fibrogenesis cascade. This is especially true when the etiology is unknown or there is no established therapy for the cause of the chronic injury. This review explores the rationale for choosing HSC as a target for the pharmacological, molecular, and other novel therapeutics for hepatic fibrosis. One focus of this review is the inhibition of two cytokines, transforming growth factor-beta and platelet-derived growth factor, which are important in hepatic fibrogenesis. A number of new agents, such as Chinese herbal recipes and herbal extracts, silymarin, S-adenosyl L-methionine, polyenylphosphatidylcholine, and pentoxifylline are also discussed. PMID- 11023039 TI - Anti-cathepsin G antibodies in the sera of patients with ulcerative colitis. AB - The presence of perinuclear anti-neutrophil cytoplasmic antibodies (P-ANCAs) and that of antibodies against cathepsin G, a target antigen for P-ANCAs, was determined in the sera of patients with ulcerative colitis (UC), relative to the endoscopic severity and disease activity. P-ANCAs were detected by indirect immunofluorescent assay (IIF) on ethanol-fixed human neutrophils. Antibodies to cathepsin G were detected by an enzyme-linked immunosorbent assay (ELISA) and Western blotting. P-ANCAs were detected by IIF in 62.5% of 32 patients with active UC. Anti-cathepsin G antibodies were detected in 40.6% of 32 patients with active UC, and their prevalence was significantly higher in patients with severe colitis, as determined by endoscopy, than in those with mild or moderate colitis (P < 0.05). The prevalence and titers of anti-cathepsin G antibodies were significantly higher during the active than the inactive phase of the disease (P < 0.05). Measurement of titers of anti-cathepsin G antibodies by ELISA in the serum is useful for evaluating the activity of UC. PMID- 11023040 TI - Sex differences in relation to serum hepatitis B e antigen and alanine aminotransferase levels among asymptomatic hepatitis B surface antigen carriers. AB - This study aimed to investigate sex differences in relation to hepatitis B e antigen (HBeAg) and serum alanine aminotransferase (ALT) levels in chronic asymptomatic hepatitis B virus (HBV) infection. HBeAg and ALT level were determined in 636 asymptomatic hepatitis B surface antigen carriers. There was no significant sex differences in the age-adjusted prevalence of HBeAg. Abnormal ALT level (>45 IU/l) was more frequent in carriers with HBeAg (17.5% vs 7.6%; P = 0.001). Multivariate analysis indicated that male sex (odds ratio, 2.0; 95% confidence interval, 1.1-3.6) and HBeAg (odds ratio, 2.6; 95% confidence interval, 1.6-4.3) were independent risk factors for abnormal ALT levels. Male sex and HBeAg-positivity are independent risk factors for abnormal ALT activity in chronic HBV infection. This observation may be related to sex differences in chronic HBV infection. PMID- 11023041 TI - Increased expression of osteopontin in activated Kupffer cells and hepatic macrophages during macrophage migration in Propionibacterium acnes-treated rat liver. AB - Osteopontin is an extracellular matrix component that can act as a chemokine to induce macrophage migration. The significance of osteopontin in macrophage infiltration into the liver was examined in rats given heat-killed Propionibacterium acnes. In normal rats, osteopontin mRNA expression in the liver was minimal, determined by quantitative-competitive reverse transcription polymerase chain reaction (RT-PCR) assay. Northern blot analysis revealed that osteopontin mRNA was not expressed in Kupffer cells isolated from normal rats. When rats received heat-killed P. acnes intravenously, marked macrophage accumulation, forming granulomas, developed in the liver later than 3 days after the injection and its extent became maximal between 5 and 7 days. In these rats, osteopontin mRNA expression was increased in the liver later than 1 day (with its peak at 3 days after the injection), and the mRNA expression was increased markedly in Kupffer cells and hepatic macrophages isolated at 7 days. The mRNA expression of monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-1alpha (MIP-1alpha), chemokines for monocytes and macrophages, was also increased in the liver of P. acnes-treated rats, with peak expression at 3 days. We conclude that osteopontin derived from Kupffer cells and hepatic macrophages may contribute to the infiltration of monocytes and macrophages into the liver cooperatively with the actions of MCP-1 and MIP-1alpha in P. acnes-treated rats. PMID- 11023042 TI - Esophageal intramural pseudodiverticulosis associated with esophageal perforation. AB - We report a rare case of esophageal intramural pseudodiverticulosis with lower esophageal stricture which perforated into the peritoneal cavity after the patient vomited. A 61-year-old man was admitted with severe chest and epigastric pain after dysphagia and vomiting. Under a diagnosis of upper gastrointestinal perforation, laparotomy was performed. The anterior wall of the abdominal esophagus was found to have ruptured, and proximal gastrectomy with abdominal esophagectomy was performed. Histological examination revealed esophageal intramural pseudodiverticulosis with esophageal stricture distal to the site of rupture, and postoperative endoscopy showed diffuse pseudodiverticulosis in the remaining esophagus. The patient is free of symptoms 5 years after the surgery. This case suggests that careful treatment may be indicated in patients with esophageal intramural pseudodiverticulosis with stricture and elevated intraluminal pressure, to minimize the possibility of severe complications such as esophageal perforation. PMID- 11023043 TI - Cronkhite-Canada syndrome: report of two cases. AB - Two cases of Cronkhite-Canada syndrome are reported. In the first case, a 56-year old woman had an adenoma of the colon, arising within the Cronkhite-Canada polyps, which was removed by endoscopic polypectomy. This suggests possible neoplastic transformation of polyps in this syndrome. She achieved remission with corticosteroids, but the polyposis recurred (only in the stomach) 7 months after the remission. In the recurrent polyposis, corticosteroid therapy resulted again in complete remission, which has lasted for 5 years. In the second case, a 69 year-old man developed typical manifestations of the syndrome while under emotional stress. He had a past history of chronic pityriasis lichenoides, and serum antinuclear antibody was positive. These findings suggested a possible role of autoimmune response in the pathogenesis of the syndrome. Corticosteroids were also effective in this patient. PMID- 11023044 TI - Intra-abdominal desmoplastic small round cell tumor (IDSRT). AB - Intra-abdominal desmoplastic small round cell tumor (IDSRT) is a rare neoplasm that develops in the abdominal cavity in young people. We experienced a 27-year old man who visited with ascites of unknown cause. Compression of the colon was found by barium enema examination. On colonoscopic examination, diffuse white elevated lesions, about 5 mm in diameter, surrounded by rubedo, so-called aphthoid lesions, were also observed. IDSRT was diagnosed by biopsy at laparotomy, and chemotherapy with cyclophosphamide, etoposide, doxorubicin, and cisplatin was performed. The tumors shrank temporarily (partial response), but subsequently grew in size again. The patient died during the second course of chemotherapy after relapse. We present one case report, together with a review of the literature. PMID- 11023045 TI - Metastatic gastrinoma to the liver 20 years after primary resection. AB - Gastrinoma is a rare endocrine tumor that is frequently associated with liver metastasis. The liver metastasis is usually seen simultaneously or soon after a primary operation. A 47-year-old woman who had had a total gastrectomy 20 years earlier developed liver metastasis. An interval of this length between surgery and metastasis is extremely rare. The total gastrectomy prevented the patient from developing the usual symptoms of hypergastrinemia that would have enabled early diagnosis of the metastasis. Laboratory examinations on admission revealed a high serum gastrin concentration (1500 pg/ml). Computed tomography showed an irregularly enhanced mass lesion with an uneven, low-density central area in the right anterior inferior segment of the liver. An extended right hepatectomy was performed. Intraoperative ultrasonography showed no abnormalities in the remnant pancreas. Examination of the cut surface of the specimen revealed a yellow, firm, elastic tumor, 55 mm in diameter. The interior of the tumor appeared necrotic. Histopathologically, the tumor was composed of cells with hyperchromatic, dysplastic nuclei arranged in a trabecular pattern with nest formation. Gastrin staining was positive. A histologic diagnosis of metastatic gastrinoma was made. The patient's gastrin concentration returned to normal and she was well at 2-year follow-up. PMID- 11023046 TI - ANCA in inflammatory bowel disease. PMID- 11023047 TI - Chemotactic cytokines (chemokines) in human hepatitis and experimental hepatitis models: which ones play the crucial role? PMID- 11023048 TI - Gnathostomiasis possibly caused by Gnathostoma malaysiae. AB - Gnathostomiasis is rarely reported in travelers, although the disease remains a major public health problem in Southeast Asia. A creeping eruption and Quincke's edema (slowly migrating erythema with pruritus) appeared in two Japanese men who had eaten raw freshwater shrimp in Myanmar. A Gnathostoma larva was found in subcutaneous tissue from one of the men. Four species causing human gnathostomiasis, G. hispidum, G. doloresi, G. nipponicum and G. spinigerum, can be distinguished based on the number of nuclei in intestinal epithelial cells of infected larvae, in cross-section. In G. hispidum, only a single large nucleus is found. Morphologically, our larva was initially identified as G. hispidum. However, since the number of epithelial cells was greater and the body width was larger than those of a "large-type" 3rd-stage larva of G. hispidum, the larva was then identified as a 3rd-stage larva of G. malaysiae, Miyazaki and Dun, 1965, as reported by Setasuban et al, (1991). Since no human cases caused by this species of Gnathostoma have previously been encountered, this appears to be the first report of gnathostomiasis due to G. malaysiae. PMID- 11023049 TI - Histopathological studies of subcutaneous implantation of chlormadinone acetate (CMA) for preventing estrus in queens. AB - The efficacy and clinical safety of chlormadinone acetate (CMA) in preventing estrus were assessed in queens on condition that CMA was subcutaneously implanted in siliastic rubber. Thirteen queens were divided into the following four groups according to dose of CMA-administered: group 1 (n = 3), control; group 2 (n = 3), 2.5 mg/kg; group 3 (n = 3), 5 mg/kg; group 4 (n = 4), 20 mg/kg. The implants were left in these queens for 12 months after implantation. All control animals showed signs of estrus during the experiment, with periods of anestrus of normal duration. In contrast, estrus was completely inhibited in the CMA-treated groups. Histopathologically, the uterus from group 4 had coiled branched glands with little secretion in the endometrium. Mammary glands from the CMA-treated groups showed mild lobular development with acinar proliferation and secretion. Sections through the other organs (pituitary gland, adrenal gland, ovary, and implant site) had no distinct or consistent changes that could be related to the CMA treated. It was concluded, therefore, subcutaneous implantation of CMA could be the good drug-delivery system for reducing changes due to the antigonadotropic and glucocorticoid-like activities and serious condition in the uterine and mammary gland due to progestagenic activity. PMID- 11023050 TI - Effect of antiandrogen, chlormadinone acetate (CMA), in canine spontaneous benign prostatic hyperplasia (BPH). AB - The effect of synthetic steroidal antiandrogen, chlormadinone acetate (CMA), on spontaneous benign prostatic hyperplasia (BPH) in dogs was investigated. Male beagle dogs (5-8 years old) were divided into three experimental groups. Group 1 consisted of untreated controls. Groups 2 and 3 received CMA 0.03, and 0.1 mg/kg/day, p.o., respectively, for 6 months. In group 1, glandular hyperplasia of the prostate was clearly detected. In groups 2 and 3, CMA produced marked atrophy of the glandular epithelium. In addition, a histopathological study showed that CMA medication for 6 months exerted no effect on the testes and adrenals or on immunoreactive LH- and ACTH- cells of the anterior pituitary glands. Therefore, it is suggested that CMA (0.03 and 0.1 mg/kg) causes regression of spontaneous canine BPH without any histopathological effects on the testes, adrenals or anterior pituitary LH- and ACTH-cells. PMID- 11023051 TI - Silastic interposition arthroplasty for osteoarthrosis of the carpometacarpal joint of the thumb. AB - We performed Ashworth's (1977) silastic interposition arthroplasty for painful osteoarthrosis of the carpometacarpal (CM) joint of the thumb. Recently, however, some unsuccessful cases treated by this method have been reported. We therefore reviewed the results of patients treated by Ashworth's method. We reviewed 16 patients operated on according to the original Ashworth method for osteoarthrosis of the first CM joint. The cases consisted of 8 males and 8 females, ranging in age from 32 to 76 years (average, 59.6 years). The patients were grouped according to Eaton's classification as follows: 1 patient was stage II, 6 were stage III, and 9 were stage IV. All patients were followed postoperatively for 1 to 12.5 years (average, 4.5 years). Twelve patients did not complain of any pain but 3 patients felt slight pain when moving their thumb. However, the degree of pain was much less than that experienced preoperatively. Only one patient complained of severe pain 2 years after the first operation. Her implant ruptured and required additional surgery. All except one showed either an increase or no change in grip strength. The range of motion (ROM) was full in all patients. Implants showed sinking in 3 patients, a partial rupture of the implant edge in 5 patients and a whole body failure in only 1 implant, roentogenographically. In spite of these troubles, only 3 patients complained of an unpleasant feeling and 2 reported slight pain. Implant trouble did not always cause the symptoms. The clinical results of this method were mostly satisfactory. PMID- 11023052 TI - Reconstruction of the flexor pollicis longus tendon ruptured, but untreated, during infancy. AB - We treated 3 patients who had ruptured the flexor pollicis longus (FPL) tendon during infancy and which had not been repaired. A two-stage surgical procedure, using a silicone rod, was performed to reconstruct the tendon, and favorable thumb flexion was obtained. A favorable outcome was obtained, even if the gliding of the silicone rod had been poor after the first stage procedure. When the scar of the tendon sheath is available, it should be used as a pulley. When the tendon sheath has completely disappeared, it should be reconstructed. At the second stage of surgery, the flexor digitorum superficialis muscle of the injured finger can be used as a motor source when the muscle is conserved in good condition because its distal stump adheres to the bone. If the muscle is not in good condition, transfer of the flexor digitorum superficialis muscle of the ring finger should be performed. PMID- 11023053 TI - Reducing the volume of polyethylene glycol electrolyte lavage solution to less than 2 liters for bowel preparation. AB - Smaller volumes of polyethylene glycol electrolyte lavage solution (PEG), an oral whole-bowel irrigation solution employed in colorectal preparation, were given to patients to decrease the discomfort associated with pretreatment. Comparison was made between groups receiving standard and modified preparations. A total of 68 patients (gastrointestinal surgery: 55; total colonoscopy: 13) were enrolled in the study. The mean volume of PEG used was significantly smaller in the modified than in the standard preparation (1,694 ml vs. 2,735 ml, p<0.01). In addition, the mean PEG administration period was significantly shorter for the modified preparation (183 min vs. 237 min, p<0.05). However, the mean PEG excretion time and the number of bowel movements were not significantly different between the two groups. There was also no significant difference between the two groups in terms of the efficacy, safety, usefulness, or tolerability of the preparations. These results suggest that it is possible to reduce the PEG volume by more than 1 L to alleviate patient discomfort without a significant loss of efficacy. The modified method is useful for preparing the large bowel for either gastrointestinal surgery or total colonoscopy. PMID- 11023055 TI - The Prince Mahidol awards. PMID- 11023054 TI - Is it permissible to omit mediastinal dissection for peripheral non-small-cell lung cancers with tumor diameters less than 1.5 cm? AB - BACKGROUND: Recently the pros and cons of limited surgery for small-sized peripheral non-small-cell lung cancers (PNSCLCs), such as omission of mediastinal dissection, etc., have been vigorously debated. We analyzed whether hilar/mediastinal lymph node metastases were present in 30 small-sized PNSCLCs. MATERIAL AND METHODS: In the nine years from 1990 to 1998, 294 lung cancer patients underwent lobectomy or pneumonectomy combined with hilar/mediastinal dissection in the Tokai University Hospital. Thirty of these patients diagnosed as having cT1N0M0 PNSCLC with tumor diameters of 1.5 cm or less by computed tomography, are evaluated in this article. RESULTS: The 30 PNSCLC patients consisted of 14 males and 16 females with a mean age of 61 +/- 9 years. Twenty six patients (87%) had no hilar nor mediastinal lymph node metastases (pN0), one patient (3%) had a hilar lymph node metastasis (pN1), and three patients (10%) had mediastinal lymph node metastases (pN2). CONCLUSIONS: Mediastinal lymph node metastases were histologically observed in 3 (10%) of 30 PNSCLC patients with tumor diameters of 1.5 cm or less. Our results show that mediastinal dissection is still necessary even for small-sized lung cancers. PMID- 11023056 TI - Seasonal and spatial variation in natural densovirus infection in Anopheles minimus S.L. in Thailand. AB - We report the first detection of a mosquito densovirus in anophelines, An. minimus species A and species C, and describe temporal and spatial variation in natural densovirus infection. A total of 814 (136 species A; 678 species C) adult mosquitos, obtained over a one year period from human biting catches at three locations in a village in western Thailand, were PCR tested for densovirus infection. Overall infection prevalence did not differ between species (15.4% species A; 14.5% species C). Infection prevalence showed significant seasonal variation. Some spatial heterogeneity in infection was also noted, with timing of peak infection prevalence varying between sites. PCR-screening of An. minimus s.l. larvae found an overall infection prevalence of 18.8%. Larval infection showed a significant positive association with rainfall recorded two months previous to larval collection. Infection in adult mosquitos showed a moderate relationship to environmental variables, but a significant negative correlation with larval infection in the previous month. PMID- 11023057 TI - Transfusion requirements in patients with dengue hemorrhagic fever. AB - Dengue viruses are endemic in Thailand and Southeast Asian countries. A retrospective study of 175 patients with dengue virus infection admitted at the Department of Pediatrics, Ramathibodi Hospital in 1997 was carried out. Fifteen and 160 patients were clinically diagnosed with dengue fever and dengue hemorrhagic fever (DHF), respectively. DHF was commonly found in patients whose ages ranged from 10 to 14 years. The mean body weight was at the 54th percentile for age. In the management, 10.6% of patients with DHF required blood component therapy which included platelet concentrate (64.7%) in patients who exhibited active bleeding, packed red cells (47%) in patients who exhibited a rapid drop in the hematocrit and fresh frozen plasma (29.4%) in patients with circulatory failure who did not respond to intravenous fluid. The transfusion requirement was significantly correlated with the occurrence of bleeding (p < 0.008) and bleeding in the gastrointestinal tract (p < 0.0001) but not correlated with the number of platelet counts (p = 0.207). As a result, physicians in charge should be aware of the transfusion requirement and communicate this to the blood bank in advance for the preparation of appropriate blood components. PMID- 11023058 TI - HIV-infected women delivering without antenatal care in a large Bangkok hospital, 1997. AB - The aim was to estimate the proportion of HIV-infected women giving birth at a large Bangkok hospital who had not received antenatal care (ANC) and to identify predictors of not receiving ANC. At Rajavithi Hospital, Bangkok, women with ANC are routinely tested for HIV at their first antenatal visit; women without ANC are routinely tested at delivery. Hospital staff interview all HIV-infected women and record sociodemographic and HIV risk factor information in a delivery room log book. We abstracted and analyzed data recorded in this log book for all HIV infected women who gave birth at Rajavithi Hospital in 1997. Of 303 HIV-infected women who gave birth, 75% had received ANC at Rajavithi Hospital, 10% had received ANC at other locations, and 15% had not received ANC. On multivariate analysis, HIV-infected women who had received ANC were more likely to work or have partners who worked in construction (25% vs 11%; adjusted odds ratio [AOR]; 2.6; p = 0.03) or have a history of injection drug use (4% vs 0.4%; AOR = 20.8; p = 0.02) than those who had not received ANC, but were less likely to report their current partner as a risk factor for acquiring HIV infection (22% vs. 40%; AOR = 0.4; p = 0.05). Because a substantial number of HIV-infected women give birth in this large Bangkok hospital without receiving ANC, interventions are needed to increase the number of HIV-infected women who receive ANC and to prevent perinatal HIV transmission from HIV-infected pregnant women who have not received ANC. PMID- 11023059 TI - Seroprevalence of hepatitis A virus and varicella zoster antibodies in a Javanese community (Yogyakarta, Indonesia). AB - Hepatitis A virus (HAV) cause an acute inflammation of the liver. Varicella zoster virus (VZV) cause chickenpox (varicella) and herpes zoster. Effective vaccines against hepatitis A and varicella are available for children, adolescents and adults. In order to implement an appropriate vaccination policy, a baseline to assess the potential benefits and sections of the population who would benefit most are required. We investigated seroprevalence of hepatitis A virus and varicella zoster antibodies in a Javanese community. A total of 1,103 subjects were studied. The 600 subjects aged 4 to 9 years were sampled between 23 October and 2 November, 1995. The other subjects were sampled between 12 October and 1 November, 1996. The overall prevalence of anti-HAV in cohort was 28.7%. Anti-HAV seroprevalence rates were below 30% until the age of 15 and below 40% until the age of 25. The anti-varicella seroprevalence showed only in two thirds of seropositive population at the age of 15. The results of the study have implications for vaccination strategies for both hepatitis A and varicella zoster. PMID- 11023060 TI - Current seroprevalence of hepatitis A virus infection among kindergarten children and teachers in Taiwan. AB - Taiwan was a hyperendemic area for hepatitis A virus (HAV) infection before the late 1980s. The seroprevalence of HAV infection was higher than 90% with most HAV infection occurring during childhood. This study was to estimate the seroprevalence of HAV infection among preschool children in central Taiwan. A community-based survey was carried out in 54 kindergartens in 10 urban areas, 10 rural areas and 2 aboriginal areas randomly selected through stratified sampling. Serum samples of 2,549 healthy preschool children and 104 teachers in study kindergartens were screened for the HAV antibodies (anti-HAV) by means of a commercially available microparticle enzyme immunoassay (AxSYM HAVAB). Among aboriginal kindergarten children, more than 96% of them were anti-HAV seropositive due to a mass HAV vaccination program. In urban and rural areas, kindergarten children had a very low prevalence of anti-HAV (0.4%) in contrast to a high seroprevalence in their teachers (78%). There was no gender difference in seroprevalence of anti-HAV, while the anti-HAV seroprevalence was significantly higher in urban areas than in rural areas. Crowdedness of living in urban areas might facilitate the person-to-person transmission of infectious agents. PMID- 11023061 TI - Immunogenicity and safety of an inactivated hepatitis A vaccine in Taiwanese adults and children. AB - The safety and immunogenicity of an inactivated hepatitis A vaccine (AVAXIM, 160 antigen units) was evaluated in 190 subjects: 50 children aged from 2 to 5 years, 70 children aged from 6 to 17 years and 70 adults aged from 18 to 30 years in a monocentric, open, non-controlled, phase III trial conducted in Taipei, Taiwan from December 1996 to October 1997. The vaccine was administered intramuscularly, with a two-dose schedule 6 months apart. Clinical adverse events were monitored during the seven days following each injection. Hepatitis A virus (HAV) antibody titers were measured by modified radioimmunoassay on the day of inclusion and four weeks after both the first dose and booster injection. Among the 190 subjects who received the first dose, 174 (91.6%) were initially HAV seronegative and 16 (8.4%) were HAV seropositive at inclusion. One hundred and seventy-four subjects (91.6%) received the booster dose and completed the study. One month after the first dose, all the subjects, whatever the age, presented HAV antibody titers over 20 mIU/ml. In children (2 to 17 years), the GMT was 136 mIU/ml at week 4 and 7,906 mIU/ml four weeks after the booster dose. In adults (> or = 18 years), GMT values were 93 mIU/ml at week 4 and 3,655 mIU/ml four weeks after the booster. These results show a strong anamnestic response to the second dose of vaccine and are compatible with long-term antibody persistence in each age group. The vaccine was safe and well tolerated. No vaccine-related serious adverse event occurred. No immediate reaction occurred. The majority of the reactions were reported by adults after the primary injection. Local reactions (pain and redness) were reported by 9.0% and 4.0% of the subjects after the primary and the booster doses, respectively. Systemic reactions (mainly myalgia/arthralgia or asthenia) affected less than 10% of the subjects after the first dose and less than 3% after the booster. Results from this study in a Taiwanese population are consistent with those obtained with the same vaccine in previous European studies in children and adults, and suggest that AVAXIM (160 AU) is suitable for use in all subjects aged over 2 years. PMID- 11023062 TI - Clinico-epidemiology of hepatitis B viral infection in Northeastern Thailand. AB - Hepatitis B viral (HBV) infection is a common disease world wide. A study of clinico-epidemiology of HBV infection was conducted in 381 patients who seropositive for hepatitis B surface antigen (HBsAg) in Srinagarind Hospital, Khon Kaen University, Northeastern Thailand, during August 1997 to December 1998. 293 males, 88 females and their mean age was 30.96 +/- 12.78 years with a range from 15 to 77 years. The clinical features of acute hepatitis, chronic hepatitis, liver cirrhosis, hepatocellular carcinoma (HCC) and asymptomatic carrier were 2.36, 34.12, 4.99, 1.05 and 57.48% of cases. Possible routes for HBV transmission were family history of hepatitis, tattooing, intravenous drug addict and blood transfusion in 20.3, 11.3, 8.2 and 6.9% of cases, respectively. Signs of chronic liver disease were common in liver cirrhosis and HCC. Acute fulminating hepatitis was not found in this study. PMID- 11023063 TI - Detection of poliovirus, hepatitis A virus and rotavirus from sewage and water samples. AB - A modified adsorption-elution technique for concentration of enteric viruses from sewage and water samples was developed. The viruses in water were concentrated by negatively charged membrane filtration, eluted with 2.9% tryptose phosphate broth containing 6% glycine pH 9.0, and reconcentrated using centrifugation by a speedVac concentrator. The presence of poliovirus, hepatitis A virus (HAV) RNA, and rotavirus antigen was determined by cell culture isolation, nested polymerase chain reaction (nested PCR), and enzyme-linked immunosorbent assay (ELISA), respectively. A total of 100 sewage and water samples were collected from various sources in congested communities in Bangkok, concentrated and examined for those enteric viruses. Of 20 surface water samples from canals which located near sewage drains, 15% were positive for HAV RNA by nested PCR. Of 48 domestic sewage samples from man-holes of underground sewers, 8% were positive for rotavirus antigen by ELISA. Even though the samples were concentrated 256-2,000 fold, poliovirus was not found by isolation in cell culture. PMID- 11023064 TI - Rapid detection of polioviruses in environmental water samples by one-step duplex RT-PCR. AB - This study describes the rapid detection of polioviruses in environmental waters by a simple reverse transcriptase-polymerase chain reaction (RT-PCR) using two primer pairs for differentiation of poliovirus from non-polio enteroviruses in a single reaction by a one-step method, combining RT and PCR in a single tube. The detection by agarose gel electrophoresis yielded 2 bands of 153-bp and 293-bp for poliovirus tested without the need for further hybridization. The detection sensitivity of this one-step duplex RT-PCR, as measured with RNA extracted by heat treatment from supernatant of infected cell extracts, was 10(-1) 50% tissue culture effective doses (TCID50). This assay was used to evaluate the ability of sample concentration by membrane filter-based adsorption and elution, and purification by a simple RNA isolation based on guanidine isothiocyanate-phenol chloroform extraction; the system yielded a detection limit of 5 x 10(-1) TCID50 seeded in 5 liters of tap water. This protocol was applied to the poliovirus detection in environmental water collected from 2 communities in Bangkok, Thailand during February and May 1998. Of 100 samples tested, 2 water samples collected from the same open sewage pipeline at one location were positive for polioviruses and one sample collected from another sewage pipeline was positive for non-polio enterovirus while a further 97 water samples were negative for both polioviruses and non-polio enteroviruses. With poliovirus detection by cell culture technique, none of the 100 samples tested was positive for poliovirus type 1, 2 or 3. RT-PCR was more sensitive, rapid, simple and cost-effective than the cell culture technique since the two water samples which were positive for polioviruses by RT-PCR failed to be detected by cell culture. Sequence data of 293-bp amplicons from positive samples were compared with those of reference poliovirus strains in the Genbank and the EMBL databases and identity to the sequence of type 1 strain Sabin was found to be 99%. PMID- 11023065 TI - Incidence and clinical manifestations of influenza in nurse assistant students. AB - A prospective study was conducted to find the incidence and clinical manifestations of influenza in 201 nurse assistant students of Faculty of Tropical Medicine during June 1998 to May 1999. There were 106 episodes of influenza-like illness (incidence 52.7%) of which only 33% were proven to be influenza (incidence 17.4%). Main clinical manifestations of influenza included headache, fever, malaise, myalgia, rhinorrhea, cough, and sore throat. We found that influenza could not be diagnosed solely by using clinical manifestations. Respiratory pathogenic bacteria were rarely isolated in patients with influenza like illness and this led to our suggestion that routine pharyngeal culture and antibiotic therapy would not be helpful. Influenza vaccination of every nurse assistant student would be beneficial. PMID- 11023066 TI - Preliminary study on potential circulation of arenaviruses in the rodent population of Nakhon Pathom Province, Thailand and their medical importance in an evoluting environment. AB - Preliminary serological investigations were prefered to detect evidence of arenavirus infection in rodents. The study examined virus antibody in 367 rodents trapped in 6 different geographical areas of Nakhon Pathom Province, Thailand from February-March, 1998. The overall seroprevalence among rodents was 13.3%, mostly in Bandicota savilei (35.7%) and Rattus norvegicus (31.5%). Between ecology, behavior and sex of the rodents, seroprevalence was not significantly different (p>0.05), however the seroprevalence found among different geographical areas of Nakhon Pathom Province were significantly different (p<0.0001). PMID- 11023067 TI - p53 status and human papillomavirus infection in Thai women with cervical carcinoma. AB - Loss of p53 function has been implicated in a wide variety of human malignacies. Many studies suggest that in cervical carcinoma p53 function is inactivated either by gene mutation or by complex formation with E6 oncoprotein product of high-risk human papillomavirus (HPV). The aim of this study was to determine the status of HPV infection and p53 gene mutation as well as their correlation in cervical carcinomas. Formalin-fixed paraffin-embedded tissues of 12 cervicitis, 21 cervical intraepithelial neoplasia grade 3 (CIN 3) and 17 squamous cell carcinomas were determined for the presence of HPV using polymerase chain reaction (PCR) amplification and dot blot hybridization. The status of p53 mutations in exons 5-8 was evaluated by polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) and confirmed by direct nucleotide sequencing. HPV infections were detected in all CIN 3 and squamous cell carcinomas (100%). Mutations of p53 were present in 3 of 38 HPV-positive samples: one with an ATG-->TTG transversion (Met-->Leu) in codon 237 of exon 7; and the others with a TGC-->TGG transversion (Cys-->Trp) in codon 242 of exon 7, and a CGT-->CCT transversion (Arg-->Pro) in codon 273 of exon 8, respectively. Our findings show that the frequency of p53 mutation is low in primary cervical carcinoma and that the p53 gene mutation and HPV infection are not mutually exclusive events in the development of cervical cancer. Thus, other genetic events independent of p53 inactivation may also significantly contribute to the carcinogenesis of the uterine cervix. PMID- 11023068 TI - Epidemiological analysis of methicillin resistant Staphylococcus aureus in Thailand. AB - The geographical distribution of 65 clinical isolates of methicillin resistant Staphylococcus aureus (MRSA) recovered from 7 hospitals in Thailand was investigated. The presence of mecA gene in MRSA was determined by specific PCR with the use of primers 5'-GTAGTTGTCGGGTTTGGT-3' and 5'-GGTATCATCTTGTACCCA-3'. Chromosomal DNA restriction analysis with SmaI was resolved by pulsed-field gel electrophoresis (PFGE) compared with antibiotype analysis and phage type analysis. All 65 strains carried mecA gene. They all were resistant to penicillin, tetracycline, erythromycin, amoxicillin/clavulanic acid and variably resistant to gentamicin, ofloxacin, trimethoprim-sulfamethoxazole, chloramphenicol, fosfomycin and clindamycin; and all isolates were susceptible to vancomycin. A total of 19 PFGE patterns designated as type A, A1, A2, A3, A4, B, B1, C, D, E, E1, E2, F, F1, F2, G, H, I and J was identified. Type A4 and E were commonly found in every studied areas. Phage typing showed even greater variability that 52 (80%) isolates belonged to 25 different phage types; 13 (20%) isolates were non-typable. The clarity and polymorphism of the PFGE patterns enable us to discriminate between isolates which could not be differentiated by antibiogram or phage type analysis. The findings demonstrate the existence of a common epidemic MRSA clone in Thailand. PMID- 11023069 TI - Rapid isolation and detection of Escherichia coli O157:H7 by use of rainbow agar O157 and PCR assay. AB - This study has evaluated the use of a commercially available Rainbow agar O157 and polymerase chain reaction (PCR) assays for the detection of Shiga-like toxin producing Escherichia coli and to serotype E. coli O157:H7 from raw meat. The Rainbow agar O157 was found to be selective and sensitive for the screening of the E. coli O157 from artificially and naturally contaminated meat samples. Shiga like toxin producing E. coli were identified with two primer pairs that amplified fragments of the SLT-I (384 bp) and SLT-II (584 bp). E. coli O157:H7 was serotyped with a primer pair specified for the H7 flagellar gene, which amplify specific DNA fragments (625 bp) from all E. coli O157:H7 strains. The use of Rainbow agar O157 described allows for the presumptive isolation of E. coli O157 in 24 hours. Identification and confirmation of the presumptive isolates as E. coli O157:H7 by PCR assays require additional 6-8 hours. The above-mentioned screening and identification procedures should prove to be a very useful method since it allows for the specific detection of E. coli O157:H7. PMID- 11023070 TI - Episome profiles and mobilizable beta-lactamase plasmid in Haemophilus ducreyi. AB - Chancroid caused by Haemophilus ducreyi has been described as a significantly predisposing factor of HIV heterosexual transmission in an endemic region of both diseases. The fastidious, H. ducreyi has been reported world wide with various antimicrobial susceptibility patterns. A high tendency of drug resistances has generally been found among isolates derived in Thailand. In this study, the plasmids of H. ducreyi were isolated and analysed from 63 clinically derived organisms. Twenty-nine out of 63 isolates (46%) revealed the same plasmid profiles. Plasmid DNA was further cloned into Escherichia coli and transformants were selected. A 3.6 kb plasmid (pCb) carrying ampicillin resistance was subsequently identified. The pCb conferred resistance to various beta-lactam antibiotics including penicillin G, carbenicillin, piperacillin, cefazolin, cefoperazone, ampicillin-sulbactam, and amoxicillin-clavulanate but not to cefoxitin. Co-resistance to streptomycin, chloramphenicol and tetracycline was not detected. Beta-lactamase gene was located on the major pCb fragment of EcoRI and AatII cutting. PMID- 11023071 TI - The comparison of tuberculosis treatments: a short course therapy and the directly observed short course treatment (DOTS), East Java Province, Indonesia. AB - Tuberculosis has been given great attention as HIV/AIDS has increased. Because HIV causes a higher tuberculosis risk is becoming more and more important better tuberculosis treatment. This study aimed to compare the smear conversion rate between short course therapy and the tuberculosis treatment with directly observed short course therapy (DOTS), in East Java, Indonesia. The average smear conversion rate in short course therapy among 35,292 cases was 94.40% over 5 year period (1989/90-1993/94). The tuberculosis treatment with DOTS was started in 1994/95. In the first 2 years the smear conversion rate were 97.67% (42/43) and 98.00% (196/200), respectively. The smear conversion rate of the treatment with DOTS was significantly higher compared to a short course therapy (p-value: <0.001). Thus, tuberculosis treatment with DOTS should be promoted. The concept of supervision by health workers or health cadres should be applied considering mostly are given by family members. And there should be readiness of tuberculosis staff to do the treatment with DOTS in all levels to expand the coverage. PMID- 11023072 TI - Imipenem therapy for septicemic melioidosis in a child with penicillin and cephalosporin adverse reaction. AB - The recommended treatment for severe melioidosis is ceftazidime or a combination of ceftazidime and trimethoprim-sulfamethoxazole (TMP/SMX). Amoxicillin clavulanate has been shown to be an effective alternative therapy. In patient who is allergic to penicillin and cephalosporin, imipenem an alternative drug may be used. We described a 10 year-old boy who was diagnosed as septicemic melioidosis and type 1 diabetes mellitus. He developed fever and rash while being given ceftazidime and TMP/SMX. The fever recurred when amoxicillin-clavulanate was administered orally. He was successfully treated with imipenem. PMID- 11023073 TI - Factors associated with TT (tetanus toxoid) immunization among pregnant women, in Saparua, Maluku, Indonesia. AB - A cross sectional study was conducted at Saparua Health Center to determine factors associated with the administration of tetanus toxoid. In all, 64 pregnant women were recruited. The majority of the women were housewives, aged 17 to 30 years old, and having more than one child. They were educated to high school, knew some information on tetanus toxoid, and knew at least one of the tetanus symptoms. Almost all did not know the cause of tetanus. The logistic regression of knowledge on tetanus and TT immunization: mothers who heard of TT were 1.54 more likely to have been immunized than those who did not, while mothers who knew the use of tetanus toxoid were 2.15 times more likely to have been immunized than those who did not, and those who knew at least one of the tetanus symptoms were 1.86 times more likely to have been immunized than those who did not, respectively controlling other variables constant. Furthermore, women who had antenatal care were 30 times more likely to have been immunized than those who did not. Enhancing mothers knowledge on tetanus is important to increase the coverage of tetanus toxoid. Moreover, antenatal care would cause contact with sources of tetanus toxoid and hence increase the chance of having the immunization. At the same time, this decreases tetanus neonatorum. Considering the majority of pregnant women received information on tetanus from healthworkers, the use a variety of media would be advantageous. PMID- 11023074 TI - Development of EIA for detection of Chlamydia trachomatis in genital specimens. AB - A double antibody sandwich enzyme immunoassay (EIA) for chlamydial antigen detection was developed using a monoclonal antibody against lipopolysaccharide (LPS) of Chlamydia trachomatis as a coating antibody. Polyclonal rabbit antiserum against partially purified antigen from elementary body (EB) antibody and horse radish peroxidase conjugated goat anti-rabbit antibody were used as the primary and secondary antibody respectively. The developed EIA could detect protein of partially purified EB at the lowest concentration of 250 ng/ml. The assay was evaluated against the cell culture (CC), DNA hybridization assay (PACE2 system: Gen-Probe, San Diego, CA, USA) and a commercial enzyme immunoassay (kEIA) (Bioquest, NSW, Australia). The sensitivity, specificity, positive and negative predictive values of the developed EIA (dEIA) were 87, 96.2, 80, 97.7 for the specimens from females and 90.9, 90.7, 71.4, 97.5 for the specimens from males repectively. Cross reaction was not found with Escherichia coli, Acinetobacter anitratus, beta-Streptococcus group A, Enterobacter spp, Enterococcus, Lactobacillus spp, Neisseria spp, but it was found with Candida albicans and herpes simplex virus type 1. The developed EIA can be applied successfully for both genders, particularly males. The cost per test is less than those for CC, kEIA and PACE2. PMID- 11023075 TI - Ex post and ex ante willingness to pay (WTP) for the ICT Malaria Pf/Pv test kit in Myanmar. AB - Willingness to pay (WTP) for the ICT Malaria Pf/Pv test kit was assessed by the contingent valuation method using a bidding game approach in two villages in Myanmar. Kankone (KK) village has a rural health center (RHC) and Yae-Aye-Sann (YAS) is serviced by community health worker (CHW). The objectives were to assess WTP for the ICT Malaria Pf/Pv test kit and to determine factors affecting the WTP. In both villages WTP was assessed in two different conditions, ex post and ex ante. The ex post WTP was assessed at an RHC in the KK village and at the residence of a CHW in the YAS village on patients immediately following diagnosis of malaria. The ex ante WTP was assessed by household interviews in both villages on people with a prior history of malaria. Ordinary least squares (OLS) multiple regression analysis was used to analyze factors affecting WTP. The WTP was higher in ex post conditions than ex ante in both villages. WTP was significantly positively associated with the average monthly income of the respondents and severity of illness in both ex post and ex ante conditions (p < 0.001). Distance between the residence of the respondents and the health center was significantly positively associated (p < 0.05) in the ex ante condition in a household survey of YAS village. Traveling time to RHC had a negative relationship with WTP (p < 0.05) in the ex post condition in the RHC survey in KK village. PMID- 11023076 TI - Transmission of intestinal blastocystosis related to the quality of drinking water. AB - A cross-sectional study was performed to evaluate the risk factors of Blastocystis hominis infection in the Thai army population of the 11th Infantry Division, Chachoengsao Province, Thailand. 201 army personnel and their family members were enrolled in this study. Intestinal parasitic infections in this population were assessed by stool examination using simple smear, formalin/ether technique and Kato-thick smear. Approximately one third of the specimens were positive for one or more intestinal parasites. With the prevalence of 21.9%, B. hominis was the most common intestinal parasite found in this population. Our data indicated that blastocystosis in this army population was significantly linked to the quality of drinking water. After being adjusted for potential confounders, consuming neither filtered nor boiled water was independently associated with blastocystosis. PMID- 11023077 TI - In vitro sensitivity of Trichomonas vaginalis to DNA topoisomerase II inhibitors. AB - Vaginal trichomoniasis is a highly prevalent sexually transmitted disease caused by a microaerophilic protozoan Trichomonas vaginalis. The disease is one of the most common sexually transmitted disease and can augment the predisposition of individuals to human immunodeficiency virus (HIV) infection. Although the disease can be treated with metronidazole and related 5-nitroimidazole, cases of trichomonal vaginitis which are refractory to standard treatment seems to be increasing. Clearly, new antitrichomonad agents are needed and DNA topoisomerase II may acts as a new target for antitrichomonad agents. In this study, in vitro sensitivity of T. vaginalis to DNA topoisomerase II was investigated. Axenic culture of local strain of T. vaginalis was performed. Both eukaryotic and prokaryotic DNA topoisomerase II inhibitors such as ellipticine, amsacrine and fluoroquinolones were tested for effectiveness against T. vaginalis in vitro compared to metronidazole. T. vaginalis was sensitive to metronidazole under aerobic conditions. Minimal inhibitory concentrations (MICs) of eukaryotic DNA topoisomerase II inhibitors, ellipticine and amsacrine, were 6.4 mM and 64 mM, respectively. The MICs of prokaryotic DNA topoisomerase II or DNA gyrase inhibitors; ciprofloxacin, ofloxacin and norfloxacin were 64, 960 and 1,280 mM, respectively. Based on the results, among DNA topoisomerase II inhibitors ellipticine was the most effective drug against T. vaginalis in vitro whereas fluoroquinolones did not show high antitrichomonad activity. PMID- 11023078 TI - Seroprevalence of specific total immunoglobulin (Ig), IgG and IgM antibodies to Toxoplasma gondii in blood donors from Loei Province, Northeast Thailand. AB - A total samples from 345 healthy blood donors from Loei Province, Northeast Thailand were examined for anti-Toxoplasma gondii antibodies by ELISA. The seroprevalence of the anti-Toxoplasma gondii total Ig, IgG and IgM antibodies was 4.9%, 4.1% and 4.3%, respectively. Overall seropositive rate was 33 out of 345 individuals (9.6%). Among the seropositve cases, 5 (15.2%), 2 (6.1%) and 13 (39.4%) of the samples were determined by using each type of anti-T. gondii total Ig, IgG and IgM antibodies, respectively. The seropositive sera was also determined by combining of anti-T. gondii antibodies (anti-T. gondii total Ig with IgG and anti-T. gondii total Ig with IgM antibodies). These results were 10 (30.3%) and 2 (6.1%) cases, respectively. Only one (3%) sample had all types of anti-T. gondii antibodies. In addition, the frequency distribution curves of ELISA optical densities of anti-T. gondii total Ig, IgG and IgM antibodies in blood donor presented "unimodal" curves. The negative results were found in the age group that less than 20 years old and more than 51. The highest seropositive results were found in two age groups (21-30 and 31-40 years old), and males were significantly higher than female (p < 0.05). These results demonstrated that when using anti-T. gondii total Ig, IgG and IgM antibodies for determining the seroprevalence, the sensitivity was twice that with the anti-T. gondii, total Ig antibody alone. PMID- 11023079 TI - An epidemiological study on Opisthorchis viverrini infection in Lao villages. AB - The prevalence of liver fluke, Opisthorchis viverrini, infection in rural and urban communities was studied in Khammouane Province in Lao PDR. The infection was the commonest among the villagers examined, showing the positive rates of 52.9% and 55.0% in two rural communities and 60.7% in an urban community, respectively. The infection rate reached up to 20% or more within 4 years after birth and increased with age to a plateau in the age group over 20 years. The highest prevalence rate was over 80% in the age group of 35-54 years. The age related patterns of infection in males and females were almost the same, although there were some sex-related differences by village and by age group. Infection with Opisthorchis viverrini appears to be a serious public health problem strongly associated with the frequent eating habit of raw fish in low land Laos. PMID- 11023080 TI - A survey of infective larvae of Gnathostoma in eels sold in Ho Chi Minh City. AB - To investigate the distribution of Gnathostoma spp in Ho Chi Minh City (HCM city), 1,081 eels were purchased from a local market twice a month from March 1998 to February 1999. Infective larvae of Gnathostoma spp detected from the flesh and liver of eels by the press preparation technique were examined and identified. Three hundred and fifty advanced third-stage larvae were recovered from liver, none from the flesh. The average rate of infection was 0.11; a high rate of infection was found from August to November and a low rate of infection from February to May. The average number of larvae/eel was 2.9; the greatest number of larvae/eel was in January whereas the lowest was in March and April. There was a marked decrease in both prevalence and intensity of infection from February to May, followed by a rise from June. The finding suggests that in HCM city, the infection rate abruptly decreases soon after the end of the rainy season and starts to rise when the rain comes and reaches its peak at the end of the rainy season. All recovered larvae were identified as G. spinigerum. PMID- 11023081 TI - Gnathostoma spinigerum: analysis of protein patterns by two dimensional gel electrophoresis. AB - The protein extracts from male (MS) and female (FS) adults and advanced third stage larvae (LS) of Gnathostoma spinigerum were separated by high resolution two dimensional gel electrophoresis (2-DE). The polypeptide spots, as detected by silver staining, were subsequently identified. The spot patterns of LS, MS and FS were highly complex and consisted of more than 75, 44, 52 prominent spots, respectively. In addition, the stage-specific protein patterns were identified. This 2-DE database should provide an important reference for future biological and biochemical studies of G. spinigerum. PMID- 11023082 TI - Life table analysis of ascariasis in a rural community in Jiangxi Province, China. AB - The prevalence of ascariasis is very high in parts of rural areas in southern China. The high prevalence of ascariasis is one of the most neglected public health problems. A longitudinal investigation on the prevalence of ascariasis in a rural community in China was reported previously. In this article, we re analyze the reported age specific prevalence of ascariasis using a life table method. The results from our analysis may provide a better summary of the impact of the disease on the studied population. PMID- 11023083 TI - Prevalence of enterobiasis and its incidence after blanket chemotherapy in a male orphanage. AB - A prospective observational study was conducted in a male orphanage to find out the prevalence of enterobiasis and its incidence after blanket chemotherapy using mebendazole. We found that the prevalence of enterobiasis was 28.9%. The incidence density of enterobiasis after blanket chemotherapy was 379.82 per 1,000 person-years which was quite high. We suggest that blanket chemotherapy should be repeated at every 6 months interval to control enterobiasis in orphanages. PMID- 11023084 TI - Comparative studies on the efficacy of three anthelminthics on treatment of human strongyloidiasis in Okinawa, Japan. AB - A study was undertaken to compare the efficacy of three drugs in the treatment of uncomplicated strongyloidiasis in Okinawa, Japan. Two hundred and eleven patients with confirmed Strongyloides stercoralis infection were given either ivermectin, 6 mg in a single dose, albendazole, 400 mg/day for 3 days or pyrvinium pamoate, 5 mg/kg/day for 3 days. For each treatment, the same regimen was repeated once 2 weeks later. Efficacy was evaluated at 2 weeks, 6 months and 12 months after the second course of treatment. Each follow-up examination included a parasitological examination of z stool specimens, using the agar-plate culture method. Coprological cure was obtained in 65 of the 67 patients treated with ivermectin (97.0%), in 65 of the 84 patients treated with albendazole (77.4%) and only in 14 of the 60 patients who were given pyrvinium pamoate (23.3%). The cure rates were lower in males and in the patients with concurrent HTLV-1 infection. An epidemiological feature of Strongyloides infection in recent Okinawa might allow workers to evaluate the exact efficacy of the treatment for an extended period, over a year, without the possibility of reinfection from the environment. PMID- 11023085 TI - Short-term effects of branched-chain amino acids on liver function tests in cirrhotic patients. AB - A randomized study was conducted in 29 ambulatory cirrhotic patients to determine the short-term effects of branched-chain amino acids (BCAA) on nutritional status, biochemical liver function tests and caffeine clearance. Each patient received a 4-week period of isonitrogenous and isocaloric regimens, either a standardized diet contained 40 g protein with supplementation of BCAA 150 g daily (group I) or only a standardized diet contained 80 g protein daily (group II). At the end of treatment, only group I showed significant improvements in transaminase levels as well as the caffeine clearance test compared with those of the pre-treatment levels. Nonetheless, significant improvements in nutritional parameters and additional liver function tests were not yet detected. We conclude that the short-term nutritional supplementation of BCAA is well tolerated and leads to improvement in hepatic metabolic capacity assessed by the caffeine clearance test. PMID- 11023086 TI - Prevalence of dyslipidemia in the elderly in rural areas of Thailand. AB - Dyslipidemia is highly prevalent in the urban areas of Thailand but information in the rural area, particularly in the elderly, is limited. The objective of this study was to determine the prevalence of dyslipidemia in the elderly who live in the rural areas of Thailand. Random sampling of the volunteers aged > or = 60 years in 3 districts of Samut Songkhram and Ratchaburi provinces was done. After 12-hour fast, the blood sampling was drawn for the analysis of total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol and low-density lipoprotein (LDL) cholesterol. Eighty men and 123 women, aged 60-87 years old, were included in the study. Mean serum lipid levels of cholesterol, LDL cholesterol, HDL cholesterol and triglycerides were 261.74+/-47.58, 180.35+/ 45.06, 43.72+/-12.06, and 188.38+/-103.84 mg/dl respectively. Women had significantly higher body mass index, cholesterol and LDL cholesterol levels than men. Seventy percent of them had cholesterol > or = 240 mg/dl and LDL cholesterol > or = 160 mg/dl. Twenty-five percent had HDL cholesterol < or = 35 mg/dl. However, LDL/HDL cholesterol ratio > 5 which indicated high risk for coronary heart disease were found in only 34%. In conclusion, prevalence of dyslipidemia was very high in Thai rural elderly. Further surveillance in this population is essential in verifying the impact of dyslipidemia as a risk of cardiovascular disease in Thai elderly people. PMID- 11023087 TI - Prevalence of non-insulin dependent diabetes mellitus in urban areas of eastern Nepal: a hospital based study. AB - Non-insulin dependent diabetes mellitus (NIDDM), which affects millions of people throughout the world, is a widely prevalent chronic debilitating disease that causes short term and long term complications. It is a problem in a developing country like Nepal, where there has been no report of prevalence. Hence this study was undertaken to investigate the prevalence of NIDDM among urban patients attending the outpatient clinic of BP Koirala Institute of Health Sciences (BPKIHS) hospital, and coming from the eastern part of Nepal. A sample of 1,840 subjects was incorporated in the study during a period of one year. WHO diagnostic criteria (1985) were followed to establish the diagnosis of NIDDM. The prevalence of diabetes was 6.3% (1.63% previous and 4.67% new) which is relatively high in comparison to many other countries. The prevalence of NIDDM in females was relatively lower (5.75%) than in males (6.73%). The prevalence showed an increasing trend with increasing age. The high incidence (new cases) of NIDDM in Nepal as found in the study may be due to lack of public awareness regarding the problem and poor medical services in the country. PMID- 11023088 TI - Characteristics of repeat aborters in Vietnam. AB - Two hundred and sixty married women seeking induced abortion service in Hanoi, Vietnam were interviewed to determine the magnitude of repeat induced abortion and explore selected characteristics of the repeat aborters. Seventy-one percent of the sample reported having had at least one previous induced abortion. After adjustment for age and number of living children, poor attitudes toward contraception, low use of modern contraceptives and failure of contraception were shown to be significantly associated with repeat induced abortion. Woman's age, number of living children, contraceptive knowledge and experience and desire for no more children were positively related to repeat induced abortion. Socio demographic characteristics were not related to repeat induced abortion. Improvement of attitudes toward contraception, persuasion to use modern contraception and promotion of contraceptive effectiveness are recommended strategies to prevent repeat induced abortion. PMID- 11023089 TI - Adult immunization--a neglected issue in Southeast Asia. AB - Adult immunization is a neglected and underpublicised issue in Southeast Asia. Vaccine-preventable diseases cause unnecessary morbidity and mortality among adults in the region, while inadequate immunization results in unnecessary costs, including those associated with hospitalization, treatment, and loss of income. Childhood vaccination coverage is high for the EPI diseases of diphtheria, tetanus and pertussis; however, unvaccinated, undervaccinated, and aging adults with waning immunity remain at risk from infection and may benefit from vaccination. Catch-up immunization is advisable for adults seronegative for hepatitis B virus, while immunization against the hepatitis A and varicella viruses may benefit those who remain susceptible. Among older adults, immunization against influenza and pneumococcal infections is likely to be beneficial in reducing morbidity and mortality. Certain vaccinations are also recommended for specific groups, such as rubella for women of child-bearing age, typhoid for those travelling to high-endemicity areas, and several vaccines for high-risk occupational groups such as health care workers. This paper presents an overview of a number of vaccine-preventable diseases which occur in adults, and highlights the importance of immunization to protect those at risk of infection. PMID- 11023090 TI - Hydatid cyst presenting as a breast lump. AB - We report a 33-year-old woman who presented with a breast lump which, on pathological examination, was found to be a hydatid cyst. There was no evidence of any coexistent lesion elsewhere. To our knowledge, this represents the first case of hydatid disease of the breast reported from Nepal. PMID- 11023091 TI - Pleural effusion in childhood falciparum malaria. AB - Pulmonary complication is a rare manifestation of childhood malaria and isolated pleural effusion without pulmonary edema has never been reported in children. We report here an 11-year-old boy who suffered from cerebral malaria and massive right pleural effusion. The patient was treated with intravenous artesunate, albumin, and other supportive treatments. He recovered completely after eight days. The clinical and laboratory courses suggested that the plasma leakage played a role in the pathogenesis of pleural effusion. PMID- 11023092 TI - Concomitant dengue hemorrhagic fever with Kawasaki disease. AB - This is an unique case report of concomitant dengue hemorrhagic fever and Kawasaki disease, not reported previously. The unusual clinical course of persistent fever, coronary artery involvement evidenced from 2-D echocardiogram, and hydrops of the gallbladder is discussed. PMID- 11023093 TI - Polymorphous low-grade adenocarcinoma: a study of 40 cases with long-term follow up and an evaluation of the importance of papillary areas. AB - Forty cases of polymorphous low-grade adenocarcinoma with a minimum of 10 years of follow up were reviewed. The patients included 13 men and 27 women age 22 to 71 years (median age, 54 years); 30 were white and 10 were black. The tumors were all intraoral, and the palate was the most common site (n = 24). Histologically, the neoplasms were characterized by nonencapsulated, infiltrative borders: bland, regular nuclei; and highly variable growth patterns, including tubular, solid, papillary, microcystic, cribriform (with true lumens), pseudoadenoid cystic (without true lumens), fascicular, single file, and strand-like. Papillary areas of more than focal extent were present in 17 cases, but these cases were otherwise similar to the remainder and were considered to form part of the spectrum of polymorphous low-grade adenocarcinoma. Thirteen patients had local recurrence, which was not controlled by subsequent treatment in six; six patients had cervical lymph node metastasis; three patients had distant metastasis; and five patients died of or with tumor after prolonged periods. There was a statistically significant relationship between more than focal papillary growth and cervical lymph node metastasis, and between positive or unknown surgical margins and local recurrence (although not uncontrolled local recurrence); however, these were the only independent statistically significant correlations found between any clinical or pathologic parameter and any aspect of tumor behavior or patient survival. PMID- 11023094 TI - Renal cell carcinoma with rhabdoid features. AB - Neoplasms with rhabdoid features have been reported at many anatomic sites. In the kidney, rhabdoid tumors are typically found in children, whereas only rare examples have been reported in adults. Little is known of renal cell carcinomas (RCCs) that exhibit rhabdoid features. The objective of this study was to determine the incidence of RCC with rhabdoid attributes and characterize the histologic, immunophenotypic, and ultrastructural features by retrospective analysis of 480 consecutively identified cases of RCC in radical nephrectomy specimens. Immunohistochemical evaluation was performed in cases with rhabdoid foci using a panel of antibodies to pancytokeratin (pan-CK), CK7, CK20, epithelial membrane antigen (EMA), S-100 protein, desmin, vimentin, neuron specific enolase (NSE), muscle-specific actin (MSA), smooth muscle actin (SMA), human melanoma, black-45 (HMB-45), and glial fibrillary acidic protein (GFAP). Electron microscopy was also performed in selected cases. The presence and extent of rhabdoid foci in relation to pathologic stage and grade were assessed. Twenty three of 480 cases of RCC (4.7%) exhibited rhabdoid features. The 23 patients were all adults with a mean age of 61.8 years (age range, 33-84 yrs). Fifteen of the patients were men and eight were women. Histologically, the rhabdoid foci were typified by sheets and clusters of variably cohesive, large epithelioid cells with vesicular and often eccentric nuclei, prominent nucleoli, and large, paranuclear intracytoplasmic hyaline globules (inclusions). The presence of these rhabdoid features was related to high histologic Fuhrman grade of the nonrhabdoid carcinoma component, with an incidence of 0 of 84 grade I cases, eight of 300 grade 2 cases (2.6%), six of 70 grade 3 cases (8.9%), and nine of 26 grade 4 cases (34.6%; p = 3 x 10(-9)). The rhabdoid foci were all high grade. The presence of rhabdoid foci was also found in higher stage carcinomas. A total of 52% (12 of 23) of RCC cases with rhabdoid features exhibited extrarenal extension compared with 28% (24 of 92) of contemporary RCCs without rhabdoid features (p = 0.03). The size of the rhabdoid component ranged from 1 mm to more than 2 cm and comprised 1% to 50% of the renal mass. Immunoreactivity for vimentin (100%), NSE (79%), and panCK (56%) was present in the majority of cases. Substantial percentages of cases were immunopositive for EMA (47%) and S-100 protein (37%), with minimal to no immunohistochemical reactivity for CK7 (5%), SMA (5%), CK20 (0%), desmin (0%), MSA (0%), HMB-45 (0%), and GFAP (O%). A distinctive globular, paranuclear reaction pattern was found for the cytokeratin, EMA, and vimentin immunostains. Ultrastructurally, the rhabdoid cells had paranuclear intermediate filament aggregates or paranuclear condensation of organelles, often associated with peripheral vacuolization. Adult RCCs may harbor a rhabdoid component, and these neoplasms can be regarded as "composite" tumors. Rhabdoid elements are important to identify because of their high-grade nature, and association with high stage. Adult RCC with rhabdoid elements should be distinguished from pure rhabdoid tumors of kidney, in light of their clinicopathologic differences. Rhabdoid differentiation in adult renal cell carcinoma may represent clonal divergence and/ or evolution, and emergence of a particularly aggressive element. PMID- 11023095 TI - Gastrointestinal stromal tumors and leiomyosarcomas in the colon: a clinicopathologic, immunohistochemical, and molecular genetic study of 44 cases. AB - Gastrointestinal stromal tumors (GISTs), mesenchymal tumors largely specific for the gastrointestinal tract, have been well defined in the stomach and small intestine, but have not been extensively documented or contrasted with true smooth muscle tumors in the colon. This study was undertaken to determine the clinicopathologic features of GISTs of the colon, excluding the rectum, and to compare them with leiomyosarcomas (LMSs) of the same location. A total of 37 colonic GISTs and seven LMSs from the files of the Armed Forces Institute of Pathology and the Haartman Institute of the University of Helsinki were analyzed. The GISTs occurred predominantly in adults older than 50 years of age (median, 67 yrs), and most were histologically malignant; four small benign tumors (< or = 1 cm) were incidentally detected, and 10 others had minimal mitotic activity (five or fewer mitoses per 50 high-power fields). The colonic GISTs were typically transmural tumors with frequent intraluminal and outward bulging components. Histologically, they usually showed a spindle cell pattern (92%), whereas 8% were epithelioid. Most tumors (19 of 25) were positive for CD117 (KIT) and for CD34 (16 of 27); six tumors coexpressed alpha-smooth muscle actin and CD117; none showed desmin or S-100 protein. C-kit mutations in exon 11 were seen in 5 (36%) of 14 colonic GISTs. None of the patients with incidental small tumors had a recurrence, whereas 2 of 10 patients with tumors larger than 1 cm but minimal mitotic activity died of the disease with liver metastasis. Nearly all patients whose tumor was larger than 1 cm and showed more than five mitoses per 50 high power fields died of disease; half had evidence of metastasis. LMSs were typically intraluminally bulging, polypoid masses that showed a histologic likeness to differentiated smooth muscle cells. They occurred in five men and two women with a median age of 61 years. Most LMSs were high-grade histologically and showed smooth muscle actin, desmin, or both. All were negative for CD34 and CD117 and lacked c-kit mutations. Five of the seven patients died of disease, and two had a long-term survival, despite high mitotic activity. These results show that KIT-positive GISTs are more common than LMSs of the colon, and these tumor groups have clinicopathologic differences that warrant their separation. PMID- 11023096 TI - Low-grade fibromyxoid sarcoma and hyalinizing spindle cell tumor with giant rosettes: a clinicopathologic study of 73 cases supporting their identity and assessing the impact of high-grade areas. AB - Low-grade fibromyxoid sarcoma (LGFMS) is a rare sarcoma characterized by bland histologic features and a paradoxically aggressive clinical course. The hyalinizing spindle cell tumor with giant rosettes (HSCT) is a closely related tumor characterized by the presence of giant collagen rosettes. Only a single example of a metastasizing HSCT has been reported. A small subset of both LGFMS and HSCT display areas of increased cellularity and atypia which qualify as intermediate- to high-grade sarcoma; the significance of these features has not been definitively assessed. We present the clinicopathologic features of 77 cases of LGFMS and HSCT to determine the degree of overlap of these two lesions, their biologic behavior, and the significance of the occasional presence of intermediate- to high-grade sarcoma within both. The patients (33 female, 40 male) ranged from 3 to 78 years of age (median, 34 yrs). Fourteen cases occurred in patients less than 18 years of age. The tumors measured from 1 to 23 cm (median, 4.5 cm) and occurred predominantly in the trunk and lower extremities in both the deep (66 cases) and superficial (7 cases) soft tissues. In 15 cases, the tumor was present > 1 year before diagnosis. All tumors showed predominantly the typical hypocellularity and bland cytologic features of typical LGFMS; however, areas of hypercellularity and nuclear enlargement and hyperchromatism were identified in 12 of 73 (16%) and 7 of 73 (10%), respectively. Necrosis and mitotic activity >5/50 high-powered fields (HPF) were present in 6 of 73 (8%) and 5 of 73 (7%), respectively. Epithelioid areas were present in 33 of 73 (45%) and rosettes in 22 of 73 (30%). Follow up (54 cases; range, 2-192 mos; median, 24 mos; mean, 38 mos) showed 5 recurrences, 3 metastases, and 1 death. The diagnosis of LGFMS or HSCT was made prospectively in 51 patients; none had metastatic disease. Two of the metastatic tumors were LGFMS and one was a HSCT. LGFMS may occur more often in the pediatric population and show a much wider histologic spectrum than previously thought. A significant number of LGFMS possess inconspicuous collagen rosettes characteristic of HSCT, indicating that these two tumors are ends of a common spectrum rather than distinct entities. HSCT, like LGFMS, are low-grade sarcomas with metastatic potential. The presence of focal areas of intermediate- to high-grade sarcoma does not portend a worse outcome in the short term. The better prognosis reflected in this study compared with previous ones might reflect the fact that all were initially diagnosed as sarcomas and treated with aggressive surgery. The fact that the only three patients to develop metastatic disease were patients whose LGFMS or HSCT was identified retrospectively supports this concept. PMID- 11023097 TI - Solid-pseudopapillary tumor of the pancreas: immunohistochemical localization of neuroendocrine markers and CD10. AB - To clarify the neuroendocrine differentiation and CD10 expression in solid pseudopapillary tumors (SPTs) of the pancreas, we performed immunohistochemical analysis in 19 such tumors, including one solid-pseudopapillary carcinoma (SPC), along with 20 pancreatic neuroendocrine tumors (PNTs), six acinar cell carcinomas (ACCs), and one pancreatoblastoma (PB). We used antisera directed against CD56, synaptophysin, protein gene product 9.5, the alpha-subunit of Go protein, chromogranin A, CD10, trypsin, chymotrypsin, various cytokeratins (CKs), CA19-9, vimentin, and alpha-1-antitrypsin (AAT). All SPTs exhibited immunoreactivity for CD56 and CD10, and 15 expressed other neuroendocrine markers focally with the exception of chromogranin A. Frequent clustering of synaptophysin-positive cells was noted. Two cases contained a peculiar nodule that cytomorphologically and immunohistochemically resembled PNT. CD10-positive cells were scarce in one SPC. PNTs were CD56-positive, but often with faint intensity, and staining for other neuroendocrine markers, including chromogranin A, was diffusely positive. CD10 was detected, mostly in a focal pattern, in five PNTs. Pan-CK, CK8, CK18, and CK19 were more frequently demonstrated in PNT than SPT. Vimentin and AAT were often identified in PNT as well and were not specific for SPT. ACCs were CD56 negative, with the exception of one case designated as a mixed acinar-endocrine carcinoma. PB was focally positive for CD56 at the periphery of the tumor nests. Four ACCs and one PB exhibited focal CD10 reactivity. This study demonstrated the unique immunohistochemical features of SPT. Our results also suggest that SPT exhibits, at least focally, neuroendocrine differentiation, and that these neuroendocrine markers and CD10 are diagnostically useful. PMID- 11023098 TI - Intraductal papillary mucinous tumors of the pancreas confined to secondary ducts show less aggressive pathologic features as compared with those involving the main pancreatic duct. AB - Intraductal papillary mucinous tumors (IPMTs) of the pancreas are rare tumors characterized by a malignant potential. Because of the progress of imaging procedures, smaller cystic pancreatic lesions are now detected and some of them correspond to IPMTs that involve ectatic pancreatic branch ducts but spare the main pancreatic duct. To investigate differences in morphology and clinical behavior of branch and main duct types of IPMT, a surgical series of 43 cases was studied. All pathologic specimens of IPMT, surgically resected in our institution between October 1987 and July 1998, were analyzed. In all cases, the entire pancreatic specimen was systematically examined. IPMT of the branch type was found in 13 (30%) patients, whereas IPMT of main pancreatic duct type that involved the main pancreatic duct and branch ducts was observed in 30 (70%) patients. Patients with IPMT of the branch type were younger (median age, 55 yrs vs 64 yrs), and all but one of the lesions were located in the head and neck of the pancreas (vs 17 of 30 patients with the main duct type). The size of the cysts ranged from 4 to 55 mm, and the major duct showed a mild dilation in most cases. In contrast to the main pancreatic duct type, which showed invasive carcinoma and in situ carcinoma in 11 (37%) of 30 patients and 6 (20%) of 30 patients, respectively, IPMT of the branch type showed significantly less aggressive histologic lesions with five (39%) patients with simple hyperplasia, six (46%) patients with atypical hyperplasia, and two (15%) patients with in situ carcinoma. No invasive carcinoma was observed in this group. IPMT of the branch type occurs in younger patients and is associated with less aggressive histologic features than is the main pancreatic duct type. Our findings raise the difficult issue of clinical management of IPMT of the branch type as a distinctive group. PMID- 11023099 TI - Infarct of the prostate gland: experience on needle biopsy specimens. AB - Prostatic infarcts are uncommon and in the past have only been reported on transurethral resections of the prostate. We reviewed 13 consults and 2 nonconsult cases of needle biopsies showing prostatic infarcts from two institutions. The incidence of infarcts on biopsy were 2 in 2958 (0.07%) and 1 in 108,586 (0.0009%) in our nonconsult cases. Men averaged 71 years of age (range, 57-84 yrs). No relationship was seen with histories of hypertension, diabetes, atherosclerotic coronary vascular disease, recent surgery, and steroid use. Four of 12 men with available information had acute urinary retention, with markedly enlarged prostates in three (90 cc, 92 cc, 94 cc); two of these men had hematuria. An additional two men also had large glands (84 cc, 150 cc), one also with hematuria. Of eight men without acute urinary retention, three had sudden prostate-specific antigen (PSA) rises (increases of 199 ng/mL, 219 ng/mL, 287 ng/mL). Infarcts were usually an isolated focus on one core and varied from 1 mm to 11 mm (mean, 6.3 mm). Six cases showed earlier-aged infarcts with coagulative necrosis and recent hemorrhage and six showed intermediate-aged infarcts with reactive stroma and epithelium without necrosis. In the remaining three cases, there were remote infarcts characterized by replacement of the stroma by dense fibrosis with metaplastic glands. Adjacent tissue revealed reactive nests of immature squamous metaplasia in 14 of 15 cases with visible nucleoli (12 cases), squamous atypia (7 cases), and mitoses ranging from 1-10 (7 cases). Pathologists sent in 10 of 13 consult cases (77%) for problems with interpretation of the infarcts; remaining consults had other pathology of concern. One case was misdiagnosed as urothelial cancer. Features helpful in recognizing infarcts' benign nature were cyst formation containing cellular debris with or without neutrophils (73%), corpora amylacea (20%), and rings of collagen around squamous islands (40%). Infarcts are typically, although not exclusively, found in large prostates and may result in sudden rises in serum PSA. Infarcts' distinctive histology must be recognized and distinguished from necrosis resulting from infection and prior cryotherapy, as we have seen such misdiagnoses. Pathologists' awareness of prostatic infarcts on needle biopsy and their potential for atypical histology can prevent the misdiagnosis of cancer. PMID- 11023100 TI - Unicystic ameloblastoma: a clinicopathologic study of 33 Chinese patients. AB - The term unicystic ameloblastoma refers to those cystic lesions that show clinical, radiographic, or gross features of a jaw cyst, but on histologic examination show a typical ameloblastomatous epithelium lining part of the cyst cavity, with or without luminal and/or mural tumor growth. To ascertain the clinicomorphologic spectrum and biologic behavior of this tumor group, the clinicopathologic features of 33 unicystic ameloblastomas from Chinese patients were studied. This series represents approximately 19% of all cases of ameloblastoma accessioned in the authors' hospital during a 15-year period. Twenty-one patients were male and 12 were female, for a total of 33 patients. The age at diagnosis ranged from 8 to 60 years (mean, 25.3 yrs) and peaked at the second and third decades (70%), Thirty tumors (91%) occurred in the mandible and three in the maxilla. Of the 29 patients with a radiographic record, an expansive unilocular radiolucency was seen in 22 cases, and was multilocular in seven cases. Microscopically, all tumors demonstrated a generally monocystic growth pattern. Eight tumors were simple cystic, 10 comprised intraluminal tumor nodules, and the remaining 15 had a conspicuous component of infiltrative tumor islands in the cyst capsule. The cystic tumor linings invariably showed, at least in part, a typical ameloblastomatous pattern that was often accompanied by epithelial areas of various histologic appearance. Follow up of 29 patients revealed no recurrence in less than 4 years of follow up, but did reveal a 35% recurrence rate at more than 4 years of follow up. The average interval to recurrence was approximately 7 years. Recurrence also appeared to relate to histologic subtypes of unicystic ameloblastoma, with those invading the fibrous wall having a rate of 35.7%, but other types having a rate of 6.7%. Despite the fact that unicystic ameloblastoma may, in general, compare favorably with its solid or multicystic counterpart in terms of clinical behavior and response to treatment, the subsets of the maxillary lesions or tumors containing invading islands in the fibrous wall could have a high risk of recurrence. Furthermore, recurrence of unicystic ameloblastoma may be long delayed, and a long-term postoperative follow up is essential to the proper management of these patients. PMID- 11023101 TI - MIB-1 immunostaining is a beneficial adjunct test for accurate diagnosis of vulvar condyloma acuminatum. AB - The histopathologic diagnosis of vulvar condyloma acuminatum is often based on architectural features that are not specific for human papillomavirus (HPV) infection. Because HPV-associated lesions show increased cellular proliferation, the authors evaluated the usefulness of MIB-1 immunostaining as an aid in the differential diagnosis of cases equivocal for condyloma. The MIB-1 immunostaining pattern was correlated with HPV DNA detection in condyloma acuminatum (CON-A; n = 15), "consistent with condyloma" (c/w CON-A; n = 26), fibroepithelial polyp (FEP; n = 14), and squamous papilloma (n = 10). HPV was detected in 100% of the CON-A cases, and all cases demonstrated MIB-1-positive nuclei in the upper two thirds of the epithelial thickness. With this definition of MIB-1 positivity, there was complete concordance between MIB-1 positivity and HPV detection for all cases (kappa = 0.88). Of the cases c/w CON-A, 17 of 26 (65%) were positive for both MIB 1 and HPV, and could be reclassified as CON-A, whereas 35% were identified as an overdiagnosis based on negative results. In addition, two cases of FEP were MIB-1 and HPV positive, and thus were identified as an underdiagnosis. These results suggest significant overdiagnosis of cases equivocal for condyloma, and indicate that MIB-1 immunostaining is a beneficial adjunctive test when the morphologic features are suggestive but not diagnostic for CON-A. PMID- 11023102 TI - The histologic spectrum of epidermodysplasia verruciformis. AB - The classic histologic presentation of epidermodysplasia verruciformis is a verruca plana-type lesion with minimal hyperkeratosis and acanthotic areas where the cells contain perinuclear halos and blue-gray pallor. Whereas these lesions have a high malignant potential, it is important to elucidate the histologic spectrum of this entity and to differentiate it from its mimics. Fifteen skin biopsies from people with multiple cutaneous warts clinically suspicious for epidermodysplasia verruciformis were analyzed both histologically and for human papillomavirus (HPV) deoxyribonucleic acid (DNA) by in situ hybridization. Ten of the lesions contained HPV DNA, either type 5 (n = 6), type 8 (n = 3), or type 51 (n = 1). Only three of these lesions showed typical verruca plana. The histologic marker of HPV DNA in the other seven viral-positive cases was rare perinuclear halos in association with an irregular granular layer. The other five cases, which were also negative for viral DNA after polymerase chain reaction in situ hybridization, rarely demonstrated the abrupt variation in keratohyaline granules and concomitant perinuclear halos. The authors conclude that there is a wide spectrum of histologic changes in epidermodysplasia verruciformis and that viral testing in conjunction with the histologic and clinical findings can differentiate this premalignant entity from its mimics. PMID- 11023103 TI - Diffuse duodenitis associated with ulcerative colitis. AB - Backwash ileitis and postcolectomy pouchitis are well-recognized complications of ulcerative colitis (UC), whereas inflammation of the proximal small intestine is not. In contrast, small intestinal disease at any level is common in Crohn's disease (CD). Despite this well-established and accepted dogma, rare cases of histologically proven diffuse duodenitis (DD) associated with UC appear in the literature. In this study, we report our experience with similar cases exhibiting this unusual inflammatory phenomenon. Routine histologic sections from four cases of DD associated with well-documented UC were reviewed and the findings correlated with all available medical records. Multiple endoscopic biopsies showing histologic features of UC and colectomy specimens confirming severe ulcerative pancolitis were available for all cases. Varying degrees of active chronic inflammation and architectural mucosal distortion identical to UC were observed in pre- and postcolectomy duodenal biopsies of one of four and four of four cases, respectively. Similar inflammatory patterns were present postoperatively in the ileum in three of four cases and in the jejunum in one case. Endorectal pull-through (ERPT) procedures were performed in three of four patients and an end-to-end ileorectal anastomosis was done in one patient. Despite extensive upper gastrointestinal tract involvement, none of the patients developed postsurgical Crohn's-like complications during a follow-up period of 12 to 54 months. This suggests that patients with pancolitis and DD do not necessarily have CD, but rather may have UC and, most importantly, that successful ERPT procedures may be performed in these patients. PMID- 11023104 TI - Stratified mucin-producing intraepithelial lesions of the cervix: adenosquamous or columnar cell neoplasia? AB - BACKGROUND: Squamous (CIN) and glandular (ACIS) intraepithelial lesions often coexist in the same cervical specimen. However, a less common and little studied variant consists of a stratified epithelium resembling CIN in which conspicuous mucin production is present (Stratified Mucin-producing Intraepithelial LEsions (SMILE). This report describes the phenotypic characteristics of the SMILE, its associated lesions, and its immunophenotype. METHODS: Eighteen SMILEs were identified by the presence of conspicuous cytoplasmic clearing or vacuoles in lesions otherwise resembling CIN. The morphologic spectrum of SMILEs was detailed; including associated intraepithelial and invasive cervical neoplasms. In addition, selected cases were stained for mucicarmine, markers of squamous cell/reserve cell differentiation (keratin-14 and p63), and proliferative activity (Mib-1). RESULTS: Stratified neoplastic epithelial cells with a high Mib 1 index and a rounded or lobular contour at the epithelialstromal interface characterized SMILEs. In contrast to CIN, in which mucin droplets are confined to surface cells, mucin was present throughout the epithelium, varying from indistinct cytoplasmic clearing to discrete vacuoles. SMILEs were distinguished from benign metaplasia by nuclear hyperchromasia and a high Mib-1 index. All but three coexisted with either a squamous (CIN) or glandular (ACIS) precursor lesion. Nine of nine coexisting invasive carcinomas contained glandular, adenosquamous differentiation, or both. SMILEs stained negative for keratin-14 and variably for p63. When present, staining with p63 was confined to basal areas of SMILEs and was absent in areas of columnar differentiation. CONCLUSIONS: SMILEs are unusual cervical intraepithelial lesions best classified as variants of endocervical columnar cell neoplasia based on immunophenotype. The distribution and immunophenotype of SMILEs are consistent with a neoplasm arising in reserve cells in the transformation zone. The coexistence of a wide spectrum of intraepithelial and invasive cell phenotypes suggests that SMILEs are a marker for phenotypic instability, emphasizing the importance of identifying SMILEs and ensuring a complete examination of specimens containing this unusual precursor lesion. PMID- 11023105 TI - Stromal tumor of the gallbladder with phenotype of interstitial cells of Cajal: a previously unrecognized neoplasm. AB - We report a small, well-demarcated stromal tumor of the gallbladder in a 69-year old woman. The tumor and associated cholelithiasis led to chronic cholecystitis symptoms. The wall of the gallbladder contained a 2.4-cm hypocellular nodule composed of bland spindle-shaped cells that were immunoreactive for vimentin, CD34, and CD117. With the latter antibody, which stains interstitial cells of Cajal (ICC), the neoplastic cells appear fusiform with elongated bipolar projections or dendritic-like cytoplasmic projections. The gallbladder wall adjacent to the tumor contained numerous CD117-positive cells in close contact with the normal smooth muscle cells, whereas two of 10 gallbladders with minimal chronic cholecystitis showed only a few CD117-positive cells. These findings provide evidence that this stromal tumor of the gallbladder shows ICC differentiation similar to some stromal tumors of the gut. The presence of numerous ICC in the uninvolved gallbladder wall suggests that this tumor might have evolved through hyperplasia of ICC. PMID- 11023106 TI - Large cell neuroendocrine carcinoma of the gallbladder: report of two cases. AB - We report two cases of primary large cell neuroendocrine carcinoma (LCNEC) of the gallbladder, which, to the best of our knowledge, represent the first description of this entity. One of the tumors consisted entirely of LCNEC, whereas the second tumor was composed of LCNEC and the more common intestinal-type adenocarcinoma. Both tumors were morphologically similar to their pulmonary counterpart and were characterized by large cells with prominent nucleoli, coarse chromatin, and a high mitotic rate. The cells showed an organoid growth pattern with rosette formation and frequent areas of necrosis. Panendocrine markers were expressed in a variable proportion of tumor cells in both cases, and one of the cases also showed focal positivity for type 2 somatostatin receptors. One of the tumors followed a rapidly fatal course despite aggressive surgical treatment and chemotherapy administration, and the second patient is still alive and disease free 12 months after surgery. The description of these two cases of LCNEC of the gallbladder is significant for two reasons. From an academic standpoint, we now know that all the neuroendocrine tumors described in other organs can arise de novo in the gallbladder. More importantly, however, the recognition of this rare tumor type carries important clinical implications in regard to the use of chemotherapeutic agents and supplemental treatments (for example, somatostatin analogs). PMID- 11023107 TI - Pigmented liver cell adenoma in two male patients. AB - We report two cases of hepatocyte neoplasia with extensive deposition of Dubin Johnson-like pigment in men without Dubin-Johnson syndrome. This pigment has previously been described in hepatocellular carcinoma but not in liver cell adenoma. The tumors of both patients showed some atypical cytologic features, but no frank histologic evidence of malignancy. Long-term follow up for several years showed no evidence of recurrence after limited surgical excision. We conclude that tumors with this structure may be cured by limited surgical excision and should be considered as pigmented liver cell adenomas. PMID- 11023108 TI - Tubulocystic clear cell adenocarcinoma arising within the prostate. AB - Neoplasms resembling ovarian common epithelial-type tumors, including clear cell adenocarcinomas, rarely occur in the lower urinary tract of men. When they do, they develop in the urethra or urinary bladder. We report a case of such a tumor arising within the prostate of a 47-year-old man. The tumor was a cystic mass in the left posterolateral region of the prostate. Histologically, the tumor was chiefly composed of tubulocystic and papillary glands lined by glycogen-rich, cuboidal or hobnail cells with clear to eosinophilic cytoplasm. The tumor cells were strongly positive for pan-cytokeratin, low molecular weight cytokeratin, and epithelial membrane antigen, and focally positive for high molecular weight keratin. The tumor did not immunohistochemically express prostate-specific antigen (PSA) and prostatic acid phosphatase. Serologically, the patient had increased levels of CA125 instead of PSA. The clinical as well as the pathologic features are consistent with a clear cell adenocarcinoma as seen in the female genital tract rather than a typical prostatic adenocarcinoma. PMID- 11023109 TI - Chronic myeloproliferative disorders with myeloid metaplasia. PMID- 11023110 TI - Histopathologic features of malignant peripheral nerve sheath tumor are not restricted to metastatic malignant melanoma and can be found in primary malignant melanoma also. PMID- 11023111 TI - Dedifferentiation in salivary gland carcinomas. PMID- 11023112 TI - Deciduoid epithelial mesothelioma of the pleura with focal rhabdoid change. PMID- 11023113 TI - Gleason grading of prostatic biopsies. PMID- 11023114 TI - Pathologists should retain their individuality! PMID- 11023115 TI - Bone ageing: genetics versus environment. AB - Bone ageing results from a complex interaction between genetic and environmental factors (such as diet, climate and physical exercise) throughout human life. According to current literature, the most popular measures of bone ageing are osseometric measurements (OSM), bone mineral density (BMD) and osseographic scores (OSS), based on descriptive criteria of bone age. Plain roentgenography allows simultaneous assessment of all three measures. Ethnic differences with regard to these bone ageing characteristics have prompted us to study to the process anew, with the aim of elucidation the nature of the genetic and environmental components involved, and the possible interaction(s) between them. Despite abundant data on ethnic differences regarding these measures, modern knowledge on the genetics of these processes has derived primarily from the family studies of BMD, which pointed to strong involvement of the familial factors on bone mass. Segregation analysis performed by us in two ethnically different samples of pedigrees revealed a significant effect of the putative major gene on BMD of both compact and cancellous bone. The major finding of our bivariate segregation analysis was that it lead to the acceptance of the hypothesis predicating a single major locus with pleiotropy to both cancellous and compact BMD, but clearly rejecting the polygenic hypotheses. Our study of cortical index (CI) provided evidence that a single potential major gene controls not only the baseline trait level, but also the age at onset of the involutive bone changes, and the rate of the CI change with age. When we examined the environmental vs genetic influences on OSS variation in 32 human populations, we found very little environmental effect on the rate of bone change (r2 = 0.107), but a substantial effect on this rate of the genetic differences between populations (r = 0.480). Clarification of the genetic basis of bone ageing could have wide-ranging applications in the prevention and treatment of bone degenerative diseases such as osteoporosis and osteoarthritis, before irreversible damage takes place. There is thus a need to target the genetic analysis of BMD and the biochemical regulating factors of bone turnover through the use of molecular genetic techniques. PMID- 11023116 TI - The origins of the Irish travellers and the genetic structure of Ireland. AB - Ireland's unique and well-documented history provides insight into the formation and origins of population subdivisions. Of particular interest, is the controversial ethnogenesis of an itinerant population of Ireland: the Travellers. The objectives of this study were: (1) to determine the genetic affinity of the Travellers to the general Irish population based on gene frequency data, subdivided by county, and (2) to explore the relationship between subpopulations of Ireland, given its turbulent history. The gene frequencies of standard genetic markers collected from populations residing in counties of Ireland and the Travellers were calculated and analysed using several multivariate methods. First, a relationship (R) matrix was used to ascertain the scaled variance covariance matrix of population similarity. Second, mean per locus heterozygosity (H) was regressed on distance of the region from the gene frequency centroid (r(ii)). The results of this study include: (1) the confirmation of Crawford's (1975, in Biosocial Interrelations in Population Adaptations, E. S. Watts et al. (eds), pp. 93-103) conclusions concerning the origins and genetic affinity of the Travellers; (2) based on several multivariate analyses, the major influence on population structure was unique historical events; and (3) Relethford and Crawford's (1995, American Journal of Physical Anthropology, 96, 25-38) hypothesis concerning the distinctiveness of the midland counties was verified by this study. PMID- 11023117 TI - Familial resemblance for physique: heritabilities for somatotype components. AB - PRIMARY OBJECTIVE: To examine familial resemblance in the Heath-Carter anthropometric somatotype in a sample of 328 participants from 103 nuclear families in Northern Ontario (Canada). METHODS AND PROCEDURES: The three somatotype components (endomorphy, mesomorphy, ectomorphy) were subjected to principal components analysis and the resulting first principal component (PCI) was used as an additional index of physique. The four phenotypes were adjusted for age, sex and generation effects, while each of the three somatotype components was further adjusted for the effects of the other two components using regression procedures. A familial correlation model was fit to the data and used to estimate the degree of familial resemblance in somatotype. MAIN OUTCOME AND RESULTS: For all somatotype variables, the most parsimonious model was one in which there was no spouse resemblance and no sex or generation effects in the familial correlations. Maximal heritabilities were 56%, 68%, 56% and 64% for endomorphy, mesomorphy, ectomorphy and PCI, respectively, indicating significant familial resemblance for the Heath-Carter anthropometric somatotype. Further, the pattern of familial correlations suggests the role of genetic factors in explaining variation in human physique. CONCLUSIONS: In general, a pattern of no spouse but significant parent-child correlations implicates the role of genes on human physique, provided that mating is random with regard to these traits. PMID- 11023118 TI - The effect of type of physical activity measure on the relationship between body fatness and habitual physical activity in children: a meta-analysis. AB - BACKGROUND: The relationship between activity levels and body fat in children is unclear, despite a large number of studies. The issue is clouded by the wide variety of methods used to assess children's activity levels. It is important to assess whether the type of activity measure influences the fatness-activity relationship. This is a first step to uncovering the role of modifying variables such as gender, age, maturity, etc. PRIMARY OBJECTIVE: This study uses meta analytic procedures to synthesize the results of such studies and to assess whether the type of activity measure used has an effect on the strength of the relationship observed. METHODS AND PROCEDURES: Fifty studies were located that satisfied the inclusion criteria. Seventy-eight per cent of the studies showed a negative relationship, 18% no relationship and 4% a positive relationship between physical activity and body fatness. Data were analysed using the meta-analytic procedures described by Rosenthal (Meta-analytic Procedures for Social Research, Sage, 1991). MAIN OUTCOMES AND RESULTS: The mean effect size indicated a small to moderate, inverse relationship (r = -0.16). Mean effect sizes differed significantly (F(3,52) = 8.04, p < 0.001) according to the activity measure used: questionnaire, r = -0.14; motion counters, r = -0.18; observation, r = -0.39; heart rate (HR), r = 0.00. Observation measures elicited a significantly stronger relationship with body fat than did questionnaire or heart rate measures (p < 0.05). However, there was no significant difference between the effect sizes elicited by observation and motion counters. Correlational analyses revealed no effect of age group or gender on the strength of the relationship between fatness and activity. CONCLUSIONS: This meta-analysis suggests there is a small to moderate relationship between body fat and activity in children. It is important to note, however, that the size of the relationship depends on the activity measure used. It is therefore recommended that direct measures of movement, such as observation or motion counter methods, are used to assess the relationship of activity levels with health. PMID- 11023119 TI - Is fasting insulin associated with blood pressure in obese children? AB - PRIMARY OBJECTIVE: We tested whether fasting insulin levels are associated with blood pressure in a large sample of obese children. SUBJECTS AND METHODS: Three hundred and fifty obese children (F:M ratio = 1.03) of 10.1 +/- 2.7 y of age (mean +/- SD) were consecutively enrolled at an Outpatient Paediatric Clinic. Obesity was diagnosed on the basis of a relative weight for age > 120% and hypertension on the basis of a systolic (SBP) or diastolic (DBP) blood pressure > 95th percentile for age after adjustment for height (Ht). MAIN OUTCOME AND RESULTS: Insulin was significantly higher in hypertensive (n = 202, 58%) than normotensive (n = 148, 42%) children (16 vs 14 microU mL(-1), geometric mean, p < 0.01, ANOVA) but the difference was not clinically relevant. Moreover, (log transformed) insulin explained only 7 and 4% of SBP and DBP variance, respectively (p < 0.0001 for both) and this contribution disappeared after the confounding effects of age, weight or other anthropometric dimensions were taken into account (p = ns, ANCOVA). CONCLUSIONS: This study does not support the hypothesis of a clinically relevant association between fasting insulin and blood pressure in obese children. PMID- 11023120 TI - Anthropometry of adolescent girls in Bahrain, including body fat distribution. AB - PRIMARY OBJECTIVE: To determine anthropometric measurements of adolescent Bahraini girls, including obesity and fat composition. METHODS AND PROCEDURES: A cross-sectional sample of 584 Bahraini girls aged 12-19 years, were selected from schools using multistage stratified sampling procedure. Fifteen anthropometric measurements were taken (weight, height, circumferences for upper arm, upper forearm, upper chest, chest, waist, hip, thigh and medial calf, triceps, biceps, subscapular and suprailiac). Body mass index (BMI), waist/hip ratio, sum of skinfold thickness and % body fat were also computed. MAIN OUTCOMES AND RESULTS: The findings revealed a significant increase in all anthropometric measurements with increase in age. The mean weights for girls was higher than those reported in 1986 for the same age group, but no difference was observed in the mean heights, indicating a trend toward overweight. Using the 85th and 95th centiles of the National Health and Nutrition Examination Survey I (NHANESI) BMI distribution to define respectively overweight and obesity, the prevalence of overweight was 38.5% and of obesity was 6.3%. The means for BMI, waist/hip ratio, sum of skinfold thickness and % body fat were higher than those reported in many developed and developing countries. CONCLUSIONS: Bahraini adolescent girls have a higher proportion of body fat than their counterparts in many Western countries. This may contribute to some chronic diseases in adulthood. An intervention programme, therefore, is urgently needed to reduce overweight and obesity at childhood and adolescent stages. PMID- 11023121 TI - Non-seasonality of births in Tierra del Fuego (Chile). AB - Tierra del Fuego is situated at the southern tip of the American continent, which conditions its environmental and climatic characteristics. The colonizing population arrived, at the end of the 19th century, from other Chilean regions (particularly from Chiloe) and diverse European countries, especially Britain and Croatia, but also Germany, Spain and Italy. In the present study, the existence of a seasonal pattern in 5430 births registered in the Chilean population of Tierra del Fuego from 1890 to 1995 was analysed. The analysis showed no seasonal distribution of births in the periods 1890-1920 and from 1946 to the present day, a phenomenon rarely reported in the literature. The absence of seasonality in birth distribution could be related to the great diversity in the origins of the population's families, the constant renovation of this and the declining importance of the administrative capital of the province of Tierra del Fuego, Porvenir, in favour of Punta Arenas, capital of the Magellanic region. In the period of maximum development of the capital of the province, a seasonal pattern was detected with a peak in April and a trough in October, which corresponds with a maximum of conceptions in July and, in general, during the early southern winter and a decline in births from late spring to mid-summer, with a trough in January. This birth distribution is interpreted as a subordination of activity and social life to the annual sheep cycle. In addition to temporal trends, the influence on the observed patterns of environmental parameters, father's occupation, seasonality of marriage and the origin of the parents were analysed. PMID- 11023122 TI - What is your diagnosis? Thoracic trauma. PMID- 11023123 TI - Polyneuropathies of cats. AB - Polyneuropathies of cats have a variety of clinical presentations. Areflexic flaccid quadriparesis, or quadriplegia, progressing over a 24- to 48-hour period, may be associated with polyneuropathies, as can chronic insidiously progressive tremors and muscle weakness that wax and wane or progress slowly over weeks or months, and which can go undiagnosed for years. In addition, these neurological signs may be due to spinal cord, neuromuscular junction or muscle disorders, so the diagnosis of polyneuropathy can be a challenge even for the most astute of clinicians. Polyneuropathies may have congenital, inherited, inflammatory, metabolic and toxic causes. Sometimes the underlying aetiology is not found and a diagnosis of idiopathic polyneuropathy is made. Since the treatment and prognosis of polyneuropathies in cats vary, the purpose of this review is to assist the veterinary practitioner to recognise, appropriately manage and provide an accurate prognosis for these challenging cases. PMID- 11023124 TI - Preliminary clinical observations on the use of piroxicam in the management of rectal tubulopapillary polyps. AB - Rectal tubulopapillary polyps were diagnosed in eight dogs following proctoscopy and mucosal pinch biopsy. Histological examination of the pinch biopsies revealed evidence of malignant transformation in three of the cases. The remaining cases were diagnosed as benign polyps. Inflammatory changes were observed in four cases. Seven dogs were treated with piroxicam suppositories and one with oral piroxicam. All dogs were re-examined after four to six weeks of piroxicam therapy and the extent of haematochezia, tenesmus and faecal mucus production was reduced in all cases. The owners of seven of the dogs considered the improvement in clinical signs to be good or excellent. Cases with and without evidence of inflammation responded equally well. This finding supports the hypothesis that piroxicam has an antineoplastic effect due to apoptosis and alteration in the cell cycle. Medical management with piroxicam may provide a non-invasive treatment option for dogs with rectal polyp formation in which surgical treatment is likely to be associated with complications such as incontinence, infection and wound breakdown, or where the owner declines such treatment. PMID- 11023125 TI - Middle ear disease associated with congenital palatine defects in seven dogs and one cat. AB - Medical records of eight dogs and one cat with congenital palatine defects were reviewed retrospectively. Five of the dogs had nasal discharge and seven had radiographic signs of middle ear disease, but no clinical signs of ear disease were identified in any of the dogs, nor were any reported by their owners during a one- to five-year follow-up period. One dog had an ipsilateral impairment of hearing detected by brainstem auditory evoked responses. The cat had clinical and radiographic signs of middle ear disease. These findings suggest that, as in humans, congenital palatine defects in dogs and cats may predispose to middle ear disease. Any associated deafness could cause problems for working dogs. PMID- 11023126 TI - Glucagon-producing neuroendocrine tumour associated with hypoaminoacidaemia and skin lesions. AB - A nine-year-old neutered female crossbred Bernese mountain dog was diagnosed with superficial necrolytic dermatitis and a glucagon-producing islet cell tumour. Laboratory findings included hyperglucagonaemia and hypoaminoacidaemia. The dog was euthanased because of progression of the disease, and necropsy revealed liver metastases of a neuroendocrine carcinoma with immunohistochemical expression of glucagon and somatostatin. This report represents a case of canine glucagonoma syndrome; the previously reported cases in dogs are also briefly described. PMID- 11023127 TI - Persistent cobalamin deficiency causing failure to thrive in a juvenile beagle. AB - A six-month-old beagle was presented with a three-month history of failure to gain weight, lethargy, intermittent vomiting and seizures. Hypoglycaemia, portosystemic shunt, lead intoxication, gastrointestinal diseases and hereditary metabolic disorders were considered. Laboratory test results of low serum cobalamin (Cbl) concentrations, anaemia, leucopenia and methylmalonic aciduria while the dog was receiving a balanced commercial canine diet were suggestive of a congenital selective Cbl malabsorption. Treatment with repeated injections of parenteral cyanocobalamin (CN-Cbl) at 50 microg/kg every two weeks corrected the Cbl-deficient state and reversed all the clinical abnormalities. Selective Cbl malabsorption has previously been described in giant schnauzers and border collies and represents a unique readily treatable hereditary metabolic disorder. PMID- 11023128 TI - Intestinal haemangiosarcoma in the cat: clinical and pathological features of four cases. AB - The clinical and pathological features of four cases of feline intestinal haemangiosarcoma are described. All cases were in domestic shorthaired cats and the mean age of the animals (n=3) was 13 years. The tumours originated in the colon, small intestine, ileocaecocolic junction or rectum. The rectal tumour was juxtaposed with an adjacent mast cell neoplasm. Metastasis to mesenteric lymph node occurred in two cases, and in one of these cats there was also abdominal seeding. The histopathological appearance was of a spindle cell neoplasm with vascular differentiation in each case. Immunohistochemical staining for factor VIII-related antigen, an endothelial cell marker, confirmed all four tumours to be of endothelial origin. The neoplastic endothelial cells lining irregular vascular channels were more likely to express the antigen than those forming denser sheets without obvious vascular differentiation. PMID- 11023129 TI - Idiopathic phenobarbital-responsive hypersialosis in the dog: an unusual form of limbic epilepsy? AB - Three unusual cases of salivary gland enlargement and hypersialosis in the dog that responded to anticonvulsant therapy are reported. Presenting complaints included weight loss, hypersalivation, retching and vomiting of several weeks' duration. Two dogs were presented with enlarged painful mandibular salivary glands. The third dog exhibited bizarre behaviour (including jaw chattering) and developed enlarged painful mandibular salivary glands during hospitalisation. Fine needle aspirate cytology and biopsies from the enlarged salivary glands revealed no significant pathological changes. In one dog, an electroencephalogram revealed changes consistent with epilepsy. Hypersialism and salivary gland enlargement resolved completely during phenobarbital administration in all cases. Two dogs were successfully weaned off treatment six months after diagnosis. The remaining dog relapsed after eight months, but normalised with the addition of oral potassium bromide. It is hypothesised that the syndrome idiopathic hypersialosis may in fact be an unusual form of limbic epilepsy. PMID- 11023130 TI - Renal dysplasia in boxers and Finnish harriers. AB - Puppies from two litters of dogs were found to have severe polyuria and polydipsia. Four of the dogs were investigated by means of clinical examination, haematological and biochemical analysis, and urinalysis. A modified water deprivation response test was also performed in two of the dogs. Renal changes on postmortem examination in three of the dogs were found to be consistent with renal dysplasia. A possible explanation for the finding of hyposthenuria and the extreme polyuria and polydipsia in association with renal dysplasia may be lack of response to antidiuretic hormone owing to anomalous maturation of the renal tubules. Six other puppies from the two litters of dogs did not show any clinical signs of polyuria and polydipsia, although postmortem examination in one of them also revealed renal dysplasia. The clinical features of renal dysplasia may therefore vary greatly between individuals. PMID- 11023131 TI - Animal salivary glands. PMID- 11023132 TI - Blood transfusions. BSAVA's Scientific Committee. British Small Animal Veterinary Association. PMID- 11023133 TI - Strategies for urban animal management. PMID- 11023134 TI - Weight loss and mortality in type 2 diabetes. PMID- 11023135 TI - Giving smoking cessation the attention that it deserves. PMID- 11023136 TI - Intervention study for smoking cessation in diabetic patients: a randomized controlled trial in both clinical and primary care settings. AB - OBJECTIVE: To evaluate the effectiveness of a nurse-managed smoking cessation intervention in diabetic patients. RESEARCH DESIGN AND METHODS: This randomized controlled clinical trial involved 280 diabetic smokers (age range 17-84 years) who were randomized either into control (n = 133) or intervention (n = 147) groups at 12 primary care centers and 2 hospitals located in Navarre, Spain. The intervention consisted of a 40-min nurse visit that included counseling, education, and contracting information (a negotiated cessation date). The follow up consisted of telephone calls, letters, and visits. The control group received the usual care for diabetic smokers. Baseline and 6-month follow-up measurements included smoking status (self-reported cessation was verified by urine cotinine concentrations), mean number of cigarettes smoked per day, and stage of change. RESULTS: At the 6-month follow-up, the smoking cessation incidence was 17.0% in the intervention group compared with 2.3% in the usual care group, which was a 14.7% difference (95% CI 8.2-21.3%). Among participants who continued smoking, a significant reduction was evident in the average cigarette consumption at the 6 month follow-up. The mean number of cigarettes per day decreased from 20.0 at baseline to 15.5 at 6 months for the experimental group versus from 19.7 to 18.1 for the control group (P < 0.01). CONCLUSIONS: A structured intervention managed by a single nurse was shown to be effective in changing the smoking behavior of diabetic patients. PMID- 11023137 TI - Long-term dietary treatment with increased amounts of fiber-rich low-glycemic index natural foods improves blood glucose control and reduces the number of hypoglycemic events in type 1 diabetic patients. AB - OBJECTIVE: To evaluate in type 1 diabetic patients 1) the long-term feasibility of a high-fiber (HF) diet composed exclusively of natural foodstuffs and 2) the efficacy of this diet in relation to blood glucose control and incidence of hypoglycemic episodes. RESEARCH DESIGN AND METHODS: The study was randomized with parallel groups. Participants were part of a larger multicenter study on the effects of acarbose on glucose control in diabetes. A total of 63 type 1 diabetic patients, age 28 +/- 9 years, BMI 24 +/- 0.6 kg/m2, after a 4-week run-in period on their habitual diet, were randomized to either an HF (n = 32) or a low-fiber (LF) diet (n = 31) for 24 weeks. The two diets, composed exclusively of natural foodstuffs, were weight-maintaining and, aside from their fiber content, were similar for all nutrients. At the end of the run-in period and the dietary treatment, fasting blood samples for the measurement of plasma cholesterol, HDL cholesterol, triglyceride, and HbA(1c) were collected. A daily glycemic profile was performed on a day in which the participants had consumed a standard menu representative of their treatment diet (HF or LF). RESULTS: Of the 63 study subjects, 29 in the HF group (91%) and 25 in the LF group (81%) completed the study Compared with the LF diet, the HF diet after 24 weeks decreased both mean daily blood glucose concentrations (P < 0.05) and number of hypoglycemic events (P < 0.01). When compliance to diet was taken into account, 83% of the subjects on the HF diet and 88% on the LF diet were compliant. In this subgroup, compared with the LF diet, the HF diet significantly reduced mean daily blood glucose concentrations (P < 0.001), HbA(1c) (P < 0.05), and number of hypoglycemic events (P < 0.01). CONCLUSIONS: In type 1 diabetic patients, an HF diet is feasible in the long term and, compared with an LF diet, improves glycemic control and reduces the number of hypoglycemic events. PMID- 11023138 TI - Clinical characteristics of type 1 diabetic patients with and without severe hypoglycemia. AB - OBJECTIVE: To investigate the frequency of severe hypoglycemia (SH) and hypoglycemic coma and to identify clinical and behavioral risk indicators in a nonselected population of type 1 diabetic patients. RESEARCH DESIGN AND METHODS: This study involved a retrospective clinical survey of 195 consecutive patients using a questionnaire addressing the frequency of SH (i.e., help from others required) and hypoglycemic coma during the previous year, general characteristics, behavior, hypoglycemia awareness, and the Hypoglycemia Fear Survey Data regarding diabetes, treatment, long-term complications, comorbidity, and comedication were obtained from the patients' medical records. RESULTS: A total of 82% of subjects were receiving intensive insulin treatment, and mean +/- SD HbA(1c) was 7.8 +/- 1.2%. Mean duration of diabetes was 20 +/- 12 years. The occurrence of SH (including hypoglycemic coma) was 150 episodes/100 patient-years and affected 40.5% of the population. Hypoglycemic coma occurred in 19% of subjects (40 episodes/100 patient-years). SH without coma was independently related to nephropathy (odds ratio [OR] 4.8 [95% CI 1.5-15.1]), a threshold for hypoglycemic symptoms of <3 mmol/l (4.8 [1.8-12.0]), and a daily insulin dose 0.1 U/kg higher (1.3 [1.0-1.6]) (all ORs were adjusted for diabetes duration and use of comedication). Hypoglycemic coma was independently related to neuropathy (3.9 [1.5-10.4]), (nonselective) beta-blocking agents (14.9 [2.1-107.4]), and alcohol use (3.5 [1.3-9.1]) (all ORs were adjusted for diabetes duration). CONCLUSIONS: SH and hypoglycemic coma are common in a nonselected population with type 1 diabetes. The presence of long-term complications, a threshold for symptoms of <3 mmo/l, alcohol use, and (nonselective) beta-blockers were associated with SH during the previous year. If prospectively confirmed, these results may have consequences for clinical practice. PMID- 11023139 TI - Dietary unsaturated fatty acids in type 2 diabetes: higher levels of postprandial lipoprotein on a linoleic acid-rich sunflower oil diet compared with an oleic acid-rich olive oil diet. AB - OBJECTIVE: The present study was undertaken to examine the effect of a polyunsaturated fat diet compared with an isocaloric Mediterranean-style monounsaturated fat diet. RESEARCH DESIGN AND METHODS: This was a randomized 2 week crossover study on either a high-polyunsaturated or a high-monounsaturated fat diet in 11 well-controlled diabetic men. Blood was taken fasting and for up to 8 h after a high fat meal. Lipoproteins were isolated by sequential ultracentrifugation. Apolipoprotein (apo) B48 and apo B100 were separated by PAGE. Fatty acids were analyzed by gas-liquid chromatography RESULTS: Fasting blood glucose and insulin levels were significantly higher on the linoleic acid diet compared with the oleic acid diet (P < 0.01 and P < 0.002, respectively). Plasma cholesterol and LDL cholesterol levels were also significantly higher on the linoleic acid diet (P < 0.001). Likewise, fasting chylomicron apo B48 and apo B100 (P < 0.05) and postprandial chylomicron and VLDL apo B48 and B100 (P < 0.05) were also higher on the linoleic acid diet. CONCLUSIONS: This study suggests that, in type 2 diabetes, an oleic acid-rich Mediterranean-type diet versus a linoleic acid-enriched diet may reduce the risk of atherosclerosis by decreasing the number of chylomicron remnant particles. PMID- 11023140 TI - Effect of intensive glycemic control on microalbuminuria in type 2 diabetes. Veterans Affairs Cooperative Study on Glycemic Control and Complications in Type 2 Diabetes Feasibility Trial Investigators. AB - OBJECTIVE: Microalbuminuria can reflect the progress of microvascular complications and may be predictive of macrovascular disease in type 2 diabetes. The effect of intensive glycemic control on microalbuminuria in patients in the U.S. who have had type 2 diabetes for several years has not previously been evaluated. RESEARCH DESIGN AND METHODS: We randomly assigned 153 male patients to either intensive treatment (INT) (goal HbA(1c) 7.1%) or to standard treatment (ST) (goal HbA(1c) 9.1%; P = 0.001), and data were obtained during a 2-year period. Mean duration of known diabetes was 8 years, mean age of the patients was 60 years, and patients were well matched at baseline. We obtained 3-h urine samples for each patient at baseline and annually and defined microalbuminuria as an albumin:creatinine ratio of 0.03-0.30. All patients were treated with insulin and received instructions regarding diet and exercise. Hypertension and dyslipidemia were treated with similar goals in each group. RESULTS: A total of 38% of patients had microalbuminuria at entry and were evenly assigned to both treatment groups. INT retarded the progression of microalbuminuria during the 2 year period: the changes in albumin:creatinine ratio from baseline to 2 years of INT versus ST were 0.045 vs. 0.141, respectively (P = 0.046). Retardation of progressive urinary albumin excretion was most pronounced in those patients who entered the study with microalbuminuria and were randomized to INT. Patients entering with microalbuminuria had a deterioration in creatinine clearance at 2 years regardless of the intensity of glycemic control. In the group entering without microalbuminuria, the subgroup receiving ST had a lower percentage of patients with a macrovascular event (17%) than the subgroup receiving INT (36%) (P = 0.03). Use of ACE inhibitors or calcium-channel blockers was similarly distributed among the groups. CONCLUSIONS: Intensive glycemic control retards microalbuminuria in patients who have had type 2 diabetes for several years but may not lessen the progressive deterioration of glomerular function. Increases in macrovascular event rates in the subgroup entering without albuminuria who received INT remain unexplained but could reflect early worsening, as observed with microvascular disease in the Diabetes Control and Complications Trial. PMID- 11023141 TI - Circumscribed cognitive dysfunction in middle-aged adults with type 2 diabetes. AB - OBJECTIVE: To examine the extent to which type 2 diabetes is associated with poorer performance on measures of learning, memory, psychomotor speed, and problem-solving in middle-aged adults. RESEARCH DESIGN AND METHODS: This cross sectional study evaluated 50 adults (age range 34-65 years, mean 50.8) with type 2 diabetes and 50 demographically similar community control subjects without diabetes. Each subject received a thorough physical examination and a detailed neuropsychological assessment. Factor analysis was used to assign specific tests to 1 of 4 cognitive domains (learning, memory for stories, problem-solving, and psychomotor speed). Hierarchical regression analysis was used to identify demographic and biomedical variables associated with cognitive dysfunction. RESULTS: Learning, memory, and problem-solving skills were unaffected by type 2 diabetes. In contrast, psychomotor slowing was predicted by a diagnosis of diabetes (r2 change = 0.075, P < 0.002) with additional variance in psychomotor efficiency explained independently by HbA1 (r2 = 0.064, P < 0.003) and vibratory threshold (r2 = 0.112, P < 0.0001). The magnitude of psychomotor slowing on specific tests ranged from 12% (Digit Vigilance) to 23% (Grooved Pegboard). CONCLUSIONS: Middle-aged adults with type 2 diabetes manifest psychomotor slowing that is associated with poorer metabolic control, whereas learning, memory, and problem-solving skills appear to be largely intact. The development of psychomotor slowing may be a manifestation of a "central neuropathy" induced by chronic hyperglycemia. PMID- 11023142 TI - What degree of maternal metabolic control in women with type 1 diabetes is associated with normal body size and proportions in full-term infants? AB - OBJECTIVE: To assess what degree of maternal metabolic control in women with type 1 diabetes is associated with normal fetal growth and results in normal neonatal body proportions in a group of full-term infants. RESEARCH DESIGN AND METHODS: We investigated the anthropometric characteristics of 98 full-term singleton infants born to 98 Caucasian women with type 1 diabetes enrolled within 12 weeks of gestation. The type 1 diabetic mother-infant pairs were divided into three groups on the basis of the daily glucose levels reached during the second and third trimesters of pregnancy (group 1: 37 mother-infant pairs with an average daily glucose level during the second and third trimesters of < or =95 mg/dl; group 2: 37 mother-infant pairs with an average daily glucose level during the second trimester of >95 mg/dl and during the third trimester of < or =95 mg/dl; group 3: 24 mother-infant pairs with an average daily glucose level during the second and third trimesters of >95 mg/dl; control group: 1,415 Caucasian mother-infant pairs with full-term singleton pregnancies and normal glucose challenge test screened for gestational diabetes. RESULTS: Infants of diabetic mothers in group 1 were similar to those of the control group in birth weight and in other anthropometric parameters. In contrast, offspring of diabetic mothers of groups 2 and 3 showed an increased incidence of large-for-gestational-age infants, significantly greater means of ponderal index and thoracic circumferences, and significantly smaller cranial/thoracic circumference ratios with respect to the control group. CONCLUSIONS: The results of our study suggest that, in diabetic pregnancies, only overall daily glucose values < or =95 mg/dl throughout the second and third trimesters can avoid alterations in fetal growth. PMID- 11023143 TI - Intentional weight loss and mortality among overweight individuals with diabetes. AB - OBJECTIVE: To estimate the effect of intentional weight loss on mortality in overweight individuals with diabetes. RESEARCH DESIGN AND METHODS: We performed a prospective analysis with a 12-year mortality follow-up (1959-1972) of 4,970 overweight individuals with diabetes, 40-64 years of age, who were enrolled in the American Cancer Society's Cancer Prevention Study I. Rate ratios (RRs) were calculated, comparing overall death rates, and death from cardiovascular disease (CVD) or diabetes in individuals with and without reported intentional weight loss. RESULTS: Intentional weight loss was reported by 34% of the cohort. After adjustment for initial BMI, sociodemographic factors, health status, and physical activity, intentional weight loss was associated with a 25% reduction in total mortality (RR = 0.75; 95% CI 0.67-0.84), and a 28% reduction in CVD and diabetes mortality (RR = 0.72; 0.63-0.82). Intentional weight loss of 20-29 lb was associated with the largest reductions in mortality (approximately 33%). Weight loss >70 lb was associated with small increases in mortality CONCLUSIONS: Intentional weight loss was associated with substantial reductions in mortality in this observational study of overweight individuals with diabetes. PMID- 11023144 TI - Dental and other health care visits among U.S. adults with diabetes. AB - OBJECTIVE: This study compared yearly dental visits of diabetic adults with those of nondiabetic adults. For adults with diabetes, we compared the frequency of past-year dental visits with past-year visits for diabetes care, dilated eye examinations, and foot examinations. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional study using a sample of 105,718 dentate individuals aged > or =25 years, including 4,605 individuals with diabetes who participated in the 1995 1998 Behavioral Risk Factor Surveillance System in 38 states. RESULTS: Dentate adults (i.e., those with at least some natural teeth) with diabetes were less likely than those without diabetes to have seen a dentist within the preceding 12 months (65.8 vs. 73.1%, P = 0.0000). Adults with diabetes were less likely to have seen a dentist than to have seen a health care provider for diabetes care (86.3%); the percentage who saw a dentist was comparable with the percentage who had their feet examined (67.7%) or had a dilated eye examination (62.3%). The disparity in dental visits among racial or ethnic groups and among socioeconomic groups was greater than that for any other type of health care visit for subjects with diabetes. CONCLUSIONS: Promotion of oral health among diabetic patients may be necessary, particularly in Hispanic and African-American communities. Information on oral health complications should be included in clinical training programs. Oral and diabetes control programs in state health departments should collaborate to promote preventive dental services, and the oral examination should be listed as a component of continuous care in the American Diabetes Association's standards of medical care for diabetic patients. PMID- 11023145 TI - Effect of adults' self-regulation of diabetes on quality-of-life outcomes. AB - OBJECTIVE: To examine the relationships among cognitive representations of diabetes, diabetes-specific health behaviors, and quality of life using Leventhal and Diefenbach's self-regulation model of illness (Leventhal H, Diefenbach M: The active side of illness cognition. In Mental Representation in Health and Illness. SkeltonJA, Croyle RT, Eds. New York, Springer-Verlag, 1991, p. 247-272). RESEARCH DESIGN AND METHODS: This research involved secondary analysis of a mailed survey completed by 296 adults (ages 20-90 years). Structural equation modeling was conducted to investigate relationships among cognitive representations, diabetes specific health behaviors, and quality of life. Model differences by diabetes type were also investigated. RESULTS: Findings indicated that certain cognitive representation constructs were related to increased diabetes-specific health behaviors, decreased sense of burden, and positive quality-of-life outcomes. Individuals levels of understanding of diabetes and their perceptions of control over diabetes were the most significant predictors of outcomes. However, diabetes specific health behaviors were related to an increased sense of burden that was negatively associated with quality of life. Multigroup analyses indicated that this self-regulatory model provided a good fit for individuals with type 1 diabetes, those with type 2 diabetes who take insulin, and those with type 2 diabetes who do not take insulin. CONCLUSIONS: These findings advance what is known about cognitive representations of illness and the self-regulation of diabetes as well as the relationships between cognitive representations of illness, quality of life, and behavioral factors. In particular, results from this study suggest the need for further study to address ways of reducing the burden of diabetes associated with health behaviors and decreased quality of life. PMID- 11023146 TI - Incidence of childhood type 1 diabetes worldwide. Diabetes Mondiale (DiaMond) Project Group. AB - OBJECTIVE: To investigate and monitor the patterns in incidence of childhood type 1 diabetes worldwide. RESEARCH DESIGN AND METHODS: The incidence of type 1 diabetes (per 100,000 per year) from 1990 to 1994 was determined in children < or =14 years of age from 100 centers in 50 countries. A total of 19,164 cases were diagnosed in study populations totaling 75.1 million children. The annual incidence rates were calculated per 100,000 population. RESULTS: The overall age adjusted incidence of type 1 diabetes varied from 0.1/100,000 per year in China and Venezuela to 36.8/100,000 per year in Sardinia and 36.5/100,000 per year in Finland. This represents a >350-fold variation in the incidence among the 100 populations worldwide. The global pattern of variation in incidence was evaluated by arbitrarily grouping the populations with a very low (<1/100,000 per year), a low (1-4.99/100,000 per year), an intermediate (5-9.99/100,000 per year), a high (10-19.99/100,000 per year), and a very high (> or =20/100,000 per year) incidence. Of the European populations, 18 of 39 had an intermediate incidence, and the remainder had a high or very high incidence. A very high incidence (> or =20/ 100,000 per year) was found in Sardinia, Finland, Sweden, Norway Portugal, the U.K., Canada, and New Zealand. The lowest incidence (<1/100,000 per year) was found in the populations from China and South America. In most populations, the incidence increased with age and was the highest among children 10-14 years of age. CONCLUSIONS: The range of global variation in the incidence of childhood type 1 diabetes is even larger than previously described. The earlier reported polar-equatorial gradient in the incidence does not seem to be as strong as previously assumed, but the variation seems to follow ethnic and racial distribution in the world population. PMID- 11023147 TI - Diabetes is an independent risk factor for in-hospital mortality from acute spontaneous intracerebral hemorrhage. AB - OBJECTIVE: We tested the hypothesis that diabetes is an independent determinant of outcome after intracerebral hemorrhage (ICH). RESEARCH DESIGN AND METHODS: This was a hospital-based prospective study The setting was an acute care 350-bed hospital in the city of Barcelona, Spain. Spontaneous ICH was diagnosed in 229 (11%) of 2,000 consecutive stroke patients included in a prospective stroke registry during a 10-year period. Main outcome measures were frequency of demographic variables, risk factors, clinical events, neuroimaging data, and outcome in ICH patients with and without diabetes. Variables related to vital status at discharge (alive or dead) in the univariate analysis plus age were studied in 4 logistical regression models. RESULTS: A total of 35 patients (15.3%) had diabetes. The overall in-hospital mortality rate was 54.3% in the diabetic group and 26.3% in the nondiabetic group (P < 0.001). Previous cerebral infarction, altered consciousness, sensory symptoms, cranial nerve palsy, multiple topography of the hematoma, intraventricular hemorrhage, and infectious complications were significantly more frequent in diabetic patients than in nondiabetic patients. The presence of diabetes was a significant predictive variable in the model based on demographic variables and cardiovascular risk factors (odds ratio 2.98 [95% CI 1.37-6.46]) and in the models based on these variables plus clinical variables (5.76 [2.01-16.51]), neuroimaging variables (5.59 [1.87-16.691), and outcome data (6.10 [2.04-18.291). CONCLUSIONS: Diabetes is an independent determinant of death after ICH. ICH in diabetic individuals presents some different clinical features compared with ICH in nondiabetic patients. PMID- 11023149 TI - Epalrestat, an aldose reductase ihibitor, reduces the levels of Nepsilon (carboxymethyl)lysine protein adducts and their precursors in erythrocytes from diabetic patients. AB - OBJECTIVE: To clarify the role of the polyol pathway in the intracellular formation of advanced glycation end products in human tissues, we examined the effects of epalrestat, an aldose reductase inhibitor, on the level of Nepsilon (carboxymethyl)lysine (CML) along with 3-deoxyglucosone (3-DG) and triosephosphates in erythrocytes from diabetic patients. Plasma thiobarbituric acid-reactive substances (TBARS) were also determined as indicators of oxidative stress. RESEARCH DESIGN AND METHODS: Blood samples were collected from 12 nondiabetic volunteers, 38 untreated type 2 diabetic patients, and 16 type 2 diabetic patients who had been treated with 150 mg epalrestat/day. Blood samples were also collected from 14 of the untreated type 2 diabetic patients before and after the administration of epalrestat for 2 months. The amount of erythrocyte CML was determined by a competitive enzyme-linked immunosorbent assay, and 3-DG was measured by high-performance liquid chromatography RESULTS: In diabetic patients not treated with epalrestat, the erythrocyte CML level was significantly elevated above levels seen in nondiabetic individuals (49.9 +/- 5.0 vs. 31.0 +/- 5.2 U/g protein, P < 0.05) and was significantly lower in patients receiving epalrestat (33.1 +/- 3.8 U/g protein, P < 0.05). Similar results were observed with 3-DG. The treatment of patients with epalrestat for 2 months significantly lowered the level of erythrocyte CML (46.2 +/- 5.6 at baseline vs. 34.4 +/- 5.0 U/g protein, P < 0.01) along with erythrocyte 3-DG (P < 0.05), triosephosphates (P < 0.05), fructose (P < 0.05), sorbitol (P < 0.05), and plasma TBARS (P < 0.05) without changes in plasma glucose and HbA(1c) levels. A positive correlation was evident between the erythrocyte CML and sorbitol (r = 0.49, P < 0.01) or fructose (r = 0.40, P < 0.05) levels in diabetic patients. CONCLUSIONS: The results indicate that epalrestat administration lowers CML and associated variables and that polyol metabolites are correlated with CML in the erythrocytes of diabetic patients. The observed results suggest that aldose reductase activity may play a substantial role in the intracellular formation of CML in the mediation of reactive intermediate metabolites and oxidative stress. PMID- 11023148 TI - Variants in the hepatocyte nuclear factor-1alpha and -4alpha genes in Finnish and Chinese subjects with late-onset type 2 diabetes. AB - OBJECTIVE: To determine the role of the hepatocyte nuclear factor (HNF)-1alpha and HNF-4alpha genes in the etiology of late-onset type 2 diabetes in Finnish and Chinese subjects. RESEARCH DESIGN AND METHODS: The whole coding regions of the genes encoding for HNF-1alpha and HNF-4alpha, including approximately 800 bp of the HNF-1alpha promoter, were investigated in 40 Finnish subjects (fasting C peptide 50-570 pmol/l) and 47 Chinese subjects with type 2 diabetes by single strand conformation polymorphism (SSCP) analysis. Frequencies of the variants of these genes were analyzed by restriction fragment-length polymorphism analysis in additional samples of 100 Finnish diabetic patients and 82 Finnish control subjects and in 58 Chinese diabetic patients and 51 Chinese control subjects. RESULTS: No previously reported gene defects were detected, but one novel functionally silent GCC-->GCG variant (nucleotide 73, exon 10) was observed in the HNF-4alpha gene in a Chinese diabetic patient. Interestingly, the Ala98Val substitution of the HNF-1alpha gene occurred at a significantly higher frequency in 140 Finnish diabetic patients compared with 82 control subjects (P = 0.014). The Ala98Val variant was not, however, associated with abnormalities in insulin secretion evaluated by oral and intravenous glucose tolerance tests in subjects with normal (n = 295) or impaired (n = 38) glucose tolerance. CONCLUSIONS: Variants in the HNF-1alpha and HNF-4alpha genes are unlikely to play a major role in the pathogenesis of late-onset type 2 diabetes in Finnish and Chinese subjects. However, the association of the Ala98Val variant of the HNF-1alpha gene with type 2 diabetes in Finnish subjects may indicate a diabetogenic locus close to the HNF-1alpha gene. PMID- 11023150 TI - Skeletal muscle morphology and exercise response in congenital generalized lipodystrophy. AB - OBJECTIVE: Congenital generalized lipodystrophy (CGL) is an autosomal recessive genetic disorder characterized by almost complete absence of adipose tissue, muscular appearance, and severe insulin resistance since birth. We investigated whether insulin resistance in CGL patients is associated with abnormal muscle morphology and whether increased muscularity imparts increased muscle strength and exercise capacity RESEARCH DESIGN AND METHODS: We obtained quadriceps muscle biopsies to study muscle fiber types and capillary density in three African American women (aged 17-20 years) with CGL. We also assessed quadriceps muscle strength, muscle metabolism, and maximal O2 consumption in the patients. RESULTS: Quadriceps muscle biopsies revealed a markedly higher percentage of type II (fast twitch glycolytic) muscle fibers in patients with CGL versus sedentary young women (75-78 vs. 47-57%, respectively). The capillary-to-fiber ratio (2.7-3.0), however, was normal. Cross-sectional areas of type I (slow-twitch oxidative) (1,262-2,685 microm2) and type II (2,304-3,594 microm2) fibers were far below the normal values (3,811-4,310 and 3,115-4,193 microm2, respectively), suggesting muscle hyperplasia but not hypertrophy The quadriceps muscle strength, as measured by Cybex, was below average; the maximal O2 consumption (23-32 ml x kg( 1) x min(-1)) was also below average. 31P nuclear magnetic resonance spectroscopy of the forearm muscles revealed normal pH and metabolic responses to static and dynamic exercises. CONCLUSIONS: We conclude that insulin resistance in patients with CGL is associated with an increased proportion of type II muscle fibers but not reduced capillary density. Increased muscularity in CGL is due to muscle hyperplasia and is not associated with increased muscle strength. PMID- 11023151 TI - Systemic hyperbaric oxygen therapy: lower-extremity wound healing and the diabetic foot. AB - OBJECTIVE: To document peer-reviewed medical publications that have reported on hyperbaric oxygen (HBO) therapy as an adjunct to standard lower-extremity wound care, focusing on publications dealing with the diabetic foot. RESEARCH DESIGN AND METHODS: A review of the medical literature was conducted using MEDLINE. Research articles involving HBO treatment and the diabetic foot were critiqued to identify factors that may have been a source of bias. RESULTS: Of the published reports on human studies, seven involved diabetes-related foot pathology. Five of these studies, two of which were randomized, included a control group that did not receive HBO therapy The controlled diabetic foot studies included an average of 28 subjects in the HBO therapy group (range 10-62) and an average of 16.2 subjects in the non-HBO control group (range 5-33). Most of the published reports have several potential sources of bias, including, but not limited to, inadequate evaluation of comorbid conditions relevant to wound healing, small sample size, and poor documentation of wound size or severity. Four of the seven reports involving the diabetic foot were published by a group of researchers at the University of Milan between 1987 and 1996. CONCLUSIONS: Additional randomized placebo-controlled clinical trials in large diabetic populations would further lend credence to the presumption that HBO therapy improves clinical outcomes. Given the relatively high cost of this treatment modality, perhaps a more acute awareness of the medical literature would reduce the economic burden that HBO therapy imposes on care providers that are financially at risk. PMID- 11023152 TI - A review of the relationship between depression and diabetes in adults: is there a link? AB - OBJECTIVE: To review the support for two hypotheses concerning the interrelationship between depression and diabetes and to identify areas in which more research is needed. RESEARCH DESIGN AND METHODS: A review was conducted using primarily electronic databases. Articles relating to diabetes and depressive symptomatology, depressive disorder, and dysthymic disorder were selected. The study focuses mainly on adults with diabetes. RESULTS: The initial onset of major depressive disorder (MDD) seems to be independent of the onset of type 2 diabetes, but results remain equivocal for type 1 diabetes. However, in both type 1 and type 2 diabetes, diabetes-related psychological and physiological processes may be involved in the higher recurrence and longer duration of MDD and depressive symptomatology CONCLUSIONS: The hypotheses that the initial occurrence of clinically significant depression, MDD, results from either biochemical changes directly due to type 2 diabetes or its treatment or from the psychosocial demands imposed by the illness or its treatment do not seem to be supported. MDD in diabetic individuals represents a multidetermined phenomenon resulting from interactions between biologic and psychosocial factors. This interaction may increase the probability of developing type 2 diabetes in otherwise healthy individuals. PMID- 11023154 TI - 2000 presidential address: American Diabetes Association. PMID- 11023153 TI - Screening for type 2 diabetes. AB - Definitive studies of the effectiveness of screening for type 2 diabetes are currently not available. RCTs would be the best means to assess effectiveness, but several barriers prevent these studies from being conducted. Prospective observational studies may characterize some of the benefits of screening by creating screened and unscreened groups for comparison. The availability of better data systems and health services research techniques will facilitate such comparisons. Unfortunately, the interpretation of the results of such studies is extremely problematic. Several screening tests have been evaluated. Risk assessment questionnaires have generally performed poorly as stand-alone tests. Screening with biochemical tests performs better. Venous and capillary glucose measurements may perform more favorably than urinary glucose or HbA(1c) measurements, and measuring postprandial glucose levels may have advantages over measuring fasting levels. However, performance of all screening tests is dependent on the cutoff point selected. Unfortunately, there are no well-defined and validated cutoff points to define positive tests. A two-stage screening test strategy may assist with a more efficient use of resources, although such approaches have not been rigorously tested. The optimal interval for screening is unknown. Even though periodic, targeted, and opportunistic screening within the existing health care system seems to offer the greatest yield and likelihood of appropriate follow-up and treatment, much of the reported experience with screening appears to be episodic poorly targeted community screening outside of the existing health care system. Statistical models have helped to answer some of the key questions concerning areas in which there is lack of empirical data. Current models need to be refined with new clinical and epidemiological information, such as the UKPDS results (200). In addition, future models need to include better information on the natural history of the preclinical phase of diabetes. Data from ongoing clinical trials of screening and treatment of impaired glucose tolerance, such as the Diabetes Prevention Program, may eventually offer more direct evidence for early detection and treatment of asymptomatic hyperglycemia (201). It will be important to use comprehensive cardiovascular disease modules that assess the conjoint influence of glucose and cardiovascular risk factor reduction, information on QOL, and refined economic evaluations using common outcome measures (cost per life-year or QALY gained) (11,178,202-204). Such studies should consider all of the costs associated with a comprehensive screening program, including, at a minimum, the direct costs of screening, diagnostic testing, and care for patients with diabetes detected through screening. Finally, combinations of screening tests and different screening intervals should be evaluated within economic studies to allow selection of the optimal approach within the financial and resource limitations of the health care system. PMID- 11023155 TI - Obesity and diabetes. PMID- 11023156 TI - Improvement of insulin sensitivity after adrenalectomy in patients with pheochromocytoma. PMID- 11023157 TI - Association of autoimmune pancreatitis and type 1 diabetes: autoimmune exocrinopathy and endocrinopathy of the pancreas. PMID- 11023158 TI - Use of sibutramine hydrochloride monohydrate in the treatment of the painful peripheral neuropathy of diabetes. PMID- 11023159 TI - Insulin sensitivity indexes calculated from oral glucose tolerance test data. PMID- 11023160 TI - Regarding depressive symptoms and metabolic control in African-Americans with type 2 diabetes. PMID- 11023161 TI - HbA1c determination and predicting the clinical status of diabetic patients. PMID- 11023162 TI - Developmental outcome of extremely preterm infants. AB - Institution-based studies from perinatal centers are reporting encouraging survival and developmental outcome in extremely preterm infants, but population based studies of all such births within a geographical-defined region are necessary to examine the impact of perinatal-neonatal care on the entire community. We have reported that their perinatal mortality and severe disability rate were significantly lower in those born within perinatal centers compared with those born elsewhere. Promotion of "in utero transfer" is associated with a significant improvement in their survival and developmental outcome. Postnatal surfactant therapy, which began a decade ago, saw a doubling of the survival rate for infants born as early as 24-26 weeks, while their severe disability rate remains below 10%. Although surfactant and mechanical ventilation are expensive therapies, the resulting improvement in outcome was responsible for a reduction in the cost per additional quality-adjusted life-year gained. PMID- 11023163 TI - Recent cocaine use is not associated with fetal acidemia or other manifestations of intrapartum fetal distress. AB - The objective of this paper is to evaluate the impact of recent cocaine use on umbilical cord blood gas values in cocaine-dependent pregnant women who received formal prenatal care. Ninety-two cocaine-dependent pregnant women receiving comprehensive prenatal care were divided into two groups, with patients in Group A (n = 35) testing positive for cocaine metabolites at the time of delivery and Group B (n = 57) testing negative. One hundred and three patients with no history of drug or alcohol dependence served as the control group (Group C). Umbilical cord blood gases were obtained at all deliveries. Additional variables included 1 and 5-min Apgar scores, meconium staining of the amniotic fluid, route of delivery, premature rupture of the membranes, and length of nursery stay. There were no statistically significant differences between groups in either umbilical artery pH, pO2, pCO2, bicarbonate, or base excess. Similarly, there was no difference in meconium staining of the amniotic fluid, depressed Apgar scores, cesarean delivery, or neonatal length of stay. Our data do not support an association between recent cocaine use and fetal hypoxemia or acidemia, depressed 5-min Apgar scores, meconium staining of the amniotic fluid, or cesarean delivery in cocaine-dependent pregnant women enrolled in prenatal care. PMID- 11023164 TI - Persistent pulmonary hypertension after maternal naproxen ingestion in a term newborn: a case report. AB - Constricting effect of indomethacin on the ductus arteriosus of the fetus is well known. The fetal effects of other nonsteroid anti-inflammatory drugs (NSAIDs) like naproxen are not well reported. We report here a case of a 3,790-g term neonate who developed persistent pulmonary hypertension after birth with a closed ductus arteriosus. The mother admitted to taking naproxen sodium immediately prior to the birth of the infant. The course of illness was progressively better on conservative management. Like indomethacin, other NSAIDs can also cause premature closure of fetal ductus arteriosus, pulmonary hypertension, and life threatening problems to the neonate. Patient education regarding over-the-counter pain medication during pregnancy should be emphasized. PMID- 11023165 TI - Does the antenatal detection of small-for-gestational-age babies influence their two-year outcomes? AB - The aim of this paper is to determine whether antenatal detection of small-for gestational-age (SGA) babies influences 2-year outcomes. All low-birth-weight (<2,500g) infants born in South-EastThames region, England from September 1, 1992 to August 31, 1993 were identified at birth. Antenatal "suspicion" and ultrasound assessment confirming growth restriction was categorized as "detection" of SGA. Postnatally, infants were classified as SGA if they had a birth weight for given gestation below the 10th centile. At 2 years, those below 32 weeks' gestation and a random 25% sample of infants of 32 weeks' gestation or more underwent pediatric assessments. Of 49,787 births, 3,456 (6.9%) were of low birth weight. One thousand four hundred and fifty one (42.5%) were SGA, of whom 611 (42%) were detected antenatally by ultrasound scan. At 2 years, 1,008 (75.8%) of 1,358 expected infants were assessed, 379 (37.6%) were SGA at birth, and 188 (49.6%) were confirmed antenatally. Although undetected infants had higher mean birth weights and gestational ages, they had a higher proportion of perinatal deaths (12.6 vs. 6.4%, RR 1.96: CI 1.32-2.86) than detected infants. At 2 years, detected SGA infants had smaller head circumferences (p = 0.026), a higher prevalence of febrile convulsions (8.0 vs. 3.1 %: p = 0.040) and lower scores on the locomotor (DQA) scale of Griffith's developmental test (p = 0.021) compared with undetected SGA infants. Despite detected SGA fetuses having lower weights and gestation at birth than undetected fetuses, they had significantly lower mortality without a parallel increase in severe 2-year neuro-developmental, clinical, or growth morbidity. PMID- 11023166 TI - Bacterial vaginosis and cervical dilation and effacement at 24-29 weeks' gestation. AB - The purpose of this study was to investigate the association between bacterial vaginosis (BV) and cervical dilation and effacement, as measures of impending preterm delivery. The Pregnancy, Infection, and Nutrition Study collected genital tract specimens and documented cervical change from 807 eligible women between 24 and 29 weeks' gestation. BV was assessed with Nugent-scored vaginal smears, and analyzed in relation to cervical measurements. At 24-29 weeks' gestation, <7% of women had a dilated cervix, 31% had a cervix < or =2 cm, and 17.3% had BV. Unadjusted analyses found no associations between BV and cervical measurements. Adjusted logistic regression suggested an association between BV and cervical effacement among women with a sexually transmitted disease (STD) earlier in pregnancy (odds ratio = 1.9, 95% CI 0.8-4.3). Stratified analyses for BV/dilation also suggested interaction with STDs. Overall, BV was not association with cervical dilation or effacement at 24-29 weeks' gestation. PMID- 11023167 TI - Asymmetrical septal hypertrophy in newborn infants of diabetic mothers. AB - The objective of this paper is to determine the frequency and outcome of asymmetrical septal hypertrophy (ASH) in large-for-gestational-age infants (LGA) born to diabetic (DM) and nondiabetic mothers (NDM), and to establish the relationship between ASH and maternal diabetes control. A comparative study was design to assess ASH in infants born to DM and NDM. The study was conducted in the Departments of Neonatology and Pediatric Cardiology of the "Hospital de Gineco-Pediatria 48", Instituto Mexicano del Seguro Social from January to December 1997. Eighty-five full-term infants of DM (group A) and 85 LGA infants of NDM (group B) were included. As a control group (group C), we studied 85 healthy, full-term infants. In all cases a Doppler echocardiogram was obtained in the first 48 h after birth, and for the ASH infants, at 2 and 4 months. Chest X ray, electrocardiogram, and laboratory tests were performed as complementary studies. ASH was present in 38.8% of LGA infants of DM and in 7.1% of NDM. The difference was significant (p < 0.01). Interventricular septum (IVS) and IVS/ posterior wall of left ventricle ratio were significantly different between groups A and B with C. There was no correlation between Hb A1 level and the presence of ASH in group A. ASH is a common finding in infants of DM. We could not find a relationship between the degree of metabolic control during pregnancy and the incidence and severity of ASH. PMID- 11023168 TI - Assessment of therapy for arterial hypotension in critically ill preterm infants. AB - The aim of this paper is to assess the efficacy of albumin and dopamine compared with albumin and dobutamine in treating hypotension in preterm newborn infants (PNI). A randomized, open-label, prospective, cross-over study, was designed on 66 PNI whose weights were between 1,000 to 1,500 g, and persistent hypotension, defined as a mean arterial pressure (MAP) of < 30 mmHg. Infants were randomly allocated to two groups and received a 5% albumin infusion at a dosage of 20 mL/kg, in 30 min. Thereafter, one group received dopamine and the other dobutamine at doses of 5 microg/kg/min. If there was not an increase in MAP values > 30 mmHg, the infusions were increased every 20 min by 2.5 microg/kg/min, up to a maximum of 10 microg/kg/min. Treatment failure was considered when there was no pressure response within 2 hr after the infusion started; then patients were changed to the other catecholamine. Statistical analysis was done with student's t-test, x2, and Fisher's exact probability test. There were no differences between groups in initial features. Overall, MAP was normalized with dopamine in 29 of 33 infants and with dobutamine in 25 of 33 infants (p > 0.05). The initial dosage of 5 microg/kg/min, was adequate in 22 infants treated with dopamine and in 13 treated with dobutamine (p < 0.05). The change from dopamine to dobutamine was successful in three out of four patients, while changing from dobutamine to dopamine was adequate in seven out of eight patients. Dopamine is recognized as the drug of choice to treat hypotension in PNI. Since our results showed only small differences in responses, it is proposed that dobutamine is also as efficacious and useful as dopamine. PMID- 11023169 TI - Maternal complications with placenta previa. AB - The purpose of this study is to describe the maternal complications of placenta previa. A population-based retrospective cohort study including all women delivered in the province of Nova Scotia, Canada from 1988 to 1995 was performed. Patient information was obtained from the Nova Scotia Atlee Perinatal Database and maternal complications were described for all women undergoing cesarean delivery. Prognostic factors for the risk of hysterectomy in woman with placenta previa were analyzed by multiple logistic regression. During the 8-year period, 308 cases of placenta previa were identified in 93,996 deliveries (0.33%). Maternal complications included hysterectomy [relative risk (RR) = 33.26], antepartum bleeding (RR = 9.81), intrapartum (RR = 2.48), and postpartum (RR = 1.86) hemorrhages, as well as blood transfusion (RR = 10.05), septicemia (RR = 5.55), and thrombophlebitis (RR = 4.85). Risk factors for need of hysterectomy in women with placenta previa include the presence of placenta accreta and previous cesarean delivery. PMID- 11023170 TI - Neutrophil chemotaxis and random migration in preterm and term infants with sepsis. AB - The aim of this study was to evaluate neutrophil chemotaxis and random migration in healthy newborn infants and septic neonates with similar gestational and postnatal age. Possible relationships between chemotactic activity, random migration, causative microorganisms, and clinical course of septic infants were also investigated. The neutrophil chemotaxis and random migration was evaluated in 24 healthy newborn babies and 34 septic neonates and 20 healthy adults by modified Boyden technique. The mean neutrophil chemotaxis of healthy preterm-term infants and adults were similar (66.6 +/- 18.9, 64.4 +/- 19.9, and 74.7 +/- 17 microm, respectively). The mean neutrophil random migration of healthy term infants was not different than that of adults. But the mean neutrophil random migration of healthy preterm infants was lower than that of adults (36.9 +/- 13.7 and 43.5 +/- 1 1.8 microm, respectively) (p = 0.03). The mean neutrophil chemotaxis and random migration septic term infants were not different from the value of healthy term infants (p > 0.05). Although the mean random migration of septic and preterm infants were similar (p > 0.05), the mean neutrophil chemotaxis of septic preterm infants was lower than the value of healthy preterm infants (p = 0.04). Not only mean neutrophil chemotaxis of septic preterm and term infants were significantly lower than that of adults (p = 0.002 and p = 0.006, respectively), but also neutrophil random migration of septic preterm and term infants were significantly lower than that of adults (p = 0.001 and p = 0.005, respectively). There was no relationship between the nature of causative microorganism and neutrophil random migration or chemotactic activity. Polymorphonuclear leukocytes chemotaxis was significantly lower in preterm with sepsis compared with healthy preterm-term infants and adults. These findings may indicate deterioration in neutrophil functions in premature babies under stress but more detailed studies with larger groups are needed. PMID- 11023171 TI - Introduction: pediatric epilepsy surgery in 2000. PMID- 11023172 TI - Consequences of epilepsy in the developing brain: implications for surgical management. AB - The developing brain is highly susceptible to seizures, as demonstrated by both human and animal studies. Until recently, the brain has been considered to be relatively resistant to damage induced by seizures early in life. Accumulating evidence in animal models now suggests that early seizures can cause structural and physiologic changes in developing neural circuits that result in permanent alterations in the balance between neuronal excitation and inhibition, deficits in cognitive function, and increased susceptibility to additional seizures. The disruption of normal neuronal activity by seizures can affect multiple developmental processes, resulting in these long-lasting changes. These data should be considered in the clinical approach to children with intractable epilepsy and suggest that early intervention may avoid some of these long-term neurologic deficits. PMID- 11023173 TI - Intractable pediatric epilepsy: presurgical evaluation. AB - Epilepsy surgery in children requires a multidisciplinary approach. This section examines the role of scalp EEG, video-EEG monitoring, and intracranial EEG in the presurgical evaluation. Concepts central to understanding the basis for surgical treatment, such as the epileptogenic zone, the irritative zone, and the epileptogenic lesion, are discussed. An illustrative case then demonstrates application of the process in clinical practice. Neuroimaging and neuropsychological issues are not discussed herein; rather they are addressed elsewhere in this issue. PMID- 11023174 TI - Pediatric epilepsy surgery: neuroimaging, neuropsychology, and anticonvulsants. AB - Neuroimaging and the neuropsychological evaluation are important components of the presurgical evaluation for epilepsy surgery. Advances in neuroimaging over the last decade, to a large part, underlie improvements in pediatric epilepsy surgery outcomes. The neuropsychological evaluation plays an important role in the evaluation of the older child and adolescent, particularly in the evaluation of mesial temporal sclerosis. However, its role in the young child being considered for surgery remains to be defined. This section reviews the definition of medical intractability, issues related to medication withdrawal during video EEG monitoring, recent neuroimaging advances, and the neuropsychological evaluation. PMID- 11023175 TI - Recent advances in candidate selection for pediatric epilepsy surgery. AB - Many patients who were once considered inoperable are candidates for a wide variety of surgical procedures. This trend is particularly striking at pediatric centers that serve patients with diverse forms of epilepsy that compromise development. Newer diagnostic techniques, including structural and functional imaging, EEG monitoring, and surgical technology, can provide a complete representation of the etiology and expression of the epileptic focus, including accurate cortical localization. This review focuses on several forms of epilepsy that, until recently, were not regarded to be surgically amenable. The adverse seizure and neurobehavioral prognosis in patients with these disorders provides the justification to search for innovative treatments. With more sophisticated tools for preoperative evaluation and an expanding knowledge base, it is likely that the number of surgical candidates will continue to increase in the future. PMID- 11023176 TI - Epilepsy surgery in infancy. AB - Surgery for intractable epilepsy is being offered at progressively younger ages, including infancy. The most common causes of catastrophic epilepsy in very young surgical candidates are focal malformations of cortical development and low-grade tumors. Additional causes include Sturge-Weber syndrome, epidermal nevus syndrome, hemimegalencephaly, and prenatal or perinatal infarction. Many infants manifest with focal seizures, whereas some patients have infantile spasms in the setting of a focal epileptogenic lesion. Video electroencepholography, magnetic resonance imaging, and positron emission tomography are critical investigations to explore surgical options. In small series, the percentage of infants free of seizures after surgery was in the range of 60%. This is similar to that seen after epilepsy surgery in older children, adolescents, and adults. However, larger series with long-term follow up will be important. Furthermore, the extensive procedures required in infants for removal of the epileptogenic developmental lesions entail some risk, and should not be offered in the absence of severe epilepsy. Most infant candidates for epilepsy surgery have significant developmental delay. Few data are available, but anecdotal experience suggests that surgical relief of catastrophic epilepsy may result in resumption of developmental progression. For each infant, the timing of surgery must be carefully considered based on full assessment of the relative risks and benefits, derived from a detailed presurgical evaluation. PMID- 11023177 TI - Epilepsy surgery in children with developmental disabilities. AB - Surgery for treatment of medically uncontrolled epilepsy in children is now widely accepted with reported outcomes similar to those in adults. Epilepsy is reported in 8.8% to 32% of children with mental retardation (MR) and in up to half of children with severe retardation. There has been concern that patients with low IQ will experience unsatisfactory outcomes from epilepsy surgery and not achieve good seizure control. It is appropriate to reassess the prior bias against resective epilepsy surgery in children with MR in view of the changing criteria for potential candidacy for epilepsy surgery in infants and young children. There are three prerequisites for epilepsy surgery: (1) the epilepsy must be medically intractable; (2) the surgery must be feasible, that is, the epileptogenic zone can identified and safely resected; and (3) there is high likelihood of a satisfactory outcome as regards both the epilepsy and the patient's functional status. Patients with MR may have diffuse cerebral dysfunction and diffuse or multifocal epileptogenic regions. Appropriate patient selection is made possible through use of current technology that allows identification of lesions or areas of cerebral dysgenesis, aiding in identification of localized areas of epileptogenesis. Results from various series of patients with MR who have undergone resective surgery for epilepsy have shown that with careful presurgical evaluations, outcomes are similar between patients with normal IQ scores and those with low scores. Surgical protocols specifically for patients with MR and intractable epilepsy are required, including careful definition of desired outcomes. PMID- 11023178 TI - Epilepsy surgery in children, with special attention to focal cortical resections. AB - Epilepsy surgery is an important alternative for children and adolescents with medically intractable epilepsy. Advances in imaging have allowed the visualization of epileptogenic regions, including cortical dysplasia. The presurgical evaluation will likely become less invasive as imaging technology improves. Long-term development is a critical outcome measure, in addition to seizure control. The long-term prognosis after epilepsy surgery may be related to factors other than seizure control, including the timing of the intervention and the cause of the epilepsy. It may be useful to independently analyze children based on cause, and further studies examining the long-term outcome should become feasible as surgical experience grows. PMID- 11023179 TI - Vagal nerve stimulation in children. AB - Vagal nerve stimulation is a new therapeutic option for patients with medically refractory epilepsy. The FDA approved the NeuroCybernetic Prosthesis (NCP) in July 1997 for use in adults and adolescents over the age of 12 years with medically refractory epilepsy. Most of the patients in the initial pilot studies and subsequent extended longitudinal and randomized controlled studies were adults. There were small numbers of children who received the NCP system. However, these were not part of controlled studies. As the system has had greater exposure in the United States and Europe, there are more children who are receiving vagal nerve stimulation (VNS). Initial data from open-label, uncontrolled studies suggest that VNS does have some efficacy and safety for those children with refractory epilepsy who have not responded to appropriate trials of antiepileptic drugs. The questions to be asked and answered are as follows: (1) When is a child medically refractory? (2) What are the criteria for selection for VNS? (3) Which seizure types or syndromes will benefit most from the treatment? and (4) What are the most effective and safe stimulation parameters, and do these vary depending on the seizure type? PMID- 11023180 TI - A pictoral guide to pediatric epilepsy surgery. AB - This article contains a discussion and detailed pictorial account of diagnostic and neurosurgical therapeutic procedures used to help pediatric patients who have medically intractable epilepsy. The operations discussed are placement of sphenoidal electrodes, strips and grids, temporal and extratemporal resections, corpus callosotomy, and hemispherectomy. PMID- 11023181 TI - Host-microbe interactions in the pathogenesis of invasive group A streptococcal infections. PMID- 11023182 TI - Diagnostic particle agglutination using ultrasound: a new technology to rejuvenate old microbiological methods. AB - Microbial antigen in clinical specimens can be detected rapidly by commercial test-card latex agglutination, but poor sensitivity is a potential difficulty. Antigen detection by immuno-agglutination of coated latex micro-particles can be enhanced in comparison with the conventional test-card method in both rate and sensitivity by the application of a non-cavitating ultrasonic standing wave. Antibody-coated micro-particles suspended in the acoustic field are subjected to physical forces that promote the formation of agglutinates by increasing particle particle contact. This report reviews the application of ultrasound to immuno agglutination testing with several commercial antibody-coated diagnostic micro particles. This technique is more sensitive than commercial card-based agglutination tests by a factor of up to 500 for fungal cell-wall antigen, 64 for bacterial polysaccharide and 16 for viral antigen (in buffer). The detection sensitivity of meningococcal capsular polysaccharide in patient serum or CSF has been increased to a stage where serotyping by ultrasound-enhanced agglutination is comparable to that achievable with the PCR, but is available more rapidly. Serum antigen concentration as measured by ultrasonic agglutination has prognostic value. Increasing the sensitivity of antigen detection by increasing the acoustic forces that act on suspended particles is considered. Employing turbidimetry to measure agglutination as part of an integrated ultrasonic system would enable the turnover of large numbers of specimens. Ultrasound-enhanced latex agglutination offers a rapid, economical alternative to molecular diagnostic methods and may be useful in situations where microbiological and molecular methods are impracticable. PMID- 11023183 TI - Enumeration of Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans in subgingival plaque samples by a quantitative-competitive PCR method. AB - Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans are believed to play an important role in adult periodontitis, but the significance of their relative numbers and progress of the disease is still unclear. Traditional quantitative methods are generally time consuming and inaccurate. The aim of this study was to develop a sensitive, quantitative PCR technique that would be useful for enumerating P. gingivalis, Pr. intermedia and A. actinomycetemcomitans in subgingival plaque samples from subjects with adult periodontitis. Primers to the following genes were employed: the fimbrial gene (fimA) of P. gingivalis, the 16S rRNA gene of Pr. intermedia and the leukotoxin-A (lktA) gene of A. actinomycetemcomitans. Competitive templates were constructed either by sequence deletion between primer binding sites or by annealing of the primer binding sites to an appropriate DNA core so as to yield products of a different size from that obtained with the target template. Coamplification of target and competitive templates yielded products of expected size and non-specific recognition by the primers was not found. The sensitivity of the designed primers was 100 cells of P. gingivalis, 100 cells of Pr. intermedia and 10 cells of A. actinomycetemcomitans. The three species were found in subgingival plaque samples collected from both healthy and diseased sites by the quantitative-competitive (QC)-PCR method and the technique was more sensitive than cultural methods. For determining the proportions of each of the three periodontopathogens, the total number of bacteria in the samples was enumerated by quantitative-PCR with 16S rRNA universal primers (27f and 342r). The findings indicate that QC-PCR is a useful method for enumerating bacteria in clinical oral specimens and the technique could play a role in the investigation of disease progression. PMID- 11023184 TI - Preferential adherence of cable-piliated burkholderia cepacia to respiratory epithelia of CF knockout mice and human cystic fibrosis lung explants. AB - The Burkholderia cepacia complex consists of at least five well-documented bacterial genomovars, each of which has been isolated from the sputum of different patients with cystic fibrosis (CF). Although the world-wide prevalence of this opportunist pathogen in CF patients is low (1-3%), 'epidemic' clusters occur in geographically isolated regions. Prevalence in some of these clusters is as high as 30-40%. The majority of CF B. cepacia isolates belong to genomovar III, but the relationship between genomovar and virulence has not yet been defined. Because the initial stage of infection involves bacterial binding to host tissues, the present study investigated differences in the binding of representative isolates of all five genomovars to fixed nasal sections of UNC cftr (-/-) and (+/+) mice and to human lung explants, biopsy and autopsy tissue of CF and non-CF patients. Binding was highest for isolates of genomovar III, subgroup RAPD type 2, but only if the isolates expressed the cable pili phenotype. Antibodies to the 22-kDa adhesin of cable pili virtually abolished binding. Binding occurred only to cftr (-/-) nasal sections or to CF lung sections and was negligible in cftr (+/+) or human non-CF, histologically normal lung sections. Unlike normal epithelia, the hyperplastic epithelia of CF bronchioles were enriched in cytokeratin 13, a 55-kDa protein that has previously been shown to act as a receptor in vitro for cable-piliated B. cepacia. These findings may help to explain the high transmissibility ofCbl-positive, genomovar III strains of B. cepacia among CF patients. PMID- 11023185 TI - Cloning, sequencing and characterisation of a Listeria monocytogenes gene encoding a fibronectin-binding protein. AB - Listeria monocytogenes is a gram-positive, non-sporulating food-borne pathogen of man and animals that is able to invade many eukaryotic cells. L. monocytogenes possesses several proteins that bind fibronectin. In this study, an L. monocytogenes DNA library in pUC19 was screened with fibronectin and a gene encoding a 24.6-kDa fibronectin-binding protein (Fbp) was isolated and sequenced. Transcripts of the fbp gene were found in wild-type, in deltaprfA, and PrfA-S183A strains, despite the presence of a 'PrfA-like' box around its ribosome-binding site. The fbp gene was found to be present in all tested isolates of the species L. monocytogenes and a homologous DNA fragment was amplified in L. welshimeri. No homologies between the fbp gene and its translation product with any other DNA or proteins deposited in databanks were found. Restriction endonuclease-PCR (RE-PCR) showed that the fbp gene displays a degree of allelic variation among isolates of L. monocytogenes, whereas the corresponding amplified fragment of L. welshimeri seems to be monomorphic among isolates of this species. RE-PCR with Hha I, Dde I or Taq I produced DNA banding profiles specific for each of these two species, allowing their identification. PMID- 11023186 TI - Infection of human enterocyte-like cells with rotavirus enhances invasiveness of Yersinia enterocolitica and Y. pseudotuberculosis. AB - Mixed infection with rotavirus and either Yersinia enterocolitica or Y. pseudotuberculosis was analysed in Caco-2 cells, an enterocyte-like cell line highly susceptible to these pathogens. Results showed an increase of bacterial adhesion and internalisation in rotavirus-infected cells. Increased internalisation was also seen with Escherichia coli strain HB101 (pRI203), harbouring the inv gene from Y. pseudotuberculosis, which is involved in the invasion process of host cells. In contrast, the superinfection with bacteria of Caco-2 cells pre-infected with rotavirus resulted in decreased viral antigen synthesis. Transmission electron microscopy confirmed the dual infection of enterocytes. These data suggest that rotavirus infection enhances the early interaction between host cell surfaces and enteroinvasive Yersinia spp. PMID- 11023187 TI - Effects of interaction between Escherichia coli verotoxin and lipopolysaccharide on cytokine induction and lethality in mice. AB - In Escherichia coli 0157 infections, verotoxins (VT) play a critical role in causing the disease, although other factors such as lipopolysaccharide (LPS) and inflammatory cytokines may affect the progression and course of the disease. The present study examined the roles of VT and LPS in induction of serum cytokines and lethality in mice. LD50 of VT2 (13 ng) was c. 10(4)-fold smaller than that of LPS (400 microg). Although the lethal toxicity of these toxins was examined in several experimental conditions, such as VT2 (5, 10, 20, 40 ng/mouse) alone or in combination with LPS (100 microg/mouse) at various times (-2 days to +2 days), no evidence of synergy was observed. VT2 did not augment LPS-induced tumour necrosis factor-alpha (TNF-alpha) or interleukin-6 production, and conversely suppressed TNF-alpha production when it was injected 2 days before LPS challenge. The data failed to indicate either synergic or additive effects of VT and LPS on cytokine production or lethality in mice. In contrast, antagonistic interactions were clearly observed in cytokine production in certain conditions. The results suggested that these toxins may be co-operatively involvedin the pathology of VT related diseases, but not through synergic interactions. PMID- 11023188 TI - False-negative serology in patients with neuroborreliosis and the value of employing of different borrelial strains in serological assays. AB - The risk of obtaining false-negative results in serological assays in serum and CSF specimens with only one strain of Borrelia burgdorferi sensu lato as antigen was investigated in 79 patients with neuroborreliosis with specimens obtained at initial presentation. Serum antibodies were assessed by immunoblotting; the criteria of Hauser et al. were used to evaluate the test. The intrathecal synthesis of borrelial-specific IgM and IgG antibodies was examined by enzyme immunoassay (EIA). Strains of B. burgdorferi sensu stricto (BbZ160), B. garinii (Bbii50) and B. afzelii (PKO) served as sources of antigen in both assays. All patients produced either a positive IgM or IgG test in serum with at least one strain of B. burgdorferi sensu lato. Reactivity of IgM or IgG antibodies, or both, with antigens of all three strains was demonstrated in 67 (85%) of 79 sera. The correlation of results of immunoblotting with different strains was significantly better for IgG (85%) than for IgM antibodies (54%). The variability of positive IgM reactions in 18 specimens was mainly due to the fact that the antibodies were directed to the relevant variable outer-surface protein C (p23). Intrathecal synthesis of IgG antibodies was demonstrated in 58 patients (81%) of 72 and of IgM antibodies in 25 of 58 patients. No patient had isolated intrathecal synthesis of IgM antibodies. The majority of CSF samples (56 of 58) were assessed as IgG antibody-positive, independent of the borrelial strain used as antigen in EIA, whereas only 10 of 25 IgM antibody-positive CSF specimens reacted with all three strains. All patients in the study had intrathecal antibody synthesis demonstrable at 6-week follow-up. From this study it is concluded that there is a small, but real, risk of false-negative serological findings at the time of initial clinical presentation in patients with typical symptoms of neuroborreliosis. In these patients a negative serological result with one strain should prompt the repetition of the test with other strains of B. burgdorferi sensu lato. PMID- 11023189 TI - Borreliacidal activity of early Lyme disease sera against complement-resistant Borrelia afzelii FEM1 wild-type and an OspC-lacking FEM1 variant. AB - Sera obtained from 14 Lyme borreliosis patients at early stages (stages I and II) of the disease were examined for their borreliacidal properties against Borrelia afzelii isolate FEM1 by use of a growth inhibition assay. Five of 14 immune sera exhibited borreliacidal activity against isolate FEM1. Heat-inactivated immune sera failed to kill the spirochaetes. Immunoblotting experiments with outer membrane preparations showed that OspC and 11 additional proteins of 14.0, 16.0, 17.7, 19.3, 21.7, 27.5, 32.7, 40.7, 48.9, 51.3 and 53.6 kDa were recognised by borreliacidal immune sera. To analyse the borreliacidal properties of anti-OspC antibodies, two sera (EM4 and EM5), which beside antibodies against a 51.3-kDa protein contained exclusively anti-OspC antibodies, were further investigated by comparative analysis with a FEM1 wild-type and a FEM1 variant lacking OspC in a growth inhibition assay. Only FEM1 wild-type and not variant FEM1OspC(-) was killed by immune sera EM4 and EM5. Complement-dependent killing of FEM1 wild-type was mediated by formation of the terminal complement complex that was found to be attached directly to the outer membrane as confirmed by immuno-electron microscopy. No complement deposition was observed on the surface of variant FEM1OspC(-) after incubation with immune sera EM4 and EM5, thereby suggesting that only anti-OspC antibodies in these two immune sera were responsible for borreliacidal activity. These results provide direct evidence that anti-OspC antibodies, once developed during the immune response, are of critical importance for efficient killing of borreliae in the early phase of infection. PMID- 11023190 TI - Distribution and in-vitro transfer of tetracycline resistance determinants in clinical and aquatic Acinetobacter strains. AB - Following characterisation by phenotypic tests and amplified ribosomal DNA restriction analysis (ARDRA), 50 tetracycline-resistant (MIC > or = 16 mg/L) Acinetobacter strains from clinical (n = 35) and aquatic (n = 15) samples were analysed by PCR for tetracycline resistance (Tet) determinants of classes A-E. All the clinical strains were A. baumannii; most (33 of 35) had Tet A (n = 16) or B (n = 17) determinants, and only two did not yield amplicons with primers for any of the five tetracycline resistance determinants. The aquatic strains belonged to genomic species other than A. baumannii, and most (12 of 15) did not contain determinants Tet A-E. Strains negative for Tet A-E were also negative for Tet G and M; further analysis of two aquatic strains with specific primers for Tet O and Tet Y and degenerate primers for Tet M-S-O-P(B)-Q also showed negative results. Transfer of tetracycline resistance was tested for 20 strains with three aquatic Acinetobacter strains and Escherichia coli K-12 as recipients. Transfer of resistance was demonstrated between aquatic strains from distinct ecological niches, but not from clinical to aquatic strains, nor from any Acinetobacter strain to E. coli K-12. Most transconjugants acquired multiple relatively small plasmids (<36 kb). Transfer did not occur when DNA from the donor strains was added to the recipient cultures and was not affected by deoxyribonuclease I, suggesting a conjugative mechanism. It is concluded that Tet A and B are widespread among tetracycline-resistant A. baumannii strains of clinical origin, but unknown genetic determinants are responsible for most tetracycline resistance among aquatic Acinetobacter spp. These differences, together with the inability of clinical strains to transfer tetracycline resistance in vitro to aquatic strains, contra-indicate any important flow of tetracycline resistance genes between clinical and aquatic acinetobacter populations. PMID- 11023191 TI - Characterisation of a new isolate of Mycobacterium shimoidei from Finland. AB - This report describes the first isolation of Mycobacterium shimoidei in Finland from a sputum specimen obtained from an elderly female patient. M. shimoidei, a potential lung pathogen, is difficult to identify by routine methods and only a few cases have been reported. The present study demonstrated that M. shimoidei has a characteristic pattern for fatty acids and alcohols in gas liquid chromatography. This chromatogram and the pattern of mycolic acids on thin-layer chromatography allow it to be distinguished routinely. The unique sequence of the 16S rRNA gene and the 16S-23S rDNA spacer region allows identification by molecular methods. PMID- 11023192 TI - A comparison of Columbia blood agar with or without oxolinic acid/metronidazole for the isolation of beta-haemolytic streptococci from throat swabs. PHLS (Midlands) Bacterial Methods Evaluation Group. PMID- 11023193 TI - P21 gene expression as an indicator for the activity of adenovirus-p53 gene therapy in non-small cell lung cancer patients. AB - Alterations in the tumor suppressor gene p53 lead to impaired cell cycle control, allowing for the development and growth of tumors. To restore a loss of p53 function, we performed a phase I study of intratumoral gene therapy with adenovirus expressing wild-type p53 in patients with non-small cell lung cancer carrying mutations in the p53 gene. Furthermore, in a phase II study, gene therapy was complemented with simultaneous cisplatin/vinorelbine treatment. Biopsies were obtained from all treated patients before and 24-48 hours after gene therapy to study changes in the expression of p53 target genes. We report here that in most of the cases, the target gene p21 was up-regulated, especially when injection of higher doses of p53-expressing adenovirus was combined with simultaneous chemotherapy, whereas Pig3, previously reported to be highly up regulated by p53, generally did not show a clear increase. Interestingly, a clear p21 gene response was observed only in tumors showing stabilization or regression. We conclude that p21 appears to be up-regulated after adenovirus mediated p53 gene transfer and is the most sensitive marker tested for biological response to gene therapy in the small cohort of non-small cell lung cancers that were studied. PMID- 11023194 TI - Induction of melanoma-associated antigen systemic immunity upon intratumoral delivery of interferon-gamma retroviral vector in melanoma patients. AB - A total of 17 patients with metastatic melanoma were treated with intratumoral interferon-gamma (IFN-gamma) retroviral vector in a phase I clinical trial. A cycle of treatment consisted of five daily injections every 2 weeks. Patients were divided into two treatment arms that involved a single course (one cycle) of treatment (group I; n = 9) and multiple cycles (six cycles) of treatment (group II; n = 8). Patients received intratumoral injections of IFN-gamma (10(7) plaque forming units/mL administered at 0.3, 0.5, and 1.0 mL per cohort of patients). All patients receiving multiple injections either maintained stable disease (n = 5) or achieved a partial or complete response (n = 3) of the injected lesion, whereas in patients receiving a single cycle of treatment, only one of nine patients had a response. Patients were assessed for immunoglobulin G antibody (Ab) responses to the melanoma-associated antigens (MAA) tyrosinase, gp100, TRP 2, and MAGE-A1 by affinity enzyme-linked immunosorbent assay. Anti-MAGE-A1 and tyrosinase Ab were significantly elevated from baseline (day 0) to week 16 during treatment (P = .005; P = .002, respectively) in patients who received multiple injections. Patients undergoing treatment who had a clinical response (stable disease or better) also had significantly more elevated Ab responses to a greater number of MAA (P = .0004). The induction of systemic Ab responses to multiple MAA also correlated with systemic clinical responses. These studies suggest that multiple anti-MAA Ab responses are associated with clinical responses to IFN gamma retroviral treatment and may be used as surrogate response markers. PMID- 11023195 TI - Gene-modified spontaneous Epstein-Barr virus-transformed lymphoblastoid cell lines as autologous cancer vaccines: mutated p21 ras oncogene as a model. AB - The transfer of genes coding for tumor antigens (Ags) into Ag-presenting cells is a promising strategy to develop cancer vaccines for patients not limited by major histocompatibility complex restriction. To test whether autologous lymphoblastoid cell lines (LCL) can be used as an alternative to dendritic cells for Ag presentation, we established LCL by spontaneous growth in cyclosporin-containing medium in vitro (SP-LCL). SP-LCL, which have an advantage over B95-8-transfected LCL in that they carry no risk of introducing a new infectious agent when used for vaccination, served as a permanent source for transfection with an episomal Epstein-Barr virus-based expression vector encoding the Ki-ras p21 oncogene carrying a point mutation at codon 12 (muRas) as a model tumor Ag. The ability of muRas-transfected SP-LCL (muRas-LCL) to induce immunoreactivity was determined in vitro. After electroporation with an optimized protocol, muRas-LCL expressed mutated ras peptides for a considerable period of time (at least 8 weeks) on the cell surface. The transfection procedure did not affect the expression of the costimulatory molecules B7.1, intercellular adhesion molecule-1, and leukocyte function associated Ag-3 by SP-LCL. muRas-LCL were able to induce muRas-specific cytotoxic T lymphocytes from three of three healthy donors and one of one patient with pancreatic carcinoma. Our results suggest that the gene transfer of muRas sequences to SP-LCL leads to an endogenous processing of this Ag in the major histocompatibility complex class I pathway and to functional presentation of antigenic peptides to CD8+ T lymphocytes. Autologous SP-LCL can serve as an unlimited renewable source for autologous cellular vaccines and appear to be good candidates as presenters for a wide range of human tumor Ags. PMID- 11023196 TI - Targeted killing of carcinoembryonic antigen (CEA)-producing cholangiocarcinoma cells by polyamidoamine dendrimer-mediated transfer of an Epstein-Barr virus (EBV)-based plasmid vector carrying the CEA promoter. AB - The present study reports a novel nonviral method to efficiently and specifically target carcinoembryonic antigen (CEA)-producing cholangiocarcinoma (CC) cells in vitro. Epstein-Barr virus (EBV)-based and conventional plasmid vectors were constructed that possess the beta-galactosidase (beta-gal) or herpes simplex virus-1 (HSV-1) thymidine kinase (Tk) genes as well as tandem repeats of the human genomic sequence -82 to -42 bp from the transcriptional start site of the CEA gene. The plasmids were transfected by means of polyamidoamine dendrimer into CEA-positive (HuCC-T1) or -negative cell lines. Transfection of the conventional plasmid vector with the CEA promoter and beta-gal gene resulted in a very low or undetectable level of marker gene expression even in the CEA-positive cell line. Transferring the HSV-1 Tk gene by conventional plasmid did not affect the susceptibility of HuCC-T1 cells to ganciclovir. In marked contrast, strong beta gal expression was specifically obtained in HuCC-T1 cells by transfecting the EBV based plasmid in which the CEA promoter and a ubiquitous promoter (SRalpha) are employed to drive the EBV-encoded nuclear antigen 1 (EBNA1) and beta-gal genes, respectively (pTES.beta). Furthermore, CEA-positive but not -negative tumor cells were rendered highly susceptible to ganciclovir when transfected with the EBV based vector that carries the CEA promoter-EBNA1 and SRalpha-HSV-1 Tk genes (pTES.Tk). These results strongly suggest that the EBV-based plasmid vector/cationic polymer system (EBV/polyplex) equipped with the CEA promoter provides an efficient nonviral method for the targeted gene therapy of CEA producing malignancies. PMID- 11023197 TI - Antineoplastic effect of anti-erbB-2 intrabody is not correlated with scFv affinity for its target. AB - Intracellular single-chain antibodies (scFvs) have emerged as a powerful method to knock out expression of oncoproteins. We have demonstrated previously that scFvs directed against a variety of molecular targets induce specific toxicity in tumor cells. Recently, the utility of an anti-erbB-2 scFv has predicated its evaluation in a phase I gene therapy clinical trial. The utility of scFv as an intrabody is closely linked to its interaction with a target, limiting the contribution of the latter to the neoplastic phenotype. In this study, we sought to determine whether improvement in the affinity of the scFv for its cognate target could improve the efficiency of intrabody-mediated oncoprotein knockout. We compared in erbB-2-positive and -negative tumor cells the function of plasmids encoding a newly developed C6.5 anti-erbB-2 scFv, which has a 1000-fold higher affinity, with our original e23 anti-erbB-2 scFv. Intracellular scFv expression, target binding, and tumor cell cytotoxicity were found to be similar in all conditions tested, including dose-response studies with limiting dilutions of the scFv. On this basis, we have concluded that the antineoplastic effect of anti erbB-2 intrabody is not correlated with scFv affinity for its target. PMID- 11023198 TI - A cell surface-displayed anti-c-myc single-chain antibody: new perspectives for the genetic improvement of cellular tumor vaccines. AB - We have shown recently that cell surface-bound, single-chain Fv antibodies (scFv) are a powerful tool for the improvement of cellular tumor vaccines. To simplify this approach and to develop a general tool for the generation and improvement of cellular tumor vaccines, we chose an scFv against a peptide from the human proto oncogene c-myc that could anchor any c-myc-tagged protein to the cell surface. The retroviral vector p50-Mx-neo (pMESV) was used to express scFv on the surface of the human melanoma line SkMel63. The cell-bound anti-c-myc scFv bound specifically to a soluble purified anti-CD28 scFv carrying a c-myc peptide-tag at its C terminus. Proof of principle was determined by incubating human peripheral blood lymphocytes with a mixture of (a) anti-c-myc-transfected SkMel63 cells binding the anti-CD28 scFv and (b) SkMel63 cells transfected with an anti-CD3 scFv. A clear synergistic effect on T-cell activation was observed that was comparable with that obtained in previous studies using SkMel63 cells transfected with the gene for the anti-CD28 scFv. As the cell surface-displayed anti-c-myc scFv can bind any c-myc-tagged protein of interest, this technique facilitates the genetic engineering of cellular vaccines for the therapy of virtually all human neoplasias. PMID- 11023199 TI - Immunological responsiveness to interleukin-2-producing brain tumors can be restored by concurrent subcutaneous transplantation of the same tumors. AB - The central nervous system shows tolerance for activated host immune reactions, and this relative unresponsiveness may lessen the efficacy of an immunotherapy for brain tumors. Using interleukin-2 (IL-2)-producing 9L rat gliosarcoma cells (9L/IL-2), we examined whether secretion of IL-2 from subcutaneous (s.c.) and/or intracerebral (i.c.) tumors can elicit augmented immunological responses to brain tumors. Syngeneic rats could reject 9L/IL-2 cells inoculated s.c., but developed 9L/IL-2 brain tumors by i.c. inoculation. The growth of i.c. 9L/IL-2 tumors was, however, significantly retarded compared with that of i.c. wild-type tumors. The growth of i.c. wild-type tumors was significantly suppressed when the rats concurrently received 9L/IL-2 cells s.c. Moreover, most of the rats that were inoculated i.c. with 9L/IL-2 cells did not develop brain tumors when concurrently injected s.c. with 9L/IL-2 cells. Immunohistochemical analysis on i.c. 9L/IL-2 tumors, when the rats were concurrently inoculated s.c. with 9L/IL-2 cells, revealed that migration of CD4+ or CD8+ T cells, monocytes/microglias, and macrophages was markedly augmented to a similar level as found in the s.c. 9L/IL 2 tumors. These results showed that systemic immune responses to brain tumor were induced in an immunologically privileged site by concurrent s.c. inoculation of the same tumors that produce IL-2. The present study may also raise the possibility of a therapeutic strategy for brain tumors by the combinatory expression of IL-2 gene using s.c. immunization followed by direct gene transfer into brain tumors. PMID- 11023200 TI - Adenovirus-mediated p16INK4 gene transfer significantly suppresses human breast cancer growth. AB - The p16INK4 tumor suppressor gene encodes a protein that inhibits cyclin dependent kinase 4, and its homologous deletion is common in human breast cancer. p16INK4 gene transfer has been reported to be efficacious in inducing growth inhibition of various human tumors such as brain, lung, prostate, and esophageal cancers. However, the efficiency of the p16INK4 gene with regard to growth inhibition of human breast cancer has not been studied extensively. To examine its tumor-suppressive function and its potential in breast cancer gene therapy, the wild-type p16INK4 gene was expressed in an adenovirus-mediated gene delivery system and introduced into breast cancer cell lines that do not express p16INK4 protein. Expression of the introduced p16INK4 blocked tumor cell entry into the S phase of the cell cycle, induced tumor cell apoptosis, and inhibited tumor cell proliferation both in vitro and in vivo. These results strongly suggest that p16INK4 is a tumor suppressor gene and suggest that it has potential utility in breast cancer gene therapy. PMID- 11023201 TI - Combination gene therapy using multiple immunomodulatory genes transferred by a defective infectious single-cycle herpes virus in squamous cell cancer. AB - Herpes simplex type 2-defective infectious single-cycle (DISC) viruses are attenuated viruses that were originally produced as viral vaccines; however, these viruses are also efficient gene transfer vehicles. The main goals of this study were to examine determinants of the gene transfer by using DISC virus for squamous cancer and to evaluate the antitumoral efficacy of vaccination with tumor cells modified by DISC viruses carrying a combination of immunomodulatory genes (interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM CSF), B7-1) in a model of squamous cell cancer (SCCVII) in C3H/HeJ mice. SCCVII cells transduced by DISC viruses (multiplicity of infection of 10) carrying the IL-2 or GM-CSF gene produced nanogram quantities of IL-2 or GM-CSF per 10(6) cells. Irradiated (5,000 cGy, 10,000 cGy) cells secreted levels of GM-CSF or IL-2 that were comparable with nonirradiated cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-IL2 or DISC-GMCSF resulted in protection against subsequent tumor challenge (P < .01), with DISC-GMCSF-transduced, irradiated tumor cells showing the greatest effects (P < .001). Marked growth arrest also was noted in established tumors after direct injection of DISC-GMCSF (P < .001). These data demonstrate that (a) DISC virus is capable of efficient gene transfer, (b) GM-CSF-secreting genetically modified tumor vaccine protects against tumor cell challenge and suppresses tumor growth, and (c) intratumoral injection of DISC-GMCSF significantly suppresses the growth of established tumors. These results not only confirm clinically relevant gene transfer but also demonstrate that the gene transfer is an effective anti-cancer therapy. PMID- 11023202 TI - Gene transfer to hepatocellular carcinoma: transduction efficacy and transgene expression kinetics by using retroviral and lentiviral vectors. AB - Gene therapy is an attractive therapy for hepatocarcinoma, and several approaches have been studied using murine leukemia virus-derived retroviruses. We compared gene transfer efficacy and transgene expression kinetics after transduction of hepatocarcinoma cell lines using enhanced green fluorescent protein (EGFP) expressing murine leukemia virus-derived retroviral vectors and HIV-derived lentiviral vectors. First, we showed that both retroviral and lentiviral vectors efficiently transduce cycling hepatocarcinoma cell lines in vitro. However, after cell cycle arrest, transduction efficacy remained the same for lentiviral vectors but it decreased by 80% for retroviral vectors. Second, we studied EGFP expression kinetics using lentiviral vectors expressing EGFP under the control of cytomegalovirus (CMV) or phosphoglycerolkinase (PGK) promoter. We show that the CMV promoter allows a stronger EGFP expression than the PGK promoter. However, in contrast to PGK-driven EGFP expression, which persists up to 2 months after transduction, CMV-driven EGFP expression rapidly decreased with time. This phenomenon is due to promoter silencing, and EGFP expression can be restored in transduced cells by using transcription activators such as interleukin-6 or phorbol myristate acetate/ionomycin and, to a lesser extent, the demethylating agent 5'-azacytidine. Altogether, our results suggest that lentiviral vectors, which allow efficient transduction of hepatocarcinoma cell lines with a strong and a sustained expression according to the promoter used, are promising tools for gene therapy of hepatocarcinomas. PMID- 11023203 TI - New practice guidelines for obstetric anesthesia. PMID- 11023204 TI - Management of stage I cervical cancer in pregnancy. AB - Although cervical carcinoma is the most common gynecologic malignancy associated with pregnancy, its occurrence is rare with an incidence of approximately 1 per 1,200 to 10,000 pregnancies. There are inadequate data addressing both the obstetric implications of the diagnostic evaluation and the impact of intervention on maternal and infant outcomes. Certain conclusions and recommendations, however, can be drawn from the available data. Diagnostic evaluation includes cytological screening, colposcopy and if necessary, biopsy, and selective conization. Staging of the pregnant patient is modified to minimize radiation exposure to the developing fetus. The treatment schema for patients with stage I cervical cancer in pregnancy varies with the stage of disease and gestational age at diagnosis. With close surveillance, deliberate delay of therapy to achieve fetal maturity is a reasonable option for patients with microinvasive and early stage IB cervical cancer. Tumor characteristics and maternal survival are not adversely affected by pregnancy. Conversely, cervical cancer does not seem adversely to affect pregnancy. However, timing and type of therapy may have a significant influence on the fate of the fetus. In counseling patients with cervical cancer during pregnancy, many factors must be considered, including the patient's desire for the pregnancy, stage of disease, and gestational age at diagnosis. Pregnant patients with stage I cervical cancer should be fully informed of all possible treatment options and consequences. The care of these patients should be closely coordinated by experts in perinatology and gynecologic oncology. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe the signs and symptoms of cervical cancer during pregnancy and the indications for conization during pregnancy, as well as being able to outline management strategies for the various stages of cervical cancer during pregnancy. PMID- 11023205 TI - Mullerian agenesis: etiology, diagnosis, and management. AB - Mullerian agenesis, a congenital malformation of the genital tract is the second most common cause of primary amenorrhea. Its etiology is poorly understood but it may be associated with renal, skeletal, and other abnormalities. The diagnosis is often made either radiologically or laparoscopically in patients in whom hormonal and karyotypic investigations for primary amenorrhea are normal. Two-dimensional ultrasound is not a reliable method of diagnosis, as exemplified by the two cases presented in this review; however, three-dimensional ultrasound may be a more sensitive diagnostic tool. The management is varied, but we conclude that the treatment of choice should be a nonsurgical approach aimed at creating a neovagina. Because of the implications for reproduction, these patients require psychological support, which should be offered as part of therapy. TARGET AUDIENCE: Gynecologists and Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe the pathophysiology and clinical presentation of mulerian agenesis and to list the other abnormalities associated with this condition and to outline potential treatment options for a patient with mullerian agenesis. PMID- 11023206 TI - Mechanism of parturition and preterm labor. AB - Preterm birth occurs in 5 to 10 percent of all pregnancies and accounts for 75 percent of neonatal mortality and morbidity, including long-term handicap. Thirty percent of preterm birth may be associated with an underlying infective process, and approximately 50 percent are idiopathic. Preterm birth corresponds to a syndrome with a variety of causes. The factors contributing to preterm birth may vary at different "windows" of gestation. Glucocorticoids seem to have a central role in processes of birth at term and preterm, and alter synthesis and/or activity of key enzymes in prostaglandin synthesis and metabolism as well as increasing corticotropin-releasing hormone output by intrauterine tissues. This new information should facilitate development of improved methods of diagnosis and therapy for patients at risk of preterm labor. TARGET AUDIENCE: Gynecologists and Family Physicians LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe potential mechanisms of partition and preterm labor and to list the various substances that can inhibit and promote myometrial contractions. PMID- 11023207 TI - A review of the potential and versatility of colloidal gold cytochemical labeling for molecular morphology. AB - In the present article we review several postembedding cytochemical techniques using the colloidal gold marker. Owing to the high atomic number of gold, the colloidal gold particles are electron dense. They are spherical in shape and can be prepared in sizes from 1 to 25 nm, which renders this marker among the best for electron microscopy. In addition, because it can be bound to several molecules, this marker has the advantage of being extremely versatile. Combined to immunoglobulins or immunoglobulin-binding proteins (protein A), it has been applied successfully in immunocytochemistry. Colloidal gold particles 5-15 nm in size are excellent for postembedding cytochemistry. Particles of smaller size, such as 1 nm, must be silver enhanced to be visualized by transmission electron microscopy. We have elected to review the superiority of indirect immunocytochemical approaches using IgG-gold or protein A-gold (protein G-gold and protein AG-gold). Lectins or enzymes can be tagged with colloidal gold particles, and the corresponding lectin-gold and enzyme-gold techniques have specific advantages and great potential. Using an indirect digoxigenin-tagged nucleotide and an antidigoxigenin probe, colloidal gold technology can also be used for in situ hybridization at the electron microscope level. Affinity characteristics lie behind all cytochemical techniques and several molecules displaying high affinity properties can also be beneficial for colloidal gold electron microscopy cytochemistry. All of these techniques can be combined in various ways to produce multiple labelings of several binding sites on the same tissue section. Colloidal gold is particulate and can easily be counted; thus the cytochemical signal can be evaluated quantitatively, introducing further advantages to the use of the colloidal gold marker. Finally, several combinations and multiple step procedures have been designed to amplify the final signal which renders the techniques more sensitive. The approaches reviewed here have been applied successfully in different fields of cell and molecular biology, cell pathology, plant biology and pathology, microbiology and virology. The potential of the approaches is emphasized in addition to different ways to assess specificity, sensitivity and accuracy of results. PMID- 11023208 TI - Melatonin circadian rhythm in the retina of mammals. AB - Melatonin has been traditionally considered to be derived principally from the pineal gland. However, several investigations have now demonstrated that melatonin synthesis occurs also in the retina (and in other organs as well) of several vertebrate classes, including mammals. As in the pineal, melatonin synthesis in the retina is elevated at night and reduced during the day. Since melatonin receptors are present in the retina and retinal melatonin does not contribute to the circulating levels, retinal melatonin probably acts locally as a neuromodulator. Melatonin synthesis in the retinas of mammals is under control of a circadian oscillator located within the retina itself, and circadian rhythms in melatonin synthesis and/or release have been described for several species of rodents. These rhythms are present in vivo, persist in vitro, are entrained by light, and are temperature compensated. The recent cloning of the gene responsible for the synthesis of the enzyme arylalkylamine N-acetyltransferase (the only enzyme unique to the melatonin synthetic pathway) will facilitate localizing the cellular site of melatonin synthesis in the retina and investigating the molecular mechanism responsible for the generation of retinal melatonin rhythmicity. Melatonin has been implicated in many retinal functions, and the levels of melatonin and dopamine appear to regulate several aspects of retinal physiology that relate to light and dark adaptation. In conclusion, it seems that retinal melatonin is involved in several functions, but its precise role is yet to be understood. PMID- 11023209 TI - Photic entrainment of circadian activity patterns in the tropical labrid fish Halichoeres chrysus. AB - Yellow wrasses (Halichoeres chrysus) show clear daily activity patterns. The fish hide in the substrate at (subjective) night, during the distinct rest phase. Initial entrainment in a 12h:12h light-dark (12:12 LD) cycle (mean period 24.02h, SD 0.27h, n = 16) was followed by a free run (mean period 24.42h, SD 1.33h) after transition into constant dim light conditions. Light pulses of a comparable intensity as used in the light part of the LD cycles did not result in significant phase shifts of the free-running rhythm in constant darkness. Application of much brighter 3h light pulses resulted in a phase-response curve (PRC) for a fish species, with pronounced phase advances during late subjective night. The PRCs differed from those mainly obtained in other vertebrate taxa by the absence of significant phase delays in the early subjective night. At that circadian phase, significant tonic effects of the light pulses caused a shortening of the circadian period length. Entrainment to skeleton photoperiods of 1:11 LD was observed in five of six wrasses exposed, also after a 3h phase advance of this LD cycle. Subsequently, a 1:11.25 LD cycle resulted in entrainment in four of the six fish. It is suggested that the expression of the circadian system in fish can be interpreted as a functional response to a weak natural zeitgeber, as present in the marine environment. This response allows photic entrainment as described here in the yellow wrasse. PMID- 11023210 TI - Age affects hibernation in Syrian hamsters (Mesocricetus auratus). AB - In this study, we aimed to show how age affects hibernation in the Syrian hamster. Experimentally, we used 30 male animals differing in age. The old animals were 20 months of age and the adults were 8 months of age at the end of the test. The young animals were 3 weeks old at the start of testing and 5 months old at the end of the testing period. The torpor observation started October 15, 1996, and ended March 11, 1997, in the laboratory colony maintained under natural photoperiod and outdoor air. Observations were performed around noon daily. Three measures (i.e., prehibernation period [hibernation latency], proportion of hibernation spent in torpor, and proportion of animals in torpor), all of which reflect the strength of occurrence of hibernation, indicated that the older hamsters (1) started hibernation earlier, (2) spent more time in torpor, and (3) had a higher chance of being in torpor than the younger ones during the hibernation season. PMID- 11023211 TI - Effect of chronic hyperprolactinemia on daily changes of glutamate and aspartate concentrations in the median eminence and different hypothalamic areas of male rats. AB - The 24h changes of glutamate (GLU) and aspartate (ASP) were studied in the median eminence (ME) and hypothalamic areas. It was analyzed whether prolactin may change their daily patterns. The hypothalamic concentration of these amino acids was measured by high-performance liquid chromatography (HPLC) with fluorometric detection. Plasma prolactin levels increased over the 24h light-dark cycle after pituitary grafting compared to controls, and its circadian rhythm was disrupted. In controls, aspartate and glutamate in the hypothalamic areas studied followed a specific daily variation or showed no rhythmicity. In the median eminence, hyperprolactinemia seem to phase advance the aspartate or glutamate peaks from 16:00 to 12:00. In the mediobasal hypothalamus, hyperprolactinemia altered daily changes of aspartate and significantly decreased its concentration. Also, it seems to delay the nocturnal glutamate peak compared to controls. In the posterior hypothalamus, hyperprolactinemia did not change aspartate and glutamate concentrations and their daily changes, although it increased the glutamine concentration. These data show the existence of 24h changes of amino acid concentration in three of the hypothalamic regions studied. Increased plasma prolactin levels differentially affected these patterns depending on the hypothalamic area analyzed. PMID- 11023212 TI - Circadian rhythms in the renin-angiotensin system and adrenal steroids may contribute to the inverse blood pressure rhythm in hypertensive TGR(mREN-2)27 rats. AB - The transgenic TGR(mREN-2)27 rat is not only characterized by fulminant hypertension, but also by a disturbance in circadian blood pressure regulation, resulting in inverse circadian blood pressure profiles. The reasons for these alterations are not very well understood at present. We therefore investigated the circadian rhythms in several hormones participating in blood pressure regulation. From TGR and Sprague-Dawley (SPRD) control rats synchronized to 12h light and 12h dark (LD 12:12) blood was collected at different circadian times (07, 11, 15, 19, 23, 03, and 07 again, 5 rats per strain and time). The activities of plasma renin and converting enzyme, as well as plasma concentrations of corticosterone and aldosterone, were determined by radioimmunoassay (RIA). SPRD rats showed significant circadian rhythms in all variables except plasma renin activity, with maxima occurring during the day. TGR rats showed significant circadian rhythmicity in plasma renin activity and corticosterone and daily variation in aldosterone; angiotensin-converting enzyme (ACE) activity did not reach statistical significance. In TGR rats, 24h means in plasma renin activity and aldosterone were approximately sevenfold and fourfold higher, respectively, than in SPRD rats. Peak concentrations in corticosterone around 15h were more than two times higher in TGR rats than in SPRD rats, whereas no differences were observed during the night. It is concluded that, in TGR rats, the overall increase in plasma renin activity and aldosterone may contribute to the elevated blood pressure. The comparatively high levels in corticosterone and plasma renin activity during daytime may be involved in the inverse circadian blood pressure profiles in the transgenic animals. PMID- 11023213 TI - Phase advance after one or three simulated dawns in humans. AB - A specially designed apparatus that can simulate the waveform of the dawn or dusk signal at any latitude and any day of the year has been shown to phase shift the circadian pacemaker in rodents and primates at a fraction of the illuminance previously used. Until recently, it was considered that rather high illuminances or rather long exposure episodes to room light were necessary to phase shift human circadian rhythms. This experiment shows that, under controlled conditions of a modified constant routine protocol, a single dawn signal is sufficient to phase advance the timing of the onset of secretion of the pineal hormone melatonin. The significant phase advance of salivary melatonin of 20 minutes, which is enhanced to 34 minutes after three consecutive dawn signals, is small, but appears to be of sufficient magnitude to entrain the human circadian pacemaker, which has an endogenous period of about 24.2h. PMID- 11023214 TI - Diurnal variation in the P300 component of human cognitive event-related potential. AB - Diurnal variation in the P300 component of the human cognitive event-related potential (ERP) was examined. The P300 component is considered to be a measure of neuroelectric activity related to cognitive functions such as attention allocation and information processing. Nine diurnally active healthy male subjects whose sleep-wake rhythms were synchronized prior to the experiment were studied. The P300 components oral temperature, heart rate, left- and right-hand grip strength, reaction time, subjectively rated sleepiness, attention level, and fatigue were measured at 08:00, 11:00, 14:00, 17:00, and 20:00. Significant diurnal variations in P300 latency, P300 amplitude, oral temperature, heart rate, left- and right-hand grip strength, subjectively rated sleepiness, and attention level were observed. The P300 latency at 08:00 was significantly longer than at 11:00, 17:00, and 20:00, while the P300 amplitude at 08:00 was significantly greater than at 17:00 and 20:00. The P300 latency was correlated positively with subjectively rated sleepiness and negatively correlated with subjectively rated attention level. These results suggest the existence of diurnal variation in human cognitive functions. PMID- 11023215 TI - Some factors influencing the sensitivity of body temperature to activity in neonates. AB - In adult humans, core temperature is influenced by activity; the sensitivity of core temperature to such effects shows a phase dependence and is also influenced by the environment and whether the individual is asleep or awake. We have investigated if similar effects are evident in neonates, in whom thermoregulation and the circadian rhythm of core temperature are not fully developed. Eleven full term, healthy babies were studied singly (light 07:00-19:00) at 2 days of age and again 4 weeks after birth; between these times, they were tended routinely on a communal ward. On study days, 10-minute recordings were made of rectal and skin (abdominal) temperature, heart rate (HR), and behavioral state. Sensitivities of the temperatures to activity ("arousal") were assessed throughout the 24h by measuring the gradient of (temperature/HR). Sensitivities measured at 01:00, 05:00, 09:00, 13:00, 17:00, and 21:00 were used as dependent variables in stepwise regression and linear regression analyses, with "subjects," "light versus dark," "behavioral state," and "difference between time of measurement and the acrophase of the endogenous component of the temperature rhythm" (ignoring sign) as possible predictors. (Acrophases of the temperature rhythms had been estimated from 24h data purified using the behavioral state record.) Light versus dark acted as a significant predictor of the sensitivity of rectal temperature to arousal on day 2 and week 4, the sensitivity increasing in the light, and there was limited evidence for behavioral state acting as a predictor on day 2. Neither factor was a significant predictor when the sensitivity of the babies' skin temperatures to arousal was investigated. There was also some evidence that the difference between the time of measurement and the temperature acrophase acted as a predictor of sensitivity to arousal in both rectal (day 2) and skin (week 4) temperature, with larger differences decreasing the sensitivity. These results indicate that there are masking effects on body temperature due to arousal in neonates, the size of which depends on both internal and external factors. However, this sensitivity of temperature to arousal shows differences from the sensitivity of temperature to physical activity in both adult humans and adult mice. One possible explanation of this result is that temperature regulation and the circadian system are not fully developed in humans at this age. PMID- 11023216 TI - Circadian rhythms in human muscular efficiency: continuous physical exercise versus continuous rest. A crossover study. AB - This study deals with the influence of time of day on neuromuscular efficiency in competitive cyclists during continuous exercise versus continuous rest. Knee extension torque was measured in ultradistance cyclists over a 24h period (13:00 to 13:00 the next day) in the laboratory. The subjects were requested to maintain a constant speed (set at 70% of their maximal aerobic speed obtained during a preliminary test) on their own bicycles, which were equipped with cyclosimulators. Every 4h, torque developed and myoelectric activity were estimated during maximal isometric voluntary contractions of knee extensors using an isokinetic dynamometer. Mesenteric temperature was monitored by telemetry. The same measures were also recorded while the subjects were resting awake until 13:00 the next day. During activity, torque changed within the 24h period (p < .005), with an acrophase at 19:10 and an amplitude of 7.8% around the mean of 70.7%. At rest, a circadian rhythm was observed in knee extensor torque (p < .05), with an acrophase at 19:30 and an amplitude of 6% around the mean of 92.3%. Despite the standardized conditions, the results showed that isometric maximal strength varied with time of day during both a submaximal exercise and at rest without prior exercise. The sine waves representing these two rhythms were correlated significantly. Although at rest the diurnal rhythm followed muscular activity (i.e., neurophysiological factors), during exercise, this rhythm was thought to stem more from fluctuations in the contractile state of muscle. PMID- 11023217 TI - Circadian variation in onset of acute cardiogenic pulmonary edema is independent of patients' features and underlying pathophysiological causes. AB - BACKGROUND: The present study aimed to confirm the existence of a circadian pattern in the onset of acute pulmonary edema (APE) and to verify whether sex, age, preexisting diseases, and clinical causes determining the event may influence it. SUBJECTS AND METHODS: The study considered all consecutive cases of APE observed at the St. Anna General Hospital of Ferrara, Italy, during a 7-year period from January 1, 1992, to December 31, 1998. The sample population was divided into subgroups by sex, age (<75 and > or =75 years), presence or absence of diabetes and hypertension, clinical causes determining the event (i.e., acute myocardial infarction (AMI), pulmonary embolism, arrhythmias). The most important associated or concomitant diseases were also considered (i.e., coronary heart disease and angina, previous myocardial infarction, chronic cardiac failure, dilatative cardiopathy, chronic atrial fibrillation, valvular disease, chronic obstructive pulmonary disease, chronic cor pulmonale, malignancy, chronic renal failure). Time of symptom onset of each event was recorded accurately, then tabulated into 24 increments of 1h (e.g., 06:00 to 06:59 was reported as 6 A.M.). For statistical chronobiological analysis, partial Fourier series were used. RESULTS: During the 7-year period, 1321 consecutive cases of APE in 1014 different subjects were observed. The majority of events occurred at night, and statistical analysis showed a 24h rhythmicity both in the total sample population and in all considered subgroups, with the only exception being patients with pulmonary embolism and arrhythmias, for which the small number of cases made the study of rhythms in APE impossible. CONCLUSIONS: The nighttime preference in the occurrence of APE appears to be quite independent of all demographic features or underlying pathophysiological causes. PMID- 11023218 TI - Subtalar arthrodesis with flexor digitorum longus transfer and spring ligament repair for treatment of posterior tibial tendon insufficiency. AB - The surgical treatment of flexible pes planovalgus deformities resulting from Stage 2 posterior tibial tendon insufficiency is controversial and many techniques have been proposed. We retrospectively reviewed the results of subtalar arthrodesis combined with spring ligament repair/reefing and flexor digitorum longus (FDL) transfer to the navicular. There were sixteen patients (seventeen feet) with an average follow-up of 27 months (9-52). All deformities were passively correctable. The average age was 56 yrs (39-78). All patients had failed conservative management, 88% had previously been treated with orthotics, and 53% had lateral pain from subfibular impingement. Two patients were noted to have degenerative changes of the subtalar joint. Successful subtalar joint fusion occurred in all patients with an average time to radiographic union of 10.1 weeks (5-24). The average AOFAS hindfoot score and Maryland foot score postoperatively was 82 and 86 respectively. Standing radiographic analysis demonstrated an average improvement in the AP talo-1st metatarsal angle of 6 degrees (24 degrees preoperative, 18 degrees postoperative). The talonavicular coverage angle improved an average of 17 degrees (34 degrees preoperative, 17 degrees postoperative). The lateral talo-1st metatarsal angle improved an average of 10 degrees (18 degrees preoperative, 8 degrees postoperative). The lateral talocalcaneal angle decreased an average of 21o (55 degrees preoperative, 34 degrees postoperative). The distance of the medial cuneiform to the floor on the lateral radiograph averaged 12mm preoperatively and 18mm postoperatively (avg. improvement 6mm). The combination of the flexor digitorum longus tendon transfer and spring ligament repair with subtalar arthrodesis is an effective and reliable procedure which provides excellent correction of hindfoot valgus as well as forefoot abduction and restoration of the height of the longitudinal arch. These results compare favorably with flexor transfer combined with either calcaneal osteotomy or lateral column lengthening. PMID- 11023219 TI - Calcaneocuboid joint pressure after lateral column lengthening in a cadaveric planovalgus deformity model. AB - The purpose of this study is twofold: first, to measure the joint contact pressure across the calcaneocuboid joint in a planovalgus deformity and compare the results to pressures measured in a normal foot; and second, to determine the change in pressure across the calcaneocuboid joint after an Evan's-type calcaneal lengthening osteotomy. The effect of this procedure on the calcaneocuboid joint was evaluated using seven cadaver feet to measure peak pressure across the calcaneocuboid joint under a constant load. Each foot was sectioned medially to reproduce a deformity consistent with an adult, acquired flatfoot. Each flatfoot deformity was then corrected using a ten-millimeter lateral column lengthening osteotomy. Joint pressures were measured in the normal foot, the created flatfoot and then in the corrected flatfoot. Peak pressures across the joint increased significantly from baseline in the flatfoot (p <0.05). However, the change in pressure from the flatfoot to the corrected foot was not significant, and in some cases peak pressures in the corrected foot were actually lower than in the flatfoot. These findings indicate that calcaneal lengthening through an Evan's osteotomy does not increase pressure across the calcaneocuboid joint beyond physiologic loads in the flatfoot. PMID- 11023220 TI - Biomechanical comparison of syndesmosis fixation with 3.5- and 4.5-millimeter stainless steel screws. AB - Although most authors recommend either 3.5-mm or 4.5-mm cortical screws for syndesmosis fixation, the optimum screw size has yet to be defined. The present study was designed to biomechanically compare syndesmosis fixation with 3.5-mm and 4.5-mm stainless steel screws. Simulated pronation external rotation ankle injuries were created in twelve paired, fresh-frozen cadaveric leg specimens. One limb from each pair received a 3.5-mm tricortical stainless steel screw for syndesmosis fixation (group I), while the contralateral specimen was stabilized using a 4.5-mm screw (group II). Sub-maximal axial ramp (0 to 1200 N) and external rotation/torsional ramp (0 to 5 N-m) loading was performed on each specimen prior to ligament division, following ligament division and following syndesmosis fixation. Axial fatigue testing was then performed at 1.5 Hz for a total of 100,000 cycles (0 to 900 N), and each specimen was subsequently tested to failure in external rotation. Ligament division resulted in syndesmosis widening (p<0.001) and reduced stiffness (p<0.001) during torsional ramp loading. Subsequent syndesmosis screw placement reduced syndesmosis widening (p<0.05) and increased stiffness (p<0.05). Following screw fixation, however, widening remained greater (p<0.005) and stiffness less (p<0.001) than pre-injury levels. No differences between groups I and II were observed during submaximal testing. In external rotation to failure testing, group I failed at a greater angle (38.9 degrees +/- 4.1 degrees vs. 32.0 degrees +/- 3.8 degrees in group II; p<0.05). Failure torque was slightly higher in group I; however, the difference was not statistically significant (17.8 +/- 2.0 N-m vs. 14.3 +/- 2.6 N-m in group II; p=0.082). Five specimens in group I failed by screw pullout and five specimens in group II failed by fibula fracture (p=0.061). The present results suggest that there is no biomechanical advantage of a 4.5-mm screw over a 3.5-mm in fixation of the syndesmosis. PMID- 11023221 TI - Limited intertarsal arthrodesis using bone grafting and pin fixation. AB - Iliac crest bone grafting and K wire fixation was used to fuse the arthritic intertarsal articulations of sixteen patients with post-traumatic or primary degenerative disease. Using pre- and post-operative AOFAS scores, foot function indices (FFI), and a subjective scoring system, functional and subjective outcome was critically evaluated. The AOFAS score improved an average of 41 points, the FFI decreased from 64 to 24, and 81 percent of patients rated their results as good or excellent. Fusion of the 4th and 5th TMT joints was associated with significantly poorer functional scores, and people with worker's compensation claims had poorer subjective scores. The authors demonstrated comparable results to previously published series using other methods of fixation. PMID- 11023222 TI - Reliability and running speed effects of in-shoe loading measurements during slow treadmill running. AB - In-shoe measurement systems allow the clinician and researcher to examine the loading parameters within the shoe. This study sought to investigate the test retest reliability and speed effects of in-shoe loading parameters using the Pedar System (Novel GMBH Munich) during slow treadmill running. The results indicated good to excellent test retest reliability between the two days test Intraclass correlation coefficients (ICC's) ranged from 0.84-0.99 depending on the plantar region and variable analyzed. All plantar loading variables increased (peak pressure, peak pressure time impulse, peak force, and force time impulse) with the exception of contact area when treadmill running speed was increased from 2.24 m/s to 3.13 m/s. Results indicate that control of running speed is essential in obtaining reproducible data using this system to measure in-shoe loading data. PMID- 11023223 TI - The ANK device: a new approach in the treatment of the fractures of the lateral malleolus associated with the rupture of the syndesmosis. AB - The ANK device was developed for the treatment of fractures of the lateral malleolus occurring with rupture of the syndesmosis. While it provides the anatomic reduction of the fracture and the syndesmosis, it allows the physiologic movements of the fibula. It is not used for comminuted fractures of the lateral malleolus and in cases where fibular medullary canal is narrow. We included forty nine patients who had the ANK device applied and at least 2 years follow-up. The mean follow-up was 41 months (range 24-124). The fractures were evaluated according to the Lauge-Hansen classification; 25 cases were evaluated as supination-external rotation, 11 cases were pronation-abduction, and 13 cases were pronation-external rotation type fractures. There were also 46 fractures of the medial malleolus and three ruptures of the deltoid ligament. Twenty-nine (59,2%) patients were evaluated as excellent, 12 (24,5%) as good, 5 (10.2%) as fair and 3 (6.1%) as poor. Arthrosis was observed in 3 (6.1%) of the patients. PMID- 11023224 TI - Influence of foot, leg and shoe characteristics on subjective comfort. AB - The purpose of this study was to determine the relationships between foot and leg characteristics, shoe characteristics, and the short-term subjective comfort of three different pairs of athletic shoes. Static measurements of foot dimension and leg angles were taken from eighteen subjects. Subjects rated the comfort of three different athletic shoes for standing, walking and running. The shoes were quantified by internal dimensions, hardness, flexibility and torsional stiffness. Average comfort ratings decreased from standing to walking to running. One shoe seemed suited for only a small group of subjects. In contrast, another shoe was generally comfortable for a large group. Skeletal alignment, specifically eversion angle, was related to comfort for one shoe. Therefore, fit of the shoe is not sufficient for comfort. Skeletal alignment, shoe torsional stiffness and cushioning seem to be mechanical variables which may be important for comfort. PMID- 11023225 TI - Skewfoot in patients with osteogenesis imperfecta. AB - We report skewfoot deformities in two patients who have osteogenesis imperfecta. A discussion will follow proposing etiologies of skewfoot, speculating that the ligamentous laxity often present in children who have osteogenesis imperfecta may predispose the development of skewfoot. PMID- 11023226 TI - Effects of lower extremity rotation on prognosis of flexible flatfoot in children. AB - The aim of this study is to search the effects of tibial torsion on flexible flatfoot. For this purpose, 20 children with flexible flatfoot and external tibial torsion were determined as a study group. The control group which consisted of 10 children with flexible flatfoot without rotational problems was included in the study. Plantar flexion angle of talus, talo-1. metatarsal angle and dorsoplantar talocalcaneal angle were measured on standing foot radiographs. Tibial torsion was measured by computed tomography. Plantar flexion angle of talus and dorsoplantar talocalcaneal angle were found significantly lower in the study group (p<0.005 and p<0.005 respectively), although talo-1. metatarsal angle was not significantly different between the groups (p=.2917). Naviculocuneiform sag was prominent in all cases of study group. In conclusion, abnormal external tibial torsion may affect the foot deformity and this can change the benign nature of the flexible flatfoot. PMID- 11023227 TI - Elastofibroma of the foot: uncommon presentation: a case report and review of the literature. AB - We report the clinical, microscopic and ultrastructural features of an elastofibroma arising in the foot. The lesion typically occurs in the elderly, and in 85% of cases arises from the connective tissue of the posterior chest wall. The histopathologic features of this lesion are distinctive, and are characterized by a haphazard array of eosinophilic collagen and elastic fibers, associated with fibroblasts and aggregates of mature fat cells. There are only two reported cases in the literature arising in the foot. PMID- 11023228 TI - A technique for harvesting bone graft for arthrodeses around the ankle. PMID- 11023229 TI - A technique for distal tibial bone graft for arthrodesis of the foot and ankle. AB - Hindfoot and forefoot arthrodeses procedures are commonly performed using autogenous bone graft harvested from the distal tibia. A new technique is described for obtaining cancellous bone graft from the distal tibia using a bone graft harvesting system specifically designed to obtain morsellized material through a small incision site. The use of this device in association with low dose fluoroscopy and backfilling of the defect with osteoconductive synthetic graft material is also discussed. PMID- 11023230 TI - Correlative high-resolution morphologic analysis of the three-dimensional organization of human chromosomes. AB - A correlative morphologic analysis was carried out on isolated metaphase chromosomes by means of field emission in-lens scanning electron microscopy (FEISEM) and atomic force microscopy (AFM). Whereas FEISEM provides ultra-high resolution power and allows the surface analysis of biological structures free of any conductive coating, the AFM allows imaging of biological specimens in ambient as well as in physiologic conditions. The analysis of the same samples was made possible by the use of electrical conductive and light transparent ITO glass as specimen holder. Further preparation of the specimen specific for the instrumentation was not required. Both techniques show a high correlation of the respective morphologic information, improving their reciprocal biological significance. In particular, the biological coat represents a barrier for surface morphologic analysis of chromosome spreads and it is sensitive to protease treatment. The chemical removal of this layer permits high-resolution imaging of the chromatid fibers but at the same time alters the chromosomal dimension after rehydration. The high-resolution level, necessary to obtain a precise physical mapping of the genome that the new instruments such as FEISEM and AFM could offer, requires homogeneously cleaned samples with a high grade of reproducibility. A correlative microscopical approach that utilizes completely different physical probes provides complementary useful information for the understanding of the biological, chemical, and physical characteristics of the samples and can be applied to optimize the chromosome preparations for further improvement of the knowledge about spatial genome organization. PMID- 11023231 TI - Inexpensive, high-quality optical relay for use in confocal scanning beam imaging AB - An inexpensive, high optical-quality relay lens made up of two eyepieces arranged in an afocal assembly for use in confocal scanning laser imaging is described. In the past we have used relays, within our confocal microscopes, made up of achromats with long focal lengths (> or = 10 cm), which take up large optical tracks and suffer from significant amounts of astigmatism and curvature of field. We quantify aberrations associated with achromat and eyepiece relays using CODE V optical design and analysis software. The eyepiece relay is found to be more compact, better corrected, and not significantly more expensive than its achromat counterpart. In addition to being used to interconnect two scanning mirrors optically as well as scanning mirrors with microscope objectives, it can form part of the optics in a confocal scanning laser MACROscope-Microscope system (Biomedical Photometrics, Inc., Waterloo, Ontario, Canada). Due to design constraints, the MACROscope-Microscope system cannot incorporate a conventional wide-field microscope into its structure such as is done in most commercial confocal microscopes. The eyepiece relay is used as a stand-alone, compact optical link between the scanning mirrors and the microscope objective. This consequently makes the MACROscope-Microscope system more compact and easier to commercialize. PMID- 11023232 TI - Visualization of artificial lignin supramolecular structures AB - In this paper we are presenting the results of our environmental scanning electron microscopy (ESEM) investigation of the lignin model compound- enzymatically polymerized coniferyl alcohol, also known as dehydrogenate polymer (DHP). The goals of this study were to visualize the supramolecular organization of DHP polymer on various substrates, namely graphite, mica, and glass, and to explore the influence of substrate surface properties and associated collective phenomena on the lignin self-assembled supramolecular structure. Based on results obtained with ESEM, combined with previously published results based on scanning tunneling microscopy (STM) and electron spin resonance (ESR) technique, we looked at lignin structure ranging from a monomer on a fraction of nanometer scale to a large aggregate on a fraction of millimeter scale, therefore using six orders of magnitude range of size. Herein, we are presenting evidence that there are at least four different levels of the supramolecular structure of lignin, and that its supramolecular organization is well dependent on the substrate surface characteristics, such as hydrophobicity, delocalized orbitals, and surface-free energy. PMID- 11023234 TI - Polymer-based materials to be used as the active element in microsensors: a scanning force microscopy study AB - Polymer-based materials can be incorporated as the active sensing elements in chemiresistor devices. Most of these devices take advantage of the fact that certain polymers will swell when exposed to gaseous analytes. To measure this response, a conducting material such as carbon black is incorporated within the nonconducting polymer matrix. In response to analytes, polymer swelling results in a measurable change in the conductivity of the polymer/carbon composite material. Arrays of these sensors may be used in conjunction with pattern recognition techniques for purposes of analyte recognition and quantification. We have used the technique of scanning force microscopy (SFM) to investigate microstructural changes in carbon-polymer composites formed from the polymers poly (isobutylene) (PIB), poly (vinyl alcohol) (PVA), and poly (ethylene-vinyl acetate) (PEVA) when exposed to the analytes hexane, toluene, water, ethanol, and acetone. Using phase-contrast imaging (PI), changes in the carbon nanoparticle distribution on the surface of the polymer matrix are measured as the polymers are exposed to the analytes in vapor phase. In some but not all cases, the changes were reversible (at the scale of the SFM measurements) upon removal of the analyte vapor. In this paper, we also describe a new type of microsensor based on piezoresistive microcantilever technology. With these new devices, polymeric volume changes accompanying exposure to analyte vapor are measured directly by a piezoresistive microcantilever in direct contact with the polymer. These devices may offer a number of advantages over standard chemiresistor-based sensors. PMID- 11023233 TI - Field-emission scanning electron microscopy of the internal cellular organization of fungi. AB - Internal viewing of the cellular organization of hyphae by scanning electron microscopy is an alternative to observing sectioned fungal material with a transmission electron microscope. To study cytoplasmic organelles in the hyphal cells of fungi by SEM, colonies were chemically fixed with glutaraldehyde and osmium tetroxide and then immersed in dimethyl sulfoxide. Following this procedure, the colonies were frozen and fractured on a liquid nitrogen-precooled metal block. Next, the fractured samples were macerated in diluted osmium tetroxide to remove the cytoplasmic matrix and subsequently dehydrated by freeze substitution in methanol. After critical point drying, mounting, and sputter coating, fractured cells of several basidiomycetes were imaged with field emission SEM. This procedure produced clear images of elongated and spherical mitochondria, the nucleus, intravacuolar structures, tubular- and plate-like endoplasmic reticulum, and different types of septal pore caps. This method is a powerful approach for studying the intracellular ultrastructure of fungi by SEM. PMID- 11023235 TI - Atomic force microscopy of human hair. AB - The atomic force microscope (AFM) was used to investigate the surface architecture of the entire lengths of cleaned human head hairs. Many features previously seen with the scanning electron microscope (SEM) were identified. However, the AFM has provided much greater detail and, in particular, the hair's cuticular surfaces appear not to be as smooth as had been previously supposed. A consistent feature was of step discontinuities or "ghosts" on the scale surfaces. These delineated the original location of each overlying scale before its edge had been chipped away. There was a change in the longitudinal angular presentation of the surfaces about each ghost. This means the distal ends of each cuticle cell have been synthesised in the follicle to be thicker than where that same cuticle cell is bounded on both sides by other cuticle cells. The undamaged outer cuticular surfaces at the root end of each hair were covered everywhere by longitudinal ridges (striations). Where the hair surface was worn, the striations terminated at a scale edge ghost. The ridges were approximately 9 nm high and were in parallel array with a lateral repeat spacing of about 350 nm. The striations are evidently formed on the outer surface of each cuticle cell following earlier contact in the hair follicle with the inner root sheath. The study of stained transverse sections of hairs in the transmission electron microscope (TEM) is suggested as a means for throwing some light on the underlying structure and chemistry of the striations. Finally, our AFM studies have revealed that the surface of the freshly emergent hair gradually changes over a distance of about 20 mm and that the surface of the hair for most of its length is quite different from that near the root. This is likely to be of import to those engaged in the hair toiletries industry. PMID- 11023236 TI - The effects of space charge on contrast in images obtained using the environmental scanning electron microscope AB - We present experimental evidence for the existence of a space charge in the environmental scanning electron microscope. Space charge formation is attributed to differences in the mobilities of negative and positive charge carriers in the imaging gas. A model is proposed for the behavior of space charge during image acquisition. The effects of space charge on images acquired using the gaseous secondary electron detector, ion current, and backscattered electron signals are interpreted using the proposed model. PMID- 11023237 TI - Electron microscopy of squamous cell carcinoma of the head and neck. AB - Squamous cell carcinoma of the head and neck carries a bad prognosis. In order to achieve cure, the most important thing to attain is local tumour control. The main therapy available is external radiotherapy, which can be supplemented when necessary, with interstitial radiotherapy, chemotherapy and surgery. In this paper we have evaluated specimens, taken before therapy, from 35 patients with squamous cell carcinoma of the head and neck. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses were made. With SEM, the parameters analysed were the amount and appearance of microvilli, filaments, and blood vessels. From TEM, scoring was made of the filaments, desmosomes, nuclei, nucleoli, mitochondria, and blood vessels. Scoring of the samples showed a difference between the group with recurrent disease (n = 10, Group 1) and the group with local tumor control (n = 25, Group 2) in regard to both blood vessels and intracellular filaments. No differences of the nuclei, nucleoli, or the mitochondria were observed. PMID- 11023238 TI - The potential prognostic value of ultrasonic characterization (videodensitometry) of myocardial tissue in essential arterial hypertension. AB - BACKGROUND: Left ventricular hypertrophy (LVH) and the geometric shape of the left ventricle are well-established important risk factors for cardiovascular morbidity and mortality in the hypertensive population. Videodensitometry is an alternate echocardiographic approach to the study of myocardial structural and functional alterations in essential hypertension. OBJECTIVES: To analyze the behavior of the ultrasonic videodensitometric parameter for various subgroups of a hypertensive population; first according to the severity of LVH (group A, without LVH; group B, with mild-to-moderate LVH; and group C, with severe LVH) and second according to geometric adaptation of left ventricle to pressure-volume overload of essential hypertension (group NG, normal geometry; group CR, concentric remodeling; group CH, concentric hypertrophy; and group EH, eccentric hypertrophy). METHODS: For 70 male, essential hypertensive patients and 32 normotensive healthy subjects matched for age (58 +/- 7 years) and sex as controls (group N) we performed ambulatory blood pressure measurements for the evaluation of 24 h mean systolic and diastolic blood pressures, conventional two dimensional Doppler echocardiography to evaluate left ventricular performance and left ventricular mass index, and digitization of left ventricular parasternal long-axis echocardiographic images. For regions of interest selected within the septum and the posterior wall, the mean gray levels were calculated at end systole and end-diastole. The resulting values were used to estimate the percentage cyclic variation index (CVI). RESULTS: The results according to left ventricular mass index were CVI for septum group N 34.7 + 16.3%; group A - 0.18 +/- 16%, group B - 13 +/- 19%, and group C - 22 +/- 12% (P < 0.001); and CVI of posterior wall, group N 38.2 +/- 15.4%, group A -0.75 +/- 16%, group B -16 +/- 16% and group C -16 +/- 13% (P< 0.001). According to left ventricular geometry CVI for septum were group NG 0.6 +/- 24%, group CR 1.9 +/- 17%; group CH - 25.4 +/- 18%, and group EH -17.1 +/- 20% (P < 0.01). CVI of posterior wall were group NH -5.8 + 24%, group CR 6.4 +/- 23%, group CH -29 +/- 20%, group EH -20 +/- 21 (P < 0.01). CONCLUSIONS: Our results demonstrate that subjects with high left ventricular masses and those with concentric hypertrophy, which have the worst prognostic impacts, have the most significant changes in CVI. Furthermore, videodensitometric findings are quite different even among the subgroups with mild-to-moderate left ventricular hypertrophy and eccentric hypertrophy. Therefore this videodensitometric approach could provide some useful information for better definition of cardiovascular risk in hypertension. PMID- 11023239 TI - Greater than normal prevalence of seropositivity for Helicobacter pylori among patients who have suffered myocardial infarction. AB - BACKGROUND: There is evidence to suggest that inflammation plays a role in the development of atherosclerosis. Chronic infections may activate an inflammatory response in the walls of blood vessels. OBJECTIVE: To investigate the possibility of there being an association between infection with Helicobacter pylori (H. pylori) and coronary heart disease. METHODS: We examined 100 consecutive patients documented to have recently suffered acute myocardial infarction and 100 control subjects from the same geographical area for whom there was no evidence of coronary heart disease, carefully matched both for age and sex. Blood samples were tested for the presence of immunoglobulin G antibodies against H. pylori with a serological test. RESULTS: In comparison with controls, patients were more commonly smokers (26 versus 12%/0, P < 0.05) and had more commonly been treated for hypertension (37 versus 20%, P< 0.01). There was a significant association between seropositivity for H. pylori and having previously suffered acute myocardial infarction (68 versus 53%, odds ratio 1.36 with 95% confidence interval 1.02-1.82, P=0.034). These findings remained valid in a multivariate analysis including possible confounding factors (age, sex, smoking and hypertension; odds ratio 1.35 with 95% confidence interval 1.01-1.83, P=0.046). CONCLUSIONS: The positive association between seropositivity for H. pylori and having previously suffered acute myocardial infarction found in this study provides further support for the hypothesis that there is a causal association between chronic infection with H. pylori and the development of coronary heart disease. PMID- 11023240 TI - Women react with more myocardial ischemia and angina pectoris during elective percutaneous transluminal coronary angioplasty. AB - BACKGROUND: Women have been considered to be at higher risk of complications relating to percutaneous transluminal coronary angioplasty (PTCA) than are men. One reason for this sex-related difference could be the ischemic response of myocardium during the procedure. OBJECTIVE: To investigate whether there are sex related differences in ischemic response of myocardium during elective PTCA. METHODS: Consecutive patients (n = 192, of whom 48 were women), were subjected to vectorcardiography during the PTCA procedure. Vectorcardiographic variables, magnitude of ST-segment vector (ST-VM), and magnitude of ST-segment vector change (STC-VM) were studied. RESULTS: Women were older (63 +/- 10 versus 56 +/- 10 years, P< 0.001) than men in our study and more often had diabetes mellitus and hypertension. Women less often had stents implanted (24 versus 50%, P < 0.01) and they were subjected to fewer balloon inflations (P < 0.001), with a total inflation time shorter than that for men (P< 0.001). Maximum STC-VM was 25% greater for women (P < 0.05). Women reported greater maximum pain (P < 0.05) and nitroglycerine was more frequently used for them during PTCA (P < 0.05). Occurrence of episodes of residual ischemic STC-VM (the difference between total number of episodes and number of balloon inflations) was more common for women (3 +/- 5 versus 1 +/- 3, P< 0.01). Duration of residual ischemic STC-VM episodes (the difference between total duration of episodes and duration of balloon inflations) was longer for women than it was for men (242 +/- 275 versus 148 +/- 233 s, P < 0.05). In a stepwise multivariate analysis and for a matched case control group, episodes of residual STC-VM and duration of residual STC-VM episodes still indicated that there was an independent sex-related difference (P < 0.01 and P < 0.01, respectively). CONCLUSIONS: Women more commonly develop vectorcardiographic signs of severe myocardial ischemia, more frequently experience episodes of ischemia and report more severe angina pectoris during elective PTCA than do men. PMID- 11023242 TI - Value of dobutamine stress echocardiography for diagnosis of coronary artery disease in patients with left bundle branch blockage. AB - BACKGROUND: Left bundle branch blockage (LBBB) is a strong predictor of cardiovascular mortality. Non-invasive tests such as exercise-stress testing and scintigraphy studies have no diagnostic value for diagnosis of coronary artery disease (CAD) in patients with LBBB. OBJECTIVE: To study the role of dobutamine stress echocardiography (DSE) in identification of CAD in patients with LBBB. METHODS: Thirty patients (19 men and 11 women, mean age 60 +/- 8 years) with permanent, complete LBBB were studied prospectively with DSE and coronary angiography. RESULTS: Results of DSE were compared with results of coronary angiography for left anterior descending artery and either left circumflex or right coronary artery territories, or both. Significant CAD was found in left anterior descending coronary arteries in 11 patients by coronary angiography; nine of whom were identified by DSE. Significant left circumflex or right coronary artery disease, or both, was found in nine patients; eight of whom were identified by DSE. The sensitivity, specificity and accuracy of DSE for identifying CAD in left anterior descending coronary artery territory were 82, 95 and 90%, respectively. For identifying CAD in the circumflex and right coronary artery territories sensitivity, specificity and accuracy were 88, 96 and 93%, respectively. CONCLUSION: We concluded that DSE is a very sensitive, specific and accurate non-invasive test for identification of CAD, both in left anterior descending and in left circumflex and right coronary artery territories of patients with LBBB. PMID- 11023241 TI - Activation of myocardial angiogenesis and upregulation of fibroblast growth factor-2 in transmyocardial-revascularization-treated mice. AB - OBJECTIVE: To evaluate the growth factor responses associated with myocardial angiogenesis. DESIGN: Mice were treated with transmyocardial revascularization (TMR) and evaluated for angiogenic and growth factor responses. METHODS: TMR was performed via thoractomy with a 27 g needle. At 2, 5, and 7 days post-treatment, hearts were removed from the TMR treated and control groups, then assayed for angiogenesis, fibroblast growth factor (FGF)-2 expression and vascular endothelial cell growth factor (VEGF) expression. RESULTS: TMR caused an angiogenic reaction in the myocardial blood vessels at 7 days post-TMR treatment. Elevated FGF-2 corresponded with increased TMR related angiogenesis. VEGF levels only increased in hearts that were prewounded then TMR treated. CONCLUSIONS: The data show that TMR stimulates myocardial angiogenesis. The angiogenic reaction is mediated by FGF-2 which increased in most experimental treatment groups. The VEGF response was more specific, requiring prewounding then TMR treatment for a VEGF increase. PMID- 11023243 TI - Transpulmonary contrast echocardiography: effects on delineation of endocardial border, assessment of wall motion and interobserver variability in stress echocardiograms of limited image quality. AB - BACKGROUND: A major limitation of stress echocardiography remains poor image quality. OBJECTIVE: To investigate the effects of transpulmonary contrast echocardiography (TCE) with BY 963 on endocardial border delineation, detectability of wall motion abnormalities and interobserver variability at rest and during dobutamine stress echocardiography (DSE) in subjects with technically limited baseline echocardiograms. METHODS: BY 963 was administered intravenously to 36 patients (5 ml for parasternal LAX/SAX, 10 ml for apical four-chamber/two chamber view) both at rest and at peak stress during DSE. Two observers applied a delineation score (0, endocardial border not visible; 1 border poorly visible; and 2, border clearly visible) to 12 wall segments in the parasternal and 10 in the apical views both before and after administration of BY 963. A 16-segment wall-motion score was used. RESULTS: In parasternal views, the delineation score was not improved by TCE. In the apical views, TCE significantly increased the delineation score (from 14.1 +/- 5.4 to 20.7 +/- 4.2 at rest and from 14.6 +/- 5.7 to 21.7 +/- 4.1 under stress, both P< 0.01). For 18 of 25 patients with coronary artery disease (> or = 70% stenosis) results of DSE were positive before TCE, whereas results were positive for 21 patients during TCE. For 10 of 11 patients without coronary artery disease, results of DSE were negative both before and during TCE. For the apical delineation score, interobserver variability was decreased significantly by TCE (from 19.5 +/- 19.6 to 8.2 +/- 15.6% at rest and from 20.2 +/- 19.6 to 3.3 +/- 11.4% at peak stress, both P< 0.01). CONCLUSIONS: TCE enhances endocardial border delineation in apical views at rest and during DSE, resulting in a decrease of interobserver variability and an improvement in assessment of wall motion. Use of TCE, at least how it was applied in this investigation, seems not to be indicated for parasternal projections. PMID- 11023244 TI - Reproducibility of serial intravascular ultrasound measurements in patients with angiographically silent coronary artery disease after heart transplantation. AB - BACKGROUND: Intracoronary ultrasound (ICUS) imaging is the most sensitive method for the early detection and serial evaluation of vasculopathy of transplants. Both lack of agreement between observers and lack of agreement between serial, independent pullback procedures (repeatability), which can result in a variable intraluminal catheter position may limit the reproducibility of ICUS measurements. OBJECTIVE: To evaluate the reproducibility of serial measurements of standard linear and area cross-sectional coronary dimensions in patients with non-obstructive transplant vasculopathy. METHODS: We performed ICUS imaging of patients without angiographic evidence of obstructive epicardial coronary artery disease after heart transplantation. A 30 MHz phased-array transducer was used. Two independent pullbacks of the left anterior descending coronary artery were performed and recorded on CD-ROM for off-line quantitative analysis of the most severely diseased site. Agreement of observers and repeatability of serial measurements were calculated by the use of linear regression analysis and Bland Altman plots. RESULTS: Regarding agreement of observers, correlation coefficients for intra-observer agreement ranged from r = 0.98 to r = 0.99; those for interobserver agreement ranged from r = 0.87 to r = 0.98. Serial measurements of the identical coronary artery cross-section within independent catheter pullback procedures were possible for 104 of 112 target lesions (92.90/%). Correlation coefficients ranged from r = 0.91 to r = 0.97 (for lumen diameter r = 0.91, for lumen area r = 0.93, for vessel diameter r = 0.91, for vessel area r = 0.97, for thickness of plaque r = 0.96 and for area of plaque 0.94). The mean difference of measurements was around zero for all parameters with SD from 0.13 to 0.4 mm for linear parameters and from 1.53 to 1.82 mm2 for area parameters. CONCLUSION: Serial intravascular ultrasound measurements are highly reproducible without any evidence of systematic error and a SD of differences of measurements beyond the maximal spatial resolution of currently available intravascular ultrasound catheters. PMID- 11023246 TI - Bibliography. Current world literature. PMID- 11023245 TI - Human platelet alphaIIbeta3 integrin binding affinity and specificity of SJ874: antiplatelet efficacy versus aspirin. AB - OBJECTIVE: To define the affinity and specificity of SJ874, a nonpeptide antiplatelet agent for platelet glycoprotein Ilb/IIIa integrin, and to determine the antiplatelet efficacy of SJ874 relative to those of glycoprotein IIbIIIa antagonists and aspirin. METHODS: Binding affinity and specificity of SJ874 for platelet glycoprotein IIb/IIIa integrin were determined using integrin-mediated binding and adhesion assays with human cells. Additionally, the antiplatelet efficacy of SJ874 was determined and compared with those of other glycoprotein IIb/IIIa antagonists and aspirin using light-transmittance and laser-scattering aggregometry. RESULTS: SJ874 inhibited aggregation of human platelets induced by 10 micromol/l adenosine diphosphate (ADP) with a concentration for half-maximal effect of 0.046 +/- 0.005 micromol/l using light-transmittance aggregometry. Using laser-scattering aggregometry, SJ874 was found to totally inhibit formation both of micro-aggregates and of macro-aggregates induced either by ADP or by epinephrine. In contrast, administration of 325 mg aspirin to normal healthy volunteers attenuated formation of macro-aggregates but not micro-aggregates. SJ874 inhibited binding of [125I]-fibrinogen to activated (by ADP, epinephrine, and arachidonic acid at concentrations of 100 micromol/l each) gel-filtered human platelets with a concentration for half-maximal effect of 0.0012 +/- 0.0005 micromol/l. SJ874 was demonstrated to associate more tightly with resting human platelets than did DMP754 [1] and slightly less tightly than did DMP802 [2]. SJ874 was demonstrated to exhibit a high degree of specificity for platelet glycoprotein IIb/IIIa (alphaIIb/beta3) integrin compared with other known integrins, including alphavbeta3, alphavbeta5, and alpha5beta1 (concentration for half-maximal effect > 100 micromol/l). CONCLUSION: SJ874 is a potent and specific platelet glycoprotein IIb/IIIa antagonist with high affinity for and tight association with human platelets. These data suggest that SJ874 might have good antiplatelet utility for inhibiting formation both of platelet micro-aggregates and of macro-aggregates of platelets and a long duration of action in humans due to its slow dissociation from human platelets. PMID- 11023247 TI - Quasi-immune response of Penaeus japonicus to penaeid rod-shaped DNA virus (PRDV). AB - A quasi-immune response was demonstrated in kuruma prawn Penaeus japonicus infected naturally or experimentally with PRDV (penaeid rod-shaped DNA virus, also called white spot syndrome virus or WSSV), the causative agent of PAV (penaeid acute viremia). In the first step of this study, natural survivors 4 mo after a PAV outbreak demonstrated 94 % relative percent survival (RPS) upon experimental PRDV challenge. Mortalities after challenge were confirmed by PRDV detection to be due to PAV using a PCR method. In the second step, experimental PAV survivors were produced by intramuscular (IM) injection of PRDV into naive shrimp subsequently reared collectively in a tank (A group) or individually in chamber units (B group). Survival was 41 and 90% in the A and B groups, respectively. A subsequent IM re-challenge of these PRDV survivor groups with PRDV made 32 d after the first challenge revealed a protective response with high RPS of 77 and 64%, respectively. These high survival rates suggested that PAV survivors (natural or experimental) were able to resist PRDV infection and that the resistance was not due to selection of naturally resistant shrimp during a PAV outbreak, but due to enhancement of an immune-like system (quasi-immune response) after exposure to PRDV. No PRDV neutralizing activity was revealed in the serum of the 4 mo natural survivors of the PRDV outbreak. However, it was found in their serum 17 d after they had been experimentally challenged with PRDV. PMID- 11023248 TI - Periodic occurrence of epithelial viral necrosis outbreaks in Penaeus vannamei in Ecuador. AB - Epizootics of an infectious cuticular epithelial necrosis virus (ICENV) occurred in cultured Penaeus vannamei in Ecuadorian shrimp farms from 1994 to 1996. There were few reports of outbreaks during 1997, but in the second half of 1998 epizootics were again reported. Histopathological examination revealed extensive tissue changes and necrosis as described for infections by what others have called Taura syndrome virus (TSV). Infiltration of haemocytes in the cuticular epithelium was also one of the characteristics in the subacute and acute forms of this disease. Electron microscopy of affected tissues demonstrated the presence of a single type of virus particle in the cytoplasm of diseased shrimps from these outbreaks and it corresponded to the published descriptions for TSV. The epizootics of ICENV were periodic in occurrence, and data indicated that they might be related to the oceanographic and climatic variations reported in the eastern Pacific from 1994 to 1998. Using 3 mo rolling averages, a statistically significant negative correlation was found between prevalence of ICENV and temperature but not temperature change. By contrast, there was a statistically significant positive correlation between prevalence of ICENV and salinity change but not salinity value. Although these data do not establish causal relationships, they suggest that laboratory tests should be conducted to determine whether low temperature and upward changes in salinity can increase shrimp susceptibility to infection and mortality by ICENV. PMID- 11023249 TI - Glycogen granules in resting and inflammatory rainbow trout phagocytes--an ultrastructural study. AB - The ultrastructural image of glycogen granules in the cytoplasm of rainbow trout phagocytes in sections stained by the conventional lead or uranyl-lead stains is highly dependent on fixation conditions, the granules being visible only when adequate fixation protocols are used. Morphometry of samples processed for the detection of peroxidase or esterase activities (to specifically label neutrophils and macrophages, respectively), and simultaneously stained for the specific detection of glycogen, showed that inflammatory peritoneal neutrophils were richer in glycogen granules than resting neutrophils. This increase in glycogen content occurs after the migration from the haematopoietic tissues and peripheral blood to the inflamed foci. Glycogen granules could not be found in resting peritoneal macrophages but were found in inflammatory macrophages. The macrophage granules occurred in smaller amounts than in neutrophils, and consisted of granules identical to those of neutrophils together with significantly smaller granules. No evidence for the utilization of glycogen by neutrophils phagocytosing bacteria within the peritoneal cavity was found. PMID- 11023250 TI - Prey, parasites and pathology associated with the mortality of a juvenile gray whale (Eschrichtius robustus) stranded along the northern California coast. AB - An eastern Pacific gray whale (Eschrichtius robustus) stranded off Pelican Point, Tomales Bay, California, USA, was examined for physiological parameters, prey, parasites and associated pathology. The whale was emaciated, and hematological examination revealed an elevation in hematocrit, serum sodium, potassium, electrolyte values and hypoglycemia. Parasites recovered included 5 species, 1 ectoparasite (Cyamus scammoni), and 4 helminths (Anisakis simplex, Ogmogaster antarcticus, Ogmogaster pentalineatus, Bolbosoma balanae) with the latter causing multifocal transmural abscesses. Histological examination indicated severe acute lung congestion, minimal, multifocal, lymphocytic, interstitial myocarditis, and mild hepatocellular and Kupffer cell hemosiderosis. The prey taxa present in the stomach indicated the whale was feeding on hard bottom communities prior to death. PMID- 11023251 TI - Regional patterns in prevalence of principal external diseases of dab Limanda limanda in the North Sea and adjacent areas 1992-1997. AB - The prevalence and spatial distribution of major diseases of dab Limanda limanda in the North Sea and adjacent areas were studied in the summers 1992 to 1997. Areas covered were the North Sea, Irish Sea, northern and northeastern British Waters and the English Channel. The diseases studied were lymphocystis, epidermal hyperplasia/papilloma and skin ulceration. To standardise data, results were analysed for females >15 cm (>3 yr old). Data were subjected to median polish, and additive, extended and additive plus multiplicative models were applied to best account for effects of region and year. Annual differences in disease prevalence were low whilst differences between areas were pronounced. For lymphocystis higher prevalence was observed in the northwestern sector of the North Sea, at the northern tip of Scotland and in an area south of Iceland. Prevalence was low in the Irish Sea, the English Channel and the southern North Sea, and intermediate in the German Bight. For epidermal hyperplasia/papilloma, levels were low at Icelandic stations, in the northern Irish Sea, in the southern North Sea and the English Channel, whilst levels were high in the northwestern part of the North Sea and the German Bight. Elevated levels of skin ulceration were found on the Dogger, at 1 station in the Irish Sea (off Sellafield) and at 1 station to the south of Iceland. Lower levels were detected west of Iceland. Prevalence in all other areas was intermediate. It is concluded that a detailed analysis of available data on disease prevalence and putative causative factors is desirable and, given the good availability of data, would be a promising step forward toward elucidating possible cause and effect relationships between diseases and anthropogenic factors. PMID- 11023253 TI - Infectious diseases in Mazzaella laminarioides (Rhodophyta): estimating the effect of infections on host reproductive potential. AB - Very little is known about the potential effects of endophytic infections on the host in algae, especially in terms of fitness. In this study, we report a first attempt at quantifying the effects of the endophytes Endophyton ramosum (Chlorophyta) and Pleurocapsa sp. (Cyanophyta) on the reproductive potential of their host Mazzaella laminarioides (Rhodophyta). Density of reproductive structures and spore germination rates was compared between infected and non infected fronds. It was found that differences in density of reproductive structures between infected and non-infected fronds were not significant, which suggests that the level of maturity of the host is unaltered by the endophytes. On the other hand, carpospore germination rates significantly decreased when produced by fronds infected by the cyanobacterium Pleurocapsa sp. Thus, in the general context of host fitness, we discuss the fact that only some of the components of the reproductive output of M. laminarioides are negatively affected by the endophytes. PMID- 11023252 TI - Alterations in the biochemistry and ultrastructure of the deep abdominal flexor muscle of the norway lobster Nephrops norvegicus during infection by a parasitic dinoflagellate of the genus Hematodinium. AB - Changes in various biochemical and ultrastructural characteristics of the deep abdominal flexor (DAF) muscles were studied in Norway lobster Nephrops norvegicus (L.) from the Clyde estuary, Scotland, UK, at different stages of infection by a parasitic dinoflagellate of the genus Hematodinium. Abdominal DAF muscles from infected lobsters showed slight, significant increases in total water content, along with greatly depleted glycogen reserves and an altered free amino acid profile. However, protein concentration and composition remained unchanged. Ultrastructurally, parasitic infection of DAF muscle fibres caused alterations in sarcolemmal structure, and localized disruption of myofibrillar bundles around the periphery, but not throughout the centre of the fibres. Overall, the reduction in swimming performance previously reported for N. norvegicus during Hematodinium infection reflect an alteration in carbohydrate supply to the active muscle and some subtle disruption of muscle structure. The altered carbohydrate titre could reflect the Hematodinium parasites acting as a carbohydrate sink in the haemolymph, a disruption of normal tissue glycogenesis, or some alteration in the host's hormonal regulation. These changes could also adversely affect the taste, texture and marketability of infected meat. PMID- 11023254 TI - Trichodina sp. (Ciliophora: Peritrichida) in eel Anguilla anguilla in recirculation systems in denmark: host-parasite relations. AB - Farmed eel cultured in recirculation systems in Denmark were found infected by Trichodina jadranica Raabe, 1958. Associations between parasite abundance and fish size was examined in 2 different production systems. In one system, stocked with relatively well-nourished eels (3 to 31 g), most of the parasites (66%) were found on the dorsal part on the skin and relatively few were found on the gills (approx. 8%). The infection level was significantly positively correlated both with fish weight and length. In the other system, stocked with relatively malnourished small eels (0.5 to 4 g), significantly more parasites (0.06 +/- 0.02 [SD]) were present on fish with a low condition factor than on fish with a higher condition factor (0.13 +/- 0.01 [SD]). In this eel stock the vast majority of the trichodines were found on the gills. PMID- 11023255 TI - Transmission of the haplosporidian parasite MSX Haplosporidium nelsoni to the eastern oyster Crassostrea virginica in an upweller system. AB - The haplosporidian oyster parasite MSX (Multinucleated Sphere X) Haplosporidium nelsoni was transmitted to eastern oysters Crassostrea virginica. Hatchery raised, MSX-free juvenile oysters were placed in upweller tanks. Water to the tanks was filtered through a screen with 1 mm2 openings and originated from the water column overlaying naturally infected oysters beds (MSX prevalence 17 to 57%). MSX was diagnosed by histopathological analysis. MSX-disease (57% prevalence) with increased mortality (19%) was observed 11 wk after the beginning of the exposure and mortality of 80% after 16 wk. The study demonstrates transmission of MSX via water-borne infectious agents capable of passing through a 1 mm filter. PMID- 11023256 TI - Monitoring of infections by protozoa of the genera Nematopsis, Perkinsus and Porospora in the smooth venus clam Callista chione from the North-Western Adriatic Sea (Italy). AB - Marketable smooth venus clams Callista chione from natural banks of Chioggia (Venice) and Goro (Ferrara), North-Western Adriatic Sea (Italy), were examined for protozoan parasites from November 1996 to November 1998. Out of the 375 bivalves examined, 149 (39.7%) were infected by Nematopsis sp. and 325 (86.7%) by Porospora sp. Oocysts of Nematopsis sp. were present with a prevalence that varied from 100% in November 1996 to 5% in June 1998; cystic and naked sporozoites of Porospora sp. were very common, with a prevalence of 100%. Out of the 229 bivalves examined between January and November 1998, 63 (27.5%) were also infected by Perkinsus sp.; the prevalence of Perkinsus sp. varied from 9.1% in January to 50% in February. To our knowledge this is the first report of Porospora sp. and Perkinsus sp. in C. chione. PMID- 11023257 TI - Odontoblast differentiation and tooth morphogenesis. PMID- 11023258 TI - Relaxation vs. cognitively oriented therapies for dental fear. AB - Cognitive therapy has been reported for the treatment of dental phobia, but comparisons with other behavioral treatments are scarce. This study investigated the outcome of two modes of treatment for phobic dental fear. Relaxation and cognitively oriented therapy were compared in a sample of 112 adult fearful dental patients. The patient made questionnaire assessments of background and outcome variables, and the specialist dentist rated successful/non-successful outcome. It was shown that a higher number of patients who received cognitively oriented therapy completed the treatment program, while anxiety was more reduced among patients who received relaxation-oriented therapy. Dropout during the initial phobia therapy with a psychologist was related to lower motivation (willingness to engage in treatment), while failures during dental treatment after the completion of therapy were related to higher levels of general fear and anxiety. A multiple logistic regression model explaining 67% of the variance revealed that the risk of failure was only slightly increased by general fears, while patients with low (below median) motivation ran a 3.6-times-higher risk of dropping out. In conclusion, it was shown that the two treatment methods were both effective in reducing dental phobic reactions. However, while cognitively oriented therapy resulted in a higher number of patients completing therapy, relaxation-oriented treatment generally resulted in a more significant reduction in dental fear as well as in general anxiety and fear. Motivation was found to be a significant predictor of successful treatment outcome. PMID- 11023259 TI - Low unstimulated salivary flow and subjective oral dryness: association with medication, anxiety, depression, and stress. AB - Medication and psychological processes may affect salivary flow and cause subjective oral dryness. The importance of these factors is unclear. The aim of this study was to evaluate the association of medication, anxiety, depression, and stress with unstimulated salivary flow and subjective oral dryness. We studied 1,202 individuals divided into three groups, and controls. Intake of medication was evaluated. Anxiety, depression, and stress were assessed. Unstimulated salivary flow < 0.1 mL/min and subjective oral dryness were significantly associated with age, female gender, intake of psychotropics, anti asthmatics, and diuretics. Unstimulated salivary flow < 0.1 mL/min and no subjective oral dryness were significantly associated with age, intake of antihypertensives, and analgesics. Subjective oral dryness and unstimulated salivary flow > 0.1 mL/min were significantly associated with depression, trait anxiety, perceived stress, state anxiety, female gender, and intake of antihypertensives. Age and medication seemed to play a more important role in individuals with hyposalivation, and female gender and psychological factors in individuals with subjective oral dryness. PMID- 11023260 TI - Measuring the effect of intra-oral implant rehabilitation on health-related quality of life in a randomized controlled clinical trial. AB - The importance of assessing the impact of treatments for chronic conditions on an individual's quality of life has been well-established. In this randomized clinical trial, oral-health-related quality of life, measured with the Oral Health Impact Profile (OHIP), was compared between two groups of edentulous patients. One group (n = 54) received mandibular implant-supported overdentures, and the other group (n = 48) received conventional dentures. Assessments were performed pre-treatment and two months after the prostheses were delivered. The multivariate model showed that implant treatment was significantly associated with lower post-treatment OHIP scores (p = 0.0002), indicating a better quality of life. In addition, pretreatment OHIP scores, treatment allocation, age, sex, and marital status explained 31% of the variation in post-treatment OHIP scores (F = 0.0001). These results suggest that implant treatment provides significant short-term improvement over conventional treatment in oral-health-related quality of life. PMID- 11023261 TI - Prevalence of specific genotypes of Porphyromonas gingivalis fimA and periodontal health status. AB - Porphyromonas gingivalis fimA gene encoding fimbrillin, a subunit of fimbriae, has been classified into 5 genotypes (types I to V) based on their nucleotide sequences. Here, we investigated the relationship between the prevalence of these fimA genotypes and periodontal health status in adults. Dental plaque specimens obtained from 380 periodontally healthy adults and 139 periodontitis patients were analyzed by the PCR method. P. gingivalis was detected in 36.8% of the healthy subjects and in 87.1% of the periodontitis patients. Among the P. gingivalis-positive healthy adults, the most prevalent fimA type was type I (76.1%), followed by type V. In contrast, a majority of the periodontitis patients carried type II fimA organisms (66.1%), followed by type IV. The univariate analysis illustrated that periodontitis was associated with the occurrences of type I fimA (OR 0.16), type II (OR 44.44), type III (1.96), type IV (13.87), and type V (1.40). These findings clearly indicate that there are both disease-associated and non-disease-associated strains of P. gingivalis, and that their infectious traits influencing periodontal health status could be differentiated based on the clonal variation of fimA genes. PMID- 11023262 TI - Expression of the antimicrobial peptide, human beta-defensin 1, in duct cells of minor salivary glands and detection in saliva. AB - The oral cavity is exposed to a variety of environmental insults. Salivary secretions play a critical role in maintaining oral health via innate host defense mechanisms and secretion of secretory IgA. Human beta-defensins (hBD) are antimicrobial peptides that are a component of the innate immune response; they are expressed in epithelia and are proposed to have a role in mucosal defense. hBD-1 mRNA is constitutively expressed in numerous mucosal tissues, including human gingiva and submandibular and parotid glands. Our objective was to detect the expression and localization of hBD-1 peptide in human salivary glands and in saliva. Minor salivary gland tissue was obtained from biopsies of patients with mucoceles (n = 20). hBD-1 peptide was detected by immunohistochemistry; expression was localized to the ductal cells and not the acinar cells of these glands. The peptide was located apically, toward the lumen in the duct cells. Further evaluation showed stronger hBD-1 expression in ducts with periductal inflammation, as indicated by the immunostaining of serial sections with anti CD45 specific for B- and T-lymphocytes. Statistical analysis showed a strong correlation of hBD-1 staining and inflammation. Results of immunolocalization suggest that hBD-1 functions to protect salivary glands from retrograde infection, that expression of the peptide is enhanced in inflamed sites, and that post-transcriptional regulatory mechanisms may be involved in hBD-1 peptide expression. Western immunoblot analysis also detected hBD-1 peptide in unstimulated, whole, acidified saliva from normal volunteers. However, hBD-1 peptide associated with salivary mucin resulted in loss of the detection in a dot immunoblot assay. Association of hBD-1 with salivary mucin may facilitate peptide distribution and adherence to oral surfaces and aid its function within the oral cavity. PMID- 11023263 TI - Cytokine production and bone remodeling in patients wearing overdentures on oral implants. AB - The stability of titanium dental implants is determined by osseointegration. Bone is a dynamic tissue continuously remodeled through resorption and formation, processes controlled by local cytokine production. This study investigated osseotropic cytokine expression in gingival mucosa, in the intraforamina and inferior first molar zones, during rehabilitation with implant-retained overdentures. Specimens were taken from six patients prior to placement of implants in the intraforamina bone; at connection of healing abutments; and 4, 8, and 12 months after prosthetic anchorage. Through semi-quantitative reverse transcriptase polymerase chain-reaction, the following constitutively expressed cytokines were found at first surgical stage: interleukin-1, -6, and -8; small amounts of interleukin-11; stem cell factor; and transforming growth factor beta1, -beta2, and -beta3. From the connection of healing abutments to 12 months after prosthetic anchorage, transforming growth factor-beta1, -beta2, and -beta3 were markedly higher than initial values. Expression of interleukin-6 and -8 decreased 8 months after prosthetic anchorage, while that of interleukin-1 increased at 12 months. In cultured gingival fibroblasts, modulation of cytokine secretion was also time-dependent. Cell culture supernatants influenced osteoclast-like multinucleated cell formation in long-term human marrow culture or osteoblast function, depending on the cytokine profile produced. These results are consistent with functional contributions of cytokines to osseointegration and minimization of posterior edentulous zone bone resorption. PMID- 11023264 TI - Induction of intracellular interleukin-1 beta signals via type II interleukin-1 receptor in human gingival fibroblasts. AB - The type II interleukin-1 receptor (IL-1RII) has been thought to be incapable of transducing signals to cells because of its short intracellular domain, while type I IL-1 receptor (IL-1RI) does transduce signals. Since over-expression of IL 1RII has been demonstrated to inhibit cytokine production in the fibroblastic cell line, it has been proposed to use IL-1RII to prevent IL-1-induced inflammation in connective tissue. In this study, trace amounts of IL-1RII mRNA expression were detected in human gingival fibroblasts (HGFs), which are affected by cytokines in inflammatory periodontal disease. Cloning of the cDNA encoding IL 1RII expressed in HGFs revealed 3 amino acid substitutions in the extracellular domain, when compared with the 408 residues predicted from human B-cells. Over expression of IL-1RII on HGFs by gene transfer down-regulated the expression of IL-1 beta mRNA and IL-6 mRNA in response to IL-1 beta stimulation, while the expression of IL-8 mRNA was not affected. In the IL-1RII-transfected HGFs, phosphorylation of 25- and 74-kDa proteins was up-regulated upon IL-1 beta stimulation in the transfected HGFs. The phosphorylation of these proteins was suppressed by the addition of a neutralizing antibody against IL-1RII. These results suggest that the IL-1RII may regulate HGFs expression of cytokine mRNA upon IL-1 beta stimulation, possibly by altering the IL-1RI-dependent signals. PMID- 11023265 TI - Effects of TGFbeta1 on dental pulp cells in cultured human tooth slices. AB - Transforming growth factor-beta1 (TGF beta1) is a potent modulator of tissue repair in various tissues. To analyze its role during human dental repair, we used thick-sliced teeth cultured as described previously (Magloire et al., 1996). The supply of TGF beta1 to the pulp tissue was accomplished by means of a small tube glued onto the dentin. We show that this device allowed the growth factor to diffuse locally through dentinal tubules and to bind to the cells present in the coronal pulp opposite the TGF beta1-delivery tube. The tube was filled with 20 ng/mL TGF beta1, and slices were cultured for 4 days. Results show a preferential accumulation of cells in the odontoblastic and subodontoblastic layers in the vicinity of the tube. Cell proliferation increased in the subodontoblastic layer and in the underlying pulp, and BrdU-positive cells were abundant around the blood vessels. TGF beta1 induced type I collagen production by the odontoblastic/subodontoblastic/pulp cells in the stimulated zone, as demonstrated by in situ hybridization. These results suggest that TGF beta1 could be directly involved in the regulation of cell proliferation, migration, and extracellular matrix production in the human dental pulp and eventually in the repair process occurring after tooth injury. PMID- 11023266 TI - Cloning, characterization, and expression analysis of mouse enamelysin. AB - Enamelysin is a recently isolated member of the matrix metalloproteinase (MMP) family of extracellular matrix (ECM)-degrading enzymes. Here we describe the isolation and characterization of the mouse enamelysin cDNA. Expression of mouse enamelysin was detectable only in ameloblasts and odontoblasts of developing teeth. Characterization of mouse enamelysin demonstrated that it is highly conserved in both its sequence content and pattern of expression relative to the porcine, human, and bovine homologues previously described. PMID- 11023267 TI - Meal pattern analysis in response to temporomandibular joint inflammation in the rat. AB - Inflammation of the temporomandibular joint (TMJ) can alter behavioral responses such as food intake and mobilize stress hormones. The hypothesis of this study was that food intake and diurnal corticosterone analysis can be used as indicators of adjuvant-induced TMJ inflammation. Groups of rats received adjuvant or no injections at the beginning of the resting (AM) or activity (PM) phase. Forty-eight hours (early) or 6 weeks (late) after adjuvant injection, plasma corticosterone was assayed and food intake was recorded. Food intake was suppressed up to 4 days post-injection. As expected, the non-injected group showed low AM and high PM corticosterone. AM corticosterone was elevated, but PM corticosterone was attenuated in both early- and late-stage-injected rats. A computerized pair-fed experiment showed that adjuvant-induced hypophagia did not alter corticosterone levels. Meal pattern analysis revealed decreased food intake due to a decrease in the number of meals taken. Notably, meal size remained the same but meal duration increased. This model demonstrated that food intake and stress hormone analysis could be used as indicators for sequelae of adjuvant induced TMJ inflammation. PMID- 11023268 TI - Expression of mRNAs encoding for alpha and beta integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts. AB - The biological mechanisms of tooth movement result from the cellular responses of connective tissues to exogenous mechanical forces. Among these responses, the degradation of the extracellular matrix takes place, but the identification of the molecular basis as well as the components implicated in this degradation are poorly understood. To contribute to this identification, we subjected human fibroblasts obtained from the periodontal ligament (PDLs) and from the gingiva (HGFs) to a continuous stretch to quantify the mRNAs encoding for various metalloproteinases (MMPs), their tissue inhibitors (TIMPs), and alpha and beta integrin subunits. Both cell lines reacted by inducing the expression of the mRNAs encoding for MMP-1, MMP-2, TIMP-1, and TIMP-2, while other mRNAs did not vary (MT1-MMP, TIMP-3) or were not expressed (MMP-9). PDLs expressed selectively the mRNAs encoding for alpha4 and alphav, with no difference measurable under stretching, while the mRNAs encoding for alpha6 and beta1 were increased and the one encoding for alpha5 was decreased. HGFs increased the mRNAs encoding for alpha2, alpha6, beta1, and beta3 and decreased the one encoding for alpha3. Analysis of our data indicated that stretched HGFs and PDLs induced the same pattern of mRNAs encoding for MMPs and TIMPs but differed for those encoding various integrin subunits, known to act as protein receptors in mechanotransduction. PMID- 11023269 TI - Local and chronic application of PTH accelerates tooth movement in rats. AB - We previously reported that whereas systemic continuous infusion of parathyroid hormone (PTH) accelerated orthodontic tooth movement, systemic but intermittent injection of PTH did not increase the rate of tooth movement. Analysis of these data suggested that continuous administration of PTH could be applicable for orthodontic therapy. In the present study, we investigated whether local and chronic application of PTH(1-34) would accelerate orthodontic tooth movement. To increase the residence time of PTH in the injected area, we used methylcellulose (MC) gel (2% W/V) for a slow-release formulation of PTH. MC gel containing PTH (PTH-MC) continuously released biologically active PTH into the acceptor medium for more than 72 hrs in vitro. When male rats received a local injection of PTH MC into the subperiosteum in the mesio-palatal region of the maxillary first molar (M1) every other day, M1 movement, which was mesially drawn by an orthodontic coil spring attached to the maxillary incisors, was accelerated in a dose-dependent manner. PTH-MC injection at 1 microg/400 g body weight caused a 1.6-fold increase in the rate of tooth movement. The acceleration of tooth movement by PTH-MC injection was marked on days 6, 9, and 12. Local injection of PTH dissolved in saline without MC did not significantly accelerate tooth movement on day 6 or later. Histological examination revealed active osteoclastic bone resorption and a widened periodontal space on the compression side of the periodontal tissue in the PTH-MC-injected rats. These results suggest that local injection of PTH in a slow-release formulation is applicable to orthodontic therapy. PMID- 11023270 TI - Effects of low-energy CO2 laser irradiation and the organic matrix on inhibition of enamel demineralization. AB - In the past two decades, accumulated evidence has clearly demonstrated the inhibitory effects of laser irradiation on enamel demineralization, but the exact mechanisms of these effects remain unclear. The purpose of this study was to investigate the effects of low-energy CO2 laser irradiation on demineralization of both normal human enamel and human enamel with its organic matrix removed. Twenty-four human molars were collected, cleaned, and cut into two halves. One half of each tooth was randomly selected and its lipid and protein content extracted. The other half of each tooth was used as the matched control. Each tooth half had two window areas. All the left windows were treated with a low energy laser irradiation, whereas the right windows served as the non-laser controls. After caries-like lesion formation in a pH-cycling environment, microradiographs of tooth sections were taken for quantification of demineralization. The mean mineral losses (with standard deviation) of the enamel control, the lased enamel, the non-organic enamel control, and the lased non organic enamel subgroups were 3955 (1191), 52(49), 4565(1311), and 1191 (940), respectively. A factorial ANOVA showed significant effects of laser irradiation (p = 0.0001), organic matrix (p = 0.0125), and the laser-organic matrix interaction (p = 0.0377). The laser irradiation resulted in a greater than 98% reduction in mineral loss, but the laser effect dropped to about 70% when the organic matrix in the enamel was removed. The results suggest that clinically applicable CO2 laser irradiation may cause an almost complete inhibition of enamel demineralization. PMID- 11023271 TI - Controversies simmering in the dentin field. PMID- 11023272 TI - Forty-four years of dental research: role of education, teachers, research, and teaching. PMID- 11023273 TI - Cytokine profiles of T-lymphocytes from gingival tissues with pathological pocketing. AB - Periodontal disease is an infection in which destruction occurs at sites remote from the infection, resulting in pathological pocketing. Intervening between the infection and the destruction is a dense mononuclear inflammatory infiltrate. It has been suggested that this infiltrate might have characteristics and the destructive potential of Th1-type T lymphocytes. To ascertain the nature of the infiltrates we investigated the expression of mRNA for IL-2, IL-5, and IFN-gamma by gingival mononuclear cells (GMC) from healthy (n = 8) or adult periodontitis (AP) patients (n = 25) by using cytokine-specific reverse transcription/polymerase-chain-reaction (RT-PCR). GMC, as obtained from patients' tissues, expressed IL-2, IFN-gamma, or IL-5 mRNA. Significantly higher proportions of GMC from AP patients expressed IL-2 and IFN-gamma mRNA than did those from healthy subjects. IFN-gamma was the most consistent cytokine message detected. In other experiments, gingival T-lymphocytes (n = 12) and CD4+ and CD8+ gingival T-lymphocytes (n = 16) were isolated from gingival tissues removed surgically from AP patients. AP gingival T-lymphocytes expressed mRNA for IL-2, IFN-gamma, or IL-6 prior to stimulation. After stimulation with Con A, the cells significantly up-regulated IL-5 and IL-6 message expression. Both CD4+ and CD8+ gingival T-lymphocytes expressed IFN-gamma, IL-5, and some IL-2. This cumulative cytokine profile observed in these experiments is consistent with the predominance of Th1-type cells in pathological tissues and with Th2-type cells, which can also be present, being up-regulated under appropriate stimulation. Importantly, CD4+ and CD8+ lymphocytes were shown to express T1- and T2-type cytokine message, emphasizing the potential for CD8+ T-lymphocytes to participate in periodontal disease pathology. PMID- 11023274 TI - Time-dependent decrease and seasonal variation of the porosity of recently erupted sound dental enamel in vivo. AB - Recently erupted teeth are more sensitive to dental caries than teeth that have remained free from caries lesions for a few years after eruption. It has been hypothesized that this may be ascribed to differences in enamel porosity. The objective of the present work was to assess the time-dependence of electrical conductance, as an indication of porosity, of occlusal enamel in recently erupted permanent molar teeth. Fifty children aged 5-15 years of age, receiving regular dental care at six-month intervals, participated in the study when their first molar teeth (in 5- to 6-year-olds) or their second molar teeth (in 11- to 15-year olds) had not been exposed to the oral environment for more than six months. On the first semi-annual check-up after eruption of a first or second molar, baseline diagnostic measurements, i.e., visual inspection and electrical conductance measurements (ECMs), were made at three or four pre-defined sites in the fissures. Subsequently, data collection was repeated every six months for three years. Sites were excluded from ECMs when a caries lesion was observed at a site by visual inspection. After 36 months, there were 257 sites in teeth considered sound upon visual examination. The ECM values of these sound sites showed a clear decrease with time after the first examination. The conductance decreased on average from 0.13 (Momega)(-1) to 0.059 (Momega)(-1). Since the conductance is assumed to be proportional to the porosity of the enamel, the latter decreased by a factor of 2.2 over the 36-month period. The results furthermore indicated a higher conductance for maxillary than for mandibular enamel. Almost all sites in that sample appeared to be in teeth that were observed for the first time during the months September to December. Electrical conductance, and therefore enamel porosity, of the sites showed a periodic variation with season of observation: In the fall, the conductance was higher than in the spring. This implies that a dentist should be more prudent in the fall than in the spring when indicating restorative treatment of an incipient caries lesion. PMID- 11023275 TI - Association of salivary flow rates with maximal bite force. AB - Mean salivary secretion and bite force decrease with advancing age. Previous studies have shown that salivary flow rates are influenced by mastication. In the present study, we examined the relationship between salivary flow rates and maximal bite force in a community-based sample of men and women 35 years of age or older. Salivary flow rates for unstimulated whole and unstimulated submandibular/sublingual (SMSL) saliva as well as citrate-stimulated parotid and SMSL saliva were measured in 399 subjects. Bite force was assessed with a bilateral force transducer. Pearson correlation analysis yielded significant positive correlations between bite force and flow rates for unstimulated whole saliva (r = 0.24, p < 0.0001), stimulated parotid saliva (r = 0.13, p < 0.03), unstimulated SMSL (r = 0.14, p < 0.0001), and stimulated SMSL (r = 0.16, p < 0.003). When adjusted for age and gender, the partial correlations between bite force and salivary flow rates remained significant for unstimulated whole saliva (r = 0.10, p < 0.05), stimulated parotid saliva (r = 0.13, p < 0.02), and stimulated SMSL saliva (r = 0.14, p < 0.006). Subjects were divided into four groups based on their maximal bite force score (low, medium low, medium high, and high). For each saliva type, the flow rate of the high-bite-force group was significantly greater than that of the low-bite-force group as well as that of the medium-high-bite-force group. These results confirm an age-related decrease in bite force and salivary flow rates and show that, regardless of age or gender, bite force is correlated with salivary flow. PMID- 11023276 TI - Mandibular helical axis pathways during mastication. AB - Condylar and incisor trajectories are often used for the study of mandibular movements. Condylar trajectories, however, depend on the location of the reference point and can be interpreted erroneously. In contrast, the helical axis analysis yields an unequivocal description of rigid body kinematics. The aim of this study was to analyze the mandibular helical axis during mastication. Seven subjects without signs and symptoms of craniomandibular disorders and with class I occlusion were recorded by means of the opto-electronic system Jaws-3D during unilateral mastication of bread cubes (2-cm side). The helical axis was computed every 14 ms with a rotation threshold of 1 . Parameters describing its spatial orientation and position relative to the condyles were calculated. The helical axis changed orientation and position more pronouncedly during the closing than during the opening phases of mastication. The orientation varied significantly from beginning to end of closing but not of opening, indicating less fluctuation of the helical axis on opening than on closing. Also, the distance dCP between helical axis and reference condylar point varied more significantly (p < 0.05) on the working than on the balancing side: On the working side, dCP decreased during both opening and closing, whereas on the balancing side, dCP increased only for closing. Furthermore, the helical axis pathway often showed a bowing ventrally to the balancing condyle, indicating that, during closing, the balancing condyle still translated backward while essentially only rotation occurred around the working condyle. Thus, the helical axis changed its position and orientation continuously during mastication. PMID- 11023278 TI - H+-induced tension development in demineralized dentin matrix. AB - Recent atomic force microscopy studies have shown that acid-etching dentin causes the surface to recede. The purpose of this work was to test the hypothesis that acidic solutions can cause contraction of demineralized dentin matrix. Small beams of dentin were cut from extracted human third molars. The central region of each specimen was completely demineralized in EDTA. Specimens held in a tester were immersed in acidic solutions, and the amount of tension was recorded. Test variables included gauge length, cross-sectional area, pre-strain, and pH. The results showed that immersion of demineralized dentin in acidic solutions caused tension that was directly related both to H+ concentration at pH < 2 and to pre strain. The contractile stress development (ca. 0.2-0.4 MPa) was sufficient to cause a collapse of demineralized dentin matrix. The mechanism for this effect probably involves H+-induced conformation changes in the collagen matrix. PMID- 11023277 TI - No heritability of temporomandibular joint signs and symptoms. AB - The causes of temporomandibular joint (TMJ)-related signs and symptoms are largely unknown. We tested the hypotheses that these signs and symptoms, as well as oral parafunctional habits, are substantially heritable. Questionnaire and clinical data were collected from 494 twins, including pairs of reared-apart and reared-together monozygotic (MZ) and dizygotic (DZ) twins. A history of joint area pain, joint noises, and clenching and grinding habits was scored as present or absent. Twenty-nine percent of the population experienced at least one sign or symptom. Nearly one-quarter of subjects clenched or ground their teeth, and 8.7% reported a history of joint-area pain. Pain was associated with clenching, grinding, and joint noises. MZ twins were no more similar than DZ twins for any outcome, suggesting that genetic factors do not influence these traits in the population. Reared-together MZ twins were no more similar than reared-apart MZ twins, suggesting a negligible effect of the family environment on these outcomes. Environmental factors unique to each twin appeared to be the major determinants of variation in this population. PMID- 11023279 TI - Optical scattering power for characterization of mineral loss. AB - Mineral loss in early caries cannot be measured without invasive procedures. To quantify mineral loss without sectioning the tooth, one must determine the optical scattering of the enamel. Using enamel white-spot lesions, we hypothesize that the optical scattering power (Sp) of the demineralized enamel would provide a quantitative estimate of mineral loss. Enamel slabs were demineralized to produce artificial white spots. The data were acquired by means of a Charge Coupled Device (CCD) camera and image-processing software. For the purpose of comparison, mineral loss (deltaZ) of the demineralized samples was determined by the use of a microhardness approach after the samples were sectioned. The scattering power correlated well with deltaZ (r2 = 0.82). In contrast, simple reflectance of the demineralized samples correlated poorly with deltaZ (r2 = 0.22). The validity of using scattering power to measure demineralization has been confirmed by a three-dimensional Monte Carlo Simulation. PMID- 11023280 TI - High-temperature X-ray diffraction measurement of sanidine thermal expansion. AB - Dental porcelains that are designed to be fused to PFM (porcelain-fused-to-metal) alloys are formulated by their manufacturers to be closely matched in thermal expansion to these alloys. The high thermal expansion of the mineral leucite has been exploited to regulate porcelain expansion. Leucite, however, has been observed to convert to the sanidine polymorph of feldspar during certain heat treatments within the normal firing range of dental porcelain. The effects of this conversion on porcelain thermal expansion and porcelain-metal thermal compatibility have been uncertain, due to the paucity of published data on the thermal expansion of sanidine. The purpose of this study was to measure the thermal expansion of sanidine by high-temperature X-ray diffraction over the temperature range in which thermal mismatch stresses can develop in porcelain fused-to-metal restorations, i.e., from room temperature to 700 degrees C. The lattice parameters a, b, c, and beta were determined from the d-spacings and hkl values of multiple reflections by means of a least-squares iteration. The dependence of each lattice parameter on temperature was determined via analysis of variance, and the coefficient of thermal expansion, alpha, was obtained from this analysis. The lattice parameters of sanidine at room temperature were determined to be: a = 0.8524 +/- 0.0015 nm, b = 1.3020 +/- 0.0004 nm, c = 0.7165 +/- 0.0002 nm, and beta = 116.02 degrees +/- 0.01 degree (mean +/- 95% confidence interval). The linear thermal expansion coefficient, a, over the range from room temperature to 700 degrees C was determined to be 4.1 x 10(-6) K(-1) +/- 0.6 x 10(6) K(-1) (mean +/- 95% confidence interval). Because the coefficient of thermal expansion for sanidine is substantially lower than that of leucite (the effective linear thermal coefficient of thermal expansion of leucite over the range of 25 degrees to 700 degrees C is 28 x 10(-6) K(-1)), the conversion of leucite to sanidine during porcelain heat treatments would produce a detrimental lowering of the porcelain thermal expansion. PMID- 11023282 TI - Prostaglandin E2 enhances bradykinin-evoked iCGRP release in bovine dental pulp. AB - Mediators produced during inflammation are responsible for hyperalgesia and expression of neurotransmitters and receptors in the nervous system. The production of bradykinin (BK) and the prostaglandins (PGs) may regulate initiation of pain. This study tested the hypothesis that BK and prostaglandin E2 (PGE2) have a positive interaction in evoking neurosecretion of immunoreactive calcitonin gene-related peptide (iCGRP). Bovine dental pulp was prepared and stimulated by the superfusion method with BK alone and in combination with PGE2. Kinin receptor antagonists to bradykinin-evoked release of iCGRP were also tested. Also tested was the hypothesis that dental pulp contains either the B1 or B2 or both BK receptors. Results showed that PGE2 enhanced BK-evoked iCGRP release by more than 50%. Western immunoblots revealed detectable B2 receptor protein with no detectable B1 receptor protein. We conclude that BK evokes iCGRP release from bovine dental pulp which is enhanced by a positive interaction with PGE2. Neurosecretion is evoked from isolated terminals of dental pulp fibers via the bradykinin B2 receptor-dependent mechanism. PMID- 11023281 TI - The role of vascular endothelial growth factor in human dental pulp cells: induction of chemotaxis, proliferation, and differentiation and activation of the AP-1-dependent signaling pathway. AB - Vascular endothelial growth factor (VEGF) is a potent mitogen in endothelial cells, but little is known about its activity in other cell types. To clarify the role of VEGF in human dental pulp cells and pulp tissue, we investigated the effects of VEGF on the chemotaxis, proliferation, and differentiation of human dental pulp cells. VEGF induced a strong chemotactic response in human dental pulp cells in a dose-dependent manner. VEGF also marginally enhanced the proliferation of human dental pulp cells and induced an increase in alkaline phosphatase in human dental pulp cells. However, these effects of VEGF were not observed in reference to human skin fibroblasts. Analyses by the reverse transcription/polymerase-chain-reaction method and flow cytometry showed that the mRNAs of two VEGF receptors, fins-like tyrosine kinase and kinase insert domain containing receptor, were expressed in human dental pulp cells, whereas only fms like tyrosine kinase mRNA was expressed in human skin fibroblasts. VEGF induced the activation of activator protein 1 (AP-1) and c-fos mRNA expression in human dental pulp cells. The AP-1 inhibitor curcumin strongly inhibited VEGF-induced alkaline phosphatase production in human dental pulp cells. In addition, VEGF antisense oligonucleotide suppressed the production of VEGF and alkaline phosphatase in human dental pulp cells. These results suggest that VEGF produced by human dental pulp cells acts directly upon human dental pulp cells in an autocrine manner, and may promote the chemotaxis, proliferation, and/or differentiation of human dental pulp cells via the utilization of kinase insert domain-containing receptor and in part through AP-1 by increasing c-fos. PMID- 11023283 TI - Immunohistochemical detection and distribution of enamelysin (MMP-20) in human odontogenic tumors. AB - Enamelysin is a tooth-specific protease that was initially isolated from porcine enamel organ and subsequently from human odontoblasts. Since this protease is thought to play important roles in tooth development, the evaluation of enamelysin in odontogenic tumors may aid our understanding of the histogenesis and cell differentiation of such lesions. A monoclonal antibody (203-1C7) was generated against synthesized human enamelysin oligopeptide and was used to assess the immunolocalization of enamelysin in healthy developing tooth germs and various types of odontogenic lesions. In tooth germs, enamelysin expression was detected only in the secretory enamel. Thus, 203-1C7 may serve as an enamel specific marker in the late stage of enamel matrix development and calcification. In odontogenic lesions, strong enamelysin staining was demonstrated in the immature enamel matrix of ameloblastic fibro-odontomas and odontomas. Furthermore, enamelysin was also detected in globular amyloid masses and calcified foci in calcifying epithelial odontogenic tumors, hyaline droplets, small and large mineralized areas in adenomatoid odontogenic tumors, and a portion of ghost cells in calcifying odontogenic cysts. Positive reactivity was also observed in selected tumor cells in some of these tumors. No intracellular staining for enamelysin was detected in ameloblastomas or the ameloblastic portion of ameloblastic fibro-odontomas. Also, enamelysin was not detected in dentin, dysplastic dentinoid hyaline matrices, and cementum that were present within the tumors examined. Thus, taken together, our results suggest that the enamelysin-specific monoclonal antibody (203-1C7) may be utilized as a marker of early enamel development and that enamelysin may be involved in the pathogenesis of specific odontogenic tumors. PMID- 11023284 TI - High glucose suppresses cathepsin activity in periodontal-ligament-derived fibroblastic cells. AB - The accumulation of extracellular matrices and integrins by high glucose has been reported in relation to diabetic complications. We previously reported that PDL cells expressed a higher amount of VLA-5 when cultured in high-glucose (4500 mg/L) medium than those cultured in low-glucose (1100 mg/L) medium. In this study, we aimed to address (1) whether this effect was mediated by the transcriptional level of the gene or the degradative level of the protein, and (2) whether this effect was mediated by TGF-beta. The results indicated that the level of mRNA encoding alpha5 integrin did not change in PDL cells regardless of the concentration of glucose. Alternatively, high glucose suppressed cathepsin B+L activity. Additionally, the level of mRNA encoding TGF-beta was not affected by high glucose, nor did an anti-TGF-beta neutralizing antibody have an effect on the expression of beta5 gene or cathepsin activity. Therefore, the effects of high glucose appeared to be mediated by impaired protein degradation, but not by autocrine TGF-beta. PMID- 11023285 TI - Myosin isoform composition of the human medial and lateral pterygoid muscles. AB - The medial and lateral pterygoid muscles are different in structure as well as in function. The medial pterygoid muscle is concentrically active during jaw closing, and the superior head of the lateral pterygoid muscle is eccentrically active during jaw closing, while its inferior head is concentrically active during jaw opening. Architecturally, the medial pterygoid can deliver higher forces than the lateral pterygoid. We investigated whether these differences are reflected in the myosin heavy-chain (MyHC) composition and the fiber cross sectional area (f-csa) of these muscles. The pterygoid muscles from eight cadavers were investigated by means of monoclonal antibodies against different isoforms of MyHC. The proportions of pure MyHC type I fibers did not differ significantly among the muscles (32% in medial pterygoid, 34% in superior head, and 36% in the inferior head of the lateral pterygoid), nor did the total proportions of pure MyHC type IIA and IIX fibers (16% in medial pterygoid, 26% in the superior head, and 19% in the inferior head of the lateral pterygoid). The mean f-csa of type I fibers was 1315 microm2, which did not differ significantly among the muscles, and was significantly larger than the f-csa of type IIA fibers. The relative proportions of hybrid fibers, which expressed more than one MyHC isoform, were 52% in the medial pterygoid, 40% in the superior head, and 45% in the inferior head of the lateral pterygoid and did not differ significantly among the muscles. The most abundant hybrid fiber types found were fibers expressing MyHCs-cardiac alpha+IIA and MyHCs-cardiac alpha+I+IIA. Significant regional differences were found in the proportions of MyHC type I fibers in the medial pterygoid and in the inferior head of the lateral pterygoid. Although the form and function of the muscles are different, we conclude that this is not reflected in their myosin isoform composition. PMID- 11023287 TI - Effect at one school of clerkship curriculum and examination modifications on students' USMLE step 2 scores. PMID- 11023286 TI - Jaw-tongue reflex: afferents, central pathways, and synaptic potentials in hypoglossal motoneurons in the cat. AB - The tongue position is reflexively controlled by the jaw position (the jaw-tongue reflex). The purpose of this study was to clarify the mechanism of this reflex in terms of afferents, central pathways, and synaptic potentials in hypoglossal motoneurons in the cat. Intracellular recordings from hypoglossal motoneurons revealed that electrical stimulation of the temporalis muscle nerve evoked excitatory and inhibitory post-synaptic potentials in hypoglossal motoneurons. The threshold of temporalis muscle nerve stimulation for evoking the synaptic potentials was higher than 2.0 times the nerve threshold. The amplitude of the potentials increased with stimulus intensity up to 5.0 times the nerve threshold. Punctate light pressure applied to the temporalis muscle induced a tonic depolarizing potential in hypoglossal motoneurons on which action potentials as well as depolarizing synaptic activation noise were superimposed. On the other hand, electrical stimulation of the temporalis muscle during jaw-opening could slightly inhibit the electromyographic activities in the genioglossus and styloglossus muscles. Lesions including the Probst's tract at the level caudal to the trigeminal motor nucleus abolished both excitation and inhibition in hypoglossal motoneurons induced by tonic depression of the lower jaw, but exerted no effects on either the tonic stretch reflex or the trigemino-hypoglossal reflex. In contrast, lesions including the trigeminal spinal tract produced no changes in either excitation or inhibition of hypoglossal motoneurons induced by temporalis muscle afferents, whereas the excitation of hypoglossal motoneurons was abolished by the lesions. We conclude that the group II muscle spindle afferents from the temporalis muscle are primarily responsible for evoking the jaw-tongue reflex. PMID- 11023288 TI - Nationwide perspective on the use of a formal ethics curriculum during critical care fellowship training. PMID- 11023289 TI - Knowledge acquisition in a one-month infectious diseases rotation. PMID- 11023290 TI - Women's health programs in academic health centers. PMID- 11023291 TI - Blood purification by immunoadsorption. PMID- 11023292 TI - A thymine base is missed out of the published nucleotide sequence dat of the anti thrombin gene. PMID- 11023293 TI - The e19a2 bcr/abl breakpoint in acute lymphoblastic leukaemia. PMID- 11023294 TI - Plasma levels of vascular endothelial growth factor (VEGF) in haematological cancers. PMID- 11023295 TI - Hepatitis C virus infection associated with B-cell non-Hodgkin's lymphoma in Hungarian patients. PMID- 11023296 TI - Compound heterozygosity for triplicated alpha-globin gene and (- -(SEA)) alpha globin gene deletion: implication for thalassaemia screening. PMID- 11023297 TI - Cardiac hyperacute rejection: a new look at an old problem. PMID- 11023298 TI - Case 2: cardiogenic shock due to acute vascular rejection in a heart transplant recipient. PMID- 11023299 TI - Cautionary note: complex (dys) function of the serotonin transporter. PMID- 11023300 TI - Web alert. Pattern formation and developmental mechanisms. PMID- 11023301 TI - Pattern formation and developmental mechanisms. The cell biological basis of inductive signaling. PMID- 11023302 TI - Evo-devo of feathers and scales: building complex epithelial appendages. PMID- 11023303 TI - Differentiation and gene regulation. Web alert. PMID- 11023304 TI - Ending genital mutilation. Male genital mutilation in any society is surely abhorrent too. PMID- 11023305 TI - Speed of treatment affects outcome in anaphylaxis. PMID- 11023306 TI - Why is speculation so awful? PMID- 11023307 TI - All antihistamines cross blood-brain barrier. PMID- 11023308 TI - More public education and more intubationists will prevent prehospital deaths. PMID- 11023309 TI - The NHS: last act of a Greek tragedy? Government that puts money into redressing inequalities is worthy of support. PMID- 11023310 TI - Students learn infection control on the job. PMID- 11023311 TI - The NHS: last act of a Greek tragedy? Creativity is not valued in public sector. PMID- 11023312 TI - The NHS: last act of a Greek tragedy? NHS should be abolished. PMID- 11023313 TI - Information needs of patients with cancer. Patients are frightened and their information needs fluctuate. PMID- 11023314 TI - Information needs of patients with cancer. Similar study had similar findings. PMID- 11023315 TI - Information needs of patients with cancer. Patients' perspectives may vary. PMID- 11023316 TI - Breaking down barriers for refugee doctors. Doctors can qualify in the United Kingdom. PMID- 11023317 TI - Breaking down barriers for refugee doctors. Secure statutory finding is needed. PMID- 11023318 TI - National service framework for coronary heart disease. Ambiguities need to be clarified. PMID- 11023320 TI - National service framework for coronary heart disease. Many operators and facilities will not meet standards set out in framework. PMID- 11023319 TI - National service framework for coronary heart disease. Target of lowering cholesterol by 30% needs to be justified. PMID- 11023321 TI - Brain stem death. Healthcare workers have difficulty accepting current management. PMID- 11023322 TI - Brain stem death. Patients' interests should predominate. PMID- 11023323 TI - Doctors who are unsympathetic to colleagues who are psychologically vulnerable. PMID- 11023324 TI - Assessing aggression in psychiatric inpatients. Assessing aggression can be risky. PMID- 11023325 TI - Assessing aggression in psychiatric inpatients. Study should have been controlled. PMID- 11023326 TI - In China, suicide in young women is a problem too. PMID- 11023328 TI - Optimal age for starting lipid lowering treatment. It is more efficient to screen and treat elderly people. PMID- 11023327 TI - Optimal age for starting lipid lowering treatment. A 10 year risk of 30% should be used. PMID- 11023329 TI - Optimal age for starting lipid lowering treatment. Adjusted date do not justify a lower optimal age. PMID- 11023331 TI - Action on Cataracts should influence surgical training. PMID- 11023330 TI - Anticoagulation for patients with atrial fibrillation. Editorial may have overinterpreted data. PMID- 11023332 TI - Anticoagulation for patients with atrial fibrillation. Warfarin should be given for up to one year after successful cardioversion. PMID- 11023333 TI - Photodynamic therapy: a new concept in medical treatment. AB - A new concept in the therapy of both neoplastic and non-neoplastic diseases is discussed in this article. Photodynamic therapy (PDT) involves light activation, in the presence of molecular oxygen, of certain dyes that are taken up by the target tissue. These dyes are termed photosensitizers. The mechanism of interaction of the photosensitizers and light is discussed, along with the effects produced in the target tissue. The present status of clinical PDT is discussed along with the newer photosensitizers being used and their clinical roles. Despite the promising results from earlier clinical trials of PDT, considerable additional work is needed to bring this new modality of treatment into modern clinical practice. Improvements in the area of light source delivery, light dosimetry and the computation of models of treatment are necessary to standardize treatments and ensure proper treatment delivery. Finally, quality assurance issues in the treatment process should be introduced. PMID- 11023334 TI - Surveillance of viral infections in donated blood, England and Wales, 1999. PMID- 11023335 TI - Inter- and intra-individual variability of ground reaction forces during sit-to stand with principal component analysis Medical Engineering & Physics 21 (1999): 235-240. PMID- 11023337 TI - Annual scientific meeting of the Association of Paediatric Anaesthetists of Great Britain and Ireland. PMID- 11023336 TI - Microtubule bundling. PMID- 11023338 TI - Guess what! Angiosarcoma of the face and scalp. PMID- 11023339 TI - Guess what! Superficial migratory thrombophlebitis. Thromboangiitis obliterans (Buerger's disease). PMID- 11023340 TI - Guess what! Diagnosis and comments: Birt-Hogg-Dube syndrome. PMID- 11023341 TI - The cost and time to develop new drugs has increased dramatically. PMID- 11023342 TI - Optimizing the science of drug development: opportunities for better candidate selection and accelerated evaluation in humans. PMID- 11023343 TI - Novel survivin-related members of the inhibitor of apoptosis (IAP) family. PMID- 11023344 TI - [Malaria in Benghazi City, Libyan Arab Jamahiriya]. PMID- 11023345 TI - 24th International Symposium on GH and Growth Factors in Endocrinology and Metabolism. Antwerp, October 1997. Proceedings and abstracts. PMID- 11023346 TI - Holger W. Jannasch (1927-1998): "small is powerful". PMID- 11023347 TI - Eladio Vinuela (1937-1999), the molecular biology pioneer in Spain. PMID- 11023348 TI - [HIV and antiretroviral therapy: in every 3rd patient a virus rebound takes place]. PMID- 11023349 TI - [Systemic lupus erythematosus: the prognosis has improved]. PMID- 11023351 TI - An eruption of silicosis. PMID- 11023350 TI - Women's environmental health. PMID- 11023352 TI - Bridging the gap in border health. PMID- 11023353 TI - The origin of pyruvate: NADP+ oxidoreductase in mitochondria of Euglena gracilis. PMID- 11023354 TI - Disulfide bridges of ergtoxin, a member of a new sub-family of peptide blockers of the ether-a-go-go-related K+ channel. PMID- 11023355 TI - Pesticide residues variability and acute dietary risk assessment. Conference proceedings. 1-3 December 1998, York, United Kingdom. PMID- 11023356 TI - New endoscopic strategies offer middle ground for treating GERD. PMID- 11023357 TI - Solving the African enigma: parasites may have their place. PMID- 11023358 TI - Image of the month. Cause and mechanism of constipation in a 58-year-old woman. PMID- 11023359 TI - Biographical sketch. Bernard of the duct of Bernard. PMID- 11023360 TI - Is small-caliber esophagogastroduodenoscopy more tolerable than conventional one in unsedated patients? PMID- 11023361 TI - Polypectomy in chronic colitis. PMID- 11023362 TI - What is the risk of biliary pancreatitis in patients with gallstones? PMID- 11023363 TI - CD4+ T cells in cerulein-induced pancreatitis. PMID- 11023364 TI - High incidence of oral lichen planus in an HCV hyperendemic area. PMID- 11023365 TI - Drug allergies among patients with borderline personality symptomatology. PMID- 11023366 TI - Olanzapine-induced bilateral pedal edema: a case report. PMID- 11023367 TI - Syphilis in psychiatric inpatients: prevalence, treatment and implications. PMID- 11023368 TI - [Tardive dyskinesia and the therapeutic agents]. PMID- 11023370 TI - Is the WHO classification suitable for skin lymphoma? PMID- 11023369 TI - [Therapeutic administration of trace elements and manganese poisoning]. PMID- 11023371 TI - European stillbirth proportion and Chernobyl. PMID- 11023372 TI - [Managed care in Germany: comparison of two inpatient case management approaches]. PMID- 11023373 TI - [From prevention to aftercare]. PMID- 11023374 TI - [Evidence-based medicine and quality assurance]. PMID- 11023375 TI - [When may the physician reveal information that is protected by confidentiality?]. PMID- 11023376 TI - ["Much delighted thus the eyes peacefully closed..." Thoughts on the 250th anniversary of the death of Johann Sebastian Bach]. PMID- 11023377 TI - On the pressure gradient boundary condition for the contact of two biphasic cartilage layers. PMID- 11023378 TI - Comments on 'Elastic modulus and hardness of cortical and trabecular bone lamellae measured by nanoindentation in the human femur'. PMID- 11023379 TI - Herlitz junctional epidermolysis bullosa: novel and recurrent mutations in the LAMB3 gene and the population carrier frequency. AB - Herlitz junctional epidermolysis bullosa is a heritable bullous disease caused by mutations found primarily in the b3 chain of laminin 5 (LAMB3). In this study, we examined the LAMB3 gene for mutations in 22 Herlitz junctional epidermolysis bullosa families, and identified 15 distinct mutations, eight of them previously unreported, bringing the total number of distinct Herlitz junctional epidermolysis bullosa mutations in LAMB3 to 35. Examination of the mutation database revealed several recurrent mutations that have been reported, as well as six previously unreported. All recurrent mutations may be readily detected by polymerase chain reaction of genomic DNA and restriction endonuclease digestion. Mutation screening and prenatal diagnosis of families at risk may be expedited by molecular testing for these recurrent mutations prior to screening the entire gene. Finally, the U.S. population carrier risk for Herlitz junctional epidermolysis bullosa and all variants of junctional epidermolysis bullosa was calculated to be one in 781 and one in 350, respectively, while the overall epidermolysis bullosa carrier frequency was calculated to be one in 113. These data allow accurate testing, counseling, and risk calculation for nuclear families, as well as extended family members at risk for junctional epidermolysis bullosa. PMID- 11023380 TI - Common morality versus specified principlism: reply to Richardson. AB - In his article 'Specifying, balancing and interpreting bioethical principles' (Richardson, 2000), Henry Richardson claims that the two dominant theories in bioethics--principlism, put forward by Beauchamp and Childress in Principles of Bioethics, and common morality, put forward by Gert, Culver and Clouser in Bioethics: A Return to Fundamentals--are deficient because they employ balancing rather than specification to resolve disputes between principles or rules. We show that, contrary to Richardson's claim, the major problem with principlism, either the original version or the specified principlism of Richardson, is that it conceives of morality as being composed of free-standing principles, rather than as common morality conceives it, as being a complete public system, composed of rules, ideals, morally relevant features, and a procedure for determining when a rule can be justifiably violated. PMID- 11023381 TI - Reply to Strong on principlism and casuistry. PMID- 11023382 TI - The quality of surgical information on the Internet. PMID- 11023383 TI - The Student National Medical Association: another successful year. PMID- 11023385 TI - From the Centers for Disease Control and Prevention. Update: newborn screening for sickle cell disease--California, Illinois, and New York, 1998. PMID- 11023384 TI - Re: andropause: a misnomer for a true clinical entity. PMID- 11023386 TI - From the Centers for Disease Control and Prevention. State-specific cholesterol screening trends--United States, 1991-1999. PMID- 11023387 TI - From the Centers for Disease Control and Prevention. Publication of Surgeon General's report on smoking and health. PMID- 11023388 TI - From the Centers of Disease Control and Prevention. CDC statement on study results of product containing nonoxynol-9. PMID- 11023389 TI - JAMA patient page. Taking care of your gums. PMID- 11023390 TI - [Effects of transcutaneous heart-lung support on left ventricular dysfunction in septicemia]. PMID- 11023391 TI - Nefazodone, psychotherapy, and their combination for chronic depression. PMID- 11023392 TI - Nefazodone, psychotherapy, and their combination for chronic depression. PMID- 11023393 TI - Nefazodone, psychotherapy, and their combination for chronic depression. PMID- 11023394 TI - Nefazodone, psychotherapy, and their combination for chronic depression. PMID- 11023395 TI - Philadelphia chromosome-positive acute lymphoblastic leukemia in children. PMID- 11023396 TI - Philadelphia chromosome-positive acute lymphoblastic leukemia in children. PMID- 11023397 TI - Treatment of the nephrotic syndrome with etanercept in patients with the tumor necrosis factor receptor-associated periodic syndrome. PMID- 11023398 TI - The volume of primary angioplasty procedures and survival after acute myocardial infarction. PMID- 11023399 TI - Health advice for travelers. PMID- 11023400 TI - Health advice for travelers. PMID- 11023401 TI - Malaria in a Trinidadian man. PMID- 11023402 TI - Malaria in a Trinidadian man. PMID- 11023403 TI - Systemic absorption of food dye in patients with sepsis. PMID- 11023404 TI - Parasite-altered behaviour: is the effect of Toxoplasma gondii on Rattus norvegicus specific? AB - The effect of Toxoplasma gondii on rat behaviour was assessed in a colony of 36 free-ranging wild/laboratory hybrid rats allowed to compete freely for food and mates in a (100 m2) outdoor naturalistic enclosure. T. gondii infection had no effect on social status or mating success, both the product of costly and competitive activities. However, the propensity to explore novel stimuli in their environment was higher in infected than uninfected individuals. These results are consistent with the hypothesis that T. gondii only affects the behavioural traits which selectively benefit the parasite, rather than causing a general alteration of rat behaviour. PMID- 11023405 TI - Identification and characterization of three Encephalitozoon cuniculi strains. AB - Microsporidia are increasingly recognized as causing opportunistic infections in immunocompromised individuals. Encephalitozoon cuniculi is probably the most studied mammalian microsporidian that infects insects and mammals, including man. In this study, 8 E. cuniculi isolates were compared and were found to fall into 3 strains. Strain type I includes the rabbit type isolate, as well as isolates from an additional rabbit, a dwarf rabbit, and a mouse. Strain type II includes 2 murine isolates and strain type III includes 2 isolates obtained from domestic dogs. By SDS-PAGE, the 3 strains differ primarily in the molecular weight range of 54-59 kDa where strain type I displays an apparent broad singlet at 57 kDa, strain type II displays an apparent doublet at 54 and 58 kDa, and strain type III displays an apparent broad band at 59 kDa. Antigenic differences were detected in the molecular weight regions of 54-58 kDa as well as 28-40 kDa by Western blot immunodetection using murine antisera raised against E. cuniculi, Encephalitozoon hellem, and the Encephalitozoon-like Septata intestinalis. Polymerase chain reaction (PCR) products containing only small subunit rDNA sequences from the different E. cuniculi isolates formed homoduplexes whereas PCR products containing intergenic rRNA gene sequences formed heteroduplexes in mobility shift analyses. Fok I digestion of the PCR products containing the intergenic rRNA gene region resulted in unique restriction fragment length polymorphism patterns, and DNA sequencing demonstrated that in the intergenic spacer region, the sequence 5' GTTT-3' was repeated 3 times in strain type I, twice in strain type II, and 4 times in strain type III. This study indicates that there exist at least 3 E. cuniculi strains which may become important in the epidemiology of human E. cuniculi infections. Furthermore, as additional E. cuniculi isolates are characterized, these strains will be named or reclassified once the criteria for taxonomy and phylogenetic tree construction for microsporidia become better defined. PMID- 11023406 TI - A Giardia duodenalis gene encoding a protein with multiple repeats of a toxin homologue. AB - A gene, CRP136, from the intestinal protozoan parasite Giardia duodenalis, expressed at a high level in a metronidazole-resistant line, encodes a 136 kDa protein with 23 copies of a 40 amino acid repeat. The protein is cysteine-rich and has the typical membrane-spanning region and CXXC amino acid motifs of a family of Giardia cysteine-rich surface proteins (CRSPs). The repeat unit in CRP136, shares 57% homology with the gene encoding the precursor of the sarafotoxins, a group of snake toxins from the burrowing adder known to cause symptoms similar to those of humans acutely infected with Giardia. The sarafotoxins are low molecular weight sulphydryl cross-linked peptides which are proteolytically cleaved from a precursor polyprotein. CRP136 has homology over the entire length of the sarafotoxin precursor, and the repeats are of the same length. Thus CRP136 represents the first evidence for a potential Giardia toxin. The genomic copy number of CRP136 appears to be the same in both the parent and drug-resistant lines and expression of this gene, and at least one other, is associated with a conserved partial duplication, but not amplification, of one chromosome. PMID- 11023407 TI - Gene flow and cross-mating in Plasmodium falciparum in households in a Tanzanian village. AB - The diversity of the genes encoding 2 merozoite surface proteins (MSP-1 and MSP 2) of Plasmodium falciparum has been examined in parasites infecting members of 4 households in a village in Tanzania. The polymerase chain reaction (PCR) was used to characterize allelic variants of these genes by the sizes and sequences of regions of tandemly repeated bases in each gene. In each household extensive polymorphism was detected among parasites in the inhabitants and in infected mosquitoes caught in their houses. Similar frequencies of the alleles of these genes were observed in all households. Capture-recapture data indicated that both Anopheles gambiae and A. funestus freely dispersed among households in the hamlet. The results confirm that cross-mating and gene flow occur extensively among the parasites, and are discussed within the context of spatial clustering of natural populations of P. falciparum. PMID- 11023408 TI - Effects of endotoxin and dexamethasone on cerebral malaria in mice. AB - CBA/T6 and DBA/2J mice inoculated with Plasmodium berghei ANKA (PbA) develop cerebral involvement 6-8 days post-inoculation, from which the CBA mice almost invariably die and the DBA mice recover. Dexamethasone (DXM; 80 mg/kg) given to inoculated CBA mice twice, on day 3 and again within 48 h, reduced the cerebral symptoms and prevented death from cerebral malaria. Plasma tumour necrosis factor (TNF) levels, which increased at the time of the cerebral symptoms, were also reduced in these DXM-treated mice. Intravenously administered Evans Blue, a dye which binds to albumin, diffused extensively across the blood-brain barrier only during the period of cerebral symptoms, in proportion to the severity of the cerebral symptoms and the disease. In PbA-infected CBA mice, cerebral symptoms and the amounts of Evans Blue diffusing into the brain tissue were both reduced by DXM treatment, but only if the steroid was given on day 3 and again within 48 h. Endotoxin injected intravascularly into PbA-infected DBA mice after day 5 resulted in an exaggeration of cerebral symptoms and death between days 6 and 9. Plasma TNF and the amount of Evans Blue in the brain parenchyma increased above normal levels in these mice. Endotoxin injections had only minor effects on the severity of the cerebral symptoms in PbA-infected CBA mice and did not cause the animals to die sooner. PMID- 11023409 TI - The entry of Theileria parva merozoites into bovine erythrocytes occurs by a process similar to sporozoite invasion of lymphocytes. AB - The entry of Theileria parva merozoites into bovine erythrocytes in vivo is described and compared to sporozoite invasion of lymphocytes. Merozoites make initial contact with erythrocytes with any part of their surface and invasion of the host cell does not require the re-orientation of the apical end of the merozoite towards the surface of the erythrocyte. After the initial attachment the merozoite and host cell membranes form a continual close junction with the two apposed membranes separated by a 6-8 nm gap containing moderately dense material. The progressive circumferential 'zippering' of these closely apposed membranes leads to the movement of the parasite into the erythrocyte. The newly internalized merozoite which is completely surrounded by the erythrocyte plasma membrane escapes from this enclosing membrane by a process involving the discharge of at least the rhoptries; whether the merozoite also contain other types of secretory organelles (e.g. micronemes, microspheres or dense bodies) remains to be determined. Morphologically, the events involved in merozoite invasion of erythrocytes are almost identical to the process of sporozoite invasion of lymphocytes but differ significantly from the entry process of the invasive stages of other Apicomplexan parasites. PMID- 11023410 TI - Optimization of the in vitro feeding of Rhipicephalus appendiculatus nymphae for the transmission of Theileria parva. AB - An apparatus for artificial feeding of Rhipicephalus appendiculatus nymphae was modified to improve feeding performance. Heparinized blood was supplied above a treated artificial membrane while the ticks attached below on its undersurface. The feeding apparatus was incubated at 37 degrees C in an atmosphere of 3% CO2 concentration and a relative humidity of 75-80%. Under these conditions, 91% of the engorged nymphae attained a mean weight of 6-11 mg, and an average of 93% of those nymphae moulted into adults. When this system was used to feed nymphal ticks on blood infected with Theileria parva piroplasms, the mean prevalence of infection in the resultant female and male ticks was 86% and 54%, respectively. The feeding performance and T. parva infection levels were comparable to those of nymphal ticks fed on the blood donor cattle. The apparatus used in this study has potential for modification to suit the artificial feeding needs of other species of ixodid ticks and for use in investigations to examine other tick/pathogen relationships. PMID- 11023411 TI - Transcriptional regulation of Schistosoma mansoni calreticulin: possible role of AP-1. AB - Little is known about the regulation and control of Schistosoma mansoni gene expression. In order to study such mechanisms a gene reporter expression vector construction, under the control of a promoter region derived from the S. mansoni calreticulin gene was used to transfect the human Jurkat T cell line. The promoter region contains potential TATA and CAAT boxes as well as an AP-1 core element. We show here that transcriptional factors of eucaryotic cells may induce a gene reporter activity under the control of a S. mansoni promoter region. Treatment of stably transfected cells with N-acetyl cysteine (NAC), a well characterized antioxidant which counteracts the effects of reactive oxygen intermediates, enhanced the AP-1 dependent transactivation. This effect was abolished when the SmCaR promoter region was deleted in the AP-1 site. Electrophoretic Mobility Shift Assays showed that the AP-1 sequence of S. mansoni bound to both S. mansoni extracts and in nuclear extracts from Jurkat cells, thus explaining possible activation of AP-1 by NAC. Finally treatment of S. mansoni schistosomula and adult worms with NAC induced an increased synthesis of calreticulin protein suggesting a possible role of redox mechanisms in the regulation of a calreticulin gene transcription process in S. mansoni. PMID- 11023412 TI - A female-specific cDNA sequence of Schistosoma mansoni encoding a mucin-like protein that is expressed in the epithelial cells of the reproductive duct. AB - Seven cDNA clones of Schistosoma mansoni containing the C-terminal part of the deduced sequence of a mucin-like protein have been identified. The protein contains 28% threonines, 20% serines, and has a pI of 3.4. On Northern blots of RNA of adult worms, the cDNA clones detect 2 transcripts of 1.65 and 4.2 kb which are expressed only in female worms. The tissue of gene expression, as revealed by in situ hybridization, is the epithelium surrounding the female reproduction duct proximal to its entrance into the ootype. Accumulation of N-glycosylation sites suggests that the protein, like other mucins, might form a protective layer, coating the lining of the duct. Regarding its acidic pI, we hypothesize a role in preventing premature egg-shell formation. This is the first female-specifically transcribed sequence, hitherto known in S. mansoni that is not expressed in the vitellaria. PMID- 11023413 TI - Observations on worm population dynamics in calves naturally infected with Schistosoma mattheei. AB - The evolution of faecal egg output, worm burdens and tissue egg counts in young calves was monitored during the first year of natural exposure to Schistosoma mattheei infection on a Zambian farm. According to the duration of their stay on the farm, these calves were classified into 2 groups of 14 temporary tracers (TT calves) which were introduced on a 2-monthly basis for residential periods of 2 months, and 12 permanent tracers (PT calves) introduced either at the beginning of the experiment (Group A) or 2 months later (Group B) and gradually removed after residential periods of 2, 4, 6, 8, 10 and 12 months on the farm. Worm counts in the TT calves showed that infection occurred throughout the year on the farm and that levels of infection acquired during each period of 8 weeks correlated well with the respective infected snail densities observed at the main transmission site. Marked differences in worm population dynamics were recorded between the 2 groups of PT calves. In Group B animals which apparently were initially exposed to heavy transmission, according to the results from TT calves, much higher worm counts and greater susceptibility to reinfection were observed than in Group A animals initially exposed to lighter exposure. These results suggest that the development of resistance to natural infection with S. mattheei may depend on the initial exposure to the parasite. Low initial exposures may lead to resistance whereas high initial exposures may result in decreased immune responses resulting in susceptibility to infection. PMID- 11023414 TI - Effects of phenothiazine neuroleptic drugs on the microtubular-membrane complex in bloodstream forms of Trypanosoma brucei. AB - African trypanosomes are parasitic protozoa causing sleeping sickness in humans and related diseases in domestic animals against which no entirely satisfactory forms of chemotherapy are yet available. It was previously shown that related species of trypanosomes, as well as procyclic (insect) forms of Trypanosoma brucei are extremely sensitive to the action of phenothiazine neuroleptic drugs in vitro. In this work, we have carried out a more detailed investigation of the effects of thioridazine, one of the most potent neuroleptic phenothiazine drugs known, on the morphology of the infective bloodstream forms of T. brucei, with particular reference to the parasite's prominent pellicular membrane complex. Our data show that this drug induces rapid changes in cell shape that appear to involve some reorganization of the microtubular membrane skeleton, but does not affect the structural integrity of the microtubular complex. Another early consequence of drug action involved damage to nuclear and cytoplasmic membranes and the appearance of tubular arrays of coated membrane within the flagellar pocket. It was also revealed that the drug induces a rapid release of the variant specific glycoprotein (VSG) which makes up the surface coat protecting bloodstream forms of the parasite against the host immune system. Our evidence suggests that this release of VSG involves cleavage of the protein's glycosyl phosphatidylinositol (GPI) membrane anchor by endogenous GPI-specific phospholipase C, probably as a consequence of minor damage to the parasite plasma membrane induced by the drug. PMID- 11023415 TI - The FMRFamide-like neuropeptide AF2 is present in the parasitic nematode Haemonchus contortus. AB - Peptides belonging to the FMRFamide family are widely distributed amongst invertebrates. We report here on the isolation of the FMRFamide neuropeptide AF2 (Lys-His-Glu-Tyr-Leu-Arg-Phe-NH2) from the parasitic nematode Haemonchus contortus. Immunocytochemical techniques showed that FMRFamide-like material was distributed in several regions of these organisms including nerve cords and cell bodies of the central nervous system. AF2 was isolated using a method that employed 6 steps of reverse-phase HPLC. The concentration of AF2 in this organism was approximately 30 pmol/g of nematode. PMID- 11023416 TI - A quick-frozen, freeze-fracture and deep-etched study of the cuticle of adult forms of Strongyloides venezuelensis (Nematoda). AB - The cuticle of adult forms of Strongyloides venezuelensis was studied by routine transmission electron microscopy, conventional freeze-fracture and also using quick-freeze and deep-etch techniques. In routine thin sections the cuticle of S. venezuelensis comprises 7 layers: epicuticle, outer cortical, inner cortical, external medial, internal medial, fibrous and basal. Observation of replicas of specimens fractured across the thickness of the body wall, revealed at the epicuticle an ordered array of particles accompanying the cuticular annulations. At the level of the cortical and medial layers we observed few scattered particles embedded in an amorphous matrix without a particular arrangement. The fibrous layer was represented by several parallel lines of ordered particles of similar size. In tangentially fractured specimens, the epicuticle cleaves readily exposing 2 faces, one exhibiting intramembranous particles without any particular arrangement, immersed in a smooth matrix (P face), and the other showing depressions and very few particles (E face). In replicas of fractures submitted to etching, we observed at the level of the cortical, medial fibrous and basal layers an interconnecting fibrous and globous structure which was organized in a different direction at the fibrous layer. The association of freeze-fracture to deep-etch technique revealed the internal structural organization of the cuticle layers showing details that were not seen before using conventional freeze fracture technique. PMID- 11023417 TI - Computer model of the maintenance and selection of genetic heterogeneity in polygamous helminths. AB - A stochastic simulation model of the transmission and maintenance of genetic heterogeneity in the absence and presence of external selection pressures is presented for polygamous intestinal helminths such as Ascaris. The model assumes that the density distribution of the adult parasites is highly aggregated and that density-dependent effects on fecundity are important. The model gives rise to stable infection rates in the host. Where the parasite population contains genetic heterogeneity, with the exception of stochastic fluctuations which models genetic drift, the ratio of the different alleles remained constant over extended periods of time. This result contrasts with that of an earlier analytical model (Anderson, R.M., May, M.R. & Gutpa S. (1989) Parasitology 99, S59-S79), in which uneven mating probabilities for the different combinations of worm possible in a host was postulated to inevitably lead to fixation of the most abundant allele. New results suggest that in spite of the restricted choice of mating available to a worm in the confines of a host, selection pressure always leads to enrichment of the parasites carrying resistant alleles. PMID- 11023418 TI - Genetic heterogeneity in helminths - a reply. PMID- 11023419 TI - [The diagnosis of the pharynx. Its anatomy--pathology--pathophysiology]. PMID- 11023420 TI - A timely reminder. PMID- 11023421 TI - [Treatment of cows that are not yet in estrus]. PMID- 11023422 TI - [Review of the origin, the continuation and the end of the Dutch Association of Women Veterinarians]. PMID- 11023423 TI - [Taking responsibility]. PMID- 11023424 TI - [Let me introduce Polidore Calon, meat cutter in Ijzendijke. Reaction to, "Laparoscopic ovariectomy in the standing mare."]. PMID- 11023425 TI - [Spring Days Committee set for 2000 World Congress. All veterinarians for companion animals go to Amsterdam]. PMID- 11023426 TI - Pain behavior in animals entirely subtle. Lizette Hardie: "The owner expects us to do something about it.". PMID- 11023427 TI - [Veterinary editors gather in the RAI. European Association of Veterinary Editors meeting on the topic of the Internet]. PMID- 11023428 TI - Fritz Haber: December 9, 1868-January 29, 1934. AB - Fritz Haber (1868-1934) was a German physical chemist. Nobel laureate and foreign member of the US National Academy of Sciences. His greatest accomplishment in science was the development of a practical method to prepare nitrogen from air (nitrogen fixation or Haber-Bosch process). While working on the toxicity of war gases. he formulated 'Haber's rule', also known as C x T= constant in order to characterize the toxicity of an inhalant. Between 1919 and 1933. he was one of the leading figures in revitalizing science in Germany. At his institute in Berlin worked such luminaries as Albert Einstein, Lise Meitner and Otto Hahn. His last paper described what became known as the Haber-Weiss reaction. After his death he was for a long time forgotten by the Nazis because he was Jewish and after World War II by the Allies because of his work on war gases in World War I. And yet he was one of the truly great modern scientists. not only because of his science, but also because of the role he played in science politics and policies. PMID- 11023429 TI - [Prof. Vladimir Aleksiev, 1879-1948]. PMID- 11023430 TI - Proceedings of the International Symposium on Viral Hepatitis Beyond the Millennium. 9-11 December 1999, Leuven, Belgium. PMID- 11023431 TI - Improvement of chronic active hepatitis C in chronically infected chimpanzees after therapeutic vaccination with the HCV E1 protein. PMID- 11023432 TI - Proceedings of the XIVth International Symposium on Arterial Chemoreception. June 24-28, 1999, Philadelphia, Pennsylvania, USA. PMID- 11023433 TI - Case 1. Phyllodes (phylloides or cystosarcoma phyllodes) tumor: wide local excision is the preferred method of treatment. PMID- 11023434 TI - Proceedings of the 10th International Congress on Echocardiography: Clinical Cardiology. February 1999, Rome, Italy. PMID- 11023435 TI - Minimizing the progression of kidney disease. Introduction. PMID- 11023436 TI - Three generations of patients with lupus erythematosus and hereditary angioedema. PMID- 11023437 TI - Chronic fatigue syndrome. PMID- 11023438 TI - Mixed connective-tissue disease presenting with transient complete heart block. PMID- 11023439 TI - Metformin and late gastrointestinal complications. PMID- 11023440 TI - Rhabdomyolysis due to combination therapy with cerivastatin and gemfibrozil. PMID- 11023441 TI - Nanoliter liquid metering in microchannels using hydrophobic patterns. AB - Nanoliter-sized liquid drops can be accurately metered inside hydrophilic microchannels using a combination of hydrophobic surface treatment and air pressure. The technique involves spontaneously filling the microchannels up to a hydrophobic region and splitting a liquid drop by injecting air through a hydrophobic side channel. The hydrophobic regions are fabricated by using a patterned metal mask on a substrate. The patterned substrate is immersed in an isooctane solution containing 1H,1H,2H,2H-per-fluorodecyltrichlorosilane to form hydrophobic patches on the exposed surface. Stripping the metal mask leaves the hydrophobic patches and restores the hydrophilic substrate surface. Precise and accurate liquid volumes, ranging from 0.5 to 125 nanoliters, have been metered using this technique. Theoretical predictions of the pressure needed to meter drops compare well with the experimental values. PMID- 11023443 TI - An important source of error in asbestos-related lung cancer estimates. PMID- 11023442 TI - Epidemiological evidence on the carcinogenicity of silica: factors in scientific judgement. PMID- 11023444 TI - Are SHV beta-lactamases universal in Klebsiella pneumoniae? PMID- 11023445 TI - Linkage of vat(E) and erm(B) in streptogamin-resistant Enterococcus faecium isolates from Europe. PMID- 11023446 TI - The image of nursing. PMID- 11023447 TI - Newsworthy or not worthy. PMID- 11023448 TI - Finding information abut glaucoma on the Internet. PMID- 11023449 TI - Serum skeletal troponin I in inflammatory muscle disease: relation to creatine kinase, CKMB and cardiac troponin I. PMID- 11023450 TI - Enlarged spleen detected by abdominal ultrasonography in patients with RA. PMID- 11023451 TI - Ultrasound measurement of brain atrophy in multiple sclerosis. PMID- 11023452 TI - The prevalence and incidence of hepatitis C virus infections among dialysis patients in the Netherlands: a nationwide prospective study. AB - A nationwide prospective survey on hepatitis C virus (HCV) infections among dialysis patients in The Netherlands was performed. Patients were recruited from 34 dialysis centers and were tested for antibodies and HCV RNA in 1995 and 1997. Seronegative serum samples were analyzed by reverse-transcriptase polymerase chain reaction in pools. HCV-RNA-positive serum samples were genotyped and were partly sequenced. In the first and second rounds, 67 (2.9%) of 2281 and 76 (3.4%) of 2286 patients were HCV positive, respectively. Of 960 patients with paired serum samples, 35 were HCV positive in both rounds, and 9 HCV-positive cases were newly identified in the second round. The incidence of HCV infection was 0.5 per 100 dialysis years. Phylogenetic analysis revealed clustered sequences that indicated nosocomial transmission. Sixty percent of HCV infections, however, can be attributed to 4 interdependent risk factors (i.e., hemodialysis before 1992, kidney transplantation before 1994, and birth or dialysis in a foreign country). In conclusion, the prevalence of HCV infections in The Netherlands does not decline, and transmission within dialysis units continues. Adequate screening of HCV infections and strict enforcement of universal infection control practices are required. PMID- 11023453 TI - Prospective evaluation of infection with hepatitis G virus in relation to hepatocellular carcinoma in Shanghai, China. AB - The health consequences of hepatitis G virus (HGV) infection in humans remain to be determined. Inconsistent results of HGV and the risk of hepatocellular carcinoma (HCC) have been found in retrospective case-control studies. No prospective studies have addressed this issue. A prospective study including 18,244 men in Shanghai, China, was conducted to assess the role of HGV infection in HCC development. Serum HGV RNA and hepatitis B surface antigen (HBsAg) were assessed in 127 HCC case patients and 660 matched control subjects drawn from the cohort. Four patients with HCC (3.1%) and 21 control subjects (3.2%) were positive for serum HGV RNA. The presence of HGV RNA in prediagnostic serum was not associated with the risk of HCC (odds ratio, 1.0; 95% confidence interval, 0.3-2.9). No association between HGV RNA positivity and HCC risk was observed in either HBsAg-positive carriers or noncarriers. It is concluded that HGV infection plays no role in HCC development in this high-risk population. PMID- 11023454 TI - Herpes simplex virus DNA vaccine efficacy: effect of glycoprotein D plasmid constructs. AB - The impact of vaccination with plasmid DNA encoding full-length glycoprotein D (gD) from herpes simplex virus (HSV) type 2 (gD2), secreted gD2, or cytosolic gD2 was evaluated in mice and guinea pigs. Immunization with plasmids encoding full length gD2 or secreted gD2 produced high antibody levels, whereas immunization with DNA encoding cytosolic gD2 resulted in significantly lower antibody titers in both species (P<.001). Vaccination with DNA encoding full-length or secreted gD2 significantly reduced acute disease in mice and guinea pigs (both P<.001) and subsequent recurrent disease in guinea pigs (P<.05). In guinea pigs, immunization with DNA encoding cytosolic gD2 did not protect from acute or recurrent disease, whereas in mice it did protect, but not as well as DNA encoding full-length or secreted gD2. None of the vaccines resulted in improved virus clearance from the inoculation site, and none significantly reduced recurrent disease when used as a therapeutic vaccine in HSV-2-infected guinea pigs. PMID- 11023455 TI - Plasmid DNA-expressed secreted and nonsecreted forms of herpes simplex virus glycoprotein D2 induce different types of immune responses. AB - Herpes simplex viruses (HSVs) are significant pathogens and major targets of vaccine development. Several attempts have been made to develop prophylactic and therapeutic vaccines for HSV types 1 and 2. Although these vaccines elicit strong humoral responses, the overall impact on pathology has been disappointing. An effective vaccine for HSV must induce both humoral and cellular immune responses. DNA vaccines are ideal candidates for HSV vaccines because they induce both types of immune responses. This study showed that the type of immune response generated by immunization with DNA vaccines is modulated by expression of various forms of an antigen, each with a different cellular localization. Expression of cell associated forms of HSV-2 glycoprotein D (gD) induces primarily a Th1 response, whereas expression of secreted gD results in a Th2 response. Immunization with plasmids expressing different forms of the antigen may increase the efficacy of a vaccine. PMID- 11023456 TI - Tissue distribution and variant characterization of human herpesvirus (HHV)-6: increased prevalence of HHV-6A in patients with multiple sclerosis. AB - Human herpesvirus (HHV)-6 has been associated with the pathogenesis of multiple sclerosis (MS) on the basis of serologic, molecular, and histopathologic studies. This study sought to determine the distribution of HHV-6 in different MS body fluids, including serum, saliva, urine, and peripheral blood lymphocytes. The study results extend the observation of an increased frequency of HHV-6 DNA in serum of patients with MS to the unique detection of viral sequences in urine of a subset of patients with MS. Moreover, the HHV-6 identified in these cell-free compartments was predominantly the HHV-6A variant, which has been reported to be neurotropic. These results support the hypothesis that HHV-6 may contribute to the MS disease process. PMID- 11023458 TI - Cellular immunity to human immunodeficiency virus type 1 (HIV-1) clades: relevance to HIV-1 vaccine trials in Uganda. AB - The first prophylactic human immunodeficiency virus type 1 (HIV-1) vaccine trial in Africa, with a clade B immunogen, is currently under way in Uganda, in a region where clades A and D are endemic. The use of a B clade vaccine is based on anticipated cross-recognition of endemic strains of HIV-1 in Uganda, but, in fact, little is known about the cytotoxic T lymphocyte (CTL) responses in that region. Seventeen HIV-1-infected volunteers from Kampala, Uganda, were studied to determine the immune responses elicited by natural infection with local HIV-1 strains. Despite the presence of broad cross-clade recognition, the CTL responses to the infecting viral clade were highest in most people. Recognition of nonendemic clade B antigens was similar to that of the coendemic local clade, and, in some instances, cross-recognition of clade B was greater. Nevertheless, the degree of cross-clade cellular responses we observed lends justification to the use of clade B-based immunogens in the current phase 1 vaccine trial in Uganda. PMID- 11023459 TI - Two double-blinded, randomized, comparative trials of 4 human immunodeficiency virus type 1 (HIV-1) envelope vaccines in HIV-1-infected individuals across a spectrum of disease severity: AIDS Clinical Trials Groups 209 and 214. AB - The potential role of human immunodeficiency virus type 1 (HIV-1)-specific immune responses in controlling viral replication in vivo has stimulated interest in enhancing virus-specific immunity by vaccinating infected individuals with HIV-1 or its components. These studies were undertaken to define patient populations most likely to respond to vaccination, with the induction of novel HIV-1-specific cellular immune responses, and to compare the safety and immunogenicity of several candidate recombinant HIV-1 envelope vaccines and adjuvants. New lymphoproliferative responses (LPRs) developed in <30% of vaccine recipients. LPRs were elicited primarily in study participants with a CD4 cell count >350 cells/mm(3) and were usually strain restricted. Responders tended to be more likely than nonresponders to have an undetectable level of HIV-1 RNA at baseline (P=.067). Induction of new cellular immune responses by HIV-1 envelope vaccines is a function of the immunologic stage of disease and baseline plasma HIV-1 RNA level and exhibits considerable vaccine strain specificity. PMID- 11023460 TI - Mucosal and systemic immune activation is present in human immunodeficiency virus exposed seronegative women. AB - Immune parameters were analyzed in peripheral blood mononuclear cells (PBMC) and cervical mucosa biopsy specimens of human immunodeficiency virus (HIV) seronegative women sexually exposed to HIV (exposed seronegative [ESN]), HIV infected women, and healthy women without HIV exposure. HIV was not detected in PBMC or cervical mucosa biopsy specimens of ESN women. However, interleukin (IL) 6, IL-10, IL-12, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha and -beta mRNA were elevated in PBMC and cervical mucosa biopsy specimens of ESN and HIV-infected women; CCR5 and CXCR4 mRNA were augmented in cervical mucosa biopsy specimens, but not in PBMC, of ESN and HIV-infected women; HIV-specific IFN-gamma-secreting cells were detected in vaginal washes of ESN and HIV-infected women; and phenotypic alterations were present in PBMC of ESN women. These results suggest that active HIV infection is not required for T cell activation; immune alterations occur in women in whom HIV infection cannot be detected virologically or clinically. PMID- 11023461 TI - Randomized study of saquinavir with ritonavir or nelfinavir together with delavirdine, adefovir, or both in human immunodeficiency virus-infected adults with virologic failure on indinavir: AIDS Clinical Trials Group Study 359. AB - This study compared antiretroviral activity among 6 "salvage" therapy regimens. The study was a prospective, randomized, 2x3 factorial, multicenter study of the AIDS Clinical Trials Group. The study enrolled 277 human immunodeficiency virus (HIV)-infected patients naive to nonnucleoside analogues who had taken indinavir >6 months. The patients had 2000-200,000 HIV RNA copies/mL. Patients received saquinavir with ritonavir or nelfinavir together with delavirdine and/or adefovir and were followed for safety and antiretroviral response between baseline and week 16. At week 16, 30% (77/254) of patients had 4 weeks. Administration of acellular pertussis vaccine in these age groups could prevent this substantial morbidity. PMID- 11023465 TI - The emergence of Streptococcus pneumoniae resistant to macrolide antimicrobial agents: a 6-year population-based assessment. AB - From 1994 through 1999, the available isolates (4148 isolates) from active population-based surveillance of invasive pneumococcal disease in metropolitan Atlanta were serotyped and were tested for antimicrobial susceptibility. Macrolide-resistant isolates were studied for the presence of ermAM (a ribosomal methylase gene), mefE (a macrolide efflux gene), and tetM (the class M tetracycline resistance gene). Macrolide resistance increased from 16% of all invasive isolates in 1994 to 32% in 1999. Of the macrolide-resistant pneumococcal isolates studied, 99% contained genomic copies of mefE or ermAM. Isolates with ermAM were mainly serotypes 6B, 23F, 14, or 19F and contained tetM; mefE associated isolates were predominantly serotypes 14, 6A, or 19F, and most did not contain tetM. The frequency of the ermAM-mediated phenotype in invasive Streptococcus pneumoniae remained stable over the 6-year surveillance. However, the mefE-mediated phenotype increased from 9% in 1994 to 26% of all isolates in 1999 and was noted in new serotypes. By 1999, 93% of the mefE-containing strains had minimum inhibitory concentrations >/=8 microgram/mL. Dissemination of the mefE determinant accounted for the rapid increase in the rate of macrolide resistance in our S. pneumoniae population. PMID- 11023466 TI - Pharmacologic interference with NF-kappaB activation attenuates central nervous system complications in experimental Pneumococcal meningitis. AB - This study assessed the effects of 2 different inhibitors of NF-kappaB activation on central nervous system complications and clinical symptoms in an advanced stage of experimental pneumococcal meningitis: the calpain inhibitor I N-acetyl leucinyl-leucinyl-norleucinal (ALLN), which interferes with IkappaB proteolysis, and BAY 11-7085, which inhibits IkappaB phosphorylation. Pneumococcal meningitis was associated with an increase in NF-kappaB activity, as determined by immunohistochemistry and Western blot analysis of rat brains 24 h after infection. Treatment with ALLN or BAY 11-7085 improved the clinical scores of infected rats, compared with those of untreated infected rats. This beneficial effect was parallelled by a significant reduction of the increase in intracranial pressure, blood-brain barrier permeability (as measured by the Evans blue extravasation technique), cerebrospinal fluid (CSF) pleocytosis, CSF interleukin 6 levels, and impairment of cerebrovascular CO(2) reactivity and autoregulation. Thus, pharmacologic interference with NF-kappaB activation might be a possible target for adjunctive therapy in bacterial meningitis. PMID- 11023467 TI - Helicobacter pylori vacA and cagA genotypes in Mexican adults and children. AB - Studies examining associations between Helicobacter pylori virulence markers and disease have concentrated on adults in developed countries. This study assessed adults and children in Mexico. Ninety patients were recruited, 56 adults (37 with active peptic ulceration and 19 with no ulcers) and 34 children (all with recurrent abdominal pain and no ulcers). H. pylori was cultured from gastric biopsy specimens, and vacA alleles and cagA were typed by use of polymerase chain reaction from multiple colony sweeps. Multiple vacA types were common in single biopsy isolates and were more frequent in adults with ulcers (95%) than in adults without ulcers (37%; P<.001) or in children (52%; P<.01). vacA s1b and cagA+ strains were more frequent in adults than in children. vacA s1 and cagA+ strains had similar frequencies in adults with and without ulcers. In conclusion, infection with multiple H. pylori strains, defined by different vacA genotypes, is common in Mexico. Such mixed infection is associated with ulcer disease. Strain populations infecting Mexican adults and children differ. PMID- 11023468 TI - Antigenic conservation of an immunodominant invariable region of the VlsE lipoprotein among European pathogenic genospecies of Borrelia burgdorferi SL. AB - Lyme disease is caused by genetically divergent spirochetes, including 3 pathogenic genospecies: Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii. Serodiagnosis is complicated by this genetic diversity. A synthetic peptide (C(6)), based on the 26-mer invariable region (IR(6)) of the variable surface antigen of B. burgdorferi (VlsE), was used as ELISA antigen, to test serum samples collected from mice experimentally infected with the 3 genospecies and from European patients with Lyme disease. Regardless of the infecting strains, mice produced a strong antibody response to C(6), which indicates that IR(6) is antigenically conserved among the pathogenic genospecies. Twenty of 23 patients with culture-confirmed erythema migrans had a detectable antibody response to C(6). A sensitivity of 95.2% was achieved, with serum samples collected from patients with well-defined acrodermatitis chronica atrophicans. Fourteen of 20 patients with symptoms of late Lyme disease also had a positive anti-IR(6) ELISA. Thus, it is possible that C(6) may be used to serodiagnose Lyme disease universally. PMID- 11023469 TI - T cell-monocyte contact enhances tumor necrosis factor-alpha production in response to Mycobacterium leprae. AB - Tumor necrosis factor (TNF)-alpha has been implicated as a key factor in inflammatory processes occurring in erythema nodosum leprosum (ENL). In the present study, the roles of soluble factors and contact-mediated interaction in the induction of enhanced TNF-alpha secretion in leprosy have been investigated. In vitro studies have demonstrated that Mycobacterium leprae per se is a poor stimulus for TNF-alpha production by purified monocytes obtained from normal subjects, although this could be enhanced by either exogenous interferon-gamma or cell contact with fixed activated T lymphocytes. Further investigations demonstrated that monocyte-T cell contact enhanced M. leprae-induced TNF-alpha production by peripheral blood mononuclear cells of ENL patients and was modulated by blocking antibodies to CD40L, CD69, and CD18. These results suggest that physical contact with T cells isolated from patients in a particular disease state (ENL) modulates monocyte function and may contribute to the secretion of proinflammatory cytokines described in ENL. PMID- 11023470 TI - Surfactant protein genetic marker alleles identify a subgroup of tuberculosis in a Mexican population. AB - Pulmonary surfactant and its components are essential for normal lung function and are involved in local host defense. Surfactant protein (SP)-A and SP-D bind to and modulate phagocytosis of Mycobacterium tuberculosis by macrophages. Frequency comparisons of SP marker alleles in tuberculosis patients and healthy control subjects (tuberculin-skin test positive or general population) were performed. Regression analyses of the tuberculosis and the tuberculin-skin test positive groups revealed, on the basis of odds ratios, tuberculosis susceptibility (DA11_C and GATA_3) and protective (AAGG_2) marker alleles. Similarly, between tuberculosis patients and general population control subjects, susceptibility 1A(3), 6A(4), and B1013_A and protective AAGG_1, and AAGG_7 marker alleles were observed. Moreover, interactions were seen between alleles 6A(2) and 1A(3) (P=.0064) and between 1A(3) and B1013_A (P=. 036). The findings indicate a possible involvement of SP alleles in tuberculosis pathogenesis. PMID- 11023471 TI - Growth inhibition of Candida by human oral epithelial cells. AB - Oropharyngeal candidiasis (OPC) caused by Candida albicans is a significant problem in human immunodeficiency virus (HIV)-infected persons. Recognizing the paucity of information on innate and/or adaptive mucosal host defenses against C. albicans, we recently reported that human and nonhuman primate and mouse vaginal epithelial cells inhibit the growth of C. albicans in vitro. In the present study, oral epithelial cells collected from saliva of healthy volunteers and a purified oral epithelial cell line were found to inhibit blastoconidia and/or hyphal growth of several Candida species. Cell contact was a strict requirement for the epithelial cell anti-Candida activity; neither saliva nor culture supernatants alone inhibited Candida growth, and addition of saliva to the coculture did not modulate the epithelial cell activity. Finally, epithelial cell anti-Candida activity was significantly lower in HIV-infected persons with OPC. Together, these results suggest that oral epithelial cells may play a role in innate resistance against OPC. PMID- 11023472 TI - Synthetic malaria peptide vaccine elicits high levels of antibodies in vaccinees of defined HLA genotypes. AB - A multiple antigen peptide (MAP) malaria vaccine containing minimal Plasmodium falciparum circumsporozoite protein repeat epitopes was assessed for safety and immunogenicity in volunteers of known class II genotypes. The MAP/alum/QS-21 vaccine formulation elicited high levels of parasite-specific antibodies in 10 of 12 volunteers expressing DQB1*0603, DRB1*0401, or DRB1*1101 class II molecules. In contrast, volunteers of other HLA genotypes were low responders or nonresponders. A second study of 7 volunteers confirmed the correlation of class II genotype and high responder phenotype. This is the first demonstration in humans that a peptide vaccine containing minimal T and B cell epitopes composed of only 5 amino acids (N, A, V, D, and P) can elicit antibody titers comparable to multiple exposures to irradiated P. falciparum-infected mosquitoes. Moreover, the high-responder phenotypes were predicted by analysis of peptide/HLA interactions in vitro, thus facilitating the rational design of epitope-based peptide vaccines for malaria, as well as for other pathogens. PMID- 11023473 TI - Interleukin-12 regulates the response to chemotherapy in experimental visceral Leishmaniasis. AB - In experimental visceral leishmaniasis, interleukin (IL)-12 initiates control over Leishmania donovani via Th1 cell activation, interferon (IFN)-gamma secretion, and granuloma formation. Because the leishmanicidal effect of conventional therapy, pentavalent antimony (Sb), also requires T cells and endogenous IFN-gamma, we tested IL-12 as a determinant of host responsiveness to chemotherapy. L. donovani-challenged IL-12p35 gene knockout (KO) mice permitted uncontrolled hepatic infection and failed to respond to Sb. In contrast, 96% of liver parasites in KO mice were killed by amphotericin B, which acts independently of immune responses. Exogenous IL-12 combined with Sb was tested in normal mice: low-dose Sb was converted from weakly to strongly leishmanicidal, and a no-effect Sb dose was converted to approximately 100% leishmanistatic. IL 12 plus Sb synergism in normal mice was IFN-gamma dependent; however, IL-12 also increased responsiveness to Sb in IFN-gamma KO mice. Thus, IL-12 regulates host IFN-gamma-dependent and -independent responses that permit and/or enhance the leishmanicidal activity of Sb. PMID- 11023474 TI - Prospective reevaluation of risk factors in mother-to-child transmission of hepatitis C virus: high virus load, vaginal delivery, and negative anti-NS4 antibody. AB - Of 21,791 pregnant women screened in Tottori Prefecture, Japan, 127 (0.58%) were positive for anti-hepatitis C virus (HCV) antibody and 84 (0.39%) were positive for HCV RNA. Of 84 children followed up for at least 6 months, 7 (8%) were infected. All of them were born to 26 mothers with a high virus load (HVL; >/=2.5x106 RNA copies/mL [27%]), compared with 0 of 58 children born to non-HVL mothers (P<.001). Because all the infected children were vaginally delivered, the infection rate among 16 vaginally delivered children born to the HVL mothers was as high as 44%. The prevalence of anti-NS4 antibody in the mothers with an infectious HVL was significantly lower than that in the mothers with a noninfectious HVL (P=.048). Analysis of our results suggests that maternal HVL, vaginal delivery, and negative anti-NS4 antibody are significant risk factors for the mother-to-child transmission of HCV. PMID- 11023475 TI - The amino acid sequence of the PKR-eIF2alpha phosphorylation homology domain of hepatitis C virus envelope 2 protein and response to interferon-alpha. AB - A region of the hepatitis C virus (HCV) envelope 2 protein, the protein kinase, PKR and early initiation factor 2alpha phosphorylation homology domain (PePHD), may be important in interferon (IFN)-alpha resistance. The PePHD was amplified by polymerase chain reaction and sequenced, and the amino acid sequence derived from pretreatment serum of 14 genotype 3-infected patients with a range of responses to IFN-alpha therapy. Only 1 patient had a PePHD variant. IFN-resistant PePHD variants present at low titers in pretreatment serum should be selected by therapy; therefore, the PePHD amino acid sequence was also obtained from serum collected during or after treatment in 5 patients with breakthrough or relapse of HCV RNA positivity. No difference was found between the pre- and posttreatment PePHD sequences. Thus, it appears that pretreatment sequencing of the PePHD would not enable clinicians to predict the treatment response. There was no evidence that IFN therapy exerts selection pressure in this region. PMID- 11023476 TI - Human caliciviruses are a significant pathogen of acute sporadic diarrhea in children of Santiago, Chile. AB - Human caliciviruses (HuCVs) are increasingly recognized as common pathogens that cause acute sporadic diarrhea in children; however, regional antigenic and genetic diversity complicate detection techniques. Stool samples from children seeking medical attention in 2 outpatient clinics, a large emergency department, and 2 hospital wards were evaluated for HuCVs by reverse transcription-polymerase chain reaction, using primers based on a conserved sequence of the polymerase region of a previously sequenced Chilean strain. HuCVs were detected in 53 (8%) of 684 children 1 month to 5 years of age (mean, 13 months). Detection occurred year-round without a clear seasonal peak, and detection frequency declined from 16% in 1997 to 2% in 1999. The decline may have been due to a change in virus genotype. HuCVs are a significant pathogen of acute sporadic diarrhea in Chilean children, and continuous characterization of genetic diversity will be crucial for appropriate detection. PMID- 11023477 TI - A randomized, placebo-controlled trial of granulocyte-macrophage colony stimulating factor and nucleoside analogue therapy in AIDS. AB - Preliminary preclinical and clinical data suggest that granulocyte-macrophage colony-stimulating factor (GM-CSF) may decrease viral replication. Therefore, 105 individuals with AIDS who were receiving nucleoside analogue therapy were enrolled in a placebo-controlled, double-blind study and were randomized to receive either 125 microgram/m(2) of yeast-derived, GM-CSF (sargramostim) or placebo subcutaneously twice weekly for 6 months. Subjects were evaluated for toxicity and disease progression. A significant decrease in mean virus load (VL) was observed for the GM-CSF treatment group at 6 months (-0.07 log(10) vs. -0.60 log(10); P=.02). More subjects achieved human immunodeficiency virus (HIV)-RNA levels <500 copies/mL at >/=2 evaluations (2% on placebo vs. 11% on GM-CSF; P=.04). Genotypic analysis of 46 subjects demonstrated a lower frequency of zidovudine-resistant mutations among those receiving GM-CSF (80% vs. 50%; P=.04). No difference was observed in the incidence of opportunistic infections (OIs) through 6 months or survival, despite a higher risk for OI among GM-CSF recipients. GM-CSF reduced VL and limited the evolution of zidovudine-resistant genotypes, potentially providing adjunctive therapy in HIV disease. PMID- 11023478 TI - Ex vivo and in vitro effect of human immunodeficiency virus protease inhibitors on neutrophil apoptosis. AB - Polymorphonuclear leukocytes (PMNL) from human immunodeficiency virus (HIV) infected patients exhibit accelerated apoptosis and impaired functional activity. HIV protease inhibitor-based therapy produces improvements in both acquired and innate immune responses. Ex vivo and in vitro effects of HIV protease inhibitors on apoptosis and chemotaxis of PMNL were evaluated. After therapy, there was a rapid and significant decrease of PMNL apoptosis, which correlated with increased chemotactic function. These findings were found both in patients with immunological and virological response and in control subjects who showed an increase in CD4(+) T cell counts but no concomitant decline in HIV load. After in vitro treatment with ritonavir or indinavir, apoptosis of both HIV-infected and uninfected PMNL markedly decreased and correlated with significant enhancement of chemotaxis. These results suggest that HIV protease inhibitors may improve the PMNL function by reducing the apoptosis rate and that this effect may, at least in part, be independent of their antiviral activity. PMID- 11023479 TI - Intestinal infection due to enteroaggregative Escherichia coli among human immunodeficiency virus-infected persons. AB - To investigate the pathogenic role of enteroaggregative Escherichia coli (EAggEC) among human immunodeficiency virus-infected persons, 111 outpatients with and 68 without diarrhea were evaluated. Examination of stool samples included the HeLa cell adherence assay and an EAggEC polymerase chain reaction (PCR) assay using primers complementary for the plasmid locus CVD432. The pCVD432 genotype, adherence phenotype, and patient characteristics were correlated with occurrence of diarrhea by multivariate analyses. EAggEC PCR and adherence assays were positive in 7 (6%) and 24 (22%) patients with diarrhea and in 1 (1%) and 21 (31%) asymptomatic control patients, respectively. Clinical manifestations associated with EAggEC PCR-positive isolates were nonspecific; EAggEC infections were independent of CD4 lymphocyte counts. Of the pCVD432 genotype, 5 (71%) of 7 were resistant to cotrimoxazole and ampicillin, and 1 strain was resistant to ciprofloxacin. Overall, pCVD432 PCR-positive E. coli was the most prevalent intestinal organism associated with diarrhea. The adherence assay results did not correlate with diarrhea. PMID- 11023480 TI - Polymorphisms in candidate genes and risk of scarring trachoma in a Chlamydia trachomatis--endemic population. AB - Genes involved in regulating antimicrobial immunity and inflammation may modulate the risk of tissue scarring and fibrosis in chlamydial diseases such as trachoma. By use of a large case-control study of scarring trachoma in The Gambia, the importance of single-nucleotide polymorphisms in several candidate genes was investigated. Overall, no significant differences were found between patients and control subjects in genotype frequencies for polymorphisms in cytokine promoters interleukin (IL)-10 (positions -1082, -819, -592), IL-4 (-590), or tumor necrosis factor-alpha (-376) or for codon 57 of the mannose-binding protein. Among the ethnic groups in the study, Mandinkas had the highest frequency of the IL-10 1082G allele (0.36). Within this ethnic group, the IL-10-1082G homozygote genotype was significantly more common among case patients than control subjects (odds ratio, 5.10; 95% confidence interval, 1.24-24.2; P=.009). This single association is consistent with data indicating that the IL-10-1082G allele is associated with higher levels of IL-10 transcription and that Th2-type immune responses are associated with risk of chlamydial disease. PMID- 11023481 TI - Prevention of Haemophilus influenzae type b colonization by vaccination: correlation with serum anti-capsular IgG concentration. AB - Concentrations of serum anti-Haemophilus influenzae type b (anti-Hib) capsular polysaccharide (CPS) >/=0.15 and >/=1.0 microgram/mL are widely used as surrogates for protection against invasive Hib disease. However, the relationship between serum anti-Hib CPS following immunization and protection against colonization is not known, making it difficult to evaluate new Hib vaccines or combination vaccines. In the Dominican Republic, nasopharyngeal swabs were collected from 546 9-month-old infants who had received Hib conjugate vaccine at ages 2, 4, and 6 months and from 600 unvaccinated infants of the same age. The prevalence of Hib colonization was lower among vaccinated infants than among unvaccinated infants (0.9% vs. 2.3%). Among vaccinated infants, protection against colonization was significantly correlated with anti-Hib CPS concentrations >/=5 microgram/mL 1 month following the third dose of vaccine. These results suggest that the concentration of serum anti-Hib CPS needed for protection against colonization is greater than that needed for protection for invasive disease. PMID- 11023482 TI - An interleukin-1 genotype is associated with fatal outcome of meningococcal disease. AB - To determine whether known variants of the interleukin-1 (IL-1) and tumor necrosis factor (TNF) gene families are associated with severe manifestations of meningococcal disease, 276 white patients 4-70 years of age (median, 17 years) were genotyped. All patients had microbiologically proven Neisseria meningitidis infection; 39 died and 237 survived. A significant association (P<.001) was found between fatal outcome and genotype at IL1B (nucleotide position -511). Homozygous individuals, both for the common (1/1) and the rare (2/2) alleles, had increased odds ratios (ORs) for death, compared with heterozygous individuals (1/2): ORs (95% confidence intervals [CIs]) were 3.39 (1.39-8.29) and 7.35 (2.51-21.45), respectively. The mortality rates according to genotype at IL1B (-511) were 18.0% (1/1), 6.1% (1/2), and 32.3% (2/2), compared with 14.2% overall. The composite genotype, consisting of heterozygosity of IL1B (-511) together with homozygosity of the common allele of the IL-1 receptor antagonist gene (IL1RN) at +2018, was significantly associated with survival (P=.018; OR, 7.78 [95% CI, 1. 05-59.05]). There was no association between TNF genotype and fatal outcome. These data suggest that IL-1 genotype influences the severity of meningococcal disease. PMID- 11023483 TI - Effects of antibiotic class on the macrophage inflammatory response to Streptococcus pneumoniae. AB - Antibiotic choice can alter host inflammation during invasive bacterial infections. Previous studies of gram-negative organisms concluded that antibiotic mediated release of bacterial cell wall components amplifies inflammation. Less has been reported about antibiotic effect on gram-positive organisms. This study explored the hypothesis that Streptococcus pneumoniae would induce greater macrophage inflammatory mediator production when killed with cell wall active antibiotics rather than protein synthesis inhibitors. Stimulation of RAW 264.7 murine macrophages with pneumococci and oxacillin led to significantly higher inducible nitric oxide synthase (iNOS) and tumor necrosis factor (TNF) accumulation than did the same concentrations of pneumococci and clindamycin. Neither antibiotic alone or in combination with lipopolysaccharide acted directly on macrophages to modify the immune response. Endotoxin contamination did not confound the results, as preincubation with polymyxin B did not change iNOS or TNF protein levels. Thus, the antimicrobial mechanism of action affects macrophage inflammatory mediator production after stimulation with pneumococci. PMID- 11023484 TI - Identification of loci essential for the growth of Helicobacter pylori under acidic conditions. AB - To persist in the hostile acidic environment of the stomach, Helicobacter pylori must survive acid shock and grow at acidic pH. Of a library of 1250 random mutants screened for isolates unable to grow at low pH, 10 mutants were detected that were unable to grow at pH 4.8. However, all 10 mutants were resistant to acid shock. Four mutants had an insertion in genes of unknown function. One mutant was affected in lepA, an orthologue of a membrane GTPase. Three mutants were disrupted in loci involved in the transport of H(+) ions or other cations (FRaseI, czcA, and aldo-keto reductase). Two mutants were affected in loci that contribute to acid resistance in other microorganisms (uvrA and atpF'). Thus, at least 10 loci not related to urease are essential for the growth of H. pylori under acidic conditions and should be critical for lifelong infection by this pathogen. PMID- 11023485 TI - Plasma interleukin-10:Tumor necrosis factor (TNF)-alpha ratio is associated with TNF promoter variants and predicts malarial complications. AB - In individuals with severe malarial anemia, plasma levels of tumor necrosis factor (TNF)-alpha tend to exceed those of interleukin (IL)-10. In this study, IL 10:TNF plasma level ratios <1 were found to be a risk factor for both cerebral malaria and severe anemia (P=.009), whereas higher IL-10:TNF ratios were observed more frequently in hyperparasitemic individuals. When considering allelic variants of the TNF promoter in children with severe malaria, carriers of the wild type more frequently had an IL-10:TNF ratio >1 (P=.008). In contrast, individuals with a mutation at position -238 of the TNF promoter (TNF(-238A) and TNF(-376A/-238A)) consistently had lower IL-10 than TNF plasma levels (IL-10:TNF ratio <1; P=.003). Our results show that, in children with severe malaria, TNF promoter variants influence the balance of IL-10:TNF in the plasma, which, in turn, affects the outcome in terms of clinical complications. PMID- 11023486 TI - Reply PMID- 11023487 TI - Human herpesvirus 8 in Brazilian Amerindians: does vertical transmission play a role? PMID- 11023488 TI - Are only antibody levels involved in the protection against pertussis in acellular pertussis vaccine recipients? PMID- 11023489 TI - Reply PMID- 11023490 TI - Chlamydia pneumoniae viability in atherosclerotic tissue: true or false? PMID- 11023491 TI - Reply PMID- 11023492 TI - CCR2-64I polymorphism, syncytium-inducing human immunodeficiency virus strains, and disease progression. PMID- 11023493 TI - PU.1 inhibits GATA-1 function and erythroid differentiation by blocking GATA-1 DNA binding. AB - The lineage-specific transcription factors GATA-1 and PU.1 can physically interact to inhibit each other's function, but the mechanism of repression of GATA-1 function by PU.1 has not been elucidated. Both the N terminus and the C terminus of PU.1 can physically interact with the C-terminal zinc finger of GATA 1. It is demonstrated that the PU.1 N terminus, but not the C terminus, is required for inhibiting GATA-1 function. Induced overexpression of PU.1 in K562 erythroleukemia cells blocks hemin-induced erythroid differentiation. In this system, PU.1 does not affect the expression of GATA-1 messenger RNA, protein, or nuclear localization. However, GATA-1 DNA binding decreases dramatically. By means of electrophoretic mobility shift assays with purified proteins, it is demonstrated that the N-terminal 70 amino acids of PU.1 can specifically block GATA-1 DNA binding. In addition, PU.1 had a similar effect in the G1ER cell line, in which the GATA-1 null erythroid cell line G1E has been transduced with a GATA 1-estrogen receptor fusion gene, which is directly dependent on induction of the GATA-1 fusion protein to effect erythroid maturation. Consistent with in vitro binding assays, overexpression of PU.1 blocked DNA binding of the GATA-1 fusion protein as well as GATA-1-mediated erythroid differentiation of these G1ER cells. These results demonstrate a novel mechanism by which function of a lineage specific transcription factor is inhibited by another lineage-restricted factor through direct protein-protein interactions. These findings contribute to understanding how protein-protein interactions participate in hematopoietic differentiation and leukemogenesis. (Blood. 2000;96:2641-2648) PMID- 11023494 TI - Up-regulation of HIV coreceptors CXCR4 and CCR5 on CD4(+) T cells during human endotoxemia and after stimulation with (myco)bacterial antigens: the role of cytokines. AB - Concurrent infections in patients with human immunodeficiency virus (HIV) infection stimulate HIV replication. Chemokine receptors CXCR4 and CCR5 can act as HIV coreceptors. The authors hypothesized that concurrent infection increases the HIV load through up-regulation of CXCR4 and CCR5. Using experimental endotoxemia as a model of infection, changes in HIV coreceptor expression were assessed in 8 subjects injected with lipopolysaccharide (LPS, 4 ng/kg). The expression of CXCR4 and CCR5 on CD4(+) T cells was increased 2- to 4-fold, 4 to 6 hours after LPS injection. In whole blood in vitro, LPS induced a time- and dose dependent increase in the expression of CXCR4 and CCR5 on CD4(+) T cells. Similar changes were observed after stimulation with cell wall components of Mycobacterium tuberculosis (lipoarabinnomannan) or Staphylococcus aureus (lipoteichoic acid), or with staphylococcal enterotoxin B. LPS increased viral infectivity of CD4-enriched peripheral blood mononuclear cells (PBMCs) with a T tropic HIV strain. In contrast, M-tropic virus infectivity was reduced, possibly because of elevated levels of the CCR5 ligand cytokines RANTES and MIP-1beta. LPS stimulated up-regulation of CXCR4 and CCR5 in vitro was inhibited by anti-TNF and anti-IFN gamma. Incubation with recombinant TNF or IFN gamma mimicked the LPS effect. Anti-interleukin 10 (anti-IL-10) reduced CCR5 expression, without influencing CXCR4. In accordance, rIL-10 induced up-regulation of CCR5, but not of CXCR4. Intercurrent infections during HIV infection may up-regulate CXCR4 and CCR5 on CD4(+) T cells, at least in part via the action of cytokines. Such infections may favor selectivity of HIV for CD4(+) T cells expressing CXCR4. (Blood. 2000;96:2649-2654) PMID- 11023495 TI - Blood-derived nurse-like cells protect chronic lymphocytic leukemia B cells from spontaneous apoptosis through stromal cell-derived factor-1. AB - A subset of blood cells from patients with B-cell chronic lymphocytic leukemia (CLL) spontaneously differentiates in vitro into large, round, or fibroblast-like adherent cells that display stromal cell markers, namely vimentin and STRO-1. These cells also express stromal cell-derived factor-1 (SDF-1), a CXC chemokine that ordinarily is secreted by marrow stromal cells. Leukemia B cells attach to these blood-derived adherent cells, down-modulate their receptors for SDF-1 (CXCR4), and are protected from undergoing spontaneous apoptosis in vitro. Neutralizing antibodies to SDF-1 inhibit this effect. Moreover, the rapid deterioration in the survival of CLL B cells, when separated from such cells, is mitigated by exogenous SDF-1. This chemokine also results in the rapid down modulation of CXCR4 and activation of p44/42 mitogen-activated protein-kinase (ERK 1/2) by CLL B cells in vitro. It is concluded that the blood of patients with CLL contains cells that can differentiate into adherent nurse-like cells that protect leukemia cells from undergoing spontaneous apoptosis through an SDF 1-dependent mechanism. In addition to its recently recognized role in CLL B-cell migration, SDF-1-mediated CLL B-cell activation has to be considered a new mechanism involved in the microenvironmental regulation of CLL B-cell survival. (Blood. 2000;96:2655-2663) PMID- 11023496 TI - Antimicrobial proteins and peptides of blood: templates for novel antimicrobial agents. AB - The innate immune system provides rapid and effective host defense against microbial invasion in a manner that is independent of prior exposure to a given pathogen. It has long been appreciated that the blood contains important elements that mediate rapid responses to infection. Thus, anatomic compartments with ample blood supply are less frequently infected and recover more readily once infected, whereas regions with poor perfusion are prone to severe infection and may require surgical debridement. Blood-borne innate immune mediators are either carried in circulating blood cells (ie, leukocytes and platelets) or in plasma after release from blood cells or on secretion by the liver. (Blood. 2000;96:2664-2672) PMID- 11023497 TI - Neutrophil gelatinase B potentiates interleukin-8 tenfold by aminoterminal processing, whereas it degrades CTAP-III, PF-4, and GRO-alpha and leaves RANTES and MCP-2 intact. AB - Chemokines are mediators in inflammatory and autoimmune disorders. Aminoterminal truncation of chemokines results in altered specific activities and receptor recognition patterns. Truncated forms of the CXC chemokine interleukin (IL)-8 are more active than full-length IL-8 (1-77), provided the Glu-Leu-Arg (ELR) motif remains intact. Here, a positive feedback loop is demonstrated between gelatinase B, a major secreted matrix metalloproteinase (MMP-9) from neutrophils, and IL-8, the prototype chemokine active on neutrophils. Natural human neutrophil progelatinase B was purified to homogeneity and activated by stromelysin-1. Gelatinase B truncated IL-8(1-77) into IL-8(7-77), resulting in a 10- to 27-fold higher potency in neutrophil activation, as measured by the increase in intracellular Ca(++) concentration, secretion of gelatinase B, and neutrophil chemotaxis. This potentiation correlated with enhanced binding to neutrophils and increased signaling through CXC chemokine receptor-1 (CXCR1), but it was significantly less pronounced on a CXCR2-expressing cell line. Three other CXC chemokines-connective tissue-activating peptide-III (CTAP-III), platelet factor-4 (PF-4), and GRO-alpha-were degraded by gelatinase B. In contrast, the CC chemokines RANTES and monocyte chemotactic protein-2 (MCP-2) were not digested by this enzyme. The observation of differing effects of neutrophil gelatinase B on the proteolysis of IL-8 versus other CXC chemokines and on CXC receptor usage by processed IL-8 yielded insights into the relative activities of chemokines. This led to a better understanding of regulator (IL-8) and effector molecules (gelatinase B) of neutrophils and of mechanisms underlying leukocytosis, shock syndromes, and stem cell mobilization by IL-8. (Blood. 2000;96:2673-2681) PMID- 11023498 TI - Presentation of chemokine SDF-1 alpha by fibronectin mediates directed migration of T cells. AB - The role of chemokine-matrix interactions in integrin-dependent T-cell migration was examined to address the critical question of how chemokines provide directional information. The chemokine SDF-1 alpha binds fibronectin (Fn) with a low nanomolar K(d) (equilibrium dissociation constant). SDF-1 alpha presented by Fn induced directed migration. Spatial concentration gradients of chemokine were not required to maintain directed migration. Fn-presented chemokine induced the polarization of cells, including the redistribution of the SDF-1 alpha receptor, to the basal surface and leading edge of the cell. A new model for directed migration is proposed in which the co-presentation of an adhesive matrix and chemokine provides the necessary positional information independent of a soluble spatial gradient. (Blood. 2000;96:2682-2690) PMID- 11023499 TI - Clinical importance of minimal residual disease in childhood acute lymphoblastic leukemia. AB - By using rapid flow cytometric techniques capable of detecting one leukemic cell in 10(4) normal cells, we prospectively studied minimal residual disease (MRD) in 195 children with newly diagnosed acute lymphoblastic leukemia (ALL) in clinical remission. Bone marrow aspirates (n = 629) were collected at the end of remission induction therapy and at 3 intervals thereafter. Detectable MRD (ie, > or = 0.01% leukemic mononuclear cells) at each time point was associated with a higher relapse rate (P < .001); patients with high levels of MRD at the end of the induction phase (> or = 1%) or at week 14 of continuation therapy (> or = 0.1%) had a particularly poor outcome. The predictive strength of MRD remained significant even after adjusting for adverse presenting features, excluding patients at very high or very low risk of relapse from the analysis, and considering levels of peripheral blood lymphoblasts at day 7 and day 10 of induction therapy. The incidence of relapse among patients with MRD at the end of the induction phase was 68% +/- 16% (SE) if they remained with MRD through week 14 of continuation therapy, compared with 7% +/- 7% if MRD became undetectable (P = .035). The persistence of MRD until week 32 was highly predictive of relapse (all 4 MRD(+) patients relapsed vs 2 of the 8 who converted to undetectable MRD status; P = .021). Sequential monitoring of MRD by the method described here provides highly significant, independent prognostic information in children with ALL. Recent improvements in this flow cytometric assay have made it applicable to more than 90% of all new patients. (Blood. 2000;96:2691-2696) PMID- 11023500 TI - Biochemical markers of bone turnover following high-dose chemotherapy and autografting in multiple myeloma. AB - The effect of high-dose chemotherapy and autografting on bone turnover in myeloma is not known. A study of 32 myeloma patients undergoing blood or marrow transplant (BMT), conditioned with high-dose melphalan, was done. Bone resorption was assessed by urinary free pyridinoline (fPyr) and deoxypyridinoline (fDPyr), expressed as a ratio of the urinary creatinine concentration. Bone formation was assessed by serum concentration of procollagen 1 extension peptide (P1CP) and bone-specific alkaline phosphatase (BSAP). Eighteen cases had normal fPyr and fDPyr at transplant, and in all but one of these cases the level remained normal throughout subsequent follow-up. In contrast, in 14 cases urinary fPyr and fDPyr levels were increased at transplant. In these cases, both fPyr and fDPyr fell to normal levels over the next few months (P = .0009 and.0019, respectively). fPyr and fDPyr levels at transplant and their trends post-BMT were unrelated to the use of pre-BMT or post-BMT bisphosphonate or post-BMT interferon. Nine cases had elevated P1CP or BSAP at transplant, which rapidly normalized. In most patients there was an increase in P1CP and/or BSAP several months post-transplant. In conclusion, increased osteoclast activity may be present even in apparent plateau phase of myeloma. High-dose chemotherapy with autografting may normalize abnormal bone resorption, although the effect may take several weeks to emerge and may be paralleled by increased osteoblast activity. The findings provide biochemical evidence that autografting may help normalize the abnormal bone turnover characteristic of myeloma. (Blood. 2000;96:2697-2702) PMID- 11023501 TI - Analysis of engraftment, graft-versus-host disease, and immune recovery following unrelated donor cord blood transplantation. AB - Unrelated cord blood (UCB) is being used as a source of alternative hematopoietic stem cells for transplantation with increasing frequency. From November 1994 to February 1999, 30 UCB transplant procedures were performed for both malignant and nonmalignant diseases in 27 children, aged 0.4 to 17.1 years. Patients received either HLA-matched (n = 3) or 1- or 2-antigen-mismatched (n = 27) UCB following 1 of 2 standardized preparative and graft-versus-host disease regimens (hyperfractionated total body irradiation, cyclophosphamide, and antithymocyte globulin [ATG] with cyclosporine A and methotrexate; or busulfan, melphalan, and ATG with cyclosporine A and prednisone). The median time to neutrophil and platelet engraftment was 27 days (12-60 days) and 75 days (33-158 days) posttransplantation, respectively. No correlation was noted between neutrophil and platelet engraftment and nucleated cells per kilogram, CD34(+) cells per kilogram infused, or cytomegalovirus status of recipient. The cumulative probability of acute grade 2 or greater graft-versus-host disease (GVHD) was 37.2%, and of grade 3 or greater GVHD was 8.8%. No patients developed chronic GVHD. CD4, CD19, and natural killer cell recovery was achieved at a median of 12, 6, and 2 months, respectively. CD8 recovery was delayed at a median of 9 months. Normal mitogen response was achieved at 6 to 9 months. The probability of survival, disease-free survival, and event-free survival at 1 year was 52.3% (34.1%-70.5%), 54.7% (34.5%-74.9 %) and 49.6% (29.9%-69.4%), respectively. This series of 30 UCB transplants suggests that although CD8 cell recovery is delayed, the pattern of immune reconstitution with UCB is similar to that reported for other stem cell sources. (Blood. 2000;96:2703-2711) PMID- 11023502 TI - Durability of responses following donor lymphocyte infusions for patients who relapse after allogeneic stem cell transplantation for chronic myeloid leukemia. AB - An analysis was performed of the response to treatment with donor lymphocyte infusions (DLI) and the survival in 66 consecutive patients who relapsed after primary treatment by allogeneic stem cell transplantation for BCR-ABL-positive chronic myeloid leukemia. The transplant donor was an HLA-identical sibling (n = 35) or a "matched" unrelated volunteer (n = 31). Fifty-seven patients were transplanted in chronic phase, eight in accelerated phase, and one in second chronic phase. The recognition of relapse was based on precise molecular, cytogenetic, or hematologic criteria. The median interval from transplant to relapse was 12 months (range 3-85). The median interval from relapse to initiation of DLI was 9.4 months (range 1-70). Patients received DLI from their original transplant donors on a bulk-dose (n = 34) or on an escalating-dose (n = 32) regimen. Patients were monitored serially by hematologic, cytogenetic, and molecular criteria. Molecular remission was defined by the finding of negative results by nested primer reverse transcriptase polymerase chain reaction (RT-PCR) for BCR-ABL transcripts on two consecutive occasions, subject to satisfactory controls. Forty-four patients (67%) achieved molecular remission. Patients who had relapsed to advanced phase disease and patients with short intervals between transplant and relapse had significantly lower probabilities of achieving molecular remission. Of the 44 patients who achieved molecular remission, 4 reverted to a PCR-positive status at 15, 18, 37, and 87 weeks after remission. The probability of survival for patients who achieved molecular remission was significantly better than for those who failed to do so (95% versus 53% at 3 years post-DLI, P = .0001). We conclude that the majority of molecular remissions after DLI are durable, and thus the majority of responding patients may prove to have been cured. (Blood. 2000;96:2712-2716) PMID- 11023503 TI - Cell dose and speed of engraftment in placental/umbilical cord blood transplantation: graft progenitor cell content is a better predictor than nucleated cell quantity. AB - There is evidence that the total cellular content of placental cord blood (PCB) grafts is related to the speed of engraftment, though the total nucleated cell (TNC) dose is not a precise predictor of the time of neutrophil or platelet engraftment. It is important to understand the reasons for the quantitative association and to improve the criteria for selecting PCB grafts by using indices more precisely predictive of engraftment. The posttransplant course of 204 patients who received grafts evaluated for hematopoietic colony-forming cell (CFC) content among 562 patients reported previously were analyzed using univariate and multivariate life-table techniques to determine whether CFC doses predicted hematopoietic engraftment speed and risk for transplant-related events more accurately than the TNC dose. Actuarial times to neutrophil and platelet engraftment were shown to correlate with the cell dose, whether estimated as TNC or CFC per kilogram of recipient's weight. CFC association with the day of recovery of 500 neutrophils/microL, measured as the coefficient of correlation, was stronger than that of the TNC (R = -0.46 and -0.413, respectively). In multivariate tests of speed of platelet and neutrophil engraftment and of probability of posttransplantation events, the inclusion of CFC in the model displaced the significance of the high relative risks associated with TNC. The CFC content of PCB units is associated more rigorously with the major covariates of posttransplantation survival than is the TNC and is, therefore, a better index of the hematopoietic content of PCB grafts. (Blood. 2000;96:2717-2722) PMID- 11023504 TI - Cladribine with prednisone versus chlorambucil with prednisone as first-line therapy in chronic lymphocytic leukemia: report of a prospective, randomized, multicenter trial. AB - The efficacy and toxicity of cladribine (2-CdA) + prednisone (P) versus chlorambucil (Chl) + P were compared in previously untreated patients with progressive or symptomatic chronic lymphocytic leukemia (CLL) in a randomized, multicenter prospective trial. Eligible patients were assigned to either 2-CdA 0.12 mg/kg per day in 2-hour infusions and P 30 mg/m(2) per day for 5 consecutive days or Chl 12 mg/m(2) per day and P 30 mg/m(2) per day for 7 consecutive days. Three courses were administered at 28-day intervals or longer if myelosuppression developed. The therapy was finished if complete response (CR) was achieved. Of 229 available patients 126 received 2-CdA+P and 103 received Chl+P as a first line treatment. CR and overall response rates were significantly higher in the patients treated with 2-CdA+P (47% and 87%, respectively) than in the patients treated with Chl+P (12% and 57%, respectively) (P = .001). Progression-free survival was significantly longer in the 2-CdA-treated group (P = .01), but event free survival was not statistically different. Thirteen percent of patients were refractory to 2-CdA+P and 43% to Chl+P (P = .001). Drug-induced neutropenia was more frequently observed during 2-CdA+P (23%) than Chl+P therapy (11%) (P = .02), but thrombocytopenia occurred with similar frequency in both groups (36% and 27%, respectively). Infections were seen more frequently in the 2-CdA+P-treated group (56%) than in the Chl+P-treated group (40%; P = .02). Death rates have so far been similar in patients treated with 2-CdA (20%) and with Chl (17%). The probability of overall survival calculated from Kaplan-Meier curves at 24 months was also similar for both groups (78% and 82%, respectively). (Blood. 2000;96:2723-2729) PMID- 11023505 TI - Changes in acquired immunodeficiency syndrome-related lymphoma since the introduction of highly active antiretroviral therapy. AB - Clinical data on 7840 HIV-positive patients, representing 43 745 patient-years of follow-up, has been collected. All patients with ARL since 1986 (n = 150) were assessed at presentation for prognostic factors and outcomes recorded. Comparisons are made between cases in the pre-HAART era (1988-1995), and the HAART era (1996-1999). Statistical models are used to calculate the incidence of ARL and factors predicting its development. The incidence of ARL has not changed over time (3 to 7 of 1000 patients per year, P = .933), but contributes to a greater percentage of first AIDS-defining illnesses (ADI) in the HAART era (P < or = .0001). Older age, nadir CD4 count, and no prior HAART use, predict the development of ARL. There has been no change in stage at presentation, presence of B symptoms, performance status, or marrow involvement between the 2 time cohorts or between patients with or without prior HAART exposure. Similarly, there is no difference in survival duration between the pre-HAART and HAART era (log rank P = .15) or specifically in patients treated with HAART before ARL diagnosis (log rank P = .12). The use of HAART has not yet been shown to influence the incidence or survival of ARL. However, because nadir CD4 count and use of HAART are independent predictors of ARL development, this may translate into a future fall in new cases. (Blood. 2000;96:2730-2734) PMID- 11023506 TI - Protease inhibitors stimulate hematopoiesis and decrease apoptosis and ICE expression in CD34(+) cells. AB - Highly active retroviral therapy has been associated with a decline in the frequency of cytopenia in patients with human immunodeficiency virus (HIV) infection. This may result from lower hematologic toxicity of newer antiviral drugs and their increased efficacy against HIV-1. Protease inhibitors, in addition to their effects on HIV replication, appear to affect various cellular functions. Recently, it was reported that ritonavir inhibited caspase-1 expression in normal CD4(+) cells. It was hypothesized that protease inhibitors may improve hematopoietic function owing to their direct effects on the bone marrow progenitor cells. When ritonavir was added to methylcellulose cultures of bone marrow cells from HIV-infected patients and normal controls, colony formation increased 2.4-fold (n = 5) in control cultures and 4-fold (n = 5) in cultures of cells from HIV-infected patients. In the presence of ritonavir, cultures of CD34(+) cells showed markedly decreased apoptosis in comparison with untreated cultures (45% decrease in apoptotic cell number; n = 6). A synthetic inhibitor of caspase 1 (Ac-Tyr-Val-Ala-Asp-aldehyde [single-letter amino acid codes]), which inhibits activation of several caspases including CPP32 and interleukin 1beta-converting enzyme (ICE or caspase 1), also decreased the rate of apoptosis and enhanced colony formation by progenitor cells derived from HIV infected patients (3-fold; n = 5). In ritonavir-treated samples derived from HIV infected individuals, the number of cells expressing ICE also decreased. In conclusion, HIV protease inhibitors may, by blocking the caspase-dependent apoptotic pathway, overcome inhibition of hematopoiesis seen in patients with HIV infection, an effect unrelated to their antiviral activity. (Blood. 2000;96:2735 2739) PMID- 11023507 TI - Expression of the collagen receptor glycoprotein VI during megakaryocyte differentiation. AB - This study examined the expression of the platelet collagen receptor glycoprotein VI (GPVI) in megakaryocyte cell lines and primary megakaryocytes by reverse transcriptase-polymerase chain reaction and by flow cytometry and ligand blotting using the snake venom toxin convulxin. Expression of GPVI is increased in the megakaryoblastic cell lines HEL and CMK on differentiation with the phorbol ester phorbol 12-myristate 13-acetate (PMA), along with the Fc receptor gamma-chain (FcR gamma-chain). The increase in GPVI expression is associated with marked potentiation of tyrosine phosphorylation and Ca(++) elevation in response to convulxin. Syk, linker for activated T cells, and phospholipase C gamma 2 (PLC gamma 2) are among the proteins tyrosine phosphorylated on convulxin stimulation in PMA-differentiated HEL cells. Studies on primary murine megakaryocytes grown in vitro confirmed that GPVI is up-regulated in parallel with functional activation, assessed by measurement of [Ca(++)](i), during differentiation. The results demonstrate that expression of GPVI is up-regulated along with the FcR gamma-chain during differentiation of megakaryocytes. (Blood. 2000;96:2740-2745) PMID- 11023509 TI - Manipulating the onset of cell cycle withdrawal in differentiated erythroid cells with cyclin-dependent kinases and inhibitors. AB - Terminal differentiation of erythroid cells results in terminal cell divisions followed by irreversible cell cycle withdrawal of hemoglobinized cells. The mechanisms leading to cell cycle withdrawal were assessed in stable transfectants of murine erythroleukemia cells, in which the activities of cyclin-dependent kinases (CDKs) and CDK inhibitors (CDKIs) could be tightly regulated during differentiation. Cell cycle withdrawal of differentiating cells is mediated by induction of several CDKIs, thereby leading to inhibition of CDK2 and CDK4. Manipulation of CDK activity in differentiating cells demonstrates that the onset of cell cycle withdrawal can be either greatly accelerated or greatly delayed without affecting hemoglobin levels. Extending the proliferation of differentiating cells requires the synergistic action of CDK2 and CDK4. Importantly, CDK6 cannot substitute for CDK4 in this role, which demonstrates that the 2 cyclin D-dependent kinases are functionally different. The results show that differentiating hemoglobinized cells can be made to proliferate far beyond their normal capacity to divide. (Blood. 2000;96:2755-2764) PMID- 11023508 TI - Cell cycle exit during terminal erythroid differentiation is associated with accumulation of p27(Kip1) and inactivation of cdk2 kinase. AB - Progression through the mammalian cell cycle is regulated by cyclins, cyclin- dependent kinases (CDKs), and cyclin-dependent kinase inhibitors (CKIs). The function of these proteins in the irreversible growth arrest associated with terminally differentiated cells is largely unknown. The function of Cip/Kip proteins p21(Cip1) and p27(Kip1) during erythropoietin-induced terminal differentiation of primary erythroblasts isolated from the spleens of mice infected with the anemia-inducing strain of Friend virus was investigated. Both p21(Cip1) and p27(Kip1) proteins were induced during erythroid differentiation, but only p27(Kip1) associated with the principal G(1) CDKs-cdk4, cdk6, and cdk2. The kinetics of binding of p27(Kip1) to CDK complexes was distinct in that p27(Kip1) associated primarily with cdk4 (and, to a lesser extent, cdk6) early in differentiation, followed by subsequent association with cdk2. Binding of p27(Kip1) to cdk4 had no apparent inhibitory effect on cdk4 kinase activity, whereas inhibition of cdk2 kinase activity was associated with p27(Kip1) binding, accumulation of hypo-phosphorylated retinoblastoma protein, and G(1) growth arrest. Inhibition of cdk4 kinase activity late in differentiation resulted from events other than p27(Kip1) binding or loss of cyclin D from the complex. The data demonstrate that p27(Kip1) differentially regulates the activity of cdk4 and cdk2 during terminal erythroid differentiation and suggests a switching mechanism whereby cdk4 functions to sequester p27(Kip1) until a specified time in differentiation when cdk2 kinase activity is targeted by p27(Kip1) to elicit G(1) growth arrest. Further, the data imply that p21(Cip1) may have a function independent of growth arrest during erythroid differentiation. (Blood. 2000;96:2746-2754) PMID- 11023510 TI - A hematopoietic cell L-selectin ligand that is distinct from PSGL-1 and displays N-glycan-dependent binding activity. AB - Human hematopoietic progenitor cells express L-selectin and also express PSGL-1, a ligand for all selectins. Using a shear-based adhesion assay, a hematopoietic cell L-selectin ligand (HCLL) that is expressed on the hematopoietic cell line KG1a and on normal human hematopoietic progenitors was previously identified. To characterize the structural biology of HCLL and to define its relationship to PSGL-1, the effects of chemical and enzymatic treatments on HCLL activity of KG1a cells and membrane preparations were analyzed. Protease digestions and chemical treatments of KG1a cells and membranes indicated that HCLL is an integral membrane glycoprotein. Glycosidase digestions of membrane protein preparations and metabolic treatments of KG1a cells with glycosylation processing modifiers revealed that L-selectin binding determinants on HCLL are sialofucosylated structures presented on complex-type N-glycans. Adhesion assays and biochemical studies showed that this glycoprotein is also expressed on circulating blasts in native acute leukemias. HCLL is distinguishable from PSGL-1: (1) KG1a cells sorted for PSGL-1 expression had equivalent HCLL activity; (2) anti-PSGL-1 blocking antibodies and proteases known to eliminate L-selectin binding to PSGL-1 had no effect on HCLL binding activity of KG1a cells; (3) blasts from native leukemias with low expression of PSGL-1 and CD34 display high HCLL activity; and (4) despite high level expression of PSGL-1, HCLL activity was absent on HL60 cells. These data provide first evidence of a naturally expressed membrane L selectin ligand expressing binding determinant(s) on an N-linked glycoconjugate. This novel ligand may help mediate L-selectin-dependent cell-cell adhesive interactions within the cytoarchitecture of the bone marrow microenvironment. (Blood. 2000;96:2765-2774) PMID- 11023511 TI - Urokinase receptor expression on human microvascular endothelial cells is increased by hypoxia: implications for capillary-like tube formation in a fibrin matrix. AB - Hypoxia stimulates angiogenesis, the formation of new blood vessels. This study evaluates the direct effect of hypoxia (1% oxygen) on the angiogenic response of human microvascular endothelial cells (hMVECs) seeded on top of a 3-dimensional fibrin matrix. hMVECs stimulated with fibroblast growth factor-2 (FGF-2) or vascular endothelial growth factor (VEGF) together with tumor necrosis factor alpha (TNF-alpha) formed 2- to 3-fold more tubular structures under hypoxic conditions than in normoxic (20% oxygen) conditions. In both conditions the in growth of capillary-like tubular structures into fibrin required cell-bound urokinase-type plasminogen activator (uPA) and plasmin activities. The hypoxia induced increase in tube formation was accompanied by a decrease in uPA accumulation in the conditioned medium. This decrease in uPA level was completely abolished by uPA receptor-blocking antibodies. During hypoxic culturing uPA receptor activity and messenger RNA (mRNA) were indeed increased. This increase and, as a consequence, an increase in plasmin formation contribute to the hypoxia induced stimulation of tube formation. A possible contribution of VEGF-A to the increased formation under hypoxic conditions is unlikely because there was no increased VEGF-A expression detected under hypoxic conditions, and the hypoxia induced tube formation by FGF-2 and TNF-alpha was not inhibited by soluble VEGFR 1 (sVEGFR-1), or by antibodies blocking VEGFR-2. Furthermore, although the alpha(v)-integrin subunit was enhanced by hypoxia, blocking antibodies against alpha(v)beta(3)- and alpha(v)beta(5)-integrins had no effect on hypoxia-induced tube formation. Hypoxia increases uPA association and the angiogenic response of human endothelial cells in a fibrin matrix; the increase in the uPA receptor is an important determinant in this process. (Blood. 2000;96:2775-2783) PMID- 11023512 TI - Induction of decay-accelerating factor by thrombin through a protease-activated receptor 1 and protein kinase C-dependent pathway protects vascular endothelial cells from complement-mediated injury. AB - There is increasing evidence for functional crosstalk between inflammatory and thrombotic pathways in inflammatory vascular diseases such as atherosclerosis and vasculitis. Thus, complement activation on the endothelial cell (EC) surface during inflammation may generate thrombin via the synthesis of tissue factor. We explored the hypothesis that thrombin induces EC expression of the complement regulatory proteins decay-accelerating factor (DAF), membrane cofactor protein (MCP), and CD59 and that this maintains vascular integrity during coagulation associated with complement activation. Thrombin increased DAF expression on the surface of ECs by 4-fold in a dose- and time-dependent manner as measured by flow cytometry. DAF up-regulation was first detectable at 6 hours and maximal 24 hours poststimulation, whereas no up-regulation of CD59 or MCP was seen. Thrombin induced expression required increased DAF messenger RNA and de novo protein synthesis. The response depended on activation of protease-activated receptor 1 (PAR1) and was inhibited by pharmacologic antagonists of protein kinase C (PKC), p38 and p42/44 mitogen-activated protein kinase, and nuclear factor-kappa B. The increased DAF expression was functionally relevant because it significantly reduced C3 deposition and complement-mediated EC lysis. Thus, thrombin-generated at inflammatory sites in response to complement activation-is a physiologic agonist for the PKC-dependent pathway of DAF regulation, thereby providing a negative feedback loop protecting against thrombosis in inflammation. (Blood. 2000;96:2784-2792) PMID- 11023513 TI - Analysis of fibrin formation and proteolysis during intravenous administration of ancrod. AB - Ancrod is a purified fraction of venom from the Malayan pit viper, Calloselasma rhodostoma, currently under investigation for treatment of acute ischemic stroke. Treatment with ancrod leads to fibrinogen depletion. The present study investigated the mechanisms leading to the reduction of plasma fibrinogen concentration. Twelve healthy volunteers received an intravenous infusion of 0.17 U/kg body weight of ancrod for 6 hours. Blood samples were drawn and analyzed before and at various time points until 72 hours after start of infusion. Ancrod releases fibrinopeptide A from fibrinogen, leading to the formation of desAA fibrin monomer. In addition, a considerable proportion of desA-profibrin is formed. Production of desA-profibrin is highest at low concentrations of ancrod, whereas desA-profibrin is rapidly converted to desAA-fibrin at higher concentrations of ancrod. Both desA-profibrin and desAA-fibrin monomers form fibrin complexes. A certain proportion of complexes carries exposed fibrin polymerization sites E(A), indicating that the terminal component of the protofibril is a desAA-fibrin monomer unit. Soluble fibrin complexes potentiate tissue-type plasminogen activator-induced plasminogen activation. Significant amounts of plasmin are formed when soluble fibrin in plasma reaches a threshold concentration, leading to the proteolytic degradation of fibrinogen and fibrin. In the present setting, high concentrations of soluble fibrin are detected after 1 hour of ancrod infusion, whereas a rise in fibrinogen and fibrin degradation products, and plasmin-alpha(2)-plasmin inhibitor complex levels is first detected after 2 hours of ancrod infusion. Ancrod treatment also results in the appearance of cross-inked fibrin degradation product D-dimer in plasma. (Blood. 2000;96:2793 2802) PMID- 11023514 TI - A partial deficiency of interleukin-7R alpha is sufficient to abrogate T-cell development and cause severe combined immunodeficiency. AB - Both in vitro and in vivo studies established that interleukin 7 (IL-7) is essential for differentiation of immature T cells and B cells but not natural killer (NK) cells in the mouse. In humans, although both T-cell and B-cell progenitors express the functional IL-7 receptor that consists of IL-7R alpha and the gamma common (gamma c) chain, this lymphocyte receptor system is critical for T lineage but not for B lineage development. Indeed, complete gamma c deficiency like IL-7R alpha deficiency results in the arrest of T-cell but not B-cell development (T(-)B(+) SCID). However, partial deficiency of gamma c caused by missense mutations results in a T(+)B(+) phenotype and a delay of clinical presentation. It was therefore plausible to assume that partial deficiency of IL 7R alpha, like partial gamma c deficiency may lead to a milder clinical and immunologic phenotype. A P132S mutation in the IL-7R alpha was identified in 3 patients with severe combined immunodeficiency (SCID) within an extensively consanguineous family. Substitution of proline with serine in the extracellular portion of IL-7R alpha did not affect IL-7R alpha messenger RNA (mRNA) and protein expression, but severely compromised affinity to IL-7, resulting in defective signal transduction. In response to IL-7 stimulation, Jak-3 phosphorylation was markedly reduced in both patient cells as well as in COS cells reconstituted with mutant IL-7R alpha. Surprisingly, this partial deficiency of IL-7R alpha resulted in a severe phenotype, including markedly reduced circulating T cells while sparing B-cell numbers similar to gamma c chain deficiency. However, unlike the previously reported cases, serum immunoglobulins were virtually absent. Further, unlike gamma c deficiency, NK cell numbers and function was preserved. Despite the partial deficiency, clinical presentation was indistinguishable from a complete gamma c deficiency, including severe and persistent viral and protozoal infections and failure to thrive. Unlike partial gamma c deficiency, a partial deficiency of IL-7R alpha results in an arrest of T cell development, leading to typical severe combined immunodeficiency. This underscores the critical role of IL-7R alpha chain in the differentiation of T cells. (Blood. 2000;96:2803-2807) PMID- 11023515 TI - Costimulation of T cells by B7-H2, a B7-like molecule that binds ICOS. AB - This report describes a new human B7-like gene designated B7-H2. Cell surface expression of B7-H2 protein is detected in monocyte-derived immature dendritic cells. Soluble B7-H2 and immunoglobulin (Ig) fusion protein, B7-H2Ig, binds activated but not resting T cells and the binding is abrogated by inducible costimulator Ig (ICOSIg), but not CTLA4Ig. In addition, ICOSIg stains Chinese hamster ovary cells transfected with B7-H2 gene. By suboptimal cross-linking of CD3, costimulation of T-cell proliferation by B7-H2Ig is dose-dependent and correlates with secretion of interleukin (IL)-2, whereas optimal CD3 ligation preferentially stimulates IL-10 production. The results indicate that B7-H2 is a putative ligand for the ICOS T-cell molecule. (Blood. 2000;96:2808-2813) PMID- 11023516 TI - Rapid reconstitution of Epstein-Barr virus-specific T lymphocytes following allogeneic stem cell transplantation. AB - Epstein-Barr virus (EBV)-specific CD8 T lymphocytes are present at remarkably high frequencies in healthy EBV(+) individuals and provide protection from EBV associated lymphoproliferative diseases. Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) is a commonly used therapy in which T-cell surveillance for EBV is temporarily disrupted. Herein, human leukocyte antigen (HLA) class I tetramers were used to investigate the reestablishment of the EBV specific CD8 T-cell repertoire in patients following allo-PBSCT. CD8(+) T cells specific for lytic and latent cycle-derived EBV peptides rapidly repopulate the periphery of matched sibling allo-PBSCT patients. The relative frequencies of T cells specific for different EBV peptides in transplantation recipients closely reflect those of their respective donors. Investigation of patients at monthly intervals following unmanipulated allo-PBSCT demonstrated that the frequency of EBV-specific T cells correlates with the number of EBV genome copies in the peripheral blood and that expansion of EBV-specific T-cell populations occurs even in the setting of immunosuppressive therapy. In contrast, patients undergoing T-cell-depleted or unrelated cord blood transplantation have undetectable EBV-specific T cells, even in the presence of Epstein-Barr viremia. The protective shield provided by EBV-specific CD8 T cells is rapidly established following unmanipulated matched sibling allo-PBSCT and demonstrates that HLA class I tetramers complexed with viral peptides can provide direct and rapid assessment of pathogen-specific immunity in this and other vulnerable patient populations. (Blood. 2000;96:2814-2821) PMID- 11023517 TI - Induction of neutrophil responsiveness to myeloperoxidase antibodies by their exposure to supernatant of degranulated autologous neutrophils. AB - Antibodies against myeloperoxidase (MPO) and proteinase 3 (PR3) are the predominant autoantibodies present in antineutrophil cytoplasmic antibody (ANCA) associated vasculitis. Their binding to the corresponding antigen on the surface of polymorphonuclear neutrophils (PMNs) is believed to trigger the disease process. Cytokines released during an inflammatory reaction are thought to prime resting PMNs, making them responsive to autoantibodies. In the present study we found that MPO but not PR3 could be detected on the cell surface of unstimulated PMNs after incubation with the supernatants of activated autologous PMNs. MPO was shown to be acquired from these supernatants, because PMNs did not express MPO when the supernatants were specifically MPO-depleted. In addition, purified soluble MPO bound to unstimulated PMNs. Unstimulated PMNs that had passively acquired MPO released oxygen radicals when incubated with monoclonal antibody anti-MPO or the immunoglobulin G fraction of a patient with MPO-ANCA. The data presented here suggest that, in ANCA-associated vasculitis, soluble MPO released by activated PMNs may bind to unstimulated PMNs, thereby making them reactive to anti-MPO antibodies. This mechanism of dispersing PMN activation would be specific for MPO-ANCA and may explain differences in the pathologic and clinical expression of MPO-ANCA versus PR3-ANCA vasculitis. (Blood. 2000;96:2822-2827) PMID- 11023518 TI - Tumor-specific recognition of human myeloma cells by idiotype-induced CD8(+) T cells. AB - Immunoglobulin secreted by myeloma cells contains a unique antigenic determinant (idiotype [Id]) that may serve as a tumor-specific antigen. Although Id-protein specific T-cell responses have been reported in patients with myeloma, it is not known whether primary myeloma tumor cells can present naturally processed Id peptides on their surface as a target. We immunized 2 healthy human stem-cell donors with Id proteins from their recipients. T cells from the immunized donors released high levels of T-helper 1-type cytokines in response to stimulation with myeloma cells from their recipients. The T-cell-mediated cytokine response to tumor cells was blocked by a major histocompatibility complex (MHC) class I monoclonal antibody, whereas the response to soluble Id protein was dependent on MHC class II. To investigate whether Id-specific CD8(+) T cells can recognize and kill autologous myeloma cells, we generated T cells from peripheral blood mononuclear cells from a third patient with myeloma by means of in vitro stimulation with autologous dendritic cells pulsed with Id protein. Tumor specific lysis of myeloma cells was demonstrated by the lack of killing of autologous nonmalignant B cells or natural killer-sensitive K562 cells. Lysis of autologous myeloma targets was restricted by MHC class I molecules. These data represent the first report of class I-restricted T-cell recognition of fresh autologous myeloma targets and formally demonstrate that human myeloma cells can serve as targets of an Id-specific T-cell response. (Blood. 2000;96:2828-2833) PMID- 11023519 TI - Treatment with liposome-encapsulated clodronate as a new strategic approach in the management of immune thrombocytopenic purpura in a mouse model. AB - Immune thrombocytopenic purpura (ITP) is an autoimmune disease related to the presence of elevated levels of platelet-associated immunoglobulin, or autoantibodies. In recent years the importance of macrophage Fc gamma receptors in the uptake of platelets in ITP has been confirmed. Although in patients with ITP the platelet destruction occurs in liver and spleen, in this present experimental mouse model the liver was the principal organ of sequestration of sensitized platelets. The uptake in the spleen, bone marrow, lung, and kidneys was negligible and not different from that in control animals. In addition, the trapped platelets did not return to circulation, and new cells derived from the platelet-storage pool or new thrombocytogenesis were necessary to restore the platelet count. The depletion of splenic and hepatic murine macrophages by liposome-encapsulated clodronate (lip-clod) was studied as a new strategy for ITP treatment. Lip-clod inhibits, in a dose-dependent manner, the antibody-induced thrombocytopenia. Moreover, lip-clod treatment rapidly restored (24 hours) the platelet count in thrombocytopenic animals to hematologic safe values, and despite additional antiplatelet antiserum treatment, mice were able to maintain this level of platelets at least up to 48 hours. The bleeding times in lip-clod treated animals was not different from those in controls, demonstrating that the hemostasis was well controlled in these animals. The results presented in this study demonstrate that lip-clod treatment can be effective in the management of experimental ITP. (Blood. 2000;96:2834-2840) PMID- 11023520 TI - Hodgkin disease: pharmacologic intervention of the CD40-NF kappa B pathway by a protease inhibitor. AB - The malignant Reed-Sternberg cell of Hodgkin disease is an aberrant B cell that persists in an immunolgically mediated inflammatory infiltrate. Despite its nonproductive immunoglobulin genes, the Reed-Sternberg cell avoids the usual apoptotic fate of defective immune cells through an unknown mechanism. A likely candidate is the surface receptor, CD40, consistently expressed by Reed-Sternberg cells, and the first link in the pathway to NF-kappa B activation, the central regulator of cytokine production and apoptosis. CD40 signaling in B lymphocytes coordinates the immune response, including immunoglobulin isotype switch and Fas mediated apoptosis. CD40-induced NF-kappa B activation is mediated by adapter proteins, the TNF receptor (TNFR)-associated factors (TRAFs), especially TRAFs 2, 3, and 5. Using a Hodgkin cell line, this study demonstrates that CD40 activation of NF-kappa B is mediated by proteolysis of TRAF3. Results further demonstrate that the pathway can be blocked by treatment with pharmacologic doses of a specific protease inhibitor, pepstatin-A, even in the presence of a mutated NF kappa B inhibitor, I-kappa B alpha. The stability of TRAF3 regulates CD40/NF kappa B-mediated control of the immune response, which is central to the biologic activity of the Reed-Sternberg cell. Prevention of TRAF3 proteolysis may be an entry point for design of novel pharmaceuticals to treat Hodgkin disease and immune system disorders. (Blood. 2000;96:2841-2848) PMID- 11023521 TI - Arsenic-interferon-alpha-triggered apoptosis in HTLV-I transformed cells is associated with tax down-regulation and reversal of NF-kappa B activation. AB - Human T-cell lymphotropic virus type I (HTLV-I)-associated adult T-cell leukemia/lymphoma (ATL) is a malignancy of mature activated T cells resistant to conventional chemotherapy. The viral transactivator protein Tax plays a critical role in HTLV-I-induced transformation and apoptosis resistance by inducing I kappa B-alpha degradation, resulting in the activation of the NF-kappa Bpathway. In these HTLV-I-transformed cells, arsenic trioxide (As) and interferon (IFN) alpha synergize to induce cell cycle arrest and apoptosis. We demonstrate that cell death induction is only partly dependent upon caspase activation and is not associated with modulation of bcl-2, bax, or p53 expression. However, combined As and IFN induce the degradation of Tax, associated with an up-regulation of I kappa B-alpha resulting in a sharp decrease in RelA DNA binding nuclear factor (NF)-kappa B complexes because of the cytoplasmic retention of RelA. Taken the role of Tax in HTLV-I-induced transformation, its down-regulation probably accounts for cell death induction through inactivation of the NF-kappa B pathway. Such specific targeting of the viral oncoprotein by As-IFN treatment, reminiscent of As targeting of promyelocytic leukemia/retinoic acid receptor-alpha in acute promyelocytic leukemia, provides strong rational for combined As-IFN therapy in ATL patients. (Blood. 2000;96:2849-2855) PMID- 11023522 TI - Insulin-like growth factor I is a dual effector of multiple myeloma cell growth. AB - Multiple myeloma (MM) is an invariably fatal disease that accounts for approximately 1% to 2% of all human cancers. Surprisingly little is known about the cellular pathways contributing to growth of these tumors. Although the cytokine interleukin-6 has been suggested to be the major stimulus for myeloma cell growth, the role of a second potential growth factor, insulin-like growth factor I (IGF-I), has been less clearly defined. The IGF-I signaling cascade in 8 MM cell lines was examined. In 7 of these, the IGF-I receptor (IGF-IR) was expressed and autophosphorylated in response to ligand. Downstream of IGF-IR, insulin receptor substrate 1 was phosphorylated, leading to the activation of phosphatidylinositol-3'-kinase (PI-3K). PI-3K, in turn, regulated 2 distinct pathways. The first included Akt and Bad, leading to an inhibition of apoptosis; the second included the mitogen-activated protein kinase (MAPK), resulting in proliferation. Biologic relevance of this pathway was demonstrated because in vitro IGF-I induced both an antiapoptotic and a proliferative effect. Importantly, in vivo administration of IGF-I in SCID mice inoculated with the OPM 2 line led to approximately twice the growth rate of tumor cells as in controls. These results suggest that IGF-I activates at least 2 pathways effecting myeloma cell growth and contributes significantly to expansion of these cells in vivo. (Blood. 2000;96:2856-2861) PMID- 11023523 TI - High incidence of biallelic point mutations in the Runt domain of the AML1/PEBP2 alpha B gene in Mo acute myeloid leukemia and in myeloid malignancies with acquired trisomy 21. AB - The AML1 gene, situated in 21q22, is often rearranged in acute leukemias through t(8;21) translocation, t(12;21) translocation, or less often t(3;21) translocation. Recently, point mutations in the Runt domain of the AML1 gene have also been reported in leukemia patients. Observations for mutations of the Runt domain of the AML1 gene in bone marrow cells were made in 300 patients, including 131 with acute myeloid leukemia (AML), 94 with myelodysplastic syndrome (MDS), 28 with blast crisis chronic myeloid leukemia (CML), 3 with atypical CML, 41 with acute lymphoblastic leukemia (ALL), and 3 with essential thrombocythemia (ET). Forty-one of the patients had chromosome 21 abnormalities, including t(8;21) in 6 of the patients with AML, t(12;21) in 8 patients with ALL, acquired trisomy 21 in 17 patients, tetrasomy 21 in 7 patients, and constitutional trisomy 21 (Down syndrome) in 3 patients. A point mutation was found in 14 cases (4.7%), including 9 (22%) of the 41 patients with AML of the Mo type (MoAML) (none of them had detectable chromosome 21 rearrangement) and 5 (38%) of the 13 myeloid malignancies with acquired trisomy 21 (1 M1AML, 2 M2AML, 1 ET, and 1 atypical CML). In at least 8 of 9 mutated cases of MoAML, both AML alleles were mutated: 3 patients had different stop codon mutations of the 2 AML1 alleles, and 5 patients had the same missense or stop codon mutation in both AML1 alleles, which resulted in at least 3 of the patients having duplication of the mutated allele and deletion of the normal residual allele, as shown by FISH analysis and by comparing microsatellite analyses of several chromosome 21 markers on diagnosis and remission samples. In the remaining mutated cases, with acquired trisomy 21, a missense mutation of AML1, which involved 2 of the 3 copies of the AML1 gene, was found. Four of the 7 mutated cases could be reanalyzed in complete remission, and no AML1 mutation was found, showing that mutations were acquired in the leukemic clone. In conclusion, these findings confirm the possibility of mutations of the Runt domain of the AML1 gene in leukemias, mainly in MoAML and in myeloid malignancies with acquired trisomy 21. AML1 mutations, in MoAML, involved both alleles and probably lead to nonfunctional AML1 protein. As AML1 protein regulates the expression of the myeloperoxidase gene, the relationship between AML1 mutations and Mo phenotype in AML will have to be further explored. (Blood. 2000;96:2862-2869) PMID- 11023524 TI - Phosphorylation-deficient Stat1 inhibits retinoic acid-induced differentiation and cell cycle arrest in U-937 monoblasts. AB - All-trans retinoic acid (ATRA) is a potent inducer of terminal differentiation of immature leukemic cell lines in vitro and of acute promyelocytic leukemia (APL) cells in vivo. Recent reports have shown that ATRA induces the expression of several interferon-regulated genes, including signal transducer and activator of transcription (Stat)1. To investigate the role of Stat1 activation in ATRA signaling, sublines were established for the human monoblastic cell line U-937 constitutively expressing wild-type or phosphorylation-defective Stat1, mutated in the conserved tyrosine 701 required for dimerization and nuclear translocation. Results showed that ATRA induction leads to activation of Stat1 by the phosphorylation of tyrosine 701 and subsequent nuclear translocation. Consistent with a functional importance of this activation, ectopic expression of Stat1(Y701F) suppressed ATRA-induced morphologic differentiation and expression of the monocytic surface markers CD11c and the granulocyte colony-stimulating factor receptor. Moreover, ATRA-induced growth arrest in the G(0)/G(1) phase of the cell cycle was inhibited by phosphorylation-deficient Stat1. Taken together, these results indicate that Stat1 is a key mediator of ATRA-induced cell cycle arrest and differentiation of U-937 cells. (Blood. 2000;96:2870-2878) PMID- 11023525 TI - Cellular drug resistance profiles in childhood acute myeloid leukemia: differences between FAB types and comparison with acute lymphoblastic leukemia. AB - Determining in vitro drug resistance may reveal clinically relevant information in childhood leukemia. Using the methyl-thiazol-tetrazolium assay, the resistance of untreated leukemic cells to 21 drugs was compared in 128 children with acute myeloid leukemia (AML) and 536 children with acute lymphoblastic leukemia (ALL). The differences between 3 French-American-British (FAB) types (M1/M2, M4, and M5) were also compared. AML was significantly more resistant than ALL to the following drugs, as noted by the median resistance: glucocorticoids (greater than 85-fold), vincristine (4.4-fold), L-asparaginase (6.9-fold), anthracyclines (1.8- to 3.4-fold), mitoxantrone (2.6-fold), etoposide (4.9-fold), platinum analogues (2.4- to 3.4-fold), ifosfamide (3.5-fold), and thiotepa (3.9-fold). For cytarabine and thiopurines, the median LC50 values (the drug concentration that kills 5% of the cells) were equal. Also, busulfan, amsacrine, teniposide, and vindesine showed no significant differences, but the numbers were smaller, and the median LC50 values were 1.3- to 5.2-fold higher in AML. None of the drugs demonstrated greater cytotoxicity in AML. FAB M5 was significantly more sensitive than FAB M4 to most drugs frequently used in AML, as indicated by the following ratios of median sensitivities: the anthracyclines (2.6- to 3.2-fold), mitoxantrone (12.5-fold), etoposide (8.7-fold), and cytarabine (2.9-fold). For etoposide and cytarabine (5.4- and 3.4-fold, respectively) FAB M5 was also significantly more sensitive than FAB M1/M2. FAB M5 was equally sensitive to L asparaginase and vincristine as ALL. Only 15% of the AML samples were "intermediately" sensitive to glucocorticoids, mainly in FAB M1/M2. The poorer prognosis of childhood AML is related to resistance to a large number of drugs. Within AML, FAB M5 had a distinct resistance pattern. These resistance profiles may be helpful in the rational design of further treatment protocols. (Blood. 2000;96:2879-2886) PMID- 11023526 TI - Down-regulation of the chemokine receptor CCR5 by activation of chemotactic formyl peptide receptor in human monocytes. AB - Interactions between cell surface receptors are important regulatory elements in the complex host responses to infections. In this study, it is shown that a classic chemotactic factor, the bacterial chemotactic peptide N-formyl-methionyl leucylphenyl-alanine (fMLF), rapidly induced a protein-kinase-C-mediated serine phosphorylation and down-regulation of the chemokine receptor CCR5, which serves as a major human immunodeficiency virus (HIV)-1 coreceptor. The fMLF binding to its receptor, formyl peptide receptor (FPR), resulted in significant attenuation of cell responses to CCR5 ligands and in inhibition of HIV-1-envelope glycoprotein-mediated fusion and infection of cells expressing CD4, CCR5, and FPR. The finding that the expression and function of CCR5 can be regulated by peptides that use an unrelated receptor may provide a novel approach to the design of anti-inflamatory and antiretroviral agents. (Blood. 2000;96:2887-2894) PMID- 11023527 TI - Nonsense mutations in the human beta-globin gene lead to unexpected levels of cytoplasmic mRNA accumulation. AB - Generally, nonsense codons 50 bp or more upstream of the 3'-most intron of the human beta-globin gene reduce mRNA abundance. In contrast, dominantly inherited beta-thalassemia is frequently associated with nonsense mutations in the last exon. In this work, murine erythroleukemia (MEL) cells were stably transfected with human beta-globin genes mutated within each of the 3 exons, namely at codons 15 (TGG-->TGA), 39 (C-->T), or 127 (C-->T). Primer extension analysis after erythroid differentiation induction showed codon 127 (C-->T) mRNA accumulated in the cytoplasm at approximately 20% of the normal mRNA level. Codon 39 (C-->T) mutation did not result in significant mRNA accumulation. Unexpectedly, codon 15 (TGG-->TGA) mRNA accumulated at approximately 90%. Concordant results were obtained when reticulocyte mRNA from 2 carriers for this mutation was studied. High mRNA accumulation of codon 15 nonsense-mutated gene was revealed to be independent of the type of nonsense mutation and the genomic background in which this mutation occurs. To investigate the effects of other nonsense mutations located in the first exon on the mRNA level, nonsense mutations at codons 5, 17, and 26 were also cloned and stably transfected into MEL cells. After erythroid differentiation induction, mRNAs with a mutation at codon 5 or 17 were detected at high levels, whereas the mutation at codon 26 led to low mRNA levels. These findings suggest that nonsense-mediated mRNA decay is not exclusively dependent on the localization of mutations relative to the 3'-most intron. Other factors may also contribute to determine the cytoplasmic nonsense-mutated mRNA level in erythroid cells. (Blood. 2000;96:2895-2901) PMID- 11023528 TI - CFU-S(11) activity does not localize solely with the aorta in the aorta-gonad mesonephros region. AB - The aorta-gonad-mesonephros (AGM) region is a potent hematopoietic site in the midgestation mouse conceptus and first contains colony-forming units-spleen day 11 (CFU-S(11)) at embryonic day 10 (E10). Because CFU-S(11) activity is present in the AGM region before the onset of hematopoietic stem cell (HSC) activity, CFU S(11) activity in the complex developing vascular and urogenital regions of the AGM was localized. From E10 onward, CFU-S(11) activity is associated with the aortic vasculature, and is found also in the urogenital ridges (UGRs). Together with data obtained from organ explant cultures, in which up to a 16-fold increase in CFU-S(11) activity was observed, it was determined that CFU-S(11) can be increased autonomously both in vascular sites and in UGRs. Furthermore, CFU-S(11) activity is present in vitelline and umbilical vessels. This, together with the presence of CFU-S(11) in the UGRs 2 days before HSC activity, suggests both temporally and spatially distinct emergent sources of CFU-S(11). (Blood. 2000;96:2902-2904) PMID- 11023529 TI - De novo factor VIII gene intron 22 inversion in a female carrier presents as a somatic mosaicism. AB - The intron 22 inversion represents the most prevalent factor VIII gene defect in severe hemophilia A, accounting for about 40% of all mutations. It is hypothesized that the inversion mutations occur almost exclusively in germ cells during meiotic cell division by intrachromosomal recombination between 1 of 2 telomeric copies of the Int22h region and its intragenic homologue. The majority of inversion mutations originate in male germ cells, where the lack of bivalent formation may facilitate flipping of the telomeric end of the single X chromosome. This is the first intron 22 inversion that presents as a somatic mosaicism in a female, affecting only about 50% of lymphocyte and fibroblast cells of the proposita. Supposing a post-zygotic de novo mutation as the usual cause of somatic mosaicism, the finding would imply that the intron 22 inversion mutation is not restricted to meiotic cell divisions but can also occur during mitotic cell divisions, either in germ cell precursors or in somatic cells. (Blood. 2000;96:2905-2906) PMID- 11023530 TI - Nonimmunoglobulin (non-Ig)/BCL6 gene fusion in diffuse large B-cell lymphoma results in worse prognosis than Ig/BCL6. AB - Chromosomal translocation involving the BCL6 gene affects not only immunoglobulin (Ig) genes but also a number of non-Ig genes as partners. The molecular anatomy of the BCL6 gene rearrangements in 39 cases with diffuse large B-cell lymphoma (DLBCL) by long-distance polymerase chain reaction-based assays was determined. The results showed that Ig genes were affected in 21 cases; non-Ig genes, 15 cases; a deletion of more than a 1-kb segment, 2 cases; and a point mutation, 1 case. Comparative studies between the 21 cases with Ig gene partners and the 17 cases with non-Ig gene partners, including 2 cases with the deletion, showed that the overall survival of the latter group of patients was significantly inferior to that of the former (P = .0440), and the estimated 2-year overall survival rates were 58.3% vs 17.6% (P = .005). Non-Ig/BCL6 fusion is a poor prognostic indicator of DLBCL, and DLBCL with BCL6 translocation could be subclassified according to the individual partner locus and/or gene. (Blood. 2000;96:2907-2909) PMID- 11023531 TI - Bcl-2 rearrangement in patients with chronic hepatitis C associated with essential mixed cryoglobulinemia type II. AB - Hepatitis C virus (HCV) infection is found in 80% to 90% of patients with essential mixed cryoglobulinemia (EMC) type II, which is associated with monoclonal IgMk produced by monoclonal B cells. It was investigated whether bcl-2 rearrangement is associated with the clonal B-cell proliferation of EMC induced by hepatitis C. The study groups were composed of 15 patients with HCV and EMC, 12 patients with HCV without EMC, and 7 patients with chronic liver disease (CLD) unrelated to HCV. Fluorescence in situ hybridization with probes was applied to JH and to bcl-2 to study whether JH/bcl-2 translocation was present in these patients. Thirteen of 15 (86%) of patients with HCV-related EMC had the JH/bcl-2 translocation, a significantly higher rate than in HCV patients without EMC (16%; P < .001). Bcl-2 rearrangement was not detected in the patients with CLD not related to HCV. The JH/bcl-2 translocation may constitute a pathogenetic link for the development of NHL in patients with HCV infection. (Blood. 2000;96:2910-2912) PMID- 11023532 TI - The origin of oncogenic mutations: where is the primary damage? AB - Cancer is generally believed to arise from a single cell which has become 'initiated' by mutation of a few crucial genes, caused by random 'hits' in its DNA, a 'hit' being an error in DNA replication or a reaction of the DNA with free radicals or other chemical species of exogenous or endogenous origin. It is not obvious how the epidemiological data on cancer incidence can be interpreted within the framework of this paradigm. For example, it cannot account quantitatively for the age dependence of cancer incidence, or for the fact that the incidence of cancer in people with hereditary mutations in tumour suppressor genes is much lower than expected, or for the observation that while in some types of cancer, like colon and pancreas, certain highly oncogenic mutations, such as that of TP53, are prevalent, there is no significant increase in the incidence of these cancers in people who carry the mutations by heredity. It is argued here that although mutations in such genes appear to be of crucial importance in carcinogenesis they may not be the rate limiting events in common cancer. The epidemiological data are consistent with the hypothesis that the rate limiting processes involve large numbers of cells. Conceivably, the mutations directly underlying neoplastic transformation may be the result of a local collapse in the system of intercellular processes on which the stability of the normal genotype and phenotype depends, and thereby trigger a cascade of mutations, among them the highly oncogenic ones. This local collapse may be due to mutations of many different genes in many cells as well as to other factors affecting the integrity of tissue. PMID- 11023533 TI - Concordant induction of 15-lipoxygenase-1 and mutant p53 expression in human prostate adenocarcinoma: correlation with Gleason staging. AB - We recently reported that the mutant form of the tumor-suppressor gene p53 up regulates 15-LO-1 gene expression in a murine cell line. Here, we examine the expression of 15-lipoxygenase (LO)-1 and mutant p53 (mtp53) in human prostatic tissues and 15-LO-1 in the human prostate adenocarcinoma cell line PC-3. Reverse transcription-PCR and western analyses conclusively demonstrated expression of 15 LO-1 in PC-3 cells. Western blotting for 15-LO-1 in freshly resected 'normal' and prostate adenocarcinoma specimens showed 15-LO-1 expression in normal tissue, but significantly higher levels were detected in prostate adenocarcinomas. Prostate adenocarcinoma tissues generated chirally pure 13-S-hydroxyoctadecadienoic acid from exogenous linoleic acid, a preferred substrate of 15-LO-1. To study the correlation of 15-LO-1 expression with mtp53 in prostate cancer, we immunostained 48 prostatectomy specimens obtained by transurethral resection of the prostate and needle biopsy (median age 68 years, range 52-93) of different Gleason grades (n = 48), using antibodies specific for 15-LO-1, mtp53 and MIB-1 (a proliferation marker). We compared staining in cancerous foci with adjacent normal appearing prostate tissues. In only 5 of 48 patients did 'normal' tissue adjacent to cancerous foci display staining for 15-LO-1. However, no staining for mtp53 was observed in any of the normal tissues. In cancer foci, robust staining was observed for both 15-LO-1 (36 of 48, 75%) and mtp53 (19 of 48, 39%). Furthermore, the intensities of expression of 15-LO-1 and mtp53 correlated positively with each other (P < 0.001) and with the degree of malignancy, as assessed by Gleason grading (P < 0.01). By immunohistochemistry, 15-LO-1 was located in secretory cells of peripheral zone glands, prostatic ducts and seminal vesicles, but not in the basal cell layer or stroma. Based on these and other studies, we propose a model describing a possible role for 15-LO-1 expression in influencing the malignant potential and pathobiological behavior of adenocarcinomas. PMID- 11023534 TI - Cholinergic receptor up-regulates COX-2 expression and prostaglandin E(2) production in colon cancer cells. AB - The M(3) muscarinic cholinergic receptor has important physiological functions on normal colonic cells. It is frequently expressed on human colon cancer cells and is biologically active. Although it is mitogenic in certain cell models, the importance of this receptor on colon carcinogenesis is unknown. In the present study we have determined expression of the M(3) receptor on human colon cancer tissue compared with matched normal tissue and examined the downstream effect of receptor activation in the HT-29 human colon carcinoma cell line. Using reverse transcription-PCR, M(3) receptor RNA expression was detected in all matched colon carcinoma and normal specimens from eight patients. Five of the eight (62%) patients showed an up to 8-fold greater level of M(3) receptor expression in cancer compared with the matched normal tissue. Exposure of HT-29 cells to carbachol, a stable receptor agonist, results in a 10-fold increase in cyclooxygenase-2 (COX-2) protein. This induction of COX-2 protein was dose dependent and was inhibited by the cholinergic receptor antagonist N methylscopolamine (NMS). Carbachol caused a dose-dependent increase in prostaglandin E(2) (PGE(2)), the main product of cyclooxygenase activity. The maximum stimulatory effect (40-fold increase) was noted with 1mM carbachol. The increase in PGE(2) was completely abolished by NMS and by the COX-2 selective inhibitor NS398. This suggests that the M(3) receptor mediates PGE(2) production by a mechanism involving COX-2. As COX-2 and PGE(2) are known promoters of gastrointestinal cancer, these data suggest that M(3) receptor activation may facilitate progression of colon carcinoma, in part by a COX-2-mediated cellular mechanism. PMID- 11023535 TI - Induction of heme oxygenase-1 and adaptive protection against the induction of DNA damage after hyperbaric oxygen treatment. AB - Hyperbaric oxygen (HBO) treatment of human subjects (i.e. exposure to 100% oxygen at a pressure of 2.5 ATA for a total period of 3 x 20 min) caused clear and reproducible DNA damage in lymphocytes, as detected with the comet assay (single cell gel electrophoresis). Induction of DNA damage was found only after the first HBO exposure and not after further treatments of the same individuals. Furthermore, blood taken 24 h after HBO treatment was significantly protected against the induction of DNA damage by hydrogen peroxide (H(2)O(2)) in vitro, indicating that adaptation occurred due to induction of antioxidant defenses. The cells were not significantly protected against the genotoxic effects of gamma irradiation, suggesting increased scavenging of reactive oxygen species distant from nuclear DNA or an inducible change in the levels of free transition metals. We now demonstrate increased levels of heme oxygenase-1 (HO-1) in lymphocytes 24 h after HBO treatment of volunteers. Under the same conditions, superoxide dismutase, catalase and the DNA repair enzymes apurinic endonuclease and DNA polymerase beta were not enhanced in expression. We also show that protection against the induction of DNA damage by H(2)O(2) in lymphocytes even occurs with a shortened HBO treatment which did not induce significant DNA damage by itself. Our results suggest that increased sequestration of iron as a consequence of induced HO-1 might be involved in the adaptive protection after HBO treatment and that the induction of DNA damage is not the trigger for adaptive protection. PMID- 11023536 TI - Enhanced tumor growth in UV-irradiated skin is associated with an influx of inflammatory cells into the epidermis. AB - UV radiation causes a number of cellular changes within the skin which play a role in tumor outgrowth, including immunosuppression and production of growth enhancing cytokines. Both of these enable tumors to grow but their relative importance in carcinogenesis is poorly defined. In this study, C3H/HeN mice were exposed to a single inflammatory dose of 410 mJ/cm(2) UVB radiation (plus 100 mJ/cm(2) UVA radiation) followed by the inoculation of a regressor squamous cell carcinoma into or the painting of oxazolone onto the treated skin. Tumors transplanted 2 or 3 but not 4 days after irradiation had a significantly higher growth rate than tumors inoculated into unirradiated control mice. In contrast, mice failed to respond to hapten when it was applied 2, 3 or 4 days after irradiation. Cytofluorimetric analysis demonstrated that the number of F4/80(+) Langerhans cells was not significantly reduced until 4 days after irradiation, while the number of dendritic epidermal T cells was significantly lower at all time points observed after UV-irradiation. Furthermore, a large cellular infiltration of CD11b(+), Gr-1(+), CD45(+) MHC class II(+) and CD45(+) MHC class II(-) cells into the epidermis was observed 2 and 3 days after irradiation, which corresponded with the enhanced tumor growth. To a lesser extent tumor growth was also associated with CD45(+) MHC class II(hi) cells, possibly the previously described UV-induced macrophage. In contrast, suppression of contact hypersensitivity corresponded with the reduction in dendritic epidermal T cells but not with other cell changes. The results suggest that, in this model, where immunosuppression did not appear to be responsible for enhanced tumor growth, inflammatory infiltrates may contribute to the promotion of skin tumor growth within UV-irradiated skin. PMID- 11023537 TI - Proliferative potential in nasopharyngeal carcinoma: correlations with metallothionein expression and tissue zinc levels. AB - Metallothionein (MT) is a cysteine-rich protein with pleiotropic functions and a high binding affinity for heavy metals. The present study was designed to examine the relationship between MT expression and tissue zinc levels in conjunction with cell proliferation in nasopharyngeal cancer (NPC). Proliferative activity in NPC was quantified by Ki67 immunolabelling and MT expression was determined by immunohistochemistry. Total zinc and subcellular zinc fractions were analysed by flame atomic absorption spectrometry. MT immunostaining was observed in the nuclei of NPC cells, with the percentage MT immunopositivity ranging from 3.0 to 59.7%. Thirteen tumours displayed weak MT staining and the remaining 11 showed moderate to strong immunostaining. There was a significant positive correlation between MT and Ki67 positivity (P = 0.0127). Tissue zinc levels were higher in NPC as compared with benign nasopharyngeal tissues (4.800 +/- 0.4610 versus 2.889 +/- 0.4045 microgram/g dry wt tissue, respectively; P = 0.0122). Nuclear zinc levels in NPC were significantly higher than levels in membrane and cytosolic fractions (mean zinc levels 1.4840 +/- 0.1489, 0.6286 +/- 0.0789 and 0.3014 +/- 0.0250 microgram/mg protein, respectively). A linear relationship was also observed between nuclear zinc levels and MT immunostaining (P = 0.0024) as well as with Ki67 immunopositivity (P = 0.0123). Our results show that MT and zinc are correlated with proliferative activity in NPC, providing further insights into the biology of this enigmatic and aggressive tumour. PMID- 11023538 TI - NAD(P)H:quinone oxidoreductase-dependent risk for colorectal cancer and its association with the presence of K-ras mutations in tumors. AB - NAD(P)H:quinone oxidoreductase (NQO1) is a polymorphic enzyme involved in the detoxification of potentially mutagenic and carcinogenic quinones. The homozygous C609T NQO1 genotype resulting in loss of reductase activity is found in 2-20% of individuals. In the present study, the NQO1-dependent risk for sporadic colorectal cancer (CRC) was studied in 247 incident CRC cases and 296 hospital based controls recruited during 1996-1997. Four subgroups of cases were studied: (i) all CRCs; (ii) a molecular CRC subgroup (n = 117, cases with molecular tumor analyses); (iii) within the molecular subgroup those tumors with K-ras mutations in codon 12 (CRC K12); (iv) within the molecular subgroup those tumors with K-ras mutations in codon 13 (CRC K13). The C609T NQO1 genotype was found to be twice as prevalent in all CRC patients (6.8%) compared with controls (3%) and six times more common in the subset CRC K12 (20%). Multivariant analyses in the overall population of 247 cases and 296 controls showed a significant age and gender adjusted risk for CRC associated with the C609T NQO1 genotype (OR 2.9, 95% CI 1.19-6.97; P = 0.01) or with any variant genotype (the low activity allele frequency, i.e. heterozygotes plus homozygotes) (OR 1.41, 95% CI 1.02-1.92; P = 0.03). Within cases of the molecular subgroup (n = 117) the C609T NQO1 genotype was associated with the presence of K-ras codon 12 mutation (OR 6.5 95%, CI 1.39 34.9; P = 0.003). Logistic regression showed an age and gender adjusted risk for K-ras codon 12 mutant CRC associated with the C609T NQO1 genotype (OR 10.5, 95% CI 2.99-36.7; P: = 0.0002) or with any variant NQO1 genotype (OR 2.23, 95% CI 1.23-4.00; P = 0.007) compared with the control group. Genetically determined variations in NQO1 may modify the risk for CRC and these risks may be greatest for tumors containing K-ras codon 12 mutations. CRC with K-ras codon 12 mutations may represent a distinct and etiologically more homogeneous subtype of the disease, which may be associated with toxicants that are metabolized via a NQO1 dependent pathway. PMID- 11023539 TI - A new xeroderma pigmentosum group C poly(AT) insertion/deletion polymorphism. AB - We found a common biallelic polymorphism (PAT) in the xeroderma pigmentosum complementation group C (XPC) DNA repair gene consisting of an insertion of 83 bases of A and T [poly(AT)] and a 5 base deletion within intron 9. We developed a PCR assay to resolve the XPC PAT+ and PAT- alleles and found that the PAT+ allele frequency was 0.44 in 156 cancer-free donors from the Johns Hopkins School of Public Health, 0.41 in 263 cancer-free donors from the Baltimore Longitudinal Study of Aging and 0.36 in samples from 216 unselected donors from NIH. We also found a single nucleotide polymorphism in exon 15 of the XPC gene (A2920C, Lys939 ->Gln) that creates a new enzyme restriction site. This XPC exon 15 single nucleotide polymorphism occurred at a frequency of 0.38 in 98 NIH donors and is in linkage disequilibrium with the PAT locus. We developed an allele-specific complementation assay utilizing post-UV host cell reactivation to assess DNA repair capacity of polymorphic alleles. We found similar DNA repair with XPC 2920A and XPC 2920C. These common polymorphisms in the XPC DNA repair gene may be useful for molecular epidemiological studies of cancer susceptibility. PMID- 11023540 TI - Regulation of rat glutamate-cysteine ligase (gamma-glutamylcysteine synthetase) subunits by chemopreventive agents and in aflatoxin B(1)-induced preneoplasia. AB - Certain dietary constituents can protect against chemically induced carcinogenesis in rodents. A principal mechanism by which these chemopreventive compounds exert their protective effects is likely to be via induction of carcinogen detoxification. This can be mediated by conjugation with glutathione, which is synthesized by the sequential actions of glutamate-cysteine ligase (GLCL) and glutathione synthetase. We have demonstrated that dietary administration of the naturally occurring chemopreventive agents, ellagic acid, coumarin or alpha-angelicalactone caused an increase in GLCL activity of between approximately 3- and 5-fold in rat liver. Treatment with the synthetic antioxidant ethoxyquin or the classic inducer phenobarbital caused < 2-fold induction of GLCL activity in rat liver, which was not found to be significant. The increases in GLCL activity were accompanied by increases (between 2- and 4 fold) in levels of both the catalytic heavy subunit (GLCLC) and regulatory light subunit (GLCLR). No substantial induction of GLCL was observed in rat kidney. The glutathione S-transferase (GST) subunits A1, A3, A4, A5, P1 and M1 were all found to be inducible in rat liver by most of the agents. The greatest levels of induction were observed for GST P1, following treatment with coumarin (20-fold), alpha-angelicalactone (10-fold) or ellagic acid (6-fold), and GST A5, following treatment with coumarin (7-fold), alpha-angelicalactone (6-fold) and ethoxyquin (6-fold). Glutathione synthetase was induced approximately 1.5-fold by coumarin, alpha-angelicalactone, ellagic acid and ethoxyquin. The expression of glutathione related enzymes was also examined in preneoplastic lesions induced in rat liver by aflatoxin B(1). The majority of gamma-glutamyltranspeptidase (GGT)-positive preneoplastic foci contained increased levels of GLCLC relative to the surrounding tissue. This was usually found to be accompanied by an increase in GLCLR. Cells in the inner cortex of rat kidney were found to contain the highest levels of both GLCLC and GLCLR. The same cells showed the strongest staining for GGT activity. PMID- 11023541 TI - Potent preventive action of curcumin on radiation-induced initiation of mammary tumorigenesis in rats. AB - This investigation evaluated the preventive effect of curcumin on radiation induced tumor initiation in rat mammary glands. Fifty-four female rats were mated and then divided into two groups at day 11 of pregnancy. As the control group, 27 rats were fed a basal diet during the experimental period. As the experimental group, 27 rats were fed a diet containing 1% curcumin between day 11 of pregnancy and parturition (day 23 of pregnancy). All rats of both groups received whole body irradiation with 1.5 Gy gamma-rays from a (60)Co source at day 20 of pregnancy and were then implanted with a diethylstilbestrol pellet 1 month after weaning. A high incidence (70.3%) of mammary tumorigenesis was observed in the control group. The tumor incidence (18.5%) was significantly reduced in the rats fed curcumin during the initiation stage. The appearance of the first palpable tumor was delayed by 6 months in the curcumin-fed group and the average latent period until the appearance of mammary tumors was 2.5 months longer in the curcumin-fed group than in the control group. By histological examination, the proportion of adenocarcinoma (16.7%) in total tumors in the curcumin-fed rats was found to be decreased to half that (32.1%) in the control group. Compared with the control rats, the body weight of rats in the experimental group was decreased slightly by administration of the curcumin diet from day 11 of pregnancy, in spite of a similar intake of diet, but had recovered to the level of the control by the end of the experiment. At the time of irradiation, curcumin did not have any effect on organ weight or on the development and differentiation of mammary glands of pregnant rats. In addition, the serum concentrations of fatty acids, thiobarbituric acid-reactive substances and ovarian and pituitary hormones, except LH, remained at the control level. Also, no change in litter size and body weight of pups born from curcumin-fed rats indicated no toxicity of curcumin. These results suggest that curcumin does not have any side-effects and is an effective agent for chemoprevention acting at the radiation-induced initiation stage of mammary tumorigenesis. PMID- 11023542 TI - Suppression by citrus auraptene of phorbol ester-and endotoxin-induced inflammatory responses: role of attenuation of leukocyte activation. AB - Auraptene (AUR), a citrus coumarin derivative, is one of the promising chemopreventive agents against skin, tongue, esophagus and colon carcinogenesis in rodents. We reported previously that AUR suppresses superoxide anion (O(2)(-)) generation from inflammatory leukocytes in in vitro experiments. In the present study, we investigated the anti-inflammatory activities of AUR using a 12-O: tetradecanoylphorbol-13-acetate-treated mouse skin model, and compared them with those of umbelliferone (UMB), a structural analog of AUR that is virtually inactive toward O(2-) generation inhibition. Double pre-treatments of mouse skin with AUR, but not UMB, markedly suppressed edema formation, hydrogen peroxide production, leukocyte infiltration, and the rate of proliferating cell nuclear antigen-stained cells. These inhibitory effects by AUR are attributable to its selective blockade of the activation stage, as revealed by single pre-treatment experiments. In a murine macrophage line, RAW 264.7, AUR significantly attenuated the lipopolysaccharide-induced protein expression of inducible isoforms of both nitric oxide synthase and cyclooxygenase, with decreased production of nitrite anion and prostaglandin E(2), and yet suppressed the release of tumor necrosis factor-alpha. Conversely, UMB did not show any inhibitory effect. This contrasting activity profile between AUR and UMB was rationalized to be a result of their distinct differences in cellular uptake efficiencies, i.e. the geranyloxyl group in AUR was found to play an essential role in incorporation. Thus, our findings indicate that AUR is an effective agent to attenuate the biochemical responsiveness of inflammatory leukocytes, which may be essential for a greater understanding of the action mechanism that underlies its inhibition of inflammation-associated carcinogenesis. PMID- 11023543 TI - Major inter-species differences in the rates of O-sulphonation and O glucuronylation of alpha-hydroxytamoxifen in vitro: a metabolic disparity protecting human liver from the formation of tamoxifen-DNA adducts. AB - Tamoxifen is a hepatic genotoxin in rats and mice but a hepatocarcinogen only in rats. It is not associated with DNA adducts and liver tumours in patients. The proposed major pathway for its bioactivation in rats involves alpha hydroxylation, O-sulphonation and generation of a carbocation that reacts with DNA. Rat liver microsomes catalyse alpha-hydroxylation at approximately 2- and 4 fold the rate achieved by human and murine liver microsomes, respectively. O glucuronylation will deactivate alpha-hydroxytamoxifen and compete with sulphonation. Rates of O-sulphonation of alpha-hydroxytamoxifen in hepatic cytosol have been determined by a HPLC assay of substrate-dependent 3' phosphoadenosine 5'-phosphate production. The rank order of O-glucuronylation in hepatic microsomes was estimated by HPLC-mass spectrometry. The rate of sulphonation of trans-alpha-hydroxytamoxifen (25 microM) in cytosol from adult female Sprague-Dawley rats and CD1 mice was 5.3 +/- 0.8 and 3.9 +/- 0.5 pmol/min/mg protein (mean +/- SD, n = 3), respectively. In cytosol fractions from women aged 40-65 years, the rate was 1.1 +/- 0.4 pmol/min/mg protein (mean +/- SD, n = 6). The K(m) for trans-alpha-hydroxytamoxifen in rat, mouse and human cytosol was 84. 6 +/- 3.8, 81.4 +/- 4.6 and 104.3 +/- 5.6 microM (mean +/- SD, n = 3), respectively; the corresponding V:(max) values were 22.4 +/- 3.4, 17.1 +/- 3.1 and 6.3 +/- 1.9 pmol/min/mg protein. These K:(m) were similar to a value obtained by others using purified rat liver hydroxysteroid sulphotransferase a. Turnover of the cis epimer was too slow for accurate determination of rates. Sulphonation of trans-alpha-hydroxytamoxifen in human uterine cytosol was undetectable. The rank order of O-glucuronylation of trans-alpha-hydroxy- tamoxifen in liver microsomes was human > > mouse > rat. In combination, lower rates of alpha-hydroxylation and O-sulphonation and a higher rate of O glucuronylation in human liver would protect patients from the formation of tamoxifen-DNA adducts. PMID- 11023544 TI - The in vivo genetic activity profile of the monofunctional nitrogen mustard 2 chloroethylamine differs drastically from its bifunctional counterpart mechlorethamine. AB - The property of forming crosslinks within DNA is seen as the major cause of the high carcinogenic, genotoxic and anti-neoplastic potency of bifunctional nitrogen mustards. To further investigate the importance for genotoxicity of a second reactive group in a molecule, the genetic activity profiles of the bifunctional nitrogen mustard mechlorethamine (MEC) and its monofunctional counterpart 2 chloroethylamine (CEA) were compared, using several in vivo end points in Drosophila. When post-meiotic male germ cells were alkylated by CEA and then transferred to nucleotide excision repair (NER)-proficient oocytes, no more than up to 4-fold increased forward mutation frequencies were induced. With oocytes deficient for XPG (DmXPG), frequencies were enhanced up to 50 times. For MEC mutation frequencies increased up to 40 times the background, whereas only a low hypermutability was observed when DmXPG were used instead of wild-type females, indicating that nitrogen mustard-induced monoadducts, in contrast to crosslinks, are efficiently repaired by the NER system. Specific locus mutations generated in the vermilion gene by CEA under NER(-) conditions were almost exclusively base pair substitutions (93%). The high proportion of mutations at guanine positions indicates a strong contribution of N7-alkylguanine to the mutational spectrum. MEC induced 64% deletions and other DNA rearrangements in crosses of males with DmXPG females. The small portion of point mutations (36%) was further reduced to approximately 20% with NER(+) females. Inactivation of NER had no potentiating effect on clastogenic events (chromosome loss) induced by CEA, which is in sharp contrast to the strongly enhanced forward mutation frequencies measured with DmXPG females. The weak genotoxic effectiveness of CEA under NER(+) conditions is clearly due to efficient error-free repair of monoalkyl adducts. These results further support the concept that bifunctional nitrogen mustards exert their mutagenic activity through formation of DNA crosslinks and that DNA monoadducts make only a minor contribution to their genotoxic activity. PMID- 11023546 TI - Increased susceptibility to chemotherapeutic alkylating agents of mice deficient in DNA repair methyltransferase. AB - O(6)-methylguanine-DNA methyltransferase plays vital roles in preventing induction of mutations and cancer as well as cell death related to alkylating agents. Mice defective in the MGMT: gene, encoding the methyltransferase, were used to evaluate cell death-inducing and tumorigenic activities of therapeutic agents which have alkylation potential. MGMT(-/-) mice were considerably more sensitive to dacarbazine, a monofunctional triazene, than were wild-type mice, in terms of survival. When dacarbazine was administered i.p. to 6-week-old mice and survival at 30 days was enumerated, LD(50) values of MGMT(-/-) and MGMT(+/+) mice were 20 and 450 mg/kg body wt, respectively. Increased sensitivity of MGMT(-/-) mice to 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosou rea (ACNU), a bifunctional nitrosourea, was also noted. On the other hand, there was no difference in survival of MGMT(+/+) and MGMT(-/-) mice exposed to cyclophosphamide, a bifunctional nitrogen mustard. It appears that dacarbazine and ACNU produce O(6)-alkylguanine as a major toxic lesion, while cyclophosphamide yields other types of modifications in DNA which are not subjected to the action of the methyltransferase. MGMT(-/-) mice seem to be less refractory to the tumor-inducing effect of dacarbazine than are MGMT(+/+) mice. Thus, the level of O(6)-methylguanine-DNA methyltransferase activity is an important factor when determining susceptibility to drugs with the potential for alkylation. PMID- 11023545 TI - Failure to demonstrate chemoprevention by the monoterpene perillyl alcohol during early rat hepatocarcinogenesis: a cautionary note. AB - The monoterpene perillyl alcohol (PA) is being considered as a useful chemopreventive and therapeutic agent against human cancers. However, no data are available on the effects of PA in the first stages of hepatocarcinogenesis. To study such effects, putatively initiated cells and preneoplastic foci in hepatocarcinogenesis were used as a model. Male Wistar rats were treated with a single dose of N:-nitrosomorpholine (NNM). Between days 4 and 91 after NNM, subgroups of rats received either PA (1 g/kg body wt/day) or phenobarbital (PB) (50 mg/kg body wt/day) in the diet. Since PA treatment reduced food intake, one control group was fed ad libitum, while a second control was pair fed between days 4 and 91. In order to enhance any treatment effects, all groups, including the controls, were treated with the potent tumor promoter PB after day 91 until the end of the experiment at day 266. Rats were killed at multiple time points and putatively initiated cells and preneoplastic foci were identified by staining positively for placental glutathione S-transferase (G+). The following results were obtained. (i) A few days after NNM treatment single G+ cells emerged; a considerable portion of which developed into foci. (ii) Treatment with PB resulted in an increase in number and size of G+ foci. (iii) PA treatment failed to reduce the number of G+ cells; it somewhat lowered rates of apoptosis in G+ foci and clearly increased their average size. (iv) Eighty-seven days of PA revealed no protective effect on day 266, but, similar to PB treatment, increased the growth of foci. In conclusion, PA exerted no detectable chemopreventive effect in the early stages of rat hepatocarcinogenesis. It rather exerted a PB like tumor promoting activity. These data argue against a recommendation of PA as a chemopreventive agent for healthy humans. PMID- 11023547 TI - Inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced NF-kappaB activation by tea polyphenols, (-)-epigallocatechin gallate and theaflavins. AB - (-)-Epigallocatechin gallate (EGCG) and theaflavins are believed to be the key active components in tea for the chemoprevention of cancer. However, the molecular mechanisms by which EGCG and theaflavins block carcinogenesis are not clear. In the JB6 mouse epidermal cell line a tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA), which causes cell transformation at high frequency, markedly induced NF-kappaB activation. We found that EGCG and theaflavins inhibited TPA-induced NF-kappaB activity in a concentration-dependent manner. These polyphenols blocked TPA-induced phosphorylation of IkappaBalpha at Ser32 in the same concentration range. Moreover, the NF-kappaB sequence-specific DNA-binding activity induced by TPA was also inhibited by these polyphenols. These results suggest that inhibition of NF-kappaB activation is also important in accounting for the anti-tumor promotion effects of EGCG and theaflavins. PMID- 11023548 TI - p53 knockout mice (-/-) are more susceptible than (+/-) or (+/+) mice to N-methyl N-nitrosourea stomach carcinogenesis. AB - Mutations of the p53 tumor suppressor gene constitute one of the most frequent molecular changes in a wide variety of human cancers. Mice deficient in p53 have recently attracted attention for their potential to identify chemical genotoxins. In this study we have investigated the susceptibility of p53 nullizygote (-/-), heterozygote (+/-) and wild-type (+/+) mice to N:-methyl-N:-nitrosourea (MNU) gastric carcinogenesis. p53 knockout mice were treated with 30 p.p.m. MNU in the drinking water 1 week on and 1 week off and killed after 5 weeks. The numbers of pepsinogen-altered pyloric glands (PAPG), putative preneoplastic lesions, were 1.8, 1.7 and 22.6 in p53 (+/+), (+/-) and (-/-) mice, respectively. In a 15 week experiment, adenomas were found in 0 of 19 (+/+) (0%), 2 of 21 (+/-) (9.5%) and 6 of 10 (-/-) (60.0%) animals. Also, one well-differentiated adenocarcinoma was observed in a p53 (-/-) mouse. After 40 weeks treatment with 120 or 30 p.p.m. MNU there was no significant difference in the incidence of gastric tumors between p53 (+/+) and (+/-) mice. However, mortality from carcinogen-induced lymphomas, leukemias and sarcomas was very much greater in the latter group. Homozygous knockout animals could not be maintained long term. PCR-single strand conformation polymorphism analysis of exons 5-8 of the p53 gene of DNA extracts from 68 gastric tumors consisting of 16 and 20 30 p.p.m. MNU-treated p53 (+/+) and (+/-) mice and 14 and 18 120 p.p.m. MNU-treated p53 (+/+) and (+/-) mice demonstrated no mutations. These results suggest that p53 may not be a direct target of MNU but rather play an important role as a gatekeeper in mouse stomach carcinogenesis induced by this direct acting agent. PMID- 11023549 TI - A simple phenolic antioxidant protocatechuic acid enhances tumor promotion and oxidative stress in female ICR mouse skin: dose-and timing-dependent enhancement and involvement of bioactivation by tyrosinase. AB - The modifying effects of topical application of the phenolic antioxidant protocatechuic acid (PA) on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse skin tumor promotion were investigated. Dimethylbenz[a]anthracene-initiated female ICR mice were treated with TPA (1.6 nmol) twice weekly for 20 weeks to promote papilloma formation. Pre-treatment with 16nmol PA 30 min prior to each TPA treatment significantly inhibited the number of papillomas per mouse by 52% (P < 0.05). On the other hand, PA pre-treatment at a high dose (1600 nmol) significantly enhanced tumor numbers by 38% (P < 0.05). Interestingly, in the group treated with a quite high dose (20000 nmol) of PA 5 min prior to each TPA application, the average number of tumors per mouse was reduced by 38%, whereas the same PA dose 3 h before TPA treatment significantly enhanced tumor numbers by 84% (P < 0.01). These results suggested that topically applied PA was converted to compound(s) lacking antioxidative properties and/or rather possessing the potential to enhance tumor development. A similar tendency was also observed in the short-term experiment of TPA-induced inflammation and oxidative stress; i.e. two groups pre-treated with PA at 20000 nmol, 30min and 3h before TPA treatment, did not show suppression or even significantly enhanced TPA-induced leukocyte infiltration, H(2)O(2) generation, thiobarbituric acid-reacting substances level and proliferating cell nuclear antigen index, while PA treatment together with TPA significantly suppressed these parameters. Treatment with a high dose (20000 nmol) of PA alone for 3h enhanced oxidative stress by reducing glutathione levels in mouse skin, which was counteracted by the tyrosinase inhibitor arbutin. Oxidative stress responses such as leukocyte infiltration and H(2)O(2) generation were also counteracted by arbutin. These results suggested that tyrosinase dependent oxidative metabolism of PA was at least partially involved in the enhanced effects of PA on TPA-induced inflammatory responses and thus tumor promotion. PMID- 11023550 TI - Red meat and colon cancer: dietary haem, but not fat, has cytotoxic and hyperproliferative effects on rat colonic epithelium. AB - High intake of red meat is associated with an increased risk of colon cancer. It has been suggested that fat from red meat is responsible, because high fat intake increases the concentration of cytotoxic lipids in the colon. Experimental studies have not unequivocally supported such a role for fat, however. Recently, we showed that dietary haem, which is abundant in red meat, increased colonic cytotoxicity and epithelial proliferation. In this study, we wanted to clarify whether dietary fat affects colon cancer risk by itself or by modulating the detrimental effects of haem on the colonic epithelium. Rats were fed control or haem-supplemented diets with 10%, 25% or 40% of the energy derived from fat for 14 days. Faeces were collected for biochemical analyses. Colonic cytotoxicity was determined from the degree of lysis of erythrocytes by faecal water. Colonic epithelial proliferation was measured in vivo using [(3)H]thymidine incorporation. Increasing the fat content of the control diets stimulated faecal disposal of both fatty acids and bile acids. It also increased the concentration of fatty acids, but not that of bile acids, in faecal water in control rats. The cytolytic activity of faecal water and colonic epithelial proliferation were unaffected. Dietary haem increased faecal cation content and cytolytic activity of faecal water at all fat levels, suggesting that the colonic mucosa was exposed to high amounts of luminal irritants. This effect was smaller in rats on the low fat diet. Dietary haem also increased colonic epithelial proliferation at all fat levels. The haem-induced effects were independent of fatty acids or bile acids in the faecal water. In western societies, 30-40% of ingested energy is supplied by dietary fat, so our results suggest that the association between consumption of red meat and risk of colon cancer is mainly due to its haem content, and is largely independent of dietary fat content. PMID- 11023551 TI - ras gene mutations are absent in NMU-induced mammary carcinomas from aging rats. AB - Carcinoma induction in the rat mammary carcinogenesis model is age dependent. In this study, mammary cancer susceptibility and ras gene activation were investigated in rats exposed to N:-methyl-N:-nitrosourea (NMU) at 2, 6, 8 and 15 months. Animals were resistant to NMU-induced mammary tumor development when exposed at 6 and 8 months of age, whereas a significant number of mammary carcinomas developed in animals exposed to NMU at 2 and 15 months of age. G35- >A35 activating mutations in the Harvey ras gene were found only in mammary carcinomas from rats exposed to NMU at 2 months of age, but not in tumors that developed in animals exposed to NMU at 15 months of age. No G35-->A35 activating mutations were present in the Kirsten ras gene of any of the mammary tumors. Additional analysis of exons 1 and 2 of the Harvey ras gene from mammary carcinomas that developed in animals exposed to NMU at 15 months of age did not reveal any other activating mutations in this gene. In mammary carcinomas from animals exposed to NMU at 2 months of age, the frequency of mammary carcinomas with mutations in the Harvey ras gene was independent of the time from which the tumor first appeared. Therefore, age at the time of carcinogen exposure plays a critical role in both breast cancer susceptibility and the molecular events that contribute to mammary carcinoma development. PMID- 11023552 TI - Retinal VEGF mRNA measured by SYBR green I fluorescence: A versatile approach to quantitative PCR. AB - PURPOSE: To determine whether continuous monitoring of SYBR Green I fluorescence provides a reliable and flexible method of quantitative RT-PCR. Our aims were (i) to test whether SYBR Green I analysis could quantify a wide range of known VEGF template concentrations, (ii) to apply this method in an experimental model, and (iii) to determine whether 20 existing primer pairs could be used to quantify their cognate mRNAs. METHODS: Real-time quantitative PCR was performed using a LightCycler rapid thermal cycler (Roche). Retinal VEGF mRNA levels were measured in a murine model of oxygen-induced retinopathy during vaso-obliterative and hypoxic phases. RESULTS: This technique was able to detect as few as 10 control template copies, with quantitative data available routinely for 1000 or more copies. The levels of retinal VEGF mRNA expression followed the hypoxia-induced pattern determined previously by conventional methods. All gene-specific primer pairs which amplify a specific product by conventional PCR were successfully converted to SYBR Green analysis, including those for housekeeping genes glyceraldehyde phosphate dehydrogenase (GAPDH), cyclophilin, and acidic ribosomal phosphoprotein PO (ARP/36B4) and for 28S rRNA. In each case melting curve analysis and agarose gel electrophoresis confirmed the specificity of the amplification product. CONCLUSIONS: The sequence-independent detection of DNA with SYBR Green I means that it can be used to quantify the amplification of any cDNA using gene-specific primers. This rapid and flexible method is ideally suited for researchers in vision science wishing to quantify mRNAs from many different genes because it does not require investment in gene-specific hybridization probes. PMID- 11023553 TI - Infliximab in the treatment of Crohn's disease. PMID- 11023554 TI - Management of perianal Crohn's disease. AB - Perianal inflammation is a disabling manifestation of Crohn's disease. The primary lesions found in perianal Crohn's disease evolve in parallel with the disease elsewhere in the bowel. Although the spontaneous resolution of anal lesions is observed in up to half of patients, the penetrating nature of the disease may lead to secondary lesions including complex fistulae. In some patients, this, in turn, results in the gradual destruction of the sphincter apparatus and anal incontinence. These patients, after years of suffering, often require proctectomy. Control of activity, overall, is the first step in the management of perianal Crohn's disease. Sepsis should be controlled by the drainage of abscesses and by long term use of setons. Although antibiotics and standard immunosuppression often improve perianal fistulae, their action is usually slow and incomplete. Management of perianal Crohn's disease has changed thoroughly in the past two years since the introduction of monoclonal antibodies to tumour necrosis factor (infliximab). Complete arrest of the drainage of fistulae was obtained in 46% of patients after the administration of 5 to 10 mg/kg of infliximab at weeks 0, 2 and 6, with a median duration of effect of 12 weeks. In these patients, long term management of their bowel disease will likely require the repeated use of infliximab. Studies to evaluate this are underway. PMID- 11023555 TI - Biology of inflammation in Crohn's disease: mechanisms of action of anti-TNF-a therapy. AB - Several recent trials of intravenously administered antitumour necrosis factor alpha (TNF-a) monoclonal antibody have shown dramatic responses among patients with Crohn's disease. These results indicate a primary role for TNF-a in the mediation of altered mucosal immune function in this disease. Clinical responses in patients treated with a single infusion of anti-TNF-a persisted for as long as one year. The prolonged period of clinical benefit shows that the effect of short term TNF-a elimination remains long after the monoclonal antibody has cleared the body. Corresponding in vitro investigation has shown that T helper 1 (Th1) mediated cytokine production of interferon-gamma is downregulated in the involved mucosa to a level consistent with that seen in uninflamed mucosa. These results suggest that TNF-a-specific augmentation of mucosal Th1 function is the process that is altered by removal of TNF-a and that produces such persistent responses. Understanding how TNF-a modulates mucosal Th1 function may lead to the definition of a key feature of Crohn's disease pathogenesis. PMID- 11023556 TI - Steroid-dependent Crohn's disease. AB - Placebo controlled trials have demonstrated that a tapering course of corticosteroids is an effective therapy for active Crohn's disease. A population based study of 109 patients with Crohn's disease undergoing their first course of corticosteroids showed that, at the end of one year, 44% of patients were steroid responsive, 36% were steroid dependent and 20% were steroid refractory. Side effects occur frequently during a four-month tapering course of corticosteroids, including moon face, acne, infection, ecchymoses, hypertension, hirsutism, petechial bleeding and striae. More serious side effects occur with long term use, including hypertension, diabetes, infection, osteonecrosis, osteoporosis, myopathy, cataracts, glaucoma and psychosis. Low dose corticosteroids, alternate day corticosteroids and mesalamine (5-aminosalicylate) are not effective steroid sparing agents in patients with Crohn's disease. Controlled ileal release budesonide, 6 mg/day, is an effective steroid-sparing agent, but it does result in some decrease in adrenal function. Azathioprine, 6-mercaptopurine and methotrexate are all effective steroid-sparing agents, as is the humanized, anti tumour necrosis factor monoclonal antibody, CDP571. A preliminary, uncontrolled study has suggested that the mouse/human chimeric monoclonal antibody infliximab may also be steroid sparing. Surgical resection is an effective strategy to reduce steroid use in the short to intermediate term, but postoperative reoccurrence of Crohn's disease occurs frequently. Given the morbidity associated with prolonged corticosteroid use, medical and surgical treatment strategies to reduce steroid use should be employed routinely. PMID- 11023557 TI - Maintenance therapy in Crohn's disease. AB - Symptoms of active Crohn's disease may respond to one or more of a number of classes of drug therapies. These include systemic glucocorticoids, budesonide, sulphasalazine, mesalazine (5-aminosalicylates), immunosuppressive agents and antibiotics. More recently, a chimeric mouse-human antibody to tumour necrosis factor (infliximab) has been shown to induce clinical remission and endoscopic improvement in patients with moderately active Crohn's disease refractory to other therapies. Despite this wide range of existing therapies and the potential of emerging biological therapies, recurrent Crohn's disease continues to be a major impediment to the fulfilment of a normal lifestyle and optimal quality of life for patients with Crohn's disease. Many drugs known to be effective for the treatment of active disease have been tried as maintenance therapy to prevent disease relapse or recurrence following medical or surgical therapy. The available evidence suggests that most of these drugs are not as useful in maintaining remission as they are in inducing it. Systemic glucocorticoids, budesonide, mesalazine (5-aminosalicylates), sulphasalazine and antibiotics are all associated with either marginal therapeutic gain in the setting of maintenance therapy or unacceptable long term toxicity. The immunosuppressive agents azathioprine, 6-mercaptopurine and methotrexate have been shown to have a beneficial effect in maintaining remission and may be helpful as steroid-sparing agents. Repeated infusions of antitumour necrosis factor antibody maintain the improvements observed after one or two initial infusions. The relative long term safety, efficacy and cost effectiveness of the various choices of maintenance therapy remain to be determined. PMID- 11023558 TI - Long term safety of infliximab. AB - Infliximab is a chimeric anti-tumour necrosis factor-alpha monoclonal antibody that has been studied for the treatment of Crohn's disease and rheumatoid arthritis. In several placebo controlled, randomized clinical trials and open trials, 771 patients have been given infliximab (a further 192 received placebo). Follow-up for safety has included the time of study (12 weeks after the last infusion), plus three additional years. Acute infusion reactions (headache, fever, chills, urticaria, chest pain) were seen in 17% of patients receiving infliximab compared with 7% of those receiving placebo. While infections were reported more frequently overall in the patients given infliximab (26% over 27 weeks of follow-up versus 16% of placebo-treated patients over 20 weeks of follow up), there was no increased risk of serious infections. There was no difference in the overall mortality rate between the groups. While low titres of autoantibodies developed in less than 10% of patients, drug-induced lupus was seen in less than 1%, with these cases resolving upon discontinuation of the drug. Overall, infliximab showed an acceptable safety profile. PMID- 11023559 TI - Medical therapy of steroid-resistant Crohn's disease. AB - Glucocorticosteroids are commonly used in the treatment of moderate to severe Crohn's disease and are effective for the short term amelioration of symptoms. However, not all patients respond to corticosteroid therapy. Approximately 20% of patients fail to respond to initial treatment with steroids, while 36% of patients may be considered steroid dependent. Sharp distinctions between steroid dependent and steroid-resistant populations are difficult to draw because steroid responsiveness is often dose dependent. Clearer distinctions may be drawn between those who fail to respond to initial treatment with corticosteroids and those who have secondary failure later in the course of their disease on repeated treatment. Effective therapies for steroid-resistant Crohn's disease include azathioprine and 6-mercaptopurine, methotrexate, cyclosporine and infliximab. Limited data are available on the use of tacrolimus, mycophenolate mofetil and thalidomide. Steroid-resistant Crohn's disease, while involving a small minority of patients, continues to present a difficult therapeutic challenge. PMID- 11023560 TI - EUS in cirrhotic patients with and without prior variceal hemorrhage in comparison with noncirrhotic control subjects. AB - BACKGROUND: Endoscopic ultrasound (EUS) was used to evaluate cirrhotic patients with and without prior variceal hemorrhage. The findings were compared with those of EUS in noncirrhotic control subjects to determine EUS features indicative of cirrhosis and of a risk for variceal hemorrhage. METHODS: Patients with cirrhosis undergoing indicated endoscopic screening for varices or surveillance after endoscopic therapy for variceal hemorrhage were studied and compared with healthy noncirrhotic control patients undergoing EUS for benign conditions. RESULTS: Sixty-six cirrhotic patients (31 with prior hemorrhage) and 32 control patients were studied. Nonhemorrhage cirrhotic patients had more severe liver disease by Child's class (p = 0.02) and less beta-adrenergic blocker usage (p < 0.0001). Paraesophageal varices were detected in 97% of cirrhotic patients and 3% of control patients (p < 0.001) and were a more sensitive predictor of cirrhosis than varices at endoscopy (74%, p < 0.0001). Azygos vein and thoracic duct diameters, and gastric mucosa and submucosa thickness were greater for cirrhotic than control patients (p < 0.001). More hemorrhage patients had large (5 mm or greater) paraesophageal varices (odds ratio 3.1: 95% CI [1.1, 8.3]; p < 0.05) and paragastric varices (odds ratio 3.7: 95% CI [1.3, 10.2]; p = 0.01). Paraesophageal varix size correlated with ascites (p = 0.03) and, for nonhemorrhage patients, with Child's class (p < 0.01). CONCLUSIONS: Paraesophageal and paragastric varices correlate with the presence and severity of liver disease and portal hypertension. These data support the hypothesis that large paraesophageal and paragastric varices (5 mm or greater) may be risk factors for variceal hemorrhage, an observation that merits further prospective study. Cirrhosis causes dilation of the azygos vein and thoracic duct and thickening of gastric mucosa and submucosa. PMID- 11023561 TI - EUS in preoperative staging of pancreatic cancer. AB - BACKGROUND: Endoscopic ultrasound (EUS) is believed to be highly accurate in the local (T) and nodal (N) staging of pancreatic cancer. However, there are scant data concerning the predictive value of EUS for resectability of pancreatic adenocarcinoma. This study was performed to determine the accuracy of TNM staging by EUS in patients with pancreatic adenocarcinoma and to evaluate the role of preoperative TNM staging by EUS for determining resectability in patients with pancreatic adenocarcinoma. METHODS: This is a retrospective review of a cohort of 89 patients evaluated preoperatively with EUS for pancreatic adenocarcinoma between January 1995 and December 1997. Preoperative TNM classification by EUS was compared with surgical and histopathologic TNM staging. Resectability rates were determined and compared with the preoperative TNM staging by EUS. RESULTS: The overall accuracy of EUS for T and N staging was found to be 69% and 54%, respectively. The overall proportion of tumors that were deemed resectable by EUS and were actually found to be resectable during surgical exploration was 46%. The proportion of tumors staged as T4 N1, T4 N0, T3 N1 and T3 N0 by EUS that were found to be resectable during surgical exploration was 45%, 37%, 44% and 62%, respectively. CONCLUSIONS: In a tertiary referral patient population, EUS is not as accurate as previously reported in the T and N staging of pancreatic cancer. EUS is also not predictive of resectability in stage T3 or T4 pancreatic cancer. PMID- 11023562 TI - Endoscopic ultrasound criteria for vascular invasion in the staging of cancer of the head of the pancreas: a blind reevaluation of videotapes. AB - BACKGROUND: It has been claimed in several prospective studies that endoscopic ultrasonography (EUS) is highly accurate in the locoregional staging of pancreatic cancer. However, the value of the EUS criteria for the diagnosis of vascular involvement is less well established. To totally exclude potential bias introduced by the availability of prior information, a completely blinded analysis of videotapes of patients with cancer of the pancreatic head was therefore conducted. METHODS: Videotape sequences of 75 patients with cancer of the head of the pancreas with surgical confirmation or unequivocally positive angiography demonstrating vascular invasion were reevaluated without any clinical data or information from other imaging studies. Involvement of the vascular system (portal vein with confluence, superior mesenteric vein, celiac axis) was assessed on EUS with special emphasis on EUS parameters of the tumor-vessel relationship. RESULTS: The overall sensitivity and specificity of EUS in the diagnosis of venous invasion were 43% and 91%, respectively, when using predetermined parameters (visualization of tumor in the lumen, complete obstruction, or collateral vessels). If the parameter "irregular tumor-vessel relationship" had been added to these criteria, the sensitivity would have risen to 62%, but the specificity would have fallen to 79%. The only vascular system that could be properly visualized by EUS was the portal vein/confluence area. The positive and negative predictive values for the single parameters chosen to diagnose portal venous involvement were as follows: 42% and 33% for irregular tumor-vessel relationship, 36% and 34% for visualization of tumor in the vascular lumen, 80% and 28% for complete vascular obstruction, and 88% and 18% for collateral vessels. CONCLUSIONS: In a completely blinded evaluation of the EUS diagnosis of vascular invasion by cancer of the head of the pancreas it was not possible to find suitable morphologic parameters with clinically useful sensitivity and specificity values (over 80%). PMID- 11023564 TI - Effectiveness of ERCP in cholangitis: a community-based study. AB - BACKGROUND: Although experts have demonstrated the efficacy of endoscopic retrograde cholangiopancreatography (ERCP) in cholangitis, the effectiveness of ERCP in unselected patients has not been measured. The aim was to investigate the clinical impact of ERCP performed at any time and of early ERCP (within 24 hours of admission) in patients with a primary discharge diagnosis of cholangitis. METHODS: A retrospective record review of patients admitted to eight area hospitals with an International Classification of Diseases (ICD)-9 diagnosis consistent with cholangitis was performed. Extracted data included clinical characteristics, ERCP findings, and patient outcome. The associations of ERCP overall and early ERCP with length of stay were examined. Confounding factors including severity of illness, etiology of cholangitis, and hospital type were adjusted for in a multivariate analysis. RESULTS: A total of 116 patients were studied. ERCP was performed in 71 patients with endoscopic therapy administered in 57 (80%). ERCP overall was not associated with any change in length of hospital stay. However, compared with other invasive biliary procedures, ERCP was associated with a shorter hospital stay (median 5 vs. 9.5 days, p = 0.01) and a 36% (95% CI [5%, 57%]) reduction in severity-adjusted length of stay. Patients who had early ERCP had a significantly shorter hospital stay than those who had delayed ERCP (median 4 vs. 7 days, p < 0.005) and early ERCP was associated with a 34% (95% CI [11%, 48%]) reduction in severity-adjusted length of stay. CONCLUSION: Early ERCP may be an effective strategy for shortening the length of stay in patients hospitalized with cholangitis. PMID- 11023563 TI - Success of ERCP at a referral center after a previously unsuccessful attempt. AB - BACKGROUND: The diagnostic and therapeutic success of endoscopic retrograde cholangiopancreatography (ERCP) depends on a number of factors. When an attempt at ERCP fails, the physician must decide whether to repeat the procedure, rely on another imaging procedure (noninvasive or invasive), or refer to another endoscopist/center. Our aim in this prospective study was to determine the role of a second attempt at ERCP at a referral ERCP center. METHODS: Five hundred sixty-two patients were referred for ERCP after having undergone a previous unsuccessful attempt to visualize the clinically relevant duct(s). RESULTS: The overall success in visualizing the desired duct was 96.4% (542 of 562). Advanced techniques for cannulation were used in 41% (229 of 562). Anatomic abnormalities possibly contributing to the previous lack of success were present in 27% of cases. ERCP with or without manometry identified a cause or potential cause for the signs and symptoms in 86% of patients. Sixty complications occurred in 57 patients (10.1%). ERCP was unsuccessful in 20 patients (3.6%). CONCLUSIONS: The cannulation success rate and diagnostic yield of further ERCP with an acceptable complication rate warrant consideration of referral to centers with available resources and expertise. PMID- 11023565 TI - Pigtail stents: an alternative in the treatment of difficult bile duct stones. AB - BACKGROUND: When bile duct stones cannot be removed after sphincterotomy by balloon or basket extraction, stent placement can serve as a bridge to additional procedures. Biliary stents may also fragment large stones, allowing them to pass spontaneously or making them easier to extract at a later time. METHODS: Twenty patients with difficult to extract bile duct stones were prospectively studied. The patients underwent ERCP and placement of a 7F double-pigtail stent in the bile duct for 6 months. RESULTS: In seven patients (35%), repeat ERCP revealed no stones in the duct. Four patients (20%) had small stone fragments that were easily extracted with a balloon. Six patients (30%) continued to have large stones at repeat ERCP; two of these patients eventually underwent surgery, the duct was cleared in three with mechanical lithotripsy, and one had long-term stenting. Three patients (15%) did not undergo repeat ERCP and were therefore treated with long-term stent placement. CONCLUSIONS: This study suggests that placement of a pigtail biliary stent is a safe and effective alternative in the management of bile duct stones that resist extraction if lithotripsy is not available. After 6 months of stent placement, stones may pass or become easier to remove in a significant proportion of patients. PMID- 11023566 TI - Sphincterotomy-associated biliary strictures: features and endoscopic management. AB - BACKGROUND: "Sphincterotomy stenosis" is a recognized late complication of endoscopic biliary sphincterotomy. The narrowing is limited to the biliary orifice and can be managed simply by repeat sphincterotomy. A similar but poorly characterized post-sphincterotomy complication involves narrowing that extends from the biliary orifice for a variable distance along the bile duct, beyond the duodenal wall. This lesion cannot be managed by repeating the sphincterotomy. METHODS: Six patients (3 men) are described with sphincterotomy associated biliary strictures, all smooth and high grade, presenting at a median of 19 months (range 8 to 60 months) after sphincterotomy. Further sphincterotomy was not possible as an intra-duodenal segment of bile duct was no longer visible. Endoscopic management consisted of serial incremental stent exchange at 2- to 4 month intervals. The goal of therapy was to place two 11.5F stents side-by-side. RESULTS: Stricture resolution was documented by cholangiography in all patients. One patient with a stricture resistant to treatment required three 10F stents side-by-side, and another underwent treatment to a maximum of adjacent 11.5F and 7F stents. Two 11.5F stents were eventually placed in the other four patients. Overall median duration of stent placement was 12.5 months. At a median of 26.5 months of stent-free follow-up, all patients remain asymptomatic. CONCLUSION: Sphincterotomy-associated biliary strictures are a distinct late complication of biliary sphincterotomy. These recalcitrant lesions are not amenable to repeat sphincterotomy; however, the results of this study suggest that they may be managed successfully by serial placement of stents of incrementally increasing diameter. PMID- 11023567 TI - Endoscopic therapy for stenosis of the biliary and pancreatic duct orifices. AB - BACKGROUND: Manipulation of the duodenal papilla may lead to symptomatic stenosis of the orifices of bile duct, main pancreatic duct or accessory pancreatic duct. METHODS: Seventeen patients with stenosis of the orifice (bile duct 7, bile duct/main pancreatic duct 7, accessory pancreatic duct 3) underwent sphincterotomy and/or dilation and stent placement for a median of 140 days (range 30 to 1080 days). Patients were interviewed at a median of 720 days (range 120 to 990 days) after removal of the final stent. RESULTS: Median age was 50 years (range 17 to 68 years); 78% were women. The etiology of stenosis of the orifice was sphincterotomy in 8, sphincteroplasty in 7 and papillectomy in 2 patients. Indications for treatment were abdominal pain (100%), dilated bile duct and/or main pancreatic duct (14 patients) and pancreas divisum (3 patients). Sixty procedures (median 4 per patient) were performed with mild morbidity (hospital stay less than 3 days) in 17% of procedures and 35% of patients. Symptoms improved in 100%, 57% and 33% of patients with bile duct, bile duct/main pancreatic duct and accessory pancreatic duct, respectively. Surgery was ultimately needed in 3 (43%) patients with bile duct/main pancreatic duct and 2 (67%) with accessory pancreatic duct stenosis. CONCLUSIONS: Endoscopic therapy successfully relieves pain due to biliary stenosis of the orifice but less frequently relieves pain due to pancreatic stenosis of the orifice. PMID- 11023568 TI - Transnasal versus transoral endoscopy for the placement of nasoenteral feeding tubes in critically ill patients. AB - BACKGROUND: Nasoenteral feeding tube placement with the Seldinger technique using transoral endoscopy is a tedious procedure. This study compared the transoral approach with a new technique that uses a transnasal endoscope without the need for a mouth-to-nose wire transfer. METHODS: Critically ill patients requiring nasoenteral feeding tube placement were randomly assigned to the transoral technique using a standard upper endoscope (n = 80) or the transnasal method using a 5.3 mm fiberscope (n = 80). Procedure time, medication requirement, technical difficulty, patient tolerance, and radiologic tube position were assessed. RESULTS: The two groups were similar with regard to baseline medication, endoscopic findings, as well as overall technical difficulty and patient tolerance. The transnasal technique required less procedure time (median 8.0 versus 12.0 minutes, p < 0.001) and less relaxant medication (p = 0.029). Furthermore, it caused fewer circulatory (p = 0.040) and respiratory (p = 0.016) alterations regardless of the application of sedative or relaxant medication. The transnasal endoscope was inferior with respect to passage through the pylorus (p = 0.003) and duodenum (p = 0.020). These differences were significant in univariate hypothesis testing. Bonferroni correction for multiple testing of data removed the significance at p > 0.0031. Both techniques achieved similar rates of successful tube placement in the small bowel (86% versus 84%, p = 0.82). CONCLUSION: Transnasal endoscopy allows accurate placement of nasoenteral feeding tubes in critically ill patients and is superior to transoral endoscopy in terms of procedure time, medication requirement, and safety. PMID- 11023569 TI - Mucosal abnormalities of the colon in patients with portal hypertension: an endoscopic study. AB - BACKGROUND: Controversy still exists regarding colonic mucosal abnormalities in patients with portal hypertension (portal colopathy). The aims of this study were to better define portal colopathy and to identify risk factors for these colonic mucosal abnormalities. METHODS: We reviewed the medical records of 437 patients with cirrhosis and portal hypertension and 224 with irritable bowel syndrome (control patients) who underwent colonoscopy over a 6-year period. RESULTS: Individuals with portal hypertension were significantly more likely than control patients to have colitis-like abnormalities (38% vs. 3%, p < 0.001) and vascular lesions (13% vs. 3%, p < 0.001). In the multivariate model, portal hypertensive gastropathy (odds ratio 5.64: 95% CI [3.39, 9.41]; p < 0.001), 2+ or larger esophageal varices (odds ratio 4.76: 95% CI [2. 78, 8.15]; p < 0.001), and Child Pugh class C cirrhosis (odds ratio 2.64: 95% CI [1.40, 4.97]; p = 0.003) were independently associated with an increased risk of having portal colopathy, whereas the use of beta-blockers independently decreased the risk of having these findings (odds ratio 0.23: 95% CI [0.13, 0.40]; p < 0.001). Mucosal biopsies of the colon in patients with colitis-like abnormalities revealed a mild, nonspecific inflammatory infiltrate with edema and vascular ectasias in the majority of cases. CONCLUSIONS: Mucosal abnormalities in portal colopathy include edema, erythema, granularity, friability, and vascular lesions, findings that may be confused with colitis. A standardized grading system to classify the endoscopic appearance and severity of portal colopathy should be adopted. PMID- 11023570 TI - Waiting lines in the endoscopy unit. AB - BACKGROUND: The efficient management of the endoscopy unit involves issues of scheduling. The aim of this study was to assess efficient use of an endoscopy unit and its relationship to patient waiting. METHODS: Formulas of queueing theory are applied to patient scheduling in endoscopy. The M/M/n queueing model assumes exponential (Markovian) distributions underlying both patient arrival and endoscopy times with n as the number of endoscopists, while the D/M/n model assumes a constant (deterministic) patient arrival rate. RESULTS: As the use of the facility increases, so does the probability that patients will have to wait. It is impossible for an endoscopy facility to have a 100% rate of use without patients being forced to wait. Any increase in the use rate of an endoscopy unit results in a concomitant increase in waiting times. A facility with multiple endoscopists can reduce waiting probability at a higher rate of facility use. A table is included that physicians and administrators can use as a guide to estimate the use rate and waiting characteristics of different types of endoscopy facilities. CONCLUSIONS: Endoscopy units must find a balance between patient waiting and underuse of their resources. Endoscopists will occasionally find themselves idle at the benefit of providing patients with tolerable waiting times. It needs to be explained to patients that even under the best of circumstances some waiting cannot be prevented. PMID- 11023571 TI - Traumatic cholecystitis. PMID- 11023572 TI - Mucin-hypersecreting cholangiocarcinoma. PMID- 11023573 TI - Impacted chicken bone of the sigmoid colon: endoscopic removal with balloon dilatation. PMID- 11023574 TI - Schwannoma of the sigmoid colon. PMID- 11023575 TI - Preoperative staging of rectal cancer using a 7.5 MHz front-loading US probe. AB - BACKGROUND: Conventional echoendoscopes have disadvantages when used for staging colorectal cancer including the inability to pass the instrument through tight stenosis and limited maneuverability. This study evaluated the preoperative use of a newly developed 7.5 MHz front-loading ultrasound probe (FLUP) for local staging of rectal cancer. METHODS: A 7.5 MHz FLUP, diameter 7.3 mm, was used in this study. The mechanical shaft portion of the probe can be passed in retrograde fashion through the accessory channel of a standard colonoscope. Thirty-nine patients with rectal cancer underwent ultrasonography with this probe. The tumors were staged using the TNM system, and the results were compared with the histologic findings of the resected specimens. RESULTS: The FLUP proved to be satisfactory, with respect to maneuverability, for traversing stenosis and accurate recognition of small tumors under direct endoscopic control. The accuracy of the FLUP for T staging was 82% (32 of 39) for all tumors, 90% in pT1, and 79% in pT2 to pT4 tumors. The accuracy of the FLUP for N staging was 72% (23 of 32) overall. The sensitivity was 83%, the specificity was 65%, the positive predictive value was 59%, and the negative predictive value was 87%. CONCLUSIONS: The 7.5 MHz FLUP appears to be useful for preoperative local staging of rectal cancer. This system makes it technically easier to image small cancers as well as advanced rectal cancers. PMID- 11023576 TI - EUS-guided fine-needle aspiration cytodiagnosis of hilar cholangiocarcinoma: a case series. AB - BACKGROUND: Improved methods of tissue diagnosis for obstruction at the hilum of the liver (porta hepatis) have contributed substantially to the preoperative diagnosis of cholangiocarcinoma. Endoscopic brushing during endoscopic retrograde cholangiopancreatography (ERCP), with sensitivity of 20% to 100%, is the preferred technique for obtaining accurate pathologic results. Extensive hepatic resection with curative intent as well as modern approaches to palliative treatment are based on definitive diagnosis. This is a study involving endosonography-guided, fine-needle aspiration (EUS-FNA) for cytodiagnosis of potentially operable hilar cholangiocarcinoma when brush cytology was negative or unavailable. METHODS: Ten consecutive patients (7 men, 3 women; age 47 to 78 years, median 59 years) with bile duct strictures at the hepatic hilum, diagnosed by CT and/or ERCP, underwent EUS-FNA using linear echoendoscopes and 22-gauge needles. RESULTS: Adequate material was obtained in nine patients. Cytology revealed cholangiocarcinoma in seven and hepatocellular carcinoma in one. One benign inflammatory lesion identified on cytology proved to be a false-negative finding by frozen section. Metastatic locoregional hilar lymph nodes were detected in two patients, and in one patient the celiac and para-aortic lymph nodes were aspirated to obtain tissue proof of distant metastasis. There were no complications. CONCLUSIONS: When standard methods of tissue diagnosis are inconclusive, EUS-guided FNA may have a potential role in the diagnosis of primary cholangiocarcinoma of the hepatic hilum. As a new, minimally invasive approach, it proved to be technically feasible without significant risks. PMID- 11023577 TI - Endoscopic transhepatic repair of the common hepatic duct after excision during cholecystectomy. PMID- 11023578 TI - Secondary esophageal tuberculosis in a patient with thymoma. PMID- 11023579 TI - EUS-guided transesophageal pancreatic pseudocyst drainage. PMID- 11023580 TI - A bleeding gastric ulcer on a vanishing tumor caused by anisakiasis. PMID- 11023581 TI - Peutz-Jeghers syndrome managed with a complete intraoperative endoscopy and extensive polypectomy. PMID- 11023582 TI - Heparin provocation for endoscopic localization of recurrent obscure GI hemorrhage. PMID- 11023583 TI - Hemoclip repair of a sphincterotomy-induced duodenal perforation. PMID- 11023584 TI - Rectus muscle abscess associated with endoscopic tattooing of the colon with India ink. PMID- 11023585 TI - A case of intramural esophageal hematoma: complication of anticoagulation with heparin. PMID- 11023586 TI - Esophageal web in Zinsser-Engman-Cole-Fanconi disease. PMID- 11023587 TI - Pancreatic glucagonoma as a rare cause of chronic obstructive pancreatitis. PMID- 11023588 TI - Needle knife precut-induced bleeding from a "pancreatic cyst". PMID- 11023589 TI - Gastroduodenal intussusception secondary to a giant solitary gastric heterotopia: a case report. PMID- 11023590 TI - Plastic prosthesis in the palliation of small bowel stenosis secondary to recurrent gastric cancer: initial cost savings. PMID- 11023591 TI - Peutz-Jeghers syndrome associated with adenocarcinoma and protein-losing enteropathy. PMID- 11023592 TI - The value of EUS in the management of portal hypertension. PMID- 11023593 TI - Role of EUS in the evaluation of pancreatic adenocarcinoma. PMID- 11023594 TI - The flexible sigmoidoscope. PMID- 11023595 TI - Colonoscopy versus flexible sigmoidoscopy in evaluation of chronic diarrhea. PMID- 11023596 TI - Response PMID- 11023597 TI - Measuring location of neoplasms and lesions in the rectum. PMID- 11023598 TI - Molecular cloning and developmental expression of AtGR1, a new growth-related Arabidopsis gene strongly induced by ionizing radiation. AB - Screening for mRNAs that accumulate after DNA damage induced by ionizing radiation, we have isolated a 2.0-kb cDNA coding for a new Arabidopsis PEST-box protein named AtGR1 (A. thaliana gamma response 1) with an expression profile similar to that observed for several plant cell cycle-related proteins. Using an anti-AtGR1 antibody, we have shown that the AtGR1 protein is expressed at basal levels in mitotically dividing cells (meristematic tissues and organ primordia) and at a strongly enhanced level in endoreduplicating cells (stipules, trichomes). Using transgenic Arabidopsis plants that express the GUS reporter gene under the control of the AtGR1 promoter, we have demonstrated that the observed AtGR1 protein distribution is due to the promoter activity. Our results suggest that basal AtGR1 levels are associated with progression through mitosis, whereas elevated intracellular levels of AtGR1 seem to induce changes between the S and M phases of the cell cycle that trigger somatic cells to enter the endoreduplication cycle. Ionizing radiation-induced rapid and dose-dependent accumulation of AtGR1 mRNA in cell cultures and plant tissues leads to tissue specific accumulation of AtGR1 protein, best observed in ovules, which never undergo an endoreduplication cycle. It therefore appears that the radiation induced transient AtGR1 accumulation reflects DNA damage-dependent transient cell cycle arrest before mitosis, which is necessary to accomplish DNA repair prior to chromosome segregation and cytokinesis. PMID- 11023599 TI - Decreased pulmonary radiation resistance of manganese superoxide dismutase (MnSOD)-deficient mice is corrected by human manganese superoxide dismutase Plasmid/Liposome (SOD2-PL) intratracheal gene therapy. AB - The pulmonary ionizing radiation sensitivity of C57BL/6 Sod2(+/-) mice heterozygous for MnSOD deficiency was compared to that Sod2(+/+) control littermates. Embryo fibroblast cell lines from Sod2(-/-) (neonatal lethal) or Sod2(+/-) mice produced less biochemically active MnSOD and demonstrated a significantly greater in vitro radiosensitivity. No G(2)/M-phase cell cycle arrest after 5 Gy was observed in Sod2(-/-) cells compared to the Sod2(+/-) or Sod2(+/+) lines. Subclonal Sod2(-/-) or Sod2(+/-) embryo fibroblast lines expressing the human SOD2 transgene showed increased biochemical activity of MnSOD and radioresistance. Sod2(+/-) mice receiving 18 Gy whole-lung irradiation died sooner and had an increased percentage of lung with organizing alveolitis between 100 and 160 days compared to Sod2(+/+) wild-type littermates. Both Sod2(+/-) and Sod2(+/+) littermates injected intratracheally with human manganese superoxide dismutase-plasmid/liposome (SOD2-PL) complex 24 h prior to whole-lung irradiation showed decreased DNA strand breaks and improved survival with decreased organizing alveolitis. Thus underexpression of MnSOD in the lungs of heterozygous Sod2(+/-) knockout mice is associated with increased pulmonary radiation sensitivity and parallels increased radiation sensitivity of embryo fibroblast cell lines in vitro. The restoration of cellular radioresistance in vitro and in lungs in vivo by SOD2-PL transgene expression supports a potential role for SOD2-PL gene therapy in organ-specific radioprotection. PMID- 11023600 TI - In vitro and in vivo expression of endothelial von Willebrand factor and leukocyte accumulation after fractionated irradiation. AB - Previous investigations have demonstrated an increased release of von Willebrand factor (VWF; also known as vWF) in endothelial cells after high single-dose irradiation in vitro. We have also found increased levels of Vwf protein in mouse glomeruli after a high single dose of renal irradiation in vivo. In addition, increased numbers of leukocytes were observed in the renal cortex after irradiation in vivo. The aim of the present study was to investigate and quantify these biological processes after clinically relevant fractionated irradiation and to relate them to changes in renal function. A significantly greater increase in release of VWF was observed in cultured human umbilical vein endothelial cells (HUVECs) after fractionated irradiation (20 x 1.0 Gy) than after a single dose of 20 Gy (147% compared to 115% of control, respectively, P < 0.0005). In contrast with the in vitro observations, glomerular Vwf staining was lower after fractionated irradiation in vivo (20 x 2.0 Gy or 10 x 1.6 Gy +/- re-irradiation) than after a single dose of 16 Gy. The number of leukocytes accumulating in the renal cortex was also lower after fractionated in vivo irradiation than after a single radiation dose. The onset of these events preceded renal functional and histopathological changes by approximately 10 weeks. These data indicate that radiation-induced changes in endothelial VWF expression after in vivo irradiation may be distinct from the in vitro observations. Increased VWF expression may reflect pivotal processes in the pathogenesis of late radiation nephropathy and provide a clue to appropriate timing of pharmacological intervention. PMID- 11023601 TI - Comparative risk assessment of secondary cancer incidence after treatment of Hodgkin's disease with photon and proton radiation. AB - Probabilities for secondary cancer incidence have been estimated for a patient with Hodgkin's disease for whom treatment has been planned with different radiation modalities using photons and protons. The ICRP calculation scheme has been used to calculate cancer incidence from dose distributions. For this purpose, target volumes as well as critical structures have been outlined in the CT set of a patient with Hodgkin's disease. Dose distributions have been calculated using conventional as well as intensity-modulated treatment techniques using photon and proton radiation. The cancer incidence has been derived from the mean doses for each organ. The results of this work are: (a) Intensity-modulated treatment of Hodgkin's disease using nine photon fields (15 MV) results in nearly the same cancer incidence as treating with two opposed photon fields (6 MV). (b) Intensity-modulated treatment using nine proton fields (maximum energy 177.25 MeV) results in nearly the same cancer incidence as treating with one proton field (160 MeV). (c) Irradiation with protons using the spot scanning technique decreases the avoidable cancer incidence compared to photon treatment by a factor of about two. This result is independent of the number of beams used. Our work suggests that there are radiotherapy indications in which intensity-modulated treatments will result in little or no reduction of cancer incidence compared to conventional treatments. However, proton treatment can result in a lower cancer incidence than photon treatment. PMID- 11023602 TI - Quantitative image analysis of laminin immunoreactivity in skin basement membrane irradiated with 1 GeV/nucleon iron particles. AB - We previously reported that laminin immunoreactivity in mouse mammary epithelium is altered shortly after whole-body irradiation with 0.8 Gy from 600 MeV/nucleon iron ions but is unaffected after exposure to sparsely ionizing radiation. This observation led us to propose that the effect could be due to protein damage from the high ionization density of the ion tracks. If so, we predicted that it would be evident soon after radiation exposure in basement membranes of other tissues and would depend on ion fluence. To test this hypothesis, we used immunofluorescence, confocal laser scanning microscopy, and image segmentation techniques to quantify changes in the basement membrane of mouse skin epidermis. At 1 h after exposure to 1 GeV/nucleon iron ions with doses from 0.03 to 1.6 Gy, neither the visual appearance nor the mean pixel intensity of laminin in the basement membrane of mouse dorsal skin epidermis was altered compared to sham irradiated tissue. This result does not support the hypothesis that particle traversal directly affects laminin protein integrity. However, the mean pixel intensity of laminin immunoreactivity was significantly decreased in epidermal basement membrane at 48 and 96 h after exposure to 0.8 Gy 1 GeV/nucleon iron ions. We confirmed this effect with two additional antibodies raised against affinity-purified laminin 1 and the E3 fragment of the long-arm of laminin 1. In contrast, collagen type IV, another component of the basement membrane, was unaffected. Our studies demonstrate quantitatively that densely ionizing radiation elicits changes in skin microenvironments distinct from those induced by sparsely ionizing radiation. Such effects may might contribute to the carcinogenic potential of densely ionizing radiation by altering cellular signaling cascades mediated by cell-extracellular matrix interactions. PMID- 11023603 TI - Immediate localized CDKN1A (p21) radiation response after damage produced by heavy-ion tracks. AB - Using confocal microscopy on immunofluorescence-stained cells, we have investigated the response of CDKN1A (p21), one of the key proteins involved in the DNA damage response pathway, after irradiation with accelerated lead or chromium ions. Each traversal of an accelerated ion leads to the formation of a single, bright focus of the CDKN1A protein in the nuclei of human fibroblasts within 2 min after irradiation at 4 degrees C. This immediate, localized CDKN1A response is specific for particle irradiation with a high linear energy transfer (LET), whereas X irradiation, after a period of induction, yields a diffusely spread pattern, in line with the differences in the microscopic dose deposition pattern of both radiation types. The particle-induced CDKN1A foci persist for several hours until they become diffuse and vanish. These findings suggest that CDKN1A accumulates at the sites of primary DNA damage, possibly mediated by the interaction with proteins involved in DNA repair. Here, for the first time, an immediate biological response confined to the radial extension of low-energy particle tracks ( approximately 1 micrometer) is directly visualized and correlated to ion traversals. This indicates that particle irradiation represents an ideal tool to study the processing of biological damage induced in defined subnuclear regions. PMID- 11023604 TI - Effects of low-dose neutrons applied at reduced dose rate on human melanoma cells. AB - Human melanoma cells that are resistant to gamma rays were irradiated with 14 MeV neutrons given at low doses ranging from 5 cGy to 1.12 Gy at a very low dose rate of 0.8 mGy min(-1) or a moderate dose rate of 40 mGy min(-1). The biological effects of neutrons were studied by two different methods: a cell survival assay after a 14-day incubation and an analysis of chromosomal aberrations in metaphases collected 20 h after irradiation. Unusual features of the survival curve at very low dose rate were a marked increase in cell killing at 5 cGy followed by a plateau for survival from 10 to 32.5 cGy. The levels of induced chromosomal aberrations showed a similar increase for both dose rates at 7.5 cGy and the existence of a plateau at the very low dose rate from 15 to 30 cGy. The existence of a plateau suggests that a repair process after low-dose neutrons might be induced after a threshold dose of 5-7.5 cGy which compensates for induced damage from doses as high as 32.5 cGy. These findings may be of interest for understanding the relative biological effectiveness of neutrons and the effects of environmental low-dose irradiation. PMID- 11023605 TI - Neutron RBE for induction of tumors with high lethality in Sprague-Dawley rats. AB - The effectiveness of fission neutrons is compared to that of gamma rays and X rays with regard to the induction of malignancies in male Sprague-Dawley rats. The analysis is based on autopsy results. It is focused on tumors that tend to be present in animals dying early, which is indicative of a high degree of lethality. The relative biological effectiveness (RBE) is deduced from a comparison of the cumulative hazard functions. Different nonparametric models-the constant relative risk model, a time shift model, and an acceleration model-are employed in the comparison, and the resulting values of RBE are seen to be substantially independent of the choice of model. The results are in good agreement with earlier studies of nonlethal lung tumors in the same series of experiments. At neutron doses of 20 to 60 mGy, the RBE of fission neutrons is about 50. PMID- 11023606 TI - Hsp25-induced radioresistance is associated with reduction of death by apoptosis: involvement of Bcl2 and the cell cycle. AB - We previously demonstrated the protective effect of the small heat-shock protein against oxidative damage induced by tumor necrosis factor alpha. Here we have extended our studies of the possible role of Hsp25 in ionizing radiation-induced damage. For these studies, we transfected murine fibroblast L929 cells with the Hsp25 gene and selected three stably transfected clones. Hsp25 overexpression conferred radioresistance as detected by clonogenic survival and induction of apoptosis. Interestingly, the Hsp25-transfected cells showed an increase in the level of the anti-apoptosis molecule Bcl2. We also observed alterations of cell growth in the Hsp25-transfected cells. The cell cycle time of Hsp25-transfected cells was 3-4 h slower than that of vector-transfected control cells. Flow cytometry analysis of synchronized cells at late G(1) phase by mimosine treatment also showed the growth delay in Hsp25-overexpressing cells. In addition, reduced cyclin D1, cyclin A and Cdc2 levels and increased levels of Cdkn1a (also known as p21(Waf)) were observed in Hsp25-transfected cells, which probably caused the reduction in cell growth. In addition, synchronization by mimosine treatment only partially altered radioresistance in the Hsp25-transfected cells. Taken together, these data suggest that Hsp25-induced radioresistance is associated with growth delay as well as induction of Bcl2. PMID- 11023607 TI - Targeting apoptosis by hydroxymethylacylfulvene in combination with gamma radiation in prostate tumor cells. AB - Hydroxymethylacylfulvene (HMAF) is a novel agent with alkylating activity and is a potent inducer of apoptosis that is currently undergoing Phase II clinical trials for prostate cancer. This study explored the pro-apoptosis and anti proliferative potential of HMAF in combination with gamma radiation in human prostate tumor cell lines. Apoptosis was assessed based on the generation of fragmented DNA, a terminal transferase flow cytometry assay, and cell morphology. In each of the tumor cell lines examined, radiation alone induced a marginal level of apoptosis, even after a prolonged 48-h incubation after exposure. In contrast, HMAF alone was a potent inducer of apoptosis in prostate tumor cells but not in normal cells. Marked levels of apoptosis in tumor cells were also observed for the combination of HMAF with gamma radiation. When drug treatment preceded irradiation, at least additive levels of apoptosis were observed in both androgen-responsive and androgen-independent cells. The combined treatment with ionizing radiation and HMAF reduced the radiation dose needed for the same level of clonogenic survival up to 2.5-fold. The potentiation of apoptosis and reduction in the clonogenic survival of tumor cells occurred at HMAF concentrations lower than that which reduced survival to 10% and at doses up to 6 Gy. No potentiation of apoptosis or clonogenic inhibition was noted in normal cells. These results suggest that the combination of HMAF with gamma radiation may have clinical utility for treatments of prostate cancer. PMID- 11023608 TI - Measurement of hypoxia using the comet assay correlates with preirradiation microelectrode pO2 histography in R3327-AT rodent tumors. AB - Polarographic determination of tumor oxygenation by Eppendorf histography is currently under investigation as a possible predictor of radiotherapy outcome. Alternatively, the alkaline comet assay has been proposed as a radiobiological approach for the detection of hypoxia in clinical tumor samples. Direct comparisons of these methods are scarce. One earlier study with different murine tumors could not establish a correlation, whereas a weak correlation was reported for a variety of human tumors. Considering the different end points and spatial resolution of the two methods, a direct comparison for a single tumor entity appeared desirable. Anaplastic R3327-AT Dunning prostate tumors were grown on Copenhagen rats to volumes of 1-6 cm(3). Eppendorf histography (100-200 readings in 5 parallel tracks) for 8 different tumors revealed various degrees of oxygenation, with median pO(2) values ranging from 1.1 to 23 mmHg. Within 5 min after an acute exposure to 8 Gy (60)Co gamma rays, tumors were excised from killed animals and rapidly cooled to limit repair, and a single cell suspension was prepared for use with the comet assay. The resulting comet moment distributions did not exhibit two subpopulations (one hypoxic and the other aerobic), and a hypoxic fraction could not be calculated. Instead, the average comet moment distribution was taken as a parameter of overall strand break induction. Corresponding experiments with tumor cells grown in vitro allowed us to derive the relationship between the oxygen enhancement ratio (OER) for the average comet moment and oxygen partial pressure (Howard-Flanders and Alper formula). The validity of this relationship was inferred for cells exposed in situ, and the convolution of a pO(2) distribution with the formula of Howard Flanders and Alper yielded an array of expected OER values for each tumor. The average expected OER correlated well with the average comet moment (r = 0.89, P < 0.01), and the in situ comet moment distributions could be predicted from the Eppendorf data when 50% repair was taken into account, assuming a 5-min damage half-life. The findings confirm the potential of interstitial polarography to reflect radiobiologically relevant intracellular oxygenation, but also underscore the confounding influence of differences in repair that may occur when cells are prepared from irradiated tissues for use with the comet assay. PMID- 11023609 TI - Strain differences in carcinogenic and hematopoietic responses of mice after injection of plutonium citrate. AB - The carcinogenicity of injected (239)Pu citrate was compared in female mice of the C3H, C57BL/6 and BC3F(1) hybrid strains with different spectra of spontaneous or radiation-induced tumors. A significant reduction in survival due to early death caused particularly by the induction of osteosarcomas was noted in each strain after injection of 500 Bq or more. The dose response of osteosarcomas appeared to have a similar pattern in each strain except for the differences in the skeletal dose ranges for the maximum induction. While the incidence of lymphoid tumors decreased as that of osteosarcomas increased sharply to the maximum at higher doses, their histological phenotypes were predominantly non thymic, pre-B-cell leukemic lymphomas compared to the controls in each strain. Myeloid leukemias were not highly induced in any of the control and (239)Pu injected mice, and solid tumors involving the other organs were reduced in each strain after injection of 500 Bq or more. To follow up the hematological kinetics related to alpha-particle irradiation of bone marrow stem cells, sequential examinations were done in mice of each strain within 1 year after injection of 5000 Bq. The numbers of peripheral white blood cells and bone marrow cells were consistently reduced in each strain from 90 days on, while spleen cells increased from 180 days on. Granulocyte-macrophage and macrophage colony-forming cells were also consistently reduced in the bone marrow, with a compensatory increase in the spleen from 90 days on. These findings indicate that the carcinogenic and hematopoietic responses were specific to alpha-particle irradiation and were independent of mouse strain after injection with (239)Pu citrate. PMID- 11023611 TI - Effects of modifying topoisomerase II levels on cellular recovery from radiation damage. AB - Effects of Modifying Topoisomerase II Levels on Cellular Recovery from Radiation Damage. Experiments were performed with the budding yeast, Saccharomyces cerevisiae, to test whether DNA topoisomerase II is involved in repair of DNA damage induced by ionizing radiation. Topoisomerase II was inactivated by use of a temperature-sensitive mutation. Enzyme inactivation increased cellular radiosensitivity, blocked the restitution of broken chromosomes, assayed by pulsed-field gel electrophoresis, and prolonged the induction of a DNA damage inducible gene (RNR3). Overexpression of the topoisomerase II gene did not alter cellular radiosensitivity. The data support a role for topoisomerase II in the repair of DNA strand breaks. PMID- 11023610 TI - Radiation protection by the ocimum flavonoids orientin and vicenin: mechanisms of action. AB - In previous studies, flavonoids, orientin and vicenin, that were isolated from the leaf extract of Ocimum sanctum, were found to protect mice against radiation injury. Several flavonoids are known to be good antioxidants. Therefore, the effect of orientin and vicenin on radiation-induced lipid peroxidation in vivo and their antioxidant activity in vitro were studied. Adult mice were injected intraperitoneally with 50 microgram/kg of orientin or vicenin and exposed whole body to 3 Gy of gamma radiation. Lipid peroxidation was measured in the liver 15 min to 8 h postirradiation. The antioxidant activity of orientin/vicenin (10-500 microM) was studied by measuring inhibition of hydroxyl radicals generated by the Fenton reaction (Fe(3+)-EDTA-ascorbic acid-H(2)O(2)) in vitro. The compounds were also tested for possible pro-oxidant and iron chelation activities at the above concentrations in the in vitro system. Orientin and vicenin provided almost equal protection against radiation-induced lipid peroxidation in mouse liver. Both compounds showed a significantly greater free radical-inhibiting activity in vitro than DMSO. Neither orientin nor vicenin showed any pro-oxidant activity at the concentrations tested. Both compounds inhibited free radical formation in the absence of EDTA. Free radical scavenging appears to be a likely mechanism of radiation protection by these flavonoids. PMID- 11023612 TI - X- and Q-band electron paramagnetic resonance of CO2- in hydroxyapatite single crystals. AB - Both X- and Q-band electron paramagnetic resonance (EPR) research has been conducted using slightly carbonated hydroxyapatite (HAp) single crystals after exposure to ionizing radiation. Below a temperature of 90 K, O(-) and CO(2-) radicals were detected, whereas at room temperature only CO(2-) spectra could be observed. The O(-) ion has previously been investigated in high-purity HAp single crystals, whereas EPR spectra of CO(2-) in HAp single crystals have not been reported. Both paramagnetic defects exhibit EPR angular variations in planes containing the c axis of the crystal from which spin Hamiltonian parameters were derived. Arguments are given for the presence of two CO(2-) defects in the irradiated HAp single crystals. PMID- 11023613 TI - Human TP53 from the malignant glioma-derived cell lines M059J and M059K has a cancer-associated mutation in exon 8. AB - The protein coding segment of the TP53 genes from the glioma-derived cell lines M059J and M059K was sequenced. The sequences from both cell lines were identical over 5039 bp, including the gene segment containing exons 2 through 9, exon 10, and the proximal segment of exon 11. In both cells, the first nucleotide of codon 286 (GAA, Glu) is changed to an A (AAA, Lys). Comparison with the same TP53 segment from the A549 human lung carcinoma cell line revealed several differences in intron sequence. PMID- 11023614 TI - Glutamate stimulation of raphe pallidus attenuates the cardiopulmonary reflex in anaesthetised rats. AB - Serotonin has been implicated as having a modulatory action on NTS neurones mediating vagal inputs controlling the cardiorespiratory system. Since raphe pallidus and raphe obscurus both send serotonergic projections to the NTS, we have investigated a neuromodulatory role for these structures on the cardiopulmonary reflex. A multibarrelled microelectrode positioned around the level of the area postrema was placed at varying depths into mid-line brainstem structures and the effect of glutamate stimulation on the cardiopulmonary reflex tested. Excitatory chemical stimulation in the region of raphe pallidus, but not raphe obscurus, attenuated significantly the respiratory and bradycardic components of the cardiopulmonary reflex. This attenuation was reversed by an NTS microinjection of RS-39604, a selective 5-HT, receptor antagonist. We propose that neurones in raphe pallidus that project to the NTS can release serotonin which acts via 5-HT4 receptors to attenuate the reflex phrenic nerve activity and heart rate components of the cardiopulmonary reflex. PMID- 11023615 TI - Absence of cholinergic sympathetic innervation from limb muscle vasculature in rats and mice. AB - Although the existence of cholinergic sympathetic vasodilatory innervation in limb muscle vasculature is well established for some species, previous pharmacological studies have failed to reveal the presence of such innervation in rats. Recently, Schafer and colleagues [Schafer, M.K., Eiden, L.E., Weihe, E., 1998. Cholinergic neurons and terminal fields revealed by immunohistochemistry for the vesicular acetylcholine transporter. II. The peripheral nervous system. Neuroscience 84(2), 361-376] reported that vesicular acetylcholine transporter immunoreactivity (VAChT-IR), a marker for cholinergic terminals, is present in the innervation of the microvasculature of rat hindlimb skeletal muscle and concluded that rats possess cholinergic sympathetic innervation of limb muscle vasculature. Because of our interest in identifying targets of cholinergic sympathetic neurons, we have analyzed the transmitter properties of the innervation of muscle vessels in rat and mouse limbs. We found that the innervation of vasculature in muscle is noradrenergic, exhibiting robust catecholamine histofluorescence and immunoreactivity for tyrosine hydroxylase (TH) and the peptide transmitters, neuropeptide Y (NPY) and occasionally vasoactive intestinal peptide (VIP). In contrast, cholinergic phenotypic markers,VAChT-IR and acetylcholinesterase (AChE) activity, are absent. Neuron cell bodies in sympathetic ganglia, retrogradely labeled with injections of tracer into limb muscles, also lacked VAChT but contained TH-IR. The innervation of large extramuscular feed arteries in hindlimbs was also devoid of cholinergic markers, as were the cell bodies of sympathetic neurons innervating extramuscular femoral arteries. These results, like those of previous physiological studies, provide no evidence for the presence of cholinergic sympathetic innervation of muscle vasculature in rats or mice. PMID- 11023616 TI - Potentiation by neostigmine of responses to vagal nerve stimulation in the sinus venosus of the toad. AB - The effects of the cholinesterase inhibitor neostigmine on the responses to vagus nerve stimulation of isolated sinus venosus/atrial preparations of the toad, Bufo marinus, were examined. In control solutions, trains of stimuli applied to the vagus nerve led to a decrease in heart rate that was susceptible to muscarinic receptor blockade. Membrane potential recordings made from sinus venosus cells showed that the responses to trains of stimuli, delivered at frequencies of less than 10 Hz, were little changed by the addition of neostigmine. However, the responses to longer trains of stimuli at 10 Hz (30 versus 10 s) were potentiated and the nature of the membrane potential changes was altered. The results suggest that, due to the activity of cholinesterases, acetylcholine (ACh) released from parasympathetic nerves normally has little access to the muscarinic receptors in the pacemaker region, which are linked to potassium channels. PMID- 11023617 TI - Cardiac vagal responsiveness during development in spontaneously hypertensive rats. AB - In the present study we sought to determine the effect of age, hypertension and endogenous angiotensin on the chronotropic responses to vagal stimulation in urethane anesthetized-normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). After beta-adrenoceptor blockade with atenolol, the right and left vagal nerves were stimulated with graded frequencies between 1 and 32 Hz in 5-, 8- and 22-week-old animals. At all ages and in both strains, there was a strong linear relationship between the degree of bradycardia and the log of the stimulation frequency. At the age of 5 weeks, the bradycardia to stimulation of the right vagus was greater in SHR than that observed in WKY (P<0.05). However, in 8- and 22 week-old animals, no differences were observed between the response to vagal stimulation in WKY and SHR. Thus, there was an age-dependent increase in the response to right vagal stimulation in WKY, but no such trend in SHR. No significant age-dependent changes in left vagal responses were observed in either strain. Left vagal responses were approximately half of the response to right vagal stimulation at all ages in SHR and in 8-22 week WKY, but similar to right vagal responses in 5 week WKY. Administration of the angiotensin converting enzyme inhibitor perindopril, which effectively blocked the formation of endogenous angiotensin, did not affect responses to vagal stimulation at any age, in either strain. These results suggest that the baroreflex vagal deficit observed in adult SHR compared to WKY is not due to a difference in the responsiveness of the cardiac vagal neuroeffector mechanism nor due to an effect of circulating angiotensin II. Furthermore, the enhanced vagal bradycardia observed in very young SHR which was due primarily to the earlier establishment of the adult vagal response pattern may indicate accelerated vagal development in this strain compared to WKY. PMID- 11023618 TI - Mechanism of anemia associated with autonomic dysfunction in rats. AB - The aim of this study was to elucidate the mechanism of anemia associated with autonomic dysfunction in rats. Using 6-hydroxydopamine (6-OHDA)-treated sympathectomized rats, changes in systolic blood pressure, plasma catecholamine levels, hemograms, erythropoietin (EPO) secretion, and beta-adrenergic receptors on erythrocytes were monitored, and compared with desipramine- and 6-OHDA treated, and control rats. In 6-OHDA-treated rats, systolic blood pressure and plasma catecholamine levels significantly decreased from 7 days after 6-OHDA administration, returning to the control values on day 28. Hemoglobin (Hb), hematocrit (Hct) and red blood cell (RBC) levels significantly decreased from day 14 to day 28, and reached normal values after day 35, but neither corpuscular constants nor white blood cell (WBC) levels changed after anemia occurred. Administration of desipramine 1 day before 6-OHDA injection prevented anemia. EPO levels did not elevate, even after bloodletting to load anemia, and the EPO circadian rhythm was irregular in 6-OHDA-treated rats. beta-adrenergic receptors measured using 125I-cyanopindolol (CYP) significantly decreased from day 7 to day 28, and reached normal values after day 35. These results suggest that irregular EPO secretion via disordered autonomic nerves may induce anemia in patients with autonomic disorders. PMID- 11023619 TI - Compensatory vasoconstrictor effects of sodium pentobarbital on the hindquarters of conscious normotensive control and lumbar-sympathectomized Wistar rats. AB - The aim of this study was to compare the vasoconstrictor effect of sodium pentobarbital on the hindquarter resistance of intact control Wistar rats with the effect on lumbar-sympathectomized rats. For this purpose, mean arterial pressure (MAP) and hindquarter (supplied terminal aorta) flow (HQF) were simultaneously measured in these conscious rats with an arterial in dwelling cannula and electromagnetic flow probe implanted around the terminal aorta. Hindquarter resistance (HQR) was calculated as MAP divided by HQF. In the intact control conscious rats, subsequent pentobarbital anesthesia (30 mg/kg, i.v.) caused an increase in HQR (+43.5 +/- 7.4%, mean +/- S.E.M.) and a decrease in MAP (-17.0 +/- 3.2%). After pentobarbital anesthesia, subsequent ganglionic blockade with hexamethonium bromide (C6; 25 mg, i.v.) induced a significant decrease in HQR (-30.9 +/- 3.0%) with a further lowering of MAP (-20.9 +/- 1.6%). However, in rats not anesthetized with sodium pentobarbital, C6 alone induced almost no change in HQR (-3.4 +/- 5.3%), even when MAP was lowered (-24.2 +/- 2.5%). In the lumbar-sympathectomized rats, pentobarbital anesthesia produced almost no change in HQR (-11.7 +/- 4.4%), although MAP decreased significantly (-24.3 +/- 2.2%). These findings suggest that: (1) sodium pentobarbital anesthesia newly generates a compensatory vasoconstrictor tone in the hindquarters acting against the depressor effect, and (2) the vasocompensator tone is controlled by the efferent fibers, including those in the lumbar sympathetic nerves. PMID- 11023620 TI - Changes in regional vascular resistance in response to microinjection of L glutamate into different antero-posterior coordinates of the RVLM in awake rats. AB - Changes in mean arterial pressure (MAP) and in regional vascular resistance (RVR, hindquarter, mesenteric and renal) induced by microinjection of L.-glutamate into three different antero-posterior coordinates of the rostral ventrolateral medulla (RVLM) [1,200-1,600 microm (n=10), 1,601-2,000 microm (n=12) and 2,001-2,500 microm (n=6) rostral to the obex] were investigated in unanesthetized rats. Guide cannulas directed towards the RVLM were implanted 4 days prior to the experiments. Doppler probes were implanted around the superior mesenteric, inferior abdominal aorta and left renal arteries and a catheter was inserted into the femoral artery and vein 1 day prior to the experiments. Insertion of the injector into the RVLM produced an increase in baseline MAP, which was back to control levels 2 min later, when L-glutamate was microinjected. Microinjection of L-glutamate (1 nmol/30 nl) into the three antero-posterior coordinates of the RVLM produced an increase in MAP associated with a similar increase in hindquarter, mesenteric and renal vascular resistance, which were back to control 1 min later. Saline into the RVLM produced negligible effects on MAP and RVR. These findings suggest that the sympathetic vasomotor neurons involved in the regulation of the regional vascular resistance in rats are not topographically distributed in the antero-posterior coordinates of the RVLM. However, the experimental methods used to evaluate the topographic distribution of sympatho vasomotor neurons in the RVLM and the measurement of the regional blood flow may not be precise enough to detect any possible differences. PMID- 11023621 TI - Chronic activation of central alpha2-adrenoceptors prevents hypertension in DOCA salt rats. AB - The effects of chronic intracerebroventricular (i.c.v.) injections of the alpha2 adrenoceptor agonist, xylazine, on blood pressure were examined in DOCA-salt rats. Acute studies also examined the renal sympathetic nerve activity (RSNA) and renal excretory responses produced by i.c.v. xylazine in rats with established DOCA-salt hypertension. Rats implanted with a chronic i.c.v. cannula for drug injection were used. In chronic studies, four groups were investigated: control rats treated with s.c. soybean oil and i.c.v. saline; DOCA-salt rats (s.c. deoxycorticosterone acetate) receiving i.c.v. saline, xylazine or the alpha2 adrenoceptor antagonist, yohimbine. During vehicle or DOCA-salt treatment, xylazine (0.2 ng/microg) or yohimbine (10O microg/kg) was injected i.c.v. daily (three times). In DOCA-salt rats receiving i.c.v. saline, resting mean arterial pressure (MAP) was elevated on days 15 and 30 (135 +/- 5 and 160 +/- 6 mmHg, respectively). Chronic i.c.v. xylazine significantly attenuated the rise in MAP produced by DOCA-salt (day 15, 118 +/- 5 mmHg; day 30, 121 +/- 4 mmHg). Alternatively, chronic i.c.v. yohimbine shortened the onset (day 15, 152 +/- 7 mmHg) and augmented the hypertension in DOCA-salt rats (0 survival by day 30). In acute studies, i.c.v. xylazine elicited a profound natriuresis and diuresis as well as a reduction in RSNA without altering MAP. This study demonstrates that the ongoing (tonic) activity of central alpha2-adrenoceptor mechanisms are critically involved in regulating blood pressure in the DOCA-salt treated rat. In this manner, an enhanced activity of central alpha2-adrenoceptor systems acts to protect against a rise in blood pressure. In contrast, the attenuation of central alpha2-adrenoceptor stimulation evokes hypertension. The central action of xylazine to prevent hypertension may be associated with the inhibition of sympathetic outflow to the kidneys and evokes an enhanced natriuresis. By inhibiting the avid sodium retention elicited by DOCA-salt treatment, the central activation of alpha2-adrenoceptors delays the onset and the severity of hypertension in this pathological model. PMID- 11023622 TI - Recruiting participants for community-based research: the Diabetic Retinopathy Awareness Program. AB - PURPOSE: Recruiting participants is a major challenge for population studies. We present the recruitment methods followed by the Diabetic Retinopathy Awareness Program (DRAP), a community-based, randomized, masked, controlled trial to meet and exceed its sample size goals. METHODS: A county-wide multi-media promotional campaign to recruit and enroll participants in the trial was planned and executed from October 1993 through April 1994, with the assistance of the local news media and community and professional groups. A toll-free 800 number recruitment line was established, and postage-paid recruitment postcards distributed. The trial was designed to examine whether a mailed educational intervention could increase compliance with vision care guidelines among persons with diabetes in the community. RESULTS: A total of 2308 persons with diabetes were interviewed for eligibility and 813 enrolled in the intervention trial, exceeding the original recruitment goals of 1800 and 600, respectively. Those who completed the enrollment interview reflected county demographics. During recruitment, newspaper, television and radio stories featured the study; pharmacies and physician offices displayed study materials; public service announcements appeared in local print and broadcast media. The largest single recruitment response was a local television news report, followed by a newspaper story. CONCLUSIONS: These experiences substantiate the need for a comprehensive coordinated approach, using planned multiple sources, to achieve recruitment success. By engaging the lay and professional communities along with the media, recruitment costs can be kept to a minimum. Participant costs can be minimized by employing a toll-free number and eliminating study participant travel, thus allowing for inclusion of traditionally underserved populations. This approach is applicable to other studies, where community-based participation is desired. PMID- 11023623 TI - Serum ferritin and death from all causes and cardiovascular disease: the NHANES II Mortality Study. National Health and Nutrition Examination Study. AB - PURPOSE: The purpose of this study was to assess the association between serum ferritin and death from all causes, cardiovascular diseases (CVD), CHD and myocardial infarction (MI). Positive body iron stores have been proposed as a risk factor for coronary heart disease (CHD). While most epidemiologic studies using serum ferritin and other measures of body iron stores have not found an association between iron and heart disease risk, the hypothesis remains controversial. As a result, we examined the relationship of serum ferritin, the principle blood measure of body iron stores, to risk of death in a cohort with a standardized exam and long follow-up. METHODS: The baseline data for this prospective cohort study were collected in 1976-1980 as part of the second National Health and Nutrition Examination Study (NHANES II) with mortality follow up using the National Death Index (NDI) through December 31, 1992. The analytic sample (n = 1604) consisted of 128 black men, 658 white men, 100 black women and 718 white women 45-74 years of age at baseline who, based on self-reported data, were free of coronary heart disease at baseline and had no missing data. The main outcome measures were the relative risk of death for persons with serum ferritin levels: <50 microg/L; or 100-199 microg/L; or > or =200 microg/L was compared to persons with serum ferritin levels of 50-99 microg/L adjusted for possible confounding using the Cox proportional hazards model. RESULTS: Most of the deaths were among white men (n = 254) and women (n = 168). There were relatively few deaths among black men (n = 50) and too few in women (n = 23) to reliably model. The largest number of CVD (n = 119), CHD (n = 82), and MI (n = 49) deaths were in white men while there were 69 CVD, 45 CHD and 13 MI deaths in white women. Black men with a serum ferritin level of <50 microg/L had a significantly higher adjusted risk of death from all causes (RR = 3.1 with 95% confidence limits of 1.5-6.5). There were no other statistically significant associations for all causes mortality for the other three race/sex groups. Additionally, there were no statistically significant associations between serum ferritin and any of the cardiovascular endpoints for any of the groups. There was an apparent but nonsignificant u-shaped association between serum ferritin and all causes mortality in black men and between serum ferritin and CVD death in white women. However, in both cases very wide confidence limits preclude further interpretation. CONCLUSIONS: Overall, the results do not support the hypothesis that positive body iron stores, as measured by serum ferritin, are associated with an increased risk of CVD, CHD or MI death or between serum ferritin and all causes mortality. Most of the research to date with serum ferritin has been conducted in European men or in European American men. Our results are consistent with the primarily negative results for that race/sex group. More research is needed in women and minority groups, including an explanation of why such an association would exist in these groups but not in white men before an association can be established in them. PMID- 11023625 TI - Differential regulation of the oscillations in sympathetic nerve activity and renal blood flow following volume expansion. AB - Renal sympathetic nerve activity (RSNA) and renal blood flow (RBF) both show oscillations at various frequencies but the functional significance and regulation of these oscillations is not well understood. To establish whether the strength of these oscillations is under differential control we measured the frequency spectrum of RSNA and RBF following volume expansion in conscious rabbits. Seven days prior to experiment animals underwent surgery to implant an electrode for recording renal nerve activity and a flow probe for recording RBF. Volume expansion (Haemaccel, 1.5 ml min(-1) kg(-1) for 15 min) resulted in a 25 +/- 5% decrease in mean RSNA, paralleled by an increase in RBF to 60 +/- 12 ml min(-1) from resting levels of 51 +/- 11 ml min(-1). Renal denervated rabbits did not show an increase in RBF with volume expansion. Arterial baroreflexes were unaltered by volume expansion. Spectral analysis of the different frequencies in RSNA showed oscillations in RSNA between 0.2 and 0.4 Hz were selectively decreased following volume expansion (14 +/- 3 to 6 +/- 1% of total power in RSNA at < 3 Hz). A corresponding decrease in the strength of oscillations in RBF at this frequency was also seen (20 +/- 6 to 8 +/- 2%). In contrast, the strength of respiratory (0.8-2.0 Hz) and cardiac (3-6 Hz) related rhythms did not change with volume expansion. These results show that selective changes in the different frequency components of RSNA can occur. We suggest that input from cardiopulmonary receptors and/or other vascular beds, and/or altered vascular resistance after volume expansion can reduce the strength of the 0.3 Hz oscillation independent of changes in arterial baroreflex control of RSNA. PMID- 11023624 TI - Age-dependent changes of electrophysiologic characteristics of the stellate ganglion conducting pathways in kittens. AB - The conducting pathways of the cat stellate ganglia were examined in newborn, 10- and 20-day-old kittens by recording the evoked responses in the branches of the ganglion following electrical stimulation of its other branches. In the course of postnatal ontogenesis, the average conduction velocity of excitation and the average amplitude of the responses was increased. In newborn and 10-day-old kittens, all fibers on their conduction velocities belong to C-fibers. In 20-day old kittens, Adelta- and B-fibers also appeared. In this age-group, two subgroups among fibers of the C group were determined according to their conduction velocity of excitation. From 10 days of age, the axon reflex was recorded in the anastomosis and the inferior cardiac nerve following stimulation of the cranial and caudal branches of the subclavian loop. All branches of the stellate ganglia of 20-day-old kittens contain both continuous and synaptically switched fibers. PMID- 11023626 TI - Abdominal vagi mediate c-Fos expression induced by X-ray irradiation in the nucleus tractus solitarii of the rat. AB - The mechanism of induction of emesis by X-ray irradiation remains largely unknown. The purpose of the present research was to clarify the neuronal basis of the induction of nausea induced by X-ray irradiation analyzing c-Fos expression in the nucleus tractus solitarii (NTS) as a marker of cellular excitation. We confirmed that the dose of X-ray irradiation (4 Gy) used for the present research could actually induce nausea by preliminary measurement of kaolin intake. Induction of c-Fos immunoreactivity in the NTS was observed in the animals that received X-ray irradiation of the whole body. The mean number of c-Fos positive cells in the animals that received irradiation was significantly larger than that in the non-irradiated animals. Partial exposure of the abdomen to X-rays showed significantly greater c-Fos expression than that of the head. These results indicated the presence of a certain route for transmitting information from the periphery toward the central nervous system by X-ray irradiation. The number of c Fos positive cells induced by X-ray irradiation in animals vagotomized at the subdiaphragmatic level was lower than that in sham-operated animals. Animals receiving a serotonin subtype three (5-HT3, 5-hydroxytryptamine) receptor antagonist (tropisetron, ICS 205-930, 3-tropanyl-indole-3-carboxylate) showed a significant reduction in c-Fos protein expression compared to animals receiving a vehicle. These results strongly suggested that X-ray irradiation activates 5-HT3 receptors on the terminals of the abdominal vagal nerves to excite the afferent pathway, thereby inducing emesis. PMID- 11023627 TI - Roles of central glutamate, acetylcholine and CGRP receptors in gastrointestinal afferent inputs to vagal preganglionic neurones. AB - It is unknown which neurotransmitter receptors are involved in the transfer of sensory information from the digestive tract to the brainstem. We examined the potential involvement of central glutamate, acetylcholine, and CGRP receptors in vagal pathways arising from gut chemo- and mechanosensitive afferents. Unitary recordings were made from 21 vagal preganglionic efferent neurones. Neuronal discharge showed either excitation or inhibition in response to oesophageal balloon distension (1-2 ml), gastric distension (40-60 ml in whole stomach or 20 ml in proximal stomach), cholecystokinin-8s (100 pmol close intra-arterially) and bradykinin (18 nmol close intra-arterially). Effects of glutamatergic non-NMDA, muscarinic M1, and CGRP1 receptor antagonism on efferent responses were investigated using CNQX (75-155 nmol i.c.v.), pirenzepine (2.5-5.0 micromol/kg i.v.), and hCGRP8-37 (3.2-6.4 nmol i.c.v.), respectively. CNQX, pirenzepine, and hCGRP8-37, respectively, altered efferent responses in 65%, 23% and 41% of neurones. When both CNQX and hCGRP8-37 were administered, a further 58% of responses were reduced. CNQX and hCGRP8-37 reduced a proportion of efferent responses to all stimuli whereas pirenzepine selectively reduced only efferent responses to gastric distension. We conclude that central CGRP1 and non-NMDA receptors are involved in mediating a range of upper gastrointestinal mechano- and chemo-sensitive afferent inputs onto vagal efferents. M1 receptors, on the other hand, are selectively involved in neurotransmission from gastric mechanoreceptors. PMID- 11023628 TI - Autonomic regulation of cystatin S gene expression in rat submandibular glands. AB - Innervation of rat submandibular and parotid glands by the autonomic nervous system regulates saliva volume, its rate of secretion and its composition. The autonomic nervous system also plays a regulatory role in the differentiation and growth of salivary glands, and in the expression of specific sets of genes. Rat cystatin S, a member of family 2 of the cysteine proteinase inhibitor superfamily, is expressed in submandibular and parotid glands of human and rat. In the rat, cystatin S gene expression is tissue- and cell type-specific, is temporally regulated during postnatal development, and not observed in adult animals. The beta-adrenergic agonist isoproterenol (IPR) induces hypertrophic and hyperplastic enlargements of rat salivary glands and the expression of a number of genes including cystatin S. Sympathectomy reduces, but does not completely block, IPR-induced expression of the cystatin S gene in submandibular glands of adult female rats, indicating the participation of sympathetic factor(s) in its regulation. Bilateral parasympathectomy also reduces IPR-induced cystatin S gene expression, suggesting a role of the parasympathetic nervous system in its regulation. Experiments described in this paper suggest that similar factor(s) arising from both the sympathetic and parasympathetic branches of the autonomic nervous system simultaneously participate in IPR-induced cystatin S gene expression in submandibular glands. PMID- 11023629 TI - Vasoactive intestinal polypeptide and calcitonin gene-related peptide in the developing rat heart atria. AB - Vasoactive intestinal polypeptide-like immunoreactivity (VIP-LI) and calcitonin gene-related peptide (CGRP)-LI concentrations were determined in the developing rat heart atria using radioimmunoassay. Peptide levels were analysed on postnatal days 1, 10, 25, 45, 60, and 85 (P1-P85) separately in the right (RA) and left atria (LA). No sex differences were revealed at any age examined. VIP-LI has been already detected in both atria at P1 in concentrations comparable to values at P10. In the RA, VIP-LI levels increased significantly between days P10 and P25, remained high at P45 and then declined. In the LA, VIP-LI concentrations did not differ from those in the RA on days P1, P10, P25, and P45. However, regional differences were found at P60 and P85, when the peptide levels were significantly higher in the LA than in the RA. The postnatal changes in CGRP-LI concentrations were comparable in both atria with similar values at P1 and P85. After birth, CGRP levels decreased gradually till P45, then they increased till P60 and declined again at P85. The results demonstrate that there is an asymmetry in the postnatal development of the atrial VIP-LI and CGRP-LI concentrations. VIP-LI levels reached their maximum at P25, whereas CGRP-LI levels at P60. Relatively high peptide concentrations in neonatal atria and their variations during development might be related to diverse trophic functions of VIP and CGRP. PMID- 11023630 TI - Reflex patterns in preganglionic sympathetic neurons projecting to the superior cervical ganglion in the rat. AB - Reflex patterns in preganglionic neurons projecting in the cervical sympathetic trunk (CST) were analyzed in response to stimulation of various afferent systems. We focused on the question whether these preganglionic neurons can be classified into functionally distinct subpopulations. Reflex responses were elicited by stimulation of trigeminal and spinal nociceptive, thermoreceptive as well as baroreceptor and chemoreceptor afferents. Multi- and single fiber preparations were studied in baroreceptor intact and sino-aortically denervated animals. Spontaneous activity of 36 preganglionic single neurons ranged from 0.2 to 3.5 imp/s (median= 1.11 imp/s). The degree of cardiac rhythmicity (CR) in the activity of sympathetic neurons was 69.5+/-13% (mean+/-S.D.; N=52; range=39-95%). Noxious stimulation of acral skin activated the majority (67%) of sympathetic preparations by 37+/-25% (N=35) above pre-stimulus activity; 15% were inhibited. In these neurons the response to noxious stimulation of acral skin was significantly correlated with the degree of CR (P<0.001, N=52) in that neurons showing the strongest excitation to noxious stimulation displayed the strongest CR. Noxious mechanical stimulation of body trunk skin (N=60) inhibited the majority (80%) of fiber preparations tested (by 34+/-18% of pre-stimulus activity, N=48); an activation was not observed. Cold stimulation of acral (N=9) and body trunk skin (N=42) activated most fiber preparations. Trigeminal stimulation evoked a uniform reflex activation of preganglionic neurons (+79+/ 73% of pre-stimulus activity, N=32). Chemoreceptor stimulation by systemic hypercapnia elicited inhibitory (-31+/-19%, N=8) as well as excitatory (+59+/-5%, N=4) responses. These results show that preganglionic sympathetic neurons projecting to target organs in the head exhibit distinct reflex patterns to stimulation of various afferent systems; however, a clear classification into different functional subgroups did not emerge. Furthermore, reflex patterns showed a segmental organization to noxious cutaneous stimulation of acral parts and body trunk reflecting a differential central integration of spinal afferent input. Compared with the cat the reflex organization of sympathetic neurons projecting to the head seems to be less differentiated in the anesthetized rat. PMID- 11023631 TI - Respiratory rhythmicity in the activity of postganglionic neurones supplying the rat tail during hyperthermia. AB - It has been suggested that thermoregulatory stimulation changes respiration related rhythmicity in the activity of postganglionic sympathetic neurones supplying the rat tail to a distinct modulation independent of respiration. To study this possibility, single and few fibre recordings were made from ten filaments split from the ventral collector nerves of the rat during whole body warming. Sympathetic activity was analysed by autocorrelation and phrenic triggered summation. All neurones except one were gradually inhibited and lost their on-going activity above a core temperature of 39-39.5 degrees C while the frequency of the phrenic bursts increased significantly. During hyperthermia, all neurones tested exhibited a prominent respiratory modulation in their activity which, compared to normothermia, was significantly increased in strength, or even newly acquired. No other rhythm emerged. These results speak against the hypothesis that in the rat sympathetic pathways controlling the tail vasculature and thus involved in thermoregulation, during hyperthermia become controlled by central oscillators distinct from the respiratory rhythm generator. Rather, respiratory modulation appears to remain the dominant rhythm as is common for sympathetic neurones supplying other cardiovascular targets. PMID- 11023632 TI - Identification of intestinofugal neurons projecting to the coeliac and superior mesenteric ganglia in the rat. AB - Intestinofugal neurons are parts of the afferent limbs of inhibitory intestino intestinal reflexes. These neurons have been mapped in guinea-pigs, where they have a gradient of increasing frequency of occurrence from oral to anal, but not in other species. In the present work in the rat, a species that is more amenable to physiological study than the guinea-pig, we have used retrograde tracing to map the distribution of the cell bodies of intestinofugal neurons projecting to the coeliac-superior mesenteric ganglion complex. Labelled nerve cells were found in the myenteric, but not the submucosal plexus. They were mono-axonal neurons, most with Dogiel type I morphology, and were immunoreactive for choline acetyltransferase, implying that they are cholinergic, which is consistent with functional studies. The cells increased in number per unit area from the stomach, through the small intestine, to the caecum. The results are consistent with physiological studies that reveal distal to proximal inhibitory reflexes that are more potent from distal compared to proximal sites. PMID- 11023633 TI - Micturitional disturbance in neuro-Behcet's syndrome. AB - BACKGROUND: Micturitional disturbance is known to occur in neuro-Behcet's syndrome (NBS). However, its pathophysiology is uncertain and few data are available concerning urodynamic findings. We describe our findings on micturitional histories and urodynamic studies in patients with NBS. METHODS: A history of urinary symptoms was obtained from nine patients with NBS [seven male, two female, aged 26 to 54 years; mean, 41 years]. Neurological and CT/MRI findings indicated that their lesions were located mainly in the cerebral cortex, basal ganglia, cerebellum, brainstem and the spinal cord. Symptomatic patients underwent urodynamic studies, including measurement of post-micturition residuals, urethral pressure profilometry and EMG cystometry. RESULTS: Six of the nine patients (67%) had urinary symptoms that included diurnal urinary frequency in five, sensation of urgency in five, urge urinary incontinence in five, voiding difficulty in five and nocturnal urinary frequency in four. None had urinary retention. Urodynamic studies performed in six patients showed post-micturition residuals in five (50-180 ml), increased maximum urethral closure pressure in two, decrease in four and increase in one of bladder capacities, detrusor hyperreflexia in five, decreased bladder sensation in one, brisk bulbocavernosus reflex in four, detrusor-sphincter dyssynergia in two and neurogenic sphincter EMG in one. CONCLUSION: Our results indicate that micturitional disturbance is not rare in patients with NBS. Supranuclear pelvic nerve dysfunction seems to be mainly responsible for the disturbance. PMID- 11023634 TI - Three-dimensional return map: a new tool for quantification of heart rate variability. AB - BACKGROUND: Several methods are used to study heart rate variability, but they have limitations, which might be overcome by the use of a three-dimensional return map. OBJECTIVES: To evaluate the performance of three-dimensional return map-derived indices to detect (1) sympathetic and parasympathetic modulation to the sinus node and (2) autonomic dysfunction in diabetic patients. METHODS: Six healthy subjects underwent partial and total pharmacological autonomic blockade in a protocol that incorporated vagal and sympathetic predominance. Twenty-two patients with type 2 diabetes mellitus and 12 normal controls participated in the subsequent validation experiment. Three-dimensional return maps were constructed by plotting RRn intervals versus the difference between adjacent RR intervals [(RRn+1)-(RRn)] versus the number of counts, and four derived indices (P1, P2, P3, MN) were created for quantification. RESULTS: Both indices P1 and MN were significantly increased after sympathetic blockade with propranolol, while all indices except P1 were modified after parasympathetic blockade (P < 0.05). During the validation experiments, P1 and MN detected differences between normal controls, and diabetic patients with and without autonomic neuropathy. The overall accuracy of most three-dimensional indices to detect autonomic dysfunction, estimated by the area under the ROC curve, was significantly better than traditional time domain indices. Three-dimensional return map-derived indices also showed adequate reproducibility on two different recording days (intra-class correlation coefficients of 0.69 to 0.82; P < 0.001). CONCLUSIONS: Three-dimensional return map-derived indices are reproducible, quantify parasympathetic as well as sympathetic modulation to the sinus node, and are capable of detecting autonomic dysfunction in diabetic patients. PMID- 11023635 TI - Visuographemic alexia: a new form of a peripheral acquired dyslexia. AB - We report a single-case study of peripherally acquired dyslexia that meets the clinical criteria of "alexia without agraphia." The patient, AA, has a large infarct involving the left posterior cerebral artery. The most striking feature is a severe impairment in recognizing single visually presented letters that precludes explicit or implicit access to reading, even in a letter-by-letter fashion. AA can, however, differentiate letters from similar nonsense characters and digits, and he is also able to identify alphanumeric signs when the visual channel is bypassed (through somesthesic or kinesthesic presentation). Spelling tasks are also well performed. Since there is a breakdown in mapping a visually presented letter to its abstract graphemic representation, we propose the term "visuographemic alexia" for this kind of reading disorder. The pattern of deficits is interpreted following theoretical models previously developed in cognitive neuropsychology. An alexia for arabic numerals with preserved comprehension lends additional support for the crucial processing of different notational systems (e.g., phonographic vs logographic). More general perceptive disorders do not seem to account for these patterns; they are material-specific. Finally, we attempt to specify functional correlations with the implied neural networks. PMID- 11023636 TI - Oral dyspraxia in inherited speech and language impairment and acquired dysphasia. AB - Half of the members of the KE family suffer from an inherited verbal dyspraxia. The affected members of the family have a lasting impairment in phonology and syntax. They were given various tests of oral praxis to investigate whether their deficit extends to nonverbal movements. Performance was compared to adult patients with acquired nonfluent dysphasia, those with comparable right hemisphere lesions, and age-matched controls. Affected family members and patients with nonfluent dysphasia were impaired overall at performing oral movements, particularly combinations of movements. It is concluded that affected members of the KE family resemble patients with acquired dysphasia in having difficulties with oral praxis and that speech and language problems of affected family members arise from a lower level disorder. PMID- 11023637 TI - Pitch and timing abilities in inherited speech and language impairment. AB - Members of the KE family who suffer from an inherited developmental speech-and language disorder and normal, age-matched, controls were tested on musical abilities, including perception and production of pitch and rhythm. Affected family members were not deficient in either the perception or production of pitch, whether this involved either single notes or familiar melodies. However, they were deficient in both the perception and production of rhythm in both vocal and manual modalities. It is concluded that intonation abilities are not impaired in the affected family members, whereas their timing abilities are impaired. Neither their linguistic nor oral praxic deficits can be at the root of their impairment in timing; rather, the reverse may be true. PMID- 11023638 TI - Pitch and timing abilities in adult left-hemisphere-dysphasic and right hemisphere-damaged subjects. AB - The production and perception of pitch and rhythm were tested in patients with acquired unilateral left-hemisphere (LH) lesions (and subsequent motor dysphasia, n = 13), patients with unilateral right-hemisphere (RH) lesions (n = 14), and normal age-matched controls. While the LH dysphasic subjects were not generally impaired on the production or perception of pitch, they were grossly impaired on the production and perception of rhythm. The RH subjects, in contrast, were impaired on measures of pitch perception and production, including the discrimination and production of single notes and of melodies. It is concluded that the two hemispheres differ in their specialization for the perception and production of pitch and rhythm. PMID- 11023639 TI - Rate of information segregation in developmentally dyslexic children. AB - Slowed processing of sequential perceptual information is related to developmental dyslexia. We investigated this unimodally and crossmodally in developmentally dyslexic children and controls ages 8-12 years. The participants judged whether two spatially separate trains of brief stimuli, presented at various stimulus onset asynchronies (SOA) in one or two senses, were synchronous or not. The stimulus trains consisted of light flashes in vision, clicks in audition, and indentations of the skin in the tactile sense. The dyslexic readers required longer SOAs than controls for successful performance in all six comparisons. The crossmodal spatiotemporal resolution of the groups differed more than unimodal performance. The dyslexic readers' segregation performance was also less differentiated than that of the controls. Our results show that not only sensory but also polysensory nonverbal information processing is temporally impaired in dyslexic children. PMID- 11023640 TI - Exploring the cognitive basis of right-hemisphere pragmatic language disorders. AB - Despite considerable interest in the linguistic dimensions of right-hemisphere (RH) pragmatic language disorders, the cognitive bases for these are rarely examined. This study investigated two alternative explanations. First, RH pragmatic language disorders may reflect failure of the RH to synthesise incoming and preexisting information, verbal and visuospatial. In this case language and visuospatial performance should covary. Alternatively such disorders may reflect damage to executive control of all cognitive processing secondary to frontal system failure. In this case language and executive function would be associated. Further, in the former case, subjects should be insensitive to the plausibility of information, whereas in the latter they would be fixated by the literal meaning of information and therefore highly sensitive to plausibility. Eighteen patients with RH damage were compared to 20 matched controls on a range of language and neuropsychological tasks. Pragmatic performance was generally correlated to RH (visuospatial) function, not to executive function. Nonetheless RH patients were found to have problems ignoring plausibility. Thus the specific RH hypothesis described needs to be reconsidered. PMID- 11023641 TI - Hemispheric specialization for reading. AB - Behavioral laterality tasks with linguistic stimuli were used to assess the differential processing efficiencies of the cerebral hemispheres in right- and left-handed adults. Findings from a lateralized lexical decision task with concrete nouns supported Zaidel's (1983) "direct access" model of hemispheric functioning. A dual task consisting of oral and silent reading indicated that the right hand was significantly more disrupted than the left during unimanual finger tapping; however, some bilateral interference was observed. Taken together the findings suggest that although the left hemisphere was relatively more efficient, the right hemisphere was dynamically involved in the reading process. PMID- 11023642 TI - Category-specific naming errors in normal subjects: the influence of evolution and experience. AB - The importance of "artifactual" variables (such as conceptual familiarity) have been highlighted in current accounts of category-specific disorders for living things (e.g., Funnell & Sheridan, 1992). The difficulties experienced by patients are essentially viewed as an exaggeration of normal processes and the implication is that normal subjects should also have greater difficulty naming living items (because they have lower conceptual familiarity than nonliving things). The current study examined normal subjects' ability to name pictures of artifact matched sets of living and nonliving things in a naming-to-deadline paradigm. Contrary to the prediction, normal subjects made more nonliving naming errors. Furthermore, female subjects made more nonliving-thing errors than male subjects. These findings could not be reduced to differences in either category-based or gender-based familiarity ratings. Rather, it is proposed that an elaborated domain-specific evolutionary model parsimoniously explains both the greater incidence of living thing deficits in patients and the better performance of normal subjects with living things. PMID- 11023643 TI - Cytokines in diseases of the endocrine system. AB - Cytokines, the polypeptide mediators of the immune system, were shown to exert numerous actions on endocrine functions. Bidirectional links based on the sharing of mediators and receptors between the immune and neuroendocrine systems lead to the concept of the immune-neuroendocrine system that seems to constitute an important and sophisticated regulatory system in the homeostasis. Several cytokines were found to be involved in the pathogenesis of diseases of the endocrine system. In this brief review, we attempt to present a general outline of the local actions of cytokines on cells of endocrine organs with an emphasis on disease etiology (pituitary tumours and autoimmune endocrine diseases in particular). PMID- 11023644 TI - Comparison between the in vivo and in vitro expression of three adhesion signaling proteins in embryonic cells. AB - We have examined the relationship between the in vivo and in vitro expression of three adhesion-signaling proteins (FAK, PYK2 and Paxillin), using cells of the early chick embryo, where pure cell populations may be isolated and cultured, and in which epithelial-to-mesenchymal transformation is occurring. Focal Adhesion Kinase (FAK) and Proline-rich Tyrosine Kinase-2 (PYK2) are related in molecular structure, and may have some overlapping functions in signal transduction associated with cell-substratum adhesion. Paxillin, a cytoskeletal protein, is also localized to focal adhesions. We show that the immunocytochemical detection of these molecules in vivo does not reflect their in vitro localization. Focal Adhesion Kinase showed a developmentally regulated localization to the cytoplasm, but not to sites of adhesion, in epithelial cells in vivo, while Paxillin was associated with migrating mesoderm cells. Proline-rich Tyrosine Kinase-2 was undetectable in vivo. The level of expression of these molecules was compared under in vivo and in vitro conditions. While the expression of Focal Adhesion Kinase showed a tissue-specific regulation of expression with the change to in vitro conditions, Proline-rich Tyrosine Kinase-2 showed a more uniform and less tissue-specific up-regulation. Levels of Paxillin expression also showed an increase with this change in conditions. We conclude that despite the structural and functional relationships between these three molecules in the developing embryo, the expression and localization of each is independently regulated. We suggest that this provides these cells with the adaptability that they require in order to respond to the changing extracellular environment in the early embryo, and to undergo epithelial-to-mesenchymal transformation. PMID- 11023645 TI - Differential responsiveness of prostatic acid phosphatase and prostate-specific antigen mRNA to androgen in prostate cancer cells. AB - Androgens regulate the expression of both human prostatic acid phosphatase (PAcP) and prostate-specific antigen (PSA), two major prostate epithelium-specific differentiation antigens. Due to the important role of these two enzymes as prostate epithelium differentiation markers, we investigated their regulation of expression at the mRNA level in LNCaP human prostate carcinoma cells. Interestingly, phenol red, a pH indicator in the culture medium, promoted cell growth. To eliminate this non-specific effect, a phenol red-free, steroid-reduced medium was utilized. When high-density cells were grown in that medium, 5alpha dihydrotestosterone (DHT) suppressed PAcP but stimulated PSA. However, tumor promoter phorbol ester 12-o-tetradecanoyl phorbol-13-acetate (TPA) functioned as a potent inhibitor of both PAcP and PSA expression. Prolonged treatment with DHT as well as TPA resulted in a similar down-regulation of protein kinase C and cellular PAcP activities. Thus, the levels of PAcP and PSA mRNA are differentially regulated by androgens in LNCaP cells. PMID- 11023646 TI - C/EBPalpha and C/EBPbeta are persistently associated with the rat liver nuclear matrix throughout development and the acute phase response. AB - The partitioning of C/EBPalpha and C/EBPbeta on the nuclear matrix structure was examined during the different transcriptional activities accompanying liver development and the acute phase (AP) response. The presence of C/EBPalpha and C/EBPbeta was established on the nuclear matrix. Their relative concentrations on the matrix always reflected the developmental stage- and AP-related fluctuations observed in the nuclear extract. Thus, they progressively increased as development proceeded, whereas during the AP response, C/EBPalpha decreased and C/EBPbeta increased. In addition, the levels of both transcription factors were always notably higher in the nuclear matrix than in the extracts. We conclude that the observed changes and overall enrichment of the nuclear matrix with regulatory proteins is a reflection of the importance of such interactions for the in vivo functioning of C/EBP proteins. PMID- 11023647 TI - Influence of membrane fluidity modifiers on lysosomal osmotic sensitivity. AB - Since lysosomes are prone to osmotic lysis, we have examined the correlation between their physical state and sensitivity to osmotic challenge, using agents which modify membrane fluidity. The latency loss of beta-hexosaminidase after an incubation in hypotonic sucrose medium was followed under different conditions of membrane fluidity, recorded by steady-state fluorescence anisotropy of 1,6 diphenyl-1,3, 5-hexatriene. Increasing fluidity of the lysosomal membranes with benzyl alcohol (BA) and greater rigidity caused by cholesteryl hemisuccinate (CHS) increased and decreased the enzyme latency loss, respectively. The effects of BA and CHS treatments on osmotic sensitivity were reversible subsequently by reciprocal treatments of the lysosomes with CHS and BA, respectively. The results indicate that the physical state of the membrane does indeed affect lysosomal osmotic stability. PMID- 11023648 TI - Catalase activity in human erythrocytes: effect of phenoxyherbicides and their metabolites. AB - The effects of exposure to different concentrations of phenoxyherbicides and their metabolites were studied in human erythrocytes, with particular attention to catalase (CAT-EC. 1.11.1. 6- hydrogen peroxide: hydrogen peroxide oxidoreductase). 4-chloro-2-methylphenoxyacetic acid (MCPA), 2,4-dimethylphenol (2, 4-DMP) and 2,4-dichlorophenoxyacetic acid (2,4-D) did not affect CAT activity, but 2,4-dichlorophenol (2,4-DCP) and 2,4,5-trichlorophenol (2,4,5-TCP) decrease its activity, the latter being the more inhibitory. PMID- 11023649 TI - Different physiological mechanisms control isovolumetric regulation and regulatory volume decrease in chick embryo cardiomyocytes. AB - Cultured chick embryo cardiac myocytes submitted to a 180 mOsm/kg hyposmotic solution swell present a regulatory volume decrease (RVD). This RVD is mediated by a Ca(2+)influx followed by a 40% loss of total taurine content accompanied by the loss of lesser amounts of other osmolytes. Kidney cells respond to a gradual change in osmolality by maintaining their volume at the initial level. This is termed isovolumetric regulation (IVR), which may activate regulatory processes other than those observed with sudden changes in osmolality. When cardiac myocytes were exposed to a gradual change in osmolality, they show a partial IVR which is not dependent upon extracellular Ca(2+). Potassium channel blockers, quinidine and Ba(2+), and the chloride channel blocker, diphenylamine-2 carboxylate (DPC), compromise IVR in our model. Tritiated taurine loss and total intracellular K(+)contents were analyzed in cultured cardiomyocytes submitted to a gradual change in osmolality. The cultured cells lost approximately 10% of their taurine and 35% of their total K(+). These findings suggest that different compensatory mechanisms are activated when cells are exposed to stepwise and gradual changes in osmolality. Inorganic osmolytes (through conductive pathways) are preferentially mobilized during the physiological and/or patho-physiological IVR situation, perhaps reflecting energetic conservation in response to a less traumatic event for the cardiac myocytes. PMID- 11023650 TI - Extended chromatin fibres: crystallinity, molecular order and reactivity to concanavalin-A. AB - The present study was undertaken to test the reproducible formation of the extended chromatin fibres (ECF), beads and superbeads, and detect molecular order and crystallinity by optical anisotropies in those structures. Chicken erythrocyte smears and mouse liver cell imprints were treated with 2.0-3.0 m NaCl solution in 1% Triton-X100 vertically prior to staining with 0.025% toluidine blue at pH 4. Detection of birefringence and colours of abnormal dispersion of birefringence (ADB) following toluidine blue binding to DNA revealed that the DNA molecular order and crystallinity in decondensed and condensed chromatin are preserved after ECF formation. The tests for Con-A binding to mannose/glucose residues of glycoproteins was confirmed within nuclei, and in the ECF, beads and superbeads. ECF formation was not regular. Clumped cells did not show ECF, although chromatin mobility was observed within the nuclei. Electron microscopy demonstrated that after treatment of nuclei with 0.77 m NaCl ECF always spread from the nuclei, in clumped nuclei the fibres aggregated instead of spreading. PMID- 11023651 TI - Radial spindle and the phenotype of the maize meiotic mutant, dv. AB - The morphological phenotype of the maize meiotic mutant dv (divergent spindle) has been further analysed by visualization of the division spindle and examination of its fine structure in mother cells of pollen. Previous research showed that dv blocks convergence of spindle fibres at the poles. New observations reveal abnormalities caused by this mutation, with dv showing disturbances in nuclear envelope breakdown during vesiculation, preventing the spindle fibres from adopting a bipolar orientation (with convergence on the poles). The anomalies result in radial spindles which are similar to monoastral spindles in animal cells. PMID- 11023652 TI - Modes of cell death in the hypopharyngeal gland of the honey bee (Apis mellifera l). AB - Different modes of cell death have been revealed in the regressing hypopharyngeal glands of worker honey bees. The hypopharyngeal gland, which is well developed in young nursing bees to produce protein for larval food, was seen to regress naturally in foraging adult worker bees. A range of techniques including histology, cytochemistry, in situ TUNEL, Annexin V and Comet assays indicated that cells within the gland demonstrate progressive symptoms of apoptosis, necrosis and a vacuolar form of programmed cell death. The latter mode of cell death did not display chromatin margination, but was accompanied by an enhanced level of autophagic and hydrolytic activity in which a cytosolic source of acid phosphatase became manifest in the extra-cisternal spaces. Normal and annexin positive cells were found to occur in the younger nursing bees, whilst necrosis and an aberrant vacuolar type of apoptosis predominated in the older foraging bees. The relevance of these results to the classification of programmed cell death is discussed. PMID- 11023653 TI - Translocation of P53-regulated laminin receptors in pro-apoptotic microcircumstance of human vasculogenesis inhibition. AB - It is not clear what type of cells will inhibit vasculogenesis and under what circumstances. Here we examined the human spontaneous abortion villi and discovered that cells in the vasculogenesis-inhibited environment appear to be pro-apoptotic as indicated by strong staining of bak. In particular, an increasing laminin receptor (LNR) expression in cytoplasm and on apical membrane, being most likely mediated by p53 because of coexpression, was observed in the pro-apoptotic syncytiotrophoblasts. The transcriptional increase and translocation of the receptors to the cytoplasm and apical membrane may be used as a pathological marker, indicating a change of the structure and function of integrins in the non-vasculogenesis micro-environment. PMID- 11023654 TI - Presence, uptake and localization of an immunoreactively interleukin 6 (IL-6) like molecule in Tetrahymena pyriformis. AB - The unicellular Tetrahymena and its medium contain immunoreactively interleukin 6 (IL-6)-like molecules (hereinafter IL-6) in a measurable quantity in the 24 h-old cultures. This protozoan takes up exogenously supplied IL-6 very quickly, and this can be found in similar amounts in both the cells and the media after 1 h. After 24 h (48 h cultures), an equal amount of IL-6 is present in the control and IL-6-treated cells and their media. By 120 h, cells which have not had their medium changed retained the same quantity of IL-6 as the control; however less than half was found in IL-6-treated cells. In the medium of 120 h-old cultures, there was a reduction of IL-6 content relative to the 24 h content in the control; however, in the IL-6-treated cell culture medium, less than half of the level in the controls was found. Confocal microscopy demonstrated the localization of IL-6 in/on the oral apparatus and basal bodies, and the nuclear envelope also showed moderate labelling. IL-6 antibody binding was enhanced after IL-6 pretreatment (hormonal imprinting). The experiments call attention to the presence of an IL-6-like molecule and its uptake at a very low level of phylogeny. PMID- 11023655 TI - 2,7-dichlorofluorescin oxidation and reactive oxygen species: what does it measure? AB - Intracellular 2,7-dichlorofluorescin (H(2)DCF) oxidation is often used to measure the production of reactive oxygen species (ROS) within cells. The rate of H(2)DCF oxidation depends on the concentration of glutathione, which is an alternative target for ROS. Our results suggest that increased rate of H(2)DCF oxidation be interpreted as an indication of general oxidative stress due to a variety of reasons, including depletion of antioxidants, rather than as a specific proof of augmented ROS formation. PMID- 11023656 TI - Draw It again Sam: the effect of drawing on children's suggestibility and source monitoring ability. AB - Preschool children (aged 3 to 6 years) participated in a magic show. Later, the children were given repeated true and false reminders about the show. Half the children were asked to draw these true and false reminders (drawing condition) and half the children were asked questions about the reminders but not to draw them (question condition). Later, children in the drawing condition had better recall of true reminders than children in the question group; however, children in the drawing group also recalled more false reminders than children in the question group. Finally, although children in the drawing group had better memory of the source of the reminders than children in the question group, both groups equally reported that the false reminders actually happened. PMID- 11023657 TI - Individual differences in gains from computer-assisted remedial reading. AB - Two hundred second- to fifth-grade students (aged approximately 7 to 11 years) spent 29 h in a computer-assisted remedial reading program that compared benefits from accurate, speech-supported reading in context, with and without explicit phonological training. Children in the "accurate-reading-in-context" condition spent 22 individualized computer hours reading stories and 7 small-group hours learning comprehension strategies. Children in the "phonological-analysis" condition learned phonological strategies in 7 small-group hours, and divided their computer time between phonological exercises and story reading. Phonologically trained children gained more in phonological skills and untimed word reading; children with more contextual reading gained more in time-limited word reading. Lower level readers gained more, and benefited more from phonological training, than higher level readers. In follow-up testing, most children maintained or improved their levels, but not their rates, of training gains. Phonologically trained children scored higher on phonological decoding, but children in both conditions scored equivalently on word reading. PMID- 11023658 TI - Numerical and arithmetical cognition: a longitudinal study of process and concept deficits in children with learning disability. AB - Based on the stability and level of performance on standard achievement tests in first and second grade (mean age in first grade = 82 months), children with IQ scores in the low-average to high-average range were classified as learning disabled (LD) in mathematics (MD), reading (RD), or both (MD/RD). These children (n = 42), a group of children who showed variable achievement test performance across grades (n = 16), and a control group of academically normal peers (n = 35) were administered a series of experimental and psychometric tasks. The tasks assessed number comprehension and production skills, counting knowledge, arithmetic skills, working memory, the ease of activation of phonetic representations of words and numbers, and spatial abilities. The children with variable achievement test performance did not differ from the academically normal children in any cognitive domain, whereas the children in the LD groups showed specific patterns of cognitive deficit, above and beyond the influence of IQ. Discussion focuses on the similarities and differences across the groups of LD children. PMID- 11023659 TI - The developing role of cytokines for imaging inflammation and infection. AB - The diagnosis of inflammatory processes is an important goal in medicine. In some cases the diagnosis is easy, based on the clinical history and the physical examination of the patient. Other cases are more difficult to diagnose because they are asymptomatic or with non-specific symptoms. Thus, several imaging techniques have been developed for the diagnosis of inflammatory processes, from the simple X-ray to the more sophisticated computerised tomography, magnetic resonance imaging and nuclear medicine scan. They provide different information and their role in different diseases will be discussed in this review with particular emphasis on the expanding field of the use of radiolabelled cytokines for imaging infection/inflammation. So far, IL-1, IL-1ra, IL-2, IL-6, IL-8, IL 10, IL-12 p40, G-CSF, IFN-gamma and EGF have been radiolabelled for in vivo targetting of different leukocyte subsets with promising results for their clinical use. PMID- 11023660 TI - gp130 Cytokine family and bone cells. AB - Bone tissue is continually being remodelled according to physiological circumstances. Two main cell populations (osteoblasts and osteoclasts) are involved in this process, and cellular activities (including cell differentiation) are modulated by hormones, cytokines and growth factors. Within the last 20 years, many factors involved in bone tissue metabolism have been found to be closely related to the inflammatory process. More recently, a cytokine family sharing a common signal transducer (gp130) had been identified, which appears to be a key factor in bone remodelling. This family includes interleukin 6, interleukin 11, oncostatin M, leukaemia inhibitory factor, ciliary neurotrophic factor and cardiotrophin-1. This paper provides an exhaustive review of recent knowledge on the involvement of gp130 cytokine family in bone cell (osteoblast, osteoclast, etc.) differentiation/activation and in osteoarticular pathologies. PMID- 11023661 TI - Integration of the NF-kappaB and mitogen-activated protein kinase/AP-1 pathways at the collagenase-1 promoter: divergence of IL-1 and TNF-dependent signal transduction in rabbit primary synovial fibroblasts. AB - Collagenase-1 (MMP-1) is a protease that is expressed by stromal cells and that is involved in remodeling of the extracellular matrix. IL-1 and TNF-alpha enhance collagenase secretion by stromal cells, and chronic exposure of cells to these cytokines can contribute to connective tissue disease. In this study, we show that the NF-kappaB pathway is required for activation of collagenase-1 transcription in rabbit primary synovial fibroblasts (RSF). Although both IL-1 and TNF activate NF-kappaB in these cells, only IL-1 induces collagenase-1 transcription. We have reported previously that NF-kappaB and AP-1 cooperate to mediate IL-1-induced MMP-1 transcription. Here, we show that IL-1 is superior to TNF at inducing c-Jun synthesis, phosphorylation and binding activity in RSF. Similarly, IL-1 is more effective at activating the mitogen-activated protein kinases (MAPK), including the extracellular signal-regulated kinases (ERK), which are required for IL-1-induced MMP-1 transcription. Thus stimulation of the ERK and AP-1 pathways is an essential component of MMP-1 transcriptional activation, which is deficient in TNF-treated cells. These studies demonstrate cooperation between the MAPK and NF-kappaB signaling pathways for IL-1-dependent collagenase 1 transcription, and they define a dichotomy of IL-1- and TNF-elicited signaling that is relevant to cytokine-mediated connective tissue disease. PMID- 11023663 TI - Soluble CD40 ligand induces beta-chemokine production by macrophages and resistance to HIV-1 entry. AB - CD40 ligand (CD40L) is a cell surface molecule of CD4(+)T cells that interacts with its receptor CD40 on antigen presenting cells to mediate thymus-dependent humoral immunity and inflammatory reactions. We report here that treating monocyte-derived macrophages (MDM) with a trimeric soluble form of CD40L (CD40LT) induced them to secrete high levels of the beta-chemokines RANTES, MIP-1alpha and MIP-1beta that are ligands for CCR5 and able to inhibit HIV-1 entry. CD40LT inhibited the entry of M-tropic HIV-1 reporter viruses. Furthermore, supernatants obtained from CD40LT-stimulated macrophages protected CEMx174-CCR5 cells from infection by HIV-1(JRFL)reporter virus. The inhibitory activity appeared to be due to beta-chemokines present in the supernatant, since pretreating them with a cocktail of antibodies to RANTES, MIP-1alpha and MIP-1beta neutralized the inhibitory activity of the supernatants. In addition, treating monocytes with CD40LT caused CCR5 and CD4 to be downregulated from the cell surface. In vivo, macrophages activated through CD40 could interfere with HIV replication. PMID- 11023662 TI - Interleukin 8, neutrophil-activating peptide-2 and GRO-alpha bind to and elicit cell activation via specific and different amino acid residues of CXCR2. AB - The objective of this investigation was to determine the amino acid residues of the human neutrophil CXC chemokine receptor-2 (CXCR2) that are critical for binding the ligands interleukin 8 (IL-8), neutrophil-activating peptide-2 (NAP 2), and growth-related protein alpha (GROalpha) and critical for receptor mediated signal transduction. Charged residues of the amino terminus and the first extracellular loop of CXCR2 were targeted for point mutagenesis studies. Seven separate CXCR2 mutants (Glu7, Asp9, Glu12, Asp13, Lys108, Asn110, and Lys120, all to Ala) were generated. Based on the Scatchard analysis of radioligand binding studies, the following amino acids were deemed critical for ligand binding: (i) Asp9, Glu12, Lys108, and Lys120 for IL-8 and (ii) Glu7, Asp9, and Glu12 for GROalpha. Point mutations in the amino terminus domain (Asp9 and Glu12) and the first extracellular loop (Lys108, Asn110, and Lys120) of CXCR2 reduced cell activation to all three ligands as measured by changes in intracellular calcium concentration. In conclusion, high-affinity binding of IL 8, NAP-2, and GROalpha to CXCR2 involves interaction with specific and different amino acid residues of CXCR2. Furthermore, we propose that the CXCR2 amino acid residues required for cell activation are not necessarily the same residues required for ligand binding. PMID- 11023664 TI - Route of monocyte differentiation determines their cytokine production profile: CD40 ligation induces interleukin 10 expression. AB - Interleukin 10 is a potent anti-inflammatory and immunomodulatory cytokine. Little is known regarding its induction in monocytes/macrophages, however LPS, a reproducible trigger of IL-10, is augmented by direct contact with T cells. In this context, the role of CD40-ligation is investigated. In the rheumatoid synovium, IL-10 is produced by tissue macrophages. Monocytes primed with M-CSF, a cytokine present in rheumatoid joints, produced IL-1beta, TNF-alpha and IL-10 upon CD40-ligation at an IL-1: TNF-alpha: IL-10 ratio of 10:0.5:1. IFN-gamma primed monocytes, however, predominantly produced TNF-alpha and IL-1beta. Both differentiated monocytes display an endogenous IL-10 activity regulatable by CD40 stimulation. Additionally, these monocytes display differential control by exogenous and endogenous IL-1 and TNF-alpha. M-CSF-primed monocyte IL-10 production was dependent on endogenous TNF-alpha and, to a lesser extent, IL-1, whereas IFN-gamma-primed monocytes were partially dependent on endogenous IL-1. The addition of exogenous IL-1 augments CD40 induced IL-10 production by IFN gamma-primed monocytes. These data indicate that CD40 ligation regulates cell contact mediated macrophage IL-10 and that the route of differentiation determines the cytokine profile. PMID- 11023665 TI - Monocytes are target cells for IL-10 induction by HIV-1 Nef protein. AB - IL-10 plays a pivotal role in the pathogenesis of several diseases and is elevated in sera of HIV-infected patients. Recently, we demonstrated that HIV Nef induces IL-10 mRNA expression as well as IL-10 production using PBMCs, H9 or U937 cells. This induction of IL-10 is inhibited by a calmodulin antagonist, W-7. In the present study, T or B lymphocytes or monocytes were isolated from PBMCs of healthy HIV-negative donors. Production of IL-10 and mRNA gene expression were analyzed on each isolated cell population after treatment with Nef or SEA for 3 24 h. The results show that Nef induces IL-10 production as well as mRNA expression significantly using monocytes but not with T or B lymphocytes. By contrast, SEA induced IL-10 production as well as mRNA expression using T lymphocytes but not with monocytes or B lymphocytes. PMID- 11023666 TI - gp130-Dependent signalling pathway is not enhanced in gp130 transgenic heart after LIF stimulation. AB - Activation of gp130 transduces a hypertrophic signal in the heart, but it is not clear whether signalling through gp130 is enhanced when gp130 is overexpressed in vivo. We generated gp130 transgenic mice (TG) and examined the activation of signalling pathways downstream of gp130 in the hearts. The tyrosine phosphorylation of gp130 was enhanced, the phosphorylation of STAT3 and ERK (extracellular signal regulated kinase) 1/2 was increased and induction of the beta-myosin heavy chain (MHC) gene was observed in TG hearts without significant phenotypic changes. Intravenous administration of leukaemia inhibitory factor (LIF) induced tyrosine phosphorylation of STAT3 and ERK 1/2 and expression of c fos and beta-MHC mRNAs in wild-type littermates' (WT) hearts. However, enhancement of STAT3 and ERK 1/2 phosphorylation or augmented mRNA expressions was not observed in TG hearts after LIF stimulation. Next, STAT-induced STAT inhibitor (SSI) mRNA expression was examined. The expression of SSI-1, SSI-2, and SSI-3 mRNAs was significantly augmented in TG hearts after LIF stimulation. These results indicate that overexpressed gp130 does not always enhance downstream signals in the hearts and suggest that the SSI family plays a role in the regulation of the gp130-dependent signalling pathway in the hearts. PMID- 11023667 TI - Production of a biologically active human interleukin 18 requires its prior synthesis as PRO-IL-18. AB - Interleukin (IL-)18 is an activator of NK cells and a co-inducer of Th(1)cytokines, sharing structural features with the IL-1 family of proteins. Unlike most other cytokines, IL-18 and IL-1beta lack a signal peptide, have an all beta-pleated sheet structure and are synthesized as biologically inactive precursors (pro-IL-18 and pro-IL-1beta). These precursors are cleaved by caspase 1 (IL-1beta-converting enzyme, ICE) to form the biologically active mature cytokines. Direct expression of mature recombinant human IL-18 in E. coli resulted in a partially active cytokine. We tested the possibility that correct folding of huIL-18 requires its prior synthesis as pro-IL-18. Because caspase-1 is not readily available, we constructed an expression vector encoding human pro IL-18 in which the caspase-1 cleavage site was mutated into a factor Xa site. To facilitate purification, the mutated pro-IL-18 cDNA was fused in frame to a glutathione-S-transferase (GST) coding sequence. The GST-pro-IL-18 fusion protein was expressed in E. coli, captured on glutathione agarose and mature human IL-18, exhibiting high biological activity was released upon cleavage with factor Xa. This result indicates that correct folding of huIL-18 occurs at the level of pro IL-18 and provides a practical way to produce biologically active huIL-18. PMID- 11023668 TI - Increased concentrations of plasma IL-18 in patients with hepatic dysfunction after hepatectomy. AB - We investigated the dynamic aspects of circulatory IL-18 and other inflammatory cytokines in patients who underwent a hepatectomy. In patients with post operative hepatic dysfunction, plasma concentrations of these cytokines increased, reflecting severe surgical trauma. IL-6, IL-10 and IFN-gamma increased in the early phase, while IL-18 increased in the later phase after 1 week. Interestingly, the increase in the plasma IL-18 concentration was correlated with that in serum bilirubin levels in hepatectomized patients. Hence, the decrease in the hepatic metabolism of IL-18 may cause the plasma accumulation of IL-18. This mechanism was confirmed using rat experiments. Intravenously administered human IL-18 was excreted into bile. Furthermore, the plasma clearance of human IL-18 was prolonged in bile duct-ligated rats. These results suggest that IL-18 is metabolized in the liver and excreted into bile, and an increase in plasma IL-18 in patients with hepatic dysfunction reflects the decreased metabolism in the liver. PMID- 11023669 TI - Colonic expression and synthesis of interleukin 13 and interleukin 15 in inflammatory bowel disease. AB - A dysregulated local immune reaction with unbalanced cytokine expression seems essential in inflammatory bowel disease (IBD), i.e. ulcerative colitis (UC) and Crohn's disease (CD). Since the roles of interleukin (IL-)13 and IL-15 remain unclear, this study aimed at studying intestinal expression of IL-13 and IL-15 in IBD. METHODS: In colonic biopsies from 24 UC, 18 CD, and 12 controls IL-13 and IL 15 were measured using ELISA, and their gene expressions were assessed by RT-PCR. Leukocytes were visualised histochemically. RESULTS: Concentrations of IL-13 were decreased in UC (median 56 pg/mg tissue; interquartile range 30-99 pg/mg) compared to CD (82 pg/mg tissue; 41-122;P=0.004) and controls (83 pg/mg tissue; 18-134;P>0.05), and lower in active UC (53 pg/mg tissue; 33-96) than in inactive UC (80 pg/mg tissue; 65-99;P=0.02). IL-15 concentrations were higher in CD patients (34 pg/mg tissue; 24-53) as compared to controls (20 pg/mg tissue; 15 21;P=0.001) whilst being 22 pg/mg tissue (15-32) in UC. IL-13 mRNA and IL-15 mRNA were detected in 20% and 15%, respectively. Infiltration of leukocytes correlated inversely with IL-13 levels (P=0.02). CONCLUSION: Active UC is associated with decreased colonic IL-13 suggesting that IL-13 levels are diminished as a part of UC exacerbations, or that exacerbations follow active downregulation of IL-13. PMID- 11023670 TI - Constitutive expression of IL-6-LIKE cytokines in normal bone marrow: implications for pathophysiology of myeloma. AB - Myeloma is a neoplasm thought to "home" to bone marrow. However, evidence for bone-marrow-specific receptors or adhesion molecules expressed on myeloma cells is scanty. Initial myeloma expansion is thought to be due to IL-6 and/or related cytokines. Previous determinations of cytokine expression in bone marrow were performed on bone marrow stromal lines; these findings may not reflect the constitutive pattern of expression in situ. Intracytoplasmic staining for IL-6 like cytokines revealed constitutive expression of some factors in the bone marrow of normal mice, but not spleens. Spleens of myeloma-transplanted SCID mice expressed IL-6-like cytokines, indicative of induction of expression by myeloma. Some cytokines expressed in bone marrow induced myeloma proliferation in the presence of dexamethasone, demonstrating dependence of the myeloma on these cytokines. Our data imply that, rather than "homing" to bone marrow, myeloma cells proliferated within marrow cavities more than in other organs because of growth factors constitutively expressed by bone marrow cells. As myeloma progressed, we observed the induction of growth factor expression in spleen cells. Furthermore, because cytokines other than IL-6 may induce myeloma cell proliferation, therapy aimed at neutralizing IL-6 may not be the most effective method to treat this disease. These findings have implications for both the pathophysiology and therapy of multiple myeloma. PMID- 11023671 TI - Ovine interleukin 12 has biological activity on ovine and human activated peripheral blood mononuclear cells. AB - Interleukin 12 (IL-12) is a heterodimeric cytokine composed of two subunits that form the biologically active p70 molecule, and is a potent inducer of the pro inflammatory cytokine IFN-gamma. In this study the coding sequence for ovine interleukin 12 p35 and p40 subunits was derived by RT-PCR cloning. Ovine p35 and p40 cDNA sequences show a high level of similarity at the nucleic acid and protein levels when compared to corresponding bovine and human sequences. In particular, cysteine residues and N-linked glycosylation sites are conserved between species. Secretion of the IL-12 heterodimer from CHO cells co-transfected with ovine p35 and p40 cDNA was shown by immunoprecipitation of a 60 and 66 kDa protein from transfectant supernatant. In addition, the supernatant from co transfected cells augmented the proliferation of Con A-activated ovine peripheral blood mononuclear cells (PBMNC). Cross-species activity was shown by the enhancement of proliferation of human phytohaemagglutinin (PHA)-activated PBMNC. Supernatants from co-transfectants of hu p35/ov p40 and ov p35/hu p40, to generate chimeric heterodimers, also demonstrated stimulatory activity. Human and chimeric IL-12-induced proliferation of activated PBMNC was inhibited using an anti-human IL-12 polyclonal antibody, however this antibody showed minimal inhibition of ovine IL-12. This study suggests that ovine IL-12 has biological properties similar to its human counterpart. PMID- 11023672 TI - Enhancement of oxidised low-density lipoprotein uptake by macrophages in response to macrophage migration inhibitory factor. AB - We examined the expression of macrophage migration inhibitory factor (MIF) mRNA in murine macrophage cell line (RAW 264.7 cells) in response to oxidized low density lipoprotein (oxLDL), and investigated the influence of MIF on the uptake and degradation of oxLDL by RAW 264.7 cells. MIF mRNA expression was markedly upregulated in the presence of oxLDL. Consistent with this, the MIF level of the culture medium was increased by stimulation with oxLDL in dose- and time dependent manners. Next, we added recombinant rat MIF to the culture medium and examined its effects on the uptake of(125)I-labelled oxLDL. Pretreatment with MIF enhanced both the uptake and degradation of(125)I-oxLDL. Taken together, these results suggest that MIF released from macrophages in response to oxLDL stimulates oxLDL uptake and degradation in an autocrine and paracrine fashion, which potentially results in atherosclerosis. PMID- 11023673 TI - Cytokine regulation of syndecan expression in cells of liver origin. AB - Syndecan-1 and syndecan-2-two cell surface heparan sulfate proteoglycans-were described in normal human liver. Proteoglycans can modulate the effect of cytokines, and cytokines can influence the expression of proteoglycans. In the present work the regulatory effect of IL-1beta, IL-6, TNF-alpha, IFN-gamma and TGF-beta1 on syndecan-1 and syndecan-2 expression of hepatocytes, hepatoma cell lines, liver and skin fibroblasts has been studied. All cytokines were able to influence the steady state level of syndecan-1 and syndecan-2 mRNA. Their action was target cell specific resulting in either up- or downregulation except TGF beta1 that was stimulatory in all cell types examined. PMID- 11023674 TI - Serum levels of ifn-inducible PROTEIN-10 relating to the activity of systemic lupus erythematosus. AB - IFN-inducible protein-10 (IP-10) is supposed to act as a specific chemoattractant for Th(1)cells. Since Th(1)cells and IFN-gamma are shown to be important for developing systemic lupus erythematosus (SLE), we examined the relationship between serum IP-10 levels and the disease activity. Serum IP-10 levels were markedly increased in the SLE patients depending on the level of disease activity, whereas not in the patients with rheumatoid arthritis (RA). On the other hand, serum MCP-1 levels were increased to a similar extent both in RA and inactive SLE patients, and a little more elevated in active SLE patients. Serum IP-10 levels in SLE patients correlated positively and negatively with levels of anti-DNA antibody and complements, respectively, whereas MCP-1 levels correlated less or not at all. These results suggest that serum IP-10 levels could be a good indicator for the activity of SLE and that IP-10 could play an important immunological role in SLE. PMID- 11023675 TI - Keratinocyte growth factor is upregulated by the hyperplasia-inducing drug nifedipine. AB - Keratinocyte growth factor (KGF) is the seventh member of the fibroblast growth factor (FGF) family. It is produced by mesenchymal cells and its activity is specific for epithelial cells, controlling epithelial homeostasis and wound repair in a paracrine manner. Although KGF has been implicated in a number of hyperplastic pathologies, it has not previously been investigated in gingival hyperplasia (GH), an adverse side-effect of three pharmacologically different types of drugs, including the anti-hypertensive drug nifedipine (NIF). The mechanism by which NIF causes GH is not yet known, but we have recently shown that KGF mRNA transcripts are elevated in drug-induced GH in vivo (manuscript submitted). It is therefore possible that the action of NIF is mediated via KGF and, in the present study, using the enzyme-linked immunosorbent assay (ELISA) and the semi-quantitative reverse transcribed-polymerase chain reaction (RT-PCR), we found that NIF upregulates KGF secretion and gene transcription by gingival fibroblasts in vitro. Our results thus suggest that KGF may have an important role in the molecular pathology of GH in vivo. PMID- 11023676 TI - Blood cytokine response of low-dose molgramostim (rhGM-CSF)-treated patients. AB - We examined leukocyte counts and ex vivo cytokine response of whole blood to lipopolysaccharide (LPS) or lipoteichoic acid (LTA) in patients under low-dose molgramostim therapy. Patients were injected subcutaneously daily for ten days with 1 microg/kg (n=9) or 2 microg/kg (n=14) molgramostim. Leukocytosis was observed in all patients, but only the eosinophil fraction was significantly increased in relation to other leukocyte populations. Ex vivo IFN-gamma release was decreased and IL-10 and IL-1ra secretion were increased in response to LPS or LTA. Thus, in non-neutropenic patients, leukocytosis can already be initiated by low doses of molgramostim. The ex vivo cytokine data suggest that these doses prime blood towards a systemic anti-inflammatory response. PMID- 11023677 TI - Cytokines and osteolysis around total hip prostheses. AB - The aim of this work is to assess the correlation between the osteolysis around the prosthesis and the presence of cytokines favouring inflammation in the tissues at the interface between loosened prosthesis and bone. In this study, twenty-nine patients that underwent revision surgery were examined. Bioptic samples were collected at the interface between bone and implant both at the stem and socket level. Semiquantitative immunohistochemistry was performed to detect interleukin 1 alpha, interleukin 1 beta, interleukin 6 and tumour necrosis factor, cytokines that directly cause bone resorption and indirectly induce synthesis of other bone resorbing cytokines. Wear particles were identified and quantified by light microscopy. Radiographic evidence for osteolysis was scored by the Engh and Bobyn score. In tissues collected at the interface, the percentage of cells positive to IL1, IL6 and particularly to TNF increased in relation to the tissues collected at the interface with stable components. The cells occurring in the new capsule do not secrete cytokines in quantities that can be related to severity of wear. Cemented prostheses showed higher incidence of severe osteolysis, and higher levels of cytokines. It can be concluded that TNF, and to a lesser extent IL1 and IL6, are positively related to the severity of osteolysis around the prosthesis and therefore a pharmacological treatment can be hypothesized with anti-inflammatory or anti-cytokine drugs in order to limit or to avoid prosthesis loosening. PMID- 11023678 TI - Oocyte-granulosa cell heterologous gap junctions are required for the coordination of nuclear and cytoplasmic meiotic competence. AB - Homologous gap junctions are generally recognized as a means of coordinating cellular behavior under developmental and homeostatic conditions. In the mammalian ovary, heterologous gap junctions between the oocyte and the granulosa cells have been widely implicated in the regulation of meiotic maturation late in oogenesis. However, the role of oocyte-granulosa cell gap junctions at earlier stages of oogenesis is poorly understood. Stage-specific defects in both oocyte and follicle development have been identified in juvenile mice deficient in heterologous oocyte-granulosa cell gap junctions due to targeted deletion of Gja4, the gene encoding connexin-37. Follicle development arrests at the type 4 preantral stage and although oocytes commence growth, oocyte growth ceases at a diameter of 52 microm (74.3% of control size). Analysis of cell cycle and cytoskeletal markers indicates that oocytes arrest in a G(2) state based on uniform decondensed GV chromatin, interphase microtubule arrays, and nonphosphorylated cytoplasmic centrosomes. Functional assays of meiotic competence confirm that oocytes from connexin-37-deficient mice are unable to enter M phase (initiate meiotic maturation) unless treated with the phosphatase inhibitor okadaic acid (OA). Unlike growing oocytes from heterozygous control animals, OA-treated oocytes from connexin-37-deficient mice respond acutely and progress rapidly to the circular bivalent stage of meiosis I and upon removal from OA rapidly revert to an interphase state. In contrast, OA-treated control incompetent oocytes are slow to respond, exhibit a lower proportion of chromosomal bivalent stage oocytes, but remain in and progress into meiotic M phase upon removal from OA. This study demonstrates that heterologous gap junctional communication is required for the completion of oocyte growth and the acquisition of cytoplasmic meiotic competence. PMID- 11023679 TI - Trk C receptor signaling regulates cardiac myocyte proliferation during early heart development in vivo. AB - Neurotrophin-3 (NT-3) is a member of the neurotrophin family of growth factors, best characterized by its survival- and differentiation-inducing effects on developing neurons bearing the trk C receptor tyrosine kinase. Through analysis of NT-3 and trk C gene-targeted mice we have identified NT-3 as critically regulating cardiac septation, valvulogenesis, and conotruncal formation. Although these defects could reflect cardiac neural crest dysfunction, the expression of NT-3 and trk C by cardiac myocytes prior to neural crest migration prompted analysis of cell-autonomous actions of NT-3 on cardiac myocytes. Retroviral mediated overexpression of truncated trk C receptor lacking kinase activity was used to inhibit activation of trk C by endogenous NT-3, during early heart development in ovo. During the first week of chicken development, expression of truncated trk C reduced myocyte clone size by more than 60% of control clones. Direct mitogenic actions of NT-3 on embryonic cardiac myocytes were demonstrated by analysis of BrdU incorporation or PCNA immunoreactivity in control and truncated trk C-expressing clones. Inhibition of trk C signaling reduced cardiac myocyte proliferation during the first week of development, but had no effect at later times. These studies demonstrate that endogenous NT-3:trk C signaling regulates cardiac myocyte proliferation during cardiac looping and the establishment of ventricular trabeculation but that myocyte proliferation becomes NT-3 independent during the second week of embryogenesis. PMID- 11023680 TI - An overlapping CArG/octamer element is required for regulation of desmin gene transcription in arterial smooth muscle cells. AB - The desmin gene encodes an intermediate filament protein that is present in skeletal, cardiac, and smooth muscle cells. This study shows that the 4-kb upstream region of the murine desmin promoter directs expression of a lacZ reporter gene throughout the heart from E7.5 and in skeletal muscle and vascular smooth muscle cells from E9. 5. The distal fragment (-4005/-2495) is active in arterial smooth muscle cells but not in venous smooth muscle cells or in the heart in vivo. It contains a CArG/octamer overlapping element (designated CArG4) that can bind the serum response factor (SRF) and an Oct-like factor. The desmin distal fragment can replace a SM22alpha regulatory region (-445/-126) that contains two CArG boxes, to cis-activate a minimal (-125/+65) SM22alpha promoter fragment in arterial smooth muscle cells of transgenic embryos. lacZ expression was abolished when mutations were introduced into the desmin CArG4 element that abolished the binding of SRF and/or Oct-like factor. These data suggest that a new type of combined CArG/octamer element plays a prominent role in the regulation of the desmin gene in arterial smooth muscle cells, and SRF and Oct like factor could cooperate to drive specific expression in these cells. PMID- 11023681 TI - A role for E-cadherin in mouse primordial germ cell development. AB - In this study we show that mouse primordial germ cells and fetal germ cells at certain stages of differentiation express E-cadherin and alpha and beta catenins. Moreover, we demonstrate that the formation of germ cell aggregates that rapidly occurs when monodispersed germ cell populations are released from embryonic gonads in culture is E-cadherin mediated, developmentally regulated, and dependent on the sex of the germ cells. Immunoblotting analyses indicate that the lower ability to form aggregates of primordial germ cells in comparison to fetal germ cells is not due to gross changes in E-cadherin expression, altered association with beta catenin, or changes in beta catenin phosphorylation. Investigating possible functions of E-cadherin-mediated adhesion in primordial germ cell development, we found that E-cadherin-mediated adhesion may stimulate the motility of primordial germ cells. Moreover, treatment of primordial germ cells cultured on STO cell monolayers with an anti-E-cadherin antibody caused a significant decrease in their number and markedly reduced their ability to form colonies in vitro. The same in vitro treatment of explanted undifferentiated gonadal ridges cultured for 4 days results in decreased numbers and altered localization of the germ cell inside the gonads. Taken together these results suggest that E-cadherin plays an important role in primordial germ cell migration and homing and may act as a modulator of primordial germ cell development. PMID- 11023682 TI - Programmed cell death in zebrafish rohon beard neurons is influenced by TrkC1/NT 3 signaling. AB - Rohon Beard (RB) cells are embryonic primary sensory neurons that are removed by programmed cell death during larval development in zebrafish. RB somatosensory functions are taken over by neurons of the dorsal root ganglia (DRG), suggesting that RB cell death may be triggered by the differentiation of these ganglia, as has been proposed to be the case in Xenopus. However, here we show that the timing of RB cell death correlates with reduced expression of trkC1, the receptor for neurotrophin NT-3, but not with the appearance of DRG, which differentiate only after most RB cells die. trkC1 is expressed in subpopulations of RB neurons during development, and cell death is initiated only in trkC1-negative neurons, suggesting a role for TrkC1 and its ligand, NT-3, in RB cell survival. In support of this, antibodies that deplete NT-3 induce RB cell death while exogenous application of NT-3 reduces death. In addition, we show that RB cell death can be prevented using a caspase inhibitor, zVADfmk, showing that during normal development, RB cells die by a caspase-dependent programmed cell death pathway possibly triggered by reduced signaling via TrkC1. PMID- 11023683 TI - Spatial distribution and differentiation potential of stem cells in hatchlings and adults in the marine platyhelminth macrostomum sp.: a bromodeoxyuridine analysis. AB - Stem cells (neoblasts) in Platyhelminthes are pluripotent, and likely totipotent, undifferentiated cells which retain throughout adult life the capacity to proliferate and from which all somatic cells as well as the germ cells derive. However, basic data on the pool and heterogeneity of neoblasts, their rates of differentiation into sets and subsets of differentiated cells, and their migration to different body regions are still lacking. To fill this gap, S-phase cells in the macrostomid Macrostomum sp. were labeled with the thymidine analog 5 bromo-2'-deoxyuridine (BrdU). S-phase cells were found to be neoblasts and to be distributed in two bands along the lateral sides of the body leaving unlabeled the median axis of the body and the region anterior to the eyes. This distribution is parallel to that of mitotic cells demonstrated using an antibody to phosphorylated histone H3. At different chase times, clusters of BrdU-labeled cells appear, labeled cells migrate to formerly unlabeled areas, and they differentiate into several somatic cell types and into germ cells. Finally, continuous exposure to BrdU shows an extensive renewal of the epithelial cells. Altogether, these results strengthen the idea of platyhelminth neoblasts as an unparalleled stem-cell system within the Animal Kingdom calling for further investigation. PMID- 11023684 TI - Vibrio fischeri lipopolysaccharide induces developmental apoptosis, but not complete morphogenesis, of the Euprymna scolopes symbiotic light organ. AB - During initiation of the association between the squid host Euprymna scolopes and its bacterial partner Vibrio fischeri, the bacteria induce dramatic morphogenesis of the host symbiotic organ, a portion of which involves the signaling of widespread apoptosis of the cells in a superficial ciliated epithelium on the colonized organ. In this study, we investigated the role in this process of lipopolysaccharide (LPS), a bacterial cell-surface molecule implicated in the induction of animal cell apoptosis in other systems. Purified V. fischeri LPS, as well as the LPS of V. cholerae, Haemophilus influenzae, Escherichia coli, and Shigella flexneri, added in the concentration range of pg/ml to ng/ml, induced apoptosis in epithelial cells 10- to 100-fold above background levels. The absence of species specificity suggested that the conserved lipid A portion of the LPS was the responsible component of the LPS molecule. Lipid A from V. fischeri, E. coli, or S. flexneri induced apoptosis. In addition, strains of H. influenzae carrying a mutation in the htrB gene, which is involved in the synthesis of virulent lipid A, showed a diminished ability to induce apoptosis of host cells. Confocal microscopy using fluorescently labeled LPS indicated that the LPS behaves similar to intact bacterial symbionts, interacting with host cells in the internal crypt spaces and not directly with the superficial epithelium. Although LPS was able to induce apoptosis, it did not induce the full morphogenesis of the ciliated surface, suggesting that multiple signals are necessary to mediate the development of this animal-bacterial mutualism. PMID- 11023686 TI - Genes in the first and fourth inversions of the mouse t complex synergistically mediate sperm capacitation and interactions with the oocyte. AB - The t haplotypes (t) are recent evolutionary derivatives of an alternate form of the mouse t complex region located at the proximal end of chromosome 17. This variant form of approximately 1% of the mouse genome is a source of mutations altering numerous sperm functions crucial for fertilization. Males that carry two t haplotypes (t/t) are invariably sterile. t haplotypes contain four inversions relative to the wild-type t complex (+), so that in matings involving a +/t heterozygote, t is usually transmitted as a single unit. However, rare recombinants have been recovered, which carry only part of the t genotype and express only some of the t-dependent phenotypes. Use of these partial t haplotypes in genetic crosses has resulted in the general location of the two major t male sterility factors, S1 and S2, within inversions 1 and 4, respectively. Since sterility can result from a plethora of sperm defects, we have made a detailed study of various functional parameters of sperm from mice carrying S1 or S2 heterozygously or homozygously or in combination. Both S1 and S2 contain mutations altering sperm functions, including motility, capacitation, binding to the zona pellucida, binding to the oocyte membrane, and penetration of the zona pellucida-free oocyte. Therefore it seems clear that each of these factors contains multiple genes contributing to sterility. Furthermore, our results indicate that genes within S1 interact with genes in S2 for all sperm functions examined. However, S1 and S2 genes affecting motility interact in a purely additive fashion, while S1 and S2 genes affecting most other sperm characteristics interact in a synergistic manner. Additionally, the patterns of synergism between S1 and S2 for abnormalities in capacitation, sperm-oolemma binding, and zona-free oocyte penetration are nearly identical. This suggests that these three defects are caused by mutation of the same gene within each sterility factor. These findings will not only be instrumental in matching the various t haplotype sperm defects to candidate genes for S1 and S2, but will facilitate a more comprehensive understanding of the cellular and genetic mechanisms underlying t haplotype male sterility. PMID- 11023685 TI - brother of rhomboid, a rhomboid-related gene expressed during early Drosophila oogenesis, promotes EGF-R/MAPK signaling. AB - The Drosophila rhomboid (rho) gene participates in localized activation of EGF receptor signaling in various developmental settings. The Rhomboid protein has been proposed to promote presentation and/or processing of the membrane-bound Spitz (mSpi) EGF-related ligand to generate an active diffusible form of the ligand. Here, we report on a new rhomboid-related gene identified by sequence similarity searching that we have named brother of rhomboid (brho). In contrast to rho, which is expressed in complex patterns during many stages of development, brho appears to be expressed only during oogenesis. brho transcripts are present in early oocytes and abut posterior follicle cells which exhibit high levels of MAPK activation. brho, like rho, collaborates with Star to promote signaling through the EGF-R/MAPK pathway, and genetic evidence indicates that Brho can activate both the mSpi and the Grk precursor EGF ligands in the wing. We propose that endogenous brho may activate the oocyte-specific Gurken ligand and thereby participate in defining posterior cell fates in the early follicular epithelium. PMID- 11023687 TI - Pseudomonas pickettii: a common soil and groundwater aerobic bacteria with pathogenic and biodegradation properties. AB - Pseudomonas pickettii is an aerobic, nonfermentative, Gram-negative rod-shaped, bacterium that has been isolated from soil, water, humans, and recently the bovine intestinal tract. It belongs to the rRNA group II of the genus Pseudomonas and has three biovars: Va-1, Va-2, and biovar 3/thomasii. P. pickettii can cause pneumonia, meningitis, endocarditis, and osteomyelitis in humans. It frequently is associated with nosocomial infections that often are linked to contaminated injectable solutions. P. pickettii exhibits remarkable ability to degrade a variety of toxic compounds such as chlorophenols, aromatic hydrocarbons, 2,4 dichlorophenoxyacetic acid, and pentacyclic triterpeniod compounds. The genes that encode for these properties are chromosome- and plasmid-associated. Strains of the organism also have demonstrated resistance to heavy metals, such as cadmium, copper, and zinc. This species can survive in a nutrient-poor environment and use a variety of toxic compounds as carbon and energy sources, making it an ideal candidate for study in the biodegradation of toxic compounds found in wastewater and soils. PMID- 11023688 TI - Influence of cellular density on determination of EC(50) in microalgal growth inhibition tests. AB - Growth inhibition tests for copper were carried out on four marine microalgal species: Chlorella autotrophyca, Nannochloris atomus (Chlorophyceae), Phaeodactylum tricornutum (Bacillariophyceae), and Isochrysis aff. galbana (Primnesiophyceae). The test initial cellular densities were reduced to 50 and 10% from the recommended initial cellular density in most of standardized assays. OECD test protocol (originally described for freshwater) was adapted for seawater. The EC(50) values were reduced when initial cellular density decreased. The green algae used in this study exhibited lower sensitivity than P. tricornutum and quite lower than I. aff. galbana. The latter species was found to be very sensitive to copper. The concept of cellular toxic quote (amount of toxic per cell) is defined in order to improve the results of toxicity tests. PMID- 11023689 TI - Effects of carbofuran, diuron, and nicosulfuron on acetylcholinesterase activity in goldfish (Carassius auratus). AB - Juvenile goldfish (Carassius auratus) were exposed to three widely used pesticides; carbofuran, diuron, and nicosulfuron. Acetylcholinesterase (AChE) activity and molecular forms of AChE were first characterized in brain and skeletal muscle of unexposed fish. Skeletal muscle had higher AChE activity than brain (306 and 215 nmol/min/mg protein, respectively). In brain, four molecular forms of AChE were found: A12, G4, G2, and G1. In the muscle, three molecular forms were found A12, A8, and G2. AChE activity was then evaluated in both tissues of fish exposed to different concentration of pesticides (5, 50, and 500 microg/L) for 6, 12, 24, and 48 h. In brain, AChE activity was significantly inhibited during all the periods of exposure in response to 50 microg/L (19-28%) and 500 microg/L (85-87%) carbofuran. Such effect was observed in the muscle only at 500 microg/L (86-92%). Carbofuran had no effect on the distribution of molecular forms. Significant inhibitions (9-12%) of brain AChE activity were also observed in response to diuron and nicosulfuron at 500 microg/L during all periods of exposure and for 50 microg/L nicosulfuron after 24 and 48 h. This study pointed out short-term effects of exposure to sublethal concentrations of the three pesticides, ranging among different chemical families, on brain and muscle AChE in goldfish. PMID- 11023690 TI - Inhibition of acetylcholinesterase and different ATPases by a novel phosphorothionate (RPR-II) in rat brain. AB - A novel phosphorothionate (2-butenoic acid-3-(diethoxy phosphinothioyl)-methyl ester (RPR-II), synthesized at the Indian Institute of Chemical Technology, Hyderabad, targets its effect on rat brain acetylcholinesterase (AChE) and Na(+) K(+), Mg(2+), and Ca(2+) ATPases, as evident in this investigation. Three subchronic doses 0.014 (low), 0.028 (medium), and 0.042 (high) mg kg(-1) were administered to rats daily for a period of 90 days RPR-II caused statistically significant dose- and time-dependent inhibition in brain AChE and also in Na(+) K(+), Mg(2+), and Ca(2+) ATPases in both male and female rats after 45 and 90 days of treatment. The low dose was generally insignificant while the medium and high doses were significantly effective. Females were more susceptible than males with regard to brain AChE, Na(+)-K(+), and Mg(2+) ATPases, which indicates sexual dimorphism in the treated rats. Interestingly, after 28 days post-treatment, recovery of these enzymes was observed. The relative sensitivities of these enzymes indicated that brain AChE was more sensitive than any of the ATPases, but among the ATPases Na(+)-K(+) ATPase was more susceptible than Ca(2+) or Mg(2+) ATPases. This compound, besides inhibiting the target of organophosphates, AChE, also inhibited different ATPases, suggesting both synaptic transmission and nerve conduction were affected. PMID- 11023691 TI - Effects of sequential exposure to multiple concentrations of methylmercury in the rat hippocampal slice. AB - Two experiments explored the effects of sequential exposure to multiple concentrations of methylmercury (MeHg) on rat hippocampal slice synaptic transmission and excitability in area CA1. When hippocampal slices were exposed to 0.1, 1, 10, and 100 microM MeHg chloride in successive 30-min exposures, MeHg produced an increase in excitability over baseline levels throughout the 1 microM exposure and the first 5 min of the 10 microM exposure, followed by profound suppression of excitability at the 100 microM level. When hippocampal slices were exposed to 10, 25, 50, 75, and 100 microM concentrations, MeHg produced an increase in excitability throughout most of the 10 and 25 microM exposures, followed by profound suppression of excitability at the 50 microM level of exposure. In both series of concentrations, MeHg suppressed local inhibitory systems prior to suppressing excitatory systems. In a third experiment, a single exposure of 50 microM MeHg suppressed both presynaptic and postsynaptic responses recorded in stratum radiatum with the same time course, suggesting that the observed suppressive effects of MeHg were not primarily synaptic. PMID- 11023692 TI - Nonylphenol provokes a vesiculation of the Golgi apparatus in three fish epidermis cultures. AB - The aim of this work was to study the effects of nonylphenol and waste water on the cell ultrastructure of fish skin. Therefore, besides a recently established primary cell culture and a cell line, an epidermal tissue culture of fish was developed and tested. In all three systems a prominent vesiculation of the Golgi apparatus was observed after exposure to nonylphenol, which has not been described before and therefore strongly suggests an effect that might indicate exposure to nonylphenol and/or related substances. The Epithelial papulosum cyprini cell line was the most sensitive to nonylphenol, followed by the primary cell culture of epidermis cells and then the explant tissue culture. The vesiculation of the Golgi apparatus was accompanied by degenerative changes in the two cell cultures only. The lack of degenerative changes in the cells of the tissue culture was discussed with respect to the presence of differentiated cells that might better be able to protect themselves by mucous or by an activated xenobiotic metabolism. In a second type experiment, a waste water sample containing small concentrations of nonylphenol was applied to the cultures. It did not lead to a vesiculation of the Golgi apparatus, probably because the nonylphenol concentrations in the waste water were too low to induce the vesiculation. The cultures exposed to waste water revealed unspecific degenerative cellular changes. Additionally, explant cultures were prepared from fish that had survived a 6-month exposure to polluted river water. In these cultures a higher number of mitochondria containing myelin bodies were observed when compared to control cultures. Consequently, exposure to polluted water containing a mixture of substances in vitro and in vivo was found to lead to degenerative alterations in the ultrastructure of the cells. PMID- 11023693 TI - Chlorotriazine reactive Azo red 120 textile dye induces micronuclei in fish. AB - The main objective of this work was to use micronucleus induction in fish erythrocytes to study the risk to aquatic ecosystems due to the genotoxicity of Chlorotriazine Reactive Azo Red 120 textile dye. The frequencies of micronuclei were studied for three low doses of 1, 5, and 10 mg/L and blood sampling was carried out on the same fish after 3, 6, and 9 days. It was found that micronuclei increased not only in a dose-dependent manner but also in a time dependent way, compared with negative (tap water) and positive (10 ppm benzene) control groups. There was also a slight, time-dependent increase in erythrocyte micronuclei of the control fish specimens. This study proved the genotoxicity of this dye, and suggests that further studies should be made on other dyes and some other toxic industrial pollutant discharges in water ecosystems, using fish as an indicator to monitor pollutant genotoxicity. PMID- 11023694 TI - A battery of toxicity tests as indicators of decontamination in composting oily waste. AB - Heterogeneous oily waste from an old dumping site was composted in three windrows constructed from different proportions of waste, sewage sludge, and bark. The objectives of this pilot study were to examine the usefulness of composting as a treatment method for this particular waste and to study decontamination in the composting process by using a battery of toxicity tests. Five samples from the windrow having intermediate oil concentrations were tested with toxicity tests based on microbes (Pseudomonas putida growth inhibition test, ToxiChromotest, MetPLATE, and three different modifications of a luminescent bacterial test), enzyme inhibition (reverse electron transport), plants (duckweed growth inhibition and red clover seed germination), and soil animals (Folsomia candida, Enchytraeus albidus, and Enchytraeus sp.). The luminescent bacterial tests were used as prescreening tests. Chemical analyses of samples were carried out simultaneously. Both toxicity and oil concentration, including those of polyaromatic hydrocarbons (PAHs), were reduced during composting and soil quality improved significantly. The total oil hydrocarbon concentration decreased from 90,000 to 19,000 mg/kg, measured with the IR method, in 4 months, and from 86,000 to 1400 mg/kg, measured with GC method. The concentration of PAHs decreased from 135 to 23.5 mg/kg. During the fourth month of composting (stabilization stage), the proportion of the heaviest oil fractions (asphaltenes) became dominant. Toxicity varied between different samples and between different bioassays; however, the first sample was significantly more toxic than the others, and most of the tests revealed a decrease in toxicity during the composting process. PMID- 11023695 TI - Persistence and distribution of azinphos-methyl following application to littoral enclosure mesocosms. AB - The organophosphorus insecticide azinphos-methyl was applied once to the surface of 12 of 18 littoral enclosure mesocosms (5x10 m) constructed in a 2-ha pond near Duluth, Minnesota. Water, sediment, macrophytes, and adult fathead minnows were analyzed for residue to determine the persistence, distribution, and mass balance of azinphos-methyl. Nominal treatment concentrations were 0, 0.2, 1, 4, and 20 microg/liter active ingredient. The maximum residue concentration in the water was measured 1h after treatment. The half-life in the water column ranged from 1.2 to 2 days and 95% of the residue dissipated in 5.4 to 10.2 days. Measurable residues were found in the sediment, macrophytes, and fish. Maximum residues in these media were measured at 4, 1, and 0.12 days. respectively. The water and sediment were the most important sorptive compartments for azinphos-methyl residue. The macrophytes and fish were of minor importance, containing only trace amounts of the mass applied. PMID- 11023696 TI - Toxicity of three oil spill remediation techniques to the Australian bass Macquaria novemaculeata. AB - Australian bass, Macquaria novemaculeata, were exposed to the water accommodated fraction (WAF) of Bass Strait crude oil, dispersed crude oil, burnt crude oil, and 4-chlorophenol. The WAF of dispersed crude oil was the most toxic treatment with 96-h LC(50) values of 7. 15% (7.94% upper and 6.42% lower 95% CI) and 7.45% (8.26% upper and 6.71% lower 95% CI). The WAF of crude oil was less toxic, with 96-h LC(50) values of 43.72% (49.21% upper and 38.87% lower 95% CI) and 45.87% (51.51% upper and 40.97% lower 95% CI). The WAF of burnt crude oil was the least toxic treatment with 96-h LC(50) values of 49.81% (63.33% upper and 39.44% lower 95% CI) and 47.28% (59.72% upper and 37.62% lower 95% CI). Sublethal toxicity of the crude oil WAF and burnt crude oil WAF was observed at dilutions seven to eight times less than in the dispersed crude oil WAF. PMID- 11023697 TI - Comparison of four chronic toxicity tests using algae, bacteria, and invertebrates assessed with sixteen chemicals. AB - The performances of four chronic toxicity tests, comprising the Daphnia magna 21 day (d) (crustacean), Brachionus calyciflorus 2-d (rotifer), Pseudokirchneriella subcapitata 72-h (green algae), and the Microtox chronic 22-h (bacteria) tests, were compared. Sixteen chemicals with toxicity covering 6 orders of magnitude were studied. Very high correlations were found between the NOEC/EC(10) Pseudokirchneriella 72-h, NOEC/EC(10) Brachionus 2-d, and the NOEC Daphnia 21-d tests. The toxicological response of rotifers and microalgae were within the same order of magnitude as the response of Daphnia in 80% of cases (13/16 chemicals). The Microtox chronic test also anticipated the overall results of the Daphnia 21 d test, but the prediction was rather imprecise, compared with microalgae and rotifers. The test measuring the algal growth inhibition of P. subcapitata after 72h was the most sensitive bioassay. Toxicity on microalgae after 72h could be estimated after 5h by measuring either the direct fluorescence of either photosynthetic pigments or fluorescein diacetate in 56 and 43% of cases, respectively. The median value of the ratio between EC(10) and EC(50) was 3.75, 2, and 1.5 with the algae, the rotifers, and the bacteria, respectively. PMID- 11023698 TI - Stability constants of metal-humic acid complexes and its role in environmental detoxification. AB - Humic acid with hydroxyl-, phenoxyl-, and carboxyl-reactive groups can form coordination compounds with metals. The ion-exchange equilibrium method using Dowex AG 50W-X8, 20-25 mesh Na(+) form was used to determine stability constants of complexes formed between humic acid (isolated from the soil) 50-250 microg ( approximately 3x10(-5)-15x10(-5) mol/liter) and metal salts solution 200 microg at pH 3.5. The stability constant (log K) for different metal-humic acid complexes indicated the following order of the stabilities of complexes formed between humic acid and metal ions: Cu>Fe>Pb>Ni>Co>Ca>Cd>Zn>Mn>Mg. The data on stability constants demonstrated substantial deviation from Irwing-Williams series reported for divalent ions. The molar humic acid/metal ratios were also calculated. Some of the factors affecting the stability constants such as cation exchange capacity of humus soil, molecular radius, and molecular surface area of humic molecules were also estimated. The significance of the data to predict the behavior of these complexes in the environment is discussed. PMID- 11023699 TI - Changes of soil and plant tissue selenium status in an upland grassland contaminated by selenium-rich agricultural drainage sediment after ten years transformed from a wetland habitat. AB - A field survey was conducted in 1989, 1994, and 1999 in order to monitor the soil and vegetation Se concentrations at the Kesterson upland grassland contaminated by Se-rich drainage sediment. The rate of Se dissipation estimated by the change of soil Se concentration, via volatilization, found to be about 1.1% per year. Soil water-extractable Se increased in 1994, but greatly reduced in 1999. The increase of soil Se concentration in the top 15 cm of soil at the fresh-soil fill sites indicates that the plants were able to effectively take up the soluble soil Se from the lower soil profile and to deposit it on the top of the field. This process may reduce the rate of leaching of soil Se. Overall, the concentration of soil water-soluble Se was relatively low and it is unlikely that problems of transport of Se from the Kesterson soil to the adjacent uncontaminated environment by leaching can occur. Plant tissue Se concentration was found to coincide with the soil water-extractable Se concentration. The average plant tissue Se concentration and soil water-extractable Se detected in 1999 was about 10 microg Se g(-1) and 110 microg Se kg(-1) dry weight, respectively, and the estimated bioaccumulation value of this upland grassland is less than 10% of the previous wetland habitat. Therefore, the existing Kesterson grassland should not be at high risk to the environment. PMID- 11023700 TI - Functional and ultrastructural cell pathology induced by fuel oil in cultured dolphin renal cells. AB - Investigations were undertaken to elucidate in a marine mammal renal cell culture system the toxicity and some of the mechanisms of cytopathology in a standardized preparation following exposure to No. 1 fuel oil. Cell survivability of a cultured SP1K renal cell line from the Atlantic spotted dolphin Stenella plagiodon was reduced in a dose-dependent manner after a 12-h exposure to fuel oil. Early morphologic changes reflecting cytotoxicity, as revealed by transmission electron microscopy, included enlarged rough endoplasmic reticula, cytoplasmic vacuolization, and degenerative cytoplasmic inclusions, but mitochondria remained resistant. Assessment of extracellular proton loss by microphysiometry of cultured cells revealed fuel oil-induced enhancement of proton loss that was dependent upon both protein kinase C and renal epithelial Na(+)/H(+) counter-transport functioning, as the specific inhibitors H-7 and amiloride reduced this stimulatory petroleum effect. Cell cycle progression and apoptosis (programmed cell death) were studied in dolphin renal cells exposed to fuel oil for 12, 24, and 48 hours. The toxicant increased the percentage of cells in GO/GI phase and decreased the percentage of cells in S phase starting after 24 hours. The number of cells undergoing early apoptosis was also increased after 24 hours. PMID- 11023701 TI - Papers to appear in environmental research section A PMID- 11023703 TI - Nitric oxide reactivity and mechanisms involved in its biological effects. AB - Nitric oxide (NO) is implicated in many different biological functions. This is due to its widespread distribution in tissue and to its ability to react with a range of molecules in the organism, of which haemoglobin (Hb), soluble guanylyl cyclase (GC), and superoxide anion are of particular note. In this review we describe the biological pathways of NO and their involvement in its physiological effects and toxicity. This endothelial factor rapidly diffuses into the vascular compartment, and the reaction with the Hb haem group is the main metabolic pathway for endogenous NO. Hb is, therefore, a scavenger for this mediator, which prevents it from reaching the tissue components. NO also reacts with the GC haem group, and this combination is fundamental to its acute vasorelaxing effect. Although molecular oxygen plays a very small part in the oxidization process of NO in biological systems, NO reacts with the superoxide anion to generate peroxynitrite at a rate that is limited only by its diffusion coefficient. This reaction is important in pathological conditions because the peroxynitrite thus formed is a selective oxidant and nitrating agent that interacts with numerous biological molecules, thereby damaging them. In addition, of particular note are the interactions of NO with thiol groups, which may mediate several relevant effects in the organism. NO may also activate endogenous ribosyltransferases, which facilitate the transfer of adenosine diphosphate-ribose groups from nicotine adenine dinucleotide to the G protein amino acid residues. These last two processes may also be involved in the control of arterial tone and more precisely so when chronic NO production takes place. PMID- 11023702 TI - New aspects on the melanocortins and their receptors. AB - Knowledge of melanocortins and their receptors has increased tremendously over the last few years. The cloning of five melanocortin receptors, and the discovery of two endogenous antagonists for these receptors, agouti and agouti-related peptide, have sparked intense interest in the field. Here we give a comprehensive review of the pharmacology, physiology and molecular biology of the melanocortins and their receptors. In particular, we review the roles of the melanocortins in the immune system, behaviour, feeding, the cardiovascular system and melanoma. Moreover, evidence is discussed suggesting that while many of the actions of the melanocortins are mediated via melanocortin receptors, some appear to be mediated via mechanisms distinct from melanocortin receptors. PMID- 11023704 TI - 2-Mercaptoethylamine, radioprotector, inhibits the induction of the oxidative stress-inducible (soi) gene by paraquat in Escherichia coli. AB - To demonstrate the superoxide radical (.O(2)(-)) -scavenging activity of 2 mercaptoethylamine (MEA), we investigated the induction of the oxidative stress inducible (soi) gene fused lacZ gene (soi-28:: lacZ) by the use of paraquat as a source of.O(2)(-). When MEA or cysteine was added to the cultures before paraquat treatment, soi gene induction by paraquat was significantly inhibited. However, a high quantity of ascorbic acid (5 mm) inhibited soi gene induction by paraquat far less than MEA or cysteine did. The induction of soi gene induction by MEA exhibited a dose-dependent manner in the range of over 0.2 mm. The antagonistic molecules on the radioprotective action of MEA, ascorbic acid and cysteine did not counteract MEA action on the inhibition of paraquat-mediated soi gene induction. To clarify that the MEA action on the inhibition of paraquat-mediated soi gene induction may be due, in part, to.O(2)(-)-scavenging activity, we investigated the ability of MEA to inhibit the nitroblue tetrazolium (NBT) reduction mediated by.O(2)(-)generated in the xanthine oxidase/hypoxanthine system in vitro. At concentrations above 1 mm, MEA effectively inhibited the NBT reduction in a concentration-dependent fashion. Our results demonstrated that MEA has an ability to scavenge.O(2)(-), and so protects against.O(2)(-)-mediated damage. PMID- 11023705 TI - Cyclosporine impairs the guanylyl cyclase activity of the natriuretic peptide receptor in the glomerulus. AB - In order to elucidate the involvement of the atrial natriuretic peptide (ANP) and its receptor (natriuretic peptide receptor; NPR) system in cyclosporine-induced nephrotoxicity, we investigated the cyclosporine A (CsA)-induced changes in characteristics of the NPR/guanylyl cyclase system in the glomerulus and inner medulla of the rat kidney. CsA was administered intramuscularly to rats for 2 weeks (CsA group). Particulate guanylyl cyclase activity was measured in glomerular and inner medullary membranes. For receptor characteristics, quantitative in vitro receptor autoradiography was performed. The guanylyl cyclase activity in the glomerulus from the CsA group was attenuated compared with that from the control. However, the activity in the inner medulla was not affected by CsA treatment. Direct application of CsA to normal glomerular membrane completely abolished the ANP-induced guanylyl cyclase activation. Binding studies, using(125)I-ANP, revealed that B(max)was decreased in the CsA group, while K(d)was not affected in the glomerulus. However, in the inner medulla, neither B(max)nor K(d)was affected by CsA treatment. CsA did not displace the(125)I-ANP bindings to NPRs in the normal rat kidney. Local tissue ANP as well as plasma ANP concentration in both groups was not significantly different. These results indicate that CsA impairs the guanylyl cyclase activity mainly in the glomerulus by the decrease in NPR population and/or by direct inhibition, suggesting that the ANP/NPR system might be involved in CsA-induced nephrotoxicity. PMID- 11023706 TI - A deleted form of human hepatocyte growth factor stimulates hepatic lipogenesis and lipoprotein synthesis in rats. AB - Here we report the effect of the recombinant human deleted form of hepatocyte growth factor (dHGF) on lipid metabolism in rats. In primary cultured rat hepatocytes, dHGF accelerated incorporation of [(14)C]acetate into cellular lipids in a concentration-dependent manner. dHGF also increased the gene expression and enzyme activity of glucose-6-phosphate dehydrogenase, a rate limiting enzyme of the pentose phosphate pathway, in hepatocytes. These results suggest that dHGF stimulates hepatocyte lipogenesis through upregulation of the pentose pathway and NADPH formation. Injection of dHGF into normal rats induced elevation of the serum triglyceride, phospholipid and cholesterol levels dose dependently and in the same time course as the liver growth. dHGF injections stimulated the [(14)C]acetate incorporation into the liver lipids, but not into the adipose tissue nor the small intestine. Serum very low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) levels were elevated by dHGF injections. [(14)C]Leucine incorporation into VLDL and LDL was also increased by dHGF injections. In rats with alcohol-induced fatty livers, dHGF treatment markedly diminished the accumulated liver triglyceride, while elevating serum lipid concentrations. The present results indicate that dHGF stimulates exclusively hepatic lipogenesis and increases serum lipoprotein levels in rats. PMID- 11023707 TI - Effects of lidocaine on rabbit isolated thoracic aorta. AB - Lidocaine has been demonstrated to modify both contraction and relaxation of the vascular smooth muscle. Although lidocaine has been shown to inhibit endothelium independent relaxations, the effects of lidocaine on arterial relaxation induced by peroxynitrite, a reaction product of superoxide and nitric oxide, have not been studied. The current study was designed to evaluate the effects of lidocaine on endothelium-dependent and -independent relaxations in isolated rabbit thoracic aorta. Rings of the rabbit thoracic aorta with or without endothelium were mounted for isometric force recording. Concentration-response curves to calcium ionophore A23187 ( 10(-9)to 3 x 10(-6)m), acetylcholine ( 10(-9)to 10(-3)m), sodium nitroprusside (SNP, 10(-9)to 10(-3)m), and peroxynitrite ( 10(-9)to 10( 3)m) were obtained in a cumulative manner. Lidocaine ( 10(-6)to 10(-4)m) was applied 15 min before addition of phenylephrine. Under resting force, lidocaine produced contractions at high concentrations ( 10(-5)to 10(-2)m) in endothelium intact and -denuded arteries but removal of the endothelium did not significantly affect contractile activity. In phenylephrine-precontracted arteries, lidocaine caused concentration-dependent relaxations in both endothelium-intact and denuded arteries. Inhibition of nitric oxide synthase or removal of endothelium did not affect the relaxations to lidocaine. Lidocaine suppressed the endothelium independent relaxations of peroxynitrite, also poly (ADP-ribose) synthetase (PARS) enzyme activator, and SNP at high concentrations. Concentration-dependent vascular relaxations to A23187 and acetylcholine were significantly inhibited by lidocaine. These results suggest that lidocaine can depress vascular relaxations by a complex mechanism including inhibition of PARS enzyme activity. PMID- 11023708 TI - Etoposide encapsulated in positively charged liposomes: pharmacokinetic studies in mice and formulation stability studies. AB - Etoposide is an antineoplastic agent which acts by forming a ternary complex with topoisomerase II and DNA, causing DNA breaks and cell death. In recent studies we have demonstrated that encapsulation in liposomes increases the antitumour efficacy and reduces the adverse effects associated with etoposide. The present study was thus conducted to evaluate whether encapsulation in cationic liposomes altered the pharmacokinetics of etoposide and to study the effect of cholesterol incorporation on the stability of the liposomes. Etoposide-encapsulated unilammellar liposomes were synthesized by thin film hydration followed by extrusion. The drug was administered to Swiss albino mice at a dose of 10 mg kg( 1). The concentration of the drug in plasma was analysed at different time points till 360 min after injection, using a h.p.l.c. method. The terbium chloride dipicolinic acid interaction method was applied to study the stability of the formulation in mouse serum and also following storage at 0( composite function)C over a period of time. The effect of the free and liposomal drug on myelosuppression was evaluated at 10 mg m(-2)and 40 mg m(-2)dose levels by quantifying blood cell counts on day 15 and day 21 following a 5 day course of therapy. Encapsulation in cationic liposomes increased the area under the concentration vs time curve to 42.98 microghml(-1)from 24.18 microghml(-1)in the case of the free drug. Half-life (beta) was 58. 62 and 186 min in the case of free and liposomal etoposide, respectively. In the stability studies, incorporation of cholesterol progressively stabilized the formulation in serum. The use of sucrose at increasing concentrations as a cryoprotectant also increased the shelf stability of the formulation at 0( composite function)C. Toxicity studies using a dose of pure drug revealed that though myelosuppression was evident in both liposomal- and free drug-treated groups on day 15 it was reversed by day 21 following initiation of therapy. The present findings suggest that liposomes could serve as an alternative mode of delivery for etoposide. PMID- 11023709 TI - Development of a physiologically based pharmacokinetic model of organic solvent in rats. AB - A physiologically based pharmacokinetic model of the transfer of organic solvents in rat bodies was developed. The model has six compartments, i.e. lungs, vessel rich tissue, muscles, fat tissue, tail, and liver, each being interconnected by the blood flow system. The transfer of organic solvents was expressed by simultaneous differential equations, which were then solved numerically by a personal computer using a simple spreadsheet program. m -xylene was used to represent organic solvents. The physiological parameters for rats (alveolar ventilation, cardiac output, tissue volume, tissue blood flow, etc.) and physicochemical or biochemical properties (blood/air partition coefficient, tissue/blood partition coefficients, metabolic constants, etc.) of m -xylene were based on the data obtained from the literature and our experiments. The partition coefficient of m -xylene for the tail and the blood flow and the volume of the rat tail were experimentally determined with adult rats. The results of simulation of rat exposure to m -xylene (50 and 500 ppm for 6 h) were essentially in good agreement with the experimental data on rats, i.e. the parent compound (m -xylene) concentration in the tail blood and the cumulative excretion of the metabolites in the urine were consistent. PMID- 11023710 TI - Effects of acetylbergenin against D -galactosamine-induced hepatotoxicity in rats. AB - The hepatoprotective effects of acetylbergenin were examined against D galactosamine (GalN)-induced liver damage in rats, compared with that of bergenin reported previously. Acetylbergenin was synthesized from acetylation of bergenin, isolated from Mallotus japonicus, to increase lipophilic and physiological activities. Acetylbergenin was administered orally once daily for 7 days and then GalN (400 mg kg(-1), i.p.) was injected at 24 h and 96 h after the final administration of acetylbergenin. Acetylbergenin reduced the elevated serum enzyme activities of alanine/aspartate aminotransferase, sorbitol dehydrogenase and gamma -glutamyltransferase and the formation of hepatic malondialdehyde induced by GalN. Acetylbergenin also significantly restored towards normalization the decreased levels of glutathione and the decreased activities of glutathione S transferase and glutathione reductase induced by GalN. Therefore, these results suggest that acetylbergenin has hepatoprotective effects against GalN-induced hepatotoxicity by inhibiting lipid peroxidation and maintaining an adequate level of GSH for the detoxification of xenobiotics as underlying hepatoprotective mechanisms. In addition, lipophilic acetylbergenin showed more activity in the hepatoprotection than that of the much less lipophilic bergenin reported previously. PMID- 11023711 TI - Biological activity of myricetin and its derivatives against human leukemic cell lines in vitro. AB - Two myricetin derivatives, the 3,7, 4('), 5(')- tetramethyl ether of myricetin (1), isolated from the hexane extract of Cistus monspeliensis, and its 3('),5 diacetyl derivative (2) which was synthesized, and the parent compound, myricetin (3), were examined for their in vitro cytotoxic activity against nine human leukemic cell lines, two of which were mdr cell lines. Compound 2 exhibited higher cytostatic and cytotoxic activities in comparison to compound 1, while compound 3 was inactive against all tested cell lines. Vinblastine was used as a control. PMID- 11023712 TI - Synergistic interaction of adrenaline and histamine in human platelet aggregation is mediated through activation of phospholipase, map kinase and cyclo-oxygenase pathways. AB - This study was conducted to examine the mechanism(s) of synergistic interaction of histamine- and adrenaline-mediated human platelet aggregation. We found that platelet aggregation mediated by subthreshold concentrations of histamine (1-4 microm) plus adrenaline (0.5-2 microm) is inhibited by both an alpha(2) adrenoceptor blocker (yohimbine) and a histamine (H1) receptor antagonist (diphenhydramine). In examining the role of the downstream signalling pathway, we found that such an interaction is inhibited by the calcium channel blockers verapamil and diltiazem. However, platelet aggregation by adrenaline plus histamine was inhibited by very low concentrations of the phospholipase C (PLC) inhibitor, U73122 (IC(50)= 1.2 microm), the MEK inhibitor, PD98059 (IC(50)= 1.1 microm) and the cyclo-oxygenase (COX) inhibitor, indomethacin (IC(50)= 7 microm). However the inhibition of receptor tyrosine kinase, protein kinase C and phosphatidylinositol 3-kinase by genistien, chelerythrine and wortmannin, respectively, had no significant effect on aggregation. Similarly the nitric oxide donor (SNAP) had no effect on this synergism. These data suggest that the synergistic effect of histamine and adrenaline during human platelet aggregation is receptor mediated and involves activation of PLC, COX and MAP kinase signalling pathways. PMID- 11023713 TI - Audiological evaluation in adult beta-thalassemia major patients under regular chelation treatment. AB - With the improved life expectation of beta -thalassemia major patients, new clinical problems, such as hearing damage, must be evaluated. Fifty-seven patients (32 F, 25 M; age range 17-32 years) have been studied to define risk factors for development of sensorineural hearing loss. All patients with beta thalassemia major received daily chelation therapy with subcutaneous injection of desferrioxamine (30-50 mgkg(-1)per day). We performed an otological visit and pure tone audiometry as well as impedance; patients were followed for 3 years. Four patients with a conductive hearing loss were excluded; 66.6% had a normal audiogram; 22.8% had a slight sensorineural deficit ( 75 years, (HA-LRTIs 1.7, 1.3-2.2; CA-LRTIs 1.7, 1.0-2.7). Results of multivariable logistic regression analysis showed an increased risk of HA-LRTIs in patients who had a nasogastric tube (3.6, 2.3-3.6), were ventilated (2.3, 1.6-3.2), trauma patients (2.2, 1.5-3.0), chronic obstructive airway disease (COAD), (1.9, 1.5-2.3), a tracheostomy (1.9, 1.3-2.7), prior blood transfusion (1.5, 1.2-1.8), smokers (1.4, 1.1-1.6) or on systemic corticosteroid therapy (OR 1.3, 1.1-1.6). Community acquired LRTIs were positively associated with cystic fibrosis (33.7, 19.1-59.3), HIV (9.8, 6.5-14.8), COAD (4.8, 3.8-4.8), systemic corticosteroid therapy (2.5, 2.2-2.8), tracheostomy (1.8, 1.1-2.9), males (1.2, 1.1-1.3) and smoking (1.2, 1.1 1.4). PMID- 11023719 TI - An outbreak of multidrug-resistant Pseudomonas aeruginosa infection associated with contamination of bronchoscopes and an endoscope washer-disinfector. AB - Over a two-month period, two distinct types of Pseudomonas aeruginosa resistant to ceftazidime and azlocillin were isolated from bronchial specimens of ITU patients who had been previously bronchoscoped. The source of the outbreak was probably a faulty contaminated bronchoscope washer-disinfector which had been purchased a year earlier but not properly maintained. This paper describes the outbreak, the identification and elimination of the source, and the steps taken to prevent recurrence. Several automated, closed washer-disinfectors had been bought by the hospital in response to health and safety concerns about glutaraldehyde disinfection toxicity, but the operation and maintenance of these machines had not been supervised. Several other washer-disinfectors were also found to be faulty. The potential hazards of automated endoscope washer disinfectors and the importance of controlled professional maintenance, servicing and training is discussed. PMID- 11023720 TI - Aspergillus terreus infections in haematological malignancies: molecular epidemiology suggests association with in-hospital plants. AB - During a three-year period nine patients with haematological diseases after myeloablative chemotherapy died from invasive fungal infections caused by Aspergillus terreus. The hospital inanimate environment was monitored and A. terreus was cultured from potted plants in the vicinity of the patients. The patients (N = 14) and the environmental isolates (N = 2) were fingerprinted by RAPD-PCR with four different primers. Based on RAPD patterns the patients' isolates were differentiated into five different types; the environmental isolates represented two types. The isolates of four patients were identical to those found in the environment. Five additional patients were infected by RAPD types not found in the environment. One patient was infected with two different types. The data indicate a hospital-acquired infection in many of the patients and underline the need for careful environmental monitoring of units in which high-risk patients are housed. PMID- 11023721 TI - Impact of hospital-wide surveillance on hospital-acquired infections in an acute care hospital in the Netherlands. AB - The goal of surveillance is to identify hospital-acquired infections (HAI) and risk factors, to apply targeted interventions and to evaluate their effect in an ongoing system. Continuing active surveillance in a 270-bed acute-care hospital is being performed on clinical patients, excluding day-care. The period 1984-1997 is described here. Specific surveillance-based interventions included the introduction of antimicrobial prophylaxis in gynaecology patients with postoperative urinary tract catheters and inpatients scheduled for appendicectomy and hysterectomy. General measures included education, implementation of protocols, feedback of surgeon-specific infection rates. In total, 3545 HAI were found in 13 years of surveillance. The incidence was 4.7/100 admissions and 4. 5/1000 patient days. Age-specific incidences ranged from 1.3 in the age-category 1-14 years, to 10.2 in patients aged 75 years and above. If age-specific incidences had remained at their 1984 level, over 3000 additional infections would have occurred, affecting all age groups except those up to 14 years. The distribution of types of infections differed between services. Following the targeted interventions, the rate of infections in gynaecology decreased from 19.4 per 1000 patient days in 1984 to 2.4 per 1000 patient days in 1996. The rates of wound infection following appendicectomy and hysterectomy decreased by 69% and 82%, respectively, in the period following the institution of antimicrobial prophylaxis. Over 4000 micro-organisms were isolated from the HAI; multi resistant strains were isolated sporadically. We conclude that hospital-wide surveillance of hospital-acquired infections provides appropriate targets for interventions tailored to the specific needs of the hospital. The impact of such interventions can readily be documented from the surveillance data. PMID- 11023723 TI - Evaluation of the cleaning and disinfection efficacy of the DEKO-190; award-based automated washer/disinfector. AB - The DEKO-190 Washer-Disinfector combines both automatic washing and thermal disinfection functions and is designed for the decontamination of ward utensils (such as bedpans and urine bottles) and instruments used in minor surgery prior to sterilization. We undertook a microbiological evaluation of the disinfection efficacy of the machine, using its short wash plus disinfection programme and sealed suspensions of test organisms placed at various points within the instrument. Suspensions of Enterococcus faecalis and Poliovirus were totally inactivated, the counts of aerobic organisms within a stool specimen being reduced by a factor of 10(4)whilst spores of Clostridium perfringens were unaffected. The cleaning efficacy of the DEKO-190 was also evaluated under clinical conditions by visual inspection and was found to be satisfactory. Ward based combined washer-disinfection machines, such as the DEKO-190, have the potential to improve the efficacy of cleaning protocols within healthcare institutions. PMID- 11023722 TI - Effect of delayed infection control measures on a hospital outbreak of methicillin-resistant Staphylococcus aureus. AB - All patients positive for methicillin-resistant Staphylococcus aureus (MRSA) at the University Hospitals of Geneva, Switzerland, between 1989 and 1997 (N = 1771) were included in a cohort study to evaluate the consequences of delayed containment of a hospital-wide outbreak occurring during a 4-year absence of MRSA control measures. The effects of efforts to control both the MRSA reservoir and the number of bacteraemic patients were assessed. Intensive infection control measures were initiated in 1993 and included patient screening, on-site surveillance, contact isolation, a computerized alert system, and hospital-wide promotion of hand hygiene. An increase in the rate of new MRSA-infected or colonized patients was observed between 1989 and 1994 (from 0.05 to 0.60 cases per 100 admissions), which subsequently decreased to 0.24 cases in 1997 (P<0.001). However, the proportion of laboratory-documented methicillin-resistant isolates among all S. aureus showed little variation in the years from 1993 onwards (range, 19-24%), reflecting the result of an increase in the number of screening cultures. The annual number of patients with MRSA bacteraemia strongly correlated with the hospital-wide prevalence of MRSA patients (R(2)= 0.60; P = 0.01) and the rate of new MRSA patients (R(2)= 0.97; P<0.001). Consequently, the attack rate of nosocomial MRSA bacteraemia served as an excellent marker for the MRSA patient reservoir. In conclusion, despite delayed implementation, infection control measures had a substantial impact on both the reservoir of MRSA patients and the attack rate of MRSA bacteraemia. PMID- 11023724 TI - Survival of human coronaviruses 229E and OC43 in suspension and after drying onsurfaces: a possible source ofhospital-acquired infections. AB - Strains OC43 and 229E of human coronaviruses (HCoV) cause one-third of common colds and hospital-acquired upper respiratory tract HCoV infections have been reported in premature newborns. To evaluate possible sources of infection, virus survival was studied in aqueous suspensions and on absorptive and non-absorptive surfaces representative of a hospital environment. Virus susceptibility to chemical disinfection with standard products was also characterized. Virus survived in saline solution for as long as six days but less in culture medium, with or without added cells. After drying, HCoV-229E infectivity was still detectable after 3h on various surfaces (aluminum, sterile latex surgical gloves, sterile sponges) but HCoV-OC43 survived 1h or less. Of the various chemical disinfectants tested, Proviodine reduced the virus infectious titre by at least 50%. This study suggests that surfaces and suspensions can be considered as possible sources of contamination that may lead to hospital-acquired infections with HCoV and should be appropriately disinfected. PMID- 11023725 TI - Transmission of viruses via contact in ahousehold setting: experiments using bacteriophage straight phiX174 as a model virus. AB - Contamination of the environment with pathogens is the prerequisite for contact infections. The aim of this study was to elucidate how viruses can be transmitted from a primary contact person to further individuals. Bacteriophage straight phiX174 was chosen as a model virus. In its stability straight phiX174 is comparable with the most resistant human pathogenic viruses, e.g. polio- or parvoviruses. About 10(7)pfu were applied to exposed contact points such as door handles or the hands of volunteers. After touching of these handles and common social contacts like hand shaking, re-isolation rates were determined from the hands of our test persons. Contaminated door handles and skin surfaces were found to be efficient sources for potential infection. At least 14 persons could be contaminated by horizontal spread, one after the other by touching the same door handle. Successive transmission from one person to another could be followed up to the sixth contact person. These results were confirmed under everyday life conditions in a flat shared by four students. The transmission could not be prevented by the usual standards of hand hygiene, practised in this household. straight phiX174 could be reisolated after 24h from the hands of all persons tested even after normal use and cleaning of their hands. This might be improved by the use of liquid soap dispensers. PMID- 11023726 TI - Could the incidence of postoperative urinary tract infection be reduced by reversing the sequence of vaginal cleansing and urethral catheterization? AB - Postoperative urinary tract infection (UTI) is a common hospital infection after gynaecological operations. A prospective randomized study was performed to examine whether the incidence of UTI could be reduced by reversing the sequence of vaginal cleansing and urethral catheterization. Subjects were randomly allocated to: (1) urethral catheterization before vaginal cleansing; and (2) urethral catheterization after vaginal cleansing. Urine cultures were performed immediately after the procedures as the baseline, on the day of catheter removal, and two days after catheter removal. Patients were examined daily after the operation for any urinary symptoms and fever. The incidences of preoperative asymptomatic bacteriuria were similar in both groups, seven of 84 in group 1 vs. eight of 83 in group 2. Among those with negative urine culture before the operation, 77 in group 1 and 75 in group 2, there was no significant difference in postoperative bacteriuria at catheter removal (23 vs. 22) and two days later (35 vs. 42). No significant difference was noted in the incidence of UTI (5 vs. 10, P = 0.25), with a trend towards less UTI in group 1. Voiding discomfort was more common, and fever less common in group 1. There were significant associations between urine culture results at three different occasions. Bacteriuria at catheter removal is associated with a 7.2 times risk of bacteriuria two days later, 2.4 times risk of urinary symptoms and 3.2 times risk of UTI. Routine surveillance at catheter removal is not cost-effective. We conclude that postoperative bacteriuria and UTI are common. Reversing the sequence of the procedures cannot reduce the incidence. There is no evidence to change the status quo. PMID- 11023727 TI - Evaluation of the bactericidal efficiency of a 2% alkaline glutaraldehyde solution on Mycobacterium xenopi. AB - Mycobacterium xenopi (M. xenopi) has been implicated in hospital-acquired infections associated with colonization of hospital water systems. M. xenopi is considered to be as resistant as other atypical mycobacteria, which are known to be resistant to many disinfecting treatments. However, the efficacy of disinfectants on this organism has not yet been studied. Therefore we decided to evaluate its susceptibility to 2% alkaline glutaraldehyde solution, which is commonly used in hospitals. Tests were conducted using five strains of M. xenopi: three isolated from human samples, an environmental strain and a collection strain. We used a membrane filtration assay and counted surviving bacteria before and after several exposure times (5, 15, 30 and 60 min) with the disinfecting solution. The log10 reduction factor of organisms achieved within 60 min contact ranged from 2.5 to 7.5. This showed M. xenopi to be more resistant to disinfectants than M. tuberculosis or M. smegmatis and suggested that environmental strains may be more resistant to alkaline glutaraldehyde than those isolated from human samples. PMID- 11023728 TI - Transmission of tuberculosis. PMID- 11023729 TI - Infectivity in the blood of mice with a BSE-derived agent. PMID- 11023730 TI - Ralstonia pickettii colonization of patients in an obstetric ward caused by a contaminated irrigation system. PMID- 11023731 TI - Conflicts between a fungal entomopathogen, Zoophthora radicans, and two larval parasitoids of the diamondback moth. AB - Zoophthora radicans (Zygomycetes: Entomophthorales), Diadegma semiclausum (Hymenoptera: Ichneumonidae), and Cotesia plutellae (Hymenoptera: Braconidae) are all natural enemies of the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae). Adult C. plutellae are not susceptible to Z. radicans infection but the pathogen can infect and kill adult D. semiclausum. Infection of adult D. semiclausum prior to exposure to P. xylostella host larvae significantly reduced the number of parasitoid cocoons subsequently developing from the host larvae. Although Z. radicans infection of P. xylostella larvae prior to parasitism by D. semiclausum or C. plutellae always resulted in the death of the immature parasitoids, neither species discriminated between healthy and Z. radicans infected host larvae in an oviposition choice experiment. However, host larvae recently killed by Z. radicans were always rejected by D. semiclausum but sometimes accepted by C. plutellae. At 20 degrees C, egg to pupa development took 6.7 and 7.8 days for D. semiclausum and C. plutellae, respectively. C. plutellae parasitism significantly increased host instar duration but D. semiclausum parasitism did not. Cadavers of P. xylostella larvae parasitized 1 day prior to fungal infection showed no reduction in Z. radicans conidia yield. However, cadavers of larvae parasitized 3 days prior to fungal infection demonstrated a marked decrease in Z. radicans conidia yield. Z. radicans infection of P. xylostella larvae < or = 4 days after parasitism resulted in 100% parasitoid mortality; thereafter, the reduction in parasitoid cocoon yield decreased as the time between parasitism and initiation of fungal infection increased. The extended duration of the host larval stage induced by C. plutellae parasitism increased the availability of the parasitoid to the pathogen. Estimates of interspecific competition indicated a similar pattern for the interaction between Z. radicans and each species of parasitoid. PMID- 11023732 TI - Ultrastructure and cytopathology of a rickettsia-like organism causing systemic infection in the redclaw crayfish, Cherax quadricarinatus (Crustacea: decapoda), in Ecuador. AB - A study of the ultrastructural characteristics of an intracellular bacterium infecting the redclaw crayfish, Cherax quadricarinatus, a pathogen referred to previously as a rickettsia-like organism (RLO), revealed the presence of different developmental stages. These included a rod-shaped and uniformly electron-dense elementary body (EB) and an intermediate body (IB). The length of the EB varied between 0.48 and 0.6 microm, and the diameter was 0.3 microm. The IB was 0.75 to 1.1 microm long by 0.36 to 0.44 microm in diameter. Although the EB of this bacterium has ultrastructural characteristics similar to those of Rickettsiella, no information is available regarding its genetic relationship to this genus, and the intracellular bacterium should continue to be referred to as a rickettsia-like organism. The hemocytes had different levels of infection, and the RLO proliferated inside these cells. The EB appeared to be free in the cytoplasm of infected hemocytes and other cells; however, this might be a fixation artifact. The EB was also contained in membrane-bound vacuoles along with the IB. RLO colonies were observed inside small granular cells. No large granular cells were observed in the sections examined; therefore, no data were obtained regarding infection of this type of hemocyte. The fixed phagocytes on the external side of the terminal hepatic arterioles had an activated interrupted layer containing RLO bacteria. Stem cells in the hematopoietic tissue were also infected, and some cells were apparently being released into circulation. PMID- 11023733 TI - Hemocyte migration during inflammatory-like reaction of Ciona intestinalis (Tunicata, ascidiacea). AB - Following an injuring factor such as the injection of erythrocytes into the tunic of Ciona intestinalis, an inflammatory-like response occurs and blood cells are massively involved in these mechanisms. Electron microscope observations illustrate the infiltration and migration of blood cells throughout the mantle epithelium into the tunic and show several prominent morphological changes. PMID- 11023734 TI - A flow cytometric approach to the study of crustacean cellular immunity. AB - Responses of hemocytes from the crayfish Procambarus zonangulus to stimulation by fungal cell walls (zymosan A) were measured by flow cytometry. Changes in hemocyte physical characteristics were assessed flow cytometrically using forward and side-scatter light parameters, and viability was measured by two-color fluorescent staining with calcein-AM and ethidium homodimer 1. The main effects of zymosan A on crayfish hemocytes were reduction in cell size and viability compared to control mixtures (hemocytes in buffer only). Adding diethyldithiocarbamic acid, an inhibitor of phenoloxidase, to hemocyte and zymosan mixtures delayed the time course of cell size reduction and cell death compared to zymosan-positive controls. The inclusion of trypsin inhibitor in reaction mixtures further delayed the reduction in hemocyte size and cell death, thereby indicating that a proteolytic cascade, along with prophenoloxidase activation, played a key role in generating signal molecules which mediate these cellular responses. In addition to traditional methods such as microscopy and protein chemistry, flow cytometry can provide a simple, reproducible, and sensitive method for evaluating invertebrate hemocyte responses to immunological stimuli. PMID- 11023735 TI - Effect of light energy on alkali-released virions from Anagrapha falcifera nucleopolyhedrovirus. AB - We compared the insecticidal activities of occluded and nonoccluded AfMNPV baculovirus obtained by dissolving the occlusion bodies (OB) with sodium carbonate. Droplet feeding and cotton leaf feeding bioassay techniques were used to determine the dose response against neonate Trichoplusia ni (Hubner) and loss of insecticidal activity when the virus was exposed to simulated sunlight from a xenon light source. Using droplet bioassays to determine a dose response, nonoccluded virus (NOV) was 20 times more active (LC(50) = 4.8 x 10(3) OB/ml, dissolved) than occluded virus (LC(50) = 9.6 x 10(4) OB/ml) when the samples remained wet. However, NOV lost activity when air dried before being tested by droplet (LC(50) > 1.0 x 10(6) OB/ml) or leaf feeding (LC(50) > 3.0 x 10(6) OB/ml) bioassays. Adding sucrose to NOV prevented the loss of insecticidal activity when samples were dried. The activity of NOV with 2% sucrose was similar to that of occluded virus samples, with or without sucrose, in both droplet feeding and leaf feeding assays. These results indicate that the OB protected the insecticidal activity of virions from the detrimental effects of drying. The OB also provided some protection from the detrimental effects of simulated sunlight (xenon) exposure. NOV samples exposed to xenon light had significantly greater loss of insecticidal activity than did similar samples of occluded virus. Without advancement in technologies, such as formulations, possible benefits of increased insecticidal activity from the use of nonoccluded virus is probably not sufficient to offset the rapid loss of activity due to drying or light exposure. PMID- 11023736 TI - Entomophthora muscae resting spore formation in vivo in the host Delia radicum. AB - The formation in vivo of Entomophthora muscae resting spores was investigated in the host, Delia radicum (cabbage root fly), by analysis of field data on the seasonal occurrence of E. muscae resting spores over 4 years. E. muscae resting spores in D. radicum were spherical with an average diameter of 39.4 microm, and the average numbers produced were estimated at 5.7 x 10(4) resting spores/female cadaver. Resting spores were found only after midsummer in D. radicum and almost exclusively in females. The proportion of infected females with resting spores was negatively correlated with average weekly day length after midsummer. We did not detect any significant year effect; thus, the results support the hypothesis that the photoperiod is the most important abiotic factor controlling E. muscae resting spore formation in D. radicum. PMID- 11023737 TI - Characterization of a Bacillus thuringiensis delta-endotoxin which is toxic to insects in three orders. AB - We report here the first Bacillus thuringiensis (Bt) toxin which is toxic to insects from three insect orders (Diptera, Coleoptera, and Lepidoptera). An oligonucleotide probe based on the delta-endotoxin N-terminal sequence was used to detect the gene. A 23-kb BamHI fragment containing the intact gene was identified and cloned from Bt strain YBT-226 plasmid DNA into the vector pBluescript II. Through a series of DNA manipulations the size of this fragment was reduced and the gene sequenced. The deduced amino acid sequence gave a predicted molecular mass of 137 kDa and was identical to a cry1Ba protein from Bt subsp. thuringiensis HD-2, which is now designated as Cry1Ba1 under a new classification scheme. This protein also showed 81.6% similarity with the Cry1B protein (Cry1Bb1) from Bt strain EG 5847. When the YBT-226 cry1Ba1 gene was expressed in an acrystalliferous Bt subsp. israelensis strain it produced irregular bipyramidal crystals during sporulation, which reacted specifically with anti-Cry1Ba antiserum. Bioassays using these crystals after purification resulted in significant mortality at low to moderate concentrations to larvae of the house fly (Musca domestica, Diptera), cottonwood leaf beetle (Chrysomela scripta, Coleoptera), and tobacco hornworm (Manduca sexta, Lepidoptera). This broad-spectrum toxicity was not dependent on presolubilization. In assays with insect cell lines not derived from midgut cells, the soluble toxin killed CH1t (Manduca sexta cells) but was inactive against CF1 (Choristoneura fumiferana cells), Aa(s) (Aedes aegypti), and C2 (Culex quinquefasciatus) mosquito cells. PMID- 11023738 TI - Genetic engineering of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus as an improved pesticide. AB - The Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearNPV) has been registered and is commercially produced in China as a biopesticide to control the bollworm in cotton. However, the virus has a relatively slow speed of action. To improve its efficacy, recombinant HearNPVs were generated by deleting the ecdysteroid UDP-glucosyltransferase (egt) gene (HaCXW1 and HaLM2) or by inserting the insect-specific toxin gene AaIT in the egt locus (HaCXW2) of HearNPV using conventional recombination strategies in insect cell culture. The various recombinants remained genetically stable when cultured in HzAM1 insect cells. Bioassay data showed a significant reduction in the time required for all HearNPV recombinants to kill second instar H. armigera larvae. The LT(50) of the egt deletion recombinants HaCXW1 and HaLM2 was about 27% faster than that of wild type HearNPV. The largest reduction in LT(50) was achieved by inserting the gene for the insect-specific neurotoxin, AaIT, in the egt locus, giving a reduction in LT(50) of 32% compared to wild-type HearNPV. The ability to genetically improve the properties of HearNPV as a biopesticide provides a further opportunity to develop this virus into a commercially viable product to control the bollworm in China. PMID- 11023739 TI - Whole-cell PCR of Bacillus thuringiensis recovered from the phylloplane. PMID- 11023740 TI - Epizootic of the entomopathogenic nematode Steinernema intermedium (Poinar) in an aggregation of the bibionid fly, Bibio marci L. PMID- 11023741 TI - Intraspecific variation in the effects of parasitism by Asobara tabida on phenoloxidase activity of Drosophila melanogaster larvae. PMID- 11023742 TI - Enzymatic activities in European flat oyster, Ostrea edulis, and pacific oyster, Crassostrea gigas, hemolymph. AB - Enzymatic activities in the hemolymph of healthy and Bonamia-infected Ostrea edulis and Crassostrea gigas were studied with a commercial kit for the detection of 19 enzymes: 15 and 16 enzymes, respectively, were detected in the hemolymph of O. edulis and C. gigas and 10 of them showed relatively high activity levels. Most of them existed in both the cell-free fraction of the hemolymph and in the hemocytes. The cell-free hemolymph fraction of Bonamia ostreae-infected European flat oysters showed an elevated enzymatic activity level compared with that of healthy individuals. C. gigas hemocytes possessed higher enzymatic activity levels than O. edulis hemocytes. Differences in enzymatic activities existed in granulocytes and hyalinocytes in both oyster species. The enzyme release from oyster hemocytes seemed to be selective. The infection by B. ostreae induced enzymatic activity variations in European flat oysters. Higher enzyme levels within hemocytes may contribute partly to the natural resistance of C. gigas to the infection by B. ostreae. PMID- 11023743 TI - Effects of long- and short-term passage of insect cells in different culture media on baculovirus replication. AB - Two insect cell lines that had been maintained in both serum-free (SFM) and serum containing (SCM) media for over 5 years were each tested for their ability to replicate baculovirus. The gypsy moth cell line, IPLB-LdEIta (Ld), produced similar (not statistically different) amounts of gypsy moth nucleopolyhedrovirus (LdMNPV) occlusion bodies (OBs) in the two media (serum-free Ex-Cell 400 and TC 100 with 9% (v/v) fetal bovine serum, SCM(1)) but produced more of the Autographa californica nucleopolyhedrovirus (AcMNPV) OBs in SFM than in SCM(1). When Ld cells normally grown in SCM(1) were switched to SFM, production of OBs from both viruses improved and, after three passages, reached higher levels of AcMNPV production than in cells normally maintained in that medium. Alternatively, cells switched from SFM to SCM(1) initially produced as much (in the case of LdMNPV) or higher (in the case of AcMNPV) levels of virus OBs than cells normally maintained in SCM(1) but productivity dropped off over subsequent passages such that after five passages in SCM(1), cells produced substantially fewer OBs of both viruses. A fall armyworm cell line (IPLB-SF21AE; Sf) showed slightly different effects from long- and short-term passage in SFM (Ex-Cell 400) or SCM(2) (TMN-FH). Cells maintained in SFM produced about 20 times more AcMNPV OBs than cells maintained long-term in SCM. Sf cells switched from SFM to SCM maintained the level of production of that seen in SFM at the first passage, but quickly dropped off OB production levels to that normally seen in SCM. Alternatively, SCM-maintained Sf cells produced higher levels at the first passage in SFM and, within five passages in SFM, reached levels found in cells maintained for long term in this medium. Under the conditions in which these two cell lines were infected, the highest levels of AcMNPV OB production in Ld cells were about five times that of Sf cells. In a separate series of experiments, cells normally grown in SFM were passaged over five times in Ex-Cell 400 to which serum was added; both cell lines produced as much virus as that in SFM. These results suggest that it is not the serum per se but rather some other components which differ between the SFM and the SCM formulations that are responsible for the varied virus production obtained in these studies. The results of these studies suggest that a maintenance and virus production protocol can be developed with Ld cells which could improve overall efficiency of virus production. These studies also suggest that long-term maintenance of cells in SFM was not detrimental to their ability to produce baculoviruses. PMID- 11023744 TI - Paenibacillus associated with milky disease in Central and South American scarabs. AB - Thirty-one isolates of bacteria causing milky disease in scarab larvae collected in Central and South America were identified as Paenibacillus popilliae or Paenibacillus lentimorbus by use of DNA similarity analysis. The isolates were more similar to each other than to the North American isolates that are the type strains of the species. All of the bacteria of both species produced parasporal bodies, a characteristic previously believed to be unique to P. popilliae. Screening of the bacteria using PCR with parasporal protein primers revealed differences among the parasporal protein genes of P. popilliae isolates and between the parasporal genes of P. popilliae and P. lentimorbus. In contrast to P. popilliae from North America, none of the isolates from Central and South America was resistant to vancomycin, an indication of an interesting geographic distribution of the resistance genes. PMID- 11023745 TI - Phagocytic activities of the gorgonian coral Swiftia exserta. AB - The cellular response component of body defense in gorgonians and other cnidarians is thought to be carried out by cells with phagocytic capabilities. To test for the phagocytic character of cells, the introduction of foreign particles was employed and observed in both living cells and histological preparations of the gorgonian coral Swiftia exserta. Observations of untreated tissues revealed normal cells and tissue morphologies. A microscopic observation of living cells following the introduction of particles in a cut revealed that only a mixed population of colorless cells phagocytized the particles. Also particles or clumps of particles were seen on the surface of the colorless cells. Subsequent histological observations allowed identity of colorless cells to be inferred as granular amoebocytes, ectodermal cells, and gastrodermal cells. Cells stained for localization of peroxidase (indicative of phagocytic activity) demonstrated the presence of peroxidase-positive cells. Histological preparations revealed that major phagocytosis of particles was associated with tissue trauma. When particles were introduced by means of a cut or inserted thread, phagocytic activity was detected within 2 h. However, it was confined to the granular amoebocytes in the immediate site of trauma. After 24 h, extensive phagocytosis spread throughout a relatively large area surrounding the wound. At that later time, phagocytic cell types included granular amoebocytes, epidermal cells, sclerocytes, mesogleal cells, and gastrodermal cells of the solenia. Observations suggest that trauma induces phagocytosis in cells not normally phagocytic in S. exserta. No localization of phagocytic cells and no mitotic cells were observed at either 2 or 24 h after particle introduction. PMID- 11023746 TI - Effects of incubation temperature on the dose-survival time relationship of Trichoplusia ni larvae infected with Autographa californica nucleopolyhedrovirus. AB - The interaction between virus multiplication and host development was studied by determining the survival time of Trichoplusia ni larvae inoculated with a wide range of doses of Autographa californica nucleopolyhedrovirus and incubated at five different temperatures spanning the biologically relevant range. The results support earlier findings that the course of baculovirus infection follows the mathematical description of the birth-death model. Both in vivo rate of virus increase and larval growth rate increased linearly with increasing temperature from 14 to 29 degrees C; developmental zeros for virus replication and larval growth were estimated from these data to be 10.2 and 10.4 degrees C, respectively. The data were used to generate a description of the combined effects of dose and temperature on median survival time at doses greater than the LD(50). Implications of the lag times before onset of viral replication and between cessation of replication and larval death with respect to model-based estimation of the critical population level (i.e., amount of virus in the host just prior to death) are discussed. PMID- 11023747 TI - Composition and ecological distribution of cry proteins and their genotypes of Bacillus thuringiensis isolates from warehouses in China. AB - The composition and distribution of insecticidal crystal proteins (Cry proteins) and their genotypes of Bacillus thuringiensis isolates from warehouses were evaluated through SDS-PAGE and PCR techniques. The results showed that the electrophoretic patterns of delta-endotoxin crystal preparations were divided into five types. The isolates containing approximately 135 kDa with a 65-kDa protein or only a approximately 135-kDa protein, which amounted to 55.74 and 35.25% of all isolates respectively, were the two major profiles of Cry protein isolated. The distribution of cry genes of B. thuringiensis from warehouses was highly variable. Cry protein genotypes detected in B. thuringiensis isolates included cry1Aa5, cry1Ab9, cry1Ac5, cry1Ba, cry1Ca1, cry1Da1, cry1Ea3, cry2, and cry3 genes, but not cry1Fa2. Among them, cry2, cry1Ac5, and cry1Ab9 genes were the most common in our B. thuringiensis isolates. Most B. thuringiensis isolates contained several cry genes in a total of 18 profiles. Among them, cry1Ac5 with cry1Ea3; cry1Aa5, cry1Ab9, cry1Ac5 with cry1Ea3; and cry1Aa5, cry1Ab9 with cry1Ac5 were the three principal profiles. The distribution of the Cry proteins and cry genes in isolates depended on geography and type of warehouses. Gene profiles may be used as markers for insecticidal activity of B. thuringiensis strains, but they did not directly reflect the toxic level of B. thuringiensis strains. The serotype of B. thuringiensis strains did not directly reflect the specific cry gene profiles in the strains, but certain relationships can be established between the serotype and cry genotype. PMID- 11023748 TI - Characterization and carbon metabolism in fungi pathogenic to Triatoma infestans, a chagas disease vector. AB - The pathogenicity of Metarhizium anisopliae (Ma) and Beauveria bassiana (Bb) isolates against Triatoma infestans, the major vector of Chagas disease in Argentina is reported. A 100% mortality was achieved with mean lethal times varying form 5.8 (Ma6) to 7.7 (Bb5) or 11.1 days (Bb10). The fatty acid, hydrocarbon, and total lipid patterns were compared for glucose-grown and alkane grown Bb10 cultures. The alkane-grown cells showed a lipid pattern different from that of glucose-grown cells, with triacylglyercol as the major lipid fraction, whereas sterols prevailed in the glucose-grown cells. A significant reduction in the relative amounts of linoleic acid diminished the unsaturated/saturated fatty acid ratio for alkane-grown cells; in addition, large amounts of heptacosanoic and eicosanoic acids were detected in the saturated fraction. The hydrocarbon profile of Bb10 showed a saturated chain length distribution,with a marked prevalence for straight chains, ranging from n-C18 to n-C37 in the carbon skeleton, with n-C22 as the major component. Alkane-grown cells showed no qualitative changes in their hydrocarbon fraction, but a similar ratio for odd/even carbon chains. After 48-h incubation assays,[1-(14)C]acetate uptake was largely diminished following a period of alkane growth induction. Glucose-grown cells readily incorporated 19% of the labelinto phospholipids, hydrocarbons, triacylglycerols, and free fatty acids. In contrast, incorporation was reduced to 5.3% for alkane-grown cells, accounting only for phospholipid synthesis. PMID- 11023749 TI - An ultrastructural study of sporidium formation during infection of a rhabditid nematode by large gun cells of Haptoglossa heteromorpha. AB - Recently fired gun cells of Haptoglossa heteromorpha, an aplanosporic nematode parasite, were examined ultrastructurally. The everted tubes of the fired cells had penetrated the cuticle of a nematode, and infective sporidia were developing inside the host body. The nematode cuticle was penetrated by the narrow, walled part of the tube below the needle chamber. The lower unwalled part of the tube tail formed the sporidium. The developing sporidium had a multilayered fibrous outer coating and the plasma membrane was separated from the wall in places. Sporidia contained biphasic membrane-bound vesicles that had been generated by the Golgi dictyosome during gun cell development. Immediately following gun cell firing, the nuclear envelope of the sporidium nucleus was not apparent, and the sporidium nucleus contained clusters of electron-dense particles concentrated in the nucleolar region. We compare the structures and organelles found in the mature gun cell with those in the fired cell and attempt to identify the membranous layers around the sporidium. PMID- 11023750 TI - The marine mites Hyadesia sp. and Copidognathus sp. Associated with the mussel Mytilus galloprovincialis. AB - Two species of marine mites belonging to the families Hyadesiidae and Halacaridae, Hyadesia sp. and Copidognathus sp., respectively, were found associated with the mussel Mytilus galloprovincialis from Baja California in NW Mexico. The first species was found inside the mussel gut with an intensity ranging from one to six mites per mussel and their prevalence was from 20.0 to 46.7%; this species was also found living free in the sediment at a density of 0.7 mite/100 ml. The second species was found on the mantle and gills of the host with an intensity ranging from one to three mites per host and their prevalence was from 3.3 to 6.7%; this species was abundant (4.5 mites/100 ml) and living free in the sediment around mussel clumps. Hyadesia sp. was found alive and attached in the gut of the mussel. A histological analysis revealed this species in the lumen of intestine surrounded by mucus and attached to the epithelial cells of the intestine, where some disorder of epithelial cells was associated. Moreover, this mite may be encapsulated by hemocytes inside the digestive diverticulum, the reproductive follicle, or the connective tissue surrounding the diverticulum. No damages to branches or gills resulting from the presence of Copidognathus sp. were observed. The results suggest that these mites are occasional invaders of mussels; however, as a result of this infestation, Hyadesia sp. may produce damage in the host's tissues. This is the first record of marine mites inside the gut, reproductive follicles, branches, and mantle of a marine bivalve. PMID- 11023751 TI - Intestinal bacteria affect growth of Bacillus thuringiensis in larvae of the oriental tea tortrix, Homona magnanima diakonoff (Lepidoptera: tortricidae). AB - Spores and parasporal crystals of a Bacillus thuringiensis serovar aizawai were fed to fifth instar larvae of the oriental tea tortrix, Homona magnanima, that had been reared aseptically or that had been reared normally. Viable cell numbers of B. thuringiensis and other bacteria in H. magnanima larvae were estimated by homogenization of samples and dilution plating on peptone-polymyxin agar medium for B. thuringiensis cells and on nutrient agar medium for the other bacterial cells. B. thuringiensis did not grow in the larval cadavers of normally reared H. magnanima while bacteria other than B. thuringiensis grew rapidly. In contrast, B. thuringiensis within the larval cadavers of aseptically reared H. magnanima grew and increased 20 times. The bacteria other than B. thuringiensis from the sample homogenates of normally reared larvae that were fed on B. thuringiensis treated diets had the same characteristics as the bacteria isolated from the guts of healthy H. magnanima larvae, which were putatively identified as Streptococcus spp. and Staphylococcus spp., typical intestinal bacteria of insects. The results strongly suggest that intestinal bacteria influence the growth of B. thuringiensis in the larvae. PMID- 11023753 TI - Iridovirus infection in mayfly larvae. PMID- 11023752 TI - Effect of low viability on infectivity of Beauveria bassiana Conidia toward the silverleaf whitefly. PMID- 11023754 TI - Variant Creutzfeldt Jakob Disease. PMID- 11023755 TI - KSHV (HHV8) infection. PMID- 11023756 TI - Current topics in hepatitis B. AB - Over two billion people around the world have been infected with hepatitis B virus, of whom over 350 million are chronic carriers. Some 25% of carriers develop progressive liver disease. The annual mortality from hepatitis B infection and its sequelae is 1-2 million people worldwide.The following current topics are reviewed: immunization strategies against hepatitis B and the kinetics and antibody response; the controversy on screening blood donors for anti-core antibodies; mutations of hepatitis B surface antigen, including evidence that not all such mutants are detectable by current laboratory tests and, finally, the introduction of second generation nucleoside analogues for treatment of chronic hepatitis B infection, including treatment of patients with decompensated liver disease and liver transplantation. PMID- 11023757 TI - Chemoprophylaxis is effective in the prevention of leprosy in endemic countries: a systematic review and meta-analysis. MILEP2 Study Group. Mucosal Immunology of Leprosy. AB - OBJECTIVE: To quantify the efficacy of chemoprophylaxis against leprosy. METHOD: Literature searching of Medline and Embase databases, hand-searching of references and correspondence with investigators. STUDY SELECTION: published papers relating to the prevention of leprosy and the use of chemotherapy in leprosy were identified for critical appraisal. Trials were selected and grouped into three categories according to study design and control groups. DATA ANALYSIS: the relative risks (RR) with 95% confidence intervals were calculated from the original data using a random effects model. To assess the cost effectiveness of chemoprophylaxis, a further analysis of the rates of disease in the trial and control groups was done based on the numbers needed to be treated (NNT) to prevent one new case of leprosy. RESULTS: A total of 14 trials were identified from 127 published papers on chemoprophylaxis of leprosy. The trials were categorized into randomized controlled trials, non-randomized controlled trials, and uncontrolled trials. The overall results of the meta-analysis shows that chemoprophylaxis gives around 60% protection against leprosy. The NNT are low in trials of household contacts. CONCLUSIONS: The evidence shows that chemoprophylaxis against leprosy is an effective way to reduce the incidence of leprosy, particularly in household contacts. The role of chemoprophylaxis needs to be re-examined using newer drugs given the continuing case detection rates globally. PMID- 11023758 TI - Disseminated Mycobacterium simiae infection in patients with AIDS. AB - OBJECTIVES: To report our experience with disseminated Mycobacterium simiae disease in patients with AIDS, and review other cases reported in the literature. METHODS: We retrospectively reviewed all cases of M. simiae that were isolated from sterile body sites over a 9-year period at the University Health System Hospital at San Antonio, Texas, U.S.A. Data included patient demographics, clinical features, other accompanying opportunistic infections, in vitro susceptibility, therapy and outcome. RESULTS: Ten cases of M. simiae disseminated disease were identified. All of them were inpatients with AIDS. Another nine cases of disseminated infection in AIDS patients were reported in the literature. Advanced AIDS with absolute CD4 counts of less than 50 and an associated AIDS defining illness characterized all cases. Persistent fever and debilitation without localizing signs were the most common clinical features. Our patients responded poorly to antimycobacterial drugs and died within 6 months of diagnosis. The only reported successful therapy was in patients who responded well to highly active antiretroviral therapy and antimycobacterial regimens containing clarithromycin, ethambutol and ciprofloxacin. CONCLUSIONS: Clinical presentation of M. simiae infection mimics Mycobacterium avium complex, with fever and progressive debilitation, but is less responsive to therapy. Immuno reconstitution with potent antiretroviral therapy may be the best therapy for such resistant disease. PMID- 11023759 TI - Treatment of external genital warts in men with imiquimod 2% in cream. A placebo controlled, double-blind study. AB - OBJECTIVE: The purpose of this double-blind, placebo-controlled study was to evaluate the safety, clinical efficacy and tolerability of imiquimod (2%) in cream to cure external genital warts in males. METHODS: Preselected male patients (n=60) ranging between 18 and 50 years of age (mean 24.2) harbouring 558 lesions (mean 9.3) with clinical, histopathological and polymerase chain reaction (PCR) confirmed diagnosis of human papilloma virus (HPV) infection were randomized to two parallel groups. Each patient was allocated a precoded 25g tube, and instructions on how to apply the trial medication to their lesions at home once daily for three consecutive days per week (max. 12 application in 4 weeks). To evaluate the safety, clinical efficacy and tolerance, patients were exa-mined on a weekly basis. Cure was defined as the total elimination of treated warts with PCR, and Southern blot hybridization confirmed negative HPV DNA. RESULTS: By the end of the treatment, 40% (24/60) patients and 49.8% (278/558) warts were cured. Breaking the code revealed that imiquimod cream had cured 70% (21/30) patients and 86.8% of warts, while placebo healed three subjects and 28 warts (P=0.0001). Eleven patients (18.3%), predominantly in the imiquimod cream group, experienced mild to moderate, non-objective, drug-related side effects with no dropouts. The study was followed up for 18 months from the first day of the treatment, and among the 26 cured patients, one in the imiquimod cream group and two in the placebo had a relapse after 14 months. CONCLUSION: The study demonstrated that 2% imiquimod in cream with mild non-objective side effects is safe, tolerable and significantly more effective than placebo in curing external genital warts in males. PMID- 11023760 TI - Epidemiology of acute viral respiratory tract infections in Korean children. AB - OBJECTIVE: Viruses are the most common causes of respiratory tract infection in children. We investigated the aetiologies and the epidemiological features of acute viral respiratory tract infections in Korean children. METHODS: We tried to isolate respiratory syncytial virus (RSV) and parainfluenza virus from January 1994, influenza virus from February 1995, and adenovirus from April 1996 through August 1998, and identified the isolated viruses by indirect immunofluorescence (IF) staining in the children hospitalized with acute respiratory tract infections (ARTI). RESULTS: Virus was identified in 360 of 1389 (25.9%) nasopharyngeal aspirates cultured. Of a total of 392 viruses, 164 (41.8%) RSV, 90 (23%) parainfluenza virus, 66 (16.8%) influenza A virus, 54 (13.8%) adenovirus, and 18 (4.6%) influenza B virus were cultured, including cases in mixed viral infections. The male to female ratio of the culture-positive patients was 2:1, and the proportions of the patients aged >6 months, 6-11 months, 1, 2, 3, 4, 5, 6 7, 8-9, and >10 years were 22.5, 29.5, 25.7, 9.5, 3.8, 3.8, 1.7, 1.7, 1.2, and 0.6%, respectively. The major clinical diagnosis was bronchiolitis for RSV, croup for parainfluenza virus, and pneumonia for adenovirus and influenza virus. Infections by RSV, parainfluenza virus, and influenza virus occurred in annual epidemics, and infections by adenovirus occurred annually with or without epidemics. There were somewhat larger epidemics by adenovirus and influenza virus in May to July 1996 and March to June 1997, respectively. CONCLUSIONS: Viral agents are one of the main aetiologies and the main causes of admission in Korean children with ARTI. PMID- 11023761 TI - Presenting clinical features and C-reactive protein in the prediction of a positive stool culture in patients with diarrhoea. AB - OBJECTIVES: To devise a scoring system by which clinical features and C-reactive protein (CRP) can be used to predict a positive stool culture in patients admitted with acute diarrhoea. METHODS: One hundred and thirty-two patients admitted to the Regional Infection Unit with diarrhoea thought to be due to bacterial gastroenteritis were included. Clinical features, CRP and outcome of stool culture were recorded, together with the final diagnosis. RESULTS: Forty one patients had bacterial gastroenteritis characterized by the isolation of a bacterial enteropath (BGE). Sixty-three patients had non-specific gastroenteritis, defined as more than three loose stools per day with no bacterial enteropath isolated (NSGE). In 28 patients another diagnosis was made (Others). More of the patients with BGE (91%) had abdominal pain as compared with those with NSGE (67%) and Others (61%) (P=0.01). The mean duration of symptoms was longer in the Others group (6.14 days) as compared with patients with BGE (3.29) and NSGE (3.25) (P=0.01). The mean CRP was significantly higher in those with BGE (113.9mg/l) and Others (116.9mg/l) as compared to the NSGE patients (38.9mg/l) (P=0.001). A scoring system was devised which incorporated the presence or absence of abdominal pain (+10 or 0), the duration of symptoms (-10, for 5 or more days, 0 for less than 5 days of symptoms) and the CRP (CRP<50=0, CRP>50=5). A score of 15 or more predicted 79% of patients with BGE, while a score of <15 predicted 87% of those with NSGE and 86% of those with another diagnosis. CONCLUSIONS: This simple scoring system may be useful in predicting the positivity of stool culture, and therefore may be helpful in targeting those small number of patients who require antimicrobial therapy after hospital admission. We would not, however, favour reliance on this scoring system alone to choose whom to treat with antimicrobials. PMID- 11023762 TI - Species-specific prevalence of vaginal candidiasis among patients with diabetes mellitus and its relation to their glycaemic status. AB - OBJECTIVES: Non- C. albicans Candida species are increasingly being recognized as the cause of vulvo-vaginal candidiasis. These species are often less susceptible to antifungal agents. Patients with diabetes mellitus are at risk for vulvo vaginal candidasis. We assessed the species-specific prevalence rate and risk of candidiasis in patients with diabetes mellitus and healthy controls. METHODS: Genital tract examination, direct microscopy and fungal cultures of discharge collected by high vaginal swab were undertaken among 78 consecutive patients with diabetes mellitus (mean (+/-sd) age 32+/-12 years and body mass index (BMI) 22.3+/-5.5kg/m(2)) and 88 age- and BMI-matched healthy females. Glycaemic control in the diabetic cohort was assessed by measuring total glycosylated haemoglobin. RESULTS: Candida species were isolated in 36 of 78 (46%) subjects with diabetes mellitus and in 21 of 88 (23%) healthy subjects (Chi-squared 9.11, P=0.0025). The predominant Candida species isolated in diabetics with vulvo-vaginal candidiasis were Candida glabrata (39%), C. albicans (26%) and C. tropicalis (17%). In contrast, in the control group, C. albicans, C. glabrata and C. hemulonii comprised 30% each, with none having C. tropicalis infection (for C. tropicalis: diabetic vs. control; 17% vs. nil, P=0. 05). Among the diabetic group, subjects with vulvo-vaginal candidiasis had significantly higher mean HbA1 when compared to those who had no such infection (12.8+/-2.6% vs. 9.7+/-1.7% respectively, P=0.001). The overall accuracy of direct microscopy and clinical examination for predicting vulvo-vaginal candidiasis was only 77% and 51%, respectively, in the diabetic group, and 83% and 65% in the control group. CONCLUSIONS: Patients with diabetes mellitus had a high prevalence rate (46%) of vulvo-vaginal candidiasis with relative risk of 2.45. The non- C. albicans species such as C. glabrata and C. tropicalis were the predominant species isolated among them. There seems to be a significant link between hyperglycaemia and vulvo-vaginal candidiasis. PMID- 11023763 TI - Tuberculosis-associated severe CD4+ T-lymphocytopenia in HIV-seronegative patients from Dakar. SIDAK Research Group. AB - OBJECTIVES: To determine the frequency and associated features of severe CD4+ T lymphocytopenia (<300 cells/mm(3)) in HIV-seronegative patients with tuberculosis. METHODS: Statistical analysis of 430 consecutively enrolled HIV seronegative inpatients with tuberculosis in two teaching hospitals in Dakar, Senegal. RESULTS: The mean CD4 + cell count was 602+/-318.3 cells/mm(3). CD4 + cell counts were below 300 cells/mm(3)in 62 patients (14.4%). Patients with fewer than 300 CD4+ cells/mm(3)differed from those with higher counts in being less likely to have a positive smear for acid-fast bacilli; in having a higher frequency of extrapulmonary involvement (pleural effusion, adenopathy and miliary disease) and oral candidiasis; and in having smaller tuberculin reactions, lower haemoglobin levels, less cavitation and less patchy infiltration. After adjustment for gender and age, all differences remained except miliary disease. CONCLUSIONS: A substantial percentage (14.4%) of HIV-seronegative hospitalized patients for tuberculosis in a West African country presented with severe CD4 + T lymphocyte depletion and had clinical and radiographic features indicative of more advanced disease and accompanying immunodepression. These results and those already published suggest that tuberculosis should be regarded as one of the diseases associated with a subgroup of patients with "idiopathic CD4 + T lymphocytopenia". PMID- 11023764 TI - General non-specific morbidity is reduced after vaccination within the third month of life--the Greifswald study. AB - OBJECTIVES: The incidence of many serious infectious diseases fundamentally decline as a success of consequent vaccination regimens. However, it is a matter of discussion if vaccination might cause unspecific negative side effects on the immune system. To answer this, we performed a clinical study on children with the question as to whether there is an enhanced frequency of infection diseases after vaccination or not. METHODS: The study population (n=496) was randomized to a group of vaccinated children (first vaccination on the 60th day of life, n=201) and a group of unvaccinated children (first vaccination on the 90th day of life, n=295). Frequencies of unspecific, morbidity-related signs were recorded by the mothers with a diary card. These data were taken for further statistical analysis to determine if the factor "vaccination" does have a significant effect on the variable "morbidity". RESULTS: Various infectious disease-associated symptoms (vomiting, coughing, signs of rhinitis, restlessness, rash and pain) were significantly less often seen in vaccinated than in non-vaccinated children. CONCLUSIONS: Our study revealed that children who received vaccination against diphtheria, pertussis, tetanus, HiB and poliomyelitis simultaneously within the third month of life do not exhibit enhanced frequencies of infectious disease associated symptoms. In contrary, the frequencies of infection-associated symptoms were found to be significantly reduced. This might be caused by a vaccination-associated unspecific enhancement of immunological activity (e.g. mediated by interleukin 2) or by other presently still unknown factors. PMID- 11023765 TI - Medical treatment of a pacemaker endocarditis due to Candida albicans and to Candida glabrata. AB - We describe a case of pacemaker infection due to two fungal species: Candida albicans and C. glabrata. Transthoracic echocardiography showed a large vegetation on the intraventricular wires. Because of severe underlying diseases, surgery was believed to be contraindicated. The patient was treated using high dose of fluconazole, resulting in clinical improvement and negative blood cultures. However, 2 months later, the patient underwent a fatal stroke. At autopsy, a large vegetation was found only all along the wires. Postmortem culture of the infected material was positive for both C. albicans and C. glabrata. PMID- 11023766 TI - Ventriculoperitoneal shunt infection by Candida glabrata in an adult. AB - We describe the first case of infection of a neurosurgical shunt by Candida glabrata in an adult. The risk factors, clinical picture and response to therapy have been similar to similar cases caused by other Candida sp. We must emphasize the unequivocal features of infection and the rapid and probably complete response to removal of the device. PMID- 11023767 TI - Fatal prosthetic valve endocarditis due to Campylobacter fetus. AB - Campylobacter fetus subspecies fetus has a predilection for vascular endothelium. This report describes a case of a prosthetic valve infective endocarditis due to this microorganism. Despite adequate antibiotic combination treatment, valvular vegetations and perivalvular suppurative complications evolved, leading to valvular dysfunction, heart failure and death. Only three other cases of C. fetus prosthetic valve endocarditis have been described previously. Campylobacter fetus bacteraemia must be considered cautiously, because of the risk of late cardiac or vascular complications. PMID- 11023768 TI - Progressive multifocal leukoencephalopathy treated with cidofovir in HIV-infected patients receiving highly active anti-retroviral therapy. AB - Progressive multifocal leukoencephalopathy (PML) has been a devastating disease for HIV-1-infected patients since the beginning of the AIDS pandemic. Currently, highly active antiretroviral therapy (HAART) seems to improve the outcome of PML in many patients. We describe two cases of PML whose course worsened in spite of HAART, but had a good response to cidofovir. PMID- 11023769 TI - Comparison of fitness of two isolates of Mycobacterium tuberculosis, one of which had developed multi-drug resistance during the course of treatment. AB - OBJECTIVES: We report the cases of two patients, brother and sister, both with pulmonary tuberculosis. Both patients complied poorly with treatment. One developed multi-drug resistant disease, whilst the other did not. We aimed to show that the two infecting strains were the same, and then to compare the fitness of the resistant strain to that of the sensitive strain. METHODS: The isolates were typed by RFLP. The fitness of the multi-drug resistant tuberculosis strain was determined by calculating the ratio of generation produced by the drug resistant and a drug-susceptible strain in a mixed culture. The number of bacteria present in this broth culture was estimated using the Miles and Misra technique. The number of drug-resistant bacteria present was determined by inoculating aliquots of broth onto Middlebrook 7H10 agar with 5mg/l rifampicin. RESULTS: The infecting strain of Mycobacterium tuberculosis was shown to be the same on RFLP typing in both cases. It was found that the multi-drug resistant organism had decreased fitness compared to the sensitive organism. CONCLUSION: The decreased relative fitness of the resistant strain implies a physiologic cal cost for the development of drug resistance. PMID- 11023770 TI - Clinical rubella reinfection during pregnancy in a previously vaccinated woman. AB - We report a documented case of clinically apparent rubella reinfection during pregnancy with rubelliform rash and fever followed by lymphodenopathy at the 18th week of gestation, in a previously vaccinated woman with haemagglutination inhibition (HI) antibody titre of 1:32. The serological tests results (including neutralizing antibodies) demonstrated a significant rise in her rubella specific IgG level with strongly positive IgM reactivity. In addition, rubella-specific IgG antibody avidity testing displayed high avidity index (53-88%) typical of rubella reinfection. Umbilical cord blood, drawn by sonographic-guided cordocentesis at 24 weeks' gestation, was found to be negative for rubella specific IgM antibody. The pregnancy was continued to term, and a healthy infant was born. PMID- 11023771 TI - Acute meningococcaemia complicated by late onset gastrointestinal vasculitis. AB - Extra-meningeal and non-cutaneous manifestations of meningococcal infection are well recognized but rare. Herein we report a little recognized complication of meningococcaemia, namely late-onset gastrointestinal vasculitis. In the case presented, the prostacycline analogue iloprost was used in an attempt to minimize incipient digital gangrene ina patient with evidence of protracted immunological phenomena. In addition, diclofenac was used to treat severe arthralgia. Withdrawal of iloprost on day 19 was associated with abdominal pain and peritonism. Laparotomy demonstrated ischaemia and ulceration in the caecum, and histology revealed full thickness mucosal ulceration with a prominent vasculitic process. It is postulated that these findings were immunologically mediated, with possible rebound vasoconstriction following withdrawal of iloprost. It is also possible that NSAID use contributed to the findings via cyto-toxicity to mucosal cells. PMID- 11023772 TI - Hepatitis B exacerbation with a precore mutant virus following withdrawal of lamivudine in a human immunodeficiency virus-infected patient. AB - Chronic active hepatitis B exacerbations have been reported following development of resistance to or withdrawal of lamivudine in HIV-infected patients. A 38-year old woman with HIV and chronic HBV infections was hospitalized because of acute hepatitis. The occurrence of cytolysis with replication of HBV 2 months after withdrawing lamivudine suggests that our patient experienced a severe reactivation of HBV infection due to the modification of her treatment. Sequencing of the HBV precore region showed the strain to be a mutant. We conclude that lamivudine should not be stopped in HIV- and HBV-infected patients, but could be continued at the dose of 100mg/day as used in isolated HBV infection. PMID- 11023773 TI - Mycobacterium avium complex cervical lymphadenitis in an immunocompetent adult patient. PMID- 11023774 TI - Cryptococcal infection in an HIV-positive Ugandan population. PMID- 11023775 TI - Abdominal abscess due to leuconostoc species in a liver transplant recipient. PMID- 11023776 TI - The prevalence of hepatitis B infection in adults with no recognized increased risk of infection. PMID- 11023777 TI - PiOTEYSigma AB - In this issue we visit the association between climate and infection, and between what we eat or drink and what we get as a consequence. Copyright 2000 The British Infection Society. PMID- 11023778 TI - What's new in HIV AB - The management of co-infection of HIV with tuberculosis is a massive worldwide problem, and as the epidemiology of HIV changes in the U.K. , one that is being increasingly confronted in this country. Of considerable relevance, therefore, is the expert panel discussion included in this bulletin. Also, David Moore summarizes the more interesting and important presentations on opportunistic infections from the recent XIII International AIDS Conference held in Durban, and Nicola Smith provides a clear synopsis of HIV-2 infection, from its epidemiology to the management and treatment.S. Morris-Jones and M.R. Nelson Copyright 2000 The British Infection Society. PMID- 11023779 TI - Alteration of a single tryptophan residue of the cellulose-binding domain blocks secretion of the Erwinia chrysanthemi Cel5 cellulase (ex-EGZ) via the type II system. AB - Cel5 (formerly known as endoglucanase Z) of Erwinia chrysanthemi is secreted by the Out type II pathway. Previous studies have shown that the catalytic domain (CD), linker region (LR) and cellulose-binding domain (CBD) each contain information needed for secretion. The aim of this work was to further investigate the secretion-related information present in the CBD(Cel5). Firstly(, )deleting a surface-exposed flexible loop had no effect on secretion. This indicated that some structural freedom is tolerated by the type II system. Secondly, mutation of a single tryptophan residue, previously shown to be important for binding to cellulose, i.e. Trp43, was found also to impair secretion. This indicated that the flat cellulose-binding surface of CBD(Cel5 )contains secretion-related information. Thirdly, CBD(Cel5) was substituted by the CBD(EGG) of Alteromonas haloplanctis endoglucanase G, yielding a hybrid protein CD(Cel5)-LR(Cel5) CBD(EGG) that exhibited 90 % identity with Cel5, including the Trp43 residue. The hybrid protein was not secreted. This indicated that the Trp43 residue is necessary but not sufficient for secretion. Here we propose a model in which the secretion of Cel5 involves a transient intramolecular interaction between the cellulose-binding surface of CBD(Cel5) and a region close to the entry into the active site in CD(Cel5). Once secreted, the protein may then open out to allow the cellulose-binding surface of CBD(Cel5 )to interact with the surface of the cellulose substrate. An implication of this model is that protein molecules fold to a specific secretion-competent conformation prior to secretion that is different from the folding state of the secreted species. PMID- 11023780 TI - Increasing the thermostability of staphylococcal nuclease: implications for the origin of protein thermostability. AB - Seven hyper-stable multiple mutants have been constructed in staphylococcal nuclease by various combinations of eight different stabilizing single mutants. The stabilities of these multiple mutants determined by guanidine hydrochloride denaturation were 3.4 to 5.6 kcal/mol higher than that of the wild-type. Their thermal denaturation midpoint temperatures were 12.6 to 22.9 deg. C higher than that of the wild-type. These are among the greatest increases in protein stability and thermal denaturation midpoint temperature relative to the wild-type yet attained. There has been great interest in understanding how proteins found in thermophilic organisms are stabilized. One frequently cited theory is that the packing of hydrophobic side-chains is improved in the cores of proteins isolated from thermophiles when compared to proteins from mesophiles. The crystal structures of four single and five multiple stabilizing mutants of staphylococcal nuclease were solved to high resolution. No large overall structural change was found, with most changes localized around the sites of mutation. Rearrangements were observed in the packing of side-chains in the major hydrophobic core, although none of the mutations was in the core. It is surprising that detailed structural analysis showed that packing had improved, with the volume of the mutant protein's hydrophobic cores decreasing as protein stability increased. Further, the number of van der Waals interactions in the entire protein showed an experimentally significant increase correlated with increasing stability. These results indicate that optimization of packing follows as a natural consequence of increased protein thermostability and that good packing is not necessarily the proximate cause of high stability. Another popular theory is that thermostable proteins have more electrostatic and hydrogen bonding interactions and these are responsible for the high stabilities. The mutants here show that increased numbers of electrostatic and hydrogen bonding interactions are not obligatory for large increases in protein stability. PMID- 11023781 TI - The essential function of Not1 lies within the Ccr4-Not complex. AB - The five Saccharomyces cerevisiae Not proteins are associated with the Ccr4 and Caf1 proteins in 1.2 MDa and 2 MDa complexes. The Not proteins have been proposed to repress transcription of promoters that do not contain a canonical TATA sequence, while the Ccr4 and Caf1 proteins are required for non-fermentative gene expression. The mechanism of transcriptional regulation by the Ccr4-Not complex is unknown and the role of its different components is unclear. Only Not1p is essential for yeast viability.Here, we show that most strains carrying combinations of two null alleles of the non-essential CCR4-NOT genes are non viable. This would suggest that the Ccr4-Not complex is essential. We find that Not1p consists of at least two domains, a C-terminal domain that is essential for yeast viability, and a N-terminal domain that is dispensable but required for yeast wild-type growth. The essential C-terminal domain of Not1p can associate with Not5p, and both proteins are present in 1.2 and 2 MDa complexes in the absence of the N-terminal Not1p domain. In contrast, in the absence of the N terminal domain of Not1p, Ccr4p does not efficiently associate in large complexes nor with the C-terminal domain of Not1p. Healthy growth is observed when both domains of Not1p are expressed in trans, and is correlated with their physical association, together with Ccr4p, in large complexes. These results are consistent with the essential function of Not1p lying within the Ccr4-Not complex. PMID- 11023782 TI - Substrate recognition and induced DNA deformation by transposase at the target capture stage of Tn10 transposition. AB - The bacterial transposon Tn10 inserts preferentially into sites that conform to a 9 bp consensus sequence: 5' NGCTNAGCN 3'. However, this sequence is not on its own sufficient to confer target specificity as the base-pairs flanking this sequence also contribute significantly to target-site selection. We have performed a series of "contact-probing experiments" to define directly the protein-DNA interactions that govern target-site selection in the Tn10 system. The HisG1 hotspot for Tn10 insertion was the main focus here. We infer that there is a rather broad zone ( approximately 24 bp) of contact between transposase and target DNA in the target-capture complex. This includes base-specific contacts at all of the purine residues in the consensus positions of the target core and primarily backbone contacts out to 7-8 bp in the two flanking regions immediately adjacent to the core. Also, highly localized sites of chemical hypersensitivity are identified that reveal symmetrically disposed deformations in DNA structure in the target-capture complex. Furthermore, the level of strand transfer is shown to be reduced by phosphorothioate substitution of phosphate groups at or close to the sites of target DNA deformation. Interestingly, for one particular target DNA, a mutant form of HisG1 called MutF, the above phosphorothioate inhibition of strand transfer is suppressed by replacing Mg(2+) with Mn(2+). Based on these results a model for sequence-specific target capture is proposed which attempts to define possible relationships between transposase interactions with the target core and flanking sequences, transposase-induced DNA deformation of the target site and divalent metal ion binding to the target-capture complex. PMID- 11023783 TI - Ligand-induced conformational change in the minimized insulin receptor. AB - Within the class of insulin and insulin-like growth factor receptors, detailed information about the molecular recognition event at the hormone-receptor interface is limited by the absence of suitable co-crystals. We describe the use of a biologically active insulin derivative labeled with the NBD fluorophore (B29NBD-insulin) to characterize the mechanism of reversible 1:1 complex formation with a fragment of the insulin receptor ectodomain. The accompanying 40 % increase in the fluorescence quantum yield of the label provides the basis for a dynamic study of the hormone-receptor binding event. Stopped-flow fluorescence experiments show that the kinetics of complex formation are biphasic comprising a bimolecular binding event followed by a conformational change. Displacement with excess unlabeled insulin gave monophasic kinetics of dissociation. The rate data are rationalized in terms of available experiments on mutant receptors and the X ray structure of a non-binding fragment of the receptor of the homologous insulin like growth factor (IGF-1). PMID- 11023784 TI - Polymerization, three-dimensional structure and mechanical properties of Ddictyostelium versus rabbit muscle actin filaments. AB - To assess more systematically functional differences among non-muscle and muscle actins and the effect of specific mutations on their function, we compared actin from Dictyostelium discoideum (D-actin) with actin from rabbit skeletal muscle (R actin) with respect to the formation of filaments, their three-dimensional structure and mechanical properties. With Mg(2+) occupying the single high affinity divalent cation-binding site, the course of polymerization is very similar for the two types of actin. In contrast, when Ca(2+ )is bound, D-actin exhibits a significantly longer lag phase at the onset of polymerization than R actin. Crossover spacing and helical screw angle of negatively stained filaments are similar for D and R-F-actin filaments, irrespective of the tightly bound divalent cation. However, three-dimensional helical reconstructions reveal that the intersubunit contacts along the two long-pitch helical strands of D-(Ca)F actin filaments are more tenuous compared to those in R-(Ca)F-actin filaments. D (Mg)F-actin filaments on the other hand exhibit more massive contacts between the two long-pitch helical strands than R-(Mg)F-actin filaments. Moreover, in contrast to the structure of R-F-actin filaments which is not significantly modulated by the divalent cation, the intersubunit contacts both along and between the two long-pitch helical strands are weaker in D-(Ca)F-actin compared to D-(Mg)F-actin filaments. Consistent with these structural differences, D-(Ca)F actin filaments were significantly more flexible than D-(Mg)F-actin. Taken together, this work documents that despite being highly conserved, muscle and non muscle actins exhibit subtle differences in terms of their polymerization behavior, and the three-dimensional structure and mechanical properties of their F-actin filaments which, in turn, may account for their functional diversity. PMID- 11023785 TI - Electron microscopic evidence for the assembly of soluble pentameric extracellular domains of the nicotinic acetylcholine receptor. AB - Exploitation of soluble extracellular domains (ECDs) of the nicotinic acetylcholine receptor may provide a route to crystallographic studies aimed at exploring the structure and function of the intact receptor. The first step towards this goal is to manufacture and isolate soluble fragments that fold and assemble to form a functionally relevant complex. The baculovirus insect cell expression system was used to co-express soluble ECDs of all four muscle-type nicotinic acetylcholine receptor subunits (alpha, beta, gamma & delta-ECD) from Torpedo. Protein complexes were purified using either the conformationally sensitive monoclonal antibody mAb35, specific for a folded alpha subunit, or a NiNTA affinity resin, specific for a polyhistidine tag engineered on the delta ECD. Western blotting with subunit specific antibodies confirmed the co expression of each ECD and furthermore, indicated that the alpha, beta and gamma ECDs were being co-purified with the polyhistidine-tagged delta-ECD. Chemical cross-linking was used to show that these co-purified proteins had indeed interacted specifically to form soluble oligomeric complexes. A low-resolution, three-dimensional image of these purified complexes, composed only of ECDs, was obtained by electron microscopy. They were shown to resemble the extracellular vestibule of the native receptor, having the same pseudo-pentameric symmetry, size and shape. Expression of incomplete sets of the four nicotinic acetylcholine receptor ECDs did not yield detectable complexes. PMID- 11023786 TI - Structure of native and expanded sobemoviruses by electron cryo-microscopy and image reconstruction. AB - Rice yellow mottle virus (RYMV) and southern bean mosaic virus, cowpea strain (SCPMV) are members of the Sobemovirus genus of RNA-containing viruses. We used electron cryo-microscopy (cryo-EM) and icosahedral image analysis to examine the native structures of these two viruses at 25 A resolution. Both viruses have a single tightly packed capsid layer with 180 subunits assembled on a T=3 icosahedral lattice. Distinctive crown-like pentamers emanate from the 12 5-fold axes of symmetry. The exterior face of SCPMV displays deep valleys along the 2 fold axes and protrusions at the quasi-3-fold axes. While having a similar topography, the surface of RYMV is comparatively smooth. Two concentric shells of density reside beneath the capsid layer of RYMV and SCPMV, which we interpret as ordered regions of genomic RNA. In the presence of divalent cations, SCPMV particles swell and fracture, whereas the expanded form of RYMV is stable. We previously proposed that the cell-to-cell movement of RYMV in xylem involves chelation of Ca(2+) from pit membranes of infected cells, thereby stabilizing the capsid shells and allowing a pathway for spread of RYMV through destabilized membranes. In the context of this model, we propose that the expanded form of RYMV is an intermediate in the in vivo assembly of virions. PMID- 11023787 TI - Towards understanding a molecular switch mechanism: thermodynamic and crystallographic studies of the signal transduction protein CheY. AB - The signal transduction protein CheY displays an alpha/beta-parallel polypeptide folding, including a highly unstable helix alpha4 and a strongly charged active site. Helix alpha4 has been shown to adopt various positions and conformations in different crystal structures, suggesting that it is a mobile segment. Furthermore, the instability of this helix is believed to have functional significance because it is involved in protein-protein contacts with the transmitter protein kinase CheA, the target protein FliM and the phosphatase CheZ. The active site of CheY comprises a cluster of three aspartic acid residues and a lysine residue, all of which participate in the binding of the Mg(2+) needed for the protein activation. Two steps were followed to study the activation mechanism of CheY upon phosphorylation: first, we independently substituted the three aspartic acid residues in the active site with alanine; second, several mutations were designed in helix alpha 4, both to increase its level of stability and to improve its packing against the protein core. The structural and thermodynamic analysis of these mutant proteins provides further evidence of the connection between the active-site area and helix alpha 4, and helps to understand how small movements at the active site are transmitted and amplified to the protein surface. PMID- 11023788 TI - Solution structure of the two N-terminal RNA-binding domains of nucleolin and NMR study of the interaction with its RNA target. AB - Nucleolin is an abundant 70 kDa nucleolar protein involved in many aspects of ribosomal RNA biogenesis. The central region of nucleolin contains four tandem consensus RNA-binding domains (RBD). The two most N-terminal domains (RBD12) bind with nanomolar affinity to an RNA stem-loop containing the consensus sequence UCCCGA in the loop. We have determined the solution structure of nucleolin RBD12 in its free form and have studied its interaction with a 22 nt RNA stem-loop using multidimensional NMR spectroscopy. The two RBDs adopt the expected beta alpha beta beta alpha beta fold, but the position of the beta 2 strand in both domains differs from what was predicted from sequence alignments. RBD1 and RBD2 are significantly different from each others and this is likely important in their sequence specific recognition of the RNA. RBD1 has a longer alpha-helix 1 and a shorter beta 2-beta 3 loop than RBD2, and differs from most other RBDs in these respects. The two RBDs are separated by a 12 amino acid flexible linker and do not interact with one another in the free protein. This linker becomes ordered when RBD12 binds to the RNA. Analysis of the observed NOEs between the protein and the RNA indicates that both RBDs interact with the RNA loop via their beta sheet. Each domain binds residues on one side of the loop; specifically, RBD2 contacts the 5' side and RBD1 contacts the 3'. PMID- 11023789 TI - Solution structure of the TAZ2 (CH3) domain of the transcriptional adaptor protein CBP. AB - The TAZ2 (CH3) domain of the transcriptional adapter protein CBP has been implicated in direct functional interactions with numerous cellular transcription factors and viral oncoproteins. The solution structure of the TAZ2 domain of murine CBP has been determined by nuclear magnetic resonance (NMR). The protein adopts a novel helical fold stabilized by three zinc ions, each of which is bound to one histidine and three cysteine ligands in HCCC-type motifs. Each zinc binding site is formed from the carboxy terminus of an alpha-helix, a short loop, and the amino terminus of the next alpha-helix. A peptide derived from the N terminal transactivation domain of p53 binds specifically to one face of the TAZ2 domain. The close similarities between the TAZ2 and TAZ1 (CH1 domain of CBP/p300) sequences suggest that both domains will adopt similar three-dimensional structures. PMID- 11023790 TI - Transferrins: iron release from lactoferrin. AB - Iron loss in vitro by the iron scavenger bovine lactoferrin was investigated in acidic media in the presence of three different monoanions (NO(3)(-), Cl(-) and Br(-)) and one dianion (SO(4)(2-)). Holo and monoferric C-site lactoferrins lose iron in acidic media (pH< or =3.5) by a four-step mechanism. The first two steps describe modifications in the conformation affecting the whole protein, which occur also with apolactoferrin. These two processes are independent of iron load and are followed by a third step consisting of the gain of two protons. This third step is kinetically controlled by the interaction with two Cl(-), Br(-) and NO(3)(-) or one SO(4)(2-). In the fourth step, iron loss is under the kinetic control of a slow gain of two protons; third-order rate-constants k(2), 4.3(+/ 0.2)x10(3), 3.4(+/-0.5)x10(3), 3.3(+/-0.5)x10(3) and 1.5(+/-0.5)x10(3) M(-2) s( 1) when the protein is in interaction with SO(4)(2-), NO(3)(-), Cl(-) or Br(-), respectively. This step is accompanied by the loss of the interaction with the anions; equilibrium constant K(2), 20+/-5 mM, 1.0(+/-0.2)x10(-1), 1.5(+/-0.5)x10( 1) and 1.0(+/-0.3)x10(-1) M(2), for SO(4)(-), NO(3)(-), Cl(-) and Br(-), respectively. This mechanism is very different from that determined in mildly acidic media at low ionic strength (micro<0.5) for the iron transport proteins, serum transferrin and ovotransferrin, with which no prior change in conformation or interaction with anions is required. These differences may result from the fact that in the transport proteins, the interdomain hydrogen bonds that consolidate the closed conformation of the iron-binding cleft occur between amino acid side-chain residues that can protonate in mildly acidic media. With bovine lactoferrin, most of the interdomain hydrogen bonds involved in the C-site and one of those involved in the N-site occur between amino acid side-chain residues that cannot protonate. The breaking of the interdomain H-bond upon protonation can trigger the opening of the iron cleft, facilitating iron loss in serum transferrin and ovotransferrin. This situation is, however, different in lactoferrin, where iron loss requires a prior change in conformation. This can explain why lactoferrin does not lose its iron load in acidic media and why it is not involved in iron transport in acidic endosomes. PMID- 11023791 TI - Dissecting the hydrogen exchange properties of insulin under amyloid fibril forming conditions: a site-specific investigation by mass spectrometry. AB - We have examined the hydrogen exchange properties of bovine insulin under solution conditions that cause it to aggregate and eventually form amyloid fibrils. The results have been obtained at the residue-specific level using peptic digestion and mass spectrometry. A total of 19 peptides were assigned to regions of the protein and their exchange properties monitored for a period of 24 hours. The results of the peptic digestion show that residues A13 to A21 and B11 to B30 are more susceptible to proteolysis than the N-terminal regions of the protein. A total of 15 slowly exchanging amides were observed for insulin under these solution conditions. Location of the protected amides was carried out using a peptic-digestion protocol at low pH. Chromatographic separation was not required. This enabled a direct comparison of the peptides within the same mass spectrum. From kinetic analysis of the rates slow exchange has been located to 4(+/-1) backbone amides in the A13-A19 helix and 6(+/-1) in the B chain helix. The remaining 5(+/-1) are assigned to helix A2-A8. Taken together the results from digestion and hydrogen exchange show that at low pH and relatively high concentrations the C termini of both chains are susceptible to proteolysis but that the solution structure contains the native state helices. More generally the results demonstrate that mass spectrometry can be applied to study site-specific hydrogen exchange properties of proteins even under conditions where they are known to be partially folded and aggregate extensively in solution. PMID- 11023792 TI - Identification of an allosteric anion-binding site on O-acetylserine sulfhydrylase: structure of the enzyme with chloride bound. AB - A new crystal structure of O-acetylserine sulfhydrylase (OASS) has been solved with chloride bound at an allosteric site and sulfate bound at the active site. The bound anions result in a new "inhibited" conformation, that differs from the "open" native or "closed" external aldimine conformations. The allosteric site is located at the OASS dimer interface. The new inhibited structure involves a change in the position of the "moveable domain" (residues 87-131) to a location that differs from that in the open or closed forms. Formation of the external aldimine with substrate is stabilized by interaction of the alpha-carboxyl group of the substrate with a substrate-binding loop that is part of the moveable domain. The inhibited conformation prevents the substrate-binding loop from interacting with the alpha-carboxyl group, and hinders formation of the external Schiff base and thus subsequent chemistry. Chloride may be an analog of sulfide, the physiological inhibitor. Finally, these results suggest that OASS represents a new class of PLP-dependent enzymes that is regulated by small anions. PMID- 11023793 TI - Correlating amino acid conservation with function in tyrosyl-tRNA synthetase. AB - Sequence comparisons have been combined with mutational and kinetic analyses to elucidate how the catalytic mechanism of Bacillus stearothermophilus tyrosyl-tRNA synthetase evolved. Catalysis of tRNA(Tyr) aminoacylation by tyrosyl-tRNA synthetase involves two steps: activation of the tyrosine substrate by ATP to form an enzyme-bound tyrosyl-adenylate intermediate, and transfer of tyrosine from the tyrosyl-adenylate intermediate to tRNA(Tyr). Previous investigations indicate that the class I conserved KMSKS motif is involved in only the first step of the reaction (i.e. tyrosine activation). Here, we demonstrate that the class I conserved HIGH motif also is involved only in the tyrosine activation step. In contrast, one amino acid that is conserved in a subset of the class I aminoacyl-tRNA synthetases, Thr40, and two amino acids that are present only in tyrosyl-tRNA synthetases, Lys82 and Arg86, stabilize the transition states for both steps of the tRNA aminoacylation reaction. These results imply that stabilization of the transition state for the first step of the reaction by the class I aminoacyl-tRNA synthetases preceded stabilization of the transition state for the second step of the reaction. This is consistent with the hypothesis that the ability of aminoacyl-tRNA synthetases to catalyze the activation of amino acids with ATP preceded their ability to catalyze attachment of the amino acid to the 3' end of tRNA. We propose that the primordial aminoacyl-tRNA synthetases replaced a ribozyme whose function was to promote the reaction of amino acids and other small molecules with ATP. PMID- 11023794 TI - Stabilization of the transition state for the transfer of tyrosine to tRNA(Tyr) by tyrosyl-tRNA synthetase. AB - Aminoacylation of tRNA(Tyr) involves two steps: (1) tyrosine activation to form the tyrosyl-adenylate intermediate; and (2) transfer of tyrosine from the tyrosyl adenylate intermediate to tRNA(Tyr). In Bacillus stearothermophilus tyrosyl-tRNA synthetase, Asp78, Tyr169, and Gln173 have been shown to form hydrogen bonds with the alpha-ammonium group of the tyrosine substrate during the first step of the aminoacylation reaction. Asp194 and Gln195 stabilize the transition state complex for the first step of the reaction by hydrogen bonding with the 2'-hydroxyl group of AMP and the carboxylate oxygen atom of tyrosine, respectively. Here, the roles that Asp78, Tyr169, Gln173, Asp194, and Gln195 play in catalysis of the second step of the reaction are investigated. Pre-steady-state kinetic analyses of alanine variants at each of these positions shows that while the replacement of Gln173 by alanine does not affect the initial binding of the tRNA(Tyr) substrate, it destabilizes the transition state complex for the second step of the reaction by 2.3 kcal/mol. None of the other alanine substitutions affects either the initial binding of the tRNA(Tyr) substrate or the stability of the transition state for the second step of the aminoacylation reaction. Taken together, the results presented here and the accompanying paper are consistent with a concerted reaction mechanism for the transfer of tyrosine to tRNA(Tyr), and suggest that catalysis of the second step of tRNA(Tyr) aminoacylation involves stabilization of a transition state in which the scissile acylphosphate bond of the tyrosyl adenylate species is strained. Cleavage of the scissile bond on the breakdown of the transition state alleviates this strain. PMID- 11023795 TI - Recognition of structurally diverse substrates by type II 3-hydroxyacyl-CoA dehydrogenase (HADH II)/amyloid-beta binding alcohol dehydrogenase (ABAD). AB - Human type II hydroxyacyl-CoA dehydrogenase/amyloid-beta binding alcohol dehydrogenase (HADH II/ABAD) is an oxidoreductase whose salient features include broad substrate specificity, encompassing 3-hydroxyacyl-CoA derivatives, hydroxysteroids, alcohols and beta-hydroxybutyrate, and the capacity to bind amyloid-beta peptide, leading to propagation of amyloid-induced cell stress. In this study, we examine the structure and enzymatic activity of the homologous rat HADH II/ABAD enzyme. We report the crystal structure of rat HADH II/ABAD as a binary complex with its NADH cofactor to 2.0 A resolution, as a ternary complex with NAD(+) and 3-ketobutyrate (acetoacetate) to 1.4 A resolution, and as a ternary complex with NADH and 17 beta-estradiol to 1.7 A resolution. This first crystal structure of an HADH II confirms these enzymes are closely related to the short-chain hydroxysteroid dehydrogenases and differ substantially from the classic, type I 3-hydroxyacyl-CoA dehydrogenases. Binding of the ketobutyrate substrate is accompanied by closure of the active site specificity loop, whereas the steroid substrate does not appear to require closure for binding. Despite the different chemical nature of the two bound substrates, the presentation of chemical groups within the active site of each complex is remarkably similar, allowing a general mechanism for catalytic activity to be proposed. There is a characteristic extension to the active site that is likely to accommodate the CoA moiety of 3-hydroxyacyl-CoA substrates. Rat HADH II/ABAD also binds amyloid-beta (1-40) peptide with a K(D) of 21 nM, which is similar to the interaction exhibited between this peptide and human HADH II/ABAD. These studies provide the first structural insights into HADH II/ABAD interaction with its substrates, and indicate the relevance of the rodent enzyme and associated rodent models for analysis of HADH II/ABAD's physiologic and pathophysiologic properties. PMID- 11023796 TI - Comparison of three methyl-coenzyme M reductases from phylogenetically distant organisms: unusual amino acid modification, conservation and adaptation. AB - The nickel enzyme methyl-coenzyme M reductase (MCR) catalyzes the terminal step of methane formation in the energy metabolism of all methanogenic archaea. In this reaction methyl-coenzyme M and coenzyme B are converted to methane and the heterodisulfide of coenzyme M and coenzyme B. The crystal structures of methyl coenzyme M reductase from Methanosarcina barkeri (growth temperature optimum, 37 degrees C) and Methanopyrus kandleri (growth temperature optimum, 98 degrees C) were determined and compared with the known structure of MCR from Methanobacterium thermoautotrophicum (growth temperature optimum, 65 degrees C). The active sites of MCR from M. barkeri and M. kandleri were almost identical to that of M. thermoautotrophicum and predominantly occupied by coenzyme M and coenzyme B. The electron density at 1.6 A resolution of the M. barkeri enzyme revealed that four of the five modified amino acid residues of MCR from M. thermoautotrophicum, namely a thiopeptide, an S-methylcysteine, a 1-N methylhistidine and a 5-methylarginine were also present. Analysis of the environment of the unusual amino acid residues near the active site indicates that some of the modifications may be required for the enzyme to be catalytically effective. In M. thermoautotrophicum and M. kandleri high temperature adaptation is coupled with increasing intracellular concentrations of lyotropic salts. This was reflected in a higher fraction of glutamate residues at the protein surface of the thermophilic enzymes adapted to high intracellular salt concentrations. PMID- 11023797 TI - The cell biological basis of human implantation. AB - Understanding the cellular basis of implantation and placental development depends on a combination of the limited morphological evidence in the human with data from other primates, separate studies of pre-implantation embryos and endometrium and in vitro models. There is increasing evidence of a dialogue between embryo and endometrium that begins prior to implantation and evolves rapidly through the successive epithelial and stromal/vascular phases. This includes paracrine signals passing to endometrial tissue from the embryo, and vice versa. The production and timing of these signals by endometrium, and its ability to respond to signals from the blastocyst, are dependent on steroidal sensitization. A complex cascade of cell adhesion mechanisms and local tissue remodelling are required for the establishment of a stable haemochorial interface. PMID- 11023798 TI - In vitro models of human blastocyst implantation. AB - This paper reviews different in vitro models used for the study of blastocyst implantation in animals and the human. Furthermore, results from human blastocyst endometrial interactions in vitro, investigated by scanning electron microscopy (SEM), light microscopy (LM) and transmission electron microscopy (TEM), are presented. SEM demonstrates the preference of human blastocysts to adhere to pinopode-presenting areas on endometrial cell cultures. LM and TEM show that the first morphological sign of cell contact, defined as junction formation, is present at the apical-to-lateral border of endometrial epithelial cells, whereas trophoblast attachment to apical endometrial epithelial plasma membranes was not observed. More advanced stages illustrate that the human blastocyst penetrates the epithelial lining by the intrusive penetration mechanism. PMID- 11023799 TI - Endometrial receptivity and the window of implantation. AB - Implantation is a highly co-ordinated event that involves both embryonic and endometrial participation. The endometrium expresses a sophisticated repertoire of proteins during the menstrual cycle many of which help to define a period of receptivity collectively known as the 'window of implantation'. Many of these factors, which are temporally aligned with this window, are now seen as chemical messengers that are recognized by the embryo and facilitate embryonic growth and differentiation. The use of such proteins as biomarkers has also advanced our understanding of the implantation process and may identify women with implantation failure and infertility. While the study of endometrial receptivity is still evolving, the field is growing rapidly and will probably enhance our ability to diagnose and treat couples with infertility, especially in the arena of assisted reproductive technologies (ART). PMID- 11023800 TI - Endocrinology of the peri-implantation period. AB - Current research suggests that the appearance of endometrial integrins and pinopode appearance signal the opening of the receptive phase of the endometrium. These integrins may be activated by the interleukin-1 system (IL-1). IL-1beta, expressed by the blastocyst, induces vascular endothelial growth factor (VEGF) which, in turn, promotes angiogenesis and integrin expression in endometrial cells. The IL-1 system also triggers the expression of gamma interferon (IFN gamma) from T lymphocytes. Decidual natural killer (NK) lymphocytes interact with invading trophoblast to generate leukaemia inhibitory factor (LIF). LIF induces uPA and gelatinase, enzymes which play a crucial role in trophoblastic invasion. Progesterone is a potent inhibitor of LIF, while oestrogen is a potent inducer. Oestrogen in serum reflects follicular IL-1beta level and correlates with the outcome of embryo transfer after in vitro fertilization (IVF). Progesterone induces nitric oxide (NO) synthesis in the decidua, and NO promotes local vasodilatation and uterine quiescenceMeasurement of placental protein 14 (PP14, glycodelin-A) in serum may be of value as a screening test for implantation potential. However, human chorionic gonadotrophin (hCG) remains the most reliable predictor of successful implantation and pregnancy viability. An ovulation + 14 hCG level < 50 IU/l is often predictive of a non-viable outcome, while an ovulation + 21 hCG of < 200 IU/l always indicates a non-viable pregnancy. hCG secretion by invading trophoblast appears to be negatively modulated by endothelin-1 (ET-1) and prostaglandin F(2alpha)(PGF2alpha), while tissue growth factors and collagenases are positive modulators of hCG expression.ProalphaC, an inhibin pro-monomer, may have some value in monitoring corpus luteum function. Inhibin A, activin A and follistatin all rises throughout pregnancy and peak at 36 weeks of gestation. Relaxin is another ovarian hormone that may have a role in predicting implantation. Relaxin induces placental protein 14 (PP14, glycodelin A) expression in a receptive endometrium, and measurement of serum PP14 may be of value as a screening test for implantation potential. PMID- 11023801 TI - Molecular mediators of implantation. AB - Because cytotrophoblastic cells (CTB) from first-trimester placenta form columns of invasive CTB they have been considered as a model for blastocyst implantation. This invasive behaviour is due to the ability of CTB to secret matrix metalloproteinases (MMPs) because tissue inhibitor of MMP (TIMP) inhibits their invasiveness. Although CTB behave like metastatic cells, in vivo they are only transiently invasive (first trimester) and their invasion is normally limited only to the endometrium and to the proximal third of the myometrium. This temporal and spatial regulation of trophoblast invasion is believed to be mediated in an autocrine way by trophoblastic factors and in a paracrine way by uterine factors. Several types of regulators have been investigated: hormones, extracellular matrix (ECM) glycoproteins and cytokines or growth factors. This review is not intended to be an exhaustive catalogue of all the potential regulators but is aimed at describing the mechanism of action of certain factors relevant in trophoblast-endometrial interactions. PMID- 11023802 TI - Paracrine regulators of implantation. AB - Classical and contemporary studies have shown that endocrine regulation exerted by ovarian hormones priming the endometrium is essential for embryo implantation. Increasing evidence indicates that steroid-induced molecules acting as paracrine modulators are necessary for embryo-uterine interactions. That is the case for calcitonin, heparin-binding epidermal growth factor (EGF)-like growth factor, leukaemia inhibitory factor and other molecules. Furthermore, when the blastocyst enters the uterine cavity, it starts the complex signals that will drive embryo adhesion. The paracrinology of this process is based on the local interplay of molecules, such as the secretion of cytokines that may facilitate the acquisition of endometrial receptivity by controlling the expression of adhesion and anti adhesion proteins. Finally, during the embryonic invasive phase, uterine stromal trophoblast dialogue may modulate this self-controlled proteolytic and immunological process to avoid damage to both the embryo and maternal environment. PMID- 11023803 TI - Immunology of implantation. AB - During implantation, the uterine decidua is invaded by extravillous trophoblast (EVT) cells whose function is to destroy the walls of the uterine spiral arteries in order to provide an adequate blood flow to the fetus. These EVT cells express an unusual combination of HLA class I molecules-HLA-C, HLA-E and HLA-G. The decidua is infiltrated by a population of natural killer (NK) cells with a distinctive phenotype of CD56(bright)CD16(-)CD3(-). These cells are particularly numerous in the decidua basalis at the implantation site where they come into close contact with invading EVT cells. These NK cells express a variety of receptors (CD94/NKG2, KIR and ILT) which are known to recognize HLA class I molecules. We believe that interaction between these NK cells and EVT cells provides the controlling influence for implantation. PMID- 11023804 TI - Epidemiology and the medical causes of miscarriage. AB - Human reproduction is extraordinarily wasteful. The reasons for this have taxed all of the contributors to this book. As we move into the 21st century it is sobering to reflect on the fact that we have failed to harness the power of the evolving revolution in molecular medical biology to answer the fundamental question: why is the fate of a fertilized egg so hazardous and so unsuccessful? The following account summarizes our limited knowledge of the epidemiology of miscarriage and then moves on to consider some of the medical causes of miscarriage. The contribution of genetic abnormalities to the problem of pregnancy wastage is discussed elsewhere in this volume. PMID- 11023805 TI - Genetic aspects of miscarriage. AB - Fetal chromosome abnormalities account for about 50% of first-trimester pregnancy losses. Most of these abnormalities are numerical abnormalities (86%) and a low percentage is caused by structural abnormalities (6%) or other genetic mechanisms, including chromosome mosaicism (8%). The recurrence risk of numerical abnormalities is low, so karyotyping of fetal material in case of a miscarriage does not seem worthwhile in daily practice. Half of the structural abnormalities may be inherited from a parent carrying a balanced chromosome translocation or inversion. Parental carriership is found in 4-6% of the couples with recurrent miscarriage. In case of parental carriership of a balanced structural chromosome abnormality, a next pregnancy may result in a child with an unbalanced structural chromosome abnormality. This child can have multiple congenital malformations and/or a mental handicap. Prenatal diagnosis is therefore recommended. Conventional laboratory techniques, such as tissue culturing and karyotyping, or (semi-)direct chromosome technique of chorionic villi, and the recently developed laboratory techniques such as fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH), are described successively. Until now, not enough evidence has been available about the role of other genetic mechanisms, such as single-gene abnormalities, uniparental disomy, genomic imprinting, multifactorial disorders and skewed X chromosome, in the occurrence of miscarriages. PMID- 11023809 TI - Major genes regulating total serum immunoglobulin E levels in families with asthma. AB - Immunoglobulin E (IgE) has a major role in the pathogenesis of allergic disorders and asthma. Previous data from 92 families, each identified through a proband with asthma, showed evidence for two major genes regulating total serum IgE levels. One of these genes mapped to 5q31-33. In the current study, the segregation analysis was extended by the addition of 108 probands and their families, ascertained in the same manner. A mixed recessive model (i.e., major recessive gene and residual genetic effect) was the best-fitting and most parsimonious one-locus model of the segregation analysis. A mixed two-major-gene model (i.e., two major genes and residual genetic effect) fit the data significantly better than did the mixed recessive one-major-gene model. The second gene modified the effect of the first recessive gene. Individuals with the genotype aaBB (homozygous high-risk allele at the first gene and homozygous low risk allele at the second locus) had normal IgE levels (mean 23 IU/ml), and only individuals with genotypes aaBb and aabb had high IgE levels (mean 282 IU/ml). A genomewide screening was performed using variance-component analysis. Significant evidence for linkage was found for a novel locus at 7q, with a multipoint LOD score of 3. 36 (P=.00004). A LOD score of 3.65 (P=.00002) was obtained after genotyping additional markers in this region. Evidence for linkage was also found for two previously reported regions, 5q and 12q, with LOD scores of 2.73 (P=.0002) and 2.46 (P=.0004), respectively. These results suggest that several major genes, plus residual genetic effects, regulate total serum IgE levels. PMID- 11023810 TI - Human nonsyndromic hereditary deafness DFNA17 is due to a mutation in nonmuscle myosin MYH9. AB - The authors had previously mapped a new locus-DFNA17, for nonsyndromic hereditary hearing impairment-to chromosome 22q12.2-q13. 3. DFNA17 spans a 17- to 23-cM region, and MYH9, a nonmuscle-myosin heavy-chain gene, is located within the linked region. Because of the importance of myosins in hearing, MYH9 was tested as a candidate gene for DFNA17. Expression of MYH9 in the rat cochlea was confirmed using reverse transcriptase-PCR and immunohistochemistry. MYH9 was immunolocalized in the organ of Corti, the subcentral region of the spiral ligament, and the Reissner membrane. Sequence analysis of MYH9 in a family with DFNA17 identified, at nucleotide 2114, a G-->A transposition that cosegregated with the inherited autosomal dominant hearing impairment. This missense mutation changes codon 705 from an invariant arginine (R) to histidine (H), R705H, within a highly conserved SH1 linker region. Previous studies have shown that modification of amino acid residues within the SH1 helix causes dysfunction of the ATPase activity of the motor domain in myosin II. Both the precise role of MYH9 in the cochlea and the mechanism by which the R705H mutation leads to the DFNA17 phenotype (progressive hearing impairment and cochleosaccular degeneration) remain to be elucidated. PMID- 11023811 TI - Messages through bottlenecks: on the combined use of slow and fast evolving polymorphic markers on the human Y chromosome. PMID- 11023812 TI - Asymmetries in the maternal and paternal genetic histories of Colombian populations. PMID- 11023814 TI - AMP decreases the efficiency of skeletal-muscle mitochondria. AB - Mitochondrial proton leak in rat muscle is responsible for approx. 15% of the standard metabolic rate, so its modulation could be important in regulating metabolic efficiency. We report in the present paper that physiological concentrations of AMP (K(0.5)=80 microM) increase the resting respiration rate and double the proton conductance of rat skeletal-muscle mitochondria. This effect is specific for AMP. AMP also doubles proton conductance in skeletal muscle mitochondria from an ectotherm (the frog Rana temporaria), suggesting that AMP activation is not primarily for thermogenesis. AMP activation in rat muscle mitochondria is unchanged when uncoupling protein-3 is doubled by starvation, indicating that this protein is not involved in the AMP effect. AMP activation is, however, abolished by inhibitors and substrates of the adenine nucleotide translocase (ANT), suggesting that this carrier (possibly the ANT1 isoform) mediates AMP activation. AMP activation of ANT could be important for physiological regulation of metabolic rate. PMID- 11023815 TI - Impaired reductive regeneration of ascorbic acid in the Goto-Kakizaki diabetic rat. AB - Ascorbic acid (AA) is a naturally occurring major antioxidant that is essential for the scavenging of toxic free radicals in both plasma and tissues. AA levels in plasma and tissues have been reported to be significantly lower than normal in diabetic animals and humans, and might contribute to the complications found at the late stages of diabetes. In this study, plasma and hepatic AA levels and AA regeneration were studied in the Goto-Kakizaki diabetic rat (GK rat) to elucidate the mechanism of decreasing plasma and hepatic AA levels in diabetes. AA concentrations in the plasma and liver were significantly lower in GK than in control rats. AA levels in primary cultured hepatocytes derived from GK rats were lower than those derived from control Wistar rats with or without dehydroascorbic acid (DHA) in the medium. Among various enzyme activities that reduce DHA to AA, the NADPH-dependent regeneration of AA in the liver was significantly suppressed in GK rats. Northern blot analysis revealed that only the expression of 3-alpha hydroxysteroid dehydrogenase (AKR) was significantly suppressed in these rats. These results suggest that decreased AA-regenerating activity, probably through decreased expression of AKR, contributes to the decreased AA levels and increased oxidative stress in GK rats. PMID- 11023813 TI - Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. AB - The Ras/Raf/MEK (mitogen-activated protein kinase/ERK kinase)/ERK (extracellular signal-regulated kinase) pathway is at the heart of signalling networks that govern proliferation, differentiation and cell survival. Although the basic regulatory steps have been elucidated, many features of this pathway are only beginning to emerge. This review focuses on the role of protein-protein interactions in the regulation of this pathway, and how they contribute to co ordinate activation steps, subcellular redistribution, substrate phosphorylation and cross-talk with other signalling pathways. PMID- 11023816 TI - Anti-(herpes simplex virus) activity of 4'-thio-2'-deoxyuridines: a biochemical investigation for viral and cellular target enzymes. AB - The antiviral activity of several nucleoside analogues is often limited by their rapid degradation by pyrimidine nucleoside phosphorylases. In an attempt to avoid this degradation, several modified nucleosides have been synthesized. A series of 4'-thio-2'-deoxyuridines exhibits an anti-[herpes simplex virus (HSV)] activity significantly higher (20-600 times) than that shown by the corresponding 4'-oxy counterpart. We investigated the mode of action of these compounds and we found that: (i) several 4'-thio-2'-deoxyuridines are phosphorylated to the mono- and di phosphates by HSV-1 thymidine kinase (TK) more efficiently than their corresponding 4'-oxy counterpart; (ii) both are inhibitors of cellular thymidylate synthase; (iii) 4'-thio-2'-deoxyuridines are resistant to phosphorolysis by human thymidine phosphorylase; (iv) both 4'-oxy- and 4'-thio-2' deoxyuridines are phosphorylated to deoxyribonucleotide triphosphate in HSV-1 infected cells and are incorporated into viral DNA; (v) 4'-thio-2'-deoxyuridines are better inhibitors than their 4'-oxy counterparts of [(3)H]thymidine incorporation in HSV-1-infected cells; (vi) 4'-thio-2'-deoxyuridines are not recognized by HSV-1 and human uracil-DNA glycosylases. Our data suggest that 4' thio-2'-deoxyuridines, resistant to pyrimidine phosphorylase, can be preferentially or selectively phosphorylated by viral TK in HSV-infected cells, where they are further converted into triphosphate by cellular nucleotide kinases. Once incorporated into viral DNA, they are better inhibitors of viral DNA synthesis than their corresponding 4'-oxy counterpart, either because they are not recognized, and thus not removed, by viral uracil-DNA glycosylase, or because they preferentially interfere with viral DNA polymerase. PMID- 11023817 TI - Ischaemia induces changes in the association of the binding protein 4E-BP1 and eukaryotic initiation factor (eIF) 4G to eIF4E in differentiated PC12 cells. AB - Ischaemia was obtained in vitro by subjecting nerve-growth-factor-differentiated PC12 cells to glucose deprivation plus anoxia. During ischaemia the rate of protein synthesis was significantly inhibited, and eIF4E-binding protein (4E-BP1) and eukaryotic initiation factor 4E (eIF4E) were significantly dephosphorylated in parallel. In addition, ischaemia induced an enhancement of the association of 4E-BP1 to eIF4E, which in turn decreased eIF4F formation, whereas no degradation of initiation factor 4G was observed. The treatment of PC12 cells with the specific p38 mitogen-activated protein kinase inhibitor SB203580 induced eIF4E dephosphorylation but did not cause any effect on protein synthesis rate. Rapamycin, the inhibitor of mammalian target of rapamycin ('mTOR'), but not PD98059, the inhibitor of extracellular signal-regulated protein kinases ('ERK1/2'), induced similar effects on 4E-BP1 phosphorylation to ischaemia; nevertheless, 4E-BP1-eIF4E complex levels were higher in ischaemia than in rapamycin-treated cells. In addition, both protein synthesis rate and eIF4F formation were lower in ischaemic cells than in rapamycin-treated cells. PMID- 11023818 TI - 'pH-jump' crystallographic analyses of gamma-lactam-porcine pancreatic elastase complexes. AB - beta-Lactams inhibit a range of enzymes via acylation of nucleophilic serine residues. Certain gamma-lactam analogues of monocyclic beta-lactams have also been shown to be reversible inhibitors of porcine pancreatic elastase (PPE), forming acyl-enzyme complexes that are stable with respect to hydrolysis. Crystallographic analysis at pH 5 of an acyl-enzyme complex formed with PPE and one of these inhibitors revealed the ester carbonyl located in the oxyanion hole in a similar conformation to that observed in the structure of a complex formed between a heptapeptide (beta-casomorphin-7) and PPE. Only weak electron density was observed for the His-57 side chain in its 'native' conformation. Instead, the His-57 side chain predominantly adopted a conformation rotated approx. 90 degrees from its normal position. PPE-gamma-lactam crystals were subjected to 'pH-jumps' by placing the crystals in a buffer of increased pH prior to freezing for data collection. The results indicate that the conformation of the gamma-lactam derived acyl-enzyme species in the PPE active site is dependent on pH, a result having implications for the analysis of other serine protease-inhibitor structures at non-catalytic pH values. The results help to define the stereoelectronic relationship between the ester of the acyl-enzyme complex, the side chain of His-57 and the incoming nucleophile during the reversible (de)acylation steps, implying it is closely analogous to the hydrolytic deacylation step during catalytic peptide hydrolysis. PMID- 11023819 TI - Evaluation of the role of two conserved active-site residues in beta class glutathione S-transferases. AB - Glutathione S-transferases (GSTs) normally use hydroxy-group-containing residues in the N-terminal domain of the enzyme for stabilizing the activated form of the co-substrate, glutathione. However, previous mutagenesis studies have shown that this is not true for Beta class GSTs and thus the origin of the stabilization remains a mystery. The recently determined crystal structure of Proteus mirabilis GST B1-1 (PmGST B1-1) suggested that the stabilizing role might be fulfilled in Beta class GSTs by one or more residues in the C-terminal domain of the enzyme. To test this hypothesis we mutated His(106) and Lys(107) of PmGST B1-1 to investigate their possible role in the enzyme's catalytic activity. His(106) was mutated to Ala, Asn and Phe, and Lys(107) to Ala and Arg. The effects of the replacement on the activity, thermal stability and antibiotic-binding capacity of the enzyme were examined. The results are consistent with the involvement of His(106) and Lys(107) in interacting with glutathione at the active site but these residues do not contribute significantly to catalysis, folding or antibiotic binding. PMID- 11023820 TI - Receptor-activity-modifying protein 1 forms heterodimers with two G-protein coupled receptors to define ligand recognition. AB - Receptor-activity-modifying proteins (RAMPs) with single transmembrane domains define the function of two G-protein-coupled receptors of the B family. Cell surface complexes of human RAMP1 (hRAMP1) and human calcitonin (CT) receptor isotype 2 (hCTR2) or rat CT-receptor-like receptor (rCRLR) have now been identified through protein cross-linking, co-immunoprecipitation and confocal microscopy. They are two distinct CT-gene-related peptide (CGRP) receptors coupled to cAMP production and pharmacologically distinguished by the CT and CGRP antagonists salmon CT(8-32) and human or rat CGRP(8-37). Thus direct molecular interactions of hRAMP1 with hCTR2 or rCRLR are required for CGRP recognition. hCTR2, moreover, adopts non-traditional functions through its association with hRAMP1. PMID- 11023821 TI - Identification and characterization of a silencer regulatory element in the 3' flanking region of the murine CD46 gene. AB - The murine membrane cofactor protein (CD46) gene is expressed exclusively in testis, in contrast to human CD46, which is expressed ubiquitously. To elucidate the mechanism of differential CD46 gene expression among species, we cloned entire murine CD46 genomic DNA and possible regulatory regions were placed in the flanking region of the luciferase reporter gene. The reporter gene assay revealed a silencing activity not in the promoter, but in the 3'-flanking region of the gene and the silencer-like element was identified within a 0.2-kb region between 0.6 and 0.8 kb downstream of the stop codon. This silencer-like element was highly similar to that of the pig MHC class-I gene. The introduction of a mutation into this putative silencer element of murine CD46 resulted in an abrogation of the silencing effect. Electrophoretic mobility-shift assay indicated the presence of the binding molecule(s) for this silencer sequence in murine cell lines and tissues. A size difference of the protein-silencer-element complex was observed depending upon the solubilizers used for preparation of the nuclear extracts. A mutated silencer sequence failed to interact with the binding molecules. The level of the binding factor was lower in the testicular germ cells compared with other organs. Thus the silencer element and its binding factor may play a role in transcriptional regulation of murine CD46 gene expression. These results imply that the effects of the CD46 silencer element encompass the innate immune and reproductive systems, and in mice may determine the testicular germ cell-dominant expression of CD46. PMID- 11023822 TI - The tissue-specific regulation of the carboxyl ester lipase gene in exocrine pancreas differs significantly between mouse and human. AB - The carboxyl ester lipase (CEL) gene is highly expressed in exocrine pancreas and expression of the human CEL gene is mediated by a strong tissue-specific enhancer, which is absolutely necessary for high-level expression. The mouse promoter, on the other hand, does not contain a corresponding enhancer element, but instead is totally dependent on another pancreas-specific element. This element is identified as a pancreatic transcription factor 1 (PTF 1)-binding site. The human CEL promoter also contains a putative PTF 1 element located at a position corresponding to the essential PTF 1 site in the mouse promoter. However, nucleotide changes in the human promoter 5' flanking this PTF 1 site have created an overlapping CCAAT/enhancer-binding protein (C/EBP)-like binding motif, interfering with the binding of PTF 1. Hence, our findings provide an example of genetic divergence between species not accompanied by difference in function. PMID- 11023823 TI - A mammalian cytochrome fused to a chloroplast transit peptide is a functional haemoprotein and is imported into isolated chloroplasts. AB - The small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a major chloroplast stromal protein that is cytosolically synthesized as a precursor with an N-terminal extension, known as the transit sequence or transit peptide (Tp). The Tp is essential for the post-translational uptake of the precursor by the chloroplast. The Tp is thought to influence the conformation of the precursor protein and to facilitate polypeptide translocation across the chloroplast envelope barrier via a Tp-selective translocon. To address these issues we have devised a novel strategy to generate substrate amounts of a chloroplast targeting sequence as a fusion with the chromogenic globular domain of cytochrome b(5) (Cyt). The chimaeric protein is an ideal probe for investigating the conformation of a preprotein and events surrounding protein import into isolated chloroplasts. The Cyt of liver endoplasmic reticulum was fused at its N-terminus with the Tp of the small subunit of Rubisco of Pisum sativum (pea). To enhance its production by clearance from the cytoplasm of Escherichia coli, the chimaera was engineered by further N-terminal linkage of a prokaryotic secretory signal. Expression of this tripartite fusion resulted in mg quantities of the signal sequence-processed Tp-Cyt protein, which was eventually targeted to the membranes. The chromogenic nature of the chimaera and its localization to the bacterial membrane facilitated the biochemical isolation of the precursor in a soluble and functional form. The purified preprotein displayed spectral and enzymic properties that were indistinguishable from the native parental Cyt, implying an absence of observable influence of the Tp on the conformation of the haemoprotein. The chimaeric precursor was imported into the stroma of the isolated chloroplasts in a dose-dependent manner. Import was also strongly dependent upon exogenously supplied ATP. The stromally imported chimaeric precursor protein was processed to a size characteristic of Cyt. PMID- 11023824 TI - Characterization of the human liver fructose-1,6-bisphosphatase gene promoter. AB - Fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11), an important gluconeogenic enzyme, catalyses the hydrolysis of fructose 1, 6-bisphosphate to fructose 6 phosphate and P(i). Enzyme activity is mainly regulated by the allosteric inhibitors fructose 2, 6-bisphosphate and AMP. Although some observations about hormonal regulation of the enzyme have been published, the FBPase promoter has not been studied in detail. Here we report an in vitro characterization of the FBPase promoter with respect to the elements that are required for basal promoter activity. Transient transfection of H4IIE rat hepatoma cells, combined with site directed mutagenesis, demonstrated that an enhancer box, three GC-boxes and a nuclear factor kappaB (NF-kappaB)-binding element are important for hepatic FBPase promoter activity. These elements are found in the region located between 405 to +25 bp relative to the transcription start site. Electrophoretic-mobility shift assays and supershift analysis confirmed that upstream stimulatory factor 1 (USF1)/USF2, specificity protein 1 (Sp1)/Sp3 and NF-kappaB respectively bind to these sites. The present study provides the basis for a more comprehensive screening for mutations in FBPase-deficient patients and for further studies of the transcriptional regulation of this gene. PMID- 11023825 TI - DNA repair protein O6-alkylguanine-DNA alkyltransferase is phosphorylated by two distinct and novel protein kinases in human brain tumour cells. AB - We showed recently that human O(6)-alkylguanine-DNA alkyltransferase (AGT), an important target for improving cancer chemotherapy, is a phosphoprotein and that phosphorylation inhibits its activity [Srivenugopal, Mullapudi, Shou, Hazra and Ali-Osman (2000) Cancer Res. 60, 282-287]. In the present study we characterized the cellular kinases that phosphorylate AGT in the human medulloblastoma cell line HBT228. Crude cell extracts used Mg(2+) more efficiently than Mn(2+) for phosphorylating human recombinant AGT (rAGT) protein. Both [gamma-(32)P]ATP and [gamma-(32)P]GTP served as phosphate donors, with the former being twice as efficient. Specific components known to activate protein kinase A, protein kinase C and calmodulin-dependent kinases did not stimulate the phosphorylation of rAGT. Phosphoaminoacid analysis after reaction in vitro with ATP or GTP showed that AGT was modified at the same amino acids (serine, threonine and tyrosine) as in intact HBT228 cells. Although some of these properties pointed to casein kinase II as a candidate enzyme, known inhibitors and activators of casein kinase II did not affect rAGT phosphorylation. Fractionation of the cell extracts on poly(Glu/Tyr)-Sepharose resulted in the adsorption of an AGT kinase that modified the tyrosine residues and the exclusion of a fraction that phosphorylated AGT on serine and threonine residues. In-gel kinase assays after SDS/PAGE and non denaturing PAGE revealed the presence of two AGT kinases of 75 and 130 kDa in HBT228 cells. The partly purified tyrosine kinase, identified as the 130 kDa enzyme by the same assays, was strongly inhibited by tyrphostin 25 but not by genestein. The tyrosine kinase used ATP or GTP to phosphorylate the AGT protein; this reaction inhibited the DNA repair activity of AGT. Evidence that the kinases might physically associate with AGT in cells was also provided. These results demonstrate that two novel cellular protein kinases, a tyrosine kinase and a serine/threonine kinase, both capable of using GTP as a donor, phosphorylate the AGT protein and affect its function. The new kinases might serve as potential targets for strengthening the biochemical modulation of AGT in human tumours. PMID- 11023826 TI - Molecular characterization of human tensin. AB - Tensin is a focal-adhesion molecule that binds to actin filaments and interacts with phosphotyrosine-containing proteins. To analyse tensin's function in mammals, we have cloned tensin cDNAs from human and cow. The isolated approx. 7.7 kb human cDNA contains an open reading frame encoding 1735 amino acid residues. The amino acid sequence of human tensin shares 60% identity with chicken tensin, and contains all the structural features described previously in chicken tensin. This includes the actin-binding domains, the Src homology domain 2, and the region similar to a tumour suppressor, PTEN. Two major differences between human and chicken tensin are (i) the lack of the first 54 residues present in chicken tensin, and (ii) the addition of 34- and 38-residue inserts in human and bovine tensin. In addition, our interspecies sequencing data have uncovered the presence of a glutamine/CAG repeat that appears to have expanded in the course of evolution. Northern-blot analysis reveals a 10-kb message in most of the human tissues examined. An additional 9-kb message is detected in heart and skeletal muscles. The molecular mass predicted from the human cDNA is 185 kDa, although both endogenous and recombinant human tensin migrate as 220-kDa proteins on SDS/PAGE. The discrepancy is due to the unusually low electrophoretic mobility of the central region of the tensin polypeptide (residues 306-981). A survey of human prostate and breast cancer cell lines by Western-blot analysis shows a lack of tensin expression in most cancer cell lines, whereas these lines express considerable amounts of focal-adhesion molecules such as talin and focal-adhesion kinase. Finally, tensin is rapidly cleaved by a focal-adhesion protease, calpain II. Incubation of cells with a calpain inhibitor, MDL, prevented tensin cleavage and induced morphological change in these cells, suggesting that cleavage of tensin and other focal-adhesion constituents by calpain disrupts maintenance of normal cell shape. PMID- 11023827 TI - Regulation of calpain and calpastatin in differentiating myoblasts: mRNA levels, protein synthesis and stability. AB - Calpain (Ca(2+)-dependent intracellular protease)-induced proteolysis has been considered to play a role in myoblast fusion to myotubes. We found previously that calpastatin (the endogenous inhibitor of calpain) diminishes transiently during myoblast differentiation. To gain information about the regulation of calpain and calpastatin in differentiating myoblasts, we evaluated the stability and synthesis of calpain and calpastatin, and measured their mRNA levels in L8 myoblasts. We show here that mu-calpain and m-calpain are stable, long-lived proteins in both dividing and differentiating L8 myoblasts. Calpain is synthesized in differentiating myoblasts, and calpain mRNA levels do not change during differentiation. In contrast, calpastatin (though also a long-lived protein in myoblasts), is less stable in differentiating myoblasts than in the dividing cells, and its synthesis is inhibited upon initiation of differentiation. Inhibition of calpastatin synthesis is followed by a diminution in calpastatin mRNA levels. A similar calpastatin mRNA diminution is observed upon drug-induced inhibition of protein translation. On the other hand, transforming growth factor beta (which inhibits differentiation) allows calpastatin synthesis and prevents the diminution in calpastatin mRNA. The overall results suggest that at the onset of myoblast differentiation, calpastatin is regulated mainly at the level of translation and that an inhibition of calpastatin synthesis leads to the decrease in its mRNA stability. The existing calpastatin then diminishes, resulting in decreased calpastatin activity in the fusing myoblasts, allowing calpain activation and protein degradation required for fusion. PMID- 11023828 TI - Macromolecular organization of saliva: identification of 'insoluble' MUC5B assemblies and non-mucin proteins in the gel phase. AB - Stimulated human submandibular/sublingual (HSMSL) and whole saliva were separated into sol and gel phases and mucins were isolated by density-gradient centrifugation in CsCl/4M guanidinium chloride. MUC5B and MUC7 were identified using anti-peptide antisera raised against sequences within the MUC5B and MUC7 apoproteins respectively. MUC7 was found mainly in the sol phase of both HSMSL and whole saliva, but some MUC7 was consistently present in the gel phase, suggesting that this mucin may interact with the salivary gel matrix. In HSMSL saliva, MUC5B was found in the gel phase; however, most of the material was 'insoluble' in guanidinium chloride and was only brought into solution by reduction. In whole saliva, the MUC5B mucin was present both in the sol and gel phases although some material was again 'insoluble'. Rate-zonal centrifugation of whole saliva showed that MUC5B mucins in the sol phase were smaller than those in the gel phase, suggesting differences in oligomerization and/or degradation. Antibodies against IgA, secretory component, lysozyme and lactoferrin were used to study the distribution of non-gel-forming proteins in the different phases of saliva. The majority of these proteins was found in the sol phase of both HSMSL and whole saliva. However, a significant fraction was present in the gel phase of whole saliva, suggesting a post-secretory interaction with the salivary gel matrix. A monoclonal antibody against a parotid salivary agglutinin was used to show that this protein is present mainly in the gel phase of both whole saliva and parotid secretion. PMID- 11023829 TI - Tyrosine kinases activate store-mediated Ca2+ entry in human platelets through the reorganization of the actin cytoskeleton. AB - We have recently reported that store-mediated Ca(2+) entry in platelets is likely to be mediated by a reversible trafficking and coupling of the endoplasmic reticulum with the plasma membrane, a model termed 'secretion-like coupling'. In this model the actin cytoskeleton plays a key regulatory role. Since tyrosine kinases have been shown to be important for Ca(2+) entry in platelets and other cells, we have now investigated the possible involvement of tyrosine kinases in the secretion-like-coupling model. Treatment of platelets with thrombin or thapsigargin induced actin polymerization by a calcium-independent pathway. Methyl 2,5-dihydroxycinnamate, a tyrosine kinase inhibitor, prevented thrombin- or thapsigargin-induced actin polymerization. The effects of tyrosine kinases in store-mediated Ca(2+) entry were found to be entirely dependent on the actin cytoskeleton. PP1, an inhibitor of the Src family of proteins, partially inhibited store-mediated Ca(2+) entry. In addition, depletion of intracellular Ca(2+) stores stimulated cytoskeletal association of the cytoplasmic tyrosine kinase pp60(src), a process that was sensitive to treatment with cytochalasin D and PP1, but not to inhibition of Ras proteins using prenylcysteine analogues. Finally, combined inhibition of both Ras proteins and tyrosine kinases resulted in complete inhibition of Ca(2+) entry, suggesting that these two families of proteins have independent effects in the activation of store-mediated Ca(2+) entry in human platelets. PMID- 11023830 TI - Effect of spermine synthase deficiency on polyamine biosynthesis and content in mice and embryonic fibroblasts, and the sensitivity of fibroblasts to 1,3-bis-(2 chloroethyl)-N-nitrosourea. AB - Mutant Gy male mice, which have previously been described as having disruption of the phosphate-regulating Phex gene and a spermine synthase gene [Meyer, Henley, Meyer, Morgan, McDonald, Mills and Price (1998) Genomics, 48, 289-295; Lorenz, Francis, Gempel, Boddrich, Josten, Schmahl and Schmidt (1998) Hum. Mol. Genet. 7, 541-547], as well as mutant Hyp male mice, which have disruption of the Phex gene only, were examined along with their respective normal male littermates. Biochemical analyses of extracts of brains, hearts and livers of 5-week-old mice showed that Gy males lacked any significant spermine synthase activity as well as spermine content. Organs of Gy males had a higher spermidine content. This was caused not only by the lack of conversion of spermidine into spermine, but also because of compensatory increases in the activities of other polyamine biosynthetic enzymes. Gy males were half the body weight of their normal male littermates at weaning age. Hyp males, however, were no different in size when compared with their controls. High mortality of Gy males occurs by weaning age and this mortality was shown to be largely post-natal. Embryonic fibroblasts were isolated from Gy males and their normal male littermates and were similarly shown to lack any significant spermine synthase activity as well as spermine content. The lack of spermine, however, had no significant effect on the growth of immortalized fibroblasts or of primary fibroblast cultures. Similarly, there was no difference in the time of senescence of primary fibroblast cultures from Gy males compared with cultures derived from normal male littermates. However, the lack of spermine did increase the sensitivity of immortalized fibroblasts to killing by the chloroethylating agent 1, 3-bis(2-chloroethyl)-N-nitrosourea. Therefore both the Gy male mice and derived embryonic fibroblasts provide valuable models to study the importance of spermine and spermine synthase, without the use of inhibitors which may have additional side effects. PMID- 11023831 TI - Mechanism of binding of surfactant protein D to influenza A viruses: importance of binding to haemagglutinin to antiviral activity. AB - Collectins are important in the initial containment of a variety of pathogens, including influenza A virus (IAV). We provide the first systematic evaluation of the oligosaccharide-binding sites for pulmonary surfactant protein D (SP-D) on specific IAV coat glycoproteins and define the relationship between this binding and antiviral activity. With the use of several techniques, SP-D was found to bind via its carbohydrate-recognition domain (CRD) to mannosylated, N-linked carbohydrates on the HA(1) domain of the haemagglutinin (HA) and on the neuraminidase of IAV. Using a set of IAV strains that differed in the level and site of glycosylation, and a panel of recombinant collectins, we found that binding of SP-D to the globular domain of the HA was critical in mediating the inhibition of viral haemagglutination activity and infectivity. We also demonstrated that the pattern of binding of a collectin to IAV glycoproteins can be modified by altering the monosaccharide-binding affinity of its CRD or by linking the CRD to a different N-terminal/collagen domain. These studies clarify the mechanisms of viral neutralization by collectins and might be useful in engineering collectins for enhanced antiviral activity. PMID- 11023833 TI - Cloning and characterization of full-length mouse thymidine kinase 2: the N terminal sequence directs import of the precursor protein into mitochondria. AB - The subcellular localization of mitochondrial thymidine kinase (TK2) has been questioned, since no mitochondrial targeting sequences have been found in cloned human TK2 cDNAs. Here we report the cloning of mouse TK2 cDNA from a mouse full length enriched cDNA library. The mouse TK2 cDNA codes for a protein of 270 amino acids, with a 40-amino-acid presumed N-terminal mitochondrial targeting signal. In vitro translation and translocation experiments with purified rat mitochondria confirmed that the N-terminal sequence directed import of the precursor TK2 into the mitochondrial matrix. A single 2.4 kb mRNA transcript was detected in most tissues examined, except in liver, where an additional shorter (1.0 kb) transcript was also observed. There was no correlation between the tissue distribution of TK2 activity and the expression of TK2 mRNA. Full-length mouse TK2 protein and two N-terminally truncated forms, one of which corresponds to the mitochondrial form of TK2 and a shorter form corresponding to the previously characterized recombinant human TK2, were expressed in Escherichia coli and affinity purified. All three forms of TK2 phosphorylated thymidine, deoxycytidine and 2'-deoxyuridine, but with different kinetic efficiencies. A number of cytostatic pyrimidine nucleoside analogues were also tested and shown to be good substrates for the various forms of TK2. The active form of full-length mouse TK2 was a dimer, as judged by Superdex 200 chromatography. These results enhance our understanding of the structure and function of TK2, and may help to explain the mitochondrial disorder, mitochondrial neurogastrointestinal encephalomyopathy. PMID- 11023832 TI - Phosphatidylcholine-specific phospholipase C and phospholipase D are respectively implicated in mitogen-activated protein kinase and nuclear factor kappaB activation in tumour-necrosis-factor-alpha-treated immature acute-myeloid leukaemia cells. AB - Tumour necrosis factor-alpha (TNFalpha) has been reported to induce potent growth inhibition of committed myeloid progenitor cells, whereas it is a potential growth stimulator of human CD34(+)CD38(-) multipotent haematopoietic cells. The present study was aimed at evaluating the respective role of two phospholipases, phosphatidylcholine-specific phospholipase C (PC-PLC) and phospholipase D (PLD) in the response of the CD34(+) CD38(-) KG1a cells to TNFalpha. In these cells TNFalpha triggered phosphoinositide 3-kinase (PI3K)-dependent PC hydrolysis within 4-8 min with concomitant production of both diacylglycerol (DAG) and phosphocholine (P-chol). DAG and P-chol production was accompanied by extracellular-signal-related protein kinase-1 ('ERK-1') activation and DNA synthesis stimulation. PC-PLC stimulation was followed by PI3K-independent PLD activation with concomitant phosphatidic acid (PA) production followed by PA derived DAG accumulation and sustained nuclear factor kappaB (NF-kappaB) activation. PLD/NF-kappaB signalling activation played no role in the TNFalpha proliferative effect and conferred no consistent protection of KG1a cells towards antileukaemic agents. Altogether these results suggest that, in KG1a cells, TNFalpha can stimulate in parallel PC-PLC and PLD, whose lipid products activate in turn mitogen-activated protein kinase (MAP kinase) and NF-kappaB signalling respectively. Finally, our study suggests that PC-PLC, but not PLD, plays a role in the TNFalpha proliferative effect in immature myeloid cells. PMID- 11023834 TI - The Fe(II) permease Fet4p functions as a low affinity copper transporter and supports normal copper trafficking in Saccharomyces cerevisiae. AB - The plasma-membrane of Saccharomyces cerevisiae contains high affinity permeases for Cu(I) and Fe(II). A low affinity Fe(II) permease has also been identified, designated Fet4p. A corresponding low affinity copper permease has not been characterized, although yeast cells that lack high affinity copper uptake do accumulate this metal ion. We demonstrate in the present study that Fet4p can function as a low affinity copper permease. Copper is a non-competitive inhibitor of (55)Fe uptake through Fet4p (K(i)=22 microM). Fet4p-dependent (67)Cu uptake was kinetically characterized, with K(m) and V(max) values of 35 microM and 8 pmol of copper/min per 10(6) cells respectively. A fet4-containing strain exhibited no saturable, low affinity copper uptake indicating that this uptake was attributable to Fet4p. Mutant forms of Fet4p that exhibited decreased efficiency in (55/59)Fe uptake were similarly compromised in (67)Cu uptake, indicating that similar amino acid residues in Fet4p contribute to both uptake processes. The copper taken into the cell by Fet4p was metabolized similarly to the copper taken into the cell by the high affinity permease, Ctr1p. This was shown by the Fet4p-dependence of copper activation of Fet3p, the copper oxidase that supports high affinity iron uptake in yeast. Also, copper-transported by Fet4p down-regulated the copper sensitive transcription factor, Mac1p. Whether supplied by Ctr1p or by Fet4p, an intracellular copper concentration of approx. 10 microM caused a 50% reduction in the transcriptional activity of Mac1p. The data suggest that the initial trafficking of newly arrived copper in the yeast cell is independent of the copper uptake pathway involved, and that this copper may be targeted first to a presumably small 'holding' pool prior to its partitioning within the cell. PMID- 11023835 TI - Nitric oxide inhibits isoproterenol-stimulated adipocyte lipolysis through oxidative inactivation of the beta-agonist. AB - Nitric oxide has been implicated in the inhibition of catecholamine-stimulated lipolysis in adipose tissue by as yet unknown mechanisms. In the present study, it is shown that the nitric oxide donor, 2,2-diethyl-1-nitroso-oxyhydrazine, antagonized isoproterenol (isoprenaline)-induced lipolysis in rat adipocytes, freshly isolated from white adipose tissue, by decreasing the potency of the beta agonist without affecting its efficacy. These data suggest that nitric oxide did not act downstream of the beta-adrenoceptor but reduced the effective concentration of isoproterenol. In support of the latter hypothesis, we found that pre-treatment of isoproterenol with nitric oxide abolished the lipolytic activity of the catecholamine. Spectroscopic data and HPLC analysis confirmed that the nitric oxide-mediated inactivation of isoproterenol was in fact because of the modification of the catecholamine through a sequence of oxidation reactions, which apparently involved the generation of an aminochrome. Similarly, aminochrome was found to be the primary product of isoproterenol oxidation by 3 morpholinosydnonimine and peroxynitrite. Finally, it was shown that nitric oxide released from cytokine-stimulated adipocytes attenuated the lipolytic effect of isoproterenol by inactivating the catecholamine. In contrast with very recent findings, which suggest that nitric oxide impairs the beta-adrenergic action of isoproterenol through intracellular mechanisms and not through a chemical reaction between NO and the catecholamine, we showed that nitric oxide was able to attenuate the pharmacological activity of isoproterenol in vitro as well as in a nitric oxide-generating cellular system through oxidation of the beta-agonist. These findings should be taken into account in both the design and interpretation of studies used to investigate the role of nitric oxide as a modulator of isoproterenol-stimulated signal transduction pathways. PMID- 11023836 TI - Inhibition by etomoxir of rat liver carnitine octanoyltransferase is produced through the co-ordinate interaction with two histidine residues. AB - Rat peroxisomal carnitine octanoyltransferase (COT), which facilitates the transport of medium-chain fatty acids through the peroxisomal membrane, is irreversibly inhibited by the hypoglycaemia-inducing drug etomoxir. To identify the molecular basis of this inhibition, cDNAs encoding full-length wild-type COT, two different variant point mutants and one variant double mutant from rat peroxisomal COT were expressed in Saccharomyces cerevisiae, an organism devoid of endogenous COT activity. The recombinant mutated enzymes showed activity towards both carnitine and decanoyl-CoA in the same range as the wild type. Whereas the wild-type version expressed in yeast was inhibited by etomoxir in an identical manner to COT from rat liver peroxisomes, the activity of the enzyme containing the double mutation H131A/H340A was completely insensitive to etomoxir. Individual point mutations H131A and H340A also drastically reduced sensitivity to etomoxir. Taken together, these results indicate that the two histidine residues, H131 and H340, are the sites responsible for inhibition by etomoxir and that the full inhibitory properties of the drug will be shown only if both histidines are intact at the same time. Our data demonstrate that both etomoxir and malonyl-CoA inhibit COT by interacting with the same sites. PMID- 11023837 TI - alpha2-macroglobulin modulates the immunoregulatory function of the lipocalin placental protein 14. AB - Human placental protein 14 (PP14; also known as glycodelin and progesterone associated endometrial protein) is an immunosuppressive protein of the lipocalin structural superfamily. Mechanisms regulating serum PP14's immunosuppressive activity remain to be elucidated. In the present study, an interaction between PP14 and a major serum protein carrier, alpha(2)-macroglobulin (alpha(2)M), was documented for the first time. Using native gel electrophoresis, we showed that PP14, as well as its alternative splice variant PP14.2, binds to both alpha(2)M and methylamine-activated (MA)-alpha(2)M. Cross-competition studies demonstrated that the variants compete for binding to alpha(2)M. PP14 bound to alpha(2)M and MA-alpha(2)M with K(d) values of 167+/-70 and 221+/-56 nM (means+/-S.D.) respectively, as determined by surface plasmon resonance. Significantly, the addition of alpha(2)M or MA-alpha(2)M to a T-cell proliferation assay strongly potentiated the inhibitory capacity of PP14. On the basis of these findings, alpha(2)M emerges as the first serum protein that can physically associate with, and thereby regulate, PP14. Moreover, this represents the first documented interaction between the protein carrier alpha(2)M and a lipocalin protein. PMID- 11023838 TI - Purified recombinant insulin-degrading enzyme degrades amyloid beta-protein but does not promote its oligomerization. AB - Amyloid beta-protein (Abeta) has been implicated as an early and essential factor in the pathogenesis of Alzheimer's disease. Although its cellular production has been studied extensively, little is known about Abeta clearance. Recently, insulin-degrading enzyme (IDE), a 110-kDa metalloendopeptidase, was found to degrade both endogenously secreted and synthetic Abeta peptides. Surprisingly, IDE-mediated proteolysis of [(125)I]Abeta(1-40) in microglial cell-culture media was accompanied by the formation of (125)I-labelled peptides with higher apparent molecular masses, raising the possibility that the degradation products act as 'seeds' for Abeta oligomerization. To directly address the role of IDE in Abeta degradation and oligomerization, we investigated the action of purified recombinant wild-type and catalytically inactive IDEs. Our data demonstrate that (i) IDE alone is sufficient to cleave purified Abeta that is either unlabelled, iodinated or (35)S-labelled; (ii) the initial cleavage sites are His(14)-Gln(15), Phe(19)-Phe(20) and Phe(20)-Ala(21); and (iii) incubation of IDE with [(125)I]Abeta, but not with [(35)S]-Abeta, leads to the formation of slower migrating species on gels. Since iodination labels N-terminal fragments of Abeta, and (35)S labels C-terminal products, we analysed unlabelled synthetic fragments of Abeta and determined that only the N-terminal fragments migrate with anomalously high molecular mass. These results indicate that IDE alone is sufficient to degrade Abeta at specific sites, and that its degradation products do not promote oligomerization of the intact Abeta peptide. PMID- 11023839 TI - The biochemical characterization of aggrecan from normal and tibial dyschondroplastic chicken growth-plate cartilage. AB - Tibial dyschondroplasia (TD) is a disorder of endochondral ossification characterized by the presence of an avascular, non-mineralized cartilage lesion extending from the growth plate into the metaphysis. Cells within the TD growth plate fail to differentiate to full hypertrophy, and instead appear to maintain a 'pre-hypertrophic' or 'transitional' status. Studies of the expression and distribution of cartilage matrix macromolecules in the TD growth plate have shown a marked decrease in the levels of aggrecan in the TD matrix. In the present study we compared the biochemical characteristics of the aggrecan molecules extracted from normal epiphyseal and TD cartilage. We have shown three major differences between normal and TD cartilage aggrecan. These are: (1) increase in molecular mass; (2) increase in the number of keratan sulphate chains; and (3) difference in the pattern of sulphation in TD aggrecan. Such changes in biochemical characteristics of the aggrecan monomers in TD cartilage may be associated with the lack of mineralization of the diseased cartilage. The present study provides a basis for further investigations into the importance of proteoglycans in normal and pathological bone development. PMID- 11023840 TI - Proteins of the endoplasmic-reticulum-associated degradation pathway: domain detection and function prediction. AB - Sequence database searches, using iterative-profile and Hidden-Markov-model approaches, were used to detect hitherto-undetected homologues of proteins that regulate the endoplasmic reticulum (ER)-associated degradation pathway. The translocon-associated subunit Sec63p (Sec=secretory) was shown to contain a domain of unknown function found twice in several Brr2p-like RNA helicases (Brr2=bad response to refrigeration 2). Additionally, Cue1p (Cue=coupling of ubiquitin conjugation to ER degradation), a yeast protein that recruits the ubiquitin-conjugating (UBC) enzyme Ubc7p to an ER-associated complex, was found to be one of a large family of putative scaffolding-domain-containing proteins that include the autocrine motility factor receptor and fungal Vps9p (Vps=vacuolar protein sorting). Two other yeast translocon-associated molecules, Sec72p and Hrd3p (Hrd=3-hydroxy-3-methylglutaryl-CoA reductase degradation), were shown to contain multiple tetratricopeptide-repeat-like sequences. From this observation it is suggested that Sec72p associates with a heat-shock protein, Hsp70, in a manner analogous to that known for Hop (Hsp70/Hsp90 organizing protein). Finally, the luminal portion of Ire1p (Ire=high inositol-requiring), thought to convey the sensing function of this transmembrane kinase and endoribonuclease, was shown to contain repeats similar to those in beta-propeller proteins. This finding hints at the mechanism by which Ire1p may sense extended unfolded proteins at the expense of compact folded molecules. PMID- 11023842 TI - Auditory symmetry analysis. AB - The study of biological symmetry continues to be an important and active area of research, yet in the hearing sciences there are no established quantitative methods for measuring auditory asymmetries and dissimilarities in threshold tuning curves (i.e. audiograms). Using a paired design and adopting methods from the analysis of fluctuating asymmetry, we describe methods for auditory researchers interested in delineating auditory asymmetries and comparing tuning curves, behavioral or neural. We illustrate the methods using audiograms of the prothoracic T-cell interneuron in a nocturnal katydid (Neoconocephalus ensiger). The results show that 87-92 % of T-cells had right-minus-left threshold asymmetries no larger than expected from measurement error alone. Thus, apart from small random fluctuating asymmetries, T-cell pairs in N. ensiger showed no sensory bias and were bilaterally symmetrical from 5 to 100 kHz. The sensitivity of the methods for detecting tuning curve dissimilarities was confirmed in a sound lateralization paradigm by comparing the 'symmetry' (i.e. similarity) of T cell tuning curves measured at 0 degrees stimulation with tuning curves measured at 90 degrees stimulation for the same T-cell. The results show that T-cell thresholds measured frontally (0 degrees ) were significantly higher than those measured laterally (90 degrees ), particularly for ultrasonic frequencies. Statistically, the directional shift (increase) in auditory thresholds was detected as a directional asymmetry in T-cell tuning, whose origin and functional significance to an insect behaving normally are discussed. The paper discusses practical considerations for detecting auditory asymmetries and tuning curve dissimilarities in general, and closes by questioning the relevance of auditory symmetry for sound localization in both vertebrates and insects. PMID- 11023841 TI - Identification of a novel 45 kDa protein (JP-45) from rabbit sarcoplasmic reticulum junctional-face membrane. AB - Using a biochemical/immunological approach to analyse the protein constituents of skeletal-muscle junctional-face membrane (JFM), we identified a 45 kDa protein. Its N-terminal amino acid was blocked, but the amino acid sequence obtained from several peptides after proteolytic treatment did not significantly match that of any protein present in the SwissProt and NCBI (National Center for Biotechnology Information) databases. We synthesized a peptide whose sequence matched that of one of the peptides obtained after CNBr cleavage of the 45 kDa protein; the peptide was conjugated to a carrier and used to raise antibodies. The antiserum was used to study in more detail the biochemical characteristics of the novel 45 kDa protein. Analysis of the proteins present in different subcellular membrane fractions show that the novel 45 kDa polypeptide: (i) is an integral membrane constituent present both in neonatal and adult skeletal-muscle sarcoplasmic reticulum; (ii) is selectively localized in the JFM; (iii) is not present in microsomes obtained from rabbit heart, liver or kidney. Immunoprecitation with anti-(45 kDa protein) antibody indicates that the 45 kDa protein is part of a complex which can be phosphorylated in vitro by the catalytic subunit of protein kinase A. PMID- 11023843 TI - Neuroethology of the katydid T-cell. I. Tuning and responses to pure tones. AB - The tuning and pure-tone physiology of the T-cell prothoracic auditory interneuron were investigated in the nocturnal katydid Neoconocephalus ensiger. The T-cell is extremely sensitive and broadly tuned, particularly to high frequency ultrasound (>20 kHz). Adult thresholds were lowest and showed their least variability for frequencies ranging from 25 to 80 kHz. The average best threshold of the T-cell in N. ensiger ranged from 28 to 38 dB SPL and the best frequency from 20 to 27 kHz. In females, the T-cell is slightly more sensitive to the range of frequencies encompassing the spectrum of male song. Tuning of the T cell in non-volant nymphs was comparable with that of adults, and this precocious ultrasound sensitivity supports the view that it has a role in the detection of terrestrial sources of predaceous ultrasound. In adults, T-cell tuning is narrower than that of the whole auditory (tympanic) organ, but only at audio frequencies. Superthreshold physiological experiments revealed that T-cell responses were ultrasound-biased, with intensity/response functions steeper and spike latencies shorter at 20, 30 and 40 kHz than at 5, 10 and 15 kHz. The same was also true for T-cell stimulation at 90 degrees compared with stimulation at 0 degrees within a frequency, which supports early T-cell research showing that excitation of the contralateral ear inhibits ipsilateral T-cell responses. In a temporal summation experiment, the integration time of the T-cell at 40 kHz (integration time constant tau =6.1 ms) was less than half that measured at 15 kHz ( tau =15.0 ms). Moreover, T-cell spiking in response to short-duration pure tone trains mimicking calling conspecifics (15 kHz) and bat echolocation hunting sequences (40 kHz) revealed that temporal pattern-copying was superior for ultrasonic stimulation. Apparently, T-cell responses are reduced or inhibited by stimulation with audio frequencies, which leads to the prediction that the T-cell will encode conspecific song less well than bat-like frequency-modulated sweeps during acoustic playback. The fact that the T-cell is one of the most sensitive ultrasound neurons in tympanate insects is most consistent with it serving an alarm, warning or escape function in both volant and non-volant katydids (nymphs and adults). PMID- 11023844 TI - Neuroethology of the katydid T-cell. II. Responses to acoustic playback of conspecific and predatory signals. AB - Although early work on the tettigoniid T large fiber suggested that it might mediate early-warning and escape behavior in katydids, the majority of research thereafter has focused on the ability of the T-cell to detect, localize and/or discriminate mate-calling song. Interestingly, T-cell responses to conspecific song are rarely examined for more than a few seconds, despite the fact that many katydids sing for minutes or hours at a time. In this paper, the second of a pair examining the physiology of the T-cell in Neoconocephalus ensiger, we recorded T cell responses using longer-duration playbacks (3 min) of conspecific song (Katydid signal 30 ms syllables, 9-25 kHz bandwidth, 12-15 kHz peak frequency) and two types of bat-like ultrasound, a 10 ms, 80->30 kHz frequency-modulated sweep (Bat 10 signal) and a 30 ms, 80->30 kHz frequency-modulated sweep (Bat 30 signal). Spiking responses were distinctly biased towards the short-duration ultrasonic signal, with more spikes per pulse, at a shorter spike latency and at a higher instantaneous firing frequency to the Bat 10 signal than to the Katydid signal or, surprisingly, to the Bat 30 signal. The ability of the T-cell to encode the temporal pattern of the stimulus was particularly striking. Only for the predatory bat signals did T-cell spiking faithfully copy the stimulus; playbacks of conspecific song resulted in significantly weaker spiking responses, particularly in male katydids. The results demonstrate that responses from the T cell alone may be sufficient for katydids to discriminate biologically relevant signals pertinent to the phonotactic behavior patterns involved in mate attraction and predator avoidance. PMID- 11023845 TI - The mechanics of prey prehension in chameleons. AB - Iguanian lizards generally use their tongue to capture prey. Because lingual prehension is based on surface phenomena (wet adhesion, interlocking), the maximal prey size that can be captured is small. However, published records show that prey items eaten by chameleons include small vertebrates such as lizards and birds, indicating that these lizards are using a different prey prehension mechanism. Using high-speed video recordings, cineradiography, electromyography, nerve transection and stimulation experiments, we investigated the function of the tongue during prey capture. The results of these experiments indicate that chameleons have modified the primitive iguanian system by including a suction component in their prehension mechanism. Suction is generated by the activity of two modified intrinsic tongue muscles that pull the tongue pad inwards. Moreover, we demonstrate that the mechanism described here is a prerequisite for successful feeding. PMID- 11023846 TI - Respiration and heart rate at the surface between dives in northern elephant seals. AB - All underwater activities of diving mammals are constrained by the need for surface gas exchange. Our aim was to measure respiratory rate (fb) and heart rate (fh) at the surface between dives in free-ranging northern elephant seals Mirounga angustirostris. We recorded fb and fh acoustically in six translocated juveniles, 1.8-2. 4 years old, and three migrating adult males from the rookery at Ano Nuevo, California, USA. To each seal, we attached a diving instrument to record the diving pattern, a satellite tag to track movements and location, a digital audio tape recorder or acoustic datalogger with an external hydrophone to record the sounds of respiration and fh at the surface, and a VHF transmitter to facilitate recovery. During surface intervals averaging 2.2+/-0.4 min, adult males breathed a mean of 32.7+/-5.4 times at a rate of 15. 3+/-1.8 breaths min( )(1) (means +/- s.d., N=57). Mean fh at the surface was 84+/-3 beats min(-)(1). The fb of juveniles was 26 % faster than that of adult males, averaging 19.2+/ 2.2 breaths min(-)(1) for a mean total of 41.2+/-5.0 breaths during surface intervals lasting 2.6+/-0.31 min. Mean fh at the surface was 106+/-3 beats min( )(1). fb and fh did not change significantly over the course of surface intervals. Surface fb and fh were not clearly associated with levels of exertion, such as rapid horizontal transit or apparent foraging, or with measures of immediately previous or subsequent diving performance, such as diving duration, diving depth or swimming speed. Together, surface respiration rate and the duration of the preceding dive were significant predictors of surface interval duration. This implies that elephant seals minimize surface time spent loading oxygen depending on rates of oxygen uptake and previous depletion of stores. PMID- 11023847 TI - Sub-ice foraging behavior of emperor penguins. AB - Emperor penguins (Aptenodytes forsteri) were equipped with a remote underwater video camera, the Crittercam, to evaluate sub-ice foraging behavior while the birds dived from an isolated dive hole. Three birds dived and foraged successfully for 1 h periods after being trained to wear and to dive with a harness for camera attachment. Video and depth profile recordings revealed that emperor penguins travel at shallow depths (<50 m), ascend to the undersurface of the ice to feed on fish, and descend back to depth to return to the exit hole. Although the mean durations of dives of individual birds with the Crittercam were 21-35 % shorter than the diving durations of these same birds without the camera, the dive profiles in both situations were similar, thus demonstrating a similar foraging strategy in birds diving without the camera. Despite shorter diving durations with the camera, the penguins were still successful at prey capture in 80 % of 91 dives greater than 1 min in duration. Prey included the sub-ice fish Pagothenia borchgrevinki. Hunting ascents (from depth to within 5 m of the surface) occurred in 85 % of dives, ranged from zero to three per dive, and were associated with successful prey capture in 77 % of 128 ascents. Occasionally, several fish were captured during a single ascent. These observations and this application of video technology create a model for further physiological and behavioral studies of foraging, and also emphasize the potential importance of shallow dives as sources of food intake for emperor penguins during foraging trips to sea. PMID- 11023848 TI - Electroreception in Gymnotus carapo: pre-receptor processing and the distribution of electroreceptor types. AB - This paper describes the peripheral mechanisms involved in signal processing of self- and conspecific-generated electric fields by the electric fish Gymnotus carapo. The distribution of the different types of tuberous electroreceptor and the occurrence of particular electric field patterns close to the body of the fish were studied. The density of tuberous electroreceptors was found to be maximal on the jaw (foveal region) and very high on the dorsal region of the snout (parafoveal region), decaying caudally. Tuberous type II electroreceptors were much more abundant than type I electroreceptors. Type I electroreceptors occurred exclusively on the head and rostral trunk regions, while type II electroreceptors were found along as much as 90 % of the fish. Electrophysiological data indicated that conspecific- and self-generated electric currents are 'funnelled' by the high conductivity and geometry of the body of the fish. These currents are concentrated at the peri-oral zone, where most electroreceptors are located. Moreover, within this region, field vector directions were collimated, constituting the most efficient stimulus for electroreceptors. It can be concluded that the passive properties of the fish tissue represent a pre-receptor device that enhances exafferent and reafferent electrical signals at the fovea-parafoveal region. PMID- 11023849 TI - Detection of bright and dim colours by honeybees. AB - Honeybees, Apis mellifera, were trained to detect coloured disks with either a strong or a weak intensity difference against the background. Green, blue, ultraviolet-reflecting white and grey papers were reciprocally combined as targets or backgrounds, providing strong chromatic and/or achromatic cues. The behavioural performance of the honeybees was always symmetrical for both reciprocal target/background combinations of a colour pair, thus showing that target detection is independent of whether the colour is presented as a background or as a target in combination with the other colour. Bright targets against dim backgrounds and vice versa were detected more reliably than dim target/background combinations. This result favours the general assumption that the detectability of a coloured stimulus increases with increasing intensity. PMID- 11023850 TI - Molecular cloning and hormonal control in the ovary of connexin 31.5 mRNA and correlation with the appearance of oocyte maturational competence in red seabream. AB - Gap junctions are aggregates of intercellular channels, composed of the protein connexin (Cx), between adjacent cells. This study examined whether, in the ovary of the red seabream Pagrus major, the connexin gene essential for the production of RNA and protein during the acquisition of oocyte maturational competence is active. Mixed primers for this reaction were designed on the basis of the high sequence homology of selected regions of known connexin genes. Polymerase-chain reaction-amplified cDNA fragments generated by 3' and 5' rapid amplication of cDNA ends were combined to generate full-length cDNA sequences. The resulting 2400 base pair cDNA had an open reading frame encoding a polypeptide containing 275 amino acid residues (31493 Da; Cx31.5). Hydropathicity analysis of the predicted amino acid sequence indicated that red seabream Cx31.5 has four major hydrophobic regions and four major hydrophilic regions indicative of a topology similar to that of known connexins. Typical connexin consensus sequences were also observed in the first and second extracellular loops. During the acquisition of oocyte maturational competence, red seabream Cx31.5 mRNA transcription levels increased after treatment with gonadotropin-II. It is therefore proposed that expression of Cx31.5 contributes to the acquisition of oocyte maturational competence in this species. PMID- 11023851 TI - The behavioral detection of binary mixtures of amino acids and their individual components by catfish. AB - The question of whether a binary mixture of amino acids is detected by fish as a unique odor or whether the qualities of the individual components are retained within the mixture was investigated in channel (Ictalurus punctatus) and brown bullhead (Ameiurus nebulosus) catfish, species that are highly similar in their olfactory receptor and behavioral responses to amino acid odorants. Catfish respond with greater appetitive food-searching (swimming) behavior to amino-acid conditioned olfactory stimuli than to non-conditioned amino acids. In the present study, appetitive food-searching behavior was measured by counting the number of turns of the fish greater than 90 degrees within 90 s of stimulus onset and, in some tests, by video tracking. The two methods yielded highly correlated results. Channel catfish conditioned to a binary mixture composed of equimolar amino acids responded with searching behavior to the amino acid that produced the larger amplitude electro-olfactogram (EOG) response as they did to the conditioned stimulus. In further studies, bullhead catfish were conditioned either to a binary mixture or to a single amino acid and tested to determine whether a binary mixture was detected as the component evoking the larger EOG response. In all initial tests (trials 1-3), the more stimulatory component of a binary mixture was not discriminated from the binary mixture; however, the less stimulatory component and all other amino acids tested were discriminated from the mixture. By increasing the concentration of the originally less potent component in a binary mixture, making it the more stimulatory compound, it was now detected as not significantly different from the binary mixture; however, the original more potent component (i.e. now the less potent stimulus) was detected as significantly different from the mixture. However, with 5-10 additional discrimination training trials, the less stimulatory component in a binary mixture influenced the perception of the binary mixture because the binary mixture was no longer detected only as its more stimulatory component. The data suggest that a two-step learning process occurs within the olfactory bulb and possibly higher-order telencephalic nuclei. PMID- 11023852 TI - Effects of body size on take-off flight performance in the Phasianidae (Aves). AB - To evaluate the mechanisms responsible for relationships between body mass and maximum take-off performance in birds, we studied four species in the Phasianidae: northern bobwhite (Colinus virginianus), chukar (Alectoris chukar), ring-necked pheasant (Phasianus colchicus) and wild turkey (Meleagris gallopavo). These species vary in body mass from 0.2 to 5.3 kg, and they use flight almost solely to escape predators. During take-off, all the species used a similar wingbeat style that appeared to be a vortex-ring gait with a tip reversal during the upstroke. The tip reversal is unusual for birds with rounded wings; it may offer an aerodynamic advantage during rapid acceleration. Flight anatomy generally scaled geometrically, except for average wing chord and wing area, which increased more than expected as body mass (m) increased. Pectoralis strain varied from 19.1 to 35.2 % and scaled in proportion to m(0.23). This positive scaling is not consistent with the widely held assumption that muscle strain is independent of body mass among geometrically similar species. The anatomy of the species precluded measurements of in vivo pectoralis force using the strain-gauge technique that has been employed successfully in other bird species, so we could not directly test in vivo pectoralis force-velocity relationships. However, whole body kinematics revealed that take-off power (P(ta)), the excess power available for climbing and accelerating in flight, scaled in proportion to m(0.75) and that pectoralis mass-specific P(ta) decreased in proportion to m(-)(0.26) and was directly proportional to wingbeat frequency. These trends suggest that mass specific pectoralis work did not vary with body mass and that pectoralis stress and strain were inversely proportional, as expected from classical force-velocity models for skeletal muscle. Our observations of P(ta) were consistent with evidence from other species engaged in escape flight and, therefore, appear to contradict evidence from studies of take-off or hovering with an added payload. PMID- 11023853 TI - Seasonal changes in energy and water use by verdins, Auriparus flaviceps. AB - We used the doubly labeled water (DLW) method to measure field metabolic rate (FMR) and water turnover during winter and summer in a very small (6.5 g) insectivorous desert passerine bird, the verdin (Auriparus flaviceps). Concurrently, we monitored weather conditions and used time-activity budget data of free-living birds and laboratory data on resting metabolic rate to construct time-activity laboratory (TAL) estimates of daily energy expenditure (DEE ) and to partition the verdins' energy budget into thermoregulatory, activity and basal components. The FMR of adult verdins was 33.6+/-0.9 kJ day(-)(1) (mean +/- s.e.m.; N=14) in winter and 22.8+/-0.45 kJ day(-)(1) (N=7) in summer. FMR correlated negatively with the mean standard operative environmental temperature (T(es)) prevailing during the measurement period. TAL analysis produced DEE estimates that corresponded on average to within -0.9+/-2.4 % of our DLW-measured FMR values (range of individual values -18.3 % to +14.3 %). Metabolic expeditures for cold defense were 19.5+/-2.1 % of DEE in winter (computed assuming substitution of exercise thermogenesis for thermoregulatory costs in active birds). In the summer, thermoregulatory costs amounted to 9.0+/-0.4 % of DEE for keeping warm and 1.0+/-0.1 % of DEE for keeping cool in the heat. Activity costs were 21.0+/-0.5 % of DEE in winter and 17.5+/-0.1 % of DEE in summer. The water efflux of nonbreeding adult verdins was 3.9+/-0.2 ml day(-)(1) (624+/-22 ml kg( )(1 )day(-)(1)) in summer (N=5) and 3. 4+/-0.2 ml day(-)(1) (498+/-26 ml kg(-)(1 )day(-)(1)) in winter (N=14). The water economy index (WEI; water efflux per unit FMR) of verdins was higher in summer (0.17+/-0.01 ml kJ(-)(1)) than in winter (0.10+/-0.01 ml kJ(-)(1)) and correlated negatively with mean T(es). PMID- 11023854 TI - Bioconvective dynamics: dependence on organism behaviour. AB - Bioconvection occurs when a macroscopic nonuniformity of the concentration of microbial populations is generated and maintained by the directional swimming of the organisms. This study investigated the properties of the patterns near the onset of the instability and later during its evolution into a fully nonlinear convection regime. In suspensions of the bacteria Bacillus subtilis, which tend to swim upwards in a gradient of oxygen concentration that they create by consumption, we discovered that the dominant wavelength at the onset of the instability is determined primarily by the cell density and is influenced only weakly by the fluid depth. This observation contrasts strongly with previous observations on the gravitactic alga Chlamydomonas nivalis, in which the opposite dependence was found. Considerable differences were also found in the long-term evolution of the convection patterns. These results demonstrate the existence of readily distinguishable types of bioconvection systems, even at early stages of the instability. The observed differences are clearly and causally correlated with disparate reasons for upward swimming by these micro-organisms, leading to different geometric distributions of the density of the suspension. PMID- 11023855 TI - Mitochondrial oxyconformity and cold adaptation in the polychaete Nereis pelagica and the bivalve Arctica islandica from the Baltic and White Seas. AB - The rates of oxygen uptake of the marine polychaete Nereis pelagica and the bivalve Arctica islandica depend on the availability of ambient oxygen. This is manifest both at the tissue level and in isolated mitochondria studied between oxygen tensions (P(O2)) of 6.3 and 47.6 kPa (47-357 mmHg). Oxyconformity was found in both Baltic Sea (Kiel Bight) and cold-adapted White Sea populations of the two species. However, mitochondria isolated from White Sea specimens of N. pelagica and A. islandica showed a two- to threefold higher aerobic capacity than mitochondria prepared from Baltic Sea specimens. We tested whether mitochondrial oxyconformity can be explained by an additional electron pathway that is directly controlled by P(O2). Mitochondrial respiration of both invertebrate species was inhibited by cyanide (KCN) and by salicylhydroxamic acid (SHAM). The overall rate of mitochondrial oxygen consumption increased at high P(O2). Phosphorylation efficiency (ADP/O ratio) decreased at elevated P(O2) (27.5-47.6 kPa, 206-357 mmHg), regardless of whether malate or succinate was used as a substrate. In contrast to the invertebrate mitochondria studied, mitochondria isolated from bovine heart, as an oxyregulating control species, did not show an elevated rate of oxygen uptake at high P(O2) in any respiratory state, with the exception of state 2 malate respiration. In addition, rates of ATP formation, respiratory control ratios (RCR) and ADP/O ratios remained virtually unchanged or even tended to decreased. In conclusion, the comparison between mitochondria from oxyregulating and oxyconforming organisms supports the existence of an alternative oxidase in addition to the classical cytochrome c oxidase. In accordance with models discussed previously, oxidative phosphorylation does not explain the rate of mitochondrial oxygen consumption during progressive activation of the alternative electron transport system. We discuss the alternative system, thought to be adaptive in confined, usually hypoxic environments, where excess oxygen can be eliminated and oxygen levels can be kept low by an increase in the rate of oxygen consumption, thereby minimizing the risk of oxidative stress. PMID- 11023856 TI - A human YAC transgene rescues craniofacial and neural tube development in PDGFRalpha knockout mice and uncovers a role for PDGFRalpha in prenatal lung growth. AB - The platelet-derived growth factor alpha-receptor (PDGFRalpha) plays a vital role in the development of vertebrate embryos, since mice lacking PDGFRalpha die in mid-gestation. PDGFRalpha is expressed in several types of migratory progenitor cells in the embryo including cranial neural crest cells, lung smooth muscle progenitors and oligodendrocyte progenitors. To study PDGFRalpha gene regulation and function during development, we generated transgenic mice by pronuclear injection of a 380 kb yeast artificial chromosome (YAC) containing the human PDGFRalpha gene. The YAC transgene was expressed in neural crest cells, rescued the profound craniofacial abnormalities and spina bifida observed in PDGFRalpha knockout mice and prolonged survival until birth. The ultimate cause of death was respiratory failure due to a defect in lung growth, stemming from failure of the transgene to be expressed correctly in lung smooth muscle progenitors. However, the YAC transgene was expressed faithfully in oligodendrocyte progenitors, which was not previously observed with plasmid-based transgenes containing only upstream PDGFRalpha control sequences. Our data illustrate the complexity of PDGFRalpha genetic control, provide clues to the location of critical regulatory elements and reveal a requirement for PDGF signalling in prenatal lung growth, which is distinct from the known requirement in postnatal alveogenesis. In addition, we found that the YAC transgene did not prolong survival of Patch mutant mice, indicating that genetic defects outside the PDGFRalpha locus contribute to the early embryonic lethality of Patch mice. PMID- 11023857 TI - Brain derived neurotrophic factor is an endothelial cell survival factor required for intramyocardial vessel stabilization. AB - Brain derived neurotrophic factor, BDNF, is a neurotrophin best characterized for its survival and differentiative effects on neurons expressing the trk B receptor tyrosine kinase. Although many of these neurons are lost in the BDNF(-)(/)(- )mouse, the early postnatal lethality of these animals suggests a wider function for this growth factor. Here, we demonstrate that deficient expression of BDNF impairs the survival of endothelial cells in intramyocardial arteries and capillaries in the early postnatal period, although the embryonic vasculature can remodel into arteries, capillaries and veins. BDNF deficiency results in a reduction in endothelial cell-cell contacts and in endothelial cell apoptosis, leading to intraventricular wall hemorrhage, depressed cardiac contractility and early postnatal death. Vascular hemorrhage is restricted to cardiac vessels, reflecting the localized expression of BDNF and trk B by capillaries and arterioles in this vascular bed. Conversely, ectopic BDNF overexpression in midgestational mouse hearts results in an increase in capillary density. Moreover, BDNF activation of endogenous trk B receptors supports the survival of cardiac microvascular endothelial cells cultured from neonatal mice. These results establish an essential role for BDNF in maintaining vessel stability in the heart through direct angiogenic actions on endothelial cells. PMID- 11023858 TI - Nitric oxide and cGMP influence axonogenesis of antennal pioneer neurons. AB - The grasshopper embryo has been used as a convenient system with which to investigate mechanisms of axonal navigation and pathway formation at the level of individual nerve cells. Here, we focus on the developing antenna of the grasshopper embryo (Schistocerca gregaria) where two siblings of pioneer neurons establish the first two axonal pathways to the CNS. Using immunocytochemistry we detected nitric oxide (NO)-induced synthesis of cGMP in the pioneer neurons of the embryonic antenna. A potential source of NO are NADPH-diaphorase-stained epithelial cells close to the basal lamina. To investigate the role of the NO/cGMP signaling system during pathfinding, we examined the pattern of outgrowing pioneer neurons in embryo culture. Pharmacological inhibition of soluble guanylyl cyclase (sGC) and of NO synthase (NOS) resulted in an abnormal pattern of pathway formation in the antenna. Axonogenesis of both pairs of pioneers was inhibited when specific NOS or sGC inhibitors were added to the culture medium; the observed effects include the loss axon emergence as well as retardation of outgrowth, such that growth cones do not reach the CNS. The addition of membrane-permeant cGMP or a direct activator of the sGC enzyme to the culture medium completely rescued the phenotype resulting from the block of NO/cGMP signaling. These results indicate that NO/cGMP signaling is involved in axonal elongation of pioneer neurons in the antenna of the grasshopper. PMID- 11023859 TI - Hes1 is a negative regulator of inner ear hair cell differentiation. AB - Hair cell fate determination in the inner ear has been shown to be controlled by specific genes. Recent loss-of-function and gain-of-function experiments have demonstrated that Math1, a mouse homolog of the Drosophila gene atonal, is essential for the production of hair cells. To identify genes that may interact with Math1 and inhibit hair cell differentiation, we have focused on Hes1, a mammalian hairy and enhancer of split homolog, which is a negative regulator of neurogenesis. We report here that targeted deletion of Hes1 leads to formation of supernumerary hair cells in the cochlea and utricle of the inner ear. RT-PCR analysis shows that Hes1 is expressed in inner ear during hair cell differentiation and its expression is maintained in adulthood. In situ hybridization with late embryonic inner ear tissue reveals that Hes1 is expressed in supporting cells, but not hair cells, of the vestibular sensory epithelium. In the cochlea, Hes1 is selectively expressed in the greater epithelial ridge and lesser epithelial ridge regions which are adjacent to inner and outer hair cells. Co-transfection experiments in postnatal rat explant cultures show that overexpression of Hes1 prevents hair cell differentiation induced by Math1. Therefore Hes1 can negatively regulate hair cell differentiation by antagonizing Math1. These results suggest that a balance between Math1 and negative regulators such as Hes1 is crucial for the production of an appropriate number of inner ear hair cells. PMID- 11023860 TI - Cell interactions within nascent neural crest cell populations transiently promote death of neurogenic precursors. AB - We have previously shown that cultured trunk neural crest cell populations irreversibly lose neurogenic ability when dispersal is prevented or delayed, while the ability to produce other crest derivatives is retained (Vogel, K. S. and Weston, J. A. (1988) Neuron 1, 569-577). Here, we show that when crest cells are prevented from dispersing, cell death is increased and neurogenesis is decreased in the population, as a result of high cell density. Control experiments to characterize the effects of high cell density on environmental conditions in culture suggest that reduced neurogenesis is the result of cell cell interactions and not changes (conditioning or depletion) of the culture medium. Additionally, we show that the caspase inhibitor zVAD-fmk, which blocks developmentally regulated cell death, rescues the neurogenic ability of high density cultures, without any apparent effect on normal, low-density cultures. We conclude, therefore, that increased cell interaction at high cell densities results in the selective death of neurogenic precursors in the nascent crest population. Furthermore, we show that neurogenic cells in cultured crest cell populations that have dispersed immediately are not susceptible to contact mediated death, even if they are subsequently cultured at high cell density. Since most early migrating avian crest cells express Notch1, and a subset expresses Delta1 (Wakamatsu, Y., Maynard, T. M. and Weston, J. A. (2000) Development 127, 2811-2821), we tested the possibility that the effects of cell contact were mediated by components of a Notch signaling pathway. We found that neurogenic precursors are eliminated when crest cells are co-cultured with exogenous Delta1-expressing cells immediately after they segregate from the neural tube, although not after they have previously dispersed. We conclude that early and prolonged cell interactions, mediated at least in part by Notch signaling, can regulate the survival of neurogenic cells within the nascent crest population. We suggest that a transient episode of cell contact-mediated death of neurogenic cells may serve to eliminate fate-restricted neurogenic cells that fail to disperse promptly in vivo. PMID- 11023861 TI - NARROW SHEATH1 functions from two meristematic foci during founder-cell recruitment in maize leaf development. AB - The narrow sheath duplicate genes (ns1 and ns2) perform redundant functions during maize leaf development. Plants homozygous for mutations in both ns genes fail to develop wild-type leaf tissue in a lateral domain that includes the leaf margin. Previous studies indicated that the NS gene product(s) functions during recruitment of leaf founder-cells in a lateral, meristematic domain that contributes to leaf margin development. A mosaic analysis was performed in which the ns1-O mutation was exposed in hemizygous, clonal sectors in a genetic background already homozygous for ns2-O. Analyses of mutant, sectored plants demonstrate that NS1 function is required in L2-derived tissue layers for development of the narrow sheath leaf domain. NS1 function is not required for development of the central region of maize leaves. Furthermore, the presence of the non-mutant ns1 gene outside the narrow sheath domain cannot compensate for the absence of the non-mutant gene within the narrow sheath domain. NS1 acts non cell autonomously within the narrow sheath-margin domain and directs recruitment of marginal, leaf founder cells from two discrete foci in the maize meristem. Loss of NS1 function during later stages of leaf development results in no phenotypic consequences. These data support our model for NS function during founder-cell recruitment in the maize meristem. PMID- 11023862 TI - Multiple levels of regulation specify the polarity of an asymmetric cell division in C. elegans. AB - Wnt signaling systems play important roles in the generation of cell and tissue polarity during development. We describe a Wnt signaling system that acts in a new way to orient the polarity of an epidermal cell division in C. elegans. In this system, the EGL-20/Wnt signal acts in a permissive fashion to polarize the asymmetric division of a cell called V5. EGL-20 regulates this polarization by counteracting lateral signals from neighboring cells that would otherwise reverse the polarity of the V5 cell division. Our findings indicate that this lateral signaling pathway also involves Wnt pathway components. Overexpression of EGL-20 disrupts both the asymmetry and polarity of lateral epidermal cell divisions all along the anteroposterior (A/P) body axis. Together our findings suggest that multiple, inter-related Wnt signaling systems may act together to polarize asymmetric cell divisions in this tissue. PMID- 11023864 TI - Mediolateral somitic origin of ribs and dermis determined by quail-chick chimeras. AB - Somites are transient mesodermal structures giving rise to all skeletal muscles of the body, the axial skeleton and the dermis of the back. Somites arise from successive segmentation of the presomitic mesoderm (PSM). They appear first as epithelial spheres that rapidly differentiate into a ventral mesenchyme, the sclerotome, and a dorsal epithelial dermomyotome. The sclerotome gives rise to vertebrae and ribs while the dermomyotome is the source of all skeletal muscles and the dorsal dermis. Quail-chick fate mapping and diI-labeling experiments have demonstrated that the epithelial somite can be further subdivided into a medial and a lateral moiety. These two subdomains are derived from different regions of the primitive streak and give rise to different sets of muscles. The lateral somitic cells migrate to form the musculature of the limbs and body wall, known as the hypaxial muscles, while the medial somite gives rise to the vertebrae and the associated epaxial muscles. The respective contribution of the medial and lateral somitic compartments to the other somitic derivatives, namely the dermis and the ribs has not been addressed and therefore remains unknown. We have created quail-chick chimeras of either the medial or lateral part of the PSM to examine the origin of the dorsal dermis and the ribs. We demonstrate that the whole dorsal dermis and the proximal ribs exclusively originates from the medial somitic compartment, whereas the distal ribs derive from the lateral compartment. PMID- 11023863 TI - Extraocular mesenchyme patterns the optic vesicle during early eye development in the embryonic chick. AB - The vertebrate eye develops from the neuroepithelium of the ventral forebrain by the evagination and formation of the optic vesicle. Classical embryological studies have shown that the surrounding extraocular tissues - the surface ectoderm and extraocular mesenchyme - are necessary for normal eye growth and differentiation. We have used explant cultures of chick optic vesicles to study the regulation of retinal pigmented epithelium (RPE) patterning and differentiation during early eye development. Our results show that extraocular mesenchyme is required for the induction and maintenance of expression of the RPE specific genes Mitf and Wnt13 and the melanosomal matrix protein MMP115. In the absence of extraocular tissues, RPE development did not occur. Replacement of the extraocular mesenchyme with cranial mesenchyme, but not lateral plate mesoderm, could rescue expression of the RPE-marker Mitf. In addition to activating expression of RPE-specific genes, the extraocular mesenchyme inhibits the expression of the neural retina-specific transcription factor Chx10 and downregulates the eye-specific transcription factors Pax6 and Optx2. The TGF(&bgr;) family member activin can substitute for the extraocular mesenchyme by promoting expression of the RPE-specific genes and downregulating expression of the neural retina-specific markers. These data indicate that extraocular mesenchyme, and possibly an activin-like signal, pattern the domains of the optic vesicle into RPE and neural retina. PMID- 11023865 TI - Tissue-specific regulation of cyclin E transcription during Drosophila melanogaster embryogenesis. AB - Cyclin E is an essential regulator of S phase entry. We have previously shown that transcriptional regulation of the gene that encodes Drosophila cyclin E, DmcycE, plays an important role in the control of the G(1) to S phase transition during development. We report here the first comprehensive analysis of the transcriptional regulation of a G(1 )phase cell cycle regulatory gene during embryogenesis. Analysis of deficiencies, a genomic transformant and reporter gene constructs revealed that DmcycE transcription is controlled by a large and complex cis-regulatory region containing tissue- and stage-specific components. Separate regulatory elements for transcription in epidermal cells during cell cycles 14-16, central nervous system cells and peripheral nervous system cells were found. An additional cis-regulatory element drives transcription in thoracic epidermal cells that undergo a 17th cell cycle when other epidermal cells have arrested in G(1 )phase prior to terminal differentiation. The complexity of DmcycE transcriptional regulation argues against a model in which DmcycE transcription is regulated simply and solely by G(1) to S phase transcription regulators such as RB, E2F and DP. Rather, our study demonstrates that tissue specific transcriptional regulatory mechanisms are important components of the control of cyclin E transcription and thus of cell proliferation in metazoans. PMID- 11023866 TI - Spatial expression of Hox cluster genes in the ontogeny of a sea urchin. AB - The Hox cluster of the sea urchin Strongylocentrous purpuratus contains ten genes in a 500 kb span of the genome. Only two of these genes are expressed during embryogenesis, while all of eight genes tested are expressed during development of the adult body plan in the larval stage. We report the spatial expression during larval development of the five 'posterior' genes of the cluster: SpHox7, SpHox8, SpHox9/10, SpHox11/13a and SpHox11/13b. The five genes exhibit a dynamic, largely mesodermal program of expression. Only SpHox7 displays extensive expression within the pentameral rudiment itself. A spatially sequential and colinear arrangement of expression domains is found in the somatocoels, the paired posterior mesodermal structures that will become the adult perivisceral coeloms. No such sequential expression pattern is observed in endodermal, epidermal or neural tissues of either the larva or the presumptive juvenile sea urchin. The spatial expression patterns of the Hox genes illuminate the evolutionary process by which the pentameral echinoderm body plan emerged from a bilateral ancestor. PMID- 11023867 TI - FIGalpha, a germ cell-specific transcription factor required for ovarian follicle formation. AB - Primordial follicles are formed perinatally in mammalian ovaries and at birth represent the lifetime complement of germ cells. With cyclic periodicity, cohorts enter into a growth phase that culminates in ovulation of mature eggs, but little is known about the regulatory cascades that govern these events. FIGalpha, a transcription factor implicated in postnatal oocyte-specific gene expression, is detected as early as embryonic day 13. Mouse lines lacking FIGalpha were established by targeted mutagenesis in embryonic stem cells. Although embryonic gonadogenesis appeared normal, primordial follicles were not formed at birth, and massive depletion of oocytes resulted in shrunken ovaries and female sterility. Fig(&agr;) (the gene for FIGalpha null males have normal fertility. The additional observation that null females do not express Zp1, Zp2 or Zp3 indicates that FIGalpha plays a key regulatory role in the expression of multiple oocyte specific genes, including those that initiate folliculogenesis and those that encode the zona pellucida required for fertilization and early embryonic survival. The persistence of FIGalpha in adult females suggests that it may regulate additional pathways that are essential for normal ovarian development. PMID- 11023868 TI - Establishment of left/right asymmetry in neuroblast migration by UNC-40/DCC, UNC 73/Trio and DPY-19 proteins in C. elegans. AB - The bilateral C. elegans neuroblasts QL and QR are born in the same anterior/posterior (A/P) position, but polarize and migrate left/right asymmetrically: QL migrates toward the posterior and QR migrates toward the anterior. After their migrations, QL but not QR switches on the Hox gene mab-5. We find that the UNC-40/netrin receptor and a novel transmembrane protein DPY-19 are required to orient these cells correctly. In unc-40 or dpy-19 mutants, the Q cells polarize randomly; in fact, an individual Q cell polarizes in multiple directions over time. In addition, either cell can express MAB-5. Both UNC-40 and DPY-19, as well as the Trio/GTPase exchange factor homolog UNC-73, are required for full polarization and migration. Thus, these proteins appear to participate in a signaling system that orients and polarizes these migrating cells in a left/right asymmetrical fashion during development. The C. elegans netrin UNC-6, which guides many cells and axons along the dorsoventral axis, is not involved in Q cell polarization, suggesting that a different netrin-like ligand serves to polarize these cells along the anteroposterior axis. PMID- 11023869 TI - Molecular, biochemical and functional analysis of a novel and developmentally important fibrillar collagen (Hcol-I) in hydra. AB - The body wall of hydra (a member of the phylum Cnidaria) is structurally reduced to an epithelial bilayer with an intervening extracellular matrix (ECM). Previous studies have established that cell-ECM interactions are important for morphogenesis and cell differentiation in this simple metazoan. The ECM of hydra is particularly interesting because it represents a primordial form of matrix. Despite progress in our understanding of hydra ECM, we still know little about the nature of hydra collagens. In the current study we provide a molecular, biochemical and functional analysis of a hydra fibrillar collagen that has similarity to vertebrate type I and type II collagens. This fibrillar collagen has been named hydra collagen-I (Hcol-I) because of its structure and because it is the first ECM collagen to be identified in hydra. It represents a novel member of the collagen family. Similar to vertebrate type I and II collagens, Hcol-I contains an N-terminal propeptide-like domain, a triple helical domain containing typical Gly-X-Y repeats and a C-terminal propeptide domain. The overall identity to vertebrate fibrillar collagens is about 30%, while the identity of the C terminal propeptide domain is 50%. Because the N-terminal propeptide domain is retained after post-translational processing, Hcol-I does not form thick fibers as seen in vertebrates. This was confirmed using transmission electron microscopy to study rotary shadow images of purified Hcol-I. In addition, absence of crucial lysine residues and an overall reduction in proline content, results in reduced crosslinking of fibrils and increased flexibility of the molecule, respectively. These structural changes in Hcol-I help to explain the flexible properties of hydra ECM. Immunocytochemical studies indicate that Hcol-I forms the 10 nm fibrils that comprise the majority of molecules in the central fibrous zone of hydra ECM. The central fibrous zone resides between the two subepithelial zones where hydra laminin is localized. While previous studies have shown that basal lamina components like laminin are expressed by the endoderm, in situ hybridisation studies show that Hcol-I mRNA expression is restricted to the ectoderm. Hcol-I expression is upregulated during head regeneration, and antisense studies using thio-oligonucleotides demonstrated that blocking the translation of Hcol-I leads to a reversible inhibition of head morphogenesis during this regenerative process. Taken in total, the data presented in this study indicate that Hcol-I is required for morphogensis in hydra and represents a novel fibrillar collagen whose structural characteristics help to explain the unique biophysical properties of hydra ECM. Interestingly, the structure of Hcol I mimics what is seen in Ehlers-Danlos syndrome type VII in humans; an inherited pathological condition that leads to joint and skin abnormalities. Hcol-I therefore illustrates an adaptive trait in which the normal physiological situation in hydra translates into a pathological condition in humans. PMID- 11023870 TI - The role of the yolk syncytial layer in germ layer patterning in zebrafish. AB - Formation of the three germ layers requires a series of inductive events during early embryogenesis. Studies in zebrafish indicate that the source of these inductive signals may be the extra-embryonic yolk syncytial layer (YSL). The characterization of genes encoding the nodal-related factor, Squint, and homeodomain protein, Bozozok, both of which are expressed in the YSL, suggested that the YSL has a role in mesendoderm induction. However, these genes, and a second nodal-related factor, cyclops, are also expressed in the overlying marginal blastomeres, raising the possibility that the marginal blastomeres can induce mesendodermal genes independently of the YSL. We have developed a novel technique to study signaling from the YSL in which we specifically eliminate RNAs in the YSL, thus addressing the in vivo requirement of RNA-derived signals from this region in mesendoderm induction. We show that injection of RNase into the yolk cell after the 1K cell stage (3 hours) effectively eliminates YSL transcripts without affecting ubiquitously expressed genes in the blastoderm. We also present data that indicate the stability of existing proteins in the YSL is unaffected by RNase injection. Using this technique, we show that RNA in the YSL is required for the formation of ventrolateral mesendoderm and induction of the nodal-related genes in the ventrolateral marginal blastomeres, revealing the presence of an unidentified inducing signal released from the YSL. We also demonstrate that the dorsal mesoderm can be induced independently of signals from the YSL and present evidence that this is due to the stabilization of (&bgr;) catenin in the dorsal marginal blastomeres. Our results demonstrate that germ layer formation and patterning in zebrafish uses a combination of YSL-dependent and -independent inductive events. PMID- 11023871 TI - Edar/Eda interactions regulate enamel knot formation in tooth morphogenesis. AB - tabby and downless mutant mice have apparently identical defects in teeth, hair and sweat glands. Recently, genes responsible for these spontaneous mutations have been identified. downless (Dl) encodes Edar, a novel member of the tumour necrosis factor (TNF) receptor family, containing the characteristic extracellular cysteine rich fold, a single transmembrane region and a death homology domain close to the C terminus. tabby (Ta) encodes ectodysplasin-A (Eda) a type II membrane protein of the TNF ligand family containing an internal collagen-like domain. As predicted by the similarity in adult mutant phenotype and the structure of the proteins, we demonstrate that Eda and Edar specifically interact in vitro. We have compared the expression pattern of Dl and Ta in mouse development, taking the tooth as our model system, and find that they are not expressed in adjacent cells as would have been expected. Teeth develop by a well recorded series of epithelial-mesenchymal interactions, similar to those in hair follicle and sweat gland development, the structures found to be defective in tabby and downless mice. We have analysed the downless mutant teeth in detail, and have traced the defect in cusp morphology back to initial defects in the structure of the tooth enamel knot at E13. Significantly, the defect is distinct from that of the tabby mutant. In the tabby mutant, there is a recognisable but small enamel knot, whereas in the downless mutant the knot is absent, but enamel knot cells are organised into a different shape, the enamel rope, showing altered expression of signalling factors (Shh, Fgf4, Bmp4 and Wnt10b). By adding a soluble form of Edar to tooth germs, we were able to mimic the tabby enamel knot phenotype, demonstrating the involvement of endogenous Eda in tooth development. We could not, however, reproduce the downless phenotype, suggesting the existence of yet another ligand or receptor, or of ligand-independent activation mechanisms for Edar. Changes in the structure of the enamel knot signalling centre in downless tooth germs provide functional data directly linking the enamel knot with tooth cusp morphogenesis. We also show that the Lef1 pathway, thought to be involved in these mutants, functions independently in a parallel pathway. PMID- 11023872 TI - Members of the bHLH-PAS family regulate Shh transcription in forebrain regions of the mouse CNS. AB - The secreted protein sonic hedgehog (Shh) is required to establish patterns of cellular growth and differentiation within ventral regions of the developing CNS. The expression of Shh in the two tissue sources responsible for this activity, the axial mesoderm and the ventral midline of the neural tube, is regulated along the anteroposterior neuraxis. Separate cis-acting regulatory sequences have been identified which direct Shh expression to distinct regions of the neural tube, supporting the view that multiple genes are involved in activating Shh transcription along the length of the CNS. We show here that the activity of one Shh enhancer, which directs reporter expression to portions of the ventral midbrain and diencephalon, overlaps both temporally and spatially with the expression of Sim2. Sim2 encodes a basic helix-loop-helix (bHLH-PAS) PAS domain containing transcriptional regulator whose Drosophila homolog, single-minded, is a master regulator of ventral midline development. Both vertebrate and invertebrate Sim family members were found sufficient for the activation of the Shh reporter as well as endogenous Shh mRNA. Although Shh expression is maintained in Sim2(-)(/)(-) embryos, it was determined to be absent from the rostral midbrain and caudal diencephalon of embryos carrying a dominant-negative transgene that disrupts the function of bHLH-PAS proteins. Together, these results suggest that bHLH-PAS family members are required for the regulation of Shh transcription within aspects of the ventral midbrain and diencephalon. PMID- 11023873 TI - BMP4 rescues a non-cell-autonomous function of Msx1 in tooth development. AB - The development of many organs depends on sequential epithelial-mesenchymal interactions, and the developing tooth germ provides a powerful model for elucidating the nature of these inductive tissue interactions. In Msx1-deficient mice, tooth development arrests at the bud stage when Msx1 is required for the expression of Bmp4 and Fgf3 in the dental mesenchyme (Bei, M. and Maas, R. (1998) Development 125, 4325-4333). To define the tissue requirements for Msx1 function, we performed tissue recombinations between wild-type and Msx1 mutant dental epithelium and mesenchyme. We show that through the E14.5 cap stage of tooth development, Msx1 is required in the dental mesenchyme for tooth formation. After the cap stage, however, tooth development becomes Msx1 independent, although our experiments identify a further late function of Msx1 in odontoblast and dental pulp survival. These results suggest that prior to the cap stage, the dental epithelium receives an Msx1-dependent signal from the dental mesenchyme that is necessary for tooth formation. To further test this hypothesis, Msx1 mutant tooth germs were first cultured with either BMP4 or with various FGFs for two days in vitro and then grown under the kidney capsule of syngeneic mice to permit completion of organogenesis and terminal differentiation. Previously, using an in vitro culture system, we showed that BMP4 stimulated the growth of Msx1 mutant dental epithelium (Chen, Y., Bei, M. Woo, I., Satokata, I. and Maas, R. (1996). Development 122, 3035-3044). Using the more powerful kidney capsule grafting procedure, we now show that when added to explanted Msx1-deficient tooth germs prior to grafting, BMP4 rescues Msx1 mutant tooth germs all the way to definitive stages of enamel and dentin formation. Collectively, these results establish a transient functional requirement for Msx1 in the dental mesenchyme that is almost fully supplied by BMP4 alone, and not by FGFs. In addition, they formally prove the postulated downstream relationship of BMP4 with respect to Msx1, establish the non-cell-autonomous nature of Msx1 during odontogenesis, and disclose an additional late survival function for Msx1 in odontoblasts and dental pulp. PMID- 11023874 TI - Parental origin-specific developmental defects in mice with uniparental disomy for chromosome 12. AB - Genetic analysis has shown that the distal portion of mouse chromosome 12 is imprinted; however, the developmental roles of imprinted genes in this region are not known. We have therefore generated conceptuses with uniparental disomy for chromosome 12, in which both copies of chromosome 12 are either paternally or maternally derived (pUPD12 and mUPD12, respectively). Both types of UPD12 result in embryos that are non-viable and that exhibit distinct developmental abnormalities. Embryos with pUPD12 die late in gestation, whereas embryos with mUPD12 can survive to term but die perinatally. The mUPD12 conceptuses are invariably growth-retarded while pUPD12 conceptuses exhibit placentomegaly. Skeletal muscle maturation defects are evident in both types of UPD12. In addition, embryos with paternal UPD12 have costal cartilage defects and hypo ossification of mesoderm-derived bones. In embryos with mUPD12, the development of the neural crest-derived middle ear ossicles is defective. Some of these anomalies are consistent with those seen with uniparental disomies of the orthologous chromosome 14 region in humans. Thus, imprinted genes on chromosome 12 are essential for viability, the regulation of prenatal growth, and the development of mesodermal and neural crest-derived lineages. PMID- 11023875 TI - Regulation of Drosophila wing vein patterning: net encodes a bHLH protein repressing rhomboid and is repressed by rhomboid-dependent Egfr signalling. AB - The stereotyped pattern of veins in the Drosophila wing is generated in response to local EGF signalling. Mutations in the rhomboid (rho) gene, which encodes a sevenpass membrane protein required to enhance signalling transmitted by the EGF receptor (Egfr), inhibit vein development and disrupt the vein pattern. By contrast, net mutations produce ectopic veins in intervein regions. We have cloned the net gene and show that it encodes a basic HLH protein that probably acts as a transcriptional repressor. net and rho are expressed in mutually exclusive patterns during the development of the wing imaginal disc. Lack of net activity causes rho expression to expand, and vice versa. Furthermore, ectopic expression of net or rho results in their mutual repression and thus suppresses vein formation or generates tube-like wings composed of vein-like tissue. Egfr signalling and net exert mutually antagonising activities during the specification of vein versus intervein fate. While Egfr signalling represses net transcription, net exhibits a two-tiered control by repressing rho transcription and interfering with Egfr signalling downstream of Rho. Our results further suggest that net is required to maintain intervein development by restricting Egfr signalling, which promotes vein development, to the Net-free vein regions of the wing disc. PMID- 11023876 TI - Ion channel selectivity using an electric stew. PMID- 11023877 TI - Molecular motions in Fourier transform space. PMID- 11023878 TI - Excitation transfer in the peridinin-chlorophyll-protein of Amphidinium carterae. AB - Peridinin-chlorophyll-protein (PCP) is a unique light-harvesting protein that uses carotenoids as its primary light-absorbers. This paper theoretically investigates excitation transfer between carotenoids and chlorophylls in PCP of the dinoflagellate Amphidinium carterae. Calculations based on a description of the electronic states of the participating chromophores and on the atomic level structure of PCP seek to identify the mechanism and pathways of singlet excitation flow. After light absorption the optically allowed states of peridinins share their electronic excitation in excitonic fashion, but are not coupled strongly to chlorophyll residues in PCP. Instead, a gateway to chlorophyll Q(y) excitations is furnished through a low-lying optically forbidden excited state, populated through internal conversion. Carbonyl group and non hydrogen side groups of peridinin are instrumental in achieving the respective coupling to chlorophyll. Triplet excitation transfer to peridinins, mediated by electron exchange, is found to efficiently protect chlorophylls against photo oxidation. PMID- 11023879 TI - Energy transfer among CP29 chlorophylls: calculated Forster rates and experimental transient absorption at room temperature. AB - The energy transfer rates between chlorophylls in the light harvesting complex CP29 of higher plants at room temperature were calculated ab initio according to the Forster mechanism (Forster T. 1948, Ann. Physik. 2:55-67). Recently, the transition moment orientation of CP29 chlorophylls was determined by differential linear dichroism and absorption spectroscopy of wild-type versus mutant proteins in which single chromophores were missing (Simonetto R., Crimi M., Sandona D., Croce R., Cinque G., Breton J., and Bassi R. 1999. Biochemistry. 38:12974-12983). In this way the Q(y) transition energy and chlorophyll a/b affinity of each binding site was obtained and their characteristics supported by reconstruction of steady-state linear dichroism and absorption spectra at room temperature. In this study, the spectral form of individual chlorophyll a and b ligands within the protein environment was experimentally determined, and their extinction coefficients were also used to evaluate the absolute overlap integral between donors and acceptors employing the Stepanov relation for both the emission spectrum and the Stokes shift. This information was used to calculate the time dependent excitation redistribution among CP29 chlorophylls on solving numerically the Pauli master equation of the complex: transient absorption measurements in the (sub)picosecond time scale were simulated and compared to pump-and-probe experimental data in the Q(y) region on the native CP29 at room temperature upon selective excitation of chlorophylls b at 640 or 650 nm. The kinetic model indicates a bidirectional excitation transfer over all CP29 chlorophylls a species, which is particularly rapid between the pure sites A1-A2 and A4-A5. Chlorophylls b in mixed sites act mostly as energy donors for chlorophylls a, whereas site B5 shows high and bidirectional coupling independent of the pigment hosted. PMID- 11023880 TI - Atomic detail peptide-membrane interactions: molecular dynamics simulation of gramicidin S in a DMPC bilayer. AB - Molecular dynamics simulations have been performed of the sequence-symmetric cyclic decapeptide antibiotic gramicidin S (GS), in interaction with a hydrated dimyristoylphosphatidylcholine (DMPC) bilayer, and the results compared with a "control" simulation of the system in the absence of GS. Following experimental evidence, the GS was initially set in a single antiparallel beta-sheet conformation with two Type II' beta-turns in an amphiphilic interaction with the membrane. This conformation and position remained in the 6.5 ns simulation. Main chain dihedrals are on average approximately 26 degrees from those determined by NMR experiment on GS in dimethylsulfoxide (DMSO) solution. Sequence-symmetric main-chain and side-chain dihedral angle pairs converge to within approximately 5 degrees and approximately 10 degrees, respectively. The area per lipid, lipid tail order parameters, and quadrupole spin-lattice relaxation times of the control simulation are mostly in good agreement with corresponding experiments. The GS has little effect on the membrane dipole potential or water permeability. However, it is found to have a disordering effect (in agreement with experiment) and a fluidifying effect on lipids directly interacting with it, and an ordering effect on those not directly interacting. PMID- 11023881 TI - Thermodynamics and kinetics of a molecular motor ensemble. AB - If, contrary to conventional models of muscle, it is assumed that molecular forces equilibrate among rather than within molecular motors, an equation of state and an expression for energy output can be obtained for a near-equilibrium, coworking ensemble of molecular motors. These equations predict clear, testable relationships between motor structure, motor biochemistry, and ensemble motor function, and we discuss these relationships in the context of various experimental studies. In this model, net work by molecular motors is performed with the relaxation of a near-equilibrium intermediate step in a motor-catalyzed reaction. The free energy available for work is localized to this step, and the rate at which this free energy is transferred to work is accelerated by the free energy of a motor-catalyzed reaction. This thermodynamic model implicitly deals with a motile cell system as a dynamic network (not a rigid lattice) of molecular motors within which the mechanochemistry of one motor influences and is influenced by the mechanochemistry of other motors in the ensemble. PMID- 11023882 TI - Mineralized collagen fibrils: a mechanical model with a staggered arrangement of mineral particles. AB - Both elastic modulus and fracture stress are known to increase with the amount of mineral deposited within collagen fibrils. Current mechanical models of mineralized fibrils, where mineral platelets are arranged in parallel arrays, reproduce the first effect but fail to predict an increase in fracture stress. Here, we propose a model with a staggered array of platelets that is in better agreement with results on molecular packing in collagen fibrils and that accounts for an increase of both elastic modulus and fracture stress with the amount of mineral in the fibril. Finally, we explore the dependence of the mechanical properties within the model, when the degree of mineralization and the thickness of the platelets as well as their distance varies. PMID- 11023883 TI - Lipid demixing and protein-protein interactions in the adsorption of charged proteins on mixed membranes. AB - The adsorption free energy of charged proteins on mixed membranes, containing varying amounts of (oppositely) charged lipids, is calculated based on a mean field free energy expression that accounts explicitly for the ability of the lipids to demix locally, and for lateral interactions between the adsorbed proteins. Minimization of this free energy functional yields the familiar nonlinear Poisson-Boltzmann equation and the boundary condition at the membrane surface that allows for lipid charge rearrangement. These two self-consistent equations are solved simultaneously. The proteins are modeled as uniformly charged spheres and the (bare) membrane as an ideal two-dimensional binary mixture of charged and neutral lipids. Substantial variations in the lipid charge density profiles are found when highly charged proteins adsorb on weakly charged membranes; the lipids, at a certain demixing entropy penalty, adjust their concentration in the vicinity of the adsorbed protein to achieve optimal charge matching. Lateral repulsive interactions between the adsorbed proteins affect the lipid modulation profile and, at high densities, result in substantial lowering of the binding energy. Adsorption isotherms demonstrating the importance of lipid mobility and protein-protein interactions are calculated using an adsorption equation with a coverage-dependent binding constant. Typically, at bulk-surface equilibrium (i.e., when the membrane surface is "saturated" by adsorbed proteins), the membrane charges are "overcompensated" by the protein charges, because only about half of the protein charges (those on the hemispheres facing the membrane) are involved in charge neutralization. Finally, it is argued that the formation of lipid-protein domains may be enhanced by electrostatic adsorption of proteins, but its origin (e.g., elastic deformations associated with lipid demixing) is not purely electrostatic. PMID- 11023884 TI - Diffusion in inhomogeneous media: theory and simulations applied to whole cell photobleach recovery. AB - A continuum description for diffusion in a simple model for an inhomogeneous but isotropic media is derived and implemented numerically. The locally averaged density of diffusible marker is input from experiment to define the sample. Then a single additional parameter, the effective diffusion constant, permits the quantitative simulation of diffusive relaxation from any initial condition. Using this simulation, it is possible to model the recovery of a fluorescently tagged protein in the endoplasmic reticulum (ER) after photobleaching a substantial region of a live cell, and fit an effective diffusion constant which is a property both of the geometry of the ER and the marker. Such quantitative measurements permit inferences about the topology and internal organization of this organelle. PMID- 11023885 TI - Modeling study of exocytosis in neuroendocrine cells: influence of the geometrical parameters. AB - Exocytosis in neuroendocrine cells is a process triggered by Ca(2+). A Monte Carlo simulation of secretion has been developed which, together with the diffusion of calcium, buffered by endogenous and/or exogenously added chelators, also accounts for the dynamics of exocytosis for a pool of readily releasable vesicles. Different distributions of channels and vesicles (random or correlated) are studied. A local study of exocytosis is carried out by obtaining capacitance time courses for the different types of release-ready vesicle pools (correlated or not with Ca(2+) channels). Also, depending upon the kinetic constants for the exocytotic process, we study the levels of local Ca(2+) needed to trigger secretion. Our simulations show that a strong heterogeneity in the calcium concentrations at the different sites of exocytosis is a requirement for reproducing the experimentally observed biphasic response in chromaffin cells in situ (Voets, T., E. Neher, and T. Moser. 1999. Neuron. 23:607-615). Correlated nonuniform distributions of channels and vesicles and the existence of diffusion barriers are shown to quantitatively explain the experimental data on chromaffin cells in situ. The first description requires a deeply heterogeneous distribution, with vesicles attached to the channels or far from them, but never at middle distances. The second description is able to reproduce biphasic release even for uniformly (readily releasable) distributed vesicles. We quantify the degree of inhomogeneity in the distribution of vesicles and how porous the diffusion barriers should be to account for the observed biphasic response. PMID- 11023886 TI - Sequence evolution and the mechanism of protein folding. AB - The impact on protein evolution of the physical laws that govern folding remains obscure. Here, by analyzing in silico-evolved sequences subjected to evolutionary pressure for fast folding, it is shown that: First, a subset of residues in the thermodynamic folding nucleus is mainly responsible for modulating the protein folding rate. Second and most important, the protein topology itself is of paramount importance in determining the location of these residues in the structure. Further stabilization of the interactions in this nucleus leads to fast folding sequences. Third, these nucleation points restrict the sequence space available to the protein during evolution. Correlated mutations between positions around these hot spots arise in a statistically significant manner, and most involve contacting residues. When a similar analysis is carried out on real proteins, qualitatively similar results are obtained. PMID- 11023887 TI - Development and dissipation of Ca(2+) gradients in adrenal chromaffin cells. AB - We used pulsed laser imaging to measure the development and dissipation of Ca(2+) gradients evoked by the activation of voltage-sensitive Ca(2+) channels in adrenal chromaffin cells. Ca(2+) gradients appeared rapidly (<5 ms) upon membrane depolarization and dissipated over several hundred milliseconds after membrane repolarization. Dissipation occurred with an initial fast phase, as the steep gradient near the membrane collapsed, and a slower phase as the remaining shallow gradient dispersed. Inhibition of active Ca(2+) uptake by the endoplasmic reticulum (thapsigargin) and mitochondria (carbonylcyanide p-trifluoro methoxyphenylhydrazone/oligomycin) had no effect on the size of Ca(2+) changes or the rate of gradient dissipation, suggesting that passive endogenous Ca(2+) buffers are responsible for the slow Ca(2+) redistribution. We used a radial diffusion model incorporating Ca(2+) diffusion and binding to intracellular Ca(2+) buffers to simulate Ca(2+) gradients. We included a 3D optical sectioning model, simulating the effects of out-of-focus light, to allow comparison with the measured gradients. Introduction of a high-capacity immobile Ca(2+) buffer, with a buffer capacity on the order of 1000 and appropriate affinity and kinetics, approximated the size of the Ca(2+) increases and rate of dissipation of the measured gradients. Finally, simulations without exogenous buffer suggest that the Ca(2+) signal due to Ca(2+) channel activation is restricted by the endogenous buffer to a space less than 1 microm from the cell membrane. PMID- 11023888 TI - Cytoplasmic molecular delivery with shock waves: importance of impulse. AB - Cell permeabilization using shock waves may be a way of introducing macromolecules and small polar molecules into the cytoplasm, and may have applications in gene therapy and anticancer drug delivery. The pressure profile of a shock wave indicates its energy content, and shock-wave propagation in tissue is associated with cellular displacement, leading to the development of cell deformation. In the present study, three different shock-wave sources were investigated; argon fluoride excimer laser, ruby laser, and shock tube. The duration of the pressure pulse of the shock tube was 100 times longer than the lasers. The uptake of two fluorophores, calcein (molecular weight: 622) and fluorescein isothiocyanate-dextran (molecular weight: 71,600), into HL-60 human promyelocytic leukemia cells was investigated. The intracellular fluorescence was measured by a spectrofluorometer, and the cells were examined by confocal fluorescence microscopy. A single shock wave generated by the shock tube delivered both fluorophores into approximately 50% of the cells (p < 0.01), whereas shock waves from the lasers did not. The cell survival fraction was >0.95. Confocal microscopy showed that, in the case of calcein, there was a uniform fluorescence throughout the cell, whereas, in the case of FITC-dextran, the fluorescence was sometimes in the nucleus and at other times not. We conclude that the impulse of the shock wave (i.e., the pressure integrated over time), rather than the peak pressure, was a dominant factor for causing fluorophore uptake into living cells, and that shock waves might have changed the permeability of the nuclear membrane and transferred molecules directly into the nucleus. PMID- 11023890 TI - Modeling concurrent binding of multiple molecular species in cell adhesion. AB - Cell adhesion provides not only physical linkage but also communication between the cell and its environment. As such, it is important to many cellular functions. Recently, the probability distribution of forming a low number of specific adhesive bonds in a short-duration contact has been described (Chesla et al., Biophys. J. , 1998, 75:1553-1572). This model assumes that binding occurs between a single receptor species and a single ligand species. However, cell adhesion molecules rarely work alone in physiological settings. To account for these in vivo situations, we extended the previous model to include concurrent interactions of multiple receptor-ligand species, introducing the concept of independent binding. Closed-form solutions have been obtained for cases where competition is absent or can be neglected. In two companion papers (Williams et al., Biophys. J., 2000, 79:1858-1866; 2000, 79:1867-1875), the model developed herein has been applied to analyze two sets of experiments designed such that the validity of the theory was also tested. PMID- 11023889 TI - Dynamics of the mitochondrial reticulum in live cells using Fourier imaging correlation spectroscopy and digital video microscopy. AB - We report detailed studies of the dynamics of the mitochondrial reticulum in live cells using two independent experimental techniques: Fourier imaging correlation spectroscopy and digital video fluorescence microscopy. When both methods are used to study the same system, it is possible to directly compare measurements of preaveraged statistical dynamical quantities with their microscopic counterparts. This approach allows the underlying mechanism of the observed rates to be determined. Our results indicate that the dynamics of the reticulum structure is composed of two independent contributions, each important on very different time and length scales. During short time intervals (1-15 sec), local regions of the reticulum primarily undergo constrained thermally activated motion. During long time intervals (>15 sec), local regions of the reticulum undergo long-range "jump" motions that are associated with the action of cytoskeletal filaments. Although the frequency of the jumps depend on the physiological state of the cells, the average jump distance ( approximately 0.8 microm) is unaffected by metabolic activity. During short time intervals, the dynamics appear to be spatially heterogeneous, whereas the cumulative effect of the infrequent jumps leads to the appearance of diffusive motion in the limit of long time intervals. PMID- 11023891 TI - Concurrent binding to multiple ligands: kinetic rates of CD16b for membrane-bound IgG1 and IgG2. AB - CD16b (FcgammaRIIIb) is the most common receptor for the Fc domain of IgG on leukocytes. The binding of Fc receptors to immunoglobulin triggers a wide array of immune responses. In published assays measuring the reaction of CD16b with isotypes of soluble IgG, the affinity for IgG1 was low and that for IgG2 was undetectable. Here we report the first measurement of kinetic rates of CD16b binding to membrane-bound IgG isotypes-a physically distinct and physiologically more relevant presentation-using a recently developed micropipette method. In contrast to the soluble data, we found clearly measurable IgG2 binding, with a forward kinetic rate six-fold lower than that of IgG1 but with an equilibrium affinity only threefold lower. This suggests a nonnegligible role for IgG2 in Fc mediated immune responses, particularly in longer duration contacts. The binding constants were measured from two sets of experiments. Single-isotype experiments were analyzed by an existing model (, Biophys. J. 75:1553-1572). The resulting kinetic rates were used as input to an extended model (, Biophys. J. 79:1850 1857.) to predict the results of mixed-isotype experiments. This design enabled rigorous validation of the concurrent binding model through a test of its predictive ability. PMID- 11023893 TI - Fluctuation of motor charge in the lateral membrane of the cochlear outer hair cell. AB - Functioning of the membrane motor of the outer hair cell is tightly associated with transfer of charge across the membrane. To obtain further insights into the motor mechanism, we examined kinetics of charge transfer across the membrane in two different modes. One is to monitor charge transfer induced by changes in the membrane potential as an excess membrane capacitance. The other is to measure spontaneous flip-flops of charges across the membrane under voltage-clamp conditions as current noise. The noise spectrum of current was inverse Lorentzian, and the capacitance was Lorentzian, as theoretically expected. The characteristic frequency of the capacitance was approximately 10 kHz, and that for current noise was approximately 30 kHz. The difference in the characteristic frequencies seems to reflect the difference in the modes of mechanical movement associated with the two physical quantities. PMID- 11023892 TI - Concurrent and independent binding of Fcgamma receptors IIa and IIIb to surface bound IgG. AB - Fc receptor-antibody interactions are key mechanisms through which antibody effector functions are mediated. Neutrophils coexpress two low-affinity Fcgamma receptors, CD16b (FcgammaRIIIb) and CD32a (FcgammaRIIa), possessing overlapping ligand binding specificities but distinct membrane anchor and signaling capacities. Using K562 cell transfectants as a model, the kinetics of both separate and concurrent binding of CD16b and CD32a to surface-bound IgG ligands were studied. CD16b bound human IgG with 2-3 times higher affinity than did CD32a (A(c)K(a) = 4.1 and 1.6 x 10(-7) microm(4), respectively) and both FcgammaRs had similar reverse kinetic rates (k(r) = 0.5 and 0.4 s(-1), respectively). Because CD16b is expressed on neutrophils at a 4-5 times higher density than CD32a, our results suggest that CD16b plays the dominant role in binding of neutrophils to immobilized IgG. The question of possible cross-regulation of binding affinity between CD16b and CD32a was investigated using our multispecies concurrent binding model (Zhu and Williams, Biophys. J. 79:1850-1857, 2000). Because the model assumes independent binding (no cooperation among different species), the excellent agreement between the model predictions and the experimental data suggests that, when coexpressed on K562 cells, these two FcgammaRs do not interact in a manner that alters the kinetic rates of either molecule. PMID- 11023894 TI - Barrier permeability at cut axonal ends progressively decreases until an ionic seal is formed. AB - After axonal severance, a barrier forms at the cut ends to rapidly restrict bulk inflow and outflow. In severed crayfish axons we used the exclusion of hydrophilic, fluorescent dye molecules of different sizes (0.6-70 kDa) and the temporal decline of ionic injury current to levels in intact axons to determine the time course (0-120 min posttransection) of barrier formation and the posttransection time at which an axolemmal ionic seal had formed, as confirmed by the recovery of resting and action potentials. Confocal images showed that the posttransection time of dye exclusion was inversely related to dye molecular size. A barrier to the smallest dye molecule formed more rapidly (<60 min) than did the barrier to ionic entry (>60 min). These data show that axolemmal sealing lacks abrupt, large changes in barrier permeability that would be expected if a seal were to form suddenly, as previously assumed. Rather, these data suggest that a barrier forms gradually and slowly by restricting the movement of molecules of progressively smaller size amid injury-induced vesicles that accumulate, interact, and form junctional complexes with each other and the axolemma at the cut end. This process eventually culminates in an axolemmal ionic seal, and is not complete until ionic injury current returns to baseline levels measured in an undamaged axon. PMID- 11023895 TI - Adhesive dynamics simulations of sialyl-Lewis(x)/E-selectin-mediated rolling in a cell-free system. AB - Selectin-mediated leukocyte rolling is crucial for the proper function of the immune response. Recently, selectin-mediated rolling was recreated in a cell-free system (Biophysical Journal 71:2902-2907 (1996)); it was shown that sialyl Lewis(x) (sLe(x))-coated microspheres roll over E-selectin-coated surfaces under hydrodynamic flow. The cell-free system removes many confounding cellular features, such as cell deformability and signaling, allowing us to focus on the role of carbohydrate/selectin physical chemistry in mediating rolling. In this paper, we use adhesive dynamics, a computational method that allows us to simulate adhesion, to analyze the experimental data produced in the cell-free system. We simulate the effects of shear rate, ligand density, and number of receptors per particle on rolling velocity and compare them with experimental results obtained with the cell-free system. If we assume the population of particles is homogeneous in receptor density, we predict that particle rolling velocity calculated in simulations is more sensitive to shear rate than found in experiments. Also, the calculated rolling velocity is more sensitive to the number of receptors on the microspheres than to the ligand density on the surface, again in contrast to experiment. We argue that heterogeneity in the distribution of receptors throughout the particle population causes these discrepancies. We improve the agreement between experiment and simulation by calculating the average rolling velocity of a population whose receptors follow a normal distribution, suggesting heterogeneity among particles significantly affects the experimental results. Further comparison between theory and experiment yields an estimate of the reactive compliance of sLe(x)/E-selectin interactions of 0.25 A, close to that reported in the literature for E-selectin and its natural ligand (0.3 A). We also provide an estimate of the value of the intrinsic association rate (between 10(4) and 10(5) s(-1)) for the formation of sLe(x)/E-selectin bonds. PMID- 11023896 TI - Diffusion and deformations of single hydra cells in cellular aggregates. AB - Cell motion within cellular aggregates consists of both random and coherent components. We used confocal microscopy to study the center of mass displacements and deformations of single endodermal Hydra cells in two kinds of cellular aggregates, ectodermal and endodermal. We first carefully characterize the center of mass displacements using standard statistical analysis. In both aggregates, cells perform a persistent random walk, with the diffusion constant smaller in the more cohesive endodermal aggregate. We show that a simple parametric method is able to describe cell deformations and relate them to displacements. These deformations are random, with their amplitude and direction uncorrelated with the center of mass motion. Unlike for an isolated cell on a substrate, the random forces exerted by the surrounding cells predominate over the deformation of the cell itself, causing the displacements of a cell within an aggregate. PMID- 11023897 TI - A direct optimization approach to hidden Markov modeling for single channel kinetics. AB - Hidden Markov modeling (HMM) provides an effective approach for modeling single channel kinetics. Standard HMM is based on Baum's reestimation. As applied to single channel currents, the algorithm has the inability to optimize the rate constants directly. We present here an alternative approach by considering the problem as a general optimization problem. The quasi-Newton method is used for searching the likelihood surface. The analytical derivatives of the likelihood function are derived, thereby maximizing the efficiency of the optimization. Because the rate constants are optimized directly, the approach has advantages such as the allowance for model constraints and the ability to simultaneously fit multiple data sets obtained at different experimental conditions. Numerical examples are presented to illustrate the performance of the algorithm. Comparisons with Baum's reestimation suggest that the approach has a superior convergence speed when the likelihood surface is poorly defined due to, for example, a low signal-to-noise ratio or the aggregation of multiple states having identical conductances. PMID- 11023898 TI - Hidden Markov modeling for single channel kinetics with filtering and correlated noise. AB - Hidden Markov modeling (HMM) can be applied to extract single channel kinetics at signal-to-noise ratios that are too low for conventional analysis. There are two general HMM approaches: traditional Baum's reestimation and direct optimization. The optimization approach has the advantage that it optimizes the rate constants directly. This allows setting constraints on the rate constants, fitting multiple data sets across different experimental conditions, and handling nonstationary channels where the starting probability of the channel depends on the unknown kinetics. We present here an extension of this approach that addresses the additional issues of low-pass filtering and correlated noise. The filtering is modeled using a finite impulse response (FIR) filter applied to the underlying signal, and the noise correlation is accounted for using an autoregressive (AR) process. In addition to correlated background noise, the algorithm allows for excess open channel noise that can be white or correlated. To maximize the efficiency of the algorithm, we derive the analytical derivatives of the likelihood function with respect to all unknown model parameters. The search of the likelihood space is performed using a variable metric method. Extension of the algorithm to data containing multiple channels is described. Examples are presented that demonstrate the applicability and effectiveness of the algorithm. Practical issues such as the selection of appropriate noise AR orders are also discussed through examples. PMID- 11023899 TI - Na(+)-Ca(2+)-K(+) currents measured in insect cells transfected with the retinal cone or rod Na(+)-Ca(2+)-K(+) exchanger cDNA. AB - The recently cloned retinal cone Na(+)-Ca(2+)-K(+) exchanger (NCKX) was expressed in cultured insect cells, and whole-cell patch clamp was used to measure transmembrane currents generated by this transcript and compare them with currents generated by retinal rod NCKX or by a deletion mutant rod NCKX from which the two large hydrophilic loops were removed. We have characterized the ionic currents generated by both the forward (Ca(2+) extrusion) and reverse (Ca(2+) influx) modes of all three NCKX proteins. Reverse NCKX exchange generated outward current that required the simultaneous presence of both external Ca(2+) and external K(+). Forward NCKX exchange carried inward current with Na(+), but not with Li(+) in the bath solution. The cation dependencies of the three NCKX tested (external K(+), external Na(+), internal Ca(2+)) were very similar to each other and to those reported previously for the in situ rod NCKX. These findings provide the first electrophysiological characterization of cone NCKX and the first electrophysiological characterization of potassium-dependent Na(+)-Ca(+) exchangers in heterologous systems. Our results demonstrate the feasibility of combining heterologous expression and biophysical measurements for detailed NCKX structure/function studies. PMID- 11023900 TI - Properties of gap junction channels formed by Cx46 alone and in combination with Cx50. AB - Gap junctions formed of connexin46 (Cx46) and connexin50 (Cx50) in lens fiber cells are crucial for maintaining lens transparency. We determined the functional properties of homotypic Cx46, heterotypic Cx46/Cx50, and heteromeric Cx46/Cx50 channels in a communication-deficient neuroblastoma (N2A) cell line, using dual whole-cell recordings. N2A cultures were stably and/or transiently transfected with Cx46, Cx50, and green fluorescent protein (EGFP). The macroscopic voltage sensitivity of homotypic Cx46 conformed to the two-state model (Boltzmann parameters: G(min) = 0.11, V(0) = +/- 48.1 mV, gating charge = 2). Cx46 single channels showed a main-state conductance of 140 +/- 8 pS and multiple subconductance states ranging from < or =10 pS to 60 pS. Conservation of homotypic properties in heterotypic Cx46/Cx50 cell pairs allowed the determination of a positive relative gating polarity for the dominant gating mechanisms in Cx46 and Cx50. Observed gating properties were consistent with a second gating mechanism in Cx46 connexons. Moreover, rectification was observed in heterotypic cell pairs. Some cell pairs in cultures simultaneously transfected with Cx46 and Cx50 exhibited junctional properties not observed in other preparations, suggesting the formation of heteromeric channels. We conclude that different combinations of Cx46 and Cx50 within gap junction channels lead to unique biophysical properties. PMID- 11023901 TI - Interaction of the noncovalent molecular adapter, beta-cyclodextrin, with the staphylococcal alpha-hemolysin pore. AB - Cyclodextrins act as noncovalent molecular adapters when lodged in the lumen of the alpha-hemolysin (alphaHL) pore. The adapters act as binding sites for channel blockers, thereby offering a basis for the detection of a variety of organic molecules with alphaHL as a biosensor element. To further such studies, it is important to find conditions under which the dwell time of cyclodextrins in the lumen of the pore is extended. Here, we use single-channel recording to explore the pH- and voltage-dependence of the interaction of beta-cyclodextrin (betaCD) with alphaHL. betaCD can access its binding site only from the trans entrance of pores inserted from the cis side of a bilayer. Analysis of the binding kinetics shows that there is a single binding site for betaCD, with an apparent equilibrium dissociation constant that varies by >100-fold under the conditions explored. The dissociation rate constant for the neutral betaCD molecule varies with pH and voltage, a result that is incompatible with two states of the alphaHL pore, one of high and the other of low affinity. Rather, the data suggest that the actual equilibrium dissociation constant for the alphaHL. betaCD complex varies continuously with the transmembrane potential. PMID- 11023902 TI - Binding and selectivity in L-type calcium channels: a mean spherical approximation. AB - L-type calcium channels are Ca(2+) binding proteins of great biological importance. They generate an essential intracellular signal of living cells by allowing Ca(2+) ions to move across the lipid membrane into the cell, thereby selecting an ion that is in low extracellular abundance. Their mechanism of selection involves four carboxylate groups, containing eight oxygen ions, that belong to the side chains of the "EEEE" locus of the channel protein, a setting similar to that found in many Ca(2+)-chelating molecules. This study examines the hypothesis that selectivity in this locus is determined by mutual electrostatic screening and volume exclusion between ions and carboxylate oxygens of finite diameters. In this model, the eight half-charged oxygens of the tethered carboxylate groups of the protein are confined to a subvolume of the pore (the "filter"), but interact spontaneously with their mobile counterions as ions interact in concentrated bulk solutions. The mean spherical approximation (MSA) is used to predict ion-specific excess chemical potentials in the filter and baths. The theory is calibrated using a single experimental observation, concerning the apparent dissociation constant of Ca(2+) in the presence of a physiological concentration of NaCl. When ions are assigned their independently known crystal diameters and the carboxylate oxygens are constrained, e.g., to a volume of 0.375 nm(3) in an environment with an effective dielectric coefficient of 63.5, the hypothesized selectivity filter produces the shape of the calcium binding curves observed in experiment, and it predicts Ba(2+)/Ca(2+) and Na(+)/Li(+) competition, and Cl(-) exclusion as observed. The selectivities for Na(+), Ca(2+), Ba(2+), other alkali metal ions, and Cl(-) thus can be predicted by volume exclusion and electrostatic screening alone. Spontaneous coordination of ions and carboxylates can produce a wide range of Ca(2+) selectivities, depending on the volume density of carboxylate groups and the permittivity in the locus. A specific three-dimensional structure of atoms at the binding site is not needed to explain Ca(2+) selectivity. PMID- 11023903 TI - Characterization of single-cell electroporation by using patch-clamp and fluorescence microscopy. AB - Electroporation of single NG108-15 cells with carbon-fiber microelectrodes was characterized by patch-clamp recordings and fluorescence microscopy. To minimize adverse capacitive charging effects, the patch-clamp pipette was sealed on the cell at a 90(o) angle with respect to the microelectrodes where the applied potential reaches a minimum. From transmembrane current responses, we determined the electric field strengths necessary for ion-permeable pore formation and investigated the kinetics of pore opening and closing as well as pore open times. From both patch-clamp and fluorescence microscopy experiments, the threshold transmembrane potentials for dielectric breakdown of NG108-15 cells, using 1-ms rectangular waveform pulses, was approximately 250 mV. The electroporation pulse preceded pore formation, and analyte entry into the cells was dictated by concentration, and membrane resting potential driving forces. By stepwise moving a cell out of the focused field while measuring the transmembrane current response during a supramaximal pulse, we show that cells at a distance of approximately 30 microm from the focused field were not permeabilized. PMID- 11023905 TI - Differential effects of surfactant protein A on regional organization of phospholipid monolayers containing surfactant protein B or C. AB - Epifluorescence microscopy combined with a surface balance was used to study monolayers of dipalmitoylphosphatidylcholine (DPPC)/egg phosphatidylglycerol (PG) (8:2, mol/mol) plus 17 wt % SP-B or SP-C spread on subphases containing SP-A in the presence or absence of 5 mM Ca(2+). Independently of the presence of Ca(2+) in the subphase, SP-A at a bulk concentration of 0.68 microg/ml adsorbed into the spread monolayers and caused an increase in the molecular areas in the films. Films of DPPC/PG formed on SP-A solutions showed a pressure-dependent coexistence of liquid-condensed (LC) and liquid-expanded (LE) phases. Apart from these surface phases, a probe-excluding phase, likely enriched in SP-A, was seen in the films between 7 mN/m < or = pi < or = 20 mN/m. In monolayers of SP-B/(DPPC/PG) spread on SP-A, regardless of the presence of calcium ions, large clusters of a probe-excluding phase, different from probe-excluding lipid LC phase, appeared and segregated from the LE phase at near-zero surface pressures and coexisted with the conventional LE and LC phases up to approximately 35 mN/m. Varying the levels of either SP-A or SP-B in films of SP-B/SP-A/(DPPC/PG) revealed that the formation of the probe-excluding clusters distinctive for the quaternary films was influenced by the two proteins. Concanavalin A in the subphase could not replace SP-A in its ability to modulate the textures of films of SP-B/(DPPC/PG). In films of SP-C/SP-A/(DPPC/PG), in the absence of calcium, regions consisting of a probe-excluding phase, likely enriched in SP-A, were detected at surface pressures between 2 mN/m and 20 mN/m in addition to the lipid LE and LC phases. Ca(2+) in the subphase appeared to disperse this phase into tiny probe-excluding particles, likely comprising Ca(2+)-aggregated SP-A. Despite their strikingly different morphologies, the films of DPPC/PG that contained combinations of SP B/SP-A or SP-C/SP-A displayed similar distributions of LC and LE phases with LC regions occupying a maximum of 20% of the total monolayer area. Combining SP-A and SP-B reorganized the morphology of monolayers composed of DPPC and PG in a Ca(2+)-independent manner that led to the formation of a separate potentially protein-rich phase in the films. PMID- 11023904 TI - Crystallization of antimicrobial pores in membranes: magainin and protegrin. AB - Membrane pores spontaneously formed by antimicrobial peptides in membranes were crystallized for the first time by manipulating the sample hydration and temperature. Neutron diffraction shows that magainins and protegrins form stable pores in fully hydrated fluid membranes. At lower hydration levels or low temperature, the membrane multilayers crystallize. In one crystalline phase, the pores in each bilayer arrange in a regular hexagonal array and the bilayers are stacked into a hexagonal ABC lattice, corresponding to the cubic close-packed structure of spheres. In another crystalline phase, the bilayers are modulated into the rippled multilamellae, corresponding to a 2D monoclinic lattice. The phase diagrams are described. Crystallization of the membrane pores provides possibilities for diffraction studies that might provide useful information on the pore structures. PMID- 11023906 TI - The EGF receptor transmembrane domain: peptide-peptide interactions in fluid bilayer membranes. AB - A peptide containing the transmembrane domain of the human EGF receptor was studied in fluid lipid bilayers for insight into receptor tyrosine kinase lateral associations in cell membranes. The peptide comprised the 23-amino acid hydrophobic segment thought to span the membrane (Ile(622) to Met(644) of the EGF receptor), plus the first 10 amino acids of the receptor's cytoplasmic domain (Arg(645) to Thr(654)). Probes for solid-state NMR spectroscopy were incorporated by deuteration of the methyl side chains of alanine at positions 623 and 637. (2)H-NMR spectra were recorded from 25 to 65 degrees C in membranes composed of 1 palmitoyl-2-oleoyl phosphatidylcholine, with and without 33% cholesterol, and relaxation times were measured. Peptide concentration ranged from 0. 5 to 10 mol %. The peptide behaved as predominant monomers undergoing rapid symmetric rotational diffusion; however, there was evidence of reversible side-to-side interaction among the hydrophobic transmembrane domains, particularly at physiological temperatures and in the presence of natural concentrations of cholesterol. The results of these experiments in fluid membranes are consistent with the existence of lipid-protein interactions that would predispose to receptor microdomain formation in membranes of higher animal cells. PMID- 11023907 TI - Miscibility critical pressures in monolayers of ternary lipid mixtures. AB - When phospholipids are mixed with cholesterol in a monolayer at an air-water interface, coexisting 2-dimensional liquid phases can be observed if the surface pressure, pi, is lower than the miscibility critical pressure, pi(c). Ternary mixtures of two phospholipid species with dihydrocholesterol have been reported to have critical pressures that are linearly proportional to the relative composition of the phospholipids. However, we report here that, if the acyl chains of the two phospholipids differ significantly in length or unsaturation, the behavior is markedly different. In this case, the critical pressure of the ternary mixture can be remarkably high, exceeding the critical pressures of the corresponding binary mixtures. High critical pressures are also seen in binary mixtures of phospholipid and dihydrocholesterol when the two acyl chains of the phospholipid differ sufficiently in length. Using regular solution theory, we interpret the elevated critical pressures of these mixtures as an attractive interaction between the phospholipid components. PMID- 11023908 TI - Calorimetric and spectroscopic studies of the thermotropic phase behavior of lipid bilayer model membranes composed of a homologous series of linear saturated phosphatidylserines. AB - The thermotropic phase behavior of lipid bilayer model membranes composed of the even-numbered, N-saturated 1,2-diacyl phosphatidylserines was studied by differential scanning calorimetry and by Fourier-transform infrared and (31)P nuclear magnetic resonance spectroscopy. At pH 7.0, 0.1 M NaCl and in the absence of divalent cations, aqueous dispersions of these lipids, which have not been incubated at low temperature, exhibit a single calorimetrically detectable phase transition that is fully reversible, highly cooperative, and relatively energetic, and the transition temperatures and enthalpies increase progressively with increases in hydrocarbon chain length. Our spectroscopic observations confirm that this thermal event is a lamellar gel (L(beta))-to-lamellar liquid crystalline (L(alpha)) phase transition. However, after low temperature incubation, the L(beta)/L(alpha) phase transition of dilauroyl phosphatidylserine is replaced by a higher temperature, more enthalpic, and less cooperative phase transition, and an additional lower temperature, less enthalpic, and less cooperative phase transition appears in the longer chain phosphatidylserines. Our spectroscopic results indicate that this change in thermotropic phase behavior when incubated at low temperatures results from the conversion of the L(beta) phase to a highly ordered lamellar crystalline (L(c)) phase. Upon heating, the L(c) phase of dilauroyl phosphatidylserine converts directly to the L(alpha) phase at a temperature slightly higher than that of its original L(beta)/L(alpha) phase transition. Calorimetrically, this process is manifested by a less cooperative but considerably more energetic, higher-temperature phase transition, which replaces the weaker L(beta)/L(alpha) phase transition alluded to above. However, with the longer chain compounds, the L(c) phase first converts to the L(beta) phase at temperatures some 10-25 degrees C below that at which the L(beta) phase converts to the L(alpha) phase. Our results also suggest that shorter chain homologues form L(c) phases that are structurally related to, but more ordered than, those formed by the longer chain homologues, but that these L(c) phases are less ordered than those formed by other phospholipids. These studies also suggest that polar/apolar interfaces of the phosphatidylserine bilayers are more hydrated than those of other glycerolipid bilayers, possibly because of interactions between the polar headgroup and carbonyl groups of the fatty acyl chains. PMID- 11023909 TI - Differential scanning calorimetric and Fourier transform infrared spectroscopic studies of the effects of cholesterol on the thermotropic phase behavior and organization of a homologous series of linear saturated phosphatidylserine bilayer membranes. AB - We have examined the effects of cholesterol on the thermotropic phase behavior and organization of aqueous dispersions of a homologous series of linear disaturated phosphatidylserines by high-sensitivity differential scanning calorimetry and Fourier transform infrared spectroscopy. We find that the incorporation of increasing quantities of cholesterol progressively reduces the temperature, enthalpy, and cooperativity of the gel-to-liquid-crystalline phase transition of the host phosphatidylserine bilayer, such that a cooperative chain melting phase transition is completely or almost completely abolished at 50 mol % cholesterol, in contrast to the results of previous studies. We are also unable to detect the presence of a separate anhydrous cholesterol or cholesterol monohydrate phase in our binary mixtures, again in contrast to previous reports. We further show that the magnitude of the reduction in the phase transition temperature induced by cholesterol addition is independent of the hydrocarbon chain length of the phosphatidylserine studied. This result contrasts with our previous results with phosphatidylcholine bilayers, where we found that cholesterol increases or decreases the phase transition temperature in a chain length-dependent manner (1993. Biochemistry, 32:516-522), but is in agreement with our previous results for phosphatidylethanolamine bilayers, where no hydrocarbon chain length-dependent effects were observed (1999. Biochim. Biophys. Acta, 1416:119-234). However, the reduction in the phase transition temperature by cholesterol is of greater magnitude in phosphatidylethanolamine as compared to phosphatidylserine bilayers. We also show that the addition of cholesterol facilitates the formation of the lamellar crystalline phase in phosphatidylserine bilayers, as it does in phosphatidylethanolamine bilayers, whereas the formation of such phases in phosphatidylcholine bilayers is inhibited by the presence of cholesterol. We ascribe the limited miscibility of cholesterol in phosphatidylserine bilayers reported previously to a fractional crystallization of the cholesterol and phospholipid phases during the removal of organic solvent from the binary mixture before the hydration of the sample. In general, the results of our studies to date indicate that the magnitude of the effect of cholesterol on the thermotropic phase behavior of the host phospholipid bilayer, and its miscibility in phospholipid dispersions generally, depend on the strength of the attractive interactions between the polar headgroups and the hydrocarbon chains of the phospholipid molecule, and not on the charge of the polar headgroups per se. PMID- 11023910 TI - Selectivity in lipid binding to the bacterial outer membrane protein OmpF. AB - The outer membrane porin OmpF from Escherichia coli has been reconstituted into lipid bilayers of defined composition, and fluorescence spectroscopy is used to characterize its interaction with the surrounding lipid. OmpF is a trimer within the membrane. It contains two Trp residues per monomer, Trp(214) at the lipid protein interface and Trp(61) at the trimer interface. The fluorescence of Trp 214 in the mutant W61F is quenched by dibromostearoylphosphatidylcholine (di(Br(2)C18:0)PC), whereas the fluorescence of Trp(61) in the mutant W214F is not quenched by di(Br(2)C18:0)PC when fluorescence is excited directly through the Trp rather than through the Tyr residues. Measurements of relative fluorescence quenching for OmpF reconstituted into mixtures of lipid X and di(Br(2)C18:0)PC have been analyzed to give the binding constant of lipid X for OmpF, relative to that for dioleoylphosphatidylcholine (di(C18:1)PC). The phosphatidylcholine showing the strongest binding to OmpF is dimyristoyloleoylphosphatidylcholine (di(C14:1)PC) with binding constants decreasing with either increasing or decreasing fatty acyl chain length. Comparison with various theories for hydrophobic matching between lipids and proteins suggests that in the chain length range from C14 to C20, hydrophobic matching is achieved largely by distortion of the lipid bilayer around the OmpF, whereas for chains longer than C20, distortion of both the lipid bilayer and of the protein is required to achieve hydrophobic matching. Phosphatidylcholine and phosphatidylethanolamine bind with equal affinity to OmpF, but the affinity for phosphatidylglycerol is about half that for phosphatidylcholine. PMID- 11023911 TI - Polar angle as a determinant of amphipathic alpha-helix-lipid interactions: a model peptide study. AB - Various physicochemical properties play important roles in the membrane activities of amphipathic antimicrobial peptides. To examine the effects of the polar angle, two model peptides, thetap100 and thetap180, with polar angles of 100 degrees and 180 degrees, respectively, were designed, and their interactions with membranes were investigated in detail. These peptides have almost identical physicochemical properties except for polar angle. Like naturally occurring peptides, these peptides selectively bind to acidic membranes, assuming amphipathic alpha-helices, and formed peptide-lipid supramolecular complex pores accompanied by lipid flip-flop and peptide translocation. Despite its somewhat lower membrane affinity, thetap100 exhibited higher membrane permeabilization activity, a greater flip-flop rate, as well as more antimicrobial activity due to a higher pore formation rate compared with thetap180. Consistent with these results, the peptide translocation rate of thetap100 was higher. Furthermore, the number of peptides constituting thetap100 pores was less than that of thetap180, and thetap100 pores involved more lipid molecules, as reflected by its cation selectivity. The polar angle was found to be an important parameter determining peptide-lipid interactions. PMID- 11023913 TI - Actinomycin D binds strongly to d(CGACGACG) and d(CGTCGTCG). AB - Earlier calorimetric studies had indicated that despite the absence of a GpC sequence, the self-complementary octamer d(CGTCGACG) binds strongly to actinomycin D (ACTD) with high cooperativity and a 2:1 drug/duplex ratio. A subsequent optical spectral study with related oligomers led us to suggest that ACTD may likely stack at the G. C basepairs of the duplex termini. New findings are reported herein to indicate that despite the lack of complete self complementarity, oligomers of d(CGXCGXCG) [X = A or T] motif exhibit unusually strong ACTD affinities with binding constants of roughly 2 x 10(7) M(-1) and binding densities of 1 drug molecule per strand. The ACTD binding affinity for the corresponding heteroduplex obtained by annealing these two oligomers is, however, considerably reduced. Although spectroscopic results with related oligomers obtained by removing, replacing, or appending bases at the termini appear to be consistent with the end-stacking model, capillary electrophoretic (CE) evidence provides additional insights into the binding mode. CE experiments with the self-complementary oligomers d(CGAGCTCG) and d(CGTCGACG) revealed contrasting migration patterns in the presence of ACTD, with mobility retardation and acceleration exhibited by the GpC- and non-GpC-containing octamers, respectively, whereas the X/X-mismatched d(CGXCGXCG) experienced retardation. These results, along with those of related oligomers, suggest that ACTD may in fact stack at the duplex stem end of a monomeric hairpin or at the 3'-end of dG as a single strand. The seemingly cooperative ACTD binding and the curved Scatchard plot for the self-complementary d(CGTCGACG) may thus be attributed to the drug-induced duplex denaturation resulting from strong binding to single strands of d(CGXCGYCG) motif. Detailed structural information on the ACTD-DNA complexes, however, must await further NMR investigations. PMID- 11023912 TI - Protein diffusion in living skeletal muscle fibers: dependence on protein size, fiber type, and contraction. AB - Sarcoplasmic protein diffusion was studied under different conditions, using microinjection in combination with microspectrophotometry. Six globular proteins with molecular masses between 12 and 3700 kDa, with diameters from 3 to 30 nm, were used for the experiments. Proteins were injected into single, intact skeletal muscle fibers taken from either soleus or extensor digitorum longus (edl) muscle of adult rats. No correlation was found between sarcomere spacing and the sarcoplasmic diffusion coefficient (D) for all proteins studied. D of the smaller proteins cytochrome c (diameter 3.1 nm), myoglobin (diameter 3.5 nm), and hemoglobin (diameter 5.5 nm) amounted to only approximately 1/10 of their value in water and was not increased by auxotonic fiber contractions. D for cytochrome c and myoglobin was significantly higher in fibers from edl (mainly type II fibers) compared to fibers from soleus (mainly type I fibers). Measurements of D for myoglobin at 37 degrees C in addition to 22 degrees C led to a Q(10) of 1.46 for this temperature range. For the larger proteins catalase (diameter 10.5 nm) and ferritin (diameter 12.2 nm), a decrease in D to approximately 1/20 and approximately 1/50 of that in water was observed, whereas no diffusive flux at all of earthworm hemoglobin (diameter 30 nm) along the fiber axis could be detected. We conclude that 1) sarcoplasmic protein diffusion is strongly impaired by the presence of the myofilamental lattice, which also gives rise to differences in diffusivity between different fiber types; 2) contractions do not cause significant convection in sarcoplasm and do not lead to increased diffusional transport; and 3) in addition to the steric hindrance that slows down the diffusion of smaller proteins, diffusion of large proteins is further hindered when their dimensions approach the interfilament distances. This molecular sieve property progressively reduces intracellular diffusion of proteins when the molecular diameter increases to more than approximately 10 nm. PMID- 11023914 TI - Exciton dynamics in the chlorosomal antennae of the green bacteria Chloroflexus aurantiacus and Chlorobium tepidum. AB - The energy transfer processes in isolated chlorosomes from green bacteria Chlorobium tepidum and Chloroflexus aurantiacus have been studied at low temperatures (1.27 K) by two-pulse photon echo and one-color transient absorption techniques with approximately 100 fs resolution. The decay of the coherence in both types of chlorosomes is characterized by four different dephasing times stretching from approximately 100 fs up to 300 ps. The fastest component reflects dephasing that is due to interaction of bacteriochlorophylls with the phonon bath, whereas the other components correspond to dephasing due to different energy transfer processes such as distribution of excitation along the rod-like aggregates, energy exchange between different rods in the chlorosome, and energy transfer to the base plate. As a basis for the interpretation of the excitation dephasing and energy transfer pathways, a superlattice-like structural model is proposed based on recent experimental data and computer modeling of the Bchl c aggregates (1994. Photosynth. Res. 41:225-233.) This model predicts a fine structure of the Q(y) absorption band that is fully supported by the present photon echo data. PMID- 11023915 TI - Membrane photopotential generation by interfacial differences in the turnover of a photodynamic reaction. AB - The adsorption of a membrane-impermeable photosensitizer to only one membrane leaflet is found to trigger a localized photodynamic reaction; i.e., the amount of carbonyl cyanide m-chlorophenylhydrazone (CCCP) molecules damaged in the leaflet facing the photosensitizer is roughly identical to the total amount of CCCP inactivated. Whereas the latter quantity is assessed from the drop in membrane conductivity G, the former is evaluated from the photopotential phi that is proportional to the interfacial concentration difference of the uncoupler. Localized photodestruction is encountered by CCCP diffusion to the site of photodamage. A simple model that accounts for both photoinhibition and diffusion predicts the dependence of the photopotential on light intensity, buffer capacity, and pH of the medium. It is concluded that only a limited amount of the reactive oxygen species responsible for CCCP photodamage diffuses across the membrane. If the concentration of reactive oxygen species is decreased by addition of NaN(3) or by substituting aqueous oxygen for argon, phi is inhibited. If, in contrast, their life time is increased by substitution of H(2)O for D(2)O, phi increases. PMID- 11023916 TI - Early intermediates in the photocycle of the Glu46Gln mutant of photoactive yellow protein: femtosecond spectroscopy. AB - Transient absorption spectroscopy in the time range from -1 ps to 4 ns, and over the wavelength range from 420 to 550 nm, was applied to the Glu46Gln mutant of the photoactive yellow protein (PYP) from Ectothiorhodospira halophila. This has allowed us to elucidate the kinetic constants of excited state formation and decay and photochemical product formation, and the spectral characteristics of stimulated emission and the early photocycle intermediates. Both the quantum efficiency ( approximately 0.5) and the rate constants for excited state decay and the formation of the initial photochemical intermediate (I(0)) were found to be quite similar to those obtained for wild-type PYP. In contrast, the rate constants for the formation of the subsequent photocycle intermediates (I(0)(double dagger) and I(1)), as well as for I(2) and for ground state regeneration as determined in earlier studies, were found to be from 3- to 30 fold larger. The structural implications of these results are discussed. PMID- 11023917 TI - pH- and temperature-dependence of functional modulation in metalloproteinases. A comparison between neutrophil collagenase and gelatinases A and B. AB - Metalloproteases are metalloenzymes secreted in the extracellular fluid and involved in inflammatory pathologies or events, such as extracellular degradation. A Zn(2+) metal, present in the active site, is involved in the catalytic mechanism, and it is generally coordinated with histidyl and/or cysteinyl residues of the protein moiety. In this study we have investigated the effect of both pH (between pH 4.8 and 9.0) and temperature (between 15 degrees C and 37 degrees C) on the enzymatic functional properties of the neutrophil interstitial collagenase (MMP-8), gelatinases A (MMP-2) and B (MMP-9), using the same synthetic substrate, namely MCA-Pro-Leu-Gly approximately Leu-DPA-Ala-Arg NH(2). A global analysis of the observed proton-linked behavior for k(cat)/K(m), k(cat), and K(m) indicates that in order to have a fully consistent description of the enzymatic action of these metalloproteases we have to imply at least three protonating groups, with differing features for the three enzymes investigated, which are involved in the modulation of substrate interaction and catalysis by the enzyme. This is the first investigation of this type on recombinant collagenases and gelatinases of human origin. The functional behavior, although qualitatively similar, displays significant differences with respect to what was previously observed for stromelysin and porcine collagenase and gelatinase (Stack, M. S., and R. D. Gray. 1990. Arch. Biochem. Biophys. 281:257-263; Harrison, R. K., B. Chang, L. Niedzwiecki, and R. L. Stein. 1992. Biochemistry. 31:10757-10762). The functional characterization of these enzymes can have some relevant physiological significance, since it may be related to the marked changes in the environmental pH that collagenase and gelatinases may experience in vivo, moving from the intracellular environment to the extracellular matrix. PMID- 11023918 TI - The unfolding/denaturation of immunogammaglobulin of isotype 2b and its F(ab) and F(c) fragments. AB - The unfolding and further denaturation of IgG and its F(ab) and F(c) fragments were studied both on a macroscopic and molecular level, using differential scanning calorimetry and circular dichroism spectroscopy, respectively. It was shown that the structural integrity of the F(ab) and F(c) units was retained after fragmentation of the IgG. The F(ab) fragment denatured at approximately 61 degrees C and the F(c) fragment at 71 degrees C. The structural transitions observed in the whole IgG is the sum effect of those determined for the isolated F(ab) and F(c) fragments. PMID- 11023920 TI - The noise of membrane capacitance measurements in the whole-cell recording configuration. AB - High-resolution measurement of membrane capacitance in the whole-cell-recording configuration can be used to detect small changes in membrane surface area that accompany exocytosis and endocytosis. We have investigated the noise of membrane capacitance measurements to determine the fundamental limits of resolution in actual cells in the whole-cell mode. Two previously overlooked sources of noise are particularly evident at low frequencies. The first noise source is accompanied by a correlation between capacitance estimates, whereas the second noise source is due to "1/f-like" current noise. An analytic expression that summarizes the noise from thermal and 1/f sources is derived, which agrees with experimental measurements from actual cells over a large frequency range. Our results demonstrate that the optimal frequencies for capacitance measurements are higher than previously believed. Finally, we demonstrate that the capacitance noise at high frequencies can be reduced by compensating for the voltage drop of the sine wave across the series resistance. PMID- 11023919 TI - Radiation inactivation of ribonucleotide reductase, an enzyme with a stable free radical. AB - Herpes simplex virus ribonucleotide reductase (RR) is a tetrameric enzyme composed of two homodimers of large R1 and small R2 subunits with a tyrosyl free radical located on the small subunit. Irradiation of the holoenzyme yielded simple exponential decay curves and an estimated functional target size of 315 kDa. Western blot analysis of irradiated holoenzyme R1 and R2 yielded target sizes of 281 kDa and 57 kDa (approximately twice their expected size). Irradiation of free R1 and analysis by all methods yielded a single exponential decay with target sizes ranging from 128-153 kDa. For free R2, quantitation by enzyme activity and Western blot analyses yielded simple inactivation curves but considerably different target sizes of 223 kDa and 19 kDa, respectively; competition for radioligand binding in irradiated R2 subunits yielded two species, one with a target size of approximately 210 kDa and the other of approximately 20 kDa. These results are consistent with a model in which there is radiation energy transfer between the two monomers of both R1 and R2 only in the holoenzyme, a radiation-induced loss of free radical only in the isolated R2, and an alteration of the tertiary structure of R2. PMID- 11023921 TI - Cation-binding sites of subtilisin Carlsberg probed with Eu(III) luminescence. AB - Two Ca(2+)-binding sites of subtilisin Carlsberg are studied by monitoring static and time-resolved luminescence of selectively substituted Eu(3+) at each site, and they are found to be characteristically quite different from each other. Compared with the coordination sphere of free Eu(3+), two sites are very similar to each other, so that both have a well-defined binding structure with low coordination symmetry. However, compared with the weak site, the strong site is relatively more polar, more symmetrical, and more easily accessible. Furthermore, despite the absence of water reported in the x-ray crystal structure (, Eur. J. Biochem. 166:673-692), one water molecule is found to exist in the coordination sphere of the strong site in aqueous solution. Thus it is suggested that in solution the Ca(2+) bound in the strong site forms an additional coordination bond to a solvent or substrate molecule. PMID- 11023922 TI - Analysis of sedimentation equilibrium distributions reflecting nonideal macromolecular associations. AB - A rigorous statistical-mechanical approach is adopted to derive general quantitative expressions that allow for the effects of thermodynamic nonideality in equilibrium measurements reflecting interaction between dissimilar macromolecular reactants. An analytical procedure based on these expressions is then formulated for obtaining global estimates of equilibrium constants and the corresponding reference thermodynamic activities of the free reactants in each of several sedimentation equilibrium experiments. The method is demonstrated by application to results from an ultracentrifugal study of an electrostatic interaction between ovalbumin and cytochrome c (Winzor, D. J., M. P. Jacobsen, and P. R. Wills. 1998. Biochemistry. 37:2226-2233). It is demonstrated that reliable estimates of relevant thermodynamic parameters are extracted from the data through statistical analysis by means of a simple nonlinear fitting procedure. PMID- 11023923 TI - Tracking single proteins within cells. AB - We present experiments in which single proteins were imaged and tracked within mammalian cells. Single proteins of R-phycoerythrin (RPE) were imaged by epifluorescence microscopy in the nucleoplasm and cytoplasm at 71 frames/s. We acquired two-dimensional trajectories of proteins (corresponding to the projection of three-dimensional trajectories onto the plane of focus) for an average of 17 frames in the cytoplasm and 16 frames in the nucleus. Diffusion constants were determined from linear fits to the mean square displacement and from the mean displacement squared per frame. We find that the distribution of diffusion constants for RPE within cells is broader than the distributions obtained from RPE in a glycerol solution, from a Monte Carlo simulation, and from the theoretical distribution for simple diffusion. This suggests that on the time scales of our measurements, the motion of single RPE proteins in the cytoplasm and nucleoplasm cannot be modeled by simple diffusion with a unique diffusion constant. Our results demonstrate that it is possible to follow the motion of single proteins within cells and that the technique of single molecule tracking can be used to probe the dynamics of intracellular macromolecules. PMID- 11023924 TI - The use of pHluorins for optical measurements of presynaptic activity. AB - Genetically encoded reporters for optical measurements of presynaptic activity hold significant promise for measurements of neurotransmission within intact or semi-intact neuronal networks. We have characterized pH-sensitive green fluorescent protein-based sensors (pHluorins) of synaptic vesicle cycling at nerve terminals. pHluorins have a pK approximately 7.1, which make them ideal for tracking synaptic vesicle lumen pH upon cycling through the plasma membrane during action potentials. A theoretical analysis of the expected signals using this approach and guidelines for future reporter development are provided. PMID- 11023925 TI - Secondary prevention antibiotic treatment trials for coronary artery disease. PMID- 11023926 TI - Functional effect of the C242T polymorphism in the NAD(P)H oxidase p22phox gene on vascular superoxide production in atherosclerosis. AB - BACKGROUND: Increased superoxide anion production increases oxidative stress and reduces nitric oxide bioactivity in vascular disease states. NAD(P)H oxidase is an important source of superoxide in human blood vessels, and some studies suggest a possible association between polymorphisms in the NAD(P)H oxidase CYBA gene and atherosclerosis; however, no functional data address this hypothesis. We examined the relationships between the CYBA C242T polymorphism and direct measurements of superoxide production in human blood vessels. METHODS AND RESULTS: Vascular NAD(P)H oxidase activity was determined in human saphenous veins obtained from 110 patients with coronary artery disease and identified risk factors. Immunoblotting, reverse-transcription polymerase chain reaction, and DNA sequencing showed that p22phox protein, mRNA, and 242C/T allelic variants are expressed in human blood vessels. Vascular superoxide production, both basal and NADH-stimulated, was highly variable between patients, but the presence of the CYBA 242T allele was associated with significantly reduced vascular NAD(P)H oxidase activity, independent of other clinical risk factors for atherosclerosis. CONCLUSIONS: Association of the CYBA 242T allele with reduced NAD(P)H oxidase activity in human blood vessels suggests that genetic variation in NAD(P)H oxidase components may play a significant role in modulating superoxide production in human atherosclerosis. PMID- 11023927 TI - Long-term effects of cholesterol lowering and angiotensin-converting enzyme inhibition on coronary atherosclerosis: The Simvastatin/Enalapril Coronary Atherosclerosis Trial (SCAT). AB - BACKGROUND: This long-term, multicenter, randomized, double-blind, placebo controlled, 2 x 2 factorial, angiographic trial evaluated the effects of cholesterol lowering and angiotensin-converting enzyme inhibition on coronary atherosclerosis in normocholesterolemic patients. METHODS AND RESULTS: There were a total of 460 patients: 230 received simvastatin and 230, a simvastatin placebo, and 229 received enalapril and 231, an enalapril placebo (some subjects received both drugs and some received a double placebo). Mean baseline measurements were as follows: cholesterol level, 5.20 mmol/L; triglyceride level, 1.82 mmol/L; HDL, 0.99 mmol/L; and LDL, 3.36 mmol/L. Average follow-up was 47.8 months. Changes in quantitative coronary angiographic measures between simvastatin and placebo, respectively, were as follows: mean diameters, -0.07 versus -0.14 mm (P:=0.004); minimum diameters, -0.09 versus -0.16 mm (P:=0. 0001); and percent diameter stenosis, 1.67% versus 3.83% (P:=0.0003). These benefits were not observed in patients on enalapril when compared with placebo. No additional benefits were seen in the group receiving both drugs. Simvastatin patients had less need for percutaneous transluminal coronary angioplasty (8 versus 21 events; P:=0.020), and fewer enalapril patients experienced the combined end point of death/myocardial infarction/stroke (16 versus 30; P:=0.043) than their respective placebo patients. CONCLUSIONS: This trial extends the observation of the beneficial angiographic effects of lipid-lowering therapy to normocholesterolemic patients. The implications of the neutral angiographic effects of angiotensin converting enzyme inhibition are uncertain, but they deserve further investigation in light of the positive clinical benefits suggested here and seen elsewhere. PMID- 11023928 TI - Randomized secondary prevention trial of azithromycin in patients with coronary artery disease: primary clinical results of the ACADEMIC study. AB - BACKGROUND: Chlamydia pneumoniae is associated with coronary artery disease (CAD), although its causal role is uncertain. A small preliminary study reported a >50% reduction in ischemic events by azithromycin, an antibiotic effective against C pneumoniae, in seropositive CAD patients. We tested this prospectively in a larger, randomized, double-blind study. METHODS AND RESULTS: CAD patients (n=302) seropositive to C pneumoniae (IgG titers >/=1:16) were randomized to placebo or azithromycin 500 mg/d for 3 days and then 500 mg/wk for 3 months. The primary clinical end point included cardiovascular death, resuscitated cardiac arrest, nonfatal myocardial infarction (MI), stroke, unstable angina, and unplanned coronary revascularization at 2 years. Treatment groups were balanced, and azithromycin was generally well tolerated. During the trial, 47 first primary events occurred (cardiovascular death, 9; resuscitated cardiac arrest, 1; MI, 11; stroke, 3; unstable angina, 4; and unplanned coronary revascularization, 19), with 22 events in the azithromycin group and 25 in the placebo group. There was no significant difference in the 1 primary end point between the 2 groups (hazard ratio for azithromycin, 0.89; 95% CI, 0.51 to 1.61; P:=0.74). Events included 9 versus 7 occurring within 6 months and 13 versus 18 between 6 and 24 months in the azithromycin and placebo groups, respectively. CONCLUSIONS: This study suggests that antibiotic therapy with azithromycin is not associated with marked early reductions (>/=50%) in ischemic events as suggested by an initial published report. However, a clinically worthwhile benefit (ie, 20% to 30%) is still possible, although it may be delayed. Larger (several thousand patient), longer term (>/=3 to 5 years) antibiotic studies are therefore indicated. PMID- 11023929 TI - Survival outcomes 1 year after reperfusion therapy with either alteplase or reteplase for acute myocardial infarction: results from the Global Utilization of Streptokinase and t-PA for Occluded Coronary Arteries (GUSTO) III Trial. AB - BACKGROUND: New recombinant plasminogen activators have been developed to simulate the fibrinolytic action of the physiological serine protease tissue plasminogen activator (alteplase, t-PA), and have prolonged half-life features permitting bolus administration. One such activator, reteplase (r-PA), was compared with t-PA in the Global Utilization of Streptokinase and t-PA for Occluded Coronary Arteries (GUSTO)-III Trial. METHODS AND RESULTS: At 1-year follow-up, survival status was ascertained in 97.4% of the 15 059 patients enrolled in the GUSTO-III trial. At 1 year, the mortality rate for the t-PA assigned group was 11.06%, and for r-PA it was 11.20% (P:=0. 77). The absolute mortality difference of 0.14% has 95% CIs of -1. 21% to 0.93%. There were no significant differences in outcome by intention-to-treat for the 2 different plasminogen activators in the prespecified groups (age, infarct location, time-to treatment). The absolute difference in mortality rates between t-PA and r-PA progressively narrowed over the predetermined observation times after random assignment; it was 0.31% at 24 hours, 0.26% at 7 days, 0.23% at 30 days, and 0.14% at 1 year. Of note, mortality rate in the trial between 30 days and 1 year in 13 883 patients was 4.02% and did not differ between the treatment groups. However, this mortality rate was substantially greater than in GUSTO-I, in which mortality rate for t-PA versus streptokinase between 30 days and 1-year was 2.97% (heart rate 1.36, 95% CI 1.23, 1.50, P:<0.001). CONCLUSIONS: The r-PA and t-PA strategies yielded similar survival outcomes after 30 days in this trial. The increase in mortality rate during extended follow-up compared with previous trials may reflect higher-risk patients and highlights the need for improved secondary prevention strategies. PMID- 11023931 TI - Depressive symptoms and risks of coronary heart disease and mortality in elderly Americans. Cardiovascular Health Study Collaborative Research Group. AB - BACKGROUND: Several epidemiological studies have associated depressive symptoms with cardiovascular disease. We investigated whether depressive symptoms constituted a risk for coronary heart disease (CHD) and total mortality among an apparently healthy elderly cohort. METHODS AND RESULTS: In a prospective cohort of 5888 elderly Americans (>/=65 years) who were enrolled in the Cardiovascular Health Study, 4493 participants who were free of cardiovascular disease at baseline provided annual information on their depressive status, which was assessed using the Depression Scale of the Center for Epidemiological Studies. These 4493 subjects were followed for 6 years for the development of CHD and mortality. The cumulative mean depression score was assessed for each participant up to the time of event (maximum 6-year follow-up). Using time-dependent, proportional-hazards models, the unadjusted hazard ratio associated with every 5 unit increase in mean depression score for the development of CHD was 1.15 (P:=0.006); the ratio for all-cause mortality was 1.29 (P:<0.0001). In multivariate analyses adjusted for age, race, sex, education, diabetes, hypertension, cigarette smoking, total cholesterol, triglyceride level, congestive heart failure, and physical inactivity, the hazard ratio for CHD was 1.15 (P:=0.006) and that for all-cause mortality was 1.16 (P:=0.006). Among participants with the highest cumulative mean depression scores, the risk of CHD increased by 40% and risk of death by 60% compared with those who had the lowest mean scores. CONCLUSIONS: Among elderly Americans, depressive symptoms constitute an independent risk factor for the development of CHD and total mortality. PMID- 11023930 TI - Polyethylene glycol-derivatized cysteine-substitution variants of recombinant staphylokinase for single-bolus treatment of acute myocardial infarction. AB - BACKGROUND: Thrombolytic therapy of acute myocardial infarction (AMI) is evolving toward bolus administration. Derivatization of proteins with polyethylene glycol (PEG) may reduce their clearance. METHODS AND RESULTS: A staphylokinase (SakSTAR) variant with 12 amino acid substitutions to reduce its antigenicity, SakSTAR (K35A, E65Q, K74R, E80A, D82A, T90A, E99D, T101S, E108A, K109A, K130T, K135R), and with Ser in position 3 mutated into Cys (code SY161), was derivatized with maleimide-PEG with M:(r) of 5,000 (P5), 10,000 (P10), or 20,000 (P20). The PEGylated variants recognized only one third of the antibodies elicited with wild type SakSTAR in AMI patients. In experimental animals, plasma clearances were reduced 2. 5- to 5-fold with P5, 5- to 20-fold with P10, and 20-fold with P20, and bolus injection induced pulmonary plasma clot lysis at doses inversely related to their clearance. Intravenous bolus injection of 5 mg of the P5, P10, or P20 variants in AMI patients was associated with plasma half-lives (t(1/2alpha)) of 13, 30, and 120 minutes and clearances of 75, 43, and 8 mL/min, respectively, compared with 3 minutes and 360 mL/min for SakSTAR. Injection of 5 mg P5 variant restored TIMI-3 flow within 60 minutes in 14 of 18 AMI patients (78%, 95% CI 55% to 91%) and of 2.5 mg in 7 of 11 patients (63%, 95% CI 35% to 85%), both in the absence of fibrinogen degradation. The immunogenicity of the variants was significantly (P:<0.002) reduced. CONCLUSIONS: The staphylokinase variant SY161-P5, derivatized with one linear polyethylene glycol molecule of M:(r) 5000, is a promising fibrin-selective agent for single-bolus coronary thrombolysis. PMID- 11023932 TI - Multicenter, randomized, controlled trial of 150-J biphasic shocks compared with 200- to 360-J monophasic shocks in the resuscitation of out-of-hospital cardiac arrest victims. Optimized Response to Cardiac Arrest (ORCA) Investigators. AB - BACKGROUND: In the present study, we compared an automatic external defibrillator (AED) that delivers 150-J biphasic shocks with traditional high-energy (200- to 360-J) monophasic AEDs. METHODS AND RESULTS: AEDs were prospectively randomized according to defibrillation waveform on a daily basis in 4 emergency medical services systems. Defibrillation efficacy, survival to hospital admission and discharge, return of spontaneous circulation, and neurological status at discharge (cerebral performance category) were compared. Of 338 patients with out of-hospital cardiac arrest, 115 had a cardiac etiology, presented with ventricular fibrillation, and were shocked with an AED. The time from the emergency call to the first shock was 8.9+/-3.0 (mean+/-SD) minutes. CONCLUSIONS: The 150-J biphasic waveform defibrillated at higher rates, resulting in more patients who achieved a return of spontaneous circulation. Although survival rates to hospital admission and discharge did not differ, discharged patients who had been resuscitated with biphasic shocks were more likely to have good cerebral performance. PMID- 11023933 TI - Clinical utility of Doppler echocardiography and tissue Doppler imaging in the estimation of left ventricular filling pressures: A comparative simultaneous Doppler-catheterization study. AB - BACKGROUND: Noninvasive assessment of diastolic filling by Doppler echocardiography provides important information about left ventricular (LV) status in selected subsets of patients. This study was designed to assess whether mitral annular velocities as assessed by tissue Doppler imaging are associated with invasive measures of diastolic LV performance and whether additional information is gained over traditional Doppler variables. METHODS AND RESULTS: One hundred consecutive patients referred for cardiac catheterization underwent simultaneous Doppler interrogation. Invasive measurements of LV pressures were obtained with micromanometer-tipped catheters, and the mean LV diastolic pressure (M-LVDP) was used as a surrogate for mean left atrial pressure. Doppler signals from the mitral inflow, pulmonary venous inflow, and TDI of the mitral annulus were obtained. Isolated parameters of transmitral flow correlated with M-LVDP only when ejection fraction <50%. The ratio of mitral velocity to early diastolic velocity of the mitral annulus (E/E') showed a better correlation with M-LVDP than did other Doppler variables for all levels of systolic function. E/E' <8 accurately predicted normal M-LVDP, and E/E' >15 identified increased M-LVDP. Wide variability was present in those with E/E' of 8 to 15. A subset of those patients with E/E' 8 to 15 could be further defined by use of other Doppler data. CONCLUSIONS: The combination of tissue Doppler imaging of the mitral annulus and mitral inflow velocity curves provides better estimates of LV filling pressures than other methods (pulmonary vein, preload reduction). However, accurate prediction of filling pressures for an individual patient requires a stepwise approach incorporating all available data. PMID- 11023934 TI - Independent and incremental prognostic value of (201)Tl lung uptake at rest in patients with severe postischemic left ventricular dysfunction. AB - BACKGROUND: An elevated (201)Tl lung uptake after stress is related to an adverse prognosis. METHODS AND RESULTS: The functional and prognostic significance of resting (201)Tl lung uptake was assessed in 124 consecutive patients with ischemic heart disease and ejection fraction 0.61 best separated patients with and without events (ROC area under curve 0.82). Event-free survival was significantly lower in patients with L/H >0.61 (P:<0. 001); L/H >0.61 (chi(2)=10.8; P:<0.001) and a restrictive filling pattern (chi(2)=3.6; P:<0.05) were independent predictors of events. The prognostic value of L/H was incremental over that obtained by clinical, echographic and Doppler data (global chi(2)=20.8). CONCLUSIONS: In patients with severe postischemic left ventricular dysfunction undergoing rest-redistribution (201)Tl imaging, an increased lung tracer uptake showed incremental prognostic value over clinical and other imaging findings, providing clinically useful risk assessment. PMID- 11023935 TI - Effects of antihypertensive therapy on glucose and insulin metabolism and on left ventricular mass: A randomized, double-blind, controlled study of 21 obese hypertensives. AB - BACKGROUND: Glucose and insulin levels are associated with left ventricular mass (LVM) in insulin-resistant individuals. Antihypertensive drugs have different effects on glucose and insulin metabolism (GIM) and on LVM. To evaluate whether the effects of antihypertensive therapy on LVM are associated with its effects on GIM, we compared the effects of atenolol and perindopril on these parameters in a group of insulin-resistant, obese hypertensives. METHODS AND RESULTS: A total of 21 obese, nondiabetic hypertensives who were aged 55+/-12 years, had a body mass index of 32.8+/-5.0 kg/m(2), were free of coronary or valvular heart disease, and had normal LV function were randomized to treatment with atenolol (n=11) or perindopril (n=10). Echocardiographic LVM corrected for height (LVM/height) and GIM (3-hour intravenous glucose tolerance test) were measured after 4 to 6 weeks of washout and 6 months of treatment. Baseline characteristics were similar in both groups. Atenolol and perindopril effectively reduced blood pressure (from 149+/-13/98+/-4 to 127+/-8/82+/-6 mm Hg and from 148+/-9/98+/-4 to 129+/-9/82+/-6 mm Hg, respectively, for the atenolol and perindopril groups; P:=0.002). Atenolol significantly worsened GIM parameters, fasting glucose levels (5.3+/-0.9 to 6.0+/ 1.5 mmol/L; P:=0.003), fasting insulin levels (121+/-121 to 189+/-228 pmol/L; P:=0.03), and most other relevant metabolic measures (P:<0.05 for all). Perindopril did not affect GIM. Atenolol did not affect LVM/height (119+/-12 to 120+/-17 g/m; P:=0.8), whereas perindopril significantly reduced LVM/height (120+/-13 to 111+/-19 g/m; P:=0.04). CONCLUSIONS: In obese, hypertensive individuals, adequate and similar blood pressure control was achieved with perindopril and atenolol. However, perindopril but not atenolol was associated with a more favorable GIM profile and led to a significant regression of LVM. PMID- 11023936 TI - Electrical remodeling of the atria associated with paroxysmal and chronic atrial flutter. AB - BACKGROUND: Atrial electrical remodeling may be important for the initiation and perpetuation of atrial arrhythmias. Whether paroxysmal atrial flutter (AFL) and chronic AFL cause electrical remodeling of the atria has not been conclusively determined. METHODS AND RESULTS: Before radiofrequency ablation of paroxysmal AFL, 15 patients in sinus rhythm were evaluated under autonomic blockade. Lateral right atrial (LRA) effective refractory periods (ERPs) at 600 and 450 ms were measured before and at 1-minute intervals for 10 minutes after spontaneous or pace termination of a 5- to 10-minute period of induced AFL. In 10 patients with chronic AFL, LRA, septal, and coronary sinus (CS) ERPs and corrected sinus node recovery times (cSNRTs) at 600 and 450 ms were measured under autonomic blockade 15 minutes, 30 minutes, and 3 weeks after termination of chronic AFL by ablation. In the paroxysmal AFL group, LRA ERPs decreased by 18% at 600 ms and 12% at 450 ms (P:<0.01) after induced AFL and recovered to baseline over approximately 5 minutes. Atrial fibrillation developed during AFL in 3 patients and during ERP testing in 3 patients when refractoriness was at its nadir. In the chronic AFL group, LRA, septal, and CS ERPs at 3 weeks were significantly greater than at 15 and 30 minutes after termination of chronic AFL at both cycle lengths (P:<0.01). Three weeks after ablation, cSNRT decreased 35% at 600 ms (P:<0.05) and decreased 44% at 450 ms (P:<0. 05). Both ERPs and cSNRTs measured 15 and 30 minutes after ablation of chronic AFL were not significantly different. CONCLUSIONS: Both paroxysmal AFL and chronic AFL cause reversible electrical remodeling of the atria but demonstrate different time courses of recovery. PMID- 11023937 TI - Both beta(2)- and beta(1)-adrenergic receptors mediate hastened relaxation and phosphorylation of phospholamban and troponin I in ventricular myocardium of Fallot infants, consistent with selective coupling of beta(2)-adrenergic receptors to G(s)-protein. AB - BACKGROUND: In adult human heart, both beta(1)- and beta(2)-adrenergic receptors mediate hastening of relaxation; however, it is unknown whether this also occurs in infant heart. We compared the effects of stimulation of beta(1)- and beta(2) adrenergic receptors on relaxation and phosphorylation of phospholamban and troponin I in ventricle obtained from infants with tetralogy of Fallot. METHODS AND RESULTS: Myocardium dissected from the right ventricular outflow tract of 27 infants (age range 21/2 to 35 months) with tetralogy of Fallot was set up to contract 60 times per minute. Selective stimulation of beta(1)-adrenergic receptors with (-)-norepinephrine (NE) and beta(2)-adrenergic receptors with (-) epinephrine (EPI) evoked phosphorylation of phospholamban (at serine-16 and threonine-17) and troponin I and caused concentration-dependent increases in contractile force (-log EC(50) [mol/L] NE 5.5+/-0.1, n=12; EPI 5.6+/-0.1, n=13 patients), hastening of the time to reach peak force (-log EC(50) [mol/L] NE 5.8+/-0.2; EPI 5.8+/-0.2) and 50% relaxation (-log EC(50) [mol/L] NE 5.7+/-0.2; EPI 5.8+/-0.1). Ventricular membranes from Fallot infants, labeled with (-) [(125)I]-cyanopindolol, revealed a greater percentage of beta(1)- (71%) than beta(2)-adrenergic receptors (29%). Binding of (-)-epinephrine to beta(2) receptors underwent greater GTP shifts than binding of (-)-norepinephrine to beta(1)-receptors. CONCLUSIONS: Despite their low density, beta(2)-adrenergic receptors are nearly as effective as beta(1)-adrenergic receptors of infant Fallot ventricle in enhancing contraction, relaxation, and phosphorylation of phospholamban and troponin I, consistent with selective coupling to G(s)-protein. PMID- 11023938 TI - Adoptive transfer of beta(2)-glycoprotein I-reactive lymphocytes enhances early atherosclerosis in LDL receptor-deficient mice. AB - BACKGROUND: It has been proposed that autoimmune factors can influence the progression of atherosclerosis. We have previously shown that immunization of LDL receptor-deficient (LDL-RD mice) with beta(2)-glycoprotein I (beta2GPI; a principal target of "autoimmune" antiphospholipid antibodies) enhances early atherosclerosis. In the present study, we tested the hypothesis that adoptive transfer of beta2GPI-reactive T cells can accelerate fatty streak formation in LDL-RD mice. METHODS AND RESULTS: LDL-RD mice were immunized with human beta2GPI. An additional group of mice were immunized with beta2GPI and boosted with the same antigen 3 weeks later. Control mice with immunized with human serum albumin. Lymphocytes obtained from the draining lymph node cells or from splenocytes of beta2GPI- or human serum albumin-immunized mice were stimulated in vitro with beta2GPI or with the mitogen concavalin A, respectively. The cultured lymphocytes were transferred intraperitoneally to syngenic LDL-RD mice, and the mice were fed a high-fat "Western" diet for 5 weeks until death. Mice injected with lymphocytes from draining lymph nodes or spleens of beta2GPI-immunized animals displayed larger fatty streaks than those induced by control treated animals. T-cell depleted splenocytes from beta2GPI were unable to promote lesion formation in the mice. CONCLUSIONS: The present study provides the first direct evidence for a role of antigen (beta2GPI)-reactive T cells in the promotion of fatty streaks in mice. PMID- 11023939 TI - Differential effects of estrogen and progesterone on AT(1) receptor gene expression in vascular smooth muscle cells. AB - BACKGROUND: The beneficial vasoprotective effects of a postmenopausal estrogen replacement therapy may be prevented by a concomitant administration of progestins. To investigate the differential effects of estrogens and progesterone, we examined their influence on AT(1) receptor gene expression in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: 17beta-Estradiol caused downregulation of AT(1) receptor mRNA expression to 46+/-14%, whereas progesterone led to a significant upregulation to 201+/-29%, as assessed by Northern analysis. Western blots revealed that estrogen induced a downregulation and progesterone an upregulation of the AT(1) receptor protein. Estrogen-induced decrease of AT(1) receptor expression was mediated through activation of estrogen receptors. Nuclear run-on assays revealed that 17beta-estradiol did not alter AT(1) receptor mRNA transcription rate, whereas progesterone caused an enhanced AT(1) receptor mRNA transcription rate. 17beta-Estradiol decreased the AT(1) receptor mRNA half-life from 5 to 2 hours, whereas progesterone induced a stabilization of AT(1) receptor mRNA to a half-life of 10 hours. Preincubation of VSMCs with PD98059, SB203580, herbimycin, wortmannin, or N:(omega)-nitro-L arginine suggested that 17beta-estradiol caused AT(1) receptor downregulation through nitric oxide-dependent pathways. Progesterone caused AT(1) receptor overexpression via PI(3)-kinase activation. Angiotensin II-induced release of reactive oxygen species was inhibited by estrogens. Progesterone itself enhanced the production of reactive oxygen species. CONCLUSIONS: Because AT(1) receptor regulation plays a pivotal role in the pathogenesis of hypertension and atherosclerosis, the differential effects of estrogen and progesterone on the expression of this gene may in part explain the potentially counteracting effects of these reproductive hormones on the incidence of postmenopausal cardiovascular diseases. PMID- 11023940 TI - Peroxisome proliferator-activated receptor gamma activators downregulate angiotensin II type 1 receptor in vascular smooth muscle cells. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPARgamma) activators, such as troglitazone (Tro), not only improve insulin resistance but also suppress the neointimal formation after balloon injury. However, the precise mechanisms have not been determined. Angiotensin II (Ang II) plays crucial roles in the pathogenesis of atherosclerosis, hypertension, and neointimal formation after angioplasty. We examined the effect of PPARgamma activators on the expression of Ang II type 1 receptor (AT(1)-R) in cultured vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: AT(1)-R mRNA and AT(1)-R protein levels were determined by Northern blot analysis and radioligand binding assay, respectively. Natural PPARgamma ligand 15-deoxy-Delta(12,14)-prostaglandin J(2), as well as Tro, reduced the AT(1)-R mRNA expression and the AT(1)-R protein level. The PPARgamma activators also reduced the calcium response of VSMCs to Ang II. PPARgamma activators suppressed the AT(1)-R promoter activity measured by luciferase assay but did not affect the AT(1)-R mRNA stability, suggesting that the suppression occurs at the transcriptional level. CONCLUSIONS: PPARgamma activators reduced the AT(1)-R expression and calcium response to Ang II in VSMCs. Downregulation of AT(1)-R may contribute to the inhibition of neointimal formation by PPARgamma activators. PMID- 11023941 TI - Myocyte response to beta-adrenergic stimulation is preserved in the noninfarcted myocardium of globally dysfunctional rat hearts after myocardial infarction. AB - BACKGROUND: Cellular mechanisms underlying the diminished inotropic response of remodeled hearts after myocardial infarction (MI) remain unclear. METHODS AND RESULTS: Left ventricular (LV) remodeling and function were assessed by 2D echocardiography and isolated perfused heart studies in 6-week post-MI and sham operated rats. LV myocytes from sham and noninfarcted MI hearts were used for morphometric and functional studies. Beta-adrenergic receptor (beta-AR) agonist isoproterenol (ISO)-induced contractile response was measured in isolated hearts. The effects of ISO and forskolin on contractile function and calcium transients of isolated myocytes were recorded. ISO-induced cAMP generation was compared in sham and MI myocytes. beta-AR density was measured by radioligand binding. MI hearts were remodeled (LV diameter 8.5+/-0.3 versus 5.7+/-0.3 mm, P:<0.001) and showed global (% fractional shortening 19.1+/-2.5 versus 55.3+/-2.2, P:<0.01) and regional contractile dysfunction of noninfarcted myocardium (% systolic posterior wall thickening 37+/-4 versus 62+/-10, P:<0.01). Isolated heart function (LV developed pressure 58+/-2 versus 72+/-3 mm Hg, P:=0.004) and ISO concentration response were reduced in MI hearts. Myocytes from the noninfarcted LV were structurally remodeled (32% longer and 18% wider), but their contractile response and intracellular calcium kinetics to ISO and forskolin were not diminished. beta AR receptor density (B(max) 24+/-1.5 versus 22.4+/-1.6 fmol/mg protein) and beta AR agonist-stimulated cAMP were similar in both groups. CONCLUSIONS: Isolated myocytes from the remodeled and dysfunctional myocardium are structurally modified but contract normally under basal conditions and in response to beta-AR stimulation. beta-AR density is preserved in remodeled myocytes. Nonmyocyte factors may be more important in the genesis of contractile dysfunction in the remodeled rat heart up to 6 weeks after MI. PMID- 11023942 TI - Heart failure affects mitochondrial but not myofibrillar intrinsic properties of skeletal muscle. AB - BACKGROUND: Congestive heart failure (CHF) induces abnormalities in skeletal muscle that are thought to in part explain exercise intolerance. The aim of the present study was to determine whether these changes actually result in contractile or metabolic functional alterations and whether they are muscle type specific. METHODS AND RESULTS: With a rat model of CHF (induced by aortic banding), we studied mitochondrial function, mechanical properties, and creatine kinase (CK) compartmentation in situ in permeabilized fibers from soleus (SOL), an oxidative slow-twitch muscle, and white gastrocnemius (GAS), a glycolytic fast twitch muscle. Animals were studied 7 months after surgery, and CHF was documented on the basis of anatomic data. Alterations in skeletal muscle phenotype were documented with an increased proportion of fast-type fiber and fast myosin heavy chain, decreased capillary-to-fiber ratio, and decreased citrate synthase activity. Despite a slow-to-fast phenotype transition in SOL, no change was observed in contractile capacity or calcium sensitivity. However, muscles from CHF rats exhibited a dramatic decrease in oxidative capacities (oxygen consumption per gram of fiber dry weight) of 35% for SOL and 45% for GAS (P:<0.001). Moreover, the regulation of respiration with ADP and mitochondrial CK and adenylate kinase was impaired in CHF SOL. Mitochondrial CK activity and content (Western blots) were dramatically decreased in both muscles. CONCLUSIONS: CHF results in alterations in both mitochondrial function and phosphotransfer systems but unchanged myofibrillar function in skeletal muscles, which suggests a myopathy of metabolic origin in CHF. PMID- 11023943 TI - Chronic exercise reduces sympathetic nerve activity in rabbits with pacing induced heart failure: A role for angiotensin II. AB - BACKGROUND: Chronic exercise (EX) improves the quality of life and increases the survival of patients with chronic heart failure (CHF). Because sympathetic nerve activity is elevated in the CHF state, it is possible that EX is beneficial in this disease due to a decrease in sympathetic outflow. METHODS AND RESULTS: We evaluated arterial baroreflex function and resting renal sympathetic nerve activity (RSNA) in EX normal and CHF rabbits before and after angiotensin II type 1 (AT(1)) receptor blockade. Four groups of rabbits were studied: a normal non-EX group, a normal EX group, a CHF non-EX group, and a CHF EX group. EX lowered resting RSNA in rabbits with CHF but not in normal rabbits. In addition, EX increased arterial baroreflex sensitivity in the CHF group (heart rate slope: CHF 1. 7+/-0.3 bpm/mm Hg, EX CHF 4.9+/-0.3 bpm/mm Hg; P:<0.01; RSNA slope: CHF 2.2+/ 0.2%max/mm Hg, EX CHF 5.7+/-0.4%max/mm Hg; P:<0.01. AT(1) receptor blockade enhanced baroreflex sensitivity in the non-EX CHF rabbits but had no effect in EX CHF rabbits. Concomitant with this effect, EX lowered the elevated plasma angiotensin II concentration in the CHF group. A significant positive correlation was observed between sympathetic nerve activity and plasma angiotensin II. CONCLUSIONS: These data strongly suggest that EX reduces the sympathoexcitatory state in the setting of CHF. Enhanced arterial baroreflex sensitivity may contribute to this reduction. In addition, EX lowers plasma angiotensin II concentration in CHF. These data further suggest that the lowering of angiotensin II may contribute to the decrease in sympathetic nerve activity after EX in the CHF state. PMID- 11023944 TI - Spontaneously occurring arrhythmogenic right ventricular cardiomyopathy in the domestic cat: A new animal model similar to the human disease. AB - BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a primary myocardial disease of incompletely resolved pathogenesis and is a largely unappreciated cause of sudden death in the young. METHODS AND RESULTS: Clinical features of 12 domestic cats with ARVC (7 male; 1 to 20 years old, mean 7.3+/-5.2 years) were right-sided congestive heart failure (n=8), supraventricular tachyarrhythmias (n=5), ventricular tachycardia (n=3), polymorphic ventricular arrhythmias (n=6), and right bundle-branch block (n=5). ARVC was suspected in all 8 cats examined with echocardiography by marked enlargement of the right ventricle (RV) and right atrium and tricuspid regurgitation. Eight died of cardiovascular disease and 4 died of noncardiac conditions. At autopsy, hearts of ARVC cats were characterized grossly by moderate-to-severe RV cavity enlargement and wall thinning (n=12) and apical aneurysm formation (n=6). Histology demonstrated pronounced RV lesions in all 12 ARVC cats, including marked myocardial injury (myocyte death and atrophy) and repair (fibrous and/or fatty replacement). Injury and repair were also evident in the left ventricle (LV) in 10 cats, and 2 had involvement of both atria. Myocarditis was present in 10 of the 12 ARVC cats. Apoptosis was detected in 9 ARVC cats (mean apoptotic index, 28+/-23% RV, 21+/-19% LV, and 17+/-15% ventricular septum) but not in controls. CONCLUSIONS: In the common domestic cat, we identified a clinically relevant cardiomyopathy that closely mimics ARVC in humans. This unique feline model of human disease will be relevant to defining pathogenesis and investigating mechanisms responsible for disease progression in ARVC. PMID- 11023945 TI - Multislice spiral CT follow-up of a patient with implanted DeBakey ventricular assist device. PMID- 11023946 TI - Ultrasound finding of a mobile atheroma in the common carotid artery. PMID- 11023947 TI - Simple index for prediction of cardiac risk in stable patients undergoing non urgent major noncardiac surgery: what about the more severely compromised patients? PMID- 11023948 TI - Left ventricular performance in ischemic right ventricular dysfunction. PMID- 11023949 TI - sFlt-1, a potential antagonist for exogenous VEGF. PMID- 11023950 TI - Acute platelet inhibition with abciximab does not reduce in-stent restenosis. PMID- 11023951 TI - Acute platelet inhibition with abciximab does not reduce in-stent restenosis. PMID- 11023952 TI - Uninsured numbers drop. PMID- 11023953 TI - Relationships between enterococcal virulence and antimicrobial resistance. AB - Enterococci have become a vexing problem in clinical medicine because of their ability to infect patients who are typically receiving antibiotic therapy for unrelated underlying illness. Moreover, the infections have become extremely difficult to manage because of the accumulation of antibiotic resistances among enterococci. The ability of enterococci to cause disease is an intrinsic property of the organism or possibly subpopulations within enterococcal species. The probability of an infection's becoming established, however, is almost certainly in part a function of the enterococcal burden. By altering endogenous bacterial flora, antibiotic therapy promotes increased colonization by antibiotic-resistant organisms. Therefore, antibiotic resistance and intrinsic virulence both contribute to disease, but in separate and complementary ways. We review the virulence of enterococci, as distinct from the acquisition of antimicrobial resistance genes, and identify current gaps in our understanding of enterococcal virulence and the basis for disease. PMID- 11023954 TI - Polysaccharide immunomodulators as therapeutic agents: structural aspects and biologic function. AB - Polysaccharide immunomodulators were first discovered over 40 years ago. Although very few have been rigorously studied, recent reports have revealed the mechanism of action and structure-function attributes of some of these molecules. Certain polysaccharide immunomodulators have been identified that have profound effects in the regulation of immune responses during the progression of infectious diseases, and studies have begun to define structural aspects of these molecules that govern their function and interaction with cells of the host immune system. These polymers can influence innate and cell-mediated immunity through interactions with T cells, monocytes, macrophages, and polymorphonuclear lymphocytes. The ability to modulate the immune response in an appropriate way can enhance the host's immune response to certain infections. In addition, this strategy can be utilized to augment current treatment regimens such as antimicrobial therapy that are becoming less efficacious with the advent of antibiotic resistance. This review focuses on recent studies that illustrate the structural and biologic activities of specific polysaccharide immunomodulators and outlines their potential for clinical use. PMID- 11023955 TI - Classification, identification, and clinical significance of Proteus, Providencia, and Morganella. AB - This review presents the current taxonomy of the genera Proteus, Providencia, and Morganella, along with the current methods for the identification of each species within the three genera, incorporating both conventional biochemical and commercial methods. While all of these organisms are ubiquitous in the environment, individual case reports and nosocomial outbreak reports that demonstrate their ability to cause major infectious disease problems are presented. Lastly, anticipated antimicrobial susceptibility patterns are reviewed. Many of these organisms are easily controlled, but the advent of newer and more powerful antimicrobial agents has led to some problems of which laboratorians need to be aware. PMID- 11023956 TI - Systemic diseases caused by oral infection. AB - Recently, it has been recognized that oral infection, especially periodontitis, may affect the course and pathogenesis of a number of systemic diseases, such as cardiovascular disease, bacterial pneumonia, diabetes mellitus, and low birth weight. The purpose of this review is to evaluate the current status of oral infections, especially periodontitis, as a causal factor for systemic diseases. Three mechanisms or pathways linking oral infections to secondary systemic effects have been proposed: (i) metastatic spread of infection from the oral cavity as a result of transient bacteremia, (ii) metastatic injury from the effects of circulating oral microbial toxins, and (iii) metastatic inflammation caused by immunological injury induced by oral microorganisms. Periodontitis as a major oral infection may affect the host's susceptibility to systemic disease in three ways: by shared risk factors; subgingival biofilms acting as reservoirs of gram-negative bacteria; and the periodontium acting as a reservoir of inflammatory mediators. Proposed evidence and mechanisms of the above odontogenic systemic diseases are given. PMID- 11023957 TI - Multiplex PCR: optimization and application in diagnostic virology. AB - PCR has revolutionized the field of infectious disease diagnosis. To overcome the inherent disadvantage of cost and to improve the diagnostic capacity of the test, multiplex PCR, a variant of the test in which more than one target sequence is amplified using more than one pair of primers, has been developed. Multiplex PCRs to detect viral, bacterial, and/or other infectious agents in one reaction tube have been described. Early studies highlighted the obstacles that can jeopardize the production of sensitive and specific multiplex assays, but more recent studies have provided systematic protocols and technical improvements for simple test design. The most useful of these are the empirical choice of oligonucleotide primers and the use of hot start-based PCR methodology. These advances along with others to enhance sensitivity and specificity and to facilitate automation have resulted in the appearance of numerous publications regarding the application of multiplex PCR in the diagnosis of infectious agents, especially those which target viral nucleic acids. This article reviews the principles, optimization, and application of multiplex PCR for the detection of viruses of clinical and epidemiological importance. PMID- 11023958 TI - Rubella virus replication and links to teratogenicity. AB - Rubella virus (RV) is the causative agent of the disease known more popularly as German measles. Rubella is predominantly a childhood disease and is endemic throughout the world. Natural infections of rubella occur only in humans and are generally mild. Complications of rubella infection, most commonly polyarthralgia in adult women, do exist; occasionally more serious sequelae occur. However, the primary public health concern of RV infection is its teratogenicity. RV infection of women during the first trimester of pregnancy can induce a spectrum of congenital defects in the newborn, known as congenital rubella syndrome (CRS). The development of vaccines and implementation of vaccination strategies have substantially reduced the incidence of disease and in turn of CRS in developed countries. The pathway whereby RV infection leads to teratogenesis has not been elucidated, but the cytopathology in infected fetal tissues suggests necrosis and/or apoptosis as well as inhibition of cell division of critical precursor cells involved in organogenesis. In cell culture, a number of unusual features of RV replication have been observed, including mitochondrial abnormalities, and disruption of the cytoskeleton; these manifestations are most probably linked and play some role in RV teratogenesis. Further understanding of the mechanism of RV teratogenesis will be brought about by the investigation of RV replication and virus-host interactions. PMID- 11023959 TI - Limiting the spread of resistant pneumococci: biological and epidemiologic evidence for the effectiveness of alternative interventions. AB - Streptococcus pneumoniae infections are a leading cause of respiratory illness in young children, the elderly, and persons with chronic medical conditions. The emergence of multidrug-resistant pneumococci has compromised the effectiveness of antibiotic therapy for pneumococcal infections. As antibiotic-resistant strains increase in prevalence, there is a need for interventions that minimize the spread of resistant pneumococci. In this review we provide a framework for understanding the spread of pneumococcal resistance and evaluate proposed interventions to reduce this spread. Pneumococci differ from many drug-resistant pathogens because asymptomatic carriers play a key role in transmission of resistant strains and the genes encoding resistance are spread primarily by transformation and conjugative transposons. Evidence suggests that modifications of treatment regimens that have proved effective at limiting resistance in other pathogens may not prevent the spread of pneumococcal resistance. In contrast, programs encouraging more judicious antibiotic use have been shown to be effective. Additionally, a newly developed conjugate pneumococcal vaccine holds great potential as an "antiresistance vaccine" that simultaneously reduces the burden of invasive disease and the prevalence of resistant strains. Several areas of future epidemiologic and laboratory research hold promise to contribute to the reduced spread of pneumococcal resistance. PMID- 11023960 TI - Passive immunity in prevention and treatment of infectious diseases. AB - Antibodies have been used for over a century in the prevention and treatment of infectious disease. They are used most commonly for the prevention of measles, hepatitis A, hepatitis B, tetanus, varicella, rabies, and vaccinia. Although their use in the treatment of bacterial infection has largely been supplanted by antibiotics, antibodies remain a critical component of the treatment of diptheria, tetanus, and botulism. High-dose intravenous immunoglobulin can be used to treat certain viral infections in immunocompromised patients (e.g., cytomegalovirus, parvovirus B19, and enterovirus infections). Antibodies may also be of value in toxic shock syndrome, Ebola virus, and refractory staphylococcal infections. Palivizumab, the first monoclonal antibody licensed (in 1998) for an infectious disease, can prevent respiratory syncytial virus infection in high risk infants. The development and use of additional monoclonal antibodies to key epitopes of microbial pathogens may further define protective humoral responses and lead to new approaches for the prevention and treatment of infectious diseases. PMID- 11023962 TI - Molecular strategies for interrupting arthropod-borne virus transmission by mosquitoes. AB - Arthropod-borne virus (arbovirus) infections cause a number of emerging and resurgent human and veterinary infectious diseases. Traditional means of controlling arbovirus diseases include vaccination of susceptible vertebrates and mosquito control, but in many cases these have been unavailable or ineffective, and so novel strategies for disease control are needed. One possibility is genetic manipulation of mosquito vectors to render them unable to transmit arboviruses. This review describes recent work to test the concept of pathogen derived resistance in arthropods by expression of viral genes in mosquito cell cultures and mosquitoes. Sense and antisense genome sequences from La Crosse virus (LAC) (a member of the Bunyaviridae) and dengue viruses serotypes 1 to 4 (DEN-1 to DEN-4) (members of the Flaviviridae) were expressed in mosquito cells from double-subgenomic and replicon vectors based on Sindbis virus (a member of the Togaviridae). The cells were then challenged with homologous or related viruses. For LAC, expression of antisense sequences from the small (S) genome segment, particularly full-length antisense S RNA, effectively interfered with replication of challenge virus, whereas expression of either antisense or sense RNA from the medium (M) segment was completely ineffective in LAC inhibition. Expression of sense and antisense RNA derived from certain regions of the DEN genome also blocked homologous virus replication more effectively than did RNA from other regions. Other parameters of RNA-mediated interference have been defined, such as the time when replication is blocked and the minimum size of effector RNA. The mechanism of RNA inhibition has not been determined, although it resembles double-stranded RNA interference in other nonvertebrate systems. Prospects for application of molecular strategies to control arbovirus diseases are briefly reviewed. PMID- 11023961 TI - Interference of antibacterial agents with phagocyte functions: immunomodulation or "immuno-fairy tales"? AB - Professional phagocytes (polymorphonuclear neutrophils and monocytes/macrophages) are a main component of the immune system. These cells are involved in both host defenses and various pathological settings characterized by excessive inflammation. Accordingly, they are key targets for immunomodulatory drugs, among which antibacterial agents are promising candidates. The basic and historical concepts of immunomodulation will first be briefly reviewed. Phagocyte complexity will then be unravelled (at least in terms of what we know about the origin, subsets, ambivalent roles, functional capacities, and transductional pathways of this cell and how to explore them). The core subject of this review will be the many possible interactions between antibacterial agents and phagocytes, classified according to demonstrated or potential clinical relevance (e.g., neutropenia, intracellular accumulation, and modulation of bacterial virulence). A detailed review of direct in vitro effects will be provided for the various antibacterial drug families, followed by a discussion of the clinical relevance of these effects in two particular settings: immune deficiency and inflammatory diseases. The prophylactic and therapeutic use of immunomodulatory antibiotics will be considered before conclusions are drawn about the emerging (optimistic) vision of future therapeutic prospects to deal with largely unknown new diseases and new pathogens by using new agents, new techniques, and a better understanding of the phagocyte in particular and the immune system in general. PMID- 11023966 TI - Reversible excitation light-induced enhancement of fluorescence of live mammalian mitochondria. AB - During continuous irradiation with near-ultraviolet light (l = 36510 nm; 16 mW/mm(2)) for 2-3 min, live mammalian cells increased reversibly the intensity of one or more peaks of their autofluorescence spectrum from an initial ('ground') level to a two- to threefold higher ('active') level. The effect is characterized by the existence of two states of quantum efficiency and a mechanism of transition that expresses a threshold and a refractory period. It appears that mitochondria are the principal sources of the rising autofluorescence intensity; however, not all mitochondria are capable of expressing it. Studying cells from various organisms that belong to various branches of the phylogenetic tree, we found the rapid increase of autofluorescence only in placental mammalian cells. We speculate that the effect may point to the ability of placental mammalian mitochondria to generate pulsating light signals. PMID- 11023965 TI - Pathogenic roles for fungal melanins. AB - Melanins represent virulence factors for several pathogenic fungi; the number of examples is growing. Thus, albino mutants of several genera (in one case, mutated precisely in the melanizing enzyme) exhibit decreased virulence in mice. We consider the phenomenon in relation to known chemical properties of melanin, beginning with biosynthesis from ortho-hydroquinone precursors which, when oxidized enzymatically to quinones, polymerize spontaneously to melanin. It follows that melanizing intermediates are cross-linking reagents; melanization stabilizes the external cell wall against hydrolysis and is thought to determine semipermeability in the osmotic ram (the appressorium) of certain plant pathogens. Polymeric melanins undergo reversible oxidation-reduction reactions between cell wall-penetrating quinone and hydroquinone oxidation states and thus represent polymeric redox buffers; using strong oxidants, it is possible to titrate the melanin on living cells and thereby demonstrate protection conferred by melanin in several species. The amount of buffering per cell approximately neutralizes the amount of oxidant generated by a single macrophage. Moreover, the intermediate oxidation state, the semiquinone, is a very stable free radical and is thought to trap unpaired electrons. We have suggested that the oxidation state of external melanin may be regulated by external Fe(II). An independent hypothesis holds that in Cryptococcus neoformans, an important function of the melanizing enzyme (apart from melanization) is the oxidation of Fe(II) to Fe(III), thereby forestalling generation of the harmful hydroxyl radical from H(2)O(2). Thus, problems in fungal pathogenesis have led to evolving hypotheses regarding melanin functioning. PMID- 11023967 TI - Reduced cardiac hypertrophy and altered blood pressure control in transgenic rats with the human tissue kallikrein gene. AB - To evaluate the cardiovascular actions of kinins, we established a transgenic rat line harboring the human tissue kallikrein gene, TGR(hKLK1). Under the control of the zinc-inducible metallothionein promoter, the transgene was expressed in most tissues including the heart, kidney, lung, and brain, and human kallikrein was detected in the urine of transgenic animals. Transgenic rats had a lower 24-h mean arterial pressure in comparison with control rats, which was further decreased when their diet was supplemented with zinc. The day/night rhythm of blood pressure was significantly diminished in TGR(hKLK1) animals, whereas the circadian rhythms of heart rate and locomotor activity were unaffected. Induction of cardiac hypertrophy by isoproterenol treatment revealed a marked protective effect of the kallikrein transgene because the cardiac weight of TGR(hKLK1) increased significantly less, and the expression of atrial natriuretic peptide and collagen III as markers for hypertrophy and fibrosis, respectively, were less enhanced. The specific kinin-B2 receptor antagonist, icatibant, abolished this cardioprotective effect. In conclusion, the kallikrein-kinin system is an important determinant in the regulation of blood pressure and its circadian rhythmicity. It also exerts antihypertrophic and antifibrotic actions in the heart. PMID- 11023964 TI - Vancomycin-resistant enterococci. AB - After they were first identified in the mid-1980s, vancomycin-resistant enterococci (VRE) spread rapidly and became a major problem in many institutions both in Europe and the United States. Since VRE have intrinsic resistance to most of the commonly used antibiotics and the ability to acquire resistance to most of the current available antibiotics, either by mutation or by receipt of foreign genetic material, they have a selective advantage over other microorganisms in the intestinal flora and pose a major therapeutic challenge. The possibility of transfer of vancomycin resistance genes to other gram-positive organisms raises significant concerns about the emergence of vancomycin-resistant Staphylococcus aureus. We review VRE, including their history, mechanisms of resistance, epidemiology, control measures, and treatment. PMID- 11023968 TI - Increased kallikrein expression protects against cardiac ischemia. AB - Multiple indirect lines of evidence point at a cardioprotective role for enhanced bradykinin formation. In particular, the inhibition of angiotensin-converting enzyme, also known as kininase II, can protect against cardiac ischemia, putatively via accumulation of bradykinin. To address whether an increase in kinin formation is sufficient to protect against cardiac ischemia, we studied transgenic rats harboring the human tissue kallikrein gene TGR(hKLK1) under the control of the metallothionein promoter, which drives expression of the transgene in various organs including the heart. We subjected the isolated hearts from transgenic rats and their transgene negative littermates to ex vivo regional cardiac ischemia and reperfusion. During the experiment, the hearts were treated with either vehicle or the specific bradykinin type 2 receptor antagonist HOE 140 (10-9 M). In the transgenic rats, overflow of nucleotide breakdown products upon reperfusion was significantly less (455 +-54 nmol/min/g in transgene negative rats vs. 270+-57 nmol/min/g in the transgenic rats, P. PMID- 11023963 TI - Fungal and parasitic infections of the eye. AB - The unique structure of the human eye as well as exposure of the eye directly to the environment renders it vulnerable to a number of uncommon infectious diseases caused by fungi and parasites. Host defenses directed against these microorganisms, once anatomical barriers are breached, are often insufficient to prevent loss of vision. Therefore, the timely identification and treatment of the involved microorganisms are paramount. The anatomy of the eye and its surrounding structures is presented with an emphasis upon the association of the anatomy with specific infection of fungi and parasites. For example, filamentous fungal infections of the eye are usually due to penetrating trauma by objects contaminated by vegetable matter of the cornea or globe or, by extension, of infection from adjacent paranasal sinuses. Fungal endophthalmitis and chorioretinitis, on the other hand, are usually the result of antecedent fungemia seeding the ocular tissue. Candida spp. are the most common cause of endogenous endophthalmitis, although initial infection with the dimorphic fungi may lead to infection and scarring of the chorioretina. Contact lens wear is associated with keratitis caused by yeasts, filamentous fungi, and Acanthamoebae spp. Most parasitic infections of the eye, however, arise following bloodborne carriage of the microorganism to the eye or adjacent structures. PMID- 11023969 TI - Lipocortin 1 reduces myocardial ischemia-reperfusion injury by affecting local leukocyte recruitment. AB - We assessed here the effect of the glucocorticoid-regulated protein lipocortin 1 (LC1) in a model of rat myocardial ischemia reperfusion. Treatment of animals with human recombinant LC1 at the end of a 25-min ischemic period significantly reduced the extent of infarct size in the area at risk as measured 2 h later, with approximately 50% inhibition at the highest dose tested of 50 microg per rat (equivalent to 5.4 nmol/kg). The protective effect of LC1 was abolished by protein denaturation and not mimicked by the structurally related protein annexin V. A combination of electron and light microscopy techniques demonstrated the occurrence of the myocardial damage at the end of the reperfusion period, with loss of fiber organization. LC1 provided a partial and visible protection. The dose-dependent protection afforded by LC1 was paralleled by lower values of myeloperoxidase activity, tumor necrosis factor a, and macrophage inflammatory protein-1a. The functional link between migrated leukocytes and the myocardial damage was confirmed by electron and light microscopy, and a significantly lower number of extravasated leukocytes was counted in the group of rats treated with LC1 (50 microg). In conclusion, we demonstrate for the first time that LC1 reduces the leukocyte-dependent myocardial damage associated with an ischemia reperfusion procedure. PMID- 11023970 TI - The Egr-1 promoter contains information for constitutive and inducible expression in transgenic mice. AB - Egr-1 is an immediate early gene that couples short-term changes in the extracellular milieu to long-term changes in gene expression. Under in vitro conditions, the Egr-1 gene is expressed in many cell types and is induced by a wide variety of extracellular signals. The mechanisms by which the Egr-1 gene is regulated in vivo remain poorly understood. In this study, we have generated transgenic mice with a construct containing 1200 bp of the mouse Egr-1 promoter coupled to nuclear localized LacZ. In multiple independent lines of mice, reporter gene expression was detected in subsets of endothelial cells, vascular smooth-muscle cells, cardiomyocytes, neurons, and hepatocytes. This pattern closely resembled that of the endogenous gene. After partial hepatectomy, reporter gene activity was upregulated between two- and fivefold in regenerating livers. Taken together, these findings suggest that the Egr-1 promoter contains information for appropriate spatial and temporal expression in vivo. PMID- 11023971 TI - Changes in the balance between mitogenic and antimitogenic lipid second messengers during proliferation, cell arrest, and apoptosis in T-lymphocytes. AB - Control of lymphocyte cell survival and proliferation is critical for both the immune response and for the prevention of autoimmune and infectious diseases. The actions of interleukin-2, the major T-cell regulatory cytokine, are mediated by the complex network of divergent signalling pathways controlled by its high affinity receptor. Various studies have indicated that the generation of certain lipid second messengers is an important mechanism in the control of proliferation and cell death. We have examined the relationship between diacylglycerol and ceramide and the levels of the lipids phosphatidylcholine and sphingomyelin, their potential precursors, in the human T-cell line Kit 225 cultured in three distinct conditions to favor apoptosis, cell arrest, and proliferation. Our data show that, in proliferating cells, the ratios of diacylglycerol/ceramide and phosphatidylcholine/sphingomyelin are higher than those found in arrested cells and increase with time in culture. These ratios are rapidly reversed in apoptotic cells. Further experiments reveal that de novo synthesis of both diacylglycerol and phosphatidylcholine is greatest in proliferating cells, whereas sphingomyelin synthase activity is increased in cells undergoing apoptosis. In summary, our results demonstrate for the first time that the ratio of mitogenic/antimitogenic lipids changes dramatically during T-cell proliferation and cell death. These results indicate that lipid second messengers and the enzymes that are responsible for their generation may provide targets for novel therapeutic interventions in the clinical management of immunosuppression and autoimmune disease. PMID- 11023972 TI - Co-repressors 2000. AB - In the last 5 years, many co-repressors have been identified in eukaryotes that function in a wide range of species, from yeast to Drosophila and humans. Co repressors are coregulators that are recruited by DNA-bound transcriptional silencers and play essential roles in many pathways including differentiation, proliferation, programmed cell death, and cell cycle. Accordingly, it has been shown that aberrant interactions of co-repressors with transcriptional silencers provide the molecular basis of a variety of human diseases. Co-repressors mediate transcriptional silencing by mechanisms that include direct inhibition of the basal transcription machinery and recruitment of chromatin-modifying enzymes. Chromatin modification includes histone deacetylation, which is thought to lead to a compact chromatin structure to which the accessibility of transcriptional activators is impaired. In a general mechanistic view, the overall picture suggests that transcriptional silencers and co-repressors act in analogy to transcriptional activators and coactivators, but with the opposite effect leading to gene silencing. We provide a comprehensive overview of the currently known higher eukaryotic co-repressors, their mechanism of action, and their involvement in biological and pathophysiological pathways. We also show the different pathways that lead to the regulation of co-repressor-silencer complex formation. PMID- 11023973 TI - Nitrosation and oxidation in the regulation of gene expression. AB - A growing body of evidence suggests that the cellular response to oxidative and nitrosative stress is primarily regulated at the level of transcription. Posttranslational modification of transcription factors may provide a mechanism by which cells sense these redox changes. In bacteria, for example, OxyR senses redox-related changes via oxidation or nitrosylation of a free thiol in the DNA binding region. This mode of regulation may serve as a paradigm for redox-sensing by eukaryotic transcription factors as most-including NF-kappaB, AP-1, and p53 contain reactive thiols in their DNA binding regions, the modification of which alters binding in vitro. Several of these transcription factors have been found to be sensitive to both reactive oxygen species and nitric oxide-related species in vivo. It remains entirely unclear, however, if oxidation or nitrosylation of eukaryotic transcription factors is an important mode of regulation, or whether transcriptional activating pathways are principally controlled at other redox sensitive levels.-Marshall, H. E., Merchant, K., Stamler, J. S. Nitrosation and oxidation in the regulation of gene expression. PMID- 11023974 TI - p53 from complexity to simplicity: mutant p53 stabilization, gain-of-function, and dominant-negative effect. AB - Increasing the complexity of their models, p53s are stabilized either in order to function (wt p53) or due to the loss of function (mutant p53) with acquiring a mysterious prion-like ability to drive the normal p53 into the abnormal conformation to gain new functions. As already recognized, the loss of trans activating function leads to a stabilization of mutant p53 because of the disappearance of the p53-inducible proteins, which otherwise directly (Mdm-2) or indirectly (p21) target p53 for degradation. Simplifying further, I will discuss that the loss of function results in a dominant-negative effect and gain-of function (a dominant-positive effect). Thus, mutant p53 lacking trans-activation function nevertheless may retain the ability to repress transcription due to its competition with numerous transcription factors for their coactivators. When mutant p53 competes with wt p53, the inhibition of the wt p53-dependent transcription is perceived as a dominant-negative effect. Just like trans repression, a dominant-negative effect requires an excess of p53 and, therefore, a 'dominant'-negative effect is not dominant. Furthermore, the stabilization of an endogenous mt p53 due to the loss of wt functions cannot occur in the presence of the wt p53 allele. Given the inability of mutant p53 to accumulate in the presence of wt p53, a dominant-negative effect does not naturally occur and, not surprisingly, heterozygous mt/wt cells are rare. The detection of a dominant negative effect simply indicates that mutant p53 indeed has lost its function. Last, since mutant p53 loses some or most but not all activities and accumulates in the absence of wt allele, gain-of-function can be considered as an exaggeration of the remaining functions. Applications to cancer therapy are discussed. -Blagosklonny, M. V. p53 from complexity to simplicity: mutant p53 stabilization, gain-of-function, and dominant-negative effect. PMID- 11023975 TI - Increased formation and decreased resorption of bone in mice with elevated vitamin D receptor in mature cells of the osteoblastic lineage. AB - The microarchitecture of bone is regulated by complex interactions between the bone-forming and resorbing cells, and several compounds regulate both actions. For example, vitamin D, which is required for bone mineralization, also stimulates bone resorption. Transgenic mice overexpressing the vitamin D receptor solely in mature cells of the osteoblastic bone-forming lineage were generated to test the potential therapeutic value of shifting the balance of vitamin D activity in favor of bone formation. Cortical bone was 5% wider and 15% stronger in these mice due to a doubling of periosteal mineral apposition rate without altered body weight or calcium homeostatic hormone levels. A 20% increase in trabecular bone volume in transgenic vertebrae was also observed, unexpectedly associated with a 30% reduction in resorption surface rather than greater bone formation. These findings indicate anabolic vitamin D activity in bone and identify a previously unknown pathway from mature osteoblastic cells to inhibit osteoclastic bone resorption, counterbalancing the known stimulatory action through immature osteoblastic cells. A therapeutic approach that both stimulates cortical anabolic and inhibits trabecular resorptive pathways would be ideal for treatment of osteoporosis and other osteopenic disorders. PMID- 11023976 TI - Thrombospondin-1/HIV-1 tat protein interaction: modulation of the biological activity of extracellular Tat. AB - Tat protein, a trans-activating factor of the human immunodeficiency virus type 1, acts also as an extracellular molecule modulating gene expression, cell survival, growth, transformation, and angiogenesis. Here we demonstrate that human thrombospondin-1 (TSP), a plasma glycoprotein and constituent of the extracellular matrix, binds to glutathione-S-transferase (GST)-Tat protein but not to GST. Scatchard plot analysis of the binding of free GST-Tat to immobilized TSP reveals a high-affinity interaction (Kd equal to 25 nM). Accordingly, TSP inhibits cell internalization and HIV-1 LTR trans-activating activity of extracellular Tat in HL3T1 cells with ID50 equal to 10-30 nM. Also, TSP inhibits cell interaction and mitogenic activity of extracellular Tat in T53 Tat-less cells. TSP is instead ineffective when administered after the interaction of Tat with cell surface heparan-sulfate proteoglycans has occurred, in keeping with its ability to prevent but not disrupt Tat/heparin interaction in vitro. Finally, TSP inhibits the autocrine loop of stimulation exerted by endogenous Tat in parental T53 cells. Accordingly, TSP overexpression inhibits cell proliferation, angiogenic activity, and tumorigenic capacity of stable T53 transfectants. Our data demonstrate the ability of TSP to bind to Tat protein and to affect its LTR trans-activating, mitogenic, angiogenic, and tumorigenic activity. These findings suggest that TSP may be implicated in the progression of AIDS and in AIDS associated pathologies by modulating the bioavailability and biological activity of extracellular Tat. PMID- 11023977 TI - A role for p75 neurotrophin receptor in the control of apoptosis-driven hair follicle regression. AB - To examine the mechanisms that underlie the neurotrophin-induced, apoptosis driven hair follicle involution (catagen), the expression and function of p75 neurotrophin receptor (p75NTR), which is implicated in apoptosis control, were studied during spontaneous catagen development in murine skin. By RT-PCR, high steady-state p75NTR mRNA skin levels were found during the anagen-catagen transition of the hair follicle. By immunohistochemistry, p75NTR alone was strongly expressed in TUNEL+/Bcl2- keratinocytes of the regressing outer root sheath, but both p75NTR and TrkB and/or TrkC were expressed by the nonregressing TUNEL-/Bcl2+ secondary hair germ keratinocytes. To determine whether p75NTR is functionally involved in catagen control, spontaneous catagen development was compared in vivo between p75NTR knockout (-/-) and wild-type mice. There was significant catagen retardation in p75NTR knockout mice as compared to wild-type controls (P<0.05). Instead, transgenic mice-overexpressing NGF (promoter: K14) showed substantial acceleration of catagen (P<0.001). Although NGF, brain-derived neurotrophic factor (BDNF), and neurotrophin 3 (NT-3) accelerated catagen in the organ-cultured skin of C57BL/6 mice, these neurotrophins failed to promote catagen development in the organ-cultured p75NTR null skin. These findings suggest that p75NTR signaling is involved in the control of kerotinocyte apoptosis during catagen and that pharmacological manipulation of p75NTR signaling may prove useful for the treatment of hair disorders that display premature entry into catagen. PMID- 11023978 TI - ATPase activity of the sulfonylurea receptor: a catalytic function for the KATP channel complex. AB - ATP-sensitive K+ (KATP) channels are unique metabolic sensors formed by association of Kir6.2, an inwardly rectifying K+ channel, and the sulfonylurea receptor SUR, an ATP binding cassette protein. We identified an ATPase activity in immunoprecipitates of cardiac KATP channels and in purified fusion proteins containing nucleotide binding domains NBD1 and NBD2 of the cardiac SUR2A isoform. NBD2 hydrolyzed ATP with a twofold higher rate compared to NBD1. The ATPase required Mg2+ and was insensitive to ouabain, oligomycin, thapsigargin, or levamisole. K1348A and D1469N mutations in NBD2 reduced ATPase activity and produced channels with increased sensitivity to ATP. KATP channel openers, which bind to SUR, promoted ATPase activity in purified sarcolemma. At higher concentrations, openers reduced ATPase activity, possibly through stabilization of MgADP at the channel site. K1348A and D1469N mutations attenuated the effect of openers on KATP channel activity. Opener-induced channel activation was also inhibited by the creatine kinase/creatine phosphate system that removes ADP from the channel complex. Thus, the KATP channel complex functions not only as a K+ conductance, but also as an enzyme regulating nucleotide-dependent channel gating through an intrinsic ATPase activity of the SUR subunit. Modulation of the channel ATPase activity and/or scavenging the product of the ATPase reaction provide novel means to regulate cellular functions associated with KATP channel opening. PMID- 11023979 TI - Hypothermia injury/cold-induced apoptosis--evidence of an increase in chelatable iron causing oxidative injury in spite of low O2-/H2O2 formation. AB - When incubated at 4 degrees C, cultured rat hepatocytes or liver endothelial cells exhibit pronounced injury and, during earlier rewarming, marked apoptosis. Both processes are mediated by reactive oxygen species, and marked protective effects of iron chelators as well as the protection provided by various other antioxidants suggest that hydroxyl radicals, formed by classical Fenton chemistry, are involved. However, when we measured the Fenton chemistry educt hydrogen peroxide and its precursor, the superoxide anion radical, formation of both had markedly decreased and steady-state levels of hydrogen peroxide did not alter during cold incubation of either liver endothelial cells or hepatocytes. Similarly, there was no evidence of an increase in O2-/H2O2 release contributing to cold-induced apoptosis occurring on rewarming. In contrast to the release/level of O2- and H2O2, cellular homeostasis of the transition metal iron is likely to play a key role during cold incubation of cultured hepatocytes: the hepatocellular pool of chelatable iron, measured on a single-cell level using laser scanning microscopy and the fluorescent indicator phen green, increased from 3.1 +/- 2.3 microM (before cold incubation) to 7.7 +/- 2.4 microM within 90 min after initiation of cold incubation. This increase in the cellular chelatable iron pool was reversible on rewarming after short periods of cold incubation. The cold-induced increase in the hepatocellular chelatable iron pool was confirmed using the calcein method. These data suggest that free radical-mediated hypothermia injury/cold-induced apoptosis is primarily evoked by alterations in the cellular iron homeostasis/a rapid increase in the cellular chelatable iron pool and not by increased formation of O2-/H2O2. PMID- 11023980 TI - Increased susceptibility to ischemia-induced brain damage in transgenic mice overexpressing a dominant negative form of SHP2. AB - Cell culture studies have established SH2 domain-containing protein tyrosine phosphatase-2 (SHP2) as an important factor in growth factor and cytokine activated signaling pathways. However, the significance of SHP2 in the mammalian central nervous system (CNS) is not known since early embryonic lethality occurs in shp2 null mice. To bypass this embryonic lethality, transgenic animals containing a catalytically inactive mutant of SHP2 (SHP2-CS) under the control of a nestin intron II/thymidine kinase minimal promoter were generated. In the developing CNS of these animals, although high-level transgene expression was detected in the neuroepithelium, there was no obvious abnormality in progenitor cell proliferation or migration. In the adult brain, high-level transgene expression was detected in the subventricular zone, rostral migratory stream, dentate gyrus of hippocampus, and cerebellum. Because SHP2 function is likely important in cell survival pathways, we used a focal cerebral ischemia model to examined whether SHP2 is important during CNS injury. Ischemia-induced damage and neuronal death was found to be significantly greater in nestin-SHP2-CS mice than in wild-type littermates. These findings indicate that SHP2 is a required factor in signaling pathway(s) important for neuronal survival. PMID- 11023981 TI - Anti-monocyte chemoattractant protein-1 gene therapy inhibits vascular remodeling in rats: blockade of MCP-1 activity after intramuscular transfer of a mutant gene inhibits vascular remodeling induced by chronic blockade of NO synthesis. AB - Monocyte chemoattractant protein-1 (MCP-1) may play an essential part in the formation of arteriosclerosis by recruiting monocytes into the arterial wall. Thus, we devised a new strategy for anti-MCP-1 gene therapy against arteriosclerosis by transfecting an amino-terminal deletion mutant (missing the amino-terminal amino acids 2 to 8) of the human MCP-1 gene into a remote organ (skeletal muscles). Intramuscular transduction with the mutant MCP-1 gene blocked monocyte recruitment induced by a subcutaneous injection of recombinant MCP-1. In a rat model in which the chronic inhibition of endothelial nitric oxide synthesis induces early vascular inflammation as well as subsequent coronary vascular remodeling, this strategy suppressed monocyte recruitment into the coronary vessels and the development of vascular medial thickening, but did not reduce perivascular fibrosis. Thus, MCP-1 is necessary for the development of medial thickening but not for fibrosis in this model. This new strategy may be a useful and feasible gene therapy against arteriosclerosis. PMID- 11023982 TI - Liver regeneration induced by a designer human IL-6/sIL-6R fusion protein reverses severe hepatocellular injury. AB - The cytokine IL-6 plays a significant role in liver regeneration in conjunction with additional growth factors (HGF, TNF-alpha, and TGF-alpha). Many IL-6 effects depend on a naturally occurring soluble IL-6 receptor (sIL-6R). Here, the chimeric protein hyper-IL-6, constructed from the human IL-6 protein fused to a truncated form of its receptor, was found to have superagonistic IL-6 properties, and as such, enhanced liver cell regeneration. Hyper-IL-6 reversed the state of hepatotoxicity and enhanced the survival rates of rats suffering from fulminant hepatic failure after D-galactosamine administration. The hyper-IL-6 protein has a significant potential for use in the treatment of severe human liver diseases. PMID- 11023983 TI - Infusion of recombinant human acid sphingomyelinase into niemann-pick disease mice leads to visceral, but not neurological, correction of the pathophysiology. AB - An inherited deficiency of acid sphingomyelinase (ASM) activity results in the Type A and B forms of Niemann-Pick disease (NPD). Using the ASM-deficient mouse model (ASMKO) of NPD, we evaluated the efficacy of enzyme replacement therapy (ERT) for the treatment of this disorder. Recombinant human ASM (rhASM) was purified from the media of overexpressing Chinese Hamster ovary cells and i.v. injected into 16 five-month-old ASMKO mice at doses of 0.3, 1, 3, or 10 mg/kg every other day for 14 days (7 injections). On day 16, the animals were killed and the tissues were analyzed for their sphingomyelin (SPM) content. Notably, the SPM levels were markedly reduced in the hearts, livers, and spleens of these animals, and to a lesser degree in the lungs. Little or no substrate depletion was found in the kidneys or brains. Based on these results, three additional 5 month-old ASMKO animals were injected every other day with 5 mg/kg for 8 days (4 injections) and killed on day 10 for histological analysis. Consistent with the biochemical results, marked histological improvements were observed in the livers, spleens, and lungs, indicating a reversal of the disease pathology. A group of 10 ASMKO mice were then i.v. injected once a week with 1 mg/kg rhASM for 15 wk, starting at 3 wk of age. Although anti-rhASM antibodies were produced in these mice, the antibodies were not neutralizing and no adverse effects were observed from this treatment. Weight gain and rota-rod performance were slightly improved in the treated animals as compared with ASMKO control animals, but significant neurological deficits were still observed and their life span was not extended by ERT. In contrast with these CNS results, striking histological and biochemical improvements were found in the reticuloendothelial system organs (livers, spleens, and lungs). These studies indicate that ERT should be an effective therapeutic approach for Type B NPD, but is unlikely to prevent the severe neurodegeneration associated with Type A NPD. PMID- 11023984 TI - Mildly oxidized low density lipoprotein activates multiple apoptotic signaling pathways in human coronary cells. AB - Apoptosis of arterial cells induced by oxidized low density lipoproteins (OxLDL) is thought to contribute to the progression of atherosclerosis. However, most data on apoptotic effects and mechanisms of OxLDL were obtained with extensively oxidized LDL unlikely to occur in early stages of atherosclerotic lesions. We now demonstrate that mildly oxidized LDL generated by incubation with oxygen radical producing xanthine/xanthine oxidase (X/XO) induces apoptosis in primary cultures of human coronary endothelial and SMC, as determined by TUNEL technique, DNA laddering, and FACS analysis. Apoptosis was markedly reduced when X/XO-LDL was generated in the presence of different oxygen radical scavengers. Apoptotic signals were mediated by intramembrane domains of both Fas and tumor necrosis factor (TNF) receptors I and II. Blocking of Fas ligand (FasL) reduced apoptosis by 50% and simultaneous blocking of FasL and TNF receptors by 70%. Activation of apoptotic receptors was accompanied by an increase of proapoptotic and a decrease in antiapoptotic proteins of the Bcl-2 family and resulted in marked activation of class I and II caspases. Mildly oxidized LDL also activated MAP and Jun kinases and increased p53 and other transcription factors (ATF-2, ELK-1, CREB, AP 1). Inhibitors of Map and Jun kinase significantly reduced apoptosis. Our results provide the first evidence that OxLDL-induced apoptosis involves TNF receptors and Jun activation. More important, they demonstrate that even mildly oxidized LDL formed in atherosclerotic lesions may activate a broad cascade of oxygen radical-sensitive signaling pathways affecting apoptosis and other processes influencing the evolution of plaques. Thus, we suggest that extensive oxidative modifications of LDL are not necessary to influence signal transduction and transcription in vivo. PMID- 11023985 TI - The Raf-1 kinase associates with vimentin kinases and regulates the structure of vimentin filaments. AB - Using immobilized GST-Raf-1 as bait, we have isolated the intermediate filament protein vimentin as a Raf-1-associated protein. Vimentin coimmunoprecipitated and colocalized with Raf-1 in fibroblasts. Vimentin was not a Raf-1 substrate, but was phosphorylated by Raf-1-associated vimentin kinases. We provide evidence for at least two Raf-1-associated vimentin kinases and identified one as casein kinase 2. They are regulated by Raf-1, since the activation status of Raf-1 correlated with the phosphorylation of vimentin. Vimentin phosphorylation by Raf 1 preparations interfered with its polymerization in vitro. A subset of tryptic vimentin phosphopeptides induced by Raf-1 in vitro matched the vimentin phosphopeptides isolated from v-raf-transfected cells labeled with orthophosphoric acid, indicating that Raf-1 also induces vimentin phosphorylation in intact cells. In NIH 3T3 fibroblasts, the selective activation of an estrogen regulated Raf-1 mutant induced a rearrangement and depolymerization of the reticular vimentin scaffold similar to the changes elicited by serum treatment. The rearrangement of the vimentin network occurred independently of the MEK/ERK pathway. These data identify a new branch point in Raf-1 signaling, which links Raf-1 to changes in the cytoskeletal architecture. PMID- 11023986 TI - Stimulation of protein (collagen) synthesis in sponge cells by a cardiac myotrophin-related molecule from Suberites domuncula. AB - The body wall of sponges (Porifera), the lowest metazoan phylum, is formed by two epithelial cell layers of exopinacocytes and endopinacocytes, both of which are associated with collagen fibrils. Here we show that a myotrophin-like polypeptide from the sponge Suberites domuncula causes the expression of collagen in cells from the same sponge in vitro. The cDNA of the sponge myotrophin was isolated; the potential open reading frame of 360 nt encodes a 120 aa long protein (Mr of 12,837). The sequence SUBDOMYOL shares high similarity with the known metazoan myotrophin sequences. The expression of SUBDOMYOL is low in single cells but high after formation of primmorph aggregates as well as in intact animals. Recombinant myotrophin was found to stimulate protein synthesis by fivefold, as analyzed by incorporation studies using [3H] lysine. In addition, it is shown that after incubation of single cells with myotrophin, the primmorphs show an unusual elongated, oval-shaped appearance. It is demonstrated that in the presence of recombinant myotrophin, the cells up-regulate the expression of the collagen gene. The cDNA for S. domuncula collagen was isolated; the deduced aa sequence shows that the collagenous internal domain is rather short, with only 24 G-x-y collagen triplets. We conclude that the sponge myotrophin causes in homologous cells the same/similar effect as the cardiac myotrophin in mammalian cells, where it is involved in initiation of cardial ventricular hypertrophy. We assume that an understanding of sponge molecular cell biology will also contribute to a further elucidation of human diseases, here of the cardiovascular system. PMID- 11023987 TI - HDL-associated PAF-AH reduces endothelial adhesiveness in apoE-/- mice. AB - Macrophage infiltration into the subendothelial space at lesion prone sites is the primary event in atherogenesis. Inhibition of macrophage homing might therefore prevent atherosclerosis. Since HDL levels are inversely correlated with cardiovascular risk, their effect on macrophage homing was assessed in apoE deficient (apoE-/-) mice. Overexpression of human apolipoprotein AI in apoE-/- mice increased HDL levels 3-fold and reduced macrophage accumulation in an established assay of leukocyte homing to aortic root endothelium 3.2-fold (P<0.005). This was due to reduced in vivo betaVLDL oxidation, reduced betaVLDL triggered endothelial cytosolic Ca2+ signaling through PAF-like bioactivity, lower ICAM-1 and VCAM-1 expression, and diminished ex vivo leukocyte adhesion. Adenoviral gene transfer of human PAF-acetylhydrolase (PAF-AH) in apoE-/- mice increased PAF-AH activity 1.5-fold (P<0.001), reduced betaVLDL-induced ex vivo macrophage adhesion 3.5-fold (P<0.01), and reduced in vivo macrophage homing 2.6 fold (P<0.02). These inhibitory effects were observed in the absence of increased HDL cholesterol levels. In conclusion, HDL reduces macrophage homing to endothelium by reducing oxidative stress via its associated PAF-AH activity. This protective mechanism is independent of the function of HDL as cholesterol acceptor. Modulation of lipoprotein oxidation by PAF-AH may prevent leukocyte recruitment to the vessel wall, a key feature in atherogenesis. PMID- 11023988 TI - The intestinal fatty acid binding protein is not essential for dietary fat absorption in mice. AB - The intestinal fatty acid binding protein (I-FABP) belongs to a family of 15 kDa clamshell-like proteins that are found in many different tissues. So far, nine types have been identified. Their primary structures are highly conserved between species but somewhat less so among the different types. The function of these proteins, many of which are highly expressed, is not well understood. Their ability to bind lipid ligands suggests a role in lipid metabolism, but direct evidence for this idea is still lacking. We tested the hypothesis that I-FABP serves an essential role in the assimilation of dietary fatty acids by disrupting its gene (Fabpi) in the mouse. We discovered that Fabpi-/- mice are viable, but they display alterations in body weight and are hyperinsulinemic. Male Fabpi-/- mice had elevated plasma triacylglycerols and weighed more regardless of the dietary fat content. In contrast, female Fabpi-/- mice gained less weight in response to a high-fat diet. The results clearly demonstrate that I-FABP is not essential for dietary fat absorption. We propose that I-FABP functions as a lipid sensing component of energy homeostasis that alters body weight gain in a gender specific fashion. PMID- 11023989 TI - GD3 ganglioside directly targets mitochondria in a bcl-2-controlled fashion. AB - Lipid and glycolipid diffusible mediators are involved in the intracellular progression and amplification of apoptotic signals. GD3 ganglioside is rapidly synthesized from accumulated ceramide after the clustering of death-inducing receptors and triggers apoptosis. Here we show that GD3 induces dissipation of DeltaPsim and swelling of isolated mitochondria, which results in the mitochondrial release of cytochrome c, apoptosis inducing factor, and caspase 9. Soluble factors released from GD3-treated mitochondria are sufficient to trigger DNA fragmentation in isolated nuclei. All these effects can be blocked by cyclosporin A, suggesting that GD3 is acting at the level of the permeability transition pore complex. We found that endogenous GD3 accumulates within mitochondria of cells undergoing apoptosis after ceramide exposure. Accordingly, suppression of GD3 synthase (ST8) expression in intact cells substantially prevents ceramide-induced DeltaPsim dissipation, indicating that endogenously synthesized GD3 induces mitochondrial changes in vivo. Finally, enforced expression of bcl-2 significantly prevents GD3-induced mitochondrial changes, caspase 9 activation, and apoptosis. These results show that mitochondria are a key destination for apoptogenic GD3 ganglioside along the lipid pathway to programmed cell death and indicate that relevant GD3 targets are under bcl-2 control. PMID- 11023991 TI - Adenosine inhibits IL-12 and TNF-[alpha] production via adenosine A2a receptor dependent and independent mechanisms. AB - Interleukin 12 (IL-12) is a crucial cytokine in the regulation of T helper 1 vs. T helper 2 immune responses. In the present study, we investigated the effect of the endogenous purine nucleoside adenosine on the production of IL-12. In mouse macrophages, adenosine suppressed IL-12 production. Although the order of potency of adenosine receptor agonists suggested the involvement of A2a receptors, data obtained with A2a receptor-deficient mice showed that the adenosine suppression of IL-12 and even TNF-alpha production is only partly mediated by A2a receptor ligation. Studies with adenosine receptor antagonists or the adenosine uptake blocker dipyridamole showed that adenosine released endogenously also decreases IL-12. Although adenosine increases IL-10 production, the inhibition of IL-12 production is independent of the increased IL-10. The mechanism of action of adenosine was not associated with alterations of the activation of the p38 and p42/p44 mitogen-activated protein kinases or the phosphorylation of the c-Jun terminal kinase. Adenosine failed to affect steady-state levels of either IL-12 p35 or p40 mRNA, but augmented IL-10 mRNA levels. In summary, adenosine inhibits IL-12 production via various adenosine receptors. These results support the notion that adenosine-based therapies might be useful in certain autoimmune and/or inflammatory diseases. PMID- 11023990 TI - Differential expression and responsiveness of chemokine receptors (CXCR1-3) by human microvascular endothelial cells and umbilical vein endothelial cells. AB - The basis for the angiogenic effects of CXC chemokines such as interleukin 8 (IL 8) and for angiostatic chemokines such as interferon-inducible protein 10 (IP-10) has been difficult to assess. We recently reported, based on an RNase protection assay, that human umbilical vein endothelial cells (HUVECs) did not express detectable mRNA for the IL-8 receptors CXCR1 and CXCR2. This raised the possibility of heterogeneity of receptor expression by different endothelial cell (ECs) types. Since systemic angiogenesis induced by IL-8 would more likely involve microvessel ECs, we investigated CXC receptor expression on human microvascular dermal endothelial cells (HMECs). By confocal microscopy and immunofluorescence we observed that HMECs consistently expressed high levels of CXCR1 and CXCR4 (mean fluorescence intensity of 261+/-22.1 and 306.2+/-19, respectively) and intermediate levels of CXCR3 and CXCR2 (173.9+/-30. 2 and 156+/ 30.9, respectively). In contrast, only a small proportion of HUVEC preparations expressed low levels of CXCR1, -2, and -3 (66+/-19.9; 49+/-15, and 81.4+/-17.9, respectively). However, both HMECs and HUVECs expressed equal levels of CXCR4. As expected, HMECs had more potent chemotactic responses to IL-8 than HUVECs, and this was correlated with the levels of IL-8 receptors on the ECs. Antibodies to CXCR1 and CXCR2 each had inhibitory effects on chemotaxis of HMECs to IL-8, indicating that both IL-8 receptors contributed to the migratory response of these cells toward IL-8. Assessment of the functional capacity of CXCR3 unexpectedly revealed that HMECs migrated in response to relatively higher concentrations (100-500 ng/ml) of each of the 'angiostatic' chemokines IP-10, ITAC, and MIG. Despite this, the 'angiostatic' chemokines inhibited the chemotactic response of HMECs to IL-8. IL-8 and SDF-1alpha but not IP-10 induced calcium mobilization in adherent ECs, suggesting that signaling events associated with calcium mobilization are separable from those required for chemotaxis. Taken together, our data indicated that functional differences among EC types is dependent on the level of the expression of CXC chemokine receptors. Whether this heterogeneity in receptor expression by ECs reflects distinct differentiation pathways remains to be established. PMID- 11023992 TI - Cathepsin L deficiency as molecular defect of furless: hyperproliferation of keratinocytes and pertubation of hair follicle cycling. AB - Lysosomal cysteine proteinases of the papain family are involved in lysosomal bulk proteolysis, major histocompatibility complex class II mediated antigen presentation, prohormone processing, and extracellular matrix remodeling. Cathepsin L (CTSL) is a ubiquitously expressed major representative of the papain like family of cysteine proteinases. To investigate CTSL in vivo functions, the gene was inactivated by gene targeting in embryonic stem cells. CTSL-deficient mice develop periodic hair loss and epidermal hyperplasia, acanthosis, and hyperkeratosis. The hair loss is due to alterations of hair follicle morphogenesis and cycling, dilatation of hair follicle canals, and disturbed club hair formation. Hyperproliferation of hair follicle epithelial cells and basal epidermal keratinocytes-both of ectodermal origin-are the primary characteristics underlying the mutant phenotype. Pathological inflammatory responses have been excluded as a putative cause of the skin and hair disorder. The phenotype of CTSL deficient mice is reminiscent of the spontaneous mouse mutant furless (fs). Analyses of the ctsl gene of fs mice revealed a G149R mutation inactivating the proteinase activity. CTSL is the first lysosomal proteinase shown to be essential for epidermal homeostasis and regular hair follicle morphogenesis and cycling. PMID- 11023993 TI - VEGF-C and VEGF-D expression in neuroendocrine cells and their receptor, VEGFR-3, in fenestrated blood vessels in human tissues. AB - Recently, vascular endothelial growth factor receptor 3 (VEGFR-3) has been shown to provide a specific marker for lymphatic endothelia in certain human tissues. In this study, we have investigated the expression of VEGFR-3 and its ligands VEGF-C and VEGF-D in fetal and adult tissues. VEGFR-3 was consistently detected in the endothelium of lymphatic vessels such as the thoracic duct, but fenestrated capillaries of several organs including the bone marrow, splenic and hepatic sinusoids, kidney glomeruli and endocrine glands also expressed this receptor. VEGF-C and VEGF-D, which bind both VEGFR-2 and VEGFR-3 were expressed in vascular smooth muscle cells. In addition, intense cytoplasmic staining for VEGF-C was observed in neuroendocrine cells such as the alpha cells of the islets of Langerhans, prolactin secreting cells of the anterior pituitary, adrenal medullary cells, and dispersed neuroendocrine cells of the gastrointestinal tract. VEGF-D was observed in the innermost zone of the adrenal cortex and in certain dispersed neuroendocrine cells. These results suggest that VEGF-C and VEGF-D have a paracrine function and perhaps a role in peptide release from secretory granules of certain neuroendocrine cells to surrounding capillaries. PMID- 11023994 TI - The loss of sympathetic nerve fibers in the synovial tissue of patients with rheumatoid arthritis is accompanied by increased norepinephrine release from synovial macrophages. AB - Our objective was to investigate sympathetic and sensory nerve fibers in synovial tissue in rheumatoid arthritis (RA) and osteoarthritis (OA) in relation to histological inflammation and synovial cytokine and norepinephrine (NE) secretion. Immunohistochemistry was used to detect nerve fibers and inflammatory parameters. A superfusion technique of synovial tissue pieces was used to investigate cytokine and NE secretion. In RA, we detected 0.2 +/- 0.04 tyrosine hydroxylase-positive (TH-positive=sympathetic) nerve fibers/mm2 as compared to 4.4 +/- 0. 8 nerve fibers/mm2 in OA (P<0.001). In RA, there was a negative correlation between the number of TH-positive nerve fibers and inflammation index (RRank=-0.705, P=0.002) and synovial IL-6 secretion (RRank=-0.630, P=0.009), which was not found in OA. Substance P-positive (=sensory) nerve fibers were increased in RA as compared to OA (3.5+/-0.2 vs. 2.3+/-0.3/mm2, P=0.009). Despite lower numbers of sympathetic nerve fibers in RA than in OA, NE release was similar at baseline (RA vs. OA: 152+/-36 vs. 106+/-21 pg/ml, n.s.). Basal synovial NE secretions correlate with the number of TH-positive CD 163+ synovial macrophages (RA: RRank=0.622, P=0.031; OA: RRank=0.299, n.s.), and synovial macrophages have been shown to produce NE in vitro. Whereas sympathetic innervation is reduced, sensory innervation is increased in the synovium from patients with longstanding RA when compared to the synovium from OA patients. The differential patterns of innervation are dependent on the severity of the inflammation. However, NE secretion from the synovial tissue is maintained by synovial macrophages. This demonstrates a loss of the influence of the sympathetic nervous system on the inflammation, accompanied by an up-regulation of the sensory inputs into the joint, which may contribute to the maintenance of the disease. PMID- 11023995 TI - In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate. AB - To target disseminated tumors in vivo, transgenes [beta-galactosidase gene, green fluorescence protein (GFP) gene, herpes simplex virus thymidine kinase (HSV-TK)] were conjugated to transferrin (Tf) by a biotin-streptavidin bridging, which is stoichiometrically controllable, and Tf receptor (Tf-R) affinity chromatography, which selects Tf conjugates with intact receptor bindings sites from reacting with the linker. Tf-beta-galactosidase plasmid conjugate thus constructed was specifically transfected to human erythroleukemia cells (K562) via Tf-R without the aid of any lysosomotropic agents. The transfection efficiency of the conjugate was superior to those of lipofection (1% staining) and retroviral vector (5%) and slightly lower than that of adenovirus (70%). The high level of expression with our conjugate was confirmed using other tumor cells (M7609, TMK 1) whereas in normal diploid cells (HEL), which express low levels of Tf-R, expression was negligible. When GFP gene conjugates were systemically administered through the tail vein to nude mice subcutaneously inoculated with tumor, expression of GFP mRNA was found almost exclusively in tumors and to a much lesser extent in muscles, whereas GFP revealed by fluorescence microscopy was detected only in the former. To exploit a therapeutic applicability of this method, suicide gene therapy using Tf-HSV-TK gene conjugate for massively metastasized k562 tumors in severe combined immune-deficient mice was conducted, and a marked prolongation of survival and significant reduction of tumor burden were confirmed. Thus, this method could also be used for gene therapy to disseminated tumors. PMID- 11023997 TI - Erratum PMID- 11023996 TI - Regulation of CYP26 (cytochrome P450RAI) mRNA expression and retinoic acid metabolism by retinoids and dietary vitamin A in liver of mice and rats. AB - Retinoic acid (RA), through nuclear retinoid receptors, regulates the expression of numerous genes. However, little is known of the biochemical mechanisms that regulate RA concentration in vivo. CYP26 (P450RAI), a novel cytochrome P450, is expressed during embryonic development, induced by all-trans RA, and capable of catalyzing the oxidation of [3H]RA to polar retinoids including 4-oxo-RA. Here we report that CYP26 expression in adult liver is regulated by all-trans RA and dietary vitamin A, and is correlated with the metabolism of all-trans RA to polar metabolites. In normal mouse and rat liver, CYP26 mRNA was barely detectable; however, after acute treatment with all-trans RA CYP26 mRNA and RA metabolism by liver microsomes were significantly induced. Aqueous-soluble RA metabolites were detected, but their formation was not induced. The expression of retinoid receptors, RAR-gamma and RXR-alpha, was not changed after RA treatment in vivo. In a model of chronic vitamin A ingestion during aging, CYP26 mRNA expression, determined by Northern blot and RT-PCR analysis, increased progressively with dietary vitamin A (P<0.0001; marginal < control < supplemented) and age (P<0.003). The relative expression of CYP26 mRNA was positively correlated with liver total retinol (log10), ranging from undetectable CYP26 expression at liver retinol concentrations below approximately 20 nmol/g to a three- to fourfold elevation at concentrations >10,000 nmol/g (r=0.90, P<0.0001). We conclude that CYP26 expression and RA metabolism are regulated in adult liver not only acutely by RA administration, as may be relevant to retinoid therapy, but under chronic dietary conditions relevant to vitamin A nutrition in humans. PMID- 11023998 TI - Preconditioning regulation of bcl-2 and p66shc by human NOS1 enhances tolerance to oxidative stress. AB - Preconditioning stress induced by a transient ischemia may increase brain tolerance to oxidative stress, and the underlying neuroprotective mechanisms are not well understood. In a series of experiments, we found that endogenous nitric oxide (NO), S-nitrosoglutathione (GSNO), and antioxidants blocked serum deprivation-induced oxidative stress and apoptosis in human neuroblastoma cells. Similar to nuclear redox factor-1 (Ref-1), mRNA of human neuronal nitric oxide synthase (hNOS1) was maximally up-regulated within 2 h after oxidative stress and down-regulated by NO/GSNO and hydroxyl radical (OH) scavenger. A brief preconditioning stress induced by serum deprivation for 2 h caused a delayed increase in the expression of hNOS1 protein and the associated formation of NO and cGMP, which in turn decreased OH generation and stress-related cell death. In addition to inhibiting caspase-3 through a dithiothreitol-sensitive S nitrosylation process, preconditioning stress concomitantly up-regulated the expression of the anti-apoptotic bcl-2 protein and down-regulated the p66shc adaptor protein. This beneficial cytoprotective process of preconditioning stress is mediated by newly synthesized NO because it can be suppressed by the inhibition of hNOS1 and guanylyl cyclase. Therefore, the constitutive isoform of hNOS1 is dynamically redox-regulated to meet both functional and compensatory demands of NO for gene regulation, antioxidant defense, and tolerance to oxidative stress. PMID- 11023999 TI - The Werner syndrome protein contributes to induction of p53 by DNA damage. AB - Mutations in the p53 tumor-suppressor gene promote increased genomic instability and cancer. Mutations in the WRN gene, encoding a DNA helicase, underlie the segmental progeroid Werner syndrome (WS). WS is also associated with increased genomic instability and elevated cancer risk. The p53 and WRN proteins can engage in direct protein-protein interactions. We report that excess WRN elicits increased cellular p53 levels and potentiates p53-mediated apoptosis. Importantly, cells derived from WS patients exhibit an attenuated and delayed induction of p53 by UV or by the topoisomerase I inhibitor camptothecin. These results suggest that WRN may participate in the activation of p53 in response to certain types of DNA damage. Furthermore, the failure to induce p53 effectively may contribute to enhanced genomic instability and elevated cancer risk in WS patients. PMID- 11024001 TI - The human uncoupling protein-3 gene promoter requires MyoD and is induced by retinoic acid in muscle cells. AB - The uncoupling protein-3 (UCP-3) gene encodes for a mitochondrial protein expressed preferentially in skeletal muscle. UCP-3 mRNA is expressed in cultured muscle cells (C2C12 or L6E9) only when differentiated, at which stage UCP-3 is highly induced by all-trans retinoic acid (RA). Here we report that human UCP-3 promoter activity is dependent on MyoD and inducible by all trans-RA. The action of all trans-RA is increased by co-transfection with RA receptor (RAR). We have characterized the RA response element that controls the induction by RA in the 5' noncoding region of the UCP-3 gene. Deletion and point-mutation analysis of the hUCP-3 promoter led us to identify a direct-repeat element with one base-pair spacing (DR1) at position -71/-59 responsible for the induction by RA of the activity of the promoter. This DR1 element bound a nuclear protein complex from muscle cells that contain RAR and retinoid X receptor (RXR). In the absence of this element, the promoter became unresponsive to RA, but it was still dependent on MyoD. In conclusion, it has been established that UCP-3 gene promoter activity is dependent on MyoD, and the first regulatory pathway for UCP-3 gene promoter regulation has been recognized by identifying RA as a transcriptional activator of the gene. PMID- 11024000 TI - An intervention resembling caloric restriction prolongs life span and retards aging in yeast. AB - The yeast Saccharomyces cerevisiae has a finite life span that is measured by the number of daughter cells an individual produces. The 20 genes known to determine yeast life span appear to function in more than one pathway, implicating a variety of physiological processes in yeast longevity. Less attention has been focused on environmental effects on yeast aging. We have examined the role that nutritional status plays in determining yeast life span. Reduction of the glucose concentration in the medium led to an increase in life span and to a delay in appearance of an aging phenotype. The increase in life span was the more extensive the lower the glucose levels. Life extension was also elicited by decreasing the amino acids content of the medium. This suggests that it is the decline in calories and not a particular nutrient that is responsible, in striking similarity to the effect on aging of caloric restriction in mammals. The caloric restriction effect did not require the induction of the retrograde response pathway, which signals the functional status of the mitochondrion and determines longevity. Furthermore, deletion of RTG3, a downstream mediator in this pathway, and caloric restriction had an additive effect, resulting in the largest increase (123%) in longevity described thus far in yeast. Thus, retrograde response and caloric restriction operate along distinct pathways in determining yeast longevity. These pathways may be exclusive, at least in part. This provides evidence for multiple mechanisms of metabolic control in yeast aging. Inasmuch as caloric restriction lowers blood glucose levels, this study raises the possibility that reduced glucose alters aging at the cellular level in mammals. PMID- 11024002 TI - New agents for the treatment of infarcted myocardium. AB - Local delivery of angiogenic growth factors for the treatment of myocardial ischemia has been well documented in various animal models, and clinical trials are now in progress. Our strategy was radically different, based on selective protection of some of the growth factors naturally present within the injured tissue. This protection was obtained by applying a chemically defined substitute for Dextran called RGTA11 (for ReGeneraTing Agent). RGTA is a family of agents, which has properties mimicking those of heparan sulfates toward heparin-binding growth factors (HBGF) and which stimulate tissue repair and protection. Indeed, we have previously shown that RGTA prevents most of the damage resulting from acute skeletal muscle ischemia [FASEB J. (1999) 13, 761-766]. We now show that the same agent can be used for the treatment of myocardial infarction. Acute myocardial infarction was induced in pigs by ligation of the left circumflex artery. One hour later, a single injection of 10 mg of RGTA11 was made in the center of the infarcted area. Three weeks later we observed 1) recovery of 84% of the initial left ventricular ejection fraction (only 55% in saline-treated controls), 2) an almost 50% reduction in the infarct size, 3) a reduction in fibrotic tissue formation, 4) significant preservation of myocytes, and 5) an increase in the number of blood vessels. The treatment of ischemic heart disease with RGTA would have clear advantages over other therapies such as growth factor, gene, or cell transplants, based on a stable, simple, and easy-to-develop chemical product. PMID- 11024003 TI - Stress-mediated inhibition of the classical nuclear protein import pathway and nuclear accumulation of the small GTPase Gsp1p. AB - Stress modifies all aspects of cellular physiology, including the targeting of macromolecules to the nucleus. To determine how distinct types of stress affect classical nuclear protein import, we followed the distribution of NLS-GFP, a reporter protein containing a classical nuclear localization sequence (NLS) fused to green fluorescent protein GFP. Nuclear accumulation of NLS-GFP requires import to be constitutively active; inhibition of import redistributes NLS-GFP throughout the nucleus and cytoplasm. In the yeast Saccharomyces cerevisiae, starvation, heat shock, ethanol and hydrogen peroxide rapidly inhibited classical nuclear import, whereas osmotic stress had no effect. To define the mechanisms underlying the inhibition of classical nuclear import, we located soluble components of the nuclear transport apparatus. Failure to accumulate NLS-GFP in the nucleus always correlated with a redistribution of the small GTPase Gsp1p. Whereas predominantly nuclear under normal conditions, Gsp1p equilibrated between nucleus and cytoplasm in cells exposed to starvation, heat, ethanol or hydrogen peroxide. Furthermore, analysis of yeast strains carrying mutations in different nuclear transport factors demonstrated a role for NTF2, PRP20 and MOG1 in establishing a Gsp1p gradient, as conditional lethal alleles of NTF2 and PRP20 or a deletion of MOG1 prevented Gsp1p nuclear accumulation. On the basis of these results, we now propose that certain types of stress release Gsp1p from its nuclear anchors, thereby promoting a collapse of the nucleocytoplasmic Gsp1p gradient and inhibiting classical nuclear protein import. PMID- 11024004 TI - Cell-free assays for gamma-secretase activity. AB - The amyloid b-protein (Ab) deposited in Alzheimer's disease (AD) is a normally secreted proteolytic product of the amyloid b-protein precursor (APP). Generation of Ab from the APP requires two sequential proteolytic events: an initial b secretase cleavage at the amino terminus of the Ab sequence followed by g secretase cleavage at the carboxyl terminus of Ab. We describe the development of a robust in vitro assay for g-secretase cleavage by showing de novo Ab production in vitro and establish that this assay monitors authentic gamma-secretase activity by documenting the production of a cognate g-CTF, confirming the size of the Ab produced by mass spectrometry, and inhibiting cleavage in this system with multiple inhibitors that alter g-secretase activity in living cells. Using this assay, we demonstrate that the g-secretase activity 1) is tightly associated with the membrane, 2) can be solubilized, 3) has a pH optimum of 6.8 but is active from pH 6.0 to pH >8.4, and 4) ascertain that activities of the g-40 and g-42 are indeed pharmacologically distinct. These studies should facilitate the purification of the protease or proteases that are responsible for this unusual activity, which is a major therapeutic target for the treatment of AD. PMID- 11024005 TI - Advanced glycation end-product receptor interactions on microvascular cells occur within caveolin-rich membrane domains. AB - Advanced glycation end products (AGEs) have an important role in diabetic complications, with many responses mediated through AGE-receptors. The current study has investigated the binding and uptake of AGEs by retinal microvascular endothelium in an attempt to understand the nature of AGE-interaction with receptors on the cell surface. There has been special emphasis placed on the R1, R2, and R3 components of AGE-receptor complex (AGE-RC) and their localization to caveolin-rich membrane domains. Retinal microvascular endothelial cells (RMECs) were exposed to either AGE-modified BSA (AGE-BSA) or native BSA conjugated to colloidal gold (gAGE, gBSA) for various time periods, fixed, and processed for transmission electron microscopy (TEM). Localization of AGE-RC components in caveolae was investigated using confocal microscopy and ultrastructural immunogold labeling. Caveolae were extracted from RMECs using differential Triton X-100 solubility, and Western analysis was conducted to test for caveolae enrichment and the presence of AGE-RC complex components. Ligand blots determined 125I-AGE-BSA binding to caveolae-enriched extracts. Colloidal gold conjugates of AGE-BSA bound to caveolae and were internalized to be trafficked to lysosomal like compartments. AGE-receptor complex components were significantly enriched within caveolae. The data suggest that AGEs interact with their receptors within caveolae. It is significant that the AGE-R complex localizes to these organelles, because this may have implications for AGE binding, internalization, signal transduction, and the modulation of AGE-receptor-mediated vascular cell dysfunction. PMID- 11024006 TI - Anti-inflammatory effects of sodium butyrate on human monocytes: potent inhibition of IL-12 and up-regulation of IL-10 production. AB - Cytokines are critical in regulating unresponsiveness versus immunity towards enteric antigens derived from the intestinal flora and ingested food. There is increasing evidence that butyrate, a major metabolite of intestinal bacteria and crucial energy source for gut epithelial cells, also possesses anti-inflammatory properties. Its influence on cytokine production, however, is not established. Here, we report that butyrate strongly inhibits interleukin-12 (IL-12) production by suppression of both IL-12p35 and IL-12p40 mRNA accumulation, but massively enhances IL-10 secretion in Staphylococcus aureus cell-stimulated human monocytes. The effect of butyrate on IL-12 production was irreversible upon the addition of neutralizing antibodies to IL-10 or transforming growth factor b1 and of indomethacin. In anti-CD3-stimulated peripheral blood mononuclear cells, butyrate enhanced IL-10 and IL-4 secretion but reduced the release of IL-2 and interferon-g. The latter effect was in part a result of suppressed IL-12 production but also a result of inhibition of IL-12 receptor expression on T cells. These data demonstrate a novel anti-inflammatory property of butyrate that may have broad implications for the regulation of immune responses in vivo and could be exploited as new therapeutic approach in inflammatory conditions. PMID- 11024007 TI - alpha-Tocopherol and protein kinase C inhibition enhance platelet-derived nitric oxide release. AB - Platelet activation is tightly regulated by products of the endothelium and platelets including nitric oxide (NO). Excess vascular oxidative stress has been associated with impaired NO release, and antioxidant status has been shown to alter endothelium-derived NO bioactivity. Although physiological levels of a tocopherol are known to inhibit platelet function, the effect of a-tocopherol on platelet NO release is unknown. Loading platelets with physiologic levels of a tocopherol increased platelet NO production approximately 1.5-fold (Pa tocopherol, platelet NO release increased 50% (Pa-Tocopherol-loaded platelets also produced 74% less superoxide as compared with control (Pa-tocopherol inhibited PKC-dependent eNOS phosphorylation as determined by immunoprecipitation. Lastly, platelets isolated from NOS3-deficient mice released 80% less superoxide as compared with control animals (P=0.011), and incubation of NOS III-deficient platelets with 500 mM a-tocopherol only caused a modest additional decrease in platelet superoxide release (NS). Thus, a-tocopherol appears to enhance platelet NO release both in vitro and in vivo through antioxidant- and PKC-dependent mechanisms. PMID- 11024008 TI - The terminal complement complex C5b-9 stimulates interleukin-6 production in human smooth muscle cells through activation of transcription factors NF-kappa B and AP-1. AB - Activation of the complement system plays an important role in the pathogenesis of atherosclerosis. The proinflammatory cytokine interleukin (IL)-6 is potentially involved in the progression of the disease. We therefore investigated whether the terminal complement complex C5b-9 affects IL-6 production from vascular smooth-muscle cells (VSMC) and set out to determine the underlying signal transduction pathway. Stimulation of human VSMC with C5b-9 resulted in an increase of IL-6 transcript and production of IL-6 protein. Pretreatment with pertussis toxin or pyrrolidine dithiocarbamate inhibited complement-dependent IL 6 mRNA expression and IL-6 release, suggesting the involvement of Gi-proteins and nuclear factor-kB (NF-kB). C5b-9 also induced formation of reactive oxygen species, which, along with IL-6 release, was inhibited by the antioxidant N acetylcysteine. C5b-9 activated the redox-sensitive transcription factors NF-kB and activator protein-1 (AP-1), which were both involved in the induction of IL-6 by C5b-9, as demonstrated by cis element double-stranded (decoy) oligonucleotides (ODN). The results demonstrate that activation of the complement system induces IL-6 release from human VSMC by a Gi-dependent pathway involving the generation of oxidative stressand the activation of the redox sensitive transcription factors NF-kB and AP-1. Our data support a new mechanism for the proatherogenic effect of the terminal complement complex. PMID- 11024009 TI - The angiogenesis inhibitor endostatin impairs blood vessel maturation during wound healing. AB - Endostatin is a cleavage product of collagen XVIII that strongly inhibits tumor angiogenesis. To determine if endostatin affects other angiogenic processes, we generated full-thickness excisional wounds on the back of mice that were systemically treated with recombinant murine endostatin. No macroscopic abnormalities of the wound healing process were observed. Histological analysis revealed normal wound contraction and re-epithelialization, but a slight reduction in granulation tissue formation and reduced matrix deposition at the wound edge. The blood vessel density in the wounds of endostatin-treated mice was not affected. However, ultrastructural analysis demonstrated severe abnormalities in blood vessel maturation. The wound vessels in the endostatin-treated mice were narrowed or closed with an irregular luminal surface, resulting in a severe reduction in the number of functional vessels and extravasation of erythrocytes. Endostatin treatment did not affect the expression level and localization of collagen XVIII mRNA and protein. Furthermore, the angiogenesis regulators vascular endothelial growth factor, angiopoietin-1, and angiopoietin-2 were normally expressed in the wounds of endostatin-treated mice. However, expression of the major wound matrix proteins fibronectin and collagens I and III was significantly reduced. This reduction is likely to explain the reduced density of the wound matrix. Our results demonstrate that endostatin treatment reduces the number of functional blood vessels and the matrix density in the granulation tissue, but does not significantly affect the overall wound healing process. PMID- 11024010 TI - Altered expression and functional profile of lysophosphatidic acid receptors in mitogen-activated human blood T lymphocytes. AB - Lysophosphatidic acid (LPA) from platelets and mononuclear phagocytes regulates T cell functions through endothelial differentiation gene-encoded G protein-coupled receptors (Edg Rs). Human blood unactivated CD4+ T cells express predominant ly Edg-4 LPA R over marginal levels of Edg-2 LPA R, as assessed by semiquantitative PCR and Western blots. After mitogen activation, the CD4+ T cells express Ed g-2 Rs at approximately one half the level of Edg-4 Rs. Secretion of IL-2 by unactivated Edg-4 R-predominant CD4+ T cells incubated with anti-CD3 plus anti CD28 antibodies was suppressed significantly and by up to 60% by 10-10 M to 10-6 M LPA, whereas secretion of IL-2 by mitogen-activated Edg-2 R and Edg-4 R codominant CD4+ T cells was enhanced by up to twofold by the same concentrations of LPA. The possibility that the two Edg Rs transduce different LPA signals to CD4+ T cells was supported by findings that IL-2 secretion was inhibited by mouse anti-Edg-4 R monoclonal antibody, but enhanced by mouse anti-Edg-2 R monoclonal antibody. The separate effects of each LPA R were studied in Jurkat T cell transfectants expressing principally human Edg-2 Rs (Jurkat-T-2) or Edg-4 Rs (Jurkat-T-4) and stimulated with anti-CD3 plus phorbol myristate acetate. LPA and anti-Edg-4 R antibody suppressed IL-2 secretion by stimulated Jurkat-T-4 cells, whereas LPA and anti-Edg-2 R antibody enhanced IL-2 secretion by stimulated Jurkat-T-2 cells. Activation-induced alterations in the relative levels of Edg-2 and -4 Rs on CD4+ T cells thus reverse the effects of LPA on T cell receptor stimulated generation of IL-2. PMID- 11024011 TI - Heat shock enhances transcriptional activation of the murine-inducible nitric oxide synthase gene. AB - There is considerable interest in determining the conditions leading to enhanced inducible nitric oxide synthase (iNOS) gene expression and nitric oxide (NO) biosynthesis. Using in vivo footprinting, we demonstrate that heat shock of murine macrophages concurrent with lipopolysaccharide (LPS) treatment stimulated changes in guanine methylation sensitivity at ?898/9, at a putative partial heat shock element (HSE) and at -893/4, a site bordering an E-box, within the iNOS gene enhancer, suggesting inducible occupation by transcription factors at these regions. LPS treatment accompanied by heat shock provoked increased iNOS gene transcription, increased levels of iNOS protein, and increased production of NO compared with LPS treatment alone. Electrophoretic mobility shift analysis revealed low constitutive levels of specific binding to an E-box and a partial HSE within the iNOS enhancer. Binding to the E-box was increased by LPS treatment or by heat shock, achieving a greater increase by a combination of both treatments. The proteins occupying this site were identified as belonging to the USF family of transcription factors. Heat shock or LPS increased binding to the HSE, and the factor responsible for this interaction was identified as heeat shock factor-1 (HSF-1). Mutations at the HSE revealed the importance of HSF-1 in the induction of iNOS by LPS. Thus, our data reveal two novel regulatory sites in the murine iNOS gene, one of which is implicated in enhancing iNOS expression via LPS stimulation, and provide the first evidence that heat shock enhances transcription of the iNOS gene. These results could have implications in the host response mechanism to fever-associated gram-negative infection. PMID- 11024012 TI - Topological organization of the hyaluronan synthase from Streptococcus pyogenes. AB - Since we first reported (DeAngelis, P. L., Papaconstantinou, J., and Weigel, P. H. (1993) J. Biol. Chem. 268, 19181-19184) the cloning of the hyaluronan (HA) synthase from Streptococcus pyogenes (spHAS), numerous membrane-bound HA synthases have been discovered in both prokaryotes and eukaryotes. The HASs are unique among enzymes studied to date because they mediate 6-7 discrete functions in order to assemble a polysaccharide containing hetero-disaccharide units and simultaneously effect translocation of the growing HA chain through the plasma membrane. To understand how the relatively small spHAS performs these various functions, we investigated the topological organization of the protein utilizing fusion analysis with two reporter enzymes, alkaline phosphatase and beta galactosidase, as well as several other approaches. From these studies, we conclude that the NH2 terminus and the COOH terminus, as well as the major portion of a large central domain are localized intracellularly. The first two predicted membrane domains were confirmed to be transmembrane domains and give rise to a very small extracellular loop that is inaccessible to proteases. Several regions of the large internal central domain appear to be associated with, but do not traverse, the membrane. Following the central domain, there are two additional transmembrane domains connected by a second small extracellular loop that also is inaccessible to proteases. The COOH-terminal approximately 25% of spHAS also contains a membrane domain that does not traverse the membrane and may contain extensive re-entrant loops or amphipathic helices. Numerous membrane associations of this latter COOH-terminal region and the central domain may be required to create a pore-like structure through which a growing HA chain can be extruded to the cell exterior. Based on the high degree of similarity among Class I HAS family members, these enzymes may have a similar topological organization for their spHAS-related domains. PMID- 11024013 TI - In vivo regulation of syndecan-3 expression in the rat uterus by 17 beta estradiol. AB - The immature rat uterus has been extensively used as an in vivo model system to study the molecular mechanisms of steroid hormone actions. In this study, we demonstrated the regulated expression of syndecan-3 in the rat uterus by the steroid hormone 17 beta-estradiol. Administration of a single physiological dose of 17 beta-estradiol (40 microg/kg) to ovariectomized immature animals induced a rapid and transient increase in uterine syndecan-3 mRNA. Transcript levels reached a peak elevation of 3-fold above saline control tissues 4 h after hormone administration. Inhibition of message up-regulation by actinomycin D but not cycloheximide indicated a hormone response dependent on RNA transcription but not new protein synthesis. The estrogenic ligands estriol and tamoxifen were also effective at raising syndecan-3 mRNA levels; however, nonestrogenic ligands, including progesterone, 5 alpha-dihydrotestosterone, and dexamethasone, failed to stimulate a change in mRNA levels. Hormone-induced changes in mRNA led to transient changes in syndecan-3 protein content and significant alteration in the temporal and spatial expression in endometrial epithelial cells. Collectively, these data show that the steroid hormone 17 beta-estradiol, regulates transcription of the syndecan-3 gene in the uterus via an estrogen receptor dependent mechanism. This estrogen-regulated expression of syndecan-3 may play an important role in changes in tissue ultrastructure crucial for proper uterine growth. PMID- 11024014 TI - Interaction of MyoD family proteins with enhancers of acetylcholine receptor subunit genes in vivo. AB - The myogenic determination factors (MDFs) are transcriptional activators that target E boxes in many muscle-specific promoters, including those of the genes coding for the subunits of the acetylcholine receptor. It is not known, however, if in vivo a given E box in a transcriptionally active gene is occupied, either uniquely by one MDF or randomly by all MDFs. We have analyzed expression of MDF and acetylcholine receptor subunits in cultured mouse muscle cells and, using chromatin immunoprecipitation, have determined which individual MDFs reside at promoters of several receptor subunit genes. We find that before fusion, C2C12 cells express myf-5, MyoD, and myogenin, all of which take up residence at promoters of all subunits except epsilon. At this stage, herculin is present in limited amounts and is detected mainly at the gamma and delta subunit genes. On myotube formation, herculin reaches high levels; concomitantly, the epsilon subunit gene becomes a common MDF target and begins to be expressed. In general, any MDF protein that is expressed also is present on transcriptionally active receptor genes; transcriptional activity of target genes correlates with occupancy by MDF, in particular, herculin. PMID- 11024015 TI - Identification and characterization of two G protein-coupled receptors for neuropeptide FF. AB - The central nervous system octapeptide, neuropeptide FF (NPFF), is believed to play a role in pain modulation and opiate tolerance. Two G protein-coupled receptors, NPFF1 and NPFF2, were isolated from human and rat central nervous system tissues. NPFF specifically bound to NPFF1 (K(d) = 1.13 nm) and NPFF2 (K(d) = 0.37 nm), and both receptors were activated by NPFF in a variety of heterologous expression systems. The localization of mRNA and binding sites of these receptors in the dorsal horn of the spinal cord, the lateral hypothalamus, the spinal trigeminal nuclei, and the thalamic nuclei supports a role for NPFF in pain modulation. Among the receptors with the highest amino acid sequence homology to NPFF1 and NPFF2 are members of the orexin, NPY, and cholecystokinin families, which have been implicated in feeding. These similarities together with the finding that BIBP3226, an anorexigenic Y1 receptor ligand, also binds to NPFF1 suggest a potential role for NPFF1 in feeding. The identification of NPFF1 and NPFF2 will help delineate their roles in these and other physiological functions. PMID- 11024016 TI - Purification and characterization of pol kappa, a DNA polymerase encoded by the human DINB1 gene. AB - The Escherichia coli dinB gene encodes DNA polymerase (pol) IV, a protein involved in increasing spontaneous mutations in vivo. The protein-coding region of DINB1, the human ortholog of DNA pol IV, was fused to glutathione S transferase and expressed in insect cells. The purified fusion protein was shown to be a template-directed DNA polymerase that we propose to designate pol kappa. Human pol kappa lacks detectable 3' --> 5' proofreading exonuclease activity and is not stimulated by recombinant human proliferating cell nuclear antigen in vitro. Between pH 6.5 and 8.5, human pol kappa possesses optimal activity at 37 degrees C over the pH range 6.5-7.5, and is insensitive to inhibition by aphidicolin, dideoxynucleotides, or NaCl up to 50 mm. Either Mg(2+) or Mn(2+) can satisfy a metal cofactor requirement for pol kappa activity, with Mg(2+) being preferred. Human pol kappa is unable to bypass a cisplatin adduct in the template. However, pol kappa shows limited bypass of an 2-acetylaminofluorene lesion and can incorporate dCTP or dTTP across from this lesion, suggesting that the bypass is potentially mutagenic. These results are consistent with a model in which pol kappa acts as a specialized DNA polymerase whose possible role is to facilitate the replication of templates containing abnormal bases, or possessing structurally aberrant replication forks that inhibit normal DNA synthesis. PMID- 11024017 TI - Munc-18 associates with syntaxin and serves as a negative regulator of exocytosis in the pancreatic beta -cell. AB - The Munc-18 protein (mammalian homologue of the unc-18 gene; also called nSec1 or rbSec1) has been identified as an essential component of the synaptic vesicle fusion protein complex. The cellular and subcellular localization and functional role of Munc-18 protein in pancreatic beta-cells was investigated. Subcellular fractionation of insulin-secreting HIT-T15 cells revealed a 67-kDa protein in both cytosol and membrane fractions. Immunohistochemistry showed punctate Munc-18 immunoreactivity in the cytoplasm of rat pancreatic islet cells. Direct double labeling immunofluorescence histochemistry combined with confocal laser microscopy revealed the presence of Munc-18 immunoreactivity in insulin-, glucagon-, pancreatic polypeptide-, and somatostatin-containing cells. Syntaxin 1 immunoreactivity was detected in extracts of HIT-T15 cells, which were immunoprecipitated using Munc-18 antiserum, suggesting an intimate association of Munc-18 with syntaxin 1. Administration of Munc-18 peptide or Munc-18 antiserum to streptolysin O-permeabilized HIT-T15 cells resulted in significantly increased insulin release, but did not have any significant effect on voltage-gated Ca(2+) channel activity. The findings taken together show that the Munc-18 protein is present in insulin-secreting beta-cells and implicate Munc-18 as a negative regulator of the insulin secretory machinery via a mechanism that does not involve syntaxin-associated Ca(2+) channels. PMID- 11024018 TI - Neutralization of a conserved amino acid residue in the human Na+/glucose transporter (hSGLT1) generates a glucose-gated H+ channel. AB - The role of conserved Asp204 in the human high affinity Na+/glucose cotransporter (hSGLT1) was investigated by site-directed mutagenesis combined with functional assays exploiting the Xenopus oocyte expression system. Substitution of H+ for Na+ reduces the apparent affinity of hSGLT1 for glucose from 0.3 to 6 mm. The apparent affinity for H+ (7 microm) is about three orders of magnitude higher than for Na+ (6 mm). Cation/glucose cotransport exhibits a coupling ratio of 2 Na+ (or 2 H+):1. Pre-steady-state kinetics indicate that similar Na+ - or H+ induced conformational changes are the basis for coupled transport. Replacing Asp204 with Glu increases the apparent affinity for H+ by >20-fold with little impact on the apparent Na+ affinity. This implies that the length of the carboxylate side chain is critical for cation selectivity. Neutralization of Asp204 (Asp --> Asn or Cys) reveals glucose-evoked H(+) currents that were one order of magnitude greater than Na(+) currents. These phlorizin-sensitive H+ currents reverse and are enhanced by internal acidification of oocytes. Together with a H(+) to sugar stoichiometry as high as 145:1, these results favor a glucose-gated H+ channel activity of the mutant. Our observations support the idea that cotransporters and channels share common features. PMID- 11024019 TI - Latrophilin, neurexin, and their signaling-deficient mutants facilitate alpha latrotoxin insertion into membranes but are not involved in pore formation. AB - Pure alpha-latrotoxin is very inefficient at forming channels/pores in artificial lipid bilayers or in the plasma membrane of non-secretory cells. However, the toxin induces pores efficiently in COS-7 cells transfected with the heptahelical receptor latrophilin or the monotopic receptor neurexin. Signaling-deficient (truncated) mutants of latrophilin and latrophilin-neurexin hybrids also facilitate pore induction, which correlates with toxin binding irrespective of receptor structure. This rules out the involvement of signaling in pore formation. With any receptor, the alpha-latrotoxin pores are permeable to Ca(2+) and small molecules including fluorescein isothiocyanate and norepinephrine. Bound alpha-latrotoxin remains on the cell surface without penetrating completely into the cytosol. Higher temperatures facilitate insertion of the toxin into the plasma membrane, where it co-localizes with latrophilin (under all conditions) and with neurexin (in the presence of Ca(2+)). Interestingly, on subsequent removal of Ca(2+), alpha-latrotoxin dissociates from neurexin but remains in the membrane and continues to form pores. These receptor-independent pores are inhibited by anti-alpha-latrotoxin antibodies. Our results indicate that (i) alpha-latrotoxin is a pore-forming toxin, (ii) receptors that bind alpha latrotoxin facilitate its insertion into the membrane, (iii) the receptors are not physically involved in the pore structure, (iv) alpha-latrotoxin pores may be independent of the receptors, and (v) pore formation does not require alpha latrotoxin interaction with other neuronal proteins. PMID- 11024020 TI - Dynamic shuttling of nuclear factor kappa B between the nucleus and cytoplasm as a consequence of inhibitor dissociation. AB - Activation of the nuclear factor kappaB (NFkappaB) transcription factor is intimately associated with its translocation from the cytoplasm to the nucleus. Using the nuclear export inhibitor leptomycin B, we demonstrate shuttling of the RELA subunit of NFkappaB and the inhibitory subunit IkappaBalpha between these two compartments in unstimulated cells. Determination of the kinetics of nuclear entry shows marked differences for the two components; the entry of IkappaBalpha occurs more rapidly than RELA. The shuttling is suggested to be a consequence of the cytoplasmic dissociation of the NFkappaB.IkappaB complex rather than its direct nuclear import or degradation and resynthesis of IkappaBalpha. Using previously published kinetic data, this proposition is born out by the deduction that 17% of NFkappaB is not complexed to IkappaBalpha in a resting cell. A numerical model is presented to validate the proposed regulation of NFkappaB subcellular localization consequent in part on the nuclear export function and in part on the cytoplasmic retention function of IkappaBalpha. We suggest that the non-saturated interaction of NFkappaB with the inhibitor may enhance the specificity of action of IkappaB proteins on different NFkappaB dimers and allow additional modes of regulation of IkappaB function. PMID- 11024021 TI - Identification of two novel RanGTP-binding proteins belonging to the importin beta superfamily. AB - Nucleo-cytoplasmic transport comprises a large number of distinct pathways, many of which are defined by members of the importin beta superfamily of nuclear transport receptors. These transport receptors all directly interact with RanGTP to modulate the compartment-specific binding of their transport substrates. To identify new members of the importin beta family, we used affinity chromatography on immobilized RanGTP and isolated Ran-binding protein (RanBP) 16 from HeLa cell extracts. RanBP16 and its close human homologue, RanBP17, are distant members of the importin beta family. Like the other members of the transport receptor superfamily, RanBP16 interacts with the nuclear pore complex and is able to enter the nucleus independent of energy and additional nuclear transport receptors. PMID- 11024022 TI - AGS3 inhibits GDP dissociation from galpha subunits of the Gi family and rhodopsin-dependent activation of transducin. AB - A number of recently discovered proteins that interact with the alpha subunits of G(i)-like G proteins contain homologous repeated sequences named G protein regulatory (GPR) motifs. Activator of G protein signaling 3 (AGS3), identified as an activator of the yeast pheromone pathway in the absence of the pheromone receptor, has a domain with four such repeats. To elucidate the potential mechanisms of regulation of G protein signaling by proteins containing GPR motifs, we examined the effects of the AGS3 GPR domain on the kinetics of guanine nucleotide exchange and GTP hydrolysis by G(i)alpha(1) and transducin-alpha (G(t)alpha). The AGS3 GPR domain markedly inhibited the rates of spontaneous guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding to G(i)alpha and rhodopsin-stimulated GTPgammaS binding to G(t)alpha. The full-length AGS3 GPR domain, AGS3-(463-650), was approximately 30-fold more potent than AGS3-(572 629), containing two AGS3 GPR motifs. The IC(50) values for the AGS3-(463-650) inhibitory effects on G(i)alpha and transducin were 0.12 and 0.15 microm, respectively. Furthermore, AGS3-(463-650) and AGS3-(572-629) effectively blocked the GDP release from G(i)alpha and rhodopsin-induced dissociation of GDP from G(t)alpha. The potencies of AGS3-(572-629) and AGS3-(463-650) to suppress the GDP dissociation rates correlated with their ability to inhibit the rates of GTPgammaS binding. Consistent with the inhibition of nucleotide exchange, the AGS3 GPR domain slowed the rate of steady-state GTP hydrolysis by G(i)alpha. The catalytic rate of G(t)alpha GTP hydrolysis, measured under single turnover conditions, remained unchanged with the addition of AGS3-(463-650). Altogether, our results suggest that proteins containing GPR motifs, in addition to their potential role as G protein-coupled receptor-independent activators of Gbetagamma signaling pathways, act as GDP dissociation inhibitors and negatively regulate the activation of a G protein by a G protein-coupled receptor. PMID- 11024023 TI - Biochemical characterization of the DNA helicase activity of the escherichia coli RecQ helicase. AB - We demonstrate that RecQ helicase from Escherichia coli is a catalytic helicase whose activity depends on the concentration of ATP, free magnesium ion, and single-stranded DNA-binding (SSB) protein. Helicase activity is cooperative in ATP concentration, with an apparent S(0.5) value for ATP of 200 microm and a Hill coefficient of 3.3 +/- 0.3. Therefore, RecQ helicase utilizes multiple, interacting ATP-binding sites to mediate double-stranded DNA (dsDNA) unwinding, implicating a multimer of at least three subunits as the active unwinding species. Unwinding activity is independent of dsDNA ends, indicating that RecQ helicase can unwind from both internal regions and ends of dsDNA. The K(M) for dsDNA is 0.5-0.9 microm base pairs; the k(cat) for DNA unwinding is 2.3-2.7 base pairs/s/monomer of RecQ helicase; and unexpectedly, helicase activity is optimal at a free magnesium ion concentration of 0.05 mm. Omitting Escherichia coli SSB protein lowers the rate and extent of dsDNA unwinding, suggesting that RecQ helicase associates with the single-stranded DNA (ssDNA) product. In agreement, the ssDNA-dependent ATPase activity is reduced in proportion to the SSB protein concentration; in its absence, ATPase activity saturates at six nucleotides/RecQ helicase monomer and yields a k(cat) of 24 s(-1). Thus, we conclude that SSB protein stimulates RecQ helicase-mediated unwinding by both trapping the separated ssDNA strands after unwinding and preventing the formation of non productive enzyme-ssDNA complexes. PMID- 11024024 TI - Internalization of HIV-1 tat requires cell surface heparan sulfate proteoglycans. AB - Tat, the transactivator protein of human immunodeficiency virus-1, has the unusual capacity of being internalized by cells when present in the extracellular milieu. This property can be exploited for the cellular delivery of heterologous proteins fused to Tat both in cell culture and in living animals. Here we provide genetic and biochemical evidence that cell membrane heparan sulfate (HS) proteoglycans act as receptors for extracellular Tat uptake. Cells genetically defective in the biosynthesis of fully sulfated HS are selectively impaired in the internalization of recombinant Tat fused to the green fluorescent protein, as evaluated by both flow cytometry and functional assays. In wild type cells, Tat uptake is competitively inhibited by soluble heparin and by treatment with glycosaminoglycan lyases specifically degrading HS chains. Cell surface HS proteoglycans also mediate physiological internalization of Tat green fluorescent protein released from neighboring producing cells. In contrast to extracellular Tat uptake, both wild type cells and cells genetically impaired in proteoglycan synthesis are equally proficient in the extracellular release of Tat, thus indicating that proteoglycans are not required for this process. The ubiquitous distribution of HS proteoglycans is consistent with the efficient intracellular delivery of heterologous proteins fused with Tat to different mammalian cell types. PMID- 11024025 TI - Nucleophile selection for the endonuclease activities of human, ovine, and avian retroviral integrases. AB - Retroviral integrases catalyze four endonuclease reactions (processing, joining, disintegration, and nonspecific alcoholysis) that differ in specificity for the attacking nucleophile and target DNA sites. To assess how the two substrates of this enzyme affect each other, we performed quantitative analyses, in three retroviral systems, of the two reactions that use a variety of nucleophiles. The integrase proteins of human immuno- deficiency virus type 1, visna virus, and Rous sarcoma virus exhibited distinct preferences for water or other nucleophiles during site-specific processing of viral DNA and during nonspecific alcoholysis of nonviral DNA. Although exogenous alcohols competed with water as the nucleophile for processing, the alcohols stimulated nicking of nonviral DNA. Moreover, different nucleophiles were preferred when the various integrases acted on different DNA targets. In contrast, the nicking patterns were independent of whether integrase was catalyzing hydrolysis or alcoholysis and were not influenced by the particular exogenous alcohol. Thus, although the target DNA influenced the choice of nucleophile, the nucleophile did not affect the choice of target sites. These results indicate that interaction with target DNA is the critical step before catalysis and suggest that integrase does not reach an active conformation until target DNA has bound to the enzyme. PMID- 11024026 TI - An additional phosphate-binding element in arrestin molecule. Implications for the mechanism of arrestin activation. AB - Arrestins quench the signaling of a wide variety of G protein-coupled receptors by virtue of high-affinity binding to phosphorylated activated receptors. The high selectivity of arrestins for this particular functional form of receptor ensures their timely binding and dissociation. In a continuing effort to elucidate the molecular mechanisms responsible for arrestin's selectivity, we used the visual arrestin model to probe the functions of its N-terminal beta strand I comprising the highly conserved hydrophobic element Val-Ile-Phe (residues 11-13) and the adjacent positively charged Lys(14) and Lys(15). Charge elimination and reversal in positions 14 and 15 dramatically reduce arrestin binding to phosphorylated light-activated rhodopsin (P-Rh*). The same mutations in the context of various constitutively active arrestin mutants (which bind to P Rh*, dark phosphorylated rhodopsin (P-Rh), and unphosphorylated light-activated rhodopsin (Rh*)) have minimum impact on P-Rh* and Rh* binding and virtually eliminate P-Rh binding. These results suggest that the two lysines "guide" receptor-attached phosphates toward the phosphorylation-sensitive trigger Arg(175) and participate in phosphate binding in the active state of arrestin. The elimination of the hydrophobic side chains of residues 11-13 (triple mutation V11A, I12A, and F13A) moderately enhances arrestin binding to P-Rh and Rh*. The effects of triple mutation V11A, I12A, and F13A in the context of phosphorylation independent mutants suggest that residues 11-13 play a dual role. They stabilize arrestin's basal conformation via interaction with hydrophobic elements in arrestin's C-tail and alpha-helix I as well as its active state by interactions with alternative partners. In the context of the recently solved crystal structure of arrestin's basal state, these findings allow us to propose a model of initial phosphate-driven structural rearrangements in arrestin that ultimately result in its transition into the active receptor-binding state. PMID- 11024027 TI - Common requirements for melanocortin-4 receptor selectivity of structurally unrelated melanocortin agonist and endogenous antagonist, Agouti protein. AB - The activity of melanocortin receptors (MCR) is regulated by melanocortin peptide agonists and by the endogenous antagonists, Agouti protein and AgRP (Agouti related protein). To understand how the selectivity for these structurally unrelated agonists and antagonist is achieved, chimeric and mutants MC3R and MC4R were expressed in cell lines and pharmacologically analyzed. A region containing the third extracellular loop, EC3, of MC4R was essential for selective Agouti protein antagonism. In addition, this part of MC4R, when introduced in MC3R, conferred Agouti protein antagonism. Further mutational analysis of this region of MC4R demonstrated that Tyr(268) was required for the selective interaction with Agouti protein, because a profound loss of the ability of Agouti protein to inhibit (125)I-labeled [Nle(4),d-Phe(7)]alpha-melanocyte-stimulating hormone (MSH) binding was observed by the single mutation of Tyr(268) to Ile. This same residue conferred selectivity for the MC4R selective agonist, [d-Tyr(4)]MT-II, whereas it inhibited interaction with the MC3R-selective agonist, [Nle(4)]Lys gamma(2)-MSH. Conversely, mutation of Ile(265) in MC3 (the corresponding residue of Tyr(268)) to Tyr displayed a gain of affinity for [d-Tyr(4)]MT-II, but not for Agouti protein, and a loss of affinity for [Nle(4)]Lys-gamma(2)-MSH as compared with wild-type MC3R. This single amino acid mutation thus confers the selectivity of MC3R toward a pharmacological profile like that observed for MC4R agonists but not for the antagonist, Agouti protein. Thus, selectivity for structurally unrelated ligands with opposite activities is achieved in a similar manner for MC4R but not for MC3R. PMID- 11024028 TI - Rh type B glycoprotein is a new member of the Rh superfamily and a putative ammonia transporter in mammals. AB - Ammonium transporters play a key functional role in nitrogen uptake and assimilation in microorganisms and plants; however, little is known about their structural counterpart in mammals. Here, we report the molecular cloning and biochemical characterization of Rh type B glycoproteins, human RhBG and mouse Rhbg, two new members of the Rh family with distinct tissue specificities. The RhBG orthologues possess a conserved 12-transmembrane topology and most resemble bacterial and archaeal ammonium transporters. Human RHBG resides at chromosome 1q21.3, which harbors candidate genes for medullary cystic kidney disease, whereas mouse Rhbg is syntenic on chromosome 3. Northern blot and in situ hybridization revealed that RHBG and Rhbg are predominantly expressed in liver, kidney, and skin, the specialized organs involving ammonia genesis, excretion, or secretion. Confocal microscopy showed that RhBG is located in the plasma membrane and in some intracellular granules. Western blots of membrane proteins from stable HEK293 cells and from mouse kidney and liver confirmed this distribution. N-Glycanase digestion showed that RhBG/Rhbg has a carbohydrate moiety probably attached at the NHS motif on exoloop 1. Phylogenetic clustering, tissue-specific expression, and plasma membrane location suggest that RhBG homologous proteins are the long sought major ammonium transporters in mammalians. PMID- 11024029 TI - Mice with a deletion in the gene for CCAAT/enhancer-binding protein beta have an attenuated response to cAMP and impaired carbohydrate metabolism. AB - Fifty percent of the mice homozygous for a deletion in the gene for CCAAT/enhancer-binding protein beta (C/EBP beta-/- mice; B phenotype) die within 1 to 2 h after birth of hypoglycemia. They do not mobilize their hepatic glycogen or induce the cytosolic form of phosphoenolpyruvate carboxykinase (PEPCK). Administration of cAMP resulted in mobilization of glycogen, induction of PEPCK mRNA, and a normal blood glucose; these mice survived beyond 2 h postpartum. Adult C/EBP beta-/- mice (A phenotype) also had difficulty in maintaining blood glucose levels during starvation. Fasting these mice for 16 or 30 h resulted in lower levels of hepatic PEPCK mRNA, blood glucose, beta-hydroxybutyrate, blood urea nitrogen, and gluconeogenesis when compared with control mice. The concentration of hepatic cAMP in these mice was 50% of controls, but injection of theophylline, together with glucagon, resulted in a normal cAMP levels. Agonists (glucagon, epinephrine, and isoproterenol) and other effectors of activation of adenylyl cyclase were the same in liver membranes isolated from C/EBP beta-/- mice and littermates. The hepatic activity of cAMP-dependent protein kinase was 80% of wild type mice. There was a 79% increase in the concentration of RI alpha and 27% increase in RII alpha in the particulate fraction of the livers of C/EBP beta-/- mice relative to wild type mice, with no change in the catalytic subunit (C alpha). Thus, a 45% increase in hepatic cAMP (relative to the wild type) would be required in C/EBP beta-/- mice to activate protein kinase A by 50%. In addition, the total activity of phosphodiesterase in the livers of C/EBP beta-/- mice, as well as the concentration of mRNA for phosphodiesterase 3A (PDE3A) and PDE3B was approximately 25% higher than in control animals, suggesting accelerated degradation of cAMP. C/EBP beta influences the regulation of carbohydrate metabolism by altering the level of hepatic cAMP and the activity of protein kinase A. PMID- 11024030 TI - Binding of an RNA trafficking response element to heterogeneous nuclear ribonucleoproteins A1 and A2. AB - Heterogeneous nuclear ribonucleoprotein (hnRNP) A2 binds a 21-nucleotide myelin basic protein mRNA response element, the A2RE, and A2RE-like sequences in other localized mRNAs, and is a trans-acting factor in oligodendrocyte cytoplasmic RNA trafficking. Recombinant human hnRNPs A1 and A2 were used in a biosensor to explore interactions with A2RE and the cognate oligodeoxyribonucleotide. Both proteins have a single site that bound oligonucleotides with markedly different sequences but did not bind in the presence of heparin. Both also possess a second, specific site that bound only A2RE and was unaffected by heparin. hnRNP A2 bound A2RE in the latter site with a K(d) near 50 nm, whereas the K(d) for hnRNP A1 was above 10 microm. UV cross-linking assays led to a similar conclusion. Mutant A2RE sequences, that in earlier qualitative studies appeared not to bind hnRNP A2 or support RNA trafficking in oligodendrocytes, had dissociation constants above 5 microm for this protein. The two concatenated RNA recognition motifs (RRMs), but not the individual RRMs, mimicked the binding behavior of hnRNP A2. These data highlight the specificity of the interaction of A2RE with these hnRNPs and suggest that the sequence-specific A2RE-binding site on hnRNP A2 is formed by both RRMs acting in cis. PMID- 11024031 TI - The function of Arg-94 in the oxidation and decarboxylation of glutaryl-CoA by human glutaryl-CoA dehydrogenase. AB - Glutaryl-CoA dehydrogenase catalyzes the oxidation and decarboxylation of glutaryl-CoA to crotonyl-CoA and CO(2). Inherited defects in the protein cause glutaric acidemia type I, a fatal neurologic disease. Glutaryl-CoA dehydrogenase is the only member of the acyl-CoA dehydrogenase family with a cationic residue, Arg-94, situated in the binding site of the acyl moiety of the substrate. Crystallographic investigations suggest that Arg-94 is within hydrogen bonding distance of the gamma-carboxylate of glutaryl-CoA. Substitution of Arg-94 by glycine, a disease-causing mutation, and by glutamine, which is sterically more closely related to arginine, reduced k(cat) of the mutant dehydrogenases to 2-3% of k(cat) of the wild type enzyme. K(m) of these mutant dehydrogenases for glutaryl-CoA increases 10- to 16-fold. The steady-state kinetic constants of alternative substrates, hexanoyl-CoA and glutaramyl-CoA, which are not decarboxylated, are modestly affected by the mutations. The latter changes are probably due to steric and polar effects. The dissociation constants of the non oxidizable substrate analogs, 3-thiaglutaryl-CoA and acetoacetyl-CoA, are not altered by the mutations. However, abstraction of a alpha-proton from 3 thiaglutaryl-CoA, to yield a charge transfer complex with the oxidized flavin, is severely limited. In contrast, abstraction of the alpha-proton of acetoacetyl-CoA by Arg-94 --> Gln mutant dehydrogenase is unaffected, and the resulting enolate forms a charge transfer complex with the oxidized flavin. These experiments indicate that Arg-94 does not make a major contribution to glutaryl-CoA binding. However, the electric field of Arg-94 may stabilize the dianions resulting from abstraction of the alpha-proton of glutaryl-CoA and 3-thiaglutaryl-CoA, both of which contain gamma-carboxylates. It is also possible that Arg-94 may orient glutaryl-CoA and 3-thiaglutaryl-CoA for abstraction of an alpha-proton. PMID- 11024032 TI - Characterization of Fyn-mediated tyrosine phosphorylation sites on GluR epsilon 2 (NR2B) subunit of the N-methyl-D-aspartate receptor. AB - The N-methyl-d-aspartate (NMDA) receptors play critical roles in synaptic plasticity, neuronal development, and excitotoxicity. Tyrosine phosphorylation of NMDA receptors by Src-family tyrosine kinases such as Fyn is implicated in synaptic plasticity. To precisely address the roles of NMDA receptor tyrosine phosphorylation, we identified Fyn-mediated phosphorylation sites on the GluR epsilon 2 (NR2B) subunit of NMDA receptors. Seven out of 25 tyrosine residues in the C-terminal cytoplasmic region of GluR epsilon 2 were phosphorylated by Fyn in vitro. Of these 7 residues, Tyr-1252, Tyr-1336, and Tyr-1472 in GluR epsilon 2 were phosphorylated in human embryonic kidney fibroblasts when co-expressed with active Fyn, and Tyr-1472 was the major phosphorylation site in this system. We then generated rabbit polyclonal antibodies specific to Tyr-1472-phosphorylated GluR epsilon 2 and showed that Tyr-1472 of GluR epsilon 2 was indeed phosphorylated in murine brain using the antibodies. Importantly, Tyr-1472 phosphorylation was greatly reduced in fyn mutant mice. Moreover, Tyr-1472 phosphorylation became evident when hippocampal long term potentiation started to be observed, and its magnitude became larger in murine brain. Finally, Tyr-1472 phosphorylation was significantly enhanced after induction of long term potentiation in the hippocampal CA1 region. These data suggest that Tyr-1472 phosphorylation of GluR epsilon 2 is important for synaptic plasticity. PMID- 11024033 TI - Identification of the gamma subunit-interacting residues on photoreceptor cGMP phosphodiesterase, PDE6alpha '. AB - Photoreceptor cGMP phosphodiesterase (PDE6) is the effector enzyme in the G protein-mediated visual transduction cascade. In the dark, the activity of PDE6 is shut off by the inhibitory gamma subunit (Pgamma). Chimeric proteins between cone PDE6alpha' and cGMP-binding and cGMP-specific PDE (PDE5) have been constructed and expressed in Sf9 cells to study the mechanism of inhibition of PDE6 catalytic activity by Pgamma. Substitution of the segment PDE5-(773-820) by the corresponding PDE6alpha'-(737-784) sequence in the wild-type PDE5 or in a PDE5/PDE6alpha' chimera containing the catalytic domain of PDE5 results in chimeric enzymes capable of inhibitory interaction with Pgamma. The catalytic properties of the chimeric PDEs remained similar to those of PDE5. Ala-scanning mutational analysis of the Pgamma-binding region, PDE6alpha'-(750-760), revealed PDE6alpha' residues essential for the interaction. The M758A mutation markedly impaired and the Q752A mutation moderately impaired the inhibition of chimeric PDE by Pgamma. The analysis of the catalytic properties of mutant PDEs and a model of the PDE6 catalytic domain suggest that residues Met(758) and Gln(752) directly bind Pgamma. A model of the PDE6 catalytic site shows that PDE6alpha' (750-760) forms a loop at the entrance to the cGMP-binding pocket. Binding of Pgamma to Met(758) would effectively block access of cGMP to the catalytic cavity, providing a structural basis for the mechanism of PDE6 inhibition. PMID- 11024034 TI - Expression of the transcription factor STAT-1 alpha in insulinoma cells protects against cytotoxic effects of multiple cytokines. AB - Destruction of pancreatic islet beta-cells in type 1 diabetes appears to result from direct contact with infiltrating T-cells and macrophages and exposure to inflammatory cytokines such as interferon (IFN)-gamma, interleukin (IL)-1 beta, and tumor necrosis factor TNF-alpha that such cells produce. We recently reported on a method for selection of insulinoma cells that are resistant to the cytotoxic effects of inflammatory cytokines (INS-1(res)), involving their growth in progressively increasing concentrations of IL-1 beta plus IFN-gamma, and selection of surviving cells. In the current study, we have investigated the molecular mechanism of cytokine resistance in INS-1(res) cells. By focusing on the known components of the IFN-gamma receptor signaling pathway, we have discovered that expression levels of signal transducer and activator of transcription (STAT)-1 alpha are closely correlated with the cytokine-resistant and -sensitive phenotypes. That STAT-1 alpha is directly involved in development of cytokine resistance is demonstrated by an increase of viability from 10 +/- 2% in control cells to 50 +/- 6% in cells with adenovirus-mediated overexpression of STAT-1 alpha (p < 0.001) after culture of both cell groups in the presence of 100 units/ml IFN-gamma plus 10 ng/ml IL-1 beta for 48 h. The resistance to IL-1 beta plus IFN-gamma in STAT-1 alpha-expressing cells is due in part to interference with IL-1 beta-mediated stimulation of inducible nitric-oxide synthase expression and nitric oxide production. Furthermore, overexpression of STAT-1 alpha does not impair robust glucose-stimulated insulin secretion in the INS-1-derived cell line 832/13. We conclude that expression of STAT-1 alpha may be a means of protecting insulin-producing cell lines from cytokine damage, which, in conjunction with appropriate cell-impermeant macroencapsulation devices, may allow such cells to be used for insulin replacement in type 1 diabetes. PMID- 11024035 TI - Transcription factors recognizing overlapping C1-A2 binding sites positively regulate insulin gene expression. AB - Transcription factors binding the insulin enhancer region, RIPE3b, mediate beta cell type-specific and glucose-responsive expression of the insulin gene. Earlier studies demonstrate that activator present in the beta-cell-specific RIPE3b1 binding complex is critical for these actions. The DNA binding activity of the RIPE3b1 activator is induced in response to glucose stimulation and is inhibited under glucotoxic conditions. The C1 element within the RIPE3b region has been implicated as the binding site for RIPE3b1 activator. The RIPE3b region also contains an additional element, A2, which shares homology with the A elements in the insulin enhancer. Transcription factors (PDX-1 and HNF-1 alpha) binding to A elements are critical regulators of insulin gene expression and/or pancreatic development. Hence, to understand the roles of C1 and A2 elements in regulating insulin gene expression, we have systematically mutated the RIPE3b region and analyzed the effect of these mutations on gene expression. Our results demonstrate that both C1 and A2 elements together constitute the binding site for the RIPE3b1 activator. In addition to C1-A2 (RIPE3b) binding complexes, three binding complexes that specifically recognize A2 elements are found in nuclear extracts from insulinoma cell lines; the A2.2 complex is detected only in insulin producing cell lines. Furthermore, two base pairs in the A2 element were critical for binding of both RIPE3b1 and A2.2 activators. Transient transfection results indicate that both C1-A2 and A2-specific binding activators cooperatively activate insulin gene expression. In addition, RIPE3b1- and A2-specific activators respond differently to glucose, suggesting that their overlapping binding specificity and functional cooperation may play an important role in regulating insulin gene expression. PMID- 11024036 TI - CCAAT/enhancer-binding protein alpha is required for transcription of the beta 3 adrenergic receptor gene during adipogenesis. AB - The beta(3)-adrenergic receptor (beta(3)AR) is expressed predominantly in adipocytes, and it plays a major role in regulating lipolysis and adaptive thermogenesis. Its expression in a variety of adipocyte cell models is preceded by the appearance of CCAAT/enhancer-binding protein alpha (C/EBP alpha), which has been shown to regulate a number of other adipocyte-specific genes. Importantly, it has been demonstrated that several adipocyte cell lines that fail to express C/EBP alpha exhibit reduced insulin sensitivity, despite an apparent adipogenic phenotype. Here we show that transcription and function of the beta(3)AR correlates with C/EBP alpha expression in these adipocyte models. A 5.13-kilobase pair fragment of the mouse beta(3)AR promoter was isolated and sequenced. This fragment conferred a 50-fold increase in luciferase reporter gene expression in adipocytes. Two putative C/EBP binding sites exist at -3306 to 3298 and at -1462 to -1454, but only the more distal site is functional. Oligonucleotides corresponding to both the wild-type and mutated -3306 element were inserted upstream of a thymidine kinase luciferase construct. When cotransfected in fibroblasts with a C/EBP alpha expression vector, reporter gene expression increased 3-fold only in the wild-type constructs. The same mutation, when placed into the intact 5.13-kilobase pair promoter, reduced promoter activity in adipocytes from 50-fold to <10-fold. Electrophoretic mobility shift analysis demonstrated that the site at -3306 generated a specific protein oligonucleotide complex that was supershifted by C/EBP alpha antibody, while a probe corresponding to a putative site at -1462 did not. These results define C/EBP alpha as a key transcriptional regulator of the mouse beta(3)AR gene during adipogenesis. PMID- 11024037 TI - Regulation of Cbl molecular interactions by the co-receptor molecule CD43 in human T cells. AB - CD43, one of the most abundant glycoproteins on the T cell surface, has been implicated in selection and maturation of thymocytes and migration, adhesion, and activation of mature T cells. The adapter molecule Cbl has been shown to be a negative regulator of Ras. Furthermore, it may also regulate intracellular signaling through the formation of several multi-molecular complexes. Here we investigated the role of Cbl in the CD43-mediated signaling pathway in human T cells. Unlike T cell receptor signaling, the interaction of the adapter protein Cbl with Vav and phosphatidylinositol 3-kinase, resulting from CD43-specific signals, is independent of Cbl tyrosine phosphorylation, suggesting an alternative mechanism of interaction. CD43 signals induced a Cbl serine phosphorylation-dependent interaction with the tau-isoform of 14-3-3. protein. Protein kinase C-mediated Cbl serine phosphorylation was required for this interaction, because the PKC inhibitor RO-31-8220 prevented it, as well as 14-3-3 dimerization. Moreover, mutation of Cbl serine residues 619, 623, 639, and 642 abolished the interaction between Cbl and 14-3-3. Overexpression of Cbl in Jurkat cells inhibited the CD43-dependent activation of the mitogen-activated protein kinase (MAPK) pathway and AP-1 transcriptional activity, confirming nevertheless a negative role for Cbl in T cell signaling. However, under normal conditions, PKC activation resulting from CD43 engagement was required to activate the MAPK pathway, suggesting that phosphorylation of Cbl on serine residues by PKC and its association with 14-3-3 molecules may play a role in preventing the Cbl inhibitory effect on the Ras-MAPK pathway. These data suggest that by inducing its phosphorylation on serine residues, CD43-mediated signals may regulate the molecular associations and functions of the Cbl adapter protein. PMID- 11024038 TI - Acquisition of Lubrol insolubility, a common step for growth hormone and prolactin in the secretory pathway of neuroendocrine cells. AB - Rat prolactin in the dense cores of secretory granules of the pituitary gland is a Lubrol-insoluble aggregate. In GH(4)C(1) cells, newly synthesized rat prolactin and growth hormone were soluble, but after 30 min about 40% converted to a Lubrol insoluble form. Transport from the endoplasmic reticulum is necessary for conversion to Lubrol insolubility, since incubating cells with brefeldin A or at 15 degrees C reduced formation of insoluble rat (35)S-prolactin. Formation of Lubrol-insoluble aggregates has protein and cell specificity; newly synthesized human growth hormone expressed in AtT20 cells underwent a 40% conversion to Lubrol insolubility with time, but albumin did not, and human growth hormone expressed in COS cells underwent less than 10% conversion to Lubrol insolubility. del32-46 growth hormone, a naturally occurring form of growth hormone, and P89L growth hormone underwent conversion, although they were secreted more slowly, indicating that there is some tolerance in structural requirements for aggregation. An intracellular compartment with an acidic pH is not necessary for conversion to Lubrol insolubility, because incubation with chloroquine or bafilomycin slowed, but did not prevent, the conversion. GH(4)C(1) cells treated with estradiol, insulin, and epidermal growth factor accumulate more secretory granules and store more prolactin, but not more growth hormone, than untreated cells; Lubrol-insoluble aggregates of prolactin and growth hormone formed to the same extent in hormone-treated or untreated GH(4)C(1) cells, but prolactin was retained longer in hormone-treated cells. These findings indicate that aggregation alone is not sufficient to cause retention of secretory granule proteins, and there is an additional selective process. PMID- 11024039 TI - The high light-inducible polypeptides in Synechocystis PCC6803. Expression and function in high light. AB - There are five Synechocystis PCC6803 genes encoding polypeptides with similarity to the Lhc polypeptides of plants. Four of the polypeptides, designated HliA-D (Dolganov, N. A. M., Bhaya, D., and Grossman, A. R. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 636-640) (corresponding to ScpC, ScpD, ScpB, and ScpE in Funk, C., and Vermaas, W. (1999) Biochemistry 38, 9397-9404) contain a single transmembrane domain. The fifth polypeptide (HemH) represents a fusion between a ferrochelatase and an Hli-like polypeptide. By using an epitope tag to identify specifically the different Hli polypeptides, the accumulation of each (excluding HemH) was examined under various environmental conditions. The levels of all of the Hli polypeptides were elevated in high light and during nitrogen limitation, whereas HliA, HliB, and HliC also accumulated to high levels following exposure to sulfur deprivation and low temperature. The temporal pattern of accumulation was significantly different among the different Hli polypeptides. HliC rapidly accumulated in high light, and its level remained high for at least 24 h. HliA and HliB also accumulated rapidly, but their levels began to decline 9-12 h following the imposition of high light. HliD was transiently expressed in high light and was not detected 24 h after the initiation of high light exposure. These results demonstrate that there is specificity to the accumulation of the Hli polypeptides under a diverse range of environmental conditions. Furthermore, mutants for the individual and combinations of the hli genes were evaluated for their fitness to grow in high light. Although all of the mutants grew as fast as wild-type cells in low light, strains inactivated for hliA or hliC/hliD were unable to compete with wild-type cells during co-cultivation in high light. A mutant lacking all four hli genes gradually lost its photosynthesis capacity and died in high light. Hence, the Hli polypeptides are critical for survival when Synechocystis PCC6803 is absorbing excess excitation energy and may allow the cells to cope more effectively with the production of reactive oxygen species. PMID- 11024040 TI - The transcriptional activator Cat8p provides a major contribution to the reprogramming of carbon metabolism during the diauxic shift in Saccharomyces cerevisiae. AB - In yeast, the transition between the fermentative and the oxidative metabolism, called the diauxic shift, is associated with major changes in gene expression and protein synthesis. The zinc cluster protein Cat8p is required for the derepression of nine genes under nonfermentative growth conditions (ACS1, FBP1, ICL1, IDP2, JEN1, MLS1, PCK1, SFC1, and SIP4). To investigate whether the transcriptional control mediated by Cat8p can be extended to other genes and whether this control is the main control for the changes in the synthesis of the respective proteins during the adaptation to growth on ethanol, we analyzed the transcriptome and the proteome of a cat8 Delta strain during the diauxic shift. In this report, we demonstrate that, in addition to the nine genes known as Cat8p dependent, there are 25 other genes or open reading frames whose expression at the diauxic shift is altered in the absence of Cat8p. For all of the genes characterized here, the Cat8p-dependent control results in a parallel alteration in mRNA and protein synthesis. It appears that the biochemical functions of the proteins encoded by Cat8p-dependent genes are essentially related to the first steps of ethanol utilization, the glyoxylate cycle, and gluconeogenesis. Interestingly, no function involved in the tricarboxylic cycle and the oxidative phosphorylation seems to be controlled by Cat8p. PMID- 11024041 TI - Low pH-induced conformational changes in vesicular stomatitis virus glycoprotein involve dramatic structure reorganization. AB - Membrane fusion is the key step in the entry of enveloped animal viruses into their host cells. Fusion of vesicular stomatitis virus with membranes occurs at acidic pH and is mediated by its envelope glycoprotein, the G protein. To study the structural transitions induced by acidic pH on G protein, we have extracted the protein from purified virus by incubation with nonionic detergent. At pH 6.0, purified G protein was able to mediate fusion of either phospholipid vesicles or Vero cells in culture. Intrinsic fluorescence studies revealed that changes in the environment of Trp residues occurred as pH decreases. In the absence of lipidic membranes, acidification led to G protein aggregation, whereas protein protein interactions were substituted by protein-lipid interactions in the presence of liposomes. 1,1'-Bis(4-aniline-5-naphthalene sulfonate) (bis-ANS) binding was utilized to probe the degree of exposure of hydrophobic regions of G protein during acidification. Bis-ANS binding was maximal at pH 6.2, suggesting that a hydrophobic segment is exposed to the medium at this pH. At pH 6.0, a dramatic decrease in bis-ANS binding was observed, probably due to loss of tridimensional structure during the conformational rearrangement. This hypothesis was confirmed by circular dichroism analysis at different pH values, which showed a great decrease in alpha-helix content at pH values close to 6.0, suggesting that a reorganization of G protein secondary structure occurs during the fusion reaction. Our results indicate that G protein undergoes dramatic structural changes at acidic pH and acquires a conformational state able to interact with the target membrane. PMID- 11024042 TI - The translational regulation of lipoprotein lipase in diabetic rats involves the 3'-untranslated region of the lipoprotein lipase mRNA. AB - Adipose tissue lipoprotein lipase (LPL) activity is decreased in patients with poorly controlled diabetes, and this contributes to the dyslipidemia of diabetes. To study the mechanism of this decrease in LPL, we studied adipose tissue LPL expression in male rats with streptozotocin-induced diabetes. Heparin releasable and extractable LPL activity in the epididymal fat decreased by 75-80% in the diabetic group and treatment of the rats with insulin prior to sacrifice reversed this effect. Northern blot analysis indicated no corresponding change in LPL mRNA levels. However, LPL synthetic rate, measured using [(35)S]methionine pulse labeling, was decreased by 75% in the diabetic adipocytes, and insulin treatment reversed this effect. These results suggested regulation of LPL at the level of translation. Diabetic adipocytes demonstrated no change in the distribution of LPL mRNA associated with polysomes, suggesting no inhibition of translation initiation. Addition of cytoplasmic extracts from control and diabetic adipocytes to a reticulocyte lysate system demonstrated the inhibition of LPL translation in vitro. Using different LPL mRNA transcripts in this in vitro translation assay, we found that the 3'-untranslated region (UTR) of the LPL mRNA was important in controlling translation inhibition by the cytoplasmic extracts. To identify the specific region involved, gel shift analysis was performed. A specific shift in mobility was observed when diabetic cytoplasmic extract was added to a transcript containing nucleotides 1818-2000 of the LPL 3'-UTR. Thus, inhibition of translation is the predominant mechanism for the decreased adipose tissue LPL in this insulin-deficient model of diabetes. Translation inhibition involves the interaction of a cytoplasmic factor, probably an RNA-binding protein, with specific sequences of the LPL 3'-UTR. PMID- 11024043 TI - DNA structure and aspartate 276 influence nucleotide binding to human DNA polymerase beta. Implication for the identity of the rate-limiting conformational change. AB - Structures of DNA polymerase (pol) beta bound to single-nucleotide gapped DNA had revealed that the lyase and pol domains form a "doughnut-shaped" structure altering the dNTP binding pocket in a fashion that is not observed when bound to non-gapped DNA. We have investigated dNTP binding to pol beta-DNA complexes employing steady-state and pre-steady-state kinetics. Although pol beta has a kinetic scheme similar to other DNA polymerases, polymerization by pol beta is limited by at least two partially rate-limiting steps: a conformational change after dNTP ground-state binding and product release. The equilibrium binding constant, K(d)((dNTP)), decreased and the insertion efficiency increased with a one-nucleotide gapped DNA substrate, as compared with non-gapped DNA. Valine substitution for Asp(276), which interacts with the base of the incoming nucleotide, increased the binding affinity for the incoming nucleotide indicating that the negative charge contributed by Asp(276) weakens binding and that an interaction between residue 276 with the incoming nucleotide occurs during ground state binding. Since the interaction between Asp(276) and the nascent base pair is observed only in the "closed" conformation of pol beta, the increased free energy in ground-state binding for the mutant suggests that the subsequent rate limiting conformational change is not the "open" to "closed" structural transition, but instead is triggered in the closed pol conformation. PMID- 11024044 TI - Reduction of isoprostanes and regression of advanced atherosclerosis by apolipoprotein E. AB - Apolipoprotein E is a multifunctional protein synthesized by hepatocytes and macrophages. Plasma apoE is largely liver-derived and known to regulate lipoprotein metabolism. Macrophage-derived apoE has been shown to reduce the progression of atherosclerosis in mice. We tested the hypothesis that liver derived apoE could directly induce regression of pre-existing advanced atherosclerotic lesions without reducing plasma cholesterol levels. Aged low density lipoprotein (LDL) receptor-deficient (LDLR(-/-)) mice were fed a western type diet for 14 weeks to induce advanced atherosclerotic lesions. One group of mice was sacrificed for evaluation of atherosclerosis at base line, and two other groups were injected with a second generation adenoviruses encoding human apoE3 or a control empty virus. Hepatic apoE gene transfer increased plasma apoE levels by 4-fold at 1 week, and apoE levels remained at least 2-fold higher than controls at 6 weeks. There were no significant changes in plasma total cholesterol levels or lipoprotein composition induced by expression of apoE. The liver-derived human apoE gained access to and was retained in arterial wall. Compared with base-line mice, the control group demonstrated progression of atherosclerosis; in contrast, hepatic apoE expression induced highly significant regression of advanced atherosclerotic lesions. Regression of lesions was accompanied by the loss of macrophage-derived foam cells and a trend toward increase in extracellular matrix of lesions. As an index of in vivo oxidant stress, we quantitated the isoprostane iPF(2 alpha)-VI and found that expression of apoE markedly reduced urinary, LDL-associated, and arterial wall iPF(2 alpha) VI levels. In summary, these results demonstrate that liver-derived apoE directly induced regression of advanced atherosclerosis and has anti-oxidant properties in vivo that may contribute to its anti-atherogenic effects. PMID- 11024045 TI - Livin, a novel inhibitor of apoptosis protein family member. AB - A novel human inhibitor of apoptosis protein (IAP) family member termed Livin was identified, containing a single baculoviral IAP repeat (BIR) domain and a COOH terminal RING finger domain. The mRNA for livin was not detectable by Northern blot in most normal adult tissues with the exception of the placenta, but was present in developmental tissues and in several cancer cell lines. Highest levels were observed in two melanoma-derived cell lines, G361 and SK-Mel29. Transfection of livin in HeLa cells resulted in protection from apoptosis induced by expression of FADD, Bax, RIP, RIP3, and DR6. Similar to other IAP family members, the anti-apoptotic activity of Livin was dependent on the BIR domain. Livin was also capable of inhibiting DEVD-like caspase activity triggered by tumor necrosis factor-alpha. In vitro binding studies demonstrated a direct interaction between Livin and the active form of the downstream caspases, caspase-3 and -7, that was dependent on the BIR domain of Livin. In addition, the unprocessed and cleaved forms of caspase-9 co-immunoprecipitated with Livin in vivo, and recombinant Livin could inhibit the activation of caspase-9 induced by Apaf-1, cytochrome c, and dATP. The subcellular distribution of the transfected Livin was analyzed by immunofluorescence. Both Livin and Survivin were expressed in the nucleus and in a filamentous pattern throughout the cytoplasm. In contrast to the apoptotic activity, the COOH-terminal RING domain mediated its subcellular localization patterning. Further studies found that transfection of an antisense construct against livin could trigger apoptosis specifically in cell lines expressing livin mRNA. This was associated with an increase in DNA fragmentation and in DEVD-like caspase activity. Thus, disruption of Livin may provide a strategy to induce apoptosis in certain cancer cells. PMID- 11024046 TI - Platelet glycoprotein Ib alpha binds to thrombin anion-binding exosite II inducing allosteric changes in the activity of thrombin. AB - The glycoprotein (GP) Ib-IX complex is a platelet surface receptor that binds thrombin as one of its ligands, although the biological significance of thrombin interaction remains unclear. In this study we have used several approaches to investigate the GPIb alpha-thrombin interaction in more detail and to study its effect on the thrombin-induced elaboration of fibrin. We found that both glycocalicin and the amino-terminal fragment of GPIb alpha reduced the release of fibrinopeptide A from fibrinogen by about 50% by a noncompetitive allosteric mechanism. Similarly, GPIb alpha caused in thrombin an allosteric reduction in the rate of turnover of the small peptide substrate d-Phe-Pro-Arg-pNA. The K(d) for the glycocalicin-thrombin interaction was 1 microm at physiological ionic strength but was highly salt-dependent, decreasing to 0.19 microm at 100 mm NaCl (Gamma(salt) = -4.2). The salt dependence was characteristic of other thrombin ligands that bind to exosite II of this enzyme, and we confirmed this as the GPIb alpha-binding site on thrombin by using thrombin mutants and by competition binding studies. R68E or R70E mutations in exosite I of thrombin had little effect on its interaction with GPIb alpha. Both the allosteric inhibition of fibrinogen turnover caused by GPIb alpha binding to these mutants, and the K(d) values for their interactions with GPIb alpha were similar to those of wild-type thrombin. In contrast, R89E and K248E mutations in exosite II of thrombin markedly increased the K(d) values for the interactions of these thrombin mutants with GPIb alpha by 10- and 25-fold, respectively. Finally, we demonstrated that low molecular weight heparin (which binds to thrombin exosite II) but not hirugen (residues 54-65 of hirudin, which binds to exosite I of thrombin) inhibited thrombin binding to GPIb alpha. These data demonstrate that GPIb alpha binds to thrombin exosite II and in so doing causes a conformational change in the active site of thrombin by an allosteric mechanism that alters the accessibility of both its natural substrate, fibrinogen, and the small peptidyl substrate d-Phe-Pro-Arg pNA. PMID- 11024047 TI - Nodal signaling uses activin and transforming growth factor-beta receptor regulated Smads. AB - Nodal, a member of the transforming growth factor beta (TGF-beta) superfamily, is implicated in many events critical to the early vertebrate embryo, including mesoderm formation, anterior patterning, and left-right axis specification. Here we define the intracellular signaling pathway induced by recombinant nodal protein treatment of P19 embryonal carcinoma cells. Nodal signaling activates pAR3-Lux, a luciferase reporter previously shown to respond specifically to activin and TGF-beta. However, nodal is unable to induce pTlx2-Lux, a reporter specifically responsive to bone morphogenetic proteins. We also demonstrate that nodal induces p(CAGA)(12), a reporter previously shown to be specifically activated by Smad3. Expression of a dominant negative Smad2 significantly reduces the level of luciferase reporter activity induced by nodal treatment. Finally, we show that nodal signaling rapidly leads to the phosphorylation of Smad2. These results provide the first direct biochemical evidence that nodal signaling is mediated by both activin-TGF-beta pathway Smads, Smad2 and Smad3. We also show here that the extracellular cripto protein is required for nodal signaling, making it distinct from activin or TGF-beta signaling. PMID- 11024048 TI - The critical role of the stem region as a functional domain responsible for the oligomerization and Golgi localization of N-acetylglucosaminyltransferase V. The involvement of a domain homophilic interaction. AB - We demonstrated that a region in the stem of N-acetylglucosaminyltransferase V (GnT-V), a Golgi resident protein, is not required for enzyme activity but serves as functional domain, responsible for intracellular localization. Deletion of the domain led to complete retention of the kinetic properties but resulted in the cell surface localization of the enzyme as well as its efficient secretion into the medium. The lack of this domain concomitantly abolished the disulfide mediated oligomerization of GnT-V, which appears to confer the Golgi retention. When the domain was inserted into the stem region of a cell surface-localized type II membrane protein, the resulting chimeric protein was substantially oligomerized and predominantly localized in the intracellular organelle. Furthermore, it was found that the presence of this domain is exclusively responsible for homo-oligomer formation. This homophilic interaction appears to involve a hydrophobic cluster of residues in the alpha-helix of the domain, as indicated by secondary structure predictions. These findings suggest that the domain specifically participates in the Golgi retention of GnT-V, probably via inducing homo-oligomer formation, and would also provide a possible mechanism for the oligomerization, which is critical for localization in the Golgi. PMID- 11024049 TI - Regulation of RNA polymerase III transcription during cell cycle entry. AB - Increased rates of RNA polymerase (pol) III transcription constitute a central feature of the mitogenic response, but little is known about the mechanism(s) responsible. We demonstrate that the retinoblastoma protein RB plays a major role in suppressing pol III transcription in growth-arrested fibroblasts. RB knockout cells are compromised in their ability to down-regulate pol III following serum withdrawal. RB binds and represses the pol III-specific transcription factor TFIIIB during G(0) and early G(1), but this interaction decreases as cells approach S phase. Full induction of pol III coincides with mid- to late G(1) phase, when RB becomes phosphorylated by cyclin D- and E-dependent kinases. TFIIIB only associates with the underphosphorylated form of RB, and overexpression of cyclins D and E stimulates pol III transcription in vivo. The RB-related protein p130 also contributes to the repression of TFIIIB in growth arrested fibroblasts. These observations provide insight into the mechanisms responsible for controlling pol III transcription during the switch between growth and quiescence. PMID- 11024050 TI - Transcriptional activation of the cyclin D1 gene is mediated by multiple cis elements, including SP1 sites and a cAMP-responsive element in vascular endothelial cells. AB - In an attempt to examine the mechanisms by which transcriptional activity of the cyclin D1 promoter is regulated in vascular endothelial cells (EC), we examined the cis-elements in the human cyclin D1 promoter, which are required for transcriptional activation of the gene. The results of luciferase assays showed that transcriptional activity of the cyclin D1 promoter was largely mediated by SP1 sites and a cAMP-responsive element (CRE). DNA binding activity at the SP1 sites, which was analyzed by electrophoretic mobility shift assays, was significantly increased in the early to mid G(1) phase, whereas DNA binding activity at CRE did not change significantly. Furthermore, Induction of the cyclin D1 promoter activity in the early to mid G(1) phase depended largely on the promoter fragment containing the SP1 sites, whereas the proximal fragment containing CRE but not the SP1 sites was constitutively active. Finally, the increase in DNA binding and promoter activities via the SP1 sites was mediated by the Ras-dependent pathway. The results suggested that the activation of the cyclin D1 gene in vascular ECs was regulated by a dual system; one was inducible in the G(1) phase, and the other was constitutively active. PMID- 11024051 TI - Sds3 (suppressor of defective silencing 3) is an integral component of the yeast Sin3[middle dot]Rpd3 histone deacetylase complex and is required for histone deacetylase activity. AB - SDS3 (suppressor of defective silencing 3) was originally identified in a screen for mutations that cause increased silencing of a crippled HMR silencer in a rap1 mutant background. In addition, sds3 mutants have phenotypes very similar to those seen in sin3 and rpd3 mutants, suggesting that it functions in the same genetic pathway. In this manuscript we demonstrate that Sds3p is an integral subunit of a previously identified high molecular weight Rpd3p.Sin3p containing yeast histone deacetylase complex. By analyzing an sds3Delta strain we show that, in the absence of Sds3p, Sin3p can be chromatographically separated from Rpd3p, indicating that Sds3p promotes the integrity of the complex. Moreover, the remaining Rpd3p complex in the sds3Delta strain had little or no histone deacetylase activity. Thus, Sds3p plays important roles in the integrity and catalytic activity of the Rpd3p.Sin3p complex. PMID- 11024052 TI - Transcription of the interferon gamma (IFN-gamma )-inducible chemokine Mig in IFN gamma-deficient mice. AB - MuMig or Mig (murine monokine induced by interferon gamma) is a CXC chemokine whose induction is thought to be strictly dependent on interferon gamma (IFN gamma). Here we have studied the expression of this chemokine gene in various organs of mice infected with vaccinia virus. We have employed animals deficient in either IFN-gamma (IFN-gamma(-/-)), or receptors for IFN-alpha/beta, IFN-gamma, or both IFN-alpha/beta and IFN-gamma (DR(-/-)) to dissect out the role of interferons in the induction of Mig during the host response to virus infection. Our data show that Mig mRNA and protein are expressed in organs of vaccinia virus infected IFN-gamma(-/-) mice, albeit at lower levels compared with infected, wild type animals. In the DR(-/-) mice and in IFN-gamma(-/-) mice treated with a neutralizing antibody to IFN-alpha/beta, Mig mRNA transcripts were completely absent. Our data indicate that, in vaccinia virus-infected IFN-gamma(-/-) mice, Mig mRNA expression is mediated through the interaction between IFN-gamma responsive element 1 (gammaRE-1) and IFN-alpha/beta-induced STAT-1 complex referred to as IFN-gamma response factor 2 (gammaRF-2). Further, our findings support the view that gammaRF-2 is the IFN-alpha/beta induced STAT-1 complex, IFN alpha-activated factor. We have found that, in the absence of IFN-gamma, IFN alpha/beta are able to induce Mig in response to a viral infection in vivo. PMID- 11024054 TI - The ATPase domain of hsp70 possesses a unique binding specificity for 3' sulfogalactolipids. AB - The region(s) of hsp70 critical for sulfogalactolipid (SGL) recognition has been defined through deletion analysis and site-directed mutagenesis. Truncated polymerase chain reaction products of hsp70 generated N-terminal fragments of 43, 35, 29, and 22 kDa. The C terminus substrate-binding domain (28 kDa) was also expressed. The N-terminal ATPase domain (rP43) shared the binding specificity of hsp70, because only sulfogalactosyl ceramide and sulfogalactosyl glycerolipid were recognized by both TLC overlay and RELISA. The C-terminal domain showed no binding. SGL binding of rP29 and rP22 was severely reduced. The loss of SGL binding for rP35 by RELISA but not TLC overlay was considered as a function of receptor presentation. The truncation of rP43 to rP35 demonstrates that residues 318-387 (the base of the ATP binding cleft) are critical for high affinity SGL binding. Mutagenesis showed that Arg(342) and Phe(198) are crucial for this process. SGL binding, mediated by these conserved residues within the ATPase domain of hsp70, implies that this binding specificity is evolutionarily conserved. PMID- 11024053 TI - The addition of bisecting N-acetylglucosamine residues to E-cadherin down regulates the tyrosine phosphorylation of beta-catenin. AB - The enzyme GnT-III (beta 1,4-N-acetylglucosaminyltransferase III) catalyzes the addition of a bisecting N-acetylglucosamine (GlcNAc) residue on glycoproteins. Our previous study described that the transfection of GnT-lll into mouse melanoma cells results in the enhanced expression of E-cadherin, which in turn leads to the suppression of lung metastasis. It has recently been proposed that the phosphorylation of a tyrosine residue of beta-catenin is associated with cell migration. The present study reports on the importance of bisecting GlcNAc residues by GnT-lll on tyrosine phosphorylation of beta-catenin using three types of cancer cell lines. An addition of bisecting GlcNAc residues to E-cadherin leads to an alteration in cell morphology and the localization of beta-catenin after epidermal growth factor stimulation. These changes are the result of a down regulation in the tyrosine phosphorylation of beta-catenin. In addition, tyrosine phosphorylation of beta-catenin by transfection of constitutively active c-src was suppressed in GnT-III transfectants as well as in the case of epidermal growth factor stimulation. Treatment with tunicamycin abolished any differences in beta-catenin phosphorylation for the mock vis a vis the GnT-lll transfectants. Thus, the addition of a specific N-glycan structure, the bisecting GlcNAc to E cadherin-beta-catenin complex, down-regulates the intracellular signaling pathway, suggesting its implication in cell motility and the suppression of cancer metastasis. PMID- 11024055 TI - Molecular determinants of voltage-dependent gating and binding of pore-blocking drugs in transmembrane segment IIIS6 of the Na(+) channel alpha subunit. AB - Mutations of amino acid residues in the inner two-thirds of the S6 segment in domain III of the rat brain type IIA Na(+) channel (G1460A to I1473A) caused periodic positive and negative shifts in the voltage dependence of activation, consistent with an alpha-helix having one face on which mutations to alanine oppose activation. Mutations in the outer one-third of the IIIS6 segment all favored activation. Mutations in the inner half of IIIS6 had strong effects on the voltage dependence of inactivation from closed states without effect on open state inactivation. Only three mutations had strong effects on block by local anesthetics and anticonvulsants. Mutations L1465A and I1469A decreased affinity of inactivated Na(+) channels up to 8-fold for the anticonvulsant lamotrigine and its congeners 227c89, 4030w92, and 619c89 as well as for the local anesthetic etidocaine. N1466A decreased affinity of inactivated Na(+) channels for the anticonvulsant 4030w92 and etidocaine by 3- and 8-fold, respectively, but had no effect on affinity of the other tested compounds. Leu-1465, Asn-1466, and Ile 1469 are located on one side of the IIIS6 helix, and mutation of each caused a positive shift in the voltage dependence of activation. Evidently, these amino acid residues face the lumen of the pore, contribute to formation of the high affinity receptor site for pore-blocking drugs, and are involved in voltage dependent activation and coupling to closed-state inactivation. PMID- 11024056 TI - DNA binding and protein-protein interaction sites in MutS, a mismatched DNA recognition protein from Thermus thermophilus HB8. AB - The mismatch repair system repairs mismatched base pairs, which are caused by either DNA replication errors, DNA damage, or genetic recombination. Mismatch repair begins with the recognition of mismatched base pairs in DNA by MutS. Protein denaturation and limited proteolysis experiments suggest that Thermus thermophilus MutS can be divided into three structural domains as follows: A (N terminal domain), B (central domain), and C (C-terminal domain) (Tachiki, H., Kato, R., Masui, R., Hasegawa, K., Itakura, H., Fukuyama, K., and Kuramitsu, S. (1998) Nucleic Acids Res. 26, 4153-4159). To investigate the functions of each domain in detail, truncated genes corresponding to the domains were designed. The gene products were overproduced in Escherichia coli, purified, and assayed for various activities. The MutS-MutS protein interaction site was determined by size exclusion chromatography to be located in the B domain. The B domain was also found to possess nonspecific double-stranded DNA-binding ability. The C domain, which contains a Walker's A-type nucleotide-binding motif, demonstrated ATPase activity and specific DNA recognition of mismatched base pairs. These ATPase and specific DNA binding activities were found to be dependent upon C domain dimerization. PMID- 11024057 TI - The role of histidine 632 in catalysis by human topoisomerase I. AB - Based on the crystal structure of human topoisomerase I, we hypothesized that hydrogen bonding between the side chain of the highly conserved His(632) and one of the nonbridging oxygens of the scissile phosphate contributes to catalysis by stabilizing the transition state. This hypothesis has been tested by examining the effects of changing His(632) to glutamine, asparagine, alanine, and tryptophan. The change to glutamine reduced both the relaxation activity and single-turnover cleavage activity by approximately 100-fold, whereas the same change at three other conserved histidines (positions 222, 367, and 406) had no significant effect on the relaxation activity. The properties of the mutant protein containing asparagine instead of histidine at position 632 were similar to those of the glutamine mutant, whereas mutations to alanine or tryptophan reduced the activity by approximately 4 orders of magnitude. The reduction in activity for the mutants was not due to alterations in substrate binding affinities or changes in the cleavage specificities of the proteins. The above results for the glutamine mutation in conjunction with the similar effects of pH on the wild type and the H632Q mutant enzyme rule out the possibility that His(632) acts as a general acid to protonate the leaving 5'-oxygen during the cleavage reaction. Taken together, these data strongly support the hypothesis that the only role for His(632) is to stabilize the pentavalent transition state through hydrogen bonding to one of the nonbridging oxygens. PMID- 11024058 TI - Packing-induced conformational and functional changes in the subunits of alpha crystallin. AB - The heteroaggregate alpha-crystallin and homoaggregates of its subunits, alphaA- and alphaB-crystallins, function like molecular chaperones and prevent the aggregation of several proteins. Although modulation of the chaperone-like activity of alpha-crystallin by both temperature and chaotropic agents has been demonstrated in vitro, the mechanism(s) of its regulation in vivo have not been elucidated. The subunits of alpha-crystallin exchange freely, resulting in its dynamic and variable quaternary structure. Mixed aggregates of the alpha crystallins and other mammalian small heat shock proteins (sHSPs) have also been observed in vivo. We have investigated the time-dependent structural and functional changes during the course of heteroaggregate formation by the exchange of subunits between homoaggregates of alphaA- and alphaB-crystallins. Native isoelectric focusing was used to follow the time course of subunit exchange. Circular dichroism revealed large tertiary structural alterations in the subunits upon subunit exchange and packing into heteroaggregates, indicating specific homologous and heterologous interactions between the subunits. Subunit exchange also resulted in quaternary structural changes as demonstrated by gel filtration chromatography. Interestingly, we found time-dependent changes in chaperone-like activity against the dithiothreitol-induced aggregation of insulin, which correlated with subunit exchange and the resulting tertiary and quaternary structural changes. Heteroaggregates of varying subunit composition, as observed during eye lens epithelial cell differentiation, generated by subunit exchange displayed differential chaperone-like activity. It was possible to alter chaperone-like activity of preexisting oligomeric sHSPs by alteration of subunit composition by subunit exchange. Our results demonstrate that subunit exchange and the resulting structural and functional changes observed could constitute a mechanism of regulation of chaperone-like activity of alpha-crystallin (and possibly other mammalian sHSPs) in vivo. PMID- 11024059 TI - Role of exon 2-encoded beta -domain of the von Hippel-Lindau tumor suppressor protein. AB - Sporadic clear cell renal carcinomas frequently harbor inactivating mutations in exon 2 of the von Hippel-Lindau (VHL) tumor suppressor gene. Here, we examine the effect of the loss of exon 2-encoded beta-domain function on VHL biochemical properties. Exon 2-encoded residues are required for VHL-mediated NEDD8 conjugation on cullin-2 and assembly with hypoxia-inducible factor alpha (HIFalpha) and fibronectin. These residues are not essential for VHL ability to assemble with elongin BC/cullin-2, to display E3 ubiquitin ligase activity in vitro and to confer energy-dependent nuclear import properties to a reporter protein. Localization studies in HIF-1alpha-null embryonic cells suggest that exon 2-encoded beta-domain mediates transcription-dependent nuclear/cytoplasmic shuttling of VHL independently of assembly with HIF-1alpha and oxygen concentration. Exon 3-encoded alpha-helical domain is required for VHL complex formation with BC/cullin-2 and E3 ubiquitin ligase activity, for binding to HIFalpha/fibronectin, but this domain is not essential for transcription dependent nuclear/cytoplasmic trafficking. VHL(-/-) renal carcinoma cells expressing beta-domain mutants failed to produce an extracellular fibronectin matrix and to degrade HIFalpha, which accumulated exclusively in the nucleus of normoxic cells. These results demonstrate that exon 2-encoded residues are involved in two independent functions: substrate protein recognition and transcription-dependent nuclear/cytoplasmic trafficking. They also suggest that beta-domain mutations inactivate VHL function differently than alpha-domain mutations, potentially providing an explanation for the relationship between different mutations of the VHL gene and clinical outcome. PMID- 11024061 TI - Revisiting the catalytic CuZ cluster of nitrous oxide (N2O) reductase. Evidence of a bridging inorganic sulfur. AB - Nitrous-oxide reductases (N2OR) catalyze the two-electron reduction of N(2)O to N(2). The crystal structure of N2ORs from Pseudomonas nautica (Pn) and Paracoccus denitrificans (Pd) were solved at resolutions of 2.4 and 1.6 A, respectively. The Pn N2OR structure revealed that the catalytic CuZ center belongs to a new type of metal cluster in which four copper ions are liganded by seven histidine residues. A bridging oxygen moiety and two other hydroxide ligands were proposed to complete the ligation scheme (Brown, K., Tegoni, M., Prudencio, M., Pereira, A. S., Besson, S., Moura, J. J. G., Moura, I., and Cambillau, C. (2000) Nat. Struct. Biol. 7, 191-195). However, in the CuZ cluster, inorganic sulfur chemical determination and the high resolution structure of Pd N2OR identified a bridging inorganic sulfur instead of an oxygen. This result reconciles the novel CuZ cluster with the hitherto puzzling spectroscopic data. PMID- 11024060 TI - Mutations in the Kv beta 2 binding site for NADPH and their effects on Kv1.4. AB - Kv beta 2 enhances the rate of inactivation and level of expression of Kv1.4 currents. The crystal structure of Kv beta 2 binds NADP(+), and it has been suggested that Kv beta 2 is an oxidoreductase enzyme (). To investigate how this function might relate to channel modulation, we made point mutations in Kv beta 2 in either the NADPH docking or putative catalytic sites. Using the yeast two hybrid system, we found that these mutations did not disrupt the interaction of Kv beta 2 with Kv alpha 1 channels. To characterize the Kv beta 2 mutants functionally, we coinjected wild-type or mutant Kv beta 2 cRNAs and Kv1.4 cRNA in Xenopus laevis oocytes. Kv beta 2 increased both the amplitude and rate of inactivation of Kv1.4 currents. The cellular content of Kv1.4 protein was unchanged on Western blot, but the amount in the plasmalemma was increased. Mutations in either the orientation or putative catalytic sites for NADPH abolished the expression-enhancing effect on Kv1.4 current. Western blots showed that both types of mutation reduced Kv1.4 protein. Like the wild-type Kv beta 2, both types of mutation increased the rate of inactivation of Kv1.4, confirming the physical association of mutant Kv beta 2 subunits with Kv1.4. Thus, mutations that should interfere with NADPH function uncouple the expression-enhancing effect of Kv beta 2 on Kv1.4 currents from its effect on the rate of inactivation. These results suggest that the binding of NADPH and the putative oxidoreductase activity of Kv beta 2 may play a role in the processing of Kv1.4. PMID- 11024062 TI - Pharmacology of GABA(A) receptors of retinal dopaminergic neurons. AB - When the vertebrate retina is stimulated by light, a class of amacrine or interplexiform cells release dopamine, a modulator responsible for neural adaptation to light. In the intact retina, dopamine release can be pharmacologically manipulated with agonists and antagonists at GABA(A) receptors, and dopaminergic (DA) cells receive input from GABAergic amacrines. Because there are only 450 DA cells in each mouse retina and they cannot be distinguished in the living state from other cells on the basis of their morphology, we used transgenic technology to label DA cells with human placental alkaline phosphatase, an enzyme that resides on the outer surface of the cell membrane. We could therefore identify DA cells in vitro after dissociation of the retina and investigate their activity with whole cell voltage clamp. We describe here the pharmacological properties of the GABA(A) receptors of solitary DA cells. GABA application induces a large inward current carried by chloride ions. The receptors are of the GABA(A) type because the GABA-evoked current is blocked by bicuculline. Their affinity for GABA is very high with an EC(50) value of 7.4 microM. Co-application of benzodiazepine receptor ligands causes a strong increase in the peak current induced by GABA (maximal enhancement: CL-218872 220%; flunitrazepam 214%; zolpidem 348%) proving that DA cells express a type I benzodiazepine-receptor (BZ1). GABA-evoked currents are inhibited by Zn(2+) with an IC(50) of 58.9 +/- 8.9 microM. Furthermore, these receptors are strongly potentiated by the modulator alphaxalone with an EC(50) of 340 +/- 4 nM. The allosteric modulator loreclezole increases GABA receptor currents by 43% (1 microM) and by 107% (10 microM). Using outside-out patches, we measured in single channel recordings a main conductance (29 pS) and two subconductance (20 and 9 pS) states. We have previously shown by single-cell RT-PCR and immunocytochemistry that DA cells express seven different GABA(A) receptor subunits (alpha1, alpha3, alpha4, beta1, beta3, gamma1, gamma2(S), and gamma2(L)) and by immunocytochemistry that all subunits are expressed in the intact retina. We show here that at least alpha1, beta3 and gamma2 subunits are assembled into functional receptors. PMID- 11024063 TI - Intermittency in the control of continuous force production. AB - The purpose of the current investigation was to examine the influence of intermittency in visual information processes on intermittency in the control continuous force production. Adult human participants were required to maintain force at, and minimize variability around, a force target over an extended duration (15 s), while the intermittency of on-line visual feedback presentation was varied across conditions. This was accomplished by varying the frequency of successive force-feedback deliveries presented on a video display. As a function of a 128-fold increase in feedback frequency (0.2 to 25.6 Hz), performance quality improved according to hyperbolic functions (e.g., force variability decayed), reaching asymptotic values near the 6.4-Hz feedback frequency level. Thus, the briefest interval over which visual information could be integrated and used to correct errors in motor output was approximately 150 ms. The observed reductions in force variability were correlated with parallel declines in spectral power at about 1 Hz in the frequency profile of force output. In contrast, power at higher frequencies in the force output spectrum were uncorrelated with increases in feedback frequency. Thus, there was a considerable lag between the generation of motor output corrections (1 Hz) and the processing of visual feedback information (6.4 Hz). To reconcile these differences in visual and motor processing times, we proposed a model where error information is accumulated by visual information processes at a maximum frequency of 6.4 per second, and the motor system generates a correction on the basis of the accumulated information at the end of each 1-s interval. PMID- 11024064 TI - Serotonergic Raphe magnus cells that respond to noxious tail heat are not ON or OFF cells. AB - Pharmacological studies have suggested that serotonergic cells in RM contribute to both the inhibition and facilitation of spinal nociceptive transmission. Physiological studies in the medullary nucleus raphe magnus (RM) and adjacent nucleus reticularis magnocellularis have identified putative nociceptive inhibitory OFF cells and nociceptive-facilitatory neurons ON cells by their responses to noxious thermal stimulation. The present study was designed to determine 1) whether any serotonergic RM cells respond to noxious thermal stimulation and 2) whether noxious heat-responsive serotonergic cells should be classified as ON or OFF cells. Serotonergic cells (n = 150) were identified by physiological criteria in anesthetized rats; 30 of 32 cells tested contained serotonin immunoreactivity. Noxious tail heat elicited a neuronal response in less than a quarter of the serotonergic cells. Most serotonergic cells that responded to tail heat were excited (n = 25), while a small minority of the cells tested were inhibited (n = 8). The tail heat-evoked responses of serotonergic cells were small in magnitude, averaging five to eight spikes in 10 s. Excitatory responses rarely persisted for more than 10 s, while inhibitory responses rarely persisted for more than 20 s. The tail heat-evoked responses of serotonergic cells were compared to those of non-serotonergic cells (n = 186). Non serotonergic cells that responded to noxious tail heat had significantly greater response magnitudes, averaging 75-95 spikes in 10 s, than heat-responsive serotonergic cells. In addition, most heat-responsive non-serotonergic cells responded for at least 30 s after stimulus onset. These results demonstrate that the tail heat-evoked responses of serotonergic RM cells are qualitatively and quantitatively distinct from those of non-serotonergic ON and OFF cells. It is therefore unlikely that serotonergic RM cells, even the subpopulation that responds to noxious tail heat, share a physiological function with ON and OFF cells. PMID- 11024065 TI - Development of spontaneous glycinergic currents in the Mauthner neuron of the zebrafish embryo. AB - We used whole cell and outside-out patch-clamp techniques with reticulospinal Mauthner neurons of zebrafish embryos to investigate the developmental changes in the properties of glycinergic synaptic currents in vivo from the onset of synaptogenesis. Miniature inhibitory postsynaptic currents (mIPSCs) were isolated and recorded in the presence of TTX (1 microM), kynurenic acid (1 mM), and bicuculline (10 microM) and were found to be sensitive to strychnine (1 microM). The mIPSCs were first observed in 26-29 h postfertilization (hpf) embryos at a very low frequency of approximately 0.04 Hz, which increased to approximately 0.5 Hz by 30-40 hpf, and was approximately 10 Hz in newly hatched (>50 hpf) larvae, indicating an accelerated increase in synaptic activity. At all embryonic stages, the amplitudes of the mIPSCs were variable but their means were similar ( approximately 100 pA), suggesting rapid formation of the postsynaptic matrix. The 20-80% rise times of mIPSCs in embryos were longer (0.6-1.2 ms) than in larvae (approximately 0.3 ms), likely due to slower diffusion of glycine at the younger, immature synapses. The mIPSCs decayed with biexponential (tau(off1) and tau(off2)) time courses with a half-width in 26-29 hpf embryos that was longer and more variable than in older embryos and larvae. In 26- to 29-hpf embryos, tau(off1) was approximately 15 ms and tau(off2) was approximately 60 ms, representing events of intermediate duration; but occasionally long mIPSCs were observed in some cells where tau(off1) was approximately 40 ms and tau(off2) was approximately 160 ms. In 30-40 hpf embryos, the events were faster, with tau(off1) approximately 9 ms and tau(off2) approximately 40 ms, and in larvae, events declined somewhat further to tau(off1) approximately 4 ms and tau(off2) approximately 30 ms. Point-per-point amplitude histograms of the decay of synaptic events at all stages resulted in the detection of similar single channel conductances estimated as approximately 45 pS, indicating the presence of heteromeric glycine receptors (GlyRs) from the onset of synaptogenesis. Fast-flow (1 ms) application of a saturating concentration of glycine (3-10 mM) to outside out patches obtained at 26-29 hpf revealed GlyR currents that decayed biexponentially with time constants resembling the values found for intermediate and long mIPSCs; by 30-40 hpf, the GlyR currents resembled fast mIPSCs. These observations indicate that channel kinetics limited the mIPSC duration. Our data suggest that glycinergic mIPSCs result from the activation of a mixture of fast and slow GlyR subtypes, the properties and proportion of which determine the decay of the synaptic events in the embryos. PMID- 11024066 TI - Influence of leg muscle vibration on human walking. AB - We studied the effect of vibratory stimulation of different leg muscles [bilateral quadriceps (Q), hamstring (HS) muscles, triceps surae (TS), and tibialis anterior (TA)] in seven normal subjects during 1) quiet standing, 2) stepping in place movements, and 3) walking on the treadmill. The experiments were performed in a dimly illuminated room, and the subjects were given the instruction not to resist the applied perturbation. In one condition the velocity of the treadmill was controlled by a feedback from the subject's current position. In normal standing, TA vibration elicited a prominent forward body tilt, whereas HS and TS vibration elicited backward trunk or whole body inclination, respectively. Q vibration had little effect. During stepping in place, continuous HS vibration produced an involuntary forward stepping at about 0.3 m s(-1) without modifying the stepping frequency. When the subjects (with eyes closed) kept a hand contact with an external still object, they did not move forward but perceived an illusory forward leg flexion relative to the trunk. Q, TS, and TA vibration did not cause any systematic body translation nor illusory changes in body configuration. In treadmill locomotion, HS vibration produced an involuntary steplike increase of walking speed (by 0.1-0.6 m.s(-1)). Continuous vibration elicited larger speed increments than phasic stimulation during swing or stance phase. For phasic stimulation, HS vibration tended to be more effective when applied during swing than during stance phase. Q, TA, and TS vibration had little if any effect. Vibration of thigh muscles altered the walking speed depending on the direction of progression. During backward locomotion, the walking speed tended to decrease after HS vibration, whereas it significantly increased after Q vibration. Thus the influence of leg muscle vibration on stepping in place and locomotion differed significantly from that on normal posture. We suggest that the proprioceptive input from thigh muscles may convey information about the velocity of the foot movement relative to the trunk. PMID- 11024067 TI - Context-dependent smooth eye movements evoked by stationary visual stimuli in trained monkeys. AB - The appearance of a stationary but irrelevant cue triggers a smooth eye movement away from the position of the cue in monkeys that have been trained extensively to smoothly track the motion of moving targets while not making saccades to the stationary cue. We have analyzed the parameters that regulate the size of the cue evoked smooth eye movement and examined whether presentation of the cue changes the initiation of pursuit for subsequent steps of target velocity. Cues evoked smooth eye movements in blocks of target motions that required smooth pursuit to moving targets, but evoked much smaller smooth eye movements in blocks that required saccades to stationary targets. The direction of the cue-evoked eye movement was always opposite to the position of the cue and did not depend on whether subsequent target motion was toward or away from the position of fixation. The latency of the cue-evoked smooth eye movement was near 100 ms and was slightly longer than the latency of pursuit for target motion away from the position of fixation. The size of the cue-evoked smooth eye movement was as large as 10 degrees /s and decreased as functions of the eccentricity of the cue and the illumination of the experimental room. To study the initiation of pursuit in the wake of the cues, we used bilateral cues at equal eccentricities to the right and left of the position of fixation. These evoked smaller eye velocities that were consistent with vector averaging of the responses to each cue. In the wake of bilateral cues, the initiation of pursuit was enhanced for target motion away from the position of fixation, but not for target motion toward the position of fixation. We suggest that the cue-evoked smooth eye movement is related to a previously postulated on-line gain control for pursuit, and that it is a side effect of sudden activation of the gain-controlling element. PMID- 11024068 TI - Control of membrane sealing in injured mammalian spinal cord axons. AB - The process of sealing of damaged axons was examined in isolated strips of white matter from guinea pig spinal cord by recording the "compound membrane potential," using a sucrose-gap technique, and by examining uptake of horseradish peroxidase (HRP). Following axonal transection, exponential recovery of membrane potential occurred with a time constant of 20 +/- 5 min, at 37 degrees C, and extracellular calcium activity ([Ca(2+)](o)) of 2 mM. Most axons excluded HRP by 30 min following transection. The rate of sealing was reduced by lowering calcium and was effectively blocked at [Ca(2+)](o) 20 min) depotentiation, to the baseline level, on field potentials in four recording cases. The autocorrelation functions indicated that the isolated unit discharges were comparable to those of the putative DG granule cells and interneurons, shown in previous studies. The results suggest that changes in synaptic efficacy following HFS or LFS produce rather dynamic changes in cell activity in the DG. PMID- 11024083 TI - BOLD fMRI response of early visual areas to perceived contrast in human amblyopia. AB - In this study, we used a temporal two-alternative forced choice psychophysical procedure to measure the observer's perception of a 22% physical contrast grating for each eye as a function of spatial frequency in four subjects with unilateral amblyopia and in six subjects with normal vision. Contrast thresholds were also measured using a standard staircase method. Additionally, blood-oxygenation-level dependent (BOLD) functional magnetic resonance imaging (fMRI) was used to measure the neuronal response within early visual cortical areas to monocular presentations of the same 22% physical contrast gratings as a function of spatial frequency. For all six subjects with normal vision and for three subjects with amblyopia, the psychophysically measured perception of 22% contrast as a function of spatial frequency was the same for both eyes. Threshold contrast, however, was elevated for the amblyopic eye for all subjects, as expected. The magnitude of the fMRI response to 22% physical contrast within "activated" voxels was the same for each eye as a function of spatial frequency, regardless of the presence of amblyopia. However, there were always fewer "activated" fMRI voxels during amblyopic stimulation than during normal eye stimulation. These results are consistent with the hypotheses that contrast thresholds are elevated in amblyopia because fewer neurons are responsive during amblyopic stimulation, and that the average firing rate of the responsive neurons, which reflects the perception of contrast, is unaffected in amblyopia. PMID- 11024084 TI - Dendritic calcium accumulation associated with direction-selective adaptation in visual motion-sensitive neurons in vivo. AB - Motion adaptation in directionally selective tangential cells (TC) of the fly visual system has previously been explained as a presynaptic mechanism. Based on the observation that adaptation is in part direction selective, which is not accounted for by the former models of motion adaptation, we investigated whether physiological changes located in the TC dendrite can contribute to motion adaptation. Visual motion in the neuron's preferred direction (PD) induced stronger adaptation than motion in the opposite direction and was followed by an afterhyperpolarization (AHP). The AHP subsides in the same time as adaptation recovers. By combining in vivo calcium fluorescence imaging with intracellular recording, we show that dendritic calcium accumulation following motion in the PD is correlated with the AHP. These results are consistent with a calcium-dependent physiological change in TCs underlying adaptation during continuous stimulation with PD motion, expressing itself as an AHP after the stimulus stops. However, direction selectivity of adaptation is probably not solely related to a calcium dependent mechanism because direction-selective effects can also be observed for fast moving stimuli, which do not induce sizeable calcium accumulation. In addition, a comparison of two classes of TCs revealed differences in the relationship of calcium accumulation and AHP when the stimulus velocity was varied. Thus the potential role of calcium in motion adaptation depends on stimulation parameters and cell class. PMID- 11024085 TI - Sensory fibers of the pelvic nerve innervating the Rat's urinary bladder. AB - Much attention has been given to the pelvic nerve afferent innervation of the urinary bladder; however, reports differ considerably in descriptions of afferent receptor types, their conduction velocities, and their potential roles in bladder reflexes and sensation. The present study was undertaken to do a relatively unbiased sampling of bladder afferent fibers of the pelvic nerve in adult female rats. The search stimulus for units to be studied was electrical stimulation of both the bladder nerves and the pelvic nerve. Single-unit activity of 100 L(6) dorsal root fibers, activated by both pelvic and bladder nerve stimulation, was analyzed. Sixty-five units had C-fiber and 35 units had Adelta-fiber conduction velocities. Receptive characteristics were established by direct mechanical stimulation, filling of the bladder with 0.9% NaCl at a physiological speed and by filling the bladder with solutions containing capsaicin, potassium, or turpentine oil. The majority (61) of these fibers were unambiguously excited by bladder filling with 0.9% NaCl and were classified as mechanoreceptors. All mechanoreceptors with receptive fields on the body of the bladder had low pressure thresholds ( or =98.7% homology and conservation of structurally important domains. Chimeras between the R5X4 89.6 prototype and an R5 variant indicated that multiple regions contributed to the use of CXCR4, while chimeras with the X4 variant implicated a single residue in V4 in CCR5 use. These results confirm, at the molecular level, both that dualtropic variants are a predominant component of late-stage syncytium-inducing isolates and that variants restricted to each coreceptor coexist with dualtropic species in vivo. Coreceptor-restricted minority variants may reflect residual R5 species from earlier in disease as well as emerging X4 variants. PMID- 11024155 TI - Sequestration of TT virus of restricted genotypes in peripheral blood mononuclear cells. AB - Peripheral blood mononuclear cells (PBMC) harbored TT virus (TTV) of genotypes (3 and 4) different from those (1 and 2) of free virions in plasma of the same individuals. PBMC may act as a reservoir, and TTV of particular genotypes might have tropism for hematopoietic cells. PMID- 11024156 TI - HLA class I-restricted cytotoxic T-cell epitopes of the respiratory syncytial virus fusion protein. AB - Virus-specific cytotoxic T lymphocytes (CTL) play a major role in the clearance of respiratory syncytial virus (RSV) infection. We have generated cytotoxic T cell clones (TCC) from two infants who had just recovered from severe RSV infection. These TCC were functionally characterized and used to identify HLA class I (B57 and C12)-restricted CTL epitopes of RSV. PMID- 11024157 TI - Identification of upregulated genes in scrapie-infected brain tissue. AB - The pathogenesis of scrapie, and of neurodegenerative diseases in general, is still insufficiently understood and is therefore being intensely researched. There is abundant evidence that the activation of glial cells precedes neurodegeneration and may thus play an important role in disease development and progression. The identification of genes with altered expression patterns in the diseased brain may provide insight on the molecular level into the process which ultimately leads to neuronal loss. Differentially expressed genes in scrapie infected brain tissue were enriched by the suppression subtractive hybridization technique, molecularly cloned, and further characterized. Northern blotting and nucleotide sequencing confirmed the identities of 19 upregulated genes, 11 of which were unknown to be affected by scrapie. A considerable number of these 19 genes, namely those encoding interferon-inducible protein 10 (IP-10), 2',5' oligo(A) synthetase, Mx protein, IIGP protein, major histocompatibility complex classes I and II, complement, and beta(2)-microglobulin, were inducible by interferons (IFNs), suggesting that an IFN response is a possible mechanism of gene activation in scrapie. Among the newly found genes, that coding for 2',5' oligo(A) synthetase is of special interest because it could contribute to the apoptotic loss of neuronal cells via RNase L activation. In addition, upregulation of the chemokine IP-10 and B-lymphocyte chemoattractant mRNAs was seen at relatively early stages of the disease and was sustained throughout disease development. PMID- 11024158 TI - A significant number of human immunodeficiency virus epitope-specific cytotoxic T lymphocytes detected by tetramer binding do not produce gamma interferon. AB - Despite the seemingly important role of cytotoxic T-lymphocyte (CTL) responses in human immunodeficiency virus (HIV) disease pathogenesis, their measurement has relied on a variety of different techniques. We utilized three separate methodologies for the detection of CTLs in a cohort of HIV-infected individuals who were also human leukocyte antigen A2 (HLA-A2) positive. Among the different CTL assays, a correlation was seen only when the Gag epitope-specific HLA A*0201 restricted tetramer assay was compared with the ELISPOT assay performed after stimulation with the Gag epitope; however, this correlation was of borderline statistical significance. On average, the tetramer reagent detected a 10-fold higher number of cells than were seen to produce gamma interferon by the ELISPOT assay. The implications of this CTL assay comparison and the possibility of phenotypic differences in HIV-specific CD8(+) T lymphocytes are discussed. PMID- 11024159 TI - Incorporation of HLA-DR into the envelope of human immunodeficiency virus type 1 in vivo: correlation with stage of disease and presence of opportunistic infection. AB - Human immunodeficiency virus type 1 (HIV-1) bearing HLA-DR in its envelope was detected in plasma from all patients with chronic HIV-1 infection (n = 16) and was present at higher levels in patients with active tuberculosis coinfection (n = 6). Intriguingly, however, HLA-DR was not detectable in HIV-1 from patients during primary viremia (n = 6), suggesting the possibility of virus replication in less-activated cells. PMID- 11024160 TI - Role of the Rous sarcoma virus p10 domain in shape determination of gag virus like particles assembled in vitro and within Escherichia coli. AB - Purified retrovirus Gag proteins can assemble in vitro into virus-like particles (VLPs) in the presence of RNA. It was shown previously that a Rous sarcoma virus Gag protein missing only the protease domain forms spherical particles resembling immature virions lacking a membrane but that a similar protein missing the p10 domain forms tubular particles. Thus, p10 plays a role in spherical particle formation. To further study this shape-determining function, we dissected the p10 domain by mutagenesis and examined VLPs assembled within Escherichia coli or assembled in vitro from purified proteins. The results identified a minimal contiguous segment of 25 amino acid residues at the C terminus of p10 that is sufficient to restore efficient spherical assembly to a p10 deletion mutant. Random and site-directed mutations were introduced into this segment of polypeptide, and the shapes of particles formed in E. coli were examined in crude extracts by electron microscopy. Three phenotypes were observed: tubular morphology, spherical morphology, or no regular structure. While the particle morphology visualized in crude extracts generally was the same as that visualized for purified proteins, some tubular mutants scored as spherical when tested as purified proteins, suggesting that a cellular factor may also play a role in shape determination. We also examined the assembly properties of smaller Gag proteins consisting of the capsid protein-nucleocapsid protein (CA-NC) domains with short N-terminal extensions or deletions. Addition of one or three residues allowed CA-NC to form spheres instead of tubes in vitro, but the efficiency of assembly was extremely low. Deletion of the N-terminal residue(s) abrogated assembly. Taken together, these results imply that the N terminus of CA and the adjacent upstream 25 residues play an important role in the polymerization of the Gag protein. PMID- 11024161 TI - Comparative analysis of secondary structure of insect mitochondrial small subunit ribosomal RNA using maximum weighted matching. AB - Comparative analysis is the preferred method of inferring RNA secondary structure, but its use requires considerable expertise and manual effort. As the importance of secondary structure for accurate sequence alignment and phylogenetic analysis becomes increasingly realised, the need for secondary structure models for diverse taxonomic groups becomes more pressing. The number of available structures bears little relation to the relative diversity or importance of the different taxonomic groups. Insects, for example, comprise the largest group of animals and yet are very poorly represented in secondary structure databases. This paper explores the utility of maximum weighted matching (MWM) to help automate the process of comparative analysis by inferring secondary structure for insect mitochondrial small subunit (12S) rRNA sequences. By combining information on correlated changes in substitutions and helix dot plots, MWM can rapidly generate plausible models of secondary structure. These models can be further refined using standard comparative techniques. This paper presents a secondary structure model for insect 12S rRNA based on an alignment of 225 insect sequences and an alignment for 16 exemplar insect sequences. This alignment is used as a template for a web server that automatically generates secondary structures for insect sequences. PMID- 11024162 TI - Structure and function of the fourth subunit (Dpb4p) of DNA polymerase epsilon in Saccharomyces cerevisiae. AB - DNA polymerase epsilon (Polepsilon) of Saccharomyces cerevisiae is purified as a complex of four polypeptides with molecular masses of >250, 80, 34 (and 31) and 29 kDa as determined by SDS-PAGE. The genes POL2, DPB2 and DPB3, encoding the catalytic Pol2p, the second (Dpb2p) and the third largest subunits (Dpb3p) of the complex, respectively, were previously cloned and characterised. This paper reports the partial amino acid sequence of the fourth subunit (Dpb4p) of Polepsilon. This protein sequence matches parts of the predicted amino acid sequence from the YDR121w open reading frame on S.cerevisiae chromosome IV. Thus, YDR121w was renamed DPB4. A deletion mutant of DPB4 (Deltadpb4) is not lethal, but chromosomal DNA replication is slightly disturbed in this mutant. A double mutant haploid strain carrying the Deltadpb4 deletion and either pol2-11 or dpb11 1 is lethal at all temperatures tested. Furthermore, the restrictive temperature of double mutants carrying Deltadpb4 and dpb2-1, rad53-1 or rad53-21 is lower than in the corresponding single mutants. These results strongly suggest that Dpb4p plays an important role in maintaining the complex structure of Polepsilon in S.cerevisiae, even if it is not essential for cell growth. Structural homologues of DPB4 are present in other eukaryotic genomes, suggesting that the complex structure of S. cerevisiae Polepsilon is conserved in eukaryotes. PMID- 11024163 TI - Structure of HAP1-PC7 bound to DNA: implications for DNA recognition and allosteric effects of DNA-binding on transcriptional activation. AB - HAP1 is a transcription factor in yeast whose DNA-binding domain has been implicated in directly affecting transcriptional activation. Two separate mutations in the DNA-binding domain, S63G (HAP1-PC7) and S63R (HAP1-18), retain wild-type binding affinity. However, HAP1-PC7 is transcriptionally silent while HAP1-18 shows highly elevated levels of transcription. We have determined the X ray crystal structure of the DNA-binding domain of HAP1-PC7 bound to its DNA target, UAS(CYC7), and compared it to the previously solved HAP1-wt and HAP1-18 complexes to UAS(CYC7). Additionally, we have quantitatively compared the DNA binding affinity and specificity of the HAP1-PC7, HAP1-18 and HAP1-wt DNA-binding domains. We show that, although the DNA-binding domains of these three proteins bind UAS(CYC7) with comparable affinity and specificity, the protein-DNA interactions are dramatically different between the three complexes. Conserved protein-DNA interactions are largely restricted to an internal DNA sequence that excludes one of the two conserved DNA half-sites of UAS(CYC7) suggesting a mode of recognition distinct from other HAP1 family members. Alternative protein-DNA interactions result in divergent DNA configurations between the three complexes. These results suggest that the differential transcriptional activities of the HAP1, HAP1-18 and HAP1-PC7 proteins are due, at least in part, to alternative protein-DNA contacts, and implies that HAP1-DNA interactions have direct allosteric effects on transcriptional activation. PMID- 11024164 TI - Functional studies of the BTB domain in the Drosophila GAGA and Mod(mdg4) proteins. AB - The BTB/POZ (BTB) domain is an approximately 120 residue sequence that is conserved at the N-terminus of many proteins in both vertebrates and invertebrates. We found that the protein encoded by a lethal allele of the Drosophila modifier of mdg4 [mod(mdg4)] gene has two mutated residues in its BTB domain. The identities of the residues at the positions of these mutations are highly conserved in the BTB domain family of proteins, and when the corresponding mutations were engineered into the BTB domain-containing GAGA protein, the activity of GAGA as a transcription activator in a transient transfection assay was severely reduced. The functional equivalence of the BTB domains was established by showing that the BTB domain of the mod(mdg4) protein can effectively substitute for that of GAGA. PMID- 11024165 TI - Functional characterization of Ape1 variants identified in the human population. AB - Apurinic/apyrimidinic (AP) sites are common mutagenic and cytotoxic DNA lesions. Ape1 is the major human repair enzyme for abasic sites and incises the phosphodiester backbone 5' to the lesion to initiate a cascade of events aimed at removing the AP moiety and maintaining genetic integrity. Through resequencing of genomic DNA from 128 unrelated individuals, and searching published reports and sequence databases, seven amino acid substitution variants were identified in the repair domain of human Ape1. Functional characterization revealed that three of the variants, L104R, E126D and R237A, exhibited approximately 40-60% reductions in specific incision activity. A fourth variant, D283G, is similar to the previously characterized mutant D283A found to exhibit approximately 10% repair capacity. The most common substitution (D148E; observed at an allele frequency of 0.38) had no impact on endonuclease and DNA binding activities, nor did a G306A substitution. A G241R variant showed slightly enhanced endonuclease activity relative to wild-type. In total, four of seven substitutions in the repair domain of Ape1 imparted reduced function. These reduced function variants may represent low penetrance human polymorphisms that associate with increased disease susceptibility. PMID- 11024166 TI - Mitochondrial DNA ligase III function is independent of Xrcc1. AB - Hamster EM9 cells, which lack Xrcc1 protein, have reduced levels of DNA ligase III and are defective in nuclear base excision repair. The Xrcc1 protein stabilizes DNA ligase III and may even play a direct role in catalyzing base excision repair. Since DNA ligase III is also thought to function in mitochondrial base excision repair, it seemed likely that mitochondrial DNA ligase III function would also be dependent upon Xrcc1. However, several lines of evidence indicate that this is not the case. First, western blot analysis failed to detect Xrcc1 protein in mitochondrial extracts. Second, DNA ligase III levels present in mitochondrial protein extracts from EM9 cells were indistinguishable from those seen in similar extracts from wild-type (AA8) cells. Third, the mitochondrial DNA content of both cell lines was identical. Fourth, EM9 cells displayed no defect in their ability to repair spontaneous mitochondrial DNA damage. Fifth, while EM9 cells were far more sensitive to the cytotoxic effects of ionizing radiation due to a defect in nuclear DNA repair, there was no apparent difference in the ability of EM9 and AA8 cells to restore their mitochondrial DNA to pre-irradiation levels. Thus, mitochondrial DNA ligase III function is independent of the Xrcc1 protein. PMID- 11024167 TI - Base excision repair is efficient in cells lacking poly(ADP-ribose) polymerase 1. AB - Poly(ADP-ribose) polymerase 1 (PARP-1) is a nuclear enzyme that is activated by binding to DNA breaks induced by ionizing radiation or through repair of altered bases in DNA by base excision repair. Mice lacking PARP-1 and, in certain cases, the cells derived from these mice exhibit hypersensitivity to ionizing radiation and alkylating agents. In this study we investigated base excision repair in cells lacking PARP-1 in order to elucidate whether their augmented sensitivity to DNA damaging agents is due to an impairment of the base excision repair pathway. Extracts prepared from wild-type cells or cells lacking PARP-1 were similar in their ability to repair plasmid DNA damaged by either X-rays (single-strand DNA breaks) or by N:-methyl-N:'-nitro-N:-nitrosoguanidine (methylated bases). In addition, we demonstrated in vivo that PARP-1-deficient cells treated with N: methyl-N:'-nitro-N:-nitrosoguanidine repaired their genomic DNA as efficiently as wild-type cells. Therefore, we conclude that cells lacking PARP-1 have a normal capacity to repair single-strand DNA breaks inflicted by X-irradiation or breaks formed during the repair of modified bases. We propose that the hypersensitivity of PARP-1 null mutant cells to gamma-irradiation and alkylating agents is not directly due to a defect in DNA repair itself, but rather results from greatly reduced poly(ADP-ribose) formation during base excision repair in these cells. PMID- 11024168 TI - Localisation of the DmCdc45 DNA replication factor in the mitotic cycle and during chorion gene amplification. AB - The cdc45 protein was originally identified in Saccharomyces cerevisiae and shown to be essential for initiation of eukaryotic DNA replication. Subsequent isolation and characterisation of the corresponding genes from fission yeast, Xenopus and mammals also support a replication role for the protein in these species. They further suggest that during the course of its function cdc45 interacts with a number of other replication proteins, including minichromosome maintenance proteins, the origin recognition complex and DNA polymerase alpha. We have cloned the gene coding for cdc45 protein from Drosophila melanogaster. We have analysed the expression pattern of the cdc45 protein throughout the cell cycle and the life cycle using a combination of indirect immunofluorescence and subcellular fractionation. Our data show that cellular localisation and developmental regulation of the protein is consistent with a role in DNA replication. DmCdc45 is predominantly expressed in proliferating cells. In addition, its subcellular location is nuclear during interphase and the protein shows association with chromatin. The chromatin-associated form of the protein shows a post-translational modification, which may be involved in control of the action of the protein. DmCdc45 shows interactions with mcm proteins, however, the interactions detected show some specificity, perhaps suggesting a preferential association with particular mcm proteins. In addition we show that a stoichiometric mcm interaction may not be obligatory for the function of cdc45 in follicle cell replication, because, unlike the mcm proteins, DmCdc45 localises to the chorion amplification foci in the follicle cells of the ovary. PMID- 11024169 TI - Minimising the secondary structure of DNA targets by incorporation of a modified deoxynucleoside: implications for nucleic acid analysis by hybridisation. AB - Some regions of nucleic acid targets are not accessible to heteroduplex formation with complementary oligonucleotide probes because they are involved in secondary structure through intramolecular Watson-Crick pairing. The secondary conformation of the target may be destabilised to assist its interaction with oligonucleotide probes. To achieve this, we modified a DNA target, which has self-complementary sequence able to form a hairpin loop, by replacing dC with N:4-ethyldeoxycytidine (d(4Et)C), which hybridises specifically with natural dG to give a G:(4Et)C base pair with reduced stability compared to the natural G:C base pair. Substitution by d(4Et)C greatly reduced formation of the target secondary structure. The lower level of secondary structure allowed hybridisation with complementary probes made with natural bases. We confirmed that hybridisation could be further enhanced by modifying the probes with intercalating groups which stabilise the duplex. PMID- 11024170 TI - PCR performance of the B-type DNA polymerase from the thermophilic euryarchaeon Thermococcus aggregans improved by mutations in the Y-GG/A motif. AB - The effect of mutations in the highly conserved Y-GG/A motif of B-type DNA polymerases was studied in the DNA polymerase from the hyperthermophilic euryarchaeon Thermococcus aggregans. This motif plays a critical role in the balance between the synthesis and degradation of the DNA chain. Five different mutations of the tyrosine at position 387 (Tyr387-->Phe, Tyr387-->Trp, Tyr387- >His, Tyr387-->Asn and Tyr387-->Ser) revealed that an aromatic ring system is crucial for the synthetic activity of the enzyme. Amino acids at this position lacking the ring system (Ser and Asn) led to a significant decrease in polymerase activity and to enhanced exonuclease activity, which resulted in improved enzyme fidelity. Exchange of tyrosine to phenylalanine, tryptophan or histidine led to phenotypes with wild-type-like fidelity but enhanced PCR performance that could be related to a higher velocity of polymerisation. With the help of a modelled structure of T.aggregans DNA polymerase, the biochemical data were interpreted proposing that the conformation of the flexible loop containing the Y-GG/A motif is an important factor for the equilibrium between DNA polymerisation and exonucleolysis. PMID- 11024172 TI - Characterization of the cold stress-induced cyanobacterial DEAD-box protein CrhC as an RNA helicase. AB - We have shown previously that CrhC is a unique member of the DEAD-box family of RNA helicases whose expression occurs specifically under conditions of cold stress. Here we show that recombinant His-tagged CrhC, purified from Escherichia coli, is an ATP-independent RNA binding protein possessing RNA-dependent ATPase activity which is stimulated most efficiently by rRNA and polysome preparations. RNA strand displacement assays indicate that CrhC possesses RNA unwinding activity that is adenosine nucleotide specific. Unwinding of partially duplexed RNA proceeds in the 5'-->3' but not the 3'-->5' direction using standard assay conditions. Immunoprecipitation and far-western analysis indicate that CrhC is a component of a multisubunit complex, interacting specifically with a 37 kDa polypeptide. We propose that CrhC unwinds cold-stabilized secondary structure in the 5'-UTR of RNA during cold stress. PMID- 11024171 TI - Activation of the BRLF1 promoter and lytic cycle of Epstein-Barr virus by histone acetylation. AB - Histone acetylation alters the chromatin structure and activates the genes that are repressed by histone deacetylation. This investigation demonstrates that treating P3HR1 cells with trichostatin A (TSA) activates the Epstein-Barr virus (EBV) lytic cycle, allowing the virus to synthesize three viral lytic proteins Rta, Zta and EA-D. Experimental results indicate that TSA and 12-O: tetradecanoylphorbol-13-acetate synergistically activate the transcription of BRLF1, an immediate-early gene of EBV. Chromatin immunoprecipitation assay reveals that histone H4 at the BRLF1 promoter is acetylated after P3HR1 cells are treated with TSA, suggesting that histone acetylation activates BRLF1 transcription. Furthermore, results in this study demonstrate that mutation of a YY1-binding site in the BRLF1 promoter activates BRLF1 transcription 1.6- and 2.3 fold in P3HR1 cells and C33A cells, respectively. Real time PCR analysis reveals that the mutation also increases the histone acetylation level of the nucleosomes at the BRLF1 promoter 1. 64- and 3.08-fold in P3HR1 and C33A cells, respectively. Results presented herein suggest that histone deacetylation plays an important role in maintaining the viral latency and histone acetylation at the BRLF1 promoter allows the virus to express Rta and to activate the viral lytic cycle. PMID- 11024173 TI - Transcription efficiency of human polymerase III genes in vitro does not depend on the RNP-forming autoantigen La. AB - Transcription of class III genes is conducted by multi-protein complexes consisting of polymerase III itself and several transcription factors. We established a reconstituted in vitro transcription system from which the autoantigen La was removed by immunodepletion. This system showed no RNP formation, but was still fully active in transcription. Supplementing such La free transcription reactions with recombinant La restored the formation of La complexes with the newly synthesised RNA, but did not lead to enhanced transcription efficiency. Furthermore, we developed a technique for the generation and isolation of transcription complexes, assembled from purified transcription factors and isolated by glycerol centrifugation. These complexes were fully competent to re-initiate RNA synthesis but they were not associated with La and their transcription rate could not be stimulated by addition of recombinant La. Therefore, we conclude that La does not act as a human polymerase III transcription factor. PMID- 11024174 TI - Expansion of the (CTG)(n) repeat in the 5'-UTR of a reporter gene impedes translation. AB - Effects of d(CAG)(n).d(CTG)(n) repeats on expression of a reporter gene in human cell culture were studied using transient transfection, RNase protection and coupled transcription/translation assays. Cloning these repeats into the reporter 3'-UTR did not affect gene functioning. In contrast, placing the repeats in the reporter 5'-UTR led to strong inhibition of expression. This inhibition depended on the repeat orientation, being prominent only when the (CTG)(n) tracts were in the sense strand for transcription. Further, the strength of inhibition increased exponentially with an increase in repeat length. Our data indicate that expanded (CTG)(n) repeats prevent efficient translation of the reporter mRNA both in vitro and in vivo. We suggest that formation of stable hairpins by (CUG)(n) runs of increasing length in the 5'-UTR of a mRNA progressively inhibits the scanning step of translation initiation. This points to a novel mechanism of regulating gene expression by expandable d(CTG)(n) repeats. PMID- 11024176 TI - Substrate binding in vitro and kinetics of RsrI [N6-adenine] DNA methyltransferase. AB - RSR:I [N:6-adenine] DNA methyltransferase (M.RSR:I), which recognizes GAATTC and is a member of a restriction-modification system in Rhodobacter sphaeroides, was purified to >95% homogeneity using a simplified procedure involving two ion exchange chromatographic steps. Electrophoretic gel retardation assays with purified M.RSR:I were performed on unmethylated, hemimethylated, dimethylated or non-specific target DNA duplexes (25 bp) in the presence of sinefungin, a potent inhibitory analog of AdoMet. M. RSR:I binding was affected by the methylation status of the DNA substrate and was enhanced by the presence of the cofactor analog. M. RSR:I bound DNA substrates in the presence of sinefungin with decreasing affinities: hemimethylated > unmethylated > dimethylated >> non specific DNA. Gel retardation studies with DNA substrates containing an abasic site substituted for the target adenine DNA provided evidence consistent with M.RSR:I extruding the target base from the duplex. Consistent with such base flipping, an approximately 1.7-fold fluorescence intensity increase was observed upon stoichiometric addition of M.RSR:I to hemimethylated DNA containing the fluorescent analog 2-aminopurine in place of the target adenine. Pre-steady-state kinetic and isotope- partitioning experiments revealed that the enzyme displays burst kinetics, confirmed the catalytic competence of the M.RSR:I-AdoMet complex and eliminated the possibility of an ordered mechanism where DNA is required to bind first. The equilibrium dissociation constants for AdoMet, AdoHcy and sinefungin were determined using an intrinsic tryptophan fluorescence-quenching assay. PMID- 11024177 TI - DNA binding properties in vivo and target recognition domain sequence alignment analyses of wild-type and mutant RsrI [N6-adenine] DNA methyltransferases. AB - A genetic selection method, the P22 challenge-phage assay, was used to characterize DNA binding in vivo by the prokaryotic beta class [N:6-adenine] DNA methyltransferase M.RSR:I. M.RSR:I mutants with altered binding affinities in vivo were isolated. Unlike the wild-type enzyme, a catalytically compromised mutant, M.RSR:I (L72P), demonstrated site-specific DNA binding in vivo. The L72P mutation is located near the highly conserved catalytic motif IV, DPPY (residues 65-68). A double mutant, M.RSR:I (L72P/D173A), showed less binding in vivo than did M.RSR:I (L72P). Thus, introduction of the D173A mutation deleteriously affected DNA binding. D173 is located in the putative target recognition domain (TRD) of the enzyme. Sequence alignment analyses of several beta class MTases revealed a TRD sequence element that contains the D173 residue. Phylogenetic analysis suggested that divergence in the amino acid sequences of these methyltransferases correlated with differences in their DNA target recognition sequences. Furthermore, MTases of other classes (alpha and gamma) having the same DNA recognition sequence as the beta class MTases share related regions of amino acid sequences in their TRDs. PMID- 11024175 TI - Structure of RsrI methyltransferase, a member of the N6-adenine beta class of DNA methyltransferases. AB - DNA methylation is important in cellular, developmental and disease processes, as well as in bacterial restriction-modification systems. Methylation of DNA at the amino groups of cytosine and adenine is a common mode of protection against restriction endonucleases afforded by the bacterial methyltransferases. The first structure of an N:6-adenine methyltransferase belonging to the beta class of bacterial methyltransferases is described here. The structure of M. RSR:I from Rhodobacter sphaeroides, which methylates the second adenine of the GAATTC sequence, was determined to 1.75 A resolution using X-ray crystallography. Like other methyltransferases, the enzyme contains the methylase fold and has well defined substrate binding pockets. The catalytic core most closely resembles the PVU:II methyltransferase, a cytosine amino methyltransferase of the same beta group. The larger nucleotide binding pocket observed in M. RSR:I is expected because it methylates adenine. However, the most striking difference between the RSR:I methyltransferase and the other bacterial enzymes is the structure of the putative DNA target recognition domain, which is formed in part by two helices on an extended arm of the protein on the face of the enzyme opposite the active site. This observation suggests that a dramatic conformational change or oligomerization may take place during DNA binding and methylation. PMID- 11024178 TI - Soggy, a spermatocyte-specific gene, lies 3.8 kb upstream of and antipodal to TEAD-2, a transcription factor expressed at the beginning of mouse development. AB - Investigation of the regulatory region of mTEAD-2, a gene expressed at the beginning of mouse pre-implantation development, led to the surprising discovery of another gene only 3.8 kb upstream of mTEAD-2. Here we show that this new gene is a single copy, testis-specific gene called SOGGY: (mSgy) that produces a single, dominant mRNA approximately 1.3 kb in length. It is transcribed in the direction opposite to mTEAD-2, thus placing the regulatory elements of these two genes in close proximity. mSgy contains three methionine codons that could potentially act as translation start sites, but most mSGY protein synthesis in vitro was initiated from the first Met codon to produce a full-length protein, suggesting that mSGY normally consists of 230 amino acids (26.7 kDa). Transcription began at a cluster of nucleotides approximately 150 bp upstream of the first Met codon using a TATA-less promoter contained within the first 0.9 kb upstream. The activity of this promoter was repressed by upstream sequences between -0.9 and -2.5 kb in cells that did not express mSgy, but this repression was relieved in cells that did express mSgy. mSgy mRNA was detected in embryos only after day 15 and in adult tissues only in the developing spermatocytes of seminiferous tubules, suggesting that mSgy is a spermatocyte-specific gene. Since mTEAD-2 and mSgy were not expressed in the same cells, the mSgy/mTEAD-2 locus provides a unique paradigm for differential regulation of gene expression during mammalian development. PMID- 11024179 TI - Cell cycle-independent removal of UV-induced pyrimidine dimers from the promoter and the transcription initiation domain of the human CDC2 gene. AB - To assess whether removal of UV-induced cyclobutane pyrimidine dimers (CPDs) occurs with equal efficiency at different stages of the cell cycle in a cell cycle-regulated gene, we have analyzed repair of CPDs, following a single dose of UV, in normal human fibroblasts that were synchronized in either G(0) or S phase. Based on a single nucleotide resolution analysis, we established a detailed map of DNA repair rates along the promoter region and the transcription initiation area of the human CDC2 gene. The promoter of this gene is covered by an array of sequence-specific transcription factors located between nt -280 and -9 relative to the major transcription start site. In both quiescent and S phase-synchronized fibroblasts the majority of these sequences were poorly repaired even after 24 h, probably as a result of the constitutive binding of transcription factors throughout the cell cycle. A domain of fast repair was found at sequences surrounding the transcription initiation site and continuing downstream for approximately 80 nt. CPD removal from this domain was preferential in both quiescent and proliferating fibroblasts, despite lower levels of global genome repair and a lack of CDC2 transcription in quiescent cells. We suggest that sequences involved in transcription initiation may be book-marked for efficient repair throughout the cell cycle, even when the gene is temporarily not expressed. PMID- 11024180 TI - Elongation of repetitive DNA by DNA polymerase from a hyperthermophilic bacterium Thermus thermophilus. AB - Short repetitive DNA sequences are believed to be one of the primordial genetic elements that served as a source of complex large DNA found in the genome of modern organisms. However, the mechanism of its expansion (increase in repeat number) during the course of evolution is unclear. We demonstrate that the DNA polymerase of the hyperthermophilic bacterium Thermus thermophilus can elongate oligoDNA with several tandem repeats to very long DNA in vitro. For instance, 48mer repetitive oligoDNA (TACATGTA)(6), which has 25% GC content and a palindromic sequence, can be elongated up to approximately 10 000 bases by DNA polymerase at 74 degrees C without template DNA. OligoDNA having a different GC content or a quasi-palindromic sequence can also be elongated, but less efficiently. A spectroscopic thermal melting experiment with the oligoDNA showed that its hairpin-coil transition temperature was very close to the elongation reaction temperature (74 degrees C), but was much higher than the temperature at which duplex oligoDNA can exist stably. Taken together, we conclude that repetitive oligoDNA with a palindromic or quasi-palindromic sequence is elongated extensively by a hyperthermophilic DNA polymerase through hairpin-coil transitions. We propose that such an elongation mechanism might have been a driving force to expand primordial short DNA. PMID- 11024181 TI - p53 C-terminal interaction with DNA ends and gaps has opposing effect on specific DNA binding by the core. AB - In addition to binding DNA in a sequence-specific manner, the p53 tumour suppressor protein can interact with damaged DNA. In order to understand which structural features in DNA the C-teminal domain recognises we have studied the interaction of p53 protein with different types of DNA oligonucleotides imitating damaged DNA. Here we show that one unpaired nucleotide within double-stranded (ds)DNA is sufficient for recognition by the p53 C-terminus, either as a protruding end or as an internal gap in dsDNA. C-terminal interaction with DNA ends facilitated core domain binding to DNA, whereas interaction with gaps prevented core domain-DNA complexing, implying that p53 might adopt distinct conformations upon binding to different DNA lesions. These observations suggest that both single-strand and double-strand breaks can serve as a target for p53 C terminal recognition in vivo and indicate that p53 might recruit different repair factors to the sites of damaged DNA depending on the type of lesion. PMID- 11024183 TI - A heuristic graph comparison algorithm and its application to detect functionally related enzyme clusters. AB - The availability of computerized knowledge on biochemical pathways in the KEGG database opens new opportunities for developing computational methods to characterize and understand higher level functions of complete genomes. Our approach is based on the concept of graphs; for example, the genome is a graph with genes as nodes and the pathway is another graph with gene products as nodes. We have developed a simple method for graph comparison to identify local similarities, termed correlated clusters, between two graphs, which allows gaps and mismatches of nodes and edges and is especially suitable for detecting biological features. The method was applied to a comparison of the complete genomes of 10 microorganisms and the KEGG metabolic pathways, which revealed, not surprisingly, a tendency for formation of correlated clusters called FRECs (functionally related enzyme clusters). However, this tendency varied considerably depending on the organism. The relative number of enzymes in FRECs was close to 50% for Bacillus subtilis and Escherichia coli, but was <10% for SYNECHOCYSTIS: and Saccharomyces cerevisiae. The FRECs collection is reorganized into a collection of ortholog group tables in KEGG, which represents conserved pathway motifs with the information about gene clusters in all the completely sequenced genomes. PMID- 11024182 TI - Hammerhead ribozymes selectively suppress mutant type I collagen mRNA in osteogenesis imperfecta fibroblasts. AB - Ribozymes are a promising agent for the gene therapy of dominant negative genetic disorders by allele-specific mRNA suppression. To test allele-specific mRNA suppression in cells, we used fibroblasts from a patient with osteogenesis imperfecta (OI). These cells contain a mutation in one alpha1(I) collagen allele which both causes the skeletal disorder and generates a novel ribozyme cleavage site. In a preliminary in vitro assay, ribozymes cleaved mutant RNA substrate whereas normal substrate was left intact. For the studies in cell culture we generated cell lines stably expressing active (AR) and inactive (IR) ribozymes targeted to mutant alpha1(I) collagen mRNA. Quantitative competitive RT-PCR analyses of type I collagen mRNA, normalized to beta-actin expression levels, revealed that the level of mutant alpha1(I) collagen mRNA was significantly decreased by approximately 50% in cells expressing AR. Normal alpha1(I) collagen mRNA showed no significant reduction when AR or IR was expressed from the pHbetaAPr-1-neo vector and a small (10-20%) but significant reduction when either ribozyme was expressed from the pCI.neo vector. In clonal lines derived from cells expressing AR the level of ribozyme expression correlated with the extent of reduction in the mutant:normal alpha1(I) mRNA ratio, ranging from 0.33 to 0.96. Stable expression of active ribozyme did not affect cell viability, as assessed by growth rates. Ribozyme cleavage of mutant mRNA results in a reduction in mutant type I collagen protein, as demonstrated by SDS-urea-PAGE. This is the first report of ribozymes causing specific suppression of an endogenous mutant mRNA in cells derived from a patient with a dominant negative genetic disorder. PMID- 11024184 TI - Automatic detection of conserved gene clusters in multiple genomes by graph comparison and P-quasi grouping. AB - We previously reported two graph algorithms for analysis of genomic information: a graph comparison algorithm to detect locally similar regions called correlated clusters and an algorithm to find a graph feature called P-quasi complete linkage. Based on these algorithms we have developed an automatic procedure to detect conserved gene clusters and align orthologous gene orders in multiple genomes. In the first step, the graph comparison is applied to pairwise genome comparisons, where the genome is considered as a one-dimensionally connected graph with genes as its nodes, and correlated clusters of genes that share sequence similarities are identified. In the next step, the P-quasi complete linkage analysis is applied to grouping of related clusters and conserved gene clusters in multiple genomes are identified. In the last step, orthologous relations of genes are established among each conserved cluster. We analyzed 17 completely sequenced microbial genomes and obtained 2313 clusters when the completeness parameter P: was 40%. About one quarter contained at least two genes that appeared in the metabolic and regulatory pathways in the KEGG database. This collection of conserved gene clusters is used to refine and augment ortholog group tables in KEGG and also to define ortholog identifiers as an extension of EC numbers. PMID- 11024185 TI - Hammerhead-mediated processing of satellite pDo500 family transcripts from Dolichopoda cave crickets. AB - This work reports the discovery and functional characterization of catalytically active hammerhead motifs within satellite DNA of the pDo500 family from several DOLICHOPODA: cave cricket species. We show that in vitro transcribed RNA of some members of this satellite DNA family do self-cleave in vitro. This self-cleavage activity is correlated with the efficient in vivo processing of long primary transcripts into monomer-sized RNA. The high sequence conservation of the satellite pDo500 DNA family among genetically isolated DOLICHOPODA: schiavazzii populations, as well as other DOLICHOPODA: species, along with the fact that satellite members are actively transcribed in vivo suggests that the hammerhead encoding satellite transcripts are under selective pressure, perhaps because they fulfil an important physiological role or function. Remarkably, this is the third example of hammerhead ribozyme structures associated with transcribed repetitive DNA sequences from animals. The possibility that such an association may not be purely coincidental is discussed. PMID- 11024186 TI - DNA binding by single HMG box model proteins. AB - The HMG1/2 family is a large group of proteins that share a conserved sequence of approximately 80 amino acids rich in basic, aromatic and proline side chains, referred to as an HMG box. Previous studies show that HMG boxes can bind to DNA in a structure-specific manner. To define the basis for DNA recognition by HMG boxes, we characterize the interaction of two model HMG boxes, one a structure specific box, rHMGb from the rat HMG1 protein, the other a sequence-specific box, Rox1 from yeast, with oligodeoxynucleotide substrates. Both proteins interact with single-stranded oligonucleotides in this study to form 1:1 complexes. The stoichiometry of binding of rHMGb to duplex or branched DNAs differs: for a 16mer duplex we find a weak 2:1 complex, while a 4:1 protein:DNA complex is detected with a four-way DNA junction of 16mers in the presence of Mg(2+). In the case of the sequence-specific Rox1 protein we find tight 1:1 and 2:1 complexes with its cognate duplex sequence and again a 4:1 complex with four-way branched DNA. If the DNA branching is reduced to three arms, both proteins form 3:1 complexes. We believe that these multimeric complexes are relevant for HMG1/2 proteins in vivo, since Mg(2+) is present in the nucleus and these proteins are expressed at a very high level. PMID- 11024188 TI - Rapid determination of short DNA sequences by the use of MALDI-MS. AB - We have developed a protocol for rapid sequencing of short DNA stretches (15-20 nt) using MALDI-TOF-MS. The protocol is based on the Sanger concept with the modification that double-stranded template DNA is used and all four sequencing reactions are performed in one reaction vial. The sequencing products are separated and detected by MALDI-TOF-MS and the sequence is determined by comparing measured molecular mass differences to expected values. The protocol is optimized for low costs and broad applicability. One reaction typically includes 300 fmol template, 10 pmol primer and 200 pmol each nucleotide monomer. Neither the primer nor any of the nucleotide monomers are labeled. Solid phase purification, concentration and mass spectrometric sample preparation of the sequencing products are accomplished in a few minutes and parallel processing of 96 samples is possible. The mass spectrometric analyses and subsequent sequence read-out require only a few seconds per template. PMID- 11024187 TI - Amplification of IgG VH and VL (Fab) from single human plasma cells and B cells. AB - Amplification of human immunoglobulin has many potential applications such as analysis of clonality, isolation of immunogenic antigens and antigen-specific immunotherapy. Here we describe a method for amplification of human immunoglobulin heavy and light chains from single B lymphocytes or plasma cells. Cells are isolated by FACS, and Ig is amplified by semi-nested RT-PCR. The method is versatile, sensitive and reliable: it provides appropriately paired heavy and light chains, requiring as little as 2 days to produce amplified Fab DNA from human tissues. PMID- 11024189 TI - Solid phase DNA amplification: characterisation of primer attachment and amplification mechanisms. AB - Different chemical methods used to attach oligonucleotides by their 5'-end on a glass surface were tested in the framework of solid phase PCR where surface-bound instead of freely-diffusing primers are used to amplify DNA. Each method was first evaluated for its capacity to provide a high surface coverage of oligonucleotides essentially attached via a 5'-specific linkage that satisfyingly withstands PCR conditions and leaves the 3'-ends available for DNA polymerase activity. The best results were obtained with 5'-thiol-modified oligonucleotides attached to amino-silanised glass slides using a heterobifunctional cross-linker reagent. It was then demonstrated that the primers bound to the glass surface using the optimal chemistry can be involved in attaching and amplifying DNA molecules present in the reaction mix in the absence of freely-diffusing primers. Two distinct amplification processes called interfacial and surface amplification have been observed and characterised. The newly synthesised DNA can be detected and quantified by radioactive and fluorescent hybridisation assays. These new surface amplification processes are seen as an interesting approach for attachment of DNA molecules by their 5'-end on a solid support and can be used as an alternative route for producing DNA chips for genomic studies. PMID- 11024190 TI - High efficiency family shuffling based on multi-step PCR and in vivo DNA recombination in yeast: statistical and functional analysis of a combinatorial library between human cytochrome P450 1A1 and 1A2. AB - The design of a family shuffling strategy (CLERY: Combinatorial Libraries Enhanced by Recombination in Yeast) associating PCR-based and in vivo recombination and expression in yeast is described. This strategy was tested using human cytochrome P450 CYP1A1 and CYP1A2 as templates, which share 74% nucleotide sequence identity. Construction of highly shuffled libraries of mosaic structures and reduction of parental gene contamination were two major goals. Library characterization involved multiprobe hybridization on DNA macro-arrays. The statistical analysis of randomly selected clones revealed a high proportion of chimeric genes (86%) and a homogeneous representation of the parental contribution among the sequences (55.8 +/- 2.5% for parental sequence 1A2). A microtiter plate screening system was designed to achieve colorimetric detection of polycyclic hydrocarbon hydroxylation by transformed yeast cells. Full sequences of five randomly picked and five functionally selected clones were analyzed. Results confirmed the shuffling efficiency and allowed calculation of the average length of sequence exchange and mutation rates. The efficient and statistically representative generation of mosaic structures by this type of family shuffling in a yeast expression system constitutes a novel and promising tool for structure-function studies and tuning enzymatic activities of multicomponent eucaryote complexes involving non-soluble enzymes. PMID- 11024191 TI - Mutation analysis of the entire mitochondrial genome using denaturing high performance liquid chromatography. AB - In patients with mitochondrial disease a continuously increasing number of mitochondrial DNA (mtDNA) mutations and polymorphisms have been identified. Most pathogenic mtDNA mutations are heteroplasmic, resulting in heteroduplexes after PCR amplification of mtDNA. To detect these heteroduplexes, we used the technique of denaturing high performance liquid chromatography (DHPLC). The complete mitochondrial genome was amplified in 13 fragments of 1-2 kb, digested in fragments of 90-600 bp and resolved at their optimal melting temperature. The sensitivity of the DHPLC system was high with a lowest detection of 0.5% for the A8344G mutation. The muscle mtDNA from six patients with mitochondrial disease was screened and three mutations were identified. The first patient with a limb girdle-type myopathy carried an A3302G substitution in the tRNA(Leu(UUR)) gene (70% heteroplasmy), the second patient with mitochondrial myopathy and cardiomyopathy carried a T3271C mutation in the tRNA(Leu(UUR)) gene (80% heteroplasmy) and the third patient with Leigh syndrome carried a T9176C mutation in the ATPase6 gene (93% heteroplasmy). We conclude that DHPLC analysis is a sensitive and specific method to detect heteroplasmic mtDNA mutations. The entire automatic procedure can be completed within 2 days and can also be applied to exclude mtDNA involvement, providing a basis for subsequent investigation of nuclear genes. PMID- 11024192 TI - Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization. AB - Suppression subtractive hybridization (SSH) is one of the most powerful and popular methods for isolating differentially expressed transcripts. However, SSH generated libraries typically contain some background clones representing non differentially expressed transcripts. To overcome this problem we developed a simple procedure that substantially decreases the number of background clones. This method is based on the following difference between target and background cDNAs: each kind of background molecule has only one orientation with respect to the two different flanking adapter sequences used in SSH, while truly differentially expressed target cDNA fragments are represented by both sequence orientations. The described method selects the molecules that arose due to hybridization of such mirror-orientated molecules. The efficiency of this method was demonstrated in both model and real experimental subtractions. PMID- 11024193 TI - DNA probes on chip surfaces studied by scanning force microscopy using specific binding of colloidal gold. AB - Single-stranded DNA was covalently bound on chip surfaces using two different silanization procedures. The resulting surfaces were characterized by fluorescence and scanning force microscopy using sequence-complementary DNA molecules with labels. Colloidal gold (30 nm) was used as the topographic label. Scanning force microscopy revealed the individual labels on the surface and their distribution. Steps of silane layers or DNA-modified surfaces prepared using an elastomeric mask provided internal controls for comparison of modified with unmodified surfaces. PMID- 11024194 TI - Evaluation of environmental bacterial contamination and procedures to control cross infection in a sample of Italian dental surgeries. AB - OBJECTIVES: To perform a pilot study on bacterial contamination in some dental surgeries (n=51) in a local health unit in Brescia (Lombardy Region, Italy) and to evaluate the procedures to control cross infection used by the personnel to reduce the risk of infection in dental practice. METHODS: A survey was carried out by interviewing 133 dental personnel with a questionnaire on the procedures used to control infection. The autoclaves, chemical baths (chemiclaves), and ovens present in the surgeries were tested for sterilisation efficiency with a spore test, and already packed and sterilised instruments were randomly sampled and tested for sterility. Microbial contamination of air, surface, and dental unit water samples were also studied. RESULTS: The dental personnel did not generally follow the principal procedures for infection control: 30% of personnel were not vaccinated against hepatitis B virus, infected instruments were often not decontaminated, periodic checks of autoclave efficiency were lacking, and the knowledge of disinfection mechanisms and procedures was incomplete. High bacteriological contamination of water at dental surgeries was often found and total bacteriological counts in air samples were high. Surface studies showed widespread bacterial contamination. CONCLUSIONS: On the basis of these results, an educational programme for the prevention of infective hazards has been prepared and carried out. The results of this pilot study will be used for planning a national survey. PMID- 11024195 TI - In vitro toxicology of respirable Montserrat volcanic ash. AB - OBJECTIVES: In July 1995 the Soufriere Hills volcano on the island of Montserrat began to erupt. Preliminary reports showed that the ash contained a substantial respirable component and a large percentage of the toxic silica polymorph, cristobalite. In this study the cytotoxicity of three respirable Montserrat volcanic ash (MVA) samples was investigated: M1 from a single explosive event, M2 accumulated ash predominantly derived from pyroclastic flows, and M3 from a single pyroclastic flow. These were compared with the relatively inert dust TiO(2) and the known toxic quartz dust, DQ12. METHODS: Surface area of the particles was measured with the Brunauer, Emmet, and Teller (BET) adsorption method and cristobalite content of MVA was determined by x ray diffraction (XRD). After exposure to particles, the metabolic competence of the epithelial cell line A549 was assessed to determine cytotoxic effects. The ability of the particles to induce sheep blood erythrocyte haemolysis was used to assess surface reactivity. RESULTS: Treatment with either MVA, quartz, or titanium dioxide decreased A549 epithelial cell metabolic competence as measured by ability to reduce 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). On addition of mannitol, the cytotoxic effect was significantly less with M1, quartz, and TiO(2). All MVA samples induced a dose dependent increase in haemolysis, which, although less than the haemolysis induced by quartz, was significantly greater than that induced by TiO(2). Addition of mannitol and superoxide dismutase (SOD) significantly reduced the haemolytic activity only of M1, but not M2 or M3, the samples derived from predominantly pyroclastic flow events. CONCLUSIONS: Neither the cristobalite content nor the surface area of the MVA samples correlated with observed in vitro reactivity. A role for reactive oxygen species could only be shown in the cytotoxicity of M1, which was the only sample derived from a purely explosive event. These results suggest that in general the bioreactivity of MVA samples in vitro is low compared with pure quartz, but that the bioreactivity and mechanisms of biological interaction may vary according to the ash source. PMID- 11024196 TI - Dermal in vitro penetration of methiocarb, paclobutrazol, and pirimicarb. AB - OBJECTIVES: The dominant route of occupational exposure to pesticides in horticulture is dermal. However, preventive measures are seldom used when handling plant cultures recently treated with pesticides, thus causing significant dermal exposure and potential absorption. Assessment of exposure often depends on biological monitoring of blood or urine samples. The skin often acts as a temporary reservoir for chemicals before absorption. Failure to consider the lag time between dermal exposure and appearance of pesticide or metabolites in the general circulation may lead to false conclusions about assessment of exposure. METHODS: In an experimental model in which in vitro static diffusion cells were mounted with human skin, dermal penetration of three extensively used pesticides (methiocarb, paclobutrazol, pirimicarb) was evaluated. RESULTS: Pirimicarb and paclobutrazol had comparable rates of dermal penetration and lag times of around 18 hours. Methiocarb had a considerably shorter lag time. Dermal penetration continued for extended periods after exposure had ended. CONCLUSIONS: With lag times sometimes considerably longer than a normal working day, biological monitoring at the end of exposure may seriously underestimate the actual exposure. There may be implications for regulatory guidelines, which often require only 24 hour observation periods. PMID- 11024197 TI - Personal air sampling and biological monitoring of occupational exposure to the soil fumigant cis-1,3-dichloropropene. AB - OBJECTIVES: To assess exposure of commercial application workers to the nematocide cis-1,3-dichloropropene (cis-DCP). METHODS: The study was conducted during the annual application season, August to 15 November, in the starch potato growing region in The Netherlands. 14 Application workers collected end of shift urine samples on each fumigation day (n=119). The mercapturic acid metabolite N acetyl-S-(cis-3-chloro-2-propenyl)-L-cysteine (cis-DCP-MA) in urine was used for biological monitoring of the cis-DCP uptake. Inhalatory exposure was assessed by personal air sampling during a representative sample (n=37) of the fumigation days. Extensive information was collected on factors of possible relevance to the exposure and the application workers were observed for compliance with the statutory directions for use. The inhalatory exposure during all fumigation days was estimated from the relation between the personal air sampling data and the biological monitoring data. Exposure levels were correlated with the general work practice. The fumigation equipment and procedures were in accordance with the statutory directions of use, with the exception of the antidrip systems. Two antidrip systems were used: antidrip nozzles or a compressed air system. RESULTS: The geometric mean exposure of the application workers was 2.7 mg/m(3) (8 hour time weighted average); range 0.1-9.5 mg/m(3). On 25 days (21%) the exposure exceeded the Dutch occupational exposure limit (OEL) of 5 mg/m(3). This could mainly be explained by prolonged working days of more than 8 hours. The general work practice of the application workers was rated by the observers as good or poor. No difference in exposure to cis-DCP was found in the use of none, one, or two antidrip systems. Malfunctioning of the antidrip systems and lack of experience with the compressed air system were identified as possible causes for the lack of effectiveness of these antidrip systems. The use of personal protection was not always in accordance with the statutory directions of use. Dermal exposure to liquid cis-DCP was found four times during repair and maintenance, but the biological monitoring data did not suggest a significant increase in cis-DCP uptake. CONCLUSIONS: The application of cis-DCP in the potato growing industry can be performed at exposure concentrations below the Dutch OEL of 5 mg/m(3) if the working days are limited to 8 hours. An injector equipped with either kind of antidrip system which is in good working order, as well as the consistent use of personal protection in accordance with the statutory directions of use, may ensure exposure concentrations below the Dutch OEL. PMID- 11024199 TI - Survey of symptoms, respiratory function, and immunology and their relation to glutaraldehyde and other occupational exposures among endoscopy nursing staff. AB - OBJECTIVES: To find the nature and incidence of symptoms experienced by a large sample of hospital endoscopy nurses. To find whether nurses in endoscopy units develop asthma under current working conditions in endoscopy units. To obtain analytically reliable data on exposure concentrations of glutaraldehyde (GA) vapour in endoscopy units, and to relate them to individual hygiene and work practices. To characterise any exposure-response relations between airborne GA and the occurrence of work related symptoms (WRSs). Due to the growing concern about the perceived increase in WRSs among workers regularly exposed to biocides, all of whom work within a complex multiexposure environment, a cross sectional study was designed. METHODS: Current endoscopy nurses (n=348) from 59 endoscopy units within the United Kingdom and ex-employees (who had left their job for health reasons (n=18) were surveyed. Symptom questionnaires, end of session spirometry, peak flow diaries, skin prick tests (SPTs) to latex and common aeroallergens, and measurements of total immunoglobulin E (IgE) and IgE specific to GA and latex were performed. Exposure measurements included personal airborne biocide sampling for peak (during biocide changeover) and background (endoscopy room, excluding biocide changeover) concentrations. RESULTS: All 18 ex-employees and 91.4% of the current nurses were primarily exposed to GA, the rest were exposed to a succinaldehyde-formaldehyde (SF) composite. Work related contact dermatitis was reported by 44% of current workers exposed to GA, 56.7% of those exposed to SF composite, and 44.4% of ex-employees. The prevalence of WRSs of the eyes, nose, and lower respiratory tract in current workers exposed to GA was 13.5%, 19.8%, and 8.5% respectively and 50%, 61.1%, and 66.6% in the ex employees. The mean percentage predicted forced expired volume in 1 second (ppFEV(1)) for ex-employees (93.82, 95% confidence interval (95% CI) 88.53 to 99.11) was significantly lower (p<0.01) than that of current workers exposed to GA (104.08, 95% CI 102.35 to 105.73). Occupational peak flow diaries completed by current workers with WRSs of the lower respiratory tract showed no evidence of bronchial asthma (<15% variation). Six per cent of the population had positive latex SPTs. Positive indications of one GA specific IgE and 4.1% latex specific IgE occurred. There was no conformity between the latex specific IgE and positive SPTs. Positive SPTs to latex were associated with WRSs of dermatitis and ocular WRSs, but no other WRSs. Exposures were above the current maximum exposure limit (MEL) of 0.2 mg/m(3) (0.05 ppm) in eight of the units investigated. A significant relation existed between peak GA concentrations and work related chronic bronchitis and nasal symptoms (after adjustment for types of local ventilation) but not to other WRSs. Peak GA concentrations were significantly higher in units that used both negative pressure room and decontaminating unit ventilation. CONCLUSION: This study documents a significant level of symptoms reported in the absence of objective evidence of the physiological changes associated with asthma. Ex-employees and current workers with WRSs warrant further study to elucidate the cause and mechanisms for their symptoms. Ventilation systems used for the extraction of aldehydes from the work area may be less effective than expected and due to poor design may even contribute to high peak exposures. PMID- 11024198 TI - Occupational exposure to cis-1,3-dichloropropene: biological effect monitoring of kidney and liver function. AB - OBJECTIVES: To investigate the possible effects of occupational exposure to the nematocide cis-1,3-dichloropropene (cis-DCP) on function of the kidney and liver in the starch potato growing region in The Netherlands. METHODS: The study involved 13 commercial application workers exposed to cis-DCP for 117 days, and 22 matched control workers. The inhalatory exposure of the application workers was estimated from biological monitoring data. All workers collected urine and serum samples before, during, and after the fumigation season for monitoring of variables for kidney and liver function. Renal effect variables were alanine aminopeptidase (AAP), N-acetyl-beta-D-glucosaminidase (NAG), retinol binding protein (RBP), and albumin (ALB) in urine, and beta(2)-microglobulin (beta(2)M-S) and creatinine in serum (Creat-S). Liver variables were alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), gamma-glutamyltranspeptidase (GGT), alkaline phosphatase (ALP), and total bilirubin (TBIL) in serum and the urinary ratio of 6-beta-hydroxycortisol to free cortisol (betaOHC/COR). RESULTS: The geometric mean exposure of the application workers was 2.7 mg/m(3) (8 hour time weighted average (8 hour TWA)); range 0.1-9.5 mg/m(3). No differences were found between the values of the renal effect variables or the liver variables of the exposed group and the control group, except a lower urinary ratio of betaOHC/COR in the exposed group. This was not considered to be related to the exposure to cis-DCP. No dose-effect relations were found between the exposure indices and the effect variables. CONCLUSIONS: The present study does not provide evidence that occupational exposure to cis-DCP in the starch potato growing region causes adverse effects on the kidney or liver at 8 hour TWA exposure concentrations below 9.5 mg/m(3) (2 ppm). PMID- 11024200 TI - Occupation and small bowel adenocarcinoma: a European case-control study. AB - OBJECTIVES: Because of the rarity of small bowel adenocarcinoma (SBA), little is known about the aetiology of this disease. This study aimed to identify occupational clustering of cases SBA as a systematic approach to new hypotheses on the aetiology of this disease. METHODS: A European multicentre case-control study was conducted in 1995-7, inclusive. Incident cases aged 35-69 years with SBA (n=168) were recruited before acceptance by a pathologist. Altogether 107 cases and 3915 controls were accepted, of which 79 cases, 579 colon cancer controls, and 2070 population controls were interviewed. RESULTS: The strongest industrial risk factors for SBA taking account of 10 years' exposure lag were dry cleaning, manufacture of workwear, mixed farming (women), and manufacture of motor vehicles (men). A significantly increased risk of SBA (odds ratio (OR) and 95% confidence interval (95% CI)) was found among men employed as building caretakers, OR 6.7 (1.7 to 26.0) and women employed as housekeepers, OR 2.2 (1.1 to 4.9); general farm labourers, OR 4.7 (1.8 to 12.2); dockers, OR 2.9 (1.0 to 8.2); dry cleaners or launderers, OR 4.1 (1.2 to 13.6); and textile workers (sewers or embroiderers), OR 2.6 (1.0 to 6.8). For the last four groups, together with welders OR 2.7 (1.1 to 6.6) (men) an exposure-response pattern was found when calculating the ORs for jobs held 1-5 years and >5 years, with never having held the job as reference. The ORs (95% CIs) for 1-5 years and >5 years were 4.3 (0.4 to 44.0) and 3.5 (0.9 to 13.7), 3.0 (0.3 to 26.2) and 4.3 (0.9 to 21.2), 4.6 (0.4 to 48.1) and 11.0 (2.0 to 60.4), 1.3 (0.2 to 11.0) and 5.8 (2.0 to 17.2), and 2.8 (0.3 to 23.8) and 4.6 (1.3 to 16.6), respectively, for each of these occupations. Among welders, people performing semiautomatic arc welding (MIG/MAG) were identified as a high risk group (OR 5.0 (1.3 to 19.6)). CONCLUSIONS: This explorative study suggests an increased occurrence of SBA in certain occupations, which needs further evaluation. PMID- 11024201 TI - Laryngeal and hypopharyngeal cancers and occupational exposure to formaldehyde and various dusts: a case-control study in France. AB - OBJECTIVES: A case-control study was conducted in France to assess possible associations between occupational exposures and squamous cell carcinomas of the larynx and hypopharynx. METHODS: The study was restricted to men, and included 201 hypopharyngeal cancers, 296 laryngeal cancers, and 296 controls (patients with other tumour sites). Detailed information on smoking, alcohol consumption, and lifetime occupational history was collected. Occupational exposure to seven substances (formaldehyde, leather dust, wood dust, flour dust, coal dust, silica dust, and textile dust) was assessed with a job exposure matrix. Exposure variables used in the analysis were probability, duration, and cumulative level of exposure. Odds ratios (ORs) with their 95% confidence intervals (95% CIs) were estimated by unconditional logistic regression, and were adjusted for major confounding factors (age, smoking, alcohol, and when relevant other occupational exposures). RESULTS: Hypopharyngeal cancer was found to be associated with exposure to coal dust (OR 2.31, 95% CI 1.21 to 4.40), with a significant rise in risk with probability (p<0.005 for trend) and level (p<0.007 for trend) of exposure. Exposure to coal dust was also associated with an increased risk of laryngeal cancer (OR 1.67, 95% CI 0.92 to 3.02), but no dose-response pattern was found. A significant relation, limited to hypopharyngeal cancer, was found with the probability of exposure to formaldehyde (p<0.005 for trend), with a fourfold risk for the highest category (OR 3.78, 95% CI 1.50 to 9.49). When subjects exposed to formaldehyde with a low probability were excluded, the risk also increased with duration (p<0.04) and cumulative level of exposure (p<0.14). No significant association was found for any other substance. CONCLUSION: These results indicate that exposure to formaldehyde and coal dust may increase the risk of hypopharyngeal cancer. PMID- 11024202 TI - Historical cohort study of 10 109 men in the North American vinyl chloride industry, 1942-72: update of cancer mortality to 31 December 1995. AB - OBJECTIVES: To update and assess mortality from neoplasms to 31 December 1995 among 10 109 men employed in a job exposed to vinyl chloride for at least 1 year between 1942 and 1972 at any of 37 North American factories. Previous analyses indicated associations between employment in vinyl production and increased mortality risk from cancers of the liver and biliary tract, due to increased mortality from angiosarcoma of the liver, and brain cancer. METHODS: Standardised mortality ratio (SMR) analyses, overall and stratified by several work related variables, were conducted with United States and state reference rates. Cox's proportional hazards models and stratified log rank tests were used to further assess occupational factors. RESULTS: 895 of 3191 deaths (28%) were from malignant neoplasms, 505 since the previous update to the end of 1982. Mortality from all causes showed a deficit (SMR 83, 95% confidence interval (95% CI) 80 to 86), whereas mortality from all cancers combined was similar to state referent rates. Mortality from cancers of the liver and biliary tract was clearly increased (SMR 359, 95% CI 284 to 446). Modest excesses of brain cancer (SMR 142, 95% CI 100 to 197) and cancer of connective and soft tissue (SMR 270, 95% CI 139 to 472) were found. Stratified SMR and Cox's proportional hazard analyses supported associations with age at first exposure, duration of exposure, and year of first exposure for cancers of the liver and soft tissues, but not the brain. CONCLUSIONS: Excess mortality risk from cancer of the liver and biliary tract, largely due to angiosarcoma, continues. Risk of mortality from brain cancer has attenuated, but its relation with exposure to vinyl chloride remains unclear. A potentially work related excess of deaths from cancer of connective and soft tissue was found for the first time, but was based on few cancers of assorted histology. PMID- 11024203 TI - Mortality from all cancers of asbestos factory workers in east London 1933-80. AB - OBJECTIVE: To give the observed and expected deaths due to cancer at all separate sites in asbestos workers in east London, and to analyse these for overall effect and exposure-response trend. METHODS: The mortality experience of a cohort of over 5000 men and women followed up for over 30 years since first exposure to asbestos has been extracted. RESULTS: There was a large excess of deaths due to cancer (537 observed, 222 expected). Most of these were due to cancer of the lung (232 observed, 77 expected) and pleural (52) and peritoneal (48) mesothelioma. The exposure-response trend for all these three causes was highly significant. There was also an excess of cancer of the colon (27 observed, 15 expected) which was significantly related to exposure. There were significant excesses of cancer of the ovary, of the liver, and of the oesophagus but with no consistent relation to exposure. CONCLUSIONS: The excess risk of cancer after exposure to asbestos was mainly due to cancer of the lung and mesothelioma. An exposure related excess of cancer of the colon was also detected but the possibility that some of these deaths may have been peritoneal mesotheliomas could not be excluded. There was no consistent evidence of exposure related excesses at any other site. PMID- 11024205 TI - Analysis of genotype and phenotype in fukuyama congenital muscular dystrophy. AB - OBJECTIVE: To do a genetic study of clinically diagnosed Fukuyama congenital muscular dystrophy (FCMD) by means of haplotype analysis, and to study the correlation of genotype and phenotype in FCMD. METHODS: Six closest available informative markers for each of the mapped FCMD genes were tested in 100 Japanese families with at least one affected patient. RESULTS: Seventy-seven percent of FCMD-bearing chromosomes had an ancestral founder haplotype(138-192-147-183) and 10 chromosomes had a second founder haplotype(130-201-157-183). Thirty-five(69%) typical, and 20(77%) mild, while 5(19%) severe cases were homozygous for the founder haplotype. On the contrary, 13(25%) typical, 5(19%) mild, and 15(65%) severe cases were heterozygous. Eight of 10 that were heterozygous for the second founder haplotype had the severe phenotypes. Two typical cases with severe brain and ophthalmologic anomalies were also heterozygous for the second founder haplotype. CONCLUSION: Homozygosity was associated with the typical and mild phenotypes, while compound heterozygosity was related to the severe phenotype. The value of the second founder haplotype in predicting disease severity was suggested. PMID- 11024204 TI - Fungal and other spore counts as predictors of admissions for asthma in the Trent region. AB - OBJECTIVES: The importance of airborne fungal and other spores in provoking asthma attacks is uncertain. Panel studies have generated evidence that suggests a link between outdoor spore counts and severity of asthma. There have been no population based time series studies relating outdoor exposure to spores with incidence of attacks of asthma. METHODS: Outcomes were hospital admissions for asthma on 2002 days during 1987-94, for children and adults in the Trent region of England. Predictors were daily counts of 25 spore taxa from volumetric traps in Derby on the same and previous day. Admissions for asthma were adjusted for weekly, seasonal, and longer term trends by log linear autoregressive models. Spore counts on 6 days of asthma epidemics were also examined. RESULTS: When spore counts for individual taxa were analysed as quantitative variables, two positive and two negative correlations (out of a possible 100) were significant at the 5% level. When spore counts were dichotomised at the 90th percentile, one negative and eight positive correlations (out of 100) were significant at the 5% level. All significantly positive associations related to admissions among children, but none involved the total spore count. However, total spores were above the 90th percentile on four of the six epidemic days (odds ratio (OR) 9.92, 95% confidence interval (95% CI) 1.41 to 109.84), but epidemics occurred on only four of 343 days with high total moulds. CONCLUSIONS: There was some evidence that exceptional rates of admission for asthma tend to occur on days with high total mould spore counts, but no specific taxon was consistently implicated. The predictive power was insufficient to support a public warning system. PMID- 11024206 TI - An experimental study on the developmental malformations of the trisomy 18 NMRI mice. AB - OBJECTIVE: To study the incidence of trisomy 18 in NMRI mice and its developmental malformations. METHODS: Sixty female NMRI mice and 11 male mice with the Rb(2.18) 6 Rma and Rb (1 +/-18) 10 Rma chromosomes were used. On the gestational days 16 and 17, the pregnant mice were randomly killed by cervical dislocation. A total of 823 implants were gained from 60 litters; they comprised 569 living foetuses, and chromosomal examination was performed in the generated living foetuses. Gross morphological study was performed in all living fotuses. RESULTS: Ninety-five foetuses were identified as trisomic foetuses in all of 569 living foetuses. Among them, there were 68 trisomy 18 NMRI mice with cleft palate. Gross morphological study showed that trisomy 18 foetuses had significant body flexion. Some cases of trisomy 18 showed shortened limbs, oedematous body or exencephaly as additional symptoms. CONCLUSION: Developmental malformations in trisomy 18 NMRI mice showed a seried of abnormalities; they had not only the underdeveloped body weight, but also the highter incidence of congenital cleft palate. PMID- 11024207 TI - HER2 promoter controlled specific expression of the reporter gene in ovarian cancer cell line. AB - OBJECTIVE: Construct a retrovirus vector that expresses the downstream gene specifically in ovarian cancer cells so as to increase the specificity of gene therapy. METHODS: HER2 promoter obtained by PCR was cloned into P(LXSN); mutant GFP cDNA was inserted just downstream of this promoter. Transfect SK-OV3 and MCF 7 cell lines with the new vector, P(LHGSN). RESULTS: Three days after transfection with P(LHGSN), green fluorescence was observed in SK-OV3 cells while not in MCF-7 cells. CONCLUSION: HER2 promoter can control tumor specific expression of the reporter gene in ovarian cancer cell lines. With proper modification, the vector is expected to be useful in tumor specific gene therapy of ovarian cancers. PMID- 11024208 TI - A new mutation in the connexin 32 gene was found in Charcot- Marie-Tooth disease in Chinese patients. AB - OBJECTIVE: To investigate the characteristics of gene mutations of connexin 32 exon 2 in Charcot-Marie-Tooth disease in Chinese patients. METHODS: Screening for connexin 32 gene mutation was conducted in 6 unrelated CMT1 patients without duplication and 10 unrelated CMT2 patients. Mobility shift of exon 2 was analyzed by SSCP and further confirmed by sequencing. The PCR products were cut by appropriate restricted enzyme in 50 normal controls. RESULTS: One missense mutation at nucleotides 62(G-->A) was found in a CMT1 patient. 50 normal controls were analyzed by the enzyme HaeIII and no abnormality was found. This proved that the mutation was the cause of disease. CONCLUSION: This mutation has not been reported previously. A proportion of CMTX patients may exist in the group of CMT1 patients in China. PMID- 11024209 TI - DNA analysis of a pedigree with myotonic dystrophy in Songjiang county, Shanghai. AB - OBJECTIVE: To make the molecular analysis of a pedigree with myotonic dystrophy (DM) in Songjiang county, Shanghai, and to observe the connection between CTG repeat number and clinical features. METHODS: In twenty-three individuals of a pedigree with DM, CTG trinucleotide repeat numbers located in the 3' untranslated region of a protein kinase gene (MTPK) on chromosome 19q13.2-3 were analyzed by using Long Expand TM Template PCR system. RESULTS: Four of eight clinical patients had expanded DM allele, the other four had no expanded CTG copies. Seven of eight suspicious DM cases had expanded CTG repeat numbers and were therefore genetically diagnosed as DM, and an asymptomatic individual was diagnosed as a doubted DM patient by DNA analysis. High risk of DM in six of seven individuals was ruled out, and a clinical doubted DM individual was ascertained a normal person by molecular analysis. A positive correlation was found beween early onset and expanded CTG repeat number in six parents (or grandparents)/child pairs, but in the pair II 2 /IV 7 CTG repeat numbers were reduced from 3100 in the grandmother to 175 in her grandson and there was no anticipation phenomenon. CONCLUSION: The measurement of CTG repeat number can help to ascertain the diagnosis of DM in clinical and preclinical patients, but some clinically- diagnosed DM patients might have normal CTG repeat numbers. Anticipation phenomena were common in the pedigree. PMID- 11024210 TI - Preliminary studies on parkin gene deletion at exons 1 to 6 in Chinese patients with praecox Parkinson's disease. AB - OBJECTIVE: To search parkin gene deletion mutations at exons 1 to 6 in Chinese patients with praecox Parkinson's disease (PPD) and analyze them together with the clinical features of PPD. METHODS: DNA was extracted from peripheral blood of PPD patients; deletion mutations of parkin gene were identified by PCR amplification and agarose gel electrophoresis. The clinical data were analyzed together with the above information. RESULTS: Out of 21 patients, 2 had exon 1 deletion, 2 exon 4 deletion, and 1 exon 6 deletion. The mean age of the patients with deletion mutation was 45.7+/-1.8 years. All of the patients who had parkin gene deletion mutation had tremor, rigidity and bradykinesia, but athetosis and family history of PD were not found. In additon, the deletion mutations of parkin gene at exons 2, 3, 5, were not found. CONCLUSION: There are deletion mutations of parkin gene at exons 1,4,6 in PPD of Chinese, and such deletions generally happen on middle-aged patients. PMID- 11024211 TI - 1376G-->T mutation of G6PD gene in Han and Li nationalities in Hainan, China. AB - OBJECTIVE: To investigate the 1376G-->T mutation of G6PD gene in the cases of G6PD deficiency in the Han nationality and the Li nationality of Hainan, China. METHODS: DNAs were extracted from the white blood cells of G6PD deficient cases by salt-out method. Allelic specific polymerase chain reaction was used to detect the 1376G-->T mutation. RESULTS: Fifty-nine Han nationality cases and 32 Li nationality cases were analysed; the 1376G-->T mutation was found in 19 Han cases(32.2%) and 18 Li cases(56.2%). CONCLUSION: 1376G-->T is a common mutation which causes G6PD deficiency in the Han nationality and the Li nationality in Hainan, China. Based on the phylogenetic tree of populations in China, these results indicate that the mutation might occur prior to the divergence of the Han and Li nationalities. It is of significance to study the mutations of G6PD gene in different nationalities in China for elucidating the origin, migration and evolution of the nationalities. PMID- 11024212 TI - Studies of MAL gene in human esophageal cancer by RNA in situ hybridization. AB - OBJECTIVE: To observe the expression of MAL gene mRNA in esophageal cancer tissue and adjacent epithelial mucosa. METHODS: Non-radioactive RNA in situ hybridization. RESULTS: MAL gene was expressed in all epithelia of 33 matched samples but only in 8 of esophageal cancer tissues. Differential expression between esophageal cancer tissues and adjacent epithelia was not related to clinical parameters. CONCLUSION: Down-regulation of MAL may be an important event in the development of esophageal cancer. PMID- 11024213 TI - Study on polymorphism of MICA exon 5 microsatellite in Shanghai Han population. AB - OBJECTIVE: To investigate the polymorphism distribution of MICA gene exon 5 microsatellite in Shanghai Han population and study the relationship of MICA gene and its closely linked gene HLA B. METHODS: The microsatellite polymorphism of MICA exon 5 in 175 unrelated healthy individuals were investigated using PCR heteroduplex analysis. RESULTS: (1)There are 5 alleles of the microsatellite polymorphism in exon 5 of the MICA gene in Shanghai Han population, the gene frequency of MICA*A5 is the highest(39.14%), the next is MICA*A5.1(22.29%). (2)There is strong linkage disequilibrium between the HLA-B and MICA STR loci, e.g. MICA*A5 and HLA-B15, MICA*A4 and HLA-B27. CONCLUSION: The polymorphism distribution of MICA gene and the linkage disequilibrium between MICA and HLA-B have deviations in different ethnic groups. PMID- 11024214 TI - The relationship between angiotensinogen gene CD235 met-->Thr substitution polymorphism and brain infarction in Chinese. AB - OBJECTIVE: To detect the relationship between angiotensinogen (AGT) gene CD235 Met-->Thr substitution polymorphism and brain infarction. METHODS: The CD235Met- >Thr substitution polymorphism in exon 2 of AGT gene was analyzed in normal group(82 cases) and brain infarction group (102 cases) by a combination of polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). RESULTS: Between the control group and brain infarction group, chi-square test showed no significant difference in the frequencies of T/T genotype and T allele (P>0.05). However, the frequencies of T/T genotype and T allele (70.6% and 83.8%) for multiple brain infarction patients were higher than those (42.6% and 65.2%) for lacunae brain infarction patients and those (42.0% and 65.2%) for controls (P<0.01). The frequencies of AGT T/T genotype and T allele (65.3% and 78.6%) for patients with brain infarction associated with hypertension were higher than those (42.0% and 65.0%) for patients with brain infarction not associated with hypertension (P<0.05). CONCLUSION: No direct relationship between AGT CD235 Met- >Thr substitution polymorphism and ordinary brain infarction has been observed, but there is a significant correlation between AGT gene T/T genotype and multiple brain infarction, and brain infarction associated with hypertension too. PMID- 11024216 TI - Alternative splicing of RHAMM gene in chinese gastric cancers and its in vitro regulation. AB - OBJECTIVE: To identify the receptor of hyaluronic acid mediated motility (RHAMM), expression pattern(s) in the gastric mucosa with various lesions and to elucidate the effect of hyaluronic acid(HA) on in vitro RHAMM expression. METHODS: Immunocytochemical staining and Western blot hybridization were performed to check the expression patterns and locations of RHAMM molecules in the noncancerous, preneoplastic and malignant gastric specimens. RHAMM expression of the two stomach cancer cell lines cultured conventionally or under the conditions with gelatin or HA supplementation was evaluated as well. RESULTS: Mr 93000-95000 intracellular RHAMM, so-called IHABP, proteins were detected in 17% noncancerous mucosa. The multiple spectrums and increased frequencies of RHAMM expression could be observed in the majority of the cancer cases with both intracellular and cell surface labeling. HA but not gelatin could promote membrane type RHAMM (mRHAMM) expression of stomach cancer cells in a dose associated pattern. CONCLUSION: The expression of mRHAMM variants is closely associated with tumor progression, which may be promoted and modulated by HA. PMID- 11024215 TI - The relationship of angiotensin-converting enzyme gene to essential hypertension and drug treatment in Chongqing. AB - OBJECTIVE: To measure and discuss the association of the angiotensin converting enzyme (ACE) gene insertion/deletion(I/D) polymorphism with essential hypertension and drug treatment in Chongqing. METHODS: Polymerase chain reaction(PCR) method was used to amplify the sixteenth intron of the ACE gene for the analysis of 114 healthy controls and 75 patients with essential hypertension. At the same time, 49 patients with essential hypertension were randomly divided into two groups for a placebo-controlled crossover comparison of ACEI versus CCB. RESULTS: The frequency of homozygous alleles DD and the frequency of deletion alleles were significantly higher in the essential hypertension group than in the healthy controls(P>0.005). The blood pressure decreased more in the DD genotype subjects when treated with the ACEI but decreased less when treated with the CCB (15.91 vs 12.43, P<0.1). The blood pressure decreased more in the II genotype subjects when treated with the CCB but decreased less when treated with the ACEI (16.55 vs 9.6, P<0.05). CONCLUSION: The results indicate that a deletion (DD) polymorphism of the ACE gene is associated with essential hypertension in Chongqing. Detection of the ACE genotype in clinics is helpful to the selection of antihypertensive drugs. PMID- 11024217 TI - Dopamine receptor gene polymorphisms in Guangzhou Hans. AB - OBJECTIVE: To determine the distribution of dopamine D2, D3, and D5 receptor(DRD2, DRD3, DRD5) gene polymorphisms in Guangzhou Hans. METHODS: A total of 141 healthy Guangzhou Hans were studied by the use of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and PCR amplification of specific allele (PASA) techniques. Current results were compared with the data on other ethnic groups. RESULTS: Within the 141 individuals tested, the frequencies of the A1(Taq I-) and A2(Taq I+) alleles of the TaqI A mutation site in the 3'non-coding region of the DRD2 gene were found to be 48% and 52%, respectively. The observed frequencies of the A1A1, A1A2 and A2A2 genotypes were found to be 17%, 52% and 31%, respectively, which met Hardy-Weinberg equilibrium. The frequencies of the A1 and A2 alleles of the Bal I mutation site in the exon 1 of DRD3 gene were found to be 73% and 27%, respectively. The observed frequencies of the A1A1, A1A2 and A2A2 genotypes were found to be 53%, 39% and 8%, respectively, which met Hardy-Weinberg equilibrium. The frequencies of the 1 and 2 alleles of the Msp I mutation site in the intron 5 of DRD3 gene were found to be 62.5% and 37.5%, respectively. The observed frequencies of the 1-1, 1-2 and 2 2 genotypes were found to be 35%, 55% and 10%, respectively, which met Hardy Weinberg equilibrium. No linkage disequilibrium was observed between the Bal I and Msp I polymorphism in Guangzhou Hans(chi(2)=0165, P>0.05). The frequencies of the T and C alleles of the T978C mutation site of DRD5 gene were found to be 51% and 49%, respectively. The observed frequencies of the T/T, T/C and C/C genotypes were found to be 23.6%, 54.6% and 21.8%, respectively, which met Hardy-Weinberg equilibrium. CONCLUSION: The polymorphisms of DRD2, DRD3, DRD5 gene in Guangzhou Hans were high and different from those in other populations. PMID- 11024218 TI - The application of CD71 and Hoechst33258 to staining method for sorting fetal nucleated red blood cells in the peripheral blood of pregnant women. AB - OBJECTIVE: To compare mononuclear cells labeled by CD71 single-staining method with CD71 and Hoechst33258(HO258) double-staining one, and sort nucleated red blood cells (NRBCs) in peripheral blood in pregnant women by use of the latter method. METHODS: Mononuclear cells in umbilical blood and peripheral blood in normal pregnant women and patients with mild or moderate pregnant hypertension syndrome were labeled by erythrocyte specific antibody CD71 and nuclear dye HO258. Furthermore, NRBCs were sorted in combination with the use of flow cytometry. RESULTS: Labeled by CD71 and Hoechst33258 double-staining method, the positive labeling rate of mononuclear cells in umbilical blood and peripheral blood in normal pregnant women and patients with pregnant hypertension syndrome was remarkably lower than that labeled by CD71 single-staining method (t-test, P<0.01). Meanwhile, CD71 and Hoechst33258 double-staining method was successfully used in sorting NRBCs in peripheral blood in pregnant women by flow cytometry. CONCLUSION: CD71 and HO258 double-staining method could exclude mature erythrocytes, thrombocytes and reticulocytes and this improve NRBCs' specific label and sorting purity. PMID- 11024219 TI - New research hotspots of HLA. AB - Over a long period of time, biological questions and clinical problems have been the essential motivating factors that arouse great interest in HLA and promote the enormous efforts to research HLA. The new research trends of HLA initiated by the preceding workshops of the coming 13th International Histocompatibility Workshop and Conference(IHWC), the symposium of the 12th European Histocompatibility Conference as well as the related articles were reviewed. The development and the application of DNA-based typing for HLA class I genes have been emphasized. These efforts to analyze multiple polymorphic genes are important not only for the furthurance of understanding the immunobiology of HLA, but also for the development of model systems that will be useful in future population-based epidemiological studies aimed at polygenic diseases such as cancer. Moreover, HLA molecules are broadly specific peptide receptors governig most specific immune responses in humans. A detailed understanding of their function should be an important step towards the development of a rational strategy for immune manipulations (e.g. vaccines). PMID- 11024220 TI - Progress in the molecular genetic research of multinodular goiter. AB - Multinodular goiter is a worldwide-distributed disease, but yet its pathology and genetic etiology are not clear. At present, most researches have been restrained to traditional epidemiological survey and the disease has been rarely studied at the level of molecular genetics. The pathogenesis of multinodular goiter, as is generally accepted by most researchers, can be attributed to many factors such as hormones, growth factors and the inherent functional heterogeneity of thyroid follicles. Since hormone and iodine metabolization are widely recognized as a major mechanism in determining the formation of multinodular goiter, some reports in literature are mainly focused on such genes that are responsible for hormone synthesis and iodine metabolization. Mapping experimental data were available to support location of multinodular goiter gene(s) onto chromosome 14q by whole genome scanning in a large pedigree analysis. Additional data, particularly those extracted from large scaled marker-assisted mapping experiments, are important so as to confirm the gene location, to improve resolution of the location, and finally to dissect the genes underlying the disease at molecular level. PMID- 11024221 TI - Differences between users and non-users of emergency contraception after a recognized unprotected intercourse. AB - Knowledge of emergency contraception is crucial but might not transform into use. Factors influencing decision-making related to use of emergency contraception after an unprotected intercourse and the characteristics of users of emergency contraception (EC) were assessed. In an abortion clinic setting, 217 women referred for termination of pregnancy were asked to fill in a questionnaire. Of the 217 women, 139 (64%) were aware of pregnancy risk but only 9 (4%) had used EC after the unprotected intercourse. 42% were estimated to have sufficient knowledge to use hormonal emergency contraception. In a larger background population, a calculated 29% used EC after a recognized unprotected intercourse. EC users were older, better educated, more often in stable relationships, had experienced more abortions, and gestation age was less. However, younger women were in general better informed of EC. Knowledge of EC does not necessarily transform into action. Neglect of risk after an unprotected intercourse is frequent in younger well-informed women and information has to be better targeted. PMID- 11024222 TI - Birth control method choice and use of barrier methods for sexually transmitted disease prevention among low-income African-American women. AB - In a prospective study of 1122 women attending a sexually transmitted disease (STD) clinic in Alabama, consistent use of the male condom and vaginal microbicide/spermicides was promoted to reduce STD risk. This analysis evaluated: 1) baseline characteristics that may influence birth control method choice; and 2) the association of birth control method and other baseline characteristics with consistency of barrier use during follow up. Birth control method was associated with sociodemographic variables, sexual, and reproductive history. Women who adopted user-independent methods (tubal ligation, implants, injectable hormones) appear to have completed their family plan. Oral contraceptive users were of higher socioeconomic status and at lower STD risk. Barrier method users and women who used no method were young and at higher STD risk. Consistency of condom/spermicide use increased in all groups. Barrier method users were more likely than other women to use condoms and spermicides during the study. Women who used no birth control method at baseline experienced the largest increase in barrier use during follow up, although their barrier use rates were lower than in other groups. The synergism between the intention to prevent pregnancy and the intention to prevent STD should be considered in the design of interventions promoting condom use. PMID- 11024223 TI - A multicentered clinical trial of the long-acting injectable contraceptive Depo Provera in Chinese women. AB - This study was an open trial where 1994 subjects each received Depo Provera injectable contraceptive every three months for one year and were observed a total of 20,294.3 woman months. At the time of observation, only one accidental pregnancy had occurred giving a use-effectiveness rate of 99.94% and a cumulative continuation rate of 72.87%. There were no significant adverse effects on weight and blood pressure. The main side effects were spotting, prolonged bleeding, and amenorrhea and these were also the main reasons of discontinuation. Complaints related to bleeding problems gradually decreased and complaints of amenorrhea increased with continued use. In lactating women, side effects occurrence rate was lower and continuation rate was higher in comparison with non-lactating women. Users did not report any effect on milk secretion. The results of this study confirm that Depo Provera is a very effective contraceptive method. With appropriate counseling and medical support, high acceptability and continuation rate can be achieved. Depo Provera is especially suitable for lactating women and could become a popular contraceptive method for lactating Chinese women. PMID- 11024224 TI - Glucose metabolism among Norplant users in northern Nigeria. AB - Twenty-one women who were consecutive acceptors of Norplant were recruited for this study in our centre in August 1997. The patients had baseline (pre insertion) investigations including a standard oral glucose tolerance test (OGTT). At their regular 3, 6, and 12 months follow-up visits, the OGTT was repeated and results were analyzed. The mean age of the clients was 31.3+/-4.51 years (range 24-40 years). The mean weights were 61.60+/-11.35, 57.63+/-7.51, 62.60+/-11.98, and 62.17+/-11.56 kg at pre-insertion, 3, 6, and 12 months follow up visits respectively. The differences were statistically insignificant. All OGTT values at pre-insertion and at the follow-up visits were within the normal range. However, in comparison to pre-insertion levels (4.00+/-0.10 mmol/L), the fasting blood glucose concentration at 12 months (3.39+/-0.12 mmol/L) showed a statistically significant decline (p <0.001). The area under the glucose curve (AUC) rose by 5.8% (p = 0.018) at 3 months but declined by 5.8% (p = 0.110) and 7.3% (p = 0.103) at 6 and 12 months, respectively. The peak pre-insertion plasma glucose level following OGTT occurred at 30 min. At 3 months, the peak OGTT glucose level occurred at 30 minutes and plateaued until 60 min while at 6 and 12 months this occurred at 60 and 90 minutes, respectively. The overall trend of OGTT results within the study period showed that 12 months of Norplant use in Northern Nigerian women has no detrimental effect on glucose metabolism. PMID- 11024225 TI - The effect upon the human vaginal histology of the long-term use of the injectable contraceptive Depo-Provera. AB - The objective of the study was to evaluate the effect of long-term use of the injectable contraceptive depot medroxyprogesterone acetate (DMPA) on human vaginal histology. Twenty premenopausal women currently using DMPA as a contraceptive method for two and three years were compared with 20 regularly menstruating women, who never used Depo-Provera and/or other kind of hormonal contraceptive in the last 6 months prior to the study. Subjects and controls were matched by age (+/-1 year), body mass index (kg/m2) (+/-1.0), number of pregnancies (+/-1), age at first intercourse (+/-1 year), years of sexual activity (+/-1 year), and number of partners during their life (+/-1). Vaginal biopsies were performed in users at 90+/-7 days after the last injection and in nonusers at day 20-25 of the menstrual cycle. In addition, at the day of the biopsy a blood sample was collected to measure estradiol (in all women) and DMPA in users. The level of serum estradiol was significant lower in Depo-Provera users than in controls (p < 0.001). The thickness of the vaginal epithelium was not smaller among DMPA users than among controls, the mean count of Langerhans cells per mm of epithelium were almost identical in both groups, and no significant differences were found on the vaginal maturation indices. In conclusion, the use of Depo-Provera between two and three years did not affect vaginal thinning of the epithelium, Langerhans cell count or maturation index. PMID- 11024227 TI - Anti-nidatory effect of vaginally administered (Ala8,13, 18)-magainin II amide in the rhesus monkey. AB - The hypothesis that timed application of a potent anti-microbial, anti-tumor agent like magainin peptides can inhibit blastocyst implantation was examined in the present study using the rhesus monkey as the primate model. Incidence of pregnancy, vaginal bleeding patterns, serum levels of progesterone, estrogen and monkey chorionic gonadotropin were examined following vaginal administration of (Ala8,13,18)-magainin II amide, a synthetic analogue of magainin 2, via tampon during days 20 to 26 of mated cycles. Implantation occurred in two out of three animals following administration of 0.25 mg magainin, while administration of 0.5 mg (Ala8,13,18)-magainin II amide resulted in inhibition of implantation in all females with no change in lengths of treatment cycles, and subsequent cycles. It appears from the present study that, besides being a local microbicidal agent, intravaginal administration of (Ala(8,13,18))-magainin II amide is a potential anti-implantation strategy for intercepting pregnancy. PMID- 11024226 TI - Drospirenone: pharmacology and pharmacokinetics of a unique progestogen. AB - The pharmacology and pharmacokinetics of drospirenone, a unique progestogen, are reviewed in this paper. Unlike other progestogens, drospirenone, an analogue of spironolactone, has biochemical and pharmacologic profiles similar to endogenous progesterone, especially regarding antimineralocorticoid and antiandrogenic activities. Drospirenone counteracts the estrogen-induced stimulation of the renin-angiotensin-aldosterone system and blocks testosterone from binding to androgen receptors. Because of these characteristics, it has the potential to reduce body weight, blood pressure, and low-density lipoprotein levels and to enhance high-density lipoprotein levels. As a combination oral contraceptive, drospirenone with ethinyl estradiol is effective and has positive effects on weight and lipid levels. Additionally, it relieves menstrually related symptoms (e.g., negative affect and water retention) that are commonly observed with other combination oral contraceptives. Based on the biochemical and pharmacodynamic data, drospirenone appears to be a viable alternative to the currently available progestogens. PMID- 11024228 TI - Decrease in sperm number after treatment of rats with Austroplenckia populnea. AB - The purpose of this study was to evaluate the effects of a hexanic extract (HE) made from leaves of A. populnea collected in Botucatu, State of Sao Paulo, and Nova Lima, State of Minas Gerais, Brazil, at a range of doses during 7 and 14 days, on the male reproductive system of rats. The treatment did not affect the body weight, nor absolute organ weight. The serum testosterone levels, testicular sperm head counts, daily sperm production, and sperm morphology did not differ from that of the control groups. The spermatogenesis and the morphometric parameters of cauda epididymidis were not affected by the treatment. Cauda epididymis sperm number was significantly reduced in the group that received HE of Nova Lima, 1 g/kg/day, during 14 days, from the control group. PMID- 11024230 TI - On the first try? PMID- 11024229 TI - Modulation of morphological changes of endometrial surface epithelium by administration of composite root extract in albino rat. AB - In the present investigation, we attempted to study the modulation of morphological changes of endometrial surface epithelium by Scanning Electron Microscope (SCEM) in the adult albino rat after oral administration of roots extract. The origin of this experiment lies in the fact that the dry powdered roots of five plants, i.e., Plumbago rosea, Borassus flabellifer, Carica papaya (male), Dolichos lablab, and Shorea robusta in mixed form, has been traditionally used by the folk women of this region through oral route during the first seven days of a menstrual cycle to prevent conception. Ethanolic crude extract of these roots in composite form which has been referred to here as composite root extract (CRE) was administered to the adult female albino rat in a dose of 1000 mg/kg body weight/day through oral route continuously for 12 days starting from proestrous. The observations under SCEM showed that administration of the CRE induced morphological changes of the endometrial surface epithelium in albino rat uterus. The characteristic smooth regular pattern of normal epithelium appears to have been changed at places by haphazardly oriented groups of cells and loss of microvilli indicating a disorganized picture. This structural disparity has been thought to affect the smooth functioning of nidatory preparation in the endometrium. PMID- 11024231 TI - Surgery for constipation update(1). PMID- 11024232 TI - Tumor implantation during laparoscopy: is it really a problem?(1). PMID- 11024233 TI - Neuroblastoma: prognostic indicators(1). PMID- 11024234 TI - Liposuction: fact, fiction, and future(1). PMID- 11024235 TI - Trends in the surgical management of gastric cancer(1). PMID- 11024236 TI - Helicobacter pylori update(1). PMID- 11024237 TI - Open anterior repair of inguinal hernias with mesh(1). PMID- 11024238 TI - E-mail tips, tricks, and trouble spots: part 2. PMID- 11024239 TI - Introduction. PMID- 11024241 TI - Laparoscopic localization and management of lower gastrointestinal vascular lesions. PMID- 11024240 TI - Lower gastrointestinal vascular lesions: current concepts in diagnosis and treatment. PMID- 11024242 TI - Grand rounds in general surgery. PMID- 11024243 TI - Research funding for surgical residents. PMID- 11024244 TI - Society of Air Force Clinical Surgeons' 47th Annual Symposium. PMID- 11024245 TI - The Robert E. Lee Memorial Lecture, April 4, 2000: history of the Society of Air Force Clinical Surgeons. PMID- 11024246 TI - A 5-year review of abdominal aortic aneurysms at a VA medical center. AB - We sought to determine the outcome of our policy of following asymptomatic abdominal aortic aneursyms until a maximal diameter of 5 cm is reached before assessing risk factors and need for operative intervention.We retrospectively reviewed the charts of all patients identified with abdominal aortic aneurysm in our hospital system by ICD code for the past 5 years. We specifically reviewed size of aneurysm, modality of assessment (computed tomography vs ultrasonography), age of patient, whether aneurysm repair had been performed, comorbidities, and current status (living or deceased).Eighty-eight patients were identified with abdominal aortic aneursyms. No ruptured aneurysms were identified in any of the patients that have been followed, regardless of size. All patients who have undergone an aneurysmorrhaphy are currently alive.Our current policy of following abdominal aortic aneursyms, as stated above, has not resulted in either ruptured aneurysm or postoperative death, and we feel that in the veteran population that this is a sound approach. (Curr Surg 57:343-345) PMID- 11024247 TI - Black widow spider (Latrodectus mactans) envenomation in a term pregnancy. AB - Description of a black widow spider (Latrodectus mactans) envenomation in a term pregnancy.Case report conducted at an Air Force tertiary care hospital of a 27 year-old primigravida at 38 1/7 weeks pregnancy.Latrodectus mactans antivenin can be given to treat symptoms of black widow envenomation.Black widow envenomations can cause symptoms associated with acute intra-abdominal processes. In pregnancy, envenomations can result in symptoms and signs similar to those seen in preeclampsia (abdominal pain, headache, hypertension, and proteinuria). Latrodectism should be considered in patients complaining of these symptoms in association with a spider bite. If latrodectism is considered to be the underlying origin for these symptoms, appropriate treatment should be administered. In cases of pregnancy, treatment should include L. mactans antivenin if believed to be clinically indicated. There is no current evidence that this antivenin is contraindicated in pregnancy. (Curr Surg 57:346-348) PMID- 11024248 TI - Can one lung ventilation prevent air embolism in the lung injury victim? AB - The current literature indicates that patients with hilar lung injury who are receiving positive pressure ventilation are at risk for systemic air embolism, but no studies have yet tested an alternative to the current management: immediate thoracotomy and hilar clamping. We wanted to demonstrate that one lung ventilation of the uninjured lung protects against the formation of arterial air embolism in the presence of contralateral hilar lung injury.In 6 juvenile swine, the right bronchus was selectively ventilated, and ultrasound of the abdominal aorta was used to detect air emboli. The hilum of the left lung was stabbed with a scalpel; after a brief period of monitoring to detect air emboli, the tip of the endotracheal tube was withdrawn into the trachea and the left lung ventilated.Air emboli were detected in 2 animals. The air emboli did not form while the lung was isolated, but they did appear immediately when the endotracheal tube was withdrawn into the trachea. Air was also noted in the chambers of the heart and coronary arteries, and led to fibrillation and death.One lung ventilation appears to protect against arterial air embolism in unilateral hilar lung injury. (Curr Surg 57:349-353) PMID- 11024249 TI - Isolated subaxial cervical spine stenosis in achondroplasia. AB - Although isolated subaxial cervical spine stenosis in achondroplasia is less common than narrowing at the foramen magnum, thoracolumbar junction, or lumbar spine, it should be recognized as a distinct entity. These congenital changes usually become symptomatic when combined with degenerative changes in early adult life. Yearly monitoring of adult achondroplastic patients for signs and symptoms of cervical myelopathy or radiculopathy is recommended. When this develops, early surgical decompression should be considered, as these patients generally do not respond well to nonoperative treatment. The altered anatomy and small size make these cases particularly challenging. A thorough history and examination and complete diagnostic testing are necessary to gain an understanding of the exact etiology of the symptoms. Wide laminectomies and foraminotomies are then usually required to decompress the spinal cord and nerve roots. Fusion using internal fixation may be required to stabilize the spine if excessive bone is removed. (Curr Surg 57:354-356) PMID- 11024250 TI - Myxoma involving the atrial valve chordae. AB - Primary cardiac valve tumors are rare and comprise less than 10% of all cardiac tumors. They are, however, of clinical importance because of their unique locations. We report an unusual case of myxoma involving the tricuspid valve chordae in a young woman presenting with syncope. The myxoma was diagnosed by echocardiography and successfully removed by excision of the involved tricuspid valve chordae with valve preservation. (Curr Surg 57:357-358) PMID- 11024251 TI - The effects of TRICARE implementation on general surgery residency training at Travis Air Force Base. AB - TRICARE is the military version of health care insurance. The conversion from CHAMPUS (the former version) to TRICARE was completed at David Grant Medical Center (DGMC) in October 1995. Because of a perceived decrease in the overall surgical caseload at DGMC in recent years, a review of the operative case logs of all graduating chief residents in general surgery was undertaken. The goal was to determine if institution of TRICARE was adversely affecting the surgical experience of our residents.A retrospective review of the residency review committee (RRC) case logs of all graduating chief residents in general surgery at David Grant Medical Center was performed from 1990 to 1999. The review was divided into 2 groups: group 1 included all individuals who graduated in those years before initiation of TRICARE (1990 to 1995); group 2 included those who graduated after the initiation of TRICARE (1996 to 1999). Each of the 16 separate RRC reporting categories and 3 summary categories were compared to detect differences in overall caseloads between group 1 and group 2.Compared with group 1, group 2 had significantly fewer aortic, pediatric surgery, endoscopy, and total cases done in the chief year.Since the initiation of the TRICARE health plan at David Grant Medical Center, overall caseloads have not declined. However, there has been a concerning decline in chief resident-year cases. This decline may not become fully apparent for several more years as the current junior residents report their caseloads upon their graduation. (Curr Surg 57:359-362) PMID- 11024252 TI - Presidential address: the education of the surgeon in the 21st century. PMID- 11024253 TI - Experience and visual perception in resident acquisition of laparoscopic skills. AB - Assess the role of experience and visual perception (VP) in resident acquisition of laparoscopic surgical skills (LSS).Thrity-nine residents (20 PGY-1s tested just before starting residency; 19 PGY 3+) completed an LSS course, including examination of course-specific knowledge before and after didactic tutorials, 10 trials of 3 dexterity drills and suturing, and 3 standardized VP tests.Mean speed increased significantly (p < 0.001) across trials for all dexterity drills and suturing. Senior residents performed suturing trials 1 to 4 significantly faster (p < 0.05) than did PGY 1's (M +/- SD averaged across trials 1 to 4: 166.5 +/- 59.9 vs 252.3 +/- 108.2 seconds, p < 0.01). Group differences on later trials were progressively smaller and nonsignificant, as were all group differences on dexterity drills. Significant correlations between VP and speed on drills ranged from (r = -0.41, p < 0.01) to (r = -0.71, p < 0.001). Visual perception did not correlate significantly with suturing speed; neither pretest nor posttest scores correlated significantly with drill or suturing speeds.Residents at all levels can significantly increase LSS performance speed to comparable levels during a brief intensive skills course. Proficiency in specific aspects of VP is directly associated with performance speed on dexterity drills, shown in previous studies to be fundamental in the development of intracorporeal suturing skill. (Curr Surg 57:368-372. Copyright 2000 by the Association of Program Directors in Surgery.) PMID- 11024254 TI - Lemonade from lemons: a program response to RRC-determined probation. AB - To assess resident satisfaction as a result of changes made to an integrated surgical residency in response to probation.The University of Connecticut Integrated General Surgery Residency (UCIGSR), which consists of 5 hospitals, 18 rotations, and has a complement of 44 residents, was placed on probation by the Residency Review Committee (RRC) in Surgery in November 1998. Among the deficiencies cited by the RRC were inadequate evaluation of the program, rotations, residents, and the faculty by the residents, along with inadequate documentation issues. Lack of organized educational conferences, lack of faculty involvement, excessive service responsibilities and work hours, and hospital environmental issues were also cited. It was also apparent that morale of the residents and faculty had significantly eroded. Rather than trying patchwork "fixes" of deficiencies, we used this opportunity to reengineer the program, rededicating ourselves to educating and developing future surgeons. Task forces, including joint faculty and resident participation, were set up to develop solutions, based on the answer to the question: "Do we want a residency or not?" This focus was especially helpful to create hospital administration and faculty support.A survey was created to assess the changes made in the program. The survey included 65 questions pertaining to the learning environment, hospital and departmental support, and balance between clinical and educational responsibilities and overall working environment. Each resident was asked to fill out 3 identical surveys, 1 just before the RRC report, and the other 2 at 6-month intervals after the probation announcement. A 4-point grading scale was used.Our results were reflective of resident perceptions of improvement and increased satisfaction. These data demonstrated that the changes implemented as a result of the RRC findings had a positive effect on the residency program, and they corresponded to the areas where changes were implemented. This survey also gave us a method to evaluate ongoing changes in our residency.Probation can be a potent stimulus for improvement of a surgical residency. (Curr Surg 57:373-376. Copyright 2000 by the Association of Program Directors in Surgery.) PMID- 11024255 TI - Problem-based anatomy for surgical residents. AB - Graduating medical students who enter surgery residencies frequently have limited anatomic experiences during their undergraduate training. Throughout the 5 years of surgical training residents are exposed to the anatomy as it relates to the operative procedures performed. Frequently, however, many of the complex anatomic areas of the body escape the experience of a surgery resident in training. The American Board of Surgery requires that surgeons have comprehensive experience with problems located in the head and neck, chest, abdomen, pelvis, and extremities. In an effort to provide an experience in the critical anatomic areas, the Michigan State University (MSU) Integrated General Surgery Residency developed a problem-based anatomy course that focuses on some of the most difficult anatomic areas in which surgeons are expected to provide lifesaving procedures.A program was developed that would present a series of clinical scenarios focusing on critical anatomic approaches and related structures in the head and neck, thorax, abdomen, pelvis, perineum, upper extremity, and lower extremity. The program involved 61 individual clinical scenarios, each requiring specific dissections. The course is directed by a surgeon and anatomist, with the discussion focused entirely on surgical approaches, related anatomic structures, and the options of therapy. The course has been taught for 4 years with great success. The program is now being extended to all affiliated general surgery residencies in the MSU system.A problem-based anatomy course has been developed that addresses many of the complex anatomic areas in which surgeons are expected to provide safe operative intervention. A problem-based approach has provided opportunities for surgical residents to consider operative approaches necessary to perform procedures in a safe, competent manner. The success of this approach would lend credence to expanding its utility to other specialties and even medical student training. (Curr Surg 57:377-380. Copyright 2000 by the Association of Program Directors in Surgery.) PMID- 11024256 TI - The importance of vascular surgery training in a general surgery practice. AB - The aim of this report was to document the practice of vascular surgery for graduates of 4 general surgery training programs.Graduating residents from 1991 to 1995 were surveyed by phone to document the number of reconstructive vascular and dialysis access procedures that they performed during the most recent 12 months. Those who pursued additional training beyond general surgery or who did not successfully complete the certifying examination of the American Board of Surgery (ABS) were excluded. In addition, the Resident Review Committee for Surgery (RRC-S) defined category report for these same general surgeons during their residency was examined.Fifty-five percent (26 of 47) of the board-certified general surgeons do either reconstructive or dialysis access vascular surgery. The average number of procedures in the RRC-S defined category for this cohort was 76. During the focused 12 months, 1986 vascular procedures were done by these 26 surgeons (76 cases per surgeon).In certain regions of this country, a significant volume of vascular surgery is done by general surgeons who have an ABS primary certificate alone. The technique of control and repair of major arteries and veins, the consequences of distal organ ischemia, reperfusion injury, thrombosis, and embolization are important anatomic and physiologic principles that must be taught in the curriculum to general surgeons, regardless of their future surgical careers. (Curr Surg 57:381-383. Copyright 2000 by the Association of Program Directors in Surgery.) PMID- 11024257 TI - Using prospective outcomes data to improve morbidity and mortality conferences. AB - Background:Though a traditional part of most training programs, surgical morbidity and mortality (M&M) conferences are not optimal for teaching residents how to understand and improve patient outcomes. They tend to focus on unusual rather than common problems and review complications as singular phenomena, rather than recurrent events related to specific processes of care. For these reasons, we began incorporating data from our general surgery outcomes registry into our M&M conference.Developed for both clinical research and quality improvement purposes, the outcomes registry contains prospective information about all patients undergoing general surgery procedures in the operating room (approximately 2000 per year). All adverse events occurring within 30 days of surgery are categorized, using explicit criteria, by clinical nurse coordinators. Individual complications are then collapsed into four severity grades (from a previously validated grading system), from I (not life-threatening, low complexity therapy; eg, superficial wound infection) to IV (death).Application To M&M Conferences:Before each conference the data manager supplies the responsible senior resident with information about caseloads and adverse events for the preceding month. At the conference, individual cases are presented in the context of the department's broader experience with that procedure (eg, rates of wound complications after bowel surgery over the last 2 years). In reviewing trends in complication rates over time, we also explore potential relationships between practice changes and outcomes. When appropriate, local performance is compared to external "benchmarks" using data from published studies.Incorporating prospective outcome data into the M&M conference is both feasible and practical. In addition to its educational value for both resident and attending physicians, we believe this approach creates many opportunities for improving the quality of our surgical practice. PMID- 11024258 TI - The expansion of mechanistic and organismic diversity associated with non ribosomal peptides. AB - Non-ribosomal peptides are a group of secondary metabolites with a wide range of bioactivities, produced by prokaryotes and lower eukaryotes. Recently, non ribosomal synthesis has been detected in diverse microorganisms, including the myxobacteria and cyanobacteria. Peptides biosynthesized non-ribosomally may often play a primary or secondary role in the producing organism. Non-ribosomal peptides are often small in size and contain unusual or modified amino acids. Biosynthesis occurs via large modular enzyme complexes, with each module responsible for the activation and thiolation of each amino acid, followed by peptide bond formation between activated amino acids. Modules may also be responsible for the enzymatic modification of the substrate amino acid. Recent analysis of biosynthetic gene clusters has identified novel integrated, mixed and hybrid enzyme systems. These diverse mechanisms of biosynthesis result in the wide variety of non-ribosomal peptide structures and bioactivities seen today. Knowledge of these biosynthetic systems is rapidly increasing and methods of genetically engineering these systems are being developed. In the future, this may lead to rational drug design through combinatorial biosynthesis of these enzyme systems. PMID- 11024259 TI - Conserved cytoplasmic motifs that distinguish sub-groups of the polyprenol phosphate:N-acetylhexosamine-1-phosphate transferase family. AB - WecA, MraY and WbcO are conserved members of the polyprenol phosphate:N acetylhexosamine-1-phosphate transferase family involved in the assembly of bacterial cell walls, and catalyze reactions involving a membrane-associated polyprenol phosphate acceptor substrate and a cytoplasmically located UDP-D-amino sugar donor. MraY, WbcO and WecA purportedly utilize different UDP-sugars, although the molecular basis of this specificity is largely unknown. However, domain variations involved in specificity are predicted to occur on the cytoplasmic side of the membrane, adjacent to conserved domains involved in the mechanistic activity, and with access to the cytoplasmically located sugar nucleotides. Conserved C-terminal domains have been identified that satisfy these criteria. Topological analyses indicate that they form the highly basic, fifth cytoplasmic loop between transmembrane regions IX and X. Four diverse loops are apparent, for MraY, WecA, WbcO and RgpG, that uniquely characterize these sub groups of the transferase family, and a correlation is evident with the known or implied UDP-sugar specificity. PMID- 11024260 TI - Co-expression of the Bordetella pertussis leader peptidase I results in enhanced processing and expression of the pertussis toxin S1 subunit in Escherichia coli. AB - Bordetella pertussis is the causative agent of whooping cough. Traditional vaccines against this disease are inherently reactogenic, thus research is currently focussed on the production of less reactive, acellular vaccines. Expression of candidate antigens for these vaccines in Escherichia coli would be preferable, however, several B. pertussis antigens undergo incorrect post translational processing in E. coli. The leader peptidase gene (lep) of B. pertussis encodes a protein of 294 amino acid residues that shares homology with other prokaryote leader peptidase I sequences. Hydrophilicity analysis based on the predicted amino acid sequence has demonstrated a similar membrane topology to that of E. coli and Salmonella typhimurium leader peptidase I. Co-expression of the B. pertussis lep gene in E. coli strain TOPP2 expressing the pertussis toxin S1 subunit was found to markedly increase the expression and post-translational processing of the S1 protein. PMID- 11024261 TI - Heat shock proteins and inflammatory acne vulgaris: molecular cloning, overexpression and purification of a propionibacterium acnes GroEL and DnaK homologue. AB - Propionibacterium acnes is associated with inflammatory acne. The genes encoding two putative mediators of inflammation, the heat shock proteins GroEL and DnaK, were cloned from this organism and sequenced. groEL and dnaK encode proteins of 56.8 and 66.4 kDa, respectively, which show a high degree of homology (>75% similarity) to the GroEL and DnaK proteins of mycobacteria and streptomycetes. The promoter regions of both genes contain inverted repeat sequences believed to be involved in the transcriptional regulation of heat shock genes. Recombinant P. acnes GroEL and DnaK were overexpressed in Escherichia coli with C-terminal histidine tags. The recombinant proteins were purified from E. coli by metal affinity chromatography. These proteins will now be used in immunological investigations to determine their role in inflammatory acne. PMID- 11024262 TI - In situ assay for identifying inhibitors of bacterial transglycosylase. AB - An in situ transglycosylase assay has been developed using endogenously synthesized lipid II. The assay involves the preferential synthesis and accumulation of lipid II in a reaction mixture containing the cell wall membrane material isolated from Escherichia coli, exogenously supplied UDP-MurNAc pentapeptide, and radiolabeled UDP-GlcNAc. In the presence of Triton X-100, the radiolabeled product formed is almost exclusively lipid II, while the subsequent formation of peptidoglycan is inhibited. Removal of the detergent resulted in the synthesis of peptidoglycan (25% incorporation of radiolabeled material) from the accumulated lipid II. This reaction was inhibited by moenomycin, a known transglycosylase inhibitor. In addition, tunicamycin, which affects an earlier step of the pathway by inhibiting MraY, had no effect on the formation of peptidoglycan in this assay, as expected. Similarly, ampicillin and bacitracin did not inhibit the formation of peptidoglycan under the conditions established. PMID- 11024264 TI - A mutagenic PCR identifies isolates of Borrelia garinii responsible for Lyme borreliosis. AB - Borrelia garinii is one of the three major Borreliae responsible for Lyme borreliosis in Europe. We have characterized a protein of B. garinii (VS102) and a genomic fragment from the gene encoding this protein was cloned. The DNA sequence of the fragment showed high homology with a known gene of B. burgdorferi sensu stricto. The protein encoded by this gene in B. burgdorferi sensu stricto is a phosphocarrier protein (histidine-containing protein). A mutation T to G polymorphism at codon 57 was found to be specific to B. garinii. A PCR-based approach that allows the rapid detection of this mutation made it possible to specifically discriminate B. garinii from other B. burgdorferi genospecies with high sensitivity and specificity. PMID- 11024263 TI - Structural and functional analysis of the riboflavin synthesis genes encoding GTP cyclohydrolase II (ribA), DHBP synthase (ribBA), riboflavin synthase (ribC), and riboflavin deaminase/reductase (ribD) from Helicobacter pylori strain P1. AB - The functions of the riboflavin synthesis gene homologues ribA, ribBA, ribC, and ribD from Helicobacter pylori strain P1 were confirmed by complementation of defined Escherichia coli mutant strains. The H. pylori ribBA gene, which is similar to bifunctional ribBA genes of Gram-positive bacteria, fully complemented the ribB mutation and partially restored growth in a ribC mutant. However, ribBA did not complement the ribA mutation in E. coli, thus explaining the presence of the additional separate copy of the ribA gene in the H. pylori chromosome. In E. coli exclusively ribA conferred hemolytic activity and gave rise to production of molecules with fluorescence characteristics similar to flavins, as observed earlier. The E. coli hemolysin ClyA was not involved in causing the hemolytic phenotype. No riboflavin synthesis genes on plasmids conferred iron uptake functions to a siderophore-deficient mutant of E. coli. Marker exchange mutagenesis of the genes in H. pylori was not successful indicating that riboflavin synthesis is essential for basic metabolic functions of the gastric pathogen. PMID- 11024265 TI - A chromosomal region of Mycoplasma agalactiae containing vsp-related genes undergoes in vivo rearrangement in naturally infected animals. AB - A Mycoplasma agalactiae genomic fragment carrying four vsp-related genes (designated avg: agalactiae variable genes) was cloned, sequenced and compared to the vspA gene of Mycoplasma bovis. The following features were revealed: (i) the presence of a highly conserved vsp 5' upstream region; (ii) a highly homologous vsp N-terminal end encoding a putative lipoprotein signal sequence; (iii) sequence divergence of the rest of the mature proteins. By using avg specific probes in Southern blot analysis of genomic DNAs of M. agalactiae strains as well as of isolates from infected animals, marked DNA polymorphism of avg fragments was demonstrated. In addition, the avg genomic fingerprints were monitored for a period of 7 months, in isolates of M. agalactiae from an individual chronically infected animal. The results provided evidence that a chromosomal region of M. agalactiae, carrying vsp-related genes, undergoes rearrangements in vivo in the natural animal host during the course of infection. PMID- 11024266 TI - Antisense expression of the calmodulin gene from Colletotrichum trifolii impairs prepenetration development(1). AB - Calmodulin is a ubiquitous highly conserved calcium binding protein involved in cell signalling. Previous studies in our laboratory suggested a role for calmodulin in prepenetration morphogenesis in Colletotrichum trifolii, the causal agent of alfalfa anthracnose. In this report, we describe the cloning, sequencing and partial characterization of the calmodulin gene from C. trifolii. The gene is present as a single copy in the genome of C. trifolii and its predicted amino acid sequence shows considerable homology to other fungal calmodulins. The gene is most highly expressed during conidial germination and appressorial development. Using a Neurospora crassa inducible promoter driving the calmodulin gene in antisense orientation, transformants were obtained with constitutive levels of antisense calmodulin expression. Upon induction, transformants did not develop appressoria and were not pathogenic on alfalfa plants. PMID- 11024267 TI - Evidence for intermolecular interaction as a necessary step for pore-formation activity and toxicity of Bacillus thuringiensis Cry1Ab toxin. AB - Based on the observation of large conductance states formed by Bacillus thuringiensis Cry toxins in synthetic planar lipid bilayers and the estimation of a pore size of 10-20 A, it has been proposed that the pore could be formed by an oligomer containing four to six Cry toxin monomers. However, there is a lack of information regarding the insertion of Cry toxins into the membrane and oligomer formation. Here we provide direct evidence showing that the intermolecular interaction between Cry1Ab toxin monomers is a necessary step for pore formation and toxicity. Two Cry1Ab mutant proteins affected in different steps of their mode of action (F371A in receptor binding and H168F in pore formation) were affected in toxicity against Manduca sexta larvae. Binding analysis showed that F371A protein bound more efficiently to M. sexta brush border membrane vesicles when mixed with H168F in a one to one ratio. These mutant proteins also recovered pore-formation activity, measured with a fluorescent dye with isolated brush border membrane vesicles, and toxicity against M. sexta larvae when mixed, showing that monomers affected in different steps of their mode of action can form functional hetero-oligomers. PMID- 11024268 TI - Regulation of anaerobic arginine catabolism in Bacillus licheniformis by a protein of the Crp/Fnr family. AB - Arginine anaerobic catabolism occurs in Bacillus licheniformis through the arginine deiminase pathway, encoded by the gene cluster arcABDC. We report here the involvement of a new protein, ArcR, in the regulation of the pathway. ArcR is a protein of the Crp/Fnr family encoded by a gene located 109 bp downstream from arcC. It binds to a palindromic sequence, very similar to an Escherichia coli Crp binding site, located upstream from arcA. Residues in the C-terminal domain of Crp that form the DNA binding motif, in particular residues Arg-180 and Glu-181 that make specific bonds with DNA, are conserved in ArcR, suggesting that the complexes formed with DNA by Crp and ArcR are similar. Moreover, the pattern of DNase I hypersensitivity sites induced by the binding of ArcR suggests that ArcR bends the DNA in the same way as Crp. From the absence of anaerobic induction following inactivation of arcR and from the existence of a binding site upstream of the arcA transcription start point, it can be inferred that ArcR is an activator of the arginine deiminase pathway. PMID- 11024269 TI - The NAD-glycohydrolase (nga) gene of Streptococcus pyogenes. AB - The gene for NAD-glycohydrolase (nga) of group A streptococci (Streptococcus pyogenes) was identified and shown to be located immediately adjacent to the gene for streptolysin O (slo). The nga gene contains 1341 base pairs and encodes a protein of 447 amino acids, including an N-terminal signal peptide. Results from analysis with the polymerase chain reaction indicated that the nga gene is present in all of the strains tested. Functional extracellular NAD glycohydrolase, also known as NADase, was detected among a wide variety of clinical isolates and known laboratory strains and shown to be present in 72% of 100 strains examined. In contrast, 92% of strains isolated from patients with invasive streptococcal infections were positive for NADase production. PMID- 11024270 TI - Genetic analysis of Carboxydothermus hydrogenoformans carbon monoxide dehydrogenase genes cooF and cooS. AB - Carboxydothermus hydrogenoformans is an extremely thermophilic, Gram-positive bacterium growing on carbon monoxide (CO) as single carbon and energy source and producing only H(2) and CO(2). Carbon monoxide dehydrogenase is a key enzyme for CO metabolism. The carbon monoxide dehydrogenase genes cooF and cooS from C. hydrogenoformans were cloned and sequenced. These genes showed the highest similarity to the cooF genes from the archaeon Archaeoglobus fulgidus and the cooS gene from the bacterium Rhodospirillum rubrum, respectively. The cooS gene was identified immediately downstream of cooF, however, the cooF and cooS genes from C. hydrogenoformans have substantially different codon usage, and the cooF gene Arg codon usage pattern, dominated by AGA and AGG, resembles the archaeal pattern. The data therefore suggest lateral transfer of these genes, possibly from different donor species. PMID- 11024271 TI - Biolistic co-transformation of Metarhizium anisopliae var. acridum strain CG423 with green fluorescent protein and resistance to glufosinate ammonium. AB - Metarhizium anisopliae var. acridum (syn. M. flavoviride) is recognized as a highly specific and virulent mycopathogen of locusts and grasshoppers and is currently being developed as a biological control agent for this group of insects in Brazil. Intact conidia of M. anisopliae var. acridum strain CG423 were transformed using microparticle bombardment. Plasmids used were: (1) pBARKS1 carrying the bar gene of Streptomyces hygroscopicus fused to the Aspergillus nidulans trpC promoter, encoding resistance to glufosinate ammonium (or phosphinothricin) and modified by addition of the telomeric repeat (TTAGGG)(18) of Fusarium oxysporum and 2.pEGFP/gpd/tel carrying a red-shifted variant gene for Aequorea victoria green fluorescent protein (EGFP) which we have fused to the A. nidulans gpd promoter and trpC terminator. Highly fluorescent co-transformants were selected on solid minimal medium containing 100 microg ml(-1) glufosinate ammonium using an inverted microscope with 450-490 nm excitation/510 nm emission filter set. Southern blot analysis of co-transformants revealed varying multiple chromosomal integrations of both bar and egfp genes at both telomeric and non telomeric loci. Transformants retained pathogenicity in bioassays against Rhammatocerus schistocercoides and showed unaltered lack of pathogenicity against larvae of the non-target insect Anticarsia gemmatalis. One co-transformant from four tested, however, showed a significant, but non-dose-dependent, elevation in virulence against Tenebrio molitor. PMID- 11024272 TI - Persistence of vancomycin-resistant enterococci (VRE) on Norwegian broiler farms. AB - Five Norwegian broiler farms previously identified as housing broilers carrying vancomycin-resistant enterococci (VRE) were examined for the presence of VRE 4 years after avoparcin was banned. Environmental samples were obtained from empty, cleaned broiler houses. Faecal samples were collected weekly from the flock housed after the environmental sampling. The hatchery from where the chicks originated was also sampled. VRE were found to be present in the farm environment after depopulation and cleanup of the broiler houses. Within 3 weeks after introduction to the farm, all broiler flocks tested positive for VRE. VRE were not isolated from the hatchery. PMID- 11024273 TI - Localization of the Leptospira interrogans metF gene on the CII secondary chromosome. AB - An open reading frame of 885 nucleotides was identified as the Leptospira interrogans metF gene. The deduced amino acid sequence (294 amino acids) showed similarities with Escherichia coli methylene tetrahydrofolate reductase (MetF or MTHFR) (33% identity) and with the N-terminal part of human MTHFR (33% identity). The L. interrogans metF gene complements an E. coli metF mutant to prototrophy, suggesting the functionality of the folate branch converging to form methionine. In addition, the L. interrogans MetF was found to be thermolabile. The metF gene belonged to the CII secondary chromosome, in contrast to the previously isolated metY and metX genes, which have been localized to the CI chromosome of Leptospira sp. PMID- 11024274 TI - An allelic series for the chalcone synthase locus in Arabidopsis. AB - Five new alleles of the Arabidopsis chalcone synthase (CHS) locus, tt4, have been characterized at the gene, protein, and end product levels as a genetic approach to understanding structure-function relationships in a key enzyme of plant secondary metabolism. Together with two previously described mutants, these tt4 lines represent one of the first allelic series for a central enzyme of the flavonoid pathway and include both null alleles and alleles with leaky, apparently temperature-sensitive, phenotypes. A variety of effects on accumulation of CHS protein and flavonoid glycosides were observed among these lines, including alterations in the apparent stability and activity of the enzyme. Assembly of the CHS homodimer also appeared to be impacted in several cases. A three-dimensional model of the Arabidopsis CHS protein, based on the recently determined structure for alfalfa CHS, predicts significant effects on protein structure or folding for several of the mutations. This allelic series should provide a useful genetic resource for ongoing studies of flavonoid enzyme structure, function, and subcellular organization. PMID- 11024275 TI - Genes and the physics of the DNA double-helix. AB - The processing of the genetic information stored in the double-helical DNA implies the separation of the two strands, the physics of which is described by the helix-coil transition model. Is there a relationship between genetic maps and DNA physical stability maps that plot the sequence-specific propensity for the thermal disruption of the double-helix? Here, with appropriate methodological formulations, such maps are derived for a large set of sequences, including complete genomes. The superposition of the two maps leads to a contrasted picture with correlations ranging between two extremes: from almost perfect (with the genes precisely delineated as stable regions) to more or less complete unrelatedness. The simplest explanation for the results is that the observed striking correlations correspond to the relics of a primeval organisation of the genetic message, with the physics of DNA playing a role in the delimitation of coding regions. In order to trace the evolutionary fate of this signal further, a detailed study of the yeast complete genome is performed. In this study, the superposition of the genetic and physical stability maps is examined in the light of information concerning gene duplication. On the basis of this analysis it is concluded that the 'signature' associated with the supposed archaic signal is in the process of being erased, most probably because the underlying feature is no longer under selective pressure. There are many evolutionary implications for the results presented and for their proposed interpretations, notably concerning models of mutational dynamics in relation to erasure processes. PMID- 11024276 TI - The physics of DNA and the annotation of the Plasmodium falciparum genome. AB - A gene identification procedure is formulated, based on large-scale structural analyses of genomic sequences. The structural property is the physical - thermal stability of the DNA double-helix, as described by the classical helix-coil model. The analyses are detailed for the Plasmodium falciparum genome, which represents one of the most difficult cases for the gene identification problem (notably because of the extreme AT-richness of the genome). In this genome, the coding domains (either uninterrupted genes or exons in split genes) are accurately identified as regions of high thermal stability. The conclusion is based on the study of the available cloned genes, of which 17 examples are described in detail. These examples demonstrate that the physical criterion is valid for the detection of coding regions whose lengths extend from a few base pairs up to several thousand base pairs. Accordingly, the structural analyses can provide a powerful and convenient tool for the identification of complex genes in the P. falciparum genome. The limits of such a scheme are discussed. The gene identification procedure is applied to the completely sequenced chromosomes (2 and 3), and the results are compared with the database annotations. The structural analyses suggest more or less extensive revision to the annotations, and also allow new putative genes to be identified in the chromosome sequences. Several examples of such new genes are described in detail. PMID- 11024277 TI - Precisely positioned nucleosomes are not essential for c-fos gene regulation in vivo. AB - Chromatin architecture plays a decisive role in many aspects of transcription regulation. We have tested the role of specific chromatin structures in c-fos gene regulation, using a gene transfer system based on episomes derived from the Epstein-Barr virus (EBV). This system reproduces in several respects the chromatin structure and regulation of the chromosomal c-fos gene. Using this approach, we first demonstrate that the pausing of RNA polymerase II downstream of the transcriptional start site does not require precisely positioned nucleosomes. Indeed, changing the pattern of MNase hypersensitive sites along the transcribed sequence does not perturb RNA polymerase II pausing or the regulation of the c-fos gene. Next, we show that a putative nucleosome positioned between the SIE/SRE elements (-300) and the CRE/TATA elements (-36) is not necessary for activation by a variety of inducers. Accordingly, total or partial deletion of the putative nucleosome sequence does not disturb c-fos regulation while the two regulatory sites flanking the nucleosome sequence remain hypersensitive to MNase. As described in this paper, EBV episomes are useful vectors to critically examine the role of the chromatin structure in gene transcription for human cells. PMID- 11024278 TI - RNA polymerase II promoter-proximal pausing upregulates c-fos gene expression. AB - Transcription elongation regulates c-fos expression in mouse and human cells. In the inactive state of the gene, RNA polymerases are engaged only in the promoter proximal region. Upon activation, RNA polymerases move efficiently along the complete gene. We have used Epstein-Barr virus (EBV) episomes as a gene transfer system to study the role of promoter-proximal pausing and transcript elongation in c-fos expression. We find that the sequence located immediately downstream of the transcriptional start site specifies pausing of RNA polymerases, dependent on both its orientation and position relative to the promoter. This sequence is, however, not necessary to maintain repression in the absence of a stimulus. As promoter-proximal pausing is therefore not a repression mechanism for the c-fos gene, the promoter and enhancer sequences are the main determinants of RNA polymerase elongation competence. Surprisingly, we find that promoter-proximal pausing further increases transcriptional levels from a variety of promoters. These observations lead us to hypothesize that promoter-proximal pausing of RNA polymerase II augments c-fos expression by allowing more efficient phosphorylation of the C-terminal domain of the large subunit. PMID- 11024279 TI - Double FYVE-containing protein 1 (DFCP1): isolation, cloning and characterization of a novel FYVE finger protein from a human bone marrow cDNA library. AB - Double FYVE-containing protein 1 (DFCP1) encodes a 777 amino acid protein that contains: (1) an N-terminal Cys-His cluster with some homology to many zinc finger domains; (2) a consensus sequence consistent with an ATP/GTP binding site; and (3) a C-terminal domain unique because it contains two zinc-binding FYVE domains. The gene, ZNFN2A1 (GenBank accession no. AF251025) was localized to chromosome 14q22-q24 and shown to be composed of 11 exons. Northern blot analysis revealed the presence of three different mRNA transcripts (4.2, 3 and 1.2kb). The two longer transcripts appear to be expressed in a variety of different tissues, especially in endocrine tissues, while the shorter messenger is limited to testis. Both of the larger transcripts are unusual due to the presence of a 463bp long 5' UTR. Furthermore, the 4.2kb transcript contains a non-standard polyadenylation consensus sequence while the 3kb transcript contains a standard consensus sequence but within the open reading frame. Following in vitro transfection of a DFCP1-containing expression construct, confocal microscopy studies showed a vesicular distribution of DFCP1 suggesting that this protein, like other FYVE-containing proteins, might be involved in membrane trafficking. PMID- 11024281 TI - Structure, expression, and chromosomal localization of the human gene encoding a germinal center-associated nuclear protein (GANP) that associates with MCM3 involved in the initiation of DNA replication. AB - A 210kDa protein named GANP is upregulated in germinal center (GC)-B cells in the spleen of antigen-immunized mouse. We studied a human ganp gene (hganp) encoding a putative polypeptide of 1980 amino acids. The carboxyl-terminal 721-amino-acid sequence of hGANP is identical to Map80, that is presumably generated by alternative splicing of hganp/Map80 gene. The genomic segment carrying hganp and Map80 genes was isolated, and the chromosomal location was determined on 21q22.3. Northern blot analysis with RNAs from various organs demonstrated a single band of 7kb hganp mRNA, which suggests a preferential transcription of hganp gene from the hganp/Map80 locus. The hGANP expression was upregulated in GCs of the tonsil, as demonstrated by in-situ RNA hybridization and immunohistochemical analyses. The hGANP, with the domain (Map-box) capable of binding to MCM3 in B cells, might be involved in regulation of cell-cycle progression and DNA replication of GC-B cells. PMID- 11024280 TI - Subtractive hybridization identifies chick-cripto, a novel EGF-CFC ortholog expressed during gastrulation, neurulation and early cardiogenesis. AB - EGF-CFC genes encode a novel class of extracellular, membrane-associated proteins that notably play an important role during vertebrate gastrulation. Whereas the two cysteine-rich domains that characterize these proteins, namely the extracellular EGF-like and the CFC domain, are known to be encoded by two evolutionarily conserved exons, it is generally assumed, based on weak primary sequence identity, that the remaining parts of the protein differ among vertebrates, suggesting that known members of the EGF-CFC family do not represent true orthologs. Here, by characterizing the full cDNA and genomic sequences of a new EGF-CFC gene in chick, and by comparing them with their counterparts in human (CRIPTO), mouse (cripto and cryptic), Xenopus (FRL-1) and zebrafish (one-eyed pinhead), we show that all EGF-CFC genes share an identical genomic organization over the entire coding region. Not only are the central two exons (coding for the EGF-like and CFC motifs) conserved, but also conserved are the total number of exons, their size, their intron phase and their correlation with discrete protein modules, in particular those modules that allow the EGF-CFC motif to become membrane-associated. Therefore, despite apparent divergence between their 5' and 3'-terminal exons, all known CRIPTO-related genes are structurally orthologous. We named this novel ortholog in bird, chick-cripto. We report the mRNA distribution of chick-cripto, which begins in the epiblast of the gastrula, with a pattern similar to EGF-CFC genes of other vertebrates. PMID- 11024282 TI - Isoforms of JSAP1 scaffold protein generated through alternative splicing. AB - We have identified four isoforms of c-Jun NH(2)-terminal kinase (JNK)/stress activated protein kinase-associated protein 1 (JSAP1), a scaffold protein that participates in JNK mitogen-activated protein kinase cascades, termed JSAP1a, JSAP1b, JSAP1c, and JSAP1d. The previously identified JSAP1 was renamed JSAP1a to avoid confusion. Analyses of the exon-intron structure of the jsap1 gene indicated that the isoforms are generated through alternative splicing involving exons 5 and 6. The mRNA expression levels of the JSAP1 isoforms differed among the mouse tissues examined. We also investigated the region of JSAP1 responsible for its interaction with JNK, and found that the JNK-binding domain is located between aa residues 201 and 217 in JSAP1a, which is encoded by part of exon 6. As all the JSAP1 isoforms contain this binding domain, we examined the binding affinity of the JSAP1 isoforms for JNK1, JNK2, and JNK3. JSAP1c and JSAP1d, which contain a 31-aa sequence not present in JSAP1a or JSAP1b, had a lower binding affinity for the JNKs, especially JNK3. These results suggest that JSAP1c and JSAP1d may attenuate the scaffolding activity of JSAP1a and/or JSAP1b in JNK cascades, especially the JNK3 cascades. PMID- 11024283 TI - Rev-binding aptamer and CMV promoter act as decoys to inhibit HIV replication. AB - We examined whether the antiviral effect of an HIV-1 Rev-binding aptamer [RBE(apt)] could be enhanced by a ribozyme directed against the HIV-1 env gene, and whether the antiviral activity was affected by different promoters. The efficacy of the aptamer and ribozyme DNAs was tested in HeLa cells co-transfected with the HIV-1 proviral clones, HXBDeltaBgl or pNL4-3, using transferrin lipoplexes. The RBE(apt) and anti-env ribozyme genes were inserted into the pTZU6+27 plasmid, or constructed under the control of the human cytomegalovirus (CMV) or Rous sarcoma virus (RSV) promoters. The parental vector plasmids were used as controls. Co-transfection of the pTZU6+27 RBE(apt) plasmid with HXBDeltaBgl, or pNL4-3, at a weight ratio of 5:1, inhibited p24 production by 70 and 45%, respectively. The RSV RBE(apt) plasmid co-transfected with either HIV clone, at the same weight ratio, reduced viral production by 88%. The addition of the anti-env ribozyme to the RSV RBE(apt) did not enhance its antiviral activity. When the constructs were under the control of the CMV promoter, the expression of the HIV plasmids was very low and was independent of the presence of the RBE(apt). Thus, the effect of the RBE(apt) was strongly dependent on the promoter of the tested construct. The anti-HIV activity of the CMV RBE(apt) construct was non-specific, because co-transfection with either pCMV. SPORT-betagal or pCMVlacZ significantly suppressed HIV production from the HIV proviral clones. The reduction in p24 could not be attributed to the non-specific toxicity of the transfection procedure. Transfection of acutely HIV-infected HeLa-CD4 cells with pCMV.SPORT-betagal reduced the p24 level by 35%, while the expression of the U6 RBE(apt) did not affect p24 production. The suppression of HIV production from the HIV proviral clones by the CMV promoter constructs in the co-transfection assays may be explained by competition for transcription factors (TFs) between HIV and CMV promoters. This observation points to the potential for misleading results in co-transfections involving CMV constructs and HIV. PMID- 11024284 TI - Isolation and characterization of disease resistance gene homologues from rice cultivar IR64. AB - We initiated a search for disease resistance (R) gene homologues in rice cultivar IR64, one of the most agronomically important rice varieties in the world, with the assumption that some of these homologues would correspond to previously identified disease resistance loci. A family of rice R gene homologues was identified using the Arabidopsis NBS-LRR disease resistance gene RPS2 as a hybridization probe. Because member genes of this rice R gene family exhibit features characteristic of the NBS-LRR class of resistance genes, the family was given the name NRH (for NBS-LRR resistance gene homologues). Three members of the NRH family, NRH1, NRH2, and NRH3, were cloned and studied in detail. In IR64, NRH1 and NRH2 appear to encode full-length polypeptides, whereas NRH3 is prematurely truncated with a stop codon generated by a frameshift. NRH1 maps on chromosome 5, and NRH2 and NRH3 are less than 48kb apart on chromosome 11. Although NRH1, NRH2, and NRH3 map to regions of the rice genome where disease resistance loci to Xanthomonas oryzae pv. oryzae (Xoo) have been identified, susceptible rice varieties transformed with either NRH1 or NRH2 failed to exhibit increased resistance to a set of well-characterized Xoo strains. PMID- 11024285 TI - Intron-exon structure and gene copy number of a gene encoding for a membrane intrinsic light-harvesting polypeptide of the red alga Galdieria sulphuraria. AB - Genes for light-harvesting proteins (lhc genes) of higher plants are well examined. However, little is known about the corresponding genes of algae, although this knowledge might give valuable information about the evolution of photosynthetic antennae. In the case of rhodophytes only two cDNA sequences from a single organism, Porphyridium cruentum, have been published. Here we describe an additional sequence from another species, the thermo-acidophilic red alga Galdieria sulphuraria. For the first time also a genomic sequence for a red algal lhc gene is presented. From a cDNA library of G. sulphuraria we isolated a clone containing an open reading frame for a protein of 302 amino acids with a deduced molecular mass of 33.86kDa. It shares major structural features with eukaryotic light-harvesting polypeptides. A proposed cleavage site between transit peptide and mature protein gives rise to a transit peptide of 119 amino acids and a mature protein of 183 residues. Hydropathy analysis suggests that the mature protein consists of three transmembrane helices. Several amino acid residues supposed to bind chlorophyll a and chlorophyll b in higher plants are conserved. The protein shows up to 69% identity and 81% similarity to the Porphyridium polypeptides in the transmembrane helices 1 and 3. Using oligonucleotides annealing in the regions of the start and stop codons of the gene as primers, a DNA sequence was amplified from nuclear G. sulphuraria DNA by PCR. Compared with the cDNA clone, this sequence contains five additional intervening DNA strings of 50-74bp length. Four of them show typical features of spliceosomal introns with GT-AG borders, and the fifth differs by starting with GC. Three of the supposed introns are located in similar positions as introns of higher plant light harvesting proteins. Southern blotting and hybridization experiments indicate that G. sulphuraria contains at least three copies of this gene. PMID- 11024286 TI - Organization of the human gene encoding heterogeneous nuclear ribonucleoprotein type I (hnRNP I) and characterization of hnRNP I related pseudogene. AB - The human gene hnRNPI encoding the heterogeneous nuclear ribonucleoprotein type I, an alternative splicing modulator of tissue-specific transcripts, also known as PTB (polypyrimidine tract-binding protein), was recently mapped on chromosome 14, as well as on chromosome 19, suggesting that two closely related copies of the same gene might exist in the human genome. We report here that the gene localized on chromosome 14 corresponds to a highly homologous processed pseudogene related to hnRNPI gene (psihnRNPI). Analysis by RT-PCR and by EST database comparison indicates that psihnRNPI is not expressed. In this report we have also analyzed the organization of the actual hnRNPI gene localized on chromosome 19. The DNA sequence at the intron-exon boundaries unveiled the possible mechanism by which three isoforms of the protein (namely hnRNPI, PTB2 and PTB3) are generated by means of alternative splicing of the same hnRNPI gene transcript. PMID- 11024287 TI - Identification of mRNAs expressed in tumor-infiltrating lymphocytes by a strategy for rapid and high throughput screening. AB - Most gene expression methods often involve cumbersome steps or use expensive facilities. Additionally, some of the techniques, such as cDNA biochip, cannot define the sub-population of tissue from which the amplified cDNA was made. Here we present a rapid and high throughput screening method for analyzing the pattern of gene expression of tumor-infiltrating lymphocyte (TIL), which can minimize manipulations in cloned DNA sequencing and in bioinformatics. The pattern of TIL gene expression was studied in one ovarian cancer and one liver cancer. Our results have demonstrated that TILs have three different gene expression profiles: the first set of genes is involved in cell proliferation and mitogenic stimulation, such as c-myc and IL-8, LD78, MIP-1beta, insulin-induced protein and AH-receptor; the second set of genes includes those involved in attachment of lymphocytes to endothelium and extravasation into tumor tissues such as P selectin ligand and integrin; and the third set, which includes genes such as the perforin, FAS ligand and granzyme B, is related to cytotoxic function to tumor cells. The patterns of TIL gene expression obtained from two specimens are marginally different and can be used in explaining the basis of molecular mechanisms regulating cellular interactions and cytotoxicity. PMID- 11024288 TI - Analysis of the 2,4-dichlorophenoxyacetic acid-degradative plasmid pEST4011 of Achromobacter xylosoxidans subsp. denitrificans strain EST4002. AB - The 2,4-dichlorophenoxyacetic acid (2,4-D)-degradative bacterium Achromobacter xylosoxidans subsp. denitrificans strain EST4002, isolated in Estonia more than 10years ago, was found to contain the 70kb plasmid pEST4011 that is responsible for the bacterium having had obtained a stable 2,4-D(+) phenotype. The tfd-like genes for 2, 4-D degradation of the strain EST4002 were located on a 10.5kb region of pEST4011, but without functional genes coding for chloromuconate cycloisomerase and chlorodienelactone hydrolase. The latter two genes are probably encoded by homologous, tcb-like genes, located elsewhere on pEST4011. We also present evidence of two copies of insertion element IS1071-like sequences on pEST4011. IS1071 is a class II (Tn3 family) insertion element, associated with different catabolic genes and operons and globally distributed in the recent past. We speculate that this insertion element might have had a role in the formation of plasmid pEST4011. The 28kb plasmid pEST4012 is generated by deletion from pEST4011 when cells of A. xylosoxidans EST4002 are grown in the absence of 2,4-D in growth medium. We propose that this is the result of homologous recombination between the two putative copies of IS1071-like sequences on pEST4011. PMID- 11024289 TI - Molecular characterization of a new human T-box gene (TBX22) located in xq21.1 encoding a protein containing a truncated T-domain. AB - We are conducting a systematic transcriptional mapping of the Xq12-q21 region of the human X chromosome in order to identify new genes potentially involved in X linked mental retardation phenotypes. In silico analysis using the sequence of the genomic clones originating from this region of the human X chromosome allowed us to characterize a new gene belonging to the T-box family of transcriptional regulators. These genes were shown to be critical for proper development of both vertebrates and invertebrates. We show here that this new gene, called TBX22, is composed of seven exons spanning 8.7 kilobases of genomic DNA in Xq21.1. The TBX22 mRNA is 2099 base pairs long and encodes a 400-amino-acids protein containing a T-domain in its NH(2)-terminal region which has the unique feature of missing 20 amino-acids relative to the other known T-domains. TBX22 transcripts were exclusively found in a human fetal cDNA library and no homologous gene could be detected in the mouse genome. In addition, phylogenetic studies performed using all the known T-domain-containing proteins show that TBX22 is not directly related to any of them. These data indicate that TBX22 may be the first identified member of a new family of T-domain-containing proteins. PMID- 11024290 TI - Regulated gene expression in Staphylococcus aureus for identifying conditional lethal phenotypes and antibiotic mode of action. AB - Selectively regulating gene expression in bacteria has provided an important tool for studying gene function. However, well-regulated gene control systems have been restricted primarily for use in laboratory non-pathogenic strains of bacteria (e.g. Escherichia coli, Bacillus subtilis). The development of analogous systems for use in bacterial pathogens such as Staphylococcus aureus would significantly enhance our ability to examine the contribution of any given gene product to pathogen growth and viability. In this report, we adapt, examine and compare three regulated gene expression systems in S. aureus, which had previously been used in B. subtilis. We demonstrate that all three systems function and exhibit titratable induction, together covering a dynamic range of gene expression of approximately 3000-fold. This dynamic range correlates well with the physiological expression levels of cellular proteins. Importantly, we show that one of these systems, the Spac system, is particularly useful for examining gene essentiality and creating specific conditional lethal phenotypes. Moreover, we find that titration of selective target gene products using this system allows direct demonstration of antibiotic mode of action. PMID- 11024291 TI - Cloning and expression of type II collagen mRNA: evaluation as a candidate for canine oculo-skeletal dysplasia. AB - The disease phenotype of oculo-skeletal dysplasia (OSD) detected in Labrador retrievers and Samoyeds shows a large degree of similarity with human Stickler and Kniest dysplasia. Type II collagen (COL2A1) mRNA, which is defective in a larger number of Stickler and Kniest patients, has been cloned and characterized from normal dog. The amino acid sequence of the canine type II procollagen is predicted to contain 1487 residues, with high degree of homology with its human homologue, and maintains all the characteristic structural domains. In addition to cartilage, expression of COL2A1 has also been detected in canine retina and testes. In testes, the N-propeptide region of COL2A1 displayed differential splicing and expressed both splice variants, IIA (with exon 2) and IIB (without exon 2), suggesting the importance of both forms in testis maturation and maintenance. Despite a severe decrease of type II collagen protein in the vitreous of OSD affected Labrador retrievers, COL2A1 gene has been excluded from having any causal association with the disease locus by linkage analysis. Using an intragenic RFLP marker, COL2A1 gene has also been tested as a candidate gene for the non-allelic form of the other canine OSD identified in Samoyeds, and excluded by linkage analysis. Oculo-skeletal dysplastic Labrador retriever and Samoyed provide two animal models for chondrodysplasia with genetic heterogeneity. PMID- 11024292 TI - Tunicates have unusual nuclear lamins with a large deletion in the carboxyterminal tail domain. AB - Lamins are essential proteins of metazoa. They give rise to the nuclear lamina lining the nucleoplasmic face of the inner nuclear membrane. Here we report the isolation of complete lamin cDNA clones from three urochordate (tunicate) libraries - adult Ciona intestinalis, the tailbud stage of Styela clava and the gastrula stage of Molgula oculata. Lamins L1 and L2 of adult Ciona are derived from two distinct genes. The sequence of the 3' part of the Ciona lamin L1 gene shows that the alpha and beta variants of lamin L1 in Ciona and Styela arise by alternative choice of the 5' splice site at the last intron. Strikingly, all urochordate sequences reveal a 90 residue deletion which removes nearly the entire 105-box. This region is the only long sequence homology segment in the carboxyterminal tail domain of lamins from animals as diverse as Hydra, Drosophila, Priapulus, Caenorhabditis elegans, several echinoderms, the cephalochordate Branchiostoma and various vertebrates. We discuss this unexpected plasticity of lamin sequences as a urochordate specific marker. To increase the database for the chordates we completed the partial sequence of the Branchiostoma lamin by the N-terminal head and central rod domains. The molecular phylogenetic analysis of the metazoan lamin sequences emphasises the monophyletic nature of the chordates in line with the morphological evidence. PMID- 11024293 TI - Analysis of the genes flanking xabB: a methyltransferase gene is involved in albicidin biosynthesis in Xanthomonas albilineans. AB - Transposon mutagenesis and complementation studies previously identified a gene (xabB) for a large (526kDa) polyketide-peptide synthase required for biosynthesis of albicidin antibiotics and phytotoxins in the sugarcane leaf scald pathogen Xanthomonas albilineans. A cistron immediately downstream from xabB encodes a polypeptide of 343aa containing three conserved motifs characteristic of a family of S-adenosyl-L-methionine (SAM)-dependent O-methyltransferases. Insertional mutagenesis and complementation indicate that the product of this cistron (designated xabC) is essential for albicidin production, and that there is no other required downstream cistron. The xabB promoter region is bidirectional, and insertional mutagenesis of the first open reading frame (ORF) in the divergent gene also blocks albicidin biosynthesis. This divergent ORF (designated thp) encodes a protein of 239aa displaying high similarity to several IS21-like transposition helper proteins. The thp cistron is not located in a recognizable transposon, and is probably a remnant from a past transposition event that may have contributed to the development of the albicidin biosynthetic gene cluster. Failure of 'in trans' complementation of thp indicates that a downstream cistron transcribed with thp is required for albicidin biosynthesis. PMID- 11024294 TI - Progressive inactivation of the haploid expressed gene for the sperm-specific endozepine-like peptide (ELP) through primate evolution. AB - The endozepine-like peptide (ELP) is a novel intracellular molecule which is expressed in high amounts at both mRNA and protein levels very specifically in late haploid male germ cells. It is closely related to the ubiquitous acyl-CoA binding protein, is highly conserved, shares a similar ability to bind mid-long chain acyl-CoA, and is thus likely to be involved in mature sperm metabolism. While it has been characterized from diverse mammals, it has so far not been possible to identify an equivalent molecule in the primate testis. Using a PCR approach, combined with cDNA cloning and Northern hybridization, testicular transcripts and/or genomic DNA were analysed for different primate species, including human. In the marmoset and cynomolgus macaque normally structured transcripts appear to be expressed, though at a low level. In the human testis, two rare transcripts were characterized, hELP1 and hELP2, the products of independent duplicated genes. Both transcripts were longer than in non-mammalian species, included frame-shift mutations and substantial sequence insertions, preventing the translation of a sensible protein. Genomic PCR analysis of three anthropoid species, chimpanzee, gorilla and orangutan, showed the presence of a similarly mutated hELP1 gene. Only in the gorilla was a hELP2 gene identified, apparently lacking the frame-shift mutation, and thus potentially able to give rise to a functional ELP protein. Taken together, these results show that during primate evolution there has been a progressive inactivation of the ELP gene, initially with a down-regulation in lower primates, and subsequently with inactivating mutations in the open reading frame. At some time during simian evolution prior to these mutations there has been a gene duplication, though this second gene has also become inactivated in humans. In its pattern of evolution the ELP gene shows similarities with the MDC/fertilin family, whose members are also considered essential components of haploid sperm in non-primates, but which are progressively inactivated in anthropoids and humans. We should like to speculate that the established subfertility of the human male may not be a recent event, but the consequence of a longer evolutionary process whereby primates have traded off absolute fertility against social or sexual advantages. PMID- 11024295 TI - Human nectin3/PRR3: a novel member of the PVR/PRR/nectin family that interacts with afadin. AB - We have isolated nectin3/PRR3, the fourth human member of the nectin/PRR family, also described as the alpha herpes virus receptor family. Nectin/PRR members are adhesion molecules expressed at intercellular junctions. Nectin3/PRR3 is a transmembrane protein, whose extracellular region contains three Ig-like domains (V, C and C) and shares approximately 30% identity with the other members. It is mainly expressed in testis and placental tissues. SDS-PAGE analyses demonstrate that nectin3/PRR3 has a molecular weight of 83kDa. Nectin1/PRR1L and nectin2/PRR2S and L were found to be specifically expressed at the intercellular junctions. This localization is in part due to the interaction of the C-terminal part of these receptors (ended by the consensus sequence A/EXYV) and the PDZ domain of afadin. In this report we demonstrate that the nectin3/PRR3 receptor carries the A/EXYV consensus sequence and interacts in vivo with both long and short isoforms of afadin. These results suggest that the human nectin3/PRR3 is a new afadin-associated molecule. PMID- 11024297 TI - X-chromosome inactivation: lessons from transgenic mice. AB - X-chromosome inactivation (XCI) is the process by which mammals perform dosage compensation of X-linked gene products between XY males and XX females, resulting in the transcriptional silencing of all but one X chromosome per diploid cell. XCI involves counting the X chromosomes in a cell, randomly choosing those to be inactivated, spreading the inactivation signal in cis throughout the chromosome, and maintaining the inactive state of those X chromosomes during cell divisions thereafter. How the cell performs all these tasks is a fascinating problem and, together with epigenetic inheritance, a basic cellular mechanism that remains to be fully understood. In this review, we describe recent experiments aimed at understanding the first events of XCI and propose a model for initiation of XCI. PMID- 11024296 TI - Structure, chromosomal localization, and expression of the Drosophila melanogaster gene encoding sepiapterin reductase. AB - We have isolated and characterized a Drosophila melanogaster gene encoding the sepiapterin reductase (SR). The gene does not have introns. The 5'- and 3'-RACE analysis, which determined the transcription start point (tsp) and polyadenylation site, respectively, showed that the gene produces single mRNA species. The potential promoter region lacks distinct TATAAA or CCAAT box consensus sequences. RNA blot analysis revealed that the gene encodes a 1.4kb transcript that could be detected throughout development and in both heads and bodies of adults. The Drosophila SR gene maps to 15A on the X chromosome. PMID- 11024298 TI - A human and a mouse anti-idiotypic antibody specific for human T14(+) anti-DNA antibodies reconstructed by phage display. AB - Little is known about human anti-idiotypic antibodies. Phage display methodology was used to reconstruct these antibodies from lupus patients, which recognize a subset (T14(+)) of anti-DNA antibodies. Antigen-specific B cells were isolated from the blood using a peptide based on a complementarity determining region (V(H)CDR3) of the prototypic T14(+) antibody. cDNA fragments of the V(H) and V(L) genes prepared from the cells were expressed as phage displayed single chain Fv (scFv) fragments using the pCANTAB-5E phagemid vector. From a reactive clone obtained, the Ig genes used were identified to be V(H)3, D5-D3, J(H)4b, V(kappa)I and J(kappa)2. The heavy chain was highly mutated, especially in CDR3, which bears mutations mostly of the replacement type; this region is also unusual in being extremely long due to a D-D fusion. In contrast, a mouse hybridoma antibody, made to the same T14(+) peptide and transformed as a scFv fragment, uses a short V(H)CDR3 comprising five amino acids, three of which are tyrosines. Tyrosines may be important for antigen binding because two of these also exist in the human V(H)CDR3. The light chains of both antibodies may also contribute to the specificity of the protein, because their V(L) segments, including the CDRs, are highly homologous to each other. PMID- 11024299 TI - Sequence and transcription patterns of 60S ribosomal protein P0, a diapause regulated AP endonuclease in the flesh fly, Sarcophaga crassipalpis. AB - We have isolated and sequenced a 1308bp clone from a pupal brain cDNA library of the flesh fly, Sarcophaga crassipalpis, showing 97% amino acid (aa) sequence similarity to Ceratitis capitata 60S acidic ribosomal protein P0 (CcP0) and 93% aa sequence similiarity to Drosophila melanogaster P0 (DmP0). DmP0 is a multifunctional protein necessary for efficient protein translation of the 60S ribosome as well as DNA repair via AP3 endonuclease activity. In this study, we observed that S. crassipalpis P0 (ScP0) is cyclically regulated throughout the fly's overwintering pupal diapause. Expression of ScP0 cycles out of phase with the 4day cycles of O(2) consumption: the peak day of O(2) consumption is characterized by low ScP0 expression, while high expression is noted during the trough of the O(2) consumption cycle. The O(2) cycles, which are in turn driven by cycles of juvenile hormone (JH), can be eliminated by application of a JH analog (JHA). Pupae rendered acyclic with a JHA application consume O(2) at a constant high rate and ScP0 is consistently downregulated. Our findings thus suggest that the cyclic nature of ScP0 regulation during pupal diapause is linked to the JH-mediated metabolic cycles characteristic of this species. PMID- 11024300 TI - A novel glutamine-rich putative transcriptional adaptor protein (TIG-1), preferentially expressed in placental and bone-marrow tissues. AB - We used a subtractive hybridization protocol to identify novel expressed sequence tags (ESTs) corresponding to mRNAs whose expression was induced upon exposure of the human leukemia cell line K562 to the phorbol ester 12-O-tetradecanolyphorbol 13-acetate (TPA). The complete open reading frame of one of the novel ESTs, named TIG-1, was obtained by screening K562 cell and placental cDNA libraries. The deduced open reading frame of the TIG-1 cDNA encodes for a glutamine repeat-rich protein with a predicted molecular weight of 63kDa. The predicted open reading frame also contains a consensus bipartite nuclear localization signal, though no specific DNA-binding domain is found. The corresponding TIG-1 mRNA is ubiquitously expressed. Placental tissue expresses the TIG-1 mRNA 200 times more than the lowest expressing tissues such as kidney and lung. There is also preferential TIG-1 mRNA expression in cells of bone-marrow lineage.In-vitro transcription/translation of the TIG-1 cDNA yielded a polypeptide with an apparent molecular weight of 97kDa. Using polyclonal antibodies obtained from a rabbit immunized with the carboxy-terminal portion of bacterially expressed TIG-1 protein, a polypeptide with molecular weight of 97kDa was identified by Western blot analyses of protein lysates obtained from K562 cells. Cotransfection assays of K562 cells, using a GAL4-TIG-1 fusion gene and GAL4 operator-CAT, indicate that the TIG-1 protein may have transcriptional regulatory activity when tethered to DNA. We hypothesize that this novel glutamine-rich protein participates in a protein complex that regulates gene transcription. It has been demonstrated by Naar et al. (Naar, A.M., Beaurang, P.A., Zhou, S., Abraham, S., Solomon, W.B., Tjian, R., 1999, Composite co-activator ARC mediates chromatin-directed transcriptional activation. Nature 398, 828-830) that the amino acid sequences of peptide fragments obtained from a polypeptide found in a complex of proteins that alters chromatin structure (ARC) are identical to portions of the deduced open reading frame of TIG-1 mRNA. PMID- 11024301 TI - Cloning of telomere-associated DNA using single-specific-primer polymerase chain reaction provides evidence for a conserved sequence directly adjacent to Theileria parva telomeric repeats. AB - Telomere-associated (TA) DNA sequences of the intracellular protozoan parasite Theileria parva were isolated by a novel strategy using a modified version of single-specific-primer polymerase chain reaction (SSP-PCR). Nucleotide sequences of non-coding TA DNA from three telomeres (6017bp, 2435bp and 4859bp) contained no extensive tracts of repetitive DNA. Long open reading frames (ORFs) were present at the centromeric ends of two of the TA sequences, the 3' ends of the closest ORFs being only 2670bp and 2719bp from the telomeric repeats. There were regions of significant similarity between the nucleotide sequences of the non coding regions of different telomeres. The longest region of similarity was a virtually identical 1650bp domain, located directly adjacent to the telomeric repeats of two separate telomeres. Comparison of the telomere proximal sequences defined in this study and two additional T. parva telomeres, whose sequences were determined previously, resulted in identification of a single copy 141bp conserved sequence directly adjacent to the telomeric repeats. The conserved sequence is present at all five T. parva telomeres that have been characterised. The only organism currently known to have a single copy conserved sequence located adjacent to the telomeric repeats is another intracellular protozoan, Leishmania braziliensis. PMID- 11024302 TI - Multiple variants of the peroxisome proliferator-activated receptor (PPAR) gamma are expressed in the liver of atlantic salmon (Salmo salar). AB - A full-length cDNA encoding the peroxisome proliferator-activated receptor (PPAR) has for the first time been characterized from a fish species. The Atlantic salmon PPARgamma cDNA of 2528 nucleotides (nt) was amplified from liver mRNA by reverse transcription (RT)-polymerase chain reaction (PCR). The deduced protein of 544 amino acids (aa) shares approximately 47% overall sequence identity with mammalian PPARgamma. The N-terminal A/B region contains a repeated decapeptide motif and shows a low homology with other PPARs. In contrast, the central DNA binding domain (DBD) and the C-terminal ligand-binding domain (LBD) show a high sequence identity to mammalian and Xenopus PPARgamma. The salmon PPARgamma LBD contains nine additional residues in a flexible loop that might affect ligand binding. Northern blot analysis of salmon liver RNA revealed a prominent transcript of about 1.7 kilo bases (kb), in addition to several mRNA species of about 2.4-2.6kb, which is consistent with the presence of multiple putative polyadenylation sites in the 3' untranslated region (UTR) of the 2528nt long PPARgamma cDNA. Two additional PPARgamma cDNAs of 1719 and 2357nt were then isolated. The 2357nt long transcript encodes full-length PPARgamma and seems to be ubiquitously expressed in salmon, whereas the liver-specific transcript of 1719nt encodes a truncated variant of PPARgamma. The truncated form lacks 39 C terminal residues including the conserved activation function-2 (AF-2) motif, known to be associated with crucial cofactors. Three-dimensional modelling studies indicated that the C-terminal truncation would result in important alterations of the ligand-binding pocket. The presence of a truncated form with drastic changes in both ligand- and cofactor-binding sites is likely to modulate PPARgamma activity in salmon liver. PMID- 11024303 TI - Purple acid phosphatases from bacteria: similarities to mammalian and plant enzymes. AB - Mammalian and plant purple acid phosphatases have similar active site structures despite low sequence identity (<20%). Although no bacterial enzyme has been purified, a sequence database search revealed that genes that could encode potential purple acid phosphatases may be restricted to a small number of organisms (i.e. myco- and cyanobacteria). Analysis of their deduced amino acid sequences and predicted secondary structures indicates that the cyanobacterial enzyme is similar to both the mammalian and the recently discovered low-molecular weight plant purple acid phosphatases, while the mycobacterial enzyme is homologous to the fungal and high-molecular-weight plant purple acid phosphatases. Homology models indicate that both bacterial proteins appear to be similar to mammalian purple acid phosphatases in the immediate vicinity of the active site. It is likely that these enzymes act as Fenton-type catalysts in order to prevent damage caused by reactive oxygen species generated by invaded host cells (M. tuberculosis) or by the light-harvesting complex (Synechocystis sp.). PMID- 11024304 TI - HIV disease in the year 2000. PMID- 11024305 TI - Global epidemiology of HIV infection and AIDS. PMID- 11024307 TI - Immunopathogenesis of HIV-1 infection. PMID- 11024306 TI - Natural history of HIV-1 infection. PMID- 11024308 TI - HIV disease: mucocutaneous fungal infections in HIV disease. PMID- 11024309 TI - Cutaneous viral infections in HIV-infected individuals. PMID- 11024310 TI - AIDS and the skin: parasitic diseases. PMID- 11024311 TI - Systemic manifestations of HIV infection. PMID- 11024312 TI - Oral manifestations of HIV disease. PMID- 11024314 TI - New developments in the etiopathogenesis and treatment of HIV-related Kaposi's sarcoma. PMID- 11024313 TI - Manifestations of HIV disease in children. PMID- 11024315 TI - Pruritus and cutaneous inflammatory conditions in HIV disease. PMID- 11024316 TI - Adverse cutaneous drug reactions in patients with human immunodeficiency virus-1 infection. PMID- 11024317 TI - Color atlas: aids and the skin PMID- 11024318 TI - Immunization in the new millennium: meeting the challenge to realize the promise. PMID- 11024319 TI - Calling the shots: immunization finance policies and practices. Executive summary of the report of the Institute of Medicine. AB - Federal, state, and private-sector investments in vaccine purchases and immunization programs are lagging behind emerging opportunities to reduce the risks of vaccine-preventable disease. Although federal assistance to the states for immunization programs and data collection efforts rapidly expanded in the early part of the 1990s, significant cutbacks have occurred in the last 5 years that have reduced the size of state grant awards by more than 50% from their highest point. During this same period, the vaccine delivery system for children and adults has become more complex and fragmented. This combination of new challenges and reduced resources has led to instability in the public health infrastructure that supports the U. S. immunization system. Many states have reduced the scale of their immunization programs and currently lack adequate strength in areas such as data collection among at-risk populations, strategic planning, program coordination, and assessment of immunization status in communities that are served by multiple health care providers. If unmet immunization needs are not identified and addressed, states will have difficulty in achieving the national goal of 90% coverage by the year 2010 for completion of the childhood immunization series for young children. Furthermore, state and national coverage rates, which reached record levels for vaccines in widespread use (79%, 1998), can be expected to decline and preventable disease outbreaks may occur as a result, particularly among persons who are vulnerable to vaccine preventable disease because of their underimmunization status. The Institute of Medicine (IOM) Committee on Immunization Finance Policies and Practices has therefore concluded that a renewal and strengthening of the federal and state immunization partnership is necessary. The goal of this renewed partnership is to prevent infectious disease; to monitor, sustain, and improve vaccine coverage rates for child and adult populations within more numerous and increasingly diversified health care settings; and to respond to vaccine-safety concerns. To achieve this renewal, states require a consistent strategy, additional funds, and a multiyear finance plan that can help expedite the delivery of new vaccines; strengthen the immunization assessment, assurance, and policy development functions in each state; and adapt childhood immunization programs to serve the needs of new age groups (especially adults with chronic diseases) in different health care environments. The IOM committee recommends that federal and state governments adopt a national finance strategy that would allocate $1.5 billion in federal and state resources over the first 5 years to strengthen the infrastructure for child and adult immunization-an annual increase of $175 million over current spending levels. These resources would consist of $200 million per year in state infrastructure grants awarded by the Centers for Disease Control and Prevention (the Section 317 program) and an additional $100 million per year in increased state contributions. The committee also recommends that the Congress replace the current discretionary Section 317 grants with a formula approach for state immunization grant awards to improve the targeting and stability of federal immunization grants. The formula should provide a base level of support to all states, as well as additional amounts related to each state's need, capacity, and performance. The committee further recommends that Congress introduce a state match requirement for the receipt of increased federal funds to help strengthen and stabilize the infrastructure that supports long-term public health assessment, assurance, and policy development efforts. (ABSTRACT TRUNCATED) PMID- 11024320 TI - Successful control of vaccine-preventable diseases requires more than vaccines. PMID- 11024321 TI - If we can map the human genome, we can immunize Chicago's children. PMID- 11024322 TI - The view from state and local programs. PMID- 11024323 TI - The public-private dynamics of national immunization policy. PMID- 11024324 TI - Vaccine coverage: access and administration. PMID- 11024325 TI - Vaccine financing: the view of managed care. PMID- 11024326 TI - The best and worst of times: use of adult immunizations. PMID- 11024327 TI - Improving immunization surveillance and performance monitoring. PMID- 11024328 TI - Making immunization programs work for children and pediatricians in North Carolina. PMID- 11024329 TI - State-level perspectives on immunization policies, practices, and program financing in the 1990s. AB - This article reports on a series of structured interviews with immunization program officials in all 50 states regarding the effects of changes in federal policies and funding in the 1990s on the goals, priorities, and activities of state immunization programs. The purchase of vaccines is a major component of all state immunization programs. The Vaccines for Children (VFC) program, implemented in 1994, has become the primary source of vaccine purchase support in almost all states. A concern of many state immunization programs is their ability to ensure that vaccines are available to children who are not VFC eligible.State immunization programs also are involved in a myriad of activities necessary to ensure that children are adequately and appropriately immunized (e.g. , vaccine administration, outreach to parents). Federal funding to support these activities increased significantly during the mid-1990s, but was substantially reduced beginning in 1997. Because of these funding decreases, most states had to reduce the scale and scope of their immunization activities.State-level funding support for immunization programs varies, with state governments more likely to support vaccine purchase than immunization activities. Immunization will never be completed. Along with each new birth cohort, changes to the primary immunization schedule (i.e., addition of new vaccines and expansion of existing recommendations to encompass broader target groups) create ongoing needs for vaccine purchase and other immunization activities. Long-term immunization planning must reflect these continually expanding needs. PMID- 11024330 TI - Federalism, entitlements, and discretionary grants: the fiscal context of national support for immunization programs. AB - This paper defines the appropriation and resource allocation structure of federal government programs that deliver vaccines to the public through state and local governments, with a special focus on the Centers for Disease Control and Prevention (CDC) Section 317 program. The paper places these programs in the context of the overall intergovernmental finance system of the United States, and the manner in which that system has responded to societal changes in other areas, such as unemployment insurance and environmental protection. The paper concludes that a changing environment for vaccine financing, and uncertain appropriations for state government immunization infrastructure, may have made the current CDC Section 317 program inadequate. Increased entitlement resources for federally funded vaccines have not been matched with increased and stable resources for the balance of program costs. To the contrary, funding provided to the Section 317 program has proven unstable, and its discretionary appropriations have proven vulnerable to both executive and legislative branch earmarking for such activities as overseas disease eradication or home state earmarking by congressional appropriations subcommittee members. In addition, the rigidity of some CDC program requirements makes it difficult for states to effectively use program funds. Consideration should be given to strengthening the Section 317 program and its relationships to other federal aid programs to ensure that adequate immunization protections are provided in all states to adequately protect all citizens. PMID- 11024331 TI - Findings from case studies of state and local immunization programs. AB - BACKGROUND: As part of its examination of federal support for immunization services during the past decade, the Institute of Medicine (IOM) Committee on Immunization Finance Policies and Practices (IFPP) commissioned eight case studies of the states of Alabama, Maine, Michigan, New Jersey, North Carolina, Texas, and Washington; and a two-county study of Los Angeles and San Diego in California. Specifically, the IOM Committee and these studies reviewed the use of Section 317 grants by the states. Section 317 is a discretionary grant program that supports vaccine purchase and other immunization-related program activities. These studies afforded the Committee an in-depth look at local policy choices, the performance of immunization programs, and federal and state spending for immunization during the past decade. METHODS: The case-study reports were developed through interviews with state and local health department officials, including immunization program directors, Medicaid agency staff, budget analysts, and Centers for Disease Control and Prevention public health advisors to the jurisdiction. Other sources included state and federal administrative records and secondary sources on background factors and state-level trends. The case studies were supplemented by site visits to Detroit, Houston, Los Angeles, Newark, and San Diego. OBSERVATIONS: The nature of immunization "infrastructure" supported by the Section 317 program is shifting from primarily service delivery to a broader set of roles that puts the public effort at the head of a broad immunization partnership among public health, health financing, and other entities in both the public and private sectors. The rate and intensity of transition vary across the case-study areas. In the emerging pattern, service delivery increasingly takes place in the private sector and is related to managed care. "Infrastructure" is moving beyond supporting a core state staff and local health department service delivery to include such activities as immunization registries, quality improvement, and coordination with programs outside public health agencies. At the same time, the recent decline in federal Section 317 support is forcing difficult choices between old and new activities at the state and local levels. CONCLUSIONS: Immunization programs function as an organic component of the local health care financing and delivery systems of which they are a part. Immunization efforts are organized and conducted within distinctive state and local fiscal, economic, and health care contexts. Section 317 Financial Assistance grants, while playing a vital role in supporting immunization "infrastructure," have been too unstable and unpredictable to elicit the strategic planning, programming, and own-source spending that would be optimal for state and local programs. The predominant immunization function of state and local public health agencies is becoming assurance of age-appropriate immunization throughout the lifespan. To be successful in this emerging role, the health agencies must be supported with appropriate staffing, interagency collaboration, and clearly articulated authority. PMID- 11024332 TI - Measuring immunization coverage. AB - OBJECTIVE: Information about immunization coverage comes from five major sources: the National Immunization Survey, the National Health Interview Survey, retrospective school-entry surveys, the Health Plan Employer Data and Information Set (HEDIS) measures reported by managed care plans, and assessments performed on clinics and private practices. In this article, we describe the methodology of the major surveys, discuss technical and policy issues in measuring immunization coverage, and identify issues that must be addressed to harmonize immunization rates calculated from different sources. METHODS AND TOPICS: We describe the (1) design and methodology of the five major sources of immunization coverage assessments, (2) issues and controversies in measuring immunization coverage, and (3) preliminary efforts to harmonize calculation of immunization coverage. Technical and policy issues involve dose and interval requirements, which vaccines are included in the series-completion calculations, and who is excluded from each method of calculation. CONCLUSIONS: The purpose of measuring up-to-date immunization coverage determines the way that it is measured. The tension between measuring immunization coverage to monitor population protection against disease and measuring immunization coverage to determine how well the health care delivery system is working leads to different ways of selecting a sample and reporting coverage. These differences create confusion for the public policymakers who try to identify problems and to set priorities for immunization efforts. Although some unavoidable differences may occur because of differences in purpose of the measurement, greater harmonization is possible. PMID- 11024333 TI - Immunization pockets of need: science and practice. AB - Despite high overall immunization coverage levels among U.S. preschool children, areas of underimmunization, called pockets of need, remain. These areas, which pose both a personal health and a public health risk, are typically poor, crowded, urban areas in which barriers to immunization are difficult to overcome and health care resources are limited. The purpose of this report is to review barriers to immunization of preschool children living in pockets of need and to discuss current issues in the identification of and implementation of interventions within these areas. The Centers for Disease Control and Prevention administers a federal grants program that funds state and metropolitan immunization programs. This program promotes a three-pronged approach for addressing pockets of need: (1) identification of target areas, (2) selection and implementation of programmatic strategies to improve immunization coverage, and (3) evaluation of progress or impact. At each step, scientific evidence can guide programmatic efforts. While there is evidence that state and metropolitan immunization programs are currently making efforts to address pockets of need, much work remains to be done to improve immunization coverage levels in pockets of need. Public health agencies must take on a broadened role of accountability, new partnerships must be forged, and it may be necessary to strengthen the oversight authority of public health. These tasks will require a concentration and redirection of resources to support the development of an immunization delivery infrastructure capable of ensuring the timely delivery of immunizations to the most vulnerable of America's children. PMID- 11024334 TI - Federal immunization policy and funding: a history of responding to crises. AB - BACKGROUND: This article outlines the history of federal immunization policy and funding, with a focus on discretionary federal funding under Section 317 of the Public Health Service Act, paying particular attention to the role of Congress in shaping the program in the past 2 decades. This review of funding trends and initiatives indicates that when both a presidential administration and key congressional actors viewed immunization as a priority and made sufficient funds available to support the public health delivery system and its infrastructure, coverage levels would continue to rise and disease levels continue to decline. From the beginning, immunization financing was explicitly structured as a federal state-private-sector partnership. Section 317 program's statute has not changed much in 35 years, despite significant changes to the health care delivery system, other federal immunization activities, and rates of immunization coverage. Although the creation and implementation of the Vaccines for Children (VFC) program in the mid-1990s resulted in some congressional deliberations over immunization policies, no explicit restructuring of the 317 program occurred as a result. The Section 317 program retains its traditional authority and mission to address urgent needs, sustain public delivery systems, and provide funds for purchase of vaccines. The question remains whether the resources to sustain progress in immunization can be secured during times with no crisis, to ensure constant "readiness" in immunization (as in defense), or whether another epidemic must occur before the federal government is willing to commit optimal resources. PMID- 11024335 TI - Breaking the "immunization cycle". Achieving and sustaining high levels of vaccine coverage will require eternal vigilance. PMID- 11024336 TI - Litter size, ovulation rate and prenatal survival in relation to ewe body weight: genetics review. AB - Lamb's birth weight and litter weight were found to be related to ewe weight. Deviations from generally applicable formulae indicated an interaction between the genotype of the lamb and the maternal environment, as suggested in egg transfer experiments. Land's (1977) hypothesis on the spare conceptus capacity, when comparing large and small breeds within a species, with respect to prolificacy, was more applicable to genetically homogeneous groups within a breed than between breeds and lines within a breed. A hypothesis suggests that variation in the within breed genetic correlation between ewe weight and prolificacy may largely depend on the relative uniformity of selection criteria applied. It can further be argued whether selection based on body weight or growth rate, compared to selection on reproductive performance, could be important sources of within breed as well as among breed variations in the genetic correlation estimates between ewe weight and prolificacy. The dynamics of the genetic correlation between ewe weight and prolificacy has an effect of long term selection for body weight, growth rate and reproduction traits. This leads to the problem of the relative incompatibility of selection of an association between body proportions with reproductive traits, especially in meat type breeds. There is a parallellism or antagonism in the genetic correlations between ewe weight and prolificacy as compared to those between ewe weight and other reproduction traits. The within breed relationship between ewe weight and ovulation rate was found to be positive in some breeds. The effects of body condition at mating were contradictory. Genotypexenvironment interactions were observed in certain cases. Among most breeds, differences in ewe weight were unrelated to differences in ovulation rate. There were no relationships between ewe weight and prenatal survival within breeds or in lines within a breed. However, there exists a differential relationship between ovulation rates and prenatal survival rates in relation to ewe weight, depending on the selection criteria applied. A clearcut relationship between litter weight components and ewe weight cannot be generalized but may vary among differentially selected breeds and lines within them. This is a final, unifying and unresolved question. PMID- 11024337 TI - Evaluation of Black Bengal goats and their cross with the Jamunapari breed for carcass characteristics. AB - Carcass quality of JamunaparixBlack Bengal (JBB), Black Bengal selected for growth (SBB) and random-bred (RBB) wethers of 1 year of age was studied. There was no significant difference between JBB and SBB in pre-slaughter traits and carcass characteristics, except height at wither and dressing percentage which were significantly higher in JBB than in SBB, and muscle fiber diameter which was significantly smaller in JBB than in SBB. Performance of RBB was significantly lower than JBB and SBB. When hot carcass, non-carcass, variety meat, prime cuts, fat deposition and total saleable portion were expressed as % of empty live weight and compared among genetic groups, JBB ranked first. Selection for rapid growth rate within Black Bengal goat increased its live weight and improved meat production. Increased live weight showed a positive relationship with increased fat deposition. Relationships between 'eye muscle' area and hot carcass yield, empty live weight and gut fat and perinephric fat and gut fat were positive and significant. PMID- 11024338 TI - Physiologic and biochemical effects of subarachnoidally administered xylazine and medetomidine in goats. AB - Clinicophysiological, haematological and biochemical effects of xylazine (0.05mgkg(-1)) and medetomidine (0.01mgkg(-1)) were studied in nine adult goats after lumbosacral subarachnoid administration. The onset of analgesia by xylazine and medetomidine was observed in 9.11+/-1.07 and 8.66+/-2.37min (mean+/-S.E.), respectively. Both alpha(2)-agonists produced moderate analgesia of hind quarter, perineum and flank, mild ataxia and sedation. The duration of analgesia after xylazine administration was 134.44+/-8.87min and that after medetomidine was 158.33+/-9.96min (mean+/-S.E.). Xylazine and medetomidine induced significant (p<0.05) decrease in heart rate, respiratory rate and hypothermia. Haemoglobin (Hb), packed cell volume (PCV) and total leukocyte count (TLC) decreased significantly. Changes in the physiological and haematological parameters were transient in nature. Xylazine and medetomidine produced a significant (p<0.05) increase in creatinine and glucose levels. However, these parameters fluctuated within normal range and started to recover within 120min. However, serum urea nitrogen (SUN), serum chloride, sodium and potassium did not show any significant change. The effects produced by xylazine and medetomidine were however, comparable at these dose levels. The study indicates that xylazine at 0.05mgkg( 1) and medetomidine at 0.01mgkg(-1) did not induce any serious alteration in the physiological, haematological and biochemical parameters and can be safely used in inducing hind quarter, flank and perineal analgesia in goats. PMID- 11024339 TI - Chronic toxicity of arsenic in goats: clinicobiochemical changes, pathomorphology and tissue residues. AB - Chronic inorganic arsenic toxicity was induced in goats by oral administration of one-fifth of the acute lethal dose 50 (ALD(50)) of sodium arsenite (25mgkg(-1) body weight) packed in gelatin capsules and given daily for 12 weeks. Clinical signs of toxicity developed from 3 week post-exposure, consisting of gastrointestinal disturbances and renal insufficiency with 100% mortality in all animals. There were significant (p<0.01) decreases in total serum protein and the albumin: globulin ratio, and increases in blood glucose and various enzymatic activities of treated animals. Toxicity also induced severe pathomorphological changes, indicative of haemorrhagic and degenerative and/or necrotic lesions in most organs. In addition, proliferative pneumonia in lungs, hyperplastic goitre in thyroid and chronic proliferative lesions in skin were observed. Liver contained the largest residues of arsenic, followed by intestine, kidneys, thyroid, abomasum, spleen, skin, lungs and lowest in brain. The intensity of pathomorphological changes was proportional to the accumulated amount of arsenic in tissues/organs. PMID- 11024340 TI - Effects of shearing and supplemental level on intake of dry ewes grazing on barley stubble. AB - Twenty-eight Comisana dry ewes were utilised to evaluate the effect of shearing (14 shorn and 14 unshorn ewes) and supplemental level (100 and 300g of concentrate) on intake of barley stubble. Shorn ewes showed a small increase in dry matter intake (at the end of August: 1.32 vs. 1.10kg/day; p<0.01). Intake of barley stubble decreased as supplement level increased (mean: 1.18 vs. 1.29; p<0.01). On average, substitution effect was 0.63. Total lamb weight per ewe was greater in the group receiving 300g of concentrate (7.38 vs. 5.85; p<0.05). Shearing induced a small, but not significant decrease in total lamb weight per ewe (6.18 vs. 7.05kg/ewe). Results seem to indicate that unshorn ewes react better to environmental stress and that 300g improve reproductive performance. PMID- 11024341 TI - Estrus synchronization in ewes treated with sponges impregnated with different doses of medroxyprogesterone acetate. AB - The objectives of this study were to determine residual and absorbed levels of medroxyprogesterone acetate (MAP) after treatment with intravaginal sponges impregnated with different doses of MAP in an ewe herd and to determine the effects of such treatments upon estrus incidence, interval to estrus onset and pregnancy rate. Polyurethane sponges impregnated with 40, 50 and 60mg of MAP were prepared. Real amounts of progestagen on sponges were checked prior to sponge treatment insertion. During autumn 1999, 608 cyclic Merino ewes were treated with intravaginal sponges impregnated with different doses of MAP (groups I, 40; II, 50 and III, 60mg). After 14 days, sponges were removed and residual levels of MAP (RMAP) on removed sponges were measured by spectrophotometry. Real amounts for unused sponges were 39.60+/-1.10, 44.10+/-1.06 and 59.10+/-1.26mg for the intended doses of 40, 50 and 60mg MAP respectively. RMAPs were different (p<0.05) among groups (I: 17.98+/-1.97mg; II: 24.32+/-2.03mg; III: 34.25+/-3.23mg). Absorbed levels of MAP were not different among groups (I: 21.62+/-1.97mg; II: 19.78+/-2.03mg; III: 24.85+/-3.23mg). Artificial insemination was performed in 16 ewes from Group I, 17 ewes from Group II and 11 ewes from Group III, 12h after estrus onset. There were no differences among groups, neither for estrus incidence (I: 79.27%; II: 77.42%; III: 80.87%) nor interval to estrus onset (I: 55.94+/-1.87h; II: 56.74+/-1.13h; III: 57.70+/-1.02h). There were also no differences among groups for pregnancy rate (I: 43.75%; II: 52.94%; III: 45.45%). It was concluded that under similar conditions, a dose as low as 40mg MAP could be effectively used for estrus synchronization in cyclic Merino ewes. PMID- 11024342 TI - Macromineral profile in the plasma of Nubian goats as affected by the physiological state. AB - A total of 65 Nubian dairy goats were selected to represent a combination of age and physiological status. Changes in some plasma concentrations underlying different physiological states were monitored. There were young and adult animals, pregnant animals, all with more than 50 days after kidding, divided into subgroups according to gestation, first, second or third pregnancy. Lactating animals up to 50 days after kidding were divided into subgroups according to parity (first, second or third) and stage of lactation (early or mid). The last category consisted of animals grouped according to yield (high or low). Data analyses have shown that calcium (Ca) levels were lowest (P<0.05) during first early lactation, while those of phosphorous (P) were lowest (P<0.05) in second pregnancy and also in early lactations compared with mid-lactations (P<0.05). Magnesium (Mg) levels increased (P<0.05) with age and number of pregnancies. The order was second, third and first. Irrespective of parity, mid-lactations showed higher (P<0.05) Mg levels than early ones. Sodium (Na) plasma levels decreased (P<0.05) progressively with age and from first to third pregnancy; and decreased (P<0.05) during first and third mid-lactations compared with first or second early lactation, and as well as with second mid-lactation. Potassium (K) plasma levels were lower (P<0.05) in early than mid-lactations. Except for Mg, the type of milk yield had no significant effect on minerals under study. Minerals in the diet of animals kept under different management practices must be regarded among other essential nutrients. Physiological status might modify its requirement to these elements. PMID- 11024343 TI - Adrenocorticotrophic hormone and dexamethasone failed to affect milk yield in dairy goats: comparative aspects. AB - The ability of adrenocorticotrophic hormone (ACTH; single i.v. injection of 2.5IU/kg BW) and dexamethasone (single i.m. injection of 36mg/kg BW) to affect milk production was studied in mid-lactating Israeli Saanen goats. None of these treatments produced changes in milk yield and composition of the goats. The effects of ACTH on blood cortisol levels, and the effects of ACTH and dexamethasone on blood plasma concentrations of glucose, however, were consistent with previous reports in goats and cows. These responses suggest that ACTH and dexamethasone treatments produced their expected glucocorticoid effects. It is suggested that obstructing the axis: stress-ACTH-glucocorticoid-down regulation of milk yield, which was demonstrated in dairy cows, reflects the adaptation of goats to harsh conditions, and the selection pressure to produce milk under conditions which are considered stressful for other ruminants. PMID- 11024344 TI - Erratum to "Pharmacokinetics and penetration into tissue fluid of ceftizoxime in normal and hyperthemic sheep". PMID- 11024345 TI - Salmonella enteritidis temperature-sensitive mutants protect mice against challenge with virulent Salmonella strains of different serotypes. AB - The protection conferred by temperature-sensitive mutants of Salmonella enteritidis against different wild-type Salmonella serotypes was investigated. Oral immunization with the single temperature-sensitive mutant E/1/3 or with a temperature-sensitive thymine-requiring double mutant (E/1/3T) conferred: (i) significant protection against the homologous wild-type Salmonella strains; (ii) significant cross-protection toward high challenge doses of S. typhimurium. Significant antibody levels against homologous lipopolysaccharide and against homologous and heterologous protein antigens were detected in sera from immunized mice. Moreover, a wide range of protein antigens from different Salmonella O serotypes were recognized by sera from immunized animals. Besides, primed lymphocytes from E/1/3 immunized mice recognized Salmonella antigens from different serotypes. Taken together, these results indicate that temperature sensitive mutants of S. enteritidis are good candidates for the construction of live vaccines against Salmonella. PMID- 11024346 TI - The study of cell-mediated immune response in recurrent vulvovaginal candidiasis. AB - The aim of this work was to examine in vitro the ability of cells from patients with recurrent vulvovaginal candidiasis (RVVC) to cell-mediated immune response. Peripheral blood mononuclear cells (PBMC) and whole blood cells (WBC) of 37 RVVC patients in acute infection and 14 in remission were examined for the ability to proliferation and cytokines production (IFN, TNF, IL-6). As a control, a group of 25 healthy women were examined. The cells were stimulated with Candida antigen (HKCA), LPS and PHA. To indicate the level of cytokines, the following cell-lines were used: A549 for IFN, WEHI 164 for TNF and 7TD1 for IL-6. The proliferation/death of cells was determined by colorimetric test using MTT. Distinct suppression of cell-mediated immune response (CMI) was shown in all patients comparing to the control. Greatest suppression was found in the acute phase of the disease. The ability of cells to proliferate and produce IFN increases only in remission. The data seem to suggest that in this phase of disease, the ability of cell-mediated immune response is restored. It was also indicated that IFN may take part in protection against Candida infection. PMID- 11024347 TI - Detection of microsporidia in surface water: a one-year follow-up study. AB - In order to estimate the rate and seasonal variation of Enterocytozoon bieneusi contamination of surface water, sequential samples of water from the River Seine in France were collected during a 1-year period. Each sample (300-600 l) was submitted to sequential filtrations, and the filters were then examined for microsporidia using light microscopy and nested polymerase chain reaction (PCR) for E. bieneusi. Amplified products were hybridized with a E. bieneusi-specific probe. Twenty-five samples of water were analyzed during 1 year. Microscopic examination of stained filters proved unreliable for the identification of spores. Using nested PCR, 16 of 25 specimens were positive (64%). Unexpectedly, E. bieneusi was identified in only one sample by specific hybridization underlining the lack of specificity of ours primers. Nevertheless, using DNA sequence analysis, unknown microsporidia species were identified in eight cases, which had highest scores of homology with Vittaforma corneae or Pleistophora sp. This study shows a low rate of water contamination by E. bieneusi suggesting that the risk of waterborne transmission to humans is limited. PMID- 11024348 TI - Effect of Alloiococcus otitidis and three pathogens of otitis media in production of interleukin-12 by human monocyte cell line. AB - Alloiococcus otitidis is detected in middle ear effusion of otitis media with effusion (OME). Only a limited number of studies are available concerning the immunological profile of A. otitidis. We have studied the ability of A. otitidis and three other representative pathogens of otitis media to stimulate the production of interleukin-12 (IL-12) from a monocytic cell line THP-1. Viable A. otitidis induced the production of IL-12 in THP-1 cells but IL-12 production was reduced if glutaraldehyde-fixed bacteria were used as stimulants. When viable bacteria were physically separated from THP-1 cells during the stimulation period, remarkable reductions of IL-12 secretion were shown after challenge with gram-positive bacteria A. otitidis and S. pneumoniae. When stimulated with soluble extracts of A. otitidis, THP-1 secreted IL-12 in a dose-dependent manner. The subfraction with a molecular mass over 100 kDa showed a strong ability to induce IL-12 production. Our results show that A. otitidis has immunostimulatory capacity with regard to IL-12 production. We also show that soluble antigen(s) of A. otitidis can modulate the immune response in OME. PMID- 11024349 TI - Specific detection of Plesiomonas shigelloides isolated from aquatic environments, animals and human diarrhoeal cases by PCR based on 23S rRNA gene. AB - Twenty-five strains of Plesiomonas shigelloides isolated from aquatic environment, 10 strains from human cases of diarrhoea and five strains from animals were identified by the polymerase chain reaction technique based on 23S rRNA gene. For this purpose, two primers targeted against part of the 5' half of the 23S rRNA gene of P. shigelloides (Escherichia coli number C-912, G-1195; Plesiomonas number C-906, G-1189) were designed. Results from our study indicated that this method might serve as a tool for a rapid and sensitive identification of P. shigelloides from different environmental and clinical sources. PMID- 11024350 TI - Identification of genes associated with copper tolerance in an adhesion-defective mutant of Aeromonas veronii biovar sobria. AB - TnphoA mutagenesis was used to identify adhesins of Aeromonas veronii biovar sobria 3767, a strain isolated from a diarrhoeal stool specimen. Six mutants, from a library of 154, exhibited significantly reduced levels of adhesion to HEp 2 cells. Primers to the terminal regions of TnphoA were used for inverse PCR and the product from one mutant was cloned into pBluescript and partial sequence data obtained. Scanning GenBank and EMBL data bases revealed DNA sequence similarity to the copA gene of Pseudomonas syringae pv. tomato which confers resistance to copper and other heavy metals. The transposon was located within the copA gene and the mutant exhibited a reduced tolerance to copper. Primer walking, using the inverse PCR product as a template, revealed three open reading frames (ORFs) copA, B and C in A. veronii biovar sobria 3767. The predicted amino acid sequences of ORFs A and B had significant homology (55 and 34% respectively) to the copA and B proteins of P. syringae. No amino acid or DNA sequence homology existed between ORF C of strain 3767 and any other gene in the data bases scanned. Further analysis of the nucleotide sequence failed to reveal the presence of typical copper regulatory genes within the vicinity of the Aeromonas sequence. The association between copper tolerance and adhesion in A. veronii biovar sobria requires further study. PMID- 11024351 TI - Candidate multi-epitope vaccines in aluminium adjuvant induce high levels of antibodies with predefined multi-epitope specificity against HIV-1. AB - Some neutralizing epitopes on HIV-1 envelope proteins were identified to induce antibodies which could effectively inhibit the infection of different strains in vitro. But only very low levels of these antibodies were determined in the HIV-1 infected individuals. To increase the levels of protective antibodies in vivo, we suggested multi-epitope vaccine as a new strategy to induce high level of neutralization antibodies with predefined multi-epitope specificity. A synthesized epitope peptide MP (CG-GPGRAFY-G-ELDKWA-G-RILAVERYLKD) containing three neutralizing epitopes (GPGRAFY, ELDKWA, RILAVERYLKD) was conjugated to carrier protein KLH, and then used for immunization in mouse together with aluminium adjuvant or Freund's adjuvant (FA). The candidate MP-KLH multi-epitope vaccine in aluminium adjuvant could induce antibody response very strongly to the epitope peptide C-(RILAVERYLKD-G)2 and the immunosuppressive peptide (P1) (LQARILAVERYLKDQQL) (antibody titer: 1:51200), strongly to the epitope peptide C (ELDKWA-G)4 and the C-domain peptide (P2) (1:12800), and moderately to the epitope peptide C-(GPGRAFY)4 and the V3 loop peptide (1:1600). The immunoblotting analysis demonstrated that the antibodies in sera could recognize P1, P2, V3 loop peptides and rsgp41 (aa 539-684). These results are similar with that in the case of PI-BSA in FA, and suggest that the multi-epitope vaccine in aluminium could induce high levels of antibodies of predefined multi-epitope specificity, which provides experimental evidence for the new strategy to develop an effective neutralizing antibody-based multi-epitope vaccine against HIV-1. PMID- 11024352 TI - Role of porin of Shigella dysenteriae type 1 in modulation of lipopolysaccharide mediated nitric oxide and interleukin-1 release by murine peritoneal macrophages. AB - The ability of Shigella dysenteriae type 1 porin to induce the release of nitric oxide (NO) and interleukin-1 (IL-1) from peritoneal macrophages of mouse and to regulate lipopolysaccharide (LPS) and gamma interferon (IFN-gamma) mediated release of the two proinflammatory mediators was investigated. Porin released nitrite when added to macrophage cultures. A maximum of 3.2-fold nitrite release by macrophages was observed with 100 ng ml(-1) of porin. The nitrite release of LPS was enhanced significantly by lower concentrations of porin, whereas the effect of IFN-gamma was enhanced by porin at higher concentrations. Polysaccharide (PS) moiety of LPS stimulated the nitrite release of elicited macrophages by 1.6-fold compared to untreated control. It also enhanced the stimulatory effect of 1 and 10 ng ml(-1) of porin by 1.3-fold. Lipid A (LPA) moiety of LPS did not release nitrite, nor did it increase the porin mediated nitrite production. Porin treated 24 h old macrophage culture supernatants were applied for ConA activated thymocyte proliferation as a measure for determination of IL-1 release. Sixty percent depletion of thymocyte proliferation was observed when the porin treated macrophage supernatants were absorbed with anti-IL-1 antibody. A maximum of 5.5-fold increase of thymocyte proliferation over control was found with 1 and 10 ng ml(-1) of porin. One or 10 ng ml(-1) of porin and LPS augmented the thymocyte growth, 1.5-fold beyond that obtained by porin and 1.8 /1. 7-fold more than that obtained by LPS, alone. Similarly, porin and IFN-gamma co-stimulated the cell growth also. PS enhanced the thymocyte proliferation by 5 fold. It also enhanced the thymocyte growth by co-stimulating 1.4-fold the effect observed by 1 or 10 ng ml(-1) of porin alone. LPA could not participate in the cell proliferating activity nor did it enhance the stimulatory effect of porin. Therefore, both nitrite release and thymocyte proliferation by LPS could be substituted by PS only. The tight association of the two bacterial outer membrane components, porin and LPS, could be a necessary co-signal for boosting the release of the two proinflammatory mediators, namely NO and IL-1, which may be associated with the inflammatory response of the colon during Shigella invasion. PMID- 11024353 TI - Virulence of a Proteus mirabilis ATF isogenic mutant is not impaired in a mouse model of ascending urinary tract infection. AB - Proteus mirabilis, a common cause of urinary tract infection, produces a number of different fimbriae, including ambient temperature fimbriae (ATF). These fimbriae are optimally expressed at 23 degrees C and their contribution to urinary tract infection has so far remained unknown. In the present study, a clinical isolate of P. mirabilis and an isogenic allelic replacement mutant unable to express ATF were tested for their ability to cause infection in the ascending urinary tract infection model in mice. The atf mutant colonised the urinary tract as well as the wild-type strain and was also able to outcompete the wild-type strain in a co-challenge experiment. Different non-clinical P. mirabilis isolates showed a reactive AtfA band after Western blot analysis using a polyclonal rabbit AtfA antiserum. These data together suggest that ATF does not play a role in P. mirabilis urinary tract infection. PMID- 11024354 TI - Human antibody response during sepsis against targets expressed by methicillin resistant Staphylococcus aureus. AB - The identification of target structures is a prerequisite for the development of new treatment options, like antibody based therapy, against methicillin resistant Staphylococcus aureus (MRSA). In this study we identified immunodominant structures which were expressed in vivo during sepsis caused by MRSA. Using human sera we compared the immune response of humans with MRSA sepsis with the immune response of normal individuals and asymptomatically colonized individuals. We identified and characterized four staphylococcal specific antigenic structures. One target is a staphylococcal protein of 29 kDa that exhibited 29% identity to secreted protein SceA precursor of Staphylococcus carnosus. The putative function of this protein, which was designated IsaA (immunodominant staphylococcal antigen), is unknown. The second target is an immunodominant protein of 17 kDa that showed no homology to any currently known protein. This immunodominant protein was designated IsaB. The third and fourth antigens are both immunodominant proteins of 10 kDa. One of these proteins showed 100% identity to major cold shock protein CspA of S. aureus and the other protein was identified as the phosphocarrier protein Hpr of S. aureus. The identified immunodominant proteins may serve as potential targets for the development of antibody based therapy against MRSA. PMID- 11024355 TI - An epidemiological study on the occurrence of Staphylococcus aureus in superficial abscesses of patients presenting for surgery in a teaching hospital in Khartoum, Sudan. AB - A group of patients (n=86) suffering from superficial abscesses was recruited in the Khartoum Teaching Hospital, Sudan. Detailed clinical and socio-economic data were collected. It appeared that 83% of all patients were younger than 40. Labourers were most prevalent (28%), followed by students (23%) and housewives (16%). The head and neck were most often affected (22%), with hands being second (19%). In 92% of all pus cultures a microbial agents was identified, the large majority being Staphylococcus aureus (69%). Among patients, 47% were nasal carriers of S. aureus, similar to the carriage rate measured among controls, suggesting that nasal carriage is no risk factor for abscess development. Multivariate logistic regression analysis revealed that a history of abscess, recent traditional medical treatment, poor hygiene and low socio-economic status were significantly and independently associated with the occurrence of superficial abscesses. PMID- 11024356 TI - Expression vectors and delivery systems. Food BiotechnologyWeb alert AB - A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Biotechnology. PMID- 11024357 TI - Expression vectors and delivery systems. Playing alien genes in remote theaters PMID- 11024358 TI - Surface-induced and biofilm-induced changes in gene expression. AB - A biofilm is a community of microorganisms attached to a surface. Based on studies of single-species communities, biofilm formation follows a progression from initial attachment to a mature form composed of pillar-like multicellular structures interspersed with fluid-filled channels. The developmental progression leading to a mature biofilm requires changes in gene expression. With recent technological advances for visualizing biofilm growth, gene expression can be directly monitored during biofilm development. Hence, analyses of surface-induced and biofilm-induced changes in gene expression have begun in earnest. Recent studies have identified regulatory pathways that are important for biofilm formation and have focused on genetic responses to environmental stimuli in mature biofilms. These findings are providing new insights into biofilm development and physiology. PMID- 11024359 TI - Detection of genes essential in specific niches by signature-tagged mutagenesis. AB - Variations of the signature-tagged mutagenesis (STM) technique are now possible and the method can be applied to most pathogens that have an STM-selectable phenotype in a host system. STM screening of 15,040 mutants from 11 bacterial species identified 323 in vivo attenuated mutants. As a genome-scanning tool, STM will yield information about genes with unknown functions as well as information crucial for understanding microbial pathogenesis. PMID- 11024360 TI - In vivo expression technology strategies: valuable tools for biotechnology. AB - Whole genome sequences have shown that bacteria possess a significant number of genes that have no known function. It is probable that many of these are required for survival in environments other than the agar plate. In vivo selection strategies provide a means of obtaining genes active in complex natural environments. Direct access to these genes is essential for understanding ecological performance and provides novel opportunities for biotechnology. PMID- 11024361 TI - The use of recombinant antibodies in proteomics. AB - Recombinant antibodies are becoming increasingly important in the field of proteomics. Recent advances include the development of large phage-antibody libraries that contain high-affinity binders to almost any target protein, and new methods for high-throughput selection of antibody-antigen interactions. Coupled with a range of new screening technologies that use high-density antibody arrays to identify differentially expressed proteins, these antibody libraries can be applied to whole proteome analysis. PMID- 11024362 TI - Expressing genes in different Escherichia coli compartments. AB - Production of heterologous proteins or parts thereof in different extra cytoplasmic compartments (in the periplasm, outer membrane or extracellularly) of Escherichia coli offers multiple applications, for example, in vaccine development, immobilised enzymes and bioremediation. Nowadays, not only surface display of short peptides, but also cell-surface anchoring or secretion of functional proteins is possible. Factors influencing folding, stability and export of extra-cytoplasmic proteins are also better understood. PMID- 11024363 TI - Site-specific gene targeting for gene expression in eukaryotes. AB - Major advances in the use of site-specific recombinases to facilitate sustained gene expression via chromosomal targeting have been made during the past year. New tools for genomic manipulations using this technology include the discovery of epitopes in recombinases that confer nuclear localization, crystal structures that show the precise topology of recombinase-DNA-substrate synaptic complexes, manipulations of the DNA recognition sequences that select for integration over excision of DNA, and manipulations that make changes in gene expression inducible by drug administration. In addition, endogenous eukaryotic and mammalian DNA sequences have been discovered that can support site-specific recombinase mediated manipulations. PMID- 11024364 TI - Translocating peptides and proteins and their use for gene delivery. AB - A dramatic surge in the development of peptides for gene delivery in vitro and in vivo has been witnessed in the past decade. A better understanding of the structural and mechanistic properties of peptides has been an important step for the rational design of optimal peptide-based gene delivery systems. Research has focused on the design of short synthetic peptides that overcome both extracellular and intracellular limitations of other gene delivery systems by binding reversibly and condensing DNA, specifically targeting cells and/or tissues, rapidly releasing plasmids into the cytoplasm and mediating efficient nuclear translocation. PMID- 11024366 TI - Food biotechnology. Editorial overview PMID- 11024365 TI - Bacterial promoters triggering biodegradation of aromatic pollutants. AB - Unraveling the complex transcriptional regulation of bacterial catabolism of aromatic pollutants is a prerequisite for engineering efficient biological systems for many biotechnological applications. A first level of regulation relies on specific regulator-promoter pairs. There have been new insights into the molecular mechanisms that regulatory proteins use to sense a given signal and to activate transcription initiation from the cognate promoters. A second level of regulation allows adjustment of the expression of the particular catabolic operons in response to the global environmental conditions of the cells, and recent findings provide some clues about the mechanisms underlying such complex regulatory checkpoints. PMID- 11024367 TI - Filamentous fungi as microbial cell factories for food use. AB - The knowledge base that will underpin the more efficient use of filamentous fungi as cell factories in food has increased during the past year in the areas of gene regulation, protein secretion, safety and synthesis of ingredients such as long chain polyunsaturated fatty acids. PMID- 11024368 TI - Biotechnological approaches to the understanding and improvement of mature cheese flavour. AB - There have been important milestones in biotechnological practice that have led to the determination and production of superior cheese flavours. Within the past year, the use of gas chromatographic techniques and sensory methodologies has been optimised by several groups in efforts to evaluate the organoleptic properties of a number of mature cheeses. The hydrolysis of milk caseins, small peptides, free amino acids and fatty acids, and the generation of sulfur containing compounds are uniformly assumed to result in the formation of specific cheese aromas. Giant strides have been taken in molecular technology to aid the dissection and exploitation of the metabolic pathways that lead to the formation of these flavour constituents. Specific advances in molecular technology have included metabolic engineering of lactic acid bacteria for enhanced flavour development. PMID- 11024369 TI - Novel approaches to the biosynthesis of vanillin. AB - Microorganisms able to produce vanillin in excess of 6g/l from ferulic acid have now been isolated. In Pseudomonas strains, the metabolic pathway from eugenol via ferulic acid to vanillin has been characterised at the enzymic and molecular genetic levels. Attempts to introduce vanillin production into other organisms by genetic engineering have begun. PMID- 11024370 TI - Advances in the genetics of thermophilic lactic acid bacteria. AB - Molecular genetics of thermophilic lactic acid bacteria has advanced in several directions: exploitation of the milk proteins and sugars; primary and secondary metabolism; stress response; and molecular ecology of bacteria and their phages. These have singularly contributed to open new avenues of scientific interest in the field: comparative phage genomics; horizontal gene transfer events in bacterial or phage populations; and genetics of external polysaccharide production. PMID- 11024371 TI - Gene transfer from genetically modified food. AB - The current debate about the safety of genetically modified food includes some important scientific issues where more scientific data would aid the robustness of safety evaluation. One example is the possibility of gene transfer, especially from genetically modified plant material. PMID- 11024372 TI - European criteria for the appropriateness and necessity of coronary revascularization procedures. AB - OBJECTIVES: Large variations in the use of coronary revascularization procedures have led many countries to apply the RAND appropriateness method to develop specific criteria describing patients who should be offered these procedures. The method is based on the work of a multidisciplinary expert panel that reviews a synthesis of the scientific evidence and rates the appropriateness of a comprehensive list of indications for the procedure being studied. Previous studies, however, have all involved single-country panels. We tested the feasibility of carrying out a multinational panel to rate the appropriateness and necessity of coronary revascularization, thereby producing recommendations for common European criteria. METHODS: Using the RAND methodology, a multispecialty (interventional cardiologists, non-interventional cardiologists and cardiovascular surgeons), multinational (The Netherlands, Spain, Sweden, Switzerland and the United Kingdom) panel rated the appropriateness and necessity of indications for percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass graft surgery (CABG). A synthesis of the evidence and list of indications for PTCA and CABG were sent to 15 panelists, three from each country, who performed their ratings in three rounds. RESULTS: For PTCA, 24% of the indications were appropriate and necessary, 16% were appropriate, 43% were uncertain and 17% were inappropriate. The corresponding values for CABG were 33% appropriate and necessary, 7% appropriate, 40% uncertain and 20% inappropriate. The proportion of indications rated with disagreement was 4% for PTCA and 7% for CABG. CONCLUSION: Multinational panels appear to be a feasible method of addressing issues concerning the appropriateness and necessity of medical procedures in western European countries. The criteria produced provide a common tool that can be used to measure the overuse and underuse of medical procedures and to guide decision-making. PMID- 11024373 TI - Reduced risk of coronary artery bypass surgery for unstable angina during a 6 year period. AB - OBJECTIVE: To assess the risk of early death or acute myocardial infarction in patients undergoing isolated coronary artery bypass surgery for unstable coronary artery disease. METHODS: Retrospective observational study of 853 patients operated on because of unstable coronary artery disease during 1990-1995. RESULTS: There were 5.9% deaths and 13.0% nonfatal infarctions < or =30 days. These figures declined during the observation period and were 2.6% and 6.2%, respectively, in 1995. The relative risk of early death or myocardial infarction was 50% less during 1994-1995 than during 1990-1991, after multivariate adjustment for several patient risk factors. The risk of death or infarction was almost twice as high in patients > or =50 years than in those < 50 years of age. Multivariate analysis showed that aortic-cross-clamp time > or =60 min, previous bypass surgery, pre-operative heart failure, emergency surgery, worse Braunwald class and non-use of an internal mammary artery graft were associated with an increased risk of death or infarction. Early mortality was 3.4% (24/702) in unstable patients without symptoms of congestive heart failure, who were not operated on emergently after failed percutaneous coronary intervention and had not had previous cardiac surgery. CONCLUSIONS: We observed a marked reduction of the risk of early death or myocardial infarction after surgery for unstable angina during the 6-year period 1990-1995. The risk reduction was not explained from operations performed on patients with less risk and indicates improved peri operative patient management. PMID- 11024374 TI - Aortocoronary bypass grafting: a comparison of HTK cardioplegia vs. intermittent aortic cross-clamping. AB - OBJECTIVE: Intermittend, hypothermic aortic cross-clamping (IAC) with myocardial fibrillation and cardioplegic arrest (CA) have been established both as effective methods for coronary artery bypass surgery (CABG). Nevertheless, there exists controversy about the more beneficial cardioprotective effect of one of these procedures in CABG-patients. METHODS: In this prospective study we compared the clinical outcome, ischemic serum-markers (CK, CK-MB, Troponin I), electrocardiogram (ECG)-changes, and hemodynamic data of 103 patients. Randomization in group I (IAC; n=52) or group II (CA; n=51) was done consecutively, all data were compared by Student's t-test or chi(2)-test and P<0.05 was regarded as significant. The Bretschneider-HTK solution was used for cardioplegic arrest. Data were collected before operation, before ischemic arrest, after 5 and 60 min of reperfusion, 1 and 6 h after operation, 1, 2 and 10 days postoperatively. RESULTS: There were no significant differences between both groups regarding general patient data: age (IAC: 64. 8+/-9.2 vs. CA: 63.8+/-9.0 years), left ventricular function (ejection fraction: IAC: 62+/-14 vs. CA: 64+/ 13%), the amount of bypassed vessels (IAC: 3.4+/-0.5 vs. CA: 3.6+/-0.5), total bypass time (IAC: 113+/-31 vs. CA 108+/-20 min). The total time of ischemia was significantly less in the IAC group with 37+/-10 vs. 48+/-10 min in the CA group. In the IAC-group, a higher mortality was noticed (7. 7 vs. 3.9%; N.S.). This was combined with a significantly higher amount of patients with peak serum-values of CK-MB (>40 U/l) and troponin I (>50 ng/ml), 17 in the IAC-group (33%) vs. eight in CA-group (16%). Cerebral strokes were seen in two IAC-patients and none in CA patients (NS). ECG-changes occurred in 22 IAC patients (42%) vs. 16 CA patients (31%); persistent ischemia related ECG-changes in six IAC (11.5%) vs. five CA patients (9.8%). CONCLUSIONS: Both cardioprotective methods, IAC and HTK cardioplegia, seem to offer sufficient myocardial protection in normal CABG procedures. Although neurologic disorders and mortality rates were higher in patients with intermittent aortic cross-clamping, the differences to the cardioplegia group were not significant. According to the analysis of increased ECG-changes, higher CK-MB and troponin I values, which occurred especially in patients with myocardial ischemia time longer than 40 min, we conclude that cardioplegic arrest with HTK seems to offer more beneficial effects in procedures with prolonged ischemia. PMID- 11024375 TI - Endothelial function of human vena saphena magna prepared with different minimally invasive harvesting techniques. AB - OBJECTIVES: Minimally invasive saphenous harvesting techniques have been shown to reduce post-operative morbidity. However, when applying new techniques of vein harvesting, endothelial integrity should be preserved in order to guarantee graft quality. We investigated the impact of two different minimally invasive saphenous vein harvesting techniques on endothelial function compared with the traditional 'open' technique by inducing endothelium-dependent in vitro relaxation. METHODS: Two different minimally invasive techniques for harvesting the greater saphenous vein were used in 66 patients, either using a video-assisted dissector (ETHI, n=33) or a light coupled retractor (AUTS, n=33); other patients were treated conventionally (CONV, n=30). The physiological response was tested in vitro using an organ chamber on vein segments exposed to acetylcholine after precontraction with potassium. RESULTS: In both minimally invasive groups, two patients each had to be converted to the traditional open technique and dropped out of the study. The endothelial function of the other veins harvested by either of the minimally invasive techniques showed no significant difference compared with veins harvested conventionally. Reactivity was measured as the percentage relaxation of pharmacological contraction and was significant in all groups (P<0.05); ETHI, 49.1+/-4.2%; AUTS, 48.8+/-5.1%; and CONV, 51.7+/-6. 0%. The responder/non responder ratio was similar in all groups: ETHI (two drop-outs), 28/3 (90.3%); AUTS (two drop-outs), 28/3 (90. 3%); and CONV, 27/3 (90.0%). CONCLUSION: Veins harvested by minimally invasive techniques have not shown an impaired reactivity of the endothelium compared with the conventional technique. PMID- 11024376 TI - Effect of transmyocardial laser revascularization on chronic ischemic hearts in sheep. AB - BACKGROUND: We investigated the effect of transmyocardial laser revascularization (TMR) on myocardial function and regional blood flow in an animal model of ischemic heart disease. METHODS: Chronic ischemia was induced in 11 sheep by the application of coronary stenosis on the left anterior descending (LAD) and circumflex coronary artery (LCX). Ten weeks later, in six of them, transmyocardial channels were created in the anterior free wall and in the posterior wall of the left ventricle. Five animals served as controls. The myocardial function was assessed by echocardiography taken at baseline and every 2 weeks after coronary stenosis and after TMR. Myocardial perfusion was measured by colored microspheres, injected at baseline, immediately after coronary stenosis, before and after TMR, and at 20 weeks after coronary stenosis. The hearts were retrieved at 20 weeks for light microscopic examination. RESULTS: The left ventricular end-diastolic and end-systolic cavity area was elevated 20 weeks after coronary stenosis in the control and TMR groups. There was no difference between groups (analysis of variance; ANOVA, non-significant). The wall thickening fraction (WTF) decreased progressively and significantly after coronary stenosis in both groups. The WTF was further acutely reduced by TMR, and recovered gradually to the pre-TMR level. No significant difference in WTF was observed between the TMR and control groups. The resting myocardial blood flow was significantly increased by TMR at 20 weeks (P=0.03). Light microscopic examination revealed channel patency in 49% of the laser scars at 10 weeks post TMR. A dense capillary network was observed at the edges of the surrounding scar. CONCLUSIONS: In an experimental model of ischemic heart disease, TMR developed angiogenesis in the lased channels, but, however, failed to improve myocardial function. PMID- 11024377 TI - The influence of age on gastro-oesophageal reflux: a re-appraisal of the DeMeester scoring system. AB - OBJECTIVE: The statistical evaluation of the influence of age on the pattern of gastro-oesophageal reflux (GOR) identified by prolonged pH monitoring in asymptomatic subjects. Re-appraisal of the DeMeester scoring system for GOR. METHOD: Prolonged pH monitoring was performed on 45 asymptomatic elderly adults with normal contrast oesophagogram, manometry and endoscopy. They included 36 males and nine females. The mean age was 66.6 years. The monitoring time ranged between 20 and 24 h, including one complete daily feeding cycle. GOR was defined as a reflux event with a pH of <4. The mean, standard deviation, kurtosis and skew were calculated for six parameters listed in the results. The mean values were compared with those of normal values determined by DeMeester et al. (in: Read NW, editor. Gastrointestinal motility: which test? 1989, pp. 43-52) from their study of 50 young healthy adults, and the t-test was applied to determine the statistical significance of differences. The 'null hypothesis' for each parameter was defined as 'the mean values of the elderly population are not statistically different from DeMeester's normal values'. RESULTS: The means (+/ SD) of six parameters studied in pH monitoring were as follows: supine reflux time as a percentage of total study time, 2.94+/-5.18%; upright reflux time as a percentage of total study time, 4.14+/-5.71%; total reflux time as a percentage of total study time, 3.5+/-4.38%; duration of longest reflux episode, 14.98+/ 24.92 min; number of reflux episodes lasting >5 min, 1.76+/-2.75; total number of reflux episodes during study, 13.49+/-11.31. These results were significantly different from the normal values reported by DeMeester. In addition, the data for each individual parameter was grossly skewed, as well as kurtotic, which implied that the data did not represent a normally distributed population. Moreover, we believe that the equation used for calculation of the DeMeester score, is inappropriate. CONCLUSIONS: The null hypothesis is rejected as the mean values of these parameters in our group are significantly higher than those used as normal. This implies that the normal values defined by DeMeester would over-diagnose gastro-oesophageal reflux disease (GORD). Moreover, we have found that the formula used to calculate the DeMeester's score is not according to the principle it is based on. DeMeester's system scoring is therefore inappropriate. PMID- 11024379 TI - [(18)F]Fluorodeoxyglucose positron emission tomography and its prognostic value in lung cancer. AB - OBJECTIVE: Positron emission tomography (PET) is being increasingly used as an accurate and non-invasive modality in diagnosis, staging and post-therapy assessment in patients with lung cancer. In this study, we examine whether the uptake of [(18)F]fluorodeoxyglucose (FDG), a marker of increased glucose metabolism in neoplastic cells, is of prognostic value in patients with primary lung cancer. METHODS: We have retrospectively analyzed 77 patients (mean age, 63. 0 years; male/female ratio, 53:24) with primary lung cancers who underwent whole body and localized thoracic PET as part of their diagnostic and staging procedures prior to consideration of surgical resection. The standardized uptake value (SUV) of injected FDG for each primary lesion was correlated with tumour histology and the patient's clinical outcome. RESULTS: A SUV of 20 or greater was found to be of significant prognostic value. The chance of survival (with 95% confidence intervals (CI)) at 12 months post-surgery for the various SUV groups was as follows: 75.2% (59.6-85.5) for SUV<10; 67.5% (29.0-88.2) for SUV 10-<12; 63.6% (29.7-84.5) for SUV 12-<15; 66.7% (19.5-90.4) for SUV 15-<20; 16.7% (0.01 0.52) for SUV>20. A SUV of 20 or more is associated with a 4.66 times increase in hazard, compared with lower levels of SUV. We found no significant correlation between tumour histology and SUV. CONCLUSION: We have previously reported on the significant advantages of PET in the staging and surgical care of patients with lung cancer. The present study adds further support for an additional prognostic role for PET in the management of thoracic malignancy as determined by the amount of labelled-FDG taken up by the primary lesion. PMID- 11024378 TI - Radiotherapy versus follow-up in the treatment of pathological stage Ia and Ib non-small cell lung cancer. Early stopped analysis of a randomized controlled study. AB - OBJECTIVE: This is an analysis of a randomized controlled clinical trial planned to evaluate the effects of adjuvant radiotherapy (AR) on the local recurrence rate in patients with non-small cell lung cancer (NSCLC) with pathological stage (pStage) Ia (pT1N0) and Ib (pT2N0). The effects of AR on the long-term survival have also been marginally evaluated. MATERIALS AND METHODS: This clinical trial was planned with the hypothesis that AR on pStage Ia and Ib, R0 NSCLCs was effective on local recurrence rate. From July 1989 through March 1997, 104 patients with NSCLC who presented with pStage Ia and Ib have been observed and treated and entered the study. Male/female ratio was 91:13; the mean age was 62 years (range 41-75 years). All patients underwent major pulmonary resection and homolateral standard hilar and mediastinal lymph node dissection. pStage was T1N0 in 29 and T2N0 in 75 cases. Patients have been randomized 'by chance' into two groups (G1 and G2). G1 received radiotherapy, G2 did not receive any adjuvant treatment. Fifty-two patients entered G1 and 52 entered G2. RESULTS: Post operative mortality was nil. Seven patients have been excluded from the study (four in G1 and three in G2), due to incomplete follow-up data. We do not report any radiotherapy-related complication or deterioration of lung function. The treatment effect on the local recurrence rate demonstrated a clearly significant protective effect of the AR. No statistically significant difference was found from the comparison of the 5-year survival rate of the treated (83%) versus untreated (70%) patients. No detrimental effect of the radiotherapy has been assessed. CONCLUSIONS: AR in the treatment of pStage Ia and Ib NSCLC has been well tolerated and had a significant relative effect on the local recurrence rate but did not significantly modify overall survival even if a positive trend in the group of treated patients is reported. PMID- 11024380 TI - Pneumonectomy for inflammatory lung disease. AB - OBJECTIVE: Recent surgical literature has highlighted the dangers of pneumonectomy for inflammatory lung disease; therefore the assessment of the risk/benefit ratio of our departmental policy. METHODS: Patients undergoing pneumonectomy for inflammatory lung disease during two 2-year periods, 1991-1992 and 1996-1997 inclusive, were retrospectively analyzed. Clinical indications for investigation and surgery, and radiographic findings were determined. Some comparisons between the two periods were drawn. Rates of morbidity and mortality were the principle outcome measures. RESULTS: One hundred and fifty-five patients, 116 males, 39 females, with an average age of 30.2 years ranging from 1 68 years, underwent pneumonectomy for ongoing features of productive cough, haemoptysis (two emergencies) and chronic empyema all with either bronchographic or computed tomography (CT) evidence of destroyed lung. One hundred and fourteen (72%) had or had had tuberculosis at time of surgery. Histology showed bronchiectasis in 53 (34%), end-stage disease in 49 (31.6%) and active tuberculosis in 48 (30.9%). Over 90% of the patients were free of disease at discharge. Mortality was two (1.2%). Morbidity (23%) included post-pneumonectomy empyema 23 (14.8%), bleeding three (1.9%), broncho-pleural fistula three (1.9%), with wound sepsis in one (0.6%) and thoracic duct injury in one (0.6%). Three groups were identified, (1) pneumonectomy through empyema - a risk group, (2) pneumonectomy in active tuberculosis and (3) pneumonectomy in children. Twenty three post-pneumonectomy empyemas (PPE) occurred with 21 of these following pneumonectomies through empyema (PTE), six PPEs followed 27 PTEs for active tuberculosis. Fourteen of the 21 empyemas following pneumonectomy through empyema were initially sterilized. Finally 15/23 (65%) of all PPEs were sterilized. Pneumonectomy in active tuberculosis did not carry the mortality or morbidity experienced by others. Pneumonectomy in children was remarkably uncomplicated, with one PPE occurring. CONCLUSIONS: This ongoing study shows pneumonectomy for inflammatory lung to be safe, with good results. Tuberculosis, being so common, adequate pre-operative and operative cover with anti-tuberculosis drugs may enhance results. PMID- 11024381 TI - A comparative evaluation of intrathoracic latissimus dorsi and serratus anterior muscle transposition. AB - BACKGROUND: Comparison of intrathoracic latissimus dorsi (LD) versus serratus anterior (SA) muscle transposition for treatment of infected spaces, broncho pleural fistulae, and for prophylactic reinforcement of the mediastinum after extended resections following induction therapy. PATIENTS AND METHODS: Twenty LD and 17 SA transfers were performed for prophylactic reinforcement (11 LD; nine SA), and treatment of infections (nine LD; eight SA) from 1995 to 1998. RESULTS: The 30-day mortality was 0% following prophylactic reinforcement and 29% following treatment of infections (three LD; two SA). Prophylactic mediastinal reinforcement was successful in 11 of 11 patients with LD and nine of nine with SA transpositions, and treatment of infected spaces in eight of nine patients with LD and two of three with SA transfers. Morbidity requiring re-intervention consisted of flap necrosis (one LD), bleeding (one SA), and skin necrosis over a winged scapula (one SA). Subcutaneous seromas and chest wall complaints were more frequent following LD (45 and 36%, respectively) compared with SA transfers (29 and 27%, respectively), whereas impaired shoulder girdle function was more frequent after SA than after LD transfer (27 vs. 21%). CONCLUSION: Intrathoracic LD and SA muscle transpositions are both efficient for the prevention or control of infections following complex thoracic surgery, and are both associated with similar and acceptable morbidity and long-term sequelae. PMID- 11024382 TI - Evolution of antimicrobial prophylaxis in cardiovascular surgery. AB - OBJECTIVE: To examine the optimal duration of antibiotic prophylaxis in major cardiovascular surgery. MTHODSs: In the past 15 years, four prospective randomized, controlled studies, conducted by the same group of authors, compared seven prophylactic antimicrobial regimens in 2970 patients undergoing major cardiovascular surgery. In 1980/81, a 4-day cefazolin (CFZ) prophylaxis was compared with a 2-day cefuroxime (CFX) administration (n=566). In 1982/83, a 2 day CFX prophylaxis was compared with a two shot ceftriaxone (CRO) prophylaxis (n=512). In 1984/87, a 1-day CFZ prophylaxis was compared with a single shot prophylaxis of CRO (n=883). In 1994/1995, a 4 day combination of amoxicillin (AM) and netilmicin (NET) prophylaxis was compared with a single shot prophylaxis of CFX (n=1009). RESULTS: Total infection rate varied between 4.5 and 5.7%, despite different antimicrobial regimen used and their varying duration. Wound infection rate was 1.1% (range 0.4-2.5%), sepsis rate was 0.8% (range 0.4-1.6%), pneumonia rate 2% (0.7-2.9%), urinary tract infection rate 0.4% (range 0-1.4%), and central venous catheter-related infection rate was 0.4% (0-1%). The 30-day mortality rate was 1.3% (range 0.4-2%). All these differences were not statistically significant. CONCLUSIONS: A low infection rate (range 4.5-5.7%) occurred despite changes in duration of various prophylactic antibiotic regimen with cephalosporins of first, second or third generation. As a single shot prophylaxis could nowadays successfully be used in cardiovascular surgery, no postoperative antibiotics should be used, unless an intraoperative or a postoperative infection is documented or in presence of major perioperative complications. PMID- 11024383 TI - Ten years experience with lung and heart-lung transplantation in primary and secondary pulmonary hypertension. AB - OBJECTIVE: Patients with primary pulmonary hypertension (PPHT) have a worse natural outcome compared with those with secondary pulmonary hypertension in Eisenmenger's syndrome (ES) and chronic pulmonary embolism (CPE). Lung transplantation (SLTx, DLTx, HLTx) still remains the only therapeutical option for patients with this type of endstage lung disease. METHODS: From 1988 to 1998, 63 patients underwent lung transplantation for PPHT (n=29, 9 m, 20 f, 2 SLTx, 14 DLTx, 13 HLTx), ES (n=29, 13 m, 16 f, 2 SLTx, 3 DLTx, 24 HLTx) or CPE (n=5, 2 m, 3 f, 1 SLTx, 2 DLTx, 2 HLTx). Groups were comparable for NYHA functional class, preoperative pulmonary arterial pressure, recipient and donor age, ischemic time, necessity and duration of cardiopulmonary bypass and cross-match. RESULTS: The 1 , 3- and 5-year survival was 52, 40 and 35% for the PPHT-group, 83, 78 and 74% for the ES-group and 80, 60 and 60% for the CPE-group, respectively (P=0.026, P=0.033, P=0.082 for 1-, 3- and 5-year survival). Patients following DLTx showed a lower 1-year survival rate as compared with patients after HLTx both in PPHT patients (36 vs. 62%, P=0.091) and in ES patients (67 vs. 83%, P=0.213). The incidence of bronchiolitis obliterans syndrome was 29% at 1 year and 45% at 3 years for the PPHT-group vs. 17 and 65% for the ES-group (n. s. in between groups). Excluding postoperative ventilation time (PPHT-group: 26.8+/-24.0 days vs. ES-group: 16.1+/-30.8 days, P=0. 011) and a higher incidence of infectious causes of death (PPHT-group n=8 vs. ES-group n=1, P=0.017) groups were comparable with regard to their postoperative courses. CONCLUSIONS: It is concluded, that predominantly the underlying primary disease influences graft survival after lung transplantation in patients with pulmonary hypertension compared with all other patient and procedure dependent factors. Lung transplantation in patients with PPHT requires further investigations to achieve results comparable with other indications. PMID- 11024384 TI - Mid-term results of the valve-on-valve technique for bioprosthetic failure. AB - OBJECTIVE: Redo operations for bioprosthesis malfunction can sometimes be technically very demanding and cardiac structures may be damaged. Excising only the leaflets of the damaged bioprosthesis and leaving the old ring in situ on which the 'new' mechanical valve is sutured can, in very selected cases, represent a solution. METHODS: Twenty-two patients were operated on, with the valve-on-valve technique, from September 1991 through December 1992. There were three operative deaths. RESULTS: The surviving 19 patients were followed-up from 83 to 98 months (mean 90.5 months.). There were two late deaths. The patients were examined clinically and with transthoracic and transesophageal echocardiograms. All patients were in good condition and the echocardiographic examinations showed no clinically important gradients across the prostheses. CONCLUSIONS: The valve-on-valve technique, in certain difficult situations, can give successful mid-term results. PMID- 11024385 TI - Left ventricular regional wall motion, ejection fraction, and geometry after partial left ventriculectomy. Influence of associated mitral valve repair. AB - OBJECTIVE: Left partial ventriculectomy has been proposed for treatment of heart failure. We investigated the effects of isolated left partial ventriculectomy and left partial ventriculectomy associated with mitral annuloplasty on refractory heart failure due to idiopathic dilated cardiomyopathy. METHODS: Nineteen patients underwent partial left partial ventriculectomy associated with mitral annuloplasty and six patients isolated left partial ventriculectomy. In two patients the left partial ventriculectomy associated with mitral annuloplasty was combined with tricuspid annuloplasty. We evaluated before and after the surgery (24+/-14 days): the functional class, left ventricular ejection fraction (LVEF), right ventricular ejection fraction (EF), regional wall motion, hemodynamics, mitral regurgitation, left ventricular geometry and coronary angiography. RESULTS: For the overall group LVEF improved from 14.5+/-8.0 to 30.3+/-12.2% (P<0.0002) and right ventricular EF from 21.2+/-7.1 to 28.4+/-8.3% (P<0.002). In patients who underwent left partial ventriculectomy associated with mitral annuloplasty LVEF increased from 14.5+/-8.6 to 29.5+/-12.2% (P<0. 002). Isolated left partial ventriculectomy increased LVEF from 13. 5+/-7.5 to 31.5+/-11.1% (P<0.04). Distal segments of marginal branches of the circumflex artery were not visualized by coronary angiography. Left partial ventriculectomy associated with mitral annuloplasty reduced the wedge pressure from 25.0+/-12.1 to 18.0+/-7. 0 mmHg (P<0.03) and increased cardiac output from 3.8+/-0.8 to 4. 6+/-1.1 l/min (P<0.004), while isolated left partial ventriculectomy increased cardiac output from 3.7+/-1.0 to 4.8+/-1.3 l/min (P<0.03). Regional wall motion increment was more evident in anterolateral region from 4.2+/-6.8 to 14+/-8.3% (P<0.002) except in two patients. Left ventricular geometry changed in most patients, but a homogeneous pattern was not identified. Seven patients died during a mean follow up of 546+/-276 days. Survivors had improvement in functional class. Augmentation of LVEF >5% was associated with a favorable clinical outcome with improvement in clinical status without death. CONCLUSIONS: Effects of left partial ventriculectomy are not necessarily dependent upon reduction of mitral regurgitation or changes in left ventricular geometry. However, risk of death after the surgery must be reduced for a clinical application. PMID- 11024386 TI - Circulating donor-specific cytotoxic T lymphocytes with high avidity for donor human leukocyte antigens in pediatric and adult cardiac allograft valved conduit recipients. AB - OBJECTIVE: Specific immunological responses may be involved in the process of cryopreserved allograft valved conduit (AVC) degeneration, which is more frequently seen in young recipients. Rejection of heart and corneal allografts is preceded by an increase in the fraction of cytotoxic T lymphocytes (CTL) with high avidity for donor human leukocyte antigens (HLA) circulating in both peripheral blood and the affected graft. These donor-specific high-avidity CTLs are regarded as the destructive cells capable of causing graft damage. To monitor the precursors of these cells (CTLp) in young and adult AVC recipients, in vitro quantitative tests were performed on sequentially taken blood samples to quantitate CTLp frequencies and their avidity for donor antigens. METHOD: Six children and nine adults who received a cryopreserved AVC in the period between 1994 and 1997 were included in the study. From these patients, two to six blood samples were obtained up to 3 years after valve implantation. The number of circulating CTLp present within the peripheral blood mononuclear cell (PBMC) population was determined by limiting dilution analysis (LDA). The fraction of CTLp with high avidity for donor HLA class I was determined by addition of CD8 monoclonal antibodies (mAb) during the cytotoxic phase of the assay. Third-party stimulator cells were used to verify the donor-specificity of the response. RESULTS: The number of donor-specific CTLp increased significantly in the period 6-12 months after AVC implantation, while third-party-specific CTLp frequencies were not affected. Additionally, we found a significant increase of the high avidity fraction of CTLp directed against donor antigens as early as during the first 6 months after AVC implantation. The fraction of high-avidity CTLp remained significantly higher post- compared with pre-implantation, even after 12 months. We observed no significant difference in the kinetics of CTLp frequencies between pediatric and adult AVC recipients. CONCLUSION: Implantation of cryopreserved human AVC induces an increase in the total number of circulating CTLp directed against donor HLA class I in both adults and children. The shift towards more destructive high-avidity CTLp in the peripheral blood indicates their potential damaging effect towards the heart valve allograft. PMID- 11024387 TI - Results of primary two-patch repair of complete atrioventricular septal defect. AB - OBJECTIVE: The policy of primary repair of complete atrioventricular septal defect (CAVSD), using a two-patch technique, was evaluated with special attention to the risk of implantation of a prosthetic atrioventricular (AV) valve. METHODS: From 1986 to 1999, all 97 patients who underwent primary repair for CAVSD were included in a retrospective analysis. Seventy-five patients (75%) had Down's syndrome. Preoperative echocardiographic AV valve regurgitation was absent or limited in 85 (88%), moderate in seven (7%) and severe in five (5%). Fifty-six patients (58%) were on diuretics, six (6%) on artificial ventilation and four (4%) were on inotropic support. The mean age at operation was 10.2 months (SD, 16.4), with a mean weight of 5.9 kg (SD, 3.7). RESULTS: Early mortality comprised three patients (4%), and late mortality two patients. Follow up was complete and comprised 402 patient-years (mean, 4.5 years; SD, 3.2). The cumulative survival at 10 years was 93% (95% CI, 89-97%). Multivariate analysis with regard to mortality revealed no associations with any of the analyzed factors. Eight patients were reoperated, all for regurgitant left AV valve. The reoperation-free survival at 10 years was 83% (95% CI, 75-91%). Multivariate analysis with regard to reoperation showed being on preoperative diuretics to be a decreasing risk factor (Odd's Ratio (OR), 0.13; 95% CI, 0.00-0. 99; P=0.005) and significant postoperative left AV valve regurgitation to be an increasing risk factor (OR, 9.90; 95% CI, 1. 90-53.0; P=0.001). Only one prosthetic valve was implanted (annual linearized risk of 0.002/patient-year). At the latest follow up of the surviving patients, left AV valve regurgitation was absent or limited in 83 (90%) and moderate in nine (10%). Right AV valve regurgitation was absent or limited in all 92 (100%) patients. All surviving patients are thriving well, seven (8%) of whom are on diuretics. CONCLUSIONS: Primary repair of CAVSD with a two-patch technique, including cleft closure of the left AV valve, has good clinical and functional results without problems for the right-sided AV valve. The need for prosthetic valve implantation for the left AV valve is minimal. PMID- 11024388 TI - The management of pulmonary atresia, ventricular septal defect, and multiple aorta pulmonary collateral arteries by definitive single stage repair in early infancy. AB - OBJECTIVE: The management of infants and children with pulmonary atresia, ventricular septal defect, and multiple aorta pulmonary collateral arteries (PA/VSD/MAPCA) has proven to be challenging. Therapeutic approaches have included staged unifocalization, shunting, coiling of collateral vessels, and heart/lung transplantation. Results have been variable and frustrating. Hoping to take advantage of growth potential in pulmonary segments supplied by MAPCA, a more radical approach was adopted in March of 1997. This consists of single stage complete unifocalization with closure of the ventricular septal defect and establishment of right ventricular to pulmonary arterial continuity with a cryopreserved pulmonary allograft (Rastelli type correction) through a midline sternal incision. METHODS: During an 18-month period, eleven consecutive infants with PA/VSD/MAPCA underwent complete surgical correction. The ages ranged from 5 days to 5 months. Weights ranged from 2.2 to 5.6 kg. Through a standard median sternotomy incision, the pericardium and both pleural spaces were opened. Normothermic cardiopulmonary bypass was instituted. Section of all collaterals was accomplished without hypoxemia, and all collaterals were ligated at their origin from the aorta. They were then brought through posterior mediastinum to construct a pulmonary artery confluence. The ventricular septal defect was closed, and continuity was established between the right ventricle and the newly created pulmonary artery confluence with cryopreserved allografts. RESULTS: Ten of 11 patients survived operation, with postoperative courses that were uncomplicated. Length of stay ranged from 7-16 days, with a median length of stay of 11 days. One perioperative death occurred in a patient with preoperative co morbidities of necrotizing enterocolitis, with no functioning gastrointestinal tract, intraventricular hemorrhage, and ventilator dependency since birth. At angiography, this patient has no demonstrable central pulmonary arteries and multiple diminutive aorta pulmonary collaterals. Autopsy revealed no demonstrable pulmonary arteries within the pulmonary parenchyma. All patients have been followed closely, and have grown normally. Two patients undergone repeat cardiac catheterization because of the echocardiographic demonstration of right ventricle pressures that had exceeded 50% of systemic. Both patients were treated with balloon angioplasty and one of these patients has had stenting of stenotic pulmonary arterial segments. No other patients have required additional hospitalization. Right ventricular pressures have remained less than fifty percent of systemic by echocardiographic assessment in all other patients. CONCLUSIONS: We feel that a single stage correction of pulmonary atresia, ventricular septal defect, and multiple aorta pulmonary collateral arteries can be accomplished in early infancy with acceptable morbidity and mortality. The initiation of normothermic cardio-pulmonary bypass greatly facilitates dissection of collaterals and prevents hypoxemia. Interventional cardiology with balloon angioplasty and stenting of abnormal pulmonary arterial segments is both an important essential adjunct in the management of these patients. We remain optimistic that a single stage approach to this complex lesion coupled with cardiac catheterization, balloon angioplasty, and stenting, will provide long term results superior to staged approaches. PMID- 11024389 TI - Surgical treatment of Wolf-Parkinson-White syndrome during plastic operations in patients with Ebstein's anomaly. AB - OBJECTIVE: Ebstein's anomaly is the most common pathology associated with the accessory conduction pathways. METHODS: From January 1990 to August 1999 48 patients underwent surgical repair of Ebstein's anomaly by various plastic techniques. The pathways were identified and characterized at preoperative electrophysiologic mapping in 17 (34.5%) patients. There were seven males and ten females. The patients age ranged 6-35 years (mean 12.7+/-2.1 years). Five patients were in NYHA class II and were in 12 in NYHA class III. The cardiothoracic ratio ranged from 0.59 to 0.69 (mean 0.65+/-0.08). Tachycardia was present in 15 patients with Wolf-Parkinson-White (WPW) syndrome. Atrial septal defect was present in 12 patients (70. 6%). Accessory conduction pathways were in the right posterior septal area in seven patients, in right posterior septal area and free wall in seven patients and in right free wall in three. The pathways were successfully ablated in all patients during cardiopulmonary bypass. RESULTS: There were no deaths, no relapses. Follow-up ranged from 4 months to 7 years (mean 4.7+/-1.1 years). There were no late deaths, but one patient required successful tricuspid valve replacement because of severe tricuspid insufficiency. At follow-up 57.8% of patients were in NYHA class I. CONCLUSIONS: Preoperative electrophysiologic study allows to identify the accessory conduction pathways in patients with Ebstein's anomaly. The combined approach of tricuspid valve repair and surgical ablation of accessory atrioventricular connections has been proved safe and effective. This operative procedure allows to improve functional results in patients with Ebstein's malformation and concomitant accessory conduction pathways. PMID- 11024390 TI - A case of multiple synchronous localized fibrous tumor of the pleura. AB - We report a patient with two synchronous distinct masses in the same hemithorax both of which got the diagnosis of benign localized fibrous tumor of the pleura. The plain chest X-ray was rather obscured due to a large left-sided pleural effusion, but her subsequent computerized chest tomography revealed a heterogeneous hypodense soft tissue mass, which was pleural in origin, sitting on the diaphragm bathed in fluid. At thoracotomy, we detected two distinct masses in the left hemithorax, both arising from the visceral pleura via their vascular pedicles. PMID- 11024391 TI - Scapular osteochondroma with reactive bursitis presenting as a chest wall tumour. AB - A 32-year-old male presented with a painful, rapidly enlarging chest wall mass. A malignant chest wall neoplasm was suspected. A CT scan was performed which showed a mass extending from under the scapular and an exostosis arising from the anterior surface of the scapular. The mass and exostosis were resected resulting in complete resolution of symptoms. Histological examination showed the mass to be a reactive bursa, with no evidence of neoplasia. PMID- 11024392 TI - Constrictive calcified pericarditis associated with non-treated pulmonary tuberculosis. PMID- 11024394 TI - Association between new electrocardiographic abnormalities after coronary revascularization and five-year cardiac mortality in BARI randomized and registry patients. AB - There are few data comparing the relative frequency of new electrocardiographic (ECG) abnormalities after coronary artery bypass grafting (CABG) compared with percutaneous transluminal coronary angioplasty (PTCA) and their association with long-term cardiac mortality. The study population consisted of 3,373 patients who were either randomized or eligible to be randomized to CABG or PTCA in the BARI trial. The frequency of new postprocedural ECG abnormalities was significantly greater after a CABG procedure than after PTCA. The incidence of new postprocedural major Q waves, ST-segment elevation, and T-wave abnormalities were significantly more frequent after CABG. After PTCA (n = 1,869), the 5-year cardiac mortality rates associated with the new development of major Q waves, ST segment elevation, ST-segment depression, T-wave abnormalities, or no abnormality was 18.1%, 8.5%, 8.9%, 6.0%, and 5.4%, respectively. After CABG (n = 1,427), 5 year cardiac mortality rates were 8.0%, 4.2%, 3.8%, 2.8%, and 3.7%, respectively. The adjusted relative risk of 5-year cardiac mortality for new Q-wave abnormalities was 2.6 after CABG (p <0.04) and 4.6 after PTCA (p <0.01). Thus, patients who undergo CABG have more postinitial procedural ECG abnormalities than patients who undergo PTCA. Cardiac mortality is significantly increased by the new development of postprocedural Minnesota code Q-wave abnormalities regardless of whether patients undergo CABG or PTCA. PMID- 11024393 TI - Effect of smoking on lipid and thrombogenic factors two months after acute myocardial infarction. AB - Cigarette smoking is linked to increased cardiac morbidity and mortality, and has been shown to affect both lipid profiles and thrombotic factors in healthy subjects. However, the influence of smoking on the atherothrombotic environment has not been studied in a large population of patients after acute myocardial infarction (AMI). Blood samples and medical history, including smoking status, were obtained from 1,045 patients at a 2-month visit after AMI. Smokers were asked to refrain 24 hours before the visit, but not all complied. Measurements included total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, apolipoprotein-B, apolipoprotein-A, triglycerides, factor VII, factor VIIa, von Willebrand factor, D-dimer, and plasminogen activator inhibitor. There were 247 current, 443 past, and 349 nonsmokers. After adjustment for clinical variables, current smokers had higher levels of total cholesterol and apolipoprotein-B than past and nonsmokers (p <0.01). High-density lipoprotein cholesterol and apolipoprotein-A levels were similar between groups. Fibrinogen was elevated in current (p = 0.001) and past (p = 0.029) smokers, compared with nonsmokers. Smokers who smoked within 24 hours of blood sampling had higher apolipoprotein-B (p = 0.005), total cholesterol (p = 0.001), and fibrinogen (p = 0.015) levels than those who refrained from smoking. In conclusion, postinfarction patients, who historically have higher levels of atherogenic lipids than healthy subjects, have increased levels of these lipids attributed to active smoking. After smoking cessation, lipid profiles approach nonsmoker levels, but fibrinogen remains elevated. Smoking within 24 hours of blood sampling was associated with further adverse prothrombotic and lipogenic effects. PMID- 11024395 TI - Safety and efficacy of an accelerated dobutamine stress echocardiography protocol in the evaluation of coronary artery disease. AB - Although dobutamine requires up to 10 minutes to achieve steady state, dobutamine stress echocardiography is routinely performed using stepwise increments at 3 minute intervals. Consequently, the full effect of any infusion rate is not attained before the dobutamine dose is advanced to the next level. This study sought to test the safety and efficiency of high-dose continuous dobutamine infusion. One hundred consecutive patients underwent an accelerated protocol using a constant infusion of 50 microg/kg/min. In the absence of a stress echocardiographic end point (>/=85% of maximal predicted heart rate, new wall motion abnormalities, hypotension, arrhythmia, or intolerable symptoms), dobutamine infusion was discontinued at 10 minutes. Hemodynamic responses and adverse effect profile were compared with 100 patients who underwent a standard stepwise dobutamine stress protocol. Peak heart rate (140 +/- 16 vs 140 +/- 19 beats/min, p = 0.95) and systolic blood pressure (169 +/- 32 vs 162 +/- 31 mm Hg, p = 0.08) were similar in both protocols. Accelerated dobutamine administration produced a rapid increase in heart rate (12.5 +/- 6.2 vs 5.7 +/- 2.6 beats/min, p <0.001), and a substantial reduction in test duration (6.4 +/- 2.4 vs 12.9 +/- 3.0 minutes, p <0.001). The mean weight-adjusted cumulative dobutamine dose was lower in the accelerated protocol group (320 +/- 111 vs 353 +/- 133 microg/kg, p = 0.016). No significant differences were noted between the 2 groups with respect to various side effects. These data demonstrate that a high-dose, single-stage dobutamine echocardiographic stress protocol is a feasible, well-tolerated alternative to standard dobutamine stress echocardiography, and results in a substantial reduction in test time while maintaining a low complication rate. PMID- 11024396 TI - Clinical and electrocardiographic variables associated with increased risk of ventricular septal defect in acute anterior myocardial infarction. AB - It is unknown whether the risk factors associated with the development of ventricular septal defect (VSD) after acute myocardial infarction (MI) remain the same when thrombolytic therapy is used, nor have specific electrocardiographic patterns of acute MI associated with the development of VSD been identified. Our study population included patients with an anterior MI enrolled in the GUSTO-I study. Baseline clinical data were collected prospectively for all patients. Patients in whom VSD was suspected by the local investigators at each site were evaluated retrospectively. Baseline clinical and electrocardiographic variables were compared between 2 groups: 10,847 patients without VSD (99.6%) and 48 patients with confirmed VSD (0.4%). Multivariate analysis showed the following clinical variables to be independent predictors of VSD: age (odds ratio [OR] 2.19, 95% confidence intervals [CI] 1.62 to 2.98; p <0.001), female gender (OR 5.07, 95% CI 2.70 to 9.98; p <0.001), and lack of previous angina (OR 2.11, 95% CI 1.12 to 4.29; p = 0.021). Two electrocardiographic variables predicted acute VSD: the magnitude of ST deviation in lead III (OR 1.55, 95% CI 1.12 to 2.21; p = 0.007) and in lead V(2) (p <0.001). However, the relation between the ST amplitude in lead V(2) and the risk for VSD was nonlinear. In patients with anterior MI who underwent thrombolysis, the risk factors for VSD were age, female gender, and lack of previous angina. Previous infarction was not a risk factor. Less ST-segment depression in lead III was a predictor of VSD. PMID- 11024397 TI - Comparison of platelet activation in unstable and stable angina pectoris and correlation with coronary angiographic findings. AB - We sought to investigate the relation between platelet activation and the angiographic evidence of ruptured plaque in patients presenting with unstable and stable angina pectoris. We prospectively enrolled 25 consecutive patients (5 women and 20 men, mean age 62 +/- 3 years), 17 with unstable angina and 8 with stable angina. Systemic venous blood samples were collected within 4 to 6 hours of admission for flow cytometry analysis. Activation-dependent epitope CD63 and glycoprotein IIb/IIIa on the platelet membrane were assayed. Fibrinogen levels were also measured. All patients with unstable angina underwent cardiac catheterization and had angiographic evidence of ruptured plaque. Of the patients with stable angina, 5 underwent coronary angiography with smooth noncomplex lesions and 3 had negative technetium-99m sestamibi stress tests. Patients with unstable angina were characterized by 39% higher levels of fibrinogen than patients with stable angina (423 +/- 304 vs 304 +/- 51 mg/dl, p = 0.004). The percentage of platelets positive for the activation-dependent epitope CD63 was 5 times higher in patients with unstable than stable angina (14.6 +/- 5.6% vs 2.75 +/- 1.6%, p = 0.0026). They also had a 15% higher expression of their glycoprotein IIb/IIIa (517 +/- 79 vs 449 +/- 50 mean fluorescence intensity, p = 0.038). Thus, this study establishes a direct relation between the morphology of ruptured plaque and platelet activation in patients with unstable angina. This may allow for further risk stratification. Patients with unstable complex lesions had a fivefold higher expression of the platelet activation epitope CD63 than patients with stable angina. Furthermore, they had 15% more glycoprotein IIb/IIIa aggregation sites expressed on their platelet membrane, thus indicating an intense thrombogenic potential. PMID- 11024398 TI - Effect of common methylenetetrahydrofolate reductase gene mutation on coronary artery disease in familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is an autosomal dominant disorder characterized by primary hypercholesterolemia and premature coronary artery disease (CAD). However, the development of CAD in FH shows considerable interindividual variations. Elevated levels of plasma homocysteine have been recognized as independent risk factors for CAD. A 5,10-methylenetetrahydrofolate reductase (MTHFR) gene mutation (valine [V] was substituted for alanine [A]) has been reported to be associated with elevated levels of plasma homocysteine in mutant homozygotes (i.e., VV). We studied 199 consecutive male heterozygous FH patients, 99 with and 100 without CAD. In the CAD group, genotype VV and V alleles were significantly more frequent than in the non-CAD group (15% vs 7% in genotypes [p = 0.035] and 0.41 vs 0.30 in alleles [p = 0.017]). The mean ages at onset in the CAD group were 50, 51, and 43 years for genotypes AA, AV, and VV, respectively (p <0.05); the age of onset of CAD in genotype VV was significantly lower than in the other 2 genotypes. Kaplan-Meier survivor curves indicated that the development of CAD was significantly accelerated by MTHFR mutation, probably in a gene dose-dependent manner. Furthermore, only MTHFR genotype VV was shown to be an independent predictor of the early onset of CAD in a stepwise multiple regression analysis. The mean plasma homocysteine levels of genotype VV were significantly higher than those of the other 2 genotypes. Thus, the MTHFR mutation appears to accelerate the onset of CAD through elevation of plasma homocysteine levels in male heterozygous patients with FH. PMID- 11024399 TI - Location of death (in-hospital or out-of-hospital) and type of death (arrhythmic, nonarrhythmic, noncardiac) after inducible sustained ventricular tachyarrhythmias after syncope, sustained ventricular tachycardia, or nonfatal cardiac arrest (the ESVEM trial). AB - The ESVEM Trial evaluated methods to guide antiarrhythmic drug use in patients with spontaneous, inducible sustained tachyarrhythmias at electrophysiologic testing and frequent ventricular premature complexes (VPCs) per hour (>/=10). We assessed the relation between location (in-hospital or out-of-hospital) and classification of death (arrhythmic, nonarrhythmic, cardiac and/or noncardiac) for 486 randomized patients. Deaths were classified as out-of-hospital arrhythmic deaths if arrhythmic death occurred out-of-hospital, or if an arrhythmia preceded hospital admission and directly caused death. Of the 486 randomized patients, 188 (39%) died during 6 years of follow-up. The location and type of death could be determined clearly in 171 patients (91%). Ninety-one deaths were in-hospital (53%); 80 were out-of-hospital (47%). Arrhythmic deaths occurred in 85% out-of hospital patients and in 30% in-hospital patients (p <0.001). Baseline characteristics were comparable for patients with out-of-hospital and in-hospital arrhythmic deaths. Twenty-seven of 95 arrhythmic deaths occurred in-hospital (28%); 72% occurred out-of-hospital. Out-of-hospital arrhythmic death accounted for 40% of deaths for which location and type of information were available. The 1- and 4-year actuarial out-of-hospital arrhythmic death rates were 9% and 18%, respectively. Of nonarrhythmic cardiac deaths, 91% were in-hospital and 9% were out-of-hospital. Of noncardiac deaths, 74% were in-hospital and 26% were out-of hospital. Similar results were seen in the 296 patients for whom a drug was considered to be effective. Thus, over half the deaths in the ESVEM trial occurred in-hospital. The long-term actuarial risk of out-of-hospital arrhythmic death in ESVEM was unexpectedly low. PMID- 11024400 TI - Risk factors for thromboembolism in patients with paroxysmal atrial fibrillation. AB - There is some controversy concerning which clinical characteristics predict thromboembolism and whether treatment with class I antiarrhythmic drugs reduces thromboembolim in patients with paroxysmal atrial fibrillation (AF). This retrospective, multicenter study was undertaken to determine risk factor or factors for thromboembolism in patients with paroxysmal AF. Seven hundred forty patients with paroxysmal AF (mean age 56 years) without prior thromboembolic events were followed retrospectively. Cerebral thromboembolism, including transient ischemic attack and embolism of peripheral arteries, were selected as primary end points. Independent risk factors were determined with multivariate analysis, and event-free survival curves were estimated. During 3.4-year follow up period, primary end points occurred in 55 patients (2.2% per year). Patients with thromboembolism had a higher prevalence of underlying heart disease (p <0.01), less frequent treatment with antiarrhythmic drugs (p <0.01), and received diuretics more often (p <0.01) compared with patients without thromboembolism. Age (>/=65 years, RR 3.33, p = 0.0001) and gender (male, RR = 2, p = 0.0291) emerged as predictors of thromboembolism by multivariate analysis with Cox's proportional hazard model. Treatment with antiarrhythmic drugs (RR = 0.57, p = 0.0578) and aspirin (RR = 0.52, p = 0.1094) showed trends toward reducing thromboembolic risks. It is suggested that elderly men (>/=65 years) with paroxysmal AF are at risk for thromboembolism, but the risk tended to be reduced by treatment with antiarrhythmic drugs and aspirin. PMID- 11024401 TI - Comparison of left ventricular outflow tract obstruction in interruption of the aortic arch and in coarctation of the aorta, with diagnostic, developmental, and surgical implications. AB - A morphometric comparison of the anatomic causes of left ventricular (LV) outflow obstruction in interruption of the aortic arch and in coarctation of the aorta with ventricular septal defect (VSD), based on 30 postmortem cases of each, revealed that posterior malalignment of the conal septum with a conoventricular VSD was significantly more prevalent with interruption (93%) than with coarctation (47%) (p <0.001). The ratio of the aortic valve diameter-to-the pulmonary valve diameter, which provided a quantitative index of the degree of posterior conal septal malalignment and of the consequent LV outflow tract obstruction at and immediately below the level of the aortic valve, was significantly smaller with interruption (40%, who underwent cardiac catheterization between January 1984 and December 1996 at Duke University Medical Center. The median age for the entire cohort was 63 years; 25% of the population was >71 years old. In addition, 55% of the patients were women, 65% had ischemic heart disease, 28% had a history of diabetes, and 62% had a history of hypertension. The median ejection fraction was 58%. One third of the patients had multivessel disease by coronary angiography. The overall 5-year mortality of the total population was 28%. The independent predictors of mortality (p <0.05) using a multivariable Cox proportional hazard model were age, class IV symptoms, ejection fraction, coronary artery disease index, diabetes, peripheral vascular disease, and minority ethnic group. Heart failure with preserved systolic function is characterized by unique clinical and angiographic characteristics associated with a 5-year mortality rate of 28%. Furthermore, several clinical and angiographic characteristics are predictive of long-term survival. PMID- 11024403 TI - Evolution of aortic dissection after surgical repair. AB - Patients after aortic dissection repair still have long-term unfavorable prognosis and need careful monitoring. The purpose of this study was to analyze the evolution of aortic dissection after surgical repair in correlation to anatomic changes emerging from systematic magnetic resonance imaging (MRI) follow up. Between January 1992 and June 1998, 70 patients underwent surgery for type A aortic dissection. Fifty-eight patients were discharged from the hospital (17% operative mortality) and were followed by serial MRI for 12 to 90 months after surgery. In all, 436 postoperative MRI examinations were analyzed. In 13 patients (22.5%) no residual intimal flap was identified, whereas 45 patients (77.5%) presented with distal dissection, with a partial thrombosis of the false lumen in 24. The yearly aortic growth rate was maximum in the descending aortic segment (0.37 +/- 0.43 cm) and was significantly higher in the absence of thrombus in the false lumen (0.56 +/- 0.57 cm) (p <0.05). There were 4 sudden deaths, with documented aortic rupture in 2. Sixteen patients underwent reoperation for expanding aortic diameter. In all but 1 patient, a residual dissection was present (in 13 without any thrombosis of the false lumen). Close MRI follow-up in patients after dissection surgical repair can identify the progression of aortic pathology, providing effective prevention of aortic rupture and timely reoperation. Thrombosis of the false lumen appears to be a protective factor against aortic dilation. PMID- 11024404 TI - Effect of cisapride on QT interval in patients with end-stage renal disease. PMID- 11024405 TI - Implantable cardioverter defibrillator therapy for patients with life-threatening ventricular arrhythmias and severe heart failure. PMID- 11024406 TI - Onset heart rate and microvolt t-wave alternans during atrial pacing. PMID- 11024407 TI - Comparison of QT intervals in African-Americans versus Caucasians. PMID- 11024408 TI - Effects of systemic hypertension on arterial dynamics and left ventricular compliance in patients >/=70 years of age. PMID- 11024409 TI - Meta-analysis of effect of beta-blocker therapy on mortality in patients with New York Heart Association class IV chronic congestive heart failure. PMID- 11024410 TI - Use of the thrombolysis in myocardial infarction frame count for the quantitative assessment of transplant-associated arteriosclerosis. PMID- 11024411 TI - Effects of estrogen and the selective estrogen receptor modulator raloxifene on markers of inflammation in postmenopausal women. PMID- 11024412 TI - Submacular surgery trials randomized pilot trial of laser photocoagulation versus surgery for recurrent choroidal neovascularization secondary to age-related macular degeneration: I. Ophthalmic outcomes submacular surgery trials pilot study report number 1. AB - PURPOSE: To report complications and changes in vision during 2 years of follow up of patients with age-related macular degeneration assigned randomly to surgical removal or to laser photocoagulation of subfoveal recurrent neovascular lesions in a pilot trial designed to test methods, to refine estimates of outcome rates, and to project patient accrual rates for a larger multicenter randomized trial to evaluate submacular surgery. PATIENTS AND METHODS: Eligible patients with previous laser photocoagulation of extrafoveal or juxtafoveal choroidal neovascularization secondary to age-related macular degeneration were enrolled at 15 collaborating clinical centers. Assignments to treatment arm were made by personnel at a central coordinating center. Adherence to eligibility criteria and treatment assignment was assessed centrally at a photograph reading center. Patients were examined at 3, 6, 12, and 24 months after treatment for data collection purposes. Outcome measures reported include treatment complications, adverse events, requirements for additional treatment, and 2-year changes in visual acuity from baseline. RESULTS: Of 70 patients enrolled, 36 were assigned to laser photocoagulation and 34 to submacular surgery; all were treated as assigned. One patient in each group died before the 2-year examination. Visual acuity was measured at the 2-year examination for 31 of the surviving patients (89%) in the laser arm and for 28 of the surviving patients (85%) in the surgery arm. The 2-year measurements for 36 of the 59 patients (61%) were made by an examiner masked to treatment assignment and to the identity of the study eye. Improvements and losses of visual acuity were observed in both treatment arms; 20 of 31 study eyes (65%) in the laser arm and 14 of 28 study eyes (50%) in the surgery arm had visual acuity 2 years after enrollment that was better than or no more than 1 line worse than the baseline level. Changes in visual acuity and the size of the central macular lesions from baseline to the 2-year examination were similar in the treatment arms. Few serious complications were observed in either arm at the time of initial treatment; serious adverse events were rare. During follow-up, 11 laser-treated eyes and 18 surgically treated eyes had additional intraocular procedures. CONCLUSIONS: The data from this pilot trial suggest no reason to prefer submacular surgery over laser photocoagulation for treatment of patients with age-related macular degeneration who have lesions similar to those studied in this pilot trial. Any clinical trial designed to compare submacular surgery with laser photocoagulation in eyes with age-related macular degeneration and subfoveal recurrent neovascular lesions must enroll several hundred patients in order to reach a statistically valid conclusion regarding differences between these two methods of treatment with respect to either changes in visual acuity or complication rates. PMID- 11024413 TI - Submacular surgery trials randomized pilot trial of laser photocoagulation versus surgery for recurrent choroidal neovascularization secondary to age-related macular degeneration: II. Quality of life outcomes submacular surgery trials pilot study report number 2. AB - PURPOSE: To summarize findings from health-related quality-of-life interviews with patients who had recurrent subfoveal choroidal neovascularization secondary to age-related macular degeneration and participated in a randomized pilot trial (focal confluent laser photocoagulation versus submacular surgery) that may assist in planning future assessments of health-related quality of life in patients with age-related macular degeneration. PATIENTS AND METHODS: Of 70 patients enrolled in the pilot trial, 54 were interviewed before random assignment to treatment and provided the data analyzed for this report. Patients were scheduled for follow-up interviews at 6, 12, and 24 months after enrollment. The 36-Item Short-Form Health Survey was used in all cases and was administered by an interviewer. The summary scales, Physical Component Summary and Mental Component Summary, were the focus of this analysis. RESULTS: Median Physical Component Summary and Mental Component Summary score at baseline were 47 and 53, respectively. Distributions of scores at baseline and throughout the 2-year follow-up period were similar in the two treatment arms and consistent with those of a sample of the general US population of similar age. Distributions of both Physical Component Summary scores and Mental Component Summary scores for individual patient subgroups at baseline suggested sensitivity to some measures of baseline vision. During 2 years of follow-up interviews, median changes in Physical Component Summary scores were decreases of six points in the laser arm and three points in the surgery arm; in both treatment arms, median change in Mental Component Summary scores was an increase of 2 points. Two-year changes in visual acuity had no discernible effect on Physical Component Summary scores or Mental Component Summary scores; baseline scores were the strongest predictors of 2-year changes in Physical Component Summary and Mental Component Summary scores. CONCLUSIONS: Overall, summary scores from the 36-Item Short-Form Health Survey and changes in scores over 2 years in this pilot trial were consistent with the ophthalmic outcome measures (reported elsewhere) that showed no important differences between the treatment arms. These health-related quality-of-life findings provided assurance that the similarity of ophthalmic outcomes in the two treatment arms did not mask changes potentially important to patients that were not captured as part of the clinical outcome data. PMID- 11024414 TI - Limited inferior macular translocation for the treatment of subfoveal choroidal neovascularization secondary to age-related macular degeneration. AB - PURPOSE: To review a series of patients with age-related macular degeneration undergoing limited macular translocation for the treatment of subfoveal choroidal neovascularization, to determine short-term visual acuity outcomes, to measure amounts of attainable retinal movement, and to identify prognostic factors. METHODS: A retrospective review was conducted on a consecutive series of patients undergoing inferior limited macular translocation with scleral imbrication for the treatment of subfoveal choroidal neovascularization secondary to age-related macular degeneration. The main outcome measures investigated were distance of macular translocation, visual acuity at 3 and 6 months after surgery, change in visual acuity from baseline, and the development of intraoperative and postoperative complications. Univariate and multivariate analyses of a number of potential prognostic factors were undertaken. RESULTS: Macular translocation was achieved in all 102 eyes (101 patients) included in this study. The range of movement varied from 200 to 2,800 microm with a median movement of 1, 200 microm. Nearly 33% of the study group achieved a visual acuity better than 20/100 at 3 months, and 49% achieved this vision at 6 months. At 3 and 6 months, 37% and 48% of the study group, respectively, experienced 2 or more lines of improvement on visual acuity testing, and by 6 months 16% experienced greater than 6 lines of visual improvement. Good baseline vision, achieving the desired amount of macular translocation, a greater amount of macular translocation, and recurrent choroidal neovascularization at baseline were associated with better visual acuities at 3 and 6 months. Poor preoperative vision and the development of complications were associated with worse vision at 3 and 6 months. CONCLUSIONS: Limited macular translocation is a technically feasible procedure that can lead to significant visual improvement and good visual acuity in some patients presenting with subfoveal choroidal neovascularization associated with age-related macular degeneration. A randomized prospective clinical trial of this surgical technique is warranted. PMID- 11024415 TI - The Advanced Glaucoma Intervention Study (AGIS): 7. The relationship between control of intraocular pressure and visual field deterioration.The AGIS Investigators. AB - PURPOSE: To investigate the association between control of intraocular pressure after surgical intervention for glaucoma and visual field deterioration. METHODS: In the Advanced Glaucoma Intervention Study, eyes were randomly assigned to one of two sequences of glaucoma surgery, one beginning with argon laser trabeculoplasty and the other trabeculectomy. In the present article we examine the relationship between intraocular pressure and progression of visual field damage over 6 or more years of follow-up. In the first analysis, designated Predictive Analysis, we categorize 738 eyes into three groups based on intraocular pressure determinations over the first three 6-month follow-up visits. In the second analysis, designated Associative Analysis, we categorize 586 eyes into four groups based on the percent of 6-month visits over the first 6 follow-up years in which eyes presented with intraocular pressure less than 18 mm Hg. The outcome measure in both analyses is change from baseline in follow-up visual field defect score (range, 0 to 20 units). RESULTS: In the Predictive Analysis, eyes with early average intraocular pressure greater than 17.5 mm Hg had an estimated worsening during subsequent follow-up that was 1 unit of visual field defect score greater than eyes with average intraocular pressure less than 14 mm Hg (P =.002). This amount of worsening was greater at 7 years (1.89 units; P <.001) than at 2 years (0.64 units; P =.071). In the Associative Analysis, eyes with 100% of visits with intraocular pressure less than 18 mm Hg over 6 years had mean changes from baseline in visual field defect score close to zero during follow-up, whereas eyes with less than 50% of visits with intraocular pressure less than 18 mm Hg had an estimated worsening over follow-up of 0.63 units of visual field defect score (P =.083). This amount of worsening was greater at 7 years (1.93 units; P <.001) than at 2 years (0.25 units; P =.572). CONCLUSIONS: In both analyses low intraocular pressure is associated with reduced progression of visual field defect, supporting evidence from earlier studies of a protective role for low intraocular pressure in visual field deterioration. PMID- 11024416 TI - Contralateral effect of topical beta-adrenergic antagonists in initial one-eyed trials in the ocular hypertension treatment study. AB - PURPOSE: To evaluate the magnitude of the contralateral effect of topically administered beta-blockers on intraocular pressure. METHODS: The Ocular Hypertension Treatment Study enrolled 1,636 subjects. Of these, 817 subjects were randomized to receive topical ocular hypotensive medication and 819 subjects were randomized to close observation (i.e., no topical medication). We compared the intraocular pressure of the contralateral eye of subjects at the baseline visit and after an initial one-eyed therapeutic trial of topical beta-blockers. We examined differences between baseline and follow-up intraocular pressure in untreated eyes of subjects randomized to close observation. RESULTS: The mean reduction in intraocular pressure in the beta-blocker-treated eyes was -5.9 +/- 3. 4 mm Hg (-22% +/- 12%; Student t test, P <.0001). In the contralateral eyes, mean intraocular pressure reduction was -1.5 +/- 3.0 mm Hg (-5.8% +/- 12%; P <.0001). Of the contralateral eyes, 35% showed a reduction of 3 mm Hg or more, and 10% showed a reduction of 6 mm Hg or more. The contralateral effect of the relatively selective beta-blocker betaxolol did not differ from that of any of the nonselective beta-blockers. Factors associated with the magnitude of the contralateral effect were the degree of intraocular pressure reduction in the treated eye and baseline intraocular pressure of the contralateral eye. In the close observation group, no significant reduction in intraocular pressure was noted between the baseline and follow-up visit. CONCLUSIONS: The contralateral effect is important in clinical practice and in clinical trials when the hypotensive effect of a topical beta-blocker is evaluated by means of a one-eyed therapeutic trial. PMID- 11024417 TI - Retrobulbar hemodynamics in normal-tension glaucoma with asymmetric visual field change and asymmetric ocular perfusion pressure. AB - PURPOSE: To investigate retrobulbar hemodynamics by means of color Doppler imaging in normal-tension glaucoma with asymmetric visual field change and asymmetric ocular perfusion pressure. METHODS: Forty-nine consecutive patients who met the enrollment criteria were enrolled in a prospective study. We measured intraocular pressure at 2-hour intervals for 24 hours and performed color Doppler imaging of the orbital arteries, 48-hour ambulatory blood pressure monitoring, and visual field testing. Color Doppler parameters were compared between subjects with higher ocular perfusion pressure in the eye that had a better mean deviation (concordant group) and worse mean deviation (discordant group). RESULTS: In the eyes with the better mean deviation, the end-diastolic velocity of the ophthalmic artery was significantly slower in the discordant group (P=.0145), while in the eyes with the worse mean deviation, the peak systolic and end-diastolic velocities of the ophthalmic artery were significantly slower and the resistance index of the ophthalmic artery was significantly higher in the discordant group (P=.0395, P=.0088, and P=.0324, respectively). In the concordant group, the end diastolic velocity of the central retinal artery was slower in eyes with a worse mean deviation. CONCLUSION: Retrobulbar hemodynamics are significantly altered in patients with normal-tension glaucoma who have better ocular perfusion pressure in the eye that has a worse mean deviation. PMID- 11024418 TI - Lisch corneal dystrophy is genetically distinct from Meesmann corneal dystrophy and maps to xp22.3. AB - PURPOSE: There is an ongoing discussion whether Lisch corneal dystrophy (band shaped and whorled microcystic dystrophy of the corneal epithelium) represents a disorder that is different from Meesmann corneal dystrophy. The purpose of this study was to evaluate at the molecular level if Lisch and Meesmann corneal dystrophies are genetically distinct. METHODS: We examined at the slit lamp a total of 48 members of a family with an aggregation of Lisch corneal dystrophy. Genomic DNA was extracted from leukocytes of the peripheral blood of seven affected and six unaffected members of this family. Mutational hotspots in the cornea-specific keratin genes K3 and K12 were scanned for mutations by single strand conformation analysis. To test for linkage to the keratin K3 or K12 loci or for X-chromosomal inheritance, six (K3) and four (K12) microsatellite markers each flanking the keratin loci as well as 22 microsatellite markers covering the X-chromosome were typed. Linkage was analyzed using the MLINK and FASTMAP procedures. RESULTS: A total of 19 trait carriers were identified in six generations of the family. No hereditary transmission from father to son was observed. Linkage was excluded for the keratin K3 and K12 genes. Furthermore, single-strand conformation analysis detected no mutations in these genes. Multipoint linkage analysis revealed linkage with a maximum likelihood of the odds (LOD) score of 2.93 at Xp22.3. Linkage was excluded for Xp22.2 to Xqter. CONCLUSIONS: Lisch corneal dystrophy is genetically different from Meesmann corneal dystrophy. Evidence was found for linkage of the gene for Lisch corneal dystrophy to Xp22.3. PMID- 11024419 TI - Episcleritis and scleritis: clinical features and treatment results. AB - PURPOSE: To evaluate the clinical experience with episcleritis and scleritis at a tertiary care eye center. METHODS: Retrospective chart review. RESULTS: One hundred thirty-four patients with scleral inflammation were seen over a 12-year period. Thirty-seven patients had episcleritis, and 97 patients had scleritis. Ocular complications occurred in only 13.5% of patients with episcleritis but in 58.8% of patients with scleritis (P <.0001). No patient with episcleritis had a decrease in visual acuity, whereas 15.9% of patients with scleritis did. Only 16.7% of patients with episcleritis required more than topical corticosteroids for treatment, and these patients required oral nonsteroidal anti-inflammatory drugs. Conversely, 30.4% of patients with scleritis required nonsteroidal anti inflammatory drugs, 31.9% oral prednisone, and 26.1% systemic immunosuppressive drugs (P <.0001). Necrotizing scleritis and posterior scleritis more often were associated with ocular complications, occurring in 91.7% and 85.7%, respectively, than were diffuse anterior scleritis and nodular anterior scleritis (P =.020). Patients with necrotizing scleritis and posterior scleritis were more likely to be treated with oral corticosteroids or immunosuppressive drugs (90% and 100%, respectively) than were patients with diffuse anterior scleritis and nodular anterior scleritis (56.4% and 21.4%, respectively, P =.002). CONCLUSIONS: Scleritis is a severe ocular inflammation, often associated with ocular complications, and nearly always treated with systemic medications. Nearly 60% of these patients will need oral corticosteroids or immunosuppressive drugs to control the disease. PMID- 11024420 TI - Influence of astigmatism on multifocal and monofocal intraocular lenses. AB - PURPOSE: To examine the influence of astigmatism on the visual acuity of patients with multifocal and monofocal intraocular lenses. METHODS: Thirty eyes of 30 patients who underwent five-zone refractive multifocal intraocular lens implantation and 30 eyes of 30 age-matched patients who had monofocal intraocular lens implantation were included. The visual acuities of these patients at 5.0, 3.0, 2.0, 1.0, 0.7, 0.5, and 0.3 m were measured by means of an all-distance vision tester after addition of a cylindrical lens of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 diopters. RESULTS: Mean visual acuity at all distances decreased in proportion to the diopters of astigmatism in both the multifocal and monofocal intraocular lens groups. When astigmatism was 0.5, 1.0, or 1.5 diopters, distance visual acuity in the multifocal group was significantly worse than that in the monofocal group; with astigmatism of 2.0 diopters or more, intermediate visual acuity was also worse in the multifocal group. In contrast, near visual acuity in the multifocal group was significantly better than that in the monofocal group at all astigmatic diopters. When astigmatism was within 1.0 diopter, visual acuity in the multifocal group reached 20/29 at distance and 20/50 at near. Mean contrast sensitivity was also worse in the multifocal group than in the monofocal group. CONCLUSIONS: Both distance and intermediate visual acuity deterioration caused by astigmatism was greater with a multifocal intraocular lens than with a monofocal intraocular lens, whereas near visual acuity was better with the multifocal intraocular lens. When astigmatism was within 1.0 diopter, eyes with a multifocal intraocular lens achieved good visual acuity at both distance and near. PMID- 11024421 TI - Ultrasound biomicroscopy of the anterior segment after congenital cataract surgery. AB - PURPOSE: To evaluate the structure of the anterior segment after congenital cataract surgery with ultrasound biomicroscopy. METHODS: Ultrasound biomicroscopy was performed on 28 eyes of 15 patients after congenital cataract surgery (age range, 8-27 years; mean, 13.1 +/- 1.1 years), of which 26 eyes had pars plana lensectomy and two eyes had phacoemulsification and aspiration, and on 21 eyes of 11 phakic control subjects who were healthy (age range, 6-17 years; mean, 11.5 +/ 1.1 years). The angle-opening distance at 500 microm was measured on a line perpendicular to the trabecular meshwork 500 microm from the scleral spur, and the trabecular-iris angle was measured. RESULTS: Angle-opening distance at 500 microm in the congenital cataract group ranged from 174 to 920 microm (mean, 424 +/- 28 microm), and in the control group it ranged from 260 to 1,119 microm (mean, 643 +/- 57 microm), a difference that was significantly different (P =.0004, Student t test). The trabecular-iris angle in the congenital cataract group ranged from 30.4 to 66.9 degrees (mean, 41.9 +/- 1.6 degrees) and in the control group ranged from 32.1 to 69.8 degrees (mean, 44.9 +/- 2.5 degrees), a difference that was not significantly different (P =.28, Student t test). This means that there is a tendency for high insertion of the iris in the congenital cataract group, and intraocular pressure was elevated after surgery (including glaucoma and ocular hypertension) in 10 (47.6%) of 21 eyes with high insertion. The pars plicata after congenital cataract surgery was significantly flatter than that of control subjects (P =.0057, Student t test). CONCLUSIONS: Ultrasound biomicroscopy is a useful method for evaluating the anterior segment of eyes with congenital cataract, and complications after cataract surgery can be caused by disorganization of the anterior segment. Patients with abnormalities of the anterior segment should be followed up closely for the development of glaucoma. PMID- 11024422 TI - Intraocular pressure and visual field progression in open-angle glaucoma. PMID- 11024424 TI - Transcorneal oxygen therapy for glaucoma associated with sickle cell hyphema. AB - PURPOSE: To study three patients with glaucoma caused by sickle cell hyphema who were successfully treated with transcorneal oxygen therapy. METHODS: Case reports. Three patients with increased intraocular pressure caused by sickle cell hyphema were administered transcorneal oxygen therapy using humidified oxygen at a flow rate that ranged from 1 to 3 l/minute. RESULTS: All three patients had a dramatic reduction in their intraocular pressure within hours of receiving oxygen therapy. No complications were associated with the oxygen therapy. CONCLUSION: Transcorneal oxygen therapy can reduce intraocular pressure in patients with glaucoma from sickle cell hyphema. Further study is warranted to evaluate this new therapy. PMID- 11024423 TI - Guidelines for the use of immunosuppressive drugs in patients with ocular inflammatory disorders: recommendations of an expert panel. AB - PURPOSE: To provide recommendations for the use of immunosuppressive drugs in the treatment of patients with ocular inflammatory disorders. PARTICIPANTS: A 12 person panel of physicians with expertise in ophthalmologic, pediatric, and rheumatologic disease, in research, and in the use of immunosuppressive drugs in patient care. EVIDENCE: Published clinical study results. Recommendations were rated according to the quality and strength of available evidence. PROCESS: The panel was convened in September of 1999 and met regularly through May 2000. Subgroups of the panel summarized and presented available information on specific topics to the full panel; recommendations and ratings were determined by group consensus. CONCLUSIONS: Although corticosteroids represent one of the mainstays in the management of patients with ocular inflammation, in many patients, the severity of the disease, the presence of corticosteroid side effects, or the requirement for doses of systemic corticosteroids highly likely to result in corticosteroid complications supports the rationale for immunosuppressive drugs (for example, antimetabolites, T-cell inhibitors, and alkylating agents) being used in the management of these patients. Because of the potential for side effects, treatment must be individualized and regular monitoring performed. With careful use of immunosuppressive drugs for treatment of ocular inflammatory disorders, many patients will benefit from them either with better control of the ocular inflammation or with a decrease in corticosteroid side effects. PMID- 11024425 TI - Association of autosomal dominantly inherited corneal dystrophies with BIGH3 gene mutations in Japan. AB - PURPOSE: To evaluate the incidence of BIGH3 gene mutations in 164 unrelated Japanese patients with corneal stromal dystrophies with an autosomal dominant trait. METHODS: Data were collected at two major institutions in the eastern and western parts of Japan, where molecular genetic analysis was performed for diagnostic purpose. RESULTS: The incidence of mutations was ranked as follows: 118 patients (72%), the R124H mutation associated with Avellino corneal dystrophy; 23 patients (14%), the R124C mutation associated with lattice corneal dystrophy type 1; and 10 patients (6%), the P501T mutation associated with lattice corneal dystrophy type 3A. CONCLUSION: Avellino corneal dystrophy associated with the R124H mutation is the most common form of corneal stromal dystrophy in Japan. This dystrophy, which is diagnosed histopathologically, has also been called granular corneal dystrophy in Japan. The classification of these diseases according to genetic pathogenesis may be more appropriate than is the use of clinical or histological findings. PMID- 11024426 TI - Penetrating keratoplasty from a temporal approach. AB - PURPOSE: To describe the technique of penetrating keratoplasty using a temporal approach. METHODS: Report of two cases of phakic penetrating keratoplasty performed with the surgeon positioned at the temporal side of the head. RESULTS: The temporal approach improved the surgical exposure, which eliminated the need for a bridal suture, facilitated the placement of the first two cardinal sutures, and facilitated subsequent placement of nasal sutures. CONCLUSION: The temporal approach to penetrating keratoplasty may facilitate suture placement, decrease surgical time, and reduce suture-induced astigmatism. PMID- 11024427 TI - Chronic exposure keratopathy complicating surgical correction of ptosis in patients with chronic progressive external ophthalmoplegia. AB - PURPOSE: To report chronic exposure keratopathy related to surgical ptosis correction in patients with chronic, progressive, external ophthalmoplegia. METHODS: Case reports of three patients with chronic exposure keratopathy following blepharoptosis surgery. RESULTS: We report three patients with chronic progressive external ophthalmoplegia with chronic corneal complications after surgical ptosis repair. All three gave a history of blepharoptosis and extraocular muscle dysfunction. Each presented with chronic corneal ulceration. All had histories suggestive of ophthalmoplegia. Treatment of corneal ulceration necessitated hospitalization and surgical intervention. CONCLUSION: Patients with chronic, progressive, external ophthalmoplegia have little ability to properly protect the eye from exposure and are at risk for corneal damage. A thorough ophthalmic history and examination before ptosis surgery may prevent the corneal complications resulting from surgical intervention. PMID- 11024428 TI - A complication of scleral expansion surgery for treatment of presbyopia. AB - PURPOSE: To report a patient who developed complications from an experimental technique using scleral expansion to treat presbyopia. METHODS: Case report of a 46-year-old woman who underwent scleral expansion surgery on her right eye. Postoperatively, the patient developed chronic pain and swelling that necessitated removal of the scleral expanders. RESULTS: After removal of the scleral expanders, the patient demonstrated a -1.4 diopter myopic shift in the right eye relative to her preoperative refraction. Axial length of the right eye was 1.15 mm longer than of the left, whereas preoperative axial lengths were equal. CONCLUSION: This patient developed scleral thinning with resultant axial lengthening and myopic shift, akin to the phenomenon observed with scleral buckles used to treat retinal detachments. We believe this was responsible for improved near vision in her right eye, rather than any increased accommodative potential purported to result from this operation. PMID- 11024429 TI - Systemic amyloidosis presenting as a mass of the conjunctival semilunar fold. AB - PURPOSE: To report a case of systemic amyloidosis that presented as a mass of the plica semilunaris (semilunar fold) of the conjunctiva. METHODS: Case report. RESULTS: A healthy 56-year-old man developed a fleshy, hemorrhagic lesion of the semilunar fold of the left conjunctiva. Excisional biopsy revealed an acellular lesion of the conjunctival stroma that stained positively for Congo red and showed apple-green birefringence and dichroism with polarization microscopy. The diagnosis was amyloidosis of the semilunar fold of the conjunctiva. Subsequent evaluation disclosed systemic amyloidosis. The patient remains asymptomatic. CONCLUSIONS: Although conjunctival amyloidosis usually occurs as a localized lesion without systemic involvement, it can be the initial sign of systemic amyloidosis. PMID- 11024430 TI - Comparison of visual function after foveal translocation with 360 degrees retinotomy and with scleral shortening in a patient with bilateral myopic neovascular maculopathy. AB - PURPOSE: To compare the visual outcome after foveal translocation by scleral shortening and that after 360 degrees retinotomy with extraocular muscle surgery in a patient with bilateral myopic neovascular maculopathy. METHODS: Case report. RESULTS: A 52-year-old woman with bilateral myopic neovascular maculopathy underwent foveal translocation with scleral shortening in the left eye, and visual acuity improved from 20/70 to 20/30. However, choroidal neovascularization recurred, and the final visual acuity was 20/40 after excision of the choroidal neovascularization. Foveal translocation with 360 degrees retinotomy was performed on the right eye, and visual acuity improved from 20/150 to 20/30. The critical print size was better, and the retinal sensitive area was larger in the right eye. CONCLUSION: The better reading ability shown by foveal translocation by a 360 degrees retinotomy compared with scleral shortening may stem from a larger retinal sensitive area obtained by this method. PMID- 11024431 TI - Decompensation of a congenital retinal macrovessel with arteriovenous communications induced by repetitive rollercoaster rides. AB - PURPOSE: To describe a congenital retinal venous macrovessel that communicates with a cilioretinal artery and a retinal artery, and to report how this vascular anomaly decompensated as a result of repetitive rollercoaster rides. METHODS: Case report with serial fundus photography and fluorescein angiography. RESULTS: After a short period of intensive rollercoaster rides, a 19-year-old woman complained of reduced vision in one eye. Funduscopy and fluorescein angiography revealed a venous congenital retinal macrovessel with arteriovenous communications, and retinal exudation was visible at the termination of the anomalous vessel. Exudation resolved, and acuity recovered after a period of avoidance of rollercoaster rides. CONCLUSION: This case represents the first report of a retinal artery and a cilioretinal artery communicating with a congenital retinal macrovessel, and it suggests that such patients are at increased risk of retinal vascular decompensation if involved in activities associated with changes in g-forces, such as bungee jumping or rollercoaster rides. PMID- 11024432 TI - Neovascular glaucoma as a complication of retinal vasculitis in Crohn disease. AB - PURPOSE: To report a case of neovascular glaucoma as a complication of retinal vasculitis in Crohn disease. METHODS: Case report with fluorescein angiogram. RESULTS: A 62-year-old man with biopsy-proven Crohn disease presented with bilateral uveitis, bilateral iris new vessels, and neovascular glaucoma in the left eye. Fluorescein angiography revealed signs of retinal vasculitis and capillary nonperfusion in both eyes. CONCLUSION: Crohn disease may be associated with retinal vasculitis and, thus, neovascular glaucoma. A satisfactory result can be achieved by using corticosteroids to control the retinal vascular inflammation, by applying panretinal photocoagulation and by controlling the increased intraocular pressure surgically. PMID- 11024434 TI - Ocular myasthenia and nitrofurantoin. AB - PURPOSE: To report a case of ocular myasthenia associated with administration of nitrofurantoin. METHODS: Case report and review of the literature. RESULTS: While taking nitrofurantoin after urinary tract surgery, a 10-year-old female developed diplopia and ptosis. Sleep test confirmed ocular myasthenia. Signs and symptoms resolved after discontinuation of the drug. CONCLUSION: Ocular myasthenia may be associated with nitrofurantoin administration. PMID- 11024433 TI - Orbital compression syndrome after orbital extravasation of X-ray contrast material. AB - PURPOSE: To report the orbital compression syndrome after orbital extravasation of x-ray contrast material during catheterization of the left middle meningeal artery. METHODS: Case report. RESULTS: A 61-year-old woman had profound loss of vision, pain, and proptosis of her left eye immediately after catheterization of the left middle meningeal artery. Computed tomography (CT) revealed that contrast material had extravasated into the orbit. Her symptoms improved with lateral canthotomy and cantholysis and resolved totally within 24 hours without the need for an orbital surgical procedure to remove the contrast material. CONCLUSION: This report describes an unusual cause of the orbital compression syndrome. PMID- 11024435 TI - Ocular motor signs in an infant with carbohydrate-deficient glycoprotein syndrome type Ia. AB - PURPOSE: To document the evolution of ocular motor abnormalities in an infant with carbohydrate-deficient glycoprotein syndrome. METHODS: Case report. An infant with carbohydrate-deficient glycoprotein syndrome type 1a underwent magnetic resonance imaging and infrared eye movement recording. RESULTS: A 10 month-old male with carbohydrate-deficient glycoprotein syndrome type Ia had rapid horizontal oscillations of the eyes when startled or awakened from sleep. Clinical examination confirmed this finding and disclosed congenital ocular motor apraxia with a reduced vestibulo-ocular reflex. Infrared eye movement recording showed ocular flutter and square wave jerks superimposed on a horizontal pendular nystagmus. Magnetic resonance imaging showed diffuse cerebellar hypoplasia. CONCLUSION: Carbohydrate-deficient glycoprotein syndrome type Ia can be associated with multiple cerebellar eye signs including ocular flutter, square wave jerks, and congenital ocular motor apraxia. PMID- 11024437 TI - Practice parameter: the role of corticosteroids in the management of acute monosymptomatic optic neuritis. report Of the quality standards subcommittee of the american academy of neurology(1) PMID- 11024436 TI - Pachymeningitis with pseudo-Foster Kennedy syndrome. AB - PURPOSE: To report a case of pachymeningitis with pseudo-Foster Kennedy syndrome in a patient who was positive for perinuclear antineutrophil cytoplasmic antibody. METHODS: Case report. A 44-year-old man was examined for headache and diplopia. RESULTS: Left eye showed limitation of abduction. Ocular fundus, computed tomography (CT), and magnetic resonance imaging (MRI) of the head and orbits were normal. The diplopia subsided spontaneously. Six months later, he noticed sudden visual loss in the left eye. The left eye showed optic disk atrophy and episcleritis, and the right eye showed papilloedema. Computed tomography and MRI exhibited thickened dura mater. Serum perinuclear antineutrophil cytoplasmic antibody level was highly increased. CONCLUSION: Pachymeningitis with ocular involvement sometimes requires repeated CT or MRI for diagnosis. This disorder may be caused by microvasculitis. PMID- 11024438 TI - Proton beam irradiation of subfoveal choroidal neovascularisation in age-related macular degeneration(1) PMID- 11024440 TI - Monovision contact lens use in the aviation environment: a report of a contact lens-related aircraft accident(1) PMID- 11024439 TI - Unnatural injuries(1) PMID- 11024441 TI - Tactile spatial resolution in blind braille readers(1) PMID- 11024442 TI - Radiotherapy for subfoveal neovascularization associated with pathological myopia: a pilot study(1) PMID- 11024443 TI - Reconstruction of damaged corneas by transplantation of autologous limbal epithelial cells(1) PMID- 11024444 TI - The new zealand cataract and refractive surgery survey 1997/1998(1) PMID- 11024446 TI - A hypothetical scheme for the brainstem control of vertical gaze(1) PMID- 11024445 TI - Successful transplantation of bioengineered tissue replacements in patients with ocular surface disease(1) PMID- 11024447 TI - Lysostaphin treatment of methicillin-resistant staphylococcus aureus keratitis in the rabbit(1) PMID- 11024448 TI - The effect of contrast on reading speed in dyslexia(1) PMID- 11024449 TI - Emerging perspectives on glaucoma: highlights of a roundtable discussion. AB - PURPOSE: To present emerging perspectives on open-angle glaucoma related to patient compliance, current drug therapy, and treatment options. METHODS: Roundtable discussion. RESULTS: The panel discussion identified the following issues about patient compliance:1. It is a common problem in clinical practice and is often underappreciated by physicians.2. It is difficult for clinicians to detect.3. There are several risk factors that increase the probability of noncompliance. 4. Compliance can be improved through education, communication, improvement of the doctor-patient relationship, compliance aids, and simplifying the treatment regimen. CONCLUSION: Patients' compliance with the therapeutic regimen for open-angle glaucoma is very important to the eventual outcome of their disease treatment. Physicians can improve their patients' compliance by a few simple, cost-effective interventions in the clinical setting. When considering different agents as first-line therapies, once-a-day dosing would be optimal for enhancing compliance. PMID- 11024450 TI - Target cells for methylsulphonyl-2,6-dichlorobenzene in the olfactory mucosa in mice. AB - Previously we reported that methylsulphonyl-2,6-dichlorobenzene, 2, 6-(diCl MeSO(2)-B), was irreversibly bound to the olfactory mucosa of mice and induced necrosis of the Bowman's glands with subsequent neuroepithelial degeneration and detachment. In this study, autoradiography and histopathology were used to determine tissue-localization and toxicity of 2,6-(diCl-MeSO(2)-B) in the olfactory mucosa of control mice and animals pretreated with cytochrome P450 (CYP) and glutathione (GSH) modulators. The Bowman's glands of the olfactory mucosa were the major target sites of non-extractable binding of 2,6-(diCl-(14)C MeSO(2)-B), whereas the olfactory neuroepithelium and nerve bundles showed only background levels of silver grains. Metyrapone pretreatment slightly decreased binding in the Bowman's glands and markedly decreased toxicity in the olfactory mucosa after 2,6-(diCl-MeSO(2)-B) administration. These results support that a CYP-mediated activation of 2, 6-(diCl-MeSO(2)-B) takes place in the Bowman's glands giving rise to toxic reactive intermediates. In mice pretreated with the GSH-depleting agent phorone, a marked increase of irreversible binding of 2,6 (diCl-(14)C-MeSO(2)-B) in the Bowman's glands was observed. Tape-section autoradiograms also revealed a significant increase of uptake of radioactivity in the olfactory bulb. As determined by histopathology, GSH-depletion increased both the extent and severity of the lesion in the mucosa. These results imply that 2,6 (diCl-MeSO(2)-B)-reactive intermediates are conjugated with GSH. The amount of irreversible binding and toxicity in the olfactory mucosa seems to be associated with the level of 2, 6-(diCl-MeSO(2)-B)-reactive intermediates. PMID- 11024451 TI - Suppressive effect of penta-acetyl geniposide on the development of gamma glutamyl transpeptidase foci-induced by aflatoxin B(1) in rats. AB - The suppressive effects of penta-acetyl geniposide, (Ac)(5)-GP, on the hepatotoxic lesions-induced by aflatoxin B(1) (AFB(1)) were investigated in male Wistar rats. Rats were divided into six groups: groups I and II served as normal and solvent control, respectively; group III was given AFB(1) (2 mg/kg body weight) alone; group IV was given (Ac)(5)-GP (2 mg/kg) alone; and groups V and VI received both AFB(1) (2 mg/kg body weight) and (Ac)(5)-GP (1 mg and 2 mg/kg body weight, respectively). Rats received treatments for 8 weeks, then were maintained on basal diet for 32 weeks. At the end of the experiment (week 40), the liver lesions (e.g. fatty change, eosinophilic and bile duct dilation) and preneoplastic changes in rats of groups V and VI were reduced when they were compared with group III. There were no liver lesions and preneoplastic changes in rats treated with (Ac)(5)-GP alone. Although no differences in the total number of gamma-glutamyl transpeptidase (GGT)-positive foci was observed between the groups treated with AFB(1) along with or without (Ac)(5)-GP, the treatment of (Ac)(5)-GP significantly reduced the number of AFB(1)-induced GGT positive foci (with diameter larger than 0.3 mm). These results indicated that the protective effect of (Ac)(5)-GP on early hepatocarcinogenesis-induced by AFB(1) was associated with the inhibition of GGT foci development. PMID- 11024452 TI - Relationship of MTT reduction to stimulants of muscle metabolism. AB - MTT, a positively charged tetrazolium salt, is widely used as an indicator of cell viability and metabolism and has potential for histochemical identification of tissue regions of hypermetabolism. In the present study, MTT was infused in the constant-flow perfused rat hindlimb to assess the effect of various agents and particularly vasoconstrictors that increase muscle metabolism. Reduction of MTT to the insoluble formazan in muscles assessed at the end of experiments was linear over a 30 min period and production rates were greater in red fibre types than white fibre types. The vasoconstrictors, norepinephrine (100 nM) and angiotensin (10 nM) decreased MTT formazan production in all muscles but increased hindlimb oxygen uptake and lactate efflux. Veratridine, a Na(+) channel opener that increases hindlimb oxygen uptake and lactate efflux without increases in perfusion pressure, also decreased MTT formazan production. Membrane stabilizing doses (100 microM) of (+/-)-propranolol reversed the inhibitory effects of angiotensin and veratridine on MTT formazan production. Muscle contractions elicited by stimulation of the sciatic nerve, reversed the norepinephrine-mediated inhibitory effects on MTT formazan production, even though oxygen consumption and lactate efflux were further stimulated. Stimulation of hindlimb muscle oxygen uptake by pentachlorophenol, a mitochondrial uncoupler, was not associated with alterations in MTT formazan production. It is concluded that apart from muscle contractions MTT formazan production does not increase with increased muscle metabolism. Since the vasoconstrictors angiotensin and norepinephrine as well as veratridine activate Na(+) channels and the Na(+)/K(+) pump, energy required for Na(+) pumping may be required for MTT reduction. It is unlikely that vasoconstrictors that stimulate oxygen uptake do so by uncoupling respiration. PMID- 11024453 TI - DNA adduct formation by Fusarium culture extracts: lack of role of fusarin C. AB - Fusarium fungi have been shown to infect corn and other crops worldwide, and have a significant impact on human health through loss of crops or contamination of food with mycotoxins. Isolates of Fusarium fungi from an area of South Africa with high incidence of esophageal cancer have been shown to induce esophageal and liver cancer in rats. Several isolates of Fusarium fungi were grown on corn to determine if genotoxic products were produced. We report the incubation of methanol extracts of Fusarium verticillioides cultures with DNA in the presence of rat liver fractions (S9) resulted in the formation of a unique DNA adduct that was detected by (32)P-postlabeling. Fusarin C was purified from cultures of Fusarium verticillioides RRC 415, and was not responsible for the formation of the DNA adduct. Treatment of the methanolic extracts with ultraviolet B radiation reduced the fusarin C content in the extract; however, this had no effect on the formation of the DNA adduct following incubation of the extract with DNA and S9. The unique DNA adduct was formed following the incubation of several Fusarium verticillioides isolates from the US and South Africa, while extracts of cultures of Fusarium graminearium and Fusarium sacchari isolates formed very little of the DNA adduct when incubated with DNA and S9. These data suggest that neither fusarin C nor any of its metabolites are responsible for formation of the DNA adduct, and that an unidentified compound is present in F. verticillioides cultures that forms a DNA adduct, and may be important in the etiology of human esophageal cancer. PMID- 11024454 TI - Differential feedback regulation of the MAPK cascade underlies the quantitative differences in EGF and NGF signalling in PC12 cells. AB - Although epidermal growth factor (EGF) induces transient activation of Ras and the mitogen-activated protein kinase (MAPK) cascade in PC12 cells, whereas nerve growth factor (NGF) stimulates sustained activation, the basis for these contrasting responses is not known. We have developed a computer simulation of EGF-induced MAPK cascade activation, which provides quantitative evidence that feedback inhibition of the MAPK cascade is the most important factor in determining the duration of cascade activation. Hence, we propose that the observed quantitative differences in EGF and NGF signalling can be accounted for by differential feedback regulation of the MAPK cascade. PMID- 11024455 TI - Self-complementary motifs (SCM) in alpha-crystallin small heat shock proteins. AB - Small heat shock proteins (sHsp) have been implicated in many cell processes involving the dynamics of protein-protein interactions. Two unusual sequences containing self-complementary motifs (SCM) have been identified within the conserved alpha-crystallin domain of sHsps. When two SCMs are aligned in an anti parallel direction (N to C and C to N), the charged or polar residues form either salt bridges or hydrogen bonds while the non-polar residues participate in hydrophobic interactions. When aligned in reverse order, the residues of these motifs in alpha-crystallin subunits form either hydrophobic and/or polar interactions. Homology based molecular modeling of the C-terminal domain of alpha crystallin subunits using the crystal structure of MjHSP16.5 suggests that SCM1 and 2 participate in stabilizing secondary structure and subunit interactions. Also there is overwhelming evidence that these motifs are important in the chaperone-like activity of alpha-crystallin subunits. These sequences are conserved and appear to be characteristic of the entire sHsp superfamily. Similar motifs are also present in the Hsp70 family and the immunoglobulin superfamily. PMID- 11024456 TI - Interaction of different oligomeric states of hexameric DNA-helicase RepA with single-stranded DNA studied by analytical ultracentrifugation. AB - Analytical ultracentrifugation was used to determine the molecular mass, M, of hexameric DNA-helicase RepA at pH 5.8 and 7.6. At pH 7.6, a molecular mass of 179.5+/-2.6 kDa was found, consistent with the known hexameric state of RepA, (RepA)(6). At pH 5.8, (RepA)(6) associates to form a dimer with a molecular mass of 366.2+/-4.1 kDa. Analytical ultracentrifugation was also applied to characterize the interaction of single-stranded DNA (ssDNA) with the two different oligomeric states of (RepA)(6) at pH 5.8 and 7.6. The dissociation constants, K(d), for the equilibrium binding of (dA)(30) to the (RepA)(6) dimer at pH 5.8 and to (RepA)(6) at pH 7.6 were determined at 10 degrees C in the presence of 0.5 mM ATPgammaS, 10 mM MgCl(2) and 60 mM NaCl as K(d5.8)=0.94+/-0.13 microM at pH 5.8 and K(d7. 6)=25.4+/-6.4 microM at pH 7.6. The stoichiometries, n, for the two complexes (dA)(30)/(RepA)(6) dimer and (dA)(30)/(RepA)(6) at pH 5.8 and 7.6 were calculated from the corresponding binding curves. At pH 5.8 one (dA)(30) molecule was bound per (RepA)(6) dimer, while at pH 7.6 one (dA)(30) molecule was bound to one (RepA)(6). Binding curves were compatible with a single ssDNA binding site present on the (RepA)(6) dimer and on (RepA)(6), respectively, with no indication of cooperativity. (RepA)(6) tends to form larger aggregates under acidic conditions (pH<6.0) which are optimal for ssDNA binding. In contrast, at pH 5.8 in the presence of 60 mM NaCl, only the (RepA)(6) dimer was observed both in the absence and presence of (dA)(30). PMID- 11024457 TI - Expression of different coding sequences in cell-free bacterial and eukaryotic systems indicates translational pausing on Escherichia coli ribosomes. AB - Five different coding sequences of bacterial or eukaryotic origin in plasmids under the T7 promoter were expressed in a cell-free system derived from Escherichia coli. Translation on E. coli ribosomes resulted in a full-length product only in four of the five coding sequences tested. A unique pattern of less than full-length polypeptides was generated in each case. Many of these polypeptides on E. coli ribosomes reacted with a puromycin derivative, cytidylic acid-puromycin, which was radioactively labeled. Thus these incomplete polypeptides can be defined as nascent peptides bound to the ribosomal P site. Certain nascent peptides could be shifted into full-length protein indicating that they resulted from translational pausing. In contrast to these results, expression of the same coding sequences in a wheat germ or reticulocyte cell-free system resulted in a 80-90% full-length product with no evidence for nascent polypeptides and translational pausing. PMID- 11024458 TI - Neuronal nicotinic acetylcholine receptors of Drosophila melanogaster: the alpha subunit dalpha3 and the beta-type subunit ARD co-assemble within the same receptor complex. AB - Dalpha3 is a functional alpha-subunit of Drosophila melanogaster nicotinic acetylcholine receptors (nAChRs). Here, we produced Dalpha3-specific antibodies to study which other nAChR subunits can co-assemble with Dalpha3 in receptor complexes of the Drosophila nervous system. Immunohistochemical studies revealed that Dalpha3 is co-distributed with the beta-subunit ARD in synaptic neuropil regions of the optic lobe. Both subunits can be co-purified by alpha-bungarotoxin affinity chromatography. Dalpha3 antibodies co-immunoprecipitate Dalpha3 and ARD proteins and, vice versa, anti-ARD antibodies co-precipitate ARD and Dalpha3. These data demonstrate that one type of fly nAChRs includes these two subunits as integral components. PMID- 11024459 TI - Mechanisms of resistance of normal cells to TRAIL induced apoptosis vary between different cell types. AB - Resistance of normal cells to tumour necrosis factor related apoptosis inducing ligand (TRAIL) induced apoptosis is believed to be mediated by expression of two decoy receptors. Here we show that the expression and localisation of TRAIL receptors (TRAIL-Rs) vary between different cells and that resistance to TRAIL is mediated by different mechanisms. The decoy receptor, TRAIL-R3, appeared important in protection of endothelial cells, whereas lack of surface death receptor expression and as yet unknown intracellular inhibitor(s) of apoptosis downstream of caspase-3 may play a major role in protection of melanocytes and fibroblasts from TRAIL induced apoptosis, respectively. Differential subcellular location of decoy receptors may be an important determinant of their effectiveness in different types of normal cells. PMID- 11024460 TI - Role of insulin receptor substrate-2 in interleukin-9-dependent proliferation. AB - Interleukin-9 (IL-9) stimulation results in JAK, STAT and IRS1/2 phosphorylation. The role of IRS adaptor proteins in IL-9 signaling is not clear. We show that IL 9 induces IRS2 phosphorylation and association with phosphatidylinositol-3 kinase (PI 3-K) p85 subunit in TS1 cells and BaF/9R cells, which proliferate upon IL-9 stimulation. We observed a PI 3-K-dependent phosphorylation of protein kinase B (PKB) in TS1 cells, but not in BaF/9R, nor in other IL-9-dependent cell lines. Finally, 32D cells that were transfected with the IL-9 receptor but lack IRS expression survived in the presence of IL-9. Ectopic IRS1 expression allowed for IL-9-induced proliferation, in the absence of significant PKB phosphorylation. PMID- 11024461 TI - Serine base-exchange in rat liver nuclei. AB - It has been shown that the incorporation of [(14)C]serine into phosphatidylserine (PS) in isolated rat liver nuclei is intrinsic to this organelle as attested by marker enzyme activity. Serine incorporation into PS was the highest in nuclei depleted of the outer membrane of the nuclear envelope (nucleoplasts) and negligible in the outer membrane. Trypsin treatment of nucleoplasts caused a strong inactivation of PS synthesis and only a moderate one of the NAD pyrophosphorylase activity, the marker enzyme of the inner nuclear membrane. We suggest that the serine base-exchange enzyme is located in the inner membrane of the nuclear envelope and accessible from the periplasmic surface of this membrane. PMID- 11024462 TI - Alternative splicing regulates the nuclear or cytoplasmic localization of dystrophin Dp71. AB - The subcellular distribution of Dp71 isoforms alternatively spliced for exon 71 and/or 78 was examined. The cDNA sequence of each variant was fused to the C terminus of the green fluorescent protein and the constructs were transfected transiently in the cell lines HeLa, C2C12 and N1E-115. The subcellular distribution of the fused proteins was determined by confocal microscope analysis. The Dp71 isoform lacking the amino acids encoded by exons 71 and 78 was found exclusively in the cytoplasm whereas the variants containing the amino acids encoded by exon 71 and/or exon 78 show a predominant nuclear localization. The nuclear localization of Dp71 provides a new clue towards the establishment of its cellular function. PMID- 11024463 TI - Characterisation of new intracellular membranes in Escherichia coli accompanying large scale over-production of the b subunit of F(1)F(o) ATP synthase. AB - Recombinant membrane proteins in Escherichia coli are either expressed at relatively low level in the cytoplasmic membrane or they accumulate as inclusion bodies. Here, we report that the abundant over-production of subunit b of E. coli F(1)F(o) ATP synthase in the mutant host strains E. coli C41(DE3) and C43(DE3) is accompanied by the proliferation of intracellular membranes without formation of inclusion bodies. Maximal levels of proliferation of intracellular membranes were observed in C43(DE3) cells over-producing subunit b. The new proliferated membranes contained all the over-expressed protein and could be recovered by a single centrifugation step. Recombinant subunit b represented up to 80% of the protein content of the membranes. The lipid:protein ratios and phospholipid compositions of the intracellular membranes differ from those of bacterial cytoplasmic membranes, and they are particularly rich in cardiolipin. PMID- 11024464 TI - Membrane interactions and alignment of structures within the HIV-1 Vpu cytoplasmic domain: effect of phosphorylation of serines 52 and 56. AB - The cytoplasmic domain of the HIV-1 accessory protein Vpu is involved in the binding and degradation of the viral receptor CD4. In order to analyze previous structural models in the context of membrane environments, regions of Vpu(CYTO) incorporating particular conformational features have been synthesized and labelled with (15)N at selected backbone amides. Well-oriented proton-decoupled (15)N solid-state NMR spectra with (15)N chemical shifts at the most upfield position indicate that the amphipathic helix within [(15)N-Leu 45]-Vpu(27-57) strongly interacts with mechanically aligned POPC bilayers and adopts an orientation parallel to the membrane surface. No major changes in the topology of this membrane-associated amphipathic helix were observed upon phosphorylation of serine residues 52 and 56, although this modification regulates biological function of Vpu. In contrast, [(15)N-Ala 62]-Vpu(51-81) exhibits a pronounced (15)N chemical shift anisotropy. PMID- 11024465 TI - H(+)/Ca(2+) exchange driven by the plasma membrane Ca(2+)-ATPase of Arabidopsis thaliana reconstituted in proteoliposomes after calmodulin-affinity purification. AB - The plasma membrane Ca(2+)-ATPase was purified from Arabidopsis thaliana cultured cells by calmodulin (CaM)-affinity chromatography and reconstituted in proteoliposomes by the freeze-thaw sonication procedure. The reconstituted enzyme catalyzed CaM-stimulated 45Ca(2+) accumulation and H(+) ejection, monitored by the increase of fluorescence of the pH probe pyranine entrapped in the liposomal lumen during reconstitution. Proton ejection was immediately reversed by the protonophore FCCP, indicating that it is not electrically coupled to Ca(2+) uptake, but it is a primary event linked to Ca(2+) uptake in the form of countertransport. PMID- 11024466 TI - Novel sugar-binding specificity of the type XIII xylan-binding domain of a family F/10 xylanase from Streptomyces olivaceoviridis E-86. AB - The type XIII xylan-binding domain (XBD) of a family F/10 xylanase (FXYN) from Streptomyces olivaceoviridis E-86 was found to be structurally similar to the ricin B chain which recognizes the non-reducing end of galactose and specifically binds to galactose containing sugars. The crystal structure of XBD [Fujimoto, Z. et al. (2000) J. Mol. Biol. 300, 575-585] indicated that the whole structure of XBD is very similar to the ricin B chain and the amino acids which form the galactose-binding sites are highly conserved between the XBD and the ricin B chain. However, our investigation of the binding abilities of wt FXYN and its truncated mutants towards xylan demonstrated that the XBD bound xylose-based polysaccharides. Moreover, it was found that the sugar-binding unit of the XBD was a trimer, which was demonstrated in a releasing assay using sugar ranging in size from xylose to xyloheptaose. These results indicated that the binding specificity of the XBD was different from those of the same family lectins such as the ricin B chain. Somewhat surprisingly, it was found that lactose could release the XBD from insoluble xylan to a level half of that observed for xylobiose, indicating that the XBD also possessed the same galactose recognition site as the ricin B chain. It appears that the sugar-binding pocket of the XBD has evolved from the ancient ricin super family lectins to bind additional sugar targets, resulting in the differences observed in the sugar-binding specificities between the lectin group (containing the ricin B chain) and the enzyme group. PMID- 11024467 TI - Crystal structure of S-glutathiolated carbonic anhydrase III. AB - S-Glutathiolation of carbonic anhydrase III (CAIII) occurs rapidly in hepatocytes under oxidative stress. The crystal structure of the S-glutathiolated CAIII from rat liver reveals covalent adducts on cysteines 183 and 188. Electrostatic charge and steric contacts at each modification site inversely correlate with the relative rates of reactivity of these cysteines toward glutathione (GSH). Diffuse electron density associated with the GSH adducts suggests a lack of preferred bonding interactions between CAIII and the glutathionyl moieties. Hence, the GSH adducts are available for binding by a protein capable of reducing this mixed disulfide. These properties are consistent with the participation of CAIII in the protection/recovery from the damaging effects of oxidative agents. PMID- 11024468 TI - An analysis of the lamellar structure of sea urchin egg cortical granules using X ray scattering. AB - Cortical granules (CGs) are secretory vesicles associated with egg and oocyte plasma membranes that undergo exocytosis at fertilisation. In the sea urchin Strongylocentrotus purpuratus, the internal organisation of these CGs exhibits a lamellar-type morphology. The different lamellar layers correspond to proteoglycans, structural proteins and enzymes required for fertilisation envelope assembly and modification of the post-fertilisation egg surface. We have studied the lamellar structure of CGs using X-ray scattering and reveal the contrast density variation of the lamellae in the native state. The structure of functionally competent CGs in situ differs significantly from that determined by electron microscopic studies. We observed a strong periodicity of the lamellar structure of 280 A as opposed to the 590 A repeat observed previously. Fusion of the CGs produced a loss of the lamellar repeat and the development of a broad peak corresponding to a 20 A periodicity that may be indicative of the molecular packing in the resulting hydrated gel structure. PMID- 11024469 TI - The photoreceptor protein of Euglena gracilis. AB - We isolated the photoactive protein Erh, isolated from the photoreceptor of the unicellular photosynthetic flagellate Euglena gracilis. It is a 27 kDa protein with a photocycle resembling that of sensory rhodopsin, but with at least one stable intermediate. We recorded the absorption spectrum of the parent form of this protein both under native form and in the presence of hydroxylamine and sodium borohydride, and the fluorescence spectra of both the parent and intermediate forms. We suggest that Erh is a rhodopsin-like protein and propose a simple photocycle. This protein shows optical bistability, without thermal deactivation. PMID- 11024470 TI - Resonance Raman spectroscopic study of the tryptic 39-kDa fragment of phytochrome. AB - The 39-kDa fragment of oat phytochrome phyA, obtained by tryptic digestion at the amino acids 65 and 425, was studied by resonance Raman spectroscopy. The parent state P(r) reveals far-reaching similarities with that of the native phytochrome implying that the structures of the tetrapyrrole chromophore and its immediate protein environment are not affected by the proteolysis. However, the resonance Raman spectrum of the final product of the P(r) phototransformation, denoted as P(bl), is more closely related to that of the P(fr) precursor of the native phytochrome, i.e. meta-R(C), rather than to that of P(fr) itself. The resonance Raman spectra indicate a high conformational flexibility of the chromophore in P(bl) so that, unlike in P(fr), the tetrapyrrole rings C and D adopt a largely coplanar conformation. The protein interactions with ring D of the chromophore, which in the native phytochrome stabilize the specific chromophore structure of P(fr), cannot be established in the 39-kDa fragment due to the lack of the major C-terminal part of the protein. These findings, furthermore, support the view that the meta-R(C)-->P(fr) transition is associated with a coupling of chromophore and protein structural changes that represent crucial events for the photoactivation of phytochrome. PMID- 11024471 TI - Laminin-induced PC-12 cell differentiation is inhibited following laser inactivation of cellular prion protein. AB - Prions, the etiological agents for infectious degenerative encephalopathies, act by inducing structural modifications in the cellular prion protein (PrPc). Recently, we demonstrated that PrPc binds laminin (LN) and that this interaction is important for the neuritogenesis of cultured hippocampal neurons. Here we have used the PC-12 cell model to explore the biological role of LN-PrPc interaction. Antibodies against PrPc inhibit cell adhesion to LN-coated culture plaques. Furthermore, chromophore-assisted laser inactivation of cell surface PrPc perturbs LN-induced differentiation and promotes retraction of mature neurites. These results point out to the importance of PrPc as a cell surface ligand for LN. PMID- 11024472 TI - Mutilation of RNA phage Qbeta virus-like particles: from icosahedrons to rods. AB - Icosahedral virus-like particles (VLPs) of RNA phage Qbeta are stabilized by four disulfide bonds of cysteine residues 74 and 80 within the loop between beta strands F and G (FG loop) of the monomeric subunits, which determine the five fold and quasi-six-fold symmetry contacts of the VLPs. In order to reduce the stability of Qbeta VLPs, we mutationally converted the amino acid stretch 76 ANGSCD-81 within the FG loop into the 76-VGGVEL-81 sequence. It led to production in Escherichia coli cells of aberrant rod-like Qbeta VLPs, along with normal icosahedral capsids. The length of the rod-like particles exceeded 4-30 times the diameter of icosahedral Qbeta VLPs. PMID- 11024473 TI - The antennapedia peptide penetratin translocates across lipid bilayers - the first direct observation. AB - The potential use of polypeptides and oligonucleotides for therapeutical purposes has been questioned because of their inherently poor cellular uptake. However, the 16-mer oligopeptide penetratin, derived from the homeodomain of Antennapedia, has been reported to enter cells readily via a non-endocytotic and receptor- and transporter-independent pathway, even when conjugated to large hydrophilic molecules. We here present the first study where penetratin is shown to traverse a pure lipid bilayer. The results support the idea that the uptake mechanism involves only the interaction of the peptide with the membrane lipids. Furthermore, we conclude that the translocation does not involve pore formation. PMID- 11024474 TI - Drosophila AD3 mutation of synaptotagmin impairs calcium-dependent self oligomerization activity. AB - Genetic analysis of a Drosophila synaptotagmin (Syt) I mutant (AD3) has revealed that Tyr-334 within the C2B domain is essential for efficient Ca(2+)-dependent neurotransmitter release. However, little is known as to why a missense mutation (Tyr-334-Asn) disrupts the function of the C2B domain at the molecular level. Here, we present evidence that a Tyr-312 to Asn substitution in mouse Syt II, which corresponds to the Drosophila AD3 mutation, completely impairs Ca(2+) dependent self-oligomerization activity mediated by the C2B domain but allows partial interaction with wild-type proteins in a Ca(2+)-dependent manner. This observation is consistent with the fact that the AD3 allele is homozygous lethal but complements another mutant phenotype. We also showed that the Ca(2+) dependent C2B self-oligomerization is inhibited by inositol 1,3,4, 5 tetrakisphosphate, a potent inhibitor of neurotransmitter release. All of these findings strongly support the idea that self-oligomerization of Syt I or II is essential for neurotransmitter release in vivo. PMID- 11024475 TI - P53 null mice: damaging the hypothesis? AB - P53 is extremely well characterised as a tumour suppressor gene, and many activities have been attributed to it which are consistent with this function. However, despite being the subject of intense study it still remains unclear precisely which of these functions is crucial to its in vivo role as a tumour suppressor gene. This is particularly true of its role in the induction of apoptosis. The original observation of p53-dependent apoptosis gave rise to the following hypothesis: namely, that p53 deficiency leads to a persistence of DNA damaged cells which are the potential founders of malignancy. This review summarises the data for and against this hypothesis, with specific emphasis on data obtained from studies of the murine intestine. What emerges from these studies is a complex picture, where data can be obtained in support of this hypothesis, but there are many circumstances which exist where it is not supported. Taken together this collection of data suggests that the abrogation of p53-dependent apoptosis may indeed impact upon carcinogenesis and neoplastic progression, but that the simplistic notion of p53 as the single gatekeeper of this pathway is untenable. PMID- 11024477 TI - Comet assay to sense neutron 'fingerprint'. AB - The suitability of comet assay to identify DNA damage induced by neutrons of varying energy was tested. For this purpose, monoenergetic neutrons from Hiroshima University Radiobiological Research Accelerator (HIRRAC) were used to induce DNA damage in irradiated human peripheral blood lymphocytes. The level of damage was computed as tail moment for different doses (0.125-1 Gy) and compared with the effects resulting from irradiation with (60)Co gamma. The neutron irradiated cells exhibited longer comet tails consisting of tiny pieces of broken DNA in contrast to the streaking tails generated by (60)Co gamma. The peak biological effectiveness occurred at 0.37 and 0.57 MeV; a further increase or decrease in neutron energy led to a reduced RBE value. The RBE values, as measured by the comet assay, were 6.3, 5.4, 4.7, 4.3, 2.6, and 1.7 for 0.37, 0.57, 0.79, 0.186, 1, and 2.3 MeV neutrons. The lower RBE value obtained by the comet assay when compared to that for other biological end points is discussed. This study reports the usefulness of the alkaline comet assay for identifying DNA damage induced by neutrons of the same radiation weighting factor. The comet assay is a potential tool for use in neutron therapy, as well as a method for the rapid screening of samples from individuals accidentally exposed to radiation. PMID- 11024476 TI - Hepatitis B virus-transfected Hep G2 cells demonstrate genetic alterations and de novo viral integration in cells replicating HBV. AB - Hepatitis B virus (HBV) is a major etiological factor associated with hepatocarcinogenesis, but its role in the transformation process remains unclear. We previously documented the accumulation of genetic alterations in a HBV transfected cell line. In the present study, we addressed the effect of HBV and its replication on the genome and phenotype of the host cell. Parental HBV-free Hep G2 cells and two HBV-transfected variant lines Hep G2215 and Hep G2T14. 1, which do and do not replicate HBV, respectively, were used to monitor genetic alterations in conjunction with HBV profile in vitro and in vivo. Comparison of in vitro growth rates showed that Hep G2T14.1 cells grew more rapidly, while Hep G2215 cells, replicating HBV, grew slower than parental Hep G2 cells. Molecular analysis confirmed an HBV integration site (s) in both variants, and reverse trancriptase-polymerase chain reaction (RT-PCR) amplification documented expression of transcript for the HBX protein, which has recently been implicated in the compromised efficiency of cellular DNA repair. Tumorigenisity testing indicate a comparable rate of tumor formation in nude mice of both HBV transfected variants, giving rise to tumors in 3 weeks; parental Hep G2 cells did not form tumors in nude mice. Tumor tissue from nude mice injected with Hep G2T14.1 cells showed no change in HBV status. However, a new HBV integration site was detected in tumor tissue from Hep G2215-injected mice. Two cell lines derived from the respective tumor tissue grew in vitro at rates compatible to those observe before passage in nude mice. The Hep G2215 tumor-derived line continued to replicate HBV, while HBV status remained unchanged in the Hep G2T14.1 tumor derived line. Unique genetic alterations were detected in both transfected cell lines, and Hep G2215 cells particularly showed cellular mosaicism and clonal selection when analyzed after the passage in nude mice. Further genetic alterations were detected in tumor-derived cell lines. Interestingly, the de novo genetic alterations in the Hep G2215 cells, which maintain the ability to replicate HBV, included a new HBV integration site, several chromosome rearrangements and loss of heterozygosity (LOH) of one p53 allele. Western analyses of p21/Waf1 protein indicate an upregulation of the protein in cells that replicate HBV. Based on the combined data, we hypothesize that the genetic alterations in the cellular genome could also be generated as a function in the presence of HBV and HBV replication. Possible mechanisms that could be implicated in cumulative mutagenetic events are discussed. PMID- 11024478 TI - Evidence that camptothecin-induced aberrations in the G(2) phase of cell cycle of Chinese hamster ovary (CHO) cell lines is associated with transcription. AB - It is widely accepted that camptothecin (CPT) is an S-dependent genotoxin. In this study, we aimed to elucidate the 'puzzling' induction of chromosomal damage by CPT in the G(2) phase of CHO cells, where no DNA synthesis is expected, focusing the attention on the possible role of the ongoing RNA synthesis, supposed to cause the conversion of CPT-single stranded cleavage complexes spaced closely on opposite DNA strands into DNA double strand breaks (DSB's) by the action of traversing RNA polymerase.CHO AA8 and its parental mutant EM9 cell lines were pre-treated with alpha-amanitin, which prevents transcription to pre-m RNA and challenged cells with CPT for the last hour in culture to evaluate whether G(2)-CPT-induced aberrations would have been reduced or abolished in the absence of RNA synthesis compared with G(2)-CPT treatment alone. The results obtained indicated a marked and significant reduction of aberration yields, to almost the control values (alpha-amanitin alone) when inhibition of RNA synthesis was substantial (3h total alpha-amanitin). Partial inhibition of RNA synthesis (2h total alpha-amanitin) slightly reduced the CPT-induced aberrations yield only at the high dose-level employed of CPT (20mM). This finding strongly supports the hypothesis that CPT-single stranded cleavages complexes spaced closely on opposite DNA strands are converted into DNA double strand breaks by the action of traversing RNA polymerase. PMID- 11024479 TI - Mutant frequencies and mutation spectra of dimethylnitrosamine (DMN) at the lacI and cII loci in the livers of Big Blue transgenic mice. AB - The lacI gene in Big Blue transgenic rodents has traditionally been used as a surrogate gene for in vivo mutations. Recently, a more efficient and less expensive assay involving direct selection in the smaller lambda cII gene has been developed. Little is known, however, about the comparative sensitivity of the two loci or their influence on the recovered mutation spectrum following mutagen treatment. We have compared the mutation frequency (MF) and mutational spectrum (MS) of lacI and cII from the same DNA samples isolated from the liver of control and dimethylnitrosamine (DMN)-treated mice. A three-fold (p<0.01) increase in the MF was observed at both loci in the DMN-treated group compared to the corresponding control groups. While the DMN-induced mutation spectrum at lacI was significantly different from its corresponding spontaneous mutation spectrum (p<0.001), the mutation spectrum at cII (p>0.28) was not. The mutation spectra at the two loci from the DMN-treated mice resembled each other but the 4, 2.5 and 12 fold increase in the mutation frequency of A:T>T:A transversions, single base deletions and deletions of more than four base pairs, respectively, at lacI, altered the spectra significantly (p<0.007). The number of mutations of these classes at cII was also increased, but the fractions were lower than at lacI. The spontaneous mutation spectra at the cII and lacI loci resembled each other except for the seven-fold increase in G:C or = 10 St II experiences irreversible conformational changes that notably reduce its biological activity. This reduced biological activity is associated with a partial defolding of the protein, which seems to contradict what is expected in terms of a molten globule formalism. PMID- 11024495 TI - Biochemical characterization of a thrombin-like enzyme and a fibrinolytic serine protease from snake (Agkistrodon saxatilis) venom. AB - A thrombin-like enzyme and a fibrinolytic serine protease were purified to homogeneity from the venom of a Korean snake Agkistrodon saxatilis emelianov. Both the purified enzymes migrated as a single protein band corresponding to 39 kDa in SDS-PAGE. However, the molecular mass was reduced to 28 kDa by enzymatic removal of the N-linked carbohydrates in those two different enzyme species. Although the thrombin-like enzyme and the fibrinolytic protease show homologous features in their molecular sizes and N-terminal amino acid sequences, yet they can be clearly distinguished from each other in terms of substrate specificity, susceptibility to inhibitors and fibrinogen degradation. It is postulated that these two enzymes are capable of functioning in a cooperative manner to effectively remove fibrinogen and consequently to reduce the blood viscosity. PMID- 11024496 TI - Determination of paralytic shellfish toxins in Portuguese shellfish by automated pre-column oxidation. AB - Automated pre-column oxidation (the method of Lawrence) was implemented on a routine basis since the end of 1996 to study paralytic shellfish poisoning (PSP) toxins in Portuguese shellfish. Liquid chromatography confirmed the presence of PSP toxins when the known toxic algae were present: Gymnodinium catenatum and/or Alexandrium cf. lusitanicum. On the other side, it has eliminated PSP toxins as a possible recurrent contaminant in oysters from Sado estuary. These oysters were already known to contain high levels of some metals (mainly zinc, copper and cadmium) due to their location in a contaminated area and their particular physiology prone to accumulate metals. The presence of PSP toxins in Scrobicularia plana from Mondego estuary and Tellina crassa from the northern coast, during the absence of the above toxic microalgae in the water column, was confirmed. Unlike other shellfish, these two genera have the feeding habit of aspirating more sediment than organisms in suspension, and probably ingest from the sediment resting cysts of PSP producing microalgae. This is another route of contamination that may help to explain why after a bloom certain shellfish species maintain toxicity for long periods. The method revealed to have a fast implementation on a daily basis, short analysis time (around 20 min between samples), high sensitivity and robustness, and therefore, it is one of the best HPLC methods for screening a large number of shellfish samples for monitoring purposes. PMID- 11024497 TI - Possible source of tetrodotoxin in the starfish Astropecten scoparius. AB - The seasonal variations of toxicity and stomach contents in toxic starfish Astropecten scoparius were detected. The average highest specimen toxicity, expressed as tetrodotoxin (TTX), was 16,821 mouse units (MU). The toxin was composed of TTX only, except April's sample containing mainly TTX along with minor paralytic shellfish poisons. The composition in the stomach of less and more toxic starfish was mainly Veremolpa scabra and Umborium suturale, respectively. The toxicities of U. suturale and V. scabra, found in the starfish stomach were 65 and 33 MU, respectively. For those collected from the coastal waters, U. suturale was toxic with average toxicity value of 77 MU/g, but V. scabra was nontoxic. The toxin in the specimens of U. suturale collected from either the digestive gland of starfish or the coastal waters, was TTX and anhydroTTX only. It indicates that the starfish A. scoparius might mainly accumulate high amount of TTX from U. suturale. Furthermore, both small gastropods U. suturale and Natica psuestes are first reported to contain TTX. PMID- 11024498 TI - Effects of brucine, a plant alkaloid, on M(1) muscarinic receptors and alpha(1) adrenoceptors in the rabbit vas deferens preparation. AB - The plant alkaloid brucine is an analogue of strychnine and is known to be an allosteric modulator at cloned M(1) muscarinic receptors. The functional effects of brucine were examined on the M(1) muscarinic receptors in the rabbit isolated vas deferens preparation. Brucine (10-100 microM) enhanced the effects of the muscarinic agonist McN-A-343 at presynaptic M(1) muscarinic receptors in the rabbit isolated vas deferens preparation, but only when brucine was added prior to McN-A-343. This effect is indicative of a positive allosteric action. It was poorly reversed on washing. Brucine did not affect the responses to the mamba venom muscarinic toxins MT2 and MT4, which are also allosteric activators in this preparation. Brucine (10-100 microM) caused a significant decrease in the twitch response to electrical stimulation in the rabbit vas deferens preparation, which was not antagonised by 100 nM pirenzepine (an M(1) muscarinic antagonist). Brucine and MT4, but not MT2, caused significant decreases (p<0.05) in the responses to noradrenaline in the rabbit vas deferens preparation. Responses to ATP and KCl were not affected. In radioligand binding assays, brucine displaced the alpha(1)-adrenoceptor ligand prazosin from its specific binding sites in membranes made from rat cerebral cortex and rat vas deferens. The apparent K(i) values were 150 and 3.4 microM in the cortical and vas deferens membranes, respectively. The positive allosterism found with brucine at cloned M(1) receptors seems to be mirrored at native M(1) receptors. However, the unexpected blocking effects at alpha(1)-adrenoceptors indicates that more selective ligands than brucine are required as starting points for the design of specific enhancers of the activity of M(1) receptors with therapeutic potential. PMID- 11024499 TI - Losses of the cyanobacterial toxin microcystin-LR from aqueous solution by adsorption during laboratory manipulations. AB - The effect of plastic and methanol on the loss of microcystin-LR from solution was analysed by HPLC with photodiode array detection (HPLC-PDA). With plastic disposable pipette tips, the loss from an aqueous microcystin-LR (MC-LR) solution was 4.2% per tip operation. Using the same pipette tip, four operations were required to completely saturate a single tip with toxin. MC-LR attached to plastic pipette tips could subsequently be eluted by methanol and detected by HPLC-PDA. At methanol concentrations below 25% (v/v), recovered concentrations of MC-LR decreased significantly. Differences in MC-LR concentration were also noted by performing 50% dilution with Milli-Q water or methanol. The results are discussed in relation to the hydrophobicity of MC-LR, analytical procedures and the avoidance of toxin losses from solution during laboratory manipulations. PMID- 11024500 TI - In this issueellipsis PMID- 11024501 TI - Gene therapy for brain tumors: the fundamentals. AB - BACKGROUND: Over the past two decades, significant advances have been made in the fields of virology and molecular biology, and in understanding the genetic alterations present in brain tumors. The knowledge gained has been exploited for use in gene therapy. OBJECTIVE: The purpose of this article is to present an introduction to the field of brain tumor gene therapy for the practicing clinician. RESULTS: A variety of gene therapy strategies have now been used in the laboratory and in clinical trials for brain tumors. They can be divided into five categories: 1) gene-directed enzyme prodrug ("suicide gene") therapy (GDEPT); 2) gene therapy designed to boost the activity of the immune system against cancer cells; 3) oncolytic virus therapy; 4) transfer of potentially therapeutic genes--such as tumor suppressor genes--into cancer cells; and 5) antisense therapy. GDEPT is the strategy that has been most extensively studied. CONCLUSIONS: To date, gene therapy has been found to be reasonably safe and concerns related to adverse events such as insertional mutagenesis have not been realized. Although patients have not been cured, the development of this therapy could still be considered to be at an early stage. Current research is addressing factors that could be limiting the successful clinical application of gene therapy, which remains an intriguing experimental option for patients with malignant brain tumors. PMID- 11024502 TI - A limited anterior petrosectomy with preoperative embolization of the inferior petrosal sinus for ventral brainstem tumor removal. AB - BACKGROUND: The present study describes the use of a limited subtemporal extradural anterior petrosectomy with preoperative embolization of the inferior petrosal sinus for the management of tumors located behind the clivus and ventral to the brainstem. Details of the procedure and its application in five cases are presented. METHODS: This procedure consists of using the extradural route to approach the upper side of the petrosal pyramid so that it can be drilled medially, and to resect the apex to come out into the posterior fossa. This route gives a petrosectomy just medial to the horizontal segment of the petrous carotid artery in front of the cochlea. It goes around the labyrinthine mass and the internal auditory canal from above to expose the posterior fossa dura between the two petrosal sinuses. The dural opening exposes the ventral aspect of the pons from the trigeminal nerve to the origin of the abducens nerve, ventral to the facial nerve. Preoperative embolization of the inferior petrosal sinus allows its intraoperative section for a wider exposure along the lower clivus. This approach can easily be combined with an intradural approach to provide additional exposure above the trigeminal nerve. Patients who underwent this procedure had prepontine cisternal chordoma or epidermoid cyst of the petroclival region. RESULTS: One patient experienced a cranial nerve deficit as a direct result of the surgical procedure (VIth nerve palsy requiring surgery) but no other patient has had permanent neuromuscular compromise. Complications consisted of a wound infection in one case. Tumor removal was total in three cases and partial in two cases. CONCLUSION: Quite easy to master, the anterior petrosectomy with preoperative embolization of the inferior petrosal sinus is a time-conserving approach giving one of the best routes to reach the ventral brainstem while working in front of the cranial nerves and preserving hearing. PMID- 11024503 TI - Combined treatment of fourth ventricle ependymomas: report of 26 cases. AB - BACKGROUND: This study investigated the relevance of prognostic factors and the impact of histological features in posterior fossa ependymoma. METHODS: The charts of 26 patients (aged 1-59 years, mean 20.6 years; 11 adults) with posterior fossa ependymoma operated on between January 1983 and December 1994 were reviewed and patients followed up (mean: 93 months). RESULTS: Gross total resection was performed in 18 patients (69%), subtotal in seven patients (27%), biopsy in one patient (4%). One patient (3.8%) developed respiratory complications and died. All patients underwent posterior fossa radiotherapy (5000 cGy) after surgery. Four children first received chemotherapy and then radiotherapy only when at least 3 years old. Eleven patients (42%) received radiotherapy and subsequently chemotherapy. The 5-year survival rate was 90% for adults and 40% for children ( 2 nkat g dry biomass(-)(1) were isolated. Among them three new D-amino acid oxidase producers were found and identified as Fusarium oxysporum, Verticilium lutealbum and Candida parapsilosis. The best new D-amino oxidase producer was the fungus F. oxysporum with a D-amino acid oxidase activity of about 900 nkat g dry biomass( )(1) or 21 nkat mg protein(-)(1). With regard to the use as a biocatalytic tool in biotechnology the substrate specificities of the three new D-amino acid oxidases were compared with those of the known D-amino acid oxidases from Trigonopsis variabilis, Rhodotorula gracilis and pig kidney under the same conditions. All six D-amino acid oxidases accepted the D-enantiomers of alanine, valine, leucine, proline, phenylalanine, serine and glutamine as substrates and, except for the D-amino acid oxidase from V. luteoalbum, D-tryptophane, D tyrosine, D-arginine and D-histidine were accepted as well. The relative highest activities (>95%) were measured versus D-alanine (C. parapsilosis, F. oxysporum, T. variabilis), D-methionine (V. luteoalbum, R. gracilis), D-valine (T. variabilis, R. gracilis) and D-proline (pig kidney). The D-amino oxidases from F. oxysporum and V. luteoalbum were able to react with the industrially important substrate cephalosporin C although the D-amino acid oxidase from T. variabilis was at least about 20-fold more active with this substrate.As the results of our studies, a reliable oxidase assay was developed, allowing high throughput screening in a microplate reader. Furthermore, three new microbial D-amino acid oxidase-producers with interesting broad substrate specificities were introduced in the field of biotechnology. PMID- 11024525 TI - Kinetics of thermal deactivation of enzymes: a simple three parameters phenomenological model can describe the decay of enzyme activity, irrespectively of the mechanism. AB - Heat induced enzyme inactivation or protein denaturation is now well documented, due to progresses in methods, instruments and computation resources. Complex mechanisms, rather than the classic simple "one step - two states" model (still in use) are recognized in many cases, leading investigators to manipulate more or less complicated kinetic expressions describing the heat induced decay of enzyme activity.We show that the different kinetic expressions related to different mechanisms among the most frequently encountered can be arranged in a common simple three-parameters biexponential equation.This unifying simplification is of interest for people focusing attention to phenomenological rather than mechanistic description of the kinetics of heat induced enzyme deactivation. Moreover, the reduction in the number of parameters reduces the risk of cross correlation and allows a better estimation of the apparent rate constants (which are in many cases the pertinent required information). It also illustrates the difficulty to make inference of mechanism from kinetics, since the same equation applies for a variety of mechanisms ("kinetic homeomorphism") - in particular, it stresses out the need of caution when reporting on existence of isoenzymes from deactivation kinetics.Application of this simple 3-parameters biexponential kinetic expression has been validated with a number of results in the Literature and current investigations in our laboratory. Two examples are given. PMID- 11024526 TI - Kinetic analysis of red pigment and citrinin production by Monascus ruber as a function of organic acid accumulation. AB - In submerged cultures performed in synthetic medium containing glucose and glutamate, the filamentous fungus Monascus ruber produced a red pigment and a mycotoxin, citrinin. In oxygen-limiting conditions, the production of these two metabolites was growth-associated, as was the production of primary metabolites. In oxygen-excess conditions, the profile of citrinin production was typical of a secondary metabolite, since it was produced mostly during the stationary phase. In contrast, the production of the pigment decreased rapidly throughout the culture, showing a profile characteristic of an inhibitory mechanism. The organic acids produced during the culture, L-malate and succinate, were shown to be slightly inhibitory against pigment production, while citrinin production was unaffected. However, this inhibition could not account for the observed profile of pigment production in batch cultures. Other dicarboxylic acids such as fumarate or tartrate showed a similar effect to that provoked by malate and succinate as regards pigment production. It was concluded that the decrease in red pigment production during the culture was due to the inhibitory effect of an unknown product whose accumulation was favored in aerobic conditions. PMID- 11024527 TI - Mycelium-bound carboxylesterase from Aspergillus oryzae: an efficient catalyst for acetylation in organic solvent. AB - Dry mycelium of a strain of Aspergillus oryzae efficiently catalyzed the esterification between free acetic acid and primary alcohols (geraniol and ethanol) in organic solvent. The growth conditions to obtain high activity of mycelium-bound enzymes were firstly evaluated. A medium containing Tween 80 as carbon source furnished mycelium with the highest activity in the hydrolysis of alpha-naphthyl esters (alpha-N-acetate, butyrate, caprylate). Dry mycelium was employed to select suited conditions for an efficient acetylation of ethanol and geraniol in heptane. Maximum productions were obtained using 30 g l(-)(1) of lyophilized cells: 12.4 g l(-)(1) of geranyl acetate were produced at 80 degrees C starting from 75 mM geraniol and acetic acid (84% molar conversion) and 4.1 g l(-)(1) of ethyl acetate at 50 degrees C from 50 mM ethanol and acetic acid (94% molar conversion) after 24 h. The stability of the mycelium-bound carboxylesterases are notable since only 10-30% loss of activity was observed after 14 days at temperatures between 30 and 50 degrees C. PMID- 11024528 TI - Increase in Chlorella strains calorific values when grown in low nitrogen medium. AB - The calorific value of five strains of Chlorella grown in Watanabe and low nitrogen medium was determined. The algae were grown in small (2L) stirred tank bioreactors and the best growth was obtained with Chlorella vulgaris with a growth rate of 0.99 d(-1) and the highest calorific value (29 KJ/g) was obtained with C. emersonii. The cellular components were assayed at the end of the growth period and the calorific value appears to be linked to the lipid content rather than any other component. PMID- 11024529 TI - Characterization of T cell receptor (TCR) of organ-specific autoimmune disease inducing T cells and TCR-based immunotherapy with DNA vaccines. AB - Organ-specific autoimmune diseases and their animal models are characterized by the finding that the development of the diseases is closely associated with, or induced by, T cells reactive to organ-specific antigens. Therefore, the identification of T cell receptors (TCR) used by disease-inducing T cells within a short period of time is a key factor for designing TCR-based immunotherapy. The findings introduced in this article show that TCR associated with the development of multiple sclerosis and experimental autoimmune diseases including encephalomyelitis (EAE), neuritis (EAN) and carditis (EAC) are identifiable by complementarity-determining region 3 (CDR3) spectratyping analysis and subsequent sequencing of the CDR3 region of spectratype-derived TCR clones. It is also demonstrated that immunotherapy targeting disease-associated TCR using monoclonal antibodies and DNA vaccines significantly reduced the histological severity, and completely suppressed the inflammation in some animals. Since depletion or suppression of one of several types of effector cells does not significantly improve the severity of the disease, combined TCR-based immunotherapy should be considered as a primary therapy for T cell-mediated autoimmune diseases. TCR based immunotherapy after rapid identification of autoimmune disease-associated TCR by CDR3 spectratyping can be applicable, not only to animal, but also to human autoimmune diseases whose pathomechanism is poorly understood. PMID- 11024530 TI - Amelioration of CR-EAE with lisofylline: effects on mRNA levels of IL-12 and IFN gamma in the CNS. AB - CR-EAE is a Th1-mediated inflammatory autoimmune demyelinating disease of the CNS and serves as a model of human multiple sclerosis. Our previous studies have shown the protective effect of orally administered lisofylline in the prevention of active and passively induced acute EAE. In our present studies we have examined the efficacy and mechanism of action of lisofylline on CR-EAE. Lisofylline decreased the number and severity of paralytic attacks in mice with relapsing EAE. The reduction of clinical disease correlated with decreased levels of mRNA levels of IFN-gamma but not of mRNA levels of IL-12. These studies suggest that lisofylline may be an effective therapeutic for established Th1 mediated autoimmune disease and that it acts by blocking IL-12R signaling and not IL-12 production in vivo. PMID- 11024531 TI - Interleukin-1beta, -6, -12 and tumor necrosis factor-alpha expression in brains of dogs with canine distemper virus infection. AB - Canine distemper virus infection in dogs is commonly associated with demyelinating central nervous system lesions. Investigations on viral protein expression by studying mRNA and protein distribution together with the characterization of CD4 and CD8 inflammatory cells and MHC class II up-regulation revealed a biphasic disease process. To further investigate the cellular interactions in the different plaque types the cerebella of 14 dogs with confirmed distemper infection were investigated for expression of interleukin (IL)-1beta, -6, -12 and tumor necrosis factor-alpha (TNF) by immunohistochemistry using rabbit polyclonal anti-cytokine antibodies. T-cells and astrocytes were identified with rabbit anti CD3- and GFAP-monoclonal and polyclonal antibodies, respectively; and microglia/macrophages were characterized by their ability to bind lectin from Bandeiraea simplicifolia (BS-1). To further name the cytokine expressing cells immunoenzymatic double staining using DAB and New Fuchsin was performed. White matter lesions were classified according to histopathological criteria into acute, subacute and chronic. Canine distemper virus nucleoprotein antigen was demonstrated in nearly all plaques, except in older plaques where virus was not present within the plaque but adjacent to the lesion. IL-1 expression was observed to varying degrees in all types of lesions. Most often IL 1 was present in CD3 and BS-1 positive cells in the brain parenchyma in earlier plaques and comprising perivascular cuffs found in chronic plaques. IL-6 expression was present in all lesions, and followed a similar distribution pattern as IL-1. IL-12 displayed very often a granular extracellular pattern of immunoreactivity, especially in the brain parenchyma, and was found only in individual perivascular cells. TNF staining, predominantly found in astrocytes, was present in lesions of various types; however, staining appeared to be stronger in acute lesions and decreased in chronic plaques. In the latter, TNF seemed to be more prominent in areas adjacent to the plaques. Summarizing, in early non-demyelinating lesions without overt inflammation TNF seemed to be important, whereas in distemper lesions with inflammatory infiltrates IL-1 and to a lesser degree IL-6 were more prominent. These results imply that TNF may be involved in the pathogenesis of early demyelination in nervous distemper. PMID- 11024532 TI - Delayed onset and decreased severity of experimental autoimmune uveoretinitis in mice lacking nitric oxide synthase type 2. AB - To investigate the role of nitric oxide (NO), produced by the inducible form of NO synthase (NOS-2) in the development of experimental autoimmune uveoretinitis (EAU), we immunized C57BL/6x129Sv (H-2(b)) mice carrying a targeted disruption of the gene encoding NOS-2 (NOS-2[-/-]), and wild-type (WT) C57BL/6x129Sv controls with interphotoreceptor retinoid binding protein (IRBP). NOS-2[-/-] mice developed a clinical EAU with delayed onset and decreased severity compared to WT controls. The ocular tissues from WT mice contained activated F4/80 macrophages with NOS-2 expression and retinal destruction whereas less intense EAU was detected in NOS-2[-/-] mice. The expression of NOS-2 mRNA was detected in the retina at the peak of EAU in WT. Analysis of cytokine production in the spleen from NOS-2[-/-] mice by RT-PCR showed high levels of IL-10 mRNA. Our results demonstrate that NO is clearly involved in EAU and may be important for the regulation of immune responses through the regulation of IL-10. PMID- 11024533 TI - Mechanisms underlying chemical sympathectomy-induced suppression of herpes simplex virus-specific cytotoxic T lymphocyte activation and function. AB - Lymphoid tissues are extensively innervated by noradrenergic fibers of the sympathetic nervous system. 6-hydroxydopamine (6-OHDA)-induced chemical sympathectomy is commonly used to assess the impact of this innervation on immune function. Using the glucocorticoid receptor antagonist RU486, the mineralocorticoid receptor antagonist spironolactone, and the beta-adrenergic receptor antagonist nadolol, the roles of corticosterone and norepinephrine in sympathectomy-mediated modulation of both the primary and memory cellular immune responses to herpes simplex virus type 1 (HSV-1) infection was investigated. We demonstrated that both of these immunomodulators play a role in mediating sympathectomy-induced suppression of the generation of HSV-specific primary cytotoxic T lymphocytes (CTL) and the activation of HSV-specific memory CTL (CTLm). Furthermore, we demonstrated a role for both Type I and Type II corticosteroid receptors in the regulation of HSV-specific immunity. Overall, these findings not only further support a role for neuroendocrine-mediated modulation of immune function, but also a need to exercise caution in attributing the effects of chemical sympathectomy to solely the absence of sympathetic innervation of lymphoid tissues. PMID- 11024534 TI - Altered beta-adrenoceptor function associated to protein kinase C activation in hyperproliferative T lymphocytes. AB - beta-Adrenoceptor (betaAR) expression and function as well as its modulation via intracellular transduction signals, were analyzed on the T cell lymphoma BW5147. Independently to the kinetic of proliferation and relative to the number of receptors displayed in normal T lymphocytes, BW5147 cells displayed a decreased number of betaAR, uncoupled to adenylate cyclase, but coupled to protein kinase C stimulation. This last effect was impaired by a beta-antagonist and by blockers of the enzymatic pathways involved in T lymphocyte proliferation, inducing a recovery of betaAR sites. Down-regulation of betaAR would implicate the loss of a negative neuroimmune control mechanism for lymphocyte proliferation. The coupling of the remaining sites to a positive signal for cellular activation, would contribute to establish an hyperproliferative state. PMID- 11024535 TI - Evidence for an interferon-related inflammatory reaction in the trisomy 16 mouse brain leading to caspase-1-mediated neuronal apoptosis. AB - The trisomy of human chromosome 21 (Down syndrome) is the leading genetic cause of learning difficulties in children, and predisposes this population to the early onset of the neurodegeneration of Alzheimer's disease. Down syndrome is associated with increased interferon (IFN) sensitivity resulting in unexpectedly high levels of IFN inducible gene products including Fas, complement factor C3, and neuronal HLA I which could result in a damaging inflammatory reaction in the brain. Consistent with this possibility, we report here that the trisomy 16 mouse fetus has significantly increased whole brain IFN-gamma and Fas receptor immunoreactivity and that cultured whole brain trisomy 16 mouse neurons have increased basal levels of caspase 1 activity and altered homeostasis of intracellular calcium and pH. The trisomic neurons also showed a heightened sensitivity to the increase in both Fas receptor levels and caspase 1 activity we observed when IFN-gamma was added to the neuron culture media. Because of the autoregulatory nature of IFN activity, and the IFN inducing capability of caspase 1-activated cytokine activity, our data argue in favor of the possibility of an interferon-mediated, self-perpetuating, inflammatory response in the trisomy brain that could subserve the loss of neuron viability seen in this trisomy 16 mouse model for Down syndrome. PMID- 11024536 TI - Astrocytes and central nervous system endothelial cells do not express B7-1 (CD80) or B7-2 (CD86) immunoreactivity during experimental autoimmune encephalomyelitis. AB - The identity of cell types within the central nervous system (CNS) capable of activating T lymphocytes is a fundamental issue in the understanding of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). To become fully activated, a T cell must recognize its antigen and receive co stimulation, the latter being optimally delivered via B7-1 and/or B7-2 molecules expressed by the antigen presenting cell (APC). There are conflicting reports regarding whether astrocytes or CNS endothelial cells (EC) can act as fully competent APCs. The present studies were performed to determine whether astrocytes or CNS EC express B7-1 or B7-2 immunoreactivity during EAE. No expression of B7-1 or B7-2 by either astrocytes or EC was detected during acute, remitting, relapsing or chronic EAE, whether EAE was induced by active immunization or cell transfer using five different myelin antigens. These results suggest that neither astrocytes nor CNS EC can deliver co-stimulatory signals via B7 molecules in the setting of murine EAE, rendering them incapable of acting as fully competent APCs. PMID- 11024537 TI - Rhesus monkeys are highly susceptible to experimental autoimmune encephalomyelitis induced by myelin oligodendrocyte glycoprotein: characterisation of immunodominant T- and B-cell epitopes. AB - Eight rhesus monkeys with different MHC backgrounds were immunized with myelin oligodendrocyte glycoprotein (MOG). All developed severe experimental autoimmune encephalomyelitis associated with large inflammatory foci and extensive demyelination. T-cell autoreactivity to MOG was directed against three main epitopes encompassed within amino acids 4-20, 35-50 and 94-116, of which two are also immunodominant epitopes for the autoimmune T cell response to MOG in patients with MS. A strong B cell response to MOG was observed in all monkeys and major epitopes recognized were located within amino acids 4-26, 24-46 and 44 66/54-76. PMID- 11024538 TI - Vasoactive intestinal peptide and pituitary adenylate cyclase activating polypeptide inhibit the MEKK1/MEK4/JNK signaling pathway in LPS-stimulated macrophages. AB - The vasoactive intestinal peptide (VIP) and the pituitary adenylate cyclase activating polypeptide (PACAP), two immunomodulatory neuropeptides that affect both innate and acquired immunity, downregulate TNFalpha expression in LPS stimulated peritoneal macrophages and Raw 264.7 cells. We showed previously that VIP/PACAP change the composition of the CRE-binding complex in the TNFalpha promoter from highc-Jun/(low)CREB, characteristic for LPS-stimulated macrophages, to lowc-Jun/(high)CREB, characteristic for the unstimulated cells. In the present study we examined the effects of VIP/PACAP on the MEKK1/MEK4/JNK transduction pathway, and on the subsequent changes in Jun family members. Our studies indicate that VIP/PACAP inhibit MEKK1 activity, and the subsequent phosphorylation of MEK4, JNK, and c-Jun. Treatment with VIP or PACAP results in a decrease in AP-1 binding, and a marked change in the composition of the AP-1 complexes from c-Jun/c-Fos to JunB/c-Fos. Western blots confirm that VIP stimulates JunB production in LPS-stimulated macrophages. Both the inhibition of the MEKK1/MEK4/JNK pathway, leading to the reduction in phosphorylated c-Jun, and the stimulation of JunB, are mediated through the specific VPAC1 receptor and the cAMP/PKA pathway. The VIP/PACAP interference with the stress-induced SAPK/JNK pathway in stimulated macrophages may represent a significant element in the regulation of the inflammatory response by the endogenous neuropeptides. PMID- 11024540 TI - Heme oxygenase-1 in lesions of rat experimental autoimmune encephalomyelitis and neuritis. AB - The enzyme heme oxygenase-1 (HO-1) is reducing heme to the gaseous mediator carbon monoxide, to iron and the antioxidant biliverdin. The inducible expression of HO-1 is considered a protective cellular mechanism against reactive oxygen intermediates. Further, carbon monoxide (CO) is a regulator of cGMP synthesis, of NO-synthetases and cyclooxygenases, thereby indirectly modulating reactive processes. Here we report expression of HO-1 in rat experimental autoimmune encephalomyelitis (EAE) and neuritis (EAN). With both models, similar results were obtained: HO-1 was localized predominantly to infiltrating, monocytic, but only rarely to ramified microglial cells or astrocytes surrounding the inflammatory lesions. Prominent expression by monocytic cells was seen from day 11 after immunization correlating with the development of neurologic disease. Further, local expression is persistent for long after cessation of neurologic signs. Thus, HO-1 could be considered a factor in the formation and resolution of inflammatory autoimmune lesions of the nervous system. PMID- 11024539 TI - A pathogenic role of Th2 responses on the severity of encephalomyelitis induced in mice by herpes simplex virus type 2 infection. AB - A pathogenic role of Th2 cells and their cytokine products (IL-4 and IL-10, Th2 cytokines) on the development of herpes simplex myelitis (HSM) was studied in mice exposed to footpad injection of herpes simplex virus type 2 (HSV-2). Morbidity and mortality of mice with HSM (HSM mice) increased when they were treated with a mixture of Th2 cytokines. Additionally, survival rates of HSM mice increased when they were treated with a mixture of mAbs for Th2 cytokines. As compared with HSM mice treated with saline, the growth of HSV-2 in spinal cords of HSM mice treated with the mixture of Th2 cytokines increased. Th2 cells (myelitis-associated Th2 cells, MTh2 cells) were demonstrated among cerebrospinal fluid cells from HSM mice. After the stimulation with HSV-2 antigen (Ag), MTh2 cells from HSM mice previously treated with the mixture of Th2 cytokines produced enhanced amounts of Th2 cytokines into their culture fluids, as compared with the amount of Th2 cytokines produced by MTh2 cells. Th2 cells were also demonstrated in mononuclear cells from spleens of HSM mice. When compared with HSM mice inoculated with splenic CD4(+) T cells from normal mice, morbidity and mortality of HSM mice inoculated with MTh2 cells markedly increased. These results indicated that the severity of HSM induced in mice by footpad injection of HSV-2 was influenced by MTh2 cells or Th2 cytokines released from these MTh2 cells. Th2 responses manifested in mice by HSV-2 infection may act as a pathogenic enhancer of HSM severities. PMID- 11024541 TI - Sequential expression of chemokines in experimental autoimmune neuritis. AB - Recruitment of inflammatory cells is of critical importance in the pathogenesis of immune-mediated demyelinating diseases in the peripheral nervous system (PNS). Evidence is increasing that chemokines might play a key role in this process, since they promote leukocyte entry into the nervous system during immune-mediated inflammation. In the present study we report the expression pattern of the chemokines interferon-gamma-inducible protein (IP)-10, monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and regulated upon activation normal T cell expressed and secreted (RANTES) in sciatic nerves from animals with myelin-induced experimental autoimmune neuritis, using a semiquantitative reverse transcriptase-PCR dot-blot hybridization assay. The mRNAs for MIP-1alpha and MIP-1beta were found to be upregulated with peak values at day 13 post-immunization (p.i.), preceding maximum disease severity. In contrast, mRNAs for MCP-1, RANTES, and IP-10 exhibited peak levels coincident with peak of the disease at day 15 p.i. Increased mRNA expression was associated with enhanced protein levels, as demonstrated by immunoblotting for each chemokine investigated. Immunohistochemistry for IP-10 protein revealed immunoreactivity associated with perineurial endothelial cells. RANTES protein was localized immunohistologically to invading T lymphocytes. Our findings suggest that chemokines, which act towards T cells and mononuclear phagocytes, are sequentially upregulated during the clinical course of EAN and thus may contribute to the pathogenesis of inflammatory demyelinating diseases of the PNS. PMID- 11024542 TI - Presumed immune-mediated cerebellar granuloprival degeneration in the Coton de Tulear breed. AB - An unusual form of cerebellar granuloprival degeneration was observed in three male Coton de Tulear puppies between 12 and 14 weeks of age from different litters showing progressive cerebellar signs beginning at 8 weeks after birth. Pathological examinations revealed a shrunken cerebellum. Histopathologically the granular cells were diminished or almost completely absent, some 'torpedos' of Purkinje cells were present. There was a marked gliosis, and occasionally small inflammatory foci were present. A marked diffuse T cell infiltration (CD3(+) cells) occurred in the lesions, B cells did not appear. CD18 staining showed an upregulation of microglial cells at the lesion site. Histopathologically the lesions resembled paraneoplastic cerebellar degeneration which is caused by an autoimmune mediated T cell reaction. This congenital condition in the Coton de Tulear dog breed could be based on a genetically defined immune defect leading to autoimmune destruction of the granular cells. PMID- 11024543 TI - Interleukin 1 receptor accessory protein (IL-1RAcP) is necessary for centrally mediated neuroendocrine and immune responses to IL-1beta. AB - Mice deficient for the IL-1RAcP gene (IL-1RAcP KO) were used to explore the role of IL-1RAcP in physiological functions of brain IL-1beta. Animals were injected i.c.v. with two different doses of recombinant human (rh) IL-1beta: a small one (750 pg) known to induce sickness behavior, and a larger one (50 ng), chosen to counteract the possible loss of affinity of IL-1beta on its receptor. Neuroendocrine and immune parameters were measured 2 h after IL-1 injection. The increase of plasma corticosterone induced by rhIL-1beta in wild-type (WT) mice was not observed in IL-1RAcP KO mice. Likewise, the depression of splenocyte proliferation occurred in WT but not in KO mice. Finally, in opposition to WT mice, plasma levels and brain cortical content of IL-6 in IL-1RAcP KO mice remained unchanged as compared to saline-injected controls. The results clearly demonstrate that IL-1RAcP is necessary for the induction of the main neuroendocrine and immune effects of central IL-1beta. PMID- 11024544 TI - Aminoguanidine reduces apoptosis of circulating V Beta 8.2 T lymphocytes in Lewis rats with actively induced experimental autoimmune encephalomyelitis. Association with persistent inflammation of the central nervous system and lack of recovery. AB - Aminoguanidine therapy delayed the onset of actively induced EAE in Lewis rats, but recovery was impaired in most animals. In the central nervous system this was correlated with persistent inflammation and production of proinflammatory cytokines. In the periphery of aminoguanidine-treated animals, T lymphocytes showed increased proliferation against myelin basic protein, and the percentage of Vbeta 8.2(+) T lymphocytes undergoing early apoptosis was markedly decreased, although it was unchanged in Vbeta 8.2(+) T cells isolated from the spinal cord. These results suggest that the prolonged survival of circulating encephalitogenic cells achieved by aminoguanidine would favor a longer lasting entry of these cells into the nervous system resulting in persistent inflammation and lack of recovery. PMID- 11024545 TI - Characterization and visualization of [125I] stromal cell-derived factor-1alpha binding to CXCR4 receptors in rat brain and human neuroblastoma cells. AB - Stromal cell-Derived Factor-1 (SDF-1alpha), binds to the seven-transmembrane G protein-coupled CXCR4 receptor and modulates cell migration, differentiation, and proliferation. CXCR4 has been reported to be expressed in various tissues including brain. Moreover, CXCR4 has recently been shown to be one of the coreceptors for HIV-1 infection which could be implicated in HIV encephalitis. In the present study, the binding properties and autoradiographic distribution of [125I]SDF-1alpha binding to CXCR4 were characterized in the adult rat brain. SDF 1alpha binding and CXCR4 coupling system were also studied in human neuroblastoma cell line SK-N-SH. The binding of [125I]SDF-1alpha on rat brain sections was specific, time-dependent and reversible. The highest densities of CXCR4 were detected in the choroid plexus of the lateral and the dorsal third ventricle. Lower densities of [125I]SDF-1alpha binding sites were observed in various brain regions including cerebral cortex, anterior olfactory nuclei, hippocampal formation, thalamic nuclei, blood vessels and pituitary gland. In the choroid plexus, the IC(50) and K(d) of [125I]SDF-1alpha binding were respectively 0.6 nM and 0. 36 nM. Similar IC(50) values were obtained in other brain structures. A CXCR4 antagonist, bicyclam, competed with SDF-1alpha binding (30% inhibition at 10(-6) M). In SK-N-SH cells, [125I]SDF-1alpha bound to CXCR4 with a K(d) of 5.0 nM and a maximal binding capacity of 460 fmol/mg of protein. SDF-1alpha induced a rapid and transient intracellular calcium increase in SK-N-SH cells. These findings suggest that CXCR4 is highly expressed in some brain structures and have a regulatory role in the nervous system. The significance of this expression in the brain parenchyma and more specifically in the choroid plexus remains to be clarified in the normal as well as in the infected brain. PMID- 11024546 TI - Anti-CD40, anti-CD3, and IL-2 stimulation induce contrasting changes in CB1 mRNA expression in mouse splenocytes. AB - The expression and function of cannabinoid receptor 1 (CB1) in mouse immune cells is unclear. Here we show that splenic B cells express more CB1 mRNA than T cells. Furthermore, splenocytes stimulated with the T cell mitogens, PMA/Io and anti CD3, showed a decrease in CB1 message while cultures stimulated with the B cell mitogen, anti-CD40 antibody, showed an increase in message. In addition, co treatment with mitogens and IL-2 uniformly caused an increase in CB1 mRNA. It is suggested that signaling pathways activated by T cell mitogens lead to decreased CB1 gene activation while pathways activated by B cell mitogens and IL-2 lead to increased CB1. PMID- 11024547 TI - Genetic control of pathogenic mechanisms in autoimmune demyelinating disease. AB - Multiple sclerosis is a disease of discrete phenotypes in different individuals. Animal models have been useful in identifying self-antigens that become the focus of autoimmune attack and genetic loci that control susceptibility to disease. We have previously demonstrated a role for Fas-dependent pathogenesis in the induction of EAE in B10.PL mice immunized with MBP. Others have indicated a Fas independent mechanism predominates in SJL mice immunized with PLP. Here we compare the response of (B10.PLxSJL)F1 and parental mice under similar conditions for induction of EAE. The results indicate that immunodominance and dominant pathogenic mechanisms are both under genetic control, but can be inherited independently. The data also indicate that the dominant pathogenic mechanism can change during the course of disease in an individual. Elucidation of the genetic elements controlling pathogenesis during the course of disease would provide important information in designing therapeutic strategies for individuals in a heterogeneous patient population. PMID- 11024548 TI - The chemokine receptor CCR2 is involved in macrophage recruitment to the injured peripheral nervous system. AB - Wallerian degeneration is one of the most elementary reactions of the nervous system after transection of axons, leading to the recruitment of mononuclear cells from the systemic circulation. However, the exact mechanisms regulating this cell invasion have not yet been clarified in detail. Chemokines and their receptors play a central role in leukocyte trafficking, in particular the chemokine MCP-1 has been strongly implicated in macrophage recruitment to the injured nervous system. The present study investigates the course of Wallerian degeneration after transection of the sciatic nerve in mice deficient in two chemokine receptors: CCR2, the main receptor for MCP-1, and CCR5, a marker for Th1 T lymphocytes but also present on macrophages. The number of invading macrophages was determined by immunocytochemistry for three typical macrophage antigens (F4/80, Mac-1, LFA-1). The chemokine receptor CCR2 was expressed by infiltrating cells in the transected nerve stumps. Macrophage invasion was significantly impaired in CCR2-knockout mice when compared with wildtype controls and CCR5-deficient mice. Subsequently, there was a corresponding decrease in myelin phagocytosis due to the reduced invasion of phagocytic macrophages. These data demonstrate the involvement of the chemokine receptor CCR2 in macrophage recruitment to the injured nervous system. PMID- 11024549 TI - TNF-receptor 1 deficiency fails to alter the clinical and pathological course in mice with globoid cell leukodystrophy (twitcher mice) but affords protection following LPS challenge. AB - Twitcher mice have an autosomal recessive mutation in the gene for the lysosomal enzyme galactosylceramidase, which is the same gene that is affected in human globoid cell leukodystrophy (Krabbe's disease). The failure to digest galactosylceramide and psychosine leads to initial pathological changes in oligodendrocytes. Secondary pathological changes that include infiltrating macrophages and other inflammatory responses have been postulated to promote the disease course. TNFalpha levels are elevated in twitcher mice compared to control animals, and studies on another demyelinating disease, experimental allergic encephalomyelitis, indicate that TNF promotes pathogenesis via TNF-receptor 1 (TNF-R1). In the present study, twitcher/TNF-R1 deficient mice were generated, and the clinical and pathological course was compared between these mice and regular twitcher mice. There was no statistical evidence for any differences between these two groups of mice for all clinical (life span, weight loss, onset day of twitching) and pathological (demyelination, astrocyte gliosis, macrophage infiltration) measures that were examined. If mice were administered an intraperitoneal injection of LPS, then twitcher/TNF-R1 deficient mice had a longer [corrected] life span and a decreased [corrected] disruption to the blood brain barrier compared to regular twitcher mice. These results showed that TNF-R1 is not sufficiently activated to affect the pathological and/or clinical signs during the natural course of this disease. However, when there is a secondary insult, TNF-R1 activation does lead to a significant acceleration of the development of clinical and pathological signs. PMID- 11024551 TI - Anti-interleukin-3 and anti-nerve growth factor increase neonatal mice survival to reovirus type 3 clone 9 per oral challenge. AB - Reovirus type 3 clone 9 (T3C9) induces lethal encephalitis in neonatal, but not adult mice. Whether host factors that promote the development and/or functioning of nervous and gastrointestinal tissues could modulate the pathogenesis of this enteric virus was examined. The results showed that antibody specific for interleukin-3 or nerve growth factor antiserum, but not anti-interleukin-6 or anti-tumor necrosis factor-alpha/beta increased mice survival to T3C9 and decreased viral titers in nervous tissues early after infection. These data suggest that IL-3 and NGF are involved in the pathogenesis of T3C9 infection in neonatal mice. PMID- 11024550 TI - Modulation of experimental autoimmune encephalomyelitis: effect of altered peptide ligand on chemokine and chemokine receptor expression. AB - Experimental autoimmune encephalomyelitis (EAE) is a T helper 1 (Th1) cell mediated demyelinating disease and the principal animal model for multiple sclerosis. Spinal cords from SJL mice primed with proteolipid protein peptide 139 151 (pPLP) expressed the chemokines RANTES, MCP-1, MIP-2, KC, MIP-1alpha, MIP 1beta, Mig, and fractalkine. We also identified IP-10 in these samples and described a sequence polymorphism in this transcript. Chemokine expression was specific for tissues of the central nervous system. MCP-1, IP-10, and MIP-2 RNA expression significantly correlated with clinical score. Chemokine receptor expression generally correlated with ligand expression. pPLP-primed mice expressed the Th1-associated markers CCR5 and CXCR3 on mononuclear cells. In addition, cells expressing CCR1, CCR2, CCR3, CCR4, CCR8, and CXCR2 were detected. Here we demonstrate that altered peptide ligand (APL)-induced protection from EAE was accompanied by modulation of chemokine and chemokine receptor expression. Spinal cord tissue sections from APL-protected mice showed greatly reduced levels of all chemokines and of CCR1, CCR5, CCR8, CXCR2 and CXCR3. The Th2-associated chemokine receptors CCR3 and CCR4 were found in protected mice, supporting the hypothesis that Th1 but not Th2 cells are down-regulated by APL treatment. This report concludes that chemokines and chemokine receptors can be useful tools to follow modulation of autoimmune disease. PMID- 11024552 TI - Dual effect of glucocorticoids on apoptosis of human autoreactive and foreign antigen-specific T cells. AB - The efficacy of glucocorticoids in the treatment of multiple sclerosis may involve the induction of T cell apoptosis. Here, we report that glucocorticoids have two different effects on the vulnerability of human antigen-specific T cells: (i) steroids induce T cell apoptosis in a CD95-independent, but caspase dependent manner; (ii) steroids protect T cells from CD95-mediated apoptosis which, however, is also caspase-dependent. An increase in BCL-2 expression is observed upon incubation with steroids. Thus, inhibition of CD95-mediated T cell apoptosis may be an undesirable side-effect resulting in survival of activated T cells and the maintenance of pathogenic immune responses might explain the lack of long-term glucocorticoid therapy. PMID- 11024553 TI - Stress induces increases in IL-6 production by leucocytes of patients with the chronic inflammatory disease juvenile rheumatoid arthritis: a putative role for alpha(1)-adrenergic receptors. AB - Juvenile rheumatoid arthritis (JRA) is characterized by chronic inflammation of the joints. In the present study we demonstrate that exposure of JRA patients to a noradrenergic stressor (cold pressor test) results in enhanced LPS-induced IL-6 production by peripheral blood cells of these patients. Healthy, age-matched controls had the same rise in norepinephrine, but do not respond with changes in IL-6 production after exposure to the cold pressor test. Moreover, PBMC of patients with JRA express mRNA encoding alpha(1)-adrenergic receptors (AR), predominantly of the alpha(1d)-AR subtype. In contrast, we could not detect mRNA encoding for alpha(1)-AR in PBMC of healthy controls. The results of this study suggest that expression of alpha(1)-AR mRNA in PBMC during chronic inflammation is associated with altered responses of the immune system to stress. PMID- 11024554 TI - Role of MIP-1beta and RANTES in HIV-1 infection of microglia: inhibition of infection and induction by IFNbeta. AB - Microglia are the major target of HIV-1 infection in the brain. Microglial infection is CD4-dependent, but the role of chemokine receptors CCR5 and CCR3 and their natural ligands in modulating HIV-1 infection in microglia has been questioned. In primary human fetal microglial cultures, we demonstrate that HIV-1 infection of these cells is dependent on CCR5, since an antibody to CCR5 completely blocked productive infection. Anti-CCR3, in contrast, had a smaller inhibitory effect which was not statistically significant. The chemokine ligands for CCR5, RANTES and MIP-1beta, also potently inhibited HIV-1 infection in microglia, but the third ligand MIP-1alpha failed to show inhibition. Interestingly, when microglial cultures were treated with antibodies specific to each of these chemokines, HIV-1 infection was enhanced by anti-RANTES and anti MIP-1beta, but not by anti-MIP-1alpha. These results demonstrate the presence of endogenous chemokines that act as endogenous inhibitors of HIV-1 infection in microglia. Additionally, IFNbeta, a known anti-viral cytokine, also provided potent inhibition of viral infection as well as induction of all three chemokines in microglia. These results suggest the possibility that type I interferon can down-modulate microglial HIV-1 infection in vivo by multiple mechanisms. PMID- 11024555 TI - Myelin basic protein intramolecular spreading without disease progression in a patient with multiple sclerosis. AB - A case with stable multiple sclerosis (MS) and T cell responses which initially focused on peptide 16-38 of myelin basic protein (MBP) allowed us to investigate the dynamics of the MBP-specific T cell repertoire and its relationship with disease progression. Epitope mapping experiments and T cell receptor usage of MBP reactive T cell lines (obtained at four distinct time points over a 7-year period) showed a spreading of the response. Transient expansions and persistence of T cells recognizing different MBP epitopes were also detected. The patient's expanded disability status scale and magnetic resonance imaging lesion load remained stable. Our case shows both persistent self-recognitions and determinant spreading in stable MS. This finding suggests that the relationship between dynamics of self-recognition and disease progression is highly complex. PMID- 11024556 TI - Matrix metalloproteinase-9 (MMP-9) in human cerebrospinal fluid (CSF): elevated levels are primarily related to CSF cell count. AB - Matrix metalloproteinase-9 (MMP-9) was investigated by enzyme-linked immunosorbent assay (ELISA) and zymography in 111 paired CSF and serum samples from patients with various neurological disorders. In 20 patients with blood brain barrier (BBB) impairment but normal CSF cell count, elevated levels of MMP 9 were not observed by ELISA measurement. Another 11 patients characterized in the same way, exhibited only slightly increased MMP-9 levels. In contrast, in 12 patients with intact BBB but elevated CSF cell count, MMP-9 was increased too. It was shown by the more sensitive zymography that MMP-9 increased if CSF cell count exceeded five cells per microl. Spearman rank statistics revealed that MMP-9 concentration in CSF correlated with CSF cell count (r=0.755; P<0.0001), but not with CSF/serum albumin ratio (Q(Alb)) (r=0.212; P=0.057), a measure for BBB impairment. Moreover, the CSF/serum MMP-9 ratio (Q(MMP-9)) did not correlate with Q(Alb)(r=0.192; P=0.100). By use of a Boyden chamber, in which granulocytes migrated through a reconstituted basement membrane, it was demonstrated that the MMP-9 concentration in the lower chamber correlated very significantly with the number of accumulated cells (r(2)=0.7692; P<0.0001). The meaning of the increase of MMP-9 in CSF is critically discussed. PMID- 11024557 TI - Myeloid leukemic cells express and secrete bioactive pituitary-sized 23 kDa prolactin. AB - Prolactin (PRL) is a 23 kDa polypeptide hormone of pituitary origin which is of major importance for reproduction. In addition, PRL has immunomodulatory effects and can be produced in small quantities in nonpituitary tissues. To address possible autocrine or paracrine functions of PRL in leukemia, we characterized immunoreactive PRL from the culture medium of leukemic cells. The myeloid cell line Eol-1 expresses the long extrapituitary type mRNA for PRL and synthesizes immunoreactive PRL with a molecular weight of 23 kDa. The biological activity in Eol-1 culture medium was determined using the Nb2 bioassay. This activity co eluted with recombinant human (rh) PRL on an S-200 Sephacryl gel filtration column and could be blocked by anti-PRL antiserum. Western blot analysis and Nb2 bioassays also suggest that acute myelogenous leukemic blasts secrete bioactive 23 kDa PRL in one out of three tested patients. PMID- 11024558 TI - Postnatal regulation of hypothalamic neuropeptide expression by leptin: implications for energy balance and body weight regulation. AB - Leptin is produced mainly by adipose tissue and has been shown to regulate feeding, energy balance and neuroendocrine function. Regulation of energy homeostasis by leptin is thought to be mediated by hypothalamic neuropeptides, at least in adult rodents. The neonatal period is a critical stage of development during which mammals have to optimize caloric intake to support growth and development, as well as maintain body temperature. It is likely that leptin is involved in the transition from preweaning to adult metabolism. To test this hypothesis, we compared the effect of leptin treatment on body weight and adiposity between neonatal and adult mice. We also determined whether well known hypothalamic neuropeptide targets, e.g. neuropeptide Y (NPY), proopiomelanocortin (POMC), agouti-related peptide (AGRP) and cocaine and amphetamine-regulated transcript (CART) were regulated in a pattern consistent with their presumed roles as mediators of leptin action. Once daily intraperitoneal leptin injection for 7 days did not alter body weight, fat content or expression of hypothalamic neuropeptide mRNAs in 10-day-old mice. In contrast, leptin decreased body weight and adiposity, increased CART and suppressed NPY and AGRP mRNA expression in adult mice. These results are consistent with previous studies showing that the timing of leptin's anorectic action develops after weaning. Furthermore, the association between leptin's ability to influence body weight in adult mice but not in neonates, and the regulation of hypothalamic neuropeptide mRNA expression, is consistent with the view that these peptides mediate leptin's effects on energy balance. PMID- 11024559 TI - SOCS-3 expression in leptin-sensitive neurons of the hypothalamus of fed and fasted rats. AB - Treatment of rodents with exogenous leptin increases SOCS-3 mRNA levels in the arcuate nucleus (ARC) and dorsomedial nucleus (DMN) of the hypothalamus. To determine if SOCS-3 gene activity in the hypothalamus could be influenced by changes in physiological levels of circulating leptin, we performed in situ hybridization (ISH) and immunostaining for SOCS-3 expression in fed vs. fasted (48 h) rats. The ARC and DMN were the only regions of the diencephalon that showed SOCS-3 ISH and the autoradiographic ISH signal for SOCS-3 mRNA was visibly less in the ARC and DMN of fasted rats. The ISH signal for SOCS-3 mRNA was decreased 70% in the ARC and 90% in the DMN (to background levels) when animals were fasted (P<0.01), consistent with decreased immunostaining for SOCS-3 protein observed in the fasted rats. Double fluorescence ISH (FISH) analyses showed colocalization of SOCS-3 mRNA with mRNAs for NPY and POMC in the ARC. These findings are consistent with increased leptin signaling to the NPY and POMC neurons in the ARC by physiological levels of circulating leptin during normal feeding. Therefore, changes in SOCS-3 mRNA levels in the ARC and DMN can be viewed as an indicator of relative physiological leptin signaling to the hypothalamus and also identify cells responding directly to leptin signaling through its cognate receptor. PMID- 11024560 TI - Leptin effects on pulsatile gonadotropin releasing hormone secretion from the adult rat hypothalamus and interaction with cocaine and amphetamine regulated transcript peptide and neuropeptide Y. AB - Leptin may act as a negative feedback signal to the hypothalamic control of appetite through suppression of neuropeptide Y (NPY) secretion and stimulation of cocaine and amphetamine regulated transcript (CART). We aimed at studying the effects of leptin, CART and NPY on the hypothalamic control of the pituitary gonadal system. Pulsatile gonadotropin-releasing hormone (GnRH) secretion was studied in vitro using retrochiasmatic hypothalamic explants from adult rats. In the female, GnRH pulse amplitude was significantly increased by leptin (10(-7) M) and CART (10(-6) M) irrespective of the estrus cycle phase while no such effects were seen in the male. The GnRH interpulse interval was not affected in both sexes. Passive immunoneutralization against CART caused a reduction in GnRH pulse amplitude in the female. A slight but significant increase in GnRH pulse amplitude was caused by NPY (10(-7) M) in the female. However, GnRH pulse amplitude was not affected by a Y5-receptor antagonist (10(-6) M) while the interpulse interval was significantly increased as shown previously in the male. The increase in GnRH pulse amplitude caused by leptin was totally prevented by coincubation with an anti-CART antiserum whereas it was not affected by coincubation with the NPY Y5-receptor antagonist (10(-7) M). In conclusion, leptin and NPY show separate permissive effects on GnRH secretion in the adult rat hypothalamus. In both sexes, NPY is prominently involved in the control of the frequency of pulsatile GnRH secretion through the Y5 receptor subtype. Leptin causes a female-specific facilitatory effect on GnRH pulse amplitude which is mediated by CART and which occurs irrespective of the estrus cycle phase. PMID- 11024561 TI - Regulation of in vivo TSH secretion by leptin. AB - Leptin, the product of the ob gene, is a hormone secreted by adipocytes that regulates food intake and energy expenditure. The hypothalamus-pituitary-thyroid axis is markedly influenced by the metabolic status, being suppressed during food deprivation. The aim of the present study was to assess whether leptin can act as a metabolic signal connecting the adipose tissue with the pituitary-thyroid axis. We studied the effect of leptin administration (10 microg, i.c.v.) on spontaneous TSH secretion and TSH responses to TRH in euthyroid and hypothyroid food-deprived rats. Spontaneous TSH secretion was assessed over 6 h with samples taken every 7 min. Administration of leptin to food-deprived euthyroid rats led to a reversal of the inhibitory effect exerted by fasting on spontaneous TSH secretion. This stimulatory effect of leptin on spontaneous TSH appears to be dependent on the thyroid status since it could not be observed in hypothyroid rats. This data suggests that blunted spontaneous TSH secretion in food-deprived rats is a functional and reversible state, and that the decreased leptin concentrations could be the primary event responsible for the suppression of the hypothalamic pituitary-thyroid-axis in food-deprived rats. PMID- 11024562 TI - Morphological evidence for neural interactions between leptin and orexin in the hypothalamus. AB - Both leptin and orexin have been recently discovered as peptides involved in feeding regulation. The morphological evidence of neural interaction between leptin and orexin, one considered to inhibit food intake and the other to stimulate it in the central nervous system (CNS), was studied by use of double immunostaining method. The leptin receptor-like immunoreactive (LR-LI) neurons in the hypothalamic arcuate nucleus and ventromedial nucleus were innervated by orexin-like immunoreactive (OX-LI) neurons. The distribution of LR-LI neurons in the hypothalamus was very similar to that of OX-LI neurons. These results may suggest that leptin and orexin are intimately correlated with each other and that they reciprocally regulate feeding at the hypothalamic level. PMID- 11024563 TI - Dynamic regulation of leptin entry into brain by the blood-brain barrier. AB - Regulation of the transport of leptin across the blood-brain barrier (BBB) may be crucial for its effects on food ingestion and obesity and may be responsible for 'leptin resistance'. This review summarizes current studies of leptin indicating a dynamic role of the BBB. It includes evidence for its susceptibility to change by physiological stimuli such as starvation, refeeding, and time of day. Although the short form of the leptin receptor is involved in leptin transport, it appears that other mechanisms of entry also exist. Regardless, the BBB is intimately involved with the regulation of the actions of leptin. PMID- 11024565 TI - Leptin: a possible link between food intake, energy expenditure, and reproductive function. AB - Several regulatory substances participate in the regulation of both food intake/energy metabolism and reproduction in mammals. Most of these neuropeptides originate and act in the central nervous system, mainly at specific hypothalamic areas. Leptin represents a signal integrating all these functions, but originating from the periphery (adipose tissue) and carrying information mainly to central structures. Observations in rodent models of leptin deficiency have suggested that leptin participates in the control of reproduction, in conjunction with that of food intake and energy expenditure. Indeed, leptin administration resulted in the restoration of normal body weight, food intake, and fertility in the ob mouse, lacking circulating leptin. Specific targets of leptin in the hypothalamus are neurons expressing neuropeptide Y, proopiomelanocortin and gonadotropin-releasing hormone, but the presence of leptin receptors in peripheral reproductive structures suggests that leptin might also act at these sites. Human obesity is often associated with reproductive disturbances. The situation in humans is more complex than in the animal models of leptin deficit and the presence of leptin resistance in these subjects is suggested. In conclusion, leptin fits many requirements for a molecule linking the regulation of energy balance and the control of reproduction. PMID- 11024564 TI - Response of leptin mRNA to 24-h food deprivation and refeeding is influenced by age in rats. AB - To obtain an insight into the influence of aging on leptin gene expression, the responses of leptin mRNA in retroperitoneal and epididymal adipose tissues and plasma leptin concentrations to 24-h food deprivation and refeeding were examined in 2-, 10- and 24-month-old normal rats. The basal level of leptin gene expression in retroperitoneal adipose tissue was significantly higher in 10- and 24-month-old rats than that in 2-month-old rats, while the level in epididymal adipose tissue was highest in 10-month-old rats for all three age groups. The basal concentrations of plasma leptin was significantly higher in 10- and 24 month-old rats than those in 2-month-old rats. The 24-h food deprivation was followed by a significant reduction in leptin mRNA expression in both retorperitoneal and epididymal adipose tissues for all three age groups. The leptin gene expression was restored to control levels 24 h following refeeding in the 2- and 10-month-old rats, but failed to be restored in the 24-month-old rats. In addition, the time course of recovery for leptin mRNA expression by refeeding to the control levels differed between the retroperitoneal and the epididymal adipose tissue in 2- and 10-month-old rats. The concentrations of plasma leptin 24 h following refeeding were compatible with the leptin mRNA levels in adipose tissues in three age groups. These results suggest that the expression of the leptin gene in response to food-deprivation and refeeding is influenced by an animal's age and that this expression is different for different regions of white adipose tissue. PMID- 11024566 TI - Single intracerebroventricular bolus injection of a recombinant adenovirus expressing leptin results in reduction of food intake and body weight in both lean and obese Zucker fa/fa rats. AB - Leptin acts as a satiety factor within the central nervous system by binding to its receptor located in the hypothalamus. A missense mutation of the leptin receptor induces hyperphagia and obesity in the obese Zucker fa/fa rat. Since the CNS is an important target of leptin action, we hypothesized that leptin gene transfer into the lateral cerebral ventricle could efficiently lead to inhibition of food intake and reduction of body weight in obese fa/fa rats as well as in lean animals. A single intracerebroventricular injection of an adenoviral vector containing a cDNA encoding leptin resulted in the expression of leptin in the ependymal cells lining the ventricle and the secretion of leptin into the cerebrospinal fluid (CSF). During the first week after injection, when high concentrations of leptin were produced in the CSF, the reducing effects of leptin on food intake and body weight were comparable in lean and in obese fa/fa rats. The subsequent decline in CSF leptin levels, that was similar in lean and obese fa/fa rats, resulted in the faster resumption of food intake and body weight gain in obese than in lean animals, confirming a reduced sensitivity to leptin in the obese group. The results of this study show that leptin gene delivery into the cerebral ventricles allows for the production of elevated leptin concentrations in CSF, and they support the hypothesis that the impaired sensitivity to leptin may be overcome in obese fa/fa rats. PMID- 11024567 TI - Resistance to the anorexic and thermogenic effects of centrally administrated leptin in obese aged rats. AB - The aim of the present study was to determine whether the anorexic and thermogenic effects of leptin were attenuated in overweight aged rats following intracerebroventricular (i.c.v.) injection of murine leptin. Male F344/BN rats of two ages (6 months: young (n=20) and 24 months: old (n=18)) were divided into three groups (control, pair-fed and leptin) and were treated with either vehicle (artificial cerebrospinal fluid) or leptin (15.6 microgram/day) for 3 days. There was an age-related increase in basal food intake (20+/-2%), serum leptin levels (363+/-106%) and leptin (OB) mRNA (72+/-16%) in perirenal white adipose tissue (PWAT). In contrast, basal expression of hypothalamic NPY mRNA and brown adipose tissue (BAT) uncoupling protein 1 (UCP1) mRNA was reduced significantly (-35+/-4% and -51+/-5%, respectively) with age. I.c.v. leptin treatment had a significantly greater effect in reducing food intake (-42+/-5% vs. -23+/-4%), serum leptin levels (-55+/-7% vs. 10+/-2%) and PWAT OB mRNA (-46+/-2% vs. 10+/-5%) in young than in old rats. Similarly, central leptin treatment also had a greater effect in suppressing hypothalamic NPY mRNA expression in young (-23+/-4%) than in old ( 8+/-4%) rats compared with their age-matched pair-fed treated rats. The stimulatory effect of i.c.v. leptin treatment on BAT UCP1 mRNA expression was also significantly greater in young rats (45+/-8%) than in old rats (10+/-6%) compared with age-matched pair-fed rats. Our previous report indicated that these overweight aged rats were resistant to peripheral administered leptin. The present data extend those findings and demonstrate that the impaired anorexic and metabolic effects of leptin are centrally mediated. This leptin resistance may be due to either the elevated obesity and serum leptin with age or due to age itself or both. The development of leptin resistance with age may contribute to the hyperphagia, hyperleptinemia and impaired energy balance with age. PMID- 11024568 TI - Leptin is a potent stimulator of bone growth in ob/ob mice. AB - Leptin, the product of the obese gene, is a circulating hormone secreted primarily from adipocytes. The lack of leptin in ob/ob mice, who are homozygous for the obese gene, results in hyperglycemia, hyperinsulinemia, hyperphagia, obesity, infertility, decreased brain size and decreased stature. To this end, we investigated the role of leptin as a hormonal regulator of bone growth. Leptin administration led to a significant increase in femoral length, total body bone area, bone mineral content and bone density in ob/ob mice as compared to vehicle treated controls. The increase in total body bone mass was a result of an increase in both trabecular and cortical bone mass. These results suggest that the decreased stature of the ob/ob mouse is due to a developmental defect that is readily reversible upon leptin administration. Our demonstration that the signalling or long form (Ob-Rb) of the leptin receptor is present in both primary adult osteoblasts and chondrocytes suggests that the growth promoting effects of leptin could be direct. In summary, these results indicate a novel role for leptin in skeletal bone growth and development. PMID- 11024569 TI - Synergistic interaction between CCK and leptin to regulate food intake. AB - Leptin administered (either intracerebroventricularly, icv, or intraperitoneally, ip) acts in synergy with CCK to suppress food intake and body weight in lean mice or rats. The potentiating effect induced by the co-injection of ip CCK and leptin to inhibit food consumption in mice is mediated by the CCK-A receptor and capsaicin sensitive afferents. In vitro, studies in rats showed that a subset of gastric vagal afferent fibers responded to leptin injected directly into the gastric artery only after a prior intra-arterial CCK injection. Moreover, the tonic activity of gastric-related neurons in the nucleus tractus solitarius (NTS) increased when leptin was delivered into the gastric chamber of an in vitro stomach-brainstem preparation. CCK co-injected with leptin potentiated Fos expression selectively in the area postrema, NTS and paraventricular nucleus of the hypothalamus (PVN), which points to the PVN as part of the afferent and efferent limbs of the circuitry involved in the synergistic interaction between leptin and CCK. The dampening of CCK or leptin inhibitory action on ingestive behavior when either factor is not present or their receptors are non functional supports the notion that such leptin-CCK interaction may have a physiological relevance. These observations provide a mean through which leptin and CCK integrate short- and mid-term meal-related input signals into long-term control of energy balance. PMID- 11024570 TI - Leptin physiology: a second look. AB - It is widely believed that the primary physiologic role of leptin is to prevent obesity by regulating food intake and thermogenesis through actions on hypothalamic centers. Here we sugest that the first premise, the anti-obesity role, is untenable, and present evidence for an alternative physiologic role, namely antisteatotic activity in which fatty acid overaccumulation in nonadipose tissues is prevented by leptin-mediated regulation of beta-oxidation. The second premise, namely that leptin acts exclusively on the hypothalamus, is confirmed in normal lean animals with plasma leptin concentrations below 5 ng/ml; their correlation with cerebrospinal fluid levels supports the classical concept of leptin-mediated hypothalamic regulation of food intake. However, when chronic hyperleptinemia exceeds 15 ng/ml, as in obesity, a further rise in plasma leptin does not raise cerebrospinal leptin levels or reduce food intake. Nevertheless, the peripheral antisteatotic action of leptin in acquired obesity continues, suggesting that at chronically hyperleptinemic levels the hormone acts primarily on peripheral tissues and that its hypothalamic action has reached a plateau. PMID- 11024571 TI - Long-term differential modulation of genes encoding orexigenic and anorexigenic peptides by leptin delivered by rAAV vector in ob/ob mice. Relationship with body weight change. AB - We investigated the long-term effects of physiological levels of leptin produced by gene therapy on body weight (BW) and expression of genes that encode orexigenic and anorexigenic peptides in the hypothalamus. Recombinant adeno associated viral vector (rAAV), a non-pathogenic and non-immunogenic vector, encoding leptin (betaOb) was generated and administered iv to ob/ob mice lacking endogenous leptin. Whereas the lowest dose of rAAV-betaOb (6x10(9) particles) was ineffective, the middle dose (6x10(10) particles) curbed BW gain without affecting food consumption for 75 days of observation. A ten-fold higher dose (6x10(11) particles) resulted in increased blood leptin levels and suppressed both BW gain and food consumption throughout the duration of the experiment. rAAV betaOb doses that either curbed BW without affecting food consumption or evoked BW loss and reduced food intake, decreased the expression of genes encoding the orexigenic peptides, neuropeptide Y and agouti-related peptide in the ARC, and the two doses were equally effective. Concomitantly, the expression of genes encoding the anorexigenic peptide, alpha-melanocyte stimulating hormone and cocaine-and-amphetamine regulatory transcript, was augmented with the latter gene displaying a dose-dependant response. These results document the efficacy of delivering biologically active leptin for extended periods by an iv injection of rAAV-betaOb and show that physiological leptin concentrations simultaneously exert a tonic inhibitory effect on orexigenic and a stimulatory effect on anorexigenic signaling in the hypothalamus. This intricate dynamic interplay induced by leptin regulates BW with or without an effect on food intake in leptin deficient ob/ob mice. Further, these results suggest that gene therapy is an effective mode of delivery to the hypothalamus of those therapeutic proteins that cross the blood-brain barrier to ameliorate neuroendocrine disorders. PMID- 11024572 TI - Regulation of leptin secretion: effects of aging on daily patterns of serum leptin and food consumption. AB - We have investigated the effects of age on the daily rise in serum leptin levels during the dark-phase of the light-dark cycle. The results show that in young 7 week-old rats, serum leptin levels increase significantly at 2300 h from the levels at 1500 h in association with increased food consumption. However, in middle-aged rats 25 weeks old, the dark-phase increase in serum leptin is absent despite retention of the daily dark-phase increase in food consumption. When compared to our earlier published results, these finding show that the loss of dark-phase rise in serum leptin occurred despite the daily increase in adipocyte leptin gene expression. These results are in accord with the view that the daily pattern in serum leptin is unlikely to be a contributor to the daily patterning of food consumption. PMID- 11024573 TI - Chronic leptin administration promotes lipid utilization until fat mass is greatly reduced and preserves lean mass of normal female rats. AB - Leptin is a hormone synthesized and secreted from adipose tissue. To study the physiologic effects of chronic leptin treatment, normal adult female Sprague Dawley rats were injected subcutaneously for 35 days. Twice daily injections (250 microgram/day, b.i.d.) resulted in a significant (P<0.05) decrease in food intake that was maintained for 10 days before gradually returning to control level by day 21. Leptin decreased body weight by a maximum of 12% of the initial body weight on day 22 and remained reduced for the duration of the treatment. After 35 days of treatment, visible peritoneal adipose tissue was not detected. Body composition analysis showed that chronic injection of leptin resulted in a dramatic decrease in fat content (28+/-2 to 4+/-2 g, P<0.05; mean+/-SEM) while the lean content remained unchanged. Rats pair-fed to the leptin-treated group but treated with vehicle had the same body composition (23+/-3 g fat mass) as that measured for the ad libitum fed controls. Using indirect calorimetry we observed that leptin decreased respiratory quotient and thus increased fat oxidation. Leptin also prevented energy expenditure reduction typically associated with food restriction. Leptin treatment for 35 days decreased plasma triglyceride (0.75+/-0.07 to 0.30+/-0.03 mM, P<0.05), free fatty acid (0.56+/ 0.06 to 0.32+/-0.04 mM) and insulin (3.2+/-0.5 to 1. 4+/-0.4 ng/ml, P<0.05) concentrations despite the fact that food intake was normalized by day 35. Withdrawal of leptin triggered hyperphagia indicating that leptin biology remained throughout the duration of the chronic treatment. These data suggest that leptin reduces fat mass by initially decreasing appetite and by maintaining enhanced fat utilization even when food intake has returned to that of vehicle treated control. PMID- 11024574 TI - The effects on auditory neurocognitive evoked responses and contingent negative variation activity of frontal cortex lesions or ablations in man: three new case studies. AB - Our previous research in patients with extensive surgical ablations of the prefrontal cortex contradict the hypothesis of some authors that the generators of several auditory event-related potentials (ERPs) (N100; P200; N200; P300; SW), recordable in humans with depth/scalp electrodes and MEG over the prefrontal dorsolateral cortical areas, are essentially located in medial prefrontal and anterior cingulate-limbic cortices. Using a standard CNV paradigm, 21 EEG electrodes and topographic mapping analysis, the post-warning (S1) auditory N100a b c, P200, P300 (binaural clicks) and CNV activity were recorded in three additional patients after extensive dorsolateral and/or medial prefrontal cortex ablations, verified through CT/MRI examinations. No true post-S1/CNV components were recordable over the ablated frontal areas, only sporadic volume-conducted ERPs probably generated in the temporo-parietal lobes or posterior cingulate gyrus. For one of these patients, after excision of a vast right frontal epileptogenic cortical region (including extensive dorsolateral areas, but sparing the fronto-medial cortex and anterior/middle cingulate gyrus), no post S1/CNV components were recordable over the ablated regions. These latest observations again indicate that independent neuronal generators of several post S1 auditory and CNV components are located in the dorsolateral supramodal premotor/prefrontal cortical areas which are directly, ipsilaterally connected to the uni/multimodal temporo-parieto-occipital sensory and associative regions through the long, two-way, fairly superficial, superior arcuate-longitudinal and deeper superior and inferior occipito-frontal bundles. Clear and almost constant differences in the latency of some post-S1 N100 subcomponents (especially the time-lapses between onset and the highest amplitude of the N100 a and c) over various posterior, central and anterior cortical areas sequentially involved, roughly measured in 10 normal subjects along the scalp and with MRI cerebral imaging, may probably be accounted for by the transcortical homohemispheric conduction time, which varies in our scalp recordings from 1 cm/0.74-1.28 ms, mean approximately 1 cm/1.02 ms ( approximately 9.8 ms). PMID- 11024575 TI - On electromyographic (EMG) gradients and movement-related brain activity: significance for motor control, cognitive functions, and certain psychopathologies. AB - Electromyographic (EMG) voltage that rises continuously during motor performance or mental activity and falls precipitately at the end is known as an EMG gradient. Our review is based on 55 studies of EMG gradients, which were published during the period 1937-1994. The extremely wide diversity of situations yielding EMG gradients suggests the possibility that these gradients may be universal accompaniments of organized goal-directed behavioral sequences, overt and covert. Motor tasks and cognitive tasks (without any requirements for motor output) were found to have this in common: they both produced EMG gradients. EMG gradients were not observed during simple, repetitive exercises. On the efferent side, we propose a dual model for the production of EMG gradients, which is based on empirical findings. This model is discussed in relation to current views on central and peripheral neural control of muscle contractions, and on the electrical properties of extrafusal and intrafusal muscle fibers, with particular reference to surface electromyography. The complex relations between EMG gradient steepness and mental effort seemed well represented by the dual model, which was also useful in the interpretation of certain EMG gradients that were found in patients with anxiety disorders, tension-type headaches, and auditory hallucinations, respectively. On the afferent side, drawing on data from human and animal studies, we consider the evidence for movement-related brain activity generated by proprioceptive input, in relation to different types of feedback to the central nervous system during tasks that produce EMG gradients. On electromyographic (EMG) gradients and movement-related brain activity: significance for motor control, cognitive functions, and certain psychopathologies. PMID- 11024576 TI - Current status of diabetes in Taiwan. PMID- 11024577 TI - Diabetes mellitus: perspective from the Asia-Pacific region. AB - This article aims to provide an overview of the current situation in the Western Pacific Region as a prelude to the other articles in this supplememt. It emphasises that, while there is much diversity within this huge Region, there is a rising prevalence of diabetes throughout the Region from which few, if any, countries and areas are spared. This is strongly associated with the lifestyle changes which follow industralisation, mechanisation and massive urbanisation; for which evolutionary processes have left us metabolically ill-equipped. The current epidemic of diabetes is principally due to rises in Type 2 diabetes, although Type 1 diabetes prevalence rates are also rising. The Region already contains more than 30 million people with diabetes and the number is estimated to double by 2025. Diabetes prevalence rates already exceed 8% in 12 countries and areas within the Region. In the more impoverished parts of the Region the majority have little access to care and in all parts of the Region a substantial proportion remain undiagnosed. The increasing emergence of Type 2 diabetes in young adults and children gives particular cause for concern. There is an urgent need for targetted action at all levels within the Region. PMID- 11024578 TI - The epidemiology and management of diabetes mellitus in Indonesia. AB - Predications indicate a potentially explosive increase in the prevalence of diabetes worldwide, especially in developing countries such as Indonesia. Studies of people living in rural areas of East Java and Bali show a prevalence rate of 1.5% in 1982 to 5.7% in 1995 among the urban population. Ujung Pandnag also experienced an increase, and recent studies in Manado found a dramatically high rate of 6.1% in urban areas. Preliminary results indicate varying prevalence between those living in urban and rural areas. Currently, Indonesia has an estimated 1.2-2.3% prevalence among people over 15 years. Geographically variation appears to be an influential factor, due to differences in ethnics, race, culture and lifestyle. Studies of diabetic families show a significantly high prevalence and, clinically speaking, the mode of treatment indicates the type of diabetes. Those who respond well to OHA among young diabetics (<40) are assumed to have the MODY variation of the disease. The level of obesity among the general population has increased, due partly to increased calorie intake and is a significant factor in the increased rate of diabetes. It is also more common among the elderly, as our results will show. The new types of the disease are clinically more difficult to assess than the classical types 1 and 2, as they require relatively costly genetic and immunological studies. The rate of LADA type diabetes was found to be relatively high (>20% for ICA and IAA and 2.3% for GAOA). A concensus on diabetes management has now been formulated in Indonesia and these guidelines are now used by all Indonesian health care professionals. PMID- 11024579 TI - Genetic and environmental interaction in Japanese type 2 diabetics. AB - In Japanese type 2 diabetes, which occupies more than 95%, it is an increasingly important problem as a life-style related disease. The total diabetic population is estimated as approximately 7 million with a prevalence of approximately 6%. Along with genetically low postprandial insulin secretion, they are found to be less tolerable to being overweight to develop insulin resistance. The body weight change in the prediabetic era consisting of 508 male patients treated on their diet alone was reviewed and it was found that a few kilograms of weight gain could be a cause of insulin resistance. Moreover, inactive aldehyde dehydrogenese 2 (ALDH2), which is common in Japanese, is found to be a factor in the development of hyperglycemia. In 163 diabetics, HbAlc of the inactive ALDH2 group was 8.1+/-1.3, while that of active ALDH2 was 7.5+/-0.9% (P<0.05) in a light, social drinking group. However, Japanese type 2 diabetes is also changing. In recent years, the data from a 75-g oral glucose tolerance test of 2121 clients showed that insulinogenic index of clients with impaired glucose tolerance was similar to that of a normal glucose tolerance group and that the area under the insulin curve (AUC) was high in younger diabetics. From a life-style modification perspective, the importance of body weight control by diet and exercise as well as refraining from excessive drinking should be emphasized. PMID- 11024580 TI - Where is Malaysia in the midst of the Asian epidemic of diabetes mellitus? AB - Population studies all over the world have clearly showed that the prevalence of Type 2 diabetes mellitus (DM) is escalating at phenomenal scale and very likely we are heading towards epidemic proportions. In 1985, the estimated population of diabetic individuals in the world was 30 million but by 1995 this figure soared to 135 million. Based on current trends, epidemiologists predict that the population of diabetic individuals will swell up to a staggering 300 million by the year 2025. Almost half of that will be in the Asia Oceania region alone. Dr Hilary King of WHO pointed out that there will be a projected rise of about 42% in developed countries whereas the developing countries will see an escalation to the magnitude of 170% (H. King, R.E. Aubert, W.H. Herman, Global burden of diabetes, 1995-2025: prevalence, numerical estimates and projections, Diabetes Care 21 (1998) 1414-1431; WHO Health Report 1997, WHO Switzerland). There will be a 3-fold rise of the disease in Asia and much of these will be seen in China (40 million) and India (55 million) by virtue of the massive population of these countries. Nevertheless, the other rapidly developing Asian nations like Singapore, Malaysia, Thailand and those making up Indochina will experience the surge. At the same time the prevalence and incidence of diabetes complications will also increase. Based on recent WHO prediction (WHO Newsletter, The global burden of diabetes 1995-2025. World Diabetes 3 (1997) 5-6), it is estimated that by the year 2000 the following figures will be seen:Diabetes complications are major causes of premature death all over the world and most of these are avoidable. DCCT and UKPDS are landmark studies showing strong evidence that major complications can be drastically reduced by maintaining to near normoglycaemic control. PMID- 11024581 TI - Epidemiology of diabetes mellitus in Western pacific region: focus on Philippines. AB - The aim of Diabcare-Asia project was to collect data on diabetes control, management and complication status among patients in 12 Asian countries. Information was extracted from medical records, interviews and laboratory assessment. The majority (96%) of patients were diagnosed with type 2 diabetes mellitus, with mean age (+/-SD) of 59.3+/-12.5 years and mean diabetes duration of 9.4+/-7.0 years. Mean body mass index (BMI) was 24.7+/-4.9 kg/m(2) and the majority (60%) had BMI < or =25 kg/m(2). The majority (70%) of patients were treated with oral antidiabetic drugs (OADs), 15% with insulin, 10% with insulin and OAD combination therapy and 5% with diet control. Among OADs-treated patients, most (44%) received two or more medication. Majority of patients (> or =79%) had satisfactory metabolic control of triglycerides (<2.2 mmol/l), total cholesterol (<6.5 mmol/l) and HDL cholesterol (>0.9 mmol/l). Glucose self monitoring (either urine or blood) was only practiced by 50% of patients. Glycaemic control (HbA1c) was unsatisfactory as majority of patients had HbA1c>7.4% (73%) and 50% had fasting blood glucose (FBG)>7.8 mmol/l. Cataract (26%), neuropathy (42%) and cerebral stroke (6%) were the most frequently reported complications. Clearly, the level of glycaemic control in majority of patients is below satisfaction. Effective education must be emphasised in the management of diabetes. PMID- 11024582 TI - The changing demography of diabetes mellitus in Singapore. AB - Diabetes mellitus has been on the rise in Singapore, while Singaporeans are becoming more affluent, our lifestyles are more sedentary and our population is ageing rapidly. The prevalence of diabetes mellitus rose from 2% in 1975 to 4.7% in 1984, 8.6% in 1992 and 9.0% of adults 18-69 years old in 1998. Malay and Indian women and Indian men were at higher risk, with 14.3, 14.9 and 16.7% prevalence rates, respectively. A further 15% of the adult population have impaired glucose tolerance (IGT). Diabetes was a factor in 39.7% of strokes and in 9.3% of all deaths in Singapore, and is the sixth most common cause of death. In the Diabcare Singapore 1998 Study, 91% of participants were diagnosed with Type 2 diabetes, with mean BMI of 25.1+/-4.4 kg/m(2). The incidence of Type 1 diabetes in childhood is 2.46 per 100000 children 0-12 years of age, while Type 2 diabetes in childhood is an emerging problem. The prevalence of obesity (BMI >30 kg/m(2)) among persons aged 18-69 years rose to 6% in 1998, up from 5.1% in 1992. The prevalence of obesity was highest among the Malays (16.2%) followed by the Indians (12.2%) and the Chinese (3.8%). About 12% of schoolchildren are obese. Increased efforts must be made to change lifestyle and eating patterns in our society, reduce childhood obesity and encourage adults to make lifelong sports and exercise part of the Singaporean way of life. Singapore has one of the world's fastest ageing populations, and even now, 32.4% of Singaporeans 60-69 years of age have diabetes. We should consider screening for diabetes in obese schoolchildren and seek to improve quality of care for people with diabetes, including enlisting the aid of community organisations to improve access to diabetes education, monitoring, support and complications screening services. PMID- 11024583 TI - Genetic epidemiology of type 1 diabetes mellitus in Taiwan. AB - Some environmental and genetic factors play important roles in etiopathogenesis of type 1 or insulin-dependent diabetes mellitus (IDDM). HLA genes, the IDDM1 locus located the human chromosome 6, were found to be associated with insulin dependent diabetes mellitus. However, the incidence of IDDM varied greatly among various populations. To evaluate the pathogenetic factors contributing to the development of IDDM in Taiwan, HLA typing was performed in a group of IDDM unrelated individuals and IDDM pedigrees along with the normal controls from the northern Taiwan. DNA genotypes of class II HLA were done by polymerase-chain reaction based oligotyping techniques. We confirmed that class II HLA genes were significantly associated with IDDM in Taiwan. To study detailed molecular structure of class II HLA molecules and disease association, we examined several amino acid residues on DQalpha and DQbeta chains and the molecular mechanisms to explain the heterozygotic effect of the DR3/DR4 and DR3/DR9 in the Chinese population. Linkage analysis in our pedigrees confirmed the association between HLA and IDDM in population association studies. Among the several class II alleles, a closer segregation of HLA-DQB1*0401 to the affected persons might suggest that HLA-DQB1*0401 itself or an allele closely linked to the DQB1 locus was the IDDM-predisposing allele in Taiwanese. For IDDM2 (INS) region, association with IDDM was not found due to that more than 90% of the population carried class I alleles. In our collection of IDDM, we found few cases (2.4%) carried mitochondrial DNA mutation. Our studies in Taiwanese confirm a multigenetic nature for IDDM. PMID- 11024584 TI - Epidemiologic study of type 2 diabetes in Taiwan. AB - Diabetes mellitus (DM) in adults is a global health problem, although its prevalence varies widely between different populations and the rate has generally increased worldwide. In Taiwan, the mortality rate from DM has almost doubled over the past 10 years. The prevalence of DM in Taiwan was established between 1985 and 1996 and the rates were between 4.9 and 9.2%. The prevalence of impaired glucose tolerance (IGT) was 15.5% (men 15% and women 15.9%). The prevalence of DM and IGT increased significantly with age for both genders. The significant factors associated with newly diagnosed DM were age, BMI, family history of DM, systolic blood pressure (hypertension), physical activity and serum triglyceride levels. The prevalence of large vessel disease (LVD) in DM and non-diabetic subjects were 20.0 and 12.9%, respectively. Among diabetics, 15.8% had ischemic heart disease (IHD), 1.7% leg vessel disease (leg VD), and 2.5% stroke. In non diabetics, the prevalence of the aforementioned macroangiopathies were 11.5, 0.2 and 1.2%, respectively. The diabetics had a significantly higher prevalence of macrovascular disease than non-diabetic subjects. The most significantly associated with the LVD was serum cholesterol levels. Serum cholesterol and HbA1(c) were significantly associated with the development of IHD. Cigarette smoking and female gender were significantly associated with the leg VD. The prevalence of diabetic retinopathy (DR) was 35.0%. (background DR 30%, preproliferative DR 2.8% and proliferative DR 2.2%, respectively.) The prevalence of DR for previously and newly diagnosed diabetics were 45.2 and 28.3% (men 42.8 vs. 33.3% and women 47.5 vs. 24.8%), respectively. From multiple logistic regression analysis, duration of DM was the most important risk factor related to DR. Diabetic subjects treated with insulin had a higher risk of developing retinopathy than those treated with dietary control. The prevalence of nephropathy and neuropathy were 12.9 and 23.5%, respectively. For those patients with and those without nephropathy and neuropathy, the duration of DM, percentage of insulin treatment, percentage of hypertension, and fasting plasma glucose were significantly different. Diabetic duration, hypertension, insulin treatment and glycemic control consistently correlated with nephropathy and neuropathy. In conclusion, the prevalence of DM in Taiwan was between 4.9 and 9.2%, and the prevalence of IGT was 15.5%. The possible risk factors of newly diagnosed diabetes were age, family history of DM, BMI, SBP (hypertension), physical activity and triglyceride levels. Diabetes in Chinese subjects share many characteristics similar to other Asian populations. The burden imposed by the chronic complications of diabetes is massive. In Taiwan, the mortality rates from DM have increased greatly over the past 10 years. Reduction of the modificable risk factors such as BMI, hypertenion and dyslipidemia, and increase of physical activity and good glycemic control through public health efforts may help to reduce the risk of DM and its chronic complications. PMID- 11024585 TI - The incidence of type 2 diabetes mellitus in Taiwan. AB - Currently, diabetes mellitus is the fifth leading cause of death in Taiwan. The trends of diabetes mortality is increasing steadily. Epidemiologic studies also showed increasing prevalence of diabetes mellitus over the past few decades. The incidence of diabetes mellitus in Taiwan has only been studied in recent 10 years. The areas that have been included as study areas for diabetes incidence are Kin-Chen (Kinmen), Chu-Dung, Pu-Tzu, Pu-Li and Pu-Tai. The reported incidence rates ranged from 1.0 to 4.0% per year for people with varying degrees of baseline plasma glucose levels not reaching the diagnosis of diabetes mellitus according to the criteria of the World Health Organization. Age, baseline glucose level, and obesity are important predictors for the development of diabetes mellitus. In the Pu-Tai study, which was aimed at following a group of people who had been living in the hyperendemic villages of blackfoot disease and had been exposed to arsenic from drinking artesian well water, the incidence of diabetes mellitus was calculated to be 27.4 per 1000 person years. The incidence of diabetes mellitus in these arseniasis-hyperendemic villages correlated with age, body mass index and cumulative arsenic exposure. PMID- 11024586 TI - Diabetes in New Zealand. AB - While the ethnic make up of the New Zealand population is predominantly European, the Polynesian population, consisting of indigenous New Zealand Maori and more recent immigrants from the other Pacific Islands is increasing rapidly. The prevalence of diabetes in these Polynesians is high. There is also an increasing prevalence of obesity, and obesity is a greater problem amongst Polynesian people. The number of elderly people in the population is increasing. All of these demographic changes are increasing the incidence and prevalence of Type 2 diabetes. The incidence of Type 1 diabetes is also rising, although the reasons for this are unknown. Diabetic nephropathy is the most common cause of end stage renal failure in New Zealand. Polynesian people with diabetes, and in particular Maori, have a very high rate of diabetic nephropathy and develop renal failure at a more rapid rate than European patients with nephropathy relating to Type 1 diabetes. The propensity for Maori patients with Type 2 diabetes to develop renal failure may relate to a younger age at the onset of diabetes, a genetic susceptibility to nephropathy, and socio-economic or cultural factors leading to less adequate medical care. PMID- 11024588 TI - Diabetes--a common, growing, serious, costly, and potentially preventable public health problem. AB - An estimated 135 million people worldwide had diagnosed diabetes in 1995, and this number is expected to rise to at least 300 million by 2025. The number of people with diabetes will increase by 42% (from 51 to 72 million) in industrialized countries between 1995 and 2025 and by 170% (from 84 to 228 million) in industrializing countries. Several potentially modifiable risk factors are related to diabetes, including insulin resistance, obesity, physical inactivity and dietary factors. Diabetes may be preventable in high-risk groups, but results of ongoing clinical trials are pending. Several efficacious and economically acceptable treatment strategies are currently available (control of glycemia, blood pressure, lipids; early detection and treatment of retinopathy, nephropathy, foot-disease; use of aspirin and ACE inhibitors) to reduce the burden of diabetes complications. Diabetes is a major public health problem and is emerging as a pandemic. While prevention of diabetes may become possible in the future, there is considerable potential now to better utilize existing treatments to reduce diabetes complications. Many countries could benefit from research aimed at better understanding the reasons why existing treatments are under-used and how this can be changed. PMID- 11024587 TI - Type 2 diabetes and the metabolic syndrome in Japanese Americans. AB - Japanese Americans have experienced a higher prevalence of type 2 diabetes than in Japan. Research conducted in Seattle suggests that lifestyle factors associated with 'westernization' play a role in bringing out this susceptibility to diabetes. These lifestyle factors include consumption of a diet higher in saturated fat and reduced physical activity. A consequence of this is the development of central (visceral) adiposity, insulin resistance, and other features associated with this insulin resistance metabolic syndrome, such as dyslipidemia (high triglycerides, low HDL-cholesterol, and small and dense LDL particles), hypertension, and coronary heart disease. We have postulated that the superimposition of insulin resistance upon a genetic background of reduced beta cell reserve results in hyperglycemia and diabetes among Japanese Americans. This article reviews evidence that support this view. PMID- 11024601 TI - Eye burns: an emergency and continuing problem. AB - Adequate treatment of eye burns is an essential task of rescue teams. Clinical and occupational medicine studies have shown that efficient emergency treatment can prevent severe eye damage, but therapy is frequently delayed or inadequate. When initial therapy has been delayed or missed, several treatment strategies, including surgery, are available that may improve the outcome of an injury with poor visual prognosis. Discussed in this review are common accident mechanisms, causative agents and biophysical/pathogenetic aspects of eye burns, together with emergency and long-term treatment strategies including surgical procedures, and factors influencing outcome. PMID- 11024602 TI - Sulphuric acid burned women in Bangladesh: a social and medical problem. AB - In August-September 1998 an Italian medical team, composed of a plastic surgeon, an anaesthesiologist, an infectious diseases specialist and a psychologist, assisted by a local non-government organization (Narripokkho), studied a cohort of Bangladeshi women injured by sulphuric acid. The goals of the mission were: (1) recognition of the magnitude of the phenomenon of acid being intentionally thrown at women; (2) evaluation of the type and severity of burns; (3) preparation of a tentative schedule of surgical treatment of the lesions; (4) organization of local facilities; (5) training of doctors and nurses. PMID- 11024603 TI - Intravenous lidocaine infusion in the treatment of experimental human skin burns digital colour image analysis of erythema development. AB - Previous studies have shown that local anaesthetics possess a wide range of effects on the pathophysiology of burns, including inhibition of burn oedema and inhibition of progressive burn ischemia. The present randomised double-blind cross-over study in six volunteers investigated the effects of intravenous lidocaine infusion on partial thickness skin burns. A thermoprobe was used to induce a standardised thermal injury (1 cm(2)) on the flexor side of one forearm and was repeated on the opposite side 1 week later. Subjects received either an intravenous bolus dose of lidocaine (1 mg kg(-1)) immediately after the thermal trauma followed by continuous intravenous infusion of lidocaine (40 microg kg(-1) min(-1)) during 4 h or equal volumes of isotonic saline. Macrophotographs of the experimental skin area were taken preburn and 1, 2, 3, 4, and 12 h postburn and evaluated by computerised image colour analysis using normalised rgb (n-rgb) and Hue-Saturation-Intensity (HSI) colour systems as a quantitative measure of pathophysiological events. Maximum erythema occurred 2-3 h postburn. Differences between lidocaine- and placebo-treated burns were not significant during the first 4 h postburn. At 12 h postburn, the lidocaine-treated burn demonstrated a significantly faster restitution of residual erythema compared to control sites. The present study shows that intravenous lidocaine significantly inhibits the long-term inflammation-induced tissue responses to thermal trauma. PMID- 11024605 TI - Human beta defensin is absent in burn blister fluid. AB - Defensins are a family of cationic antimicrobial peptides that participate in innate host defence. Human beta defensin-2 (HBD-2) is produced by human keratinocytes, and has a potent bactericidal activity against a wide spectrum of microorganisms. We have recently shown that expression of HBD-2 is present in normal skin and lost in the full-thickness burn wound. Defensins have been found in the blister fluid of chronic wounds. Our study was designed to examine blister fluid from partial-thickness burns for defensin content. Fluid from five patients was collected from partial-thickness burn blisters, and then analysed by sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with a monoclonal antibody and rabbit polyclonal antibody to HBD-2. The assay was validated against a Western blot assay for HBD-2 in samples of bronchoalveolar lavage fluid from patients with inflammatory lung disease. No HBD-2 was detectable in any of the burn blister fluids analysed. HBD-2 is lost in the full-thickness burn wound, and we have now demonstrated its absence in burn blister fluid. This finding represents evidence of a host defence defect within the burn wound and suggests a possible therapeutic role for antimicrobial peptides in the management of burn wounds. PMID- 11024604 TI - Quantification of functional results after facial burns by the faciometer. AB - In the present study the faciometer(R) is introduced in order to quantify the ranges of mimic movements observed after surgical treatment of facial burns. This instrument which consists of calipers and an electronic display was introduced in 1994 in order to measure the extent of facial palsy during reconstructive procedures. The study group consisted of 23 patients, who had been operated on for facial burns. The distances between standardised stable and moving points in the face were determined after mimic movements such as lifting of the eyebrows, maximum showing of the teeth and pursing of the lips. These distances were expressed as a percentage of the distance at rest. For comparison the scars were classified according to the Vancouver Scar Scale. In all patients the functional results after burn trauma in the face and, in some cases, asymmetries at rest could be objectified. Depending upon the severity of scarring, the distance between tragus and mouth was shortened between 0 and 19% after maximal showing of the teeth. In general the mouth region showed more functional deficits than the forehead. Comparing different manners of treatment, it could be objectively demonstrated that the results after deep burns requiring skin grafts were worse than those observed after more superficial lesions and other methods of coverage. The application of keratinocytes to close the burn showed highly variable results. PMID- 11024606 TI - Can blood taken from intraosseous cannulations be used for blood analysis? AB - Intraosseous infusion is increasingly finding application for venous access to infuse drugs and fluids in paediatric resuscitation including burns. The aim of this study was to determine the feasibility of using blood samples taken from the bone marrow cavity for routine blood analysis. In paediatric resuscitation where alternative venous access is not available this could prove valuable. Peripheral venous blood samples were compared with blood obtained from the bone marrow cavity in patients undergoing bone marrow aspiration for the investigation of possible medical pathology. Haemoglobin, haematocrit, sodium, urea, creatinine, and calcium levels values were sufficiently similar to be clinically useful. The potassium level was elevated in most bone marrow samples and the difference from peripheral venous samples variable. Great caution should be exercised in their interpretation. Glucose levels should also be interpreted with care. In conclusion, blood samples obtained intraosseously may give a useful guide to peripheral blood levels of some haematological and biochemical variables. This is potentially useful where a peripheral venous sample cannot be obtained. The values must be interpreted with care. Whether these findings can be extended to paediatric patients who are not haemodynamically stable has not been assessed. PMID- 11024607 TI - Bacteriology in burn patients undergoing mechanical ventilation. AB - In this paper the authors introduce a retrospective study of the incidence of infectious processes in the Burns Unit of the Cruces Hospital (Bilbao), in those patients treated between 1995 and 1998, and who needed, for different reasons, mechanical ventilatory support. The most common microorganisms found in wound cultures, plugged telescoping catheter and blood cultures and analyse variations throughout the stay in the Burns Unit are described. PMID- 11024608 TI - Nosocomial infections in an Iranian burn care center. AB - Burn patients are obviously at high risk for nosocomial infections due to the immunocompromizing effects of burn injury. Pseudomonas aeruginosa is an important life-threatening nosocomial pathogen in burn units. The aim of this study was to determine nosocomial infections in the Tohid Burn Center in Tehran, Iran. Materials of this study were samples of burn wounds and blood from 582 patients who required hospitalization during March 1996 and September 1998. Burn wound samples were taken on admission day, 3 and 7 days after admission. Frequency of culture positive on admission day, 3 and 7 days after admission were 15, 66, and 88%, respectively. Frequency of P. aeruginosa and Staphylococcus aureus on admission day were 35 and 34%, on the third day after admission 73 and 15%, and at the end of the first week of admission 87 and 9%, respectively. Frequency of blood culture positive was 36% (19/53) of which 89% were P. aeruginosa. Overall mortality rate was 18.5% (108/582). Of these patients, frequency of positive wound culture was 92% (99/108). In conclusion, our results show that P. aeruginosa is the leading cause of nosocomial infections in our burn center. It is also necessary to introduce urgent measures for restriction of the spread of P. aeruginosa infections in our burn center. PMID- 11024609 TI - An easy method for preparation of postage stamp autografts. AB - A modified technique of postage stamp autografting is studied. By using quick cutting plates, chessboard tray, skiing technique and petrolatum gauze, skin islands can be uniformly located and correctly oriented on the gauze. Moving the skin squares on the stainless steel tray is relatively easy, and the expansion ratio can be predicted by means of the chessboard diagram. The skin island grafts adhere to the petrolatum gauze when the gauze is applied to the tray. Then, as in the modified Meek technique, the "autografting carrier" can be used to cover the burn wound. In comparison with the modified Meek technique, this method does not require expensive equipment. The quick cutting plates can take over the function of the modified Meek-Wall dermatome. The stainless steel trays and the petrolatum gauze are easily available. The advantages of this method include: (1) the expansion ratio can reach up to 1:9, (2) the preparation procedures are not time consuming, (3) regular distribution of these skin islands shortens the healing time, and (4) the cost of this method is relative low. The results of this technique are comparable to that of the modified Meek technique. This preliminary study reveals that this technique is an easy and economic alternative for the treatment of extensive burn patients. PMID- 11024610 TI - The significance of peripheral skin temperature measurement during the acute phase of burn injury: an illustrative case report. AB - An increase in the gradient between core and peripheral skin temperature is a common finding during the early post burn period which is often interpreted as an indication of inadequate circulating blood volume. In this case study we monitored the pattern of rectal and skin surface (toe) temperature and skin blood flow (by laser Doppler imaging) during the first 16 h after a 34% bath scald in a child (female) of 36 months. In the absence of invasive monitoring of cardiac output, clinical assessment of the adequacy of fluid resuscitation was made from changes in hourly urine volumes and regular (4-h) monitoring of haematocrit. Peripheral vasoconstriction, leading to an increase in the temperature gradient between the inside and outside of the body, was not related to either a fall in urine volume or to haemoconcentration. We suggest that during the first 16 h after burn injury, widening of the core-to-peripheral temperature gradient cannot be used reliably as a clinical indicator of inadequate circulating blood volume. PMID- 11024611 TI - Lime burns in a professional football goalkeeper - an unusual hazard. AB - Burns due to lime, although well reported as a result of occupational exposure, are uncommon in the sports arena. A case of such a chemical burn, which required surgery in a football goalkeeper is presented. There are safe alternatives to line marking on football pitches the use of which will prevent such injuries and avoid litigation. PMID- 11024612 TI - Iatrogenic burn caused by an alcohol lamp. AB - Iatrogenic injury is an intrinsic risk of all medical procedures. Various kinds of iatrogenic complications have been extensively reported and recognized. Two patients were referred to the Prince of Wales Hospital for iatrogenic flame burn caused by the breakage of an alcohol lamp during ENT examination. They were 8 and 9 years old and sustained 12 and 17% intermediate to deep dermal burns respectively. There was no eye or inhalation injury. They were treated initially at the referring hospital. Despite having the best possible treatment, the parents were hostile to and demanding of the medical attendants and the patients were uncooperative throughout the whole treatment period. Rehabilitation of the two children was jeopardized. The situation was very difficult when they were first seen at the Prince of Wales Hospital. The burn surgeon at the Prince of Wales Hospital, as a third party, managed to act as a bridge between the referring hospital and the patients and their families. Several meetings were held and the best interests of the two patients was addressed. The compensation issue was also settled without formal legal prosecution. PMID- 11024613 TI - Chemical burns due to blood pressure cuff sterilized with ethylene oxide. AB - This paper reports two unusual instances of ethylene oxide burns that were caused by the blood pressure cuff sterilized with ethylene oxide. It is suggested that the mechanical compression caused by the blood pressure cuff facilitated penetration of ethylene oxide residues into tissues and contributed to the degree of tissue damage. PMID- 11025134 TI - Shosaiko-to and other Kampo (Japanese herbal) medicines: a review of their immunomodulatory activities. AB - The use of alternative medicine, including consumption of herbal products and dietary supplements, has been increasing substantially both in the United States and in Western Europe. One area that is garnering increased attention is the use of Oriental Medicine including Kampo, or Japanese herbal medicine. Herein, we review representative examples of research available on the most common use of Kampo medicinals, namely to improve the immune response. We also provide an extensive background on the history of Kampo. There are more than 210 different Kampo formulae used in Japan and most uses of Kampo are to modulate the immune response, i.e. to improve immunity. We have extracted data on seven common Kampo medicinals, and the data are reviewed with respect to in vitro and in vivo activities for both humans and experimental animals; the ingredients as well as the problems with classification of these materials are presented. Research suggests that Kampo herbals are biologically active and may have therapeutic potential. While it is believed that Kampo medicines have few side effects, there is a paucity of data on their toxicity as well as a relative lack of knowledge of the bioactive constituents and potential drug interactions of these agents. PMID- 11025135 TI - The wound healing properties of Channa striatus-cetrimide cream-wound contraction and glycosaminoglycan measurement. AB - Haruan has been proved to influence the different phases of wound healing process. The current research focuses on the effects of haruan on the different constituents of extracellular matrix of healing wounds in normal and diabetic rats. Anaesthetized normal and streptozotocin induced diabetic rats were provided with excision wounds at the back and then animals were divided into four groups as: group 1, wounds treated with cetrimide+haruan cream; group 2, wounds treated with haruan cream; group 3, wounds treated with cetrimide (commercial) cream; and group 4, wounds untreated and served as control. Animals were sacrificed after 3, 6, 9 and 12 days. These wounds were used to determine the hexosamine, protein, uronic acid and glycosaminoglycan contents and the wound contraction. The results suggested a marked increase (P<0.05) in the uronic acid, hexosamine and dermatan sulfate contents on day 3 of group 1 when compared with groups 2-4. Wound contraction of group 1 was also markedly enhanced of group 1 (P<0.01) when compared with groups 2- 4. On the basis of these results, we conclude that haruan enhances the synthesis of different glycosaminoglycans in healing wounds, which are the first component of extracellular matrix to be synthesized during the wound healing process. The enhanced levels of glycosaminoglycans may help in the formation of a resistant scar and enhanced wound contraction represents the positive influence of haruan on the fibroplastic phase of wound healing. PMID- 11025137 TI - Anti-diarrheal effect of water extract of Evodiae fructus in mice. AB - Our previous study showed that Evodiae fructus (the dried, unripe fruit of Evodia rutaecarpa) has an inhibitory effect on the intestinal transit (anti-transit effect) in mice. In the present study, a water extract of Evodiae fructus was used to examine its effect on castor oil-induced diarrhea and to compare with its anti-transit effect in mice. The results indicated that Evodiae fructus had both anti-transit and anti-diarrheal effects with comparable ID(50) (the dose for 50% inhibition) values of 54+/-7 and 76+/-17 mg/kg. The time-courses of Evodiae fructus pretreatment for both anti-transit and anti-diarrheal effects were very similar. Because no significant influences of both nitric oxide (NO) precursor L arginine (600 mg/kg, i.p.) and NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (25 mg/kg, i.p.) pretreatment, the NO system was not involved in both the anti-transit and anti-diarrheal effects of Evodiae fructus. Like Evodiae fructus, a muscarinic acetylcholine receptor antagonist atropine inhibited castor oil induced increase in fecal weight and loss of body weight. However, the potencies or time-courses of atropine pretreatment for both anti-transit and anti-diarrheal effects were different. Furthermore, the anti-diarrheal effect of atropine was independent of its anti-transit effect at the lower dose (0.5 mg/kg, i.p.). Therefore, the action of Evodiae fructus appeared to be something different from atropine, suggesting that an action other than the anti-muscarinic action, as previously proposed for Evodiae fructus, may be involved. PMID- 11025136 TI - Inhibition of excitotoxic neuronal death by methanol extract of Acori graminei rhizoma in cultured rat cortical neurons. AB - Acori graminei rhizoma (AGR) are reported to exhibit a number of pharmacological actions in the central nervous system. The effects of the methanol extract of AGR on excitotoxic neuronal death were evaluated in the present study using cultured rat cortical neurons. Based on the phase-contrast microscopic examinations of cultures and lactate dehydrogenase activities measured in the culture media, the glutamate-induced excitotoxicity was significantly inhibited by the extract. The inhibitory action of the extract was more potent and selective for the N-methyl-D aspartate (NMDA) receptor-mediated toxicity. The AGR extract competed with [3H]MDL 105,519 for the specific binding to the glycine site of the NMDA receptor with the IC(50) value of 164.7 microg/ml. Modulation of the NMDA receptor activity by the extract was determined using [3H]MK-801 binding studies. The reduction of the binding in the presence of the extract indicated the receptor inactivation by AGR. These results demonstrated that the methanol extract of AGR exhibited protective action against excitotoxic neuronal death, and that the neuroprotective action was primarily due to the blockade of NMDA receptor function by the interaction with the glycine binding site of the receptor. PMID- 11025138 TI - Pharmacological activity of Abies pindrow. AB - The widely known tree Abies pindrow (Talisapatra) (family: Pinaceae), famous for its diverse clinical uses in Ayurvedic medicines, was investigated to rationalise some of the ancient claims. The petroleum ether (PE), benzene (BE), chloroform (CE), acetone (AE) and ethanol (EE) extracts of A. pindrow leaf were found to have mast cell stabilizing action in rats. The EE, AE and BE extracts offered bronchoprotection against histamine challenge in guinea-pigs. The BE, CE and PE extracts had protective role in aspirin-induced ulcer in rats. The results suggest that while terpenoids, flavonoids, glycosides and steroids are involved in mast cell protection, terpenoids and flavonoids are brochoprotective against histamine-induced bronchospasm. The ulcer protective action of PE, BE and CE fractions of A. pindrow may be attributable to steroids contents only because though all the extracts tested positive for glycosides, the extracts EE and AE did not have any ulcer protective role. PMID- 11025139 TI - The toxic effects of Coleus barbatus B. on the different periods of pregnancy in rats. AB - Extracts of Coleus barbatus B. have been used in folk medicine to interrupt pregnancy. In order to evaluate if this plant interferes with embryo implantation or with the normal development of the concepts, pregnant Wistar rats were treated with increasing doses (220, 440 and 880 mg/kg per day) of a hydroalcoholic extract of C. barbatus. The rats received the extract by gavage from days 0 to 5 of pregnancy (preimplantation period) or 6 to 15 (organogenic period). Control groups received distilled water during the same periods. The animals were killed at term for the evaluation of maternal and fetal parameters. The results showed that the treatment with 880 mg/kg per day of the extract of C. barbatus before embryo implantation caused delayed fetal development and an anti-implantation effect, which justifies the popular use of this extract with abortive purposes. After embryo implantation delayed development associated with maternal toxicity was observed in the fetuses of the group which received 880 mg/kg per day. PMID- 11025140 TI - On the anti-inflammatory and anti-phospholipase A(2) activity of extracts from lanostane-rich species. AB - We have studied extracts from three species rich in lanostane triterpenes for their activity against different in vivo models of inflammation induced by TPA, EPP and PLA(2). The inhibitory effect against PLA(2) in vitro was also studied. When the Poria cocos extract was tested against PLA(2)-induced mouse paw edema, it was active by the oral and parenteral routes. Its effect was greater in both magnitude and duration than that of Pistacia terebinthus and Ganoderma lucidum extracts. P. terebinthus was effective against chronic and acute inflammation, and according to a preliminary chromatographic analysis, its seems to be a good source of lanostane anti-inflammatory agents. G. lucidum was the least effective of the three species studied and, unlike the other two, failed to inhibit the activity of PLA(2) in vitro. PMID- 11025141 TI - Role of bittergourd fruit juice in stz-induced diabetic state in vivo and in vitro. AB - The aqueous juice of bittergourd fruit (BF) (Momordica charantia L.) of the family Cucurbitaceae has been shown to possess hypoglycemic activity. However, the mechanism of its action is not known. Hence in vitro and in vivo experiments were carried out to study the role of BF juice on the diabetic status. The activity of BF juice was tested on STZ treated RIN cells and isolated islets in vitro. It was found that feeding with BF juice caused reduction in STZ-induced hyperglycemia in mice. It markedly reduced the STZ-induced lipid peroxidation in pancreas of mice, RIN cells and islets. Further it also reduced the STZ-induced apoptosis in RIN cells indicating the mode of protection of BF juice on RIN cells, islets and pancreatic beta-cells. Present study thus confirms hypoglycemic effect of BF juice and provides sufficient documentation to define its role and action for its potential and promising use in treating diabetes. PMID- 11025142 TI - Effect of Ocimum sanctum on noise induced changes in neutrophil functions. AB - The effect of ethanolic extract of Ocimum sanctum was studied on the noise stress induced changes in albino rats. Acute noise stress caused leukopenia, increased corticosterone level and enhanced the neutrophil functions as indicated by an increase in the Candida phagocytosis and Nitro blue tetrazolium (NBT) reduction. Pretreatment with the O. sanctum Linn. (OS) extract brought back the stress altered values to normal levels indicating the stress alleviating effect of O. sanctum. PMID- 11025143 TI - Pharmacological basis for the use of peach leaves in constipation. AB - The aqueous crude extract (PPL.Cr) of peach leaves (Prunus persica) was studied for the possible presence of gut stimulatory constituent(s) to rationalize the folkloric use of the plant in constipation. PPL.Cr at the dose of 1-10 mg/ml caused a moderate degree of spasmogenic effect in isolated guinea-pig ileum. Pretreatment of the tissue with atropine (1 M) completely abolished the contractile effect of the plant extract similar to that of acetylcholine which is suggestive of a cholinergic mechanism. In isolated rabbit jejunum preparations, PPL.Cr produced a week spasmogenic effect followed by relaxation of the spontaneous contractions at higher doses. Bioassay-directed fractionation revealed that the spasmogenic activity was separated in the aqueous fraction, while the spasmolytic activity was concentrated in the ethyl acetate fraction. When tested against K(+)-induced contraction, both PPL.Cr and its ethyl acetate fraction (PPL.EtAc) caused a dose-dependent inhibition, suggesting calcium channel blockade (CCB). The presence of CCB in peach leaves was confirmed when pretreatment of the tissue with PPL.EtAc caused a dose-dependent rightward shift in the Ca(2+) dose-response curves, similar to that produced by verapamil. These data indicate that the plant contains spasmogenic (cholinomimetic) and spasmolytic (calcium antagonist) constituents, which are concentrated in the aqueous and ethyl acetate fractions, respectively. Furthermore, the laxative effect of the plant reported in the traditional system of medicine may be partially due to the cholinergic action, which was dominant over the spasmolytic component. PMID- 11025144 TI - Acute diuretic, natriuretic and hypotensive effects of a continuous perfusion of aqueous extract of Urtica dioica in the rat. AB - This study was performed on anaesthetized male Wistar rats that received a continuous intravenous perfusion during 1.25 h of an aqueous extract of aerial parts of Urtica dioica L. (Urticaceae) at a low dose of 4 mg/kg/h or at a high dose of 24 mg/kg/h, or furosemide (control diuretic) at a dose of 2 mg/kg/h. As compared with a control period in each rat, the arterial blood pressure was reduced proportionally to the dose of the perfusion of the plant extract (15 and 38%, P<0.001, respectively). These effects were accompanied by a correlative increase of diuresis (11 and 84%, P<0. 001, respectively) and natriuresis (28 and 143%, P<0.001, respectively). In the rats perfused by furosemide, the arterial blood pressure was reduced by 28% (P<0.001). The diuresis and natriuresis were also increased proportionally in this case (85 and 155%, P<0.001, respectively). Nevertheless, the hypotensive action of U. dioica was reversible during the recovery periods in about 1 h with the lower dose of the plant extract and furosemide, while the effect of the higher dose was persistent, indicating a possible toxic effect. In conclusion, the results demonstrate an acute hypotensive action of U. dioica that indicates a direct effect on the cardiovascular system. Moreover, diuretic and natriuretic effects were also observed, suggesting an action on the renal function. Finally, the plant extract seems to have a toxic effect at the higher dose. PMID- 11025145 TI - Immunomodulatory and anti-inflammatory activity of Picrorhiza scrophulariiflora. AB - Extracts of the rhizomes of Picrorhiza scrophulariiflora Pennell (Scrophulariaceae) were investigated for their in vitro and in vivo immunomodulatory properties. Diethyl ether extracts showed potent inhibitory activity towards the classical pathway of the complement system, the respiratory burst of activated polymorphonuclear leukocytes, and mitogen-induced proliferation of T-lymphocytes. Furthermore, such extracts showed anti inflammatory activity towards carrageenan-induced paw edema. No effects were observed in experimentally induced arthritis in mice. PMID- 11025146 TI - Effects of Astragali root and Hedysari root on the murine B and T cell differentiation. AB - It has traditionally been known in Asian countries that the Astragali and Hedysari roots have similar beneficial effects. They have been applied in clinical practice for immunological diseases, we investigated their effects on specific antibody production, B cells and T cells in BALB/c mice immunized with ovalbumin (OVA). Both the Astragali and Hedysari roots reduced the production of IgG2a in the primary response to OVA. Hedysari root reduced the production of IgG2a in the secondary response. Both reduced the production of IgM, suppressed IL-6 production in spleen cells. It was presumed that the suppression of IgG2a production was induced by the suppression of IL-6, which is an antibody production inducer. It was found that the Hedysari root increased the B cells and activated them, and the Astragali root also tended to increase them. In addition, Hedysari root decreased the proportions of IFN-gamma-producing cells in splenic CD4(+) T lymphocytes. It was shown that the Astragali and Hedysari roots could act as a growth factor of B cells by increasing the proportion of the total B cells and activated B cells. In conclusion, both the Astragali and Hedysari roots showed the same actions, but the latter was more effective. PMID- 11025147 TI - Hepatoprotective effects of Turkish folk remedies on experimental liver injury. AB - Seven plants which are used in Turkish folk medicine were studied for possible hepatoprotective effects. These plants are Carduus acanthoides and C. nutans (Asteraceae), Cichorium intybus (Asteraceae), Fumaria asepalae and F. vailantii (Fumariaceae), Gentiana olivieri (Gentianaceae) and Plantago lanceolata (Plantaginaceae). Stems, bracts and receptaculum of Cynara scolymus were used as natural reference drugs. Effects of the ethanolic extracts were studied using the carbon tetrachloride-induced hepatotoxicity model in rats. The extracts of F. vailantii and G. olivieri significantly prevented the elevation of plasma and hepatic malondialdehyde formation (evidence of lipid peroxidation) as well as enzyme levels (AST and ALT) in acute liver injury, which might be ascribed to their potent hepatoprotective activity. Liver sections were also studied histopathologically to confirm the biochemical results. PMID- 11025148 TI - Antiplasmodial activity of isoflavones from Andira inermis. AB - The stem bark and seeds of Andira inermis, Fabaceae, are employed as a purgative, vermifuge, and febrifuge. In particular, the powdered bark is claimed to be efficacious in intermittent fever. Bioassay-guided fractionation of lipophilic extracts from the stems and leaves yielded six isoflavones: biochanin A, calycosin, formononetin, genistein, pratensein, and prunetin. Calycosin (3', 7 dihydroxy-4'-methoxyisoflavone) and genistein (4',5, 7-trihydroxyisoflavone) have been shown to possess in vitro activity against the chloroquine-sensitive strain poW and the chloroquine-resistant clone Dd2 of Plasmodium falciparum. PMID- 11025149 TI - Inhibition of tumor necrosis factor-alpha-induced apoptosis by Asparagus cochinchinensis in Hep G2 cells. AB - A human hepatoma cell line, Hep G2 cells, is a reliable system for the study of alcohol-induced hepatotoxicity. In this study, we investigated the effect of an aqueous extract of Asparagus cochinchinensis(MERRIL) (Liliaceae) roots (ACAE) on ethanol (EtOH)-induced cytotoxicity in Hep G2 cells. ACAE (1-100 microg/ml) dose dependently inhibited the EtOH-induced tumor necrosis factor-alpha (TNF-alpha) secretion. ACAE (1-100 microg/ml) also inhibited the EtOH and TNF-alpha-induced cytotoxicity. Furthermore, we found that ACAE inhibited the TNF-alpha-induced apoptosis of Hep G2 cells. These results suggest that ACAE may prevent the EtOH induced cytotoxicity through inhibition of the apoptosis of Hep G2 cells. PMID- 11025150 TI - Antiplasmodial activity of plant extracts used in west African traditional medicine. AB - Five plants originating from Ivory Coast were selected after an ethnobotanical survey, Alchornea cordifolia, Mitragyna inermis, Nauclea diderrichii, Pterocarpus santalinoides, and Terminalia glaucescens. Traditional healers for the treatment of malaria commonly used these plants. Extracts of these plants were tested on three strains of Plasmodium falciparum, FcB1-Colombia and FcM29-Cameroon (chloroquine-resistant strains) and a Nigerian chloroquine-sensitive strain. Extracts were obtained by preparing decoction in water of the powdered plant, the technique used by most of the traditional healers. A radioactive micromethod allowed the evaluation of the in vitro activity of the extracts on P. falciparum. Concentrations inhibiting 50% of the parasite growth (IC(50)) ranged from 2.34 to more than 500 microg/ml according to the plant. For the most active plants (A. cordifolia and T. glaucescens) ethanol and pentane extracts were made and tested. The IC(50) values obtained for these extracts ranged from 0.35 to 43.40 microg/ml. The stage specificity of the ethanol extracts of A. cordifolia and T. glaucescens and pentane extract of T. glaucescens on the parasite erythrocytic cycle were determined. The ethanol extract of T. glaucescens showed its highest activity at the transition from the trophozoite to the schizont stages. Cytotoxicity was estimated on human fibroblasts (HeLa) cells and a cytotoxicity/antiplasmodial index was calculated, it ranged between 5 and 21, and the best antiplasmodial extract (T. glaucescens ethanol extract) had the higher index (>20). PMID- 11025151 TI - In vitro pharmacological activity of the tetrahydroisoquinoline salsolinol present in products from Theobroma cacao L. like cocoa and chocolate. AB - Cocoa and chocolate contain the tetrahydroisoquinoline alkaloid salsolinol up to a concentration of 25 microg/g. Salsolinol is a dopaminergic active compound which binds to the D(2) receptor family, especially to the D(3) receptor with a K(i) of 0.48+/-0.021 micromol/l. It inhibits the formation of cyclic AMP and the release of beta-endorphin and ACTH in a pituitary cell system. Taking the detected concentration and the pharmacological properties into account, salsolinol seems to be one of the main psychoactive compounds present in cocoa and chocolate and might be included in chocolate addiction. PMID- 11025153 TI - Isolation of an antibacterial sesquiterpenoid from Warburgia salutaris. AB - The bark of Warburgia salutaris is used in traditional medicine as an expectorant and smoked for coughs and colds, including a topical application for sores and inflammation. A previous screening of South African medicinal plants showed that this plant had promising antibacterial activity. Subsequently, this endangered tree species was selected for bioassay-guided fractionation in order to identify the active principles. Fractionation of the ethyl acetate extract of the stem bark by chromatographic techniques yielded a sesquiterpenoid which exhibited antimicrobial activity against Gram-positive bacteria. The compound, muzigadial, has previously been reported in two other Warburgia species, this being the first time it has been reported from W. salutaris. Muzigadial had minimum inhibitory concentration values ranging from 12.5 to 100 microg ml(-1). PMID- 11025152 TI - Antimicrobial activity of extracts of eastern North American hardwood trees and relation to traditional medicine. AB - Wood and bark extracts of 14 eastern North American hardwood tree species which were used traditionally as medicine by First Nation's people were screened for antimicrobial activities with eight strains of bacteria and six strains of fungi. Eighty-six percent of the bark extracts were active against methicillin sensitive Staphylococcus aureus; 71% against Bacillus subtilus and 79% against Mycobacterium phlei. The bark extract of Juglans cinerea was active against Pseudomonas aeruginosa 187, Salmonella typhiumurium, and Klebsiella pneumoniae. The wood extracts were less active: 72% were active against S. aureus (methicillin-sensitive), 36% against B. subtilus and 43% against M. phlei. Results from antifungal tests indicated that 36% of the extracts were active against at least one fungal strain and that bark extracts were more active than wood extracts. The bark extract from Juglans cinerea had the broadest spectrum of activities against Candida albicans, Saccharomyces cerevisiae, Cryptococcus neoformans, Trichophyton mentagrophytes, Microsporum gypseum, and Aspergillus fumigatus. In general, the extracts were more active against gram positive bacteria than gram negative bacteria and against filamentous fungi than yeast like fungi. The study also demonstrated a correlation between frequency of traditional medicinal use by the First Nations people and antimicrobial activity of extracts indicating that the traditional knowledge encompasses an understanding of aspects of chemical ecology. PMID- 11025154 TI - Indigenous knowledge and uses of medicinal plants by local communities of the Kali Gandaki Watershed Area, Nepal. AB - A field survey was conducted in the villages of Ramdi, Malunga, Balam, Beltari, Mirmi, Burgha and Ridi in the Kali Gandaki watershed, Nepal; 48 medicinal plants belonging to 31 families were reported, each with local names, traditional uses, methods of preparation and route of administration. Traditional medicine remains an integral part of the health system in these areas. Local people have remarkable knowledge of species identity and their uses as crude drugs. PMID- 11025155 TI - Anticonvulsant effect of water extract of Scutellariae radix in mice. AB - Since a previous study indicated that the water extract of Scutellariae radix (SR) had high affinity for the benzodiazepine (BDZ) binding site of GABA(A) receptors, the present study examined whether SR water extract has an anticonvulsant effect in vivo and an enhancing effect on gamma-amino-n-butyric acid (GABA)-stimulated uptake of 36Cl(-) in cortex preparation in vitro in mice. The results showed that SR water extract had little effect on pentylenetetrazol (PTZ, 85 mg/kg, s.c.)-induced clonic seizures but significantly inhibited maximal electroshock-induced tonic seizures with an ED(50) of 3.6 g/kg. The BDZ agonist chlordiazepoxide (10 mg/kg, i.p.) had anticonvulsant activity on both types of seizures. In 36Cl(-) uptake assay, SR water extract (1-500 microg/ml) had no significant effect on 25 microM GABA-stimulated 36Cl(-) uptake, whereas chlordiazepoxide (10 microM) increased the 36Cl(-) uptake to 125% of control. Therefore, the present results showed for the first time that SR water extract had anticonvulsant activity against maximal electroshock-induced tonic seizures, and suggested that this anticonvulsant effect might be not via the activation of the BDZ binding site of GABA(A) receptors, but probably via the prevention of seizure spread. PMID- 11025156 TI - Antifungal activity of extracts from medicinal plants used by First Nations Peoples of eastern Canada. AB - From literature describing medicinal usage of plants by First Nations Peoples in eastern Canada, 18 eastern Canadian plants were selected and tested for their antifungal activities. Eight randomly selected tropical plants were also tested for comparative purposes. Four groups of plants were obtained: popular antimicrobial-remedy (n=6), popular non-antimicrobial-remedy (n=6), random temperate (n=6) and random tropical (n=8). Extracts from these plants were tested in disk assays as growth inhibitors of six fungi known to be opportunistic human pathogens (Saccharomyces cerevisiae, Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus, Microsporum gypseum and Trichophyton mentagrophytes). Of the four plant groups tested, extracts from the popular antimicrobial-remedy group were significantly more effective at inhibiting fungal growth based on both overall antifungal activity and number of fungal species inhibited. PMID- 11025157 TI - Inhibition of xanthine oxidase by some Chinese medicinal plants used to treat gout. AB - The enzyme xanthine oxidase catalyses the oxidation of hypoxanthine to xanthine and then to uric acid, which plays a crucial role in gout. A total of 122 traditional Chinese medicinal plants, selected according to the clinical efficacy and prescription frequency for the treatment of gout and other hyperuricemia related disorders, have been evaluated for the enzyme inhibitory activity. Among the 122 methanol extracts derived from these species, 69 were shown to be inhibitory at 100 microg/ml, with 29 having greater than 50% inhibition. As to the equal amount of water extracts, 40 were disclosed to be active at 100 microg/ml, with 13 possessing more than 50% inhibition. At 50 microg/ml, 58 methanol and 24 water extracts exhibited inhibitory activity, with 15 of the former and two of the latter showing greater than 50% inhibition. The most active was the methanol extract of the twig of Cinnamomum cassia (Lauraceae) (IC(50), 18 microg/ml), which was followed immediately by those of the flower of Chrysanthemum indicum (Asteraceae) (IC(50), 22 microg/ml) and the leaves of Lycopus europaeus (Lamiatae) (IC(50), 26 microg/ml). Among the water extracts, the strongest inhibition of the enzyme was observed with that of the rhizome of Polygonum cuspidatum (Polygonaceae) (IC(50), 38 microg/ml). The IC(50) value of allopurinol used as a positive control was 1.06 microg/ml. The study demonstrated that the effects for these medicinal plants used for the gout treatment were based, at least in part, on the xanthine oxidase inhibitory action. PMID- 11025158 TI - Efficiency of traditionally used South African plants against schistosomiasis. AB - Urinary schistosomiasis is treated traditionally by means of herbal remedies. Forty-eight South African plant species were identified as possible antischistosomic plants. Twenty-one of these plant species were collected in order to investigate their antischistosomal properties. Crude extracts of the plant materials were screened against the schistosomula of the species Schistosoma haematobium. Cercariae were obtained from Bulinus africanus snails through an in vitro technique. By subjecting the cercariae to a sheering stress, they were transformed into schistosomula. The schistosomula were placed into a culture medium to which the plant extracts were added. The results obtained indicated that the plant extracts from Berkheya speciosa (Asteraceae), Euclea natalensis (Ebenaceae) and Trichilia emetica (Meliaceae) are lethal to the schistosomula. PMID- 11025159 TI - Phyllanthus amarus extract administration increases the life span of rats with hepatocellular carcinoma. AB - The effect of Phyllanthus amarus extract administration after induction of hepatocellular carcinoma (HCC) by N-nitrosodiethylamine (NDEA) was studied in Wistar rats. Administration of an aqueous extract of P. amarus was found to significantly increase the survival of hepatocellular carcinoma harboring animals. All the untreated rats died of tumour burden by 33.7+/-1.6 weeks. Administration of P. amarus extract (150 mg/kg b.w.) after tumour development increased the survival of animals to an average of 52. 2+/-2.3 weeks. Serum gamma glutamyl transpeptidase activity which was elevated to 182+/-23 U/l by NDEA administration was lowered to 112+/-19 U/l by the administration of P. amarus extract. Similarly elevated glutathione S-transferase activity (1534+/-116 nmol/min per mg protein) and glutathione (20.5+/-2.4 nmol/mg protein) levels in the NDEA administered group were found to be lowered to 1112+/-89 nmol/min per mg protein and 14.2+/-2.2 nmol/mg protein respectively. P. amarus administration was found to be ineffective in controlling the liver weight, elevation of tissue gamma-glutamyl transpeptidase, serum alkaline phosphatase and serum glutamate pyruvate transaminase of HCC harboring animals. PMID- 11025160 TI - Ethnobotanical survey in the Palestinian area: a classification of the healing potential of medicinal plants. AB - An ethnobotanical survey was carried out in the West Bank to evaluate the relative efficacy of the plants used to treat skin diseases and prostate cancer. A total number of 102 informants, 30 years and older and either native born or had been living in the West Bank for more than 30 years, were interviewed using a previously prepared questionnaire. Of about 165 plant species mentioned by the informants, 63 (38.1%) were mentioned by three or more informants. On the basis of their primary uses, 21 of these plants were reported to relieve skin disorders, 17 for urinary system disorders, 16 for gastric disorders, nine for cancer and prostate disorders, eight for arthritis, five for respiratory problems, and five for other ailments. Indices on fidelity levels (FLs), relative popularity level (RPL), and rank-order priority (ROP) were calculated. Plants were classified in two groups: 'popular' (RPL=1) or 'unpopular' (RPL<1). The following plant species were classified as popular in this study: Teucrium polium, Matricaria aurea, Urtica pilulifera, Paronychia argentea, Petroselinum sativum, and Salvia fruticosa. The remaining 57 species were classified as 'unpopular'. Fifty-nine plants were claimed to be effective against cancer and prostate disorders, which include Arum dioscorides, U. pilulifera, Allium sativum, Viscum cruciatum, and Allium cepa. PMID- 11025161 TI - Snakebites and ethnobotany in the northwest region of Colombia. Part III: neutralization of the haemorrhagic effect of Bothrops atrox venom. AB - Thirty-one of 75 extracts of plants used by traditional healers for snakebites, had moderate or high neutralizing ability against the haemorrhagic effect of Bothrops atrox venom from Antioquia and Choco, north-western Colombia. After preincubation of several doses of every extract (7.8-4000 microg/mouse) with six minimum haemorrhagic doses (10 microg) of venom, 12 of them demonstrated 100% neutralizing capacity when the mixture was i.d. injected into mice (18-20 g). These were the stem barks of Brownea rosademonte (Caesalpiniaceae) and Tabebuia rosea (Bignoniaceae); the whole plants of Pleopeltis percussa (Polypodiaceae), Trichomanes elegans (Hymenophyllaceae) and Senna dariensis (Caesalpiniaceae); rhizomes of Heliconia curtispatha (Heliconiaceae); leaves and branches of Bixa orellana (Bixaceae), Philodendron tripartitum (Araceae), Struthanthus orbicularis (Loranthaceae) and Gonzalagunia panamensis (Rubiaceae); the ripe fruits of Citrus limon (Rutaceae); leaves, branches and stem of Ficus nymphaeifolia (Moraceae). Extracts of another 19 species showed moderate neutralization (21-72%) at doses up to 4 mg/mouse, e.g. the whole plants of Aristolochia grandiflora (Aristolochiaceae), Columnea kalbreyeriana (Gesneriaceae), Sida acuta (Malvaceae), Selaginella articulata (Selaginellaceae) and Pseudoelephantopus spicatus (Asteraceae); rhizomes of Renealmia alpinia (Zingiberaceae); the stem of Strychnos xinguensis (Loganiaceae); leaves, branches and stems of Hyptis capitata (Lamiaceae), Ipomoea cairica (Convolvulaceae), Neurolaena lobata (Asteraceae), Ocimum micranthum (Lamiaceae), Piper pulchrum (Piperaceae), Siparuna thecaphora (Monimiaceae), Castilla elastica (Moraceae) and Allamanda cathartica (Apocynaceae); the macerated ripe fruits of Capsicum frutescens (Solanaceae); the unripe fruits of Crescentia cujete (Bignoniaceae); leaves and branches of Piper arboreum (Piperaceae) and Passiflora quadrangularis (Passifloraceae). When the extracts were independently administered by oral, i.p. or i.v. route either before or after an i.d. venom injection (10 microg), neutralization of haemorrhage dropped below 25% for all the extracts. Additionally, B. rosademonte and P. percussa extracts were able to inhibit the proteolytic activity of B. atrox venom on casein. PMID- 11025162 TI - Seasonal effect on Brazilian propolis antibacterial activity. AB - The behavior of microorganisms towards the antibiotic action of propolis has been widely investigated. Since reports dealing with seasonal effect on propolis activity are not available, this assay was carried out aiming to observe the in vitro antimicrobial activity of propolis, collected during the four seasons, on bacterial strains isolated from human infections. Dilution of ethanolic extract of propolis (EEP) in agar was the method performed, with serial concentrations ranging from 0.4 to 14.0% (% v/v). The behavior of some bacteria was analysed according to the incubation period in medium plus propolis, and the survival curve was plotted. It was verified that the growth of Gram-positive bacteria is inhibited by low propolis concentrations (0.4%) whereas Gram-negative bacteria were less susceptible to this substance, the minimal inhibitory concentration ranging from 4.5 to 8.0%. There was no significant difference with regards to the seasonal effect on the survival curve of Staphylococcus aureus and Escherichia coli; after incubation with propolis, there was an efficient antimicrobial action, mainly towards Gram-positive bacteria. PMID- 11025163 TI - Effects of chard (Beta vulgaris L. var. Cicla) extract on pancreatic B cells in streptozotocin-diabetic rats: a morphological and biochemical study. AB - Chard (Beta vulgaris L. var. cicla) is used as a hypoglycemic agent by diabetic patients in Turkey. The present study was carried out in order to detect whether this plant, used in folk remedies for decreasing blood glucose levels, affects pancreatic B cells and blood glucose. In the diabetic group, a decrease in the number of B cells of Langerhans islets and in the secretory materials, a swollen granular endoplasmic reticulum cisternae and widened intercellular areas in some of B cells were observed. But, in a diabetic group given chard extract, an increase in the number of B cells of Langerhans islets and in the secretory granules were noted, together with many hypertrophic Golgi apparatus and granules of low densities. The extract while having no effect on blood glucose and body weight in the normal group, reduced the blood glucose value in streptozotocin induced hyperglycemic animals. But, in a diabetic group given chard, the body weight significantly increased in comparison to the diabetic group; maximum reduction in blood glucose levels was observed on the 42nd day. According to the morphological and biochemical results obtained, it is concluded that the extract of this plant when administered by gavage may reduce blood glucose levels by regeneration of the B cells. PMID- 11025164 TI - Psychopharmacological screening of Pfaffia glomerata Spreng. (Amarathanceae) in rodents. AB - The alcoholic extract of Pfaffia glomerata roots (100, 500, 1000 mg/kg, intraperitoneally (i.p.), and 500, 1000, 1500 mg/kg, per os) was studied in several behavioral animal models for the evaluation of central activity: open field, barbiturate sleeping time, pentilenotetrazole (PTZ)-induced convulsions, elevated plus-maze, step-down inhibitory avoidance and forced swimming test. The acute treatment (500 mg/kg, i.p.) interfered with the open-field habituation, decreased sleep latency and increased barbiturate-induced sleeping time, protected partially the animals of PTZ-induced convulsions, decreased the memory retention in step-down inhibitory avoidance, and did not have an important effect in the elevated plus-maze test and forced swimming test. The same extract at 1000 mg/kg per os did not cause any effect in barbiturate sleeping time and pentilenotetrazole-induced convulsions models. Thus, the effect on the memory was deeper evaluated in the step-down inhibitory avoidance task. When administered by intraperitoneal route, the extract showed a dose-dependent effect causing full amnesia at 1000 mg/kg. On the other hand, when it was given by oral route at 500, 1000 and 1500 mg/kg, no influence on the memory retention was observed. These results suggest that the alcoholic extract of P. glomerata roots presents different effects depending on the route of administration: by i.p route, it seems to be a central nervous system depressant agent; by oral route, it seems to be ineffective, at least in the tested doses. PMID- 11025165 TI - A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part IV. Is a new haem polymerisation inhibition test pertinent for the detection of antimalarial natural products? AB - The search for new antimalarial agents in plant crude extracts using traditional screening tests is time-consuming and expensive. New in vitro alternative techniques, based on specific metabolic or enzymatic process, have recently been developed to circumvent testing of antimalarial activity in parasite culture. The haem polymerisation inhibition test (HPIA) was proposed as a possible routine in vitro assay for the detection of antimalarial activity in natural products. A total of 178 plant extracts from the Pharmacopeia of the Bolivian ethnia Tacana, were screened for their ability to inhibit the polymerisation of haematin. Five extracts from Aloysia virgata (Ruiz & Pavon) A.L. Jussieu (Verbenaceae), Bixa orellana L. (Bixaceae), Caesalpinia pluviosa D.C. (Caesalpiniaceae), Mascagnia stannea (Griseb) Nied. (Malpighiaceae) and Trichilia pleenea (Adr. Jussieu) (Meliaceae) demonstrated more than 70% inhibition of haematin polymerisation at 2.5 mg/ml. The extracts were also tested for antimalarial activity in culture against F32 strain (chloroquine-sensitive) and D2 strain (chloroquine-resistant) of Plasmodium falciparum and in vivo against P. berghei. The extract from Caesalpinia pluviosa was the only one that showed activity in HPIA and in the classical test in culture. The accuracy and pertinence of HPIA, applied to natural products is discussed. PMID- 11025166 TI - Antidepressant effects of Banxia Houpu decoction, a traditional Chinese medicinal empirical formula. AB - Banxia Houpu Decoction, having been used for the treatment of depression-related diseases since ancient times, is a traditional Chinese medicinal empirical formula consisting of Pinellia ternata, Poria cocos, Magnolia officinalis, Perilla frutescens and Zingiber officinale. The effects of the total decoction extract and five fractions therefrom, were evaluated in mice by tail suspension and forced swimming tests. The total 90% ethanol extract of the decoction was shown to possess an antidepressant activity that was close to that of Prozac, an antidepressant agent being applied clinically. Furthermore, the active principles were desmonstrated to be mainly in the aqueous (Bx4) and lipophic (Bx5) parts of the decoction extract while the polyphenol fraction (Bx2) exhibited a moderate action. PMID- 11025167 TI - Phytochemical and pharmacological studies on Monechma ciliatum. AB - The leaves of Monechma ciliatum were found to contain alkaloids, glycosides, proteins, tannins and saponins. The hot methanol extract (HME) of the leaves, previously found to have potent oxytocic effect in various species was fractionated in various solvents and the fractions analysed chemically and also assayed on the rat uterus in an attempt to identify the chemical constituents present and in particular the oxytocic principle. Positive reaction with ninhydrin spray suggests that the oxytocic constituent is an amino acidic derivative. PMID- 11025168 TI - Evaluation of some Moroccan medicinal plant extracts for larvicidal activity. AB - The larvicidal properties of 16 extracts of four Moroccan medicinal plants: Calotropis procera (Wild.), Cotula cinerea (L.), Solanum sodomaeum (L.) and Solanum elaeagnifolium (CAV.) were tested against Anopheles labranchiae mosquito larvae. Among the extracts tested, nine exhibited high larvicidal activity with LC(50) (24 h) ranging from 28 to 325 ppm. PMID- 11025169 TI - Antimicrobial activity of some coumarin containing herbal plants growing in Finland. AB - Antimicrobial screening against selected Gram-positive and Gram-negative bacteria, yeasts, mold, as well as plant pathogenic fungi, with emphasis on method optimization was carried out on methanol extracts prepared from seven plants grown in Finland. Sensitivity to the extracts was found to vary considerably among the micro-organisms, the extract from Petroselinum crispum and Ruta graveolens showing the highest toxicity against Rhizoctonia solani. The growth of Heterobasidium annosum was inhibited, whereas that of Phytophtora (cactorum) was promoted by all the extracts. The antibacterial and antifungal activities of six natural coumarin compounds were weak, except for the inhibitory effect against Fusarium culmorum. PMID- 11025170 TI - Preliminary studies on the analgesic activity of latex of Calotropris procera. AB - In this study we have evaluated the analgesic activity of dry latex (DL) of Calotropis procera. A single oral dose of DL ranging from 165 to 830 mg/kg produced a significant dose dependent analgesic effect against acetic acid induced writhings. The effect of DL at a dose of 415 mg/kg was more pronounced as compared to a 100 mg/kg oral dose of aspirin. On the other hand DL (830 mg/kg) produced marginal analgesia in a tail-flick model which was comparable to aspirin. The analgesic effect of DL was delayed by 1 h by naloxone at a dose of 0. 5 mg/kg, i.p., which completely blocked the analgesic effect of morphine (10 mg/kg, i.p.). However, the effect of aspirin was not blocked by naloxone. The 830 mg/kg oral dose of DL did not produce toxic effects in mice and the LD(50) was found to be 3 g/kg. PMID- 11025171 TI - Plant sources for the treatment of jaundice in the tribals of Western Madhya Pradesh of India. AB - A survey of the tribals of Western Madhya Pradesh of India for the treatment of jaundice has yielded 13 plants for this purpose. Part of the plant used, dosage, duration, restriction on intake of food etc. has been reported. PMID- 11025172 TI - Antimicrobial activity of flavonoids in medicinal plants from Tafi del Valle (Tucuman, Argentina). AB - Preliminary studies of flavonoids have been realised in five native species from Tafi del Valle (Tucuman, Argentina) used in popular medicine. Most of compounds detected were flavonoids mono and dihydroxylated in B ring. Screening for antimicrobial activity against Gram positive and Gram negative microorganisms has been realised with Lippia turbinata, Satureja parvifolia, Sambucus peruviana, Verbena officinalis and Chenopodium graveolens. The total extracts of flavonoids of each plant were tested and four species studied showed antimicrobial activity. PMID- 11025173 TI - Inhibitory effects of butanol fraction of the aqueous extract of Forsythia koreana on the nitric oxide production by murine macrophage-like RAW 264.7 cells. AB - The aim of this study was to investigate the effect of butanol fraction of the aqueous extract of Forsythia koreana fruits on the nitric oxide (NO) production and inducible nitric oxide synthesis (iNOS) gene expression in murine macrophage like RAW 264.7 cells. Butanol fraction alone affected neither NO production nor iNOS gene expression in macrophage-like RAW 264.7 cells. However, the butanol fraction inhibited NO production and iNOS gene expression in RAW 264. 7 cells stimulated with interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS). These findings suggest that inhibition of NO production by this butanol fraction in RAW 264.7 cells stimulated with IFN-gamma plus LPS was due to the suppression of iNOS gene expression. PMID- 11025174 TI - 4-Hydroxybenzaldehyde from Gastrodia elata B1. is active in the antioxidation and GABAergic neuromodulation of the rat brain. AB - Ether fraction of G. elata methanol extract significantly inhibited the recovery time and severity induced by pentylenetetrazole (PTZ) treatment. Pretreatment of ether fraction of G. elata methanol extract successfully prevented diminution of brain GABA level in subconvulsive dose of PTZ-treated rats. 4 Hydroxybenzaldehyde, an analogue of p-hydroxybenzyl alcohol, showed an inhibitory effect on the GABA transaminase, and its inhibitory activity was higher than that of valproic acid, a known anticonvulsant. In the brain of PTZ-treated rats, brain lipid peroxidation was significantly increased, while it recovered to the control level after treatment with 4-hydroxybenzaldehyde. It may be concluded that antioxidation and positive modulation of GABAergic neuromodulation of 4 hydroxybenzaldehyde partially contribute to an antiepileptic and anticonvulsive activity of G. elata B1. PMID- 11025175 TI - In vitro activity of Rutaceae species against the trypomastigote form of Trypanosoma cruzi. AB - The activity of crude plant extracts of nine species of Rutaceae against the trypomastigote form of Trypanosoma cruzi was evaluated at 4 mg/ml. Thirty-two crude extracts were tested and eight of them showed significant activity (>80%). The most active extract was obtained from the stems of Pilocarpus spicatus (97.3%). Fractionation of the active crude extracts provided 25 fractions which were tested against the trypomastigote form of T. cruzi at 2 mg/ml. Of these six showed significant activity (>80%). The most active fractions (100%) were obtained from the leaves of Almeidea coerulea (butanol fraction) and Conchocarpus inopinatus (dichloromethane fraction). PMID- 11025176 TI - Evaluation of antiseptic-impregnated central venous catheters for prevention of catheter-related infection in intensive care unit patients. AB - Central venous catheterization represents a significant medical advancement, particularly in the treatment of critical ill. However, there is a high risk of central venous catheters-related infection. A novel antiseptic central venous catheter, made of polyurethane and impregnated with chlorhexidine and silver sulfadiazine, was developed to reduce the risk of catheters-related infection. In this study, we did a randomized clinical study to determine the efficacy by using antiseptic catheters for the prevention of central venous catheters-related infection in the intensive care units. A total of 204 patients with 235 central venous catheters were studied at the surgical intensive care units at National Taiwan University Hospital between November 1998 and June 1999. Participants received either a standard triple-lumen polyurethane catheter or an antiseptic catheter (Arrow International, Reading, Pennsylvania, USA). Both were indistinguishable from each other. Compared to standard polyurethane catheters, antiseptic catheters were less likely to be colonized by microorganisms when they were cultured at the removal (8.0 versus 20.0 colonized catheters per 100 catheters; relative risk 0.34 [95% CI, 0.15 to 0.74]; p<0.01). There was no significant differences between both groups in catheter-related infections (0.9 versus 4.9 infections per 100 catheters; relative risk 0.17 [95% CI, 0.03 to 1.15]; p = 0.07). Gram-positive cocci and fungi were more likely to colonize in the standard polyurethane catheters (p = 0.06 and 0.04, compared to antiseptic catheters respectively). Two of our cases in the control group died directly due to catheter-related candidemia. No adverse reactions such as hypersensitivity or leukopenia were found in the antiseptic catheter group. Our study showed that central venous catheters with antiseptic coating were safe and had less risk of colonization of bacteria and fungi than standard catheters in the critically ill patients. PMID- 11025177 TI - Hepatitis G virus (HGV) infection in voluntary and commercial blood donors in India. AB - The prevalence of hepatitis G virus (HGV) has been determined in commercial as well as voluntary blood donors from India by detecting viral RNA genome using reverse transcription polymerase chain reaction (RT-PCR) assay. 45 professional blood donors from private blood banks and 50 healthy controls who opted for voluntary blood donation were recruited for the study. Both the groups were also screened serologically for HBV and HCV infection. The prevalence of HGV in the general population in India was found to be 4% but significantly a higher frequency (46.6%; p<0.001) of HGV was observed in commercial blood donors. PMID- 11025178 TI - Susceptibility testing of Candida species: comparison of NCCLS microdilution method with Fungitest. AB - Fungitest is a new commercially available and easy-to-perform breakpoint test system using six antifungal agents. We compared this test with a modified standard method described by the National Committee for Clinical Laboratory Standards (NCCLS). One hundred isolates of Candida species were tested with both methods. Based on the same breakpoints, the correlation of qualitative results between the reference method and Fungitest was high. Best results were obtained after incubation of Fungitest for 48 h. Overall agreement was high, an excellent correlation was given with amphotericin B and flucytosine (100% and 99%, respectively), whereas itraconazole showed only 86% concordance. When Fungitest was read after 24 h the agreement was lower ranging from 100% to 75%. Some of the breakpoints used with Fungitest differ from the breakpoints recommended by NCCLS, whereas others have not been elaborated by the NCCLS. The adaptation of Fungitest breakpoints to NCCLS and determination of further breakpoints have to be discussed before Fungitest can be recommended for routine use. PMID- 11025179 TI - Treatment with oxoglaucine can enhance host resistance to Candida albicans infection of mice with adjuvant arthritis. AB - The alkaloid oxoglaucine reduced CD4+ cell clones in adult mice and decreased CD4+, CD8+ and Ig+ levels in newborn mice. It prevented the increase of CD8+ and Ig+ clones induced by Candida albicans (C. albicans) in adult mice. TNF-alpha serum accumulation was inhibited by oxoglaucine in C. albicans infection and adjuvant arthritis. Treatment with oxoglaucine of arthritic mice, followed by inoculation with C. albicans enhanced the host resistance against the pathogen. PMID- 11025180 TI - Sustained gastrointestinal colonization and systemic dissemination by Candida albicans, Candida tropicalis and Candida parapsilosis in adult mice. AB - The ability of nine clinical isolates of Candida species (three C. albicans, three C. tropicalis and three C. parapsilosis) to colonize and invade the gastrointestinal (GI) tract of adult male CD-1 (ICR) mice was determined. The effect of dietary tetracycline plus glucose supplementation on colonization was evaluated. Strains were intragastrically inoculated. Tetracycline and glucose altered substantially aerobic flora, especially streptococci (average fall 1.1 +/ 0.3 log(10) CFU/g, p<0.01 by the Student's t test). At two weeks after oral challenge, sustained and high colonization of GI tract by Candida (mean 5,28 +/- 0.18 log(10) CFU/g, p<0.01) was achieved in all mice receiving glucose tetracycline supplementation, excepting in animals inoculated with one of C. tropicalis isolates. Histologic sections of the stomachs revealed multiple intraepithelial micro-abscesses associated with hyphae in the region of the cardial-atrium fold. Under immunosuppression, systemic spread of C. albicans and C. tropicalis was observed in 62% and 24% of animals receiving dietary supplementation respectively. Dissemination was not noted for C. parapsilosis isolates. We have developed a simple and inexpensive murine model of sustained colonization of GI tract. This model could be useful for analyzing prophylaxis, treatment and diagnosis of systemic Candida infections and for evaluating virulence of strains. PMID- 11025181 TI - Commercial systems for fluconazole susceptibility testing of yeasts: comparison with the broth microdilution method. AB - Fluconazole susceptibility was tested in 100 clinical yeast isolates (65 Candida albicans, 13 C. glabrata, 8 C. tropicalis, 7 C. parapsilosis, 3 Saccharomyces cerevisiae, 1 each of C. krusei, C. lusitaniae, Cryptococcus neoformans, Rhodotorula glutinis) and two control strains (Candida krusei ATCC 6258, C. parapsilosis ATCC 22019) using broth microdilution (reference method), disk diffusion, Etest strips, Sensititre YeastOne, Candifast, Integral System Yeasts. Using M27-A breakpoints, isolates were classified as susceptible (81%), susceptible-dose dependent or Resistant with broth dilution. Rates of concordance with the reference method were good for Sensititre YeastOne, Etest and disc diffusion (81.2%-94.7%) but very low for the Candifast (3.1%) and Integral System (16.6%), which classified most susceptible isolates as resistant. Lack of standardisation (inoculum, medium composition) and non-objective interpretation schemes may be the cause of their poor performance. Sensititre YeastOne, Etest and disc-diffusion are potentially useful for fluconazole antifungal susceptibility testing of yeasts in clinical laboratories. PMID- 11025182 TI - Activity of piperacillin/tazobactam in combination with amikacin, ciprofloxacin, and trovafloxacin against Pseudomonas aeruginosa by time-kill. AB - Pseudomonal infections have a high rate of morbidity and mortality, thus combination therapy is often recommended. We compared the activity of piperacillin/tazobactam in combination with amikacin, ciprofloxacin, or trovafloxacin at different concentrations against P. aeruginosa using time-kill methodology. MICs were determined for 4 clinical isolates of P. aeruginosa. Time kill studies were conducted over 24 h. Each drug was tested alone and in combination using the following concentrations: 2 and 1/4, 1/4 and 2, and 1/4 and 1/4xMIC of piperacillin/tazobactam and amikacin, ciprofloxacin, or trovafloxacin. Combinations were classified as synergistic, indifferent, or antagonistic. Synergy was defined as > or = 2-log(10) decrease in CFU/mL at 24 h with the combination when compared to the most active single agent and the number of surviving organisms for the antimicrobial combination was > or =2-log(10) less than the initial inoculum. The MICs for piperacillin/tazobactam, amikacin, ciprofloxacin, and trovafloxacin, ranged from 4/4-512/4, 0.5-4, 0.125-4, and 0.5 8 microg/mL, respectively. Fifty eight percent of the combinations using concentrations of 1/4xMIC of piperacillin/tazobactam and 2xMIC of amikacin, ciprofloxacin, and trovafloxacin or 2xMIC of piperacillin/tazobactam and 1/4xMIC of amikacin, ciprofloxacin, and trovafloxacin were synergistic. Although no differences existed in synergistic activity between the two combinations, the 1/4 and 2xMIC maintained colony counts below the limit of quantification for 24 h for a significantly greater percentage of isolates than the 2 and 1/4xMIC combinations (75 and 25%, respectively; p = 0.04). Overall, synergy was most frequently (42%) noted with the piperacillin/tazobactam and amikacin combinations followed by 33 and 8% of the piperacillin/tazobactam and trovafloxacin and ciprofloxacin combinations. No combination demonstrated antagonism. Further more extensive studies are necessary to determine clinical significance. PMID- 11025183 TI - Evaluation of antibiotic synergy against Acinetobacter baumannii: a comparison with Etest, time-kill, and checkerboard methods. AB - Acinetobacter baumannii is becoming increasingly resistant to antibiotics, often requiring combination therapy. Numerous methods exist to detect the presence of in vitro synergy with the time-kill and checkerboard tests being widely used. The Epsilometer test (E test) is a new method that is less labor intensive, but has not been evaluated using a wide range of antimicrobials and organisms. We assessed synergy using the time-kill and checkerboard tests and compared the results to the E test method using 10 clinical isolates of A. baumannii. Antimicrobial combinations evaluated consisted of trovafloxacin or tobramycin in combination with cefepime or piperacillin. Synergy was detected with all combinations by either the checkerboard or time-kill method. Synergy was not detected by the Etest method. The agreement between the time-kill test and Etest method was 72% (range 42-97%); for the time-kill and checkerboard tests, agreement was 51% (range 30-67%). The Etest method appears promising although further testing should be performed with additional antimicrobial agents and organisms. PMID- 11025184 TI - Daptomycin susceptibility tests: interpretive criteria, quality control, and effect of calcium on in vitro tests. AB - Daptomycin MICs were determined for 844 Gram-positive bacteria in three concentrations of Ca(++) and compared with the MICs of vancomycin and teicoplanin. Daptomycin was twofold to fourfold more active against most species when tested in 50 microg/ml of Ca(++) than in 25 microg/ml. In 50 microg/ml of Ca(++) daptomycin was more active against methicillin-resistant staphylococci and vancomycin-resistant enterococci than teicoplanin or vancomycin; 100% of these isolates were susceptible to < or =2.0 microg/ml of daptomycin. Different lots of Mueller-Hinton agar were variable in Ca(++) content, and daptomycin disk diffusion zone diameters were affected, i.e., zones were 1 to 15 mm smaller on one lot of agar with only 6 microg/ml of Ca(++) compared to another lot with 28 microg/ml. The previously proposed daptomycin interpretive breakpoints performed satisfactorily when MICs were determined in Mueller-Hinton broth with 50 microg/ml of Ca(++) and when the agar gave appropriate zones with quality control strains. To define those control limits, replicate tests with four quality control strains were performed in ten laboratories using broth microdilution tests (with Ca(++) supplemented broth) and disk diffusion tests on Mueller-Hinton agar without cation adjustments. PMID- 11025186 TI - Effects of underfeeding during the weaning period on growth, metabolism, and hormonal adjustments in the piglet. PMID- 11025185 TI - Ability of laboratories to detect emerging antimicrobial resistance in nosocomial pathogens: a survey of project ICARE laboratories. AB - A proficiency testing project was conducted among 48 microbiology laboratories participating in Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology). All laboratories correctly identified the Staphylococcus aureus challenge strain as oxacillin- resistant and an Enterococcus faecium strain as vancomycin-resistant. Thirty-one (97%) of 32 laboratories correctly reported the Streptococcus pneumoniae strain as erythromycin-resistant. All laboratories testing the Pseudomonas aeruginosa strain against ciprofloxacin or ofloxacin correctly reported the organism as resistant. Of 40 laboratories, 30 (75%) correctly reported resistant MICs or zone sizes for the imipenem- and meropenem resistant Serratia marcescens. For the extended-spectrum beta-lactamase (ESBL) producing strain of Klebsiella pneumoniae, 18 (42%) of 43 laboratories testing ceftazidime correctly reported ceftazidime MICs in the resistant range. These results suggest that current testing generally produces accurate results, although some laboratories have difficulty detecting resistance to carbapenems and extended-spectrum cephalosporins. This highlights the need for monitoring how well susceptibility test systems in clinical laboratories detect emerging resistance. PMID- 11025187 TI - Modulation of growth performance in disease: reactive nitrogen compounds and their impact on cell proteins. AB - During life, all animals encounter situations that challenge their capability for optimal growth. In reacting to immune challenges in the form of disease, homeostatic mechanisms attempt to overcome disharmony of the body's internal environment, or simply put, stress. The overall impact of stress revolves around a dynamic relationship between the level of challenge imparted on physiological systems and the degree of host response that is mounted in the process of detecting and reacting to the stress. In growing animals, the majority of milder stress encounters are manifest in terms of energetic inefficiencies and periods of reduced anabolism. In contrast, severe stress is often characterized by frank catabolism and tissue wasting. In some instances a level of stress (that might be termed a "stress breakpoint") is reached at which time the host response itself contributes to the cascade of negative effectors that further cause illness. These "breakpoint" responses are characterized by more intense acute responses to stress or a much more protracted duration of the response than would be expected given the nature of the stress. Key to understanding how growth in the young animal responds to infectious stresses is the recognition that (a) when immune responses that normally maintain health go awry, the reporters and effectors of the immune system (cytokines and the nitric oxide cascade) can contribute to stress disease processes and (b) reactive nitrogen compounds derived from the nitric oxide, as well as super oxide anion can modify intracellular proteins and block otherwise normal biochemical processes that regulate cell function. A key example of this is the loss of regulation of IGF-I by GH. As animals react more severely to disease stress, IGF-I concentrations in plasma decline progressively. Recent data derived from (LPS) challenges performed on young calves suggest that the prolonged decline in IGF-I is associated with the development of hepatic cytotoxicity localized to regions of protein nitration as identified by immunohistochemistry. Identifying biochemical criteria for disease processes provides needed guidance for the further development of intervention strategies to limit the impact of disease on growth. PMID- 11025188 TI - Reduced growth of calves and its reversal by use of anabolic agents. AB - Disease has profound effects on the immune system, endocrine system, and on the growth process. Since diseases are catabolic to the animal, there is current interest in the possible role of anabolic hormones to counter the effects of disease in general and minimize the effects of a disease process on growth and development. A number of anabolic hormones, such as growth hormone (GH) and estradiol + progesterone (EP), have been studied for their role in enhancing growth and stimulating immune function and are thus candidates for hormonal intervention in disease processes. GH has been shown to be effective in countering some of the deleterious effects of endotoxemia but was ineffective in a parasitic disease model. Studies with EP have shown similar success with both endotoxemia and a parasitic disease model. Moreover, GH and EP do not share a common mechanism of action, suggesting that the effects are not simply due to anabolic actions. While the mechanism of action of GH in endotoxemia has been examined, the effects of EP are via an unknown mechanism, possibly by inhibition of IL-I action or inhibition of nitric oxide overproduction. PMID- 11025189 TI - High body weight gain and reduced bovine mammary growth: physiological basis and implications for milk yield potential. AB - Available evidence concerning the relationship between growth rate, mammary growth and milk yield in heifers leads to these conclusions: 1) Increased growth rate due to high feeding level before puberty onset can lead to reduced pubertal mammary growth and reduced milk yield potential. 2) Increased growth rate due to high feeding level after puberty and during pregnancy have no effect on mammary growth and milk yield. 3) Higher body weight gain due to higher genetic potential for growth is positively related to milk yield. The negative effect of high feeding level before puberty occurs in all breeds, but the level of feeding causing reduced yield varies. Variation in responses between experiments suggests that feeding regimes that support high growth rates without negative effect on yield can be developed. A breakthrough most likely will originate from increased knowledge of the physiological relationship between nutrition and mammary development. Our investigations suggest that blood growth hormone (GH) is important for mammary development, and that the negative effect of high feeding level on mammary development may be due to reduced blood GH. GH, however, does not bind to mammary tissue. Experiments with exogenous GH suggest that GH acts on mammary tissue via IGF-I, but IGF-I is increased by high feeding level - not decreased as GH. This paradoxical relationship cannot be explained by changes in circulating IGF binding proteins. However, the sensitivity of mammary tissue to IGF-I is reduced by high feeding level, probably due to the action of locally produced binding proteins and/or growth factors. PMID- 11025190 TI - Genetically caused retarded growth in animals. AB - Growth process of animals is regulated by a multitude of physiological pathways among which components of the somatotropic axis play a key role. A number of severe, simply inherited growth disturbances have been identified in humans, laboratory and farm animals. These disorders are controlled by defective alleles at major loci referring to hormones or hormone receptors, e.g. growth hormone receptor for the recessive sex-linked dwarfism (dw) in chickens and the recessive autosomal Laron-type dwarfism in man, and growth hormone releasing hormone receptor for the recessive "little" mutation (lit) in mice. Apart from these particular cases, growth rate is a quantitative polygenic trait which has a moderate heritability (close to 0.30) and is influenced by prenatal and postnatal maternal effects. Increase in the average coefficient of inbreeding in a population is also known to result in lower growth rate. Divergent selection experiments have shown that upward or downward selection on growth is effective, sometimes with asymmetrical responses, but patterns of changes in underlying physiological traits appear to differ among experiments. PMID- 11025191 TI - Mechanisms of reduced and compensatory growth. AB - Growth is an integrated process, resulting from the response of cells dependent on the endocrine status and nutrient availability. During feed restriction, the production and secretion of growth hormone (GH) by the pituitary gland are enhanced, but the number of GH receptors decreases. Changes of GH binding proteins induce GH resistance and are followed by reduced insulin-like growth factor-I (IGF-I) secretion. On the other hand, high circulating levels of GH enhance the mobilization of fatty acids, which are used to support energy requirements. Thus, when feed restriction in growing animals is moderate, there is mainly protein but barely fat accretion. By contrast, a severe feed restriction enhances the release of catabolic hormones and stimulates, from muscle cells, the liberation of amino acids, which are used by hepatocytes for gluconeogenesis. During refeeding and compensatory growth, the secretion of insulin is sharply enhanced and plasma GH concentrations remain high. This situation probably allows more nutrients to be used for growth processes. The role of plasma IGF-I during compensatory growth is not clear and must be explained in connection with changes of its binding proteins. Thyroxin and 3,5,3' triiodothyronine seem to have a permissive effect on growth. The simultaneous occurrence of puberty with refeeding can exert a synergistic effect on growth. Initially, compensatory growth is characterized by the deposition of very lean tissue, similar as during feed restriction. This lasts for some weeks. Then, protein synthesis decreases and high feed intake leads to increased fat deposition. PMID- 11025192 TI - Nitric oxide-dependent modification of the sarcoplasmic reticulum Ca-ATPase: localization of cysteine target sites. AB - Skeletal muscle contraction and relaxation is modulated through the reaction of sarcoplasmic reticulum (SR) protein thiols with reactive oxygen and nitrogen species. Here, we have utilized high-performance liquid chromatography electrospray mass spectrometry and a specific thiol-labeling procedure to identify and quantify cysteine residues of the SR Ca-ATPase that are modified by exposure to nitric oxide (NO). NO and/or NO-derived species inactivate the SR Ca ATPase and modify a broad spectrum of cysteine residues with highest reactivities towards Cys364, Cys670, and Cys471. The selectivity of NO and NO-derived species towards the SR Ca-ATPase thiols is different from that of peroxynitrite. The efficiency of NO at thiol modification is significantly higher compared with that of peroxynitrite. Hence, NO has the potential to modulate muscle contraction through chemical reaction with the SR Ca-ATPase in vivo. PMID- 11025193 TI - SV40 T/t-antigens sensitize mammary gland epithelial cells to oxidative stress and apoptosis. AB - As shown recently, the SV40 T/t-antigens (T/t-ag) exert a strong apoptotic activity in mouse mammary gland epithelial cells (ME-cells) leading to premature gland involution at late pregnancy. This high spontaneous cell death rate (20%) is also maintained in T/t-ag positive ME-tissue culture cell lines (e.g., 8/61 A), but not in those ME-cells that have switched off the SV40 T/t-transgene expression. In this study, we demonstrate for the first time that the T/t-ag sensitize ME-cells to oxidative stress leading to apoptosis. Treatment of the 8/61-A ME-cells with catalase, a scavenger of H2O2, completely blocked spontaneous cell death, which was linked to downregulation of caspase-3 activity. Furthermore, exposure of the cells to low concentrations of H2O2 highly increased the apoptosis rate. These findings suggest that the T/t-ag positive ME-cells contain either elevated levels of reactive oxygen species or reduced antioxidant activities. During spontaneous and H2O2-induced apoptosis, the activity of caspase-3 is significantly increased. In addition, the 8/61-A cells accumulated p21 and Bax proteins while the level of the anti-apoptotic protein Bcl-2 decreased implying a posttranscriptional regulation of apoptosis. PMID- 11025194 TI - Mechanism of protection by the flavonoids, quercetin and rutin, against tert butylhydroperoxide- and menadione-induced DNA single strand breaks in Caco-2 cells. AB - Protection by the flavonoids, quercetin and rutin, against tert butylhydroperoxide (tert-BOOH)- and menadione-induced DNA single strand breaks was investigated in Caco-2 cells. Both tert-BOOH and menadione induced DNA single strand breaks in a concentration-dependent manner. Pre-incubation of Caco-2 cells with either quercetin or rutin for 24 h significantly decreased the formation of DNA single strand breaks evoked by tert-BOOH (P <.05). Iron chelators, 1,10 phenanthroline (o-Phen) and deferoxamine mesylate (DFO), also protected against tert-BOOH-induced DNA damage, whereas butylated hydroxytoluene (BHT) had no effect. Quercetin, and not rutin, decreased the extent of menadione-induced DNA single strand breaks. DFO and BHT, and not o-Phen, protected against menadione induced DNA strand break formation (P <.05). From the results of this study, iron ions were involved in tert-BOOH-induced DNA single strand break formation in Caco 2 cells, whereas DNA damage evoked by menadione was far more complex. We demonstrated that the flavonoids, quercetin and rutin, protected against tert BOOH-induced DNA strand breaks by way of their metal ion chelating mechanism. However, quercetin, and not rutin, protected against menadione-induced DNA single strand breaks by acting as both a metal chelator and radical scavenger. PMID- 11025195 TI - Nitric oxide affects the production of reactive oxygen species in hepatoma cells: implications for the process of oxygen sensing. AB - Treatment of human hepatoma cells (HepG2) with NO-donors for 24 h inhibited hypoxia-induced erythropoietin (EPO) gene activation. NO was found to increase the production of reactive oxygen species (ROS), the putative signaling molecules between a cellular O2-sensor and hypoxia inducible factor 1 (HIF-1). HIF-1 is the prime regulator of O2-dependent genes such as EPO. NO-treatment for more than 20 h reduced HIF-1-driven reporter gene activity. In contrast, immediately after the addition of NO, ROS levels in HepG2 cells decreased below control values for as long as 4 h. Corresponding to these lowered ROS-levels, HIF-1 reporter gene activity and EPO gene expression transiently increased but were reduced when ROS levels rose thereafter. Our findings of a bimodal effect of NO on ROS production shed new light on the involvement of ROS in the mechanism of O2-sensing and may explain earlier conflicting data about the effect of NO on O2-dependent gene expression. PMID- 11025196 TI - Nitric oxide interacts with the cAMP pathway to modulate capacitation of human spermatozoa. AB - This study aimed to demonstrate nitric oxide production by human spermatozoa and to characterize the interaction between nitric oxide and cAMP-related pathway in the control of human sperm capacitation and protein tyrosine phosphorylation. Spermatozoa were incubated in Tyrode's medium with or without bovine serum albumin (BSA), and nitric oxide was measured with the spin trap sodium N-methyl-D glucamine dithiocarbamate. Under noncapacitating conditions, spermatozoa produced low levels of nitric oxide. However, under capacitating conditions, prominent nitric oxide adduct signals were obtained and a time-dependent increase of nitric oxide production was observed. When spermatozoa were incubated in Tyrode+BSA medium with nitric oxide-releasing compounds, intracellular cAMP concentrations increased to levels higher than those of spermatozoa incubated in Tyrode+BSA alone. In contrast, incubation with nitric oxide synthase inhibitors (N(G)-nitro L-arginine methyl ester or N(G)-monomethyl L-arginine) decreased intracellular sperm cAMP concentrations. The inhibitory effect observed with N(G)-nitro-L arginine methyl ester on capacitation and tyrosine phosphorylation of two sperm proteins (105, 81 kDa) was overcome by the presence of cAMP analogs or of a phosphodiesterase inhibitor. These results indicate that nitric oxide is produced by capacitating human spermatozoa and that it may act as a cellular messenger by modulating the cAMP pathway involved in capacitation and protein tyrosine phosphorylation. PMID- 11025197 TI - Oxidative stress-induced apoptosis of endothelial cells. AB - Endothelial cells (ECs) are subjected to oxidative stress during many pathological processes, including ischemia/reperfusion and general inflammation. In the present study, we examined the effects of oxidative stress on rates of apoptosis in EC cultures. We treated large and microvessel ECs with menadione for 1 h in vitro to simulate the most common physiological form of oxidative stress, exposure to O2*-. Capillary ECs were resistant to menadione-induced apoptosis when compared with large-vessel ECs. Treatment with 35 microM menadione resulted in an apoptotic rate of approximately 5% in capillary EC cultures compared with approximately 45% in large-vessel EC cultures. At higher concentrations of menadione (35-75 microM), both types of ECs exhibited a concentration-related increase in apoptosis. Necrotic cell death only became evident at menadione concentrations ranging from 75-100 microM for both cell types. The timing of the apoptotic response to a 1 h menadione exposure was very specific. For both EC types, peaks of apoptosis occurred in two distinct waves, at 6-8 and 18-22 h after treatment. Analysis of the events leading up to the first peak of apoptosis indicated that specific matrix metalloproteinases (MMPs) were activated, suggesting that MMPs may be involved in initiating the apoptotic process. PMID- 11025198 TI - Trace detection of hydroxyl radicals during the redox cycling of low concentrations of diaziquone: a new approach. AB - Quantifying oxygen radicals that arise during the redox cycling of quinone containing anticancer agents such as diaziquone (AZQ) has been difficult, as has been their detection at low drug concentrations. This is due to the fact that EPR spin trapping, the method most often used for *OH detection, requires the use of high drug concentrations. Using a new highly sensitive technique that employs a fluorescamine-derivatized nitroxide, we show that low levels of NADPH-cytochrome P450 reductase (4.25 microg/ml) catalyze the production of hydroxyl radicals at very low, clinically relevant AZQ concentrations. Thus, at this enzyme concentration, we were able to detect a rate of 0.10 nM s(-1) hydroxyl radical production by 5 microM AZQ, a clinically relevant concentration. The Michaelis Menten constants for AZQ-mediated hydroxyl radical production are: K(M) = 10.7 +/ 1.4 microM, and V(max) = 5.2 +/- 0.9 x 10(-8) M s(-1) (mg protein)(-1). Experiments employing catalase, superoxide dismutase, and NADPH-cytochrome P450 reductase, confirm the previously deduced conclusions from high drug concentrations, that is, that at low concentrations, AZQ acts to shuttle reducing equivalents from the enzyme to oxygen, thus generating the redox cycle. The data presented here suggest that the levels and locations of redox active metal ions may be the principal controlling factor in the pathway of AZQ activity that involves oxidative stress. PMID- 11025199 TI - Formation of a protein-bound pyrazinium free radical cation during glycation of histone H1. AB - Glycation, the nonenzymatic reaction between protein amino groups and reducing sugars, induces protein damage that has been linked to several pathological conditions, especially diabetes, and general aging. Here we describe the direct identification of a protein-bound free radical formed during early glycation of histone H1 in vitro. Earlier EPR analysis of thermal browning reactions between free amino acids and reducing sugars has implicated the sugar fragmentation product glycolaldehyde in the generation of a 1,4-disubstituted pyrazinium free radical cation. In order to evaluate the potential formation of this radical in vivo, the early glycation of BSA, lysozyme, and histone H1 by several sugars (D glucose, D-ribose, ADP-ribose, glycolaldehyde) under conditions of physiological pH and temperature was examined by EPR. The pyrazinium free radical cation was identified on histone H1 glycated by glycolaldehyde (g = 2.00539, aN = 8.01 [2N], aH = 5.26 [4H], aH = 2.72 [4H]), or ADP-ribose. Reaction of glycoaldehyde with poly-L-lysine produced an identical signal, whereas reaction with BSA or lysozyme produced only a minor unresolved singlet signal. In the absence of oxygen the signal was stable over several days. Our results raise the possibility that pyrazinium radicals may form during glycation of histone H1 in vivo. PMID- 11025200 TI - EPR detection of lipid-derived free radicals from PUFA, LDL, and cell oxidations. AB - We have used the spin trap 5,5-dimethyl-pyrroline-1-oxide (DMPO) and EPR to detect lipid-derived radicals (Ld*) during peroxidation of polyunsaturated fatty acids (PUFA), low-density lipoprotein (LDL), and cells (K-562 and MCF-7). All oxygen-centered radical adducts of DMPO from our oxidizable targets have short lifetimes (<20 min). We hypothesized that the short lifetimes of these spin adducts are due in part to their reaction with radicals formed during lipid peroxidation. We proposed that stopping the lipid peroxidation processes by separating oxidation-mediator from oxidation-substrate with an appropriate extraction would stabilize the spin adducts. To test this hypothesis we used ethyl acetate to extract the lipid-derived radical adducts of DMPO (DMPO/Ld*) from an oxidizing docosahexaenioc acid (DHA) solution; Folch extraction was used for LDL and cell experiments. The lifetimes of DMPO spin adducts post-extraction are much longer (>10 h) than the spin adducts detected without extraction. In iron-mediated DHA oxidation we observed three DMPO adducts in the aqueous phase and two in the organic phase. The aqueous phase contains DMPO/HO* aN approximately aH approximately 14.8 G) and two carbon-centered radical adducts (aN1 approximately 15.8 G, aH1 approximately 22.6 G; aN2 approximately 15.2 G, aH2 approximately 18.9 G). The organic phase contains two long-chain lipid radical adducts (aN approximately 13.5 G, aH approximately 10.2 G; and aN approximately 12.8 G; aH approximately 6.85 G, 1.9 G). We conclude that extraction significantly increases the lifetimes of the spin adducts, allowing detection of a variety of lipid-derived radicals by EPR. PMID- 11025201 TI - Detailed methods for the quantification of nitric oxide in aqueous solutions using either an oxygen monitor or EPR. AB - The interest in nitric oxide has grown with the discovery that it has many biological functions. This has heightened the need for methods to quantify nitric oxide. Here we report two separate methods for the quantification of aqueous stock solutions of nitric oxide. The first is a new method based on the reaction of nitric oxide with oxygen in liquid phase (*NO + O2 + 2H2O --> 4HNO2); an oxygen monitor is used to measure the consumption of oxygen by nitric oxide. This method offers the advantages of being both simple and direct. The presence of nitrite or nitrate, frequent contaminants in nitric oxide stock solutions, does not interfere with the quantification of nitric oxide. Measuring the disappearance of dissolved oxygen, a reactant, in the presence of known amounts of nitric oxide has provided verification of the 4:1 stoichiometry of the reaction. The second method uses electron paramagnetic resonance spectroscopy (EPR) and the nitric oxide trap [Fe2+-(MGD)2], (MGD = N-methyl-D-glucamine dithiocarbamate). The nitrosyl complex is stable and easily quantitated as a room temperature aqueous solution. These two methods are validated with Sievers 280 Nitric Oxide Analyzer and cross-checked with standards using UV-Vis spectroscopy. The practical lower limits for measuring the concentration of nitric oxide using the oxygen monitor approach and EPR are approximately 3 microM and 500 nM, respectively. Both methods provide straightforward approaches for the standardization of nitric oxide in solution. PMID- 11025202 TI - Yeast two-hybrid system identifies the ubiquitin-conjugating enzyme mUbc9 as a potential partner of mouse Dac. AB - Using a yeast two hybrid system and pull-down assays we demonstrate that mouse Dac (mDac) specifically binds to mouse ubiquitin-conjugating enzyme mUbc9. In contrast to a direct interaction between Drosophila dachshund (dac) and eyes absent (eya)gene products, we cannot detect by the same methods that mDac binds to mEya2, a functional mouse homologue of the Drosophila Eya. Immunostaining of various cell lines that were transfected with mDac reveals that mDac protein is found predominantly in the nucleus but translocates to the cytoplasm and condensates along the nuclear membrane in a cell-cycle dependent manner. Deletion analysis of mDac show the intracellular localization and protein stability correlates with the binding to mUbc9. The C-terminal half of mDac, which associates with mUbc9, remains cytoplasmic and is degraded in proteasome whereas the non-interacting N-terminus is exclusively nuclear and more stable than the full-length mDac or its C-terminal portion. In situ hybridization on whole-mount embryos or tissue sections detects mUbc9 transcripts in complementary and overlapping areas with mDac expression, particularly in the proliferation zone of the limb buds, the spinal cord and forebrain. Mouse embryos stained with an anti mDac antibody document that mDac is localized both in the nucleus and the cytoplasm with a cytoplasmic predominance in migrating neural crest cells. In the proliferation zone, visible nuclear envelopes are not formed and mDac is detected throughout the cells. PMID- 11025203 TI - Isolation and characterization of AINT: a novel ARNT interacting protein expressed during murine embryonic development. AB - Basic helix-loop-helix-PER-ARNT-SIM (bHLH-PAS) proteins form dimeric transcription factors to mediate diverse biological functions including xenobiotic metabolism, hypoxic response, circadian rhythm and central nervous system midline development. The Ah receptor nuclear translocator protein (ARNT) plays a central role as a common heterodimerization partner. Herein, we describe a novel, embryonically expressed, ARNT interacting protein (AINT) that may be a member of a larger coiled-coil PAS interacting protein family. The AINT C terminus mediates interaction with the PAS domain of ARNT in yeast and interacts in vitro with ARNT and ARNT2 specifically. AINT localizes to the cytoplasm and overexpression leads to non-nuclear localization of ARNT. A dynamic pattern of AINT mRNA expression during embryogenesis and cerebellum ontogeny supports a role for AINT in development. PMID- 11025204 TI - Ets family transcription factor ESE-1 is expressed in corneal epithelial cells and is involved in their differentiation. AB - Involvement of an epithelium-specific transcription factor ESE-1/ESX/ELF3/jen (ESE-1) in corneal epithelial cell differentiation was investigated. ESE-1 was reported to be induced during terminal differentiation of the epidermis and primary keratinocytes and to transactivate target genes through ets binding sites. However, its expression and function in corneal epithelium have not been examined. We report here that ESE-1 is upregulated upon differentiation in mouse corneal epithelium and in immortalized human corneal epithelial cells (HCE). ESE 1 transactivates through the regulatory element of cornea-specific K12 keratin. Moreover, introduction of ESE-1 antisense RNA in HCE cells affect their differentiation. These data suggest the involvement of ESE-1 in differentiation of corneal epithelial cells. PMID- 11025205 TI - Repulsive axon guidance molecule Sema3A inhibits branching morphogenesis of fetal mouse lung. AB - Semaphorin III/collapsin-1 (Sema3A) guides a specific subset of neuronal growth cones as a repulsive molecule. In this study, we have investigated a possible role of non-neuronal Sema3A in lung morphogenesis. Expression of mRNAs of Sema3A and neuropilin-1 (NP-1), a Sema3A receptor, was detected in fetal and adult lungs. Sema3A-immunoreactive cells were found in airway and alveolar epithelial cells of the fetal and adult lungs. Immunoreactivity for NP-1 was seen in fetal and adult alveolar epithelial cells as well as endothelial cells. Immunoreactivity of collapsin response mediator protein CRMP (CRMP-2), an intracellular protein mediating Sema3A signaling, was localized in alveolar epithelial cells, nerve tissue and airway neuroendocrine cells. The expression of CRMP-2 increased during the fetal, neonate and adult periods, and this pattern paralleled that of NP-1. In a two-day culture of lung explants from fetal mouse lung (E11.5), with exogenous Sema3A at a dose comparable to that which induces growth cone collapse of dorsal root ganglia neurons, the number of terminal buds was reduced in a dose-dependent manner when compared with control or untreated lung explants. This decrease was not accompanied with any alteration of the bromodeoxyuridine-positive DNA-synthesizing fraction. A soluble NP-1 lacking the transmembrane and intracellular region, neutralized the inhibitory effect of Sema3A. The fetal lung explants from neuropilin-1 homozygous null mice grew normally in vitro regardless of Sema3A treatment. These results provide evidence that Sema3A inhibits branching morphogenesis in lung bud organ cultures via NP-1 as a receptor or a component of a possible multimeric Sema3A receptor complex. PMID- 11025206 TI - A novel function for the Xslug gene: control of dorsal mesendoderm development by repressing BMP-4. AB - The Snail family of genes comprise a group of transcription factors with characteristic zinc finger motifs. One of the members of this family is the Slug gene. Slug has been implicated in the development of neural crest in chick and Xenopus by antisense loss of function experiments. Here, we have generated functional derivatives of Xslug by constructing cDNAs that encode the Xslug protein fused with the transactivation domain of the virus-derived VP16 activator or with the repressor domain of the Drosophila Engrailed protein. Our results suggest that Xslug normally functions as a transcriptional repressor and that Xslug-VP16 behaves as a dominant negative of Xslug. In the present work, we confirm and extend previous results that suggest that Xslug has an important function in neural crest development, by controlling its own transcription. In addition we have uncovered a new function for Xslug. We show that Xslug is expressed in the dorsal mesendoderm at the beginning of gastrulation, where is it able to upregulate the expression of dorsal genes. On the other hand when Xslug is expressed outside of the organizer it represses the expression of ventral genes. Our results indicate that this effect on mesodermal patterning depends on BMP activity, showing that Xslug can directly control the transcription of BMP-4. PMID- 11025207 TI - The Drosophila homolog of Onecut homeodomain proteins is a neural-specific transcriptional activator with a potential role in regulating neural differentiation. AB - We report here the characterization of the Drosophila homolog of the onecut homeobox gene, which encodes a protein product with one cut domain and one homeodomain. We present evidence that D-Onecut can bind to similar DNA sequences with high specificity and affinity as other Onecut proteins through the highly conserved cut domain and homeodomain. Interestingly, the cut domain alone can mediate DNA-binding, but the homeodomain cannot. However, depending upon the promoter context, we observed cooperative interactions between the two domains to confer high DNA-binding affinity and specificity. D-Onecut appears to be a moderate transcriptional activator and functions as a nuclear protein in neuronal tissues of both the CNS and PNS during development and in the adult. In the eye, D-Onecut expression is independent of glass, a transcriptional regulator of R cell differentiation. Taken together, our results suggest a role for D-Onecut in the regulation of some aspects of neural differentiation or maintenance. In support of this notion, overexpression of a putative dominant negative form of D Onecut during eye development does not affect early cell fate specification, but severely affects photoreceptor differentiation. PMID- 11025208 TI - Expression of the cyclin-dependent kinase inhibitor Dacapo is regulated by cyclin E. AB - The Cip/Kip family of cyclin-dependent kinase inhibitors (CKIs) has been implicated in mediating cell cycle arrest prior to terminal differentiation. In many instances, increased expression of CKIs immediately precedes mitotic arrest. However, the mechanism that activates CKI expression in cells that are about to stop dividing has remained elusive. Here we have addressed this issue by investigating the expression pattern of dacapo, a Cip/Kip CKI in Drosophila. We show that the accumulation of dacapo RNA and protein requires Cyclin E and that increased expression of Cyclin E can induce dacapo expression. We also show that the oscillation of the Cyclin E and Dacapo proteins are tightly coupled during ovarian endocycles. Our results argue for a mechanism where Cyclin E/Cdk activity induces Dacapo expression but only within certain windows that are permissive for dacapo expression. PMID- 11025209 TI - The Toll/IL-1 receptor binding protein MyD88 is required for Xenopus axis formation. AB - The Toll/Dorsal pathway regulates dorsoventral axis formation in the Drosophila embryo. We had previously obtained evidence that a homologous pathway exists in Xenopus, however, its role during normal frog development had not been established. Here we report the cloning of Xenopus MyD88 (XMyD88), whose mammalian homologs are adaptor proteins linking Toll/IL-1 receptors and IRAK kinases. We show that in the frog embryo overexpression of a dominant-negative form of XMyD88 blocked Toll receptor activity, specifically inhibited axis formation and reduced expression of pivotal organizer genes. The observed stage dependency of interference suggests a function for maternal XMyD88 soon after fertilization. We conclude that XMyD88 activity is required for normal Spemann organizer formation, implying an essential role for maternal Toll/IL-1 receptors in Xenopus axis formation. PMID- 11025211 TI - Left-right asymmetry in BMP4 signalling pathway during chick gastrulation. AB - Determination of the left-right (L-R) axis was shown recently to implicate several genes, among which TGFbeta-related molecules such as Activin betaB, lefty1 and 2 and Nodal. We show here that Bmp4 and its signal transduction pathway partners BMPR IA and Smad1 are transiently expressed on the right side of Hensen's node, when L-R polarity is being established. Moreover, Smad1 is expressed asymmetrically in the nascent notochord. These observations suggest a role for a BMP4-dependent autocrine or paracrine mechanism during early L-R determination. PMID- 11025210 TI - Differentiation of the epithelial apical junctional complex during mouse preimplantation development: a role for rab13 in the early maturation of the tight junction. AB - We have investigated the mechanisms by which the epithelial apicolateral junctional complex (AJC) is generated during trophectoderm differentiation in the mouse blastocyst using molecular, structural and functional analyses. The mature AJC comprises an apical tight junction (TJ), responsible for intercellular sealing and blastocoel formation, and subjacent zonula adherens E cadherin/catenin adhesion complex which also extends along lateral membrane contact sites. Dual labelling confocal microscopy revealed that the AJC derived from a single 'intermediate' complex formed following embryo compaction at the 8 cell stage in which the TJ-associated peripheral membrane protein, ZO-1alpha- isoform, was co-localized with both alpha- and beta-catenin. However, following assembly of the TJ transmembrane protein, occludin, from the early 32-cell stage when blastocoel formation begins, ZO-1alpha- and other TJ proteins (ZO-1alpha+ isoform, occludin, cingulin) co-localized in an apical TJ which was separate from a subjacent E-cadherin/catenin zonula adherens complex. Thin-section electron microscopy confirmed that a single zonula adherens-like junctional complex present at the AJC site following compaction matured into a dual TJ and zonula adherens complex at the blastocyst stage. Embryo incubation in the tracer FITC dextran 4 kDa showed that a functional TJ seal was established coincident with blastocoel formation. We also found that rab13, a small GTPase previously localized to the TJ, is expressed at all stages of preimplantation development and relocates from the cytoplasm to the site of AJC biogenesis from compaction onwards with rab13 and ZO-1alpha- co-localizing precisely. Our data indicate that the segregation of the two elements of the AJC occurs late in trophectoderm differentiation and likely has functional importance in blastocyst formation. Moreover, we propose a role for rab13 in the specification of the AJC site and the formation and segregation of the TJ. PMID- 11025212 TI - mTert expression correlates with telomerase activity during the differentiation of murine embryonic stem cells. AB - Telomerase, the enzyme which maintains the ends of linear chromosomes in eukaryotic cells, is found at low levels in somatic stem cells but while this is incapable of preventing the progressive erosion of telomeres occurring as a consequence of cell division, such cells show greater proliferative capacity than normal somatic cells hence examination of telomerase activity in such stem cells is of interest. Our aim in this work was to examine the relationship between expression of the reverse transcriptase component (mTert) of murine telomerase. We report here the insertion of a reporter cassette comprising a segment of the promoter sequence of murine Tert gene coupled to the coding sequence of green fluorescent protein (GFP) into murine embryonic stem (ES) cells and show that this is sufficient for mimicking the expression of mTert. We show that the expression of mTert is very closely linked to telomerase activity and that both are substantially reduced upon differentiation of ES cells into more committed lineages giving us a potential reporter system for the selection and isolation of ES cells possessing different levels of telomerase activity. PMID- 11025213 TI - Bottle cell formation in relation to mesodermal patterning in the Xenopus embryo. AB - The appearance of bottle cells at the dorsal vegetal/marginal boundary of Xenopus embryos marks the onset of blastopore formation. The conditions leading to this epithelial activity were investigated by inducing bottle cells ectopically in the animal region with VegT or different members of the transforming growth factor (TGF)-beta family. Morphological studies on the ectopic bottle cells indicate their close similarity to the endogenous bottle cells at the dorsal blastopore lip. The subepithelial cells of the induced animal region express mesodermal genes in a pattern reminiscent to that observed on the dorsal lip. Relating this expression pattern to the position of the ectopic bottle cells leads to the conclusion that bottle cells form in regions of high TGF-beta signalling. The specific inhibitory effects of cerberus on ectopically induced bottle cells revealed that nodal related growth factors are the intrinsic signals that elicit bottle cell formation in the normal embryo. In addition, fibroblast growth factor signalling is an essential precondition for this epithelial response as it is for mesoderm formation. We conclude that bottle cell formation in the epithelial layer of the gastrula is closely linked to mesodermal patterning in the subepithelial tissues. PMID- 11025214 TI - A genetic screen for mutations affecting embryonic development in medaka fish (Oryzias latipes). AB - In a pilot screen, we assayed the efficiency of ethylnitrosourea (ENU) as a chemical mutagen to induce mutations that lead to early embryonic and larval lethal phenotypes in the Japanese medaka fish, Oryzias latipes. ENU acts as a very efficient mutagen inducing mutations at high rates in germ cells. Three repeated treatments of male fish in 3 mM ENU for 1 h results in locus specific mutation rates of 1.1-1.95 x10(-3). Mutagenized males were outcrossed to wild type females and the F1 offspring was used to establish F2 families. F2 siblings were intercrossed and the F3 progeny was scored 24, 48 and 72 h after fertilization for morphological alterations affecting eye development. The presented mutant phenotypes were identified using morphological criteria and occur during early developmental stages of medaka. They are stably inherited in a Mendelian fashion. The high efficiency of ENU to induce mutations in this pilot screen indicates that chemical mutagenesis and screening for morphologically visible phenotypes in medaka fish allows the genetic analysis of specific aspects of vertebrate development complementing the screens performed in other vertebrate model systems. PMID- 11025215 TI - Expression of the bZIP transcription factor TCF11 and its potential dimerization partners during development. AB - TCF11 (also known as Nrf1 and LCR-F1) is a basic-region leucine-zipper (b-ZIP) transcription factor that is essential during embryonic development. We have carried out expression analysis at a number of developmental stages and find that while there is some localized elevated expression between 8 and 9 days post coitum (dpc), the gene is widely expressed with a constant level of mRNA transcripts detectable in all tissues and at all stages examined. However, this does not reflect the specific nature of a TCF11 mutant phenotype (EMBO J. 17 (1998) 1779) which shows an essential role in foetal liver haematopoiesis. The specificity of TCF11 function may be controlled at a post-transcriptional level including availability of, and specific interaction with, a range of potential heterodimerization partners. We therefore carried out expression analysis of four candidate partner molecules; the three small Maf genes and another bZIP transcription factor found to bind to TCF11 in a two-hybrid screen, ATF4. We show different patterns of expression for the three Maf genes during development with MafG being widely expressed, MafK expressed only later in development and in specific tissues, and no detection of MafF. ATF4 shows evidence of complex regulation during development and shows elevated expression in many of the same sites as TCF11. PMID- 11025216 TI - Dynamic expression and activity of NF-kappaB during post-natal mammary gland morphogenesis. AB - The Rel/NF-kappaB family of transcription factors has been implicated in such diverse cellular processes as proliferation, differentiation, and apoptosis. As each of these processes occurs during post-natal mammary gland morphogenesis, the expression and activity of NF-kappaB factors in the murine mammary gland were examined. Immunohistochemical and immunoblot analyses revealed expression of the p105/p50 and RelA subunits of NF-kappaB, as well as the major inhibitor, IkappaBalpha, in the mammary epithelium during pregnancy, lactation, and involution. Electrophoretic mobility shift assay (EMSA) demonstrated that DNA binding complexes containing p50 and RelA were abundant during pregnancy and involution, but not during lactation. Activity of an NF-kappaB-dependent luciferase reporter in transgenic mice was highest during pregnancy, decreased to near undetectable levels during lactation, and was elevated during involution. This highly regulated pattern of activity was consistent with the modulated expression of p105/p50, RelA, and IkappaBalpha. PMID- 11025217 TI - Fhx (Foxj2) expression is activated during spermatogenesis and very early in embryonic development. AB - FHX (FOXJ2) is a recently characterized human fork head transcriptional activator that binds DNA with a dual sequence specificity. We have cloned the cDNA for the mouse orthologue Foxj2 and characterized its expression in the gonads and along the early pre-implantation development of the mouse. In the testis, Foxj2 is expressed from pachytene spermatocytes to round spermatids, but not in spermatogonia. In addition to the germ lineage, only Sertoli cells of the testis showed expression of Foxj2. In the ovary, only granulosa cells of the follicles express the factor. Neither mature spermatozoa nor oocytes showed expression of Foxj2. Foxj2 expression is early activated in zygotic development, being detected since as early as 8-cell stage embryos. Both cell layers of the blastocyst: the trophectoderm (TE) and the inner cell mass (ICM), express Foxj2. PMID- 11025218 TI - Inner ear and lateral line expression of a zebrafish Nkx5-1 gene and its downregulation in the ears of FGF8 mutant, ace. AB - An orthologue of the mouse homeobox gene Nkx5-1 was cloned and characterized in the zebrafish. As in the mouse and chick, the zebrafish Nkx5-1 gene is expressed in the ear placode and vesicle and in cells forming the vestibulo-acoustic ganglion. In addition, a novel expression domain, the lateral line, appears in the zebrafish, supporting a common precursor hypothesis for these two organs. In the FGF8 zebrafish mutant ace, expression of Nkx5-1 in the otic structures is diminished. The most significant reduction of zfNkx5-1 expression was observed in cells of the vestibulo-acoustic ganglion. PMID- 11025219 TI - Expression of the follistatin/EGF-containing transmembrane protein M7365 (tomoregulin-1) during mouse development. AB - A novel transmembrane protein (designated X7365) containing two follistatin modules and an epidermal growth factor (EGF) domain has been described in the hypothalamic-pituitary axis of Xenopus laevis. We have now cloned the highly conserved mouse orthologue (M7365), and its mRNA was detected in many mesodermal and (neuro)ectodermal tissues in 8.5-day-old mouse embryos. During further development, M7365 mRNA expression became restricted to certain regions in the brain and to ganglia. In the adult mouse, the brain is the major site of M7365 expression. PMID- 11025220 TI - Expression of the Kruppel-like zinc finger gene biklf during zebrafish development. AB - The zebrafish biklf gene encodes a novel Kruppel-like transcription factor containing three contiguous zinc fingers at the C-terminus. Expression of biklf is detected from the shield stage onward in the developing prechordal plate, and as a 'baseball seam'-like lateral stripe beginning at the end of gastrulation. The latter expression domain is suppressed in the swirl mutant in which bmp2b is disrupted. At the 5-somite stage the lateral expression domain separates into two distinct stripes, one in the ectoderm, the other in blood islands in the lateral plate mesoderm. Blood island staining of biklf continues through somitogenesis as the most prominent area of biklf expression. PMID- 11025221 TI - Sfrp-1 and sfrp-2 are expressed in overlapping and distinct domains during chick development. AB - Secreted frizzled related proteins (Sfrps) are thought to bind and regulate Wnt activity through a cysteine rich domain that is highly similar to that of Frizzled receptors. To investigate possible roles for Sfrps in chick development, we have isolated partial cDNAs encoding Sfrp-1 and Sfrp-2 and have thoroughly characterized the expression patterns of both genes. Both sfrp-1 and sfrp-2 are expressed at all stages of development analyzed, ranging from Hamburger and Hamilton stage 4 through stage 32. Expression of both sfrp-1 and sfrp-2 is observed in mesodermal and ectodermal derivatives, while sfrp-1 is also found in endodermal lineages. PMID- 11025223 TI - Xenopus cadherin-6 is expressed in the central and peripheral nervous system and in neurogenic placodes. AB - We have cloned the cDNAs encoding the two Xenopus laevis cadherin-6 pseudoalleles (Xcad-6 and Xcad-6'), which belong to the classical type II cadherin subfamily. During embryogenesis, Xcad-6 expression starts at early neurula and increases dramatically until tadpole stages. Whole-mount in situ hybridization revealed that the transcripts are localized in the developing brain and eye. However, most prominent expression was found in the peripheral nervous system, including a major proportion of lateral line structures. Our results implicate an important contribution of cadherin-6 to the formation of placode derived nerves in Xenopus, an observation which has not been described for other species. PMID- 11025222 TI - The zebrafish scyba gene encodes a novel CXC-type chemokine with distinctive expression patterns in the vestibulo-acoustic system during embryogenesis. AB - Chemokines, in addition to their characterized functions as immune modulators, also play a role in developmental processes such as neural cell migration. Although, chemokines have been described in human, mouse and other vertebrate species, they have yet to be characterized in zebrafish. In this paper, we report the isolation and expression analysis of scyba, a zebrafish gene encoding a CXC type chemokine protein. During early segmentation, scyba transcripts are detected in the midbrain region and the otic placodes. At later developmental stages, scyba expression is restricted to a subset of hindbrain commissural neurons and to the hair-cell sensory patches of the otic vesicle and lateral-line neuromasts. PMID- 11025224 TI - Expression of the zinc finger gene fez-like in zebrafish forebrain. AB - Anterior-posterior (A-P) patterning in the neuroectoderm is established during gastrulation in zebrafish and amphibians. We isolated a novel zinc-finger gene fez-like (fezl) from zebrafish, which displays sequence similarities to Xenopus Fez. The fezl transcripts were detected in the anterior edge of neuroectoderm, the prospective dorsal forebrain, from the late gastrula (80% epiboly stage) to the mid-segmentation period. fezl was also expressed in the ventral forebrain overlying the prechordal plate at these stages. The expression of fezl was enhanced in embryos expressing the Wnt inhibitor Dkk1 and reduced in embryos expressing Wnt8b. The expression in the ventral forebrain was eliminated in the one-eyed pinhead mutant and the antivin RNA-injected embryos, which lack the prechordal plate. Radiation hybrid mapping revealed that the fezl gene is localized to linkage group 11. PMID- 11025225 TI - Developmental expression of the estrogen receptor-related receptor gamma in the nervous system during mouse embryogenesis. AB - The ERR's (estrogen receptor-related receptors) are constitutive activators of the classical estrogen response element. In this report, we demonstrate that ERRgamma is highly expressed in the nervous system of the developing mouse embryo and that the adult pattern of expression of ERRgamma is, with few exceptions, established during embryogenesis. Transcripts are preferentially detected in already differentiating areas of the nervous system. PMID- 11025227 TI - Nerfin-1 and -2, novel Drosophila Zn-finger transcription factor genes expressed in the developing nervous system. AB - To gain insight into the regulatory networks controlling Drosophila neural identity decisions, we have identified new neuronal precursor genes by performing an in situ hybridization screen of differentially selected embryonic head cDNAs. Here, we describe the molecular characteristics and expression profile of nerfin 1, a novel pan-neural precursor gene. This paper also documents the embryonic expression of another structurally related gene, nerfin-2. During early CNS development, nerfin-1 gene expression is activated in neuroblasts (NBs) prior to lineage formation. However, after early sublineage development, nerfin-1 expression shifts from NBs to ganglion mother cells (GMCs) but is not expressed in neurons or glia. Differing from nerfin-1, nerfin-2 is expressed only in a subset of brain neurons. Possessing a conserved putative DNA-binding domain, the predicted Nerfin-1 and -2 proteins belong to a subfamily of Zn-finger transcription factors with cognates identified in nematode, mouse and man. PMID- 11025226 TI - Tissue-specific expression of GTPas RalA and RalB during embryogenesis and regulation by epithelial-mesenchymal interaction. AB - RalA and RalB are members of the Ras-like GTP-binding protein family. To investigate if Ral GTPases play a role in development, temporal and spatial patterns of RalA and RalB gene expression were examined during embryogenesis. RalA and RalB had tissue-specific pattern of expression in developing organs. Both GTPases were expressed in endodermal derivatives in the digestive tract, kidney, mesonephros, gonad, adrenal gland, trachea, and blood vessels. RalA and RalB were also expressed in the ectodermal derivatives in the brain, spinal cord, ganglia, eye, skin, hair, whiskers, and teeth. RalA and RalB were also expressed in some mesodermal tissues including the lung mesenchyme and myocardium. Tissue recombination experiments showed that of Ral expression was increased by epithelial-mesenchymal interaction in the lung, indicating that Ral expression is functionally regulated. These data identify that RalA and RalB as inducible GTPases in the developing embryo. PMID- 11025228 TI - E2F transcription factors are differentially expressed in murine gametes and early embryos. AB - We have examined the murine genes encoding transcription factors E2F1, -3, -5 and -6 in gametes and early embryos. All genes are expressed as maternal transcripts and all are efficiently transcribed after the blastocyst stage. Between those two stages, each E2F mRNA is transcribed with a distinctive and unique pattern. E2F proteins are also differentially expressed and compartmentalized in pre implantation embryos. PMID- 11025229 TI - Early and dynamic expression of cSfrp1 during chick embryo development. AB - Secreted frizzled related proteins (SFRPs) are a new class of signalling molecules that appear to antagonise the activity of the Wnt proteins. Here we report the dynamic expression pattern of cSfrp1, a new member of this family, at early stages of chick embryo development. cSfrp1 transcripts are first detected at pre-streak stages throughout the chick blastula but, during early primitive streak formation, expression is restricted to the anterior primitive streak and later to the blastoderm anterior to the Hensen' s node. This pattern of expression overlaps with that of Otx2 and is complementary to that of cWnt8c. During neural plate formation cSfrp1 mRNAs are abundantly localized only to the anterior domain of the embryo but, as neural tube closes, the expression extends caudally. Later, the main sites of expression in the neural tissue are the telencephalic vesicles, the epiphysis, the developing eyes and the ventral hindbrain and neural tube. Additionally, cSfrp1 transcripts were found in the axial and lateral mesoderm, the otic placode, the trigeminal ganglia, the mesoderm of the branchial arches, the developing limb buds, as well as in the mesodermal component of the developing kidney. PMID- 11025230 TI - Expression of the novel basic-helix-loop-helix transcription factor cMespo in presomitic mesoderm of chicken embryos. AB - We have identified a novel chicken gene, cMespo, which encodes a basic-helix-loop helix (bHLH) protein with sequence homology to a subgroup of bHLH transcription factors that have been implicated in somitogenesis. cMespo transcripts are first found in the primitive streak of gastrulating chick embryos (HH stage 4) and continue to accumulate in presomitic mesoderm (PSM) until somite formation has been concluded. cMespo, however, is not expressed within somites or in tailbud mesoderm. The expression domain of cMespo in PSM largely overlaps with delta-1 but spares a region of several prospective somites at the rostral end of PSM in which c-Meso and Cek-8 are expressed. PMID- 11025231 TI - RALDH3, a retinaldehyde dehydrogenase that generates retinoic acid, is expressed in the ventral retina, otic vesicle and olfactory pit during mouse development. AB - The enzymes that generate retinoic acid during development have been identified as members of the aldehyde dehydrogenase (ALDH) family. The developmental expression patterns of two ALDHs that function as retinaldehyde dehydrogenases, RALDH1 and RALDH2, have been described. Here we report the cloning and expression of a third retinaldehyde dehydrogenase from the mouse called RALDH3 that shares 94% amino acid sequence identity to a human retinaldehyde dehydrogenase previously named ALDH6. In mouse embryos, RALDH3 expression is first noticed in the ventral optic eminence at E8.75, then in the optic vesicle/cup, otic vesicle, and olfactory placode/pit from E9.5 to E11.5. Expression in the developing eye is primarily localized in the ventral retina, thus indicating that RALDH3 represents the V1 dehydrogenase activity described there earlier. From E8.5 to E10.5 RALDH3 expression is distinct from that of RALDH1 or RALDH2, thus indicating a unique role in sensory organ development. PMID- 11025232 TI - Preoperative clonidine attenuates stress response during emergence from anesthesia. AB - STUDY OBJECTIVES: To investigate whether a single preoperative IV dose of clonidine blunts the hemodynamic and hyperadrenergic responses not only to intubation, but also to extubation. DESIGN: Randomized, double-blind, placebo controlled study. PATIENTS: 29 ASA physical status I and II patients (ages 18-65) who were scheduled for noncardiac, elective surgery. Patients were randomly assigned to either receive clonidine 3 microg/kg IV immediately before anesthesia induction or placebo. INTERVENTIONS: Insertion of a 14 G cannula in a large cubital vein for the determination of plasma catecholamines using local anesthesia. Insertion of a radial artery catheter for measuring blood pressure (BP) using local anesthesia. Transthoracic echocardiography to determine cardiac output (CO). MEASUREMENTS: Heart rate (HR), mean arterial pressure (MAP), CO, and plasma catecholamine concentrations. Measurements were performed: before induction (baseline), during intubation, 10 min after intubation, after surgery, during extubation, and 10 min after extubation. MAIN RESULTS: During intubation MAP, HR, and CO were lower in the clonidine group. Compared with baseline measurements, MAP and CO increased less in the clonidine group during intubation. During extubation, MAP was lower in the clonidine group. CO and MAP increased less as compared with baseline measurements in the clonidine group. Compared with the measurements after surgery CO less in the clonidine group during extubation (p < 0.05 for all results). CONCLUSIONS: A single preoperative IV dose of clonidine (3 microg/kg) blunts the hemodynamic responses due to extubation in noncardiac surgery of intermediate duration. PMID- 11025233 TI - Redefining the preoperative evaluation process and the role of the anesthesiologist. AB - STUDY OBJECTIVE: To assess the effects of implementing an ambulatory and same-day surgery preoperative evaluation patient triage system over a 3-year period. DESIGN: Retrospective analysis of 63,941 ambulatory surgical patients presenting for elective surgery. SETTING: Tertiary care, academic medical institution. INTERVENTIONS: The following preoperative evaluation model components were implemented over a 3-year period: HealthQuest, which is an outpatient preoperative assessment computer program developed by the Department of General Anesthesiology; a general internal medicine clinic designated specifically for preoperative evaluation and medical optimization; disease specific algorithms for both preoperative patient assessment and management; and a preoperative anesthesia clinic that no longer performs preoperative medical optimization. MEASUREMENTS AND MAIN RESULTS: During the 3-year study period ambulatory and same day surgical case volume increased 34.7%. A total of 50,967 patients used HealthQuest as part of their preoperative evaluation. Of these patients 22,744 (35.6%) did not need to see an anesthesiologist until the day of surgery as guided by both a computer-assigned HealthQuest score and surgical classification scheme. Also, 41,197 patients were evaluated in our anesthesia preoperative clinic with a cost per evaluation of $24.86, which increased only 0.9% per year. In addition, both patient interview time and patient dissatisfaction with the preoperative process decreased over the 3-year period. There were 20, 088 patient encounters in the general internal medicine clinic for patient medical evaluation and optimization. The average monthly preoperative surgical delay rate decreased 49% during the study period. Finally, significant monetary saving resulted due to decreased unnecessary laboratory testing. CONCLUSIONS: Efficient, cost-effective patient care can be provided by using this preoperative evaluation model. Some institutions may find portions of this preoperative model applicable to their current situation. PMID- 11025234 TI - Low-dose intrathecal clonidine combined with sufentanil as analgesic drugs in abdominal gynecological surgery. AB - STUDY OBJECTIVES: To determine whether a low dose of spinal clonidine either alone or combined with sufentanil would provide effective analgesia following abdominal surgery, as a supplement to bupivacaine spinal anesthesia. DESIGN: Randomized double-blind study. SETTING: Gynecological surgery, teaching hospital. PATIENTS: 73 ASA physical status I and II patients undergoing gynecological abdominal surgery with spinal anesthesia. INTERVENTIONS: Patients were randomly assigned to one of four groups and prospectively studied to examine anesthesia, analgesia, and adverse effects. The control group received saline as the test drug; the sufentanil group received 10 microg of sufentanil; the clonidine group received 30 microg of clonidine; and the sufentanil/clonidine group received 5 microg of sufentanil plus 15 microg of clonidine. All groups received intrathecal 15 mg of bupivacaine (3 mL) plus the intrathecal test drug (2 mL). The concept of visual analog scale (VAS) was introduced. All patients were premedicated with intravenous midazolam. Rescue analgesics were available. MEASUREMENTS AND MAIN RESULTS: The groups were demographically the same. Sensory block to pinprick at 10 min was higher for clonidine and sufentanil/clonidine groups compared to the control group (p < 0.02). Anesthetic time (Bromage score 2) was also longer for clonidine and sufentanil/clonidine groups compared to the control and sufentanil groups (p < 0.05). Time to first rescue analgesics was shorter in the control group compared to the other groups (p < 0.02). The number of IM diclofenac dose injections in 24 hours was higher in the control group compared to all other groups (p < 0.05). The incidence of adverse effects and ephedrine consumption were similar among groups. CONCLUSIONS: Intrathecal 15- and 30-microg clonidine doses expanded the anesthesia sensory block and duration of motor block, and provided analgesia. PMID- 11025235 TI - Prostaglandin E(1) does not influence plasmatic coagulation, hepatic synthesis, or postoperative blood loss in patients after coronary-artery bypass grafting. AB - STUDY OBJECTIVE: To assess whether postoperatively administered prostaglandin E1 (PGE1) might prevent bleeding in patients after coronary artery bypass grafting (CABG). DESIGN: Prospective, randomized, placebo-controlled trial. SETTING: University-affiliated hospital. PATIENTS: 49 patients scheduled for elective CABG surgery. INTERVENTIONS: The PGE1 group received intravenous PGE(1) up to 15 ng/kg/min for 72 hours after surgery, whereas the placebo group received isotonic saline for the same time period. MEASUREMENTS AND MAIN RESULTS: Nine patients (4 in the PGE1 group vs. 5 in the placebo group) had to be excluded because of hemodynamic instability, and 1 in the placebo group because of gastric bleeding. In the remaining 39 patients (20 vs. 19), no significant differences with regard to hemoglobin levels or platelet count could be observed. There was no significant difference between the groups concerning the amount of packed red blood cells, platelet concentrates, or fresh frozen plasma transfused. No significant differences could be observed regarding laboratory markers of coagulation activation or hepatic synthesis either. CONCLUSIONS: PGE1 did not prevent coagulation disturbances and blood loss when administered postoperatively in patients undergoing CABG. The absence of these expected effects might be explained by the concomitant administration of acetylsalicylic acid, whose antiaggregatory acivity seems to exceed the effects of PGE1. PMID- 11025236 TI - Effect of xenon on endotracheal tube cuff. AB - STUDY DESIGN: To investigate the effect of xenon on the endotracheal tube cuff in comparison to that of nitrous oxide. DESIGN: Prospective, randomized study. SETTING: Laboratory in vitro testing of endotracheal tubes. INTERVENTIONS: Exposure of air-filled endotracheal tube cuffs to xenon or nitrous oxide (79 vol%) mixed with a balance of oxygen (21 vol%) for 3 hours. MEASUREMENTS: The intra-cuff pressure and gas volume in the cuff were measured for 3 hours. MAIN RESULTS AND CONCLUSION: Increase in pressure and gas volume was significantly less and slower with xenon than with nitrous oxide. More nitrous oxide passes into the cuff than xenon. PMID- 11025237 TI - The influence of preemptive spinal anesthesia on postoperative pain. AB - STUDY OBJECTIVE: To examine the influence of spinal anesthesia on postoperative pain and postoperative opioid requirements. DESIGN: Prospective randomized study. SETTING: Bnai-Zion Medical Center, Haifa, Israel-a government hospital. MEASUREMENTS AND MAIN RESULTS: 30 ASA physical status I and II unpremedicated women undergoing elective total abdominal hysterectomy were randomly allocated into two groups of 15 patients each using a sealed envelope technique. Patients in Group 1 were given a subarachnoid injection of 12 mg hyperbaric bupivacaine and after 10 minutes general anesthesia was induced. Patients in Group 2 received only general anesthesia. Anesthesia was induced with midazolam and maintained with oxygen, N2O, isoflurane, and pancuronium. No opioids were given intraoperatively. Postoperatively patient-controlled analgesia (PCA) with morphine was initiated in both groups (1 mg x mL(-1), bolus dose 1 mg, lockout interval 10 minutes, and background infusion 1 mg x mL(-1)) at patient first request for analgesic. Pain was assessed over 24 hours by cumulative morphine dose and visual analog score (VAS). Postoperative PCA morphine consumption at 2, 6, and 24 hours following patient first request for analgesic for Groups 1 and 2 were: 3.1 +/- 1 mg versus 7.2 +/- 3 mg (p = 0.04), 13.4 +/- 2 mg versus 17.2 +/- 4 mg (p = 0.03) and 35.9 +/- 8 mg versus 47.7 +/- 8 mg in Group 2 (p = 0.04). VAS scores at 4, 6, 12, and 24 hours postoperatively were not significantly different between the two groups. CONCLUSIONS: Preoperative neural blockade may reduce postoperative analgesic requirements. PMID- 11025238 TI - Hemodynamic effects of simultaneous administration of intravenous ephedrine and spinal anesthesia for cesarean delivery. AB - STUDY OBJECTIVE: To evaluate the hemodynamic effects of an intravenous (IV) ephedrine bolus given simultaneously with spinal anesthesia for cesarean delivery. DESIGN: Randomized, prospective, double-blind study. SETTING: Tertiary care academic medical center. PATIENTS: 40 ASA physical status I and II nonlaboring women undergoing elective cesarean delivery of term, uncomplicated, singleton pregnancies. INTERVENTIONS: After a 10 mL/kg IV lactated Ringers bolus given over 15 minutes, patients simultaneously received an IV dose of 2 mL (10 mg) ephedrine or 2 mL saline, and an intrathecal dose of 12 mg (1.6 mL 0.75%) bupivacaine with 10 microg of fentanyl. Hypotension, defined as a 20% decrease in mean arterial pressure (MAP), was treated with 10-mg IV doses of ephedrine. MEASUREMENTS: Hemodynamic (heart rate and MAP) and non-invasive thoracic impedance (cardiac index, stroke index, and systemic vascular resistance index) measurements at baseline and at 1-minute intervals until the time of delivery. Total fluid, supplemental ephedrine, and Apgar scores were recorded. Data were evaluated using analysis of variance (ANOVA), ANOVA for repeated measures, and Mann-Whitney U-tests where appropriate, with p < 0.05 considered significant. MAIN RESULTS: Significant changes from baseline in MAP, systemic vascular resistance index, heart rate, and cardiac index were observed in both groups after spinal anesthesia. However, there were no differences between the ephedrine and saline groups in the incidence and severity of change from the baseline. The overall incidence of hypotension was 70% in both groups. No difference in total fluid or supplemental ephedrine use was observed. CONCLUSIONS: 10 mg of IV ephedrine given at the time of spinal anesthesia, and after a 10 mL/kg lactated Ringers fluid bolus, does not diminish the incidence or severity of hypotension in parturients undergoing cesarean delivery. PMID- 11025239 TI - Inhibition of the cortisol response to surgical stress in chronically depressed patients. AB - STUDY OBJECTIVE: To evaluate whether the pituitary-adrenal and catecholamine response to surgical stress is modified in chronically depressed patients. DESIGN: Prospective, controlled study. PATIENTS: 25 ASA physical status I and II depressed patients taking chronic antidepressant therapy and 25 control patients undergoing orthopedic surgery of the extremities. INTERVENTIONS: All patients received anesthesia induction with thiopental 5 mg/kg and suxamethonium 1 mg/kg intravenously (IV) and were maintained with 1.5% to 2.0% isoflurane in nitrous oxide (70%) and oxygen (30%). MEASUREMENTS AND MAIN RESULTS: Plasma cortisol concentration (27.7 +/- 3.6 microg/dL) in chronic depressed patients at 60 minutes after the skin incision was not significantly higher than that (23.9 +/- 2.7 microg/dL) before the induction, although plasma cortisol concentration in the control group significantly increased. Plasma norepinephrine concentration at 60 min after the skin incision in depressed patients with more symptoms of depression was significantly higher than that of patients with less symptoms of depression. CONCLUSION: The cortisol response to surgical stress in depressed patients was inhibited and norepinephrine response to surgical stress was increased in depressed patients with more symptoms of depression. PMID- 11025240 TI - Patient perception of monetary value to avoiding unpleasant side effects of anesthesia and surgery. AB - STUDY OBJECTIVES: To determine which monetary value patients place on decreasing or eliminating common side effects of anesthesia and surgery. DESIGN: Prospective questionnaire with visual analog scales. SETTING: Preoperative holding area of a tertiary care hospital. PATIENTS: 60 adult patients scheduled for general anesthesia. INTERVENTIONS: None. MEASUREMENTS: Patients' perception of problems with previous general anesthetics and surgery and their concerns about the approaching surgery and anesthesia were recorded. Amounts (in US Dollars) that patients would be willing to pay to decrease by half or to eliminate nausea, emesis, sore throat, headache, and pain were also noted. RESULTS: 95% of the patients had previously undergone a general anesthetic, most without any problems. However, 47% of patients reported at least one problem with a previous general anesthetic, 33% had one or more concerns with the upcoming general anesthetic, and 32% would be willing to pay to decrease or eliminate one or more of these side effects. Patients who were willing to pay were more likely to have had problems or have concerns about side effects. CONCLUSIONS: Some patients would be willing to pay extra to avoid unpleasant side effects. This information can be used in quality and satisfaction surveys to justify to hospitals and payors the use of higher quality but more expensive anesthetic drugs. PMID- 11025241 TI - Sevoflurane-N2O versus propofol/isoflurane-N2O during elective surgery using the laryngeal mask airway in adults. AB - STUDY OBJECTIVES: To compare a sevoflurane-nitrous oxide (N2O) general anesthetic technique with a standard technique of propofol for induction, and isoflurane-N2O for maintenance. DESIGN: Prospective, randomized study. SETTING: University affiliated tertiary-care hospital. PATIENTS: 62 adults undergoing elective surgery using the laryngeal mask airway (LMA). INTERVENTIONS: Patients received either the standard technique of propofol for induction and isoflurane-N2O for maintenance (controls) or sevoflurane-N2O for both induction and maintenance of general anesthesia. MEASUREMENTS: Induction and emergence times, heart rate, blood pressure, oxygen saturation, and end-tidal carbon dioxide were recorded. MAIN RESULTS: Time to loss of consciousness was faster after propofol (mean +/- SEM: 51 +/- 3 sec) than after sevoflurane-N2O (85 +/- 10 sec; p < 0.05). Ready for surgery times, were however, similar between groups (10 +/- 1 vs. 11 +/- 1 min, respectively). All patients in the control group had apnea after LMA insertion compared with 4 patients in the sevoflurane-N2O group (p < 0.05). Heart rate was lower 5 and 10 minutes after LMA insertion in the sevoflurane-N2O group (69 +/- 3 and 66 +/- 3 bpm) versus the control group (81 +/- 3 bpm and 74 +/- 3 bpm, p < 0.05). After cessation of anesthetic gases, there were no differences in time to LMA removal, eye opening, or exiting the operating room (OR) between the control group (7, 8, and 10 min) and sevoflurane-N2O groups (7, 8, and 12 min, respectively). The majority of patients in both groups (92% to 97%) rated their anesthetic experience as excellent or good. CONCLUSIONS: Sevoflurane-N2O and propofol provided comparable conditions for LMA insertion. Sevoflurane-N2O was not associated with a faster return of consciousness or faster time to exit the OR compared with isoflurane-N2O. PMID- 11025242 TI - Emergence characteristics of sevoflurane compared to halothane in pediatric patients undergoing bilateral pressure equalization tube insertion. AB - STUDY OBJECTIVE: To compare the emergence characteristics of sevoflurane with halothane in pediatric patients having bilateral myringotomy and pressure equalization tube insertion using a highly standardized and common anesthetic technique. DESIGN: Prospective, randomized, double-blind study. SETTING: University hospital. PATIENTS: 43 ASA physical status I and II healthy pediatric outpatients scheduled for initial ear tube placement by one surgeon. INTERVENTIONS: Patients were randomized to receive either halothane or sevoflurane as their sole anesthetic drug for ear tube insertion. All patients as well as the research nurse grading emergence agitation were blinded to group assignment. No premedication was administered, and all patients received a standard dose of rectal acetaminophen for postoperative pain control. MEASUREMENT AND MAIN RESULTS: The primary outcome was the percentage of patients with emergence agitation, defined as thrashing behavior requiring physical restraint for greater than three minutes. Time to discharge from the postanesthesia care unit (PACU) and from the hospital were also compared. Sevoflurane patients had a greater incidence of emergence delirium: 57% versus 27% [95% CI (1.7%-58.1%) p = 0.047]. Time to discharge from the PACU and the hospital were longer for patients who received sevoflurane: 62 versus 50 min [95% CI (1.4-23.6) p = 0.02] and 102 versus 79 mins [95% CI (9.6-37.1) p = 0.003]. CONCLUSION: In our institution there is a trend toward greater emergence agitation in ear tube placement patients given sevoflurane versus halothane anesthesia. Further, (with our discharge criteria) patients are actually discharged from the PACU and the hospital faster when given halothane as a sole anesthetic when compared to sevoflurane. PMID- 11025244 TI - Noninvasive ventilation using bilevel positive airway pressure to treat impending respiratory failure in the postanesthesia care unit. AB - The author presents the use of noninvasive positive pressure ventilation using bilevel positive airway pressure (BiPAP) to treat respiratory insufficiency and hypoxemia in three patients in the postanesthesia care unit. BiPAP improved respiratory function with a decreased respiratory rate, decreased PaCO2, and improved oxygenation in the three patients. The applications of these techniques and equipment required are reviewed. PMID- 11025243 TI - Tropisetron in the prevention of postoperative nausea and vomiting. AB - STUDY OBJECTIVES: To evaluate the efficacy of tropisetron, a selective 5-HT(3) receptor antagonist, in preventing nausea and vomiting in high-risk inpatients undergoing various surgical procedures. DESIGN: Prospective, open, nonrandomized, observational, interventional study. SETTING: Postanesthesia care unit, and surgical wards of the University Hospital Center, Charleroi. PATIENTS: A total of 1,132 elective surgical inpatients (>15 years of age) in two separate surveys. The first prospective survey covered all surgical adult inpatients (n = 671) after various surgical procedures over a 3-month period. A new 3-month survey was performed to assess the effectiveness of the preventive measure and included another 461 patients. INTERVENTIONS: Risk factors associated with nausea and vomiting were recorded in the first survey and used to establish an antiemetic policy. This consisted in the administration of tropisetron 2 mg intravenously after anesthesia induction, if two patient-related risk factors associated with high-risk surgery and general anesthesia were present. MEASUREMENTS AND MAIN RESULTS: Nausea frequency and intensity, assessed every 4 hours using a visual analog scale (VAS), frequency and times of vomiting episodes and the need for rescue medication were recorded for 72 hours postoperatively. Nausea was experienced by 18.8% and vomiting by 9.8% of the patients in the first survey (211 high risk-patients of 671). In the second survey, 137 patients of 461, considered at high-risk received prophylactic tropisetron. The proportion of patients having nausea decreased to 11.1% (p,178 0.01) and vomiting episodes to 2.8% (p < 0.001). Twenty-six of the tropisetron-treated patients (19%) suffered subsequent postoperative nausea and vomiting (PONV). Patient satisfaction with tropisetron was high. CONCLUSION: Prophylactic tropisetron can reduce the incidence of PONV in selected high-risk inpatients undergoing various types of surgical procedures. PMID- 11025245 TI - Unintentional transfixation of a transverse cervical vein during placement of an interscalene brachial plexus catheter. AB - Cervical hematoma during placement of a catheter into the interscalene groove is a known complication of the procedure. I describe the occurrence of a hematoma while using a new system that is compatible with a nerve stimulator and offers the ability to aspirate during both needle placement and catheter insertion. PMID- 11025246 TI - New trends in the treatment of Hodgkin's disease. AB - Over the past several years, much progress has been made in the treatment of Hodgkin's disease (HD), with 5-year overall survival rates ranging from 60 to 95%, according to initial prognostic factors. This optimism has been tempered by the development of late complications in long-term HD survivors, in particular second neoplasms and cardiac toxicity. At present, new treatments aimed at maintaining high event-free survival and reducing long-term adverse effects are being studied. To be successful, the treatment must be tailored according to prognostic factors. PMID- 11025247 TI - Breakthroughs in the genetics of hereditary fevers. PMID- 11025248 TI - Diabetes mellitus and cancer. AB - Although an association between diabetes and cancer was found over 100 years ago, the issue underwent different interpretations over the subsequent decades, and only modern, prospective, epidemiological cohort and case-control studies conducted in several countries have provided reliable evidence of an increased cancer risk in diabetic patients, mainly in those with type 2 diabetes. This risk varies according to the tumor site: it is the greatest for primary liver cancer, moderately elevated for pancreatic cancer, and relatively low for colorectal, endometrial, breast, and renal cancers. The cause of the association is not clear and remains the subject of different hypotheses. The most frequently cited reason is the potential effect of insulin. Found in high concentrations, due to insulin resistance in most patients with type 2 diabetes, this hormone is believed to express a mitogenic effect. This hypothesis needs to be confirmed in appropriately programmed prospective studies, but it may already be helpful in choosing an adequate treatment for type 2 diabetes to achieve optimal metabolic control with a simultaneous reduction in hyperinsulinemia, such as diet, physical exercise, metformin, and acarbose. PMID- 11025249 TI - Dynamics of serum interleukin-6 level in patients with acute myocardial infarction. AB - Background: Interleukin-6 (IL-6) is a key cytokine in the initiation of the acute phase reaction that accompanies myocardial infarction. The study was designed to evaluate changes in serum IL-6 in patients with myocardial infarction and to compare IL-6 alterations with serum creatine kinase (CK) activity. Methods: Serum IL-6 level and CK activity were measured in 19 males with acute myocardial infarction. Blood was taken on the 1st, 3rd, 5th, 7th and 21st days of the disease. Control values were obtained from 30 age-matched healthy males. IL-6 was determined according to the ELISA method and CK was measured with a routine procedure. Results: Serum IL-6 was found to increase on the 1st and 3rd days of the disease, followed by some decline on days 5-21. However, the last values measured were still higher than those in the controls. Patients with transmural myocardial infarction had higher IL-6 levels than those with non-Q myocardial infarction. There was no difference in serum IL-6 in patients with myocardial infarction of the inferior cardiac wall and of the anterior cardiac wall. A correlation was found between IL-6 level and CK activity in the patients, especially on the 1st and 3rd days of the disease. Conclusion: Serum IL-6 increases in patients with myocardial infarction, and this elevation seems to be related to the mass of the affected myocardium. The highest increase is found during the first days of the disease, although the enhanced IL-6 level lasts for at least 3 weeks, probably reflecting the healing process of the myocardium. The clinical value of IL-6 determination does not seem to exceed that of the indices commonly used. PMID- 11025250 TI - Atherogen lipid profile in HIV-1-infected patients with lipodystrophy syndrome. AB - Background: Cases of lipodystrophy syndrome and metabolic disorders have been described since the onset of highly active antiretroviral therapy in HIV-infected patients. The aim of our study was to estimate the prevalence of lipodystrophy (LD) and to define the associated lipid profile of these patients. Methods: The following were determined for each patient: lipid profile (cholesterol and its subfractions, atherogenicity ratios, and triglycerides), blood glucose, and immunovirological markers (CD4(+) cell count and plasma viral load). Patients were classified into two groups on the basis of whether or not they presented with clinical signs of LD. Results: Among 233 HIV-infected patients included in the study, 61 cases (26.1%) of lipodystrophy (LD) were noted. Compared with non LD patients (NLD), LD patients were older men (P<10(-4)) with a lower CD4(+) lymphocyte cell count (P<0.007) and more often at the AIDS stage (P<10(-3)) (OR=3.2 (95% CI: 1.47-6.2)). Multivariate analysis showed a correlation between LD cases and age (10 years older) (OR=1.78 (95% CI: 1.23-2.57), P<0.002) and the decrease in CD4(+) cell count (100 CD4(+)/mm(3) lower) (OR=1.31 (95% CI: 1.09 1.58), P<0.004). An analysis of lipid subfractions and atherogenicity ratios clearly indicated a proatherogenic lipid profile for the LD patients. Conclusions: The underlying physiopathological mechanism of LD is still unknown. However, the lipid profile of HIV-1-infected patients with a LD syndrome appears to place these patients at an increased risk of progression of atherosclerosis. PMID- 11025251 TI - High-density lipoprotein cholesterol esterification and transfer rates to lighter density lipoproteins mediated by cholesteryl ester transfer protein in the fasting and postprandial periods are not altered in type 1 diabetes mellitus. AB - Background: Diabetes mellitus is associated with atherosclerosis that has, in part, been ascribed to abnormalities in the reverse cholesterol transport system. Methods: We determined, in the fasting and post-alimentary periods, rates of HDL cholesterol esterification and transfer to apoB-containing lipoproteins, cholesteryl ester transfer protein (CETP) concentration, and apoB lipoprotein size in 10 type 1 diabetics and 10 well-matched controls. Autologous HDL was labeled with [14C]cholesterol and incubated at 37 degrees C during a period of 30 min for measurement of the cholesterol esterification rate (CER), as well as for 24 h for measurement of the endogenous HDL [14C]cholesteryl ester ([14C]CE) transfer rate to apoB-containing lipoproteins after 2- and 4-h incubations with the subject's own plasma. Exogenous cholesteryl ester transfer activity (CETA) was estimated by incubation of the participant's plasma (CETP source) with [14C]CE-HDL and VLDL from a pool of plasma donors. ApoB lipoprotein size was determined using non-denaturing polyacrylamide gradient gel electrophoresis of whole plasma. Results: Contrary to previous studies, we showed that even not well controlled type 1 diabetics did not differ from lipid-matched, non-diabetic subjects in HDL-[14C]cholesterol esterification rate, transfer rates, or CETP concentration. CETP concentration correlates with the exogenous method of [14C]CE transfer and with the endogenous method only when the latter is corrected for plasma triacylglycerol (TG) concentration. In addition, during the postprandial phase, diabetic patients' VLDL are smaller and IDL size increases less than in controls. Conclusion: In type 1 diabetes mellitus, CETA is not altered when the plasma levels of donor and/or acceptor lipoproteins are within the normal range. PMID- 11025252 TI - INFLAMET: an image analyzer to display erythrocyte adhesiveness/aggregation. AB - Background: Due to a lack of simple, inexpensive, and real-time methodology, the state of erythrocyte adhesiveness/aggregation is not measured in daily practice by most clinicians. Methods: We measured the state of erythrocyte adhesiveness/aggregation before and following the administration of intravenous high-dose gamma globulins in 18 patients with various autoimmune diseases using a simple slide test and image analysis system (INFLAMET). Results: A significant (P=0.01) increment in the state of erythrocyte adhesiveness/aggregation (from 2+/ 3 to 8+/-9 arbitrary units) was noted in the entire study group despite a significant (P=0.008) decrement (from 350+/-97 to 323+/-127 mg%) in the concentration of fibrinogen and no significant change in the erythrocyte sedimentation rate (32+/-30 mm/h before and 35+/-29 mm/h following administration of gamma globulin). Conclusions: By using a simple slide test and image analysis, we were able to provide relevant rheological information that was not available from either of the routine indirect tests, i.e. the Westergren erythrocyte sedimentation rate and the concentration of fibrinogen. PMID- 11025253 TI - Histopathological and immunohistochemical study of oral lichen planus-associated HCV infection. AB - Background: In recent years, it has been suggested that oral lichen planus (OLP), a chronic inflammatory keratotic lesion, is related to hepatitis C virus (HCV) infection. Therefore, we evaluated whether the presence or absence of HCV infection caused any histopathological differences in OLP tissues. Methods; The subjects consisted of 31 patients with HCV-related liver disease complicated by OLP (32 OLP lesions) and ten OLP patients without complications due to either HCV infection or liver disease (control). A histopathological evaluation was performed in these patients. In addition, immunostaining was done on nine OLP tissues infected with HCV and on six OLP tissues without HCV infection in order to evaluate lymphocyte subsets (T cells or B cells) infiltrating into topical regions with OLP. Furthermore, the severity of hepatic fibrosis and inflammation was evaluated in liver tissues obtained by liver biopsy from six patients with HCV-related liver disease to evaluate whether there were any relationships between the severity of hepatic fibrosis or inflammation and OLP tissues. Results: There were no significant differences in the histopathological characteristics specific to OLP or in the ratios of T and B cells among infiltrating lymphocytes regardless of the presence or absence of HCV infection. Moreover, there were no certain relationships between the severity of hepatic fibrosis or inflammation and the severity of lymphocytic infiltration in OLP. Conclusions: HCV infection does not appear to influence the histopathological and immunohistochemical features of OLP. PMID- 11025254 TI - Primary leiomyosarcoma of the diaphragm in a 23-year-old male: a case report. AB - Primary neoplasms of the diaphragm are extremely rare and their diagnosis is often difficult. We present a case of leiomyosarcoma of the diaphragm in a 23 year-old male presenting with aspecific abdominal discomfort. The final diagnosis was achieved on the basis of histopathological findings after surgery. The role of different imaging techniques as diagnostic tools is also discussed. PMID- 11025255 TI - Serous retinal detachment as an early presentation of thrombotic thrombocytopenic purpura. AB - A 23-year-old woman presented with abdominal pain and blurred vision in both eyes. Fundoscopy revealed large bullous detachments of her neurosensory retina with shifting subretinal fluids bilaterally. Laboratory studies showed a hemoglobin of 6.5 mmol/l, thrombocytes 20x10(9)/l, and LDH 1098 U/l. A peripheral blood smear revealed red blood cell fragmentation. A diagnosis of thrombotic thrombocytopenic purpura was made. The patient was treated with four sessions of plasmapheresis followed by plasma infusions, which resulted in a complete recovery. PMID- 11025256 TI - Internal medicine in Italy. PMID- 11025257 TI - Polarity effects of ionization chambers used in tbi dosimetry due to cable irradiation. AB - This paper presents the results of an investigation on polarity effects in total body irradiation (TBI) dosimetry. Thimble (NE2571, 0.6 cc) and plane-parallel (Markus NE2534 0.055 cc) chambers were investigated in a 30 x 30 x 30-cm3 acrylic phantom in TBI conditions (6-MV x-rays). The thimble chamber was positioned at the midline and at the entrance and exit Dmax (1.5 cm from the phantom surface) levels. The Markus chamber, which is generally used for skin dose estimations, was positioned at various depths from the entrance surface of the phantom (from 0 to 2-cm depth). The polarity factor (Ppol) was defined as (Q+ + Q-)/2Q-, where Q+ and Q- were the collected charges at positive and negative bias voltage, respectively. The variations of Ppol with many parameters (absorbed dose, dose rate, the presence or absence of a 1-cm acrylic spoiler, irradiated cable length) were investigated. Results show that Ppol is quite small (within 1.002 for on axis measurements and 1.005 for off-axis measurements) for the NE2571 chamber when the beam spoiler is placed. Ppol was significantly higher without the beam spoiler (within 1.008 for on-axis measurements, up to 1.02 for off-axis measurements). Concerning the Markus chamber, for on-axis skin dose measurements, Ppol was found to be less than unity (around 0.988) or more than unity (around 1.0035), respectively, with and without the beam spoiler. Possible "directional effects" of the currents generated in the cable were investigated for both chambers and found to be insignificant. This shows that the application of Ppol correction has to be considered a reliable procedure in minimizing these effects. When the beam spoiler is placed, the cable has to be drawn to minimize the portion of cable just outside the beam; if this is not the case, Ppol may significantly vary (for the NE2571 chamber values up to 1.0035 were found for on axis measurements). PMID- 11025258 TI - Beam spoilers versus bolus for 6 MV photon treatment of head and neck cancers. AB - When parallel opposed 6-MV x-ray beams are used for treatment of head and neck tumors, superficial tissues and lymphatics at shallow depths of < or =4 to 6 mm may be at cancer risk but receive less than full radiation dose. In these cases, the use of either a beam spoiler or bolus material can increase dose to superficial tissues. The potential benefit of a beam spoiler relative to bolus is preservation of skin-sparing characteristics for cases in which the skin surface does not require full dose. In this study, we evaluate the application of a beam spoiler and compare it to bolus for standard treatments of head and neck tumors. Measurements of both depth dose in-water and in-air profiles were made with a beam spoiler for a 6-MV photon beam. The measurements were combined with Monte Carlo calculations to obtain the energy spectrum of the spoiler-generated electrons. An in-house pencil beam treatment-planning algorithm was used to calculate the dose distribution with spoiler. The dose distribution in the buildup region was then studied with and without the spoiler for a typical head and neck treatment with parallel-opposed beams. Dose distributions and partial volume dose histograms (PVDH) demonstrate the benefits provided by spoilers for the head and neck treatments and the limitations of their use. The beam spoiler is useful in treating the superficial lymphatics in the buildup region near head and neck tumors. Guidelines for use of beam spoiler versus bolus are discussed. PMID- 11025259 TI - Improvement in tangential breast planning efficiency using a knowledge-based expert system. AB - A knowledge-based expert system was developed for the purpose of improving radiotherapy planning efficiency for a standardized, tangential breast technique. Treatment parameters pertaining to 150 previously planned patients were used for correlating the midplane breast contour of a new patient with an appropriate set of tangential beam weights and wedge angles; other treatment parameters including, planning target volume and isocenter, were specified by a radiation oncologist. Treatment plans generated by the expert system approach and a traditional, dosimetric approach were compared and rated prospectively in 45 patients. In addition, planning time was measured for both approaches. A performance rating of 97% was achieved for the expert system, in which an artificial neural network was used to correlate breast contours to treatment parameters, and approximately 30 minutes per patient was saved in treatment planning time. This high performance rating validated various assumptions concerning the expert system: namely, that the resultant dose distribution was not influenced by tangential field width (within the range of 7 to 12 cm), nominal beam energy (6 MV), or wedge type (physical vs. enhanced dynamic). Hence, the knowledge base may be directly transferable to other cancer centers using the same breast technique, and suggests that a global resource of radiotherapy treatment plans as well as planning strategies, categorized by treatment site, stage, and technique, may be viable. PMID- 11025260 TI - Utilization of a commercial prostate brachytherapy planning system for 3D treatment planning of ophthalmic tumors. AB - The 3D brachytherapy transperineal ultrasound implant (B3DTUI) program by the Multimedia Medical System has been adopted for eye plaque treatment planning. The post op CT digitization option was selected to enter the actual seed coordinates present in the eye plaque. Although the B3DTUI algorithm is designated to calculate a permanent dose values, the described procedure allows determination of the dose distribution as the dose rate values. The dose computation program utilizes updated source quantities recommended by the AAPM TG-43 formalism, such as air kerma strength, dose rate constant, radial dose function, and anisortopy factor function. As a result, more accurate dose distribution in the target is obtained than by using traditional dose computation formalism. PMID- 11025261 TI - Simulation for localized prostate cancer: a comparison of urethrography techniques. AB - Forty-five patients having conventional fluoroscopic, or CT scan simulation of the prostate gland from January 1999 to June 1999 were studied. Patients were consecutively assigned (not randomized) in groups of 15 to 3 different urethrography techniques: air contrast alone (group 1), hypaque contrast alone (group 2), and xylocaine jelly and hypaque contrast (group 3). Outcome measures were pain scores, visualization of the apex (indicated by urethrogram tip), and frequency of corrections necessary on the basis of verification port films. Group 3 patients had the lowest mean pain score and required fewer lateral setup corrections at the time of portal imaging on the first day of treatment. A comparison of radiographs also revealed that group 2 and 3 patients (hypaque contrast) had better delineation of the prostatic anatomy than group 1 patients (air contrast). We found that of the 3 techniques tested, urethrography utilizing xylocaine jelly and hypaque was associated with the least amount of pain, least amount of corrective shifts, and best quality in defining the prostatic anatomy. PMID- 11025262 TI - Modern prostate brachytherapy. AB - As computer-aided margin tools become more sophisticated, physicists will be increasingly called upon to convert ultrasound prostate volumes to expanded planning target volumes (PTVs) to treat adequately extracapsular disease. The American Association of Physicists in Medicine Task Group 43 formalism and the new National Institute of Standards and Technology calibration system suitable for single low-energy seeds have been crucial in smoothly implementing changes in established seeds and in incorporating data from new manufacturers. However, the lack of consensus on treatment design and evaluation has led to an uncomfortably wide spectrum of clinical practice, only part of which can be attributed to variations inherent to any surgical procedure due to the practitioner's skill. The relative merits of implanting the prostate and margin with a modified uniform seed-loading approach to create plans with a relatively homogeneous dose distribution and a corresponding low risk of overdosing critical structures are addressed. Likewise, the advantages of performing postoperative dosimetry at the physically optimum time of greater than 2 weeks post implant are contrasted with the clinical advantages of obtaining the dosimetry as soon as possible. Proposed lower limits for quality parameters such D90 and V100 are reviewed. Measures of doses to the urethra, rectum, and neurovascular bundles are presented, along with correlations between various dosimetric parameters and other patient specific data with quality of life metrics involving urinary incontinence, rectal damage, and sexual dysfunction. PMID- 11025263 TI - Biological integral dose: an alternate method for numerical scoring of rival plans. AB - Numerical scoring of rival plans (NSRP) are usually based either on basis of dose volume histograms (DVH) or the relative values of corresponding normal tissue complication probabilities (NTCP) and tumor control probabilities (TCP). An alternative method for NSRP based on biological integral dose (BID) is being proposed, which is illustrated using a case of pituitary tumor planned to receive a dose of 50 Gy in 25 fractions over 5 weeks. BID for the various alternate plans -2-field (2F), 3-field (3F), 220 degrees arc (ARC) and 3-field static multileaf collimator (MLC) were calculated using the integration of the product of extrapolated response dose and the corresponding mass of the tissue enclosed separately for tumor and the normal brain in the entire planned target volume or a selected range of dose (approximately 90% and above of the normalized dose). Ratios of the BID for the brain versus the tumor were obtained and the plans were ranked on the basis of the least value of this ratio. In all of these plans, although the DVHs for normal brain were different, the DVHs for tumor were almost identical. However, the BID values for brain for 2F, 3F, ARC, and MLC were 22.53 Joules (J), 21.176 J, 21.991 J, and 10.608 J, respectively, and for tumor 0.561 J, 0.552 J, 0.555 J, and 0.556 J, respectively. The corresponding brain/tumor values were 40.16 (2F), 38.36 (3F), 39.62 (ARC), and 19.08 (MLC), thus ranking the plans in order of merit as MLC, 3F, ARC, and 2F. The BID for volumes encompassed by 90% and more of the normalized dose magnified the differences between the plans, with 2F being 29.99, compared to 3.82 for MLC. Rankings of rival plans could be based on the concept of BID. It requires a lesser number of uncertain variables and therefore could be used as an alternative technique in evaluation of the different plans in routine clinical practice. PMID- 11025264 TI - Dosimetric study of the narrow beams of 60Co teletherapy unit for stereotactic radiosurgery. AB - This study explores the possibility of using a telecobalt unit for radiosurgery. A dosimetric study was performed for the narrow beam of Cobalt 60 (60Co) unit with circular radiation fields in diameters of 11, 17, 20, 27, 32, 35, 40, and 44 mm. Percentage depth dose and off-axis ratio were measured with ion chamber and radiographic film. The tissue air ratio values derived from measurements agreed well with the calculated values for all cone sizes and depths, ranging from the depth of maximum ionization of 24 cm in water. A quantitative evaluation of treatment plans with 60Co and 6-MV photon beams was carried out. The penumbra of the narrow beam of 60Co was larger than that of the 6-MV beam by 1.3 mm on average. This difference in penumbra can be attributed to the large source size of 60Co units. The feasibility of using narrow-beam 60Co for stereotactic radiosurgery/radiotherapy is discussed. PMID- 11025266 TI - Mycoplasma pneumoniae in women. AB - Mycoplasma pneumoniae (primary atypical or Eatons' agent) is a common cause of walking or atypical pneumonia in young adults. It is responsible for approximately 43% of the cases of community-acquired pneumonia in patients between the ages of 17 and 44. Patients with Mycoplasma pneumonia typically present with the gradual onset of a nonproductive cough, sore throat, fever, and pulmonary infiltrates on chest x-ray. The diagnosis is based on symptomatology and on radiographic findings such as interstitial infiltrates, patchy infiltrates, plate-like atelectasis, nodular infiltration, or hilar adenopathy. The primary treatment for Mycoplasma pneumoniae is a macrolide antibiotic, preferably azithromycin: 500 mg initially and then 250 mg daily for 4 days. PMID- 11025265 TI - Measurement of relative output for 90Sr ophthalmic applicators using radiochromic film. AB - The treatment of various superficial lesions of the eye has, for many years, been conducted using strontium 90 (90Sr) ophthalmic applicators that have a steep dose gradient near their surface. A new applicator acquired by a treatment facility must have its output compared with that of any older applicators already in use to ensure consistent treatments. These measurements may be done using available dosimeters such as film and thermoluminescent detectors. Our work made use of radiochromic film and a document scanner to perform relative output measurements for 4 different 90Sr ophthalmic applicators acquired from the same manufacturer (Amersham Healthcare, Arlington Heights, IL) over a span of 28 years. Relative outputs were found to vary by < 10% with respect to the manufacturer's values, which is well within the uncertainty limit for absolute output of 20% specified by the manufacturer. The film measurements were verified using thermoluminescent dosimeters. Radiochromic film was also used to obtain a percentage depth dose curve and a 2 dimensional isodose distribution in a plane perpendicular to the active surface for the newest applicator (SIA 20). PMID- 11025267 TI - Strategic planning: a modern tool for obstetricians and gynecologists in a competitive practice environment. AB - The now inevitable changes in health care practice and financing have resulted in missed opportunities for physician directed improvements in medical and surgical care of patients. The process of strategic planning is the business tool that allows individuals and organizations to assess proactively the need for change and allows them to adapt and respond appropriately to a changing environment. The fundamental components of implementing a strategic plan include assessing environmental and organizational factors, defining strengths and limitations of the organization, identifying a market niche if possible, and, after establishing key objective and goals, periodically reviewing the plan to determine that goals are attained. Future success in health care will likely depend upon the ability to effectively anticipate, plan, and implement necessary changes. PMID- 11025268 TI - Bacterial vaginosis. AB - Bacterial vaginosis is the most common cause of vaginitis, affecting over 3 million women in the United States annually. Depopulation of lactobacilli from the normal vaginal flora and overgrowth of Gardnerella vaginalis and other anaerobic species are the presumed etiology. To date, no scientific evidence shows that bacterial vaginosis is a sexually transmitted disease. Malodorous vaginal discharge is the most common symptom. Differential diagnoses include trichomoniasis, moniliasis, and allergic or chemical dermatitis. The diagnosis is confirmed when at least three of the following four findings are present (Amsel's criteria): 1) thin, homogenous discharge, 2) pH greater than 4.5, 3) positive amine test, and 4) presence of clue cells. The sensitivity and positive predictive value are both 90%. Vaginal Gram stain is also reliable and allows for permanent record. Cultures are nonspecific because G. vaginalis resides in normal vaginal flora as well. Papanicolaou smears are not particularly sensitive, but their positive predictive value is very high. The Centers for Disease Control and Prevention recommend three treatment regimens in nonpregnant patients: oral metronidazole (500 mg twice daily for 7 days), intravaginal 2% clindamycin cream (one applicatorful at bedtime for 7 days), or intravaginal metronidazole gel (one to two applicatorfuls per day for 5 days). Alternative regimens include a single 2-g oral dose of metronidazole or a 7-day course of oral clindamycin, 300 mg twice daily. The association between bacterial vaginosis and adverse pregnancy outcomes has satisfied many criteria for a causal inference. Treatment of bacterial vaginosis in women with previous history of preterm labor results in fewer preterm deliveries than in untreated women from the same population. PMID- 11025269 TI - Syphilis in women. AB - Infection with the spirochete Treponema pallidum causes syphilis. Transmission of syphilis occurs through sexual contact with persons who have infectious mucocutaneous lesions. Before the advent of penicillin, this infection was responsible for a large portion of the debilitated patients residing in mental institutions. Later during this century, the disease became somewhat uncommon, only to see a resurgence during the early part of this decade. Syphilis progresses through stages defined as primary, secondary, latent, and tertiary. Diagnosis of this infection is by clinical examination, dark-field microscopy, and serology. Penicillin is the treatment of choice for all stages of syphilis. Doxycycline and tetracycline are acceptable alternatives in some penicillin allergic patients. If patients are pregnant or have central nervous system involvement, alternative regimens should not be used because of lack of efficacy and/or fetal toxicity. Therefore, these patients must be desensitized and treated with penicillin. No effective vaccine for this infection is currently available. PMID- 11025270 TI - When your patient needs mental health care. AB - Whether as primary care, specialty, or subspecialty physicians, obstetrician/gynecologists encounter many patients with mental health problems. Although fortunately, the stigma of mental illnesses and treatments has somewhat decreased and the accuracy of diagnosis and efficacy of care have increased, it has unfortunately become more complicated for patients to access that care. There are more and more mental health practitioners and disciplines, making it difficult for the referring ob/gyn and the patient to identify which sort of professional they need. There is an increasing array of third-party payer arrangements, each of which has constantly changing rules and procedures. This article illustrates some typical cases and summarizes some commonsense approaches. Its purpose is to help the busy practitioner connect patients with the care they need. PMID- 11025271 TI - The onset of human labor: current theories. AB - Innovative research has led to marked improvement in medical knowledge. Advances in technology have allowed us to attain a clearer understanding of many physiologic processes. Despite these impressive gains in knowledge, the cause of the initiation of the process of human labor remains unclear. Potential factors include changes in hormonal levels of estrogen and progesterone, increased production of prostaglandins, and elevation of levels of corticotropin-releasing hormone, as well as increased sensitivity of the myometrium to endogenous oxytocin. It is most likely that interactions between these factors play an essential role in the process of labor, but the precise mechanism of onset still eludes us. PMID- 11025272 TI - Infections and infertility. AB - Infertility affects 10-15% of all couples. Pelvic infections are an important cause of infertility, primarily as a result of tubal damage. Damage to the fallopian tubes from infections may be due to adhesions, tubal mucosal damage, or tubal occlusion that interferes with normal ovum transport. The infections most commonly related to infertility include gonorrhea, chlamydia, and pelvic inflammatory disease. Tuberculosis also is a common cause of infertility in Third World nations. Sequelae resulting from these infections include ectopic pregnancy, infertility, chronic pelvic pain, hydrosalpinx, and tuboovarian abscess. Neisseria gonorrhoeae and Chlamydia trachomatis are the primary causes of pelvic inflammatory disease. Chlamydial infections may be asymptomatic, and the resulting salpingitis is often referred to as silent pelvic inflammatory disease. Polymicrobial infection with other organisms such as anaerobes or facultative aerobes may be initiated by gonorrhea, chlamydia, or both. Early recognition of infection, prompt institution of appropriate antibiotic therapy, and proper follow-up are important to prevent the sequelae of pelvic inflammatory disease. Surgical intervention may be needed to treat immediate or long-term sequelae of infection. Prevention of pelvic infections should be a high priority. Fortunately, treatment options such as tubal microsurgery and assisted reproductive technologies offer couples reproductive options even when infertility occurs as the result of a previous pelvic infection. PMID- 11025273 TI - Smoke alarm: a review of the clinical impact of smoking on women. AB - This article is a review of the health impacts of smoking on women and their newborns, with an overview of the prevalence of smoking, mortality statistics, and costs. Over 50 disorders related to smoking are identified. These include many chronic and severe problems that are often overshadowed by the more well known impacts of cancer and cardiac disease. Female impacts of smoking include two-fold rates of ectopic pregnancy, premature delivery, and low-birthweight infants; earlier menopause; pelvic-floor disorders, including urinary incontinence; menstrual disturbances; decreased fertility; greater risk of oral contraceptive failure; and babies with increased rates of sudden infant death syndrome, asthma, and attention-deficit disorders. From a clinical perspective, one in five women is addicted to nicotine. In addition, women smokers experience a stronger response to nicotine and incur higher rates of lung cancer and cardiovascular disease than their male counterparts. PMID- 11025275 TI - Error correction and the basal ganglia. Response to Lawrence (2000). PMID- 11025274 TI - Error correction and the basal ganglia: similar computations for action, cognition and emotion? PMID- 11025276 TI - Go with the flow. PMID- 11025277 TI - Sharing - a new culture? PMID- 11025278 TI - Gaze into my eyesellipsis. PMID- 11025279 TI - Locating the language switch. PMID- 11025280 TI - Consciousness: mapping the theoretical landscape. AB - What makes us conscious? Many theories that attempt to answer this question have appeared recently in the context of widespread interest about consciousness in the cognitive neurosciences. Most of these proposals are formulated in terms of the information processing conducted by the brain. In this overview, we survey and contrast these models. We first delineate several notions of consciousness, addressing what it is that the various models are attempting to explain. Next, we describe a conceptual landscape that addresses how the theories attempt to explain consciousness. We then situate each of several representative models in this landscape and indicate which aspect of consciousness they try to explain. We conclude that the search for the neural correlates of consciousness should be usefully complemented by a search for the computational correlates of consciousness. PMID- 11025281 TI - Interdimensional interference in the Stroop effect: uncovering the cognitive and neural anatomy of attention. AB - In the classic Stroop effect, naming the color of an incompatible color word (e.g. the word RED printed in green ink; say, 'green') is much slower and more error-prone than is naming the color of a control item (e.g. XXX or CAT printed in green; say 'green'). This seemingly simple interference phenomenon has long provided a fertile testing ground for theories of the cognitive and neural components of selective attention. We present a sketch of the behavioral phenomenon, focusing on the idea that the relative automaticity of the two dimensions determines the direction and the degree of interdimensional interference between them. We then present an outline of current parallel processing explanations that instantiate this automaticity account, and we show how existing interference data are captured by such models. We also consider how Stroop facilitation (faster response of 'red' to RED printed in red) can be understood. Along the way, we describe research on two tasks that have emerged from the basic Stroop phenomenon - negative priming and the emotional Stroop task. Finally, we provide a survey of brain imaging research, highlighting the possible roles of the anterior cingulate in maintaining attentional set and in processing conflict or competition situations. PMID- 11025282 TI - Escape from illusion: reducing false memories. AB - Illusory memories are unsettling, but far from uncommon. Over the past several years, increasing experimental and theoretical attention has focused on misattribution errors that occur when some form of memory is present but attributed to an incorrect time, place or source. Demonstrations of errors and distortions in remembering raise a question with important theoretical and practical implications: how can memory misattributions be reduced or avoided? We consider evidence that documents the occurrence of illusory memories, particularly false recognition responses, and then review three ways in which memory distortion can be minimized. PMID- 11025283 TI - Individual differences in music performance. AB - Music cognition depends on the existence and deployment of processes for detecting, storing and organizing musical materials according to underlying structural features. Common cultural experiences develop these processes to a certain degree, but specifically designed and supported learning environments are required to achieve the levels of expertise required to perform western art music. Certain motivational and social factors are therefore implicated in the maintenance of activities that promote skill-acquisition, such as practice. Expert musical performance is not just a matter of technical motor skill, it also requires the ability to generate expressively different performances of the same piece of music according to the nature of intended structural and emotional communication. This review examines these abilities and describes how some of them have been shown to have lawful relationships to objective musical and extra musical parameters. Psychological research is thus engaged in a process of demystifying musical expertise, a process that helps to improve upon culturally prevalent, but ultimately non-explanatory, notions of inborn 'talent'. PMID- 11025284 TI - Erratum. PMID- 11025285 TI - Na3Ca2TaO6, a rock-salt superstructure phase with a fully ordered cation arrangement AB - The title quaternary oxide, trisodium dicalcium tantalum hexaoxide, is isostructural with Li(3)Ni(2)TaO(6), a partially ordered rock-salt phase. The Na, Ca and Ta atoms occupy octahedral sites in an orderly manner and form a cation ordered superstructure. PMID- 11025286 TI - On the way to sol-gels: an analysis of the intra- and intermolecular hydrogen bonding in AB - We have been able, via a new synthetic route, to obtain a complete crystal structure of the title compound, tetraaquabarium hydroxide iodide, [Ba(OH)I(H(2)O)(4)], for which the heavy atoms only were characterized by Kellersohn, Beckenkamp & Lutz [Z. Naturforsch. Teil B (1991), 46, 1279-1286]. In the present results, the H-atom positions could be located using X-ray data collection at low temperature. A three-dimensional network is built up via hydrogen bonds. It was also observed that the title compound undergoes hydrolysis and might therefore be regarded as an intermediate in the formation of sol-gels, starting from BaI(2) and leading to [Ba(OH)(2)(H(2)O)(x)]. PMID- 11025287 TI - Potassium sodium tin selenide, K3NaSn3Se8 AB - The title compound, tripotassium sodium tritin octaselenide, K(3)NaSn(3)Se(8), has a molecular (zero-dimensional) structure containing trimeric [Sn(3)Se(8)](4-) units which consist of three edge-sharing SnSe(4) tetrahedra. The [Sn(3)Se(8)](4 ) anions and the tetrahedrally coordinated Na(+) cations are arranged in an alternating fashion along the c axis to form SiS(2)-like chains, which are then separated by eight-coordinate K(+) cations. The Sn-Se bond distances are normal, being in the range 2.477 (1)-2.612 (1) A. PMID- 11025288 TI - Tetra-n-butylammonium bis(1,2-dimercaptoethene-1, 2-dicarbonitrilato S,S')nickel(III) AB - The crystal structure of the title compound, (C(16)H(36)N)[Ni(C(4)N(2)S(2))(2)], shows stacking of the dimerized anions, surrounded by columns of cations. PMID- 11025289 TI - A novel PtII-dibenzo-18-crown-6 (DB18C6) complex AB - The novel Pt(II)-dibenzo-18-crown-6 (DB18C6) title complex, &mgr; [tetrakis(thiocyanato-S)platinum(II)]-N:N'-bis[2,5,8,15,18, 21 hexaoxatricyclo[20.4.0.1(9,14)]hexacosa-1(22),9(14),10,12,23, 25-hexaene kappa(6)O]potassium(I), [K(C(20)H(24)O(6))](2)[Pt(SCN)(4)], has been isolated and characterized by X-ray diffraction analysis. The structure analysis shows that the complex displays a quasi-one-dimensional infinite chain of two [K(DB18C6)](+) complex cations and a [Pt(SCN)(4)](2-) anion, bridged by K(+).pi interactions between adjacent [K(DB18C6)](+) units. PMID- 11025290 TI - (&mgr;-2, 6-Bis AB - The binuclear cation of the title compound, [Ni(2)(C(33)H(29)N(4)O(3))(H(2)O)(4)]C(2)H(3)O(2).C(3)H(7)NO.0. 75H(2)O, was synthesized as a model for the active site of urease. Two tridentate halves of the symmetrical 2, 6-bis[(2-hydroxyphenyl)(2-pyridylmethyl)amino]methyl-4 methylphenol ate (BPPMP(3-)) ligand are arranged in a meridional fashion around the two Ni(II) ions, with the phenoxo O atom bridging the Ni(II) ions. The cation has an approximate twofold rotation axis running through the C-O bond of the bridging phenolate group. Four water molecules complete the octahedral environment of each Ni(II) ion. PMID- 11025291 TI - trans-Dichloro(1,12-diphenyl-5,8-dioxa-2,11-dithiadodecane-S, S')palladium(II) AB - In the title compound, [PdCl(2)(C(20)H(26)O(2)S(2))], the Pd atom has a distorted square-planar coordination geometry, with Pd-S distances of 2.3121 (18) and 2.3102 (18) A, Pd-Cl distances of 2.291 (2) and 2.314 (2) A, and S-Pd-Cl angles of 94.98 (7) and 86.25 (7) degrees. Upon complexation, an 11-membered ring is formed by the S-Pd-S linkage in the ligand chain. PMID- 11025292 TI - Bis(dimethyl sulfoxide-S)tetrakis(&mgr;-p-hydroxybenzoato-O:O')dirhodium(II) tetra kis(&mgr;-butyrato-O:O')bis(dimethyl sulfoxide-S)dirhodium(II) cocrystal ethanol disolvate AB - The title structure, [Rh(2)(C(7)H(5)O(3))(4)(C(2)H(6)OS)(2)]. [Rh(2)(C(4)H(7)O(2))(4)(C(2)H(6)OS)(2)].2C(2)H(6)O, contains two discrete neutral Rh-Rh dimers cocrystallized as the ethanol disolvate. Each dimer is situated on an inversion center. The butyrate chain displays disorder in one C-atom position. In each dimer, the dimethyl sulfoxide ligand (dmso) is bound via S, as expected. The ethanol is a hydrogen-bond acceptor for one p-hydroxybenzoate hydroxyl group and acts as a hydrogen-bond donor to the dmso O atom of a neighboring p hydroxybenzoate dirhodium complex. A third hydrogen bond is formed from the other p-hydroxybenzoate hydroxyl group to the dmso O atom of a butyrate-dirhodium complex. PMID- 11025293 TI - Tetrabromo(2,6-dimethylpyridine-N)titanium(IV), a twinned crystal structure AB - The reaction of 2,6-dimethylpyridine with TiBr(4) affords the title compound, [TiBr(4)(C(7)H(9)N)], which is the first example of a neutral TiBr(4)L complex (L is a singly bonded ligand). The environment around the Ti atom can be described as a somewhat distorted trigonal bipyramid, with the nitrogen base occupying an equatorial position. The crystal was a non-merohedral twin. PMID- 11025294 TI - Two mixed-metal carboxylate-base adducts AB - The crystal structures of hexa-&mgr;-propionato-1:2kappa(6)O:O';1:3kappa(6)O:O' diquinoline-2ka ppaN,3kappaN-calcium(II)dizinc(II), [CaZn(2)(C(3)H(5)O(2))(6)(C(9)H(7)N)(2)], and hexa-&mgr;-pivalato 1:2kappa(6)O:O';1:3kappa(6)O:O'-diquinoline-2kapp aN,3kappaN calcium(II)dicobalt(II), [CaCo(2)(C(5)H(9)O(2))(6)(C(9)H(7)N)(2)], are described. Both contain a linear array of one Ca(II) ion and two M(II) (M = Zn, Co) ions connected by two sets of three carboxylate ligands in syn-syn bridging modes. The distorted tetrahedral geometry around the M(II) ion is completed by a quinoline N atom. The central Ca(II) ion occupies a crystallographic inversion centre and is octahedrally coordinated by six carboxyl O atoms in each structure. The Zn(II). Ca(II) and Co(II).Ca(II) distances are 3.8504 (9) and 3.7929 (5) A, respectively. PMID- 11025295 TI - Diaqua6,6'-dimethoxy-2,2'- AB - In the title compound, [Ni(C(19)H(20)N(2)O(4))(H(2)O)(2)], the Ni atom has a distorted octahedral coordination geometry in which the tetradentate Schiff base ligand acts as a cis-N(2)O(2) donor defining an equatorial plane, and water molecules occupy the axial positions. The two parts of the molecule are related by a mirror plane that passes through the Ni atom and is perpendicular to the equatorial plane. The angular distortions from normal octahedral geometry are in the range 1-6 degrees, and the equatorial plane, defined by the donor atoms of the Schiff base, is almost square planar. The six-membered ring comprising the Ni, the imine N and the propylene C atoms adopts a half-chair conformation. The Ni-O [2.017 (2) A] and Ni-N [2.071 (2) A] distances are within the ranges expected for high-spin octahedral nickel complexes. PMID- 11025296 TI - A ferrocenyl-substituted pseudotitanocene complex AB - The title compound, (eta(5)-cyclopentadienyl)[(1,2,3,4, 5-eta)-4-ferrocenyl 1,2,5,6-tetrakis(trimethylsilyl)cyclohexa-2, 4-dien-1-yl]titanium(II), [TiFe(C(5)H(5))(2)(C(23)H(42)Si(4))] or [Tieta(5)-C(6)H(2)Fe(eta(5) C(5)H(4))(eta(5)-C(5)H(5))Si(CH(3))(3 )(4)(eta(5)-C(5)H(5))], possesses two directly linked metallocene units that subtend an angle of 52.9 (1) degrees (defined by the least-squares planes of the directly connected pi-ligands) associated with the steric requirements of the bulky trimethylsilyl substituents. The cyclohexadienyl ligand adopts an envelope conformation; the perpendicular distance of its eta(5)-plane to the Ti atom is 1.512 (1) A. PMID- 11025297 TI - mu-Oxo-bis[(2,3,12,13-tetrabromo-5,10,15, 20-tetraphenylporphyrinato)iron(III)] bis(dichloromethane) solvate. AB - The precise structure of the title compound, [Fe(2)O(C(44)H(24)Br(4)N(4))(2)].2CH(2)Cl(2), is reported. The Fe-N distances are non-equivalent in pairs because of the asymmetric peripheral substitution; the values are 2.098 A to the brominated rings and 2.041 A to the other two rings. The Fe-O bond distance is 1.7583 (4) A. The molecule has required twofold symmetry so that there is one unique porphyrin macrocycle and one Fe-O bond length in contrast to a previous report on the same species. PMID- 11025299 TI - A novel three-dimensional copper(II) network via hydrogen bonds: diaquabis AB - The crystal structure of the title complex, [Cu(C(7)H(8)N(4))(2)(H(2)O)(2)](ClO(4))(2), consists of a discrete centrosymmetric [Cu(C(7)H(8)N(4))(2)(H(2)O)(2)](2+) cation and two perchlorate anions. The Cu(II) centre is six-coordinated by four N donors from the two pyrazole rings [Cu-N 1.998 (2) and 2.032 (3) A] and two O atoms from the water molecules occupying the apical sites [Cu-O 2.459 (3) A]. The coordination geometry of the complex can be described as octahedral. There is a unique three dimensional network in which the perchlorate units are linked by a combination of strong O-H.O and weak C-H.O hydrogen bonds. PMID- 11025298 TI - Chloramine-B sesquihydrate. AB - In the title compound, sodium N-chlorobenzenesulfonamide sesquihydrate, Na(+).C(6)H(5)ClNO(2)S(-).1.5H(2)O, the sodium ion exhibits octahedral coordination by O atoms from three water molecules and by three sulfonyl O atoms of three different N-chlorobenzenesulfonamide anions. A two-dimensional polymeric layer consists of units, each comprising two face-sharing octahedra which share four corners with four other such units, the layer running parallel to the ab plane. The water molecules participate in hydrogen bonds of the types O-H...O, O H...N and O-H...Cl. PMID- 11025300 TI - A dicationic gallium-oxo-hydroxide cage compound AB - The crystal structure of hexa-&mgr;-hydroxo-dodecamethylocta-&mgr;(3)-oxo dodecagallium(III) bis[tetrakis(pentafluorophenyl)borate(III)] chlorobenzene disolvate dihydrate, [Ga(12)(CH(3))(12)(&mgr;(3)-O)(8)(&mgr; OH)(6)](C(24)F(20)B)(2). 2C(6)H(5)Cl.2H(2)O, is reported. The gallium-oxo hydroxide dication is located on an inversion center and adopts a cage structure composed of 12 fused Ga(3)O(3) rings and is associated with the hydrate molecules and the [B(C(6)F(5))(4)](-) anions through hydrogen bonds and one O-H.pi-ring interaction. Disordered chlorobenzene solvent molecules are also present in the crystal structure. PMID- 11025301 TI - Astemizole tetrachlorocuprate(II). AB - The structure of ?3-[(4-fluorophenyl)methyl]-1H-benzimidazol-2-ylidene??1-[2-(4- methox yphenyl)ethyl]-4-piperidin-1-io?ammonium tetrachlorocuprate(II), (C(28)H(33)FN(4)O)[CuCl(4)], contains diprotonated cations of astemizole hydrogen bonded to three Cl atoms in two different CuCl(4)(2-) anions, with Cl.N distances in the range 3.166 (4)-3.203 (4) A. The geometry around copper is flattened tetrahedral with significantly different Cu-Cl distances which lie in the range 2.1968 (14)-2.2861 (12) A. The phenylethyl C atoms of the (4-methoxyphenyl)ethyl group are disordered indicating the presence of two conformers in the crystals. PMID- 11025302 TI - Hydrogen bonds and C-H.O interactions in N-(2-hydroxy-1, 1 dimethylethyl)benzamide at 150 K AB - The title compound, C(11)H(15)NO(2), crystallized in the centrosymmetric space group P2(1)/n with one molecule in the asymmetric unit. There is a single intermolecular hydrogen bond, in which the N(donor).O(acceptor) distance is 3.0374 (11) A and the N-H. O angle is 171.0 (12) degrees. The single intramolecular hydrogen bond has an O(donor).O(acceptor) distance of 2.6279 (11) A and an O-H.O angle of 161.8 (14) degrees. The four leading intermolecular C-H.O interactions have H.O distances ranging from 2.52 to 2.65 (2) A and C-H.O angles ranging from 125.2 (9) to 143 degrees. Chains of interactions form two dimensional networks. PMID- 11025303 TI - 2,4,5,7-Tetramethylphenanthrene at 150 K AB - The title compound, C(18)H(18), crystallized in the centrosymmetric space group P2(1)/c with one molecule as the asymmetric unit. The methyl-group H atoms at the 4 and 5 positions are ordered, while those at the 2 and 7 positions are disordered. The torsion of the bay region of the core is notably similar to that of other 4, 5-dimethylphenanthrenes. No substantial C-H.pi interaction occurs in this structure. PMID- 11025304 TI - Hydrogen bonding and C-H.O interactions in 2- AB - The title acid, C(19)H(16)O(2), crystallized in the centrosymmetric space group Pnna with three molecules in the asymmetric unit (apparently the first reported instance of Z' = 3 in this space group). Four intermolecular hydrogen bonds have O(D).O(A) distances of 2.660 (2), 2.594 (2), 2.633 (2) and 2.646 (2) A, and angles of 177 (2), 179 (2), 175 (2) and 175 (2) degrees. The four hydrogen bonds form two R(2)(2)(8) cyclic dimers, one about a twofold axis. Five leading intermolecular C-H.O interactions are present. Although first- and second-level graph sets involving these nine interactions are dominated by finite patterns, a three-dimensional network becomes evident upon analysis of higher-level graphs. A number of intramolecular C-H.O interactions are also present. PMID- 11025305 TI - The antifolate trimetrexate: observation of the enzyme-binding conformation. AB - The crystal structure of the title compound contains four 2, 4-diamino-5-methyl-6 [(3,4,5-trimethoxyanilino)methyl]quinazoline molecules, two dimethyl sulfoxide molecules and three water molecules in the asymmetric unit, i.e. 4C(19)H(23)N(5)O(3).-2C(2)H(6)OS.3H(2)O. All four quinazoline molecules adopt trans,-gauche conformations. An extensive hydrogen-bond network involving N. N base-pairing interactions, as well as the dimethyl sulfoxide and water molecules, stabilizes the crystal structure. PMID- 11025306 TI - Three methoxy-substituted diethyl 4-phenyl-2,6-dimethyl-1, 4-dihydropyridine-3,5 dicarboxylate compounds. AB - Diethyl 4-(2,5-dimethoxyphenyl)-2,6-dimethyl-1,4-dihydropyridine-3, 5 dicarboxylate, C(21)H(27)NO(6), (I), diethyl 4-(3, 4-dimethoxyphenyl)-2,6 dimethyl-1,4-dihydropyridine-3, 5-dicarboxylate, C(21)H(27)NO(6), (II), and diethyl 2, 6-dimethyl-4-(3,4,5-trimethoxyphenyl)-1,4-dihydropyridine-3, 5 dicarboxylate, C(22)H(29)NO(7), (III), crystallize with hydrogen-bonding networks involving the H atom bonded to the N atom of the 1,4-dihydropyridine ring and carbonyl O atoms in (I) and (II). Unusually, (III) shows O atoms of methoxy groups serving as hydrogen-bond acceptors. PMID- 11025307 TI - A synchrotron-radiation study of the lactone form of rhodamine B at 120 K AB - The structure of the lactone form of rhodamine B, 3, 6-bis(diethylamino)-1',3' dihydrospiro[xanthene-9, 3'-isobenzofuran]-1'-one, C(28)H(30)N(2)O(3), has been determined at 120 K using synchrotron radiation at a wavelength of 0.496 A. The structure contains two independent rhodamine B molecules with virtually identical geometry. The xanthene main planes of the molecules are inclined at an angle of 41.6 (2) degrees to one another. Molecule 2 has a statistically disordered ethyl group, with 71% in one orientation and 29% in a second orientation. The lactone C O bonds are 1.497 (1) and 1.495 (1) A. There are no classical hydrogen bonds, but the structure is stabilized by two short C.O interactions. The crystals of the lactone form were produced by a novel hydrothermal reaction. PMID- 11025308 TI - Bis(4-propargyloxy-3-quinolylthio)methane. AB - The molecular structure of the title compound, C(25)H(18)N(2)O(2)S(2), in the crystal is characterized by almost parallel quinoline and propargyl groups that point in opposite directions out of the quinoline planes. Intermolecular C[triple bond]C-H...N hydrogen bonding is observed, but the hydrogen-bond geometry is poor. PMID- 11025309 TI - trans-cis S-benzyl dithiocarbazate AB - In the crystal structure of the title compound, C(8)H(10)N(2)S(2), the molecules are linked by N-H.S hydrogen bonds between the imino group and the thione-S atoms to form a chain along the b axis. The dithiocarbazate moiety is rotated by 85.8 (2) degrees with respect to the phenyl ring. PMID- 11025310 TI - 4-(N-Hydroxyethyl-N-methylamino)benzaldehyde thiosemicarbazone AB - In the crystal structure of the title compound, C(11)H(16)N(4)OS, the phenyl ring and the thiosemicarbazone moiety from a dihedral angle of 7.7 (1) degrees. The crystal structure is governed by N-H.O and O-H.S hydrogen bonds leading to the formation of a two-dimensional network. PMID- 11025311 TI - 2,4-Bis(o-tolyl)-3-azabicyclo AB - The crystal structure of the title compound, C(22)H(25)NO, confirms that the bicyclic ring system adopts the chair-chair conformation. The phenyl rings are equatorially disposed with respect to the bicyclic ring. There is a slight deviation from the chair conformation in the case of the cyclohexane ring. PMID- 11025312 TI - trans-Tetrachlorobis(N, N'-dimethylimidazolidine-2-thione)tellurium(IV), a thiourea complex of tellurium with asymmetric Te-S bonds AB - The structure of the title compound, [TeCl(4)(C(5)H(10)N(2)S)(2)] or C(10)H(20)Cl(4)N(4)S(2)Te, has been solved in order to study the stereochemical activity of the lone pair of electrons on Te(IV). The two crystallographically independent molecules in the asymmetric unit both show a distorted octahedral coordination of the Te atom. The two Te-S bonds are trans to each other in both molecules and are greatly asymmetric, with bond lengths of 2.5686 (7) versus 2.8557 (8) A and 2.5859 (7) versus 2.8165 (9) A. The Te-Cl bond lengths lie in the range 2.5236 (7)-2.5589 (8) A. The asymmetric Te-S bonds and a large S-Te-Cl angle of ca 97 degrees involving the long Te-S bonds indicate stereochemical activity of the lone pair of electrons on Te. PMID- 11025313 TI - The addition-reaction product of 1,1,1,4,4,4-hexachloro-1, 4-disilabutane with N methylimidazole AB - The product of the addition reaction of 1,1,1,4,4,4-hexachloro-1, 4-disilabutane with N-methylimidazole is &mgr;-ethylene-C(1):C(2)-bis[dichlorotris(1 methylimidazole-N(3))sili con(IV)] dichloride, C(26)H(40)Cl(4)N(12)Si(2)(2+).2Cl( ). Two of the six Cl atoms are replaced by aromatic nitrogen bases and the coordination sphere of silicon is extended from four to six. The molecule is located on a crystallographic centre of inversion. The environment around the Si atom can be described as a slightly distorted octahedron with the Cl atoms occupying axial positions and the three N-methylimidazole ligands and the ethylene bridge in the equatorial plane. PMID- 11025314 TI - Multicentre hydrogen bonds in a 2:1 arylsulfonylimidazolone hydrochloride salt. AB - The title compound, (S)-(+)-4-[5-(2-oxo-4, 5-dihydroimidazol-1-ylsulfonyl)indolin 1 -ylcarbonyl ]anilinium chloride (S)-(+)-1-[1-(4-aminobenzoyl)indoline-5- sulfonyl]-4-phenyl-4, 5-dihydroimidazol-2-one, C(24)H(23)N(4)O(4)S(+).Cl(-). C(24)H(22)N(4)O(4)S, crystallizes in space group C2 from a CH(3)OH/CH(2)Cl(2) solution. In the crystal structure, there are two different conformers with their terminal C(6) aromatic rings mutually oriented at angles of 67.69 (14) and 61.16 (15) degrees. The distances of the terminal N atoms (of the two conformers) from the chloride ion are 3.110 (4) and 3.502 (4) A. There are eight distinct hydrogen bonds, i.e. four N-H...Cl, three N-H...O and one N-H...N, with one N-H group involved in a bifurcated hydrogen bond with two acceptors sharing the H atom. C H...O contacts assist in the overall hydrogen-bonding process. PMID- 11025315 TI - Two crystalline modifications of 2-hydroxycyclopent-2-enone AB - The orthorhombic form of 2-hydroxycyclopent-2-enone, C(5)H(6)O(2), consists of chains of hydrogen-bonded molecules aligned along a twofold screw axis. The monoclinic form contains two independent molecules, which have different orientations of the hydroxyl proton, and which assemble into ribbons along a twofold screw axis. PMID- 11025316 TI - Three leflunomide metabolite analogs. AB - The title compounds, 1-cyano-2-hydroxy-N-[4-(methylsulfonyl)phenyl]but-2-enamide, C(12)H(12)N(2)O(4)S, PHI492, 1-cyano-2-hydroxy-N-[3-(methylsulfonyl)phenyl]but-2 enamide, C(12)H(12)N(2)O(4)S, PHI493, and N-[3-bromo-4-(trifluoromethoxy)phenyl] 1-cyano-2-hydroxybut-2-e namide , C(12)H(8)BrF(3)N(2)O(3), PHI495, are potent inhibitors of Bruton's tyrosine kinase (BTK). The molecular structures of these compounds are similar and they display similar hydrogen-bonding networks and crystal packing. Examination of the crystal-packing interaction in the three compounds reveals an alternating direction of adjacent molecules in the crystalline lattice due to intermolecular cyano-amide hydrogen bonding. PHI492, a positional isomer of PHI493, does not form intermolecular O-H.O hydrogen bonds between molecules and crystallizes in a space group different from that of PHI493 and PHI495. The aromatic ring and the amide group of each molecule form a conjugated pi-system which ensures planarity, with further stabilization gained from intramolecular O-H.O hydrogen bonds. PMID- 11025317 TI - 2-(2-Oxoimidazolidinyl)ethyl 2-methylprop-2-enoate AB - In the crystal structure of the title compound, C(9)H(14)N(2)O(3), the molecules are linked by N-H.O=C bonds into chains parallel to [001]. Large crystals are readily obtained, presumably because of the hydrogen bonds and an energetically stable conformation of the molecule. PMID- 11025318 TI - 5-Oxocyclooctanecarboxylic acid: hydrogen-bonding pattern and conformational disorder in a medium-ring varepsilon-keto acid AB - Molecules of the title compound, C(9)H(14)O(3), adopt a chiral 'boat-chair' conformation, in which the carboxyl group avoids potential cross-ring ketone interactions by an outward 'equatorial' orientation. The asymmetric unit contains two such molecules, one conformationally fixed without disorder, (I), and the other, (I'), extensively disordered, both in the bond lengths and angles of the carboxyl and by a coupled 'up-down' conformational disordering [ratio of 60:40 (1)] of the remote ends of the boat-chair system. Each molecule in the asymmetric unit forms a centrosymmetric hydrogen-bonded carboxyl dimer with a second molecule of its own type. For (I), O.O = 2.658 (3) A and O-H.O = 174 degrees. For (I'), O.O = 2.653 (3) A and O-H.O = 165 degrees. A number of intermolecular C=O.H C close contacts are found. PMID- 11025319 TI - Benzyl 5-carboxy-4-ethyl-3-methylpyrrole-2-carboxylate. AB - In the title compound, C(16)H(17)NO(4), the benzyloxycarbonyl group is anti to the pyrrolic N atom. The molecules are joined into head-to-head dimers by hydrogen bonds involving the carboxylic acid groups. There is orientational disorder of these groups over two positions with approximately equal occupancy. A weaker hydrogen bond between the pyrrolic N atom and the carbonyl O atom of the benzyloxycarbonyl group joins the dimers into chains running parallel to the [110] direction. PMID- 11025321 TI - Bis(3-nitrophenyl) disulfide forms no C-H.O hydrogen bonds AB - In the title compound, C(12)H(8)N(2)O(4)S(2), the molecules lie across twofold rotation axes in the space group C2/c. There are no intermolecular C-H.O hydrogen bonds, but the molecules are linked into chains along [001] by aromatic pi.pi stacking interactions. PMID- 11025320 TI - Terbogrel, a dual-acting agent for thromboxane receptor antagonism and thromboxane synthase inhibition. AB - Terbogrel, (E)-6-[4-(3-tert-butyl-2-cyanoguanidino)phenyl]-6-(3-pyridyl)hex-5 enoic acid, C(23)H(27)N(5)O(2), a mixed thromboxane A(2) receptor antagonist and thromboxane A(2) synthase inhibitor, shows a hairpin-like conformation stabilized by an intramolecular hydrogen bond. A structural feature characteristic of the thromboxane A(2) synthase inhibitor mode is observed: a distance of 8.4257 (19) A between the pyridine N atom and the carboxyl group. PMID- 11025322 TI - (5S)-3-oxo-4-oxa-endo-tricyclo[5.2.1.0(2,6)]dec-8-en-5-yl acetate. AB - The molecular structure of C(11)H(12)O(4), based on a norbornene core, was established to confirm the configuration of an acetoxy side-chain group in addition to the formation of the endo product. The acetoxy side chain lies in an axial position relative to the five-membered fused ring. Bond distances and angles show no unusual features, with all geometric parameters lying within their expected ranges. The overall stereochemistry of the molecule was ascertained from the chiral furanone starting material. PMID- 11025323 TI - 2,4,6-Trichlorophenylisonitrile and 2,4,6-trichlorobenzonitrile AB - The molecular structures of the title compounds, 2,4, 6-trichlorophenylisonitrile (IUPAC name: 2,4,6-trichlorophenyl isocyanide), C(7)H(2)Cl(3)N, and 2,4,6 trichlorobenzonitrile, C(7)H(2)Cl(3)N, are normal. The two structures are not isomorphous, but do contain similar two-dimensional layers in which pairs of molecules are held together by pairs of Cl.CN [3.245 (3) A] or Cl.NC [3.153 (2) A] interactions. The two-dimensional isomorphism is lost through different layer stacking modes. PMID- 11025324 TI - DL-arginine monohydrate at 100 K. AB - In the title compound, C(6)H(14)N(4)O(2).H(2)O, the alpha-amino group is neutral. The molecular side chain including the guanidinium group is not fully extended, having a near gauche-gauche conformation [chi(3) = 59.0(1)degrees; chi(4) = 72.8(1)degrees]. The network of hydrogen bonds stabilizing the crystal lattice includes those formed between the deprotonated and negatively charged alpha carboxylate groups and the positively charged amino groups of the guanidinium group of neighbouring molecules. N-H...O=C and water-mediated N-H...O hydrogen bonds link individual molecules to produce pairs of spiral motifs laterally connected by N-H...O and C-H...O hydrogen bonds. PMID- 11025325 TI - A synchrotron diffraction study of two polymorphic forms of 2, 5-bis AB - Two polymorphs, alpha and gamma, of the title compound, C(22)H(20)N(2)O(2), have been characterized by means of single-crystal synchrotron X-ray diffraction. In the alpha form, the molecules pack in a herring-bone fashion, linked via weak C H.N intermolecular interactions (H.N 2.58 A). In the gamma form, the molecules are arranged in nearly planar sheets, which form a network held together by intermolecular hydrogen bonds of the type C-H.O (H. O 2.49 A) and C-H.N (H.N 2.50 A). The stacking distance between the sheets is 3.40 A. PMID- 11025326 TI - 2-Hydroxy-5- AB - The title compound, C(14)H(12)N(2)O(4), shows an E conformation about the diazenyl N atoms. The crystal structure features layers of molecules with the primary connection between the layers afforded by carboxylic acid dimer motifs; no evidence for extensive pi-pi stacking between the layers was found. PMID- 11025327 TI - An inhibitor of Janus kinase 3:4-(4-hydroxyphenylamino)-6, 7-dimethoxyquinazolin 1-ium chloride methanol solvate. AB - The crystal structure of the title compound, C(16)H(16)N(3)O(3)(+). Cl(-).CH(4)O (WHI-P131, an inhibitor of Janus kinase 3), contains four hydrogen bonds. There are two hydrogen bonds within the asymmetric unit, i.e. interactions between WHI P131 OH and Cl(-), and between methanol and Cl(-). There is a third interaction between WHI-P131 NH and Cl(-) (related by a 2(1) screw) and a fourth between WHI P131 NH and methanol (related by an n-glide). The hydrogen-bond pattern for these interactions can be described by the first-level hydrogen-bond graph-set notation D(1)(1)(2)D(1)(1)(2)D(1)(1)(2)D(1)(1)(2). The second-level graph-set notation (for combinations of two hydrogen bonds) was determined to be D(1)(2)(3)D(1)(2)(3)D(2)(2)(4)D(2)(2)(9)D(2)(2)(14)C(1)(2)(9). PMID- 11025328 TI - Oxythiamine hexafluorophosphate monohydrate, a thiamine antagonist with the same conformation as thiamine. AB - In the title compound, 3-[(3,4-dihydro-2-methyl-4-oxopyrimidin-5-yl) methyl]-5-(2 hydroxyethyl)-4-methylthiazolium hexafluorophosphate monohydrate,C(12)H(16)N(3)O(2)S(+).PF(6)(-).H(2)O, oxythiamine is a monovalent cation with a neutral oxopyrimidine ring. The molecule assumes the F conformation, which is a common form for thiamine but which is substantially different from the unusual V conformation found in the chloride and hydrochloride salts of oxythiamine. The anion-bridging interaction, C-H...anion...pyrimidine, is emphasized as being important for stabilization of the F conformation. PMID- 11025329 TI - Hydrogen bonding and C-H.O interactions in 4-phenanthrenemethanol at 150 K AB - The title compound, C(15)H(12)O, crystallizes in the centrosymmetric space group I4(1)/a with one molecule in the asymmetric unit. In the single hydrogen bond, the H atom is ordered, the O(D).O(A) distance is 2.788 (1) A and the O-H.O angle is 176 (1) degrees. Each hydroxyl group forms hydrogen bonds with two other hydroxyl groups and the resulting chains of interactions, in four non-linked subsets of molecules, propagate along [001]. The single leading intermolecular C H.O interaction has an H.O distance of 2.81 A and a C-H.O angle of 140 degrees; the single leading intramolecular C-H.O interaction has an H.O distance of 2.24 A and a C-H.O angle of 152 degrees. The phenanthrene core is less nearly planar in this structure than in the room temperature structure of phenanthrene-4 carboxylic acid. PMID- 11025330 TI - Acute and chronic effects of alcohol on preattentive auditory processing as reflected by mismatch negativity. AB - Chronic alcoholism, a major worldwide health problem, is associated with a variety of neurocognitive changes in the afflicted individuals. The precise neurophysiological basis of these changes is not yet understood. Mismatch negativity (MMN) is a preattentive event-related potential component indexing cortical auditory memory traces and automatic change detection in the brain that can be used to study the neural basis of cognitive impairments in various neurodegenerative diseases. MMN studies have revealed that even a low dose of acute alcohol significantly impairs automatic change detection and involuntary attention shifting. Recent MMN results on chronic alcoholism in turn suggest that auditory sensory traces decay slightly faster and are substantially more vulnerable to the distracting effect of backward masking in alcoholics than in healthy subjects. Furthermore, chronic alcohol abuse might accelerate the age related impairment of automatic change detection. There is also evidence that the MMN changes might predict impaired performance in behavioral memory and attention tasks in alcoholics. In MMN studies of detoxified alcoholics, however, many confounding factors have to be taken into account. For instance, postwithdrawal brain hyperexcitability might be associated with a slightly enhanced or accelerated MMN/MMNm (the magnetic equivalent of MMN). In sum, MMN and MMNm provide an objective noninvasive tool for exploring the neurophysiological functional deficits related to both acute alcohol intoxication and chronic alcoholism. PMID- 11025331 TI - Human frequency-following response: representation of tonal sweeps. AB - Auditory nerve single-unit population studies have demonstrated that phase locking plays a dominant role in the neural encoding of steady-state speech sounds. Recently, we have reported that the phase-locked activity underlying the human frequency-following response (FFR) could also encode the first two formants of several tonal approximations of steady-state vowels. Since auditory nerve single-unit population studies have also demonstrated that phase locking is used to represent time-varying speech-like sounds, it was reasoned that the phase locked neural activity underlying the human FFR, likewise, is dynamic enough to represent time-varying sounds. FFRs to a rising and a falling tone were obtained from 8 normal-hearing adults at 95, 85, 75 and 65 dB nHL. Results clearly demonstrated that the human FFR does indeed follow the trajectory of the rising and falling tones. Also, amplitude changes in the FFR supported the view that neural phase locking decreases with increasing frequency. Finally, the relatively smaller FFR amplitude for the falling tone compared to its rising counterpart lends further support to the notion that rising tones produce greater neural synchrony than falling tones. These results indicate that the human FFR may be used to evaluate encoding of time-varying speech sounds like diphthongs and certain consonant-vowel syllables. PMID- 11025332 TI - Auditory evoked response test strategies to reduce cost and increase efficiency: the postauricular muscle response revisited. AB - We describe a number of techniques in auditory evoked response (AER) testing for hearing loss which should decrease its cost and increase its efficiency, making its use in infant hearing screening more viable. We demonstrate the use of bit stream averaging of the electrical signals from the head as a cheap alternative to analogue averaging and show that the average waveforms obtained are similar with both techniques. We demonstrate how the postauricular muscle response (PAMR) can be potentiated by lateral rotation of the eyes and argue that uncontrolled eye movements in previous studies have led to an unfounded belief that the PAMR is not stable. When eye rotation is used to potentiate the PAMR, the response becomes very stable and so large in most subjects that it is clearly visible in the raw traces. We also demonstrate that when the PAMR is potentiated by eye rotation, stable PAMR waveforms can be reliably obtained with tone bursts with frequencies up to and above 8 kHz and with sound levels within 30 dB of the subjective detection threshold. As a result the PAMR can be used to rapidly determine an objective audiogram in most subjects within minutes. Finally, we demonstrate a correlation technique for detecting the PAMR without waveform averaging and the need for an expensive computer. We are sure that a combination of these techniques can be used to increase the efficiency of AER screening for infant deafness and lower its cost dramatically. PMID- 11025333 TI - A simple two-component model of the electrically evoked compound action potential in the human cochlea. AB - Neural response telemetry (NRT) permits in situ intracochlear recordings of the electrically evoked compound action potential from the auditory nerves using scala tympani electrodes. The recorded NRT waveforms can generally be categorized under either single positive peak or double positive peak waveforms. This is similar to the observations from Stypulkowski and van den Honert, who suggested that the double peak complex arises from two components that could be axonal and dendritic in origin, respectively. Using a simple mathematical model which linearly combines two separate waveforms similar in shape but differing in amplitude and latencies, it was possible to simulate the various NRT waveform categories. The simulation results support the view that the two waveform components originate from dendritic or axonal processes and implies that the shape of the response waveform may provide information about the degree of neural survival in the stimulated cochlea. This information could be useful for determining optimal speech coding parameters for cochlear implant users on an individual basis. PMID- 11025334 TI - Influence of the temperature signal on sleep in mammals. AB - The influence of the temperature signal on sleep may be considered physiologically specific if it entails thermoreceptor activation. Experimental evidence shows that sleep time peaks at neutral ambient temperature. PMID- 11025335 TI - The amygdala: a critical modulator of sensory influence on sleep. AB - The influence of external stimuli and the memories of both unpleasant and pleasant conditions clearly can have a considerable impact on the quality of sleep. The amygdala, a structure that plays an important role in coding the emotional significance of stimuli and is heavily interconnected with brainstem nuclei known to be involved in sleep control, has received little attention from sleep researchers. We report on a series of studies, focusing on its central nucleus (Ace). Presence of serotonin (5-HT) in Ace caused a rapid change of state when injected in rapid- eye-movement sleep (REM) compared with non-REM (NREM) injections. A 5-HT antagonist released ponto-geniculo-occipital waves (PGO) into NREM. Stimuli conditioned by pairing with aversive stimuli in a fear-conditioning paradigm significantly increased sound-elicited PGO and reduced REM. PMID- 11025336 TI - Reciprocal actions between sensory signals and sleep. AB - To the best of our knowledge, there is no simple way to induce neural networks to shift from waking mode into sleeping mode. Our best guess is that a whole group of neurons would be involved and that the process would develop in a period of time and a sequence which are mostly unknown. The quasi-total sensory deprivation elicits a new behavioral state called somnolence. Auditory stimulation as well as total auditory deprivation alter sleep architecture. Auditory units exhibiting firing shifts on passing to sleep (augmenting or diminishing) are postulated to be locked to sleep-related networks. Those ( approximately 50%) that did not change during sleep are postulated to continue informing the brain as in wakefulness. A rhythmic functional plasticity of involved networks is postulated. A number of auditory and visual cells have demonstrated a firing phase locking to the hippocampal theta rhythm. This phase locking occurs both during wakefulness and sleep phases. The theta rhythm may act as an organizer of sensory information in visual and auditory systems, in all behavioral states adding a temporal dimension to the sensory processing. Sensory information from the environment and body continuously modulates the central nervous system activity, over which sleep phenomenology must develop. It also produces a basal tonus during wakefulness and sleep, determining changes in the networks that contribute to sleep development and maintenance and, eventually, it also leads to sleep interruption. PMID- 11025337 TI - Bright light during nighttime: effects on the circadian regulation of alertness and performance. AB - The present studies evaluated to what extent duration (all-night or 4-hour exposures) and timing of nocturnal bright light (BL) (beginning or end of the night) modulate effects on vigilance. The results showed that all-night BL exposure is able to alleviate the nocturnal decrements in alertness and performance. However, under certain circumstances, this continuous BL exposure may induce adverse effects on mood and finally reveal to be counterproductive. Shorter BL exposure (4 h) during nighttime helps improve mood and performance, although the effects of short BL pulses were less efficacious than all-night BL exposure. The latter part of the night appears the best time for using the alerting effect of BL. The immediate alerting effect of BL seems to be mediated by a global activation of the central nervous system. PMID- 11025338 TI - Adenosine as a biological signal mediating sleepiness following prolonged wakefulness. AB - Recent reports from our laboratory have shown that extracellular adenosine levels selectively increase in basal forebrain during prolonged wakefulness in cats and rats. Furthermore, microdialysis perfusion of adenosine into the basal forebrain (BF) increased sleepiness and decreased wakefulness in both the species, whereas perfusion of the A(1)-receptor-selective antagonist, cyclopentyl-1, 3 dimethylxanthine resulted in increased wakefulness, an observation similar to that found with caffeine or theophylline administration. The selective participation of the A(1) subtype of the adenosine receptor in mediating the effects of adenosine in the BF was further examined by the technique of single unit recording performed in conjunction with microdialysis perfusion of selective agonists and antagonists. Perfusion of the A(1) agonist cyclohexyladenosine, inhibited the activity of wake-active neurons in the basal forebrain. The effect of prolonged wakefulness-induced increases in adenosine levels were further investigated by determining the changes in the BF in the levels of A(1) receptor binding and the levels of its mRNA. We observed that A(1) receptor mRNA levels increase after 6 h of sleep deprivation. One of the transcription factors that showed increased DNA-binding activity was nuclear factor kappaB (NF-kappaB) and may regulate the expression of A(1) mRNA. We observed, using a gel shift assay, that the DNA-binding activity of NF-kappaB increased following 3 h of sleep deprivation. This was further supported by the increased appearance of NF-kappaB protein in the nuclear extracts and the consequent disappearance of cytoplasmic protein inhibitor kappaB (I-kappaB). Together our results reviewed in this report suggest that the somnogenic effects of adenosine in the BF area may be mediated by the A(1) subtype of adenosine receptor, and its expression might be regulated by induction in the NF-kappaB protein as its transcription factor. This positive feedback might mediate some of long-duration effects of sleep deprivation, including 'sleep debt'. PMID- 11025339 TI - A critical assessment of the melatonin effect on sleep in humans. AB - Melatonin is synthesized and secreted during the dark period of the light-dark cycle. The rhythmic nocturnal melatonin secretion is directly generated by the circadian clock, located in mammals within the suprachiasmatic nucleus (SCN), and is entrained to a 24-hour period by the light-dark cycle. The periodic secretion of melatonin may be used as a circadian mediator to any system that can 'read' the message. In addition, direct effects of the hormone on the SCN could explain some of the melatonin effects on the circadian system. Duration of the melatonin nocturnal secretion is directly proportional to the length of the night and it has experimentally been demonstrated to be the critical parameter for photoperiod integration. The sites and mechanisms of action of melatonin for circadian and photoperiodic responses are far from being elucidated, but action through specific membrane receptor sites starts to emerge. A possible bicompartmental model of distribution for melatonin, the first compartment in plasma acting on peripheral organs and the second in the cerebrospinal fluid affecting neurally mediated functions at a much higher concentration, has recently been proposed. From earlier studies it was concluded that melatonin administration to humans reduces sleep latency and induces sleepiness and fatigue. More recently, the effect of lower pharmacologic or physiologic doses of melatonin was examined in different laboratories. These studies included young normal volunteers and patients with chronic insomnia, as well as dementia patients exhibiting sundowning syndrome. Irrespective of the method of assessment, melatonin showed effects in insomniac patients in most studies. With some exceptions, melatonin administration reduced sleep latency and/or increased total sleep time and sleep efficiency. Furthermore, melatonin was more effective when given to elderly insomniacs, or Alzheimer disease patients, although sleep improvement was not strictly correlated with prior levels of the hormone. PMID- 11025340 TI - Seasonal cell proliferation in the chemosensory epithelium and brain of red backed salamanders, Plethodon cinereus. AB - The chemosensory epithelium of vertebrates retains the ability to produce new receptor neurons throughout life, presumably as a mechanism to replace aging or damaged receptors. We examined cell division in the main olfactory and vomeronasal epithelia of red-backed salamanders (Plethodon cinereus) because previous studies had shown that the volume of sensory epithelia changes seasonally. Cell division was compared throughout the year by injecting salamanders once with 5-bromo-2'-deoxyuridine (BrdU), which is incorporated into the DNA of cells during DNA synthesis, and sacrificing them one hour after injection. We used immunocytochemistry to locate cells that had arisen from cell division since BrdU injection and compared the number of labeled cells per area among animals. Animals collected in May had significantly more labeled nuclei than animals collected in any other month. However, proliferation rates among the other months were not significantly different and were quite low. Labeled nuclei also were found around the cerebral ventricles of salamanders collected in May, but rarely in any other month, although other tissues in the head often were heavily labeled. Cell proliferation appears to be up-regulated in the chemosensory epithelia and in the telencephalon during May, and we hypothesize that new receptors, and perhaps their interneurons in the telencephalon, are being generated in anticipation of seasonal events that are mediated by chemoreception. PMID- 11025341 TI - Sonic/vocal motor pathways in squirrelfish (Teleostei, Holocentridae). AB - Similar to many teleost fish, squirrelfish (family Holocentridae) produce vocalizations by the contraction of muscles that lead to vibration of the swimbladder. We used biotinylated compounds to identify the position and extent of vocal motor neurons in comparison to additional motor neuron groups, namely those of red and white dorsal epaxial muscle and opercular muscle that are located adjacent to or near the sonic muscle. The sonic motor nucleus (SMN) was located in the caudal medulla and rostral spinal cord in a ventrolateral position with dendrites extending dorsally in a dense bundle along the lateral edge of the medulla and axons exiting via ventral occipital nerve roots. Transneuronal transport of biocytin identified premotor neurons within the SMN and in the medially adjacent reticular formation that projected to the contralateral SMN and more rostrally to the octavolateralis efferent nucleus and nucleus praeeminentialis, suggesting interactions between vocal and octavolateralis systems as seen in other teleosts. Motor neurons innervating the red and white dorsal muscle formed a loose aggregate in the dorsal motor column, adjacent to the medial longitudinal fasciculus, sending fibers bilaterally throughout the spinal cord with axons exiting via ventral spinal nerve roots. Opercular motor neurons were located within the facial motor nucleus. The anatomical characteristics of the SMN of squirrelfish, a representative member of the order Beryciformes, are similar to those of representative members of the closely related order Scorpaeniformes, but diverge from the SMN of more distantly related orders of paracanthopterygiian and ostariophysan teleosts. These results therefore suggest a possible homology among the SMNs of acanthopterygiian fishes. PMID- 11025342 TI - Telencephalic aromatase but not a song circuit in a sub-oscine passerine, the golden collared manakin (Manacus vitellinus). AB - In oscine passerines, the telencephalon expresses high levels of the estrogen synthetic enzyme aromatase. In contrast, forebrain aromatase is limited to low levels at discrete limbic loci in non-passerines. The function of forebrain aromatase in oscines is unknown, however, estrogen-sensitive elements of the telencephalic song circuit (an oscine characteristic) may be influenced by local aromatization. Very few studies have investigated the neuroendocrine characteristics of sub-oscine passerines. Species of this passerine sub-order are taxonomically similar to oscines, but do not appear to learn how to sing as oscines, and show no evidence of a song circuit. The neural expression of aromatase in these birds is unknown. We asked whether the golden-collared manakin, a sub-oscine, (a) showed evidence of a song circuit, and (b) expressed aromatase in the telencephalon at high levels like the zebra finch (oscine passerine) or at low levels like the quail (non-passerine). Nissl stains and immunocytochemistry for microtubule associated proteins showed no evidence of a song circuit in manakins of either sex, whereas both techniques delineate all song nuclei in the zebra finch. However, biochemical and immunocytochemical measures reveal that in the manakin, several telencephalic loci, including the hippocampus, caudomedial neostriatum, nucleus taeniae, and the lateral neostriatum express aromatase. Assays run in parallel show low to undetectable levels of aromatase in the telencephalon of the quail (nonpasserine) and abundant levels in the zebra finch (oscine passerine), suggesting a dissociation between the presence of a song circuit and forebrain aromatase expression in this sub oscine. These data suggest that forebrain aromatase may have evolved in sub oscine songbirds before the evolution of a song circuit and singing behavior in oscines. Alternatively, forebrain aromatase may serve functions distinct from singing behavior. PMID- 11025343 TI - Early histological maturation in the hippocampus of the guinea pig. AB - The vesicular zinc-rich synaptic systems of the principal neurons of the hippocampus are well developed in newborn guinea pigs, a precocial species. In addition, alvear and fimbrial myelinated fibers as well as significant inhibitory interneurons (i.e. somatostatin, parvalbumin and opioid immunoreactive hippocampal interneurons) are also well developed. On the contrary, neither vesicular zinc synapses nor myelinated fibers nor the above mentioned immunoreactive interneurons are detectable in newborn specimens of other related altricial species such as rats or rabbits. These data suggest that early maturation of a highly integrative center related to cognitive map building such as the hippocampus is characteristic of precocial species. PMID- 11025344 TI - Signal decoding and receiver evolution. An analysis using an artificial neural network. AB - We use a connectionist model, a recurrent artificial neural network, to investigate the evolution of species recognition in sympatric taxa. We addressed three questions: (1) Does the accuracy of artificial neural networks in discriminating between conspecifics and other sympatric heterospecifics depend on whether the networks were trained only to recognize conspecifics, as opposed to being trained to discriminate between conspecifics and sympatric heterospecifics? (2) Do artificial neural networks weight most heavily those signal features that differ most between conspecifics and sympatric heterospecifics, or those features that vary less within conspecifics? (3) Does selection for species recognition generate sexual selection? We find that: (1) Neural networks trained only on self recognition do not classify species as accurately as networks trained to discriminate between conspecifics and heterospecifics. (2) Neural networks weight signal features in a manner suggesting that the total sound environment as opposed to the relative variation of signals within the species is more important in the evolution of recognition mechanisms. (3) Selection for species recognition generates substantial variation in the relative attractiveness of signals within the species and thus can result in sexual selection. PMID- 11025345 TI - Exercise-induced ST segment elevation in Q wave leads in postinfarction patients: defining its meaning and utility in today's practice. AB - Most attempts to identify qualitative and quantitative techniques for assessing myocardial viability and the likelihood of improved function after revascularization in patients with healed myocardial infarcts have focused on treatment strategies and prognosis. This review examines the true value of the electrocardiographic phenomenon of exercise-induced ST segment elevation (EISTE) in Q wave leads as a diagnostic tool for the assessment of myocardial viability. The prognostic potential and clinical utility of the EISTE phenomenon are inhibited both by the heart's electrophysiologic response to exercise-induced metabolic and hemodynamic changes, and by the ECG's limited facility in assessing myocardial preservation. The use of EISTE as an independent indicator for surgical intervention is proscribed by these limitations. The EISTE phenomenon could serve as a useful tool in the first line of discrimination in patients with healed Q wave myocardial infarction, and may justify further diagnostic work-up in patients under consideration for a revascularization procedure. In the era of sophisticated nuclear and echo techniques, accurate imaging studies should not be replaced by ECG analysis alone in the search for viable tissue, except when financial costs are of major importance. PMID- 11025346 TI - Update on the management of acute coronary syndromes. AB - At present there is debate as to whether an invasive or a conservative strategy should form the basis of an optimal management strategy for unstable angina/non-Q wave myocardial infarction (UA/NQMI). However, these approaches are complementary, not necessarily mutually exclusive. On the basis of current evidence, all patients should receive optimized medical therapy, with surgical interventions targeted at high-risk patients, to improve both clinical outcomes and cost effectiveness. While standard antithrombotic combinations have improved short-term outcomes, they do not fully eliminate the risk of recurrent ischemic episodes. The recent introduction of direct thrombin inhibitors, platelet fibrinogen receptor antagonists and low-molecular-weight heparins (LMWHs) has offered an opportunity to develop more aggressive antithrombotic regimens. Enoxaparin, an LMWH, has demonstrated improved efficacy compared with standard heparin in both the acute and chronic phases of UA/NQMI, without an increase in major complications caused by bleeding. Further studies are justified to investigate the potential of combined antithrombotic regimens containing enoxaparin as an alternative to heparin in conservative strategies and as adjuncts to interventional procedures. Recommendations for the management of UA/NQMI should be continually reviewed in response to the impact of novel treatment modalities. PMID- 11025347 TI - Infiltrative nonamyloidotic monoclonal immunoglobulin light chain cardiomyopathy: an underappreciated manifestation of plasma cell dyscrasias. AB - BACKGROUND: Infiltrative cardiomyopathies are characterized by diastolic dysfunction. In monoclonal plasma cell dyscrasias, organ compromise may be produced by tissue deposition of monoclonal immunoglobulins or their constituent peptides independently of the effects of unbridled plasma cell proliferation. The deposits may be fibrillar, as in light chain amyloid (AL) or nonfibrillar, as in light chain deposition disease (LCDD). AL disease of the heart is a restrictive cardiomyopathy. We hypothesized that, despite differences in physical properties, nonamyloidotic light chain deposition in the myocardium could produce similar clinical and physiological abnormalities. METHODS: Cardiac tissue from five patients with LCDD and cardiac dysfunction was examined by immunohistochemical and electron microscopic techniques. Hospital charts, electrocardiograms, echocardiograms and cardiac catheterization results were reviewed. In two cases, the original echocardiograms were reanalyzed. RESULTS: The five patients with nonamyloidotic light chain deposits in the myocardium had either mechanical or electrocardiographic abnormalities. In four with adequate clinical documentation, the diastolic dysfunction and conduction abnormalities were similar or identical to that described in cardiac AL disease. CONCLUSIONS: Although nonamyloidotic immunoglobulin light chain deposits in the myocardium differ in distribution and ultrastructural organization from the fibrillar deposits of AL disease, an analogous pattern of diastolic dysfunction and conduction disturbances results. The diagnosis should be considered in patients with a plasmacytic dyscrasia and restrictive cardiomyopathy in whom Congo red staining of endomyocardial biopsy tissue is negative. The diagnosis can be established by using the appropriate immunohistochemical and ultrastructural tissue examinations. PMID- 11025348 TI - Left ventricular structure and function in primary hyperparathyroidism before and after parathyroidectomy. AB - OBJECTIVE: Our aim was to study the effect of primary hyperparathyroidism (PHPT) and parathyroidectomy (PTX) on left ventricular (LV) wall thicknesses and systolic and diastolic function. METHODS: Fifteen patients with untreated PHPT were evaluated by applying Doppler and digitized M-mode echocardiography before and 2-3 months after PTX. Fifteen age- and sex-matched healthy controls were also examined echocardiographically. RESULTS: Prior to PTX, interventricular septal thickness (IVST), LV mass (LVM), aortic root dimension and left atrium dimension were greater and LV fractional shortening was slightly decreased in patients as compared to controls. Significantly increased LV peak late diastolic velocity (A(max)) and isovolumic relaxation time, and a slightly decreased ratio of peak early to peak late diastolic velocities (E/A(max)) in the patients indicated impairment of LV diastolic function in hyperparathyroidism. PTX reduced serum total Ca from 2. 79 +/- 0.13 to 2.39 +/- 0.09 mmol/l (p < 0.001) and tended to reduce IVST [10.6 +/- 2.1 vs. 10.4 +/- 2.0 mm; not significant (n.s.)], LV posterior wall thickness (9.6 +/- 2.0 vs. 9.2 +/- 1.0 mm, n.s.) and LVM (250 +/- 102 vs. 213 +/- 42 g; n.s.). Before PTX, there was a significant correlation between serum total Ca and LVM (r = 0.63, p < 0.05), and the PTX-induced change in serum total calcium correlated with the change in LVM (r = 0.59, p < 0.05). PTX induced no significant changes in LV systolic or diastolic function during the follow-up of 2-3 months. CONCLUSIONS: The present findings indicate that PHPT induces LV hypertrophy, slight impairment of LV systolic function and significant impairment of LV diastolic function, which are not substantially improved after TX and 2-3 months of normocalcemia. PMID- 11025349 TI - Temperature-controlled high frequency ablation for creation of transmyocardial channels: in vivo validation of a novel method. AB - OBJECTIVE: We investigated the feasibility and short-term effects of a novel procedure to create intramyocardial channels by means of high frequency (HF) ablation in a rabbit in vivo model. METHODS: A flexible catheter ending in a cylindrical electrode (diameter 0.7 mm) with a sharpened tip was used for HF energy application following transmyocardial insertion. Power-controlled or energy-controlled energy applications were performed in 16 anesthetized rabbits after thoracotomy with a follow-up for 3 h. Assessment of myocardial channels and the necrotic zone was performed by morphometric quantification in serial sections. The ferret diameter was used to compare channel dimensions and the extent of necrosis. RESULTS: Thirty-nine power-controlled and 54 temperature controlled HF applications were performed. The shape of identified channels was round in 71% and 69% had a lumen patency of > or =2/3 of the channel. Ferret diameter of the channels was 414 +/- 180 microm and of the necrotic zone 3,558 +/ 1,200 microm. In temperature-controlled applications, channel dimensions were strongly influenced by the maximum tissue temperature and the duration of energy delivery (T(max): p = 0.0006; duration: p = 0. 003). Channel and necrosis dimensions correlated better with biometric parameters in temperature-controlled compared with power-controlled applications. CONCLUSION: Mechanically created transmyocardial channels can be stabilized by HF heating of the surrounding tissue. A high percentage of these channels remain patent. The channel dimensions are closely correlated with maximum temperature and duration of energy delivery in a temperature-controlled application mode. PMID- 11025350 TI - The utility of four biochemical markers in the triage of chest pain patients. AB - Four biochemical markers, creatine kinase (CK)-MB isoenzyme, myoglobin, myosin light chains and troponin I, were studied in 1,338 patients presenting to the emergency department with chest pain suggestive of coronary artery disease (CAD). One hundred and eighty-seven patients had an acute myocardial infarction (MI). At least one of the four markers was over the threshold on the first sample in 78% of MI patients, as compared to only 40% with an elevated CK-MB. After 4 h, 88% had at least one marker elevated. None of the 69 patients with atypical chest pain, no history of CAD, no markers over threshold on the first sample and a normal electrocardiogram had an acute MI or unstable angina. If we had discharged this group, we would have saved USD 264,000, estimating a cost of USD 2,000 per day. Using four biochemical markers improved the early diagnosis of CAD and may help identify groups suitable for early discharge. PMID- 11025351 TI - Cardiac troponin T levels at 96 hours reflect myocardial infarct size: a pathoanatomical study. AB - We determined the utility of single-point measurements of circulating cardiac troponin T (cTnT) for the noninvasive estimation of infarct size in 16 beagle dogs after left anterior descending artery (LAD) ligation. Pathoanatomical infarct sizes were determined by the triphenyltetrazolium chloride method and correlated with serum concentration changes of cTnT. Peak cTnT levels (14.10 +/- 4.71 microg/l) were reached after 110 +/- 21 h. A significant correlation was found between peak cTnT levels (p = 0.0001, r = 0. 83) or cumulative cTnT levels and relative infarct size (p = 0.0010, r = 0.72). A single cTnT measurement 96 h after LAD ligation was equally predictive of infarct size (p = 0.0010, r = 0.74) as peak or cumulative cTnT levels derived from serial sampling. cTnT levels at 96 h may thus be useful for practical and cost-effective estimation of infarct size. PMID- 11025352 TI - Identification of markers of different severity of lesions in the proximal left main coronary artery. PMID- 11025353 TI - Potential uses of intravenous proton pump inhibitors to control gastric acid secretion. AB - Proton pump inhibitors are the most effective agents for suppressing gastric acidity and are the preferred therapy for many acid-related conditions. While proton pump inhibitors have been accessible in intravenous formulations in several European countries, they have been available only as oral drugs in the United States. In the near future, the proton pump inhibitor pantoprazole is likely to become available in an intravenous formulation for American patients. Potential uses for intravenous proton pump inhibitors include treatment of Zollinger-Ellison syndrome and peptic ulcers complicated by bleeding or gastric outlet obstruction, as well as prevention of stress ulcers and acid-induced lung injury. These intravenous proton pump inhibitors are also likely to be beneficial to patients undergoing long-term maintenance with oral proton pump inhibitors who cannot take oral therapy for a period of time. Intravenous pantoprazole is especially distinguished in its lack of clinically relevant drug interactions, and it requires no dosage adjustment for patients with renal insufficiency or with mild to moderate hepatic dysfunction. Both omeprazole and pantoprazole are well tolerated in both oral and intravenous forms. Although further studies are needed to define their roles clearly, the availability of intravenous formulations of proton pump inhibitors will certainly assist with the treatment of gastric acid-related disorders. PMID- 11025354 TI - Helicobacter pylori and gastric erosions. Results of a prevalence study in asymptomatic volunteers. AB - BACKGROUND/AIMS: Helicobacter pylori is considered to be the primary cause of most forms of gastritis, but its role as a causative agent in gastric erosions is unclear. The aim of this study was to estimate the prevalence of gastric erosions and H. pylori infection in asymptomatic volunteers. METHODS: 175 asymptomatic subjects underwent upper gastrointestinal endoscopy. Antral biopsies were taken for bacterial cultures, histology and quick urease (CLO) test. A (13)C-urea breath test was performed after endoscopy. NSAID intake, alcohol consumption and smoking habits were also recorded in each subject. RESULTS: 33 (19%) of 175 asymptomatic volunteers had macroscopic lesions on upper gastrointestinal endoscopy, 7 were H. pylori positive, 26 were H. pylori negative. Gastric erosions occurred in 8% (14 subjects) of all volunteers. 10 subjects were H. pylori negative and 4 H. pylori positive. In 11 volunteers, gastric erosions were restricted to the prepyloric antrum. Only 1 of 14 subjects had a history of NSAID intake and 6 subjects were alcohol abstainers. CONCLUSION: We conclude that gastric erosions occur in a considerable amount of asymptomatic volunteers. They are predominantly localized in the prepyloric antrum and are most likely not associated with H. pylori infection, NSAID intake, smoking or alcohol consumption. PMID- 11025355 TI - Helicobacter pylori-associated gastric ulcer exhibits enhanced mucosal chemokine activity at the ulcer site. AB - BACKGROUND AND AIM: Although mucosal alpha- and beta-chemokines are considered to be involved in the pathogenesis of Helicobacter pylori-associated gastritis, little is known how these chemokines are related to the ulcerogenesis in peptic ulcer patients. We examined the levels of interleukin (IL)-8 and macrophage inflammatory protein-1alpha (MIP-1alpha) in organ cultures and the numbers of inflammatory cells infiltrating the lamina propria by using the mucosal tissues obtained from gastric ulcer (GU) patients with and without H. pylori infection. METHODS: Levels of IL-8 and MIP-1alpha secreted in organ cultures were measured by an enzyme-linked immunosorbent assay. Numbers of myeloperoxidase-positive neutrophils, CD68-positive macrophages, and mononuclear cells were determined in tissue sections. RESULTS: The mucosal tissues of both the gastric antrum and the ulcer site obtained from patients with H. pylori-positive GU showed significantly higher levels of IL-8 and MIP-1alpha and increased numbers of inflammatory cells compared with the corresponding mucosal tissues from those with H. pylori negative GU or the antral mucosal tissues from H. pylori-negative controls. When the values were compared between the mucosal tissues from the gastric antrum and those from the ulcer site, the latter group of tissues showed significantly higher levels of IL-8 and MIP-1alpha and increased numbers of neutrophils and macrophages than the former group regardless of its healing process in patients with H. pylori-positive GU. CONCLUSION: Mucosal alpha- and beta-chemokines may be important to the ulcerogenesis in H. pylori-associated GU disease. PMID- 11025356 TI - Colonization with cagA-positive Helicobacter pylori strains inversely associated with reflux esophagitis and Barrett's esophagus. AB - AIM: The hypothesis that colonization with cagA(+) Helicobacter pylori strains protects against the development of gastroesophageal reflux disease (GERD) and its complications is tested. METHODS: Patients with reflux esophagitis and Barrett's esophagus were studied. Antral biopsy specimens were obtained for detection of H. pylori. A serum sample was obtained for determination of IgG antibodies to H. pylori and to the CagA protein. RESULTS: 736 patients were studied. 118 patients had reflux esophagitis, 36 had Barrett's esophagus, 108 had hiatal hernia without signs of inflammation (the reflux group), and 20 patients had esophageal or stomach cancer. The remaining 454 patients had no signs of GERD. The 262 patients with reflux disease had a significantly lower prevalence of H. pylori (34.9%) than the 454 controls (54.6%; p<0. 001). Among 310 H. pylori positive patients from whom serum was available, colonization with cagA(+) strains was detected in 59% in the control group versus 35% in the reflux group (p<0.001). CONCLUSION: Patients with reflux esophagitis and Barrett's esophagus have a significantly lower prevalence of H. pylori colonization than controls, in particular of the cagA(+) type. These data suggest that colonization with cagA(+) H. pylori strains may be protective against the development of GERD PMID- 11025357 TI - Water extracts of Helicobacter pylori suppress the expression of histidine decarboxylase and reduce histamine content in the rat gastric mucosa. AB - BACKGROUND: Acute Helicobacter pylori (Hp) infection in humans may be associated with markedly reduced gastric acid secretion, but the mechanism of this hypochlorhydria has not been fully explained. AIMS: This study was designed to investigate how water extracts (WE) of Hp applied on rat gastric mucosa affect gastric secretion and mucosal histamine concentration as well as the gene expression for histamine decarboxylase (HDC), the key enzyme converting histidine to histamine and for interleukin-1beta (IL-1beta), the important proinflammatory cytokine. MATERIALS AND METHODS: Wistar rats were surgically equipped with small cannulas to form gastric fistulas (GF). Four weeks after formation of GF, rats received either saline (control group) or WE obtained from type I Hp strain expressing CagA/VacA proteins and from type II Hp strain negative for CagA/VacA. Hp-WE was applied intragastrically (i.g.) in a volume of 1 ml at days 0, 2, 4 and 6 (total 4 times). At days 7 and 14, the secretory tests were performed during which basal gastric acid and pepsin secretion was examined and acid and pepsin outputs were measured. After secretory tests, the rats were sacrificed, the stomachs removed and the damage to the gastric mucosa was assessed by measuring the lesion area planimetrically and by histology, the gene expression in gastric mucosa for HDC and interleukin-1beta (IL-1beta) was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot. Additionally, somatostatin concentration in gastric juice, gastric mucosal histamine content and plasma gastrin and IL-1beta levels were determined using radioimmunoassay (RIA). RESULTS: Administration of Hp-WE failed to induce gross mucosal damage but microscopic examination revealed partial denudation of gastric surface epithelium without causing deep necrosis. In secretory tests, Hp-WE produced marked hypochlorhydria but type I Hp-WE induced significantly stronger inhibition of acid and pepsin secretion than type II Hp-WE, both at days 7 and 14. Both, type I and type II Hp-WE suppressed significantly the gene expression for HDC mRNA and lowered significantly gastric mucosal histamine content as compared to respective values in vehicle-treated control gastric mucosa. Furthermore, Hp-WE, resulted in a significant increase in expression of IL-1beta mRNA and a significant fall in luminal somatostatin concentration as well as a insignificant elevation of plasma gastrin level, the type I Hp-WE being more effective in these alterations than type II Hp-WE. CONCLUSIONS: (1) Ability of Hp-WE to induce superficial damage, the reduction in HDC mRNA and accompanying fall in gastric histamine release, contribute, at least in part, to marked hypochlorhydria observed in the stomach exposed to repeated Hp-WE treatments, and (2) the deleterious effect of Hp-WE on the gastric mucosa involves an impairment of gastrin-somatostatin link possibly resulting from the action of Hp-derived toxins and the induction in mucosal cells of proinflammatory cytokine such as IL-1beta. PMID- 11025359 TI - Effect of ecabet disodium, a novel locally-acting antiulcer drug, on epithelial restitution following injury by hypertonic NaCl in bullfrog stomach in vitro. AB - BACKGROUND: The antiulcer drug ecabet 2Na (12-sulfodehydroabietic acid disodium salt) exhibits a gastroprotective activity, mainly through a local action, involving endogenous prostaglandins (PGs) and nitric oxide (NO). In the present study, we examined the effect of ecabet on the epithelial restitution of the bullfrog gastric mucosa in vitro following injury by hypertonic NaCl. METHODS: Bullfrog fundic mucosa was mounted in an Ussing chamber. The tissue injury was induced by exposure of the mucosa to 1.25 M NaCl for 5 min, and transmucosal potential difference (PD) and electrical resistance (R) were measured during a 4 hour test period. Ecabet (3-30 mg/ml) was added to the luminal solution for 10 min before or after NaCl, while 16,16-dimethyl PGE(2) (dmPGE(2): 1x 10(-6) M) or NOR-3 (a NO donor: 1 x 10(-4) M) was added to the nutrient solution 10 min before NaCl. RESULTS: Mucosal application of 1.25 M NaCl caused an immediate reduction of PD and R, followed by a gradual normalization, reaching about 70% of the pre exposure levels within 4 h. Ecabet, added before NaCl, significantly expedited the recovery of PD and R in a concentration-dependent manner; this effect was mimicked by posttreatment with ecabet and significantly mitigated by prior addition of indomethacin (1 x 10(-5) M) or N(G)-nitro-L-arginine methyl ester (L NAME: 1 x 10(-3) M). The epithelial restitution was also significantly promoted by serosal application of either dmPGE(2) or NOR-3. The mucosal exposure to ecabet significantly increased the luminal release of PGE(2) and NO metabolites, the effects being attenuated by indomethacin and L-NAME, respectively. The mucous secretion was increased by ecabet as well as dmPGE(2) and NOR-3, and the effect of ecabet was significantly suppressed by both indomethacin and L-NAME. The inhibitory effects of L-NAME on the ecabet action were all significantly antagonized by concurrent addition of L-arginine. CONCLUSION: These results suggest that ecabet significantly expedited the restitution following gastric surface cell injury, and this action is mediated by endogenous NO as well as PGs and may be functionally associated with an increase of mucous secretion. PMID- 11025358 TI - Effect of modified sham feeding and insulin-induced hypoglycemia on function of the proximal stomach. AB - BACKGROUND/AIM: Animal experiments have shown that vagal cholinergic stimulation causes an increase in proximal gastric tone, but little is known about the effect of vagal stimulation on proximal gastric motor function in humans. Vagal cholinergic stimulation can be elicited by modified sham feeding (MSF) or by insulin-induced hypoglycemia. The aim of our study was to investigate the effect of MSF and insulin-induced hypoglycemia on the motor and sensory function of the proximal stomach in humans. METHODS: Eight healthy volunteers participated in random order in three experiments: (A) control experiment, (B) MSF and (C) intravenous insulin injection. Intragastric volume was recorded with a barostat set at a constant preselected pressure level (MDP + 2 mm Hg). Pancreatic polypeptide (PP) secretion was measured as an indicator of cholinergic tone. RESULTS: PP secretion increased significantly after both MSF (p<0.05) and insulin administration (p<0.01). No changes in intragastric volume were seen after MSF, while intragastric volume increased significantly in response to insulin-induced hypoglycemia when compared to control (290+/-43 vs. 148+/-24 ml; p<0.01). No differences in perception scores were seen between the three experiments. CONCLUSIONS: Vagal cholinergic stimulation by MSF has no effect on the motor function of the proximal stomach, while insulin-induced hypoglycemia causes a relaxation of the proximal stomach. PMID- 11025360 TI - Central leptin and cholecystokinin in gastroprotection against ethanol-induced damage. AB - BACKGROUND: Leptin, a product of the ob gene controlling food intake, has recently been detected in the stomach and shown to be released by cholecystokinin (CCK) and to induce gastroprotection against various noxious agents, but it is not known whether centrally applied leptin influences gastric secretion and mucosal integrity. AIMS: In this study we compared the effects of leptin and CCK 8 applied intracerebroventricularly (i.c.v.) on gastric secretion and gastric mucosal lesions induced by topical application of 75% ethanol. METHODS: Several major series of Wistar rats were used in this study. The effects of leptin or CCK applied i.c.v. on gastric secretion were examined using conscious rats with gastric fistulas. For the studies on gastroprotection the following series of rats were used to determine the effects of: (A) leptin and CCK applied centrally on this protection and the blockade of CCK(A) with loxiglumide (30 mg/kg i.p.) and CCK(B) receptors with RPR 102681 (30 mg/kg i.p.); (B) cutting of vagal nerves; (C) inactivation of sensory nerves by capsaicin (125 mg/kg s.c.); (D) inhibition of calcitonin gene-related peptide (CGRP) receptors with CGRP(8-37) (100 microg/kg i.p.), and (E) suppression of nitric oxide synthase (NOS) with N(G)-nitro-L-arginine methyl ester (L-NAME) (5 mg/kg i.v. ) on ethanol-induced gastric lesions in rats with or without the i.c.v. pretreatment with leptin or CCK-8. Rats were anesthetized 1 h after ethanol administration to measure the gastric blood flow (GBF) and then to determine the area of gastric lesions by planimetry. Blood was withdrawn for the measurement of plasma leptin and gastrin levels by radioimmunoassay and gastric biopsy samples were collected for the determination of cNOS and iNOS mRNA by RT-PCR. RESULTS: Leptin and CCK-8 (0.01-5 microg/kg i.c.v.) dose dependently attenuated gastric lesions induced by 75% ethanol; the doses reducing these lesions by 50% (ED(50)) were 0.8 and 1.2 microg/kg, respectively. The protective effects of leptin and CCK-8 applied i.c. v. were accompanied by a significant rise in plasma leptin level and an increase in GBF. Blockade of CCK(A) receptors with loxiglumide abolished the protective and hyperemic effects of CCK but not those of leptin, while RPR 10268, a specific antagonist of CCK(B) receptors, counteracted leptin-induced protection and the rise in the GBF but failed to influence those afforded by CCK-8. For comparison, pretreatment with peripheral CCK-8 or leptin (10 microg/kg i.p.) causing a similar rise in the plasma leptin level also significantly reduced gastric lesions induced by 75% ethanol. The protective and hyperemic effects of centrally administered leptin were abolished by vagotomy, producing a fall in plasma leptin levels, and significantly attenuated by sensory denervation with capsaicin, by pretreatment with the CGRP antagonist, CGRP(8-37), or with L-NAME. A strong signal for iNOS mRNA was recorded in the gastric mucosa of leptin- and CCK-8 treated animals, whereas cNOS mRNA was unaffected. CONCLUSIONS: (1) Central leptin exerts a potent gastroprotective action at a dose that has no influence on gastric secretion; (2) this protection depends upon CCK(B) receptors, vagal activity and sensory nerves, and involves hyperemia probably mediated by NO, and (3) leptin mimics the gastroprotective effect of CCK and may be implicated in the protective and hyperemic actions of this peptide on the rat stomach. PMID- 11025361 TI - Gastrin-mediated alterations in gastric epithelial apoptosis and proliferation in a mastomys rodent model of gastric neoplasia. AB - BACKGROUND/AIMS: Hypergastrinemia secondary to low acid secretion is associated with gastric carcinoid formation in Mastomys. We investigated the effect of gastrin on oxyntic epithelial apoptosis and proliferation in this model. METHODS: Hypergastrinemia and mucosal hyperplasia were induced by irreversible H(2) receptor blockade with loxtidine. Gastrin levels were normalised in some animals by 10 days' loxtidine withdrawal. Serum gastrin was determined by radioimmunoassay, proliferative, enterochromaffin-like cells and Bcl-2 protein family expression by immunohistochemistry, and apoptotic cells by terminal deoxyuridine nucleotide nick end labeling (TUNEL). RESULTS: Proliferating cells were increased 4-fold in loxtidine-treated animals, and returned to normal upon loxtidine withdrawal. Enterochromaffin-like cell number increased 2-fold with loxtidine, and did not decrease after withdrawal. Apoptotic epithelial cells were located at the luminal surface and increased 1.8-fold with loxtidine, returning to control levels upon withdrawal. The ratio of proliferative to apoptotic cells was lower in the control and withdrawn groups than in the loxtidine group (0.26+/ 0.05 and 0.26+/-0.08 vs. 0.77+/-0.12). With hypergastrinemia, the expression of Bcl-2 and Bak was increased and Bax decreased in the middle of the gland. CONCLUSION: Hypergastrinemia is associated with alterations in both proliferation and apoptosis in Mastomys gastric mucosa. This may contribute to the pathogenesis of mucosal hyperplasia in this model. PMID- 11025362 TI - Substrate and inhibitor specificity of butyrate uptake in apical membrane vesicles of the rat distal colon. AB - BACKGROUND/AIMS: There is substantial evidence that transcellular flux of short chain fatty acid (SCFA) absorption, at least in part, is mediated by an anion exchange process with bicarbonate. METHODS: This anion exchange system was further characterized in apical membrane vesicles of the rat distal colon by studying substrate and inhibitor specificities of a variety of substituted monocarboxylic acids as well as of known inhibitors of the recently described monocarboxylate transporter MCT1 and MCT2. RESULTS: SCFA transport was significantly reduced in the presence of branched and unbranched SCFAs and several bromo, chloro and mercapto analogues as well as nicotinic acid and L lactate. In contrast, known inhibitors of monocarboxylate transporter proteins like stilbene derivatives, phloretin and 2-cyano-4-hydroxycinnamate did not inhibit bicarbonate-gradient stimulated butyrate transport. Kinetic analysis of increasing substrate concentrations of 3-mercaptopropionate, L-lactate and nicotinic acid showed saturation kinetics with apparent K(i) of 6.1, 18.3 and 14.7 mmol/l, respectively. CONCLUSIONS: The data not only confirm earlier results that absorption of SCFAs in apical membranes of the rat distal colon is mediated by a relatively low affinity/high capacity SCFA(-)/HCO(-)(3) exchange mechanism, but also indicate that although this anion transporter shares some functional similarities, is not identical with the recently cloned MCT isoforms. PMID- 11025364 TI - Local and systemic zymogen activation in human acute pancreatitis. AB - BACKGROUND: Activation of trypsinogen and phospholipase A(2) is an early event in pancreatic inflammation, but little is known about zymogen activation and the severity of human pancreatitis. METHODS: Using a new fluoroimmunoassay we measured trypsinogen activation peptide (TAP) and phospholipase A(2) activation peptide (PROP) in plasma and ascites in 25 patients with acute pancreatitis. TAP, PROP, Pro-PROP and pancreatic PLA(2)-I were measured in plasma for 14 days and in pancreatic necroses, ascitic fluid and pleural effusions. RESULTS: All 16 patients with severe acute pancreatitis (SAP) had pancreatic necrosis, 10 developed systemic complications like sepsis, pulmonary or renal failure, 6 had infected necrosis, and 4 died. All 9 patients with mild pancreatitis (MAP) survived. Plasma TAP on admission was higher in patients with SAP than in those with MAP and increased in infected necroses. It did not correlate with systemic complications. Systemic PROP was not increased in complicated courses but was significantly higher in patients with MAP than in those with SAP on admission. Pro-PROP was higher in patients with SAP than in those with MAP but was not correlated with systemic complications. Plasma pancreatic PLA(2)-I was increased but not different in patients with SAP and those with MAP. In patients with pancreatic necrosis, TAP and PROP were highest, while in those with post-acute pancreatic abscess, only PROP and Pro-PROP were high. In patients with pleural effusion, TAP was low and PROP/ Pro-PROP were high. CONCLUSION: Trypsinogen and PLA(2)-I activation are early events in acute pancreatitis and the activation peptides can be detected in plasma. In the pancreas, trypsinogen activation is accompanied by PLA(2)-I activation in patients with pancreatic necrosis. However, in our study, organ complications in SAP patients was not associated with increased plasma PROP. PMID- 11025363 TI - Induction of artificial fat maldigestion by tetrahydrolipstatin assessed by the (13)C-hiolein breath test in healthy volunteers. A double-blind controlled pilot study. AB - BACKGROUND: (13)C-hiolein is a purified algal triglyceride which is synthesized with 98% of all carbon atoms of the molecule uniformly (13)C-labeled. Orlistat is the new name for tetrahydrolipstatin (THL), which is a specific and potent inhibitor of gastrointestinal lipases which induces artificial fat malabsorption in humans. The aim of the present study was to use the (13)C-hiolein breath test (HBT) to assess THL-induced fat malabsorption in healthy volunteers. METHODS: Eight healthy volunteers of normal body weight underwent 2 study periods of 4 days of diet with or without THL 120 mg t.i.d. On day 5, a HBT was performed. (13)CO(2) recovery in breath samples was measured over 24 h by isotope ratio mass spectrometry. RESULTS: The peak (13)CO(2) excretion occurred only after 5 h in both treatment groups with little difference during the first 4 h between the groups. THL potently reduced fat digestion and absorption with the most pronounced effect observed after 8 h: 1.1+/-0.2 vs. 2.3+/-0.3% dose in control experiments (p<0.05). The 24-hour cumulative (13)CO(2) excretion was also significantly reduced by THL: 14.9+/-2.2 vs. 28.4+/-4.1% dose in control experiments (p<0.05). In 6 of the 8 subjects, the cumulative (13)CO(2) excretion was lower with THL 120 mg t.i.d. than with placebo. CONCLUSION: The HBT well reflects THL-induced artificial fat maldigestion in healthy volunteers. PMID- 11025365 TI - Low frequency of p16(INK4a) alterations in insulinomas. AB - BACKGROUND/AIMS: The molecular mechanisms contributing to the tumorigenesis of insulinomas are poorly understood. Disruption of the cell cycle due to inactivation of the p16(INK4a) tumor-suppressor gene was identified in a variety of human tumors, including gastrinomas and nonfunctioning endocrine pancreatic carcinomas. In this study the role of p16(INK4a) in the tumorigenesis of insulinomas was evaluated. METHODS: Seventeen insulinomas (14 benign, 3 malignant) were analyzed for genetic alterations in the p16(INK4a) tumor suppressor gene by SSCP, PCR-based deletion and methylation-specific assays. p16 expression was determined by immunohistochemistry. RESULTS: One malignant insulinoma showed a homozygous deletion of p16(INK4a) and another two benign insulinomas revealed aberrant methylation of the p16(INK4a) promoter region. All three tumors lacked p16 expression according to immunohistochemistry. None of the insulinomas carried intragenic p16(INK4a) mutations. In total, 17% of insulinomas had p16(INK4a) alterations. CONCLUSIONS: The p16(INK4a) tumor-suppressor gene contributes to tumorigenesis in only a small subset of insulinomas. PMID- 11025366 TI - Involvement of CCK in the paraventricular nucleus of the hypothalamus in the CNS regulation of colonic motility. AB - The effects of cholecystokinin octapeptide (CCK(8)), the CCK-A receptor antagonist, MK-329, and the CCK-B receptor antagonist, L-365, 260, microinfused into the paraventricular nucleus of hypothalamus (PVN) on colonic motor function was investigated in awake rats, chronically implanted with a microinjection cannula into the PVN and a catheter into the proximal colon. In fasted rats, bilateral microinfusion of CCK(8) at doses of 1.5 and 3.0 microg/rat into the PVN stimulated colonic transit, as shown by a significant increase in the geometric center by 47 and 54%, respectively. This effect of CCK(8) was site-specific to the PVN, since microinjection of the peptide into sites outside of but adjacent to PVN had no effect. In non-fasted rats, L-365,260 bilaterally microinjected into the PVN at a dose of 1.5 microg/rat inhibited propulsive colonic motor function; colonic transit time significantly increased by 73% in comparison to the control condition. Microinfusion of the CCK-A antagonist into in the PVN did not affect colonic transit. These results show that the PVN is a responsive site for the central CCK(8)-induced modulation of colonic motility. The data suggest, that endogenous CCK in the paraventricular nucleus of the hypothalamus unfolds a stimulatory effect on colonic transit through action on CCK-B receptors. PMID- 11025367 TI - Plasma cholecystokinin, plasma peptide YY and gallbladder motility in patients with slow transit constipation: effect of intestinal stimulation. AB - BACKGROUND/AIM: Because cholecystokinin and peptide YY are gut hormones with potent effects on gastrointestinal motility, we determined whether abnormalities of cholecystokinin and peptide YY exist in slow transit constipation. METHODS: Plasma concentrations of these hormones before, during and after intraduodenal infusion of a liquid meal in 21 patients with slow transit constipation were compared with the results in 8 healthy controls. RESULTS: Fasting levels of plasma cholecystokinin (3.1+/-0.2 vs. 2.4+/-0.2 pM; p = 0.02) were higher in patients. Basal plasma peptide YY (11.4+/-1.4 vs. 8.9+/-0.7 pM; p = 0.1) tended to be higher in patients. After the meal (60-90 min), incremental cholecystokinin (p<0.05), but not peptide YY, was significantly higher in patients. During intraduodenal infusion of the meal (0-60 min), incremental plasma cholecystokinin (251+/-20 pM.min) and peptide YY (1,146+/-186 pM. min) in patients were almost similar to control values (262+/-22 and 901+/-166 pM. min). Gallbladder volumes before, during and after the meal were not different between the 2 groups. Gastric emptying of a solid meal was delayed in the majority of patients (12 of 18). Abnormalities of plasma cholecystokinin were observed only in patients with delayed gastric emptying. CONCLUSION: Plasma levels of cholecystokinin are elevated in the fasting state and decrease more slowly after stimulation, but maximum release in response to intestinal nutrients is not altered in patients with slow transit constipation. The abnormality seems to be confined to a subgroup of patients with delayed gastric emptying. PMID- 11025368 TI - [1-(13)C]Galactose breath test for quantitative measurement of liver function in a short period. AB - BACKGROUND: Using a rat model of hepatectomy, we investigated whether the severity of hepatopathy could be quantitatively measured from changes in expiratory (13)CO(2) levels after intravenous administration of [1 (13)C]galactose. MATERIALS AND METHODS: Under nembutal anesthesia, 100 mg/kg [1 (13)C]galactose was administered to rats via the femoral vein, and expiratory (13)CO(2) levels were measured for 60 min. Then, 30, 70 or 90% hepatectomy was performed. In the control group, simple laparotomy was performed. Breath test was conducted 20 min after laparotomy. We examined the correlation of total (13)CO(2) output (S) or single point (13)CO(2) level (SP) every 5 min until 30 min, and at 45 and 60 min with liver weight/body weight (LW/BW) (%). RESULTS: In the control group, the breath test graph reached a plateau level, but in all groups undergoing hepatectomy a plateau level was not reached during measurement. The correlation coefficient between S(30) after [1-(13)C]galactose administration and LW/BW was 0.889 (p< 0.0001). The correlation coefficient between SP(25) after [1 (13)C]galactose administration and LW/BW was highest, 0.923 (p< 0.0001). CONCLUSION: In the breath test with intravenously administered [1 (13)C]galactose, hepatopathy could be evaluated by measuring S(30) and hepatopathy could be more accurately quantitatively evaluated by measuring SP(25) over a short period. PMID- 11025369 TI - Clastogenic factors as biomarkers of oxidative stress in chronic hepatitis C. AB - BACKGROUND/AIMS: Clastogenic factors (CFs) are composed of lipid peroxidation products, cytokines and other oxidants with chromosome-damaging properties. They are regularly observed after radiation exposure and in chronic inflammatory diseases, where they are supposed to be risk factors for carcinogenesis. It appeared of interest to investigate their presence in the plasma of patients with chronic hepatitis C. METHODS: CFs are detected by chromosomal breakage studies. They were compared to malondialdehyde (MDA), total plasma thiols (t-SH), alanine aminotransferase (ALT), viral load and histological data. RESULTS: CFs were increased in 19 of 20 patients, 16 had increased MDA levels and 15 had decreased t-SH levels. Mean values were significantly different from the 20 controls (p<0.001). After the first 3 months of interferon treatment, all three markers showed significant improvement, but were not completely normalized. There was a positive correlation between CFs and necroinflammatory activity (p<0.03), while MDA was correlated with fibrosis (p<0.03). Viral load was correlated with necrosis and inflammation (p<0.05). CONCLUSION: The presence of CFs in chronic hepatitis C confirms the occurrence of oxidative stress in this disease and could be useful in clinical trials for testing antioxidants. The CF test is a sensitive assay for the detection of oxidative stress and correlates with necroinflammatory activity. PMID- 11025370 TI - Multimicrobial sepsis including Clostridium perfringens after chemoembolization of a single liver metastasis from common bile duct cancer. AB - A 65-year-old woman underwent resection of a distal common bile duct carcinoma (Whipple's procedure). Twelve months later a single hepatic metastasis was detected and a chemoembolization was performed. Immediately after chemoembolization the patient developed a multimicrobial sepsis including Clostridium perfringens. CT scans depicted pathognomonic signs of gas-containing abscess in the necrotic liver metastasis. She was subsequently treated with broad spectrum antibiotics, abscess drainage and hyperbaric oxygen therapy. We conclude that antibiotic prophylaxis is recommendable for chemoembolization of liver metastasis in patients with risk factors like intestinal biliary reflux (bilioenteric anastomosis or papillotomy and biliary stenting) and bile duct cancer, otherwise severe sepsis including clostridium bacteremia may occur. PMID- 11025371 TI - Multiple tumors and a novel E2F-4 mutation. A case report. AB - Defects in the DNA mismatch-repair are known to cause microsatellite instability (MSI) in hereditary nonpolyposis colorectal cancer (HNPCC) as well as sporadic colorectal cancer (CRC). We previously reported that the E2F-4 gene, which encodes an important transcription factor in cell cycle control, had frequent tumor-specific mutations at the trinucleotide coding region microsatellite (CAG)n in a subset of human sporadic CRC with MSI. We report a 65-year-old man with triple tumors in the abdomen, including colon cancer, stomach cancer, and lipoma of the retroperitoneum, with the analysis of E2F-4 mutation. We report the first case of colon cancer with a homozygous E2F-4 mutation along with a detailed analysis of other cancer related genes as well as a prognosis. PMID- 11025372 TI - Intermittent androgen blockade in prostate cancer: rationale and clinical experience. AB - Cancer of the prostate continues to be one of the most common malignancies in men in Europe, with a large number of patients presenting with advanced disease. The current standard treatment for metastatic cancer of the prostate, permanent androgen withdrawal, is palliative. Patients treated with permanent androgen blockade usually relapse and die secondary to prostate cancer's ability to progress to an androgen-independent state of growth. Based on experimental and preclinical studies, intermittent androgen blockade appears to be a potential alternative to permanent androgen blockade. Through the cycling of reversible androgen suppression, there appears to be recovery of apoptosis and subsequent slower progression to an androgen-independent state. In this paper experimental and preclinical studies concerning intermittent androgen blockade are reviewed. At present several prospective randomized trials are under way to test intermittent androgen blockade as an alternative treatment in various stages of cancer of the prostate. However, until the results of these trials are available, this approach remains experimental. PMID- 11025373 TI - Radical prostatectomy can provide a cure for well-selected clinical stage T3 prostate cancer. AB - OBJECTIVE: Radical prostatectomy is commonly believed not to achieve the eradication of locally advanced disease. This retrospective study aimed to elucidate the role of radical prostatectomy in this condition. METHODS: A retrospective study of 158 patients surgically treated for clinical stage T3N0M0 prostate cancer was undertaken. Thirty patients had postoperative hormonal treatment, rendering prostate-specific antigen (PSA) follow-up unreliable, and were considered to be progressive at 1 month. Eighteen other patients received postoperative radiotherapy. One hundred and ten patients had radical prostatectomy only. PSA-relapse-free survival was analyzed. The mean follow-up time was 30 months. RESULTS: Seventy-nine percent of the resected specimens were pathologically T3 (pT3), and about 25% were pT3c. Thirteen percent were pT2 and 8% were pT4. Ninety-five specimens (60%) had positive surgical margins. There was poor accordance between the biopsy Gleason score and that of the specimen. A multivariate analysis showed that seminal vesicle and nodal invasion, margin status and a PSA level above 10 ng/ml were independent prognostic factors. In 47 cT3a patients with PSA <10 ng/ml, the PSA-free survival rate exceeded 70% at 24 months and the 5-year estimated PSA-free survival rate was more than 60%. CONCLUSIONS: Radical prostatectomy has a place in the treatment of clinical stage T3 prostate cancer patients with a PSA value lower than 10 ng/ml. There is a need to definitively rule out nodal or seminal vesicle invasion in order to select those patients that can benefit from surgery. PMID- 11025374 TI - Comparisons of the various combinations of free, complexed, and total prostate specific antigen for the detection of prostate cancer. AB - OBJECTIVES: We compared the ability of three prostate-specific antigen (PSA) ratios - free-to- total PSA ratio (fPSA/tPSA), free-to-complexed PSA ratio (fPSA/cPSA), and complexed-to-total PSA ratio (cPSA/tPSA) - to distinguish prostate cancer from benign prostatic hyperplasia (BPH). METHODS: We tested 258 consecutive patients who underwent transrectal ultrasound-guided prostate needle biopsy because of an abnormal digital rectal examination or a Tandem-R PSA of >4.1 ng/ml. Free PSA (fPSA) and total PSA (tPSA) were measured by Tandem-R assay. alpha(1)-Antichymotrypsin-complexed PSA (cPSA) was measured by Markit-M PSA-ACT assay. RESULTS: Of the 258 patients, 204 had BPH, and 54 had prostate cancer. The specificity at 96% sensitivity for fPSA/tPSA, fPSA/cPSA, and cPSA/tPSA was 23, 25, and 33%, respectively. Of 162 patients with tPSA between 4.1 and 10.0 ng/ml, 132 had BPH and 30 had prostate cancer. The specificity at 96% sensitivity for f/tPSA, f/cPSA and c/tPSA was 32, 44, and 41%, respectively. There was no significant difference in the area under the receiver-operating characteristic curves among fPSA/tPSA, fPSA/cPSA, and cPSA/tPSA in the overall PSA range or in tPSA between 4.1 and 10.0 ng/ml. CONCLUSION: fPSA/tPSA, fPSA/cPSA, and cPSA/tPSA did not differ in their ability to distinguish prostate cancer from BPH. PMID- 11025375 TI - Incidence, mortality and long-term survival from prostate cancer in Umbria, Italy, 1978-1994. AB - OBJECTIVES AND METHODS: Incidence, mortality and long-term survival from prostate cancer were examined in the Umbria region of Italy, for the period 1978-1994. Incidence rates were derived from an ad hoc survey carried out over the period 1978-1982 and from 1994 cancer registry records. The mortality over the period derived from data of the official publications and the survival rates, at 15 years, were calculated starting from the ad hoc survey incident cases. RESULTS: In the Umbria region, over the period 1978-1994, crude incidence rates from prostate cancer increased from 31.2 to 81. 9 per 100,000 and mortality rates from 22.7 to 31.9 per 100,000. The rates were from 30.5 to 61.2 for age-adjusted incidence while standardized mortality remained constant (from 22.6 to 22.7 per 100, 000). Survival in Umbria, compared with rates from other European Cancer Registries, is low both at 5- and 10-year follow-up. CONCLUSION: The great incidence increase observed over the study period could depend on a lesser completeness achieved by the 1978-1982 ad hoc survey with respect to the 1994 cancer registry data and/or from the screening campaign carried out in a large part of the region in 1994. Different elements support these hypotheses. However the above hypotheses can be verified over the next years when further incidence and survival data from the cancer registry will be available. PMID- 11025376 TI - Perisphincteric injection of botulinum toxin type A. A treatment option for patients with chronic prostatic pain? AB - BACKGROUND: Chronic prostatic pain is still a diagnostic and therapeutic problem. The clinical observation that prostatic and pelvic pain is accompanied by motoric and sensoric disorders of the pelvic floor muscles led to the hypothesis that prostatic pain roots in a changed processing of afferent and efferent information with the central nervous system (CNS). METHODS: Neuro-urological work-up of 11 male patients with chronic prostatic pain was completed. This included a clinical evaluation of pelvic floor function, urodynamic investigation of bladder and urethra function and a cystoscopy to exclude morphological aberrations. A transurethral perisphincteric injection of 200 units botulinum toxin type A (BTX) was followed by a 2- to 4-week visit to evaluate their influence on the neuro urological symptomatology. RESULTS: All chronic prostatic pain patients suffered from a pathological pelvic floor tenderness, an inability of sufficient conscious pelvic floor control, a urethral hypersensitivity/hyperalgesia and a urethral muscle hyperactivity. Basic parameters of bladder function (capacity, sensitivity, compliance) were normal. The BTX injection was followed by a pelvic floor muscle weakening and a relief of prostatic pain and urethral hypersensitivity/hyperalgesia. A botulinum-related decrease of the functional urethral length, the urethral sphincter closure pressure, the postvoid residual volume and an increase of the peak and average uroflow were objectivated. CONCLUSION: A weakening of the urethral sphincter muscle via blocking acetylcholine release by BTX injection is followed by pain relief and symptom improvement. It can therefore be concluded that a barrage of nociceptive information from the dysfunctional pelvic floor overflood the CNS and induce a changed CNS processing. Interrupting the efferent branch of the disturbed central circle is one opportunity to treat chronic prostatic pain. PMID- 11025378 TI - Experience in the surgical treatment of Fraley's syndrome. AB - OBJECTIVE: To report our 20-year experience with nephron-sparing surgery in the treatment of Fraley's syndrome. MATERIALS AND METHODS: From September 1976 to July 1996, 6 women, 18-43 years old, underwent surgical treatment. All patients had been suffering debilitating right flank pain for at least 6 months before the operation. Diagnosis was in call cases made with the use of intravenous pyelography and renal arteriography, which showed a vascular impression on the superior infundibulum with secondary dilatation of the upper pole calyx. This was localized only on the right side in 5 cases, while in 1 it was bilateral though more severe on the right. Ipsilateral nephroptosis was observed in 2 of the patients. In 2 cases in whom the superior infundibulum was sufficiently long, an infundibulo-infundibulostomy with everted flaps was performed. In a case in whom a short infundibulum was compressed between a venous and an arterial branch, Fraley's infundibulopyelostomy was carried out. In 1 patient in whom the infundibulum was compressed by the anterior-superior segmental artery, a Heineke Mikulicz-type infundibulorrhaphy was combined to vasopexy. In the remaining 2 cases, the infundibulum was not cut: in one case, a simple vasopexy of two vascular branches was carried out, while in the other, a minor arterial branch was ligated and divided. In 4 patients, nephropexy was also performed. RESULTS: No significant intra- or postoperative complications were observed. Follow-up averaged 102.5 months. Pain relief was complete in 5 cases, in whom disappearance of the vascular impression was also radiologically demonstrated. Only 1 patient, who underwent infundibulorrhaphy and vasopexy, experienced occasional flank pain and urinary infection after the operation, with just a slight improvement in the excretory urogram. CONCLUSIONS: Surgical treatment of Fraley's syndrome is indicated only in symptomatic and/or complicated cases; in relation to the type of obstruction and the anatomy of the intrarenal structures, whether excretory or vascular, several effective nephron-sparing techniques can be selected. PMID- 11025377 TI - Double-blind trial of the efficacy and tolerability of doxazosin in the gastrointestinal therapeutic system, doxazosin standard, and placebo in patients with benign prostatic hyperplasia. AB - BACKGROUND: The alpha(1)-blocker doxazosin mesylate is an established efficacious and welltolerated treatment for benign prostatic hyperplasia (PBH). However, its clinical utility can be limited by the need for multiple titration steps, starting at an initial dose of 1 mg, increased up to 8 mg once daily, to achieve optimal therapeutic response. A new controlled-release gastrointestinal therapeutic system (GITS) formulation of doxazosin mesylate enhances the pharmacokinetic profile and drug delivery rate, reducing the plasma doxazosin mesylate peak-to-trough ratio and minimizing the need for titration. OBJECTIVE: A study was conducted to assess the effects of doxazosin GITS 4 or 8 mg once daily, doxazosin standard 1 mg to 8 mg once daily, and placebo, in 795 men with BPH. This randomized, double-blind, multicenter Scandinavian study included a 2-week washout period, 2-week single-blind placebo run-in phase, and 13-week double blind treatment phase. Doxazosin GITS was initiated at 4 mg once daily and titrated to 8 mg once daily after 7 weeks, if indicated, and doxazosin standard was initiated at 1 mg once daily, titrated to 2 mg after 1 week, to 4 mg at 3 weeks, and to 8 mg at 7 weeks if indicated, to achieve symptom control. The primary outcome measures were mean changes from baseline to the final visit for International Prostate Symptom Score (I-PSS) and maximum urinary flow rate adjusted for baseline values. RESULTS: Both doxazosin GITS and doxazosin standard significantly improved the symptoms of BPH, as evidenced by least-squares mean reductions in total I-PSS of -8.0+/-0.3 and -8.4+/-0.3 from baseline, respectively, compared with a reduction of -6.0+/-0.4 in patients on placebo. Doxazosin GITS and doxazosin standard produced clinically comparable improvements in maximum urinary flow rates, with a greater improvement observed earlier following treatment with doxazosin GITS than with doxazosin standard. Both active treatments produced significantly greater increases in maximum urinary flow rate compared with placebo. Nearly half of the patients on doxazosin GITS achieved symptom relief at the 4-mg starting dose. A similar number of patients in both doxazosin groups were titrated to the maximum dose of 8 mg for both formulations. The overall incidence of adverse events was similar among patients treated with doxazosin GITS and placebo, and slightly higher in those on doxazosin standard. There was no apparent difference in the type of adverse events reported for the two formulations of doxazosin, although most adverse events were reported at a lower frequency with doxazosin GITS. CONCLUSIONS: Doxazosin GITS is significantly more effective than placebo in reducing the clinical symptoms of BPH and improving maximum urinary flow rate, and as effective as doxazosin standard. A therapeutic effect equivalent to that of doxazosin standard was achieved with doxazosin GITS with fewer titration steps, in a manner that appeared to be better tolerated. Because treatment with doxazosin GITS starts with an effective dose for many patients, it is likely that this clinical profile will result in the need for fewer patient visits than with doxazosin standard therapy. PMID- 11025379 TI - Urodynamic evaluation and management prior to renal transplantation. AB - OBJECTIVES: To evaluate the indications for urodynamics studies (UDS) previous to renal transplantation (RT) and the results of bladder cycling (BC), performed in selected cases. METHODS: Among 475 RT, 33 patients (6.9%) required UDS according our protocol. Indications were: (1) low urinary tract symptoms (LUTS); (2) defunctionalized bladder (DB), and (3) complex urologic history (reflux, neurogenic bladder, urethral valves, etc.). DB was defined as a total urinary output of <300 ml/24 h. Patients with DB underwent BC through a suprapubic catheter, undergoing UDS after BC. The rate and type of urodynamic anomalies were compared among the 3 groups. Cystometric capacity (CysCap) and compliance were evaluated according to the etiology of renal failure, diuresis, and number of previous bladder surgeries. RESULTS: Fifteen patients (45%) showed abnormal UDS, Increased pressure at cystometry was the most frequent finding (46%). The rate of abnormal UDS among the 3 groups was not statistically significant (66.6% in DB, 42.8% in LUTS, and 35% in those with complex urologic history). Compliance and CysCap were significantly lower in the patients with DB, showing a significant correlation with diuresis. These parameters did not correlate with the number of previous bladder surgeries. BC resulted in normal UDS in 3 patients (33%) and a change in diagnosis in 2 patients (22%). CONCLUSION: Our indications gave a good diagnostic yield. Patients with DB represent a group with different urodynamic findings. BC provides a more precise diagnosis or normalization of UDS in some cases. PMID- 11025380 TI - Urinary BTA-stat, BTA-trak and NMP22 in surveillance after TUR of recurrent superficial transitional cell carcinoma of the bladder. AB - OBJECTIVES: To evaluate NMP22, BTA-Stat and BTA-Trak tests in monitoring recurrent transitional cell carcinoma of the bladder. METHODS: The tests were performed in 179 selected patients being followed up for recurrent superficial bladder tumors: 55 patients had bladder recurrence and 124 patients were recurrence free. The NMP22 test was obtained in all patients; BTA-Stat and BTA Trak in the last 96 and 74 patients, respectively. Sixty-four patients (51.6%) were undergoing adjuvant intravesical chemotherapy. RESULTS: Sensitivity was 74, 57 and 62% and specificity was 55, 62 and 79% for NMP22, BTA-Stat and BTA-Trak, respectively. A high percentage of patients submitted to intravesical chemotherapy had false-positive tests. Positive predictive values of the NMP22, BTA-Stat and BTA-Trak tests were 42.2, 40 and 45.4%, and negative predictive values were 82.9, 76.9 and 88.4%, respectively. CONCLUSIONS: NMP22, BTA-Stat and BTA-Trak tests cannot replace cystoscopy and cannot be adopted as routine tools in surveillance after TUR in patients with superficial bladder cancer. PMID- 11025381 TI - Genetic analysis of Tp53 from urine sediment as a tool for diagnosing recurrence and residual of bladder carcinoma. AB - OBJECTIVES: Longitudinal study of Tp53 mutation in urine sediments of 26 patients with mutated primary transitional cell carcinoma (TCC) of the urinary bladder at different time periods after transurethral resection of the bladder (TURB), i.e. before and after the first TURB, prior to the control resection and before treatment of a recurrence. METHODS: DNA of the critical Tp53 exons 5-8 was anaylzed by temperature gradients (TGGE) and sequence. RESULTS: (1) In 11 of 12 patients (91.7%) mutation reoccurred with the detection of recurrence of the disease. The mutation frequency in patients without any recurrence was 1 in 8 (12.5%) after a follow-up period of 4-16 months. (2) In 7 of 10 patients, the mutation was no longer detectable in the urine sediment after TURB. (3) The mutation frequency at the control resection 6 weeks after the first TURB was 5 in 7 (71.4%) in patients found to have residual and 1 in 7 (14.2%) in the tumor-free patients. (4) In 9 of 10 samples identical mutations were found by sequence in the recurrent tumor. These results show a significant correlation between the detection of a Tp53 mutation in the urine sediments and tumor recurrence or residual. CONCLUSIONS: (1) Tp53 mutations in the urine sediment could be a useful indicator of tumor recurrence or tumor residual in patients ( approximately 40%) with primary mutated bladder cancer tissue. (2) These results support the monoclonal seeding theory. (3) The finding of identical mutations at different times indicate that the tumor was never totally removed. PMID- 11025382 TI - Is suprapubic cystostomy an optimal urinary management in high quadriplegics?. A comparative study of suprapubic cystostomy and clean intermittent catheterization. AB - INTRODUCTION: Long-term outcome of spinal cord injury (SCI) patients was compared between those managed by suprapubic cystostomy (SPC) and clean intermittent catheterization (CIC) with particular emphasis on an incidence of urinary tract complications and patients perception for urinary management. MATERIALS AND METHODS: The study comprised 61 SCI patients; 34 patients managed with SPC (group A), while 27 with CIC (group B). After stabilization of their condition, all were followed annually on an outpatient basis with clinical history, urinalysis, urinary imaging and renal function studies. Mean follow-up periods were 8.6 and 9.9 years for groups A and B, respectively. Between groups, a comparative study was performed on the incidence of urinary complications such as renal dysfunction, hydronephrosis, vesicoureteral reflux, symptomatic genitourinary infection and urinary stone. Satisfaction with urinary management was also estimated using the questionnaires during follow-up. RESULTS: Renal dysfunction, hydronephrosis and vesicoureteral reflux were not found in either group. Symptomatic genitourinary infection was seen in 4 (12%) of group A and 7 (26%) of group B, respectively. The incidence of renal stone was 3 (9%) in group A and 1 (4%) in group B. A significant difference was not found between two groups in these urinary complications. On the contrary, bladder stone was seen more frequently in group A (65%) than in group B (30%) with a significant difference (p<0.001). The degrees of incontinence, bother score of daily activities, and overall satisfaction showed no significant difference between the two groups. CONCLUSION: Except for bladder stones, SPC is a valuable option of urinary management for quadriplegic patients, the results of which were comparable to paraplegic SCI patients managed with CIC. PMID- 11025383 TI - Sacral neuromodulation for women with Fowler's syndrome. AB - Neuromodulation of the sacral nerves has been found to be an effective therapy for a variety of lower urinary tract dysfunctions. The reported success rate for the period of trial stimulation (peripheral nerve evaluation test) prior to permanent implantation of a sacral nerve stimulator is variable, but generally reported to be in the region of 30-50%. We present here the results of the peripheral nerve evaluation test in 38 patients with urinary retention. 34 of the 38 had been found to have an abnormality of their striated urethral sphincter on electromyography using a concentric needle electrode, i.e., they had the disorder which was described by Fowler and coworkers in 1988. The overall success rate in this group was 68%. We believe that our relatively high success rate is due to sacral neuromodulation working via a mechanism which involves the urethral sphincter, an abnormality which had been demonstrated in 89% of these patients. Twelve of the patients subsequently underwent permanent implantation of a sacral nerve stimulator, and all of them have experienced a return of voiding. However, in 2 patients, there is a persisting need for self-catheterization. There is, however, a high reoperation rate. PMID- 11025384 TI - Desensitization of bladder sensory fibers by intravesical resiniferatoxin, a capsaicin analog: long-term results for the treatment of detrusor hyperreflexia. AB - OBJECTIVES: To assess the pungency and the desensitizing effects of intravesical resiniferatoxin, an ultrapotent capsaicin analog, in patients with detrusor hyperreflexia. METHODS: Fourteen patients with detrusor hyperreflexia were instilled during 30 min, without any form of local anesthesia, with 100 ml (or the bladder capacity if lower than that volume) of 50 or 100 nM resiniferatoxin solutions in 10% alcohol in saline. Patients were evaluated by voiding chart and urodynamic tests (volume to first contraction, maximal cystometric capacity, maximal detrusor pressure, icewater test) at 14, 30, 60, and 90 days and every 3 months thereafter RESULTS: Resiniferatoxin instillation did not evoke pain or temporary worsening of urinary symptoms. Treatment improved or abolished incontinence in 9 out of 12 incontinent patients. Mean urinary frequency decreased from 14.2+/-6.4 to 10.3+/-3.2 at 3 months (p = 0.01). At this time point, mean maximal cystometric capacity increased from 182.3+/-119.8 to 330.0+/ 201.6 ml (p = 0.01) and the ice water test, positive in 13 cases, became negative in 8 of them. Maximal detrusor pressure was not modified by the treatment. The effect was long-lasting, reaching 12 months in 7 cases. CONCLUSIONS: The absence of irritative symptoms during bladder instillation of resiniferatoxin and the rapid onset of desensitization make this vanilloid superior to capsaicin for the treatment of detrusor hyperreflexia. PMID- 11025385 TI - Bladder neck closure for treating pediatric incontinence. AB - AIMS OF STUDY: In order to evaluate the effects of bladder neck closure (BNC) for treatment of pediatric incontinence, on the quality of life of those children, we reviewed the files of 17 children who underwent this procedure during the last 5 years. Information on previous surgery before BNC, continence and complications after BNC and patient satisfaction are gathered. MATERIAL AND METHODS: During the study period, 17 children (9 male, 8 female) underwent BNC with continent diversion. Ten children (5 male, 5 female) suffered neurogenic incontinence in meningomyelocele, 5 children (4 male, 1 female) had bladder exstrophy, 1 girl suffered iatrogenic incontinence after treatment of an ectopic ureterocele and 1 girl suffered structural incontinence after pelvic fracture. Previous surgery for incontinence was done in 12 children with 36 procedures. Primary BNC was done in 5 children. For continent diversion the appendix was used in 13, the ureter in 2, a Monti procedure in 1, and an ileal valve in 1. In 9 children, ileal bladder augmentation was performed at the time of BNC. Four children were augmented before. Mean age at time of operation was 13.5 years. Mean follow-up is 35 months. RESULTS: After BNC, all patients were completely dry. One girl suffered some stomal incontinence during the night. This disappeared after recent bladder augmentation. Patient satisfaction is extremely high after surgery. All patients feel happy with their stoma and do not regret the choice they made. As for complications, urinary tract infections were seen in 9 patients. Stomal complications were seen in 8 patients. Three patients had some difficulty with catheterization, which could be solved with dilatation. Appendiceal polyps, which could be removed under local anesthesia, were seen in 3 and stomal stenosis which needed re-intervention in 2. In 1 patient, three revisions of the stoma were done, in the other patient, two surgical corrections. Out of 8 patients showing stomal complications, only 2 needed revision under anesthesia. All complications are seen in the first 6 months after the continent diversion. CONCLUSIONS: We present a series of patients who underwent BNC for treatment of incontinence. BNC is the ultimate bladder neck reconstruction. However, regarding the high success rate, the low complication rate and the high patient satisfaction, we must consider BNC as an important procedure for the reconstructive surgeon. If reconstruction fails, closure must be considered. Compared to most other procedures for bladder neck reconstruction, BNC gives the highest continence rate. Good patient compliance and strict medical follow-up of the upper tract is mandatory in these patients. PMID- 11025386 TI - Relevance of infection in children with urachal cysts. AB - OBJECTIVE: To assess the role of infection in the management of children with urachal cysts. METHODS: A retrospective study on 10 children with urachal cysts operated on over an 11-year period (from 1987 to 1998) was performed. Uncomplicated urachal cysts were found in 2 children who underwent primary cyst removal. The remaining 8 were admitted with severe sepsis due to the presence of a urachal abscess; they were managed by a staged approach including percutaneous drainage and delayed cyst removal. The diagnosis of urachal cyst was readily made by ultrasound in all the 10 patients (100%). In 1 patient with urachal abscess, computed tomography provided additional information. RESULTS: The postoperative course was uneventful in 9 of 10 children (90%). A 5-year-old female patient developed peritonitis following urachal abscess rupture into the peritoneal cavity, which resulted in additional surgery and prolonged hospitalization. CONCLUSIONS: (1) Ultrasound is an excellent diagnostic tool for patients with urachal cysts. (2) A renal screening ultrasound must be included in the preoperative work-up. (3) A thorough urological assessment is indicated in patients with abnormal renal ultrasound of recurrent urinary infections. (4) At present, a staged surgical procedure still remains the most effective surgical option in children with urachal cyst. PMID- 11025387 TI - Hydatid cyst of the kidney. A report of 147 controlled cases. AB - OBJECTIVES: We report our experience in the management of 147 hydatid cysts of the kidney over an 11-year period. MATERIAL AND METHODS: The records of 147 patients operated for hydatid cysts of the kidney between 1985 and 1996 were reviewed in order to address patient's symptoms at presentation, radiological findings, diagnostic tests and surgical outcome. All patients were managed with open surgery. No preoperative or postoperative antiparasitic medication was used. RESULTS: Lumbar or lumbo-abdominal pain was the most frequent symptom (84%). Hydaturia was observed in 28% of the cases. Preoperative diagnosis was based upon intravenous pyelography (IVP), ultrasound and serology tests. CT scan was performed only in litigating cases (15%). In 20 cases, damage to the renal parenchyma was so extensive as to justify nephrectomy. Abstention concerned 5 involutive type V cysts. The remaining patients had an excision of the prominent part of the cyst (partial cystopericystectomy). Postoperative course was smooth in all cases. Postoperative IVP (3-6 months) showed a restituo ad integrum in 80% of the cases and residual pelvicaliceal distortions in 20%. CONCLUSIONS: Diagnosis of hydatid cyst of the kidney is based mainly on ultrasonography and IVP. Open surgery is the treatment of choice with excellent results. PMID- 11025388 TI - Growth-inhibitory effect of prostasomes on prostatic cancer cell lines in culture. AB - OBJECTIVES: Prostasomes, small corpuscular organelles derived from the prostate gland, were isolated from human seminal plasma for incubation with three human prostatic cancer cell lines: DU145, PC3, and LNCaP. The aim of this study was to establish any possible growth-inhibitory effect of prostasomes on prostatic cancer cell lines. METHODS: Prostasomes were isolated from human seminal plasma. We determined their growth-inhibitory effect on the prostatic cancer cell lines by a fluorimetric cytotoxic assay technique. The results obtained were presented as survival index, defined as a percentage of control cultures, after compensation for blank values. RESULTS: The prostasomes exhibited a dose dependent growth-inhibitory effect on the cells, cell line DU145 being the most sensitive one displaying a survival index (mean +/- SD) of 46+/-10% at 200 microg prostasomes (protein)/ml while the other two cell lines displayed survival indices of 93+/-6 and 93+/-9%. Heat treatment of the prostasomes abolished their growth-inhibitory effect on the cell lines, whereas 24-hour dialysis of the prostasomes did not affect their activity in this regard. Seminal plasma devoid of prostasomes exhibited less effect on the growth of these cancer cell lines, and heat treatment of the plasma did not influence the result. However, dialysis of the seminal plasma for 24 h abrogated its growth-inhibitory effect. CONCLUSION: The prostasomal growth-inhibitory effect on the cancer cell lines was different from that of the prostasome-free seminal plasma. The former effect was most probably of a proteinaceous nature while the latter effect was most likely due to the divalent cation composition of seminal plasma. In this regard, the zinc ion content seemed to be decisive. PMID- 11025389 TI - Aneuploidy of chromosome 9 and the tumor suppressor genes p16(INK4) and p15(INK4B) detected by in situ hybridization in locally advanced prostate cancer. AB - INTRODUCTION AND OBJECTIVES: The linked p16(INK4)/MTS1 and p15(INK4B)/MTS2 genes on chromosome 9p21 encode proteins that inhibit the cyclinD dependent kinases CDK4/6. Biallelic homozygous deletions involving this locus have been identified in a wide range of tumor cell lines, but in a lower frequency of primary tumors. As PCR based approaches analyzing for homozygous deletions could be confounded by unavoidable contributions of normal cells in microdissected tissue, we performed in situ hybridization (ISH) on primary prostate carcinomas to accurately evaluate p16 and p15 copy numbers on a cell-by-cell basis. MATERIAL AND METHODS: p16 and p15 loci were evaluated in 28 pT3N0M0 prostate cancer specimens. Of 28 patients, 15 (53%) were ascertained showing no recurrence (mean follow-up 61+/-17 months), 13 (47%) developed recurrences within 27+/-19 months. Tissues were provided for ISH analysis in a blinded fashion. Isolated DNA derived from P1 clone 1063 compromising p16 and p15 as well as a centromeric probe for chromosome 9 were used for hybridization. Signals were enumerated within 300 interphase nuclei per tumor specimen, and in 100 nuclei derived from 18 benign prostate tissues and 7 adjacent PIN regions. RESULTS: ISH detected aneuploid tumors in 12/13 (92%) patients with recurrence and in 5/15 (33%) without recurrence (p<0.0014). Whereas 3/7 PIN specimens associated with nonrecurrent PCA demonstrated euploidy, all 4/7 PIN associated with recurrent disease demonstrated the same aneuploidy for chr9 as the primary tumor. All benign tissues evaluated exhibited euploidy for chr9, p16 and p15. None of the PCA and PIN samples revealed homozygous deletions for p16(INK4)/MTS1/p15(INK4B)/MTS2; 2/28 (7.1%) PCA exhibited partial deletion for p16(INK4)/MTS1/p15(INK4B)/MTS2 and aneuploidy for chr9; both PCA derived from the recurrent group. CONCLUSIONS: Deletion of 9p21 was rare and therefore such genetic alterations may not play an important role in the pathogenesis of PCA. Analysis of the limited number of PCA examined suggest a strong association between chr9 aneuploidy and recurrenct disease. Aneuploidy in both PIN and PCA suggests that the clinical outcome of PCA might already be determined in the preinvasive PIN. PMID- 11025390 TI - One-stage hypospadias repair. PMID- 11025391 TI - Female incontinence PMID- 11025392 TI - [Dynamics of radon decay product activity in saliva following therapeutic radon exposure]. AB - Radon decay product activity was measured in saliva of 10 male patients 20-30 min after a 1-hour radon exposure in the gallery of the Gasteiner Heilstollen (radon activity 36.2 kBq/m(3), radon progeny activity 20.3 kBq/m(3)). In 1 patient showing relatively high activity (75th percentile) measurements were continued until 65 min after exposure. Patients were asked to collect about 2 ml of saliva in the mouth and produce it on a filter. After drying the filter at 300 degrees C, radon progeny activity was measured. Activity (median) at 20-30 min after leaving the treatment area was 4.5 Bq (25th percentile 1 Bq; 75th percentile 21 Bq). In the patient who underwent additional measurements the activity showed a further increase up to 29 Bq (35 min after radon exposure) before it continuously decreased to a very low activity (1-3 Bq) at 65 min after exposure. The results show that a significantly increased radon decay product activity is found in saliva after speleotherapeutic radon exposure. Maximum values were observed 35 min after radon exposure. Radon decay product activity almost disappeared after about 1 h. PMID- 11025393 TI - Personal values and medical preferences: postmaterialism, spirituality, and the use of complementary medicine. AB - BACKGROUND: The use of complementary medicine is increasing in the countries of the West. To find out the reason for this, research concentrated on the patients' demands for these methods, on their dissatisfaction with conventional medicine, and on their health conceptions. Quantitative research into the influence of attitudes and convictions in a broader sense on the use of complementary medicine are lacking, but would be of interest. QUESTIONS: This article aims to throw light on the specific question of whether materialistic or postmaterialistic values and spiritual preferences correlate with the use of unconventional medical methods. METHOD: Within the framework of the Swiss National Research Programme 34: 'Complementary Medicine', 3,077 and 2,276 Swiss residents were interviewed by telephone in 1995 and 1996, respectively, about their use of the medical system as well as about their attitudes towards materialism and spirituality. RESULTS: Hypotheses were confirmed: Attitudes and convictions influence the use of complementary medicine. Postmaterialists and interviewees who tended to agree with neoreligious statements used complementary medicine significantly more frequently than materialists and interviewees who tended to disagree with neoreligiosity or who tended towards traditional Christian values. CONCLUSIONS: Further research should concentrate on the interaction of different attitudes and convictions in order to learn more about the background of the growing trend towards complementary medicine. Another important conclusion is that the so called health market is not simply subject to supply and demand, and cannot be regulated by marketing means alone. PMID- 11025394 TI - [The efficacy of the complex medication Phyto-Hypophyson L in female, hormone related sterility. A randomized, placebo-controlled clinical double-blind study]. AB - OBJECTIVE AND DESIGN: In a prospective, randomized, placebo-controlled, double blind study, the effects of Phyto Hypophyson L (Steierl-Pharma GmbH, Herrsching, Germany), an Agnus castus-containing homeopathic preparation, were investigated in 67 women with fertility disorders. PATIENTS AND METHODS: 37 women with oligomenorrhea and 30 women with amenorrhea received 50 drops of Phyto Hypophyson L or placebo 3 times a day over 3 months or 3 cycles. OUTCOME MEASURE AND RESULTS: The outcome measure being spontaneous menstruation, improved concentration of progesterone in the luteal phase, shortening of the cycle, earlier ovulation, and pregnancy was achieved in 38 out of 67 women. It was achieved more often from women with oligomenorrhea in the Phyto Hypophyson L group compared to the placebo group (82 versus 45%, p = 0.021). However, there was no significant effect when viewing the whole group. The baby take-home rate during the therapy and 6 months after the end of the therapy showed a ratio of 6 : 2 (18.7 : 6.4%). This result was not significant. Furthermore, in the oligomenorrhea verum group we observed a significant increase of progesterone during the luteal phase compared to the oligomenorrhea placebo group. Only very few undesirable drug effects were observed. CONCLUSION: In women with sterility and oligomenorrhea, a treatment with Phyto Hypophyson L can be recommended over a period of 3-6 months. PMID- 11025395 TI - [Prosta Fink Forte capsules in the treatment of benign prostatic hyperplasia. Multicentric surveillance study in 2245 patients]. AB - OBJECTIVE AND DESIGN: Therapeutic use and safety of a pumpkin seed extract were tested in a multicentric clinical trial. PATIENTS AND METHODS: 2,245 patients suffering from benign prostate hyperplasia (stage I to II according to Alken) received for 12 weeks 1-2 capsules Prosta Fink Forte per day. Urinary symptoms were recorded by the International Prostate Symptom Score according to the American Urological Association (I-PSS), the influence on quality of life has been recorded by a quality of life questionnaire (LQ Index). RESULTS: The I-PSS decreased by 41.4%, life quality improved by 46. 1% during therapy. More than 96% of the patients reported no undesired side effects under the treatment with Prosta Fink Forte. CONCLUSION: Prosta Fink Forte facilitates an efficient improvement of BPH symptoms, especially in early stages. PMID- 11025396 TI - 1) rheuma: komplementare therapien hoch im kurs; 2) ist der Ruckenschmerz magnetisch? 3) morbus alzheimer: unkonventionelle therapeutische Wundertute; 4) homocystein: Im westen nichts neues AB - Wahrend in akademischen Kreisen noch ausgiebig diskutiert wird, wie Naturheilkunde, Komplementarmedizin und Alternativmedizin begrifflich fassbar gemacht, sprich definiert werden konnen, hat sich langst ein Parallelmarkt entwickelt, auf dem seriose und obskure Anbieter eine breite Palette von vermeintlich hilfreichen diagnostischen Massnahmen und Therapien feilbieten. Ohne kassenarztliche Vereinigung, ohne wissenschaftliche Fachgesellschaften, ohne amtlichen Segen. Der Markt boomt, wie das Beispiel rheumatische Erkrankungen zeigt. Das hat seine Grunde, von denen S. Hill in ihrem Kommentar einige nennt. Abstrahiert konnte man sagen, dass es offensichtlich einen grossen Bedarf in Bereichen gibt, fur die die konventionelle Medizin zu wenig zu bieten hat. Die daraus resultierende Tatsache, dass Patienten immer mehr bereit sind, zusatzlich und erganzend Leistungen einzukaufen, musste nicht weiter schrecken (und ware parteienubergreifend sogar ganz im Sinne unserer Gesundheitspolitiker), wenn man unterstellen konnte, Patienten hatten eine einigermassen verlassliche Moglichkeit, seriose Angebote von denen von Quacksalbern, Scharlatanen und miesen Geschaftemachern zu unterscheiden. Das haben Sie aber nicht. Die Beispiele in diesem Journal Club sollen aufzeigen, wie schwer es manchmal selbst dort ist, Stellung zu beziehen, wo bereits Versuche vorliegen, bestimmte Angebote zu hinterfragen. Das liegt nicht zuletzt an den ziemlich durchgangigen Defiziten, was die Forschungsqualitat in diesem Bereich anbelangt. Es scheint hochste Zeit, dass sich die akademische Medizin ihrer Verantwortung in diesem Bereich bewusst wird und sich seiner annimmt. Wer sonst sollte das tun? Copyright 2000 S. Karger GmbH, Freiburg PMID- 11025397 TI - The synthetic metalloproteinase inhibitor batimastat suppresses injury-induced phosphorylation of MAP kinase ERK1/ERK2 and phenotypic modification of arterial smooth muscle cells in vitro. AB - Smooth muscle cell (SMC) migration and proliferation are important events in the formation of intimal lesions associated with atherosclerosis and restenosis following balloon angioplasty. The extracellular matrix has important functions in modulating SMC structure and function, but less is known about the role of the matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors. The present study investigates the effects of the synthetic MMP inhibitor batimastat (BB94) on vascular SMCs. As experimental model, rat aortic smooth muscle cells in primary and secondary cultures were employed. Electron microscopy was used to investigate the effects of BB94 on the overall phenotypic properties of the cells. Induction of DNA synthesis and migration was studied by thymidine autoradiography and counting of cells moving into an injured zone. Gelatin zymography was used for the detection of BB94-mediated inhibition of injury induced MMP activity. Phosphorylation of the mitogen-activated protein kinases ERK1/ERK2, two potential mediators of the injury-induced activation of the cells, was measured by Western blotting. The results show that BB94 restrained the phenotypic modulation of vascular SMCs in primary cultures and suppressed injury induced DNA synthesis and migration. Moreover, the upregulation of ERK1/ERK2 phosphorylation in injured secondary cultures and in cells treated with bFGF was markedly reduced by BB94, whereas TIMP-2 lacked a clear effect. Our data suggest that BB94 inhibits injury-induced activation of vascular SMCs by acting on MMPs as well as other targets. PMID- 11025398 TI - Neurotrophins and neurotrophin receptors in human pulmonary arteries. AB - The localization of neurotrophins (NTs) and NT receptors was analyzed in sections of human extra- and intrapulmonary arteries by Western blot analysis and immunohistochemistry. In extrapulmonary branches of human pulmonary artery, NT and NT receptor immunoreactivity was located in the tunica intima, within endothelium, in the tunica media, within smooth muscle and in the tunica adventitia. In different sized intrapulmonary arteries, NT and NT receptor immunoreactivity was observed primarily in the tunica adventitia. A faint NT and NT receptor immunoreactivity was observed in the tunica media of large-sized branches of intrapulmonary arteries, but not within medium- or small-sized intrapulmonary vessels or in tunica intima of different sized intrapulmonary arteries. These findings suggest that NTs may have a role in the control of vascular responses in the pulmonary system acting as local paracrine or autocrine mediators. The possible relevance of the NT system in human pulmonary vasculature identified in this study is discussed. PMID- 11025399 TI - Haemopoietic origin of myofibroblasts formed in the peritoneal cavity in response to a foreign body. AB - This study utilized both in vivo and in vitro techniques to investigate whether cells of bone marrow origin can differentiate into smooth muscle-like cells (myofibroblasts) with contractile filaments and proteins. Female C57BL/6 mice expressing the Ly5.2 antigen on the surface of their haemopoietic cells had four pieces of silastic tubing (3 x 0.5 mm outer diameter) or boiled blood clot (2-3 mm diameter) placed in their peritoneal cavity. After 3, 5, 7 and 14 days (n = 4/group) the implants were removed and those that had remained free-floating were processed for light microscopy, immunohistochemistry and electron microscopy. In the first 3-5 days, rounded cells adhered to the entire surface of the tubing then flattened. These cells stained with fluoresceinated antibodies to Ly5.2 showing that they were derived from haemopoietic cells. By 14 days the cells had become elongated and multilayered in a collagen matrix, forming a thick tissue capsule around the tubing or boiled clot. They contained contractile filaments and stained with antibodies to alpha-smooth muscle actin but no longer stained for Ly5.2. A separate set of female C57BL/6 Ly5.2 mice were X-irradiated to destroy bone marrow then immediately transfused with 10(6) nucleated bone marrow cells taken from the femur and tibia of a congenic strain of male mice expressing the Ly5.1 allele. Eight of the female mice with successful engraftment (80-99%) had silastic tubing implanted in the peritoneal cavity. After 14 days, in situ hybridization with Y chromosome probe confirmed the male donor, and thus bone marrow, origin of the elongated cells that formed the capsule. In vitro studies showed that cells of the murine macrophage cell lines RAW 264.7 and J774 express alpha-smooth muscle actin after exposure to the cytokine gamma-interferon in vitro. These data show that bone marrow-derived cells can differentiate into smooth muscle-like cells and raises the possibility that blood-derived cells may contribute to the development of fibro-proliferative vascular diseases such as atherosclerosis. PMID- 11025400 TI - Gene transfer to intact mesenteric arteries by electroporation. AB - The purpose of the present study was to develop a rapid, reproducible method of nonviral gene transfer to the intact vasculature. Male Sprague-Dawley rats were anesthetized, a midline abdominal incision was made and segmental branches of the superior mesenteric artery were dissected free of surrounding mesentery. A specially designed electroporation probe was placed around the neurovascular bundle and the electroporation chamber filled with a solution containing the firefly luciferase expressing plasmid (pCMV-Lux-DTS) or the green fluorescent protein expressing plasmid (pEGFP-N1). Vessels were electroporated with eight 10 ms pulses of 200 V/cm. Sixty seconds after electroporation, the DNA solution was removed, the intestine returned to the abdomen and the abdominal wall closed with suture and metal wound clips. Six hours to 5 days later, rats were sacrificed and electroporated vessels were recovered. Luciferase activity of the blood vessels was monitored. Gene expression was detected as early as 6 h postelectroporation, peaked at 1-3 days with levels up to 1 ng of reporter gene product per vessel segment and returned towards baseline by day 5. Histological analysis of blood vessel segments revealed green fluorescent protein-positive cells throughout the thickness of the vessel wall (endothelial cells to adventitia). Responses of electroporated vessels to vasoconstricting stimuli were indistinguishable from those of control vessels at either 2 or 40 days posttreatment. The results of this study provide evidence that electroporation is an effective means for introducing naked DNA into the blood vessel wall and form the basis for future studies on targeted gene therapy to the intact vasculature. PMID- 11025401 TI - Blockade of smooth muscle cell migration and proliferation in baboon aortic explants by interleukin-1beta and tumor necrosis factor-alpha is nitric oxide dependent and nitric oxide-independent. AB - Interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha) are found in injured and atherosclerotic vessels and have been shown to influence smooth muscle cell (SMC) function in vitro. We have investigated the effects of IL-1beta and TNFalpha on SMC migration and proliferation in baboon aortic explants, an in vitro model of arterial injury. Because platelet-derived growth factor (PDGF) is also present in the vessel wall, we have studied the interaction of PDGF with the cytokines. IL-1beta and TNFalpha inhibited migration of SMCs and synthesis of DNA by SMCs. Cell death (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells and total DNA) was not altered by the cytokines. The cytokines increased levels of nitrite in the medium and L-nitroarginine partly reversed the inhibitory effects of the cytokines indicating a role for nitric oxide in these inhibitory effects. Treatment with indomethacin partially reversed the inhibition of migration, but not DNA synthesis by IL-1beta suggesting cyclooxygenase products play an inhibitory role in migration. PDGF-BB reversed the inhibitory effect of the cytokines on SMC migration, but not mitogenesis, without changing levels of nitrite in the medium. These data show that IL-1beta and TNFalpha decrease primate SMC migration and proliferation in arterial tissue partly through production of NO, and that PDGF antagonizes the effect of the cytokines. IL-1beta and TNFalpha may act directly to limit injury induced intimal hyperplasia by decreasing SMC migration and proliferation. PMID- 11025402 TI - Ascorbate transport in pig coronary artery smooth muscle: Na(+) removal and oxidative stress increase loss of accumulated cellular ascorbate. AB - Pig deendothelialized coronary artery rings and smooth muscle cells cultured from them accumulated ascorbate from medium containing Na(+). The accumulated material was determined to be ascorbate using high-performance liquid chromatography. We further characterized ascorbate uptake in the cultured cells. The data fitted best with a Hill coefficient of 1 for ascorbate (K(asc) = 22 +/- 2 microM) and 2 for Na(+) (K(Na) = 84 +/- 10 mM). The anion transport inhibitors sulfinpyrazone and 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS) inhibited the uptake. Transferring cultured cells loaded with (14)C-ascorbate into an ascorbate-free solution resulted in a biphasic loss of radioactivity - an initial sulfinpyrazone insensitive faster phase and a late sulfinpyrazone-sensitive slower phase. Transferring loaded cells into a Na(+)-free medium increased the loss in the initial phase in a sulfinpyrazone-sensitive manner, suggesting that the ascorbate transporter is bidirectional. Including peroxide or superoxide in the solution increased the loss of radioactivity. Thus, ascorbate accumulated in coronary artery smooth muscle cells by a Na(+)-dependent transporter was lost in an ascorbate-free solution, and the loss was increased by removing Na(+) from the medium or by oxidative stress. PMID- 11025403 TI - Myofibroblast involvement in glycosaminoglycan synthesis and lipid retention during coronary repair. AB - Myofibroblasts of adventitial origin have been linked to neointimal formation and remodeling after coronary injury. Accordingly, the goal of this study was to examine whether myofibroblasts contribute to focal accumulation of glycosaminoglycans (GAGs) and lipids during coronary repair. GAG synthesis was assessed by ex vivo labeling of balloon-injured porcine coronary arteries with (14)C-glucosamine. The synthesis of total GAGs transiently increased at 8 days in the normolipemic model (a 2.2-fold increase over baseline, p < 0.05). The majority of newly synthesized GAGs were sensitive to chondroitin ABC lyase (chondroitin/dermatan sulfate GAGs). Versican was localized to myofibroblast-rich regions in the adventitia and neointima [positive for alpha-smooth muscle (SM) actin, negative for h-caldesmon and SM myosin heavy chain]. In contrast, the adjacent SM-rich media showed no increase in versican expression. The association between injury-induced GAG accumulation and lipid retention was examined at 2 weeks after coronary injury in the hyperlipemic model. Lipid (Oil Red O) accumulated in the neointima and adventitia, but not in the adjacent media. Coronary repair under hyperlipemic conditions was associated with macrophage infiltration (19 +/- 5 vs. 3 +/- 2% of neointimal cells in normolipemic animals, p < 0.001) and increased neointimal formation (1.8 +/- 0.5 vs. 1.0 +/- 0.3 mm(2) in normolipemic animals, p < 0.01). In conclusion, this study demonstrated a transient increase in GAG synthesis following coronary injury. Chondroitin sulfate proteoglycans (e.g., versican) were rapidly synthesized by activated adventitial and neointimal cells which could contribute to early lipid retention in injured vessels. PMID- 11025404 TI - Microvascular endothelial cells differ in their basal and tumour necrosis factor alpha-regulated expression of adhesion molecules and cytokines. AB - We recently located a rare cytokeratin-positive (CK+) type of microvascular endothelial cell (MVEC) in the corpus luteum and aorta. Bovine corpus luteum MVEC are known to be involved in the cyclic accumulation of eosinophils and macrophages. Since leukocyte migration is specifically mediated by adhesion molecules and the release of cytokines, we compared the expression of these factors in basal and TNF-alpha-stimulated CK+ MVEC and in common cytokeratin negative (CK-) MVEC in order to obtain an initial insight into the functional capacities of CK+ MVEC. CK- MVEC revealed significantly higher basal RANTES mRNA expression than CK+ MVEC, and TNF- alpha up-regulated RANTES mRNA in both types of MVEC. Only resting and stimulated CK- MVEC expressed granulocyte-macrophage colony-stimulating factor mRNA. Both MVEC types expressed monocyte colony stimulating factor mRNA, but remained negative for eotaxin and interleukin (IL)-5 mRNA even after stimulation. Resting CK+ MVEC were positive for CD29, CD31, CD49a and CD49e, but expressed most of these antigens at a significantly lower density than did CK- MVEC. In contrast to CK- MVEC, CK+ MVEC failed to express CD49b or MHC class II. The activation of CK+ MVEC with TNF-alpha induced the expression of CD62P, but not of CD49b or MHC class II. In summary, phenotypically variable MVEC derived from the microvascular bed of one organ differ in their TNF-alpha regulated expression of cytokine mRNA and adhesion molecules. Morphological heterogeneity is related to a particular specialisation of functional MVEC. PMID- 11025405 TI - Characterization of the secreted form of endothelial-cell-specific molecule 1 by specific monoclonal antibodies. AB - Endothelial-cell-specific molecule 1 (ESM-1) is a recently identified endothelial cell molecule. As ESM-1 mRNA is preferentially expressed in human lung and kidney tissues, and as ESM-1 mRNA expression is regulated by inflammatory cytokines, ESM 1 is thought to play a role in the vascular contribution to organ-specific inflammation. In order to define its behavior, mouse anti-ESM-1 monoclonal antibodies were developed, and three distinct epitopes were mapped, which allowed development of a specific ELISA assay, immunohistological staining and immunoblot analysis. Here, we demonstrate that ESM-1 is present in cell lysates of human endothelial cells (human umbilical vein endothelial cells) with an apparent molecular weight of 20 kD. In contrast, the secreted form of ESM-1 is shifted to an apparent molecular weight of 50 kD, indicating that the secreted form of ESM-1 is posttranslationally modified. By ELISA, we show that the secretion of ESM-1 is significantly enhanced in the presence of TNFalpha. In contrast, the spontaneous as well as TNFalpha-induced secretion of ESM-1 is strongly inhibited by IFNgamma. Moreover, ESM-1 was detected in the serum of healthy subjects at an average concentration of 1.08 ng/ml, and we demonstrated that the serum level of ESM-1 is dramatically increased in patients presenting a septic shock. Analysis of ESM-1 expression in normal human tissues by immunohistochemistry showed that ESM-1 is localized in the vascular network, but also in the bronchial and renal epithelia. Our results demonstrate that ESM-1 is mainly expressed in the vascular endothelium both in vitro and in vivo, but also by different epithelia. ESM-1 may represent a new marker of endothelial cell activation, and may have a functional role in endothelium-dependent pathological disorders. PMID- 11025406 TI - Cytochrome P450 activity and endothelial dysfunction in insulin resistance. AB - Impaired endothelium-dependent relaxation attributable to nitric oxide/prostacyclin-independent factor (endothelium-dependent hyperpolarizing factor; EDHF) has been demonstrated in the small mesenteric arteries of insulin resistant rats. The purpose of this study was to determine if modulation of the cytochrome P450 enzyme system would restore EDHF-mediated relaxation in insulin resistant rats. Sprague-Dawley rats were randomized to control (n = 32) or insulin-resistant (n = 32) groups. Each group was further randomized to treatment (n = 48) or placebo (n = 16). Miconazole (3 days) and phenobarbital (3 and 14 days) achieved cytochrome P450 inhibition and induction, respectively. Following drug treatment, mean arterial pressure was measured and vascular function was assessed in small mesenteric arteries in vitro. Specifically, acetylcholine induced relaxation alone and in the presence of indomethacin plus N-nitro-L arginine (LNNA) or KCl was determined. Miconazole reduced the maximal relaxation in response to acetylcholine in control rats. Similarly, in the presence of LNNA plus indomethacin, acetylcholine-induced relaxation was impaired in the miconazole-treated control group versus the placebo group, whereas relaxation in the presence of KCl was unchanged. Miconazole did not affect relaxation in insulin-resistant arteries. In contrast, 3- and 14-day treatment with phenobarbital significantly improved acetylcholine-induced relaxation in insulin resistant arteries. Likewise, acetylcholine-mediated relaxation in the presence of LNNA plus indomethacin was also improved after phenobarbital treatment, while relaxation in the presence of KCl was unchanged. Phenobarbital treatment did not affect the control group. Miconazole treatment increased the mean arterial pressure in control rats, while 14-day phenobarbital treatment normalized the mean arterial pressure in insulin-resistant rats. Cytochrome P450 induction results in the restoration of EDHF-mediated relaxation in small mesenteric arteries and the normalization of mean arterial pressure in insulin-resistant rats. Thus, endothelial dysfunction secondary to insulin resistance can be reversed by the induction of cytochrome P450. PMID- 11025407 TI - Progesterone prevents estradiol-induced dendritic spine formation in cultured hippocampal neurons. AB - Estradiol has been shown to cause an increase in dendritic spine density in cultured hippocampal neurons, an effect mediated by downregulation of brain derived neurotrophic factor (BDNF) and glutamic acid decarboxylase (GAD), and the subsequent phosphorylation of cAMP response element binding protein (CREB) in response to enhanced activity levels. Interestingly, progesterone was shown to counteract the effects of estradiol on dendritic spine density in vivo and in vitro. The present study examined how progesterone may act to block the effects of estradiol in the molecular cascade of cellular events leading to formation of dendritic spines. Progesterone did not affect the estradiol-induced downregulation of BDNF or GAD, but it did block the effect of estradiol on CREB phosphorylation. The latter effects of progesterone on the pCREB response and spine formation were reversed by indomethacin, which prevents the conversion of progesterone to the neurosteroid tetrahydroprogesterone (THP). We therefore examined if the progesterone effects were caused by its active metabolite THP. Progesterone treatment caused a 60-fold increase in THP in the culture medium. THP itself enhanced spontaneous GABAergic activity in patch-clamped cultured neurons. Finally, THP blocked the estradiol-induced increase in spine density. These results suggest that progesterone, through conversion to THP, blocks the effects of estradiol on dendritic spines not via a direct nuclear receptor interaction but by counteracting the enhanced excitability produced by estradiol in the cultured network. PMID- 11025408 TI - Brain-derived neurotrophic factor but not neurotrophin-3 enhances differentiation of somatostatin neurons in hypothalamic cultures. AB - The present work investigated whether neurotrophins could differentially affect in vitro growth and maturation of two related subsets of hypothalamic neurons, hypophysiotropic somatostatin (SRIH) neurons projecting from the periventricular area and arcuate SRIH interneurons. For this purpose, the hypothalamus of 17-day old rat fetuses was sampled and separated into a ventral and a dorsal fragment containing respectively periventricular and arcuate regions. Each fragment was dissociated and seeded separately in defined medium. Brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3), two important members of the neurotrophin family involved in neuronal differentiation and plasticity, were added to the cultures at seeding time. After 6 or 11 days in vitro, neurons were labeled with an anti-SRIH antiserum and submitted to morphometric analysis. In parallel, SRIH mRNA was estimated by semiquantitative reverse-transcriptase-polymerase chain reaction, and neuronal SRIH content, basal and depolarisation-stimulated releases measured by radioimmunoassay. The response of control, non-labeled neurons was estimated by neuronal counts and by assaying glutamic acid decarboxylase, a marker of a large majority of hypothalamic neurons. BDNF markedly increased the size and the branching number of SRIH periventricular cell bodies. Expression of SRIH mRNA, as well as SRIH content and release into the culture medium, were also stimulated by the neurotrophin. Non-SRIH neurons were not affected by the treatment. Under the same conditions, arcuate neurons exhibited a weak, mostly transient response to BDNF. NT-3 was ineffective on either neuronal subset. Immunoneutralization of Trk receptors provided further evidence for BDNF effect specificity. The results indicate that BDNF is a selective activator of the differentiation of hypophysiotropic SRIH neurons in the periventricular area of the hypothalamus. PMID- 11025409 TI - Adrenergic and cholinergic regulation of in vitro melatonin release during ontogeny in the pineal gland of Long Evans rats. AB - Melatonin, produced by the pineal gland, plays an important role in a great variety of neuroendocrine functions. The rhythmic release of melatonin by the mammalian pineal gland is regulated by norepinephrine (NE) acting via alpha- and beta-adrenergic receptors utilizing distinct signal transduction pathways. Acetylcholine has been demonstrated to exert various effects in the mammalian pineal gland, including an inhibitory action on the NE-induced stimulation of melatonin production. However, data obtained by different laboratories on the interaction of adrenergic receptors are not consistent and whether muscarinic and/or nicotinic receptors participate in the various effects of acetylcholine is still contradictory. To investigate noradrenergic as well as cholinergic mechanisms during ontogeny, we have investigated in vitro melatonin release from isolated pineal glands of Long Evans rats of different ages. NE as well as the beta-adrenergic receptor agonist isoproterenol (ISO) significantly elevated the melatonin release in pineal glands from postnatal week 2 on. In pineal glands originating from 2- to 4-week-old rats, simultaneous activation of alpha- and beta-adrenergic receptors by ISO and the alpha-adrenergic receptor agonist methoxamine (MET) or NE resulted in significantly weaker stimulation of melatonin production than beta-receptor activation alone. Acetylcholine evoked a significant increase in melatonin release in pineal glands from 2- to 4-week-old rats. In pineal glands from 8- to 20-week-old animals, ISO, ISO + MET or NE stimulated pineal melatonin release to comparable maxima, whereas acetylcholine was without effect. Our data indicate (1) that the adrenergic stimulation of pineal melatonin production in Long Evans rats is dominated by a beta-adrenergic mechanism, (2) that additional alpha-adrenergic receptor activation is inhibitory and (3) dependent on the developmental status of the animal, and (4) that acetylcholine acting via muscarinic receptors has the capacity to stimulate melatonin release during early ontogeny. These data suggest that the melatonin generating system of the pineal gland of Long Evans rats undergoes substantial functional changes during early postnatal development, including adrenergic as well as cholinergic mechanisms. PMID- 11025410 TI - Effects of estradiol and raloxifene analog on brain, adrenal and serum allopregnanolone content in fertile and ovariectomized female rats. AB - Allopregnanolone is a neuroactive steroid synthesized in rat gonads, adrenal cortex, and central nervous system. It has been suggested that sex steroid hormones might influence allopregnanolone concentrations but no clear data have ever been reported. The aim of the present study was to investigate the effects of administration of 17beta-estradiol (17beta-E2), the raloxifene analog LY 117018 or their combination on allopregnanolone levels in fertile and ovariectomized (OVX) rats. Thirteen groups of 12 Wistar female rats each received either 17beta-E2 (0.1 or 1 microg/day) or LY-117018 (25, 250, and 1,250 microg/day), or 17beta-E2 1 microg/day plus LY-117018: 25, 250, and 1,250 microg/day for 14 days. The rats were then sacrificed and allopregnanolone content was assessed in the hypothalamus, hippocampus, pituitary, adrenals, and serum. Ovariectomy determined a significant decrease in allopregnanolone content in the hypothalamus, hippocampus, pituitary, and serum, while increasing it in the adrenals (p<0.01). In OVX rats, the administration of either 17beta-E2 or LY- 117018 restored ovariectomy-induced allopregnanolone changes. The administration of LY-117018 in addition to 17beta-E2 to OVX animals suppressed the increase in allopregnanolone levels determined by 17beta-E2 in the hippocampus, hypothalamus, and pituitary, but not in the adrenals and serum. In fertile rats, the administration of LY-117018 reproduced the effects of ovariectomy. This study shows that the raloxifene analog LY-117018 has an estrogen-like action on the central nervous system of OVX rats when administered alone, while it acts as an antiestrogen in the presence of 17beta-E2, both in OVX animals treated with 17beta-E2 and in fertile rats. A different effect was observed in the adrenal glands. The mechanism of action of this compound has still to be clarified. PMID- 11025411 TI - Interactions of IGF-1 with the blood-brain barrier in vivo and in situ. AB - Insulin-like growth factor-1 (IGF-1) given peripherally has been found effective in clinical trials to slow down neuronal degeneration in some nervous system diseases. This raises the question of whether and how IGF-1 crosses the blood brain barrier (BBB). In this report, we found that IGF-1 had a half-life of 4.5 min in blood, could remain intact for 20 min, and entered brain and spinal cord linearly. In the brain, IGF-1 had an influx rate of 0.4 microl/g x min after intravenous (iv) bolus injection as determined by multiple-time regression analysis. Intact radiolabeled IGF-1 was present in brain at 20 min after iv injection. Most of the injected IGF-1 entered the brain parenchyma instead of being entrapped in the cerebral vasculature. Addition of nonradiolabeled IGF-1 enhanced the influx of radiolabeled IGF-1 after iv injection, but inhibited the influx of radiolabeled IGF-1 by in-situ brain perfusion, suggesting that protein binding can explain the difference between the iv and perfusion experiments. In the spinal cord, the cervical region had the fastest uptake, followed by lumbar spinal cord. The thoracic spinal cord had the slowest uptake, comparable to that of brain. By contrast, des(1-3)IGF-1, an IGF-1 analogue with little protein binding but similar biological activity, had a shorter half-life in blood, slower influx rate into brain, and no alteration in pharmacokinetics after addition of nonradiolabeled peptide. We conclude that IGF-1 enters the CNS by a saturable transport system at the BBB, which functions in synchrony with IGF binding proteins in the periphery to regulate the availability of IGF-1 to the CNS. PMID- 11025412 TI - Effect of oxidative stress on the uptake of GABA and glutamate in synaptosomes isolated from diabetic rat brain. AB - It has been suggested that increased oxidative stress might be involved in the pathophysiology of diabetic complications. In this study, we investigated the effect of diabetes on the susceptibility of synaptosomes to oxidative stress (induced by the oxidizing pair ascorbate/Fe(2+)) and on the uptake of the amino acid neurotransmitters gamma-aminobutyric acid (GABA) and glutamate. We found a lower susceptibility of synaptosomes isolated from Goto-Kakizaki (GK) rats, a model of non-insulin-dependent diabetes mellitus, to lipid peroxidation as compared with synaptosomes isolated from Wistar control rats (6.40+/-1.05 and 12.14+/-1.46 nmol thiobarbituric acid reactive substance/mg protein, respectively). The lower susceptibility of GK rat synaptosomes to membrane lipid peroxidation correlates with an increase in synaptosomal vitamin E levels (835+/ 58.04 and 624.26+/-50.26 pmol/mg protein in diabetic and normal rats, respectively). In the absence of ascorbate/Fe(2+), no significant differences were observed between the levels of lipid peroxidation of synaptosomes isolated from diabetic and normal rats. Studies of neurotransmitter uptake show that the [(3)H]glutamate uptake was decreased by about 30% in diabetic GK rats as compared with control Wistar rats, whereas the [(3)H]GABA uptake was not significantly different from controls. Under oxidizing conditions, the glutamate uptake in diabetic rats was unaffected, and a decreased GABA uptake (41.39+/-4.41 and 60.96+/-6.4% of control in GK and Wistar rats, respectively) was observed. We conclude that the increased resistance to oxidative stress in GK rat synaptosomes may be due to the increased vitamin E content and that diabetic state and oxidative stress conditions differentially affected the uptake of the neurotransmitters GABA and glutamate. PMID- 11025414 TI - Cranial vault expansion: a comparison of magnetic coupled distraction to traditional surgical repositioning. AB - We compared the effectiveness of transcutaneously activated magnetic distraction of an osteotomized cranial bone flap to surgical repositioning of the flap with immediate, rigid internal fixation. Thirty immature rabbits were studied. All 30 rabbits underwent complete circumferential osteotomies of both parietal bones and postoperatively all were fitted with head frames. The rabbits were divided into 4 groups. The experimental magnetic distraction group consisted of 10 animals (group 1). In these rabbits magnets were secured to both parietal bones and magnets of opposite polarity were placed in the headframes. Each morning, the magnets in the head-frames were moved 0.25 mm farther away from the skull; this continued for 20 days for a total setback of 5 mm. Five animals served as the magnetic distraction controls; in these animals magnets were placed on the parietal bones but none were attached to the headframes (group 2). The experimental surgical repositioning group consisted of 10 rabbits (group 3). In each the parietal bones were elevated and fixed 5 mm above the rest of the skull using vitallium mesh and screws. Five rabbits made up the surgical respositioning control group (group 4). In these animals, parietal osteotomies were performed but the bones were secured to the skull in their original positions using hardware identical to that used in the experimental group. Six weeks later all of the animals were sacrificed. Two significant differences were identified between the experimental groups: (1) the cranial contours of the animals in the magnetic distraction group were rounded while those of the surgically repositioned group were acutely angled; (2) the osteotomies in rabbits in the magnetic distraction group were essentially completely ossified while in rabbits in the surgically repositioned group there were obvious gaps at the osteotomy sites that were filled with fibrous tissue. These conclusions support the potential utility of magnetically activated distraction as an alternative to current methods of cranial vault remodeling. PMID- 11025413 TI - Expression of opioid receptors in osteoblast-like MG-63 cells, and effects of different opioid agonists on alkaline phosphatase and osteocalcin secretion by these cells. AB - We have previously shown that several stressful situations associated with tissue injury determine a decrease in serum osteocalcin concentration. Since reduced osteocalcin production is a marker of decreased osteoblastic activity, this finding could be related to the pathogenesis of osteoporosis secondary to some diseases. Endogenous opioids are involved in stress response. Proenkephalin derived peptides have been shown to inhibit alkaline phosphatase activity, another marker of bone formation, in the murine cell line ROS-17/2.8. On the other hand, serum osteocalcin has been reported as being low in heroin abusers. We have therefore thought it of interest to study the presence of opioid receptors in the human osteoblast-like cell line MG-63, and to evaluate the effects of different opioid agonists on the secretion of alkaline phosphatase and osteocalcin by these cells. The presence of opioid receptors was studied by means of RT-PCR and immunohistochemistry. RT-PCR studies suggested the presence of specific mRNA for the three types of receptors, and immunohistochemistry clearly showed their occurrence. Osteocalcin synthesis was significantly inhibited by high concentrations of the mu agonists morphine and (D-Ala(2), N-MePhe(4),Gly(5) ol)-enkephalin although no changes were seen with the delta agonist (D-Ala(2),D leu(5))-enkephalin. Morphine-induced osteocalcin inhibition was abolished when osteoblastic cells were incubated simultaneously with naloxone, whereas it was potentiated when cells were preincubated with naloxone. None of the opioid agonists modified the secretion of alkaline phosphatase. In conclusion, human osteoblast-like cells MG-63 express the three types of opioid receptors. Endogenous opioids may be involved in the reduction of osteocalcin observed in stressful situations associated with tissue injury. PMID- 11025415 TI - Ventriculoperitoneal shunt flow dependency on the number of patent holes in a ventricular catheter. AB - Minimally invasive recanalization of obstructed ventricular catheters is a new treatment modality currently under investigation. It has been shown that blocked ventricular catheters can be at least partly reopened with ultrasonic cavitation, fiberoptic delivery of laser energy and electrocautery. It is not known, however, how many of the holes must be reopened in order to establish acceptable pressure flow characteristics. In this study, the pressure-flow relationship of partially obstructed ventricular catheters was determined using experimental and electrical circuit/computer models. It has been demonstrated in all cases that an additional pressure of no more than 0.5 cm of water is generated when only a single 500 microm-diameter hole in the ventricular catheter permits flow. Partial recanalization of a ventricular catheter provides acceptable CSF hydrodynamics. PMID- 11025416 TI - The prognosis of children with hydrocephalus and congenital heart disease. AB - The association of congenital hydrocephalus and heart disease in children is infrequent, but may present considerable dilemmas in management. This report describes the treatment and prognosis of 11 children with both clinical problems. There were 5 males and 6 females. Hydrocephalus occurred following aqueductal stenosis in 5 children and the Dandy-Walker malformation in 3. Three children were diagnosed with idiopathic hydrocephalus. Ten children underwent cerebrospinal fluid diversion procedures for control of hydrocephalus. Five children received pharmacological therapy for cardiac disease; 4 children required surgical correction. Two children died from medical conditions; 2 families declined treatment. Follow-up from 2 to 7 years in the remaining 7 children demonstrated moderate or severe neurodevelopmental disability in 5. One child at 2 years of age showed borderline developmental disability while 1 child is developing normally at 10 years of age. Overall the occurrence of symptomatic hydrocephalus and heart disease in the perinatal period resulted in mortality or neurodevelopmental disability in 9/11 children. PMID- 11025417 TI - Prolonged cerebral salt wasting syndrome associated with the intraventricular dissemination of brain tumors. Report of two cases and review of the literature. AB - Hyponatremia is a frequent event in neurosurgery practice and is usually associated with subarachnoid hemorrhage, head trauma, infections and neoplasms. The two common clinical manifestations are the inappropriate secretion of antidiuretic hormone (SIADH) and the cerebral salt wasting syndrome (CSWS), which were usually attributed to each other due to identical clinical presentation. In contrast to the better-recognized SIADH, there has not been a uniform consensus over the humoral and neural mechanisms of CSWS and functional aspects of renal response. In this article, we report on 2 cases of a primitive neuroectodermal tumor with prolonged CSWS manifested during the intraventricular dissemination of primary disease and the high catabolic stage. PMID- 11025418 TI - Giant intramedullary teratoma in an infant. AB - Teratomas are uncommon tumors of the nervous system, the commonest site being the pineal region. Spinal teratomas are very rare apart from the sacrococcygeal variety. The authors report the occurrence of an intramedullary teratoma in an infant. The clinical features, imaging studies and the pathology are discussed. In addition the literature is also reviewed. PMID- 11025419 TI - Endovascular treatment of huge cervicofacial hemangioma complicated by Kasabach Merritt syndrome. AB - A 2-month-old girl with a huge cervicofacial hemangioma complicated by Kasabach Merritt syndrome was presented. Two weeks corticosteroid treatment was ineffective, and deterioration of coagulation parameters and compromise of the airway and left vision due to huge hemangioma prompted use of endovascular treatment. Transfemoral arterial embolization using polyvinyl alcohol particles resulted in rapid clinical improvement. Endovascular treatment should be considered as an important therapeutic option for hemangioma complicated by Kasabach-Merritt syndrome. PMID- 11025420 TI - Interhemispheric neuroepithelial cyst associated with agenesis of the corpus callosum. A case report and review of the literature. AB - This report describes a neonatal case in whom a large interhemispheric cyst associated with agenesis of the corpus callosum was revealed by fetal ultrasonography and demonstrated by MRI to be multilobulated. Endoscopic fenestration of cysts was initially designed in view of the development of the patient's brain and surgical invasiveness. One year later, when motor paresis of the left arm and progressive enlargement of the cyst on MRI were revealed, open surgery was performed. The histological diagnosis was a neuroepithelial cyst with the feature of choroid plexus epithelia. The clinicopathological features of interhemispheric epithelial cysts associated with agenesis of the corpus callosum are reviewed in the light of differential diagnosis and therapeutic considerations. PMID- 11025421 TI - Critical shunt-induced subdural hematoma treated with combined pressure programmable valve implantation and endoscopic third ventriculostomy. AB - The authors present 2 patients with VP shunt-induced subdural hematomas (SDH) treated with pressure-programmable valve implantation and endoscopic third ventriculostomies (TV). The first patient is an 11-year-old girl who developed a shunt-induced SDH. Revision of the shunt valve with a higher-pressure valve resulted in a prolonged deterioration of her consciousness. External ventricular drainage at low pressure led to clinical improvement. A pressure-programmable valve set at 50 mm H(2)O was implanted, and the pressure gradually increased. At a pressure of 120 mm H(2)O symptoms recurred, even though the subdural collection was beginning to decrease in size. An endoscopic TV was performed, and the valve pressure was then increased to 200 mm H(2)O without any neurological symptoms. The second patient, a 7-year-old boy with shunt-induced SDH, had recurrent SDH, even after shunt revision with placement of a higher-pressure valve, which resulted in prolonged lethargy. A pressure-programmable valve was implanted with concurrent endoscopic TV. Gradual valve pressure increases up to 200 mm H(2)O could be performed without recurrent symptoms. Eventually, the shunt system was ligated to resolve residual positional headache, and the TV has been patent for more than 3 years. In both patients, the pressure-programmable valve was useful, since the optimal CSF drainage pressure changed during the period of recovery from symptomatic subdural collections. Concurrent TV appeared to enable increasing the valve pressure gradually without any neurological symptoms. The advantages of this combined approach are discussed. PMID- 11025422 TI - Cerebral aspergillosis in children. Report of three cases. AB - Between March and December of 1997, 3 cases of infantile cerebral aspergillosis appeared in our hospital. All patients presented some kind of immunosuppression associated with clinical findings of intracranial hypertension. After surgery, all cases were histopathologically confirmed and had positive cultures of Aspergillus. The first case was a patient with a history of subtotal resection of supratentorial ependymoma which presented a new enhancing mass on a control CT scan. Brain tumor recidiva was suspected, although, an Aspergillus granuloma was encountered on the tumor bed. Both other cases were found in patients who underwent transplantation: one presented multiple cerebral lesions after a renal transplant, and the other, after a bone marrow transplant, developed a rhinocerebral aspergillosis. The discussion was focused on the clinical findings, images and treatment of this cerebral infectious disease. PMID- 11025423 TI - Molecular events implicated in brain tumor angiogenesis and invasion. AB - We have conducted studies designed to help elucidate the molecular mechanisms involved in brain tumor invasion and angiogenesis, which are critical in the growth of malignant tumors of the central nervous system. A variety of molecular factors have been implicated in these processes. Here we focus on three that are of particular importance in the progression of brain tumors. Angiopoietins are involved in the regulation of vascular development. Hypoxia inducible factor-1 is a transcription factor that up-regulates genes, including genes encoding vascular endothelial growth factor under hypoxic conditions. Focal adhesion kinase is associated with infiltration of tumor cells and angiogenesis. PMID- 11025424 TI - Encephaloceles. PMID- 11025425 TI - Neoadjuvant hormonal therapy in radical prostatectomy and radiation-treated patients. AB - Neoadjuvant hormonal therapy (NHT) prior to radical prostatectomy or radiation therapy for clinically localized prostate cancer has regained interest in recent years but proof of its clinical benefit with respect to patient survival is still pending. We reviewed the literature regarding NHT in prostate cancer; only studies with a high evidence level were included for analysis. Five prospective randomized trials of NHT in radical prostatectomy-treated patients could be identified. Despite significantly lower numbers of positive surgical margins in the NHT group, prostate specific antigen failure rates were similar in the NHT group and the surgery only group. Data on disease-free and overall survival are not yet available. The two prospective randomized studies of NHT in radiation treated patients could demonstrate better local control of the tumor in the NHT group. However, there was no difference in overall survival between the NHT arm and the radiation only arm. Future clinical research should consider type and duration of hormonal treatment and evaluate potential surrogate end points. To date, there is no hard evidence in favor of NHT prior to radical prostatectomy or external beam irradiation. NHT in prostate cancer should only be applied within controlled trials. PMID- 11025427 TI - Antagonist/agonist balance of the nonsteroidal antiandrogen bicalutamide (Casodex) in a new prostate cancer model. AB - Androgen ablation is standard therapy for advanced prostate carcinoma. It can be administered either as a monotherapy or as a combined androgen blockade. In the present study we have investigated molecular mechanisms which are responsible for the development of resistance to therapy in advanced prostate cancer. For this purpose, we have cultured LNCaP cells in steroid-depleted medium for 1 year. The newly generated subline LNCaP-abl was characterized. In early passages (<75) LNCaP-abl cells showed a biphasic hypersensitive response to androgenic stimulation. Passages later than 75 are inhibited by androgen. Proliferation of LNCaP-abl cells was stimulated by the pure nonsteroidal antiandrogen bicalutamide (Casodex). To improve our understanding of changes which occur during intermittent androgen ablation, we have generated the sublines LNCaP-R (reversal; cultured with fetal calf serum) and LNCaP-RA (reversal and androgen; cultured with fetal calf serum and androgen) from LNCaP-abl cells. In both cell lines an increase of the basal proliferation rate was observed. Androgen receptor expression in LNCaP-abl cells was 4-fold higher than that in parental LNCaP cells (4.7 vs. 1.2 fmol/microg protein). Androgen receptor content in LNCaP-R cells was 1.8 fmol/microg protein and in LNCaP-RA cells 1.0 fmol/microg protein. The basal androgen receptor activity was 30-fold higher in LNCaP-abl cells compared to that in parental LNCaP cells. This basal activity was reduced in LNCaP-RA cells. Both androgen and the nonsteroidal androgen receptor antagonist hydroxyflutamide induced a 2- to 4-fold higher activation of androgen receptor in LNCaP-abl than in LNCaP cells. There was a switch from an antagonist to an agonist of the nonsteroidal antiandrogen bicalutamide (Casodex) in LNCaP-abl cells. Antagonistic properties of this androgen receptor blocker were again observed in both sublines (LNCaP-R and LNCaP-RA) derived from LNCaP-abl cells. In concordance with proliferation data in vitro, growth of LNCaP-abl cells in nude mice was stimulated by bicalutamide. In contrast, supplementation of androgen led to inhibition of proliferation of these cells. The present study provides new information that is useful for a better understanding of therapy-refractory prostate cancer. It is also important for the development of new therapy strategies for advanced carcinoma of the prostate. PMID- 11025426 TI - Expression of the ET(A) and ET(B) endothelin receptor subtype mRNA in human detrusor cultured smooth muscle cells. AB - The aim of this study was to determine the expression of the endothelin receptor subtype mRNAs in human detrusor cultured smooth muscle cells using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). First strand cDNA was made from human detrusor cultured smooth muscle cells total RNA and used for PCR with primers designed to amplify fragments of the ET(A) and ET(B) endothelin receptor subtype cDNA sequences. Subcloned fragments of the ET(A) and ET(B) endothelin receptor cDNAs were used to synthesize digoxigenin labeled cRNA probes by in vitro transcription. COS-7 cells transfected with the ET(A) and ET(B) receptor cDNAs were used as positive control and to confirm the absence of cross-hybridization due to sequence homology. Both ET(A) and ET(B) receptor mRNAs were detected by RT-PCR analysis. By ISH, both ET(A) and ET(B) receptor subtype mRNAs were detected. However, ET(A) signal was much more intense than ET(B) signal. These results indicate that mRNAs for both ET(A) and ET(B) receptors are expressed in detrusor smooth muscle cells of human urinary bladder. The ET(A) receptor is the predominant detrusor ET receptor. PMID- 11025428 TI - Is systematic sextant biopsy suitable for the detection of clinically significant prostate cancer? AB - BACKGROUND: The optimal extent of the prostate biopsy remains controversial. There is a need to avoid detection of insignificant cancer but not to miss significant and curable tumors. In alternative treatments of prostate cancer, repeated sextant biopsies are used to estimate the response. The aim of this study was to investigate the reliability of a repeated systematic sextant biopsy as the standard biopsy technique in patients with significant tumors which are being considered for curative treatment. METHODS: Systematic sextant biopsy was performed in vitro in 92 radical prostatectomy specimens. Of these patients, 81 (88.0%) had palpable lesions. RESULTS: Of the 92 investigated patients, 70 (76.1%) had potentially curable pT2-3pN0 prostate cancers. In these patients, the cancer was detected only in 72.9% of cases by a repeated in vitro biopsy. In the pT2 tumors, there was a detection rate of only 66.7%. CONCLUSIONS: This study underlines the fact that a considerable number of significant and potentially curable tumors remain undetected by the conventional sextant biopsy. A negative sextant biopsy does not rule out significant prostate cancer. PMID- 11025429 TI - Pathologic 'fused gland' as a prognostic factor for prostate cancer. AB - We examined the significance of a pathologic 'fused gland' according to the WHO Mostofi classification for predicting prognosis in hormonally treated prostate cancer. Of 284 newly diagnosed patients receiving hormone therapy, 60 patients had specimens with no 'medullary' or 'column-cord' elements. Twenty-five of these had a 'fused gland', while 35 did not. Survival of these patients was analyzed. Cause-specific survival of the patients with no medullary or column-cord elements showed a significantly better prognosis compared to others. Furthermore, the patients whose specimens contained a 'fused gland' component had significantly poorer survival than those without that component, suggesting that the 'fused gland' component is a poor prognostic factor for survival in patients with hormonally treated prostate cancer. PMID- 11025430 TI - Transurethral microwave thermotherapy for symptomatic benign prostatic hyperplasia: short-term experience with Prostcare. AB - PURPOSE: To assess our short-term experience with transurethral microwave thermotherapy (TUMT) for symptomatic benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: From August 1993 through July 1994, in total 65 patients with symptomatic BPH were enrolled into this study. The patients' ages ranged from 56 to 95 years with a mean of 70 years. Under local anesthesia with intraurethral instillation of Xylocaine jelly only, all patients received one session of TUMT for up to 60 min with Prostcare equipment. Uroflowmetry was performed and international prostatic symptom score (IPSS) determined before 3 and 6 months after TUMT for assessment of efficacy. All adverse events were recorded and evaluated for clinical relevance. RESULTS: At 3 and 6 months following TUMT, the mean IPSS decreased from 19.7 +/- 6.8 (baseline) to 12.8 +/- 8.2 (-46%) and to 15.5 +/- 9.0 (-21%), respectively; the maximal urine flow rate at 3 and 6 months increased from 9.1 +/- 4.8 ml/s (baseline) to 11.0 +/- 4.9 ml/s (+21%) and to 10.9 +/- 5.6 ml/s (+19%), respectively. During TUMT, burning sensation was the most frequent complaint (38.5%), followed by urethral discomfort (29.2%) and urgency (9.2%). Two patients (3.1%) interrupted TUMT, because of intolerable pain. Following TUMT micturition pain (73.8%) and gross hematuria (45.9%) were the most adverse events. Most of these adverse events disappeared within 2 weeks. One patient suffered from skin erosion over the penoscrotal junction 1 week later. None had retrograde ejaculation; 1 patient complained of erectile dysfunction. CONCLUSION: Although the efficacy of TUMT with Prostcare became less prominent 6 months after TUMT, TUMT was still a tolerable, safe alternative treatment of BPH, especially in patients who were not suitable for transurethral resection of the prostate or anesthesia. PMID- 11025431 TI - The 'Wing' versus the 'Vapor Cut' electrodes in transurethral electrovaporization resection of the prostate: comparative changes in safety parameters. AB - We compared the early safety features of two-band resection devices used in transurethral vaporization-resection of the prostate (TUVRP). Ninety patients with prostatic outflow obstruction were prospectively evaluated in two open studies for the treatment using the 'Wing' cutting electrode in group 1 (Richard Wolf, Germany) and the 'Vapor Cut' electrode in group 2 (Karl Storz, Germany). Patients in both groups were comparable in age, presentation, prostate size and blood parameters. Safety features evaluated included changes in serum hemoglobin, hematocrit, sodium concentration and incidence of side effects. The mean drop in hemoglobin, hematocrit and serum sodium 1 h postoperatively for the Wing group were 0.54 g/dl, 2.04 ml/dl and 0.084 mEq/l respectively compared to a mean drop of 1.1 g/dl, 3.07 ml/dl and 3.1 mEq/l in the Vapor Cut group. These differences were statistically significant (p = 0.004, 0.03 and <0.0001 respectively). The mean resection weight was 24.78 +/- 10.4 and 24.46 +/- 14.79 g in groups 1 and 2, respectively; however, the mean operation time was significantly longer in the Wing group (47.7 vs. 37.1 min, p = 0.003). We conclude that both loops were safe and efficacious in the treatment of patients with prostatic outflow obstruction. Safety features that are related to the vaporization effect are influenced by the speed of resection. PMID- 11025432 TI - BTA quantitative assay and NMP22 testing compared with urine cytology in the detection of transitional cell carcinoma of the bladder. AB - PURPOSE: Both BTA TRAK and NMP22 urine concentrations have shown a sensitivity superior to urine cytology in the detection of bladder cancer. We compared these tumor markers with urine cytology performed on 3 consecutive samples and evaluated by an expert cytopathologist. PATIENTS AND METHODS: The investigations were conducted on 94 patients undergoing a diagnostic cystoscopy for a high suspicion of bladder cancer (group 1) and on 102 patients with previous history of transitional cell carcinoma awaiting a follow-up cystoscopy (group 2). Biopsy specimens were obtained also from tumor negative patients. Immunoassays for BTA TRAK and NMP22 were carried out according to standard methods. The choice of the cut-off was based on the ground of sensitivity and specificity curves intersection. Urine cytology results were expressed as positive, negative and 'dubious'. RESULTS: Overall sensitivity was 56% for NMP22 (cut-off 11 U/ml) and 57% for BTA TRAK (cut-off 60 U/ml). When dubious results were considered as positive cases, urine cytology achieved a sensitivity of 73.3%. Assuming dubious cases as negative results, urine cytology sensitivity resulted 59.3%. When the 2 groups of patients were evaluated separately with different cut-off, there was no significant gain in sensitivity for BTA TRAK and NMP22 over urine cytology. CONCLUSIONS: Urine cytology performed on 3 samples showed the highest sensitivity and specificity. The diagnostic advantage of urine cytology over BTA TRAK and NMP22 was maintained when patients were stratified by tumor grade. PMID- 11025433 TI - Experience with ureteral stone management in 1,082 patients using semirigid ureteroscopes. AB - OBJECTIVES: To assess the efficacy and complications of ureteroscopic lithotripsy (URS) using semirigid ureteroscopes. METHODS: We retrospectively analyzed the records of 1,082 consecutive patients with ureteral stones who were treated with URS with or without auxiliary extracorporeal shock wave lithotripsy for stone fragments. The efficacy was estimated using the stone-free rate and efficiency quotient (EQ). RESULTS: The stone-free rates were 79.0, 90.4 and 93.2% for upper, middle and lower ureteral stones, respectively. The EQ was 0.49 for upper, 0.79 for middle and 0.87 for lower ureteral stones. Ureteral perforation occurred in 54 cases (5.0%), of which 13 cases (1.2%) required nephrostomy (n = 11, 1.0%) or open surgery (n = 2, 0.2%). CONCLUSIONS: URS is a reasonable procedure with minor complications for stones located in the lower and middle ureter, but cannot be recommended as a first-line treatment for upper ureteral stones. PMID- 11025434 TI - Comparison of symptoms, morphological, microbiological and urodynamic findings in patients with chronic prostatitis/pelvic pain syndrome. Is it possible to differentiate separate categories? AB - 164 patients with prostatitic symptoms were evaluated by segmented urinalysis and culture and classified according to the National Institutes of Health classification system: 64 patients (38%) suffered from chronic bacterial prostatitis; 12 (7%) from inflammatory chronic pelvic pain syndrome, and 92 (55%) from non-inflammatory chronic pelvic pain syndrome. Transrectal ultrasound of the prostate, uroflowmetry and measurement of residual urine were also performed. Complaints were recorded using a questionnaire. Our studies revealed that leukocytes in expressed prostatic secretions could be detected in only 24 and 36% of patients with positive bacterial or chlamydial culture. Complaints, ultrasound and urodynamic findings were similar in the 3 groups. Therefore the differential diagnosis and therapy, based on the results of the 4-glass test and cultures as well as on transrectal ultrasound of the prostate, seem to be difficult. PMID- 11025435 TI - Up, up and away with new markers and PSA. PMID- 11025436 TI - Sacral nerve stimulation for pain relief in interstitial cystitis. AB - A 60-year-old woman was treated for severe interstitial cystitis pain using sacral nerve stimulation. Pain and accompanying bladder dysfunction were improved by temporary and permanent sacral nerve stimulation. Six months after implantation of a sacral neuromodulator the patient is pain free and significantly improved on bladder dysfunction. Interstitial cystitis may be an indication for functional electrostimulation. PMID- 11025437 TI - Squamous cell carcinoma of the prostate without evidence of recurrence 5 years after operation. AB - A 54-year-old man underwent radical cystoprostatectomy after prostatic needle biopsy which revealed squamous cell carcinoma of the prostate. However, the positive surgical margin of the pubis suggested residual disease. He received chemotherapy with methotrexate, peplomycin and cisplatin (MPD regimen). He is at present still alive without evidence of recurrence 5 years after the operation. This suggests that MPD chemotherapy is effective for squamous cell carcinoma of the prostate. PMID- 11025438 TI - Regulation of muscle regulatory factors by DNA-binding, interacting proteins, and post-transcriptional modifications. AB - Skeletal muscle differentiation is influenced by multiple pathways, which regulate the activity of myogenic regulatory factors (MRFs)-the myogenic basic helix-loop-helix proteins and the MEF2-family members-in positive or negative ways. Here we will review and discuss the network of signals that regulate MRF function during myocyte proliferation, differentiation, and post-mitotic growth. Elucidating the mechanisms governing muscle-specific transcription will provide important insight in better understanding the embryonic development of muscle at the molecular level and will have important implications in setting out strategies aimed at muscle regeneration. Since the activity of MRFs are compromised in tumors of myogenic derivation-the rhabdomyosarcomas-the studies summarized in this review can provide a useful tool to uncover the molecular basis underlying the formation of these tumors. PMID- 11025439 TI - Regulation of human monocyte proMMP-9 production by fetuin, an endogenous TGF beta antagonist. AB - Members of the matrix metalloproteinase family of enzymes degrade specific components of the extracellular matrix. MMP-9 is a type IV/V collagenase necessary for normal osteogenesis and is increased in inflammatory and neoplastic conditions. In such destructive diseases as emphysema and arthritis, and in epithelial cancers, MMP-9 is produced by cells of the monocyte lineage. Fetuin, a prominent serum glycoprotein, binds to and inactivates TGF-beta family members and through this mechanism regulates osteogenesis (Binkert et al., 1999, J Biol Chem 274:28514-28520.). We studied the effects of TGF-beta1 and fetuin on proMMP 9 release by the human monocyte line THP-1. Exogenous TGF-beta1 stimulated proMMP 9 release by THP-1 cells, with half-maximal stimulation at approximately 0.01 ng/ml TGF-beta1. Human fetuin (0.5-2 microM) partially inhibited this stimulation. Human fetuin alone stimulated THP-1 monocyte proMMP-9 release, with half maximal stimulation at approximately 0.25 microM fetuin. Neutralizing antibody specific for TGF-beta1 also stimulated proMMP-9 release, suggesting that endogenously-derived TGF-beta1 has an inhibitory effect. In freshly isolated human peripheral blood monocytes, fetuin stimulated proMMP-9 release with a dose response curve similar to that observed in THP-1 cells. Human fetuin also activated proMMP-9 present in THP-1 conditioned medium. Taken together, these data suggest that under physiological conditions, fetuin facilitates matrix degradation by monocyte-derived MMP-9, both by opposing the autocrine inhibitory effect of endogenous TGF-beta1 on proMMP-9 release, and by activating proMMP-9 in the pericellular milieu. Conversely, fetuin may limit the stimulation of monocyte proMMP-9 release by high levels of exogenous TGF-beta1. Such modulation could prove important under pathological conditions where TGF-beta1 derived from paracrine sources is abundant, such as in epithelial malignancies. PMID- 11025441 TI - Expression and function of ryanodine receptors in human melanocytes. AB - We investigated the expression of ryanodine receptors (RyRs) in cultured human melanocytes with immunocytochemistry and reverse transcriptase-polymerase chain reaction. With the use of a monoclonal antibody, RyR immunoreactivity was detected in the cytoplasm of melanocytes, and was further confirmed by RT-PCR assay. The PCR products were cut with restriction enzymes specific for each RyR isoform. Using the RyR1-specific restriction enzyme SacI yielded fragments of 300, 100, and 130 base pairs, consistent with the expression of RyR1 isoforms. The function of RyR in Ca(2+) signaling was investigated using single-cell fura-2 imaging. Ryanodine (1 to approximately 100 microM) induced significant elevation of cytoplasmic Ca(2+) in single human melanocytes in a dose-dependent manner. The ryanodine-induced [Ca(2+)](i) increase was inhibited by neomycin. Furthermore, ryanodine inhibited proliferation and stimulated pigmentation of human melanocytes. This study demonstrates that the RyR1 isoform is expressed in cultured human melanocytes, and suggests that the RyR may be involved in regulating the intracellular Ca(2+) responses involved in proliferation and pigmentation of cultured human melanocytes. PMID- 11025440 TI - Role of the sarcoplasmic reticulum in regulating the activity-dependent expression of the glycogen phosphorylase gene in contractile skeletal muscle cells. AB - Nerve-evoked contractile activity in skeletal muscle regulates transcript and protein levels of many metabolic genes in a coordinate fashion, including the muscle isozyme of glycogen phosphorylase (MGP). Cellular signaling mechanisms mediating the activity-dependent modulation of MGP transcript levels were investigated in a spontaneously contractile rat skeletal muscle cell line (Rmo). Mechanisms regulating MGP mRNA levels in Rmo myotubes were compared with those previously shown to modulate the gene encoding the alpha subunit of the acetylcholine receptor (alphaAChR). Reducing the resting membrane potential from 78 to -30 mV, either electrochemically (KCl) or by increasing Na(+) permeability (veratridine): (1) prevented activation of transverse tubules, (2) impeded calcium release by the sarcoplasmic reticulum (SR), and (3) blocked Rmo contractility. MGP mRNA levels decreased to 30% of control levels and alphaAChR levels increased to 350% following 24 h of depolarization. Differing mechanisms appear to mediate this voltage-dependent regulation of MGP and alphaAChR. Inhibition of SR calcium efflux selectively decreased MGP mRNA levels by 30-50% when using dantrolene, thapsigargin, or a dose of ryanodine shown to inactivate Ca(2+)-induced SR Ca(2+) release (CICR). By contrast, blockade of voltage sensors in transverse tubules with nifedipine, a dihydroaminopyridine (DHAP) antagonist, selectively increased alphaAChR mRNA levels by twofold. These data indicate that the voltage-dependent regulation of AChR gene expression differs from that modulating the MGP gene. KCl-induced depolarization and dantrolene both inhibit pulsatile SR Ca(2+) efflux in Rmo myotubes, but by differing mechanisms. Depolarization and dantrolene comparably reduced MGP mRNA levels and decreased MGP transcript stability from a t(1/2) of 24 h to 14.5 and 16 h, respectively. Reduced transcript stability can account for the observed reduction in mRNA levels of MGP in noncontractile Rmo myotubes and could be a significant regulatory mechanism in skeletal muscle that coordinates the activity-dependent expression of MGP with other glycogenolytic genes. PMID- 11025442 TI - Regulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by bone resorptive factors in osteoblastic cells. AB - In addition to their stimulating function on osteoclastic bone resorption, bone resorptive factors may regulate proteinases and related factors in osteoblastic cells to degrade bone matrix proteins. This study investigated the regulation of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) by bone resorptive factors in the cultures of mouse osteoblastic MC3T3-E1 cells, mouse primary osteoblastic (POB) cells, and neonatal mouse calvariae. Expression of either MMP-2, -3, -9, -11, -13, and -14 or TIMP-1, -2, and -3 was detected in MC3T3-E1 cells and POB cells. When the bone resorptive factors parathyroid hormone, 1,25-dihydroxyvitamin D(3), prostaglandin E(2), interleukin 1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha) were added to the cell cultures, MMP-13 mRNA levels were found predominantly to increase by all resorptive factors in the three cultures. mRNA levels of either MMP-3 and -9 or TIMP-1 and -3 were found to increase mainly by the cytokines IL-1beta and TNF alpha. BB94, a nonselective MMP inhibitor, neutralized the (45)Ca release stimulated by these resorptive factors to an extent similar to that of calcitonin, strongly suggesting that bone resorptive factors function at least partly through MMP formation. We propose that MMP-13 mRNA expression in osteoblastic cells may play an important role in stimulating matrix degradation by both systemic and local resorptive factors, whereas either MMP-3 and -9 or TIMP-1 and -3 might modulate matrix degradation by local cytokines only. PMID- 11025444 TI - ACTH-regulated expression of vascular endothelial growth factor in the adult bovine adrenal cortex: a possible role in the maintenance of the microvasculature. AB - Endothelial cells lining vessels of endocrine tissues are fenestrated. Interactions with the local environment via either soluble factors or cell-cell interactions appear to govern this terminal endothelial differentiation. Adrenocorticotropin (ACTH) has previously been reported to modulate endothelial fenestration in the rat adrenal cortex. Since vascular endothelial growth factor (VEGF) has been characterized as a potent inducer of endothelial fenestration, we aimed to characterize the status of VEGF expression in the bovine adult adrenal cortex and asked whether ACTH may regulate VEGF expression. By immunohistochemical analysis, we observed VEGF expression in steroidogenic cells from both zona glomerulosa and zona fasciculata of the bovine adrenal cortex. Double-labeling experiments performed on isolated cells in primary culture revealed VEGF immunoreactivity, essentially colocalized with the Golgi apparatus. The expression of two predominant VEGF isoforms, VEGF(121) and VEGF(165), was observed by RT-PCR analysis. ACTH (10 nM) was found to rapidly (within 2-4 h) increase the abundance of these VEGF transcripts, as assessed by both RT-PCR and Northern blot analysis. In parallel, ACTH significantly induced VEGF secretion into the medium of fasciculata cells in primary culture. Thus, our data are consistent with the involvement of ACTH, through its regulation of VEGF expression, in the maintenance of the adult adrenal cortex endothelium. PMID- 11025443 TI - PKC and ERK1/2 regulate amylase promoter activity during differentiation of a salivary gland cell line. AB - The addition of transforming growth factor alpha (TGFalpha) to a human submandibular gland cell line (HSG) cultured on basement membrane extract Matrigel, synergistically activates the acinar cell-specific salivary amylase promoter. Signaling through beta1 integrins and increased phosphorylation of ERK1/2 are involved in the increased promoter activity. Phorbol-12-myristate-13 acetate (PMA) and thapsigargin increase amylase promoter activity, suggesting that phorbol ester and calcium-dependent protein kinase C (PKC) pathways are also involved. The combination of specific inhibitors of PKC and MEK1 inhibits the amylase promoter. Inhibitors of the calcium-dependent PKC isoforms alpha, beta, and gamma decrease the promoter activity; however, PKCbeta is not detectable in HSG cells. TGFalpha alters the cellular localization of PKCalpha but not -gamma, suggesting PKCalpha is involved in TGFalpha upregulation of the amylase promoter. Furthermore, rottlerin, a PKCdelta-specific inhibitor, increases the promoter activity, suggesting PKC isoforms differentially regulate the amylase promoter. In conclusion, beta1-integrin and TGFalpha signaling pathways regulate the amylase promoter activity in HSG cells. In response to Matrigel and TGFalpha, the activation of both PKCalpha and phosphorylation of ERK1/2 results in synergistic activation of the amylase promoter. Published 2000 Wiley-Liss, Inc. PMID- 11025445 TI - Independent activation of endogenous p21-activated protein kinase-3 (PAK3) and JNK by thrombin in CCL39 fibroblasts. AB - Thrombin, a potent mitogen for CCL39 hamster lung fibroblasts, activates the seven membrane-spanning receptor PAR1. To better understand the signaling pathways controlled by this receptor we analyzed a potential downstream effector, p21-activated protein kinase (PAK). Thrombin and PAR1 agonist peptide, as well as serum and lysophosphatidic acid, were found to stimulate HA-mPAK3 activity in CCL39 cells transfected with a plasmid encoding the epitope-tagged kinase. Similar results were obtained using antibodies developed against the endogenous kinase. PAK3 activation is sensitive to pertussis toxin, but insensitive to LY 294002, an inhibitor of phosphatidylinositol 3'-kinase. Thrombin and serum also activate c-jun amino terminal kinase (JNK). Similar to PAK3 activation, thrombin stimulated JNK activity is inhibited by pertussis toxin, but not by LY 294002. In a CCL39-derived cell line expressing constitutively active mPAK3 in a tetracyline dependent manner, induction of PAK activity does not lead to corresponding increases in JNK activity. Our findings indicate that PAK3 is responsive to thrombin and other G protein-coupled receptor systems. Furthermore, our data suggest that in CCL39 cells, JNK activation by thrombin occurs independently of PAK3. PMID- 11025446 TI - Differential regulation of three glucose transporter genes in a renal epithelial cell line. AB - Three hexose transporter genes, the Na(+)/glucose cotransporters SGLT1 and SGLT3 (formerly SAAT1/pSGLT2) and the facilitative transporter GLUT1, are expressed in a renal epithelial cell line with proximal tubule characteristics. A number of studies have demonstrated that SGLT1 expression is coupled to the cellular differentiation state and is also negatively regulated by its substrate glucose. In the present study, we demonstrate that SGLT3 mRNA expression is relatively unaffected by conditions promoting dedifferentiation (reseeding to a subconfluent density, activation of protein kinase C) or differentiation (confluent cell density, activation of protein kinase A) nor was expression sensitive to hyperglycemic glucose levels in the medium. We further demonstrate that protein kinase A and protein kinase C exert opposing effects on GLUT1 and SGLT1 mRNA levels in polarized cell monolayers, indicating that GLUT1 mRNA is also highly regulated in polarized epithelial cells by agents affecting cell differentiation. The relatively constitutive expression of SGLT3 mRNA suggests a novel role for this low-affinity Na(+)/glucose cotransporter, to provide concentrative glucose uptake under hyperglycemic conditions where expression of high-affinity glucose cotransporter SGLT1 mRNA is significantly downregulated. PMID- 11025448 TI - Translocation of FGF2 to the cell surface without release into conditioned media. AB - Like most cells in culture, stably transfected COS-1 cells (CF18) that constitutively overexpress basic fibroblast growth factor (FGF2) do not release the growth factor into conditioned media. Yet, when cells were biotinylated, 30% of the total cell-associated immunoreactive FGF2 was detected on the cell surface. Under similar conditions, up to 70% of the total immunoreactive FGF2 in transfected endothelial cells (MAE ZIP) or untransfected rat (C6) and human (U87MG) glioblastoma cell lines was detected on their cell surface. When peripheral plasma membrane proteins were removed from the cell surface with 0.1 M sodium carbonate, the amount of exported FGF2 was significantly reduced, whereas cell viability was unaffected. FGF2 then reappeared on the cell surface in a time dependent manner. Ouabain, a cardenolide previously shown to inhibit the export of FGF2 from transiently transfected COS-1 cells, blocked the appearance of FGF2 onto the surface of transfected CF18 cells and MAE ZIP cells but had no detectable effect on C6 and U87MG cells. The observation that the translocation of FGF2 onto the cell surface is dissociated from its release into conditioned medium is consistent with FGF2's being rarely found in biological fluids but always cell associated and in the extracellular matrix. The findings point to a role played by the protein export pathway in controlling FGF2 activity and the normal physiological function that this growth factor plays in cell growth and differentiation. The widely accepted presumption that the absence of FGF2 in conditioned media reflects its inability to exit the cell needs to be reevaluated. PMID- 11025447 TI - Apoptosis of lung epithelial cells in response to TNF-alpha requires angiotensin II generation de novo. AB - Recent work from this laboratory demonstrated that apoptosis of pulmonary alveolar epithelial cells (AEC) in response to Fas requires angiotensin II (ANGII) generation de novo and binding to its receptor (Wang et al., 1999b, Am J Physiol Lung Cell Mol Physiol 277:L1245-L1250). These findings led us to hypothesize that a similar mechanism might be involved in the induction of AEC apoptosis by TNF-alpha. Apoptosis was detected by assessment of nuclear and chromatin morphology, increased activity of caspase 3, binding of annexin V, and by net cell loss inhibitable by the caspase inhibitor ZVAD-fmk. Purified human TNF-alpha induced dose-dependent apoptosis in primary type II pneumocytes isolated from rats or in the AEC-derived human lung carcinoma cell line A549. Apoptosis in response to TNF-alpha was inhibited in a dose-dependent manner by the nonselective ANGII receptor antagonist saralasin or by the nonthiol ACE inhibitor lisinopril; the inhibition of TNF-induced apoptosis was maximal at 50 microgram/ml saralasin (101% inhibition) and at 0.5 microgram/ml lisinopril (86% inhibition). In both cell culture models, purified TNF-alpha caused a significant increase in the mRNA for angiotensinogen (ANGEN), which was not expressed in unactivated cells. Transfection of primary cultures of rat AEC with antisense oligonucleotides against ANGEN mRNA inhibited the subsequent induction of TNF stimulated apoptosis by 72% (P < 0.01). Exposure to TNF-alpha increased the concentration of ANGII in the serum-free extracellular medium by fivefold in A549 cell cultures and by 40-fold in primary AEC preparations; further, exposure to TNF-alpha for 40 h caused a net cell loss of 70%, which was completely abrogated by either the caspase inhibitor ZVAD-fmk, lisinopril, or saralasin. Apoptosis in response to TNF-alpha was also completely inhibited by neutralizing antibodies specific for ANGII (P < 0.01), but isotype-matched nonimmune immunoglobulins had no significant effect. These data indicate that the induction of AEC apoptosis by TNF-alpha requires a functional renin/angiotensin system (RAS) in the target cell. They also suggest that therapeutic control of AEC apoptosis in response to TNF-alpha is feasible through pharmacologic manipulation of the local RAS. PMID- 11025449 TI - Molecular characterization of celtix-1, a bromodomain protein interacting with the transcription factor interferon regulatory factor 2. AB - Transcriptional control at the G1/S-phase transition of the cell cycle requires functional interactions of multimeric promoter regulatory complexes that contain DNA binding proteins, transcriptional cofactors, and/or chromatin modifying enzymes. Transcriptional regulation of the human histone H4/n gene (FO108) is mediated by Interferon Regulatory Factor-2 (IRF-2), as well as other histone gene promoter factors. To identify proteins that interact with cell-cycle regulatory factors, we performed yeast two-hybrid analysis with IRF-2 and identified a novel human protein termed Celtix-1 which binds to IRF-2. Celtix-1 contains several phylogenetically conserved domains, including a bromodomain, which is found in a number of transcriptional cofactors. Using a panel of IRF-2 deletion mutants in yeast two-hybrid assays, we established that Celtix-1 contacts the C-terminus of IRF-2. Celtix-1 directly interacts with IRF-2 based on binding studies with glutathione S-transferase (GST)/IRF-2 fusion proteins, and immunofluorescence studies suggest that Celtix-1 and IRF-2 associate in situ. Celtix-1 is distributed throughout the nucleus in a heterodisperse pattern. A subset of Celtix-1 colocalizes with the hyperacetylated forms of histones H3 and H4, as well as with the hyperphosphorylated, transcriptionally active form of RNA polymerase II. We conclude that the bromodomain protein Celtix-1 is a novel IRF-2 interacting protein that associates with transcriptionally active chromatin in situ. PMID- 11025450 TI - A new apoptotic pathway for the complement factor B-derived fragment Bb. AB - Apoptosis is involved in both the cellular and humoral immune system destroying tumors. An apoptosis-inducing factor from HL-60 myeloid leukemia cells was obtained, purified, and sequenced. The protein found has been identified as a human complement factor B-derived fragment Bb, although it is known that factor B is able to induce apoptosis in several leukemia cell lines. Monoclonal antibodies against fragment Ba and Bb inhibited the apoptotic activity of factor B. When the purified fragment Bb was used for apoptosis induction, only the anti-Bb antibody inhibited Bb-induced apoptosis, and not the anti-Ba antibody. The apoptosis inducing activity was found to be enhanced under conditions facilitating the formation of Bb. Blocking TNF/TNFR or FasL/Fas interactions did not interfere with the factor B-induced apoptosis. CD11c (iC3bR) acts as the main subunit of a heterodimer binding to fragment Bb in the apoptosis pathway, and the factor B derived fragment Bb was found to possess the previously unknown function of inducing apoptosis in leukemic cells through a suicide mechanism of myeloid lineage cells during the differentiation stage. PMID- 11025451 TI - Induction of MRP1 and gamma-glutamylcysteine synthetase gene expression by interleukin 1beta is mediated by nitric oxide-related signalings in human colorectal cancer cells. AB - Treatment of human colorectal cancer cells HT29 with interleukin 1beta (IL-1beta) induces expression of the multidrug resistance protein (MRP1) gene encoding the ATP-dependent glutathione S-conjugate export (GS-X) pump and the gamma glutamylcysteine synthetase (gamma-GCSh) gene encoding heavy (catalytic) subunit of gamma-glutamylcysteine synthetase, the rate-limiting enzyme for the biosynthesis of glutathione (GSH). The induction can be suppressed by N(G)-methyl L-arginine, a specific inhibitor of nitric oxide synthase (NOS). These results suggest that IL-1beta-mediated MRP1 and gamma-GCSh induction involve nitric oxide (NO) -related signaling. Further supports to the involvement of NO in the induction of MRP1 and gamma-GCSh expression are made by the following observations. (i) Expression of MRP1 and gamma-GCSh genes were induced by treating the cells with NO donors, i.e., S-nitro-N-acetyl-D,L-penicillamide (SNAP) and S-nitroso-L-glutathione, in a concentration-dependent manner. (ii) Ectopic expression of inducible NOS (iNOS) activity by transfecting expressible recombinant iNOS cDNA encoding functional iNOS but not the nonfunctional version resulted in elevated expression of MRP1 and gamma-GCSh. We also demonstrated that HT-29 cells treated with either 1L-1beta or SNAP induced ceramide production, and addition of C2 or C6 ceramides into cultured HT-29 cells resulted in induction of gamma-GCSh but not MRP1 expression. Collectively, our results demonstrate that induction of MRP1 and gamma-GCSh by IL-1beta is regulated, at least in part, by an NO-related signaling, and induction of gamma-GCSh is by NO-related ceramide signaling. PMID- 11025452 TI - Retinol conversion to retinoic acid is impaired in breast cancer cell lines relative to normal cells. AB - The bioactivity of retinol (vitamin A) is in part dependent on its metabolism to retinoic acid (RA). We investigated the ability of breast epithelial cells to synthesize RA when challenged with a physiological retinol dose (2 microM). Normal human mammary epithelial cells (HMEC) cultured from reduction mammoplasties were competent in RA synthesis and the ability to synthesize RA was retained by immortal, nontumorigenic breast epithelial cell lines (MTSV1.7, MCF 10F, and 184B5). In contrast, most (five of six) breast cancer cell lines could not synthesize RA or did so at low rates relative to normal cells. A notable exception was the MDA-MB-468 cell line, which was fully competent in RA synthesis. Most (>/=68%) of the RA synthesized by breast cells was recovered from the culture medium. Cellular retinol binding protein and cellular RA binding protein II, both expressed in HMEC, had various expression patterns in the cell lines that did not correlate with the observed differences in RA synthesizing ability. Strong RA induction of the RA hydroxylase P450RAI (CYP26) was confined to ERalpha-positive T47D and MCF-7 breast cancer cells and did not appear to explain the lack of detectable RA levels in these cells since RA remained undetectable when the cells were treated with 5-10 microM liarozole, a P450RAI inhibitor. We hypothesize that retinol bioactivity is impaired in breast cancer cells that cannot synthesize RA. In preliminary support of this hypothesis, we found that retinol (0.5-2 microM) inhibited MCF-10F but not T47D or MCF-7 cell growth. PMID- 11025453 TI - Regulation of sialomucin complex/Muc4 expression in rat uterine luminal epithelial cells by transforming growth factor-beta: implications for blastocyst implantation. AB - Blastocyst implantation is arguably the most critical stage of mammalian embryogenesis and requires that the uterus be in a receptive state. Initiation of receptivity involves loss of anti-adhesive molecules from the apical surface of the uterine luminal epithelium, one of which is sialomucin complex (SMC/Muc4), a highly O-glycosylated, anti-adhesive glycoprotein composed of mucin ascites sialoglycoprotein-1 (ASGP-1) and transmembrane (ASGP-2) subunits. SMC expression at the uterine luminal surface, but not in glandular epithelium, is hormonally regulated and varies with the estrous cycle. SMC is lost from the luminal uterine surface at the period of receptivity. However, the mechanism by which SMC is hormonally regulated is not understood. Analyses of SMC regulation in hormone responsive primary cultures of rat uterine luminal epithelial cells (RULEC) demonstrated robust SMC expression by the RULEC, which is not altered by treatments with estrogen or progesterone. However, both SMC protein and transcript are downregulated by transforming growth factor-beta (TGF-beta1). SMC is also downregulated when RULEC are co-cultured with isolated uterine stromal cells. Estradiol and anti-TGF-beta block the stromal cell effect. These results suggest an indirect hormonal regulation of RULEC SMC, in which TGF-beta acts as a hormonally regulated, mesenchymal paracrine factor to repress SMC production by the epithelial cells and permit implantation. PMID- 11025454 TI - Breast cancer: which paradigm? PMID- 11025455 TI - Internal hemipelvectomy for solitary pelvic thyroid cancer metastases. AB - BACKGROUND AND OBJECTIVES: Radioactive iodine (RAI) therapy remains a primary treatment modality for metastatic thyroid carcinoma, but poor tumor uptake of the agent can limit its usefulness. While offering effective palliation, radiation therapy is not curative, and chemotherapy is even less useful. Surgical resection occasionally remains the only hope of offering a long-term cure in the case of isolated metastases. METHODS: We describe 3 cases of thyroid cancer metastatic to the pelvic girdle that were unresponsive to RAI and other nonoperative therapies, which presented over a 15-year time period. The pelvic disease was the only site of active disease in all cases, and all 3 patients were suffering considerable pain. All 3 patients underwent internal hemipelvectomy with reconstruction in 2 cases using a pelvic bone allograft. RESULTS: All 3 experienced symptomatic relief and early mobilization. While the infection rate was 100%, these all responded completely to operative debridement, irrigation, and antibiotics. One patient, found postoperatively to have a positive surgical margin, developed a local recurrence at 4 months and died. A second patient developed a local recurrence at 11 months and died. The third patient underwent a revision of her hip arthroplasty because of acetabular loosening after a fall 21 months postoperatively. She is alive, disease-free, and ambulatory with the aid of a cane 32 months after the original procedure. CONCLUSIONS: We propose this surgical procedure in selected patients with metastatic pelvic thyroid cancer. It provides symptomatic relief with a chance for prolonged disease-free survival some patients. PMID- 11025456 TI - Malignancy in HIV/AIDs: a single hospital experience. AB - BACKGROUND AND OBJECTIVES: Our hospital serves an area with a significant number of patients seropositive for the human immunodeficiency virus (HIV). Intravenous drug abuse and heterosexual exposure are by far the predominant risk factors for HIV and acquired immunodeficiency syndrome (AIDS). Seven percent of these patients develop malignancies. Our aim was to study the types of tumor, their distribution, and to evaluate the patients' outcome. METHODS: Of 3,578 patients with HIV infection or AIDS treated between 1993 and 1998, 245 had 1 or more malignancies. Information was collected on age, sex, race, predisposing risk factors for AIDS, malignancies, symptoms at presentation, the time of the onset of AIDS, CD4 cell counts, pathology findings, and mortality. RESULTS: Although aspects of our patients resembled those of previously studied groups of patients with AIDS, there also were ways in which our patients differed from those other groups. Of our patients, 21. 6% had non-AIDS-defining (NAD) invasive malignancies. This was considerably higher than the rate in most studies. Twenty seven patients with such malignancies died during the study. Forty-two other patients had pre-invasive cancers. Among patients having AIDS-defining (AD) malignancies, 55.9% died, a fact that was related to patients' low CD4 cell counts and late presentation. Our 97 patients with Kaposi sarcoma included 22 women, a relatively high number that may be related to the fact that most of our patients were intravenous drug abusers or had become infected by heterosexual transmission of HIV. CONCLUSIONS: AIDS is associated with a high risk of malignancy and an unusual spectrum of tumors. Patients with invasive tumors have advanced disease at the time of initial presentation. Those with AD tumors have a worse prognosis than patients with NAD tumors. The impact of highly active antiretroviral therapy on both AD and NAD tumors needs to be further evaluated. PMID- 11025457 TI - Surgical management of epithelial ovarian cancer at community hospitals: A population-based study. AB - BACKGROUND AND OBJECTIVES: Accurate surgical staging and maximal tumor reduction are the basic management principles of epithelial ovarian cancer (EOC). The purpose of our study is to report on staging practices and the primary surgery of EOC in a region that has no tertiary oncological referral center and no surgical gynecological oncologist. METHODS: Between 1 January 1989 and 30 December 1995, the Valais Cancer Registry had registered 157 patients with ovarian cancer stage I-IV. Hospital case notes were reviewed retrospectively and patients who did not have a surgical abdominal exploration (n = 20), with borderline (n = 12) or non epithelial tumors (n = 13), operated upon in other regions (n = 8) and without complete medical records (n = 2) were excluded. Therefore 102 patients were evaluated. RESULTS: The interventions have been performed in 7 regional hospitals and 1 private clinic by 24 obstetricians-gynecologists and 8 general surgeons. In early EOC, 9% random peritoneal biopsies and 3% retroperitoneal lymph node samplings have been performed. In advanced EOC, 40% of patients had total abdominal hysterectomy, bilateral salpingo-oophorectomy and omentectomy and 42% had cytoreductive surgery with a residual tumor of 12 months was better than that of those with the interval 1.0 cm after primary operation should be considered prudently. PMID- 11025459 TI - Postsurgical sequential methotrexate, fluorouracil, and leucovorin for stages 3 and 4 gastric carcinoma: A preliminary study. AB - BACKGROUND AND OBJECTIVES: The present study compared the effects of sequential methotrexate and fluorouracil followed by leucovorin rescue (MFL), as an adjuvant chemotherapy vs. UFT (a combination of uracil and tegafur), on patient survival and recurrence following surgery for advanced gastric carcinoma. METHODS: Between July 1990 and June 1998, a total of 54 patients with advanced gastric cancer were treated postoperatively by adjuvant chemotherapy using MFL or UFT. Surgical treatment was performed according to standardized procedures for radical resection of gastric cancer. The patients were stratified into two groups following surgery. The MFL regimen consisted of methotrexate (100 mg/m2) and 5 fluorouracil (600 mg/m2) at hour 3, followed by leucovorin rescue. The oral UFT (375 mg/m2/day), a combination of tegafur and uracil in a molar ratio of 1:4, was continued for 3 years or longer depending on the patients tolerance. RESULTS: In stage 3 gastric cancer, the overall survival rates following surgery was significantly (p < 0.05) higher in the MFL than the UFT group. Difference in disease free survival was not statistically significant between the groups. Recurrence rates showed a trend (p = 0.08) to decrease in the MFL than the UFT group. In stage 4 gastric cancer, no significant difference was obtained in the overall survival rates between the groups. CONCLUSIONS: The present results suggested the superiority of MFL treatment for improving postoperative survival in patients with advanced gastric cancer, in particular for those patients with a high risk of recurrence following potential curative resection. In patients with stage 4 gastric cancer, however, MFL treatment showed similar effects as UFT on the postsurgical survival of the patients. PMID- 11025460 TI - Cervical lymph node metastasis in patients with submucosal carcinoma of the thoracic esophagus. AB - BACKGROUND AND OBJECTIVES: The effect of esophagectomy with three-field lymph node dissection for submucosal carcinoma of the thoracic esophagus remains controversial. The aim of this study was to evaluate the effect of esophagectomy with three-field lymph node dissection for submucosal carcinoma of the thoracic esophagus. METHODS: From January 1983 to December 1997, the records of 101 consecutive patients who underwent transthoracic esophagectomy with three-field lymph node dissection were retrospectively analyzed. RESULTS: The incidence of the operative complications was 70%. The 30-day and overall hospital mortality rates were 1.0% and 2.0%, respectively. The positive rate of histological cervical nodal metastasis was 17%. The 5-year survival rates for the patients with and those without cervical nodal metastasis were 55% and 71%, respectively. The difference between patients with and those without cervical nodal metastasis was not statistically significant. Cumulative 5-year survival rates for the patients with metastasis in the cervical, upper mediastinal, or abdominal lymph nodes were 55%, 65%, and 46%, respectively. There was no statistically significant difference between each survival. CONCLUSIONS: Three-field lymph node dissection may be indicated for patients requiring esophagectomy for submucosal carcinoma of the thoracic esophagus because the frequency of cervical lymph node metastasis is not negligible and acceptable overall hospital mortality and favorable survival rates of patients with histologically positive cervical nodes can be achieved. PMID- 11025461 TI - Clinicopathologic features and treatment of postirradiation sarcoma of bone and soft tissue. AB - BACKGROUND AND OBJECTIVES: An analysis of the clinicopathologic features and treatment of patients with postirradiation sarcoma of bone and soft tissue was performed to guide modern evaluation and management. METHODS: A retrospective analysis of 135 sarcomas in 130 patients was performed. RESULTS: The mean age of the 130 patients was 48 years, and there was a female predominance because of irradiation for carcinomas of the breast and uterus. Indication for irradiation was a soft tissue lesion (such as lymphoma or breast cancer) in 58.5% of patients and a bone lesion (such as giant cell tumor or fibrous dysplasia) in 41.5%. The latent period (interval between irradiation and discovery of the sarcoma) ranged from 4-55 years (mean, 17 years). Of the lesions, 74% were stage IIB (high-grade extra-compartmental) and 24% were stage III (metastases). Ninety-four patients received their treatment at our institution. Of the 61 patients with resectable disease, 49 had amputations and 12 had limb salvage procedures. The 5-year cumulative survival rate was 68.2% for patients with peripheral (extremities, including proximal femur and hip) resectable lesions and 27.3% for patients with central (pelvis, head/neck, and ribs) resectable lesions. The local recurrence rate correlated with the surgical margin achieved: intralesional, 73%, marginal, 64%, and wide, 23%. CONCLUSIONS: The prognosis for patients with peripheral resectable postirradiation sarcomas is good if a wide surgical margin can be achieved. This group of patients should be treated aggressively because they have a new cancer. PMID- 11025463 TI - Primary diffuse malignant peritoneal mesothelioma: case report and update of therapy. AB - BACKGROUND AND OBJECTIVES: Primary Diffuse Malignant Peritoneal Mesothelioma is a rare disease, with an incidence of 2.2 cases in 1. 000.000 in the USA. It occupies 10% of all mesotheliomas referred in literature. METHODS: We describe a case of diffuse malignant peritoneal mesothelioma arising in a 54-year-old woman who presented a small bowel occlusion. A middle line laparotomy was done; multiple biopsies and an ileostomy were performed. There was not a history of exposure to asbestos. Histologic diagnosis was based on light microscopy, histochemistry, and immunohistochemistry. RESULTS: Patient had no further treatment because of her poor general conditions. She died 4 months later. CONCLUSIONS: Update of treatment is briefly described with particular attention to multimodality approach (surgery, chemotherapy, radiotherapy) and other new therapeutic options (iperthermochemotherapy, immunotherapy, gene therapy), currently in clinical trials. PMID- 11025462 TI - The influence of enhanced postoperative inflammation by the intrapleural administration of streptococcal preparation (OK-432) on the prognosis of completely resected non-small-cell lung cancer. AB - BACKGROUND AND OBJECTIVES: It is not clear whether postoperative inflammation affects the prognosis of malignant disease. METHODS: We retrospectively reviewed the patients with non-small-cell lung cancer who underwent a complete resection at the National Kyushu Cancer Center from 1989 to 1996. For the treatment of prolonged air leakage after a pulmonary lobectomy, 25 patients received an intrapleural injection of OK-432, a lyophilized preparation of the heat- and penicillin-treated Su-strain of the Streptococcus pyogenes group A3. All patients were males who were older than 50 years of age. As a control, we selected 164 male patients who were older than 50 years of age and not given OK-432 during the same period. RESULTS: The administration of OK-432 in most patients was performed on the 4th day after the operation. Pleural drainage could be terminated in a mean of 5.5 days after the intrapleural administration of OK-432. In the control group, the serum C-reactive protein (CRP) level reached a peak on day 4 after the operation and returned to almost a normal level on day 14 after the operation. In the OK-432 group, the peak CRP level, which was significantly higher than that in the control group, was observed on day 7 after the operation and the elevated CRP level was maintained until 28 days after the operation. The mean level of CRP in the OK-432 group was significantly higher than that in the control on days 7, 14, and 28 after the operation. No significant difference was observed in the disease free survivals between the two groups. CONCLUSIONS: Based on the above findings, postoperative prolonged inflammation does not seem to affect the progression of subclinically residual tumor cells. PMID- 11025465 TI - Commentary PMID- 11025464 TI - Identification of the rectal stump after a Hartmann procedure. PMID- 11025466 TI - The role of laparoscopy in the management of gynecologic malignancies. PMID- 11025467 TI - Assessment of nipple discharge. PMID- 11025468 TI - Reply to drs. Leris, kouriefs, williams, and mokbel PMID- 11025469 TI - Clinical use of topoisomerase I inhibitors in anticancer treatment. AB - The camptothecin analogs topotecan and irinotecan have shown to be among the most effective anticancer agents and, as S-phase specific agents, their antitumor effect is maximized when they are administered in protracted schedules. The documented activity as single agents in many adult and pediatric malignancies has been followed by their use in combination with other anticancer agents. These studies have shown promising results, and have placed topotecan and irinotecan in the first line treatment for some malignancies. However, studies to better determine the optimal schedules and sequence of combinations are needed. PMID- 11025470 TI - Hematopoietic stem cell transplantation (HSCT) with a conditioning regimen of busulfan, cyclophosphamide, and etoposide for children with acute myelogenous leukemia (AML): a phase I study of the Pediatric Blood and Marrow Transplant Consortium. AB - BACKGROUND: Hematopoietic stem cell transplantation (HSCT) is an important treatment modality for children with AML. The optimal conditioning regimen is unknown. The aim of this study was to determine the appropriate dosing of etoposide in combination with busulfan and cyclophosphamide in this setting. PROCEDURE: Twenty patients with a diagnosis of AML in first or second remission, or myelodysplasia scheduled for bone marrow transplantation, were included in this study. Patients received busulfan 640 mg/m(2) in 16 doses, cyclophosphamide 120 to 150 mg/kg in two doses, and etoposide from 40-60 mg/kg as a single dose. Extensive toxicity data was collected. RESULTS: Nineteen patients were evaluable for toxicity. Mucositis was seen in all patients. Four patients developed bacteremia and one patient died from overwhelming sepsis on day +3. Four patients developed moderate to severe skin toxicity. The major dose-limiting +3 toxicity was hepatic toxicity, which occurred in 14 of 19 patients. Eight patients developed clinical veno-occlusive disease, including three patients at dose level 4, two of whom had life-threatening disease. This hepatic toxicity defined the MTD of 640 mg/m(2) busulfan, 120 mg/kg of cyclophosphamide, and 60 mg/kg of etoposide. Overall, 9 of 20 patients enrolled in the study survive in remission, 8/14 allogeneic (median follow-up 44 months), and one of six autologous patients (follow-up, 54 months). CONCLUSIONS: We conclude that the combination of busulfan, cyclophosphamide, and etoposide at the doses defined above has activity in the treatment of children with high-risk AML/MDS undergoing allogeneic HSCT. Whether it offers an advantage over other conditioning regimens will require a randomized trial with a larger cohort of patients. PMID- 11025471 TI - Frequency of thiopurine S-methyltransferase genetic variation in Thai children with acute leukemia. AB - BACKGROUND: Thiopurine S-methyltransferase (TPMT) catalyzes the S-methylation (inactivation) of mercaptopurine, azathioprine, and thioguanine, and exhibits genetic variation. About 11% of Caucasians have intermediate TPMT activity because of heterozygosity, and about 1 in 300 inherits TPMT deficiency as an autosomal codominant trait. If patients who have intermediate or deficient TPMT activity receive the standard dose of thiopurine medications, they can accumulate excessive thiopurine nucleotides in hematopoietic tissue, which could lead to severe and possibly fatal myelosuppression. There is very little information about TPMT genetic variation among Asian populations. We investigated the frequency of TPMT genetic variation among Thai children with acute leukemia. PROCEDURE: Fresh whole blood was obtained from 75 Thai children with acute leukemia at the time of remission. Genomic DNA was isolated from total peripheral white blood cells. We performed polymerase chain reaction (PCR) to detect 3 types of variant of the human TPMT gene. RESULTS: Among 75 patients, the frequency of heterozygotes for the TPMT gene among Thai children with acute leukemia was approximately 11%. However, the TPMT*3C was the only variant TPMT allele found among Thai children. This is different from the North American Caucasian populations, in which TPMT*3A is the predominant variant allele, and TPMT*3C is rare (approximately 5% of variant alleles). CONCLUSIONS: There is no difference in the frequency of this genetic variation between Asian and North American Caucasian populations. Determination of the TPMT genotype by PCR method before antileukemic therapy is practical and may have clinical relevance. This knowledge could be applied towards organ transplant recipients who require these medications for immunosuppression. PMID- 11025472 TI - Bone mineral density, body composition, and height in long-term survivors of acute lymphoblastic leukemia in childhood. AB - BACKGROUND: Childhood leukemia has increasing numbers of survivors, so more emphasis is being placed on long-term effects. The ALL-6 protocol of the Dutch Childhood Leukemia Study Group involved high-dose dexamethasone and methotrexate and no cranial irradiation. Therefore, we studied the long-term effects on bone mineral density (BMD), body composition, and growth in survivors of non-high-risk ALL treated with the ALL-6 protocol. PROCEDURE: Twenty-three subjects (12.2-25.4 years) participated in this cross-sectional study. Mean follow-up was 9.6 years (range 7.9-11.4 years). BMD of lumbar spine (LS) and total body (TB) and body composition were measured by dual energy X-ray absorptiometry; results are expressed as standard deviation scores (SDS). Bone mineral apparent density (BMAD(LS)) was calculated to correct for bone size. A questionnaire was administered to determine physical activity, calcium intake, and medical history. RESULTS: Mean SDS for BMD(LS), BMD(TB), and BMAD(LS) were normal. None of the subjects had BMD below -2 SDS; one subject had BMAD(LS) below -2 SDS. Mean SDS for lean body mass, percentage fat, and height were not significantly different from zero. Calcium intake correlated positively with BMD. Nine subjects reported traumatic fractures (eight during or shortly after therapy). CONCLUSIONS: Ten years after ALL-6 treatment, no long-term side effects on height, BMD, or body composition were found in this small group of patients, despite high-dose dexamethasone and methotrexate. This study suggests that ALL treatment without cranial irradiation might not be associated with long-term side effects on growth and BMD. PMID- 11025473 TI - Concurrent Langerhans cell histiocytosis and myelodysplasia in children. AB - BACKGROUND: Langerhans cell histiocytosis (LCH) is characterized by the proliferation of abnormal histiocytes (Langerhans cells), whose origin as a reactive process or a neoplastic disorder is still poorly understood. Although LCH has been recorded as being associated with malignant neoplasms, concurrence of LCH and myelodysplastic syndrome has not been reported so far. PROCEDURE: We report on four children aged 23, 25, 26, and 53 months with multisystem LCH with organ dysfunction (bone marrow and liver) whose bone marrow pictures, taken at diagnosis, revealed the presence of myelodysplastic abnormalities (RA, RAEB, RAEB t). RESULTS: We suggest that the commonly used expression of "organ dysfunction," which refers to clinical and functional alterations, could be explained by a myelodysplastic-like disorder. CONCLUSIONS: The contemporary presence of both events may provide a better understanding of the pathogenesis of LCH, especially in young children with multisystem disease and organ dysfunction, who are known to have a very poor outcome. PMID- 11025474 TI - Is Langerhans cell histiocytosis a myeloid dendritic stem cell disorder related to myelodysplastic disorders? PMID- 11025475 TI - On a new genetic syndrome. PMID- 11025476 TI - Management decisions in children with nephroblastomatosis. PMID- 11025477 TI - Managing nephroblastomatosis PMID- 11025478 TI - Method using urokinase and an antibiotic to avoid device removal in central venous catheter-related infections. PMID- 11025479 TI - Hibernoma in a 13-year-old boy. PMID- 11025480 TI - Multisystem Langerhans-cell histiocytosis with life-threatening pulmonary involvement--good response to cyclosporine A. PMID- 11025481 TI - Management of children with metastatic spinal myxopapillary ependymoma using craniospinal irradiation. PMID- 11025482 TI - Recent developments in quantitative fluorescence calibration for analyzing cells and microarrays. PMID- 11025483 TI - Differential expression of Bax and Bcl-2 in the assessment of cellular dynamics in fine-needle samples of primary breast carcinomas. AB - The rates of cell proliferation and programmed cell death (apoptosis) reflect tumor cell dynamics and are considered to directly influence biological progression and tumor response to therapy. Bax and Bcl-2 are members of a gene family that influence apoptosis and have been used as surrogate markers in the evaluation of this process. Sixty-three fine-needle tumor samples from an equal number of patients with breast carcinomas were analyzed for Bax, Bcl-2, and DNA content by flow cytometry. The results were correlated with classical clinicopathological parameters. Bax values varied widely among tumors and showed no significant correlation with any of the clinicopathological parameters analyzed. Bcl-2 levels ranged from 4% to 91%, correlated positively with estrogen (P = 0.0004) and progesterone (P = 0.0045) receptor positivity, and were more associated with low S-phase tumor values. In contrast, high S-phase values correlated with estrogen receptor negativity, high grade, and DNA aneuploidy. The study results indicate that Bcl-2 and S-phase analysis of fine-needle samples of breast carcinomas provide a convenient tool for the assessment of these tumors. PMID- 11025484 TI - DNA mapping of gastric cancers using flow cytometric analysis. AB - Although numerous studies of gastric cancers on DNA ploidy have been reported, differences in the degree of aneuploidy (DNA index, DI) during progression have not been identified. We attempted to chart the differences in DIs during progression to clarify the role of aneuploidy in gastric cancers. We classified the gastric cancers examined into intestinal (n = 88) and diffuse (n = 48) types, and then analyzed 136 gastric cancers (intramucosal cancer, 42; submucosal cancer, 39; advanced cancer, 55) by flow cytometry using multiple sampling. In addition, we examined the DNA ploidy pattern of mucosal and submucosal lesions using the same submucosal cancers to study the tumor progression in individual cancers. Intratumoral DNA differences in DNA ploidy were observed in both types of gastric cancers. In intestinal-type cancers, multiple subclones indicated by a different DI occurred during the early stage of gastric cancers, whereas in diffuse-type cancers, multiple subclones were found primarily in advanced cancers. Although the DI varied widely in early intestinal-type cancers between 1.0 and 2.0, in early diffuse-type cancers, the DI tended to be less than 1.2. However, in advanced stage gastric cancers, the DI distribution was similar for both histological types. In intestinal-type cancers, high DI (>1.3) aneuploidy was frequently found in mucosal lesions. In contrast, only low DI (<1.2) aneuploid clones were observed in mucosal lesions of diffuse-type cancers. The present results suggest that high DI aneuploid tumor clones in intramucosal cancers acquire invasive ability when they progress to submucosal cancers, whereas DNA aneuploidy itself plays an important role in submucosal invasion of diffuse-type cancers. PMID- 11025485 TI - Immune monitoring in xenotransplantation: the multiparameter flow cytometric mixed lymphocyte culture assay. AB - Xenotransplantation requires monitoring of complex cellular interactions in vitro. A tool to monitor cell proliferation in detail would be instrumental in understanding these cellular interactions in heterogeneous xenogeneic lymphocyte cultures and in patients after xenotransplantation. To accomplish this, we used a fluorescent cell proliferation marker, 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE), in combination with flow cytometry. CFSE, a green fluorescent molecule, binds covalently to intracellular macromolecules. Each cell division reduces the fluorescent intensity per cell by half and shows a characteristic multipeak pattern in flow cytometric analysis. For this study, human lymphocytes were labeled with CFSE and cultured in the presence of irradiated porcine lymphocytes. Cell proliferation was detected in CFSE-labeled lymphocytes in both a single and a multiparameter flow cytometry setting. Concurrently, tritiated ((3)H) thymidine incorporation, a common method to measure gross cell proliferation, was assessed. The kinetics of CFSE-labeled cell proliferation correlated with (3)H-thymidine incorporation in that both methods showed a lag phase for days 1-3 and a log phase for days 4-7. Multiparameter flow cytometric monitoring of mixed lymphocyte cultures allowed phenotyping and assessment of viability of proliferating populations in heterogeneous xenogeneic stimulated human lymphocyte cultures and complemented the classical (3)H thymidine incorporation assay. The use of this technique will allow a wide array of immunologic parameters to be measured in a heterogeneous xenogeneic mixed lymphocyte culture. The information gained from these assays is essential to understanding the biological significance of xenogeneic cellular interaction and for monitoring the immune status of the xenotransplanted patient. PMID- 11025486 TI - Standardized flow cytometric method for the accurate determination of platelet counts in patients with severe thrombocytopenia. AB - BACKGROUND: The therapeutic option of prophylactic platelet (PLT) transfusion in cases of severe thrombocytopenia critically depends on the availability of accurate and precise counts because clinical decisions are widely based on decision or trigger points. Although often applied in current practice at a level of 20 Gpt/L, there is increasing evidence that the trigger points could safely be reduced to 10 or even 5 Gpt/L. In order to facilitate this downward revision, it is necessary to have PLT counting methods that are able to provide reliable results in the appropriate decision range. METHODS: Postchemotherapy-induced pancytopenia PLT counting was performed in patients with hematological malignant disorders. This study describes a novel flow cytometric method that utilizes a PLT-specific monoclonal antibody (CD41a) in conjunction with fluorescent reference beads in order to derive absolute platelet numbers. RESULTS: Applying a mathematical model, this flow cytometric method was shown to have a detection limit of 0.24 Gpt/L and a lower limit of quantification (coefficient of variation [CV] = 10%) of 1.1 Gpt/L. These values are a substantial improvement on previously reported results for the Technicon H1 automated instrument or manual hemocytometry. Moreover, although the flow cytometry and Technicon H3 methods were found by supplementary analyses to show a reasonably good correlation, the hematology instrument showed a distinct tendency to overestimate PLT counts at low levels. CONCLUSION: It is proposed that this standardized immunoplatelet method offers the best approach in evaluating, at the clinical level, the possibility of lower PLT transfusion triggers. It can be used to evaluate the performance limitations of automated hematology analyzers that are widely used at the present time. PMID- 11025487 TI - Alcoholic liver cirrhosis is associated with a decreased expression of the CD28 costimulatory molecule, a lower ability of T cells to bind exogenous IL-2, and increased soluble CD8 levels. AB - Despite the existence of high interleukin (IL)-12 serum levels in patients with chronic active alcoholism, previous studies from our group have shown that, during active ethanol intake, alcoholic patients with alcoholic liver cirrhosis (ALC) display an impaired T-helper-1 response together with abnormalities in the peripheral blood (PB) cytotoxic compartment. The aim of the present study was to gain further insights into the mechanisms underlying these abnormalities. For that purpose, we analyzed the expression on PB B- and T-cell subsets of both the CD28 and CD80 costimulatory molecules, the ability of T lymphocytes to bind to exogenous recombinant IL-2, and the serum levels of soluble CD8 (sCD8) that might interfere with CD8+ T-cell activation in a group of 10 ALC patients with active ethanol intake (ALCET group). As reference groups, we analyzed 10 healthy individuals, 10 chronic alcoholic patients without liver disease (AWLD group) but with active ethanol intake, and 10 ALC patients who had quit drinking for at least 1 year. Our results showed that ALCET patients display a significant decrease in the number of PB CD28+/CD8(hi) T cells (P < 0.05) and CD80+ B cells (P < 0.01) compared with both healthy controls and AWLD patients. In addition, in ALCET patients, PB T cells also showed a decreased ability to bind to exogenous IL-2 (P < 0.01). This was associated with the existence of increased serum levels of sCD8 in ALC patients, the highest levels being detected in the ALCET group (P < 0.01). Altogether, our results point to the existence of several abnormalities that would affect the cytotoxic response in ALCET patients. PMID- 11025489 TI - Forum: journal club PMID- 11025488 TI - Contribution of flow cytometric immunophenotyping to the evaluation of tissues with suspected lymphoma? AB - BACKGROUND: A critical analysis of the contribution of flow cytometric immunophenotyping (FCI) to the evaluation of lymph nodes and extranodal tissues with suspected lymphoma by a large, retrospective approach has not been reported previously and represents the purpose of this study. METHODS: A total of 278 lymph nodes and 95 extranodal tissue specimens submitted over a 2-year period with complete histologic, FCI, and immunohistochemical (IH) data formed the basis of the study. RESULTS: The FCI data contributed significantly to or was consistent with the final tissue diagnosis in the majority (94%) of the tissue samples. There is no well-described utility of flow cytometry markers for Hodgkin's lymphoma (HL) due to the usual scarcity of tumor cells in the final cell suspensions obtained from these tumors. However, the FCI data excluded non Hodgkin's lymphoma (NHL) and suggested the possible usefulness of CD15 and CD30 by FCI in HL. In addition, immunophenotypic data by FCI in combination with touch imprint cytomorphology was useful in excluding a diagnosis of NHL in cases of nonhematopoietic malignancies and was particularly useful in defining the following hematopoietic tumors and malignancies: thymoma, T-cell lymphoblastic lymphoma, leukemia cutis, and plasma cell dyscrasia. Thus, IH was not essential for the diagnosis in these latter cases and was performed in only two cases (one thymoma and one plasma cell dyscrasia). Of interest, FCI supported the diagnosis in 3 cases of Ewing's sarcoma/primitive neuroectodermal tumor by detection of CD56 on the surface of the malignant cell. Only 11% of NHL were "negative" by FCI (i.e., an aberrant T-cell or monoclonal B-cell population was not identified). Reasons for these discrepancies included partial tissue involvement by the NHL with sampling differences, T-cell rich or lymphohistiocytic-rich variants with a small population of monoclonal B cells, marked tumoral sclerosis, poor tumor preservation, and T-cell NHL without an aberrant immunophenotype. Only 60% of CD30+ anaplastic large cell lymphomas (ALCL) were CD30+ by FCI. CONCLUSIONS: FCI data should always be correlated with light microscopy if no FCI abnormalities are detected; IH may need to be performed in selected cases. It is less necessary to perform microscopic examination of tissues when the FCI data are positive and indisputable. However, in selected cases in which FCI data is diagnostic, microscopic observations may provide additional information due to sampling. PMID- 11025490 TI - Patterns and predictors of employer risk-reduction activities (ERRAs) in response to a work-related upper extremity cumulative trauma disorder (UECTD): reports from workers' compensation claimants. AB - BACKGROUND: Despite being preventable, work-related upper extremity cumulative trauma disorders (UECTDs) remain problematic. This study is unique in its focus on predictors of employer risk-reduction activities (ERRAs) in response to a UECTD case. METHODS: Workers' compensation claimants (N = 537) completed a telephone survey about employer risk-reduction activities, workplace characteristics, safety programs, and physician recommendations for job modifications. RESULTS: Only 52% of respondents reported employer actions to investigate or reduce UECTD risk. Engineering and pace changes were prominent for keyboard workers and transfer to another job for manufacturing workers. Safety programs and physician recommendations increased the likelihood of risk-reduction activities. CONCLUSIONS: An opportunity to intervene post-injury to reduce risks for the injured worker and prevent new UECTD cases is being missed. Physician recommendations are strongly associated with specific ERRAs thought to be most effective. Educating employers and physicians about ergonomics could result in prevention of UECTDs. PMID- 11025491 TI - The impact of occupational injury on injured worker and family: outcomes of upper extremity cumulative trauma disorders in Maryland workers. AB - BACKGROUND: Surveys have identified a dramatically rising incidence of work related upper extremity cumulative trauma disorders (UECTDs). Outcome studies have addressed time lost from work and cost of compensation; omitting other significant consequences. We assess health, functional and family outcomes. METHODS: We identified 537 Workers' Compensation UECTD claimants. A computer assisted telephone questionnaire was used to elicit symptom prevalence, functional impairment, depressive symptoms (CES-D scale), employment status. RESULTS: One to 4 years post-claim, respondents reported persistent symptoms severe enough to interfere with work (53%), home/recreation activities (64%) and sleep (44%). Only 64% of responses to the activities of daily living scale items indicated "normal" function. Job loss was reported by 38% of respondents, and depressive symptoms by 31%. CONCLUSIONS: Work-related UECTDs result in persisting symptoms and difficulty in performing simple activities of daily living, impacting home life even more than work. Job loss, symptoms of depression, and family disruption were common. PMID- 11025492 TI - Job stress, upper extremity pain and functional limitations in symptomatic computer users. AB - BACKGROUND: Research suggests that exposure to occupational stressors are related to the presence and/or exacerbation of work-related upper extremity symptoms in office workers. Also worker's response to work demands and/or job stressors (i.e., workstyle) may exacerbate symptom severity and impact function. The present study examines the association among work demands, job stress and workstyle on pain and function. METHODS: 124 symptomatic female office workers completed a questionnaire measuring demographics, medical history, work demands, perception of the work environment, workstyle, pain intensity, functional impact, and time lost from work. RESULTS: Heightened job stress and the tendency to continue to work in a way that contributes to pain to ensure high quality (dimension of workstyle) were related to pain intensity at work and decreased function. These variables, in addition to hours worked per year, were related to increased pain experienced across the work week. The model tested did not predict the occurrence of lost time. CONCLUSIONS: The present findings provide support for the association between job stress, workstyle, upper extremity pain and function. While it is not possible to determine the exact direction of the observed relationships, these results are consistent with prior research indicating the potential significance of job stress and workstyle on symptom exacerbation and functional limitations. Implications for evaluation and intervention are discussed. PMID- 11025493 TI - Risk factors for neck and shoulder disorders: a nested case-control study covering a 24-year period. AB - BACKGROUND: In 1969 a population-based study was conducted in the Stockholm region. From the 2,579 randomly selected participants (18-65 years of age in 1969), the youngest subset were asked to participate in a reexamination in 1993. Information regarding working conditions, conditions outside work, and neck and shoulder disorders was collected retrospectively for the period 1970-1993. METHODS: Of 783 eligible subjects (42-59 years of age in 1993), 484 responded. Cases of neck/shoulder disorders were defined by past sick leave or medical attention or recent symptoms, depending on available information. For each case (n = 271) two controls were randomly selected, matched by age and gender. Variables regarding both physical and psychosocial conditions were included in the matched analyses. RESULTS: Among women mainly psychosocial factors and among men mainly physical factors were associated with neck/shoulder disorders. The only gender common risk indicator found was repetitive hand work (OR approximately 1.5). Interactive effects were also observed. CONCLUSIONS: The impact on neck/shoulder disorders from separate factors was moderate but combinations of physical and psychosocial factors, as well as of work-related and non-work-related factors, produced relative risks above 2. PMID- 11025494 TI - Disability management of low back injuries by employer-retained physicians: ratings and costs. AB - BACKGROUND: Medico-legal models of disability determination for low back pain lack empirical support. Besides diagnostic and functional parameters, social and situational factors may influence impairment/disability ratings and costs. METHODS: Archival data from employer-retained occupational medicine physician files and public court records were examined for 184 Workers' Compensation claimants with low back injuries. Regression was used to predict ratings, costs, and settlement duration from medical, functional, social, and situational variables. RESULTS: Diagnosis, surgery, pain, rating year, and clinic predicted impairment ratings from employer-retained physicians. The clinic effect partially reflected claimant ethnicity. Diagnosis, surgery, tests ordered, legal representation, and impairment rating predicted disability ratings at the administrative law judge level. Diagnosis, tests, and impairment rating predicted costs. For musculoskeletal diagnoses, settlement duration was related negatively to treatment duration and positively to costs. CONCLUSIONS: Social and situational parameters influence disability management among employer-retained physicians, while functional variables have little impact. For musculoskeletal low back pain, increased disability and cost may result from variation in treatment duration. PMID- 11025495 TI - A simple self-rating assessment method of residual work capability for occupational permanent disabilities. AB - BACKGROUND: To explore the validity, reliability, and determinants of a simple self-rating assessment method of residual work capability (RWC) after occupational permanent disabilities. METHODS: Five hundred and thirty-nine compensated permanent disability workers answered three consecutive visual analogue questionnaires wherein they self-rated their residual work capability in terms of speed (RWCS), quality (RWCQ) and a combination of speed and quality (RWCC). At two major hospitals in Taiwan, 169 of these subjects were evaluated with physical capacity assessment (PCA), cognition and sensation assessment (CSA), the work ability index (WAI), and the 12-item Chinese health questionnaire (CHQ-12). RESULTS: High test-retest reliability (Pearson's correlation coefficient 0.77) and satisfactory concurrent validity were shown for RWCS and RWCC. All PCA, CSA and WAI showed significant correlation with RWCs, while CHQ-12 displayed borderline correlation. Employment status after injury and status of the victim's salary as the main source of income for his/her family before injury, were the major determinants of RWCs, in addition to the scales of PCA, CSA and WAI. CONCLUSIONS: The RWCC resulting from the self-rating method may be used as a simple assessment of a victim's residual work capability after occupational permanent disabilities. PMID- 11025496 TI - Flight deck magnetic fields in commercial aircraft. AB - BACKGROUND: Airline pilots are exposed to magnetic fields generated by the aircraft's electrical system. The objectives of this study were (1) to directly measure flight deck magnetic fields in terms of personal exposure to the pilots when flying on different aircraft types over a 75-hour flight-duty month, and (2) to compare magnetic field exposures across flight deck types and job titles. METHODS: Measurements were taken using personal dosimeters carried by either the Captain or the First Officer on Boeing 737/200, Boeing 747/400, Boeing 767/300ER, and Airbus 320 aircraft. RESULTS: Approximately 1,008 block hours were recorded at a sampling frequency of 3 seconds. Total block time exposure to the pilots ranged from a harmonic geometric mean of 6.7 milliGauss (mG) for the Boeing 767/300ER to 12.7 mG for the Boeing 737/200. CONCLUSIONS: Measured flight deck magnetic field levels were substantially above the 0.8-1 mG level typically found in the home or office and suggest the need for further study to evaluate potential health effects of long-term exposure. PMID- 11025497 TI - Petrochemical exposure and menstrual disturbances. AB - BACKGROUND: An exploratory, cross-sectional retrospective study was conducted to examine the effects of benzene exposure on menstrual problems. METHODS: The study was based on a survey administered to over 3,000 women who worked in a large petrochemical company in Beijing, China. An abnormal menstrual cycle length (AMCL), defined as an average menstrual cycle length of greater than 35 days or less than 21 days, is the major outcome of interest. RESULTS: After 7 years of benzene exposure, the adjusted odds ratio of having AMCL for each additional 5 years of exposure was 1.71 (95% CI 1.27-2.31). Feeling stressed at work was also an important predictor. CONCLUSIONS: This study suggests a significant association of benzene exposure and perceived stress with menstrual disturbance. A prospective study is needed to confirm this finding. PMID- 11025498 TI - Mortality from six work-related cancers among African Americans and Latinos. AB - BACKGROUND: Ethnic minorities have been omitted from many occupational health studies, despite their substantial contributions to the labor force and documented cases of high exposures in some settings. To describe the occurrence of potentially work-related cancers among African American and Latino Workers, we conducted an epidemiologic study based on death certificate data. METHODS: Data were obtained from 21 states during 1985-1992. Directly adjusted proportionate mortality ratios (PMRs) and standardized mortality rate ratios (SRRs) for ages 20 64 years were computed for cancers of the lung, nasal cavity, pleura, and peritoneum, malignant melanoma and leukemia using the mortality data and population counts from the 1990 census. RESULTS: Proportionate mortality was generally low for both groups, but African Americans had higher than expected mortality rates for leukemia and cancers of the lung, nasal cavity, and peritoneum. Industry-specific analyses indicate excess leukemia among African American men in the rubber industry (PMR 2.08, 95% confidence interval (CI) 1.29 3.35), Latino men in textile (PMR 2. 31, 95% CI 0.81 - 5.13) and wood industries (PMR 2.03, 95% CI 0.81 - 5.13), and Latino women in the chemical industry (PMR 2.18, 95% CI 0.59 - 8.10), among other findings. Excess cancer of the pleura and peritoneum was observed among workers with a variety of usual occupations, consistent with widespread exposure to asbestos. CONCLUSIONS: This study demonstrates the utility of surveillance data bases for generating basic epidemiologic information on historically neglected workers. Leads about specific workplace exposure can be followed up in more detailed studies. PMID- 11025499 TI - Changes in cellular immunity among workers occupationally exposed to styrene in a plastics lamination plant. AB - BACKGROUND: Styrene is a widely used industrial chemical. Immune and hematological parameters were examined in 29 hand laminators and sprayers exposed to styrene for an average of 14 years and in 19 in-factory unexposed controls. The workers performed hand lamination procedures in a production area with an average area airborne styrene level of 139.5 mg/m(3). Mean concentration of styrene in the blood of exposed workers was 945.7 microg/L and the mean styrene in exhaled air was 38.8 microg/L. METHODS: Parameters of internal and external exposure, immune function assays, immunoglobulins, acute phase reactants and hematology were evaluated in exposed and non-exposed populations. RESULTS: Using multifactorial analysis of variance we found a significant decrease in proliferation of lymphocytes stimulated by Concanavalin A but not by pokeweed mitogen (PWM) in workers occupationally exposed to styrene. Proliferative response to PWM was significantly correlated with the levels of styrene in blood. Phagocytic activity of monocytes, levels of IgG, IgA, IgM, IgE and alpha-2 macroglobulin in serum were indistinguishable in the two groups. The population exposed to styrene had increased levels of C4-component of complement. Levels of C3-component of complement were positively correlated with duration of exposure. A significant elevation in the percentage and number of monocytes and a significantly decreased number of lymphocytes were seen in exposed workers. Styrene concentrations in both blood and exhaled air were associated with decreased percentage of large granular lymphocytes. CONCLUSIONS: These results suggest immune alterations of cell-mediated immune response of T-lymphocytes and imbalance in leucocyte subsets in peripheral blood of workers exposed to styrene. PMID- 11025500 TI - Empowerment evaluation of worker safety and health education programs. AB - BACKGROUND: The need to expand the use of evaluation to learn how education programs contribute to workers' occupational safety and health is well documented. Similarly recognized is the need to expand workers' involvement as primary stakeholders in program evaluation. METHODS: Articles for this review were identified through computer database and manual searches related to: intervention research and evaluation; occupational safety, health training, and education; and worker participation and empowerment. RESULTS: After identifying empowerment as a multilevel and multidimensional concept, this review used a theoretical framework of evaluation to show how various participatory and empowering approaches can affect evaluation studies and their use. CONCLUSIONS: The field of occupational safety and health has a unique historical opportunity to further expand workers' involvement in their own education through the use of participatory and empowering approaches to evaluation. Use of these approaches has the potential to strengthen capacities for organizational learning and improve both program theory and practice. PMID- 11025503 TI - Editorial PMID- 11025501 TI - Industrial emergency response training: an assessment of long-term impact of a union-based program. AB - BACKGROUND: The long-term impact of the United Automobile Workers' (UAW) Industrial Emergency Response Training on health and safety conditions and practices in plants is described. Two strategies are combined in this 24-hour training: the use of peer (worker) trainers and a participant-centered approach, the Small Group Activity Method (SGAM). METHODS: Impact was assessed through a telephone survey of 67 trainees conducted on average 16.8 months post-training. RESULTS: 74% of trainees reported using at least one of the provided written training materials after training. Of those reporting an accident in their facility after training, 88% said it was handled differently because of training. Trainees report that peer-trainers are more knowledgeable and communicate better than do other types of trainers. Trainees also reported that SGAM made training more applicable to their workplace. 98.5% of trainees like the methods utilized. CONCLUSIONS: Worker-trainers and SGAM are effective training methods enhancing the long-term impact of union-based workplace health and safety training programs. PMID- 11025504 TI - Turbo ASL: arterial spin labeling with higher SNR and temporal resolution. AB - A modified pulsed arterial spin labeling (ASL) technique is introduced here that has both higher temporal resolution and higher SNR per unit time than existing ASL techniques. In this technique, the time TI between the application of the tag and image acquisition is longer than the repetition time TR, allowing for the use of greatly reduced TR values without a significant decrease in the amplitude of the ASL signal. This improves both the temporal resolution and the sensitivity of ASL for functional brain mapping. PMID- 11025505 TI - Gallbladder localization of (19)F MRS catabolite signals in patients receiving bolus and protracted venous infusional 5-fluorouracil. AB - The hepatobiliary distribution of 5-fluorouracil (5FU) catabolites was investigated in nine patients. Using fluorine 3D-chemical shift imaging, four patients receiving protracted venous infusion of 5FU demonstrated catabolite localized to the gallbladder. No hepatobiliary fluorine signals were detected in three patients whose gallbladders were absent or abnormal. Signals from the gallbladder showed a 2.2-2.4 ppm high-frequency shift from alpha-fluoro-beta alanine, suggesting the presence of alpha-fluoro-beta-alanine-bile-acid conjugates. 3D-chemical shift imaging of two patients receiving bolus 5FU revealed alpha-fluoro-beta-alanine to be localized to the liver within 1 hr of administration. In one patient examined 4 hr after bolus administration, catabolite signal was detected only in the gallbladder. PMID- 11025506 TI - Line scan echo planar spectroscopic imaging. AB - A line scan echo planar spectroscopic imaging (LSEPSI) sequence is presented which can rapidly produce 2D chemical shift imaging (CSI) data with minimal relaxation weighting and motion-related artifacts. The technique is based on successive "snapshot" 1D CSI acquisitions of individual tissue columns, and avoids T(1) saturation problems associated with the short TR periods needed for very rapid scanning with either conventional or echo planar-based 2D CSI methods. Potential applications include rapid fat/water spectral quantitation in the abdomen and internally referenced temperature monitoring for interventional procedures. PMID- 11025507 TI - Three-dimensional tailored RF pulses for the reduction of susceptibility artifacts in T(*)(2)-weighted functional MRI. AB - A three-dimensional tailored RF pulse method for reducing intravoxel dephasing artifacts in T *(2)-weighted functional MRI is presented. A stack of spirals k space trajectory is employed to excite a disk of magnetization for small tip angles. Smaller disks with a linear through-plane phase are inserted into the disk to locally refocus regions which are normally dephased due to susceptibility variations. Numerical simulations and imaging experiments which use the tailored RF pulses are presented. Limitations of the method and improvements are also discussed. PMID- 11025508 TI - Gradient-induced acoustic and magnetic field fluctuations in a 4T whole-body MR imager. AB - Both the acoustic and magnetic fluctuation frequency response functions for a Siemens AS25 body gradient coil inside a 4 Tesla whole-body MR system were measured and analyzed in this study. In an attempt to correlate the acoustic noise inside the gradient coil with magnetic field oscillations, triangular and trapezoidal gradient impulses of varying amplitudes and widths were used to excite the gradient coil. The acoustic and magnetic responses to these inputs were measured. The results show the existence of discrete resonances in both acoustic and uniform magnetic field fluctuation spectra, while gradient magnetic field fluctuation spectra show no such resonances. In addition, the dominant amplitude peaks in spectra fluctuate similarly with respect to trapezoidal gradient impulse flat-top widths. This implies that these phenomena are correlated, and that the trapezoidal impulse flat-top width may be used as a way to suppress both acoustic noise and uniform magnetic field oscillations. PMID- 11025509 TI - Confidence images for MR spectroscopic imaging. AB - Automated spectral analysis and estimation of signal amplitudes from magnetic resonance data generally constitutes a difficult nonlinear optimization problem. Obtaining a measure of the degree of confidence that one has in the estimated parameters is as important as the estimates themselves. This is particularly important if clinical diagnoses are to be based on estimated metabolite levels, as in applications of MR Spectroscopic Imaging for human studies. In this report, a standard method of obtaining confidence intervals for nonlinear estimation is applied to simulated data and short-TE clinical proton spectroscopic imaging data sets of human brain. So-called "confidence images" are generated to serve as visual indicators of how much trust should be placed in interpretation of spatial variations seen in images derived from fitted metabolite parameter estimates. This method is introduced in a Bayesian framework to enable comparison with similar techniques using Cramer-Rao bounds and the residuals of fitted results. PMID- 11025510 TI - Extended ISIS sequences insensitive to T(1) smearing. AB - Image selected in vivo spectroscopy (ISIS) is a volume selection method often used for in vivo (31)P MRS, since it is suitable for measurements of substances with short T(2). However, ISIS can suffer from significant signal contributions caused by T(1) smearing from regions outside the VOI. A computer model was developed to simulate this contamination. The simulation results for the ISIS experiment order implemented in our MR system (ISIS-0) were in agreement with results obtained from phantom measurements. A new extended ISIS experiment order (E-ISIS) was developed, consisting of four "optimal" ISIS experiment orders (ISIS 1 to ISIS-4) performed consecutively with dummy ISIS experiments in between. The simulation results show that contamination due to T(1) smearing is, effectively, eliminated with E-ISIS and is significantly lower than for ISIS-0 and ISIS-1. E ISIS offers increased accuracy for quantitative and qualitative determination of substances studied using in vivo MRS. Hence, E-ISIS can be valuable for both clinical and research applications. PMID- 11025511 TI - Simultaneous in vivo monitoring of hepatic glucose and glucose-6-phosphate by (13)C-NMR spectroscopy. AB - Hepatic glucose-6-phosphate (G6P) was monitored non-invasively in rat liver by in vivo (13)C NMR spectroscopy after infusion of [1-(13)C] glucose. The phosphorylation of glucose to G6P yields small but characteristic displacements for all of its (13)C-NMR resonances relative to those of glucose. It is demonstrated that in vivo (13)C-NMR spectroscopy at 7 Tesla provides the spectral sensitivity and resolution to detect hepatic G6P present at sub-millimolar concentration as partially resolved low-field shoulders of the glucose C1 resonances at 96.86 ppm (C1beta) and 93. 02 ppm (C1alpha). Upon (13)C-labeling, the intracellular conversion of [1-(13)C] glucose to [1-(13)C] G6P could be monitored, which allowed the hepatic glucose-G6P substrate cycle to be assessed in situ. The close correlation found for the (13)C labeling patterns of glucose and G6P supports the concept of an active substrate cycle whose rate exceeds that of net hepatic glucose metabolism. High-resolution (13)C-NMR spectroscopy and biochemical analyses of tissue biopsies collected at the end of the experiments confirmed qualitatively the findings obtained in vivo. PMID- 11025512 TI - Determination of the MRI contrast agent concentration time course in vivo following bolus injection: effect of equilibrium transcytolemmal water exchange. AB - For bolus-tracking studies, it is commonly assumed that CR concentration bears a linear relationship with the measured (usually longitudinal) (1)H(2)O relaxation rate constant, R*(1) identical with(T(1) *)(-1). This requires that equilibrium transcytolemmal water exchange be in the fast exchange limit (FXL). However, though systems remain in fast exchange, the FXL will not usually obtain. Here, the consequences are considered: 1) the measurement of R(1) * itself can be affected, 2) the resultant non-linear [CR]-dependence causes significant error by assuming FXL, 3) the thermodynamic [CR] (based on the space in which CR is actually distributed) can be determined, 4) transcytolemmal water permeability may be estimated, and 5) the pharmacokinetic parameters can be factored. For a 30 sec, 0.17 mmol/kg dose of GdDTPA(2-), the FXL assumption underestimates the [CR] maximum in rat thigh muscle by a factor of almost two. Similar results are obtained for a rat brain GS-9L gliosarcoma tumor model. PMID- 11025513 TI - Selective contrast-enhanced MR angiography. AB - In this study the feasibility of intraarterial contrast administration was investigated. Its use for navigation and treatment evaluation during MR-guided intravascular interventions was explored in phantom and animal experiments. An injection protocol was developed, which accounts for sequence parameters and vessel flow rate. Tracking a bolus of contrast agent was useful to verify the catheter tip position and to assess flow conditions. Compared to intravenous contrast-enhanced magnetic resonance angiography (CE-MRA), selective contrast administration permitted a strongly reduced dose. In two-dimensional (2D) acquisitions overlap of vessels was prevented. Injection and acquisition were easily and accurately synchronized in selective 3D CE-MRA, and a high contrast concentration could be maintained during the entire acquisition. Selective injection is useful in the course of an intervention, to facilitate navigation, provide information on flow conditions, and to evaluate treatment progress repeatedly. PMID- 11025514 TI - Magnetic resonance diffusion tensor imaging for characterizing diffuse and focal white matter abnormalities in multiple sclerosis. AB - High-resolution diffusion tensor imaging (DTI) was performed in 14 patients with clinically definite multiple sclerosis (MS) and the trace of the diffusion tensor () and the fractional anisotropy (FA) were determined in normal appearing white matter (NAWM) and in different types of focal MS lesions. A small but significant increase of the in NAWM compared to control white matter ((840 +/ 85) x 10(-6) mm(2)/sec vs. (812 +/- 59) x 10(-6) mm(2)/sec; P < 0.01) was found. In addition, there was a significant decrease in the FA of normal-appearing regions containing well-defined white matter tracts, such as the genu of the internal capsule. In non-acute lesions, the of T(1)-hypointense areas was significantly higher than that of T(1)-isointense lesions ((1198 +/- 248) x 10( 6) mm(2)/sec vs. (1006 +/- 142) x 10(-6) mm(2)/sec; P < 0. 001), and there was a corresponding inverse relation of FA. Diffusion characteristics of active lesions with different enhancement patterns were also significantly different. DTI with a phase navigated interleaved echo planar imaging technique may be used to detect abnormalities of isotropic and anisotropic diffusion in the NAWM and selected fiber tracts of patients with MS throughout the entire brain, and it demonstrates substantial differences between various types of focal lesions. PMID- 11025515 TI - Elastic registration of 3D cartilage surfaces from MR image data for detecting local changes in cartilage thickness. AB - The objective of this work was to develop and validate a computational method for the registration (matching) of 3D cartilage plates from MR image data sets. The technique tracks local cartilage thickness changes over time. A 3D elastic registration technique was applied that identifies corresponding points of the bone-cartilage interface in MR data sets of 3D-reconstructed cartilage plates. In a first rigid preregistration step, the surfaces are aligned, using the principal axes decomposition to correct for different joint positions and orientations in the MR scanner. In a second step, the surfaces are deformed elastically, based on geometric surface features, until they are sufficiently similar to identify corresponding surface points. The method was validated against artificially corrupted cartilage surfaces and MR data obtained from in vivo and in vitro compression experiments. The in vivo reproducibility was tested on patellar data sets of volunteers, with repositioning of the joint in between replicate acquisitions. PMID- 11025516 TI - Partially parallel imaging with localized sensitivities (PILS). AB - In this study a novel partially parallel acquisition method is presented, which can be used to accelerate image acquisition using an RF coil array for spatial encoding. In this technique, Parallel Imaging with Localized Sensitivities (PILS), it is assumed that the individual coils in the array have localized sensitivity patterns, in that their sensitivity is restricted to a finite region of space. Within the PILS model, a detailed, highly accurate RF field map is not needed prior to reconstruction. In PILS, each coil in the array is fully characterized by only two parameters: the center of coil's sensitive region in the FOV and the width of the sensitive region around this center. In this study, it is demonstrated that the incorporation of these coil parameters into a localized Fourier transform allows reconstruction of full FOV images in each of the component coils from data sets acquired with a reduced number of phase encoding steps compared to conventional imaging techniques. After the introduction of the PILS technique, primary focus is given to issues related to the practical implementation of PILS, including coil parameter determination and the SNR and artifact power in the resulting images. Finally, in vivo PILS images are shown which demonstrate the utility of the technique. PMID- 11025517 TI - Specific absorption rate study for radiofrequency current density imaging using a two-dimensional finite element model. AB - Radiofrequency current density imaging is an MR technique that images tissue conductivity contrast. Compared to conventional MRI, RF-CDI uses two additional sources of RF power to be absorbed and that must be evaluated in terms of proper parameter optimization to prevent excessive tissue heating and effects on the nervous system. In view of possible future clinical use of RF-CDI, a simple 2D finite element model of a rat brain was built to simulate current density distribution and distribution of absorbed RF power, i.e., SAR and related tissue heating. Current density in the rat brain was also evaluated qualitatively and quantitatively in an in vivo RF-CDI experiment. The results demonstrate that a numerical model can predict SAR and tissue temperature changes. The study also shows that substantial sensitivity and resolution of RF-CDI can be achieved using imaging parameters that produce SAR and temperature changes within allowed limits. PMID- 11025518 TI - Gradient preemphasis calibration in diffusion-weighted echo-planar imaging. AB - This article describes a method which enables fast and objective pulse-sequence specific preemphasis calibration, using standard pulse sequences and system hardware. The method is based on a k-space measurement technique, and has been applied to single-shot, diffusion-weighted, spin-echo, echo-planar imaging (DW-SE EPI), which is particularly sensitive to eddy-current-induced image distortions. The efficiency of the technique was demonstrated not only by the reduction of eddy-current fields to a negligible level using full preemphasis compensation, but also by the fact that adjustment of the slow time-base alone sufficed for the practical elimination of image distortions in the DW-SE-EPI images and the subsequent diffusion tensor maps (in a phantom and a human brain). By seeking to eliminate directly the effect of eddy-current-induced phase shifts during the EPI data collection, the method is free of the complications and restrictions associated with other eddy-current correction techniques for DW-SE-EPI (such as acquisition of additional calibration scans, intense postprocessing, extensive pulse-sequence modifications), making their use redundant. PMID- 11025519 TI - In vivo fiber tractography using DT-MRI data. AB - Fiber tract trajectories in coherently organized brain white matter pathways were computed from in vivo diffusion tensor magnetic resonance imaging (DT-MRI) data. First, a continuous diffusion tensor field is constructed from this discrete, noisy, measured DT-MRI data. Then a Frenet equation, describing the evolution of a fiber tract, was solved. This approach was validated using synthesized, noisy DT-MRI data. Corpus callosum and pyramidal tract trajectories were constructed and found to be consistent with known anatomy. The method's reliability, however, degrades where the distribution of fiber tract directions is nonuniform. Moreover, background noise in diffusion-weighted MRIs can cause a computed trajectory to hop from tract to tract. Still, this method can provide quantitative information with which to visualize and study connectivity and continuity of neural pathways in the central and peripheral nervous systems in vivo, and holds promise for elucidating architectural features in other fibrous tissues and ordered media. PMID- 11025520 TI - Novel quadrature birdcage coil for a vertical B(0) field open MRI system. AB - A birdcage coil possesses various intrinsic resonant modes. One of the modes produces sinusoidal current distribution around the rung elements of the birdcage and generates a highly uniform B(1) field in the transverse plane. Another mode is the end-ring resonant mode, in which there are no currents in the rungs of the birdcage, leaving the currents flowing in the top and bottom end-rings. Thus, this mode behaves like a "Helmholtz pair," producing a highly uniform B(1) field along the axis of the birdcage. By combining the sinusoidal resonant mode and the end-ring resonant mode at the same frequency, a quadrature birdcage coil for a vertical B(0) field is made possible. In particular, the coil dimension can be chosen such that the two end-ring loops behave as a Helmholtz pair, assuring the optimized B(1) field uniformity around the coil center region. The coil is well suited for a vertical B(0) MRI system, as both B(1) fields are perpendicular to the vertical B(0) field. To mimic a vertical B(0) field orientation, an existing horizontal bore MRI system was used with the RF coil oriented to simulate a "vertical" B(0) field. This is a report of the successful quadrature implementation of this novel birdcage coil. PMID- 11025521 TI - Spectroscopic lineshape correction by QUECC: combined QUALITY deconvolution and eddy current correction. AB - Lineshape distortion due to residual eddy currents and magnetic field inhomogeneities are often present in short echo time (1)H spectroscopic data. Lineshape correction methods such as QUALITY deconvolution and eddy current correction (ECC), which use a separate reference spectrum for lineshape correction, have shortcomings when unsuppressed water is chosen as the reference. This paper outlines a method of integrating both techniques to overcome these limitations while still using unsuppressed water as the reference signal. This hybrid lineshape correction technique (QUECC) is demonstrated in vivo using stimulated echo acquisition mode (STEAM) localized 4.0 Tesla data. Metabolite quantification precision increased by an average of 7%-46% compared to QUALITY deconvolution (depending on filtering) and by an average of 6% compared to ECC. PMID- 11025522 TI - Choice of spectroscopic lineshape model affects metabolite peak areas and area ratios. AB - The use of Lorentzian model lineshapes leads to systematic errors in the quantification of in vivo (1)H NMR spectra. Experimental lineshapes are better modeled by the Voigt (mixed Lorentzian-Gaussian) function, leading to more accurate fits (reduced chi(2)). In this work, results from a group of 41 subjects are presented. It is shown that not only are the estimated metabolite peak areas affected by the choice of lineshape model, but so too are the metabolite ratios. For example, the NAA/choline ratio was 1.92 +/- 0.06 (mean +/- standard error) using the Lorentzian lineshape model and 1.85 +/- 0.05 using the Voigt lineshape model. The corresponding figures for NAA/creatine were 2.32 +/- 0.06 and 2. 10 +/ 0.05 respectively, which are significantly different for the two lineshape models. An explanation of this previously unreported effect is given. This finding clearly has serious implications for the methodology and reporting of spectroscopic studies. PMID- 11025523 TI - Monitoring water content in deforming intervertebral disc tissue by finite element analysis of MRI data. AB - Mechanical loading, occurring during normal daily life, causes fluid to be expelled from intervertebral discs. Excessive fluid loss during heavy loading might make the disc more vulnerable to damage. In this study, fluid loss was investigated in vitro through monitoring the loss of MRI signal intensity in four bovine coccygeal intervertebral discs, compressed at 2000 N during 1.5 hr. The MRI signals were analyzed with the aid of finite element models to account for the deformation of the tissue. A gradual signal loss over time was found during loading, the most pronounced loss occurring in the central disc region. Initial patterns of signal distribution were quite variable between specimens but repeatable within specimens. PMID- 11025525 TI - Simultaneous image acquisition utilizing hybrid body and phased array receiver coils. AB - In clinical MR imaging the design and selection of receiver coil is an important step in ensuring the highest image quality. Often this choice is based on selecting a receiver coil characterized by high spatial uniformity such as the body and head volume receiver coils or a surface coil (or array of coils) that provide high signal-to-noise ratio (SNR). In the past, it has been difficult to accomplish both high SNR and spatial uniformity as both coil types achieve one of these characteristics at the expense of the other. The purpose of this study was to achieve both high SNR and spatial uniformity through the simultaneous acquisition of the MR signal using the body and a surface coil array. Results indicate that this hybrid system can provide uniformity and SNR values comparable to those achieved by the body and surface coil arrays, respectively. PMID- 11025524 TI - Contrast agent-enhanced magnetic resonance imaging of skeletal muscle damage in animal models of muscular dystrophy. AB - Membrane lesions play an early role in the pathogenesis of muscular dystrophy. Using a new albumin-targeted contrast agent (MS-325), sarcolemmal integrity of two animal models for muscular dystrophy was studied by MRI. Intravenously injected MS-325 does not enter skeletal muscle of normal mice. However, mdx and Sgca-null mutant mice, animal models for Duchenne and sarcoglycan-deficient limb girdle muscular dystrophy, respectively, showed significant accumulation of MS 325 in skeletal muscle. The results suggest that contrast agent-enhanced MRI could serve as a common, noninvasive imaging procedure for evaluating the localization, extent, and mechanisms of skeletal muscle damage in muscular dystrophy. Furthermore, this method is expected to facilitate assessment of therapeutic approaches in these diseases. PMID- 11025528 TI - The dual origin of the yeast mitochondrial proteome. AB - We propose a scheme for the origin of mitochondria based on phylogenetic reconstructions with more than 400 yeast nuclear genes that encode mitochondrial proteins. Half of the yeast mitochondrial proteins have no discernable bacterial homologues, while one-tenth are unequivocally of alpha-proteobacterial origin. These data suggest that the majority of genes encoding yeast mitochondrial proteins are descendants of two different genomic lineages that have evolved in different modes. First, the ancestral free-living alpha-proteobacterium evolved into an endosymbiont of an anaerobic host. Most of the ancestral bacterial genes were lost, but a small fraction of genes supporting bioenergetic and translational processes were retained and eventually transferred to what became the host nuclear genome. In a second, parallel mode, a larger number of novel mitochondrial genes were recruited from the nuclear genome to complement the remaining genes from the bacterial ancestor. These eukaryotic genes, which are primarily involved in transport and regulatory functions, transformed the endosymbiont into an ATP-exporting organelle. PMID- 11025529 TI - Predicting the function and subcellular location of Caenorhabditis elegans proteins similar to Saccharomyces cerevisiae beta-oxidation enzymes. AB - The role of peroxisomal processes in the maintenance of neurons has not been thoroughly investigated. We propose using Caenorhabditis elegans as a model organism for studying the molecular basis underlying neurodegeneration in certain human peroxisomal disorders, e.g. Zellweger syndrome, since the nematode neural network is well characterized and relatively simple in function. Here we have identified C. elegans PEX-5 (C34C6.6) representing the receptor for peroxisomal targeting signal type 1 (PTS1), defective in patients with such disorders. PEX-5 interacted strongly in a two-hybrid assay with Gal4p-SKL, and a screen using PEX 5 identified interaction partners that were predominantly terminated with PTS1 or its variants. A list of C. elegans proteins with similarities to well characterized yeast beta-oxidation enzymes was compiled by homology probing. The possible subcellular localization of these orthologues was predicted using an algorithm based on trafficking signals. Examining the C termini of selected nematode proteins for PTS1 function substantiated predictions made regarding the proteins' peroxisomal location. It is concluded that the eukaryotic PEX5 dependent route for importing PTS1-containing proteins into peroxisomes is conserved in nematodes. C. elegans might emerge as an attractive model system for studying the importance of peroxisomes and affiliated processes in neurodegeneration, and also for studying a beta-oxidation process that is potentially compartmentalized in both mitochondria and peroxisomes. PMID- 11025530 TI - Global cDNA amplification combined with real-time RT-PCR: accurate quantification of multiple human potassium channel genes at the single cell level. AB - We have developed a sensitive quantitative RT-PCR procedure suitable for the analysis of small samples, including single cells, and have used it to measure levels of potassium channel mRNAs in a panel of human tissues and small numbers of cells grown in culture. The method involves an initial global amplification of cDNA derived from all added polyadenylated mRNA followed by quantitative RT-PCR of individual genes using specific primers. In order to facilitate rapid and accurate processing of samples, we have adapted the approach to allow use of TaqMan real-time quantitative PCR. We demonstrate that the approach represents a major improvement over existing conventional and real-time quantitative PCR approaches, since it can be applied to samples equivalent to a single cell, is able to accurately measure expression levels equivalent to less than 1/100th copy/cell (one specific cDNA molecule present amongst 10(8) total cDNA molecules). Furthermore, since the initial step involves a global amplification of all expressed genes, a permanent cDNA archive is generated from each sample, which can be regenerated indefinitely for further expression analysis. PMID- 11025531 TI - Expression profiling of single mammalian cells--small is beautiful. AB - Increasingly mRNA expression patterns established using a variety of molecular technologies such as cDNA microarrays, SAGE and cDNA display are being used to identify potential regulatory genes and as a means of providing valuable insights into the biological status of the starting sample. Until recently, the application of these techniques has been limited to mRNA isolated from millions or, at very best, several thousand cells thereby restricting the study of small samples and complex tissues. To overcome this limitation a variety of amplification approaches have been developed which are capable of broadly evaluating mRNA expression patterns in single cells. This review will describe approaches that have been employed to examine global gene expression patterns either in small numbers of cells or, wherever possible, in actual isolated single cells. The first half of the review will summarize the technical aspects of methods developed for single-cell analysis and the latter half of the review will describe the areas of biological research that have benefited from single-cell expression analysis. PMID- 11025532 TI - The gene guessing game. AB - A recent flurry of publications and media attention has revived interest in the question of how many genes exist in the human genome. Here, I review the estimates and use genomic sequence data from human chromosomes 21 and 22 to establish my own prediction. PMID- 11025533 TI - Featured organism: Danio rerio, the zebrafish. AB - The zebrafish has long been a favourite model for the study of vertebrate development. Here we provide an overview of the current state of knowledge and resources for the study of this fish, with comments on the future direction of zebrafish genomics from Professor Mark Fishman and Dr Stephen Wilson. PMID- 11025534 TI - Zebrafish functional genomics. Interview by Joanne Wixon. PMID- 11025535 TI - Functional genomics at the Arabidopsis meeting. PMID- 11025536 TI - The second international meeting on Microarray Data Standards, Annotations, Ontologies and Databases. PMID- 11025537 TI - Life will never be the same. Annual Genome Sequencing and Biology Meeting, Cold Spring Harbor Laboratory, USA. May 2000. PMID- 11025538 TI - Website review: UK CropNet. AB - This review explores the UK CropNet site. The project is aimed at aiding the comparative mapping of cereal and other crop genomes. The site provides software tools for use by those working on genome mapping, and access to an array of databases that will be of interest to all members of the plant genomics research community, using several ACeDB interfaces. All screen views from the website are reproduced with the kind permission of Dr Sean May, Director, Nottingham Arabidopsis Stock Centre (NASC). PMID- 11025540 TI - Bayesian probabilistic approach for predicting backbone structures in terms of protein blocks. AB - By using an unsupervised cluster analyzer, we have identified a local structural alphabet composed of 16 folding patterns of five consecutive C(alpha) ("protein blocks"). The dependence that exists between successive blocks is explicitly taken into account. A Bayesian approach based on the relation protein block-amino acid propensity is used for prediction and leads to a success rate close to 35%. Sharing sequence windows associated with certain blocks into "sequence families" improves the prediction accuracy by 6%. This prediction accuracy exceeds 75% when keeping the first four predicted protein blocks at each site of the protein. In addition, two different strategies are proposed: the first one defines the number of protein blocks in each site needed for respecting a user-fixed prediction accuracy, and alternatively, the second one defines the different protein sites to be predicted with a user-fixed number of blocks and a chosen accuracy. This last strategy applied to the ubiquitin conjugating enzyme (alpha/beta protein) shows that 91% of the sites may be predicted with a prediction accuracy larger than 77% considering only three blocks per site. The prediction strategies proposed improve our knowledge about sequence-structure dependence and should be very useful in ab initio protein modelling. PMID- 11025541 TI - Mechanism of protein folding. AB - The high structural resolution of the main transition states for the formation of native structure for the six small proteins of which Phi-values for a large set of mutants have become available, barstar, barnase, chymotrypsin inhibitor 2, Arc repressor, the src SH3 domain, and a tetrameric p53 domain reveals that for the first 5 of these proteins: (1) Residues that belong to regular secondary structure have a significantly larger average fraction of native structural consolidation than residues in loops; (2) on the other hand, secondary and tertiary structures have built up to the same degree, or at least a high degree, but nonuniformly distributed over the molecule; (3) the most consolidated parts of each protein molecule in the transition state cluster together, and these clusters contain a significantly higher percentage of residues that belong to regular secondary structure than the rest of the molecule. These observations further reconcile the framework model with the nucleation-condensation mechanism for folding: The amazing speed of protein folding can be understood as caused by the catalytic effect of the formation of clusters of residues which have particularly high preferences for the early formation of regular secondary structure in the presence of significant amounts of tertiary structure interactions. PMID- 11025539 TI - Current awareness on comparative and functional genomics. PMID- 11025542 TI - Molecular-dynamics simulation of the beta domain of metallothionein with a semi empirical treatment of the metal core. AB - The three-metal-containing beta domain of rat liver metallothionein-2 in aqueous solution was simulated with different metal contents. The Cd(3), the CdZn(2), and the Zn(3) variant were investigated using a conventional molecular dynamics simulation, as well as a simulation with a semi-empirical quantum-chemical description (MNDO and MNDO/d) of the metal core embedded in a classical environment. For the purely classical simulations, the standard GROMOS96 force field parameters were used, and parameters were estimated for cadmium. The results of both kinds of simulations were compared to each other and to the corresponding experimental X-ray crystallographic and NMR solution data. The purely classical simulations were found to produce a too compact metal cluster with partially incorrect geometries, which affected the enfolding protein backbone structure. The inclusion of MNDO/d for the treatment of the metal cluster improved the results to give correct cluster geometries and an overall protein structure in agreement with the experiment. The metal cluster and the cysteine residues bound to it are structurally stable, while the irregular polypeptide backbone loops between the cysteines exhibit a considerable flexibility. MNDO without extension to d orbitals failed to maintain the structure of the metal core. PMID- 11025543 TI - Alpha-amylase inhibitors selected from a combinatorial library of a cellulose binding domain scaffold. AB - A disulfide bridge-constrained cellulose binding domain (CBD(WT)) derived from the cellobiohydrolase Cel7A from Trichoderma reesei has been investigated for use in scaffold engineering to obtain novel binding proteins. The gene encoding the wild-type 36 aa CBD(WT) domain was first inserted into a phagemid vector and shown to be functionally displayed on M13 filamentous phage as a protein III fusion protein with retained cellulose binding activity. A combinatorial library comprising 46 million variants of the CBD domain was constructed through randomization of 11 positions located at the domain surface and distributed over three separate beta-sheets of the domain. Using the enzyme porcine alpha-amylase (PPA) as target in biopannings, two CBD variants showing selective binding to the enzyme were characterized. Reduction and iodoacetamide blocking of cysteine residues in selected CBD variants resulted in a loss of binding activity, indicating a conformation dependent binding. Interestingly, further studies showed that the selected CBD variants were capable of competing with the binding of the amylase inhibitor acarbose to the enzyme. In addition, the enzyme activity could be partially inhibited by addition of soluble protein, suggesting that the selected CBD variants bind to the active site of the enzyme. PMID- 11025544 TI - Structural consequences of an amino acid deletion in the B1 domain of protein G. AB - We describe the NMR structure of a deletion mutant of the B1 IgG-binding domain from Group G Streptococcus. The deletion occurs within the last beta-strand of the protein, where it may potentially have a deleterious effect on the stability of the protein if the protein were not able to conformationally adjust to the perturbation. In particular, the deletion changes the registry of the final three residues in the sheet, forcing a polar Thr to be buried in the interior of the protein and exposing a hydrophobic Val to solvent. The deletion could also potentially create a large cavity in the beta-sheet and force the alpha- and gamma-carboxylates of the C-terminal Glu residue into a partially buried region of the sheet. The structure of the mutant illustrates how the conformation of the protein adjusts to the deletion, thereby mitigating some of the potentially deleterious consequences. Although the elements of secondary structure are retained between the mutant and the wt domain, there are multiple small adjustments in the segments connecting secondary structure elements. In particular, a hydrogen bond between the Glu57 carboxylates and two main chain amides is introduced that alters the conformation in the loop connecting the helix to strand 3. In addition, to minimize hydrophobic surface exposure, the turn connecting strands 1 and 2 folds toward the core so that the molecular volume is decreased. PMID- 11025545 TI - NMR structures of thioredoxin m from the green alga Chlamydomonas reinhardtii. AB - Chloroplast thioredoxin m from the green alga Chlamydomomas reinhardtii is very efficiently reduced in vitro and in vivo in the presence of photoreduced ferredoxin and a ferredoxin dependent ferredoxin-thioredoxin reductase. Once reduced, thioredoxin m has the capability to quickly activate the NADP malate dehydrogenase (EC 1.1.1.82) a regulatory enzyme involved in an energy-dependent assimilation of carbon dioxide in C4 plants. This activation is the result of the reduction of two disulfide bridges by thioredoxin m, that are located at the N- and C-terminii of the NADP malate dehydrogenase. The molecular structure of thioredoxin m was solved using NMR and compared to other known thioredoxins. Thioredoxin m belongs to the prokaryotic type of thioredoxin, which is divergent from the eukaryotic-type thioredoxins also represented in plants by the h (cytosolic) and f (chloroplastic) types of thioredoxins. The dynamics of the molecule have been assessed using (15)N relaxation data and are found to correlate well with regions of disorder found in the calculated NMR ensemble. The results obtained provide a novel basis to interpret the thioredoxin dependence of the activation of chloroplast NADP-malate dehydrogenase. The specific catalytic mechanism that takes place in the active site of thioredoxins is also discussed on the basis of the recent new understanding and especially in the light of the dual general acid-base catalysis exerted on the two cysteines of the redox active site. It is proposed that the two cysteines of the redox active site may insulate each other from solvent attack by specific packing of invariable hydrophobic amino acids. PMID- 11025546 TI - Assignment of the contribution of the tryptophan residues to the spectroscopic and functional properties of the ribotoxin alpha-sarcin. AB - alpha-Sarcin, a potent cytotoxic protein from Aspergillus giganteus, contains two tryptophan residues at positions 4 and 51. Two single, W4F and W51F, and the double mutant, W4/51F, have been produced and purified to homogeneity. These two residues are neither required for the highly specific ribonucleolytic activity of the protein on the ribosomes (production of the so called alpha-fragment) nor for its interaction with lipid membranes (aggregation and fusion of vesicles), although the mutant forms involving Trp-51 show a decreased ribonuclease activity. Proton NMR data reveal that no significant changes in the global structure of the enzyme occur upon replacement of Trp-51 by Phe. Substitution of each Trp residue results in a 4 degrees C drop in the thermal denaturation midpoint, and the double mutant's midpoint is 9 degrees C lower. Trp-51 is responsible for most of the near-UV circular dichroism of the protein and also contributes to the overall ellipticity of the protein in the peptide bond region. Trp-51 does not show fluorescence emission. The membrane-bound proteins undergo a thermal denaturation at a lower temperature than the corresponding free forms. The interaction of the protein with phospholipid bilayers promotes a large increase of the quantum yield of Trp-51 and its fluorescence emission is quenched by anthracene incorporated into the hydrophobic region of such bilayers. This indicates that the region around this residue is located in the hydrophobic core of the bilayer following protein-vesicle interaction. PMID- 11025547 TI - X-ray crystallographic study of xylopentaose binding to Pseudomonas fluorescens xylanase A. AB - The structure of the complex between a catalytically compromised family 10 xylanase and a xylopentaose substrate has been determined by X-ray crystallography and refined to 3.2 A resolution. The substrate binds at the C terminal end of the eightfold betaalpha-barrel of Pseudomonas fluorescens subsp. cellulosa xylanase A and occupies substrate binding subsites -1 to +4. Crystal contacts are shown to prevent the expected mode of binding from subsite -2 to +3, because of steric hindrance to subsite -2. The loss of accessible surface at individual subsites on binding of xylopentaose parallels well previously reported experimental measurements of individual subsites binding energies, decreasing going from subsite +2 to +4. Nine conserved residues contribute to subsite -1, including three tryptophan residues forming an aromatic cage around the xylosyl residue at this subsite. One of these, Trp 313, is the single residue contributing most lost accessible surface to subsite -1, and goes from a highly mobile to a well-defined conformation on binding of the substrate. A comparison of xylanase A with C. fimi CEX around the +1 subsite suggests that a flatter and less polar surface is responsible for the better catalytic properties of CEX on aryl substrates. The view of catalysis that emerges from combining this with previously published work is the following: (1) xylan is recognized and bound by the xylanase as a left-handed threefold helix; (2) the xylosyl residue at subsite -1 is distorted and pulled down toward the catalytic residues, and the glycosidic bond is strained and broken to form the enzyme-substrate covalent intermediate; (3) the intermediate is attacked by an activated water molecule, following the classic retaining glycosyl hydrolase mechanism. PMID- 11025548 TI - A comparative structural analysis of the ADF/cofilin family. AB - Actin-depolymerizing factor (ADF) and cofilin define a family of actin-binding proteins essential for the rapid turnover of filamentous actin in vivo. Here we present the 2.0 A crystal structure of Arabidopsis thaliana ADF1 (AtADF1), the first plant crystal structure from the ADF/cofilin (AC) family. Superposition of the four AC isoform structures permits an accurate sequence alignment that differs from previously reported data for the location of vertebrate-specific inserts and reveals a contiguous, vertebrate-specific surface opposite the putative actin-binding surface. Extending the structure-based sequence alignment to include 30 additional isoforms indicates three major groups: vertebrates, plants, and "other eukaryotes." Within these groups, several structurally conserved residues that are not conserved throughout the entire AC family have been identified. Residues that are highly conserved among all isoforms tend to cluster around the tryptophan at position 90 and a structurally conserved kink in alpha-helix 3. Analysis of surface character shows the presence of a hydrophobic patch and a highly conserved acidic cluster, both of which include several residues previously implicated in actin binding. PMID- 11025549 TI - Exhaustive mutagenesis in silico: multicoordinate free energy calculations on proteins and peptides. AB - We have extended and applied a multicoordinate free energy method, chemical Monte Carlo/Molecular Dynamics (CMC/MD), to calculate the relative free energies of different amino acid side-chains. CMC/MD allows the calculation of the relative free energies for many chemical species from a single free energy calculation. We have previously shown its utility in host:guest chemistry (Pitera and Kollman, J Am Chem Soc 1998;120:7557-7567)1 and ligand design (Eriksson et al., J Med Chem 1999;42:868-881)2, and here demonstrate its utility in calculations of amino acid properties and protein stability. We first study the relative solvation free energies of N-methylated and acetylated alanine, valine, and serine amino acids. With careful inclusion of rotameric states, internal energies, and both the solution and vacuum states of the calculation, we calculate relative solvation free energies in good agreement with thermodynamic integration (TI) calculations. Interestingly, we find that a significant amount of the unfavorable solvation of valine seen in prior work (Sun et al., J Am Chem Soc 1992;114:6798-6801)3 is caused by restraining the backbone in an extended conformation. In contrast, the solvation free energy of serine is calculated to be less favorable than expected from experiment, due to the formation of a favorable intramolecular hydrogen bond in the vacuum state. These monomer calculations emphasize the need to accurately consider all significant conformations of flexible molecules in free energy calculations. This development of the CMC/MD method paves the way for computations of protein stability analogous to the biochemical technique of "exhaustive mutagenesis." We have carried out just such a calculation at position 133 of T4 lysozyme, where we use CMC/MD to calculate the relative stability of eight different side-chain mutants in a single free energy calculation. Our T4 calculations show good agreement with the prior free energy calculations of Veenstra et al. (Prot Eng 1997;10:789-807)4 and excellent agreement with the experiments of Mendel et al. (Science 1992;256:1798-1802). PMID- 11025550 TI - High performance liquid chromatography-electrospray mass spectrometry for the simultaneous resolution and identification of intrinsic thylakoid membrane proteins. AB - In higher plants, both photosystem I (PSI) and II (PSII) consist of membrane embedded proteins that contain more than one transmembrane alpha helix. PSI is a multiprotein complex consisting of a core complex of thirteen proteins surrounded by four different types of light harvesting antenna proteins. Up to now, the protein components of both photosystems have been characterized by SDS-PAGE and/or immunoblotting and, therefore, identification made only on the basis of electrophoretic mobility, which is sometimes not sufficient to discriminate between individual membrane proteins. This is also complicated by the fact that some proteins, such as the antenna proteins, have almost identical molecular mass and amino acid sequence, making it difficult to identify and ascertain the relative stoichiometry of the proteins. In this paper, we report the complete resolution of the antenna proteins and most of the core components of PSI from spinach, together with the identification of proteins by molecular mass, successfully deduced by the combined use of HPLC coupled on-line with a mass spectrometer equipped with an electrospray ion source (ESI-MS). The proposed RP HPLC-ESI-MS method holds several advantages over SDS-PAGE, including better protein separation, especially for antenna proteins, mass accuracy, speed, efficiency, and the potential to reveal isomeric forms. Moreover, the molecular masses determined by HPLC-ESI-MS are in good agreement with the molecular masses of the individual components calculated on the basis of their nucleotide-derived amino acid sequences, indicating an absence of post-translational modifications in these proteins. It follows that if the method proposed is useful for these highly hydrophobic proteins, it may be of general use for any membrane proteins, where the presence of detergent for solubilization may compromise their characterization. PMID- 11025552 TI - Why are "natively unfolded" proteins unstructured under physiologic conditions? AB - "Natively unfolded" proteins occupy a unique niche within the protein kingdom in that they lack ordered structure under conditions of neutral pH in vitro. Analysis of amino acid sequences, based on the normalized net charge and mean hydrophobicity, has been applied to two sets of proteins: small globular folded proteins and "natively unfolded" ones. The results show that "natively unfolded" proteins are specifically localized within a unique region of charge hydrophobicity phase space and indicate that a combination of low overall hydrophobicity and large net charge represent a unique structural feature of "natively unfolded" proteins. PMID- 11025553 TI - In this issue PMID- 11025551 TI - Aldose and aldehyde reductases: correlation of molecular modeling and mass spectrometric studies on the binding of inhibitors to the active site. AB - Aldose and aldehyde reductases are monomeric NADPH-dependent oxidoreductases that catalyze the reduction of a wide variety of aldehydes and ketones to their corresponding alcohols. The overall three-dimensional structures of the enzymes are composed of similar alpha/beta TIM-barrels, and the active site residues Tyr 50, His 113, and Trp 114 interacting with the hydrophilic heads of inhibitors are conserved. We have used molecular modeling and mass spectrometry to characterize the interactions between the enzymes and three aldose reductase inhibitors: tolrestat, sorbinil, and zopolrestat. Unlike the IC(50) values (concentration of inhibitor giving 50% of inhibition in solution), the Vc(50) values measured by mass spectrometry (accelerating voltage of ions needed to dissociate 50% of a noncovalent complex in the gas phase) for the two enzymes are similar, and they correlate with the electrostatic and hydrogen-bonding energies calculated between the conserved Tyr 50, His 113, and Trp 114 and the inhibitors. The results of our comparison agree with detailed structural information obtained by X-ray crystallography, suggesting that nonconserved residues from the C-terminal loop account for differences in IC(50) values for the two enzymes. Additionally, they confirm our previous assumption that the Vc(50) values reflect the enzyme inhibitor electrostatic and hydrogen-bonding interactions and exclude the hydrophobic interactions. PMID- 11025554 TI - Oral insulin and the induction of tolerance in man: reality or fantasy? AB - Induction of tolerance to insulin, the only beta-cell-specific antigen in Type 1 diabetes, is under testing for prevention of Type 1 diabetes in the US multicentre trial DPT1. Recently a multicentre double-blind trial with oral insulin in patients with recent onset Type 1 diabetes, conducted by our group, has been completed and showed that oral insulin administration at the dose of 5 mg daily for one year starting at the time of disease onset had no effect on residual beta-cell function as assessed by C-peptide secretion. A similar trial using different doses was carried out at the same time and similarly showed no beneficial effect on the decline of beta-cell function during the first year after diagnosis. In this study oral insulin was administered at the daily doses of 2.5 and 7. 5 mg over a one-year period. Such results challenge the current view that induction of oral tolerance can be established when the immune process is already active. PMID- 11025555 TI - Why is learning and memory dysfunction in Type 2 diabetes limited to older adults? AB - Review of the literature on the cognitive correlates and consequences of Type 2 diabetes reveals two very intriguing findings. Not only are verbal learning and memory skills most likely to be disrupted as compared to other cognitive skills (e.g. attention, executive function; psychomotor efficiency), but these mnestic deficits appear to be restricted to individuals with diabetes who are older than 60-65 years of age. Middle-aged adults with either Type 2 or Type 1 diabetes are apparently protected insofar as researchers have only infrequently reported learning and memory impairments in that age group. Why do older adults have such an increased risk of diabetes-associated memory dysfunction? In our view, this phenomenon is a consequence of a synergistic interaction between diabetes-related metabolic derangements and the structural and functional changes occurring in the central nervous system (CNS) that are part of the normal ageing process. To critically evaluate that possibility, we summarise what is known about learning and memory dysfunction in the adult with diabetes, examine the extent to which chronic hyperglycaemia may adversely affect the integrity of the CNS, and selectively review the literature on age-associated changes in brain morphology and cognitive function, paying special attention to the threshold theory of cognitive impairment. PMID- 11025556 TI - Cerebral oedema during treatment of diabetic ketoacidosis: are we any nearer finding a cause? AB - Cerebral oedema remains the leading cause of death and morbidity in children with Type 1 diabetes mellitus. Around seven per thousand episodes of diabetic ketoacidosis (DKA) are complicated by cerebral oedema, and one-quarter of those children will die from it. The cause or causes of cerebral oedema are still very poorly understood, but lawyers are already keen to implicate various aspects of the management of DKA. There have been many theories as to the pathophysiology of cerebral oedema, and possible contributing factors may be excessive rate of rehydration, falling plasma osmolality (particularly that due to a reduction in plasma sodium concentration), hypoxia and insulin dosage. There is some supportive evidence for all of these factors in some cases, but there have been no sizeable case-control studies, in part because of the rarity of the condition. Furthermore, cerebral oedema can still occur even when the management of DKA follows current 'best practice' guidelines. As the mechanisms of cell volume regulation within the brain are increasingly understood, different questions may provide greater insights. For example, what is it about children that makes them so much more susceptible to cerebral oedema than adults? And why does one child treated in a certain way develop cerebral oedema whereas another does not? The anxiety over causing cerebral oedema has driven most of the changes in the management of DKA over recent decades, yet there is no evidence that the incidence has reduced. Until the causes are understood, we cannot be dogmatic about treatment recommendations. PMID- 11025557 TI - The role of the proteasome in autoimmunity. AB - Type 1 diabetes is believed to be caused by T cell-mediated autoimmunity, with a prediabetic state characterized by the production of autoantibodies specific for proteins expressed by pancreatic beta cells. The non-obese diabetic (NOD) mouse is a spontaneous model of Type 1 diabetes with a strong genetic component that maps to the major histocompatibility complex (MHC) region of the genome. A specific proteasome defect has now been identified in NOD mouse lymphocytes that results from down-regulation of expression of the proteasome subunit LMP2, which is encoded by a gene in the MHC genomic region. This defect both prevents the proteolytic processing required for the production and activation of the transcription factor nuclear factor-kappaB (NF-kappaB), which plays an important role in immune and inflammatory responses, in addition to increasing the susceptibility of the affected cells to apoptosis induced by tumor necrosis factor-alpha (TNF-alpha). The proteasome dysfunction is both tissue- and developmental stage-specific and likely contributes to disease pathogenesis and tissue targeting. PMID- 11025558 TI - The molecular specificity of insulin autoantibodies. AB - Insulin autoantibodies (IAA) are one of several markers for Type I (autoimmune) diabetes, but alone deserve special attention. Unlike the other markers, their ligand is unique to the beta cell. IAA are the first markers to appear during the symptomless period which precedes diabetes and they are present in the vast majority of young children destined to develop diabetes. The primary and tertiary structures of insulin have been known for decades. Binding studies with insulin variants have shown epitope restriction that can distinguish Type 1 diabetes predictive from non-predictive IAA-positive sera, thereby improving specificity for the test. With two major international Type 1 diabetes prevention trials underway, there is a pressing need to refine markers that reliably indicate the presence of, and remission from, autoimmune insulitis. The binding regions of antibodies are assembled from three multi-gene families, and some of their diversity derives from random mutation during their antigen-driven maturation. There is evidence that mature IAA derive from germline-encoded 'natural' antibodies, and that the gene segments utilised by IAA may be influenced by clinical context. Monoclonal anti-idiotypic (anti-Id) antibodies can serve as probes for antibody variable region determinants, and antibodies to the different epitopes of beef and porcine insulins have already been analysed with monoclonal reagents. Used as antibodies in a radioimmunoassay format, monoclonal anti-Ids will identify and measure autoantibody idiotopes as if they were ligands. The challenge now is to replace the conventional radiobinding assays for IAA, which only detect and titrate, with radioimmunoassays that can be standardised in absolute units. There is sufficient evidence for the existence of Type 1 diabetes predictive IAA idiotopes to justify the development of idiotope-specific radioimmunoassays which ignore Type 1 diabetes-unrelated IAA. PMID- 11025560 TI - Subcutaneous adipose tissue expression of plasminogen activator inhibitor-1 (PAI 1) in nondiabetic and Type 2 diabetic subjects. AB - OBJECTIVE: Increased plasma levels of plasminogen activator inhibitor-1 (PAI-1) have been suggested to be a part of the insulin resistance syndrome, and recent data suggest that adipose tissue participates in the production of PAI-1. We examined the expression and insulin regulation of subcutaneous adipose tissue PAI 1 mRNA and its relationship to insulin sensitivity. DESIGN: A cross-sectional study involving five lean (60.0+/-3.1 years, BMI 23.5+/-0.5 kg/m(2)) and six obese nondiabetic men (56.0+/-3.1 years, BMI 30.4+/-0.7 kg/m(2)), and six obese Type 2 diabetic men (61.4+/-3.2 years, BMI 31.8+/-1.0 kg/m(2)). MEASUREMENTS: Subcutaneous adipose tissue PAI-1 mRNA and insulin sensitivity were quantified using RT-competitive PCR and euglycemic hyperinsulinemic clamp technique, respectively. RESULTS: Subcutaneous adipose tissue PAI-1 mRNA levels were higher in obese nondiabetic and Type 2 diabetic men than in lean nondiabetic men. PAI-1 mRNA levels decreased in the three groups during a 240-min euglycemic hyperinsulinemic clamp (P<0.05 for all groups), and a similar reduction was observed during a 240-min saline control study indicating that adipose tissue PAI 1 gene expression has diurnal variation and is not acutely controlled by hyperinsulinemia. The basal PAI-1 mRNA levels correlated positively with BMI, and waist-to-hip ratio; and negatively with whole-body glucose disposal rate in nondiabetic men. CONCLUSIONS: Subcutaneous adipose tissue PAI-1 mRNA expression is increased in obese nondiabetic or in Type 2 diabetic men. Subcutaneous adipose tissue PAI-1 mRNA expression is increased in proportion to visceral obesity and to the level of whole-body insulin resistance. Subcutaneous adipose tissue PAI-1 mRNA expression is not acutely regulated by insulin, and it is subject to a diurnal variation. PMID- 11025559 TI - Insulin receptor in rat peripheral nerve: its localization and alternatively spliced isoforms. AB - BACKGROUND: Diabetic neuropathy accompanies both Type 1 and Type 2 diabetes, although it shows in both humans and animal models distinct differences between the two types of diabetes. Progressive paranodal degenerations occurring in Type 1, but not in Type 2, diabetes is believed to account for the more severe functional deficits in Type 1 diabetic rats. This suggests that factors other than hyperglycemia, such as insulin deficiency, may play a pathogenetic role. In this study, we investigated the immunolocalization of the insulin receptor (IR) and the expression of its two alternatively spliced isoforms in adult rat peripheral nerve. METHODS: Adult male Wistar rats 6-8 months of age were examined. Both light and ultrastructural immunohistochemistry was employed for localization of IR. The antibody was a mouse monoclonal antibody raised against the beta-subunit of human IR. Reverse transcription polymerase chain reaction (RT PCR) was used to identify the two IR isoforms in peripheral nerve and seven other organs. Localization of the mRNA message was assessed by in situ hybridization. RESULTS: IR was localized to paranodal terminal Schwann cell loops and microvilli and to the paranodal axolemma. Furthermore, IR immunoreactivity was also present in Schmidt-Lantermann incisures. Endoneurial vessels showed IR localization on plasma membranes and in endocytotic vesicles of endothelial cells and pericytes. A high intensity of immunostained IR was found in close proximity to interendothelial tight junctions. Peripheral nerve showed, like the brain, predominantly the high affinity IR lacking exon 11. The mRNA message was localized to Schwann cells, endothelial cells and pericytes. CONCLUSION: Peripheral nerve expresses predominantly the high affinity IR, which is localized to strategic structures associated with the blood-nerve barrier and the paranodal ion-channel barrier. PMID- 11025562 TI - Current literature in diabetes. PMID- 11025561 TI - Molecular scanning for mutations in the insulin receptor substrate-1 (IRS-1) gene in Mexican Americans with Type 2 diabetes mellitus. AB - BACKGROUND: Insulin receptor substrate-1 (IRS-1) is an endogenous substrate for the insulin receptor tyrosine kinase, which plays an important role in insulin signaling. Mutations in the IRS-1 gene are associated in some populations with obesity and Type 2 diabetes. METHODS: To determine whether variation in the IRS-1 gene contributes to genetic susceptibility to insulin resistance and Type 2 diabetes in Mexican Americans, the entire coding region of the IRS-1 gene was screened for variation in 31 unrelated subjects with Type 2 diabetes using single stranded conformational polymorphism analysis (SSCP) and dideoxy sequence analysis. Variants encoding amino acid substitutions were genotyped in 27 unrelated nondiabetic Mexican Americans and in all family members of subjects containing these variants, and association analyses were performed. To trace the ancestral origins of the variants, Iberian Caucasians and Pima Indians were also genotyped. RESULTS: Eight single base changes were found: four silent polymorphisms and four missense mutations (Ala94Thr, Ala512Pro, Ser892Gly and Gly971Arg). Allele frequencies were 0.009, 0.017, 0.017 and 0.043, respectively. There were no significant associations of any of these variants with diabetes, glucose or insulin levels during an oral glucose tolerance test, or with body mass index (BMI) in Mexican American families except for a modest association between the Ala94Thr variant and decreased BMI (30.4 kg/m(2) vs 24.0 kg/m(2); p=0.035). None of these four missense mutations were detected in Pima Indians. In Iberian Caucasians, neither Ala94Thr nor Ser892Gly were detected, and Ala512Pro was detected in only 0/60 diabetic patients and 1/60 nondiabetic controls. Gly971Arg was relatively more common in Iberian Caucasians with 12/58 diabetic patients and 7/60 nondiabetic controls being heterozygous for this variant (p=0.21 for comparison between diabetic and nondiabetic subjects). CONCLUSIONS: Ala94Thr, Ala512Pro and Ser892Gly mutation are rare in the populations studied. Gly971Arg, is more common in Mexican Americans and Caucasians, but is not a major contributor to genetic susceptibility to Type 2 diabetes. PMID- 11025563 TI - Prognostic value of recurrent episodes of creatine kinase-MB elevation following repeated catheter-based coronary interventions. AB - Creatine kinase-MB (CK-MB) enzyme elevations were shown to affect cardiac prognosis following percutaneous coronary interventions (PCIs). This study examined whether recurrent episodes of CK-MB elevation following repeated PCIs may be associated with a cumulative adverse prognostic risk. We studied 767 consecutive patients (age, 64 +/- 11 years; 69% male) who underwent two consecutive PCI procedures on two separate hospitalizations (mean interval, 121 +/- 110 days). Patients were stratified into four groups according to number of episodes of any (> 4 ng/ml) postinterventional CK-MB rise (no elevation, previously elevated, currently elevated, or elevated at the time of both procedures; n = 403, 107, 153, and 104 patients, respectively). In-hospital clinical outcomes (death, Q-MI, and repeat revascularization) and up to 1-year follow-up events were obtained. Recurrent episodes of CK-MB elevation were associated with increased in-hospital mortality (3.8% vs. 0.9% vs. 0% vs. 0%, P = 0.0003), increased cumulative mortality (18.9% vs. 5.9% vs. 4.3% vs. 4.3%, P = 0.0003) and cumulative Q wave MI (8.0% vs. 4.9% vs. 1.0% vs. 0.8%, P = 0.005) at 1 year, and lower overall cardiac event-free survival at follow-up (66.8% vs. 80.5% vs. 88.8% vs. 88.8%, P = 0.0001 for patients with twice, current, previous, and no CK-MB elevation, respectively). By multivariate analysis, CK-MB elevated at the time of both procedures, was the strongest independent predictor for cumulative mortality (OR 3.4, 95% CI 1.6-7.1, P = 0.001) or any adverse cardiac events (OR 2.6, 95% CI 1.6-4.3, P = 0.0002). We conclude that cumulative episodes of periprocedural CK-MB elevation are associated with an incremental adverse prognostic risk including mortality and Q-wave MI. Thus, measures aimed at reducing subsequent CK-MB rise may be warranted in particular among patients with a prior history of PCI related CK-MB elevation. PMID- 11025564 TI - Evaluation of the role of abciximab (Reopro) as a rescue agent during percutaneous coronary interventions: in-hospital and six-month outcomes. AB - Abciximab is effective for the prevention of complications when administered prior to percutaneous coronary intervention (PCI). The efficacy and safety of abciximab as an unplanned or rescue agent for complications of PCI is unknown. Rescue versus planned use was compared in 186 consecutive patients. Primary or rescue PCI for acute myocardial infarction (MI) and shock were excluded. Rescue abciximab use was undertaken in 101 patients (54.3%) and planned abciximab was used in 85 (45.7%). The rescue abciximab patients had a lower incidence of previous MI, preprocedural thrombus, multivessel, and vein graft intervention. In hospital endpoints in the rescue versus planned abciximab patients were death (1.0% vs. 1. 2%, P = 1.0), Q-wave MI (2.0% vs. 2.4%, P = 1.0), any MI (14.9% vs. 9.4%, P = 0.3), target vessel revascularization (TVR; 0% vs. 1.2%, P = 1.0), and composite (15.8% vs. 10.6%, P = 0.3). At 6 months, events were death (4.0% vs. 2.3%, P = 0.69), MI (14.9% vs. 9.4%, P = 0.26), TVR (20.8% vs. 4.7%, P = 0.001), and composite (30.7% vs. 15. 3%, P = 0.01). In-hospital complications between the rescue and planned abciximab patients of major bleed (1.0% vs. 1.8%, P = NS), stroke (0% vs. 1.8%, P = NS), and thrombocytopenia (3.0% vs. 1.8%, P = NS) were similar. There was a significantly higher procedural time (99.6 min vs. 86.1 min, P = 0.02), contrast volume (278.8 ml vs. 223. 5 ml, P = 0.04), and heparin use (8984 u vs. 6003 u, P = 0.0006) in the rescue group. In this nonrandomized comparison, rescue abciximab allowed for the safe discharge from hospital in the majority of patients. However, during a 6-month follow-up, more patients treated with rescue abciximab required TVR with either repeat PCI or CABG. Further studies are warranted to evaluate the overall strategy of rescue abciximab use in PCI. PMID- 11025565 TI - "IIb or not IIb or maybe IIb?" Is that the question? Or is it "which IIb and to whom?". PMID- 11025566 TI - How long is enough? Defining the treatment length in endovascular brachytherapy. AB - The International Commission on Radiation Units and Measurement (IRCU) 50 has clearly defined treatment volumes in radiation therapy in the management of neoplasms. These concepts are applied to the field of endovascular brachytherapy (EVBT) for the prevention of postangioplasty restenosis. The following definitions are proposed: gross target length (GTL) is defined as the narrowed segment of the artery that requires intervention. Clinical target length (CTL) is defined as the intervened or injured length, which could be due to angioplasty, stent strut injury, stent deployment, or debulking procedures. Planning target length (PTL) is the CTL plus a margin to account for heart/catheter movement and uncertainty in target localization. The final treatment length (TL) is the PTL plus the effect of penumbra. The accurate specification of treatment length serves several important purposes. Based on an understanding of the different factors constituting the treatment length, adequate margins can be provided beyond the GTL; this will avoid geographic misses and minimize edge failures. These definitions of target length ensure treatment consistency and provide a standard terminology for communication among practitioners of EVBT, something of critical importance in the conduct of multi-institutional trials in this new and multidisciplinary therapy. Finally, since the efficacy of EVBT is critically dependent on the precision of radiation delivery, these guidelines ensure that the benefits of EVBT seen in prospective randomized trials can be translated into daily clinical practice at the community level. PMID- 11025567 TI - Intracoronary brachytherapy not associated with changes in major side branches. AB - Intravascular radiation (IR) is emerging as a potential solution for the prevention and treatment of restenosis. Subacute thrombosis late coronary occlusion of the target lesion after intracoronary brachytherapy has been described in two studies. None of these studies assessed in detail the evolution of side branches following IR. The purpose of this study was to determine the frequency of side-branch occlusion in patients undergoing IR. We evaluated the procedural and 6-month follow-up angiograms of 170 consecutive patients enrolled in the GAMMA-I and WRIST studies who presented a major side branch (diameter > 1.5 mm) at the treated lesion. Major side branches treated during the procedure and/or total occlusion of the main branch were excluded for analysis. Sixty-three patients (control = 30, irradiated = 33) were identified and side branches were classified in one of three categories: no significant stenosis, > 50% stenosis, and occluded. The baseline patient and lesion characteristics were similar among the classified groups. No difference was observed between the control and irradiated arteries after the procedure for side-branch occlusion (14% and 9%, P = NS). A significant decrease of restenosis was observed after IR compared to control (20% vs. 70%: P < 0.001). The patency rates and rates of side-branch occlusion were similar between the two groups. However, in the control group, 50% of the side-branch occlusions that occurred after the procedure were patent at 6 month follow-up. In contrast, in the irradiated group, the incidence of side branch occlusion increased from 9% to 15% at 6-month follow-up in the irradiated group. Low doses of radiation applied to noninjured side branches are not associated with adverse effects. The absence of restoration patency followed by IR is most likely a consequence of a delayed healing process. PMID- 11025568 TI - Cardiac catheterization in diagnosis and management of congenital heart disease in thoracopagus conjoined twins. AB - Over the last two decades, surgical separation of thoracopagus conjoined twins has become increasingly feasible. Delineation of the degree of cardiac involvement is essential to establish the feasibility of surgical separation. We describe our experience of cardiac catheterization both in the diagnosis and management of nine sets of thoracopagus twins who presented to a single center over three decades and discuss its role in conjunction with other modern diagnostic modalities. PMID- 11025569 TI - Congenital severe aortic stenosis with congestive heart failure in late childhood and adolescence: effect on left ventricular function after balloon valvuloplasty. AB - Left ventricular (LV) dysfunction with congestive heart failure (CHF) resulting from severe congenital aortic stenosis (AS) is a well-described condition in infancy, but it is rarely found in older children and adolescents. Aortic valve surgery in such cases may be associated with higher rates of morbidity and mortality. Aortic valve balloon dilatation (AVBD) is a viable alternative, but its effect on LV function has not been evaluated. We describe follow-up results of AVBD in 10 cases of severe congenital AS in older children and adolescents with CHF and LV dysfunction. The ages of these patients ranged from 5 to 18 yr (mean +/- SD: 10.8 +/- 4 yr), and nine were males. The follow-up period after AVBD ranged from 3 mo to 7 yr (mean +/- SD: 2.93 +/- 2.1 yr). Success was achieved in all cases, with no immediate complications. After valvuloplasty, the peak-to-peak systolic gradient declined from 74.7 +/- 30.8 to 33.9 +/- 18.2 mm Hg (P < 0.0001). The cardiac index increased slightly but significantly, from 1.9 +/ 0.27 to 2.2 +/- 0.5 L/min/m(2) (P < 0.015). Hemodynamic improvement was also confirmed by a significant decrease in mean pulmonary artery and pulmonary artery wedge pressures from 41.9 +/- 9 to 32.6 +/- 6.6 and from 25.5 +/- 2.9 to 19.3 +/- 3.4 mm Hg, respectively. The echocardiographically derived left ventricular ejection fraction (LVEF) improved from 21.6 +/- 5. 37% to 31 +/- 6.5% within 24 hr after AVBD, and it further improved in all cases on follow-up. Mean LVEF at last follow-up was 59.4 +/- 11.4%. The Doppler instantaneous peak systolic gradient (IPSG) increased from 37.3 +/- 18.8 to 64.8 +/- 30.7 mm Hg at late follow-up. Significant aortic regurgitation (AR) developed in 20% of patients. The Doppler IPSG across the aortic valve was > 60 mm Hg in five cases on follow up. Two of these patients underwent another AVBD successfully 4 and 16 mo later, respectively. Aortic valve replacement was done in two patients, one for severe restenosis with mild AR 12 mo after AVBD and another for severe re-restenosis with moderate AR 21 mo after a second AVBD. Severe congenital AS can be associated with LV dysfunction and CHF in late childhood and adolescence. AVBD results in good palliation with improvement in LV function on follow-up. PMID- 11025570 TI - Balloon aortic valvuloplasty: triumphs again. PMID- 11025571 TI - Modified Judkins catheters for selective coronary angiography in infants and young children. AB - The purpose of this study was to assess 4 Fr Judkins catheters with modified shorter tips for performing selective coronary angiography in infants and young children. Twenty patients ranging 6 weeks to 3. 8 years of age were enrolled. Retrograde left heart catheterization and selective coronary angiography were performed. Right (JR) and left (JL) catheters with modified 1.5- and 2.5-cm curves (Cordis) were used. Thirty-six of 37 coronary arteries were successfully cannulated and demonstrated. Median procedure time was 95 sec for the right and 50 sec for the left coronary artery. Median fluoroscopy time was 1.1 min for the right and 0.7 min for the left coronary artery. The JL 1.5 appeared best suited for patients less than 75 cm tall. The JR 1.5 was suitable for patients up to 85 cm tall. Taller patients required the 2.5-cm curves. It is concluded that these modified 4 Fr Judkins catheters were effective. PMID- 11025572 TI - Transcatheter reconstruction of obliterated superior vena cava with additional obliteration of right internal jugular and subclavian vein. AB - We describe the successful reconstruction of a completely obliterated superior vena cava, despite additional obliteration of right internal jugular and subclavian vein, using an end-to-side approach to connect to the innominate and left internal jugular vein. We also document long-term patency. PMID- 11025573 TI - Postsurgical use of amplatzer septal occluder in cyanotic patients with pulmonary atresia/intact ventricular septum: significance of cor triatriatum dexter and dilated right atrium. AB - Percutaneous closure of secundum atrial septal defects (ASDs) has been shown to be safe and effective. However, its role after surgery in patients with cyanotic congenital heart disease who may have associated cor triatriatum dexter and a dilated right atrium has not been established. This article reports on successful closure in such patients, including precautions and results. PMID- 11025574 TI - New interventional therapeutic approach for dual drainage of the scimitar vein. AB - A patient with scimitar syndrome and dual drainage of a right-sided scimitar vein into the inferior vena cava and the left atrium underwent coil occlusion of the right aortopulmonary collateral artery and device occlusion of the lower scimitar vein drainage, leaving it flowing solely into the left atrium. PMID- 11025575 TI - Successful conservative treatment of an infected central venous stent. AB - Several cases of stent infection have been reported in the medical literature. Most required surgical removal of the stent, sometimes with severe sequelae including limb loss. We report the case of a 39-year-old hemodialysis patient who, 3 weeks prior to admission, had undergone angioplasty of the right innominate and subclavian veins with implantation of a Wallstent. He was admitted to the hospital with sepsis due to Staphylococcus aureus and had histological evidence of endovascular infection within the stent. The patient was successfully treated with a 6-week course of intravenous antibiotics delivered directly into the stent, thus avoiding the need for surgical removal of the device. PMID- 11025576 TI - Intracoronary irradiation and stent placement in a chronic total coronary occlusion: long-term clinical, angiographic, and intracoronary ultrasound follow up. AB - Intracoronary irradiation emerges as a promising method in a variety of restenosis prone coronary lesions. We report the acute and long-term clinical, angiographic, and ICUS follow-up of a patient who underwent a successful angioplasty with stent placement in a chronic coronary occlusion with adjuvant gamma-intracoronary radiation. PMID- 11025577 TI - Prophylactic brachytherapy. PMID- 11025578 TI - Left main coronary artery aneurysm: sealing by stent graft and long-term follow up. AB - We describe the implantation of a stent graft to seal off the left main coronary artery aneurysm of a 64-year-old man with guidance by intravascular ultrasound. The aneurysm was successfully sealed off. At 6-month and 1-year follow-up, a small residual aneurysm reappeared, which was treated conservatively. The technical issues in closing this aneurysm and the management of the residual aneurysm are discussed. PMID- 11025579 TI - Nonsurgical closure of a patent foramen ovale in a patient with carcinoid heart disease and severe hypoxia from interatrial shunting. AB - We report the percutaneous transcatheter closure of a patent foramen ovale using an Amplatzer septal occluder in a rare patient with carcinoid heart disease involving both the right and left heart who presented with severe hypoxemia secondary to intra-atrial shunting. We believe this is the first report of this technique being utilized in a patient with carcinoid heart disease and it may represent an alternative to surgical closure in these patients at high risk for surgical complications. PMID- 11025580 TI - An unusual mechanism of coronary stent dislodgment and localization. AB - Stent dislodgment from its delivery catheter is a rare yet clinically important entity in cardiac catheterization. We report a highly unusual case of stent loss and subsequent localization in the guiding catheter. Our hypothesized mechanism of stent dislodgment should be recognized by the medical community. PMID- 11025581 TI - Compartment syndrome from balloon pump. AB - A 35-year-old man had significant left main coronary artery disease and required intra-aortic balloon catheter insertion owing to refractory ischemia before emergency coronary artery bypass graft. In the immediate postoperative period the patient started complaining of leg pain and diminished sensation in the right foot despite palpable pulses. The diagnosis of acute compartment syndrome of the right leg was made by an intracompartment pressure measurement of 90 mm Hg. PMID- 11025582 TI - Transcatheter occlusion of large pulmonary arteriovenous fistula. AB - We describe the compound transcatheter occlusion of a large and symptomatic pulmonary arteriovenous malformation in a 58-yr-old man. Pre- and postintervention clinical and laboratory data support the impression of an excellent outcome. PMID- 11025583 TI - Stent placement for non-STRESS/BENESTENT lesions: a critical review. AB - Stenting lesions with favorable characteristics as required for inclusion in the STRESS/BENESTENT trials have yielded superior results to that of PTCA alone. Results for less favorable lesions such as in small vessels, diffuse disease, ostial disease, and saphenous vein grafts are less well established. This review seeks to analyze available data for stent placement in this subset of non STRESS/BENESTENT lesions. PMID- 11025584 TI - Coronary rotational atherectomy via transradial approach: a study using radial artery intravascular ultrasound. AB - The use of coronary rotational atherectomy via radial artery (RA) has been limited because of the large diameter of guiding catheters. We studied the feasibility of this approach by sizing the RA by intravascular ultrasound (IVUS) and using 7 Fr (2.31 mm) guiding catheters. Seventeen transradial percutaneous transluminal coronary rotational atherectomy (PTCRA) procedures were performed in 16 patients, mean age 62 +/- 12 years, for a total of 19 vessels treated. The mean RA diameter was 2.9 +/- 0.36 mm and the mean reference diameter of the treated coronary vessels was 2.7 +/- 0.45 mm. The mean coronary percent stenosis was 74% +/- 10%, the mean minimum lumen diameter was 0.76 +/- 0.35 mm, and the mean lesion length was 16 +/- 19 mm. Ten vessels were treated with rotational atherectomy alone, or with adjunctive high pressure balloon angioplasty, achieving an acute lumen gain of 0.8 +/- 0.4 mm (P = 0. 001). Nine arteries had stent implantation in addition to rotational atherectomy, resulting in an acute lumen gain of 2.4 +/- 0.5 mm (P = 0.001). The success rate was 94%. There were no vascular complications. Two patients had a non-Q myocardial infarction. In conclusion, transradial PTCRA when used in conjunction with IVUS of the RA is a safe and feasible procedure in selected cases. This may be an alternative approach of revascularization technique especially for patients with limited vascular access and for those who require early ambulation or early discharge from the hospital. PMID- 11025585 TI - Coronary angioplasty performed using the FullFlow mechanical dilatation-perfusion catheter: initial animal experience. AB - We report our initial experience with an animal model, using a new mechanical catheter-based dilatation system, the FullFlow (FF) catheter. The primary purpose of this study was to compare coronary flow achieved using the FF perfusion catheter and a traditional balloon angioplasty perfusion catheter. Baseline average peak velocity (APV) measurements were obtained using the FloWire Doppler guide wire. Either a Surpass Perfusion balloon (PB) or an FF catheter was advanced to a coronary segment. Doppler measurements were made before, during, and after full device expansion. Simultaneous heart rate, EKG, and blood pressure readings were recorded. Significant 36.1% and 29% reductions in mean APV were found for PB when comparing baseline to device-up measurements and when comparing device-down to device-up measurements, respectively. In contrast, significant 12.2% and 18.5% increases were seen in mean APV for the FF. No significant differences were found in heart rate, systolic blood pressure, diastolic blood pressure, or ST-segment changes. The FF system produces superior downstream coronary perfusion with side-branch coronary patency compared with results using a contemporary perfusion balloon angioplasty catheter. PMID- 11025586 TI - Loops and transradial approach in coronary diagnosis and intervention. PMID- 11025587 TI - Reply to the letter to the editor by louvard and Lefevre PMID- 11025589 TI - Transplanting bone-marrow stem cells in the central nervous system. PMID- 11025588 TI - Satler L, Mintz G,--patience in the pursuit of perfection. PMID- 11025590 TI - Efficacy of pipobroman in the treatment of polycythemia vera: long-term results in 163 patients. AB - BACKGROUND AND OBJECTIVES: Polycythemia vera (PV) is a myeloproliferative disorder, characterized by the expansion of the red cell mass. Our purpose was to evaluate the efficacy of pipobroman (PB) in the long-term control of PV and to assess early and late events. DESIGN AND METHODS: From June 1975 to December 1997, 163 untreated patients with PV (median age 57 years, range 30-82) were treated with PB in a single Institute for a median follow-up of 120 months. The diagnosis was made according to the Polycythemia Vera Study Group criteria. PB was given at the dose of 1 mg/kg/day until hematologic response (hematocrit < 45% and platelets < 400x109/L) and of 0.3-0.6 mg/kg/day as maintenance therapy. RESULTS: Hematologic remission was achieved in 94% of patients in a median time of 13 weeks (range 6-48). Median overall survival was 215 months, with a standardized mortality ratio of 1.7. The cumulative risk of death was 11%, 22%, and 26% at 7, 10, and 12 years, respectively. The incidence of thrombotic events was 18.4x105 person-year and the cumulative risk was 6%, 11%, 16%, and 20% at 3, 7, 10, and 12 years respectively. Acute leukemia occurred in 11 patients, myelofibrosis in 7, and solid tumors in 11. The 10-year cumulative risk of leukemia, myelofibrosis, and solid tumors was 5%, 4%, and 8%, respectively. In the logistic analysis age over 65 (p = 0.0001) and thrombotic events at diagnosis (p = 0.001) were significantly correlated with a higher risk of death. Female gender (p = 0.02) and age over 65 (p = 0.01) significantly influenced the occurrence of thrombotic complications. Age was the only significant risk factor for leukemia (p = 0.04) and for solid tumors (p = 0.03), while the duration of PB treatment did not influence these risks. No significant risk factor was demonstrated for myelofibrosis. INTERPRETATION AND CONCLUSIONS: This study demonstrates in a large series of patients, observed for a long period, that pipobroman is effective in the long-term control of PV. The risk of early thrombotic complications at 3 years is 6% and the 10-year risk of acute leukemia, late myelofibrosis, and solid tumors is 5%, 4%, and 8%, respectively. The duration of pipobroman treatment did not correlate with these events. PMID- 11025591 TI - Bax mutations are an infrequent event in indolent lymphomas and in mantle cell lymphoma. AB - BACKGROUND AND OBJECTIVES: The Bax gene is one of the most important genes involved in apoptosis regulation. Recently, it has been proposed that inactivating mutations of this death agonist may contribute to the pathogenesis of human tumors. This study was aimed at defining the status of the Bax gene in indolent lymphomas. DESIGN AND METHODS: Fifty paraffin-embedded biopsies from indolent lymphomas (10 small lymphocytic lymphomas, 5 immunocytomas, 20 follicular lymphomas and 15 marginal zone lymphomas) and 10 mantle cell lymphomas ( MCL ) were studied. All six exons of the Bax gene, together with their flanking sequences, underwent mutational analysis by PCR-SSCP followed by direct sequencing of positive cases. Moreover, Bax protein expression was investigated in all samples by immunohistochemical analysis. RESULTS: All analyzed cases showed wild type Bax gene alleles and variable levels of Bax protein expression. INTERPRETATION AND CONCLUSIONS: This study indicates that deregulation of apoptotic control in indolent lymphomas and MCL is not caused by Bax mutations and that other molecular mechanisms must, therefore, be involved. PMID- 11025592 TI - Cytotoxic hepatosplenic gammadelta T-cell lymphoma following acute myeloid leukemia bearing two distinct gamma chains of the T-cell receptor. Biologic and clinical features. AB - BACKGROUND AND OBJECTIVES: Hepatosplenic gd T-cell lymphoma is a rare entity of peripheral T-cell lymphomas. We characterized in detail the first case of hepatosplenic gd -T-cell lymphoma following acute myeloid leukemia. DESIGN AND METHODS: Hepatosplenic gd -T-cell lymphoma was diagnosed in a woman who had been in complete remission (CR) of acute myeloid leukemia (AML) for two years. Improvement but no objective response of the disease was observed after various types of chemotherapy. CR was achieved after related donor stem cell transplantation. Thirteen months later relapse of hepatosplenic gd T-cell lymphoma was diagnosed. While being prepared for a second transplantation the patient developed meningeal lymphoma and died. The patient's lymphoma cells were studied by immunologic, functional and molecular techniques. RESULTS: Lymphoma cells expressed the gd T-cell receptor (TCR), CD2, CD3, CD5, CD7, CD38, CD45, CD161 (NKR-P1), TIA and Ki67. Further analysis revealed expression of Vd1 and two distinct TCRg chains, Vg3 and Vg9, by the malignant cell clone. The clonality of the T-cells was confirmed by reverse transcriptase polymerase chain reaction (RT PCR) followed by sequencing of TCR Vg3, Vg9 and Vd1 junctional regions. Clone specific PCR was negative at diagnosis of AML and was positive at all times during follow-up of the hepatosplenic gd T-cell lymphoma. The lymphoma cells mediated strong natural killer cell-like cytotoxic activity, possibly explained by expression of CD161 and a lack of killer inhibitory receptor. INTERPRETATION AND CONCLUSIONS: Several so far undescribed features were observed in this case of hepatosplenic gd T-cell lymphoma, such as T-cell lymphoma following AML, expression of two distinct T-cell receptor g-chains, and an unexpected cytotoxic phenotype. PMID- 11025594 TI - Factors predicting response to splenectomy in adult patients with idiopathic thrombocytopenic purpura. AB - BACKGROUND AND OBJECTIVES: Splenectomy is the treatment of choice in the majority of patients affected by idiopathic thrombocytopenic purpura refractory to corticosteroid therapy, but it is not free from early and late complications. As the available literature does not seem to contain any precise indications concerning possible factors predicting the response to splenectomy, the aim of this retrospective study of 65 splenectomized patients was to attempt to identify potentially predictive clinical or laboratory parameters. DESIGN AND METHODS: For the purposes of statistical analysis, the patients were divided into two groups: the first included those with a complete (platelets > 100x10(9)/L) or partial response (platelets 50-100 x10(9)/L) to splenectomy; the second, the non responders (platelets < 50x10(9)/L). The non-parametric tests were based on the Kruskal-Wallis method for independent samples, and the independent samples were compared using the Chi-square test according to Pearson. RESULTS: Univariate analysis did not reveal any significant correlation between successful splenectomy and age, sex, platelet count at diagnosis, anti-platelets antibody positivity, the site of platelet sequestration, the time between diagnosis and surgery, or the response to high intravenous immunoglobulin doses. However, the probability of success was greater in the patients with a complete or partial pre operative response to steroid therapy (p<0.05). INTERPRETATION AND CONCLUSIONS: The factor most frequently associated with the success of splenectomy is the site of autologous platelet sequestration. Our study did not identify any clinical or laboratory parameter clearly predictive of post-splenectomy cure other than a transient response to steroid treatment. This finding needs further confirmation in larger patient populations. PMID- 11025593 TI - Treatment of early-stage Hodgkin's disease with four cycles of ABVD followed by adjuvant radio-therapy: analysis of efficacy and long-term toxicity. AB - BACKGROUND AND OBJECTIVES: The use of combined modality therapy in early-stage Hodgkin's disease can spare staging laparotomy and reduces the risk of relapse compared to radiation alone. This paper reports on the efficacy and long-term events of a combined modality approach consisting of a brief course of chemotherapy followed by adjuvant radiotherapy, without laparotomy, in early stage Hodgkin's disease. DESIGN AND METHODS: This study included 78 patients with Hodgkin's disease (20 in stage I and 58 in stage II); 60% had mediastinal enlargement (12% had bulky disease) and 5% had subdiaphragmatic disease. Their median age was 33 years (range: 15-64) and median follow-up 60 months. The treatment program consisted of four cycles of ABVD followed by adjuvant radiation to involved sites (43 patients) or involved and contiguous sites of disease (35 patients); radiation doses ranged from 30 to 36 Gy to uninvolved and involved sites, respectively; bulky disease received up to 44 Gy. Gonadal function in women was assessed by hormonal tests and evaluation of menses; young men were given the opportunity to have their semen cryopreserved. RESULTS: The treatment program was completed in a median of 6.2 months (range: 5-10). The complete remission rate was 88% after 4 courses of ABVD and 98.7% after adjunctive RT. The 5-year relapse-free survival was 97% and overall survival 98%; three patients died, one of disease progression and two of small cell lung carcinoma. Long-term events included three cases of pulmonary fibrosis with symptomatic interstitial disease, one case of dilated cardiomyopathy with cardiac failure (all had received mediastinal radiation) and four cases of dysthyroidism. Fertility was preserved in young women, with three subsequent normal pregnancies. Second neoplasms included two small cell lung carcinomas and one breast carcinoma. INTERPRETATION AND CONCLUSIONS: In early-stage Hodgkin's disease, four cycles of ABVD followed by adjuvant radiotherapy produced a 5-year overall survival of 98%. Prolonged monitoring for therapy-related long term complications is mandatory in these potentially curable patients. PMID- 11025595 TI - An ELISA system to detect anti-factor VIII antibodies without interference by lupus anticoagulants. Preliminary data in hemophilia A patients. AB - BACKGROUND AND OBJECTIVES: Difficulties in identifying the coexistence of neutralizing anti-factor VIII antibodies (anti-fVIII) and lupus anticoagulant (LA) are mainly due to the interference of LA on anti-fVIII assays. Our aim was to reveal the presence of anti-fVIII using a system that is not affected by LA. DESIGN AND METHODS: We developed an enzyme-linked immunosorbent assay (ELISA) method that uses phospholipid-free recombinant factor VIII as the antigen. A monoclonal anti-fVIII was tested as a positive control, excluding non-specific binding by using two unrelated monoclonal antibodies. The ELISA was performed on hemophilic plasmas with anti-fVIII and negative LA (n=12) or without inhibitors (n=12). Two hemophilic plasmas with LA and presumably anti-fVIII were also assayed. Positive LA (n=12) and normal (n=10) plasmas were tested as negative controls. RESULTS: All (12/12) plasmas with anti-fVIII and 5/12 hemophilic plasmas without inhibitors were positive; LA and normal plasma controls were negative. INTERPRETATION AND CONCLUSIONS: Results presented here show that LA does not interfere with the anti-fVIII ELISA: However, the assay detects both neutralizing and non-neutralizing anti-fVIII antibodies, therefore a neutralizing effect must be confirmed through functional tests. PMID- 11025596 TI - The influence of age, sex, vitamin B(12), folate levels and methylenetetrahydrofolate reductase C677T genetic mutations on plasma homocysteine in the Chinese population. AB - BACKGROUND AND OBJECTIVES: Thromboembolic diseases remain a major cause of morbidity and mortality in most countries. The present study was thus conducted to determine the influences of age, sex, the methylenetetrahydrofolate reductase (MTHFR) gene mutation and the B vitamins on the plasma homocysteine (Hcy) levels in the Chinese. Our previous study found that Chinese carry the same mutation of the methylenetetrahydrofolate reductase (MTHFR) gene described in Western populations, with a 677CAET substitution being another possible cause of thrombosis. DESIGN AND METHODS: The study population comprised 445 consecutively enrolled Chinese subjects of different ages and sex. Overall 69 subjects were found to have homozygous 677CAET mutation of the MTHFR gene, and were classified as Group I; 164 subjects were found to have heterozygous mutation and classified as Group II; 212 had no such mutation and were classified as Group III. RESULTS: The mean plasma Hcy did not differ significantly between these 3 groups. When each group was divided again by gender, we found that both age and plasma Hcy levels were significantly higher in the males than in the females. In addition to Hcy levels, we also measured plasma vitamin B(12) and folate levels in 258 randomized subjects. Univariate and multivariate analysis showed MTHFR mutation could affect Hcy level, and univariate and multivariate analysis showed that age, MTHFR mutation and vitamin B(12) could affect the log(Hcy) levels. INTERPRETATION AND CONCLUSIONS: We demonstrate that some Chinese carry the 677CAET mutation of the methylenetetrahydrofolate reductase gene. This could affect their homocysteine levels and thus be a risk factor for thromboembolic disease. PMID- 11025597 TI - Blood donors with 'medium' or 'minor' risk factors for human immunodeficiency virus infection: are they eligible for donation? AB - BACKGROUND AND OBJECTIVES: We conducted a longitudinal prospective study to assess the eligibility to blood donation of donors with 'minor' risk factors (i.e. minor surgery, professional exposure, cohabitation with 'high risk' people, occasional use of light drugs) or 'medium' risk factors for human immuno deficiency virus (HIV) infection (i.e. casual sexual relationship, multiple heterosexual exposure, sexual partnership with subjects at risk, regular use of light drugs). DESIGN AND METHODS: During a 4-year period we administered a psychosocial questionnaire to all donors attending our Center. In addition we determined anti-HIV, anti-hepatitis C virus (HCV), hepatitis B surface antigen (HBsAg) and syphilis serology (TPHA) at entry to the study and at 6-month intervals. RESULTS: Of 25,367 donors, 1,535 (6%) reported medium and 8,761 (34%) minor risk. At enrollment into the study, 4 medium risk donors were anti-HIV positive and there was a significantly higher rate of positivity for TPHA (0.33% vs 0.07%) and anti-HCV (1.17% vs 0.63%) in this group than in donors reporting no risk. No anti-HIV positivity was observed in minor or no risk donors. During a median follow-up of 18 months, none of 24,404 donors undergoing 106,503 visits seroconverted to HIV. The incidences of novel HCV and syphilis infections were almost one log greater in donors at medium risk (3 and 1x10-4/yr, respectively) than in no risk donors (0.4 and 0.1x10-4/yr, respectively). Medium risk donors were more frequently males (Odds Ratio=3.2, 95% confidence interval= 2.8-3.7), aged 26-35 yrs (1.52; 1.3-1.78), single (1.4; 1.2-1.6), divorced (2; 1.4-2.8), freelance workers (1.43; 1.1-1.9) and first-time donors (1.8; 1.6-2.1) than no risk donors. INTERPRETATION AND CONCLUSIONS: The only 4 HIV positive subjects of the cohort were medium risk donors picked up at enrollment. No HIV seroconversion was observed during the study. On the basis of this study we will continue to defer 'medium' risk donors. PMID- 11025598 TI - Hematopoietic cell transplantation in acute lymphoblastic leukemia: better long term event-free survival with conditioning regimens containing total body irradiation. AB - BACKGROUND AND OBJECTIVES: There is only limited experience with conditioning regimens based on busulfan for patients with acute lymphoblastic leukemia (ALL). Therefore, the aim of this study was to compare the event-free survival (EFS), transplant-related mortality (TRM) and the probability of relapse (PR) of patients undergoing hematopoietic cell transplantation (HCT) for ALL conditioned with or without total body irradiation (TBI). DESIGN AND METHODS: The study sample consisted of 156 patients conditioned with regimens based on TBI (n=114) or on high doses of oral busulfan (BU) (n=42). Most of the BU group received phenytoin as prophylaxis for seizures. The median follow-up was 6 years. RESULTS: EFS at 6 years was 43% (95% CI 35%-51%) versus 22% (95% CI 10%-34%) in the TBI and BU subsets respectively (p=0.01). TRM at 18 months was 22% and 17% in the BU and TBI groups (p=0.24), respectively. At 3 years actuarial PR was 71% in the BU group and 47% in the TBI group (p=0.01). In the multivariable analysis, a worse EFS was associated with BU, relative risk (RR) 1.7; advanced disease versus 1st and 2nd complete remission (CR) at HCT, RR 2.5; absence of chronic graft-versus host disease, RR 1.8; development of veno-occlusive disease RR 2.2 and shorter CR duration before transplant. INTERPRETATION AND CONCLUSIONS. TBI was associated with a lower relapse rate and better EFS, even in patients in 1(st )and 2(nd) CR, than schemes based on high doses of busulfan. This suggests that conditioning regimens based on TBI should remain the standard method of preparative regimen for patients with ALL. PMID- 11025600 TI - Molecular basis for therapeutic decisions in chronic myeloid leukemia patients after allogeneic bone marrow transplantation. AB - BACKGROUND AND OBJECTIVES: Recent progress in the development of diagnostic techniques has greatly facilitated the monitoring of minimal residual disease (MRD) in patients with chronic myeloid leukemia (CML) after allogeneic bone marrow transplantation (BMT), the only curative treatment for this disease. The presence of the P210(bcr-abl) rearrangement in CML cells has allowed highly sensitive detection of MRD by polymerase chain reaction (PCR). However, complete eradication of the leukemic clone may not be a necessary prerequisite for long term remission or cure. This observation limits the value of qualitative PCR analysis for prediction of progressive disease and highlights the need to monitor the proliferative activity of the malignant clone in order to permit timely detection of impending relapse and, thus, early therapy. This article discusses the applicability of several molecular methods to the monitoring of treatment efficacy and early assessment of clonal expansion in patients with CML after BMT. It also presents guidelines for clinical use of PCR analyses and the most effective approaches to treat relapsed patients. INFORMATION SOURCES: The authors have been working in this field, both experimentally and at a clinical level, contributing original papers to peer-reviewed journals. The material examined in this review includes articles published in journals covered by MedLine and reviews from journals with a high impact factor. STATE OF THE ART AND PERSPECTIVES: In view of the very limited value of qualitative PCR in detecting CML patients destined to relapse after BMT, several investigators have developed molecular assays that enable the kinetics of MRD to be monitored over time (e.g. quantitative PCR for P210(bcr-abl), PCR analysis of whole blood/lineage-specific chimerism and qualitative PCR for P190(bcr-abl)). These molecular strategies closely trace the kinetics of leukemic regrowth. Disease evolution in relapsed patients is consistently characterized by the sequential detection of increasing P210(bcr-abl) transcript levels, increasing myeloid mixed chimerism and finally, P190(bcr-abl) positivity preceding cytogenetic relapse. A 10-fold or greater increase in the expression of P210(bcr-abl) confirmed by a minimum of three independent quantitative PCR analyses and/or a progressive increase in the percentage of host myeloid cells in three consecutive chimerism analyses and/or P190(bcr-abl) mRNA detection must be regarded as an indication of incipient disease progression and should provide a rationale for initiation of treatment. There are various approaches to the management of the patient who relapses. The first step, if possible, is to reduce or terminate immune suppression. If the patient is not receiving this therapy, he or she can be treated with hydroxyurea or interferon or can be offered a second transplant. However, infusion to the patient of lymphoid cells (DLI) collected from the original donor has the capacity to restore complete remission in 70-80% of cases. Currently, several strategies are being used to minimize the severity of graft-versus-host disease after DLI (optimization of transfused lymphocyte doses, modification of the transfused lymphocyte subsets, administration of lymphocytes in escalating doses or lymphocyte transfection with a suicide gene), to reduce the incidence of marrow aplasia (stem cell support) and to increase the rate of complete responses (cytokines associated with DLI, leukemia-reactive cytotoxic lymphocytes, tyrosine kinase inhibitors or pre-emptive DLI). PMID- 11025599 TI - Mucormycosis in hematologic malignancies: an emerging fungal infection. AB - BACKGROUND AND OBJECTIVES: In recent years pulmonary mucormycosis has been reported in patients with leukemia and lymphoma and bone marrow transplant recipients. It carries an extremely poor prognosis. We report our experience of clinical findings, diagnostic procedures, treatment and outcome of mucormycosis diagnosed in neutropenic patients affected by hematologic neoplasms admitted to our Department. DESIGN AND METHODS: From November 1987 to July 1999 we observed 13 cases of Mucor. Their median age was 61 years (range 20-75), and they were predominantly in the aplastic post-chemotherapy period (12/13), affected by acute myeloid leukemia (11 cases ) or non-Hodgkin's lymphoma (2 cases). Six patients (all with leukemia) were receiving inductionEth consolidation therapy, 7 had progressive hematologic disease. At the onset of infection all patients were neutropenic (N < 0.5x10(9)/L). No patients had diabetes mellitus. Two patients had been receiving steroid therapy for 5 and 7 days. RESULTS: The lung was involved in all cases (13/13); disseminated disease was present in 8/13 patients. All cultures (blood, sputum, nasal swabs and bronchoalveolar lavage) were negative. In 3 patients a histologic diagnosis was made in vivo: in 1 patient by percutaneous pulmonary biopsy, in 1 patient by pulmonary lobectomy, and in the last patient by percutaneous pulmonary biopsy confirmed by excision of a cerebellar abscess. In the remaining 10 cases diagnosis was made post-mortem. Five patients were not treated, 2 because of poor clinical condition and 3 because fungal infection was not suspected. Amphotericin B (1 mg/kg/day) was given empirically to 6 patients and 2 responded to treatment. The remaining 2 patients with neurologic symptoms at the onset of infection were treated with liposomal amphotericin, Ambisome, one with 3 and one with 5 mg/kg/day; of these two patients the first died in 4 days; the second, with both pulmonary and cerebellar localizations, was treated successfully with 5 mg/kg/day for 4 weeks and then with 3 mg/kg/day, and excision of a brain abscess at neutrophil recovery (total dose of Ambisome: 12,000 mg). The 3 surviving leukemic patients were able to complete subsequent consolidation therapy using amphotericin B or liposomal amphotericin as secondary prophylaxis during aplasia. INTERPRETATION AND CONCLUSIONS: In neutropenic hematologic patients Mucor is rarely suspected. In our patients infection was often characterized by disseminated disease and a rapidly fatal course; only early aggressive amphotericin B (or Ambisome) treatment together with neutrophil recovery appeared to improve the outcome. Diagnosis is very important for programming antifungal therapy and secondary prophylaxis with amphotericin B, because Mucor is usually resistant to itraconazole. PMID- 11025601 TI - Leukemic relapse as T-acute lymphoblastic leukemia in a patient with acute myeloid leukemia and a minor T-cell clone at diagnosis. AB - BACKGROUND AND OBJECTIVES: Lineage classification needs to be established before starting chemotherapy in acute leukemias. In some cases, mixed populations can be found and these are differentiated by antigenic expression patterns. DESIGN AND METHODS: We report the case of a patient with acute myelogenous leukemia whose relapse was classified as T-acute lymphoblastic leukemia (T-ALL). RESULTS: Flow cytometry analysis at diagnosis enabled us to identify a minor T-cell subclone which progressively increased and became dominant at relapse. There were no changes at cytogenetic and molecular levels. INTERPRETATION AND CONCLUSIONS: This case illustrates the usefulness of multiparametric flow cytometry for assessing minor leukemic populations. PMID- 11025602 TI - Translocation (8;16) in a patient with acute myelomonocytic leukemia, occurring after treatment with fludarabine for a low-grade non-Hodgkin's lymphoma. AB - We describe a 65-year old woman who developed a t(8;16)(p11;p13) positive acute myeloid leukemia (AML)-M4 without a prior myelodysplasia thirty-six months after a low-grade non-Hodgkin's lymphoma treated with alkylating agents (chlorambucil and cyclophosphamide) and fludarabine, a purine analog with a significant activity in lymphoproliferative disorders. The t(8;16)(p11;p13) is present in 0.4% of AML of M4-M5 cytotype. In the present case it was identified by conventional cytogenetics; involvement of the MOZ and CBP genes was demonstrated by fluorescence in situ hybridization, but not by reverse transcription polymerase chain reaction. The patient died of sepsis after the first course of induction chemotherapy. This is the first t(8;16) AML-M4 arising after fludarabine treatment of which the leukemogenic role in our case is very difficult to ascertain. Most t(8;16) t-AML cases had received anthracyclines with or without cyclophosphamide; none was ever administered chlorambucil. Our patient was never given anthracyclines and the cumulative doses of chlorambucil and cyclophosphamide employed were low. PMID- 11025603 TI - Haemophilia B in a female caused by skewed inactivation of the normal X chromosome. AB - Hemophilia B (factor IX deficiency) is an X-linked recessive disorder with a prevalence of 1:30,000 male births, which rarely affects females. A missense mutation T38R (6488C>G) of the factor IX (FIX) gene was characterized in a young female with moderate-to-severe hemophilia B. She is heterozygous for this mutation, which she inherited from her carrier mother. Analysis of the methyl sensitive HpaII sites in the first exon of the human androgen-receptor locus indicated a de novo skewed X-chromosomal inactivation. This indicates that the paternal X-chromosome carrying her normal FIX gene is the inactive one, which has led to the phenotypic expression of hemophilia B in this patient. PMID- 11025604 TI - Breast cancer or cutaneous lymphoma? PMID- 11025605 TI - Bronchiolocentric pulmonary involvement due to chronic lymphocytic leukemia. PMID- 11025606 TI - Staphilococcus contamination of blood sample mimicking platelet clumps. PMID- 11025607 TI - Acute myeloblastic leukemia with minimal myeloid differentiation featuring a three-way translocation t(8;13;14) PMID- 11025608 TI - Monitoring of minimal residual disease and mixed chimerism in a case of high-risk TEL/AML1+ acute lymphoblastic leukemia pre- and post-bone marrow transplantation. Czech Pediatric Haematology Working Group. PMID- 11025609 TI - Use of molecular techniques to confirm true re-emergence of the original clone and to track minimal residual disease in a case of late extramedullary relapse of childhood acute lymphoblastic leukemia. PMID- 11025610 TI - Cryptococcal meningitis during front-line chemotherapy for acute lymphoblastic leukemia. PMID- 11025611 TI - Chromosomal instability in chronic myeloid leukemia: Philadelphia breakpoints are irrespective to spontaneous breakage and fragile sites. PMID- 11025612 TI - Vascular endothelial growth factor isoforms 121 and 165 are expressed on B chronic lymphocytic leukemia cells. PMID- 11025613 TI - Expansion of CD3+CD56+ cytotoxic cells from patients with chronic lymphocytic leukemia: in vitro efficacy. PMID- 11025614 TI - Non-ALC peripheral T-cell lymphomas in children: report on two cases and review of the literature. PMID- 11025616 TI - Feasibility and efficacy of high-dose etoposide followed by low-dose G-CSF as a mobilization regimen in patients with non-Hodgkin's lymphoma. PMID- 11025615 TI - Low-dose of thalidomide in the treatment of refractory myeloma. PMID- 11025617 TI - Disappearance of FVIII inhibitors in a severe hemophilia A neonate after steroid treatment correlated with a cytokine shift toward a T-helper 2 pattern. PMID- 11025618 TI - Simultaneous occurrence of Clostridium difficile and Cytomegalovirus colitis in a recipient of autologous stem cell transplantation. PMID- 11025619 TI - Expression of CD34 by megakaryocytes in myelodysplastic syndromes. PMID- 11025620 TI - Kinetics variation of CD34+ and CD34+CD90+ in subjects following different mobilizing protocols. PMID- 11025621 TI - The decade of the brain: a brief review. AB - Recognising the huge burden of neurological and psychiatric disorders and prompted by the potentials of new techniques of molecular biology, biotechnology, genetics and imaging to study these, the 1990s were declared the 'decade of the brain'. This stimulated global scientific efforts to understand the human brain in health and disease. This review summarises some of the major research achievements during the decade. While it is impossible to provide a comprehensive summary of the voluminous data that has been generated, it was decided to provide a bird's eye view of the recent advances in the fields of developmental neurobiology, neurogenetics, neurochemistry and imaging of the brain, which have direct relevance for the clinicians. PMID- 11025622 TI - Reconstruction of the skull base: a review of personal techniques. AB - With increasing technological and surgical sophistication in cranial base and craniofacial surgery, reconstructive efforts are challenged to provide a reliable means of compartmentalisation. Some improvised methods of basal reconstruction with vascularised pedicle flap are described in this presentation. The dependable blood supply, long length, ease of harvesting and the ability to alter the arc of rotation of the described flaps make them versatile for lining skull base. The flaps may be used to cover defects, may be folded for bulk and may be used to carry blood supply to poorly vascularised recipient sites. The techniques are presented. PMID- 11025623 TI - Ideal ward round making in neurosurgical practice. AB - The success of a perfect ward round lies in the role of the consultant leading the 'round making group' (RMG) as well as the hallmark of effective questioning and participation of each member. Twelve senior consultants with more than 10 years' experience in neurosurgical practice at three different university hospitals were observed during round making by a participant observer. Observations were made on the group climate of the RMG, the leadership pattern and language expressed by the clinician conducting the round and the effectiveness in his performance as a leader during clinical discussions. The group climate showed evidence of good productivity and flexibility with 92% and 75% consultants, pleasantness of climate was above average with only 50% (6/12) and poor objectivity with 42% (5/12) consultants. Forty two percent of the consultants were not always very well comprehensible, while only 50% (6/12) spoke exactly fitting the occasion. Only 33% (4/12) of the consultants used humour effectively, while 42% (5/12) spoke unnecessarily in between discussion and were poor in introducing the problems of patient to the round making group. Ward round making in neurosurgical practice needs a holistic approach with motivation, planning, leadership skills and structured curriculum to fulfill its objectives. PMID- 11025624 TI - A double blind controlled study of propranolol and cyproheptadine in migraine prophylaxis. AB - Role of propranolol and cyproheptadine in the prophylaxis of migraine was studied in a controlled double blind trial. Two hundred fifty-nine patients were divided into four groups. Each group was either given a placebo, cyproheptadine, propranolol or a combination of the latter two drugs. The patients were followed for a period of three months. Significant relief in frequency, duration and severity from migranous attacks was seen in all drug treated groups over placebo. Significant correlation in response was seen in frequency, duration and severity in all the groups which received drugs. Statistically more significant relief was seen in cyproheptadine and propranolol treated group as compared to individual drug treated groups. In cyproheptadine and propranolol treated groups, the dropout rate was lower and associated symptoms were better relieved than in other groups. The study shows efficacy of combination of cyproheptadine and propranolol in migraine prophylaxis. PMID- 11025625 TI - Effect of dichloracetate on infarct size in a primate model of focal cerebral ischaemia. AB - Acidosis is a major contributing factor towards spread of the ischaemic focus in the brain. Drugs that increase pyruvate dehydrogenase activity could decrease the formation of lactic acidosis. The sodium salt of dichloracetic acid (DCA) has been found to be effective in reducing lactate. This study was undertaken to study the efficacy of DCA in reducing infarct size in experimental focal ischaemia in monkeys. Macaca radiata monkeys in the treatment group were given 35 mg per kilogram of dichloracetate intravenously immediately before occluding and interrupting the middle cerebral artery, and the control group was given saline as placebo under similar conditions. Mean infarct size expressed as a percentage of the size of the hemisphere in all the three brain slices was 35.38 in the control group as against l2.06 in the treated group (p=0. 0008). PMID- 11025627 TI - P300 in newly diagnosed non-dementing Parkinson's disease: effect of dopaminergic drugs. AB - Changes in cognitive function are an integral part of the clinical presentation of Parkinson's Disease (PD). P300 potential studies in early stages of Parkinson's disease are lacking and effect of L-dopa therapy on these potentials is controversial. In this study, changes in P300 potentials in early stages of PD and effects of dopaminergic therapy were investigated. P300 waves were elicited by standard auditory 'odd ball' paradigm and were recorded before the start of therapy and 15 days, 3 and 6 months after the start of L-dopa therapy in 25 newly diagnosed patients with idiopathic PD. All patients were classified according to Hoehn and Yahr scale. Minimental status examination (MMSE) was done in all. Control group had 20 normal subjects. The P300 latency was not significantly increased in early Parkinson's disease. This latency was reduced with dopaminergic therapy on 15th day, but increased later. Implications of the data are discussed. PMID- 11025628 TI - Cervical spinal cord injury without radiological abnormality in adults. AB - Spinal cord injury occurring without concomitant radiologically demonstrable trauma to the skeletal elements of the spinal canal rim, or compromise of the spinal canal rim without fracture, is a rare event. Though documented in children, the injury is not very well reported in adults. We present seventeen adult patients with spinal cord injury without accompanying fracture of the spinal canal rim, or vertebral dislocation, seen over seven years. None had preexisting spinal canal stenosis or cervical spondylosis. Following trauma, these patients had weakness of all four limbs. They were evaluated by MRI (CT scan in one patient), which showed hypo / isointense lesion in the cord on T1 weighted images, and hyperintensity on T2 weighted images, suggesting cord contusion or oedema. MRI was normal in two patients. With conservative management, fifteen patients showed neurological improvement, one remained quadriplegic and one died. With increasing use of MRI in the evaluation of traumatic myelopathy, such injuries will be diagnosed more often. The mechanism of injury is probably acute stretching of the cord as in flexion and torsional strain. Management is essentially conservative and prognosis is better than that seen in patients with fracture or dislocation of cervical spine. PMID- 11025626 TI - Tryptophan and tyrosine catabolic pattern in neuropsychiatric disorders. AB - Catabolism of tryptophan and tyrosine in relation to the isoprenoid pathway was studied in neurological and psychiatric disorders. The concentration of trytophan, quinolinic acid, kynurenic acid, serotonin and 5-hydroxyindoleacetic acid was found to be higher in the plasma of patients with all these disorders; while that of tyrosine, dopamine, epinephrine and norepinephrine was lower. There was increase in free fatty acids and decrease in albumin (factors modulating tryptophan transport) in the plasma of these patients. Concentration of digoxin, a modulator of amino acid transport, and the activity of HMG CoA reductase, which synthesizes digoxin, were higher in these patients; while RBC membrane Na+-K+ ATPase activity showed a decrease. Concentration of plasma ubiquinone (part of which is synthesised from tyrosine) and magnesium was also lower in these patients. No morphine could be detected in the plasma of these patients except in MS. On the other hand, strychnine and nicotine were detectable. These results indicate hypercatabolism of tryptophan and hypocatabolism of tyrosine in these disorders, which could be a consequence of the modulating effect of hypothalamic digoxin on amino acid transport. PMID- 11025629 TI - Laminoplasty: an evaluation of 24 cases. AB - Cervical expansive laminoplasty has been advocated as an alternative procedure to laminectomy for the decompression of the cervical spine. It provides favourable cord decompression and stabilisation of the cervical spine and is a simpler and safer alternative to anterior fusion and laminectomy for myelopathy and myeloradiculopathy, due to multisegmental cervical spondylosis and ossified posterior longitudinal ligament. We report our experience in 24 patients with this procedure, 12 of whom had myelopathy and another 12 had myeloradiculopathy. The earliest symptom to improve was radicular pain or paraesthesia (75%). A reduction in spasticity was seen in 21 of the 24 patients (87.5%). Eleven patients had improvement in their motor power during a follow up period ranging from 1 month to 14 months. One patient deteriorated following the procedure and developed Brown Sequard features due to under riding of the lamina on the hinged side, another had severe post operative paraesthesias, while one patient had a CT scan evidence of 'closing of the door', without being symptomatic for it. The technique of the procedure is discussed and the pertinent literature reviewed. PMID- 11025630 TI - Pre and post betahistine therapy 99m Tc - HMPAO brain spect studies in patients with vertigo. AB - Vertebro basilar insufficiency (VBI) is a well known cause of vertigo. Brain Single Photon Emission Computed Tomography (SPECT) is an important diagnostic tool to detect and to quantitate the perfusion abnormalities in different areas of the brain. Effect of an antivertigo drug Betahistine on improving the hypoperfusion in different areas of the brain in vertigo patients was studied using brain SPECT. Betahistine at a dose of 16 mg three times daily was shown to improve perfusion in the hypoperfused areas of the brain resulting in relief from symptoms of vertigo. The cerebellar region, which is the most important area involved in vertigo patients with vascular pathology, showed almost complete normalisation of perfusion following Betahistine therapy. PMID- 11025631 TI - Neurocysticercosis like presentation in a case of CNS tuberculosis. AB - A clinical picture consisting of seizures, multiple non-tender subcutaneous nodules, and multiple 'nodular or ring' enhancing lesions in computed tomography of the brain is considered characteristic of neurocysticercosis in an endemic area. 1,2 A case with a similar clinical picture, in whom serological tests and histopathological examination of subcutaneous nodule established tuberculosis as a cause, is presented. PMID- 11025632 TI - Hypoglycaemic neuropathy: a case report. AB - A 53 years old male, a known case of ankylosing spondylitis having recurrent attacks of hypoglycaemia, developed symmetrical distal sensorimotor neuropathy. The neuropathy was axonal with secondary demyelination. Evidence of vasculopathy was also noted on histopathology of the nerve. Serum C-peptide level was low, a feature reported with autoimmune hypoglycaemia with antireceptor antibodies. The patient showed spontaneous recovery. PMID- 11025633 TI - Congenital myasthenic syndrome: report of four cases and brief review of literature. AB - The term 'congenital myasthenic syndrome' (CMS) encompasses a number of heterogeneous disorders characterised by myasthenic symptoms since birth, usually with positive family history and absence of acetyl choline receptor antibodies. Recent advances in electrophysiology and ultrastructural analysis of neuromuscular junction have made it possible to identify the various defects underlying these disorders. We report four cases of CMS, with a review of literature. PMID- 11025634 TI - Brainstem abscess complicating tetralogy of Fallot successfully treated with antibiotics alone. AB - Medically treated brainstem abscess in a 11 year old boy with tetralogy of Fallot is reported. There was a complete resolution of the lesion without any neurologic sequelae during parenteral antibiotic therapy with crystalline penicillin, chloramphenicol and metronidazole. The pathogenesis and management of cardiogenic brain abscesses in general and brainstem abscess in particular has been reviewed. PMID- 11025635 TI - Mycotic aneurysm on posterior cerebral artery: resolution with medical therapy. AB - Mycotic aneurysms on the branches of vertebro basilar artery are rare. A patient of infective endocarditis with mycotic aneurysm on the posterior cerebral artery is described. The aneurysm resolved with medical therapy. Controversies regarding the management of mycotic aneurysms are discussed. PMID- 11025636 TI - Gluteal neuralgia - unusual presentation in an adult with intrasacral meningocele: a case report and review of literature. AB - A nineteen year old man with intrasacral meningocele is reported, who presented with long standing episodic gluteal pain and progressive muscle wasting. Magnetic resonance imaging established the diagnosis. Surgical excision relieved the pain but muscle wasting persisted. Pertinent literature is reviewed. PMID- 11025637 TI - Clinical, pathological and magnetic resonance imaging features of focal myositis: report of three cases. AB - Focal myositis is a rare idiopathic form of inflammatory myositis involving a single muscle. The presentation is often as a soft tissue pseudotumour. Magnetic resonance imaging (MRI) is a useful noninvasive imaging modality to demonstrate focal nature of the lesion. We describe three patients presenting as pseudotumour of the lower leg. MRI showed hyperintense signals in the involved muscle. PMID- 11025638 TI - Calcified falx meningioma. AB - A totally calcified mid third falcine meningioma in an elderly male patient is presented. An uneventful enmasse excision was performed. Advantages of positioning on ipsilateral side for paramedian extracerebral lesions are highlighted. PMID- 11025639 TI - Diagnostic dilemma in a case of early spinal tumour fracture (Plasmacytoma): a case report. AB - MRI findings of D12 fracture were suspicious of a pathological fracture. However, biopsy did not show any evidence of tumour. This resulted in the patient following up after one and half years with an epidural mass lesion and neurological deterioration. This paper highlights some of the MRI features of fractures associated with underlying pathology over traumatic or osteoporotic fracture. PMID- 11025640 TI - Paraganglioma of cauda equina: a case report. AB - Histopathologically and immunologically confirmed case of paraganglioma of cauda equina region is described. PMID- 11025641 TI - Cortical sinovenous thrombosis in a child with nephrotic syndrome and iron deficiency anaemia. AB - Cortical sinovenous thrombosis in a child with nephrotic syndrome and iron deficiency anaemia is described. The most probable mechanism for the hypercoagulable state was thrombocytosis associated with iron deficiency anaemia. The other possible contributing factor might have been the diuretic therapy during the phase of relapse. PMID- 11025642 TI - Spontaneous resolution of a non-functioning pituitary adenoma following an apoplexy. AB - A rare case of non-functioning pituitary adenoma, which completely resolved following an apoplectic event without producing hypopituitarism, is described. PMID- 11025643 TI - Cerebral venous thrombosis with cortical infarct: neuroimage. PMID- 11025644 TI - Particle irradiation induces FGF2 expression in normal human lens cells. AB - Particle Irradiation Induces FGF2 Expression in Normal Human Lens Cells. Particle radiations, including both proton and helium-ion beams, have been used to successfully treat choroidal melanoma, but with the complication of radiation induced cataract. We have investigated a role for radiation-induced changes in the expression of basic fibroblast growth factor (FGF2) gene expression as part of the mechanism(s) underlying lens cell injury associated with cataract. Normal human lens epithelial (HLE) cells were cultured in vitro on extracellular matrix (ECM) originated from bovine corneal endothelial cells. This study reports evidence for rapid but transient induction of FGF2 transcripts, an increase of between 5- and 8-fold, within 0.5 h after exposure to particle radiation, followed by another wave of increased transcription at 2-3 h postirradiation. Immunofluorescence results confirm the enhanced levels of FGF2 protein rapidly after exposure to protons or helium ions, followed by another wave of increased activity unique to helium at 6 h postirradiation. This second wave of increased immunoreactivity was not observed in the proton-irradiated samples. Total FGF2 protein analysis after helium-ion exposures shows induced expression of three FGF2 isoforms, with an increase of up to 2-fold in the 18-kDa low-molecular weight species. Studies of the effects of protons on individual FGF2 protein isoforms are in progress. Several mechanisms involving a role for FGF2 in radiation-induced cataract are discussed. PMID- 11025645 TI - Inactivation of aerobic and hypoxic cells from three different cell lines by accelerated (3)He-, (12)C- and (20)Ne-ion beams. AB - The LET-RBE spectra for cell killing for cultured mammalian cells exposed to accelerated heavy ions were investigated to design a spread-out Bragg peak beam for cancer therapy at HIMAC, National Institute of Radiological Sciences, Chiba, prior to clinical trials. Cells that originated from a human salivary gland tumor (HSG cells) as well as V79 and T1 cells were exposed to (3)He-, (12)C- and (20)Ne ion beams with an LET ranging from approximately 20-600 keV/micrometer under both aerobic and hypoxic conditions. Cell survival curves were fitted by equations from the linear-quadratic model and the target model to obtain survival parameters. RBE, OER, alpha and D(0) were analyzed as a function of LET. The RBE increased with LET, reaching a maximum at around 200 keV/micrometer, then decreased with a further increase in LET. Clear splits of the LET-RBE or -OER spectra were found among ion species and/or cell lines. At a given LET, the RBE value for (3)He ions was higher than that for the other ions. The position of the maximum RBE shifts to higher LET values for heavier ions. The OER value was 3 for X rays but started to decrease at an LET of around 50 keV/micrometer, passed below 2 at around 100 keV/micrometer, and then reached a minimum above 300 keV/micrometer, but the values remained greater than 1. The OER was significantly lower for (3)He ions than the others. PMID- 11025646 TI - The degree of pigmentation modulates the radiosensitivity of human melanoma cells. AB - The relationship between cell pigmentation and radiosensitivity was investigated in two selected human melanoma cell lines with different melanin content (mixed type: eumelanin and pheomelanin, and pheomelanotic phenotypes). The same study was also done after stimulation of melanogenesis (1) by addition of the melanin precursor l-tyrosine to each of the cell lines separately and (2) by irradiation alone with doses ranging from 0 to 10 Gy. We found that a decrease in cell radiosensitivity was correlated with the type of melanin, with a clear involvement of eumelanin rather than pheomelanin. Increasing the intracellular content of both melanins promoted the growth of irradiated cells. Moreover, at a dose of 10 Gy, both tyrosinase activity and melanin cell content were significantly increased in the absence of any other melanogenesis promoter. Our data suggest that the amount of intracellular melanin is inversely related to the radiosensitivity of melanoma cells and may explain at least in part the controversial responses to ionizing radiations reported for melanoma. PMID- 11025647 TI - Regulation of FOS by different compartmental stresses induced by low levels of ionizing radiation. AB - We irradiated different cellular compartments and measured changes in expression of the FOS gene at the mRNA and protein levels. [(3)H]Thymidine and tritiated water were used to irradiate the nucleus and the whole cell, respectively. (125)I Concanavalin A binding was used to irradiate the cell membrane differentially. Changes in FOS mRNA and protein levels were measured using semi-quantitative RT PCR and SDS-PAGE Western blotting, respectively. Irradiation of the nucleus or the whole cell at a dose rate of 0.075 Gy/h caused no change in the level of FOS mRNA expression, but modestly (1.5-fold) induced FOS protein after 0.5 h. Irradiation of the nucleus at a dose rate of 0.43 Gy/h induced FOS mRNA by 1.5 fold after 0.5 h, but there was no significant effect after whole-cell irradiation. FOS protein was transiently induced 2.5-fold above control levels 0.5 h after a 0. 43-Gy/h exposure of the nucleus or the whole cell. Irradiation of the cell membrane at a dose rate of 1.8 Gy/h for up to 2 h caused no change in the levels of expression of FOS mRNA or protein, but a dose rate of 6.8 Gy/h transiently increased the level of FOS mRNA 3-fold after 0.5 h. These data demonstrate the complexity of the cellular response to radiation-induced damage at low doses. The lack of quantitative agreement between the transcript and protein levels for FOS suggests a role for post-transcriptional regulation. PMID- 11025648 TI - Different patterns of DNA fragmentation and degradation of nuclear matrix proteins during apoptosis induced by radiation, hyperthermia or etoposide. AB - Several nuclear matrix proteins are substrates for proteolytic cleavage during apoptosis. Using Western blotting, the temporal patterns of cleavage of three nuclear matrix proteins (lamin B, NUMA and the nucleoporin TPR) were compared in HL60 cells induced to undergo apoptosis after irradiation, heat shock or treatment with etoposide. Flow cytometry was used to compare the kinetics of post cleavage degradation of lamin B, NUMA and TPR after irradiation, and to correlate DNA fragmentation with protein degradation in cells induced to undergo apoptosis with different agents. During radiation-induced apoptosis, cleavage and subsequent degradation of lamin B, NUMA and TPR occurred with different kinetics. Low-molecular-weight DNA fragmentation occurred subsequent to the initiation of NUMA cleavage, coincided with lamin B cleavage, but occurred before more extensive degradation of lamin B and NUMA. A similar sequence was observed for cells treated with etoposide. However, during heat-induced apoptosis, cleavage of lamin B and NUMA occurred much sooner compared to other agents, with NUMA cleaved into multiple fragments within 15 min after heating. We conclude that the hierarchical sequence and kinetics of degradative events contributing to nuclear disassembly during apoptosis are highly dependent on the inducing agent. Furthermore, the nuclear pore complex, like the nuclear lamina and internal nuclear matrix, is a target for proteolytic cleavage. PMID- 11025649 TI - Late effects of ionizing radiation on the microvascular networks in normal tissue. AB - Damage to the microvascular networks constitutes one of the most important components of ionizing radiation damage to normal tissue. Previously, we have reported the early (3, 7 and 30 days postirradiation) effects of ionizing radiation on the structure and function of normal tissue microvascular networks. Here we report on the late effects of ionizing radiation on the structural and functional changes in microvascular networks in locally irradiated (single 10-Gy dose) hamster cremaster muscles observed 60, 120 and 180 days postirradiation; age-matched animals were used as controls. As in the previous study, intravital microscopy was used to measure structural and functional parameters in complete microvascular networks in vivo. A factorial design was used to examine the effects of radiation status, time postirradiation, and network vessel type on the structure and function of microvascular networks. Our results indicate that the progression of radiation-induced microvascular damage continues during the late times but that there is partial recovery from radiation damage within 6 months postirradiation. Red blood cell flux, red blood cell velocity, and capillary blood flow in irradiated networks at 180 days postirradiation were significantly greater than control levels. As at the early times, all vessel types were not damaged equally by radiation at every time. PMID- 11025650 TI - Tissue damage after single high-dose intraoperative irradiation of the canine liver: evaluation in time by means of radionuclide imaging and light microscopy. AB - To establish the tolerance of liver tissue to single high-dose intraoperative irradiation, the histopathological changes in the canine liver after single high dose intraoperative irradiation were investigated by means of radionuclide imaging and light microscopy. Intraoperative irradiation at doses of 0, 10, 20, 25 or 30 Gy was applied to a part of the liver of 25 beagles. Radionuclide imaging using (99m)Tc-sulfur colloid was performed at several times during follow up. Elective humane killing was done 3 months and 1, 2, 3 and 5 years after irradiation. Light microscopy was used to identify histopathological alterations. There was no morbidity or mortality during a maximal follow-up of 5 years. In 40% of the animals, a region of diminished uptake was observed at the irradiation site. The regions of diminished uptake of the radiopharmaceutical agent became smaller with time. Light microscopic examination revealed severe parenchymal fibrosis, liver cell atrophy, and bile duct proliferation at the irradiated area 1 to 2 years after irradiation. At 3 and 5 years, vascular changes with endothelial proliferation and focal arteriolar hyalinosis were observed. This study demonstrates that intraoperative irradiation of a part of the liver in the canine model can be applied safely. Light microscopy confirmed that histological damage was not always accompanied by diminished uptake of the radiopharmaceutical agent at the irradiation site. PMID- 11025651 TI - Optical sensor-based oxygen tension measurements correspond with hypoxia marker binding in three human tumor xenograft lines. AB - Hypoxia has a negative effect on the outcome of radiotherapy and surgery and is also related to an increased incidence of distant metastasis. In this study, tumor pO(2) measurements using a newly developed time-resolved luminescence-based optical sensor (OxyLitetrade mark) were compared with bioreductive hypoxia marker binding (pimonidazole). Single pO(2) measurements per tumor were compared to hypoxia marker binding in tissue sections using image analysis. Both assays were performed in the same tumors of three human tumor lines grown as xenografts. Both assays demonstrated statistically significant differences in the oxygenation status of the three tumor lines. There was also a good correlation between hypoxia marker binding and the pO(2) measurements with the OxyLitetrade mark device. A limitation of the OxyLitetrade mark system is that it is not yet suited for sampling multiple sites in one tumor. An important strength is that continuous measurements can be taken at the same position and dynamic information on the oxygenation status of tumors can be obtained. The high spatial resolution of the hypoxia marker binding method can complement the limitations of the OxyLitetrade mark system. In the future, a bioreductive hypoxic cell marker for global assessment of tumor hypoxia may be combined with analysis of temporal changes in pO(2) with the OxyLitetrade mark to study the effects of oxygenation modifying treatment on an individual basis. PMID- 11025652 TI - Electron and photon spectra for three gadolinium-based cancer therapy approaches. AB - Some recent neutron capture therapy research has focused on using compounds containing the element gadolinium, which produces internal conversion and Auger cascade electrons. The low-energy, short-range Auger electrons are absorbed locally and increase cell killing dramatically as the gadolinium compounds are introduced into the cell nucleus and bind to the DNA. Detailed electron and photon spectra are needed for biophysical modeling and Monte Carlo calculations of damage to DNA. This paper presents calculated electron and photon spectra for three cases: thermal neutron absorption by (157)Gd, the beta-particle decay of (159)Gd, and the K-shell photoelectric event in gadolinium. The Monte Carlo sampling of atomic and nuclear transitions for each of the three cases was used to calculate a large number of representative decays. The sampled decays were used to determine average emissions and energy deposited in small spheres of tissue. The kinetic energy nuclear recoil from gamma-ray and electron emissions was calculated and found to be more than 10 eV for 26% of all (157)Gd neutron capture reactions. PMID- 11025653 TI - Measurement and validation of benchmark-quality thick-target tungsten X-ray spectra below 150 kVp. AB - Pulse-height distributions of two constant potential X-ray tubes with fixed anode tungsten targets were measured and unfolded. The measurements employed quantitative alignment of the beam, the use of two different semiconductor detectors (high-purity germanium and cadmium-zinc-telluride), two different ion chamber systems with beam-specific calibration factors, and various filter and tube potential combinations. Monte Carlo response matrices were generated for each detector for unfolding the pulse-height distributions into spectra incident on the detectors. These response matrices were validated for the low error bars assigned to the data. A significant aspect of the validation of spectra, and a detailed characterization of the X-ray tubes, involved measuring filtered and unfiltered beams at multiple tube potentials (30-150 kVp). Full corrections to ion chamber readings were employed to convert normalized fluence spectra into absolute fluence spectra. The characterization of fixed anode pitting and its dominance over exit window plating and/or detector dead layer was determined. An Appendix of tabulated benchmark spectra with assigned error ranges was developed for future reference. PMID- 11025654 TI - Unique response of double-stranded oligonucleotides containing a single 8-oxo-7,8 dihydroguanine to gamma rays in the frozen aqueous state at 77 K. AB - Two kinds of double-stranded oligonucleotides containing a single 8-oxo-7,8 dihydroguanine were labeled with (32)P at their 5' ends and exposed to gamma rays in the frozen aqueous state at 77 K, where both direct and quasi-direct effects of ionizing radiation predominate. Analysis of the oligonucleotides with 20% denaturing polyacrylamide gel electrophoresis revealed no difference in the immediate induction of strand breaks between oligonucleotides containing 8-oxo 7,8-dihydroguanine and their corresponding oligonucleotides with normal guanine, but piperidine-sensitive damage was induced more frequently in the former than in the latter. Sequence analysis of irradiated oligonucleotides showed that not only 8-oxo-7,8-dihydroguanine but also its neighboring bases and the cytosine residue that is paired to it became piperidine-sensitive in both oligonucleotides. These results suggest that 8-oxo-7,8-dihydroguanine, its neighboring bases and the opposite cytosine are candidates for radiation damage hot spots. PMID- 11025655 TI - Lack of RB protein correlates with increased sensitivity to UV-radiation-induced apoptosis in human breast cancer cells. AB - The underlying causes for different apoptotic responses in neoplastic cells are still not fully understood. We demonstrate here that a human breast cancer cell line, MDA-MB-468, which lacks the retinoblastoma protein (RB), is particularly sensitive to low doses of ultraviolet (UV) radiation. These cells are 15-20-fold more sensitive to UV radiation than RB-positive cell lines, as measured by both apoptosis and clonogenic assays. In addition, a prostate cancer cell line that lacks functional RB, DU-145, was found to have a similar apoptotic response to low doses of UV radiation. Based on these data, we hypothesized that the lack of RB is responsible for the extreme sensitivity of these cells to UV-radiation induced apoptosis. To further examine the role of RB in apoptosis, cells of RB positive human breast cancer and normal cell lines were infected with the human papilloma virus type 16 (HPV-16) E7 and assessed for UV-radiation sensitivity. The HPV-16 E7 protein is known to decrease levels of free RB in cells. Infection of RB-positive human breast cancer or normal cells with E7 resulted in a 4-5-fold increase in sensitivity to UV radiation compared to controls. The above data suggest a role for the RB protein in protecting cells from undergoing apoptosis in response to UV radiation. PMID- 11025656 TI - A new model of thermal inactivation and its application to clonogenic survival data for WiDr human colonic adenocarcinoma cells. AB - Based on the analysis of clonogenic survival data for human colonic adenocarcinoma cells (WiDr) after a single heating, a new model is proposed to describe cell survival after hyperthermia quantitatively. The effects of heat are explained as heat-induced cell damage assuming a first-order (single-hit) and a second-order (cumulative damage) process. Thus cell survival at a specified temperature can be described by the linear-quadratic (LQ) model. The proposed model is based on an alternative definition of the (single) thermal dose, given as the (normalized) product of heating time and a specified nonlinear function of the increase in temperature (relative to a threshold temperature) to be interpreted as the thermal dose rate. In further analogy to the modeling of the effects of low-dose-rate radiation, an inherent capacity of the cells to repair sublethal damage is assumed, and these effects are quantified by the usual g factor measuring incomplete repair effects. The model defines thermal dose response and isoeffect dose relationships, enabling a direct (i. e. single-step) analysis of the available thermal response data. Additionally, the analysis of our data based on heating times in the range from 0 to 360 min and temperatures from 41 to 46 degrees C and covering a broad spectrum of different densities of cells seeded for colony formation did not yield any evidence of the existence of a breaking point usually derived from Arrhenius plots based on the single-hit, multitarget model and the Arrhenius equation. The model includes no specific assumptions describing the development of thermotolerance, which can be assumed to be negligible under our experimental conditions. The proposed thermal dose response model correlates satisfactorily with the in vitro survival data for WiDr adenocarcinoma cells. PMID- 11025657 TI - Chemical genetic analysis of the budding-yeast p21-activated kinase Cla4p. AB - The p21-activated kinases (PAKs) are effectors for the Rho-family GTPase Cdc42p. Here we define the in vivo function of the kinase activity of the budding yeast PAK Cla4p, using cla4 alleles that are specifically inhibited by a cell-permeable compound that does not inhibit the wild-type kinase. CLA4 kinase inhibition in cells lacking the partially redundant PAK Ste20p causes reversible SWE1-dependent cell-cycle arrest and gives rise to narrow, highly elongated buds in which both actin and septin are tightly polarized to bud tips. Inhibition of Cla4p does not prevent polarization of F-actin, and cytokinesis is blocked only in cells that have not formed a bud before inhibitor treatment; cell polarization and bud emergence are not affected by Cla4p inhibition. Although localization of septin to bud necks is restored in swe1Delta cells, cytokinesis remains defective. Inhibition of Cla4p activity in swe1Delta cells causes a delay of bud emergence after cell polarization, indicating that this checkpoint may mediate an adaptive response that is capable of promoting budding when Cla4p function is reduced. Our data indicate that CLA4 PAK activity is required at an early stage of budding, after actin polarization and coincident with formation of the septin ring, for early bud morphogenesis and assembly of a cytokinesis site. PMID- 11025659 TI - OTRPC4, a nonselective cation channel that confers sensitivity to extracellular osmolarity. AB - Ca2+-permeable channels that are involved in the responses of mammalian cells to changes in extracellular osmolarity have not been characterized at the molecular level. Here we identify a new TRP (transient receptor potential)-like channel protein, OTRPC4, that is expressed at high levels in the kidney, liver and heart. OTRPC4 forms Ca2+-permeable, nonselective cation channels that exhibit spontaneous activity in isotonic media and are rapidly activated by decreases in, and are inhibited by increases in, extracellular osmolarity. Changes in osmolarity of as little as 10% result in significant changes in intracellular Ca2+ concentration. We propose that OTRPC4 is a candidate for a molecular sensor that confers osmosensitivity on mammalian cells. PMID- 11025658 TI - Temporally coordinated assembly and disassembly of replication factories in the absence of DNA synthesis. AB - Here we show that exposure of aphidicolin-arrested Chinese hamster ovary (CHO) cells to the protein-kinase inhibitors 2-aminopurine or caffeine results in initiation of replication at successively later-replicating chromosomal domains, loss of the capacity to synthesize DNA at earlier-replicating sites, release of Mcm2 proteins from chromatin, and redistribution of PCNA and RPA from early- to late-replicating domains in the absence of detectable elongation of replication forks. These results provide evidence that, under conditions of replicational stress, checkpoint controls not only prevent further initiation but may also be required to actively maintain the integrity of stalled replication complexes. PMID- 11025660 TI - New neuropeptides containing carboxy-terminal RFamide and their receptor in mammals. AB - Only a few RFamide peptides have been identified in mammals, although they have been abundantly found in invertebrates. Here we report the identification of a human gene that encodes at least three RFamide-related peptides, hRFRP-1-3. Cells transfected with a seven-transmembrane-domain receptor, OT7T022, specifically respond to synthetic hRFRP-1 and hRFRP-3 but not to hRFRP-2. RFRP and OT7T022 mRNAs are expressed in particular regions of the rat hypothalamus, and intracerebroventricular administration of hRFRP-1 increases prolactin secretion in rats. Our results indicate that a variety of RFamide-related peptides may exist and function in mammals. PMID- 11025661 TI - p53 is associated with cellular microtubules and is transported to the nucleus by dynein. AB - Here we show that p53 protein is physically associated with tubulin in vivo and in vitro, and that it localizes to cellular microtubules. Treatment with vincristine or paclitaxel before DNA-damage or before leptomycin B treatment reduces nuclear accumulation of p53 and expression of mdm2 and p21. Overexpression of dynamitin or microinjection of anti-dynein antibody before DNA damage abrogates nuclear accumulation of p53. Our results indicate that transport of p53 along microtubules is dynein-dependent. The first 25 amino acids of p53 contain the residues that are essential for binding to microtubules. We propose that functional microtubules and the dynein motor protein participate in transport of p53 and facilitate its accumulation in the nucleus after DNA damage. PMID- 11025662 TI - Force production by single kinesin motors. AB - Motor proteins such as kinesin, myosin and polymerase convert chemical energy into work through a cycle that involves nucleotide hydrolysis. Kinetic rates in the cycle that depend upon load identify transitions at which structural changes, such as power strokes or diffusive motions, are likely to occur. Here we show, by modelling data obtained with a molecular force clamp, that kinesin mechanochemistry can be characterized by a mechanism in which a load-dependent isomerization follows ATP binding. This model quantitatively accounts for velocity data over a wide range of loads and ATP levels, and indicates that movement may be accomplished through two sequential 4-nm substeps. Similar considerations account for kinesin processivity, which is found to obey a load dependent Michaelis-Menten relationship. PMID- 11025663 TI - Working strokes by single molecules of the kinesin-related microtubule motor ncd. AB - The ncd protein is a dimeric, ATP-powered motor that belongs to the kinesin family of microtubule motor proteins. Here we resolve single mechanochemical cycles of recombinant, dimeric, full-length ncd, using optical-tweezers-based instrumentation and a three-bead, suspended-microtubule assay. Under conditions of limiting ATP, isolated and transient microtubule-binding events exhibit exponentially distributed and ATP-concentration-dependent lifetimes. These events do not involve consecutive steps along the microtubule, quantitatively confirming that ncd is non-processive. At low loads, a single motor molecule produces ATP triggered working strokes of about 9 nm, which occur at the ends of binding events. PMID- 11025664 TI - The function of PML in p53-dependent apoptosis. AB - The PML gene of acute promyelocytic leukaemia (APL) encodes a growth- and tumour suppresor protein that is essential for several apoptotic signals. The mechanisms by which PML exerts its pro-apoptotic function are still unknown. Here we show that PML acts as a transcriptional co-activator with p53. PML physically interacts with p53 both in vitro and in vivo and co-localizes with p53 in the PML nuclear body (PML-NB). The co-activatory role of PML depends on its ability to localize in the PML-NB. p53-dependent, DNA-damage-induced apoptosis, transcriptional activation by p53, the DNA-binding ability of p53, and the induction of p53 target genes such as Bax and p21 upon gamma-irradiation are all impaired in PML-/- primary cells. These results define a new PML-dependent, p53 regulatory pathway for apoptosis and shed new light on the function of PML in tumour suppression. PMID- 11025665 TI - Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. AB - During carcinogenesis of pancreatic islets in transgenic mice, an angiogenic switch activates the quiescent vasculature. Paradoxically, vascular endothelial growth factor (VEGF) and its receptors are expressed constitutively. Nevertheless, a synthetic inhibitor (SU5416) of VEGF signalling impairs angiogenic switching and tumour growth. Two metalloproteinases, MMP-2/gelatinase A and MMP-9/gelatinase-B, are upregulated in angiogenic lesions. MMP-9 can render normal islets angiogenic, releasing VEGF. MMP inhibitors reduce angiogenic switching, and tumour number and growth, as does genetic ablation of MMP-9. Absence of MMP-2 does not impair induction of angiogenesis, but retards tumour growth, whereas lack of urokinase has no effect. Our results show that MMP-9 is a component of the angiogenic switch. PMID- 11025666 TI - Schnurri mediates Dpp-dependent repression of brinker transcription. AB - Signalling by Decapentaplegic (Dpp), a member of the TGFbeta superfamily of signalling molecules, controls many aspects of Drosophila development by activating and repressing target genes. Several essential components of the Dpp signalling pathway have been identified, including the Dpp receptors Punt and Thick veins (Tkv) as well as the cytoplasmic mediators Mad and Medea. For target genes to be activated, Dpp signalling must suppress transcription of a repressor encoded by the brinker (brk) gene. Here we show that Schnurri (Shn), a large zinc finger protein, is essential for Dpp-mediated repression of brk transcription; in contrast, Shn is not required for target-gene activation. Thus, the Dpp signalling pathway bifurcates, downstream of the signal-mediating SMAD proteins, into a Shn-dependent pathway leading to brk repression and a Shn-independent pathway leading to gene activation. The existence of several Shn-like proteins in vertebrates and the observation that Brk functions in BMP signalling in Xenopus indicates that a similar regulatory cascade may be conserved in higher organisms. PMID- 11025667 TI - A new class of microtubule-associated proteins in plants. AB - In plants there are three microtubule arrays involved in cellular morphogenesis that have no equivalent in animal cells. In animals, microtubules are decorated by another class of proteins - the structural MAPS - which serve to stabilize microtubules and assist in their organization. The best-studied members of this class in plants are the MAP-65 proteins that can be purified together with plant microtubules after several cycles of polymerization and depolymerization. Here we identify three similar MAP-65 complementary DNAs representing a small gene family named NtMAP65-1, which encode a new set of proteins, collectively called NtMAP65 1. We show that NtMAP65-1 protein localizes to areas of overlapping microtubules, indicating that it may function in the behaviour of antiparallel microtubules in the mitotic spindle and the cytokinetic phragmoplast. PMID- 11025668 TI - Cardiolipin provides specificity for targeting of tBid to mitochondria. AB - Recent evidence supports the theory that mitochondrial homeostasis is the key regulatory step in apoptosis through the actions of members of the Bcl-2 family. Pro-apoptotic members of the family, such as Bax, Bad and Bid, can induce the loss of outer-membrane integrity with subsequent redistribution of pro-apoptotic proteins such as cytochrome c that are normally located in the intermembrane spaces of mitochondria. The anti-apoptotic members of the family, such as Bcl-2 and Bcl-XL, protect the integrity of the mitochondrion and prevent the release of death-inducing factors. Bid normally exists in an inactive state in the cytosol, but after cleavage by caspase 8, the carboxy-terminal portion (tBid) moves from cytosol to mitochondria, where it induces release of cytochrome c. Here we address the question of what mediates specific targeting of tBid to the mitochondria. We provide evidence that cardiolipin, which is present in mitochondrial membranes, mediates the targeting of tBid to mitochondria through a previously unknown three-helix domain in tBid. These findings implicate cardiolipin in the pathway for cytochrome c release. PMID- 11025669 TI - Mammalian recombination-repair genes XRCC2 and XRCC3 promote correct chromosome segregation. AB - Growth and development are dependent on the faithful duplication of cells. Duplication requires accurate genome replication, the repair of any DNA damage, and the precise segregation of chromosomes at mitosis; molecular checkpoints ensure the proper progression and fidelity of each stage. Loss of any of these highly conserved functions may result in genetic instability and proneness to cancer. Here we show that highly significant increases in chromosome missegregation occur in cell lines lacking the RAD51-like genes XRCC2 and XRCC3. This increased missegregation is associated with fragmentation of the centrosome, a component of the mitotic spindle, and not with loss of the spindle checkpoint. Our results show that unresolved DNA damage triggers this instability, and that XRCC2 and XRCC3 are potential tumour-suppressor genes in mammals. PMID- 11025670 TI - Threonine 68 is required for radiation-induced phosphorylation and activation of Cds1. AB - In response to DNA damage, eukaryotic cells use a system of checkpoint controls to delay cell-cycle progression. Checkpoint delays provide time for repair of damaged DNA before its replication in S phase and before segregation of chromatids in M phase. The Cds1 (Chk2) tumour-suppressor protein has been implicated in certain checkpoint responses in mammalian cells. It directly phosphorylates and inactivates the mitosis-inducing phosphatase Cdc25 in vitro and is required to maintain the G2 arrest that is observed in response to gamma irradiation. Cds1 also directly phosphorylates p53 in vitro at a site that is implicated in its stabilization, and is required for stabilization of p53 and induction of p53-dependent transcripts in vivo upon gamma-ionizing radiation. Thus, Cds1 functions in both the G1 and G2 checkpoint responses. Like Cds1, the checkpoint protein kinase ATM (ataxia-telangiectasia-mutated) is required for correct operation of both the G1 and G2 damage checkpoints. ATM is necessary for phosphorylation and activation of Cds1 in vivo and can phosphorylate Cds1 in vitro, although evidence that the sites that are phosphorylated by ATM are required for activation is lacking. Here we show that threonine 68 of Cds1 is the preferred site of phosphorylation by ATM in vitro, and is the principal irradiation-induced site of phosphorylation in vivo. The importance of this phosphorylation site is demonstrated by the failure of a mutant, non phosphorylatable form of Cds1 to be fully activated, and by its reduced ability to induce G1 arrest in response to ionising radiation. PMID- 11025671 TI - Award for the ubiquitin pathway. PMID- 11025672 TI - Response PMID- 11025673 TI - Apoptosis and the laws of thermodynamics. PMID- 11025674 TI - Transcriptional regulation: RUPture in the ER. PMID- 11025675 TI - Armadillo takes the APC shuttle. PMID- 11025676 TI - The ins and outs of p53. PMID- 11025677 TI - Chemical genetics: new tools for understanding signalling networks. PMID- 11025678 TI - The crossroads between cell-cell adhesion and motility. PMID- 11025679 TI - Xkid-ding around. PMID- 11025680 TI - Signal transduction takes centre stage. PMID- 11025681 TI - Life is in the bag, or is it? PMID- 11025682 TI - Phosphoinositide biology - messages from lipids PMID- 11025683 TI - Function of Rho family proteins in actin dynamics during phagocytosis and engulfment. AB - Phagocytosis is the uptake of large particles by cells by a mechanism that is based on local rearrangement of the actin microfilament cytoskeleton. In higher organisms, phagocytic cells are essential for host defence against invading pathogens, and phagocytosis contributes to inflammation and the immune response. In addition, engulfment, defined as the phagocytic clearance of cell corpses generated by programmed cell death or apoptosis, has an essential role in tissue homeostasis. Although morphologically distinct phagocytic events can be observed depending on the type of surface receptor engaged, work over the past two years has revealed the essential underlying role of Rho family proteins and their downstream effectors in controlling actin dynamics during phagocytosis. PMID- 11025685 TI - Joseph Parrish, tic doloureux of the bladder and interstitial cystitis. PMID- 11025684 TI - Neoadjuvant hormonal ablative therapy before radical prostatectomy: a review. Is it indicated? AB - PURPOSE: Neoadjuvant hormonal ablation therapy has been used to decrease the rate of positive surgical margins in patients treated with radical prostatectomy. We reviewed the available literature to determine whether this therapy is indicated and beneficial. MATERIALS AND METHODS: We performed a MEDLINE key word search and assessed randomized prospective articles. Data were analyzed for the rate of positive surgical margins, seminal vesicle invasion and lymph node metastasis as well as surgical characteristics, including operative time, blood loss, hospital stay, rate of complications and difficulty of surgical dissection. In addition, these data were evaluated for prostate specific antigen-free survival. RESULTS: Neoadjuvant hormonal therapy decreased the rate of positive margins in 6 of the 7 randomized prospective studies. In none of 4 randomized prospective series was there an improved rate of seminal vesicle invasion with neoadjuvant hormonal therapy. Of 4 studies 3 showed no improvement in the rate of lymph node metastasis after neoadjuvant hormonal therapy compared with that in controls. Similarly there was no improvement in prostate specific antigen-free survival and no significant difference in operative time, operative blood loss, transfusion or hospital stay in patients treated with neoadjuvant hormonal therapy and controls. In addition, in 2 of 3 studies there was no difference in the complication rate. CONCLUSIONS: Analysis of the available literature revealed no significant improvement in outcome to support the routine administration of neoadjuvant hormonal therapy before prostatectomy. PMID- 11025686 TI - Long-term results of sacral nerve stimulation (S3) for the treatment of neurogenic refractory urge incontinence related to detrusor hyperreflexia. AB - PURPOSE: We assess clinical and urodynamic results of sacral nerve stimulation for patients with neurogenic (spinal cord diseases) urge incontinence and detrusor hyperreflexia resistant to parasympatholytic drugs. MATERIALS AND METHODS: Since 1992, 9 women with a mean age of 42.6 years (range 26 to 53) were treated for refractory neurogenic urge incontinence with sacral nerve stimulation. Neurological spinal diseases included viral and vascular myelitis in 1 patient each, multiple sclerosis in 5 and traumatic spinal cord injury in 2. Mean time since neurological diagnosis was 12 years. All patients had incontinence with chronic pad use related to detrusor hyperreflexia. Intermittent self-catheterization for external detrusor-sphincter dyssynergia was used by 5 patients. Social life was impaired and these patients were candidates for bladder augmentation. A sacral (S3) lead was surgically implanted and connected to a subcutaneous neurostimulator after a positive test stimulation trial. RESULTS: Mean followup was 43.6 months (range 7 to 72). All patients had clinically significant improvement of incontinence, and 5 were completely dry. Average number of voids per day decreased from 16.1 to 8.2. Urodynamic parameters at 6 months after implant improved significantly from baseline, including maximum bladder capacity from 244 to 377 ml. and volume at first uninhibited contraction from 214 to 340 ml. Maximum detrusor pressure at first uninhibited contraction increased in 3, stabilized in 2 and decreased in 4 patients. Urodynamic results returned to baseline when stimulation was inactivated. All patients subjectively reported improved visual analog scale results by at least 75% at last followup. CONCLUSIONS: Sacral nerve stimulation can be used as a reversible treatment option for refractory urge incontinence related to detrusor hyperreflexia in select patients with spinal lesions. PMID- 11025687 TI - Clinical use of cystine supersaturation measurements. AB - PURPOSE: We measured the concentration and solubility of cystine in urine from patients with cystinuria or calcium stones and from normal subjects to determine whether urine cystine supersaturation can be calculated from a standard nomogram of solubility versus pH or needs to be measured directly. We also evaluated whether increasing pH of the 24-hour collection recovered enough crystallized cystine to increase cystine supersaturation. MATERIALS AND METHODS: Cystine concentration, pH and usual stone risk factors were measured on 50 ml. aliquots of 24-hour collections from 24 patients with cystinuria, 22 calcium stone formers and 15 normal subjects. After 48 hours of incubation with sodium bicarbonate, a second aliquot was taken from the 24-hour collection for cystine concentration. The original urine at its ambient pH was incubated with an excess of cystine crystals for 24, 48, 72 or 96 hours at 37C to determine solubility and kinetics of equilibration. RESULTS: Cystine solubility varied so widely at any pH range that no predictive nomogram could be relied on for calculating supersaturation. Addition of sodium bicarbonate to the 24-hour urine significantly increased cystine concentration. Urine from stone formers had higher cystine solubility than urine from normal subjects. CONCLUSIONS: Clinical management of cystinuria can be improved by direct measurement of cystine solubility because it varies widely at any given pH. Increasing 24-hour collection pH with sodium bicarbonate additionally improves accuracy of supersaturation measurement by recovering crystallized cystine. PMID- 11025688 TI - Higher urinary potassium is associated with decreased stone growth after shock wave lithotripsy. AB - PURPOSE: We correlated serum and urinary biochemical parameters with radiological evidence of stone growth after shock wave lithotripsy. MATERIALS AND METHODS: Biochemical parameters in serum and 24-hour urine collections of 359 patients were correlated with stone growth for 2 years after shock wave lithotripsy. Each patient underwent a minimum of 2 radiological studies at 3 and 12 months and plain abdominal x-ray at 24 months. The presence and size of stones were documented by a radiologist in blinded fashion. Stone growth was defined as measurable growth of a preexisting stone or new stone formation. RESULTS: A total of 209 patients remained stone-free or had no existing stone growth, while stone size decreased in 30. Of the remaining 120 patients with stone growth 72 had new growth and 48 had growth of preexisting stones. Urinary excretion of potassium was significantly higher in those without than with stone growth (mean 24-hour urine collection plus or minus standard deviation 62 +/- 27 versus 54 +/- 23 mmol., p = 0.009). The only parameter significantly associated with stone growth was urinary potassium. Linear regression revealed that for each 10 unit increase in urinary potassium there was a corresponding 2 mm. decrease in stone growth (p = 0.013). CONCLUSIONS: Our results indicate that increased urinary potassium excretion correlates with a decreased risk of stone growth up to 2 years after shock wave lithotripsy, implying that a high potassium diet may be beneficial for preventing stone growth. The effect of potassium supplementation on stone formation and growth must be investigated further. PMID- 11025689 TI - Influences on renal function in chronic spinal cord injured patients. AB - PURPOSE: The optimal method of bladder management in the spinal cord injured population remains controversial. We determined the significance of bladder management and other factors on renal function in this population. MATERIALS AND METHODS: We retrospectively reviewed the medical records and upper tract imaging studies of 308 patients with a mean followup of 18.7 years since injury. Renal function was assessed by serum creatinine, creatinine clearance and proteinuria measurement, and by upper tract abnormalities on renal ultrasound and nuclear medicine renal scan. Independent variables evaluated for an influence on renal function included patient age, interval since injury, injury level and completeness, vesicoureteral reflux, history of diabetes mellitus and bladder management method. RESULTS: Mean serum creatinine plus or minus standard deviation in patients on chronic Foley catheterization, clean intermittent catheterization and spontaneous voiding was 1.08 +/- 0.99, 0.84 +/- 0.23 and 0.97 +/- 0.45 mg./dl. (analysis of variance p = 0.05, Student's t test p = 0.10), and mean creatinine clearance was 91.1 +/- 46.5, 113.4 +/- 39.8 and 115 +/- 49 ml. per minute, respectively (analysis of variance and Student's t test p <0.01), respectively. Proteinuria was present in 19 patients (6.2%) in the Foley catheterization, 3 (1%) in the clean intermittent catheterization and 4 (1.3%) in the spontaneous voiding group (chi-square test p <0.01), while there were upper tract abnormalities in 56 (18.2%), 20 (6.5%) and 24 (7.8%) patients, respectively (chi-square test p <0.01). Multiple regression analyses revealed no significant predictors of serum creatinine, although older patient age and Foley catheterization significantly predicted low creatinine clearance. Additional logistic regression analyses showed that Foley catheterization was associated with proteinuria and vesicoureteral reflux was associated with upper tract abnormalities. CONCLUSIONS: While renal function may be preserved by all forms of bladder management, chronic indwelling catheters may contribute to renal deterioration. PMID- 11025690 TI - Laparoscopic live donor nephrectomy: the University of Maryland 3-year experience. AB - PURPOSE: We determined whether laparoscopic living donor nephrectomy decreases the morbidity of renal donation for the donor, while providing a renal allograft of a quality comparable to that of open donor nephrectomy. MATERIALS AND METHODS: In a 3-year period laparoscopic donor nephrectomy was performed via the transperitoneal approach. We evaluated donor and recipient medical records for preoperative donor characteristics, intraoperative parameters and complications, and postoperative recovery and complications. RESULTS: Of the 320 laparoscopic donor nephrectomies performed the left kidney was removed in 97.5%. Intraoperative complications, which developed in 10.4% of cases, tended to occur early in the experience and required conversion to open nephrectomy in 5. Average operative time was 31/2 hours and warm ischemia time was 21/2 minutes. As the series progressed, blood loss as well as laparoscopic port size and number decreased but extraction site size remained constant at 7 cm. Urinary retention, prolonged ileus, thigh numbness and incisional hernia were the most common postoperative complications. Postoperative analgesic requirements were low and average hospitalization was 66 hours. CONCLUSIONS: Laparoscopic donor nephrectomy appears to be safe and decreases morbidity in the renal donor. Allograft function is comparable to that in open nephrectomy series. The availability of laparoscopic harvesting may be increasing the living donor volunteer pool. PMID- 11025691 TI - Laparoscopic retroperitoneal live donor right nephrectomy for purposes of allotransplantation and autotransplantation. AB - PURPOSE: We report the technique of and initial experience with retroperitoneal laparoscopic live donor right nephrectomy for purposes of renal allotransplantation and autotransplantation. MATERIALS AND METHODS: A total of 5 patients underwent retroperitoneoscopic live donor nephrectomy of the right kidney for autotransplantation in 4 and living related renal donation in 1. Indications for autotransplantation included a large proximal ureteral tumor, a long distal ureteral stricture and 2 cases of the loin pain hematuria syndrome. In all cases a 3-port retroperitoneal laparoscopic approach and a pelvic muscle splitting Gibson incision for kidney extraction were used. In patients undergoing autotransplantation the same incision was used for subsequent transplantation. RESULTS: All procedures were successfully accomplished without technical or surgical complications. Total mean operating time was 5.8 hours and average laparoscopic donor nephrectomy time was 3.1 hours. Mean renal warm ischemia time, including endoscopic cross clamping of the renal artery to ex vivo cold perfusion, was 4 minutes. Average blood loss for the entire procedure was 400 cc. Radionuclide scan on postoperative day 1 confirmed good blood flow and function in all transplanted kidneys. Mean analgesic requirement was 58 mg. fentanyl. Mean hospital stay was 4 days (range 2 to 8), and convalescence was completed in 3 to 4 weeks. CONCLUSIONS: In the occasional patient requiring renal autotransplantation live donor nephrectomy can be performed laparoscopically with renal extraction and subsequent transplantation through a single standard extraperitoneal Gibson incision, thus, minimizing the overall operative morbidity. Furthermore, these data demonstrate that live donor nephrectomy of the right kidney can be performed safely using a retroperitoneal approach with an adequate length of the right renal vein obtained for allotransplantation or autotransplantation. PMID- 11025692 TI - Metastatic renal cell carcinoma of bone: indications and technique of surgical intervention. AB - PURPOSE: We describe the efficacy of surgical excision of metastatic renal cell carcinoma of bone for achieving local tumor control, pain control and functional outcome with emphasis on the indications and techniques of surgical intervention as well as oncological outcome. MATERIALS AND METHODS: Between 1980 and 1997 we performed surgery on 45 patients (56 lesions) with metastatic renal cell carcinoma of bone. Indications for surgery were solitary bone metastasis, intractable pain, or impending or present pathological fracture. Surgery involved wide excision in 29 cases, marginal excision with adjunctive liquid nitrogen in 25 and amputation in 2. RESULTS: None of the patients had significant bleeding intraoperatively. Mean hospital stay was 9.8 days, during which there was no flap necrosis, deep wound infection, nerve palsy or thromboembolic complication. Postoperatively pain was significantly relieved in 91% of patients, while 89% achieved a good to excellent functional outcome, and 94% with metastatic lesions of the pelvic girdle and lower extremities were ambulatory. Local recurrence developed in only 4 of the 56 lesions (7.1%), including 3 after marginal resection. Survival was more than 2 years in 22 patients (49%) and more than 3 in 17 (38%). CONCLUSIONS: Surgical excision is safe and reliable for restoring mechanical bone stability, relieving pain and providing good function in most patients with metastatic renal cell carcinoma who meet the criteria for surgical intervention. Relatively prolonged survival in these cases justifies considering surgical intervention when feasible. PMID- 11025693 TI - Retrograde ureteroscopic endopyelotomy using the holmium:YAG laser. AB - PURPOSE: We defined the safety and efficacy of retrograde ureteroscopic endopyelotomy using the holmium:YAG laser. METHODS AND MATERIALS: Between July 1996 and December 1999 a total of 28 renal units in 21 women and 6 men 7 to 75 years old (mean age 43.5) with ureteropelvic junction obstruction were treated at our institution with retrograde ureteroscopic endopyelotomy. Ureteropelvic junction obstruction was bilateral in 1 case, primary in 20 and secondary in 8. Endoluminal ultrasound was done before endopyelotomy in all cases. Patients with renal calculi underwent antegrade percutaneous nephrostolithotomy and traditional cold knife endopyelotomy. Endoluminal ultrasound revealed posterior and lateral crossing vessels in 5 patients, who did not undergo the endoscopic approach. Retrograde endopyelotomy was performed using the holmium:YAG laser in 23 cases and electrode incision with pure cutting current in 5. Postoperatively a ureteral stent remained indwelling for an average of 6 weeks. Thereafter patients were followed with serial ultrasound, excretory urography and renal scan at 3 to 6 month intervals. RESULTS: We evaluated 28 upper urinary tracts, including 19 (67.9%) with high insertion ureteropelvic junction obstruction and 9 with an annular stricture. As directed by ultrasound images, the incision location was posterolateral, posterior, lateral and posteromedial in 16, 5, 4 and 3 cases, respectively. Followup was available in all cases at a mean of 10 months (range 3 to 35). Success, defined as improved drainage on radiographic study and absent clinical symptoms, was achieved in 19 of the 23 patients (83%) treated with the holmium:YAG laser. Repeat laser incision resulted in a successful outcome in 2 of the 4 treatment failures. There were no acute surgical complications. CONCLUSIONS: Retrograde ureteroscopic endopyelotomy with the holmium:YAG laser is safe and minimally invasive therapy for primary and secondary ureteropelvic junction obstruction. Endoluminal ultrasound aids in decision making when retrograde endopyelotomy is done. PMID- 11025694 TI - Laparoscopic radical nephroureterectomy for upper tract transitional cell carcinoma: the Cleveland Clinic experience. AB - PURPOSE: We report our single institutional experience with retroperitoneal laparoscopic radical nephroureterectomy in patients with upper tract transitional cell carcinoma and compare results to those achieved by the open technique. MATERIALS AND METHODS: A total of 77 patients underwent radical nephroureterectomy for pathologically confirmed upper tract transitional cell carcinoma. Of these patients 42 underwent laparoscopic nephroureterectomy from September 1997 through January 2000 and 35 underwent open surgery. All specimens were extracted intact. Of the laparoscopic group the juxtavesical ureter and bladder cuff were excised by our novel transvesical needlescopic technique in 27 and radical nephrectomy was performed retroperitoneoscopically in all 42. Data were compared retrospectively with 35 patients undergoing open radical nephroureterectomy from February 1991 through December 1999. RESULTS: Laparoscopy was superior in regard to surgical time (3.7 versus 4.7 hours, p = 0.003), blood loss (242 versus 696 cc, p <0. 0001), specimen weight (559 versus 388 gm., p = 0.04), resumption of oral intake (1.6 versus 3.2 days, p = 0.0004), narcotic analgesia requirements (26 versus 228 mg., p <0.0001), hospital stay (2.3 versus 6.6 days, p <0.0001), normal activities (4.7 versus 8.2 weeks, p = 0.002) and convalescence (8 versus 14.1 weeks, p = 0.007). Complications occurred in 5 patients (12%) in the laparoscopic group, including open conversions in 2, and in 10 (29%) in the open group (p = 0.07). Followup was shorter in the laparoscopic group (11.1 versus 34.4 months, p <0.0001). The 2 groups were similar in regard to bladder recurrence (23% versus 37%, p = 0.42), local retroperitoneal or port site recurrence (0% versus 0%) and metastatic disease (8.6% versus 13%, p = 1.00). Mortality occurred in 2 patients (6%) in the laparoscopic group and 9 (30%) in the open group. Cancer specific survival (97% versus 87%) and crude survival (97% versus 94%) were similar between both groups (p = 0.59). CONCLUSIONS: In patients with upper tract transitional cell carcinoma who are candidates for radical nephroureterectomy the retroperitoneal laparoscopic approach satisfactorily duplicates established technical principles of traditional open oncological surgery, while significantly decreasing morbidity from this major procedure. Short-term oncological and survival data of the laparoscopic technique are comparable to open surgery. Although long-term followup data are not yet available, it appears that laparoscopic radical nephroureterectomy may supplant open surgery as the standard of care in patients with muscle invasive or high grade upper tract transitional cell carcinoma. PMID- 11025695 TI - Upper tract urothelial neoplasms: incidence and survival during the last 2 decades. AB - PURPOSE: Upper tract urothelial cancer is rare but studies in the 1970s showed that its incidence was increasing. We evaluated national trends of the incidence and survival of upper tract urothelial neoplasms from 1973 to 1996. MATERIAL AND METHODS: We obtained information on upper tract urothelial neoplasms from 1973 to 1996 from the National Cancer Institute Surveillance, Epidemiology and End Results program. To provide a time comparison we evaluated upper tract urothelial cancer from 1973 to 1984 and 1985 to 1996. We also calculated overall disease specific survival stratified by cancer stage, patient race and gender for the study period overall. RESULTS: A total of 9,072 cases of upper tract urothelial cancer were identified in the Surveillance, Epidemiology and End Results program from 1973 to 1996, including 5,379 of the renal pelvis and 3,678 of the ureter. Comparing age adjusted annual incidence rates revealed an increase in ureteral neoplasms from 0.69 to 0.73/100,000 person-years but no change in the incidence of renal pelvic tumors, while the rate of in situ neoplasms increased from 7. 2% to 23.1%. Overall disease specific 5-year survival was significantly different in regard to tumor stage (95.1% in situ, 88. 9% localized, 62.6% regional and 16.5% distant lesions). Disease specific annual mortality was greater in black than in white individuals and in women than in men (7.4% versus 4.9% and 6.1% versus 4.4%, respectively). CONCLUSIONS: There appears to have been a slight increase in the national incidence of ureteral tumors in the last 23 years. Fortunately we also detected a slight improvement in the overall disease specific survival of patients with upper tract neoplasms. PMID- 11025696 TI - Laparoscopic management of urachal cysts in adulthood. AB - PURPOSE: Managing persistent and symptomatic urachal anomalies requires wide surgical excision. Such intervention is recommended to prevent symptom recurrence and complications, most notably malignant degeneration. However, traditional open excision is associated with significant morbidity and prolonged convalescence. We report our experience with the laparoscopic excision of urachal remnants as a less morbid, minimally invasive surgical alternative. MATERIALS AND METHODS: Between October 1993 and December 1999, 4 patients with a mean age of 43.3 years who had a symptomatic urachal cyst underwent laparoscopic radical excision of the urachal remnant. Using 2, 10 mm. and 1 or 2, 5 mm. ports the urachus and medial umbilical ligaments were divided at the umbilicus cephalad to the cyst. The specimen, which included the urachus, cyst and medial umbilical ligaments, was then separated from the bladder dome with or without the bladder cuff and removed intact. We reviewed the perioperative records to assess morbidity, recovery and outcome. RESULTS: All 4 procedures were completed successfully. No intraoperative or postoperative complications were reported at a mean followup of 15 months (range 2 to 24). Mean operative time was 180 minutes (range 150 to 210) and average hospital stay was 2.75 days (range 1 to 4). Pathological evaluation confirmed a benign urachal remnant in each case. All patients resumed normal activity within 2 weeks. CONCLUSIONS: To minimize the morbidity of radical excision the laparoscopic management of benign urachal remnants in adulthood is efficacious and our preferred method of management. PMID- 11025697 TI - Early complications of endoscopic treatment for superficial bladder tumors. AB - PURPOSE: Bladder tumors are the second most common tumors of the genitourinary system. Approximately 80% of patients initially present with a superficial lesion, which is treated with transurethral resection. Although transurethral resection is a standard procedure, it is not morbidity-free. We assessed the early complications of transurethral resection for superficial bladder cancer and analyzed various factors that may contribute to its occurrence. MATERIALS AND METHODS: Between January 1979 and December 1996, 2,821 patients with superficial bladder cancer underwent transurethral resection at our center. We assessed intraoperative and immediate postoperative complications of the initial transurethral resection procedure, and correlated them with tumor characteristics. RESULTS: Of the 2,821 patients in our study 2,461 (87%) were male and 360 (13%) were female. Average age was 65 years (range 16 to 94). Of the 145 complications (5.1%) the most common were bleeding in 78 patients (2.8%) and bladder perforation in 36 (1. 3%). Perforation was extraperitoneal in 30 cases (83%) and intraperitoneal in 6 (17%). Conservative treatment and open surgery were done in 32 (89%) and 4 (11%) patients, respectively. We noted no case of tumor seeding. A repeat procedure was done in 77 patients (2.7%) with bleeding as the leading cause of repeat intervention in 65 (84%). Blood transfusion was required in 96 cases (3.4%). The incidence of complications significantly correlated with the size and number of tumors but there was no association with tumor stage, grade or location. CONCLUSIONS: The most common complication of transurethral resection for superficial bladder cancer is bleeding. Currently bladder perforation should be managed conservatively with a minimum risk of extravesical tumor seeding. Our results imply that tumor size and multiple tumor resection are associated with a higher complication rate. PMID- 11025698 TI - The role and impact of pathology review on stage and grade assessment of stages Ta and T1 bladder tumors: a combined analysis of 5 European Organization for Research and Treatment of Cancer Trials. AB - PURPOSE: Pathological interpretations are largely subject to interpathologist and intrapathologist variation. Differences in tumor stage and grade exist in local and review pathological findings in patients with stage Ta-T1 bladder tumors who are entered in randomized trials of adjuvant treatment after transurethral resection. Because they are diagnosed and treated based on local pathological results, it is important to determine the reliability of local pathological evaluations and the extent to which pathology review may change the treatment decision process. MATERIALS AND METHODS: We assessed local and review pathology results in 1,400 patients treated in 5 European Organization for Research and Treatment of Cancer randomized phase III trials comparing various adjuvant prophylactic treatment strategies for primary or recurrent stage Ta-T1 transitional cell bladder cancer. RESULTS: We noted large variations in T category and grade. Pathology review down staged T category to stage Ta in 53% of cases originally classified as stage T1. There was agreement in only 57% and 50% of stage Ta grade 1 and stage T1 grade 3 cases, of which 10% were reclassified as muscle invasive disease greater than stage T1. While T category and grade have prognostic importance, differences in the prognosis based on local and review pathological studies were slight. CONCLUSIONS: Pathology review is not mandatory in low and intermediate risk cases since it has little impact on the prognosis and treatment decision making. In high risk cases of stage T1 grade 3 disease stage or grade is often changed, so that review remains essential in this subgroup. PMID- 11025699 TI - Phase II study of paclitaxel and cisplatin for advanced urothelial cancer. AB - PURPOSE: We examined the role of paclitaxel and cisplatin as first line therapy for metastatic urothelial cancer. MATERIALS AND METHODS: A total of 34 patients were enrolled in this study, and all were eligible for treatment and assessable for response. Patients received 135 mg./m.2 paclitaxel intravenously for 3 hours followed by 70 mg./m.2 cisplatin for 2 hours every 3 weeks to a maximum of 6 cycles. RESULTS: Of the patients 70% experienced a major response to treatment, which was partial/regression in 38% and complete in 32%. Toxicity was manageable with no episodes of grade 4 leukopenia or thrombocytopenia. Nonhematological toxicities included primarily nausea, anorexia and neuropathy, which rarely were severe. CONCLUSIONS: This regimen of paclitaxel and cisplatin is effective, safe and convenient to administer in an outpatient setting for advanced urothelial cancer. PMID- 11025700 TI - Urethroscopic realignment of ruptured bulbar urethra. AB - PURPOSE: We evaluated the efficiency of early endoscopic realignment as primary therapy for bulbar urethral disruption after straddle injury. MATERIALS AND METHODS: From 1990 to 1999 we treated 16 men who had bulbar urethral disruption with endoscopic realignment. Followup included uroflowmetry and urethroscopy at 39 to 85 months. RESULTS: All 16 cases were successfully treated at a single session without intraoperative or postoperative complications. Only 2 patients required intermittent self-dilation once weekly and all were potent during followup. CONCLUSIONS: The results of this minimally invasive procedure are comparable to those of open surgery. It may be performed on an outpatient basis using only local anesthesia. Our results imply that this cost-effective therapy should be done as the initial step in most patients with bulbar urethral disruption. PMID- 11025701 TI - Elevated prostate specific antigen serum levels after intravesical instillation of bacillus Calmette-Guerin. AB - PURPOSE: We determined whether intravesical bacillus Calmette-Guerin (BCG) instillation is associated with elevated prostate specific antigen (PSA). MATERIALS AND METHODS: We treated 36 consecutive patients with bladder cancer with a 6-week course of BCG, followed by cystoscopy at 6 weeks. Blood samples for PSA determination were obtained before each BCG instillation and at cystoscopy with each patient also serving as a control. PSA elevation was defined as 2-fold the baseline level in at least 2 specimens and any PSA level greater than 4 ng./ml. was considered clinically significant. Digital rectal examination was done to identify firm nodules and prostate size. The prostate was examined histologically by transrectal ultrasound guided biopsy or after radical cystectomy. RESULTS: We observed elevated PSA in 27 men (75%) during BCG treatment, of whom 15 (41.6%) had a clinically significant elevation. Overall average PSA increased from 1.3 ng./ml. before BCG instillation to 3.8 during treatment (range 0.1 to 21.5, p <0.0001). In those with a clinically significant elevation average PSA increased from 2.31 ng./ml. at baseline to 6.97 during treatment (p <0.0001) and returned to 3.86 ng./ml. 3 months after treatment. Palpation demonstrated prostatic findings in 10 patients, including firm nodules in 7, while there was significantly elevated PSA in 5 with firm nodules and 2 with diffuse prostatic enlargement. Histological examination of the prostate in 10 patients was diagnostic for granulomatous prostatitis, nonspecific inflammation and benign prostatic hyperplasia in 3, 3 and 4, respectively, of whom none had prostate cancer. CONCLUSIONS: Intravesical BCG therapy is associated with significantly elevated PSA in up to 40% of cases. This effect is self-limited and PSA reverts to normal in 3 months. Therefore, we suggest that prostate biopsy be withheld in such patients and PSA monitored. PMID- 11025702 TI - Prospective identification of National Institutes of Health category IV prostatitis in men with elevated prostate specific antigen. AB - PURPOSE: Although prostatitis may cause elevated prostate specific antigen (PSA), asymptomatic patients are not routinely screened for this diagnosis before transrectal biopsy is performed to rule out cancer. Many negative biopsies reveal evidence of prostatitis classified as National Institutes of Health (NIH) category IV prostatitis or asymptomatic inflammation. To our knowledge this report represents the initial study of the incidence of NIH category IV prostatitis in men before biopsy and its clinical significance. MATERIALS AND METHODS: From 1996 to 1998 asymptomatic men with elevated PSA levels were evaluated for laboratory signs of prostatitis. Patients with expressed prostatic secretions or post-prostate massage urine (voiding bottle 3 [VB3]) positive for greater than 20 and greater than 10 white blood cells per high power field, respectively, received antibiotics for 4 weeks and were reevaluated after 6 to 8 weeks. Men without these clinical signs promptly underwent biopsy. Those with acute urinary tract infection and PSA greater than 30 ng./ml., without a rectum or who refused biopsy were excluded from study. RESULTS: Of the 187 study patients 122 were evaluable with a mean PSA of 9.35 ng./ml., including 51 (42%) with laboratory signs of prostatitis. After treatment PSA was normal in 22 cases and remained elevated in 29, including 9 in which biopsy revealed cancer. The change or improvement in PSA was significantly greater in men with benign results than in those with prostate cancer (-21.32 versus -1.33%, p = 0.001). In the cohort with negative expressed prostatic secretion and VB3 results transrectal ultrasound guided biopsy was done promptly. Screening decreased the number of biopsies by 18% (22 of 122 cases). The positive predictive value of PSA for detecting biopsy proved cancer improved with screening for prostatitis (45 of 122 cases or 37% versus 36 of 71 or 51%). Long-term followup revealed continued normal or stable PSA in the prostatitis cohort. CONCLUSIONS: Screening for NIH category IV prostatitis should be considered in men with elevated PSA. Although patients may be asymptomatic, anxiety caused by prostate cancer and diagnostic procedures contributes to the clinical significance of this disorder. PMID- 11025703 TI - Does the chronic prostatitis/pelvic pain syndrome differ from nonbacterial prostatitis and prostatodynia? AB - PURPOSE: The new consensus classification considers the chronic prostatitis/pelvic pain syndrome (CPPS) based on presence or absence of leukocytes in the expressed prostatic secretions, post-massage urine or seminal fluid analysis. We compared classification based on evaluation of these 3 specimens to the traditional classification based on expressed prostatic secretion examination alone. MATERIALS AND METHODS: A prospective clinical and laboratory protocol was used to evaluate symptomatic patients who had no evidence of urethritis, acute bacterial prostatitis or chronic bacterial prostatitis. RESULTS: Thorough clinical and microbiological evaluation of 310 patients attending our prostatitis clinic was used to select a population of 140 subjects who provided optimal expressed prostatic secretion, post-massage urine and semen specimens. Inflammation was documented in 111 (26%) of 420 samples, including 39 expressed prostatic secretion samples with 500 or greater leukocytes/mm.3, 32 post-massage urine samples with 1 or greater leukocytes/mm.3 and 40 seminal fluid specimens with 1 or greater million leukocytes/mm.3. Of the 140 subjects 73 (52%) had inflammatory chronic prostatitis/pelvic pain according to the consensus criteria but only 39 (28%) had nonbacterial prostatitis according to traditional expressed prostatic secretion criteria (p <0.001). CONCLUSIONS: The new consensus concept of inflammatory chronic prostatitis/pelvic pain includes almost twice as many patients as the traditional category of nonbacterial prostatitis. PMID- 11025704 TI - Filling and voiding symptoms in the American Urological Association symptom index: the value of their distinction in a Veterans Affairs randomized trial of medical therapy in men with a clinical diagnosis of benign prostatic hyperplasia. AB - PURPOSE: We used data from a large Veterans Affairs trial of medical therapy for men with benign prostatic hyperplasia to evaluate the value of calculating separate filling and voiding subscores of the American Urological Association (AUA) symptom index. MATERIALS AND METHODS: We performed factor analysis to assess the psychometric validity of separating the 7 items of the AUA symptom index into filling and voiding subsets. To assess the clinical usefulness of calculating these subscores we correlated them against baseline measurements of symptom interference as well as urodynamic and anatomical measures of disease severity, and used them for predicting the response to medical therapy. RESULTS: Factor analysis confirmed the psychometric validity of separating the AUA symptom index into a 3-item filling and a 4-item voiding subscale. However, calculating filling and voiding subscores did not result in differential correlations with measures of disease interference or severity. It also did not enable us to predict a better symptomatic or uroflowmetry response to medical therapy. CONCLUSIONS: Calculating separate filling and voiding subscores of the AUA symptom index is psychometrically valid but not clinically useful. PMID- 11025705 TI - Transurethral water-induced thermotherapy for the treatment of benign prostatic hyperplasia: a prospective multicenter clinical trial. AB - PURPOSE: We evaluate the effectiveness and safety of transurethral water-induced thermotherapy for the treatment of lower urinary tract symptoms of benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 125 patients with lower urinary tract symptoms due to BPH were enrolled at 8 study centers. Pretreatment evaluation included determination of International Prostate Symptom Score (I-PSS), peak urinary flow rate and quality of life score. Patients also completed a sexual function questionnaire. Patients were evaluated 3, 6 and 12 months after water-induced thermotherapy. RESULTS: Significant improvements in I PSS, peak urinary flow rate and quality of life score were observed as early as 3 months after water-induced thermotherapy. At 12 months I-PSS had improved by a median of 12.5 (95% confidence interval 11.5 to 13.5) versus baseline, peak urinary flow rate by 6.4 ml. per second (5.6 to 7.5) and quality of life score by 2.5 (2.0 to 2.5). I-PSS, peak urinary flow rate and quality of life score improved by 50% or more at 12 months in 61.5%, 71.3% and 71.6% of patients, respectively. No adverse impact of water-induced thermotherapy on sexual function was noted, and preexisting discomfort during ejaculation and interference in sexual function due to lower urinary tract symptoms were significantly ameliorated after treatment. Serious adverse events were infrequent and manageable. CONCLUSIONS: Water-induced thermotherapy significantly alleviates lower urinary tract symptoms of BPH, increases peak urinary flow rate and enhances patient quality of life. This novel catheter based, minimally invasive treatment is easily administered in the outpatient setting. Water-induced thermotherapy holds promise as a useful and cost-effective option for the clinical management of BPH. PMID- 11025706 TI - Prostatic carcinoma: a nutritional disease? Conflicting data from the Kingdom of Saudi Arabia. AB - PURPOSE: The incidence of prostate cancer in Saudi Arabia has been reported to be low at 1.4 to 2.1/100,000 person-years. We prospectively evaluated the true incidence of this disease and its association with dietary factors. MATERIALS AND METHODS: From 1994 to 1997 inclusive Saudi men older than 50 years treated at our institution for various presenting symptoms and diseases were randomly selected from various departments. They were examined prospectively with digital rectal examination, and total and free prostate specific antigen measurement. Transrectal ultrasound and prostatic biopsy were performed when either test was abnormal. Nutrition questionnaires and detailed interviews with a nutritionist were completed to assess the type of diet, and amount of saturated and polyunsaturated fat consumption of patients with prostatic carcinoma and controls. RESULTS: For the 2,270 Saudi men screened we noted an incidence of 3.1/100,000 person-years. Our nutritional survey revealed that recent fat consumption was greater than 120 gm. per person daily, of which about 40% was from meat and dairy products. Saturated fat comprised about 50% of the total fat intake. There was no difference in the amount of fat in the diet of men with and without prostatic carcinoma. CONCLUSIONS: The incidence of prostatic carcinoma in the Kingdom of Saudi Arabia is low despite a high saturated fat diet in recent years. This finding contradicts most western clinical studies, which indicate a positive association of a high fat diet with prostatic carcinoma. PMID- 11025707 TI - A randomized phase 3 study of intraoperative cavernous nerve stimulation with penile tumescence monitoring to improve nerve sparing during radical prostatectomy. AB - PURPOSE: We determine if mapping of the cavernous nerve during radical prostatectomy using intraoperative cavernous nerve stimulation with tumescence monitoring results in improved erectile potency compared to conventional nerve sparing. MATERIALS AND METHODS: A prospective, randomized, single blinded study was performed on 61 patients at 6 centers. Patients had elected to undergo nerve sparing prostatectomy and had normal preoperative erectile function documented by the Sexual Function Inventory Questionnaire (SFIQ) and RigiScan parallel testing. Patients were randomized between conventional nerve sparing and nerve sparing assisted by the CaverMap Surgical Aid. paragraph sign In all patients neural continuity was assessed immediately after prostate removal by proximal cavernous nerve stimulation. All patients were blinded according to their allocation cohort. RESULTS: At 1 year there was substantial improvement in erectile function in the CaverMap group as measured by RigiScan. This group had a mean of 15. 9 minutes of greater than 60% nocturnal tumescence compared to 2.1 minutes in the conventional nerve sparing group (p <0.024). By SFIQ there was a nonsignificant trend to improved potency in the CaverMap group (71% versus 62%, p = 0.17). Of patients who had bilateral, unilateral and no response to stimulation after resection erectile function assessed by SFIQ recovered in 68%, 27% and 0%, respectively (p = 0.016). CONCLUSIONS: CaverMap assisted prostatectomy led to improved erectile function as assessed by RigiScan testing with no associated adverse events. A response to stimulation immediately after removal of the prostate accurately predicted return of erectile function. PMID- 11025708 TI - Bicalutamide monotherapy compared with castration in patients with nonmetastatic locally advanced prostate cancer: 6.3 years of followup. AB - PURPOSE: Nonsteroidal antiandrogen monotherapy may be a treatment option for some patients with advanced prostate cancer. We report a survival and safety update from an analysis of 2 studies in which patients with nonmetastatic (M0) locally advanced disease were treated with either 150 mg. bicalutamide monotherapy or castration. MATERIALS AND METHODS: Data from 2 open label, multicenter studies of identical design were pooled according to protocol. Patients with stage T3/4 prostate cancer were randomized to receive 150 mg. bicalutamide daily or castration (orchiectomy or 3.6 mg. goserelin acetate every 28 days) in a 2:1 ratio. RESULTS: A total of 480 patients with locally advanced prostate cancer were randomized to treatment. After a median followup of 6.3 years mortality was 56%. There was no statistically significant difference between the 2 groups in overall survival (hazard ratio 1.05, upper 1-sided 95% confidence limit 1.31, p = 0.70) or time to progression (1.20, 1.45, p = 0.11). There were statistically significant benefits in the bicalutamide monotherapy group in the 2 quality of life parameters of sexual interest (p = 0.029) and physical capacity (p = 0.046). The highest incidences of adverse events were the pharmacological side effects of hot flashes in the castration group, and breast pain and gynecomastia in the bicalutamide group. The incidences of other types of adverse events were low. Bicalutamide was well tolerated, with few drug related withdrawals from study, and no new safety issues were identified during this longer followup. CONCLUSIONS: Monotherapy with 150 mg. bicalutamide is an attractive alternative to castration in patients with locally advanced prostate cancer for whom immediate hormone therapy is indicated. PMID- 11025709 TI - Whole mounted radical prostatectomy specimens do not increase detection of adverse pathological features. AB - PURPOSE: The optimal method to process radical prostatectomy specimens to maximize the detection of adverse pathological features is unclear and accurate staging is critical. We compare the ability of whole mounted sections to detect these features compared to partially submitted radical prostatectomy specimens. MATERIALS AND METHODS: A total of 93 consecutive radical prostatectomy specimens were processed as whole mounts. Tissue sections were analyzed and the pathological outcomes measured included Gleason score, surgical margin status, and presence or absence of extraprostatic tumor extension and/or seminal vesicle invasion. The pathological outcomes of the preceding cohort were compared to those of a similar cohort consisting of 554 men whose radical prostatectomy specimens were processed as partially submitted glands. RESULTS: A multivariate logistic regression analysis was performed to determine the effect of the method of tissue processing on the pathological outcomes. When considered alone or adjusted for various preoperative patient characteristics (prostate specific antigen, biopsy Gleason score and clinical stage), there were no significant differences in the ability of whole mounted specimens to detect the various outcomes compared to partially submitted specimens (all p >0.4). CONCLUSIONS: Whole mounted sampling of the radical prostatectomy specimen does not improve detection of adverse pathological features. PMID- 11025710 TI - Predicting pathological stage of localized prostate cancer using volume weighted mean nuclear volume. AB - PURPOSE: With the use of prostate specific antigen (PSA) and transrectal ultrasound guided biopsy of the prostate, increasing numbers of clinically localized prostate cancers have recently been detected. Presently, approximately 60% of men newly diagnosed with prostate cancer are believed to have organ confined disease but at the time of surgery less than 50% are ultimately found to have organ confined disease on final pathological analysis. Prediction of pathological stage before surgery using various prognostic factors is needed to determine whether radical prostatectomy is indicated. We have reported that estimates of volume weighted mean nuclear volume can accurately predict the prognosis of clinically localized prostate cancer treated with androgen ablation. In this study we examine whether estimates of mean nuclear volume can predict the pathological stage of cases treated with radical prostatectomy. MATERIALS AND METHODS: A retrospective, prognostic study of 52 patients with clinically localized prostate cancer (21 cases T1c, 27 T2, 4 T3) diagnosed at Kobe City General Hospital between January 1996 and December 1999 and treated with radical prostatectomy was performed. Unbiased estimates of mean nuclear volume measured from transrectal biopsy specimens were compared with PSA at diagnosis, clinical stage, estimated tumor volume and Gleason score with regard to prediction of pathological stage. RESULTS: Univariate analysis revealed that estimates of mean nuclear volume (p <0.0001), PSA at diagnosis (p = 0.0148) and estimated tumor volume (p = 0.0005) significantly correlated with pathological stage but Gleason score did not (p = 0.2011). In addition, multivariate logistic regression analysis demonstrated that estimates of mean nuclear volume (p = 0. 0073), PSA at diagnosis (p = 0.0277) and estimated tumor volume (p = 0.0197) were significantly independent predictors of pathological stage. CONCLUSIONS: The results of our study suggest that combining PSA and estimated tumor volume with estimates of mean nuclear volume can significantly contribute to the prediction of pathological stage of prostate cancer. We recommend use of these 3 factors to predict pathological stage of prostate cancer before surgery. PMID- 11025711 TI - Validation of Partin tables for predicting pathological stage of clinically localized prostate cancer. AB - PURPOSE: The accurate prediction of pathological stage of prostate cancer using preoperative factors is a critical aspect of treatment. In 1997 Partin et al published tables predicting pathological stage using clinical stage, Gleason score and prostate specific antigen (PSA). We tested the validity of the Partin tables. MATERIALS AND METHODS: From 1990 to 1996 inclusively 5,780 patients underwent bilateral pelvic lymphadenectomy and radical prostatectomy for prostate cancer at the Mayo Clinic. However, only 2,475 of these patients met all inclusion criteria of no preoperative treatment, known biopsy Gleason score, available preoperative PSA done either before biopsy or more than 28 days after biopsy and clinical stage T1, T2 or T3a. Among the 2,475 patients 15 had positive lymph nodes and planned prostatectomy was abandoned. The receiver operating characteristics (ROC) curve area, observed and predicted Partin rates of each pathological stage, and positive and negative predictive values were used to compare the Mayo study to the Partin tables. RESULTS: The distribution of pathological stage was organ confined in 67% of Mayo cases versus 48% in the Partin study, extracapsular without seminal vesicle or node involvement in 18% versus 40%, seminal vesicle involvement without nodes in 9% versus 7% and were positive nodes in 6% versus 5%. Using the predicted probabilities of Partin et al the ROC curve area for predicted node positive disease was 0.84 for Mayo cases compared to an estimated 0. 82 in the Partin series. The ROC curve area for predicting organ confined cancer was 0.76 for the Mayo Clinic compared to an estimated 0.73 for the Partin series. The observed rates of node positive disease were similar to those predicted (Partin) based on clinical stage, PSA and Gleason score. For organ confined disease Mayo rates were consistently higher than those predicted from the Partin series using a cut point of 0.50 or greater. Positive and negative predictive values were 0.83 and 0.49 versus 0.63 and 0.70 for the Mayo Clinic and Partin series. CONCLUSIONS: Our study provides strong evidence that sensitivity and specificity of the Partin tables for external clinical sites are similar to what was reported. PMID- 11025712 TI - Preoperative serum prostate specific antigen does not reflect biochemical failure rates after radical prostatectomy in men with large volume cancers. AB - PURPOSE: We compared pathological findings with prostate specific antigen (PSA) failure rates following radical prostatectomy for large volume cancers (6 cc or greater). MATERIALS AND METHODS: A total of 191 men whose radical prostatectomy specimen had a cancer volume of 6 cc or greater were followed for a mean of 3.6 years (range 0.3 to 11.1) and 112 (58.6%) had PSA failure (PSA 0.07 ng./ml. or greater and increasing). Percent Gleason grade 4/5 (the Stanford modified Gleason scale), cancer volume, seminal vesicle invasion, regional lymph nodes, capsular penetration, positive surgical margin, location of the largest cancer in the peripheral or transition zone, prostate weight, patient age, preoperative PSA and clinical stage were analyzed using univariate and multivariate Cox proportional hazards analyses. RESULTS: In univariate regression analysis percent Gleason grade 4/5, lymph node involvement, cancer volume, cancer location in the peripheral zone, capsular penetration and positive surgical margins were significant predictors of biochemical failure. Seminal vesicle invasion, preoperative serum PSA, patient age, prostate weight and clinical stage were not statistically significant. Forward stepwise, multivariate analysis showed that percent Gleason grade 4/5 (p <0.0001, relative risk ratio 2.498), cancer location in the peripheral zone (p = 0.0097, 1.887), cancer volume (p = 0.0157, 1.691) and lymph node involvement (p = 0.0317, 1. 666) were the only independent predictors of biochemical failure. When 52 men with organ confined, large volume prostate cancer were analyzed separately, univariate and multivariate analyses showed that only cancer location in the peripheral zone (p = 0.0021, relative risk ratio 13.473) and percent Gleason grade 4/5 (p = 0. 0449, 4.111) were independent predictors of failure. CONCLUSIONS: Percent Gleason grade 4/5, cancer location in the peripheral zone, cancer volume and lymph node involvement have prognostic value in large volume prostate cancer. Cancer location in the peripheral zone and percent Gleason grade 4/5 are the most powerful predictors of biochemical failure in men whose cancer is 6 cc or greater and contained in the prostatic capsule. Preoperative serum PSA is not helpful in distinguishing biochemical failure rates in these large volume cancers whether they are organ confined or not. PMID- 11025713 TI - Predicting outcomes after radical prostatectomy. PMID- 11025714 TI - Use of the holmium:YAG laser for the impacted stone basket. AB - PURPOSE: During ureteroscopic basketing of ureteral calculi a stone may become engaged in the basket and the basket impacted in the ureter. We describe an endoscopic technique of managing the impacted basket and stone. MATERIALS AND METHODS: The ureteroscope is back loaded off of the basket, and passed into the ureter beside the basket and stone. The holmium:YAG laser is used to irradiate a basket wire, fracturing the wire and releasing the stone. The basket is removed and the stone managed by holmium:YAG lithotripsy. RESULTS: We successfully treated 3 consecutive patients with this technique. No injuries were observed. CONCLUSIONS: The holmium:YAG laser may be used to free an impacted stone basket containing a ureteral calculus. PMID- 11025715 TI - A simple technique to change an obstructed ureteral stent in patients with an ileal conduit. AB - PURPOSE: We describe a simple technique to replace an obstructed ureteral stent in patients with an ileal conduit and ureteral stenosis. MATERIALS AND METHODS: Our study included 4 patients with an ileal conduit and ureteral stenosis. A total of 10 obstructed ureteral stents were replaced with a new stent using this simple technique under radiographic guidance. RESULTS: All obstructed ureteral stents were successfully replaced with a new stent without endoscopy, percutaneous nephrostomy or any special devices. CONCLUSIONS: Our technique is simple, easy and highly successful for changing an obstructed ureteral stent. PMID- 11025716 TI - Dynamic half Fourier acquisition, single shot turbo spin-echo magnetic resonance imaging for evaluating the female pelvis. AB - PURPOSE: We assessed the merit of dynamic half Fourier acquisition, single shot turbo spin-echo sequence T2-weighted magnetic resonance imaging (MRI) for evaluating pelvic organ prolapse and all other female pelvic pathology by prospectively correlating clinical with imaging findings. MATERIALS AND METHODS: From September 1997 to April 1998, 100 consecutive women 23 to 88 years old with (65) and without (35) pelvic organ prolapse underwent half Fourier acquisition, single shot turbo spin-echo sequence dynamic pelvic T2-weighted MRI at our institution using a 1.5 Tesla magnet with phased array coils. Mid sagittal and parasagittal views with the patient supine, relaxed and straining were obtained using no pre-examination preparation or instrumentation. We evaluated the anterior vaginal wall, bladder, urethra, posterior vaginal wall, rectum, pelvic floor musculature, perineum, uterus, vaginal cuff, ovaries, ureters and intraperitoneal organs for all pathological conditions, including pelvic prolapse. Patients underwent a prospective physical examination performed by a female urologist, and an experienced radiologist blinded to pre-imaging clinical findings interpreted all studies. Physical examination, MRI and intraoperative findings were statistically correlated. RESULTS: Total image acquisition time was 2.5 minutes, room time 10 minutes and cost American $540. Half Fourier acquisition, single shot turbo spin-echo T2-weighted MRI revealed pathological entities other than pelvic prolapse in 55 cases, including uterine fibroids in 11, ovarian cysts in 9, bilateral ureteronephrosis in 3, nabothian cyst in 7, Bartholin's gland cyst in 4, urethral diverticulum in 3, polytetrafluoroethylene graft abscess in 3, bladder diverticulum in 2, sacral spinal abnormalities in 2, bladder tumor in 1, sigmoid diverticulosis in 1 and other in 9. Intraoperative findings were considered the gold standard against which physical examination and MRI were compared. Using these criteria the sensitivity, specificity and positive predictive value of MRI were 100%, 83% and 97% for cystocele; 100%, 75% and 94% for urethrocele; 100%, 54% and 33% for vaginal vault prolapse; 83%, 100% and 100% for uterine prolapse; 87%, 80% and 91% for enterocele; and 76%, 50% and 96% for rectocele. CONCLUSIONS: Dynamic half Fourier acquisition, single shot turbo spin echo MRI appears to be an important adjunct in the comprehensive evaluation of the female pelvis. Except for rectocele, pelvic floor prolapse is accurately staged and pelvic organ pathology reliably detected. The technique is rapid, noninvasive and cost-effective, and it allows the clinician to visualize the whole pelvis using a single dynamic study that provides superb anatomical detail. PMID- 11025717 TI - Value of routine evaluation of the voiding phase when performing urodynamic testing in women with lower urinary tract symptoms. AB - PURPOSE: Lower urinary tract symptoms in women are often evaluated by cystometrography. We only assessed the bladder response to filling and not the impact of abnormal voiding, which is known to cause lower urinary tract symptoms. We determined the prevalence of voiding abnormalities in women with lower urinary tract symptoms and compared cystometrography to cystometrography plus voiding pressure flow study for evaluating this condition. We also determined whether storage or voiding symptoms predicted abnormal voiding. MATERIALS AND METHODS: We reviewed the records of 134 women who underwent video urodynamics with cystometrography and voiding pressure flow study to evaluate lower urinary tract symptoms. Patients with a history of neurological disease, grade 4 pelvic prolapse or a primary complaint of stress incontinence were excluded from study. All participants completed an American Urological Association symptom index and scores were subclassified as total, storage and voiding. A diagnosis was made in each case based on cystometrography findings, while any additional diagnoses when applicable were based on the voiding pressure flow study. Symptom scores were compared in women in whom the voiding study did and did not add information. RESULTS: Mean patient age was 53.1 years (range 19 to 90). Voiding studies added information in 44 cases (33%), including dysfunctional voiding in 16, obstruction due to a moderate cystocele in 6, primary bladder neck obstruction in 6, external detrusor sphincter dyssynergia as the initial presentation of neurological disease in 5, obstruction after incontinence surgery in 3, urethral stricture in 3, post-void contraction mimicking symptoms in 2, impaired contractility in 2 and an obstructing urethral diverticulum in 1. A total of 32 patients (24%) did not void during the study. Those with voiding abnormalities had higher total and voiding but similar storage symptom scores (23.1 versus 18.5, 12.3 versus 8.0 and 10.8 versus 10.5 points, p = 0.0008, 0.0001 and 0.58, respectively). CONCLUSIONS: Women with lower urinary tract symptoms may have voiding abnormalities that are missed by cystometrography only. Voiding studies are useful for properly diagnosing and treating such cases. Women with abnormal voiding seem to have more severe voiding symptoms than those without such abnormalities. Occult neurological disease may also be identified in patients with lower urinary tract symptoms and voiding abnormalities. PMID- 11025718 TI - Results of transurethral injection of silicone micro-implants for females with intrinsic sphincter deficiency. AB - PURPOSE: We evaluated the medium term efficiency of silicone micro-implants injected in women with intrinsic sphincter deficiency. MATERIALS AND METHODS: A total of 21 women with intrinsic sphincter deficiency underwent transurethral injection of silicone micro-implants between August 1996 and February 1997. Each patient was assessed preoperatively by questionnaire, physical examination and urodynamic study. The results were evaluated by questionnaire at 1 month, and 1 and 2 years after silicone injection. The outcome was classified as dry in all circumstances, improved or failure. RESULTS: All patients (median age 68 years, range 46 to 83) had undergone previous anti-incontinence or prolapse surgeries. At 1 month 2 patients (10%) were dry and 9 (42%) were improved, and treatment failed in 10 (48%). At 1 year (median 16 months, range 14 to 22) 2 cases (10%) were classified as dry, 8 (38%) improved and 11 (52%) failures. At last followup (median 31 months, range 24 to 34) 4 cases (19%) were classified as dry, 6 (29%) improved and 11 (52%) failures. None of the 6 patients with bladder neck hypermobility was dry. CONCLUSIONS: Our results of silicone transurethral injection are disappointing but comparable to other bulking agents without a time dependent decrease in efficiency. The use of silicone micro-implants is an alternative for the treatment of intrinsic sphincter deficiency in patients without bladder neck hypermobility and in whom the sling procedure has failed. PMID- 11025719 TI - Prospective analysis of 373 consecutive women with stress urinary incontinence treated with a vaginal wall sling: the Columbia-Cornell University experience. AB - PURPOSE: We compare the efficacy and morbidity of 373 consecutive women who underwent a vaginal wall sling for stress urinary incontinence due either to anatomical incontinence or intrinsic sphincter deficiency. To our knowledge this series is the largest prospective database on surgical management of stress urinary incontinence in the urological literature. MATERIALS AND METHODS: Preoperative evaluation included history, voiding diary, physical examination, cystoscopy, pad count and video urodynamic study. Outcome measures included postoperative presence of incontinence secondary to either stress and/or detrusor instability, number of pads used, complications, operating time, length of suprapubic catheterization, length of hospitalization and loss of work days. RESULTS: A total of 373 consecutive women 18 to 85 years old (mean age 55.7) were followed for a mean of 39.8 months. Of these patients 183 (49%) presented with anatomical incontinence and the remaining 190 (51%) had intrinsic sphincter deficiency. Preoperative detrusor instability was present in 60 (33%) patients with anatomical incontinence and 68 (36%) with intrinsic sphincter deficiency. Postoperatively, 14 patients (4%) had recurrent stress urinary incontinence. De novo detrusor instability and urge incontinence were noted in 30 women (8%), and was persistent in 22 (6%). There was no correlation between the diagnosis of anatomical incontinence or intrinsic sphincter deficiency and persistent stress urinary incontinence or detrusor instability. Daily pad use was decreased from 4.3 to 0.5 and from 4.6 to 0.4, respectively, for patients with anatomical incontinence and intrinsic sphincter deficiency. Operating time, catheter duration, length of hospital stay and days lost from work for patients with anatomical incontinence (33.3 +/- 14.3 minutes, 4.7 +/- 1.1 days, 0.9 +/- 0.7 days and 11.3 +/- 2.9 days, respectively) were similar to patients with intrinsic sphincter deficiency (38.4 +/- 17.8, 4.6 +/- 0.9, 1.1 +/- 0.7, 12.4 +/- 4.7). The most common complications were urinary tract infection (3%), wound infection (4%) and pelvic organ prolapse (7%). CONCLUSIONS: The results of this large database suggest that the vaginal wall sling is effective for the management of stress urinary incontinence. Efficacy, morbidity and reduced hospitalization time were similar for patients with either anatomical incontinence or intrinsic sphincter deficiency and independent of surgeon experience. PMID- 11025720 TI - Resolution of urge symptoms following sling cystourethropexy. AB - PURPOSE: Persistence of urinary urge symptoms in women after pubovaginal sling cystourethropexy is a distressing problem. We assess the ability of preoperative video urodynamics to identify patients in whom urinary urge symptoms failed to resolve postoperatively. MATERIALS AND METHODS: The records of 84 consecutive women who had undergone pubovaginal sling cystourethropexy were reviewed. Preoperative video urodynamics revealed stress urinary incontinence in all cases and concomitant detrusor instability in some. Cases of motor urge, defined as urge symptoms plus detrusor instability, were divided into low and high pressure categories. Patients with urge symptoms but no demonstrable detrusor instability on video urodynamics were diagnosed with sensory urge. Postoperatively complete urge resolution was defined as total absence of symptoms without pharmacological therapy. Mean followup was 26.7 months (range 2 to 62). RESULTS: Of the 84 patients 41 with motor urge and 28 with sensory urge had preoperative urge symptoms. Complete resolution or improvement in urge symptoms occurred in 24 (58.5%) and 7 (17.1%) of the motor urge, and 11 (39.3%) and 9 (32.1%) of sensory urge cases, respectively. Of the 41 patients with motor urge 23 had low pressure instability, which completely resolved in 21 (91.3%) and improved in 2 (8.7%). Of the 18 remaining patients with high pressure motor urge only 5 (27. 8%) had complete resolution of urge and 5 (27.8%) had improvement. CONCLUSIONS: Our results suggest that patients with low pressure motor urge are significantly more likely to experience resolution of urinary urge symptoms after pubovaginal sling cystourethropexy than those with high pressure motor urge or sensory urge. PMID- 11025721 TI - Injections, slings and urodynamics. PMID- 11025722 TI - Cadaveric versus autologous fascia lata for the pubovaginal sling: surgical outcome and patient satisfaction. AB - PURPOSE: We report our initial experience with cadaveric fascia lata in pubovaginal sling procedures. MATERIALS AND METHODS: We compared 121 consecutive women who underwent a sling procedure using cadaveric fascia lata from February 1997 through June 1999 (group 1) with 46 consecutive women who underwent a sling procedure using autologous fascia lata from May 1994 through July 1997 (group 2). RESULTS: Mean followup was longer in group 2 (44 versus 12 months). A total of 104 of the 121 group 1 patients (86%) responded to the questionnaire, of whom 85% were cured of stress incontinence, 83% reported overall improvement in urinary control and 74% had no or minimal leakage not requiring pads. Median catheterization time was 9 days (range 4 to 120). Overall 89% of the women were satisfied with the results and 83% would recommend this surgery. A total of 30 of the 46 group 2 patients (65%) responded to the questionnaire, of whom 90% were cured of stress incontinence, 90% reported overall improvement in urinary control and 73% had no or minimal leakage not requiring pads. Median catheterization time was 14 days (range 6 to 180). Overall 90% of the women were satisfied with the results and 83% would recommend this surgery. CONCLUSIONS: Cadaveric fascia lata pubovaginal slings appear to be safe. Early experience suggests that cadaveric fascia lata may be considered an alternative to autologous fascia. Cadaveric and autologous fascia lata appear to have a high success rate. PMID- 11025723 TI - New onset vesicovaginal fistula after transurethral collagen injection in women who underwent cystectomy and orthotopic neobladder creation: presentation and definitive treatment. AB - PURPOSE: We present our experience with collagen injection for treating urinary incontinence after cystectomy and orthotopic bladder substitution in women. We discuss the efficacy of collagen injection, specific complications and subsequent definitive therapy. MATERIALS AND METHODS: We performed cystectomy and orthotopic bladder substitution in 2 women for muscle invasive transitional cell carcinoma of the bladder. In each case new onset stress urinary incontinence developed after surgery that was refractory to conservative therapy. Intrinsic sphincter deficiency was diagnosed in each patient by video urodynamic studies. Initial treatment involved transurethral collagen injections but subsequent intervention was required due to resultant complications and primary therapy inefficacy. RESULTS: Collagen (3.5 cc per session) was injected in 1 case at 2 treatment sessions and in the other at 3. Incontinence symptoms did not significantly improve in either patient and a new onset vesicovaginal fistula developed 2 days and 1 month after collagen injection, respectively. Subsequently in each case 1 stage transvaginal primary fistula repair was done in multiple layers with a pubovaginal sling procedure. Six months after repair there has been no recurrent fistula and the women remain hypercontinent, requiring intermittent self catheterization. They are satisfied with their eventual lower tract function and overall outcome. CONCLUSIONS: Collagen injection for type 3 stress urinary incontinence after cystectomy and orthotopic bladder replacement in women may not be as effective and innocuous as in patients with a native bladder. Initial treatment with a pubovaginal sling procedure should be considered. PMID- 11025724 TI - Voiding dysfunction and Parkinson's disease: urodynamic abnormalities and urinary symptoms. AB - PURPOSE: We evaluated bladder dysfunction and Parkinson's disease in regard to disease severity and determined whether subjective patient urinary symptoms correlated with urodynamic abnormalities. MATERIALS AND METHODS: We assessed bladder dysfunction in 70 patients with Parkinson's disease and urinary symptoms using the International Prostate Symptom Score and urodynamic tests. RESULTS: Urodynamic evaluation revealed detrusor hyperreflexia in 47 patients (67%), hyporeflexia or areflexia in 11 (16%), hyperreflexia with impaired contractile function in 6 (9%), hyperreflexia with detrusor-sphincter dyssynergia in 2 (3%) and normal function in 4 (6%). The incidence of urodynamic abnormalities appeared to increase with disease severity. However, the only urodynamic parameter that correlated with disease severity was post-void residual urine volume. On the other hand, symptom index scores increased with disease severity. The irritative symptom score correlated with maximum cystometric capacity and volume at initial desire to void, whereas the obstructive symptom score correlated with post-void residual urine volume. Also, irritative and obstructive scores were good predictors of overactivity during the storage and underactivity at the voiding phases. CONCLUSIONS: Bladder function may deteriorate progressively with advancing disease. Symptom scores are fairly accurate for predicting likely urodynamic abnormalities. Our results imply that quantifying subjective urinary symptoms is useful for estimating the severity and type of bladder dysfunction. PMID- 11025725 TI - Anastomotic calculus formation after side-to-side ureteroureterostomy in a patient with complete ureteral duplication. PMID- 11025727 TI - Intracavernous penile hematoma. PMID- 11025726 TI - Urethral recurrence of transitional cell carcinoma in a female patient after cystectomy and orthotopic ileal neobladder. PMID- 11025728 TI - Spontaneous thrombosis of the deep dorsal penile vein in a patient with thrombophilia. PMID- 11025729 TI - Penile salvage for squamous cell carcinoma associated with mineral oil injection. PMID- 11025730 TI - A hypoechoic lesion found on testicular ultrasound after testicular piercing. PMID- 11025731 TI - Testicular histoplasmosis. PMID- 11025732 TI - HIV infection presenting as an unusually large pure yolk sac tumor of the testis. PMID- 11025733 TI - Metastatic testis tumor presenting as gastrointestinal hemorrhage. PMID- 11025734 TI - Genital self-mutilation and the Internet. PMID- 11025735 TI - Idiopathic bilateral panniculitis of the spermatic cord in an elderly male patient. PMID- 11025736 TI - Male infertility due to a benign prostatic polyp. PMID- 11025737 TI - Prostate brachytherapy seed migration to the right ventricle found at autopsy following acute cardiac dysrhythmia. PMID- 11025738 TI - Local invasion of prostate carcinoma causing vesicorectal fistula. PMID- 11025739 TI - Trimethoprim-sulfamethoxazole induced recurrent aseptic meningitis. PMID- 11025740 TI - Re: A randomized double-blind trial of oral l-arginine for treatment of interstitial cystitis. PMID- 11025741 TI - Re: Efficacy and safety of valrubicin for the treatment of Bacillus Calmette Guerin refractory carcinoma in situ of the bladder. PMID- 11025742 TI - RE: Cowper's syringocele: symptoms, classification and treatment of an unappreciated problem. PMID- 11025743 TI - Re: Surgical management of urinary incontinence in children with neurogenic sphincteric incompetence. PMID- 11025744 TI - Re: Preoperative treatment with human chorionic gonadotropin in infancy decreases the severity of proximal hypospadias and chordee. PMID- 11025745 TI - Re: Y-to-I wrap: use of the distal spongiosum for hypospadias repair. PMID- 11025746 TI - 99m Technetium dimercapto-succinic acid renal scintigraphy abnormalities in infants with sterile high grade vesicoureteral reflux. AB - PURPOSE: Although vesicoureteral reflux associated with bacteriuria may cause renal scarring, sterile reflux is thought not to cause renal injury. We determined the incidence and associated characteristics of renal abnormalities using 99mtechnetium(Tc) dimercapto-succinic acid (DMSA) renal scintigraphy in infants with high grade vesicoureteral reflux but no history of urinary tract infection. MATERIALS AND METHODS: We retrospectively reviewed the results of 99mTc-DMSA renal scintigraphy and renal ultrasonography performed during the first 6 months of life in infants with vesicoureteral reflux detected during the postnatal evaluation of prenatal hydronephrosis or sibling reflux screening. Those with a history of urinary tract infection, or evidence of ureteropelvic junction or bladder outlet obstruction were excluded from study. RESULTS: Of the 28 male and 6 female infants who met study criteria vesicoureteral reflux was bilateral in 25 and unilateral in 9. Reflux grade was IV or V, II or III and I in 38, 18 and 3 of the 59 refluxing renal units, respectively. 99mTc-DMSA renal scintigraphy revealed parenchymal abnormalities in 24 refluxing renal units (41%) in 22 patients (65%), of whom 19 (86%) were male and 15 (68%) had bilateral reflux. We noted differential uptake less than 40% with and without cortical defects in 10 and 7 refluxing units, respectively, and cortical defects only in 7. Of the 24 refluxing units with abnormalities 21 were associated with grade IV or V and 3 with grade II or III reflux. Ultrasound showed evidence of renal injury in only 7 of the 17 patients (41%) in whom 99mTc-DMSA scintigraphy was abnormal. CONCLUSIONS: In our study the majority of infants with high grade reflux had decreased differential function and/or cortical defects. Parenchymal defects detected by 99mTc-DMSA renal scintigraphy were often not identified by renal ultrasound. Therefore, 99mTc-DMSA renal scintigraphy is especially useful for initially evaluating infants with high grade, sterile vesicoureteral reflux. PMID- 11025747 TI - Injectable small intestinal submucosa: preliminary evaluation for use in endoscopic urological surgery. AB - PURPOSE: We evaluated the possible use of small intestinal submucosa in endoscopic urological surgery by assessing the smooth muscle regenerative capabilities and physical response of various forms of injectable small intestinal submucosa in the canine model. MATERIALS AND METHODS: In blinded fashion we injected small intestinal submucosa in 12 dogs submucosally under direct vision using a 20 gauge endoscopic needle. The 4 small intestinal submucosa formulations varied in harvesting method and sterilization technique. Animals were divided into groups of 3 and sacrificed 2 weeks, 6 weeks, 3 months and 6 months after surgery. Each injection site was analyzed grossly and histologically. Smooth muscle regeneration was identified by alpha-smooth muscle actin immunohistochemical staining. RESULTS: We identified 2 injectable small intestinal submucosa formulations that induced progressive smooth muscle regeneration at the site of submucosal injection compared with controls. De novo smooth muscle cells appeared in single cell aggregates as early as 6 weeks and in globular aggregates at 3 months. By 6 months early muscle bundle formation was noted. These 2 injectable small intestinal submucosa formulations also had the best submucosal volume preservation of about 25% of injected material during the study period. CONCLUSIONS: Injectable small intestinal submucosa promotes progressive submucosal smooth muscle regeneration in the canine bladder. The combined regenerative and bulking abilities of injectable small intestinal submucosa make this compound unique and novel. The clinical usefulness of injectable small intestinal submucosa for endoscopic correction of reflux and incontinence deserves further investigation. PMID- 11025748 TI - Biofeedback training for detrusor overactivity in children. AB - PURPOSE: We evaluated biofeedback training for incontinence due to detrusor overactivity in children. MATERIALS AND METHODS: Included in our study were 22 boys and 17 girls with a mean age of 11.2 years. We noted nighttime incontinence in 3 patients, nighttime incontinence and daytime urinary symptoms in 26, and daytime incontinence in 10. All patients had detrusor overactivity and incontinence refractory to conventional treatment, including bladder training, tricyclic antidepressants, anticholinergics, desmopressin and/or conditioning therapy. Urodynamic study was performed using an 8Fr double lumen transurethral catheter for cystometry, a double balloon transrectal catheter for rectal pressure and external anal sphincter pressure measurement, and surface electrodes for sphincter electromyography. During biofeedback training patients were instructed to contract the anal sphincter without raising abdominal pressure to inhibit overactive bladder contractions. Biofeedback training was repeated monthly until cystometry revealed a stable bladder or lower urinary tract symptoms improved considerably. RESULTS: Four patients were lost to followup. Of the remaining 35 children urinary symptoms were cured in 23 and improved in 4. Urodynamic studies after 6 months of biofeedback training in 33 cases showed that bladder overactivity disappeared in 10 and improved in 18. Bladder capacity at the initial desire to void and maximum cystometric capacity increased significantly (p = 0.0115 and <0.0001, respectively). Detrusor-sphincter dyssynergia in 2 patients before biofeedback training resolved in each after therapy. CONCLUSIONS: Biofeedback training for detrusor overactivity is effective even in pediatric cases refractory to conventional treatment. PMID- 11025749 TI - The use of free grafts for correcting penile chordee. AB - PURPOSE: Patients with extensive penile chordee may require free graft placement to create a straight phallus. We review our experience with those who underwent primary (no previous surgery) and secondary repair using dermal and tunica vaginalis grafts to correct penile chordee. MATERIALS AND METHODS: Between 1992 and 1998, 28 patients required free graft placement to correct penile chordee. In 8 cases (29%) primary repair was done using tunica vaginalis in 2 and a dermal graft in 6, while in 20 (71%) secondary repair was performed due to recurrent chordee with or without hypospadias. Tunica vaginalis was used in 3 patients and dermal grafts in 17. Grafts were used when straightening did not result from penile shaft degloving, dorsal plication and urethral plate division. Mean followup was 2 years. RESULTS: Residual chordee developed in 1 of 8 patients after primary and in 2 of 20 after secondary repair. Tunica vaginalis grafts had been placed at repair in all patients with residual chordee but there was no recurrent chordee after dermal graft placement. Chordee recurred in 60% of the patients who received a tunica vaginalis graft. CONCLUSIONS: Conventional techniques, such as penile degloving and dorsal plication, may be used to correct penile chordee in most cases, while free grafts are more likely to be required for secondary repair. We believe that dermal grafts result in more successful repair than tunica vaginalis grafts, which seem to be associated with a higher incidence of residual chordee. PMID- 11025750 TI - Elevated placental estradiol: a possible etiological factor of human cryptorchidism. AB - PURPOSE: It has been convincingly argued that the increasing incidence of reproductive abnormalities in human males may be associated with increased estrogen exposure during gestation. We documented the expression of estradiol in the syncytiotrophoblast and placenta of males born with cryptorchidism and normal genitalia, respectively. MATERIALS AND METHODS: Tissue from newborn placentas was fixed in glutaraldehyde and embedded in ePON for immunohistological procedures. Cryptorchidism was unilateral in 6 males and bilateral in 1. Semithin histological sections of placental biopsies of these 7 males and 7 randomly selected, normal male placentas were analyzed immunohistochemically with a polyclonal anti-estradiol-17 beta, 6-keto antibody. RESULTS: The weak expression of estradiol in the placentas of normal males was localized predominantly at the basal part of the syncytiotrophoblast in the terminal placental villi. In contrast, all placentas of cryptorchid males had strong expression of estradiol at the basal portion of the syncytiotrophoblast. CONCLUSIONS: The increased expression of estradiol in the syncytiotrophoblast may have an impact on testicular descent. PMID- 11025751 TI - Excess fetal estrogen as an etiological cause of human cryptorchidism. PMID- 11025752 TI - Paternity and hormone levels after unilateral cryptorchidism: association with pretreatment testicular location. AB - PURPOSE: We determined differences in paternity and levels of the hormones inhibin B, follicle- stimulating hormone, luteinizing hormone, testosterone and free testosterone based on the preoperative location of the undescended testis in men with previous unilateral cryptorchidism. MATERIALS AND METHODS: Testicular location was determined by a review of the medical records and paternity or attempted paternity using a detailed questionnaire administered to 320 men with previous unilateral cryptorchidism. In 103 cases we performed semen analysis and measured the levels of the hormones inhibin B, luteinizing hormone, follicle stimulating hormone, testosterone and free testosterone. Paternity, sperm count and hormonal parameters were compared with cryptorchid testicular location using analysis of variance and chi-square analysis. Logistic regression was done to analyze pretreatment testicular location as a risk factor for infertility. RESULTS: Paternity, duration of attempted conception in men who achieved paternity, sperm count and hormone levels did not differ based on pretreatment abdominal, internal ring, inguinal canal, external ring, upper scrotum or ectopic testicular location. The overall paternity rate was 90% with the lowest rate of 83.3% in the abdominal group. More than 12 months were required to achieve conception in 28.9% of the study group overall and in 39.4% of the abdominal group. Varicocele and a partner with fertility problems were risk factors for infertility, while abdominal testicular location caused borderline significant risk. CONCLUSIONS: Preoperative testicular location in men with previous unilateral cryptorchidism is not a major determinant of fertility according to paternity, sperm count or hormone levels. PMID- 11025753 TI - The anatomical approach to inguinal orchiopexy. AB - PURPOSE: Achieving a scrotally positioned testis with no iatrogenic atrophy is the primary goal of the surgical treatment of cryptorchidism. The success rate is associated with preoperative testicular position and the technique used. Our success rate with inguinal orchiopexy for abdominal testis is higher than those previously published and may be due to our anatomical approach to retroperitoneal dissection. We review the relevant anatomy and discuss the surgical technique of inguinal orchiopexy. MATERIALS AND METHODS: We researched and reviewed the relevant technical and anatomical literature on inguinal orchiopexy, and describe the anatomical approach to orchiopexy in detail. RESULTS: An understanding of the retroperitoneal fascial layers is the key to understanding surgery of the retroperitoneum. Dividing the internal spermatic fascia is required for the surgeon to separate the processus vaginalis from the vas and vessels. CONCLUSIONS: The average age of children undergoing cryptorchid surgery has decreased dramatically during the last 40 years. In such young patients extensive mobilization of the vas and vessels may be accomplished through a relatively small incision. We believe that our success rate with inguinal orchiopexy for abdominal testis is associated with our anatomical approach to retroperitoneal dissection. PMID- 11025754 TI - Inguinal reoperation for undescended testis and hernia: approach to the spermatic cord through the cremaster fascia. AB - PURPOSE: Reoperation of the inguinal canal is difficult with few available lucid descriptions of the technique. We reviewed our experience with a unique surgical approach to the spermatic cord through the cremaster fascia in patients who had undergone previous operations for undescended testis or hernia and report its outcome. MATERIALS AND METHODS: A total of 54 inguinal reoperations were performed in 45 boys with an undescended testis or indirect inguinal hernia using a unique surgical approach through the cremaster fascia. RESULTS: Of the 54 inguinal canal reoperations surgery was successful in all but 1 patient who sustained a transected vas deferens (1.8% complication rate). CONCLUSIONS: The surgical approach to the inguinal canal through the cremaster fascia is an effective technique for reoperation of the inguinal canal after orchiopexy or hernial repair. PMID- 11025755 TI - Paratesticular yolk sac tumor in a child. PMID- 11025756 TI - Contralateral testicular metastasis in paratesticular rhabdomyosarcoma. PMID- 11025757 TI - Monfort abdominoplasty with neoumbilical modification. AB - PURPOSE: In patients who undergo the Monfort abdominoplasty the umbilicus is placed high on the abdominal wall. We describe a technique for abdominal wall reconstruction in patients with the prune-belly syndrome with the modification of neoumbilicus creation at an anatomically normal position. MATERIALS AND METHODS: Five patients underwent modified Monfort abdominal wall reconstruction with the neoumbilicus created from an island flap of skin based on the fascial plate at the level of the iliac crest. RESULTS: All patients and parents were satisfied with the outcome. In addition to abdominoplasty, orchiopexy and ureteral reconstruction were also safely and easily performed. CONCLUSIONS: Neoumbilicus construction during Monfort abdominoplasty is an excellent approach to a more normal anatomical outcome. PMID- 11025758 TI - Fibronectin as a potent inhibitor of calcium oxalate urolithiasis. AB - PURPOSE: Fibronectin (230 kD.) is a multifunctional alpha2-glycoprotein distributed throughout the extracellular matrix and body fluids. Many investigators have demonstrated that fibronectin, because of its cell adhesive action, is related to biological processes such as morphogenesis, wound healing and metastasis. Recent studies have shown that a variety of molecules, including fibronectin, inhibit endocytosis of calcium oxalate crystals in vitro. We investigated other roles of fibronectin in calcium oxalate stone formation. MATERIALS AND METHODS: Immunoblotting of the crystal surface binding substance obtained from pooled healthy male urine samples was used to analyze whether fibronectin was adsorbed onto the surface of calcium oxalate crystals. To clarify the relationship between fibronectin and calcium oxalate crystals, we performed 6 experiments. Experiment 1 was immunohistochemical examination of fibronectin expression in stone forming rat model kidneys, and experiment 2 examined the fibronectin content of stone forming rat kidney models with the enzyme-linked immunosorbent assay. Experiment 3 was designed to determine fibronectin content of Madin-Darby canine kidney (MDCK) cells stimulated by addition of calcium oxalate crystals and experiment 4 identified the inhibitory effect of fibronectin on calcium oxalate crystal growth by the seed crystal method. For experiment 5 we used an aggregometer system to clarify the inhibitory effect of fibronectin on calcium oxalate crystal aggregation and experiment 6 examined the inhibitory effect of fibronectin on the adhesion of calcium oxalate crystals to MDCK cells. RESULTS: In the crystal surface binding substance immunoreactive bands at 230 kD., which correspond to the molecular weight of fibronectin, were detected by Western blot analysis. In stone forming rat kidneys strong expression of fibronectin was found on the renal tubules to which the crystals were attached. The fibronectin content of these kidneys was significantly greater than that of kidneys without calcium oxalate crystals. The fibronectin content of MDCK cells tended to increase in proportion to the concentration of calcium oxalate crystals added to the culture medium. The growth inhibition assay showed that the inhibitory effect of fibronectin on calcium oxalate crystal growth was small in relation to the quantity of fibronectin excreted. However, fibronectin had inhibitory effects on calcium oxalate crystal aggregation and adhesion of the crystals to MDCK cells. CONCLUSIONS: Fibronectin secretion can be stimulated by calcium oxalate crystals, and this protein, which is excreted from the tubular cells, may inhibit calcium oxalate crystal aggregation and attachment to cells. PMID- 11025759 TI - Transport of oxalate across the rabbit papillary surface epithelium. AB - PURPOSE: The renal papilla and pelvic recesses of the mammalian kidney are thought to be the anatomical sites of initial kidney stone formation. We studied oxalate transport across the renal papillary surface epithelium. MATERIALS AND METHODS: In vitro oxalate transport across the papillary surface epithelium was studied by dissecting it off of the papilla and mounting it in a modified Ussing chamber. Experiments were conducted using physiological Ringer's solution at 37C. Removal or appearance of radioactive oxalate from the solution bathing the apical papillary surface epithelium surface was used as a measure of transepithelial transport. RESULTS: When only the papillary surface epithelium apical surface was exposed to oxalate (17 micromol./l.), the apical to basolateral flux of oxalate was 7.1 +/- 1.5 nmol./cm.2 per hour. When only the basolateral side was perfused with oxalate, the basolateral to apical oxalate flux was 1.6 +/- 0.7 nmol./cm.2 per hour. When the apical and basolateral surfaces were exposed to oxalate, a net apical to basolateral flux of oxalate was 3.0 +/- 0.7 nmol./cm.2 per hour. There was almost complete inhibition of apical to basolateral oxalate transport when the experiments were conducted at 20C. Absorption of oxalate across the papillary surface epithelium was decreased by the addition of 1 mM. 4, 4' diisothiocyanatsostilbene-2,2'-disulphonic acid (DIDS), independent of sodium, but reduced by the presence of chloride in the apical solution. Increasing the pH of the apical solution from 7. 4 to 7.8 also resulted in a significant decrease in papillary surface epithelium oxalate uptake. CONCLUSIONS: Rabbit papillary surface epithelium demonstrates oxalate absorption by mechanisms that may be similar to those described previously for renal and colonic epithelia. Absorption of oxalate across the papillary surface epithelium may have a role in the prevention of initial calcium oxalate stone formation within the pelvic recesses. PMID- 11025760 TI - Interaction of nitric oxide and transforming growth factor-beta1 induced by angiotensin II and mechanical stretch in rat renal tubular epithelial cells. AB - PURPOSE: Changes in intrarenal pressure accompanying unilateral ureteral obstruction can result in tubular mechanical stretch and mediator release from renal tubules. Therefore, we examined the synthesis of nitric oxide and transforming growth factor-beta (TGF-beta), and their interaction in rat renal epithelial cells (NRK-52E) exposed to either angiotensin II or mechanical stretch. MATERIALS AND METHODS: NRK-52E were exposed to either angiotensin II or mechanical stretch. Nitrite and TGF-beta in the supernatant were assessed by the Greiss reaction and bioassay, respectively. The level of cell hypertrophy and intracellular TGF-beta protein was determined by flow cytometry. TGF-beta messenger RNA and inducible nitric oxide synthase protein were detected by reverse transcriptase polymerase chain reaction and Western blot, respectively. RESULTS: Angiotensin II stimulated TGF-beta1 and nitric oxide. The nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NAME) or angiotensin II type I receptor blocker, losartan, inhibited nitric oxide and TGF-beta1 induced by angiotensin II. Flow cytometry showed that either L-NAME or losartan inhibited angiotensin II induced cell hypertrophy. TGF-beta1 inhibited iNOS protein and nitric oxide, whereas an anti-TGF-beta antibody enhanced iNOS. Mechanical stretch induced TGF beta, inducible NOS protein and nitric oxide. However, TGF-beta expression was not affected by L-arginine or L-NAME when cells were exposed to mechanical stretch. CONCLUSIONS: These results demonstrate that nitric oxide is an intermediate in angiotensin II stimulated TGF-beta1 expression but not in stretch induced TGF-beta expression, and that TGF-beta1 is a negative regulator of nitric oxide in rat renal epithelial cells. The complex interaction of these cytokines may be a target for intervention in the fibrotic and apoptotic processes in the obstructed kidney. PMID- 11025761 TI - Quantification of the tip movement of lithotripsy flexible pneumatic probes. AB - PURPOSE: We developed an optical system to quantify in vitro tip movement of the Lithoclastdouble dagger flexible probe, and correlated various physical parameters of the vibrating probe tip with resultant stone fragmentation. MATERIALS AND METHODS: A noncontact optical measurement system was developed to quantify in vitro tip movement of the Lithoclast flexible probe. This system and an in vitro fragmentation model were used to determine the tip displacement, velocity, impact momentum, impact energy and stone fragmentation of the flexible probe at 5 deflection angles between 0 and 48 degrees, and the 2 pneumatic pressure levels of 2.0 and 2.5 bar. RESULTS: An increase in maximum probe tip displacement, velocity, impact momentum and energy, and stone fragmentation was seen as the pneumatic pressure was increased from 2.0 to 2.5 bar. A progressive decrease in these parameters was demonstrated as the probe tip was deflected, especially at deflection angles greater than 24 degrees. Impact momentum appears to be the physical parameter most closely correlated with stone fragmentation efficiency. CONCLUSIONS: The optical measurement system and in vitro fragmentation model developed allow one to quickly and reliably assess the performance of flexible pneumatic probes in vitro. This system can be used for general bench testing and basic research that can provide critical information for the design of more effective and efficient flexible pneumatic lithotripsy probes. PMID- 11025762 TI - State dependent response of the locus caeruleus neurons to bladder distention. AB - PURPOSE: The precise mechanism by which normal persons wake upon urinary sensation is unclear. The locus caeruleus in the pons is suggested to be involved in the activating systems for arousal. We evaluated the effects of bladder distention on the neural activity in the l. caeruleus under different states of anesthesia. MATERIALS AND METHODS: Experiments were performed on 20 male Sprague Dawley rats. The spontaneous discharge rate of single neurons in and around the l. caeruleus was recorded with the rat under urethane anesthesia administered intraperitoneally, while the electroencephalogram was monitored simultaneously. The changes of the discharge rate of the neurons were observed during saline infusion into the bladder or during tail pinch stimulation. After recording the location of the neurons was confirmed histologically. RESULTS: Of 42 l. coeruleus neurons whose response was examined during deep anesthesia 31 showed an excitatory response to bladder distention, followed by a change in electroencephalogram pattern to a faster and smaller rate, while the remaining 11 showed no response. Of 28 l. coeruleus neurons examined during light anesthesia only 1 neuron was excited and the remaining 27 showed no response. Virtually all of the l. coeruleus neurons unresponsive to bladder distention were responsive to tail pinch stimulation. Of the nonnoradrenergic neurons ventromedial to the l. coeruleus 4 of 7 during deep anesthesia and 5 of 8 during light anesthesia showed an excitatory response to distention. CONCLUSIONS: The excitatory response of the l. coeruleus neurons to bladder distention was strongly affected by the state of anesthesia. The response was observed only during deep anesthesia and was accompanied by lightening of the anesthesia. L. coeruleus may be involved in arousal which is mediated by bladder distention. PMID- 11025763 TI - Radiotelemetered cystometry in pigs: validation and comparison of natural filling versus diuresis cystometry. AB - PURPOSE: Cystometry has previously been performed in awake animals using vesical and abdominal catheters, and artificial bladder filling. Conventional urodynamic recordings may be obtained in this manner, albeit under nonphysiological and potentially stressful conditions. Therefore, we developed a technique to perform continuous, ambulatory cystometric monitoring in pigs. MATERIALS AND METHODS: A dual pressure radiotelemetry device was surgically implanted in 22 large white pigs. Vesical and abdominal pressures were recorded and validated, subtracted detrusor pressures were derived and natural fill and diuresis cystometry was compared. RESULTS: Continuous recordings were obtained for 1 to 24 hours, and the devices remained in the animals for up to 3 months. There were few complications and incrustation of the intravesical catheter tip occurred but it did not appear to affect recorded pressures. The pressure data were validated by comparison with filling pressures during bladder distention and simultaneous conventional cystometry at the end of the experimental period. Comparison of natural filling and diuresis cystometrograms showed that natural bladder filling results in higher maximum detrusor pressure during voiding (38.1 versus 33.9 cm. H2O, p <0.05), higher detrusor pressure after contractions (42.6 versus 32.2 cm. H2O, p <0.05) and more frequent detection of unstable contractions in pigs with detrusor instability secondary to experimental manipulation of the lower urinary tract (77.8% versus 45.0%, p <0.05). CONCLUSIONS: This technique allows continuous cystometric monitoring in less stressed animals under more physiological conditions for relatively long periods and, thus, allows prolonged assessment of bladder function in pigs in response to pathological and pharmacological manipulations. Nonphysiological rates of bladder filling have been shown to result in detrusor inhibition, which emphasizes the importance of ambulatory cystometry when describing bladder function. PMID- 11025764 TI - Similarity of distributions of spinal c-Fos and plasma extravasation after acute chemical irritation of the bladder and the prostate. AB - PURPOSE: Persistent pain in referred areas and voiding dysfunction are characteristic symptoms of chronic abacterial prostatitis. Since referred pain from visceral organs is considered a neurological event, it appeared reasonable to hypothesize that the persistent pain associated with prostatitis might also be explained by neural mechanisms. Neurogenic plasma extravasation and c-fos expression in the spinal cord, after chemical irritation of the rat prostate, was identified as a method to investigate the neurogenic aspect of prostatic inflammation. MATERIALS AND METHODS: The distribution of plasma extravasation using Evans blue dye was determined after chemical irritation of the prostate and bladder of the rat, and the distribution of dye extravasation was analyzed. c-fos expression within the spinal cord was determined immunocytochemically after chemical irritation of the prostate, bladder and superficial somatic region determined by the dye extravasation as a referred pain area (tail root). RESULTS: Chemical irritation of the prostate resulted in plasma extravasation in L5 to S2 dermatomes (primarily in L6 and S1). In rats receiving bladder irritation, the distribution of plasma extravasation showed a similar pattern to that observed in animals receiving prostatic irritation. Chemical irritation of the 3 structures resulted in expression of c-fos positive cells within the lumbosacral spinal cord. With each treatment the majority of c-fos positive cells were in the L6 and S1 segments. In all 3 groups the highest percentages of c-fos positive cells were observed in deeper laminae, including the dorsal commissure and sacral parasympathetic nucleus. CONCLUSIONS: Our results strongly suggest that referred pain status in inflammation of the bladder and prostate is neurogenically mediated. Based on these studies, there should be significant overlaps of nociceptive neurons within the spinal cord, which receive nociceptive inputs from pelvic soma and viscera. PMID- 11025765 TI - Up-regulation of bradykinin response in rat and human bladder smooth muscle. AB - PURPOSE: Responses to bradykinin were investigated in vitro in isolated control and hypertrophic smooth muscle strips from rat bladder. MATERIALS AND METHODS: Bladder hypertrophy was induced by a 10-day period of partial urinary outflow obstruction. In addition, human bladder strips were also investigated. RESULTS: Bradykinin (1 nM. to 1 microM.) caused contractions in all tissues studied. In the freshly isolated rat bladder preparations bradykinin induced contractions were similar and of small amplitude in control and hypertrophic tissues. After a 4-hour equilibratory period contractile responses to bradykinin and the B1 specific bradykinin receptor agonist desArg9 bradykinin were slightly increased in the controls but there was approximately a 6-fold increase in the hypertrophic muscle strips. After 4 hours of equilibration the human bladder strips showed a smaller but still significant increase in contractile response to bradykinin. Indomethacin, a cyclooxygenase inhibitor, almost abolished the increased response, which suggests that prostanoids are involved in the up-regulated response. The protein synthesis inhibitor cycloheximide inhibited up-regulation by approximately 50% in hypertrophic and control muscle strips from rat bladder and normal muscle from human bladder. CONCLUSIONS: These results demonstrate that bradykinin receptor responses are present in rat and human detrusor muscle and they can be up-regulated in vitro. Experiments on hypertrophic rat bladder revealed that this process is enhanced in hypertrophy. PMID- 11025766 TI - Protein kinase c regulates purinergic component of neurogenic contractions in mouse bladder. AB - PURPOSE: We evaluate the role of protein kinase C in excitatory purinergic neurotransmission in the mouse bladder. MATERIALS AND METHODS: In isolated mouse detrusor strips contractile responses to electrical field stimulation were mostly mediated by neural released acetylcholine and adenosine triphosphate (ATP). The changes in neurotransmission were measured indirectly by recording the contraction of detrusor strips in response to repetitive electrical field stimulation by trains of electrical pulses at 8 Hz. 1 second in duration. RESULTS: A protein kinase C activator, 1 to 2.5 nM. (beta-phorbol-12,13 dibutyrate (beta-PDBu), but not the inactive form alpha-phorbol-12,13-dibutyrate, significantly enhanced neurogenic detrusor contractions. The purinergic component of the evoked detrusor contractions in the presence of atropine was specifically sensitive to this enhancing effect by beta-PDBu but the cholinergic component in the alpha,beta-methylene ATP treated detrusors remained unaffected. This enhancing effect of beta-PDBu was dependent on the extracellular calcium (Ca2+) concentration. A P and/or Q type Ca2+ channel blocker, 0.1 and 0.3 microM. omega conotoxin-MVIIC, and protein kinase C inhibitors, 0.3 and 1 microM. staurosporine and 0.3 and 1 microM. bisindolylmaleimide I but not 0.1 and 0.3 microM. omega conotoxin-GVIA, an N type Ca2+ channel blocker, abolished the effect of beta PDBu. Moreover, beta-PDBu did not affect the muscle responses induced by the exogenous agonists carbachol or alpha,beta-methylene ATP and potassium chloride. CONCLUSIONS: These results suggest that the activation of Ca2+ channel, especially the P and/or Q type, may be involved in the enhancing effect of protein kinase C activator beta-PDBu on muscle contractions elicited by excitatory purinergic neurotransmission in the mouse detrusor strips. PMID- 11025767 TI - A comparison of cytology and fluorescence in situ hybridization for the detection of urothelial carcinoma. AB - PURPOSE: We determine the relative sensitivities of cytology and fluorescence in situ hybridization (FISH) for the detection of urothelial carcinoma. MATERIALS AND METHODS: A mixture of fluorescent labeled probes to the centromeres of chromosomes 3, 7 and 17, and band 9p21 (P16/CDKN2A gene) was used to assess urinary cells for chromosomal abnormalities indicative of malignancy. A total of 280 urine specimens from 265 patients, including 150 with a history of urothelial carcinoma and 115 without a history of urothelial carcinoma, were analyzed. FISH analysis was performed without prior knowledge of clinical findings, that is biopsy, cystoscopy and cytology results. A positive result was defined as 5 or more urinary cells with gains of 2 or more chromosomes. RESULTS: A total of 75 biopsies showed urothelial carcinoma at FISH analysis among the 265 patients. The sensitivity of urine cytology for pTa (36 cases), pTis (18) and pT1-pT4 (15) tumors was 47%, 78% and 60%, respectively, for an overall sensitivity of 58%. The sensitivity of FISH for pTa (37 cases), pTis (17) and pT1-pT4 (19) tumors was 65%, 100% and 95%, respectively, for an overall sensitivity of 81%. FISH was significantly more sensitive than cytology for pTis (p = 0.046), pT1-pT4 (p = 0.025), grade 3 (p = 0.003) and all tumors (p = 0.001). The specificity of cytology and FISH among patients without cystoscopic evidence of urothelial carcinoma and no history of urothelial carcinoma was 98% and 96%, respectively (p = 0.564). CONCLUSIONS: The sensitivity of FISH for the detection of urothelial carcinoma is superior to that of cytology, and the specificity of FISH and cytology for urothelial carcinoma are not significantly different. Further prospective studies are required but FISH has the potential to improve significantly the management of urothelial carcinoma. PMID- 11025768 TI - The effect of halofuginone, an inhibitor of collagen type i synthesis, on urethral stricture formation: in vivo and in vitro study in a rat model. AB - PURPOSE: Urethral strictures are narrowing of the urethra caused by fibrosis due to excessive collagen production in response to an insult. We evaluated the effects of halofuginone, a potent inhibitor of collagen alpha1(I) gene expression, on experimentally induced urethral strictures in vivo and on rat urethral fibroblasts in vitro. MATERIALS AND METHODS: Applying coagulation current to the male rat urethra produced urethral strictures. Halofuginone was given to the animals for 7 days, starting on the day of stricture formation, either orally at 1 and 5 ppm in the diet or by injection of 0.03% halofuginone solution into the urethra. All rats were sacrificed on day 21. Collagen alpha1(I) gene expression was evaluated by in situ hybridization, collagen content by sirius red staining and urethral morphology by urethrogram. RESULTS: Coagulation current produced reproducible strictures with a typical urethrogram appearance, which were associated with increases in collagen alpha1(I) gene expression and collagen content at the stricture site. Halofuginone injected into the urethra or orally at 5 ppm normalized the urethrogram and prevented increases in collagen alpha1(I) gene expression and collagen content. Halofuginone at a concentration of 10-8 M. inhibited the collagen secreted by fibroblasts derived from the rat male urethra, which was due to inhibition of the collagen alpha1(I) gene expression. CONCLUSIONS: Halofuginone prevented stricture formation and may become an important mode of therapy in the prevention of restenosis during urethral stricture formation. PMID- 11025769 TI - Age dependent apoptosis and loss of rhabdosphincter cells. AB - PURPOSE: To our knowledge the exact age dependent morphological and functional changes of the sphincter mechanism have not been investigated. Therefore, cell densities of the urethra and the urethral rhabdosphincter across various age groups, and the appearance of apoptosis were examined to explore the changes in these structures during the aging process. MATERIALS AND METHODS: Specimens were obtained from 16 male and 7 female cadavers 5 weeks to 92 years old. Histological sections were taken from 3 different levels of the rhabdosphincter and urethra. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method was used to detect apoptosis in the urethra and rhabdosphincter. In all specimens relative volume densities of the striated muscle fibers, apoptotic indexes and diameters of the rhabdosphincter and urethra were determined. RESULTS: An age dependent increase of apoptosis of the striated muscle fibers of the rhabdosphincter led to a dramatic decrease in the number of striated muscle cells. In the 5-week-old neonate 87.6% and in the 91-year-old woman 34.2% of the rhabdosphincter consisted of striated muscle cells. Overall, a direct linear correlation between the age of the specimens and decrease in volume densities of the striated muscle cells was evident. CONCLUSIONS: The dramatic decrease in the number of striated muscle cells in the rhabdosphincter of the elderly due to apoptosis represents the morphological basis for the high incidence of stress incontinence in this population. PMID- 11025770 TI - Urethral development in the fetal rabbit and induction of hypospadias: a model for human development. AB - PURPOSE: To determine whether the development of the rabbit phallus would be an appropriate model of human phallic development, we evaluated the formation of the fetal rabbit phallus and attempted to induce hypospadias pharmacologically. MATERIALS AND METHODS: New Zealand rabbit fetuses were obtained on gestational days 20 to 24, 26, 28 and 31. Sex was determined by gonadal morphology, and 6 fetuses were obtained at each age. The perineum was dissected, fixed, sectioned and stained with hematoxylin and eosin, and monoclonal antibodies against neuronal specific enolase. Two pregnant rabbits were treated with 10 mg./kg. finasteride orally daily between gestational days 19 and 28. The development of the external genitalia was compared in treated and untreated control rabbits. RESULTS: The rabbit phallus contains 2 corpora cavernosa and dorsolateral nerves similar to the human. In male and female fetuses fusion of the urethral folds progressed in a proximal to distal sequence forming a seam at the point of ventromedial fusion. In male fetuses urethral fold and ventral preputial fusion continued more distally toward the glans compared to females. Thus, in mature males the urethral meatus and ventral prepuce extended to the tip of the phallus, whereas in females the urethral meatus opened on the proximal phallus and the prepuce was deficient ventrally forming a dorsal hood. Male offspring had a significantly larger anogenital distance postnatally than female offspring. In male fetuses exposed to finasteride urethral fusion did not extend distally and the prepuce was deficient ventrally. Also, male offspring exposed to finasteride in utero had a significantly shorter anogenital distance than females and untreated control males at all ages (p <0.05). CONCLUSIONS: Fetal development of the rabbit phallus and urethra is homologous to the human. Although the gestational period is significantly shorter, the temporospatial pattern of external genitalia development is analogous in these species. Feminization of the rabbit urethra, hypospadias, can be induced by inhibiting 5alpha-reductase. Use of this animal model will allow further study of molecular mechanisms involved in urethral fusion and the evaluation of the pathophysiological processes of hypospadias. PMID- 11025771 TI - Increases in intracavernous penile pressure following injections of excitatory amino acid receptor agonists in the hypothalamic paraventricular nucleus of anesthetized rats. AB - PURPOSE: We examined whether N-methyl-D-aspartate (NMDA), amino-3-hydroxy-5 methyl-isoxazole-4-propionic acid (AMPA) or trans-1-amino-1,3-cyclo pentadicarboxylic acid (ACPD) increase intracavernous pressures when injected in the paraventricular nucleus of the hypothalamus. MATERIALS AND METHODS: Sprague Dawley rats weighing 250 to 300 gm. were anesthetized with pentobarbital and placed in a stereotaxic apparatus after catheterization of the carotid artery and insertion of a 25 gauge needle in the corpus cavernosum. Both catheters were coupled to pressure transducer systems. Electrical stimulations with glass tungsten microelectrodes and chemical injections with a 0.5 microl. syringe were done in the paraventricular nucleus. At the end of each experiment rats were perfused with saline and 10% formalin solutions. Brains slices 16 microm. thick were mounted and stained with cresyl violet to identify chemical and electrical stimulation sites. RESULTS: Electrical stimulations of the paraventricular nucleus increased intracavernous pressure from 15.5 +/- 5.7 to 50.02 +/- 20.4 cm. H2O in 10 rats (p <0.001) and decreased central pressure in 10 (p <0. 001). Intracavernous pressure increased from 17.2 +/- 5.26 to 60.6 +/- 14.36 cm. H2O with NMDA, 26.0 +/- 8.9 to 49.6 +/- 12.2 cm. H2O with AMPA and 18.0 +/- 2.3 to 41.0 +/- 22.68 cm. H2O with ACPD when injected in the paraventricular nucleus of 5 rats (analysis of variance 12.18, df 3, p <0.007). Mean delays and durations varied between 3.6 to 4.0 and 5.6 to 7.0, respectively, and were similar for all agonists (analysis of variance not significant). CONCLUSIONS: Glutamate receptor subtypes NMDA, ACPD and AMPA cause an increase in intracavernous pressure when injected in the paraventricular nucleus. Therefore, increases in intracavernous pressure following injection of glutamate must be initiated by the synergistic or additive effects of all of these receptor subtypes. PMID- 11025772 TI - Effect of a Chinese herbal medicine mixture on a rat model of hypercholesterolemic erectile dysfunction. AB - PURPOSE: We examine the effect of a Chinese herbal medicine mixture on erectile function in a rat model of hypercholesterolemic erectile dysfunction. MATERIALS AND METHODS: In this study 32, 3-month-old Sprague-Dawley rats were used. The 8 control animals were fed a normal diet and the remaining 24 were fed 1% cholesterol diet for 4 months. After 2 months herbal medicine was added to the drinking water of the treatment group of 16 rats but not the cholesterol only group of 8. Of the 16 rats 8 received 25 mg./kg. per day (group 1) and 8 received 50 mg./kg. per day (group 2) of Chinese herbal medicine mixture. Serum cholesterol levels were measured at 2 and 4 months. At 4 months erectile function was evaluated with cavernous nerve electrostimulation in all animals. Penile tissues were collected for electron microscopy, and to perform Western blot for endothelial nitric oxide synthase, neuronal nitric oxide synthase, basic fibroblast growth factor (bFGF) and caveolin-1. RESULTS: Serum cholesterol levels were significantly higher in animals fed the 1% cholesterol diet compared to controls at 2 and 4 months. Nevertheless, there was no significant difference among group 1 (145 +/- 30 mg./dl.), group 2 (157 +/- 20) and the cholesterol only group (143 +/- 15). Systemic arterial pressure was not significantly different between the animals that were fed the 1% cholesterol diet and the controls. During electrostimulation of the cavernous nerve peak sustained intracavernous pressure was significantly lower in the cholesterol only group (50 +/- 23 cm. H2O) compared to the control group. Conversely erectile function was not impaired in the herbal medicine treated rats. Electron microscopy showed many caveolae with fingerlike processes in the cavernous smooth muscle and endothelial cell membranes in control and treated rats but not in the cholesterol only group of rats. Western blot did not show a difference among groups in protein expression for endothelial nitric oxide synthase and neuronal nitric oxide synthase in penile tissue but caveolin-1 and bFGF protein expression was significantly higher in groups 1 and 2 than in the cholesterol only and control groups. CONCLUSIONS: Rats developed erectile dysfunction after being fed a 1% cholesterol diet for 4 months. Although serum cholesterol levels were similar in the cholesterol only rats and those treated with Chinese herbal medicine mixture, erectile response was significantly better in the treated group. The mechanism of the herbal medicine is unknown. High levels of bFGF and caveolin-1 expression in the treated group may protect the cavernous smooth muscle and endothelial cells from the harmful effect of high serum cholesterol. PMID- 11025773 TI - Organization and relative content of smooth muscle cells, collagen and elastic fibers in the corpus cavernosum of rat penis. AB - PURPOSE: The corpus cavernosum smooth muscle and extracellular matrix are essential for normal penile erection and are implicated in erectile dysfunction. Although investigations of these issues have used the rat corpus cavernosum, organization of its components is to date not well known. We characterized and quantified the smooth muscle cells and the main extracellular matrix components of the rat corpus cavernosum. MATERIALS AND METHODS: Collagen, elastic fibers and smooth muscle cells were stained on paraffin sections of rat penises using sirius red and Gomori's reticulin, Weigert's resorcin-fuchsin and an anti-smooth muscle cells alpha-actin antibody, respectively. Stained components were then quantified by computer aided morphometry. RESULTS: Smooth muscle cells were restricted to the subendothelial space of corpus cavernosum and had a volumetric density of 9.1%. Collagen was thick, usually in transversely oriented bundles and was the most abundant component of the trabeculae with a volumetric density of 62.7%. Gomori's reticulin disclosed a meshwork of fibrils also in the subendothelial space but did not stain the thicker bundles. Volumetric density of elastic fibers was 4.9%, and at the periphery of the corpus cavernosum the fibers were parallel to the long axis of the penis, while in deeper regions most of them were transversely oriented and at different directions from those of collagen. CONCLUSIONS: Rat corpus cavernosum differs from that of humans by lesser amounts of smooth muscle cells, greater amounts of collagen and the presence of fibrillar collagen and smooth muscle cell subendothelial layers. Therefore, these differences should be considered when using the rat penis for studies on erection. PMID- 11025774 TI - Reduction of endothelial and smooth muscle density in the corpora cavernosa of the streptozotocin induced diabetic rat. AB - PURPOSE: Erectile dysfunction is one of the most prevalent complications of diabetes in males. Because adequate vascular perfusion is needed for appropriate erectile tissue function a likely reason for the high incidence of this complication in diabetics is a pathological change associated with the disease in vascularization of erectile tissues. We investigate whether chronic diabetes may induce changes in vascularization of the corpora cavernosa using a computerized image analysis system to quantify changes in the smooth muscle and endothelial cell content of the corpora cavernosa of diabetic rats induced by streptozotocin 6 months previously, and compare these changes to those associated with aging. MATERIALS AND METHODS: We studied 3 groups of rats, including 10-week-old untreated controls, diabetic rats treated with streptozotocin for 6 months starting at age 10 weeks and 18-month-old rats (aged). Penile shafts from these groups were excised, fixed, sectioned and immunostained with anti-smooth muscle actin to identify smooth muscle cells and anti-CD31 to identify endothelial cells. Computerized image analysis was used to quantify the percent area within the corpora cavernosa occupied by smooth muscle cells or endothelial cells, and the data were compared among the groups. RESULTS: We identified a highly significant decrease in the percentage of smooth muscle and endothelial cells within the cavernosa areas of diabetic rats compared to control or aged rats. Mean cavernous smooth muscle cell content was 15.28 +/- 2.54% in control rats and 9.83 +/- 1.21% in diabetic rats (p = 0.0001). Likewise, cavernous endothelial cell content was 6.93 +/- 0.86% in the control group and 4.01 +/- 1.08% in the diabetic group (p = 0. 0001). However, no statistical difference of smooth muscle or endothelial cell content was found between control and aged rats. CONCLUSIONS: Using the streptozotocin treated rat as a model for diabetes, we showed that smooth muscle and endothelial cell density is significantly decreased in diabetic corpora cavernosa but not in normal aged rats. This observation is a further step toward the understanding of the pathomechanisms for diabetic related erectile dysfunction. PMID- 11025775 TI - In vitro and in vivo effects of vitamin D (calcitriol) administration on the normal neonatal and prepubertal prostate. AB - PURPOSE: Although many studies have investigated the role of calcitriol in the growth regulation of normal and cancerous prostates, little is known about its role in early prostatic development. The interactions between calcitriol and androgens, and their actions on the normal prostate have similarly been proposed but not evaluated. Previous studies in our laboratory have revealed that in utero administration of 1,25-dihydroxycholecalciferol or calcitriol can influence prostate growth and differentiation throughout the life of the animal. We further examined the influence of calcitriol on the normal prostate in vitro and in vivo by focusing on early stages of prostatic development. MATERIALS AND METHODS: The effects of calcitriol on the growth of the normal human neonatal prostatic epithelial cell line 267B-1 was determined in the presence and absence of dihydrotestosterone (DHT). We also examined the effect of calcitriol on the growth of maturing rat prostates in vivo. Before puberty 4 groups of rats 27 to 38 days old were treated with vehicle (controls) or calcitriol. When the rats reached adulthood at age 100 to 110 days a control group and a calcitriol group were sacrificed. The other 2 groups were given exogenous DHT for 5 days. For the animals to become adapted to DHT they were kept alive for 1 additional week and sacrificed at about age 120 days. RESULTS: In vitro studies demonstrated that 267B-1 cells possess vitamin D receptors and their growth was inhibited by calcitriol with an IC50 (concentration resulting in 50% cytotoxicity) of 30 microM. Proliferation of these neonatal prostate cells was also inhibited by calcitriol in the presence of DHT in vitro. Our studies indicate that, although calcitriol was administered at the apparently important prepubertal period, there was no difference in prostatic weights between the control and calcitriol treated rats. Exogenous administration of DHT decreased prostatic weight of control rats but in rats treated with 1,25-dihydroxycholecalciferol DHT did not have any significant effect on prostatic weight. No statistically significant differences were observed in seminal vesicle weights among the different groups of animals. Analysis of the nuclear matrix protein composition of the prostatic tissue showed differences in composition between the DHT, and calcitriol and DHT treated rat prostates. CONCLUSIONS: These studies indicate that calcitriol administered just before puberty does not significantly influence prostatic growth in the presence of endogenous or exogenous administered DHT, and has an inhibitory effect on neonatal prostate epithelial cell growth in vitro in the presence and absence of DHT. Treatment with calcitriol and DHT also results in differences in nuclear matrix protein composition. Prepubertal administration of calcitriol may inhibit the exogenous DHT action in decreasing epithelial growth and stimulating stromal proliferation in the rat prostate. PMID- 11025776 TI - PSP94 expression after androgen deprivation therapy: a comparative study with prostate specific antigen in benign prostate and prostate cancer. AB - PURPOSE: To examine the clinical use of PSP94 (prostate secretory protein of 94 amino acids) as an androgen independent marker, we conducted a comparative study of prostate samples including benign tissue and cancers which did and did not have androgen deprivation. MATERIALS AND METHODS: Among 163 radical prostatectomy cases 75 had androgen deprivation before operation, while surgery was performed in the remainder without prior hormone treatment. Considering the pathological up grading following hormone therapy, contiguous sections from radical prostatectomy samples were stained for PSP94 and prostate specific antigen (PSA) by immunohistochemistry, and equivalent tumor foci were evaluated by assessing the intensity and extent of the staining. RESULTS: In untreated benign prostate tissue PSP94 and PSA staining was positive and identical in all sections in the no pretreatment group. However, PSP94 expression in the androgen deprivation group was significantly higher than PSA in intensity (p = 0.0005) and extent (p = 0.034). In untreated cancer cases PSP94 intensity and extent demonstrated strong inverse association with Gleason grade (p <0.0001). In contrast, PSA expression was high in every grade, resulting in no statistical association with tumor grade. In the androgen deprivation group PSA staining was decreased in every grade compared to the no pretreatment group. On the other hand, PSP94 expression was decreased in grade 3 tumor foci but increased in grades 4 and 5 tumor foci compared with samples of the corresponding grade in the no pretreatment group (p = 0.0034). CONCLUSIONS: PSP94 expression in benign prostate persists under androgen deprivation compared to PSA. PSP94 synthesis in high grade tumor appears to be activated in the absence of androgen stimulation, indicating the possible alternative pathways in the regulation of PSP94. PMID- 11025777 TI - Antibody immunity to prostate cancer associated antigens can be detected in the serum of patients with prostate cancer. AB - PURPOSE: Several immune based therapies targeting prostate cancer associated proteins are currently undergoing clinical investigation. In general, however, little is known about the immunogenicity of prostate cancer or which prostate cancer associated proteins elicit immune responses. We determine whether patients with prostate cancer have antibody immunity to known prostate cancer associated proteins, what the prevalence of this immunity is and whether immunity to individual proteins is associated with the stage of disease. MATERIALS AND METHODS: We evaluated the inherent humoral immune response against prostate specific antigen (PSA), prostatic acid phosphatase, p53 and HER-2/neu, all known prostate cancer associated proteins, in 200 patients with various stages of disease and male controls. RESULTS: Antibody immunity to PSA was significantly different between the patient (11%, 22 of 200) and control populations (1.5%, 3 of 100, p = 0.02), and titers 1:100 or greater were particularly prevalent in the subgroup of patients with androgen independent disease (11%, 6 of 56). Antibody immunity to prostatic acid phosphatase and p53 was detected (5.5%, 11 of 200 and 6%, 12 of 200), and was not different from the control population (4%, 4 of 100, p = 0.57 and 7%, 7 of 100, p = 0.74). Antibody immunity to HER-2/neu was significantly higher in patients with prostate cancer (15.5%, 31 of 200) compared to controls (2%, 2 of 100, p = 0.0004), and titers 1:100 or greater were most prevalent in the subgroup of patients with androgen independent disease (16%, 9 of 56). CONCLUSIONS: These findings suggest that prostate cancer is an immunogenic tumor. Moreover, for PSA and HER-2/neu the prevalence of antibody immunity was higher in patients with androgen independent disease, indicating that even patients with advanced stage prostate cancer can have an immune response to their tumor. PMID- 11025778 TI - The mismatch repair gene hMSH2 is mutated in the prostate cancer cell line LNCaP. AB - PURPOSE: Mismatch repair genes are responsible for the coordinated correction of misincorporated nucleotides formed during DNA replication. Inactivating and inherited mutations in the prototypic mismatch repair gene hMSH2 have been described in a cancer predisposition syndrome known as hereditary nonpolyposis colon cancer. Patients with hereditary nonpolyposis colon cancer are at increased risk for colon cancer and extracolonic cancers such as upper tract transitional cell carcinoma but not prostate cancer. We investigated expression of hMSH2 in prostate cancer cell lines using genetic and molecular analysis. MATERIALS AND METHODS: We used the 3 well described prostate cancer cell lines, DU145, LNCaP and PC3. Western blot analysis with monoclonal antibody to hMSH2 was used to assess expression. Southern blot and polymerase chain reaction of genomic DNA were used to identify genetic alterations in the hMSH2 gene. Single cell cloning, dinucleotide repeats and BAT-26 were used to assess the cell lines for microsatellite instability. RESULTS: The prostate cancer cell line LNCaP did not express hMSH2 and was found to have a homozygous deletion of hMSH2 exons 9 to 16, resulting in truncation of the protein. While microsatellite analysis did not reveal alterations at the BAT-26 locus, single cell cloning produced several LNCaP subclones with alteration at 1 dinucleotide repeat. CONCLUSIONS: The well described prostate cancer cell line LNCaP has a mutation in the hMSH2 gene, resulting in loss of expression and possible evidence of microsatellite instability. To our knowledge our finding is the first demonstration of a genetic alteration in hMSH2 in a prostate cancer cell line. PMID- 11025779 TI - Immunomagnetic cell enrichment detects more disseminated cancer cells than immunocytochemistry in vitro. AB - PURPOSE: We describe a method to improve tumor cell detection compared to currently available immunocytochemical methods by using immunomagnetic cell enrichment. MATERIALS AND METHODS: Two different methods of immunomagnetic cell enrichment using antibody coated magnetic beads were tested and compared with unenriched immunocytochemistry. One method was positive selection of epithelial cells from mononuclear cells with the antiepithelial antibody BER-EP4 and the other was depletion of mononuclear cells with the antileukocyte antibody CD45. Mononuclear cells were isolated from peripheral blood by density centrifugation and various numbers of tumor cells were added. The 5 different cell lines from urological malignancies used in the study were DU-145, RT-4, CAKI-2, KTCTL-2 and KTCTL-30. Following incubation of cell suspensions with the beads, cell separation was performed in a magnetic field. After centrifugation on glass slides immunocytochemical staining for cytokeratin was performed. A total of 112 experiments were completed and negative controls were obtained. RESULTS: The number of tumor cells detected by positive selection and depletion was significantly higher than by immunocytochemistry (p <0.001). The median enrichment factor and tumor cell recovery rate for positive selection and depletion were 15.3 and 61.2%, and 13.0 and 57.3%, respectively (not significant). With less than 1 tumor cell suspended in 106 mononuclear cells, the probability of tumor cell detection was 23% for immunocytochemistry alone and 93.3% for both enrichment methods (p <0.01). No false-positive results were observed. CONCLUSIONS: Compared to immunocytochemistry, immunomagnetic cell enrichment significantly improves the sensitivity of detection of epithelial cells added to mononuclear cells. Both methods of enrichment were equally effective and may be important for clinical practice in the future. PMID- 11025780 TI - Electronic prescribing and monitoring are needed to improve drug use. PMID- 11025781 TI - Preventable medical injuries in older patients. AB - Injuries associated with hospitalization are more common in older (>/=65 years) than in younger patients (<65 years), and they may be more severe and more often preventable. The increasing age of the population magnifies the importance of this problem. In this review, we first consider medical injuries in general and then review the literature for 6 categories: adverse drug events, falls, nosocomial infections, pressure sores, delirium, and surgical and perioperative complications. For each of these categories, older patients appear to be at higher risk, ranging from a 2.2-fold increase for perioperative complications to a 10-fold increase for falling, based on Harvard Medical Practice Study rates. The main cause of these increased risks appears to be the diminished physiological reserve of elderly patients; however, age alone is a less important predictor of adverse events than comorbidities and functional status. Furthermore, many of these complications appear to be preventable, although the proportion preventable varies by type of complication. While some prevention strategies are specifically beneficial in older patients, many apply to all age groups. Geriatric care units and consultation systems have improved outcomes in some instances, although the data are mixed. The success of intervention varies by type of complications. For medications, various interventions have been successful, and fall prevention programs have been demonstrated to be effective in the nursing home and home. PMID- 11025782 TI - The diagnostic value of historical features in primary headache syndromes: a comprehensive review. AB - BACKGROUND: Existing criteria for the diagnosis of headache do not indicate which clinical features are most important to establish or exclude primary headache diagnoses. OBJECTIVE: To determine the value of history taking in the evaluation of patients with primary headache syndromes. METHODS: A MEDLINE search of English language studies published since 1966 and a hand search of bibliographies from retrieved articles. Included studies cited the sensitivity of clinical and historical features for the diagnosis of migraine, tension-type, and cluster headaches. Studies were classified according to their use of the International Headache Society criteria or other criteria as the reference standard for diagnosis. RESULTS: The features most predictive of migraine, when compared with tension-type headache, are nausea, photophobia, phonophobia, and exacerbation by physical activity. Positive likelihood ratios (95% confidence intervals) are 19.2 (15.0-24.5), 5.8 (5.1-6.6), 5.2 (4.5-5.9), and 3.7 (3.4-4.0), respectively. Headache precipitants are present equally in patients with migraine and tension type headache, with the exception of food triggers, which confer a positive likelihood ratio of 3.6 (2.8-4.6) for the diagnosis of migraine. Visual aurae are present in 84% of patients with migraine with aura. Cluster headache is strictly unilateral, is periorbital in 80% of patients, and lasts less than 1 hour in 54% of patients. CONCLUSIONS: The features most predictive of the diagnosis of migraine, when compared with tension-type headache, are nausea, photophobia, phonophobia, exacerbation by physical activity, and an aura. Cluster headache is a distinct clinical syndrome. PMID- 11025783 TI - Effects of computerized physician order entry on prescribing practices. AB - BACKGROUND: Computerized order entry systems have the potential to prevent errors, to improve quality of care, and to reduce costs by providing feedback and suggestions to the physician as each order is entered. This study assesses the impact of an inpatient computerized physician order entry system on prescribing practices. METHODS: A time series analysis was performed at an urban academic medical center at which all adult inpatient orders are entered through a computerized system. When physicians enter drug orders, the computer displays drug use guidelines, offers relevant alternatives, and suggests appropriate doses and frequencies. RESULT: For medication selection, use of a computerized guideline resulted in a change in use of the recommended drug (nizatidine) from 15.6% of all histamine(2)-blocker orders to 81.3% (P<.001). Implementation of dose selection menus resulted in a decrease in the SD of drug doses by 11% (P<.001). The proportion of doses that exceeded the recommended maximum decreased from 2.1% before order entry to 0.6% afterward (P<.001). Display of a recommended frequency for ondansetron hydrochloride administration resulted in an increase in the use of the approved frequency from 6% of all ondansetron orders to 75% (P<.001). The use of subcutaneous heparin sodium to prevent thrombosis in patients at bed rest increased from 24% to 47% when the computer suggested this option (P<.001). All these changes persisted at 1- and 2-year follow-up analyses. CONCLUSION: Computerized physician order entry is a powerful and effective tool for improving physician prescribing practices. PMID- 11025784 TI - Periodontal disease and risk of cerebrovascular disease: the first national health and nutrition examination survey and its follow-up study. AB - BACKGROUND: Periodontal disease has been found to be a potential risk factor for coronary heart disease. However, its association with cerebrovascular accidents (CVAs) is much less studied. METHODS: This study examines the association between periodontal disease and CVA. The study cohort comprises 9962 adults aged 25 to 74 years who participated in the First National Health and Nutrition Examination Survey and its follow-up study. Baseline periodontal status was categorized into (1) no periodontal disease, (2) gingivitis, (3) periodontitis, and (4) edentulousness. All CVAs (International Classification of Diseases, Ninth Revision [ICD-9], codes 430-438) were ascertained by hospital records for nonfatal events and death certificates for fatal events. The first CVA, nonfatal or fatal, was used to define incidence. Relative risks were estimated by hazard ratios from the Cox proportional hazard model with adjustment for several demographic variables and well-established cardiovascular risk factors. Weights were used to generate risk estimates. RESULTS: Periodontitis is a significant risk factor for total CVA and, in particular, nonhemorrhagic stroke (ICD-9, 433 434 and 436-438). Compared with no periodontal disease, the relative risks (95% confidence intervals) for incident nonhemorrhagic stroke were 1.24 (0.74-2.08) for gingivitis, 2.11 (1.30-3.42) for periodontitis, and 1.41 (0.96-2.06) for edentulousness. For total CVA, the results were 1.02 (0.70-1.48) for gingivitis, 1.66 (1.15-2.39) for periodontitis, and 1.23 (0.91-1.66) for edentulousness. Increased relative risks for total CVA and nonhemorrhagic stroke associated with periodontitis were also seen in white men, white women, and African Americans. Similar results were found for fatal CVA. CONCLUSION: Periodontal disease is an important risk factor for total CVA and, in particular, nonhemorrhagic stroke. PMID- 11025785 TI - Prevalence and phenotypic distribution of dyslipidemia in type 1 diabetes mellitus: effect of glycemic control. AB - BACKGROUND: Data on the prevalence of dyslipidemia in type 1 diabetes mellitus are scarce and are based on total triglyceride and total cholesterol concentrations alone. OBJECTIVE: To assess the effect of glycemic optimization on the prevalence of dyslipidemia and low-density lipoprotein cholesterol (LDL-C) concentrations requiring intervention in patients with type 1 diabetes. PATIENTS: A total of 334 adults with type 1 diabetes and 803 nondiabetic control subjects. METHODS: Levels of glycosylated hemoglobin, total cholesterol, total triglyceride, high-density lipoprotein cholesterol (HDL-C), and LDL-C were assessed at baseline and after 3 to 6 months of intensive therapy with multiple insulin doses. RESULTS: Levels of LDL-C greater than 4.13 mmol/L (>160 mg/dL) and total triglyceride greater than 2.25 mmol/L (>200 mg/dL) and low HDL-C levels (<0.9 mmol/L [<35 mg/dL] in men or <1.1 mmol/L [<45 mg/dL] in women) were found in 16%, 5%, and 20% of patients and 13%, 6%, and 9% of controls, respectively (P<.001 for HDL-C). Diabetic women showed more hypercholesterolemia than nondiabetic women (15.6% vs 8.5%; P =.04). After glycemic optimization (mean +/- SD glycosylated hemoglobin decrease, 2.2 +/- 1.96 percentage points), the prevalence of LDL-C levels greater than 4.13 mmol/L (>160 mg/dL) became lower in diabetic men than in nondiabetic men (9.7% vs 17.5%; P =.04), but women showed frequencies of dyslipidemia similar to their nondiabetic counterparts. The proportion of patients with LDL-C concentrations requiring lifestyle (>2.6 mmol/L [>100 mg/dL]) or drug (>3.4 mmol/L [>130 mg/dL]) intervention decreased from 78% and 42% to 66% and 26%, respectively. CONCLUSIONS: Low HDL-C is the most frequent dyslipidemic disorder in patients with poorly controlled insulin-treated type 1 diabetes, and a high proportion show LDL-C levels requiring intervention. Less favorable lipid profiles could explain the absence of sex protection in diabetic women. The improvement caused by glycemic optimization puts forward intensive therapy as the initial treatment of choice for dyslipidemia in poorly controlled type 1 diabetes. PMID- 11025786 TI - Pulse pressure and mortality in older people. AB - BACKGROUND: In older people, observational data are unclear concerning the relationships of systolic and diastolic blood pressure with cardiovascular and total mortality. We examined which combinations of systolic, diastolic, pulse, and mean arterial pressure best predict total and cardiovascular mortality in older adults. METHODS: In 1981, the National Institute on Aging initiated its population-based Established Populations for Epidemiologic Studies of the Elderly in 3 communities. At baseline, 9431 participants, aged 65 to 102 years, had blood pressure measurements, along with measures of medical history, use of medications, disability, and physical function. During an average follow-up of 10. 6 years among survivors, 4528 participants died, 2304 of cardiovascular causes. RESULTS: In age- and sex-adjusted survival analyses, the lowest overall death rate occurred among those with systolic pressure less than 130 mm Hg and diastolic pressure 80 to 89 mm Hg; relative to this group, the highest death rate occurred in those with systolic pressure of 160 mm Hg or more and diastolic pressure less than 70 mm Hg (relative risk, 1.90; 95% confidence interval, 1.47 2.46). Both low diastolic pressure and elevated systolic pressure independently predicted increases in cardiovascular (P<.001) and total (P<.001) mortality. Pulse pressure correlated strongly with systolic pressure (R = 0.82) but was a slightly stronger predictor of both cardiovascular and total mortality. In a model containing pulse pressure and other potentially confounding variables, diastolic pressure (P =.88) and mean arterial pressure (P =.11) had no significant association with mortality. CONCLUSIONS: Pulse pressure appears to be the best single measure of blood pressure in predicting mortality in older people and helps explain apparently discrepant results for low diastolic blood pressure. PMID- 11025788 TI - Neurologic manifestations of infective endocarditis: a 17-year experience in a teaching hospital in Finland. AB - BACKGROUND: Many previous studies have endeavored to find appropriate means to reduce the occurrence of neurologic manifestations in patients with infective endocarditis (IE). We evaluated patients with IE-associated neurologic complications and compared them with patients with IE who did not have neurologic symptoms. Particular attention was focused on assessing the impact of cardiac surgery and the presence of potential risk factors for complications on the outcome of the patients. METHODS: A total of 218 episodes designated as definite or possible IE according to Duke criteria and treated during the years 1980 through 1996 in a Finnish teaching hospital were retrospectively evaluated for neurologic manifestations. RESULTS: Neurologic complications were identified in 55 episodes (25%), with an embolic event as the most frequent manifestation (23/55; 42%). In the majority (76%) of episodes, the neurologic manifestation was evident before antimicrobial treatment was started, being the first sign of IE in 47% of episodes. Only 1 recurrent cerebral embolization was observed. Neurologic complications were significantly associated with Staphylococcus aureus infection (29% vs 10%; P =.001) and with IE affecting both the aortic and the mitral valves (56% vs 23%; P<.01), but not with echocardiographic detection of vegetations or anticoagulant therapy. Death during the acute phase of IE occurred in 13 episodes (24%) with neurologic complications and in 17 episodes (10%) without neurologic complications (P<.03). In episodes with neurologic complications, the IE associated mortality rate was 25% (10/40) in the medical treatment group and 20% (3/15) in the surgical group. No neurologic deterioration was observed in these surgically treated patients postoperatively. CONCLUSIONS: Our results reinforce the belief that rapid diagnosis and initiation of antimicrobial therapy may still be the most effective means to prevent neurologic complications. These data underscore the importance of diagnostic alertness to the prognosis of patients with IE. PMID- 11025787 TI - Cost-effectiveness of new antiplatelet regimens used as secondary prevention of stroke or transient ischemic attack. AB - BACKGROUND: Compared with aspirin alone, use of the new antiplatelet regimens, including aspirin combined with dipyridamole and clopidogrel bisulfate, has been found to further reduce the risk of stroke and other vascular events in patients who have experienced stroke or transient ischemic attack. However, their cost effectiveness ratios relative to aspirin alone have not been estimated. METHODS: We developed a Markov model to measure the clinical benefits and the economic consequences of the following strategies to treat high-risk patients aged 65 years or older: (1) aspirin, 325 mg/d; (2) aspirin, 50 mg/d, and dipyridamole, 400 mg/d; and (3) clopidogrel bisulfate, 75 mg/d. Input data were obtained by literature review. Outcomes were expressed as US dollars per quality-adjusted life-year (QALY). RESULTS: The use of aspirin combined with dipyridamole was more effective and less costly compared with the use of aspirin alone, providing a gain of 0.3 QALY for a 65-year-old patient. This regimen remained cost-effective despite wide sensitivity analyses. Clopidogrel was more effective and more costly compared with aspirin alone, yielding a gain of 0.2 QALY with a marginal cost effectiveness ratio of $26,580 per each additional QALY (patient aged 65 years). Sensitivity analyses demonstrated that the efficacy of clopidogrel and its cost were key factors in determining its cost-effectiveness ratio compared with aspirin, which exceeded $50,000 when its efficacy decreased by half or its cost doubled. CONCLUSION: To prevent stroke in high-risk patients, dipyridamole combined with aspirin was more effective and less costly than aspirin alone, and clopidogrel was cost-effective compared with current standards of medical practice, except in extreme scenarios. PMID- 11025789 TI - Temporal association between hospitalization and rate of falls after discharge. AB - BACKGROUND: Evidence suggests that acute illness and hospitalization may increase the risk for falls. OBJECTIVE: To evaluate the rate of falls, and associated risk factors, for 90 days following hospital discharge. METHODS: We consecutively enrolled 311 patients, aged 65 years and older, discharged from the hospital after an acute medical illness and receiving home-nursing services. Patients were assessed within 5 days of discharge for prehospital and current functioning by self-report, and balance, vision, cognition, and delirium by objective measures. Patients were followed up weekly for 13 weeks for falls, injuries, and health care use. RESULTS: The rate of falls was significantly higher in the first 2 weeks after hospitalization (8.0 per 1000 person-days) compared with 3 months later (1.7 per 1000 person-days) (P =.002). Fall-related injuries accounted for 15% of all hospitalizations in the first month after discharge. Independent prehospital risk factors significantly associated with falls included dependency in activities of daily living, use of a standard walker, 2 or more falls, and more hospitalizations in the year prior. Posthospital risk factors included use of a tertiary amine tricyclic antidepressant, probable delirium, and poorer balance, while use of a cane was protective. CONCLUSIONS: The rate of falls is substantially increased in the first month after medical hospitalization, and is an important cause of injury and morbidity. Posthospital risk factors may be potentially modifiable. Efforts to assess and modify risk factors should be integral to the hospital and posthospital care of older adults (those aged >/=65 years). PMID- 11025790 TI - Physicians' attitudes about prescribing and knowledge of the costs of common medications. AB - BACKGROUND: Compliance with medical therapy is often compromised because patients cannot afford to pay for medications. Inadequate physician knowledge of drug costs may unwittingly contribute to this problem. OBJECTIVE: To measure attitudes about prescribing and knowledge of medication costs and compare differences among attending physicians and residents. DESIGN/PARTICIPANTS: Written survey of internal medicine house staff and general medicine attending physicians in an urban hospital-based primary care center. RESULTS: One hundred thirty-four of 189 physicians responded (71% response rate). Seventy percent of respondents were house officers and 30% were attending physicians. Eighty-eight percent of physicians felt the cost of medicines was an important consideration in the prescribing decision, and 71% were willing to sacrifice some degree of efficacy to make drugs more affordable for their patients. However, 80% often felt unaware of the actual costs. Only 33% had easy access to drug cost data, and only 13% had been formally educated about drug costs. Regarding insurance coverage, 94% of physicians gave strong consideration to the cost of medications when patients were self-paying, 68% when patients had Medicare, and 30% when patients had Medicaid or were participants in a health maintenance organization with a prescription plan. Physicians' estimates of the cost of a month's supply of 33 commonly used medications were accurate in 45% of cases, too low for 40%, and too high for 15%. The costs of brand-name and expensive drugs were most likely to be underestimated. House officers were less cost-conscious than attending physicians. CONCLUSIONS: Physicians were predisposed to being cost-conscious in their prescribing habits, but lacked accurate knowledge about actual costs and insurance coverage of drugs. Interventions are needed to educate physicians about drug costs and provide them with reliable, easily accessible cost information in real-world practice. PMID- 11025791 TI - Gastrointestinal tract symptoms among persons with diabetes mellitus in the community. AB - BACKGROUND: Gastrointestinal (GI) tract symptoms are common among patients with diabetes mellitus (DM) seen in tertiary care centers. The degree to which this reflects referral bias is unclear. OBJECTIVES: To determine whether GI tract symptoms are more prevalent in unselected patients with DM from the general community compared with their age- and sex-matched counterparts without DM and to assess the association of GI tract symptoms in persons with DM with psychosomatic symptoms, medication use, and symptoms of autonomic neuropathy. METHODS: In this population-based, cross-sectional study, Olmsted County, Minnesota, residents with type 1 DM, a random sample of residents with type 2 DM, and 2 age- and sex stratified random samples of nondiabetic residents (total of 1262 person for the 4 groups) were mailed a previously validated symptom questionnaire. RESULTS: Heartburn was less common in residents with type 1 DM vs controls (12% vs 23%; P<.05). No significant difference in prevalence was detected (residents with type 1 DM vs controls; residents with type 2 DM vs controls) for nausea or vomiting (12% vs 11%; 6% vs 6%), dyspepsia (19% vs 21%; 13% vs 17%), or constipation (17% vs 14%; 10% vs 12%). However, constipation and/or laxative use was slightly more common in residents with type 1 DM (27% vs 19%; P<.15), particularly in men, and was associated with the intake of calcium channel blockers. CONCLUSIONS: In the community, the prevalence of most GI tract symptoms is similar in persons with or without DM, except for a lower prevalence of heartburn and an increased prevalence of constipation or laxative use in residents with type 1 DM, especially in men. This difference is associated with calcium channel blocker use rather than symptoms of autonomic neuropathy. In community-based practices, physicians should not immediately assume that GI tract symptoms in patients with DM represent a complication of DM. PMID- 11025792 TI - The incidence of antimicrobial allergies in hospitalized patients: implications regarding prescribing patterns and emerging bacterial resistance. AB - BACKGROUND: The development of antimicrobial guidelines is one way in which institutions attempt to control emerging resistance, but the real challenge falls on promoting and ensuring adherence to these guidelines. Investigating reasons for the prescribing of alternative antimicrobial agents outside of these guidelines is crucial for modifying practices that may adversely impact institutional antimicrobial goals. METHODS: Retrospective cross-referencing of computerized pharmacy printouts and concurrent manual medical record review. RESULTS: Approximately 25% (470/1893) of the patients requiring antimicrobial therapy reported an allergy to at least 1 antimicrobial agent. The most commonly reported antimicrobial allergy was penicillin (295/1893 [15.6%]). Eighty-five patients (18.1%) reported having an allergy to 2 or more antimicrobial agents. Only 4% (27/601) of the reported antimicrobial allergies contained documentation as to the nature of the specific allergic reactions, while a manual medical record review revealed that 32% (23/73) of the antimicrobial allergies contained documentation of the specific allergic reaction. Ninety-eight (39. 7%) of 247 patients reporting an allergy only to penicillin and/or cephalosporin received vancomycin in comparison with 247 (17.4%) of 1423 patients without any antimicrobial allergies (P<.001). Similarly, 53 (21.5%) of 247 patients with reported penicillin and/or cephalosporin allergies received levofloxacin compared with 114 (8.0%) of 1423 patients without any antimicrobial allergy (P<. 001). CONCLUSION: The incidence of penicillin allergy at our institution exceeds population averages. This finding, in combination with limited documentation of drug allergies, appears to lead to the prescribing of alternative antimicrobial agents that do not fit into institutional antimicrobial guidelines and, in some instances, may put the patient at risk for infection and/or colonization with resistant organisms. Use of these alternative agents may adversely impact the ability to manage emerging antimicrobial resistance. PMID- 11025793 TI - Prescribing potentially inappropriate psychotropic medications to the ambulatory elderly. AB - BACKGROUND: Psychotropic agents account for 23% to 51% of all inappropriate medications prescribed based on 1991 inappropriate medication criteria for nursing home residents. The criteria were revised to apply to all people older than 65 years. This study used the revised criteria in ambulatory settings to quantify potentially inappropriate prescription of psychotropic agents and identify associated characteristics. METHODS: The 1996 public use data files of the National Ambulatory Medical Care Survey and the National Hospital Ambulatory Medical Care Survey were analyzed for inappropriate prescription of psychotropic medications for the elderly in office-based settings and outpatient departments. Disease-independent and disease-dependent criteria were analyzed. RESULTS: Elderly patients were prescribed a psychotropic agent in 8. 7% of all visits, antidepressant and antianxiety agents being the most frequently prescribed medications. Commonly, elderly patients receiving psychotropic agents were female, white, aged between 65 and 74 years, and received health care in a metropolitan area. Potentially inappropriate psychotropic agents were prescribed in 27. 2% of all visits involving a psychotropic agent for the elderly. Disease independent criteria (eg, antidepressant agents and long-acting benzodiazepines) accounted for most of the potentially inappropriate prescriptions. Factors positively associated with potentially inappropriate prescriptions included older age, "seen before" status, and antidepressant drug class, while enrollment in Medicaid, antipsychotic drug class, living in the Northeast region, and receiving health care in a metropolitan area were negatively associated. CONCLUSIONS: Potentially inappropriate prescription of psychotropic agents is very common for the elderly patient in the ambulatory setting. By focusing on the agents most frequently involved (eg, amitriptyline and long-acting benzodiazepines), provider characteristics (eg, location), and patient characteristics (eg, age), the greatest impact on potentially inappropriate prescribing can be achieved. PMID- 11025795 TI - Impact of body weight on blood pressure with a focus on sex differences: the Tromso Study, 1986-1995. AB - BACKGROUND: The prevalence of obesity and hypertension is increasing in Western societies. We examined the effects of initial body mass index ([BMI] weight in kilograms divided by height in meters squared) and change in BMI on change in blood pressure, and we assessed sex differences. METHODS: A general population in the municipality of Tromso, northern Norway, was examined in 1986 and 1987 and again in 1994 and 1995. Altogether, 75% of the individuals, women aged 20 to 56 years and men aged 20 to 61 years, attended the baseline examination. A total of 15,624 individuals (87% of all still living in the municipality) were examined twice. RESULTS: Mean BMI increased between the examinations, more for the younger than the older examinees, and also more among women than men (P<.001). Adjusted for several covariates, BMI change was associated with systolic and diastolic blood pressure change for both sexes (regression coefficients: 1.43 [95% confidence interval (CI), 1.23-1. 64] and 0.90 [95% CI, 0.76-1.04], respectively, for men; and 1.24 [95% CI, 1.09-1.39] and 0.74 [95% CI, 0.63-0.84] for women). Baseline BMI was associated with systolic and diastolic blood pressure change for women only (regression coefficients: 0.38 [95% CI, 0.30-0.47] and 0.17 [95% CI, 0.11-0.23], respectively). CONCLUSIONS: For women, both BMI at baseline and BMI change were independently associated with blood pressure change. For a given increase in BMI, obese women had a greater increase in blood pressure than lean women. This was not the case for men, for whom BMI change was the only significant predictor. Furthermore, a BMI increase for obese women induced a greater systolic blood pressure increase compared with men. PMID- 11025794 TI - The Gaucher registry: demographics and disease characteristics of 1698 patients with Gaucher disease. AB - BACKGROUND: The Gaucher Registry, the largest database of patients with Gaucher disease (GD) worldwide, was initiated to better delineate the progressive nature of the disorder and determine optimal therapy. This report describes the demographic and clinical characteristics of 1698 patients with GD before they received enzyme replacement therapy. METHODS: Physicians worldwide who treat patients with GD were invited to submit prospective and retrospective data for an ongoing registry, using standardized data collection forms, for central processing and review. RESULTS: Most patients were from the United States (45%) and Israel (17%), but patients are from 38 countries. Most (94%) had type 1 GD, fewer than 1% had type 2, and 5% had type 3. Mutant allele frequency data, available for 45% of patients, showed the most common alleles to be N370S (53%), L444P (18%), 84GG (7%), and IVS2+1 (2%). Twenty-five percent of L444P homozygotes (13 of 52 patients) had type 1 GD phenotype. Mean age at diagnosis in patients with the N370S/N370S genotype was 27.2 years (SD, 19.7 years); in L444P/L444P patients, 2. 3 years (SD, 3.2 years). Histories of bone pain and radiological bone disease were reported by 63% and 94% of patients, respectively; both were more likely in asplenic patients than in patients with spleens. Mean spleen and liver volumes were 19.8 and 2.0 multiples of normal, respectively. Anemia and thrombocytopenia were present in 64% and 56%, respectively. Thrombocytopenia was present in 13% of asplenic patients. CONCLUSIONS: The Gaucher Registry permits a comprehensive understanding of the clinical spectrum of GD because of the uniquely large sample size. The Registry will be useful in evaluating the effects of specific therapies in GD and the possible influences of environment, ethnicity, and genotype on the natural history of the disorder. PMID- 11025796 TI - Systematic review of clinical prediction rules for neuroimaging in the evaluation of dementia. AB - BACKGROUND: Clinical practice guidelines for dementia do not recommend routine neuroimaging but vary in their recommended clinical prediction rules to identify patients who should undergo neuroimaging for potentially reversible causes of dementia. METHODS: Using a MEDLINE search supplemented by other strategies, we identified studies from January 1, 1983, through December 31, 1998, that evaluated the diagnostic performance of a clinical prediction rule. We calculated the sensitivity and specificity of each rule, then evaluated their diagnostic performance in a hypothetical cohort of 1000 patients with dementia, varying the prevalence of potentially reversible dementia from 1% to 15%. RESULTS: We identified 7 studies that evaluated at least 1 of 6 different clinical prediction rules. Only one rule consistently had high sensitivity (>85%) across all studies; none consistently had high specificity (>85%). Six of the 7 studies included less than 15 cases of potentially reversible dementia; thus the sensitivity and specificity for each rule had relatively wide confidence intervals. At a 5% prevalence of potentially reversible dementia, all rules had low positive predictive value (<15%) in our hypothetical cohort. Depending on the rule, our analysis predicts 6 to 44 of the 50 patients with potentially reversible dementia (5% prevalence in cohort of 1000 patients) would not undergo imaging. CONCLUSIONS: There is considerable uncertainty in the evidence underlying clinical prediction rules to identify which patients with dementia should undergo neuroimaging. Application of these rules may miss patients with potentially reversible causes of dementia. PMID- 11025797 TI - Lack of positive correlation between statin use and bone mineral density in Japanese subjects with type 2 diabetes. PMID- 11025799 TI - Paraneoplastic limbic encephalitis should not be overlooked as a possible cause of delirium in cancer patients. PMID- 11025798 TI - Home, ambulatory, and office blood pressure measurements. PMID- 11025800 TI - Delirium as a predictor of survival in older patients with advanced cancer. PMID- 11025804 TI - A piece of my mind: hands that held other hands. PMID- 11025802 TI - Cranial computed tomography before lumbar puncture. PMID- 11025805 TI - JAMA 100 years ago: MUNICIPAL REGULATION OF THE SPITTING HABIT PMID- 11025806 TI - Microvascular free-flap reconstruction in the head and neck. PMID- 11025807 TI - Green light for federally funded research on embryonic stem cells. PMID- 11025808 TI - ABIM Foundation launches forum 2000: framing a quality agenda for health care. American Board of Internal Medicine. PMID- 11025811 TI - The world in medicine: No chimp DNA in vaccine PMID- 11025809 TI - Panel predicts shortfall in care for the aged. PMID- 11025812 TI - The world in medicine: recessive alzheimer gene? PMID- 11025813 TI - The world in medicine: HIV misdiagnoses PMID- 11025814 TI - The world in medicine: global alcohol risks PMID- 11025815 TI - Rehabilitation for traumatic brain injury. PMID- 11025816 TI - Rehabilitation for traumatic brain injury. PMID- 11025817 TI - Rehabilitation for traumatic brain injury PMID- 11025818 TI - Exercise intensity and risk of chronic disease. PMID- 11025819 TI - Exercise intensity and risk of chronic disease PMID- 11025820 TI - Quality of evidence for treatment of prostate cancer. PMID- 11025821 TI - Quality of evidence for treatment of prostate cancer PMID- 11025822 TI - Should patients in quality-improvement activities have the same protections as participants in research studies? PMID- 11025823 TI - Should patients in quality-improvement activities have the same protections as participants in research studies? PMID- 11025824 TI - Should patients in quality-improvement activities have the same protections as participants in research studies? PMID- 11025825 TI - Should patients in quality-improvement activities have the same protections as participants in research studies? PMID- 11025826 TI - Should patients in quality-improvement activities have the same protections as participants in research studies? PMID- 11025827 TI - Would prevention of gun carrying reduce US homicide rates? PMID- 11025828 TI - Would prevention of gun carrying reduce US homicide rates? PMID- 11025829 TI - Would prevention of gun carrying reduce US homicide rates? PMID- 11025830 TI - Would prevention of gun carrying reduce US homicide rates? PMID- 11025831 TI - Risk of breast cancer with oral contraceptive use in women with a family history of breast cancer. AB - CONTEXT: Oral contraceptive (OC) use is weakly associated with breast cancer risk in the general population, but the association among women with a familial predisposition to breast cancer is less clear. OBJECTIVE: To determine whether the association between OC use and risk of breast cancer is influenced by family history of the disease. DESIGN AND SETTING: Historical cohort study of 426 families of breast cancer probands diagnosed between 1944 and 1952 at the Tumor Clinic of the University of Minnesota Hospital. Follow-up data on families were collected by telephone interview between 1991 and 1996. PARTICIPANTS: A total of 394 sisters and daughters of the probands, 3002 granddaughters and nieces, and 2754 women who married into the families. MAIN OUTCOME MEASURE: Relative risk (RR) of breast cancer associated with history of OC use by relationship to proband. RESULTS: After accounting for age and birth cohort, ever having used OCs was associated with significantly increased risk of breast cancer among sisters and daughters of the probands (RR, 3.3; 95% confidence interval [CI], 1.6-6.7), but not among granddaughters and nieces of the probands (RR, 1.2; 95% CI, 0.8 2.0) or among marry-ins (RR, 1.2; 95% CI, 0.8-1.9). Results were essentially unchanged after adjustment for parity, age at first birth, age at menarche, age at menopause, oophorectomy, smoking, and education. The elevated risk among women with a first-degree family history of breast cancer was most evident for OC use during or prior to 1975, when formulations were likely to contain higher dosages of estrogen and progestins (RR, 3.3; 95% CI, 1.5-7.2). A small number of breast cancer cases (n = 2) limited the statistical power to detect risk among women with a first-degree relative with breast cancer and OC use after 1975. CONCLUSIONS: These results suggest that women who have ever used earlier formulations of OCs and who also have a first-degree relative with breast cancer may be at particularly high risk for breast cancer. Further studies of women with a strong family history who have used more recent lower-dosage formulations of OCs are needed to determine how women with a familial predisposition to breast cancer should be advised regarding OC use today. JAMA. 2000;284:1791-1798. PMID- 11025832 TI - Differences in prognostic factors and outcomes between women and men undergoing coronary artery stenting. AB - CONTEXT: Women with coronary artery disease (CAD) are believed to have a higher risk for adverse outcomes than men after conventional coronary interventions. The increasing use of coronary stenting has improved the outcome of patients undergoing coronary interventions, but little is known about the nature of outcomes in men vs women after this procedure. OBJECTIVE: To examine whether there are sex-based differences in prognostic factors and in early and late outcomes among CAD patients undergoing coronary stent placement. DESIGN, SETTING, AND PATIENTS: Inception cohort study, at 2 tertiary referral institutions in Germany. Consecutive series of 1001 women and 3263 men with symptomatic CAD who were treated with stenting between May 1992 and December 1998. Patients who underwent stenting in the setting of acute myocardial infarction were excluded. MAIN OUTCOME MEASURE: The combined event rates of death and nonfatal myocardial infarction, assessed at 30 days and 1 year after stenting and compared by sex. RESULTS: Compared with men, women undergoing coronary stenting were significantly older (mean age, 69 vs 63 years) and more likely to present with diabetes, arterial hypertension, or hypercholesterolemia. Women had less extensive CAD, a less frequent history of myocardial infarction and better preserved left ventricular function than men. Women presented an excess risk of death or nonfatal myocardial infarction only during the early period after stenting: the 30-day combined event rate of death or myocardial infarction was 3.1% in women and 1.8% in men (P =.02) and the multivariate-adjusted hazard ratio (HR) for women was 2.02 (95% confidence interval [CI], 1.27-3.19). At 1 year, the outcome was similar for both women and men (combined event rate for women, 6.0%, and for men, 5.8% (P =.77); multivariate-adjusted HR for women, 1.06 [95% CI, 0.75 1.48]). There was a sex difference in the prognostic value of baseline characteristics: the strongest prognostic factors were diabetes in women and age in men. CONCLUSIONS: The results of this study indicate that 1-year outcomes of women with CAD undergoing coronary artery stenting are similar to those of men. Despite the similarity in outcomes, there are several sex-specific differences in baseline characteristics, clinical course after the intervention, and relative weight of prognostic factors. JAMA. 2000;284:1799-1805. PMID- 11025833 TI - Efficacy of 3 commonly used hearing aid circuits: A crossover trial. NIDCD/VA Hearing Aid Clinical Trial Group. AB - CONTEXT: Numerous studies have demonstrated that hearing aids provide significant benefit for a wide range of sensorineural hearing loss, but no carefully controlled, multicenter clinical trials comparing hearing aid efficacy have been conducted. OBJECTIVE: To compare the benefits provided to patients with sensorineural hearing loss by 3 commonly used hearing aid circuits. DESIGN: Double-blind, 3-period, 3-treatment crossover trial conducted from May 1996 to February 1998. SETTING: Eight audiology laboratories at Department of Veterans Affairs medical centers across the United States. PATIENTS: A sample of 360 patients with bilateral sensorineural hearing loss (mean age, 67.2 years; 57% male; 78.6% white). INTERVENTION: Patients were randomly assigned to 1 of 6 sequences of linear peak clipper (PC), compression limiter (CL), and wide dynamic range compressor (WDRC) hearing aid circuits. All patients wore each of the 3 hearing aids, which were installed in identical casements, for 3 months. MAIN OUTCOME MEASURES: Results of tests of speech recognition, sound quality, and subjective hearing aid benefit, administered at baseline and after each 3-month intervention with and without a hearing aid. At the end of the experiment, patients ranked the 3 hearing aid circuits. RESULTS: Each circuit markedly improved speech recognition, with greater improvement observed for soft and conversationally loud speech (all 52-dB and 62-dB conditions, P 15 IU/L, n = 11). As expected, late perimenopausal women had irregular and longer menstrual cycles, lower estradiol (E(2)) and estrone (E(1)) levels, and a lower frequency of ovulations vs. the early group. ALP was higher (76.5 +/- 8.3 vs. 58.3 +/- 2.7 IU/L, p = 0.045) compared with the early perimenopausal group. In a subsample (n = 10), OC was associated with ALP (r = 0.69, p < 0.03), FSH was positively related to ucOC concentrations (r = 0.7, p < 0.03), and women with E(1) concentrations <40 pg/ml had double the percentage of ucOC compared with those where E(1) was >40 pg/ml (46.3% +/- 6.6% vs. 22.0% +/- 3. 1%, p < 0.006). Clinical markers of the perimenopause are associated with a nonspecific but inexpensive marker of enhanced bone turnover (i.e., higher ALP) and correlate well with more precise markers of bone activity. These findings suggest that health-promotion strategies for preserving bone should be instituted well before the last menstrual period. PMID- 11025866 TI - Optimizing percutaneous coronary revascularization in diabetic women: analysis from the EPISTENT trial. AB - Clinical studies have demonstrated that diabetic women have an increased risk of death and nonfatal myocardial infarction (MI) after percutaneous coronary intervention (PCI). However, there have been few data regarding the outcome of diabetic women in the current era of percutaneous coronary revascularization. Using the Evaluation of Platelet IIb/IIIa Inhibitor for Stenting Trial (EPISTENT) database, we determined morbidity and mortality for diabetic women undergoing PCT in the current era using stents and the glycoprotein IIb/IIIa inhibitor, abciximab. There were no mortality differences in the diabetic women treated with stent-placebo, stent-abciximab, or balloon-abciximab at 30 days and 6 months. However, the primary end point of 1-year death, MI, or target vessel revascularization (TVR) was lowered in the diabetic women from 34.5% in the stent placebo group and 28.9% in the balloon-abciximab group to 13. 3% in the stent abciximab group (p = 0.02 for stent-stent comparison, and p = 0.09 for stent abciximab vs. balloon-abciximab comparison). Also, 1-year TVR rates were lowered from 21.1% in the stent-placebo group and 26.7% in the balloon-abciximab group to 4.5% in the stent-abciximab group (p = 0.02 for stent-stent comparison, and p = 0.004 for stent-abciximab vs. balloon-abciximab comparison). The combination of stenting and abciximab therapy among diabetic women resulted in a significant reduction in 1-year rate of death, MI, or TVR compared with stent-placebo or balloon-abciximab therapy. PMID- 11025867 TI - Prevalence and clustering of menopausal symptoms in older women by hysterectomy and oophorectomy status. AB - This study examines the association of hysterectomy and oophorectomy with the prevalence and clustering of menopausal symptoms in a large population-based sample of older women. Subjects were 1121 women aged 50-89 from the Rancho Bernardo Study. Information on menopause, hysterectomy, oophorectomy, estrogen use, and other covariates was obtained in 1984-1987. A 1989 mailed survey obtained information on menopausal symptoms. In this sample, 22.1% reported hysterectomy with bilateral oophorectomy, and 25.3% reported hysterectomy with ovarian conservation. Mean time since hysterectomy was 26 (+/-12) years. Overall, 37% reported current estrogen use, and 40% reported past use. The duration of estrogen use was longer for women who had a hysterectomy (p < 0.001). Age adjusted comparisons indicated that more women who had a hysterectomy, with or without bilateral oophorectomy, reported greater energy after menopause (p = 0.003 and p = 0.001, respectively), and more women with bilateral oophorectomy reported greater interest in sex (p = 0.007) and that life was getting better (p = 0.012) than women with natural menopause. Principal components factor analysis of the symptom data for all women yielded four factors: psychological, vasomotor, positive feelings, and self-image. Analyses performed within each group of women yielded similar factors and loadings. Adjusted comparisons of factor scores indicated that positive feelings were significantly higher in women who had a hysterectomy, with or without bilateral oophorectomy (p < 0.01) than in women with natural menopause. This difference was limited to current estrogen users. Vasomotor symptoms, psychological symptoms, and negative self-image did not differ by hysterectomy or oophorectomy status before or after stratification for estrogen use (p > 0.10). This study found after a hysterectomy, women are more likely to recall positive feelings about their menopause than women with natural menopause. Relief from symptoms leading to hysterectomy and use of replacement estrogen may be partly responsible. Results do not support the thesis that surgical menopause is associated with a sustained increased prevalence of vasomotor, psychological, or other symptoms. PMID- 11025868 TI - Premenstrual symptom severity: impact on social functioning and treatment-seeking behaviors. AB - Our purpose was to understand premenstrual symptomatology and treatment-seeking behaviors by examining three subjective measurement approaches for premenstrual syndrome (PMS), their relationship to social functioning interference, and the role of symptom severity in a broader model of help seeking for PMS. Cross sectional data were obtained from 1022 mail survey respondents who were derived from a nationally representative random sample of women, aged 18-49, experiencing regular menstrual cycles. Statistical analyses included Pearson correlations, chi square tests, t tests, and logistic regression. The three symptom severity measures (a global self-appraisal, summative symptom counting, and categorical configural) were strongly intercorrelated, ranging from. 60 to.78 (p < 0.001), and were correlated with interference in social and occupational functioning domains, ranging from.44 to.77 (all p < 0.001). A global self-report measure identified 4.9% of women with severe symptoms, whereas a DSM-IV-adapted approach identified 11.3% with premenstrual dysphoric disorder (PMDD) symptoms (of whom 92% also reported social interference). PMS treatment seeking was predicted by older age, PMS symptoms experienced in most cycles, greater self-reported symptom severity, greater overall use of healthcare services, and less negative attitudes toward PMS (all p < 0.05). These findings support the feasibility of clinician's use of brief screening approaches for PMDD, especially using short summative symptom rating scales. Women underidentify the severity of their PMS difficulties despite the reported difficulties associated with consistent social and occupational interference in most life domains. They are also reluctant to seek help for treatable PMS symptoms because of attitudinal barriers regardless of the severity of their PMS symptoms. PMID- 11025869 TI - Physical activity and functional capacity measurement in women: a report from the NHLBI-sponsored WISE study. AB - Physical activity and functional capacity have not been assessed by questionnaire for criterion validity in women. We wished to evaluate the ability of a physical activity and a functional capacity assessment questionnaire to predict functional capacity measured by treadmill exercise stress testing, as well as correlate with cardiac risk factors and angiographic coronary artery disease (CAD) in women. In a National Heart, Lung and Blood Institute (NHLBI)-sponsored cross-sectional population study involving four academic medical centers, 476 women with cardiac risk factors undergoing coronary angiography for evaluation for suspected myocardial ischemia were enrolled in the Women's Ischemia Syndrome Evaluation (WISE). The main outcome measures were functional capacity measured during symptom-limited exercise treadmill testing, cardiac risk factors, and CAD, using core laboratory-determined measures. Physical activity measured by the Postmenopausal Estrogen and Progesterone Intervention physical activity questionnaire (PEPI-Q) and functional capacity measured by the Duke Activity Status Index (DASI) questionnaire, correlated with functional capacity measured in metabolic equivalents (METS), as estimated during symptom-limited exercise treadmill testing (r = 0.27, p = 0.001 and r = 0.31, p = 0. 0002, respectively). The DASI was a significant independent predictor of functional capacity even after adjustment for cardiac risk factors, and the PEPI-Q was not. The DASI and PEPI-Q scores were inversely associated with higher numbers and levels of cardiac risk factors, as well as angiographic CAD. The DASI questionnaire is a reasonable correlate of functional capacity achieved during symptom-limited treadmill exercise testing in women with suspected myocardial ischemia. Lower functional capacity or physical activity measured by the DASI and PEPI-Q, respectively, is associated with more prevalent cardiac risk factors and angiographic CAD. These findings suggest that the DASI and, to a lesser extent, the PEPI-Q have criterion validity for use in health-related research in women. PMID- 11025870 TI - Improving follow-up after an abnormal pap smear: results from a quasi experimental intervention study. AB - The success of cervical cancer control programs depends on regular screening with the Pap smear test and prompt and appropriate treatment of early neoplastic lesions. Recognizing the potentially grave consequences of lack of follow-up for abnormal Pap smears, numerous intervention studies have tested the impact of a variety of strategies to increase return for follow-up. The majority of these studies were evaluated under controlled experimental conditions. Despite the encouraging findings of these trials, the next step in the research continuum requires that the effectiveness of these interventions be demonstrated in real world settings before full implementation is initiated. We report the results of an evaluation study assessing the combined effectiveness of three intervention modalities found effective in prior randomized studies: a tracking follow-up protocol, transportation incentives, and financial incentives. This study used a before-after, nonequivalent control group design to assess the impact of a multifaceted intervention that included a computerized tracking protocol with transportation and financial incentives. The study was implemented at two major hospitals, two comprehensive health centers (CHC), and nine public health centers (PHC) under the jurisdiction of the Los Angeles County Department of Health Services. One hospital, one CHC, and the four PHC located in the catchment area of the CHC were selected as experimental sites. The control sites - one hospital, one CHC, and five PHC - provided usual care. All women with an abnormal Pap smear at the intervention and control sites were included in the study. The study consisted of a 1-year period of baseline data collection (September 1989-August 1990), followed by a 2(1/2)-year intervention period (September 1990-February 1993). During the intervention period, the intervention protocol was implemented at the experimental sites, and the control sites provided usual care. Overall, we found that the rates of receipt of follow-up care were consistent with those found in similar studies. In contrast to results obtained in these prior randomized trials, we did not find strong and consistent evidence for intervention effects. Significant findings emerged only at the CHC and hospital levels and only for selected years. Results underscore the importance of testing interventions in real world conditions before large-scale implementation is initiated. In addition, this study highlights the challenge of detecting intervention effects in large-scale studies because of the greater measurement difficulties in field studies as compared with controlled experiments. PMID- 11025871 TI - U.S. Women Physicians' personal and clinical breast cancer screening practices. AB - Little is known about predictors of physicians' personal or clinical compliance with breast cancer screening recommendations. We explored this in 4501 respondents to the Women Physicians' Health Study, a questionnaire-based study of a representative sample of U.S. women M. D.s. Overall, 21% of women physicians performed breast self-examination (BSE) at least monthly, about two thirds had received a clinical breast examination (CBE) within the last year, and 85% had received one within the last 2 years. Of those <40 years old, 14% had received a mammogram in the past year, as had 42% of those 40-49 and 59% of those 50-70 years old. Being a primary care practitioners or obstetrician/gynecologist was a significant predictor of counseling or screening for CBE and mammography. Only 46% of all women physicians reported discussing or performing mammograms at least once a year for those >/=50- 3.0 and the amplitude of the TWA was > 1.8 microV. Twelve (20%) of the included 60 patients showed a positive TWA. The sensitivity concerning a previous arrhythmic event amounted to 65%, the specificity up to 98%, respectively. The alternans ratio was significantly higher in patients with a previous event (30.3 +/- 53.2 vs 2.9 +/- 5.9, P < 0.001) and cumulative alternans voltage (4.67 +/- 3.55 vs 1.75 +/- 1.88 microV, P < 0.001). In 19 patients, invasively investigated by an electrophysiological study, a significant correlation between inducibility of tachyarrhythmias and a positive TWA result was found (Spearman R = 0.51, P = 0.01). In conclusion, the TWA analysis seems to identify patients with nonischemic cardiomyopathy who are at an increased risk of ventricular tachyarrhythmias. PMID- 11025896 TI - Dispersion of QT intervals: a measure of dispersion of repolarization or simply a projection effect? AB - QT interval dispersion may provide little information about repolarization dispersion. Some clinical measurements demonstrate an association between high QT interval dispersion and high morbidity and mortality, but what is being measured is not clear. This study was designed to help resolve this dilemma. We compared the association between different clinical measures of QT interval dispersion and the ECG lead amplitudes derived from a heart vector model of repolarization with no repolarization dispersion whatsoever. We compared our clinical QT interval dispersion data obtained from 25 subjects without cardiac disease with similar data from published studies, and correlated these QT dispersion results with the distribution of lead amplitudes derived from the projection of the heart vector onto the body surface during repolarization. Published results were available for mean relative QT intervals and mean differences from the maximum QT interval. The leads were derived from Uijen and Dower lead vector data. Using the Uijen lead vector data, the correlation between measurements of dispersion and derived lead amplitudes ranged from 0.78 to 0.99 for limb leads, and using the Dower values ranged from 0.81 to 0.94 for the precordial leads. These results show a clear association between the measured QT interval dispersion and the variation in ECG lead amplitudes derived from a simple heart vector model of repolarization with no regional information. Therefore, measured QT dispersion is related mostly to a projection effect and is not a true measure of repolarization dispersion. Our existing interpretation of QT dispersion must be reexamined, and other measurements that provide true repolarization dispersion data investigated. PMID- 11025897 TI - A cosmetic approach for pectoral pacemaker implantation in young girls. AB - Pectoral placement of pacemaker generators, combined with use of a redundant intravascular lead portion, reduces the need for endocardial lead advancement during growth in children. While the use of small generators and submuscular pockets has contributed to cosmetic acceptability, the conventional subclavicular incision may occasionally form a keloid scar that is unacceptable in young girls. A modified implantation technique was used in five girls (age 2.6-13.3 years) during implantation of VDD (n = 2), VVIR (n = 2), and DDDR (n = 1) pacemakers. A 5-cm incision was made in the axilla along the line of the pectoralis major and dissection was continued below the muscle to create a pocket for the generator. Subclavian vein puncture was performed from the axillary incision and beneath the pectoralis major muscle using standard or extra long needles with a needle guard. Peel away sheaths were used for lead positioning. The generator was placed in the submuscular pocket and the wound closed with absorbable sutures. At follow-up, pacemaker function was excellent and neither the scars nor pacemakers were visible from the front. In conclusion, the axillary incision with direct subclavian vein puncture from below the pectoralis major muscle offers the advantages of pectoral pacemaker implantation through a single cosmetic incision. PMID- 11025898 TI - Updated appraisal of pacing lead performance from the Danish Pacemaker Register: the reliability of bipolar pacing leads has improved. AB - The Danish Pacemaker Register was established in January 1982, and the 12 implanting centers in Denmark report to the Register on a continuous basis by use of the European Pacemaker Patient Identification Card. As of August 1999, the Register contained data on 33,164 bradycardia, endocardial, and epicardial (n = 159) lead implants performed in Denmark on 27,738 generators in 24,180 patients for a total of 17,988 (54.2%) ventricular unipolar, 5,610 (16.9%) ventricular bipolar, 2,056 (6.2%) atrial unipolar, and 7,242 (21.8%) atrial bipolar leads. Lead failure was defined as need for replacement or surgical abandonment of the lead due to pacing or sensing problems with the exception of lead displacement. The 10-year survival rate for unipolar leads was 97.2 +/- 0.4% (+/- 2 SE) as compared to 79.6 +/- 3.2% for bipolar leads (P < 0.001). The Medtronic 4012, the Telectronics 284, and the Siemens (Pacesetter) 1010T, 105T, and 1050T bipolar lead models were poor performing leads that needed careful monitoring and appropriate replacement. Excluding those five poor performing bipolar leads, the models yielded a 10-year survival rate of 94.3 +/- 2.4% for bipolar leads. The 5 year survival for all bipolar leads implanted after January 18, 1993 (this was the date that the last of the five poor performing bipolar lead models were implanted in Denmark) was 98.7 +/- 0.4% compared with 98.9 +/- 0.3% for all unipolar leads implanted after January 18, 1993 and 86.8 +/- 1.6% for all bipolar leads implanted on or before January 18, 1993. These results indicate a significant improvement in bipolar lead reliability. PMID- 11025899 TI - Can sudden cardiac death be predicted from the T wave of the ECG? A critical examination of T wave alternans and QT interval dispersion. PMID- 11025900 TI - The first P synchronous cardiac pacemaker: restoration of AV synchrony in complete heart block. PMID- 11025901 TI - Why did mode switching occur? PMID- 11025902 TI - Cross-spectral analysis of heart rate and blood pressure modulations. AB - The cross-spectral analysis of heart rate (HR) and blood pressure (BP) variabilities provides "amplitude" and "phase" related measures. Compared to the amplitude measure, that is the baroreflex gain, the phase related measure characterizing the time lag between HR and BP oscillations has been studied to a much lesser extent. A population of 103 patients (73 men, 30 women, aged 53 +/- 12, range 20-82 years) referred for the management of coronary artery disease and/or hypertension were studied. In each subject, electrocardiogram and BP recordings were obtained in the supine and sitting positions of 5 minutes of rest (spontaneous respiration), 3 minutes of controlled respiration at 0.1 Hz (slow controlled respiration), and 3 minutes of controlled respiration at 0.33 Hz (fast controlled respiration). The frequency of maximum coherence (above the arbitrary threshold of 0.5) of BP and RR interval variabilities was searched between 0.033 0.133 Hz and 0.200-0.400 Hz to obtain baroreflex gain and phase shift in low and high frequency bands, respectively. Mean phase shifts of -79.1 and -67.0 degrees (-2.4 and -2.1 s) were found during slow-controlled respiration in the supine and sitting body positions, respectively. The mean phase shift between systolic BP and RR interval in the low frequency band was found between 83 and -109 degrees for body positions and respiration regimes. The actual baroreflex related time lag between systolic BP and RR variations was found between 3.5 and 5.1 seconds. The study concludes that the appropriate, and not always easy, selection of the frequency of maximum coherence between BP and HR oscillation is crucial for an accurate cross-spectral assessment of baroreflex sensitivity. PMID- 11025903 TI - Potentially fatal interaction between azithromycin and disopyramide. AB - A patient on disopyramide developed disopyramide toxicity when treated concurrently with azithromycin. Evidence of toxicity included an elevated serum disopyramide level and ventricular tachycardia requiring cardioversion. The azalide antibiotic presumably inhibited dealkylation of disopyramide to its major metabolite, mono-N-dealkyldisopyramide. Physicians should avoid using azithromycin in patients on disopyramide. If this drug combination is unavoidable, disopyramide levels must be closely monitored. PMID- 11025904 TI - Ventricular pacing failure after a single oral dose of pilsicainide in a patient with a permanent pacemaker and paroxysmal atrial fibrillation. AB - A single oral dose of pilsicainide, a Class Ic antiarrhythmic drug, is a widely used and highly effective therapy for termination of recent onset atrial fibrillation. We report on a patient in which ventricular pacing failure occurred immediately after a single oral dose of pilsicainide. It did not exhibit a parallel relationship between the change in the pacing threshold and plasma concentration of pilsicainide, and the recovery period for the ventricular pacing threshold was longer than that of the plasma concentration of pilsicainide in this patient. Careful attention should be paid when a single oral dose of pilsicainide for termination of recent onset atrial fibrillation is used in patients with permanent pacemakers. PMID- 11025905 TI - Implantation of a cardiac resynchronization system for idiopathic dilated cardiomyopathy in a patient with persistent left superior vena cava using an experimental lead for left ventricular stimulation. AB - A persistent left superior vena cava (PLSVC) was discovered at the implantation of a cardiac resynchronization system in a woman with an idiopathic dilated cardiomyopathy. Standard leads were used to obtain right ventricular and right atrial (RA) stimulation according to a formerly described technique. The left ventricle was stimulated through the posterolateral vein of the heart by a novel lead design to be used over a guidewire for placement. Despite expected difficulties in this anatomic situation, the research lead was positioned in 23 minutes. PMID- 11025906 TI - Successful radiofrequency catheter ablation of left ventricular tachycardia following surgical correction of tetralogy of Fallot. AB - A 13-year-old boy had recurrent ventricular tachycardia following surgical correction of tetralogy of Fallot. Pharmacological management with multiple antiarrhythmic drugs failed, and the patient underwent electrophysiological evaluation. A reentrant tachycardia involving a slowly conducting region in the left ventricular outflow tract was demonstrated. We performed radiofrequency catheter ablation after identifying the essential components of the reentrant circuit. The ventricular tachycardia could no longer be induced and did not recur spontaneously during a 13-month follow-up. PMID- 11025907 TI - Inappropriate pacing due to autoperpetuation of the ventricular rate stabilization algorithm: a manifestation of T wave oversensing by ICDs. AB - Preventive pacing algorithms designed to eliminate the pause that follows a premature ventricular depolarization have been incorporated in current implantable defibrillators. We report a patient in whom intermittent T wave oversensing frequently invoked the ventricular rate stabilization algorithm and resulted in periods of inappropriate VVI pacing. The problem was solved by decreasing the maximum sensitivity from 0.3 mV to 0.45 mV. Implant testing had revealed adequate sensing of ventricular fibrillation with a maximum sensitivity of 1.2 mV. PMID- 11025908 TI - DDD pacing and interatrial conduction block: importance of optimal AV interval setting. AB - The case of a 83-year-old patient undergoing DDD pacemaker implantation for sick sinus syndrome with postimplant detection of advanced interatrial conduction block is described. At nominal AV interval programming values (175 ms), absence of P wave following an atrial spike was observed, and the presence of an interatrial conduction disturbance was demonstrated by a Doppler transmitral flow pattern analysis and transesophageal ECG recording. AV interval lengthening up to 300 ms resulted in proper timing of atrial and ventricular contractions. Awaiting for conclusive data about biatrial pacing, interatrial conduction blocks can be managed in some cases by proper programming of conventional DDD systems. PMID- 11025909 TI - Stability and course of personality disorders: the need to consider comorbidities and continuities between axis I psychiatric disorders and axis II personality disorders. AB - The literature pertaining to the stability and course of personality disorders is briefly reviewed. Available data suggest that PD diagnoses demonstrate only moderate stability and that--although generally associated with a plethora of negative outcomes--they can show improvement over time. This paper highlights the pervasiveness of diagnostic co-occurrence and its implications for continued investigation of the question of PD stability. In addition to examining the stability (and outcome) of PDs, longitudinal studies need to consider continuities between diagnoses. The Collaborative Longitudinal Personality Disorder Study (CLPS) is described briefly vis-a-vis some of these major issues. PMID- 11025910 TI - Functional impairment and cognition in bipolar disorder. AB - Bipolar disorder is a common, chronic and severe mental disorder, affecting approximately 2% of the adult population. Bipolar disorder causes substantial psychosocial morbidity that frequently affects the patient's marriage, children, occupation, and other aspects of the patient's life. Few studies have examined the functional impairment in patients with affective illness. Earlier outcome studies of mania reported favorable long-term outcomes. However, modern outcome studies have found that a majority of bipolar patients evidence high rates of functional impairment. These low reports of functional recovery rates are particularly surprising. The basis for such limited functional recovery is not entirely clear. Factors associated with functional dysfunction include presence of inter-episode symptoms, neuroleptic treatment, lower social economic class, and lower premorbid function. Cognitive dysfunction, a symptom domain of schizophrenia, has been identified as an important measure of outcome in the treatment of schizophrenia. Recently, there has been some suggestion that there may be impaired neuropsychological performance in euthymic patients with recurring mood disorders. Whether impaired neuropsychological performance in associated with the functional impairment in bipolar patients who have achieved syndromal recovery is an intriguing question. The literature on functional impairment and cognition in bipolar disorder is reviewed. PMID- 11025911 TI - Successfully treating aggression in mentally ill prison inmates. AB - The population of prison inmates in the United States is rapidly growing; in 1994 it was estimated that 1.7 million Americans were incarcerated. Changes in the complexion of the prison population highlight the need for successful mental health interventions for special populations. Specifically, increasing numbers of women, middle aged inmates, physically ill inmates, and inmates with severe mental illness have challenged an already taxed system. Many inmates have complex pathology in which substance abuse, psychosis, affective disorders, personality disorders and medical illnesses play overlapping roles. One of the most challenging aspects of care involves management of inmates who are aggressive to themselves or others. This article discusses the approach to managing inmates with aggressive behavior and complex mental health issues. Specific diagnoses are discussed as are the general approaches best used to assess new onset violent behavior in a mentally ill prison inmate. PMID- 11025912 TI - Suicide and aging: international perspectives. AB - From Durkheim to Kraepelin, suicide has been recognized as a social and psychiatric ill. Among clinical scientists in the United States, interest in suicide grew substantially in the 1980's as rates among older adults increased. However, major advances in the science of mental health seem unlikely to reduce the prevalence of suicide as long as case recognition at the community level continues to be problematic. Public policy promoted to reduce social risk factors coupled with greater attention to psychopathology is the logical outgrowth from the most recent data. Differences in suicide rates by nation add weight to the argument. PMID- 11025913 TI - Defending the unabomber: anosognosia in schizophrenia. AB - The use of recent psychiatric research in the defense of the 'Unabomber' (United States vs. Theodore Kaczynski) is a compelling example of how the gap between research and practice can have profound consequences on the practice of forensic psychiatry, psychology and the judicial process. In this case, educating the lawyers and the court about the research on poor insight in schizophrenia changed the defense strategy and ultimately the course of the trial. PMID- 11025914 TI - Treatment refractory schizophrenia. AB - Treatment resistance constitutes a significant dilemma in schizophrenia since it affects a substantial number of patients, their families and the health care professionals involved in their care. Nonresponsiveness needs to be approached as a multidimensional syndrome by specifying which symptoms in the spectrum of positive symptoms, negative symptoms, excitement/hostility, cognitive symptoms, and anxiety/depression are failing to respond to treatment. This review presents some of the clinical, demographic and biological correlates of nonresponse, in addition to compliance issues, psychosocial factors or side effects and as-yet untreated comorbidities as a source for nonresponse. The effects of the atypicals clozapine, olanzapine, risperidone and quetiapine as compared to typicals are reviewed using available double-blind studies in this treatment refractory group of schizophrenia patients. The limited number of reports on the comparison of atypical compounds amongst each other are critically presented. Given that a subset of patients still do not respond to these agents, clinicians are using various augmentation strategies. We review studies with augmentation strategies which remain difficult to interpret given the open label and uncontrolled nature of most of these studies. PMID- 11025915 TI - [Internet--an introduction]. AB - The internet is an excellent medium for storing and quickly disseminating large amounts of information on a global basis. Modern medicine requires a rapid access to information including clinical data from medical records, bibliographic databases, knowledge bases and nomenclature databases. This is especially true for university hospitals and medical schools for training as well as for fundamental and clinical research for diagnosis and therapeutic purposes. Therefore, the internet with its constantly increasing offers plays a more and more important role in scientific research. This review focuses on how developments of this communication technology have become a useful educational resource in medicine, and describes modest ideas in computer network use. Internet basic resources are electronic mailing (e-mail), discussion groups, file transfer, and browsing the World Wide Web (WWW). PMID- 11025916 TI - [Vasculitis 1998: new aspects in diagnosis and therapy]. AB - The Chapel Hill conference established the most actual classification of systemic vasculitides. Disease activity and organ damage were standardized by means of a new scoring system (BVAS; VDI). Bacterial infection is frequent during the active phase of vasculitis. The measurement of serum procalcitonin helps to differentiate between disease activity and infection. In a recent study the European vasculitis group (EC-BCR project) found that the presence of C-ANCA/PR 3 ANCA or P-ANCA/MPO-ANCA is as reliable as histological proof of vasculitis. In rare cases ANCA is positive in diseases other than vasculitis, which has to be kept in mind for diagnosis and therapy. The association between ANCA and IgA nephropathy is exciting. The occurrence of IgA-GN in patients with Wegener's granulomatosis is usually a secondary event whereas in patients with rapidly progressive IgA-GN microscopic polyangiitis follows. Many observations support the view that ANCA-positive vasculitides are T-cell mediated diseases. Our investigations demonstrate that neutrophils express class II antigens during active disease and are able to trigger T-cell activation via infectious stimuli. With regard to therapy of ANCA-positive vasculitides several strategies are currently being examined in different European countries (EUVAS) with the aim to limit cyclophosphamide to the active stage of the disease. PMID- 11025917 TI - [Health consequences of passive smoking]. AB - Environmental tobacco smoking (ETS) represents a main risk factor for the generation of diseases of the respiratory tract and of the cardiovascular system in spite of opposite assertions. ETS enhances the risk of lung cancer by a factor of 2-3. New born and small children (< 2 years of life) are at high risk if they live within this period of time in a household together with a maternal more than fraternal smoking. Endothelial cells of the blood vessels were already damaged during the first month of life of passive smoking children, and these defects can be measured during the first decade of life. ETS over a period of more than ten years changes the intima/media ratio by enhancing the thickness of the vessel wall. In addition, poor health behaviour is seen in households of smokers because the behaviour of the parents is transferred to that of their children, and this behaviour is the starting point of further health risks and damages. The represented data should firstly lead to enhancing primary smoking prevention among children and young people, and secondly to organizing programs against ETS at the workplace and in public buildings and at last also in the private home. More than up to date, the non-smoker must be informed about the risks and damages of ETS. PMID- 11025919 TI - Intramuscular hemangioma mimicking a bone tumor on plain film. AB - We report a case of intramuscular hemangioma of the leg in a young man showing a bone tumor-like appearance on plain film. Correct preoperative diagnosis was obtained by MR imaging. PMID- 11025918 TI - [Selected secondary reconstructive procedures for improvement of urinary incontinence in bladder exstrophy and neurogenic bladder dysfunction in childhood]. AB - Partial or complete urinary and stool incontinence due to malformation of the genito-urinary tract and the pelvic floor despite of several operative reconstructions is the most important handicap in the patients life. Often this problems seems to be unsoluble. In recent time we secondarily reconstructed 7 patients suffering from urinary incontinence: 1 girl and 4 boys with bladder ekstrophy; 1 boy with a complex anomaly with menigomyelocele, sinus urogenitalis, single kidney with vesicorenal reflux and neurogene bladder; 1 boy with complex anomaly of the pelvis and the lower limbs with duplication of the bladder with an ekstrophic left part. All patients underwent multiple operative trials of reconstruction. Until that time all patients suffered from complete urinary incontinence. At the age of 5 to 14 years we performed the secondary reconstruction: bladder-neck-plasty and ileumaugmentation (3 patients), closure of the bladder-neck, ileumaugmentation and a continent appendicostoma (Mitrofanoff's method)), Mainz-I-pouch and a continent appendicostoma respectively ileostoma (Monti's technique) (2 patients), Mainz-II-pouch. With a normal bladder-capacity all patients are completely continent postoperatively; one patient has regained partial continence. The emptying of the bladder is carried out by clean intermittent catheterism (CIC) with the exception of the one patient with the Mainz-II-pouch. Even in patients with complex anomalies of the pelvic floor and the genitourinary tract complete urinary continence is possible in consequence of recently developed operative techniques. Because of a high rate of complications we reject the primary use of artificial sphincter systems for children. PMID- 11025920 TI - [Effective dose and dose to the crystalline lens during angiographic procedures]. AB - Interventional radiology presents the highest dose levels for the staff and for the patient. For a total of 34 angiographic examinations the doses at the forehead and at the neck of a radiologist have been measured. The effective dose and the dose to the eye lens have been estimated over a period of one year. Doses to the lens of the eye close to the limit of 150 mSv proposed by the ICRP have been observed. There is an imminent need for the radiologist to wear also lead glasses to avoid formation of cataracts at long term. PMID- 11025921 TI - Sedation, analgesia and anesthesia for interventional radiological procedures in adults. Part I. Survey of interventional radiological practice in Belgium. AB - A questionnaire was mailed to 217 interventional radiologists to evaluate current practice in analgesia and sedation in adults. Response rate was 15% (33/217). Diagnostic angiography was performed with local anesthesia in 94% to 99%; for PTA, local thrombolysis or stent placement, light sedation was added in 0.1%. Premedication was given in 43% of diagnostic angiographies and in 68% of therapeutic procedures. Radiologists consulted an anesthesiologist before administration of intravenous sedation, always in 54% of cases, occasionally in 19% and never in 27%. General anesthesia with artificial ventilation was applied in 56% of TIPS, in 70% of aortic stent grafting and in 82% of neuroradiological interventions. Intravenous sedation was applied given in 53% of percutaneous biliary drainage, in 42% of bile duct dilatation or stenting, in 40% of percutaneous nephrostomy and in 72% of ureteral balloon dilatation. Patient monitoring during an interventional procedure was always carried out by an anesthesiologist in 52% of cases. 21% of radiologists never visited the patient before a therapeutic procedure, and 36% never did so after completion of a procedure. This survey showed that high standard practice of sedation and analgesia, with the assistance of anesthesiologists, is underused by interventional radiologists in Belgium. PMID- 11025923 TI - Doppler sonography in breast pathology. AB - Breast sonography has been a standard complementary procedure to mammography for more than 10 years. In recent years, a tremendous technological evolution has taken place both at the level of the equipment, electronics and software, and in probe technology. This has resulted in a substantial increase in image quality (contrast ans spatial resolution) and in Doppler sensitivity. The use of linear probes of at least 7 MHz is mandatory, and frequencies of over 10 MHz are becoming standard now. Sonography of the breast not only permits differentiation between solid and cystic lesion, but also gives additional morphological details which enable differentiation between benign and malignant lesions. US increases both sensitivity and specificity of the breast exam when combined with X-ray mammography. It is also the method of choice for guiding interventional procedures, such as hook wire placement, FNAC, or core biopsy. Doppler has been used since the seventies (continuous wave Doppler), but more recently the use of color Doppler has proven to be valuable in the study of the vascularity of breast lesions. PMID- 11025922 TI - Sedation, analgesia and anesthesia for interventional radiological procedures in adults. Part II. Recommendations for interventional radiologists. AB - Benzodiazepines are given orally as a premedication before an interventional radiological procedure. Local analgesia is achieved by drugs such as lidocaine, bupivacaine or ropivacaine. General analgesia is obtained by non opioid analgesics and opioid narcotics. For intravenous sedation, benzodiazepines such as ketamine or propofol should be administered under the supervision of an anesthesiologist. A preprocedure consultation with the anesthesiologist is recommended. Monitoring equipments, drugs and nursing staff assistance should be provided in the interventional suite. Vital signs should be monitored for several hours until patient's discharge. Close collaboration between anesthesiologists and interventional radiologists is a prerequisite for achieving high standard sedation and analgesia. PMID- 11025924 TI - Review of MR mammography techniques. AB - Many of the new techniques of MRI have promising characteristics for the further improvement of sensitivity and specificity but there remains a large place for clinical trials to evaluate the practical value of the different new approaches. The authors review the general MRI rules that determine image quality and comment on the use-fullness of innovations in MR mammography. PMID- 11025925 TI - Indications for MR mammography. A Belgian study. AB - The problem of dynamic MR mammography remains its low specificity. Therefore, the examination is only useful when it is performed as a complementary examination and only for good indications. The aim of the study was to analyse the indications for MR mammography in 12 Belgian hospitals and to evaluate whether MRI was useful for these indications. The two most frequent reasons for which MR mammography was performed in these hospitals were a diagnostic problem after clinical examination, mammography and ultrasound, and differentiation between post-therapeutic changes and recurrent disease, both of which are good indications. MRI examinations in the 12 hospitals were useful in patients receiving the examination for a good indication. However, in the group of the known poor indications we see much false positive MRI results. PMID- 11025926 TI - Images in clinical radiology. Bicipitoradial bursitis. PMID- 11025927 TI - Beyond white and middle class: cultural variations in families--assessments, processes, and policies. AB - The goal of this special issue on cultural variations in families--assessments, processes, and policies--is to increase our understanding of families of different ethnic, racial, and class backgrounds. Articles in this issue represent a range of cultural groups, illustrate a pattern of both similarities and differences across groups, and reflect the recent emphasis on processes that account for variation in families of different ethnic backgrounds. The articles focus on strengths of families of different ethnic backgrounds in contrast to earlier deficit models. Finally, theoretical models that recognize within-group variability of different ethnic, racial, and cultural groups are illustrated in these articles. PMID- 11025928 TI - Immigrant-Chinese and Euro-American parents' physical closeness with young children: themes of family relatedness. AB - This study examined cultural differences in the expression and meaning of physical closeness. Findings indicated that immigrant-Chinese parents, as compared with Euro-American parents, sleep in closer proximity with their children; more often view independence as children growing with the family; are more likely to emphasize the family unit; and place greater importance on inhibition of expression, adherence to correct values, and hierarchy of relations. Euro-Americans, by contrast, are more accepting of nudity; place more emphasis on psychological benefits of physical closeness and on the child's expression of wants and feelings; more often view independence as celebrating the child's distinctiveness; and place greater importance on intimacy, pleasure, and spousal exclusiveness. The authors suggest that closeness is characterized by an overarching theme of harmony in immigrant-Chinese families and by an overarching theme of romance in Euro-American families. PMID- 11025929 TI - Moderating effect of perceived amount of family conflict on the relation between home environmental processes and the well-being of adolescents. AB - This study examined the extent that perceived amount of conflict in the family moderates the relation between proximal aspects of the home environment and adolescent well-being. Regression models consisting of Home Observation for Measurement of the Environment (HOME) subscale scores (Responsivity, Learning Materials, Variety of Experiences), family conflict, and the interaction between HOME scores and family conflict were tested for each of 6 dependent measures: academic achievement, task orientation, being considerate, self-efficacy as it pertains to school, self-efficacy as it pertains to family, and school grades. Results indicated moderation in all 3 ethnic groups examined (European Americans, African Americans, and Chinese Americans). For all 3 groups, relations were stronger in families with high conflict, suggesting a heightened sensitivity to social exchanges and events within the family. PMID- 11025930 TI - A mediational model of the impact of interparental conflict on child adjustment in a multiethnic, low-income sample. AB - Path analysis was used to determine whether the effects of interparental conflict on children's depression and conduct disorder are mediated by 3 dimensions of parenting: acceptance, inconsistent discipline, and hostile control. The study extends the literature by testing this mediational model with a low-income, predominantly ethnic minority sample of preadolescent children and by examining the effects of multiple dimensions of interparental conflict from the child's perspective. Results supported the mediational model when analyses were based on child's reports of all variables but not when mother's reports were used to assess child depression and conduct problems. Exploratory analyses revealed unique mediational paths associated with conflict frequency and resolution, which were examined along with intensity as distinct dimensions of interparental conflict. PMID- 11025931 TI - Discipline responses: influences of parents' socioeconomic status, ethnicity, beliefs about parenting, stress, and cognitive-emotional processes. AB - Direct and indirect precursors to parents' harsh discipline responses to hypothetical vignettes about child misbehavior were studied with data from 978 parents (59% mothers; 82% European American and 16% African American) of 585 kindergarten-aged children. SEM analyses showed that parents' beliefs about spanking and child aggression and family stress mediated a negative relation between socioeconomic status and discipline. In turn, perception of the child and cognitive-emotional processes (hostile attributions, emotional upset, worry about child's future, available alternative disciplinary strategies, and available preventive strategies) mediated the effect of stress on discipline. Similar relations between ethnicity and discipline were found (African Americans reported harsher discipline), especially among low-income parents. Societally based experiences may lead some parents to rely on accessible and coherent goals in their discipline, whereas others are more reactive. PMID- 11025932 TI - Parental warmth, control, and indulgence and their relations to adjustment in Chinese children: a longitudinal study. AB - A sample of children, initially 12 years old, in the People's Republic of China participated in this 2-year longitudinal study. Data on parental warmth, control, and indulgence were collected from children's self-reports. Information concerning social, academic, and psychological adjustment was obtained from multiple sources. The results indicated that parenting styles might be a function of child gender and change with age. Regression analyses revealed that parenting styles of fathers and mothers predicted different outcomes. Whereas maternal warmth had significant contributions to the prediction of emotional adjustment, paternal warmth significantly predicted later social and school achievement. It was also found that paternal, but not maternal, indulgence significantly predicted children's adjustment difficulties. The contributions of the parenting variables might be moderated by the child's initial conditions. PMID- 11025933 TI - Parenting practices and adolescent depressive symptoms in Chinese American families. AB - This study examined parenting practices and adolescent depressive symptoms among Chinese Americans. First, confirmatory factor analyses revealed that self reported parenting practices by mothers and fathers and adolescent perception of parenting practices loaded adequately on three subscales: Inductive Reasoning, Monitoring, and Harsh Discipline. Second, parents' depressive symptoms were related to disrupted parenting practices, which, in turn, were significantly related to the negative evaluation of these behaviors by the adolescents. Adolescents' perceptions of such parenting practices were significantly associated with their depressive symptoms. Third, the relationships were robust even after parental income, education, and generation status were statistically controlled. Overall, the relationships between parenting practices and adolescent depressive symptoms among Chinese Americans seemed to echo those found among European Americans. PMID- 11025935 TI - Adolescent-parent interactions in middle-class African American families: longitudinal change and contextual variations. AB - Family interactions were examined longitudinally over 2 years in 79 middle-class African American families with early adolescents. Mothers and adolescents (as well as fathers and adolescents and triads in 2-parent families) were videotaped discussing a conflict for 10 min. A macro-coding system (J. G. Smetana, J. Yau, A. Restrepo, & J. L. Braeges, 1991) was modified to be culturally sensitive to African American families. Interaction ratings were reduced, through principal components analyses, into composite variables. After control for family income, mothers' communication in triadic interactions became less positive over time. Both mothers' and fathers' communication was more positive in dyadic than triadic interactions. In triadic interactions, mothers validated sons more than daughters, and in dyadic interactions with either parent, boys were more receptive to parents than were girls. Findings extend previous research on adolescent-parent relationships to African American families. PMID- 11025934 TI - Patterns of family functioning and adolescent outcomes among urban African American and Mexican American families. AB - The relations of patterns of family functioning, prosocial behaviors, and internalizing and externalizing symptoms over time were evaluated among a sample of economically disadvantaged inner-city African American and Mexican American male adolescents. Ethnic group differences for configurations of family functioning over time, levels of prosocial and problem behaviors, and relations of family functioning to risk were found. Among both ethnic groups, exceptionally functioning families provided a protective effect against risk. Overall, African American youth had stronger attitudes toward school and higher educational aspirations than Mexican American youth. Unlike previous investigations, once socioeconomic status was controlled, no differences were found for either internalizing or externalizing problems between the 2 groups. The importance of considering socioeconomic status and community context when evaluating minority parenting and family functioning is discussed. PMID- 11025936 TI - Effects of parental marital status, income, and family functioning on African American adolescent self-esteem. AB - This study examined the effects of marital status, family income, and family functioning on African American adolescents' self-esteem. One hundred sixteen adolescents participated, 64% of whom were female. Compared with boys with nonmarried parents, boys with married parents had higher overall self-esteem, even when family income and family functioning were controlled. Parental marital status had no effect on girls' self-esteem. Family functioning was a very strong predictor of self-esteem for both sexes. However, family relational factors were more important to girls' self-esteem, whereas structural and growth factors were more important for boys. It was concluded that African American adolescent boys with nonmarried parents are at risk for developing low self-esteem compared with other African American adolescents, but a more controlled and structured environment may buffer the effects of having nonmarried parents. PMID- 11025937 TI - Differential parental treatment predicts achievement and self-perceptions in two cultural contexts. AB - This study examined 148 Asian American and European American late adolescents' perceptions of differential affection and control by mothers and fathers as predictors of academic achievement and self-perceptions of intellectual ability and global self-worth. Overall, analyses generally confirmed the hypotheses that the more differentially favorable the treatment (more affection or less control) or the less differential treatment (above and beyond which sibling was favored) reported in the home, the more positive late adolescents' outcomes. Perceptions of differential parental treatment predicted up to 13% of the variance in achievement and self-perceptions. Several findings were moderated by ethnicity or gender. Finally, and perhaps most importantly, perceptions of differential parental treatment predicted a significant and unique amount of variance in outcomes beyond that predicted by perceptions of absolute levels of affection and control. PMID- 11025938 TI - Black and white girls' racial preferences in media and peer choices and the role of socialization for black girls. AB - This exploratory research compared Black and White girls' racial preferences as exhibited through their media (music and television) and peer choices. The sample included 140 8- and 9-year-old Black and White girls of various socioeconomic levels. Findings suggested that both Black and White girls have more Black music preferences than White or no-race music preferences. Also, both Black and White girls made more White television program choices than Black or no-race choices. In their peer selections, all girls preferred same-race peers. Black mothers who engaged in racial socialization practices had girls who were more likely to prefer Black music and television to the other categories. Further, Black mothers who promoted more cultural distance and mothers who were poor had girls with more same-race peer preferences. PMID- 11025939 TI - On the formation of the temporomandibular joint cavity in the human fetus. AB - The purpose of this study was to clarify the mechanism of articular cavity formation by means of observing the temporomandibular joint (TMJ) in human fetuses (9th-30th gestational weeks) using light microscope. In the 9th-11th week, although neither cavity had been formed, several small blood vessels running postero-anteriorly on the lower surface of the articular disk that would be the future lower cavity were recognized. However, such blood vessels were not seen in the future upper cavity beneath the glenoid fossa. In the 12th week, when the lower cavity formation commenced, the cavity was filled with blood corpuscles, and a series of apertures was observed throughout the condyle, from the posterior portion to a part of the anterior portion. Tracing posteriorly, the lower cavity diminished gradually until it had at last united to one blood vessel. On the other hand, in upper cavity formation, which was recognized only at the posterior portion of the TMJ, no blood corpuscles were seen in the upper cavity but several small clusters of collagen fibers. Blood corpuscles in the lower cavity disappeared after the formation of synovial membrane (20th week) and vascular canals in the condyle (21st week). From these findings, the mechanism of lower cavity formation appears to differ from that of upper cavity formation. Lower cavity formation involved small blood vessels running postero-anteriorly on the lower surface of the articular disk. As for upper cavitation, there was no evidence of blood vessels, which would suggest that upper cavitation depends on another mechanism of formation. PMID- 11025940 TI - Histochemical studies on glycosaminoglycans in Bruch's membrane of postnatal rat eyes. AB - Localization of glycosaminoglycans (GAG) in Bruch's membrane of postnatal rat eyeballs was examined histochemically. Fixed eyeballs from postnatal rats (ages 5 days and 8 weeks) were routinely processed and embedded in paraffin wax or Quetol 651 resin. Paraffin-embedded tissue sections were stained with hematoxylin and eosin or sensitized high iron diamine procedure in combination with selective methods such as GAG-degrading enzyme digestions and/or a chemical modification, and examined by light microscopy. Quetol 651-embedded ultrathin sections were stained with heavy metals and examined by electron microscopy. In rats at postnatal day 5, Bruch's membrane contained mainly chondroitin sulfate (CS) and heparan sulfate (HS). In contrast, at 8 weeks after birth the membrane included a large amount of dermatan sulfate (DS) and HS. According to electron microscopic findings, Bruch's membrane on day 5 consisted of only 3 layers without a central elastic layer. However, at 8 weeks after birth the membrane was constructed of 5 layers. These findings suggested that the difference in GAG molecular species in the membranes at 5 days and at 8 weeks after birth could be correlated with the development and maturation of the collagenous layer in Bruch's membrane. Moreover, maturation of Bruch's membrane may contributes to the architectural stabilization of the outer portions of the photoreceptor cells. PMID- 11025941 TI - On a complex anastomosis of the glossopharyngeal nerve in humans. AB - The glossopharyngeal nerve shows anastomoses with the facial nerve and the sympathetic nervous system. One anastomosis extends from the interconnected stylopharyngeal branches, immediately after having perforated the muscle towards the base of skull. Cranially, varying targets of the ascending nerve can be discriminated: 1) The temporal bone. 2) The facial nerve. 3) The sympathetic nerve plexus of the internal carotid nerve. This complex anastomosis was now studied under the dissecting microscope in more detail. The investigation revealed a more complicated distribution pattern of the anastomotic nerve than previously assumed, i.e. the existence of a solitary ascending branch could only be proved in a minority of cases (seven of twenty individuals). In the majority, a delicate nerve plexus could be visualized (thirteen of twenty individuals). In the cases of an anastomosis with the facial nerve, the stylohyoid branch was observed to be the main target of the ascending nerve. Also, connections with the internal carotid nerve were seen. In addition, delicate endings of the branches were demonstrated ramifying in the styloid process or penetrating the temporal bone at other sites. The histological demonstration of ganglion cells within the ascending nerve or nerve plexus suggests parasympathetic and sensoric functions for this anastomosis. PMID- 11025942 TI - [Experimental analysis of initial and appositional mineralization of biological hard tissues in bisphosphonate-affected animals]. PMID- 11025943 TI - [Morphological changes of the mandibula with loss of the teeth--observation of the trabecular bone using micro-CT]. AB - The jaw bones receive complicated forces not only through the muscles, but through the teeth directly. Therefore, it is thought that the morphology of the jaw bones are greatly effected by the oral conditions. In this paper, the morphological changes of the mandibula with loss of the teeth are descrived. Furthermore, the three dimensional changes of the trabecular bones observed by micro-CT are shown. The jaw bones are of complex composition since they contain the alveolar sockets that encase the dental roots. The structures of the trabecular bone of the mandibula are constructed in order to support the teeth. The trabecular bones appear to have plate-like shape rather than rod-like shape in the three dimensional observation using micro-CT. With loss of the teeth, the height of the mandibula is rapidly shortened. And in the internal structures, both width and volume of the trabecular bone within the substantia spongiosa are reduced and the trabecular bones run in the irregular course. In addition, the plate-like shaped trabecular bones tend to transform into rod-like shaped ones. Based on these findings, it is suggested that maintenance of the intact teeth is important to keep the normal morphology of the mandibula. PMID- 11025944 TI - [Cementum formation in rat molar roots]. AB - Cementum is the calcified tissue covering roots of teeth and serves as attachment sites of the periodontal ligament. Although recent studies have suggested that extracellular matrix of cementum is very similar to that of bone, cementogenesis on a biological basis is still poorly understood. There are variations in the distribution and mineral contents of cementum depending on animal age, tooth species and position within the tooth roots. This paper reviews the formation and age-related changes of cellular and acellular cementum in rat molar roots, and discusses the effect of mechanical stress to the cementum formation. PMID- 11025945 TI - [Role of PTHrP in mouse molar development]. PMID- 11025946 TI - [Morphological basis of periodontal nerve endings]. AB - The periodontal ligament, a dense collagenous tissue between tooth and alveolar bone, receives a rich sensory nerve supply, and contains two kinds of sensory receptors; nociceptor and mechanoreceptor. The mechanical stimuli for teeth can evoke various oral reflexes, which facilitate mastication via the periodontal mechanoreceptors. In spite of many reports on the periodontal sensory receptors, recent studies have revealed that the Ruffini endings, categorized as low threshold slowly adapting type II (SA II) stretch receptors, are primary mechanoreceptors in the periodontal ligament. This paper summarized recent findings on the morphological features and developmental aspects of the periodontal Ruffini endings. PMID- 11025947 TI - [Ultrastructure of clathrin sheets and cytoskeleton of podosomes on the cytoplasmic side of ventral membranes of cultured osteoclasts]. PMID- 11025948 TI - [Causes of renal dysfunction during the anhepatic phase in patients for orthotopic liver transplantation and their postoperative renal function]. AB - This study was carried out to clarify causes of renal dysfunction during the anhepatic phase in orthotopic liver transplantation (OLT) with venovenous bypass and to show how the deterioration impacted on postoperative renal function. The 44 consecutive patients with normal preoperative renal function who underwent OLT in Royal Prince Alfred Hospital were classified into two groups according to creatinine clearance (Ccr) during the anhepatic phase. Group 1 consisted of 27 patients whose Ccr levels levels were kept above 60 ml.min-1.m-2 and group 2 consisted of 17 patients under 60 ml.min-1.m-2. In group 2, preoperative International Normalized Ratio for prothrombin was higher and blood transfusion volume before revascularization was significantly lager than group 1. There were significant differences in haemodynamics just before revascularization (mean arterial pressure 95 +/- 14 vs 83 +/- 14 mmHg, pulmonary artery wedge pressure 16 +/- 5 vs 11 +/- 5 mmHg, cardiac index 4.6 +/- 1.3 vs 4.0 +/- 0.9 l.min-1.m-2, group 1 vs group 2, mean +/- SD, P < 0.05). Serum creatinine levels in group 2 were significantly higher postoperatively. It is suggested that bleeding due to insufficient preoperative coagulability caused haemodynamic instability which deteriorated renal function during the anhepatic phase and the deterioration impacted on postoperative renal function. PMID- 11025949 TI - [The effect of acetate ringer solution, 6% hydroxyethyl starch saline and 20% mannitol solution on the serum concentration of propofol continuously infused]. AB - Changes in serum concentrations of propofol after administration of three different fluids were investigated in 42 scheduled surgical patients. Anesthesia was induced with propofol 2 mg.kg-1 and maintained with constant rate infusion of propofol 6 mg.kg-1.hr-1. After achieving a stable depth of anesthesia, 5 ml.kg-1 of acetate Ringer's solution, 6% hydroxyethyl starch saline solution or 20% mannitol solution was infused in 15 minutes. Blood samples each 2 ml were taken before and 0, 5, 15, 30 and 60 minutes after fluid treatment. We measured hemoglobin and hematocrit of the samples for calculating the dilution rate of the plasma with infusion treatment, and determined the serum concentration of propofol by HPLC-spectrofluorometry. After administration of each fluid, the serum concentrations of propofol decreased significantly to 17 +/- 15, 25 +/- 10 and 35 +/- 8%, respectively (mean +/- SEM). The dilution rate of the plasma from the fractional change in blood hemoglobin increased to 0.08 +/- 0.02, 0.24 +/- 0.03, and 0.36 +/- 0.03, respectively. Administration of mannitol might markedly increase distribution volume of propofol, and this can be attributed to osmotic action of mannitol and resultant expansion of extracellular fluid volume. The results of the present investigation suggest that this pharmacokinetic change decreased the concentration of propofol more significantly in mannitol treatment patients than in Ringer's solution or 6% hydroxyethyl starch saline treatment patients. PMID- 11025950 TI - [Clinical evaluation of combined spinal-epidural anesthesia by needle-through needle approach in 485 surgical cases]. AB - We analyzed retrospectively the technical and clinical consequences of combined spinal-epidural anesthesia by needle-through-needle approach over the last two years. A Tuohy-type 18-gauge epidural needle (Perican; B. Braun Co.) and long pencil-pointed 27-gauge spinal needle (Whitacre; Becton-Dikinson Co.) were selected. Spinal anesthesia was administered with 0.5% tetracaine. A total of 485 anesthesia cases included 144 cases for lower abdominal, 193 cases for gynecological and 148 cases for orthopedic surgeries. The successful subarachnoid puncture with only one attempt was recorded in 89% of abdominal, 71% of gynecological and 72% of orthopedic cases. On the other hand, in three (0.6%) cases even with several attempts, the puncture was not possible. Inadvertent dural puncture and subarachnoid catheterization occurred in six (1.2%) and four (0.6%) cases, respectively. Inadequate spinal anesthesia was supplemented with epidural anesthesia in 13% of abdominal, 21% of gynecological and 7% of orthopedic cases. No serious complication occurred. We conclude that this needle through-needle approach facilitates subarachnoid puncture with an ultra-fine spinal needle and subsequent epidural catheterization serves for supplemental and post-operative analgesia unless inappropriate subarachnoid indwelling occurs. PMID- 11025951 TI - [Hemodynamic effects of propofol as an anesthesia induction agent in hyperthyroidism patients on chronic beta-blocker]. AB - This study was designed to investigate the effect of propofol on the heart rate and blood pressure in the patients on chronic beta-blocker. Seventy two hyperthyroidism patients scheduled for subtotal thyroidectomy were enrolled. Thirty six patients who were on chronic beta-blocker received either propofol (group beta-P) or thiamylal (group beta-T) as an anesthesia induction agent. In control groups, patients who were not on beta-blocker also received either propofol (group C-P) or thiamylal (group C-T). Anesthesia was maintained with nitrous oxide in oxygen and sevoflurane. Heart rate and systolic blood pressures were monitored and recorded before and during anesthesia. Heart rate decreased significantly in group beta-P compared to three other groups through this study. Significant decreases in systolic blood pressure were observed in the groups beta P and beta-T compared to group C-P. These results suggest that careful attention should be paid to the patients on chronic beta-blocker when propofol was selected as an anesthesia induction agent. PMID- 11025952 TI - [Differences in hemodynamic effects of amrinone, milrinone and olprinon after cardiopulmonary bypass in valvular cardiac surgery]. AB - The differences in hemodynamic effects of amrinone, milrinone and olprinone were evaluated in 46 patients for valvular cardiac surgery after cardiopulmonary bypass (CPB). Patients were randomly allocated to three groups; group A with amrinone infusion (17 patients); group M with milrinone infusion (15 patients); and group O with olprinone infusion (14 patients). Each drug was administrated as a single dose into the venous reservoir of the CPB circuit 15 min prior to the end of emergence from CPB, followed by continuous infusion. Hemodynamic parameters were measured at the time of preCPB (C0), just after the end of CPB (C1), one hour after the termination of CPB (C2) and after the chest closure (C3). Catecholamines were used in order of dopamine, norepinephrine and dobutamine. These doses were modulated to maintain the cardiac index > 3.0 l.min 1.m-2 by each anesthesiologist. Hemodynamic parameters (at C0, C1, C2 and C3) and the doses of cathecholamine (at C1, C2 and C3) were compared among the 3 drugs. The systolic blood pressure in group M was significantly higher than that of group A and group O after chest closure. In group M and A, the systolic blood pressure showed a significant increase after CPB. On the other hand, the systolic blood pressure showed no significant change in group O after CPB. Three drugs showed no significant difference in the dosages of catecholamines used. PMID- 11025953 TI - [Perioperative management of endovascular stent graft placement for abdominal aortic aneurysm]. AB - We retrospectively examined the changes in hemodynamics, oxygen index and renal function along with the complications in 25 patients who had undergone endovascular stent graft placement (ESG) surgery for abdominal aortic aneurysm. During stent graft placement, mean arterial pressure decreased to 58 +/- 8 mmHg by increasing the dose of anesthetics and/or using vasodilators. Except for this intended hypotensive period, mean arterial pressure and heart rate were relatively stable and adequately maintained during surgical manipulation. Oxygenation index was well maintained. A patient with a high preoperative creatinine level underwent prophylactic hemodialysis postoperatively. In other patients except one who died in early postoperative period, both BUN and creatinine levels were kept within normal ranges. Four patients died postoperatively and the causes of the death in two patients are related to the surgical procedure; one with multiple emboli possibly due to released atheloma from the aortic wall during procedure, the other with sepsis due to infected stent graft. Although ESG is a well tolerated procedure, embolism is the most serious complication. Careful preoperative evaluation of the ascending arch and descending aortic wall and monitoring with transcranial doppler are necessary. PMID- 11025954 TI - [Preoperative complications and intraoperative hemodynamic changes in very old patients with femoral neck fractures]. AB - We compared preoperative complications and intraoperative hemodynamic changes in very old patients, 85 years or older, and those with elderly patients aged 70-84 for hip fracture repair. Spinal anesthesia with 0.25 or 0.5% of bupivacaine was performed except for the patients with dementia and/or deformity of the spinal column. The incidence of cardiac disease and anemia was higher in very old patients than in elder patients, and its odds ratios were 2.29 and 3.10, respectively. There is no difference in intraoperative hemodynamic changes between the two groups. Two patients of very old groups had severe intraoperative complications, heart failure and grave arrhythmia, but other patients underwent the operation without severe complication. In conclusion, even in very old patients with hip fracture, spinal anesthesia was performed safely unless patients had serious diseases preoperatively. PMID- 11025955 TI - [Comparison of propofol and isoflurane anesthesia on postoperative nausea, vomiting and pruritus induced by epidural morphine]. AB - We compared propofol-nitrous oxide anesthesia (Group P) with isoflurane-nitrous oxide anesthesia (Group I) on the incidence of postoperative nausea, vomiting and pruritus induced by epidural morphine. Twenty-eight patients for thoracotomy for lung surgeries were randomly assigned either to Group P or Group I. All patients were administrated epidural morphine (4-7 mg.day-1) during and after the operation. The incidence of nausea, vomiting and pruritus was evaluated at the postoperative early (< 9 hour) and late (> 9 hour) periods. In the late postoperative period, in Group P the incidence of nausea and vomiting tended to be low compared with Group I, but the difference was not statistically significant. The incidence of pruritus was not different between the two groups in both early and late periods. PMID- 11025956 TI - [Comparison between total intravenous anesthesia and inhalation anesthesia in the surgery of acute cholecystitis]. AB - We investigated retrospectively the influence of anesthetic methods on the intraoperative managements and postoperative outcomes in 26 patients receiving emergency or early surgery for acute cholecystitis. Fourteen of the 26 patients received total intravenous anesthesia with propofol, fentanyl, and ketamine (PFK group), while the remainder received nitrous oxide and isoflurane or sevoflurane anesthesia (GO group). There were no significant differences between the groups with respect to demographic data. We found no significant differences between the groups in duration of operation and anesthesia, the incidence of intraoperative hypotension, and the use of ephedrine and dopamine during induction and maintenance of anesthesia. After surgery, the PFK group had significantly earlier bowel function than the GO group, with earlier starting of oral intake (54.0 +/- 25.1 vs 89.3 +/- 31.9 hours after surgery; P = 0.026). These data suggest that total intravenous anesthesia by propofol, fentanyl, and ketamine may provide the earlier recovery of bowel function than inhaled anesthesia after emergency or early surgery for acute cholecystitis. PMID- 11025958 TI - [Perioperative management for emergency cesarean section of a patient with reexpansion pulmonary edema]. AB - A 27-year-old woman with 39 week gestation was admitted because of cough and dyspnea accompanied by massive right-sided pleural effusion. Following the right thoracocentesis, about 2000 ml of bloody pleural effusion was drained. Just after the thoracocentesis, the fetal heart rate (FHR) temporarily showed a variable deceleration pattern but the rate was restored spontaneously. One hour later, cough and dyspnea became worse. Changes in FHR pattern indicated the premature separation of the normally implanted placenta. Accordingly, an emergency cesarean section was performed under general anesthesia. Massive foamy tracheal secretion was drained from the tracheal tube during surgery. As her chest X-ray showed signs of pulmonary edema in the right lung, her status was diagnosed as reexpansion pulmonary edema (RPE). She was transferred to the intensive care unit and treated with mechanical ventilation, prednisolone and diuretics. Extubation was performed on the 2nd day after the surgery. On reexpansion of the collapsed lung, it is always necessary to consider not only the hemodynamic changes just after reexpansion but also RPE following reexpansion. PMID- 11025957 TI - [Anesthesia for emergency surgery in 2 extremely low birth weight infants with ileus]. AB - Two extremely-low-birth-weight infants, weighing each 684 and 975 g at birth, underwent emergency surgery because of ileus. Our previous experience with an extremely low birth weight infant, whose hemodynamic control during the surgery had been difficult without administering extra preoperative fluid and transfusion, made us administer enough fluid and transfusion during operation although their urine output was more than 2 ml.kg-1.hr-1. We gave intravenous volume to the present 2 cases before the operations despite the level of preoperative urine output and made their hemodynamic situation more stable during surgeries. We conclude it is very important to administer some amounts of intravenous volume (approximately 8-12 ml.kg-1.hr-1) in extremely low birth weight infants for emergency surgery with ileus regardless of their preoperative urine output. PMID- 11025960 TI - [Severe lightning pain during spinal anesthesia in a patient with diabetic neuropathy]. AB - A 71-year-old woman with diabetic neuropathy who had undergone amputation of the right lower leg for diabetic gangrene 4 years previously, experienced severe lightning pain in both legs during spinal anesthesia. She was scheduled for skin grafting for a burn ulcer on her left foot. Her preoperative physical examination revealed hypesthesia in both legs due to diabetic neuropathy. Spinal anesthesia was performed with a combined spinal-epidural needle at the L 4-5 interspace using 2.0 ml of 0.3% hyperbaric dibucaine in the left lateral position. The region of hypesthesia was spread below Th 4. Ten minutes later, she complained of severe lightning pain in both legs and midazolam 1 mg was administered intravenously against agitation. The severe lightning pain diminished after the administration of pentazocine 7.5 mg intravenously in the recovery room. There was no worsening of neurological findings 5 hours later when the effect of spinal anesthesia disappeared. This clinical picture seems to be different from that of reported cases of phantom limb pain during spinal anesthesia in which severe lightning pain occurred in both legs. This case suggests that patients with diabetic neuropathy might develop severe lightning pain during spinal anesthesia using dibucaine. PMID- 11025959 TI - [Spinal anesthesia for a patient with epidermolysis bullosa hereditaria dystrophica]. AB - A 39-year-old male with epidermolysis bullosa hereditaria dystrophica was scheduled for surgical excision of squamous cell carcinoma in the lower leg. Spinal anesthesia was chosen, because the skin of the puncture site for spinal anesthesia appeared to be dry and scarred, and to have less chance of infection. To avoid shear stress to the skin, we carefully performed anesthetic management including attachment of equipments associated with anesthesia, monitoring and surgery. However, we experienced some troubles and complications associated with anesthetic technique, such as pain on removing a dressing sheet, burning sensations induced by alcoholic stimuli for cold sign test and tent-like traction of the punctured skin on withdrawing the spinal needle. PMID- 11025961 TI - [Right middle cerebral artery occlusion during endovascular embolisation of cerebral aneurysm by Guglielmi detachable coil: a case report]. AB - Right middle cerebral artery embolism by thrombus occurred in 63-year-old female during endovascular embolisation of cerebral aneurysm by Guglielmi detachable coil. Middle cerebral artery occlusion lasted for 60 minute, and haemodynamics was stable during this period. Middle cerebral artery occlusion was detected only by digital subtraction angiography and information from neurosurgeons. Middle cerebral artery blood flow was restored with thrombolytic agents. After tracheal extubation, transient confusion and seizure occurred, but cerebral infarction was not seen in postoperative CT and there was no complication. In case of endovascular embolisation of cerebral aneurysm by Guglielmi detachable coil, systemic heparinization is necessity to decrease the risk of embolism by thrombus, and anesthetist is required to pay attention to cerebral angiography to maintain, close relationship with neurosurgeons and to take measures for cerebral protection. PMID- 11025962 TI - [A case of anaphylaxic shock due to latex glove used on internal examination and on the probe of intrauterine echogram]. AB - A 15 year old female with uterus bicornis bicollis was admitted for operation. She had a history of atopic dermatitis and allergy to buckwheat, raw egg and latex. Two months previously she had developed whole body flushing during dental treatment, and latex glove used by the dentist had been suspected as the cause. Prior to the operation she underwent internal examination and intrauterine echogram in which a latex glove was carelessly used by another gynecologist who had not confirmed her past history. After 30 minutes, dyspnea and urticaria without itching, appeared suddenly. Blood pressure decreased to 80/50 mmHg and heart rate increased to 120 beats.min-1. She was then transferred to our ICU. Methylprednisolone was administered intravenously for dyspnea and circulatory collapse. After 3 hours, the patient made an uneventful recovery. The increased plasma latex protein-specific IgE levels confirmed anaphylaxis to latex. The increasing incidence of potentially life-threatening allergic reactions to latex has caused mounting concern over recent years. We may suspect latex allergy when an anaphylaxic reaction or shock of unknown origin occurs. In hospitals, latex free products must be prepared for use with latex allergic patients and for protection of medical staff with this allergy. PMID- 11025963 TI - [Anesthesia for a cervical myelopathy patient with tracheal stenosis]. AB - A 71-year-old male with cervical myelopathy was scheduled for C 3-7 laminectomy. Once he had been rejected of general anesthesia in other hospital because of his tracheal stenosis. The diameter of the narrowest part of his trachea was 5 mm probably resulting from tracheostomy at 2 years of age. His cervical myelopathy seemed to be no problem for anesthesia induction because he had no problems in his neck movement and opening mouth. We evaluated his tracheal stenosis carefully using bronchofiberscopy, chest X-ray, computed tomography and pulmonary function tests. After intravenous fentanyl 100 micrograms and midazolam 2.5 mg, it was impossible to ventilate the patient. Therefore, tracheal intubation was performed immediately after lidocaine administration into the trachea. During surgery, tracheal tube dilated the narrowed portion. After surgery, the tracheal tube was removed under bronchofiberscopic observation without any problems. Tracheal stenosis was observed by chest X-ray after surgery, but he had no complaints. PMID- 11025964 TI - [Complications of hysteroscopical myomectomy: a report of two cases]. AB - Hysteroscopical myomectomy has recently become popular in Japan. We present two patients who developed water intoxication and air embolism during surgery. [Case 1] Hysteroscopical myomectomy was performed under general anesthesia in a 37-yr old woman (ASA I). Three hours after the start of the surgery, the patient's serum sodium concentration dropped to 118 mEq.l-1. She was treated with furosemide and recovered without sequelae. [Case 2] A 39-yr-old woman (ASA I) was scheduled to have hysteroscopical myomectomy under spinal and epidural anesthesia. Forty-five minutes after the start of the surgery, the patient complained of severe back pain, her blood pressure decreasing to 40 mmHg, SpO2 decreased to 80%, and ECG showed atrial fibrillation. After administration of ephedrine 5 mg, she recovered within 20 min. No abnormality was observed in echocardiogram, although some negative spots were detectable in a lung scintigraphy. She was discharged without sequelae. The hysteroscopical procedure is considered a non-invasive surgery, but the cases presented here emphasize the necessity for close attention to complications, especially pulmonary embolism. PMID- 11025965 TI - [New studies on the history of anesthesiology--a new study on Seishu Hanaoka's "Nyugan Ckiken Roku" (a surgical experience with breast cancer)]. AB - Among Japanese physicians before the Edo era, Seishu Hanaoka is the most well known even in foreign countries as well as in Japan. His detailed biography is described in a monograph by Shuzo Kure published in 1923 which has been the most important book for the study of Seishu Hanaoka. Hanaoka had worked very hard in various fields as surgeons, educator, poet and community developer. However, his best noted activity was his devotion to the development of oral general anesthetic "Mafutsu-San" or "Tsusen-San". He was the first to succeed in the excision of breast cancer in a 60 year old woman named Kan Aiya under general anesthesia with this agent on Oct 13th, 1804. The details of the case have been known to us, as the manuscript on the case which is believed to be by the hand of Hanaoka is extant in the Tenri Library, Tenri University and the whole manuscripts have been printed in Kure's monograph. For the past twenty years, I have studied carefully the microfilmed manuscript and the printed sentences appeared in Kure's book to find several serious bibliographical errors and dubious points between them. They are as follows. 1) There is no definite proof that the manuscript was transcribed by Seishu Hanaoka himself. This was originally proposed by Shuzo Kure without any rational reasons. 2) There are seven fundamental and unbelievable errors of incorrect use of Chinese characters in the manuscript. These basic errors can not be committed by Hanaoka considering that he was an excellent poet. For these errors Shuzo Kure falsified them to be printed in his book. He even altered Chinese characters in one of the photographs of the manuscript in his book. 3) Shuzo Kure did not exhibit this manuscript at the exhibition on the occasion of 150 anniversary of Seishu Hanaoka's death in Tokyo, supposedly to avoid careful study by other investigators. All above mentioned findings strongly suggest us that the manuscript "Nyugan Chiken Roku" could be transcribed by one of Hanaoka's disciples and not by Hanaoka himself. PMID- 11025966 TI - [Epidemiologic trends of breast cancer]. PMID- 11025967 TI - [Breast cancer risk factors]. PMID- 11025968 TI - [Breast carcinogenesis and sex hormones]. PMID- 11025969 TI - [Oncogene and tumor suppressor gene]. PMID- 11025970 TI - [Process of the diagnosis of breast tumors]. PMID- 11025971 TI - [The diagnostic procedure for pathological discharge from the nipple]. PMID- 11025972 TI - [Physical examination of breast cancer: inspection and palpation]. PMID- 11025973 TI - [Mammography, ductography]. PMID- 11025974 TI - [Magnetic resonance imaging for diagnosis of breast cancers]. PMID- 11025975 TI - [Contrast-enhanced computed tomography for breast cancer]. PMID- 11025976 TI - [Ultrasonography for diagnosis of breast cancers]. PMID- 11025977 TI - [Positron emission tomography for diagnosis of breast cancers]. PMID- 11025978 TI - [Color Doppler ultrasonography for diagnosis of breast cancers]. PMID- 11025979 TI - [Thermography for diagnosis of breast cancers]. PMID- 11025980 TI - [Fine needle aspiration cytology(FNAC),imprint cytology of nipple discharge]. PMID- 11025981 TI - [Stereotactic breast biopsy]. PMID- 11025982 TI - [Ultrasound-guided needle biopsy of the breast]. PMID- 11025983 TI - [Mammoscopy (duct endoscopy)]. PMID- 11025984 TI - [Pathology of breast cancer]. PMID- 11025986 TI - [Borderline lesion in breasts]. PMID- 11025985 TI - [Benign diseases of breast]. PMID- 11025988 TI - [Estrogen and progesterone receptors in breast cancer]. PMID- 11025987 TI - [Tumor markers in breast cancer]. PMID- 11025989 TI - [Clinicopathological value of ER alpha, ER beta and those isoforms in breast cancer]. PMID- 11025990 TI - [Indications for mastectomy and the surgical technique]. PMID- 11025991 TI - [Operation for breast cancers]. PMID- 11025992 TI - [Radiotherapy for breast cancers]. PMID- 11025993 TI - [Endoscopic partial mastectomy for breast cancer]. PMID- 11025994 TI - [Sentinel node biopsy]. PMID- 11025995 TI - [Breast reconstruction]. PMID- 11025996 TI - [Adjuvant chemo-endocrine therapy for early breast cancer patients--review of the literature]. PMID- 11025998 TI - [Neoadjuvant chemotherapy for breast-conservative surgery]. PMID- 11025997 TI - [Neoadjuvant chemotherapy for locally advanced breast cancer]. PMID- 11025999 TI - [High-dose chemotherapy for treatment of breast cancers]. PMID- 11026000 TI - [Follow up of breast cancer patients after surgery]. PMID- 11026001 TI - [Treatment guideline for advanced or recurrent breast cancer]. PMID- 11026002 TI - [Standard chemotherapy for metastatic breast cancer]. PMID- 11026003 TI - [Docetaxel hydrate for treatment of advanced breast cancers]. PMID- 11026004 TI - [Paclitaxel for treatment of advanced breast cancers]. PMID- 11026005 TI - [Irinotecan hydrochloride for treatment of advanced breast cancers]. PMID- 11026006 TI - [Overview of vinorelbine for metastatic breast cancer]. PMID- 11026008 TI - [S-1 for treatment of breast cancers]. PMID- 11026007 TI - [Capecitabine for treatment of advanced breast cancers]. PMID- 11026009 TI - [High-dose chemotherapy for treatment of breast cancers]. PMID- 11026010 TI - [Management of adverse effects by cancer chemotherapy for treatment of advanced breast cancers]. PMID- 11026011 TI - [Antiestrogenic agents for treatment of advanced breast cancers]. PMID- 11026012 TI - [Medroxyprogesterone acetate for treatment of advanced breast cancers]. PMID- 11026013 TI - [Aromatase inhibitors for treatment of advanced breast cancers]. PMID- 11026014 TI - [LH-RH analogue for the treatment in premenopausal, receptor-positive breast cancer]. PMID- 11026015 TI - [Pure antiestrogen for treatment of advanced breast cancers]. PMID- 11026016 TI - [Trastuzumab for treatment of advanced breast cancers]. PMID- 11026017 TI - [Bisphosphonate for treatment of advanced breast cancers]. PMID- 11026018 TI - [Tumor vaccine for breast cancer]. PMID- 11026020 TI - [Gene therapy of breast cancer]. PMID- 11026019 TI - [Immunotherapy of breast cancer: identification of breast cancer antigens by SEREX]. PMID- 11026021 TI - [Radiotherapy for treatment of advanced breast cancers]. PMID- 11026022 TI - [Staging and prognosis of breast cancer]. PMID- 11026023 TI - [Pathological (classical) prognostic factors of breast cancers]. PMID- 11026025 TI - [erbB2 as a prognostic factor of breast cancers]. PMID- 11026024 TI - [DNA ploidy pattern, S-phase fraction as a prognostic factor of breast cancers]. PMID- 11026027 TI - [CD44 as a prognostic factor of breast cancers]. PMID- 11026026 TI - [p53 as a prognostic factor of breast cancers]. PMID- 11026028 TI - [Bel-2 as a prognostic factor of breast cancers]. PMID- 11026029 TI - [tPA, uPA, PAI-1 as prognostic factors of breast cancers]. PMID- 11026030 TI - [Angiogenesis as a prognostic factor of breast cancers]. PMID- 11026031 TI - [Epidermal growth factor receptor as a prognostic factor of breast cancers]. PMID- 11026032 TI - [Cell cycle regulators as prognostic factors of breast cancers]. PMID- 11026033 TI - [Thymidine phosphorylase as a prognostic factors of breast cancers]. PMID- 11026034 TI - [Micrometastasis of bone marrow as a prognostic factor of breast cancers]. PMID- 11026035 TI - [Lymph node micrometastasis as a prognostic factor of breast cancers]. PMID- 11026036 TI - [Allelic loss as prognostic factor in breast cancer]. PMID- 11026037 TI - [Recent prognostic factors of breast cancers]. PMID- 11026038 TI - [Effectiveness of breast cancer screening]. PMID- 11026039 TI - [Effectiveness of breast self-examination for breast cancer]. PMID- 11026040 TI - [Assessment of quality of life in breast cancer patients]. PMID- 11026041 TI - [Breast-conserving treatment and quality of life]. PMID- 11026042 TI - [Informed consent for treatment of primary breast cancer]. PMID- 11026043 TI - [Informed consent for the treatment of relapsed breast cancer]. PMID- 11026044 TI - [Genetic diagnosis of hereditary breast cancer]. PMID- 11026045 TI - [Clinicopathological characteristics of hereditary breast cancer]. PMID- 11026047 TI - [Day surgery for breast cancer]. PMID- 11026046 TI - [Prevention of familial breast cancers]. PMID- 11026048 TI - [Outpatient chemotherapy for breast cancers]. PMID- 11026049 TI - [Hormone replacement therapy and breast cancer]. PMID- 11026050 TI - [Oral contraceptives and breast cancer]. PMID- 11026051 TI - [Breast cancer in the elderly]. PMID- 11026053 TI - [Treatment of breast carcinoma associated with pregnancy and lactation]. PMID- 11026052 TI - [Male breast cancer]. PMID- 11026054 TI - [Chemosensitivity test of breast cancer]. PMID- 11026055 TI - [Breast cancer chemoprevention (tamoxifen, raloxifene, and others)]. PMID- 11026056 TI - [First detection of human calicivirus infection in Hungary]. AB - Human caliciviruses (HuCVs) are important pathogens all over the world. They cause acute non-bacterial gastroenteritis in humans in all age-groups. The aim of the study was to detect HuCV infection in Hungary. Stool samples examined were received from a food-borne outbreak in nursery and elementary school in Szeged and Algyo areas November 1998. Reverse transcription-polymerase chain reaction (RT-PCR) method was successfully used for the detection of HuCVs in all stool samples received. This is the first description of HuCVs in Hungary and the verification of them as causative agents in a food-borne outbreak. PMID- 11026057 TI - [Endoscopic harvesting of the radial artery at coronary artery bypass graft: a new surgical technique]. AB - The radial artery as arterial graft is applied ever increasingly in coronary artery bypass grafting. With the endoscopic technique described by the authors, the radial artery can be lifted out extremely quickly and gently through transverse 2 cm incisions, using a so-called "spiral instruments for radial artery harvesting" and a 30 degrees endoscope. The ultrasonic knife used for incision and hemostasis does not cause thermic and mechanical damage. The radial artery and accompanying veins remain protected inside the spiral interior. The authors present a case where this method was applied successfully. PMID- 11026059 TI - [A stem cell ... is stem cell?]. AB - We are used to associate stem cells with renewable tissues such as blood, gut and skin. But some cells in the adult central nervous system have the capacity to generate new neurons and glial cells as well and as such, they are considered to be neural stem cell. Yet their ability to generate neurons and glia, and their presence in the central nervous system throughout life, suggests new, intriguing possibilities for recovery and repair after damage to the central nervous system- and unexpectedly, the regeneration of blood tissues. After transplantation into irradiated hosts, neural stem cells were found to produce a variety of blood cell types including myeloid and lymphoid cells as well as early hematopoietic cells. Therefore, the developmental potential of stem cells is not restricted to the differentiated elements of the tissue in which they reside. Multipotential stem cells can persist in an undifferentiated state, and depending on specific environmental conditions function as a stem cell for many different tissues. PMID- 11026058 TI - [Double flap reconstruction of extended facial defects]. AB - The authors report the reconstruction of large facial defects remaining after resection of two advanced facial cancer cases. In both cases double flap closures were carried out. In the first case the internal flap was created from the remaining part of the oral mucosa membrane, while in the second a forearm fascio cutan free flap was used. For external closure in both cases platysma based transpositional flaps were used, prepared from the neck. On the basis of excellent aesthetic results and safe applicability the authors recommend the more frequent use of this method. PMID- 11026060 TI - [Sjogren's syndrome and multiple myeloma]. AB - In our recent work we present two cases of coexisting Sjogren's syndrome and myeloma multiplex. Our results obtained studying a high number of patients suggest that the occurrence of myeloma in Sjogren's syndrome is much higher than in normal population. This can possibly be explained by the common pathogenetical factors of the two diseases. The survival of one of our two patients in 4 years, and her disease is still in complete remission. The possible explanation of the therapeutical success can be the rather aggressive chemotherapeutical approach in her case. Perhaps the prognosis of myeloma coexisting with Sjogren's syndrome is different that of pure myeloma, it is suggested that the earliest possible aggressive therapy of myeloma accompanied by Sjogren's syndrome can be useful and recommendable. PMID- 11026061 TI - [Successor of Vilmos Tauffer at the gynecology department chair: Istvan Toth (1865-1935)] . PMID- 11026062 TI - [Therapeutic herbs in ancient Chinese medicine]. PMID- 11026063 TI - [Remembering Prof. Dr. Janos Batizfalvy (1895-1960) on the 40th anniversary of his death]. PMID- 11026064 TI - [Comments on the article "Evolution of air contamination and impact of control programs in 3 megacities in Latin America"]. PMID- 11026065 TI - [Child abuse: a world problem in public health]. PMID- 11026066 TI - [Comments on the article "Experience in the management of the HIV-patient among physicians of the Health Department"]. PMID- 11026067 TI - [Agreements and disagreements with comments by Dr. Saavedra]. PMID- 11026068 TI - [Argument claiming the possible regression to the media in a study on educational intervention in women to detect breast cancer]. PMID- 11026069 TI - [Levels of plasmatic lead in children 8-10 years of age and its relation to changes in visual-motor system and balance]. AB - OBJECTIVE: To assess the association between blood lead concentrations and visual motor coordination and equilibrium in school age children. MATERIAL AND METHODS: In November-December 1998, a cross-sectional study was conducted among 255 children aged 8-10, who attended public schools in Sector 1 of the Oaxaca State Public Education Institute. Data were collected using the Frostig Evaluation of Visual Perception test and the equilibrium subscale of the Frostig Movement Skills Test Battery. A blood sample was taken to measure lead levels by atomic absorption spectrometry. Socioeconomic data and health histories were collected for use as control variables. Statistical analysis consisted of multiple regression models to test the relationship between blood lead level and the visual-motor and equilibrium tests. We assessed the effect of lead within the model using 1,000 Montecarlo simulations. RESULTS: The geometric mean of blood lead concentrations was 11.5 micrograms/dl (geometric standard deviation +6.3, 5.2). After adjusting for control variables, the visual-motor integration subscale was significantly related to blood lead concentration (p > 0.042). The visual-motor integration value decreased 1.78 (95% CI -3.51, -0.06) points for each 10 micrograms/dl increase in blood lead concentration. Among the four sub tests comprising the visual-motor integration subscale, only eye-hand coordination (p = 0.045) and spatial relations (p = 0.039) were significantly related to blood lead. The visual-motor integration subscale was also significantly related to family income; greater income was related to greater testing scores. Only 3.1% of the children had clinically abnormal testing scores. No statistically significant association was found between blood lead levels and the Frostig subscale Index of Reduced Motor Response, the General Visual Perception Index, or any other equilibrium tests. CONCLUSIONS: The inverse relationship between blood lead concentration and visual-motor skill is consistent with results from studies in other countries. Blood lead levels common among children of Oaxaca are sufficient to produce subtle visual-motor impairments. These findings point out the need to strengthen the initiative to reduce child exposure to known lead sources, particularly lead-glazed ceramics, and thus lower the blood lead levels of the population. PMID- 11026070 TI - [Relationship between emergency consultations for respiratory diseases and air pollution in Juarez City, Chihuahua]. AB - OBJECTIVE: To assess the relationship of < or = 10 microns particles (PM10) and atmospheric ozone concentrations, with the daily number of emergency visits due to asthma and acute respiratory diseases, among children aged under 15, living in Ciudad Juarez, Chihuahua, Mexico. MATERIAL AND METHODS: Between 1998 and 1999, an ecologic study was conducted. Atmospheric data were obtained from the Environmental Protection Agency (EPA), from eight monitoring stations located in Ciudad Juarez, Chihuahua, and EI Paso, Texas. From July 1997 to December 1998, data from emergency room visits for respiratory illness were abstracted from existing medical records of two Mexican Institute of Social Security (IMSS) hospitals in Ciudad Juarez. Diagnoses were classified into two groups: a) asthma, and b) upper respiratory infections (URI), according to the International Classification of Diseases (ICD-9 and/or IDC-10). Statistical analysis was carried out using the Poisson regression time series method. RESULTS: During the study period, the mean 24-hour PM10 level was 34.46 micrograms/m3 (SD = 17.99) and the mean ozone level was 51.60 ppb (SD = 20.70). The model shows that an increase of 20 micrograms/m3 in the mean 24-hour exposure to PM10 was related to an increase of 4.97% (95% CI 0.97-9.13) in emergency visits for asthma, with a 5 day lag, as well as to an increase of 9% (95% CI 1.8-16.8) when a cumulative 5 day exposure was considered. URI increased 2.95% as a cause of emergency room visits, for each 20 micrograms/m3 increase in the mean 24-hour exposure to PM10. The impact of PM10 on emergency visits for asthma was greater on days with ozone ambient levels exceeded 49 ppb (median value). CONCLUSIONS: A positive association was found between environmental PM10 and ozone concentrations and the daily number of emergency room visits due to asthma and acute respiratory diseases, even with levels lower than the Mexican standard levels. Also, a synergic effect between PM10 and O3 was found. PMID- 11026071 TI - [Availability of essential medications at primary level units of the Health Department of Tamaulipas, Mexico]. AB - OBJECTIVE: To describe the availability of some essential drugs at the primary health care units of the Ministry of Health of Tamaulipas, Mexico. MATERIAL AND METHODS: Between September and October 1998, all first level healthcare units of Tamaulipas' three sanitary jurisdictions were surveyed. Drug availability was assessed. The measurement instrument was a checklist of 56 drugs and 10 different supplies. For each drug and input the absolute number and the proportion of units with this drug or input was calculated. In the units where the drugs were available, the medians were calculated. The median of the total number of drugs available in all units was used as a global indicator. This same exercise was developed for each unit. Comparisons between the availability of these inputs in the units and stockrooms were also done. Stata 5.0 was used for statistical analysis. RESULTS: None of the inspected units had full availability of all checklist drugs. The highest percentage of drug availability was 84% and the lowest was 32%. There was limited availability of antibiotics, antihypertensive, hypoglycemic, and iron deficiency drugs. The availability of oral rehydration salts and contraceptive and vaccine agents was acceptable. CONCLUSIONS: Healthcare organizations must find alternative ways to improve access to drugs nationwide, in general, and availability of essential drugs in first level healthcare units, in particular. Two recent initiatives provide an optimistic outlook: decentralization of health services for the uninsured and the Generic Exchangeable Drugs Program, established nationwide in 1998. PMID- 11026072 TI - [Effect of budesonide on bone density and metabolism in asthmatic children]. AB - OBJECTIVE: To assess the effect of inhaled budesonide on the mineral density, content and bone metabolism in children with asthma. MATERIAL AND METHODS: From September 1996 to July 1997, a cross-sectional study was conducted in 38 prepubertal children aged 6 to 11 years, selected from the pediatric chest outpatient clinic of the Instituto Autonomo Hospital Universitario de Los Andes, Merida, Venezuela. Three study groups were assembled: 9 asthmatic children treated with inhaled budesonide (300 micrograms/day) for over 6 months (Group A); 14 asthmatic children not treated with inhaled corticosteroids (Group B); and 15 non-asthmatic children (Group C). All of them underwent testing of bone formation and resorption markers, and measurement of bone mineral density (DMO) and content (CMO). Statistical analysis consisted of central tendency and dispersion measures, analysis of variance, and Fisher and Scheffe tests for comparison of means. RESULTS: In the groups studied (A, B, and C) calcium serum levels were 9.1 +/- 0.3; 9.6 +/- 0.4; 9.3 +/- 0.6 mg/ml, respectively; osteocalcin levels were 14.8 +/- 4.6; 13.0 +/- 2.5; 11.9 +/- 3.4 ng/dl; the type I collagen carboxyterminal telopeptide (ICTP) levels were 19.6 +/- 16.5; 14.2 +/- 15.4; 13.0 +/- 18.3 micrograms/l; the DMO levels were 0.67 +/- 0.06; 0.68 +/- 0.06; 0.69 +/- 0.06 g/cm2; and the CMO levels were 1,158.8 +/- 217.4; 1,106.4 +/- 256.1; 1,176.5 +/- 240.5 g, respectively. No statistically significant differences were observed between the groups. CONCLUSIONS: The administration of 100-400 micrograms/day of inhaled budesonide for a period of six months, did not change the bone mineral density and metabolism of asthmatic children. PMID- 11026073 TI - [Validity and reproducibility of a questionnaire on physical activity and non activity for school children in Mexico City]. AB - OBJECTIVE: To assess the validity and reproducibility of a self-reported questionnaire on physical activity and inactivity, developed for children aged 10 14 in Mexico City. MATERIAL AND METHODS: Between May and December 1996, a self reported physical activity and inactivity questionnaire was developed and applied twice to a sample of 114 students aged 10 to 14, from a low and middle income population of Mexico City. The children's mothers completed the same questionnaire, and two 24-hour recalls of physical activity were used for comparison. Statistical analysis consisted of central tendency and dispersion measures and Pearson's correlation coefficient. RESULTS: Correlations between hours per day spent in physical activity and inactivity from the children's questionnaire and the 24-hour recall data, were 0.03 for moderate activity, 0.15 for vigorous activity, and 0.51 (p = 0.001) for watching television, adjusted by age, gender, town, and illness prior to the administration of the questionnaire. Compared to the 24-hour recall data, the questionnaire overestimated the time spent watching television, reading or participating in vigorous activity, and underestimated the time engaged in moderate activity. Statistically significant (p < 0.05) six-month reproducibility values were observed for watching television (r = 0.53), sleeping (r = 0.40), moderate (r = 0.38), and vigorous activity (r = 0.55). CONCLUSIONS: Among children of Mexico City aged 10-14, the questionnaire showed acceptable validity in estimating the time watching television, and acceptable reproducibility of the time watching television, vigorous and moderate activity. PMID- 11026074 TI - [Epidemiologic monitoring system for the diabetic patient; use of computerized technology in the quality of medical care]. AB - OBJECTIVE: To describe an instrument for epidemiologic surveillance of diabetes mellitus and evaluating the quality of care in primary health care. MATERIAL AND METHODS: The study was conducted from January 1, 1998, to June 30, 1999, at the Epidemiology and Health Services Research Unit of the Mexican Institute for Social Security (IMSS), in Hermosillo, Sonora. A single data collection form was designed, which contains items included in the diabetes care clinic of the Unit of Family Medicine of the Mexican Institute of Social Security. RESULTS: We developed and implemented a software program to enter data from the paper format and to generate individual and group reports on attendance to scheduled medical visits, medical history, evolution of signs and symptoms, laboratory tests, and medication. CONCLUSIONS: Electronic data systems allow the availability of reliable and continuing information for surveillance of the comprehensive care of the diabetic patient. PMID- 11026075 TI - [Changing pattern and frequency of lung cancer in Mexico]. AB - OBJECTIVE: To assess the frequency, histological type and the epidemiological pattern of lung cancer (LC), during a 40 year period, at the National Institute of Respiratory Diseases (INER). MATERIAL AND METHODS: This cross-sectional study was conducted in 1999. Data were abstracted from the INER's service reports for 1983-1996 and the surgical pathology archives for 1957-1996. Data from 1961 patients were analyzed, to obtain frequency measures on: the characteristics of INER LC patient admissions, age, sex, histological studies, specimens and diagnoses. RESULTS: Between 1983-1996, LC ranked eighth to second place in INER inpatient admissions. Results were grouped by decades. In the first two decades (1957-1976), 9% of patients were female; 32% in the third (1977-1986), and 38% in the last decade (1987-1996). Epidermoid cell carcinoma was more frequent in the two first decades (61%) and adenocarcinoma in the last two, (41% and 62%, respectively.) CONCLUSIONS: LC is currently more frequent in women, and adenocarcinoma is the most frequent neoplasia. PMID- 11026076 TI - [Main measurements in epidemiology]. PMID- 11026077 TI - [Experimental clinical studies]. PMID- 11026078 TI - [A clinical study of psychopathology in systemic lupus erythematosus]. AB - Thirty patients (24 inpatients and 6 outpatients) with a clinical diagnosis of SLE were examined between September 1, 1998 and August 1, 1999 in the rheumatology clinic of Jichi Medical School Hospital. All of these patients fulfilled the 1982 revised criteria of the American Rheumatism Association for the classification of SLE and had some psychiatric manifestations (psychiatric SLE; P-SLE group). Mean patient age was 38.6 +/- 13.0, and there were 5 males and 25 females. When classified into 5 subgroups according to the most prominent symptoms, the distribution was as follows: consciousness disturbance group: 6 (20%), schizophrenia-like group: 5 (16.7%), mood disorder group: 7 (23.3%), neurosis-like group: 10 (33.3%), and convulsive disorder group: 2 (6.7%). Among all 37 psychiatric episodes, symptoms appeared in 37.8% of cases during the acute phase of SLE (during onset or recurrence) and in 62.9% during the chronic phase (during remission). The profile of the P-SLE group showed that the psychiatric symptoms of the SLE patients were milder and more chronic than those described in previous reports. To begin to comprehend the psychopathology of SLE, we put forward the concept of "Psychiatric basal state" and "psychiatric conjugated state". The former is considered a direct reflection of the acute-phase SLE process on mental condition. It is defined clinically as psychiatric symptoms that parallel the activity of SLE and respond well to steroid therapy. The latter include all other psychiatric problems in which one cannot rule out the effects of pharmacological, somatic, personality, and environmental effects on psychiatric symptoms. Only 3 patients in the P-SLE group fulfilled the criteria for the "psychiatric basal state". All three patients belonged to the consciousness disturbance group, whose clinical features were defined as slight clouding of consciousness, so-called "Amentia" in the sense of the German terminology. The clinical profile of this state is: 1. the patients are young (about 16 years old), 2. the onset of psychiatric symptoms is within 5 years after the onset of SLE, 3. confusion and disorientation are the most characteristic features, and 4. the clinical course of this state is almost 2 months. The experience structure of the "psychiatric basal state" consists of: 1. difficulty in selecting and holding a topic in cognition, 2. confusion and emotional instability as the basal mood, and 3. primitive and floating forms of delusions and hallucinations. Using this concept of the "psychiatric basal state" as a clue, we can hypothesize the continuity of diverse psychiatric symptoms in SLE. The "proper process of SLE (Harada)" has a disintegrating effect on the "ego" and it allows various psychopathological phenomena to emerge in the experience field. Against this background, additional factors, such as secondary organ damage, personality structure, and social environment, induce organization of the "psychiatric conjugated state". PMID- 11026079 TI - [A survey of drug utilization in psychiatric hospitals in Japan: comparison of 1973, 1979, and 1993]. AB - This paper investigated the trend in prescription patterns in the treatment of psychiatric inpatients using a database of our 1993 survey as well as reports of surveys conducted by Ito et al. in 1973 and 1979. To make the database of our survey comparable with surveys in 1973 and 1979, we extracted and analyzed 1,164 cases which met the criteria of Ito's survey, requiring that patients were diagnosed as having schizophrenia and were hospitalized for two years or longer. The results were as follows: 1) The most common number of different drugs per patient increased from 4 in 1973 and 1979 to 8 in 1993. That of psychotropics increased from 2 to 5. That of neuroleptics increased from 2 to 3. 2) The most commonly prescribed neuroleptics were haloperidol followed by levomepromazine and chlorpromazine. While the top three neuroleptics had not changed, the mean daily dose of haloperidol rose from 8.4 mg in 1979 to 15.3 mg in 1993. PMID- 11026080 TI - [Better overweight and fit. Interview by Dr. Klaus Reinhardt]. PMID- 11026081 TI - Obesity management--new perspectives. AB - Obesity represents a growing threat for the health of population worldwide. The decline of physical activity and the passive overconsumption of energy-dense, high-fat diets are important factors that explain the increased prevalence of obesity. Despite many efforts, the prevention and the treatment of obesity are often a failure. Since obesity development is due to a chronic imbalance between energy intake and energy expenditure, the most important advice for the prevention and the treatment of this nutritional disorder remains the reduction of high-fat foods and the stimulation of fat oxidation by promoting physical activity. The recent advances in our understanding of the control of food intake and of energy expenditure offer the hope that new therapeutic agents will become available over the next decade. This article briefly presents our present understanding of the main mechanisms which are responsible for the increased prevalence of obesity and some new areas of research which may be promising for the prevention and treatment of this disease. PMID- 11026082 TI - [Epidemiology of obesity]. AB - Obesity is a well-recognized health hazard in affluent societies. The prevalence of overweight and obesity are high and still increasing consistently in industrialized countries. The body mass index (BMI) is an internationally accepted index to estimate body weight. The recent published WHO-classification of body weight allows the worldwide comparison of obesity prevalences. The WHO MONICA study revealed different obesity prevalences in several European countries. The lowest prevalence of obesity was found in Sweden and Northern France (7-14%) whereas the highest prevalence (40%) was reported from East European countries. The results of the DHP-survey suggest that 50% of the adult German population are overweight and 20% are obese. Data from the MONICA-study show lower prevalences for Switzerland, where 38% of the adult population are considered to have overweight and 11% are obese. By the way the prevalence of obesity is still increasing with a renewed acceleration during the last two decades. Obesity is starting to get the status of a main health problem not only in western countries but also in less developed countries of Middle- and South America. In Germany obesity and the obesity-related morbidity and mortality caused costs of nearly 20.7 Mrd. DM in 1995. Therefore obesity is a major economic burden for the society. Like in other developed countries the costs attributable to obesity correspond to 2-8% of total health care expenditure. PMID- 11026083 TI - [Etiology of overweight and obesity]. AB - Obesity results from a predominance of caloric intake over energy expenditure. Twin, adoption and family studies have demonstrated that, together with environmental conditions, various genetic factors play an important role in the pathogenesis of obesity. In recent years, it was possible to identify several defects in single genes responsible for obesity in rodents, some of which may also be involved in human obesity. Besides leptin as the most notable example, numerous other proteins and neuropeptides have been identified in recent years that participate in a complex network to regulate food intake and energy expenditure. The ongoing search for the important obesity genes should not only result in a better understanding of energy metabolism, but may also help in the development of new strategies for the treatment of obese patients. PMID- 11026084 TI - [Adipose tissue and obesity]. AB - Adipose tissue is not simply a storage depot. Adipocytes secrete hormones, growth factors and cytokines, such as leptin and TNF-alpha, as well as proteins that are related to the immune system and vascular functions. Through this network of endocrine, paracrine, and autocrine signals fat cells participate in the regulation of energy homeostasis, host defense and reproduction, and may also contribute to the development of pathological states, such as insulin resistance. Adipose tissue is confined to distinct depots. In Cushing's disease or following treatment of AIDS, certain adipose depots enlarge whereas others shrink, suggesting the existence of site-specific differences in fat cell function. Increases in adipocyte number occur via replication of preadipocytes, a process that is not restricted to infancy but occurs throughout life. In contrast to still widely-held beliefs, mature fat cells can be eliminated by dedifferentiation or apoptosis. PPAR-gamma, a transcription factor that is activated by fatty acids and prostaglandins, plays a central role in adipose conversion of preadipocytes and appears to participate in controlling the size of mature fat cells as well. PMID- 11026085 TI - [Consequences and complications of obesity]. AB - Obesity increases the risk of metabolic complications such as diabetes, dyslipidemia, systemic hypertension and cardiovascular disease. These are mainly responsible for the increased mortality of obese people. Other metabolic consequences of obesity are: gallstones, steatosis of the liver and the polycystic ovary syndrome. Beside the body mass index the distribution of body fat is important. Centralized obesity, as measured by the waist-to-hip circumference ratio (WHR), is associated with increased mortality and morbidity. Insulin resistance and hyperinsulinaemia seem to play a central role in the pathogenesis of this association. Obesity has not only metabolic complications. There is a relationship between obesity and impaired respiratory function. Furthermore is obesity a risk factor for osteoarthrosis of the knee, the hip and even the hand and for pulmonary embolism and venous thrombosis. Obesity can also lead to psycho-social problems such as depression, social discrimination and isolation. PMID- 11026086 TI - [Obesity clinic]. AB - The cornerstone of the evaluation of an obese patient is the medical examination in combination with a few selected obesity specific measurements. Key elements in the obesity specific history are the patient's weight history, the diet history, evaluation of the present and past physical activity pattern and the evaluation of the patient's target weight. Central elements in the examination are the computation of the body mass index (BMI) as well the measurement of the waist circumference. The waist circumference shows a higher degree of correlation with different morbidities than the BMI. A waist circumference of > 80 cm in women and > 94 cm in men is associated with an increased overall morbidity risk. In general a minimal biochemical work-up--including fasting glucose, total cholesterol, HDL and triacylglycerol, urate, electrolytes and TSH--is enough. Special tests (screening examination for e.g. M. Cushing) are only indicated in the case of clinical suspicion; the determination of leptin is presently of no diagnostic nor therapeutic relevance. The indication for weight reduction should be formulated individually. In the long term weight stability has to be regarded as a success for most patients. Presently the prevention of weight gain and obesity is still the safest and most efficient "therapeutic" approach. PMID- 11026087 TI - [Eating behavior, eating disorders and obesity]. AB - Over the last 50 years, the nutritional and socioeconomic conditions have dramatically changed in all industrialized countries. As a consequence, there has been a sharp rise in the prevalence of obesity. Simultaneously, social and cultural pressures to maintain a thin body shape have significantly increased. This untoward situation is largely responsible for the steady increase of eating disorders, especially bulimia nervosa and binge-eating disorder, which are common disorders among normal or overweight individuals. Although the criteria for bulimia nervosa were first described in the DSM-III in 1980 (APA, 1980), recent studies have demonstrated that only about 12% of these patients are detected by their GP's. One reason for this low rate of detection may be due to the tendency of patients to conceal their illness from others. It is also possible, however, that general practitioners lack sufficient knowledge about bulimia nervosa, preventing proper identification. To help improve this situation, diagnostic guidelines and therapeutic options were summarized. Binge-eating disorder (BED), which is classified as an "eating disorder not otherwise specified" in the DSM-IV (APA, 1994), has been described as the most relevant eating disorder for overweight individuals. It has been estimated that approximately 20-30% of overweight persons seeking help at weight loss programs are classified as binge eaters. Initial results from these studies suggest that binge eaters may require a modified psychotherapeutic approach which focuses on normalizing disordered eating patterns before attempting weight loss. In addition to the importance of screening for eating disorder behaviors, overweight patients should be assessed for other comorbid conditions, such as depression and anxiety. Further, body image disturbances should be assessed during the evaluation. In the event that comorbid disorders are present, it is recommended that specific psychotherapeutic interventions which target these problems be integrated into the overall weight reduction program. PMID- 11026088 TI - [Principles of dietary treatment of obesity]. AB - One of the first steps in a clinical approach to any obese subject should be focused on the reduction and/or normalization of any potential or existing metabolic abnormality. Overeating and/or unbalanced food intake remains the major element in the origin and maintenance of obesity. The reduction of energy intake is the basis of successful weight loss. In obese subjects there are huge amounts of energy stored, mainly in the adipose tissue, which are mobilized according to the size and duration of an energy deficit. Considerable studies have been devoted to finding the optimal dietary approach that would promote rapid weight loss while maximizing the depletion of adipose tissue and conserving body protein. During fasting adipose tissue lipolysis rate increases and liberated unesterified fatty acids are oxidized in muscle and liver. The liver produces ketones which are oxidized in muscle and brain. The energy need of the brain is not sufficiently covered by ketone oxidation, therefore additional glucose must be provided. The liver produces glucose by gluconeogenesis using amino acids from muscle protein. Because of limited protein sources, protein must be given during energy restricted diet. Besides protein also vitamins, minerals, trace elements, fiber, and linoleic acid must be substituted during fasting and during treatment with very low calorie diets. Meal replacements are helpful to fulfil all the requirements. There is consensus that the first step in dietary treatment is an energy restricted diet with a calorie deficit of at least 600 Kcal/day, but more than 800 Kcal/day must be provided, with all essential nutrients. Observing the regulations, weight reduction with appropriate diet plans improves metabolic disturbances. PMID- 11026089 TI - [Weight reduction through physical exercise and diet--how much is enough and is too much possible?]. AB - Presently available literature reveals that a combination of approach has the potential to induce a body weight loss and favor weight maintenance in a reduced obese state while also normalizing the metabolic profile of these individuals. Further weight loss is thus no longer justified and might be harmful to weight maintenance and health although these individuals could still be diagnosed as obese according to actual classifying criteria for defining obesity. PMID- 11026090 TI - [Obesity: principles of drug therapy]. AB - Obesity is a major global public health problem. In many instances, a combination of diet modification, increased physical activity and behavior therapy fail or are insufficient for sustained weight loss. In these situations, drug therapy may be helpful. However, drug treatment of obesity resulted in unexpected devastating events in recent years. In the late sixties, aminorex caused an epidemic of pulmonary hypertension with high mortality rates. Dexfenfluramine and phentermine were also associated with the development of pulmonary hypertension and with alarming reports of cardiac valvular abnormalities. Therefore, these drugs were withdrawn from the market. Newer drugs, like sibutramine, a serotonin and norepinephrine reuptake inhibitor, and orlistat, a specific lipase inhibitor, reduce body weight significantly compared to placebo. In combination with a hypocaloric diet, weight loss of three to ten kilos can be achieved. Pharmacotherapy is limited to patients with a body mass index greater than 30 kg/m2, if non-pharmacological treatment programs have failed. The drugs should be prescribed under strict medical surveillance only. PMID- 11026091 TI - [Principles of surgical treatment of morbid obesity]. AB - The incidence of morbid obesity has steadily increased over the last few years. Although conservative treatment of morbidly obese patients made some progress (e.g. Xenical), a persistent weight reduction can hardly be achieved in these patients. The possibility of a minimal invasive approach lead to a revival of the surgical therapy of morbid adiposity. The laparoscopic gastric banding became an attractive therapy for patients presenting with a body mass index greater than 40 kg/m2. However, the long-term follow-up after laparoscopic gastric banding increasingly shows the limit of this surgical approach and emphasises the importance of a determined preoperative patient selection. This article reviews the clinical basics and functional mechanisms of different surgical methods to treat morbid obesity comparing their advantages and disadvantages. Due to the widely and nowadays very popular application of laparoscopic gastric banding, this operative method is described in detail. PMID- 11026092 TI - [Obesity in children and adolescents]. AB - The prevalence of obesity in increasing worldwide, and its complications are now better recognized. As childhood obesity tracks into adulthood, the treatment and prevention of this condition is necessary early in life. This review describes the public health impact of pediatric obesity, its risk factors, and suggestions for its treatment and prevention, including behavior modifications, decrease in inactivity, increase in physical activity, and dietary changes. PMID- 11026093 TI - [Contraindications to weight reduction]. AB - It is relatively well accepted that weight gain, even within the range of normal weight, is detrimental for health. The claimed long-term benefit of intentional weight loss is mainly based on a few observational trials, confounding intentional and non-intentional weight loss. The few data on obesity-related diseases prevented by intentional weight reduction have not been replicated. Thus, for lack of level-one evidence it is to date doubtful whether voluntary weight loss should be intensively recommended to obese individuals rather than other lifestyle-interventions, e.g. exercise training. The aim of this overview is to discuss some of the contraindications to intentional weight loss often ignored in recent debates. There is no intention to question the increased risk of overweight on morbidity and the need for preventing weight gain in our population. Besides well-known consequences secondary to rapid weight loss, e.g. gallstones and electrolyte disorders, some new aspects or more debated issues are discussed. Recent compelling data indicate a significant bone density loss after weight loss of a few kilograms. Knowledge on the impact of weight cycling mainly relies on cross-sectional data. So far there is no concluding evidence of adverse pathophysiological effects from weight cycling. Repeated dieting has been associated with eating disorders, although the cause-effect relationship has not been well established. At least on an empirical basis there seems to be a tolerance effect after repeated weight losing efforts. Pharmacotherapy tends more and more to be part of weight loss interventions. Although the currently available drugs are designed for long-term treatment, in practice they are rarely used longer than a few months, and therefore their use can be entirely questioned. Nonetheless, if employed properly for weight loss and weight maintenance, i.e. "for life", additional data on long-term effects on health are needed. Such risks can then be weighed against the risk of remaining obese. Even a limited detrimental effect on blood pressure, such as known for sibutramine, could compromise the beneficial effect of the drug on weight loss and maintenance. Further, the financial burden of weight loss is not to be underestimated. Besides diets and weight loss programs being a multi billion dollar business, the cost of patient care in cases of surgery for gallstones has, among other economic implications, to be considered. Finally, with the example of the recently investigated impact of fitness level on health, exercise training may be an alternative to weight loss. Whether obese or not, cardiovascular fitness is a strong, independent predictor of cardiovascular mortality and overall mortality. Most obese being unfit, a greater preventive impact might come from becoming more fit than from losing weight. This should only illustrate that there is still a long way to go until we have sufficient scientific data to be able to tell whether in the management of obesity weight loss or other interventions are of higher priority. In part, current recommendations to lose weight are biased by societal pressure, which overrules simple scientific evidence. PMID- 11026094 TI - Marginal zone B cells, monocytoid B cells, and the follicular microenvironment. Determinants of morphologic features in a subset of low-grade B-cell lymphomas. PMID- 11026095 TI - How specific is cardiac troponin? PMID- 11026096 TI - Recommendations for the reporting of resected esophageal carcinomas. Association of directors of anatomic and Surgical Pathology. PMID- 11026097 TI - Lymphomas with follicular and monocytoid B-cell components. Evidence for a common clonal origin from follicle center cells. AB - We investigated the clonal relationship between follicular center cell and monocytoid B-cell components of non-Hodgkin lymphoma by isolating the components and comparing the nucleotide sequences of the complementarity-determining region (CDR)3 of the rearranged immunoglobulin heavy chain (IgH) gene. Paraffin blocks from 4 cases with amplifiable DNA using the polymerase chain reaction (PCR) were identified. Multiple representative cell clusters of the 2 components were obtained by microdissection, and the IgH CDR3 was amplified using a seminested PCR. Most of the PCR products obtained from both tumor components in each case had identical lengths when analyzed with polyacrylamide gel electrophoresis (PAGE) and identical migratory patterns on denaturing gradient gel electrophoresis (DGGE). These findings indicate sequence identity of the IgH CDR3 of both tumor components. Sequence analysis showed that point mutations were responsible for bands from the same case that had nonidentical migratory patterns by DGGE. The components in each of the 4 cases studied have the same clonal origin. Intraclonal sequence variations in the IgH gene were observed in 2 cases, consistent with the presence of continued somatic hypermutation after establishment of the clone. The expression of CD10 and bcl-2, as well as the detection of bcl-2 rearrangements in 2 cases, indicate that these lymphomas are of follicular center cell origin. PMID- 11026098 TI - De novo CD5+ diffuse large B-cell lymphomas. A heterogeneous group containing an unusual form of splenic lymphoma. AB - We reviewed our institutional experience with de novo CD5+, large B-cell lymphomas to determine whether they represent a distinct entity and are related to CD5+ small B-cell disorders. We identified 13 cases with multiparameter flow cytometry over a period of 58 months (5% of large B-cell lymphomas) in 7 females and 6 males. Three groups were identified. Group 1 (2 cases) had diffuse splenic red pulp involvement with a distinctive cordal pattern of infiltration, no other clinical evidence of mass disease, microscopic disseminated disease on further workup, and an identical immunoglobulin-negative immunophenotype. Group 2 cases (7 cases) were clinically and morphologically heterogeneous and had an immunophenotype resembling mantle cell lymphoma (FMC7-positive, CD23-). Group 3 (4 cases) had miscellaneous immunophenotypes, including one closely resembling chronic lymphocytic leukemia. Cyclin D1 was positive in only 1 of 10 evaluable cases (group 2). We conclude that CD5+ diffuse large B-cell lymphomas are heterogeneous; most cases do not seem to be related to chronic lymphocytic leukemia or mantle cell lymphoma. However, we identified a subgroup of primary splenic CD5+ large B-cell lymphoma with diffuse red pulp involvement and believe this may represent a distinct clinicopathologic entity. PMID- 11026099 TI - Cyclosporine metabolism in patients after kidney, bone marrow, heart-lung, and liver transplantation in the early and late posttransplant periods. AB - Cyclosporine is used in the prevention of allograft rejection. Owing to its narrow therapeutic index, regular monitoring of the whole blood levels of cyclosporine is required. We observed that immunoassays measured significantly higher cyclosporine levels than did high-performance liquid chromatography (HPLC) over time after transplantation. As cyclosporine metabolites cross-react even with immunoassays, this observation might be due to alterations of the cyclosporine metabolism. We analyzed cyclosporine metabolite concentrations in the early and in the late posttransplantation periods in 127 patients after kidney, bone marrow, heart-lung, and liver transplantation by HPLC and determined whole blood levels of cyclosporine by 4 immunoassays (enzyme-multiplied immunoassay [EMIT], cloned enzyme donor immunoassay [CEDIA], AxSYM [Abbott Laboratories, Chicago, IL], and TDx [Abbott Laboratories]). Despite reduced dose, we found significantly higher cyclosporine concentrations measured by the EMIT, AxSYM, and TDx assays in various patient groups. These results are due to the increased metabolite/cyclosporine ratio in the late posttransplantation period. In particular, the metabolites AM1 and AM19 increased significantly over time in bone marrow transplant recipients. Therefore, cyclosporine levels measured by immunoassays should be interpreted with caution. PMID- 11026100 TI - Expression of messenger RNA of the cardiac isoforms of troponin T and I in myopathic skeletal muscle. AB - In the absence of clinical signs, elevated values of the cardiac isoforms of troponin T (cTnT) and I (cTnI) can be found in the serum samples of some patients with skeletal muscle myopathies; the cause is unclear. We studied the messenger RNA (mRNA) expression of cTnT and cTnI in the skeletal muscles of 24 patients with histologically proven myopathies and in 18 patients in whom a myopathy could be excluded. For cTnT- and cTnI-mRNA determination, we designed specific primer pairs for nested polymerase chain reaction. After amplification, the products were digested with 2 restriction enzymes and visualized. We found cTnT mRNA in 7 skeletal muscle biopsy specimens (6 patients with Duchenne muscular dystrophy, 1 patient with a primary sarcoglycanopathy) and cTnI mRNA in 6 (5 with Duchenne muscular dystrophy, 1 patient with a histologically negative biopsy). The mRNA of the cardiac isoforms, cTnT and cTnI, is expressed in the skeletal muscles of patients with Duchenne muscular dystrophy, but also in some other myopathies. Further studies are needed to show whether the mRNA is translated into the protein, but serum levels of cTnT and cTnI in patients with Duchenne muscular dystrophy would seem to indicate this. PMID- 11026101 TI - Endoscopic mapping and surrogate markers for better surveillance in Barrett esophagus. A study of 700 biopsy specimens. AB - Surveillance methods in Barrett esophagus (BE) using light microscopic examination of random biopsy specimens may miss focal dysplasia. In addition, dysplastic foci identified initially may not be relocated subsequently, making chemoprevention studies difficult. By using a special gastroscope, systematic mapping (4-quadrant biopsy specimens at 1-cm intervals) was performed in 22 patients (33 total mappings yielding 700 biopsy specimens). H&E, immunohistochemistry, and DNA ploidy analysis were performed. c-erbB-2 and positive Ki-67 were detected only in dysplastic sites; thus, their detection did not precede morphologically identifiable dysplasia. On the other hand, aneuploidy and p53 were detected in dysplastic and nondysplastic areas. p53 was correlated with dysplasia, and S-phase narrowly missed correlation, while aneuploidy was not correlated. PCNA and bcl-2 were ubiquitous, limiting their usefulness. On second maps, epithelial type was reidentified with 81% accuracy. A significant correlation was found between p53 and dysplasia. Sites of dysplasia and abnormal biomarkers could be relocated accurately by using endoscopic mapping. Therefore, mapping combined with biomarker studies may provide better surveillance and serve as a useful technique in chemoprevention studies. PMID- 11026102 TI - Mucoepidermoid carcinoma involving Warthin tumor. A report of five cases and review of the literature. AB - We describe 5 cases of mucoepidermoid carcinoma (MEC) involving Warthin tumor (WT) of the parotid gland. The WT size ranged from 1.7 to 6.0 cm. The MECs were much smaller, 0.3 to 1.7 cm. In 3 cases, the WT completely surrounded the MEC, and in 2 cases neither WT nor MEC surrounded the other. Each MEC was low grade, 3 grade I and 2 grade II. One MEC had evidence of vascular invasion. All patients underwent partial or subtotal parotidectomy with negative resection margins. Clinical follow-up (range, 8-52 months) for 3 patients showed no evidence of recurrence. The pathogenetic relationship between WT and MEC in these cases is uncertain. In 4 cases, foci of squamous or mucous metaplasia were found in the WT component, associated with mild cytologic atypia in 3 tumors. However, a direct transition from WT to MEC was not identified. In 1 case, MEC was present 45 months before WT, suggesting that the recurrent MEC involved WT coincidentally. The small size and low grade of the MEC and the negative resection margins most likely explain the good outcome for the 3 patients with clinical follow-up data available. PMID- 11026103 TI - Impact of qualified (indeterminate) diagnoses on the accuracy of renal, thyroid, and breast fine-needle aspiration biopsy. AB - A qualified (indeterminate) diagnosis (QD), such as "suggestive of malignancy," is thought to complicate patient management by heightening clinical uncertainty. We report that QDs increase the overall effectiveness of renal, thyroid, and breast fine-needle aspiration (FNA) biopsy and that the probability that a qualified diagnosis is negative (QDN) can be predicted by the formula QDN = number of QDs x (proportion of false-negative outcomes/disease prevalence expressed as a proportion). Results of renal (n = 24), thyroid (n = 163), and breast (n = 456) FNA biopsies performed from January 1992 through December 1998 were reviewed and correlated with results of tissue biopsies. For each body site, the FNA biopsies were placed into 1 of 2 diagnostic categories: unqualified diagnoses (UQDs) or QDs. Comparison of test performance characteristics for UQDs only and UQDs combined with QDs demonstrated that inclusion of UQDs increased FNA sensitivity and reduced FNA false-negative diagnoses. More important, the probability that a QD was negative could be predicted from test performance characteristics derived from UQDs. PMID- 11026104 TI - Hyaline globules in renal cell carcinoma and hepatocellular carcinoma. A clue or a diagnostic pitfall on fine-needle aspiration? AB - The cytologic similarity to hepatocellular carcinoma (HCC) of 2 cases of granular renal cell carcinoma (RCC) with hyaline globules (HGs) prompted us to evaluate the frequency of HGs in RCC and HCC and the association between cell type, differentiation, and malignant potential of these 2 neoplasms and the presence of HGs. We studied fine-needle aspirates from 23 cases of RCC (primary, 7; metastatic, 16) and 23 cases of primary HCC and noted anatomic site, tumor cell type, and presence, quality, and quantity of HGs. Fuhrman nuclear grade was assigned to RCC and overall differentiation to HCC cases. RCC cell type was granular (7), mixed (4), and clear (12). HCC cases were granular (22) and clear (1). Morphologically similar, predominantly intracytoplasmic HGs were identified in 4 RCCs and 10 HCCs. All 4 RCCs with HGs were at metastatic sites, granular cell type, and high Fuhrman grade. HGs showed no association with differentiation of HCC. While HGs are seen more commonly in HCC than in RCC, their presence in an adenocarcinoma should bring renal origin into consideration. The presence of HGs in metastatic granular RCC, particularly in the liver, can lead to misinterpretation as primary HCC. PMID- 11026105 TI - Nuclear accumulation of beta-catenin is a common and early event during neoplastic progression of Barrett esophagus. AB - Our aim was to characterize expression and mutation of beta-catenin in the progression of Barrett esophagus to adenocarcinoma. Immunohistochemical analysis of beta-catenin was performed on paraffin-embedded tissue from 30 cases with adenocarcinomas and premalignant lesions. To determine whether there is a correlation between beta-catenin nuclear accumulation and exon 3 mutation of this gene, mutational analysis by polymerase chain reaction-single-strand conformation polymorphism was performed on DNA extracted from the same 30 adenocarcinomas. As a result, the prevalence of reduced expression of beta-catenin on the membrane, with or without nuclear staining, increased significantly from low-grade (LG) to high-grade (HG) dysplasia. Focal nuclear staining for beta-catenin was present in 19 cases of adenocarcinoma, and nuclear staining was associated significantly with progression from metaplasia to LG dysplasia. In addition, in glands with clear histologic transition from metaplasia to LG dysplasia, nuclear accumulation of beta-catenin was found only in the LG dysplastic areas. No mutation in exon 3 of the beta-catenin gene was detected in adenocarcinomas. These results demonstrate that disturbance of the APC/beta-catenin pathway, as indicated by nuclear accumulation of beta-catenin, is a common and early event during neoplastic progression in Barrett esophagus. PMID- 11026107 TI - Cell block cytology. Improved preparation and its efficacy in diagnostic cytology. AB - Cell blocks prepared from residual tissue fluids and fine-needle aspirations can be useful adjuncts to smears for establishing a more definitive cytopathologic diagnosis. They can be particularly useful for categorization of tumors that otherwise may not be possible from smears themselves. A modified cell block technique using an improvised ethanol formalin fixative (Nathan alcohol formalin substitute) followed by a simple paraffin processing schedule is described. This improved preparation offers excellent cytomorphologic features corresponding closely to cells in Papanicolaou-stained smears and ensures optimal preservation of histochemical and immunocytochemical properties. The technique is simple and reproducible and uses routine safe laboratory chemicals. The efficacy of cell blocks also is discussed. PMID- 11026106 TI - Minocycline as a cause of drug-induced autoimmune hepatitis. Report of four cases and comparison with autoimmune hepatitis. AB - We describe the clinical and liver biopsy morphologic features for 4 patients with minocycline-induced autoimmune hepatitis (group 1). We compared the serum laboratory values and liver biopsy findings from group 1 with those from 10 patients with sporadic autoimmune hepatitis (group 2). All patients in group 1 had positive serum antinuclear antibody titers, but none had positive serum anti smooth muscle antibody titers. The morphologic findings of group 1 biopsies were those of autoimmune hepatitis in all 4 patients. In addition, 1 of these biopsy specimens also had scattered single eosinophils, unlike autoimmune hepatitis. The mean histologic activity index scores for patients in groups 1 and 2, respectively, were 6.7 and 5.4. No patients in group 1 had marked bridging fibrosis or cirrhosis, compared with 4 of 10 patients in group 2. Minocycline induced autoimmune hepatitis is usually identical to sporadic autoimmune hepatitis. The absence of eosinophils does not exclude the possibility of a minocycline cause. In the absence of clinical or morphologic differences, a recent ingestion of minocycline should be excluded before the diagnosis of sporadic autoimmune hepatitis is established. Whether the drug is unmasking latent autoimmune hepatitis is unclear. PMID- 11026108 TI - Detection of von Willebrand disorder and identification of qualitative von Willebrand factor defects. Direct comparison of commercial ELISA-based von Willebrand factor activity options. AB - Two von Willebrand factor (vWF):collagen binding (activity) assay (CBA) kit methods are commercially available. A monoclonal antibody (MAB)-based enzyme linked immunosorbent assay (ELISA) system reported to correlate with a standard vWF:ristocetin cofactor (RCof) assay is also commercially available. It is marketed as a vWF:Activity assay and is available in 2 assay version formats. In the present study, these 4 vWF-activity options were compared directly with in house vWF:CBA ELISAs for their ability to detect von Willebrand disease (vWD) and identify qualitative vWF defects. The 2 MAB-based systems detected vWD but could not specifically identify qualitative vWF defects, although the recently modified Mark II kit was more effective for the latter compared with the original Mark I kit. All vWF:CBA methods, including in-house and commercial, also effectively detected vWD but differed in their ability to identify qualitative vWF defects. Effectiveness was highest using the in-house reference vWF:CBA (using a type I/III collagen mix product from equine tendon), the Gradipore vWF:CBA (also uses equine tendon-derived collagen), or the in-house vWF:CBA methods using type III human collagen at a relatively low concentration (1 or 3 micrograms/mL, without covalent linkage). The IMMUNO vWF:CBA seemed to be the least effective among the vWF:CBA methods for detection of qualitative vWF defects. PMID- 11026109 TI - Elevated anti-annexin V antibody levels in antiphospholipid syndrome and their involvement in antiphospholipid antibody specificities. AB - To clarify the involvement of annexin V (ANX) in antiphospholipid antibody (APA) specificities, we studied antiANX antibodies (aANX) using 2 kinds of enzyme linked immunosorbent assay plates (plain and gamma-irradiated) and anti-beta 2 glycoprotein I antibodies (a-beta 2GPI) in 53 patients with antiphospholipid syndrome (APS). The incidence of aANX IgG-positive results in the autoimmune APS group was significantly higher than that of healthy control subjects. However, we could not demonstrate a significantly higher incidence in the infection- or drug induced group. Nor could we find an increased incidence of IgM isotype. When the 2 plates were compared, the discrepancies of positivity were demonstrated in both isotypes. We speculated that these discrepancies between the plate surfaces were attributed to the altered antigenicity of ANX. Although positivity of a-beta 2GPI was associated significantly with clinical manifestations, no significant associations were demonstrated between the incidence of aANX-positive results and clinical manifestations. We inferred that the involvement of aANX in the pathogenic mechanism of APS is unlikely. PMID- 11026110 TI - More on viral hepatitis. PMID- 11026111 TI - Changing with the times. PMID- 11026112 TI - America: love it and leave it. PMID- 11026113 TI - Job satisfaction and professional development of health information administration faculty. AB - To provide seamless care for patients while the health care industry undergoes changes, an integrated health information system must be developed. This effort affects the roles and responsibilities of practitioners and educators in health information administration (HIA). The changes may affect job satisfaction of HIA educators as they prepare entry-level practitioners for emerging roles. A national survey of HIA faculty's job satisfaction used the JDI/JIG scales and a researcher-developed instrument. The survey asked what job, institutional, and individual factors were significantly related to overall job satisfaction, whether there is a relationship between organizational fit and overall job satisfaction, and whether HIA faculty were pursuing terminal degrees, and what incentives and impediments they found to do so. Response rate was 78%. These faculty were very satisfied with their jobs. Work, pay, supervision, and co workers were significantly related to overall job satisfaction, with work and supervision accounting for 47% of the variance. Doctorally prepared faculty valued research more than did others, and tended to be in institutions perceived to value research. Although 79% reported that their institutions encouraged them to pursue advanced degrees, only 23.6% were pursuing doctorates. These findings may be useful in recruiting HIA practitioners into academia as well as for accreditation purposes. PMID- 11026114 TI - Faculty perspectives of health promotion in allied health curricula: results of a national survey. AB - More knowledge and skills related to health promotion/disease prevention are being required of health professionals. Allied health graduates entering the workforce will need academic preparation in these areas. This study assessed faculty perspectives on the status of health promotion content in allied health programs by a survey of 524 program directors. A questionnaire gathered information about perception of importance, level of representation, and method of delivery of health promotion content. Response rate was 41%. The majority of participants thought this content area was important for their programs and was highly or moderately represented in their curricula. Level of representation was significantly associated with requirement for accreditation. Programs in the Northeast were more likely to think health promotion was important and those in the West were more likely to have it integrated into their curricula. The most common delivery modality was guest lecture. Allied health is adopting health promotion curricular content, but further study is needed to assess programs and faculty expertise and identify strategies to ensure greater consistency of delivering material. PMID- 11026115 TI - The influence of learning styles on collaborative performances of allied health students in a clinical exercise. AB - With the increasing emphasis on interdisciplinary practice, there is a need to recognize factors that promote effective interdisciplinary teams. The influence of individual learning-style preference on collaborative performance was examined in 78 occupational and physical therapy students in a neuroscience course. They were randomly assigned in pairs to one of three subsets based upon their Kolb Learning Style Inventory scores (active experimenters vs reflective observers). The students viewed a videotape of a quadriplegic patient's physical examination and completed a collaborative exercise that required performance skills on all levels of Bloom's taxonomy of learning. Analysis of variance revealed no significant difference among learning-style subset pairs in performance (F2, 36 = 0.43, p > 0.05), but the exercise's overall grade correlated with the average of the pair's highest individual scores on two prior written exams (r = 0.631, p < 0.05), suggesting that background knowledge may predict performance better than learning styles do. It is suggested that differences in information-processing styles as measured by the Kolb inventory do not affect interdisciplinary team performance. PMID- 11026116 TI - Determinants of clinical performance in a physical therapy program. AB - In admission decisions, allied health programs evaluate various factors that may include overall grade-point average (GPA), science GPA, GPA in the most recent 60 hours of course work, volunteer/work experience, and performance during an admission interview. Most use mathematical formulas based on these parameters, but the rationale for weighting each factor is not always clear. To determine whether pre-admission variables can differentiate between good and poor clinical performances, the authors compared pre-admission data from 118 graduates of a physical therapy (PT) baccalaureate program with their clinical performances. GPA in the PT curriculum was included to determine whether it was related to clinical performance, which was taken as the average of overall performance scores assigned to each student by his or her instructors at four clinical education sites. The students were divided into quartiles by clinical scores. One-way ANOVA was used to identify differences between quartiles for each variable (alpha level of p < 0.05). The interview score was the only pre-admission variable that differentiated between students who did and did not perform well in the clinic (p < 0.003). Those who performed better in the clinic also had higher PT GPAs (p < 0.0001). The interview format and scoring system used may easily be adapted for other programs. Recommendations are made regarding how to utilize interview scores in the admission decision. PMID- 11026117 TI - Comparison of two teaching methods in a chiropractic clinical science course. AB - This study was done to determine whether the test performances of chiropractic students taught by traditional didactic methods (n = 50) and those undergoing problem-based learning (PBL) (n = 54) would differ. A 25-question multiple-choice test of knowledge synthesis was given to both groups of students. Although the novelty of the PBL approach was a cause of concern among the PBL students, who tended to be uncertain about what was expected of them and to lack confidence in the depth of their knowledge, their performance on the test was equivalent to that of the traditional students. The usefulness of PBL for educating allied health professionals adept at problem solving for future practice needs to be examined further. PMID- 11026118 TI - Equivalency of computer-based and paper-and-pencil testing. AB - This study examined the equivalence of computer- and paper-based versions of an examination through score differences across the two test formats as well as students' attitudes toward and perceptions of computer-based examinations. Thirty senior dental hygiene students were randomly divided into two equal groups. One took the first examination on computer, while the second took it on paper. Later, the groups were switched for a second examination. In completing the computer version, each student was asked to complete a survey that examined his or her experience with as well as attitude and perceptions toward computer-based testing. Students using the computer performed as well as or better than those using paper; the increase in performance was significant for the first examination (p < 0.05). Student acceptance of the computer format was mixed, possibly varying with prior exposures to such formats. Benefits of the computer based examination included reduction in time required for scoring and recording of grades, quicker student progression through the examinations afforded by the digitizing of the visuals, and ease of item analysis for both individual students and the group. PMID- 11026119 TI - Facilitating factors for, barriers to, and outcomes of interdisciplinary education projects in the health sciences. AB - Appropriate collaboration and communication among health professionals is viewed as increasingly important in meeting the challenges of today's health care system. Student socialization to interdisciplinary practice is thought to be facilitated through interdisciplinary academic and clinical learning experiences during the educational program. The purpose of this article is to identify factors commonly cited as facilitating and inhibiting interdisciplinary education, to provide insight for the development of successful interdisciplinary education projects. PMID- 11026120 TI - Pioneering Allied Health Clinical Education Reform: report of a National Consensus Conference. PMID- 11026121 TI - A path analysis of cognitive, affective, and behavioral predictors of treatment response in a methadone maintenance program. AB - PURPOSE: Continued illicit drug use by opioid-dependent patients maintained on methadone is a serious problem, undermining the goal of methadone maintenance treatment and increasing the risk for HIV. The current study employed structural equation modeling (SEM) to examine cognitive, affective, and behavioral predictors of treatment response during the first 12 weeks of methadone maintenance treatment. METHODS: 302 opioid-dependent individuals (72% male; 28% female) entering a methadone maintenance program (MMP) were provided with a comprehensive intake assessment. Heroin and cocaine use were assessed using twice weekly urine toxicology screens. The model accounted for 37 percent of the variance in abstinence from illicit opiates and 38 percent of the variance in abstinence from cocaine. RESULTS: Continued heroin use while maintained on methadone was predicted by pre-treatment severity of addiction and by strength of self-identity as an "addict." Cocaine use was predicted by pre-treatment severity of addiction, low self-efficacy, lack of negative affect, and strength of self schema. The strongest predictor of abstinence from both heroin and cocaine was attendance at the adjunctive manual-guided psychosocial group interventions. IMPLICATIONS: Implications for treatment are discussed. PMID- 11026122 TI - Smoking behavior in a smoking workplace. AB - PURPOSE: This report addresses whether smoking increases as a function of working in a smoking environment. METHODS: Casino employees were targeted to complete a survey designed to assess smoking behavior. The survey contained standardized questions consistent with other major surveys on tobacco use to ensure comparability with state and national trends. Data collection occurred between September and November 1997. RESULTS: Out of 755 surveys distributed, 587 completed surveys were returned, resulting in a response rate of 77 percent. A test of proportions showed that the proportion of smokers had not changed significantly as a result of working in a smoking environment although the amount of smoking decreased significantly (one-sample t-test). The change in the amount of smoking was assessed using difference scores. Males and females did not differ in the rate in which their smoking changed. A significant correlation between age and the change score suggested that younger smokers were more likely to increase their intake whereas the older smoking employees were more likely to decrease their consumption by the time of the survey. IMPLICATIONS: The finding that individuals who work in a smoking environment decreased their smoking behavior merits further investigation in the area of smoking aversion. PMID- 11026124 TI - Couples' drinking patterns, intimate partner violence, and alcohol-related partnership problems. AB - PURPOSE: Past research has revealed a strong correspondence between alcohol use and family dysfunction (e.g., Straus and Gelles in 1986 and 1990). Unfortunately, much of this research has relied on reports from only one family member. METHODS: Here, we present the results from a recent, nationwide study of alcohol consumption patterns, alcohol-related partnership problems, and intimate partner violence (IPV) based on reports from both romantic partners of 1,615 married and cohabiting couples. RESULTS: Using the "drinking partnerships" construct developed by Roberts and Leonard in 1997, we found that despite considerable concordance between couple members' drinking behaviors, discrepant drinking patterns were strongly predictive of relational distress and the incidence of physical violence. IMPLICATIONS: These findings suggest that the interaction between couple members' alcohol-related behaviors has crucial implication for the health and well-being of the entire family. PMID- 11026123 TI - Perception of traffic accident risk and decision to drive under light alcohol consumption--a double-blind experimental study. AB - PURPOSE: Public traffic safety campaigns in Germany have focussed on the changing risk perception of young drivers. While there is some consensus that perceptions of risk affect driving, less is understood about the relationship and interaction of alcohol consumption and risk perception on the decision to drive. We examined the influence of light alcohol consumption on risk perception and decision to drive, and the interaction of alcohol consumption and cognitive feedback on the handicapping effect of alcohol on risk perception and decision to drive. METHOD: In a double-blind block-randomized experimental study of 104 young drivers between 19 and 24 years of age, with two experimentally manipulated independent factors of alcohol consumption (three levels: 0% BAC, 0.015% BAC, 0.03% BAC) and feedback (positive or negative), we assessed three dependent variables: perception of traffic accident risk, subjective judgement about driving-relevant cognitive performance, decision to drive a car. Analyses of variance and covariance were used to analyze differences between levels of experimental factors. RESULTS: We found that persons with 0.015 BAC performed better than persons in both other alcohol conditions on the standardized risk perception task. Perceived handicap of driving was significantly more pronounced for negative feedback compared to positive feedback with no influence of the level of alcohol consumption. No significant influence on decision to drive was found of either level of alcohol consumption, feedback or sex. IMPLICATIONS: Decision to drive in young drivers could not be influenced by feedback or light consumption. Public health approaches have to find better determining factors. PMID- 11026125 TI - Service use and cost by mentally ill chemical abusers: differences by retention in a therapeutic community. AB - PURPOSE: Earlier research estimated the incremental costs and outcomes of a modified therapeutic community (modified TC) for mentally ill chemical abusers (MICAs) relative to a treatment-as-usual (TAU) control group. The present study extended the cost analysis by disaggregating the modified TC group into clients who completed the program (completers) and clients who dropped out (separaters). METHODS: Bivariate and multivariate analyses were conducted to estimate differences in treatment and other service costs among completers, separaters, and TAU. Subjects were sequentially assigned to the modified TC (n = 171) or TAU (n = 47), and the analysis period covered 12 months post-baseline. Using a standardized instrument to collect resource use and cost data, the estimated weekly cost per client in the modified TC was $554, with completers showing a larger average cost of treatment ($27,595) than separaters ($9,986). RESULTS: The average TAU subject had a much higher cost for other (non-modified TC) services ($29,795) relative to separaters ($22,048) or completers ($1,986). These findings suggest that, from baseline to the 12-month follow-up, the total cost of modified TC treatment and other services for completers may be slightly lower than the total cost for separaters or TAU subjects. Since the modified TC group had better outcomes than the TAU group, and the completers had better outcomes than the separaters, the modified TC program could be an effective mechanism to reduce the costs of service utilization as well as improve clinical outcomes. IMPLICATIONS: This detailed investigation into service utilization and cost provides policy makers and program directors with valuable information regarding potentially cost effective interventions and further underscores the importance of retention in treatment for this vulnerable population. PMID- 11026126 TI - Short form of a situational temptation scale for heavy, episodic drinking. AB - PURPOSE: A short form for situational temptations to drink scale was developed from an original 21-item inventory by Migneault. METHODS: The form measured four hypothesized subscales of temptations on a sample of 348 college drinkers (66% female). Peer pressure, social anxiety, negative affect, and positive/social situations subscales were replicated and reduced. RESULTS: Strong empirical support was found for a hierarchical model, indicating that the four subscales can be summed to provide a global measure of situational temptations. Confirmatory factor results, internal and external validity, and high correlations with the original measures indicate that the short form was as psychometrically valid as the original measure. IMPLICATIONS: Measures of external validity demonstrated the applicability of this measure to heavy drinking prevention programs. PMID- 11026127 TI - Sexual identity and alcohol-related outcomes: contributions of workplace harassment. AB - PURPOSE: While workplace sexual harassment has received a great deal of attention in both the popular media and scientific literature, less attention has been directed to the differential occurrence of sexual harassment among lesbians, gay men, and heterosexual men and women, and the relationships between these experiences and alcohol-related outcomes. Additionally, the distribution of alcohol-related outcomes of non-sexual forms of workplace harassment among these groups have not been adequately explored. METHOD: Using data from a university based study of workplace harassment and alcohol use (N = 2492), we focus on exposure to workplace harassment and alcohol-related outcomes for lesbians, gay men, and bisexuals compared to heterosexual men and women. RESULTS: Lesbian/bisexual women did not differ significantly from heterosexual women in their experiences of workplace harassment. However, stronger linkages between harassment and increased alcohol consumption and problems were found for lesbian and bisexual women than for heterosexual women. Gay/bisexual men, on the other hand, experienced significantly more sexual harassment than heterosexual men, but did not report a corresponding increase in alcohol use and abuse. IMPLICATIONS: Implications for future research on sexual identity, alcohol use, and workplace harassment are discussed. PMID- 11026128 TI - Psychometric evaluation of the short form inventory of drinking situations (IDS 42) in a community-recruited sample of substance-abusing women. AB - PURPOSE: We investigated the psychometric properties (factor structure, internal consistency reliability, concurrent validity) of the Short Form Inventory of Drinking Situations (IDS-42) in women substance abusers. METHODS: A sample of 297 substance-abusing women was recruited from the community. The women completed the IDS-42 and the three-factor Drinking Motives Questionnaire (DMQ). RESULTS: Confirmatory factor analyses of IDS-42 items suggested a hierarchical structure for the scale. Eight factors (corresponding to Marlatt and Gordon's eight heavy drinking situations) provided the best model fit at the lower-order level, and three factors (i.e., Negatively Reinforcing vs. Positively Reinforcing vs. Temptation Situations) provided the best model fit at the higher-order level. Lower- and higher-order IDS-42 subscales were shown to possess adequate-to-high levels of internal consistency. The eight lower-order IDS-42 factors demonstrated excellent concurrent validity with conceptually similar DMQ subscale scores. Across the entire sample of female substance abusers, a higher frequency of heavy drinking was reported in Positively Reinforcing Situations and Unpleasant Emotions Situations, as compared to other heavy drinking situations. IMPLICATIONS: Results support the IDS-42's good psychometric properties and demonstrate its utility as a tool in identifying situation-specific antecedents to heavy drinking among women substance abusers. PMID- 11026129 TI - [Epilepsy as a clinical and neurophysiological problem]. AB - On the basis of both own investigations and publications the next basic aspects of epilepsy were discussed: a) a clinical one--definition of epilepsy, seizures' classification, children's and women's epilepsy, status epilepticus; b) a neurophysiological aspect--epileptic and antiepileptic mechanisms, the significance of slow EEG activity, interhemispheric interactions, influence of the seizures on brain homeostasis. PMID- 11026130 TI - [Cerebrogenic disorders of cardiac rhythm and conductivity in epilepsy patients]. AB - 51 epileptic patients aged 16-32 years without any signs of cardiovascular pathology were examined. The primary generalized epilepsy was diagnosed in 24 cases while a partial one--in 27 patients. Were examined a state of the heart (echo- and electrocardiography, Holter's monitoring of ECG), of the brain (computed echoencephalography, visual evoked potentials) and of the autonomic nervous system (variability of cardiac rhythm, autonomic tonus). A control group included 15 healthy individuals. Significantly higher prevalence of the disorders of the cardiac rhythm and conduction was found in patients with epilepsy as compared with a control group (p < 0.001). More frequently there were sino-atrial blocks of the II degree of the I type (p < 0.01), heterotopic rhythms (p < 0.05), supraventricular extrasystole (p < 0.1), supraventricular paroxysmal tachycardia (p < 0.05), supraventricular migration of the pacemaker (p < 0.01). It was found that close relationship between cardiac arrhythmias and epilepsy was mobided by a complex of pathophysiologic mechanisms. Among them a leading role belongs to dysfunction of suprasegmental autonomic centers with a predominant activation of sympathetic or parasympathetic departments of the autonomic nervous system. Significant arrhythmogenic factors appeared to be: presence of organic damages in the brain, frequency of epileptic fits, dysfunction of suprasegmental autonomic centers with activation of cerebral ergotropic systems. PMID- 11026131 TI - [The prevalence of cardiovascular pathology in alcoholism patients]. AB - ECG was made in 336 patients with alcoholism that were admitted in a special hospital for alcoholic patients during one year. 146 patients were examined in the state of alcohol withdrawal syndrome, after its arrest and in stress. A prevalence of ECG changes well as stability of such alterations, prognostic criteria for a further course of both alcoholism and cardiovascular pathology were determined. A correlation was established between the cardiovascular changes on the one hand the age of the patients and the type of a course of alcoholism on the other hand. PMID- 11026132 TI - [Pathological reactions of the erotic possession type]. AB - The paper presents an analysis of the disorders with manifestations of non delirious pathologic love as a response to the action of psychogenic factor on the basis of constitutional predisposition. By the results of the examination of 31 patients with prolonged psychogenic depressions, characterized by the phenomenon of erotic possession, 2 types of the pathologic reactions were distinguished: with a predominance of the possessed images and with a predominance of the delirious-like fantasies. Psycho-traumatic situations that reflected loss of significant relations (either real or imaginary) as a result of the divorce, broken relations, "love triangle", etc. were investigated. Estimation of the premorbid state in the patients allowed both to reveal accentuation in a sphere of love relationship and to single out a main mechanism of the development of pathologic reactions (by type of "key" experience) as well as to define typological differentiation of psychogenic by type of erotic possession taking into consideration the constitutional anomalies. PMID- 11026133 TI - [The optimization of epilepsy therapy with finlepsin]. AB - A clinical pharmacological study (270 patients) was made to optimize therapy of epilepsy with carbamazepines (finlepsin and its retard form). It was established that insufficient (standard) doses of finlepsin are often used, serum levels of the drug are not estimated, drug pharmacokinetics is not taken into account, maximum permissible doses of the drug are not reached. Estimation of the efficiency of both preparations in the treatment of various types of the seizures was carried out. The advantages of finlepsin-retard were demonstrated. PMID- 11026135 TI - [The humoral immunity characteristics of women with neurotic depressions]. AB - The levels of autoantibodies and their anti-idiotypes to proteins of nervous tissue (S100b, GFAP, MP65 and nerve growth factor) were studied in 85 women of 18 48 years of age with psychogenic neurotic depressions. It was found that the changes of the levels of autoantibodies to the proteins of nervous tissue correspond to various somatoautonomic manifestations and correlate with severity of depression, its typological variations and duration of the disease. The data obtained show a close relationship between nervous and immune systems that was expressed by some parallelism of the dynamics of the changes of both neurotic depressive symptoms and serum immunoreactivity to neuroantigens. PMID- 11026134 TI - [Akatinol memantin in Alzheimer's disease: clinico-immunological correlates]. AB - 16 patients with Alzheimer's disease were treated with akatinol memantine. The levels of production of interleukin-1 (IL-1) in lymphocyte culture were evaluated both before and after the treatment that lasted for 2-4.5 months. Before the therapy a significant (p < 0.05) increase in the production of IL-1 was observed in patients with mild and moderate dementia, with early onset of the disease and with the duration of the disease up to 9 years as compared with a control group (24 healthy individuals). After the treatment the production of IL-1 was decreased in patients with moderate dementia, with early onset, and with hereditary form of the disease. Decrease of the level of IL-1 production after the treatment corresponded to improvement of the cognitive functions of the patient. In cases of unsatisfactory treatment (without any improvement of a clinical state) IL-1 production was at higher levels (p < 0.05) as compared with the control values. It is suggested that initial high level of IL-1 production could serve as a predictor of efficiency of therapy with akatinol memantine in patients with early onset of Alzheimer's disease. PMID- 11026136 TI - [The clinico-epidemiological characteristics of epilepsy in the Jewish Autonomous Province]. AB - In Jewish Autonomous Province an investigation of epilepsy was performed, including all the patients that lived on this territory. 547 epileptic patients were defected, that was 2.9 cases per 1000 of the population. Estimation of the patients was performed using 65 standard signs. The data were received concerning prevalence of epilepsy in the region, sex-age structure of the patients, clinical manifestations of the disease and other parameters. Special attention was paid to clinical-genetic aspects of epilepsy. The data received are important for carrying out a clinical examination and for differential rehabilitation of the patients, for planning medical prophylaxis measures as well as for medical genetic consultative service. PMID- 11026137 TI - [Temporal lobe epilepsy]. PMID- 11026138 TI - [A clinico-immunological study of diphtheritic polyneuropathy]. PMID- 11026139 TI - [Biological testing for dysfunctins extracted by cerebrospinal fluid sorption in heroin addiction]. PMID- 11026140 TI - [The use of Seroquel in schizophrenia]. PMID- 11026141 TI - [Post-stroke epilepsy]. PMID- 11026142 TI - [HCV genotypes. Liver histopathology and immunologic profile in four groups of patients]. AB - AIM: Study on the prevalence of MCV genotypes, and correlation with liver pathology and immunological parameters. PATIENTS AND METHODS: 77 chronic hepatitis C patients (52 males, 25 females), mean age 44 +/- 14 years, belonging to four groups: (1) 23 (11 males) without other aetiology, (II) 19 (18 males) excessive drinkers, (III) 18 (12 males) haemodialysed, (IV) 17 (11 males) renal transplantation patients Genotyping was done by PCR (primers of the core, and hybridisation with specific probes). Serum Igs A, G, and M (by nephelometry), and peripheral blood lymphocyte (PBL) subsets (by flow cytometry) were determined. RESULTS: One genotype was found in 62 patients (1b 69.2%, 1a 21.5%, 2a 3.1%, 2b 3.1%, 3a 9.2%), and two genotypes in 4 patients (1b + 3a in 1 of group 1, 1b + 2a in 2 and 1a + 3a in 1 of group 111). Twelve cases (15.6%) were not identified (NI). Relative prevalence was not different in the four groups, but in 7 drug addicts 1 b was not found (Ia in 71.4%, 3 a 28.6%) The relationship between genotypes and age was significant (p < 0.05): in the 34 patients with less than 40 years, 1b was found in 38.2%, in the others in 41.2%, and NI in 20.6%; in those with 40-60 years, 1b was found in 68.8%, in the others in 15.6%, M in 15.6%; in those above 60 years, 1b was found in 90.9%, in the others in 9.1%. Of those identified, only genotype 1 (1a and 1b) was associated to moderate or severe activity, and infected 11/13 cases of cirrhosis or severe fibrosis. IgM (g/dL) was lower in 1b than in the others: 1.58 + 1.23 vs. 2.53 + 1.93 (p < 0.01). PBL (per mm3) were lesser in 1b than in the others, with significance for the CD8+: 540t239 vs. 739 + 420 (p < 0.01). CONCLUSIONS: Genotype 1b was the most prevalent in Portuguese patients, more significantly in the elderly, and was absent in drug addicts. The prevalence of genotypes is similar in general patients, in chronic haemodialysed, in renal transplantation recipients and in alcoholics. More severe liver pathology was associated with 1b and 1a genotypes. IgM and CD8+ had lower mean values in 1b infected patients. Other genotypes are certainly important in Portugal. PMID- 11026144 TI - [Hospital discharge letter --Clinical information for the family physician]. AB - INTRODUCTION: There are aspects related with the proportion and quality of discharge letters which may influence the management of shared health care after hospital discharge; we describe them taking into consideration that no changes regarding the usual way of giving hospital discharge took place. AIMS: To determine the proportion of cases in which the hospital discharge of inpatients was accompanied with a discharge letter and to measure its quality. METHODS: A descriptive and transverse observation study referring to the year of 1996. The authors observed, in the clinical files of this Department, the total cases of discharge of inpatients with ischaemic heart disease (123 cases). Besides measuring the variable "discharge letter existence", we studied the composed variable discharge letter quality (existence of clinical history data, objective observation data, complementary test results, diagnosis, therapeutic procedures, and also the readability of the clinical information and the existence of the legible name of the specialist). RESULTS: From the study of 121 cases of discharge of inpatients with ischaemic heart disease, 90.2% presented discharge letters. When studied, their quality was considered sufficient in 57.7% and insufficient in the rest of the cases. We point out that 29.7% of the discharge letters did not present objective observation data. DISCUSSION: The clinical information provided in this Department, at the time of hospital discharge, presents a higher proportion and quality than that observed in other studies. PMID- 11026143 TI - [Evaluation of microscopy methods for the diagnosis of bacterial vaginosis]. AB - The diagnosis of Bacterial Vaginosis has always been controversial. During many years, the laboratory diagnosis of this syndrome was based on the criteria of Amsel et al (1983). This includes many factors, such us aqueous vaginal discharge, positive KOH test and the presence of clue-cells in a wet mount or Gram stain. Lately, a new diagnostic method only based on laboratory findings was performed by Nugent et al (1991), which has the advantage of being more objective and rapid. It is also easy to be used in any laboratory or even in a doctor's room. In this study, we evaluated 74 Gram stained vaginal smears and compared both Amsel et al (1983) and Nugent et al (1991) methods. Bacterial Vaginosis was diagnosed in 28% by Amsel et al (1983) criteria and in 31% by the Nugent et al (1991) criteria. The latter method seems to have a higher efficacy in diagnosing Bacterial Vaginosis, although both techniques together diagnose a higher number of cases. PMID- 11026145 TI - [Changes in cognitive function performance in patients after anesthesia]. AB - This study evaluates the mental performance of 112 patients (ASA grade 1 and 2) submitted to Orthopaedic, Urologic, Gynecologic, General and ENT surgery in Santo Antonio General Hospital. These patients received different types of anaesthesia: General Balanced Anaesthesia--GBA (n = 80), Total Intravenous Anaesthesia--TIVA (n = 17) and Regional Anaesthesia--RA (n = 15). Cognitive function was tested using the mini mental state test (MMS), which was done 24 hours before and 24 hours after anaesthesia. There was a high correlation between pre and postoperative MMS scores (R = 0.94). Patients in the GBA group show a significant (p < 0.001) decrease in cognitive performance 24 hours after surgery in comparison to the preoperative test. No differences were observed between the pre and postoperative MMS tests in the TIVA and RA groups. PMID- 11026146 TI - [Cluster headache]. AB - We present a review of 60 cases of cluster headache. Most patients were males, ranging from 19 to 65 years of age at the time of the first visit. Headaches consisted of short-lasting (from 15 to 210 minutes), intense, unilateral pain attacks, most frequently in the periorbital area, with ipsilateral autonomic signs (rhinorrhea, ptosis, tearing and conjunctival injection). Between attacks, patients were completely free of pain. The attacks occurred in bouts lasting 1 to 6 months, in which patients had daily headaches (one to three times a day). Headaches responded well to oxygen or ergotamine. Prophylactic therapy in most cases consisted of verapamil, also with a good response. We present this review in order to draw attention to this relatively rare form of headache with a specific therapy. PMID- 11026147 TI - [Atherosclerosis. Old problem, new perspectives]. AB - Atherosclerosis is the major cause of mortality in the population of the, so called, developed countries of western culture. Since the first half of this century, hypercholesterolemia was the hallmark for the investigation of atherosclerosis, improving the level of knowledge about the complex metabolism of lipoproteins. The occurrence of atherosclerosis in normolipidaemic subjects, and the relationship between this illness, other dysmetabolic features and certain infectious agents, led to the reformulation, in the last decade, of the pathophysiological archtypes, atherosclerosis was included in the group of the inflammatory processes. The inflammatory response to aggression of the arterial wall is the innovative issue of atherosclerosis investigation and laboratory follow-up in the new millennium. PMID- 11026148 TI - [Chronic myeloid leukemia]. AB - The authors describe a clinical case of chronic myeloid leukemia with a typical presentation (massive splenomegaly and marked leucocytosis), but with a complex translocation--t (7; 9; 22) (q11; q34; q11). The evolution was rather atypical with a lymphoblastic crisis in association with osteolytic lesions and with parenchymal and soft tissue lymphoblastic tumours. PMID- 11026149 TI - [Acute psychotic disorder associated with vigabatrin]. AB - Vigabatrin, a drug mainly used in the treatment of therapy-resistant epilepsy, has been associated to a large range of adverse effects, including acute psychotic disorders. The authors report the case of a 44-year-old patient with drug-resistant temporal lobe epilepsy that developed an acute psychotic episode about three weeks after initiating medication with vigabatrin. After the withdrawal of vigabatrin and the addition of an antipsychotic drug, the clinical picture improved promptly and the patient was discharged within a short period of time. The authors discuss the patient's diagnosis, the possible mechanisms of vigabatrin-associated psychosis and the therapeutic approach, stressing the questions that concern vigabatrin prescription and withdrawal. PMID- 11026150 TI - [Thoracic endometriosis]. AB - Endometriosis was first described by Russel more than one-hundred years ago and still remains a clinical entity of difficult comprehension, with totally aberrant symptomatology, particularly in extra-genital situations. The present article describes a clinical case of recurring right-sided catamenial pneumothorax in a 45 year-old caucasian woman. In her last episode of right chest pain the X-ray film showed, besides the presence of a pneumothorax, a nodular image, later identified by CT scan as being a transdiaphragmatic hernia with hepatic content. The patient was then submitted to diagnostic and therapeutic thoracotomy and the diaphragmatic endometriosis was confirmed. Finally, a total hysterectomy with bilateral salpingo-ophorectomy was performed and the patient's condition remained uneventful with combined hormone therapy. The clinical presentation, pathogenesis, and therapeutic resolution of this nosological entity are briefly discussed. PMID- 11026151 TI - [Development of the Portuguese version of MOS SF-36. Part II --Validation tests]. AB - This paper describes the study aimed at validating the Portuguese version of the MOS SF-36 instrument of assessment. It starts by presenting the results of the implementation of this instrument in a sample of 930 pregnant women and the results of scaling tests, including the values of internal consistency and reliability. However, since a reliable instrument is not necessarily a valid one, the results of several validity tests are also presented. Finally, this paper ends by recommending the use of the Portuguese version of the SF-36 instrument of assessment. PMID- 11026152 TI - [Medical oath and the notion of human dignity]. AB - Considering the recent commemoration of the 50th anniversary of the Proclamation of Human Rights, the author attempts to establish a parallel between the notion of human dignity as we read in the Hippocratic Oath, and the prevailing concept within the societies that either preceded, were contemporary or succeeded it. With that goal in mind, three ancient civilizations are focussed--Egypt, Mesopotamia and Greece--showing the differences in behaviour towards the human being, far different from the perspective of the Oath. The new view of human dignity and its due respect, in any circumstance, was only completely understood with the appearance and acceptance of Christianity. The author then draws attention to what may be viewed as the main feature of this century, as it approaches its end: the urge to lead the international community to accept, in a clear way, what Hippocrates defined in Medicine--the dignity of the human person, regardless of race, religion, sex or social class. PMID- 11026153 TI - HIV risk and prevention in emergency-affected populations: a review. AB - While basic guidelines on HIV prevention in emergencies have been available for several years, international agencies involved in the provision of health services have not placed sufficient priority on the prevention of the human immune deficiency virus (HIV) and other sexually transmitted infections (STIs) in complex emergencies. This paper reviews the factors that may increase the risk of HIV transmission in populations affected by complex emergencies and outlines recommendations for research and programmes. Research into the most appropriate methods of carrying out HIV surveillance and interventions in these settings is needed. In the post-emergency phase programmes need to be far more extensive than those offered under the Minimal Initial Services Package (MISP). While the potential for stigmatization represents an important constraint, there is a need to prioritize HIV/STI interventions in order to prevent HIV transmission in emergency-affected populations themselves, as well as to contribute to regional control of the epidemic. PMID- 11026154 TI - Conflict-induced displacement and involuntary resettlement in Colombia: putting Cernea's IRLR model to the test. AB - This paper tests Cernea's (1997) impoverishment risks and livelihood reconstruction (IRLR) model in cases of conflict-induced displacement (CID). In applying the model to a situation involving internal conflict, the article illustrates the particular problems encountered by internally displaced people (IDPs) and policymakers charged to respond to them. The article searches for local interpretations of CID and resettlement through a comparative profile of two IDP settlements in Colombia: one urban, the other rural. It concludes that the IRLR model, when contextualised, provides a useful tool to identify and categorise risks of impoverishment and resettlement priorities. At the same time, however, the article demonstrates that the model insufficiently captures the root causes or causality of CID. PMID- 11026155 TI - The use of cluster sampling to determine aid needs in Grozny, Chechnya in 1995. AB - War broke out in Chechnya in November 1994 following a three-year economic blockade. It caused widespread destruction in the capital Grozny. In April 1995 Medical Relief International--or Merlin, a British medical non-governmental organisation (NGO)--began a programme to provide medical supplies, support health centres, control communicable disease and promote preventive health-care in Grozny. In July 1995 the agency undertook a city-wide needs assessment using a modification of the cluster sampling technique developed by the Expanded Programme on Immunisation. This showed that most people had enough drinking water, food and fuel but that provision of medical care was inadequate. The survey allowed Merlin to redirect resources earmarked for a clean water programme towards health education and improving primary health-care services. It also showed that rapid assessment by a statistically satisfactory method is both possible and useful in such a situation. PMID- 11026156 TI - Internal displacement in Burma. AB - The internal displacement of populations in Burma is not a new phenomenon. Displacement is caused by numerous factors. Not all of it is due to outright violence, but much is a consequence of misguided social and economic development initiatives. Efforts to consolidate the state by assimilating populations in government-controlled areas by military authorities on the one hand, while brokering cease-fires with non-state actors on the other, has uprooted civilian populations throughout the country. Very few areas in which internally displaced persons (IDPs) are found are not facing social turmoil within a climate of impunity. Humanitarian access to IDP populations remains extremely problematic. While relatively little information has been collected, assistance has been focused on targeting accessible groups. International concern within Burma has couched the problems of displacement within general development modalities, while international attention along its borders has sought to contain displacement. With the exception of several recent initiatives, few approaches have gone beyond assistance and engaged in the prevention or protection of the displaced. PMID- 11026157 TI - The urban poor in Dhaka City: their struggles and coping strategies during the floods of 1998. AB - Bangladesh experienced one of the worst floods in recorded history in 1998. This paper focuses on the needs and coping strategies of the urban poor in Dhaka City, which had been very badly affected. The city's roads were completely under water, and most areas were water-logged with drainage and sewage systems blocked. Rising water levels compelled many slum dwellers to move to temporary shelters and relief camps. Women and children were the worst affected. The lack of sanitation facilities and privacy forced women and children to defecate in their own homes. There was an acute scarcity of safe drinking-water, and food prices rose dramatically. Diarrhoea, fever and colds were the most common illnesses affecting the poor. The floods left many of them unemployed, and in some families, the result was increased tension and incidents of domestic violence. In some areas, members felt pressured to repay micro-credit loans. Most NGOs, however, suspended loan repayments. During this period, a committee was set up to co-ordinate and work towards addressing some of the main post-flood problems. PMID- 11026158 TI - Let them eat risk? Wealth, rights and disaster vulnerability. AB - Disaster-vulnerability reduction is an impure public good: when provided to one it is provided to others, but not equally provided to all. This means that in addition to the question of how much disaster-vulnerability reduction to provide, policymakers face the question of to whom it should be provided. This essay distinguishes between two broad classes of approaches to the latter question, one based on wealth, the other on rights. PMID- 11026159 TI - An earthquake disaster in Turkey: an overview of the experience of the Israeli Defence Forces Field Hospital in Adapazari. AB - On 17 August 1999 at 3:04 a.m., an earthquake of 7.4 magnitude (Richter scale) struck the Marmara region in Turkey. The city of Adapazari suffered 2,680 fatalities with approximately 5,300 injured. The Israeli Defence Forces (IDF) field hospital arrived at Adapazari, on day four after the quake. The team consisted of 102 personnel. The field hospital acted as a secondary referral centre. A total of 1,205 patients were treated in the field hospital between day four and day 14 of the earthquake. The frequency distribution of the medical problems seen in the field hospital was 32 per cent internal medicine, 13 per cent general surgery including plastic, 21 per cent orthopaedic surgery, 23 per cent paediatric disease, 10 per cent obstetrics and gynaecology and 1 per cent major psychiatric disorders. A mean number of 35 patients per day were hospitalised in the field hospital for between 24 hours to one week. The rapid establishment of the field hospital enabled the local medical facilities to 'buy time' in order to organise and restore surgical and hospitalisation abilities in this disastrous situation. PMID- 11026160 TI - Casualty treatment after earthquake disasters: development of a regional simulation model. AB - This paper presents a new approach to the casualty treatment problem following a large-scale disaster, based on a mathematical model of how a regional health-care system responds to an earthquake event. The numbers and locations of casualties rescued alive, the scale of pre-hospital care, the post-earthquake hospital capacity, and the transport system are inputs to the model. The model simulates the movement of casualties from the stricken areas to hospitals. It predicts the number of casualties that die as well as other statistics about the health-care system response, such as waiting time before treatment. The model can be run with varying input assumptions to simulate alternative disaster response strategies. Preliminary runs demonstrate the potential of the model as a tool for planning and training. PMID- 11026161 TI - General and specific measures of quality of life--do we need both? PMID- 11026162 TI - Measurement of staff empowerment within health service organizations. PMID- 11026163 TI - Using vignettes to study nurse practitioners' performance in suspected domestic violence situations. AB - Vignettes have often been used to evaluate students or collect data in nursing research. The format is familiar to most nursing students as well as nurses and nurse researchers. This article presents the development and testing of the Nurse Practitioner Performance Tool (NPPT) which used vignettes as an approach to nurse practitioner performance evaluation. In this example, vignettes were used in a quasi-experimental design to collect data from Adult and Family Nurse Practitioners (A/FNP). The focus was on the diagnosis and intervention performance of the A/FNPs when addressing suspected cases of domestic violence. PMID- 11026164 TI - Factor structure of the Reynolds Adolescent Depression Scale in a sample of school-based adolescents. AB - Depression is a common health problem in the adolescent population. The Reynolds Adolescent Depression Scale (RADS) is used to measure depression in clinical and community adolescent samples. Although there is available evidence for the reliability and validity of the RADS, there is insufficient documentation of its factor structure. This study examined the factor structure of the RADS in adolescent boys and girls (m-144). Internal consistency reliability ranged from .91 to .94 based on grade level, and was .91 for boys and .93 for girls. Factor analysis resulted in a 5-factor solution. Interpretation of factors were as follows: (a) Factor I--generalized demoralization; (b) Factor II--despondency and worry; (c) Factor III--externalized somatocism; (d) Factor IV--anhedonia; and, (e) Factor V--self-worth. PMID- 11026165 TI - Measuring continuity of elders' posthospital care. AB - Continuity of care is a critical component of quality patient care, yet the paucity of reliable and valid measures of continuity of care make it difficult to ascertain the extent to which continuity has been achieved. The purpose of this article is to describe the development of an instrument to measure continuity of care that incorporates the perspectives of elders hospitalized for a chronic illness and their family caregivers. The instrument was used, and its reliability and validity examined, in a series of studies related to elders' posthospital transition. Elders in the studies ranged in age from 55 to 94 years. The findings supported content and construct validity, internal consistency reliability, and ability to detect changes in the same subjects at different points in time for the care management and services subscales. With further refinement, the continuity of provider and conflicting information subscales might also facilitate assessment of care continuity. PMID- 11026166 TI - Assessing abuse among the elderly living in public housing. AB - Abuse and victimization among the elderly have received increased attention in the last few years from practitioners, researchers, and policymakers but remain growing problems, especially for elders who live in public housing. This study assessed the psychometric properties of an existing instrument, the Hwalek Sengstock Elder Abuse Screening Test (HSEAST), which has been tested in only two studies. The instrument was administered to 100 African American, Hispanic, and White elders living in public housing. Study results were compared to findings from a previous study of elderly groups. A principal components factor analysis of the 15-item instrument supported the 3-factor structure for a total of 10 items (factor loadings = 0.4 or >), explaining 38% of the variance. A discriminant function analysis showed that 6 items were as effective as the 9 item model in classifying cases (71.4%) as abused. PMID- 11026167 TI - A Norwegian version of the Incontinence Stress Questionnaire-Staff Reaction: translation and validation for cross-cultural use. AB - The aim of the present study was to translate and validate the Incontinence Stress Questionnaire--Staff Reaction (ISQ-SR) for cross-cultural adaptation to Norway. The ISQ-SR is a 30-item questionnaire developed in the U.S.A. by Dr. Lucy C. Yu. The questionnaire examines the experience of nursing home staff when working with patients with urinary incontinence. This article describes both the translation and testing procedure. Statistical analyses were carried out in the following steps of the testing procedure: (a) bivariate examination, (b) principal components factor analysis, (c) determination of internal consistency, (d) frequency analysis, and (e) estimation of test-retest reliability. Finally, the translated and tested questionnaire was compared with the original version. The Norwegian version consists of three factors with a total of 24 items, and is shown to be a valid and reliable version of the ISQ-SR. We conclude that we have achieved equivalence with the original version. PMID- 11026169 TI - K-SADS-PL. PMID- 11026168 TI - The Female Sexuality Questionnaire: instrument development. AB - The purpose of this study was to develop and validate a measure of female sexuality, the Female Sexuality Questionnaire (FSQ). A survey design was employed for the study. The sample included 262 adult women ages 18 to 81 (M = 32). Five hundred questionnaires were distributed to a diverse female population representing multiple geographic areas in the U.S. and Canada. Participants were healthy (as per self-report), sexually active and engaged in an intimate relationship. The majority of the sample was premenopausal. Item stems for the FSQ were developed based on the literature and the domain-referenced approach for item generation. A pool of 70 items generated to represent 6 theoretical domains of female sexuality: Reproduction, Beliefs, Intimacy, Body Image, Physical Responsiveness, and Satisfaction were subjected to systematic review by a panel of experts. A 5-point self-report response scale was appended to the 54 items surviving judges' reviews (criterion validity index of 91.3 for the pool of retained items). Data screening and item analysis was performed and yielded 13 items which were removed prior to principal factor analysis (PFA) resulting in a sample of 218 subjects and 41 items. The PFA with quartrimax rotation resulted in a 5-factor solution. Four of the five were consistent with the hypothesized domains of Satisfaction, Physical Responsiveness, Beliefs, and Body Image. The fifth factor suggested an association with Intimacy, and was renamed Relationships. The hypothesized domain of Reproduction was not supported. Twelve items failed to load on any of the factors. Recommendations for alternations of the final version of the FSQ are based upon the empirical results of the item analysis and PFA. PMID- 11026170 TI - Severe leukopenia with valproate. PMID- 11026172 TI - Eating disorder and superior mesenteric artery syndrome. PMID- 11026171 TI - Trazodone for symptoms of frontal lobe atrophy. PMID- 11026173 TI - Role of the family in the onset and outcome of childhood disorders: selected research findings. AB - OBJECTIVE: Three areas of basic family research were selected for review as being of special importance to the clinically oriented child and adolescent psychiatrist: behavioral genetics, expressed emotion, and the interaction of family dynamics and childhood illness. METHOD: Medline and PSYCINFO searches using appropriate keywords were obtained for each of the 3 major areas. All English-language articles published after 1989 that included empirical research pertaining to children or adolescents were reviewed. RESULTS: Behavioral genetics research indicates that the shared environment, including issues of parental monitoring and discipline, is important in the development and outcome of externalizing disorders. Differential parental treatments of one sibling are critical in internalizing disorders. Criticism (as measured by expressed emotion) is associated with poor outcome of many childhood medical and psychiatric disorders. Chronic illness in a child changes the family dynamics toward being more structured and less emotionally warm and communicative. The family's role in adherence to treatment is critical, and families with high levels of criticism have more difficulty. CONCLUSIONS: Families can cause problems, but many times the problems families have are in response to a child's problems. There is a continued need to empirically assess which family processes are important for specific childhood disorders. PMID- 11026174 TI - Compliance with recommendations to remove firearms in families participating in a clinical trial for adolescent depression. AB - OBJECTIVE: To assess the rate and correlates of compliance with clinicians' recommendations to remove firearms from the homes of depressed adolescents participating in a clinical trial. METHOD: The parents of 106 adolescents with major depression who participated in a randomized psychotherapy clinical trial were asked systematically about firearms in the home. Those who answered affirmatively were given information about the suicide risk conveyed by guns in the home and urged to remove them. The rates of gun removal and acquisition were assessed at the end of the treatment and over the subsequent 2-year naturalistic follow-up. RESULTS: Of those who had guns at intake, 26.9% reported removing them by the end of the acute trial. Retention was associated with urban origin, marital dissatisfaction, and paternal psychopathology. Of those who did not have guns at intake, 17.1% reported acquiring them over 2-year follow-up. Living in a 2-parent household and marital dissatisfaction were associated with gun acquisition. CONCLUSIONS: Families of depressed adolescents may frequently be noncompliant with recommendations to remove guns from the home despite compliance with other aspects of treatment. More efficacious interventions to reduce access to guns in the homes of at-risk youths are needed. PMID- 11026175 TI - Commentary: When counseling parents on guns doesn't work: why don't they get it? PMID- 11026176 TI - Behavioral sequelae of perinatal insults and early family adversity at 8 years of age. AB - OBJECTIVE: This prospective longitudinal study investigated the simultaneous impact of early biological and psychosocial risk factors on behavioral outcome at school age. METHOD: A cohort of 362 children born between 1986 and 1988 with different biological (perinatal insults) and psychosocial risk factors (family adversity) was followed from birth to school age. When their children were aged 8 years, parents of 89.0% of the initial sample completed the Child Behavior Checklist (CBCL). RESULTS: More externalizing as well as internalizing problems were found in children born into adverse family backgrounds, whereas no differences at broad-band syndrome level were apparent between groups with varying obstetric complications. Children with family risk factors had higher scores on 5 of the 8 CBCL scales (including attention, delinquent, and aggressive problems), whereas children with perinatal risk factors had more social and attention problems than children in the nonrisk groups. With one exception, no interactions between risk factors emerged, indicating that perinatal and family risk factors contributed independently to outcome. The differences between risk groups applied irrespective of gender. CONCLUSIONS: The adverse impact of family adversity clearly outweighed the influence of obstetric complications in determining behavioral adjustment at school age. PMID- 11026178 TI - Prodromal symptoms before onset of manic-depressive disorder suggested by first hospital admission histories. AB - OBJECTIVE: A priority for research on manic-depressive or bipolar I disorder (BPI) for children and adolescents has been to search for early predictors of the illness. METHOD: Medical record data were reviewed and systematically coded for a sample of 58 adult patients (32 males/26 females) with confirmed diagnoses of BPI to identify prodromal features and possible patterns of symptoms from the Amish Study. RESULTS: The most frequently reported symptoms included episodic changes in mood (depressed and irritable) and energy plus anger dyscontrol, with no significant gender differences. A progression of ages is seen for the most commonly reported symptoms prior to age 16. The time interval was 9 to 12 years between appearance of the first symptoms and onset of a documented BPI syndrome. CONCLUSIONS: The data suggest testable hypotheses about specific symptoms and behaviors that may be useful for the early detection of children at highest risk for developing manic-depressive disorder. PMID- 11026177 TI - Early childhood heart rate does not predict externalizing behavior problems at age 7 years. AB - OBJECTIVE: In previous research, low resting heart rate in childhood and adolescence has been shown to predict aggressive and/or delinquent behavior at subsequent ages. It has been found that heart rate recorded as early as age 3 years could predict externalizing behavior at age 11 years. This study explored the possibility of a similar relationship between heart rate and externalizing behavior problems. METHOD: Heart rate recorded at ages 14, 20, 24, 36 months and 7 years was used to predict combined parental ratings on the Aggressive and Delinquent Behavior scales of the Child Behavior Checklist (CBCL/4-18) as well as the Externalizing composite scale measured at age 7 years. Subjects consisted of same-sex twin pairs, treated as singletons in the present study, participating in the MacArthur Longitudinal Twin Study. Subjects were grouped into high and low heart rate groups and also into high CBCL/4-18 scoring and low CBCL/4-18 scoring groups. RESULTS: Heart rate was not significantly related to scores on either of the 2 subscales or the Externalizing composite scale at any age. Heart rate group membership did not predict CBCL/4-18 scores. Conversely, CBCL/4-18 group membership did not predict heart rate at any age. CONCLUSION: In this sample, heart rate does not predict externalizing behavior at age 7. PMID- 11026179 TI - Atypical symptoms of depression in a sample of depressed child and adolescent outpatients. AB - OBJECTIVE: To examine the presence of symptoms of atypical depression among children and adolescents with a major depressive disorder (MDD). METHOD: One thousand forty-six youths (aged 6-19 years) meeting DSM-III-R criteria for MDD were included in the study. All subjects had presented at an outpatient clinic seeking treatment and were identified as having MDD via clinical interviews using the semistructured Schedule for Affective Disorders and Schizophrenia for School Age Children-Present Episode (K-SADS-P) with the youngster themselves and a parent/guardian. A diagnosis of atypical depression was derived from the symptoms of depression assessed in the K-SADS-P and required the presence of mood reactivity and at least one the following symptoms: hypersomnia, increased appetite, weight gain, or psychomotor retardation (substituted for leaden paralysis). RESULTS: One hundred sixty-two (15.5%) of the depressed youths met criteria for atypical depression. The symptoms of atypical depression were found to correlate marginally, and the diagnosis of atypical depression had marginal construct validity for both children and adolescents. CONCLUSIONS: The findings from this large sample of depressed children and adolescents suggest that atypical features of depression occur in this age group. However, the diagnosis of atypical depression appears to have only marginal construct validity for both children and adolescents. PMID- 11026181 TI - Explosive outbursts in children with Tourette's disorder. AB - OBJECTIVE: Sudden, explosive outbursts of behavior occur in some children with Tourette's disorder (TD). The etiology of these symptoms is unknown. This study investigated the relationship between explosive outbursts, TD, and its comorbid disorders. METHOD: Tic type and severity and the presence of specific comorbid disorders were compared in 37 children with TD and explosive outbursts and 31 children with TD who did not have such symptoms. RESULTS: Children with TD and explosive outbursts were more likely to demonstrate significant comorbid conditions, particularly attention-deficit/hyperactivity disorder, obsessive compulsive disorder, and oppositional defiant disorder. Tic type and severity did not appear related to the presence of explosive outbursts. A highly significant relationship was demonstrated between the number of comorbid psychiatric diagnoses and explosive outbursts. CONCLUSIONS: Explosive outbursts in children with TD resemble intermittent explosive disorder and may reflect dysregulation of diverse domains of brain function. The presence of such symptoms should alert the clinician to underlying comorbid conditions. PMID- 11026180 TI - Self-esteem in special education children with ADHD: relationship to disorder characteristics and medication use. AB - OBJECTIVES: To describe the level of self-esteem among the study population, to examine how self-esteem ratings may vary by disorder characteristics and medication use, and to identify predictors of low self-esteem while adjusting for sociodemographic factors. METHOD: In a school district-wide sample of children in special education programs, the authors assessed self-esteem with the Piers Harris Self-Concept Scale among 143 students at high risk for attention deficit/hyperactivity disorder (ADHD) in the school year 1995, with an interview participation rate of 73%. Disorder characteristics and medication use were assessed using multiple informants and standardized measures. RESULTS: Self esteem scores, on average, were in the normal range. However, across ADHD comorbidity profiles, children with ADHD and internalizing symptoms had significantly lower self-esteem scores, especially in the areas of anxiety and popularity, than children with ADHD alone or those with comorbid disruptive behavior disorders. Independent predictors of lower self-esteem were being while, high functional impairment, and comorbid internalizing conditions, but not medication use. CONCLUSIONS: Findings suggest that interventions for ADHD should be culturally sensitive as well as aimed at improving a child's functional level and associated internalizing symptoms. Medication use among this younger patient group was not related to self-esteem scores. PMID- 11026182 TI - Relationship of childhood sexual abuse and eating disturbance in children. AB - OBJECTIVE: To test the hypothesis that childhood sexual abuse increases the risk of eating disturbance in children. METHOD: Data obtained from 20 sexually abused children were compared with data obtained from 20 nonabused control children. All subjects were female and between the ages of 10 and 15 years. Subjects completed a battery of tests assessing eating disorder behaviors, body image concern, and childhood trauma history. RESULTS: Comparison of the 2 groups revealed that the abused children had higher levels of weight dissatisfaction and purging and dieting behavior. Furthermore, abused children reported eating less than control children when they felt emotionally upset. Abused children were less likely than control children to exhibit perfectionistic tendencies, but more likely to desire thinner body types. CONCLUSIONS: This is the first controlled study to examine the relationship between childhood sexual abuse and eating disturbance which relied on children as subjects. The results support previous findings with adult subjects which indicate that a history of childhood sexual abuse is associated with weight and body dissatisfaction, along with purging and dietary restriction. PMID- 11026183 TI - Epidemiology and natural course of eating disorders in young women from adolescence to young adulthood. AB - OBJECTIVES: To describe the epidemiology of eating disorders (ED) in a community sample of adolescent girls; to compare the clinical characteristics of full syndrome (FS) and partial-syndrome (PS) ED cases; and to provide information about the continuity between adolescent ED and young adult psychopathology. METHOD: A randomly selected sample of high school girls were assessed during adolescence (n = 891) and a year later (n = 810), and a stratified subset (n = 538) was assessed during their 24th year. The assessments included the Schedule for Affective Disorders and Schizophrenia for School-Age Children, the Longitudinal Interval Follow-up Evaluation, level of functioning, mental health treatment utilization, history of suicide attempt, and physical symptoms. RESULTS: The incidence of ED was less than 2.8% by age 18, and 1.3% for ages 19 through 23. Comorbidity with other psychopathology (89.5%), but especially depression, was very high. FS- and PS-ED groups differed significantly from a no disorder comparison group on most outcome measures, and more than 70% of the adolescent FS- and PS-ED cases met criteria for an Axis I disorder in young adulthood. CONCLUSIONS: FS- and PS-ED are associated with substantial comorbidity, treatment seeking, impaired functioning, and risk for psychopathology in young adulthood. PMID- 11026184 TI - Risk factors for homelessness in adolescents released from psychiatric residential treatment. AB - OBJECTIVES: To measure the incidence of homelessness among youths discharged from an inpatient facility as well as to identify factors that place these youths at risk for homelessness. METHOD: Reports of homelessness during 5 years of follow up interviews with 83 adolescents consecutively discharged from an inpatient facility were used in conjunction with chart reviews. RESULTS: One third of youths experienced at least one homeless episode after discharge. Youths who had a history of substance use, physical abuse, running away, or being in state custody were more likely to become homeless than those without such factors. Having a thought disorder, however, was inversely related to becoming homeless after hospital discharge. CONCLUSIONS: Youths discharged from a residential treatment facility are at high risk for becoming homeless. Interventions to prevent homelessness among adolescent inpatients should be targeted to those with a history of substance abuse, physical abuse, running away, and being in state custody. PMID- 11026185 TI - Initial validation of the functional impairment scale for children and adolescents. AB - OBJECTIVE: Two studies assessed the validity of the Functional Impairment Scale for Children and Adolescents (FISCA), a multidimensional parent-report questionnaire. METHOD: In study 1, quasi-exploratory and confirmatory procedures tested whether FISCA data for 804 inpatients (mean age = 13.4, 456 boys), collected October 1994 through December 1995, fit a 3-factor model. Study 2 (n = 330) used survival and discriminant analyses to predict recidivism status at 3 and 6 months follow-up from FISCA scores at intake. RESULTS: The 8 FISCA scales reduced to 3 factors describing undercontrolled aggression, social role violations, and self-focused aspects of child functional impairment. Serious impairment on the Aggression and School scales each predicted a 3-fold increase in recidivism risk. Together, impairment scores for Aggression, School, Thinking, and Delinquency correctly identified 73% of the recidivists. However, 51% of the nonrecidivists also were classified as recidivists. CONCLUSIONS: Although these data initially support the FISCA's validity, they underscore the need for more effective strategies to identify severely impaired children whose problems will be sporadic or short-lived. PMID- 11026186 TI - Case series: brief parent-child group therapy for childhood anxiety disorders using a manual-based cognitive-behavioral technique. AB - OBJECTIVE: To report on a brief parent-child group therapy program for children with anxiety disorders. METHOD: Twenty-four children with an anxiety disorder and their parents participated in a 10-session treatment. Children were evaluated at pretreatment (T1), posttreatment (T2), 12-month follow-up (T3), and 36-month follow-up (T4). Ten children were also assessed on entering a waiting period (T0). RESULTS: There were no significant symptomatic changes between T0 and T1. Anxiety symptoms decreased significantly during the treatment and follow-up periods. Depressive symptoms changed only during the follow-up period. The percentage of children with no current anxiety disorder was 71% at T2 and 91% at T4. Children of mothers with an anxiety disorder improved more than children of nonanxious mothers, whereas the anxiety level of anxious mothers remained stable. CONCLUSIONS: Brief parent-child group psychotherapy may serve as a time-limited, cost-effective, and efficient intervention. PMID- 11026187 TI - An open trial of plasma exchange in childhood-onset obsessive-compulsive disorder without poststreptococcal exacerbations. AB - Patients with childhood-onset obsessive-compulsive disorder (OCD) with symptom exacerbations following streptococcal infections benefit from treatment with plasma exchange. In this study, 5 patients with treatment-refractory OCD without a history of streptococcus-related exacerbations underwent an open 2-week course of therapeutic plasma exchange. Behavioral ratings, completed at baseline and 4 weeks after the initial treatment, included the Clinical Global Impressions Scale and the Yale-Brown Obsessive Compulsive Scale. All 5 patients completed the trial with few side effects, but none showed significant improvement. Plasma exchange does not benefit children and adolescents with OCD who do not have streptococcus related exacerbations. PMID- 11026188 TI - Heroin use among adolescents in treatment for substance use disorders. AB - OBJECTIVE: To clarify prevalence rates and describe patterns of adolescent heroin users who are in treatment for substance use disorders. METHOD: The Treatment Episode Data Set (TEDS) was examined for trends in the number of adolescents admitted to substance abuse treatment centers and for changes in the routes of heroin administration. Thirteen adolescents who used heroin from one treatment program were compared with 536 adolescents who did not. RESULTS: Between 1992 and 1996, heroin-using youths represented 2.0% of youths in treatment and in 1997 they represented 2.6%. Heroin-using youths represented 56% of those using injection drugs. Heroin-using youths from one treatment program had significantly more polysubstance dependence in comparison with adolescents who did not use heroin. CONCLUSIONS: Nationally, there has been an increasing number, but not percentage, of heroin-using youths in treatment between 1992 and 1996. In 1997 there was an increase in both the number and percentage of heroin-using youths in treatment. Heroin-using adolescents have the highest rate of injection drug use when compared with youths using other substances. Because of their greater risk of contracting human immunodeficiency virus through injection drug use, treatment trials for these adolescents are needed. PMID- 11026189 TI - Contingency management reinforcement procedures for adolescent substance abuse. PMID- 11026190 TI - Single-subject designs. PMID- 11026191 TI - Genetics of childhood disorders: XIX. ADHD, Part 3: Is ADHD a noradrenergic disorder? PMID- 11026192 TI - [Laparoscopic cholecystectomy in choledocholithiasis and stricture of the terminal part of the common bile duct]. AB - The authors offer the treatment and diagnostic algorithm in choledocholithiasis, stricture of a terminal portion of the common hepatic duct and papilla stenosis revealed in laparoscopic cholecystectomy (LCE). With the purpose of intraoperative assessment of bile ducts states during LCE, the diagnostic system including laparoscopic and ultrasonic examinations, cholangiography and choledochoscopy was developed and applied. In intraoperative revealing of choledocholithiasis without bile outflow disorders and wide cystic duct the authors prefer to remove the concrements during choledochoscopy through cystic duct without intervention on Vater's papilla (VP). In combination of choledocholithiasis with bile outflow disorders and also in isolated papilla stenosis and stricture of a terminal portion of the common hepatic duct, one stage laparoscopic cholecystectomy, intraoperative antegrade papillosphincterotomy and retrograde calculus extraction is optimal. In cases when complete endoscopic calculus extraction is impossible, the drainage of the common hepatic duct by Cholsted's with subsequent delayed endoscopic papillosphincterotomy (EPST) is acceptable. During intraoperative examination in 49 patients (57.6%) the concrements in choledochus, not diagnosed earlier, were revealed, in 21--stricture of terminal choledochus portion and in 19 patients- papilla stenosis. In 12 cases the concrements were removed during choledochoscopy through the cystic duct stump, 4 patients with big concrements required laparoscopic choledocholithotomy. In 16 cases LCE with various variants of choledochus drainage was performed as the first stage, as the second stage--EPST and lithoextraction. Antegrade papillosphincterotomy was performed in 15 patients during LCE. In 12 cases intraoperatively revealed choledocholithiasis combined with papilla stenosis (7) and choledochus stricture (5) was the indications to intraoperative papillosphincterotomy. Papilla stenosis was the indication to antegrade papillosphincterotomy in 3 patients. PMID- 11026193 TI - [Intraperitoneal perfusion of perftoran in the treatment of patients with diffuse purulent peritonitis]. AB - For higher efficacy of general purulent peritonitis treatment, intraperitoneal perfusion of Perftoran emulsion was proposed. The drug was administered as a single dose of 100-150 ml before wound suturing. In 48 hours an increase of phagocytizing cells fraction in peritoneal exudate and their higher functional activity was revealed: the phagocytic activity and phagocytic index was 2 times higher than in the control group, complete phagocytosis index--1.5 times higher. Significant reduction of microbial contamination in abdominal cavity was also revealed: in 48 hours there were less aerobic bacteria than in the control group, there were no anaerobic bacteria, or anaerobes were detected in a minimum quantity. Local use of Perftoran allowed to decrease the rate of repeated peritoneal sanitation by 44.4% and to decrease mortality by 5.2%. PMID- 11026194 TI - [Correction of the intraintestinal status in patients with diffuse peritonitis]. AB - Complex methods of intraintestinal status correction are proposed for patients surgically treated for general peritonitis. These methods consist of three main components: suppression of excessive bacterial colonization of gastrointestinal tract; binding of toxic substances and microorganisms; colonization of small intestine by physiological strains of bacteria. Original design of three-lumen tube for nasogastrointestinal intubation was used. Clinical study was carried out in 2 groups of patients. Study group consisted of 79 patients treated by proposed methods; only gastrointestinal intubation by three-lumen tube was performed in 44 patients (control group). Usage of the proposed methods led to dysbacteriosis correction and fast appearance of intestinal peristalsis, improved the course of postoperative period. The mortality was 8.9% in the study group and 13.6% in the control group. PMID- 11026195 TI - [Importance of the morphofunctional status of APUD cells in the prognosis of duodenal ulcer hemorrhage]. AB - In 24 patients with uncomplicated duodenal ulcer and 40 patients with acute bleeding duodenal ulcer morphofunctional status of the local paraendocrine adjusting system of the duodenal mucous membrane in the periulcer, ulcer and remote zones was studied with histochemical and electron-microscopic techniques. It is established, that ECL- and G-apudocytes hyperplasia and hyperfunction and vice versa D-cells reduction of the number and suppression of the secretory activity are reliable prognostic criteria of duodenal ulcer bleeding. PMID- 11026196 TI - [Abdominal trauma with injuries of the major vessels]. AB - Treatment results in 87 patients (mean age 30.1 years) with traumatic injuries of major abdominal vessels were analyzed. 73 patients had stab-incised wounds, 9- blunt trauma, 5--gunshot wounds. Hemodynamics of the majority of hospitalized patients (66 from 87) was unstable. Inferior cava vein injury was revealed in 29 patients, aorta injury--in 21, iliac vessels injury--in 22, visceral vessels injury--in 15. One vascular wall was injured in 58 cases, two walls--in 27 cases, complete transection or separation of vessel was revealed in 6 cases. Major vessel injuries were associated with abdominal visceral trauma in 97% cases. Vascular operations (side suture--in 49 patients, circular suture--in 11, vein ligation--in 5, prosthesis--in 3) were performed in 17 (25%) patients by general surgeons, in the rest (75%)--by vascular surgeons. General mortality (including hospitalized but non-operated patients, and patients who died during the operation and in postoperative period) was 46%. It is necessary to teach vascular surgery to general surgeons for high-quality and timely care in patients with abdominal trauma complicated by major vessels injuries. PMID- 11026197 TI - [Diagnosis of heart contusion in associated chest trauma]. AB - The efficacy of different methods of heart contusion diagnosis in patients with concomitant chest trauma was studied. Criteria of heart contusion diagnosis by electrocardiography, integral body rheography, ultrasonic diagnosis, heart enzymes analysis were covered. Comparative analysis revealed that there was no modern method with 100% diagnostic value. Only complex use of these methods permits to make objective diagnosis of heart contusion. This approach allowed to decrease diagnostic mistakes rate in patients with concomitant chest trauma by 18%. PMID- 11026198 TI - [Diagnostic laparoscopy in pregnant women]. AB - 54 laparoscopic examinations were performed in 52 pregnant women when acute surgical diseases of the abdomen were suspected. When the suspicions of acute surgical diseases in pregnant women are substantiated, diagnostic laparoscopy has high diagnostic potential and is safe for the mother and fetus. This method permits to avoid unnecessary open operations, and also to perform simple operations through laparoscope. PMID- 11026199 TI - [Noninvasive methods of the examination in surgery of obliterating atherosclerosis of lower extremity arteries]. AB - The purpose of the study was to determine surgical policy in atherosclerotic lesions of aorta and lower extremities arteries on the basis on non-invasive diagnostic methods--duplex scanning (DS) and computer tomographic angiography (CTA). From 1998 to 1999 47 patients were examined on this program (DS + CTA). At the first stage of the study, 17 patients underwent translumbal aorto arteriography at the same time with DS and CTA. Analysis of the angiograms revealed that sensitivity of angiography for detection of lesions in aorto-iliac segment was 94%, in femoro-popliteal segment--78%. Diagnostic mistakes were associated with partial thrombosis of aneurysm's lumen, as a result of which the aorta looked non-dilated in angiograms. Femoral arteries contrasted unsatisfactory in 4 patients because of low collateral blood velocity distal to occlusion of iliac arteries. Initial segments of deep and superficial femoral arteries closed each other in angiograms on anterior-posterior projection, and it also led to low potential of angiography in assessment of femoral arteries state. Complex of non-invasive diagnostic methods (DS + CTA) allowed to correct diagnosis in each case. The methods complemented one another: CTA permitted to make three-dimensional reconstruction of vascular tree and to study its on various projections; DS--to study the vascular wall's state and hemodynamical parameters of blood flow. At the second stage of the study, high sensitivity of DS and CTA allowed to reject traditional angiography in preoperative period. Intraoperative findings confirmed the provisional diagnosis completely. PMID- 11026200 TI - [Prevention of early thrombotic complications after reconstructive operations in occlusions of the aorto-iliac segment]. AB - 108 patients with atherosclerosis of aorto-iliac segment have undergone various reconstructive operations on this segment. Depending on the treatment all the patients were divided in 2 groups (test group--54 patients, control group--54 patients). The patients of the control group received aspirin, rheopolyglucin and trental for correction of hemorheology and hemocoagulation disorders. In patients of the test group at the same time as this treatment, the indirect electrochemical blood oxidation by intravenous infusion of 0.03% sodium hypochlorite was applied. Regional hemodynamics, hemorheology and hemostasis indices were studied in all the patients. It was established that indirect electrochemical blood oxidation permits to significantly improve the hemorheology indices, to diminish thrombophilia phenomenon during the treatment and early postoperative period. It was noted that early thrombotic reocclusion and amputations rates were reduced in the test group. PMID- 11026201 TI - [Fiber fasciotomy in Dupuytren's contracture]. AB - 42 patients with hand Dupuytren's contracture were operated. Transection of each finger scar cords was performed in transverse direction through 3-8 skin punctures from proximal portion to distal segment in the zone of middle phalanx base. In addition, complete finger extension was achieved on operating table. There were no recurrences in follow-up of 13 years. PMID- 11026202 TI - [Incidence of urination disorders and erectile dysfunction after surgical treatment of congenital anorectal defects in children]. AB - For assessment of the incidence and characteristics of urination disorders and erectile dysfunction after radical surgical treatment of anorectal congenital defects in children, 60 patients (12 girls, 48 boys) without urination disorders before operation, who have undergone 1-4 proctoplasties were examined. Their was Hirschsprung's disease in 20 patients, anorectal atresia in 40 patients. Iatrogenic urination disorders and erectile dysfunction were revealed in 38 (63.3%) patients, majority of them (24) underwent repeated proctoplasty. The main urination disorders were urinary incontinence (30 patients), infravesical obstruction (10), hyporeflexion of urinary bladder. Erectile dysfunction after operation was in 11 (34.4%) patients from 32. The combination of disorders was revealed in 17 (44.7%) patients. Primary radical operations in children with anorectal defects often cause iatrogenic urination disorders and erectile dysfunction. The number of complications increases progressively after repeated proctoplasty. It is necessary to improve the treatment and the methods of surgical correction bearing in mind the risk of iatrogenic disorders of genitourinary apparatus. PMID- 11026203 TI - [Treatment strategy in diverticulosis of the colon]. AB - The results of treatment of 645 patients with diverticular disease of the colon have been analysed. There were 65.4% of women and 34.6%--of men, 54.4% of patients were hospitalized urgently and 45.6% electively. 82.5% of patients underwent conservative treatment, 24.8% of them were operated. In hyperkinetic type of bowel motor activity preference was given to mini-invasive organ-saving operations. In 28 cases Laparoscopic serosa myotomy by Reilly and Hodson were carried out (in 17 cases through miniapproach). In hypokinetic type various types of resection of the colon (20 patients) were performed. In necessity for urgent operation various methods of operations may be made depending on general condition of the patient and extension of the pathologic process. In 29 cases initial resection with anastomosis has been performed. Postoperative complications developed in 20.5% patients. There were no lethal outcomes. In 66 cases 2-stage operations were carried out, lethality being 10.6%, and complications--23.7%. PMID- 11026204 TI - [Indirect electrochemical oxidation of blood plasma in combined treatment of patients with acute mediastinitis]. AB - Treatment results in 61 patients with acute mediastinitis are analyzed. Combined treatment of acute mediastinitis, including cryoplasma-antienzymic complex use and indirect electrochemical blood plasma detoxication, promotes disappearance of microcirculation disorders, to earlier decreasing of endogenic intoxication, outcomes improvement. PMID- 11026206 TI - [Comment on the article by Ia.P. Kulik and S.N. Pokoliukhin "Time to make choice: preventive or curative appendectomy" (a viewpoint)]. PMID- 11026205 TI - [Experience in the use of imipenem/cilastatin (thienam) in a surgical clinic]. PMID- 11026207 TI - [Reconstructive surgery of lower extremity arteries: revascularization through the system of deep arteries of the thigh]. PMID- 11026208 TI - [Photodynamic therapy in suppurative diseases of soft tissues]. PMID- 11026209 TI - Testing legal assumptions regarding the effects of dancer nudity and proximity to patron on erotic expression. AB - A field experiment was conducted in order to test the assumptions by the Supreme Court in Barnes v. Glen Theatre, Inc. (1991) and the Ninth Circuit Court of Appeals in Colacurcio v. City of Kent (1999) that government restrictions on dancer nudity and dancer-patron proximity do not affect the content of messages conveyed by erotic dancers. A field experiment was conducted in which dancer nudity (nude vs. partial clothing) and dancer-patron proximity (4 feet; 6 in.; 6 in. plus touch) were manipulated under controlled conditions in an adult night club. After male patrons viewed the dances, they completed questionnaires assessing affective states and reception of erotic, relational intimacy, and social messages. Contrary to the assumptions of the courts, the results showed that the content of messages conveyed by the dancers was significantly altered by restrictions placed on dancer nudity and dancer-patron proximity. These findings are interpreted in terms of social psychological responses to nudity and communication theories of nonverbal behavior. The legal implications of rejecting the assumptions made by the courts in light of the findings of this study are discussed. Finally, suggestions are made for future research. PMID- 11026210 TI - Police responses to sexual assault complaints: the role of perpetrator/complainant intoxication. AB - This study explored the impact of victim and perpetrator alcohol consumption on police officers' evaluations of an alleged sexual assault and their reported likelihood of charging the perpetrator. Two hundred and twelve police officers were presented with a vignette depicting an acquaintance rape in which the beverage consumption (beer, cola) of both the victim and perpetrator was systematically varied. Results indicated that the officers' perceptions of the complainant's intoxication level, as well as the gender of the officer, influenced officers' evaluations of the alleged sexual assault. The more intoxicated the complainant was perceived to be, the more negatively she was viewed. Female police officers evaluated the victim more favorably than male officers. The only factors related to the officers' likelihood of charging the perpetrator, however, involved their assessment of the complainant's credibility and their perception of the likelihood that the perpetrator would be found guilty in a court of law. PMID- 11026211 TI - Distributive and procedural justice in seven nations. AB - The paper examines the impact of distributive justice and procedural justice variables on judgments in seven countries (Bulgaria, France, Hungary, Poland, Russia, Spain, and the United States). Subjects were presented with each of two experimental vignettes: one in which the actor unsuccessfully appeals being fired from his job and one in which the actor unsuccessfully goes to an employment agency to seek a job; they were asked to rate the justness of the outcome and how fairly the actor had been treated. The vignettes manipulated deservingness and need of the actor (distributive justice factors) and impartiality and voice in the hearing (procedural justice factors). Four hypotheses were tested: first, a distributive justice hypothesis that deservingness would be more important than need in these settings; second, a procedural justice hypothesis that the importance of voice and impartiality vary depending on the nature of the encounter and the forum in which it is resolved; third, because of their recent socialist experience, Central and Eastern European respondents make greater use of need information and less use of deservingness information than Western respondents; and fourth, that distributive justice and procedural justice factors interact. The distributive justice hypothesis is supported in both vignettes. The procedural justice hypothesis receives some support. Impartiality is more important in the first vignette and voice is more important in the second vignette. The interaction hypothesis was not supported in the first vignette, but does receive some support in the second vignette. The cultural hypothesis is not supported in either vignette. The implications for distributive and procedural justice research are discussed. PMID- 11026212 TI - The perceived validity of eyewitness identification testimony: a test of the five Biggers criteria. AB - The U.S. Supreme Court has outlined five criteria on which evaluations of eyewitness identifications should be based (certainty, view, attention, description, and time; Neil v. Biggers, 1972). We postulated that certainty plays a qualitatively different role from the four other Biggers criteria in evaluations of eyewitness identification testimony. Specifically, we hypothesized that participants would ignore reports on other criteria when certainty was high (the certainty-trumps hypothesis), but not when certainty was low. Participants (N = 386) read a fictitious trial transcript in which three of the five Biggers criteria were manipulated (certainty, view, and attention, or certainty, description, and time) and completed a questionnaire. The certainty-trumps hypothesis was not supported. Instead, the Biggers criteria combined only as main effects, not interactions, supporting a summative hypothesis. Surprisingly, collateral effects indicated that manipulations of one criterion (e.g., certainty) affected perceptions of other criteria (e.g., attention and view) and vice versa. Implications of the results are discussed. PMID- 11026213 TI - The uncritical acceptance of risk assessment in forensic practice. AB - Forensic psychologists are frequently asked to conduction evaluations of risk assessment. While risk assessment has considerable merit, recent applications to forensic psychology raise concerns about whether these evaluations are thorough and balanced. Forensic adult risk-assessment models stress risk factors, and deemphasize or disregard entirely the other side of the equation: protective factors. Mediating and moderating effects must also be considered. Moreover, base rate estimates may produce erroneous results if applied imprudently to forensic samples without regard to their unstable prevalence rates or the far-reaching effects of settings, referral questions, and evaluation procedures. Psychologists are offered a preliminary list of relevant issues for evaluating the merits of risk assessment in their forensic practices. PMID- 11026214 TI - [Cardiologists discover a heart for chocolate. Infarct prevention with chocolate segments?]. PMID- 11026215 TI - [Renewed discussion on calcium antagonists in hypertension. More myocardial infarcts?]. PMID- 11026216 TI - [New hope for difficult to treat corneal damage. Cornea from the laboratory. Interview by Petra Eiden]. PMID- 11026217 TI - [Is nature fighting back? Environmental poisons as reproduction inhibitors]. AB - Environmental noxae may impair male fertility. Different compartments and types of cell must be considered as possible targets for drugs, such lifestyle factors as alcohol and smoking, and various environmental and occupational agents. However, the mechanisms of action in humans are known for only a limited number of noxae. For the clinical andrologist, the identification of relevant exposure to reproductive toxicants by careful history-taking remains of great importance. Nevertheless, evaluation of the relevant risks in the individual is often not possible. PMID- 11026218 TI - [Male infertility. Current life style could be responsible for infertility]. AB - Optimal spermatogenesis requires the testicles to be at a lower temperature than the body core. This is achieved by the following factors:--Blood in the testicular artery is precooled by the surrounding veins of the plexus pampiniformis; nevertheless, high fever results in substantial warming of the testicles;--Heat loss via the scrotal skin, with tight-fitting, thermally insulating clothing or obesity having an unfavorable effect;--increased circulation of air around the genitals on physical activity;--High temperature gradient to the environment when ambient temperatures are low. If the combination of these factors is unfavorable, disturbed spermatogenesis and fertility may result, which, however, is usually reversible. Likewise, electromagnetic waves may impair spermatogenesis by heat induction in the testicles, but only when exposure is excessive. PMID- 11026219 TI - [Sperm intolerance. "My partner has an allergic reaction to me"]. PMID- 11026220 TI - [Diabetic foot syndrome series, 3. Basic principles in treatment]. PMID- 11026221 TI - [Increasing the dose, evaluating compliance, revising the diagnosis. Prescriptions for apparently resistant processes in the esophagus]. AB - One of the most common reasons for failed treatment of esophageal diseases is inadequate medication dosage. This applies in particular to gastro-esophageal reflux disease (GERD). In many cases, however, the adoption of unrealistic therapeutic aims or inappropriate target criteria may play a role. Last, but not least, adequate compliance on the part of the patient is a prerequisite for successful treatment. In any case of resistance to treatment, it is recommended that the diagnosis be checked, and the algorithms on how to proceed issued by the specialist associations (re-)consulted. PMID- 11026222 TI - [Arthroses. 1: Classification, clinical aspects and differential diagnosis]. PMID- 11026223 TI - [Diagnostic quiz. Stomach pain with a long history. Double pylorus]. PMID- 11026224 TI - [Soon one "female distress" less. Irritable bowel syndrome responds to serotonin blockers]. PMID- 11026225 TI - [Pneumococcal infections. You can protect your patients]. PMID- 11026226 TI - [Treatment in general practice. Effective, well tolerated therapy]. PMID- 11026227 TI - [Experiences with a needleless injection system. Immunotherapy without needle fear]. PMID- 11026228 TI - [Hemodynamics]. PMID- 11026229 TI - [Regulation of the circulation system]. PMID- 11026230 TI - [An outline of neural regulation of blood pressure]. PMID- 11026231 TI - [Cardiovascular regulation by baroreceptor reflex and vasomotor center]. PMID- 11026232 TI - [Central regulation of blood pressure by renin-angiotensin system in the brain]. PMID- 11026233 TI - [Blood pressure regulation by nitric oxide within the brain]. PMID- 11026234 TI - [Role of brain L-arginine in central regulation of blood pressure]. PMID- 11026235 TI - [Simulation of cardiovascular control system]. PMID- 11026236 TI - [Brief review: molecular biology of vasoactive substance and signal transduction]. PMID- 11026237 TI - [The renin-angiotensin system and its receptors]. PMID- 11026238 TI - [Tissue renin angiotensin system]. PMID- 11026239 TI - [Endothelin]. PMID- 11026240 TI - [Vasopressin]. PMID- 11026241 TI - [Neuropeptide Y]. PMID- 11026242 TI - [Noradrenaline, adrenaline]. PMID- 11026243 TI - [Subtypes and function of serotonin]. PMID- 11026244 TI - [Insulin]. PMID- 11026245 TI - [Vasoactive substances: prostaglandins]. PMID- 11026246 TI - [Arachidonic acid metabolites (HETEs and EETs) produced by cytochrome P450s]. PMID- 11026247 TI - [Leukotriene]. PMID- 11026248 TI - [Dopamine system]. PMID- 11026249 TI - [Adrenomedullin and related peptides]. PMID- 11026250 TI - [PTH/PTH-related peptide]. PMID- 11026251 TI - [Calcitonin gene-related peptide]. PMID- 11026252 TI - [Kallikrein-kinin systems]. PMID- 11026253 TI - [The natriuretic peptide system]. PMID- 11026254 TI - [Nitric oxide]. PMID- 11026255 TI - [Steroid hormone]. PMID- 11026256 TI - [Molecular medicine in leptin biology]. PMID- 11026257 TI - [Glucagon]. PMID- 11026258 TI - [Hepatocyte growth factor (HGF)]. PMID- 11026259 TI - [Endogenous digitalis-like factors]. PMID- 11026260 TI - [Endothelium-derived hyperpolarizing factor]. PMID- 11026261 TI - [L-arginine]. PMID- 11026262 TI - [HO (heme oxygenase)/CO (carbon monoxide) system]. PMID- 11026263 TI - [Hypertension and reactive oxygen species]. PMID- 11026264 TI - [VEGF and vasculoangiogenesis/vascular remodeling]. PMID- 11026265 TI - [Signal transduction of JAK-STAT pathway]. PMID- 11026266 TI - [Oxidized low-density lipoprotein receptors]. PMID- 11026267 TI - [Significance of PPAR gamma(peroxisome proliferator-activated receptor gamma) in hypertension and its vascular complications]. PMID- 11026269 TI - [Soluble guanylyl cyclase]. PMID- 11026268 TI - [Hypertension-associated activation of Ras.MAP kinase system]. PMID- 11026270 TI - [Definition, classification and severity of hypertension]. PMID- 11026271 TI - [Methods of blood pressure measurement and comparison with each method]. PMID- 11026272 TI - [A clinical significance and current problems of home blood pressure measurements]. PMID- 11026273 TI - [Ambulatory blood pressure monitoring in hypertensive patients]. PMID- 11026274 TI - [The sixth report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure, and 1999 World Health Organization-International Society of Hypertension Guidelines for the Management of Hypertension]. PMID- 11026275 TI - [Genetic aspect of hypertension]. PMID- 11026276 TI - [Role of central nervous system and sympathetic nervous system in essential hypertension]. PMID- 11026277 TI - [Cerebral hemodynamics in essential hypertension]. PMID- 11026278 TI - [Renal hemodynamics in essential hypertension]. PMID- 11026279 TI - [Neural control of renal hemodynamics in essential hypertension]. PMID- 11026280 TI - [Control of renal hemodynamics by cytochrome P450-dependent metabolites of arachidonic acid]. PMID- 11026281 TI - [Brain microcirculation]. PMID- 11026282 TI - [Coronary microcirculation]. PMID- 11026283 TI - [Renal microcirculation]. PMID- 11026284 TI - [Neuronal control of mesenteric microcirculation]. PMID- 11026285 TI - [Shear stress]. PMID- 11026286 TI - [Vascular endothelium and vascular smooth muscle cells]. PMID- 11026287 TI - [Peripheral hemodynamics]. PMID- 11026288 TI - [Vascular remodeling]. PMID- 11026289 TI - [Sodium and potassium metabolism]. PMID- 11026290 TI - [Metabolism of calcium and magnesium]. PMID- 11026291 TI - [Ion channel and hypertension]. PMID- 11026292 TI - [Sodium transport systems in red cell membrane]. PMID- 11026293 TI - [Alterations in vascular smooth muscle sodium transport in hypertension]. PMID- 11026294 TI - [Sodium transport in renal tubule]. PMID- 11026295 TI - [Na(+)-Ca2+ exchange, Ca(2+)-ATPase]. PMID- 11026296 TI - [Ca and Na regulation in cardiac cells]. PMID- 11026297 TI - [Sodium sensitivity in essential hypertension]. PMID- 11026298 TI - [Sleep and diurnal change in blood pressure]. PMID- 11026299 TI - [Blood pressure change induced by meal]. PMID- 11026300 TI - [Bathing, relating behaviors and blood pressure fluctuation]. PMID- 11026301 TI - [Nocturnal blood pressure (dipper, non-dipper) and morning blood pressure]. PMID- 11026302 TI - [Circadian variation of blood pressure in shift workers]. PMID- 11026303 TI - [Time structures of blood pressure variability]. PMID- 11026304 TI - [Catecholamine]. PMID- 11026305 TI - [Dopamine receptor in hypertension]. PMID- 11026306 TI - [Serotonin in cardiovascular disease]. PMID- 11026307 TI - [The renin-angiotensin system]. PMID- 11026308 TI - [Vasopressin]. PMID- 11026309 TI - [Prostaglandins and their related substances]. PMID- 11026310 TI - [Kallikrein-kinin in hypertension]. PMID- 11026311 TI - [Endothelin receptor antagonist]. PMID- 11026312 TI - [Nitric oxide]. PMID- 11026313 TI - [Natriuretic peptide family]. PMID- 11026314 TI - [Insulin]. PMID- 11026315 TI - [Pathophysiological roles of PTH, PTHrP, and CGRP in hypertension]. PMID- 11026316 TI - [Behavior and pathophysiological significance of adrenomedullin in essential hypertension]. PMID- 11026317 TI - [Steroid hormone]. PMID- 11026319 TI - [Genetics of essential hypertension]. PMID- 11026318 TI - [Roles of endogenous digitalis-like factors in hypertension]. PMID- 11026320 TI - [Renin gene]. PMID- 11026321 TI - [Angiotensinogen gene]. PMID- 11026322 TI - [Angiotensin-converting enzyme gene]. PMID- 11026323 TI - [Angiotensin II type 1 receptor gene]. PMID- 11026324 TI - [Alpha-adducin gene]. PMID- 11026325 TI - [Glucocorticoid receptor gene]. PMID- 11026326 TI - [Sa gene]. PMID- 11026327 TI - [Insulin receptor gene and hypertension]. PMID- 11026328 TI - [LDL receptor]. PMID- 11026329 TI - [alpha-Adrenergic receptor gene]. PMID- 11026330 TI - [beta-Adrenergic receptor gene]. PMID- 11026331 TI - [Ob gene]. PMID- 11026332 TI - [CYP11B2, CYP17]. PMID- 11026333 TI - [klotho gene]. PMID- 11026334 TI - [G protein genes]. PMID- 11026335 TI - [Endothelial nitric oxide synthase gene and hypertension]. PMID- 11026336 TI - [Gene of atrial natriuretic peptide]. PMID- 11026337 TI - [Gene expression of neuropeptide Y]. PMID- 11026338 TI - [Human renal kallikrein (hKLK1) gene]. PMID- 11026339 TI - [Other candidate genes]. PMID- 11026340 TI - [Epidemiology of hypertension in Japanese]. PMID- 11026341 TI - [Ethnic differences in the genetic background of essential hypertension]. PMID- 11026342 TI - [Geographical pathology of hypertensive diseases--epidemiological studies on 60 populations in the world]. PMID- 11026343 TI - [The changing pattern of hypertension in a long-term follow-up survey of a Japanese community: the Hisayama Study]. PMID- 11026344 TI - [Ohasama Study--a longitudinal cohort study for hypertension]. PMID- 11026345 TI - [Population-based cohort studies of cardiovascular disease in Japan]. PMID- 11026346 TI - [Prospective studies of cardiovascular disease in Japanese work site population]. PMID- 11026347 TI - [Distribution of blood pressure in university students in Japan: cardiovascular risk factors disclosed from the annual examination of health]. PMID- 11026348 TI - [Possibility of diagnosis by genes]. PMID- 11026349 TI - [Aging and blood pressure]. PMID- 11026350 TI - [Predictor of hypertension]. PMID- 11026351 TI - [Natural history]. PMID- 11026352 TI - [Blood pressure variation--an overview]. PMID- 11026353 TI - [Effects of alcohol intake on blood pressure]. PMID- 11026354 TI - [Acute and chronic effects of smoking on blood pressure]. PMID- 11026355 TI - [Cold stimuli and cold pressor test]. PMID- 11026356 TI - [Psychological stress]. PMID- 11026357 TI - [Exercise stress test]. PMID- 11026358 TI - [Effects of sodium, potassium, calcium, and magnesium intakes in hypertension]. PMID- 11026359 TI - [Effect of caffeine and other drinks containing caffeine on blood pressure]. PMID- 11026360 TI - [Hypertension and vitamin]. PMID- 11026361 TI - [Seasonal variation of blood pressure]. PMID- 11026362 TI - [Salt sensitivity in essential hypertension]. PMID- 11026363 TI - [The risks of hypertension]. PMID- 11026364 TI - [Hypertension and cognitive function]. PMID- 11026365 TI - [Cerebrovascular diseases]. PMID- 11026366 TI - [Hypertensive cardiac disease]. PMID- 11026367 TI - [Electrocardiographic findings]. PMID- 11026368 TI - [Hypertension and QT dispersion]. PMID- 11026369 TI - [Echocardiographic findings]. PMID- 11026370 TI - [Prognostic significance of quantification of left ventricular mass in hypertensive patients]. PMID- 11026371 TI - [Molecular mechanisms of cardiac hypertrophy induced by high blood pressure]. PMID- 11026372 TI - [Hypertensive hypertrophic cardiomyopathy]. PMID- 11026373 TI - [Arrhythmias]. PMID- 11026374 TI - [Ischemic heart disease in hypertensive patients]. PMID- 11026375 TI - [Heart failure]. PMID- 11026376 TI - [Renal damage due to hypertension: an overview]. PMID- 11026377 TI - [Pathogenesis of renal damage due to hypertension]. PMID- 11026378 TI - [Urinary abnormalities in patients with hypertension]. PMID- 11026379 TI - [Hypertension and atherosclerosis]. PMID- 11026380 TI - [Diagnosis and clinical relevance of carotid atherosclerosis as a hypertensive vascular damage]. PMID- 11026381 TI - [Hypertensive retinopathy]. PMID- 11026382 TI - [Malignant hypertension]. PMID- 11026383 TI - [Hypertension and sudden death]. PMID- 11026384 TI - A simple and powerful test for autocorrelated errors in OLS intervention models. AB - The important assumption of independent errors should be evaluated routinely in the application of interrupted time-series regression models. The two most frequently recommended tests of this assumption [Mood's runs test and the Durbin Watson (D-W) bounds test] have several weaknesses. The former has poor small sample Type I error performance and the latter has the bothersome property that results are often declared to be "inconclusive." The test proposed in this article is simple to compute (special software is not required), there is no inconclusive region, an exact p-value is provided, and it has good Type I error and power properties relative to competing procedures. It is shown that these desirable properties hold when design matrices of a specified form are used to model the response variable. A Monte Carlo evaluation of the method, including comparisons with other tests (viz., runs, D-W bounds, and D-W beta), and examples of application are provided. PMID- 11026385 TI - Conservation skills among preschool children in Umtata, South Africa. AB - To 41 boys and 29 girls (M age = 5.2 yr.) in a South African preschool a test of conservation of liquid was given. Analysis showed conservation skills had not been acquired as only 5 children showed such skills and only one knew why. PMID- 11026386 TI - Preferences and expectations of innovator-ritualist types in relation to sex of university students in a Turkish sample. AB - The aim of this study was whether there is a prevailing representation of Innovators versus Ritualists which differs in both expected frequency and preference of the types for the two sexes. 213 Turkish undergraduate students were given Ritualist and Innovator vignettes in a random order and asked to state the expected frequency of the vignettes if each described a male or female. In addition to this, they stated their preferences for each sex in vignettes. Analysis showed that women are expected more frequently to be Ritualist than men, and men are expected to be more frequently Innovators than women. Moreover, regardless of sex, people prefer Innovators to Ritualists, while they state that they expect to encounter Ritualists more frequently than Innovators in Turkish society. Research implications and suggestions for study are discussed. PMID- 11026387 TI - The factor structure of the Minnesota Satisfaction Questionnaire and participants of vocational rehabilitation. AB - This study investigated the factor structure of the Minnesota Satisfaction Questionnaire when used by individuals with psychiatric disabilities who are participating in vocational rehabilitation. The sample consisted of 87 respondents recruited from Columbus, Ohio, in 1996 who worked in noncompetitive employment. Factor analyses with varimax rotation conducted on the short-form of the questionnaire indicated three factors, an intrinsic factor and an extrinsic factor (as proposed by the Herzberg two-factor theory) as well as another pertaining to satisfaction derived from participating in vocational rehabilitation. PMID- 11026388 TI - Detecting latent taxa: Monte Carlo comparison of taxometric, mixture model, and clustering procedures. AB - A Monte Carlo evaluation of four procedures for detecting taxonicity was conducted using artificial data sets that were either taxonic or nontaxonic. The data sets were analyzed using two of Meehl's taxometric procedures, MAXCOV and MAMBAC, Ward's method for cluster analysis in concert with the cubic clustering criterion and a latent variable mixture modeling technique. Performance of the taxometric procedures and latent variable mixture modeling were clearly superior to that of cluster analysis in detecting taxonicity. Applied researchers are urged to select from the better procedures and to perform consistency tests. PMID- 11026389 TI - Three last letters identify most female first names. AB - The last spelled letter often indicates the sex of first names. Most female names end with the last letter a, e, or i. Female names are distinctive and therefore easy to identify because of the small number of predominantly female endings. Most male names end with one of 19 last letters. Two last letters, h and y, occur with similar frequency for both sexes. The cultural tradition of higher status for men contributes to the avoidance of predominantly female last letters for male names. A female name with a predominantly male last letter therefore occurs more often than a male name with a predominantly female last letter. The findings were obtained from the 500 most frequent first names given to males and females in Pennsylvania in 1990. PMID- 11026390 TI - Job insecurity in a sample of Canadian civil servants as a function of personality and perceived job characteristics. AB - Economic downturns and organisational changes have stimulated studies on the importance of job security for public employees; however, there has been some disagreement as to whether job insecurity should be defined using a single- or a two-factor model, on how to measure it and which antecedents and consequences are linked to it. Questionnaires measuring job insecurity as well as personality traits and job characteristics as possible antecedent variables, and job performance and job satisfaction as possible consequences were completed by a sample of 71 federal government employees during recent important government downsizing. A multiple regression analysis indicated that job insecurity, defined as a combination of organisational or personal vulnerability, was significantly related to only two antecedent variables, i.e., Neuroticism and job characteristics. Further regression analyses indicated that job insecurity was a statistically significant moderator but only between one consequence, i.e., intention to quit, and the job characteristics variable. These results along with methodological issues and further research, are discussed. PMID- 11026391 TI - Personality, job level, job stressors, and their interaction as predictors of coping behavior. AB - Hypotheses concerning the relationships among job stressors, job level, personality, and coping responses were investigated in a sample of 305 electrical contracting employees. Coping behaviors were measured with questionnaire items based on interviews conducted with a sample of the subjects. Neuroticism (N) and Extraversion (E) were the personality variables most strongly related to coping behavior. Overall, more coping variance was explained by personality than by job stressors; however, when the effects of job level and job stressors were combined, they explained more variance in complaining/quitting and seeking social support than did the personality variables. Both work situation and personality seem to be important variables in the choice of coping behaviors. There was no evidence of interactions among personality, stressors, and job level in explaining coping behavior. PMID- 11026392 TI - Unwillingness to communicate and satisfaction in organizational relationships. AB - To examine the relationship between unwillingness to communicate and quality of organizational relationships 200 employees of several organizations completed the Unwillingness to Communicate Scale and the Organizational Communication Relationships subscale of the International Communication Association Audit. Correlations indicated that people who reported increased communication and high reward in communication also reported greater relational satisfaction and greater perceived organizational influence. PMID- 11026393 TI - Illusion of control relative to chance outcomes. AB - Participants were engaged in a computer game in which outcomes occurred independently of responses. Depending upon group assignment, positive outcomes occurred randomly on either 33% or 66% of trials. Outcome relative to chance was manipulated by varying the number of response alternatives from which participants could choose (two, three, or six). After 50 and 100 trials, participants rated the control they had over accruing points and were asked to suggest specific rules as to how points were obtained. Illusion of control was related to percentage of trials producing favorable outcomes but was insensitive to performance relative to chance. Thus, participants ignored basic statistical probabilities in assessing their control, relying solely on the likelihood of positive outcomes. Furthermore, illusion of control was not related to participants' scores on Burger and Cooper's 1979 Desirability of Control scale. PMID- 11026394 TI - Rated satisfaction with salary and promotion by secondary school teachers in Transkei. AB - Among 114 secondary school teachers there was no significant difference between the 55 who were satisfied with their salary and the 59 who were not, but the difference was significant between 37 who were satisfied with promotion and 73 who were not. There was no sex difference for either measure. PMID- 11026395 TI - Predicting involuntary dismissal for unauthorized absence, lateness, and poor performance in the selection of unskilled and semi-skilled British contract factory operatives: the contribution of the Employee Reliability Inventory. AB - This report contains the results of a predictive validity study, which examined the relationship between the employment status of a group of temporary contract operatives in a British factory, 90 days after being hired (N = 50) and their scores on the Employee Reliability Inventory. Scores were systematically related to employment status. Operatives who were involuntarily dismissed by their employer for unauthorized absence, lateness, or poor performance had poorer scores than those who remained on the job. Some preliminary benefit-cost implications are discussed. PMID- 11026396 TI - Glucose levels are not associated with common features-derived prototype memory abstractions. AB - Elevated glucose levels are associated with increased recall on declarative memory tasks. No previous studies have examined such a relationship using a common features-derived prototype memory abstraction task. 20 participants volunteered, 10 each in an experimental group who were given 40 g of glucose to drink and a control group given an artificially sweetened beverage after fasting. Analysis indicated that common features-derived prototype memory abstractions seem resistant to glucose fluctuations. PMID- 11026397 TI - MMPI-2 profiles of Colombian, Mexican, and Venezuelan university students. AB - Performance of 82 Colombian, 87 Mexican, and 96 Venezuelan university students and adult community dwellers on Spanish translations of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) were compared. Findings suggest more comprehensive research is required on applications of the MMPI-2 in Latin America and greater consideration of the roles of culture and nationality. PMID- 11026398 TI - College students' responses to content-specific advisories regarding television and movies. AB - Extending Ingold's research on adult viewers' responses to mature content advisories, this study examined the likelihood of viewing television programs and of viewing theatrical movies with general and content-specific advisories. The results show that among the 63 undergraduate men, viewing likelihood was increased significantly with the provision of all types of mature content advisories, whereas among the 122 undergraduate women viewing interest increased with the 'sexuality' advisory but decreased with the 'violence' advisory. PMID- 11026399 TI - Adolescents' meaning in life: a replication of DeVogler and Ebersole (1983). AB - Previous studies have indicated that adolescents understand the concept of meaning in life and can apply it to their lives. In this study we examined how religious adolescents differ in their self-reports of meaning in life compared to adolescents previously studied. It is expected that adolescents in a Christian community more frequently attribute their meaning in life to beliefs than will adolescents in the non-Christian population. 81 adolescents from a religious summer camp reported commitment to a belief as their strongest personal meaning more frequently than any other category when reported in a free-response essay format, whereas no adolescents in the previous study reported belief as their strongest meaning. PMID- 11026400 TI - Comparison of 26-item and 20-item versions of the Toronto Alexithymia Scale for psychiatric outpatients. AB - This study compared the 20-item revised Toronto Alexithymia Scale with the 26 item version for 257 psychiatric outpatients. Scores on the 20-item version significantly correlated with those on the 26-item inventory even when controlled for depressed mood. Furthermore, the 20-item scale showed greater internal consistency. Factor analysis for both versions indicated three subfactors but the factors in the TAS-20 accounted for greater common variance (92%) than for the factors in the TAS-26 (78%). The total variance accounted for by these factors on the TAS-20 (45%) was also greater than for the TAS-26 (38%). PMID- 11026401 TI - Preschoolers' knowledge about strategies in coping with other's disgust evoked by immoral actions. AB - 40 preschoolers in Exp. 1 and 22 in Exp. 2 (mean ages were both 5:11) were shown short stories presented as colored videotaped pictures with explanatory narrations. In each story a recipient felt disgusted by an agent's action. In Exp. 1 the agent's action was immoral. The participants were asked to tell how the agent would behave, supposing they were the agent themselves. About 80% of their answers were prosocial. In Exp. 2, two kinds of story were shown. In one, the agent hurt the recipient intentionally; in the other, by accident. Almost all answers in both kinds of story were prosocial. Furthermore, over a third of the participants told the reasons for their answers, considering the recipient's emotion, even when the agent's action was intentional and immoral. These findings show that the preschoolers had suitable knowledge about the agent's strategies in coping with the recipient's disgust. PMID- 11026402 TI - Methods to decrease attrition in longitudinal studies with adolescents. AB - A summary of methods to decrease attrition in longitudinal school-based studies conducted with adolescents beginning junior high schools or middle schools is presented. These include collection of contact information about students, additional days to collect data from absentee students, data collection in new high schools once students graduate from junior high schools or middle schools, sending questionnaires by mail, and conducting telephone or home interviews. PMID- 11026403 TI - Type A behavior pattern, sex, and social support. AB - Low social support has been suggested as a contributing factor to cardiac disease in some individuals with Type A Behavior Pattern. A number of studies have focused on the relationship between social support and Type A traits in subjects without cardiac disease as a means of evaluating this hypothesis; however, results of these studies have been confusing. In the current study, involving samples of adolescents and working adults, there was a significant positive correlation between Type A scores and satisfaction with social support in men but not in women. PMID- 11026404 TI - Cheating among University of Transkei students. AB - The purpose of this study was to investigate the extent to which cheating in academic work occurs at this university. 58 participants registered for a postgraduate diploma in education responded to a questionnaire on cheating. Some students admitted engaging in every behaviour; however, the percentage of students so engaged was much lower than those reported among British and American institutions of higher learning. This may partly be attributed to respondents being reluctant to divulge their dishonesty. PMID- 11026405 TI - Differences between Caucasian and Hispanic undergraduates in emphasis on distributive, procedural, and interactional justice. AB - Most theories of justice focus on individual-level approaches; few examine the role of contextual factors such as culture. To fill this gap in the literature, this study examined the differences between Caucasian and Hispanic undergraduates (n = 120 and 58, respectively) in terms of their emphasis on distributive, procedural, and interactional justice. Analyses indicated that Hispanic students were more likely to use interactional justice than Caucasian respondents, but the groups did not differ in their likelihood of using distributive or procedural justice. PMID- 11026406 TI - Meaningfulness as a mediator of subjective well-being. AB - In a sample of 347 university students and community residents, measures of meaningfulness, self-esteem, internal locus of control, positive social relationships and optimism were given to predict subjective well-being, i.e., measures of happiness, life satisfaction, and affect balance. Correlations, canonical correlations, and structural equation modeling supported the hypothesis that meaningfulness is a significant mediator between personality variables and subjective well-being. PMID- 11026407 TI - Sociomoral reflection and prosocial and antisocial behavior: two Italian studies. AB - Two Italian studies on the development of moral judgment based on the 1992 Sociomoral Reflection Measure-Short Form of Gibbs, Basinger, and Fuller were performed. In Study 1, 49 male and female adolescent volunteers involved in prosocial activities and a control group of 60 nonvolunteers completed the form. In Study 2, 60 male drug abusers in a drug-rehabilitation community were compared with a matched group of 60 nondrug abusers. Analysis showed that involvement in prosocial activities was associated with higher stages of moral judgment and Type B responses, while drug abuse was associated with lower stages of moral judgment and more Type A responses. The moral type scores were less closely associated with behavioral outcomes than the moral stage scores, perhaps reflecting the interplay between universal sociocognitive processes and specific real-life situations. PMID- 11026408 TI - Personality profiles of environmentally responsible groups. AB - The hypothesis that groups high and low on environmental responsibility have different personality profiles was tested using responses of 319 introductory psychology students (132 men and 187 women) who completed the ECOSCALE and the Revised NEO Personality Inventory for partial course credit. Results of discriminant function analysis supported the hypothesis, indicating that groups scoring high and low on Environmental Responsibility had significantly different personality profiles and that the standard discriminant function coefficients were quite substantial (> or = .50) for Openness (.72) and Agreeableness (.50). PMID- 11026409 TI - Voluntary class absences and other behaviors in college students: an exploratory analysis. AB - In a survey of 120 sophomore and junior undergraduate college students, we found that those who admitted to cutting class for no reason during the previous month had a lower mean GPA than those who reported no voluntary absences. Cutting class was related to reports of getting drunk, speeding, breaking the law, and visiting home, although reports of speeding, getting drunk, and breaking the law were not related to GPA. The 11 students who reported feeling depressed had a lower GPA than those without depression, while the 18 students who had not visited their families in the previous month had higher GPAs than those who had visited their families. Implications for scale development are discussed. PMID- 11026410 TI - Comparison of locus of control among Muslim and Jewish school students in Iran. AB - A comparison was made of scores on locus of control by 210 Muslim and 210 Jewish school students. These two groups were matched on sex, age, and father's education. No significant differences were found between Muslim and Jewish students. Also, frequencies of the responses to each scale item in two religious groups were compared. The results are considered in relation to previous studies and some cultural implications are addressed. PMID- 11026411 TI - Ego strength and stress reaction in Kuwaiti students after the Iraqi invasion. AB - Correlations between scores on the Barron Ego Strength Scale and a checklist of 18 Symptoms of stress reactions were calculated for two responses from 503 undergraduate Kuwaiti students (225 men and 278 women). Using total scores on both measures for all students the correlation was -.42, so students with higher ego strength scores tend to have fewer stressful symptoms. rs by symptoms ranged from -.33 to -.11. PMID- 11026412 TI - Internal consistency reliability and validity of the Hebrew translation of the Oxford Happiness Inventory. AB - The Hebrew translation of the Oxford Happiness Inventory and the short form Revised Eysenck Personality Questionnaire were completed by 298 undergraduate women in Israel. The findings confirm the internal reliability of the Hebrew translation of the Oxford Happiness Inventory and support the construct validity according to which "happiness is a thing called stable extraversion." PMID- 11026413 TI - Diagnosing written language disabilities using the Woodcock-Johnson Tests of Educational Achievement-Revised and the Wechsler Individual Achievement Test. AB - The writing portions of the Woodcock-Johnson Tests of Educational Achievement Revised and the Wechsler Individual Achievement Test are often administered when establishing eligibility for special education services due to learning disabilities. The scores on these measures are typically regarded as equivalent although little is known about how scores on the two measures differ for the same students. Differences of only a few points, however, may affect eligibility for special education services. These tests were administered to 25 sixth grade students previously diagnosed with learning disabilities in written expression only. Students' Wechsler scores were consistently higher on the overall writing composite, while there was no difference in the mean scores on the language mechanics subtests. The WIAT Written Expression subtest mean, however, was significantly higher than the Woodcock-Johnson Writing Samples subtest mean. Use of the Wechsler test would be less likely to identify children for special education services in written expression when point discrepancy criteria are utilized for eligibility. Clinicians should be cognizant of the effect of the specific test chosen on eligibility outcome. PMID- 11026414 TI - Psychosocial, behavioral, and biometric characteristics of stages of exercise adoption. AB - This study incorporates the Stages of Change model to examine the relationship between the stages of exercise adoption and indicators of health and fitness. Subjects were 198 United States Forest Service (USFS) workers recruited from 8 USFS ranger stations in the Pacific Northwest. Analysis indicated that those subjects in Action and Maintenance stages of exercise adoption differed significantly from those in Preparation and Precontemplation/Contemplation stages on a variety of indicators of both health and fitness. Also, interesting differences were noted among the physically inactive stages, e.g., Preparation stage subjects showed lower triglycerides, systolic blood pressure, and higher HDL cholesterol compared with subjects in the Precontemplation/Contemplation stage. Progressing to the Preparation stage, where the individual remains inactive but is aware of the problem and has begun to make small changes, is associated with a differing (more positive) health profile in this study compared to those in the Precontemplation/Contemplation stage. These results imply that simply advancing in stage can have a variety of health benefits. PMID- 11026416 TI - Effect on tipping of barman drawing a sun on the bottom of customers' checks. AB - Previous research has demonstrated that a pleasant drawing (a smiling face) on a restaurant bill increased the number of tips left by clients. A similar experiment was carried out using a drawing of the sun since it is known that tips increase on sunny days. The experiment was carried out in local bars and involved clients who have ordered an espresso coffee. Analysis showed that the drawing of the sun led clients to leave a tip more frequently than when this drawing is not present. The size of the tip left was also higher. The hypothesis of the creation of a positive frame of mind by this stimulus is discussed. PMID- 11026415 TI - Stress and immune system function in a newspaper's newsroom. AB - Data were collected on 10 newswriters in a newspaper's newsroom. These included self-reported stress ratings and saliva samples for secretary immunoglobulin A (IgA) analysis. The stress ratings and saliva samples were taken initially and after three 30-min. periods. The first time period was a baseline measure and involved the workers engaging in their usual tasks. At Time 2 music was presented for 30 min. while workers engaged in their normal activities. At Time 3 workers resumed their normal duties. Findings showed an increase in IgA that was not statistically significant, a reduction in stress during the music period that was statistically significant, and a statistically significant negative correlation between stress and IgA. PMID- 11026417 TI - Relations among positive affect, negative affect, and somatic symptoms in a medically ill patient sample. AB - The Positive and Negative Affect Scale (PANAS), a brief measure of Positive and Negative Affect, may be useful in assessing mood of medical patients because it does not include somatic items frequently confounded with medical conditions. In previous research Positive and Negative Affect have been independent and uncorrelated, and Negative Affect but not Positive Affect has been positively correlated with somatic symptoms. However, relationships between variables may vary in different populations, and there is relatively little information on Positive and Negative Affect in medical patients. In the current study, the PANAS was used to assess the relationships among Positive Affect, Negative Affect, and somatic symptoms and pain in a medically ill hospital population. Positive and Negative Affect scores were positively correlated and for patients reporting pain, Positive Affect scores were positively correlated with pain intensity. Results from this and other studies indicate that Positive and Negative Affect are independent but in some populations may be correlated; positive affects such as hopefulness may co-occur with both somatic complaints and psychological distress. PMID- 11026418 TI - Examiners' errors on the Wechsler Memory Scale-Revised. AB - This study examined the nature of WMS-R scoring errors made by trainee psychologists and the extent to which errors influenced index scores on this scale. Trainee psychologists undertook competency based training exercises for the WMS-R, similar to those devised for the WAIS-R by Fantuzzo, Blakely, and Gorsuch in 1989. A WMS-R training protocol was devised and scored by three experienced psychologists to generate a gold standard protocol. The training protocol was then given to trainee psychologists to score. Scoring errors made by trainees included mathematical and transcription errors, and errors of judgment given failure to apply scoring criteria correctly. Present results are consistent with previous investigations of scoring errors among trainees on Wechsler's intelligence tests, and findings provide a preliminary indication that scoring errors on the WMS-R may alter classification by ability. PMID- 11026419 TI - A follow-up study of infants who wake at night. AB - A retrospective follow-up study of 68 adolescents treated for night waking when infants or toddlers showed that 82% of them were described by their parents as currently sleeping "well," and only two continued to experience a night-waking difficulty. PMID- 11026420 TI - Psychological functioning of recently bereaved, middle-aged women: the first 13 months. AB - This study examined the natural course of psychological functioning in recently bereaved middle-aged women. 69 widows were assessed four times (T1-T4) between the period of 4 to 13 mo. after the loss and were compared to a matched nonwidowed group of 57. Of the SCL-90 feelings of depression, agoraphobic behavior, anxiety, hostility, somatization, feelings of insufficiency, and sleep disorders were heightened at 4 mo. after bereavement compared to the norm group. Significantly higher psychological dysfunctioning was found on all SCL-90 subscales than for non-widows. Over time, a decrease in psychological dysfunction was found for most widows; however, not every widow appeared to recover psychologically, and 17% of the widows showed severe psychological dysfunctioning at 13 mo. postbereavement (T4). With respect to the predictive value of the Total score on the SCL-90, at 13 mo., 27% of these widows had scores indicating severe psychological dysfunctioning; these were comparable to their scores at 4 mo. postbereavement. PMID- 11026421 TI - Written expression, Hooper, and blind rating. AB - Much research has supported the assertion of Hooper, et al.'s 1994 claims that specific stimuli perform better than others in eliciting well-developed written responses; however, previous research was conducted with raters who were aware of the hypotheses. The present study of 29 middle school children validated previous support for Hooper, et al.'s assertions by implementing blind rating and, once again, finding that significant differences between Hooper-like and non-Hooper like prompts existed for structural items but not for items of mechanics. PMID- 11026422 TI - An asymmetric test of homogeneity of proportions. AB - An asymmetrical test of homogeneity of proportions possesses distinct advantages over a symmetrical test. The symmetric chi square test of homogeneity is widely employed in psychological research. An asymmetrical alternative to the chi square test of homogeneity is proposed and described. PMID- 11026423 TI - MMPI-2 scores of Puerto Rican, Mexican, and U.S. Latino college students: a research note. AB - In this brief report, we present MMPI-2 basic validity and clinical scale data of Latino-descent persons from Puerto Rico (n = 290), Mexico (n = 1,920), and the United States (n = 28). All were administered one of three Spanish translations of the MMPI-2. A review of the mean scores of these respective groups indicates similarities across all scales. Differences among these three groups, with the exception of the Mf scale (which is keyed to sex), were well within the one standard deviation band. More importantly, these findings are promising given the fact that three different translations of the MMPI-2 were applied. PMID- 11026424 TI - Psychometric characteristics in normal and social phobic samples for a Spanish version of the Rosenberg Self-Esteem Scale. AB - The present paper had three purposes: (a) presenting normative data for the Rosenberg Self-esteem Scale in a Spanish sample, (b) studying whether there are significant sex or age differences in self-esteem, and (c) studying whether there are significant differences between a Control group with no psychological diagnosis and a group of social phobics. Of the total sample of 266 persons, 214 belonged to the Control group and 52 to the Social Phobic group. Item-total score correlations and alpha reliabilities supported the internal consistency of the scale. There were statistically significant differences between the Control and Social Phobic groups, but not by sex or age. PMID- 11026425 TI - Demographic factors as predictors of coping strategies among police officers. AB - This study examined the effects of demographic factors on coping responses among police officers. A sample of 233 police officers completed the Ways of Coping Questionnaire (Folkman & Lazarus, 1988). The regression analysis showed that the higher the reported educational attainment, the more police officers reported coping which was emotion-focused and seeking social support. The rank of the officer was directly related to reported emotion-focused coping. These results are discussed relative to research on the relations of demographic factors and coping responses among police officers. PMID- 11026426 TI - Validity of the Learning Process Questionnaire with students of lower academic attainment. AB - The Learning Process Questionnaire was administered to 692 secondary school students (308 boys, 380 girls, and 4 unspecified) in the Philippines. The ages of the participants ranged from 14 to 22 years, with an average of 15 yr. An exploratory factor-analysis procedure was conducted for the entire sample, for the lower achievers (n = 333), and for higher achievers (n = 359). The factor structure of the Learning Process Questionnaire was uninterpretable with data obtained from students of lower academic achievement. PMID- 11026427 TI - Students' state motivation and instructors' use of verbally aggressive messages. AB - This study examined the relationship between college students' (107 men, 123 women) state motivation and their instructors' perceived use of 10 verbally aggressive messages, e.g., attacks on competence, character, background, and physical appearance; malediction, teasing, ridicule, threats, swearing, or nonverbal symbols. Significant negative correlations were obtained between students' state motivation and instructors' use of seven verbally aggressive messages: attacks on competence, character, or background, malediction, ridicule, threats, and nonverbal symbols. These findings suggest that these types of verbally aggressive messages are related to students' state motivation whereas attacks on physical appearance, teasing, and swearing by the instructor are not related to students' state motivation. PMID- 11026428 TI - A case study on support for students' thinking through computer-mediated communication. AB - This is a case study on support for thinking through computer-mediated communication. Two graduate students were supervised in their research using computer-mediated communication, which was asynchronous and written; the supervisor was not present. The students' reports pointed out there was more planning and editing and low interactivity in this approach relative to face-to face communication. These attributes were confirmed by their supervisor's report. The students also suggested that the latter was effective in support of a production stage of thinking in research, while the former approach was effective in support of examination of thinking. For distance education to be successful, an appropriate combination of communication media must consider students' thinking stages. Finally, transient and permanent effects should be discriminated in computer-mediated communication. PMID- 11026429 TI - Construct validity of repression: a dimensional analysis. AB - This study explored the construct validity of repression through an examination of the interrelations among indicators identified in previous research as being associated with the construct. Three behavioral tasks, i.e., dichotic listening task, recall of past events task, and Stroop task, modified in accordance with previous research to tap into repressive tendencies, and one questionnaire (Byrne Repression-Sensitization Scale) were administered to a sample of 62 university undergraduates. A series of correlational analyses provided weak to moderate support for the construct validity of repression. PMID- 11026430 TI - MMPI-A performance of African and Mexican American adolescent first-time offenders. AB - 54 African and Mexican American adolescent first-time offenders were examined with the MMPI-A to evaluate ethnic differences. Multivariate analyses by ethnicity and MMPI-A scales (validity, clinical, content, and supplementary scales) were not significant; however, there was a significant univariate difference where African American adolescents scored significantly higher on the Repression scale than the Mexican American group. A greater percentage of within normal-limits profiles were African American (50%) than Mexican American (25%). Research and clinical implications for using the MMPI-A with these groups are discussed. PMID- 11026431 TI - Sex differences in memory estimates for pictures and words. AB - Memory performance estimates of men and women before and after a recall test were investigated. College students (17 men and 20 women), all juniors, participated in a memory task involving the recall of 80 stimuli (40 pictures and 40 words). Before and after the task they were asked to provide estimates of their pre- and postrecall performance. Although no sex differences were found for total correct recall, recall for pictures, and recall for words, or in the estimates of memory performance before the recall task, there were significant differences after the test: women underestimated their performance on the words and men underestimated their performance on the picture items. PMID- 11026432 TI - Differences in immediacy between traditional and nontraditional students. AB - Reported differences between 83 traditional and 44 nontraditional college students on immediacy were nonsignificant. PMID- 11026433 TI - Another failure to generalize the Mozart effect. AB - Several studies have not replicated Rauscher, Shaw, and Ky's 1993 finding that 10 minutes of exposure to Mozart piano music temporarily enhanced performance on three spatial reasoning tasks. Later Rauscher and Shaw argued that enhanced performance is unlikely unless three conditions are met. The present study was designed to meet those three conditions. 36 adults were exposed to one of six listening orders and one of six test orders. Listening and test orders had no systematic effect on spatial reasoning performance. A one-factor, repeated measures analysis of variance yielded no significant difference on spatial reasoning performance after listening to classical music, jazz, or silence. A reanalysis, using only those items most likely to tap spatial reasoning, fell short of significance, and mean scores were in the direction opposite to that hypothesized. These results were inconsistent with studies that have supported a Mozart effect. PMID- 11026434 TI - Is the picture bizarreness effect a generation effect? AB - Bizarre stimuli usually facilitate recall compared to common stimuli. This investigation explored the so-called bizarreness effect in free recall by using 80 simple line drawings of common objects (common vs bizarre). 64 subjects participated with 16 subjects in each group. Half of the subjects received learning instructions and the other half rated the bizarreness of each drawing. Moreover, drawings were presented either alone or with the name of the object under mixed-list encoding conditions. After the free recall task, subjects had to make metamemory judgments about how many items of each format they had seen and recalled. The key result was that a superiority of bizarre pictures over common ones was found in all conditions although performance was better when the pictures were presented alone than with their corresponding label. Subsequent metamemory judgments, however, showed that subjects underestimated the number of bizarre items actually recalled. PMID- 11026435 TI - Phase analysis of burnout and other psychological phenomena. AB - Phase analysis appears to have the potential to increase knowledge and improve ameliorative solutions for burnout. This article explored potential uses of the phase models of burnout and social and practical implications of the phases. The use of phase analysis to increase understanding of other psychological phenomena such as anxiety, citizenship behavior, and hardiness is introduced. PMID- 11026436 TI - AIDS awareness among South African students. AB - The present study explored reported awareness of AIDS among third-year majors in education. The 71 men and 93 women had a mean age of 24.7 yr. (SD = 4.7 yr.). Those who were aware outnumbered those who were not. PMID- 11026437 TI - The effects of being born in the United States on the job satisfaction of Asian Americans. AB - Analysis of the responses of Asian American (n = 178), African American (n = 1,026), and European American (n = 8,118) full-time workers to 21 nationwide surveys representative of the U.S. labor force from 1972 through 1996 showed the job satisfaction of Asian Americans compared to that of the other two groups was affected by whether subjects were born in the United States. In addition, there were no gender differences in job satisfaction among African Americans and European Americans who were and were not born in the U.S., but there were such differences among Asian Americans. PMID- 11026438 TI - Immunization for Alzheimer's disease: a shot in the arm or a whiff? PMID- 11026439 TI - Neurobiology of schizophrenia. AB - Schizophrenia is a common chronic and disabling brain disease of unknown etiology, pathogenesis, and mechanism. Whether schizophrenia represents a single disorder of markedly variable expression or a family of clinically related disorders is unclear. Genetic factors, most likely multiple genes of modest effect, play a major role in its etiology, but an environmental "second hit" may be necessary for clinical expression. The inherited biological susceptibility to schizophrenia is probably expressed clinically as nonpsychotic abnormal personality traits, plus numerous biological markers (cognitive, anatomical, and psychophysiological) that are all found significantly more commonly in the population than is schizophrenia. Neuropathological studies suggest that schizophrenia may be a neurodevelopmental disorder, characterized by reduced neuropil, with no evidence for ongoing cell death. A hypothesized mechanism for these changes involves cell migrational abnormalities occurring in the fetal brain. Schizophrenia is also distinguished by biochemical abnormalities involving the dopamine, GABA, and glutamate systems, and NMDA and nicotinic receptors. Structural and functional brain imaging studies suggest both global and regional abnormalities as well as "disconnections" of specific cerebral circuits. Despite the lack of knowledge regarding pathophysiology, there are reasonably effective treatments for schizophrenia. As the neurobiology of the disorder is unraveled, more effective, targeted treatments will become available. PMID- 11026440 TI - Nasal administration of amyloid-beta peptide decreases cerebral amyloid burden in a mouse model of Alzheimer's disease. AB - Progressive cerebral deposition of amyloid-beta (Abeta) peptide, an early and essential feature of Alzheimer's disease (AD), is accompanied by an inflammatory reaction marked by microgliosis, astrocytosis, and the release of proinflammatory cytokines. Mucosal administration of disease-implicated proteins can induce antigen-specific anti-inflammatory immune responses in mucosal lymphoid tissue which then act systemically. We hypothesized that chronic mucosal administration of Abeta peptide might induce an anti-inflammatory process in AD brain tissue that could beneficially affect the neuropathological findings. To test this hypothesis, we treated PDAPP mice, a transgenic line displaying numerous neuropathological features of AD, between the ages of approximately 5 and approximately 12 months with human Abeta synthetic peptide mucosally each week. We found significant decreases in the cerebral Abeta plaque burden and Abeta42 levels in mice treated intranasally with Abeta peptide versus controls treated with myelin basic protein or left untreated. This lower Abeta burden was associated with decreased local microglial and astrocytic activation, decreased neuritic dystrophy, serum anti-Abeta antibodies of the IgG1 and IgG2b classes, and mononuclear cells in the brain expressing the anti-inflammatory cytokines interleukin-4, interleukin-10, and tumor growth factor-beta. Our results demonstrate that chronic nasal administration of Abeta peptide can induce an immune response to Abeta that decreases cerebral Abeta deposition, suggesting a novel mucosal immunological approach for the treatment and prevention of AD. PMID- 11026441 TI - Epileptogenesis after status epilepticus reflects age- and model-dependent plasticity. AB - Although epilepsy often begins in childhood, factors that contribute to the development of epilepsy as a consequence of status epilepticus (SE) during early development are poorly understood. We investigated animal models in which seizure induced epileptogenicity could be studied. Rats undergoing self-sustaining SE induced by perforant path stimulation (PPS) at the ages of postnatal day 21 (P21) and P35 were compared with those subjected to SE by lithium and pilocarpine (LiPC). Although only one animal subjected to PPS at P21 developed chronic spontaneous seizures by several months of observation, all the animals subjected to PPS at P35 became epileptic. In the LiPC model, however, most of the rat pups subjected to SE at P21 became epileptic. Animals with spontaneous seizures showed increased inhibition in the dentate gyrus, a characteristic of the epileptic brain, with evidence of mossy fiber synaptic reorganization. Examination of circuit recruitment by c-Jun immunohistochemistry showed activation restricted to the hippocampus in P21 animals subjected to PPS, although extensive activation of hippocampal and extrahippocampal structures was seen in pups subjected to PPS induced self-sustaining SE at P35 or LiPC SE at P21. These results demonstrate that the appearance of epilepsy as a consequence of SE is influenced by the type of insult as well as by age-dependent circuit recruitment. PMID- 11026442 TI - Release of the neuronal glycoprotein ICAM-5 in serum after hypoxic-ischemic injury. AB - Intercellular adhesion molecule (ICAM)-5 (telencephalin) is unique among the ICAMs, because it is only expressed in somatodendritic membranes of telencephalic neurons. To investigate the fate of ICAM-5 during focal brain injury, we induced hypoxia-ischemia (HI) damage in adult mice by right common carotid artery ligation followed by hypoxia. ICAM-5 was detectable in serum within a 48-hour window after HI injury. In HI brain, dendritic ICAM-5 immunore-activity was abolished, but it was present in the neuropil and soma of hippocampal pyramidal, dentate granule, and some cortical and striatal neurons. After HI injury, levels of ICAM-5 protein and messenger RNA initially increased, and ICAM-5 messenger RNA expression then decreased, although protein levels continued to increase. Because HI injury induces microglial activation with increases in CD11a/CD18 (lymphocyte function antigen [LFA]-1) counterreceptors to ICAM-5, we investigated whether modulation of interactions between LFA-1 receptors and brain ICAM-5 during HI injury are associated with changes in levels of serum ICAM-5. Intracerebroventricular administration of lipopolysaccharide to activate microglia before HI injury resulted in elevated serum ICAM-5 levels compared with those in mice with only HI injury. Pretreatment with anti-LFA-1 antibodies before HI injury or LFA-1 receptor knockout mice with HI injury had markedly reduced levels of serum ICAM-5. Lipopolysaccharide levels increased, whereas LFA-1 receptor blockade or LFA-1 knockout decreased HI injury in the first 12 hours. These data suggest that during the necrotic phase of HI injury, serum ICAM-5 may be a potential marker for somatodendritic neuronal damage. PMID- 11026443 TI - Normalization of elevated CD4-/CD8- (double-negative) T cells after thymectomy parallels clinical remission in myasthenia gravis associated with thymic hyperplasia but not thymoma. AB - T-cell-dependent B-cell help is likely to be of major importance in the pathogenesis of myasthenia gravis, but mechanisms provoking a pathological anti acetylcholine receptor (AChR) response are poorly understood. We report on the dysregulation of recently identified CD4-/CD8- (double-negative) T cells (DN T cells), which have been shown to participate in immunoregulation and antibody augmentation. Compared with healthy controls, significantly increased frequencies of DN T cells were found in the blood of myasthenia gravis patients with lymphofollicular hyperplasia. After thymectomy, however, normalization in the number of these cells was seen in parallel with clinical improvement and reduction in anti-AChR antibody titers. The effect of thymectomy was observed irrespective of adjuvant treatment and held true for up to 4 years of follow-up. In marked contrast, frequencies similar to control values were found in myasthenia gravis patients with thymoma, with thymectomy having no further reducing effect. These data indicate that CD4-/CD8- T cells not only participate in the pathogenesis of myasthenia gravis but also correlate with disease activity and histological findings. PMID- 11026444 TI - Dopa-responsive dystonia is induced by a dominant-negative mechanism. AB - Dopa-responsive dystonia (DRD) is induced by a deficiency of GTP cyclohydrolase I (GCH) and has a postulated autosomal dominant inheritance with a low penetrance. G201E is a dominant DRD mutation. Recombinant G201E mutant protein possessed very low enzyme activity. When G201E was expressed in eukaryotic cells, only a small amount of GCH protein could be detected. In baby hamster kidney cells, G201E protein was synthesized normally but was degraded rapidly in pulse-chase experiments. More interestingly, G201E dramatically decreased the level of wild type protein and GCH activity in cotransfection studies. Therefore, G201E exerts a dominant-negative effect on the wild-type protein, probably going through an interaction between them. We also showed that L79P but not R249S (a recessive DRD mutation) had a dominant-negative effect. Through the dominant-negative mechanism, a single mutation could decrease GCH activity to less than 50% of normal. This study not only explains the inheritance of DRD but also increases the understanding of genetic diseases associated with multiple subunit proteins. PMID- 11026445 TI - The role of the insular cortex in temporal lobe epilepsy. AB - The role of the insular cortex in the genesis of temporal lobe epileptic (TLE) seizures has been investigated in 21 patients with drug-refractory TLE using chronic depth stereotactic recordings of the insular cortex activity and video recordings of ictal symptoms during 81 spontaneous electroclinical seizures. All of the recorded seizures were found to invade the insula, most often after a relay in the ipsilateral hippocampus (19/21 patients). However, 2 patients had seizures that originated in the insular cortex itself. Ictal symptoms associated with the insular discharges were similar to those usually attributed to mesial temporal lobe seizures, so that scalp video-electroencephalographic monitoring does not permit making any difference between ictal symptoms of temporo-mesial and insular discharges. A favorable outcome was obtained after a temporal cortectomy sparing the insular cortex in 15 of 17 operated patients. Seizures propagating to the insular cortex were found to be fully controlled by surgery, whereas those originating in the insular cortex persisted after temporal cortectomy. The fact that seizures originating in the insular cortex are not influenced by temporal lobectomy is likely to explain some of the failures of this surgical procedure in TLE. PMID- 11026446 TI - Axonal Guillain-Barre syndrome: relation to anti-ganglioside antibodies and Campylobacter jejuni infection in Japan. AB - To clarify the relations of the axonal form of Guillain-Barre syndrome (GBS) to anti-ganglioside antibodies and Campylobacter jejuni infection, 86 consecutive Japanese GBS patients were studied. Electrodiagnostic criteria showed acute inflammatory demyelinating polyneuropathy in 36% of the patients and acute motor axonal neuropathy (AMAN) in 38%. Frequent anti-ganglioside antibodies were of the IgG class and against GM1 (40%), GD1a (30%), GalNAc-GD1a (17%), and GD1b (21%). Identified infections were C. jejuni (23%), cytomegalovirus (10%), Mycoplasma pneumoniae (6%), and Epstein-Barr virus (3%). There was a strong association between AMAN and IgG antibodies against GM1, GD1a, GalNAc-GD1a, or GD1b. Almost all the patients with at least one of these antibodies had the AMAN pattern or rapid resolution of conduction slowing/block possibly because of early-reversible changes on the axolemma. C. jejuni infection was frequently associated with AMAN or anti-ganglioside antibodies, but more than half of the patients with AMAN or anti-ganglioside antibodies were C. jejuni-negative. These findings suggest that the three phenomena "axonal dysfunctions (AMAN or early-reversible conduction failure)," "IgG antibodies against GM1, GD1a, GalNAc-GD1a, or GD1b," and "C. jejuni infection" are closely associated but that microorganisms other than C. jejuni frequently trigger an anti-ganglioside response and elicit axonal GBS. PMID- 11026447 TI - Seizures accelerate anoxia-induced neuronal death in the neonatal rat hippocampus. AB - Seizures occurring in infants with hypoxia are frequently associated with an ominous prognosis. There is, however, no direct evidence that seizures are involved in the pathogenesis of hypoxia-induced neuronal damage. Here, we report that seizures significantly aggravate the hypoxic state by accelerating rapid anoxic depolarization (AD) and associated neuronal death in preparations of the intact hippocampus of neonatal rats in vitro. Under control conditions, prolonged episodes of anoxia/aglycemia induced rapid suppression of synaptic activity followed sequentially by brief bursts of epileptiform activity and then by rapid AD. AD was associated with irreversible neuronal damage manifested by irreversible loss of the membrane potential, synaptic responses, and neuronal degeneration. Aggravation of electrographic seizure activity during anoxic episodes by the adenosine A1 receptor antagonists DPCPX and caffeine or the gamma aminobutyric acid-A receptor antagonist bicuculline or pretreatment with 4 aminopyridine accelerated AD and associated neuronal death by up to twofold, whereas blockade of seizure activity by the glutamate receptor antagonists or tetrodotoxin significantly delayed the onset of AD. This report provides direct evidence for the need to prevent seizures during neonatal brain hypoxia. PMID- 11026448 TI - Innate production of interleukin-10 and tumor necrosis factor affects the risk of multiple sclerosis. AB - Multiple sclerosis (MS) typically presents with a relapsing-remitting onset. This can be distinguished from primary progressive MS. Typical MS is characterized by a profound inflammatory reaction in which anti-inflammatory cytokine interleukin 10 (IL-10) and pro-inflammatory cytokine tumor necrosis factor (TNF) may play a pivotal role. We tested the hypothesis that patients with MS have a distinct innate cytokine production that contributes to the susceptibility for and outcome of MS. The innate cytokine production of patients was estimated as the average production of cytokines in lipopolysaccharide -stimulated whole-blood cultures of 2 to 5 first-degree healthy family members. A total of 126 family members of 50 patients with typical MS, 61 family members of 25 patients with primary progressive MS, and 129 control subjects of 54 families were enrolled in this study. We found that members of families with low IL-10 and high TNF production had a fourfold increased risk of developing typical MS compared with members of families with high IL-10 and low TNF production. Patients with MS were eightfold more likely to develop typical MS than primary progressive MS when they belonged to families with low IL-10 and high TNF production. The presence of human leukocyte antigen-DR2 was associated with MS but not with TNF production. This study shows that typical MS is associated with an innate pro-inflammatory cytokine profile in contrast to primary progressive MS. PMID- 11026449 TI - Clinical, genetic, and expression studies of mutations in the potassium channel gene KCNA1 reveal new phenotypic variability. AB - Episodic ataxia type 1 (EA1) is an autosomal dominant central nervous system potassium channelopathy characterized by brief attacks of cerebellar ataxia and continuous interictal myokymia. Point mutations in the voltage-gated potassium channel gene KCNA1 on chromosome 12p associate with EA1. We have studied 4 families and identified three new and one previously reported heterozygous point mutations in this gene. Affected members in Family A (KCNA1 G724C) exhibit partial epilepsy and myokymia but no ataxic episodes, supporting the suggestion that there is an association between mutations of KCNA1 and epilepsy. Affected members in Family B (KCNA1 C731A) exhibit myokymia alone, suggesting a new phenotype of isolated myokymia. Family C harbors the first truncation to be reported in KCNA1 (C1249T) and exhibits remarkably drug-resistant EA1. Affected members in Family D (KCNA1 G1210A) exhibit attacks typical of EA1. This mutation has recently been reported in an apparently unrelated family, although no functional studies were attempted. Heterologous expression of the proteins encoded by the mutant KCNA1 genes suggest that the four point mutations impair delayed-rectifier type potassium currents by different mechanisms. Increased neuronal excitability is likely to be the common pathophysiological basis for the disease in these families. The degree and nature of the potassium channel dysfunction may be relevant to the new phenotypic observations reported in this study. PMID- 11026450 TI - Cortical areas differentially involved in multiplication and subtraction: a functional magnetic resonance imaging study and correlation with a case of selective acalculia. AB - A patient with an intracranial hemorrhage showed differential impairment among arithmetic types (impaired in multiplication but not in subtraction). A functional magnetic resonance imaging (fMRI) experiment using normal volunteers also revealed a differential activation between the two arithmetic types. The fMRI result could account for the selective acalculia of the patient in that the lesion shown by structural MRI included the region with multiplication-higher activation and spared subtraction-higher regions. These findings suggest that the cognitive mechanism and neural substrates differ among the simple arithmetic operations. PMID- 11026451 TI - Neurolisteriosis presenting as recurrent transient ischemic attacks. AB - An elderly man experienced recurrent transient episodes of right arm weakness and expressive aphasia. He was initially treated with aspirin and then with coumadin. Thirteen days after initial presentation, he became febrile and had signs of meningitis. The illness progressed relentlessly to death 9 weeks after admission to the hospital. Necropsy showed prominent meningitis with vasculitis extending into the left frontal lobe. Polymerase chain reaction identified the organism as Listeria monocytogenes. PMID- 11026452 TI - Sporadic fatal insomnia: a case study. AB - A 58-year-old man died after a 27-month illness characterized by insomnia, confirmed by polysomnography. He was homozygous for methionine at codon 129 of the prion gene but had no mutation in the prion gene. Neuropathology showed thalamic and olivary atrophy and no spongiform changes. Paraffin-embedded tissue blotting demonstrated abnormal prion protein in the brain. This is the first case of the sporadic form of fatal familial insomnia with demonstration of the disorder by polysomnography. PMID- 11026453 TI - Aneurysm of a dural sigmoid sinus: a novel vascular cause of pulsatile tinnitus. AB - We report a newly evidenced cause of venous pulsatile tinnitus--the aneurysm of a dural sigmoid sinus. A 33-year-old patient presented with an incapacitating pulsatile tinnitus of 6 months' duration in the left ear. The radiological workup evidenced an aneurysm of the left sigmoid sinus. Selective endovascular coil occlusion of the aneurysm was followed by complete resolution of the tinnitus. PMID- 11026454 TI - Peripheral neuropathy with hypomyelination, chronic intestinal pseudo-obstruction and deafness: a developmental "neural crest syndrome" related to a SOX10 mutation. AB - We describe the case of a girl with an unusual congenital phenotype, combining peculiar peripheral nerve lesions with hypomyelination, chronic intestinal pseudoobstruction, and deafness. She was found to have a de novo heterozygous frameshift mutation in the gene encoding the SOX10 transcription factor. The likely role of SOX10 in determining the fate of Schwann cells during early embryogenesis may explain the peripheral nervous system developmental disorder observed in this patient. PMID- 11026455 TI - Impaired activation of oxygen consumption and blood flow in visual cortex of patients with mitochondrial encephalomyopathy. AB - A current hypothesis claims that an increase of blood flow is required for oxygen consumption to rise during neuronal excitation (activation). Chronic progressive external ophthalmoplegia is a mitochondrial disease associated with deletions of mtDNA or by point mutation of tRNA genes. We tested the hypothesis that the cerebral metabolic rate of oxygen (CMRO2) may not rise in this disorder if the accompanying cerebral blood flow increase is insufficient. Two patients with progressive external ophthalmoplegia were visually stimulated with a colored checkerboard pattern reversing as different frequencies. When stimulated, Patient 1 had a small increase of cerebral blood flow, while Patient 2 had no cerebral blood flow increase. In the visually active state, the patients had no significant change of CMRO2, while healthy subjects had a pronounced increase of CMRO2 in the pericalcarine visual cortex at 4 Hz and a further slight increase at 8 Hz during activation. PMID- 11026456 TI - Muscle fibers in inflammatory myopathies and cultured myoblasts express the nonclassical major histocompatibility antigen HLA-G. AB - We demonstrate that HLA-G, a nonclassical major histocompatibility complex class I antigen, is expressed in muscle fibers in various inflammatory myopathies. Further, interferon-gamma induces surface expression and upregulation of mRNA transcripts corresponding to different isoforms of HLA-G in myoblasts cultured from control subjects and patients. HLA-G may have important immunological functions in inflammatory myopathies and other local immune reactions as they occur during vaccination, myoblast transplantation, and gene therapy. PMID- 11026457 TI - NOS3 polymorphism not associated with Alzheimer's disease in Japanese. PMID- 11026459 TI - NCI initiative: development of novel imaging technologies. PMID- 11026458 TI - Lyophilized red wine administration prolongs survival in an animal model of amyotrophic lateral sclerosis. PMID- 11026460 TI - Gray-scale skeletonization of small vessels in magnetic resonance angiography. AB - Interpretation of magnetic resonance angiography (MRA) is problematic due to complexities of vascular shape and to artifacts such as the partial volume effect. We present new methods to assist in the interpretation of MRA. These include methods for detection of vessel paths and for determination of branching patterns of vascular trees. They are based on the ordered region growing (ORG) algorithm that represents the image as an acyclic graph, which can be reduced to a skeleton by specifying vessel endpoints or by a pruning process. Ambiguities in the vessel branching due to vessel overlap are effectively resolved by heuristic methods that incorporate a priori knowledge of bifurcation spacing. Vessel paths are detected at interactive speeds on a 500-MHz processor using vessel endpoints. These methods apply best to smaller vessels where the image intensity peaks at the center of the lumen which, for the abdominal MRA, includes vessels whose diameter is less than 1 cm. PMID- 11026461 TI - Using an MRI distortion transfer function to characterize the ghosts in motion corrupted images. AB - Motion artefact suppression remains an active topic in MRI. In this paper, we suggest that certain nonrigid, or spatially variant, characteristics of motion of an object can be represented by extending the work of Mitsa et al. This empirical extension uses a ghost distortion transfer function (GTDF) applied to the k-space (frequency domain) data. We demonstrate the variety of ghost characteristics that can be generated from various two-dimensional (2-D) GTDF's. The distortion transfer function for periodic motion along the Z-axis can be determined from the nonoverlapped portions of the ghost and central image. It required a GDTF with the shape of a belt bandpass filter to produce an image corresponding to the ghosts of a volunteer's abdomen image corrupted by unknown respiratory motion artefacts. The preliminary results of a composite method of motion artefact suppression are presented. The artefact suppression was successful for ghost images described by a GDTF have a low-pass nature, but less successful with ghosts have a GDTF of a bandpass nature. PMID- 11026462 TI - A filter design method for minimizing ringing in a region of interest in MR spectroscopic images. AB - Magnetic resonance spectroscopic imaging (MRSI) requires a relatively long time to sample k-space (the spatial frequency domain), effectively lowpass filtering the resulting reconstructed image. Ringing is especially problematic when a region of interest (ROI) is close to a bright region outside the ROI, since the bright region tends to create a ringing artifact into the ROI due to the lowpass nature of the data. In this paper, we propose a method that reduces the effect of a stronger signal region on a weaker signal in a nearby ROI by designing a postprocessing filter that steers the strong interference away from the ROI. The proposed method is computationally simple both in the design stage and in applying it to images. We present experiments that illustrate the value of the technique. PMID- 11026463 TI - Regularization for uniform spatial resolution properties in penalized-likelihood image reconstruction. AB - Traditional space-invariant regularization methods in tomographic image reconstruction using penalized-likelihood estimators produce images with nonuniform spatial resolution properties. The local point spread functions that quantify the smoothing properties of such estimators are space-variant, asymmetric, and object-dependent even for space-invariant imaging systems. We propose a new quadratic regularization scheme for tomographic imaging systems that yields increased spatial uniformity and is motivated by the least-squares fitting of a parameterized local impulse response to a desired global response. We have developed computationally efficient methods for PET systems with shift invariant geometric responses. We demonstrate the increased spatial uniformity of this new method versus conventional quadratic regularization schemes in simulated PET thorax scans. PMID- 11026464 TI - A modular software system for computer-aided surgery and its first application in oral implantology. AB - Development of complex software applications in image guided therapy (IGT) is often complicated by the fact that providing basic functionality for image processing and user interaction from a graphical user interfaces (GUI) requires considerable manpower for software development. We present a programming environment that combines the high-level image processing library AVW, in-house developed patient-to-image registration procedures, and an interface to position measurement hardware. A specific application can be developed by using Tcl/Tk, a simple platform-independent scripting language, for GUI development. This environment was applied to developing VISIT, a navigation system for computer aided implant dentistry. VISIT is presented as a result of this paper. Parts of the development environment were made accessible to the public. Future work includes the implementation of video input for connecting ultrasound or fluoroscopy units. We conclude that our approach is well suited to accelerate the development of exploratory new applications of IGT. PMID- 11026465 TI - Ultrasound measurement of brachial flow-mediated vasodilator response. AB - Brachial artery flow-mediated vasodilation is increasingly used as a measure of endothelial function. High resolution ultrasound provides a noninvasive method to observe this flow-mediated vasodilation by monitoring the diameter of the artery over time following a transient flow stimulus. Since hundreds of ultrasound images are required to continuously monitor brachial diameter for the 2-3 min during which the vasodilator response occurs, an automated diameter estimation is desirable. However, vascular ultrasound images suffer from structural noise caused by the constructive and destructive interference of the backscattered signals, and the true boundaries of interest that define the diameter are frequently obscured by the multiple-layer structure of the vessel wall. These problems make automated diameter estimation strategies based on the detection of the vessel wall boundary difficult. We obtain a robust automated measurement of the vasodilator response by automatically locating the artery using a variable window method, which gives both the lumen center and width. The vessel wall boundary is detected by a global constraint deformable model, which is insensitive to the structural noise in the boundary area. The ambiguity between the desired boundary and other undesired boundaries is resolved by a spatiotemporal strategy. Our method provides excellent reproducibility both for interreader and intrareader analyzes of percent change in diameter, and has been successfully used in analyzing over 4000 brachial flow-mediated vasodilation scans from several medical centers in the United States. PMID- 11026466 TI - Analysis of geometrical distortion and statistical variance in length, area, and volume in a linearly scanned 3-D ultrasound image. AB - A linearly scanned three-dimensional (3-D) ultrasound imaging system is considered. The transducer array is initially oriented along the x axis and aimed in the y direction. After being tilted by an angle theta about the x axis, and then swiveled by an angle phi about the y axis, it is translated in the z direction, in steps of size d, to acquire a series of parallel two-dimendional (2 D) images. From these, the 3-D image is reconstructed, using the nominal values of the parameters (phi, theta, d). Thus, any systematic or random errors in these, relative to their actual values (phi0, theta0, d0), will respectively cause distortions or variances in length, area, and volume in the reconstructed 3 D image, relative to the 3-D object. Here, we analyze these effects. Compact linear approximations are derived for the relative distortions as functions of the parameter errors, and hence, for the relative variances as functions of the parameter variances. Also, exact matrix formulas for the relative distortions are derived for arbitrary values of (phi, theta, d) and (phi0, theta0, d0). These were numerically compared to the linear approximations and to measurements from simulated 3-D images of a cubical object and real 3-D images of a wire phantom. In every case tested, the theory was confirmed within experimental error (0.5%). PMID- 11026467 TI - Deformable boundary detection of stents in angiographic images. AB - In this paper, a procedure is described for deformable boundary detection of medical tools, called stents, in angiographic images. A stent is a surgical stainless steel coil that is placed in the artery in order to improve blood circulation in regions where a stenosis has appeared. Assuming initially a set of three-dimensional (3-D) models of stents and using perspective projection of various deformations of the 3-D model of the stent, a large set of synthetic two dimensional (2-D) images of stents is constructed. These synthetic images are then used as a training set for deriving a multivariate Gaussian density estimate based on eigenspace decomposition and formulating a maximum-likelihood estimation framework in order to reach an initial rough estimate for automatic object recognition. The silhouette of the detected stent is then refined by using a 2-D active contour (snake) algorithm integrated with a novel iterative initialization technique, which takes into consideration the geometry of the stent. The algorithm is experimentally evaluated using real angiographic images containing stents. PMID- 11026468 TI - Excitatory amino acids studied in isolated cells and culture preparations: a bridge over in vitro and in vivo experimental paradigms. PMID- 11026469 TI - Excitatory amino acid neurotoxicity and modulation of glutamate receptor expression in organotypic brain slice cultures. AB - Using organotypic slice cultures of hippocampus and cortex-striatum from newborn to 7 day old rats, we are currently studying the excitotoxic effects of kainic acid (KA), AMPA and NMDA and the neuroprotective effects of glutamate receptor blockers, like NBQX. For detection and quantitation of the induced neurodegeneration, we have developed standardized protocols, including--a) densitometric measurements of the cellular uptake of propidium iodide (PI), --b) histological staining by Flouro-Jade, --c) lactate dehydrogenase (LDH) release to the culture medium, --d) immunostaining for microtubulin-associated protein 2, and --e) general and specific neuronal and glial cell stains. The results show good correlation between the different markers, and are in accordance with results obtained in vivo. Examples presented in this review will focus on the use of PI uptake to monitor the excitotoxic effects of --a) KA and AMPA (and NMDA) in hippocampal slice cultures, and --b) KA and AMPA in corticostriatal slice cocultures, with demonstration of differentiated neuroprotective effects of NBQX in relation to cortex and striatum and KA and AMPA. A second set of studies include modulation of hippocampal KA-induced excitotoxicity and KA-glutamate receptor subunit mRNA expression after long-term exposure to low, non-toxic doses of KA and NBQX. We conclude that organotypic brain slice cultures, combined with standardized procedures for quantitation of cell damage and receptor subunit changes is of great potential use for studies of excitotoxic, glutamate receptor induced neuronal cell death, receptor modulation and related neuroprotection. PMID- 11026470 TI - Margatoxin and iberiotoxin, two selective potassium channel inhibitors, induce c fos like protein and mRNA in rat organotypic dorsal striatal slices. AB - The isolated single organotypic slice model allows to investigate the effects of drugs and toxins on the expression of transcription factors in the striatum without dopaminergic and glutamatergic interactions. In this study the effects of margatoxin and iberiotoxin on the expression of c-fos mRNA by in situ hybridization as well as on c-fos like protein by immunohistochemistry in isolated dorsal striatum after 10 days in culture were investigated. C-fos mRNA dose-dependently increased 30 min after incubation with margatoxin and iberiotoxin. Expression of c-fos like protein was transiently detected 3h afterwards. This effect is independent from extrinsic neuronal circuitry as dopamine neurons were found to be absent in the cultured slices. It is concluded that inhibition of voltage-gated as well as calcium-activated (Slo) potassium channels leads to activation of gene transcription in striatal neurons which may trigger long-term changes in transmitter plasticity. PMID- 11026471 TI - Excitatory amino acids, monoamine, and nitric oxide synthase systems in organotypic cultures: biochemical and immunohistochemical analysis. AB - The nigrostriatal and mesolimbic systems of the rat have been re-constructed using the organotypic culture model, whereby neonatal brain tissue is grown in vitro for approximately one month. The nigrostriatal cultures consisted of tissue from the substantia nigra, dorsal striatum and frontoparietal cortex; while the mesolimbic cultures included the ventral tegmental area, ventral striatum and cingulate cortex. The cultures were grown at 35 degrees C in normal atmosphere, using a tuberoller device placed in a cell incubator and changing the medium every 3-4 days. The in vitro development was evaluated with an inverted microscope equipped with a variable relief contrast function. Samples were taken directly from the medium in the culture tube and analysed for several amino acids with HPLC. After a month the cultures were fixed and processed for immunohistochemistry. High levels of glutamate and aspartate were observed every time the medium was changed, but the levels rapidly decreased reaching a steady state after approximately 24h. A decrease in the levels was also observed along development, reaching stable values (approximately 2 microM and approximately 0.12 microM for glutamate and aspartate, respectively) at approximately two weeks, but only when the cultures showed an apparently healthy development. The levels were approximately 10 times higher in deteriorating or apparently damaged cultures. Glutamine levels were in the mM range and remained stable along the entire experiment. No differences were observed among nigrostriatal and mesolimbic cultures. Immunohistochemistry confirmed the impressions obtained from microscopic and biochemical analysis along the in vitro development, revealing apparently healthy neuronal systems with characteristics similar to those observed in vivo, when tyrosine hydroxylase and nitric oxide synthase, markers for dopamine and nitric oxide containing neurons, respectively, were analysed. In the substantia nigra, nitric oxide synthase-positive networks surrounded tyrosine hydroxylase-positive neurons, while in the striatum nitric oxide synthase dendrites were surrounded by tyrosine hydroxylase-positive nerve terminals, suggesting a reciprocal interaction among dopamine and nitric oxide containing neurons. Thus, the organotypic model appears to capture many of the neurochemical and morphological features seen in vivo, providing a valuable model for studying in detail the neurocircuitries of the brain. PMID- 11026472 TI - Dale's principle and glutamate corelease from ventral midbrain dopamine neurons. AB - While direct application of dopamine modulates postsynaptic activity, electrical stimulation of dopamine neurons typically evokes excitation. Most of this excitation appears to be due to activation of collateral pathways; however, several lines of evidence have suggested that there is a monosynaptic component due to glutamate corelease by dopamine neurons. Recently, more direct evidence obtained in culture has shown that ventral midbrain dopamine neurons release both dopamine and glutamate. Moreover, they appear to do so from separate release sites, calling into question recent modifications of Dale's Principle. The neurochemical phenotype of a given synapse may be determined by subcellular neurotransmitter levels, uptake, or storage. However, the relationship between dopamine and glutamate release from dopamine neuron synapses in the intact brain- and the mechanisms involved--has yet to be resolved. PMID- 11026473 TI - Diversity of glutamate transporter expression and function in the mammalian retina. AB - Glutamate is the major excitatory neurotransmitter of the mammalian retina and glutamate uptake is essential for normal transmission at glutamatergic synapses. Between photoreceptors and second order neurons, increases in light intensity are signaled by decreases in the concentration of glutamate within the synaptic cleft. In such a system the precise control of glutamate in the synaptic cleft is thus essential and glutamate transporters are thought to contribute to this process. As demonstrated here, all neuronal and macroglial cells of the retina appear to express high-affinity glutamate transporters. GLAST1, GLT1, EAAC1 and EAAT5 are expressed in the retina and exhibit unique localisation and functional properties. In the present study we summarize retinal glutamate transporter expression, identify the major glutamate uptake site in the mammalian retina and discuss the possible functional roles of different glutamate transporter subtypes in glutamatergic neurotranmission in the retina. PMID- 11026474 TI - Lack of coupling between GABA release and GABA synthesis in the rat brain via GABA(B) autoreceptors. AB - GABA is synthesized within GABA terminals through a highly compartmentalized process in which glial-derived glutamine is a major precursor and its release is modulated by GABA(B) autoreceptors. The aim of this work was to ascertain whether or not GABA synthesis and release are coupled in the rat brain through a GABA(B) autoreceptor-mediated modulation. It was found that (-)baclofen (30microM) reduces the K+ stimulated release of [3H]GABA in synaptosomes and prisms (10microM) from cerebral cortex, while at the same concentrations (-)baclofen failed to modify the synthesis of [3H]GABA from [3H]glutamine in cortical and hypothalamic slices, prisms and in cortical synaptosomes. In this latter preparation, identical results were observed when (-)baclofen was added to Krebs Tris media, containing 5 or 15 mM K+ concentration. In agreement with these latter results, glutamic acid decarboxylase (GAD) activity from cortical and hypothalamic prisms was not affected by 1-100 microM (-)baclofen. Similar results on GABA synthesis were also observed when 1-100 microM 3 aminopropil(methyl)phosphinic acid or GABA was used instead of (-)baclofen to stimulate GABA(B) autoreceptors. [3H]GABA release, [3H]GABA synthesis from [3H]glutamine and GAD activity were also insensitive to the action of the GABA(B) antagonist CGP 52432 (10-100microM). Likewise, muscimol (0.3-100microM) did not affect GABA synthesis. Our results indicate that unlike GABA release, GABA synthesis is not modulated by GABA(B) autoreceptors. PMID- 11026475 TI - Metabotropic glutamate receptors--different therapeutic perspectives. PMID- 11026476 TI - Stimulation of group II metabotropic glutamate receptors or inhibition of group I ones exerts anxiolytic-like effects in rats. AB - Using the conflict drinking Vogel test in rats as a model we examined the anxiolytic-like activity of (S)-4-carboxyphenylglycine (S-4CPG), an antagonist of group I metabotropic glutamate receptors (mGlu receptors), of (RS)-a-methylserine O-phosphate-monophenyl ester (MSOPPE), an antagonist of group II mGlu receptors, and of (2S,1'S,2'S)-2-(carboxycyclopropyl)glycine (L-CCG-I), an agonist of group II mGlu receptors. The obtained results indicate that intrahippocampal administration of S-4CPG and L-CCG-I, but not MSOPPE to rats produces a dose dependent anticonflict effect, which is unrelated to the reduced perception of the stimulus or to an increased thirst drive. The hippocampus may be one of the neuroanatomical sites of the anxiolytic-like effects of either agent. PMID- 11026477 TI - The role of glutamate receptors in antipsychotic drug action. AB - It has recently been postulated that disturbances in glutamatergic neurotransmission may contribute to the pathophysiology of schizophrenia. Therefore the aim of the present study was to evaluate the role of glutamate NMDA and group II metabotropic receptors in the antipsychotic drug action. To this aim the influence of some well-known neuroleptics on cortical NMDA receptors was examined. Furthermore, their behavioral effects were compared with those of the novel agonist of group II glutamate metabotropic receptors, LY 354740, in some animal models of schizophrenic deficits. We found that long-term administration of the typical neuroleptic haloperidol and the atypical one clozapine increased the number of NMDA receptors labelled with [3H]CGP 39653 in different cortical areas. Long-, but not short-term, treatment with haloperidol and raclopride diminished the deficit of prepulse inhibition produced by phencyclidine, which is a model of sensorimotor gating deficit in schizophrenia. In contrast, neither short- nor long-term treatment with clozapine influenced the phencyclidine effect in that model. Acute treatment with LY 354740 reversed neither (1) the deficit of prepulse inhibition produced by phencyclidine or apomorphine, nor (2) the impairment in a delayed alternation task induced by MK-801, which is commonly used to model the frontal lobe deficits associated with schizophrenia. The present study suggests that an increase in the density of cortical NMDA receptors may be important to a longterm neuroleptic therapy. Conversely, the results do not support the role of group II metabotropic glutamate receptors in the antipsychotic drug action. PMID- 11026478 TI - The role of metabotropic glutamate receptor (mGluR) ligands in parkinsonian muscle rigidity. AB - It has been shown that the primary striatal dopaminergic hypofunction which is at the origin of Parkinson's disease, results in a secondary hyperactivity of glutamatergic neurotransmission. In the search for a therapy of Parkinson's disease, ionotropic, mainly NMDA, receptor antagonists were found to have moderately beneficial, yet also some undesirable side-effects. Therefore the present study was aimed at determining whether some metabotropic glutamate receptor (mGluR) ligands may have antiparkinsonian effects in the haloperidol induced muscle rigidity. To this end three mGluR ligands were used: the potent and selective mGluR I antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA), the mixed group II agonist/ group I antagonist (S)-4-carboxy-3-hydroxyphenyl glycine ((S)-4-C3HPG), and the potent group II agonist (+)-2 aminobicyclo[3.1.0.]hexane-2,6,-dicarboxylic acid (LY354740). Only LY354740 penetrated the brain from the periphery; for this reason other drugs were injected bilaterally into the rostral striatum or nucleus accumbens. The muscle tone was recorded by a mechanomyographic/electromyographic (MMG/EMG) method which measured the resistance of a rat's hind foot and the EMG reflex response of its muscles to passive movements. (S)-4C3HPG (5 and 15 microg/0.5 microl) and LY354740 (5 and 10mg/kg i.p.) diminished the muscle rigidity induced by haloperidol (1 mg/kg i.p.). AIDA (0.5-15 microg/0.5 microl) injected into the striatum was only slightly effective in the highest dose used. However, when injected into the nucleus accumbens AIDA (15microg/0.5microl) significantly and strongly counteracted the haloperidol-induced muscle rigidity. Our results suggest that stimulation of group II striatal mGluRs seems to play a major role in diminution of parkinsonian-like muscle rigidity. However, it seems that the antagonism of group I mGluRs located in the nucleus accumbens may also be of importance to the antiparkinsonian effect. PMID- 11026479 TI - On the role of inhibitory glutamate receptors in N-methyl-D-aspartate- and dopamine-receptor mediated motor behavior of rats. AB - The physiological function of inhibitory group II metabotropic glutamate receptors, a family of second messenger coupled glutamate-receptors, for motor behavior is almost unknown. The aim of this study is to address this topic by quantifying motor effects of the preferential group II agonist (2S,3S,4S)-alpha (carboxycyclopropyl)-glycine, administered i.c.v. (62.5, 125.0, 187.5, 250.0, 500.0nmol/4microl), in an open-field equipped with a hole-board. (2S,3S,4S)-alpha (carboxycyclopropyl)-glycine decreased spontaneous locomotor and exploratory behavior, which was blocked be the group II antagonist (2S)-alpha-ethylglutamic acid (250.0nmol/4microl). Locomotion induced by the N-methyl-D-aspartate-receptor antagonist dizocilpine (0.08, 0.16, 0.32mg/kg) was counteracted by the group II agonist (2S,3S,4S)-alpha-(carboxycyclopropyl)-glycine, but an antagonism towards dizocilpine did not occur in all aspects of motor behavior evaluated. In contrast to the antagonism of dizocilpine induced locomotion, D,L-amphetamine (1.0, 2.0, 3.0mg/kg) induced locomotion was not antagonised by (2S,3S,4S)-alpha (carboxycyclopropyl)-glycine.The results suggest that group II agonists may be devoid of psychotomimetic effects in humans and even may antagonise this side effect of N-methyl-D-aspartate receptor antagonists. Since group II activation and N-methyl-D-aspartate-receptor blockade very efficiently protects against excitotoxic neurodegeneration, selective group II agonists may allow novel pharmacotherapeutical approaches in pathophysiological conditions characterised by a glutamatergic hyperactivity, like epilepsy, ischemia and trauma. PMID- 11026480 TI - Cell-type specificity of mGluR activation in striatal neuronal subtypes. AB - The effects of metabotropic glutamate receptor (mGluR) activation were studied in medium spiny neurons and large aspiny (LA) interneurons by means of electrophysiological and optical recordings. DCG-IV and L-SOP, agonists for group II and III mGluRs, respectively, produced a presynaptic inhibitory effect on corticostriatal glutamatergic excitatory postsynaptic potentials (EPSPs) in both spiny and LA cells. Activation of group I mGluRs by the selective agonist 3,5 DHPG produced no effect on membrane properties and glutamatergic transmission in spiny neurons, whereas it did cause a membrane depolarization in LA interneurons coupled to increased input resistance. In combined optical and electrophysiological experiments, in spiny neurons 3,5-DHPG enhanced membrane depolarization and intracellular calcium (Ca2+) levels induced by NMDA applications, but not in LA interneurons. These data suggest the existence of a positive interaction between NMDA and group I mGlu receptors only in medium spiny cells which might, at least partially, account for the differential vulnerability to excitotoxic damage observed in striatal neuronal subtypes. PMID- 11026481 TI - Uncompetitive NMDA receptor antagonists--is low affinity better? PMID- 11026482 TI - Low affinity channel blocking (uncompetitive) NMDA receptor antagonists as therapeutic agents--toward an understanding of their favorable tolerability. AB - Studies in experimental models have suggested that NMDA receptor antagonists may have utility in the treatment of a wide variety of neurological and psychiatric disorders. However, clinical trials have not been encouraging largely because the antagonists evaluated to date have exhibited unacceptable neurobehavioral side effects. In animals, therapeutic doses of some low-affinity channel blocking (uncompetitive) NMDA receptor antagonists are associated with less gross neurological impairment and behavioral toxicity than other types of NMDA receptor antagonists. Favorable clinical experiences with several such agents has bolstered confidence in the neurotherapeutic potential of low affinity NMDA antagonists. This article reviews current research attempting to explain the improved tolerability of such antagonists. While no single mechanism appears to account for the reduced toxicity of such agents, kinetic properties, particularly rapid blocking rate, seem to be of key importance. Other factors include partial trapping, reduced agonist-independent (closed channel) block, subunit selectivity (particularly for receptors that do not contain the NR2A subunit), combined block at allosteric (voltage-independent) sites, and synergistic therapeutic effects produced by additional actions at receptor targets apart from NMDA receptors (e.g., weak positive allosteric modulation of GABA(A) receptors or state dependent Na+ channel block). PMID- 11026483 TI - Low affinity use-dependent NMDA receptor antagonists show promise for clinical development. AB - The success of the low affinity use-dependent NMDA receptor antagonists to reach clinical trials can be readily attributed to their wider margins of safety and lack of neurotoxicity at higher doses. Several mechanistic differences distinguish the low affinity from the high affinity use-dependent antagonists: 1) Differential regional affinities for the various NMDA receptor subtypes; 2) The static receptor blockade due to the faster on/off rate receptor kinetics which limit, but do not totally prevent the amount of Ca+2 entry into the cell during glutamate-induced depolarization; and 3) Rapid egress of the compounds from the ion channel during recovery resulting in less membrane trapping between transmission pulses. Advanced clinical trials are in progress for the following indications: epilepsy, stroke, head trauma, tardive dyskinesia, pain plus Parkinson's, Huntington's and Alzheimer's diseases. PMID- 11026484 TI - Memantine and the amino-alkyl-cyclohexane MRZ 2/579 are moderate affinity uncompetitive NMDA receptor antagonists--in vitro characterisation. AB - There is general agreement that moderate affinity uncompetitive NMDA receptor antagonists combine good efficacy and tolerability in animal models of disturbances in glutamatergic transmission. There are several theories on which properties are important for this profile including 1, rapid access to the channel at the start of pathological overactivity 2, rapid, voltage-dependent relief of blockade during physiological synaptic activation and 3, partial untrapping. Merz has developed a series of novel uncompetitive NMDA receptor antagonists based on the cyclohexane structure. In cultured hippocampal neurones MRZ 2/579 (1-amino-1,3,3,5,5-pentamethylcyclohexane) shows similar blocking kinetics to memantine (Kon 10.7 * 10(4)M(-1)sec(-1), Koff 0.20sec(-1) at -70mV) and binds at the same depth in the NMDA receptor channel (delta = 0.8). The potency of MRZ 2/579 assessed as Kd = Koff/Kon = 1.87microM agrees well with the IC50 of 1.29microM against steady-state currents in cultured hippocampal neurones (at -70mV) and with the Ki in [3H]-MK-801 binding of 0.65microM. MRZ 2/579 protected cultured cortical neurones against glutamate toxicity with an IC50 of 2.16microM and was also effective in protecting hippocampal slices against hypoxia/hypoglycaemia-induced reduction of fEPSP amplitude in CA1 with an EC50 of 7.01microM. MRZ 2/579 has similar potency and bio-availability to memantine in vivo assessed using microdialysis, microiontophoresis and MES-induced seizures. Initial characterization in animal models provides strong support for the assumption that MRZ 2/579 could be a useful therapeutic in morphine/alcohol dependence, inhibition of morphine tolerance, chronic pain and as a neuroprotective agent. PMID- 11026485 TI - NMDA channel blockers: memantine and amino-aklylcyclohexanes--in vivo characterization. AB - The previous overviews provided the basis for better therapeutic efficacy/tolerability of low to moderate affinity NMDA channel blockers. This prediction finds support in in vitro studies comparing protective and plasticity impairing effects (therapeutic vs. side-effect) of memantine and (+)MK-801. In fact it turned out that memantine had a far better therapeutic index. But can it be confirmed in the in vivo situation? PMID- 11026486 TI - Differences in degree of trapping between AR-R15896 and other uncompetitive NMDA receptor antagonists. AB - NMDA antagonists like AR-R15896 have been selected on the basis of their good therapeutic indices. As Dr. Rogawski has pointed out, there may be a number of molecular factors which can improve the therapeutic index of NMDA antagonists. In this paper we will consider three factors; use-dependence, low affinity/fast kinetics, and partial trapping. PMID- 11026487 TI - NPS 1506, a moderate affinity uncompetitive NMDA receptor antagonist: preclinical summary and clinical experience. AB - NPS Pharmaceuticals, Inc. (NPS) has synthesized a series of open-channel blockers with varying potencies at the NMDA receptor. NPS 1506 (Fig. 1) is a moderate affinity antagonist that inhibits NMDA/glycine-induced increases in cytosolic calcium in cultured rat cerebellar granule cells (IC50 = 476nM) and displaces the binding of [3H]MK-801 to rat cortical membranes (IC50 = 664nM). PMID- 11026488 TI - Neurobiology of the ventral basal ganglia loop. PMID- 11026489 TI - Dopaminergic denervation enhances susceptibility to hydroxyl radicals in rat neostriatum. AB - To determine if greater amounts of hydroxyl radical (*OH) are formed by dopamine (DA) denervation and treatment with L-dihydroxyphenylalanine (L-DOPA), the neostriatum was DA denervated (99% reduction in DA content) by 6-hydroxydopamine treatment (134microg icv, desipramine pretreatment) of neonatal rats. At 10 weeks the peripherally restricted dopa decarboxylase inhibitor carbidopa (12.5mg/kg i.p.) was administered 30min before vehicle, L-DOPA (60mg/kg i.p.), or the known generator of reactive oxygen species, 6-hydroxydopa (6-OHDOPA) (60mg/kg i.p.); and this was followed 30min later (and 15 min before termination) by the spin trap, salicylic acid (8micromoles icv). By means of a high performance liquid chromatographic method with electrochemical detection, we found a 4-fold increase in the non-enzymatically formed spin trap product, 2,3-dihydroxybenzoic acid (2,3 DHBA), with neither L-DOPA nor 6-OHDOPA having an effect on 2,3-DHBA content of the neostriatum. Basal content of 2,5-DHBA, the enzymatically formed spin trap product, was 4-fold higher vs. 2,3-DHBA in the neostriatum of untreated rats, while L-DOPA and 6-OHDOPA each reduced formation of 2,5-DHBA. We conclude that DA innervation normally suppresses *OH formation, and that the antiparkinsonian drug L-DOPA has no effect (2,3-DHBA) or slightly reduces (2,5-DHBA) *OH formation in the neostriatum, probably by virtue of its bathing the system of newly formed *OH. PMID- 11026490 TI - Functional aspects of the ventral pallidum. AB - The ventral pallidum is part of the corticoaccumbo-thalamocortical loop of the basal ganglia. In the past the function of this structure was discussed as a pure relay station in the process of limbic-motor integration. Some recent studies, however, underline that on the level of the ventral pallidum motor behavior can be modulated. The stimulation and inhibition of the different transmitter systems that converge in the ventral pallidum (dopamine, glutamate, GABA, neuropeptides) have implications in repetitive-, disinhibited-, learning- and reinforced behavior. The present review summarizes available data of these parameter related to this behavior, i.e. locomotion, reward-related behavior, prepulse inhibition, memory and neurochemistry. PMID- 11026491 TI - The medial prefrontal cortex as a part of the brain reward system. AB - This review will briefly summarize experimental evidence for an involvement of the medial prefrontal cortex (mPFC) in reward-related mechanisms in the rat brain. The mPFC is part of the mesocorticolimbic dopaminergic system. It receives prominent dopaminergic input from the ventral tegmental area (VTA) and, via the mediodorsal thalamus, inputs from other subcortical basal ganglia structures. In turn it projects back to the VTA and the nucleus accumbens septi (NAS), which are generally considered as main components of the brain reward system. Evidence for the involvement of the mPFC in reward-related mechanisms comes mainly from three types of studies, conditioned place preference (CPP), intracranial self stimulation (ICSS), and self-administration. Work will be summarized that has shown that certain drugs injected into the mPFC can produce CPP or that lesions of the mPFC can disrupt the development of CPP, that ICSS is obtained with the stimulating electrode placed in the mPFC, and that certain drugs are self administered into the mPFC or that lesions of the mPFC disrupt the peripheral self-administration of certain drugs. However, it has also been shown that the role of the mPFC in reward is not uniform. For example, the mPFC appears to be particularly important for the rewarding actions of cocaine, while it appears not to be important for the rewarding actions of amphetamine. Also, different subareas of the mPFC appear to be differentially involved in the rewarding actions of different drugs. Taken together, the available evidence shows that some drugs can produce reward directly within the mPFC, and that some drugs, even though not having direct rewarding effects within the mPFC, depend on the function of the mPFC for the mediation of their rewarding effects. PMID- 11026493 TI - On excitotoxicity. PMID- 11026492 TI - Atypical antipsychotic-like effect of AMPA receptor antagonists in the rat. AB - Systemic administration of two chemically different AMPA receptor antagonists, GYKI52466, 20mg/kg, and LY326325, 18mg/kg, given subcutaneously, caused a selective suppression of conditioned avoidance response in the rat with preservation of escape behavior. The number of intertrial crosses was not affected and no catalepsy was observed. These experimental results indicate, in principle, an antipsychotic effect of AMPA receptor antagonists with a low liability for extrapyramidal side effects and, consequently, a pharmacological profile consonant with atypical antipsychotic drugs. PMID- 11026494 TI - Effects of kainic acid lesioning on poly(ADP-ribose) polymerase (PARP) activity in the rat striatum in vivo. AB - Poly(ADP-ribose) polymerase (PARP) is activated in glutamate-induced toxicity of neurons in culture (Cosi et al., 1994). Since injection of the excitatory amino acid, kainic acid (KA) into the rat striatum induces a delayed neuronal death, the effects of this in vivo excitotoxin lesioning procedure on striatal PARP activity was investigated. PARP activity was measured in striatal extracts both in the absence ("endogenous" activity) and presence ("total" activity) of exogenously-added fragmented DNA. KA (5nmols/1microl) produced significant and time-dependent changes in striatal PARP activity, compared to saline-injected control animals: no changes at 6h after intrastriatal KA, a 68% and 48% decrease in endogenous and total PARP activity respectively at 12h, a doubling in endogenous PARP activity at 24h, and a 382% and 60% increase in endogenous and total activities at 1 week after KA. PARP cleavage was not detected at any time point. These results suggest a participation of PARP in KA-induced toxicity in the brain in vivo. PMID- 11026495 TI - Drug treatments to reduce excitotoxicity in vivo: a potential for alpha2 adrenoceptor antagonists? AB - It is hypothesized that the locus coeruleus-noradrenergic system controls compensatory and repair mechanisms in the CNS, and that its dysfunction is a critical factor in the progression of central neurodegenerative diseases. Pharmacological activation of locus coeruleus neurons can be achieved with alpha2 adrenoceptor antagonists, and such compounds are protective in vivo in some models of brain injury where excitotoxicity is thought to be a causative factor. To further explore this neuroprotective potential, the effects of a 7-day treatment with the alpha2-antagonists, (+)-efaroxan and (+/-)-idazoxan, were evaluated in rats undergoing a unilateral lesioning of the striatum with the excitotoxin, quinolinic acid. The alpha2-antagonist treatments reduced both the ipsiversive circling response to apomorphine and the deficit of choline acetyltransferase in the lesioned animals. To elucidate the mechanisms underlying this neuroprotective effect, a modulation of the extracellular levels of amino acids within the striatum was investigated using in vivo microdialysis. Intrastriatal injection of quinolinic acid increased taurine and tyrosine levels by 2-2.5 fold, while most other amino acids were not significantly altered; the effect of (+)-efaroxan on these changes is being investigated. Further research is required to identify which of several possible mechanisms is involved in the neuroprotective action of alpha2-antagonists in vivo. PMID- 11026496 TI - Induction of p53 in the glutamate-induced cell death program. AB - Fifteen minute exposure of primary cultures of cerebellar granule cells to micromolar concentrations of glutamate results in apoptotic cell death. Among the intracellular events triggered by glutamate, we identified two transcriptional factors, i.e. the p50 member of the NF-kappaB family and the tumor suppressor phosphoprotein p53, that are apparently linked by a sequential trascriptional program. We found that pretreatment of the cultures with aspirin (ASA), which inhibits NF-kappaB activation, resulted in a complete prevention of glutamate induced p53 immunoreactivity. The same results were obtained pretreating the cells with a specific p53 antisense oligonucleotide. Both ASA and p53 antisense abolished glutamate-induced apoptosis. We also found that two other proteins, the cyclin dependent kinase inhibitor p21 and DNA mismatches repair MSH2, whose encoding genes are well known target of p53, were upregulated by glutamate. On these bases, we propose NF-kappaB, p53, p21 and MSH2 as relevant contributors of the glutamate-induced pro-apoptotic pathway. PMID- 11026497 TI - NMDA receptor dependent and independent components of veratridine toxicity in cultured cerebellar neurons are prevented by nanomolar concentrations of terfenadine. AB - Exposure of cultured neurons to nanomolar concentrations of terfenadine prevented the NMDA receptor-mediated early appearance (30min.) of toxicity signs induced by the voltage sensitive sodium channel activator veratridine. Terfenadine also provided an histamine-insensitive protection against delayed neurotoxicity by veratridine (24h), occurring independently of NMDA receptor activation, while not protecting from excitotoxicity following direct exposure of neurons to glutamate. Terfenadine reduced tetrodotoxin-sensitive inward currents, and reduced intracellular cGMP formation following veratridine exposure. Our data suggest that nanomolar concentrations of TEF may reduce excitatory aminoacid release following neuronal depolarization via a presynaptic mechanism involving voltage sensitive sodium channels, and therefore may be considered as a prototype for therapeutic drugs in the treatment of diseases that involve excitatory aminoacid neurotransmission. PMID- 11026498 TI - The kynurenine pathway of tryptophan degradation as a target for neuroprotective therapies. PMID- 11026499 TI - Possible mediation of quinolinic acid-induced hippocampal damage by reactive oxygen species. AB - Several differences exist between quinolinic acid and N-methyl-D-aspartate (NMDA) in the potency and pharmacology of their neurotoxic actions in the brain, suggesting that quinolinic acid may act by mechanisms additional to the activation of NMDA receptors, possibly involving lipid peroxidation. In the present review, studies are considered which have attempted to determine whether free radicals might contribute to the neuronal damage induced by quinolinic acid. Following Injections into the hippocampus of anaesthetised rats, quinolinic acid induced damage is prevented by melatonin, by an action not blocked by the melatonin receptor blocker luzindole. Deprenyl, but not the non-selective monoamine oxidase inhibitor nialamide, also prevent quinolinic acid-induced damage. In vitro, several groups have shown that quinolinic acid can induce lipid peroxidation of brain tissue The results suggest that free radical formation contributes significantly to quinolinic acid-induced damage in vivo. PMID- 11026500 TI - Electrophysiological effects of exogenous and endogenous kynurenic acid in the rat brain: studies in vivo and in vitro. AB - In this review, recent studies on the electrophysiological effects of de novo synthesized ("endogenous") kynurenic acid (KYNA) are discussed. Endogenous KYNA is normally formed as a byproduct of tryptophan metabolism. Evidence for a physiological role in neuronal excitability has not been strong, in part because brain levels are much lower than the KD of KYNA at the glycine site of the NMDA receptor, where KYNA is thought to exert its most potent effect. The results suggest that, unexpectedly, even low concentrations of endogenous KYNA have physiological consequences. These levels of KYNA reduced the number of hippocampal slices with spontaneous epileptiform discharges after exposure to buffer lacking magnesium. However, effects on evoked responses to single afferent stimuli were not detected. Taken together, the data argue for a potentially important role of endogenous KYNA in suppression of seizure-like activity, and suggest a novel approach to anticonvulsant drug development that could have few side effects. PMID- 11026501 TI - In vivo assessment of kynurenate neuroprotective potency and quinolinate excitotoxicity. AB - Three complementary questions related to the kynurenine pathway and excitotoxicity were addressed in this study: (i) Which extracellular levels of quinolinic acid (QUIN) may be neurotoxic? (ii) Which extracellular levels of kynurenic acid (KYNA) may control excessive NMDA-receptor function? (iii) Can "anti-excitotoxic" levels of KYNA be reached by inhibition of kynurenine-3 hydroxylase (i.e. inhibition of QUIN synthesis and shunts of kynurenine metabolism toward KYNA)? Multifunctional microdialysis probes were used in halothane anaesthetised rats to apply NMDA or QUIN directly to the brain, with or without co-perfusion of KYNA, to record the resulting local depolarisations, and to monitor changes in dialysate KYNA after kynurenine-3-hydroxylase inhibition. QUIN produced concentration-dependent depolarisations with an estimated EC50 (i.e. concentration in the perfusion medium) of 1.22mM. The estimated ED50 for KYNA inhibition of NMDA-responses was 181microM. Kynurenine-3-hydroxylase inhibition (Ro-61-8048, 100mg/kg i.p.) increased dialysate KYNA 11 times (to 33.8nM) but without any reduction of NMDA-responses. These data challenge the notion that extracellular accumulation of endogenous QUIN may contribute to excessive NMDA-receptor activation in some neurological disorders, and the suitability of kynurenine-3-hydroxylase inhibition as an effective anti excitotoxic strategy. PMID- 11026502 TI - Perinatal kynurenine pathway metabolism in the normal and asphyctic rat brain. AB - The kynurenine pathway of tryptophan degradation contains several metabolites which may influence brain physiology and pathophysiology. The brain content of one of these compounds, kynurenic acid (KYNA), decreases precipitously around the time of birth, possibly to avoid deleterious N-methyl-D-aspartate (NMDA) receptor blockade during the perinatal period. The present study was designed to determine the levels of KYNA, the free radical generator 3-hydroxykynurenine (3-HK), and their common precursor L-kynurenine (L-KYN) between gestational day 16 and adulthood in rat brain and liver. The cerebral activities of the biosynthetic enzymes of KYNA and 3-HK, kynurenine aminotransferases (KATs) I and II and kynurenine 3-hydroxylase, respectively, were measured at the same ages. Additional studies were performed to assess whether and to what extent kynurenines in the immature brain derive from the mother, and to examine the short-term effects of birth asphyxia on brain KYNA and 3-HK levels. The results revealed that 1) the brain and liver content of L-KYN, KYNA and 3-HK is far higher pre-term than postnatally; 2) KAT I and kynurenine 3-hydroxylase activities are quite uniform between E-16 and adulthood, whereas KAT II activity rises sharply after postnatal day 14; 3) during the perinatal period, KYNA, but not L-KYN, may originate in part from the maternal circulation; and 4) oxygen deprivation at birth affects the brain content of both KYNA and 3-HK 1 h but not 24h later. PMID- 11026503 TI - Anticonvulsant effects of the 3-hydroxyanthranilic acid dioxygenase inhibitor NCR 631. AB - The kynurenine pathway intermediate 3-hydroxyanthranilic acid (3-HANA) is converted by 3-HANA 3,4-dioxygenase (3-HAO) to the pro-convulsive excitotoxin quinolinic acid. In the present study, the anticonvulsant effect of the 3-HAO inhibitor NCR-631 was investigated in models of chemically- and sound-induced seizures. Administration of NCR-631 i.c.v. at a dose of 300nmol in Sprague-Dawley rats was found to prolong the latency of occurrence of pentylenetetrazole (PTZ) induced seizures. Also systemic pre-treatment with NCR-631 s.c. in N.M.R.I. mice subjected to PTZ-induced seizures provided an increase in the latency until onset of seizures, concomitant with a reduction in the severity of the seizures. However, the anticonvulsant effect of NCR-631 was short lasting (15-30min), and only observed at a dose of 250 mg/kg. A similar dose- and time-dependent anticonvulsant effect of NCR-631 was found in seizure-prone DBA/2J mice following sound-induced convulsions. Hence, the findings show that NCR-631 has anticonvulsant properties against generalized tonic-clonic seizures of different origin, suggesting that it may constitute a useful tool to study the role of kynurenines in various convulsive states. PMID- 11026504 TI - Neurodegeneration and regeneration in the CNS. New roles of heat shock proteins, nitric oxide and carbon monoxide. PMID- 11026506 TI - Neurotrophic factors influence upregulation of constitutive isoform of heme oxygenase and cellular stress response in the spinal cord following trauma. An experimental study using immunohistochemistry in the rat. AB - The influence of brain derived neurotrophic factor (BDNF) or insulin like growth factor-1 (IGF-1) on spinal cord trauma induced carbon monoxide (CO) production and cellular stress response was examined using immunostaining of the constitutive isoform of the hemeoxygenase (HO-2) enzyme and the heat shock protein (HSP 72kD) expression in a rat model. Subjection of rats to a 5 h spinal trauma inflicted by an incision into the right dorsal horn at T10-11 segment markedly upregulated the HO-2 and HSP expression in the adjacent spinal cord segments (T9 and T12). Pretreatment with BDNF or IGF-1 significantly attenuated the trauma induced HSP expression. The upregulation of HO-2 was also considerably reduced. These results show that BDNF and IGF-1 attenuate cellular stress response and production of CO following spinal cord injury which seems to be the key factors in neurotrophins induced neuroprotection. PMID- 11026505 TI - Antioxidant compounds EGB-761 and BN-520 21 attenuate heat shock protein (HSP 72 kD) response, edema and cell changes following hyperthermic brain injury. An experimental study using immunohistochemistry in the rat. AB - Influence of the extract of Gingko biloba (EGB-761) and one of its constituent Gingkolide B (BN-52021) on hyperthermia induced cellular damage and heat shock protein (HSP 72kD) response was examined in a rat model. Rats subjected to 4h heat stress at 38 degrees C in a biological oxygen demand (BOD) incubator (relative humidity 50-55%, wind velocity 20-25cm/sec) resulted in profound edema and cell injury in many parts of the cerebral cortex, hippocampus, cerebellum, thalamus, hypothalamus and brain stem. Immunostaining of HSP 72 kD showed marked upregulation in the damaged and distorted neurons located within the edematous area. Pretreatment with EGB-761 (50mg/kg/day, p.o.) and BN-520 21 (2mg/kg, p.o.) per day for 5 days significantly reduced HSP expression and attenuated cell damage. Our results show that EGB-761 and its component Gingkolide B (BN-52021) has the capacity to reduce edema and cell injury following hyperthermia and this effect of the compound is somehow associated with a reduction in cellular stress response as evidenced with a reduction in HSP expression. PMID- 11026507 TI - Growth hormone attenuates alterations in spinal cord evoked potentials and cell injury following trauma to the rat spinal cord. An experimental study using topical application of rat growth hormone. AB - The influence of exogenous rat growth hormone on spinal cord injury induced alterations in spinal cord evoked potentials (SCEP) and edema formation was examined in a rat model. Repeated topical application of rat growth hormone (20microl of 1microg/ml solution) applied 30min before injury and at 0min (at the time of injury), 10min, 30min, 60min, 120min, 180min, and 240min, resulted in a marked preservation of SCEP amplitude after injury. In addition, the treated traumatised cord showed significantly less edema and cell changes. These observations suggest that growth hormone has the capacity to improve spinal cord conduction and attenuate edema formation and cell injury in the cord indicating a potential therapeutic implication of this peptide in spinal cord injuries. PMID- 11026508 TI - Spinal nerve lesion induces upregulation of constitutive isoform of heme oxygenase in the spinal cord. An immunohistochemical investigation in the rat. AB - The influence of carbon monoxide (CO) on chronic spinal nerve lesion induced spinal cord neurodegeneration was examined using immunohistochemical expression of the constitutive isoform of its synthesising enzyme, hemeoxygenase-2 (HO-2) in a rat model. Spinal nerve lesion at L-5 and L-6 level was produced according to the Chung model of neuropathic pain and rats were allowed to survive for 8 weeks. Sham operated rats, in which the spinal nerves were exposed but not ligated, served as controls. Ligation of spinal nerves in rats resulted in an upregulation of HO-2 expression which was most pronounced in the ipsilateral gray matter of the spinal cord compared to the contralateral side. In these rats, morphological investigations showed distorted neurons, membrane disruption, synaptic damage and myelin vesiculation. Sham operated rats did not show an upregulation of HO-2 expression and the structural changes in the spinal cord were absent. These observations strongly suggest that spinal nerve lesion is associated with an increased production of CO which is somehow contributing to the neurodegenerative changes in the spinal cord, not reported earlier. PMID- 11026509 TI - A new antioxidant compound H-290/51 attenuates nitric oxide synthase and heme oxygenase expression following hyperthermic brain injury. An experimental study using immunohistochemistry in the rat. AB - Influence of a new anti-oxidant compound H-290/51 on expression of nitric oxide synthase (NOS) and heme oxygenase (HO) enzymes responsible for nitric oxide (NO) and carbon monoxide (CO) production, respectively was examined in the CNS following heat stress in relation to cell injury. Exposure of rats to 4h heat stress at 38 degrees C in a biological oxygen demand (BOD) incubator (relative humidity 50-55%, wind velocity 20-25cm/sec) resulted in profound edema and cell injury in many parts of the cerebral cortex, hippocampus, cerebellum, thalamus, hypothalamus and brain stem. Immunostaining of constitutive isoforms of neuronal NOS (nNOS) and HO-2 revealed marked upregulation in damaged and distorted neurons located within the edematous brain regions. Pretreatment with H-290/51 (50 mg/kg, p.o., 30 min before heat stress) significantly reduced the edematous swelling and cell injury and resulted in a marked attenuation of nNOS and HO-2 expression. These observations suggest that upregulation of NOS and HO is associated with cell injury, and the antioxidant compound H-290/51 is neuroprotective in heat stress. PMID- 11026510 TI - Detection of Bence-Jones protein in practice. PMID- 11026511 TI - Lipoprotein(a): what's in a measure? PMID- 11026512 TI - Biochemical investigation of suspected rhabdomyolysis. PMID- 11026513 TI - New diagnostic criteria for diabetes mellitus: are we any further forward? PMID- 11026514 TI - Genetic and biochemical markers for Alzheimer's disease: recent developments. PMID- 11026515 TI - Measurement of endothelin: clinical and research use. PMID- 11026516 TI - Total homocysteine measurement in clinical practice. PMID- 11026517 TI - Effect of vitamin E supplementation on circulating cell adhesion molecules pre- and post-coronary angioplasty. AB - The soluble adhesion molecules P-selectin (sP-selectin) and intercellular adhesion molecule-1 (sICAM-1) are derived from platelets and endothelial cells. Circulating concentrations of these soluble adhesion molecules are raised in patients with atherosclerosis and following percutaneous transluminal coronary angioplasty (PTCA). We have investigated the effects of vitamin E supplements (800 IU/day) on circulating plasma ICAM-1 and P-selectin levels pre- and post PTCA. Patients, randomized to group, were pre-treated with vitamin E or placebo (soybean oil) for 1 month before routine PTCA. Plasma sICAM-1 and sP-selectin were measured by enzyme-linked immunosorbent assay on blood taken immediately pre and post-PTCA. Total protein and alpha-tocopherol were measured on the same samples. Plasma alpha-tocopherol concentrations increased in patients receiving vitamin E: 19.1 (1.5) [mean (standard error of the mean, SEM)] mg/mL post-PTCA versus 13.9 (0.6) mg/mL pre-PTCA (n=23; P<0.01). Plasma sP-selectin and sICAM-1 levels were not significantly increased following PTCA in the vitamin E group. Pre-angioplasty mean (SEM) plasma sP-selectin concentration in the vitamin E group was 8.83 (0.97) ng/mg protein; the corresponding mean post-angioplasty value was 9.34 (0.89) ng/mg protein (P=0.85). The mean (SEM) pre-angioplasty sICAM-1 concentration in this group was 2.18 (0.24) ng/mg protein, and was 2.20 (0.23) ng/mg protein following angioplasty (P = 0.84). In the placebo group (n = 24) there was a significant increase in mean (SEM) sP-selectin concentration following angioplasty, from 7.48 (0.73) to 9.70 (0.78) ng/mg protein (P<0.05). The change (mean, SEM) in plasma sP-selectin concentration following angioplasty was significantly greater for the placebo group [2.22 (0.50) ng/mg protein] than for the group receiving vitamin E [0.50 (0.50) ng/mg protein] (P<0.02). This difference remained significant (P<0.05) even after adjustment for pre angioplasty P-selectin concentrations. Mean (SEM) plasma sICAM-1 concentrations remained unchanged following angioplasty [pre-angioplasty: 2.16 (0.20) ng/mg protein; post-angioplasty: 1.97 (0.13) ng/mg protein]. Vitamin E may therefore limit platelet or endothelial activation during PTCA. PMID- 11026518 TI - Hyperfibrinogenaemia did not improve after treating hyperglycaemia in Chinese type 2 diabetic patients. AB - Diabetic patients have higher cardiovascular risk than non-diabetic subjects, and fibrinogen has been reported to be one of the independent predictors for diabetic vascular complications. We examined 101 subjects (24 men, 77 women) who were newly diagnosed with type 2 diabetes. Thirty non-diabetic subjects were recruited as controls. The mean (standard deviation, SD) baseline fibrinogen concentrations in men and women were 3.87 (1.17) and 3.42 (1.00) g/L, respectively (not significantly different). Of these 101 subjects, 70 were treated with diet alone and 31 were treated with oral agents. After a mean (SD) follow-up period of 2.38 (0.63) years, there was an 18.5% increase in the fibrinogen concentration from 3.53 (1.06) g/L at baseline to 3.97 (1.07) g/L at follow-up (P < 0.001). The normal subjects had a mean (SD) follow-up period of 1.17 (0.38) years, and showed no significant change in their fibrinogen concentrations from 3.49 (0.90) g/L at baseline to 3.15 (0.61) g/L at follow-up. In the 101 diabetic subjects, only changes in plasma triglyceride correlated with the changes in fibrinogen concentration. Using multivariate analysis with age, sex, duration of diabetes, baseline and changes in body mass index, blood pressure, glycaemic and lipid parameters as independent variables (R2 = 0.106, F= 6.840, P = 0.011), triglyceride (beta = 0.282, P = 0.011) was identified as the only independent variable that predicted the changes in the fibrinogen concentrations. Improved glycaemic control was not accompanied by a reduction in plasma fibrinogen concentration. PMID- 11026519 TI - Assessment of iron status in cystic fibrosis patients. AB - We investigated the use of soluble transferrin receptor (sTfR), together with more traditional measurements such as iron, transferrin and ferritin, to assess iron status in 70 adult cystic fibrosis patients. sTfR concentration was measured using particle-enhanced immunonephelometry. We found that 69% of subjects as determined by transferrin saturation had iron deficiency, but only 29% as determined by sTfR and 11% as determined by ferritin. We found a significant correlation between C-reactive protein (CRP) and both ferritin (r = 0.38, P = 0.0001) and transferrin saturation (r = -0.54, P < 0.0001). In addition, because the CRP concentration was elevated in 64% of subjects, we feel that the transferrin saturation was overestimating, and the ferritin underestimating, iron deficiency in these patients. The sTfR concentration, on the other hand, is unaffected by the acute-phase response and was therefore thought to be the most useful test for detecting iron deficiency in this group of patients. PMID- 11026520 TI - Near-patient test for nicotine and its metabolites in saliva to assess smoking habit. AB - Smoking is a major risk factor for periodontal disease, causing bone and tooth loss. Effective management of many oral conditions requires accurate information about smoking. We describe a disposable, 10-min, near-patient saliva test to monitor cigarette smoking. A plastic device contains the dried reagents to measure nicotine and its metabolites by a colorimetric assay. The device can be used to give a semi-quantitative assessment of tobacco consumption by observing a colour change and comparing it to a reference chart. Alternatively, the test can be quantified by measuring the light absorbance with a colorimeter and determining the concentration of nicotine metabolites by reference to a cotinine standard. This method correlates with reported daily cigarette consumption (r=0.62, P<0.0001) and nicotine yield of cigarettes smoked (r=0.35, P<0.01) and compares favourably with cotinine, as measured independently by radioimmunoassay (r=0.57, P<0.005). The method provides a simple-to-use, inexpensive way of monitoring tobacco consumption in dental surgeries. PMID- 11026521 TI - Direct polymerase chain reaction from whole blood without DNA isolation. AB - Blood and other animal fluids contain a variety of substances that inhibit the polymerase chain reaction (PCR), so that isolation of DNA is generally necessary prior to PCR. We have developed a novel reagent cocktail that effectively suppresses these inhibitory substances and makes DNA isolation from blood unnecessary for PCR. When this reagent was included in the PCR mixture, DNA fragments of the beta-globin gene could be efficiently amplified directly from human blood samples treated with various anticoagulants or PCR-inhibitory substances. We confirmed the usefulness of this cocktail by examining a large number of blood samples with various PCR primer sets. In addition to fresh blood, this method enabled PCR amplification from blood samples stored at 4 degrees C, 20 degrees C or -80 degrees C for a minimum of 1 year. PMID- 11026522 TI - Alpha-fetoprotein variants in a case of pancreatoblastoma. AB - We describe a 6-year-old boy with histologically confirmed pancreatoblastoma and a markedly elevated serum alpha-fetoprotein (AFP) concentration. Due to local tumour invasion, cytotoxic chemotherapy was given to debulk the tumour before attempting surgical resection. Serial serum AFP concentrations were measured on this patient. During chemotherapy there was a > 95% fall in total AFP. Tumour specific variants of AFP, detected by isoelectric focusing, also disappeared during chemotherapy but recurred when chemotherapy was withdrawn. It is suggested that although there was no change in overall size of the tumour, as assessed by various imaging techniques, the changes in serological markers may indicate that the treatment did in fact cause considerable tumour necrosis, AFP and its variants may be useful markers of tumour response in patients with pancreatoblastoma. The expression of AFP and its variants in pancreatoblastoma may be related to the embryonic origin of the pancreas. PMID- 11026523 TI - Moderately low vitamin B12 does not compromise transmethylation in adults on a free diet: implications for assessment of vitamin B12 status. AB - Lower reference limits for vitamin B12 are often defined solely in relation to haematological criteria. This may be misleading and there is evidence indicating that biochemical anomalies should also be considered. In 50 patients we measured creatine, as the major product of B12-mediated remethylation, to see if this helps the definition of the B12 reference interval and to investigate the possible effect of low B12 on essential transmethylation. Vitamin B12 values were grouped into six fractiles covering the range 50-500 ng/L; the corresponding creatine results were assessed by analysis of variance giving F = 0.94 and a significance of 0.466. Although no correlation between B12 and creatine was found, and therefore no obvious effect on transmethylation of low B12 concentration, this must be interpreted with respect to methionine availability. Other factors indicate that biochemical perturbations should be taken into account when defining vitamin B12 reference intervals. PMID- 11026524 TI - Analysis of urine for drugs of abuse using mixed-mode solid-phase extraction and gas chromatography-mass spectrometry. AB - A method for the simultaneous analysis of urine for the major drugs of abuse is described. The analytical procedure uses solid-phase extraction (SPE), gas chromatography-mass spectrometry (GC-MS) and a semi-automated identification process. It allows simultaneous extraction, derivatization and analysis of acidic, neutral and basic drugs from urine. Urine samples were subjected to enzymatic hydrolysis followed by SPE using Bakerbond narc-2 columns. The eluant was selectively derivatized with N-methyl-bis-trifluoroacetamide (MBTFA) and N methyl-N-trimethylsilyltrifluoroacetamide + 1% trimethylchlorosilane. Analysis was performed using a GC-MS system operating in full scan mode. A simple macro programme was written to enhance the mass spectra identification capabilities of the MS software by producing extracted ion chromatograms (EIC) for the drugs of interest. Once a suspect compound was indicated by EIC, the mass spectrum of the compound was searched manually against reference libraries for positive identification and the retention time checked against that of the standard. This procedure has increased both the amount and the reliability of information given to clinicians without increasing the cost per sample. The system has been in routine operation for 24 months, processing up to 40 urine samples per day, with a usual turn-around time of 48 h. PMID- 11026525 TI - Reference values of plasma lipoprotein (a) in newly classified apolipoprotein(a) phenotype groups in the Japanese population. AB - We tried to establish the reference values of plasma lipoprotein (a) [Lp(a)] concentration in phenotype groups of apoliprotein(a) [apo A] classified by a new criterion. Lp(a) concentration was determined by latex agglutination immunoassay, and apo A was analysed by electrophoresis in sodium dodecyl sulphate polyacrylamide gel and a Western blotting technique. According to the relative mobility to the apo B-100 band, apo A was classified into 11 isoforms, i.e. F, B, and S1-S9, and the phenotype was defined by their apparent combination. The frequency ratio of single-band versus double-band was approximately 2:1. In 382 cases of single-band, the most frequent phenotype was S5 (24.3%), followed by S4 (17.3%), S6 (15.4%) and S3 (14.4%). In 181 cases of double-band, S5/S6 phenotype was observed most frequently (12.2%). followed by S4/S5 (10.5%) and S3/S6 (7.2%). The reference value was determined between antilogs of the mean +/- 1.96 standard deviation by logarithmic transformation of all observed values for individual phenotype cases. These results suggest that the reference values shown to be variable with apo A phenotypes should be useful for evaluating Lp(a) values in diagnosis of atherosclerosis. PMID- 11026526 TI - Capillary isotachophoretic analysis of serum lipoproteins using a carrier ampholyte as spacer ion. AB - We have developed a novel analytical method for serum lipoproteins using a commercially available capillary electrophoresis apparatus, BioFocus 3000 (Bio Rad Laboratories Co Ltd, USA). The analytical principle is isotachophoresis (ITP), using a carrier ampholyte, BioLyte 7/9, as a spacer ion. The method allows a much higher resolution of lipoproteins than of amino acid mixtures. Serum lipoproteins are normally separated into 13-15 peaks, including some shoulder peaks. The reproducibility of repeated analysis within a day was relatively good with the coefficient of variation within the range 0.9-1.1%. VLDL, LDL and HDL prepared by discontinuous density ultracentrifugation could be further separated by capillary ITP. This high-resolving ability of our method enabled detection of small amounts of abnormal lipoprotein species. For example, small dense LDL, which is thought to be an atherogenic lipoprotein, could be detected within the LDL group peak. Moreover, an abnormal HDL, apolipoprotein E-rich HDL, was also detected by a single analysis. These findings suggest that our capillary ITP method is a useful means for detailed analysis of lipoproteins and thus for clinical diagnosis of hyperlipoproteinaemic subjects. PMID- 11026527 TI - Differences in circulating concentrations of total, free and bound leptin relate to gender and body composition in adult humans. AB - We describe a radioimmunoassay (RIA) for total leptin and a gel filtration procedure for the separation of free and bound leptin in human serum. The RIA, based on a locally prepared antibody, has a minimum detection limit of 0.9 ng/mL, a working range (CV < 10%) of 2.5-50 ng/mL, inter-assay precision of 10.2, 7.2 and 8.9%CV at 7.9, 15.4 and 30.0 ng/mL, respectively, 94% recovery of exogenous leptin (range 81.1-120.6%), exhibited parallelism and demonstrated no significant cross-reactivity or interferences. A difference plot of results from this method and those from a commercially available kit (Linco Research) demonstrated satisfactory agreement up to concentrations of 50 ng/mL total leptin, with no significant bias. A gender-dependent correlation was obtained between body mass index (BMI) and total leptin (r = 0.91, P<0.001, n = 75 for men; r = 0.79, P<0.001, n = 72 for women), with women having higher leptin concentrations than men for any given BMI. Gel filtration studies (inter-assay precision: 4.7%CV, n = 18) demonstrated that a variable fraction (between 10% and 40%) of total leptin in serum was bound with high affinity (Keq = 1.0-1.45 x 10(9) L/mol) to a non albumin, non-lipid macromolecule. Binding affinities were found to be similar irrespective of gender or fat mass. A significant positive correlation between free or bound leptin concentrations and BMI was obtained for both men and women (r = 0.87-0.94); free and bound leptin concentrations were also significantly higher in women (P<0.01) than in men for any given BMI, and higher in obese (P<0.01) than in lean individuals. We conclude that leptin 'resistance' associated with obesity cannot be accounted for by reduced free leptin concentrations in serum and that the methods described are suitable for the investigation of total, free and bound leptin for both clinical and research purposes. PMID- 11026528 TI - Measurement of plasma 5-hydroxyindoleacetic acid in carcinoid disease: an alternative to 24-h urine collections? PMID- 11026529 TI - Severe hyperammonaemia in a previously healthy teenager. PMID- 11026530 TI - Cider potomania. PMID- 11026531 TI - 'Glucose meter hypoglycaemia': often a non-disease. PMID- 11026532 TI - Cardiac marker testing. PMID- 11026533 TI - Thyroid function tests in acutely ill patients. PMID- 11026534 TI - Thyroid function tests in acutely ill patients. PMID- 11026535 TI - The structure of an HIV-1 specific cell entry inhibitor in complex with the HIV-1 gp41 trimeric core. AB - The three-dimensional structure of the complex between an HIV-1 cell-entry inhibitor selected from screening a combinatorial library of non-natural building blocks and the central, trimeric, coiled-coil core of HIV-1 gp41 has been determined by X-ray crystallography. The biased combinatorial library was designed to identify ligands binding in nonpolar pockets on the surface of the coiled-coil core of gp41. The crystal structure shows that the non-peptide moiety of the inhibitor binds to the targeted cavity in two different binding modes. This result suggests a strategy for increasing inhibitor potency by use of a second-generation combinatorial library designed to give simultaneous occupancy of both binding sites. PMID- 11026536 TI - Template-constrained cyclic peptide analogues of somatostatin: subtype-selective binding to somatostatin receptors and antiangiogenic activity. AB - Beta-turns are a common secondary structure motif found in proteins that play a role in protein folding and stability and participate in molecular recognition interactions. Somatostatin, a peptide hormone possessing a variety of therapeutically-interesting biological activities, contains a beta-turn in its bioactive conformation. The beta-turn and biological activities of somatostatin have been succesfully mimicked in cyclic hexapeptide analogues. Two novel, structured, non-peptidic molecules were developed that are capable of holding the bioactive tetrapeptide sequence of somatostatin analogues in a beta-turn conformation, as measured by somatostatin receptor (SSTR) binding. Template constrained cyclic peptides in which the ends of the -Tyr-D-Trp-Lys-Val tetrapeptide were linked by scaffolds based on either an N,N'-dimethyl-N,N' diphenylurea or a substituted biphenyl system (DJS631 and DJS811, respectively), bound selectively to mouse SSTR2B and rat and human SSTR5 with affinities as high as 1 nM. DJS811, at a dose of 3 mg/kg/day, was shown in a mouse Matrigel model to inhibit angiogenesis to a level of 79%. The development of structured turn scaffolds allows beta-turn sequences to be contained in the context of a compact structure, with less peptidic nature and potentially greater bioavailability than cyclic hexapeptides. These systems can be used to study the determinants of beta turn formation, as well as to probe the importance of turn sequences occurring in molecular recognition interactions. The antiangiogenic activity of DJS811 suggests that it may have antitumor activity as well. In addition, because SSTR2 is overexpressed on many types of tumors, DJS631 and DJS811 may be useful in the development of agents for tumor imaging or the radiotherapy of cancer. PMID- 11026537 TI - Design and synthesis of novel potent antinociceptive agents: methyl-imidazolyl N acylhydrazone derivatives. AB - This paper describes recent results of design, synthesis and pharmacological evaluation of new N-heterocyclic functionalized N-acylhydrazone compounds, belonging to the 2-methyl-imidazolyl-3-acylhydrazone class (4a-e). These compounds were planned by applying the molecular simplification strategy to propose the structural modifications on the previously described functionalized imidazo [1,2-a]pyridine 3-acylhydrazone series (2), which presented an important analgesic profile. This new series (4) was synthesized in order to investigate the possible pharmacophoric contribution of the N-heteroaromatic ring and N acylhydrazone moieties to the analgesic activity. Compounds 4a-b are the most potent antinociceptive agents from this series. PMID- 11026538 TI - Synthesis and enzymatic evaluation of five-membered iminocyclitols and a pseudodisaccharide. AB - Described here are the synthesis of five-membered iminocyclitols with galacto configuration and a pseudodisaccharide, and their inhibitory activities against beta-galactosyltransferase, beta-galactosidase and alpha-mannosidase. PMID- 11026540 TI - Glucose-6-phosphatase catalytic enzyme inhibitors: synthesis and in vitro evaluation of novel 4,5,6,7-tetrahydrothieno[3,2-c]- and -[2,3-c]pyridines. AB - The discovery of the first class of potent glucose-6-phosphatase catalytic site inhibitors, substituted 4,5,6,7-tetrahydrothieno[3,2-c]- and -[2,3-c]pyridines, is described. Optimisation of this series involved solution phase combinatorial synthesis and very potent compounds were prepared with IC50 values down to 140 nM. The structure activity relationship (SAR) of these compounds indicates that: a tetrahydrothieno[3,2-c]pyridine core ring system and the isomeric [2,3-c] system are equipotent and much better than the corresponding benzo analogue, 1,2,3,4-tetrahydro-isoquinoline. The 4-substituent of the tetrahydrothieno[3,2 c]pyridine ring has to be a phenyl group, optionally substituted with a lipophilic 4-substituent, such as trifluoromethoxy or chloro. The 5-substituent of the tetrahydrothieno[3,2-c]pyridine ring has to be a substituted benzoyl; anisoyl and (E)-3-furan-3-ylacryloyl are the best of the investigated groups. Substitution in the benzoyl ortho position seems to be forbidden, whereas substitution in the meta position is tolerated only if a methoxy para substituent is present. These SAR findings were parallel to those obtained in the 4,5,6,7 tetrahydrothieno[2,3-c]pyridine system. Enantioselectivity in enzyme recognition was observed and the activity resided in all cases only in one of the enantiomers. PMID- 11026539 TI - Design, synthesis and biological evaluation of pyridine-phenylpiperazines: a novel series of potent and selective alpha1a-adrenergic receptor antagonist. AB - Beginning from the screening hit and literature alpha1-adrenergic compounds, a hybridized basic skeleton A was proposed as the pharmacophore for potent and selective alpha1a-AR antagonists. Introduction of a hydroxy group to increase the flexibility afforded B which served as the screening model and resulted in the identification of the second-generation lead 1. Using the Topliss approach, a number of potent and selective alpha1a-AR antagonists were discovered. In all cases, binding affinity and selectivity at the alpha1a-AR of S-hydroxy enantiomers were higher than the R-hydroxy enantiomers. As compared to the des hydroxy analogues, the S-hydroxy enantiomers displayed comparable potency and better selectivity at alpha1a-AR. The S-hydroxy enantiomer 17 (Ki = 0.79 nM; alpha1b/alpha1a = 800; alpha1d/alpha1a = 104) was slightly less potent but much more selective at alpha1a-AR than tamsulosin (Ki = 0.13 nM, alpha1b/alpha1a = 15, alpha1d/alpha1a = 1.4). Compound 17 displayed higher selectivity in inhibiting rat prostate contraction over rat aorta contraction and also exhibited a higher degree of uroselectivity than tamsulosin in the anesthetized dog model. PMID- 11026541 TI - Selective boron-containing thrombin inhibitors--X-ray analysis reveals surprising binding mode. AB - Based on the structural comparison of the S1 pocket in different trypsin-like serine proteases, a series of Boc-D-trimethylsilylalanine-proline-boro-X pinanediol derivatives, with boro-X being different amino boronic acids, have been synthesized as inhibitors of thrombin. Among the novel compounds, a number of derivatives were synthesized which appeared to have side-chain variants too big to fit into the S1 pocket. Nevertheless, these compounds inhibited thrombin in the nM range. The X-ray structure of one of these inhibitors bound to the active side of thrombin reveals that a new binding mode is responsible for these surprising results. PMID- 11026543 TI - The conformation and activity relationship of benzofuran type of angiotensin II receptor antagonists. AB - As a continuing effort to establish the structure and activity relationship in a benzofuran type of angiotensin II antagonist, we synthesized various regioisomers and performed a series of QSAR analyses. The conformational analyses of target isomers were carried out using molecular mechanics and fine-tuned using the information from the NMR NOE experiment. The conformations of compounds with a good binding activity are quite similar to that of DuP753, a prototype of AII antagonist, suggesting that these compounds also bind to the same site of AII receptor. We then studied the compounds with a varied length of the hydroxyl group bearing side chain to find out the optimum distance between the hydroxyl group and the imidazole ring. The CoMFA with these compounds gave acceptable statistical measures (cross-validated r2 and conventional r2 to be 0.881 and 0.974, respectively) and the map was well consistent with the previously proposed pharmacophore. PMID- 11026542 TI - Computer-assisted design, synthesis and biological evaluation of novel pyrrolyl heteroaryl sulfones targeted at HIV-1 reverse transcriptase as non-nucleoside inhibitors. AB - Three pyrrolyl heteroaryl sulfones (ethyl 1-[(1H-benzimidazol-2(3H)one-5 yl)sulfonyl]-1H-pyrrole-2-carboxyla te, ethyl 1-[(1H-benzimidazol-5(6) yl)sulfonyl]-1H-pyrrole-2-carboxylate and ethyl 1-[(1H-benzotriazol-5(6) yl)sulfonyl]-1H-pyrrole-2-carboxylate) were designed as novel HIV-1 reverse transcriptase non-nucleoside inhibitors using structure-based computational methods. Although these compounds were inactive in the cell-based assay, they inhibited the target enzyme with micromolar potency (IC50s = 2 microM, 3 microM and 9 microM, respectively). PMID- 11026544 TI - Studies on anti-Helicobacter pylori agents. Part 2: new cephem derivatives. AB - The synthesis and optimization of the anti-Helicobacter pylori activity of a novel series of cephem derivatives are described. Introduction of thio heterocyclic groups containing N- and S-atoms to the 3-position and phenyl or thienyl acetamido groups to the 7-position of the cephem nucleus dramatically improved the activity. From this series of derivatives, compound 13i was found to have extremely potent in vitro anti-H. pylori activity, superior therapeutic efficacy compared to AMPC and CAM, no cross-resistance between CAM or MNZ and low potential for causing diarrhea due to instability to beta-lactamase. PMID- 11026545 TI - Noncompetitive antagonist-binding sites of rat and housefly gamma-aminobutyric acid receptors display different enantiospecificities for tert butyl(isopropyl)bicyclophosphorothionate. AB - The enantiomers of 4-tert-butyl-3-isopropyl-2,6,7-trioxa-1-phosphabicyclo[2.2.2 ]octane 1-sulfide (TBIPPS) were prepared in nine steps from diethyl tert butylmalonate, and their abilities to compete with [3H]1-(4-ethynylphenyl)-4-n propyl-2,6,7-trioxabicyclo[2.2.2 ]octane (EBOB), a noncompetitive antagonist of ionotropic gamma-aminobutyric acid (GABA) receptors, at their binding site were investigated using rat brain and housefly head membranes. The (S)-(-)-isomer of TBIPPS (IC50 = 398 nM) was more potent than was the (R)-(+)-isomer of TBIPPS (IC50 = 1220 nM) in rat receptors, while the potencies of (S)-TBIPPS 104 nM) and (R)-TBIPPS (IC50 = 94.4 nM) in housefly receptors were almost the same. The different enantiospecificities of rat and housefly receptors indicate that the three-dimensional structure of the binding site might be different between these receptors. In a region of the rat binding site there might be a steric bulk that interacts less favorably with (R)-TBIPPS than with (S)-TBIPPS, while in the corresponding region of the housefly binding site there might not be such a steric bulk that leads to specificity for these compounds. PMID- 11026546 TI - Pyrazolo-triazoles as light activable DNA cleaving agents. AB - In view of the continuous interest in new DNA cleaving compounds, both for the development of new therapeutic agents and for the possible use as reagents in nucleic acids research, a few pyrazolo[3,4-d][1,2,3]triazole derivatives have been obtained and investigated for their antiproliferative activity and capability to cleave DNA, after light-activation. A possible in situ activation, i.e. in neoplastic tissues, of less cytotoxic derivatives, may lead to potential antitumor compounds endowed with high therapeutic indexes. PMID- 11026547 TI - Syntheses and biological evaluation of new glyco-modified angucyclin-antibiotics. AB - The synthesis of novel aquayamycin-derived angucycline antibiotics 13a-d has been achieved. Glycosylation of aquayamycin (6) using 2-selenoglycosyl acetate 7 as glycosyl donor proceeded in excellent yield but attempts to reductively remove the selenyl group led to rearrangement or further aromatization of the aglycon. As a consequence of these results, it became possible to prepare urdamycinone B (10) starting from aquayamycin (6). In addition, silyl protected D-olivals 12a,b were attached to the C-glycoside domain of aquayamycin (6) under protic conditions. As expected, the hydroxy and phenol groups of the benz[a]anthracene framework of 6 did not react under the glycosylation conditions employed. Stepwise removal of the silyl protecting group starting with tetrabutyl ammonium fluoride followed by use of the HF/pyridine complex suppressed a possible rearrangement of the aglycon and successfully terminated the sequence. The new angucycline-antibiotics 13a and 13b are some of the most potent xanthine oxidase inhibitors known and show cytotoxic activity with ED50-values in the range of 12.6-2.9x 10(-6) M. PMID- 11026548 TI - 2-Amino-Benzo[d]isothiazol-3-one derivatives: synthesis and assessment of their antiplatelet/spasmolytic effects. AB - We describe a series of 2-amino-benzo[d]isothiazol-3-one derivatives (2-8), which were synthesized and screened in vitro for inhibition of platelet aggregation and for their spasmolytic activity, with the awareness that the development of antiplatelet agents with additional vasodilation activity could be beneficial in the treatment of various vaso-occlusive disorders. The tested compounds show a powerful antiplatelet activity and various modifications resulted in molecules possessing antiaggregating effects as well as spasmolytic actions. PMID- 11026549 TI - Potential antitumor agents. Part 29(1): synthesis and potential coanthracyclinic activity of imidazo[2,1-b]thiazole guanylhydrazones. AB - This paper reports the synthesis of new imidazo[2,1-b]thiazole guanylhydrazones which were tested as potential antitumor agents. Three of these derivatives (those bearing a 3- or 4-nitrophenyl group) were the most potent and one of these showed a mild effect as CDK1 inhibitor. These same three derivatives were also tested as positive inotropic agents and two of them were more potent than amrinone at 10(-5) M. These two guanylhydrazones could be useful coanthracyclinic agents. PMID- 11026550 TI - C-nucleoside analogues of furanfurin as ligands to A1 adenosine receptors. AB - Furanfurin (2-beta-D-ribofuranosylfuran-4-carboxamide) derivatives and analogues were synthesized and their affinity for adenosine receptors was determined. The agonistic behavior of furanfurin against A1 receptors is preserved only when the furan ring is substituted with isosteric pentatomic ring systems such as oxazole, thiazole or thiophene, and the carboxamide group is unsubstituted. Replacement of the hydrogen atoms of the carboxamide group with alkyl, cycloalkyl or arylalkyl groups generates compounds endowed with moderate antagonistic activity. PMID- 11026551 TI - Comparison of three gamma-turn mimetic scaffolds incorporated into angiotensin II. AB - Rigidification of peptides by cyclization and iterative incorporation of well defined secondary structure mimetics constitutes one approach to the design of non-peptidergic structures with better defined conformations. We herein present the synthesis of a potential gamma-turn mimetic scaffold, and its incorporation in the 3-5 position of angiotensin II. Two analogues of angiotensin II (Ang II) incorporating this 1,3,5-trisubstituted benzene gamma-turn scaffold were synthesized. Evaluation of the compounds in a radioligand binding assay showed that they lacked affinity to the AT1 receptor. To rationalize these results a geometrical and electrostatical comparison with Ang II analogues encompassing a bicyclic scaffold that delivered inactive pseudo peptides and an azepine scaffold producing highly active ligands was made. This analysis did not provide a clear rationale for the inactivity of the benzene gamma-turn scaffolds. PMID- 11026552 TI - Control of gene expression by growth hormone in liver: key role of a network of transcription factors. AB - Growth hormone (GH) controls gene expression in liver. Recent work suggests that this can result in part from the stimulation by GH of the synthesis of liver specific transcription factors, one of which is HNF-6. The liver-specific factors HNF-4 and C/EBP alpha respectively stimulate and inhibit transcription of the hnf 6 gene. Upon GH stimulation, the affinity of HNF-4 for the hnf 6 promoter is increased and the binding of C/EBP alpha is decreased. GH therefore controls hnf 6 by a combination of stimulatory and derepressive mechanisms. On the other hand, HNF-6 stimulates transcription of the hnf 3beta and hnf 4 genes, the stimulation of hnf 4 resulting most likely from the GH-induced increase in HNF-6 concentration. We conclude that in liver GH is likely to control the synthesis of a whole set of proteins whose genes are regulated by a GH-sensitive network of transcription factors, which regulate each other by feed-back and autoregulatory loops. PMID- 11026553 TI - Demonstration of 2,3,7,8-tetrachlorodibenzo-p-dioxin attenuation of P450 steroidogenic enzyme mRNAs in rat granulosa cell in vitro by competitive reverse transcriptase-polymerase chain reaction assay. AB - We investigated the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), in prepubertal (PP) and adult (A) rat granulosa cells (GC) in vitro by examining the changes in estrogen secretion, aromatase enzyme activity and mRNAs for steroidogenic enzymes P450scc, 3beta-HSDI, P450arom; and for components of the AHR signaling pathway-CYP1A1, aromatic hydrocarbon receptor (AHR), and the AHR nuclear translocator protein (ARNT). In PP and A rat GC, TCDD (3.1 nM) reduced estrogen secretion at 48 h without altering aromatase enzyme activity. Addition of FSH (50 ng/ml) increased aromatase activity in GC with or without TCDD. FSH induced aromatase activity was significantly reduced by TCDD (3.1 nM) at 48 h. Semi-quantitative RT-PCR showed a significant increase in CYP1A1 mRNA both at 24 and 48 h with TCAP, while a significant reduction in P450scc and P450arom mRNA was observed with competitive RT-PCR. All steroidogenic enzyme mRNAs were significantly lower in adults than in PP GC. We conclude that in rat GC, TCDD modulates the level of cytochrome P450 enzymes involved in the steroid biosynthetic cascade. This effect may be attributable to AHR interaction with dioxin-responsive elements present in the genes encoding these enzymes. PMID- 11026554 TI - Regulation of estrogen receptor levels by ligand-induced release of compound(s) in MCF-7 cells. AB - In MCF-7 cells, estradiol (E2) and pure antiestrogens (AEs) decrease estrogen receptor alpha (ER) levels, while AEs with partial estrogenic activity lead to ER alpha accumulation. Using immunocytochemistry, we found that cells pre-exposed to one of such ligands, when plated with untreated cells, led to similar ER changes in the latter. Conditioned media (CMs) prepared from stimulated cells displayed identical regulatory effects even after strong dilution; they also modulated ERE dependent transcriptional activity. Evaluation of residual ligand concentrations in CMs rejected the possibility of a major interference of the former. Cycloheximide, which inhibits E2-induced down-regulation, failed to block the influence of CM(E2) in agreement with this view. DCC-treatment of CMs abrogated their effects, suggesting the release of hydrophobic compound(s) which regulate ER and/or amplify the effect of extremely low amounts of residual ligands. Such a release appears independent of ER since CMs from MDA-MB-231 cells (ER-negative) were effective as their autologous media on MCF-7 cells. PMID- 11026555 TI - Misfolded vasopressin V2 receptors caused by extracellular point mutations entail congential nephrogenic diabetes insipidus. AB - Vasopressin V2 receptor mutants from three different patients with congenital nephrogenic diabetes insipidus phenotypes were investigated after expression in COS cells. The amino acid exchanges within the human V2 receptor are located in the second extracellular loop (T204N, Y205C and V206D). Confocal microscopy showed that all receptor mutants were strongly expressed but mainly located within the cell. Residual binding capacity for the antidiuretic hormone arginine vasopressin (AVP) could only be detected for the T204N mutant and was 10-fold lower than for the wild-type receptor. Stimulation of transfected cells with 1 microM AVP showed that the T204N mutant was able to activate the adenylyl cyclase pathway. In contrast, the Y205C mutant was almost inactive and stimulation of the V206D mutant increased the cAMP accumulation only slightly. Dose dependent stimulation of cells expressing the T204N mutant with AVP and with the therapeutic AVP analogue 1-deamino[D-Arg8]vasopressin (dDAVP) revealed that AVP was 50-fold more potent than dDAVP. This indicates that the ligand binding selectivity of the T204N mutant has changed as compared with the wild-type receptor where AVP is only 2.3-fold more potent than dDAVP. Despite its defects in membrane localization, ligand binding affinity and selectivity, the T204N receptor could be activated with high concentrations of dDAVP. Our results indicate that in cases of congenital nephrogenic diabetes insipidus with residual V2 receptor activities the use of antidiuretic drugs, such as dDAVP, might be beneficial for patients. PMID- 11026556 TI - Alterations in retinoid X receptor-alpha expression contribute to cell-type dependent differences in thyroid hormone regulation of malic enzyme transcription. AB - Triiodothyronine (T3) stimulates a marked increase (> 40-fold) in transcription of the malic enzyme gene in chick embryo hepatocytes (CEH), but has no effect on malic enzyme transcription in chick embryo fibroblasts (CEF) that express nuclear T3 receptors (TR) at levels which are similar to those of CEH. Heterodimerization of the TR with other nuclear proteins is a potential mechanism for the regulation of T3 action. For example, heterodimers of retinoid X receptors (RXR) and TR bind to T3 response elements (T3RE) with higher affinity and modulate transcription more effectively than TR homodimers. In the present report, we investigated the role of RXR in mediating differences in T3 responsiveness of the malic enzyme gene between CEH and CEF. Data from gel mobility shift analyses demonstrated that endogenous TRs from CEH and CEF bind to the major T3RE of the malic enzyme gene primarily as heterodimers with RXR alpha or a protein highly related to RXR alpha. The total binding activity of RXR alpha/TR complexes in CEF was decreased relative to that observed in CEH. Cell-type dependent differences in RXR alpha/TR complex formation were greater in cells incubated in the presence of T3 because T3 treatment increased RXR alpha/TR binding activity in CEH but had no effect on RXR alpha/TR binding activity in CEF. Decreased RXR alpha/TR complex formation in CEF relative to CEH was associated with a reduction in the abundance of RXR alpha protein and RXR alpha mRNA in the former cell-type. Expression of exogenous RXR alpha in CEF increased the T3 responsiveness of the malic enzyme promoter by about 4-fold. In contrast, expression of exogenous RXR alpha in CEH had no effect on the regulation of malic enzyme transcription by T3. These observations support the hypothesis that alterations in RXR alpha expression contribute to cell-type dependent differences in T3 responsiveness of the malic enzyme gene. PMID- 11026557 TI - Differential regulation by FSH and IGF-I of extracellular matrix IGFBP-5 in bovine granulosa cells: effect of association with the oocyte. AB - Inhibition of insulin-like growth factor (IGF)-I induced DNA synthesis in bovine oocyte-cumulus complexes (OCCs) caused by follicle-stimulating hormone (FSH) has been linked to changes in the extracellular matrix which do not occur in mural granulosa cells (MGCs). We investigated regulation by IGF-I and FSH of secreted and extracellular matrix entrapped IGF-binding proteins. OCCs and MGCs from bovine ovaries were cultured in media supplemented with IGF-I and FSH for 24 h. Culture media and extracellular matrix were analysed for IGF-binding proteins by Western ligand blot and immunoblot and found to contain principally IGFBP-3 and 5. The combined treatment of IGF-I and FSH increased the concentration of IGFBP-3 in OCC and MGC conditioned media by 4- and 6-fold, respectively. Treatment of OCCs and not MGCs with IGF-I and FSH together increased extracellular matrix IGFBP-5 by 2.5-fold. The differential regulation of extracellular matrix IGFBP-5 in OCCs compared to MGCs suggest involvement of changes in the extracellular matrix brought about by IGF-I and FSH in overall regulation of IGF-I in the ovarian follicle. PMID- 11026558 TI - Stress-induced mitogen-activated protein kinase signaling in the corpus luteum. AB - Current evidence suggests that stress-induced apoptosis is mediated through the activation of the mitogen-activated protein kinase (MAPK) signaling cascade. We hypothesize that stress-related signaling events documented in other cell lines may also occur in the corpus luteum. To test this, cultured bovine luteal cells were exposed to UV irradiation and harvested at different intervals (0, 30, 120, 240 and 360 min) for analysis of protein or apoptotic cell death. In response to UV treatment cellular levels of phosphorylated p38MAPK and jun-n-terminal kinase (JNK) were increased within 30 min and remained elevated over controls for the duration of the experiment. In contrast, the levels of the phosphorylated forms of p42MAPK and p44MAPK were dramatically reduced. The changes in MAPK signaling were similar to those observed in response to tumor necrosis factor alpha, a cytokine implicated in luteal regression. The UV-induced changes in MAPK phosphorylation were associated with an increase in caspase 3 activity and apoptotic cell death. Taken together, these data demonstrate that stress-induced signaling events in the corpus luteum are similar to those observed in unrelated cell types. Thus, stress-related signaling events may play a role in luteal regression. PMID- 11026559 TI - Immunohistochemical distribution of chicken ovalbumin upstream promoter transcription factor II in human tissues. AB - Chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is an orphan member of the steroid/thyroid hormone receptor superfamily. The present study examines the immunohistochemical distribution of COUP-TFII in human adult tissues. In the reproductive and endocrine systems, COUP-TFII immunoreactivity was detected in the stroma, vascular endothelium and smooth muscle, while it was less frequent in adrenal 4 binding protein (Ad4BP) positive steroidogenic cells. In lung, COUP-TFII immunoreactivity was detected in the vascular endothelium of alveolar septae. In kidney, the glomerular endothelium and Bowman's capsule were immunopositive for COUP-TFII. COUP-TFII immunoreactivity in the gastro-intestinal tract, liver and spleen were detected in mesenchymal cells, sinusoid endothelium and reticuloendothelium, respectively. Results from this study demonstrated the detection of COUP-TFII immunoreactivity in all human tissues examined, especially in mesenchymal cells. The widespread expression of COUP-TFII suggests that COUP TFII may play an important role in the function and homeostasis of various human tissues and organs. PMID- 11026560 TI - Prostaglandin A1 enhances c-fos expression and activating protein-1 activity. AB - Prostaglandin A1 (PGA1) increases heat shock element (HSE)-mediated transcription, thereby enhancing expression of HSE-bearing genes, including heat shock proteins. Because we recently found functional HSEs in the human and rodent c-fos promoters, we hypothesized that PGA1 might increase c-fos expression through the HSE. In this study, we revealed that PGA1 induces c-fos expression at least partly by increasing the binding between heat shock factor-1 and the HSE, and that PGA1 enhances activity of activating protein-1 (AP-1). Interestingly, so far as PGA, is present in the medium, AP-1-mediated transcription enhanced by PGA1 cannot be detected by the standard luciferase reporter gene assay. Instead, it can be detected by either checking luciferase mRNA levels in the presence of PGA1 or measuring luciferase activities just after removal of PGA1. These results showed that protein products of some stress-responsive genes can increase, not during the stressful condition, but immediately after recovery from stress. PMID- 11026561 TI - Alpha2-HSG, a specific inhibitor of insulin receptor autophosphorylation, interacts with the insulin receptor. AB - Human fetuin, [alpha2-Heremans Schmid Glycoprotein (alpha2-HSG)], is a natural inhibitor of insulin receptor tyrosine kinase activity (IR-TKA). Previously, we have demonstrated that alpha2-HSG inhibits the mitogenic pathway without affecting the metabolic arm of insulin signal transduction. In this study, we demonstrate the time-course and specificity of inhibition, its interaction with IR and probable physiological role. In intact rat1 fibroblasts overexpressing the human insulin receptor (HIRc B), incubation of recombinant human alpha2-HSGbac (1.8 microM) inhibited insulin-induced IR autophosphorylation by over 80%. This inhibitory effect of alpha2-HSGbac on insulin-induced IR autophosphorylation was blunted by half in 60 min. Interestingly, alpha2-HSGbac at similar concentrations (0.9 or 1.8 microM), had no effect on EGF- or IGF-I-induced cognate receptor autophosphorylation. Anti-alpha2-HSG immunoprecipitates of alpha2-HSGbac-treated HIRc B cell lysates demonstrated the presence of IR. Our data suggest that alpha2 HSGbac preferentially interacts with the activated IR. To further characterize the site(s) of interaction, the effect of alpha2-HSGbac on trypsin-treated IR autophosphorylation was studied. Trypsin-treatment of intact HIRc B cells results in proteolysis of the IR alpha-chain and constitutive activation of IR-TKA. We demonstrate that alpha2-HSGbac (0.1 microM) completely inhibited trypsin activated IR autophosphorylation and TKA in vitro indicating that this effect was not mediated by its interaction with the proximal 576 amino acid residues of the IR alpha-subunit. The physiological relevance of these observations was explored by characterizing the effects of alpha2-HSG injection in rats. Alpha2-HSGbac (2 microM), acutely injected through the portal vein of normal rats, inhibited insulin-stimulated IR autophosphorylation and IRS-1 phosphorylation in liver and hindlimb muscle. Taken together our results suggest that alpha2-HSG, by interacting with IR, specifically inhibits insulin-stimulated IR autophosphorylation and may play a physiological role in the regulation of insulin signaling. PMID- 11026562 TI - Transcription factor expression during pancreatic islet regeneration. AB - Recent studies by a number of laboratories have identified transcription factors that are involved in pancreatic development. Indeed, marked abnormalities in pancreatic development result from deficiencies in these molecules, which include, among others, PDX-1, islet-1 (Isl-1), and Pax-6. These studies have prompted us to evaluate the expression of Isl-1 and Pax-6 in the pancreas of the interferon-gamma (IFNgamma) transgenic mouse, which exhibits new islet growth and expansion of ducts throughout the life of the animal. We have previously demonstrated that PDX-1 is strikingly expressed in the ducts of the IFNgamma transgenic mouse. This latter observation compelled us to examine expression of hepatocyte nuclear factor-3beta (HNF3beta), which mediates PDX-1 gene transcription, in the IFNgamma transgenic pancreas as well. As a result of these studies, we now demonstrate marked expression of these transcription factors in the pancreatic ducts of IFNgamma transgenic mice. These data suggest a role for these transcription factors during pancreatic regeneration in the IFNgamma transgenic mouse. PMID- 11026563 TI - Differential expression of estrogen receptors alpha and beta in adult rat accessory sex glands and lower urinary tract. AB - Estrogens induce pronounced structural and functional changes in male accessory sex glands and the lower urinary tract in both sexes, but the exact mechanisms of estrogen action are not fully understood. This study was undertaken to localise the tissue cell types that express estrogen receptor in adult rats, and to determine the receptor subtype (ER alpha and ER beta) in order to identify sites that may respond directly to estrogens. In the male accessory sex glands (seminal vesicles, prostatic lobes and ampullary glands), ER beta mRNA and protein were strongly expressed in the epithelium but not in the stroma, while ER alpha mRNA was present only in the fibromuscular tissue surrounding the prostatic collecting ducts in the posterior periurethral region and in ampullary gland stroma. In the epithelium of the urinary bladder and urethra of both sexes, high level of ER beta mRNA and protein, but no ER alpha mRNA, was detected. The connective tissue in urinary bladder of both males and females, as well as that in prostatic urethra in males expressed ER alpha mRNA. The neural cells in the autonomic ganglia of the prostatic plexus were strongly positive for ER beta mRNA, but were completely devoid of ER alpha. We conclude that ER beta is the predominant ER subtype in the epithelium of adult male rat accessory sex glands and the lower urinary tract of both males and females, as well as in the prostatic neural plexus regulating the function of the lower urinary tract in males, while ER alpha is present only in the stromal compartment of distinct sites. These results indicate that in these tissues in intact adults there are multiple targets for direct estrogen action. Furthermore, the differential or complementary expression of the two ER subtypes suggests that they may have specific functions, and may explain the complex structural and functional changes induced by estrogens. PMID- 11026564 TI - Induction of progesterone receptor immunoexpression in stromal tissue throughout the male reproductive tract after neonatal oestrogen treatment of rats. AB - Oestrogen exposure of the male during fetal/neonatal life can fundamentally alter the structure and function of the reproductive system, though how is unknown. This study examined whether such treatment was able to induce a 'female' characteristic, namely immunoexpression of progesterone receptor (PR), in the reproductive system of the male. Rats were treated on postnatal days 2, 4, 6, 8, 10 and 12 with either 10, 1 or 0.1 microg diethystilbestrol (DES) or with the vehicle (20 microl corn oil). Groups of control and treated rats were killed on days 18, 25, 35 and 90 (= adults) and tissues fixed in Bouins for immunolocalisation studies using antisera to PR (recognises A and B forms) and oestrogen receptor-beta (ER beta). PR immunoexpression was absent from all tissues studied in control rats at all ages with the exception of the parasympathetic ganglia of the prostate. In rats treated with 10 microg DES, intense immunoexpression of PR was detected in the nuclei of stromal, but not epithelial, cells of the caput and cauda epididymis, the vas deferens, seminal vesicles and at the base of the dorsolateral prostatic complex (DLPC) at day 18, but was absent from the ventral prostate and from the testis. DES induction of PR immunoexpression was evident after a single injection (on day 3) and at 18-35 days the intensity of immunoexpression was DES dose-dependent; rats treated neonatally with 0.1 microg DES showed no detectable PR immunoexpression at any age. These findings were confirmed by Western analysis which indicated that most of the PR induced was probably the B form. Co-localisation studies, using confocal microscopy, demonstrated that PR and ER beta frequently co-localised to the same stromal cells in the DLPC, epididymis and seminal vesicles of DES treated rats at day 18, whereas epithelial cells, which also expressed ER beta, did not express PR. In the tissues studied, only occasional stromal cells expressed ER alpha in comparison to the more widespread expression of ER beta, although epithelial cell expression of ER alpha was also detected in the epididymis on day 18 (but not on day 10). In DES-treated rats, immunoexpression of PR in the reproductive tract decreased progressively in intensity from days 18 35 and was non-detectable in adulthood. In conclusion, these findings are interpreted as evidence that neonatal oestrogen treatment exerts pervasive 'reprogramming' effects throughout the reproductive system of the developing male as indicated by the induction of PR immunoexpression. This induction was restricted to stromal tissue even though both stromal and epithelial cells at most sites expressed ER beta and/or ER alpha. PMID- 11026565 TI - 1,25-Dihydroxyvitamin D3 decreases human prostate cancer cell adhesion and migration. AB - 1,25-Dihydroxyvitamin-D3 [1,25(OH)2D3], the active hormonal metabolite of vitamin D, acts through a specific nuclear receptor to inhibit proliferation and promote differentiation of several tumor cell types including the LNCaP, DU145 and PC-3 prostate cancer cell lines as well as primary prostate tumor lines. 1,25(OH)2D3 can also decrease invasion of breast and prostate cancer cell lines in vitro. We confirm this latter finding in the DU145 and PC-3 prostate cancer cell lines, and further show that 1,25(OH)2D3 inhibits overall invasion, cell adhesion and migration to the basement membrane matrix protein laminin. These changes appear to be due in part to a 1,25(OH)2D3-induced decrease in expression of alpha6 and beta4 integrins, both of which are receptors for laminin and associated with increased migration and invasion of prostate cancer cells in vitro. Blocking function of these particular integrins with antibodies inhibits both adhesion and migration of the cells. Collectively, these data demonstrate that 1,25(OH)2D3, in addition to decreasing proliferation of tumor cells, can also inhibit prostate cancer cell invasion through modulation of select cell surface adhesion molecules. PMID- 11026566 TI - The role of insulin dissociation from its endosomal receptor in insulin degradation. AB - Mechanisms that terminate signals from activated receptors have potential to influence the magnitude and nature of cellular responses to insulin. The aims of this study were to determine in rat liver endosomes (the subcellular site of insulin signal termination) whether dissociation of insulin from its receptor was a pre-requisite for ligand degradation and whether the state of receptor phosphorylation influenced the dissociation and hence endosomal degradation of insulin and/or receptor recycling. Following in vivo administration of 125I-[A14] insulin or analogues (native, X10 or H2, relative binding affinities 1:7:67) livers were removed and endosomes prepared. In the endosomal preparations a significantly greater percentage of both analogues were receptor-bound than native insulin with concomitantly less ligand degradation. When rats were injected with protein-tyrosine phosphatase inhibitors (peroxovanadium compounds bpV(phen) or bpV(pic)) before insulin, endosomal insulin receptor phosphotyrosine content, assessed by Western blotting, was increased as was receptor-bound 125I [A14]-insulin, whilst insulin degradation was decreased. Peroxovanadiums also completely inhibited recycling of insulin receptors from endosomes. However, treatment of freshly isolated endosomes with acid phosphatase which completely dephosphorylated the insulin receptor, did not return the rate of insulin dissociation and degradation to control values, suggesting that peroxovanadium compounds elicit their effect on binding and degradation via a mechanism other than as protein-tyrosine phosphatase inhibitors. We conclude that promotion of sustained receptor binding decreases endosomal insulin degradation and extends the half-life of the activated endosomal receptor, which in turn would be expected to potentiate insulin signalling from this intracellular compartment. PMID- 11026568 TI - Characterization of gonadal and extra-gonadal forms of the cDNA encoding the Atlantic stingray (Dasyatis sabina) cytochrome P450 aromatase (CYP19). AB - Cytochrome P450 aromatase (P450arom; CYP19) mediates the conversion of androgens to estrogens and its activity has been found in all vertebrates studied to date. This study describes the full-length cDNA encoding the ovarian form of P450arom and the differences in the 5'-untranslated region (5'-UTR) of the extra-gonadal P450arom transcript expressed by the Atlantic stingray (Dasyatis sabina). Elasmobranchs (cartilaginous fishes such as sharks, rays and skates) diverged from the other vertebrates more than 350 million years ago, therefore the stingray P450arom cDNA may represent an ancient form of this gene. Northern blot analysis showed that the ovarian follicle expressed transcripts of 3.1 and 1.7 kb in size which correspond to the clones isolated from a stingray ovarian follicle cDNA library. Both transcripts consisted of an identical 1.5 kb coding region and a 41 bp 5'-UTR, however the 3'-UTRs differed in the use of the most proximate and the most distal of four polyadenylation signals. COS cells transfected with the 1.7 kb cDNA had twice the aromatase activity as cells transfected with the 3.1 kb cDNA. The coding region of the cDNA predicted a 58.5 kDa protein which consisted of 511 residues. Alignment of the stingray protein indicates that the P450arom is equally identical (53-59%) to all other vertebrate forms of P450arom characterized to date, thus indicating a common ancestry. The evolutionary relationship of the stingray form of P450arom clearly predates the other forms and belongs to a unique lineage. Transcripts of P450arom were expressed in ovarian follicles (of all sizes), the testis, the pituitary, in all sections of the brain, and in the kidney. The extra-gonadal transcripts appear to encode a protein identical to the ovarian form, however, the 5'-UTR was 657 bp longer presumably due to the transcription of an untranslated 'first exon' as seen in the mammalian form of this gene. PMID- 11026567 TI - The paradoxical effect of adrenomedullin on Na+ transport by the renal distal tubule luminal membrane. AB - Adrenomedullin (ADM) is a potent hypotensive and natriuretic peptide which is synthetized in several mammalian tissues including the kidney. The purpose of this study was to investigate whether the natriuresis was due to a change in Na+ transport by either the proximal (PT) or the distal tubule (DT) luminal membrane, and to characterize this effect, if present. PT and DT suspensions were incubated with human ADM for 20 min at 37 degrees C and luminal membranes of these tubules were purified using the Mg2+ precipitation technique. Na+ uptake was measured by the Millipore filtration technique. A volume of 10(-8) M ADM had no effect on Na+ uptake by the PT luminal membranes. In contrast and unexpectedly, the hormone increased Na+ transport by the DT membranes from 0.28 +/- 0.03 to 0.68 +/- 0.06 pmol/microg per 5 s (P < 0.01). The dose-response curve of this effect showed a maximal response with 10(-7) M ADM. The hormone influenced exclusively the Na+/H+ exchanger, leaving the N-ethyl-N-isopropyl-amiloride (EIPA) insensitive transport intact. The addition of Rp cAMPs to the preparations completely abolished the effect of the hormone on Na+ transport suggesting that cAMP was the messenger involved in this action. Finally, incubation of the DT suspensions with aldosterone also stimulated 1 mM Na+ uptake by the luminal membrane and the two hormone actions were not additive. We conclude that, although ADM is a natriuretic hormone probably through its vasodilating action, it increases distal Na+ reabsorption by the stimulation of the Na+/H+ exchanger activity, as does aldosterone at the same site. PMID- 11026569 TI - Isolation of differentially expressed aldose reductase in ovaries after estrogen withdrawal from hypophysectomized diethylstilbestrol treated rats: increased expression during apoptosis. AB - More than 99% of the follicles are eliminated by apoptosis before reaching ovulation. Several growth factors and hormones inhibit apoptosis in the ovary, including estrogen. Using differential display of mRNA, aldose reductase was shown to increase in the ovary of diethylstilbestrol treated hypophysectomized rats after estrogen withdrawal, inducing apoptosis. The aldose reductase mRNA expression was confirmed to be 2.2 +/- 0.2-fold higher after estrogen withdrawal using northern blot analysis. In addition, untreated immature rats showed a 1.7 +/- 0.3-fold higher expression of ovarian aldose reductase mRNA compared to ovaries 24 h after pregnant mare's serum gonadotropin treatment, decreasing apoptosis in the ovary. In the prostate, the level of aldose reductase was increased 3.1 +/- 1.1-fold 2 days after castration induced apoptosis. Although the physiological role of aldose reductase in the ovary is not known, these data suggest that aldose reductase may be part of a hormonally regulated apoptotic pathway in the ovary and prostate. PMID- 11026570 TI - Effects of amphotericin B and ketoconazole on mouse oocyte maturation: implications on the role of meiosis-activating sterol. AB - Meiosis-activating sterol (MAS) has been shown to induce mouse oocytes cultured in the presence of hypoxanthine (HX) to resume meiosis. The present research was conducted to determine whether amphotericin B or ketoconazole (a promoter and an inhibitor of production of MAS), affected oocyte maturation. Mouse cumulus cell enclosed oocytes (CEO) or denuded oocytes (DO) were cultured for 24 h in the presence of 4 mM HX with FSH or amphotericin B or ketoconazole. At the end of the culture, the frequency of germinal vesicle break down (GVBD) and polar body formation (PB) were recorded. The results demonstrated: (i) FSH (10-200 IU/l) induced dose-dependent oocytes maturation in CEO, but was without effect on DO. A maximum increase in GVBD and PB was observed with 25-50 IU/l FSH. The presence of FSH (50 IU/l) for 1 h was sufficient to induce meiotic resumption, which after 2 h reached a plateau similar to that of a continuous presence of FSH. (ii) CEO exposed to amphotericin B (0.0025-2.5 microg/l) underwent GVBD dose-dependently, whereas no effect was observed on DO. The presence of amphotericin B (0.025 microg/l) for 1 h stimulated oocyte resumption in a way similar to that of FSH. (iii) Amphotericin B (0.025 microg/l) and FSH (50 IU/l) did not show any additive effect on resumption of meiosis. (iv) Ketoconazole (10(-7)-10(-3) M) inhibited the effect of FSH on resumption of meiosis, but had no effect on oocyte spontaneous maturation. These results show that FSH and amphotericin B induce resumption of meiosis and indicate that they are likely to cause an accumulation of meiosis activating sterols in the CEO, but ketoconazole blocks the production of MAS. The present study supports the notion that MAS plays a physiological relevant role in triggering resumption of meiosis in mouse oocytes. PMID- 11026571 TI - Molecular cloning, distribution and pharmacological characterization of a novel gonadotropin-releasing hormone ([Trp8] GnRH) in frog brain. AB - To date nine structural variants of GnRH have been identified in vertebrates and two additional forms have been isolated from a tunicate. In amphibians only mammalian GnRH ([Arg8] GnRH) and type II GnRH (chicken GnRH II, [His5, Trp7, Tyr8] GnRH) have been identified. In the present study, a full-length cDNA encoding a novel type of GnRH was isolated from pituitary of Rana dybowskii. The GnRH gene encodes a GnRH peptide ([Trp8] GnRH) in which tryptophan is substituted for arginine of mammalian GnRH Northern blot analysis revealed the presence of a single 500 bp transcript for the [Trp8] GnRH precursor in forebrain but its absence in testis, ovary, kidney and liver. Restriction digests of genomic DNA demonstrated a single copy of the gene. The [Trp8] GnRH immunoreactive cells were identified in the preoptic area of the frog brain. Synthetic [Trp8] GnRH was tested for its ability to stimulate inositol phosphate production by COS-1 cells transfected with the cloned Xenopus pituitary GnRH receptor and the cloned human GnRH receptor. [Trp8] GnRH had a potency of about 60% compared with mammalian GnRH ([Arg8] GnRH) for the Xenopus receptor, whereas the potency of [Trp8] GnRH was approximately 5% compared with mammalian GnRH for the human receptor. Both mammalian GnRH and [Trp8] GnRH were 1000-fold less potent than type II GnRH for the Xenopus GnRH receptor. The similar potency of [Arg8] GnRH and the novel [Trp8] GnRH for the Xenopus pituitary receptor indicates that, unlike the human receptor, the Xenopus receptor does not discriminate between these amino acids in position eight thereby allowing substitution of the arginine in the mammalian GnRH. PMID- 11026572 TI - Involvement of Sp1 in the transcriptional regulation of the rat insulin-like growth factor-1 gene. AB - Most insulin-like growth factor-I (IGF-I) transcripts are initiated in exon 1, but mechanisms of regulation are not well understood. Since potential Sp1 sites are found in footprinted regions within approximately 360 bp upstream and downstream from the major initiation sites in exon 1, we explored the involvement of Sp1 and Sp3 in regulation of IGF-1 expression. Gel shift assays showed strong Sp1 binding to the downstream site, but binding to the upstream site was weak; Sp1 bound to a CCTGCCCA sequence in downstream footprint region V, and Sp3 binding was centered on the same sequence. IGF-I basal promoter constructs containing a mutation in the downstream Sp1 site exhibited a 32% decrease in expression in CHO cells and a 75% decrease in HepG2 cells, indicating the importance of Sp1 for expression in vivo. Sp1 and Sp3 expression vectors provided three- to five-fold stimulation of wild-type IGF-I constructs, but had little effect on a construct containing a mutation in the downstream Sp1 site, and Sp1 had comparable effects in Drosophila SL2 cells. IGF-I heterologous promoter constructs exhibited similar responses: in both SL2 cells and CHO cells, stimulation by Sp1 was enhanced with constructs containing downstream region V. Since Sp1 also stimulated expression of concatamers of putative cis-acting sites fused to the SV40 promoter enhancer in pGL3, the results in combination indicate that the presence of IGF-I region V is sufficient to permit stimulation by Sp1. CONCLUSION: Sp1 and related factors may play an important role in the regulation of IGF-I gene transcription, through interactions with region V downstream from the major initiation sites in exon 1. PMID- 11026573 TI - High level, tissue-specific expression of a modified calcitonin/calcitonin gene related peptide promoter in a human medullary thyroid carcinoma cell line. AB - The efficient and high level expression of therapeutic genes in target cells is critical for effective gene therapy. We have developed a novel promoter by utilizing tandem repeats of a tissue-specific regulatory element from the calcitonin/calcitonin gene-related peptide (CT/CGRP) gene placed in close proximity to a basal promoter, thereby removing interstitial sequences. This promoter drives expression of reporter genes at much higher levels than the natural promoter while significantly improving specificity in thyroid C cells. PMID- 11026574 TI - Regulation of keratin 19 gene expression by estrogen in human breast cancer cells and identification of the estrogen responsive gene region. AB - Estrogens regulate the proliferation, cytoarchitectural, and invasive properties of estrogen receptor (ER)-containing breast cancer cells. To identify genes under direct regulation by estrogen in breast cancer cells, we have used representational difference analysis (RDA) of cDNAs. In this way, we have identified (cyto)keratin 19 (K19), a major component of cell intermediate filaments, as being under rapid and direct regulation by estrogen in MCF-7 cells. Stimulation by estradiol (E2) of K19 mRNA is rapid, with maximal increase at 3 h, and is not blocked by cycloheximide, suggesting that it is a primary response to the hormone. Increased accumulation of K19 protein is observable by 8 h after E2 and levels continue to increase at 24-48 h after E2 treatment. Suppression of E2 induced K19 gene expression by the antiestrogen ICI 182,780 suggests that ER mediates this regulation. Analysis of the human K19 chromosomal gene, by transient transfection assays employing reporter gene constructs with the 5' and 3' flanking regions and portions of the body of the K19 gene, has resulted in identification of a complex enhancer region in the first intron. This enhancer region consists of a near-consensus estrogen response element (K19 ERE, which differs by only 1 bp from the consensus ERE) and two ERE half sites, as well as two AP1-like sites. The results of transfections with either the K19 gene promoter or the heterologous thymidine kinase promoter and constructs containing mutated or deleted portions of the enhancer region show that the K19 ERE is responsible for the E2-dependent transactivation of the keratin 19 gene and for the synergism that is observed between E2 and TPA with both ER alpha and ER beta. These studies document ER regulation of the K19 gene, localize the estrogen responsive region, and suggest that up-regulation of keratin 19 gene expression by estrogen may contribute to the cytoskeletal and nuclear matrix reorganization, and increased metastatic potential of ER-containing breast cancer cells upon exposure to estrogens. PMID- 11026576 TI - Intrauterine growth restriction and Doppler ultrasonography. AB - To clarify the difference between the fetus that is small for gestational age and the fetus with true intrauterine growth restriction, we undertook a retrospective study of singleton fetuses who had fetal weight estimation and umbilical artery Doppler velocity studies within 2 weeks of their delivery. Fetuses were divided into four categories on the basis of sonographic results from their last examination. Statistical comparisons of neonatal outcome were made for the four groups, which totaled 578 fetuses. Increased cesarean section for fetal distress, stays in the neonatal intensive care unit, and increased neonatal morbidity were seen in both small for gestational age and average for gestational age neonates with abnormal Doppler blood flow. The small for gestational age fetuses with normal Doppler studies showed no increased morbidity when compared with their average for gestational age cohorts. Umbilical artery Doppler blood flow studies were a better predictor of neonatal outcome than estimated fetal weight. Small for gestational age fetuses with normal Doppler studies most likely represent constitutionally small, not pathologically growth restricted, fetuses. PMID- 11026575 TI - Upregulation of pancreatic polypeptide-sensitive neuropeptide Y (NPY) receptors in estrogen-induced hypertrophy of the anterior pituitary gland in the Fischer 344 rat. AB - Implants of diethylstilbestrol inducing anterior pituitary prolactinomas in female Fischer-344 rats produced a considerable elevation of high-affinity binding of either rat or human pancreatic polypeptide (hPP). No comparable upregulation of high-affinity binding sites was noted for the ligand [125I](Leu31,Pro34)hPYY (LP-PYY) (masked by 2 nM hPP to force selectivity for the Y1 sites), or of the Y2-selective ligand [125I]hPYY(3-36). The Y5-like sites displayed the rank order of potency of hPP = rPP, hNPY, LP-PYY > pPYY(1-36) > hNPY(2-36) > hPYY(3-36) >> aPP, similar to the previously described rabbit kidney or hypothalamic Y5-like receptors. The PP binding in the anterior pituitary was not sensitive to the Y1-selective non-peptidic antagonist BIBP-3226. The PP binding was highly sensitive to alkali metal cations, and to a N5-substituted amiloride antagonist of sodium transport, but not to a C2-guanido substituted amiloride antagonist of sodium channels. The binding was also sensitive to phospholipase C antagonist U-73122, and to alkylating alpha-adrenergic agonist chloroethylclonidine. Lactotrophs isolated in Percoll gradients after enzymic dispersion of the anterior pituitary gland retained the high-affinity PP binding. Following removal of estrogen implants, the hPP binding sites decreased to very low levels within 3 days, in parallel to the decrease of plasma prolactin. PMID- 11026577 TI - Doing away with the full bladder in doing pelvic ultrasonography (transabdominally). PMID- 11026578 TI - Fetal cardiac dextroposition in the absence of an intrathoracic mass: sign of significant right lung hypoplasia. AB - We reviewed our experience of fetal cardiac dextroposition in the absence of an intrathoracic mass. Ten cases were found by fetal echocardiography to have a normal cardiac axis, but the heart was shifted into the right chest and the amount of right lung tissue was reduced. At birth seven of the infants had confirmed structural heart disease (70%), including three with scimitar syndrome. Two infants had additional extracardiac anomalies (20%). Seven infants born at term had clinical pulmonary hypertension with a diagnosis of right lung hypoplasia in all of them. Two neonates died owing to significant heart disease (one with scimitar syndrome and the other with hypoplastic left heart syndrome). Of the three pregnancies that were terminated, the two fetuses with autopsies had severe right lung hypoplasia. Fetal cardiac dextroposition and right pulmonary artery hypoplasia in the absence of an intrathoracic mass are important signs of right lung hypoplasia, which can be associated with significant pathologic cardiac and extracardiac conditions. PMID- 11026579 TI - Ultrasonographic tissue characterization in monitoring tumor response to neoadjuvant chemotherapy in locally advanced breast cancer (work in progress). AB - A program of computer-assisted texture analysis was applied to evaluate its usefulness for objective description of changes in tumor architecture due to primary medical treatment in patients with locally advanced breast cancer. Changes in values of parameters of the statistical pattern recognition technique were compared to ultrasonographically depictable, subjectively recorded changes in echogenicity and echotexture (brightness, homogeneity) and reviewed with regard to histopathologic evaluation of tumor regression. Characteristic trends of defined quantitative texture parameters (mean gradient, mean gray value, contrast from the co-occurrence matrix) corresponded to visually depictable changes of the B-mode image and underlying histopathologic changes. The results indicate that quantitative texture analysis may aid in noninvasive monitoring of tumor response to neoadjuvant chemotherapy. PMID- 11026580 TI - The correlation between histologic features and the echogenicity in Asian hepatocellular carcinomas (HCCs) is not applicable to non-Asian HCCs. PMID- 11026581 TI - Efficacy of community-based second trimester genetic ultrasonography in detecting the chromosomally abnormal fetus. AB - We sought to assess prospectively the efficacy of community-based genetic ultrasonography in detecting chromosomally abnormal fetuses in a high-risk population and determine independent markers of aneuploidy. Patients 18 years old and older who were between 14 and 24 weeks' gestation were included if referred for maternal age greater than 35 years, increased risk of Down syndrome or trisomy 18 by second trimester serum screen, or prior affected offspring. All women had a targeted ultrasonographic examination between April 1997 and June 1999 and were offered fetal chromosomal analysis. Markers of aneuploidy and pregnancy outcomes were recorded prospectively. The primary outcome was prenatally or postnatally detected chromosomal abnormalities. Of the 1030 fetuses seen during the study, 789 had outcome data available and constituted the study group. In this group, 694 (87.9%) ultrasonograms were normal, 73 (9.2%) had one marker present, 17 (2.2%) had two markers present, and 5 (0.6%) had three or more markers present. Fourteen of 17 (82.3%) aneuploid fetuses had an abnormal ultrasonogram (one or more markers present), including 5 of 7 (71.4%) with Down syndrome. Logistic regression showed abnormal four-chamber view, structural anomaly, and intracardiac echogenic focus to be significant aneuploidy markers. The amniocentesis rate was 334 of 1030 (32.4%), and it increased with the number of sonographic markers noted (0 = 29.9%, 1 = 60.2%, 2 = 70.6%, 3 or more = 80%). Genetic ultrasonography is highly effective in identifying chromosomally abnormal fetuses in a community-based practice. PMID- 11026582 TI - Role of color Doppler ultrasonography in the diagnosis of endometriotic cyst. AB - We studied the role of color Doppler ultrasonography in the distinction between endometriomas and other adnexal masses. Three hundred and fifty-two ovarian lesions were studied, comparing sonographic diagnosis with pathologic findings. On color Doppler sonography, an endometriotic cyst usually appeared as a cystic lesion with diffuse internal echoes and low vascularization. The sensitivity and specificity of color Doppler transvaginal sonography in detecting endometriotic cysts were 91.8% and 95.3%, respectively. The positive and negative predictive values were 95.5% and 91.5%, respectively. In our experience, transvaginal sonography with color Doppler interrogation is a useful technique in the diagnosis of pathologic ovarian conditions, including cystic endometriosis. PMID- 11026583 TI - Three-dimensional color Doppler sonography before and after fibroid embolization. AB - Three-dimensional color Doppler sonography was performed within 1 hour, 1 day, 3 months, and 6 months of fibroid embolization in 20 patients who had a total of 31 fibroids greater than 2 cm in average dimension. The greatest decrease in vascularity occurred 1 day after the procedure, whereas the greatest volume change was found at the 3 month follow-up examination. In about one half of the patients scanned, depiction of fibroid vascularity by color Doppler sonography was found to improve the delineation of the size, location, and extent of myometrial involvement. Hypervascular fibroids (12 of 31) tended to decrease in size after treatment more than isovascular (10 of 31) or hypovascular ones (9 of 31). Additional investigations that are similar to this one will be needed to determine if three-dimensional color Doppler sonography can be used to predict those who will be responders, partial responders, or nonresponders to embolotherapy. PMID- 11026584 TI - Focal adherent thrombus in the common carotid artery: clinical, ultrasonographic, and pathogenic aspects in two cases. PMID- 11026585 TI - Adventitial cystic disease of the popliteal artery demonstrated by intravascular ultrasonography. PMID- 11026586 TI - Chondrodysplasia punctata, tibial-metacarpal type in a 16 week fetus. PMID- 11026587 TI - Prenatal ultrasonographic diagnosis of ileal atresia and volvulus in a twin pregnancy. PMID- 11026588 TI - UNWIRED E-MED: the next generation of wireless and internet telemedicine systems. AB - The movement of telemedicine to wireless and mobile Internet applications is imminent in the next few years. This migration from desktop platforms to wireless and mobile configurations will have a significant impact on future healthcare delivery systems and their globalization. Recent telecommunications and biomedical computing advances will significantly enhance the current methodologies of telemedicine and telecare systems. This editorial will present some of the evolutionary issues and important aspects that have to be considered in developing technologies for the next generation of Internet and third generation of mobile systems geared for future telemedical applications. These will provide new dimensions to existing medical services and areas of outreach that are not possible with the current generation that will have tremendous impact on how healthcare delivery will be shaped for the 21st century. PMID- 11026589 TI - Construction of a regional telementoring network for endoscopic surgery in Japan. AB - This paper introduces a regional telementoring network for endoscopic surgery in Japan. The telecommunication center was established at Osaka University and it has been operational since 1997 using six ISDN B-channels. The network, composed of five remote hospitals, aimed to tele-educate young surgeons in constituent hospitals, and it was applied not only to tele-education in routine endoscopic surgery, but also to telementoring in advanced operations. As the significance of telementoring in surgery has been gradually recognized, several networks following our model are now under consideration in other areas of Japan. Although there remains several problems around wide and general propagation of telesurgery, such as coverage of the expenditure or privacy protection, it will be beneficial to prevent professional isolation to share limited resources of surgical experts, and it is expected to contribute to patients receiving advanced surgical therapies. PMID- 11026590 TI - Optimal zonal wavelet-based ECG data compression for a mobile telecardiology system. AB - A new integrated design approach for an optimal zonal wavelet-based ECG data compression (OZWC) method for a mobile telecardiology model is presented. The hybrid implementation issues of this wavelet method with a GSM-based mobile telecardiology system are also introduced. The performance of the mobile system with compressed ECG data segments selected from the MIT-BIH arrhythmia data base is evaluated in terms of bit error rate (BER), percent rms difference (PRD), and visual clinical inspection. The compression performance analysis of the OZWC is compared with another wavelet-based (Discrete Symmetric Wavelet Compression) approach. The optimal wavelet algorithm achieved a maximum compression ratio of 18: 1 with low PRD ratios. The mobile telemedical simulation results show the successful compressed ECG transmission at speeds of 100 (km/h) with BER rates of less than 10(-15), providing a 73% reduction in total mobile transmission time with clinically acceptable reconstruction of the received signals. This approach will provide a framework for the design and functionality issues of GSM-based wireless telemedicine systems with wavelet compression techniques and their future integration for the next generation of mobile telecardiology systems. PMID- 11026591 TI - A WEB-based telePACS using an asymmetric satellite system. AB - We have developed a WWW-based TelePACS that can access every permitted PACS server via the Internet. Java programming techniques were used to implement the system, which can access and retrieve medical information and images through Web browsers only such as Netscape without specific tools. We also have developed a consolidator that performs as a manager to connect a conventional PACS server to a Web-based TelePACS server. We have developed the Asymmetric Satellite Data Communication System (ASDCS) as a fast communication system. The ASDCS uses a receive-only satellite link for data delivery and a terrestrial network for control communication. In conclusion, we were able to develop a cost-effective and fast PACS using Web technology. Web technology expanded the scope of use for a dedicated PACS from intrahospital to public use. PMID- 11026592 TI - Detection of multiple sclerosis with visual evoked potentials--an unsupervised computational intelligence system. AB - This paper describes the application of a novel unsupervised pattern recognition system to the classification of the Visual Evoked Potentials (VEP's) of normal and multiple sclerosis (MS) patients. The method combines a traditional statistical feature extractor with a fuzzy clustering method, all implemented in a parallel neural network architecture. The optimization routine, ALOPEX, is used to train the network while decreasing the likelihood of local solutions. The unsupervised system includes a feature extraction and clustering module, trained by the optimization routine ALOPEX. Through maximization of the output variance of each node, and an architecture which excludes redundancy, the feature extraction network retains the most significant Karhunen-Loeve expansion vectors. The clustering module uses a modification to the Fuzzy c-Means (FCM) clustering algorithms, where ALOPEX adjusts a set of cluster centers to minimize an objective error function. The result combines the power of the FCM algorithms with the advantage of a more global solution from ALOPEX. The new pattern recognition system is used to cluster the VEP's of 13 normal and 12 MS subjects. The classification with this technique can, without supervision, separate the patient population into two groups which largely correspond to the MS and control subject groups. A suitable threshold can be chosen so that the recognizer chooses no false negatives. The use of multiple stimulation patterns appears to improve the reliability of the decision. The reasoning of most neural networks in their decision making cannot easily be extracted upon the completion of training. However, due to the linearity of the network nodes, the cluster prototypes of this unsupervised system can be reconstructed to illustrate the reasoning of the system. In this application, this analysis hints at the usefulness of previously unused portions of the VEP in detecting MS. It also indicates a possible use of the system as a training aide. PMID- 11026594 TI - Simulated generation of evoked potentials components using networks with distinct excitatory and inhibitory neurons. AB - Long latency evoked potentials (EP's) are electrical potentials related to brain information processing mechanisms. In this paper, three-layered neurophysiologically based artificial neural network model is presented whose neurons obey to Dale's law. The first two layers of the network can memorize and recall sparsely coded patterns, oscillating at biologically plausible frequencies. Excitatory low-pass filtering synapses, from the second to the third layer, create evoked current dipoles, when the network retrieves memories related to stimuli. Based on psychophysiological indications, simulated intracranial dipoles are straightforwardly transformed into long latency EP components such as N100 and P300 that match laboratory-measured scalp EP's. PMID- 11026593 TI - A family competition evolutionary algorithm for automated docking of flexible ligands to proteins. AB - In this paper, we study an evolutionary algorithm for flexible ligand docking. Based on family competition and adaptive rules, the proposed approach consists of global and local strategies by integrating decreasing mutations and self-adaptive mutations. To demonstrate the robustness of the proposed approach, we apply it to the problems of the first international contests on evolutionary optimization. Following the description of function optimization, our approach is applied to a dihydrofolate reductase enzyme with the anti-cancer drug methotrexate and with two analogs of the antibacterial drug trimethoprim. Our numerical results indicate that the proposed approach is robust. The docked lowest energy structures have rms derivations ranging from 0.72 A to 1.98 A with respect to the corresponding crystal structure. PMID- 11026595 TI - Application of an object-oriented programming paradigm in three-dimensional computer modeling of mechanically active gastrointestinal tissues. AB - The aim of this study was to develop a novel three-dimensional (3-D) object oriented modeling approach incorporating knowledge of the anatomy, electrophysiology, and mechanics of externally stimulated excitable gastrointestinal (GI) tissues and emphasizing the "stimulus-response" principle of extracting the modeling parameters. The modeling method used clusters of class hierarchies representing GI tissues from three perspectives: 1) anatomical; 2) electrophysiological; and 3) mechanical. We elaborated on the first four phases of the object-oriented system development life-cycle: 1) analysis; 2) design; 3) implementation; and 4) testing. Generalized cylinders were used for the implementation of 3-D tissue objects modeling the cecum, the descending colon, and the colonic circular smooth muscle tissue. The model was tested using external neural electrical tissue excitation of the descending colon with virtual implanted electrodes and the stimulating current density distributions over the modeled surfaces were calculated. Finally, the tissue deformations invoked by electrical stimulation were estimated and represented by a mesh-surface visualization technique. PMID- 11026596 TI - An adaptive 3-D discrete cosine transform coder for medical image compression. AB - In this communication, a new three-dimensional (3-D) discrete cosine transform (DCT) coder for medical images is presented. In the proposed method, a segmentation technique based on the local energy magnitude is used to segment subblocks of the image into different energy levels. Then, those subblocks with the same energy level are gathered to form a 3-D cuboid. Finally, 3-D DCT is employed to compress the 3-D cuboid individually. Simulation results show that the reconstructed images achieve a bit rate lower than 0.25 bit per pixel even when the compression ratios are higher than 35. As compared with the results by JPEG and other strategies, it is found that the proposed method achieves better qualities of decoded images than by JPEG and the other strategies. PMID- 11026597 TI - New features of renal lesion induced by stroma free hemoglobin. AB - This study focused on the subacute renal lesions resulting from the infusion of stroma free hemoglobin (SFH), which remains under evaluation as a potential blood substitute despite limited renal toxicity observed in acute infusion. Four groups of rats received different doses of SFH (0.03, 0.48, 0.96, and 1.46 g, respectively) and were monitored, on alternate days, for their glomerular filtration rate over the course of 10 days. Another group of 6 rats receiving 0.96 g SFH was sacrificed at day 10 for examination of renal morphology. The low dose (0.03 g) of SFH infusion did not alter the creatinine clearance (Clcr) over 10 days. The Clcr decreased in rats receiving 0.48 g SFH but fully recovered at day 10. A persistent decrease in Clcr was observed in the groups of rats receiving 0.96 and 1.68 g of SFH. Tubular necrosis was the most prominent renal lesion distributed in the proximal tubules, especially in the convoluted segment of the juxtamedullary nephrons. Pearls' stained cytoplasmic granules and electron dense lysosomal granules were found in surviving proximal tubules. Necrosis was the predominant mechanism of cell death. This study revealed for the first time proliferation of smooth endoplasmic reticulum in the proximal tubules after SFH treatment, where it appeared as nodular aggregates of tubulovesicular structures. The effect of SFH on the proximal tubule appeared to be a direct toxicity, and this toxicity was shown to be dose dependent. The presence of reversible toxicity indicated that a safety limit dosage for SFH infusion exists and that tolerance dose of SFH can be determined for clinical applications. PMID- 11026598 TI - Cecal torsion in rodents associated with chronic administration of clinafloxacin. AB - The chronic toxicity of clinafloxacin, a fluoroquinolone antibacterial agent, was evaluated in multiple strains of mice and rats. In 5 separate studies, mice and rats that were orally administered up to 1,000 mg/kg of clinafloxacin from 9 to 104 weeks developed dose-related cecal dilatation and deaths that were attributable to cecal torsion. Cecal rupture was observed in association with torsion. Although cecal dilatation is commonly observed in rodents given antibacterials such as fluoroquinolones, cecal torsion has not been a reported sequelae to dilatation. PMID- 11026599 TI - A mechanistic study of ovarian carcinogenesis induced by nitrofurazone using rasH2 mice. AB - In order to clarify whether the ovarian tumors induced in a long-term carcinogenicity study of nitrofurazone (NF) in mice can be also produced in a short-term model using transgenic (Tg) mice carrying the human c-Ha-ras gene (rasH2 mice), the following 3 experiments were performed. In experiment 1, both rasH2 mice and their wild CB6F1 littermates carrying no c-Ha-ras gene (non-Tg mice) that were fed a diet containing 500 to 1,000 ppm NF for 7 weeks demonstrated ovarian atrophy characterized by decreased labeling indices (LIs) for proliferating cell nuclear antigen (PCNA) in granulosa cells. In experiment 2, increased numbers of atretic follicles and decreased PCNA LIs in granulosa cells were recognized in rasH2 mice given diets containing 250 or 500 ppm NF for 26 weeks, but no tumor induction was grossly observed. In experiment 3, similar ovarian atrophy was observed in association with increased serum luteinizing hormone (LH) levels in both rasH2 and non-Tg mice given diet containing 1,000 ppm NF for 11 days. These results indicate that long-term NF treatment induces ovarian tumors in mice, possibly by continuous stimulation with gonadotropins such as LH via a negative-feedback phenomenon secondary to ovarian atrophy (as the tumor-induction mechanism), although we could not completely rule out a genotoxic mechanism. PMID- 11026600 TI - Short-term pulmonary response to inhaled JP-8 jet fuel aerosol in mice. AB - B6.A.D. (Ahr(d)/Nat(s)) mice were utilized to investigate the short-term pulmonary response to JP-8 jet fuel (JP-8) aerosol inhalation. Mice were nose only exposed to atmospheres of 0 to 118 mg/m3 for 1 h/d over a period of 7 days to further test the hypothesis that JP-8 concentrations below the permissible exposure level (PEL) of 350 mg/m3 will induce lung injury. At 24 to 30 hours after the final exposure, pulmonary function and respiratory permeability were measured on anesthetized mice and then randomly assigned for bronchoalveolar lavage or histopathology. Bronchoalveolar lavage fluid (BALF) was analyzed for total protein, lactic dehydrogenase (LDH), N-acetyl-beta-D-glucosaminidase (NAG), and cytology. Respiratory permeability increases were observed following doses of 48 and 118 mg/m3 and were supported by concomitant BALF increases in total protein and LDH. Conversely, NAG and alveolar macrophage levels decreased following the same exposure concentrations. Morphological lung injury was characterized by the targeting of bronchiolar epithelium and consisted of perivascular edema, Clara cell vacuolization, and necrosis. Alveolar injury included sporadic pulmonary edema, intra-alveolar hemorrhage, and alterations in type II epithelial cells. These results indicate that repeated inhalation of aerosolized JP-8 induces physiological, biochemical, cellular, and morphological lung injury. This study also provides evidence for the reevaluation of the 350 mg/m3 PEL for more volatile petroleum distillates with regard to respirable aerosols. PMID- 11026601 TI - Immunohistochemical localization of transforming growth factor alpha in chemically induced rat hepatocellular carcinomas with reference to differentiation and proliferation. AB - Hepatocellular carcinomas (HCCs) were induced in male Fischer 344 rats with dietary 3'-methyl-4-(dimethylamino)-azobenzene treatment and were classified into solid, glandular (well- or poorly differentiated), and trabecular types. Investigation of cell proliferation kinetics and immunohistochemical localization of transforming growth factor alpha (TGF-alpha) demonstrated all solid (n = 24) and poorly differentiated glandular type (n = 6) HCCs to have TGF-alpha-positive nuclei. Nuclear staining of TGF-alpha was also observed in 13 of 28 (46%) trabecular-type HCCs, whereas 12 (43%) exhibited cytoplasmic staining, and 3 (11%) were negative. As for well-differentiated glandular HCCs, 7 of 20 (35%) were positively stained in their nucleus, another 7 (35%) demonstrated antibody binding in the cytoplasm, and 6 (30%) were negative. The order for growth rate evaluated by bromodeoxyuridine (BrdU) labeling was solid (38.22%), poorly differentiated glandular (26.82%), trabecular (7.98%), and well-differentiated glandular (2.57%) types. For trabecular HCCs with nuclear, cytoplasmic, or negative TGF reactions, values were 13.39% (n = 13), 3.61% (n = 12), and 2.01% (n = 3), respectively. Likewise, BrdU-labeling indices for the counterpart groups of well-differentiated glandular type HCCs were 4.53, 1.91, and 1.29%, respectively. The results indicate that TGF-alpha expression might be linked to histopathological differentiation and cell proliferation in rat HCCs. PMID- 11026603 TI - Microscopic changes with acetic acid and sodium hydroxide in the rabbit low volume eye test. AB - Differences in ocular irritancy have been hypothesized to reflect differences in the extent of initial injury. Although differences in the processes leading to tissue damage may exist, extent of injury is believed to be the principal factor determining final outcome of ocular irritation. Previous studies characterizing the pathology of surfactant-induced ocular irritation support this premise. The purpose of this study was to begin to determine the applicability of this premise in terms of nonsurfactants; we planned to accomplish this by assessing the ocular irritancy of different concentrations of an acid and an alkali. Ten microliters of 3 or 10% acetic acid (C2H4O2) or 2 or 8% sodium hydroxide (NaOH) were directly applied to the cornea of the right eye of each test rabbit. Untreated left eyes served as the controls. Eyes and eyelids were macroscopically examined for signs of irritation beginning 3 hours after dosing and periodically until recovery or day 35. Eyes and eyelids from animals in each group were collected for microscopic examination after 3 hours and on days 1, 3, and 35. The macroscopic and microscopic changes were consistent with slight (3% C2H4O2), mild (2% NaOH, 10% C2H4O2), and severe (8% NaOH) irritancy. The spectra of changes were similar to those previously reported for surfactants of differing types and irritancies. As with surfactants, as the extent of initial injury increased, the intensity and duration of the subsequent responses increased. These results indicate that our hypothesis also applies to nonsurfactants. The results also support our belief that the initial extent of injury associated with ocular irritation may be used to predict the subsequent responses and final outcome. Finally, our results further indicate that such an approach may be applicable to the development of alternative assays that are based on either injury to ex vivo eyes or injury to an in vitro corneal equivalent system. PMID- 11026602 TI - Quantitative characterization of acid- and alkali-induced corneal injury in the low-volume eye test. AB - Defining the extent of initial injury has proven to be a useful basis for differentiating the ocular irritation potential of surfactants; however, the applicability of this method to other types of irritants has not been demonstrated. In the following studies we characterized the extent of corneal injury following exposure to different concentrations of acetic acid and sodium hydroxide (NaOH) in the rabbit low-volume eye test. Groups of rabbits received 3% acetic acid, 10% acetic acid, 2% NaOH, or 8% NaOH and were evaluated in vivo by macroscopic and in vivo confocal microscopic examination and postmortem using a live/dead staining kit and scanning laser confocal microscopic examination. Quantitative assessment of macroscopic scores, corneal surface epithelial cell size, corneal epithelial thickness, corneal thickness, depth of stromal injury, corneal light scattering (confocal microscopy through focusing, CMTF), and number of dead cells was conducted at various times, including the following: at 3 hours and at 1, 3, 7, 14, and 35 days. Based on macroscopic scores, the order of ocular irritancy potential was 3% acetic acid < 2% NaOH < 10% acetic acid < 8% NaOH. Evaluation of the quantitative in vivo and postmortem microscopic live/dead data revealed a slight decrease in epithelial thickness and an increase in dead epithelial cell numbers with 3% acetic acid. With 2% NaOH, significant focal changes in epithelial cell size, epithelial thickness, corneal thickness, and number of dead surface epithelial cells occurred at 3 hours and at 1 day, with injury to only a very small number of corneal stromal keratocytes, despite the presence of epithelial denudation. Changes with 10% acetic acid were similar to those noted with 2% NaOH at 3 hours and 1 day, but these changes were more diffuse and included stromal injury to a depth of 7.2 +/- 9.3% of the corneal thickness, with significant numbers of dead keratocytes. Eight percent NaOH, on the other hand, caused focally extensive injury that averaged 26.3 +/- 18.4% of the corneal thickness at 1 day, with significant light scattering from the cornea, which did not return to normal by 35 days postinjury. Overall, these data indicate that ocular irritation as a result of acetic acid and NaOH was associated with changes similar to those observed with surfactants (ie, slight irritants damage the corneal epithelium, mild and moderate irritants damage the corneal epithelium and anterior stromal cells, and severe irritants damage the corneal epithelium and deep stroma). To our knowledge, this is the first time that the ocular irritation potential for different types of materials (acid/alkali, surfactants) has been shown to be primarily dependent on the initial area and depth of injury. PMID- 11026604 TI - Evaluation of plasma von Willebrand factor as a biomarker for acute arterial damage in rats. AB - Plasma von Willebrand factor (vWF) was evaluated as a potential biomarker of acute arterial damage in rats after a vasotoxic dose of the dopaminergic vasodilator, fenoldopam (FP). Male Sprague-Dawley rats were given FP or isotonic saline by subcutaneous injection, and plasma vWF was measured at 2, 6, and 24 hours after challenge. Mean plasma vWF values increased in FP-treated rats compared to controls at 2 hours (167 vs 122%; p < 0.05) and 6 hours postdose (172 vs 130%; p < 0.01) but were comparable to control values after 24 hours. Mesenteric arterial lesions were observed microscopically in all FP-treated rats 24 hours postdose but were not present in rats at 1, 2, 4, 6, or 8 hours after FP challenge. Further, plasma vWF concentrations increased in saline-treated rats after only the minimal perturbation of repeated venipuncture. These results indicate an early, minimal, and transient release of vWF that precedes the onset of morphologically evident vascular damage. The minimal increases in plasma vWF concentrations were of limited predictive value, may be more reflective of an acute-phase reactant response, and were not considered a reliable biomarker of acute FP-induced arterial damage in the rat. PMID- 11026605 TI - Dandy-Walker like malformation in a Fischer-344 rat. AB - A spontaneous cerebellar malformation was found in a 32-day-old male Fischer 344 rat. The cerebellar malformation was composed of a vermis defect and markedly dilated fourth ventricle. The cerebellar hemispheres were separated, with the left hemisphere being smaller than the right one. Degenerative/inflammatory lesions consisting of macrophage/lymphocyte infiltration, spheroidal calcium depositions, and astrocytic gliosis were seen adjacent to the cerebral aqueduct. Abnormally arranged hyperplastic ependymal cells were observed beneath the fourth ventricle in the medulla oblongata. The gross findings of the present case resemble those of the human Dandy-Walker malformation. While a precise mechanism remains to be elucidated, degenerative/inflammatory lesions in the present rat may be involved in the pathogenesis of this malformation. PMID- 11026606 TI - Ribose cysteine protects against acetaminophen-induced hepatic and renal toxicity. AB - Ribose cysteine (RibCys) is a cysteine prodrug that increases both hepatic and renal glutathione with documented antagonism of acetaminophen (APAP)-induced hepatotoxicity. To determine if RibCys could also protect against APAP-induced kidney damage, mice were injected with APAP (600 mg/kg) or APAP and RibCys (1.0 g/kg) (APAP/RIB) followed by additional RibCys injections 1 and 2 hours later. Mice were euthanatized 10-12 hours after APAP administration, and liver and kidney toxicity were assessed by plasma sorbitol dehydrogenase (SDH) activity and blood urea nitrogen (BUN), respectively, and by histopathology. APAP treatment resulted in elevation of SDH activity and BUN to 2,490 U/ml and 47 mg/dl, respectively. By contrast, SDH and BUN values for APAP/RIB-treated mice were not different from controls, 0 U/ml and 31 mg/dl, respectively. Histopathologic examination revealed moderate to severe hepatic centrilobular necrosis in 9/11 and renal proximal tubular necrosis in 10/11 APAP-treated mice. However, no evidence of hepatic or renal toxicity was noted in any of the 12 APAP/RIB-treated mice. Utilizing the same treatment regimen, APAP covalent binding to hepatic and renal cytosolic proteins was assessed 4 hours after APAP challenge. RibCys cotreatment decreased covalent binding to the 58-kDa acetaminophen-binding protein in both liver and kidney. RibCys decreased both toxicity and covalent binding after APAP administration, and in addition to protecting the liver, this cysteine prodrug can also effectively protect the kidney from APAP-induced injury. PMID- 11026607 TI - Neoplasia in zebrafish (Danio rerio) treated with 7,12-dimethylbenz[a]anthracene by two exposure routes at different developmental stages. AB - Using zebrafish, Danio rerio, initial pioneering work in the 1960s revealed carcinogen responsiveness of fish, yet very few subsequent tumorigenesis investigations have utilized this species. We exposed embryos (60 hours postfertilization) and fry (3 week posthatch) to 7,12-dimethylbenz[a]anthracene (DMBA) by immersion in aqueous solutions for 24 hours, at concentrations of 0-1 or 0-5 ppm (mg/L), respectively. Juvenile zebrafish 2 months posthatch were fed a diet containing 0-1,000 ppm DMBA for 4 months. Fish were sampled for histologic evaluation at 7-12 months after the onset of carcinogen treatment. Fry were most responsive to DMBA and showed the widest diversity of target tissues and histologic types of neoplasia, having several types of epithelial, mesenchymal, and neural neoplasia. The principal target tissues for carcinogenic response were liver following embryo or fry exposure, with gill and blood vessel the second and third most responsive tissues in fry. Intestine was the primary target and gill a secondary target in fish that received dietary DMBA as juveniles. These studies indicate that young zebrafish are most responsive to DMBA, showing a greater diversity of neoplasm types than rainbow trout. Thus, zebrafish are a valuable model system in which to study mechanistic aspects of the carcinogenesis process. PMID- 11026608 TI - Neoplasia in zebrafish (Danio rerio) treated with N-methyl-N'-nitro-N nitrosoguanidine by three exposure routes at different developmental stages. AB - We exposed embryos (83 hours postfertilizaton) and fry (3 weeks posthatch) to N methyl-N'-nitro-N-nitrosoguanidine (MNNG) by immersion in aqueous solutions of 0 10 ppm for 1 hour (embryo) or 0-2 ppm for 24 hours (fry). Zebrafish embryos were microinjected with MNNG at levels of 0 or 96 ng/egg. Diets containing 0-2,000 ppm MNNG were fed to juvenile zebrafish for 3 months beginning at 2 months posthatch. Fish were sampled for histopathologic study at 6-12 months after initiation of carcinogen exposure. Embryos and fry were both quite responsive to MNNG; however, juvenile zebrafish were remarkably refractory to MNNG-induced neoplasia. Principal target organs in zebrafish treated as embryos with MNNG were liver and testis, with hepatocellular adenoma the most prevalent hepatic neoplasm. A variety of mesenchymal neoplasms occurred in zebrafish following embryo exposure to MNNG, including chondroma, hemangioma, hemangiosarcoma, leiomyosarcoma, and rhabdomyosarcoma. Testis and blood vessels were primary target organs for MNNG following fry exposure, with seminoma, hemangioma, hemangiosarcoma, and various other epithelial and mesenchymal neoplasms occurring. The zebrafish is a responsive, cost-effective lower vertebrate model system in which to study mechanisms of carcinogenesis. PMID- 11026609 TI - Fulminant hepatocyte apoptosis in vivo following microcystin-LR administration to rats. AB - Microcystin-LR (MCLR) is a cyanobacterial toxin responsible for human and livestock deaths worldwide. MCLR has also been implicated as a contributing factor in hepatocellular carcinoma. Following absorption, MCLR is taken up via a hepatocyte-specific bile acid carrier. Inside hepatocytes, MCLR selectively binds to protein phosphatases 1 and 2A, resulting in rapid, massive liver damage. However, the apoptotic nature of this toxicosis in rats has not been fully characterized as such at appropriate time points utilizing light and electron microscopy, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and electrophoresis of hepatic DNA. Rats were administered intraperitoneal saline or MCLR at 500 microg/kg (0.5 micromol/kg) and necropsied at 3 or 9 hours. Light microscopy at 3 hours revealed massive, widespread apoptotic necrosis of the majority of hepatocytes. Hepatocytes were rounded and disassociated, with cell shrinkage, increased eosinophilia, and margination of nuclear chromatin or pyknosis. The apoptotic index increased from 0.03% +/- 0.02% in controls to 205% +/- 12% in MCLR-treated animals (p < or = 0.0001). At 3 hours, transmission electron microscopy revealed hepatocellular changes typical of apoptotic necrosis: rounding and disassociation of hepatocytes, loss of microvilli, and margination and condensation of nuclear chromatin. Laddering of hepatic DNA by electrophoresis and widespread TUNEL staining of hepatocytes were consistent with apoptosis. These results demonstrate that in rats, hepatic damage caused by MCLR is due to extremely rapid induction and progression of apoptosis in virtually every hepatocyte in the liver. This model of fulminant hepatic necrosis should be useful for increased characterization and understanding of the relationship between protein phosphatase inhibition and apoptosis. PMID- 11026610 TI - Inhalation studies in laboratory animals--current concepts and alternatives. AB - Highly standardized and controlled inhalation studies are required for hazard identification to make test results reproducible and comparable and to fulfill general regulatory requirements for the registration of new drugs, pesticides, or chemicals. Despite significant efforts, the results of inhalation studies have to be analyzed judiciously due to the great number of variables. These variables may be related to technical issues or to the specific features of the animal model. Although inhalation exposure of animals mimics human exposure best, ie, error prone route-to-route extrapolations are not necessary, not all results obtained under such very rigorous test conditions may necessarily also occur under real life exposure conditions. Attempts are often made to duplicate as closely as possible these real-life exposure conditions of humans in appropriate bioassays. However, this in turn might affect established baseline data, rendering the interpretation of new findings difficult. In addition, specific use patterns, eg, of inhalation pharmaceuticals or pesticide-containing consumer products, may impose test agent-specific constraints that challenge traditional approaches. Moreover, specific modes of action of the substance under investigation, the evaluation of specific endpoints, or the clarification of equivocal findings in common rodent species may require exposure paradigms or the use of animal species not commonly used in inhalation toxicology. However, particularly in inhalation toxicology, the choice of animal models for inhalation toxicity testing is usually based on guideline requirements and practical considerations, such as exposure technology, expediency, and previous experience rather than validity for use in human beings. Larger animal species, apart from the welfare aspects, may require larger inhalation chambers to accommodate the animals, but for technical reasons and the difficulty of generating homogeneous exposure atmospheres in such inhalation chambers, this may jeopardize the outcome of the study. Some of the many variables and possible artifacts likely to occur in animal inhalation studies are addressed in this paper. PMID- 11026611 TI - Chronic inhalation studies with mainstream cigarette smoke in rats and mice. PMID- 11026612 TI - A case report of invasive pituitary tumors containing alpha-melanocyte stimulating hormone arising from the pars distalis in F344 rats. PMID- 11026613 TI - Current methods for the identification and quantitation of ceramides: an overview. AB - Ceramides are key compounds in the metabolism of sphingolipids and are emerging as important second messengers for various cellular processes including cell cycle arrest, differentiation, senescence, apoptosis, and others. Because of their important biological functions, exact analysis of their molecular species and concentrations is crucial for elucidating their function and metabolism. Toward this goal, several methods have been developed for the identification and quantitation of cellular ceramide levels. Methods have been developed utilizing thin-layer or high-performance liquid chromatography. Mass spectrometry also has become increasingly utilized. The Escherichia coli diacylglycerol kinase assay is one of the most frequently used techniques for ceramide quantitation. This review presents a current summary of methods used for the identification and quantitation of ceramides. PMID- 11026614 TI - Autoxidation of methyl linoleate: identification of the bis-allylic 11 hydroperoxide. AB - Based on the understanding of lipid peroxidation as a free radical chain reaction, over 50 yr ago the three primary products of linoleic acid autoxidation were predicted to be the 9-, 11-, and 13-hydroperoxides. The 9- and 13 hydroperoxides were found at the time, but formation of 11-hydroperoxylinoleate or any other bis-allylic fatty acid hydroperoxide has not been reported heretofore as a product of lipid peroxidation reactions. In vitamin E-controlled autoxidation of methyl linoleate, the 11-hydroperoxy derivative was identified as the next most prominent primary peroxidation product after the 9- and 13 hydroperoxides. It was present in approximately 5-10% of the abundance of the 9- or 13-hydroperoxide. The structures of 1l-hydroperoxylinoleate and its 11-hydroxy derivative were established by high-pressure liquid chromatography, ultraviolet spectroscopy, gas chromatography mass spectroscopy, and 1H nuclear magnetic resonance spectroscopy. The 11-hydroperoxide was not detectable in the absence of (alpha-tocopherol, indicating that efficient trapping of the 11-peroxyl radical as the hydroperoxide is critical to permitting its accumulation. PMID- 11026615 TI - Synthesis of 9,12-dioxo-10(Z)-dodecenoic acid, a new fatty acid metabolite derived from 9-hydroperoxy-10,12-octadecadienoic acid in lentil seed (Lens culinaris Medik.) AB - The previously unknown linoleic acid peroxidation product 9,12-dioxo-10(Z) decenoic acid (Z5) was detected in lentil seed flour (Lens culinaris Medik.) by electron impact mass spectrometry (EI-MS) after derivatization with pentafluorobenzyl-hydroxylamine-hydrochloride, methylation of acidic groups with diazomethane, and protection of hydroxylic groups with N-methyl-N-trimethylsilyl trifluoroacetamide. The structure of the natural product was confirmed by synthesis of Z5, 9,12-dioxo-l0(E)-decenoic acid, and derivatives. EI-MS, nuclear magnetic resonance and gas chromatographic data of these compounds and synthetic intermediates are discussed. PMID- 11026616 TI - Formation of 8-oxo-2'-deoxyguanosine in the DNA of human diploid fibroblasts by treatment with linoleic acid hydroperoxide and ferric ion. AB - Lipid peroxides are suggested to be related to the occurrence of a variety of diseases including cancer and atherosclerosis. We examined whether lipid peroxides cause oxidative damage to DNA in intact cells. Linoleic acid hydroperoxide (LOOH) and ferric chloride were used at concentrations at which separate treatment had no effect on the formation of 8-oxo-2'-deoxyguanosine (8 oxodG) in DNA or the survival rate of cultured human diploid fibroblasts, TIG-7. The amount of 8-oxodG in the cellular DNA increased significantly when TIG-7 cells were treated concurrently with LOOH and ferric chloride. In a LOOH concentration-dependent manner 8-oxodG was formed. However, no significant induction of the activities of superoxide dismutases, catalase, or glutathione peroxidase was observed under these conditions. The formation of 8-oxodG by lipid hydroperoxides seems to be due to the generation of reactive species other than superoxide radicals and hydrogen peroxide. These results indicate that some species formed during the reaction of lipid hydroperoxides with ferric ion can cause oxidative damage to DNA. PMID- 11026618 TI - Comparison of growth and fatty acid metabolism in rats fed diets containing equal levels of gamma-linolenic acid from high gamma-linolenic acid canola oil or borage oil. AB - We have utilized transgenic technology to develop a new source of gamma-linolenic acid (GLA) using the canola plant as a host. The aim of the present study was to compare the growth and fatty acid metabolism in rats fed equal amounts of GLA obtained from the transgenic canola plant relative to GLA from the borage plant. Young male Sprague-Dawley rats (n = 10/group) were randomized and fed a purified AIN93G diet (10% lipid by weight) containing either a mixture of high GLA canola oil (HGCO) and corn oil or a control diet containing borage oil (BO) for 6 wk. GLA accounted for 23%, of the triglyceride fatty acids in both diets. Growth and diet consumption were monitored every 2-3 d throughout the study. At study termination, the fatty acid composition of the liver and plasma phospholipids was analyzed by gas chromatography. The growth and diet consumption of the HGCO group were similar to the BO group. There were no adverse effects of either diet on the general health or appearance of the rats, or on the morphology of the major organs. There was no significant difference between the diet groups for total percentage of n-6 polyunsaturated fatty acids present in either the total or individual phospholipid fractions of liver or plasma. The relative percentage of GLA and its main metabolite, arachidonic acid, in each phospholipid fraction of liver or plasma were also similar between groups. The percentage of 18:2n-6 in liver phosphatidylethanolamine and phosphatidylinositol/serine was higher (P < 0.05) and 22:5n-6 was lower in the HGCO group than the BO group. This finding could be attributed to the higher 18:3n-3 content in the HGCO diet than the BO diet. Results from this long-term feeding study of rats show for the first time that a diet containing transgenically modified canola oil was well-tolerated, and had similar biological effects, i.e., growth characteristics and hepatic metabolism of n-6 fatty acids, as a diet containing borage oil. PMID- 11026617 TI - Lipid peroxidation as determined by plasma isoprostanes is related to disease severity in mild asthma. AB - Oxidlative stress is believed to play an important role in the pathophysiology of asthma. Recently discovered F2-isoprostanes, of which 8-iso-PGF2alpha is the most well-known isomer, have emerged as the most reliable marker of in vivo oxidative stress. The aim of this study was to examine 8-iso-PGF2alpha as a biomarker of oxidative stress in mild asthma in relation to endogenous and dietary antioxidant protection. Total (free and esterified) plasma 8-iso-PGF2alpha, plasma dietary antioxidants (vitamins E and C, Beta-carotene, Zn, and Se), and erythrocyte antioxidant enzyme activities (glutathione peroxidase and superoxide dismutase) were measured in 15 mild asthmatics and 15 age-and sex-matched controls. Total plasma 8-iso-PGF2alpha levels [median (quartile 1 - quartile 3)] were significantly increased in the asthmatics [213 pg/mL (122-455) vs. 139 pg/mL (109 174), P= 0.042]. The 8-iso PGF2alpha levels were found to be associated with clinical asthma severity (P = 0.044) and inhaled corticosteroid use (P = 0.027) in asthmatics. No differences were observed in the plasma dietary antioxidant vitamins. The asthmatics had significantly lower plasma levels of Zn (P = 0.027) and Se (P = 0.006). Plasma Se correlated negatively with 8-iso-PGF2alpha (r = 0.725, P= 0.002). No differences between the groups were observed for glutathione peroxidase or superoxide dismutase, however, superoxide dismutase activity was negatively associated with asthma severity (P = 0.042). In conclusion, oxidative stress is increased in mildly asthmatics, as reflected by increased plasma levels of 8-iso-PGF2alpha and a deficiency in plasma Zn and Se. The isoprostane 8-iso PGF2alpha may provide a useful tool in intervention studies aimed at improving clinical status in asthma. PMID- 11026619 TI - Calcium deficiency modifies polyunsaturated fatty acid metabolism in growing rats. AB - Fatty acid desaturase activities were determined in liver microsomes from calcium deficient rats and compared to calcium-sufficient ones. The calcium-deprived diet (0.5 g/kg) administered for 60 d caused a 30% inhibition in the delta5 desaturase activity and a 45-55% decrease in delta6 and delta9, respectively, facts that cannot be attributed to a reduction in food intake. In vitro addition of calcium, ethyleneglycol-bis(Beta-aminoethyl ether)N,N-tetraacetic acid, and/or cytosol fractions from control or calcium-deficient rats to microsomes from both groups of animals indicates that the reduced desaturase capacities would be the consequence of an indirect effect of calcium deprivation. The present work shows that the reduced unsaturated fatty acid biosynthesis might be the result of modifications in the physicochemical properties of microsomal membranes. Such changes could also be derived from the inhibition of phospholipase A2 activity induced by calcium deficiency. PMID- 11026620 TI - Response of plasma lipids to dietary cholesterol and wine polyphenols in rats fed polyunsaturated fat diets. AB - This experiment was designed to evaluate the effects of dietary red wine phenolic compounds (WP) and cholesterol on lipid oxidation and transport in rats. For 5 wk, weanling rats were fed polyunsaturated fat diets (n-6/n-3 = 6.4) supplemented or not supplemented with either 3 g/kg diet of cholesterol, 5 g/kg diet of WP, or both. The concentrations of triacylglycerols (TAG, P < 0.01) and cholesterol (P < 0.0002) were reduced in fasting plasma of rats fed cholesterol despite the cholesterol enrichment of very low density lipoprotein + low density lipoprotein (VLDL + LDL). The response was due to the much lower plasma concentration of high density lipoprotein (HDL) (-35%, P < 0.0001). In contrast, TAG and cholesteryl ester (CE) accumulated in liver (+120 and +450%, respectively, P < 0.0001). However, the cholesterol content of liver microsomes was not affected. Dietary cholesterol altered the distribution of fatty acids mainly by reducing the ratio of arachidonic acid to linoleic acid (P < 0.0001) in plasma VLDL + LDL (-35%) and HDL (-42%) and in liver TAG (-42%), CE (-78%), and phospholipids (-28%). Dietary WP had little or no effect on these variables. On the other hand, dietary cholesterol lowered the alpha-tocopherol concentration in VLDL + LDL ( -40%, P < 0.003) and in microsomes (-60%, P < 0.0001). In contrast, dietary WP increased the concentration in microsomes (+21%, P < 0.0001), but had no effect on the concentration in VLDL + LDL. Cholesterol feeding decreased (P < 0.006) whereas WP feeding increased (P < 0.0001) the resistance of VLDL + LDL to copper-induced oxidation. The production of conjugated dienes after 25 h of oxidation ranged between 650 (WP without cholesterol) and 2,560 (cholesterol without WP) micromol/g VLDL + LDL protein. These findings show that dietary WP were absorbed at sufficient levels to contribute to the protection of polyunsaturated fatty acids in plasma and membranes. They could also reduce the consumption of alpha tocopherol and endogenous antioxidants. The responses suggest that, in humans, these substances may be beneficial by reducing the deleterious effects of a dietary overload of cholesterol. PMID- 11026621 TI - The effect of low alpha-linolenic acid diet on glycerophospholipid molecular species in guinea pig brain. AB - The changes in guinea pig brain (cerebrum) glycerophospholipid molecular species resulting from a low-alpha linolenic acid (ALA) diet are described. Two groups of six guinea pigs were raised from birth to 16 wk of age on either an n-3 deficient diet containing 0.01 g ALA/100 g diet or n-3 sufficient diet containing 0.71 g ALA/100 g diet. Molecular species of diradyl glycerophosphoethanolamine (GroPEtn), glycerophosphocholine, glycerophosphoserine, and glycerophosphoinositol were analyzed by high-performance liquid chromatography with on-line electrospray ionization mass spectrometry (HPLC/ESI/MS). Alkenylacyl GroPEtn species were determined by comparing spectra before and after mild acid treatment while diacyl- and alkylacyl species were distinguished by HPLC/ESI/MS. The proportions of phospholipid classes and of the diradyl GroPEtn subclasses were not altered by diet changes. The main polyunsaturated molecular species of diradyl GroPEtn subclasses and of phosphatidyl choline and phosphatidylserine (PtdSer) contained 16:0, 18:0, or 18:1 in combination with docosahexaenoic acid (DHA, 22:6n-3), docosapentaenoic (DPA, 22:5n-6), or arachidonic acid (ARA, 20:4n 6). A significant proportion of DPA containing species were present in both diet groups, but in n-3 fatty acid deficiency, the proportion of DPA increased and DHA was primarily replaced by DPA. The combined value of main DHA and DPA containing species in the n3 deficient group ranged from 91-111% when compared with the n-3 sufficient group, indicating a nearly quantitative replacement. The n-3 fatty acid deficiency did not lower the content of ARA containing molecular species of PtdSer of the guinea pig brain as reported previously for the rat brain. The molecular species of phosphatidylinositol were not altered by n-3 fatty acid deficiency. The present data show that the main consequence of a low ALA diet is the preferential replacement of DHA-containing molecular species by DPA containing molecular species in alkenylacyl- and diacyl GroPEtn and PtdSer of guinea pig brain. PMID- 11026622 TI - Dietary docosahexaenoic acid affects stearic acid desaturation in spontaneously hypertensive rats. AB - Docosahexaenoic acid (DHA, 22:6n-3) is an n-3 polyunsaturated fatty acid which attenuates the development of hypertension in spontaneously hypertensive rats (SHR). The effects of DHA on delta-9-desaturase activity in hepatic microsomes and fatty acid composition were examined in young SHR. Two groups of SHR were fed either a DHA-enriched diet or a control diet for 6 wk. Desaturase activity and fatty acid composition were determined in hepatic microsomes following the dietary treatments. Delta-9-desaturase activity was decreased by 53% in DHA-fed SHR and was accompanied by an increase in 16:0 and a reduction in 16:1n-7 content in hepatic microsomes. The DHA diet also increased the levels of eicosapentaenoic acid (20:5n-3) and DHA. The n-6 fatty acid content was also affected in DHA-fed SHR as reflected by a decrease in gamma-linolenic acid (18:3n-6), arachidonic acid (20:4n-6), adrenic acid (22:4n-6), and docosapentaenoic acid (22:5n-6). A higher proportion of dihomo-gamma-linolenic acid (20:3n-6) and a lower proportion of 20:4n-6 is indicative of impaired delta-5-desaturase activity. The alterations in fatty acid composition and metabolism may contribute to the antihypertensive effect of DHA previously reported. PMID- 11026623 TI - Secretion of hepatic lipase by perfused liver and isolated hepatocytes. AB - Hepatic lipase is found in liver and in adrenal glands and ovaries. Because in adult rats, neither adrenals nor ovaries synthesize this enzyme, it is assumed that the liver is the origin of their hepatic lipase. Our aim was to study the secretion of hepatic lipase by the liver. We observed that plasma of both fed and fasted rats contained hepatic lipase activity. This activity was significantly correlated with that in the liver. Isolated livers, perfused with heparin-free medium, secreted fully active hepatic lipase to the perfusate. The addition of heparin resulted in a rapid and larger release of hepatic lipase to the perfusate. In isolated hepatocytes, heparin did not affect the secretion of hepatic lipase mass, although it increased the stability of the enzyme activity. To study the degradation of hepatic lipase by hepatocytes, protein synthesis was blocked with cycloheximide, and both secreted and intracellular hepatic lipases were analyzed by Western blotting. We observed that the amount of hepatic lipase secreted equaled the decrease of intracellular mass. The total mass of the enzyme (inside and outside the cells) remained constant, at least for 90 min. In the next experiment, 0.7 nM 125I-hepatic lipase was added to hepatocyte suspensions, and the appearance of trichloracetic acid-soluble products was analyzed. Only 12% of the radioactivity added was associated with the cells after 90 min of incubation, and less than 2% of the hepatic lipase added was degraded. Although the association was decreased in the presence of heparin, the amount of 125I hepatic lipase degraded was not affected. Taking all these results into account, we propose a model for the continuous secretion of hepatic lipase by the liver. PMID- 11026624 TI - Study of individual trans- and cis-16:1 isomers in cow, goat, and ewe cheese fats by gas-liquid chromatography with emphasis on the trans-delta3 isomer. AB - Low-temperature gas-liquid chromatography (GLC) was applied to study the distribution profiles of isomeric trans- and cis-hexadecenoic acids in ruminant (cow, goat, and ewe) milk fat after their fractionation by argentation thin-layer chromatography (Ag-TLC). The fat was extracted from cheeses (12 samples of each species), the most common foods made with goat and ewe milks. The predominant trans-16:1 isomer is palmitelaidic acid (the delta9 isomer), but it does not exceed one-third of the total group, which itself represents 0.17% (cow), 0.16% (goat), and 0.26% (ewe) of the total fatty acids. The trans-delta3 16:1 isomer, which is reported for the first time in ruminant lipids and which likely comes from the animals' feed, is present at a level of ca. 10% of the trans-16:1 acid group. Otherwise, all isomers with their ethylenic bond between positions delta4 and delta14 are observed in the three species studied, roughly showing the same relative distribution pattern. Quantitatively, the trans-16:1 isomers only represent ca. 5% of the sum of the trans-16:1 plus trans-18:1 isomers, and they appear of little importance in comparison. It is inferred from this and recent studies that some previously reported data that were established for consumption assessments dealt in fact mainly with iso-17:0 acid, which was confused with (and added to) trans-delta9 (palmitelaidic) acid; consequently, these results were large over-estimates. Regarding the cis-16:1 acids, the delta9 isomer is the prominent constituent as expected, but the second-most important isomer is the delta13 isomer. It does not appear that trans-16:1 isomers are from ruminant milk fats of great nutritional importance as compared with trans-18:1 isomeric acids. As for trans18:1 isomers, the combination Ag-TLC/GLC is a necessary procedure to quantitate trans-16:1 acids accurately and reliably. Ag-TLC allows removal of interfering branched 17:0 acids and cis-16:1 acids, and low-temperature GLC permits an accurate measurement of all individual isomers most of which with baseline resolution. PMID- 11026625 TI - Determination of the phospholipid/lipophilic compounds ratio in liposomes by thin layer chromatography scanning densitometry. AB - The determination of the ratio of phospholipid/lipophilic compounds in liposomes was achieved after thin-layer chromatography (TLC) by measuring the spot intensities of dipalmitoyl phosphatidylcholine and the lipophilic compound. The liposome components under study were separated on one TLC plate, developed in two steps, and detected after charring the plate with specific visualization reagents. The method shows good reproducibility and provides a simple way to quantify the level of lipophilic compound incorporated in the liposome bilayer. PMID- 11026626 TI - Deuterium uptake and plasma cholesterol precursor levels correspond as methods for measurement of endogenous cholesterol synthesis in hypercholesterolemic women. AB - To assess the validity of two techniques used to measure human cholesterol synthesis, the rate of uptake of deuterium (D) into plasma free cholesterol (FC), and plasma cholesterol precursor (squalene, lanosterol, desmosterol and lathosterol) levels were compared in 14 women [65-71 yr with low density lipoprotein-cholesterol (LDL-C) > or = 3.36 mmol x L(-1)]. Subjects consumed each of six diets for 5-wk periods according to a randomized crossover design. The experimental diets included a baseline diet (39% energy as fat, 164 mg chol x 4.2 MJ(-1)) and five reduced-fat diets (30% of energy as fat), where two-thirds of the fat was either soybean oil; squeeze, tub or stick margarines; or butter. Fractional and absolute synthesis rates (FSR and ASR) of FC were determined using the deuterium incorporation (DI) method, while cholesterol precursor levels were measured using gas-liquid chromatography. Data were pooled across diets for each variable and correlation coefficients were calculated to determine if associations were present. There was good agreement among levels of the various cholesterol precursors. In addition, FSR in pools/d (p x d(-1)) and ASR in grams/d (g x d(-1)) were strongly associated with lathosterol (r= 0.72 and 0.71, P= 0.0001), desmosterol (r= 0.75 and 0.75, P = 0.0001), lanosterol (r = 0.67 and 0.67), and squalene (r = 0.69 and 0.68) when levels of the precursors were expressed as micromol x mmol(-1) C. Significant but lower correlations were observed between the D uptake and plasma cholesterol precursor levels when the latter were expressed in absolute amounts (micromol x L(-1)). The wide range of fatty acid profiles of the experimental diets did not influence the degree of association between methods. In conclusion, the DI method and levels of some cholesterol precursors correspond as methods for shortterm measurement of cholesterol synthesis. PMID- 11026627 TI - Antioxidant reactions of alpha-tocopherolhydroquinone. AB - Alpha-tocopherolhydroquinone (TQH2) is a product of alpha-tocopherol oxidation/reduction that exerts antioxidant effects in biological systems. TQH2 inhibited autoxidation of methyl linoleate initiated by peroxyl radicals derived from thermolysis of 2,2'-azobis(2,4-dimethylvaleronitrile) in acetonitrile. TQH2 oxidation yielded alpha-tocopherolquinone (TQ) as a major product and 2,3-epoxy alpha-tocopherolquinone and 5,6-epoxy-alpha-tocopherolquinone as minor products. Each TQH2 consumed approximately two peroxyl radicals in the course of the oxidation. The data suggest that TQH2 scavenges peroxyl radicals primarily by electron transfer to form TQ and secondarily by addition-elimination to form the epoxyquinones. PMID- 11026628 TI - Acellular vascular tissues: natural biomaterials for tissue repair and tissue engineering. AB - Various research groups around the world are actively investigating cardiovascular prostheses of biological origin. This review article discusses the need for such bioprosthetics and the potential role for natural tissues in cardiovascular applications such as cardiac valves and vascular grafts. Upon implantation, unmodified natural materials are subject to chemical and enzymatic degradation, seriously decreasing the life of the prosthesis. Therefore, methods such as glutaraldehyde and polyepoxide crosslinking treatments and dye-mediated photooxidation have been developed to stabilize the tissue while attempting to maintain its natural mechanical properties. Also, residual cellular components in a bioprosthetic material have been associated with undesired effects, such as calcification and immunological recognition, and thus have been the motivation for various decellularization processes. The effects of these stabilization and decellularization treatments on mechanical, biological and chemical properties of treated tissues have been investigated, specifically with regard to calcification, immunogenicity, and cytotoxicity concerns. Despite significant advances in the area of cardiovascular prostheses, there has yet to be developed a completely biocompatible, long-lasting implant. However, with the recent advent of tissue engineering, the possibility of applying selective cell seeding to naturally derived bioprosthetics moves us closer to a living tissue replacement. PMID- 11026629 TI - Biomaterial-microvasculature interactions. AB - The utility of implanted sensors, drug-delivery systems, immunoisolation devices, engineered cells, and engineered tissues can be limited by inadequate transport to and from the circulation. As the primary function of the microvasculature is to facilitate transport between the circulation and the surrounding tissue, interactions between biomaterials and the microvasculature have been explored to understand the mechanisms controlling transport to implanted objects and ultimately improve it. This review surveys work on biomaterial-microvasculature interactions with a focus on the use of biomaterials to regulate the structure and function of the microvasculature. Several applications in which biomaterial microvasculature interactions play a crucial role are briefly presented. These applications provide motivation and framework for a more in-depth discussion of general principles that appear to govern biomaterial-microvasculature interactions (i.e., the microarchitecture and physio-chemical properties of a biomaterial as well as the local biochemical environment). PMID- 11026630 TI - The effect of adsorbed fibrinogen, fibronectin, von Willebrand factor and vitronectin on the procoagulant state of adherent platelets. AB - Procoagulant (activated) platelets provide a site for assembly of the prothrombinase complex which can rapidly convert prothrombin into thrombin (a potent inducer of clot formation). Previously, we reported that adhesion of platelets to surfaces preadsorbed with blood plasma caused them to become procoagulant. In the present study we investigated the effect of adsorbed adhesion proteins (fibrinogen (Fg), fibronectin (Fn), von Willebrand factor (vWF) and vitronectin (Vn)) on the procoagulant activity of adherent platelets. Adsorbed Fn, vWF and Fg promoted platelet adhesion in the following order: Fn < vWF = Fg. However, these proteins promoted platelet activation (thrombin generation per adherent platelet) in the following order: Fg < Fn < vWF. Adsorption with a series of dilutions of normal plasma, serum, and plasmas deficient in or depleted of von Willebrand factor (de-vWF), fibronectin (de-Fn), vitronectin (de-Vn), or both vitronectin and fibronectin (de-VnFn) resulted in varied platelet adhesion, but little difference in platelet activation. However, preadsorption with dilute de-vWF plasma induced lower procoagulant activity than normal plasma. Preadsorption with normal plasma resulted in higher levels of platelet activation than preadsorption with Fg, suggesting that adsorption of plasma proteins other than Fg caused the high levels of activation observed for plasma preadsorbed surfaces. PMID- 11026631 TI - Surface-immobilized dextran limits cell adhesion and spreading. AB - Dextran has recently been investigated as an alternative to polyethylene glycol (PEG) for low protein-binding, cell-resistant coatings on biomaterial surfaces. Although anti-fouling properties of surface-grafted dextran and PEG are quite similar, the multivalent properties of dextran are advantageous when high-density surface immobilization of biologically active molecules to low protein-binding surface coatings is desired. The preferred methods of dextran immobilization for biomaterial applications should be simple with minimal toxicity. In this report, a method is described for covalent immobilization of dextran to material surfaces which involves low residual toxicity reagents in mild aqueous reaction conditions. 70 kDa MW dextran was immobilized on glass and polyethylene terephthalate (PET) surfaces. 3T3 fibroblast cell adhesion was compared on untreated, aminated, and dextran-coated materials. Dextran coatings effectively limited cell adhesion and spreading on glass and PET surfaces in the presence of serum-borne cell adhesion proteins. With dextran-based surface coatings, it will be possible to develop well-defined surface modifications that promote specific cell interactions and perhaps better performance in long-term biomaterial implants. PMID- 11026632 TI - Correlation of transarterial transport of various dextrans with their physicochemical properties. AB - Local vascular drug delivery provides elevated concentrations of drug in the target tissue while minimizing systemic side effects. To better characterize local pharmacokinetics we examined the arterial transport of locally applied dextran and dextran derivatives in vivo. Using a two-compartment pharmacokinetic model to correct the measured transmural flux of these compounds for systemic redistribution and elimination as delivered from a photopolymerizable hydrogel surrounding rat carotid arteries, we found that the diffusivities and the transendothelial permeabilities were strongly dependent on molecular weight and charge. For neutral dextrans, the effective diffusive resistance in the media increased with molecular weight approximately 4.1-fold between the molecular weights of 10 and 282 kDa. Similarly, endothelial resistance increased 28-fold over the same molecular weight range. The effective medial diffusive resistance was unaffected by cationic charge as such molecules moved identically to neutral compounds, but increased approximately 40% when dextrans were negatively charged. Transendothelial resistance was 20-fold lower for the cationic dextrans, and 11 fold higher for the anionic dextrans, when both were compared to neutral counterparts. These results suggest that, while low molecular weight drugs will rapidly traverse the arterial wall with the endothelium posing a minimal barrier, the reverse is true for high molecular weight agents. With these data, the deposition and distribution of locally released vasotherapeutic compounds might be predicted based upon chemical properties, such as molecular weight and charge. PMID- 11026633 TI - Nitric oxide-generating polymers reduce platelet adhesion and smooth muscle cell proliferation. AB - We have developed polymeric biomaterials capable of providing localized and sustained production of nitric oxide (NO) for the prevention of thrombosis and restenosis. In the current study, we have characterized the kinetics of NO production by these materials and investigated their efficacy in reducing platelet adhesion and smooth muscle cell proliferation in vitro. Three nitric oxide donors with different half-lives were covalently incorporated into photopolymerized polyethylene glycol hydrogels. Under physiological conditions, NO was produced by these hydrogels over periods ranging from hours to months, depending upon the polymer formulation. NO production was inhibited at acidic pH, which may be useful for storage of the materials. The NO-releasing materials successfully inhibited smooth muscle cell growth in culture. Platelet adhesion to collagen-coated surfaces was also inhibited following exposure of whole blood to NO-producing hydrogels. The effects of NO production by these hydrogels on platelet adhesion and the proliferation of smooth muscle cells suggest that these materials could reduce thrombosis and restenosis following procedures such as balloon angioplasty. PMID- 11026634 TI - Perivascular graft heparin delivery using biodegradable polymer wraps. AB - Heparin remains the gold-standard inhibitor of the processes involved in the vascular response to injury. Though this compound has profound and wide-reaching effects on vascular cells in culture and animal models, its clinical utility has been questionable at best. It is clear that the mode of heparin delivery is critical to its potential and it may well be that routine forms of administration are insufficient to observe benefit given the heparin's short half-life and complex pharmacokinetics. When ingested orally, heparin is degraded to inactive oligomer fragments while systemic administration is complicated by the need for continuous infusion and the potential for uncontrolled hemorrhage. Thus alternative heparin delivery systems have been proposed to maximize regional effects while limiting systemic toxicity. Yet, as heparin is such a potent antithrombotic compound and since existing local delivery systems lack the ability to precisely regulate release kinetics, even site-specific therapy is prone to bleeding. We now describe the design and development of a novel biodegradable system for the perivascular delivery of heparin to the blood vessel wall with well-defined release kinetics. This system consists of heparin encapsulated poly(DL lactide-co-glycolide) (pLGA) microspheres sequestered in an alginate gel. Controlled release of heparin from this heterogeneous system could be obtained over a period of 25 days in vitro. The experimental variables affecting heparin release from these matrices were investigated. Gel permeation chromatography (GPC) and scanning electron microscopy (SEM) were used to monitor the degradation process and found to correlate well with the release kinetics. Heparin-releasing gels inhibited growth of bovine vascular smooth muscle cells in tissue culture in a dose-dependent manner. Moreover, gel release controlled vascular injury in denuding and interposition vascular graft animal models of disease even when uncontrolled bleeding was evident with standard matrix-type release. This system may therefore provide an effective means of examining the effects of various compounds in the control of smooth muscle cell proliferation in accelerated arteriopathies and also shed light on the biologic nature of these processes. PMID- 11026635 TI - Complement activation by PVA as measured by ELIFA (enzyme-linked immunoflow assay) for SC5b-9. AB - Polyvinyl alcohol (PVA) coated onto polyethylene (PE) tubes exposed to human serum for 1 hour at 37 degrees C resulted in the production of 1.03 +/- 0.04 microg/cm2 of the soluble form of the terminal membrane attack complex, SC5b-9. This was approximately 20 x that produced by the polyethylene. About one quarter of this total was found associated with the surface of PVA. SC5b-9 concentrations were determined by enzyme-linked immunoflow assay (ELIFA) a variant on ELISA that involved drawing the test sample, the antibodies and the chromogenic reagent through a nitrocellulose membrane filter. ELIFA enabled analysis of protein concentrations in the presence of SDS, so that SDS (0.05%) was used to desorb adsorbed SC5b-9 prior to analysis together with SC5b-9 in the bulk to get a more complete picture of PVA-associated complement activation. PMID- 11026636 TI - Evaluation of leukocyte adhesion on polyurethanes: the effects of shear stress and blood proteins. AB - Leukocytes are a central cell type in directing host inflammatory and immune processes; thus, its response to biomaterials is extremely important in understanding material-host interaction. Blood contacting biomaterials may activate the complement cascade, thus promote leukocyte adhesion and activation to the biomaterial surface. We hypothesize that the extent of complement-mediated leukocyte activation is modulated by the material chemical formulation and the presence of fluid shear stress. Medical-grade polyurethanes with or without 4,4' butyldiene bis(6-tert-butyl-m-cresol) antioxidant additives and a rotating disk system were utilized to study cell adhesion under a well-characterized shear stress field. Radioimmunoassay and ELISA were employed to assess the extent of complement activity. The results showed that adherent leukocyte densities decreased with increasing shear stress and that leukocyte adhesion was decreased significantly further by the presence of the antioxidant in the polyurethanes. Cell adhesion under flow conditions was abolished when complement C3 protein was depleted from the test medium. An increase in complement Factor H adsorption was observed at high shear region; however, no change in the complete complement activation was observed in the presence of shear stress as indicated by the protein S-terminal complement complex level. Based on these results, oligopeptides designed from C3a, C5a, and fibronectin were grafted onto a cell nonadhesive polymer surface to probe the molecular mechanisms of leukocyte adhesion as mediated by protein-receptor complexation. The results showed that C3a-derived peptides mediated higher adherent macrophage density when compared to that mediated by C5a- and fibronectin-derived peptides. PMID- 11026637 TI - Leukocyte chemosensory migration on vascular prosthetic biomaterial is mediated by an integrin beta2 receptor chain. AB - The ability of adherent activated leukocytes to migrate on implanted prosthetic biomaterial surfaces may be an early rate-limiting step in eliminating periprosthetic infection. The goal of this study was to explore the molecular mechanism governing leukocyte migration on the implantable cardiovascular prosthetic biomaterial, expanded polytetrafluoroethylene (ePTFE), in response to stimulation by the soluble chemokine, N-formyl-methionyl-leucyl-phenylalanine (fMLP). We used a population level migration assay to study the migration of polymorphonuclear leukocytes (PMN) on ePTFE, overlaid by a gelatin/agar composite. A theoretical random walk model was applied to describe fMLP-induced PMN migration on ePTFE in terms of an objective random cell migration coefficient, mu. Our results show that following stimulation with 0-10(-7) M fMLP, the value of mu ranged from 5.43 x 10(-9) to 1.08 x 10(-7) cm2/s, with a maximum value obtained at 10(-8) M fMLP. We probed the expression levels of various beta2 integrin receptor subunits and their contribution to the migratory function of ePTFE-adherent PMN over a wide range of fMLP concentration. We found that the expression of the integrin beta-chain, CD18, was also maximized at 10( 8) M fMLP, along with only slight changes in the expression of integrin alpha chains (CD11a,b,c). We report that treatment with antibodies against either beta or combined alpha chains, but not individual alpha chains, inhibited PMN attachment to ePTFE at 10(-8) M fMLP, suggesting the likely role of combined beta2 receptor subunits in early adhesion events following stimulation. However, treatment with only anti-CD18 significantly lowered PMN migration on ePTFE (mu = 5.98 x 10(-9) cm2/s), and this degree of inhibition was much greater than that elicited by the combined treatment with antibodies recognizing all possible alpha chains. Overall, we conclude that migratory behavior of chemokinetically stimulated PMN on ePTFE is mediated by the integrin beta chain pool, and is only weakly regulated by the integrin alpha chain. PMID- 11026638 TI - Vascular cell responses to polysaccharide materials: in vitro and in vivo evaluations. AB - Chitosan has shown promise as a structural material for a number of tissue engineering applications. Similarly the glycosaminoglycans (GAGs) and their analogs have been known to exert a variety of biological activities. In this study we evaluated the potential of GAG-chitosan and dextran sulfate (DS) chitosan complex materials for controlling the proliferation of vascular endothelial (EC) and smooth muscle cells (SMC). GAG-chitosan complex membranes were generated in vitro and seeded with human ECs or SMCs for culture up to 9d. In addition, porous chitosan and GAG-chitosan complex scaffolds were implanted subcutaneously in rats to evaluate the in vivo response to these materials. The results indicated that while chitosan alone supported cell attachment and growth, GAG-chitosan materials inhibited spreading and proliferation of ECs and SMCs in vitro. In contrast, DS-chitosan surfaces supported proliferation of both cell types. In vivo, heparin-chitosan and DS-chitosan scaffolds stimulated cell proliferation and the formation of a thick layer of dense granulation tissue. In the case of heparin scaffolds the granulation layer was highly vascularized. These results indicate that the GAG-chitosan materials can be used to modulate the proliferation of vascular cells both in vitro and in vivo. PMID- 11026639 TI - Heparinized bovine pericardium as a novel cardiovascular bioprosthesis. AB - A novel chemical modification of biological tissues was developed by the direct coupling heparin to bovine pericardium (BP). The heparinization involves pretreatment of BP using GA and followed by grafting heparin to BP by the reaction of residual aldehyde and amine group of heparin. BP was modified by direct coupling of heparin and the effect of heparin coupling on calcification was evaluated in vitro and in vivo. Heparinized BP was characterized by measuring shrinkage temperature, mechanical properties, digestion resistance to collagenase enzyme, in vitro cytotoxicity, and in vivo calcification. Thermal and mechanical properties showed that the durability of heparin-treated tissue increased as compared with fresh tissue and GA-treated tissue. Resistance to collagenase digestion revealed that heparin-treated tissue has greater resistance to enzyme digestion than did fresh tissue and GA-treated tissue. Heparinized tissue had shown to be non-cytotoxic, however, relatively high cytotoxicity was observed in the GA-treated tissues due to the release of GA. In vivo calcification study demonstrated much less calcium deposition on heparin-treated BP than GA-treated one. Obtained results attest to the usefulness of heparinized BP for cardiovascular bioprostheses. PMID- 11026640 TI - Effect of angiotensin-converting enzyme inhibitors and nitroxy groups on human coronary resistance vessels in vitro. AB - We investigated the interaction between nitroxy groups and angiotensin-converting enzyme (ACE) inhibitors to assess the role of sulfhydryl groups and adenosine triphosphate (ATP)-sensitive potassium channels in vasodilation of human coronary resistance vessels in vitro. Coronary resistance vessels were resected from the right atrial appendage of 27 patients undergoing open heart surgery. The vessel ends were inserted into a microglass pipette with the internal pressure maintained at 40 mm Hg. Nitroglycerin did not change the vasoresponse, whereas nicorandil induced a concentration-dependent vasodilation that was not affected by methylene blue, but was markedly inhibited by glibenclamide. The ACE inhibitors, captopril, with a sulfhydryl group (1 x 10(-6) M), and enalaprilat, without a sulfhydryl group (1 x 10(-6) M), were added to either nitroglycerin or nicorandil to assess the incremental response of the sulfhydryl group to vasodilation. The addition of captopril or L-cysteine (1 x 10(-6) M) enhanced the activity of both nitroglycerin and nicorandil, whereas addition of enalaprilat did not. The responses of nicorandil and nitroglycerin to captopril and were similar. Cromakalim was not enhanced by L-cysteine or captopril. The response of nitroglycerin was not enhanced by captopril or L-cysteine after addition of N(G) monomethyl-L-ARGININE (L-NMMA). Both nitroglycerin and nicorandil exhibited an increase in vasodilation in the presence of an ACE inhibitor containing a sulfhydryl group. The mechanism of the vasodilatory action in the coronary resistance vessels may involve the opening of an ATP-sensitive potassium channel and subsequent guanylate cyclase activation. These interactions have important clinical implications. PMID- 11026641 TI - Water-soluble hexasulfobutyl [60] fullerene inhibits plasma lipid peroxidation by direct association with lipoproteins. AB - Free radicals can induce lipid peroxidation, leading to the formation of atherosclerosis. A new class of water-soluble C60 derivative, hexasulfobutyl [60] fullerene [C60-(CH2CH2CH2CH2SO3Na)6; (FC4S)], comprising six sulfobutyl functional groups covalently bound on a C60 cage, is a potent free radical scavenger. This study sought to define the effect of FC4S in protecting plasma from peroxidation. At concentrations of 10-100 microM, FC4S efficiently protected plasma against Cu2+-induced oxidation, as shown by maintenance of apoprotein B integrity and decrease in oxidative products levels, conjugated diene, and thiobarbituric acid-reactive substances. Addition of FC4S to both plasma and isolated lipoproteins, including very low density, low-density, and high-density lipoproteins, resulted in an increased mobility of the lipoprotein on agarose gel electrophoresis. This was attributed to FC4S associating with the lipoproteins because of the negative charge of the sulfonate groups after hydrolysis in the electrophoretic buffer. When lipoprotein was oxidized by 2,2-azobis (2,4-dimethyl valeronitrile), which produces peroxyl radicals within lipoprotein, but not in the aqueous phase, the FC4S still efficiently inhibited lipoprotein oxidation. These data substantiate that FC4S acts efficiently in protecting plasma lipid from oxidation by associating with lipoprotein to scavenge free radicals in both the aqueous and lipophilic phases. PMID- 11026642 TI - Treatment with a combined endothelin A/B-receptor antagonist does not prevent chronic renal allograft rejection in rats. AB - A markedly increased expression of endothelin (ET)-1 has been observed in renal allografts with chronic rejection, one of the most common causes of kidney graft loss. In this study we investigated the effect of treatment with a combined ET A/B-receptor antagonist on the course of chronic renal allograft rejection. Experiments were performed in the Fisher-to-Lewis rat model of chronic rejection. Lewis-to-Lewis isografts and uninephrectomized Lewis rats served as controls. Animals were treated with either the oral combined ET-A/B-receptor antagonist LU224332 (20 mg/kg/day) or vehicle. Animal survival, blood pressure, creatinine clearance, proteinuria, and urinary ET excretion were investigated for 24 weeks. Kidneys were removed for light-microscopic evaluation and immunohistochemical assessment of cell-surface markers. Treatment with LU224332 did not improve survival after 24 weeks (0.47 vs. 0.38; p > 0.05 by log-rank test), nor did it have an influence on blood pressure, creatinine clearance, or proteinuria. Combined ET-A/B-receptor blockade was associated with a reduction of expression of cell-surface markers for macrophages (EDI), T-cells (R73), and major histocompatibility complex (MHC) II (F17-23-2), but did not lead to an improvement of histologic changes of chronic allograft rejection. Our data show that blocking both ET-A- and -B receptors, in opposition to a previously published beneficial effect of selective ET-A blockade, does not prevent the progression of chronic renal allograft rejection and does not prolong survival in this model. Functional integrity of the ET-B receptor therefore seems to play an important role in the nephroprotection provided by selective ET-A-receptor antagonists in chronic renal allograft nephropathy. PMID- 11026643 TI - Adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes. AB - Vascular endothelial growth factor (VEGF) is an angiogenic growth factor that stimulates endothelial cell proliferation, increases endothelial permeability, and promotes collateral vessel formation. We transferred human VEGF gene into rat cardiac myocytes using adeno-associated virus (AAV) vectors and investigated whether VEGF secreted from the transduced cardiac myocytes promoted proliferation of endothelial cells. We produced VEGF-expressing AAV vectors (AAV-VEGF) by the adenovirus-free method. Immunoblotting revealed VEGF protein expression in AAV VEGF-transduced rat cardiac myocytes. More than 60% of cardiac myocytes were stained positively on immunohistochemical staining using anti-VEGF antibody. Concentration of VEGF in the culture medium of AAV-VEGF-transduced myocytes was increased in a vector dose-dependent manner, and VEGF secretion from the transduced myocytes persisted for > or = 14 days. Thymidine incorporation into human vascular endothelial cells was significantly increased by incubation with the conditioned medium from AAV-VEGF-transduced myocytes. This increased thymidine uptake was significantly inhibited by anti-VEGF antibody. We demonstrated here that AAV-mediated VEGF gene transfer into cardiac myocytes induces the secretion of functional VEGF. PMID- 11026644 TI - Release of nitric oxide after acute hypertension. AB - We have shown that NO production, assessed by measuring changes in plasma nitrate concentration, is down-regulated when blood pressure falls. This study intended to determine first, whether NO-derived plasma nitrate varies in response to increases in blood pressure induced by different mechanical and pharmacologic stimuli, including angiotensin II and catecholamines; and second, specifically to study the interaction between angiotensin II and NO production. An intravenous infusion (4-10 min) of norepinephrine (7.5 microg/kg/min), phenylephrine (30 microg/kg/min), or angiotensin II (0.3 and 3 microg/kg/min) caused hypertension accompanied by an increase in plasma nitrate, as assessed by high-performance capillary electrophoresis. Mechanical hypertension elicited by aortic occlusion also was accompanied by an increase in plasma nitrate. Angiotensin II (0.03, 0.3, and 3 microg/kg/min, 10 min) dose-dependently increased blood pressure. The intermediate and high dose, but not the low dose, of angiotensin II increased plasma nitrate concentration. N(G)-nitro-L-arginine methyl ester (L-NAME) lowered the basal concentration of plasma nitrate, abolished the increase in plasma nitrate elicited by angiotensin II and norepinephrine, and potentiated the pressor effect of the low dose of angiotensin II, although this dose did not increase NO production. L-NAME also potentiated the pressor effects of the intermediate dose of angiotensin II. This study demonstrates that an augmented systemic production of NO, measured as an increase in plasma nitrate, takes place after acute hypertension. The results of this study suggest that an increase in NO generation occurs when angiotensin II hypertension exceeds a certain limit, below which the basal production of NO is sufficient to compensate the vasoconstriction. PMID- 11026645 TI - Neuropeptide Y1- and Y2-receptor-mediated cardiovascular effects in the anesthetized guinea pig, rat, and rabbit. AB - Neuropeptide Y (NPY) causes vasoconstriction through Y1-receptors and inhibits vagal bradycardia through presynaptic Y2-receptors. These effects of NPY were investigated in anesthetized guinea pigs, rats, and rabbits to find the most suitable species for evaluation of Y1- and Y2-active agents in vivo. The increase in blood pressure (through Y1) of lower doses of NPY was similar in the three species (ED50, 0.9 +/- 0.13, 0.8 +/- 0.39, and 0.6 +/- 0.09 nmol/kg, respectively), but higher doses had depressor effects in four of six rats. Vagal bradycardia, induced by electrical stimulation of the right cervical vagus nerve, was inhibited by NPY in the guinea pig and in the rat (ED35, 3.5 +/- 0.46 and 11.2 +/- 1.79 nmol/kg, respectively; p < 0.05) but not in the rabbit. In the guinea pig, the Y2-receptor-preferring fragment NPY(3-36) and the selective Y1 receptor antagonist H 409/22 were used to confirm that the increase in blood pressure was mediated solely through the Y1-receptor and the vagal inhibition solely through the Y2-receptor. Aside from the cardiovascular effects, NPY caused a decrease in the body temperature and inhibited vagal bronchoconstriction in this species. Considering that NPY may cause depressor effects in the rat and has no effect on the vagal bradycardia in the rabbit, the guinea pig is preferable to both these species for assessment of Y1- and Y2-receptor-active agents in vivo. PMID- 11026646 TI - Raloxifene and estrogen inhibit neointimal thickening after balloon injury in the carotid artery of male and ovariectomized female rats. AB - The effects of raloxifene and 17alpha-ethinyl estradiol (EE2) on intimal thickening in response to balloon injury were tested in male and ovariectomized female rats. In male rats, oral raloxifene and EE2, administered either by gavage or in the diet, inhibited arterial intimal thickening in response to balloon injury to a maximum of approximately 60 and 50%, respectively. The effect of oral raloxifene to decrease cholesterol was observed at doses (> or = 3 mg/kg/day) higher than those required to inhibit intimal thickening (> or = 0.03 mg/kg/day). Coadministration of the estrogen receptor antagonist, ICI 182,780 (5 mg/kg/day, s.c.), blocked the inhibition of balloon injury by raloxifene and EE2. Direct adventitial delivery of raloxifene (0.03 mg/kg/day) and EE2 (0.001 mg/kg/day) to the vascular wall inhibited intimal thickening by 63 and 53%, respectively. In ovariectomized female rats, oral raloxifene (0.01-3.0 mg/kg/day) and EE2 (0.08 mg/kg/day) inhibited intimal thickening to a maximum of 32 and 60%, respectively. Together, these data suggest that raloxifene and EE2, inhibit balloon arterial injury in the rat through direct effects on the vascular wall that involve the estrogen receptor and are at least partially independent of serum cholesterol. PMID- 11026647 TI - Binding properties of beta-blockers at recombinant beta1-, beta2-, and beta3 adrenoceptors. AB - The human heart contains at least four distinct beta-adrenoceptor subtypes, three of which have been cloned. However, the binding properties of beta-blockers to the different beta-adrenoceptor subpopulations are not yet thoroughly characterized. Human beta1-, beta2- and beta3-adrenoceptors were expressed in COS 7 cells and [125I]iodocyanopindolol saturation binding, and competition experiments with commonly used beta-blockers were performed in the respective membrane preparations. Atenolol and metoprolol were about fivefold selective for beta1- versus beta2- and beta3-adrenoceptors. Bisoprolol was approximately 15 fold selective for beta1- versus beta2- and approximately 31-fold selective for beta1- versus beta3-adrenoceptors. Carvedilol was nonselective for any beta adrenoceptor subtype. We conclude that the beta1-selectivities of atenolol, metoprolol, and bisoprolol are lower in COS cell membranes compared with previous investigations performed in native membranes. All beta-blockers investigated bind to beta3-adrenoceptors. Differential binding properties to beta3-adrenoceptors might imply different responses as to body weight, cardiac contractility, heart rate, and growth regulation. This might imply differential indications for the drugs investigated. PMID- 11026648 TI - Effects of ACE inhibitor, AT1 antagonist, and combined treatment in mice with heart failure. AB - We tested the hypothesis that a combination of angiotensin-converting enzyme inhibitor (ACEi) and angiotensin II type 1 receptor antagonist (AT1-ant) may have an additive cardioprotective effect in mice with heart failure (HF), because these two agents could have other mechanisms of action besides interrupting the renin-angiotensin system. ACEi prevent degradation of bradykinin. During treatment with AT1-ant, increased angiotensin II could activate AT2 receptors, with an antitrophic effect. To test this hypothesis, we used a mouse model of HF induced by myocardial infarction. Seven days after surgery, mice were divided into six groups and treated for 23 weeks: (a) sham ligation; (b) HF-vehicle; (c) HF-ACEi; (d) HF-AT1-ant; (e) HF-ACEi + AT1-ant (half dose of each); and (f) HF ACEi + AT1-ant (full dose of each). Cardiac function was evaluated in conscious mice during the treatment period. The HF-vehicle group showed significantly decreased left ventricular (LV) ejection fraction (EF), shortening fraction (SF), and cardiac output (CO) and increased LV dimensions, interstitial collagen, and myocyte cross-sectional area (MCSA) compared with controls. Treatment with ACEi or AT1-ant significantly increased EF, SF, and CO and decreased LV dimensions and MCSA in mice with HF. However, a combination of these drugs did not improve cardiac function more than ACEi or AT1-ant alone. We concluded that ACEi and AT1 ant have similar cardioprotective effects and may reach maximal effect when given individually; thus no further improvement can be achieved with combined therapy in mice with HF. PMID- 11026649 TI - Effects of indapamide in rats with pressure overload left ventricular hypertrophy. AB - We studied the in vivo effects of the antihypertensive diuretic agent indapamide on left ventricular (LV) morphology in chronically pressure-overloaded rat hearts. LV pressure and subsequently LV mass were increased by banding the ascending aorta over a period of 6 weeks. Thereafter, animals were treated with low-dose (1 mg/kg/day, n = 9) or high-dose (10 mg/kg/day, n = 9) indapamide for another 6 weeks. Low-dose indapamide treatment reduced LV weights as compared with vehicle-treated controls (n = 9; -12%; p = 0.008). Furthermore, low-dose indapamide treatment resulted in a decrease of myocyte volume (59.0 +/- 10.6 vs. 79.0 +/- 9.8 m3 x 10(-27); p < 0.05) and an improvement of molecular markers of hypertrophy: a reduction of LV atrial natriuretic factor mRNA expression (-37%; p < 0.05), and an increase of the V1/V3 myosin ratio (+121%; p < 0.05). Low-dose indapamide also reduced significantly plasma (-65%) and LV angiotensin-converting enzyme (ACE) activities (-74%) as well as LV mRNA levels (-24%). These changes were observed despite continued pressure overload of the LV and despite a lack of significant changes in sodium excretion with the prolonged administration of low dose indapamide. High-dose indapamide treatment showed no significant effects on LV mass, structure, and gene expression. Furthermore, high-dose indapamide increased plasma renin activity substantially, whereas low-dose treatment was without effect on circulating renin. In conclusion, in rats with continuous LV pressure-overload low-dose treatment with indapamide leads to mild regression of cardiac hypertrophy, accompanied by a downregulation of components of the cardiac renin-angiotensin system. These effects may be mediated by mechanisms apart from the known diuretic and antihypertensive actions of indapamide, because sodium excretion and blood pressure were stable with long-term treatment and are unlikely to be related to LVH regression in this model. PMID- 11026650 TI - Effects of tumor necrosis factor-alpha on basal and stimulated endothelium dependent vasomotion in human resistance vessel. AB - The aim of this study was to determine whether tumor necrosis factor (TNF)-alpha would impair basal and stimulated endothelium-dependent vasomotion in human resistance vessel. Changes in baseline and acetylcholine (ACh)-induced forearm vascular resistance (FVR) were measured plethysmographically before and after a low-dose intraarterial forearm infusion of TNF-alpha according to the following three protocols in healthy volunteers. In the condition without pretreatment, basal FVR was significantly increased by TNF-alpha (from 30.5 +/- 4.8 to 39.9 +/- 5.9 units; p < 0.01), whereas ACh-induced minimal FVR did not differ between pre- and post-TNF-alpha states. In the condition after pretreatment with the cyclooxygenase inhibitor acetylsalicylic acid, although the vascular effects of TNF-alpha on basal FVR appeared to be blocked (37.1 +/- 5.3 vs. 37.6 +/- 5.2; NS), ACh-induced minimal FVR did not differ between pre- and post-TNF-alpha states. In the condition after pretreatment with the nitric oxide (NO) synthase inhibitor N(G)-monomethyl-L-arginine, the vascular effect of TNF-alpha on basal FVR was diminished, and the ACh-induced maximal dilatory response was significantly blunted after TNF-alpha compared with before TNF-alpha (minimal FVR: 30.4 +/- 12.0 vs. 12.3 +/- 4.2 units; p < 0.05). These findings suggest that brief exposure of the human forearm resistance artery to TNF-alpha may increase basal bioavailability of the vasoconstrictor prostaglandin and reduce basal bioavailability of NO. In the stimulated condition, TNF-alpha-induced vascular dysfunction may be overwhelmed by increased NO bioavailability in healthy humans. PMID- 11026651 TI - Regulation of type-2 corticotropin-releasing hormone receptor mRNA in rat heart by glucocorticoids and urocortin. AB - A novel subtype of corticotropin-releasing hormone (CRH) receptor, designated type-2 CRH receptor (CRHR-2), has been cloned by a number of laboratories, and its mRNA has been found to be distributed not only in the brain but in peripheral tissues such as heart and skeletal muscle. To date, however, the regulation of CRHR-2 mRNA is poorly understood. Therefore, we examined the effect of glucocorticoid treatment, adrenalectomy, and systemic administration of urocortin, a possible endogenous ligand for CRHR-2, on heart CRHR-2 mRNA levels in male Wistar rats, using in situ hybridization histochemistry. CRHR-2 mRNA in the heart was significantly decreased 9 h after systemic administration of urocortin (5 microg/kg b.w.). Systemic administration of corticosterone (CORT; 10 mg/rat/day for 12 days) or CORT pellet (200 mg) implant for 7 and 14 days also decreased CRHR-2 mRNA in the heart, whereas it was unchanged 7 days after adrenalectomy. Thus, similar regulation of CRHR-2 mRNA in the rat heart by its ligand and glucocorticoids was observed. The precise mechanism of the regulation of CRHR-2 mRNA in the heart and the physiologic significance of cardiac CRHR-2 remains to be elucidated. PMID- 11026652 TI - Inhibition of vascular endothelial growth factor activity by transfection with the soluble FLT-1 gene. AB - Vascular endothelial growth factor (VEGF)-induced angiogenesis is involved in the etiology of some cardiovascular diseases. The soluble form of VEGF receptor, FLT 1 (sFLT-1), is a potent antagonist of VEGF. Therefore, we investigated whether transfection with the sFLT-1 gene could inhibit VEGF-induced angiogenesis. Human embryonic kidney (HEK)-293 cells were transfected with plasmids containing VEGF and sFLT-1 (pCMV-VEGF and pCMV-sFLT-1) by the calcium-phosphate co-precipitation method. VEGF- and/or sFLT-1-transfected HEK-293 cells were incubated for 24 h, and then conditioned medium was collected. The effects of conditioned medium on angiogenesis were tested by incorporation of [3H]thymidine into human umbilical vein endothelial cells (HUVECs). Expression of VEGF protein was determined by Western blotting. The conditioned medium from sFLT-1 gene-transfected HEK-293 cells significantly inhibited recombinant VEGF-induced increase in [3H]thymidine incorporation by HUVECs. VEGF gene-transfected HEK-293 cells secreted VEGF protein into conditioned medium. This conditioned medium increased [3H]thymidine incorporation by HUVECs, which was significantly inhibited by co-transfection of sFLT-1 gene with VEGF gene. These observations suggested that sFLT-1 gene transfer could inhibit VEGF-induced DNA synthesis of vascular endothelial cells. PMID- 11026653 TI - Thiol repletion prevents venous thrombosis in rats by nitric oxide/prostacyclin dependent mechanism: relation to the antithrombotic action of captopril. AB - Clinical and experimental data have recently accumulated for antithrombotic action of angiotensin-converting enzyme inhibitors (ACE-1s). We have shown previously that captopril (which contains a thiol group in the moiety) exerts more pronounced antithrombotic activity than does an equipotent dose of enalapril (the drug devoid of the thiol group). To clarify the relative importance of the presence of the thiol group in the molecule versus angiotensin-converting enzyme (ACE) inhibitory properties in the antithrombotic action of captopril, rats were treated with captopril (5 mg/kg twice daily; CAP), epicaptopril (stereoisomer of captopril devoid of ACE-inhibitory properties; 5 mg/kg twice daily; EPI), N acetylcysteine (3.75 mg/kg twice daily; ACC), enalapril (3 mg/kg once daily; ENA), or distilled water (VEH) for 10 days, per os. After ligation of the vena cava, the incidence of the venous thrombosis and/or the thrombus weight decreased significantly in all but the ENA-treated groups when compared with control rats. The effect of CAP, EPI, and ACC was accompanied by a marked reduction of euglobulin clot lysis time and, with the exception of ACC, by an increase in prothrombin time in the blood collected from the site of the thrombus formation. Antithrombotic activity of EPI was completely abolished by nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) or indomethacin, with the parallel reversal of fibrinolytic and coagulation parameters toward normal. Activated partial thromboplastin time, mean blood pressure, and bleeding time were not altered by either of the administered drugs. Thus, we demonstrated that thiol compounds exert antithrombotic activity by increasing fibrinolysis and/or suppression of the extrinsic pathway of the coagulation cascade in a nitric oxide/prostacyclin-dependent manner. PMID- 11026654 TI - QT-prolonging effects of sparfloxacin, a fluoroquinolone antibiotic, assessed in the in vivo canine model with monophasic action potential monitoring. AB - Sparfloxacin, a fluoroquinolone antibacterial agent, prolongs cardiac repolarization, which may predispose to torsades de pointes. This study was designed to assess simultaneously the hemodynamic and electrophysiologic effects of sparfloxacin using the halothane-anesthetized, closed-chest in vivo canine model (n = 6). Sparfloxacin was intravenously administered in the following two doses with a pause of 20 min, a clinically relevant dose of 3.0 mg/kg/10 min and a 10 times higher dose of 30 mg/kg/10 min. After the low dose of sparfloxacin, cardiac output increased, heart rate decreased, and ventricular repolarization and refractory periods were prolonged. After the high dose, cardiac output increased, whereas heart rate and mean blood pressure decreased, and ventricular repolarization and effective refractory periods were prolonged. The increment was greater in repolarization than in refractoriness, indicating an increase of electrical vulnerability. Because sparfloxacin prolonged repolarization in a reverse use-dependent manner, its negative chronotropic effect may have potentiated the QT prolongation. Left ventricle preload, left ventricular contraction, and AV nodal as well as intraventricular conduction were minimally affected. These results suggest that caution should be used when administering sparfloxacin to patients having risk factors for QT prolongation. PMID- 11026655 TI - In vivo characterization of the novel neuropeptide Y Y1 receptor antagonist H 409/22. AB - We studied the effects of the novel neuropeptide Y (NPY) Y1 receptor antagonist H 409/22, and its inactive enantiomer H 510/45, on vascular responses evoked by endogenous and exogenous NPY in the pig in vivo. H 409/22 and H 510/45 were given as 30-min infusions, and the antagonistic effects and circulating plasma concentrations were measured. The initial and terminal half-lives of H 409/22 in plasma were approximately 3 and 30 min, respectively. In pigs pretreated with reserpine and transection of sympathetic nerves (depletion of noradrenaline), sympathetic nerve stimulation evoked nonadrenergic vasoconstrictor responses in kidney and hindlimb, mediated by neuronally released NPY. Significant inhibition of these vasoconstrictor responses, as well as of vascular responses to injections of exogenous NPY, were seen during a low-dose infusion of H 409/22 (1.8 nmol/kg/min), when plasma levels of the antagonist reached 77 +/- 8 nM. Greatest inhibitory effects were seen at the highest dose of H 409/22 (180 nmol/kg/min, giving plasma levels of 7.4 +/- 0.6 microM) when all vascular responses evoked by NPY were strongly attenuated or largely abolished. H 510/45 did not affect any of the vascular responses studied. It is concluded that H 409/22 potently and dose-dependently antagonizes vascular responses to exogenous and endogenous NPY in the pig, and thus represents an interesting tool for studies on NPY Y1 receptor-mediated effects in vivo. PMID- 11026656 TI - Adrenomedullin attenuates pressor response to angiotensin II in conscious sheep. AB - Biologic actions attributed to adrenomedullin include reduction of arterial pressure and suppression of aldosterone secretion. To assess possible in vivo antiangiotensin II actions of adrenomedullin, we examined hemodynamic and adrenal responses to stepped angiotensin II infusions with or without co-infusions of adrenomedullin (33 ng/kg/min) in conscious sheep under controlled conditions of a low sodium intake. Plasma adrenomedullin levels rose during peptide infusions (p < 0.001) to plateau at approximately 15-18 pM. The dose-dependent pressor response (15-20 mm Hg) of angiotensin II was both delayed and markedly attenuated (p = 0.017) by adrenomedullin, which also stimulated heart rate (p < 0.001) and cardiac output (p < 0.001). Adrenomedullin prevented the angiotensin II-induced increase in peripheral resistance (p < 0.001). Plasma aldosterone responses to angiotensin II were variable and were not significantly altered by concomitant adrenomedullin infusion. In conclusion, low-dose infusion of adrenomedullin administered to conscious sheep on a low-salt diet clearly antagonized the vasopressor actions of administered angiotensin II while stimulating cardiac output and heart rate. The data suggest a possible role for adrenomedullin in cardiovascular homeostasis in part through antagonism of the vasopressor action of angiotensin II. PMID- 11026657 TI - The effect of chronic azithromycin therapy on soluble endothelium-derived adhesion molecules in patients with coronary artery disease. AB - In patients with coronary artery disease (CAD), azithromycin therapy is associated with decreased cytokine levels and overall reduction of inflammation. Chlamydia pneumoniae (C.Pn) is a common pathogen that may be an important factor in the development and progression of atherosclerosis. Cell-adhesion molecules have an important role in recruitment of inflammatory cells during plaque development and are expressed by endothelial cells on activation. We sought to define the effect of treatment with azithromycin on circulating levels of soluble vascular cell-adhesion molecule (VCAM-I), intercellular adhesion molecule (ICAM 1), and E-selectin in patients with CAD. Plasma concentrations of VCAM-1, ICAM-1, and E-selectin were measured in 40 patients with documented CAD and a positive (> or = 1:16) immunoglobulin G (IgG) titer against C.Pn, 20 subjects with normal coronary arteries, and 14 healthy volunteers. Patients were assigned randomly to receive either 500 mg/wk of azithromycin or placebo for 3 months. Serum samples were obtained at baseline, at 3 months, and during the follow-up visit at 6 months. Patients with documented CAD exhibited elevation of VCAM-1 (535 +/- 227 ng/ ml; p = 0.0001) and E-selectin (69 +/- 29 ng/ml; p = 0.006), but not ICAM-1 (321 +/- 65 ng/ml) concentrations as compared with the patients with angiographically proven normal coronary arteries (252 +/- 80; 50 +/- 22; and 311 +/- 40 ng/ml) and healthy controls (110 +/- 18; 29 +/- 2; and 238 +/- 47 ng/ml, respectively). Prolonged treatment with azithromycin did not significantly affect the plasma levels of soluble VCAM-1, ICAM-1, and E-selectin. Soluble markers of endothelial activation are markedly increased in patients with documented CAD as compared with those with normal coronary arteries and healthy controls. Despite substantial heterogeneity in plasma E-selectin, ICAM-1, and VCAM-1 levels, long term azithromycin treatment did not affect plasma levels of these adhesion molecules, indicative of endothelial activation, over a period of 6 months. PMID- 11026658 TI - Microsatellites: consensus and controversy. AB - Microsatellite DNA loci have recently been adopted for many biological applications. Comparative studies across a wide range of species has revealed many details of their mutational properties and evolutionary life cycles. Experience shows that a full understanding of these processes is essential to ensure the effective use of microsatellites as analytical tools. In this article, we review the controversies that have arisen as biologists have taken up this new technology and the emerging consensus that has resulted from their debates. We point to the need for comparative DNA sequencing studies to produce input data for a new generation of theoretical models of microsatellite behaviour. We conclude by presenting our own conceptual model, 'Snakes and Ladders', as an aid to theory development. PMID- 11026659 TI - Lipids and fatty acids of two pelagic cottoid fishes (Comephorus spp) endemic to Lake Baikal. AB - Matured females of two Lake Baikal endemic fish species, Comephorus baicalensis and Comephorus dybowski, have been investigated for lipid of the whole body and specific tissues (liver, muscles, ovaries), phospholipid classes and fatty acids of neutral and polar lipids. Total lipid in the body (38.9% fresh weight), liver (23.5%) and muscles (14.5%) of C. baicalensis were greater than those of C. dybowski (4.7, 8.7 and 2.6%, respectively); only their ovaries were similar (5.3 and 5.6% lipid, respectively). In both species, phosphatidylcholine and phosphatidylethanolamine were the major phospholipids, ranging from 60.7 to 75.1% of total phospholipid and 14.5-25.7%, respectively. In most cases, monounsaturated fatty acids (MUFA) were the major fatty acid group in C. baicalensis, whereas polyunsaturated fatty acids (PUFA) were the major group in C. dybowski. The MUFA 18:1(n-9) prevailed over other fatty acids in C. baicalensis and varied from 19% in polar lipids of muscles to 56.1% in neutral lipids of muscles. In polar lipid of C. dybowski, the PUFA 22:6(n-3) prevailed over other fatty acids in muscles and ovaries, while 16:0 dominated polar liver lipids and neutral lipids of all tissues. Other major fatty acids included 16:1(n 7), 18:1(n-7), and 20:5(n-3). Values of the (n-3)/(n-6) fatty acid ratio for neutral lipids of C. baicalensis (0.5-0.9) are well below the range of values characteristic either for marine or freshwater fish, while these values for polar lipids (1.6-1.8) are in the range typical of freshwater fish. Neutral lipid fatty acid ratios in C. dybowski (2.5-3.1) allow it to be assigned to freshwater fish, but polar lipids (2.8-3.7) leave it intermediary between freshwater and marine fish. PMID- 11026660 TI - Purification and partial characterisation of tentatively classified acid phosphatase from the earthworm Eisenia veneta. AB - In order to use leakage of lysosomal acid phosphatase (AP) as a biomarker of stress to earthworms, more information about AP's in earthworms are needed. This paper describes the details about tentatively classified APs in the earthworm Eisenia veneta. Two isoenzymes (enzyme I and II) of acid phosphatase (AP) and one alkaline phosphatase (enzyme III) from the earthworm E. veneta were separated by gel filtration. All three enzymes were further purified and concentrated on a Con A Sepharose 4B column. Enzyme I was inhibited by tartrate, showed an optimal pH range between 4.0 and 5.0 and was assumed to be of lysosomal origin. Enzyme II was the major enzyme showing the highest activity of the three enzymes. It was expected to be a lysosomal AP under physiological conditions. Enzyme II had a molecular mass 113 kDa and was composed of apparently identical polypeptide chains of 36 kDa each. This enzyme was inhibited by tartrate, showed an optimal pH in the range 6.0-7.5 and was slowly degraded at temperatures above 40 degrees C. Enzyme III is not inhibited by tartrate and has a pH-optimum > 9. The subcellular location under physiological conditions was assumed to be the cytosol. PMID- 11026661 TI - Regulation of the futile cycle of fructose phosphate in sea mussel. AB - Carbohydrate metabolism in mussels shows two phases separated seasonally. During summer and linked to food supply, carbohydrates, mainly glycogen, are accumulated in the mantle tissue. During winter, mantle glycogen decreases concomitantly with an increase in triglyceride synthesis. In spring, after spawning, the animals go in to metabolic rest until the beginning of a new cycle. This cycle is regulated by the futile cycle of fructose phosphate that implicates PFK-1 and FBPase-1 activities. These enzymes and the bifunctional PFK-2/FBPase-2 that regulates the Fru-2,6-P2 levels, are seasonally modulated by covalent phosphorylation/dephosphorylation mechanisms, as a response to unknown factors. The futile cycle of the fructose phosphates also controls the transition from physiological aerobiosis to hypoxia. The process is independent of the phosphorylation state. In this sense, a pH decrease triggers a small Pasteur effect during the first 24 h of aerial exposure. Variations in the concentration of Fru-2,6-P2 and AMP are the sole factor responsible for this effect. Longer periods of hypoxia induce a metabolic depression characterized by a decrease in Fru-2,6-P2 which is hydrolyzed by drop in the pH. In this review, the authors speculate on the two regulation processes. PMID- 11026662 TI - Isozyme distribution of ten enzymes and their loci in South American lungfish, Lepidosiren paradoxa (Osteichthyes, Dipnoi). AB - Scarce bibliographical data exists on the enzymes in Lepidosiren paradoxa and analysis of several enzymes was considered worthy of investigation. Distribution of ADH, ALP, FBALD, GAPDH, G3PDH, G6PDH, GPI, LDH, MDH, and PGM was identified in ten tissues (retina, heart, muscle, liver, kidney, lung, gut, gills, brain, and ovary) of the South American lungfish and compared with patterns previously described in other vertebrates. Compared with earlier results differences in the number of loci expressed were observed for ADH, G3PDH, GPI, and MDH. The number of loci expressed and/or in tissue specificity of several enzymes (ADH, FBALD, GAPDH, G3PDH, G6PDH and PGM) were found to be similar to those of other vertebrates. Differences were detected in ALP due to the absence of an intestinal specific form typical of fish, amphibians, reptiles and birds; further differences were observed in GPI and MDH due to their tissue expression. The differences in LDH involve the LDH-A4 isozyme which was most common in tissues. Overall, comparison with other vertebrates reveals that in L. paradoxa the tissue restricted expressions of some enzymes are similar, while others have retained an ancestral pattern and exhibit a more widespread tissue expression of genes. PMID- 11026663 TI - Molecular cloning and expression of an otolith matrix protein cDNA from the rainbow trout, Oncorhynchus mykiss. AB - The fish otolith is a hard tissue consisting of calcium carbonate and organic matrices. The matrix proteins play important roles in otolith formation, but little is known about the nature of these proteins. In this study, matrix proteins were extracted from the otoliths of rainbow trout, Oncorhynchus mykiss, and chum salmon, Oncorhynchus keta. EDTA-soluble matrix proteins were separated by SDS-PAGE, revealing two major components in the otoliths of both species with apparent molecular masses of 55 and 43 kDa. N-terminal and some internal amino acid sequences of the 55-kDa otolith matrix protein were determined. A cDNA fragment encoding this protein of O. mykiss was amplified by reverse transcription PCR using two degenerate primers designed from the amino acid sequences. A cDNA encoding this protein was obtained by screening a saccular cDNA library using the amplified cDNA fragment as a probe. Nucleotide sequence analysis revealed that the cDNA clone has a sequence of 2.5 kb and the open reading frame encoding 344 amino acid residues. Northern blot analysis showed that mRNA of this protein is expressed specifically in the sacculus, and consistently during the day. PMID- 11026664 TI - Detection and basic properties of carbamoyl phosphate synthetase III during teleost ontogeny: a case study in the Atlantic halibut (Hippoglossus hippoglossus L.). AB - The presence of carbamoyl phosphate synthetase III (CPSase III), catalyzing the first step of the urea cycle in fish, in Atlantic halibut (Hippoglossus hippoglossus L.) yolk-sac larvae and adult white muscle has been established using gel filtration chromatography to separate the CPSase III from the pyrimidine-pathway related CPSase II. The results are consistent with the hypothesis that teleostean fish express urea cycle enzymes during early development and with recent observations of low levels of CPSase III in muscle tissue. The presence of CPSase III in crude extracts could not be established using sensitive assay conditions to discriminate between CPSase III and CPSase II. However, kinetic characterization after chromatographic separation identified each as typical CPSase II and CPSase III activities, respectively. The CPSase III was less sensitive to activation by N-acetyl-L-glutamate and had a higher Km for ammonia than CPSase III found in other species. These results suggest that precise quantitation of low levels of CPSase III in the presence of CPSase II by assaying crude extracts may be difficult unless the enzymes are first separated and the kinetic properties of CPSase III are determined; the results indicate that assaying larval extracts of Atlantic halibut in the presence of uridine triphosphate results in CPSase activity that reflects mostly CPSase III and can, therefore, be used to measure changes in CPSase III activity. PMID- 11026665 TI - High pressure and glycolytic flux in the freshwater Chinese crab, Eriocheir sinensis. AB - The hexose part of glycolysis has been studied in the freshwater Chinese crab Eriocheir sinensis exposed to high pressure (101 ATA, i.e. 1000 m depth) at 14 degrees C and in normoxic conditions. Glycolytic fluxes (from glucose, JA and from Glucose 6 Phosphate, JB) have been determined using NADH depletion during the conversion of dihydroxy acetone phosphate into alpha-glycerol phosphate. Measurements have been performed at 14 and 19 degrees C. Pressure exposure induces an increase of glycolytic flux and a decrease of the time needed for the transition from aerobic to anaerobic glycolysis. As a consequence pressure exposed crabs have a higher potential to increase glycolytic flux than control animals at atmospheric pressure. It is concluded that high pressure known to alter numerous enzymes individually, can also modify an overall metabolic pathway. PMID- 11026666 TI - Hormone sensitive lipase mRNA in both monocyte and macrophage forms of the human THP-1 cell line. AB - The identity of the neutral cholesteryl ester hydrolase (CEH) in human monocyte/macrophages is uncertain. Prior studies indicate that hormone sensitive lipase (HSL) is a major CEH in mouse macrophages, and that HSL mRNA is present in human THP-1 monocytes. In the present study, HSL mRNA expression was examined in THP-1 cells as a function of differentiation status and cholesterol enrichment. By RT-PCR with primer pairs that span exon boundaries, HSL mRNA was demonstrated in THP-1 monocytes and phorbol-ester differentiated THP-1 macrophages. cDNA identities were confirmed by sequencing. By Northern blotting, with HSL cDNA as probe, THP-1 monocytes were found to contain HSL mRNA of approximately 3 and 3.9 kb. In THP-1 macrophages, the 3 kb mRNA was greatly diminished, while the level of the 3.9 kb mRNA was maintained. mRNA of approximately 3 and 3.9 kb are those expected of the 86-kDa (adipocyte) and 117-kDa (testicular) HSL isoforms, respectively. The presence of the testicular isoform mRNA was confirmed in THP-1 cells by amplification and sequencing of an isoform-specific cDNA. Additionally, Northern-blot comparisons showed that the 3 and 3.9 kb mRNA in THP-1 comigrated with the HSL mRNA in 3T3-L1 adipocytes and rat testis, respectively. The level of the 3.9 kb mRNA did not vary greatly with cholesterol enrichment. Thus, the HSL gene is transcribed in THP-1 cells both before and after differentiation into macrophages; after differentiation, the predominant mRNA is that for the 117-kDa isoform. This isoform is a CEH, and may mediate some CE turnover in THP-1 cells. PMID- 11026667 TI - Glycosylated salivary alpha-amylases are capable of maltotriose hydrolysis and glucose formation. AB - The physiological and/or clinical significance of sugar chains in human salivary alpha-amylase was investigated in terms of substrate-specificity for synthesized malto-oligosaccharides. Glycosylated and non-glycosylated alpha-amylases were prepared on a Sephacryl S-200 column, in which the amylases were separated into four fractions from the different affinities for Sephacryl: fraction I, amylases bearing sugar chains with sialic acid; fraction II, amylases bearing sugar chains without sialic acid; fractions III and IV, non-glycosylated amylases. These were classified according to the differences in their affinities for lectins, molecular sizes and isoelectric points. The inhibitory effect of maltotriose (G3) on starch hydrolysis of the amylase fraction, suggests that starch and G3 can be the substrate for glycosylated amylase, and that the glycosylated amylases are capable of G3 hydrolysis for conversion into maltose and glucose. Using malto oligosaccharides, G3, G4, G5 and G7, as substrates, the substrate-specificities and G3/G5 ratio of amylase activities in the four fractions were examined. Maltopentaose, G5, is routinely used as a substrate for alpha-amylase, and then we assumed that both glycosylated and non-glycosylated amylases react with G5. Moreover, the results indicate that the glycosylated amylases clearly had a higher capacity for G3 hydrolysis than the non-glycosylated amylases, although no substrate preference of either type of amylase was observed among G4, G5 and G7. Glycosylated amylases have the capacity for glucose formation from malto oligosaccharides. PMID- 11026668 TI - Enzymatic properties of sialidase from the ovary of the starfish, Asterina pectinifera. AB - A sialidase [EC 3.2.1 18] was isolated and highly purified from the ovary of the starfish, Asterina pectinifera, and its enzymatic properties were compared with those of human placental sialidase. The final preparation gave one broad protein band corresponding to sialidase activity on polyacrylamide gel electrophoresis. The molecular weight of the enzyme was 360000 by HPLC on Sigma Chrome GFC-1300 and Sephadex G-150 column chromatography, and 55000 by SDS-PAGE, suggesting the presence of a hexamer in the native protein. The optimum pH was between 3.0 and 4.0, and the enzyme liberated sialyl residues from the following compounds: alpha(2-3) and alpha(2-6) sialyllactose, colominic acid, fetuin, transferrin, gangliosides GM3, GD1a and GD1b. The enzyme was strongly inhibited by 4 aminophenyl and methyl thio-glycosides of sialic acid, but not by those glycosides of 5-amino sialic acid or sialic acid methyl ester. The enzyme was also highly inhibited by sulfated glucan and glycosaminoglycans. The substrate specificity and the effects of inhibitors on starfish sialidase were very similar to those of human placental sialidase. PMID- 11026669 TI - Hemoglobin-sulfhydryls from tortoise (Geochelone carbonaria) can reduce oxidative damage induced by organic hydroperoxide in erythrocyte membrane. AB - Sulfhydryl groups are important to avoid oxidative damage to the cell. In RBC, tert-butyl hydroperoxide (tert-BOOH) and hydrogen peroxide (H2O2) are capable of oxidizing heme and promoting lipid peroxidation. H2O2 caused greater oxidation of heme than tert-BOOH, although the oxidation of sulfhydryl groups was similar. Geochelone carbonaria Hb, a rich sulfhydryl protein, inhibited the TBA-reactive substances formation of human erythrocytes exposed to tert-BOOH by about 30%; this decrease was smaller with Geochelone denticulata Hb. Sulfhydryl reagents diminished the number of reactive sulfhydryl groups in the G. carbonaria Hb resulting in a decrease of its antioxidant power, suggesting the involvement of sulfhydryls of Hb in the protection against lipid peroxidation. PMID- 11026670 TI - cDNA cloning and sequencing of phospholipase A2 from the pyloric ceca of the starfish Asterina pectinifera. AB - Three cDNA from the pyloric ceca of the starfish Asterina pectinifera, (namely, cDNA 1, 2, and 3), encoding phospholipase A2 (PLA2), were isolated and sequenced. These cDNAs were composed of 415 bp with an open reading frame of 414 bp at nucleotide positions 1-414, which encodes 138 amino acids including N-terminal Met derived from the PCR primer. The amino acid sequence deduced from the cDNA 1 was completely consistent with the sequence determined with the starfish PLA2 protein, while those deduced from cDNA 2 and cDNA 3 differed at one and twelve amino acid residual positions, respectively, from the sequence of the PLA2 protein, suggesting the presence of multiple forms in the starfish PLA2. All of the sequences deduced from cDNA 1, 2, and 3 required two amino acid deletions in pancreatic loop region, and sixteen insertions and three deletions in beta-wing region when aligned with the sequence of mammalian pancreatic PLA2. In phylogenetic tree, the starfish PLA2 should be classified into an independent group, but hardly to the established groups IA and IB. The characteristic structure in the pancreatic loop and beta-wing regions may account for the specific properties of the starfish PLA2, e.g. the higher activity and characteristic substrate specificity compared with commercially available PLA2 from porcine pancreas. PMID- 11026671 TI - Analysis of extracellular proteins of two Perkinsus spp. isolated from the softshell clam Mya arenaria in vitro. AB - Biochemical characterization of the extracellular proteins (ECP) of two softshell clam Perkinsus spp. cloned isolates, Perkinsus chesapeaki isolate G-117 and Perkinsus marinus H-49, was performed and compared to that of the oyster-derived P. marinus isolate P-1. G-117 and H-49 demonstrated distinct differences in enzyme activities; however, all three isolates shared common bands. Substrate impregnated gels showed H-49 to possess proteolytic activities while G-117 did not. Inhibition studies revealed that H-49 ECP contain serine proteases similar to those described for P-1. The G-117 ECP lacked proteolytic activity but showed a higher production of lipolytic enzymes than H-49 or P-1. Optimal in vitro growth temperatures for the two clam isolates were generally lower than those for P-1. G-117 showed faster growth at lower salinities than either H-49 or P-1. Clam Perkinsus spp. isolates appear to be better adapted to lower salinities and temperatures than the P. murinus isolate of the eastern oyster. PMID- 11026672 TI - Purification and properties of alanine racemase from crayfish Procambarus clarkii. AB - Fresh water crayfish Procambarus clarkii is known to accumulate D-alanine remarkably in muscle after seawater acclimation, accompanied by an increase in alanine racemase activity. We have purified alanine racemase from crayfish muscle to homogeneity. The enzyme is a monomeric protein with a molecular mass of 58 kDa. It is highly specific to alanine and does not racemize L-serine, L aspartate, L-glutamate, L-valine and L-arginine. The enzyme shows the highest activity at pH 9.0 in the conversion of L- to D-alanine and at pH 8.5 in the reverse conversion. Properties such as amino acid sequence, quaternary structure, pyridoxal 5'-phosphate (PLP)-dependency, pH-dependency and kinetic parameters seem to be distinct from those of the microbial alanine racemases. Various salts including NaCl at concentrations around seawater level were potently inhibitory for the activity in both of L- to -D and D- to -L direction. PMID- 11026673 TI - Phylogenetic distribution of cysteine proteinases in beetles: evidence for an evolutionary shift to an alkaline digestive strategy in Cerambycidae. AB - We characterized the digestive proteinases of eight species of beetles to improve our understanding of the phylogenetic distribution of serine and cysteine proteinases. Serine proteinases function optimally under alkaline pH conditions, whereas cysteine proteinases require acidic pH. The phylogenetic distribution of cysteine proteinases suggests that they first appeared in an early cucujiform ancestor, however, data for some groups is patchy, and there has been speculation that they have been lost in at least one group, the long-horned beetles (Cerambycidae). The pattern we found supports the hypothesized origin of the proteinases and extends their distribution to an additional superfamily. In addition, we confirmed the presence of cysteine proteinases in some Curculionoidea. Cysteine proteinases were absent, however, from all three species of cerambycids surveyed, supporting the hypothesis that this group has reverted to the more ancestral serine (alkaline) digestive strategy. In four species we compared the pH optima for total proteolytic activity to the actual pH of the midgut and found the match between optimal and actual pH to be weaker in the cerambycids. These findings suggest that either a close correlation between midgut pH and the proteolytic pH optimum is not needed for adequate digestive efficiency, or that midgut pH is a more constrained digestive feature and there has been insufficient time for it to shift upwards to maximize serine proteinase activity. PMID- 11026674 TI - Henryk (Heini) Eisenberg. PMID- 11026675 TI - Gamma S-crystallin of bovine and human eye lens: solution structure, stability and folding of the intact two-domain protein and its separate domains. AB - Human and bovine gammaS-crystallin (HgammaS and BgammaS) and their isolated N- and C-terminal domains were cloned and expressed as recombinant proteins in E. coli. HgammaS and BgammaS are found to be authentic according to their spectral and hydrodynamic properties. Both full-length proteins and isolated domains are monomeric and exhibit high thermal and pH stabilities. The thermodynamic characterization made use of chemically and thermally-induced equilibrium unfolding transitions at varying pH. In spite of its exemplary two-domain structure, gammaS-crystallin does not show bimodal unfolding characteristics. In the case of BgammaS, at pH 7.0, the C-terminal domain is less stable than the N terminal one, whereas for HgammaS the opposite holds true. Differential scanning calorimetry confirms the results of chemically-induced equilibrium unfolding transitions. Over the whole pH range between 2.0 and 11.5, HgammaS-crystallin and its isolated domains (HgammaS-N and HgammaS-C) follow the two-state model. The two-state unfolding of the intact two-domain protein points to the close similarity of the stabilities of the constituent domains. Obviously, interactions between the domains do not contribute significantly to the overall stability of gammaS-crystallin. In contrast, the structurally closely related gammaB crystallin owes much of its extreme stability to domain interactions. PMID- 11026676 TI - Structure-function studies of the non-heme iron active site of isopenicillin N synthase: some implications for catalysis. AB - Isopenicillin N synthase (IPNS) is a non-heme ferrous iron-dependent oxygenase that catalyzes the ring closure of delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D valine (ACV) to form isopenicillin N. Spectroscopic studies and the crystal structure of IPNS show that the iron atom in the active species is coordinated to two histidine and one aspartic acid residues, and to ACV, dioxygen and H2O. We previously showed by site-directed mutagenesis that residues His212, Asp214 and His268 in the IPNS of Streptomyces jumonjinensis are essential for activity and correspond to the iron ligands identified by crystallography. To evaluate the importance of the nature of the protein ligands for activity, His214 and His268 were exchanged with asparagine, aspartic acid and glutamine, and Asp214 replaced with glutamic acid, histidine and cysteine, each of which has the potential to bind iron. Only the Asp214Glu mutant retained activity, approximately 1% that of the wild type. To determine the importance of the spatial arrangement of the protein ligands for activity, His212 and His268 were separately exchanged with Asp214; both mutant enzymes were completely defective. These findings establish that IPNS activity depends critically on the presence of two histidine and one carboxylate ligands in a unique spatial arrangement within the active site. Molecular modeling studies of the active site employing the S. jumonjinensis IPNS crystal structure support this view. Measurements of iron binding by the wild type and the Asp214Glu, Asp214His and Asp214Cys-modified proteins suggest that Asp214 may have a role in catalysis as well as in iron coordination. PMID- 11026677 TI - A universal thermodynamic approach to analyze biomolecular binding experiments. AB - Binding processes of any kind can be characterized as an association of a given ligand with some binding factor. This includes macromolecules as well as supramolecular aggregates such as micelles or membranes. The underlying molecular binding mechanism may be more or less complicated due to various intermediate steps (involving for instance conformational changes, aggregation, cooperativity, etc.). A sensible discussion of possible binding models naturally calls for a model-independent access to basic thermodynamic properties. The present contribution will demonstrate how this can quite generally be accomplished by a pertinent processing of properly selected experimental data. The method requires a series of titration measurements comprising the use of variable amounts of both the ligand and the binding factor. It leads to a linear mass conservation plot (i.e. amount of the ligand vs. a matching amount of the binding factor) whose slope and ordinate intercept are equal to the binding ratio (i.e. bound ligand per binding factor) and the free ligand concentration, respectively. This establishes the specific binding isotherm. The approach also reveals latent structurally determined features of the applied physical measuring signal. A number of examples including specific binding, unspecific adsorption and insertion in two-dimensional molecular films will illustrate the methodology. PMID- 11026678 TI - A universal system for the transport of redox proteins: early roots and latest developments. AB - The transport of proteins binding redox cofactors across a biological membrane is complicated by the fact that insertion of the redox cofactor is often a cytoplasmic process. These cytoplasmically assembled redox proteins must thus be transported in partially or completely folded form. The need for a special transport system for redox proteins was first recognized for periplasmic hydrogenases in gram-negative bacteria. These enzymes, which catalyze the reaction H2 <--> 2H+ + 2e, are composed of a large and a small subunit. Only the small subunit has an unusually long signal sequence of 30-50 amino acid residues, characterized by a conserved motif (S/T)-R-R-x-F-L-K at the N-terminus. This sequence directs export of the large and small subunit complex to the periplasm. Sequencing of microbial genes and genomes has shown that signal sequences with this conserved motif, now referred to as twin-arginine leaders, occur ubiquitously and export different classes of redox proteins, containing iron sulfur clusters, molybdopterin cofactors, polynuclear copper sites or flavin adenine dinucleotide. Mutations in an Escherichia coli operon referred to as mtt (membrane targeting and translocation) or tat (twin arginine translocation) are pleiotropic, i.e. these prevent the expression of a variety of periplasmic oxido reductases in functional form. The Mtt or Tat pathway is distinct from the well known Sec pathway and occurs ubiquitously in prokaryotes. The fact that its component proteins share sequence homology with proteins of the delta pH pathway for protein transport associated with chloroplast thylakoid assembly, illustrates the universal nature of this novel protein translocation system. PMID- 11026679 TI - Analytical ultracentrifugation and the characterization of chromatin structure. AB - This mini review consists of two parts. The first part will provide a brief overview of the theoretical aspects involved in the two kinds of experiments that can be conducted with the analytical ultracentrifuge (sedimentation velocity and sedimentation equilibrium) as they pertain to the study of chromatin. In the following sections, I describe the analytical ultracentrifuge experiments which, in my opinion, have contributed the most to our understanding of chromatin. Few other biophysical techniques, with the exception of X-ray scattering and diffraction, have contributed as extensively as the analytical ultracentrifuge to the characterization of so many different aspects of chromatin structure. In the course of his scientific career, Professor Henryk Eisenberg has made many important contributions to the theoretical aspects underlying ultracentrifuge analysis, especially in the analysis of solutions of polyelectrolytes and biological macromolecules [H. Eisenberg, Biological macromolecules and polyelectrolytes in solution, Clarendon Press, Oxford, 1976]. As an example he has devoted some of his research effort to the characterization of chromatin in solution. This review includes these important contributions. PMID- 11026680 TI - Halophilic enzymes: proteins with a grain of salt. AB - Halophilic enzymes, while performing identical enzymatic functions as their non halophilic counterparts, have been shown to exhibit substantially different properties, among them the requirement for high salt concentrations, in the 1-4 M range, for activity and stability, and a high excess of acidic over basic amino residues. The following communication reviews the functional and structural properties of two proteins isolated from the extremely halophilic archaeon Haloarcula marismortui: the enzyme malate-dehydrogenase (hMDH) and the 2Fe-2S protein ferredoxin. It is argued that the high negative surface charge of halophilic proteins makes them more soluble and renders them more flexible at high salt concentrations, conditions under which non-halophilic proteins tend to aggregate and become rigid. This high surface charge is neutralized mainly by tightly bound water dipoles. The requirement of high salt concentration for the stabilization of halophilic enzymes, on the other hand, is due to a low affinity binding of the salt to specific sites on the surface of the folded polypeptide, thus stabilizing the active conformation of the protein. PMID- 11026681 TI - Allostery in very large molecular assemblies. AB - In contrast to small allosteric systems (like hemoglobin) those containing very large numbers (n) of binding sites never exhibit cooperativity (as measured by the Hill coefficient, nH) even approaching the potential limit, n. The reason for this appears to be that in such macromolecules the cooperative unit always represents some sub-structure of the entire structure. On the other hand, it is frequently observed that such sub-structures, when isolated, do not exhibit cooperativity at all. This paper describes studies of some molluscan hemocyanins that explore this apparent anomaly. It is concluded that it is the higher order structure of the molecule that provides a framework within which the sub structures may exhibit their allosteric behavior. PMID- 11026682 TI - Structural and thermodynamic aspects of cooperativity in the homodimeric hemoglobin from Scapharca inaequivalvis. AB - The homodimeric cooperative hemoglobin from the mollusk Scapharca inaequivalvis displays an unusual subunit assembly with respect to vertebrate hemoglobins. The intersubunit contact region is formed by the two heme-carrying E and F helices, which bring the two hemes in contact with each other. At variance with tetrameric vertebrate hemoglobins, the ligand binding is not accompanied by a significant quaternary transition. The major ligand-linked changes are tertiary and are limited to the heme pocket and subunit interface. These unique structural features of HbI are not easily reconciled with the classical thermodynamic models used to describe cooperative ligand binding in vertebrate hemoglobins. The lack of distinct quaternary states and the absence of allosteric effectors suggested that cooperativity in HbI is entirely homotropic in origin. Thereafter, high resolution X-ray crystallographic data displayed the preferential binding of water molecules at the intersubunit interface in the unliganded protein with respect to the liganded one. These ordered water molecules were thus proposed to act as heterotropic effectors in HbI. The contribution of specific water binding to the observed cooperativity in HbI is discussed in the framework of the enthalpy-entropy compensation effect emerging from previous accurate equilibrium oxygen binding measurements. PMID- 11026683 TI - Extra-helical guanine interactions in DNA. AB - We review the extra-helical guanine interactions present in many oligonucleotide crystals. Very often terminal guanines interact with other guanines in the minor groove of neighboring oligonucleotides through N2 x N3 hydrogen bonds. In other cases the interaction occurs with the help of Ni2+ ions. Guanine/netropsin stacking in the minor groove has also been found. From these studies we conclude that guanine may have multiple extra-helical interactions. In particular it may be considered a very effective minor groove binder, which could be used in the design of sequence selective binding drugs. Interactions through the major groove are seldom encountered, but might be present when DNA is stretched. Such interactions are also analyzed, since they might be important for homologous chromosome pairing during meiosis. PMID- 11026684 TI - Intermolecular dynamics and function in actin filaments. AB - Structural models of F-actin suggest that three segments in actin, the DNase I binding loop (residues 38-52), the hydrophobic plug (residues 262-274) and the C terminus, contribute to the formation of an intermolecular interface between three monomers in F-actin. To test these predictions and also to assess the dynamic properties of intermolecular contacts in F-actin, Cys-374 pyrene-labeled skeletal alpha-actin and pyrene-labeled yeast actin mutants, with Gln-41 or Ser 265 replaced with cysteine, were used in fluorescence experiments. Large differences in Cys-374 pyrene fluorescence among copolymers of subtilisin-cleaved (between Met-47 and Gly-48) and uncleaved alpha-actin showed both intra- and intermolecular interactions between the C-terminus and loop 38-52 in F-actin. Excimer band formation due to intermolecular stacking of pyrene probes attached to Cys-41 and Cys-265, and Cys-41 and Cys-374, in mutant yeast F-actin confirmed the proximity of these residues on the paired sites (to within 18 A) in accordance with the models of F-actin structure. The dynamic properties of the intermolecular interface in F-actin formed by loop 38-52, plug 262-274 and the C terminus may account for the observed cross-linking of these sites with reagents < 18 A. The functional importance of actin filament dynamics was demonstrated by the inhibition of the in vitro motility in the Gln-41-Cys-374 cross-linked actin filaments. PMID- 11026686 TI - Structural dynamics of myoglobin. AB - Conformational fluctuations have been invoked to explain the observation that the diffusion of small ligands through a protein is a global phenomenon, as suggested (for example) by the oxygen induced fluorescence quenching of buried tryptophans. In enzymes processing large substrates, a channel to the catalytic site is often seen in the crystal structure; on the other hand in small globular proteins, it is not known if the cavities identified in the interior space are important in controlling their function by defining specific pathways in the diffusion to the active site. This point is addressed in this paper, which reports some relevant results obtained on myoglobin, the hydrogen atom of molecular biology. Protein conformational relaxations have been extensively investigated with myoglobin because the photosensivity of the adduct with CO, O2 and NO allows us to follow events related to the migration of the ligand through the matrix. Results obtained by laser photolysis, molecular dynamics simulations, X-ray diffraction of intermediate states of wt type and mutant myoglobins are briefly summarized. Crystallographic data on the photochemical intermediate of a new triple mutant of sperm whale myoglobin (Mb-YQR) show, for the first time, the photolyzed CO* sitting in one of the Xe-binding cavities, removed from the heme group. These results support the viewpoint that pre-existing 'packing defects' in the protein interior play a major role in controlling the dynamics of ligand binding, including oxygen, and thereby acquire a survival value. PMID- 11026685 TI - Electroporative fast pore-flickering of the annexin V-lipid surface complex, a novel gating concept for ion transport. AB - In contact with lipid bilayers and Ca2+-ions, the intracellular protein human annexin V (wild-type), Mr = 35,800, forms two types of cation-selective channels for the transport of Ca2+-, K+-, Na+- and Mg2+-ions, depending on the protein concentration [AN]. Type (I) channel events are large and predominant at high values [AN] > or = K = 5 nM at 296 K. At 50 mM Ca2+, symmetrical on both membrane sides, AN added at the cis side, the conductance is gCa(I) = 22 +/- 2 pS and at symmetrical 0.1 M K+-conditions: gK(I) = 32 +/- 3 pS, associated with two mean open-times tau1(I) = 0.68 +/- 0.2 ms and tau2(I) = 31 +/- 2 ms. Monoclonal anti AN antibodies added to the trans-side first increase the mean open-times and then abolish the channel activity, suggesting that type (I) channels refer to a membrane spanning protein complex, probably a trimer T, which at [AN] > K changes its membrane organization to a higher oligomer, probably to the side-by-side double-trimer T2. The smaller type (II) channel events are predominant at low [AN] < or = K and refer to the (electroporative) adsorption complex of the monomer. The conductances g(i)(II) for symmetrical concentrations depend non linearly on the voltage Um = Uext + U(AN), where U(AN) = 0.02 +/- 0.002 V is the electrostatic contribution of the Ca2+-AN complex and Uext the externally applied voltage. There is only one mean open-time tau(o)(II) which is voltage-dependent according to a functional of b x Um2 where b = 113.9 +/- 15 V(-2), yielding an activation Gibbs free energy of Ga = RT x b x Um2. The conformational flicker probability f(i)(II) in g(i)(II) = g(i)0(II) x gamma(i) x f(i)(II) is non linearly voltage-dependent according to a functional of a x Um2. The Nernst term gamma(i) refers to asymmetrical ion concentrations. From a = 50 V(-2), independent of the ion type, we obtain f(i)0(II) = 0.03 +/- 0.002 and the conductances for the fully open-channel states: gCa0(II) = 69 +/- 3 pS (0.05 M Ca2+) and gK0(II) = 131 +/- 5 pS (1.2 M K+). From the electroporation term a = pi[r(p)2]epsilon0(epsilon(w) - epsilon(m))/(2 kTd) we determine the mean pore radius of the complex in its fully open state as r(p)= 0.86 +/- 0.05 nm. The adsorbed annexin V (Ca2+) monomer appears to electrostatically facilitate the electric pore formation at the contact interface between the protein and the lipid phase. The complex rapidly flickers and thus limits the ion transport in a voltage-dependent manner. PMID- 11026687 TI - Transcription through nucleosomes. AB - Transcriptionally active genes in eukaryotes still retain most of the Chromatin packaging that is characteristic of eukaryotic DNA. Nucleosomes and even some higher order structure are present, although the histones may be chemically modified, for example by acetylation or phosphorylation, as part of the activation process. The presence of nucleosomes on the coding region of active genes raises the question: How does an RNA polymerase transcribe such a template? We have attempted to answer this question with relatively simple model systems involving a template carrying a single positioned nucleosome. We have shown that when a phage polymerase, SP6, transcribes such a template, the histone octamer of the nucleosome is not released into solution. Instead it is retained on the same DNA molecule, but displaced from its original binding site. Further studies have allowed us to propose a detailed model, which appears to hold not only for SP6 but also for transcription by the much larger RNA polymerase III from yeast. Our most recent results, obtained by electron cryomicroscopy, confirm and refine this model. PMID- 11026688 TI - Effects of excluded surface area and adsorbate clustering on surface adsorption of proteins I. Equilibrium models. AB - Statistical-thermodynamic models for the equilibrium adsorption of proteins onto homogeneous, locally planar surfaces are presented. An extension of earlier work [R.C. Chatelier, A.P. Minton, Biophys. J. 71 (1996) 2367], the models presented here allow for the formation of a broadly heterogeneous population of adsorbate clusters in addition to excluded volume interactions between all adsorbate species. Calculations are carried out for three simple models for the structure of adsorbate, illustrating similarities and differences in the equilibrium properties of maximally compact clusters, minimally compact clusters and isomerizing clusters. Depending upon the strength of attractive interactions between adsorbate molecules, the resulting equilibrium isotherms may exhibit negative cooperativity, positive cooperativity, essentially no apparent cooperativity, or a mixture of positive cooperativity at low surface density and negative cooperativity at high surface density of adsorbate. The condition of apparent lack of cooperativity, which might naively be interpreted as evidence of a lack of interaction between adsorbate molecules, actually conceals a balance between attractive and repulsive interactions and extensive clustering of adsorbate. PMID- 11026689 TI - Moist and soft, dry and stiff: a review of neutron experiments on hydration dynamics-activity relations in the purple membrane of Halobacterium salinarum. AB - Twenty-five years of neutron experiments on hydration and thermal dynamics in purple membranes of Halobacterium salinarum are reviewed. Neutron diffraction, elastic and quasielastic scattering, allowed to map the distribution of water and lipids and to measure thermal fluctuations and correlation times in the membranes, under various conditions of temperature, hydration and lipid environment. Strong correlations were established between dynamics parameters and the activity of bacteriorhodopsin (the purple membrane protein), as a light driven proton pump supporting the hypothesis that the influence of hydration on activity is in fact due to its effects on membrane thermal dynamics. Hydrogen deuterium labelling experiments highlighted stiffer and softer parts in the bacteriorhodopsin structure. The soft parts would allow the conformational changes involved in activity, while the stiffer ones may control a valve-like function in vectorial proton transfer. PMID- 11026690 TI - The unfolding of our understanding of RNA structure: a personal reflection. AB - In this article, I review how our research on RNA began, how it led us to demonstrate the single-stranded nature of RNA, and the ways in which it differs from double-stranded DNA. It was based on the development of a method for the isolation of undegraded rRNA and the observation that in rRNA preparations due to their viscosity behavior resemble a flexible, contractile coil. In support of this assumption, birefringence of flow measurements showed that rRNA solutions gave moderate positive values, which disappeared upon addition of salt. This is in contrast with DNA solutions where considerable negative birefringence persists even in the presence of salt. Further studies on RNA showed a close correlation of the ionic strength dependencies of optical rotation, optical density and hydrodynamic properties. These early results indicated that rRNA and tRNA possess a significant secondary structure. I then review the basis of the hairpin model for the secondary structure of RNA and finally, summarize current understanding of the tertiary structure of RNA. PMID- 11026691 TI - Extensive intra-tumor heterogeneity in primary human glial tumors as a result of locus non-specific genomic alterations. AB - Genomic changes are a hallmark of the neoplastic process. These range from alterations at specific loci and defined karyotypic changes which influence tumor behavior to generalized alterations exemplified by microsatellite instability. Generalized genomic changes within a tumor would be evidence in favor of the mutator hypothesis which postulates a role for such extensive changes during tumorigenesis. In this report, we have used the DNA fingerprinting technique of randomly amplified polymorphic DNA (RAPD) analysis to study genomic alterations within primary human astrocytic tumors (gliomas) in a locus non-specific manner. The RAPD fingerprinting profile of consecutive segments of tumors 2 mm across was studied; 17 astrocytic (high- and low-grade) tumors were sectioned end to end. Tissue from 50 consecutive sections, 40 microm thick (total 2 mm across), was pooled and taken to be a tumor compartment. DNA was subjected to RAPD amplification by 15 random 10-mer primers. A tumor segment was taken to have a DNA fingerprinting pattern different from others in the same specimen when its RAPD profile differed from others by at least one band of one RAPD reaction. All but one of the tumors showed compartments with a unique genetic profile, indicating genomic instability leading to widespread intra-tumor genetic heterogeneity. Eight tumors were also studied for loss of heterozygosity (LOH) of the p53 and D17S379 loci in the different segments as examples of alteration of specific tumor influencing loci. Three showed LOH of p53, which was limited to only one compartment of each tumor. The extensive intra-tumor genetic instability detected in this study is suggestive of the overall high rate of change in the genomes of tumors including those of a lower grade. It is hypothesized that some of these altered clones, which manifest as zones of heterogeneity in a solid tumor, may accumulate changes at loci known to influence tumor behavior, and thus clinical outcome. PMID- 11026692 TI - The prognostic significance of vascular endothelial growth factor (VEGF C-1) immunoexpression in oligodendroglioma. An analysis of 91 cases. AB - Oligodendrogliomas continue to generate considerable difficulties in identifying prognostic factors, including single histopathological patterns. Among the latter, vascular productive changes have been intensively examined but the obtained results appear to be controversial. Numerous studies have revealed an indisputable importance of vascular endothelial growth factor (VEGF) immunoreactivity for malignant progression of astrocytomas but the data regarding oligodendrogliomas are heterogeneous. Ninety-one patients with oligodendroglioma were examined retrospectively for VECF immunoexpression. The results demonstrated significant preponderance of intracellular VEGF expression for WHO grade III tumors. Nevertheless, VEGF staining patterns correlated with both progression free and overall survival only in univariate, but not in multivarite analysis. Only WHO tumor grade was found to be an independent prognostic factor for oligodendroglioma outcome. Therefore, it seems unlikely that VEGF immunohistochemistry will be of value in assessing individual oligodendroglioma prognosis, especially for determining of both progression-free and overall survival from histologically low-grade tumors. PMID- 11026693 TI - Soluble Fas-ligand (sFasL) in human astrocytoma cyst fluid is cytotoxic to T cells: another potential means of immune evasion. AB - Gliomas of all grades have been shown to express FasL, an apoptosis-inducing protein. Because of the ability of FasL to be cleaved from cell surfaces by metalloproteinases, soluble FasL can be released by FasL bearing cells into surrounding tissues. In the present study, we demonstrate the presence of sFasL in the cyst fluids of astrocytomas. Additionally, a human T-cell line, Jurkat, exposed to astrocytoma cyst fluid resulted in significantly increased cytotoxicity as compared to controls, an effect blocked by FasL neutralizing antibodies. This suggests that sFasL, may be utilized as a means of escaping immune surveillance by these tumors. PMID- 11026694 TI - Tolerance of the normal canine brain to epithermal neutron irradiation in the presence of p-boronophenylalanine. AB - Twelve normal dogs underwent brain irradiation in a mixed-radiation, mainly epithermal neutron field at the Brookhaven Medical Research Reactor following intravenous infusion of 950 mg of 10B-enriched BPA/kg as its fructose complex. The 5 x 10 cm irradiation aperture was centered over the left hemisphere. For a subgroup of dogs reported previously, we now present more detailed analyses including dose-volume relationships, longer follow-ups, MRIs, and histopathological observations. Peak doses (delivered to 1 cm3 of brain at the depth of maximum thermal neutron flux) ranged from 7.6 Gy (photon-equivalent dose: 11.8 Gy-Eq) to 11.6 Gy (17.5 Gy-Eq). The average dose to the brain ranged from 3.0 Gy (4.5 Gy-Eq) to 8.1 Gy (11.9 Gy-Eq) and to the left hemisphere, 6.6 Gy (10.1 Gy-Eq) to 10.0 Gy (15.0 Gy-Eq). Maximum tolerated 'threshold' doses were 6.7 Gy (9.8 Gy-Eq) to the whole brain and 8.2 Gy (12.3 Gy-Eq) to one hemisphere. The threshold peak brain dose was 9.5 Gy (14.3 Gy-Eq). At doses below threshold, some dogs developed subclinical MRI changes. Above threshold, all dogs developed dose-dependent MRI changes, neurological deficits, and focal brain necrosis. PMID- 11026695 TI - Cerebral vasculopathy and multiple infarctions in a woman with carcinomatous meningitis while on treatment with intrathecal methotrexate. AB - We report on a 33-year-old woman with carcinomatous meningitis due to carcinoma of the breast who developed multiple cerebral infarctions within four days after intrathecal chemotherapy with methotrexate. MR angiography revealed a narrowing of basal cerebral arteries, which is consistent with vasculopathy. The vasculopathy was probably due to carcinomatous meningitis itself, an acute toxic effect of methotrexate, or a combination of both. PMID- 11026696 TI - Primary leptomeningeal lymphoma with long-term survival: a case report. AB - Primary leptomeningeal lymphoma (PLML) is a rare disease. The most common presentation is symptoms of increased intracranial pressure. Confusion, dysarthria, hearing loss, paraparesis and lumbosacral spinal root symptoms have also been reported. Chemotherapy and radiotherapy have been tried, but its prognosis is usually poor. We experienced a case of PLML with a relatively benign course in an 18-year-old girl. Initial diagnosis was made as idiopathic intracranial hypertension. Lumbosacral shunt was done with good response for 3 years. When headache recurred, she was reevaluated and was correctly diagnosed as PLML. PMID- 11026697 TI - Efficacy of conventional radiotherapy for recurrent meningioma. AB - Results of radiation therapy for 20 patients with recurrent meningioma were analyzed. The patients included 8 men and 12 women, with a median age of 55 years. All of the patients had undergone at least one operation prior to the reoperation preceding radiotherapy. Ten patients had benign meningiomas, while 4 and 6 patients had atypical and malignant meningiomas, respectively, at the time of radiotherapy. The median radiation dose was 59.4 Gy (range: 50-61.2 Gy). The local control rate at 5 years was 36% for all 20 patients (41% for benign meningiomas and 30% for atypical or malignant meningiomas). The 5-year survival rate was 47%. Excluding 2 patients whose follow-up period was shorter than the preradiotherapy interval from the previous operation, the postradiation recurrence-free period was longer than the preradiotherapy interval in 50% (9/18) of the patients. No serious complications of radiotherapy were observed. Radiotherapy seemed to be effective in controlling the tumor or delaying recurrence in at least half of the patients. However, higher doses of radiation, using sophisticated radiation techniques, may be necessary to obtain higher control rates. PMID- 11026698 TI - Multiple metastases of carcinoma basocellulare into spinal column. AB - Basal cell carcinoma presents a relatively low potential and local malignancy and very slow growth giving only occasionally metastatic spreading. The frequency of occurrence of metastatic dissemination is estimated in the literature depending on examined population from 0.028% to 0.55%. Metastases are most often found in lymph nodes, lungs bones and internal organs: liver, spleen, kidneys, adrenal glands, pleura and the peritoneum. Authors present a case of a 69-years old female with an extensive basal cell carcinoma of the head convexity, infiltrating the subcutaneous tissue, periostium, bone and dura mater, giving distant metastases to other bone and soft tissue structures of a thoracic spine, which was confirmed by biopsy and histopathological findings of neoplasm tissue in spine. The primary lesion was successfully treated surgically. Despite administered radiotherapy of metastases in spine, progress of the disease during 1-year period was observed. The patient was alive with metastatic tumours present at last follow-up. Basing on the review of the literature and our case report we can distinguish following factors which may increase the risk of occurrence of basal cell carcinoma metastases: the great extent of the primary lesion, deep penetration to stromal tissue, blood and lymph vessel invasion, long history of tumour occurrence and the presence of metatypia in histopathological findings. The above-mentioned case fulfils the criteria of carcinoma basocellulare metastases proposed by Latters and Kessel and may be included to the general registration list of this cancer in the world. PMID- 11026699 TI - Accelerated radiotherapy with concomitant ACNU/Ara-C for the treatment of malignant glioma. AB - PURPOSE: To evaluate activity and toxicity of simultaneous ACNU and Ara-C with concurrent accelerated hyperfractionated radiotherapy in the treatment of high grade glioma. PATIENTS AND METHODS: Thirty patients aged 23-71 years (median 47.5), 16 patients with glioblastoma multiforme (GBM) and 14 patients with grade III glioma, received 93 courses of ACNU/Ara-C (median 4 courses) at following dose levels (ACNU/Ara-C in mg/m2/day): 70/90 (11 courses), 75/100 (36 courses) and 90/120 (46 courses). ACNU was administered IV on day 1 of each cycle, Ara-C as a 2 h-intravenous infusion on days 1-3. Patients received concomitant radiation therapy with 2 daily fractions of 1.75 Gy up to 57 Gy (median). RESULTS: Median survival of all patients was 13 months, 11 months for GBM and > 28 months for grade-III glioma; 31% (9 patients) survived longer than 24 months. The percentage of grade IV hematological toxicity was dose-dependent: 33% at the 70/90 dose level, 40% at 75/100 and 58% at 90/120. Six patients required platelet transfusion, 1 patient red blood cells; no febrile neutropenia occurred. Among 18 patients evaluable for response, 3 (17%) showed PR, 8 (44%) NC and 7 (39%) PD at completion of chemoradiation. No acute or late neurological toxicity occurred in this study. Younger age (p = 0.0001) and grade-III histology (p = 0.0009) were important prognostic factors for prolonged survival. CONCLUSION: This chemoradiation regimen is active in malignant gliomas and can be safely recommended at a dose level using 70 mg/m2 ACNU together with 90 mg/m2 Ara-C. PMID- 11026700 TI - Neurological dysfunction associated with postoperative cerebellar mutism. AB - BACKGROUND AND OBJECTIVES: The postoperative cerebellar mutism syndrome (CMS) is an unique acute postoperative complication characterized by transient decrease in speech output (often mutism), apathy, irritability as well as global cerebellar dysfunction. As much as 25% of patients undergoing a resection of a cerebellar or IV ventricular tumor may develop such a syndrome. In this retrospective study we characterize the clinical features of the CMS and explore potential etiologic mechanisms. METHODS: We conducted a retrospective analysis of medical records and imaging tests of 8 consecutive patients with the CMS identified through the database of the Children's Hospital and Dana-Farber Cancer Institute, Boston, and compared with a control group of 8 unaffected children undergoing a comparable tumor resection. RESULTS: In contrast to the control group, children in the affected group had marked decrease in speech output and comprehension, apathy and lack of initiative, inattention, persistent eye closure, flaccid hemiparesis and a severe global cerebellar dysfunction. Swallowing difficulties and bowel and bladder dysfunction were also observed. The median duration of the syndrome as judged by the persistence of the communication abnormalities was 4 weeks. The recovery was near complete with exception for a persistent global cerebellar dysfunction. A comparison of CT and MRI scans of children in both groups failed to identify distinguishing features. CONCLUSION: A surgical lesion of the midline cerebellum can cause a complex neurological dysfunction such as the CMS. Thus, we postulate that the cerebellum and its connections function as a 'modulatory system' in control of both motor and non-motor functions, including attention and language. PMID- 11026701 TI - The relationship between gastroesophageal reflux, intestinal metaplasia and adenocarcinoma of the esophagus. AB - Currently available data indicate a clear and probably causal relationship between long-lasting gastroesophageal reflux disease, the development of long segments with specialized intestinal metaplasia in the distal esophagus and subsequent progression to adenocarcinoma. To a lesser degree, this also appears to be the case for short segments of specialized intestinal metaplasia in the distal esophagus. In contrast, epidemiological data and classic parameters for the diagnosis of gastroesophageal reflux disease do not currently support a causal role of gastroesophageal reflux in the pathogenesis of specialized intestinal metaplasia at the gastric cardia. Despite its high prevalence and malignant potential, many questions about the prevention and management of intestinal metaplasia in the distal esophagus remain unsolved. In patients with chronic gastroesophageal reflux, current modes of medical therapy do not appear to prevent the development of intestinal metaplasia, while effective anti-reflux surgery seems to have a protective effect. Formal studies with adequate follow-up are, however, still lacking. Neither acid-suppression therapy nor anti-reflux surgery, with or without mucosal ablation, can reliably prevent the malignant degeneration of established intestinal metaplasia of the esophagus. Close endoscopic surveillance with extensive biopsies, therefore, remains mandatory in such patients, irrespective of the treatment modality. PMID- 11026702 TI - Results of short-and long-term medical treatment of gastroesophageal reflux disease (GERD). AB - INTRODUCTION: Symptomatic esophageal reflux affects a large number of individuals. Many find relief by avoiding trigger substances such as coffee or sweets; in other cases, lifestyle modifications do not suffice and drug treatment is necessary for symptom control. An adequate classification of gastroesophageal reflux disease (GERD) is currently lacking; esophagitis can be graded according to Savary and Miller or the more recent metaplasia ulcer stricture erosions (MUSE) classification. TREATMENT AIMS: The control of symptoms should be achieved in all patients: in addition, if esophagitis is present, the healing of erosions/ulcers as well as the prevention of further complications, such as strictures, hemorrhage, Barrett's esophagus or ulceration, must be accomplished. SHORT-TERM TREATMENT: In the case of rare symptoms, control might be achieved by lifestyle modifications and by antacids or mucosal protectants taken on demand. In the case of continuous symptoms or signs of esophagitis, effective inhibition of gastric acid secretion with proton pump inhibitors (PPIs) is necessary in many patients. PREVENTION OF RELAPSE: After discontinuation of medical therapy, almost all patients with esophagitis will experience a relapse within 30 weeks. The regimen offering the highest rate of remission in these patients is the one that induced remission in the first place. Reduction of PPI dose or a switch to H2 receptor antagonists increases the rate of relapse. RISKS OF LONG-TERM TREATMENT: Long-term acid suppressive therapy, as with the use of PPIs, may lead to hypergastrinemia, a situation in which the endocrine cells of the stomach may proliferate. In the presence of Helicobacter pylori infection, PPIs are more efficient in healing esophagitis; however, the occurrence of gastric mucosal atrophy, a potentially pre-cancerous condition, has been described. To date, however, no case of gastric cancer or endocrine neoplasia associated with PPI treatment has been documented; gastric mucosal atrophy is more likely to result from H. pylori infection and gastric carcinoid formation needs a genetic predisposition, such as multiple endocrine neoplasia (MEN) type I. CONCLUSIONS: Most cases of GERD can be effectively treated by non-surgical measures; in patients presenting with warning symptoms or persistent heartburn, endoscopy of the upper gastrointestinal tract is mandatory. Long-term use of PPIs seems to be a safe and efficient treatment for GERD. For the prevention of relapse, similar doses are needed as for the induction of remission in reflux esophagitis. PMID- 11026703 TI - Laparoscopic fundoplication--short- and long-term outcome. AB - Gastroesophageal reflux disease is probably the most frequently occurring benign functional disorder in the Western industrial countries. With the increasing popularity of laparoscopic anti-reflux procedures, issues on the appropriate technique have been revitalized. The purpose of this study is to evaluate the short- and long-term outcomes of laparoscopic fundoplication and reflect on the perspective of an increasing frequency of performed operations. The data sampling is based on a literature review and a questionnaire. It can be summarized that reflux recurrence due to breakdown of the wrap or herniation of the wrap can also develop in later years after the primary surgery and amount up to 8%. Persistent dysphagia is a severe problem in the first post-operative year, but usually decreases with time and is limited to rates of 3-5% on the long-term follow-up. Other functional problems, such as gasbloat, meteorism and epigastric pain--the cause often cannot be further detected or specified--limit the quality of life of patients after laparoscopic anti-reflux surgery in the long-term follow-up in up to 5% of cases. Side effects of laparoscopic antireflux procedures can be limited to 5 to 10%, but not totally avoided. PMID- 11026704 TI - Therapeutic experience of 65 cases with organic hyperinsulinism. AB - INTRODUCTION: The treatment for insulinoma is enucleation or resection, which are associated with various degrees of morbidity, including fistulas, pseudocysts, and necessity for reoperations. PATIENTS: We evaluated the outcome of 65 patients operated on for organic hyperinsulinism at an experienced university hospital (Department of Surgery, Heinrich-Heine University, Dusseldorf, Germany) between 1990 and 1998. Procedures included enucleation (n=37), left-sided resection (n=19), combination of enucleation from the head and left-sided resection (n=4), subtotal left pancreatectomy (n=2), and pancreaticoduodenectomy (n=3). RESULTS: In 47 out of the 65 patients (72%), the expected benign solitary insulinoma was found. Seven patients had malignant tumors and another eight had multiple tumors (including two with multiple endocrine neoplasia type 1 (MEN-1)), and three had nesidioblastosis. In total, 20 patients (31%) developed postoperative drainage of high amylase containing fluid, which in all but three cases resolved with conservative therapy. The three patients who developed fistulas and the three patients who developed pseudocysts underwent reoperative surgery. Seventeen (41%) of the patients who underwent enucleation (10 from the pancreatic head; 50%) and six (25%) of the resected patients demonstrated these complications. Postoperative glucose metabolism was normalized in all patients. CONCLUSION: Resection, especially in the head of the pancreas, demonstrates lower complication rates than enucleation. We propose resection for tumors that are large, malignant, situated close to pancreatic duct, and are multiple, including in MEN-1. For tumors in the head, pancreaticoduodenectomy may be an occasional choice. PMID- 11026705 TI - Cytokeratin-positive cells in bone marrow in comparison with other prognostic factors in colon carcinoma. AB - BACKGROUND AND AIMS: Despite the use of radical locoregional therapeutic methods and although conventional methods of diagnosis give no indication of metastases at the time of operation, distant metastases develop in approximately 50% of carcinoma patients within 5 years. While local relapses after the R0 resection of solid tumors are mainly a matter of concern for the surgeon, distant metastases can be traced back to the systemic dissemination of tumor cells at the time of operation. PATIENTS/METHODS: A prospective study is presented in which 145 patients suffering from colon carcinoma were analyzed for the prognostic relevance of isolated disseminated tumor cells detected in the bone marrow (IDT BM). The patients were operated on between 1993 and 1997 and subsequently observed until 1999. RESULTS: The monoclonal antibody A45-B/B3 was used with the immunocytochemical standard method for detecting IDT BM. For the purpose of cell cultivation, the cells were marked with the HEA-125 antibody and separated by means of magnetic cell sorting (MACS). CONCLUSION: In this investigation the presence of isolated disseminated tumor cells, as indicated by the A45-B/B3 antibody, proved to be an independent prognostic factor for survival time. The risk of an earlier death increased in node-negative and metastases-free patients with the detection of IDT BM by a factor of 12.60. The detection of IDT BM also represented an independent prognostic factor for the time until advancement of the tumor. The risk of an earlier relapse increased with the detection of disseminated tumor cells in the bone marrow containing the A45-B/B3 antibody by a factor of 18.02. A generally acknowledged standardization of the method is desirable. Due to the importance of the independent prognostic IDT BM factor, this method of ascertaining the pathological stage should be established at institutions of higher learning. PMID- 11026706 TI - Perforated peptic ulcer: is there a difference between Eastern Europe and Germany? Copernicus Study Group and Acute Abdominal Pain Study Group. AB - BACKGROUND AND AIMS: Ulcer surgery and the epidemiology of peptic ulcer perforation have changed considerably in recent decades. PATIENTS/METHODS: Within two prospective studies, 170 perforated peptic ulcer patients from 12 Eastern European centres and 37 patients from 11 German centres were analysed. RESULTS: The median age of patients was 43 years in the Copernicus study and 49 years in the MEDWIS study (P=n.s.), being higher for MEDWIS female patients (73 vs 53 years, respectively; P<0.05). Female patients made up 17% (29/170) of the Copernicus study and 35% (40/170) of the MEDWIS study (P<0.05). Twenty-three per cent (40/170) of patients in the Copernicus study and 54% (20/37) in the MEDWIS study had gastric ulcer perforation (P<0.001). The proportion of definitive operations was higher in Eastern Europe (41.1%; 67/163) than it was in Germany (16.1%; 5/31) (P<0.01). German patients experienced more general complications than Eastern European patients (35 vs 12%, respectively; P<0.01) and a higher mortality [13% (5/37) vs 2% (4/170), respectively; P<0.01]. Delayed admission > or =12 h and age > or =60 years remained predictors for complications in multivariate logistic regression analysis. CONCLUSION: The proportion of both women and gastric ulcers was higher among German patients, while Eastern European patients underwent more definitive operations. German patients experienced more general complications and a higher mortality. Complications were related to high age and delayed admission. PMID- 11026707 TI - Hepato-venous reconstruction in orthotopic liver transplantation with preservation of the recipients' inferior vena cava and veno-venous bypass. AB - BACKGROUND AND AIMS: The potential advantages of vena cava-preserving recipient hepatectomy in orthotopic liver transplantation are reduced hemorrhage, improved cardiovascular stability and preserved renal perfusion without the requirement of veno-venous bypass as compared with recipient hepatectomy including the vena cava. No detailed information is available on the use of veno-venous bypass during complicated vena cava preserving recipient hepatectomy and liver transplantation. In the present study, the peri- and postoperative courses of adult liver transplant recipients in whom the hepatovenous reconstruction was performed according to three different techniques with and without the use of veno-venous bypass were investigated. PATIENTS/METHODS: During primary orthotopic liver transplantation, an end-to-end (ETE) cavo-caval interposition of the donor vena cava to the recipient's vena cava was performed in 75 patients (group I). In 15 patients, a termino-terminal piggyback (PB) anastomosis was constructed to the remnant of the recipient's hepatic vein (group II), and in 72 transplantations a latero-lateral cavo-cavostomy (LLC) of donor-to-recipient's vena cava (group III) was performed. The use of bypass, operative time and cold ischemia time, perioperative blood product requirements, incidence of relaparotomy, the evolution of postoperative renal function, technical complications and the survival were analyzed and compared using multivariate statistics and actuarial techniques for statistical evaluation. RESULTS: No differences could be found in preoperative patient conditions, donor conditions, operating time, anastomosing time or cold ischemia time. In groups I-III, the veno-venous bypass was used in 50 (67%), 8 (53%) and 6 (8%) cases respectively (P=0.02 for group III). The mean preoperative packed cells requirements were 20.4 vs 29.6 vs 10.8 units (P=0.01 for group III), while postoperative blood product requirements (first 24 h) were 2.6 vs 5.0 vs 0.20 units of packed cells (P=0.02 for group III). Relaparotomy for diffuse retroperitoneal hemorrhage was performed 14 times (19%) in group I, 3 times (20%) in group II and 7 times (8.3%) in group III (P=0.002). The incidence of posteropative early renal dysfunction (increase of > or =1.3 mg% serum creatinine) in group I vs group II vs group III was 24% vs 60% vs 16.7% (P=0.001 for group II) for patients without the use of veno-venous bypass. No significant difference was observed concerning early renal dysfunction in patients where a veno-venous bypass was used. The survival at 12 months was 81% for group I, 86% for group II and 93.0% for group III. In group III there were four complications (P=0.03) at the hepatovenous anastomosis of which two were eventually fatal. CONCLUSION: Preservation of the recipient's vena cava and LLC can reduce, but not avoid, the requirement for veno-venous bypass. In orthotopic liver transplantation, postoperative hemorrhage, as measured by surgical revisions and requirement for blood products, is significantly reduced with LLC with and without bypass. Early renal dysfunction also occurs in the group of LLC as compared with the termino-terminal cavostomy independent of the bypass. A technical failure resulting in patient death can be associated with LLC. PMID- 11026708 TI - Differential effects of neurotensin and cholecystokinin on intestinal microcirculation after ischemia-reperfusion. AB - BACKGROUND: We investigated the effect of neurotensin and cholecystokinin (CCK) on intestinal microcirculation after ischemia-reperfusion. METHOD: Ischemia was induced in Wistar rats by occlusion of the superior mesenteric artery for 40 min. Ten minutes before reperfusion, infusion of either neurotensin or CCK was started. Afterwards, the microhemodynamics of the jejunum were examined by means of intravital microscopy. RESULTS: Ischemia-reperfusion decreased functional capillary density from 873.4+/-18.1 to 362.5+/-8.3 cm(-1) and red blood cell velocity from 0.49+/-0.03 to 0.34+/-0.02 mm/s. Furthermore, leukocyte-endothelium interaction was increased. Neurotensin infusion significantly increased functional capillary density to 483.2+/-9.0 cm(-1) and red blood cell velocity to 0.69+/-0.01 mm/s in the mucosal capillaries compared with ischemic controls. Despite the amelioration of villus perfusion, the number of non-perfused villi significantly increased (11.8+/-3.6%) compared with ischemic controls. CCK infusion also resulted in a significant increase of functional capillary density (535.2+/-7.4 cm(-1)) and red blood cell velocity (0.67+/-0.01 mm/s). In contrast to neurotensin, the number of non-perfused villi was not increased (5.8+/-2.2%). CONCLUSION: We conclude that neurotensin further aggravates perfusion inhomogeneity and stasis when administered during the ischemic period. In contrast, CCK has no negative influence on perfusion homogeneity after ischemia reperfusion. It may be superior to neurotensin in the reconstitution of normal microvascular perfusion patterns after ischemia-reperfusion. PMID- 11026709 TI - Laparoscopic hepatic wedge resection of hemangioma: report of two cases. AB - INTRODUCTION: Cavernous hemangiomas are the most frequent type of benign liver tumor. A large proportion are discovered unexpectedly. The widespread use of ultrasound (US) and computerized tomography (CT) scanning has made diagnosis more common. Laparoscopic liver surgery has, however, developed more slowly. There have been only a few anecdotal reports of hepatic laparoscopic resections, most of which are limited to wedge resections. Laparoscopic anatomical liver resections are still at an early stage of development. DISCUSSION: This paper describes two cases of hepatic cavernous hemangiomas, both of which were removed laparoscopically. No blood transfusion was necessary. No surgical complications occurred and the patients were discharged on the second postoperative day. We conclude that, depending on the size and location of the tumor, laparoscopic resection of liver hemangiomas can be performed safely. PMID- 11026710 TI - MRI in inflammatory myopathies. AB - Inflammatory myopathies encompass a group of acquired muscle disorders caused by infectious agents (bacteria, viruses, fungi and parasitic agents) or autoimmune processes (polymyositis, dermatomyositis and other types). In suspected infection sonography, CT and MRI are all able to show edema and fluid collections in soft tissues and muscles; sonography and CT may help guidance of a needle aspiration to establish a correct diagnosis. By offering better tissue differentiation, MRI appears to be more efficient than sonography and CT in diagnosing and managing autoimmune myopathies. MRI is indeed very sensitive to the presence of water and edema, and appears to be a very good indicator for an early diagnosis of diseases. MRI may also help to evaluate the extent and number of lesions, to guide a biopsy in an area of active disease and finally to follow the evolution under therapy. PMID- 11026711 TI - MR imaging of septic sacroiliitis. AB - OBJECTIVE: To investigate the diagnostic value of magnetic resonance (MR) imaging in detecting septic sacroiliitis and to determine whether the MR characteristics allow this entity to be differentiated from sacroiliitis in spondylarthropathy (SpA). PATIENTS AND DESIGN: The imaging findings of 11 patients with septic sacroiliitis were retrospectively analyzed by two experienced radiologists. Radiographic surveys of the pelvis as well as computed tomography (CT) and MR images of the sacroiliac joints were available in all cases. Seven of the patients additionally underwent a follow-up MR examination. The MR imaging protocol comprised combinations of coronal and transverse T1-weighted spin-echo (SE) or fast SE sequences, T2-weighted gradient-echo (GE) sequences and short tau inversion recovery sequence (STIR) sequences as well as dynamic contrast-enhanced T1-weighted acquisitions. RESULTS: Three patients with a short disease history showed anterior and/or posterior subperiosteal infiltrations ("lava cleft phenomenon"), transcapsular infiltrations of juxtaarticular muscle layers, which obscured the fasciae, and periarticular bone marrow edema. The eight patients with more advanced stages of sacroiliitis additionally showed abscess formation, sequestration, and erosion. At follow-up MR examination (n=7) under systemic antibiotic treatment, the morphologic characteristics showed progression (n=1), regression (n=4), unchanged findings (n=1), or a mixed response (n=1). Clinical improvement precedes resolution of the MR findings. CONCLUSIONS: Anterior and/or posterior subperiosteal infiltrations and transcapsular infiltrations of juxtaarticular muscle layers were depicted in all patients. These MR imaging findings are characteristic of septic sacroiliitis and may be used to differentiate this entity from sacroiliitis in SpA. PMID- 11026712 TI - Tuberculosis of the sternum and clavicle: imaging findings in 15 patients. AB - OBJECTIVE: To describe the imaging findings in sterno-clavicular tubercular involvement. DESIGN AND PATIENTS: Fifteen patients with pathologically proven tuberculosis of the sternum and clavicle were retrospectively evaluated. Routine radiography, computed tomography (CT) and magnetic resonance imaging (MRI) were used in some or all of the patients. Clinical information and imaging features were evaluated in each case. RESULTS: Eight patients had sternoclavicular joint (SCJ) involvement, five had isolated sternal involvement and two had isolated clavicular involvement. Seven patients were evaluated with only CT, six with only MRI and two with both. There were eight male and seven female patients, varying in age between 16 and 78 years. Fever, swelling and pain were common presenting symptoms. Two patients were HIV positive. Radiographs were positive in only eight patients. Destruction and signal intensity (SI) changes of the sternum and clavicle, destruction of the cartilage, soft tissue changes representing granulation tissue/abscess, displacement of the adjacent structures (vessels, trachea, etc.) and inflammatory changes in the adjacent structures in the form of cellulitis and myositis were common imaging features. CONCLUSIONS: All imaging methods can provide complementary information regarding sterno-clavicular tubercular involvement that is helpful for determination of the therapy. MRI is useful in determining the extent of the lesion, particularly marrow involvement and soft tissue extent. PMID- 11026713 TI - Are L5 fractures an indicator of metastasis? AB - OBJECTIVE: To determine whether L5 vertebral body fractures are an indicator of malignancy. DESIGN AND PATIENTS: A retrospective study of L5 vertebral body fractures was carried out using plain radiographs, CT, and/or MRI. Over a 5-year period, 51 patients with L5 vertebral body fractures were seen at our institution. Since L1 vertebral body fractures are common, 51 age- and gender matched (20 men, 31 women; mean age 60 years) patients with L fractures were utilized as the control group. The frequency of neoplastic infiltration of the vertebrae was compared between these two populations to determine whether pathologic fracture was more frequent at L5. RESULTS: Twelve (24%) of the L5 fractures were pathologic compared with four (8%) of the L1 fractures (chi-square test, P<0.05). Neoplasm types included multiple myeloma (n=4), prostate (n=3), breast (n=2), lung (n=2), melanoma (n=2), bladder, colon, and leukemia (each n=1). CONCLUSION: Although most L5 fractures are not pathologic, there is an increased incidence of pathologic fractures in this location compared with L1. Therefore, a fracture of L5 should raise the suspicion of metastasis. PMID- 11026714 TI - Chondro-osseous differentiation in fat tissue tumors: magnetic resonance imaging with pathological correlation. AB - Chondro-osseous differentiation of three benign or malignant fat tissue tumors- two chondrolipomas and a liposarcoma with cartilaginous metaplasia--was studied with magnetic resonance (MR) imaging and compared with their pathological findings. The results suggest that demarcation of cartilage tissue can be clearly defined on MR imaging when the size of the cartilaginous area is large. Myxoid matrix, degenerative fat tissue and lipodystrophic change may decrease the delineation of the cartilage tissue. PMID- 11026715 TI - Juxtacortical chondromyxoid fibroma arising in an apophysis. AB - We present a rare case of juxtacortical chondromyxoid fibroma arising in the lesser trochanter of the right femur which corresponds to an apophysis. Radiography showed a well-defined expansive lesion with a sclerotic margin measuring 5x3.5 cm in diameter in the lesser trochanter. On spin echo T1-weighted images, the lesion revealed low signal intensity similar to muscle. On spin echo T2-weighted images, the lesion revealed high heterogeneous signal intensity, which after gadolinium injection showed heterogeneous enhancement. The inner margin of the cortex was intact and adjacent bone marrow was of normal signal intensity. The outer margin of the lesion was also clearly defined and extension into adjacent soft tissue beyond the exophytic cortical outgrowth was not evident. PMID- 11026716 TI - Solid aneurysmal bone cyst in the humerus. AB - We report on a 69-year-old woman with a solid variant of aneurysmal bone cyst (solid ABC) in the left humerus with a pathological fracture. Radiographically, the lesion exhibited a relatively well-defined osteolytic lesion in the diaphysis of the left humerus. On magnetic resonance (MR) imaging, the medullary lesion exhibited a homogeneous signal intensity isointense with surrounding normal muscles on the T1-weighted images and a mixture of low and high signal intensity on the T2-weighted images. Contrast-enhanced T1-weighted images revealed diffuse enhancement of the entire lesion. The pathological study showed a proliferation of fibroblasts, histiocytes, chronic inflammatory cells and numerous multinucleated giant cells in a collagenous matrix. Abundant osteoid formation in the matrix was observed, but the cells were devoid of nuclear atypia. Aneurysmal cystic cavities were absent. A review of the English literature found 22 cases of solid ABC of the long bones. PMID- 11026717 TI - Primary osteosarcoma of a metatarsal bone. AB - Although osteosarcoma is the most common primary bone malignancy of childhood and adolescence that is not related to marrow cells, involvement of the short tubular bones is uncommon. In contrast to more conventional sites, where the tumor is usually high grade and found in adolescents, osteosarcoma of the small bones is more likely to be low grade, and is often seen in older individuals. We present a case of low-grade primary osteosarcoma of a metatarsal bone in a 25-year-old woman. PMID- 11026718 TI - Diabetic muscle infarction: atypical MR appearance. AB - We describe a case of diabetic muscle infarction which had atypical features of hyperintensity of the affected muscle on T1-weighted images. Biopsy was performed which revealed diffuse extensive hemorrhage within the infarcted muscle. We believe increased signal intensity on T1-weighted images should suggest hemorrhage within the infarcted muscle. PMID- 11026719 TI - Multimodality imaging assessment of meniscal ossicle. AB - A case of meniscal ossicles occurring in the left knee of a 23-year-old woman is presented. Radiographs showed two calcified lesions at the posteromedial aspect of the knee which were interpreted as loose bodies. Sonography, computed tomography arthrography and magnetic resonance imaging showed the fragments within the posterior horn of the medial meniscus permitting a diagnosis of meniscal ossicles. These techniques can detect meniscal ossicles and exclude intra-articular loose bodies. PMID- 11026720 TI - Properties of utricular and saccular nerve-activated vestibulocerebellar neurons in cats. AB - Properties of otolith inputs to vestibulocerebellar neurons were investigated in 14 adult cats. In the vestibular nuclei, we recorded single-unit activities that responded orthodromically after stimulation of the utricular and/or saccular nerves and antidromically after stimulation of the cerebellum (uvula-nodulus and anterior vermis). Descending axonal projections to the spinal cord were also examined by antidromic stimulation of the caudal end of the C1 segment. Forty seven otolith-activated neurons that projected to the uvula-nodulus were recorded. Thirteen (28%) of the 47 neurons received convergent inputs from the utriculus and sacculus. The remaining 34 (72%) vestibular neurons were non convergent neurons: 18 (38%) received utricular input alone, and 16 (34%) received saccular input alone. Most (35/47) vestibulocerebellar neurons were located in the descending vestibular nucleus and only one of these projected to the spinal cord. Seven of the 47 vestibulocerebellar neurons were located in the lateral vestibular nucleus and most of these neurons projected to the spinal cord. The remaining neurons were located in group X (two neurons) and the superior vestibular nucleus (three neurons). In a different series of experiments, 37 otolith-activated vestibular neurons were tested to determine whether they projected to the uvula-nodulus and/or the anterior vermis. Nineteen of the 37 neurons projected to the anterior vermis, 13/37 projected to the uvula nodulus, and 5/37 projected to both. The utricular and/or saccular nerve activated vestibulocerebellar neurons projected to not only the uvulanodulus, but also to the anterior vermis. In summary, the results of this study showed that vestibular neurons receiving inputs from the utriculus and/or sacculus projected to the cerebellar cortex. This indirect otolith-cerebellar pathway terminated both in the anterior lobe and in the uvula/nodulus. PMID- 11026721 TI - Local and global stabilization of coordination by sensory information. AB - In studies of rhythmic coordination, where sensory information is often generated by an auditory stimulus, spatial and temporal variability are known to decrease at points in the movement cycle coincident with the stimulus, a phenomenon known as anchoring (Byblow et al. 1994). Here we hypothesize that the role of anchoring may be to globally stabilize coordination under conditions in which it would otherwise undergo a global coordinative change such as a phase transition. To test this hypothesis, anchoring was studied in a bimanual coordination paradigm in which either inphase or antiphase coordination was produced as auditory pacing stimuli (and hence movement frequency) were scaled over a wide range of frequencies. Two different anchoring conditions were used: a single-metronome condition, in which peak amplitude of right finger flexion coincided with the auditory stimulus; and a double-metronome condition, in which each finger reversal (flexion and extension) occurred simultaneously with the auditory stimuli. Anchored reversal points displayed lower spatial variation than unanchored reversal points, resulting in more symmetric phase plane trajectories in the double- than the single-metronome condition. The global coordination dynamics of the double-metronome condition was also more stable, with transitions from antiphase to inphase occurring less often and at higher movement frequencies than in the single-metronome condition. An extension of the Haken-Kelso-Bunz model of bimanual coordination is presented briefly which includes specific coupling of sensory information to movement through a process we call parametric stabilization. The parametric stabilization model provides a theoretical account of both local effects on the individual movement trajectories (anchoring) and global stabilization of observed coordination patterns, including the delay of phase transitions. PMID- 11026722 TI - Speed-accuracy trade-off in the performance of pointing movements in different directions in two-dimensional space. AB - Nine healthy subjects performed 2D pointing movements using a joystick that controlled a screen cursor. Continuous visual feedback was provided until movement completion. Three variables were systematically manipulated: (1) target distance, (2) target size and (3) target direction. A four-way factorial ANOVA was used to analyze the effects of these fixed factors and of the random factor of subject on several movement parameters. Movement time increased with increasing distance and decreasing target size and as predicted from Fitts' law. The target direction did not affect movement time. In contrast the direction, distance and size of the target significantly affected the movement time until the first zero crossing on the speed record reflecting the time to bring the arm into the vicinity of the target. Movements on the lateral axis of the horizontal plane (horizontal movements) resulted in a decrease in initial movement time compared to movements on the anterior axis of the horizontal plane (vertical movements). A significant effect of target distance and direction but not target size was observed for the magnitude of maximum acceleration, maximum speed and maximum deceleration. Horizontal movements had a larger maximum acceleration, speed and deceleration. Furthermore the maximum speed and deceleration occurred earlier in time for these horizontal movements. Finally the number of secondary peaks on the speed record increased with decreasing target size and was not affected by the target distance or target direction. In conclusion our results indicate that different movement parameters are affected by target distance, size and direction. The crucial distinction was between parameters affected by target size and direction. These parameters did not overlap. Target direction affects the first part of movement execution while target size affects the final part of movement execution. Thus a clear segmentation of movement execution in two phases is supported by these results. The implications of these results for theoretical models of speed-accuracy trade-off are discussed. PMID- 11026723 TI - GABA(B) receptors contribute to vestibular compensation after unilateral labyrinthectomy in pigmented rats. AB - The horizontal vestibulo-ocular reflex was studied in pigmented rats, which had been unilaterally, chemically labyrinthectomised 6-144 days previously. During this partially compensated stage after unilateral labyrinthectomy (UL), both static and dynamic deficits remain. The former was evaluated by recording of spontaneous eye movements in darkness, and the latter by estimating the slow phase velocity (SPV) gain of compensatory eye movements during horizontal vestibular stimulation. The GABA(B) agonist baclofen caused a reversal of the remaining ipsilesional drift of the eyes in darkness into a nystagmus with a contralesional slow phase. The GABA(B) antagonist CGP 36742 caused a decompensation by exaggerating the remaining ipsilesional eye drift. Further, baclofen equilibrated or reversed the asymmetry between ipsi- and contralesional SPV gains during horizontal sinusoidal rotations at 0.2 Hz and 0.8 Hz. This was achieved by an increase in the ipsilesional gain and a decrease in the contralesional gain. The phase lead during sinusoidal rotation (0.2 Hz) was larger following rotation to the lesioned side than to the intact side in UL rats. This asymmetry was reversed by baclofen. CGP 36742 inhibited the effects of baclofen, while the antagonist per se aggravated SPV gain and phase lead asymmetries in UL rats during vestibular stimulation. Per- and post-rotatory nystagmus induced by velocity step stimulation revealed an imperfect velocity storage function in UL animals, which was modulated by baclofen. An investigation of the baclofen effect on SPV gain asymmetry during different time intervals after chemical UL showed a completely developed effect on the 6th day. Bilateral flocculectomy did not alter the effects of baclofen on UL animals. It is concluded that physiological stimulation of GABA(B) receptors contributes to minimise the vestibulo-oculomotor asymmetry during the partially compensated period after UL. Administration of an agonist or an antagonist changes the asymmetry towards the ipsi- or contralesional side, possibly by altering the spontaneous neuronal activity in the bilateral medial vestibular nuclei. The results are compatible with a hypothesis, supported by in vitro slice experiments, that the efficacy of GABA(B) receptors is up-regulated on the ipsilesional side and down-regulated on the contralesional side. PMID- 11026724 TI - Inhibition of return is supramodal: a demonstration between all possible pairings of vision, touch, and audition. AB - Inhibition of return' (IOR) refers to the delayed detection often found for targets at the same location as a preceding event. We examined whether IOR reflects a truly supramodal phenomenon, in an experiment designed to avoid criticisms of previous crossmodal research. We presented a random sequence of visual, tactile, and auditory targets to either the left or right of central fixation, and tested for IOR between targets in all three modalities when presented successively to the same versus different side. Speeded detection for targets in all three modalities was indeed slower if the preceding target had been presented from the same position, regardless of the modality, of this preceding target. These results demonstrate for the first time that IOR is truly supramodal. PMID- 11026725 TI - High resolution DC-EEG mapping of the Bereitschaftspotential preceding simple or complex bimanual sequential finger movement. AB - The present set of experiments investigated the Bereitschaftspotential (BP) preceding voluntary bimanual sequential simple (task 1) and complex movements (task 2) in supplementary/cingulate and primary motor areas (SCMA, MIs) using 64 channel direct current electroencephalography analysis in 16 right-handed healthy subjects. The results showed that: (1) onset times of BPs preceding the two tasks were significantly earlier at Cz than at C3 and C4, (2) the complex task induced significantly larger amplitudes than the simple task over the SCMA 1.1 s before EMG onset (BPI period), over the SCMA and both MIs for the BP2 period, extending from the SCMA and MIs to all frontocentral, central, centroparietal, and frontal areas during the motor potential period, (3) task difference prior to 0.96 s mainly appeared in the SCMA rather than in either MI, (4) the BP had a significantly larger amplitude in the SCMA than in the MIs, the differences being asymmetric between the left and the right hemisphere motor areas, and (5) the sinks of BP current source density (CSD) preceding the two tasks were found in the frontocentral midline; and the regions and intensities of CSD maps were larger and stronger in task 2 than they were in task I at the same times of the epoch. The results suggested that: (1) the SCMA and MIs participate in bimanual sequential simple or complex movements, (2) the SCMA appears to not only serve as a trigger command for voluntary movement but also seems to design the different motor modes, (3) the amplitude, duration, onset time, CSD region, and intensity of BP all increase with the level of complexity of the movement, (4) the greater the complexity of the action, the earlier the preparation and the larger the extent of activated neuronal populations in the SCMA, (5) activation of the SCMA occurred prior to that of the MI, and (6) the activation suggests an asymmetry between left and right MIs in simultaneous bilateral finger movement, but this asymmetry seems to be less pronounced for complex movements. PMID- 11026726 TI - The cytotoxicity of chronic neuroinflammation upon basal forebrain cholinergic neurons of rats can be attenuated by glutamatergic antagonism or cyclooxygenase-2 inhibition. AB - The proinflammagen lipopolysaccharide (LPS) was infused chronically (37 days) into the basal forebrain of rats. The current study determined whether the chronic administration of either a non-competitive N-methyl-D-aspartate- (NMDA-) sensitive receptor antagonist, memantine, or a selective cyclooxygenase-2 (COX2)/lipoxygenase inhibitor, CI987, could provide significant neuroprotection from the cytotoxic effects of LPS-induced neuroinflammation. Chronic LPS infusions decreased cortical choline acetyltransferase activity, which paralleled a decline in the number of choline-acetyltransferase-immunoreactive-cells within the basal forebrain as well as the number of activated resident microglia. The infusions appeared to be selective for cholinergic neurons. Peripheral administration of memantine (i.p.) or CI987 (s.c.) significantly attenuated the cytotoxic effects of the chronic inflammatory processes upon cholinergic cells within the basal forebrain. However, only CI987 attenuated the neuroinflammation produced by LPS and the subsequent changes in microglial activation. These results indicate that the cytotoxic effects of chronic neuroinflammation may involve prostanoid synthesis and may operate through NMDA receptors, and that the effects of prostaglandins occur upstream to NMDA-receptor activation. PMID- 11026727 TI - Self-motion perception during a sequence of whole-body rotations in darkeness. AB - The main aim of this study was to examine how postrotatory effects, induced by passive whole-body rotations in darkness, could alter the perception of motion and eye movements during a subsequent rotation. Perception of angle magnitude was assessed in a reproduction task: blindfolded subjects were first submitted to a passive rotation about the earth-vertical axis on a mobile robot. They were then asked to reproduce this angle by controlling the robot with a joystick. Stimulus rotations ranged from 80 degrees to 340 degrees. Subjects were given one of two delay instructions: after the stimulus, they either had to await the end of postrotatory sensations before starting reproduction (condition free delay, FD), or they had to start immediately after the end of the stimulus rotation (no delay, ND). The delay in FD was used as an incidental measure of the subjective duration of these sensations. Eye movements were recorded with an infrared measuring system (IRIS). Results showed that in both conditions subjects accurately reproduced rotation angles, though they did not reproduce the stimulus dynamics. Peak velocities reached in ND were higher than in FD. This difference suggests that postrotatory effects induced a bias in the perception of angular velocity in the ND condition. PMID- 11026729 TI - Vision of the hand and environmental context in human prehension. AB - Previous findings on the role of visual contact with the hand in the control of reaching and grasping have been contradictory. Some studies have shown that such contact is largely irrelevant, while more recent ones have emphasised its importance. In contrast, information arising from the surrounding environment has received relatively little attention in the study of prehensile actions. In order to identify the roles of both sources of information, we made kinematic comparisons between three conditions. In the first, reaching was performed in a dimly lit room and compared with a second condition in which reaches in the dark, but with the thumb and first finger illuminated, were made to a luminous object. This contrast allows the effects of environmental context to be identified. A comparison between the second and a third condition, in which both vision of the hand and the environment was removed, but the object was still visually available, enabled the assessment of how and when vision of the hand plays a role. Removing environmental cues had effects both early and late in the reach, while vision of the hand was only crucial in the period after peak deceleration. In addition, removal of both sources of information resulted in larger grip apertures. Differences and similarities between our findings and those of other studies are discussed, as is the ongoing debate about the relative importance of visual feedback of the hand in the control and co-ordination of prehensile actions. We conclude with suggestions for further research based on the set-up used in the present study. PMID- 11026728 TI - Effects of intra-vestibular nucleus injection of the group I metabotropic glutamate receptor antagonist AIDA on vestibular compensation in guinea pigs. AB - Removal of the peripheral vestibular receptor cells in one inner ear (unilateral vestibular deafferentation, UVD) results in a syndrome of ocular motor and postural disorders, many of which disappear over time in a process of behavioural recovery known as vestibular compensation. Excitatory amino acid receptors, in particular the N-methyl-D-aspartate (NMDA) receptor, have been implicated in vestibular compensation; however, the metabotropic glutamate receptors (mGluRs) have not been studied in this context. The aim of this study was to determine whether group I mGluRs in the brainstem vestibular nucleus complex (VNC) ipsilateral to the UVD are involved in vestibular compensation of the static symptoms of UVD in guinea pig. The selective group I mGluR antagonist (RS)-1 aminoindan-1,5,dicarboxylic acid (AIDA) was continuously infused into the ipsilateral VNC for 30-min pre-UVD and 30-min post-UVD by cannula, at a rate of 1 microl/h, using one of four doses: 0.1 fg, 0.1 pg, 0.1 ng or 0.1 microg (n=5 animals in each case). In control conditions, a 0.1-fg (n=4) or 0.1-microg (n=5) NaOH vehicle was infused into the ipsilateral VNC using the same protocol. In order to control for the possibility that AIDA disrupted spontaneous neuronal activity in the VNC in normal animals, 0.1 microg AIDA (n=4) or 0.1 microg NaOH (n=2) was infused into the VNC in labyrinthine-intact animals. In both groups, static symptoms of UVD (i.e. spontaneous nystagmus, SN, yaw head tilt, YHT and roll head tilt, RHT) were measured at 8, 10, 12, 15, 20, 25, 30, 35, 45 and 50 h post-UVD. In addition, the righting reflex latency (RRL) was measured in labyrinthine-intact animals in order to assess whether AIDA impaired motor coordination in labyrinthine-intact animals. In UVD animals, the highest dose of AIDA significantly reduced SN frequency and changed its rate of compensation (P<0.001 and P<0.0001, respectively). This dose of AIDA also caused a significant reduction in YHT (P<0.005) as well as a significant change in its rate of compensation (P<0.0001). However, RHT was not significantly affected. In the labyrinthine-intact animals, AIDA infusion did not induce a UVD syndrome, nor did it significantly affect RRL. These results suggest that group I mGluRs in the ipsilateral VNC may be involved in the expression of ocular motor and some postural symptoms following UVD. Furthermore, group I mGluRs may not contribute to the resting activity of vestibular nucleus neurons. PMID- 11026730 TI - Sustained GABA-induced regulation of the inactivation of the voltage-dependent Ca2+ current in crustacean muscle fibers. AB - We describe a new form of gamma-aminobutyric acid (GABA) -mediated regulation of the inactivation and of the recovery from inactivation of the L-type Ca2+ current (I(Ca)) in crayfish muscle. GABA (1 mM) was applied during a 2-min period and the peak I(Ca) was measured using two-electrode voltage-clamp recordings 30 min after returning to the control solution. Prepulse-pulse protocols showed that the GABA mediated inhibition of I(Ca) decreased (>50%) both with increasing prepulse depolarization and as the delay between prepulse and pulse was reduced. GABA also shifted to depolarized values (>5 mV) the S-shaped plots of the peak I(Ca) evoked by a constant depolarizing pulse as a function of prepulse voltage (i.e., inactivation curves) and accelerated the recovery time from the inactivation evoked by depolarizing prepulses (>35%). The effects outlasted GABA application up to 1 h. The observed changes in inactivation properties may be of functional importance because they indicate that previous depolarization relieves the GABA induced inhibition of I(Ca), implying that this long-lasting inhibition is under the regulation of the prepulse potential and the subsequent Ca2+ entry. PMID- 11026731 TI - Three-dimensional eye-movement responses to off-vertical axis rotations in humans. AB - We recorded three-dimensional eye movements elicited by velocity steps about axes that were tilted with respect to the earth-vertical. Subjects were accelerated in 1 s from zero to 100 degrees/s, and the axis of rotation was tilted by 15 degrees, 30 degrees, 60 degrees, or 90 degrees. This stimulus induced a constant horizontal velocity component that was directed opposite to the direction of rotation, as well as a modulation of the horizontal, vertical and torsional components with the frequency of the rotation. The maximum steady-state response in the horizontal constant-velocity component was much smaller than in other species (about 6 degrees/s), reaching a maximum at a tilt angle of about 60 degrees. While the amplitude of the horizontal modulation component increased up to a tilt angle of 90 degrees (8.4 degrees/s), the vertical and torsional modulation amplitudes saturated around 60 degrees (ca. 2.5 degrees/s). At small tilt angles, the horizontal modulation component showed a small phase lag with respect to the chair position, which turned into a small phase lead at large tilt angles. The torsional component showed a phase lead that increased with increasing tilt angle. The vertical and torsional velocity modulation at large tilt angles was not predicted by a recent model of otolith-canal interaction by Merfeld. Agreement between model and experimental data could be achieved, however, by introducing a constant force along the body's z-axis to compensate for the gravitational pull on the otoliths in the head-upright position. This approach had been suggested previously to explain the direction of the perceived subjective vertical during roll under different g-levels, and produced in our model the observed vertical and torsional modulation components at large tilt angles. PMID- 11026732 TI - Multisensory information for human postural control: integrating touch and vision. AB - Despite extensive research on the influence of visual, vestibular and somatosensory information on human postural control, it remains unclear how these sensory channels are fused for self-orientation. The focus of the present study was to test whether a linear additive model could account for the fusion of touch and vision for postural control. We simultaneously manipulated visual and somatosensory (touch) stimuli in five conditions of single- and multisensory stimulation. The visual stimulus was a display of random dots projected onto a screen in front of the standing subject. The somatosensory stimulus was a rigid plate which subjects contacted lightly (<1 N of force) with their right index fingertip. In each condition, one sensory stimulus oscillated (dynamic) in the medial-lateral direction while the other stimulus was either dynamic, static or absent. The results qualitatively supported five predictions of the linear additive model in that the patterns of gain and variability across conditions were consistent with model predictions. However, a strict quantitative comparison revealed significant deviations from model predictions, indicating that the sensory fusion process clearly has nonlinear aspects. We suggest that the sensory fusion process behaved in an approximately linear fashion because the experimental paradigm tested postural control very close to the equilibrium point of vertical upright. PMID- 11026734 TI - Disconjugate memory-guided saccades to disparate targets: temporal aspects. AB - Memory-guided saccades to disparate targets (i.e., more eccentric for one eye) flashed 1 s earlier become disconjugate (i.e., of different amplitude for the two eyes) after only about 30 trials. After about 225 trials the disconjugacy persists even when the target to remember is no longer disparate. This suggests fast learning based on short-term memorization of disparity. Learning, however, fails to occur if during the training the memory delay for each trial is increased to 2 s. The purpose of the present study was to test the importance of the frequency of stimulus presentation and thereby the rate of saccades. The same memory-guided saccade paradigm was used as in the prior study and a short training period of 225 trials was applied. For each training trial, the memory delay was again 2 s, but the time allocated for fixation of the central dot and the time allocated for fixation of the remembered target in the dark was reduced to increase the frequency of saccades made. Saccades became rapidly disconjugate and their disconjugacy was retained in a subsequent neutral condition using non disparate targets. These findings indicate that stimulus frequency and thereby saccade frequency is important for disconjugate oculomotor learning based on disparity memorization. Nevertheless, additional experiments using longer memory delays of 3 s or 4 s show a definite failure of memorization and disconjugate learning. PMID- 11026733 TI - Distribution patterns of different P2x receptor phenotypes in acutely dissociated dorsal root ganglion neurons of adult rats. AB - P2x receptors may be used to detect ATP release from tissues during physiological and pathological conditions. We used whole-cell patch clamp recordings to study the expression of P2x receptor phenotypes, their distribution patterns, and their sensitivity to alphabetamATP and suramin in dorsal root ganglion (DRG) neurons acutely dissociated from adult rats. Based on the onset and decay rates of 10 microM ATP-evoked currents, we showed three types of P2x currents: fast, slow, and mixed. Each of these P2x receptor phenotypes had a distinct distribution pattern among DRG neurons. The fast P2x currents were predominantly expressed in small-diameter, isolectin-B4 (IB4)-positive, and capsaicin-sensitive DRG neurons. The slow P2x currents were expressed in both small and medium DRG neurons, and about half of them were IB4 positive. The mixed P2x currents were also expressed in both small and medium-sized DRG neurons, and most of these neurons were IB4 positive neurons. The slow and mixed P2x current groups had both capsaicin sensitive and -insensitive DRG neurons. All phenotypes revealed with 10 microM ATP could be inhibited by 30 microM suramin. All DRG neurons with fast or mixed P2x currents were also sensitive to 10 microM alphabetamATP, and alphabetamATP evoked currents similar to those induced by ATP. The group expressing slow P2x currents could be further divided into alphabetamATP-sensitive and -insensitive groups. Thus, the relationships among P2x receptor phenotypes, cell sizes, IB4 positivity, and capsaicin sensitivity are more complicated than previously thought, and different P2x receptors may be involved in both nociceptive and non nociceptive functions. PMID- 11026735 TI - Endomorphin-1 induces antinociception without immunomodulatory effects in the rat. AB - RATIONALE: Although there is evidence that central opioid receptors are involved in immunomodulation, it has been only recently that an endogenous agonist, designated endomorphin-1, possessing high selectivity and affinity for the mu opioid receptor has been identified. OBJECTIVE: The present study assesses the immunomodulatory effects of endomorphin- in the rat and provides further evaluation of the antinociceptive effects of endomorphin-1. METHODS: Rats were surgically implanted with cannulae directed at the lateral cerebral ventricle. Animals received vehicle or endomorphin-1 at doses of 31.63 or 56.23 microg (ICV) and were tested for antinociception in two different assays, the warm water tail withdrawal procedure and the hotplate assay. Additional studies assessed the effect of naltrexone on the antinociception produced by endomorphin-1 in both antinociceptive assessments. Assessments of immune status following endomorphin-1 treatment included measurements of splenic natural killer cell activity, production of interferon-y, and lymphocyte proliferative responses to mitogenic stimulation by Con-A, LPS, and the microbial superantigen, TSST-1. RESULTS: Endomorphin-1 induced significant and naltrexone reversible antinociception 30 and 60 min following drug administration, as measured by the hotplate assay and warm water tail withdrawal procedure. In marked contrast, endomorphin-1 did not produce immunomodulatory effects up to 120 min following ICV administration. CONCLUSIONS: Endomorphin-1 produces antinociception but does not induce immunomodulatory effects in the rat. These findings suggest that it is possible to develop therapeutic strategies for separating antinociception and immunomodulatory properties through the mu opioid receptor. PMID- 11026736 TI - Antidepressant-like effects of pregnancy and progesterone in Wistar rats as measured in the differential reinforcement of the low-rate 72 s task. AB - RATIONALE: In rats, several behavioral changes occurring during pregnancy could be due to the presence of progesterone; some of them may be analyzed in the differential reinforcement of the low-rate 72 s task (DRL-72 s), which is designed for testing the antidepressant profile of drugs. OBJECTIVES: The aim of the present study was to analyze the behavior of pregnant rats or ovariectomized rats receiving exogenous progesterone in the DRL-72 s task. HYPOTHESIS: During pregnancy, rats will obtain a high number of reinforcers in the DRL-72 s task. METHODS: Pregnant rats or rats after delivery were tested in the DRL-72 s task at the 3rd, 7th, 14th, 17th, and 20th days. Control rats previously trained in the DRL-72 s task were ovariectomized; after recuperation, they received saline (0.9%, i.p.), clomipramine (1.25 mg/kg, i.p.), or desipramine (2.14 mg/kg, i.p.) for 28 days, and they were tested in the DRL-72 s task. In a second series of experiments, ovariectomized rats received vehicle or progesterone (0.5, 1.0, 2.0, 4.0 mg/kg, s.c.), and they were submitted to the DRL-72 s task. Locomotion was evaluated in the open field test. RESULTS: Pregnant rats tested at the 14th and 17th day and ovariectomized rats receiving progesterone or two tricyclic antidepressants obtained a higher number of reinforcers and a cohesive rightward shift in inter-response time distributions than those rats evaluated after delivery in the DRL-72 s task. A lower locomotion was observed only at the end of pregnancy. CONCLUSIONS: Antidepressant-like effects of pregnancy and progesterone were found in Wistar rats as measured in the DRL-72 s task. PMID- 11026737 TI - Cytochrome P-450 enzymes and FMO3 contribute to the disposition of the antipsychotic drug perazine in vitro. AB - RATIONALE: Perazine (PER) is a phenothiazine antipsychotic drug frequently used in Germany that undergoes extensive metabolism. OBJECTIVES AND METHODS: To anticipate metabolic drug interactions and to explore the relevance of polymorphisms of metabolic enzymes, perazine-N-demethylation and perazine-N oxidation were investigated in vitro using human liver microsomes and cDNA expressed enzymes. RESULTS: CYP3A4 and CYP2C9 were identified as the major enzymes mediating PER-N-demethylation. At 10 microM PER, a concentration consistent with anticipated in vivo liver concentrations, CYP3A4 and CYP2C9 contributed 50% and 35%, respectively, to PER-N-demethylation. With increasing PER concentrations, contribution of CYP2C9 decreased and CYP3A4 became more important. In human liver microsomes, PER-N-demethylation was inhibited by ketoconazole (>40%) and sulfaphenazole (16%). Allelic variants of recombinant CYP2C9 showed differences in PER-N-demethylase activity. The wild type allele CYP2C9*1 was the most active variant. Maximal activities of CYP2C9*2 and CYP2C9*3 were 88% and 18%, respectively, compared to the wild type activity. Perazine-N oxidation was mainly mediated by FMO3. In the absence of NADPH, heat treatment of microsomes abolished PER-N-oxidase activity. Methimazole inhibited PER-N oxidation, while CYP specific inhibitors had no inhibitory effect. Perazine is a potent inhibitor of dextromethorphan-O-demethylase, S-mephenytoin-hydroxylase, alprazolam-4-hydroxylase, phenacetin-O-deethylase and tolbutamide-hydroxylase activity in human liver microsomes. CONCLUSIONS: Alterations in the activity of CYP3A4, CYP2C9 and FMO3 through genetic polymorphisms, enzyme induction or inhibition bear the potential to cause clinically significant changes in perazine clearance. PER may alter the clearance of coadministered compounds metabolized by CYP2D6, CYP2C19, CYP2C9, CYP3A4 and CYP1A2. PMID- 11026738 TI - Naltrexone effects on ethanol consumption and response to ethanol conditioned cues in C57BL/6 mice. AB - RATIONALE: The conditions under which naltrexone reduces ethanol consumption and its effect on behavior controlled by ethanol conditioned stimuli remain unclear. OBJECTIVES: The objectives were to determine the effects of naltrexone on ethanol consumption by C57BL/6 (B6) mice when injected subcutaneously (expt 1) or delivered by osmotic minipump (expt 2), and on ethanol conditioned cues (expt 3). METHODS: Naltrexone effects on ethanol consumption and preference were measured in a continuous access two-bottle choice paradigm in groups of mice implanted with osmotic minipumps delivering 0-3.0 mg/kg per day or injected subcutaneously with 0-6.0 mg/kg doses. Naltrexone's (0-3.0 mg/kg) effect on ethanol-conditioned cues was indexed by its effect on the expression of ethanol place conditioning (expt 3). RESULTS: Naltrexone produced a transient reduction in ethanol consumption and a consistent reduction in preference when injected; however, it had no effect on ethanol consumption or preference when delivered continuously by osmotic minipump. Naltrexone attenuated the expression of ethanol place conditioning in a U-shaped dose-response function. CONCLUSIONS: The transient reduction in ethanol consumption produced by injected naltrexone and the absence of an effect when continuously delivered confirms a report that maintaining naltrexone at steady state levels may antagonize its attenuation of ethanol consumption. The reduced expression of ethanol place conditioning in naltrexone injected mice suggests that the drug can attenuate the reinforcing effects of ethanol conditioned stimuli as was recently reported for lever responding maintained by ethanol conditioned stimuli in rats. These effects were observed at naltrexone doses with no readily apparent adverse side-effects, supporting its usefulness for treating alcoholism. PMID- 11026740 TI - Differential development of behavioral tolerance and the subsequent hedonic effects of alcohol in AA and ANA rats. AB - RATIONALE: There at least two ways in which tolerance development to alcohol's behavioral effects could interact with its subsequent intake: 1) tolerance to alcohol's reward or reinforcing effects per se could lead to increased consumption, and 2) tolerance to alcohol's aversive effects could unmask alcohol's rewarding effects. These two mechanisms may differentially interact with preexisting genetic traits underlying alcoholism. OBJECTIVES: Alcohol's subjective attributes were assessed in selectively bred AA and ANA rats after the development of tolerance to alcohol's behaviorally disruptive effects on lever press performance. METHODS: Rats were trained to press a lever under an FR30 schedule of food presentations. Group-dependent differential access to intoxicated practice, using a typical pre-post drug administration design, was utilized to promote the development of alcohol tolerance in only the group receiving intoxicated practice sessions. Subsequently, rats were trained to associate alcohol with unique place and taste stimuli in order to assess the relative changes in the approach towards, or avoidance of alcohol-related cues in each group. RESULTS: Groups of AA and ANA rats given access to intoxicated practice demonstrated tolerance development. These groups subsequently conditioned place preferences and failed to develop conditioned taste aversions to alcohol. Passive alcohol exposure in the ANA rats set the occasion for the development of a place preference and delayed taste conditioning. AA rats exposed to passive alcohol exposure failed to condition place preferences and developed rapid taste aversions. Saline control rats failed to develop tolerance or place preferences but did condition a robust alcohol-induced taste aversion. CONCLUSIONS: AA and ANA rats differ in their behavioral and pharmacokinetic response to chronic alcohol exposure. Compensatory responses interacting with approach-avoidance behaviors appear to be learned during intoxicated practice in the AA rats and during both intoxicated practice and passive exposure in the ANA rat line. PMID- 11026739 TI - The inclusion of fluoxetine into gamma-cyclodextrin increases its bioavailability: behavioral, electrophysiological and pharmacokinetic studies. AB - The inclusion of a drug into cyclodextrin generally results in the modification of its physical and chemical properties and sometimes can increase its oral bioavailability. The aim of this study was to compare the effects of the fluoxetine HCl/gamma-cyclodextrin complex to that of traditional fluoxetine HCl. In the forced swimming test in mice, fluoxetine HCl/gamma-cyclodextrin was more effective than fluoxetine HCl, the ED30s being, respectively, 9.5 and 16.9 mg/kg PO. Both compounds (10 mg/kg PO) were able to reduce the firing rate of dorsal raphe neurons in the rat. However, between-groups comparisons showed no significant differences between fluoxetine HCl treated animals and the vehicle group, while fluoxetine HCl/gamma-cyclodextrin appeared significantly more effective than vehicle from minute 25 of the measurement period. In healthy volunteers, the relative oral bioavailability, calculated as the ratio AUC 0 infinity fluoxetine HCl/gamma-cyclodextrin on AUC 0-infinity fluoxetine HCl (20 mg PO), was equal to 249.9%. The three experiments taken together suggest that the complexation of fluoxetine HCl into gamma-cyclodextrin increases its pharmacological efficacy in animals, this effect being related to an enhancement of its oral bioavailability as demonstrated in human healthy subjects. PMID- 11026741 TI - Nociceptin/orphanin FQ (N/OFQ) induces a quasi-morphine abstinence syndrome in the rat. AB - Nociceptin/orphanin FQ (N/OFQ) is a neuropeptide that exerts antiopiate effects under some circumstances, and there is evidence that it contributes to opiate tolerance. This raises the question, might N/OFQ also contribute to opiate dependence and abstinence? Twenty-two male Sprague-Dawley rats were cannulated in the third ventricle and challenged 7 days later by third ventricle injection of 50, 200 or 1,000 ng N/OFQ or saline alone. Each rat was observed under "blind" conditions for 30 min beginning 15 min after onset of the third ventricle injection. There was a significant positive linear trend of signs as a function of N/OFQ dose. Subjects receiving saline had 18.0+/-2.0 (mean+/-SEM) overall abstinence-like signs, whereas subjects receiving 50, 200 or 1000 ng N/OFQ had 35.2+/-3.6, 49.8+/-2.6 and 63.5+/-9.7 signs, respectively. In 16 additional rats, abstinence-like signs induced by 1000 ng N/OFQ were significantly attenuated by low SC doses of morphine or clonidine. These results raise the possibility that N/OFQ might contribute to opiate dependence and subsequent abstinence syndrome. On the other hand, N/OFQ over a wide dose range induced abstinence signs with similar potency in morphine dependent and non-dependent rats. PMID- 11026742 TI - Dynorphin A (2-17) attenuates the unconditioned but not the conditioned effects of opiate withdrawal in the rat. AB - OBJECTIVES: An unbiased place preference conditioning procedure was used to examine the influence of the non-opioid peptide, dynorphin A 2-17 (DYN 2-17), upon the conditioned and unconditioned effects of opiate withdrawal in the rat. METHODS: Rats were implanted SC with two pellets containing 75 mg morphine or placebo. Single-trial place conditioning sessions with saline and the opioid receptor antagonist naloxone (0.1-1.0 mg/kg; SC) commenced 4 days later. Ten minutes before SC injections, animals received an IV infusion of saline or DYN 2 17 (0.1-5.0 mg/kg). Additional groups of placebo- and morphine-pelleted animals were conditioned with saline and DYN 2-17. During each 30-min conditioning session, somatic signs of withdrawal were quantified. Tests of place conditioning were conducted in pelleted animals 24 h later. RESULTS: Naloxone produced wet-dog shakes, body weight loss, ptosis and diarrhea in morphine-pelleted animals. Morphine-pelleted animals also exhibited significant aversions for an environment previously associated with the administration of naloxone. These effects were not observed in placebo-pelleted animals. DYN 2-17 pretreatment resulted in a dose related attenuation of somatic withdrawal signs. However, conditioned place aversions were still observed in morphine-pelleted animals that had received DYN 2-17 in combination with naloxone. Furthermore, the magnitude of this effect did not differ from control animals. CONCLUSIONS: These data demonstrate that the administration of DYN 2-17 attenuates the somatic, but not the conditioned aversive effects of antagonist-precipitated withdrawal from morphine in the rat. Differential effects of this peptide in modulating the conditioned and unconditioned effects of opiate withdrawal are suggested. PMID- 11026743 TI - Synergistic enhancement of the acoustic startle reflex by dopamine D1 and 5-HT1A agonists and corresponding changes in c-Fos expression in the dorsal raphe of rats. AB - RATIONALE AND OBJECTIVES: Several studies have reported an increase in dopamine (DA)-stimulated behavioral responses after manipulations that reduce brain serotonin (5-hydroxytryptamine, 5-HT) levels. Because others have shown that systemic administration of the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) reduces 5-HT levels throughout the brain, we tested the effects of 8-OH-DPAT on the enhancement of the acoustic startle reflex by the dopamine D1 receptor agonist SKF 82958. In addition, we used the expression of the c-Fos protein as a marker of neuronal activity to assess any corresponding drug-induced changes within the dorsal raphe (DR). METHODS AND RESULTS: Male Sprague-Dawley rats pretreated (10 min) with 8-OH-DPAT (0.5 mg/kg) showed a marked potentiation of the enhancement of startle by SKF 82958 (0.1 mg/kg). Furthermore, SKF 82958 produced a dramatic induction of c-Fos in the DR, an effect that was blocked by 8-OH-DPAT. Double-labeling immunohistochemistry for c Fos and 5-HT showed that SKF 82958-induced expression of c-Fos, and its blockade by 8-OH-DPAT, occurred in a percentage of 5-HT-containing cells of the DR. CONCLUSIONS: These data suggest the possibility that inhibition of the DR by 8-OH DPAT mediates the potentiation of startle by SKF 82958, perhaps through a reduction in 5-HT release in the striatum. Such an interpretation is consistent with the hypothesis of an inhibitory role of the 5-HT system on DA-mediated behaviors. PMID- 11026744 TI - Effects of caffeine on sensorimotor gating of the startle reflex in normal control subjects: impact of caffeine intake and withdrawal. AB - RATIONALE: Prepulse inhibition (PPI), a cross-species measure of sensorimotor gating, is impaired in certain neuropsychiatric disorders. This study was designed to assess caffeine effects on PPI in normal humans, as part of an effort to understand cross-species differences and similarities in the neurochemical regulation of PPI. METHODS: Startle was measured during a screening session; 7 days later, subjects were retested after placebo or caffeine (200 mg; double blind design). Subjects were characterized as low versus high caffeine drinkers based on established scales (range 11-628 mg/day), and either maintained ad libitum caffeine intake (Ad lib study; n=18) or refrained from caffeine consumption for > or =15 h prior to testing (Withdrawal study; n=12). Autonomic and self-rating measures, acoustic and tactile startle, and unimodal and cross modal PPI, were measured in divided sessions for 3 h post-treatment. RESULTS: There were significant effects of caffeine and/or caffeine withdrawal on several self-rating and autonomic measures, and on startle reflex habituation, but not on acoustic or tactile startle magnitude or PPI. Difference scores of startle data from screening versus test days revealed no group effects on startle magnitude, but PPI difference scores revealed that caffeine had opposite effects on low versus high caffeine drinkers (means=57 versus 258 mg/day) in the two withdrawal states. In the absence of withdrawal, caffeine reduced PPI in heavy caffeine drinkers; during withdrawal, caffeine increased PPI in heavy caffeine drinkers. The opposite pattern was evident in low caffeine drinkers. CONCLUSIONS: While a physiologically active dose of caffeine has no simple effects on PPI in normal humans, both withdrawal states and normal levels of caffeine consumption may be important factors in understanding this drug's effects on sensorimotor gating. PMID- 11026745 TI - Effects of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) on performance on two operant timing schedules. AB - RATIONALE: Previous experiments have shown that the disruptive effect of central 5-HT depletion on interval timing behaviour is critically dependent upon the particular timing schedule used. However, it is not known how acute disruption of 5-HTergic function brought about by drugs acting at 5-HT receptors affects timing. OBJECTIVE: To examine the effects of the 5-HT1A receptor agonist 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) on performance on two quantitative timing schedules, a free-operant schedule in which rats were trained to distribute their responses differentially between two levers during the course of a 50-s trial (free-operant psychophysical procedure) and a discrete-trials schedule in which rats were trained to discriminate the durations of light stimuli (interval bisection task). METHODS: In experiment 1, rats were trained under the free-operant psychophysical procedure to respond on two levers (A and B) in 50-s trials in which reinforcement was provided intermittently for responding on A in the first half, and B in the second half, of the trial. For one group, repetitive switching between levers was permitted; for another group, it was prevented. In experiment 2, rats were trained to press lever A after a 2-s stimulus and lever B after an 8-s stimulus, and were then tested with stimuli of intermediate durations. For one group, a 'poke response' (depression of a central tray flap) was required after stimulus presentation to effect lever presentation; for the other group this requirement did not operate. In both experiments, quantitative indices of timing were derived from the psychophysical functions (%B responding vs time). RESULTS: In experiment 1, 8-OH-DPAT (25, 50, 100 and 200 microg kg(-1) s.c.) displaced the psychophysical curve to the left in both versions of the schedule. In experiment 2, 8-OH-DPAT increased the Weber fraction in both versions of the task without displacing the curve. CONCLUSIONS: These results show that 8-OH-DPAT disrupts timing behaviour. The results of experiment 1 are consistent with the proposal that 5-HTergic mechanisms help to regulate the period of the hypothetical pacemaker. However, the results of experiment 2 do not support this suggestion. Taken together, the results support the notion that different neural mechanisms may be involved in timing tasks involving temporal distribution of responding and discrimination of the durations of exteroceptive stimuli. PMID- 11026746 TI - Nicotine self-administration in rats: estrous cycle effects, sex differences and nicotinic receptor binding. AB - RATIONALE: Research on smoking behavior and responsiveness to nicotine suggests that nicotine's effects may depend on the sex of the organism. OBJECTIVE: The present study addressed four questions: 1) Will female rats self-administer nicotine? 2) Does self-administration by females vary as a function of estrous cycle? 3) Does self-administration by females differ from that of males? 4) Does self-administration of nicotine result in up-regulation of nicotinic receptor binding and are these changes similar in males and females? METHODS: Male and female Sprague-Dawley rats were allowed to self-administer nicotine at one of four doses (0.02-0.09 mg/kg, free base) on both fixed and progressive ratio schedules of reinforcement. RESULTS: Females acquired nicotine self administration across the entire range of doses. Acquisition of self administration at the lowest dose was faster in females than males. However, few sex differences were found in the number of active responses, number of infusions, or total intake of nicotine during stable fixed ratio self administration. In contrast, females reached higher break points on a progressive ratio. For both schedules, females had shorter latencies to earn their first infusion of each session and demonstrated higher rates of both inactive and timeout responding. There was no effect of estrous cycle on self-administration during either fixed or progressive ratio sessions. Self-administered nicotine resulted in average arterial plasma nicotine levels between 53 and 193 ng/ml and left hemi-brain levels between 174 and 655 ng/g, depending on dose. Nicotine self administration produced similar up-regulation of nicotinic receptor binding sites in males and females, as reflected by increased right hemi-brain binding of [3H] epibatidine, when compared to the brains of untreated control rats. CONCLUSIONS: These results suggest that while males and females may regulate their intake of nicotine similarly under limited access conditions, the motivation to obtain nicotine is higher in females. PMID- 11026747 TI - Repeated pretreatment with amphetamine sensitizes increases in cortical acetylcholine release. AB - RATIONALE: Previous studies on the attentional effects of repeated psychostimulant administration in rats suggested the possibility that these effects are mediated via increases in the efficacy of psychostimulants to stimulate cortical acetylcholine (ACh) release. Furthermore, neurochemical data have raised the possibility that increases in nucleus accumbens (NAC) dopamine (DA) release trans-synaptically increase the excitability of basal forebrain corticopetal cholinergic projections, thereby supporting speculations about relationships between the effects of repeated psychostimulant administration on NAC DA and cortical ACh release. OBJECTIVES: To determine whether repeated exposure to amphetamine would potentiate the stimulating effects of the drug on cortical ACh and NAC DA efflux. METHODS: Rats were implanted with microdialysis guide cannula in the medial prefrontal cortex and the shell region of the ipsilateral NAC. Amphetamine (2.0 mg/kg i.p.) or saline (0.9%) was administered every other day for 10 days, for a total of five injections. ACh and DA efflux and locomotor activity were measured on the day of the first and last injections of this pretreatment regimen. All animals were retested following a challenge dose of amphetamine (2.0 mg/kg i.p.) given 10 and 19 days after the last pretreatment injection. RESULTS: The initial injections of amphetamine stimulated ACh and DA efflux and locomotor behavior in both groups. The pretreatment with amphetamine potentiated the ability of the drug to stimulate cortical ACh efflux on day 19 of the withdrawal period. The pretreatment with amphetamine also increased the effects of the challenge dose on motoric activity on day 10. Pretreatment with amphetamine did not result in a significant augmentation of the amphetamine-induced increase in DA efflux in the NAC. CONCLUSIONS: Pretreatment with amphetamine sensitizes the ability of amphetamine to stimulate cortical ACh efflux. These results support the hypothesis that sensitized release of cortical ACh mediated the previously observed hyperattentional impairments in amphetamine pretreated rats. Sensitized cortical ACh release following repeated exposure to psychostimulants may mediate the overprocessing of addictive drug-related stimuli, thus contributing to repeated compulsive addictive drug use. PMID- 11026748 TI - The dose-dependent cognitive effects of acute administration of Ginkgo biloba to healthy young volunteers. AB - RATIONALE: Chronic administration of extracts from the leaves of the tree Ginkgo biloba is known to improve aspects of cognitive performance. However, little is known about the effects of acute doses of Ginkgo on coherent cognitive domains. Recent factor analysis of test measures from subtasks of the Cognitive Drug Research (CDR) computerised assessment battery has revealed that four primary cognitive 'factors' corresponding to speed of attention, accuracy of attention, speed of memory and quality of memory can be useful to describe cognitive function changes. OBJECTIVE: The present study aimed at assessing whether acute administration of Ginkgo biloba had any consistent effect on the four CDR factors. METHODS: The study utilised a placebo-controlled, multi-dose, double blind, balanced, crossover design. Twenty participants received 120 mg, 240 mg and 360 mg of a standardised extract of Ginkgo (GK501, Pharmaton, SA) or a matching placebo. Cognitive performance was assessed using the CDR computerised test battery immediately prior to dosing and at 1, 2.5, 4 and 6 h thereafter. The primary outcome measures were the four aspects of cognitive performance, which have previously been derived by factor analysis of CDR subtests. RESULTS: Compared with the placebo, administration of Ginkgo produced a number of significant changes on the performance measures. The most striking of these was a dose-dependent improvement of the 'speed of attention' factor following both 240 mg and 360 mg of the extract, which was evident at 2.5 h and was still present at 6 h. Additionally, there were a number of time- and dose-specific changes (both positive and negative) in performance of the other factors. CONCLUSIONS: We conclude that acute administration of Ginkgo biloba is capable of producing a sustained improvement in attention in healthy young volunteers. PMID- 11026749 TI - A PET study of brain 5-HT2 receptors and their correlation with platelet 5-HT2 receptors in healthy humans. AB - RATIONALE: Platelets share many properties with brain serotonergic neurons such as active 5-hydroxytryptamine (5-HT) transport, 5-HT2 receptors, and mitochondrial monoamine oxidase. OBJECTIVES: We measured brain 5-HT2 receptors and platelet 5-HT2 receptors in healthy volunteers to determine if there was any correlation between the two measures. METHODS: Ten healthy volunteers with no lifetime history of psychiatric illness or family history in first-degree relatives were recruited. 5-HT2 receptor binding was determined for each subject with positron emission tomography and [18F]setoperone scan in the brain and with 3H-LSD binding in platelets. RESULTS: We found no significant correlation between 5-HT2 binding potential (BP) in platelets (Bmax/Kd) and a semiquantitative estimate of 5-HT2 BP in frontal, parietal, and temporal cortical regions. SPM voxel based analysis also showed no significant correlation between the 5-HT2 BP in platelets and in the brains of the study subjects. CONCLUSIONS: Brain 5-HT2 receptor binding was not significantly correlated to platelet 3H-LSD binding in healthy subjects. This raises questions about the validity of generalizing findings from platelet studies to 5-HT neurons in the brain. PMID- 11026750 TI - Appetitve effects of drug cues modelled by pictures of the intake ritual: generality of cue-modulated startle examined with inpatient alcoholics. AB - RATIONALE: A cue-modulated startle test recently confirmed that smoking cues in smokers may not be withdrawal-like and aversive as traditionally believed. OBJECTIVE: Analogous tests were applied to alcohol cues in inpatient alcoholics. METHODS: Twenty-six withdrawn alcoholics (18 men) were examined. Alcohol-related pictures were compared to standardised pleasant, neutral or unpleasant control scenes using an acoustic startle test and measures of pleasure, arousal and desire for alcohol. RESULTS: Pictures depicting preparation for drinking (cues) were different from unpleasant control pictures and similar to pleasant pictures but only on the startle test; no differential effects were found for alcohol craving and mixed motivational effects were reported subjectively. The effects were not due to arousal and control pictures depicting post-drinking events showed less effect than the alcohol cues. CONCLUSIONS: New techniques of measuring drug cue motivation indicate that drug cues may have incentive properties in individuals in treatment for alcoholism. PMID- 11026751 TI - Central nervous system structures connected with the endocrine glands. findings obtained with the viral transneuronal tracing technique. AB - This review is a summary of recent neuromorphological observations on the existence of multisynaptic neural pathways between the endocrine glands and the central nervous system (CNS) and its structures involved in this pathway. Introduction of the viral transneuronal tracing technique has made possible investigations of multisynaptic connections. The utility of this approach is based on the ability of the neurotropic virus to invade and replicate in neurons, and then gradually infect synaptically linked second-order, third-order. etc. neurons. Injecting the virus into the endocrine glands, this technique was used to identify cell groups in the spinal cord and in the brain which are connected with the adrenal gland, the gonads and the pancreas. Injection of the virus into these organs resulted in viral labeling of neurons in practically identical structures of the CNS including the intermediolateral cell column of the spinal cord, the vagal nuclei and certain other cell groups in the brain stem. In the hypothalamus the most intensive labeling was in the parvocellular part of the paraventricular nucleus and in the telencephalon labeled nerve cells were detected in the amygdala, the bed nucleus of the stria terminalis and in the preoptic area. It is known that the labeled CNS structures are members of descending pathways arising from the hypothalamic paraventricular nucleus or from other cell groups and terminating on neurons of the vagal nuclei and the intermediolateral cell column of the spinal cord. Experimental data support the view that the CNS structures and pathways connected with the endocrine glands are involved in the neural control of these organs. PMID- 11026752 TI - Copper, zinc and magnesium levels in fibrocalculous pancreatic diabetes. AB - Alterations in trace elements and mineral homeostasis have been documented both in insulin-dependent diabetes mellitus and non-insulin dependent diabetes mellitus. No data are available about trace elements in fibrocalculous pancreatic diabetes, a unique form of secondary diabetes mellitus. This study evaluated the plasma concentrations of copper, zinc and magnesium in this form of diabetes. Twenty-five patients (9 men and 16 women) with fibrocalculous pancreatic diabetes and 25 healthy non-diabetic subjects (16 men and 9 women) were studied. Patients with overt nephropathy were excluded. Plasma copper, zinc, and magnesium levels were analyzed using a GBC 902 double beam absorption spectrophotometer. The effect of glycemic control, microalbuminuria, sex and modality of treatment received on the plasma levels of copper, zinc and magnesium was assessed. Results of the study revealed that plasma copper, zinc, and magnesium levels were comparable between patients with fibrocalculous pancreatic diabetes and control subjects. Plasma copper levels were significantly higher in patients with controlled diabetes (16.15 +/- 0.67 micromol L(-1)) as compared to those with uncontrolled diabetes (13.75 +/- 0.61 micromol L(-1)) and healthy controls (13.91 +/- 0.55 micromol L(-1)). This merits further investigation. Microalbuminuria, modality of treatment received and sex did not influence the levels of these elements in fibrocalculous pancreatic diabetes. PMID- 11026753 TI - Liver transplantation and diabetes mellitus. AB - Post-transplant diabetes mellitus (PTDM) is a common complication after orthotopic liver transplantation (oLT). In our study, we investigated the prevalence and risk factors one year after transplantation in 618 patients who underwent oLT between 1990 and 1996 in a single center. The influence of steroid medication and hepatitis B or C (HBV/HCV) was also studied. Before oLT 66 of the 618 patients were diabetic. After transplantation 37 of these 66 (56%) patients showed no further signs of DM. Of the 552 patients without DM before transplantation 39 (7.2%) developed new onset PTDM. There was no influence of steroid medication on the presence of PTDM (steroids 10.4% PTDM, no steroids 12.5% PTDM). In addition we found no influence of HBV or HCV-infection on PTDM development. Analysis for risk factors showed no significant influence of the diagnosis leading to oLT, of FK506 or Cyclosorin A medication, age, gender or Child-Pugh class. Five year patient survival was not influenced by the presence of PTDM, especially patients with a preexisting DM showed no reduced survival. However, a subgroup of patients with new onset insulin-requiring PTDM showed significantly reduced 5 year survival (p<0.05). In conclusion we found new onset PTDM in 7.2% of patients undergoing oLT one year after the operation. On the other hand in more than 50% of patients with preexisting DM, the disease was no longer present post-transplant. This could be an indication that DM is dependent on liver function in these patients. Patients with preexisting DM should not be excluded from transplantation if indicated. Development of new onset insulin requiring PTDM could be an important prognostic factor for patient survival after oLT. Further investigations are necessary to evaluate the prognostic meaning of PTDM and the pathophysiologic mechanisms. PMID- 11026754 TI - Non-invasive blood glucose monitoring by means of near infrared spectroscopy: investigation of long-term accuracy and stability. AB - Modern strategies in the treatment of Diabetes mellitus require frequent blood glucose tests. Near-infrared spectroscopy [NIR] is considered to be a method for non-invasive blood glucose testing. The present study evaluated the long-term accuracy and stability of the method. Spectra of 10 patients were recorded. An individual calibration models were calculated for each patient from spectra which were recorded at the beginning of the investigation. These models were then applied to calculate blood glucose values from spectra which were recorded 84 to 169 days later. The long-term accuracy and stability of the calibration models expressed as root mean squared error of prediction [RMSP] varied from 3.1 to 35.9 mmol/l and the error grid analysis [EGA] from 13 to 92% clinically acceptable values in zones A and B. The results of the investigation indicate that long-term monitoring of blood glocose by means of NIR-spectroscopy and individual calibration models over a period of 3 to 4 months is basically possible once long term stability of the method will be improved. Further technological progress and a better understanding of the underlying physiological processes is necessary to achieve a level of accuracy, reliability and stability of the method as required for clinical application. PMID- 11026755 TI - Sex steroids and bone metabolism: comparison of in vitro effects of 17beta estradiol and testosterone on human osteosarcoma cell lines of various gender and differentiation. AB - Although numerous clinical studies have demonstrated the beneficial effect of preventing postmenopausal bone loss in elder women by long-term estrogen administration, effects of estrogen at the cellular level still remain unclear. Efforts to determine the precise role of bone cells in estrogen-mediated pathways are often hampered by the lack of suitable cell culture models. Presuming that sex steroids have a direct, stimulating effect on bone cells in vitro, we investigated the influence of 17beta-estradiol, testosterone and 1,25(OH)2D, on cell proliferation and differentiation using four established human osteosarcoma (HOS) cell lines of different gender of the donors (male origin: MG 63, HOS 58; female origin: SaOS 2, TE 85). These cell lines are believed to represent different stages of osteogenic maturation. Thus, the aim of this study was to clarify if possible responses to sex steroids are related to gender or osteogenic commitment of the individual cell culture. HOS cells were cultured in six-well plates and underwent hormone treatment (1 nM and 10 nM 17beta-estradiol. 0.1 nM and I nM testosterone and 1 microM 1,25(OH)2D3) for 48 h hours. Cell proliferation was determined by measuring total cell numbers. Cell function was studied by measuring alkaline phosphatase activity and secreted osteocalcin. In this study, estrogen significantly increased proliferation of both one male (MG 63) and one female (SaOS 2) cell line, but decreased proliferation of the female HOS TE 85 cell line significantly. Testosterone treatment had a positive effect on proliferation of only one female cell line (SaOS 2). A significant increase of alkaline phosphatase activity in SaOS 2 and HOS 58 cells and of osteocalcin levels in SaOS 2 cells was detected following estrogen treatment. Administration of 1.25(OH)2D3 was followed by an increased cell proliferation in HOS 58, MG 63 and SaOS 2. Significant gender-related differences could not be demonstrated. In conclusion, response to hormonal treatment with sex steroids is not related to the gender of the osteosarcoma cell line, but rather depends on its osteoblastic commitment. PMID- 11026756 TI - Critical role of nitric oxide in expression of porcine LH receptor at transcription and post-transcription levels. AB - The present study was performed to clarify the involvement of nitric oxide (NO) in the expression of LH receptor in porcine granulosa cells. The granulosa cells, prepared from porcine ovarian follicles, were developed in the presence of FSH. LH receptor mRNA was induced to reach a maximal level after a 24-h culture with FSH, as determined by semi-quantitative reverse transcriptase-PCR (RT-PCR). In our previous report (Nishida et al., 2000), we found that NO was released from granulosa cells after a 40-48 h culture with FSH. When 200 microM NO scavenger was added to cultures before the NO release (30 h), the content of LH receptor on the cells decreased to 28% that of the control. However, the receptor content was not influenced by addition of NO scavenger at 46 h, or by 50 microM NO donor at 30 or 46 h. During transformation of mature granulosa cells to luteal cells, LH receptor mRNA was induced after a 24-h culture with LH, which induction was inhibited by removal of endogenous NO. However, the expression was not influenced by addition of either NO scavenger at 46 h or by NO donor. During the period of these treatments, cellular DNA contents were constant. Consequently, the transient generation of NO may play a critical role in expression of the LH receptor at transcription and post-transcription levels. PMID- 11026757 TI - Anorexia nervosa in congenital adrenal hyperplasia: long-term follow-up of 4 cases. AB - Studies which evaluate the psychosocial development and integration of adult female congenital adrenal hyperplasia (CAH) patients are rare but show that patients with the salt wasting form are significantly more virilized and more frequently single and childless. Major complaints are irregular menstruation, hirsutism, acne, obesity, deep voice, and cushingoid features. Surprisingly, a higher prevalence of psychosomatic disorders has not yet been described. Since anorexia nervosa (AN) has not yet been described in patients with CAH, we here report 4 cases of female CAH patients who developed AN during adolescence. Diagnosis of CAH was made between the age of 10 days and 3 years. Three patients suffer from the simple-virilizing form of CAH, one patient has a mild salt wasting CAH. Genital malformation varied from Prader stage II to IV. All 4 patients were compound heterozygotes for mutations/deletions of the CYP21B gene. Control of substitution therapy consisting of hydrocortisone and fluorocortisone was good. AN developed at ages 12, 13, 17, and 21 years (ICD 10 criteria for AN are BMI below 17.5 kg/m2, deliberate weight loss, body image disturbance, and primary or secondary amenorrhoea). Diagnosis of AN was established by psychiatrists and/or psychologists. All four patients showed an impressive and deliberate weight loss between 13 and 20 kg within 6 months, had primary or secondary amenorrhoea, and presented with BMI between 13 and 17.9 kg/m2. All patients received psychological treatment and recovered. However, one patient had a severe relapse of AN. Two patients are now married and one has a healthy son. These cases demonstrate that the diagnosis of CAH is compatible with the development of AN and illustrate the importance of providing treatment for CAH patients that encompasses not only medical but also psychological and social care. PMID- 11026758 TI - Short and long-term effects of streptozotocin on dietary cholesterol absorption, plasma lipoproteins and liver lipoprotein receptors in RICO rats. AB - Adult male genetically hypercholesterolemic RICO rats were studied 6 and 28 days after streptozotocin (STZ) administration together with untreated RICO controls. The absorption coefficient of dietary cholesterol was determined using dual isotope blood ratio method. Plasma lipoproteins as well as fecal neutral sterols and bile acids were analysed at both experimental times. Liver lipid parameters were measured and lipoprotein receptors (LDLr, SR-BI and HB2) were assayed by immunodetection. Six days after STZ administration, dietary cholesterol absorption was more efficient (+49%) in treated rats than in controls, and stayed higher (+68%) in the diabetic rats sacrificed at day 28. Fecal neutral sterol elimination decreased soon after STZ administration (by 35% at day 6), due to a higher cholesterol absorption coefficient, then increased to control level at day 28, due to installed diabetes-induced hyperphagia. Comparison of the lipoprotein profiles indicated that the concentration of HDL1. which is typically high in control Rico rats, fell significantly in diabetic rats at both experimental times, whereas that of HDL2 increased only at day 28. In diabetic rats, an early and strong enhancement of the hepatic expression of SR-BI appeared at day 6 (+415%) and persisted at day 28, but at a lesser extent (+85%). The expression of LDLr and HB2 was unchanged at day 6, but was significantly modified at day 28 (+140% for LDLr and -50% for HB2). These data show that streptozotocin-induced diabetes in Rico rats results in modifications of the expression of liver lipoprotein receptors which can contribute to alterations of the lipoprotein profile. PMID- 11026759 TI - Magnetic resonance and endoscopic retrograde cholangiopancreatography. PMID- 11026760 TI - A comparative study of magnetic resonance cholangiography and direct cholangiography. AB - OBJECTIVE: Magnetic resonance cholangiopancreatography (MRCP) is a rapidly developing method for the noninvasive assessment of the biliary tree and pancreatic duct that obviates the need for contrast medium. We describe our experience with this new diagnostic imaging method in patients with obstruction of the biliary tree. We assessed both the location and cause of obstruction, and compared the results with direct cholangiography. METHODS: Between 1997 and 1998, 81 patients underwent MRCP at our facility. Two different image acquisition protocols (half-Fourier acquisition single-shot turbo spin-echo -HASTE- and rapid acquisition with relaxation enhancement -RARE-) for T2-weighted turbo spin echo sequences as well as cross-sectional fast multiplanar gradient-echo pulse (T1 weighted FL2D) and T2-weighted fast spin echo (T2 TSE) sequences were used. All patients underwent direct (either percutaneous or endoscopic retrograde) cholangiography or surgery for confirmation and/or treatment. The images obtained with MRCP were evaluated by two radiologists with expertise in biliary tree imaging who were unaware of the patient's clinical characteristics, and their diagnostic interpretations were compared with the findings obtained upon surgery or direct cholangiography. RESULTS: The sensitivity and specificity of MRCP in ruling out pathologies and detecting the presence of dilatation of the biliary tree were 100%. In assessing the level of the obstruction, sensitivity and specificity varied with location (intrahepatic/hilar, suprapancreatic, intrapancreatic or ampullary). Sensitivity in these locations was 100, 92, 69 and 86%, respectively, whereas specificity was 100, 94, 92 and 91%, respectively. In determining the cause of the obstruction, the results were variable depending on the cause: choledocholithiasis (sensitivity, 89%; specificity, 90%) malignant obstruction (sensitivity, 92%; specificity, 88%), benign stricture (sensitivity, 63%; specificity, 90%), and chronic pancreatitis (sensitivity, 50%; specificity, 99%). CONCLUSIONS: MRCP offered high diagnostic accuracy in the assessment of the occurrence and location of biliary obstruction. Sensitivity and specificity in establishing the cause varied, and were highest for choledocholithiasis and malignant obstruction. MRCP may be used instead of ERCP, which may then be reserved for patients who are likely to require surgery. PMID- 11026761 TI - Quantitative study of metals in bile from patients with cholelithiasis. AB - OBJECTIVE: Biliary lithiasis is a multifactorial phenomenon that is decisively influenced by the composition of bile. We analyzed the presence of eight metals in bile and compared their concentrations in healthy persons and patients with cholelithiasis. METHODS: We studied bile from 119 patients who underwent cholecystectomy because of symptomatic cholelithiasis, and from 25 control subjects in whom the gallbladder was removed for reasons other than cholelithiasis. Metal concentrations were analyzed by atomic absorption spectrophotometry. The subjects were divided into subgroups according to age, sex and type of stone. RESULTS: Bile from patients with cholelithiasis contained significantly less of the essential element magnesium (Mg) and the toxic element lead (Pb) than bile from control subjects. Calcium (Ca) and strontium (Sr) concentrations were also lower in patients with gallstones than in the control group, although the differences were not significant. CONCLUSIONS: Biliary concentrations of Mg and Pb were significantly lower in patients with cholelithiasis than in the control group. The biliary excretion of Ca and Sr was lower in patients than in controls, although the differences were not statistically significant. PMID- 11026762 TI - Clinical characteristics of nonneoplastic cavernomatous transformation of the portal vein at a Gastroenterology Service in Spain. AB - OBJECTIVE: To identify predisposing factors, clinical characteristics and effective treatment in patients with nonneoplastic cavernomatous transformation of the portal vein in our gastroenterology service. METHODS: We retrospectively reviewed the clinical records of 2,201 patients diagnosed as having portal hypertension (2,165 with cirrhosis and 36 with noncirrhotic portal vein hypertension) during the period from 1977 to 1998. The diagnosis of cavernomatous transformation was confirmed with angiographic or Doppler echographic studies, or both. RESULTS: Thirteen patients (6 males, 7 females, age range 8 to 69 years) with cavernomatous transformation were found. Predisposing factors were omphalitis (1), echinococcal cyst (1), major abdominal surgery (3), liver cirrhosis (3), Sjogren syndrome (1), and no apparent cause (4). Eleven of the 13 patients had upper digestive tract bleeding from varices, 9 had splenomegaly, and 2 had cirrhotic decompensation. Splenectomy was done in 3 patients on admission, and in 5 patient shunts were used (splenorenal in 4, mesenteroatrial in 1) because of repeated bleeding. CONCLUSIONS: Of the patients with noncirrhotic portal hypertension, 27.7% had nontumoral cavernomatous transformation of the portal vein. Previous abdominal surgery was the most frequent predisposing factor; the 2 cases of echinococcal liver disease and Sjogren disease were exceptional. Age younger than 30 years, bleeding esophageal varices and splenomegaly were the most frequent clinical features. Portosystemic shunt was the only effective treatment alternative in these patients. PMID- 11026764 TI - [Conservative management of severe esophageal complication with fibrin glue (tissucol)]. PMID- 11026763 TI - Risk factors for hepatocellular carcinoma in patients with liver cirrhosis. AB - OBJECTIVE: Although cirrhosis is known to predispose toward hepatocellular carcinoma (HCC), there is no agreement on the factors that can influence the risk for HCC in patients with cirrhosis. This study was designed to identify differences in cirrhosis-related risk factors for developing HCC in relation to epidemiological characteristics, stage of the disease and etiology. METHODS: 512 patients from southwestern Spain with Child-Pugh stage A or B cirrhosis were examined periodically by ultrasonography, and alpha-fetoprotein (AFP) concentration was measured. RESULTS: The average length of follow-up was 37 months. A total of 52 cases of HCC were detected, which represented a risk of 17% after 5 years of follow-up. The Cox model showed that the risk of HCC increased by 8% per year of increasing age. Male sex (relative risk: 3.4), hepatitis C virus infection (relative risk: 4.6), hepatitis B virus infection (relative risk: 2.9) and AFP levels higher than 15 ng/ml (relative risk: 2.5) were also shown to be risk factors. Among alcoholic patients, only age (risk increased by 15% per year), and hepatitis C virus infection (relative risk: 5.4) were risk factors for HCC. However, in patients infected by hepatitis C virus, the main risk factors were age (relative risk increased by 8% per year), male sex (relative risk: 3.9), co-infection with hepatitis B virus (relative risk: 4.9), and increased AFP (relative risk: 2.8). Of the patients with HCC, 71% were infected with hepatitis C virus. Alcoholism, Child-Pugh stage and duration of cirrhosis did not increase the risk of the appearance of HCC. CONCLUSIONS: The risk of HCC increased to 17% after 5 years of follow-up in patients with Child-Pugh stage A or B cirrhosis. Hepatitis C virus infection was the main risk factor in patients with cirrhosis. Other risk factors were age, male sex, hepatitis B virus infection and altered AFP level. PMID- 11026765 TI - [Severe hepatitis with cytolysis pattern caused by amoxicillin-clavulanate]. PMID- 11026766 TI - [Hemochromatosis and hilar cholangiocarcinoma: report of a case]. PMID- 11026767 TI - [Obstructive jaundice caused by pancreatic head tumor secondary to IgG multiple myeloma]. PMID- 11026768 TI - [Intestinal necrosis as presentation form of mesenteric cyst]. PMID- 11026769 TI - A vision for health system reform in the Americas. PMID- 11026770 TI - [The context of health care reforms]. AB - In Latin America, health sector reforms have gone hand in hand with social and economic trends during the latter half of the twentieth century and have reflected the particular concept of "development" that has been in vogue at different times. Economic stagnation and increased social spending, both hallmarks of the 1960s, led to the decline of the "import substitution" development model, which had prevailed since the beginning of the century, and slowly gave way in the 1980s to the "globalization" model. From the earlier model, a transition took place toward a restructuring of production and a series of economic adjustment policies that led, ironically, to an increase in poverty in Latin America. Implementation of the new model has occurred in two phases. The first, known as the "social reform" or "first generation" phase, sprang from the notion that poverty is the sum of a number of material shortages that can be corrected through an equitable redistribution of a fixed volume of goods belonging to society. This conceptual framework, which was completely devoid of all historical linkages and separated from economic policy, led to social policies whose entire purpose was to mitigate poverty through subsidies targeting the poorest persons in the society. In the second phase of the globalization model, which arose in the 1990s and became known as the "second generation" or "postadjustment" phase, new economic rules came into play that were based primarily on international competition, efficiency in production, and openness and fairness in the capital markets. And if during the initial stage the conceptual strategy behind all social policy was to fight poverty, in the second stage the strategy became one of achieving equity, which was no longer interpreted as the even distribution of a fixed volume of capital goods, but as the sustained provision of greater and better opportunities for all. Having grown accustomed to the protectionism inherent in the earlier development model, Latin American societies today feel threatened by a new model that offers them no social safety net. The feasibility of economic and social reform policies during the second phase, which reflect the demands of a "globalized" world, thus depends on the ability to overcome people's lack of trust and to garner the support of a political, social, and institutional majority. PMID- 11026771 TI - [Reform of health systems in Latin America and the Caribbean: situation and trends]. AB - In the early 1990s, most Latin American and Caribbean countries were beginning, or planning to begin, health sector reform processes. This paper presents the status and trends of health sector reform at the end of the 1990s. The authors relied on information in 20 health system and services profiles completed by the Pan American Health Organization between August 1998 and October 1999. The analysis, which follows a methodology that had been applied earlier, was organized on two different levels: (1) monitoring the reform processes (dynamics and content) and (2) evaluating their outcomes. In looking at the dynamics of the reform processes, the article examines the context in which they take place and the actors involved in their different phases: inception, design and negotiation, implementation, and evaluation. The description and analysis of the contents of health sector reform initiatives are organized into 12 broad thematic areas. Outcomes evaluation was only possible in the eight countries that provided enough pertinent information, and should be viewed as preliminary. Nevertheless, the article does present detailed information on the outcomes of health sector reform in terms of equity, effectiveness and quality, efficiency, sustainability, and societal participation and control. The article also discusses the potential causes and determining factors for the observed outcomes. PMID- 11026772 TI - [Strengthening of the steering role of health++ authorities in health care reforms]. AB - Strengthening the ability of health authorities to provide leadership and guidance, now and in the future, is an important issue within the context of health sector reform. It means, among other things, redefining the role of health in light of leading social and economic trends seen in the world at the beginning of the 21st century, increasing participation in health by nongovernmental entities, moving toward participatory democracy in many countries, and modifying concepts of what is considered "public" and "private." Within this scenario, it is necessary to redirect the role of the health sector toward coordinating the mobilization of national resources, on a multisectoral scale, in order to improve equity and social well-being and to channel the limited available resources to the most disadvantaged groups in society. The liberalization of the production and distribution of health-related goods and services, including insurance, challenges the exercise of authority in the area of health. Furthermore, the formation of regional economic blocks and the enormous weight wielded by multinational companies in the areas of pharmaceuticals and other medical supplies and technologies are forcing the health sector to seek ways of harmonizing health legislation and international negotiations. According to many experts, all of these demands surpass the ability of Latin American ministries of health to effectively respond, given most countries' current organizational, legal, and political conditions and technical infrastructure. The countries of the Americas must make it a priority to strengthen their health officials' ability to provide leadership and guidance in order to meet present and future challenges. PMID- 11026773 TI - [Insurance and coverage: two critical topics in health care reforms]. AB - The goal of health for all in the year 2000, which was established at Alma Ata more than two decades ago, has led countries in Latin America and the Caribbean to adopt health sector reforms aimed at extending health coverage to each and every individual citizen. Whereas much has come about as a result of reform policies in the way of theory and legislation, in practice the goals that were established are far from attained, and many countries show large gaps in theoretical coverage on the one hand, and true coverage on the other. This is largely due to organizational features and other "endogenous" characteristics of the various countries' health systems, as well as to "exogenous" factors in the political, macroeconomic, social, epidemiologic, and cultural spheres. This documents takes a close look at the different types of health systems that are currently operating in countries of the Region and their impact on sources of health insurance and health coverage for individuals living in those countries. The end of the article focuses on the different strategies adopted by the countries in an effort to extend health coverage, which in some cases involve policies targeting the most vulnerable social groups. PMID- 11026774 TI - [Impact of health care reform on human resources and employment management]. AB - According to those in charge of health sector reform, human resources are the key component of health sector reform processes and offer health services their greatest competitive advantage. With the help of the Observatory for Human Resources within Health Sector Reform promoted by the Pan American Health Organization and other groups, countries of the Region of the Americas have now begun to gather, in a methodical fashion, tangible evidence of the decisive importance of human resources within health sector reform initiatives and particularly of the impact of these initiatives on health personnel. This mutual influence is the main theme of this article, which explores the most disturbing aspects of health sector reform from a human resources perspective, including job instability and conflicting interests of employers and employees. PMID- 11026775 TI - [Payment systems for health care providers in Latin America and OECD countries]. AB - The system used to pay health services providers is one of the most important components of the contractual relationship between persons who receive health services and the individual practitioners and institutions that provide those services. That payment system is also relevant in assessing a health system, including its efficiency and quality. In this article we present a simple analytical framework for various payment systems. We also provide an overview of the payment approaches used in two groups of countries whose experiences we consider representative: 10 nations of the Organization for Economic Cooperation and Development (OECD) and four countries of Latin America. We present a basic model to characterize the different forms of payment based on two dimensions. One of the dimensions is the payment "unit," which is used to measure the amount of health care services provided or promised. The other dimension is the distribution of financial risks between the service provider and the service purchaser. Each payment system has advantages and disadvantages that should be evaluated in relation to the intended objectives. On one extreme of the approaches is fixed remuneration, without any adjustments; it represents the purest prepayment approach. One example of fixed remuneration is capitated payment, in which providers carry all the financial risks coming from the variability in the cost of providing services. On the other extreme is fee-for service payment, where service providers are not at financial risk; the insurer or other financing institution carries all the risk from variable costs. Neither of the extremes appears to be the best choice, and so the issue becomes one of selecting a remuneration system that falls between those extremes. Therefore, it is necessary to choose, on the one hand, the optimal payment unit according to the objectives of the financing entity and, on the other hand, a risk distribution approach that allocates to the service provider the risks coming from greater or less efficiency in delivering services. PMID- 11026776 TI - [Spending and financing in health care: situation and trends]. AB - Being knowledgeable about national health expenditures and sources of financing is essential for decision-making. This awareness also makes it possible to evaluate the equity of allocation and the efficiency of utilization of these resources. Changes in financing have been a substantial component of health sector reform in the Americas. The goal has shifted from merely one of financial sustainability to simultaneously seeking equitable access to quality services. In this article the Pan American Health Organization (PAHO) presents a proposal for analyzing and designing a policy on health financing. The aim of the policy is to identify the mix of financing mechanisms most likely to simultaneously produce financial sustainability, equity, access, and efficiency. The PAHO proposal combines traditional mechanisms for generating resources (public funds from taxes, as well as private health insurance, national health insurance, and user fees) with complementary subsidy mechanisms for vulnerable groups. Health financing strategies ought to explicitly consider the financing both of care for individuals and of health interventions for the general public good, for which public financing is the most equitable and efficient approach. PMID- 11026777 TI - Decentralization of health systems in Latin America. AB - Decentralization is often a major part of health reform policies. However, there have been few attempts to comparatively study the degree of decentralization and the effects of decentralization on equity of allocations to health, so we do not know how best to implement this reform. This article uses an innovative comparative analysis of the "decision space" that was allowed to local municipalities in the health reforms of Bolivia and Chile, two countries that have had several years of experience in implementing decentralization. The studies found that relatively little decision space was allowed to local authorities over key functions of health care systems. The studies also found that central authorities often reduce the decision space in order to direct more resources to health or to restrict local choice over human resources issues. The studies found that more equitable allocations of health funding were achieved through a common equalization fund for the municipalities in Chile and by forcing the assignment to health of a specific percentage of the central government transfers to municipalities in Bolivia. PMID- 11026778 TI - [Quality and health care reform in Latin America and the Caribbean]. AB - Improving the quality of basic health services, together with the search for equity, efficiency, sustainability, and social participation, has been one of the guiding principles of health sector reform initiatives ever since the I Summit of the Americas was held in 1994. This article addresses some basic concepts, examines the status of quality control within health systems and services in Latin America and the Caribbean, and analyzes the most important trends observed in the Region in the establishment of quality assurance programs. Finally, ways of improving and monitoring quality continuously and sustainable are recommended. PMID- 11026779 TI - [Health care reform and social participation]. AB - This article begins by briefly reviewing different forms of citizen participation in the health system. The article then suggests the need for mechanisms for society to control each of the tasks that the health system should perform: defining overall policies, financing, insurance, service provision, and evaluating the effectiveness and efficiency of the system. In general, in the countries of Latin America and the Caribbean, health sector policy formulation continues to be carried out in a centralized manner, with limited involvement by individuals and little adaptation to local realities. The reform processes in the Region are progressing in defining or improving the financing responsibility so as to better reconcile the objectives of equity, efficiency, and freedom of choice. Nevertheless, little has been done to develop instruments for citizens to control the functions of financing and insurance. Appropriate instruments are still lacking for citizens to effectively manage service providers. Among the principal obstacles to citizen participation in the health sector in the Region are: inequality in the distribution of income and other forms of power, an embryonic recognition of the rights of people when they are dealing with public services, limited information concerning those rights and the absence of mechanisms for people to truly exercise them, the weakness with which existing control mechanisms recognize social diversity and incorporate the views of minorities or of the most marginalized groups in society, and weak civic organizations. PMID- 11026780 TI - [Social scenarios in Latin America and the Caribbean]. AB - A wide range of sources have pointed out the magnitude and depth of the social problems that trouble Latin America and the Caribbean and the risks that these problems pose for democracy. Although there are other issues that merit consideration, this article briefly outlines nine key social problems: 1) the increase in poverty, 2) the impact of poverty, 3) unemployment and informal employment, 4) deficiencies in public health, 5) problems in education, 6) the newly poor, 7) the erosion of the family, 8) increasing crime, and 9) the perverse cycle of socioeconomic exclusion. Solving these problems must not be delayed. It is urgent to move to a comprehensive view of development that achieves a different balance between economic and social policies, and that recognizes the indispensable role of these policies in achieving truly sustainable development. At stake are problems that not only concern resources, but also priorities, levels of equity, and the organization of society. Facing up to this poverty and inequity requires an in-depth assessment of these economic policies' social consequences, of the crucial subject of Latin American inequity- the greatest in the world--and of the role of social and public policies. PMID- 11026781 TI - [Towards a new social security in the 21st century: without financing solidarity there will be no universality]. AB - In the countries of Latin America and the Caribbean, social security systems have traditionally been almost exclusively the responsibility of the public sector. These systems have had major shortcomings, such as low coverage rates, unbalanced budgets, inadequate funding, and poor management of resources. In order to solve these problems and face the increased demands associated with demographic and epidemiological transitions, in the 1990s a number of countries began to reform their social security systems. These reforms have been characterized by three fundamental features: a) a search for a closer link between contributions and benefits, in order to better balance income and expenses, b) changes in the public-private composition of the systems that allow a greater private role in the financing and delivery of services, and c) an emphasis on market mechanisms as a way to promote efficiency in applying resources, and leaving to the State a role as a regulator and as a guarantor of basic benefits to groups that, because of their socioeconomic conditions, cannot make certain minimum contributions. This article looks at some of the problems raised by the reforms carried out so far, and the lessons that can be learned from them. The piece also analyzes the relationship between universal coverage and societal unity. In addition, the article suggests that the main challenge with social security reforms is that of moving toward universal systems that significantly expand coverage. In order to achieve that, it is necessary to strengthen the mechanisms of cohesion in financing and to improve efficiency by introducing market instruments that do not negatively affect the unitarian character of the financing. The piece concludes that it is necessary to increase coverage; improve management; be concerned about the design of the public-private makeup; identify the responsibilities of the private sector and of the government in financing, provision, and regulation of social security systems; and introduce and strengthen unity mechanisms in financing. PMID- 11026782 TI - [Rethinking social protection in health in Latin America and the Caribbean]. AB - Despite what is written in the constitutions and other basic document mandates of the countries of the Region, exclusion from social protection in health (SPH) affects an important proportion of the population (at least 20%, which represents, in absolute figures, between 80 and 200 million people). These estimates are obtained through a series of theoretical (social security coverage) and practical indicators that encompass structural indicators (poverty, ethnicity, and geographical barriers) as well as process indicators (non institutional births, compliance with vaccination schedules, and access to basic sanitation). Exclusion levels in a society are affected by the degree of segmentation of the health system. Traditionally, most countries of the Region have had a public, a social security and a private subsystem in health. Lack of attention to the problem has resulted in the formation of a community-based subsystem. The coexistence of many subsystems, along with poor regulation on the part of health authorities, has resulted in high levels of exclusion and inefficient resource allocation within the sector. The organization of social dialogue processes focusing on SPH within the context of health sector reform initiatives in each country is recommended. The process, which should be participatory, should include a full diagnosis of the situation (how many are excluded, who are they, and why, and what mechanisms are the most appropriate for tackling the issue in each country). It should also provide a political and technical feasibility analysis of the most suitable options for each society, and a determination of whether or not conventional subsystems have exhausted their potential. The process should culminate in a program for implementing the specific proposals made in each society, in an effort to maximize SPH. PMID- 11026783 TI - [Essential functions of public health: emerging topic in health care reform]. AB - In the Americas, health sector reforms are facing the challenge of strengthening the steering and leadership role of health authorities. An important part of that role consists of fulfilling the essential public health functions (EPHFs) that are incumbent on all levels of government. For that, it's crucial to improve public health practice, as well as the instruments used to assess the current state of public health practice and the areas where it needs to be strengthened. For that purpose, the Pan American Health Organization has started an initiative called "Public Health in the Americas," with the objective of defining and measuring EPHFs, as a way of improving public health practice and strengthening the leadership provided by health authorities at all levels of government. This article summarizes conceptual and methodological aspects of defining and measuring EPHFs. The article also analyzes the implications that measuring performance on these public health roles, responsibilities, and activities would have for improving public health practice in the Americas. PMID- 11026784 TI - [Regional initiative on health care reform in Latin America and the Caribbean]. AB - Many countries throughout Latin America and the Caribbean are introducing reforms that can profoundly influence how health services are provided and who receives them. Governments in the region identified the need for a network to support health reform by building capacity in analysis and training, both at the Summit of the Americas in 1994 and at the Special Meeting on Health Sector Reform, which was convened in 1995 by an interagency committee of the Pan American Health Organization/World Health Organization, the Inter-American Development Bank, the World Bank, and other multilateral and bilateral agencies. In response, in 1997 the Pan American Health Organization and the United States Agency for the International Development launched the Latin America and Caribbean Regional Health Sector Reform Initiative. The Initiative has approximately US$ 10 million in funding through the year 2002 to support activities in Bolivia, Brazil, the Dominican Republic, Ecuador, El Salvador, Guatemala, Haiti, Honduras, Jamaica, Mexico, Nicaragua, Paraguay, and Peru. Now in its third year of implementation, the Initiative supports regional activities seeking to promote more equitable and effective delivery of basic health services. PMID- 11026785 TI - A roundtable discussion on international cooperation in health reform in the Americas. PMID- 11026786 TI - [Towards a new generation of reforms to improve population health]. PMID- 11026787 TI - [When each person thinks he is less exposed to risks than others, but more receptive to health prevention messages]. AB - Comparative optimism consists of projecting oneself into a situation that is advantageous in relation to others. This is observed in the estimation of future risks, as well as in the evaluation of prevention messages: people who consider themselves less exposed to risks but more receptive than others to prevention messages. Our experimental research, which included a sample of 523 people, examines the effect of prevention messages on comparative optimism regarding future risks such as automobile accidents, domestic accidents or infectious diseases. Three independent variables were tested: the framing of messages, the context of judgement, and the target of comparison. The framing of messages consisted of centering the arguments on losses resulting from the absence of prevention or on the benefits of prevention. The context allowed for the evaluation of the perception of risks with or without a prevention message or reading instructions. People should have indicated if they were more or less likely to be at risk compared to someone close to them (a friend) or to an abstract entity (someone of their generation). Our study shows that the preliminary attention to prevention messages diminishes comparative optimism relative to future risks when the target of comparison is abstract, and the instruction increases the personal implication; it increases the comparative optimism regarding future risks when the comparison is to someone familiar and the messages are framed negatively. The discussion relates to the applications of these results for the conception and evaluation of prevention messages. PMID- 11026788 TI - [Surveillance of laboratory based infections by biological and medical analyses: review of the literature]. AB - Infectious disease surveillance is one of the priorities of public health programmes. This review aims to describe laboratory-based infection surveillance systems. The following surveillance networks are described: the Belgian microbiology network, AIDS surveillance in Philadelphia, EPIBAC, and the Salm-Net network. Laboratory-based surveillance provides the advantage of constituting an easily accessible computerised data bank that allows for studying age-old tendencies and evaluating the effectiveness of certain prevention programmes. The main inconveniences remain with the difficulty of eliminating duplications as well as the lack of clinical information. These difficulties can be surmounted by improving communication systems among the diverse participants in the network with electronic mail. Such surveillance systems will facilitate information exchanges at both the national and international level. PMID- 11026789 TI - [Benchmarks for evaluating health care networks]. AB - The implementation of health care networks is accompanied by a certain number of changes in professional practices. At the heart of the economic, organisational, and political stakes, new questions are posed, especially in terms of evaluation. It is true that no matter what the level of evaluation, it is essential today to rely on the support of existing information systems to develop methodologies and to better describe the produced effects, in economic terms and health outcomes, but also from a more qualitative point of view, in terms of public satisfaction and benefits to clients. Laying down some reference points for evaluating health care networks can only contribute to developing new strategies for showing their effectiveness and efficiency and learning from this experimentation. PMID- 11026790 TI - [Difficulties with the implementation and institutional stakes in the continuing education of private physicians]. AB - The Formation Medical Continue (FMC) is a continuing education programme for private practice doctors (generalists and specialists), and was established in the legislation of April 1996. In analysing the stages of the policy's introduction, an attempt was made to understand the difficulties encountered in its implementation. Using a semi-directive questionnaire, the opinions concerning the policy were collected from the different actors in the process: professionals, social welfare organisations, government agencies and health organisations. The analysis of their perceptions was complemented by an analysis of historical data and a review of the literature. The greatest difficulties with implementation related to the interactions among the stakeholders: the "game" of the medical unions, among themselves and with social security and government agencies; the under-representation of the medical profession, exacerbated by its divisions and the split between generalists and specialists; the indecision of government agencies, leaving the stakeholders waiting; the "game" of the social security funds, which act as if the principle of "mutualisation" of FMC funds can be by-passed. Conflicts of interest regarding the FMC have "crystallised" among the different stakeholders, as well as within the medical corpus. These conflicts relate in particular to the creation of the memorandum agreement and to the definition of the relationship with the pharmaceutical industry. PMID- 11026791 TI - [Interpretations of coordination as an evaluation tool in an experimental gerontologic program]. AB - This article deals with the interpretations of coordination among actors solicited to participate in an experimental programme in the gerontological area of Lunel of the Herault department of France. The article presents the conclusions of an evaluation report which includes two survey phases: a pre-test and a post-test survey implemented at the end of the programme with a parallel survey implemented in a control site, the Pezenas area of the Herault department. The analysis of the initial perceptions of coordination and their evolution relies on the theory of the plurality of worlds (L. Boltanski and L. Thevenot). It aims to show that making a move toward coordination is a complex phenomenon that requires the elaboration of a common language. The material collected during this evaluation allows us to suggest that coordination questions the actors on their professional situation. It questions institutions and their internal functioning. Finally, this type of intervention brings to light the stakes in the examined field. PMID- 11026792 TI - [Use of a self-administered quality of life questionnaire in a university preventive service]. AB - In France, the student population represents close to two million people covered by the medical and social plan of the Services de Sante Inter-Universitaires (Inter-University Health Services), of which one of the objectives is to improve students' quality of life. This is why it seemed interesting to measure this quality of life in order to identify problems and to propose possible improvements. For this, we used a generic self-administered questionnaire, the SF 36, which allowed for detecting the variations of quality of life in relation to health status. Throughout the 1995-1996 school year, 1301 questionnaires were completed by first year students seen in routine visits or scheduled appointments. The questionnaire was well received. The self-administered questionnaire is a sensitive tool that revealed, among healthy students, statistically significant differences in scores between girls and boys (with girls showing lower scores). This difference is not found in the group of students who came for an appointment for a benign pathology. The causes of these variations remain to be explored. PMID- 11026793 TI - [An innovative training experience in public health at the Cheikh Anta Diop University of Dakar]. AB - The Institute for Health and Development (ISED) at the University of Dakar, was created in 1987. Its mission is to fulfill the public health training needs of Senegalese health care professionals. ISED is responsible for a training program entitled "Certificat d'etudes Speciales" (CES), the equivalent of a Masters of Public Health degree. The "CES" is a two-year program comprised of six modules, which last four months each. The ISED training approach stresses trainee responsibility above all. It is based on the trainee's professional tasks and is oriented toward the problems identified by the trainees themselves. In this sense it is highly adaptive and flexible. Each module is implemented in the same way; initially there is a 15 day residential phase for intensive theoretical courses, followed by an application period of 75 days in the trainee's work place, and then a final residential phase of 15 days for writing and presenting a technical report on the work in the field. This model allows trainees to bring together the theoretical and the practical. The trainers directly supervise each of these three phases. The two residential phases are conducted in the ISED training center, located in Mbour, 80 Km south of Dakar. 103 trainees are enrolled in the program in seven different classes. Of these trainees, 93 are health care specialists (85 physicians 7 pharmacists, 1 dentist). Ninety percent of previous graduates have been civil servants who work for the Ministry of Health in Senegal and the remaining 10% were from Burkina Faso and Togo. The training program is considered by all stockholders, beneficiaries, and relevant financial institutions, to be appropriate, beneficial and successful. PMID- 11026794 TI - [Knowledge and communication in public health. Evolution of scientific knowledge during the dramatic event of contaminated blood. 2--Debates on decisions about new options]. PMID- 11026795 TI - [Diffusion and use of the WHO essential drug list in African hospitals]. PMID- 11026796 TI - Medical professionalism: our badge and our pledge. PMID- 11026797 TI - How do our new graduates prefer to learn? AB - BACKGROUND: Learning preference refers to one's choice of specific learning situations or environments over the other. It is one of the factors needed to be considered in planning curriculum and in designing instructional units. OBJECTIVES: The primary objectives of this study were to characterize the learning preference of recent medical graduates of the National University of Singapore (NUS) and to identify any possible gender differences. This study is likely to be first of its kind in this learner population. INSTRUMENT: Rezler's Learning Preference Inventory. METHODS: Rezler's Learning Preference Inventory was administered among twenty-eight 1997 graduates (male 16, female 12) of NUS. The independent variable was the gender and the dependent variables were the scores in each of the learning categories: abstract, concrete, teacher structured, student-structured, interpersonal, and independent. ANALYSIS: Frequency distribution of the learning preferences was counted manually. Independent samples t-test was used to compare two groups of dependent variables. RESULTS: Ninety-two percent female and sixty-nine percent male respondents preferred concrete learning. Only one male respondent and none of the female respondents preferred abstract learning. Among all the respondents, differences between concrete and abstract categories reached statistical, as well as meaningful, significance (p < 0.0001 and mean score difference of 19.9). Differences between student-structured and teacher-structured, and between interpersonal and independent categories did not reach statistical significance (p value of 0.51 and 0.78 respectively). Female respondents generally showed a trend towards greater preference for concrete and teacher-structured learning than their male counterparts. CONCLUSION: The learning preferences of recent graduates of the NUS is characterized by high inclination towards concrete learning. The results can be utilized in designing instructional methods for this group of learners. PMID- 11026798 TI - Throat swab in the chronic tonsillitis: how reliable and valid is it? AB - INTRODUCTION: The diagnostic test of swabbing the surface of the tonsil as a culture specimen for the determination of the organism responsible for the tonsil infection is still in practice, despite controversy. To date there has been no reports of establishing the reliability and validity of this common diagnostic test by appropriate statistical test of Likelihood ratios taking into consideration the specificity and sensitivity. AIMS: To assess the reliability and validity of throat swab in the diagnosis of bacterial microflora in chronic tonsillitis. PATIENTS AND METHODS: A prospective study of 40 patients clinically diagnosed to have chronic tonsillitis undergoing tonsillectomy was undertaken. The reliability of the throat surface swab was then assessed and validated with the reference (gold) standard of tonsil core culture. RESULTS: The likelihood ratio of this diagnostic test being positive (LR +ve) was 0.84 to similar and 1.3 to general organism pathogen. CONCLUSION: Routine culture of the throat by surface swab in the accurate diagnosis of bacterial flora in chronic tonsillitis is neither reliable nor valid. The clinical implications of this investigation which is still very popular is discussed. PMID- 11026799 TI - Tetracycline and nicotinamide for the treatment of bullous pemphigoid: our experience in Singapore. AB - AIM OF STUDY: To study the efficacy of tetracycline (or doxycycline) and nicotinamide in the treatment of less extensive bullous pemphigoid. METHODS: An open trial of 11 patients with bullous pemphigoid. Treatment was initiated with tetracycline 1.5-2 g/day and nicotinamide 1.5-2 g/day and gradually tapered down. Doxycycline was substituted for tetracycline in patients who could not tolerate tetracycline due to gastrointestinal side effects or headache. RESULTS: 6 out of 11 patients achieved complete response (> 90% decrease in lesions) while another 2 had partial response (50-90% decrease in lesions). CONCLUSION: Tetracycline/doxycycline and nicotinamide is a useful alternative treatment for localized bullous pemphigoid, especially in those whose concurrent medical illnesses preclude the use of systemic corticosteroids. PMID- 11026800 TI - Chest impedance: characteristics of local patients. AB - The Pre-hospital Defibrillation Program in Singapore has in some cases demonstrated a lower amplitude of Ventricular Fibrillation (VF) than considered the norm. The Electrode Skin Impedance (ESI) refers to the skin impedance determined between two electrodes placed at specific positions on the body surface. The objective of this prospective study was to measure the ESI of patients at 5 Hz and 2 kHz frequencies, and assess its change with time from the application of electrodes, the difference between the ESI at two different sets of electrode placement positions and correlation with patient factors. Patients who were 25 years or older and not critically ill had their ESI measured with a modified Heart-Save 911 defibrillator, using signal frequencies at 5 Hz and 2 kHz, at 10 seconds, 1 and 2 minutes after electrodes application. Two sets of positions were used; Position 1 where an electrode is placed in the right infra clavicular region and another just lateral to the apex beat on the left and Position 2, which represents the mirror image of Position 1. 36 each of male and female patients were studied. The mean age and weight were 59.9 +/- 13.5 years and 56.8 +/- 24.1 kg respectively. There was no significant correlation between the ESI and patients' body weight or sex. However, there was a significant decrease in the ESI with time from application of electrodes at both Positions (p < 0.05) with the two different frequencies. The ESI was lower when measured at lower frequencies and higher when taken at higher frequencies, but there was no statistically significant difference between the two mirror-image positions used. Thus, with lower frequency, the ECG amplitude of VF recorded on the automated external defibrillator could be enhanced. PMID- 11026801 TI - Clinical pathways--an evaluation of its impact on the quality of care in an acute care general hospital in Singapore. AB - A critical or clinical pathway defines the optimal care process, sequencing and timing of interventions by healthcare professionals for a particular diagnosis or procedure. It is a relatively new clinical process improvement tool that has been gaining popularity across hospitals and various healthcare organisations in many parts of the world. It is now slowly gaining momentum and popularity in Asia and Singapore. Clinical pathways are developed through collaborative efforts of clinicians, case managers, nurses, and other allied healthcare professionals with the aim of improving the quality of patient care, while minimising cost to the patient. Clinical pathways have been shown to reduce unnecessary variation in patient care, reduce delays in discharge through more efficient discharge planning, and improve the cost-effectiveness of clinical services. The approach and objectives of clinical pathways are consistent with those of total quality management (TQM) and continuous clinical quality improvement (CQI), and is essentially the application of these principles at the patient's bedside. However, despite the growing popularity of pathways, their impact on clinical outcomes and their clinical effectiveness remains largely untested and unproven through rigorous clinical trials. This paper begins with an overview of the nature of clinical pathways and the analysis of variances from the pathway, their benefits to the healthcare organisation, their application as a tool for CQI activities in direct relation to patient care, and their effectiveness in a variety of healthcare settings. The paper describes an evaluation of the impact of a clinical pathway on the quality of care for patients admitted for uncomplicated acute myocardial infarction (AMI) through an analysis of variances. The author carried out a one year evaluation of a clinical pathway on uncomplicated AMI in Changi General Hospital (CGH) to determine its effectiveness and impact on a defined set of outcomes. A before and after nonrandomized study of two groups of patients admitted to the Hospital for uncomplicated AMI was done. A total of 169 patients were managed on the clinical pathway compared to 100 patients in the control (historical comparison) group. Outcomes were compared between the two groups of patients. Restriction and matching of study subjects in both groups ensured that the patients selected were comparable in terms of severity of illness. The results showed that the patients on the clinical pathway and the comparison group were similar with respect to demographic variables, prevalence of risk factors and comorbidities. There was a statistically significant reduction in the average length of stay after implementation of the clinical pathway. This was achieved without any adverse effect on short term clinical outcomes such as in-hospital mortality, complication rate and morbidity. There were no significant difference in readmission rates at 6 months after discharge. The paper concludes that clinical pathways, implemented in the context of an acute care general hospital, is able to significantly improve care processes through better collaboration among healthcare professionals and improvements in work systems. PMID- 11026802 TI - Fractures of the medial humeral condyle in adults. AB - BACKGROUND/AIM OF STUDY: Fractures of the medial condyle of humerus are uncommon in adults. The aims of this study were (i) to highlight the rarity of this injury, (ii) to focus on the problems in management of cases which present late, and (iii) to compare the results of surgical excision of medial condyle with those of internal fixation. METHODS: Seven adult medial humeral condyle fractures were seen over a 10-year period. Four cases were surgically fixed within three weeks of the injury; one case refused operation. Two cases which presented late, one with an isolated trochlear fracture, and another Milch type I fracture comminution and compounding, were treated by excision of the condyle and supervised post excision physiotherapy. RESULTS: All six operated cases regained good function. The two patients with excised condyle had no significant instability and had good range of movements. The results were comparable to those managed by open reduction and internal fixation. CONCLUSIONS: Medial condyle fractures presenting early (within 3 weeks), should be managed by accurate open reduction and rigid fixation: non operative management leads to relatively poor results. In late/neglected cases, or those with extensive comminution, open reduction and fixation may lead to stiff and painful elbow, whereas excision of the condylar fragment does not lead to instability. PMID- 11026803 TI - Focal nodular hyperplasia: an unusual liver mass in a paediatric patient. AB - We report an unusual case of focal nodular hyperplasia (FNH) occurring in a 19 month-old female without a typical central fibrous scar. Ultrasound demonstrated a solid, hypoechoic, highly vascular mass situated in the left lobe of the liver. Computed tomography showed a solid mass with no evidence of a central fibrous scar. A hepatic angiogram demonstrated enlarged right and left hepatic arteries supplying the vascular tumour, with early venous drainage into the inferior vena cava; a feature which has not been previously described. Surgical resection was carried out and a solid, nodular tumour measuring 9 x 4 x 4 cm was removed. The diagnosis of FNH was made histologically. The characteristic imaging findings of FNH will be discussed and a review of the literature of FNH in children will be presented. PMID- 11026804 TI - Anaesthesia for caesarean section in a patient with Eisenmenger's syndrome. AB - Eisenmenger's syndrome was originally described in 1897 and redefined by Wood in 1958. This syndrome includes pulmonary hypertension with reversed or bi directional shunt associated with septal defects or a patent ductus arteriosus. A 27-year-old G2 PO with Eisenmenger's syndrome presented to the hospital for management at 17 weeks of pregnancy. She was advised termination of pregnancy but she refused. An elective caesarean section was performed successfully under general anaesthesia uneventfully at 29 weeks due to severe intrauterine growth retardation (IUGR). Patient's postoperative complications like pulmonary thromboembolism, the advantages and disadvantages of anticoagulation are discussed. Pregnancy carries substantial maternal and fetal risk for patients with pulmonary hypertension and Eisenmenger's syndrome. Although pregnancy should be discouraged in women with Eisenmenger's syndrome it can be successful. PMID- 11026805 TI - Brief acute psychosis following hysterectomy in ethnopsychiatric context. AB - The psychiatric morbidity following hysterectomy has received increasing attention. One of the sequelae of hysterectomy has been a brief, acute psychosis with excellent outcome, the etiology and pathomechanism of which is still unclear. Two Chinese patients born of Southeast Asian origin who manifested brief, acute psychosis following hysterectomy are presented. Therapy comprised drug treatment with low dose antipsychotics and benzodiazepines coupled with hypnosis and marital therapy to explore and treat the underlying pathology. Both psychotic states resolved. Follow-up at 12 months revealed stable mental condition in one subject; however, the second patient was lost to follow up. The impact of the womb's removal is explored in the context of the ethnicity of the patients and their sociocultural background. PMID- 11026806 TI - Clinics in diagnostic imaging (51). Multiloculated cerebral abscesses due to paradoxical cardiac emboli. AB - A 19-year-old woman with congenital cyanotic heart disease (Epstein anomaly and ventricular septal defect) had persistent headaches and intermittent fever for five weeks. Physical examination revealed central cyanosis, finger clubbing and fever. The leucocyte count was elevated. Cerebral MR imaging showed the characteristic features of brain abscesses in the right frontal lobe, including multiloculation with adjacent satellite lesions, ring enhancement, T1 hyperintense and T2-hypointense signal areas within the abscess rim, as well as hypointense internal concentric rings on T2-weighted images. The diagnosis of brain abscesses was confirmed by craniotomy and pus drainage. She made a good recovery with a combination of antibiotics and surgery. Follow-up CT scans showed only changes of encephalomalacia at the healed abscess site. The aetiology, clinical features, and the role of CT and MR imaging in the diagnosis and management of brain abscess are discussed. PMID- 11026807 TI - Hippocrates revisited. PMID- 11026809 TI - Physician labor organizations: desperate doctors seek relief through negotiation. PMID- 11026808 TI - Would it have affected the outcome? (loss prevention case). PMID- 11026810 TI - Financial woes increase faculty turnover at medical schools. PMID- 11026811 TI - Loss prevention case of the month. The question: not what but how? PMID- 11026812 TI - Health care on a tightrope: is there a safety net? Part II: the safety net. PMID- 11026813 TI - New Latino immigration to Tennessee: practicing culturally sensitive health care. PMID- 11026814 TI - Newborn screening for congenital adrenal hyperplasia. PMID- 11026815 TI - A case of pneumococcal meningitis. PMID- 11026816 TI - [Allogenic bone marrow transplantation complications. Part II]. AB - Bone marrow transplantation increase the chances of cure of many hematology and also neoplasms cancers. The procedure is however a cause of expected mortality and morbidity. The complications are represented by mucocutaneous, toxicity graft versus host disease, veno-occlusive disease and most importantly injections consequences all this complications needs to be prevented and treated considering the risk associated to the moderling immunosuppression. PMID- 11026817 TI - [Oligohydramnios and fetal malformation association]. AB - L'Oligaminos is sometimes discovered during echographic exploration in obstetrics. Etiology is dominated by renal malformations, obstructive myopathies and polymalformatives syndromes therapeutic interruption of pregnancy needs to be discussed in case of bilateral renal malformation. PMID- 11026818 TI - [Obesity in children]. AB - The aim of this prospective study is to evaluate the frequency of the obesity in the population of 951 children aged between 6 and 12 years old, scholarized in two primary schools situate in Ariana. We found 50 obese children between them (5.25%). The weight and the height of these children were more than 3 standards deviations compared to the normal. 66% of these children have one or two obese parents and 74% were the oldest or the youngest child in the family. The food intake was more important than the normal and it was hyperglucidic rich in sugar and hyperprotidic with troubles of dietetic comportment. PMID- 11026819 TI - [Childhood acute myeloblastic leukemias. Report of 21 cases]. AB - Between 1989 and 1996, 21 cases with acute non lymphoblastic leukemia (11 males and 10 females) were diagnosed in our institution. Median age was 9 years (range, 2-15 years). Leukocyte count was more than 50,109/l in 47% of cases. According to the French-American-British (FAB) criteria, 7 cases were classified M1, 10 cases were classified M2, 1 classified M4Eo and 3 classified M5. All patients were treated with "7 + 3" protocol and complete remission was achieved in 17 cases (80%), 2 cases (10%) failed to respond and 2 (10%) died during induction. Relapse was observed in 15 cases. The 3-year survival rate was 20% and the relapse-free survival rate was 12% confirming the worse prognosis of this leukemia when treated with standard chemotherapy. PMID- 11026820 TI - [Acute psychotic episode in adolescence: predictive elements of mood disorder progression]. AB - OBJECTIVE: Our aim is to try to determine predictors of an evolution to mood disorder during a first psychotic episode in adolescence. METHODS: We proceed with a retrospective study concerning 21 patients having developed a first psychotic episode in adolescence, and admitted in our service within the period from 1991 to 1996, with a follow up of 2 to 7 years. 8 of them, were diagnosed bipolar disorder meeting the criteria of the DSM IV, the other were diagnosed schizophrenic disorder the two groups of patients we have compared. (Fisher test) RESULTS: The predictors of mood disorder evolution seems to be: An acute onset, a good quality of social integration before the onset of troubles and a thymic exaltation, logorrhea. PMID- 11026821 TI - [Pulmonary complications in sickle cell syndromes]. AB - We report various pulmonary complications occurring in 88 patients with sickle cell disease: 44 Hb SS, 29 Hb Sb and 15 Hb SC. Pulmonary infections were observed in 20% of patients, and initiated the disease in 7% of cases. They were common in children younger than 10 years especially in Hb SS and Hb Sb patients. Bacteria were identified in 50% of cases predominantly S. pneumoniae, Mycoplasma pneumoniae, and Haemophilus influenzae. Acute chest syndrome occurred in adolescents and adults and was the consequence of pulmonary infarct or embolism which should be differentiated from bacterial pneumonia. Restrive syndrome was present in 2/3 of patients. It was associated to hypoxemia in 80% of cases. Prophylactic therapy was of paramount importance on long-term penicillin therapy and on closely-spaced immunization against pneumococci and Haemophilus influenzae. Blood exchange are indicated in severe hypoxemia. PMID- 11026822 TI - [Value of magnetic resonance imaging in neuro-lupus: report of 10 cases]. AB - This study shows the contribution of MRI to diagnosis and follow up of the cerebral lesions in neuro-lupus. Ten cases of neuro-lupus have been reported. The affected patients are 7 women and 3 men having an average age of 34 years. For 8 patients, MRI revealed lesions that were not detected using computed tomography. These lesions were high signal zones on T2, scattered with predilection to deep and sub-cortical cerebral white matter. MRI has also shown cortical and sub cortical cerebral atrophy for 3 cases, cortical and deep infarcts for 2 cases and both atrophy and infarcts in one case. The MRI diagnosis was normal for the two remaining patients. MRI contributes to the diagnosis confirmation and localization of the cerebral lesions and to the follow-up. In spite of MRI sensitivity, the detection of these lesions remains non specific to neuro-lupus. New methods of exploration as the spectroscopy by magnetic resonance and the cerebral scintigraphy with Technetium 99m could be very helpful in diagnosis of cases for which MRI has shown limitations. PMID- 11026823 TI - [Age and experience of puberty in young Tunisian girls]. AB - The puberty is an important period in woman life. We have tried to determine the age of the puberty, the chronology of its events and the real life of this period of preparation to reproductive life of tunisian girl student by an inquiry. The middle age of puberty is 13 years. It depends of sociological, economic and environmental factors. Knowledges about reproductive life are still insufficient for this age group bracket of high risks of non desired pregnancies and sexually transmittable diseases. PMID- 11026824 TI - [Role of lysosomes in the intracellular mineral elements: the case of lanthane]. AB - The subcellular localization of lanthane was studied using two microanalytic methods. 1.) The election microprobe method showed that lanthane is concentrated in the lysosomes of the hepatocytes and the spleen associated with phosphorus. 2.) The ion microanalysis showed that lanthane is selectively precipitated in the lysosomes of the enterocytes optical part. This mechanism of selective precipitation initially prevents the diffusion of this toxic element in the organism and then permits its elimination digestively after desquamation of enterocytes in apoptosis in the intestinal lumen. PMID- 11026825 TI - [Cystic dystrophy in an aberrant pancreas in the duodenal wall causing a megaduodenum]. AB - One case of cystic dystrophy in an aberrant pancreas of the duodenal wall with a megaduodenum is reported. It is a rare affection which is difficult diagnostic, helped by echoendoscopy. Difficult therapeutic dilemma is raised by cystic dystrophy in heterotopic pancreas: duodenopancreatectomy or limited local excision? PMID- 11026826 TI - [Buschke-Loewenstein giant condyloma in the perianal region with inguinal invasion: a case report]. AB - The giant condyloma acuminatum of the perianal region is a tumor characterised by its large size with the propensity to infiltrate into deeper tissues, contrasting with a microscopically benign pattern. The evolution after malignant transformation of condyloma and lymph node invasion is rare. However it is exceptionally observed a lymph node invasion of microscopically benign condyloma acuminatum. The authors report a case of microscopically benign giant condyloma acuminatum of the perianal region associated with inguinal invasion, discovered in a 47 year-old man. Treatment consists in extensive surgery of the tumor and inguinal nodes, followed by a radiation therapy. PMID- 11026827 TI - [Chronic septic granulomatosis in a young girl manifesting as invasive pulmonary aspergillosis and flaccid paraplegia]. AB - The invasive Pulmonary Aspergillosis (IPA) is the principal cause of death in patients with chronic granulomatosis disease (CGD). It can happen before age of one and can reveal the (CGD). Usually, the transmission of GSD is linked to x in 65% of cases. We report, the case of a girl aged 3 years issued from consanguine marriage referred to hospital for hemoptysia. We note that at 40 days she presents an suppurate adenitis. She is hypotrophic at--3DS without respiratory problem, she have a splenomegaly. The radiography chest note an alveolar and interstitial opacity bilateral, apical right and basal left. There is a biological inflammatory syndrome. During the evolution, the child presents a paraplegia secondary to compression myelitis. A D5 laminectomy in urgency was done. The vertebral Biopsia, the serology and the culture isolate fumigatus aspergillus the GCD is evoqued confirmed by the NBT test who was negative. IN CONCLUSION: --We have to look for GCD in front of pulmonary aspergillus.--The vertebra lesion is secondary to pulmonary lesion.--Our observation is noticeable by the recessive autosomal transmission who was seen in 35% of cases, and by the neurologic complication rarely reported. PMID- 11026828 TI - [Foreign bodies in the hand]. AB - The strange body at the hand level is frequent consultation pattern. Its extraction seems easy, however it's not. The authors reported one observation of the hand which is penetrated by a ig strange body. It's ablation is performed in a surgical room under anaesthesia, respecting the tension lines at the hand level and avoiding the nerves ramification of the cubital nerve that was hung by the harpoon. PMID- 11026829 TI - [Day hospital: why, for whom, which structure?]. PMID- 11026830 TI - [Diabetes and the day hospital]. AB - The day hospital is a relatively new way of hospitalization in Tunisia, the first experience beginning in 1985 to the National Institute of Nutrition. This hospitalization avoid the drawbacks of classic hospitalization (dependency, discomfort, separation) and boredom and present a lot of advantages of social command, humanitarian, psychological, medical and economical the cost of hospitalization is clearly reduced). This day hospitalization is beneficial in several pathologies notably the illness nutrition and metabolic diseases (diabetes, obesity, dyslipoproteinemia, hyperuricemia), where the patients continue to have a good physical activity and where the education medico sanitary and dietary hygieno occupies a position of choice. PMID- 11026831 TI - [Day hospital in medical oncology]. AB - Day hospital or ambulatory or outpatient therapy represent an important part of the activity in oncology in addition to the inpatient approach. PATIENTS AND METHODS: We analyse the data of the medical oncology day hospital from december 1995 to february 1996 in our institution. RESULTS: We collect 2661 consultants in medical oncology and 816 (31%) have been oriented to the day hospital for chemotherapy mainly adjuvant for breast cancer (68%), colo-rectal cancer (18%) or lymphoma (5%). All treatment are administered under antiemetics of setron types for 40% of the patients. We perform also transfusion of red cells or more rarely platelets. Day hospital cover also diagnostic procedures as bone marrow punctures or biopsies or therapeutic gests as pleural or lumbar punctures. CONCLUSION: Ambulatory chemotherapy represent an important component in medical oncology needing an adequate organisation and team synchronisation. PMID- 11026832 TI - [Ambulatory surgery in the hospitals of Tunis, future perspectives]. PMID- 11026833 TI - [Ambulatory anesthesia: which optimal organization for optimal safety?]. AB - The ambulatory anesthesia is now well developed because of the improvement of the intraoperative and the postoperative anesthetic management. The environmental conditions are as well important as surgery and anesthesia management. The ambulatory surgery may be practiced in different type of structures. The conception of an ambulatory center must reflect the medical need and must take care of the economical and social conditions. The aim of this review article is to determine the benefit and the disadvantage of the types of ambulatory surgery organization. PMID- 11026835 TI - [Experience of ambulatory anesthesia in Charles Nicolle hospital]. AB - The authors report the experience of ambulatory surgery at Charles Nicoll's hospital on ophthalmology and ORL surgery in which 90% and 60% of acts can be realized on ambulatory. The organisation by means of structures and hospital workers makes an important gain. The anesthetic consultation, very important in this organisation, enable to avoid to rescue a patient at the last time. PMID- 11026834 TI - [Ambulatory anesthesia in pediatric surgery]. AB - Child is an ideal patient for day care surgery. So more than 60% of paediatric surgery could benefit by ambulatory surgery. Preoperative visit may select patients for ambulatory surgery. Medical exam may lead to choose pre operative screening. The ideal ambulatory anesthesia is locoregional technic or inhalatory one. Tracheal intubation don't contre indicate ambulatory surgery. Recovery of mental abilities following general anesthesia has not the same significance as in adult. Many studies confirm the safety of paediatric outpatients anesthesia. PMID- 11026836 TI - [Plastic surgery in ambulatory practice]. AB - We have analysed 283 patient studies operated of ambulatory during the last two years. These patients represent only 12% of the patient group. It's especially a matter of patients operated urgently. The local anaesthesia in the regional local was practised in terms of 90% of cases. The complications where exceptional. We wanted to insist on the necessary guidelines and pitfalls to forget during the practice of that surgery and we wanted to focus on its advantages. PMID- 11026837 TI - [Ambulatory urological surgery]. AB - Ambulatory surgery offers to patients to be treated on an outpatient basis. Strict criterious, social, surgical and medical must be associated to authorize this type of hospitalisation. From January 1998 to December 1999, 1882 patients (46% of our operative activity) were treated on an outpatient basis, to undergo an endoscopic surgery (66%), conventional surgery (20%) or other (14%). 9 patients remain a supplementary night because of post-operative fever in 5 patients and an incomplete recovery in 4 patients. No patient was rehospitalized for complication. In this series, ambulatory surgery has proved its advantages, without loss of patients' security, if selection criterious were respected. PMID- 11026838 TI - [Ambulatory surgery of hand pathology. Experience of the hand surgery service of the Kassab Institute]. PMID- 11026839 TI - [Ambulatory surgery in gynecology]. PMID- 11026840 TI - [Patients' opinions about day cataract surgery in a general hospital]. AB - OBJECTIVE: To evaluate the patient satisfaction as a performance indicator of quality of health care delivery in outpatient cataract surgery. PATIENTS AND METHODS: We performed a prospective study using a questionnaire containing 14 items (preoperative work-up, administrative formalities, local anesthesia..) evaluating all the phases of ambulatory surgery for cataract. RESULTS: We collect 75 responding patients and we observed 2/3 of relative satisfaction about outpatient cataract surgery. Negative opinion was in relation with the prolonged time required for preoperative work-up which is done outside the outpatient surgery structure. CONCLUSION: Advances in techniques of locoregional anesthesia and surgery permit today the frequent use of ambulatory treatment in cataract surgery. This method seems to be well accepted by patients. However the prolonged time used to do the preoperative work-up require in the future a centralisation for this procedure. PMID- 11026841 TI - [Medical research in Tunisia. Organization and legislative aspects]. PMID- 11026842 TI - [Emergency care organization in Tunisia and international models]. PMID- 11026843 TI - [Non-tumor ano-rectal pathology. Tunisian report]. PMID- 11026844 TI - Orthopod or orthogod? A prospective questionnaire study of post-operative patients' impressions of orthopaedic surgeons. AB - A questionnaire that challenged various stereotypes within the medical profession that are attached to orthopaedic surgeons was distributed to two groups of post operative trauma in-patients: those with osteoporotic fractures and a much younger and active group. The results are presented here in a format that allows the reader to draw his/her own conclusions as to how 'orthopods' are perceived by their patients. PMID- 11026845 TI - Orthopaedic specialist registrar selection: the advantages of a candidate ranking procedure at interview. AB - For five years an orthopaedic specialist registrar appointments committee has been using a points system to rank candidates at interview. By so doing committee members have been able to devote the discussion period, after all candidates have been interviewed, to considering the merits of those candidates who have achieved the greatest degree of support. This system maximises the efficient use of committee discussion time without restricting the freedom of the committee to make the most appropriate appointments. PMID- 11026846 TI - FDS Study Day care of the medically and dentally compromised child. Thursday 16 March 2000. PMID- 11026847 TI - The black box of the ITU. PMID- 11026848 TI - Intensive care and the surgical trainee. PMID- 11026849 TI - Focusing the mind. PMID- 11026850 TI - The changing face of surgical research. PMID- 11026851 TI - Current attitudes to surgical needlestick injuries. PMID- 11026852 TI - Workshop on medical X-ray imaging: potential impact of new EC directive. Report on a meeting held in Malmo, Sweden, on 13-15 June 1999. PMID- 11026853 TI - Proton beam radiotherapy. PMID- 11026854 TI - MRI of the breast. AB - MRI of the breast is rapidly evolving as a sensitive technique for the detection and staging of breast cancer. It has considerable benefits but is time consuming and expensive. The benefits and limitations of this technique are presented. PMID- 11026855 TI - A comparison of bolus chasing and static digital subtraction arteriography in peripheral vascular disease. AB - The purpose of this study was to compare the techniques of bolus chasing angiography (BCA) and digital subtraction angiography (DSA). 75 patients with symptomatic atherosclerotic peripheral vascular disease were randomly assigned to have their lower limbs examined by BCA or DSA. Dose-area product (DAP), time of examination and dose of contrast medium were measured. Staff doses were measured with personal electronic dosemeters. Image quality was assessed from the laser printed images. DSA produced better images of vessels below the inguinal ligament, particularly the crural vessels (1/76 vs 17/74 non-diagnostic examinations, p < 0.00001) but at a higher DAP (median 53.8 Gy cm2 vs 18.9 Gy cm2, p < 0.01). Contrast medium dose was higher with BCA (29.8 gI2 vs 25.3 gI2, p < 0.01). Staff doses per unit patient dose were 2.3-3.3 times higher with BCA than DSA. Because of the poor long-term prognosis of patients with peripheral vascular disease, the improved image quality obtained by DSA justifies the increased radiation dose. PMID- 11026856 TI - Comparison of cardiac stroke volume measurement determined using stereological analysis of breath-hold cine MRI and phase contrast velocity mapping. AB - Cine MRI of the heart using segmented k-space pulse sequences permits multiphase images to be acquired in a single breath-hold. Whilst image quality is improved compared with conventional (non-segmented) cine imaging, subsequent analysis can be relatively time consuming. In this study, multiple slice breath-hold cine imaging of the heart was performed in 11 normal volunteers. Left and right ventricular stroke volume (SV) was estimated from the images by application of the Cavalieri method of modern design stereology in combination with point counting. The measured SVs were compared with those obtained by cine phase contrast velocity mapping in the ascending aorta and main pulmonary artery. Excellent agreement was found between the SVs determined by the two techniques, with mean differences (+/- one SD) of 0.41 +/- 3.00 ml and 0.41 +/- 4.78 ml for left and right ventricles, respectively. Comparison of left and right ventricular SV using stereology yielded a mean difference of 0.84 +/- 5.70 ml. Breath-hold data acquisition together with stereological analysis is demonstrated to be an accurate and unbiased technique for the rapid assessment of cardiac function. PMID- 11026858 TI - Value of spiral CT in patients with cricothyroid approximation. AB - Cricothyroid approximation raises the vocal pitch by simulating contraction of the cricothyroid muscle with sutures. The aim of this study was to determine the role of spiral CT in patients scheduled for cricothyroid approximation. 29 transsexual patients were examined with spiral CT prior to and after laryngoplastic surgery. CT findings were correlated with phonographic findings in all patients. The average reduction of the cricothyroid distance was 6 mm (range 2-10 mm). Vocal pitch elevation was greatest in patients with the largest reduction of the cricothyroid distance. CT accurately determines the cricothyroid distance prior to and after surgery and is an ideal method for follow-up, especially in post-operative reversion towards a lower pitch. In addition, CT provides important data in determining the most appropriate extent and site of intracordal intervention to achieve a desired pitch elevation. PMID- 11026859 TI - A novel quality assurance method in a university teaching paediatric radiology department. AB - Primary diagnostic equipment in a paediatric radiology department must perform at optimal levels at all times. The Children's Hospital Radiology Department in Winnipeg, Canada, has developed an impartial means of reporting radiographic image quality. The main objectives of this study programme were two-fold. First, to monitor diagnostic X-ray equipment performance, and second, to improve the resultant image quality as a means of implementing the fundamental concepts of continuous quality improvement. Reading radiologists completed a quality assurance (QA) card when they identified a radiographic image quality problem. The cards were subsequently collected by the clinical instructor who then informed, in confidence, the radiographers of the written comments or concerns. QA cards have been conspicuously installed in the paediatric radiology reading room since the middle of 1993. Since its inception, equipment malfunction has been monitored and indicators for improving image quality developed. This component of the QA programme has shown itself to be a successful means of communicating with radiographers in maintaining superior image quality. PMID- 11026857 TI - Percutaneous ethanol injection of the supplying artery to hepatocellular carcinoma that is not amenable to conventional treatment. AB - The purpose of this study was to evaluate the clinical usefulness of ultrasound guided percutaneous ethanol injection of the supplying artery (PEISA) to the tumour in the palliative management of hepatocellular carcinoma (HCC) that is not amenable to conventional treatments. A total of 23 cases of HCC, measuring from 3.1 cm to larger than 15 cm (median 5.4 cm) in 17 cirrhotic patients, were treated by PEISA. PEISA was used to control rapid growth of the tumour in seven patients and to reduce abdominal discomfort caused by rapid expansion of the tumour in 10 patients. Tumours with arterial Doppler signals persisting after PEISA underwent repeated treatment. The follow-up period ranged from 2-48 months. PEISA was achieved in 69 out of 76 attempts (90.8%). The amount of ethanol injected on each occasion ranged from 2.5-33 ml. Follow-up colour Doppler scanning showed complete elimination of tumour Doppler signals in 22 out of 23 lesions (95.7%). Following treatment, one tumour disappeared, 13 tumours shrank and nine tumours were unchanged in size. All patients with abdominal discomfort had relief after treatment. The common complications of PEISA were local pain and fever. In conclusion, PEISA is effective at treating painful HCC unsuitable for conventional treatment. PMID- 11026860 TI - Radiation doses to patients undergoing scoliosis radiography. AB - In this study we computed the radiation doses associated with scoliosis radiography and investigated how these radiation doses are influenced by the weight of the patient. We recorded the radiographic technique factors of 61 consecutive patients (46 females and 15 males) undergoing scoliosis radiography. A wedge-shaped aluminium filter attenuated the X-ray beam in the "chest region" relative to the "abdomen region". X-ray tube air kerma output factors (microGy mAs-1) and half value layers (HVLs) were determined experimentally for the "chest region" and "abdomen region". The energy imparted to each patient was computed from the air kerma area product, X-ray beam HVL and measured patient thickness. Values of patient effective dose were obtained using effective dose-to-energy conversion factors for specified radiographic projections, taking into account each patient's weight. The median patient age was 17 years, and the median patient weight was 53 kg. Entrance skin air kerma values in the "chest region" were approximately a factor of four lower than those in the "abdomen region". The air kerma values increased by a factor of two when the patient weight increased from 30 kg to 70 kg. Approximately 80% of the total energy imparted to a patient undergoing a scoliosis examination was in the "abdomen region", with the remaining 20% imparted to the "chest region". Energy imparted increased with patient weight, and was approximately 3 mJ for a 30 kg patient and approximately 8 mJ for a 70 kg adult patient. Effective doses showed little correlation with patient weight, with an average-sized patient (50 kg) receiving an effective dose of approximately 140 microSv. Patients undergoing scoliosis radiography receive effective doses that are low in comparison with other types of radiographic examination. PMID- 11026861 TI - Some limitations in the practical delivery of intensity modulated radiation therapy. AB - The resolution characteristics of intensity modulated beam (IMB) profiles produced by milled compensators and by multileaf collimators (MLCs) are independently investigated with respect to the primary fluence. It is shown that both methods have different characteristics in the longitudinal and lateral direction and, as a consequence, the resolutions of the longitudinal and lateral delivered IMB profiles differ. For both methods, the restrictions are identified. For compensators, the maximum slopes in the machining process, which should not be exceeded, are quantified. For MLCs, emphasis is given to the direction perpendicular to leaf movement. A number of test modulations were created and the effect of different size MLCs on the intensity profile revealed that unacceptable errors can be introduced if the profiles are heavily modulated. The production of intensity modulated radiation therapy (IMRT) beams by both machined compensators or by MLCs is limited by physical constraints. Having identified these constraints, some steps should now be taken to accommodate them either in the objective function for the calculation of the beam profiles or in the delivery system. PMID- 11026862 TI - Vaginal vault brachytherapy: the effect of varying bladder volumes on normal tissue dosimetry. AB - This study was designed to assess the impact of bladder volume on dosimetry to critical normal structures in vaginal vault brachytherapy using a single line source vaginal applicator. 30 consecutive patients undergoing vaginal vault brachytherapy were studied by CT scanning with the applicator in situ and the bladder empty and then with the bladder containing either 35 ml of water (10 patients), 70 ml of water (10 patients) or 100 ml of water (10 patients). The scans were then analysed with isodose distributions overlayed to determine changes in dosimetry. No effect on bladder dose was seen with increasing volume compared with the empty bladder; however, there was a reduction in amount of small bowel within the high dose treatment region as bladder volume increased. With 100 ml bladder volume, the reduction reached 57.5% compared with the empty bladder. We conclude that vaginal vault brachytherapy should be undertaken with a bladder volume of at least 100 ml, which will considerably reduce the amount of small bowel in the irradiation volume with no increase in bladder dose. PMID- 11026863 TI - Accurate measurement of exposure rate from a 60Co teletherapy source: deviations from the inverse-square law. AB - A cylindrical gamma-ray 60Co source of activity alpha is predicted to produce an exposure rate X at a distance d in vacuum, given by X = gamma(T)(alpha/d2), where gamma(T) is the specific gamma-ray constant. It has been documented that this formula may be used to approximate X with an accuracy of 1% from a source of length l, provided that d/l > or = 5. It is shown that the formula is accurate to 0.1% under these conditions, provided that the distance is measured from the centre of the source. When absorption in the source and scattering in the collimator are considered, the position of the origin d = 0 can shift by a distance of the order of centimetres. Absorption in air between the source and the ionization chamber adds an exponential factor to the formula. It is shown that even when these modifications are included the discrepancy in the results, although generally less than 1%, is still large compared with the measurement errors. Some suggestions are made for the origin of this discrepancy. PMID- 11026865 TI - Radiography for midfacial trauma: is a single OM 15 degrees radiograph as sensitive as OM 15 degrees and OM 30 degrees combined? AB - Two previous studies have suggested that a single occipitomental (OM) 15 degrees radiograph may be sufficient for screening for midfacial fractures in the Accident and Emergency (A&E) department, but the study had insufficient power to detect whether some fractures might be missed with only one projection. We undertook to determine whether there is a decreased sensitivity for detecting fractures if only a single OM 15 degrees radiograph is used rather than OM 15 degrees + 30 degrees. 44 cases of major and minor midfacial fractures interspersed randomly with 49 cases of facial injury without fracture were shown to nine A&E clinicians. Initially they were shown only the OM 15 degrees radiograph, then the OM 30 degrees radiograph alone, and then the two radiographs together. They were asked each time to indicate the presence or absence of fracture. The sensitivity for the OM 15 degrees projection alone was 89.4%, and when both radiographs were used it was 90.9% (difference not significant). Given the positive yield of facial radiographs is only 6%, any minimal increase in sensitivity from using two projections is likely to be outweighed by the beneficial reduction in X-ray exposure and costs from using only a single projection. The policy of using a single OM 15 degrees radiograph as a screening tool for midfacial fractures in A&E is a safe one. PMID- 11026864 TI - Gadolinium-DTPA as X-ray contrast medium in clinical studies. AB - The aim of this study was to investigate the contrast effects of gadolinium (Gd) in patients undergoing digital subtraction angiography (DSA), intravenous urography (IVU) and CT. 15 patients attending for coeliac axis DSA (n = 5), abdominal CT (n = 5) and IVU (n = 5) were injected with 0.3 mmol kg-1 Gd-DTPA, the maximum approved dose. For DSA and IVU, images were categorized as being of diagnostic or non-diagnostic quality. For CT, enhancement was measured in Hounsfield Units (HU). On CT, enhancement with Gd was reproducible in all cases; average peak aortic enhancement was 75 HU but duration was short. On IVU, four of five studies yielded positive pyelograms but all nephrograms were relatively poor. On DSA, all five patients had diagnostic arteriograms and four of five indirect portograms were of diagnostic quality. In all 15 cases, enhancement was weaker than that achieved with routine dosage of iodinated agents. In conclusion, Gd chelates may be clinically useful in X-ray studies under certain circumstances on patients with contraindications to iodinated agents. Higher doses than currently approved would be potentially useful. PMID- 11026866 TI - A filter for breast imaging on a radiotherapy X-ray simulator. AB - Simulator radiographs taken as a record of breast radiotherapy planning often show ill defined breast tissue margins because exposure parameters are set to optimize visualization of the chest wall rather than the bulk of the breast. This creates difficulties when using simulator images as reference images in verification by comparing with either portal film or images from an electronic portal imaging device. Our aim was to improve breast images taken at simulation without changing exposure parameters that have been optimized for visualization of the chest wall. This has been achieved via an external filter to be used when taking radiographs with the treatment simulator. The filter is made of stainless steel coated with tin and is shaped to maintain acceptable imaging of the chest wall by covering only the section of field anterior to the chest wall. Radiographs of breast simulations using the filter have been accepted as satisfactory by both clinicians and radiographers. The filter is now in routine clinical use for breast and chest wall treatment simulation. PMID- 11026867 TI - Dose equivalents of tumour repopulation during radiotherapy: the potential for confusion. AB - When employing linear quadratic equations to calculate compensation for changes in overall treatment time, a potential confusion exists regarding use of the parameter commonly described as the dose equivalent of tumour repopulation. The more correct term for this factor is the biologically effective dose equivalent of tumour repopulation. The distinction between the two concepts is discussed and the potential errors arising from their confusion are illustrated by means of an example. PMID- 11026868 TI - Intrafracture fluid: a new diagnostic sign of insufficiency fractures of the sacrum and ilium. AB - MRI is generally regarded as being sensitive but non-specific for the detection of insufficiency fractures affecting the sacrum and pelvic ring. The finding of intrafracture fluid is described in two elderly patients with insufficiency fractures. This MR feature is believed to be a new diagnostic sign of these fractures affecting the sacrum and ilium. PMID- 11026869 TI - Osteoma of the thyroid cartilage--an unusual cause of difficult intubation. AB - We describe a case of histologically proven osteoma of the thyroid cartilage that presented because of difficulty in intubation prior to coronary bypass surgery. To our knowledge, this is the first documented case in the English literature. PMID- 11026870 TI - Metastatic paraganglioma causing spinal cord compression. AB - We describe two cases of metastatic retroperitoneal paraganglioma associated with extradural spinal cord compression. Both occurred in young men; one being metastatic at presentation, the other becoming metastatic 19 years after attempted surgical resection. Despite a long, relatively stable natural history after diagnosis (10 and 19 years, respectively) both had an acceleration of their disease once extradural disease was detected. These cases illustrate the potentially aggressive nature of this disease, the need for long-term follow-up and the effectiveness of a variety of therapies for palliation, and also raise the possibility of "prophylactic" treatment to prevent spinal cord compression. PMID- 11026871 TI - Conservative surgery and radiation therapy for early stage breast cancer after previous mantle radiation for Hodgkin's disease. AB - There is an increased incidence of breast cancer in female patients who have previously undergone mantle radiation for Hodgkin's disease. Lumpectomy followed by breast irradiation is generally considered to be contraindicated in such patients owing to the high cumulative radiation dose to the breast. Mastectomy is therefore recommended as the preferred treatment option in these women. We report two cases of breast cancer occurring in women previously treated with mantle radiation for Hodgkin's disease. Both women declined mastectomy and requested breast-conserving treatment. PMID- 11026872 TI - MRI of retroperitoneal collections: a comparison with CT. AB - Retroperitoneal collections in the absence of pancreatitis are rare. Imaging plays an important role in determining the aetiology of these collections and in allowing percutaneous drainage to be performed safely. A review of the imaging characteristics on both CT and MRI is presented, highlighting the advantages and disadvantages of these two modalities and their complimentary roles. PMID- 11026873 TI - A large abdominal mass in a young girl. PMID- 11026874 TI - Preventing mother to child transmission of HIV: the role of caesarean section. PMID- 11026875 TI - The future of chlamydia screening. PMID- 11026876 TI - The significance of oral health in HIV disease. PMID- 11026877 TI - Overview of study design in clinical epidemiology. PMID- 11026879 TI - A longitudinal study of HPV detection and cervical pathology in HIV infected women. AB - OBJECTIVE: To monitor the presence and persistence of high risk (HR) human papillomavirus (HPV) in cervical brushings from HIV infected women. METHODS: Prospective observational cohort study of HIV infected women. Women were enrolled from the cohort of 164 HIV infected women who attend the colposcopy clinic at the Edinburgh Regional Infectious Diseases Unit. A single cervical brush scrape was obtained from 39 women and two or more samples from 63 women who attended regularly at approximately 6 monthly intervals. HPV typing was carried out using a commercial hybrid capture assay (HCA). Details of antiretroviral therapy, cytological assessment, and histological evaluation were made available and the interrelation with HR-HPV detection analysed. RESULTS: Abnormal cervical cytology, particularly of low grade, was common in these HIV infected women. HR HPV types were detected in 25% of the women with normal cytology, while over 80% of those with abnormal cytology of any grade were HR-HPV positive. Persistent HR HPV, as defined by two or more consecutive HPV positive results, was common and found in 27/63 women from whom multiple samples were obtained. HR-HPV was detected at high levels whether or not patients were receiving antiretroviral therapy. Profound immunosuppression was not necessarily associated with progression of cervical disease and no cases of invasive cervical disease were seen. CONCLUSION: While mild dyskaryosis (low grade squamous intraepithelial lesion (LSIL)) and persistence of HR-HPV are common in HIV infected women in Edinburgh, regular cytological and colposcopic evaluation with appropriate intervention and treatment appears to limit the progression of cervical disease. PMID- 11026878 TI - Trichomonas vaginalis epidemiology: parameterising and analysing a model of treatment interventions. AB - BACKGROUND: Trichomonas vaginalis, which affects at least 170 million individuals globally, may increase the risk of transmission of HIV and predispose pregnant women to premature rupture of membranes and early labour. OBJECTIVE: To more clearly define the epidemiology of trichomoniasis and to develop a mathematical model of disease transmission dynamics in order to explore various treatment strategies. DESIGN: A deterministic model of trichomoniasis was constructed. Parameter values were set to fit the model to known endemic prevalence levels of Trichomonas vaginalis. Two treatment interventions ("screening" and "syndromic management") were simulated. RESULTS: The age specific prevalence of the disease was seen to differ from other STDs in a number of studies. Parameter fitting indicates that the average duration of infection in women is at least 3-5 years and approximately 4 months for men. "Syndromic management" (that is, treating only those with symptoms of disease) had minimal effect upon the endemic prevalence of disease even at high levels of coverage. "Screening" (that is, identification of individuals with both symptomatic and asymptomatic infection) was shown to be the most efficient method of control, but was sensitive to the screening interval. CONCLUSIONS: The control of trichomoniasis seems to have been a success in developed countries because of the regular access to health care, whereas it has remained endemic in many developing countries where control may only be possible by regular screening and treatment. However, without a large investment in services, success in controlling trichomoniasis is likely to be transitory. PMID- 11026880 TI - Comparison of risk factors for four sexually transmitted infections: results from a study of attenders at three genitourinary medicine clinics in England. AB - OBJECTIVE: To compare the risk factors for four common sexually transmitted infections (STIs) in attenders at three large urban genitourinary medicine (GUM) clinics in England. METHODS: Clinical, demographic, and behavioural data on attenders at two clinics in London and one in Sheffield were collected. Risk factors associated with first episodes of genital warts and genital herpes simplex virus (HSV), and uncomplicated gonorrhoea and chlamydia were investigated using the presence of each of these STIs as the outcome variable in separate multiple logistic regression analyses. RESULTS: Using data on the first attendance of the 18,238 patients attending the clinics in 1996, the risk of a gonorrhoea or chlamydia diagnosis was strongly associated with teenagers compared with those aged over 34, with black Caribbeans and black Africans compared with whites, and increased with the number of sexual partners. The risk of genital warts or HSV diagnosis was lowest in black Caribbeans and black Africans compared with whites and was not associated with the number of sexual partners. While genital warts were associated with younger age, odds ratios were much lower compared with those for the bacterial infections. Genital HSV diagnoses were not associated with age. CONCLUSIONS: This study of GUM clinic attenders suggests a reduction in the incidence of bacterial STIs may be achievable through targeted sexual health promotion focusing particularly on black ethnic minorities, teenagers, and those with multiple sexual partnerships. Viral STIs were less clearly associated with population subgroups and a broader population based approach to sexual health promotion may be more effective in controlling these infections. PMID- 11026881 TI - Geomapping of chlamydia and gonorrhoea in Birmingham. AB - OBJECTIVE: To investigate if the core population hypothesis is applicable to patients with genital chlamydia infections. DESIGN: Retrospective cross sectional study. SETTING: Two genitourinary medicine (GUM) clinics in the city of Birmingham and eight adjacent clinics. SUBJECTS: All patients with chlamydia (n = 665) or gonorrhoea (n = 584) attending between 1 October 1995 and 30 September 1996 with a postcode within the Birmingham health district. Controls were 727 patients seen in the same period with no infection. METHODS: Postcodes were used to calculate population prevalence rates per 100,000 aged 15-65 in the 39 wards of the city and to estimate the socioeconomic status using the Super Profile (SP). Ethnic specific rates were also calculated. Data were obtained on gonorrhoea and chlamydia isolation from all the major laboratories of the city over the same time period. RESULTS: GUM clinic attenders accounted for 67.6% and 82.5% of all chlamydia and gonorrhoea isolates reported by the laboratories and that were available for our epidemiological analysis. Both infections were more common in men and in black ethnic groups. However, patients with gonorrhoea only infection were more likely to be of black ethnicity than those with chlamydia only infection (p = 0.0001) and to have different SP distribution (p = 0.0001). On logistic regression age < 20 years, male sex, black ethnicity, and living in neighbourhoods with SP J ("have nots") were predictive of both infections compared with controls. Overall chlamydia and gonorrhoea prevalence rates were 129 and 98.4 per 10(5) respectively. Corresponding rates for whites was 64.7 and 37.2 and for black ethnic groups 1105 and 1183 per 10(5) of each ethnic group. Eight adjacent wards accounted for 41% of the chlamydia and 66.5% of the gonorrhoea. CONCLUSION: In a large urban setting patients attending GUM clinics with chlamydia belong to core population groups with similar, but not identical, sociodemographic characteristics to patients with gonorrhoea infection. PMID- 11026882 TI - Prevalence of Chlamydia trachomatis in young men in north west London. AB - BACKGROUND: Chlamydia trachomatis is the most common, treatable, bacterial sexually transmitted infection in England and Wales. Among men, chlamydial infection is an important cause of non-gonococcal urethritis, epididymitis, and proctitis. The case for wider screening among women has been accepted by an expert advisory group. In the absence of estimates of the prevalence of infection in men, its potential impact at the population level is difficult to assess. OBJECTIVE: To estimate the prevalence of Chlamydia trachomatis in young men in clinic and community based samples in north west London. METHOD: Cross sectional survey in healthcare centres and general practices in north west London. 1002 males aged 18-35 years, living in north west London, were recruited by staff in occupational health departments, general practices, student health services, and a "well man" clinic and by postal recruitment in four GP practices. The men were tested for C trachomatis using the ligase chain reaction assay on urine samples. The main outcome measure was prevalence of C trachomatis infection in men aged 18 35 years. RESULTS: The overall response rate was 51%. Prevalence of confirmed infection was 1.9% (95% CI: 1.14% to 2.96%) in all men. Best estimated minimum prevalence of infection was 1% (95% CI: 0.58% to 1.50%). Estimated prevalence was highest among men aged over 30 years. CONCLUSIONS: The estimated prevalence among men is commensurate with that described for female populations in London. The results suggest that recruitment of men to screening programmes would be difficult. However, a higher proportion of chlamydial infection may be detected in men than in women by existing approaches to control through genitourinary medicine clinic based case finding and contact tracing. Screening of young women and the contact tracing of the male partners of positive females may be an efficient approach to improving chlamydia control. PMID- 11026884 TI - Herpesvirus infection of eye and brain in HIV infected patients. AB - OBJECTIVES: To compare histological with genome detection methods for diagnosis of herpesvirus infection in eye and brain of HIV infected patients undergoing necropsy and to correlate these findings with both antemortem clinical findings and postmortem evidence of extraocular herpesvirus infection, especially in the CNS. METHODS: A prospective study of 31 consecutive HIV infected patients undergoing necropsy. In life 11 patients had been assessed by an ophthalmologist because of ocular symptoms. Ocular and brain samples were examined for herpesviruses by conventional histological methods and by nested polymerase chain reaction (nPCR) for all eight human herpesviruses; evidence of extraneural herpesvirus infection was sought by histological methods. RESULTS: Although only 12 out of 31 patients (39%) had antemortem clinical evidence of ocular or CNS herpesvirus associated disease, herpesviruses were detected by nPCR in eye and brain from 26 (84%) patients; six patients had more than one herpesvirus infection. There was concordance between ocular and CNS findings in 15 of 19 patients (79%) with CMV infection. 17 of 31 patients (55%) had extraocular or CNS CMV infection at necropsy. Genome detection using nPCR was superior to histological methods for diagnosis of ocular and CNS herpesvirus infection. CONCLUSION: Herpesvirus infection of eye and brain was a frequent finding at necropsy in this group of HIV infected patients; almost a fifth were co-infected by more than one herpesvirus. This was more than twice the incidence predicted from clinical evidence before death. Genome detection using nPCR was superior to histological methods for diagnosis of ocular and CNS herpesvirus infection. PMID- 11026883 TI - The power of information and contraceptive choice in a family planning setting in Mexico. AB - OBJECTIVES: This study measured the effect of information about family planning methods and STD risk factors and prevention, together with personal choice on the selection of intrauterine devices (IUDs) by clients with cervical infection. METHODS: We conducted a randomised, controlled trial in which family planning clients were assigned to one of two groups, the standard practice (control) group in which the provider selected the woman's contraceptive and the information and choice (intervention) group. The study enrolled 2107 clients in a family planning clinic in Mexico City. RESULTS: Only 2.1% of the clients had gonorrhoea or chlamydial infections. Significantly fewer women in the intervention group selected the IUD than the proportion for whom the IUD was recommended in the standard care group by clinicians (58.2% v 88.2%, p = 0.0000). The difference was even more pronounced among infected women: 47.8% v 93.2% (intervention v control group, p = 0.0006). CONCLUSIONS: The intervention increased the selection of condoms and reduced the selection of IUDs, especially among women with cervical infections, for whom IUD insertion is contraindicated. PMID- 11026885 TI - Neonatal herpes prevention: a minor public health problem in some communities. AB - BACKGROUND: Neonatal herpes is a condition with high morbidity and mortality. The greatest risk occurs when the mother acquires herpes simplex virus (HSV) towards the end of pregnancy. A study from Seattle has suggested that the risk of acquisition of HSV during pregnancy was 3.7%. In Australia, HSV-2 infection is less common in pregnant women than in the United States. Consequently we conducted a study to establish HSV seroprevalence and the rate of HSV seroconversion in this population. METHODS: The study was conducted at Westmead Hospital, Sydney, between June 1995 and April 1998. Women completed a questionnaire covering risk factors for the acquisition of genital herpes. A serum sample during pregnancy and a specimen of cord blood were obtained and tested for antibodies to HSV-2 using a type specific indirect enzyme linked immunosorbent assay (ELISA). Equivocal results were resolved by western blot. A subset of the paired sera was tested for antibodies to HSV-1. The data were analysed using SPSS. RESULTS: 326 of the 2616 (12.5%) women were HSV-2 seropositive. Three women (0.15%) acquired HSV-2 infection during pregnancy. None of the three babies of these mothers developed neonatal herpes. 416 maternal cord pairs were tested for HSV-1 antibodies and 330 (79.3%) were positive. No HSV-1 seroconversions occurred. CONCLUSIONS: In this population, HSV acquisition was uncommon (0.34% per year) and neonatal herpes was rare. A cost effective analysis suggested that type specific serology to screen pregnant women and their partners in low prevalence communities was not cost effective. PMID- 11026886 TI - Health issues associated with increasing use of "crack" cocaine among female sex workers in London. AB - OBJECTIVES: To document changes in "crack" cocaine use in the sex industry in London, and to assess health risks associated with the drug. DESIGN: Two serial cross sectional surveys. SUBJECTS: Sex workers interviewed in 1989-9 and 1995-6. MAIN OUTCOME MEASURES: Self reported use of crack cocaine; clinical history of sexually transmitted infection and pregnancy, clinical outcomes. RESULTS: The proportion of women reporting crack use increased significantly from 22/193 (11%) in 1989-91 to 48/143 (34%) in 1995-6. Women in all the main prostitution sectors reported crack use. Crack users had been working in prostitution for longer, were more likely to have worked on the streets, to inject drugs, and to have a partner who injected. Crack use was associated with termination of pregnancy and with hepatitis C infection. The association with hepatitis C was partially explained by confounding with injecting drug use. CONCLUSIONS: Crack use is more common and less problematic than clinical presentation suggests. Use has increased over the past decade, and is associated with hepatitis C infection and termination of pregnancy. It is possible that crack use facilitates hepatitis C transmission due to oral lesions from smoking. Crack use can be difficult to identify because of the stigma of being labelled a "crack whore," therefore information on crack might usefully be integrated into general health promotion material on drugs and safer sex. PMID- 11026887 TI - HIV, sexually transmitted infections, and risk behaviours in male commercial sex workers in Sydney. AB - OBJECTIVES: To assess prevalence of HIV and sexually transmitted infections (STIs), risk behaviours, and demographics in male commercial sex workers (CSWs)/prostitutes in Sydney. METHODS: Retrospective, cross sectional study with two comparison groups. Demographic, behavioural, and morbidity data were analysed from standardised medical records of patients attending a public STI and HIV service in Sydney between January 1991 and March 1998. Two comparison groups were used: female CSWs and non-CSW working homosexual men who attended over the same time. RESULTS: 94 male CSWs, 1671 female CSWs, and 3541 non-CSW working homosexual men were included. The prevalence of HIV in male CSWs tested (6.5%) was significantly greater than in female CSWs (0.4%, p = 0.0001), but less than in non-CSW homosexual men (23.9%, p < 0.0001). Genital warts occurred significantly more frequently in male CSWs than in comparison groups. Prevalence of other STIs was similar in all groups. Male CSWs saw significantly fewer clients per week than female CSWs and male and female CSWs used condoms with almost all clients. Male CSWs reported significantly more non-work sexual partners than female CSWs and non-CSW homosexual men and were significantly more likely to have unprotected penetrative sex with their non-work partners than non CSW homosexual men. Injecting drug use was significantly more frequent in male CSWs than in both comparison groups. CONCLUSIONS: Although male CSWs use condoms with clients, they are more likely to practise unsafe sex with non-work partners (especially women) and inject drugs than female CSWs and non-CSW homosexual men. Some men with HIV are working within the commercial sex industry. Targeted health education to encourage safer drug use and safer sex outside work is needed. PMID- 11026888 TI - STD management by private pharmacies in Hanoi: practice and knowledge of drug sellers. AB - BACKGROUND: Prompt treatment of sexually transmitted infections may reduce the incidence of HIV/AIDS infections. With health sector reforms private pharmacies are increasingly the first and only contact with health delivery services. OBJECTIVES: To find out how patients with STDs are treated at private pharmacies in Hanoi, and what drug sellers know about STD management. METHODS: Five simulated clients were taught to adopt a scenario stating that their friend had a urethral discharge. They visited 60 randomly selected private pharmacies in urban Hanoi and noted all questions asked, advice offered, and treatment given. Afterwards interviewers administered a semistructured questionnaire to all people working in the 60 pharmacies. RESULTS: Drug treatment was given in 84% of the 297 encounters averaging 1.5 drugs and 1.2 antibiotics per encounter. Quinolones were given 188 times. No dispensing was adequate for chlamydia or was in accordance with the national guidelines. No questions were asked in 55% of encounters and no advice was given in 61%. Questions on sexual activity were asked in 23% (69) of cases and about the health of the partner twice (1%). Advice to practise safe sex was given in 1% of encounters and for the partner to seek treatment only once. Of 69 questionnaires administered 51% said they would refer to a doctor, 16% said they would ask about the sexual activity 1% said they would ask about the health of the partner, 7% said they would advise using a condom, and 1% advised telling the partner to seek treatment. Even after prompting, 61% would ask no questions and 80% would give no advice. CONCLUSIONS: Even though 74% of pharmacists and drug sellers know that they should not treat STD patients, 84% actually did. None gave syndromically correct treatment. In both the questionnaire and during the simulated client methods, numbers advising on partner notification and condom use were very poor. Educational or peer awareness interventions are urgently needed among private pharmacists in Vietnam. PMID- 11026889 TI - Syndromic management of vaginal discharge among women in a reproductive health clinic in India. AB - OBJECTIVES: To examine the performance of the syndromic approach in the management of vaginal discharge among women attending a reproductive health clinic in New Delhi, India. METHODS: Women who sought services from the clinic and who had a complaint of vaginal discharge were interviewed, underwent a pelvic examination, and provided samples for laboratory investigations of bacterial vaginosis, candidiasis, syphilis, trichomoniasis, and Chlamydia trachomatis and Neisseria gonorrhoeae infections. Data analysis focused on the prevalence of infection and on the performance of the algorithm recommended by the national authorities for the management of vaginal discharge. RESULTS: The most common infection among 319 women was bacterial vaginosis (26%). At least one sexually transmitted infection was detected in 21.9% of women. The prevalence of C trachomatis infection was 12.2%; trichomoniasis 10%; syphilis 2.2%; N gonorrhoeae was not isolated. An algorithm based on risk assessment and speculum assisted clinical evaluation was not helpful in predicting cervical infections associated with C trachomatis (sensitivity 5% and PPV 9%). This algorithm was sensitive (95%) though not specific (22%) in selecting women for metronidazole therapy effective against bacterial vaginosis or trichomoniasis, and overtreatment was a problem (PPV 38%). The sensitivity, specificity, and PPV of this algorithm for the treatment of candidiasis were 46%, 98%, and 88% respectively. The cost per case assessed using the algorithm was $2 and the cost per infection correctly treated was $4.25. CONCLUSIONS: The prevalence of cervical infection associated with C trachomatis was high among these "low risk" women. The syndromic approach is not an efficient tool for detecting this condition, and alternative approaches to evaluation and intervention are required. The syndromic management of vaginal discharge among women seeking family planning and other reproductive health services should focus on vaginal infections, thus enhancing quality of care and addressing women's concerns about their health. PMID- 11026890 TI - Screening for HIV infection in genitourinary medicine clinics: a lost opportunity? British Co-operative Clinical Group. AB - OBJECTIVES: To examine the policy and practice of HIV testing in genitourinary medicine clinics in the United Kingdom. DESIGN: All 176 consultants in charge of genitourinary medicine clinics in the United Kingdom were sent a policy and practice questionnaire. A self selected group of 53 clinics conducted a retrospective case note survey of the first 100 patients seen in each clinic in 1998. SETTING: Genitourinary medicine clinics in the United Kingdom. SUBJECTS: Consultants in charge of, and case notes of patients attending, genitourinary medicine clinics. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Number of patients tested for HIV. RESULTS: Consultants' assessments of their rate of HIV testing often exceeded the actual rates of testing in the clinic as a whole. The majority of patients deemed to be at high risk requested an HIV test. The exception were heterosexuals who had lived in sub-Saharan Africa. Among attenders at high risk of HIV who did not request a test, 57/196 (29%) were not offered one by clinic staff. Two fifths (51/130) of consultants felt the proportion of patients tested in their clinic was too low. The commonest reason given for this was a lack of time, especially that of health advisers. CONCLUSIONS: A substantial minority of people with HIV infection attending genitourinary medicine clinics fail to have their infection diagnosed. Two major reasons were identified. Firstly, a test was not always offered to those at high risk of HIV. Secondly, a lack of resources, mainly staff, which prevents some clinics from increasing their level of testing. PMID- 11026891 TI - Clinical features of lichen sclerosus in men attending a department of genitourinary medicine. AB - OBJECTIVES: To characterise the clinical presentation and response to treatment of lichen sclerosus (LS) in men attending a department of genitourinary medicine. METHODS: A case note review of all men attending a GUM department over a 32 month period, who had been diagnosed as having LS. RESULTS: 66 men were seen with genital LS. The mean age at diagnosis was 36.5 years but symptoms had been present for up to 10 years before the diagnosis was made. 55 men underwent biopsy and the diagnosis was made histologically in 47 of these men. At the time of presentation 30% of men had no symptoms relating to their LS. All were treated with potent topical steroids and surgery was avoided in nearly all of them. CONCLUSIONS: LS is not uncommon in men presenting to a GUM department, and is often asymptomatic. The disease responds well to potent topical steroids allowing the normal anatomy to be preserved in most individuals. PMID- 11026892 TI - Seroprevalence of syphilis among women attending urban antenatal clinics in Burkina Faso, 1995-8. The DITRAME Study Group. DIminunation de la TRAnsmission Mere-Enfant. AB - OBJECTIVES: To describe annual trends in syphilis seroprevalence and to identify risk factors of syphilis among pregnant women receiving antenatal care in Bobo Dioulasso, Burkina Faso. METHODS: Women were recruited between January 1995 and July 1998 in three antenatal clinics where counselling and HIV testing services had been established in the context of a trial evaluating a short course of zidovudine to reduce mother to child transmission of HIV (ANRS 049 trial). Sociodemographic variables were collected during HIV pretest counselling sessions. Syphilis diagnosis was considered when serum was positive with both rapid plasma reagin and Treponema pallidum haemagglutination assay (TPHA) tests. RESULTS: Overall, 10,980 pregnant women were screened. Syphilis seroprevalence was 0.24% (95% confidence interval (CI): 0.15-0.35) without changes over time. HIV prevalence was 8.8% (CI: 8.3-9.3). In a multivariable analysis, having casual sex partners (odds ratio (OR) = 4.48; CI: 1.62-12.38), being HIV seropositive (OR = 2.62; CI: 1.02-6.74), and being illiterate (OR = 3.78; CI: 1.24-11.48) were independent risk factors for syphilis infection. CONCLUSIONS: This study suggests low syphilis seroprevalence in this city of Burkina Faso. Sexually transmitted disease programmes should be reinforced to offer free access to syphilis screening and treatment in order to eliminate this disease, in coordination with HIV prevention and care. PMID- 11026893 TI - Sexual co-transmission of HIV, hepatitis B, and hepatitis C viruses. PMID- 11026894 TI - Sexually transmitted infections, drug use, and risky sex among female sex workers in Guyana. PMID- 11026896 TI - At last I've been awarded the BTA! PMID- 11026895 TI - A "stone" in the vulva. PMID- 11026897 TI - Lymphatic filariasis--lest we forget. PMID- 11026898 TI - Canary to sparrow; what is in a name? PMID- 11026899 TI - Acceptability of home screening for chlamydial infection: some remaining issues. PMID- 11026900 TI - Nurse counselling for women with abnormal cervical cytology improves colposcopy and cytology follow up attendance rates. PMID- 11026901 TI - Phone sex: information technology (IT) and sexually transmitted infection in young people. PMID- 11026902 TI - Gonorrhoea: an incidence graph of Mersey region data for the 1990s and discussion on the factors behind the changing pattern of incidence. PMID- 11026903 TI - Russian STI. PMID- 11026904 TI - Cheilitis in association with indinavir. PMID- 11026905 TI - Stand-up stuff. PMID- 11026906 TI - Privacy vs. fund-raising. PMID- 11026907 TI - The little picture. PMID- 11026908 TI - Outpatient prospective payment. A long-awaited [yet dreaded] debut. PMID- 11026909 TI - Medical telemetry. Channel surfing. PMID- 11026910 TI - Risk management in California. Riskier than ever. PMID- 11026911 TI - Flu preparedness. Germ warfare. PMID- 11026912 TI - Needle safety. Sticking point. PMID- 11026913 TI - Club ED: emergency room with hotel flair. PMID- 11026914 TI - Alzheimer's care. 'Windows to the heart'. PMID- 11026915 TI - 10th annual compensation salary guide. PMID- 11026916 TI - Six forces shaping managed care. PMID- 11026917 TI - Election 2000. Health care takes center stage. PMID- 11026918 TI - Advances in diagnostic and therapeutic endoscopy. AB - Endoscopy is extremely valuable in the evaluation of disorders of the luminal gastrointestinal tract, pancreas, and biliary system. Endoscopy as a medical discipline continues to evolve and is becoming increasingly therapeutic in nature. Minimally invasive endoscopic intervention now is effective in a wide variety of disorders, including gastrointestinal hemorrhage, obstructive diseases of the intestinal or biliary tree, and early detection or prevention of neoplastic disease of the colon and esophagus. The development of EUS technology has expanded greatly the potential utility of endoscopy as a diagnostic and a therapeutic modality, and further technologic advances are anticipated. PMID- 11026919 TI - Hepatology in the new millennium. Advances in viral hepatitis, hepatic disorders, and liver transplantation. AB - The 1990s have been an exciting time for the field of hepatology. There has been a rapid expansion of knowledge, and new discoveries have revolutionized the field. It is now possible to characterize and treat many more liver diseases. Newer medications in the form of interferon alfa and nucleoside analogues have been added to the armamentarium for treatment of chronic viral hepatitis. Liver transplantation has been established as an effective therapy for patients with end-stage liver disease. PMID- 11026920 TI - Advances in inflammatory bowel disease. AB - Optimal management of patients with IBD requires a multidisciplinary approach involving primary care physicians, gastroenterologists, surgeons, radiologists, and nutritionists. The rapidly evolving medical armamentarium promises better quality of life for patients afflicted with these complex, chronic diseases. It is expected that future development of biologic agents will add to the therapeutic options, although it may complicate treatment algorithms. Surgical advancements, particularly in ileoanal anastomosis and bowel preservation by strictureplasty, have improved outcome dramatically. The focus on development of new therapies and refinement of older ones demands a constant attention to the latest peer-reviewed literature and that the clinician keep abreast of the various advancements that have been summarized here. PMID- 11026921 TI - Helicobacter pylori. AB - H. pylori infection is now recognized as causing serious and life threatening disease in 20% to 30% of those infected. Reliable therapy is problematic. This article addresses the current approach to diagnosis and therapy and new considerations regarding whom to treat. The emphasis of the association of the gastric cancer phenotype of H. pylori infection (cagA positive H. pylori corpus gastritis) and protection against gastroesophageal reflux disease is an example of epidemiology without regard to the biology. Improvements in health and diet and an increase in body mass have accompanied the natural loss of H. pylori from the population. The search for virulence factors to help direct therapy to those who would most benefit has, to date, proved fruitless. Whenever H. pylori is diagnosed, it should be treated. PMID- 11026922 TI - Gastroesophageal reflux disease and Barrett's esophagus. AB - GERD has emerged as an important medical issue by virtue not only of its high prevalence, but also by the concern that it may predispose to adenocarcinoma of the esophagus. It generally is classified into erosive and nonerosive forms. Nonerosive GERD tends to remain as such in most patients, and treatment is based on symptom relief. In contrast, erosive GERD mandates aggressive lifelong treatment based on its inevitable relapse without appropriate treatment. Two excellent therapies are available for GERD. Proton-pump inhibitors are highly efficacious and have an excellent long-term (> 10 years) safety profile. Laparoscopic fundoplication offers a new and easier method of delivering a standard reliable procedure also with excellent long-term efficacy. The occurrence of Barrett's esophagus among reflux patients has emerged as an important problem mostly because of the rapidly rising incidence of adenocarcinoma of the esophagus in the population. All patients with long standing reflux symptoms should be screened for Barrett's and subsequently followed regularly with surveillance endoscopy if Barrett's is detected. Although there are no data to show that aggressive medical or surgical treatment alters the malignant potential of this disease, patients need lifelong therapy. PMID- 11026924 TI - Acute gastrointestinal bleeding. AB - Acute gastrointestinal bleeding is a significant worldwide medical problem. Despite modern measures for diagnosis and treatment, morbidity and mortality rates associated with gastrointestinal bleeding remain largely unchanged. Aggressive medical resuscitation while initiating an evaluation to localize the site of blood loss remains the key to successful management of acute gastrointestinal bleeding. A multidisciplinary approach with early involvement of a gastroenterologist, surgeon, and radiologist can be extremely helpful in the management of these patients. With the logical and direct approach to the evaluation of patients with gastrointestinal bleeding described in this article, most episodes can be managed successfully. PMID- 11026923 TI - Colorectal cancer update. Prevention, screening, treatment, and surveillance for high-risk groups. AB - Great advances have been made in understanding the cause and molecular genesis of colorectal cancer. The disease can be prevented by a healthful diet and lifestyle or by resecting the precursor of most of these cancers, the advanced adenomatous polyp. Screening the average-risk population plus special surveillance for high risk groups now is recommended by evidence-based guidelines. Surgery is highly curative for patients without distant metastases, and adjuvant therapy improves survival in selected patients with advanced cancers. PMID- 11026925 TI - Nutrition. AB - The use of nutrition for the medical patient, in the inpatient setting and at home, will likely continue to increase in the future. Each patient should be evaluated in an individualized but systematic fashion. Each patient in whom malnourishment is suspected should undergo a thorough assessment for the presence and degree of malnutrition with an accurate calculation of nutritional requirements. It is important to choose the correct method of delivery of nutrition, to monitor and recognize any complications or problems that may arise, and to tailor the nutritional therapy to the unique diseases that are encountered in medicine. Although increasingly new advances and changes are occurring in the field of nutrition, nutritional support and therapy are best delivered and supplied to the patient with a network of health care workers, including the physician, the nurse, the dietitian, the social worker, and pharmacist. PMID- 11026927 TI - Irritable bowel syndrome. AB - Patients with IBS need an integrated, individualized approach to treatment, with a foundation based on a successful patient-physician relationship. The emotional and psychologic needs of the patients need to be considered in a successful management plan. Further advances in the pathophysiology of disease are essential to develop suitable and more comprehensive treatment options. PMID- 11026926 TI - Constipation. AB - Most patients with chronic constipation respond to dietary measures, fiber supplements, or laxatives. For those who do not, diagnostic tests of colonic and anorectal function help to select appropriate treatments. Therapeutic options, pharmacologic agents, and surgery, the last resort, are discussed in this article. PMID- 11026928 TI - Diarrhea. AB - Diarrhea is a common symptom of many conditions, and making an accurate diagnosis can be daunting. Research efforts have expanded the differential diagnosis of chronic diarrhea and have provided improved methods for the evaluation and management of patients with diarrheal diseases. This article discusses some of these trends and presents the emerging consensus regarding appropriate pathways of caring for patients with diarrheal diseases. PMID- 11026929 TI - Drug-induced liver disease. AB - The incidence of drug-induced liver disease appears to be increasing, reflecting the increasing number of new agents that have been introduced into clinical use over the past several decades. Among the topics covered, the author discusses incidence, diagnosis, risk factors, clinical presentations, hepatitis, and vascular injury. The author also reviews the hepatic injury seen with commonly prescribed drugs, emphasizing newer developments in the field and recent publications and reports. PMID- 11026930 TI - Psychologic and psychiatric aspects of gastrointestinal disease. AB - There has been an explosion in understanding of the psychosocial concomitants of functional gastrointestinal disorders. Detecting psychologic disturbance and eliciting a history of physical or sexual abuse are critical in suggesting comprehensive and efficacious treatment strategies for these patients. The challenge is to define further the use of psychopharmacologic agents, including the newer antidepressants, anticonvulsants, and anxiolytic agents, in the treatment of chronic functional gastrointestinal disorders. Further research to evaluate the usefulness of various forms of psychotherapeutic and behavioral interventions needs to be undertaken. Establishing a multicomponent treatment program delivered by a team of caregivers, each bringing their unique skills (internist, psychiatrist, psychologist, and others) to patients, must be based on further research on the efficacy of these modalities as opposed to empiric treatment. PMID- 11026931 TI - Nonsteroidal anti-inflammatory drugs. AB - NSAID-associated dyspeptic symptoms are common and can be managed empirically with an H2-receptor antagonist or a proton-pump inhibitor. Treatment of established gastroduodenal ulcers is accomplished best by withholding the offending drugs. Proton-pump inhibitors appear to heal ulcers at the same rate whether or not NSAID therapy is continued. After the ulcer is healed and if NSAID therapy must be continued, prophylaxis is accomplished best by the concomitant use of proton-pump inhibitors, misoprostol (at least 200 micrograms 3 times a day), or a NSAID that preferentially inhibits COX-2. The future development of newer, safer NSAID preparations, including highly selective COX-2 inhibitors and nitric oxide-releasing NSAIDs, should provide better treatment options for the increasing number of individuals requiring anti-inflammatory agents. PMID- 11026932 TI - TSH assay using dried blood samples: a screening method for congenital hypothyroidism. AB - Capillary blood samples from three hundred healthy, full-term newborn within 48 and 120 hours of delivery, were collected on filter paper for analysis of thyroid stimulating hormone (TSH). Blood was collected by pricking the heel of the newborn and spotted and dried on filter paper cards. The samples were analysed by immunoradiometric assay (IRMA) to determine the TSH level. Some fifteen percent of a total 1928 deliveries at two hospitals during a period of one year were included in the study. Male:female ratio was 1.1:1. There was no maternal history of thyroid disease. None of the babies had any clinical feature of hypothyroidism. On analysis, TSH level ranged from 0.6-19.3 microU/ml with a mean (SD) of 7.19 (4.21) microU/ml. TSH assay from dried filter paper blood spot is technically possible and is a cost-effective and reliable method for a screening programme. This is the first time in Bangladesh that this method has been successfully carried out. PMID- 11026933 TI - Limiting factors of ORS intake. AB - This was a thirty cluster survey; carried out to identify the factors which limit ORS intake during a diarrhoeal episode in rural Bangladesh. 420 mother-child pairs were investigated. In addition to the verbal interview, the index child was examined physically and mother's skill of preparing ORS was assessed. The ORS was known to more than 95% mothers but used by only 33.3%. The average intake of ORS was 320 ml per diarrhoea day. The correct amount of oral rehydration fluid was received by 145(34.5%) cases. Twenty eight socio-demographic-maternal-host(child) environmental factors were studied for their probable influence on intake of correct amount of oral rehydration fluid. In bivariate analysis, fourteen factors were found to be associated significantly with oral rehydration fluid intake (P < 0.05-< 0.0001). These fourteen factors were treated with stepwise logistic regression analysis. Sensitivity and specificity were the key measures for decision of best fitted model. A combination of child age, mother's education, socio-economic condition and method of feeding ORS provide the maximum sensitivity (90.20%) and specificity (74.19%) for predicting oral rehydration fluid intake. The study emphasized the need for further strengthening of health education incorporating the findings of the research to ensure required amount of oral rehydration fluid intake during diarrhoeal episode. PMID- 11026934 TI - Humoral immune response in a sample of Bangladeshi children with rheumatic fever. AB - Several researchers have investigated cellular response, and antibody response to different antigens of group A beta-haemolytic streptococci, cardiac tissues and B cell alloantigen in rheumatic fever (RF). However, immunoglobulins (Ig) which may indicate general immune status did not receive attention. This study was done in the outpatient clinic of the National Center for Control of Rheumatic Fever and Heart Diseases, Dhaka, to compare Ig levels in subjects with and without RF who have had recent group A beta-haemolytic streptococcal infections. We have recruited 44 RF cases aged 5 to 20 years, and 44 subjects without RF were randomly matched for age and sex as controls. Convalescent blood samples were used to measure antistreptolysin O and IgG, IgM, and IgA levels. The cases, as compared with the controls, had significantly higher levels of antistreptolysin O (mean 399 versus 321 IU/ml), IgG (mean 2386 versus 1885 mg/dl), IgM (mean 286 versus 222 mg/dl) and IgA (mean 258 versus 184 mg/dl). It is not clearly known why the immune response is higher in the RF cases. We have to elucidate factors responsible for higher immune response in children with RF. PMID- 11026935 TI - Recurrent respiratory papillomatosis: a study of 24 cases. AB - 24 cases of recurrent respiratory papillomatosis (RRP) in children were treated by surgical removal with cup forceps and followed up for variable periods between 1990-95. Most of the patients required multiple operations due to recurrence. No anaesthetic death or complication was recorded. Voice changes of different degrees were noted. Four patients has glottic web formation. The findings suggest that recurrence of RRP is very common & needs multiple session of surgery. For children with persistent and progressive voice change/cry, the necessity of precise & comprehensive endoscopic examination of the upper aerodigestive tract under general anesthesia (GA) is emphasized. PMID- 11026936 TI - TB control in Bangladesh: more challenges ahead. PMID- 11026937 TI - The National Tuberculosis Control Programme. PMID- 11026938 TI - Achievements of the National TB Control Programme. PMID- 11026939 TI - On the viscoelastic character of liver tissue: experiments and modelling of the linear behaviour. AB - This paper presents the results of recent investigations into the viscoelastic character of liver tissue, especially the linear response. The linear domain of liver tissue has been determined by a strain sweep oscillation test. The fluid like viscoelastic behaviour of liver tissue was confirmed by stress relaxation experiments. To demonstrate the consistency and quality of the experimental data, the relaxation spectrum was used to reconstruct the relaxation modulus, as well as the storage and loss moduli. The generalised Maxwell model was successfully employed to model the mechanical behaviour in the linear viscoelastic domain. PMID- 11026940 TI - The influence of temperature and length of time of storage of frog mucus samples. AB - Frozen, stored mucus has been extensively used for transport studies but there is no clear evidence of the influence that the temperature and length of time of storage may have on the results. We stored frog mucus samples at -20 and -80 degrees C and analysed them on days zero, 2, 10, 30 and 90. At each temperature, a sample was thawed, studied and refrozen on each of the study days, at the same time that one sample was thawed only on the study day. Displacement in a simulated cough machine and on the frog palate, as well as contact angle measurements, were determined for the mucus samples on each study day. Mucus cytologic analyses on each of the study days were done with special regard to neutrophil counts and cell integrity. Friedman analysis of variance did not show any difference between the different periods of storage and the two temperatures for any of the parameters studied. The medians for the relative transport velocity on the frog palate varied between 0.88 and 1.03, for the contact angle between 21 and 28 degrees, and for the displacement in the simulated cough machine between 58 and 95 mm over the 90 days of the experiment. There were no cytologic alterations compatible with cell degeneration. We conclude that the storage of frog mucus either at -20 or -80 degrees C for periods up to 90 days does not lead to any significant differences in mucus transportability. PMID- 11026941 TI - Computer simulation of osmotic expansion and shrinkage in okra hypocotyl segments. AB - Okra hypocotyl segments were incubated in solutions of 0.3 or 0.4 M sorbitol at various temperatures and their shrinkage was measured. The result yielded an apparent activation energy for shrinkage of 4.8 kcal/mol, which is close to that of the viscosity of water. This coincidence suggests that the viscosity of water, i.e., the reciprocal function of water conductivity, is a limiting factor for osmotic shrinkage. Abrasion of okra hypocotyl segments with Carborundum substantially increased the rate of their osmotic shrinkage, indicating that the cuticle is the major barrier to water uptake by segments. The apparent activation energy for osmotic shrinkage was 4.5 kcal/mol in abraded segments. By introducing water conductivity into an algorithm, osmotic shrinkage and expansion of hypocotyl segments was successfully predicted by computation with this algorithm. Hence the extent of the contribution of water conductivity in osmotic shrinkage and expansion can be evaluated. Based on this simulation, water conductivity was identified as one of the major factors in governing the elongation growth rate of cells along with the osmotic pressure of the cell sap and the mechanical properties of the cell wall. PMID- 11026942 TI - Detection of red cell aggregation by low shear rate viscometry in whole blood with elevated plasma viscosity. AB - The viscosity of whole blood measured at low shear rates is determined partly by shear resistance of the red cell aggregates present, stronger aggregation increasing the viscosity in the absence of other changes. Effects of cell deformability can confound interpretation and comparison in terms of aggregation, however, particularly when the plasma viscosity is high. We illustrate the problem with a comparison of hematocrit-adjusted blood from type 1 diabetes patients and controls in which it is found the apparent and relative viscosities at a true shear rate of 0.20 s-1 are lower in the patient samples than age matched controls, in spite of reports that aggregation is increased in such populations. Because the plasma viscosities of the patients were higher on average than controls, we performed a series of experiments to examine the effect of plasma protein concentration and viscosity on normal blood viscosity. Dilution or concentration by ultrafiltration of autologous plasma and viscosity measurements at low shear on constant hematocrit red cell suspensions showed (a) suspension viscosity at 0.25 and 3 s-1 increased monotonically with plasma protein concentration and viscosity but (b) the relative viscosity increased, in concert with the microscopic aggregation grade, up to a viscosity of approximately 1.25 mPa-s but above this the value the relative viscosity no longer increased as the degree of aggregation increased in concentrated plasmas. It is suggested that in order to reduce cell deformation effects in hyperviscous pathological plasmas, patient and control plasmas should be systematically diluted before hematocrit is adjusted and rheological measurements are made. True shear rates should be calculated. Comparison of relative viscosities at low true shear rates appears to allow the effects of red cell aggregation to be distinguished by variable shear rate viscometry in clinical blood samples. PMID- 11026943 TI - Computational fluid dynamic studies of leukocyte adhesion effects on non Newtonian blood flow through microvessels. AB - The study of the effect of leukocyte adhesion on blood flow in small vessels is of primary interest to understand the resistance changes in venular microcirculation. Available computational fluid dynamic studies provide information on the effect of leukocyte adhesion when blood is considered as a homogeneous Newtonian fluid. In the present work we aim to understand the effect of leukocyte adhesion on the non-Newtonian Casson fluid flow of blood in small venules; the Casson model represents the effect of red blood cell aggregation. In our model the blood vessel is considered as a circular cylinder and the leukocyte is considered as a truncated spherical protrusion in the inner side of the blood vessel. The cases of single leukocyte adhesion and leukocyte pairs in positions aligned along the same side, and opposite sides of the vessel wall are considered. The Casson fluid parameters are chosen for cat blood and human blood and comparisons are made for the effects of leukocyte adhesion in both species. Numerical simulations demonstrated that for a Casson fluid with hematocrit of 0.4 and flow rate Q = 0.072 nl/s, a single leukocyte increases flow resistance by 5% in a 32 microns diameter and 100 microns long vessel. For a smaller vessel of 18 microns, the flow resistance increases by 15%. PMID- 11026944 TI - George William Scott Blair, MA PhD DSc FRIC FInstP (1902-1987): the man and his work. PMID- 11026945 TI - Fitting the biopsychosocial jigsaw together. PMID- 11026946 TI - The London Depression Intervention Trial. Randomised controlled trial of antidepressants v. couple therapy in the treatment and maintenance of people with depression living with a partner: clinical outcome and costs. AB - BACKGROUND: Relapse of depression is associated with a criticising attitude of the patient's partner. AIMS: To compare the relative efficacy and cost of couple therapy and antidepressant drugs for the treatment and maintenance of people with depression living with a critical partner. METHOD: A randomised controlled trial of antidepressant drugs v. couple therapy. The subjects were 77 people meeting criteria for depression living with a critical partner. RESULTS: Drop-outs were 56.8% [corrected] from drug treatment and 15% from couple therapy. Subjects' depression improved in both groups, but couple therapy showed a significant advantage, according to the Beck Depression Inventory, both at the end of treatment and after a second year off treatment. Adding the costs of the interventions to the costs of services used showed there was no appreciable difference between the two treatments. CONCLUSIONS: For this group couple therapy is much more acceptable than antidepressant drugs and is at least as efficacious, if not more so, both in the treatment and maintenance phases. It is no more expensive overall. PMID- 11026947 TI - Cognitive-behavioural techniques for general psychiatrists in the management of patients with psychoses. AB - BACKGROUND: Recent research progress showing the benefits of cognitive therapy in schizophrenia leaves the general psychiatrist unsure whether to attempt to use such techniques. AIMS: To test whether cognitive-behavioural techniques are beneficial in the management of patients with schizophrenia in general psychiatric practice. METHOD: A randomised controlled study comparing the use of cognitive-behavioural techniques and befriending in schizophrenia. RESULTS: Significant improvement in symptoms occurred in the group treated with cognitive behavioural techniques but not in the befriending group. During the 6-month follow-up period the cognitive-behavioural group tended to have shorter periods in hospital. CONCLUSIONS: General psychiatrists could help their patients with schizophrenia by using cognitive-behavioural techniques. Such techniques are well within the capability of general psychiatrists, but their application would involve more of the consultant's time spent in direct contact with patients with psychoses. PMID- 11026948 TI - Early termination of treatment in personality disorder treated in a psychotherapy hospital. Quantitative and qualitative study. AB - BACKGROUND: Factors underlying premature discontinuation of psychosocial in patient treatment are still unclear. AIMS: Investigation of early discontinuation of specialised in-patient psychosocial treatment in a sample of people with personality disorder. METHOD: Out of 134 consecutive admissions to the Cassel Hospital, 42 early drop-outs and 92 patients who remained were compared on demographic and clinical variables. Early drop-outs were invited for in-depth interviews, to explore their hospital experiences. RESULTS: The early drop-out group and the group which remained showed significant differences in occupational status, borderline personality disorder (BPD) and the treatment programme to which they were allocated. All three independent variables predicted early discontinuation of treatment. The qualitative analysis of interview transcripts identified significant problems in institutional dimensions. CONCLUSIONS: Important subjects and process variables contributing to early drop-out in people with personality disorder were identified, with potential implications for clinical practice. PMID- 11026949 TI - Psychotherapies in psycho-oncology. An exciting new challenge. AB - BACKGROUND: There is ample scope to devise forms of psychotherapy in consultation liaison psychiatry, including the newly evolving area of psycho-oncology. AIMS: To highlight the development of psychotherapy in psycho-oncology, providing two illustrations. METHOD: We report on conceptual and clinical research in the context of oncology and palliative care, focusing on (a) an approach for families at risk of maladaptive bereavement; and (b) a group programme for women newly diagnosed with early-stage breast cancer. RESULTS: We were able to introduce new forms of psychological treatment for specific clinical groups, and anecdotal evidence points to useful benefits for participants. CONCLUSIONS: Psychotherapists should grasp the opportunity to bring their skills to the medical arena, but need to subject newly devised interventions to well-designed and methodologically rigorous research. PMID- 11026950 TI - A neurobiologically informed perspective on psychotherapy. AB - BACKGROUND: Polarisation of biological and psychosocial aspects of psychiatry has promoted a form of Cartesian dualism. Current knowledge of the interaction between biology and psychology makes it possible to consider a truly integrative approach to treatment. AIMS: The aim of this overview is to consider conceptual models of how psychotherapy may affect the brain. METHOD: The literature discussing the mutual influence of genes and environment is surveyed. Relevant data involving the influence of psychotherapy on the brain are also reviewed. RESULTS: Research findings suggest that the brain responds to environmental influence through the alteration of gene expression; that psychotherapy has specific measurable effects on the brain; and that implicit memory may be modified by psychotherapeutic interventions. CONCLUSIONS: Advances in neuroscience research have led to a more sophisticated understanding of how psychotherapy may affect brain functioning. These developments point the way towards a new era of psychotherapy research and practice in which specific modes of psychotherapy can be designed to target specific sites of brain functioning. PMID- 11026951 TI - Measurement and psychotherapy. Evidence-based practice and practice-based evidence. AB - BACKGROUND: Measurement is the foundation of evidence-based practice. Advances in measurement procedures should extend to psychotherapy practice. AIMS: To review the developments in measurement relevant to psychotherapy. METHOD: Domains reviewed are: (a) interventions; (b) case formulation; (c) treatment integrity; (d) performance (including adherence, competence and skillfulness); (e) treatment definitions; (f) therapeutic alliance; and (g) routine outcome measurement. RESULTS: Modern methods of measurement can support 'evidence-based practice' for psychological treatments. They also support 'practice-based evidence', a complementary paradigm to improve clinical effectiveness in routine practice via the infrastructure of Practice Research Networks (PRNs). CONCLUSIONS: Advances in measurement derived from psychotherapy research support a model of professional self-management (practice-based evidence) which is widely applicable in psychiatry and medicine. PMID- 11026952 TI - Psychotherapy for patients with complex disorders and chronic symptoms. The need for a new research paradigm. AB - BACKGROUND: A clear distinction has been made between efficacy and effectiveness in relation to the methods of evaluation of new psychological treatments in psychiatry. Efficacy trials target patients with relatively pure conditions, who may not be representative of the patients who are usually referred for psychological treatment in a clinical setting. Few studies have explored the benefits of psychotherapy in patients with complex disorders and enduring symptoms. AIMS: To explore the rationale for the distinction between efficacy and effectiveness, particularly in relation to outcome studies of patients with complex and enduring disorders. METHOD: A narrative review with examples drawn from the literature, and an illustration of a recent naturalistic outcome study which combines features of both efficacy and effectiveness. RESULTS: Studies of patients with complex and mixed disorders can be designed so that they retain internal validity, but also have external validity and are relevant to clinical practice. CONCLUSION: Studies which evaluate psychological interventions should be carried out in populations of patients clinically representative of those who are likely to receive the intervention, should it be shown to be of benefit. PMID- 11026953 TI - Effectiveness of psychotherapeutic treatment of personality disorder. AB - BACKGROUND: Psychiatrists have been criticised for failing to develop adequate treatment for personality disorder. Psychotherapeutic treatments are promising, but their effectiveness is uncertain. AIMS: To investigate the evidence for effectiveness of psychotherapeutic treatment for personality disorder. METHOD: Systematic literature review. RESULTS: There is evidence for the effectiveness of psychotherapy for personality disorder. Problems of case identification, comorbidity, randomisation, specificity of treatment and outcome measurement are inadequately addressed. Authors mainly relied on cohort studies. Evidence neither suggests superiority of one type of therapy over another nor indicates which subgroups of patients should be offered psychotherapy as in-patient, day patient, or out-patient. CONCLUSIONS: New research strategies are needed to show that personality change is both measurable and clinically meaningful. Effectiveness studies using randomised controlled designs are required. The literature suggests that effective treatment should be long-term, integrated, theoretically coherent and focused on compliance. PMID- 11026954 TI - Psychological therapies for post-traumatic stress disorder. AB - BACKGROUND: After exposure to traumatic stressors, a subgroup of survivors (20 30%) will develop post-traumatic stress disorder (PTSD). AIMS: Since the incidence and prevalence rates for PTSD in the community are significant, it is important that general practitioners and psychiatrists be familiar with possible therapeutic options. In this review we shall look at the published evidence about the effectiveness of psychological treatments for PTSD. METHOD: The psychopathological mechanisms involved in PTSD are discussed. Studies of the effectiveness of different psychological therapies are reviewed. RESULTS: The review suggests that persistent fear or shame reactions are key aspects of PTSD. Evidence from systematic reviews suggests that psychotherapeutic treatments are effective in the therapy of reactions based on fear, and may increase the effectiveness of pharmacological therapy. There is less systematic evidence for the efficacy of interventions for symptoms based on shame. CONCLUSIONS: Although a proportion of patients with complex or chronic PTSD may require specialist interventions, most patients can be treated effectively by a general psychiatric service which can offer both pharmacological and psychological interventions. PMID- 11026956 TI - Long-term survival, place of death, and death certification in clinically diagnosed pre-senile dementia in northern England. Follow-up after 8-12 years. AB - BACKGROUND: Information on survival and cause of death in pre-senile dementia is scarce and the organisation of services controversial. AIMS: To study survival, place of death and death certification in pre-senile dementia. METHOD: Patients aged 45-64 were identified from hospital and community sources in the Northern health region (1985-89) and classified as having pre-senile dementia of Alzheimer type (PDAT) or pre-senile vascular dementia (PVD) by applying an algorithm to case notes. Deaths were ascertained from the National Health Service Central Registry (NHSCR) to 31 December 1998. Survival analysis was performed using the SPSS/PC program, and expected survival calculated from life tables. RESULTS: Median survival time from diagnosis was 6.08 years and did not differ significantly in PDAT and PVD, or by age or gender; 19.3% of deaths occurred at home, 24.5% in nursing or residential homes and 56.3% in hospital; 72.4% of the death certificates mentioned dementia or Alzheimer's disease; 15.4% were still alive. CONCLUSIONS: Pre-senile dementia has a variable but usually chronic course, requiring appropriate planning and services. PMID- 11026955 TI - Definition and prevalence of severe and persistent mental illness. AB - BACKGROUND: There is little consistency in how severe mental illness (SMI) is defined in practice, and no operational definitions. AIMS: To test two operationalized definitions, based on the National Institute of Mental Health (1987) definition: the first uses three criteria (diagnosis of psychosis; duration of service contact > or = 2 years; GAF score < or = 50), the second only the last two. METHOD: Annual prevalence rates of SMI in two European catchment areas for each criterion and the criteria combined were calculated. RESULTS: The first definition produced rates of 2.55 and 1.34/1000 in London and Verona, respectively; the second permitted an additional 0.98/1000 non-psychotic disorders to be included in Verona. CONCLUSIONS: The three-dimensional definition selects a small group of patients with SMI who have psychotic disorders. The two dimensional approach allows estimates of SMI prevalence rates which include all forms of mental disorder. PMID- 11026957 TI - Selective serotonin reuptake inhibitor treatment in the UK: risk of relapse or recurrence of depression. AB - BACKGROUND: Patients with depression are often not prescribed antidepressants for an adequate period of time. AIMS: The impact of antidepressant prescribing patterns on the risk of relapse or recurrence of depression is examined. METHOD: The MediPlus UK Primary Care Database was used to identify patients treated for depression with a selective serotonin reuptake inhibitor (SSRI). Records were used to construct hierarchical prescription patterns (less than 120 days, switching/augmentation, upward titration, or stable use) as indicators for the occurrence of relapse or recurrence of depression. RESULTS: Patients with stable use experienced the lowest risk of relapse or recurrence. Factors significantly associated with increased risk include prior use of anxiolytic medications, more comorbid conditions and younger age. CONCLUSIONS: The SSRI prescription pattern most consistent with recommended depression treatment guidelines was associated with the lowest risk of relapse or recurrence. PMID- 11026958 TI - Interhemispheric asymmetry of motor cortical excitability in major depression as measured by transcranial magnetic stimulation. AB - BACKGROUND: Neuroimaging studies of major depressive disorder (MDD) indicate interhemispheric differences in prefrontal cortical activity (right greater than left). AIMS: To investigate whether there are any interhemispheric differences of motor cortical excitability in MDD. METHOD: Eight patients with treatment refractory MDD off medication were assessed for the severity of their depression, and transcranial magnetic stimulation studies (bilateral motor threshold and paired-pulse studies) were conducted. Eight normal controls were also studied. RESULTS: MDD patients showed significant interhemispheric differences in motor threshold and paired-pulse curves, both of which showed lower excitability on the left hemisphere. Such differences were absent in controls. CONCLUSIONS: Our findings may aid the further understanding of the neurophysiology underlying MDD. PMID- 11026960 TI - Hospitalisation and adolescent anorexia nervosa. PMID- 11026961 TI - Possible causes of catatonia in autistic spectrum disorders. PMID- 11026959 TI - Effects of chewing betel nut (Areca catechu) on the symptoms of people with schizophrenia in Palau, Micronesia. AB - BACKGROUND: Although millions of people with schizophrenia live in betel chewing regions, the effects of betel chewing on their symptoms are unknown. Betel nut alkaloids include potent muscarinic cholinomimetics: recent research suggests that these agents may be therapeutic in schizophrenia. AIMS: To compare the primary and extrapyramidal symptom profiles and substance-using habits of betel chewing v. non-chewing people with schizophrenia. METHOD: A cross-sectional study of 70 people with schizophrenia. Symptom ratings measured by the Positive and Negative Syndrome Scale (PANSS) and Extrapyramidal Symptom Rating Scale (ESRS), and demographic and substance-use data, were compared for 40 chewers and 30 non chewers of betel nut. RESULTS: Betel chewers with schizophrenia scored significantly lower on the positive (P = 0.001) and negative (P = 0.002) sub scales of the PANSS than did non-chewers. There were no significant differences in extrapyramidal symptoms or tardive dyskinesia. CONCLUSIONS: Betel chewing is associated with milder symptomatology and avoidance of more harmful recreational drugs. These initial results indicate that longitudinal research is merited. PMID- 11026962 TI - In-patient detoxification after GHB dependence. PMID- 11026963 TI - Suicide in psychiatric hospital in-patients in Ireland. PMID- 11026964 TI - Low blood pressure and depression in the elderly. PMID- 11026965 TI - Complex medical roles in mental health review tribunals. PMID- 11026966 TI - Forensic trials inform the present and future. PMID- 11026967 TI - Involuntary out-patient commitment and supervised discharge. PMID- 11026968 TI - Chronic glaucoma: definition of the phenotype. PMID- 11026969 TI - Molecular genetics of primary congenital glaucoma. AB - Molecular genetic studies conducted during the last several years have thrown some light on the basic molecular defects in primary congenital glaucoma (PCG) and the rationale behind the clinical and genetic presentation of this paediatric eye condition. The existence of a hereditary form of PCG segregating as an autosomal recessive trait with high penetrance is now confirmed. The primary molecular defect underlying the majority of PCG cases has been identified as mutations in the cytochrome P4501B1 (CYP1B1) gene. This gene is expressed in tissues of the anterior chamber angle of the eye. Molecular modelling experiments suggest that mutations observed in PCG patients interfere with the integrity of the CYP1B1 molecule as well as its ability to adopt a normal conformation and bind haem. On the basis of these observations, we hypothesised that CYP1B1 participates in the normal development and function of the eye by metabolising essential molecules that are perhaps used in a signalling pathway. Revealing the identity of this molecule is our major objective since it can lead to as yet unknown biochemical cascades controlling the terminal stages of anterior chamber angle development. PMID- 11026970 TI - The genetics of open-angle glaucoma: the story of GLC1A and myocilin. AB - A linkage analysis study was performed on a single large family with juvenile onset primary open-angle glaucoma (POAG). This led to the recognition that there was a region of chromosome 1q that harboured a gene for juvenile-onset POAG. This chromosomal site was called GLC1A. It was discovered that a gene that produces the protein myocilin resides within this interval and that mutations in myocilin caused most cases of autosomal dominant juvenile-onset POAG. More importantly myocilin mutations also cause up to 4.6% of cases of adult-onset POAG. The prevalence of myocilin mutations is similar regardless of race or geographic location. There are widely variable glaucoma phenotypes depending on the specific mutation in myocilin. Myocilin is expressed in multiple tissues throughout the eye and in many other organs. In the trabecular meshwork the production of myocilin can be induced by the application of topical corticosteroids. The exact function of myocilin in health and disease remains a mystery. PMID- 11026971 TI - Optic nerve head structure in glaucoma: astrocytes as mediators of axonal damage. AB - Increased intraocular pressure (IOP) is recognised as the principal risk factor for the development of glaucomatous cupping of the optic disc. The hypothesis that it disrupts the function of retinal ganglion cell axons by increasing mechanical forces on the lamina cribrosa of the optic nerve head has received considerable experimental support. However, many patients with glaucoma will have progressive cupping even though the IOPs remain within the normal range, suggesting that mechanical compression is unlikely to be the sole cause of optic nerve damage. Clinical studies have emphasised the role of other factors, such as optic nerve head ischaemia, in generating optic disc cupping. One of the outstanding problems in understanding optic nerve head dysfunction in glaucoma has been the elucidation of the pathways that could integrate the effects of IOP and ischaemia to generate the characteristic changes seen. This review considers the role that optic nerve head astrocytes might play in the initiation of axon damage, based on the hypothesis that these cells are sensitive to mechanical or ischaemic factors and are important for the maintenance of retinal ganglion physiology. It discusses their role in the remodelling of the structure of the lamina cribrosa and the effect that this might have on axon function. Recent evidence has shown that the modulation of astrocyte activity, for example by the reduction of the production of nitric oxide, may prevent retinal ganglion cell death in ocular hypertension. The possibility that astrocyte-axon interactions are important in the development of glaucomatous optic neuropathy suggests new avenues of therapeutic intervention, not related to the control of IOP, that would prevent retinal ganglion cell death in glaucoma. PMID- 11026972 TI - Anterior optic nerve microvascular changes in human glaucomatous optic neuropathy. AB - The microvascular changes in the anterior optic nerve in human glaucomatous eyes were examined by selective methylmethacrylate microvascular corrosion castings following cannulation of the central retinal artery and posterior ciliary arteries in 11 normal eyes and 9 glaucomatous eyes. The resulting castings were examined with scanning electron microscopy. Microvascular changes were found in the anterior optic nerves of all the glaucomatous eyes with visual function loss. These findings include areas of capillary filling defects within the anterior optic nerve and a decreased numbers of feeding arteriolar vessels to the anterior optic nerve. In the prelaminar and laminar regions, the typical capillary patterns are lost and laminar striations are not present. Juxtapapillary choroidal and retinal avascular areas were also identified in two of the glaucomatous eyes. Selective microvascular corrosion casting is an excellent method to examine the three-dimensional microvasculature of the anterior optic nerve. Microvascular changes in the anterior optic nerve may play a role in the development of glaucomatous optic neuropathy. PMID- 11026973 TI - Imaging the optic nerve and ganglion cell layer. AB - New imaging techniques have extended the limitations of visualisation of the structures of the optic nerve and fundus in the living human eye. Quantitative reconstruction of depth of the optic nerve head is becoming widely available through the use of the confocal scanning laser ophthalmoscope (cSLO). A separate method of quantitative depth reconstruction is possible using stereo pairs which is not subject to the same artefacts as the tomographic reconstruction. We are investigating the use of these techniques both with conventional stereo imaging and using stereo pairs derived from cSLO images. In these we find that there is additional structural detail evident in the base of the optic nerve head. Further studies made with an instrument optimised to image this region show that this corresponds to the lamina cribrosa. Current work is aimed at further extending the limits of imaging using techniques based on optical coherence tomography, which provides additional depth resolution. Results to date with a prototype device show an improvement of approximately a factor of 10 in depth resolution and that some separate layers of the retina may be visualised including the photoreceptor layer. The goal is to extend these limits to allow visualisation of the ganglion cell layer. PMID- 11026975 TI - In glaucoma, should enthusiasm about neuroprotection be tempered by the experience obtained in other neurodegenerative disorders? AB - Some in vitro and in vivo evidence, as well as rare observations in human eyes with glaucoma, suggests that retinal ganglion cells could be lost by apoptosis during the course of glaucomatous optic neuropathy. There exist also observations indicating that in the vitreous of patients with glaucoma it is possible to measure an increased concentration of glutamate (an excitotoxic amino acid known to induce neuronal apoptosis in animal models). These observations, among others, suggest the possibility of an excitotoxicity mechanism in the pathogenesis of glaucoma and as a consequence the potential for a neuroprotective approach to treating this disorder. Amazingly, not only in glaucoma but also in other neurodegenerative disorders (Parkinson's disease, amyotrophic lateral sclerosis, stroke, etc.) it has been postulated that neurons could be lost through an excitotoxic mechanism. In these non-glaucomatous disorders, quite a large number of clinical trials have already been conducted to determine the potential benefit of different neuroprotective therapies. Unfortunately, with a few rare exceptions, the results of these clinical studies have been very disappointing (in contrast to encouraging results obtained in preclinical trials). The experience acquired in other neurodegenerative disorders should probably be kept in mind when addressing the question of neuroprotection in glaucoma. In particular, the hope raised by preclinical studies showing that drugs could have a beneficial effect on the survival of retinal ganglion cells should certainly be tempered until such an effect is confirmed by clinical trials conducted in patients with glaucoma. PMID- 11026974 TI - 5-Hydroxytryptamine1A agonists: potential use in glaucoma. Evidence from animal studies. AB - Various classes of compounds exist to lower intraocular pressure (IOP) in the treatment of glaucoma. None of them is ideal since some patients respond better than others and the side effects vary between individuals. New classes of compounds need to be introduced to allow the clinician greater scope for effective treatment of all patients. It is now generally agreed that the cause of ganglion cell dysfunction in glaucoma is likely to be multifactorial and that concentrating solely on reducing IOP is inadequate. Irrespective of the reason for the dysfunction, the future goal must be to attenuate cell death. This may be achieved with drugs that interact with components of the retina, and is termed 'neuroprotection'. Thus, drugs that can both reduce IOP and act as neuroprotectants would be ideal for the treatment of glaucoma. In this article we summarise studies on animals which show serotonergic 5-HT1A agonists to both reduce IOP when topically applied to the rabbit eye and blunt the damaging effect to the rat retina and ganglion cells induced by glutamate toxicity or ischaemia. Reduction of IOP occurs via stimulation of 5-HT1A receptors associated with the ciliary processes. Neuroprotection of retinal neurones appears to involve the interaction of 5-HT1A agonists with membrane sodium channels and/or 5-HT1A or even possibly 5-HT7 receptors. Various 5-HT1A agonists are used in patients to treat depression, so classes of these drugs have a proven safety profile for use in patients. The animal studies summarised in this article suggest that 5-HT1A agonists need to be considered as a new class of drugs for the treatment of glaucoma. PMID- 11026976 TI - Vasoprotection as neuroprotection for the optic nerve. PMID- 11026978 TI - Uveoscleral outflow. PMID- 11026977 TI - How does resetting intraocular pressure help optic nerve function? PMID- 11026979 TI - Repair in the rabbit outflow system. AB - An in vivo study was conducted to study repair processes in the injured rabbit outflow system. A uniform injury was produced by raising intraocular pressure (IOP) manometrically to 70 mmHg for 1 h. The recovery process, which was followed clinically for 8 weeks and morphologically for 6 weeks, led to the re establishment of normal meshwork architecture within this period. The morphological studies included light microscopy, autoradiography and electron microscopy. The initial lesion consisted of large deficits in the meshwork with breakdown of cell-to-cell connections, loss of extracellular materials and disruption of the vessels of the aqueous plexus. There was a significant lowering of IOP in the first week of recovery, which thereafter climbed back to normal. Also in the first week the meshwork became infiltrated with inflammatory cells which cleared by 4 weeks. There was some meshwork cell death by either necrosis or apoptosis. The majority of meshwork cells became activated within the first few days and remained activated for at least the first 2 weeks. Tritiated proline incorporation was maximal between 1 and 2 weeks. Tritiated thymidine labelling was seen throughout, but only after the inflammation subsided was it clear that meshwork cells in all regions of the meshwork were proliferating. Our study provided no evidence that normal meshwork cells have a basal proliferative turnover level. Our injury model involved complete repair of the outflow tissues and that required meshwork cells to become activated, mobilise, undertake synthetic activity and proliferate. This is the first example, other than argon laser trabeculoplasty, where meshwork cells in vivo have been induced to divide. Possible therapeutic implications for glaucoma are discussed. PMID- 11026980 TI - Regulation of TIGR/MYOC gene expression in human trabecular meshwork cells. AB - Glucocorticoid (GC) treatment of human trabecular meshwork (HTM) cells produces delayed, progressive cellular and extracellular protein/glycoprotein inductions with characteristics matching those for intraocular pressure elevation with corticosteroid eyedrops. The cloning of the Trabecular Meshwork Inducible Glucocorticoid Response (TIGR) gene from this system has suggested possible environmental and genetic influences in relation to glaucoma mechanisms. As reported here, the major GC-induced increase of TIGR expression in HTM cells is reduced approximately 4-fold by basic fibroblast growth factor (bFGF, 100-1000 pM), with a somewhat smaller inhibition noted with the thyroid hormone triiodothyronine (T3, 100 nM). Such endogenous 'protective' factors could help balance stimulatory effects on TIGR gene expression from 'stress' and/or mechanical perturbations in the trabecular meshwork. TIGR coding region mutations affecting the gene's olfactomedin (OLF) homology domain may also perturb biosynthetic pathways and cellular homeostatic functions. Our recent studies have shown the OLF domain corresponds to a major translocational 'pause', an area where critical processes for normal TIGR biogenesis are expected to take place. Observations that Glu323Lys (and other mutations early in the OLF domain) altered the pattern of paused protein intermediates provide possible clues to previously unexplained pathogenetic mechanisms. HTM cell transfection studies using TIGR green fluorescent protein (GFP) fusions showed increased and altered distribution of the expressed protein with constructs missing the OLF domain, an effect also found with the Pro370 Leu mutation for early-onset glaucoma. The data suggest an activation of stress/apoptotic pathways in HTM cells as a potential mechanism for environmental/genetic interactions in glaucoma pathogenesis. PMID- 11026981 TI - Glaucoma medical treatment: philosophy, principles and practice. AB - There have been numerous recent advances in the management of glaucoma, not least the development of new drugs to help manage raised intraocular pressure. In addition, the concepts of improving blood flow to the optic nerve head and neuroprotection are currently provoking considerable interest. This article considers the aims and philosophy of glaucoma drug therapy, summarises some of the basic facts and principles of modern glaucoma medications, and suggests a practical approach to the choice of therapy. PMID- 11026982 TI - Perspectives in trabecular surgery. AB - The aim of trabecular surgery is to selectively combat the diseased structure central to the pathogenesis of chronic open-angle glaucoma, thereby reducing potential hazards during and after conventional filtering procedures. This overview considers new techniques in ab interno trabecular surgery. Special emphasis is placed on the description of each novel technique, its instrumentation, presumed mechanism of action and clinical results. Trabecular aspiration is evaluated as a method of clearing intertrabecular spaces of extracellular debris in pseudoexfoliation glaucoma with or without simultaneous cataract surgery or goniocurettage, while laser trabecular ablation is discussed for the treatment of absolute glaucomas. Where corneal haze has formed visualisation of the anterior chamber angle structures and trabecular surgery is performed with the aid of a microendoscope. Although the results are very promising it should be understood that some of these procedures are still in the experimental phase and are undergoing careful clinical evaluation, leaving plenty of room for refinements and further developments. PMID- 11026983 TI - Sinusotomy and deep sclerectomy. AB - Non-penetrating filtering surgery (NPFS) started in 1962 with the first sinusotomy performed by Kraznov. At that time, the author of this new technique believed that the outflow resistance in the majority of cases of primary open angle glaucoma was located at the level of scleral aqueous drainage veins and not in the trabeculum. He therefore developed a safe NPFS technique, leaving in place the trabeculum and the inner wall of Schlemm's canal. Because of difficulties with the microsurgical technique and the small reduction in intraocular pressure (IOP) compared with trabeculectomy, sinusotomy was abandoned. In the last decade, with the widespread use of operating microscopes, NPFS has been the subject of renewed interest. IOP reduction with the new NPFS techniques is comparable to that obtained with trabeculectomy, with significantly lower pre- and post operative complications. The new NPFS techniques such as deep sclerectomy, ab externo trabeculectomy and viscocanalostomy present definitively different mechanisms of filtration compared with early sinusotomy. This article will review the history of NPFS as well as describing the different new non-penetrating filtering surgeries. PMID- 11026984 TI - Modulating conjunctival wound healing. AB - Advances in molecular and cell biology have led to an expansion in our knowledge and understanding of the processes involved in wound healing. We review existing and potential therapies modulating the conjunctival scarring response, with particular reference to glaucoma filtration surgery. We discuss how the refinement of present antimetabolite regimens can minimise complications and improve surgical results, and advocate their use in carefully selected patient groups. Perhaps the most promising approach is targeting biological molecules. Hence, use of fully human neutralising monoclonal antibodies to the growth factor TGF beta has potential as a useful strategy for modifying conjunctival scarring. Combination therapies may also afford an improved therapeutic index. It is hoped that future therapies can offer safer, more specific, focal and titratable treatment, with far-reaching clinical applications. PMID- 11026985 TI - CJD and the eye. PMID- 11026986 TI - Pupil dilation and driving. PMID- 11026987 TI - Creutzfeldt-Jakob disease and the eye. I. Background and patient management. AB - This article attempts to summarise our current understanding of TSEs as they affect man. Specific aspects relevant to ophthalmological practice, in particular the management of patients in day-to-day clinical practice and with respect to corneal transplantation, have been discussed. In the companion article we discuss the specific ophthalmic and neuro-ophthalmic features of these diseases. PMID- 11026988 TI - Creutzfeldt-Jakob disease and the eye. II. Ophthalmic and neuro-ophthalmic features. PMID- 11026990 TI - Surgery for full-thickness macular holes with short-duration prone posturing: results of a pilot study. AB - PURPOSE: To see whether surgical success and complication rates in surgery for full-thickness macular holes (FTMH) followed by 5 days prone posturing are comparable to those obtained with longer posturing regimes recorded in the literature. METHODS: A pilot study was carried out of pars plana vitrectomy, autologous platelet adjunct and 16% C2F6 tamponade followed by 5 days prone posturing in 38 eyes of 34 patients with idiopathic FTMH. A follow-up postal questionnaire was used to assess patients' perception of posturing and outcome. RESULTS: Fifty-three per cent of eyes gained 2 or more lines of Snellen acuity. Twenty-four per cent of patients with symptom duration of 12 months or less (29 patients) achieved a visual acuity of 6/12. Fifty-eight per cent of patients achieved N8 or better near vision. The only significant predictor of post operative Snellen acuity was the stage of the hole (p = 0.02). Eighty-six per cent of questionnaire respondents felt that surgery had improved their quality of life. Eighty-seven per cent of all patients reported a reduction in, or elimination of, metamorphopsia. Fifty-four per cent of patients described posturing for 5 days as difficult or very difficult. Five patients admitted to posturing for less than 12 h a day, but all stated that they had postured for the full 5 days. Cataract was the commonest complication observed in this series (42% of patients have had or been listed for cataract surgery). CONCLUSIONS: Five days of prone posturing following vitrectomy for FTMH with autologous plaletet concentrate and C2F6 tamponade afforded success and complication rates comparable to those in published studies with longer posturing times. PMID- 11026989 TI - The effect of pupil dilation with tropicamide on vision and driving simulator performance. AB - PURPOSE: To assess the effect of pupil dilation on vision and driving ability. METHODS: A series of tests on various parameters of visual function and driving simulator performance were performed on 12 healthy drivers, before and after pupil dilation using guttae tropicamide 1%. A driving simulator (Transport Research Laboratory) was used to measure reaction time (RT), speed maintenance and steering accuracy. Tests of basic visual function included high- and low contrast visual acuity (HCVA and LCVA), Pelli-Robson contrast threshold (CT) and Goldmann perimetry (FIELDS). Useful Field of View (UFOV--a test of visual attention) was also undertaken. The mean differences in the pre- and post dilatation measurements were tested for statistical significance at the 95% level using one-tail paired t-tests. RESULTS: Pupillary dilation resulted in a statistically significant deterioration in CT and HCVA only. Five of 12 drivers also exhibited deterioration in LCVA, CT and RT. Little evidence emerged for deterioration in FIELDS and UFOV. Also, 7 of 12 drivers appeared to adjust their driving behaviour by reducing their speed on the driving simulator, leading to improved steering accuracy. CONCLUSIONS: Pupillary dilation may lead to a decrease in vision and daylight driving performance in young people. A larger study, including a broader spectrum of subjects, is warranted before guidelines can be recommended. PMID- 11026991 TI - Long-term follow-up of ischaemic retinopathy in the antiphospholipid syndrome with lupus-like disease. AB - PURPOSE: Antiphospholipid syndrome (APS), as an acquired prothrombotic disorder, is increasingly being recognised as an important cause of systemic venous and arterial thrombosis. The defining feature of the condition is the presence of raised levels of antibodies to negatively charged phospholipids in the serum. METHODS: We describe 2 cases of APS with ocular involvement and review the recent literature. Both patients experienced acute visual loss. It was the presenting symptom in one case--a finding that led to the diagnosis of the syndrome. RESULTS: Management with anticoagulation therapy, in which the International Normalised Ratio (INR) has been maintained at or above 3, resulted in reperfusion of the ischaemic retina and stabilisation of the retinopathy in one patient, whilst in the other case, where the INR was less than 3, irreversible visual loss occurred. CONCLUSION: Anticoagulation with warfarin appears to result in reperfusion of ischaemic retina with stabilisation of the neovascular process when the INR is greater than 3. PMID- 11026992 TI - Clinical evaluation of 'local contrast enhancement' for oral fluorescein angiograms. AB - PURPOSE: To describe and assess the clinical impact of 'local contrast enhancement' on oral fluorescein angiograms. METHODS: Oral fluorescein angiograms (OFA) were performed on 12 patients with a range of diabetic retinopathy. The digital images were processed using two enhancement techniques: histogram equalization, a widely available method; and 'local contrast enhancement'. Twelve control images and 24 enhanced images were randomised and subjectively graded for clarity between 1 and 100 on a visual analogue scale by two masked observers. A score of 50% was chosen as the cut-off for clinically useful images. RESULTS: The mean score for the unenhanced images was 38.8% (SD 19.4); the histogram equalization performed better at 54.7% (SD 10.0) (p = 0.016) and 'local contrast enhancement' at 69.4% (SD 13.6) (p < 0.001). These results equate to a 14.7% improvement in comparison with histogram equalization and a 30.3% improvement with the 'local contrast enhancement' technique. All the 'local contrast enhancement' images were clinically useful, compared with 58.3% of the histogram equalization images (chi 2 2.08, p = 0.2), and 33.3% of the control images (chi 2 = 6.75, p = 0.01). CONCLUSION: This pilot study has demonstrated the power of 'local contrast enhancement' techniques for low-contrast fluorescein angiograms. Using 'local contrast enhancement' oral fluorescein angiograms can give high quality images and may allow safer patient investigation. PMID- 11026993 TI - Ocular fundus abnormalities in children born before 29 weeks of gestation: a population-based study. AB - PURPOSE: Preterm birth has been found to be associated with increased morbidity of the central nervous and vascular tissues. To investigate the influence of preterm birth on the optic disc and retinal vessels, we examined the ocular fundus in school-aged children born very preterm. METHODS: A prospective, population-based study was performed in 50 very preterm children (median age 7 years, range 5-9 years) with a median gestational age at birth of 27 weeks (range 24-28 weeks) and a median birth weight of 1055 g (range 450-1520 g). The ocular fundus was examined by ophthalmoscopy in 50 children, and the optic nerve and retinal vessel morphology was evaluated by digital image analysis of ocular fundus photographs in 45 of these children. RESULTS: The median optic disc area was significantly smaller (p = 0.0002) in the preterm children compared with a reference group. There was no difference in cup area and, consequently, the rim area was significantly smaller (p = 0.0002) in the preterm children. Children with early signs of brain lesions commonly had a rim area below the median of the reference group. Preterm children also commonly had an abnormal retinal vascular pattern that was independent of a previous history of retinopathy of prematurity. CONCLUSION: Very preterm birth was associated with subnormal optic disc and rim areas and an abnormal vascular pattern. The findings clearly demonstrate the effect of preterm birth on the development of these structures. The long term clinical prognosis of these findings has yet to be determined. PMID- 11026994 TI - In vitro fertilisation and stage 3 retinopathy of prematurity. AB - PURPOSE: To re-examine the risk of children born by assisted conception developing stage 3 retinopathy of prematurity (ROP) and to define whether the risk of ROP varies with the method of assisted conception. METHODS: This was a retrospective study carried out between December 1995 and December 1998 of infants in a single neonatal unit serving the Brent and Harrow area of North West Thames requiring screening and treatment of ROP. The infants screened were identified from the ROP screening database. Those conceived by in vitro fertilisation (IVF) and other forms of assisted conception were identified by reviewing the neonatal notes and the maternal obstetric records. Birth weight, gestational age and the type of assisted conception were recorded. The presence or absence of any stage of ROP, its location and severity and the cases requiring treatment were recorded. RESULTS: One hundred and seventy-nine infants fulfilled the screening criteria during this period. Acute ROP was detected in 32.4% (58 infants) and stage 3 ROP developed in 15.6% (28 infants). Twenty-one infants (11.7%) were born after assisted conception, with 12 (6.7%) being conceived by IVF. The others were conceived on clomiphene (8) or after intrauterine insemination (1). Assisted conception accounted for 21.4% of all those reaching stage 3 disease and 28.6% of those infants requiring treatment. Of the 12 infants conceived by IVF, 41.6% (5 infants) developed acute ROP which progressed to threshold ROP in all infants (100%). Of the assisted conception babies requiring treatment for ROP, 83.3% were conceived by IVF. The other child had been conceived on clomiphene. The gestational age and birth weight of the IVF infants reaching stage 3 ROP were 26.6 +/- 0.89 weeks and 937 +/- 170.2 g. The gestational age and birth weight in the rest of the infants reaching stage 3 ROP were lower than in those conceived by assisted conception (25.739 +/- 1.13 weeks and 735.29 +/- 117.70 g); however, this did not approach statistical significance (p = 0.35 and p = 0.13, respectively). CONCLUSIONS: In this study 11.7% of the group requiring screening were conceived by assisted conception. Of all babies requiring treatment for ROP, 28.6% were born after assisted conception. Of the assisted conception group, 83.3% were conceived by IVF. Assisted conception using IVF rather than other techniques appears to be the major risk factor for the development of threshold ROP. We would advise increased vigilance when screening babies conceived by the IVF methods of assisted conception. PMID- 11026995 TI - Vigabatrin-associated visual field defects in children. AB - PURPOSE: Vigabatrin (Sabril), a drug that blocks GABA transaminase, has been used in the treatment of epilepsy since 1989. There have been reports of irreversible constriction of the visual field in adult patients related to vigabatrin (VGB) therapy, resulting in reduced VGB usage in adults. Although used as a second or third line agent in adults, in children it is often considered as a first line treatment for several subgroups of seizures in spite of there being no way, in the majority of cases, to monitor visual fields. Some of these children have a pre-existing visual field defect as part of their primary disorder. We aimed to identify whether visual field loss due to VGB was occurring in our hospital. METHODS: We have studied the results of ophthalmic examination in 14 children on VGB at Great Ormond Street Hospital who were able to perform Goldmann visual fields. RESULTS: Ten of the 14 patients had constriction of their visual fields attributed to VGB. In addition there were 2 patients with suspicious visual field defects thought to be due to VGB. There was pre-existing visual pathway damage in 4 cases and in 2 of these optic disc pallor increased in association with constricted visual fields. However, the optic discs were normal in 7 patients in spite of visual field constriction. Visual acuity was generally normal in spite of gross visual field constriction. CONCLUSIONS: We believe that VGB should be used with great caution where there is pre-existing visual pathway damage. In other cases the benefits should be considered in relation to the risks, which include irreversible visual field damage. At present visual fields can only be monitored by perimetry, which is often not possible in children with epilepsy. PMID- 11026996 TI - Sulfasalazine in the prevention of anterior uveitis associated with ankylosing spondylitis. AB - PURPOSE: To assess the effects of sulfasalazine in preventing recurrences and reducing the severity of anterior uveitis associated with ankylosing spondylitis and chronic intestinal inflammation. METHODS: Twenty-two patients with anterior uveitis associated with ankylosing spondylitis were studied. Ten patients were randomised to receive oral sulfasalazine (group 1) and 12 patients randomised to no treatment (group 2); all were followed for 3 years. Blood-aqueous barrier permeability was determined by fluorophotometry and bowel biopsies were taken. RESULTS: A statistically significant difference was observed between the two groups regarding the number of recurrences of uveitis (p = 0.016). The blood aqueous barrier permeability was significantly higher during acute attacks in group 2 (group 1: 31.3 +/- 26.4 x 10(-4) min-1 vs group 2: 66.2 +/- 28.5 x 10(-4) min-1; p = 0.019) but not during the disease-free period. We observed a higher incidence of chronic intestinal inflammation at the end of the study in group 2 (group 1: 3/8 vs group 2: 7/9, p = 0.153). No relation was observed between blood aqueous barrier permeability and the number of recurrences. The number of patients with severe persistent posterior synechiae at the end of the study was higher in group 2 (group 1: 4 patients before and 4 patients at the end; group 2: 4 patients before and 8 patients at the end; p = 0.65). CONCLUSION: Sulfasalazine may be beneficial in preventing recurrences and reducing the severity of anterior uveitis associated with ankylosing spondylitis. PMID- 11026997 TI - Rifabutin-induced uveitis with inflammatory vitreous infiltrate. AB - PURPOSE: To describe an ocular complication of rifabutin therapy in the treatment of Mycobacterium avium complex (MAC) infection in the absence of HIV infection or AIDS. METHODS: Three patients on rifabutin therapy for MAC chest infection developed anterior uveitis with vitreous infiltrates. The clinical course and visual outcome are discussed. RESULTS: The ocular signs of anterior segment inflammation along with vitreous infiltrate resolved on topical steroid therapy without long-term visual sequelae. CONCLUSION: This report highlights the potential of uveitis in patients treated for MAC infection with rifabutin without concurrent HIV infection or AIDS. PMID- 11026998 TI - Riolan's muscle: action and indications for botulinum toxin injection. AB - PURPOSE: To study the effect of injecting botulinum toxin into the region of Riolan's muscle in three conditions, namely the typical form of essential blepharospasm, the palpebral form of essential blepharospasm and hemifacial spasm. METHOD: Six patients with the typical form of essential blepharospasm and 4 patients with the palpebral form of essential blepharospasm, all of whom had previously been treated with conventional bilateral periorbital injections, were treated with injections of the toxin into the region of Riolan's muscle at the medial and lateral extremities of the upper lids. Thirty patients with hemifacial spasm, all of whom had previously been treated with conventional periorbital injections, were treated with injections of the toxin into the region of Riolan's muscle at the medial and lateral extremities of the upper lid on the affected side. RESULTS: Five of 6 patients with typical essential blepharospasm preferred the Riolan's injections and one had no preference. All of the four patients with the palpebral form of essential blepharospasm preferred the Riolan's injections. They, previously, had hardly been able to open their eyes. Twenty-six of the patients with hemifacial spasm preferred the Riolan's injections; the other 4 decided to continue with periorbital injections. The amount of toxin used in this new method of treatment is considerably less than that used in conventional methods for these diseases. CONCLUSION: Riolan's injections of botulinum toxin are the preferred modality of treatment for all types of blepharospasm and cost considerably less. PMID- 11026999 TI - The House-Brackmann system and assessment of corneal risk in facial nerve palsy. AB - PURPOSE: The House-Brackmann (HB) facial nerve grading system is widely used by ENT/head and neck surgeons, but is perhaps of less value to the ophthalmologist. Our aim was to assess the value of the numeric portion of this system in identifying those patients with facial nerve palsy who are at risk of corneal complications. We also sought to identify other factors that might be predictive of such complications. METHODS: Forty-two patients (43 palsies) were studied prospectively. The HB grade was recorded together with measurements of levator function, upper lid closure, Bell's phenomenon, lagophthalmos, ectropion, lower lid retraction and corneal sensation. Conjunctival injection and corneal staining were also graded. ROC (receiver operating characteristic) curves were used to assess the value of each parameter as a screening test for corneal complications. RESULTS: There was no cut-off of HB grade, levator function, Bell's phenomenon, ectropion or lower lid retraction that was sufficiently sensitive and specific to screen for corneal complications. However, on assessing lagophthalmos and upper lid closure, cut-offs with more favourable sensitivities and specificities were identified. CONCLUSIONS: The numeric portion of the HB grading system is not a useful guide in identifying those patients with facial nerve palsy who are at risk of corneal complications. Measurements of lagophthalmos and upper lid closure, alone or in combination with other tests, may be of more value. PMID- 11027000 TI - Diagrammatic representation of lacrimal disease. AB - PURPOSE: To design a diagrammatic record of lacrimal disease using easily interpretable symbols. METHODS: This diagrammatic record was developed from existing models. A simple outline for the eyelids, lacrimal drainage apparatus and nose is used. The periocular, lid, dye test, syringing and nasal findings are represented by standard symbols. RESULTS: Examples illustrating a range of lacrimal findings, before and after surgery, are shown. CONCLUSIONS: A diagrammatic record of lacrimal findings which can be easily understood offers a sound basis for planning and monitoring lacrimal surgery. It is not necessary to memorize each diagram, only the principles on which they are drawn. PMID- 11027001 TI - Is the first post-operative day review necessary following uncomplicated phacoemulsification surgery? AB - PURPOSE: To assess the necessity for first post-operative day review in determining the need for post-operative intervention in patients who had uncomplicated phacoemulsification surgery. METHODS: A retrospective study was carried out to review the first post-operative day findings in patients who underwent uncomplicated phacoemulsification surgery by a single surgeon between January 1997 and March 1998. The findings analysed were wound integrity, corneal clarity, anterior chamber activity, intraocular pressure and the intraocular lens status. The need for medical or surgical intervention was also analysed. Those eyes that had coexisting ocular pathology such as glaucoma, ocular hypertension, uveitis, trauma or previous intraocular surgery were excluded from the study. Fisher's exact test was used to compare the difference between the groups. RESULTS: Seventy-one eyes of 71 patients who underwent an uncomplicated phacoemulsification procedure were included in the study. Intraocular pressure of 30 mmHg or greater was found in 7 eyes (10%), all of which also had corneal oedema. These patients received acetazolamide SR 250 mg twice daily for 3 days. Another 21 eyes (30%) had corneal oedema for which no specific treatment was given. The intraocular pressure had returned to baseline and corneal oedema resolved by the first clinic follow-up in 1-2 weeks. None of the 71 patients needed surgical intervention in the post-operative period. CONCLUSION: First post operative day review is necessary as it gives an opportunity to manage the post operative rise in intraocular pressure. PMID- 11027002 TI - Use of the GDx to detect differences in retinal nerve fibre layer thickness between normal, ocular hypertensive and early glaucomatous eyes. AB - PURPOSE: The GDx is a scanning laser polarimeter that has been developed to allow the quantitative analysis of retinal nerve fibre layer (RNFL) thickness. The purpose of this study was to determine whether differences in the RNFL thickness between normal, ocular hypertensive and glaucomatous eyes could be detected using the GDx. METHODS: RNFL analysis was carried out using the GDx on 33 normal, 145 ocular hypertensive (OHT) and 44 glaucomatous (POAG) eyes. The inclusion criteria for entry into the study did not include the clinical appearance of the RNFL or optic disc, thus eliminating an important source of selection bias. The Kruskal Wallis one-way analysis of variance was used to determine whether any significant differences existed among the groups in mean total and quadrantic RNFL thickness. When significant differences were found, specific inter-group analysis was carried out using the Mann-Whitney U-test. RESULTS: Significant differences in RNFL thickness were found for the mean total, superior and inferior quadrant values between normals and OHT eyes as compared with POAG eyes. No significant differences were found for these values between the normal and OHT eyes. CONCLUSION: Analysis of RNFL thickness using the GDx was able to detect differences between POAG eyes compared with normal and OHT eyes, although there was considerable overlap between groups. Further assessment of the technique is required to determine whether it may be useful in screening for the presence of early glaucomatous damage. PMID- 11027003 TI - Nerve fibre layer measurement of the Hong Kong Chinese population by scanning laser polarimetry. AB - PURPOSE: To obtain normal retinal nerve fibre layer (RNFL) measurements by scanning laser polarimetry in a local Hong Kong Chinese population and to identify the correlation of the measurements with age. METHODS: One hundred and fifty-nine normal Hong Kong Chinese volunteers of different ages were recruited for this study. RNFL values were measured using a Nerve Fibre Analyzer GDx (Laser Diagnostic Technologies, CA). RESULTS: Mean peripapillary RNFL measurements at the superior, inferior, temporal and nasal regions were 95.0 +/- 15.6, 97.0 +/- 16.3, 47.0 +/- 9.0 and 43.9 +/- 15.1 microns respectively (mean +/- SD). There was a highly significant negative correlation in average RNFL values with increasing age (approximately 1.9 microns per decade, p = 0.001). A significant negative correlation with increasing age was also identified in the RNFL values in all four regions and they decreased by similar amounts (p < 0.05). No correlation with age was observed for relative ratio parameters such as superior/nasal ratio, inferior/nasal ratio and max. modulation (p > 0.05). CONCLUSION: RNFL values of the Hong Kong Chinese population appeared to decrease over time cross-sectionally. Since they decreased by similar amounts at four regions, use of relative ratio parameters for analysis has an advantage over absolute values, as they were not affected by age. PMID- 11027005 TI - Lower lip trauma: an unusual side-effect of peribulbar anaesthesia. PMID- 11027004 TI - Synthesis pattern of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) in human explant organ cultures after treatment with latanoprost and dexamethasone. AB - PURPOSE: To determine changes in production of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the ciliary body, the trabecular meshwork and the retinal pigment epithelium induced by both prostaglandins and corticosteroids. METHODS: Explant organ cultures were removed by a scleral incision 3 mm posterior to the limbus. Retinal pigment epithelium was grown to confluence. Organ and cell cultures were treated with latanoprost and/or dexamethasone for 72 h. The activity of MMP-2, -3 and -9 was assessed using zymography. The synthesis pattern of MMPs and TIMP-1 and -2 was identified using immunostaining. RESULTS: Treatment of explant organ cultures with 10 micrograms/ml of latanoprost induced a mean upregulation of MMP-2 by 36%, MMP-3 by 112% and MMP-9 by 156% as seen by zymography. Dexamethasone 500 nm reduced the amounts of secreted MMP-2 by 13%, MMP-3 by 69%. MMP-9 was not detectable in the media of corticosteroid-treated explant organ cultures. The addition of 10 micrograms/ml of latanoprost to dexamethasone-treated cultures increased MMP-2 by 14%, MMP-3 by 43% and MMP-9 by 49%. Using immunohistochemistry we found staining with antibodies against MMP-2, -3, -9 and TIMP-1 and -2 within the ciliary body, and only to a lesser degree in the trabecular meshwork. Latanoprost treatment caused an increase of 29% in MMP-2 (p < 0.0001), 98% in MMP-3 (p < 0.0001) and 108% in MMP-9 (p < 0.0001). Dexamethasone reduced the staining for MMP-2 by 32% (p < 0.0001), for MMP-3 by 33% (p < 0.0001) and for MMP-9 by 83% (p < 0.0001). Almost no change in staining for MMPs was detectable in the trabecular meshwork. Neither latanoprost treatment nor dexamethasone induced significant changes (p < 0.93) in the secretion of TIMPs. In the media of non-treated retinal pigment epithelium (RPE) cells the only MMP detected was MMP-2. RPE cells in culture did not respond to either treatment with a change in their MMP secretion. CONCLUSION: We detected a profound upregulation of both MMP-3 and MMP-9 and a mild induction of MMP-2 through latanoprost in the ciliary body, but not the trabecular meshwork or RPE cells. Corticosteroids, on the other hand, downregulated MMP expression in both tissues. This inhibiting effect of corticosteroids on MMP production was reversed by latanoprost. PMID- 11027006 TI - Primary congenital upper eyelid entropion in association with Duane's retraction syndrome. PMID- 11027007 TI - Necrosis of the lid due to Pseudomonas aeruginosa. PMID- 11027008 TI - Ptosis repair in a patient with oculopharyngeal dystrophy: brow suspension using autogenous fascia lata by spinal anaesthesia. PMID- 11027009 TI - Orbital involvement with angiolymphoid hyperplasia with eosinophilia--a benign condition for the practising ophthalmologist to be aware of. PMID- 11027010 TI - Bilateral multifocal choroidal metastases as the first manifestation of a breast carcinoma. PMID- 11027011 TI - Clear cell thyroid carcinoma metastatic to choroid: clinicopathological study of a case. PMID- 11027012 TI - Unilateral retinitis pigmentosa in a woman and pigmented paravenous chorioretinal atrophy in her daughter and son. PMID- 11027013 TI - Traumatic hyphaema and sickle cell retinopathy in a patient with sickle cell haemoglobin E (HbSE) disease. PMID- 11027014 TI - Leukaemia manifesting as uncontrollable proliferative retinopathy in a diabetic. PMID- 11027016 TI - Worsening of an existing hemispheric retinal vein occlusion following further hypotony procedure. PMID- 11027015 TI - Drug or drusen? Central retinal vein occlusion in a young healthy woman with disc drusen. PMID- 11027017 TI - Glaucoma mimicked by carotid-ophthalmic artery aneurysm: a rare insidious cause of progressive visual loss. PMID- 11027018 TI - Acute angle closure glaucoma presenting in a young patient after administration of paroxetine. PMID- 11027019 TI - Improved visual acuity in an eye with glaucomatous optic atrophy following loss of the good eye. PMID- 11027020 TI - Tono-Pen tonometer and corneal thickness. PMID- 11027021 TI - Capsular glaucoma. PMID- 11027022 TI - Surgically induced scleral necrosis. PMID- 11027023 TI - Peroperative retinoscopy as a predictor of final post-operative refraction. PMID- 11027024 TI - Acute angle closure glaucoma: an evaluation of a protocol for acute treatment. PMID- 11027025 TI - The management of acute angle-closure glaucoma. PMID- 11027026 TI - An improved method for the measurement of added vegetable fats in chocolate. AB - A method for identifying refined vegetable fats added to chocolate (cocoa butter equivalents, CBEs) was combined with established quantitative methods for determining the level of vegetable fat added to cocoa butter with the aim of providing improved precision. The identification of fats was based on the analysis of sterol and triterpene alcohol degradation products formed during the processing of the fat. The procedure was able to successfully discriminate between 95% of pairs of fats from a set (33) of CBE-type vegetable fats. Subsequent analysis of 80 mixtures of four CBEs with chocolate successfully identified, on cross-validation, 94% of the samples. Combining the qualitative procedure with established quantitative methodology, based on the analysis of triacylglycerols, improved the method precision from +/- 2.1% to +/- 0.3% (5% addition of CBE at 95% confidence). Identifying the fat analytically permits the use of quantitative methods for determining the level of added fat in chocolate that have improved precision in comparison with the measurement of an unidentified fat. This may obviate the need to use factory inspection as a means to identify the ingredients of a product and monitor compliance with proposed legislation. PMID- 11027027 TI - Analysis of raw materials, intermediates and subsidiary colours in Food Yellow No. 5 (Sunset Yellow FCF) by LC/MS. AB - Raw materials, intermediates and subsidiary colours in Food Yellow No. 5 (Sunset Yellow FCF) were determined using liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization. A gradient consisting of acetonitrile and 0.04% aqueous ammonium carbonate solution was used for the HPLC mobile phase. Quasi molecular ions of impurities were used as monitor ions. It was necessary to use fragment ions of the sodium salts of 6-hydroxy-5-phenylazo-2-naphthalenesulphonic acid (SS-AN) and 4-(2-hydroxy-1-naphthylazo) benzenesulphonic acid (2N-SA) as monitor ions because the compounds are not resolved by chromatography and have the same molecular weight. Fifteen samples of commercial Sunset Yellow FCF were examined. The results obtained by UV-Vis spectroscopy were in good agreement with the results of LC/MS analyses. The detection limits of the impurities in Sunset Yellow FCF ranged from 0.01 to 0.1%. PMID- 11027028 TI - The effect of storage at 4 degrees C on the stability of ampicillin residues in raw milk. AB - Raw milk samples collected from tanker trucks are routinely screened for beta lactam antibiotic drug residues using rapid screening tests. If drug residues are detected, the milk may be shipped on ice blocks to a laboratory for further analysis. A study was conducted to determine the stability of ampicillin in raw milk stored at +4 degrees C in order to predict if shipping the milk would result in the degradation of ampicillin residues. Milk samples were spiked with 20 ppb ampicillin, stored at +4 degrees C and -70 degrees C for 1-6 days, and then analysed by HPLC with fluorescence detection. No loss of ampicillin residues was found in milk stored at +4 degrees C for 1-6 days. PMID- 11027029 TI - Cadmium in organs and tissues of horses slaughtered in Italy. AB - The cadmium content of muscle, liver, kidney and blood samples from 62 horses slaughtered in Italy was investigated. Cadmium was determined by graphite furnace atomic absorption spectrometry (GFAAS) after wet digestion of the samples. The mean and median contents of all samples were (on a fresh weight basis) 75 and 41 micrograms kg-1 for muscle, 2.46 and 2.10 mg kg-1 for liver, 20.0 and 13.5 mg kg 1 for kidney. The cadmium level in blood samples was always below 6 micrograms l 1. The cadmium concentrations in muscle, liver and kidney were found to be related to the life span of the specimens and increased with age. Females exhibited higher levels than males, but this difference was significant only in the case of muscle tissue. The geographical origin was recognized as the main factor influencing the cadmium content of the equine specimens analysed. The differences between horses coming from the three main breeding countries considered in this study (Poland, Lithuania, Hungary) were marked and statistically significant. The average intake of cadmium from equine meat was estimated for the general population and for population groups resident in areas with high consumption of this food item. While the cadmium intake from equine meat for the average consumer accounts for about 1% of the total cadmium intake estimated for Italy, in the latter case the enhanced consumption of equine muscle is often accompanied with the consumption of substantial quantities of equine liver and this may lead to high cadmium intakes. PMID- 11027030 TI - Pre-harvest accumulation of deoxynivalenol in sweet corn ears inoculated with Fusarium graminearum. AB - Three types of commercial sweet corn hybrids [surgary (su1), shrunken or 'supersweet' (sh2) and surgary enhancer (se1)] were silk channel inoculated in 1996 and 1997 with a macroconidial suspension of Fusarium graminearum to determine how early the mycotoxin deoxynivalenol accumulates in kernels. Disease symptoms rapidly developed on all hybrids and were apparent 4 days after inoculation. Symptoms stabilized by 28 days after inoculation. Toxin levels were greater than 1 microgram/g in kernels as early as 2 weeks after silk emergence and rapidly increased to extremely high levels. Susceptibility in all hybrids decreased as the silk dried out. Deoxynivalenol concentrations were correlated to disease severity. There was some indication that the sh2 genotype was more susceptible than the su1 or se1 genotypes. These results suggest that improvement needs to be made in sweet corn with respect to resistance to gibberella ear rot. PMID- 11027031 TI - Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants. AB - The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 microns-thick oriented PP) into the food stimulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials. PMID- 11027032 TI - Influence of trace metals, acids and ethanol in food-simulating liquids on the retention of alpha-tocopherol in low-density polyethylene film. AB - The retention of alpha-tocopherol (vitamin E) in low-density polyethylene (LDPE) film was investigated when in contact with different food-simulating liquids. Variations of the aqueous food simulants approved by the EU for testing of interactions between packaging materials and food, and an alternative fatty food simulant (95% aqueous ethanol) were used to study the influence of trace metals and acids, as well as variation in pH and ethanol concentration, on the stability of alpha-tocopherol in the LDPE film. The presence of trace metals in aqueous media accelerated loss of alpha-tocopherol from the film, while the addition of citric or ascorbic acid counteracted this loss. In contact with aqueous ethanol at a concentration of 50% or higher, the loss of alpha-tocopherol from the film was total. However, with solutions containing less than 50% ethanol, the antioxidant was almost completely retained in the film, unless the temperature was increased from 6 to 40 degrees C. For contact with solutions containing ethanol or trace metals, an increase in temperature resulted in a greater loss of alpha-tocopherol from the LDPE film. PMID- 11027033 TI - Nutritional status of Kuwaiti elementary school children aged 6-10 years: comparison with the NCHS/CDC reference population. AB - The objective of this cross-sectional sampling survey was to assess the nutritional status of Kuwaiti elementary school children aged 6-10 years in comparison with the NCHS/CDC American reference population. The study comprised a multistage stratified random sample of 8957 children (4401 males and 4556 females) which amounts to about 12% of the target population. Weights and heights of children were measured. Obesity was defined as weight-for-height (W/H) 2.00 to 5.00 standard deviation (SD) scores of the reference population. Underweight and short stature were defined as weight-for-height (W/H) and height-for-age (H/A) 4.00 to -2.00 and -5.00 to -2.00 SD scores, respectively. The results of this study showed that obesity was found to be significantly (P < 0.01) higher in males (15.7%) than in females (13.8%). Underweight was found to be almost equally prevalent among both genders (3.7% in males and 3.8% in females). Short stature (H/A) -5.00 to -2.00 SD scores) was found to be 7.1% (9.5% in males and 4.6% in females). When compared with an earlier study, changes in W/H were significant across most of the centile categories. However, significant changes in H/A were noted with less frequency than W/H; mostly in the upper centiles for both genders. More significant changes, however, were noted among males than females. In conclusion, Kuwaiti children were found to be heavier and shorter than American children. Moreover, the children were found to be fatter and slightly taller but nonetheless more nutritionally sound than a decade earlier. PMID- 11027034 TI - Age differences in physique of adult males aged 30 to 86 years in Rarotonga, the Cook Islands. AB - In the Pacific Region, some adult populations have shown a steady rise in overweight and obesity across the 1970s and into the 1990s. While younger adults have been shown to have lower body mass index (BMI) than older ones in both the least and most modernised Samoan populations, among intermediately modernised Samoan populations, BMI has been found to be higher in younger adults than in older ones. A survey in the Cook Islands carried out in 1966 showed no age group differences in height, weight and BMI among adult males, but significantly higher mean weight and BMI among adult males aged 30 years and above. The present analysis gives mean height, weight, BMI and skinfolds of adult males above 30 years of age on Rarotonga in 1996, and examines whether the BMI-age group relationship now shows a similar transitional pattern to that observed on American Samoa. In addition, the 1996 values are compared with values obtained in 1986, to determine whether changes in physique have taken place across this time. In the 1996 volunteer sample of 142 male Cook Islanders, older adults are significantly shorter, lighter, with lower BMI than younger adults. Furthermore, the younger adults of the 1996 survey are significantly heavier, with greater BMI than the 1986 sample. This suggests that the adult male Rarotongan population is in an intermediate position with respect to lifestyle transition, the secular trend in body size and increasing prevalence of obesity, and that there has been a rapid increase in body fatness prevalence among younger adults. PMID- 11027035 TI - Food safety knowledge and practices of streetfood vendors in a Philippines university campus. AB - A survey on food safety knowledge and practices of streetfood vendors from a representative urban university campus in Quezon City, Philippines was done. A face-to-face interview was conducted using a standardized survey tool containing 70 questions, which included queries on demographics and food safety knowledge and practices of streetfood vendors. Topics on food safety assessment in both practices and knowledge included: health and personal hygiene, good manufacturing procedures, food contamination, waste management, and food legislation. The study found that among the 54 streetfood vendors surveyed, knowledge on food safety concepts was established particularly on topics that dealt with health and personal hygiene, food contamination and good manufacturing procedures. However, vendors were shown to be not too knowledgeable in terms of food legislation and waste management. A significant gap between knowledge and practice on these topics was established and it was primarily attributed to the tendencies of street food vendors to compromise food safety for financial issues. Confusion in food legislation was established in this test microcosm because the purveyor of food safety regulations was not the local government health unit but the business concession office of the campus administration. The provision of continuous food safety education, some financial assistance through social services affiliations, and basic water and waste management utilities were recommended to diminish the gap between knowledge and practices of safe streetfood vending in school campuses. PMID- 11027036 TI - Variability of micronutrient content in enriched dairy and fruit products. Executive Group for Development of Nutrient Dense Foods for Frail Elderly. AB - Accurate measures of micronutrient levels in newly developed enriched foods are indispensable for valid labelling, for determination of ultimate levels for possible claimed health effects and for safety. To date, only a limited amount of public literature is available on the reproducibility of laboratory analysis and on the similarity within novel fortified or enriched foods. Within the framework of an intervention trial in frail elderly, the micronutrient content of eight different types of enriched foods were repeatedly examined. The variance in concentrations of the vitamins B1, B2, B6, folic acid, B12, C, D, and E was studied, in addition to levels of the minerals zinc, iodine, calcium, iron and magnesium. Four main factors, possibly contributing to the variability of these micronutrient levels, were investigated: (1) type of product, (2) type of laboratory (between-laboratory) reproducibility), (3) time of the year (month) in which analyses were carried out (May-October) and (4) freshness of the product. The type of product and laboratory emerged as factors contributing mostly to the total variability in concentrations of vitamins (on average approximately 50% explained). However, none of the products consistently contained higher or lower levels compared to the other products and most target levels were met except for vitamin B2 in both dairy and fruit products and for folic acid and vitamin C in some fruit products. Differences between products were regarded as acceptable. Extensive evaluation of multiple lab results should be emphasised within the manufacturing process of enriched foods. PMID- 11027037 TI - Comparative study of two techniques for assessing fat intake in a Mediterranean diet. The influence of culinary oils changes. AB - The diet usually consumed by a community of 18 normolipemic monks was studied. It included olive oil as cooking fat (basal period BP). This diet was modified by substituting olive oil for sunflower oil during a 21-day period (experimental period EP). The precise individual weighing technique was utilised for the determination of intake, and by means of the food composition tables (FCTs) the energy and nutrient of the diet were calculated. By chemical analysis of the duplicate sample (CA) of the cooked dishes the lipid intake was also determined. Total serum cholesterol HDL and LDL cholesterol, serum triglycerides, AI and B 100 apolipoproteins and Lp(a) were also analysed. Similarly the absolute and relative values for SFA, MUFA and PUFA obtained by both techniques were also different and these were maintained when the (PUFA + MUFA)/SFA ratio was calculated by both techniques and in the two periods. The correction of the lipid intake by means of the CA technique did not improve either the correlation between the intake or the plasma lipid concentration or the cholesterolemia prediction (r = 0.83 for FCT and CA). PMID- 11027038 TI - Evaluation of the nutritional quality and acceptability of sorghum-based tempe as potential weaning foods in Tanzania. AB - Six types of sorghum (Sorghum bicolor)-based tempe were developed by blending with various combinations and quantities of either bambara groundnut, sesame, cowpea, pigeonpea, common bean, mungbean, soybean, or groundnut (peanut) seeds. Their mixtures were then soaked and solid-state fermented by Rhizopus oligosporus to produce tempe. Soaking resulted in the loss of minerals. Mould fermentation did not change the proximate and mineral composition, but increased the in vitro protein digestibility by 10 to 14.1%. Tempe processing reduced the tannin content by 30.4 to 94.0%. The protein content of tempe ranged from 16.0 to 18.6%; the amino acid score, 63.0 to 69.0%; protein energy, 14.2 to 16.8%; net dietary protein energy, 7.3 to 8.4%; and total energy, 416.0 to 441.0 Kcal/100 g of tempe. The developed tempe had protein quality and energy recommended for weaning foods. The deep-fried snacks were mildly acceptable. More research work and promotion are proposed in order to improve the acceptability of sorghum-based tempe as potential weaning or supplementary foods in Tanzania. PMID- 11027039 TI - Nutritional and anti-nutritional composition of velvet bean: an under-utilized food legume in south India. AB - Four accessions of the under-utilized legume, velvet bean (Mucuna pruriens var. utilis (Wall. ex Wight) Bak. ex Burck), collected from three different locations of Western Ghats, South India were analysed for proximate composition, mineral profiles, the protein fractions, amino acid profiles of total seed protein, in vitro protein digestibility and certain anti-nutritional factors to determine their potential as an alternative source to alleviate protein-energy-malnutrition among the people of South India. The major findings of the study were as follows: crude protein ranged from 20.2-29.3%, crude lipid 6.3-7.4%, total dietary fibre 8.7-10.5%, ash 3.3-5.5% and carbohydrates 49.9-61.2%. The energy level of the seed (1562-1597 kJ 100 g-1 DM) was comparable with commonly consumed Indian pulses. Mineral profiles, viz. sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc and manganese ranged from 43.1-150.1, 778.1-1846.0, 393.4 717.7, 174.9-387.6, 98.4-592.1, 10.8-15.0, 0.9-2.2, 5.0-10.9, 3.9-4.3 mg 100(-1) seed flour, respectively. The data on seed protein fractions revealed that the globulins constitute the major bulk of the seed protein as in most legumes. Profiles of amino acids of total seed proteins detected in the present study revealed that they contain relatively higher levels of all essential amino acids except threonine, leucine and lysine in black-coloured seed coat accessions and phenylalanine and tyrosine in white-coloured seed coat accession compared with the FAO/WHO (1991) requirement pattern. The in vitro protein digestibility of the legumes under study ranged from 72.4-76.9%. Anti-nutritional substances like total free phenolics, tannins, L-DOPA, trypsin inhibitor activity and phytohaemagglutinating activity also were investigated. The detected anti nutritional factors probably have little nutritional significance if the beans are properly processed. PMID- 11027041 TI - A comparative study on the physical characteristics and cooking quality parameters of commercial brands of Basmati rice. AB - Randomly selected samples of three popular commercial brands of Basmati rice (designated as A, B and C) available in Mumbai city were studied with respect to their proximate composition and differences in their quality evaluated in terms of length by breadth and elongation ratio, differential length by breadth ratio, water uptake, loss of solids in cooking water, swelling index and volume expansion ratio. The l/b ratio and elongation ratio varied between 4.47 to 4.81 and 1.45 to 1.59 respectively. The differential length by breadth ratio showed a wide variation from 5.88 to 9.48 among the three brands studied. Brand A recorded the highest length by breadth and elongation ratio whereas brand C recorded the highest differential length by breadth ratio. The water uptake values and the loss of solids in the cooking gruel varied between 172 to 262 and 0.87 to 2.70 (g/100 g of rice) respectively, among the three commercial brands. The swelling index and the volume expansion ratio did not show much variation among the three brands ranging between 28 to 31 for the former and 2.00 to 2.21 for the latter. Based on the differential length by breadth ratio, water uptake and swelling index values, which are the most reliable quality indices; brand C was found to be the best followed by brands A and B. The study of the texture of the cooked rice grains indicated that brand C had the softest grains, which was in conformity with the findings in cooking quality characteristics. PMID- 11027040 TI - Nutritional evaluation of protein and starch of mature Canavalia gladiata seeds. AB - Evaluation of true digestibility (TD), biological value (BV) and net protein utilisation (NPU) of diets made with raw and processed seed flour from mature Canavalia gladiata seeds were carried out with male Sprague-Dawley rats. The weight gain of the rats fed with diets containing raw whole seed and raw cotyledon alone were significantly lower (P = 0.05) than that of the group fed with the reference diet. Processing the cotyledons (dry-autoclaved or roasted) significantly (P < or = 0.05) increased the weight gain of the rats when compared to the weight gain of rats fed diets prepared with raw seed flour. NPU of raw (whole seed 13.8; cotyledon 27.6) was significantly lower (P < or = 0.05) than the reference diet (79.5). The NPU of processed samples was also significantly lower (P < or = 0.05) when (dry-autoclaved 25.1; roasted 25.1) compared to the reference diet fed group (79.5). The BV of the processed samples (dry-autoclaved 31.1; roasted 37.7) was significantly lower (P < or = 0.05) than that of raw (53.6) cotyledon. In contrast TD increased (P < or = 0.05) with processing (dry autoclaved 80.9; roasted 65.9) when compared to raw cotyledon (51.4). In vitro protein digestibility for the raw whole seed and cotyledon was 71.7 and 70.1% respectively. In vitro starch digestibility of raw and processed cotyledon flour samples indicated dry-autoclaved sample to have the highest digestibility. PMID- 11027043 TI - The future of paediatric otolaryngology. PMID- 11027042 TI - Changes of specific activity of lipase and lipoxygenase during germination of wheat and barley. AB - Changes of lipase and lipoxygenase specific activities were monitored during wheat and barley germination. Statistically significant correlation (r2 = 0.8148, P < or = 0.02) was found between the specific activities of lipase and lipoxygenase during wheat germination, whereas such correlation did not occur during barley germination. PMID- 11027044 TI - Predominant role of the chorda tympani nerve in the maintenance of the taste pores: the influence of gustatory denervation in ear surgery. AB - The effect on the taste pores of denervation of the chorda tympani nerve in the middle-ear cavity was studied comparing confocal laser microscopy with lingual nerve resection. Taste pore cells were stained for actin with rhodamine phalloidin and positive fluorescence was observed as a ring shape at the transverse cross sections. Within three days after chorda tympani nerve resection the ring reaction disappeared, although the pore morphology remained intact as seen by scanning electron microscopy. On the other hand, lingual nerve resection did not induce such rapid disappearance of the ring reaction. These results suggest that the chorda tympani nerve plays a predominant role in the maintenance of actin filaments in taste pore cells. PMID- 11027045 TI - Telescopic myringotomy and tube application. AB - Otoendoscopy is a new technique in otological surgery. The traditional surgery for otitis media with effusion (OME) is myringotomy and tube insertion using an operating microscope. In 45 children and five adults presenting to our department with otitis media with effusion, rigid endoscopic myringotomy and grommet tube insertion were performed using 2.7 mm diameter and 0 degree and 30 degrees angle telescopes under general anaesthesia for children and local anaesthesia for adults. The use of rigid endoscopes provides a large field of view, that is of excellent resolution and fidelity of colour as well as giving good side views. It can be performed in the out-patient clinic with little inconvenience to the patient and minimal risks. PMID- 11027046 TI - Screening for vestibular schwannoma: magnetic resonance imaging findings and management. AB - We report the contrast-enhanced magnetic resonance imaging (MRIg) findings from a series of 1139 patients who underwent screening to exclude a diagnosis of vestibular schwannoma. An acoustico-facial nerve bundle tumour was found in 3.1 per cent of patients imaged and vestibular schwannoma incidence is estimated at 1.4 per 100,000 population per annum. MRIg showed an abnormality in 14 per cent of patients; about one third of the findings may have accounted for the presenting symptom(s). A small number of patients had unexpected pathology revealed that required onward referral for further active management. PMID- 11027047 TI - Vestibular schwannoma presenting as sudden deafness. AB - In this study, we analysed the clinical and audiological features of 24 vestibular schwannoma patients presenting with sudden deafness who visited our hospital within 10 days after its onset. The relationships between the prognosis of hearing loss and the clinical and audiological features including the pure tone average (PTA), the presence of dizziness and/or vertigo, the tumour size and the auditory brain stem response (ABR) findings were evaluated. Hearing recovery depended on the PTA and tumour size, but even a small tumour induced sudden deafness. Impaired hearing completely recovered in five our of 24 patients. ABR findings indicated that the aetiology of sudden deafness in the vestibular schwannoma patients might vary in each patient. PMID- 11027048 TI - Manual dexterity and successful hearing aid use. AB - Fine dexterity of the hand might be expected to correlate with the successful use of a hearing aid. In this study, the manual dexterity of 30 individuals between 65 and 85 years using a hearing aid was tested by the Purdue pegboard test and the result was compared to the benefit obtained from the use of a hearing aid assessed by a questionnaire. Of the 30 individuals included in this study 16 were wearing a 'behind the ear' type of hearing aid and the rest had an 'in the ear' type of hearing aid. A comparison is made between the two types of heariing aids. The results demonstrate a correlation between manual dexterity and successful use of a hearing aid when all the 30 individuals were considered as a single group. However, no such correlation was found for 'in the ear' group when they were analysed separately. PMID- 11027049 TI - 'A parent's kiss': evaluating an unusual method for removing nasal foreign bodies in children. AB - Nasal foreign bodies are a common problem in children. Various techniques have been described for removing the offending articles. Positive pressure techniques have long been described, and have many potential advantages, but are not yet in widespread use. The aim of this study was to evaluate the efficacy, safety and acceptability of a technique of mouth-to-mouth blowing. Of 19 children with intranasal foreign bodies, the technique was successful in 15 (79 per cent). The procedure caused little or no distress to the child, and no complications were encountered. All parents found the technique acceptable. This technique for nasal foreign body removal is, as far as we are aware, the first to be subjected to any prospective evaluation of effectiveness. We suggest that it should be used as the first line management for removing nasal foreign bodies in young children. PMID- 11027050 TI - Laser palatoplasty: evaluation of patient benefit using the Glasgow benefit inventory. AB - Short-term results of laser palatoplasty have been encouraging. This study was undertaken to evaluate if there is a deterioration of benefit with time as seen in uvulopalatopharyngoplasty (UPPP). The Glasgow benefit inventory (GBI) was applied retrospectively by means of a postal questionnaire to measure the quality of benefit and to evaluate any deterioration in benefit with increasing post operative time. Our results show that laser palatoplasty benefited 53 per cent of patients. With increasing post-operative time there was a decrease in the percentage of patients who felt the operation had made no difference, with a corresponding increase in the percentage of patients who felt worse. Although the number of patients who benefited remained fairly constant, there was deterioration in the quality of benefit in the first year, after which the benefit remained stable. This has implications in the pre-operative counselling of patients. PMID- 11027051 TI - Evaluation of functional outcomes (speech, swallowing and voice) in patients attending speech pathology after head and neck cancer treatment(s): development of a multi-centre database. AB - Since April 1997, in Melbourne, Australia, speech pathologists have collaborated to establish a prospective database of functional outcomes of speech, swallowing and voice for patients undergoing head and neck cancer treatments. Staff at eight acute care hospitals, all of which offer speech pathology for head and neck cancer services in Victoria, are contributing data, collated centrally, in an agreed pro forma. Early results are given (after 12 months' data collection). The implications for clinically-based research, and the future potential for benchmarking outcomes--by expansion of the rehabilitation database beyond the current participating sites--is discussed. This paper outlines the rationale of establishing the database is multicentered, and explores some of the complexities involved, including the challenges inherent in long-term accurate data collection in the head and neck cancer patient population. This work represents the development of an appropriate, usable tool for data collection on functional outcomes. PMID- 11027052 TI - Simple device for chemical cauterization of posterior bleeding points in the nose. AB - Epistaxis in adults often arises from a posteriorly-placed bleeding point. This paper describes a method of cauterizing these vessels under endoscopic control using simple materials found in most ENT departments, thus avoiding the need for more specialist equipment. PMID- 11027053 TI - Cerebellopontine angle lymphoma presenting as chronic mastoiditis. AB - Middle-ear tumours are not uncommonly mistaken for mastoiditis on presentation. We report a case of cerebellopontine angle non-Hodgkin's B-cell lymphoma originally presenting as a middle-ear inflammatory process. In mastoiditis not resolving with conventional treatment it is important to look for an underlying cause. PMID- 11027054 TI - Management of acute surgical orbital haemorrhage: an otorhinolaryngological and ophthalmological perspective. AB - This retrospective report describes the management and outcome of seven patients who suffered an acute surgical orbital haemorrhage (ASOH), secondary to a surgical procedure performed on either the sinuses, orbits or eyelids. All patients but one recovered their pre-operative vision. A management plan is outlined on how to assess and tackle this complication, so that the ENT surgeon may be better prepared to meet such an acute surgical emergency, should it arise. PMID- 11027055 TI - Primary chordoma of the ethmoid sinus. AB - Primary chordoma of the paranasal sinuses are extremely rare tumours, with only a small number of cases verified and published in the literature. It appears that only five of these cases involved the ethmoid sinuses either as a primary or by local invasion, and of these documented cases only one other has been found to solely involve the ethmoid sinus. We present a case of primary ethmoid sinus chordoma treated by wide local surgical excision and present a review of the literature with regard to prevalence and treatment rationale. PMID- 11027056 TI - Rare case of naso-oral fistula with extensive osteocartilaginous necrosis secondary to cocaine abuse: review of otorhinolaryngological presentations in cocaine addicts. AB - We report what we believe to be only the 10th case of palatal necrosis secondary to cocaine abuse in a 33-year-old female patient. Extensive necrosis also involved the cartilaginous and bony septum and paranasal sinuses. Following exclusion of other mid-line destructive diseases her treatment involved saline douches and cessation of cocaine. She remains under review within the department with no evidence of progressive disease. We present a review of the other nine cases of palatal necrosis reported in the world literature and demonstrate a greater incidence in female users. The various presenting conditions of cocaine abuse encountered within the head and neck region by the otorhinolaryngologist are then discussed. PMID- 11027057 TI - Pleomorphic adenoma causing acute airway obstruction. AB - A case is reported of a pleomorphic adenoma of the minor salivary glands of the oral cavity presenting with acute airway obstruction. This is the first reported case to our knowledge of a mixed salivary tumour of the upper respiratory tract causing upper airway obstruction and acute respiratory failure. The patient had to be intubated and transferred to the intensive care unit. After an elective tracheostomy was performed, the adenoma was excised from its fibrous capsule. It was found to originate from the soft palate and occupied the parapharyngeal space. A high index of suspicion should be kept in order to diagnose tumours of the parapharyngeal space with unusual presentation. These tumours which are usually benign should be considered in the differential diagnosis from more common infectious or traumatic conditions and surgical morbidity should be minimal. PMID- 11027058 TI - Post-cricoid web associated with Rubinstein-Taybi syndrome. AB - A case of dysphagia due to a post-cricoid web in a seven-year-old boy with Rubinstein-Taybi syndrome is presented. The main features of this syndrome are summarized with particular reference to dysphagia and with previous cases of post cricoid webs in children reviewed. PMID- 11027059 TI - Pleomorphic adenoma of the retropharyngeal space: a rare location. AB - A case of an isolated pleomorphic adenoma of the retropharyngeal space is reported, this has not been documented previously in the literature. Attention is drawn to the wide spectrum of benign and malignant neoplasms that can potentially occur within this complex anatomical region. The importance of a systematic and logical approach to the management of such lesions is emphasized. PMID- 11027060 TI - Pleomorphic adenoma involving the stylomastoid foramen. AB - A rare case of an intratemporal pleomorphic adenoma is presented and the management of such a tumour is discussed. Some anatomical aspects of the facial nerve, pertinent to the pathophysiology of facial paralysis are outlined. This case demonstrates that tumour extension into the temporal bone can be resected successfully at initial surgery with excellent facial nerve functional outcome. We advocate exploration of the fallopian canal to be carried out at primary surgery and be performed by a surgeon familiar with the surgical anatomy of the intratemporal segment of the facial nerve. This approach will reduce the risk of facial nerve injury and palsy both at surgery and subsequently. PMID- 11027061 TI - Plexiform neurofibroma of the submandibular gland. AB - A rare plexiform neurofibroma of the submandibular gland in a patient with neurofibromatosis is presented. The clinical manifestations of the disease are reviewed. The need for early diagnosis of neurofibroma is emphasized because of malignant transformation. PMID- 11027062 TI - Plasma cell polyp of the vocal fold. AB - Plasma cell polyps of the vocal fold (plasma cell granulomas) are rare inflammatory polyps of the larynx. They should be included in the clinical and histological differential diagnosis of laryngeal polyps. Histologically they are polyclonal aggregates of plasma cells. It is essential to distinguish them from monoclonal, neoplastic plasma cell proliferations. The treatment of choice is surgical resection, although radiotherapy, laser ablation, antibiotics and steroids have been used successfully. We present a case of plasma cell granuloma presenting as a vocal fold polyp, treated surgically. PMID- 11027063 TI - Dual infection with atypical mycobacteria and Mycobacterium tuberculosis causing cervical lymphadenopathy in a child. AB - The most common presentation of mycobacterial infection encountered in otolaryngological practice is cervical lymphadenitis. We report a child with an unusual cause of cervical lymphadenopathy, i.e. dual tuberculous infections. This had clinical ramifications as, initially Mycobacterium avium-intracellulare was grown in culture and was resistant to standard anti-tuberculous agents, and hence treated with excision of the lymph node. However, the cultures from the excised lymph node grew out Mycobacterium tuberculosis that was sensitive to standard anti-tuberculous drugs. To our knowledge, no such presentation has been reported previously. We also review the literature on cervical lymphadenitis due to atypical mycobacteria and Mycobacterium tuberculosis, with particular emphasis on clinical presentation, diagnosis and management. PMID- 11027064 TI - Dangerous liaisons: a social history of venereal disease in twentieth-century Scotland. PMID- 11027065 TI - From lesion to metaphor: chronic pain in British, French and German medical writings, 1800-1914. AB - This is the first monograph devoted to the history of chronic pain. A novel methodology is used. Examining responses to a problem that remained stable over time anchors a survey of shifting terms and theories and leaves the historical invariance of the clinical syndrome open to textual research. Writings by medical authors from a wide range of professional backgrounds are examined including surgeons, physicians, psychiatrists, neurophysiologists, neurologists and psychoanalysts. Early responses to the problem of chronic pain without structural lesion were the appearance of neuralgia, a neuro-anatomical rewriting of traditional sympathies, extension of the concept of lesion to embrace disturbance of function and appeals to cenesthesis. Later in the century distinctions were drawn between hysterical and neuralgic pain, and between ideogenic, psychogenic and neurogenic pain. Some argued for the physiological equivalence of chronic pain and melancholia, while pain was central to Freud's original notion of conversion. This evidence of continuous discussion of lesionless pain throughout the century challenges the orthodox historical view that the rise of neuroscience meant such pain was simply dismissed as imaginary. The historical invariance of a syndrome of chronic pain without lesion speaks against histories of lesionless syndromes premised on social constructionism. The historical findings are relevant to contemporary debates about the nosology and nature of chronic pain. PMID- 11027066 TI - Introduction: Malthusian selections. PMID- 11027067 TI - Malthus's political economy of health: the critique of Scandinavia in the Essay on population. PMID- 11027068 TI - Boys to men: moral restraint at Haileybury College. PMID- 11027069 TI - The Malthusian moment. PMID- 11027070 TI - 'Malthus on man--in animals no moral restraint'. PMID- 11027071 TI - Malthus among the theologians. PMID- 11027072 TI - Malthus and the doctors: political economy, medicine, and the state in England, Ireland, and Scotland, 1800-1840. PMID- 11027073 TI - Malthusian mutations: the changing politics and moral meanings of birth control in Britain. PMID- 11027074 TI - Reproduction and revolution: Paul Robin and neo-Malthusianism in France. PMID- 11027075 TI - Biology and sociology of fertility. Reactions to the Malthusian threat, 1798 1933. PMID- 11027076 TI - Imaging of acute abdominal emergencies in infants and children. PMID- 11027077 TI - Recent advances in the understanding of reproductive malignancies. A new section. PMID- 11027078 TI - Recent advances in gestational trophoblastic disease. AB - Recent advances have increased our understanding of gestational trophoblastic disease, and epidemiologic studies have demonstrated that there are important differences in risk factors for complete and partial mole. Complete moles are now increasingly being diagnosed in the first trimester, affecting their clinical presentation and pathologic characteristics. While important advances have been made in chemotherapy, it is now recognized that etoposide is associated with a risk of second tumors. Several studies have advanced understanding of the molecular biology of gestational trophoblastic disease, and this is important for the eventual development of new and innovative therapy. PMID- 11027079 TI - New technologies in gynecologic cytology. AB - New technologies are trying to make significant changes in the way we perform the Pap smear test, a test that has undergone no significant modification since its inception. Efforts have focused on improving sampling with liquid-based methods, improving detection of abnormal cells with automated screening and improving the detection of human papillomavirus with assays practical for use in routine clinical material. This article reviews the performance of the new technologies to date, with an emphasis on those that are currently approved for cervical cytology testing. In the aggregate, these technologies can have a positive impact on cervical disease detection and intervention. PMID- 11027080 TI - Oral contraceptives and epithelial ovarian cancer. Does dose matter? AB - OBJECTIVE: To determine the risk of ovarian cancer among women who use low estrogen-dose oral contraceptives. STUDY DESIGN: The study used data on white women under 70 years of age who had been enrolled in a population-based case control study conducted between 1986 and 1988 in three western Washington counties. Women with ovarian cancer (n = 276) were ascertained through a population-based cancer registry, and controls (n = 391) were selected by random digit dialing. Unconditional logistic regression was used to estimate the risk of ovarian cancer associated with oral contraceptive use. RESULTS: After adjustment for age and parity, women who took oral contraceptives for at least three months were at decreased risk of ovarian cancer (odds ratio [OR] 0.8, 95% confidence interval [CI] 0.5-1.1) relative to women who never used this form of contraception. The reduced risk of ovarian cancer was present among women whose only preparation contained a low (< 50 micrograms ethinyl estradiol or < 80 micrograms mestranol) (OR 0.6, 95% CI 0.3-1.1) and high (OR 0.8, 95% CI 0.5-1.2) estrogen dose. CONCLUSION: While our results are limited in their statistical precision and by the inability of many subjects to recall the brands of oral contraceptives that they took, they suggest that the newer, low-estrogen-dose oral contraceptives confer a benefit regarding ovarian cancer risk similar to that conferred by earlier, high-estrogen-dose formulations. PMID- 11027081 TI - Maternal smoking and preeclampsia. AB - OBJECTIVE: To study the relationship between maternal smoking and preeclampsia and whether this association differs between primiparous and multiparous women. STUDY DESIGN: We conducted a population-based, retrospective, cohort study of 58,216 singleton pregnancies from northern and central Alberta, Canada, between 1995 and 1997. Multivariate logistic regression was used to control for maternal alcohol consumption, drug dependence, maternal age, maternal weight, prior intrauterine growth restriction and other confounders. RESULTS: Maternal smoking was associated with a significantly reduced overall risk of preeclampsia (adjusted odds ratio [aOR]: .61; 95% confidence interval [CI]: .50-.75; P < .01). Stratified analyses showed that in primiparous pregnancies, maternal smoking was associated with a significantly decreased risk (aOR: .63; 95% CI: .50-.80; P < .01); in multiparous women, maternal smoking was not associated with a statistically significant decreased risk of preeclampsia (aOR: 0.72; 95% CI: .51 1.02; P > .05). CONCLUSION: Maternal smoking is protective against preeclampsia. Understanding the underlying biologic mechanisms of this protective effect may advance our knowledge of the pathogenesis of preeclampsia. PMID- 11027082 TI - Indications for and outcomes of emergency peripartum hysterectomy. A five-year review. AB - OBJECTIVE: To review risk factors, management and outcomes of emergency peripartum hysterectomy performed in the last five years at Stony Brook University Hospital. STUDY DESIGN: Retrospective descriptive and cohort analyses from January 1990 to January 1995. Incidences of emergency peripartum hysterectomy and placenta accreta were determined. Relative risks of hysterectomy for specified risk factors were calculated. RESULTS: There were 39 cases of emergency peripartum hysterectomy, for an overall incidence of 2.7/1,000 births. Indications for emergency hysterectomy were placenta accreta, unspecified bleeding, uterine rupture, myomas and atony with placenta accreta, the most common. The crude relative risk of emergency hysterectomy was 46.9 (n = 37) for cesarean delivery, 15.24 (n = 31) for prior cesarean delivery and 110.83 (n = 21) for placenta previa. CONCLUSION: Cesarean delivery, prior cesarean delivery, placenta accreta and uterine atony were identified as risk factors for emergency peripartum hysterectomy, and abnormal placentation was the primary cause of cesarean hysterectomy. PMID- 11027083 TI - Laparoscopically assisted vaginal hysterectomy. A suitable substitute for abdominal hysterectomy? AB - OBJECTIVE: To compare short-term clinical results in a retrospective case-control review of laparoscopically assisted vaginal hysterectomy (LAVH) versus total abdominal hysterectomy (TAH). STUDY DESIGN: One hundred seventeen women undergoing laparoscopically assisted vaginal hysterectomy were compared to 117 women undergoing total abdominal hysterectomy, with or without bilateral salpingo oophorectomy. The perioperative and postoperative courses of the matched groups were compared. RESULTS: The time intraoperatively was longer in the LAVH group. The mean blood loss was increased in the LAVH group. The need for blood transfusion was higher in the LAVH group. The number of women with intraoperative complications differed significantly between LAVH and TAH. The number of postoperative complications did not. Postoperative time in the hospital was shorter in the LAVH group. The cost of the average case, including operating room time, instruments and hospital stay, did not differ. CONCLUSION: LAVH offers benefits to patients in the form of less time in the hospital and presumably, therefore, faster recovery, though at the expense of potentially longer intraoperative time, increased risk of blood transfusion and increased risk of intraoperative complications. PMID- 11027084 TI - Pregnancy outcome and complications in women with spina bifida. AB - OBJECTIVE: To describe the antenatal complications, mode of delivery and outcome of pregnancy in women with spina bifida. STUDY DESIGN: Case series of women known to have attended the spina bifida clinic at the Royal Children's Hospital. Medical records, postal questionnaire and telephone interview were utilized to collect data on the effect of pregnancy on the health of women and the effect of spina bifida on pregnancy outcome. RESULTS: Of 207 women born between 1945 and 1975, 23 reported having a pregnancy, and 17 who had completed pregnancies agreed to participate. The 17 women had a total of 29 pregnancies, with 23 pregnancies progressing to births. Fourteen of 17 women had antenatal admissions, with wheelchair-dependent women requiring more-frequent and longer admissions. Recurrent urinary infections in pregnancy occurred in women with a prior history of urinary infections; stomal problems occurred but were not serious; mobility was reduced for two women during pregnancy, with full recovery afterwards; and preexisting pressure sores worsened during pregnancy. Vaginal deliveries occurred in one in five pregnancies of women who were wheelchair dependent and in ten of eighteen pregnancies in independently mobile women, including seven of eight pregnancies of independently mobile women without ileal conduits. Cesarean sections were accompanied by postoperative complications in 10 women. CONCLUSION: Women with spina bifida who become pregnant generally have a positive outcome, with relatively low complication rates. PMID- 11027085 TI - Pyogenic granuloma (pregnancy tumor) of the nasal cavity. A case report. AB - BACKGROUND: Pyogenic granuloma is a benign lesion of unknown etiology that occurs in the skin and mucous membranes. It has been reported as a common lesion of the oral cavity and, less commonly, the nasal cavity. Although its etiology is unclear, pyogenic granuloma has long been associated with pregnancy, oral contraceptives and trauma. The term pregnancy tumor has been used to reflect the association with pregnancy; nevertheless, pyogenic granuloma is often misdiagnosed in obstetric patients. CASE: We present the case of a 37-year-old, pregnant woman with an enlarging nasal mass ultimately diagnosed as pyogenic granuloma. The patient was treated successfully with endoscopic excision of the lesion. CONCLUSION: Pyogenic granuloma is a rapidly growing lesion that should be considered in the differential diagnosis of any gravid patient with a mass in the oral or nasal cavity. The lesion may result in complications of the pregnancy, and management should be individualized for each patient. PMID- 11027086 TI - Methotrexate for cesarean scar pregnancy after in vitro fertilization and embryo transfer. A case report. AB - BACKGROUND: Cesarean scar pregnancy is an exceedingly rare occurrence. We present the first case of cesarean scar pregnancy following in vitro fertilization-embryo transfer (IVF-ET). CASE: A 40-year-old woman with a history of a previous cesarean section presented with five years of unexplained infertility. The patient complained of abdominal pain 16 days after embryo transfer. Ultrasonography revealed a gestational sac with cardiac activity located outside the lower segment of the uterus. Dilatation and curettage was performed due to misdiagnosis of inevitable abortion. Two weeks later, repeated sonography demonstrated a sacculus, 4.07 x 4.07 cm, within the uterine isthmus with only 7.1 mm of thickness separating the sac from the urinary bladder. Normal cervical length without ballooning was noted. Cesarean scar pregnancy was diagnosed. Local injection of methotrexate (MTX) under ultrasound guidance was performed. Plasma beta-hCG levels declined from 23,328 to 8 mlU/mL within two months. CONCLUSION: For women with cesarean scar pregnancy who desire fertility, conservative treatment using MTX is an excellent choice. PMID- 11027087 TI - Adenocarcinoid tumor of the ovary diagnosed during pregnancy. A case report. AB - BACKGROUND: Adenocarcinoid tumors are uncommon neoplasms with dual morphology, showing components of a neuroendocrine tumor with carcinoid features and an adenocarcinomatous component composed of glands lined with mucin-containing cells, some of which are goblet type. CASE: A 36-year-old woman had a left adnexal mass found during the second week of pregnancy. Sonography showed it to be increasing in size and eventually to become associated with pelvic pain. During the 20th week of gestation, an exploratory laparotomy was performed, and the left ovary and fallopian tube were excised. A diagnosis of adenocarcinoma was rendered by intraoperative frozen section. A staging procedure was then performed that included removal of the contralateral adenexa, pelvic lymph node sampling, peritoneal biopsies and partial omentectomy. The vermiform appendix and gastrointestinal tract appeared unremarkable. The patient was discharged. Permanent sections of the left ovary revealed an adenocarcinoid tumor. CONCLUSION: While reports detail ovarian metastases of adenocarcinoid neoplasms from primary appendiceal and other gastrointestinal sites, this case, in the setting of a normal appendix and negative workup for an extraovarian origin, is the fourth documented one of a primary ovarian adenocarcinoid tumor and first diagnosed during pregnancy. PMID- 11027088 TI - [Drawing of the family and of the human figure: a comparative study between children with epilepsy and a control group]. AB - OBJECTIVE: In this study our aim was to get to know children with epilepsy through their drawings. In this case the drawings of the human figure and the family were chosen. PATIENTS AND METHODS: The population which was studied included 32 children: 16 of these children had epilepsy and the 16 others were the control group. RESULTS: From the comparison performed between these two groups of children it was found that in relation to family drawings children with epilepsy tend more to draw their own family than children in the control group. As for the drawing of the human figure and the global analysis of the body figure it was found that the mental age of the isolated character and the higher level character drawn by epileptic children was inferior to chronological age, while the reverse was true in the drawings of the control group. In relation to the analysis of structural cohesion in the representation of the body it was found that hands were absent in the drawings of epileptic children. PMID- 11027089 TI - [Cerebral hemodynamics in the syndrome of pseudomigraine with csf-pleocytosis:a transcranial doppler study]. AB - INTRODUCTION: The clinical picture of the pseudomigraine with CSF pleocytosis syndrome is of intermittent headaches, sometimes accompanied by episodes of transient neurological defects and lymphocyte pleocytosis, which occur for a variable length of time, followed by spontaneous resolution. It has been suggested that the cause is immunogenic due to post-infectious activation of the humoral immune system against the leptomeningeal vascular structures, but this hypothesis does not explain the accompanying transient deficit phenomena. MATERIAL AND METHODS: We present a study of cerebral hemodynamics using transcranial Doppler in a patient diagnosed as having pseudomigraine with pleocytosis according to current criteria. RESULTS: Our results suggest the presence of cerebral vascular instability and increased vascular reactivity to induced hypocapnia and hypercapnia, similar to that reported in patients with migraine with aura. At no time was significant increase in the average rate of cerebral blood flow detected, comparable to that usually seen in patients with bacterial or viral meningitis. CONCLUSIONS: Although the cause cannot be established, from our study it would seem that headache in the pseudomigraine with pleocytosis syndrome is related to cerebral vascular instability and therefore probably to activation of the trigeminal-vascular system. It is possible that the deficit phenomena may also have a similar explanation to that of patients with migraine with aura, that is, a propagated cortical depression. Transcranial Doppler studies may be useful in the diagnosis of pseudomigraine with pleocytosis syndrome. PMID- 11027090 TI - [Indications for carotid endarterectomy without arteriography. A validation study using eco-Doppler]. AB - INTRODUCTION: The possibility of diagnosing carotid stenosis and carrying out surgery without arteriography has increased with the use of different diagnostic methods. The eco-Doppler has been shown to be a useful method, although it requires previous individualised validation. OBJECTIVE: We aim to validate our Vascular Diagnostic Laboratory in the diagnosis of stenosis of the carotid bifurcation using eco-Doppler as compared with angiography, and the therapeutic indication thus obtained. PATIENTS AND METHODS: We made a prospective study for one year of 62 consecutive patients diagnosed on eco-Doppler as having carotid stenosis of over 70% and subsequent carotid arteriography. The treatment indicated is given, without waiting for the result of the angiography, on clinical evaluation, computerized tomography or cranial magnetic resonance and eco-Doppler, and subsequently on angiography. The degree of correlation of both methods, both for the therapeutic indication and for the degree of stenosis was determined. RESULTS: In five cases there was discrepancy and the therapeutic indication was different. Three of these were related to the diagnosis of carotid occlusion and in the other two cases there were bilateral lesions of over 50% on eco-Doppler, which were classified as minor on angiographic study. In the group with stenosis of over 70% with < 50% contralateral stenosis, eco-Doppler showed sensitivity and specificity of 100%, with a kappa correlation index = 1. CONCLUSION: It is possible to indicate carotid endarterectomy in patients with unilateral stenosis greater than 70% and contralateral stenosis < 50%, based on the eco-Doppler studies done in our Vascular Diagnostic Laboratory. PMID- 11027091 TI - [Eosinophilic meningoencephalitis in Villa Clara (Cuba). A study of 17 patients]. AB - INTRODUCTION: Infection by Angiostrongylus cantonensis is caused by eating snails and slugs or contaminated vegetables, and is the commonest cause of eosinophilic meningoencephalitis throughout the world. OBJECTIVES: To make a further study of this condition, analyze the clinical behaviour and evaluate the cerebrospinal fluid findings and response to treatment. PATIENTS AND METHODS: We made a retrospective, descriptive study by reviewing the clinical histories of a series of 17 adult patients attended in two university hospitals in the province of Villa Clara (Cuba), between December 1991 and January 2000. We recorded the characteristics of the clinical picture, results of the complementary investigations, treatment given and clinical course and the necropsy findings of the three patients who died. The data was collected from questionnaires. RESULTS: Headache was a constant symptom. Twelve patients had high temperatures and seven had stiff necks. Motor deficit and coma were seen in the three patients who died. In two patients the cranial nerves were involved. Spinal fluid pleocytosis varied between 48 and 2570 cells/mm3. Initially there was lymphocyte predominance in 10 patients and subsequently eosinophil conversion. The peripheral eosinophilia, seen in all patients, varied between 7% and 61%. Of the 14 patients cured, eight received symptomatic treatment and six antihelminth drugs. Steroids were given to only one patient. Of the three patients who died, two had received antihelminth drugs. CONCLUSIONS: Motor deficit and coma worsen the prognosis, which is generally good. There is no relation between cerebrospinal fluid pleocytosis and raised protein levels in the cerebrospinal fluid, nor between blood eosinophilia and prognosis. The most difficult differential diagnosis in our patients was with leptospirosis. No differences were seen in clinical evolution in relation to the treatment given. PMID- 11027092 TI - [Invasive diagnosis versus non-invasive diagnosis in carotid disease: an argument out of focus]. PMID- 11027093 TI - [Melanocytic schwannoma. A case report and review of the literature]. AB - INTRODUCTION AND CLINICAL CASE: We report a new unusual case of a patient without known familiar history of melanocytic schwannoma or Carney complex with multiple melanocytic schwannoma in the spinal canal. Histologically, the tumor was composed of irregular interlacing spindle-shaped cells with pigments. The diagnosis is made by immunohistochemical techniques and electron microscopy findings. CONCLUSION: We review the literature, and the different theories on the origin of these tumors are discussed. PMID- 11027094 TI - [Acute transverse myelitis secondary to hepatitis B vaccination]. AB - INTRODUCTION: Acute transverse myelitis is an inflammatory disorder. The pathogenesis is unclear, but the probable mechanism involves an autoimmune phenomenon. Possible causes included multiple sclerosis and parainfectious and postvaccinal events. Myelitis has rarely been reported secondary to vaccinations including hepatitis B. We present a case of acute myelitis, which seems secondary to the administration of the hepatitis B vaccine. CLINICAL CASE: A 15-years-old female presented with progressive numbness of the right arm and leg, with right leg weakness. Symptom began one week after receiving the first dose of the hepatitis B vaccine. Spinal cord magnetic resonance (MR) revealed a diffuse increased signal extending from C6 to D2. Cerebral MR and cerebrospinal fluid were normal. She was treated with high doses of methylprednisolone with a complete recovery of neurological functional. Repeat medullar cord MR was normal. There was no relapse during a four years follow up. CONCLUSIONS: Potential causal relationship between vaccination against hepatitis B and multiple sclerosis was brought to the attention and to public debate. However, no conclusive association could be made between vaccination and demyelination. In the clinical setting, the distinction between a first episode of multiple sclerosis or postvaccinal myelitis depends upon subsequent course. PMID- 11027095 TI - [Sporadic familial meningiomas]. AB - INTRODUCTION: Meningiomas are primary tumours of the central nervous system. Usually they are sporadic. The occurrence in more than one member of a family is unusual. Up till now this coincidence had been related with type 2 neurofibromatosis. In this paper we comment on two siblings who did not fulfil neurofibromatosis diagnostic criteria as an example of sporadic familial meningiomas. CLINICAL CASES: Two siblings (a 79 years old female and a 77 years old male) were diagnosed of a meningioma in an interval of two years, with surprising clinical and neuroimaging similarities. In sporadic meningiomas, abnormalities in the long arm of chromosome 22 have been found. Type 2 neurofibromatosis causative gene has also been found in this chromosome. Meningiomas are quite often found in this entity, and therefore, this gene was implicated as a main factor in the genesis of an important number of meningiomas. However, several studies have not found an association between these tumours and the locus for neurofibromatosis, leading to think that there may be other genes that may influence on meningiomas development. PMID- 11027096 TI - [Sudden unexpected and unexplained death in epilepsy]. AB - INTRODUCTION: Sudden unexpected death in epilepsy (SUDEP) is a rare and disconcerting event, scarcely reported in our country. DEVELOPMENT: The definition proposed at the last meeting of experts, in 1997, excludes all cases with a known cause of death, except for the seizure itself or the antecedent of epilepsy. The cause-effect relationship between epileptic seizure and sudden death has not been definitely proved but relies heavily on the following facts: 1. Epidemiologic studies: the incidence of SUDEP is larger in severe epilepsy, as shown by hospital records, than in epileptics of the general population; increases proportionately with the increasing requirements of antiepileptic polytherapy, and with raised seizure frequency (except for Idiopathic Epilepsy, where this fact has not been documented). 2. Physiologic studies in humans during seizures, suggest that in some cases a central apnea occurs, occasionally followed by asystole; in some others, cardiac arrhythmia, of reflex neural origin, have been detected. Animal experiments support the first hypothesis. 3. The latest pathologic studies at autopsy support also the second hypothesis: cardiac ischemia due to repeated vasospasms, neurally activated, during seizures. A causal relationship of SUDEP with antiepileptic drugs administration has not been proved, but the sudden decrease of antiepileptic drugs serum levels may cause cardiac arrhythmias potentially fatal. Differential diagnosis with certain cardiopathies should be performed. CONCLUSION: For the time present, only a better knowledge of the risk factors of SUDEP will allow us to reduce its incidence. PMID- 11027097 TI - [Clinical value of EEG coherence as electrophysiological index of cortico cortical connections during sleep]. AB - OBJECTIVE: The technique of EEG coherence gives a non-invasive objective index of the functional relations between the different regions of the cerebral cortex. The main objectives of this study were firstly to obtain methodological information to facilitate its use in any context, whether clinical or in investigation, and secondly to draw attention to the results obtained with this technique during the different phases of sleep, both in healthy persons and in certain clinical conditions where its use in evaluation and/or diagnosis has been shown. DEVELOPMENT: The results derived from the application of coherence analysis during normal sleep show the existence of a high level of interhemisphere connectivity during all phases of sleep. This result has fomented its use in the evaluation and diagnosis of disorders in which it is suspected that there are anomalies in the relation between the two hemispheres, such as neurological changes (agenesis of the corpus callosum, AIDS), psychiatric disorders (depression) and in the prognosis of states of coma. Also, progressive neurone degeneration and neurochemical deficits characteristic of Alzheimer-type dementia suggest that the coherence EEG during the REM phase may give electrophysiological indices in the establishment of the diagnosis and prognosis of the disorder. CONCLUSION: This study shows that the coherence technique is a useful tool when establishing the pattern of cortico-cortical interactions subjacent to different functional states of the brain, both in healthy persons and in those with the different diseases in which there are alterations in cerebral function. PMID- 11027098 TI - [Spatial learning and the hippocampus]. AB - INTRODUCTION: Since publication of the book The hippocampus as a cognitive map by O'Keefe and Nadel in 1978, several theoretical and experimental works have supported the participation of the hippocampus in rodents in spatial processing. Since then, great advances have been made into understanding the potential role of the hippocampus in learning and memory processes, relegating a secondary role to the hypothesis that, in rodents, the hippocampus is restricted to the formation of a cognitive map. DEVELOPMENT: In this work, different forms of spatial navigation are analyzed (taxis, cartographic and route integration-based navigation) on a psychological and neurobiological level. The spatial function of the hippocampus proposed by O'Keefe and Nadel is discussed but a more general model is chosen that explains the function of the hippocampus in mammals, i.e. a model of relational memory. Therefore, in the context of relational memory we attempt to explain the different forms of spatial memory in rodents and present the most relevant experimental data that support a critical role for the hippocampus in this kind of memory. CONCLUSIONS: The studies reviewed in this work support an important role for the hippocampus in spatial processing. However, this does not appear to be its sole function and it possibly operates more widely also participating in relational memory. PMID- 11027099 TI - [Structural review of chemical agents that can induce parkinsonism]. AB - OBJECTIVE: To identify similar structural elements in the largest number possible of chemical agents which can induce Parkinsonism and study their relation with the main chemical changes seen in Parkinsonism. DEVELOPMENT: We studied the chemical structure and structural relations of the chemical agents reported in the literature because of their ability to induce Parkinsonism. We observed the structural similarity of the majority of chemical agents and the structural discrepancy of the rest. The structural element identified as common had three clearly differentiated parts: a cyclic fragment, bridge fragment and nitrogen atom fragment. We studied the diversity of the parts of each fragment. We studied the relation of the common structural element with other biochemical components which are altered in Parkinsonism, and also the possible mechanism of action of the structural element in the process of induction of Parkinsonism. CONCLUSIONS: There is a structural grouping close to the molecule of the dopamine neurotransmitter in most of the organic compounds which can induce Parkinsonism. Therefore, we suggest that the origin of certain Parkinsonian conditions may be due to changes in the natural equilibrium of dopamine due to the action of chemical agents with a suitable chemical substructure similar to that of dopamine. PMID- 11027100 TI - [The treatment of multiple sclerosis with beta-interferon 1a]. AB - INTRODUCTION AND OBJECTIVE: Preliminary studies (phase III) have shown that treatment with beta-interferon 1a (avonex) is effective in diverse aspects of relapsing/remitting multiple sclerosis (RRMS). The present study discusses results of an opened prolonged (phase IV) trial at the Baylor/Methodist International Multiple Sclerosis Center using 6 MIU by intramuscular injection once a week. PATIENTS AND METHODS: Clinical measurements: EDSS and Patient's Subjective Self-Evaluation (PSSE) and magnetic resonance (MR) were utilized every three months in 300 patients studied between 1996-1999. Patients with RRMS (71.8%) and secondarily progressive multiple sclerosis (SPMS) (28.2%) were included. RESULTS: Global clinical improvement parameter integration was 22.7% and clinical stabilization 39.4%. MR did not show quantitative alterations for either group. Sixteen patients abandoned treatment and 16 (5.3%) had relapses correlated by MR activity; 34.9% remained stable by EDSS and magnetic resonance findings but deteriorated subjectively by PSSE (62% has SPMS). CONCLUSION: This phase IV study indicates that prolonged treatment of RRMS and SPMS with avonex maintains a positive effect in 62.1% of patients. PMID- 11027101 TI - [Immunological mechanisms involved in the pathogenesis of multiple sclerosis]. AB - OBJECTIVE: To summarize current knowledge of the immunological mechanisms involved in the pathogenesis of multiple sclerosis and its implications in the development of new therapeutic options. DEVELOPMENT: The aetiology of multiple sclerosis is still not completely clear. Nevertheless there is much evidence in favor of the possibility that autoimmune phenomena play an important part in the development and course of the disorder. Knowledge of the immunological phenomena involved determines the possibility of modifying the immune response at different levels. Thus it is possible to act on: 1. The trimolecular complex during lymphocyte activation. 2. To block the passage of activated cells across the blood-brain barrier. 3. To modify the effector action of specific antigen cells, and 4. Induce populations of regulator cells. Similarly, immunological phenomena which develop during pregnancy and the postpartum period are an interesting physiological model which permit identification of the mechanisms by which autoimmune disorders are modified. Amongst the factors affecting the immune response during pregnancy and the postpartum period the various maternal and foeto-placental hormones should be considered. Their actions may be due to their effects on the secretion of different groups of cytokines. Better understanding of the factors regulating the immune response during pregnancy and the postpartum period permits definition of new potential sites for therapeutic action. CONCLUSIONS: Current knowledge of the many physiopathological mechanisms involved in the development of multiple sclerosis allows rational development of new alterative treatments for this condition. PMID- 11027102 TI - [Neurorehabilitation of multiple sclerosis]. AB - INTRODUCTION: Multiple sclerosis (MS) is the second most important/common cause of neurological disability in young adults. Current treatment includes the new immunomodulating agents and various drugs used in symptomatic management. Neurorehabilitation (NR) complements these treatments, forms an integral therapeutic strategy and is orientated to improve the activity and participation of the patient from a biopsychosocial point of view. DEVELOPMENT: Although there is a generally favorable opinion towards NR, there is little scientifically validated evidence regarding the benefits of its use. Correct measurement of the results is essential for their interpretation in terms of efficiency and efficacy. Of the various instruments for measurement of the results of NR, there are scales which have been shown to be of satisfactory clinical usefulness and scientifically sound for use in NR programmes in MS. In spite of methodological difficulties for the design of protocols in this field, in recent years several well-designed studies (randomised, controlled, double or single blind) have shown the usefulness of NT in MS (both outpatients and inpatients and including progressive forms) since handicap has been reduced and quality of life improved. These studies refer to NR as a multidisciplinary approach (a specially trained team of professionals) with concrete objectives and established job descriptions, a suitable battery of instruments for measurement of the results, active participation of the patient and his family and correct follow-up in the community. PMID- 11027104 TI - [A review of the epidemiology of multiple sclerosis in Mexico]. AB - OBJECTIVE: To review the epidemiological information on multiple sclerosis in Mexico. DEVELOPMENT AND CONCLUSIONS: The results of the few epidemiological studies on multiple sclerosis show a prevalence of 1.5/100,000 inhabitants, a figure which is not in concordance with studies carried out by institutions, which indicates an increase in the frequency of new cases. This information is not necessarily due to a true increase in prevalence. Possibly this increase is related to improved methodology for clinical and paraclinical diagnosis, especially the use of magnetic resonance. There is a need for descriptive and analytical epidemiological studies to obtain precise data on this. At present, the Mexican Group for the study of Multiple Sclerosis and The Mexican Association for the study of Multiple Sclerosis are using a questionnaire designed and validated for Mexico in order to determine the clinical characteristics and the prevalence of multiple sclerosis. The results will be available in the near future to improve knowledge regarding the epidemiology of this disorder and its economic impact on the Mexican health systems. PMID- 11027103 TI - [Actual status of multiple sclerosis in Cuba. National cooperative group for the clinical trial "Recombinant interferon alpha-2b in multiple sclerosis"]. AB - INTRODUCTION: Multiple sclerosis (MS) has been considered to be an uncommon disorder in Cuba. In recent years many Cuban investigators have worked hard studying MS in Cuba. DEVELOPMENT: We review the history, epidemiology, clinical characteristics, complementary investigations and clinical trials of recent studies published in national and international journals on the most important investigations done on MS in Cuba. There is an outstanding placebo-controlled, double blind, multicentric randomised clinical trial using recombinant alpha 2b IFN in the relapsing-remitting clinical form in which the results obtained in the first 30 patients who completed 2 years treatment show a statistically significant reduction in the frequency of relapses and the number of patients free of relapses in patients who received alpha-IFN as compared with those given placebo treatment. CONCLUSIONS: There has been an increase in the number of studies made on MS in Cuba in the past two years. Numerous clinical, neuroepidemiological, neurophysiological, biochemical, immunological, imaging and neuropsychological investigations and clinical trials indicate a marked increase in understanding of MS. The results of all these studies indicated the need for the creation of a National Reference Centre for Patients with Multiple Sclerosis, due to the complexity of this type of patients and the need to offer improved medical attention from a specialist team and to integrate the investigations in Cuba and internationally so as to reach the level of more developed countries. PMID- 11027105 TI - [Late onset familial myasthenia gravis]. PMID- 11027106 TI - [Sella metastases mimicking hypophyseal adenomas]. PMID- 11027107 TI - [The locked-in syndrome: an ethical challenge]. PMID- 11027108 TI - [Neonatal hypotonia in Prader-Willi syndrome due to t(15;16)]. PMID- 11027109 TI - Sexual arousal in rapists as measured by two stimulus sets. AB - Some uncertainty exists in the literature regarding the status of phallometric testing with rapists. Although Quinsey and colleagues (1981) argue that rapists can be distinguished from nonsexual offenders with appropriate phallometric tests, Marshall (in press) argues that the validity of such testing is not proved, and that findings supportive of the testing are a artifact of population differences (i.e., psychiatric vs. correctional) and not reflective of rapists overall. The current study attempts to clarify this issue by testing rapists, child molesters, and mixed offenders (those with both adult and child victims) in a prison setting with both the Barbaree and Quinsey stimulus sets. It was hypothesized that rapists would be found to have more deviant rape indices when tested with the Quinsey stimulus set than with the Barbaree set. However, neither stimulus set distinguished the three groups in terms of the rape indices, whereas the rape index calculated from the Quinsey stimulus set was slightly more deviant than the one calculated from the Barbaree set. However, only 25% of rapists were classified as deviant using a rape index cutoff of 1.0. This finding is discussed in terms of the sexual preference hypothesis. PMID- 11027110 TI - The nature of sexual offenders' affective empathy: a grounded theory analysis. AB - The aim of this research was to investigate the nature of sexual offenders' affective empathy. Thirty-one men participating in a residential treatment program for child abusers constructed "victim apology letters" as a way of measuring/examining empathy deficits. The task was videotaped, transcribed, and subject to grounded theory techniques. It was discovered that intrafamilial offenders were more likely to minimize their behavior while exhibiting illicit power and control, whereas extrafamilial offenders were more likely to directly blame their victims and exhibit overtly explicit offense detail. From these open coded categories, the axial categories of self as nonoffender, external blaming, and secondary victimization were derived. These results may have implications for the delivery of victim empathy components of sexual offender treatment programs. PMID- 11027111 TI - Alcohol and drug abuse in sexual and nonsexual violent offenders. AB - According to a literature review by Marshall (1996), presently available data have not clarified the proportion of sexual offenders who would meet diagnostic criteria for addictive problems. Our own literature review failed to find published studies comparing sexual offenders to nonsexual violent offenders on standardized measures of substance abuse. Our study is a preliminary investigation of the differences between sexual offenders (rapists: n = 72; child molesters: n = 34) and nonsexual violent offenders (n = 24) on measures of alcohol and drug abuse. The findings indicate that sexual offenders were more likely to abuse alcohol than were nonsexual violent offenders. Nonsexual offenders were significantly more likely to have had a history of other forms of substance abuse. The results are considered in terms of theories of alcohol's contribution to aggressive behavior and sexual aggression. Implications for assessment and treatment of sexual offenders are discussed. PMID- 11027112 TI - Predictors of sexual recidivism: did meta-analysis clarify the role and relevance of denial? AB - Seven studies selected for meta-analysis of the denial variable in a large-scale meta-analysis of predictors of sexual recidivism are briefly reviewed and a number of methodologic issues are identified that relate to interpretation of the failure of meta-analysis to find an effect for the denial variable. These include variability in definition of the denial variable, variation in treatment access and exclusion for deniers, low base rates of recidivism, small sample sizes, low power, and high probability of type II error across individual studies. Despite the apparent objectivity and power of this methodology, a careful examination of the individual studies suggests that meta-analysis did not clarify the role and relevance of denial as a predictor of sexual recidivism. Further work is necessary to clarify the role and relevance of denial in treatment success, risk assessment, and risk prediction. PMID- 11027113 TI - A cautionary note regarding Nicholaichuk et al. (2000). AB - The treatment outcome study by Nicholaichuk, Gordon, Gu, and Wong (2000) used a novel method for identifying a comparison group of untreated sex offenders (i.e., drawing from existing criminal history records). A potential problem with their approach is that older records would be expected to include a disproportionate numbers of recidivists. Such an artifact is identified in Nicholaichuk et al.'s (2000) study; nevertheless, their data continue to suggest a small, positive effect for treatment even after eliminating the cases in which bias is most likely. PMID- 11027114 TI - Counselling in general practice. AB - Up to 30% of the problems that patients consult their GPs about have a psychosocial component. Managing the needs of these patients has contributed to the increase of counselling provision in primary care. Most primary care groups (81%) have some provision for practice-based counselling, and around a third to a half of general practices now have counselling services provided by a range of practitioners. Here, we assess the effectiveness of counselling for managing emotional and mental health problems in general practice. PMID- 11027115 TI - Leflunomide for rheumatoid arthritis. AB - Disease-modifying antirheumatic drugs (DMARDs) are given to patients with rheumatoid arthritis (RA) to prevent synovitis, slow destruction of articular cartilage and bone, preserve function and control systemic manifestations of the disease. Recognition that irreversible joint damage often occurs early in RA has led to much prompter use of DMARDs, with sulfasalazine or methotrexate commonly considered the treatment of first choice. Leflunomide (Arava-Aventis) is a new DMARD, licensed for the treatment of adults with active RA. The manufacturer claims that leflunomide has "comparable efficacy to methotrexate and sulphasalazine", with a "faster onset of action", and an "acceptable tolerability profile". Here, we consider the place of leflunomide in the management of patients with RA. PMID- 11027116 TI - Capturing prescribers' minds. AB - The prescription plays a key role in the provision of healthcare, culminating many consultations, affecting patient well-being and committing expenditure of around 13% of the NHS budget. Ultimately, the contents of the prescription depend on the prescriber's knowledge, attitudes and beliefs, and there are many who would wish to influence these. The eighth annual Drug and Therapeutics Bulletin symposium, 'Capturing prescribers' minds', looked at ways of influencing prescriber's decisions. Here, we summarise the key points raised in the presentations and in subsequent discussion. PMID- 11027117 TI - Complex systems in technology and policy: telemedicine and telecare in Japan. AB - We have examined aspects of the development of telemedicine and telecare in Japan. Despite Japan's reputation as an integrated and technocratic country, the diffusion of telemedicine has fallen below expectations, notwithstanding the urgent need to solve the problems of an ageing society. There has been a fragmentation of objectives and perspectives within the corporate (manufacturing) sector and within the policy sector of central and local government. There has also been a broader fragmentation between manufacturers, government and users (doctors and hospitals). As a result, the success or failure of individual projects appears to depend not on the quality of the technology, and often not on the evident importance of the social need, but on the overall coherence of a complex socio-technical system. PMID- 11027118 TI - The feasibility of telemedicine for orthopaedic outpatient clinics--a randomized controlled trial. AB - We investigated the use of videoconferencing in the examination of orthopaedic outpatients. A consecutive sample of orthopaedic outpatients was randomized to examination either via videoconferencing (n = 76) while attending a primary-care unit or at a conventional hospital outpatient clinic 160 km away (n = 69). Videoconferencing was found to be feasible and the equipment functioned well technically. There were somewhat more problems in examining the telemedicine patients than the clinic patients. The two patient groups were equally satisfied with the specialist service. The telemedicine patients were more willing to have their next visit by videoconferencing than the conventional patients. Videoconferencing between primary and secondary care can be used in the examination of orthopaedic patients whenever no demanding imaging technology is needed. PMID- 11027119 TI - Do community alarm users want telecare? AB - Previous surveys have suggested that community alarm users are in general satisfied with the service provided. However, there is also evidence that users would like additional options that could be provided by a telecare system. Inner city users of community alarms were consulted about their current systems and the extension of these to incorporate telecare options. The users lived in 160 homes in three multistorey apartment blocks. The average age of the 176 users surveyed was 76 years. Seventy-seven per cent of users were interested in automatic fall detection, 68% in lifestyle monitoring, 57% in telemedicine and 46% in videoconferencing. Nineteen of those interviewed (11%) did not want any of the suggested enhancements. The survey showed that elderly people are not technophobes, since 58% possessed either a microwave oven or a video-recorder, and also showed that the majority were prepared to accept new technologies that would support their independence. PMID- 11027120 TI - Transtelephonic transmission of electrocardiograms in Slovenia. AB - The transtelephonic transmission of electrocardiograms (ECGs) was introduced in Slovenia in 1996 and has been used regularly since September 1997 by health institutions and patients with cardiovascular diseases. Portable 12-channel ECG equipment weighing less than 200 g is generally used. The diagnostic centre is located in the intensive-care unit at the University Medical Centre Ljubljana. In the first 12 months of regular operation we received 463 calls, 213 of which were diagnostic. The most common reasons for calling were: ischaemic chest pain, atypical chest pain, palpitations and dyspnoea. The most common diagnoses made on the basis of history and the ECG were: acute myocardial infarction, angina pectoris, paroxysmal tachycardias and atypical chest pain. In 40% of cases the cardiologist's advice sufficed; 38% of patients were referred to the emergency room, 29% immediately and 9% only if a suggested therapy did not prove successful; and in 21% of cases new medication or a change of dosage of current medication was suggested. Transtelephonic ECG transmission enables direct communication between general practitioner or patient and cardiologist. With its use, time from onset of symptoms to initiation of treatment is shortened, which reduces disability and mortality due to cardiovascular diseases and improves cost benefit. PMID- 11027122 TI - An evaluation of the telepsychiatry programme in Victoria, Australia. AB - We evaluated the use of telepsychiatry in Victoria, a state of Australia with some five million people. There were 39 sites at the time of the survey, 12 metropolitan and 27 rural. Interviews and postal questionnaires were used. A total of 99 staff were surveyed, 69 by post and 30 during visits. There were 37 participants from metropolitan sites and 62 from rural sites. Rural sites conducted an estimated 137 sessions per month (i.e. an average of 5.1 per month per site). The majority (70%) of the usage in Victoria was for clinical purposes, both primary and secondary consultations. PMID- 11027121 TI - An evaluation of satisfaction with telemedicine among health-care professionals. AB - A survey was conducted among non-doctor health-care professionals in six rural counties in Missouri. The purpose of the survey was to establish baseline data to evaluate the effect of changes in the health-care sector, especially technology changes, on the job satisfaction, career satisfaction, relationships and communication activities of health professionals. The survey included three rural counties in which integrated telecommunication and interactive video telemedicine services were being installed, but before significant activities had begun, and three comparator counties without substantial integrated telecommunications infrastructure and telemedicine services. During a one-month study period, 1108 questionnaires were distributed. The total response rate was 50.1% (n = 555). Of the respondents, 30.3% indicated that technology in health-care was having a large effect on their work, although only 18.2% indicated that telemedicine and telecommunications were having a large effect. No systematic differences were found among the health professionals in the two communities at the time telemedicine equipment was being installed. PMID- 11027123 TI - The length of time necessary to break even after converting to digital mammography. AB - The cost differences between film-based mammography (FBM) and digital mammography (DM) were estimated after discussions with hospital personnel and industry representatives. Human resource costs were not included. The fixed cost of FBM per machine was estimated to be $50,000 and the variable cost $4.60 per examination. The fixed cost of DM per machine was estimated to be $102,000 and the variable cost $0.10 per examination. The total number of examinations required to break even was therefore 11,556. At a rate of 15 examinations per machine per day and with 251 working days per year, it would take 3.1 years to break even. In the first year after the break-even point had been attained, $16,943 would be saved for every 3765 examinations performed. Extrapolating to the USA as a whole, in which 23 million mammographic examinations are performed each year, suggests that the annual savings from going filmless would be more than $103 million. PMID- 11027124 TI - A pilot study of telemedicine for new neurological outpatient referrals. AB - We investigated whether new patients attending a neurological outpatient clinic could be safely managed by neurologists at a distance, using a video-link. In Northern Ireland, a video-link, transmitting at 384 kbit/s, was set up between a neurological centre and a small rural hospital 140 km away. Twenty-five unselected patients who had been referred by their family doctor were assessed by a neurologist using the telemedicine link and then immediately by another neurologist face to face. Examiners were blinded to the results of each other's assessment. In 24 cases the diagnoses made after the telemedicine and face-to face examinations were identical. There were minor differences between the type and number of investigations requested, and the requirements for treatment and follow-up between the two groups. Disposal method was the same in 21 of the cases. No major organizational difficulties were encountered during the study. The study showed that neurologists can deliver outpatient neurological care to distant patients using telemedicine. This has the potential to allow access to assessment for the large number of neurological outpatients who might otherwise be denied it. PMID- 11027125 TI - Video-capture teleradiology for the after-hours reading of computerized tomography scans. AB - We evaluated the after-hours support that radiologists could provide from their homes to an urban hospital for reading emergency computerized tomography (CT) scans. During a six-month study period, 36 CT scans were acquired using a video capture card in a PC and transmitted via an ordinary telephone line. The teleradiology interpretation and the formal report (from the hard copy) were compared with a reference or gold standard interpretation made by a radiologist and an emergency physician for 31 of the 36 cases. In comparison with the gold standard, there were 26 correct diagnoses by teleradiology (84%) and five incorrect (16%). Head CT scans accounted for 74% of the transmitted scans and all five of the cases with discrepant diagnoses. The cause of the single clinically relevant discrepancy was found to be unrelated to the use of teleradiology. In comparison with the formal report, the accuracy of the teleradiologist was not significantly different for all categories combined or for head CT scans alone. Video-capture technology provided a reliable means of transmitting CT scans for after-hours interpretation in emergency cases. PMID- 11027126 TI - A simple telemedicine system using a digital camera. AB - Radiographs on a viewing box were photographed at a remote hospital in South Africa using a digital camera with a resolution of 1024 x 768 pixels at 24-bit colour depth. The resultant images were stored in JPEG format and transmitted as email attachments to be read on a PC monitor by radiologists in Durban and Cape Town. Twenty-seven images were received, of which 23 were of diagnostic quality (85%). The mean file size was 120 kByte. For quality control purposes, 100 chest radiographs were photographed at a base hospital and read by a radiologist blinded to the diagnosis. In this study 96 images were of diagnostic quality (96%) and the correct diagnosis was made in 90 cases (94%). Incorrect readings were made in six cases (6%): small pulmonary nodules (less than 1 cm in diameter) were missed in five cases and in one case early apical tuberculosis was missed. Digital camera technology permits simple, inexpensive telemedicine. Limited spatial resolution is a concern when reading chest images with small pulmonary nodules and infiltrates. PMID- 11027127 TI - User satisfaction with teleconsultations for surgery. AB - We carried out a prospective study of realtime videoconferencing in surgical consultations. Videoconferencing equipment at the Satakunta Central Hospital, Pori, was connected by ISDN at 384 kbit/s to two health centres in the cities of Kankaanpaa and Huittinen (55 and 60 km from Pori, respectively). A document camera was used to transmit images of radiographs and paper documents. Fifty patients who needed a surgical consultation were examined by a doctor in the health centres, and the surgeon interviewed and observed the physical examination by videoconference. The consultation time ranged from 12 to 23 min (mean 15 min). Technically the equipment functioned reliably and the quality of the video picture was good. According to the consulting surgeon, the decision made in the videoconference was reliable in 48 cases (96%). According to the doctors in the health centres, the consultation was useful in 49 cases (98%) and was considered satisfactory in one. The doctors thought that the teleconsultation was as reliable as an outpatient appointment in 49 cases (98%). The educational benefit of the consultation was excellent or good in 38 cases (76%). The overall satisfaction of patients was very good or good in 45 cases (96%). All patients, except one, avoided travelling to a face-to-face appointment because they received a definite treatment decision during the teleconsultation. PMID- 11027128 TI - Videoconferencing for gynaecological cancer care: an international tumour board. PMID- 11027129 TI - A pilot study of telegenetics. PMID- 11027130 TI - Akt-dependent antiapoptotic action of insulin is sensitive to farnesyltransferase inhibitor. AB - CHO cells expressing the human insulin receptors (IR) were used to evaluate the effect of the potent farnesyltransferase inhibitor, manumycin, on insulin antiapoptotic function. Cell treatment with manumycin blocked insulin's ability to suppress pro-apoptotic caspase-3 activity which led to time-dependent proteolytic cleavage of two nuclear target proteins. The Raf-1/MEK/ERK cascade and the serine/threonine protein kinase Akt are two survival pathways that may be activated in response to insulin. We tested the hypothesis that inhibition of farnesylated Ras was causally related to manumycin-induced apoptosis and showed that the response to manumycin was found to be independent of K-Ras function because membrane association and activation of endogenous K-Ras proteins in terms of GTP loading and ERK activation were unabated following treatment with manumycin. Moreover, blocking p21Ras/Raf-1/MEK/ERK cascade by the expression of a transdominant inhibitory mSOS1 mutant in CHO-IR cells kept cells sensitive to the antiapoptotic action of insulin. Insulin-dependent activation of Akt was blocked by 4 h treatment with manumycin (P < 0.01), a kinetic too rapid to be explained by Ras inhibition. This study suggests that the depletion of short-lived farnesylated proteins by manumycin suppresses the antiapoptotic action of insulin at least in part by disrupting Akt activation but not that of the K-Ras/Raf-1/ERK dependent cascade. PMID- 11027131 TI - Impairment of the rod outer segment membrane guanylate cyclase dimerization in a cone-rod dystrophy results in defective calcium signaling. AB - Rod outer segment membrane guanylate cyclase1 (ROS-GC1) is the original member of the membrane guanylate cyclase subfamily whose distinctive feature is that it transduces diverse intracellularly generated Ca(2+) signals in the sensory neurons. In the vertebrate retinal neurons, ROS-GC1 is pivotal for the operations of phototransduction and, most likely, of the synaptic activity. The phototransduction- and the synapse-linked domains are separate, and they are located in the intracellular region of ROS-GC1. These domains sense Ca(2+) signals via Ca(2+)-binding proteins. These proteins are ROS-GC activating proteins, GCAPs. GCAPs control ROS-GC1 activity through two opposing regulatory modes. In one mode, at nanomolar concentrations of Ca(2+), the GCAPs activate the cyclase and as the Ca(2+) concentrations rise, the cyclase is progressively inhibited. This mode operates in phototransduction via two GCAPs: 1 and 2. The second mode occurs at micromolar concentrations of Ca(2+) via S100beta. Here, the rise of Ca(2+) concentrations progressively stimulates the enzyme. This mode is linked with the retinal synaptic activity. In both modes, the final step in Ca(2+) signal transduction involves ROS-GC dimerization, which causes the cyclase activation. The identity of the dimerization domain is not known. A heterozygous, triple mutation -E786D, R787C, T788M- in ROS-GC1 has been connected with autosomal cone-rod dystrophy in a British family. The present study shows the biochemical consequences of this mutation on the phototransduction- and the synapse-linked components of the cyclase. (1) It severely damages the intrinsic cyclase activity. (2) It significantly raises the GCAP1- and GCAP2-dependent maximal velocity of the cyclase, but this compensation, however, is not sufficient to override the basal cyclase activity. (3) It converts the cyclase into a form that only marginally responds to S100beta. The mutant produces insufficient amounts of the cyclic GMP needed to drive the machinery of phototransduction and of the retinal synapse at an optimum level. The underlying cause of the breakdown of both types of machinery is that, in contrast to the native ROS-GC1, the mutant cyclase is unable to change from its monomeric to the dimeric form, the form required for the functional integrity of the enzyme. The study defines the CORD in molecular terms, at a most basic level identifies a region that is critical in its dimer formation, and, thus, discloses a single unifying mechanistic theme underlying the complex pathology of the disease. PMID- 11027132 TI - Preparation, characterization, and the crystal structure of the inhibitor ZK 807834 (CI-1031) complexed with factor Xa. AB - Factor Xa plays a critical role in the formation of blood clots. This serine protease catalyzes the conversion of prothrombin to thrombin, the first joint step that links the intrinsic and extrinsic coagulation pathways. There is considerable interest in the development of factor Xa inhibitors for the intervention in thrombic diseases. This paper presents the structure of the inhibitor ZK-807834, also known as CI-1031, bound to factor Xa and provides the details of the protein purification and crystallization. Results from mass spectrometry indicate that the factor Xa underwent autolysis during crystallization and the first EGF-like domain was cleaved from the protein. The crystal structure of the complex shows that the amidine of ZK-807834 forms a salt bridge with Asp189 in the S1 pocket and the basic imidazoline fits snugly into the S4 site. The central pyridine ring provides a fairly rigid linker between these groups. This rigidity helps minimize entropic losses during binding. In addition, the structure reveals new interactions that were not found in the previous factor Xa/inhibitor complexes. ZK-807834 forms a strong hydrogen bond between an ionized 2-hydroxy group and Ser195 of factor Xa. There is also an aromatic ring-stacking interaction between the inhibitor and Trp215 in the S4 pocket. These interactions contribute to both the potency of this compound (K(I) = 0.11 nM) and the >2500-fold selectivity against homologous serine proteases such as trypsin. PMID- 11027133 TI - Crystal structure of human cathepsin V. AB - Cathepsin V is a lysosomal cysteine protease that is expressed in the thymus, testis and corneal epithelium. We have determined the 1.6 A resolution crystal structure of human cathepsin V associated with an irreversible vinyl sulfone inhibitor. The fold of this enzyme is similar to the fold adopted by other members of the papain superfamily of cysteine proteases. This study provides a framework for understanding the structural basis for cathepsin V's activity and will aid in the design of inhibitors of this enzyme. A comparison of cathepsin V's active site with the active sites of related proteases revealed a number of differences, especially in the S2 and S3 subsites, that could be exploited in identifying specific cathepsin V inhibitors or in identifying inhibitors of other cysteine proteases that would be selective against cathepsin V. PMID- 11027134 TI - Residue R216 and catalytic efficiency of a murine class alpha glutathione S transferase toward benzo[a]pyrene 7(R),8(S)-diol 9(S), 10(R)-epoxide. AB - Murine class alpha glutathione S-transferase A1-1 (mGSTA1-1), unlike mammalian class alpha GSTs, is the most efficient in the glutathione (GSH) conjugation of the ultimate carcinogenic metabolite of benzo[a]pyrene, (+)-anti-7,8-dihydroxy 9,10-oxy-7,8,9, 10-tetrahydrobenzo[a]pyrene [(+)-anti-BPDE] [Hu, X., Srivastava, S. K., Xia, H., Awasthi, Y. C., and Singh, S. V. (1996) J. Biol. Chem. 271, 32684 32688]. Here, we report the crystal structures of mGSTA1-1 in complex with GSH and with the GSH conjugate of (+)-anti-BPDE (GSBpd) at 1.9 and 2.0 A resolution, respectively. Both crystals belong to monoclinic space group C2 with one dimer in the asymmetric unit. The structures reveal that, within one subunit, the GSH moiety interacts with residues Y8, R14, K44, Q53, V54, Q66, and T67, whereas the hydrophobic moiety of GSBpd interacts with the side chains of F9, R14, M207, A215, R216, F219, and I221. In addition, the GSH moiety interacts with D100 and R130 from the other subunit across the dimer interface. The structural comparison between mGSTA1-1.GSH and mGSTA1-1.GSBpd reveals significant conformational differences. The movement of helix alpha9 brings the residues on the helix into direct interaction with the product. Most noticeable are the positional displacement and conformational change of R216, one of the residues located in helix alpha9. The side chain of R216, which points away from the H-site in the mGSTA1-1.GSH complex, probes into the active site and becomes parallel with the aromatic ring system of GSBpd. Moreover, the guanidinium group of R216 shifts approximately 8 A and forms a strong hydrogen bond with the C8 hydroxyl group of GSBpd, suggesting that the electrostatic assistance provided by the guanidinium group facilitates the ring-opening reaction of (+)-anti-BPDE. The structure of mGSTA1-1. GSBpd is also compared with those of hGSTP1-1[V104,A113].GSBpd, hGSPA1 1.S-benzylglutathione, and mGSTA4-4. 4-S-glutathionyl-5-pentyltetrahydrofuran-2 ol. The comparison provides further evidence that supports the functional roles of R216 and helix alpha9. The lack of mobility of helix alpha9 and/or the lack of electrostatic assistance from R216 may be responsible for the relatively lower activity of hGSTA1-1, mGSTA4-4, and hGSTP1-1 toward (+)-anti-BPDE. PMID- 11027135 TI - Conformational preferences of a synthetic 30mer peptide from the interface between the neck and stalk regions of kinesin. AB - The conformation of a synthetic peptide, consisting of 30 amino acids spanning the neck and hinge regions of rat brain kinesin, was investigated by NMR spectroscopy. The peptide extends from K357 to D386 and has the sequence KSVIQHLEVELNRWRNGEAVPEDEQISAKD. A total of 82 distance range constraints and 23 dihedral angle constraints could be obtained from NOESY and E.COSY spectra, respectively. These were used to calculate 500 structures by applying the REDAC algorithm of the software package DYANA. The first half of the peptide matched the helical structure of the neck determined from an X-ray crystal structure of kinesin. This part normally dimerizes into a coiled-coil by virtue of a leucine zipper interaction, but it is alpha-helical even in the monomeric state. The second half (not visible in the X-ray structure because of disorder) contains locally defined structure elements (extended chain, helical loop) connected by flexible joints. This is consistent with the "hinge" function postulated for this domain which is important for kinesin's motility and orientation. PMID- 11027136 TI - Direct photoaffinity labeling of cellular retinoic acid-binding protein I (CRABP I) with all-trans-retinoic acid: identification of amino acids in the ligand binding site. AB - Cellular retinoic acid-binding proteins I and II (CRABP-I and -II, respectively) are transport proteins for all-trans-retinoic acid (RA), an active metabolite of vitamin A (retinol), and have been reported to be directly involved in the metabolism of RA. In this study, direct photoaffinity labeling with [11,12 (3)H]RA was used to identify amino acids comprising the ligand binding site of CRABP-I. Photoaffinity labeling of CRABP-I with [(3)H]RA was light- and concentration-dependent and was protected by unlabeled RA and various retinoids, indicating that the labeling was directed to the RA-binding site. Photolabeled CRABP-I was hydrolyzed with endoproteinase Lys-C to yield radioactive peptides, which were separated by reversed-phase HPLC for analysis by Edman degradation peptide sequencing. This method identified five modified amino acids from five separate HPLC fractions: Trp7, Lys20, Arg29, Lys38, and Trp109. All five amino acids are located within one side of the "barrel" structure in the area indicated by the reported crystal structure as the ligand binding site. This is the first direct identification of specific amino acids in the RA-binding site of CRABPs by photoaffinity labeling. These results provide significant information about the ligand binding site of the CRABP-I molecule in solution. PMID- 11027137 TI - Hypermodified nucleosides in the anticodon of tRNALys stabilize a canonical U turn structure. AB - Modified nucleosides in the anticodon domain of Escherichia coli tRNA(Lys) are necessary for high-affinity codon recognition and reading frame maintenance. Human tRNA(Lys,3) is the specific primer for HIV-1 reverse transcriptase and also requires nucleoside modification for proper function. We now present NMR solution structures for the fully modified 17-nucleotide E. coli tRNA(Lys) anticodon stem loop domain (ASL). NMR data were also collected for several partially modified ASLs, revealing the contributions each modified nucleoside (mnm(5)s(2)U34, t(6)A37, and psi39) makes in transforming the disordered, unmodified tRNA ASL into the highly ordered native structure. The solution structure of the native ASL domain provides insight into longstanding questions regarding both wobble position modification and the nearly ubiquitous t(6)A37 found in tRNAs with an adjacent U at position 36. Native tRNA(Lys) has a U-turn structure similar to the yeast tRNA(Phe) crystal structure, unlike previously proposed "unconventional" anticodon structures characterized by stable interactions between mnm(5)s(2)U-34 and t(6)A-37. PMID- 11027138 TI - Stressing-out DNA? The contribution of serine-phosphodiester interactions in catalysis by uracil DNA glycosylase. AB - The DNA repair enzyme uracil DNA glycosylase (UDG) pinches the phosphodiester backbone of damaged DNA using the hydroxyl side chains of a conserved trio of serine residues, resulting in flipping of the deoxyuridine from the DNA helix into the enzyme active site. We have investigated the energetic role of these serine-phosphodiester interactions using the complementary approaches of crystallography, directed mutagenesis, and stereospecific phosphorothioate substitutions. A new crystal structure of UDG bound to 5'-HO-dUAAp-3' (which lacks the 5' phosphodiester group that interacts with the Ser88 pinching finger) shows that the glycosidic bond of dU has been cleaved, and that the enzyme has undergone the same specific clamping motion that brings key active site groups into position as previously observed in the structures of human UDG bound to large duplex DNA substrates. From this structure, it may be concluded that glycosidic bond cleavage and the induced fit conformational change in UDG can occur without the 5' pinching interaction. The S88A, S189A, and S192G "pinching" mutations exhibit 360-, 80-, and 21-fold damaging effects on k(cat)/K(m), respectively, while the S88A/S189A double mutant exhibits an 8200-fold damaging effect. A free energy analysis of the combined effects of nonbridging phosphorothioate substitution and mutation at these positions reveals the presence of a modest amount of strain energy between the compressed 5' and 3' phosphodiester groups flanking the bound uridine. Overall, these results indicate a role for these serine-phosphodiester interactions in uracil flipping and preorganization of the sugar ring into a reactive conformation. However, in contrast to a recent proposal [Parikh, S. S., et al. (2000) Proc Natl. Acad. Sci. 94, 5083], there is no evidence that conformational strain of the glycosidic bond induced by serine pinching plays a major role in the 10(12)-fold rate enhancement brought about by UDG. PMID- 11027139 TI - Interactions of the NADP(H)-binding domain III of proton-translocating transhydrogenase from escherichia coli with NADP(H) and the NAD(H)-binding domain I studied by NMR and site-directed mutagenesis. AB - Using the purified NADP(H)-binding domain of proton-translocating Escherichia coli transhydrogenase (ecIII) overexpressed in (15)N- and (2)H-labeled medium, together with the purified NAD(H)-binding domain from E. coli (ecI), the interface between ecIII and ecI, the NADP(H)-binding site and the influence on the interface by NAD(P)(H) was investigated in solution by NMR chemical shift mapping. Mapping of the NADP(H)-binding site showed that the NADP(H) substrate is bound to ecIII in an extended conformation at the C-terminal end of the parallel beta-sheet. The distribution of chemical shift perturbations in the NADP(H) binding site, and the nature of the interaction between ecI and ecIII, indicated that the nicotinamide moiety of NADP(H) is located near the loop comprising residues P346-G353, in agreement with the recently determined crystal structures of bovine [Prasad, G. S., et al. (1999) Nat. Struct. Biol. 6, 1126-1131] and human heart [White, A. W., et al. (2000) Structure 8, 1-12] transhydrogenases. Further chemical shift perturbation analysis also identified regions comprising residues G389-I406 and G430-V434 at the C-terminal end of ecIII's beta-sheet as part of the ecI-ecIII interface, which were regulated by the redox state of the NAD(P)(H) substrates. To investigate the role of these loop regions in the interaction with domain I, the single cysteine mutants T393C, R425C, G430C, and A432C were generated in ecIII and the transhydrogenase activities of the resulting mutant proteins characterized using the NAD(H)-binding domain I from Rhodospirillum rubrum (rrI). All mutants except R425C showed altered NADP(H) binding and domain interaction properties. In contrast, the R425C mutant showed almost exclusively changes in the NADP(H)-binding properties, without changing the affinity for rrI. Finally, by combining the above conclusions with information obtained by a further characterization of previously constructed mutants, the implications of the findings were considered in a mechanistic context. PMID- 11027140 TI - Structure of lithocholic acid binding to the N-terminal 8-kDa domain of DNA polymerase beta. AB - The purpose of this study was to investigate the molecular action of lithocholic acid (LCA), known as a selective inhibitor of DNA polymerase beta (pol beta). The 39-kDa pol beta was separated proteolytically into two fragments of the template primer binding domain (8 kDa) and the catalytic domain (31 kDa). LCA bound tightly to the 8-kDa fragment but not to the 31-kDa fragment. We examined the structural interaction with the 8-kDa domain using LCA. On (1)H-(15)N HMQC NMR analysis of pol beta with LCA, the 8-kDa domain bound to LCA as a 1:1 complex with a dissociation constant (K(D)) of 1.56 mM. The chemical shifts were observed only in residues mainly in helix-3, helix-4, and the 79-87 turn of the same face. No significant shifts were observed for helix-1, helix-2, and other loops of the 8-kDa domain. This region was composed mainly of three amino acid residues (Lys60, Leu77, and Thr79) of pol beta on the LCA interaction interface. The inhibition mechanism and the structure-function relationship between pol beta and LCA is discussed. PMID- 11027141 TI - Flexibility and ligand exchange in a buried cavity mutant of T4 lysozyme studied by multinuclear NMR. AB - The Leu99-->Ala mutant of T4 lysozyme contains a large internal cavity in the core of its C-terminal domain that is capable of reversibly binding small hydrophobic compounds. Although the cavity is completely buried, molecules such as benzene or xenon can exchange rapidly in and out. The dynamics of the unliganded protein have been compared to the wild-type protein by measuring the NMR spin relaxation rates of backbone amide and side chain methyl nuclei. Many residues surrounding the cavity were found to be affected by a chemical exchange process with a rate of 1500 +/- 200 s(-1), which is quenched upon addition of saturating amounts of the ligand xenon. The relationship between the structure, dynamics, and energetics of the T4 lysozyme mutant is discussed. PMID- 11027142 TI - Intestinal sterol absorption mediated by scavenger receptors is competitively inhibited by amphipathic peptides and proteins. AB - Exchangeable serum apolipoproteins and amphipathic alpha-helical peptides are effective inhibitors of sterol (free and esterified cholesterol) uptake at the small-intestinal brush border membrane. The minimal structural requirement of an inhibitor is an amphipathic alpha-helix of 18 amino acids. The inhibition is competitive, indicating that the inhibitor binds to scavenger receptor class B type I (SR-BI) present in the brush border membrane and responsible for sterol uptake. Binding of apolipoprotein A-I to SR-BI of rabbit brush border membrane is cooperative, characterized by a dissociation constant K(d) = 0.45 microM and a Hill coefficient of n = 2.8. The cooperativity of the interaction is due to binding of the inhibitor molecule to a dimeric or oligomeric form of SR-BI held together by disulfide bridges. Consistent with the competitive nature of the inhibition, the K(d) value agrees within experimental error with the IC(50) value of inhibition and with the inhibition constant K(I). After proteinase K treatment of brush border membrane vesicles, the affinity of the interaction of apolipoprotein A-I expressed as K(d) is reduced by a factor of 20, and the cooperativity is lost. The interaction of proteinase K-treated brush border membrane vesicles with apolipoprotein A-I is nonspecific partitioning of the apolipoprotein into the lipid bilayer of brush border membrane vesicles. PMID- 11027143 TI - Anion size modulates the structure of the A state of cytochrome c. AB - Several studies have shown that anions induce collapse of acid-denatured cytochrome c into the compact A state having the properties of the molten globule and that the anion charge is the main determinant for the A state stabilization. The results here reported show that the anion size plays a role in determining the overall structure of the A state. In particular, small anions induce formation of an A state in which the native Met80-Fe(III) axial bond is recovered and the nativelike redox properties restored. On the other hand, the A state stabilized by large anions shows a histidine (His26 or His33) as the sixth ligand of the heme-iron, a very weak interaction between Trp59 and the heme propionate, and lacks nativelike redox properties. The two anion-stabilized states show similar stability, indicating that (i) the hydrophobic core (which is equally stabilized by all the anions investigated, independently of their size) is the region that mainly contributes to the macromolecule stabilization, and (ii) the flexible loops are responsible for the spectroscopic (and, thus, structural) and redox differences observed. PMID- 11027144 TI - Conformational analysis of site-specific DNA cross-links of cisplatin-distamycin conjugates. AB - The requirement for novel platinum antitumor drugs led to the concept of synthesis of novel platinum drugs based on targeting cisplatin to various carrier molecules. We have shown [Loskotova, H., and Brabec, V. (1999) Eur. J. Biochem. 266, 392-402] that attachment of DNA minor-groove-binder distamycin to cisplatin changes several features of DNA-binding mode of the parent platinum drug. Major differences comprise different conformational changes in DNA and a considerably higher interstrand cross-linking efficiency. The studies of the present work have been directed to the analysis of oligodeoxyribonucleotide duplexes containing single, site-specific adducts of platinum-distamycin conjugates. These uniquely modified duplexes were analyzed by Maxam-Gilbert footprinting, phase-sensitive gel electrophoresis bending assay and chemical probes of DNA conformation. The results have indicated that the attachment of distamycin to cisplatin mainly affects the sites involved in the interstrand cross-links so that these adducts are preferentially formed between complementary guanine and cytosine residues. This interstrand cross-link bends the helix axis by approximately 35 degrees toward minor groove, unwinds DNA by approximately 95 degrees and distorts DNA symmetrically around the adduct. In addition, CD spectra of restriction fragments modified by the cisplatin-distamycin conjugates have demonstrated that distamycin moiety in the interstrand cross-links of these compounds interacts with DNA. This interaction facilitates the formation of these adducts. Hence, the structural impact of the specific interstrand cross-link detected in this study deserves attention when biological behavior of cisplatin derivatives targeted by oligopeptide DNA minor-groove-binders is evaluated. PMID- 11027145 TI - Parallel self-associated structures formed by T,C-rich sequences at acidic pH. AB - Oligonucleotides of nonregular heteropyrimidine sequences incorporating or not incorporating purine residues 5'-d(ACTCCCTTCTCCTCTCTA), 5'-d(ACTCCCTGGTCCTCTCTA), 5'-d(TCTCTCCTGGTCCCTCC), and 5'-d(TCTCTCCTCTTCCCTCC) can form self-associated parallel-stranded (ps) structures at pH 4-5.5. The ps structures were identified by studying at neutral and acidic pH UV melting transitions, FTIR spectra, and fluorescence of pyrene-labeled oligonucleotides as well as by chemical joining of 5'-phosphorylated oligonucleotides. A gel electrophoresis run for oligonucleotides 5'-d(TCTCTCCTCTTCCCTCC) and 5'-d(ACTCCCTTCTCCTCTCTA) has shown the formation of homoduplexes at low DNA strand concentrations. Ps structures are held by C-C(+) base pairs and have N- and S-types of sugar puckering as detected by FTIR spectroscopy in the millimolar concentration range. Guanine inserts as well as thymine and purine inserts into an oligomeric cytosine sequence make the formation of the tetraplex i-motif unfavorable. MvaI restriction endonuclease, which recognizes the CCT/AGG sequence in DNA, does not cleave parallel pseudosubstrates. PMID- 11027146 TI - Influence of alpha-CH-->NH substitution in C8-CoA on the kinetics of association and dissociation of ligands with medium chain acyl-CoA dehydrogenase. AB - We previously reported that the kinetic profiles for the association and dissociation of functionally diverse C(8)-CoA-ligands, viz., octanoyl-CoA (substrate), octenoyl-CoA (product), and octynoyl-CoA (inactivator) with medium chain acyl-CoA dehydrogenase (MCAD), were essentially identical, suggesting that the protein conformational changes played an essential role during ligand binding and/or catalysis [Peterson, K. L., Sergienko, E. E., Wu, Y., Kumar, N. R., Strauss, A. W., Oleson, A. E., Muhonen, W. W., Shabb, J. B., and Srivastava, D. K. (1995) Biochemisry 34, 14942-14953]. To ascertain the structural basis of the above similarity, we investigated the kinetics of association and dissociation of alpha-CH-->NH-substituted C(8)-CoA, namely, 2-azaoctanoyl-CoA, with the recombinant form of human liver MCAD. The rapid-scanning and single wavelength stopped-flow data for the binding of 2-azaoctanoyl-CoA to MCAD revealed that the overall interaction proceeds via two steps. The first (fast) step involves the formation of an enzyme-ligand collision complex (with a dissociation constant of K(c)), followed by a slow isomerization step (with forward and reverse rate constants of k(f) and k(r), respectively) with concomitant changes in the electronic structure of the enzyme-bound FAD. Since the latter step involves a concurrent change in the enzyme's tryptophan fluorescence, it is suggested that the isomerization step is coupled to the changes in the protein conformation. Although the overall binding affinity (K(d)) of the enzyme-2-azaoctanoyl-CoA complex is similar to that of the enzyme-octenoyl-CoA complex, their microscopic equilibria within the collision and isomerized complexes show an opposite relationship. These results coupled with the isothermal titration microcalorimetric studies lead to the suggestion that the electrostatic interaction within the enzyme site phase modulates the microscopic steps, as well as their corresponding ground and transition states, during the course of the enzyme-ligand interaction. PMID- 11027147 TI - Determination of the pKa value of C115 in MurA (UDP-N-acetylglucosamine enolpyruvyltransferase) from Enterobacter cloacae. AB - The enzyme UDP-N-acetylglucosamine (UDP-NAG) enolpyruvyltransferase (MurA) catalyzes the formation of enolpyruvyl-UDP-NAG, a precursor in peptidoglycan biosynthesis. The residue at position 115 in MurA has been proposed to act as a general acid in the enzymatic reaction. This is also the primary site of action of the antibiotic fosfomycin. In this paper, the pK(a) of Cys-115 has been determined to be 8.3, by titration of Enterobacter cloacae MurA with the alkylating agent iodoacetamide as a function of pH. Use of site-directed mutagenesis has established that only C115 is essential for catalysis, and the three other cysteine residues (C251, C354, and C381) are nonessential. Mass spectrometric analysis demonstrated that C115 is not alkylated at pH <7, but is alkylated significantly at pH >7. Measurement of the enzymatic inhibition by iodoacetamide as a function of pH showed maximum inhibition at pH >9, with a second-order rate constant of inhibition of 44 M(-)(1) s(-)(1) at pH 10. The presence of either one of the substrates did not influence the inactivation behavior, while the presence of both substrates resulted in a 5-fold reduction in the extent of alkylation. The covalent species that results from PEP bound to C115 of MurA exhibited 50-100-fold increased resistance against alkylation by iodoacetamide. These results imply that C115 is appreciably protonated at physiological pH and, therefore, is capable of acting as a proton donor in the enzyme-catalyzed reaction. However, it also implies that C115 is appreciably deprotonated at physiological pH also, whereupon the resultant thiolate nucleophile may play an important role in the formation of the covalent O phosphothioketal species, whose role in catalysis is yet to be established. PMID- 11027148 TI - Energetic consequences of accommodating a bulkier ligand at the active site of medium chain acyl-CoA dehydrogenase by creating a complementary enzyme site cavity. AB - The substitution of the C=O by the C=S group in 2-azaoctanoyl-CoA increases the volume of the ligand by 11 A(3), and the excision of a methylene group from Glu 376, via Glu-376 --> Asp (E376D) mutation in medium chain acyl-CoA dehydrogenase (MCAD), creates a complementary cavity of 18 A(3) dimension, just opposite to the ligand's carbonyl group. We investigated whether the newly created cavity would facilitate accommodation of the bulkier (C=O --> C=S substituted) ligand within the active site of the enzyme. To ascertain this, we determined the binding affinity and kinetics of association and dissociation of 2-azaoctanoyl-CoA and the C=O --> C=S substituted ligand, 2-azadithiooctanoyl-CoA, involving the wild type and Glu-376 --> Asp mutant enzymes. The experimental data revealed that the binding of 2-azadithiooctanoyl-CoA to the wild-type enzyme was energetically unfavorable as compared to 2-azaoctanoyl-CoA. However, such an energetic constraint was alleviated for the binding of the former ligand to the E376D mutant enzyme site. A detailed account of the free energy and enthalpic profiles for the binding of 2-azaoctanoyl-CoA and 2-azadithiooctanoyl-CoA to the wild-type and Glu-376 --> Asp mutant enzymes throws light on the flexibility of the enzyme site cavity in stabilizing the ground and transition states of the enzyme-ligand complexes. PMID- 11027149 TI - Intersubunit interactions in human X,K-ATPases: role of membrane domains M9 and M10 in the assembly process and association efficiency of human, nongastric H,K ATPase alpha subunits (ATP1al1) with known beta subunits. AB - Na,K- and H,K-ATPase (X,K-ATPase) alpha subunits need association with a beta subunit for their maturation, but the authentic beta subunit of nongastric H,K ATPase alpha subunits has not been identified. To better define alpha-beta interactions in these ATPases, we coexpressed human, nongastric H,K-ATPase alpha (AL1) and Na,K-ATPase alpha1 (alpha1NK) as well as AL1-alpha1 and alpha1-AL1 chimeras, which contain exchanged M9 and M10 membrane domains, together with each of the known beta subunits in Xenopus oocytes and followed their resistance to cellular and proteolytic degradation and their ER exit. We show that all beta subunits (gastric betaHK, beta1NK, beta2NK, beta3NK, or Bufo bladder beta) can associate efficiently with alpha1NK, but only gastric betaHK, beta2NK, and Bufo bladder beta can form stably expressed AL1-beta complexes that can leave the ER. The trypsin resistance and the forces of subunit interaction, probed by detergent resistance, are lower for AL1-beta complexes than for alpha1NK-beta complexes. Furthermore, chimeric alpha1-AL1 can be stabilized by beta subunits, but alpha1 AL1-gastric betaHK complexes are retained in the ER. On the other hand, chimeric AL1-alpha1 cannot be stabilized by any beta subunit. In conclusion, these results indicate that (1) none of the known beta subunits is the real partner subunit of AL1 but an as yet unidentified, authentic beta should have structural features resembling gastric betaHK, beta2NK, or Bufo bladder beta and (2) beta-mediated maturation of alpha subunits is a multistep process which depends on the membrane insertion properties of alpha subunits as well as on several discrete events of intersubunit interactions. PMID- 11027150 TI - Functional interactions in cytochrome P450BM3. Evidence that NADP(H) binding controls redox potentials of the flavin cofactors. AB - NADP(H) binding is essential for fast electron transfer through the flavoprotein domain of the fusion protein P450BM3. Here we characterize the interaction of NADP(H) with the oxidized and partially reduced enzyme and the effect of this interaction on the redox properties of flavin cofactors and electron transfer. Measurements by three different approaches demonstrated a relatively low affinity of oxidized P450BM3 for NADP(+), with a K(d) of about 10 microM. NADPH binding is also relatively weak (K(d) approximately 10 microM), but the affinity increases manyfold upon hydride ion transfer so that the active 2-electron reduced enzyme binds NADP(+) with a K(d) in the submicromolar range. NADP(H) binding induces conformational changes of the protein as demonstrated by tryptophan fluorescence quenching. Fluorescence quenching indicated preferential binding of NADPH by oxidized P450BM3, while no catalytically competent binding with reduced P450BM3 could be detected. The hydride ion transfer step, as well as the interflavin electron transfer steps, is readily reversible, as demonstrated by a hydride ion exchange (transhydrogenase) reaction between NADPH and NADP(+) or their analogues. Experiments with FMN-free mutants demonstrated that FAD is the only flavin cofactor required for the transhydrogenase activity. The equilibrium constants of each electron transfer step of the flavoprotein domain during catalytic turnover have been calculated. The values obtained differ from those calculated from equilibrium redox potentials by as much as 2 orders of magnitude. The differences result from the enzyme's interaction with NADP(H). PMID- 11027151 TI - Transmembrane peptide NB of influenza B: a simulation, structure, and conductance study. AB - The putative transmembrane segment of the ion channel forming peptide NB from influenza B was synthesized by standard solid-phase peptide synthesis. Insertion into the planar lipid bilayer revealed ion channel activity with conductance levels of 20, 61, 107, and 142 pS in a 0.5 M KCl buffer solution. In addition, levels at -100 mV show conductances of 251 and 413 pS. A linear current-voltage relation reveals a voltage-independent channel formation. In methanol and in vesicles the peptide appears to adopt an alpha-helical-like structure. Computational models of alpha-helix bundles using N = 4, 5, and 6 NB peptides per bundle revealed water-filled pores after 1 ns of MD simulation in a solvated lipid bilayer. Calculated conductance values [using HOLE (Smart et al. (1997) Biophys. J. 72, 1109-1126)] of ca. 20, 60, and 90 pS, respectively, suggested that the multiple conductance levels seen experimentally must correspond to different degrees of oligomerization of the peptide to form channels. PMID- 11027152 TI - Chain length determination of prenyltransferases: both heteromeric subunits of medium-chain (E)-prenyl diphosphate synthase are involved in the product chain length determination. AB - Among prenyltransferases, medium-chain (E)-prenyl diphosphate synthases are unusual because of their heterodimeric structures. The larger subunit has highly conserved regions typical of (E)-prenyltransferases. The smaller one has recently been shown to be involved in the binding of allylic substrate as well as determining the chain length of the reaction product [Zhang, Y.-W., et al. (1999) Biochemistry 38, 14638-14643]. To better understand the product chain length determination mechanism of these enzymes, several amino acid residues in the larger subunits of Micrococcus luteus B-P 26 hexaprenyl diphosphate synthase and Bacillus subtilis heptaprenyl diphosphate synthase were selected for substitutions by site-directed mutagenesis and examined by combination with the corresponding wild-type or mutated smaller subunits. Replacement of the Ala at the fifth position upstream to the first Asp-rich motif with bulky amino acids in both larger subunits resulted in shortening the chain lengths of the major products, and a double combination of mutant subunits of the heptaprenyl diphosphate synthase, I-D97A/II-A79F, yielded exclusively geranylgeranyl diphosphate. However, the combination of a mutant subunit and the wild-type, I Y103S/II-WT or I-WT/II-I76G, produced a C(40) prenyl diphosphate, and the double combination of the mutants, I-Y103S/II-I76G, gave a reaction product with longer prenyl chain up to C(50). These results suggest that medium-chain (E)-prenyl diphosphate synthases take a novel mode for the product chain length determination, in which both subunits cooperatively participate in maintaining and determining the product specificity of each enzyme. PMID- 11027153 TI - Pseudouridine synthase 3 from mouse modifies the anticodon loop of tRNA. AB - A cDNA encoding mouse pseudouridine synthase 3 (mPus3p) has been cloned. The predicted protein has 34% identity with yeast pseudouridine synthase 3 (Pus3), an enzyme known to form pseudouridine at positions 38 and 39 in yeast tRNA. The cDNA is 1.7 kb, and when used as a probe on a Northern blot of total RNA from mouse tissues or cells in culture, a band at 1.8 kb was observed. The open reading frame codes for a protein of 481 amino acids with a predicted molecular mass of 55 552 Da. When mPus3p was in vitro translated and used in reactions with tRNA substrates from both yeast and humans, uridines at position 39 were modified to pseudouridine. In a tRNA substrate with a uridine at position 38 (human tRNA(Leu)), there was very slight formation of pseudouridine at that position after incubation with mPus3p. PMID- 11027154 TI - Energetics of the induced structural change in a Ca2+ regulatory protein: Ca2+ and troponin I peptide binding to the E41A mutant of the N-domain of skeletal troponin C. AB - Structural studies have shown that the regulatory domains of skeletal and cardiac troponin C (sNTnC and cNTnC) undergo different conformational changes upon Ca(2+) binding; sNTnC "opens" with a large exposure of the hydrophobic surface, while cNTnC retains a "closed" conformation similar to that in the apo state. This is mainly due to the fact that there is a defunct Ca(2+)-binding site I in cNTnC. Despite the striking difference, the two proteins bind their respective troponin I (TnI) regions (sTnI(115-131) and cTnI(147-163), respectively) in a similar open fashion. Thus, there must exist a delicate energetic balance between Ca(2+) and TnI binding and the accompanying conformational changes in TnC for each system. To understand the coupling between Ca(2+) and TnI binding and the concomitant structural changes, we have previously engineered an E41A mutant of sNTnC and demonstrated that this mutation drastically reduced the Ca(2+)-binding affinity of site I in sNTnC, and as a result, E41A-sNTnC remains closed in the Ca(2+) bound state. In the present work, we investigated the interaction of E41A-sNTnC with the sTnI(115-131) peptide and found that the peptide binds to the Ca(2+) saturated E41A-sNTnC with a 1:1 stoichiometry and a dissociation constant of 300 +/- 100 microM. The peptide-induced chemical shift changes resemble those of Ca(2+) binding to sNTnC, suggesting that sTnI(115-131) induces the "opening" of E41A-sNTnC. In addition, the binding of sTnI(115-131) appears to be accompanied by a conformational change in site I of E41A-sNTnC so that the damaged regulatory site can bind Ca(2+) more tightly. Without Ca(2+), sTnI(115-131) only interacts with E41A-sNTnC nonspecifically. When Ca(2+) is titrated into E41A-sNTnC in the presence of sTnI(115-131), the Ca(2+)-binding affinity of site I was enhanced by approximately 5-fold as compared to when sTnI(115-131) was not present. These observations suggest that the binding of Ca(2+) and TnI is intimately coupled to each other. Together with our previous studies on Ca(2+) and TnI peptide binding to sNTnC and cNTnC, these results allow us to dissect the mechanism and energetics of coupling of ligand binding and structural opening intricately involved in the regulation of skeletal and cardiac muscle contraction. PMID- 11027155 TI - Controlling beta-sheet assembly in genetically engineered silk by enzymatic phosphorylation/dephosphorylation. AB - Enzymatic phosphorylation and dephosphorylation reactions were used to modify a genetically engineered variant of spider dragline silk. The approximately 25 kDa protein was phosphorylated with cyclic AMP-dependent kinase and dephosphorylated with calf intestinal alkaline phosphatase. Phosphorylation inhibited beta-sheet assembly of the protein and enhanced solubility to about 5 mg/mL in water, compared to about 20% of this level upon enzymatic dephosphorylation. The cyclability of the phosphorylation-dephosphorylation system was confirmed by MALDI with a model peptide. Kinetic studies conducted with [gamma-(32)P]ATP illustrate that the phosphorylation reaction proceeds over 6 h. Secondary structure of the phosphorylated and dephosphorylated proteins was determined by CD and FTIR. The results illustrate that an enzymatic phosphorylation event can be used to control the solution structure of a protein like silk, which has a tendency to prematurely precipitate due to the formation of beta-sheets. PMID- 11027156 TI - Growth of NS0 cells in protein-free, chemically defined medium. AB - Many hybridoma and recombinant myeloma cell lines have been successfully adapted to growth in protein-free media. Compared with serum-supplemented media, use of protein-free media promotes superior cell growth and protein expression and facilitates downstream purification of the expressed product. Owing to its sterol auxotrophy, the NS0 myeloma is normally grown in either a serum-supplemented medium or a serum-free medium supplemented with an animal-derived lipoprotein. CD Hybridoma Medium (a protein-free, chemically defined formulation) grows many cell lines that do not exhibit lipid dependence, but this medium does not support growth of NS0 cells without further lipid supplementation. We tested several commercially available lipid supplements in CD Hybridoma Medium, including bovine EX-CYTE VLE. None of the tested supplements supported long-term growth of NS0 cells in CD Hybridoma Medium. Sustained long-term growth of NS0 cells was achieved in CD Hybridoma Medium supplemented with various animal- or plant derived lipids complexed with cyclodextrin. NS0 cells adapted to CD Hybridoma Medium supplemented with cyclodextrin-lipid complex reached peak cell densities that were more than double those observed in serum-supplemented medium and were cultured for more than 15 passages. These cultures were also successfully cryopreserved and recovered in this defined medium. Through the use of cyclodextrin-based additives to CD Hybridoma Medium, it is possible to solubilize significant quantities of sterols and other lipids and to maintain a protein free, chemically defined cultivation environment for NS0 cells. The culture system can be kept entirely free of animal-derived components if the supplement is made with plant-derived or synthetic lipids. PMID- 11027157 TI - Plant protein hydrolysates: preparation of defined peptide fractions promoting growth and production in animal cells cultures. AB - A new approach was applied with the aim at producing plant protein hydrolysates less heterogeneous and less contaminated with nonpeptide substances than are the presently available digests. A significant reduction of nonprotein contaminants was achieved by extraction of the plant material, soy flour or wheat flour, with acetone prior to isolation of the protein. Enzymes of nonanimal origin, papain or Pronase, were used for protein hydrolysis. The components of the hydrolysates were resolved by low-pressure liquid chromatography. Separation of peptide fractions and of remaining nonpeptide contaminants was achieved using small-pore size-exclusion chromatography matrices, Sephadex G-15 or Biogel P-2. Individual peptide fractions, both from soy protein and from wheat gluten, varied substantially in their growth-promoting and production-enhancing activities when tested on a mouse hybridoma culture in protein-free medium. The highest enhancement of viable cell density in batch cultures was 180% of control, and the highest enhancement of final immunoglobulin concentration was more than 230% of control. The existence of marked differences in activity of individual peptide fractions leads to a suggestion that the hydrolysates may provide peptides exerting specific positive effects on cultured animal cells. PMID- 11027158 TI - Effects of insulin and LongR(3) on serum-free Chinese hamster ovary cell cultures expressing two recombinant proteins. AB - Insulin is the most commonly used growth factor for sustaining cell growth and viability in serum-free Chinese hamster ovary (CHO) cell cultures. In the present study insulin and IGF-1 analogue (LongR(3)) were compared for their ability to support growth, viability, and production of two serum-free CHO cell lines expressing recombinant protein. The first cell line, VA12, expresses protein B, and the second cell line, CL23, expresses protein C. Both molecules are recombinant cytokine receptors. VA12 will grow in serum-free media lacking growth factor, while CL23 requires either insulin or LongR(3) for cell growth. Both cell lines, however, require a growth factor for optimal performance under production conditions. In this study, LongR(3) was better able to sustain the viability of both cell lines under production conditions than insulin. These data indicate that while insulin and LongR(3) can both serve as growth and viability factors for CHO cells, LongR(3) is the preferred growth factor for cell lines VA12 and CL23. PMID- 11027159 TI - Insulin-like growth factor-I and transferrin mediate growth and survival of Chinese hamster ovary cells. AB - The aim of this investigation was to elucidate the roles of insulin-like growth factor-I (IGF-I) and transferrin in the survival and proliferation of Chinese hamster ovary (CHO) cells upon withdrawal of serum. For this purpose, we employed DNA analysis and flow cytometry to compare CHO cell lines expressing either IGF-I alone or IGF-I and transferrin. The ability of cells to cycle and the occurrence of apoptosis were monitored in these cells in serum-free medium. These results indicate that IGF-I alone is able to maintain the viability of CHO cells for an extended length of time in the absence of serum. Transferrin alone does not promote survival or proliferation. Only in the presence of both IGF-I and transferrin do cells survive and proliferate. Therefore, in attached CHO cultures, IGF-I alone does not stimulate cell proliferation but is a requirement for growth in serum-free medium in cooperation with transferrin. We report on the dual role of IGF-I as a survival factor in CHO cells and its interlocking role with transferrin to stimulate cell growth. PMID- 11027160 TI - Development of serum-free bioreactor production of recombinant human thyroid stimulating hormone receptor. AB - For the detection of autoantibodies to thyroid stimulating hormone receptors (TSH R) in Graves' disease based on a novel coated tube assay system, human TSH-R is needed in large amounts. Whereas expression of TSH-R in bacteria, yeast, or insect cells results in nonfunctional, denaturated receptor, mammalian cells such as COS, CHO, and HeLa are able to express functional TSH-R, but only in very low amounts. Furthermore, for all of these cultivations expensive standard media containing 10% fetal calf serum are needed to obtain functional receptor. Here we report on the development of a serum-free production-scale process based on a stable transformed and highly productive human leukemia cell line K562 (1). Starting with K562-TSH-R cells growing in medium containing 10% fetal calf serum the cell line was adapted to serum-free medium. The adaptation medium was optimized in regards to amino acid and protein concentrations, since the use of unadjusted medium caused cell death after 2 days. The adapted cells were stable and could be cultivated without antibiotics for more than 50 cell doublings without losing their productivity. The obtained receptor showed improved TSH binding. The process development was based on cultivations in a 2-L bench-scale bioreactor. Cultivations in batch mode and chemostat mode and perfusion cultivation with the usage of an internal microfiltration device and a spin filter device were compared. After process optimization a continuous process using spin-filter was set up and run in a 20 L-pilot-scale bioreactor. The presented results were the prerequisite for the production of the novel assay for the diagnosis of autoantibodies to TSH-R in Graves' disease. PMID- 11027161 TI - Amplified gene location in chromosomal DNA affected recombinant protein production and stability of amplified genes. AB - Previously, we established an easy and quick construction method for obtaining a stable and highly productive gene-amplified recombinant Chinese hamster ovary (CHO) cell line. With a gradual increase in methotrexate (MTX) concentration, gene-amplified cell pools had high and stable specific growth and production rates. Moreover, the phenotype of gene-amplified cells seemed to be affected by the location of the amplified gene in chromosomal DNA. We suspected that various kinds of gene-amplified cells might appear during the long-term selection to construct gene-amplified cell pools. To clarify the behavior of gene-amplified cell pools during a stepwise increase of MTX concentration, we isolated gene amplified clones derived from gene-amplified cell pools. We compared the characteristics of isolated clones, such as the productivity of recombinant protein, stability of amplified genes, and the location of amplified genes. As a result, telomere-type clones, in which the amplified gene was located near the telomeric region, were found to be more stable and productive than other types of clones. Telomere-type clones had over 100 copies of amplified genes in the chromosomal DNA. In contrast, a large number of other types of clones had less than 10 copies of amplified genes. During long-term cultivation in the absence of MTX, in other types of clones, amplified genes rapidly decreased in the chromosomal DNA. PMID- 11027162 TI - Investigating the secretory pathway of the baculovirus-insect cell system using a secretory green fluorescent protein. AB - The secretory pathway is important in actively transporting proteins into the extracellular environment of eucaryotic cells. In this study a green fluorescent protein (GFP) mutant engineered to contain a secretion signal was used as a model protein in order to visualize the secretion process inside insect cells. Fluorescent microscopy indicated that significant amounts of secreted green fluorescent protein (sGFP) accumulated in High-Five, Trichoplusia ni, cells following infection with a baculovirus vector containing the gene under the polyhedrin promoter. Laser scanning confocal microscopy was used to reconstruct whole cell images of the infected High-Five cells at multiple days postinfection. While the protein was widely distributed at 2 days postinfection, certain intracellular regions appeared to contain higher or lower concentrations of the sGFP. A layer by layer examination indicated pockets in which sGFP was absent, and these appear to be vesicles that have recently released the sGFP or are not yet accumulating sGFP. By 3 days postinfection, the sGFP in some cells was concentrated in a number of widely dispersed globules, which may represent the vesicle remnants of a deteriorating secretory pathway. In contrast, nonsecreted GFP was more uniformly distributed in the cells than sGFP and did not accumulate in vesicles. In addition to GFP, the lectins wheat germ agglutinin (WGA) and concanavalin A (ConA), which have affinities for sugar residues, were used to examine the secretory pathway. The WGA, which is a Golgi marker, was distributed around the nucleus prior to infection but then was found to be polarized in one region of the cell following the baculovirus infection. The expansion of other cellular compartments following the baculovirus infection may have caused a change in intracellular distribution of the Golgi. While some of the sGFP was found to colocalize with the WGA label, much of the sGFP was outside this Golgi region. In contrast, ConA labeling, which was not as specific as WGA, was found throughout the cell both before and after infection similar to the sGFP distribution. These studies demonstrate that confocal visualization of fluorescent proteins can be used as an in vivo tool for examining secretory processing in insect cells. PMID- 11027163 TI - An update of pTRIDENT multicistronic expression vectors: pTRIDENTs containing novel streptogramin-responsive promoters. AB - We present an update on the pTRIDENT multicistronic mammalian expression vectors and their implications in various metabolic engineering and therapeutic applications. The pTRIDENT vector family has been expanded by construction of a new set of pTRIDENT-based vectors containing constitutive promoters of human origin (ubiquitin C and EF-1alpha promoters) and selectable markers (zeocin resistance) and expressing different reporter genes (secreted alkaline phosphatase (SEAP) and the secreted single-chain urokinase-type plasminogen activator (low-M(r) u-PA)). In addition, we have constructed pTRIDENT derivatives with novel streptogramin-repressible and streptogramin-inducible promoters for simultaneous and adjustable expression of three different transgenes. Streptogramin-inducible and tetracycline-repressible pTRIDENT derivatives were used to simultaneously control expression of three fluorescent proteins in mammalian cells: the enhanced cyan fluorescent protein (CFP), the recently isolated red fluorescent protein (RFP, also designated dsRed), and the enhanced yellow fluorescent protein (YFP). Owing to their modular structure, the pTRIDENT vector family represents a construction kit for the design of novel multicistronic expression constructs. PMID- 11027164 TI - Effect of PDI overexpression on recombinant protein secretion in CHO cells. AB - In eukaryotic cells, protein disulfide isomerase (PDI) found in the endoplasmic reticulum (ER) catalyzes disulfide bond exchange and assists in protein folding of newly synthesized proteins. PDI also functions as a molecular chaperone and has been found associated with proteins in the ER. In addition, PDI functions as a subunit of two more complex enzyme systems: the prolyl-4-hydroxylase and the triacylglycerol transfer proteins. Increasing PDI activity in bacterial, yeast, and insect cell expression systems can lead to increased secretion of heterologous proteins containing disulfide bridges. Since Chinese hamster ovary (CHO) cells are widely used for the expression of recombinant proteins, we expressed recombinant human PDI (rhu PDI) in CHO cells to increase cellular PDI levels and examined its effect on the secretion of two different recombinant proteins: interleukin 15 (IL-15) and a tumor necrosis factor receptor:Fc fusion protein (TNFR:Fc). Secretion of TNFR:Fc (a disulfide-rich protein) is decreased in cells overexpressing PDI; the TNFR:Fc protein is retained inside these cells and colocalizes with the overexpressed rhu PDI protein in the endoplasmic reticulum. PDI overexpression did not result in intracellular retention of IL15. The nature of the interaction between PDI and TNFR:Fc was further investigated by expressing a disulfide isomerase mutant PDI in CHO cells to determine if the functional activity of PDI is involved in the cellular retention of TNFR:Fc protein. PMID- 11027165 TI - Production of HIV-1 gp120 in packed-bed bioreactor using the vaccinia virus/T7 expression system. AB - The HeLa cell-vaccinia virus system is an attractive method for producing recombinant mammalian proteins with proper post-translation modifications. This approach is especially important for the production of HIV-1 envelope glycoprotein, gp120, since more than half of its total mass is due to carbohydrates. A recombinant vaccinia virus/T7 RNA polymerase expression system was developed to express and produce large amounts of gp120 tagged with six histidine residues. In this system, the expressed T7 RNA polymerase from one virus drives the transcription of the gp120 encoded in the second virus. During the process development phase, the following parameters were studied: infection time, infection duration, multiplicity of infection, ratio of the two viruses, medium composition, and medium replacement strategy during the infection phase. The chosen production method was based on using the packed-bed bioreactor. The HeLa cells were immobilized on fibrous disks (Fibra-Cel) packed in an internal basket positioned in a vertically mixed bioreactor (Celligen Plus), and 25 g of carriers were packed in a 1.6-L (working volume) reactor. The process included a growth stage followed by a production stage. In the growth stage, the bed was perfused with a serum-containing medium, allowing the cells to grow to saturation, and in the production stage, done using serum-free medium, the cells were infected with the two recombinant viruses. The expressed protein was secreted, collected from the culture fluid, and purified. The specific production was found to be between 2 and 3 microg of protein/10(6) cells, and the volumetric production was around 10 mg/50 g carriers. PMID- 11027166 TI - Effect of ammonia on the glycosylation of human recombinant erythropoietin in culture. AB - Recombinant human erythropoietin (EPO) was produced by a stable transfected CHO K1 cell clone (EPO-81) grown in serum-free medium. Our previous work showed that there was a significant increase in the heterogeneity of the glycoforms of EPO and a reduction of the sialylation at 20 mM NH(4)Cl. In the work presented here, the effects of ammonia on EPO N-linked oligosaccharides were analyzed. EPO was purified from culture supernatants by immunoaffinity chromatography. The N-linked oligosaccharides were released enzymatically and analyzed by fluorophore-assisted carbohydrate electrophoresis (FACE) and HPLC. The FACE N-linked oligosaccharide profile showed that the sialylated glycans contain one prominent band at a position corresponding to eight glucose units. The density of the major band was greatly diminished and the width was significantly increased in cultures containing added ammonia. The proportion of tetraantennary structures was reduced by 60%, while the tri- and biantennary structures were increased proportionally in the presence of ammonia. Glycan analysis by HPLC using a weak anion exchange column showed that the most significant characteristic effect of ammonia was a reduction of the proportion of glycans with four sialic acids from 46% in control cultures to 29% in ammonia-treated cultures. Analysis of the desialylated glycans by normal phase chromatography indicated a distribution of tetra-, tri-, and biantennary structures similar to that shown by FACE. The N-linked glycan sequence was determined by sequential exoglycosidase digestion followed by FACE. The results indicated a typical N-linked complex oligosaccharide structure. Glycans from ammonia-containing cultures showed the same sequence pattern. In conclusion, we showed that ammonia in the culture medium affected EPO glycosylation, which was observed as a reduction of the tetraantennary and tetrasialylated oligosaccharide structures. However, the presence of ammonia in the cultures did not change the oligosaccharide sequence. PMID- 11027167 TI - Ammonia removal using hepatoma cells in mammalian cell cultures. AB - It was examined whether hepatocyte cell lines can be used for ammonia removal in mammalian cell cultures. It was found that there exists a critical ammonium concentration level for each hepatocyte cell to remove ammonia. Among the cells tested in this work, primary hepatocytes showed the strongest ammonia removal capability if ammonium concentration is higher than the critical level. However, primary hepatocytes lost the liver function gradually and finally died after 2-3 weeks. Because of this limitation, primary hepatocytes were not appropriate to be used for ammonia removal in long-term cultures. Hep G2 cells, which are immortal, also showed a strong ammonia removal activity. The ammonia removal activity of Hep G2 cells depended on the concentration of ammonium in the medium, as in the case of primary hepatocytes. However, urea could not be detected in the course of ammonia removal by Hep G2 cells. Instead of urea, Hep G2 cells secreted glutamine into the culture medium. The capacity for ammonia removal was higher in the absence than in the presence of glutamine. Thus we checked the activity of glutamine synthetase in the Hep G2 cells. The level of glutamine synthetase activity increased with the addition of ammonium chloride. This result accounts for the ammonium concentration dependency of Hep G2 cells in ammonia removal and glutamine synthesis. Furthermore Hep G2 cells could grow well in the absence of glutamine, which was necessarily required in mammalian cell cultures. These results prove that glutamine formation serves as the primary mechanism of detoxifying ammonia in hepatocyte cell lines as expected. In addition, it was demonstrated that ammonium level could be reduced 38% and that erythropoietin production increased 2-fold in the mixed culture of Hep G2 and recombinant CHO cells. PMID- 11027168 TI - Measurement and control of dissolved carbon dioxide in mammalian cell culture processes using an in situ fiber optic chemical sensor. AB - At high viable cell concentrations in large-scale mammalian cell culture processes, the accumulation of dissolved carbon dioxide (dCO(2), typically quantified as an equilibrium gas-phase concentration) becomes problematic as a result of low CO(2) removal rates at reduced surface-to-volume ratios. High dCO(2) concentrations have previously been shown to inhibit cell growth and product formation in mammalian cells and to alter the glycosylation pattern of recombinant proteins. Therefore, reliable monitoring and control of dCO(2) are important for successful large-scale operation. Off-line measurements by instruments such as blood gas analyzers (BGA) are constrained by the low frequency of data collection and cannot be used for on-line control. In a preliminary evaluation of the YSI 8500 in situ sensor, a response time (t(90%)) of 6 min, sensitivity of 0.5% CO(2) (3.6 mmHg), and linearity of measurement (R(2) = 0.9997) between the equivalent gas-phase partial pressure of 0-180 mmHg (0% and 25% CO(2)) were established. Measurements were found to be unaffected by culture pH and typical mammalian cell culture concentrations of glucose, glutamine, glutamate, lactate, and ammonium. The sensor withstood repeated sterilization and cleaning cycles. The reliability of this sensor was demonstrated in microcarrier-based Chinese hamster ovary (CHO) cell perfusion cultures at reactor scales of 30, 40, 340, and 2000 L and was successfully implemented in a dCO(2) control strategy using N(2) sparging. PMID- 11027169 TI - Osmoprotective effect of glycine betaine on thrombopoietin production in hyperosmotic Chinese hamster ovary cell culture: clonal variations. AB - When 23 recombinant Chinese hamster ovary (rCHO) cell clones were cultivated in hyperosmolar medium resulting from NaCl addition (533 mOsm/kg), their specific thrombopoietin (TPO) productivity (q(TPO)) was increased. However, due to depressed cell growth at elevated osmolality, no enhancement in the maximum TPO titer was made in batch cultures of all 23 clones. To test the feasibility of using glycine betaine, known as a strong osmoprotective compound, for improved TPO production in hyperosmotic rCHO cell cultures, hyperosmotic batch cultures of 23 clones were performed in the presence of 15 mM glycine betaine. Glycine betaine was found to have a strong osmoprotective effect on all 23 clones. Inclusion of 15 mM glycine betaine in hyperosmolar medium enabled 22 clones to grow at 542 mOsm/kg, where most clones could not grow in the absence of glycine betaine, but at a cost of reduced q(TPO). However, the relative decrease in q(TPO) varied significantly among clones. Thus, efficacy of the simultaneous use of hyperosmotic pressure and glycine betaine as a means to improve foreign protein production was variable among clones. Six out of 23 clones displayed more than a 40% increase in the maximum TPO titer in the hyperosmolar medium containing glycine betaine, compared with that in the standard medium with a physiological osmolality. Taken together, the results obtained here emphasize the importance of selection of clones for the successful use of hyperosmotic pressure and glycine betaine as an economical means to improve TPO production. PMID- 11027170 TI - Osmolarity effects on observed insect cell size after baculovirus infection are avoided using growth medium for sample dilution. AB - Rates of cell size increase are an important measure of success during the baculovirus infection process. Batch and fed batch cultures sustain large fluctuations in osmolarity that can affect the measured cell volume if this parameter is not considered during the sizing protocol. Where osmolarity differences between the sizing diluent and the culture broth exist, biased measurements of size are obtained as a result of the cell osmometer response. Spodoptera frugiperda (Sf9) cells are highly sensitive to volume change when subjected to a change in osmolarity. Use of the modified protocol with culture supernatants for sample dilution prior to sizing removed the observed error during measurement. PMID- 11027171 TI - Increased t-PA yields using ultrafiltration of an inhibitory product from CHO fed batch culture. AB - Fed-batch operation for the production of t-PA using Chinese Hamster Ovary (CHO) cells was optimized using serial and parallel experimentation. The feed, an isotonic concentrate, was improved to obtain 2- to 2.5-fold increases in integrated viable cell days versus batch. With a low glucose inoculum train, the viability index was further increased up to 4.5-fold. Hydrolysates were substituted for the amino acid portion of the concentrate with no significant change in fed-batch results. The concentrate addition rate was based on a constant 4 pmol/cell.day glucose uptake rate that maintained a relatively constant glucose concentration (approximately 3 mM). Increased viable cell indices did not lead to concomitant increases in t-PA concentrations compared to batch. The fed-batch concentrate and feeding strategy were shown to be effective in hybridoma culture, where a 4-fold increase in viable cell index yielded a 4 fold increase in antibody concentration. The half-life of t-PA decreased from 43 to 15 days with decreasing cell viability (from 92% to 71%), but this was not sufficient to explain the apparent t-PA threshold. Instead, the CHO results were explained by a reduction in t-PA production at higher extracellular t-PA concentrations that limited the fed-batch maximum at 35 mg/L for the cell line investigated. Analysis of both the total and t-PA mRNA levels revealed no response to increasing extracellular t-PA concentrations upon exogenous additions. Instead, intracellular t-PA levels were increased, revealing a possible secretory pathway limitation. A new reactor configuration was developed using an acoustic filter to retain the cells in the reactor while an ultrafiltration module stripped t-PA from the clarified medium before the permeate was returned to the reactor. By adding this harvesting step, the t-PA fed-batch production was increased over 2-fold, up to a yield of 80 mg/L. PMID- 11027172 TI - Effect of low inoculation density in the scale-up of insect cell cultures. AB - The scale-up of insect cell cultures and the production of baculovirus with these cultures is dependent on the inoculation density applied. The effect of applying a low inoculation on the specific growth rate and on the duration of the lag phase was tested. Three different cell lines, HzAm1, Ha2302, and Sf21 were tested in a total of five cell line/medium combinations. Growth in suspension culture was examined, and data obtained were fitted with the Gompertz equation. A significant decline in specific growth rate with decreasing inoculation density was observed in all cell line/medium combinations, except for HzAm1. No critical inoculation density, below which no growth would occur, was found. In suspension culture in shake flasks, an inoculation density of 5 x 10(4) cells/mL is achievable, without severely influencing the overall growth rate. A lower inoculation density in suspension culture results in less steps in the scale-up process and might be a tool in bypassing the viral passage effect. PMID- 11027173 TI - Using the Microcyte flow cytometer to monitor cell number, viability, and apoptosis in mammalian cell culture. AB - The Microcyte is a novel, portable flow cytometer based on diode laser technology whose use has been established for yeast and bacterial analysis. We present data that demonstrate its suitability for routine mammalian cell counting and viability determination. To extend its range of applications in the field of animal cell culture biotechnology, a test to determine the number of apoptotic cells present has been developed for use with the instrument. Apoptosis was induced in hybridoma cell cultures by treatment with camptothecin. Apoptotic cells were labeled with biotinylated Annexin V and then visualized using a streptavidin-allophycocyanin conjugate. Their numbers were counted, and the cell size of the apoptotic cell population was determined using the Microcyte. PMID- 11027174 TI - On-line monitoring of physiological parameters of insect cell cultures during the growth and infection process. AB - On-line monitoring of insect cell cultures used for the production of recombinant proteins with the baculovirus expression vector system (BEVS) provides valuable tools for the optimization, operation, and control of the production process. The relative permittivity (epsilon') and CO(2) evolution rates (CER) were measured on line using the biomass monitor and the infrared CO(2) analyzer, respectively. The growth and infection phases of two different cell lines, Spodoptera frugiperda (Sf-9) and Trichoplusia ni(High-5), were monitored using the above measurements. These in turn were correlated to the progress of the culture by using the off line measurements of protein produced, virus titer, and biovolume, which is the product of viable cell density and mean cell volume. The epsilon', CER, and the biovolume profiles were closely matched during the growth phase of cells when grown in a batch or fed batch culture. The relationship became more complex when the cultures were either in stationary phase or in the postinfection phase. The epsilon' profile was found to be a good indicator of the process of synchronous baculoviral infection, showing a plateau between 18 and 24 h postinfection (hpi), the period during which budded virus is produced, and a peak at approximately 48 hpi correlated to the onset of accelerated cell lysis. The CER profile continues to increase after the growth period with a peak around the 24 hpi period, after which there is a decline in the profile corresponding to release of virus as seen from virus titer determinations. This was examined for Sf-9 cultures under conditions of cell densities from 3 to 50 x 10(6) cells/mL and MOI values ranging from 0.001 to 1000. The profiles were found to be similar also in the case of the High-5 cells. Thus both measurements give reliable information regarding the physiological status of the cells as seen from their correlation to virus and protein production. A further combination of these with the off-line measured parameters such as the biovolume and metabolite concentrations can give a more detailed understanding of the process and help in the better design and automation of these processes. PMID- 11027175 TI - Biomass and aggregation analysis of human embryonic kidney 293 suspension cell cultures by particle size measurement. AB - A method has been developed to monitor the cell growth of aggregated human embryonic kidney 293 (HEK293) suspension cultures by measuring cumulative particle volume and the particle size distribution. This method employs a particle size analyzer that determines the size of individual particles by detecting their light obscuration (blockage) or scattering. Cell counts derived from the cumulative volume of the cell particle correlate well with manual cell counts from a hemacytometer at different stages of growth. This correlation was further confirmed by quantifying total cellular protein of the samples. Simultaneously, the aggregation state of the samples can also be monitored and mathematically described. Results from this study demonstrate that this simple and reproducible method allows the direct measurement of cumulative cell volume and the degree of cell aggregation, as well as an indirect assessment of cell counts. PMID- 11027176 TI - A comparison of different methods to determine the end of exponential growth in CHO cell cultures for optimization of scale-up. AB - Maximizing cell growth rate and cell yield are among the most important features of a successful mammalian cell culture production process. To minimize time and resources needed to scale up cell mass it is important to maintain the cultures in exponential growth at every scale. Here we report results comparing viable cell counts, packed cell volume, intracellular nucleotide ratios, cell cycle analysis, and on-line oxygen uptake rates (OUR) and optical density for the determination of the end of exponential growth to optimize transfer times during scale-up of CHO cell cultures. Viable cell concentration, packed cell volume, and relative abundance of cells in S-phase were not very reliable at determining the end of exponential growth during the process. In contrast, on-line determination of OUR and off-line determination of intracellular nucleotide ratios (U-ratio) were very sensitive to changes in growth rate, enabling clear determination of the end of exponential growth within a short time. Although on-line OUR was found to be the most convenient and fastest method, it is restricted to instrumented and continuously monitored cultures. In contrast the nucleotide method can be applied with any culture scale and condition but needs the availability of an operator running an HPLC system and takes about an hour from sampling to result. Optical density showed an inflection along with OUR and U-ratio but was less sensitive in determining the end of exponential growth. PMID- 11027177 TI - First steps in robot automation of sampling and sample management during cultivation of mammalian cells in pilot scale. AB - The robot automation of sampling and the subsequent treatment and storage of aliquots during mammalian cell cultivations was investigated. The complete setup, the development and testing of the sampling device, the robot arm, and the cell imaging system are described. The developed sampling device is directly coupled to a pilot bioreactor. It allows the computerized sterile filling of cell broth into 50 mL sample tubes. After each sampling the whole tubing system is steam sterilized. For further off-line treatment a robot takes the sample to the different devices. This robot is equipped with a camera and a force/torque sensor. A color-based object recognition guides the arm in a complex surrounding with different illumination situations, enabling the robot to load the sampling device with tubes and take the sample to further devices. For necessary pipetting and refilling we developed a computerized device. Cells are automatically stained and counted using an imaging system. The cell number and viability is automatically saved in a process control system together with the on-line parameters. During several cultivations in 20 and 100 L scale these main components of the automation strategy were successfully tested. PMID- 11027178 TI - A tool for modeling strategic decisions in cell culture manufacturing. AB - The development of a prototype tool for modeling manufacturing in a biopharmaceutical plant is discussed. A hierarchical approach to modeling a manufacturing process has been adopted to confer maximum user flexibility. The use of this framework for assessing the impact of manufacturing decisions on strategic technical and business indicators is demonstrated via a case study. In the case study, which takes the example of a mammalian cell culture process delivering a therapeutic for clinical trials, the dynamic modeling tool indicates how manufacturing options affect the demands on resources and the associated manufacturing costs. The example illustrates how the decision-support software can be used by biopharmaceutical companies to investigate the effects of working toward different strategic goals on the cost-effectiveness of the process, prior to committing to a particular option. PMID- 11027179 TI - Cell cycle progression in serum-free cultures of Sf9 insect cells: modulation by conditioned medium factors and implications for proliferation and productivity. AB - Cell cycle progression was studied in serum-free batch cultures of Spodoptera frugiperda (Sf9) insect cells, and the implications for proliferation and productivity were investigated. Cell cycle dynamics in KBM10 serum-free medium was characterized by an accumulation of 50-70% of the cells in the G(2)/M phase of the cell cycle during the first 24 h after inoculation. Following the cell cycle arrest, the cell population was redistributed into G(1) and in particular into the S phase. Maximum rate of proliferation (micro(N, max)) was reached 24-48 h after the release from cell cycle arrest, coinciding with a minimum distribution of cells in the G(2)/M phase. The following declining micro(N) could be explained by a slow increase in the G(2)/M cell population. However, at approximately 100 h, an abrupt increase in the amount of G(2)/M cells occurred. This switch occurred at about the same time point and cell density, irrespective of medium composition and maximum cell density. An octaploid population evolved from G(2)/M arrested cells, showing the occurrence of endoreplication in this cell line. In addition, conditioned medium factor(s) were found to increase micro(N,max), decrease the time to reach micro(N,max), and decrease the synchronization of cells in G(2)/M during the lag and growth phase. A conditioned medium factor appears to be a small peptide. On basis of these results we suggest that the observed cell cycle dynamics is the result of autoregulatory events occurring at key points during the course of a culture, and that entry into mitosis is the target for regulation. Infecting the Sf9 cells with recombinant baculovirus resulted in a linear increase in volumetric productivity of beta galactosidase up to 68-75 h of culture. Beyond this point almost no product was formed. Medium renewal at the time of infection could only partly restore the lost hypertrophy and product yield of cultures infected after the transition point. The critical time of infection correlated to the time when the mean population cell volume had attained a minimum, and this occurred 24 h before the switch into the G(2)/M phase. We suggest that the cell density dependent decrease in productivity ultimately depends on the autoregulatory events leading to G(2)/M cell cycle arrest. PMID- 11027180 TI - Cybernetic modeling and regulation of metabolic pathways in multiple steady states of hybridoma cells. AB - Hybridoma cells utilize a pair of complementary and partially substitutable substrates, glucose and glutamine, for growth. It has been shown that cellular metabolism shifts under different culture conditions. When those cultures at different metabolic states are switched to a continuous mode, they reach different steady states under the same operating conditions. A cybernetic model was constructed to describe the complementary and partial substitutable nature of substrate utilization. The model successfully predicted the metabolic shift and multiple steady-state behavior. The results are consistent with the experimental observation that the history of the culture affects the resulting steady state. PMID- 11027181 TI - Application of a serum-free medium for the growth of Vero cells and the production of reovirus. AB - Two strains of reovirus (serotype 1 Lang/TIL and serotype 3 Dearing/T3D) were propagated in Vero cells grown in stationary or agitated cultures in a serum-free medium, M-VSFM. Solid microcarriers (Cytodex-1) were used to support cell growth in agitated cultures with a normal doubling time of 25 h. Cell yields of 1 x 10(6) cells/mL were obtained from an inoculum of 2 x 10(5) cells/mL in 4 days in microcarrier cultures. The growth profile and cell yield was not significantly different from serum-supplemented cultures. The virus titer increased by 3-4 orders of magnitude over a culture period of 150 h. The maximum virus titer in stationary cultures reached >1 x 10(9) pfu/mL for both strains of reovirus in M VSFM. M-VSFM also supported high viral yields in microcarrier cultures. Both the specific productivity and final viral yield was higher in M-VSFM than serum supplemented cultures. The high viral productivity suggests that this is a suitable system for the production of reovirus as an oncolytic agent for human therapeutic use. PMID- 11027182 TI - Effects of culture parameters on the production of retroviral vectors by a human packaging cell line. AB - The use of retroviral vectors for human gene therapy requires the production of large quantities of high titer vector stocks. Maintaining high titers during the prolonged culture of packaging cells will require that critical parameters be controlled. The aim of this study was to determine which culture parameters critically affect the production/decay of retroviral vectors produced by the human packaging cell line FLYRD18/LNC-hB7. The stability of retroviral vectors released by this cell line was found to be temperature dependent (half-life of 6.9, 11.0, and 64.3 h when incubated at 37, 32, and 0 degrees C, respectively). Titers increased up to 10-fold when the packaging cells were cultured at 32 degrees C, compared to 37 degrees C, despite a decrease in cell yield (cell specific titers were 20-fold higher). Virus titers were also over 10-fold higher when the packaging cells were cultured in a reduced serum concentration (1%) compared to 5%. Retrovirus production at a range of pH levels revealed a significant decrease in virus titer at pH levels below 6.8 and above 7.2, optimum titers being achieved in cultures at pH 7.2. Dissolved oxygen levels in the range 20-80% did not significantly affect titers under the conditions tested. Finally, a packed bed system containing the packaging cells immobilized on porous microcarriers was shown to sustain the production of active retroviral vectors for over 1 month, in relatively large volumes. PMID- 11027183 TI - Monitoring adenovirus infections with on-line and off-line methods. AB - Several known process monitoring methods were tested for their efficacy in the detection of adenovirus infections. The methods that we explored include several indirect indications of viral infections, including metabolic rate analysis, secondary gauges of respiration, cell size measurement, cell number and cell viability determination, and changes in capacitance. Direct indications of the adenovirus infection were also applied, including total viral particle and infectious particle measurements, as well as a flow cytometry method for detecting infected cells. All of the methods tested in the study provide some positive indication of an adenovirus infection. Many of the methods require repeated sampling, which may limit their utility in a manufacturing process. All of the indirect measures of viral infection may be limited by the fact that they do not uniquely identify an infection. The simplest monitoring methods appear to be detection of changes in respiration or the capacitance of the culture, both of which seem to provide a clear indication of an infection. Further work will be required to demonstrate that these indications are characteristic of only a successful and productive adenovirus infection. PMID- 11027184 TI - 293SF metabolic flux analysis during cell growth and infection with an adenoviral vector. AB - Metabolic flux quantification of cell culture is becoming a crucial means to improve cell growth as well as protein and vector productions. The technique allows rapid determination of cell culture status, thus providing a tool for further feeding improvements. Herein, we report on key results of a metabolic investigation using 293 cells adapted to suspension and serum-free medium (293SF) during growth and infection with an adenoviral vector encoding the green fluorescence protein (GFP). The model developed contains 35 fluxes, which include the main fluxes of glycolysis, glutaminolysis, and amino acids pathways. It requires specific consumption and production rate measurements of amino acids, glucose, lactate, NH(3), and O(2), as well as DNA and total proteins biosynthesis rate measurements. Also, it was found that extracellular protein concentration measurement is important for flux calculation accuracy. With this model, we are able to describe the 293SF cell metabolism, grown under different culture conditions in a 3-L controlled bioreactor for batch and fed-batch with low glucose. The metabolism is also investigated during infection under two different feeding strategies: a fed-batch starting at the end of the growth phase and extending during infection without medium change and a fed-batch after infection following medium renewal. Differences in metabolism are observed between growth and infection, as well as between the different feeding strategies, thus providing a better understanding of the general metabolism. PMID- 11027185 TI - The lactate issue revisited: novel feeding protocols to examine inhibition of cell proliferation and glucose metabolism in hematopoietic cell cultures. AB - It is well established that cell proliferation in batch (unfed) hematopoietic cell cultures is greatly inhibited relative to that in cultures with feeding. What is not known, however, is the nature of this inhibition. On the basis of our observations in hematopoietic cultures that cell proliferation ceases when the lactate concentration ([lactate]) exceeds 20 mM (accompanied by a decrease in culture pH), we investigated the effect of lactate accumulation on cell proliferation, metabolism, and differentiation. We differ in our approach from previous efforts in that we have tried to more accurately recreate the manner in which lactate accumulates in culture by employing a daily feeding protocol in which [lactate] and/or pH in the fresh medium was adjusted to match the conditions prior to feeding. We conclude that the decrease in pH associated with lactate accumulation significantly inhibits both cell proliferation and metabolism. Although inhibition in cultures with high [lactate] and low pH is similar to that in unfed cultures, pH control in unfed cultures does not alleviate the inhibition, indicating that other inhibitory factors are also present. Thus, pH control is necessary, but not sufficient, to eliminate inhibition of cell growth and metabolism in unfed hematopoietic cell cultures. We also conclude that high [lactate] and low pH have little effect on cell differentiation in fed cultures, although there is evidence to suggest that low pH may play a role in monocyte differentiation in unfed cultures. PMID- 11027186 TI - Comparison of chondrogensis in static and perfused bioreactor culture. AB - As a result of the low yield of cartilage from primary patient harvests and a high demand for autologous cartilage for reconstructive surgery and structural repair, primary explant cartilage must be augmented by tissue engineering techniques. In this study, chondrocytes seeded on PLLA/PGA scaffolds in static culture and a direct perfusion bioreactor were biochemically and histologically analyzed to determine the effects of fluid flow and media pH on matrix assembly. A gradual media pH change was maintained in the bioreactor within 7.4-6.96 over 2 weeks compared to a more rapid decrease from 7.4 to 6.58 in static culture over 3 days. Seeded scaffolds subjected to 1 microm/s flow demonstrated a 118% increase (p < 0.05) in DNA content, a 184% increase (p < 0.05) in GAG content, and a 155% (p < 0.05) increase in hydroxyproline content compared to static culture. Distinct differences were noted in tissue morphology, including more intense staining for proteoglycans by safranin-O and alignment of cells in the direction of media flow. Culture of chondrocyte seeded matrices thus offers the possibility of rapid in vitro expansion of donor cartilage for the repair of structural defects, tracheal injury, and vascularized tissue damage. PMID- 11027187 TI - Biocompatibility of HEMA copolymers designed for treatment of CNS diseases with polymer-encapsulated cells. AB - Surrounding the cells with a semipermeable polymeric membrane allows transplanting unmatched xenogeneic cells without a risk of their rejection. We prepared and tested several 2-hydroxyethyl methacrylate (HEMA) copolymers with alkyl methacrylates or acrylates to find out which was the most valuable for cell encapsulation. On the basis of optimum physical properties and good results of cytotoxicity tests, HEMA-EMA copolymer was chosen as a suitable candidate for encapsulation and immunoprotection of xenogeneic cells before their grafting into the central nervous system (CNS). To characterize the biocompatibility of p(HEMA co-EMA) copolymer in the CNS, we implanted microcapsules made of this hydrogel into the brains of adult rats that were allowed to survive for 0.5, 1, 3, 6, and 9 months. Analysis of histological sections containing the implantation site was aimed at assessment of the cellular density at the implant-brain interface and identification of cell types participating in a tissue reaction. Our results indicated that the tissue reaction that was observed was caused largely by the implantation procedure because HLA-DR- and GSI-B4-positive macrophages/microglia infiltrated mainly the implantation channel. The number of these cells declined with time, which was true also for GFAP-positive reactive astrocytes, as well as for foreign body giant cells. The amount of connective tissue components surrounding the implanted microcapsules increased only slightly. These findings indicated that p(HEMA-co-EMA) hydrogel was well tolerated after implantation in the brain. PMID- 11027188 TI - Computational model for effects of ligand/receptor binding properties on interleukin-2 trafficking dynamics and T cell proliferation response. AB - Multisubunit cytokine receptors such as the heterotrimeric receptor for interleukin-2 (IL-2) are ubiquitous in hematopoeitic cell types of importance in biotechnology and are crucial regulators of cell proliferation and differentiation behavior. Dynamics of cytokine/receptor endocytic trafficking can significantly impact cell responses through effects of receptor down-regulation and ligand depletion, and in turn are governed by ligand/receptor binding properties. We describe here a computational model for trafficking dynamics of the IL-2 receptor (IL-2R) system, which is able to predict T cell proliferation responses to IL-2. This model comprises kinetic equations describing binding, internalization, and postendocytic sorting of IL-2 and IL-2R, including an experimentally derived dependence of cell proliferation rate on these properties. Computational results from this model predict that IL-2 depletion can be reduced by decreasing its binding affinity for the IL-2R betagamma subunit relative to the alpha subunit at endosomal pH, as a result of enhanced ligand sorting to recycling vis-a-vis degradation, and that an IL-2 analogue with such altered binding properties should exhibit increased potency for stimulating the T cell proliferation response. These results are in agreement with our recent experimental findings for the IL-2 analogue termed 2D1 [Fallon, E. M. et al. J. Biol. Chem. 2000, 275, 6790-6797]. Thus, this type of model may enable prediction of beneficial cytokine/receptor binding properties to aid development of molecular design criteria for improvements in applications such as in vivo cytokine therapies and in vitro hematopoietic cell bioreactors. PMID- 11027190 TI - The JECH gallery: a call for public health photographs PMID- 11027189 TI - Development and qualification of a novel virus removal filter for cell culture applications PMID- 11027191 TI - Good intentions and received wisdom are not enough. PMID- 11027192 TI - Researching the changing social pattern of cardiovascular diseases. PMID- 11027194 TI - The association between violence victimisation and common symptoms in Swedish women. AB - STUDY OBJECTIVE: To investigate the association between violence and abuse suffered by women during childhood or adult life, and the manifestation of a high level of common physical and mental symptoms. DESIGN, SETTING AND PARTICIPANTS: A questionnaire was sent to a random population of women, 40 to 50 years of age, living in a rural Swedish community. The response rate was 81.7 per cent (397 women). Odds ratios were used to estimate bivariate associations between the experience of violence/abuse and common symptoms. Multiple logistic regression analyses were used to test for confounding and effect modification. MAIN RESULTS: The experience of violence or abuse during childhood was reported by 32.2 per cent of the women, while 15.6 per cent reported being abused as an adult. In both cases, these experiences reached statistical significance in their association with a high level of common symptoms (OR=1.67; 95% CI 1. 08, 2.49 and OR=2.26; 95%CI 1.30, 3.92, respectively). The associations between childhood and as well adult experience of violence or abuse and common symptoms were largely independent of potential confounders such as unemployment, job strain, social support, and sense of coherence. The combined exposure to adult violence/abuse and low psychosocial coping resources, such as low social support or a low level of sense of coherence, considerably increased the odds ratio for common symptoms and a synergistic effect seemed to exist. CONCLUSION: Violence or abuse experience is an important factor when considering illness manifestations in terms of common symptoms in women 40 to 50 years of age. PMID- 11027193 TI - Social distribution of cardiovascular disease risk factors: change among men in England 1984-1993. AB - OBJECTIVE: To investigate change in the social distribution of some of the main risk factors for cardiovascular disease in men in England during a period when inequality in cardiovascular disease mortality widened DESIGN: Age standardised comparison of the social distribution of seven known risk factors for cardiovascular disease (body mass index, waist to hip ratio, systolic and diastolic blood pressure, consumption of fresh green vegetables, leisure time exercise, cigarette smoking and levels of social support) in two large cross sectional representative samples of the English population. SUBJECTS: Men aged 20 64 years in the 1984 Health and Lifestyle Survey (excluding Scotland and Wales) first sweep and the 1993 Health Survey for England. MAIN OUTCOME MEASURES: Mean values of continuous variables; age adjusted proportions of categorical variables; change in the relative index of inequality for each risk factor. RESULTS: The overall prevalence of cardiovascular disease risk factors improved during the period in which cardiovascular disease mortality was falling. The social distribution of cardiovascular disease risk factors, in contrast, did not become more extreme. Increases in the relative index of inequality for angina from 1.75 to 1.86, for eating vegetables less than once a day from 1.76 in 1984 to 1.96 in 1993, and an apparently larger increase in inequality of social support, from 1.92 to 2.53 were not statistically significant. In most cases the degree of inequality in risk factors tended to narrow non-significantly: for example the relative index of inequality fell from 5.02 in 1984 to 3.07 in 1993 for systolic blood pressure, from 5.60 to 4.29 for current smoking and from 6.24 to 4.19 for eating other than wholemeal bread as the main form of bread in the diet. The two statistically significant changes in inequality were in the direction of narrowing inequality: from a relative index of inequality of 2.12 to 0.90 for diastolic blood pressure (p<0.01) and from 19.3 to 0.87 (p<0.01) for psychological distress as indicated by the General Health Questionnaire. CONCLUSIONS: Healthier lifestyle options have not been adopted at a significantly faster rate by middle class than working class people over this time period. At the population level the change in risk factors is consistent with falling cardiovascular mortality. The change in the social distribution of risk factors within the population, however, shows little or no relation to the pattern of widening inequality in cardiovascular mortality. This may be because the effect is lagged, or because the adoption of healthier behaviour confers greater benefits on those in higher socioeconomic status groups. PMID- 11027195 TI - Childhood leukaemia in areas with different radon levels: a spatial and temporal analysis using GIS. AB - OBJECTIVES: To evaluate the relation between exposure to ground radon levels and leukaemia among children using existing population and disease registers. DESIGN: Ecological correlation study. SETTING: The county of Ostergotland in Sweden. METHODS: Every child born in the county between 1979 and 1992 was mapped to the property centroid coordinates by linking addresses in the population and property registers. Population maps were overlaid with radon maps and exposure at birth and each subsequent year was quantified as high, normal, low or unknown. This was analysed with data from the tumour registry. Standardised mortality ratios (SMRs) were calculated using the age and sex specific rates for Sweden for the year 1995. RESULTS: 90 malignancies occurred among 53 146 children (498 887 person years) who formed the study population. SMRs for acute lymphatic leukaemia (ALL) among children born in high, normal and low risk areas were 1.43, 1.17 and 0.25 respectively. The relative risk for the normal risk group and high risk group as compared with the low risk group was 4.64 (95% CI 1.29, 28.26) and 5. 67 (95% CI 1.06, 42.27). The association between ALL and continued residence at normal or high risk areas showed a similar trend. No association between radon risk levels and any other malignancy was seen. CONCLUSION: Children born in and staying at areas where the risk from ground radon has been classified as low are less likely to develop ALL than those born in areas classified as normal and high risk. PMID- 11027196 TI - Fatigue in the Danish general population. Influence of sociodemographic factors and disease. AB - OBJECTIVE: To measure the levels of fatigue in the general population, and to examine how disease and sociodemographic factors influence fatigue. DESIGN: Cross sectional questionnaire study in the Danish general population. SUBJECTS: A random, age stratified sample of 1608 people aged 20-77 with an equal gender distribution (response rate 67%). MAIN OUTCOME MEASURES: Five fatigue scales from the questionnaire Multidimensional Fatigue Inventory: General Fatigue, Physical Fatigue, Reduced Activity, Reduced Motivation and Mental Fatigue. RESULTS: Fatigue scores were skewed towards absence of fatigue. The General Fatigue and Physical Fatigue scales showed the highest fatigue levels while the Reduced Motivation scale showed lowest levels. Gender differences in fatigue scores were small, but the variability among women was higher-that is, more women had high scores. A multiple linear regression analysis showed that respondents of low social status and respondents with a depression had high fatigue scores on all scales, independent of other factors. Chronic somatic disease had an independent direct effect on Mental Fatigue, but for the rest of the scales, the effect of somatic disease depended on age, gender and/or whether the person was living alone. For example, General and Physical Fatigue decreased with age among healthy people, whereas scores on these scales increased with age among those with a somatic disease. CONCLUSIONS: Physical and mental diseases play essential parts for the level of fatigue and as modulators of the associations between sociodemographic factors and fatigue. These interactions should be taken into account in future research on fatigue and sociodemographic factors and when data from clinical studies are compared with normative data from the general population. PMID- 11027197 TI - Occupational risk factors for ill health in Gulf veterans of the United Kingdom. AB - OBJECTIVES: To study the association between occupational factors specific to the Armed Forces (rank, functional roles, Service, regular or reservist status and deployment factors) and symptomatic health problems in Gulf veterans, after sociodemographic and lifestyle factors have been accounted for. DESIGN: A postal cross sectional survey of randomly selected UK Gulf veterans was conducted six to seven years after the Gulf conflict. Physical ill health was measured using the Fatigue Questionnaire and a measure of the Centers for Disease Control and Prevention (CDC) multi-symptom syndrome. Psychological ill health was measured using the General Health Questionnaire and a post-traumatic stress measure. SETTING: Population of servicemen who were serving in the UK Armed Forces during the Gulf conflict between 1 September 1990 and 30 June 1991. PARTICIPANTS: 3297 Gulf veterans. MAIN RESULTS: In multivariate logistic regression, there was an inverse relation between higher rank and psychological and physical ill health (test of trend: General Health Questionnaire, p=0.004; post-traumatic stress, p=0.002; fatigue, p=0.015; CDC case, p=0.002). Having left the Armed Forces was associated with a two to three times increase in reporting ill health. Of the deployment factors, there was a weak association between being deployed as an individual reinforcement in a combat role and post-traumatic stress but there was no association between receiving pre-deployment training or post-deployment leave and ill health. Marital status and smoking were associated with psychological and physical ill health. CONCLUSIONS: Rank was the main occupational factor associated with both psychological and physical ill health in Gulf veterans. This may parallel the associations between socioeconomic status and morbidity in civilian populations. Ill health seems to be greater in those who return to civilian life. Sociodemographic factors also seem to be important in ill health in Gulf veterans. PMID- 11027198 TI - Excess type 2 diabetes in African-American women and men aged 40-74 and socioeconomic status: evidence from the Third National Health and Nutrition Examination Survey. AB - OBJECTIVE: To examine whether socioeconomic status (SES) explains differences in the prevalence of type 2 diabetes between African-American and non-Hispanic white women and men. DESIGN: Cross sectional study of diabetes prevalence, SES, and other risk factors ascertained by physical examination and interview. SETTING: Interviews were conducted in subjects' homes; physical examinations were conducted in mobile examination centres. PARTICIPANTS: 961 African-American women, 1641 non-Hispanic white women, 839 African-American men and 1537 non Hispanic white men, aged 40 to 74 years, examined in the Third National Health and Nutrition Examination Survey (NHANES III), a representative sample of the non institutionalised civilian population of the United States, 1988-1994. MAIN RESULTS: Among women, African-American race/ethnicity was associated with an age adjusted odds ratio of 1.76 (95% confidence intervals 1.21, 2.57), which was reduced to 1.42 (95% confidence intervals 0.95, 2.13) when poverty income ratio was controlled. Controlling for education or occupational status had minimal effects on this association. When other risk factors were controlled, race/ethnicity was not significantly associated with type 2 diabetes prevalence. Among men, the age adjusted odds ratio associated with African-American race/ethnicity was 1.43 (95% confidence intervals 1.03, 1.99). Controlling for SES variables only modestly affected the odds ratio for African/American race/ethnicity among men, while adjusting for other risk factors increased the racial/ethnic differences. CONCLUSIONS: Economic disadvantage may explain much of the excess prevalence of type 2 diabetes among African-American women, but not among men. PMID- 11027199 TI - Images and reflections: photography in the pursuit of public health. PMID- 11027201 TI - How accurately do adult sons and daughters report and perceive parental deaths from coronary disease ? AB - OBJECTIVES: To describe how adult sons and daughters report and perceive parental deaths from heart disease DESIGN: Two generation family study. SETTING: West of Scotland. SUBJECTS: 1040 sons and 1298 daughters aged 30-59 from 1477 families, whose fathers and mothers were aged 45-64 in 1972-76 and have been followed up for mortality over 20 years. OUTCOME: Perception of a "family weakness" attributable to heart disease. RESULTS: 26% of sons and daughters had a parent who had died of coronary heart disease (CHD). The proportion was higher in older offspring (+18% per 10 year age difference) and in manual compared with non manual groups (+37%). Eighty nine per cent of parental deaths from CHD were correctly reported by offspring. Only 23% of sons and 34% of daughters with at least one parent who had died of CHD considered that they had a family weakness attributable to heart disease. Perceptions of a family weakness were higher when one or both parents had died of CHD, when parental deaths occurred at a younger age, in daughters compared with sons and in offspring in non-manual compared with manual occupations. CONCLUSIONS: Only a minority of sons and daughters with experience of a parent having died from CHD perceive this in terms of a family weakness attributable to heart disease. Although men in manual occupations are most likely to develop CHD, they are least likely to interpret a parental death from CHD in terms of a family weakness. Health professionals giving advice to patients on their familial risks need to be aware of the difference between clinical definitions and lay perceptions of a family history of heart disease. PMID- 11027200 TI - Breast cancer hypothesis: a single cause for the majority of cases. AB - STUDY OBJECTIVE: The main cause of breast cancer remains unknown. Numerous causal factors or predisposing conditions have been proposed, but account for only a small percentage of the total disease. The current search for multiple causes is unavailing. This report explores whether any single aetiological agent may be responsible for the majority of cases, and attempts to define its properties. METHODS: Examination of all relevant epidemiological and biological evidence. MAIN RESULTS: Genetic inheritance is not the main cause of breast cancer because most cases are sporadic, there is a low prevalence of family history, and genetically similar women have differing rates after migration. Environmental exposure, such as pollution by industrialisation, is not a major cause, as deduced from a spectrum of epidemiological data. The possibility of infection as cause is not persuasive as there is no direct biological evidence and no epidemiological support. Oestrogen status is closely related to breast cancer risk, but there are numerous inconsistencies and paradoxes. It is suggested that oestrogens are not the proximate agent but are promoters acting in concert with the causal agent. Dietary factors, and especially fat, are associated with the aetiology of breast cancer as shown by intervention and ecological correlation studies, but the evidence from case-control and cohort studies is inconsistent and contradictory. CONCLUSIONS: The hypothesis that best fits the epidemiological data is that dietary fat is not itself the causal agent, but produces depletion of an essential factor that is normally protective against the development of breast cancer. Many of the observed inconsistencies in the epidemiology are explainable if deficiency of this agent is permissive for breast cancer to develop. Some properties of the putative agent are outlined, and research investigations proposed. PMID- 11027203 TI - Community exposures to chemical incidents: development and evaluation of the first environmental public health surveillance system in europe. AB - OBJECTIVE: To describe the frequency, nature and location of acute chemical incidents in Wales, and the morbidity in employees, emergency responders and the general public who were exposed. DESIGN: Active multi-agency community-based surveillance system. SETTING: Wales, 1993-5. MAIN OUTCOME MEASURES: Frequency, nature and location of incidents, populations potentially exposed and with symptoms. RESULTS: Most of the 402 incidents identified were not associated with sites governed by the Control of Industrial Major Accident Hazard Regulations but with smaller industrial sites and commercial premises. About two in every thousand of the estimated 236 000 members of the public considered to be at risk from exposure reported symptoms, which were mainly nausea, headaches, and irritation of the eye, skin and respiratory tract. The most commonly reported chemicals that members of the public were exposed to were smoke toxins, miscellaneous organics, toxic gases and flammable gases. A health authority was reported to be involved in only 34 (8%) of the incidents and in only 3 of the 29 incidents where more than 100 members of the public were exposed. CONCLUSION: A geographically defined, multi-agency surveillance system can identify high risk locations and types of incidents, together with the chemicals most likely to be involved. Such ongoing surveillance information is essential for appropriate policy making, emergency planning, operational management and training. PMID- 11027202 TI - The health of the Roma people: a review of the published literature. AB - BACKGROUND: The Roma people originated in northern India and have been known in Europe for nearly a thousand years. For much of that time they have been the subjects of discrimination and oppression, culminating in the extermination of half a million Roma in the Nazi death camps. While it is widely believed that the health of Roma people is often poorer than the majority population, these inequalities remain largely unresearched. METHODS: Published literature on the health of the Roma people was identified using Medline. Opinion pieces were excluded, as were papers relating to anthropometry and to genetic markers. The resultant papers were analysed by country of study and by disease type or care group. RESULTS: Some 70% of papers identified related to just three countries; Spain and the Czech and Slovak Republics. Much literature concentrates upon communicable disease or reproductive health. The limited evidence suggests increased morbidity from non-communicable disease, but there is little published on this topic. Evidence on health care, though fragmentary, suggests poorer access to health services and uptake of preventative care. DISCUSSION: Published research on the health needs of the Roma population is sparse. The topics that have received attention suggest a focus on concepts of contagion or social Darwinism, indicating a greater concern with the health needs of the majority populations with which they live. There is a need for both further research into the health of Roma people; with particular emphasis on non-communicable disease; and also for interventions that improve Roma health. Such research must, however, be handled with sensitivity, recognising the social and political context of the society concerned. PMID- 11027204 TI - The relation between cholesterol and haemorrhagic or ischaemic stroke in the Renfrew/Paisley study. PMID- 11027206 TI - Dilemmas in community mental health practice: choice or control PMID- 11027205 TI - Cardiovascular reactions to psychological stress: the influence of demographic variables. PMID- 11027207 TI - Deficiency of complement defense protein CD59 may contribute to neurodegeneration in Alzheimer's disease. AB - Complement defense 59 (CD59) is a cell surface glycophosphoinositol (GPI) anchored protein that prevents complement membrane attack complex (MAC) assembly. Here, we present evidence from ELISA assays that CD59 protein levels are significantly decreased in the frontal cortex and hippocampus of Alzheimer's disease (AD) compared with nondemented elderly (ND) patients, whereas complement component 9, a final component to form MAC, is significantly increased. To further confirm the CD59 deficit, PI-specific phospholipase C (PIPLC) was used to cleave the CD59 GPI anchor at the cell surface in intact slices from AD and ND cortex. CD59 released by PIPLC cleavage was significantly reduced in AD compared with ND samples. By the use of a ribonuclease protection technique, amyloid beta peptide was found to downregulate CD59 expression at the mRNA level, suggesting a partial explanation of CD59 deficits in the AD brain. To evaluate the pathophysiological significance of CD59 alterations in neurons, we exposed cultured NT2 cells, which normally underexpress CD59, and NT2 cells transfected to overexpress CD59 to homologous human serum. Lactic acid dehydrogenase assays revealed significant complement-induced cell lysis in CD59-underexpressing NT2 cells and significant protection from such lysis in CD59-overexpressing NT2 cells. Moreover, cells expressing normal levels of CD59 showed no evidence of MAC assembly or damage after exposure to homologous serum, whereas pretreatment of these cells with a CD59-neutralizing antibody resulted in MAC assembly at the cell surface and morphological damage. Taken together, these data suggest that CD59 deficits may play a role in the neuritic losses characteristic of AD. PMID- 11027208 TI - Mechanism of interleukin-1- and tumor necrosis factor alpha-dependent regulation of the alpha 1-antichymotrypsin gene in human astrocytes. AB - The expression of alpha(1)-antichymotrypsin (ACT) is significantly enhanced in affected brain regions in Alzheimer's disease. This serine proteinase inhibitor specifically colocalizes with filamentous beta-amyloid deposits and recently has been shown to influence both formation and destabilization of beta-amyloid fibrils. In the brain, ACT is expressed in astrocytes, and interleukin-1 (IL-1), tumor necrosis factor alpha (TNF), oncostatin M (OSM), and IL-6/soluble IL-6 receptor complexes control synthesis of this inhibitor. Here, we characterize a molecular mechanism responsible for both IL-1 and TNF-induced expression of ACT gene in astrocytes. We identify the 5' distal IL-1/TNF-responsive enhancer of the ACT gene located 13 kb upstream of the transcription start site. This 413-bp-long enhancer contains three elements, two of which bind nuclear factor kB (NF-kB) and one that binds activating protein 1 (AP-1). All of these elements contribute to the full responsiveness of the ACT gene to both cytokines, as determined by deletion and mutational analysis. The 5' NF-kB high-affinity binding site and AP 1 element contribute most to the enhancement of gene transcription in response to TNF and IL-1. In addition, we demonstrate that the 5' untranslated region of the ACT mRNA does not contribute to cytokine-mediated activation. Finally, we find that overexpression of the NF-kB inhibitor (IkB) totally inhibits any activation mediated by the newly identified IL-1/TNF enhancer of the ACT gene. PMID- 11027209 TI - Mutants of a temperature-sensitive two-P domain potassium channel. AB - Within the Caenorhabditis elegans genome there exist at least 42 genes encoding TWK (two-P domain K(+)) channels, potassium channel subunits that contain two pore regions and four transmembrane domains. We now report the first functional characterization of a TWK channel from C. elegans. Although potassium channels have been reported to be activated by a variety of factors, TWK-18 currents increase dramatically with increases in temperature. Two mutant alleles of the twk-18 gene confer uncoordinated movement and paralysis in C. elegans. Expression of wild-type and mutant TWK-18 channels in Xenopus oocytes showed that mutant channels express much larger potassium currents than wild-type channels. Promoter green fluorescent protein fusion experiments indicate that TWK-18 is expressed in body wall muscle. Our genetic and physiological data suggest that the movement defects observed in mutant twk-18 animals may be explained by an increased activity of the mutant TWK-18 channels. PMID- 11027210 TI - Light and glutamate-induced degradation of the circadian oscillating protein BMAL1 during the mammalian clock resetting. AB - Recently discovered mammalian clock genes are believed to compose the core oscillator, which generates the circadian rhythm. BMAL1/CLOCK heterodimer is the essential positive element that drives clock-related transcription and self sustaining oscillation by a negative feedback mechanism. We examined BMAL1 protein expression in the rat suprachiasmatic nuclei (SCN) by immunoblot analysis. Anti-BMAL1 antiserum raised against rBMAL1 recognized 70 kDa mBMAL1b and detected a similar immunoreactivity (IR) as a major band in rat brains. Robust circadian BMAL1-IR oscillations with nocturnal peaks were detected in the SCN during a light/dark cycle and under constant darkness. A short duration light exposure at night acutely reduced BMAL1-IR in the SCN during photoentrainment. This might be attributable to the degradation of BMAL1 protein. Application of glutamate and NMDA to the SCN slices at projected night, a procedure mimicking photic phase delay shift, also acutely reduced BMAL1-IR in a similar manner. A rapid decrease of BMAL1 protein suggests that BMAL1 protein might be implicated in the light-transducing pathway within the SCN. PMID- 11027211 TI - Abnormal GABAA receptor-mediated currents in dorsal root ganglion neurons isolated from Na-K-2Cl cotransporter null mice. AB - We have recently disrupted Slc12a2, the gene encoding the secretory Na-K-2Cl cotransporter in mice (NKCC1) (Delpire et al., 1999). Gramicidin perforated-patch and whole-cell recordings were performed to study GABA-induced currents in dorsal root ganglion (DRG) neurons isolated from wild-type and homozygote NKCC1 knock out mice. In wild-type DRG neurons, strong GABA-evoked inward current was observed at the resting membrane potential, suggesting active accumulation of Cl( ) in these cells. This GABA-induced current was blocked by picrotoxin, a GABA(A) receptor blocker. The strong Cl(-) accumulation that gives rise to depolarizing GABA responses is caused by Na-K-2Cl cotransport because reduction of external Cl(-) or application of bumetanide induced a decrease in [Cl(-)](i), whereas an increase in external K(+) caused an apparent [Cl(-)](i) accumulation. In contrast to control neurons, little or no net current was observed at the resting membrane potential in homozygote NKCC1 mutant DRG neurons. E(GABA) was significantly more negative, demonstrating the absence of Cl(-) accumulation in these cells. Application of bumetanide induced a positive shift of E(GABA), suggesting the presence of an outward Cl(-) transport mechanism. In agreement with an absence of GABA depolarization in DRG neurons, behavioral analysis revealed significant alterations in locomotion and pain perception in the knock-out mouse. Our results clearly demonstrate that the Na-K-2Cl cotransporter is responsible for [Cl(-)](i) accumulation in DRG neurons and that via regulation of intracellular Cl(-), the Na-K-2Cl cotransporter participates in the modulation of GABA neurotransmission and sensory perception. PMID- 11027212 TI - Activity-dependent maintenance of long-term potentiation at visual cortical inhibitory synapses. AB - Neural activity producing a transient increase in intracellular Ca(2+) concentration can induce long-term potentiation (LTP) at visual cortical inhibitory synapses similar to those seen at various excitatory synapses. Here we report that low-frequency neural activity is required to maintain LTP at these inhibitory synapses. Inhibitory responses of layer 5 cells evoked by layer 4 stimulation were studied in developing rat visual cortical slices under a pharmacological blockade of excitatory synaptic transmission using intracellular and whole-cell recording methods. Although LTP induced by high-frequency stimulation (HFS) persisted while test stimulation was applied at 0.1 Hz, it was not maintained in approximately two-thirds of cells after test stimulation was stopped for 30 min. In the rest of the cells, LTP seemed to be maintained by spontaneous presynaptic spikes, because presynaptic inhibitory cells discharged spontaneously in our experimental condition and because LTP was totally abolished by a temporary application of Na(+) channel blockers. Experiments applying various Ca(2+) channel blockers and Ca(2+) chelators after HFS demonstrated that LTP maintenance was mediated by presynaptic Ca(2+) entries through multiple types of high-threshold Ca(2+) channels, which activated Ca(2+)-dependent reactions different from those triggering transmitter release. The Ca(2+) entries associated with action potentials seemed to be regulated by presynaptic K(+) channels, presumably large-conductance Ca(2+)-activated K(+) channels, because the application of blockers for these channels facilitated LTP maintenance. In addition, noradrenaline facilitated the maintenance of LTP. These findings demonstrate a new mechanism by which neural activity regulates the continuation and termination of LTP at visual cortical inhibitory synapses. PMID- 11027213 TI - Two novel doubletime mutants alter circadian properties and eliminate the delay between RNA and protein in Drosophila. AB - Phosphorylation is an important feature of pacemaker organization in Drosophila. Genetic and biochemical evidence suggests involvement of the casein kinase I homolog doubletime (dbt) in the Drosophila circadian pacemaker. We have characterized two novel dbt mutants. Both cause a lengthening of behavioral period and profoundly alter period (per) and timeless (tim) transcript and protein profiles. The PER profile shows a major difference from the wild-type program only during the morning hours, consistent with a prominent role for DBT during the PER monomer degradation phase. The transcript profiles are delayed, but there is little effect on the protein accumulation profiles, resulting in the elimination of the characteristic lag between the mRNA and protein profiles. These results and others indicate that light and post-transcriptional regulation play major roles in defining the temporal properties of the protein curves and suggest that this lag is unnecessary for the feedback regulation of per and tim protein on per and tim transcription. PMID- 11027214 TI - NF-kappa B signaling promotes both cell survival and neurite process formation in nerve growth factor-stimulated PC12 cells. AB - Nerve growth factor binds to the TrkA and p75(NTR) (p75) and generates signals leading to neuronal cell survival, differentiation, and programmed cell death. Here we describe a series of experiments involving selective activation of either TrkA or p75 in which distinct cell-signaling intermediates promote different cellular consequences. We analyzed pheochromocytoma 12 (PC12) cells stably expressing chimeras consisting of the extracellular domain of PDGF receptor (PDGFR) fused to the transmembrane and cytoplasmic segments of p75 or TrkA. Because PC12 cells lack endogenous PDGFR, addition of PDGF to these cell lines permits selective activation of the p75 or TrkA responses without stimulating endogenous receptors. Although both p75 and TrkA activated nuclear factor-kappaB (NF-kappaB), we show that distinct proximal-signaling intermediates are used by each receptor. A dominant-negative mutant of TRAF6 blocked p75- but not TrkA mediated induction of NF-kappaB. Conversely a dominant-negative mutant of Shc inhibited TrkA but not p75 activation of NF-kappaB. Both of these distinct signaling pathways subsequently converge, leading to activation of the IkappaB kinase complex. Moreover, the activation of NF-kappaB by these distinct pathways after stimulation of either TrkA or p75 leads to different physiological consequences. Blocking p75-mediated activation of NF-kappaB by ecdysone-inducible expression of a nondegradable mutant of IkappaBalpha significantly enhanced apoptosis. In contrast, blocking NF-kappaB induction via TrkA significantly inhibited neurite process formation in PC12 cells. Together these findings indicate that, although both of these receptors lead to the activation of NF kappaB, they proceed via distinct proximal-signaling intermediates and contribute to different cellular outcomes. PMID- 11027215 TI - C-Terminal alternative splicing changes the gating properties of a human spinal cord calcium channel alpha 1A subunit. AB - The calcium channel alpha(1A) subunit gene codes for proteins with diverse structure and function. This diversity may be important for fine tuning neurotransmitter release at central and peripheral synapses. The alpha(1A) C terminus, which serves a critical role in processing information from intracellular signaling molecules, is capable of undergoing extensive alternative splicing. The purpose of this study was to determine the extent to which C terminal alternative splicing affects some of the fundamental biophysical properties of alpha(1A) subunits. Specifically, the biophysical properties of two alternatively spliced alpha(1A) subunits were compared. One variant was identical to an isoform identified previously in human brain, and the other was a novel isoform isolated from human spinal cord. The variants differed by two amino acids (NP) in the extracellular linker between transmembrane segments IVS3 and IVS4 and in two C-terminal regions encoded by exons 37 and 44. Expression in Xenopus oocytes demonstrated that the two variants were similar with respect to current voltage relationships and the voltage dependence of steady-state activation and inactivation. However, the rates of activation, inactivation, deactivation, and recovery from inactivation were all significantly slower for the spinal cord variant. A chimeric strategy demonstrated that the inclusion of the sequence encoded by exon 44 specifically affects the rate of inactivation. These findings demonstrate that C-terminal structural changes alone can influence the way in which alpha(1A) subunits respond to a depolarizing stimulus and add to the developing picture of the C terminus as a critical domain in the regulation of Ca(2+) channel function. PMID- 11027217 TI - The excitatory neuronal network of rat layer 4 barrel cortex. AB - Sensory whiskers are mapped to rodent layer 4 somatosensory cortex as discrete units termed barrels, which can be visualized at high resolution in living brain slices. Both anatomical and physiological properties of the layer 4 neuronal network can thus be investigated in the context of the functional boundaries of this sensory map. Large-scale confinement of neuronal arbors to single barrels was suggested by restricted lateral diffusion of DiI across septa between barrels. Morphological analysis of dendritic and axonal arborizations of individual excitatory neurons showed that neuronal processes remain within the barrel of origin through polarization toward the center of the barrel. Functionally, the large-scale properties of the neuronal network were investigated through mapping the spatial extent of field EPSPs, which were found to attenuate at barrel borders. This ensemble property of a layer 4 barrel was further investigated by analyzing the connectivity of pairs of excitatory neurons with respect to the locations of the somata. Approximately one-third of the excitatory neurons within the same barrel were synaptically coupled. At the septum between adjacent barrels the connectivity dropped rapidly, and very few connections were found between neurons located in adjacent barrels. Each layer 4 barrel is thus composed of an excitatory neuronal network, which to a first order approximation, acts independently of its neighbors. PMID- 11027216 TI - Cocaine and antidepressant-sensitive biogenic amine transporters exist in regulated complexes with protein phosphatase 2A. AB - Presynaptic transporter proteins regulate the clearance of extracellular biogenic amines after release and are important targets for multiple psychoactive agents, including amphetamines, cocaine, and antidepressant drugs. Recent studies reveal that dopamine (DA), norepinephrine (NE), and serotonin (5-HT) transporters (DAT, NET, and SERT, respectively) are rapidly regulated by direct or receptor-mediated activation of cellular kinases, particularly protein kinase C (PKC). With SERTs, PKC activation results in activity-dependent transporter phosphorylation and sequestration. Protein phosphatase 1/2A (PP1/PP2A) inhibitors, such as okadaic acid (OA) and calyculin A, also promote SERT phosphorylation and functional downregulation. How kinase, phosphatase, and transporter activities are linked mechanistically is unclear. In the present study, we found that okadaic acid sensitive phosphatase activity is enriched in SERT immunoprecipitates from human SERT stably transfected cells. Moreover, blots of these immunoprecipitates reveal the presence of PP2A catalytic subunit (PP2Ac), findings replicated using brain preparations. Whole-cell treatments with okadaic acid or calyculin A diminished SERT/PP2Ac associations. Phorbol esters, which trigger SERT phosphorylation, also diminish SERT/PP2Ac associations, effects that can be blocked by PKC antagonists as well as the SERT substrate 5-HT. Similar transporter/PP2Ac complexes were also observed in coimmunoprecipitation studies with NETs and DATs. Our findings provide evidence for the existence of regulated heteromeric assemblies involving biogenic amine transporters and PP2A and suggest that the dynamic stability of these complexes may govern transporter phosphorylation and sequestration. PMID- 11027218 TI - CD45 opposes beta-amyloid peptide-induced microglial activation via inhibition of p44/42 mitogen-activated protein kinase. AB - Reactive microglia have been suggested to play a role in the Alzheimer's disease (AD) process, and previous studies have shown that expression of CD45, a membrane bound protein-tyrosine phosphatase (PTP), is elevated in microglia in AD brain compared with controls. To investigate the possible role of CD45 in microglial responsiveness to beta-amyloid (Abeta) peptides, we first co-treated primary cultured microglia with a tyrosine phosphatase inhibitor [potassium bisperoxo (1,10-phenanthroline) oxovanadate (phen), 5 micrometer] and freshly solubilized Abeta peptides (1000 nm). Data show synergistic induction of microglial activation as evidenced by tumor necrosis factor alpha (TNF-alpha) production and nitric oxide (NO) release, both of which we show to be dependent on activation of p44/42 mitogen-activated protein kinase (MAPK). Furthermore, co-treatment with phen and Abeta peptides results in microglia-induced neuronal cell injury. Stimulation of microglial CD45 by anti-CD45 antibody markedly inhibits these effects via inhibition of p44/42 MAPK, suggesting that CD45 is a negative regulator of microglial activation. Accordingly, primary cultured microglia from CD45-deficient mice demonstrate hyper-responsiveness to Abeta, as evidenced by TNF-alpha release, NO production, and neuronal injury after stimulation with Abeta peptides. As a validation of these findings in vivo, brains from a transgenic mouse model of AD [transgenic Swedish APP-overexpressing (Tg APP(sw)) mice] deficient for CD45 demonstrate markedly increased production of TNF-alpha compared with Tg APP(sw) mice. Taken together, these results suggest that therapeutic agents that stimulate the CD45 PTP signaling pathway may be effective in suppressing microglial activation associated with AD. PMID- 11027219 TI - Differential localization of divalent metal transporter 1 with and without iron response element in rat PC12 and sympathetic neuronal cells. AB - Two isoforms of divalent metal transporter 1 (DMT1) (Nramp2 and DCT1) are encoded by two mRNA species, one of which contains an iron response element (IRE) motif in the 3'-noncoding region. The subcellular distribution of the two isoforms of DMT1 is distinct, and the -IRE species accumulates in the nucleus of neuronal or neuronal-like cells. Reverse transcription-PCR and Western blot analysis of PC12 cells reveals that these cells express both forms of DMT1. Immunofluorescence and immunoblotting studies, using immunospecific antibodies to the -IRE form of DMT1, demonstrate that this form of the transporter, in PC12 cells, is predominantly localized in the nucleus, cell membrane, and neurites with only weak staining of the cell body. Studies using antibodies to the +IRE form indicate that this species of DMT1 is distributed within vesicles in the cell body and neurite projections, with minimal nuclear staining. Similar staining patterns are observed for the two forms of DMT1 in cultures of sympathetic ganglion neurons isolated from perinatal rat pups. To determine whether nuclear localization of the -IRE form of DMT1 is constrained to neuronal or neuronal-like cells, immunocytochemical studies were performed with human embryonic kidney 293T (HEK293T), HEP2G hepatoma and medulloblastoma, and rat Schwann cells. The -IRE specific antibodies stained nuclei from medulloblastoma, whereas little nuclear staining was observed with HEK293T, hepatoma, or Schwann cells. The unexpected finding that the -IRE species of DMT1 selectively accumulates in the nucleus of neuronal and neuronal-like cells leads us to postulate that the two proteins may have different functions in vivo. PMID- 11027220 TI - Dual roles for c-Jun N-terminal kinase in developmental and stress responses in cerebellar granule neurons. AB - c-Jun N-terminal kinases (JNKs) typically respond strongly to stress, are implicated in brain development, and are believed to mediate neuronal apoptosis. Surprisingly, however, JNK does not respond characteristically to stress in cultured cerebellar granule (CBG) neurons, a widely exploited CNS model for studies of death and development, despite the regulation of its substrate c-Jun. To understand this anomaly, we characterized JNK regulation in CBG neurons. We find that the specific activity of CBG JNK is elevated considerably above that from neuron-like cell lines (SH-SY5Y, PC12); however, similar elevated activities are found in brain extracts. This activity does not result from cellular stress because the stress-activated protein kinase p38 is not activated. We identify a minor stress-sensitive pool of JNK that translocates with mitogen-activated protein kinase kinase-4 (MKK4) into the nucleus. However, the major pool of total activity is cytoplasmic, residing largely in the neurites, suggesting a non nuclear role for JNK in neurons. A third JNK pool is colocalized with MKK7 in the nucleus, and specific activities of both increase during neuritogenesis, nuclear JNK activity increasing 10-fold, whereas c-Jun expression and activity decrease. A role for JNK during differentiation is supported by modulation of neuritic architecture after expression of dominant inhibitory regulators of the JNK pathway. Channeling of JNK signaling away from c-Jun during differentiation is consistent with the presence in the nucleus of the JNK/MKK7 scaffold protein JNK interacting protein, which inhibits JNK-c-Jun interaction. We propose a model in which distinct pools of JNK serve different functions, providing a basis for understanding multifunctional JNK signaling in differentiating neurons. PMID- 11027221 TI - G-Protein types involved in calcium channel inhibition at a presynaptic nerve terminal. AB - The inhibition of presynaptic calcium channels via G-protein-dependent second messenger pathways is a key mechanism of transmitter release modulation. We used the calyx-type nerve terminal of the chick ciliary ganglion to examine which G proteins are involved in the voltage-sensitive inhibition of presynaptic N-type calcium channels. Adenosine caused a prominent inhibition of the calcium current that was totally blocked by pretreatment with pertussis toxin (PTX), consistent with an exclusive involvement of G(o)/G(i) in the G-protein pathway. Immunocytochemistry was used to localize these G-protein types to the nerve terminal and its transmitter release face. We used two approaches to test for modulation by other G-protein types. First, we treated the terminals with ligands for a variety of G-protein-linked neurotransmitter receptor types that have been associated with different G-protein families. Although small inhibitory effects were observed, these could all be eliminated by PTX, indicating that in this terminal the G(i) family is the sole transmitter-induced G-protein inhibitory pathway. Second, we examined the kinetics of calcium channel inhibition by uncaging the nonselective and irreversible G-protein activator GTPgammaS, bypassing the receptors. A large fraction of the rapid GTPgammaS-induced inhibition persisted, consistent with a G(o)/G(i)-independent pathway. Immunocytochemistry identified G(q), G(11), G(12), and G(13) as potential PTX insensitive second messengers at this terminal. Thus, our results suggest that whereas neurotransmitter-mediated calcium channel inhibition is mainly, and possibly exclusively, via G(o)/G(i), other rapid PTX-insensitive G-protein pathways exist that may involve novel, and perhaps transmitter-independent, activating mechanisms. PMID- 11027222 TI - Akt-mediated survival of oligodendrocytes induced by neuregulins. AB - Neuregulins have been implicated in a number of events in cells in the oligodendrocyte lineage, including enhanced survival, mitosis, migration, and differentiation. At least two signaling pathways have been shown to be involved in neuregulin signaling: the phosphatidylinositol (PI)-3 kinase and the mitogen activated protein kinase pathways. In the present studies, we examined the signaling pathway involved in the survival function of heregulin, focusing on heregulin-induced changes in Akt activity in cultured glial cells, and the consequences of Akt activation in cells in the oligodendrocyte lineage. Heregulin binds erbB receptors, and in our studies, primary cultures of both oligodendrocyte progenitor cells and differentiating oligodendrocytes expressed erbB2, erbB3, and erbB4 receptors. In C6 glioma cells and primary cultures of oligodendrocytes, heregulin induced time- and dose-dependent Akt phosphorylation at Ser(473) in a wortmannin-sensitive manner. To investigate further the signaling pathway for heregulin in glial cells, BAD was overexpressed in C6 glioma cells. In these cells, heregulin induced phosphorylation of BAD at Ser(136). Apoptosis of oligodendrocyte progenitor cells induced by growth factor deprivation was effectively blocked by heregulin in a wortmannin-sensitive manner. Overexpression of dominant negative Akt but not of wild-type Akt by adenoviral gene transfer in primary cultures of both oligodendrocytes and their progenitors induced significant apoptosis through activation of the caspase cascade. The present data suggest that the survival function of heregulin is mediated through the PI-3 kinase/Akt pathway in cells in the oligodendrocyte lineage and that the Akt pathway may be quite important for survival of cells in this lineage. PMID- 11027223 TI - Impairment of long-term potentiation and associative memory in mice that overexpress extracellular superoxide dismutase. AB - Reactive oxygen species, including superoxide, generally are considered neurotoxic molecules whose effects can be alleviated by antioxidants. Different from this view, we show that scavenging of superoxide with an antioxidant enzyme is associated with deficits in hippocampal long-term potentiation (LTP), a putative neural substrate of memory, and hippocampal-mediated memory function. Using transgenic mice that overexpress extracellular superoxide dismutase (EC SOD), a superoxide scavenger, we found that LTP was impaired in hippocampal area CA1 despite normal LTP in area CA3. The LTP impairment in area CA1 could be reversed by inhibition of EC-SOD. In addition, we found that EC-SOD transgenic mice exhibited impaired long-term memory of fear conditioning to contextual cues despite exhibiting normal short-term memory of the conditioning experience. These findings strongly suggest that superoxide, rather than being considered exclusively a neurotoxic molecule, should also be considered a signaling molecule necessary for normal neuronal function. PMID- 11027224 TI - Enhanced spinal nociceptin receptor expression develops morphine tolerance and dependence. AB - The tolerance and dependence after chronic medication with morphine are thought to be representative models for studying the plasticity, including the remodeling of neuronal networks. To test the hypothesis that changes in neuronal plasticity observed in opioid tolerance or dependence are derived from increased activity of the anti-opioid nociceptin system, the effects of chronic treatments with morphine were examined using nociceptin receptor knock-out (NOR(-/-)) mice and a novel nonpeptidic NOR antagonist, J-113397, which shows a specific and potent NOR antagonist activity in in vitro [(35)S]GTPgammaS binding assay and in vivo peripheral nociception test. The NOR(-/-) mice showed marked resistance to morphine analgesic tolerance without affecting morphine analgesic potency in tail pinch and tail-flick tests. The NOR(-/-) mice also showed marked attenuation of morphine-induced physical dependence, manifested as naloxone-precipitated withdrawal symptoms after repeated morphine treatments. Similar marked attenuation of morphine tolerance was also observed by single subcutaneous (10 mg/kg) or intrathecal (1 nmol) injection of J-113397, which had been given 60 min before the test in morphine-treated ddY mice. However, the intracerebroventricular injection (up to 3 nmol) did not affect the tolerance. On the other hand, morphine dependence was markedly attenuated by J-113397 that had been subcutaneously given 60 min before naloxone challenge. There was also observed a parallel enhancement of NOR gene expression only in the spinal cord during chronic morphine treatments. Together, these findings suggest that the spinal NOR system develops anti-opioid plasticity observed on morphine tolerance and dependence. PMID- 11027225 TI - Evidence that helix-loop-helix proteins collaborate with retinoblastoma tumor suppressor protein to regulate cortical neurogenesis. AB - The retinoblastoma tumor suppressor protein (pRb) family is essential for cortical progenitors to exit the cell cycle and survive. In this report, we test the hypothesis that pRb collaborates with basic helix-loop-helix (bHLH) transcription factors to regulate cortical neurogenesis, taking advantage of the naturally occurring dominant-inhibitory HLH protein Id2. Overexpression of Id2 in cortical progenitors completely inhibited the induction of neuron-specific genes and led to apoptosis, presumably as a consequence of conflicting differentiation signals. Both of these phenotypes were rescued by coexpression of a constitutively activated pRb mutant. In contrast, Id2 overexpression in postmitotic cortical neurons affected neither neuronal gene expression nor survival. Thus, pRb collaborates with HLHs to ensure the coordinate induction of terminal mitosis and neuronal gene expression as cortical progenitors become neurons. PMID- 11027226 TI - Evidence that different cation chloride cotransporters in retinal neurons allow opposite responses to GABA. AB - GABA gating an anion channel primarily permeable to chloride can hyperpolarize or depolarize, depending on whether the chloride equilibrium potential (E(Cl)) is negative or positive, respectively, to the resting membrane potential (E(rest)). If the transmembrane Cl(-) gradient is set by active transport, those neurons or neuronal regions that exhibit opposite responses to GABA should express different chloride transporters. To test this, we immunostained retina for the K-Cl cotransporter (KCC2) that normally extrudes chloride and for the Na-K-Cl cotransporter (NKCC) that normally accumulates chloride. KCC2 was expressed wherever E(Cl) is either known or predicted to be negative to E(rest) (ganglion cells, bipolar axons, and OFF bipolar dendrites), whereas NKCC was expressed wherever E(Cl) is either known or predicted to be positive to E(rest) (horizontal cells and ON bipolar dendrites). Thus, in the retina, the opposite effects of GABA on different cell types and on different cellular regions are probably primarily determined by the differential targeting of these two chloride transporters. PMID- 11027227 TI - Facial visceral motor neurons display specific rhombomere origin and axon pathfinding behavior in the chick. AB - In the chick embryo, facial motor neurons comprise branchiomotor and visceral motor subpopulations, which innervate branchial muscles and parasympathetic ganglia, respectively. Although facial motor neurons are known to develop within hindbrain rhombomere 4 (r4) and r5, the precise origins of branchiomotor and visceral motor neuron subpopulations are unclear. We investigated the organization and axon pathfinding of these motor neurons using axonal tracing and rhombomere transplantation in quail-chick chimeras. Our results show that a large majority of branchiomotor neurons originate in r4 but that a cohort of these neurons undergoes a caudal migration from r4 into r5. By contrast, visceral motor neurons develop exclusively in r5. We found that a striking property of facial visceral motor neurons is the ability of their axons to navigate back to appropriate ganglionic targets in the periphery after heterotopic transplantation. These results complement previous studies in which heterotopic facial branchiomotor neurons sent axons to their correct, branchial arch, target. By contrast, when trigeminal branchiomotor neurons were transplanted heterotopically, we found that they were unable to pathfind correctly, and instead projected to an inappropriate target region. Thus, facial and trigeminal motor neuron populations have different axon pathfinding characteristics. PMID- 11027228 TI - Mice lacking specific nicotinic acetylcholine receptor subunits exhibit dramatically altered spontaneous activity patterns and reveal a limited role for retinal waves in forming ON and OFF circuits in the inner retina. AB - Before phototransduction, spontaneous activity in the developing mammalian retina is required for the appropriate patterning of retinothalamic connections, and there is growing evidence that this activity influences the development of circuits within the retina itself. We demonstrate here that the neural substrate that generates waves in the mouse retina develops through three distinct stages. First, between embryonic day 16 and birth [postnatal day 0 (P0)], we observed both large, propagating waves inhibited by nicotinic acetylcholine receptor (nAChR) antagonists and small clusters of cells displaying nonpropagating, correlated calcium increases that were independent of nAChR activation. Second, between P0 and P11, we observed only larger propagating waves that were abolished by toxins specific to alpha3 and beta2 subunit-containing nAChRs. Third, between P11 and P14 (eye opening) we observed propagating activity that was abolished by ionotropic glutamate receptor antagonists. The time course of this developmental shift was dramatically altered in retinas from mice lacking the beta2 nAChR subunit or the beta2 and beta4 subunits. These retinas exhibited a novel circuit at P0, no spontaneous correlated activity between P1 and P8, and the premature induction at P8 of an ionotropic glutamate receptor-based circuit. Retinas from postnatal mice lacking the alpha3 nAChR subunit exhibited spontaneous, correlated activity patterns that were similar to those observed in embryonic wild-type mice. In alpha3-/- and beta2-/- mice, the development and distribution of cholinergic neurons and processes and the density of retinal ganglion cells (RGCs) and the gross segregation of their dendrites into ON and OFF sublaminae were normal. However, the refinement of individual RGC dendrites is delayed. These results indicate that retinal waves mediated by nAChRs are involved in, but not required for, the development of neural circuits that define the ON and OFF sublamina of the inner plexiform layer. PMID- 11027229 TI - Differential expression of COUP-TFI, CHL1, and two novel genes in developing neocortex identified by differential display PCR. AB - Genes that control the specification and differentiation of the functionally specialized areas of the mammalian neocortex are likely expressed across the developing neocortex in graded or restricted patterns. To search for such genes we have performed a PCR-based differential display screen using RNAs from rostral neocortex, which included the primary motor area, and caudal neocortex, which included the primary visual area, of embryonic day 16 rats. We identified 82 differentially expressed gene fragments. Secondary screening by in situ hybridization confirmed that five fragments, representing four genes, are differentially expressed across developing rat neocortex. Two of the genes, chick ovalbumin upstream transcription factor I (COUP-TFI) and close homolog of L1 (CHL1), have been cloned previously, but their differential expression in cortex has not been reported. Sequences from the other two fragments suggest that they represent novel genes. The expression patterns include graded, restricted, and discontinuous expression with abrupt borders that might correlate with those of areas. The differential expression patterns of all four genes are established before the arrival of thalamocortical afferents, suggesting that they are independent of thalamic influence, and could direct or reflect arealization. In addition, COUP-TFI and CHL1 exhibit dynamic expression patterns that undergo substantial changes after thalamocortical afferents invade the cortical plate, suggesting that thalamic axons may influence their later expression. Postnatally, COUP-TFI is most prominently expressed in layer 4, in both rats and mice, and CHL1 is expressed in layer 5. COUP-TFI expression in cortex, and in ventral telencephalon and dorsal thalamus, suggests several possible causes for the loss of layer 4 neurons and the reduced thalamocortical projection reported in COUP TFI knock-out mice. PMID- 11027230 TI - Semaphorin 3A is required for guidance of olfactory axons in mice. AB - Semaphorin 3A (Sema3A) is a membrane-associated secreted protein that has chemorepulsive properties for neuropilin-1 (npn-1)- expressing axons. Although mice lacking the Sema3A protein display skeletal abnormalities and heart defects, most axonal projections in the CNS develop normally. We show here that Sema3A is expressed in the lamina propria surrounding the olfactory epithelium (OE) and by ensheathing cells in the nerve layer of the ventral olfactory bulb (OB) throughout development. Subsets of sensory neurons expressing npn-1 are distributed throughout the OE and extend fibers to the developing OB. In wild type mice, npn-1-positive (npn-1(+)) axons extend to lateral targets in the rostral OB and medial targets in the caudal OB, avoiding regions expressing Sema3A. In Sema3A homozygous mutant mice, many npn-1(+) axons are misrouted into and through the ventral nerve layer, beginning as early as embryonic day 13 and continuing at least until birth. At postnatal day 0, npn-1(+) glomeruli are atypically located in the ventral OB of Sema3A(-/-) mice, indicating that aberrant axon trajectories are not corrected during development and that connections are made in inappropriate target regions. In addition, subsets of OCAM(+) axons that normally project to the ventrolateral OB and some lactosamine containing glycan(+) axons that normally target the ventral OB are also misrouted in Sema3A mutants. These observations indicate that Sema3A expression by ensheathing cells plays an important role in guiding olfactory axons into specific compartments of the OB. PMID- 11027231 TI - A transgenic mouse model for inducible and reversible dysmyelination. AB - Oligodendrocytes are glial cells devoted to the production of myelin sheaths. Myelination of the CNS occurs essentially after birth. To delineate both the times of oligodendrocyte proliferation and myelination, as well as to study the consequence of dysmyelination in vivo, a model of inducible dysmyelination was developed. To achieve oligodendrocyte ablation, transgenic animals were generated that express the herpes virus 1 thymidine kinase (HSV1-TK) gene under the control of the myelin basic protein (MBP) gene promoter. The expression of the MBP-TK transgene in oligodendrocytes is not toxic on its own; however, toxicity can be selectively induced by the systemic injection of animals with nucleoside analogs, such as FIAU [1-(2-deoxy-2-fluoro-beta-delta-arabinofuranosyl)-5-iodouracil]. This system allows us to control the precise duration of the toxic insult and the degree of ablation of oligodendrocytes in vivo. We show that chronic treatment of MBP-TK mice with FIAU during the first 3 postnatal weeks triggers almost a total depletion of oligodendrocytes in the CNS. These effects are accompanied by a behavioral phenotype characterized by tremors, seizures, retarded growth, and premature animal death. We identify the period of highest oligodendrocytes division in the first 9 postnatal days. Delaying the beginning of FIAU treatments results in different degrees of dysmyelination. Dysmyelination in MBP-TK mice is always accompanied by astrocytosis. Thus, this transgenic line provides a model to study the events occurring during dysmyelination of various intensities. It also represents an invaluable tool to investigate remyelination in vivo. PMID- 11027232 TI - p75 is important for axon growth and schwann cell migration during development. AB - Mice lacking the low-affinity neurotrophin receptor p75 have multiple peripheral neural deficits. Here we examined the developmental nature of these deficiencies. Peripheral axons in p75 -/- embryos were severely stunted and poorly arborized from embryonic day 11.5 (E11.5) to E14.5. In vitro, neurite outgrowth from the dorsal root ganglia was significantly decreased in the p75 -/- embryos at E12.5, suggesting that stunted axonal growth in the embryo may result in part from defects in neurite elongation. Additionally, Schwann cell marker S100beta immunoreactivity was decreased or absent along the growing axons of the ophthalmic branch from the trigeminal ganglia in p75 -/- embryos. Electron microscopy studies of the axons of the trigeminal ganglion at E13.5 revealed that in the p75 mutant embryo, nerve bundles were highly impaired and that coverage of the growing axons by Schwann cell cytoplasm was substantially reduced. In vitro, Schwann cell migration from the dorsal root ganglia was significantly decreased in the p75 -/- embryos at E12.5, suggesting that the lack of S100beta staining and Schwann cell coverage in the p75 mutant results from a deficit in Schwann cell migration. These results provide evidence that p75 is important in the developing embryo for regulating axon growth and arborization and for Schwann cell migration. PMID- 11027233 TI - Serotonin depletion in vivo inhibits the branching of olfactory projection neurons in the lobster deutocerebrum. AB - Serotonin depletion during embryogenesis has been shown previously to retard the growth of the olfactory and accessory lobes of the lobster deutocerebrum (Benton et al., 1997). The present study was undertaken to determine whether morphological changes in the interneurons innervating these lobes contribute to this growth retardation. We examined the effects of in vivo serotonin depletion using 5,7-dihydroxytryptamine (5,7-DHT) on the morphology of the olfactory projection neurons, one of two major classes of interneurons that innervate both lobes. Intracellular dye fills of olfactory projection neurons in normal embryos showed that each neuron extensively innervates either the olfactory or accessory lobe before projecting to neuropil regions in the protocerebrum. In embryos injected with 5,7-DHT, however, the deutocerebral arbors of 13.5% of the olfactory projection neurons examined were either markedly reduced compared with normal neurons or absent. Affected neurons also exhibited a number of additional aberrant morphological features suggesting that these neurons represent cells that were affected during their initial morphogenesis. Olfactory projection neurons with aberrant morphologies were also encountered, although less frequently (7.5% of the neurons examined), in control (sham-injected) embryos indicating that the sham injections can affect the development of the brain. This observation provides insights into the nature of effects seen in control embryos in previous experiments (Benton et al., 1997). The results of the present study indicate that in vivo serotonin depletion inhibits the branching of olfactory projection neurons and suggest, therefore, that one of the functions of serotonin during normal development is to promote the ingrowth of these neurons into the deutocerebral neuropils. PMID- 11027234 TI - Estrous changes in responses of rat gracile nucleus neurons to stimulation of skin and pelvic viscera. AB - Multi- and single-unit recording was performed in the gracile nucleus in urethane anesthetized rats to examine estrous variations in responses of its neurons to brushing the hindquarters and mechanical stimulation of the uterus, vaginal canal, cervix, and colon. Six rats each were studied in each of the four estrous stages: proestrus (P), estrus (E), metestrus (M), and diestrus (D). The magnitude of multi-unit responses to gentle brushing of the perineum, hip, and tail, but not the foot and leg, was significantly greater during proestrus than during other stages. Of 70 single units responsive to brush, 56 (80%) responded to stimulation of at least one viscus. Although this percentage did not change with estrous stage, the direction and latency of some responses did. Pressure on the cervix evoked significantly more inhibitory (vs excitatory) responses in P than in E and M, and the response latency was significantly longer in D and P than in E and M. The direction of response to vaginal distention did not change with estrous stage, but response latency was significantly longer in D than in P and E. Uterine distention evoked significantly more inhibitory responses in D than in P, with no estrous changes in latency. Responses to colon distention did not change. These variations in both magnitude of response to tactile stimulation and characteristics of response to stimulation of reproductive organs, but not the colon, correlate with changes in mating behaviors of the female rat, suggesting that the gracile nucleus is a component of neural systems that control reproductive behaviors. PMID- 11027235 TI - Corticotropin-releasing factor increases in vitro firing rates of serotonergic neurons in the rat dorsal raphe nucleus: evidence for activation of a topographically organized mesolimbocortical serotonergic system. AB - In vivo studies suggest that the stress-related neuropeptide corticotropin releasing factor (CRF) modulates serotonergic neurotransmission. To investigate the underlying mechanisms for this interaction, the present study examined the effects of CRF in vitro on dorsal raphe neurons that displayed electrophysiological and pharmacological properties consistent with a serotonergic phenotype. In the presence of either 1 or 2 mm Ca(2+), perfusion of ovine CRF or rat/human CRF rapidly and reversibly increased firing rates of a subpopulation (19 of 70, 27%) of serotonergic neurons predominantly located in the ventral portion of the dorsal raphe nucleus. For a given responsive neuron, the excitatory effects of CRF were reproducible, and there was no tachyphylaxis. Excitatory effects were dose-dependent (over the range of 0.1-1.6 micrometer) and were completely absent after exposure to the competitive CRF receptor antagonists alpha-helical CRF(9-41) or rat/human [d-Phe(12), Nle(21, 38), alpha-Me-Leu(37)] CRF(12-41). Both the proportion of responsive neurons and the magnitude of excitatory responses to CRF in the ventral portion of the caudal dorsal raphe nucleus were markedly potentiated in slices prepared from animals previously exposed to isolation and daily restraint stress for 5 d. Immunohistochemical staining of the recorded slices revealed close associations between CRF immunoreactive varicose axons and tryptophan hydroxylase-immunoreactive neurons in the area of the recordings, providing anatomical evidence for potential direct actions of CRF on serotonergic neurons. The electrophysiological properties and the distribution of responsive neurons within the dorsal raphe nucleus are consistent with the hypothesis that endogenous CRF activates a topographically organized mesolimbocortical serotonergic system. PMID- 11027236 TI - Coincident activation of NMDA and dopamine D1 receptors within the nucleus accumbens core is required for appetitive instrumental learning. AB - The nucleus accumbens, a brain structure ideally situated to act as an interface between corticolimbic information-processing regions and motor output systems, is well known to subserve behaviors governed by natural reinforcers. In the accumbens core, glutamatergic input from its corticolimbic afferents and dopaminergic input from the ventral tegmental area converge onto common dendrites of the medium spiny neurons that populate the accumbens. We have previously found that blockade of NMDA receptors in the core with the antagonist 2-amino-5 phosphonopentanoic acid (AP-5; 5 nmol) abolishes acquisition but not performance of an appetitive instrumental learning task (Kelley et al., 1997). Because it is currently hypothesized that concurrent dopamine D(1) and glutamate receptor activation is required for long-term changes associated with plasticity, we wished to examine whether the dopamine system in the accumbens core modulates learning via NMDA receptors. Co-infusion of low doses of the D(1) receptor antagonist SCH-23390 (0.3 nmol) and AP-5 (0.5 nmol) into the accumbens core strongly impaired acquisition of instrumental learning (lever pressing for food), whereas when infused separately, these low doses had no effect. Infusion of the combined low doses had no effect on indices of feeding and motor activity, suggesting a specific effect on learning. We hypothesize that co-activation of NMDA and D(1) receptors in the nucleus accumbens core is a key process for acquisition of appetitive instrumental learning. Such an interaction is likely to promote intracellular events and gene regulation necessary for synaptic plasticity and is supported by a number of cellular models. PMID- 11027238 TI - Emotional responses to pleasant and unpleasant olfactory, visual, and auditory stimuli: a positron emission tomography study. AB - Neural correlates of responses to emotionally valenced olfactory, visual, and auditory stimuli were examined using positron emission tomography. Twelve volunteers were scanned using the water bolus method. For each sensory modality, regional cerebral blood flow (rCBF) during presentation of both pleasant and unpleasant stimuli was compared with that measured during presentation of neutral stimuli. During the emotionally valenced conditions, subjects performed forced choice pleasant and unpleasant judgments. During the neutral conditions, subjects were asked to select at random one of a two key-press buttons. All stimulations were synchronized with inspiration, using an airflow olfactometer, to present the same number of stimuli for each sensory modality. A no-stimulation control condition was also performed in which no stimulus was presented. For all three sensory modalities, emotionally valenced stimuli led to increased rCBF in the orbitofrontal cortex, the temporal pole, and the superior frontal gyrus, in the left hemisphere. Emotionally valenced olfactory and visual but not auditory stimuli produced additional rCBF increases in the hypothalamus and the subcallosal gyrus. Only emotionally valenced olfactory stimuli induced bilateral rCBF increases in the amygdala. These findings suggest that pleasant and unpleasant emotional judgments recruit the same core network in the left hemisphere, regardless of the sensory modality. This core network is activated in addition to a number of circuits that are specific to individual sensory modalities. Finally, the data suggest a superior potency of emotionally valenced olfactory over visual and auditory stimuli in activating the amygdala. PMID- 11027237 TI - Nav2/NaG channel is involved in control of salt-intake behavior in the CNS. AB - Na(v)2/NaG is a putative sodium channel, whose physiological role has long been an enigma. We generated Na(v)2 gene-deficient mice by inserting the lacZ gene. Analysis of the targeted mice allowed us to identify Na(v)2-producing cells by examining the lacZ expression. Besides in the lung, heart, dorsal root ganglia, and Schwann cells in the peripheral nervous system, Na(v)2 was expressed in neurons and ependymal cells in restricted areas of the CNS, particularly in the circumventricular organs, which are involved in body-fluid homeostasis. Under water-depleted conditions, c-fos expression was markedly elevated in neurons in the subfornical organ and organum vasculosum laminae terminalis compared with wild-type animals, suggesting a hyperactive state in the Na(v)2-null mice. Moreover, the null mutants showed abnormal intakes of hypertonic saline under both water- and salt-depleted conditions. These findings suggest that the Na(v)2 channel plays an important role in the central sensing of body-fluid sodium level and regulation of salt intake behavior. PMID- 11027239 TI - Hypocretin-1 modulates rapid eye movement sleep through activation of locus coeruleus neurons. AB - The hypocretins (hcrts), also known as orexins, are two recently identified excitatory neuropeptides that in rat are produced by approximately 1200 neurons whose cell bodies are located in the lateral hypothalamus. The hypocretins/orexins have been implicated in the regulation of rapid eye movement (REM) sleep and the pathophysiology of narcolepsy. In the present study, we investigated whether the locus coeruleus (LC), a structure receiving dense hcrtergic innervation, which is quiescent during REM sleep, might be a target for hcrt to regulate REM sleep. Local administration of hcrt1 but not hcrt2 in the LC suppressed REM sleep in a dose-dependent manner and increased wakefulness at the expense of deep, slow-wave sleep. These effects were blocked with an antibody that neutralizes hcrt binding to hcrt receptor 1. In situ hybridization and immunocytochemistry showed the presence of hcrt receptor 1 but not the presence of hcrt receptor 2 in the LC. Iontophoretic application of hcrt1 enhanced the firing rate of LC neurons in vivo, and local injection of hcrt1 into the LC induced the expression of c-fos in the LC area. We propose that hcrt receptor 1 in the LC is a key target for REM sleep regulation and might be involved in the pathophysiological mechanisms of narcolepsy. PMID- 11027240 TI - High-frequency synchronization of neuronal activity in the subthalamic nucleus of parkinsonian patients with limb tremor. AB - It has been hypothesized that in Parkinson's disease (PD) there is increased synchronization of neuronal firing in the basal ganglia. This study examines the discharge activity of 121 pairs of subthalamic nucleus (STN) neurons in nine PD patients undergoing functional stereotactic mapping. Four patients had a previous pallidotomy. A double microelectrode setup was used to simultaneously record from two neurons separated by distances as small as 250 micrometer. In the six patients who had limb tremor during the recording session (n = 76 pairs), the discharge pattern of 12 pairs of tremor cells (TCs) was found to be coherent at the frequency of the limb tremor. Both in-phase and out-of-phase relationships were observed between TCs. Interestingly, in these six patients, 63/129 single neurons displayed 15-30 Hz oscillations, whereas 36/76 pairs were coherent in this frequency range. Although the oscillatory frequencies were variable between patients, they were highly clustered within a patient. The phase difference between these pairs was found to be close to 0. High-frequency synchronization was observed during periods of limb tremor as well as during intermittent periods with no apparent limb tremor. In contrast, in the three patients without limb tremor during the recording session, only 1/84 neurons had high-frequency oscillatory activity, and no TCs or synchronous high-frequency oscillatory activity was observed (n = 45 pairs). These findings demonstrate that in PD patients with limb tremor, many STN neurons display high-frequency oscillations with a high degree of in-phase synchrony. The results suggest that high-frequency synchronized oscillatory activity may be associated with the pathology that gives rise to tremor in PD patients. PMID- 11027241 TI - Functional magnetic resonance imaging (fMRI) activity in the hippocampal region during recognition memory. AB - Neuroimaging studies have often failed to observe activity in the hippocampal region during memory retrieval. Recently, two functional magnetic resonance imaging studies reported activity in the hippocampal region associated with recollective success. In both, participants studied pictures of objects and were given a recognition memory test with words that either did or did not name the studied objects. The recognition test was therefore cross-modal or associative in nature. These findings raise the question of what circumstances are required to observe activity in the hippocampal region during memory retrieval. Here, we report that robust hippocampal activity for targets relative to foils occurred during retrieval in a recognition memory task when single words were used at both study and test, as well as when pictures of single nameable objects were used at both study and test. The hippocampal region is involved not just in overtly associative tasks but more broadly in the recollection of recently occurring facts and events. PMID- 11027242 TI - A common signaling pathway for striatal NMDA and adenosine A2a receptors: implications for the treatment of Parkinson's disease. AB - The striatum is the major input region of the basal ganglia, playing a pivotal role in the selection, initiation, and coordination of movement both physiologically and in pathophysiological situations such as Parkinson's disease. In the present study, we characterize interactions between NMDA receptors, adenosine receptors, and cAMP signaling within the striatum. Both NMDA (100 micrometer) and the adenosine A(2a) receptor agonist CPCA (3 micrometer) increased cAMP levels (218.9 +/- 19.9% and 395.7 +/- 67.2%, respectively; cf. basal). The NMDA-induced increase in cAMP was completely blocked when slices were preincubated with either the NMDA receptor antagonist 7-chlorokynurenate or the adenosine A(2) receptor antagonist DMPX (100 micrometer), suggesting that striatal NMDA receptors increase cAMP indirectly via stimulation of adenosine A(2a) receptors. Thus, NMDA receptors and adenosine A(2a) receptors might share a common signaling pathway within the striatum. In striatal slices prepared from the 6-hydroxydopamine-lesioned rat model of Parkinson's disease, NMDA receptor mediated increases in cAMP were greater on the lesioned side compared with the unlesioned side (349.6 +/- 40.2% compared with 200.9 +/- 21.9% of basal levels, respectively). This finding substantiates previous evidence implicating overactivity of striatal NMDA receptors in parkinsonism and suggests that a common NMDA receptor-adenosine A(2a) receptor-cAMP signaling cascade might be an important mechanism responsible for mediating parkinsonian symptoms. PMID- 11027243 TI - Effects of irradiance and stimulus duration on early gene expression (Fos) in the suprachiasmatic nucleus: temporal summation and reciprocity. AB - The daily behavioral, physiological, and hormonal rhythms in mammals are regulated by an endogenous circadian clock located in the suprachiasmatic nucleus (SCN) and are synchronized by the natural 24 hr light/dark cycle. We studied the response properties (threshold, saturation, and linearity) of the photic system to irradiance by assaying light induction of Fos, the protein product of the immediate early gene c-fos. Fos expression was quantified by image analysis in the SCN and in the retina. Fos expression in the SCN and retina are unrelated because the response differs in terms of threshold, saturation, and range. In the SCN, Fos expression increases proportionately to increases in both irradiance and duration of light exposure. The photic system shows a linear temporal integration of photons for durations ranging from 3 sec to 47.5 min. The principal result of this study shows that in the SCN, Fos expression is directly proportional to the total number of photons rather than to irradiance or duration alone (reciprocity), and that integration occurs over a range of 5 log units of photon number. This report provides the first demonstration that the mechanism of photon integration by the circadian system is expressed at a cellular level in the SCN. PMID- 11027244 TI - Persistent alterations in dendrites, spines, and dynorphinergic synapses in the nucleus accumbens shell of rats with neuroleptic-induced dyskinesias. AB - Chronic treatment of humans or experimental animals with classical neuroleptic drugs can lead to abnormal, tardive movements that persist long after the drugs are withdrawn. A role in these neuroleptic-induced dyskinesias may be played by a structural change in the shell of the nucleus accumbens where the opioid peptide dynorphin is upregulated in treated rats that show vacuous chewing movements (VCMs). The shell of the nucleus accumbens normally contains a dense plexus of dynorphinergic fibers especially in its caudomedial part. After 27 weeks of haloperidol administration and 18 weeks of withdrawal, the immunoreactive labeling of this plexus is intensified when compared with that after vehicle treatment. In addition, medium spiny neurons here show a significant increase in spine density, dendritic branching, and numbers of terminal segments. In the VCM positive animals, the dendritic surface area is reduced, and dynorphin-positive terminals contact more spines and form more asymmetrical specializations than do those in animals without the syndrome (VCM-negative and vehicle-treated groups). Persistent, neuroleptic-induced oral dyskinesias could therefore be caused by incontrovertible alterations, involving terminal remodeling or sprouting, to the synaptic connectivity of the accumbal shell. PMID- 11027245 TI - Spatial generalization from learning dynamics of reaching movements. AB - When subjects practice reaching movements in a force field, they learn a new sensorimotor map that associates desired trajectories to motor commands. The map is formed in the brain with elements that allow for generalization beyond the region of training. We quantified spatial generalization properties of these elements by training in one extreme of the reachable space and testing near another. Training resulted in rotations in the preferred direction (PD) of activation of some arm muscles. We designed force fields that maintained a constant rotation in muscle PDs as the shoulder joint rotated in the horizontal plane. In such fields, training in a small region resulted in generalization to near and far work spaces (80 cm). In one such field, the forces on the hand reversed directions for a given hand velocity with respect to the location of original training. Despite this, there was generalization. However, if the field was such that the change in the muscle PDs reversed as the work spaces changed, then performance was worse than performance of naive subjects. We suggest that the sensorimotor map of arm dynamics is represented in the brain by elements that globally encode the position of the arm but locally encode its velocity. The elements have preferred directions of movement but are modulated globally by the position of the shoulder joint. We suggest that tuning properties of cells in the motor system influence behavior and that this influence is reflected in the way that we learn dynamics of reaching movements. PMID- 11027246 TI - Stress-level cortisol treatment impairs inhibitory control of behavior in monkeys. AB - Most studies of cortisol-induced cognitive impairments have focused on hippocampal-dependent memory. This study investigates a different aspect of cognition in a randomized placebo-controlled experiment with monkeys that were treated with cortisol according to a protocol that simulates a prolonged stress response. Young adult and older adult monkeys were assigned randomly to placebo or chronic treatment with cortisol in a 2 x 2 factorial design (n = 8 monkeys per condition). Inhibitory control of behavior was assessed with a test shown previously in primates to reflect prefrontal cortical dysfunction. Failure to inhibit a specific goal-directed response was evident more often in older adults. Treatment with cortisol increased this propensity in both older and young adult monkeys. Age-related differences in response inhibition were consistent across blocks of repeated test trials, but the treatment effects were clearly expressed only after prolonged exposure to cortisol. Aspects of performance that did not require inhibition were not altered by age or treatment with cortisol, which concurs with effects on response inhibition rather than nonspecific changes in behavior. These findings lend support to related reports that cortisol-induced disruptions in prefrontal dopamine neurotransmission may contribute to deficits in response inhibition and play a role in cognitive impairments associated with endogenous hypercortisolism in humans. PMID- 11027247 TI - Sensory modification of leech swimming: rhythmic activity of ventral stretch receptors can change intersegmental phase relationships. AB - For segmented animals to generate optimal locomotory movements, appropriate phase relationships between segmental oscillators are crucial. Using swimming leeches, we have investigated the role of sensory input in establishing such relationships. We found that the stretch receptors associated with ventral longitudinal muscles encode the information of muscle contraction during swimming via membrane potential oscillations, with amplitudes of up to 10 mV at our recording site. We subsequently modified the activity of ventral stretch receptors (VSRs) by injecting rhythmic current at different phases of the swim cycle and determined intersegmental phase lags by comparing the delay between the discharges of serially homologous motoneurons in three adjacent segments of isolated nerve cords. When no current was injected, the phase lag between neighboring segments was 8.6 +/- 0.8 degrees (mean +/- SEM; n = 20), with large phase variations from cycle to cycle, between different episodes, and between different preparations. When the phase of stretch receptor activity was set to 90 150 degrees by current injection, the phase of the motoneuron activity in the ganglion was consistently retarded by approximately 5 degrees. It was advanced by approximately 5 degrees when the VSR phase was set to 240-300 degrees. Therefore, the rhythmic activity of the ventral stretch receptor generated during swimming can change intersegmental phase lags of leech ganglia in a phase-dependent manner. These stretch receptors may set the optimal intersegmental phases during swimming movement in intact leeches. PMID- 11027248 TI - Differential cAMP gating of glutamatergic signaling regulates long-term state changes in the suprachiasmatic circadian clock. AB - We investigated a role for cAMP/protein kinase A (PKA) in light/glutamate (GLU) stimulated state changes of the mammalian circadian clock in the suprachiasmatic nucleus (SCN). Nocturnal GLU treatment elevated [cAMP]; however, agonists of cAMP/PKA did not mimic the effects of light/GLU. Coincident activation of cAMP/PKA enhanced GLU-stimulated state changes in early night but blocked light/GLU-induced state changes in the late night, whereas inhibition of cAMP/PKA reversed these effects. These responses are distinct from those mediated by mitogen-activated protein kinase (MAPK). MAPK inhibitors attenuated both GLU induced state changes. Although GLU induced mPer1 mRNA in both early and late night, inhibition of PKA blocked this event only in early night, suggesting that cellular mechanisms regulating mPer1 are gated by the suprachiasmatic circadian clock. These data support a diametric gating role for cAMP/PKA in light/GLU induced SCN state changes: cAMP/PKA promotes the effects of light/GLU in early night, but opposes them in late night. PMID- 11027249 TI - Effect of temperature on dopamine transporter function and intracellular accumulation of methamphetamine: implications for methamphetamine-induced dopaminergic neurotoxicity. AB - Hyperthermia exacerbates and hypothermia attenuates methamphetamine (METH) induced dopamine (DA) neurotoxicity. The mechanisms underlying these temperature effects are unknown. Given the essential role of the DA transporter (DAT) in the expression of METH-induced DA neurotoxicity, we hypothesized that the effect of temperature on METH-induced DA neurotoxicity is mediated, at least in part, at the level of the DAT. To test this hypothesis, the effects of small, physiologically relevant temperature changes on DAT function were evaluated in two types of cultured neuronal cells: (1) a neuroblastoma cell line stably transfected with human DAT cDNA and (2) rat embryonic mesencephalic primary cells that naturally express the DAT. Temperatures for studies of DAT function were selected based on core temperature measurements in animals exposed to METH under usual ambient (22 degrees C) and hypothermic (6 degrees C) temperature conditions, where METH neurotoxicity was fully expressed and blocked, respectively. DAT function, determined by measuring accumulation of radiolabeled DA and 1-methyl-4-phenylpyridinium (MPP(+)), was found to directly correlate with temperature, with higher levels of substrate uptake at 40 degrees C, intermediate levels at 37 degrees C, and lower levels at 34 degrees C. DAT-mediated accumulation of METH also directly correlated with temperature, with greater accumulation at higher temperatures. These findings indicate that relatively small, physiologically relevant changes in temperature significantly alter DAT function and intracellular METH accumulation, and suggest that the effect of temperature on METH-induced DA neurotoxicity is mediated, at least in part, at the level of the DAT. PMID- 11027250 TI - Evidence for spatial modules mediated by temporal synchronization of carbachol induced gamma rhythm in medial entorhinal cortex. AB - Fast (gamma) oscillations in the cortex underlie the rapid temporal coordination of large-scale neuronal assemblies in the processing of sensory stimuli. Cortical gamma rhythm is modulated in vivo by cholinergic innervation from the basal forebrain and can be generated in vitro after exogenous cholinergic stimulation. Using the isolated guinea pig brain, an in vitro preparation that allows for the study of an intact cerebrum, we studied the spatial features of gamma activity evoked by the cholinomimetic carbachol (CCh) in the medial entorhinal cortex (mEC). gamma activity induced by either arterial perfusion or intraparenchymal application of CCh showed a phase reversal across mEC layer II and was reduced or abolished in a spatially localized region by focal infusions of atropine, bicuculline, and CNQX. In addition, a spatially restricted zone of gamma activity could be induced by passive diffusion of CCh from a recording pipette. Finally, gamma oscillations recorded at multiple sites across the surface of the mEC using array electrodes during arterial perfusion of CCh demonstrated a decline in synchronization (coherence) as the interelectrode distance increased. This effect was independent of the signal amplitude and was specific for gamma as opposed to theta-like activity induced by CCh in the same experiments. These results suggest that CCh-induced gamma oscillations in the mEC are mediated through direct muscarinic excitation of a highly localized reciprocal inhibitory-excitatory network located in superficial layers. We propose that functional cortical modules of highly synchronous gamma oscillations may organize incoming (cortical) and outgoing (hippocampal) information in the mEC. PMID- 11027251 TI - Cholinergic inhibition of ventral midbrain dopamine neurons. AB - Muscarinic acetylcholine receptors are common throughout the CNS. The predominant subtypes in the brain are positively coupled to phosphoinositide hydrolysis and have been found to modulate multiple conductances. Muscarinic receptor activation is most often observed to be excitatory because of suppression of various potassium conductances. Here it is reported that three distinct effects of muscarinic receptor activation can be observed in isolation from one another, depending on the duration of receptor activation and the concentration of agonist. Brief activation of muscarinic receptors, as is likely to occur with normal synaptic transmission, hyperpolarized dopamine neurons of the ventral midbrain through a calcium-activated potassium conductance. With repeated or persistent activation of muscarinic receptors, the hyperpolarizing response was entirely desensitized in the absence of any change in resting membrane potential. With sustained activation by higher concentrations of agonist, dopamine neurons were depolarized. This demonstrates that muscarinic receptors can mediate very diverse, and even opposing, postsynaptic effects on neurons depending on the pattern of acetylcholine release. PMID- 11027252 TI - Loading of oxidizable transmitters into secretory vesicles permits carbon-fiber amperometry. AB - Carbon-fiber amperometry detects oxidizable molecules released by exocytosis. We extended this electrochemical technique to cells that do not normally secrete oxidizable transmitters. We incubated AtT-20 cells, pituitary gonadotropes, cultured cerebellar granule cells, and yeast with high concentrations of dopamine (DA) and observed spontaneous and evoked quantal release of DA by amperometry. The rate of detectable spontaneous amperometric events was used as a measure of loading in AtT-20 cells. With 70 mm DA in the bath, loading was complete within 40 min. Cytoplasmic accumulation preceded vesicular loading. Loading decreased proportionally as the bath DA concentration was lowered. Loading rates were similar at 37 and 25 degrees C and much slower at 15 degrees C. Loading was blocked by bafilomycin A(1), a proton pump inhibitor, but not by bupropion, an inhibitor of the plasma membrane DA transporter. Other cells were tested. Spontaneous quantal events became more frequent and evoked events became larger and more frequent when PC12 cells were loaded with DA. Fluid-phase loading of neurons by short stimulation in DA solutions seemed selective for the synaptic vesicles. Thus, many cell types can be loaded with DA to study spontaneous and evoked exocytosis. The amine molecules enter these cells passively and may become concentrated in acidic vesicles by protonation. PMID- 11027253 TI - Stimulation of in vivo dopamine transmission in the bed nucleus of stria terminalis by reinforcing drugs. AB - Drugs of abuse preferentially increase dopamine transmission in the shell of the nucleus accumbens. This area is considered as a transition between the striatum and the extended amygdala a complex neural system that includes the central amygdala and the bed nucleus of stria terminalis, areas that, like the nucleus accumbens shell, are heavily innervated by mesolimbic dopamine neurons originating in the ventral tegmental area. Given the anatomical and neurochemical relationships and similarities with the nucleus accumbens shell it was of interest to investigate whether the dopamine transmission of the bed nucleus of stria terminalis shares with the accumbens shell the peculiar responsiveness to drugs of abuse. To this end we studied by microdialysis with concentric probes, the effect of drugs of abuse on extracellular dopamine in the bed nucleus of stria terminalis. We report that morphine, nicotine, cocaine, ethanol, and the selective dopamine uptake inhibitor GBR 12909 increase effectively and dose dependently extracellular dopamine in the bed nucleus of stria terminalis. These results indicate that the bed nucleus of stria terminalis shares with the nucleus accumbens shell a peculiar sensitivity to the dopamine stimulant actions of drugs of abuse. PMID- 11027254 TI - Inositol 1,4,5-triphosphate-evoked responses in midbrain dopamine neurons. AB - Synaptically released glutamate evokes slow IPSPs mediated by metabotropic glutamate receptors (mGluRs) in midbrain dopamine neurons. These mGluR IPSPs are caused by release of Ca(2+) from intracellular stores and subsequent activation of small-conductance Ca(2+)-activated K(+) channels (SK channels). To further investigate the intracellular mechanisms involved, the effect of photolyzing intracellular caged inositol 1,4,5-triphosphate (InsP(3)) on membrane conductance and intracellular Ca(2+) concentration ([Ca(2+)](i)) was examined in rat midbrain slices. Photolytic release of InsP(3) elicited a transient outward current and a sharp rise in [Ca(2+)](i) that lasted for approximately 5 sec. Apamin, a blocker of SK channels, abolished the InsP(3)-induced outward current without affecting the rise in [Ca(2+)](i). Depleting intracellular Ca(2+) stores with cyclopiazonic acid completely blocked both the outward current and the Ca(2+) transient elicited by InsP(3). InsP(3)-evoked Ca(2+) mobilization was not affected by blockade of ryanodine receptors with ruthenium red, whereas depleting ryanodine sensitive Ca(2+) stores with ryanodine almost eliminated InsP(3)-induced Ca(2+) release. Increasing the size of intracellular Ca(2+) stores by means of prolonged depolarization added a late component to the outward current and a slow component to the rising phase of [Ca(2+)](i). These effects of depolarization were blocked by ruthenium red. These results show that InsP(3) activates SK channels by releasing Ca(2+) from InsP(3)-sensitive stores that also contain ryanodine receptors. Increasing intracellular Ca(2+) stores boosts InsP(3)-evoked responses by invoking Ca(2+)-induced Ca(2+) release through ryanodine receptors. This intracellular signaling pathway may play a significant role in regulating the excitability of midbrain dopamine neurons. PMID- 11027255 TI - Roles for mismatch repair factors in regulating genetic recombination. PMID- 11027256 TI - The abundance of Met30p limits SCF(Met30p) complex activity and is regulated by methionine availability. AB - Ubiquitin-mediated degradation plays a crucial role in many fundamental biological pathways, including the mediation of cellular responses to changes in environmental conditions. A family of ubiquitin ligase complexes, called SCF complexes, found throughout eukaryotes, is involved in a variety of biological pathways. In Saccharomyces cerevisiae, an SCF complex contains a common set of components, namely, Cdc53p, Skp1p, and Hrt1p. Substrate specificity is defined by a variable component called an F-box protein. The F- box is a approximately 40 amino-acid motif that allows the F-box protein to bind Skp1p. Each SCF complex recognizes different substrates according to which F-box protein is associated with the complex. In yeasts, three SCF complexes have been demonstrated to associate with the ubiquitin-conjugating enzyme Cdc34p and have ubiquitin ligase activity. F-box proteins are not abundant and are unstable. As part of the SCF(Met30p) complex, the F-box protein Met30p represses methionine biosynthetic gene expression when availability of L-methionine is high. Here we demonstrate that in vivo SCF(Met30p) complex activity can be regulated by the abundance of Met30p. Furthermore, we provide evidence that Met30p abundance is regulated by the availability of L-methionine. We propose that the cellular responses mediated by an SCF complex are directly regulated by environmental conditions through the control of F-box protein stability. PMID- 11027257 TI - Analysis of fission yeast primase defines the checkpoint responses to aberrant S phase initiation. AB - To investigate the checkpoint response to aberrant initiation, we analyzed the cell cycle checkpoint response induced by mutations of Schizosaccharomyces pombe DNA primase. DNA primase has two subunits, Spp1 and Spp2 (S. pombe primases 1 and 2). Spp1 is the catalytic subunit that synthesizes the RNA primer, which is then extended by DNA polymerase alpha (Polalpha) to synthesize an initiation DNA structure, and this catalytic function of Polalpha is a prerequisite for generating the S-M phase checkpoint. Here we show that Spp2 is required for coupling the function of Spp1 to Polalpha. Thermosensitive mutations of spp2(+) destabilize the Polalpha-primase complex, resulting in an allele-specific S phase checkpoint defect. The mutant exhibiting a more severe checkpoint defect also has a higher extent of Polalpha-primase complex instability and deficiency in the hydroxyurea-induced Cds1-mediated intra-S phase checkpoint response. However, this mutant is able to activate the Cds1 response to S phase arrest induced by temperature. These findings suggest that the Cds1 response to the S-phase arrest signal(s) induced by a initiation mutant is different from that induced by hydroxyurea. Interestingly, a polalphats mutant with a defective S-M phase checkpoint and an spp2 mutant with an intact checkpoint have a similar Polalpha primase complex stability, and the Cds1 response induced by hydroxyurea or by the mutant arrests at the restrictive temperature. Thus, the Cds1-mediated intra-S phase checkpoint response induced by hydroxyurea can also be distinguished from the S-M phase checkpoint response that requires the initiation DNA synthesis by Polalpha. PMID- 11027258 TI - Nckbeta adapter regulates actin polymerization in NIH 3T3 fibroblasts in response to platelet-derived growth factor bb. AB - The SH3-SH3-SH3-SH2 adapter Nck represents a two-gene family that includes Nckalpha (Nck) and Nckbeta (Grb4/Nck2), and it links receptor tyrosine kinases to intracellular signaling networks. The function of these mammalian Nck genes has not been established. We report here a specific role for Nckbeta in platelet derived growth factor (PDGF)-induced actin polymerization in NIH 3T3 cells. Overexpression of Nckbeta but not Nckalpha blocks PDGF-stimulated membrane ruffling and formation of lamellipoda. Mutation in either the SH2 or the middle SH3 domain of Nckbeta abolishes its interfering effect. Nckbeta binds at Tyr-1009 in human PDGF receptor beta (PDGFR-beta) which is different from Nckalpha's binding site, Tyr-751, and does not compete with phosphatidylinositol-3 kinase for binding to PDGFR. Microinjection of an anti-Nckbeta but not an anti-Nckalpha antibody inhibits PDGF-stimulated actin polymerization. Constitutively membrane bound Nckbeta but not Nckalpha blocks Rac1-L62-induced membrane ruffling and formation of lamellipodia, suggesting that Nckbeta acts in parallel to or downstream of Rac1. This is the first report of Nckbeta's role in receptor tyrosine kinase signaling to the actin cytoskeleton. PMID- 11027259 TI - In yeast, the 3' untranslated region or the presequence of ATM1 is required for the exclusive localization of its mRNA to the vicinity of mitochondria. AB - We isolated mitochondria from Saccharomyces cerevisiae to selectively study polysomes bound to the mitochondrial surface. The distribution of several mRNAs coding for mitochondrial proteins was examined in free and mitochondrion-bound polysomes. Some mRNAs exclusively localize to mitochondrion-bound polysomes, such as the ones coding for Atm1p, Cox10p, Tim44p, Atp2p, and Cot1p. In contrast, mRNAs encoding Cox6p, Cox5a, Aac1p, and Mir1p are found enriched in free cytoplasmic polysome fractions. Aac1p and Mir1p are transporters that lack cleavable presequences. Sequences required for mRNA asymmetric subcellular distribution were determined by analyzing the localization of reporter mRNAs containing the presequence coding region and/or the 3'-untranslated region (3'UTR) of ATM1, a gene encoding an ABC transporter of the mitochondrial inner membrane. Biochemical analyses of mitochondrion-bound polysomes and direct visualization of RNA localization in living yeast cells allowed us to demonstrate that either the presequence coding region or the 3'UTR of ATM1 is sufficient to allow the reporter mRNA to localize to the vicinity of the mitochondrion, independently of its translation. These data demonstrate that mRNA localization is one of the mechanisms used, in yeast, for segregating mitochondrial proteins. PMID- 11027260 TI - Saccharomyces cerevisiae expresses three functionally distinct homologues of the nramp family of metal transporters. AB - The baker's yeast Saccharomyces cerevisiae expresses three homologues of the Nramp family of metal transporters: Smf1p, Smf2p, and Smf3p, encoded by SMF1, SMF2, and SMF3, respectively. Here we report a comparative analysis of the yeast Smf proteins at the levels of localization, regulation, and function of the corresponding metal transporters. Smf1p and Smf2p function in cellular accumulation of manganese, and the two proteins are coregulated by manganese ions and the BSD2 gene product. Under manganese-replete conditions, Bsd2p facilitates trafficking of Smf1p and Smf2p to the vacuole, where these transport proteins are degraded. However, Smf1p and Smf2p localize to distinct cellular compartments under metal starvation: Smf1p accumulates at the cell surface, while Smf2p is restricted to intracellular vesicles. The third Nramp homologue, Smf3p, is quite distinctive. Smf3p is not regulated by Bsd2p or by manganese ions and is not degraded in the vacuole. Instead, Smf3p is down-regulated by iron through a mechanism that does not involve transcription or protein stability. Smf3p localizes to the vacuolar membrane independently of metal treatment, and yeast cells lacking Smf3p show symptoms of iron starvation. We propose that Smf3p helps to mobilize vacuolar stores of iron. PMID- 11027261 TI - Down-regulation of cyclin D1 expression by prostaglandin A(2) is mediated by enhanced cyclin D1 mRNA turnover. AB - Prostaglandin A(2) (PGA(2)), an experimental chemotherapeutic agent, causes growth arrest associated with decreased cyclin D1 expression in several cancer cell lines. Here, using human non-small-cell lung carcinoma H1299 cells, we investigated the mechanisms whereby PGA(2) down-regulates cyclin D1 expression. Transcription rates of the cyclin D1 gene, studied using a cyclin D1 promoter luciferase construct and nuclear run-on assays, were not affected by PGA(2) treatment. Instead, the cyclin D1 mRNA was rendered unstable after exposure to PGA(2). Since the stability of labile mRNA is modulated through binding of proteins to specific mRNA sequences, we sought to identify protein(s) recognizing the cyclin D1 mRNA. In electrophoretic mobility-shift assays using radiolabeled RNA probes derived from different regions of cyclin D1 mRNA, we observed that (i) lysates prepared from PGA(2)-treated cells exhibited enhanced protein-cyclin D1 RNA complex formation; (ii) the kinetics of complex formation correlated closely with that of cyclin D1 mRNA loss; and (iii) binding occurred within a 390-base cyclin D1 3' untranslated region (UTR) (K12). This binding activity could be cross-linked, revealing proteins ranging from 30 to 47 kDa. The RNA-binding protein AUF1, previously associated with the degradation of target mRNAs, bound cyclin D1 mRNA, because anti-AUF1 antibodies were capable of supershifting or immunoprecipitating cyclin D1 mRNA-protein complexes. Finally, insertion of K12 in the 3'UTR of reporter genes markedly reduced the expression and half-life of the resulting chimeric mRNAs in transfected, PGA(2)-treated cells. Our data demonstrate that PGA(2) down-regulates cyclin D1 expression by decreasing cyclin D1 mRNA stability and implicates a 390-base element in the 3'UTR in this regulation. PMID- 11027262 TI - The nicking step in V(D)J recombination is independent of synapsis: implications for the immune repertoire. AB - In all of the transposition reactions that have been characterized thus far, synapsis of two transposon ends is required before any catalytic steps (strand nicking or strand transfer) occur. In V(D)J recombination, there have been inconclusive data concerning the role of synapsis in nicking. Synapsis between two 12-substrates or between two 23-substrates has not been ruled out in any studies thus far. Here we provide the first direct tests of this issue. We find that immobilization of signals does not affect their nicking, even though hairpinning is affected in a manner reflecting its known synaptic requirement. We also find that nicking is kinetically a unireactant enzyme-catalyzed reaction. Time courses are no different between nicking seen for a 12-substrate alone and a reaction involving both a 12- and a 23-substrate. Hence, synapsis is neither a requirement nor an effector of the rate of nicking. These results establish V(D)J recombination as the first example of a DNA transposition-type reaction in which catalytic steps begin prior to synapsis, and the results have direct implications for the order of the steps in V(D)J recombination, for the contribution of V(D)J recombination nicks to genomic instability, and for the diversification of the immune repertoire. PMID- 11027263 TI - Schizosaccharomyces pombe Hsk1p is a potential cds1p target required for genome integrity. AB - The fission yeast Hsk1p kinase is an essential activator of DNA replication. Here we report the isolation and characterization of a novel mutant allele of the gene. Consistent with its role in the initiation of DNA synthesis, hsk1(ts) genetically interacts with several S-phase mutants. At the restrictive temperature, hsk1(ts) cells suffer abnormal S phase and loss of nuclear integrity and are sensitive to both DNA-damaging agents and replication arrest. Interestingly, hsk1(ts) mutants released to the restrictive temperature after early S-phase arrest in hydroxyurea (HU) are able to complete bulk DNA synthesis but they nevertheless undergo an abnormal mitosis. These findings indicate a second role for hsk1 subsequent to HU arrest. Consistent with a later S-phase role, hsk1(ts) is synthetically lethal with Deltarqh1 (RecQ helicase) or rad21ts (cohesin) mutants and suppressed by Deltacds1 (RAD53 kinase) mutants. We demonstrate that Hsk1p undergoes Cds1p-dependent phosphorylation in response to HU and that it is a direct substrate of purified Cds1p kinase in vitro. These results indicate that the Hsk1p kinase is a potential target of Cds1p regulation and that its activity is required after replication initiation for normal mitosis. PMID- 11027264 TI - mRNA decapping in yeast requires dissociation of the cap binding protein, eukaryotic translation initiation factor 4E. AB - A major pathway of eukaryotic mRNA turnover occurs by deadenylation-dependent decapping that exposes the transcript to 5'-->3' exonucleolytic degradation. A critical step in this pathway is decapping, since removal of the cap structure permits 5'-->3' exonucleolytic digestion. Based on alterations in mRNA decay rate from strains deficient in translation initiation, it has been proposed that the decapping rate is modulated by a competition between the cytoplasmic cap binding complex, which promotes translation initiation, and the decapping enzyme, Dcp1p. In order to test this model directly, we examined the functional interaction of Dcp1p and the cap binding protein, eukaryotic translation initiation factor 4E (eIF4E), in vitro. These experiments indicated that eIF4E is an inhibitor of Dcp1p in vitro due to its ability to bind the 5' cap structure. In addition, we demonstrate that in vivo a temperature-sensitive allele of eIF4E (cdc33-42) suppressed the decapping defect of a partial loss-of-function allele of DCP1. These results argue that dissociation of eIF4E from the cap structure is required before decapping. Interestingly, the temperature-sensitive allele of eIF4E does not suppress the decapping defect seen in strains lacking the decapping activators, Lsm1p and Pat1p. This indicates that these activators of decapping affect a step in mRNA turnover distinct from the competition between Dcp1 and eIF4E. PMID- 11027265 TI - Functional characterization of nuclear localization signals in yeast Sm proteins. AB - In mammals, nuclear localization of U-snRNP particles requires the snRNA hypermethylated cap structure and the Sm core complex. The nature of the signal located within the Sm core proteins is still unknown, both in humans and yeast. Close examination of the sequences of the yeast SmB, SmD1, and SmD3 carboxyl terminal domains reveals the presence of basic regions that are reminiscent of nuclear localization signals (NLSs). Fluorescence microscopy studies using green fluorescent protein (GFP)-fusion proteins indicate that both yeast SmB and SmD1 basic amino acid stretches exhibit nuclear localization properties. Accordingly, deletions or mutations in the NLS-like motifs of SmB and SmD1 dramatically reduce nuclear fluorescence of the GFP-Sm mutant fusion alleles. Phenotypic analyses indicate that the NLS-like motifs of SmB and SmD1 are functionally redundant: each NLS-like motif can be deleted without affecting yeast viability whereas a simultaneous deletion of both NLS-like motifs is lethal. Taken together, these findings suggest that, in the doughnut-like structure formed by the Sm core complex, the carboxyl-terminal extensions of Sm proteins may form an evolutionarily conserved basic amino acid-rich protuberance that functions as a nuclear localization determinant. PMID- 11027266 TI - Evidence for splice site pairing via intron definition in Schizosaccharomyces pombe. AB - Schizosaccharomyces pombe pre-mRNAs are generally multi-intronic and share certain features with pre-mRNAs from Drosophila melanogaster, in which initial splice site pairing can occur via either exon or intron definition. Here, we present three lines of evidence suggesting that, despite these similarities, fission yeast splicing is most likely restricted to intron definition. First, mutating either or both splice sites flanking an internal exon in the S. pombe cdc2 gene produced almost exclusively intron retention, in contrast to the exon skipping observed in vertebrates. Second, we were unable to induce skipping of the internal microexon in fission yeast cgs2, whereas the default splicing pathway excludes extremely small exons in mammals. Because nearly quantitative removal of the downstream intron in cgs2 could be achieved by expanding the microexon, we propose that its retention is due to steric occlusion. Third, several cryptic 5' junctions in the second intron of fission yeast cdc2 are located within the intron, in contrast to their generally exonic locations in metazoa. The effects of expanding and contracting this intron are as predicted by intron definition; in fact, even highly deviant 5' junctions can compete effectively with the standard 5' splice site if they are closer to the 3' splicing signals. Taken together, our data suggest that pairing of splice sites in S. pombe most likely occurs exclusively across introns in a manner that favors excision of the smallest segment possible. PMID- 11027267 TI - Yeast Krr1p physically and functionally interacts with a novel essential Kri1p, and both proteins are required for 40S ribosome biogenesis in the nucleolus. AB - Using a two-hybrid screening with TOM1, a putative ubiquitin-ligase gene of Saccharomyces cerevisiae, we isolated KRR1, a homologue of human HRB2 (for human immunodeficiency virus type 1 Rev-binding protein 2). To characterize the gene function, we constructed temperature-sensitive krr1 mutants and isolated two multicopy suppressors. One suppressor is RPS14A, encoding a 40S ribosomal protein. The C-terminal-truncated rpS14p, which was reported to have diminished binding activity to 18S rRNA, failed to suppress the krr1 mutant. The other suppressor is a novel gene, KRI1 (for KRR1 interacting protein; YNL308c). KRI1 is essential for viability, and Kri1p is localized to the nucleolus. We constructed a galactose-dependent kri1 strain by placing KRI1 under control of the GAL1 promoter, so that expression of KRI1 was shut off when transferring the culture to glucose medium. Polysome and 40S ribosome fractions were severely decreased in the krr1 mutant and Kri1p-depleted cells. Pulse-chase analysis of newly synthesized rRNAs demonstrated that 18S rRNA is not produced in either mutant. However, wild-type levels of 25S rRNA are made in either mutant. Northern analysis revealed that the steady-state levels of 18S rRNA and 20S pre-rRNAs were reduced in both mutants. Precursors for 18S rRNA were detected but probably very unstable in both mutants. A myc-tagged Kri1p coimmunoprecipitated with a hemagglutinin-tagged Krr1p. Furthermore, the krr1 mutant protein was defective in its interaction with Kri1p. These data lead us to conclude that Krr1p physically and functionally interacts with Kri1p to form a complex which is required for 40S ribosome biogenesis in the nucleolus. PMID- 11027268 TI - Defining the roles of nucleotide excision repair and recombination in the repair of DNA interstrand cross-links in mammalian cells. AB - The mechanisms by which DNA interstrand cross-links (ICLs) are repaired in mammalian cells are unclear. Studies in bacteria and yeasts indicate that both nucleotide excision repair (NER) and recombination are required for their removal and that double-strand breaks are produced as repair intermediates in yeast cells. The role of NER and recombination in the repair of ICLs induced by nitrogen mustard (HN2) was investigated using Chinese hamster ovary mutant cell lines. XPF and ERCC1 mutants (defective in genes required for NER and some types of recombination) and XRCC2 and XRCC3 mutants (defective in RAD51-related homologous recombination genes) were highly sensitive to HN2. Cell lines defective in other genes involved in NER (XPB, XPD, and XPG), together with a mutant defective in nonhomologous end joining (XRCC5), showed only mild sensitivity. In agreement with their extreme sensitivity, the XPF and ERCC1 mutants were defective in the incision or "unhooking" step of ICL repair. In contrast, the other mutants defective in NER activities, the XRCC2 and XRCC3 mutants, and the XRCC5 mutant all showed normal unhooking kinetics. Using pulsed field gel electrophoresis, DNA double-strand breaks (DSBs) were found to be induced following nitrogen mustard treatment. DSB induction and repair were normal in all the NER mutants, including XPF and ERCC1. The XRCC2, XRCC3, and XRCC5 mutants also showed normal induction kinetics. The XRCC2 and XRCC3 homologous recombination mutants were, however, severely impaired in the repair of DSBs. These results define a role for XPF and ERCC1 in the excision of ICLs, but not in the recombinational components of cross-link repair. In addition, homologous recombination but not nonhomologous end joining appears to play an important role in the repair of DSBs resulting from nitrogen mustard treatment. PMID- 11027269 TI - Telomere folding is required for the stable maintenance of telomere position effects in yeast. AB - Yeast telomeres reversibly repress the transcription of adjacent genes, a phenomenon called telomere position effect (TPE). TPE is thought to result from Rap1 and Sir protein-mediated spreading of heterochromatin-like structures from the telomeric DNA inwards. Because Rap1p is associated with subtelomeric chromatin as well as with telomeric DNA, yeast telomeres are proposed to form fold-back or looped structures. TPE can be eliminated in trans by deleting SIR genes or in cis by transcribing through the C(1-3)A/TG(1-3) tract of a telomere. We show that the promoter of a telomere-linked URA3 gene was inaccessible to restriction enzymes and that accessibility increased both in a sir3 strain and upon telomere transcription. We also show that subtelomeric chromatin was hypoacetylated at histone H3 and at each of the four acetylatable lysines in histone H4 and that histone acetylation increased both in a sir3 strain and when the telomere was transcribed. When transcription through the telomeric tract occurred in G(1)-arrested cells, TPE was lost, demonstrating that activation of a silenced telomeric gene can occur in the absence of DNA replication. The loss of TPE that accompanied telomere transcription resulted in the rapid and efficient loss of subtelomeric Rap1p. We propose that telomere transcription disrupts core heterochromatin by eliminating Rap1p-mediated telomere looping. This interpretation suggests that telomere looping is critical for maintaining TPE. PMID- 11027270 TI - Replication past O(6)-methylguanine by yeast and human DNA polymerase eta. AB - O(6)-Methylguanine (m6G) is formed by the action of alkylating agents such as N methyl-N'-nitro-N-nitrosoguanidine (MNNG) on DNA. m6G is a highly mutagenic and carcinogenic lesion, and it presents a block to synthesis by DNA polymerases. Here, we provide genetic and biochemical evidence for the involvement of yeast and human DNA polymerase eta (Poleta) in the replicative bypass of m6G lesions in DNA. The formation of MNNG-induced mutations is almost abolished in the rad30Delta pol32Delta double mutant of yeast, which lacks the RAD30 gene that encodes Poleta and the Pol32 subunit of DNA polymerase delta (Poldelta). Although Poldelta can function in the mutagenic bypass of m6G lesions, our biochemical studies indicate that Poleta is much more efficient in replicating through m6G than Poldelta. Both Poleta and Poldelta insert a C or a T residue opposite from m6G; Poleta, however, is more accurate, as it inserts a C about twice as frequently as Poldelta. Alkylating agents are used in the treatment of malignant tumors, including lymphomas, brain tumors, melanomas, and gastrointestinal carcinomas, and the clinical effectiveness of these agents derives at least in part from their ability to form m6G in DNA. Inactivation of Poleta could afford a useful strategy for enhancing the effectiveness of these agents in cancer chemotherapy. PMID- 11027271 TI - Discrete roles for peroxisome proliferator-activated receptor gamma and retinoid X receptor in recruiting nuclear receptor coactivators. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a major role in adipogenesis. PPARgamma binds to DNA as a heterodimer with retinoid X receptor (RXR), and PPARgamma-RXR can be activated by ligands specific for either receptor; the presence of both ligands can result in a cooperative effect on the transactivation of target genes. How these ligands mediate transactivation, however, remains unclear. PPARgamma is known to interact with both the p160/SRC-1 family of coactivators and the distinct, multisubunit coactivator complex called DRIP. A single DRIP subunit, DRIP205 (TRAP220, PBP), binds directly to PPARgamma. Here we report that PPARgamma and RXR selectively interacted with DRIP205 and p160 proteins in a ligand-dependent manner. At physiological concentrations, RXR specific ligands only induced p160 binding to RXR, and PPARgamma-specific ligands exclusively recruited DRIP205 but not p160 coactivators to PPARgamma. This selectivity was not observed in interaction assays off DNA, implying that the specificity of coactivator binding in response to ligand is strongly influenced by the allosteric effects of DNA-bound heterodimers. These coactivator-selective effects were also observed in transient-transfection assays in the presence of overexpressed p160 or DRIP coactivators. The results suggest that the cooperative effects of PPARgamma- and RXR-specific ligands may occur at the level of selective coactivator recruitment. PMID- 11027272 TI - Proapoptotic p53-interacting protein 53BP2 is induced by UV irradiation but suppressed by p53. AB - p53 is an important mediator of the cellular stress response with roles in cell cycle control, DNA repair, and apoptosis. 53BP2, a p53-interacting protein, enhances p53 transactivation, impedes cell cycle progression, and promotes apoptosis through unknown mechanisms. We now demonstrate that endogenous 53BP2 levels increase following UV irradiation induced DNA damage in a p53-independent manner. In contrast, we found that the presence of a wild-type (but not mutant) p53 gene suppressed 53BP2 steady-state levels in cell lines with defined p53 genotypes. Likewise, expression of a tetracycline-regulated wild-type p53 cDNA in p53-null fibroblasts caused a reduction in 53BP2 protein levels. However, 53BP2 levels were not reduced if the tetracycline-regulated p53 cDNA was expressed after UV damage in these cells. This suggests that UV damage activates cellular factors that can relieve the p53-mediated suppression of 53BP2 protein. To address the physiologic significance of 53BP2 induction, we utilized stable cell lines with a ponasterone A-regulated 53BP2 cDNA. Conditional expression of 53BP2 cDNA lowered the apoptotic threshold and decreased clonogenic survival following UV irradiation. Conversely, attenuation of endogenous 53BP2 induction with an antisense oligonucleotide resulted in enhanced clonogenic survival following UV irradiation. These results demonstrate that 53BP2 is a DNA damage-inducible protein that promotes DNA damage-induced apoptosis. Furthermore, 53BP2 expression is highly regulated and involves both p53-dependent and p53-independent mechanisms. Our data provide new insight into 53BP2 function and open new avenues for investigation into the cellular response to genotoxic stress. PMID- 11027273 TI - ets-2 is a target for an akt (Protein kinase B)/jun N-terminal kinase signaling pathway in macrophages of motheaten-viable mutant mice. AB - The transcription factor ets-2 was phosphorylated at residue threonine 72 in a colony-stimulating factor 1 (CSF-1)- and mitogen-activated protein kinase independent manner in macrophages isolated from motheaten-viable (me-v) mice. The CSF-1 and ets-2 target genes coding for Bcl-x, urokinase plasminogen activator, and scavenger receptor were also expressed at high levels independent of CSF-1 addition to me-v cells. Akt (protein kinase B) was constitutively active in me-v macrophages, and an Akt immunoprecipitate catalyzed phosphorylation of ets-2 at threonine 72. The p54 isoform of c-jun N-terminal kinase-stress-activated kinase (JNK- SAPK) coimmunoprecipitated with Akt from me-v macrophages, and treatment of me-v cells with the specific phosphatidylinositol 3-kinase inhibitor LY294002 decreased cell survival, Akt and JNK kinase activities, ets-2 phosphorylation, and Bcl-x mRNA expression. Therefore, ets-2 is a target for phosphatidylinositol 3-kinase-Akt-JNK action, and the JNK p54 isoform is an ets-2 kinase in macrophages. Constitutive ets-2 activity may contribute to the pathology of me-v mice by increasing expression of genes like the Bcl-x gene that promote macrophage survival. PMID- 11027274 TI - Positive and negative regulation of phosphoinositide 3-kinase-dependent signaling pathways by three different gene products of the p85alpha regulatory subunit. AB - Phosphoinositide (PI) 3-kinase is a key mediator of insulin-dependent metabolic actions, including stimulation of glucose transport and glycogen synthesis. The gene for the p85alpha regulatory subunit yields three splicing variants, p85alpha, AS53/p55alpha, and p50alpha. All three have (i) a C-terminal structure consisting of two Src homology 2 domains flanking the p110 catalytic subunit binding domain and (ii) a unique N-terminal region of 304, 34, and 6 amino acids, respectively. To determine if these regulatory subunits differ in their effects on enzyme activity and signal transduction from insulin receptor substrate (IRS) proteins under physiological conditions, we expressed each regulatory subunit in fully differentiated L6 myotubes using adenovirus-mediated gene transfer with or without coexpression of the p110alpha catalytic subunit. PI 3-kinase activity associated with p50alpha was greater than that associated with p85alpha or AS53. Increasing the level of p85alpha or AS53, but not p50alpha, inhibited both phosphotyrosine-associated and p110-associated PI 3-kinase activities. Expression of a p85alpha mutant lacking the p110-binding site (Deltap85) also inhibited phosphotyrosine-associated PI 3-kinase activity but not p110-associated activity. Insulin stimulation of two kinases downstream from PI-3 kinase, Akt and p70 S6 kinase (p70(S6K)), was decreased in cells expressing p85alpha or AS53 but not in cells expressing p50alpha. Similar inhibition of PI 3-kinase, Akt, and p70(S6K) was observed, even when p110alpha was coexpressed with p85alpha or AS53. Expression of p110alpha alone dramatically increased glucose transport but decreased glycogen synthase activity. This effect was reduced when p110alpha was coexpressed with any of the three regulatory subunits. Thus, the three different isoforms of regulatory subunit can relay the signal from IRS proteins to the p110 catalytic subunit with different efficiencies. They also negatively modulate the PI 3-kinase catalytic activity but to different extents, dependent on the unique N-terminal structure of each isoform. These data also suggest the existence of a mechanism by which regulatory subunits modulate the PI 3-kinase-mediated signals, independent of the kinase activity, possibly through subcellular localization of the catalytic subunit or interaction with additional signaling molecules. PMID- 11027275 TI - Identification of novel Saccharomyces cerevisiae proteins with nuclear export activity: cell cycle-regulated transcription factor ace2p shows cell cycle independent nucleocytoplasmic shuttling. AB - Nuclear export of proteins containing leucine-rich nuclear export signals (NESs) is mediated by the NES receptor CRM1/Crm1p. We have carried out a yeast two hybrid screen with Crm1p as a bait. The Crm1p-interacting clones were subscreened for nuclear export activity in a visual assay utilizing the Crm1p-inhibitor leptomycin B (LMB). This approach identified three Saccharomyces cerevisiae proteins not previously known to have nuclear export activity. These proteins are the 5' RNA triphosphatase Ctl1p, the cell cycle-regulated transcription factor Ace2p, and a protein encoded by the previously uncharacterized open reading frame YDR499W. Mutagenesis analysis show that YDR499Wp contains an NES that conforms to the consensus sequence for leucine-rich NESs. Mutagenesis of Ctl1p and Ace2p were unable to identify specific NES residues. However, a 29-amino-acid region of Ace2p, rich in hydrophobic residues, contains nuclear export activity. Ace2p accumulates in the nucleus at the end of mitosis and activates early-G(1) specific genes. We now provide evidence that Ace2p is nuclear not only in late M early G(1) but also during other stages of the cell cycle. This feature of Ace2p localization explains its ability to activate genes such as CUP1, which are not expressed in a cell cycle-dependent manner. PMID- 11027276 TI - Recombinogenic flap ligation pathway for intrinsic repair of topoisomerase IB induced double-strand breaks. AB - Topoisomerase IB catalyzes recombinogenic DNA strand transfer reactions in vitro and in vivo. Here we characterize a new pathway of topoisomerase-mediated DNA ligation in vitro (flap ligation) in which vaccinia virus topoisomerase bound to a blunt-end DNA joins the covalently held strand to a 5' resected end of a duplex DNA containing a 3' tail. The joining reaction occurs with high efficiency when the sequence of the 3' tail is complementary to that of the scissile strand immediately 5' of the cleavage site. A 6-nucleotide segment of complementarity suffices for efficient flap ligation. Invasion of the flap into the duplex apparently occurs while topoisomerase remains bound to DNA, thereby implying a conformational flexibility of the topoisomerase clamp around the DNA target site. The 3' flap acceptor DNA mimics a processed end in the double-strand-break-repair recombination pathway. Our findings suggest that topoisomerase-induced breaks may be rectified by flap ligation, with ensuing genomic deletions or translocations. PMID- 11027277 TI - Role of phosphoinositide 3-kinase and endocytosis in nerve growth factor-induced extracellular signal-regulated kinase activation via Ras and Rap1. AB - Neurotrophins promote multiple actions on neuronal cells including cell survival and differentiation. The best-studied neurotrophin, nerve growth factor (NGF), is a major survival factor in sympathetic and sensory neurons and promotes differentiation in a well-studied model system, PC12 cells. To mediate these actions, NGF binds to the TrkA receptor to trigger intracellular signaling cascades. Two kinases whose activities mediate these processes include the mitogen-activated protein (MAP) kinase (or extracellular signal-regulated kinase [ERK]) and phosphoinositide 3-kinase (PI3-K). To examine potential interactions between the ERK and PI3-K pathways, we studied the requirement of PI3-K for NGF activation of the ERK signaling cascade in dorsal root ganglion cells and PC12 cells. We show that PI3-K is required for TrkA internalization and participates in NGF signaling to ERKs via distinct actions on the small G proteins Ras and Rap1. In PC12 cells, NGF activates Ras and Rap1 to elicit the rapid and sustained activation of ERKs respectively. We show here that Rap1 activation requires both TrkA internalization and PI3-K, whereas Ras activation requires neither TrkA internalization nor PI3-K. Both inhibitors of PI3-K and inhibitors of endocytosis prevent GTP loading of Rap1 and block sustained ERK activation by NGF. PI3-K and endocytosis may also regulate ERK signaling at a second site downstream of Ras, since both rapid ERK activation and the Ras-dependent activation of the MAP kinase kinase kinase B-Raf are blocked by inhibition of either PI3-K or endocytosis. The results of this study suggest that PI3-K may be required for the signals initiated by TrkA internalization and demonstrate that specific endocytic events may distinguish ERK signaling via Rap1 and Ras. PMID- 11027278 TI - Ral GTPases contribute to regulation of cyclin D1 through activation of NF kappaB. AB - Ral GTPases have been implicated as mediators of Ras-induced signal transduction from observations that Ral-specific guanine nucleotide exchange factors associate with Ras and are activated by Ras. The cellular role of Ral family proteins is unclear, as is the contribution that Ral may make to Ras-dependent signaling. Here we show that expression of activated Ral in quiescent rodent fibroblasts is sufficient to induce activation of NF-kappaB-dependent gene expression and cyclin D1 transcription, two key convergence points for mitogenic and survival signaling. The regulation of cyclin D1 transcription by Ral is dependent on NF kappaB activation and is mediated through an NF-kappaB binding site in the cyclin D1 promoter. Ral activation of these responses is likely through an as yet uncharacterized effector pathway, as we find activation of NF-kappaB and the cyclin D1 promoter by Ral is independent of association of Ral with active phospholipase D1 or Ral-binding protein 1, two proteins proposed to mediate Ral function in cells. PMID- 11027279 TI - Tryptophan permease gene TAT2 confers high-pressure growth in Saccharomyces cerevisiae. AB - Hydrostatic pressure in the range of 15 to 25 MPa was found to cause arrest of the cell cycle in G(1) phase in an exponentially growing culture of Saccharomyces cerevisiae, whereas a pressure of 50 MPa did not. We found that a plasmid carrying the TAT2 gene, which encodes a high-affinity tryptophan permease, enabled the cells to grow under conditions of pressure in the range of 15 to 25 MPa. Additionally, cells expressing the Tat2 protein at high levels became endowed with the ability to grow under low-temperature conditions at 10 or 15 degrees C as well as at high pressure. Hydrostatic pressure significantly inhibited tryptophan uptake into the cells, and the Tat2 protein level was down regulated by high pressure. The activation volume associated with tryptophan uptake was found to be a large positive value, 46.2 +/- 3.85 ml/mol, indicating that there was a net volume increase in a rate-limiting step in tryptophan import. The results showing cell cycle arrest in G(1) phase and down-regulation of the Tat2 protein seem to be similar to those observed upon treatment of cells with the immunosuppressive drug rapamycin. Although rapamycin treatment elicited the rapid dephosphorylation of Npr1 and induction of Gap1 expression, hydrostatic pressure did not affect the phosphorylation state of Npr1 and it decreased the level of Gap1 protein, suggesting that the pressure-sensing pathway may be independent of Npr1 function. Here we describe high-pressure sensing in yeast in comparison with the TOR-signaling pathway and discuss an important factor involved in adaptation of organisms to high-pressure environments. PMID- 11027280 TI - Inactivation of smad-transforming growth factor beta signaling by Ca(2+) calmodulin-dependent protein kinase II. AB - Members of the transforming growth factor beta (TGF-beta) family transduce signals through Smad proteins. Smad signaling can be regulated by the Ras/Erk/mitogen-activated protein pathway in response to receptor tyrosine kinase activation and the gamma interferon pathway and also by the functional interaction of Smad2 with Ca(2+)-calmodulin. Here we report that Smad-TGF-beta dependent transcriptional responses are prevented by expression of a constitutively activated Ca(2+)-calmodulin-dependent protein kinase II (Cam kinase II). Smad2 is a target substrate for Cam kinase II in vitro at serine-110, -240, and -260. Cam kinase II induces in vivo phosphorylation of Smad2 and Smad4 and, to a lesser extent, Smad3. A phosphopeptide antiserum raised against Smad2 phosphoserine-240 reacted with Smad2 in vivo when coexpressed with Cam kinase II and by activation of the platelet-derived growth factor receptor, the epidermal growth factor receptor, HER2 (c-erbB2), and the TGF-beta receptor. Furthermore, Cam kinase II blocked nuclear accumulation of a Smad2 and induced Smad2-Smad4 hetero-oligomerization independently of TGF-beta receptor activation, while preventing TGF-beta-dependent Smad2-Smad3 interactions. These findings provide a novel cross-talk mechanism by which Ca(2+)-dependent kinases activated downstream of multiple growth factor receptors antagonize cell responses to TGF-beta. PMID- 11027281 TI - Target selectivity of bicoid is dependent on nonconsensus site recognition and protein-protein interaction. AB - We describe experiments to compare the activities of two Drosophila homeodomain proteins, Bicoid (Bcd) and an altered-specificity mutant of Fushi tarazu, Ftz(Q50K). Although the homeodomains of these proteins share a virtually indistinguishable ability to recognize a consensus Bcd site, only Bcd can activate transcription from natural enhancer elements when assayed in both yeast and Drosophila Schneider S2 cells. Our analysis of chimeric proteins suggests that both the homeodomain of Bcd and sequences outside the homeodomain contribute to its ability to recognize natural enhancer elements. We further show that, unlike the Bcd homeodomain, the Ftz(Q50K) homeodomain fails to recognize nonconsensus sites found in natural enhancer elements. The defect of a chimeric protein containing the homeodomain of Ftz(Q50K) in place of that of Bcd can be preferentially restored by converting the nonconsensus sites in natural enhancer elements to consensus sites. Our experiments suggest that the biological specificity of Bcd is determined by combinatorial contributions of two important mechanisms: the nonconsensus site recognition function conferred by the homeodomain and the cooperativity function conferred primarily by sequences outside the homeodomain. A systematic comparison of different assay methods and enhancer elements further suggests a fluid nature of the requirements for these two Bcd functions in target selection. PMID- 11027282 TI - Multiple mechanisms of suppression circumvent transcription defects in an RNA polymerase mutant. AB - Using a high-copy-number suppressor screen to obtain clues about the role of the yeast RNA polymerase II subunit RPB4 in transcription, we identified three suppressors of the temperature sensitivity resulting from deletion of the RPB4 gene (DeltaRPB4). One suppressor is Sro9p, a protein related to La protein, another is the nucleosporin Nsp1p, and the third is the RNA polymerase II subunit RPB7. Suppression by RPB7 was anticipated since its interaction with RPB4 is well established both in vitro and in vivo. We examined the effect of overexpression of each suppressor gene on transcription. Interestingly, suppression of the temperature-sensitive phenotype correlates with the correction of a characteristic transcription defect of this mutant: each suppressor restored the level of promoter-specific, basal transcription to wild-type levels. Examination of the effects of the suppressors on other in vivo transcription aberrations in DeltaRPB4 cells revealed significant amelioration of defects in certain inducible genes in Sro9p and RPB7, but not in Nsp1p, suppressor cells. Analysis of mRNA levels demonstrated that overexpression of each of the three suppressors minimally doubled the mRNA levels during stationary phase. However, the elevated mRNA levels in Sro9p suppressor cells appear to result from a combination of enhanced transcription and message stability. Taken together, these results demonstrate that these three proteins influence transcription and implicate Sro9p in both transcription and posttranscription events. PMID- 11027283 TI - cis- and trans-Acting determinants for translation of psbD mRNA in Chlamydomonas reinhardtii. AB - Chloroplast translation is mediated by nucleus-encoded factors that interact with distinct cis-acting RNA elements. A U-rich sequence within the 5' untranslated region of the psbD mRNA has previously been shown to be required for its translation in Chlamydomonas reinhardtii. By using UV cross-linking assays, we have identified a 40-kDa RNA binding protein, which binds to the wild-type psbD leader, but is unable to recognize a nonfunctional leader mutant lacking the U rich motif. RNA binding is restored in a chloroplast cis-acting suppressor. The functions of several site-directed psbD leader mutants were analyzed with transgenic C. reinhardtii chloroplasts and the in vitro RNA binding assay. A clear correlation between photosynthetic activity and the capability to bind RNA by the 40-kDa protein was observed. Furthermore, the data obtained suggest that the poly(U) region serves as a molecular spacer between two previously characterized cis-acting elements, which are involved in RNA stabilization and translation. RNA-protein complex formation depends on the nuclear Nac2 gene product that is part of a protein complex required for the stabilization of the psbD mRNA. The sedimentation properties of the 40-kDa RNA binding protein suggest that it interacts directly with this Nac2 complex and, as a result, links processes of chloroplast RNA metabolism and translation. PMID- 11027284 TI - Loss of a protein phosphatase 2A regulatory subunit (Cdc55p) elicits improper regulation of Swe1p degradation. AB - CDC55 encodes a Saccharomyces cerevisiae protein phosphatase 2A (PP2A) regulatory subunit. cdc55-null cells growing at low temperature exhibit a failure of cytokinesis and produce abnormally elongated buds, but cdc55-null cells producing the cyclin-dependent kinase Cdc28-Y19F, which is unable to be inhibited by Y19 phosphorylation, show a loss of the abnormal morphology. Furthermore, cdc55-null cells exhibit a hyperphosphorylation of Y19. For these reasons, we have examined in wild-type and cdc55-null cells the levels and activities of the kinase (Swe1p) and phosphatase (Mih1p) that normally regulate the extent of Cdc28 Y19 phosphorylation. We find that Mih1p levels are comparable in the two strains, and an estimate of the in vivo and in vitro phosphatase activity of this enzyme in the two cell types indicates no marked differences. By contrast, while Swe1p levels are similar in unsynchronized and S-phase-arrested wild-type and cdc55 null cells, Swe1 kinase is found at elevated levels in mitosis-arrested cdc55 null cells. This excess Swe1p in cdc55-null cells is the result of ectopic stabilization of this protein during G(2) and M, thereby accounting for the accumulation of Swe1p in mitosis-arrested cells. We also present evidence indicating that, in cdc55-null cells, misregulated PP2A phosphatase activity is the cause of both the ectopic stabilization of Swe1p and the production of the morphologically abnormal phenotype. PMID- 11027285 TI - Regulatory networks revealed by transcriptional profiling of damaged Saccharomyces cerevisiae cells: Rpn4 links base excision repair with proteasomes. AB - Exposure to carcinogenic alkylating agents, oxidizing agents, and ionizing radiation modulates transcript levels for over one third of Saccharomyces cerevisiae's 6,200 genes. Computational analysis delineates groups of coregulated genes whose upstream regions bear known and novel regulatory sequence motifs. One group of coregulated genes contain a number of DNA excision repair genes (including the MAG1 3-methyladenine DNA glycosylase gene) and a large selection of protein degradation genes. Moreover, transcription of these genes is modulated by the proteasome-associated protein Rpn4, most likely via its binding to MAG1 upstream repressor sequence 2-like elements, that turn out to be almost identical to the recently identified proteasome-associated control element (G. Mannhaupt, R. Schnall, V. Karpov, I. Vetter, and H. Feldmann, FEBS Lett. 450:27-34, 1999). We have identified a large number of genes whose transcription is influenced by Rpn4p. PMID- 11027286 TI - Mechanism of promoter melting by the xeroderma pigmentosum complementation group B helicase of transcription factor IIH revealed by protein-DNA photo-cross linking. AB - The p89/xeroderma pigmentosum complementation group B (XPB) ATPase-helicase of transcription factor IIH (TFIIH) is essential for promoter melting prior to transcription initiation by RNA polymerase II (RNAPII). By studying the topological organization of the initiation complex using site-specific protein DNA photo-cross-linking, we have shown that p89/XPB makes promoter contacts both upstream and downstream of the initiation site. The upstream contact, which is in the region where promoter melting occurs (positions -9 to +2), requires tight DNA wrapping around RNAPII. The addition of hydrolyzable ATP tethers the template strand at positions -5 and +1 to RNAPII subunits. A mutation in p89/XPB found in a xeroderma pigmentosum patient impairs the ability of TFIIH to associate correctly with the complex and thereby melt promoter DNA. A model for open complex formation is proposed. PMID- 11027287 TI - Telomerase-associated protein TEP1 is not essential for telomerase activity or telomere length maintenance in vivo. AB - TEP1 is a mammalian telomerase-associated protein with similarity to the Tetrahymena telomerase protein p80. Like p80, TEP1 is associated with telomerase activity and the telomerase reverse transcriptase, and it specifically interacts with the telomerase RNA. To determine the role of mTep1 in telomerase function in vivo, we generated mouse embryonic stem (ES) cells and mice lacking mTep1. The mTep1-deficient (mTep1(-/-)) mice were viable and were bred for seven successive generations with no obvious phenotypic abnormalities. All murine tissues from mTep1(-/-) mice possessed a level of telomerase activity comparable to that in wild-type mice. In addition, analysis of several tissues that normally lack telomerase activity revealed no reactivation of telomerase activity in mTep1(-/-) mice. Telomere length, even in later generations of mTep1(-/-) mice, was equivalent to that in wild-type animals. ES cells deficient in mTep1 also showed no detectable alteration in telomerase activity or telomere length with increased passage in culture. Thus, mTep1 appears to be completely dispensable for telomerase function in vivo. Recently, TEP1 has been identified within a second ribonucleoprotein (RNP) complex, the vault particle. TEP1 can also specifically bind to a small RNA, vRNA, which is associated with the vault particle and is unrelated in sequence to mammalian telomerase RNA. These results reveal that TEP1 is an RNA binding protein that is not restricted to the telomerase complex and that TEP1 plays a redundant role in the assembly or localization of the telomerase RNP in vivo. PMID- 11027288 TI - The CUL1 C-terminal sequence and ROC1 are required for efficient nuclear accumulation, NEDD8 modification, and ubiquitin ligase activity of CUL1. AB - Members of the cullin and RING finger ROC protein families form heterodimeric complexes to constitute a potentially large number of distinct E3 ubiquitin ligases. We report here that the highly conserved C-terminal sequence in CUL1 is dually required, both for nuclear localization and for modification by NEDD8. Disruption of ROC1 binding impaired nuclear accumulation of CUL1 and decreased NEDD8 modification in vivo but had no effect on NEDD8 modification of CUL1 in vitro, suggesting that ROC1 promotes CUL1 nuclear accumulation to facilitate its NEDD8 modification. Disruption of NEDD8 binding had no effect on ROC1 binding, nor did it affect nuclear localization of CUL1, suggesting that nuclear localization and NEDD8 modification of CUL1 are two separable steps, with nuclear import preceding and required for NEDD8 modification. Disrupting NEDD8 modification diminishes the IkappaBalpha ubiquitin ligase activity of CUL1. These results identify a pathway for regulation of CUL1 activity-ROC1 and the CUL1 C terminal sequence collaboratively mediate nuclear accumulation and NEDD8 modification, facilitating assembly of active CUL1 ubiquitin ligase. This pathway may be commonly utilized for the assembly of other cullin ligases. PMID- 11027289 TI - Arginine-rich regions mediate the RNA binding and regulatory activities of the protein encoded by the Drosophila melanogaster suppressor of sable gene. AB - The Drosophila melanogaster suppressor of sable gene, su(s), encodes a novel, 150 kDa nuclear RNA binding protein, SU(S), that negatively regulates RNA accumulation from mutant alleles of other genes that have transposon insertions in the 5' transcribed region. In this study, we delineated the RNA binding domain of SU(S) and evaluated its relevance to SU(S) function in vivo. As a result, we have defined two arginine-rich motifs (ARM1 and ARM2) that mediate the RNA binding activity of SU(S). ARM1 is required for in vitro high-affinity binding of SU(S) to small RNAs that were previously isolated by SELEX (binding site selection assay) and that contain a common consensus sequence. ARM1 is also required for the association of SU(S) with larval polytene chromosomes in vivo. ARM2 promotes binding of SU(S) to SELEX RNAs that lack the consensus sequence and apparently is neither necessary nor sufficient for the stable polytene chromosome association of SU(S). Use of the GAL4/UAS system to drive ectopic expression of su(s) cDNA transgenes revealed two previously unknown properties of SU(S). First, overexpression of SU(S) is lethal. Second, SU(S) negatively regulates expression of su(s) intronless cDNA transgenes, and the ARMs are required for this effect. Considering these and previous results, we propose that SU(S) binds to the 5' region of nascent transcripts and inhibits RNA production in a manner that can be overcome by splicing complex assembly. PMID- 11027290 TI - Profilin II is alternatively spliced, resulting in profilin isoforms that are differentially expressed and have distinct biochemical properties. AB - We deduced the structure of the mouse profilin II gene. It contains five exons that can generate four different transcripts by alternative splicing. Two transcripts encode different profilin II isoforms (designated IIa and IIb) that have similar affinities for actin but different affinities for polyphosphoinositides and proline-rich sequences. Profilins IIa and IIb are also present in humans, suggesting that all mammals have three profilin isoforms. Profilin I is the major form in all tissues, except in the brain, where profilin IIa is most abundant. Profilin IIb appears to be a minor form, and its expression is restricted to a limited number of tissues, indicating that the alternative splicing is tightly regulated. Western blotting and whole-mount in situ hybridization show that, in contrast to the expression of profilin I, the expression level of profilin IIa is developmentally regulated. In situ hybridization of adult brain sections reveals overlapping expression patterns of profilins I and IIa. PMID- 11027291 TI - Roughex mediates G(1) arrest through a physical association with cyclin A. AB - Differentiation in the developing Drosophila eye requires synchronization of cells in the G(1) phase of the cell cycle. The roughex gene product plays a key role in this synchronization by negatively regulating cyclin A protein levels in G(1). We show here that coexpressed Roughex and cyclin A physically interact in vivo. Roughex is a nuclear protein, while cyclin A was previously shown to be exclusively cytoplasmic during interphase in the embryo. In contrast, we demonstrate that in interphase cells in the eye imaginal disk cyclin A is present in both the nucleus and the cytoplasm. In the presence of ectopic Roughex, cyclin A becomes strictly nuclear and is later degraded. Nuclear targeting of both Roughex and cyclin A under these conditions is dependent on a C-terminal nuclear localization signal in Roughex. Disruption of this signal results in cytoplasmic localization of both Roughex and cyclin A, confirming a physical interaction between these molecules. Cyclin A interacts with both Cdc2 and Cdc2c, the Drosophila Cdk2 homolog, and Roughex inhibits the histone H1 kinase activities of both cyclin A-Cdc2 and cyclin A-Cdc2c complexes in whole-cell extracts. Two hybrid experiments suggested that the inhibition of kinase activity by Roughex results from competition with the cyclin-dependent kinase subunit for binding to cyclin A. These findings suggest that Roughex can influence the intracellular distribution of cyclin A and define Roughex as a distinct and specialized cell cycle inhibitor for cyclin A-dependent kinase activity. PMID- 11027292 TI - Function of the ski4p (Csl4p) and Ski7p proteins in 3'-to-5' degradation of mRNA. AB - One of two general pathways of mRNA decay in the yeast Saccharomyces cerevisiae occurs by deadenylation followed by 3'-to-5' degradation of the mRNA body. Previous results have shown that this degradation requires components of the exosome and the Ski2p, Ski3p, and Ski8p proteins, which were originally identified due to their superkiller phenotype. In this work, we demonstrate that deletion of the SKI7 gene, which encodes a putative GTPase, also causes a defect in 3'-to-5' degradation of mRNA. Deletion of SKI7, like deletion of SKI2, SKI3, or SKI8, does not affect various RNA-processing reactions of the exosome. In addition, we show that a mutation in the SKI4 gene also causes a defect in 3'-to 5' mRNA degradation. We show that the SKI4 gene is identical to the CSL4 gene, which encodes a core component of the exosome. Interestingly, the ski4-1 allele contains a point mutation resulting in a mutation in the putative RNA binding domain of the Csl4p protein. This point mutation strongly affects mRNA degradation without affecting exosome function in rRNA or snRNA processing, 5' externally transcribed spacer (ETS) degradation, or viability. In contrast, the csl4-1 allele of the same gene affects rRNA processing but not 3'-to-5' mRNA degradation. We identify csl4-1 as resulting from a partial-loss-of-function mutation in the promoter of the CSL4 gene. These data indicate that the distinct functions of the exosome can be separated genetically and suggest that the RNA binding domain of Csl4p may have a specific function in mRNA degradation. PMID- 11027293 TI - Human scribble (Vartul) is targeted for ubiquitin-mediated degradation by the high-risk papillomavirus E6 proteins and the E6AP ubiquitin-protein ligase. AB - The high-risk human papillomavirus (HPV) E6 proteins stimulate the ubiquitination and degradation of p53, dependent on the E6AP ubiquitin-protein ligase. Other proteins have also been shown to be targeted for degradation by E6, including hDlg, the human homolog of the Drosophila melanogaster Discs large (Dlg) tumor suppressor. We show here that the human homolog of the Drosophila Scribble (Vartul) (hScrib) tumor suppressor protein is also targeted for ubiquitination by the E6-E6AP complex in vitro and that expression of E6 induces degradation of hScrib in vivo. Characterization of the E6AP-E6-hScrib complex indicated that hScrib binds directly to E6 and that the binding is mediated by the PDZ domains of hScrib and a carboxyl-terminal epitope conserved among the high-risk HPV E6 proteins. Green fluorescent protein-hScrib was localized to the periphery of MDCK cells, where it colocalized with ZO-1, a component of tight junctions. E6 expression resulted in loss of integrity of tight junctions, as measured by ZO-1 localization, and this effect was dependent on the PDZ binding epitope of E6. Thus, the high-risk HPV E6 proteins induce the degradation of the human homologs of two Drosophila PDZ domain-containing tumor suppressor proteins, hDlg and hScrib, both of which are associated with cell junction complexes. The fact that Scrib/Vart and Dlg appear to cooperate in a pathway that controls Drosophila epithelial cell growth suggests that the combined targeting of hScrib and hDlg is an important component of the biologic activity of high-risk HPV E6 proteins. PMID- 11027294 TI - Inhibition of p300 histone acetyltransferase by viral interferon regulatory factor. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) has been consistently identified in Kaposi's sarcomas, body cavity-based lymphomas, and some forms of Castleman's disease. The K9 open reading frame of KSHV encodes a viral interferon regulatory factor (vIRF) which functions as a repressor for cellular interferon-mediated signal transduction and as an oncogene to induce cell growth transformation. We demonstrate that KSHV vIRF directly interacts with cellular transcriptional coactivator p300 and displaces p300/CBP-associated factor from p300 complexes. This interaction inhibits the histone acetyltransferase activity of p300, resulting in drastic reduction of nucleosomal histone acetylation and alteration of chromatin structure. As a consequence, vIRF expression markedly alters cellular cytokine expression, which is regulated by acetylation of nucleosomal histones. These results demonstrate that KSHV vIRF interacts with and inhibits the p300 transcriptional coactivator to circumvent the host antiviral immune response and to induce a global alteration of cellular gene expression. These studies also illustrate how a cellular gene captured by a herpesvirus has evolved several functions that suit the needs of the virus. PMID- 11027296 TI - DNA replication is required To elicit cellular responses to psoralen-induced DNA interstrand cross-links. AB - Following introduction of DNA interstrand cross-links (ICLs), mammalian cells display chromosome breakage or cell cycle delay with a 4N DNA content. To further understand the nature of the delay, previously described as a G(2)/M arrest, we developed a protocol to generate ICLs during specific intervals of the cell cycle. Synchronous populations of G(1), S, and G(2) cells were treated with photoactivated 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) and scored for normal passage into mitosis. In contrast to what was found for ionizing radiation, ICLs introduced during G(2) did not result in a G(2)/M arrest, mitotic arrest, or chromosome breakage. Rather, subsequent passage through S phase was required to trigger both chromosome breakage and arrest in the next cell cycle. Similarly, ICLs introduced during G(1) did not cause a G(1)/S arrest. We conclude that DNA replication is required to elicit the cellular responses of cell cycle arrest and genomic instability after psoralen-induced ICLs. In primary human fibroblasts, the 4N DNA content cell cycle arrest triggered by ICLs was long lasting but reversible. Kinetic analysis suggested that these cells could remove up to approximately 2,500 ICLs/genome at an average rate of 11 ICLs/genome/h. PMID- 11027295 TI - Elevated levels of hepatocyte nuclear factor 3beta in mouse hepatocytes influence expression of genes involved in bile acid and glucose homeostasis. AB - The winged helix transcription factor, hepatocyte nuclear factor-3beta (HNF 3beta), mediates the hepatocyte-specific transcription of numerous genes important for liver function. However, the in vivo role of HNF-3beta in regulating these genes remains unknown because homozygous null HNF3beta mouse embryos die in utero prior to liver formation. In order to examine the regulatory function of HNF-3beta, we created transgenic mice in which the -3-kb transthyretin promoter functions to increase hepatocyte expression of the rat HNF 3beta protein. Postnatal transgenic mice exhibit growth retardation, depletion of hepatocyte glycogen storage, and elevated levels of bile acids in serum. The retarded growth phenotype is likely due to a 20-fold increase in hepatic expression of insulin-like growth factor binding protein 1 (IGFBP-1), which results in elevated levels in serum of IGFBP-1 and limits the biological availability of IGFs required for postnatal growth. The defects in glycogen storage and serum bile acids coincide with diminished postnatal expression of hepatocyte genes involved in gluconeogenesis (phosphoenolpyruvate carboxykinase and glycogen synthase) and sinusoidal bile acid uptake (Ntcp), respectively. These changes in gene transcription may result from the disruptive effect of HNF 3beta on the hepatic expression of the endogenous mouse HNF-3alpha,-3beta, 3gamma, and -6 transcription factors. Furthermore, adult transgenic livers lack expression of the canalicular phospholipid transporter, mdr2, which is consistent with ultrastructure evidence of damage to transgenic hepatocytes and bile canaliculi. These transgenic studies represent the first in vivo demonstration that the HNF-3beta transcriptional network regulates expression of hepatocyte specific genes required for bile acid and glucose homeostasis, as well as postnatal growth. PMID- 11027297 TI - Participation of the C-terminal domain of RNA polymerase II in exon definition during pre-mRNA splicing. AB - Interaction between transcription and pre-mRNA processing via binding of polymerase II (Pol II) to factors involved in capping, splicing, and polyadenylation has recently been demonstrated. The C-terminal domain (CTD), a highly phosphorylated repeat sequence of the largest subunit of Pol II, has been implicated in this interaction because deletion of this domain affects downstream RNA processing events and because it is the binding site for numerous processing factors. Here we show that recombinant CTD, free of other components of Pol II, activated in vitro splicing and assembly of the spliceosome in nuclear extracts if, and only if, the assayed precursor RNA was recognized via exon definition, i.e., if the substrates contained complete exons with both 3' and 5' splice sites. Furthermore, depletion of intact Pol II inactivated splicing of this set of precursor RNAs and addition of recombinant CTD restored activity. The added recombinant CTD was quickly hyper- and hypophosphorylated in extract, became associated with the precursor RNA, and stimulated the association of U1 snRNPs but not ASF/SF2 with substrate RNA. These observations suggest that the mode of interaction between the CTD and splicing factors is integrally tied to exon definition and the mechanism whereby distal exons can be recognized and brought into juxtaposition during assembly of the spliceosome. PMID- 11027298 TI - Allosteric, chiral-selective drug binding to DNA. AB - The binding interactions of (-)-daunorubicin (WP900), a newly synthesized enantiomer of the anticancer drug (+)-daunorubicin, with right- and left-handed DNA, have been studied quantitatively by equilibrium dialysis, fluorescence spectroscopy, and circular dichroism. (+)-Daunorubicin binds selectively to right handed DNA, whereas the enantiomeric WP900 ligand binds selectively to left handed DNA. Further, binding of the enantiomeric pair to DNA is clearly chirally selective, and each of the enantiomers was found to act as an allosteric effector of DNA conformation. Under solution conditions that initially favored the left handed conformation of [poly(dGdC)](2), (+)-daunorubicin allosterically converted the polynucleotide to a right-handed intercalated form. In contrast, under solution conditions that initially favored the right-handed conformation of [poly(dGdC)](2), WP900 converted the polynucleotide to a left-handed form. Molecular dynamics studies by using the amber force field resulted in a stereochemically feasible model for the intercalation of WP900 into left-handed DNA. The chiral selectivity observed for the DNA binding of the daunorubicin/WP900 enantiomeric pair is far greater than the selectivity previously reported for a variety of chiral metal complexes. These results open a new avenue for the rational design of potential anticancer agents that target left-handed DNA. PMID- 11027299 TI - A conserved mechanism of retrovirus restriction in mammals. AB - The murine Fv1 gene restricts infection by N- or B-tropic murine leukemia viruses at a postentry, preintegration stage. The Fv1-sensitive viruses previously used for the study of Fv1 encode an ecotropic envelope gene and thus only infect rodent cells. Consequently, the study of Fv1 restriction has been carried out solely in mice and murine cell lines. By infection with retroviral vectors containing N- or B-tropic core and pantropic vesicular stomatitis virus-G envelope protein, we now demonstrate that cell lines derived from various mammalian species, including humans, have an Fv1-like retrovirus restriction function, preventing N-tropic vector infection. Like Fv1, restriction is directed at amino acid 110 of the viral capsid protein. In contrast to Fv1, the novel restriction is characterized by the absence of reverse-transcribed viral DNA. We speculate that these activities have been selected for by retroviral epidemics in the distant past. PMID- 11027300 TI - BCR/ABL inhibition by an escort/phosphatase fusion protein. AB - Cellular transformation by the BCR/ABL oncogene depends on the ABL-encoded tyrosine kinase activity. To block BCR/ABL function, we created a unique tyrosine phosphatase by fusing the catalytic domain of SHP1 (SHP1c) to the ABL binding domain (ABD) of RIN1, an established binding partner and substrate for c-ABL and BCR/ABL. This fusion construct (ABD/SHP1c) binds to BCR/ABL in cells and functions as an active phosphatase. ABD/SHP1c effectively suppressed BCR/ABL function as judged by reductions in transformation of fibroblast cells, growth factor independence of hematopoietic cell lines, and proliferation of primary bone marrow cells. In addition, the leukemogenic properties of BCR/ABL in a murine model system were blocked by coexpression of ABD/SHP1c. Both the "escort" function provided by ABD and the inhibitor function provided by the phosphatase of SHP1c were necessary for effective BCR/ABL interference. Expression of ABD/SHP1c also reversed the transformed phenotype of K562, a human leukemia derived cell line. These results have direct implications for leukemia therapeutics and suggest an approach to block aberrant signal transduction in other pathologies through the use of appropriately designed escort/inhibitors. PMID- 11027301 TI - Substrate-dependent mutant complementation to select fatty acid desaturase variants for metabolic engineering of plant seed oils. AB - We demonstrate that naturally occurring C(14) and C(16)-specific acyl-acyl carrier protein (ACP) desaturases from plants can complement the unsaturated fatty acid (UFA) auxotrophy of an Escherichia coli fabA/fadR mutant. Under the same growth conditions, C(18)-specific delta(9)-stearoyl (18:0)-ACP desaturases are unable to complement the UFA auxotrophy. This difference most likely results from the presence of sufficient substrate pools of C(14) and C(16) acyl-ACPs but a relative lack of C(18) acyl-ACP pools in E. coli to support the activities of the plant fatty acid desaturase. Based on this, a substrate-dependent selection system was devised with the use of the E. coli UFA auxotroph to isolate mutants of the castor delta(9)-18:0-ACP desaturase that display enhanced specificity for C(14) and C(16) acyl-ACPs. Using this selection system, a number of desaturase variants with altered substrate specificities were isolated from pools of randomized mutants. These included several G188L mutant isolates, which displayed a 15-fold increase in specific activity with 16:0-ACP relative to the wild-type castor delta(9)-18:0-ACP desaturase. Expression of this mutant in Arabidopsis thaliana resulted in the accumulation of unusual monounsaturated fatty acids to amounts of >25% of the seed oil. The bacterial selection system described here thus provides a rapid means of isolating variant fatty acid desaturase activities for modification of seed oil composition. PMID- 11027302 TI - Negative hair-bundle stiffness betrays a mechanism for mechanical amplification by the hair cell. AB - Hearing and balance rely on the ability of hair cells in the inner ear to sense miniscule mechanical stimuli. In each cell, sound or acceleration deflects the mechanosensitive hair bundle, a tuft of rigid stereocilia protruding from the cell's apical surface. By altering the tension in gating springs linked to mechanically sensitive transduction channels, this deflection changes the channels' open probability and elicits an electrical response. To detect weak stimuli despite energy losses caused by viscous dissipation, a hair cell can use active hair-bundle movement to amplify its mechanical inputs. This amplificatory process also yields spontaneous bundle oscillations. Using a displacement-clamp system to measure the mechanical properties of individual hair bundles from the bullfrog's ear, we found that an oscillatory bundle displays negative slope stiffness at the heart of its region of mechanosensitivity. Offsetting the hair bundle's position activates an adaptation process that shifts the region of negative stiffness along the displacement axis. Modeling indicates that the interplay between negative bundle stiffness and the motor responsible for mechanical adaptation produces bundle oscillation similar to that observed. Just as the negative resistance of electrically excitable cells and of tunnel diodes can be embedded in a biasing circuit to amplify electrical signals, negative stiffness can be harnessed to amplify mechanical stimuli in the ear. PMID- 11027303 TI - Demonstration of the in vivo interaction of key cell death regulators by structure-based design of second-site suppressors. AB - Demonstrating in vivo interaction of two important biomolecules and the relevance of the interaction to a biological process have been difficult issues in biomedical research. Here, we report the use of a homology modeling approach to establish the significance of protein interactions in governing the activation of programmed cell death in Caenorhabditis elegans. A protein interaction cascade has been postulated to mediate activation of cell death in nematodes, in which the BH3-domain-containing (Bcl-2 homology region 3) protein EGL-1 binds the cell death inhibitor CED-9 and induces release of the death-activating protein CED-4 from inhibitory CED-4/CED-9 complexes. We show here that an unusual gain-of function mutation in ced-9 (substitution of glycine 169 to glutamate) that results in potent inhibition of most nematode cell deaths impairs the binding of EGL-1 to CED-9 and EGL-1-induced release of CED-4 from CED-4/CED-9 complexes. Based on a modeled EGL-1/CED-9 complex structure, we generated second-site compensatory mutations in EGL-1 that partially restore the binding of EGL-1 to CED-9(G169E) and EGL-1-induced release of CED-4 from CED-4/CED-9(G169E) complexes. Importantly, these mutations also significantly suppress the death protective activity of CED-9(G169E) in vivo. These results establish that direct physical interaction between EGL-1 and CED-9 is essential for the release of CED 4 and the activation of cell death. The structure-based design of second-site suppressors via homology modeling should be widely applicable for probing important molecular interactions that are implicated in fundamental biological processes. PMID- 11027304 TI - Prostitution and the sex discrepancy in reported number of sexual partners. AB - One of the most reliable and perplexing findings from surveys of sexual behavior is that men report substantially more sexual partners than women do. We use data from national sex surveys and studies of prostitutes and their clients in the United States to examine sampling bias as an explanation for this disparity. We find that prostitute women are underrepresented in the national surveys. Once their undersampling and very high numbers of sexual partners are factored in, the discrepancy disappears. Prostitution's role in the discrepancy is not readily apparent because men are reluctant to acknowledge that their reported partners include prostitutes. PMID- 11027305 TI - Sulindac inhibits neointimal formation after arterial injury in wild-type and apolipoprotein E-deficient mice. AB - Neointimal hyperplasia is a critical component of restenosis, a major complication of angioplasty and related therapeutic procedures. We studied the effects of hyperlipidemia and the nonsteroidal anti-inflammatory drugs, aspirin (acetyl-salicylic acid; ASA), and sulindac, on neointimal formation in a mouse femoral arterial injury model. At 2 months of age, normolipidemic, wild-type (WT), and hyperlipidemic, apolipoprotein E-deficient (apoE-/-) mice were divided into three treatment groups: Western-type diet (WD), WD + ASA (200 mg/kg food), and WD + sulindac (300 mg/kg food). After 1 week, mice underwent arterial injury and treatments were maintained for 4 weeks. Histomorphometry of the injured arteries showed striking effects of plasma cholesterol levels and drug treatment on neointimal hyperplasia. In the WD or WD + ASA groups, apoE-/- mice had twice the neointimal area than WT mice ( approximately 30,000 vs. 13,000 microm(2) per section; P < 0.0001). Compared with ASA or WD alone, sulindac treatment resulted in approximately 70% (P = 0.0001) and 50% (P = 0.01) reductions in the neointimal area in apoE-/- and WT mice, respectively. ASA, at a dose sufficient to inhibit platelet aggregation, did not affect neointimal formation in mice of either genotype. Evidence of macrophages was noted in the lesions of apoE-/- mice in the WD and WD + ASA groups, but remarkably, none was detectable with sulindac treatment, despite hyperlipidemia, suggesting early steps in the response to injury were abrogated. These results demonstrate sulindac reduces neointimal formation in both normolipidemic and hyperlipidemic settings and raise the possibility that similar benefits may be obtained in patients undergoing angioplasty and related procedures. PMID- 11027306 TI - Functional switching of GABAergic synapses by ryanodine receptor activation. AB - The role of the ryanodine receptor (RyR) in modifiability of synapses made by the basket interneurons onto the hippocampal CA1 pyramidal cells was examined in rats. Associating single-cell RyR activation with postsynaptic depolarization increased intracellular free Ca(2+) concentrations and reversed the basket interneuron-CA1 inhibitory postsynaptic potential into an excitatory postsynaptic potential. This synaptic transformation was accompanied by a shift of the reversal potential from that of chloride toward that of bicarbonate. This inhibitory postsynaptic potential-excitatory postsynaptic potential transformation was prevented by blocking RyR or carbonic anhydrase. Associated postsynaptic depolarization and RyR activation, therefore, changes GABAergic synapses from excitation filters to amplifier and, thereby, shapes information flow through the hippocampal network. PMID- 11027307 TI - Receptor-mediated endocytosis of soluble and membrane-tethered Sonic hedgehog by Patched-1. AB - Patched (Ptc) is the ligand-binding component of the Hedgehog (Hh) receptor complex. In the Drosophila embryo, Ptc and Hh colocalize in vesicular punctate structures. However, receptor-mediated endocytosis of Hh proteins has not been demonstrated. By using chick neural plate explants, we show that Sonic hedgehog (Shh)-responsive neural precursor cells internalize recombinant and endogenous Shh and provide direct evidence for a gradient of endogenous Shh in the ventral neural tube. Shh internalization is blocked by a monoclonal antibody whose epitope overlaps the Ptc-binding site of Shh. These findings suggest that Shh internalization is mediated by Ptc-1 and may be linked to signaling. Concordantly, transfection of mammalian cell lines with a Ptc-1 cDNA confers the ability to internalize multiple forms of Shh, including transmembrane-anchored Shh, by a dynamin-dependent process. PMID- 11027308 TI - Dual roles of the 11S regulatory subcomplex in condensin functions. AB - Condensin is a multisubunit protein complex that reconfigures DNA structure in an ATP-dependent manner in vitro and plays a central role in mitotic chromosome condensation in Xenopus egg cell-free extracts. The Xenopus 13S condensin complex (13SC) is composed of two subcomplexes: an 8S core subcomplex (8SC) consisting of two structural maintenance of chromosomes (SMC) subunits (XCAP-C and -E) and an 11S regulatory subcomplex (11SR) containing three non-SMC subunits (XCAP-D2, -G, and -H). We report here the biochemical and functional dissection of this chromosome condensation machinery. Although both 8SC and 13SC can bind to DNA in vitro and contain the SMC ATPase subunits, only 13SC is active as a DNA stimulated ATPase and supports ATP-dependent supercoiling activity. In the cell free extracts, 13SC is the active form that binds to chromosomes and induces their condensation. Neither 11SR nor 8SC alone is able to bind to chromatin. Our results suggest that the non-SMC subunits have dual roles in the regulation of condensin functions: one is to activate SMC ATPases and the other is to allow the holocomplex to associate with chromatin in a mitosis-specific manner. PMID- 11027309 TI - Discovering functional relationships between RNA expression and chemotherapeutic susceptibility using relevance networks. AB - In an effort to find gene regulatory networks and clusters of genes that affect cancer susceptibility to anticancer agents, we joined a database with baseline expression levels of 7,245 genes measured by using microarrays in 60 cancer cell lines, to a database with the amounts of 5,084 anticancer agents needed to inhibit growth of those same cell lines. Comprehensive pair-wise correlations were calculated between gene expression and measures of agent susceptibility. Associations weaker than a threshold strength were removed, leaving networks of highly correlated genes and agents called relevance networks. Hypotheses for potential single-gene determinants of anticancer agent susceptibility were constructed. The effect of random chance in the large number of calculations performed was empirically determined by repeated random permutation testing; only associations stronger than those seen in multiply permuted data were used in clustering. We discuss the advantages of this methodology over alternative approaches, such as phylogenetic-type tree clustering and self-organizing maps. PMID- 11027310 TI - The visual paired-comparison task as a measure of declarative memory. AB - Performance on the visual paired-comparison task depends on the integrity of the hippocampal formation in humans, monkeys, and, for an analogous task, in rats. The present study sought additional evidence in healthy volunteers concerning the nature of this task. We found that performance on the visual paired-comparison task was predictive of subsequent recognition memory performance whereas perceptual priming was unrelated to subsequent recognition memory performance. The results are consistent with the data from lesions and suggest that performance on the visual paired-comparison task measures a form of declarative memory. PMID- 11027311 TI - The Ebola virus VP35 protein functions as a type I IFN antagonist. AB - An assay has been developed that allows the identification of molecules that function as type I IFN antagonists. Using this assay, we have identified an Ebola virus-encoded inhibitor of the type I IFN response, the Ebola virus VP35 protein. The assay relies on the properties of an influenza virus mutant, influenza delNS1 virus, which lacks the NS1 ORF and, therefore, does not produce the NS1 protein. When cells are infected with influenza delNS1 virus, large amounts of type I IFN are produced. As a consequence, influenza delNS1 virus replicates poorly. However, high-efficiency transient transfection of a plasmid encoding a protein that interferes with type I IFN-induced antiviral functions, such as the influenza A virus NS1 protein or the herpes simplex virus protein ICP34.5, rescues growth of influenza delNS1 virus. When plasmids expressing individual Ebola virus proteins were transfected into Madin Darby canine kidney cells, the Ebola virus VP35 protein enhanced influenza delNS1 virus growth more than 100 fold. VP35 subsequently was shown to block double-stranded RNA- and virus mediated induction of an IFN-stimulated response element reporter gene and to block double-stranded RNA- and virus-mediated induction of the IFN-beta promoter. The Ebola virus VP35 therefore is likely to inhibit induction of type I IFN in Ebola virus-infected cells and may be an important determinant of Ebola virus virulence in vivo. PMID- 11027312 TI - A metabolic defect promotes obesity in mice lacking melanocortin-4 receptors. AB - Melanocortin-4 receptor (Mc4r)-null mice exhibit late-onset obesity. To determine whether aberrant metabolism contributes to the obesity, food consumption by Mc4r null mice was restricted to (pair-fed to) that consumed by wild-type (WT) mice. Pair-fed Mc4r-null females maintained body weights intermediate to that of WT and nonpair-fed Mc4r-null females, whereas pairfeeding normalized the body weights of Mc4r-null male mice. Fat pad and circulating leptin levels were elevated in both male and female pair-fed Mc4r-null mice compared with WT mice. Oxygen consumption of Mc4r-null mice with similar body weights as WT controls was reduced by 20%. Locomotor activity of young nonobese Mc4r-null males was significantly lower than that of WT males; however, locomotion of young nonobese females was normal. Core body temperature of Mc4r-null mice was normal, and they responded normally to cold exposure. Young nonobese Mc4r-null females were unable to induce uncoupling protein 1 (UCP1) in brown adipose tissue in response to peripheral leptin administration, whereas UCP1 mRNA was increased by 60% in the WT females. These results indicate that Mc4r deficiency enhances caloric efficiency, similar to that seen in the agouti obesity syndrome and in melanocortin-3 receptor-null mice. PMID- 11027313 TI - Cytoplasmic catalytic subunit of protein kinase A mediates cross-repression by NF kappa B and the glucocorticoid receptor. AB - Negative transcriptional regulation or cross-coupling between NF-kappa B (RelA) and the glucocorticoid receptor (GR) is proposed to play a regulatory role in human physiology and disease. Despite previous advances, the biochemical basis of this phenomenon remains a subject of controversy. We show here that the inhibition of GR activity by RelA does not require the RelA DNA binding, transactivation, or nuclear localization domains. Surprisingly, RelA repression of GR is abolished by mutation of the conserved protein kinase A (PKA) site at amino acid residue 276 of RelA. We show that GR associates in vivo and in vitro with the catalytic subunit of PKA (PKAc) in a ligand-independent manner and that GR transcription depends on PKA signaling. Indeed, we demonstrated that GR mediated inhibition of NF-kappa B transactivation is PKAc-dependent. In contrast to previous models, we suggest that the cross-coupling requires a cytoplasmic step and is regulated by a PKAc-associated signaling. PMID- 11027314 TI - Induction of primary NY-ESO-1 immunity: CD8+ T lymphocyte and antibody responses in peptide-vaccinated patients with NY-ESO-1+ cancers. AB - Cancer-testis antigen NY-ESO-1 is one of the most immunogenic tumor antigens defined to date. Spontaneous humoral and CD8+ T-cell responses to NY-ESO-1 are detected in 40-50% of patients with advanced NY-ESO-1-expressing tumors. A clinical trial was initiated to study the immunological effects of intradermal vaccination with 3 HLA-A2-binding NY-ESO-1 peptides in 12 patients with metastatic NY-ESO-1-expressing cancers. Seven patients were NY-ESO-1 serum antibody negative, and five patients were NY-ESO-1 serum antibody positive at the outset of the study. Primary peptide-specific CD8+ T-cell reactions and delayed type hypersensitivity responses were generated in four of seven NY-ESO-1 antibody negative patients. Induction of a specific CD8+ T-cell response to NY-ESO-1 in immunized antibody-negative patients was associated with disease stabilization and objective regression of single metastases. NY-ESO-1 antibody-positive patients did not develop significant changes in baseline NY-ESO-1-specific T-cell reactivity. However, stabilization of disease and regression of individual metastases were observed in three of five immunized patients. These results demonstrate that primary NY-ESO-1-specific CD8+ T-cell responses can be induced by intradermal immunization with NY-ESO-1 peptides, and that immunization with NY ESO-1 may have the potential to alter the natural course of NY-ESO-1-expressing tumors. PMID- 11027315 TI - DNA microarray analysis of gene expression in response to physiological and genetic changes that affect tryptophan metabolism in Escherichia coli. AB - We investigated the global changes in mRNA abundance in Escherichia coli elicited by various perturbations of tryptophan metabolism. To do so we printed DNA microarrays containing 95% of all annotated E. coli ORFs. We determined the expression profile that is predominantly dictated by the activity of the tryptophan repressor. Only three operons, trp, mtr, and aroH, exhibited appreciable expression changes consistent with this profile. The quantitative changes we observed in mRNA levels for the five genes of the trp operon were consistent within a factor of 2, with expectations based on established Trp protein levels. Several operons known to be regulated by the TyrR protein, aroF tyrA, aroL, aroP, and aroG, were down-regulated on addition of tryptophan. TyrR can be activated by any one of the three aromatic amino acids. Only one operon, tnaAB, was significantly activated by the presence of tryptophan in the medium. We uncovered a plethora of likely indirect effects of changes in tryptophan metabolism on intracellular mRNA pools, most prominent of which was the sensitivity of arginine biosynthetic operons to tryptophan starvation. PMID- 11027316 TI - Regulation of T cell activation, anxiety, and male aggression by RGS2. AB - Regulators of G protein signaling (RGS) proteins accelerate the GTPase activity of Galpha protein subunits in vitro, negatively regulating G protein-coupled receptor signaling. The physiological role of mammalian RGS proteins is largely unknown. The RGS family member rgs2 was cloned as an immediate early response gene up-regulated in T lymphocytes after activation. To investigate the role of RGS2 in vivo, we generated rgs2-deficient mice. We show that targeted mutation of rgs2 in mice leads to reduced T cell proliferation and IL-2 production, which translates in an impaired antiviral immunity in vivo. Interestingly, rgs2(-/-) mice also display increased anxiety responses and decreased male aggression in the absence of cognitive or motor deficits. RGS2 also controls synaptic development and basal electrical activity in hippocampal CA1 neurons. Thus, RGS2 plays an important role in T cell activation, synapse development in the hippocampus, and emotive behaviors. PMID- 11027317 TI - ABA activates multiple Ca(2+) fluxes in stomatal guard cells, triggering vacuolar K(+)(Rb(+)) release. AB - The mechanisms by which abscisic acid (ABA) activates the release of K(+)(Rb(+)) from the vacuole of stomatal guard cells, a process essential for ABA-induced stomatal closure, have been investigated by tracer flux measurements. The form and timing of the ABA-induced efflux transient could be manipulated by treatments that alter three potential Ca(2+) fluxes into the cytoplasm, the influx from the outside and two pathways of internal release, those dependent on phospholipase C (inhibited by ) and cyclic ADP-ribose (inhibited by nicotinamide). Ba(2+), acting as a competitive inhibitor of Ca(2+) influx but also as an inhibitor of internal release, was an effective inhibitor of the transient. The results suggest that a threshold level of cytoplasmic Ca(2+) is required for the initiation of the minimal efflux transient after a lag period and with a low rate of rise. As conditions improve for the generation of an efflux transient (higher ABA or reduced Ba(2+)), a second threshold is crossed, generating a transient with zero lag and rapid rate of rise. This may reflect different Ca(2+) levels required for activation of different tonoplast K(+) channels. In this state, at high ABA, the transient is inhibited by removal of external Ca(2+), suggesting Ca(2+) influx makes a major contribution to increase in cytoplasmic Ca(2+). By contrast, at low ABA, the transient is not inhibited by removal of external Ca(2+) but is sensitive to either or nicotinamide, suggesting internal release makes the major contribution, involving both pathways. ABA appears to activate all three processes, and their relative importance depends on conditions. PMID- 11027319 TI - Sticking together. PMID- 11027318 TI - A Saccharomyces gene family involved in invasive growth, cell-cell adhesion, and mating. AB - The cell wall of bakers' yeast contains a family of glycosyl-phosphatidylinositol (GPI)-linked glycoproteins of domain structure similar to the adhesins of pathogenic fungi. In wild-type cells each of these proteins has a unique function in different developmental processes (mating, invasive growth, cell-cell adhesion, or filamentation). What unifies these developmental events is adhesion, either to an inert substrate or to a cell. Although they differ in their specificities, many of these proteins can substitute for each other when overexpressed. For example, Flo11p is required during vegetative growth for haploid invasion and diploid filamentation, whereas Fig2p is required for mating. When overexpressed, Flo11p and Fig2p can function in mating, invasion, filamentation, and flocculation. The ability of Flo11p to supply Fig2p function in mating depends on its intracellular localization to the mating projection, where Fig2p normally functions in the adhesion of mating cells. Our data show that even distant family members retain the ability to carry out disparate functions if localized and expressed appropriately. PMID- 11027320 TI - Introduction to the Chinese-American Frontiers of Science. PMID- 11027321 TI - Assessing climate impacts. AB - Assessing climate impacts involves identifying sources and characteristics of climate variability, and mitigating potential negative impacts of that variability. Associated research focuses on climate driving mechanisms, biosphere hydrosphere responses and mediation, and human responses. Examples of climate impacts come from 1998 flooding in the Yangtze River Basin and hurricanes in the Caribbean and Central America. Although we have limited understanding of the fundamental driving-response interactions associated with climate variability, increasingly powerful measurement and modeling techniques make assessing climate impacts a rapidly developing frontier of science. PMID- 11027322 TI - Data assimilation and its applications. AB - In data assimilation, one prepares the grid data as the best possible estimate of the true initial state of a considered system by merging various measurements irregularly distributed in space and time, with a prior knowledge of the state given by a numerical model. Because it may improve forecasting or modeling and increase physical understanding of considered systems, data assimilation now plays a very important role in studies of atmospheric and oceanic problems. Here, three examples are presented to illustrate the use of new types of observations and the ability of improving forecasting or modeling. PMID- 11027323 TI - Macroscopic phase separation in high-temperature superconductors. AB - High-temperature superconductivity is recovered by introducing extra holes to the Cu-O planes, which initially are insulating with antiferromagnetism. In this paper I present data to show the macroscopic electronic phase separation that is caused by either mobile doping or electronic instability in the overdoped region. My results clearly demonstrate that the electronic inhomogeneity is probably a general feature of high-temperature superconductors. PMID- 11027324 TI - Novel polymers: molecular to nanoscale order in three dimensions. AB - The assembly of polymer chains in solution is a powerful method that is leading to the preparation of interesting and unique macromolecular-based synthetic nanostructures. Specific control over the intramolecular and intermolecular physical interactions dictates either the folding of single chains or the aggregation and ordering of multiple chains. This control is provided through the selective placement of functional groups along the polymer backbone and the relative strengths of their attractive and repulsive interactions. PMID- 11027325 TI - Nucleic acid-triggered catalytic drug release. AB - We propose a concept for the rational design and synthesis of highly selective chemotherapeutic agents that makes direct use of genetic information about the disease state. The key idea is to use the mRNA or DNA specific to the disease state to trigger the catalytic release of a cytotoxic drug by promoting the association of a prodrug with a catalyst capable of releasing the drug. We demonstrate the feasibility of such an approach in vitro with a model system that is based on the hydrolysis of p-nitrophenyl esters by imidazole. In our model system, the catalytic component consists of an imidazole group linked to the 5' end of a 15-mer that is complementary to the 5' end of the triggering oligodeoxynucleotide. The corresponding prodrug component consists of a p nitrophenol ester linked to the 3' end of an 8-mer oligodeoxynucleotide that is complementary to the 3' end of the triggering sequence. We show that this system efficiently releases p-nitrophenol in the presence of all three components and that the reaction is catalytic and undergoes multiple turnovers. We also show that the complex between the catalytic component and the triggering oligodeoxynucleotide behaves like an enzyme and follows Michaelis-Menten kinetics, with a K(M) of 22 microM and a k(cat) of 0.018 min(-1). Most importantly, we show that catalytic release of p-nitrophenol is sensitive to the presence of a single base-pair mismatch. PMID- 11027326 TI - Catalytic tempering: A method for sampling rough energy landscapes by Monte Carlo. AB - A new Monte Carlo algorithm is presented for the efficient sampling of the Boltzmann distribution of configurations of systems with rough energy landscapes. The method is based on the introduction of a fictitious coordinate y so that the dimensionality of the system is increased by one. This augmented system has a potential surface and a temperature that is made to depend on the new coordinate y in such a way that for a small strip of the y space, called the "normal region," the temperature is set equal to the temperature desired and the potential is the original rough energy potential. To enhance barrier crossing outside the "normal region," the energy barriers are reduced by truncation (with preservation of the potential minima) and the temperature is made to increase with ||y ||. The method, called catalytic tempering or CAT, is found to greatly improve the rate of convergence of Monte Carlo sampling in model systems and to eliminate the quasi-ergodic behavior often found in the sampling of rough energy landscapes. PMID- 11027327 TI - Single chain spectroscopy of conformational dependence of conjugated polymer photophysics. AB - Single molecule confocal fluorescence microscopy was used to perform photoluminescence spectroscopy on single, isolated molecules of the conjugated polymer poly[2-methoxy, 5-(2'-ethyl-hexyloxy)-p-phenylene-vinylene] (MEH-PPV). We show that the fluorescence from single chains of this electroluminescent polymer depends strongly on chain conformation. The time evolution of the spectra, emission intensity, and polarization all provide direct evidence that memory of the chain conformation in solution is retained after solvent evaporation. Chains cast from toluene solution are highly folded and show memory of the excitation polarization. Exciton funneling to highly aggregated low energy regions causes the chain to mimic the photophysical behavior of a single chromophore. Chains cast from chloroform, however, behave as multichromophore systems, and no sudden discrete spectral or intensity jumps are observed. These also exhibit different spectroscopy from the folded chromophores. PMID- 11027328 TI - Candidate gene analysis of organ pigmentation loci in the Solanaceae. AB - Ten structural genes from the Capsicum (pepper) carotenoid biosynthetic pathway have been localized on a (Capsicum annuum x Capsicum chinense)F(2) genetic map anchored in Lycopersicon (tomato). The positions of these genes were compared with positions of the same genes in tomato when known, and with loci from pepper, potato, and tomato that affect carotenoid levels in different tissues. C2, one of three phenotypically defined loci determining pepper fruit color, cosegregated with phytoene synthase. The capsanthin-capsorubin synthase (Ccs) locus, shown previously to cosegregate with Y, another pepper fruit color locus, mapped to pepper chromosome 6. Other structural genes in pepper corresponded to loci affecting carotenoid production as follows: Ccs in pepper and the B locus for hyperaccumulation of beta-carotene in tomato fruit mapped to homeologous regions; the position of the lycopene beta-cyclase gene in pepper may correspond to the lutescent-2 mutation in tomato; and the lycopene epsilon-cyclase locus in pepper corresponded to the lycopene epsilon-cyclase locus/Del mutation for hyperaccumulation of delta-carotene in tomato fruit. Additional associations were seen between the structural genes and previously mapped loci controlling quantitative variation in pepper and tomato fruit color. These results demonstrate that comparative analyses using candidate genes may be used to link specific metabolic phenotypes and loci that affect these phenotypes in related species. PMID- 11027329 TI - Distinct cAMP response element-binding protein (CREB) domains stimulate different steps in a concerted mechanism of transcription activation. AB - Hormones and neurotransmitters rapidly change patterns of gene expression in target cells by activating protein kinases that phosphorylate and modify the activity of CREB and other transcription factors. Although CREB was initially characterized as mediating the response to cAMP, CREB phosphorylation and activation are stimulated by diverse extracellular signals and protein kinases in essentially all cells and tissues. CREB stimulates transcription through a constitutive activation domain (CAD), which interacts with the promoter recognition factor TFIID, and through a kinase-inducible domain (KID), when Ser 133 is phosphorylated. The present study provides new insight into the mechanism of activation by showing that each of the CREB domains contributes to transcription initiation by stimulating sequential steps in the transcription reaction. The CAD effectively assembled a polymerase complex, as evidenced by constitutive activation in vivo and stimulation of single-round transcription in vitro. In contrast, phosphorylation of the KID in CREB stimulated isomerization of the polymerase complex, as determined by abortive initiation, and promoter clearance and/or reinitiation, as measured by multiple rounds of transcription. Our results provide evidence for a new model for CREB-mediated induction through a concerted mechanism involving establishment of a polymerase complex by the CAD, followed by stimulation of isomerization, promoter clearance, and/or reinitiation by phosphorylated KID to enhance target gene transcription. PMID- 11027330 TI - Intercellular delivery of a herpes simplex virus VP22 fusion protein from cells infected with lentiviral vectors. AB - Effective gene therapy depends on the efficient transfer of therapeutic genes and their protein products to target cells. Lentiviral vectors appear promising for virus-mediated gene delivery and long-term expression in nondividing cells. The herpes simplex virus type 1 tegument protein VP22 has recently been shown to mediate intercellular transport of proteins, raising the possibility that it may be helpful in a setting where the global delivery of therapeutic proteins is desired. To investigate the effectiveness of lentiviral vectors to deliver genes encoding proteins fused to VP22, and to test whether the system is sufficiently potent to allow protein delivery from transduced cells in vitro and in vivo, fusion constructs of VP22 and the enhanced green fluorescent protein (EGFP) were prepared and delivered into target cells by using HIV-1-based lentiviral vectors. To follow the spread of VP22-EGFP to other cells, transduced COS-7 cells were coplated with a number of different cell types, including brain choroid plexus cells, human endothelial cells, H9 cells, and HeLa cells. We found that VP22-EGFP fusion proteins were transported from transduced cells to recipient cells and that such fusion proteins accumulated in the nucleus and in the cytoplasm of such cells. To determine the ability to deliver fusion proteins in vivo, we injected transduced H9 cells as well as the viral vector directly into the brain of mice. We present evidence that VP22-EGFP fusion proteins were transported effectively from lentivirus transduced cells in vivo. We also show that the VP22-EGFP fusion protein encoded by the lentivirus is transported between cells. Our data indicate that such fusion proteins are present in the nucleus and in the cytoplasm of neighboring cells. Therefore, lentiviral vectors may provide a potent biological system for delivering genes encoding therapeutic proteins fused to VP22. PMID- 11027331 TI - Distinct but overlapping roles of histone acetylase PCAF and of the closely related PCAF-B/GCN5 in mouse embryogenesis. AB - PCAF plays a role in transcriptional activation, cell-cycle arrest, and cell differentiation in cultured cells. PCAF contributes to transcriptional activation by acetylating chromatin and transcription factors through its intrinsic histone acetylase activity. In this report, we present evidence for the in vivo function of PCAF and the closely related PCAF-B/GCN5. Mice lacking PCAF are developmentally normal without a distinct phenotype. In PCAF null-zygous mice, protein levels of PCAF-B/GCN5 are drastically elevated in lung and liver, where PCAF is abundantly expressed in wild-type mice, suggesting that PCAF-B/GCN5 functionally compensates for PCAF. In contrast, animals lacking PCAF-B/GCN5 die between days 9.5 and 11.5 of gestation. Normally, PCAF-B/GCN5 mRNA is expressed at high levels already by day 8, whereas PCAF mRNA is first detected on day 12.5, which may explain, in part, the distinct knockout phenotypes. These results provide evidence that PCAF and PCAF-B/GCN5 play distinct but functionally overlapping roles in embryogenesis. PMID- 11027332 TI - Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. AB - Human embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of in vitro fertilized human blastocysts. We examined the potential of eight growth factors [basic fibroblast growth factor (bFGF), transforming growth factor beta1 (TGF-beta1), activin-A, bone morphogenic protein 4 (BMP-4), hepatocyte growth factor (HGF), epidermal growth factor (EGF), beta nerve growth factor (betaNGF), and retinoic acid] to direct the differentiation of human ES-derived cells in vitro. We show that human ES cells that have initiated development as aggregates (embryoid bodies) express a receptor for each of these factors, and that their effects are evident by differentiation into cells with different epithelial or mesenchymal morphologies. Differentiation of the cells was assayed by expression of 24 cell-specific molecular markers that cover all embryonic germ layers and 11 different tissues. Each growth factor has a unique effect that may result from directed differentiation and/or cell selection, and we can divide the overall effects of the factors into three categories: growth factors (Activin-A and TGFbeta1) that mainly induce mesodermal cells; factors (retinoic acid, EGF, BMP-4, and bFGF) that activate ectodermal and mesodermal markers; and factors (NGF and HGF) that allow differentiation into the three embryonic germ layers, including endoderm. None of the growth factors directs differentiation exclusively to one cell type. This analysis sets the stage for directing differentiation of human ES cells in culture and indicates that multiple human cell types may be enriched in vitro by specific factors. PMID- 11027333 TI - Independent adaptation to riverine habitats allowed survival of ancient cetacean lineages. AB - The four species of "river dolphins" are associated with six separate great river systems on three subcontinents and have been grouped for more than a century into a single taxon based on their similar appearance. However, several morphologists recently questioned the monophyly of that group. By using phylogenetic analyses of nucleotide sequences from three mitochondrial and two nuclear genes, we demonstrate with statistical significance that extant river dolphins are not monophyletic and suggest that they are relict species whose adaptation to riverine habitats incidentally insured their survival against major environmental changes in the marine ecosystem or the emergence of Delphinidae. PMID- 11027334 TI - Heat shock protein 60 sequence comparisons: duplications, lateral transfer, and mitochondrial evolution. AB - Heat shock proteins 60 (GroEL) are highly expressed essential proteins in eubacterial genomes and in eukaryotic organelles. These chaperone proteins have been advanced as propitious marker sequences for tracing the evolution of mitochondrial (Mt) genomes. Similarities among HSP60 sequences based on significant segment pair alignment calculations are used to deduce associations of sequences taking into account GroEL functional/structural domain differences and to relate HSP60 duplications pervasive in alpha-proteobacterial lineages to the dynamics of lateral transfer and plasmid integration. Multiple alignments with consensuses are determined for 10 natural groups. The group consensuses sharpen the similarity contrasts among individual sequences. In particular, the Mt group matches best with the classical alpha-proteobacteria and closely with Rickettsia but significantly worse with the rickettsial groups Ehrlichia and Orientia. However, across broad protein sequence comparisons, there appears to be no consistent prokaryote whose protein sequences align best with animal Mt genomes. There are plausible scenarios indicating that the nuclear-encoded HSP60 (and HSP70) sequences functioning in Mt are results of lateral transfer and are probably derived from an alpha-proteobacterium. This hypothesis relates to the plethora of duplicated HSP60 sequences among the classical alpha-proteobacteria contrasted with no duplications of HSP60 among other clades of proteobacterial genomes. Evolutionary relations are confounded by differential selection pressures, convergence, variable mutational rates, site variability, and lateral gene transfer. PMID- 11027335 TI - Changes in mitochondrial genetic codes as phylogenetic characters: two examples from the flatworms. AB - Shared molecular genetic characteristics other than DNA and protein sequences can provide excellent sources of phylogenetic information, particularly if they are complex and rare and are consequently unlikely to have arisen by chance convergence. We have used two such characters, arising from changes in mitochondrial genetic code, to define a clade within the Platyhelminthes (flatworms), the Rhabditophora. We have sampled 10 distinct classes within the Rhabditophora and find that all have the codon AAA coding for the amino acid Asn rather than the usual Lys and AUA for Ile rather than the usual Met. We find no evidence to support claims that the codon UAA codes for Tyr in the Platyhelminthes rather than the standard stop codon. The Rhabditophora are a very diverse group comprising the majority of the free-living turbellarian taxa and the parasitic Neodermata. In contrast, three other classes of turbellarian flatworm, the Acoela, Nemertodermatida, and Catenulida, have the standard invertebrate assignments for these codons and so are convincingly excluded from the rhabditophoran clade. We have developed a rapid computerized method for analyzing genetic codes and demonstrate the wide phylogenetic distribution of the standard invertebrate code as well as confirming already known metazoan deviations from it (ascidian, vertebrate, echinoderm/hemichordate). PMID- 11027336 TI - Werner protein recruits DNA polymerase delta to the nucleolus. AB - Werner syndrome is a Mendelian disorder of man that produces a number of manifestations resembling human aging. This disorder is caused by inactivation of the wrn gene, a member of the RecQ family of DNA helicases. The helicase and exonuclease activities of the Werner protein (WRN) suggest that it functions in DNA transactions, but the physiological function of WRN remains elusive. We present several lines of evidence that WRN interacts specifically with the p50 subunit of polymerase delta, the major DNA polymerase required for chromosomal DNA replication. P50, identified by yeast two-hybrid screening, interacts physically with the C terminus of WRN. Native WRN protein coimmunoprecipitates with p50 in a cellular fraction enriched in nucleolar proteins, and this immunocomplex also includes p125, the catalytic subunit of polymerase delta. In subcellular localization studies of cells transfected with WRN, p50 and p125 redistribute to the nucleolus and colocalize with WRN. These results suggest that one of the functions of WRN protein is to directly modify DNA replication via its interaction with p50 and abet dynamic relocalization of the DNA polymerase delta complexes within the nucleus. PMID- 11027337 TI - The expression of adipogenic genes is decreased in obesity and diabetes mellitus. AB - Obesity is strongly correlated with type 2 diabetes mellitus, a common disorder of glucose and lipid metabolism. Although adipocytes are critical in obesity, their role in diabetes has only recently been appreciated. We conducted studies by using DNA microarrays to identify differences in gene expression in adipose tissue from lean, obese, and obese-diabetic mice. The expression level of over 11,000 transcripts was analyzed, and 214 transcripts showed significant differences between lean and obese mice. Surprisingly, the expression of genes normally associated with adipocyte differentiation were down-regulated in obesity. Not all obese individuals will become diabetic; many remain normoglycemic despite profound obesity. Understanding the transition to obesity with concomitant diabetes will provide important clues to the pathogenesis of type 2 diabetes. Therefore, we examined the levels of gene expression in adipose tissue from five groups of obese mice with varying degrees of hyperglycemia, and we identified 88 genes whose expression strongly correlated with diabetes severity. This group included many genes that are known to be involved in signal transduction and energy metabolism as well as genes not previously examined in the context of diabetes. Our data show that a decrease in expression of genes normally involved in adipogenesis is associated with obesity, and we further identify genes important for subsequent development of type 2 diabetes mellitus. PMID- 11027338 TI - A Ufd2/D4Cole1e chimeric protein and overexpression of Rbp7 in the slow Wallerian degeneration (WldS) mouse. AB - Exons of three genes were identified within the 85-kilobase tandem triplication unit of the slow Wallerian degeneration mutant mouse, C57BL/Wld(S). Ubiquitin fusion degradation protein 2 (Ufd2) and a previously undescribed gene, D4Cole1e, span the proximal and distal boundaries of the repeat unit, respectively. They have the same chromosomal orientation and form a chimeric gene when brought together at the boundaries between adjacent repeat units in Wld(S). The chimeric mRNA is abundantly expressed in the nervous system and encodes an in-frame fusion protein consisting of the N-terminal 70 amino acids of Ufd2, the C-terminal 302 amino acids of D4Cole1e, and an aspartic acid formed at the junction. Antisera raised against synthetic peptides detect the expected 43-kDa protein specifically in Wld(S) brain. This expression pattern, together with the previously established role of ubiquitination in axon degeneration, makes the chimeric gene a promising candidate for Wld. The third gene altered by the triplication, Rbp7, is a novel member of the cellular retinoid-binding protein family and is highly expressed in white adipose tissue and mammary gland. The whole gene lies within the repeat unit leading to overexpression of the normal transcript in Wld(S) mice. However, it is undetectable on Northern blots of Wld(S) brain and seems unlikely to be the Wld gene. These data reveal both a candidate gene for Wld and the potential of the Wld(S) mutant for studies of ubiquitin and retinoid metabolism. PMID- 11027339 TI - Global mapping of meiotic recombination hotspots and coldspots in the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, meiotic recombination is initiated by double-strand DNA breaks (DSBs). Meiotic DSBs occur at relatively high frequencies in some genomic regions (hotspots) and relatively low frequencies in others (coldspots). We used DNA microarrays to estimate variation in the level of nearby meiotic DSBs for all 6,200 yeast genes. Hotspots were nonrandomly associated with regions of high G + C base composition and certain transcriptional profiles. Coldspots were nonrandomly associated with the centromeres and telomeres. PMID- 11027340 TI - Identification of a functional transposase of the Tol2 element, an Ac-like element from the Japanese medaka fish, and its transposition in the zebrafish germ lineage. AB - The Tol2 element of the medaka fish Oryzias latipes belongs to the hAT family of transposons (hobo/Ac/Tam3). We report here identification of a functional transposase of Tol2 that is capable of catalyzing its transposition in the germ line of zebrafish Danio rerio. A transcript produced from Tol2 encodes a putative transposase. Zebrafish fertilized eggs were coinjected with mRNA transcribed in vitro, using cDNA of the Tol2 transcript as a template and a plasmid DNA harboring a mutant Tol2, which had a deletion in the putative transposase gene but retained necessary cis sequences. The injected fish were raised to adulthood and mated to noninjected fish, and genomic DNA of the progeny fish were analyzed by PCR and Southern hybridization. Half of F(1) fish obtained from one of eight injected fish contained the Tol2 DNA in their genomes but not the vector portion. Among these F(1) fish, Tol2 insertions at four different loci were identified, and some F(1) fish carried two or three different Tol2 insertions, indicating that the germ line of the founder fish is highly mosaic. Sequencing analyses revealed that, in all cases, Tol2 was surrounded by zebrafish genomic sequences, and an 8-bp duplication was created at the target site, indicating that Tol2 was integrated in the zebrafish genome through transposition. This study identifies an autonomous member of a DNA-based transposable element from a vertebrate genome. The Tol2 transposon system should thus be used to develop novel transgenesis and insertional mutagenesis methods in zebrafish and possibly in other fishes. PMID- 11027341 TI - Evolution of the recombination signal sequences in the Ig heavy-chain variable region locus of mammals. AB - The Ig and T cell receptor (TCR) loci have an exceptionally dynamic evolutionary history, but the mechanisms responsible remain a subject of speculation. Ig and TCR genes are unique in vertebrates in that they are assembled from V, D, and J segments by site-specific recombination in developing lymphocytes. Here we examine the extent to which the V(D)J recombination in germline cells may have been responsible for remodeling Ig and TCR loci in mammals by asking whether gene segments have evolved as a unit, or whether, instead, recombination signal sequences (RSSs) and coding sequences have different phylogenies. Four distinct types of RSS have been defined in the human Ig heavy-chain variable region (Vh) locus, namely H1, H2, H3, and H5, and no other RSS type has been detected in other mammalian species. There is a well-supported discrepancy between the evolutionary history of the RSSs as compared with the Vh coding sequences: the RSS type H2 of one Vh gene segment has clearly become replaced by a RSS type H3 during mammalian evolution, between 115 and 65 million years ago. Two general models might explain the RSS swap: the first involves an unequal crossing over, and the second implicates germline activation of V(D)J recombination. The Vh H2/RSS-H3 recombination product has likely been selected during the evolution of mammals because it provides better V(D)J recombination efficiency. PMID- 11027342 TI - RelB nuclear translocation regulates B cell MHC molecule, CD40 expression, and antigen-presenting cell function. AB - Mice with targeted RelB mutations demonstrated an essential role for RelB in immune responses and in myeloid dendritic cell differentiation. Human studies suggested a more global transcriptional role in antigen presentation. Burkitt lymphoma cell lines were used as a model to examine the role of RelB in antigen presentation. After transient transfection of BJAB with RelB, strong nuclear expression of RelB-p50 heterodimers was associated with increased APC function and expression of CD40 and MHC class I. Antisense RelB in DG75 reduced antigen presenting capacity and CD40-mediated up-regulation of MHC molecules. The data indicate that RelB transcriptional activity directly affects antigen presentation and CD40 synthesis. Stimulation of RelB transcriptional activity may provide a positive feedback loop for facilitating productive APC/T cell interactions. PMID- 11027344 TI - Genetic susceptibility to breast cancer: HLA DQB*03032 and HLA DRB1*11 may represent protective alleles. AB - Tumors are believed to emerge only when immune surveillance fails. We wished to ascertain whether the failure to inherit putative protective alleles of HLA class II genes is linked to the development of breast cancer. We molecularly typed HLA DPB1, DQB1, DRB1, and DRB3 alleles in 176 Caucasian women diagnosed with early onset breast cancer and in 215 ethnically matched controls. HLA DQB*03032 was identified in 7% of controls but in no patients with early-onset breast cancer (P = 0.0001). HLA DRB1*11 alleles were also significantly overrepresented (P < 0.0001) in controls (16. 3%) as compared with patients with early-onset breast cancer (3.5%). HLA DQB*03032 and HLA DRB1*11 alleles may have a protective role in human breast cancer. PMID- 11027343 TI - Constitutive expression of a complement-like protein in toll and JAK gain-of function mutants of Drosophila. AB - We show that Drosophila expresses four genes encoding proteins with significant similarities with the thiolester-containing proteins of the complement C3/alpha(2)-macroglobulin superfamily. The genes are transcribed at a low level during all stages of development, and their expression is markedly up-regulated after an immune challenge. For one of these genes, which is predominantly expressed in the larval fat body, we observe a constitutive expression in gain-of function mutants of the Janus kinase (JAK) hop and a reduced inducibility in loss of-function hop mutants. We also observe a constitutive expression in gain-of function Toll mutants. We discuss the possible roles of these novel complement like proteins in the Drosophila host defense. PMID- 11027345 TI - Immunization against Alzheimer's beta -amyloid plaques via EFRH phage administration. AB - The epitope EFRH, corresponding to amino acids 3-6 within the human beta-amyloid peptide (AbetaP), acts as a regulatory site controlling both the formation and disaggregation process of the beta-amyloid fibrils (Abeta). Locking of this epitope by highly specific antibodies affects the dynamics of the entire AbetaP molecule, preventing self-aggregation as well as enabling resolubilization of already formed aggregates. Production of such antibodies by repeated injections of toxic human Abeta fibrils into transgenic mice suggests the feasibility of vaccination against Alzheimer's disease. Here, we report the development of an immunization procedure for the production of effective anti-aggregating beta amyloid antibodies based on filamentous phages displaying the EFRH peptide as specific and nontoxic antigen. Effective autoimmune antibodies were obtained by EFRH phage administration in guinea pigs, which exhibit AbetaP identical to the human AbetaP region. Moreover, because of the high antigenicity of the phage, no adjuvant is required to obtain high affinity anti-aggregating IgG antibodies after a short immunization period of 3 weeks. Availability of such antibodies opens up possibilities for the development of an efficient and long-lasting vaccination for the prevention and treatment of Alzheimer's disease. PMID- 11027346 TI - Selective CXCR4 antagonism by Tat: implications for in vivo expansion of coreceptor use by HIV-1. AB - Chemokines and chemokine receptors play important roles in HIV-1 infection and tropism. CCR5 is the major macrophage-tropic coreceptor for HIV-1 whereas CXC chemokine receptor 4 (CXCR4) serves the counterpart function for T cell-tropic viruses. An outstanding biological mystery is why only R5-HIV-1 is initially detected in new seroconvertors who are exposed to R5 and X4 viruses. Indeed, X4 virus emerges in a minority of patients and only in the late stage of disease, suggesting that early negative selection against HIV-1-CXCR4 interaction may exist. Here, we report that the HIV-1 Tat protein, which is secreted from virus infected cells, is a CXCR4-specific antagonist. Soluble Tat selectively inhibited the entry and replication of X4, but not R5, virus in peripheral blood mononuclear cells (PBMCs). We propose that one functional consequence of secreted Tat is to select against X4 viruses, thereby influencing the early in vivo course of HIV-1 disease. PMID- 11027347 TI - Specific Th2 cells accumulate in the central nervous system of mice protected against experimental autoimmune encephalomyelitis by copolymer 1. AB - This study addresses the issue of the effect of immunomodulating therapies in the target organ-the central nervous system (CNS)-in the case of multiple sclerosis. Copolymer 1 (Cop 1, Copaxone, glatiramer acetate), an approved drug for the treatment of multiple sclerosis, is a potent inducer of Th2 regulatory cells in both mice and humans. Highly reactive Cop 1-specific T cell lines that secrete IL 4, IL-5, IL-6, IL-10, and transforming growth factor-beta in response to Cop 1 and crossreact with myelin basic protein (MBP) at the level of Th2 cytokine secretion were established from both brains and spinal cords of Cop 1-treated mice. In contrast, no reactivity to the control antigen lysozyme could be obtained in lymphocytes isolated from CNS of mice injected with lysozyme. Adoptively transferred labeled Cop 1-specific suppressor cells were found in brain sections 7 and 10 days after their injection to the periphery, whereas lysozyme-specific cells were absent in the CNS. Hence, Cop 1-induced Th2 cells cross the blood-brain barrier and accumulate in the CNS, where they can be stimulated in situ by MBP and thereby exert therapeutic effects in the diseased organ. This therapeutic effect was manifested, in brains of experimental autoimmune encephalomyelitis-induced mice, by a decrease in the inflammatory cytokine interferon-gamma and by secretion of the anti-inflammatory cytokine IL 10 in response to the autoantigen MBP. PMID- 11027348 TI - Detection of emphysema in rat lungs by using magnetic resonance measurements of 3He diffusion. AB - Emphysema is a pulmonary disease characterized by alveolar wall destruction, resulting in enlargement of gas exchange spaces without fibrosis. This condition is a part of chronic obstructive pulmonary disease (COPD), which causes 3.5% of deaths worldwide [Anonymous (1990) World Health Stat. Q. Special, 1-51] and contributes greatly to the global burden of disease [Murray, C. J. & Lopez, A. D. (1996) Science 274, 740-743]. Alveolar regeneration has been shown in animal models and could have potential for clinical treatment of early-stage emphysema. However, current techniques for detection of emphysema are not sensitive at the initial stages. Early-stage human panacinar emphysema is modeled in elastase treated animals. Here, we provide an in vivo imaging method for differentiating normal and emphysematous rat lungs by measuring the apparent diffusion coefficient (ADC) of hyperpolarized (3)He by using magnetic resonance imaging. These data show that the ADC is significantly larger in elastase-treated rats, indicating alveolar expansion. Whereas these rats were clinically asymptomatic, conventional histology confirmed presence of injury. Our results indicate that measurement of the hyperpolarized (3)He ADC can be a valuable research tool and has potential application in the clinical setting. PMID- 11027349 TI - Role of circulating nitrite and S-nitrosohemoglobin in the regulation of regional blood flow in humans. AB - To determine the relative contributions of endothelial-derived nitric oxide (NO) vs. intravascular nitrogen oxide species in the regulation of human blood flow, we simultaneously measured forearm blood flow and arterial and venous levels of plasma nitrite, LMW-SNOs and HMW-SNOs, and red cell S-nitrosohemoglobin (SNO-Hb). Measurements were made at rest and during regional inhibition of NO synthesis, followed by forearm exercise. Surprisingly, we found significant circulating arterial-venous plasma nitrite gradients, providing a novel delivery source for intravascular NO. Further supporting the notion that circulating nitrite is bioactive, the consumption of nitrite increased significantly with exercise during the inhibition of regional endothelial synthesis of NO. The role of circulating S-nitrosothiols and SNO-Hb in the regulation of basal vascular tone is less certain. We found that low-molecular-weight S-nitrosothiols were undetectable and S-nitroso-albumin levels were two logs lower than previously reported. In fact, S-nitroso-albumin primarily formed in the venous circulation, even during NO synthase inhibition. Whereas SNO-Hb was measurable in the human circulation (brachial artery levels of 170 nM in whole blood), arterial-venous gradients were not significant, and delivery of NO from SNO-Hb was minimal. In conclusion, we present data that suggest (i) circulating nitrite is bioactive and provides a delivery gradient of intravascular NO, (ii) S-nitroso-albumin does not deliver NO from the lungs to the tissue but forms in the peripheral circulation, and (iii) SNO-Hb and S-nitrosothiols play a minimal role in the regulation of basal vascular tone, even during exercise stress. PMID- 11027350 TI - An alternative interpretation of nanobacteria-induced biomineralization. AB - The reported isolation of nanobacteria from human kidney stones raises the intriguing possibility that these microorganisms are etiological agents of pathological extraskeletal calcification [Kajander, E. O. & Ciftcioglu, N. (1998) Proc. Natl. Acad. Sci. USA 95, 8274-8279]. Nanobacteria were previously isolated from FBS after prolonged incubation in DMEM. These bacteria initiated biomineralization of the culture medium and were identified in calcified particles and biofilms by nucleic acid stains, 16S rDNA sequencing, electron microscopy, and the demonstration of a transferable biomineralization activity. We have now identified putative nanobacteria, not only from FBS, but also from human saliva and dental plaque after the incubation of 0.45-microm membrane filtered samples in DMEM. Although biomineralization in our "cultures" was transferable to fresh DMEM, molecular examination of decalcified biofilms failed to detect nucleic acid or protein that would be expected from growth of a living entity. In addition, biomineralization was not inhibited by sodium azide. Furthermore, the 16S rDNA sequences previously ascribed to Nanobacterium sanguineum and Nanobacterium sp. were found to be indistinguishable from those of an environmental microorganism, Phyllobacterium mysinacearum, that has been previously detected as a contaminant in PCR. Thus, these data do not provide plausible support for the existence of a previously undiscovered bacterial genus. Instead, we provide evidence that biomineralization previously attributed to nanobacteria may be initiated by nonliving macromolecules and transferred on "subculture" by self-propagating microcrystalline apatite. PMID- 11027351 TI - Invasion in vitro of mosquito midgut cells by the malaria parasite proceeds by a conserved mechanism and results in death of the invaded midgut cells. AB - Using an in vitro culture system, we observed the migration of malaria ookinetes on the surface of the mosquito midgut and invasion of the midgut epithelium. Ookinetes display constrictions during migration to the midgut surface and a gliding motion once on the luminal midgut surface. Invasion of a midgut cell always occurs at its lateral apical surface. Invasion is rapid and is often followed by invasion of a neighboring midgut cell by the ookinete. The morphology of the invaded cells changes dramatically after invasion, and invaded cells die rapidly. Midgut cell death is accompanied by activation of a caspase-3-like protease, suggesting cell death is apoptotic. The events occurring during invasion were identical for two different species of Plasmodium and two different genera of mosquitoes; they probably represent a universal mechanism of mosquito midgut penetration by the malaria parasite. PMID- 11027352 TI - A novel pH2 control on the expression of flagella in the hyperthermophilic strictly hydrogenotrophic methanarchaeaon Methanococcus jannaschii. AB - The methanarchaeon, Methanococcus jannaschii, a hyperthermophilic, autotrophic, and strictly hydrogenotrophic inhabitant of submarine hydrothermal vents, was cultivated in a reactor at two hydrogen partial pressure (p(H(2))) values, 178 kPa (high) and 650 Pa (ultralow), and the cells were subjected to a comparative proteome analysis. From these studies, it was discovered that, when p(H(2)) was high and the cell density was low (a combination representing a hydrogen-excess condition), the cells possessed very low or undetectable levels of four flagella related polypeptides (FlaB2, FlaB3, FlaD, and FlaE); electron microscopic examination showed that most of these cells were devoid of flagella. Flagella synthesis occurred when hydrogen became limiting either at high cell density under high p(H(2)) or at low cell density under low p(H(2)). The results from a p(H(2))-shift experiment corroborated the above observations. The p(H(2)) dependent changes in the levels of two methanogenic enzymes (MTD and HMDX) were as expected, and thus they served as internal controls. To our knowledge, this is the first example for the regulation of expression of flagella by hydrogen in any domain of life and for a control of any kind on flagella synthesis in the archaea. Our work also provides the only known example for each of the following: (i) the pure culture cultivation of a methanogen at an ultralow, near ecologically relevant p(H(2)); (ii) experimental functional genomics for M. jannaschii; and (iii) the use of proteomics with M. jannaschii. PMID- 11027353 TI - Stimulation of glutamate receptor protein synthesis and membrane insertion within isolated neuronal dendrites. AB - The selective subcellular localization of mRNAs to dendrites and the recent demonstration of local protein synthesis have highlighted the potential role of postsynaptic sites in modulation of cell-cell communication. We show that epitope tagged subunit 2 of the ionotopic glutamate receptor, GluR2, mRNA transfected into isolated hippocampal neuronal dendrites is translated in response to pharmacologic stimulation. Further, confocal imaging of N-terminally labeled GluR2 reveals that the newly synthesized GluR2 protein can integrate into the dendritic membrane with the N terminus externally localized. These data demonstrate that integral membrane proteins can be synthesized in dendrites and can locally integrate into the cell membrane. PMID- 11027354 TI - Overexpression of the human VPAC2 receptor in the suprachiasmatic nucleus alters the circadian phenotype of mice. AB - The neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) belong to a superfamily of structurally related peptide hormones that includes glucagon, glucagon-like peptides, secretin, and growth hormone-releasing hormone. Microinjection of VIP or PACAP into the rodent suprachiasmatic nucleus (SCN) phase shifts the circadian pacemaker and VIP antagonists, and antisense oligodeoxynucleotides have been shown to disrupt circadian function. VIP and PACAP have equal potency as agonists of the VPAC(2) receptor (VPAC(2)R), which is expressed abundantly in the SCN, in a circadian manner. To determine whether manipulating the level of expression of the VPAC(2)R can influence the control of the circadian clock, we have created transgenic mice overexpressing the human VPAC(2)R gene from a yeast artificial chromosome (YAC) construct. The YAC was modified by a strategy using homologous recombination to introduce (i) the HA epitope tag sequence (from influenza virus hemagglutinin) at the carboxyl terminus of the VPAC(2)R protein, (ii) the lacZ reporter gene, and (iii) a conditional centromere, enabling YAC DNA to be amplified in culture in the presence of galactose. High levels of lacZ expression were detected in the SCN, habenula, pancreas, and testis of the transgenic mice, with lower levels in the olfactory bulb and various hypothalamic areas. Transgenic mice resynchronized more quickly than wild-type controls to an advance of 8 h in the light-dark (LD) cycle and exhibited a significantly shorter circadian period in constant darkness (DD). These data suggest that the VPAC(2)R can influence the rhythmicity and photic entrainment of the circadian clock. PMID- 11027355 TI - A novel function for serotonin-mediated short-term facilitation in aplysia: conversion of a transient, cell-wide homosynaptic hebbian plasticity into a persistent, protein synthesis-independent synapse-specific enhancement. AB - Studies of sensitization and classical conditioning of the gill-withdrawal reflex in Aplysia have shown that the synaptic connections between identified glutamatergic sensory neurons and motor neurons can be enhanced in one of two ways: by a heterosynaptic (modulatory input-dependent) mechanism that gives rise with repetition to long-term facilitation and by a homosynaptic (activity dependent) mechanism that gives rise with repetition to a facilitation that is partially blocked by 2-amino-5-phosphonovaleric acid and by injection of 1,2 bis(2-aminophenoxy)ethane-N,N,N', N'-tetraacetate (BAPTA) into the postsynaptic cell and is similar to long-term potentiation in the hippocampus. We here have examined how these two forms of facilitation interact at the level of an individual synaptic connection by using a culture preparation consisting of a single bifurcated sensory neuron that forms independent synaptic contacts with each of two spatially separated motor neurons. We find that the homosynaptic facilitation produced by a train of action potentials is cell wide and is evident at all of the terminals of the sensory neuron. By contrast, the heterosynaptic facilitation mediated by the modulatory transmitter serotonin (5-HT) can operate at the level of a single synapse. Homosynaptic activation gives rise to only a transient facilitation lasting a few hours, even when repeated in a spaced manner. The heterosynaptic facilitation produced by a single pulse of 5-HT, applied to one terminal of the sensory neuron, also lasts only minutes. However, when one or more homosynaptic trains of spike activity are paired with even a single pulse of 5-HT applied to one of the two branches of the sensory neuron, the combined actions lead to a selective enhancement in synaptic strength only at the 5-HT-treated branch that now lasts more than a day, and thus amplifies, by more than 20-fold, the duration of the individually produced homo- and heterosynaptic facilitation. This form of synapse-specific facilitation has unusual long-term properties. It does not require protein synthesis, nor is it accompanied by synaptic growth. PMID- 11027356 TI - Role of rab GDP dissociation inhibitor alpha in regulating plasticity of hippocampal neurotransmission. AB - Rab GDP dissociation inhibitor alpha (Rab GDIalpha) is a regulator of the Rab small G proteins implicated in neurotransmission, and mutations of Rab GDIalpha cause human X-linked mental retardation associated with epileptic seizures. In Rab GDIalpha-deficient mice, synaptic potentials in the CA1 region of the hippocampus displayed larger enhancement during repetitive stimulation, which was apparently opposite to the phenotype of Rab3A-deficient mice. Furthermore, the Rab GDIalpha-deficient mice showed hypersensitivity to bicuculline, an inducer of epileptic seizures. These results suggest that Rab GDIalpha plays a specialized role in Rab3A recycling to suppress hyperexcitability via modulation of presynaptic forms of plasticity. PMID- 11027357 TI - Sensory neuron-specific sodium channel SNS is abnormally expressed in the brains of mice with experimental allergic encephalomyelitis and humans with multiple sclerosis. AB - Clinical abnormalities in multiple sclerosis (MS) have classically been considered to be caused by demyelination and/or axonal degeneration; the possibility of molecular changes in neurons, such as the deployment of abnormal repertoires of ion channels that would alter neuronal electrogenic properties, has not been considered. Sensory Neuron-Specific sodium channel SNS displays a depolarized voltage dependence, slower activation and inactivation kinetics, and more rapid recovery from inactivation than classical "fast" sodium channels. SNS is selectively expressed in spinal sensory and trigeminal ganglion neurons within the peripheral nervous system and is not expressed within the normal brain. Here we show that sodium channel SNS mRNA and protein, which are not present within the cerebellum of control mice, are expressed within cerebellar Purkinje cells in a mouse model of MS, chronic relapsing experimental allergic encephalomyelitis. We also demonstrate SNS mRNA and protein expression within Purkinje cells from tissue obtained postmortem from patients with MS, but not in control subjects with no neurological disease. These results demonstrate a change in sodium channel expression in neurons within the brain in an animal model of MS and in humans with MS and suggest that abnormal patterns of neuronal ion channel expression may contribute to clinical abnormalities such as ataxia in these disorders. PMID- 11027358 TI - Nongenomic actions of estrogens and xenoestrogens by binding at a plasma membrane receptor unrelated to estrogen receptor alpha and estrogen receptor beta. AB - The molecular mechanism used by environmental chemicals to exert their hormone like actions is still only partially resolved. Although it generally is accepted that xenoestrogens act at the genomic level by binding to intracellular estrogen receptors, we have shown here that they trigger nongenomic effects in pancreatic beta cells. Both xenoestrogens and the circulating hormone, 17beta-estradiol, bind with high affinity to a common membrane binding site unrelated to the intracellular estrogen receptors ERalpha and ERbeta. This binding site is shared by dopamine, epinephrine, and norepinephrine and has the pharmacological profile of the gammaadrenergic receptor. This study provides an outline of the membrane receptor involved in rapid xenoestrogen actions. PMID- 11027359 TI - Nitric oxide prevents cardiovascular disease and determines survival in polyglobulic mice overexpressing erythropoietin. AB - Nitric oxide (NO) induces vasodilatatory, antiaggregatory, and antiproliferative effects in vitro. To delineate potential beneficial effects of NO in preventing vascular disease in vivo, we generated transgenic mice overexpressing human erythropoietin. These animals induce polyglobulia known to be associated with a high incidence of vascular disease. Despite hematocrit levels of 80%, adult transgenic mice did not develop hypertension or thromboembolism. Endothelial NO synthase levels, NO-mediated endothelium-dependent relaxation and circulating and vascular tissue NO levels were markedly increased. Administration of the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) led to vasoconstriction of peripheral resistance vessels, hypertension, and death of transgenic mice, whereas wild-type siblings developed hypertension but did not show increased mortality. L-NAME-treated polyglobulic mice revealed acute left ventricular dilatation and vascular engorgement associated with pulmonary congestion and hemorrhage. In conclusion, we here unequivocally demonstrate that endothelial NO maintains normotension, prevents cardiovascular dysfunction, and critically determines survival in vivo under conditions of increased hematocrit. PMID- 11027360 TI - The osmolyte xylitol reduces the salt concentration of airway surface liquid and may enhance bacterial killing. AB - The thin layer of airway surface liquid (ASL) contains antimicrobial substances that kill the small numbers of bacteria that are constantly being deposited in the lungs. An increase in ASL salt concentration inhibits the activity of airway antimicrobial factors and may partially explain the pathogenesis of cystic fibrosis (CF). We tested the hypothesis that an osmolyte with a low transepithelial permeability may lower the ASL salt concentration, thereby enhancing innate immunity. We found that the five-carbon sugar xylitol has a low transepithelial permeability, is poorly metabolized by several bacteria, and can lower the ASL salt concentration in both CF and non-CF airway epithelia in vitro. Furthermore, in a double-blind, randomized, crossover study, xylitol sprayed for 4 days into each nostril of normal volunteers significantly decreased the number of nasal coagulase-negative Staphylococcus compared with saline control. Xylitol may be of value in decreasing ASL salt concentration and enhancing the innate antimicrobial defense at the airway surface. PMID- 11027361 TI - RhoB prenylation is driven by the three carboxyl-terminal amino acids of the protein: evidenced in vivo by an anti-farnesyl cysteine antibody. AB - Protein isoprenylation is a lipid posttranslational modification required for the function of many proteins that share a carboxyl-terminal CAAX motif. The X residue determines which isoprenoid will be added to the cysteine. When X is a methionine or serine, the farnesyl-transferase transfers a farnesyl, and when X is a leucine or isoleucine, the geranygeranyl-transferase I, a geranylgeranyl group. But despite its CKVL motif, RhoB was reported to be both geranylgeranylated and farnesylated. Thus, the determinants of RhoB prenylation appear more complex than initially thought. To determine the role of RhoB CAAX motif, we designed RhoB mutants with modified CAAX sequence expressed in baculovirus-infected insect cells. We demonstrated that RhoB was prenylated as a function of the three terminal amino acids, i.e., RhoB bearing the CAIM motif of lamin B or CLLL motif of Rap1A was farnesylated or geranylgeranylated, respectively. Next, we produced a specific polyclonal antibody against farnesyl cysteine methyl ester allowing prenylation analysis avoiding the metabolic labeling restrictions. We confirmed that the unique modification of the RhoB CAAX box was sufficient to direct the RhoB distinct prenylation in mammalian cells and, inversely, that a RhoA-CKVL chimera could be alternatively prenylated. Moreover, the immunoprecipitation of endogenous RhoB from cells with the anti farnesyl cysteine antibody suggested that wild-type RhoB is farnesylated in vivo. Taken together, our results demonstrated that the three last carboxyl amino acids are the main determinants for RhoB prenylation and described an anti-farnesyl cysteine antibody as a useful tool for understanding the cellular control of protein farnesylation. PMID- 11027362 TI - Rice dwarf mutant d1, which is defective in the alpha subunit of the heterotrimeric G protein, affects gibberellin signal transduction. AB - Previously, we reported that the rice dwarf mutant, d1, is defective in the alpha subunit of the heterotrimeric G protein (Galpha). In the present study, gibberellin (GA) signaling in d1 and the role of the Galpha protein in the GA signaling pathway were investigated. Compared with the wild type, GA induction of alpha-amylase activity in aleurone cells of d1 was greatly reduced. Relative to the wild type, the GA(3)-treated aleurone layer of d1 had lower expression of Ramy1A, which encodes alpha-amylase, and OsGAMYB, which encodes a GA-inducible transcriptional factor, and no increase in expression of Ca(2 +)-ATPase. However, in the presence of high GA concentrations, alpha-amylase induction occurred even in d1. The GA sensitivity of second leaf sheath elongation in d1 was similar to that of the wild type in terms of dose responsiveness, but the response of internode elongation to GA was much lower in d1. Furthermore, Os20ox expression was up-regulated, and the GA content was elevated in the stunted internodes of d1. All these results suggest that d1 affects a part of the GA-signaling pathway, namely the induction of alpha-amylase in the aleurone layer and internode elongation. In addition, a double mutant between d1 and another GA-signaling mutant, slr, revealed that SLR is epistatic to the D1, supporting that the Galpha protein is involved in GA signaling. However, the data also provide evidence for the presence of an alternative GA-signaling pathway that does not involve the Galpha protein. It is proposed that GA signaling via the Galpha protein may be more sensitive than that of the alternative pathway, as indicated by the low GA responsiveness of this Galpha-independent pathway. PMID- 11027363 TI - Coordinated plant defense responses in Arabidopsis revealed by microarray analysis. AB - Disease resistance is associated with a plant defense response that involves an integrated set of signal transduction pathways. Changes in the expression patterns of 2,375 selected genes were examined simultaneously by cDNA microarray analysis in Arabidopsis thaliana after inoculation with an incompatible fungal pathogen Alternaria brassicicola or treatment with the defense-related signaling molecules salicylic acid (SA), methyl jasmonate (MJ), or ethylene. Substantial changes (up- and down-regulation) in the steady-state abundance of 705 mRNAs were observed in response to one or more of the treatments, including known and putative defense-related genes and 106 genes with no previously described function or homology. In leaf tissue inoculated with A. brassicicola, the abundance of 168 mRNAs was increased more than 2.5-fold, whereas that of 39 mRNAs was reduced. Similarly, the abundance of 192, 221, and 55 mRNAs was highly (>2.5 fold) increased after treatment with SA, MJ, and ethylene, respectively. Data analysis revealed a surprising level of coordinated defense responses, including 169 mRNAs regulated by multiple treatments/defense pathways. The largest number of genes coinduced (one of four induced genes) and corepressed was found after treatments with SA and MJ. In addition, 50% of the genes induced by ethylene treatment were also induced by MJ treatment. These results indicated the existence of a substantial network of regulatory interactions and coordination occurring during plant defense among the different defense signaling pathways, notably between the salicylate and jasmonate pathways that were previously thought to act in an antagonistic fashion. PMID- 11027364 TI - Genetics and visual attention: selective deficits in healthy adult carriers of the epsilon 4 allele of the apolipoprotein E gene. AB - The epsilon4 allele of the apolipoprotein E (APOE) gene is associated with altered brain physiology in healthy adults before old age, but concomitant deficits in cognition on standardized tests of cognitive function have not been consistently demonstrated. We hypothesized that sensitive and specific assessment of basic attentional functions that underlie complex cognition would reveal evidence of impairment in otherwise asymptomatic individuals. We found that as early as middle age, nondemented carriers of the varepsilon4 allele of the APOE gene showed deficits when visual attention was spatially directed by cues in tasks of visual discrimination and visual search, in comparison to those without the epsilon4 allele (epsilon2 and epsilon3 carriers). Two component attentional operations were selectively affected: (i) shifting spatial attention following invalid location cues, and (ii) adjusting the spatial scale of attention during visual search. These changes occurred only in the presence of the epsilon4 allele and without decline in other aspects of attention (vigilance), memory, or general cognition. The results show that specific components of visual attention are affected by APOE genotype and that the course of cognitive aging is subject to selective alteration by a genetic trait. PMID- 11027365 TI - Amblyopes see true alignment where normal observers see illusory tilt. AB - Amblyopia ("lazy eye") is an impairment in visual acuity resulting from abnormal neural development in the visual cortex. We tested the responses of ten amblyopic and six normal observers to illusions of perceived orientation in textures of Gabor patches: the "Fraser illusion," the "phase illusion," and a "tilted chain" illusion. The illusory tilt of the stimulus rows was matched by actual tilt in the opposite direction by using the method of constant stimuli. Amblyopes showed a significant increase in the Fraser illusion, a decrease in the phase illusion, and a reversal of the tilted chain illusion. Amblyopic performance could be simulated in normal observers by reducing the length of the rows. These results can be modeled by a theory which places the neural abnormality in amblyopia at the level of second stage grouping processes. Additionally, the illusions might be useful in the early diagnosis of amblyopia without the need for prior refractive correction. PMID- 11027371 TI - Have a manuscript to publish? why not send it here? PMID- 11027372 TI - Changing prevalence of human immunodeficiency virus-associated oral lesions. PMID- 11027373 TI - Sublingual gland is origin of cervical extravasation mucocele. PMID- 11027374 TI - In reply PMID- 11027375 TI - A review of perioperative corticosteroid use in dentoalveolar surgery. AB - OBJECTIVES: Dental surgeons are often advised to use corticosteroids during and after third molar removal and other dentoalveolar surgery to reduce postsurgical edema, but recommendations for use are rarely accompanied by definitive guidance regarding the type of steroid, dosage, or duration of administration. Many regimens in use appear to be based on anecdotal information from articles in the 1960s and 1970s and might be subtherapeutic. Few regimens have been updated with data from more recent studies, and well-designed comparison studies are lacking. STUDY DESIGN: In this article, the literature from the past 30 years is reviewed, meaningful findings are highlighted, and available data are used as a basis for formulating interim clinical recommendations for corticosteroid use pending the emergence of more evidence-based data. A meta-analysis of data was not performed. RESULTS: Recent data suggest that perioperative corticosteroid regimens should be administered in higher doses and for longer durations than recommended in the past and should be started before surgery for optimum benefit. CONCLUSIONS: Based on the literature review, interim recommendations for the use of corticosteroids are proposed, including dosages and regimens that appear rational for oral, intramuscular, or intravenous corticosteroid administration before and after extractions and other dentoalveolar surgery. These largely empiric recommendations might require adjustment when evidence-based data become available in future studies. There is a great need for well-designed clinical research to further evaluate protocols for corticosteroid use. PMID- 11027376 TI - A comparison of the stability of miniplate with bicortical screw fixation after sagittal split setback. AB - OBJECTIVE: The objective of this study was to decide whether use of bicortical screw fixation provides sufficient stability to dispense with intermaxillary fixation. STUDY DESIGN: Thirty consecutive patients who had undergone surgical setback of the mandible by means of bilateral sagittal split ramus osteotomies were studied. Group 1 (15 patients) had miniplate fixation with intermaxillary fixation for 6 weeks, and group 2 (15 patients) had bicortical screw fixation and immediate postoperative function. The 2 groups were evaluated radiographically for postsurgical changes of pogonion in the early (6 weeks) phase. RESULTS: The results showed that there were no significant differences between the 2 groups. Overall, there was good stability in both groups. CONCLUSION: The use of bicortical screw fixation after sagittal split setback of the mandible provides sufficient stability to dispense with intermaxillary fixation. PMID- 11027377 TI - Healing process after alveolar ridge distraction in sheep. AB - OBJECTIVE: Mandibular augmentation by distraction of the alveolar ridge has been in use for several years. Since 1996, a distraction device that remains in the alveolar ridge after distraction has been used by the Department of Oral and Maxillofacial Surgery of Graz University. The distraction device is transformed into a dental implant after the end of the distraction process and can later be used for prosthetic purposes. We aimed to show the application of the device in this animal experiment and to follow the osseous healing process. STUDY DESIGN: Two distraction implants were inserted into the mandibles of 8 sheep. Distraction of 0.5 mm per day was carried out for 8 days. Two sheep were killed 1, 2, 3, and 6 months after distraction, and the dissected mandibles were examined clinically, radiologically, and histologically. RESULTS: After the first month of distraction, only slight radiopacity of the distraction gap was found. This increased steadily up to the third month after distraction. The boundary between the distraction fragments and the gap disappeared gradually. Six months after distraction, a homogeneous fine-meshed spongiosa structure was found in the area of distraction. Histologic examination showed desmoid ossification in the distraction gap and a continued increase in osteoid. After 6 months, mature bone was found. Only in the center was the rebuilding process not complete. Six months after distraction, osseointegration of the implants was shown in the region of the screw thread and distraction cylinder. CONCLUSIONS: The healing process corresponded to that found in long bones, but showed only desmal ossification. Osseous integration of the distraction implants was found 6 months after distraction, although the implants were stable 3 months after distraction. PMID- 11027378 TI - Oral health management implications in patients with tuberous sclerosis. AB - We report 6 cases of patients with tuberous sclerosis and concomitant dental pathosis. The multiple manifestations in tuberous sclerosis determine its impact on dental therapy. A lack of awareness of this condition may compound the possible side effects of dental treatment. Possible preventive measures by dentists are highlighted in this presentation. Medical evaluation and the control of risk factors in relation to general anesthesia and sedation are key considerations for the management of patients with tuberous sclerosis. PMID- 11027379 TI - Unusual gingival presentation of post-transplantation lymphoproliferative disorder: a case report and review of the literature. AB - Post-transplantation lymphoproliferative disorder is a well-documented complication of solid organ or bone marrow transplantation. Histologically, it is characterized by an abnormal proliferation of lymphocytes, which can range from benign B-cell hyperplasia to malignant lymphoma. Non-Hodgkin's lymphoma (NHL) is associated with several risk factors, such as congenital or acquired immunodeficiency states, autoimmune disorders, and infectious agents (eg, Epstein Barr virus). Primary sites of presentation in the head and neck are Waldeyer's ring, paranasal sinuses, salivary glands, the oral cavity, and the larynx. Clinical appearance of gingival NHL varies but is usually found to be an asymptomatic gingival enlargement or mass resembling a pyogenic granuloma. We present a patient with a gingival ulceration that was subsequently diagnosed as Epstein-Barr virus malignant lymphoma resulting from the immunosuppression needed to prevent graft-versus-host disease after bone marrow transplantation. PMID- 11027380 TI - Temporomandibular joint pain: relationship to internal derangement type, osteoarthrosis, and synovial fluid mediator level of tumor necrosis factor-alpha. AB - OBJECTIVES: The purpose of this study was to investigate whether patients with temporomandibular joint (TMJ)-related pain classified as capsulitis/synovitis may be linked to magnetic resonance imaging (MRI) findings of internal derangement, osteoarthrosis, or the synovial fluid aspirate findings of tumor necrosis factor alpha (TNF-alpha) level. STUDY DESIGN: The study comprised 23 patients with temporomandibular disorders (TMD), who had nonchronic pain (pain onset < or =6 months) and a unilateral TMJ-related diagnosis of capsulitis/synovitis. Bilateral sagittal and coronal magnetic resonance images were obtained to establish the presence or absence of internal derangement, osteoarthrosis, or both. TMJ synovial fluid aspirates were obtained from the pain and contralateral nonpain sides to determine the TNF-alpha level. RESULTS: Comparison of the TMJ side related data showed a significant relationship between the clinical TMD diagnosis of capsulitis/synovitis and the MRI diagnoses of TMJ internal derangement (P =.002) and of TMJ internal derangement type (P =.04). The mean TNF-alpha level in synovial fluid aspirates from TMJs assigned a clinical TMD diagnosis of capsulitis/synovitis was significantly higher than in those obtained from contralateral nonpain sides (P =.001). There was no correlation between the clinical diagnosis of capsulitis/synovitis and the MRI diagnosis of TMJ osteoarthrosis (P =.13) or between the MRI diagnosis of TMJ osteoarthrosis and that of TMJ internal derangement (P =.70) or TMJ internal derangement type (P =.33). CONCLUSIONS: The results suggest that the TMJ pain condition of capsulitis/synovitis is related to TMJ-side specific MRI diagnoses of internal derangement and internal derangement type, and synovial fluid aspirate findings of TNF-alpha level. The data confirm the concept of elevated mediator level as a diagnostic approach for patients presenting with TMJ-related pain. MRI and synovial fluid aspirates may be used as diagnostic methods for evaluating TMJ related pain conditions. PMID- 11027381 TI - Primary non-Hodgkin's lymphoma of the mandible treated with radiotherapy, chemotherapy, and autologous peripheral blood stem cell transplantation. AB - Extranodal presentation in non-Hodgkin's lymphoma (NHL) is uncommon, and the mandible is very rarely involved. Primary NHL of the mandible, for the most part, has intermediate or high malignancy and has a much greater incidence of local recurrence compared with other sites of involvement. A 48-year-old Japanese man with NHL of the mandible received radiotherapy, followed by high-dose chemotherapy supported with peripheral blood stem cell transplantation (PBSCT). High-dose cyclophosphamide, Adriamycin, and vincristine were used for pretransplant conditioning. He achieved complete remission and has survived in continuous complete remission for more than 72 months to date. Marrow-ablative chemotherapy facilitated by PBSCT is thought to be useful as part of the primary therapy for patients with NHL who have poorer prognoses. PMID- 11027382 TI - Painful trigeminal neuropathy caused by severe mandibular resorption and nerve compression in a patient with systemic sclerosis: case report and literature review. AB - Systemic sclerosis is a multi-system disorder characterized by abundant fibrosis of the skin, blood vessels, and visceral organs. Although resorption of the mandible has been commonly observed and reported, we found no report of resorption leading to a painful neuropathy of the inferior alveolar nerve. We report a case of a patient with systemic sclerosis, severe resorption of the angles, and inferior alveolar border of the mandible, resulting in a compression neuropathy of the inferior alveolar nerve. Diagnostic tests, medical treatment, and surgical treatment are discussed, and the relevant literature is reviewed. PMID- 11027383 TI - Epithelial-myoepithelial carcinoma arising in pleomorphic adenoma of the palate. AB - A case of epithelial-myoepithelial carcinoma (EMC) in pleomorphic adenoma (PA) occurring in the palate of a 72-year-old woman is reported. The tumor was composed of 2 different components, PA and EMC, accounting for approximately 40% and 60% of the whole tumor, respectively. The EMC showed multiple tubular or solid nests, which were separated by a basement membrane and consisted of variable proportions of 2 cell types, cuboidal epithelial cells positive for cytokeratin and clear myoepithelial cells positive for glial fibrillary acid protein, whereas the myoepithelial nests of PA intermingled with hyaline and myxoid stroma. The malignancy was demonstrated by convincing evidence of invasion into the submucosa, although the EMC component was mostly surrounded by the PA components. An increased immunoreactivity of proliferating cell nuclear antigen in the EMC area in comparison to the PA area also suggested EMC arising in a PA. PMID- 11027384 TI - Oral bowenoid lesions: differential diagnosis and pathogenetic insights. AB - OBJECTIVE: To determine if oral lesions exhibiting bowenoid features reflect the diverse microscopic appearance and biologic behaviour of Bowen's disease and bowenoid papulosis of the skin and genitalia. STUDY DESIGN: Seven cases of oral bowenoid lesions (6 with follow-up data) were assessed for differences in histologic features, human papillomavirus (HPV) viral status, and selected immunohistochemically detectable cell cycling proteins (p53, WAF-1, Cyclin D1, Bcl-2) and were correlated with available follow-up data. RESULTS: Two histologic subsets were identified. One, which was believed to correspond to Bowen's disease, exhibited large numbers of transepithelial apoptotic bodies, dyskeratotic cells and mitoses (bowenoid elements), poor differentiation of background epithelial cells, and consistent HPV-16/18 positivity. The other, believed to correspond to bowenoid papulosis, exhibited few bowenoid elements, good background differentiation, and inconsistent HPV-16/18 positivity. One of the aggressive cases exhibited repeated recurrences despite apparent total clinical excision, whereas none of the other group recurred. CONCLUSION: Although a small number of cases are in this study, results suggest that oral bowenoid lesions may exhibit histopathologic and behavioral variations ranging from oral Bowen's disease to oral bowenoid papulosis. Studies on more cases are needed to confirm this initial impression. PMID- 11027385 TI - Large granular lymphocyte leukemia and its association with oral neutropenic ulcerations: a case report. AB - Large granular lymphocyte leukemia is a rare chronic indolent disorder commonly associated with severe neutropenia. The pathogenesis of the neutropenia is unclear. A case is presented of a 74-year-old man who had recurrent oral ulcerations for over a year before a diagnosis was made. These recurrent oral ulcers cleared with treatment and have not returned. The differential diagnosis of persistent oral ulcerations includes trauma; viral, fungal, and bacterial infections; systemic disease; or various malignant conditions. The oral ulcers in this man were likely infectious in nature and related to the severe chronic neutropenia. This case serves to illustrate the potentially complex nature of oral ulcers. PMID- 11027386 TI - Mucocele of the anterior lingual salivary glands (glands of Blandin and Nuhn): report of 5 cases. AB - The anterior lingual salivary glands (glands of Blandin and Nuhn) are mixed mucous and serous glands that are embedded within the musculature of the anterior tongue ventrum. Five cases of mucocele of the glands of Blandin and Nuhn are presented. These mucoceles on the anterior tongue ventrum were exophytic and resembled pyogenic granulomata, polyps, or squamous papillomata. In 2 cases, the onset of the mucocele was associated with trauma to the anterior tongue. All cases were mucus extravasation phenomena. A history of trauma and recovery of mucus with fine needle aspiration are helpful in the clinical diagnosis of mucocele of the glands of Blandin and Nuhn, as are the following characteristics of the mucocele: rapid onset, increase and reduction in size, bluish color, and fluid-filled consistency. During surgery, the glands that are deep in the tongue musculature are commonly left behind, resulting in persistence of the lesion. Careful clinical evaluation of these lesions and preoperative awareness of the surgical anatomy of the glands of Blandin and Nuhn may minimize the need for repeated surgical procedures. PMID- 11027387 TI - Segmental odontomaxillary dysplasia: a case report and review of the literature. AB - Segmental odontomaxillary dysplasia (SOD) is a rare, unilateral developmental disorder of the maxilla involving abnormal growth and maturation of the bone, lack of one or both premolars, altered primary molar structure, delayed tooth eruption, and fibrous hyperplasia of the gingiva. In this, the twenty-third reported case of SOD, the literature is reviewed, and the clinical, radiographic, and histopathologic data are described. Computed tomographic scans of this case showed that the involved segment of the maxilla extends mesiodistally from the permanent cuspid to the mesial portion of the first permanent molar, largely limited to the area of the missing premolars. However, the affected bone extends superiorly in the lateral wall of the maxilla to the zygoma and base of the orbit. This article is intended to serve as baseline data for a future article, describing the natural history and possible treatment of SOD, which remain undocumented. PMID- 11027388 TI - A comparison of stainless steel and nickel-titanium H-type instruments of identical design: torsional and bending tests. AB - OBJECTIVE: For a true comparison of clinically relevant qualities of instruments manufactured from different alloys, the instrument must be identical in design. The purpose of this study was to perform mechanical tests on stainless steel and nickel-titanium instruments of identical design. STUDY DESIGN: Identical experimental endodontic files of H-type with double helix were fabricated from stainless steel and nickel-titanium alloys. Torsional and bending tests were carried out according to American National Standards Institute/American Dental Association specification No. 58 and International Standards Organization No. 3630/1. Changes in instruments during the testing were also analyzed with scanning electron microscope technique. RESULTS: The results indicated that the torsional moment of the stainless steel files was significantly higher than for the nickel-titanium files (P <.001), whereas the mean angular deflection for the nickel-titanium files was significantly higher. The mean bending moment for the nickel-titanium files was significantly lower (P <.001) than the mean value for the stainless steel files. The scanning electron microscope demonstrated that the fracture surfaces of the stainless steel files were brittle, whereas the nickel titanium files had a ductile fracture. CONCLUSIONS: When the design of endodontic instruments of different alloys is identical, the nickel-titanium files are more flexible in bending. However, nickel-titanium files require less force to deform to fracture. PMID- 11027389 TI - Effect of trephination on postoperative pain and swelling in symptomatic necrotic teeth. AB - OBJECTIVE: The purpose of this prospective, randomized, blinded study was to determine the effect of trephination on postoperative pain and swelling in symptomatic necrotic teeth. STUDY DESIGN: Fifty emergency patients participated, and each had a clinical diagnosis of a symptomatic necrotic tooth with associated periapical radiolucency. After endodontic treatment, patients randomly received either a trephination or mock trephination procedure. The trephination procedure used an intraosseous perforator to provide an initial opening in the cortical bone that was enlarged with files (No. 25 through No. 70) and an endodontic spoon. After surgery, each patient received ibuprofen; acetaminophen with codeine (30 mg); and a 7-day diary to record pain, percussion pain, swelling, and number and type of pain medication taken. RESULTS: The majority of patients with symptomatic necrotic teeth had significant postoperative pain and required analgesics to manage this pain. The use of a trephination procedure with an intraosseous perforator, files, and a spoon excavator did not significantly reduce pain, percussion pain, swelling, or the number of analgesic medications taken in symptomatic necrotic teeth with periapical radiolucencies (P >.05). CONCLUSION: We cannot recommend the routine use of a trephination procedure, as used in this study, for relief of pain in symptomatic necrotic teeth with radiolucencies. PMID- 11027390 TI - Tumor necrosis factor-alpha and transforming growth factor-beta1 in chronic periapical lesions. AB - OBJECTIVE: Proinflammatory cytokines are involved in the pathogenesis of periapical lesions. The purpose of this study was to evaluate the presence of the cytokines tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor beta(1) (TGF-beta(1)) in periapical pathosis and to determine their relationship to the size of the lesions. STUDY DESIGN: One tooth from each of 25 patients was root-end resected, and the periapical lesion was collected. The amounts of TNF alpha and TGF-beta(1) were assessed by enzyme-linked immunosorbent assay. RESULTS: TGF-beta(1) was detected in 21 of 25 lesions. In samples with scar tissue, no TGF-beta(1) activity was detected. A statistically significant correlation was found between TGF-beta(1) per milligram of tissue and the diameter of the lesions. TNF-alpha was detected in only 2 samples. CONCLUSIONS: TGF-beta(1) was present in periapical granulomas and cysts but not in lesions with scar tissue. The correlation between the amount of TGF-beta(1) per milligram of tissue and the size of the lesion was significant. PMID- 11027391 TI - Case-control study on self-reported osteoporotic fractures and mandibular cortical bone. AB - OBJECTIVES: The purpose of this case-control study was to determine whether the radiographic appearance of the mandibular cortical bone in patients who were elderly and noninstitutionalized was related to a self-reported history of osteoporotic fractures. STUDY DESIGN: Patients who had a billing statement at the School of Dentistry dated between 1993 and 1996, who were older than 60, and who had a panoramic radiograph were invited to be interviewed regarding fracture history (circumstances and year of fracture) and risk factors for osteoporosis. Cases (n = 93) were individuals reporting osteoporotic fractures (fractures occurring after minor impact). Controls (n = 394) were individuals reporting traumatic fractures (n = 105) or no fractures (n = 289). Blinded to case-control status, we evaluated the mandibular cortex on a panoramic radiograph and classified them as normal (even and sharp endosteal margin), moderately eroded (evidence of lacunar resorption or endosteal cortical residues), or severely eroded (unequivocal porosity). In addition, cortical thickness was measured below the mental foramen. RESULTS: After adjustment for potentially confounding factors, the odds ratio for an osteoporotic fracture associated with moderately eroded and severely eroded mandibular cortices was 2.0 (95% CI, 1.2 to 3.3) and 8.0 (95% CI, 2.0 to 28.9), respectively. After adjusting for all potentially confounding factors, we found that the cortex was 0.54 mm (or 12%) thinner in subjects with an osteoporotic fracture compared with controls (95% CI, 0.25 to 0.84 mm). CONCLUSIONS: Subjects with a self-reported history of osteoporotic fractures tend to have increased resorption and thinning of the mandibular lower cortex. PMID- 11027392 TI - Lymphoscintigraphy for interpretation of changes of cervical lymph node function in patients with oral malignant tumors: comparison of Tc-99m-Re and Tc-99m-HSA-D. AB - OBJECTIVE: The purpose of this study is to compare the usefulness of technetium 99m-rhenium colloid (Tc-99m-Re) and technetium-99m-human serum albumin diethylene triamine-pentaacetic acid (Tc-99m-HSA-D) as lymphoscintigraphic agents and to discuss the significance of lymphoscintigraphy in comparison with computed tomography (CT), magnetic resonance imaging (MRI), and ultrasonography (US). STUDY DESIGN: Dynamic and static lymphoscintigraphies were performed with Tc-99m Re and Tc-99m-HSA-D. The usefulness of the 2 agents was evaluated statistically in comparison with pathologic findings and palpation. The significance of lymphoscintigraphy is discussed in comparison with CT, MRI, and US (by the literature of CT, MRI, and US). RESULTS: Lymphoscintigraphy was superior to palpation, and Tc-99m-Re was superior to Tc-99m-HSA-D in accuracy, specificity, and the incidence of true-positive and false-positive. Statistical significance was shown between the static lymphoscintigraphy with Tc-99m-Re and palpation. The reliability of lymphoscintigraphy seemed to be slightly inferior to CT and MRI in specificity and accuracy. However, lymphoscintigraphy had some advantages that CT and MRI lacked; for example, it showed very high sensitivity (100%) and no false negative (0%). It also showed changes of lymph node function, showed all levels of neck nodes at one time, and showed a possibility of detecting small lymph node metastases. CONCLUSION: Lymphoscintigraphy was more useful than palpation in detecting lymph node metastases, and Tc-99m-Re was superior to Tc-99m-HSA-D as an agent. Lymphoscintigraphy is significant when it is performed as a preliminary examination before CT or MRI. PMID- 11027393 TI - Effective dose equivalent and effective dose: comparison for common projections in oral and maxillofacial radiology. AB - OBJECTIVES: Effective dose equivalents (H(E)) and effective doses (E) for radiographic projections common in dentistry, calculated from the same organ dose distributions, are presented to determine whether the 2 quantities can be directly compared. STUDY DESIGN: Doses to all organs and tissues in the head, neck, trunk, and proximal extremities were determined for each projection (intraoral full-mouth radiographic survey, panoramic, cephalometric, temporomandibular tomograms, and submentovertex view) by computer simulation with Monte Carlo methods. H(E) and E were calculated from these complete distributions and by methods prescribed by the International Commission on Radiological Protection (ICRP). RESULTS: H(E) and E computed from complete dose distributions were found comparable within a few percentage points. However, those computed by strict application of ICRP methods were not. CONCLUSION: For radiographic projections with highly localized dose distributions, such as those common in dentistry, direct comparison of H(E) and E may not be meaningful, unless both computation algorithms are known. PMID- 11027394 TI - Long-standing intranasal foreign body: an incidental finding on dental radiograph: a case report and literature review. AB - A case of an incidental finding of an intranasal foreign body on a dental panoramic radiograph is reported. The foreign body, a calculus-encrusted plastic pearl, had been asymptomatic despite being present for over 20 years. The diagnosis of a nasal foreign body in a patient is of significance if an oral surgical procedure is being planned with the patient under general anesthesia because the foreign body may be dislodged into the airway during nasal intubation. Similar reports of incidentally discovered intranasal foreign bodies on dental radiographs are reviewed. The epidemiology, diagnosis, management, and complications of nasal foreign bodies are discussed. PMID- 11027395 TI - Spatial sensitivity of macaque inferior temporal neurons. AB - Recent findings in dorsal visual stream areas and computational work raise the question whether neurons at the end station of the ventral visual stream can code for stimulus position. The authors provide the first detailed, quantitative data on the spatial sensitivity of neurons in the anterior part of the inferior temporal cortex (area TE) in awake, fixating monkeys. They observed a large variation in receptive field (RF) size (ranging from 2.8 degrees to 26 degrees ). TE neurons differed in their optimal position, with a bias toward the foveal position. Moreover, the RF profiles of most TE neurons could be fitted well with a two-dimensional Gaussian function. Most neurons had only one region of high sensitivity and showed a smooth decline in sensitivity toward more distal positions. In addition, the authors investigated some of the possible determinants of such spatial sensitivity. First, testing with low-pass filtered versions of the stimuli revealed that the general preference for the foveal position and the size of the RFs was not due simply to TE neurons receiving input with a lower spatial resolution at more eccentric positions. The foveal position was still preferred after intense low-pass filtering. Second, although an increase in stimulus size consistently broadened spatial sensitivity profiles, it did not change the qualitative features of these profiles. Moreover, size selectivity of TE neurons was generally position invariant. Overall, the results suggest that TE neurons can code for the position of stimuli in the central region of the visual field. PMID- 11027396 TI - Differential effect of thyroid hormone deficiency on the growth of calretinin expressing neurons in rat spinal cord and dorsal root ganglia. AB - The development of spinal cord or dorsal root ganglia neurons expressing calretinin (CR) was studied in thyroid hormone-deficient rats. Immunocytochemical and morphometric analyses showed that the hypothyroidism induced a significant decrease in the number and size of immunoreactive neurons in the spinal cord, as well as stunted growth and arborization of the axons and dendrites. These alterations were observed at different embryonic ages and persisted during the whole postnatal life. In adult hypothyroid rats, the mean number of CR-positive neurons per spinal cord section (31.2 +/- 2.3 in laminae I and II and 30.5 +/- 5.5 in laminae III-X) was significantly decreased (P < 0.001 and P = 0.024, respectively) compared with adult normal rats (68.7 +/- 8.9 and 50.0 +/- 11.0, respectively). In the peripheral nervous system, hypothyroidism altered the growth of sensory neurons expressing CR protein mainly during embryonic life. In comparison with normal rats, hypothyroid embryonic animals showed not only reduced cell size but also a significantly decreased percentage of CR-positive neurons (6.6 +/- 0. 9% in normal, 2.1 +/- 0.3% in hypothyroid rats, P < 0.001). In contrast, although the size of neurons was reduced in hypothyroid young and adult rats, there was no reduction in the percentage of CR-positive neurons. These results showed that thyroid hormone deficiency altered differentially the development of neurons expressing CR protein in the central and peripheral nervous systems. This suggests that central and peripheral neurons are heterogeneous in their sensitivity to thyroid hormone. PMID- 11027397 TI - Age-related decreases in GTP-cyclohydrolase-I immunoreactive neurons in the monkey and human substantia nigra. AB - Guanosine triphosphate cyclohydrolase I (GTPCHI) is a critical enzyme in catecholamine function and is rate limiting for the synthesis of the catecholamine co-factor tetrahydrobiopterin. The present study assessed the distribution of GTPCHI immunoreactivity (-ir) within the monkey and human ventral midbrain and determined whether its expression is altered as a function of age. Light and confocal microscopic analyses revealed that young monkeys and humans displayed GTPCHI-ir within melanin-containing and tyrosine-hydroxylase-ir neurons in primate substantia nigra. Stereological counts revealed that there was a 67.4% reduction in GTPCHI-ir neuronal number, a 63.5% reduction in GTPCHI-ir neuronal density, and a 37.6% reduction in neuronal volume in aged monkeys relative to young cohorts. Similar age-related changes were seen in humans, in whom there were significant reductions in the number of GTPCHI-ir nigral neurons in middle age (58.4%) and aged (81.5%) cases relative to young cohorts. The density of GTPCHI-ir neurons within the nigra was similarly reduced in middle-aged (63.0%) and aged (81.8%) cases. In contrast to monkeys, aged humans did not display shrinkage in the volume of GTPCHI-ir nigral neurons. The presence of numerous melanin-positive, but GTPCHI-ir immunonegative, neurons in the aged monkey and human nigra indicates that these decreases represent an age-related phenotypic downregulation of this enzyme and not a loss of neurons per se. These data indicate that there is a dramatic decrease in GTPCHI-ir in nonhuman primates and humans as a function of age and that loss of this enzyme may be partly responsible for the age-related decrease in dopaminergic tone within nigrostriatal systems. PMID- 11027398 TI - Immunohistochemical studies of the localization of neurons containing the enzyme that synthesizes dopamine, GABA, or gamma-hydroxybutyrate in the rat substantia nigra and striatum. AB - gamma-Hydroxybutyrate (GHB) is an endogenous metabolite of gamma-aminobutyric acid (GABA), which is synthesized in the neuronal compartment of the central nervous system. This substance possesses several properties that support its role as a neurotransmitter/neuromodulator in brain. In particular, it is synthesized by a specific pathway that transforms GABA into succinic semialdehyde via GABA-T activity; then succinic semialdehyde is converted into GHB by a specific succinic semialdehyde reductase (SSR). The last enzyme is considered as a marker for neurons that synthesize GHB. This compound binds in brain to receptors whose distribution, ontogenesis, kinetics, and pharmacology are specific. Endogenous GHB, but also GHB exogenously administered to rats, participate in the regulation of dopaminergic activity of the nigrostriatal pathway. To investigate the distribution of GHB neurons in this pathway and the anatomic relationships between dopaminergic and GHB neurons, immunocytochemical identification of dopamine, GABA, and GHB neurons was carried out in the substantia nigra and striatum of the rat. The following markers for these neurons were used: anti tyrosine hydroxylase (TH) antibodies for dopamine neurons, anti-glutamate decarboxylase (GAD) antibodies for GABA neurons, and anti-succinic semialdehyde reductase (SSR) antibodies for GHB neurons. GABA neurons were studied because GAD and SSR co-exist frequently in the same neuron, and GABA alone also exerts its own regulatory effects on dopaminergic neurons. This study reveals the co existence of GAD/SSR and GAD/SSR/TH in numerous neurons of the substantia nigra. However, some neurons appear to be only GAD or SSR positive. In the striatum, TH positive terminals surround many GHB neurons. GAD innervation is abundant in close contact with unlabeled neurons in the caudate-putamen, whereas distinct SSR positive punctuates are also present. The existence of SSR-reactive synapses and neurons was confirmed in the striatum at the electron microscopic level. On the basis of these results, a clear anatomo-functional relationship between GHB and dopamine networks cannot be defined; however, we propose the modulation by GHB of striatal intrinsic neurons that could then interfere with the presynaptic control of dopaminergic activity. PMID- 11027399 TI - Patterns of cell death in mouse anteroventral cochlear nucleus neurons after unilateral cochlea removal. AB - Developmental changes that influence the results of removal of afferent input on the survival of neurons of the anteroventral cochlear nucleus (AVCN) of mice were examined with the hope of providing a suitable model for understanding the cellular and molecular basis for these developmental changes in susceptibility. We performed unilateral cochlear ablation on wild-type mice at a variety of ages around the time of hearing onset to determine developmental changes in the sensitivity of AVCN neurons to afferent deprivation. In postnatal day 5 (P5) mice, cochlea removal resulted in 61% neuronal loss in the AVCN. By age P14, fewer than 1% of AVCN neurons were lost after this manipulation. This reveals a rather abrupt change in the sensitivity to disruption of afferent input, a critical period. We next investigated the temporal events associated with neuron loss after cochlea removal in susceptible animals. We demonstrate that significant cell loss occurs within 48 hours of cochlea removal in P7 animals. Furthermore, evidence of apoptosis was observed within 12 hours of cochlea removal, suggesting that the molecular events leading to cell loss after afferent deprivation begin to occur within hours of cochlea removal. Finally, we began to examine the role of the bcl-2 gene family in regulating afferent deprivation induced cell death in the mouse AVCN. AVCN neurons in mature bcl-2 knockout mice demonstrate susceptibility to removal of afferent input comparable to neonatal sensitivity of wild-type controls. These data suggest that bcl-2 is one effector of cell survival as these cells switch from afferent-dependent to -independent survival mechanisms. PMID- 11027400 TI - Improved immunohistochemical detection of postsynaptically located PSD-95/SAP90 protein family by protease section pretreatment: a study in the adult mouse brain. AB - Postsynaptic density (PSD)-95, SAP102, and Chapsyn-110 are members of the PSD 95/SAP90 protein family, which interact with the C-terminus of N-methyl-D aspartate (NMDA) receptor and shaker-type potassium channel subunits. Here we report that appropriate section pretreatment with pepsin has led to qualitative and quantitative changes in light microscopic immunohistochemical detection of the protein family. First, pepsin pretreatment lowered the concentration of affinity-purified primary antibodies, while it greatly increased the intensity of immunoreactions. Second, the resulting overall distributions of PSD-95, SAP102, and Chapsyn-110 in the adult mouse brain were consistent with their mRNA distributions. Third, instead of the reported patterns of somatodendritic labeling, tiny punctate staining in the neuropil became overwhelming. Fourth, many PSD-95-immunopositive puncta were apposed closely to synaptophysin-positive nerve terminals and overlapped with NMDA receptor subunits. By postembedding immunogold, the PSD-95 antibody was shown to label exclusively the postsynaptic density at asymmetrical synapses. Based on these results, we conclude that antibody access and binding to the postsynaptically located PSD-95/SAP90 protein family are hindered when conventional immunohistochemistry is adopted, and that pepsin pretreatment effectively unmasks the postsynaptic epitopes. On the other hand, PSD-95 in axon terminals of cerebellar basket cells, where high levels of potassium channels are present, was detectable irrespective of pepsin pretreatment, suggesting that PSD-95 antibody is readily accessible to the presynaptic epitopes. Consequently, the present immunohistochemical results have provided light microscopic evidence supporting the prevailing notion that the PSD 95/SAP90 protein family interacts with NMDA receptor subunits and potassium channel subunits. PMID- 11027401 TI - Differential regulation of trophic factor receptor mRNAs in spinal motoneurons after sciatic nerve transection and ventral root avulsion in the rat. AB - After sciatic nerve lesion in the adult rat, motoneurons survive and regenerate, whereas the same lesion in the neonatal animal or an avulsion of ventral roots from the spinal cord in adults induces extensive cell death among lesioned motoneurons with limited or no axon regeneration. A number of substances with neurotrophic effects have been shown to increase survival of motoneurons in vivo and in vitro. Here we have used semiquantitative in situ hybridization histochemistry to detect the regulation in motoneurons of mRNAs for receptors to ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) 1-42 days after the described three types of axon injury. After all types of injury, the mRNAs for GDNF receptors (GFRalpha-1 and c-RET) and the LIF receptor LIFR were distinctly (up to 300%) up-regulated in motoneurons. The CNTF receptor CNTFRalpha mRNA displayed only small changes, whereas the mRNA for membrane glycoprotein 130 (gp130), which is a critical receptor component for LIF and CNTF transduction, was profoundly down-regulated in motoneurons after ventral root avulsion. The BDNF full-length receptor trkB mRNA was up-regulated acutely after adult sciatic nerve lesion, whereas after ventral root avulsion trkB was down-regulated. The NT-3 receptor trkC mRNA was strongly down-regulated after ventral root avulsion. The results demonstrate that removal of peripheral nerve tissue from proximally lesioned motor axons induces profound down-regulations of mRNAs for critical components of receptors for CNTF, LIF, and NT-3 in affected motoneurons, but GDNF receptor mRNAs are up-regulated in the same situation. These results should be considered in relation to the extensive cell death among motoneurons after ventral root avulsion and should also be important for the design of therapeutical approaches in cases of motoneuron death. PMID- 11027402 TI - Distribution of choline acetyltransferase (ChAT) immunoreactivity in the central nervous system of a chondrostean, the siberian sturgeon (Acipenser baeri). AB - All studies to date of cholinergic systems of bony fishes have been done in teleosts. To gain further insight into the evolution of the cholinergic systems of bony fishes, we have studied the brain of a chondrostean fish, the Siberian sturgeon (Acipenser baeri, Brandt), by using an antibody against choline acetyltransferase (ChAT). This study showed the presence of ChAT-immunoreactive (ChAT-ir) neurons in the preoptic region (parvocellular and magnocellular preoptic nuclei and suprachiasmatic nucleus), the periventricular and tuberal hypothalamus, the saccus vasculosus, the dorsal thalamus, and the habenula. The mesencephalic tegmentum contained ChAT-ir cells in the torus semicircularis and torus lateralis. The isthmus contained several cholinergic populations: the nucleus isthmi, the lateral nucleus of the valvula, the secondary visceral nucleus, and the dorsal tegmental nucleus. The motor neurons of the cranial nerves and the spinal motor column were strongly immunoreactive. The medial (sensory) trigeminal nucleus also contained a ChAT-ir neuronal population. The distribution of ChAT-ir neurons in the sturgeon brain showed some notable differences with that observed in teleosts, such as the absence of cholinergic cells in the telencephalon and the optic tectum. Several brain regions were richly innervated by ChAT-ir fibers, particularly the telencephalon, optic tectum, thalamus, posterior tubercle, and interpeduncular nucleus. The hypothalamo-hypophyseal tract, the tract of the saccus vasculosus, the fasciculus retroflexus, and an isthmo-mesencephalo-thalamic tract were the most conspicuous cholinergic bundles. Comparative analysis of these results suggests that teleosts have conserved most traits of the cholinergic system of the sturgeon, having acquired new cholinergic populations during evolution. PMID- 11027403 TI - Developmental changes in the localization of activated C-JUN N-terminal kinase (JNK/SAPK) in the chick spinal cord. AB - To examine the role of c-Jun N-terminal kinase (JNK/SAPK) in the developing nervous system of vertebrates, the localization of an active form of JNK, phosphorylated JNK (p-JNK), was studied in the lumbosacral spinal cord of the chick embryo. We also examined the localization of phosphorylated neurofilaments (NFs, potential targets of p-JNK) and cyclin-dependent kinase 5 (Cdk5), which is known to phosphorylate cytoskeletal proteins, including NFs, and compared their expression with that of p-JNK. Additionally, the localization of phosphorylated forms of c-Jun and ATF-2 was compared with that of p-JNK. On embryonic day 3 (E3), the expression of p-JNK was observed in regions containing early-projecting axons. Axons in these regions also expressed phosphorylated NFs. Subsequently, on E5 and E8, the expression of both p-JNK and phosphorylated NFs increased concomitantly in the axonal tracts in the spinal white matter. Thus, white matter expressed both p-JNK and phosphorylated NFs, whereas there was only weak expression of Cdk5. By E13, the spinal cord expression pattern of p-JNK and phosphorylated NFs had changed compared to earlier ages. Although phosphorylated NFs were still expressed in the white matter, the expression of p-JNK was decreased in axons in the white matter, whereas strong p-JNK expression appeared in cell nuclei in the gray matter. In summary, the present study revealed that the localization of p-JNK in the spinal cord changes dramatically from axons to cell nuclei during development, suggesting multiple roles of p-JNK, depending on the developmental age. PMID- 11027404 TI - Do subplate neurons comprise a transient population of cells in developing neocortex of rats? AB - Studies were undertaken to determine whether neurons of the subplate layer represent a transient or stable population of cells in developing neocortex of rat. The first set of studies sought to determine the fraction of subplate neurons that is lost during early postnatal development. The optical dissector method was used to analyze fluorescently stained material in animals the age of postnatal day 0 (P0) to P40. These results demonstrate a reduction of slightly less than half of the total number of subplate neurons from P0 to P40. Counts of labeled cells in littermates at varied ages after [(3)H]thymidine or BRDU treatment on gestational day 14 (G14 - birthdate of occipital subplate neurons) or G18 (birthdate of layers III-IV neurons) demonstrate loss of approximately 50% of neurons in the subplate layer between P0 and P40, somewhat greater than the loss of neurons from cortical layers III-IV. The second set of studies investigated whether subplate neurons display cellular atrophy during postnatal development. Analysis of subplate neurons injected intracellularly with Lucifer yellow in fixed slice preparations indicates no reduction in soma size, number of dendrites, or extent of dendritic fields of subplate neurons taken from animals age P0 to P60. The third set of studies investigated whether functional markers of subplate neurons are reduced during postnatal development. Analysis of tissue stained histochemically for cytochrome oxidase or acetylcholinesterase, or stained immunocytochemically for GABA, somatostatin, or neuropeptide Y, demonstrate a remarkable loss of expression of staining patterns from late gestational ages to P20. These data demonstrate that, although subplate neurons seem not to be a transient population of cells in the usual sense of being eliminated by cell death or structural atrophy, the loss of histochemical and immunocytochemical markers indicates that they may be a functionally transient population of cells. PMID- 11027406 TI - In memoriam PMID- 11027405 TI - Normal development of the ipsilateral retinocollicular pathway and its disruption in double endothelial and neuronal nitric oxide synthase gene knockout mice. AB - The development of the ipsilateral retinocollicular pathway involves activity dependent refinement in which misdirected axons retract to form a precise retinotopic map in adults. This refinement is altered by disruption of genes for the endothelial and neuronal isoforms of nitric oxide synthase (e,nNOS), but the extent of disruption during early development is not known. Therefore, we studied the refinement of this pathway in normal C57/BL6 and e,nNOS double knockouts from P4 to P21 and in adults. Anterograde tracers were injected into one eye to localize the ipsilateral retinal projection (IRP) within the superior colliculus (SC). At P4, the IRP in normal mice was distributed throughout the dorsoventral extent of the superficial gray layer (SGL) across most of the rostrocaudal axis of SC. Between P4 and P9, the pathway retracted to the rostromedial SC, and retracted further between P15 and P21, such that multiple patches of label were seen only in the rostral 200-300 microm. Refinement also began to occur between P4 and P9 in e,nNOS double knockout mice, but labeling was more extensive in P9, P15, and P21 knockout animals. This delay in refinement was confirmed quantitatively at P15 where differences in the area occupied by the pathway were statistically significant. The refinement process is therefore in progress in both normal and e,nNOS knockout mice before eye opening but is significantly delayed in the double knockouts. The IRP in normal mice is also more exuberant at early ages, and the process of refinement more protracted than has been previously reported, suggesting that there is a prolonged critical period of synaptic plasticity. PMID- 11027407 TI - Estrogens and prostatic disease. International Prostate Health Council Study Group. PMID- 11027408 TI - Prevention of spontaneous prostate-related cancer in Lobund-Wistar rats by a soy protein isolate/isoflavone diet. AB - BACKGROUND: Epidemiological surveys recorded that men in the Orient (Japan and China) consuming diets high in soy food were at low risk of developing clinical prostate cancer, compared to a relatively high risk among men in the West who consumed diets low in soy food. Soybeans contain phytoestrogens (isoflavones) with many recorded anticancer mechanisms. The Lobund-Wistar (L-W) rat is a unique model system: approximately 30% develop metastasizing adenocarcinomas spontaneously in the anterior prostate-seminal vesicle complex (P-SV), from which the tumors expand into the dorsolateral lobes. L-W rats are inherently predisposed, possibly by unusually high levels of circulating testosterone (T), to develop P-SV tumors which are T-dependent in the early stages and T independent in advanced stages of tumorigenesis. METHODS: L-W rats were fed two diets from age 2-24 months: 1) natural ingredient diet L-485 (Harlan TekLad Diets, Madison, WI) containing soy meal, or 2) a modified starch-casein diet in which soy protein isolate/isoflavones (SPII) replaced casein as a source of protein. RESULTS: At age 24 months, 3 of 99 (3%) rats on diet SPII and 30 of 100 (30%) rats on diet L-485 developed spontaneous P-SV cancers. Rats on the SPII diet manifested a significant reduction of circulating T, approaching physiological levels. Failure of the rats on diet L-485 to prevent P-SV cancer development suggests that soy meal contained a factor(s) that blocked the antiandrogenic action of the phytoestrogen. CONCLUSIONS: The spontaneous development of P-SV cancers was significantly prevented in L-W rats consuming the SPII diet from age 2-24 months, possibly through an agonist effect of the soy derived phytoestrogens. PMID- 11027409 TI - Replication linkage study for prostate cancer susceptibility genes. AB - BACKGROUND: Since the publication of the first genome screen for prostate cancer (CaP) 5 years ago, over a dozen linkage studies have appeared. Most attention has been directed to chromosome 1, where two separate regions have been identified as harboring a prostate cancer susceptibility locus: HPC1 in the 1q24-25 interval and PCaP in the 1q42.2-43 interval. Linkage analysis of chromosome 16 has also provided evidence of harboring two loci predisposing to CaP. METHODS: We report on a replication linkage study of chromosomes 1 and 16 in 45 new and 4 expanded multiplex CaP families. Multipoint Z-scores were obtained for 30 highly polymorphic short-sequence tandem repeat markers spanning chromosome 1, and 22 markers spanning chromosome 16. RESULTS: The replication sample gave no evidence for a CaP susceptibility locus in the 1q24-25 interval and equivocal evidence for such a locus at 1q42.2-43. With respect to chromosome 16, positive Z-scores were obtained over a contiguous interval covering the entire p arm and the proximal half of the q arm. CONCLUSIONS: The linkage analysis of our replication sample does not support the existence of HPC1, and the evidence for the existence of PCaP remains equivocal. Evidence of a susceptibility locus on 16p remains strong, but the evidence for a susceptibility locus on 16q is weakened. PMID- 11027410 TI - Cultured stromal cells: an in vitro model of prostatic mesenchymal biology. AB - BACKGROUND: Initial efforts to develop in vitro models to study prostatic biology focused on the culture and characterization of epithelial cells. Recently, attention has turned towards inclusion of stromal cells in experimental systems. METHODS: Improved methods to isolate and culture stromal cells have been developed. An array of markers are employed to characterize subtypes of stromal cells, with particular interest in smooth muscle differentiation. RESULTS: Defined, serum-free media are available for certain experimental applications. Conditions that promote smooth muscle differentiation have been identified. Investigators have characterized hormonal and peptide factors that regulate the growth of prostatic stromal cells, and have also described paracrine factors produced by stromal cells that influence epithelial biology. CONCLUSIONS: Prostatic stromal-cell cultures are now widely employed by a large number of investigators for a diverse array of experimental purposes. While further refinement is required to obtain model systems that fully mimic in vivo processes, the availability of stromal- and epithelial-cell cultures provides a valuable resource for studying normal prostatic biology as well as benign prostatic hyperplasia (BPH) and cancer. PMID- 11027411 TI - Expression of androgen receptor coregulatory proteins in prostate cancer and stromal-cell culture models. AB - BACKGROUND: Androgen receptor (AR) transcriptional activity is modulated by cofactor proteins. They act as costimulators, corepressors, or bridging proteins, and a disbalanced expression may contribute to the altered activity of the AR in advanced prostate cancer. We investigated the expression of a series of steroid receptor cofactors in prostate cancer cell lines, including several LNCaP sublines, and in prostate stromal cells. METHODS: Expression of cofactors was analyzed by means of RT-PCR in PC-3, Du-145, LNCaP, three sublines of LNCaP established after long-term androgen deprivation, and two strains of primary prostate stroma cells. Expression in LNCaP and LNCaP-abl cells (which represented an advanced tumor cell) was analyzed employing semiquantitative RT-PCR. RESULTS: Ten of the 12 cofactors tested were expressed in all cells analyzed (AIB1, ARA54, ARA70, CBP, cyclin D1, Her2/neu/erbB2, BAG-1/M/L, SRC-1, SMRT, and TIF2). Only ARA55 and FHL2 mRNAs were not detected in all cells. ARA55 mRNA was absent in LNCaP cells, LNCaP sublines, and DU-145 cells; FHL2 was not expressed in LNCaP cells and its derivatives. The expression pattern was identical in LNCaP cells, and the long-term androgen ablated LNCaP sublines. Moreover, comparison of expression levels in LNCaP and LNCaP-abl cells revealed a slight reduction in LNCaP-abl cells but no gross differences. CONCLUSIONS: Prostatic cells express a great number of steroid receptor cofactors. AR activity thus seems to be modulated in a very complex way in prostate cells. PMID- 11027412 TI - Enzymatic action of prostate-specific antigen (PSA or hK3): substrate specificity and regulation by Zn(2+), a tight-binding inhibitor. AB - BACKGROUND: In semen, prostate-specific antigen (PSA or hK3) digests the gel proteins semenogelin I and II, resulting in liquefaction and the release of motile spermatozoa. We characterized the substrate specificity and zinc-mediated inhibition of PSA. METHODS: The proteolysis of human semenogelin I (SgI) and II (SgII) by PSA was characterized by purification of generated SgI and SgII fragments, N-terminal sequencing, and mass spectrometry. Zn(2+)-inhibition of PSA was studied using a chromogenic substrate. RESULTS: Eighteen cleavage sites in SgI and 16 in SgII were identified. Cleavages were identified mainly as the C terminal of certain tyrosine and glutamine residues, but also the C-terminal of histidine, aspartic acid, leucine, serine, and asparagine residues. No cleavages were identified at any arginine, lysine, phenylalanine, tryptophan, or methionine residues, indicating that the substrate specificity of PSA is distinct from that of trypsin, chymotrypsin, tissue kallkrein (hK1), and kallikrein 2 (hK2). Zn(2+) ions have a dramatic effect on PSA activity; the data indicate that Zn(2+) is a tight-binding inhibitor of PSA activity. CONCLUSIONS: The data will enable the optimized design of PSA activity assays, which may prove instrumental to uncovering the role of PSA in cancer and reproduction. The inhibition data indicate that Zn(2+) could regulate PSA activity, which may prove important in the development of efficient inhibitors of PSA activity. PMID- 11027413 TI - Rational basis for Trk inhibition therapy for prostate cancer. AB - BACKGROUND: Prostatic cancer cells are lethal because they acquire the ability to activate survival pathways that do not require androgenic stimulation. As a rational approach to developing effective therapy for these devastating cells, specific signal transduction pathways uniquely required for the survival of these nonandrogen-dependent prostate cancer cells must be identified. Previous studies suggested that the neurotrophin/trk signal transduction axis may regulate such unique survival pathways. In the present study, the changes in expression of the neurotrophins (NGF, BDNF, and NT-3) and their cognate receptors (i.e., trk and p75NTR) during the progression of normal prostatic epithelial cells to malignancy were documented. Additionally, the consequences of inhibiting these trk signaling pathways on the in vitro survival of prostate cancer cells was tested. METHODS: Immmunocytochemistry, RT-PCR, and ELISA assays were used to characterize the changes in the neurotrophin ligands (i.e., NGF, BDNF, and NT-3) and their cognate high-affinity (i.e., trk A, B, and C) and low-affinity neurotrophin (i.e., p75 NTR) receptors in normal vs. malignant human prostatic tissues. CEP-751 is an indolocarbazole compound specifically designed to inhibit the initiation of these neurotrophin/trk signal transductions. The consequence of CEP-751 inhibition of trk signaling for in vitro clonogenic survival of a series of human prostatic cancer lines was also tested. RESULTS: These studies demonstrated that normal prostatic tissue from patients without prostate cancer contains substantial levels of nerve growth factor (NGF), which is produced in a paracrine manner by stromal cells. These stromal cells lack both trk and p75NTR receptors. In contrast, normal prostatic epithelial cells from patients without prostate cancer do not secrete detectable levels of neurotrophins, but do express trk A and p75 NTR. While the NGF/trkA/p75 NTR axis is present in the normal prostate, normal prostatic epithelial cells do not depend on this axis for their survival. In contrast, malignant prostate epithelial cells directly secrete a series of neurotrophins (i.e., NGF, BDNF, and/or NT-3) and express at least one if not more of the trk receptor proteins (i.e., trk A, B, and/or C), while no longer expressing the p75NTR receptors. In addition, inhibition of autocrine trk signaling via CEP-751 treatment induces the apoptotic death of these malignant cells. CONCLUSIONS: Prostate carcinogenesis involves molecular changes leading to the paracrine and/or autocrine production of a series of neurotrophins. This is coupled to the ectopic expression of trk B and trk C, as well as to the continued expression of trk A, and the loss of expression of p75NTR receptors. These changes result in the acquisition by malignant prostate cells of a unique requirement for trk signaling pathways for survival. Based on these findings, trk inhibition is a novel, rational approach for prostate cancer therapy. PMID- 11027414 TI - Prostate-specific suicide gene therapy using the prostate-specific membrane antigen promoter and enhancer. AB - BACKGROUND: Prostate-specific membrane antigen (PSMA) is abundantly expressed in virtually 100% of prostate cancers and metastases. In addition, unlike prostate specific antigen (PSA), PSMA is upregulated under conditions of androgen deprivation. Therefore, PSMA is an attractive therapeutic target for advanced prostate cancer. Recently, both the promoter and the enhancer driving prostate specific expression of the PSMA gene were cloned. We describe here our analysis of the PSMA enhancer for the most active region(s) and present a way of using the enhancer in combination with the E. coli cytosine deaminase gene for suicide driven gene therapy that converts the nontoxic prodrug 5-fluorocytosine (5-FC) into the cytotoxic drug 5-fluorouracil (5-FU) in prostate cancer cells. METHODS: Deletion constructs of the full-length PSMA enhancer were subcloned into a luciferase reporter vector containing either the PSMA or SV-40 promoter. The most active portion of the enhancer was then determined via luciferase activity in the C4-2 cell line. We then replaced the luciferase gene with the E. coli cytosine deaminase gene in the subclone that showed the most luciferase activity. The specificity of this technique was examined in vitro, using the prostate cancer cell line LNCaP, its androgen-independent derivative C4-2, and a number of nonprostatic cell lines. The toxicity of 5-FC and 5-FU on transiently transfected cell lines was then compared. RESULTS: The enhancer region originally isolated from the PSMA gene was approximately 2 kb. Deletion constructs revealed that at least two distinct regions seem to contribute to expression of the gene in prostate cancer cells, and therefore the best construct for prostate-specific expression was determined to be 1, 648 bp long. The IC(50) of 5-FC was similar in all cell lines tested (>10 mM). However, transfection with the 1648 nt PSMA enhancer and the PSMA promoter to drive the cytosine deaminase gene enhanced toxicity in a dose-dependent manner more than 50-fold, while cells that did not express the PSMA gene were not significantly sensitized by transfection. CONCLUSIONS: Suicide gene therapy using the PSMA enhancer may be of benefit to patients who have undergone androgen ablation therapy and are suffering a relapse of disease. PMID- 11027415 TI - Peptide analogs in the therapy of prostate cancer. AB - The use of peptide analogs in the therapy of prostate cancer is reviewed. The preferred primary treatment of advanced androgen-dependent prostate cancer is presently based on the use of depot preparations of LH-RH agonists. This treatment is likewise recommended in patients with rising PSA levels after surgery or radiotherapy. LH-RH agonists with or without antiandrogens can be also utilized prior to or following various local treatments in patients with clinically localized prostate cancer and at high risk for disease recurrence. LH RH antagonists like Cetrorelix are in clinical trials. However, most patients with advanced prostatic carcinoma treated by any modality of androgen deprivation eventually relapse. Treatment of relapsed androgen-independent prostate cancer remains a major challenge, but new therapeutic modalities are being developed based on antagonists of growth hormone-releasing hormone (GH-RH) and bombesin, which inhibit growth factors or their receptors. Another approach consists of cytotoxic analogs of LH-RH, bombesin, and somatostatin containing doxorubicin or 2-pyrrolinodoxorubicin, which can be targeted to receptors for these peptides found in prostate cancers and their metastases. These cytotoxic analogs inhibit growth of experimental androgen-dependent or -independent prostate cancers and reduce the incidence of metastases. A rational therapy with peptide analogs could be selected on the basis of receptors present in biopsy samples. The approaches based on peptide analogs should result in a more effective treatment for prostate cancer. PMID- 11027416 TI - TGF-beta-based immunotherapy for cancer: breaching the tumor firewall. AB - Many malignant cells secrete transforming growth factor-beta (TGF-beta), a potent immunosuppresant, suggesting that TGF-beta production may represent a significant tumor escape mechanism from host immunosurveillance. Establishment of a leukocyte subpopulation with disrupted TGF-beta signaling in the tumor-bearing host offers a potential means for immunotherapy of cancer. Downregulation of TGF-beta secretion in tumor cells results in restoration of immunogenicity in the host, while T-cell insensitivity to TGF-beta results in accelerated differentiation and autoimmunity, elements of which may be required in order to combat self-antigen expressing tumors in a tolerized host. The rationale, approaches, and potential pitfalls of this strategy will be discussed. PMID- 11027417 TI - Clinical and pathologic aspects of spontaneous canine prostate carcinoma: a retrospective analysis of 76 cases. AB - BACKGROUND: Pet dogs and men share a vulnerability for the development of prostate carcinoma. The purpose of this study was to further characterize the clinical and pathologic features of spontaneous canine prostate carcinoma. METHODS: A multiinstitutional, retrospective study was conducted using 76 dogs with prostate carcinoma that underwent postmortem evaluation. For each case, clinical and pathologic data were tabulated and hematoxylin/eosin-stained tissue sections from the primary tumor and metastatic lesions were evaluated. Prostatic carcinomas were subclassified based upon the presence of glandular, urothelial, squamoid, or sarcomatoid differentiation. We focused our analysis on dogs that differed with respect to morphologic features of the primary tumor, lifetime duration of testicular hormone exposure, and presence of skeletal metastases. RESULTS: The vast majority of canine prostate carcinomas affected elderly sexually intact dogs or dogs that underwent surgical castration after sexual maturity. Adenocarcinoma was the most frequent histologic type, although more than half of canine prostate carcinomas exhibited intratumoral heterogeneity. In many cases, primary tumors showed mixed morphology, characterized by two or more types of differentiation. Duration of testicular hormone exposure was significantly different between dogs with adenocarcinoma and dogs with mixed morphology tumor, but did not appear to influence the frequency or pattern of metastases. Overall, gross metastases were present in 80% of dogs with prostate carcinoma. Skeletal metastases were present in 22% of cases, and the predominantly axial skeletal distribution of these lesions was similar to that reported in men with prostate carcinoma. Young dogs were at highest risk for development of skeletal metastases. CONCLUSIONS: This study provides a more complete characterization of spontaneous prostate carcinoma of dogs in terms of morphologic heterogeneity, skeletal metastases, and the influence of testicular hormones. Prostate carcinoma in pet dogs provides an immunocompetent, autochthonous tumor system that mimics certain aspects of human prostate cancer. This spontaneous model may contribute to our understanding of the factors that regulate carcinogenesis within the aged prostate, and to the development of chemoprevention strategies or bone-targeted therapies. PMID- 11027418 TI - Chemotherapy for hormone-refractory prostate cancer: beauty is in the eye of the beholder. PMID- 11027419 TI - International Consultation on Urological Diseases: a decade of progress. AB - The International Consultation on Urological Diseases (ICUD), under the cosponsorship of the World Health Organization (WHO), the International Society of Urology (SIU), and the International Union Against Cancer (UICC), is active in the organization of global consultations on urological diseases. The major urological associations from the five continents (the American Urological Association (AUA), the Urological Association of Asia (UAA), the Confederation Americana Urologia (CAU), and the European Association of Urology (EAU)), together with societies with expertise in specific programs such as the International Continence Society, the International Society for Impotence Research, the International Prostate Health Council, the American Cancer Society, the International Union Against Cancer, and the European Organization for Research and Treatment of Cancer, came together in the late 1980s on the basis of their proven record in organizing successful international meetings on urological cancer to initiate the concept of the ICUD. At the suggestion of the WHO, a First International Consultation on Benign Prostatic Hypertrophy (BPH) was organized in 1991, not only to provide recommendations to medical practitioners but also to define and plan future research on the subject. The formula, based on global multiprofessional collaboration and expertise, exceeded expectations, and other consultations were to follow, e.g., on prostate cancer, incontinence, erectile dysfunctions, and nosocomial infections. In 1994 it became necessary to establish a legal nongovernmental organization to foster the goals of the ICUD, essentially for the creation of awareness, knowledge, and recommendations on the diagnosis and treatment of urological diseases. PMID- 11027420 TI - Ciclopirox nail lacquer topical solution 8%. AB - Ciclopirox nail lacquer 8% (Penlac, Aventis Pharma) was approved by the US FDA in December 1999, as a component of a comprehensive management program, for use in immunocompetent patients who have mild to moderate onychomycosis of the fingers and toes without lunula involvement due to Trichophyton rubrum. The comprehensive management program includes removal of the unattached, infected nails as frequently as once per month, by a health care professional who has special competence in the diagnosis and treatment of nail disorders, including minor nail procedures. The nail lacquer is not approved in Canada. PMID- 11027421 TI - Treatment options for the cutaneous manifestations of systemic sclerosis. AB - Systemic Sclerosis is a multisystem disorder with vascular instability as a clinical hallmark. Treatment currently consists of recognition and management of end-organ damage. Dermatologists can assist in the management of these patients by facilitating early diagnosis, and treating cutaneous manifestations such as Raynaud's phenomenon, cutaneous calcinosis, and digital ulceration. New potentially disease-modifying therapies are now undergoing clinical trials. PMID- 11027422 TI - Is there a role for retinoids to treat minimal residual disease in neuroblastoma? AB - A variety of pre-clinical and clinical data point toward high drug levels of retinoids being required to achieve optimal efficacy against neuroblastoma. The results of the Kohler trial reported in this issue demonstrate that low-dose 13 cis-RA does not have clinical efficacy against neuroblastoma in a setting of minimal residual disease. A comparison of the Kohler trial with the US CCG trial provides clinical evidence that high-dose levels of retinoids are optimal for treating minimal residual disease in neuroblastoma. The comparison of high-dose and low-dose 13-cis-RA studies in neuroblastoma suggests the intriguing possibility that high dose, pulse schedules of other retinoids could be effective as therapeutic and chemopreventive agents in diseases where low-dose, chronic retinoid administration was not effective. Pre-clinical and perhaps clinical studies of the latter concept should be considered. PMID- 11027423 TI - A randomized trial of 13-Cis retinoic acid in children with advanced neuroblastoma after high-dose therapy. AB - One hundred and seventy-five children with Stage 3 or 4 neuroblastoma who had obtained a good response to conventional therapy were randomly allocated to 13 Cis retinoic acid at a dose of 0.75 mg/kg/day or placebo for up to 4 years. Toxicity was mild but no advantage in event-free survival was shown for the children receiving retinoic acid. PMID- 11027424 TI - Phase III randomized trial comparing moderate-dose cisplatin to combined cisplatin and carboplatin in addition to mitomycin and ifosfamide in patients with stage IV non-small-cell lung cancer. AB - A phase III randomized trial was conducted in patients with metastatic NSCLC, to determine if, in association with mitomycin (6 mg m(-2)) and ifosfamide (3 g m( 2)), the combination of moderate dosages of cisplatin (60 mg m(-2)) and carboplatin (200 mg m(-2)) - CarboMIP regimen - improved survival in comparison with cisplatin (50 mg m(-2)) alone - MIP regimen. A total of 305 patients with no prior chemotherapy were randomized, including 297 patients assessable for survival (147 in the MIP arm and 150 in the CarboMIP arm) and 268 patients assessable for response to chemotherapy. All but eight (with malignant pleural effusion) had stage IV disease. There was a 27% (95% CI, 19-34) objective response (OR) rate to MIP (25% of the eligible patients) and a 33% (95% CI, 24 41) OR rate to CarboMIP (29% of the eligible patients). This difference was not statistically significant (P = 0.34). Duration of response was not significantly different between both arms. There was also no difference (P = 0.67) in survival: median survival times were 28 weeks (95% Cl, 24-32) for MIP and 32 weeks (95% Cl, 26-35) for CarboMIP, with respectively 1-year survival rates of 24% and 23% and 2 year survival rates of 5% and 2%. The main toxicities consisted in emesis, alopecia, leucopenia and thrombocytopenia, that were, except alopecia, significantly more severe in the CarboMIP arm. Our trial failed to demonstrate a significant improvement in response or survival when patients with metastatic NSCLC were treated, in addition to ifosfamide and mitomycin, by combination of moderate dosages of cisplatin and carboplatin instead of moderate dosage of cisplatin alone. The results support the use of a moderate dose (50 mg m(-2)) of cisplatin in combination with ifosfamide and mitomycin for the chemotherapy of this disease. PMID- 11027425 TI - Chromosome instability in fibroblasts derived from Li-Fraumeni syndrome families without TP53 mutations. AB - The mean in vitro lifespan of dermal fibroblast strains derived from cancer affected individuals belonging to families conforming to the classical Li Fraumeni-syndrome or the Li-Fraumeni-like syndrome (LF strains), but in whom no TP53 mutation has been found, was not significantly different to that of normal strains. This was in contrast to LF strains that carry TP53 mutations. Cytogenetic observations of numerical and structural chromosome abnormalities were made on Giemsa stained metaphases prepared at different times during the lifespan of strains. Five strains from different LF families showed significantly increased frequencies of abnormal cells during the last 10% of their lifetime compared with seven normal strains and three other LF strains fell outside the normal range but did not reach significance. Two LF strains fell within the normal range indicating heterogeneity of the phenotype in this subset of LF fibroblasts. Numerical aberrations were the major aberration type observed. These observations of genetic instability are similar, but generally less strongly expressed, to those seen in LF strains with TP53 mutations. The basis for genetic instability in LF strains without TP53 mutations is not known, but appears not to involve defects in either the G(1)checkpoint or the checkpoint kinase hChk2. PMID- 11027426 TI - Preoperative serum CD26 levels: diagnostic efficiency and predictive value for colorectal cancer. AB - CD26 is an ectoenzyme with dipeptidyl peptidase IV activity expressed on a variety of cell types. Although the function of the high concentration of serum soluble CD26 (sCD26) is unknown, it may be related to the cleavage of biologically active polypeptides. As CD26 or enzymatic activity levels were previously associated with cancer, we examined the potential diagnostic and prognostic value of preoperative sCD26 measurements by ELISA in colorectal carcinoma patients. We found a highly significant difference between sCD26 levels in healthy donors (mean 559.7 +/- 125.5 microg l(-1)) and cancer patients (mean 261.7 +/- 138.1 microg l(-1)) (P< 0.001). A cut-off at 410 microg l(-1)gave 90% sensitivity with 90% specificity which means that the diagnostic efficiency of sCD26 is higher than that shown by other markers, particularly in patients at early stages. Moreover, sCD26 as a variable is not related with Dukes' stage classification, age, gender, tumour location or degree of differentiation. With a follow-up of 2 years until recurrence, preliminary data show that sCD26 can be managed as a prognostic variable of early carcinoma patients. In addition, the origin of sCD26 is discussed. PMID- 11027427 TI - Hepatocyte growth factor/scatter factor enhances the invasion of mesothelioma cell lines and the expression of matrix metalloproteinases. AB - Hepatocyte growth factor/scatter factor (HGF/SF) is a multifunctional factor involved both in development and tissue repair, as well as pathological processes such as cancer and metastasis. It has been identified in vivo in many types of tumours together with its tyrosine kinase receptor, Met. We show here that exogenous HGF/SF acts as a strong chemoattractant for human mesothelioma cell lines. The factor also enhanced cell adhesion to and invasion into Matrigel. The mesothelioma cell lines synthesized a panel of matrix metalloproteinases critical for tumour progression such as MMP-1, 2, 3, 9 and membrane-bound MT1-MMP. HGF/SF stimulated the expression of MMP-1, 9 and MT1-MMP and had a slight effect on expression of the MMP inhibitor TIMP-1 but not TIMP-2. However, there was no simple correlation between the levels of MMPs and TIMPs of the cell lines and their different invasion properties or between HGF/SF stimulatory effects on MMP expression and invasion. In addition, effects of protease inhibitors on invasion suggested that serine proteases were also expressed in human mesothelioma cell lines and were involved in HGF/SF-induced invasion. The results show a predominant role for HGF/SF in mesothelioma cell invasion, stimulating simultaneously adhesion, motility, invasion and regulation of MMP and TIMP levels. PMID- 11027428 TI - Expression and function of angiopoietin-1 in breast cancer. AB - Angiopoietin-1 (Ang1) has been shown to act as an angiogenic promoter in embryonic angiogenesis by promoting vascular branching, pericyte recruitment and endothelial survival. We have investigated the role of Ang1 in tumour neovascularization under clinical conditions and in animal models. The expression of Ang1 in clinical breast cancer specimens was analysed by using laser-capture microdissection and reverse transcriptase-linked polymerase chain reaction (RT PCR) on RNA isolated from the samples. Despite the expression of Ang1 in many human breast cancer cell lines, the gene was expressed in only three of 21 breast cancer clinical specimens, even though its receptor, Tie2, is abundant in the vasculature of all of these tumours. Ang1 was then overexpressed in a human breast cancer cell line (MCF-7) on its own and in conjunction with FGF1, an angiogenic factor shown to be able to increase the tumorigenicity of MCF-7 cells. High concentrations of Ang1 were produced in the conditioned media of the transfected cells (range 156-820 ng ml(-1)). However, in contrast to its physiological role as promoter of angiogenesis, overexpression of Ang1 did not enhance tumour growth, but instead caused up to a 3-fold retardation of tumour growth (P = 0.003). PMID- 11027429 TI - Cell cycle regulators p27 and pRb in lymphomas - correlation with histology and proliferative activity. AB - The cell cycle is a complex event in which multiple regulator-proteins participate. The G(1)/S checkpoint of the cell cycle is controlled by pRb protein, which functions in its hypophosphorylated form as a negative regulator of growth. p27 (Kip1), a member of CIP/KIP family of cyclin inhibitory proteins, participates in inhibition of forming complexes that allow pRb to phosphorylate and lead the cell into mitosis. The expression of these important cell cycle regulator proteins was studied in a total of 96 non-Hodgkin's lymphoma (NHL) samples, which were classified according to the REAL classification. The expression of p27, pRb and the cell proliferation marker Ki-67 (MIB-1) was evaluated in lymphomas using immunohistochemistry. This study showed that there were coordinate changes in the expression of p27 and pRb in NHL. When compared to low-grade lymphomas, high-grade lymphomas showed significantly reduced expression of p27 and inversely pRb expression was increased (P< 0.001). Increase in expression of Ki-67 was parallel with pRb expression, and was mainly seen in cells that lacked p27 expression (P< 0.0001). This study suggests that changes in the control of the cell cycle closely relate to the pathobiology of NHL. PMID- 11027430 TI - Changes in E-cadherin associated with cytoplasmic molecules in well and poorly differentiated endometrial cancer. AB - E-cadherin function is thought to be impaired in epithelial cancer. To investigate the alterations in E-cadherin associated with cytoplasmic molecules including alpha-catenin, beta-catenin, gamma-catenin, p120CAS, and IQGAP1 in various endometrial cancers with different degree of differentiation, we examined the localization and expression of E-cadherin and cytoplasmic molecules in 30 cases of both well and poorly differentiated endometrioid adenocarcinomas, using immunofluorescence and immunoblotting techniques. E-cadherin and cytoplasmic molecules demonstrated linear staining at the cell boundaries in normal endometrium. In all 20 cases with well differentiated adenocarcinomas, alpha catenin and IQGAP1 disappeared from the cell adhesive sites, but other cytoplasmic molecules were co-localized with E-cadherin along the cell boundaries. In all 10 cases with poorly differentiated adenocarcinomas, E cadherin and cytoplasmic molecules accumulated as large aggregates along cell adhesive sites, and the localization of IQGAP1 differed from those of other cytoplasmic molecules. The expression of these molecules in all 20 cases with well differentiated adenocarcinomas decreased or was lost in Triton-insoluble fraction, in comparison with the findings for all cases with normal endometrium or poorly differentiated adenocarcinomas. These results suggested that each alteration in E-cadherin associated with cytoplasmic molecules may play a different role in E-cadherin dysfunction between well and poorly differentiated adenocarcinomas. PMID- 11027431 TI - Nitric oxide synthase inhibition results in synergistic anti-tumour activity with melphalan and tumour necrosis factor alpha-based isolated limb perfusions. AB - Nitric oxide (NO) is an important molecule in regulating tumour blood flow and stimulating tumour angiogenesis. Inhibition of NO synthase by L-NAME might induce an anti-tumour effect by limiting nutrients and oxygen to reach tumour tissue or affecting vascular growth. The anti-tumour effect of L-NAME after systemic administration was studied in a renal subcapsular CC531 adenocarcinoma model in rats. Moreover, regional administration of L-NAME, in combination with TNF and melphalan, was studied in an isolated limb perfusion (ILP) model using BN175 soft tissue sarcomas. Systemic treatment with L-NAME inhibited growth of adenocarcinoma significantly but was accompanied by impaired renal function. In ILP, reduced tumour growth was observed when L-NAME was used alone. In combination with TNF or melphalan, L-NAME increased response rates significantly compared to perfusions without L-NAME (0-64% and 0-63% respectively). An additional anti-tumour effect was demonstrated when L-NAME was added to the synergistic combination of melphalan and TNF (responses increased from 70 to 100%). Inhibition of NO synthase reduces tumour growth both after systemic and regional (ILP) treatment. A synergistic anti-tumour effect of L-NAME is observed in combination with melphalan and/or TNF using ILP. These results indicate a possible role of L-NAME for the treatment of solid tumours in a systemic or regional setting. PMID- 11027432 TI - Loss of growth inhibitory effects of retinoic acid in human breast cancer cells following long-term exposure to retinoic acid. AB - Although retinoids are known to be inhibitory to breast cancer cell growth, a key remaining question is whether they would remain effective if administered long term. We describe here the long-term effects of all- trans retinoic acid on two oestrogen-dependent human breast cancer cell lines MCF7 and ZR-75-1. Although both cell lines were growth inhibited by retinoic acid in the short-term in either the absence or the presence of oestradiol, prolonged culture with 1 microM all- trans retinoic acid resulted in the cells acquiring resistance to the growth inhibitory effects of retinoic acid. Time courses showed that oestrogen deprivation of the cell lines resulted in upregulation of the basal non-oestrogen stimulated growth rate such that cells learned to grow at the same rate without as with oestradiol, but the cells remained growth inhibited by retinoic acid throughout. Addition of 1 microM all- trans retinoic acid to steroid deprivation conditions resulted in reproducible loss of growth response to both retinoic acid and oestradiol, although the time courses were separable in that loss of growth response to retinoic acid preceded that of oestradiol. Loss of growth response to retinoic acid did not involve loss of receptors, ER as measured by steroid binding assay or RARalpha as measured by Northern blotting. Function of the receptors was retained in terms of the ability of both oestradiol and retinoic acid to upregulate pS2 gene expression, but there was reduced ability to upregulate transiently transfected ERE- and RRE-linked reporter genes. Despite the accepted role of IGFBP3 in retinoic acid-mediated growth inhibition, progression to retinoic acid resistance occurred irrespective of level of IGFBP3, which remained high in the resistant MCF7 cells. Measurement of AP1 activity showed that the two cell lines had markedly different basal AP1 activities, but that progression to resistance was accompanied in both cases by a lost ability of retinoic acid to reduce AP1 activity. These results warn of potential resistance which could arise on long-term treatment with retinoic acid in a clinical situation and echo the problems of progression to endocrine resistance. It seems that whatever the constraints imposed on growth, these cells have a remarkable ability to escape from growth inhibition. However, the ability of retinoic acid to delay progression to oestrogen resistance is encouraging for endocrine therapy, and the concentration-dependence of retinoic acid resistance suggests that progression is not absolute but could be manipulated by dose. PMID- 11027433 TI - MHC class II antigen presentation pathway in murine tumours: tumour evasion from immunosurveillance? AB - Qualitative differences in the MHC class II antigen processing and presentation pathway may be instrumental in shaping the CD4+ T cell response directed against tumour cells. Efficient loading of many MHC class II alleles with peptides requires the assistance of H2-M, a heterodimeric MHC class II-like molecule. In contrast to the HLA-DM region in humans, the beta-chain locus is duplicated in mouse, with the H2-Mb1 (Mb1beta-chain distal to H2-Mb2 (Mb2) and the H2-Ma (Ma) alpha-chain gene). Here, we show that murine MHC class II and H2-M genes are coordinately regulated in murine tumour cell lines by T helper cell 1 (IFN-gamma) and T helper cell 2 (IL-4 or IL-10) cytokines in the presence of the MHC class II specific transactivator CIITA as determined by mRNA expression and Western blot analysis. Furthermore, Malphabeta1 and Malphabeta2 heterodimers are differentially expressed in murine tumour cell lines of different histology. Both H2-M isoforms promote equally processing and presentation of native protein antigens to H2-A(d)- and H2-E(d)-restricted CD4+ T cells. Murine tumour cell lines could be divided into three groups: constitutive MHC class II and CIITA expression; inducible MHC class II and CIITA expression upon IFN-gamma-treatment; and lack of constitutive and IFN-gamma-inducible MHC class II and CIITA expression. These differences may impact on CD4+ T cell recognition of cancer cells in murine tumour models. PMID- 11027434 TI - A transgenic mouse model for tumour immunotherapy: induction of an anti-idiotype response to human MUC1. AB - MUC1 is a membrane bound, polymorphic epithelial mucin expressed at the luminal surface of glandular epithelium. It is highly expressed in an underglycosylated form on carcinomas and metastatic lesions and is, therefore, a potential target for immunotherapy of cancer. The monoclonal antibody HMFG1 binds the linear core protein sequence, PDTR, contained within the immunodominant domain of the tandem repeat of MUC1. The efficacy of murine and humanized HMFG1 (Ab1) used as an anti idiotypic vaccine was examined in mice transgenic for human MUC1 (MUC1.Tg) challenged with murine epithelial tumour cells transfected with human MUC1. Humoral idiotypic cascade through Ab2 and Ab3 antibodies was observed in MUC1.Tg mice following multiple antibody inoculations in the presence of adjuvant. Impaired tumour growth at day 35 and highest Ab3 levels were found in mice that had received mHMFG1 with RAS adjuvant. However, comparison of Ab3 levels in individual mice with tumour size in all treatment groups did not show a correlation between smaller tumours and increased levels of anti-idiotype antibody. This suggests that the anti-tumour effects of anti-idiotype vaccination are not solely related to the induction of idiotypic antibody cascades and probably involve other mechanisms. PMID- 11027436 TI - Overexpression of melanoma inhibitory activity (MIA) enhances extravasation and metastasis of A-mel 3 melanoma cells in vivo. AB - The secreted MIA protein is strongly expressed by advanced primary and metastatic melanomas but not in normal melanocytes. Previous studies have shown that MIA serum levels correlate with clinical tumour progression in melanoma patients. To provide direct evidence that MIA plays a role in metastasis of malignant melanomas, A-mel 3 hamster melanoma cells were transfected with sense- and antisense rhMIA cDNA and analysed subsequently for changes in their tumorigenic and metastatic potential. Enforced expression of MIA in A-mel 3 cells significantly increased their metastatic potential without affecting primary tumour growth, cell proliferation or apoptosis rate in hamsters, compared with control or antisense transfected cells. Additionally, MIA overexpressing transfectants showed a higher rate of both tumour cell invasion and extravasation. Cells transfected with MIA antisense generally exerted an opposite response. The above changes in function attributed to the expression of MIA may underlie the contribution of MIA to the malignant phenotype. PMID- 11027435 TI - Downregulation of intracellular nm23-H1 prevents cisplatin-induced DNA damage in oesophageal cancer cells: possible association with Na(+), K(+)-ATPase. AB - Previously, we showed that expression of nm23-H1 is associated inversely with sensitivity to cisplatin in human oesophageal squamous cell carcinoma (OSCC). The present study was undertaken to investigate the association of nm23-H1 expression with cisplatin-induced DNA damage in OSCC using antisense nm23-H1 transfectants. YES-2/AS-12, an antisense nm23-H1-transfected OSCC cell line, showed significantly reduced expression of intracellular nm23-H1 protein compared with that in parental YES-2 cells and YES-2/Neo transfectants. Surface expression of nm23-H1 protein was not observed in any of the three cell lines. PCR analysis for DNA damage demonstrated that YES-2/AS-12 cells were more resistant to nuclear and mitochondrial DNA damage by cisplatin than were YES-2/Neo cells. In addition, mitochondrial membrane potentials and DNA fragmentation assays confirmed that YES 2/AS-12 was more resistant than YES-2/Neo to apoptosis induced by cisplatin. In contrast, YES-2/AS-12 was more sensitive to ouabain, a selective inhibitor of Na(+), K(+)-ATPase, than YES-2 and YES-2/Neo. Pre-treatment with ouabain resulted in no differences in cisplatin sensitivity between the three cell lines examined. Intracellular platinum level in YES-2/AS-12 was significantly lower than that in YES-2 and YES-2/Neo following incubation with cisplatin, whereas ouabain pre treatment resulted in no differences in intracellular platinum accumulations between the three cell lines. Our data support the conclusion that reduced expression of intracellular nm23-H1 in OSCC cells is associated with cisplatin resistance via the prevention of both nuclear and mitochondrial DNA damage and suggest that it may be related to Na(+), K(+)-ATPase activity, which is responsible for intracellular cisplatin accumulation. PMID- 11027437 TI - Production of a signal by irradiated cells which leads to a response in unirradiated cells characteristic of initiation of apoptosis. AB - This study investigated the ability of medium from irradiated cells to induce early events in the apoptotic cascade, such as mobilization of intracellular calcium, loss of mitochondrial membrane potential and increase in reactive oxygen species, in cells which were never exposed to radiation. Medium from irradiated human keratinocytes was harvested and transferred to unirradiated keratinocytes. Endpoints characteristic of the initiation of apoptosis were monitored for a period of 24 h following medium transfer. Clonogenic survival was also measured. Rapid calcium fluxes (within 30 s), loss of mitochondrial membrane potential, increases in reactive oxygen species (from 6 h after medium transfer), an increase in the number of apoptotic cells (48 hours after medium transfer) and a marked reduction in clonogenic survival (after 9 days) were observed. There was no significant difference between medium generated by cells irradiated at 0.5 Gy or 5 Gy. The data suggest that initiating events in the apoptotic cascade were induced in unexposed cells by a signal produced by irradiated cells. PMID- 11027439 TI - Endocrine correlates of male breast cancer risk: a case-control study in Athens, Greece. AB - We studied the relation of certain endocrine-related variables among 23 cases of male breast cancer and 76 apparently healthy male controls. There were significant inverse associations with smoking (P = 0.03), birth order (P = 0.02) and reported frequency of orgasms in later life (P = 0.0004). The study provides strong indirect evidence that endocrine factors are important in the aetiology of male breast cancer. PMID- 11027438 TI - Breast cancer risk in male twins: joint analyses of four twin cohorts in Denmark, Finland, Sweden and the United States. AB - To test the hypothesis that in utero exposure to high levels of oestrogen increases the risk of male breast cancer, we followed 115 235 male twins for more than 3.5 million person-years at risk. We observed 11 cases of male breast cancer versus 16.16 expected based on national rates (standardized rate ratio 0.68, 95% confidence interval 0.34-1.22) and conclude that any adverse influence of in utero oestrogen exposure is likely to be small. PMID- 11027440 TI - Oral hygiene, dentition, sexual habits and risk of oral cancer. AB - In an Italian case-control study of oral cancer, number of missing teeth and other aspects of dental care were similar, but the general condition of the mouth, as indicated by gum bleeding, tartar deposits and mucosal irritation, was worse among oral cancer cases than controls. No differences were detected in sexual practices (including oral sex) and (previous) sexually transmitted infections. PMID- 11027441 TI - Sunscreen use and intentional exposure to ultraviolet A and B radiation: a double blind randomized trial using personal dosimeters. AB - A previous randomized trial found that sunscreen use could extend intentional sun exposure, thereby possibly increasing the risk of cutaneous melanoma. In a similarly designed trial, we examined the effect of the use of sunscreens having different sun protection factor (SPF) on actual exposure to ultraviolet B (UVB) and ultraviolet A (UVA) radiation. In June 1998, 58 European participants 18-24 years old were randomized to receive a SPF 10 or 30 sunscreens and were asked to complete daily records of their sun exposure during their summer holidays of whom 44 utilized a personal UVA and UVB dosimeter in a standard way during their sunbathing sessions. The median daily sunbathing duration was 2.4 hours in the SPF 10 group and 3.0 hours in the SPF 30 group (P = 0.054). The increase in daily sunbathing duration was paralleled by an increase in daily UVB exposure, but not by changes in UVA or UVB accumulated over all sunbathing sessions, or in daily UVA exposure. Of all participants, those who used the SPF 30 sunscreen and had no sunburn spent the highest number of hours in sunbathing activities. Differences between the two SPF groups in total number of sunbathing hours, daily sunbathing duration, and daily UVB exposure were largest among participants without sunburn during holidays. Among those with sunburn, the differences between the two groups tended to reduce. In conclusion, sunscreens used during sunbathing tended to increase the duration of exposures to doses of ultraviolet radiation below the sunburn threshold. PMID- 11027443 TI - Maternal pregnancy hormone profiles in areas with a different incidence of breast cancer. PMID- 11027442 TI - Oesophageal cancer among the Turkomans of northeast Iran. AB - A Caspian Littoral Cancer Registry survey in the early 1970s established northern Iran as one of the highest oesophageal cancer incidence regions of the world. To verify this, an oesophageal cancer survey was carried out between 1995 and 1997 in the Turkoman Plain at the southeastern corner of the Caspian Sea. Oesophageal balloon cytology screening was carried out on 4192 asymptomatic adults above age 30 years in one town and three adjoining villages with a total population of 20 392 people at risk. Oesophagoscopy was performed on 183 patients with abnormal cytological findings. The discovery of two asymptomatic small squamous cell cancers and one 'carcinoma- suspect' implied a prevalence ranging from 47.7 per 100 000 to 71.5 per 100 000. During a 1-year active surveillance, 14 patients were found with clinically advanced oesophageal squamous cell cancer, yielding age-standardized incidence rates of 144.09 per 100 000 for men and 48.82 per 100 000 for women. The very high frequency of oesophageal cancer reported for northern Iran 25 years ago stands confirmed. Differences in incidence rates, then and now, can be attributed to survey methods used and diagnostic criteria applied, but not to socioeconomic factors, which have remained relatively stable. Oesophageal balloon cytology is a practical method of mass screening for oesophageal cancer in Iran. PMID- 11027444 TI - Maternal pregnancy hormone profiles in areas with a different incidence of breast cancer - reply PMID- 11027445 TI - Serum lactate dehydrogenase isoenzyme 1 as a prognostic predictor for metastatic testicular germ cell tumours. PMID- 11027446 TI - Serum lactate dehydrogenase isoenzyme 1 as a prognostic predictor for metastatic testicular germ cell tumours - reply PMID- 11027447 TI - Galectins: a new family of regulators of inflammation. PMID- 11027448 TI - Immunization with the C-terminal region of Trypanosoma cruzi ribosomal P1 and P2 proteins induces long-term duration cross-reactive antibodies with heart functional and structural alterations in young and aged mice. AB - The R13 peptide sequence (EEEDDDMGFGLFD) that corresponds to the C-terminal region of Trypanosoma cruzi ribosomal P1 and P2 proteins differs from the eukariotic P concensus sequence EESDDDMGFGLFD (H13) only in a nonconservative amino acid substitution. The immunization of BALB/c mice with R13 synthetic peptide coupled to a carrier protein (OVA) induces specific (anti-R13) and autoreactive (anti-H13 and anti-heart) antibodies as well as heart functional alterations. Since aged human and experimental animals are impaired in their responses to most foreign antigens but they produce greater amounts of autoantibodies, in this work we used aged mice as an experimental model able to exaggerate the autoimmune component of the R13-induced response in case it was present. We studied whether these antibodies generated in the absence of the parasite would induce pathological changes in heart tissues. The levels of antibodies against R13 (foreign antigen) and H13 (autoantigen) studied comparatively in 2- and 12-month-old mice 10 days after the third immunization with R13 coupled to OVA were, as we expected for a foreign antigen, higher in almost all sera from 2-month-old mice tested than in sera from 12-month-old mice. Besides, these specific and cross-reactive antibody response remain elevated as long as 150 days post third immunization. In addition, the isotype pattern that recognizes R13 and the self-sequence H13 showed no differences between sera from young and aged mice. Moreover, when ECG traces were obtained from immunized mice, the heart functional alterations observed at 10 days continued at 80 and 150 days after the third immunization, showing an association with the levels of antibodies. In addition, despite the fact that the heart tissue morphology showed no alterations 10 days post third immunization, several abnormalities in the tissue architecture were revealed at 80 and 150 days post third immunization. This report demonstrates the biological relevance of R13-induced cross-reactive antibodies in some of the electrophysiologic and histological changes found in T. cruzi-infected mammalians. PMID- 11027449 TI - Evaluation of thymopoiesis using T cell receptor excision circles (TRECs): differential correlation between adult and pediatric TRECs and naive phenotypes. AB - To determine whether the thymus is still functional despite age-related involution, we measured a biomarker for thymopoiesis known as the T cell receptor excision circle (TREC) from peripheral blood mononuclear cells (PBMCs) of 148 healthy children and from PBMCs, CD4(+), and CD8(+) cells of 32, 30, and 50 healthy adults, respectively. We demonstrate that during the first 5 years of life, thymic output is decreased (P 0.002) but not dramatically (r = -0. 282). Among adults aged 23-58, thymic output was inversely correlated with age, as measured from PBMCs (r = -0.628, P < 0.0005), CD4(+) (r = -0.530, P 0.003), and CD8(+) fractions (r = -0.385, P 0. 006). A strong correlation existed between pediatric PBMC TRECs and the expression of three naive phenotypic markers (CD45RA(+)CD45RO(-), CD45RA(+)CD62L(+), and CD45RO(-)CD27(+)CD95(low)). Adult PBMC TRECs correlated only with the expression of CD45RA(+)CD45RO(-) (r = 0.459, P 0.012). Our data suggest that in adults CD45RA(+)CD45RO(-) may be enriched for TRECs and add to a growing body of evidence illustrating intact thymic function in adulthood. PMID- 11027450 TI - Ex vivo modulation of RANTES and sCD30 by proinflammatory stimuli in HIV seropositive and -negative individuals. AB - RANTES and sCD30 were measured in ex vivo culture supernatants of unstimulated or stimulated PBMC in order to investigate their potential role as markers of acute immune activation. Patients in an advanced stage of HIV infection (AIDS A) were compared to AIDS patients who were evaluated for pneumonia at the time of blood withdrawal (AIDS B); HIV(+) individuals with nonprogressive infection (LTNP) and healthy donors (N) served as controls. Constitutive levels of RANTES were significantly elevated in AIDS B patients (P 0.0001), whereas spontaneous release of sCD30 was strongly correlated with the presence of both pneumonia (P 0.002) and HIV infection (P 0.004). LPS was a strong inducer of RANTES in all four categories; however, in AIDS B patients a negative and positive correlation between constitutive and induced levels was observed with LPS (P 0.0004) and IFN gamma (P 0.006), respectively. We clearly showed that IFN-gamma reached a fourfold superinduction of sCD30 release in both HIV-positive and -negative individuals, whereas IL-6-driven production of both sCD30 and RANTES occurred only in healthy donors. Ex vivo RANTES levels may also be monitored as an index of acute immune activation under conditions of chronic activation of the immune system, whereas sCD30 release may be equally indicative of both acute and chronic processes of T cell activation. Proinflammatory stimuli differentially affected RANTES and sCD30 secretion in ex vivo PBMC cultures, suggesting complex pathways in the in vivo regulation of these two molecules. PMID- 11027452 TI - Bacteremia and skin/bone infections in two patients with X-linked agammaglobulinemia caused by an unusual organism related to Flexispira/Helicobacter species. AB - Two patients with Bruton's X-linked agammaglobulinemia are described with bacteremia and skin/bone infection due to an organism which by 16S rRNA gene sequence analysis was most closely related to "Flexispira" rappini (and thus designated a Flexispira-like organism, FLO) and more distantly related to the Helicobacter species. The organism required microaerobic conditions and, supplemental H(2) gas for growth and was reliably stained with acridine orange. In common with Helicobacter cinaedi infections, the focus of the FLO infection was in one case in the blood vessels or lymphatics of an extremity and in the other case in the skin and adjacent bone of an extremity. In both cases, prolonged IV antibiotic therapy was necessary to clear the infection. The susceptibility of XLA patients to FLO infection appears to be related to the fact that XLA is associated with severe B cell (humoral) immunodeficiency and thus these patients have difficulty with intravascular or intralymphatic infection. These findings elucidate the nature of FLO infections in humans and point the way to their detection and treatment. PMID- 11027451 TI - Immune stimulation in scleroderma patients treated with thalidomide. AB - Scleroderma (SSc) is a fibrosing connective tissue disease that is poorly responsive to any treatment, including immune suppression. SSc shares many characteristics with chronic graft-versus-host disease (GVHD). Because the immunomodulatory drug thalidomide has proven beneficial in chronic GVHD, we studied the immune response and clinical effects of thalidomide in SSc patients. We treated 11 SSc patients with thalidomide in an open label, dose escalating, 12 week study. Histologic comparison of skin biopsies showed changes in skin fibrosis and an increase in epidermal and dermal infiltrating CD8(+) T cells with thalidomide treatment. In thalidomide-treated SSc patients, plasma levels of IL 12 and TNF-alpha increased, while plasma IL-5 and IL-10 levels remained unchanged. These changes were associated with clinical effects, including dry skin, dermal edema, transient rashes, decreased gastroesophageal reflux symptoms, and healing of digital ulcers. When SSc PBMCs activated by anti-CD3 mAb were exposed to thalidomide, increases in both production of IL-2, IL-3, GM-CSF, and IFN-gamma and T cell expression of CD40L were observed. Thalidomide therefore appears to induce immune stimulation in SSc patients in association with clinical changes. However, it remains to be shown whether long-term enhancement of immune responses in SSc patients is clinically beneficial. PMID- 11027453 TI - Enhanced immunological tolerance against allograft rejection by oral administration of allogeneic antigen linked to cholera toxin B subunit. AB - A single oral intragastric administration of cholera toxin B subunit (CTB) conjugated to allogeneic thymocytes (ATC, 4 x 10(7) cells) under conditions allowing the CTB to bind the complex to GM1 ganglioside receptors was shown to be efficacious in inducing peripheral T cell tolerance associated with significant suppression of both primary and secondary accelerated rejection of heart allografts when tested in mice. Allogeneic in vivo delayed-type hypersensitivity (DTH), in vitro cytotoxicity responses, and mixed lymphocyte reactions (MLR) by T cells from mesenteric lymph nodes (MLN), popliteal lymph nodes (PLN), and spleen were significantly reduced in mice treated with the CTB-ATC conjugate, as were also the numbers of cells in these organs producing IL-2, IFN-gamma, or IL-4. In contrast, a marked increase in the production of IL-4 in Peyer's patches (PP) and of TGF-beta(1) in PLN was observed. The suppressive potential of T cells from PP and/or MLN after oral treatment with CTB-ATC was further evident by intraperitoneal transfer of such cells from CTB-ATC-treated animals to primed recipients, which led to marked suppression of both allogen-specific DTH and MLR responses. A critical role for PP in inducing peripheral tolerance after oral CTB ATC treatment was indicated by the absence of tolerance induction in animals whose PP had been destroyed before treatment with CTB-ATC. The results indicate that the protection against allograft rejection by oral treatment with CTB-ATC is mediated by T cells and associated with a strong induction of IL-4 production at mucosal sites and TGF-beta(1) at the effector sites. PMID- 11027454 TI - In vivo nasal challenge with diesel exhaust particles enhances expression of the CC chemokines rantes, MIP-1alpha, and MCP-3 in humans. AB - Diesel exhaust particles (DEP) enhance allergic inflammation by increasing in vivo IgE and cytokine production in the human upper respiratory mucosa. CC chemokines have been shown to play an important role in inflammation. We examined whether DEP could alter the production of CC chemokines by cells residing in the human nasal mucosa. At both 6 and 24 h following intranasal DEP challenge, the levels of nasal RANTES, MIP-1alpha, and MCP-3 were significantly elevated compared to baseline. In contrast, DEP did not enhance levels of Eotaxin at any time, demonstrating that the action of DEP was not simply a global effect on all CC chemokines. Challenge with saline resulted in no significant change in expression of any chemokine at any time. Challenge with DEP also resulted in an increase in total cell counts in nasal lavage fluids. Increases in lymphocyte, monocyte/macrophage, and neutrophil cells were observed but there was no change in eosinophil cell numbers. In contrast, there was a significant enhancement of ECP protein levels in washes performed 6 to 24 h after DEP challenge. Elevated specific nasal chemokine expression following exposure to DEP likely participates in the inflammation, cellular infiltration, and increase in IgE observed in the absence of allergen. PMID- 11027455 TI - Safety and immune responses to attenuated Salmonella enterica serovar typhi oral live vector vaccines expressing tetanus toxin fragment C. AB - Attenuated Salmonella enterica serovar Typhi vaccine strain CVD 908-htrA was used as a vector to deliver fragment C of tetanus toxin as a single-dose oral tetanus vaccine candidate to elicit protective levels of serum tetanus antitoxin. Twenty one healthy adult volunteers received doses of 1.6 x 10(7) to 8.2 x 10(9) CFU of one of two strains, CVD 908-htrA(pTETnir15) or CVD 908-htrA(pTETlpp), which contained plasmid-encoded fragment C, with sodium bicarbonate, and the safety and immune responses to serovar Typhi antigens and tetanus toxin were assessed. No volunteer had fever or positive blood cultures after vaccination, although diarrhea occurred in 3 volunteers and vomiting in 2 volunteers within 3 weeks after vaccination. Most volunteers excreted the vaccine strain in the first 72 h after vaccination. Three of nine volunteers who received 10(8) CFU or higher doses of the CVD 908-htrA(pTETlpp) construct developed rises in serum antitoxin antibodies. The serum and cellular immune responses to serovar Typhi antigens were less frequent than those previously observed in volunteers who ingested the parent strain CVD 908-htrA. This study demonstrates that fragment C of tetanus toxin delivered orally to volunteers in an S. Typhi vector can elicit protective levels of serum antitoxin. PMID- 11027456 TI - Immune response of HLA-DQ transgenic mice to house dust mite allergen p2: identification of HLA-DQ restricted minimal epitopes and critical residues. AB - HLA-DQ8 (HLA-DQA1*0301; HLA-DQB1*0302) and HLA-DQ6 (HLA-DQA1*0103; HLA-DQB1*0602) genes were introduced into mouse class II (H-2A(o)(beta)) knockout mice. Transgenic HLA-DQ8 and HLA-DQ6 mice were individually immunized and challenged using synthetic peptides representing HDM (Dermatophagoides pteronyssinus) allergen p2. HLA-DQ8 mice responded to p2 peptides 1-20, 41-60, 51-70, 61-80, 91 110, and 101-120. HLA-DQ6 mice responded to peptides 1-20, 11-30, 21-40, 41-60, and 51-70. Using single amino acid truncated 30-mer peptides, residues necessary for HLA-DQ8 recognition were identified spanning regions 3-12, 50-70, and 91-120. A synthetic peptide comprising residues 3-12 was synthesized and a series of single alanine substitutions was introduced into the minimal peptide. Introduction of alanine residues at positions 3, 11, and 12 resulted in a significant loss of immune recognition. It was concluded that residues 4, 5, 7, 11, and 12 are critical for immune recognition by HLA-DQ8 mice. PMID- 11027457 TI - Bulk cytokine production versus frequency of cytokine-producing cells in HIV1 infection before and during HAART. AB - Cytokine imbalances play a major role in HIV immunopathogenesis. This study analyzes simultaneously the frequency of cytokine-producing cells at the single cell level by flow cytometry and the disturbances in cytokine secretion assessed by ELISA in a cohort of asymptomatic HIV1 patients in different stages of CD4 depletion and during antiretroviral therapy (HAART). Early in the disease, there is an increased frequency of IFN-gamma(+) lymphocytes and bulk IFN-gamma production, in parallel with a reduced proportion of IL4(+) cells and IL4 secreted. The two IL4 measurements are significantly correlated. No such correlation was found for IFN-gamma, which is consistent with a large variation in the amount of IFN-gamma released per individual cell. Moreover, HAART was associated with a reduction to normal levels in the bulk IFN-gamma secretion concomitant with a persistency of the overexpanded IFN-gamma(+) cell subset in the peripheral blood. This study emphasizes the importance of using a conjoint approach to assess the cytokine network in trials of antiretroviral and/or immune based therapies to avoid missing significant effects which are possibly relevant in the clinical setting. PMID- 11027458 TI - On the origin of surface proteinase 3 of nonmyeloid cells: evidence favoring an exogenous source. AB - In Wegener's granulomatosis (WG), when the endogenous Proteinase 3 (PR3) of myeloid cells is translocated to the cell surface, a pathologically consequent interaction is believed to occur with classic anti-neutrophil cytoplasmic antibody (cANCA). In contrast, the exact origin of surface PR3 on cells of nonmyeloid origin is still debated. By various methods, PR3 mRNA and protein are easily demonstrated in myeloid cells but not in nonmyeloid cells. Exceptionally, the endothelial ECV304 cell line spontaneously produced PR3 mRNA but no PR3 protein. In the other nonmyeloid cells, we could not show cell surface PR3 either spontaneously or after TNFalpha stimulation. On the other hand, under serum-free conditions and using [(3)H]DFP-labeled HL-60 extract, a rapid, dose-dependent, saturable binding was demonstrated to both myeloid and nonmyeloid cells. That was reproduced with purified [(3)H]DFP-PR3. While we could not demonstrate cell surface PR3 on nonmyeloid cells after incubation with serum-containing supernatants of HL-60 cell cultures, we could do so after an overnight coculture period with HL-60 cell suspensions under the usual serum-containing culture conditions. Overall, our data would suggest that in vivo, the surface PR3 found on nonmyeloid cells is not endogenous but results from adsorption of PR3 extruded in their microenvironment by neighboring myeloid cells coming in close contact with them. PMID- 11027460 TI - Notice of duplicate publication. PMID- 11027459 TI - Immunodulation of rat serum and mucosal antibody responses to Entamoeba histolytica trophozoites by beta-1,3-glucan and cholera toxin. AB - Systemic and mucosal and immune responses can be manipulated with immunomodulators. Here we show the modulatory effects of cholera toxin (CT) and beta-1,3-glucan (GLU) on the rat antiamebic serum and fecal antibody responses to one or four intraperitoneal (IP) or intragastric (IG) doses of glutaraldehyde fixed Entamoeba histolytica trophozoites (GFT). One IP dose of GFT maximized serum IgM and IgG antiamebic antibodies on days 4 and 9, respectively; CT coadministration increased IgM antibodies, whereas IgG titers increased with CT or GLU; coproantibodies were undetectable after GFT alone or coadministered with GLU, whereas CT coadministration maximized fecal IgA antibodies on day 6. One IG dose of GFT alone increased serum IgM and IgG antibodies 2.5 times and no further increases were detected using GLU, whereas CT doubled serum IgG antibodies; GFT did not affect the coproantibody responses, whereas GLU coadministration maximized IgG coproantibody levels on day 6 and CT increased IgG and IgA coproantibody levels on the same day. On the other hand, four IG doses of GFT alone or with GLU induced tolerance, whereas GFT alone via the IP route increased serum antibodies slightly and GLU coadministration increased serum IgG antibody titers 300-fold. CT coadministration by both routes increased IgA coproantibodies, and simultaneous CT+GLU coadministration induced lower responses than either CT or GLU. Different antiamebic immune responses might therefore be attained through the use of different immunization routes and immunomodulators to induce protective immunity against intestinal or extraintestinal amebiasis. PMID- 11027461 TI - Prevention of suture hole bleeding using fibrin sealant: benefits of factor XIII. AB - BACKGROUND: Suture hole bleeding is a frequent complication in vascular surgery. The use of fibrin sealants (FSs) during surgery is reported to reduce blood loss by enhancing hemostasis. OBJECTIVES: Using a pig vascular graft model we investigated: (1) the use of FSs in vascular surgery and measured blood loss with and without the use of FSs; (2) the effect of factor XIII in FSs on hemostasis and blood loss; and (3) the effect of increasing FS incubation time on hemostasis and blood loss. MATERIALS AND METHODS: During surgery, a 3-cm incision was made in the carotid artery wall. Incisions were covered with a Gore-Tex (PTFE) patch sutured with Ethicon monofil 5/0 or 3/0. The resulting suture holes were treated according to study protocol. Study 1: Nine pigs received either FS treatment (n = 5) or no treatment (n = 4). Study 2: Twenty pigs underwent bilateral surgery; right (n = 20) and left (n = 20) patches received either FS with factor XIII (FS + FXIII) or FS depleted of factor XIII (FS - FXIII). Study 3: Bilateral surgery was carried out on 24 pigs; PFTE patches secured with Ethicon 3/0 were treated with FS and allowed to polymerize for 120 s (n = 12), 180 s (n = 12), or 240 s (n = 12) or with solid support/thrombin (n = 12). RESULTS: Study 1: Mean blood loss was significantly lower in the FS-treated group compared with untreated controls (9.2 +/- 10.6 mL vs 178.8 +/- 125.5 mL, mean +/- SD, P = 0.016). Study 2: Significantly less blood was lost in the FS + FXIII group than in the FS - FXIII group (Delta = 9.6 mL, P = 0. 02). Study 3: Blood loss was significantly higher from patches treated with solid support/thrombin compared with those treated with FS (P < 0.01). CONCLUSIONS: FSs containing factor XIII, such as Beriplast P, are effective in reducing suture hole bleeding and improving hemostasis during vascular graft procedures. The presence of factor XIII contributes to the efficacy of FSs and reduces the time to hemostasis. During vascular surgery, blood loss can be reduced significantly by the use of fibrin sealant compared with absorbable gelatin sponges coated with thrombin. PMID- 11027462 TI - Comparison of telomerase levels before and after differentiation of two cell lines of human neuroblastoma. AB - BACKGROUND: Telomerase is the enzyme that is responsible for maintaining telomere length in human germ cells, tumor cells, and immortalized cells. Its specific role in the immortilization process is unknown. This study was performed to determined whether the level of telomerase activity in human neuroblastoma cell lines correlated with their level of differentiation. We proposed that as neuroblastoma cells differentiated into more mature or benign cells, the levels of telomerase expression would decrease. MATERIALS AND METHODS: Two human neuroblastoma cells lines, SK-N-AS and SK-N-DZ, were differentiated using retinoic acid. These cells were assayed for telomerase activity by the telomere repeat amplification protocol (TRAP) before, during, and after treatment with retinoic acid for 8 days. Untreated cells were used for control and were compared to the retinoic acid-treated cells. Differentiation of the cell lines was confirmed by assaying expression of ret mRNA using the reverse-transcriptase polymerase chain reaction (RT-PCR) and gel electrophoresis of the radiolabeled products. RESULTS: No statistical difference in telomerase activity was noted between control and treated groups. CONCLUSIONS: While telomerase activity has been shown by others to correlate with tumor aggressiveness in human neuroblastoma cells, the mechanism that is involved appears to be separate from cellular differentiation. PMID- 11027463 TI - Significance of coexpression of urokinase-type plasminogen activator, and matrix metalloproteinase 3 (stromelysin) and 9 (gelatinase B) in colorectal carcinoma. AB - Our previous study showed that proMMP-9 was activated by MMP-3 directly, and that proMMP-3 was activated by plasmin. It was postulated that the proMMP-9 activation mechanism through the protease-protease cascade existed even in vivo. The purpose of the present study was to clarify the clinical significance of the combined expression of MMP-9, MMP-3, and urokinase-type plasminogen activator (uPA) in colorectal cancer, and the role of MMP-3 or uPA expression as an activator for MMP-9. The expression of both MMP-9 and uPA was found to be correlated with liver metastasis, and with survival rate. The coexpression of MMP-9 and uPA by tumor cells was also significantly correlated with postoperative hepatic recurrence and survival rate. MMP-9 tended to be coexpressed with uPA, and was consistently associated with MMP-3 localized at the tumor-invasive front with inflammatory cells such as monocyte-macrophages. In gelatin zymography, the MMP-9 active form tended to be identified in the tumors that coexpressed both MMP-9 and uPA. We concluded that coexpression of MMP-9 and uPA in tumor tissues might be a useful predictive factor for postoperative survival and hepatic metastasis. The following activation mechanism for proteinase might occur: uPA coexpressed with MMP-9 activated plasminogen, and plasmin activated proMMP-3, which was secreted depending upon inflammatory infiltration, and then MMP-3 activated proMMP-9, resulting in colorectal cancer progression and metastasis. PMID- 11027464 TI - Angiogenic therapy for the chronically ischemic lower limb in a rabbit model. AB - The purpose of this study was to evaluate the long-term effectiveness of basic fibroblast growth factor (bFGF) in achieving neovascularization following ischemia from arterial ligation and to determine an optimal dosage level. We used an Ameroid constrictor to produce progressive occlusion of the left femoral artery of rabbits. At 2 weeks, the rabbits were randomized to receive intravenous injection of vehicle (group A, n = 15); 3 microg/kg/day bFGF (group B, n = 12); 10 microg/kg/day bFGF (group C, n = 12); or 16 microg/kg/day bFGF (group D, n = 15) for 3 days. At 1 to 37 days after surgery, we assessed limb neovascularization by transcutaneous oximetry (TCPO(2)), angiography, heart rate, arterial pressure, peripheral vascular resistance (PRU), and muscle blood flow (MBF) during steady-state intra-arterial infusion of saline (basal), acetylcholine, papaverine, or serotonin under anesthesia and capillary density (cap/mm(2)) and capillary per muscle fiber ratio (cap/F). Groups B and C showed significantly greater change in TCPO(2) over time than groups A and D (P < 0.0001). Group D showed the lowest TCPO(2) values from days 14 to 37 and group C the highest. Groups B and C showed a higher number of vessels filled with contrast agent than groups A and D (P < 0.0001). Calf cap/mm(2) and cap/F were significantly higher in groups B and C than groups A and D (P < 0.0001). Calf basal MBF values were higher in groups B and C than in groups A and D, but were not statistically significant. Group D showed the highest level in basal PRU. There were no significant differences in heart rate or blood pressure among the groups. These results show (1) treatment with bFGF has no adverse hemodynamic effects, (2) bFGF enhances angiogenesis and circulation at moderate doses, and these effects persist at least several weeks, and (3) high doses of bFGF may inhibit angiogenesis and collateral circulation. PMID- 11027465 TI - Collagen-embedded platelet-derived growth factor DNA plasmid promotes wound healing in a dermal ulcer model. AB - BACKGROUND: Gene therapy has shown limited efficacy for treating congenital diseases, partly due to temporary gene expression and host immune responses. Such results suggest that gene therapy is ideal for chronic wound treatment where limited duration of target gene expression is required. This study tested the wound healing effects of topically applied platelet-derived growth factor (PDGF) A or -B chain DNA plasmids embedded within a collagen lattice. MATERIALS AND METHODS: Four 6-mm dermal ulcer wounds were created in the ears of young adult New Zealand White rabbits made ischemic by division of the central and rostral arteries. Wounds were treated with lyophilized collagen containing PDGF-B DNA (1.0-3.0 mg), PDGF-A DNA (1.0 mg), irrelevant DNA (1.0 mg), or collagen alone. Wounds were dressed and harvested after 10 days for measurement of granulation tissue formation, epithelialization, and wound closure. Results were evaluated with a paired two-tailed Student t test, with P values < 0.05 considered significant. RESULTS: PDGF-B DNA increased new granulation tissue (NGT) formation up to 52% and epithelialization 34% compared with controls. Wound closure was increased up to threefold. At 1.0 mg DNA, PDGF-A and PDGF-B stimulated similar responses. No difference in NGT or epithelialization was seen between control groups. CONCLUSIONS: PDGF DNA gene therapy is effective at accelerating wound healing in ischemic dermal ulcers and provides a viable alternative to peptide growth factor therapy. PMID- 11027466 TI - Optimization of rat hepatocyte culture in citrated human plasma. AB - BACKGROUND: Maintenance of liver-specific functions in hepatocyte cultures during plasma exposure is critical for the clinical application of bioartificial liver assist systems. Sodium citrate is a common anticoagulant but has been shown to be cytotoxic to hepatocytes. We have tested the effect of various supplements on the viability and function of adult primary rat hepatocytes exposed to citrated plasma. MATERIALS AND METHODS: Freshly isolated rat hepatocytes were cultured in the collagen gel sandwich configuration in culture medium for 6 days followed by exposure to citrated human plasma with various supplements for 1 week. Controls were left in culture medium throughout. Viability and synthetic functions were evaluated. RESULTS: Hepatocytes exposed to unsupplemented citrated plasma lost significant viability and function within the first 2 days. Cells cultured in plasma supplemented with a fivefold concentrate of standard hepatocyte culture medium maintained urea (1. 2-2.1 micromol/day/10(6) cells) and albumin (51-62 microg/day/10(6) cells) synthesis rates equal to or higher than those of controls. Among the various components of the concentrated medium supplement, calcium chloride (1.8 mM), magnesium sulfate (0.8 mM), amino acids (fourfold Basal Medium Eagle amino acids including 4 mM glutamine), and glucagon (14 ng/ml) were found to be essential in maintaining urea synthesis. Maintenance of a high albumin synthesis rate also required the addition of hydrocortisone (7.5 microg/ml) and insulin (0.5 U/ml). CONCLUSIONS: Appropriate metabolic and hormonal supplementation of citrated human plasma prevents its cytotoxic effects and may be used in conjunction with in vivo use of bioartificial liver assist systems. PMID- 11027467 TI - Role of nitric oxide in hemorrhagic shock-induced bacterial translocation. AB - BACKGROUND: Hemorrhagic shock-induced bacterial translocation is an etiologic factor in the pathogenesis of multiple system organ damage. Excessive production of nitric oxide (NO) during hemorrhagic shock may lead to cellular injury and gut barrier failure that promotes bacterial translocation. We investigated the effect of aminoguanidine (AG) and N(G)-nitro-l-arginine methyl ester (l-NAME), both inhibitors of NO synthase, on hemorrhagic shock- induced bacterial translocation in the rat. MATERIALS AND METHODS: Anesthetized male Sprague-Dawley rats were subjected to a hemorrhagic shock protocol for 30 min followed by intravenous injection (1 mL/kg body wt) with normal saline, AG (100 mg/kg), or l-NAME (10 mg/kg). Tissues/organs were examined histologically for damage and bacterial translocation. Plasma nitrate/nitrite was measured using a procedure based on the Griess reaction, and nitric oxide synthase (NOS) expression was determined immunohistochemically. RESULTS: The shocked animals treated with saline died within 90 min, and deaths were associated with 100% bacterial translocation, increased tissue/organ damage, and elevated nitrate/nitrite production. In contrast, both AG and l-NAME increased the survival time of shocked rats to >72 h, abrogated bacterial translocation, reduced tissue/organ damage, and prevented excessive nitrate/nitrite production and upregulation of expression of endothelial NOS and inducible NOS. CONCLUSIONS: Prevention of bacterial translocation by pharmacologic agents such as aminoguanidine and l-NAME could be an important therapeutic approach to lessen mortality rates following hemorrhagic shock. PMID- 11027468 TI - Effects of tumor necrosis factor-binding protein on hepatic protein synthesis during chronic sepsis. AB - BACKGROUND: Cytokines are thought to play a role in the stimulation of protein synthesis in liver during inflammation and sepsis. We previously showed that administration of tumor necrosis factor-binding protein (TNFbp) prevents the sepsis-induced inhibition of protein synthesis in skeletal muscle. The purpose of the present set of experiments was to investigate the effect of TNFbp on hepatic protein synthesis and its ability to modulate the mechanisms responsible for increased hepatic protein synthesis during chronic (5-day) intraabdominal sepsis. MATERIALS AND METHODS: We examined the effects of TNFbp on hepatic protein synthesis during sepsis in four groups of rats: control, control + TNFbp, septic, and septic + TNFbp. Saline (1.0 ml) or TNFbp (1 mg/kg, 1.0 ml) was injected daily starting 4 h prior to the induction of sepsis. The effect of sepsis and TNFbp administration on hepatic protein synthesis in vivo was examined 5 days later. RESULTS: Sepsis increased the rate of protein synthesis by 35% relative to controls. Accelerated rates of protein synthesis were accompanied by increased total RNA content, eukaryotic initiation factor (eIF) 2alpha content, and phosphorylation of p70S6 kinase. Injection of TNFbp into septic rats for 5 days did not diminish the sepsis-induced stimulation of hepatic protein synthesis. Compared with controls, septic rats treated with TNFbp also showed elevated total RNA content, elF2alpha content, and phosphorylation of p70S6 kinase. No significant differences in any of the parameters measured were observed between untreated and TNFbp-treated septic rats. Treatment of control animals with TNFbp for 5 days was without effect on any of the parameters examined. CONCLUSIONS: TNFbp did not prevent the sepsis-induced stimulation of hepatic protein metabolism or modulate the septic-induced changes in factors regulating protein synthesis. Global rates of protein synthesis in livers from septic rats are accelerated by increases in the abundance or activity of components of translational apparatus. PMID- 11027470 TI - Effects of exogenous endothelin-1 application on liver perfusion in native and transplanted porcine livers. AB - PURPOSE: This study was designed to assess and differentiate the impact of progressivly increasing portal venous endothelin-1 (ET) plasma concentrations on hepatic micro- and macroperfusion of native porcine livers (Group A) and liver grafts after experimental transplantation (Group B). METHODS: A standardized gradual increment in systemic ET plasma concentration (0-58 pg/ml) was induced by continuous ET-1 infusion into the portal vein in both groups (A: n = 10, B: n = 10). Control animals received only saline (n = 5, each group). Hepatic microcirculation (HMC) was quantified by thermodiffusion electrodes, hepatic artery flow (HAF), and portal venous flow (PVF) by Doppler flowmetry. RESULTS: No changes in ET or perfusion parameters were observed in controls. The mean ET level after orthotopic liver transplantation (OLT) in Group B was elevated (baseline: 3.8 +/- 2.4 pg/ml) compared with Group A (2.8 +/- 1.9 pg/ml). With rising ET levels HAF decreased progressively in Group A from 205 +/- 97 (baseline) to 160 +/- 72 ml/min, and in Group B from 161 +/- 87 to 146 +/- 68 ml/min. PVF decreased in Group A from 722 +/- 253 to 370 +/- 198 ml/min, and in Group B from 846 +/- 263 to 417 +/- 203 ml/min. Baseline HMC in Group A was 86 +/ 15 and decreased significantly to 29 +/- 9 ml/100 g/min, and baseline MC in Group B was 90 +/- 22 and decreased to 44 +/- 32 ml/100 g/min. No significant alteration in systemic circulation was noted at the ET concentrations investigated. CONCLUSIONS: Significant impairment of hepatic micro- and macrocirculation was detected after induction of systemic ET levels above 9.4 pg/ml both in native and in transplanted livers. Disturbance of HMC was caused predominantly by reduction of portal venous flow, while the effect of ET on HAF was less pronounced. Characteristics of flow impairment in transplanted and native livers were analogous after short cold ischemic graft storage (6 h). PMID- 11027469 TI - Role of platelet-activating factor in leukocyte-independent plasma extravasation and mast cell activation during endotoxemia. AB - BACKGROUND: Independently from leukocyte adherence, endothelial factors and mast cell activation seems to promote microvascular permeability. Platelet-activating factor (PAF) has been shown to play a significant role in endotoxin-induced leukocyte adherence. The aim of our study was to investigate if there is also a role for PAF in mediating leukocyte-independent microvascular permeability changes and activation of mast cells during endotoxemia. Therefore, during endotoxemia microvascular permeability and mast cell activation were determined after inhibition of L-selectin-mediated leukocyte adherence by fucoidin and after inhibition of PAF effects by the PAF receptor antagonist BN52021. MATERIALS AND METHODS: In male Wistar rats, red cell velocity (V(RBC)), venular wall shear rate, microvascular permeability, leukocyte adherence, and mast cell activation were determined in mesenteric postcapillary venules using intravital microscopy at baseline and 60 and 120 min after start of a continuous infusion of endotoxin (ETX; 2 mg/kg/h, Escherichia coli O26:B6) (ETX group). Animals in the FUCO/ETX group received fucoidin (25 mg/kg body wt) in addition to the procedure described above. Animals in the FUCO/ETX/PAF-ANT group received fucoidin and the PAF receptor antagonist BN52021 (5 mg/kg body wt) prior to the continuous endotoxin infusion. Control animals (control group) received only equivalent volumes of NaCl 0.9%. RESULTS: There were no microhemodynamic and macrohemodynamic differences between groups. In all endotoxin-challenged groups macromolecular leakage and mast cell activity increased significantly, starting at 60 min. Both macromolecular leakage and mast cell activity were significantly higher in the FUCO/ETX group than in the FUCO/ETX/PAF-ANT group and control group. Differences in macromolecular leakage between groups were significant at 120 min. Differences in mast cell activity between groups were significant at 60 and 120 min. CONCLUSIONS: The results of our study demonstrate a leukocyte-independent plasma extravasation that can be inhibited by the PAF receptor antagonist BN52021, indicating the involvement of PAF in the pathophysiology of leukocyte-independent microvascular damage during early endotoxemia. Mast cell activity seems to precede leukocyte-independent macromolecular leakage. PMID- 11027471 TI - The association for academic surgery PMID- 11027472 TI - Association for academic surgery 34th annual meeting PMID- 11027474 TI - AAS the association for academic surgery newsletter PMID- 11027473 TI - AAS 2000 membership list PMID- 11027476 TI - On selection mechanisms during initial evolution and on the possible role of ionic balance. AB - Initial evolution at the molecular level can proceed through mechanisms that enhance either the generation or the survival of certain molecules. Phenomena related to ionic balance can be the basis for both generative and survival selection mechanisms that may have driven the initial evolution of macromolecules. An important advantage of mechanisms based on ionic balance phenomena is that they can conceivably generate spatial and temporal inhomogeneities in semi-enclosed spaces within cavities of porous materials and/or membranes. Under certain circumstances, such inhomogeneities are required for initial evolution to proceed. PMID- 11027475 TI - Mathematical treatment of transport data of bacterial transport systems to estimate limitation in diffusion through the outer membrane. AB - Bacterial transport systems are traditionally treated as enzymes exhibiting a saturable binding site giving rise to an apparent K(m)of transport, whereas the maximal rate of transport is regarded equivalent to the V(max)of enzymatic reactions. Thus, the Michaelis-Menten theory is usually applied in the analysis of transport data and K(m)and V(max)are derived from the treatment of data obtained from the rate of transport at varying substrate concentrations. Such an analysis tacitly assumes that the substrate recognition site of the transport system is freely accessible to substrate. However, this is not always the case. In systems endowed with high affinity in the micro M range or those recognizing large substrates or those exhibiting high V(max), the diffusion through the outer membrane may become rate determining, particularly at low external substrate concentrations. In such a situation the dependence of the overall rate of transport (from the medium into the cytoplasm) on the substrate concentration in the medium will no longer follow Michaelis-Menten kinetics. By analysing the deviation of transport data from the corresponding ideal Michaelis-Menten plot we developed a method that allows us to determine diffusion limitation through the outer membrane. The method allows us to find the correct K(m)of the transport system functioning at the inner membrane even under conditions of strong diffusion limitation through the outer membrane. The model was tested and validified with the Escherichia coli binding protein-dependent ABC transporter for maltose. The corresponding systems for sn -glycerol-3-phospate of Escherichia coli and the alpha -cyclodextrin transport of Klebsiella oxitoca were used as test systems. PMID- 11027477 TI - Learning rules for social foragers: implications for the producer-scrounger game and ideal free distribution theory. AB - In population games, the optimal behaviour of a forager depends partly on courses of action selected by other individuals in the population. How individuals learn to allocate effort in foraging games involving frequency-dependent payoffs has been little examined. The performance of three different learning rules was investigated in several types of habitats in each of two population games. Learning rules allow individuals to weigh information about the past and the present and to choose among alternative patterns of behaviour. In the producer scrounger game, foragers use producer to locate food patches and scrounger to exploit the food discoveries of others. In the ideal free distribution game, foragers that experience feeding interference from companions distribute themselves among heterogeneous food patches. In simulations of each population game, the use of different learning rules induced large variation in foraging behaviour, thus providing a tool to assess the relevance of each learning rule in experimental systems. Rare mutants using alternative learning rules often successfully invaded populations of foragers using other rules indicating that some learning rules are not stable when pitted against each other. Learning rules often closely approximated optimal behaviour in each population game suggesting that stimulus-response learning of contingencies created by foraging companions could be sufficient to perform at near-optimal level in two population games. PMID- 11027478 TI - Predicting response to selection on a quantitative trait: a comparison between models for mixed-mating populations. AB - Two different theoretical frameworks have been developed to predict response to selection in a mixed mating population (in which reproduction occurs by a mixture of outcrossing and self-fertilization). The genotypic covariance model (GCM) and the structured linear model (SLM) rely on the same assumptions regarding quantitative trait inheritance, but use different genetic summary statistics. Here, we demonstrate the algebraic relationships between the various genetic metrics used in each theory. This is accomplished by reformulating the GCM in terms of the Wright-Kempthorne equation. We use stochastic simulations to investigate the relative accuracy of each theory for a range of selfing rates. The SLM is generally more accurate than the GCM, the most pronounced differences emerging in simulations with inbreeding depression for fitness. In fact, with strong inbreeding depression and high selfing rates, evolution can occur opposite the direction predicted by the GCM. The simulations also indicate that direct application of random mating models to partially selfing populations can produce very inaccurate predictions if quantitative trait loci exhibit dominance. PMID- 11027479 TI - Using Shannon entropy on measuring the individual variability in the Rufous bellied thrush Turdus rufiventris vocal communication. AB - We applied the information theory concepts to notes repertoire characteristics combined with temporal parameters of the Rufous-bellied thrush Turdus rufiventris song, using this particular case to test a new method of analysing quantitatively complex animal communication systems. Like most Turdus thrushes, Rufous-bellied thrushes are remarkable for their long, varied and melodious songs. For the analysis of the species repertoire, we used recordings of 44 individuals from 24 localities covering its full geographical range. We measured the repertoire size, note duration and rhythm (frequency of note utterance), and combined these parameters with the Shannon entropy values calculated for each individual. Although individuals maintain species-specific recognition capacity, we find a large variation between their song parameters and show that the information theory can be useful to analyse large and varied animal vocal repertoires. We are introducing two new parameters, temporal average entropy (E(t)) and utterance frequency average entropy (E(f)), for measuring such communication systems. PMID- 11027480 TI - The role of cytotoxic T lymphocytes in human T-cell lymphotropic virus type 1 infection. AB - The vast majority of individuals infected with human T-cell lymphotropic virus type 1 have a strong and persistently activated cytotoxic T lymphocyte response to the virus. Experimental work investigating the effects of human T-cell lymphotropic virus-specific cytotoxic T lymphocytes is conflicting. One significant body of work suggests that specific cytotoxic T lymphocytes are protective and help to reduce the risk of disease. However, another body of work implies that specific cytotoxic T lymphocytes play an important role in the development of disease. Here we use a theoretical model to explore the role of cytotoxic T lymphocytes in persistent infection. A way of reconciling the apparently contradictory data is proposed and experimentally testable predictions are made. PMID- 11027481 TI - Species-area curves, spatial aggregation, and habitat specialization in tropical forests. AB - The relationship between species diversity and sampled area is fundamental to ecology. Traditionally, theories of the species-area relationship have been dominated by random-placement models. Such models were used to formulate the canonical theory of species-area curves and species abundances. In this paper, however, armed with a detailed data set from a moist tropical forest, we investigate the validity of random placement and suggest improved models based upon spatial aggregation. By accounting for intraspecific, small-scale aggregation, we develop a cluster model which reproduces empirical species-area curves with high fidelity. We find that inter-specific aggregation patterns, on the other hand, do not affect the species-area curves significantly. We demonstrate that the tendency for a tree species to aggregate, as well as its average clump size, is not significantly correlated with the species' abundance. In addition, we investigate hierarchical clumping and the extent to which aggregation is driven by topography. We conclude that small-scale phenomena such as dispersal and gap recruitment determine individual tree placement more than adaptation to larger-scale topography. PMID- 11027482 TI - A plausible mechanism of electron transfer between quinones in photosynthetic reaction centers. AB - The mechanism of long-range electron transfer between the primary and the secondary quinone of photosynthetic reaction centers has been investigated, with particular attention on the role of the iron-histidine bridge. Computations suggest that in such a system, where the molecular subunits are packed together by H-bonds, a mobile electron, injected on one end of the chain, can be carried to the other end by switching the positions of the H-bonded hydrogens. Energy estimates would suggest that the proposed mechanism is plausible and worthy of further experimental investigations. PMID- 11027483 TI - A model of pattern formation based on signaling pathways. AB - A model of pattern formation in the early embryo is presented. It is motivated by the necessary interaction of kinases and phosphatases, and in particular by the family of Wnt kinase and RPTP phosphatase signaling pathways. It is seen as complementary to the more short-range patterning of the Delta/Notch (or juxtacrine) signaling pathway. PMID- 11027484 TI - Do evolutionary processes minimize expected losses? AB - Evolution by variation and natural selection is often viewed as an optimization process that favors those organisms which are best adapted to their environment. This leaves open the issue of how to measure adaptation and what criterion is implied for optimization. This problem has been framed and analysed mathematically under the assumption that individuals compete to minimize expected losses across a series of decisions (e.g. choice of behavior), where each decision offers a stochastic payoff. But the fact that a particular analysis is tractable for a specified criterion does not imply the fidelity of that criterion. Computer simulations involving a version of the k -armed bandit problem can address the veracity of the hypothesis that individuals are selected to minimize expected losses. The results offered here do not support this hypothesis. PMID- 11027485 TI - Multiple steady states in kinetic models of red cell metabolism. PMID- 11027486 TI - Is the unfolded state the Rosetta Stone of the protein folding problem? AB - Solving the protein folding problem is one of the most challenging tasks in the post genomic era. Identification of folding-initiation sites is very important in order to understand the protein folding mechanism. Detection of residual structure in unfolded proteins can yield important clues to the initiation sites in protein folding. A substantial number of studied proteins possess residual structure in hydrophobic regions clustered together in the protein core. These stable structures can work as seeds in the folding process. In addition, local preferences for secondary structure in the form of turns for beta-sheet initiation and helical turns for alpha-helix formation can guide the folding reaction. In this respect the unfolded states, studied at increasing structural resolution, can be the Rosetta Stone of the protein folding problem. PMID- 11027487 TI - Fibroblast growth factor (FGF)-2 mediates cell attachment through interactions with two FGF receptor-1 isoforms and extracellular matrix or cell-associated heparan sulfate proteoglycans. AB - In the presence of FGF-2, cells in suspension expressing FGF receptor-1 will attach to monolayers of cells expressing heparan sulfates. This attachment provides physical evidence for the formation of a trimolecular complex between FGF-2, heparan sulfate, and FGF receptors. We have used this system to determine if receptor isoforms containing or lacking the first of three immunoglobulin-like domains are equally able to form complexes with FGF-2 and heparan sulfates. In the presence of FGF-2, cells expressing either isoform of the receptor were able to attach to monolayers of CHO cells expressing heparan sulfates. No attachment was observed in the absence of FGF-2 or if heparin was included in the incubation medium. Attachment of cells expressing the two receptor isoforms occurred at similar concentrations of FGF-2, and similar concentrations of heparin were required to disrupt the interactions. Thus, there appeared to be little difference between these receptor isoforms in their ability to form trimolecular complexes with FGF-2 and cell-associated heparan sulfates. We also found that, in the presence of FGF-2, cells expressing FGF receptor-1 are able to form complexes with both extracellular matrix and cell-surface heparan sulfates. PMID- 11027488 TI - Differential regulation of COX-2 transcription by Ras- and Rho-family of GTPases. AB - Cyclooxygenase-2 (Cox-2) gene expression which is rapidly induced by cytokines, growth factors and tumor promoters, is important for inflammation, angiogenesis, and is markedly enhanced in various cancer cells. Many of these factors initiate signaling through Ras- and Rho-family small GTPases. Here, we investigated the ability of Ras, Rac, Rho, and Cdc42Hs to differentially regulate transcription from the murine COX-2 promoter in NIH 3T3 cells. Over-expression of constitutively active mutants of Ras, Rac, Rho, but not Cdc42Hs induced transcription from the COX-2 promoter. Transactivation by Rac and Rho required cis-acting elements located between -80 and -40 of the COX-2 promoter whereas deletion of this region enhanced transactivation by Ras. A CRE/ATF element located at -56 was critical for Ras- and Rac-induced transactivation of the COX-2 promoter, but was not required for transactivation by Rho. This demonstrates Rho dependent transactivation of the COX-2 promoter through novel trans-acting elements and suggests that, in NIH 3T3 cells, signaling by small GTPases that result in COX-2 expression is not through a sequential pathway from Cdc42 to Rac to Rho, but rather through independent, parallel signaling pathways. PMID- 11027489 TI - Cloning and characterization of the 5'-flanking region of the human prolactin releasing peptide receptor gene. AB - Recently a novel peptide which specifically stimulates the secretion of prolactin (PRL) was found and named PRL-releasing peptide (PrRP). To evaluate the regulation of human (h) PrRP-receptor (PrRP-R) gene expression, we cloned the 5' flanking region of the hPrRP-R gene and determined the nucleotide sequence of 4.0 kilobase pairs (kb) upstream from the translation start site. Analysis of the hPrRP-R transcripts by means of 5'-rapid amplification of cDNA ends suggested that the hPrRP-R gene had multiple transcription start sites between -429 and 365 from the translation start site. There is no typical TATA or CAAT but a GC box and putative binding sites for several transcription factors including Pit-1 and pituitary homeobox 1 (Ptx1). However, transient transfection studies using a luciferase reporter gene demonstrated that 5'-flanking region exerts promoter activity in several non-pituitary cell lines as well as in GH(3) cells. The GC box located from -467 to -457 was identified as an important region for the basal expression of the hPrRP-R gene. Knowledge of the promoter region of the hPrRP-R gene, which was obtained in the present study, will facilitate the clarification of its transcriptional regulation. PMID- 11027490 TI - Rescue of CH31 B cells from antigen receptor-induced apoptosis by inhibition of p38 MAPK. AB - CH31 B lymphomas represent a model for antigen-induced deletional tolerance of immature B lymphocytes, because cross-linking the B cell antigen receptor (BCR) induces G(1) phase arrest and apoptosis. We have recently demonstrated that BCR cross-linking leads to a transient activation of p38 mitogen-activated protein kinase (MAPK) in CH31 B cells. In this paper, we functionally characterize the role of p38 MAPK in BCR-induced apoptosis as well as evaluate the regulation of additional MAPKs by the BCR. We demonstrate that JNK and ERK activities are not affected by BCR cross-linking, suggesting that these MAPKs are not directly involved in initiating the apoptotic cascade. By contrast, we show that pretreatment of CH31 B cells with the highly specific p38 MAPK inhibitor SB203580 ablated both BCR-induced p38 MAPK activity and apoptosis. Pretreatment of CH31 cells with an inactive SB203580 analog, SB202474, did not prevent apoptosis. These findings establish a key role for p38 MAPK in antigen receptor-mediated apoptosis of CH31 B cells. PMID- 11027491 TI - Amyloid beta protein starting pyroglutamate at position 3 is a major component of the amyloid deposits in the Alzheimer's disease brain. AB - The amyloid beta protein (Abeta) deposited in the Alzheimer's disease (AD) brain is heterogeneous at both its amino and carboxyl termini. Recent studies of the genetic forms of AD indicate that the aggregation and deposition of Abeta42 may be a common initiating event in all forms of AD. Here, we analyzed the amino termini of the Abeta species deposited in the AD brain, focusing specifically on species with amino-terminal pyroglutamate at position 3 (Abeta3(pE)). Immunocytochemical analysis of AD brains with an antibody specific for Abeta3(pE) confirmed that these species deposit in blood vessels and senile plaques. Using specific sandwich ELISAs, we determined the amounts of Abeta3(pE)-40 and Abeta3(pE)-42(43) in AD brain compared with other forms. This analysis showed that Abeta3(pE)-40 is closely correlated with the extent of Abeta deposition in blood vessels, whereas Abeta3(pE)-42(43) is not. In addition, Abeta3(pE)-42(43) is an important component of the Abeta deposited in senile plaques of the AD brain, constituting approximately 25% of the total Abeta42(43). In vitro comparison of Abeta1-42 and Abeta3(pE)-42 showed that Abeta3(pE)-42 is highly prone to oligomerization. These findings suggest that Abeta3(pE)-42 may be particularly important in AD pathogenesis. PMID- 11027492 TI - Human 11beta-hydroxysteroid dehydrogenase type 1 is enzymatically active in its nonglycosylated form. AB - 11beta-Hydroxysteroid dehydrogenase type 1 (11beta-HSD 1) is a microsomal enzyme responsible for the reversible interconversion of active 11beta hydroxyglucocorticoids into inactive 11-ketosteroids and by this mechanism regulates access of glucocorticoids to the glucocorticoid receptor. The enzyme has also been proven to participate in xenobiotic carbonyl compound detoxification. 11beta-HSD 1 is anchored within the membranes of the endoplasmic reticulum (ER) by its N-terminus, whereby its active site protrudes into the lumen of the ER. In the primary structure of 11beta-HSD 1 three Asn-X-Ser glycosylation motifs have been identified. However, the importance of N-linked glycosylation of 11beta-HSD 1 for catalytic activity has been controversely discussed. To clarify if glycosylation is essential for enzyme activity, we performed deglycosylation experiments of native 11beta-HSD 1 from human liver as well as site-directed mutagenesis to remove potential glycosylation sites upon overexpression in Pichia pastoris. The altered proteins were examined regarding their catalytic activity towards their physiological glucocorticoid substrates. The molecular size of the various 11beta-HSD 1 forms was analyzed by immunoblotting with a polyclonal antibody raised against 11beta-HSD 1 protein from human liver. By stepwise enzymatic deglycosylation of native 11beta-HSD 1 we could demonstrate that all potential glycosylation sites carry N-linked oligosaccharide residues under physiological conditions. Interestingly, complete deglycosylation did not affect enzyme activity, neither in the reductive (cortisone) nor in the oxidative (cortisol) direction. Upon overexpression in the yeast P. pastoris, 11beta-HSD 1 did not undergo glycosylation, but, in spite of this, yielded a fully active enzyme. Our results conclusively demonstrate that 11beta-HSD 1 does not need to be glycosylated to perform its physiological role as glucocorticoid oxidoreductase. PMID- 11027493 TI - Purification and identification of neuromedin U as an endogenous ligand for an orphan receptor GPR66 (FM3). AB - GPR66 is an orphan G-protein-coupled receptor (GPCR) whose structure is similar to the ghrelin and motilin receptors. We have tried to purify a natural ligand for GPR66 in rat tissues and identified a 23-amino-acid peptide as the endogenous ligand. Sequence analysis revealed the peptide as neuromedin U (NMU), a smooth muscle-contracting peptide that was first purified from porcine spinal cord by our group. NMU binds to GPR66-expressing cells with high specificity to induce intracellular calcium mobilization. When NMU was injected intracerebroventricularly (ICV) into rats, it potently suppressed food intake. In contrast, ICV injection of NMU-antibody increased food intake. These results suggest that NMU is a potent endogenous anorexic peptide. PMID- 11027494 TI - Identification of a novel human mitochondrial D-loop RNA species which exhibits upregulated expression following cellular immortalization. AB - We report the identification and characterization of a novel human mitochondrial RNA species approximately 0.47 kb long that is transcribed from the mtDNA L strand and is derived from the D-loop. Its expression increases when human cells become immortal, a key event in tumorigenesis. The RNA is therefore designated IDL (Immortalization-associated D-Loop). Sequence and hybrid cell analyses suggest that the increased level of IDL RNA in immortal cells is due to a recessive change, possibly in the activity of a trans-acting factor that controls IDL RNA expression. PMID- 11027495 TI - Inhibition of interleukin-6 signaling and Stat3 activation by a new class of bioactive cyclopentenone derivatives. AB - The IL-6-dependent activation of the JAK/STAT pathway plays a central role in the induction of the acute phase response in the liver. In a search for new inhibitors of the IL-6-mediated signal transduction in HepG2 cells using secreted alkaline phosphatase (SEAP) as reporter gene, four novel cyclopentenones, 2-(1 chloropropenyl)-4,5-dihydroxycyclopent-2-enone (CPDHC, 1), 4, 5-dihydroxy-2 propenylcyclopent-2-enone (DHPC, 2), 5-hydroxy-2, 3-dimethylcyclopent-2-enone (HDC, 3), and 4-methyl-5-methylenecyclopent-3-en-1,2-diol (MMCD, 4) were isolated from fermentations of the ascomycete strain A23-98. CPDHC (1) inhibits the IL-6 induced SEAP expression with IC(50) values of 4. 0-5.3 microM (0.75-1 microg/ml). The compounds DHPC (2), HDC (3), and MMCD (4) which are structurally closely related to CPDHC (1) showed no inhibitory effects on the IL-6-induced SEAP expression in HepG2 cells. Studies on the mode of action revealed that CPDHC (1) affects the IL-6-dependent pathway by inhibiting the tyrosine phosphorylation of the STAT3 and STAT1 as well as the serine phosphorylation of the Stat3 transcription factor. In addition, CPDHC (1) and DHPC (2) inhibit the AP-1 and NF kappaB mediated SEAP expression in transiently transfected HeLa S3 cells with IC(50) values of 10-15 microM (2-3 microg/ml) and 50-100 microM (8-16 microg/ml) respectively. Our results indicate that CPDHC inhibits the NF-kappaB pathway by preventing the phosphorylation and proteolytic degradation of the IkappaBalpha protein. The novel cyclopentenones may represent lead compounds for the development of new anti-inflammatory drugs. PMID- 11027496 TI - Genomic organization and amplification of the human desmosomal cadherin genes DSC1 and DSC3, encoding desmocollin types 1 and 3. AB - The desmosomal cadherins comprise the desmocollins and desmogleins and are involved in epithelial cell-cell adhesion. There are three desmocollins (DSC 1-3) and three desmogleins (DSG 1-3) that are expressed in a tissue- and development specific manner. Desmosomal proteins have been implicated in a number of disorders characterized by loss of cell-cell adhesion and trauma-induced skin fragility. Therefore, the desmocollins are potential candidates for genodermatoses involving epithelial tissues. In order to screen the entire DSC1 and DSC3 genes, we have characterized their intron-exon organization. The DSC1 gene comprises 17 exons spanning approximately 33 kb on 18q12.1, and the DSC3 gene comprises 17 exons spanning approximately 49 kb on 18q12.1. We have also developed a comprehensive PCR-based mutation detection strategy for desmocollins 1, 2, and 3 using primers placed on flanking introns followed by direct sequencing of the PCR products. PMID- 11027497 TI - Ionic products of bioactive glass dissolution increase proliferation of human osteoblasts and induce insulin-like growth factor II mRNA expression and protein synthesis. AB - Bioglass 45S5 is an osteoproductive material, which resorbs by releasing its constitutive ions into solution. Treatment with the ionic products of Bioglass 45S5 dissolution in DMEM for 4 days increased human osteoblast proliferation to 155% of control. Two days after treatment, differential gene expression was analyzed by cDNA microarrays. Expression of a potent osteoblast mitogenic growth factor, insulin-like growth factor II (IGF-II), was increased to 290%. Additionally, there was a 168% increase in the concentration of unbound IGF-II protein in the conditioned media of treated osteoblasts. Expression levels of IGFBP-3, an IGF-II carrier protein, metalloproteinase-2 and cathepsin-D were also increased to 200, 340, and 310% of control levels, respectively. Metalloproteinase-2 and cathepsin-D are proteases that cleave IGF-II from its carrier proteins, resulting in the release of the unbound biologically active IGF II. We suggest that the stimulatory effect of the ionic products of Bioglass 45S5 dissolution on osteoblast proliferation may be mediated by IGF-II. PMID- 11027498 TI - Isolation and molecular characterization of the 5'-upstream region of the human TRAIL gene. AB - TRAIL, a novel member of the TNF family, acts through membrane receptors to induce apoptosis of activated T lymphocytes and may represent a mechanism for the "immune escape" of certain cancers. Various cytokines appear to increase expression of other TNF family members; however, the regulation of TRAIL has not been defined. The purpose of this study was to assess molecular mechanisms regulating TRAIL gene expression in human colon cancers. In this study, we have cloned the human TRAIL (hTRAIL) promoter ( approximately 1.6 kb) and identified a number of putative transcription factor binding sites such as NFAT, AP-1 and Sp1 sequences which are important for the expression of other TNF family members. Transient transfections of 5'-deletion promoter constructs into either Caco-2 or HT29 colon cancer cells identified TRAIL promoter regions critical for both basal and interferon-gamma (IFN-gamma)-mediated induction. Furthermore, induction of TRAIL mRNA levels was demonstrated in HT29 and Caco-2 cells with IFN-gamma treatment suggesting an important role for this cytokine in TRAIL expression. PMID- 11027499 TI - Protein kinase C-alpha participates in FcgammaR-mediated phagocytosis in macrophages. AB - Phagocytosis of IgG-opsonized particles by macrophages requires the activation of protein kinase C (PKC), a family of protein serine/threonine kinases. In the present study, we have investigated the role of the PKC-alpha isoenzyme in FcgammaR-mediated phagocytosis using clones of the mouse macrophage cell line RAW 264. 7 overexpressing a dominant-negative (DN) mutant of PKC-alpha. Overexpression of DN PKC-alpha had no effect on the attachment of IgG-opsonized sheep red blood cells, but inhibited their internalization. Further analysis of the signaling events induced by IgG-opsonized sheep red blood cells revealed that whereas tyrosine phosphorylation of Syk was normal, phosphorylation of ERK 1/2 (p42/44) was impaired in DN PKC-alpha-overexpressing macrophages. These observations suggest a role for PKC-alpha in the regulation of FcgammaR-induced phagocytosis and signal transduction. PMID- 11027500 TI - Dual effect of insulin on in vitro leptin secretion by adipose tissue. AB - Although it is widely accepted that insulin stimulates leptin secretion, a dual action was observed using a validated in vitro system, i.e., an early (less than 48 h) inhibitory action, followed later (48-96 h) by a clear-cut stimulation. While the inhibitory phase was observed at every glucose concentration tested (from 1 to 25 mM), the stimulatory phase required the presence of physiological or supraphysiological glucose concentrations. In fact, leptin secretion was virtually eliminated in the presence of glucose uptake inhibitors. This dual effect of insulin was not due to modifications of the ob mRNA levels, suggesting that it depends entirely on posttranslational mechanisms. In conclusion, insulin appears to induce an early inhibition of leptin secretion by the adipose cell, followed later by a stimulatory effect secondary to the metabolic changes triggered by the insulin-induced increase in glucose uptake. PMID- 11027501 TI - A low-molecular-weight cytosolic inhibitor of the specific testosterone binding in bovine seminal vesicles. AB - Bovine seminal vesicle cytosol contains a low-molecular-weight and thermostable substance which specifically inhibits the binding of testosterone to its cognate receptor. The mass and the ether phospholipid structure of the inhibitor were elucidated by mass spectrometry. Saturation and binding experiments indicate that the inhibitor acts in a dose-dependent and competitive manner, altering the apparent dissociation constant (K(D)) while maintaining the number of androgen binding sites (B(max)). Its possible role in the regulation of androgen binding activity is discussed. PMID- 11027502 TI - Functional coupling of cyclooxygenase 1 and 2 to discrete prostanoid synthases in liver macrophages. AB - The profile of released prostanoids after addition of exogenous arachidonic acid to resident liver macrophages is different from the profile obtained in lipopolysaccharide-pretreated cells. In resident and lipopolysaccharide pretreated cells, AA leads to a release of thromboxane B(2), prostaglandin F(2alpha), E(2), and D(2). A specifically enhanced formation of prostaglandin E(2) is obtained in lipopolysaccharide-pretreated cells. Resident liver macrophages express cyclooxygenase 1, and thromboxane A(2)-, prostaglandin F(2alpha)-, E(2)-, and D(2)-synthase. Treatment with lipopolysaccharide induces in addition to cyclooxygenase 2-an enhanced expression of the prostaglandin E(2) synthase. In resident liver macrophages, the formation of prostanoids from exogenous arachidonic acid is completely inhibited by SC560 (a specific inhibitor of cyclooxygenase 1), but remains unchanged with SC236 (a specific inhibitor of cyclooxygenase 2). In lipopolysaccharide-pretreated liver macrophages, the formation of thromboxane B(2), prostaglandin F(2alpha) and D(2) is equally inhibited by SC560 and SC236 by about 50%. In contrast, the formation of prostaglandin E(2) is inhibited to a greater extent by SC560 (75%) compared to SC236 (26%). We conclude from these data, that in lipopolysaccharide-pretreated liver macrophages (i) cyclooxygenase 1 and 2 couple both to discrete prostanoid synthases, (ii) the functional coupling of cyclooxygenase 1 and 2 to the thromboxane A(2)-, prostaglandin F(2alpha)-, and D(2)-synthase is almost identical, and (iii) the enhanced prostaglandin E(2) synthesis is due to an enhanced expression of the prostaglandin E(2) synthase, which is coupled more efficiently to cyclooxygenase 1. PMID- 11027503 TI - Identification and functional characterization of the human and murine fibroblast growth factor receptor 4 promoters. AB - Fibroblast growth factor receptors (FGFRs) play crucial roles in signal transduction of adult tissues and during embryonic development. To study the transcriptional control, we isolated and characterized the promoter of human FGFR4. Two transcription initiation sites were identified. The deletion analysis in different cell types defined a core promoter reaching from -9 to -198, lacking TATA and CCAAT boxes but displaying high GC content (77%) in a stretch of 300 bp upstream of the major mRNA start. This region harbors multiple binding motifs for transcription factors. Moreover, the region between -1085 and -1140 contains a potential repressor element, which downregulates transcriptional activity. To identify conserved regulatory elements, we isolated and analyzed also the murine FGFR4 promoter. Only one transcription start was identified using RNase protection assays. Sequence alignment of human and mouse shows a striking similarity in the core promoter region of both genes, encompassing conserved transcription factor binding sites and a splice acceptor site. Furthermore, the region containing the putative repressor element is also conserved suggesting a functional role for gene expression. PMID- 11027504 TI - Phosphatidylinositol 3-kinase stimulates muscle differentiation by activating p38 mitogen-activated protein kinase. AB - The activation of both phosphatidylinositol 3-kinase (PI3-kinase) and p38 mitogen activated protein kinase (p38 MAPK) is required for muscle differentiation. However, it is not known whether the signals from these two kinases interact during this process. In this work, we have investigated this using H9c2 cardiac myoblasts. The p38 MAPK-specific inhibitor SB203580 blocked muscle differentiation and suppressed the expression of myogenin and myosin heavy chain in a concentration-dependent manner. Consistent with this, expression of a wild type p38 MAPK (Ha-p38) or a constitutively active MAPK kinase 6 (MKK6(glu)) promoted the rate of differentiation into multinucleated myotubes. LY294002, a PI3-kinase inhibitor, suppressed in a dose-dependent manner not only muscle differentiation but also activation of p38 MAPK. In addition, expression of a constitutively active form of PI3-kinase (p110*) enhanced myotube formation and p38 MAPK activation, while expression of a dominant negative form of PI3-kinase (Deltap85) attenuated these responses. Furthermore, SB203580 suppressed differentiation of H9c2 cells expressing p110*. Interestingly, LY294002 also suppressed differentiation of H9c2 cells expressing Ha-p38 or MKK6(glu). However, SB203580 did not affect PI3-kinase activity, suggesting that PI3-kinase myogenic signaling to p38 MAPK is unidirectional. Taken together, we concluded that PI3 kinase activates p38 MAPK, which in turn stimulates muscle differentiation, but that p38 MAPK does not substitute for PI3-kinase in this process. PMID- 11027505 TI - Heterogeneity at the 5'-end of MEN1 transcripts. AB - Multiple endocrine neoplasia type 1 is an autosomal dominant familial cancer syndrome. The responsible gene, MEN1, has been isolated and inactivating mutations have been found in the majority of MEN1 families. The underlying genetic defects in the remaining families may be located in yet unidentified regions of the MEN1 gene. Here, we present novel transcripts of MEN1 which vary in the content of their 5'-untranslated region. All transcript variants display upstream exons correctly spliced to MEN1 exon 2. The most commonly seen splice isoform occurred in a region previously published as human intron 1, a region which shows a high conservation between human and rodent MEN1. This splice variant uses an analogous transcription initiation site and identical splice donor/acceptor sites as a major transcript seen in rodent Men1. The newly identified MEN1 isoforms may represent biologically important transcripts and should thus be studied for mutations in the regions enclosed therein. PMID- 11027506 TI - Part of Xenopus translin is localized in the centrosomes during mitosis. AB - During oogenesis, maternal mRNAs are synthesised and stored in a translationally dormant form due to the presence of regulatory elements at the 3' untranslated regions (3'UTR). In Xenopus oocytes, several studies have described the presence of RNA-binding proteins capable to repress maternal-mRNA translation. The testis brain RNA-binding protein (TB-RBP/Translin) is a single-stranded DNA- and RNA binding protein which can bind the 3' UTR regions (Y and H elements) of stored mRNAs and can suppress in vitro translation of the mRNAs that contain these sequences. Here we report the cloning of the Xenopus homologue of the TB RBP/Translin protein (X-translin) as well as its expression, its localisation, and its biochemical association with the protein named Translin associated factor X (Trax) in Xenopus oocytes. The fact that this protein is highly present in the cytoplasm from stage VI oocytes until 48 h embryos and that it has been described as capable to inhibit paternal mRNA translation, indicates that it could play an important role in maternal mRNA translation control during Xenopus oogenesis and embryogenesis. Moreover, we investigated X-translin localisation during cell cycle in XTC cells. In interphase, although a weak and diffuse nuclear staining was observed, X-translin was mostly present in the cytoplasm where it exhibited a prominent granular staining. Interestingly, part of X-translin underwent a remarkable redistribution throughout mitosis and associated with centrosomes, which may suggest a new unknown role for this protein in cell cycle. PMID- 11027507 TI - Low calcium environment effects osteoprotegerin ligand/osteoclast differentiation factor. AB - In coculture with osteoblastic cell line MC3T3-E1 (E1) and mouse bone marrow cells, we reported that numbers of osteoclasts rose significantly on exposure to a low-calcium environment. Here we examined how osteoblasts influence osteoclastogenesis under a low-calcium environment. Comparing low extracellular calcium with a regular calcium environment, osteoprotegerin ligand (OPGL)/osteoclast differentiation factor (ODF) mRNA expression show more increase in the culture of low-calcium environment than in that of a regular calcium environment. Calcium-sensing receptor (CaSR), which was supposed as one of the mechanisms of recognizing extracellular calcium, existedon the surface of E1 cells. When E1 cells stimulated with agonists of CaSR, gadolinium, and neomycin, OPGL/ODF mRNA expression decreased. Moreover, these agonists reduced osteoclast formation in coculture. Taken together, it is possible that osteoblasts may recognize extracellular calcium via CaSR and regulate osteoclastogenesis. PMID- 11027508 TI - Characterization of human SCO1 and COX17 genes in mitochondrial cytochrome-c oxidase deficiency. AB - At least three proteins, COX17p, SCO1p, and its homologue SCO2p are thought to be involved in mitochondrial copper transport to cytochrome-c-oxidase (COX), the terminal enzyme of the respiratory chain. Recently, we and others have shown that mutations in SCO2 are associated with a lethal infantile hypertrophic cardiomyopathy (HCMP) with COX-deficiency. The majority of patients with a similar phenotype were, however, negative for SCO2 mutations, suggesting the other genes as candidates for this disorder. Here we report on the genomic organization of SCO1 and COX17 on human chromosomes 17 and 3 respectively, and the complete sequence analysis of COX17 and SCO1 in 30 patients with COX deficiency. Using a panel of human:mouse-monochromosomal hybrids, the expression of COX17 was specifically restricted to chromosome 3, indicating that the previously reported sequence on chromosome 13 represents a pseudogene. DNA sequence analysis of SCO1 and COX17 in nine patients with severe COX deficiency and fatal HCMP, and in 21 patients with other COX deficiency disorders, did not reveal any pathogenic mutations or polymorphisms. We conclude that neither SCO1 nor COX17 are common causes of COX deficiency disorders. PMID- 11027509 TI - Flavonoid baicalin inhibits HIV-1 infection at the level of viral entry. AB - Baicalin (BA) is a flavonoid compound purified from medicinal plant Scutellaria baicalensis Georgi and has been shown to possess anti-inflammatory and anti-HIV-1 activities. In an effort to elucidate the mechanism of the anti-inflammatory effect of BA, we recently found that this flavonoid compound was able to form complexes with selected chemokines and attenuated their capacity to bind and activate receptors on the cell surface. These observations prompted us to investigate whether BA could inhibit HIV-1 infection by interfering with viral entry, a process known to involve interaction between HIV-1 envelope proteins and the cellular CD4 and chemokine receptors. We found that BA at the noncytotoxic concentrations, inhibited both T cell tropic (X4) and monocyte tropic (R5) HIV-1 Env protein mediated fusion with cells expressing CD4/CXCR4 or CD4/CCR5. Furthermore, presence of BA at the initial stage of HIV-1 viral adsorption blocked the replication of HIV-1 early strong stop DNA in cells. Since BA did not inhibit binding of HIV-1 gp120 to CD4, we propose that BA may interfere with the interaction of HIV-1 Env with chemokine coreceptors and block HIV-1 entry of target cells. Therefore, BA can be used as a basis for developing novel anti-HIV 1 agents. PMID- 11027510 TI - Histamine H2 receptor mediated dual signaling: mapping of structural requirements using beta2 adrenergic chimeric receptors. AB - Previously we demonstrated that the histamine H2 receptor can activate both the adenylate cyclase and phosphoinositide/protein kinase (PKC) signaling pathways. Although dual coupling occurs via separate GTP-dependent mechanisms the structural components of the H2 receptor directing differential signaling have not been established. We explored this question by attempting to confer to the beta2-adrenergic receptor (betaAR), which is known to stimulate cAMP formation, the ability to activate PKC through the construction of beta2/H2 chimeric receptors. Intracytoplasmic domains of the human beta2 adrenergic receptor were substituted with the corresponding sequences of the human H2 receptor and stably expressed in HEK-293 cells. Binding of [(3)H]-CGP to chimeric wild type beta2 receptors was comparable. Substitution of the second intracellular loop (2i) of the betaAR led to a significant decrease in coupling to adenylate cyclase while leading to a 139.5 +/- 9.4% control increase in epinephrine mediated PKC activation. Introduction of the H2 receptor 3i also led to a decrease in betaAR mediated cAMP generation but provided the latter with the ability to stimulate PKC (182.2 +/- 8% of control). Concomitant expression of both 2i and 3i led to a substantial increase in epinephrine mediated PKC activation (201.8 +/- 10.5% of control). Addition of the carboxyl terminal tail did not facilitate stimulation of PKC. In summary, the third intracellular loop of the H2 receptor plays an essential role in activating PKC with maximal efficiency conferred by the second intracellular domain. PMID- 11027511 TI - Requirement of cell growth for gene expression induced by the lactose and tetracycline repressor-operator combination system in a human T cell line. AB - We applied the bacterial lactose and tetracycline repressor-operator systems to an interleukin 2-dependent T-cell line, Kit 225, to examine the effects of the human T-cell leukemia virus type I oncogene product, Tax, on the cell cycle. The LacSwitch and Tet-Off inducible systems individually exhibited low expression of Tax upon induction in growing Kit 225 cells. In contrast, combination of the LacSwitch system with the Tet-Off system produced a high Tax expression level in growing Kit 225 cells; however when arrested at the G0/G1 phase of the cell cycle, Kit 225 cells expressed very low levels of Tax, associated with little or no cell cycle progression. Infection with the Tax recombinant adenovirus induced high expression of Tax and progression of the cell cycle. Our results indicate that the combined LacSwitch and Tet-Off systems may require cell growth for gene expression. PMID- 11027512 TI - Alteration of rat dipeptidyl peptidase III by site-directed mutagenesis: cysteine(176) is a regulatory residue for the enzyme activity. AB - To comprehend the importance of cysteine residues for the regulation of enzyme activity, we replaced each of seven cysteine residues in rat dipeptidyl peptidase III cDNA with alanine, glycine, or glutamic acid residue using site-directed mutagenesis. Each mutated cDNA was expressed in E. coli (BL21), and each expressed DPP III was purified to apparent homogeneity on SDS-polyacrylamide gel electrophoresis. Six of the mutant proteins had similar activity to that of the wild-type enzyme, whereas the activity of the Cys(176) --> Ala mutant enzyme was only 25-35% of that of the wild-type enzyme activity. This mutant enzyme was resistant against both PCMB and NEM. Furthermore, both Cys(176) --> Gly and Cys(176) --> Glu mutated enzymes showed no DPP III activity. These seven mutated enzymes contained significant amounts of zinc, as determined by atomic absorption spectrometry. The results indicate that Cys(176) is essential for the regulation of DPP III activity. PMID- 11027513 TI - Self-amplification system for recombinant adeno-associated virus production. AB - A recently reported system for recombinant adeno-associated virus (rAAV) production does not require infection of a helper virus and depends on the transfection with a huge amount of three plasmids: AAV-vector, AAV-helper, and adenovirus-helper plasmids. Toward simplifying rAAV production, as a first step, we tested the use of the rAAV itself instead of the AAV-vector plasmid as a source of rAAV DNA and determined the optimal timing of infection and dose of the input rAAV. When 293 cells were infected just after transfection with 100 particles/cell of rAAV, irrespective of the purity, CsCl-purified or crude, up to 2000 particles/cell of rAAV were produced (9- to 20-fold self-amplification), a yield comparable to that obtained by an adenovirus-free transfection. These results indicate that infection of rAAV can greatly reduce the amount of plasmid DNA for a large-scale transfection. This strategy will also be useful when applied to packaging cell lines inducibly expressing Rep and Cap proteins. PMID- 11027514 TI - The new serine-threonine kinase, Qik, is a target of the Qin oncogene. AB - The winged helix transcription factor Qin is the avian homolog of the mammalian brain factor 1 (BF-1) and has the potential to act as an oncogenic protein. We used representational difference analysis to identify genes that are differentially expressed in chicken embryo fibroblasts (CEF) transformed by Qin. One of the up-regulated Qin targets identified in this analysis is a serine threonine kinase termed Qik (Qin-induced kinase). Qik belongs to the AMPK/SNF1 kinase family. It is a ubiquitously expressed protein and is upregulated rapidly after a hormone-regulated form of Qin is activated. In vitro kinase tests demonstrate that Qik is capable of autophosphorylation. Elevated levels of Qik transcripts are also observed in Src-transformed cells, suggesting that Src and Qin share some targets. PMID- 11027515 TI - Reactive oxygen intermediates are involved in IL-8 production induced by hyperosmotic stress in human bronchial epithelial cells. AB - Changes in the osmolarity of the airway surface fluid have been described to be involved in the pathogenesis of exercise induced asthma, and are suggested as the major cause of the lung disease in cystic fibrosis. In this study, we examined the signaling pathway of hyperosmotic challenge to interleukin-8 (IL-8). Hyperosmolarity (NaCl) caused a time- and concentration-dependent increase in IL 8 expression and secretion in bronchial epithelial cells. These effects could be blocked by antioxidants, such as DMSO, DMTU, DTT, and beta-mercaptoethanol, suggesting an involvement of reactive oxygen intermediates (ROI) in the signal transduction of hyperosmolarity-induced IL-8 synthesis. Since IL-8 is regulated by MAP kinases, we examined the influence of MAP kinase inhibitors on hyperosmolarity-induced IL-8 expression. The results show that this induction is regulated by p38 MAPK and not by ERK1/2. Furthermore, antioxidants blocked the activation of p38 MAPK induced by hyperosmolarity. These results suggest that ROIs are critical for p38 MAPK mediated IL-8 expression by hyperosmolarity. PMID- 11027516 TI - Transactivation of fibronectin promoter by HTLV-I Tax through NF-kappaB pathway. AB - The facts that fibronectin (FN) mRNA is elevated in cells expressing human T cell leukemia virus type I (HTLV-I) Tax protein and that Tax is known to transactivate the cellular cAMP-response element (CRE) prompted us to examine whether Tax activates the FN promoter of which CRE is thought to play an important role. We showed that Tax transactivated the FN promoter in Jurkat cells. Deletion analyses showed that the response-element resides within the promoter region of -69 bp and that an NF-kappaB-binding site at -41 bp is involved in the Tax-activation of the FN promoter. Gel-shift assays showed that DNA-protein complexes binding to the NF kappaB site, composed of NF-kappaB p50/p65, were induced on the NF-kappaB motif at -41 bp by Tax. Overexpression of NF-kappaB enhanced the Tax-activation of the FN promoter. Our study shows that the FN promoter is transactivated by Tax through the NF-kappaB pathway. PMID- 11027517 TI - Isolation of a novel ubiquitin-like protein from Pleurotus ostreatus mushroom with anti-human immunodeficiency virus, translation-inhibitory, and ribonuclease activities. AB - A glycoprotein, with a ubiquitin-like N-terminal sequence, has been prepared from an extract of fruiting bodies of the mushroom Pleurotus ostreatus, using a procedure which included ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose and Mono Q and gel filtration on Superdex 75. It exhibited a molecular weight of 12.5 kDa and was unadsorbed on DEAE-cellulose and Mono Q, but adsorbed on Affi blue gel and SP-Sepharose. It inhibited translation in a rabbit reticulocyte lysate system (IC(50) = 160 nM) and exhibited low ribonucleolytic activity (14 micro/mg) toward yeast tRNA. It also expressed an inhibitory activity toward human immunodeficiency virus-1 reverse transcriptase, which could be enhanced by succinylation. PMID- 11027518 TI - Cathepsin-B fusion proteins misroute secretory protein partners such as the proprotein convertase PC2-7B2 complex toward the lysosomal degradation pathways. AB - A general strategy is presented for the dominant negative reduction in the levels of heterodimeric soluble proteins within the secretory pathway through fusion of one of its partners C-terminal to the lysosomal enzyme cathepsin B (CB). Stable transfectants of CB-7B2 chimeras in AT20 cells result in a drastic reduction of the endogenous levels of its partner, the proprotein convertase PC2. This dominant negative suppressive effect requires active CB. It was partially reversed by NH(4)Cl, the cell-permeable CB inhibitor CA-074Me, but not by the proteasome inhibitor Lactacystin, suggesting the potential participation of the lysosomal/endosomal degradative pathway in this process. PMID- 11027519 TI - The PCNA from Thermococcus fumicolans functionally interacts with DNA polymerase delta. AB - We have cloned the gene encoding proliferating cell nuclear antigen (PCNA) from the hyperthermophilic euryarchaeote Thermococcus fumicolans (Tfu). Tfu PCNA contains 250 amino acids with a calculated M(r) of 28,000 and is 26% identical to human PCNA. Next, Tfu PCNA was overexpressed in Escherichia coli and it showed an apparent molecular mass of 33.5 kDa. The purified Tfu PCNA was tested first with recombinant Tfu DNA polymerase I (Tfu pol) and second with calf thymus DNA polymerase delta (pol delta). When tested with the homologous Tfu pol on bacteriophage lambda DNA, large amounts of Tfu PCNA were required to obtain two- to threefold stimulation. Surprisingly, however, Tfu PCNA was much more efficient than human PCNA in stimulating calf thymus pol delta. Our data suggest that PCNA has been functionally conserved not only within eukaryotes but also from hyperthermophilic euryarchaeotes to mammals. PMID- 11027520 TI - Imbalance production between interleukin-1beta (IL-1beta) and IL-1 receptor antagonist (IL-1Ra) in bronchial asthma. AB - Genes of the IL-1 family encode three different peptides, IL-1alpha, IL-1beta, and IL-1Ra, respectively. IL-1 operates through IL-1RI, and is involved in airway inflammation in asthmatic subjects, whereas IL-1Ra appears to be a specific competitive inhibitor of IL-1. All genes are on chromosome 2q12-21 where genomewide searches have identified linkage for asthma. To test whether variants of IL-1 relate to asthma, we conducted a genetic association study in a Japanese population. We show that the A2 allele of IL1RN (encoding IL-1Ra) associates with nonatopic asthma [OR = 5.71, 95% CI: 1.63-19. 8, Pc = 0.007]. Both atopic and nonatopic asthmatics with the A2 allele had significantly lower serum IL-1Ra levels in both types of asthmatics. Peripheral blood cells from asthmatics with A2 alleles, however, produced as much IL-1 as did those with A1 homozygotes. Since Th1 and Th2 cytokines differentially regulate the ratio between IL-1beta and IL-1Ra, these findings suggest that dysregulation of IL-1beta/IL-1Ra, probably due to interaction between epithelium and immuno-competent cells in the airway, is important in asthma inflammation. PMID- 11027521 TI - Hydrogen peroxide activation of Ca(2+)-independent phospholipase A(2) in uterine stromal cells. AB - In rat uterine stromal cells (U(III) cells), an oxidative stress induced by H(2)O(2) caused a dose-dependent release of arachidonic acid (AA) that was independent of intracellular Ca(2+) concentration and was not inhibited by Ca(2+) dependent phospholipase A(2) (cPLA(2)) inhibitors, nor by protein kinase C (PKC) inhibitors or by PKC down-regulation. H(2)O(2) treatment did not impair AA esterification but significantly increased Ca(2+)-independent PLA(2) (iPLA(2)) activity. Since iPLA(2) specific inhibitor bromoenollactone almost completely suppressed the release of AA induced by H(2)O(2), we conclude that iPLA(2) activity represents the major mechanism by which H(2)O(2) increases the availability of non-esterified AA in U(III) cells. Moreover, PKC inhibitors sphingosine and calphostin C markedly potentiated the release of AA trigger by H(2)O(2), suggesting a regulatory mechanism of iPLA(2) by PKC that remains to be clarified. PMID- 11027522 TI - Chaperone-like activity of bovine lens alpha-crystallin in the presence of dithiothreitol-destabilized proteins: characterization of the formed complexes. AB - Since alpha-crystallin was shown to have in vitro chaperone-like activity, its structure-function relationship has been extensively studied. However, the mechanism of this function is poorly understood. In this study, we monitored the interaction of alpha-crystallin with different proteins namely the insulin B chain (3.382 kDa), lysozyme (14.4 kDa), and conalbumin (86.18 kDa), all destabilized by dithiothreitol. We found that at 4 degrees C alpha-crystallin prevents the aggregation of destabilized insulin. During the time course of the interaction of alpha-crystallin with the substrate protein, we observed three classes of molecules: the monomeric protein and monomeric alpha-crystallin peptides, alpha-crystallin/substrate protein complexes with a size comparable to alpha-crystallin and large particles. The latter are responsible for the increase of the light scattering of solutions, containing alpha-crystallin/destabilized protein complexes. The molecular exchange between the different populations is temperature dependent and seems to be ruled by the kinetics of the structural changes of the destabilized proteins. At the end all monomeric species are transformed to larger aggregates. The large complexes are enriched in destabilized proteins, compared to the initial ratio alpha-crystallin/substrate protein. PMID- 11027523 TI - Doc2gamma, a third isoform of double C2 protein, lacking calcium-dependent phospholipid binding activity. AB - The Doc2 (double C2) family consists of two isoforms (Doc2alpha and Doc2beta) characterized by an N-terminal Munc13-1 interacting domain (Mid) and two C2 domains that interact with Ca(2+) and phospholipid at the C-terminus. This Ca(2+) binding property is thought to be important to the regulation of neurotransmitter release. In this paper, we report a third isoform of mouse Doc2, named Doc2gamma. Doc2gamma also contains a putative Mid domain and two C2 domains, and it is 45.6 and 43.2% identical to mouse Doc2alpha and Doc2beta, respectively, at the amino acid level. In contrast to the other Doc2 isoforms, the C2 domains of Doc2gamma impair Ca(2+)-dependent phospholipid binding activity. The highest expression of Doc2gamma mRNA was found in the heart, but occurs ubiquitously, the same as Doc2beta. These findings indicate that Doc2gamma may also function as an effector for Munc13-1 and that it may be involved in the regulation of vesicular trafficking. PMID- 11027524 TI - GTP cyclohydrolase I is coinduced in hepatocytes stimulated to produce nitric oxide. AB - GTP cyclohydrolase I is the rate-controlling enzyme in the production of tetrahydrobiopterin (BH(4)), an essential cofactor for nitric oxide (NO) synthase. Here we show that GTP cyclohydrolase I mRNA was present in unstimulated hepatocytes and was up-regulated 2- to 3-fold concurrently with iNOS induction induced in vivo by LPS injection and in vitro by stimulation with LPS and inflammatory cytokines tumor necrosis factor alpha, interleukin-1 beta, and interferon-gamma. Hepatocyte GTP cyclohydrolase I enzyme activity increased 2 fold in vivo after LPS. This coinduction of GTP cyclohydrolase I resulted in increased total intracellular biopterin which supported induced NO synthesis. The addition of a GTP cyclohydrolase I inhibitor to the stimulated hepatocytes decreased intracellular biopterin levels and resulted in a decrease in NO production. The results show that GTP cyclohydrolase I is up-regulated by certain acute inflammatory conditions. Further, the results indicate that biopterin is essential as a cofactor for induced NO synthase activity in hepatocytes. PMID- 11027525 TI - Chimeric proteins between UCP1 and UCP3: the middle third of UCP1 is necessary and sufficient for activation by fatty acids. AB - Uncoupling protein (UCP) 1 and UCP3 are mitochondrial inner membrane proteins which both mediate proton leak and thus decrease the mitochondrial transmembrane proton gradient. However, UCP1 and UCP3 differ in their biochemical regulation. UCP1 is activated by free fatty acids and inhibited by purine nucleotides. Using heterologous expression studies in yeast, UCP3 was found to lack both fatty acid activation and purine nucleotide inhibition. To assess which domains are responsible for the regulation of UCP1 by free fatty acids and by purine nucleotides and the absence of such regulation in UCP3, chimeric proteins were generated. Given that uncoupling proteins, like all members of the mitochondrial carrier family, possess a tripartite structure and consist of three repeated domains of approximately 100 residues, swaps in the three repeated domains were made between UCP1 and UCP3. Regulation of the resulting six different chimeric proteins by free fatty acids and purine nucleotides was studied after heterologous expression in yeast mitochondria. In this study, it is shown that activation of UCP1 by free fatty acids is mediated by the second repeated domain, since substitution of the second repeat of UCP1 by the equivalent repeat of UCP3 abolishes fatty acid activation. In contrast, replacing the second repeat of UCP3 by the corresponding repeated domain of UCP1 results in fatty acid activation similar to wild type UCP1. The lack of free fatty acid activation of UCP3 is not due to the absence of the histidine pair H145 and H147 found in the second repeated domain of UCP1. Furthermore, the findings with respect to purine nucleotide inhibition are consistent with a significant role of the C-terminal repeated domain of UCP1 in mediating purine nucleotide inhibition. PMID- 11027526 TI - Normal corpus callosum in Emx1 mutant mice with C57BL/6 background. AB - The Emx1 gene is a mouse homologue of a Drosophila head gap gene, empty spiracles. Corpus callosum defects have been reported for mutant mice lacking the Emx1 gene, indicating a possible involvement of the Emx1 gene in the regulation of axon guidance during development. However, it has recently come to light that genetic background could influence the outcome of the corpus callosum defects in mutant mice generated by ES cell technology. To investigate whether the corpus callosum defects resulted directly from Emx1 gene inactivation, we backcrossed the Emx1 mutant mice into the C57BL/6 background and found that the Emx1 homozygous mutant mice with the C57BL/6 background showed a normal corpus callosum score even though the cross-sectional area was 8% less than that of their wild-type littermates, indicating that genetic background could change the pattern of corpus callosum defects in Emx1 mutant mice. Furthermore, the indusium griseum and taenia tecta were always present and the cerebral cortical layers were well differentiated in Emx1 mutant mice. These results suggest that inactivation of the Emx1 gene does not contribute directly to the defects of corpus callosum and other brain structures associated with Emx1 mutant mice derived from a 129/Sv background. PMID- 11027527 TI - Tryptophan 67 in the human VPAC(1) receptor: crucial role for VIP binding. AB - The human receptor subtype for VIP and PACAP, referred to as VPAC(1) receptor, has a large N-terminal extracellular domain which is critical for VIP binding. We further investigated this domain by mutating 12 amino acid residues which could participate in the formation of a tight bend (W67) or a coiled coil motif. They were changed to alanine (A) and the cDNAs were transiently transfected into Cos cells. All mutants but W67A exhibited K(d) values similar to that of the wild type receptor. For the W67A mutant, no specific (125)I-VIP binding could be observed. Mutants at the W67 site were further characterized after stable transfection of epitope-tagged VPAC(1) receptor-GFP fusion proteins into CHO cells. W67A, W67E, W67H, and W67K mutants neither bound VIP nor mediated adenylyl cyclase activation by VIP. The W67F mutant mediated stimulation of adenylyl cyclase only at high VIP concentrations. Microscopic analysis and antibody binding experiments showed that all mutants were similarly expressed at the cell surface of CHO cells. Therefore tryptophan 67 in the human VPAC(1) receptor plays a crucial role in VIP binding due, in part, to its aromatic moiety. PMID- 11027528 TI - Identification of novel transcription factor-like gene from human intestinal cells. AB - Intestinal crypt epithelial T84 cells form luminal structures and differentiate to intestinal enterocyte-like cells in response to IMR-90 fibroblast-secreted transforming growth factor-beta when grown within three-dimensional collagen gel. In search of TGF-beta regulated genes involved in this differentiation process, we isolated a TGF-beta downregulated cDNA, human homologue of rat apoptosis antagonising transcription factor that codes for a 560-amino-acid protein. Human AATF-mRNA was expressed at high levels in human brain, heart, thymus, kidney, and placenta while in skeletal muscle and colon the expression was lower. The gene was mapped to chromosome 17q11.2-q12. PMID- 11027529 TI - Internalization and degradation of type IIA phospholipase A(2) in mast cells. AB - Whereas exogenous types IB and X secretory phospholipase A(2) (sPLA(2)) elicited prostaglandin D(2) (PGD(2)) production in mouse bone marrow-derived mast cells (BMMC), sPLA(2)-IIA was unable to do so. In search of a mechanism underlying this cellular refractoriness to exogenous sPLA(2)-IIA, we now report that this isozyme is promptly associated with cell surfaces, internalized, and then degraded in BMMC. Adsorption of sPLA(2)-IIA to BMMC was prevented by addition of heparin to the medium. Moreover, a heparin-nonbinding sPLA(2)-IIA mutant did not bind to BMMC. These results indicate that this sPLA(2)-IIA inactivation process depends on its rapid binding to heparan sulfate proteoglycan (HSPG) on BMMC surfaces. Thus, the present observations represent a particular situation in which cell surface HSPG exhibit a negative regulatory effect on cellular function of sPLA(2) IIA, and argue that HSPG does not always act as a functional adapter for heparin binding sPLA(2)s in mammalian cells as has been demonstrated before. PMID- 11027530 TI - Inhibition by deacetylase inhibitors of IL-1-dependent induction of haptoglobin involves CCAAT/Enhancer-binding protein isoforms in intestinal epithelial cells. AB - Intestinal epithelial cells participate in an acute phase response (APR) by responding to cytokines and by expressing acute phase protein genes. We hypothesized that butyrate, a fermentation product of the bacterial intestinal flora with deacetylase activity, affects the APR in intestinal epithelial cells. Sodium butyrate (NaBu) and Trichostatin A (TSA) induced alkaline phosphatase activity and histone H4 acetylation in IEC-6 rat intestinal epithelial cells treated with or without interleukin-1beta (IL-1). In contrast, both NaBu and TSA attenuated the IL-1-dependent induction of the acute phase protein gene haptoglobin, as well as C/EBPbeta and C/EBPdelta transcription factors mRNAs. Gel shift and supershift assays showed a strong decrease in the IL-1-induced C/EBPbeta and C/EBPdelta containing complexes binding to the HaptoA C/EBP DNA binding site of the haptoglobin promoter, by NaBu and TSA. Furthermore, site specific mutation of the HaptoA site abolished the NaBu- and TSA-dependent inhibition of haptoglobin, as determined by transient transfection assays. These results suggest that deacetylase inhibitors may regulate the IL-1 dependent induction of haptoglobin by down-regulating C/EBP isoforms, and that C/EBPs represent a target for the action of butyrate in the control of the APR of intestinal epithelial cells. PMID- 11027531 TI - Map kinase phosphatases (MKP's) are early responsive genes during induction of cerulein hyperstimulation pancreatitis. AB - Mitogen-activated protein kinase (MAPK) family members such as c-jun N-terminal kinase (JNK) may act as signal transducers early during pancreatitis development and evidence indicates that MAPK phosphatases (MKP) downregulate MAPK. We therefore investigated expression and regulation of pancreatic MKP in vivo. Pancreatic MKP mRNA levels were near or below the detection threshold in unstimulated animals. Cerulein hyperstimulation strongly induced MKP-1, MKP-3, and MKP-5 expression, peaking 30 to 60 min after treatment. Thus, MKP's clearly are early responsive genes during pancreatitis induction. Interestingly, inhibition of MKP-1 expression by Ro-31-8220 maximally induced activation of JNK but not of p38 and ERK in acutely isolated acini. These effects indicate that JNK may indeed be a preferred MKP-1 substrate in vivo. PMID- 11027532 TI - TGF-beta1 inhibits BRCA1 expression through a pathway that requires pRb. AB - TGF-beta1 inhibits BRCA1 expression, which contradicts the model that TGF-beta1 prevents carcinogenesis by activating tumor suppressor genes. To resolve this apparent contradiction, we examined BRCA1 expression in Mv1Lu cells, a well established model system for studying the TGF-beta1 tumor suppressor pathway. We found that inactivation of pRb by the papillomavirus type 16 E7 protein increased BRCA1 expression and abolished the ability of TGF-beta1 to inhibit BRCA1 expression. We conclude that TGF-beta1 inhibits BRCA1 expression through a pathway that requires pRb. We propose a model to explain the inhibition of BRCA1 as a target in the TGF-beta1 tumor suppressor signaling pathway. Our results suggest that the tumor suppressor functions of BRCA1 are initiated by the inactivation of pRb, and therefore that the activation of pRb by TGF-beta1 might alleviate the requirement for BRCA1 function. PMID- 11027533 TI - Pepstatin A-sensitive aspartic proteases in lysosome are involved in degradation of the invariant chain and antigen-processing in antigen presenting cells of mice infected with Leishmania major. AB - We previously reported that CA074, a specific inhibitor of cathepsin B, significantly deviated immune responses from the disease-promoting Th2 type to the protective Th1 type in BALB/c mice infected with Leishmania major. Herein, we found that pepstatin A-sensitive aspartic proteases (PSAP) in lysosomes seem to play a different role from that of cathepsin B in antigen-processing and Ii degradation. That is, cathepsin B appears to digest 16-, 28-, and 31-kDa peptides of soluble leishmania antigen (SLA), whereas PSAP seems to process mainly 28-kDa peptides. Furthermore, the latter protease contributed to the degradation of Ii but cathepsin B did not. Following treatment with pepstatin A, both Th1 and Th2 responses were profoundly suppressed in resistant DBA/2 mice (H-2(d)) and in susceptible BALB/c mice (H-2(d)), and both strains of mice became markedly susceptible compared with the untreated groups, probably owing to failure in degradation of Ii and partly to failure in digestion of 28-kDa peptide. PMID- 11027534 TI - Heat shock-induced, caspase-3-independent rapid breakdown of Akt and consequent alteration of its total phosphorylation/activity level. AB - The immediate effect of a 15-min heat shock was examined on the level and the activity of Akt. Following heat shock, the Akt level decreased by 15-70% in a temperature-dependent and phosphorylation status-independent manner. This decrease of Akt level was not prevented by caspase inhibitors. At 48 degrees C, the extent of the breakdown was so immense that the total phosphorylation/activity level of Akt was not increased over the control level, implying that the total cellular activity of Akt governed by the level and the molar activity does not necessarily undergo the ensuing change. PMID- 11027535 TI - Decreased locomotor activity in mice carrying transgenic Fyn tyrosine kinase. AB - We found that mice carrying constitutively active Fyn tyrosine kinase show low levels of spontaneous locomotor activity and that this activity increases when the mice are treated with the NMDA receptor antagonist MK-801. These findings indicate that the tyrosine phosphorylation of the NMDA receptors by Fyn participates in the control of locomotor activity. PMID- 11027536 TI - Transcriptional regulation of the human prepronociceptin gene. AB - We have cloned regions of the 5'-untranslated region of the human prepronociceptin (ppN/OFQ) gene into a luciferase reporter plasmid, pGL3 Basic Vector. By primer extension analysis, we determined that one start site of transcription lies within the known human cDNA sequence. There are two cyclic AMP response elements (CRE) with the consensus sequence CGTCA within 250 bp of the start of transcription. We designed various constructs around these sites, performed transient transfections, and measured luciferase activity in NS20Y cells. Transcriptional activity could be regulated by a variety of factors including cAMP levels, Ca(2+), and particularly by a 125-bp region adjacent to an intron located 23 bp upstream of the translation initiating ATG. These data should help in understanding the regulatory mechanisms of ppN/OFQ gene transcription. PMID- 11027537 TI - Oxidized LDL induces an oxidative stress and activates the tumor suppressor p53 in MRC5 human fibroblasts. AB - It is now well established that oxidized LDL (OxLDL) is involved in the progression of the atheromatous plaque via several mechanisms, including its cytotoxicity toward the arterial wall. Our study demonstrates that a 4-h incubation of cultured human fibroblasts with 25-75 microg/ml OxLDL induced a dose-dependent increase in the intracellular levels of reactive oxygen species (ROS) and lipid peroxidation end products (TBARS). This effect was markedly prevented by the antioxidant vitamin E. The lipid extract of OxLDL partially reproduced the action of the LDL particle itself. Concomitantly, OxLDL enhanced the DNA binding activity of p53 measured by electrophoretic mobility shift assay, and the intracellular protein level of p53 determined by immunoblot analysis. Cycloheximide prevented the OxLDL-induced augmentation in both p53 binding activity and intracellular level. Again, the lipid extract of OxLDL reproduced the effect of OxLDL on p53 binding activity, whereas vitamin E prevented it. These results indicate that OxLDL initiates an intracellular oxidative stress by means of its lipid peroxidation products, leading to the activation of the tumour suppressor p53 by enhancement of p53 protein synthesis. This effect might be related to the cytotoxic effect of OxLDL since the activation of p53 is known to lead to cell cycle arrest, necrosis or apoptosis. PMID- 11027538 TI - Expression of mPer1 and mPer2, two mammalian clock genes, in murine bone marrow. AB - Although circadian variations in hematopoiesis have been well documented, the molecular mechanism of the circadian rhythms remains elusive. To determine if a clock system exists in bone marrow to mediate the circadian rhythms, we analyzed the expression of mPer1 and mPer2, both mouse homologues of the Drosophila period gene and known regulators of the clock system, in murine bone marrow by relative quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). We demonstrated that both genes were expressed in bone marrow. Furthermore, the expression patterns of mPer1 and mPer2 in total bone marrow cells exhibited two peaks over a 24-h period. In contrast, the expression patterns of these two genes in the Gr-1-positive cells isolated from bone marrow mainly contributed to one of the two peaks. These results indicate that a clock system exists in bone marrow and suggest that the circadian rhythms in bone marrow are lineage- and/or differentiation stage-dependent. PMID- 11027539 TI - Low paraoxonase in Persian Gulf War Veterans self-reporting Gulf War Syndrome. AB - Exposure to organophosphate (OP's) insecticides and nerve gases during the Persian Gulf War has been implicated in the development of Gulf War Syndrome. Paraoxonase (PON1) present in human serum detoxifies OP's. We determined the levels of PON1 in the serum of Gulf War Veterans and compared these to those found in a control population. One hundred fifty-two Gulf War Veterans from the UK who self-reported the presence of Gulf War Syndrome via a questionnaire and 152 age and gender matched controls were studied. PON1 activity, concentration, and genotype were determined. In the Gulf War Veterans, paraoxon hydrolysis was less than 50% of that found in the controls (100.3 (14.8-233.8) vs 214.6 (50.3 516.2) nmol/min/ml, P < 0.001). This low activity was independent of the effect of PON1 genotype. The serum PON1 concentration was also lower in the Gulf War Veterans (75.7 (18.1-351.3) vs 88.2 (34.5-527.4) microg/ml, P < 0.00025), which was again independent of PON1 genotype. There was no difference in the rate of diazoxon hydrolysis between the groups (10. 2 +/- 4.1 micromol/min/ml vs 9.86 +/- 4.4, P = NS). A decreased capacity to detoxify OP insecticides resulting from low serum PON1 activity may have contributed to the development of Gulf War Syndrome. PMID- 11027540 TI - Cloning and characterization of the gene encoding the mouse peptide transporter PEPT2. AB - Here we describe the cDNA structure, genomic organization, chromosomal localization, and promoter analysis of the mouse peptide transporter PEPT2. The PEPT2-cDNA is 3987 bp long and encodes a protein of 729 amino acids. The functional properties, analyzed by expression in Xenopus laevis oocytes, showed a typical PEPT2-phenotype with electrogenic, proton-coupled transport, high substrate affinity, and a broad specificity. Immunoblotting of renal brush-border membranes revealed an apparent molecular mass of PEPT2 of 100 kDa. The murine Pept2 gene was cloned from a 129/SvevTACfBr genomic library. It is 34 kb long and comprises 22 exons and 21 introns. By radiation mapping analysis the Pept2 gene was mapped on central mouse chromosome 16. Two putative transcription start sites lying 35 and 235 bp upstream from the translation start were identified. The Pept2 gene possesses a TATA-less promoter. Functional promoter analysis revealed the core promoter to be located between 432 and 286 bp upstream from the translation start. PMID- 11027541 TI - Diphenyleneiodonium prevents reactive oxygen species generation, tyrosine phosphorylation, and histamine release in RBL-2H3 mast cells. AB - Mast cells play a central role in immediate allergic reactions mediated by immunoglobulin E. It has recently been reported that mast cells generate intracellular reactive oxygen species (ROS) in response to stimulation with divergent physiologically relevant stimulants. However, the physiological role of ROS is poorly understood. Here we demonstrate that mast cell model rat basophilic leukemia (RBL-2H3) cells generate ROS in response to antigen and the calcium ionophore A23187 via activation of diphenyleneiodonuim (DPI)-sensitive enzyme and that blockade of ROS generation by DPI suppresses histamine release induced by either stimulant. Increased tyrosine phosphorylation of pp125(FAK) and a 77-kDa protein coprecipitating specifically with the kinase occurred in parallel with the secretion, and blockade of ROS generation by DPI also suppressed the tyrosine phosphorylation of both proteins. These findings suggest that ROS generated by a flavoenzyme-dependent mechanism may be involved in histamine release through the pp125(FAK) pathway. PMID- 11027542 TI - Characterization of FcalphaR-triggered Ca(2+) signals: role in neutrophil NADPH oxidase activation. AB - Human neutrophil IgA receptors (FcalphaR) trigger phagocytosis of IgA-opsonized particles and activate the NADPH oxidase complex ultimately leading to pathogen destruction. Signal transduction events triggered by FcalphaR have not been investigated in the context of NADPH oxidase activation. In this study, we show that crosslinking FcalphaR triggers the release of Ca(2+) from an intracellular store that was unchanged by the addition of extracellular EGTA. This was in contrast to the thapsigargin-triggered Ca(2+) signal, which activates store operated Ca(2+) entry pathways (SOCP) and is sensitive to extracellular EGTA. Buffering extracellular Ca(2+) with EGTA had no effect on FcalphaR-triggered NADPH oxidase activation, suggesting that SOCP was not required for activation by FcalphaR. EGTA inhibited thapsigargin-triggered NADPH oxidase activation but had no effect on PMA-triggered responses. The intracellular Ca(2+) chelator BAPTA caused dose-dependent inhibition of both FcalphaR-triggered and thapsigargin triggered NADPH oxidase activation but had no effect on PMA-triggered responses. Our data demonstrate that FcalphaR-triggered NADPH oxidase activation is dependent on the release of Ca(2+) from an intracellular store, but is independent of SOCP. PMID- 11027543 TI - Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo. AB - Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70% at 1 microM concentration and its action, as demonstrated by a marked reduction (about 50%) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocal and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 microg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo. PMID- 11027545 TI - In vivo klotho gene delivery protects against endothelial dysfunction in multiple risk factor syndrome. AB - The klotho gene, originally identified by insertional mutagenesis in mice, suppresses multiple aging phenotypes (e.g., arteriosclerosis, pulmonary emphysema, osteoporosis, infertility, and short life span). We have previously shown that mice heterozygous for a defect in the klotho gene upon parabiosis with wild-type mice show improved endothelial function, suggesting that the klotho gene product protects against endothelial dysfunction. In the present study, using the Otsuka Long-Evans Tokushima Fatty (OLETF) rat which demonstrates multiple atherogenic risk factors (e.g., hypertension, obesity, severe hyperglycemia, and hypertriglyceridemia) and is thus considered an experimental animal model of atherosclerotic disease, we show that adenovirus-mediated klotho gene delivery can (1) ameliorate vascular endothelial dysfunction, (2) increase nitric oxide production, (3) reduce elevated blood pressure, and (4) prevent medial hypertrophy and perivascular fibrosis. Based on these findings, klotho gene delivery improves endothelial dysfunction through a pathway involving nitric oxide, and is involved in modulating vascular function (e.g., hypertension and vascular remodeling). Our findings establish the basis for the therapeutic potential of klotho gene delivery in atherosclerotic disease. PMID- 11027546 TI - Non-steady-state O(2) diffusion in metmyoglobin solutions studied in a diffusion chamber. AB - In this study, we studied the "passive" diffusion through myoglobin solutions by determining the oxygen diffusion coefficient (DO(2)) and the oxygen permeability (permeability O(2)) of metmyoglobin (metMb) solutions (3-33 g. 100 mL(-1)) at 25 degrees C. These oxygen diffusion parameters were determined in a diffusion chamber using a non-steady-state method and were also determined of albumin solutions (4-32 g. 100 mL(-1)) and distilled water for comparison. From these parameters, the oxygen solubility (alphaO(2)) could be calculated, because alphaO(2) = permeabilityO(2)/DO(2). Both DO(2) and permeabilityO(2) decreased with increasing metMb and albumin concentration. The values of DO(2), permeabilityO(2) and alphaO(2) of both metMb solutions and albumin solutions were comparable with literature values of methemoglobin (metHb) and serum protein solutions. The values of the metMb solutions can be used in following studies of facilitated oxygen diffusion through myoglobin solutions. PMID- 11027544 TI - EPR spectrometry of cytochrome P450 2B4: effects of mutations and substrate binding. AB - The EPR spectra of NH(2)-terminal-truncated P450 cytochrome 2B4 and of several active site mutants that were previously shown to be profoundly altered in catalytic properties were determined. From these spectra it was seen that the truncated P450 2B4, like the full length cytochrome, exists as the low spin ferric form, but upon mutation of threonine 302 to alanine approximately 40% of the cytochrome is present as the high spin ferric form (g approximately 8, 4, 2). A similar situation was observed in the double mutant E310L T302A, but not in the single mutant E301L. A rhombic high spin signal (g approximately 8, 4, 2) was observed when a substrate such as styrene, benzphetamine, or cyclohexane was added to the truncated cytochrome. Accompanying this change was the appearance of a signal at g = 1.98. Conversely, an axial high spin signal was observed (g approximately 6, 6, 2) when cyclohexanecarboxaldehyde or 3-phenylpropionaldehyde was added to the truncated P450 2B4. PMID- 11027547 TI - The rearrangement of the human alpha(1)-acid glycoprotein/orosomucoid gene: evidence for tandemly triplicated genes consisting of two AGP1 and one AGP2. AB - The human alpha(1)-acid glycoprotein (AGP) or orosomucoid (ORM) is controlled by the two tandemly arranged genes, AGP1 and AGP2. The further duplication of the AGP1 gene has been suggested by a few duplicated ORM1 locus haplotypes including ORM1*F1. S and ORM1*B9. S, detected by isoelectric focusing. To clarify the triplication of the AGP gene, 39 DNA samples from Japanese subjects were studied by the long-range PCR of intergenic regions. The analysis of PCR products showed that the tandemly triplicated genes, AGP1A-AGP1B-AGP2, occurred on about 20% of chromosomes. These composites were divided into ORM1A*F1-ORM1B*S-ORM2*M and ORM1A*B9-ORM1B*S-ORM2*M by allelic variations. Furthermore, the former was classified into a few haplotypes by three synonymous sequence variations, which might have arisen through gene conversion-like events. The recombination breakpoints existed between the 5' flanking region and intron 2 of the AGP1B gene. Thus, it is likely that the rearrangement of the AGP gene has often occurred. PMID- 11027548 TI - Autocrine regulation of proliferation and extracellular matrix homeostasis in human fibroblasts. AB - In the late stages of the tissue repair process, as well as during normal tissue turnover, tissue homeostasis may rely mostly on autocrine mechanisms. Accordingly, we have cultured normal human fibroblasts on plastic surfaces and within three-dimensional collagen gels in order to study, in this environment, the action of autologous medium conditioned by the same cells. We have observed that inside collagen gels the autologous medium strongly restrains cell proliferation, due to fibroblast-secreted growth factors, whose inhibitory effect can be annulled by suramin. Furthermore, concerning extracellular matrix formation, conditioned medium has no effect on novel collagen synthesis, while it up-regulates collagenase MMP-1 only in cultures on plastic. On the other hand, it strongly inhibits the secretion of the collagenase inhibitor TIMP-1, irrespective of the substratum. This effect is completely blocked by SB 203580, an inhibitor of the p38 MAP kinase. The above suggest the presence of an autoregulatory mechanism involved in tissue homeostasis. PMID- 11027549 TI - TNF-alpha stimulation of MCP-1 expression is mediated by the Akt/PKB signal transduction pathway in vascular endothelial cells. AB - MCP-1 is expressed in a variety of cell types including vascular endothelial cells following induction by different stimuli such as tumor necrosis factor (TNF)-alpha. Although TNF-alpha stimulates MCP-1 expression and secretion, the mechanism by which TNF-alpha stimulates expression of the MCP-1 gene is not known. In this study, we examine the involvement of the phosphatidylinositol-3-OH kinase (PI3-kinase)-Akt/PKB pathway. Exposure of human umbilical vein endothelial cells (HUVECs) to TNF-alpha elicited the rapid phosphorylation of Akt/PKB. In HUVECs, wortmannin, a PI3-kinase inhibitor, inhibits TNF-alpha-mediated MCP-1 secretion at a dose-dependent manner. Constitutively active form of Akt/PKB induces transcription of the MCP-1 gene, and cotransfection of dominant negative Akt/PKB suppressed the activation of the MCP-1 promoter induced by TNF-alpha. These findings show that Akt/PKB participates in the TNF-alpha induction of MCP-1 gene transcription in endothelial cells. PMID- 11027550 TI - Engineering of heterologous cytochrome P450 in Acinetobacter sp.: application for pollutant degradation. AB - Many organisms do not contain the necessary biochemical armoury to carry out the initial oxidative attack of many pollutant chemicals. In the present study, Acinetobacter sp. strain BD413 has been genetically engineered to express the cytochrome P450 xenobiotic-metabolising enzyme CYP105D1 from Streptomyces griseus that has in its repertoire a diverse array of organic pollutants. Further, it is shown that the transformed Acinetobacter calcoaceticus strain BD413 can grow on pollutants unlike control bacteria not expressing cytochrome P450 and that was reflected in release of radiolabel with growth on radiolabelled chlortoluron. We show that cytochrome P450 can enhance the biodegrading repertoire of A. calcoaceticus and discuss the application of such results to bioremediation strategies. PMID- 11027551 TI - Regulation of Id gene expression during embryonic stem cell-derived hematopoietic differentiation. AB - To elucidate the role of helix-loop-helix (HLH) Id proteins in hematopoietic differentiation, we used a model of embryonic stem (ES) cell differentiation in vitro which gives access not only to hematopoietic myeloid progenitor cells but also to the more primitive blast colony-forming cell (BL-CFC), the in vitro equivalent of the hemangioblast that gives rise to blast cell colonies in the presence of VEGF. We first demonstrated that ES cell-derived blast cell colonies could be used as a model to study hematopoietic differentiation and maturation. We next established the expression profile of Id genes in this model. Transcripts of the four Id genes were present in ES cells. Id1, Id3 and Id4 gene expression was down-regulated during the development of blast cell colonies while that of Id2 was maintained. Thus, Id1, Id3, and Id4 proteins are candidates for being negative regulators of hematopoiesis in the model of hematopoietic ES cell differentiation in vitro. PMID- 11027552 TI - A fundamental transcription factor for bone and cartilage. AB - Cbfa1-deficient mice were found to show a complete lack of bone formation owing to the maturational arrest of osteoblasts. Cbfa1 plays key roles in the determination of osteoblastic lineage from multipotential mesenchymal cells, their differentiation into mature osteoblasts, and transcriptional regulation of bone matrix-related genes. Cbfa1 positively regulates chondrocyte maturation and osteoclast differentiation and is required for vascular invasion into cartilage. Therefore, complete elucidation of the function of Cbfa1 and its signaling would be of great benefit in understanding skeletogenesis. PMID- 11027553 TI - Gene structure and promoter function of murine Munc18-2, a nonneuronal exocytic Sec1 homolog. AB - Sec1 family proteins are regulators of diverse exocytic processes, from yeast to man. Three mammalian homologues, Munc18-1, -2, and -3 have been described. We have studied the structure and expression of the mouse Munc18-2 gene. The Munc18 2 gene comprises 19 exons whose sizes range from 50 to 158 bp, with a total gene size of approximately 11 kb. A single transcript of 2.1 kb is expressed in multiple non-neuronal murine tissues. Munc18-2 has a striking resemblance to Munc18-1 in structure despite only 60% sequence identity, suggesting a recent gene duplication event. Analysis of the region upstream of the transcription start site shows that Munc18-2 has a TATA-less promoter, with a consensus initiator (Inr) sequence at the start of transcription, several Sp1 binding sites, and strong promoter activity in RBL-2H3 mast cells. The region from +5 to 430 is more active than +5 to -800, suggesting upstream repressor elements. PMID- 11027554 TI - The dNTPase enzyme activity is inhibited by nucleic acids and contains a heat insensitive component. AB - The dNTpase enzyme has previously been shown to specifically hydrolyse monodeoxyribonucleoside triphosphates (dNTPs). The remnant nucleotide resulting from this hydrolysis lacks the terminal phosphate and is covalently attached as part of a 3 kDa species, which we have termed the product nucleotide binding particle or "PNBP." PNBP is resistant to numerous nucleases and RNases, suggesting that it is not a nucleic acid polymer. Given that the exclusive specificity of dNTPase for dNTPs suggests some associative cellular role for the enzyme in polynucleotide maintenance, the interaction of dNTPase with various nucleic acids has now been examined. It is demonstrated that dNTPase activity is significantly inhibited by addition of single-stranded DNA or tRNA, but not rRNA. The data presented also suggest that thio-dATP can substitute for conventional phosphoester dATP in the enzymatic reaction. It is also demonstrated that the dNTPase enzyme comprises both heat/proteolysis/denaturant stable and heat/proteolysis/denaturant-sensitive components and we propose that this stable component may be the precursor to liganded PNBP. PMID- 11027555 TI - Midkine inhibits bradykinin-stimulated Ca(2+) signaling and nitric oxide production in endothelial cells. AB - Effects of the heparin-binding growth factor midkine (MK) were investigated on endothelial Ca(2+) signaling and nitric oxide (NO) production. Bradykinin (10 nM) and thapsigargin (1 microM) provoked large Ca(2+) influxes under fura-2/AM fluoroscopy. Pretreatment with human MK dose-dependently (1-500 ng/ml) inhibited the Ca(2+) response to bradykinin but not that to thapsigargin. Anti-MK antibody prevented this effect. In Ca(2+)-free medium, MK greatly inhibited intracellular Ca(2+) store release by bradykinin and not that by thapsigargin, which effect was prevented by the antibody. Bradykinin increased NO production by 6.7-fold, which was inhibited 6, 44, 79, and 90% by MK at 1, 10, 100, and 500 ng/ml, respectively. MK did not affect thapsigargin-induced NO production. Our data clearly indicate that MK inhibits bradykinin-induced Ca(2+) response and NO production from endothelial cells. PMID- 11027556 TI - Identification of retinoic acid-responsive elements on the HNF1alpha and HNF4alpha genes. AB - Hepatocyte nuclear factor 1alpha (HNF1alpha) and HNF4alpha are liver-selective transcription factors and are essential for hepatocyte differentiation. This study demonstrates that HNF1alpha as well as HNF4alpha genes contain a direct repeat with a space of one nucleotide (DR1)-retinoic acid (RA) response element that can be bound and regulated by RA and retinoid x receptor alpha (RXRalpha) complex. Transient transfection experiments showed that RA increased the promoter activity of the HNF1alpha and HNF4alpha genes in Hep3B cells. Overexpression of RXRalpha further enhanced the activities of both genes. Two putative RXRalpha binding sites on the HNF1alpha (-295 to -276) and HNF4alpha (-418 to -399) genes have been characterized. By transient transfection, both sites positively responded to RA, and overexpression of RXRalpha in Hep3B cells increased the regulatory effect. Gel mobility shift assay demonstrated that these two DR-1 sites could be bound by RXRalpha specifically. These data suggest that the differentiation effect of RA on hepatocyte may be due to direct interaction of RXRalpha with the RA-responsive elements on the HNF1alpha and HNF4alpha genes. PMID- 11027557 TI - Characterization of an apoptosis inhibitory domain at the C-termini of FE65-like protein. AB - TR2(L) is a 56-amino-acid polypeptide that has been shown to block TNF cytotoxicity. FE65-like (FE65L) proteins possess this conserved TR2(L) sequence at their C-termini, whereas variations in the sequences are found in the FE65 proteins. To further analyze the antiapoptotic function of TR2(L), here we utilized an isolated murine partial FE65L cDNA that encodes an N-terminal phosphotyrosine-binding domain (PTB) and the conserved C-terminal TR2(L) sequence. When L929 cells were stably transfected with the FE65L cDNA or its 3' end TR2(L) DNA sequence, these cells became resistant to TNF killing. Replacement of the N-terminal PTB domain with GFP failed to abolish the FE65L-mediated TNF resistance. Ablation of the C-terminal TR2(L) sequence through frame-shift mutation resulted in a complete loss of the FE65L function against TNF. Various protein kinase inhibitors, including lavendustin A, tyrphostin, H7, and staurosporine, which may affect the PTB domain function, could not abolish the FE65L-mediated TNF resistance. A prolonged exposure of L929 cells to these inhibitors for 24 h resulted in cell death, whereas FE65L significantly blocked the cell death. Polyclonal antibodies were generated against a synthetic peptide and shown to interact with a 38-kDa FE65L in L929 cells. Hyaluronidase downregulates the expression of FE65L gene and protein in L929 cells, and this correlates with its enhancement of TNF killing of these cells. Together, our data indicate that the TR2(L) amino acid sequence is an apoptosis-inhibitory domain commonly present in the FE65 and FE65-like family proteins. PMID- 11027558 TI - Nitric oxide synthase and postischemic liver injury. AB - The objective of this study was to determine what roles the endothelial cell and inducible isoforms of nitric oxide synthase (eNOS, iNOS) play in ischemia and reperfusion (I/R)-induced liver injury in vivo in mice genetically deficient in each isoform of NOS. We found that 45 min of partial (70%) liver ischemia and 5 h of reperfusion induced substantial liver injury as assessed by the release of large and significant amounts of the liver-specific enzyme alanine aminotransferase (ALT) into the serum of wild-type (wt) mice. The enhanced ALT levels were not due to increased recruitment of potentially damaging PMNs, which could mediate hepatocyte injury, as neither histopathological inspection nor quantitative MPO determinations revealed the presence of PMNs in the liver at this time point. In addition, we observed a significant enhancement in liver injury in eNOS-deficient but not iNOS-deficient mice subjected to liver I/R compared to postischemic wt mice. Taken together, these data suggest that eNOS- but not iNOS-derived NO plays an important role in limiting or downregulating I/R induced liver injury in vivo following 5 h of reperfusion. PMID- 11027559 TI - Baculovirus P35 protein does not inhibit caspase-9 in a cell-free system of apoptosis. AB - The P35 protein derived from the baculovirus Autographa californica NPV has been characterized as an inhibitor of apoptotic cell death in a great number of organisms and situations. This potential has been further mapped to the capacity of P35 to inhibit all caspases investigated. Here we show that P35 does not inhibit caspase-9 activity in a cell-free system of mammalian caspase activation. In cell extracts, cytochrome c addition led to the activation of caspase-9, -3 and -7. When cytosolic extract from cells expressing P35 was added, caspase-9 mediated maturation of caspase-3 proceeded normally but caspase-3-mediated further events were prevented, such as complete processing of caspase-3, processing of caspase-7 and the appearance of DEVD-cleaving activity. The P35 protein from Bombyx mori NPV, which has been reported to have a much weaker anti apoptosis activity in vivo, was found also to have significant caspase-3 inhibiting activity. These data suggest that P35 evolved specifically to inhibit effector rather than initiator caspases. PMID- 11027560 TI - Molecular cloning of a human, hemicholinium-3-sensitive choline transporter. AB - Under many physiological circumstances, Na(+)- and Cl(-)-dependent, hemicholinium 3 (HC-3)-sensitive, high-affinity choline uptake (HACU) in cholinergic neurons is thought to be rate-limiting in the biosynthesis of acetylcholine (ACh). Based on sequence information provided by the Human Genome Project and the recently reported rat CHT1 (rCHT1), we cloned a human CHT cDNA from spinal cord. The hCHT cDNA encodes a protein of 580 amino acids having 93% identity to rCHT1 and 51% identity to the Caenorhabditis elegans homolog CHO-1, and is distantly related to members of the Na(+)-coupled glucose transporter (SGLT) gene family of Na(+) coupled glucose (SGLT), nucleoside and iodide transporters. Northern blot analysis reveals the expression of a approximately 5 kb transcript in human brain regions rich in cholinergic neurons including the putamen, spinal cord, and medulla. Expression of hCHT cDNA in COS-7 cells results in saturable, Na(+)/Cl(-) dependent choline uptake (K(m) = 1.2 microM) in membrane vesicles and [(3)H] HC-3 binding (K(d) = 4 nM) in membrane fractions, consistent with characteristics reported in mammalian cholinergic neurons. Using radiation hybrid mapping techniques, we localized the hCHT gene to human chromosome 2q12. These studies elucidate the primary structure and chromosomal localization of hCHT and provide a basis for mechanistic analysis of HACU regulation and an investigation of the role of hCHT in disease states. PMID- 11027561 TI - Interferon-gamma directly inhibits TRANCE-induced osteoclastogenesis. AB - The immune system has profound effects on bone remodeling. IFN-gamma, a major product of immune cells, potently inhibits bone resorption, but its mechanism of action is unknown. We found in cultures of stroma-free mononuclear precursors that IFN-gamma strongly suppresses TRANCE/RANKL-induced osteoclast formation in a dose-dependent manner. This direct effect on osteoclast progenitors was not due to stimulation of NO production by IFN-gamma, as the NOS inhibitors 1400W and L NAME were unable to reverse the suppression. However, TGFbeta(1), which has opposing actions to IFN-gamma on diverse cellular functions, was able to antagonize the effect of IFN-gamma. This suggests that IFN-gamma prevents osteoclast formation by actively directing the differentiation of osteoclastic progenitors toward an alternative cytocidal lineage to the osteoclast. PMID- 11027562 TI - Junctional adhesion molecule (JAM) is phosphorylated by protein kinase C upon platelet activation. AB - Junctional adhesion molecule (JAM) is a member of the immunoglobulin superfamily (IgSF) expressed in tight junctions of epithelial cells and endothelial cells, and implicated in transendothelial migration of leukocytes. Recently, JAM is reported to be constitutively expressed on circulating monocytes, neutrophils, lymphocytes subsets, and platelets. However, the role of JAM is not known. Here, we examined how phosphorylaton of JAM is regulated upon platelet activation. Phosphorylation of JAM was induced by thrombin, collagen, but not by ADP. The phosphorylated amino acids were shown to be serine residues by phosphoamino acid analysis. Inhibition of JAM's phosphorylation by PKC inhibitors and Ca(++) chelator suggests the involvement of conventional types of PKCs. By in vitro kinase assays, we demonstrated that JAM could be directly phosphorylated by cPKCs. We also demonstrated phosphorylation of Ser 284, a putative PKC phosphorylation site, by immunoblotting with anti-phosphoserine-JAM antibody in thrombin-stimulated platelets. In addition to the phosphorylation, JAM seemed to form clusters at several sites of cell-cell contact in aggregated platelets by immunoelectron microscopic study. We speculate that JAM may be directly phosphorylated by cPKC(s)upon platelet activation and that the phosphorylationmight be involved in platelet activation. PMID- 11027563 TI - Expression of ZRC1 coding for suppressor of zinc toxicity is induced by zinc starvation stress in Zap1-dependent fashion in Saccharomyces cerevisiae. AB - The ZRC1 gene was cloned as a multicopy suppressor of zinc toxicity in Saccharomyces cerevisiae. Zrc1 belongs to CDF (cation diffusion facilitator) family. The transporters belonging to this family are thought to play an important role in metal detoxification. However, we found that cell growth of zrc1Delta mutant was lowered under the metal-limited conditions, which was restored by zinc. The Zap1 transcription factor is crucial for expression of several genes responsive to zinc-starvation stress. The expression of ZRC1 was induced in Zap1-dependent fashion when the intracellular zinc level was decreased and this induction was repressed by zinc. These results imply an important role of Zrc1 in the zinc-starvation stress. PMID- 11027564 TI - Expression of integrated hepatitis B virus X variants in human hepatocellular carcinomas and its significance. AB - Hepatitis B virus X protein (HBX) has been implicated in the transactivation of diverse cellular genes and possibly also the pathogenesis of human hepatocellular carcinoma (HCC). We report the characterization of HBX variants from HBV-related human hepatocellular carcinoma (HCC). These HBX variants were integrated into the host chromosomes and also expressed in the HCC tissues. In addition, we report a novel in vitro HBX activity assay based on color changes that were indicative of the beta-galactosidase enzyme activity. Conducted in wheat germ lysates, the transactivating function of either wild type or mutant HBX protein was measured through their interaction with the Early Growth Response factor 1 (Egr-1) that controls the beta-galactosidase gene. Further analysis of these HBX deletion mutants using this assay may shed new insights on the significance of various mutations occurring in HCC-associated HBX. PMID- 11027565 TI - The mitochondrial tricarboxylate carrier: unexpected increased activity in starved silver eels. AB - The tricarboxylate carrier was purified to homogeneity from liver mitochondria of European eel at the silver and the yellow stage and functionally reconstituted into liposomes. Unexpectedly, the molecular activity of the tricarboxylate carrier obtained from silver eel was about twofold higher than that of the same protein from yellow eel, although eels at the silver stage stop feeding. Parallel changes were found in the activities of the lipogenic enzymes in silver eels. This suggests a functional coordination between all these proteins sequentially involved in hepatic lipogenesis. Cardiolipin added to proteoliposomes strongly stimulated the activity of the purified tricarboxylate carrier from yellow eels, whereas it slightly reduced the activity of the same protein from silver eels. The higher activity of the tricarboxylate carrier from silver eels could therefore be ascribed, at least in part, to a different composition of the lipid domain surrounding the carrier protein, possibly in response to the hormonal alterations accompanying metamorphosis from yellow to silver stage. PMID- 11027566 TI - Sodium sulphate reactivates a protein A minidomain with a short elastin beta turn. AB - Elastin polymer sequences derived from muscle exhibit temperature and salt induced reversible contractions and expansions. A folded and contracted beta-turn helical elastin structure is stabilised by increased intramolecular hydrophobic interactions. We have generated a switchable binding protein by inserting a typical elastin turn sequence, GVPGVG, between the two IgG-binding helices of an engineered globular minidomain from Protein A. This mutant showed increased binding of IgG-molecules compared to the wild-type sequence in the presence of sodium sulphate, as measured by surface plasmon resonance spectroscopy. Analysis by circular dichroism revealed a salt-induced folding of the mutant minidomain to a native type I beta-turn, likely stabilised as a result of the two interacting dehydrated valines across the beta-turn. Since sodium sulphate can be successfully used to regulate the folding/unfolding or binding/dissociation of this minidomain, this suggests an alternative protein purification method. PMID- 11027567 TI - Ghrelin stimulates gastric acid secretion and motility in rats. AB - Ghrelin, a novel growth-hormone-releasing peptide, was discovered in rat and human stomach tissues. However, its physiological and pharmacological actions in the gastric function remain to be determined. Therefore, we studied the effects of rat ghrelin on gastric functions in urethane-anesthetized rats. Intravenous administrations of rat ghrelin at 0.8 to 20 microgram/kg dose-dependently increased not only gastric acid secretion measured by a lumen-perfused method, but also gastric motility measured by a miniature balloon method. The maximum response in gastric acid secretion was almost equipotent to that of histamine (3 mg/kg, i.v.). Moreover, these actions were abolished by pretreatment with either atropine (1 mg/kg, s.c.) or bilateral cervical vagotomy, but not by a histamine H(2)-receptor antagonist (famotidine, 1 mg/kg, s.c.). These results taken together suggest that ghrelin may play a physiological role in the vagal control of gastric function in rats. PMID- 11027568 TI - Cholesterol interaction with the daunorubicin binding site of P-glycoprotein. AB - The inherent complexities of cholesterol disposition and metabolism preclude a single transmembrane active transport avenue for this steroid-precursor, cell membrane constituent. Yet the ABC (ATP binding cassette) transporters are inextricably linked to elements of cholesterol disposition. Recent observations have suggested that, under certain settings, the ABC transporter P-glycoprotein (P-gp) performs a direct role in cholesterol disposition. The gene product of MDR1 (multidrug resistance transporter), P-glycoprotein also confers protection against xenobiotics. Using a whole cell assay in which the retention of a marker substrate is evaluated and quantified, we studied the ability of cholesterol to inhibit directly the function of this transporter. In a NIH-G185 cell line presenting an overexpressed amount of the human transporter P-gp, cholesterol caused dramatic inhibition of daunorubicin transport with an IC(50) of about 8 microM yet had no effect on the parent cell line nor rhodamine 123 transport. Additionally, using the ATP-hydrolysis assay, we showed that cholesterol increases P-gp-mediated ATP hydrolysis by approximately 1.6-fold with a K(s) of 5 microM. Suggesting that cholesterol directly interacts with the substrate binding site of P-gp, these results are consistent with cholesterol being transported by MDR1 P-gp. PMID- 11027569 TI - Correction of glycogen storage disease type II by enzyme replacement with a recombinant human acid maltase produced by over-expression in a CHO-DHFR(neg) cell line. AB - Inherited genetic deficiency of lysosomal acid alpha glucosidase or acid maltase (GAA) results in the autosomal recessive glycogen storage disease type II (GSD II). To investigate whether we could generate a functional recombinant human GAA (rhGAA) for enzyme replacement therapy, we subcloned the cDNAs for human GAA and mouse dihydrofolate reductase (DHFR) into DHFR(neg) Chinese hamster ovary cells and established a stable cotransformant that expressed rhGAA. We cultured the recombinant cells in media with progressively increasing concentrations of methotrexate and found that human GAA enzyme activity increased to over 2,000 IU per gram protein. Importantly, the human GAA enzyme activity correlated to equivalent amounts of human GAA protein by rocketimmunoelectrophoresis. We confirmed that the human GAA enzyme activity corresponded to an amplification in human GAA mRNA by Northern analysis and human GAA cDNA copy number by Southern analysis. Exposing the rhGAA to human GSDII fibroblast cells or patient's lymphocytes or monocytes resulted in uptake of the rhGAA and reversal of the enzymatic defect. Mannose-6-phosphate in the media blocked uptake. GAA -/- mice were treated with the rhGAA at 1 mg/kg, which resulted in heterozygous levels of GAA in tissues, most notably skeletal muscle, heart and diaphragm after two infusions. More importantly, after multiple infusions, hind, and fore-limb muscle weakness was reversed. This rhGAA would be ideal for enzyme replacement therapy in GSD II. PMID- 11027570 TI - The slow-binding inhibition of cathepsin K by its propeptide. AB - A peptide corresponding to the full-length proregion (amino acids 16-114) of human cathepsin K was expressed and purified from Escherichia coli. This recombinant propeptide was investigated for its ability to inhibit the activity of three cysteine proteinases: cathepsins K, L, and B. Kinetic studies showed the propeptide to be a potent slow-binding inhibitor of its parent enzyme with a K(i) = 2. 61 nM at pH 6. This inhibition was pH-dependent, with a decrease in pH from 6 to 4 leading to a concomitant increase in K(i) to 147 nM. The propeptide also inhibited cathepsin L with a K(i) = 26.1 nM at pH 6, but showed little inhibition of cathepsin B at concentrations up to 400 nM. PMID- 11027571 TI - Transforming growth factor beta inhibits the phosphorylation of pRB at multiple serine/threonine sites and differentially regulates the formation of pRB family E2F complexes in human myeloid leukemia cells. AB - Transforming growth factor beta (TGFbeta)1 induced dephosphorylation of pRb at multiple serine and threonine residues including Ser249/Thr252, Thr373, Ser780, and Ser807/811 in MV4-11 cells. Likewise, TGFbeta1 caused the dephosphorylation of p130, while inhibiting accumulation of p107 protein. Phosphorylated pRb was detected to bind E2F-1 and E2F-3, which appears to be a major form of pRb complexes in actively cycling cells. TGFbeta1 significantly downregulated pRb-E2F 1 and pRb-E2F-3 complexes as a result of inhibition of E2F-1 and E2F-3. In contrast, complexes of E2F-4 with pRb and with p130 were increased markedly upon TGFbeta1 treatment, whereas p107 associated E2F-4 was dramatically decreased. In agreement with these results, p130-E2F-4 DNA binding activity was dominant in TGFbeta1 treated cells, whereas p107-E2F-4 DNA binding activity was only found in proliferating cells. Our data strongly suggest that inhibition of E2Fs and differential regulation of pRb family-E2F-4 complexes are linked to TGFbeta1 induced growth inhibition. E2F-4 is switched from p107 to p130 and pRb when cells are arrested in G1 phase by TGFbeta1. PMID- 11027572 TI - Role of voltage-dependent anion channels in glutathione transport into yeast mitochondria. AB - Glutathione (GSH) is imported into mitochondria from the extra-mitochondrial cytoplasm. Translocation across the inner membrane of mitochondria is thought to occur via the dicarboxylate and 2-oxoglutarate carriers; however, the means by which GSH passes through the outer membrane is unknown. Disruption of the outer membrane of yeast mitochondria using either digitonin or osmotic shock did not alter GSH accumulation as compared with accumulation in intact mitochondria. These results suggested that passage across the outer membrane was not the rate limiting step in GSH accumulation. Mitochondria isolated from yeast strains with a disruption in the major pore-forming protein of the outer membrane, VDAC1, accumulated GSH to a greater extent than mitochondria isolated from a wild-type strain. Disruption of the gene for VDAC2 did not affect GSH import. Thus, neither VDAC form is essential for GSH translocation into mitochondria, and the participation of another outer membrane channel in GSH import is possible. PMID- 11027573 TI - Influence of oligomer chain length on the antioxidant activity of procyanidins. AB - The antioxidant activity of catechin monomers and procyanidin (dimers to hexamers) fractions purified from cocoa was studied in two in vitro systems: liposomes and human LDL. Liposome oxidation (evaluated as formation of 2 thiobarbituric acid reactive substances) was initiated with 2,2'-azobis (2 amidinopropane) hydrochloride (AAPH), 2,2'-azobis (2,4-dimethylvaleronitrile) (AMVN), iron/ascorbate, or UV-C; LDL oxidation (evaluated as formation of conjugated dienes) was initiated with Cu(2+) or AAPH. Catechin monomers and procyanidin fractions inhibited both liposome and LDL oxidation. Monomers, dimers, and trimers fractions were the most effective antioxidants when liposome oxidation was initiated in the aqueous phase. When oxidation was initiated in the lipid domains, higher molecular weight procyanidins were the most effective. All fractions significantly inhibited Cu-mediated LDL oxidation; no significant effect of procyanidin molecular weight was observed. The hexamer fraction was the least effective with respect to preventing AAPH initiated LDL oxidation. Results reported herein give further evidence on the influence of the oligomer chain length on the antioxidant protection by procyanidins. PMID- 11027574 TI - Brain-specific expression of novel G-protein-coupled receptors, with homologies to Xenopus PSP24 and human GPR45. AB - From mouse genomic libraries and human brain cDNA, we cloned three novel G protein-coupled receptors (GPCRs), which have about 55-70% homologies with Xenopus PSP24 (xPSP24). Together with another human cDNA (GPR45) cloned by Marchese et al. (Genomics 56, 12-21, 1999). they comprise a family of mammalian PSP24s. Therefore, we termed these clones mouse PSP24alpha, beta, and human PSP24alpha, beta. The homologies between alpha and beta isoforms were 54% for human and 51% for mouse clones. None of these clones shares sequence similarities with any known mammalian GPCRs, thus forming a unique gene family. Northern blot demonstrated that both of the mouse transcripts were predominantly expressed in the brain. In situ hybridization of brain sections showed that the expression was observed in neuronal cells, such as olfactory mitral cells, cortical neurons, hippocampal pyramidal cells, and Purkinje cells in the cerebellum. We suggest that mammalian PSP24 is a distinct GPCR family and plays a role in the brain function. PMID- 11027575 TI - Mammalian PSP24s (alpha and beta isoforms) are not responsive to lysophosphatidic acid in mammalian expression systems. AB - Xenopus PSP24 (xPSP24) is a G-protein-coupled receptor which was originally identified as a functional lysophosphatidic acid (LPA) receptor. We obtained two different types (alpha and beta) of mammalian homologues of xPSP24 and found that these receptors are highly expressed in the brain (Kawasawa et al., Biochem. Biophys. Res. Commun. 276, 952-956, 2000). These receptor did not respond to LPA by GTPgammaS binding assays, while Edg2 or Edg4 showed responses to LPA under the same assay conditions. Furthermore, a sensitive reporter gene assay using PC12 cells with serum response element promoter failed to detect any response of mammalian PSP24s to various concentrations of LPA. Thus, unlike xPSP24, we conclude that PSP24s are not functional LPA receptors in mammalian systems. PMID- 11027576 TI - Inhibition of N-linked glycosylation causes apoptosis in hamster BHK21 cells. AB - The tsBN7 cell line is one of the temperature-sensitive mutants for cell proliferation derived from hamster BHK21 cell line. It has a mutation in the DAD1 gene and enters apoptosis at the restrictive temperature of 39 degrees C. The defect of Dad1p causes a loss of N-linked glycosylation; therefore, it was thought that an inhibition of N-linked glycosylation induced apoptosis.However, tunicamycin, a potent inhibitor of N-linked glycosylation, had not caused apoptosis in wild-type BHK21 cells. In order to clarify this discrepancy, wild type BHK21 cells treated with tunicamycin and tsBN7 cells incubated at 39.5 degrees C were examined by the annexin V staining and TUNEL methods. Both methods showed that tunicamycin induces apoptosis in wild-type BHK21 cells, similar to the defect of Dad1p. Thus, we concluded that loss of N-linked glycosylation causes apoptosis. PMID- 11027577 TI - Sensitivity of duckweed (Lemna major) to ultraviolet-B radiation. AB - The sensitivity of an important aquatic macrophyte, duckweed (Lemna major), to UV B radiation was studied under experimental conditions at three different doses designated as no, mild, and severe injury dose by observing visible injury symptoms and estimating levels of chlorophyll, pheophytin, carotenoids, protein, starch, free sugar, and peroxidase activity. Laboratory-grown duckweed plants were exposed to UV-B radiation at 0.4 mW/cm(2) intensity for different time periods. Mild and severe injury were developed at 6.48 and 8.64 J, respectively. Peroxidase activity increased at all the exposure levels. Dose-dependent decrease in chlorophyll and starch with drastic depletion in protein and free sugar content were observed. Pheophytin and carotenoids content increased at no injury level, but decreased at higher exposure level. The results indicate that ambient UV-B radiation at the indicated level acts as a physiological stress in Lemna major. PMID- 11027578 TI - Isolation and characterization of murine orthologue of PTP-BK. AB - We isolated and characterized a murine orthologue of PTP-BK, a receptor-type protein tyrosine phosphatase expressed in brain and kidney. The deduced amino acid sequences showed 89, 95, and 92% identities with human, rabbit, and chicken homologues of PTP-BK. Mouse PTP-BK encodes a polypeptide of 1,226 amino acids with calculated molecular weight of 138,598 Da. The mature form of PTP-BK constitutes of 3 domain structures including extracellular, transmembrane, and a single intracellular PTPase domain. Western blot analysis indicated that anti-PTP BK antibody specifically immunoreacted to 2 distinct molecules of 200 kDa and 220 kDa in COS cells, possibly due to differential glycosylation. The recombinant PTP BK showed the phosphatase activity specific for the phosphotyrosine, but not for the phosphoserine residue of phosphopeptides in vitro. Radiation hybrid panel assigned mouse PTP-BK gene to 5.02cR distal to from the marker D6Mit339 on chromosome 6. Mouse PTP-BK was classified in PTPRO family in novel nomenclature. We discuss here the diversity and physiological functions of PTPRO family of PTP. PMID- 11027579 TI - The brain link protein-1 (BRAL1): cDNA cloning, genomic structure, and characterization as a novel link protein expressed in adult brain. AB - We report here molecular cloning and expression analysis of the gene for a novel human brain link protein-1 (BRAL1) which is predominantly expressed in brain. The predicted open reading frame of human brain link protein-1 encoded a polypeptide of 340 amino acids containing three protein modules, the immunoglobulin-like fold and proteoglycan tandem repeat 1 and 2 domains, with an estimated mass of 38 kDa. The brain link protein-1 mRNA was exclusively present in brain. When analyzed during mouse development, it was detected solely in the adult brain. Concomitant expression pattern of mRNAs for brain link protein-1 and various lectican proteoglycans in brain suggests a possibility that brain link protein-1 functions to stabilize the binding between hyaluronan and brevican. The human BRAL1 gene contained 7 exons and spanned approximately 6 kb. The entire immunoglobulin-like fold was encoded by a single exon and the proteoglycan tandem repeat 1 and 2 domains were encoded by a single and two exons, respectively. The deduced amino acid sequence of human brain link protein-1 exhibited 45% identity with human cartilage link protein-1 (CRTL1), previously reported as link protein to stabilize aggregates of aggrecan and hyaluronan in cartilage. These results suggest that brain link protein-1 may have distinct function from cartilage link protein-1 and play specific roles, especially in the adult brain. PMID- 11027580 TI - Cloning, expression, and mapping of hWW45, a novel human WW domain-containing gene. AB - WW domain-containing proteins are found in all eukaryotes and play an important role in the regulation of a wide variety of cellular functions such as protein degradation, transcription, and RNA splicing. The cloning and characterization of a novel human WW domain-containing gene, hWW45, which encodes a protein of approximately 45 kDa consisting of 2 WW domains and a coiled-coil region is reported here. The murine homologue cDNA, mWW45, displays a different 3' untranslated region and predicts a protein identity of 93% to hWW45. Northern blot and RT-PCR analysis demonstrated that both mWW45 and hWW45 transcripts are ubiquitously expressed in adult tissues. The mouse embryonic expression is first seen at 7 days post coitum in Northern blot analysis of whole embryos. Chromosomal localization by radiation hybrid mapping revealed that hWW45 is localized at chromosome 14, 10.31cR from the marker D14S269. PMID- 11027581 TI - A pivotal role of Zn-binding residues in the function of the copper chaperone for SOD1. AB - A Cu chaperone for SOD1 (CCS) is required for the incorporation of copper ion into the protein. To investigate the roles of the conserved metal-binding residues in CCS, we introduced amino acid substitutions into human CCS and examined the function of the mutant CCS by transforming a mutant yeast strain, SY2950, which lacks the lys7 gene, a yeast orthologue of the mammalian CCS. Mutant CCS in which amino acid residues His147 and Asp167 were substituted by Ala exhibited a decreased ability to complement the growth of SY2950 under Lys deficient conditions. This is because the mutations made the human CCS function in a less efficient manner, especially under metal-restricted conditions, leaving Cu,Zn-SOD in an apo-form. Since the His and Asp residues are both responsible for binding Zn which would serve to maintain the folded structure, the structural integrity supported by the coordinated Zn ion would be essential for CCS function. PMID- 11027582 TI - Stabilization of P450 2B4 by its association with P450 1A2 revealed by high pressure spectroscopy. AB - We studied the effect of intermolecular interactions between cytochromes P450 1A2 (CYP1A2) and 2B4 (CYP2B4) on the barotropic inactivation of the ferrous carbonyl complexes of the hemoproteins. When taken separately, these hemoproteins reveal quite distinct barotropic behavior. While the 2B4(Fe(2+))-CO complex is very sensitive to hydrostatic pressures and undergoes P450 --> P420 transition at rather low pressures (P(1/2) = 297 MPa, DeltaV(0) = -61 ml/mol), the 1A2(Fe(2+)) CO is extremely resistant to barotropic inactivation. Only about 8% of the 1A2 was exposed to pressure-induced P450 --> P420 transition (P(1/2) = 420 MPa, DeltaV(0) = -28 ml/mol). The formation of the mixed oligomers of 2B4 and 1A2 was found to have a dramatic effect on the barotropic behavior of 2B4. In the heterooligomers of 1A2 and 2B4, the 2B4 hemoprotein appears to be largely protected from barotropic inactivation. In 1:1 mixed oligomers no more than 25% of the total P450 content undergoes P450 --> P420 inactivation with the molar reaction volume value (DeltaV(0) = -26 ml/mol) similar to those found for pure 1A2. Moreover, interactions between 1A2 and 2B4 results in a displacement of the Soret band of the ferrous carbonyl complex of CYP2B4 to shorter wavelength (from 451.3 to 448.4 nm) and largely strengthens the dependence of the Soret band wavenumber on hydrostatic pressure below 200 MPa. This effect suggests an important hydration of the CYP2B4 heme moiety in response to the interactions with CYP1A2. We discuss these results in terms of the hypothesis that the heterooligomerization of cytochromes P450 in microsomes plays an important role in the control of the activity and coupling of the microsomal monooxygenase. PMID- 11027583 TI - The SepOvotropin: a new ovarian peptide regulating oocyte transport in Sepia officinalis. AB - In the cuttlefish Sepia officinalis, the successive steps of egg laying are controlled by multiple neuropeptides. Recent experiments led us to suppose that there was possible involvement of a second regulation pathway by the release of ovarian regulatory peptides in the genital tract. Using HPLC fractionation and an in vitro biological test, a C-terminal amidated peptide modulating the motility of the Sepia officinalis oviduct was isolated from an extract of vitellogenic ovarian follicles. The mass of this peptide as determined by MALDI-TOF (1501.8 Da) and analysis by Edman degradation led to the following sequence: Pro-Lys-Asp Ser-Met-Leu-Leu-Leu-Gln-Val-Pro-Val-Tyr-amide. The peptide mapping performed by LC/MS revealed a distribution restricted to the follicles, the full grown oocytes and the eggs. This new peptide, called SepOvotropin, modulated contractions of the whole genital tract in physiological conditions from a threshold concentration between 10(-20) and 10(-19) M, demonstrating for the first time the occurrence of a specific peptidergic control of egg-laying in cephalopods. PMID- 11027584 TI - Correlated gene expression between beta-1,4-galactosyltransferase V and N acetylglucosaminyltransferase V in human cancer cell lines. AB - Since our previous study showed that the gene expression level of beta-1,4 galactosyltransferase (beta-1,4-GalT) V is only increased in mouse NIH3T3 transformant and that beta-1,4-GalT V preferentially galactosylates the GlcNAcbeta1 --> 6Man branch of oligosaccharides [Shirane et al. (1999) Biochem. Biophys. Res. Commun. 265, 434-438], whether its gene expression is correlated with malignant transformation was investigated. Northern blot analysis of beta-1, 4-GalTs I, II, III, IV, V, and VI and N-acetylglucosaminyltransferase (GlcNAcT)V in human cancer cell lines showed that the gene expression levels of beta-1,4 GalT V but not other beta-1,4-GalTs are strongly correlated with those of GlcNAcT V whose activity was shown to increase by malignant transformation. These results indicate that beta-1,4-GalT V is involved in the galactosylation of highly branched oligosaccharides characteristic of malignantly transformed cells. PMID- 11027585 TI - PMCA1 mRNA expression in rat aortic myocytes: a real-time RT-PCR study. AB - The plasmalemmal Ca(2+) adenosine triphosphatase (PMCA) is a key regulator of Ca(2+) efflux in vascular smooth muscle. In these studies we developed a real time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) assay for assessing PMCA1 mRNA levels in rat primary cultured aortic myocytes. This assay detected fetal bovine serum-induced increases in PMCA1 mRNA (relative to 18S rRNA) 4, 8, and 24 h after stimulation. Early fetal bovine serum-induced increases in PMCA1 mRNA were insensitive to the Ca(2+) channel blockers nifedipine, flunarizine, and SKF-96365. These studies demonstrate the feasibility of real-time RT-PCR to assess mRNA levels of PMCA1 and illustrate dynamic regulation of this Ca(2+) pump isoform in rat primary cultured aortic myocytes. PMID- 11027586 TI - Functional analysis of human metaxin in mitochondrial protein import in cultured cells and its relationship with the Tom complex. AB - Metaxin is an outer membrane protein of mammalian mitochondria which is suggested to be involved in protein import into the organelle. RNA blot analysis showed that distribution of metaxin mRNA in human tissues differs from that of mRNA for the translocase component Tom20. Effect of overexpression of human metaxin on mitochondrial preprotein import and processing in COS-7 cells was studied. Overexpression of metaxin resulted in impaired mitochondrial import of natural and chimeric preproteins and in their accumulation. We previously reported that overexpression of Tom20 in cultured cells causes inhibition of import of mitochondrial preprotein. Coexpression of metaxin with Tom20 had no further effect on the preprotein import. Overexpression of the cytosolic domain of metaxin also caused inhibition of preprotein import, although less strongly than the full-length metaxin. In blue native PAGE, Tom40, Tom22, and a portion of Tom20 migrated as a complex of approximately 400 kDa, and the other portion of Tom20 migrated in smaller forms of approximately 100 and approximately 40 kDa. On the other hand, metaxin migrated at a position of approximately 50 kDa. These results confirm earlier in vitro results that metaxin participates in preprotein import into mammalian mitochondria, and indicates that it does not associate with the Tom complex. PMID- 11027588 TI - Identification of the functional site in the mosquito larvicidal binary toxin of Bacillus sphaericus 1593M by site-directed mutagenesis. AB - To study the mode of action of the binary toxin (51- and 42-kDa) of Bacillus sphaericus, amino acid residues were substituted at selected sites of the N- and C-terminal regions of both peptides. Bioassay results of the mutant binary toxins tested against mosquito larvae, Culex quinquefasciatus, revealed that most of the substitutions made on both peptides led to either decrease or total loss of the activity. Furthermore, receptor binding studies carried out for some of the mutants of the 42-kDa peptide showed mutations in N- and C-terminal regions of the 42-kDa peptide did not affect the binding of the binary toxin to brush border membrane vesicles of mosquito larvae. One of the mutants having a single amino acid substitution at the C-terminal region ((312)R) of the 42-kDa peptide completely abolished the biological activity, implicating the role of this residue in membrane pore formation. These results indicate the importance of the C-terminal region of the 42-kDa of binary toxin, in general, and particularly the residue (312)R for biological activity against mosquito larvae. PMID- 11027587 TI - Translocational status of ApoB in the presence of an inhibitor of microsomal triglyceride transfer protein. AB - Despite numerous studies demonstrating that microsomal triglyceride transfer protein (MTP) activity is critical to apoB secretion, there is still controversy as to whether MTP directly facilitates the translocation of apoB across the membrane of the endoplasmic reticulum (ER) through either the recruitment of lipids and/or chaperone activity. In the present study, a specific inhibitor of MTP (BMS 197636) was utilized in HepG2 cells to investigate whether a direct relationship exists between the translocation of apoB across the ER membrane and the lipid-transferring activity of MTP. Inhibition of MTP (with 10 and 50 nmol/L of the inhibitor) did not significantly affect the translocation of newly synthesized apoB (P = 0.77) or the translocational efficiency of the steady-state apoB mass (P = 0.45), despite a 49% decrease in apoB secretion and increased proteosomal degradation. These results compared well with subcellular fractionation experiments which showed no significant change in the fraction of apoB accumulated in the lumen of isolated microsomes in MTP-treated cells (P = 0.35). In summary, MTP lipid transfer activity does not appear to influence translocational status of apoB, but its inhibition is associated with an increased susceptibility to proteasome-mediated degradation and reduced assembly and secretion of apoB lipoprotein particles. PMID- 11027589 TI - Disease-associated mutations in SOD1 are impervious to dominant positive or negative effects. AB - The familial form of amyotrophic lateral sclerosis is caused by mutations in the SOD1 gene encoding the cytosolic antioxidant enzyme Cu,Zn superoxide dismutase. Although there is no clear correlation between disease and dismutating catalytic activity among the various disease-associated SOD1 alleles, all of the known missense mutations significantly alter the half-life of the encoded polypeptides. Using transient transfection studies in mammalian cells, it was demonstrated that a frameshift mutation in SOD1 which results in a truncated polypeptide is similarly destabilized. Using an epitope-tagging strategy to discriminate between mutant and wild-type SOD1 polypeptides, no evidence for dominant effects on polypeptide stability was detected, including that of a positive effect of the wild-type on mutant SOD1 polypeptides or that of a negative effect of mutant on wild-type SOD1 polypeptides. These experiments thus favor a non-catalytic role of mutant forms of SOD1 in disease progression. PMID- 11027590 TI - Gamma-glutamyltranspeptidase-dependent glutathione catabolism results in activation of NF-kB. AB - gamma-glutamyltranspeptidase (GGT) is a key enzyme implicated in the homeostasis of intracellular reduced glutathione (GSH) and hence in the regulation of the cellular redox state. Besides, the extracellular cleavage of GSH by GGT leads to reactive oxygen species (ROS) production, depending on the generation and enhanced reactivity of cysteinylglycine (CysGly). Using a model cell line, the V79 GGT, which highly expresses a human GGT transgene, we examined whether the GGT induced oxidant stress could modulate intracellular transcription factors. For the first time, we show that GGT-dependent ROS production induces the NF-kB binding and transactivation activities. This induction mimicked the one observed by H(2)O(2) and was inhibited by catalase, suggesting the involvement of H(2)O(2) in the NF-kB activation. PMID- 11027591 TI - Nuclear localization and apoptotic regulation of an amino-terminal domain focal adhesion kinase fragment in endothelial cells. AB - This study investigated the subcellular compartmentalization of focal adhesion kinase (FAK) fragments and their regulation during apoptosis of human umbilical vein endothelial cells. A 50 kDa NH(2)-terminal FAK fragment and a 120 kDa FAK variant were constitutively expressed and specifically found in the nuclear fraction of cells, while a 55 kDa COOH-terminal FAK fragment was only in the cytosolic fraction. FAK cleavage fragments generated during apoptosisremained in the cytosol, while p120FAK and p50 NH(2)-terminal FAK remained in the nuclear compartment. The caspase inhibitor, ZVAD-fmk, prevented the apoptosis-induced proteolysis of p125 and p120FAK, generation of the 80 kDa cleavage product, and increased expression of p50N-FAK. Western blot with phospho-specific FAK showed that nuclear p125(FAK) was phosphorylated at a significant level at Y861, while FAK phosphorylated at Y397 and Y407 was largely in the cytosol. These results indicate that FAK NH(2)- and COOH-terminal domain fragments are segregated between nuclear and cytosolic compartments in endothelial cells and suggest novel functions for the FAK NH(2)-terminal domain. PMID- 11027592 TI - Interaction of chloroquine and its analogues with heme: An isothermal titration calorimetric study. AB - Quinoline-containing drugs such as chloroquine and quinine have had a long and successful history in antimalarial chemotherapy. Identification of ferriprotoporphyrin IX ([Fe(III)PPIX], haematin) as the drug receptors for these antimalarials called for investigations of the binding affinity, mode of interaction, and the conditions affecting the interaction. The parameters obtained are significant in recent times with the emergence of chloroquine resistant strains of the malaria parasites. This has underlined the need to unravel the molecular mechanism of their action so as to meet the requirement of an alternative to the existing antimalarial drugs. The isothermal titration calorimetric studies on the interaction of chloroquine with haematin lead us to propose an altered mode of binding. The initial recognition is ionic in nature mediated by the propionyl group of haematin with the quaternary nitrogen on CQ. This ionic interaction induces a conformational change, such as to favour binding of subsequent CQ molecules. On the contrary, conditions emulating the cytosolic environment (pH 7.4 and 150 mM salt) reveal the hydrophobic force to be the sole contributor driving the interaction. Interaction of a carefully selected panel of quinoline antimalarial drugs with monomeric ferriprotoporphyrin IX has also been investigated at pH 5.6 mimicking the acidic environment prevalent in the food vacuoles of parasite, the center of drug activity, which are consistent with their antimalarial activity. PMID- 11027593 TI - The alpha-ketoisocaproate catabolism in human and rat livers. AB - Catabolism of alpha-ketoisocaproate in liver is mediated by cytosolic alpha ketoisocaproate dioxygenase (KICD) and mitochondrial branched-chain alpha-keto acid dehydrogenase complex (BCKDC). The latter is believed to be involved in the main pathway of the KIC catabolism. In the present study, we measured the activities of KICD and BCKDC in human and rat livers. The KICD activity in human liver was 0.9 mU/g tissue, which was 14.2% of the total activity of BCKDC, and that in rat liver was 4.2 mU/g tissue, which was only 1.0% of the total activity, suggesting that KICD in human liver plays a relatively important role in the alpha-ketoisocaproate catabolism. The KICD activity in human liver was significantly increased by cirrhosis. In rat liver, the enzyme activity was markedly increased by physical training and streptozotocin-induced diabetes, but not by feeding of a diet rich in branched-chain amino acids, although BCKDC activity was increased by feeding of the diet. PMID- 11027594 TI - Antigenicity of chimeric synthetic peptides based on HTLV-1 antigens and the impact of epitope orientation. AB - The present study evaluated four chimeric synthetic peptides incorporating immunodominant sequences from HTLV-1 virus. Monomeric peptides M1, M2, and M3 represent sequences from core (p19) and envelope (gp46) of the virus. The peptide M1 is a p19 (105-124) sequence, the peptide M2 is a gp46 (190-207) sequence, and the peptide M3 is a gp 46 sequence with substitution of proline at position 192 by serine. Those peptides were arranged in such a way that permits one to obtain different combinations of chimeric peptides (M1-M2, M2-M1, M1-M3, and M3-M1). Two glycine residues were used as arm spacers for separating the two sequences. The antigenicity of these peptides was evaluated in an ultramicroenzyme-linked immunosorbent assay (UMELISA) using sera of human T cell leukemia virus type I (HTLV-I)-infected individuals (n = 24), while specificity was evaluated with anti HTLV-II-positive samples (n = 11) and healthy blood donors (n = 25). The results were compared to plates coated with monomeric peptides M1, M2, and M3. The chimeric peptide orientation (M1-M2) and the proline at position 192 of the gp46 peptide showed higher sensitivity. PMID- 11027595 TI - Mouse flt-1 promoter directs endothelial-specific expression in the embyroid body model of embryogenesis. AB - The endothelial-specific receptor tyrosine kinase flt-1 (VEGFR-1) is expressed early on during endothelial lineage commitment both in vivo and in vitro. However, the exact function of flt-1 in vascular development still remains unclear. Here we report that a 2.2-kb fragment 5' of the mouse flt-1 gene becomes transcriptionally active during endothelial cell differentiation in developing embryoid bodies derived from mouse ES cells. Reporter gene expression correlated well with PECAM-1 expression and mirrored the expression pattern of the endogenous flt-1 gene. The temporal and spatial activity of the 2.2-kb flt-1 promoter provides a means to (1) identify a living population of early committed endothelial/bipotential progenitors and (2) ectopically express biologically active genes during lineage commitment. PMID- 11027596 TI - Upregulation of LOX-1 expression in aorta of hypercholesterolemic rabbits: modulation by losartan. AB - Angiotensin-II (Ang-II) enhances the modification of LDL and the expression of its lectin-like receptor (LOX-1) by activating type 1 (AT(1)) receptors. This study was designed to determine the effect of hypercholesterolemia on LOX-1 expression in aorta and its modulation by the AT(1) receptor blocker losartan. Male New Zealand White rabbits were fed regular chow (Control group), chow with 1% cholesterol and 4% peanut oil (HC-diet group), or 1% cholesterol and 4% peanut oil diet plus losartan (25 mg/kg/day) (Losartan + HC-diet group) for 10 weeks. Animal body weight, serum cholesterol levels, and arterial blood pressure were measured. Aortic intimal thickening was quantitated in H&E-stained segments. LOX 1 expression in aortas was examined by immunohistochemistry and semi-quantitative RT-PCR. High-cholesterol diet did not affect body weight, but induced hypercholesterolemia and extensive intimal thickening. Aortas of rabbits in the control group showed a modest LOX-1 expression in the endothelium. Aortic intimal proliferation in HC-diet group was associated with a marked increase in LOX-1 expression (protein and mRNA) in the endothelium and neointima. Treatment with losartan attenuated aortic intimal proliferation and markedly decreased the enhanced LOX-1 expression. Thus high-cholesterol diet induces the upregulation of LOX-1 expression in neointima of aortas of rabbits. Treatment with losartan, an AT(1) blocker, markedly decreases this enhanced LOX-1 expression. PMID- 11027597 TI - Ku70 can translocate to the nucleus independent of Ku80 translocation and DNA-PK autophosphorylation. AB - Ku plays an important role in multiple nuclear processes, e.g., DNA repair, chromosome maintenance, and transcriptional regulation. Although some evidence suggests that the nuclear translocation of Ku plays a key role in regulating the function of Ku, the mechanism is poorly understood. Using the site-directed mutagenesis technique, we demonstrate here that Ku70 can translocate to the nucleus without heterodimerization with Ku80. The nuclear accumulation of Ku70 mutants of the nuclear localization signal, which retained their binding ability with Ku80, was diminished. On the other hand, Ku70 mutants which lacked the ability to bind with Ku80 could translocate to the nuclei. Human Ku70, when transfected, accumulated within the nuclei of hamster xrs-6 cells which had undetectable DNA-PK activity and Ku80. Ku70 and Ku80 mutants of DNA-PK phosphorylation sites showed normal heterodimerization and nuclear translocation. These findings also support the idea that Ku70 can translocate to the nucleus independent of DNA-PK autophosphorylation. PMID- 11027598 TI - Localization of HB9 homeobox gene mRNA and protein during the early stages of chick feather development. AB - We performed in situ hybridization and immunohistochemical analysis of HB9 homeobox gene mRNA and protein, respectively, during chick feather development. HB9 mRNA was highly expressed in epidermal basal cells and dermal cells of the placodes and feather buds, but not in those of the interplacodes and interbud regions. HB9 protein was predominantly expressed in dermal cells of the symmetric short buds and decreased after the asymmetric bud stage when the feather bud had become elongated along the anterior-posterior (A-P) and proximal-distal (P-D) axis. These results suggest that HB9 gene is regulated in a spatiotemporal manner during feather development, and may be involved in early feather bud morphogenesis. PMID- 11027599 TI - Immunolocalization of AQP9 in liver, epididymis, testis, spleen, and brain. AB - The aims of this study were to determine the cellular and subcellular localization of aquaporin-9 (AQP9) in different rat organs by immunoblotting, immunohistochemistry and immunoelectron microscopy. To analyze this, we used rabbit antibodies to rat AQP9 raised against three different AQP9 peptides (amino acids 267-287, 274-295, and 278-295). In Cos7 cells transfected with rat AQP9, the affinity-purified antibodies exhibited marked labeling, whereas nontransfected cells and cells transfected with aquaporin-8 (AQP8) exhibited no labeling, indicating the specificity of the AQP9 antibodies. Immunoblotting revealed a predominant band of 28 kDa in membranes of total rat liver, epididymis, testes, spleen, and brain. Preabsorption with the immunizing peptides eliminated the labeling. Immunohistochemistry showed strong anti-AQP9 labeling in liver hepatocytes. The labeling was strongest at the sinusoidal surface, and there was little intracellular labeling. Immunoelectron microscopy revealed that the labeling was associated with the plasma membrane of the hepatocytes. In testes Leydig cells exhibited anti-AQP9 labeling, and in epididymis, the stereocilia of the ciliated cells (principal cells) exhibited significant labeling, whereas there was no labeling of the nonciliated cells (basal cells). This was confirmed by immunoelectron microscopy. In spleen strong labeling of cells was observed of leukocytes in the red pulp, whereas there was no labeling of cells in the white pulp. In rat brain, AQP9 immunolabeling was confined to ependymal cells lining the ventricles and to the tanycytes of the mediobasal hypothalamus. Antibody preabsorbed with the immunizing peptide revealed no labeling. In conclusion, AQP9 proteins is strongly expressed in rat liver, testes, epididymis, spleen, and brain. PMID- 11027600 TI - Myocilin expression in the astrocytes of the optic nerve head. AB - We investigated the expression of myocilin in the optic nerve head of porcine eyes by Western blotting and immunohistochemical staining. Myocilin was localized in the nucleus, centrosome, glial filament, mitochondria, and some parts of the cell membranes of the astrocytes. Myocilin was also detected at the edge-feet portion of the processes of astrocytes adjacent to the inner limiting membrane and blood vessel wall. The astrocytes are the major cell population in the optic nerve head, contributing to the architecture of the nerve axon and blood vessels. Therefore, myocilin gene mutation and change of myocilin protein are likely to affect the architecture of the optic nerve head and induce various forms of glaucomatous optic nerve damage. PMID- 11027601 TI - Expression and localization of heme oxygenase in human placental villi. AB - The aim of the present study was to investigate the expression and distribution patterns of heme oxygenase (HO)-1 and HO-2 in human placental villi at term and in the first trimester of pregnancy using reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemistry. RT-PCR detected mRNA for HO-1 and HO-2 in human placental villi during gestation. Western blotting also revealed the expression of the two distinct HO proteins throughout gestation. HO-1 was constitutively expressed, while HO-2 expression was apparently greater at term than in early pregnancy. Immunohistochemistry showed that distribution of the two HO isoforms had distinct topographic patterns: HO-1 was observed in villous trophoblastic cells, while HO-2 was found in endothelial cells and smooth muscle cells of blood vessels of placental villi. These results may provide a microtopographic basis for elucidating the mechanism of carbon monoxide (CO)-mediated vasodilatation, and it is suggested that the CO/HO system may be involved in the control of placental vascular function and may protect the syncytiotrophoblast and endothelium against oxidative injury. PMID- 11027602 TI - d-Aspartate in a prolactin-secreting clonal strain of rat pituitary tumor cells (GH(3)). AB - d-Aspartate (d-Asp) is found in prolactin (PRL)-containing cells of the rat anterior pituitary gland [Lee et al., Brain Res. 838, 193-199, 1999]. In order to determine whether d-Asp is actually produced by the anterior pituitary gland and whether it plays a physiological role in PRL function, a PRL-secreting clonal strain of rat pituitary tumor cells (GH(3)) was employed in this study. HPLC analysis and immunocytochemical staining detected the presence and synthesis of d Asp in the cytoplasm of these cells. In addition, thyrotropin-releasing hormone stimulated PRL secretion was increased in a dose-dependent fashion by d-Asp from these cells. These results suggest that the anterior pituitary gland synthesizes d-Asp and that d-Asp acts as a messenger in this gland. PMID- 11027603 TI - Solution structure of BmP01 from the venom of scorpion Buthus martensii Karsch. AB - From the venom of scorpion Buthus martensii Karsch,a short peptide (BmP01, 29 amino acid residues) was isolated and characterized as previously reported (Lebren, R. R., et al. (1997) Eur. J. Biochem. 245, 457-464). It was shown to reduce 33% outward K(+) channel (hippocampal neurons) currents at 10 microM. The solution structure of BmP01 was determined by 2D (1)H NMR spectroscopy. The NOEs, coupling constants, and H-D exchange obtained from NMR spectroscopy were used in structural calculations. The conformation of BmP01 is composed of a short alpha helix (Cys 3-Thr 12) and a two-stranded antiparallel beta-sheet (Ala 15-Asp 20 and Lys 23-Pro 28). There are three disulfide bridges (Cys 3-Cys 19, Cys 6-Cys 24 and Cys 10-Cys 26) connecting the alpha-helix and beta-sheet. Asp 20 to Lys 23 form a type II turn linking the two strands. Structural and electrostatic potential comparison between BmP01 and its analogues are also presented. PMID- 11027604 TI - Homeobox gene Hex is essential for onset of mouse embryonic liver development and differentiation of the monocyte lineage. AB - Disruption of the mouse Hex gene resulted in embryonic lethality around embryonic age (E) 10.5, due to no substantial liver formation. Expression of albumin was detectable in heterozygous (Hex(+/-)) but not in homozygous (Hex(-/-)) [corrected] embryos at E8.5. Instead of liver bud formation at E9.5, a liver-like capsule structure was observed in Hex(-/-) [corrected] embryos. In Hex(-/-) [corrected] mutant liver, we found no hepatocytes but no signs of apoptotic cell death in the area. Expression of transcription factors involved in hepatocyte differentiation, hepatocyte nuclear factor (Hnf)3beta, Hnf6, Hnf4alpha and Hnf1alpha, were restricted to the capsule and internal matrix-like structure in the mutant liver and expression of a subset of these factors were reduced. Hematopoiesis of monocytes was impaired in mutant embryos while erythroid lineage was unaffected. These results indicate that Hex plays an essential role in progenitor cells which commit to the hepatic endoderm and in the hematopoietic differentiation of the monocyte lineage. PMID- 11027605 TI - Domains of axin and disheveled required for interaction and function in wnt signaling. AB - Disheveled blocks the degradation of beta-catenin in response to Wnt signal by interacting with the scaffolding protein, Axin. To define this interaction in detail we undertook a mutational and binding analysis of the murine Axin and Disheveled proteins. The DIX domain of Axin was found to be important for association with Disheveled and two other regions of Axin (between residues 1-168 and 600-810) were identified that can promote the association of Axin and Disheveled. We found that the DIX domain of Disheveled is critical for association with Axin in vivo and for Disheveled activity. The Disheveled DIX domain controlled the ability of Disheveled to induce the accumulation of cytosolic beta-catenin whereas the PDZ domain was not essential to this function. PMID- 11027606 TI - Cloning and expression profiling of Hpa2, a novel mammalian heparanase family member. AB - Heparan sulfate proteoglycans are important constituents of the extracellular matrix and basement membrane. Cleavage of heparan sulfate by heparanase, an endoglycosidase, is implicated in the extravasation of leukocytes and metastatic tumour cells, identifying this enzyme(s) as a target for anti-inflammatory and anti-metastatic therapies. The cloning of a cDNA encoding human heparanase (Hpa1) was reported recently, together with evidence indicating that the hpa1 gene is unique and unlikely to belong to a family of related genes. Here we report the cloning of a cDNA encoding a novel human protein, HPA2, with significant homology to Hpa1. Alternative splicing of the hpa2 transcript yields three different mRNAs, encoding putative proteins of 480, 534, and 592 amino acids. Sequence analyses predict that all three Hpa2 proteins are intracellular, membrane-bound enzymes. Hpa2 also shows a markedly different mRNA distribution to Hpa1 in both normal and cancer tissues. The difference in expression profiles and predicted cellular locations suggests that Hpa2 and Hpa1 proteins have distinct biological functions. PMID- 11027607 TI - A Drosophila homolog of LIM-kinase phosphorylates cofilin and induces actin cytoskeletal reorganization. AB - Mammalian LIM-kinases (LIMKs) phosphorylate cofilin and induce actin cytoskeletal reorganization. To elucidate the functional roles of LIMKs in vivo during developmental processes, we attempted to isolate the cDNA encoding a Drosophila homolog of LIMK (DLIMK) and identified two isoforms of DLIMK transcripts coding for proteins with 1235 and 1257 amino acids, possessing the structure composed of two LIM domains, a PDZ domain, a protein kinase domain, and an unusual long C terminal extension. In situ hybridization analysis in Drosophila embryos detected the uniformly distributed DLIMK mRNA in stages 2 to 5. In vitro kinase reaction revealed that DLIMK efficiently phosphorylates Drosophila cofilin (twinstar) specifically at Ser-3, the site responsible for inactivation of its actin depolymerizing activity. When expressed in cultured cells, wild-type DLIMK, but not its kinase-inactive form, induced changes in actin cytoskeletal organization. These observations suggest that the LIMK-cofilin signaling pathway for regulating actin filament dynamics is evolutionarily conserved between Drosophila and mammals. PMID- 11027608 TI - Direct interaction of actin with p47(phox) of neutrophil NADPH oxidase. AB - The cell-free activation of human neutrophil NADPH oxidase is enhanced by actin, and actin filaments formed during activation are suggested to stabilize the oxidase. In an attempt to elucidate the mechanism, we examined the protein protein interactions between actin and cytosolic components of the oxidase. Far Western blotting using recombinant phox proteins showed that both alpha- and beta actin interacted with p47(phox) and rac1, and weakly with rac2. A deletion mutant of p47(phox) proved that its C-terminal region was essential for the interaction. The dissociation constant (K(d)) for interaction between actin and p47(phox) was estimated to be 0.45 microM by surface plasmon resonance, and that between actin and rac1 or rac2 was 1.7 or 4.6 microM, respectively. Far-Western blotting using cytosol as a target showed an interaction between actin and endogenous p47(phox) and rac proteins. These results suggest that actin can directly interact with p47(phox) and possibly with rac in the cells. PMID- 11027609 TI - Lateral clustering of cadherin-4 without homophilic interaction: possible involvement in the concentration process at cell-cell adhesion sites as well as in the cell adhesion activity. AB - It is thought that the concentration of classic cadherins at cell-cell adhesion sites is essential for generating strong cell-cell adhesion activity, but the mechanism is not well understood. To clarify the structural basis of the concentration process and the cell adhesion activity, we constructed various mutants of cadherin-4 and examined the adhesion properties of the transfectants. A deletion mutant lacking the entire cytoplasmic domain had weak, but significant Ca(2+)-dependent cell adhesion activity. Interestingly, the deletion mutant showed intrinsic cluster formation in the absence of cell-cell adhesion, possible lateral cluster formation. The cytoplasmic domain-deleted cadherin-4 containing the mutation of Trp-2 to Ala, which is known to inhibit the strand dimer formation required for the cell-cell adhesion, retained the possible activity of lateral cluster formation, supporting this notion. These results suggest that the extracellular domain has intrinsic activity of lateral cluster formation. Indeed, deletion of a cadherin repeat in the extracellular domain significantly reduced or abolished the lateral cluster formation as well as the concentration of cadherin-4 at cell-cell contact sites and cell adhesion activity. When transfectants of the cytoplasmic domain-deleted cadherin-4 made cell-cell contact and formed intimate cell-cell adhesion, the lateral clusters of cadherin-4 initially gathered at cell-cell contact sites, and a smooth linear concentration was gradually formed along the cell-cell adhesion interface. The results suggest that the lateral cluster formation is involved in the concentration process of cadherin-4 at cell-cell adhesion sites, hence in the strong cell adhesion activity of cadherin-4 as well. PMID- 11027610 TI - The H159A mutant of yeast enolase 1 has significant activity. AB - The function of His159 in the enolase mechanism is disputed. Recently, Vinarov and Nowak (Biochemistry (1999) 38, 12138-12149) prepared the H159A mutant of yeast enolase 1 and expressed this in Escherichia coli. They reported minimal (ca. 0.01% of the native value) activity, though the protein appeared to be correctly folded, according to its CD spectrum, tryptophan fluorescence, and binding of metal ion and substrate. We prepared H159A enolase using a multicopy plasmid and expressed the enzyme in yeast. Our preparations of H159A enolase have 0.2-0.4% of the native activity under standard assay conditions and are further activated by Mg(2+) concentrations above 1 mM to 1-1.5% of the native activity. Native enolase 1 (and enolase 2) are inhibited by such Mg(2+) concentrations. It is possible that His159 is necessary for correct folding of the enzyme and that expression in E. coli leads to largely misfolded protein. PMID- 11027611 TI - Molecular cloning and functional analysis of the promoter region of rat nonmuscle myosin heavy chain-B gene. AB - Rat nonmuscle myosin heavy chain-B (r-nmMHC-B) mRNA was previously found downregulated in Rat 6 fibroblasts transformed by mutant p53(val135) [J. W. P. Yam, J. Y. Zheng, and W. L. W. Hsiao (1987) Biochem. Biophys. Res. Commun. 266, 472-480]. Overexpression of exogenous r-nmMHC-B could partially reverse the transforming phenotypes both in vitro and in vivo. The downregulation of r-nmMHC B was also observed in Rat 6 transformed by c-H-ras and v-myc oncogenes. We cloned a 5.2-kb r-nmMHC-B promoter region. Sequence analysis of -1248 to +1 revealed no TATA box, but did show that it contained CAAT boxes, E12/E47, MyoD, MEF, E2F, CREB, and SP1 binding sites. Based on transient reporter assays, the promoter/enhancer activities were unusually extended to the entire 5.2 kb region in normal Rat 6 cultures, but markedly suppressed in p53(val135)-, and c-H-ras transformed cells. The activity detected by the reporter assay corresponded to levels of mRNA as analyzed previously by Northern blots in each respective cell line. Thus, the switch-off of the r-nmMHC-B in the transformed cells is very likely controlled by upstream transcriptional factors, which might have been altered in the course of neoplastic transformation. PMID- 11027612 TI - Ganglioside GD3 and its mimetics induce cytochrome c release from mitochondria. AB - Ganglioside GD3 induced the release of cytochrome c from isolated rat liver mitochondria. This process was completely prevented by cyclosporin A and partially prevented by a cysteine protease inhibitor, n-acetyl-leu-leu norleucinal. Cyclosporin A is a potent inhibitor of the permeability transition pore, whereas n-acetyl-leu-leu-norleucinal has no effect on this pore. These results indicate that the release of cytochrome c from mitochondria requires both the opening of the permeability transition pore and a cysteine protease inhibitor sensitive mechanism. Gangliosides GD1a, GD1b, GT1b, and GQ1b along with the synthetic GD3 mimetics TMS-42 and CI-22, which are glycerophospholipids carrying a disialo residue, also induced cytochrome c release. In contrast, gangliosides GM1, GM2, and GM3 did not induce cytochrome c release. These results indicate that two sialo residues must play an important role in the induction of cytochrome c release by gangliosides. PMID- 11027613 TI - Copper refolding of prion protein. AB - We have shown previously that normal mouse prion protein (MoPrP) binds copper ions during protein refolding and acquires antioxidant activity. In this report, we probe the structure of the copper refolded form of MoPrP to determine how copper binding alters the secondary and tertiary features of the protein. Circular dichroism showed that recombinant MoPrP prepared in the presence of copper (as Cu(++)) showed an increased signal in the 210-220 nm range of the spectrum. Changes in protein conformation were localised to the N-terminal region of MoPrP using a panel of antibodies to assess epitope accessibility. The copper refolded recombinant prion protein had reduced proteinase K (PK) sensitivity when compared to the non-copper liganded form. Reduced PK sensitivity was not due to aggregation however as high resolution electron microscopy showed a homogenous preparation with little aggregate when compared to the non-copper form. Finally, disruption of the single disulphide linkage in MoPrP significantly diminished the antioxidant activity of the copper refolded form suggesting that activity was not solely dependent on bound copper but also on a conformation enabled by the formation of the disulphide bond. PMID- 11027614 TI - Binding of tropomyosin-troponin to actin increases filament bending stiffness. AB - Rheologic measurements show that the association of tropomyosin-troponin with actin filaments is responsible for the reduction of the internal chain dynamic and increase in the mechanical rigidity of actin filaments. Basing calculations on the linear relation between the plateau modulus, G(N)('), and bending modulus, kappa, I find that tropomyosin-troponin at r(AT) = 7 increases actin filament stiffness by approximately 50%. This is confirmed by dynamic light scattering. Further increases are observed at rising F-actin and constant tropomyosin troponin concentrations. Tropomyosin-troponin also delays actin assembly and subsequent network formation and increases filament stiffness over time. PMID- 11027615 TI - The amino acid region 115-119 of ammodytoxins plays an important role in neurotoxicity. AB - Quadruple (Y115K/I116K/R118M/N119L) and double (Y115K/I116K) mutants of ammodytoxin A, a presynaptically toxic phospholipase A(2) from Vipera ammodytes ammodytes venom, were prepared and characterized. The enzymatic activity of the quadruple mutant on phosphatidylcholine micelles was threefold higher than that of AtxA, presumably due to higher phospholipid-binding affinity, whereas the activity of the double mutant was twofold lower. The substantial decrease by more than two orders of magnitude in the lethal potency of both mutants, together with their decreased binding affinity for neuronal receptors, indicates involvement of the amino acid region 115-119 in neurotoxicity. The similar decrease of toxicity for the two mutants points to the importance of the residues Y115 and I116. PMID- 11027616 TI - Asymmetric preference of serine proteases toward phosphonate and phosphinate esters. AB - We have previously reported the asymmetric synthesis of (alpha-aminoalkyl) diphenylphosphonate and phosphinate derivatives designed as inhibitors of chymotrypsin- and elastase-like proteases. This paper reports the first kinetic evaluation of individual epimers of the (alpha-aminoalkyl) diphenylphosphonates as inactivators of chymotrypsin, cathepsin G and neutrophil elastase (HNE). Results show that the (R)-epimers consistently function as more potent irreversible inactivators of their respective target proteases than the corresponding (S)-epimers. Additionally, phosphinate analogues were found to be consistently superior to their diphenylphosphonate counterparts. For example, Cbz. Phe(P)(OPh)-(CH(2))(2)-CO(2)Et inactivates cathepsin G approximately 45-fold more rapidly (k(i)/K(i) = 1.2 x 10(5) M(-1). min(-1)) than the analogous Cbz.Phe(P)(OPh)(2) (2.6 x 10(3) M(-1). min(-1)). Similarly, Cbz.Val(P)(OPh) (CH(2))(2)-CO(2)Et was found to inactivate HNE some 3-fold more efficiently than Cbz.Val(P)(OPh)(2) (6.5 x 10(3) and 2.0 x 10(3) M(-1). min(-1), respectively). PMID- 11027617 TI - The glycosyl phosphatidylinositol anchor of human T-cadherin binds lipoproteins. AB - T-cadherin (T-cad) is a Ca(2+)-dependent cell adhesion glycoprotein bound to the plasma membrane via a glycosylphosphatidylinositol (GPI) anchor. T-cad expressed on vascular smooth muscle cells (SMC) binds lipoproteins on blot. To analyze the molecular basis for the interaction of T-cad with lipoproteins we expressed recombinant human T-cad in HEK293 cells. Whereas membrane-bound T-cad from SMC and T-cad transfected HEK293 cells bind lipoproteins, T-cadherin proteins cleaved from the cell surface by phosphatidylinositol-specific phospholipase C (PI-PLC) do not. The lipoprotein-binding function is also lacking both for a recombinant human T-cad expressed in HEK293 cells without the GPI signal sequence, and for a human T-cad form expressed in Escherichia coli that contains the signal sequence for GPI attachment but is not modified with a GPI. We conclude that the GPI moiety of T-cadherin is necessary and sufficient to mediate lipoprotein binding. PMID- 11027618 TI - Identification of nuclear export signal in UL37 protein of herpes simplex virus type 2. AB - The UL37 gene of herpes simplex virus (HSV) encodes a 120-kDa phosphoprotein associated with the virion. In this study, we have generated a rabbit polyclonal antiserum against HSV-2 UL37 protein, and examined its intracellular localization by immunofluorescence study. In infected cells, specific fluorescence was detectable in the perinuclear region. In transfected cells, UL37 protein was observed mainly in the cytoplasm. Transfection assays of deletion mutants of UL37 protein suggested that the leucine rich region (LRR) containing amino acids 263 273 may be important for cytoplasmic localization. Deletion of the LRR or substitution of the leucine residues resulted in nuclear remaining of UL37 protein. Moreover, the LRR could export green fluorescent protein (GFP) to the cytoplasm as a fusion protein and this export was blocked by leptomycin B treatment, indicating that the LRR acted as a nuclear export signal. These results suggest that UL37 protein fulfills a role as a shuttle between the nucleus and the cytoplasm through the LRR. PMID- 11027619 TI - The ubiquitin-proteasome proteolytic pathway in heart vs skeletal muscle: effects of acute diabetes. AB - The ubiquitin-proteasome system is thought to play a major role in normal muscle protein turnover and to contribute to diabetes-induced protein wasting in skeletal muscle. However, its importance in cardiac muscle is not clear. We measured heart muscle mRNA for ubiquitin and for the C2 and C8 proteasomal subunits, the amount of free ubiquitin and the proteasome chymotrypsin-like proteolytic activity in control and diabetic rats. Results were compared to those in skeletal muscle (rectus). Heart ubiquitin, C2 and C8 subunit mRNA and proteolytic activity were significantly greater than in skeletal muscle (P 12 kb in size and contains 15 exons and 14 introns. The last coding exon (exon 15) contains a stop codon, leaving no room for PAL and, thereby, for a bifunctional PAM enzyme as in mammals. Furthermore, we found a CP1 splicing variant (CP1-B) that contains exon-9 instead of exon-8, which was present in the previously characterized PHM cDNA (CP1-A). CP1-A and -B have 97% amino acid sequence identity, whereas both splicing variants have around 42% sequence identity with the PHM part of rat PAM. Essential amino acid residues for the catalytic activity and the 3D structure of PHM are conserved between CP1-A, B and the PHM part of rat PAM. Furthermore, eight introns in CP1 occur in the same positions and have the same intron phasing as eight introns in the rat PAM gene, showing that the sea anemone PHM is not only structurally, but also evolutionarily related to the PHM part of rat PAM. PMID- 11027632 TI - Influence of reverse micellar environments on the fluorescence emission properties of tryptophan octyl ester. AB - A number of recent studies have presented perspectives on the hydrophobic fluorescence probe tryptophan octyl ester (TOE). This molecule has attracted notable attention as a suitable model for the natural fluorophore tryptophan, in case of membrane proteins. We report here, for the first time, the fluorescence emission behaviour of TOE in reverse micelles of aerosol-OT (AOT) in n-heptane, containing different amounts of water. Relevant studies in representative homogeneous solvent media are also included for comparison. The fluorescence emission parameters (especially emission maximum, relative intensity, and anisotropy) of TOE are found to exhibit significant variation upon changes in the water/surfactant molar ratio (w(0)) of the reverse micelles. Fluorescence decay studies on TOE which we have also performed, indicate biexponential decay kinetics in reverse micelles as well as in homogeneous solvent media. The implications of these findings are examined in relation to the potentialities of TOE as a novel fluorescence probe for membrane proteins present in water restricted environments prevailing at the interfaces of biomembranes (for which reverse micelles serve as ideal model systems). PMID- 11027633 TI - Leptin resistance of adipocytes in obesity: role of suppressors of cytokine signaling. AB - Liver-derived hyperleptinemia induced in normal rats by adenovirus-induced gene transfer causes rapid disappearance of body fat, whereas the endogenous adipocyte derived hyperleptinemia of obesity does not. Here we induce liver-derived hyperleptinemia in rats with adipocyte-derived hyperleptinemia of acquired obesity caused by ventromedial hypothalamus lesioning (VMH rats) or by feeding 60% fat (DIO rats). Liver-derived hyperleptinemia in obese rats caused only a 5 7% loss of body weight, compared to a 13% loss in normoleptinemic lean animals; but in actual grams of weight lost there was no significant difference between obese and lean groups, suggesting that a subset of cells remain leptin-sensitive in obesity. mRNA and protein of a putative leptin-resistance factor, suppressor of cytokine signaling (SOCS)-1 or -3, were both increased in white adipose tissues (WAT) of VMH and DIO rats. Since transgenic overexpression of SOCS-3 in islets reduced the lipopenic effect of leptin by 75%, we conclude that the increased expression of SOCS-1 and -3 in WAT of rats with acquired obesity could have blocked leptin's lipopenic action in the leptin-resistant WAT population. PMID- 11027634 TI - Molecular cloning, genomic structure, chromosomal localization, and alternative splice forms of the platelet collagen receptor glycoprotein VI. AB - Glycoprotein VI (GPVI) is the major collagen receptor underlying platelet activation. We cloned the full-length cDNA for GPVI (GPVI-1) and its two isoforms (GPVI-2 and -3) from phorbol-ester-stimulated CMK cells. The GPVI-1 cDNA was identical in the coding region with the cDNA that has recently been reported to belong to the immunoglobulin superfamily. The GPVI gene consisted of 8 exons spanning over 23 kbp and was mapped on the chromosome 19q13. 4. The promoter of GPVI gene lacked TATA and CAAT boxes and had multiple transcription start sites like other megakaryocytic genes. When COS-7 cells were cotransfected with the GPVI isoforms and Fc receptor gamma chain, Fc receptor gamma chain was associated with GPVI-1 and -2 but did not affect the GPVI expression levels. GPVI-1 and -2 could bind the collagen-related peptide, which exhibits triple-helical and polymeric structure of collagen to activate platelets via GPVI. PMID- 11027635 TI - Monocyte chemotactic protein 1 upregulates IL-1beta expression in human monocytes. AB - Monocyte chemotactic protein-1 (MCP-1) chemoattracts and activates monocytes. The nature of the genes that are transcriptionally activated in the monocytes by MCP 1 is not well understood. To identify such genes, human blood monocytes were incubated with or without MCP-1 for periods of 1, 4, and 12 h and the RNA extracted from these monocytes was subjected to differential display. The differentially expressed transcripts were cloned and sequenced. Differential display showed that interleukin-1beta (IL-1beta) gene expression was upregulated by MCP-1 treatment of monocytes for 4 to 12 h. Quantitative PCR and ELISA assays showed that MCP-1 treatment caused elevation in the levels of IL-1beta transcripts and protein, respectively. Immunoblot analysis showed that most of the protein was pro-IL-1beta. Since IL-1beta is known to induce MCP-1 synthesis, the present demonstration that MCP-1 induces IL-1beta synthesis suggests that the induction of each other would amplify the biological effects of these cytokines during inflammation. PMID- 11027636 TI - Activation of MMP-2 by Clostridium difficile toxin B in bovine smooth muscle cells. AB - Matrix metalloproteinase-2 (MMP-2) plays critical roles in cell migration through the breakdown of the extracellular matrix. Cell movements require dynamic actin reorganization, which is controlled by Rho family GTPases. In order to examine the relation between MMP-2 regulation and actin reorganization, we used several inhibitors of Rho family GTPases. Treatment of smooth muscle cells with Clostridium difficile toxin B known to inactivate Rho family GTPases activated MMP-2. However, neither C3 transferase, a Rho inhibitor, nor Y-27632, a specific inhibitor of Rho-kinase, induced MMP-2 activation. Treatment with C3 transferase and Y-27632 caused morphological changes into the round and stellate shape, respectively, by inhibition of actin stress fiber formation. In addition, toxin B treatment induced expression and processing of MT1-MMP, a major activator of MMP 2. Taken together, we suggest the involvement of Rho family GTPases, although inhibition of neither Rho nor Rho-kinase is sufficient, in the activation of MMP 2 through expression and activation of MT1-MMP. PMID- 11027637 TI - Glycation of aspartate aminotransferase and conformational flexibility. AB - Glycation of proteins alters biological function and changes cellular processes. Our study investigated the conformational changes that accompany glycation using the cardiac aspartate aminotransferase (cAAT). We examined the effects of brief and prolonged exposure of cAAT to glyceraldehyde (Glyc) and ribose 5-phosphate (R5P). When cAAT was briefly incubated (3.5 h) with Glyc (500 microM) or R5P (5 mM) at 37 degrees C, cAAT activity and 1-anilinonaphthalene 8-sulfonate (ANS) binding increased relative to control. After prolonged incubation (64 h) with Glyc (500 microM) or R5P (5 mM) at 37 degrees C, activity and ANS binding decreased relative to control. Furthermore, upon prolonged incubation of cAAT with 500 microM Glyc (14.5 h) or 2 mM R5P (64.25 h) at 37 degrees C, the denaturation curves shifted to the right relative to control. We conclude that upon brief incubation with Glyc and R5P, cAAT exhibited a more open and flexible structure and upon prolonged incubation, a more rigid structure. PMID- 11027638 TI - Anti-PorA antibodies elicited by immunization with peptides conjugated to P64k. AB - To increase the humoral immune response against two cyclic synthetic peptides, derived from variable regions within the outer membrane meningococcal protein PorA (subtypes 19 and 15), we conjugated the peptides to P64k, a novel carrier protein from the same bacterium expressed in Escherichia coli. In addition, one of these peptides was restricted to a linear conformation before it was chemically coupled to the carrier. The conjugates were administered to mice in a three-dose immunization schedule, resulting in a potent anti-peptide immune response, which suggested that chemical conjugation to this carrier provided T cell help. Antisera directed to the three conjugates reacted with Neisseria meningitidis outer membrane PorA upon immunoblot analysis. Moreover, in two out of three conjugates, the anti-peptide sera reacted with native meningococcal outer membrane vesicles in ELISA. PMID- 11027639 TI - Angiogenesis modulators expression in culture cell lines positives for HPV-16 oncoproteins. AB - Altered angiogenesis response is observed in patients with cervical cancer. In this study we examined whether Human Papilloma Virus (HPV) positive epithelial cells are able to produce angiogenic modulators. When added to human umbilical vein endothelial cells (HUVEC) the media conditioned by HPV-16 positive cells was able to induce proliferation, whereas a contrary effect was observed for media derived from non-tumorigenic keratinocytes. The analyses of angiogenesis modulator's mRNA levels result in a decrease of the antiangiogenic factors TSP-1 and 2 in HPV-16 positive cells. In contrast the expression of the pro-angiogenic molecules: bFGF, IL-8, TGF-beta, TNFalpha, and VEGF were higher in these cells as compared to control keratinocytes. Furthermore the pattern of VEGF isoforms observed in the cells positive for the viral genome point to a preferential induction of the VEGF(189) isoform. We therefore conclude that cervical cancer cells expressing HPV-16 genome are able to contribute to the pro-angiogenic response that might support tumor growth and invasion of the surrounding tissues. PMID- 11027640 TI - Enhanced p73 expression during differentiation and complex p73 isoforms in myeloid leukemia. AB - The p53 homologue p73 is expressed in at least six different isoforms (alpha, beta, gamma, delta, epsilon, and zeta), but unlike p53 it has rarely been found mutated in human cancers. However, altered expression of this gene has been reported in cancer cells. In order to understand if p73 is involved in normal and malignant development of myeloid cells, we investigated the expression pattern of the different p73 isoforms in progenitor and mature normal myeloid cells as well as in cells derived from acute and chronic myeloid leukemias. The results show that expression of p73 is markedly enhanced during differentiation of myeloid leukemic cells and that leukemic blasts from patients show an increased expression of the shorter p73 isoforms (gamma, delta, epsilon, zeta). In particular the epsilon isoform is only expressed in leukemic cells and completely absent in mature myeloid cells. Altogether our data suggest that p73 is involved in myeloid differentiation and its altered expression is involved in leukemic degeneration. PMID- 11027641 TI - Suppression of the expression of the CYP2B1/2 gene by retinoic acids. AB - The effects of 5alpha-androsten-3alpha-ol (ASE), and retinoic acids (RAs) and their precursors on the phenobarbital (PB)-mediated induction of CYP2B1 and 2B2 were examined in cultured rat hepatocytes. Two isomers of RA, 9-cis- and all trans-RA, suppressed markedly the effect of PB on CYP2B1/2 expression, while ASE had no suppressive effect. The effect of 9-cis-RA appeared at a lower concentration than the all-trans-isomer, indicating the dominant action of the former isomer. Suppression with 9-cis-retinal was also observed, but all-trans retinol and -retinal were without effect. These results suggest that: (1) ASE, an inverse agonist for the constitutive androstane receptor (CAR), does not play a major role in the suppression of the CYP2B; (2) 9-cis-RA suppresses CYP2B induction by reducing ligand-free retinoid X-receptors (RXR) available for dimerization with the CAR; and (3) enzymes responsible for RA formation play an important role in the mechanism governing CYP2B regulation. PMID- 11027642 TI - Lysophosphatidic acid enhances collagen gel contraction by hepatic stellate cells: association with rho-kinase. AB - We studied the effect of lysophosphatidic acid (LPA) on collagen gel contraction by cultured rat hepatic stellate cells (HSCs) in association with the function of Rho-kinase, one of the target molecules of small GTPase Rho. Binding studies showed a single class-binding site of LPA on HSCs. LPA enhanced the contraction of a collagen lattice seeded with HSCs. LPA increased the number of HSCs with polygonal morphology that contained actin stress fibers, and enhanced the phosphorylation of myosin light chain and the assembly of focal adhesion kinase and RhoA around fibronectin-coated beads seeded on HSCs. The electric cell substrate impedance sensor system showed that LPA enhanced adhesion of HSC to extracellular substrate. All the effects of LPA were suppressed by Y-27632, Rho kinase inhibitor. These data support the notion that LPA is involved in modulating HSC morphology, its attachment to surrounding extracellular matrix and its contraction by a mechanism involving Rho-kinase. PMID- 11027643 TI - Partial characterization of a novel cathepsin L-like protease from Fasciola hepatica. AB - A 30-kDa protease, purified previously from Fasciola hepatica, was sequenced and the first 15 N-terminal residues were found to be 100% homologous to a region in the protein Fcp1c, which was cloned and expressed from F. hepatica. This terminal region was also 53 and 54% identical to two other cathepsin L-like proteases isolated from the same source. The 30-kDa protease demonstrated a specificity different from humancathepsin L when assayed with novel peptidyl enediones of the type Z-Phe-Ala-CH&dbond;CH(2)-CO(2)R (where R = Me/Et/Bu(t)). The ethyl ester peptide was a more efficient inhibitor of the protease than the corresponding methyl ester. This is in contrast to bovine cathepsin B and human cathepsin L where both are more readily inhibited by the methyl, rather than the ethyl ester peptide. These differences in the inhibition of the novel parasite protease may allow it to be exploited as a chemotherapeutic target. PMID- 11027644 TI - Use of domain specific ligands to study urea-induced unfolding of bovine serum albumin. AB - Urea-induced structural transitions in different domains of bovine serum albumin (BSA) were studied fluorometrically using domain specific ligands; chloroform, bilirubin, and diazepam. Urea denaturation of BSA showed a two-step, three-state transition with the accumulation of an intermediate around 4.8-5.2 M urea. During first transition (0-5.0 M urea), a continuous decrease (starting from 1.0 M urea) in diazepam (a ligand for domain III) binding and a late (from 3.0 M urea onward) decrease in chloroform (a ligand primarily for domain I) binding suggested major conformational changes in domain III and partial but significant loss of native conformation in domain I prior to intermediate formation. Absence of any decrease in bilirubin (a ligand for domain II) binding up to 4.5 M urea indicated non involvement of domain II in the unfolding of BSA in this region. However, decrease in bilirubin binding during second transition reflected the unfolding of domain II and its separation from domain I. PMID- 11027645 TI - Complete genomic sequence of a dengue type 2 virus from the French West Indies. AB - Severe forms of dengue fever, dengue haemorrhagic fever, and dengue shock syndrome, were not prominent in the Americas until the epidemic of Cuba in 1981. Since that time, they have spread to other countries in Central and South America, correlating with the spread of dengue type 2 viruses related to Southeast Asian strains. We report here the complete genomic sequence of a dengue type 2 virus isolated during the epidemic in La Martinique in 1998. This constitutes the first complete genetic characterization of a dengue virus strain from French West Indies, and also the first molecular identification in this region of a dengue 2 strain phylogenetically related to the emerging American type 2 dengue viruses. PMID- 11027646 TI - Analysis of cell mechanics in single vinculin-deficient cells using a magnetic tweezer. AB - A magnetic tweezer was constructed to apply controlled tensional forces (10 pN to greater than 1 nN) to transmembrane receptors via bound ligand-coated microbeadswhile optically measuring lateral bead displacements within individual cells. Use of this system with wild-type F9 embryonic carcinoma cells and cells from a vinculin knockout mouse F9 Vin (-/-) revealed much larger differences in the stiffness of the transmembrane integrin linkages to the cytoskeleton than previously reported using related techniques that measured average mechanical properties of large cell populations. The mechanical properties measured varied widely among cells, exhibiting an approximately log-normal distribution. The median lateral bead displacement was 2-fold larger in F9 Vin (-/-) cells compared to wild-type cells whereas the arithmetic mean displacement only increased by 37%. We conclude that vinculin serves a greater mechanical role in cells than previously reported and that this magnetic tweezer device may be useful for probing the molecular basis of cell mechanics within single cells. PMID- 11027647 TI - The study of crystallization of estrogenic 17beta-hydroxysteroid dehydrogenase with DHEA and DHT at elevated temperature. AB - Most crystallization experiments of macromolecules are carried out at a constant temperature. Room temperature (22 degrees C) and 4 degrees C are the most widely used settings in crystallization. In practice, crystal growth at relatively high temperatures has often been avoided for macromolecular crystallization. Human estrogenic 17beta-hydroxysteroid dehydrogenase has been crystallized in complex with dehydroepiandrosterone or dihydrotestosterone. The crystallization experiments were carried out at 27 degrees C. The 17beta-HSD1 crystals were greatly improved at the elevated temperature. The effects of higher temperatures on crystal growth were studied. High temperatures stimulated the nucleation of 17beta-HSD1, increased the rate of crystal growth, and higher occupancy of substrates was obtained in the crystal structure. This method also reduced the formation of twin crystals. Since temperature is the easiest factor to control in the laboratory, crystallization at elevated temperatures provides an efficient method to improve protein crystal growth. The mechanism of the effect of temperature and relative techniques are discussed. PMID- 11027648 TI - Mechanism of ultraviolet B-induced cell cycle arrest in G2/M phase in immortalized skin keratinocytes with defective p53. AB - The ultraviolet B (UVB) portion (280-320 nm) of solar radiation is considered to be a major etiologic factor in human skin cancer and is a known cause of extensive DNA damage. In this study, we observed that UVB exposure of immortalized epidermal keratinocytes (HaCat cells) harboring mutant p53 leads to G(2)/M cell cycle arrest in both asynchronously growing and synchronized cells in a dose dependent manner. Following UVB exposure (200 mJ/cm(2)), we observed a threefold increase in G(2)/M population at 6 h, which increased to sixfold. The observed G(2)/M arrest was associated with an increase in cyclin B level whereas cdc2 protein remained unchanged. However, we observed an accumulation of tyrosine 15 hyperphosphorylated cyclin B-cdc2 complex. In addition, we observed an increase in chk1 kinase and a decrease in cdc25C protein levels. Chk1 phosphorylates cdc25C on serine 216 and inactivates it whereas cdc25C dephosphorylates tyrosine 15 phosphate of cdc2 and activates the cdc2-cyclin B complex. Therefore, the increase in chk1 and the decrease in cdc25C both participate in inhibiting the G2/M transition. Our data identifies two upstream targets leading to inhibition of cyclin B-cdc2 complexes, which explain the inhibition in cyclin B-associated cdc2 kinase following UVB exposure. The inactive phosphorylated cdc2-cyclin B complex remains sequestered in cytoplasm and may migrate to the nucleus following activation. Our data also indicate that UVB exerts unique effects in different types of skin keratinocytes having nonfunctional or mutant p53. PMID- 11027649 TI - Oxygenation of (3Z)-alkenals to 4-hydroxy-(2E)-alkenals in plant extracts: a nonenzymatic process. AB - There is large interest in 4-hydroxy-(2E)-alkenals because of their cytotoxicity in mammals. However, the biosynthetic pathway for these compounds has not been elucidated yet. In plants, 4-hydroxy-(2E)-alkenals were supposed to be derived by the subsequent actions of lipoxygenase and a peroxygenase on (3Z)-alkenals. The presence of 9-hydroxy-12-oxo-(10E)-dodecenoic acid (9-hydroxy-traumatin) in incubations of 12-oxo-(9Z)-dodecenoic acid (traumatin) in the absence of lipoxygenase or peroxygenase, has prompted us to reinvestigate its mode of formation. We show here that in vitro 9-hydroxy-traumatin, 4-hydroxy-(2E)-hexenal and 4-hydroxy-(2E)-nonenal, are formed in a nonenzymatic process. Furthermore, a novel product derived from traumatin was observed and identified as 11-hydroxy-12 oxo-(9Z)-dodecenoic acid. The results obtained here strongly suggest that the 4 hydroxy-(2E)-alkenals, observed in crude extracts of plants, are mainly due to autoxidation of (3Z)-hexenal, (3Z)-nonenal and traumatin. This may have implications for the in vivo existence and previously proposed physiological significance of these products in plants. PMID- 11027650 TI - Increase in cap- and IRES-dependent protein synthesis by overproduction of translation initiation factor eIF4G. AB - The role of eIF4G during the initiation of protein synthesis was studied using mouse mammary carcinoma FM3A cells and FM4G cells that overproduce an N terminally truncated form of eIF4G, which lacks the binding site of poly(A) binding protein. An increase in eIF4G was correlated with an increase in protein synthesis and RNA helicase activity. Translation of mRNAshaving both short and long 5'-untranslated regions (5'-UTR) increased significantly in FM4G cells compared to that in FM3A cells. Both full-length and N-terminally truncated eIF4G transfectants of NIH3T3 cells formed colonies in soft agar and increased the saturation density of cell growth, indicating that both eIF4Gs function similarly. We also found that an internal ribosome entry site (IRES) exists in the 5'-UTR of ornithinedecarboxylase mRNA and that IRES-dependent protein synthesis increased in FM4G cells. Our results indicate that an increase in eIF4G contributes to the formation of active eIF4F similarly to that caused by an increase in eIF4E, as well as to a stimulation of IRES-dependent protein synthesis. PMID- 11027651 TI - Altered gene expression upon BCR cross-linking in Burkitt's lymphoma B cell line. AB - Burkitt's lymphoma cell line, BL2 was stimulated by surface BCR cross-linking and altered gene expression was analyzed by RDA methodology. Consistent with previous reports, we detected up-regulated MDC, IL6R and adhesion molecule LFA1. We also detected gene expression of SIRPalpha, anti-apoptotic A-20, signal regulatory SLP76 and BCAR3, DNA binding proteins EGR2 and DEC1 in addition to some new genes. PMID- 11027652 TI - Arachidonic acid causes cytochrome c release from heart mitochondria. AB - Arachidonic acid interaction with heart mitochondria is known to cause uncoupling as well as inhibition of pyruvate + malate and succinate-supported respiration. Here we present experiments showing that arachidonic acid causes cytochrome c release from Ca(2+)-loaded heart mitochondria. We have also measured mitochondrial matrix swelling and found a fairly good correlation between the two processes, as revealed by the same arachidonic acid concentration dependence and by the same susceptibility toward different free fatty acid species. The effects produced by arachidonic acid are not related to its protonophoric activity since, under the experimental conditions used, saturating concentrations of FCCP did not cause any effect. PMID- 11027653 TI - Exocytosis in the dissociated pancreatic acinar cells of the guinea pig directly visualized by VEC-DIC microscopy. AB - To elucidate the detailed process of exocytosis at the highest possible accuracy, we dissociated the pancreatic acinus of the guinea pig and observed zymogen granules under a video-enhanced contrast differential interference contrast (VEC DIC) microscope. The preparation was thin enough to resolve each zymogen granule with the best clarity. When acinar cells were stimulated with ACh (20 microM), many zymogen granules near the lumen showed an abrupt light intensity change. For a period of 10 s immediately before exocytosis, zymogen granules neither shifted their position nor altered their shape within an accuracy of 38 nm. The time required for individual granules to change the light intensity (the releasing time) ranged from 0.15 to 0.70 s. After each response, the granule maintained its altered contrast for a few seconds until it was retrieved to a planar membrane. No compound exocytosis including granule-granule fusion was observed. We concluded that the exocytosis is not directly initiated by any supramolecular change but by a purely molecular event. PMID- 11027654 TI - An unknown endogenous inhibitor of Na/Ca exchange can enhance the cardiac muscle contractility. AB - The cardiac sarcolemma Na/Ca exchanger is a key system for controlling the intracellular calcium levels during the excitation-contraction coupling. Here, we test the hypothesis that the heart tissue contains a putative endogenous factor having a capacity to modulate the Na/Ca exchanger and muscle contractility. The concentrated cardiac extracts inhibit the Na(i)- or Ca(i)-dependent (45)Ca uptakes in isolated cardiac sarcolemma vesicles as well as the Na(o)-dependent Ca efflux, monitored by extravesicular Ca probe fluo-3. The inhibitory activity has been purified approximately 2000-fold by normal and reversed-phase HPLC procedures. The inhibitory activity is eluted from the Sephadex G-10 in the range of 350-550 Da, suggesting that the inhibitory factor is a low-molecular-weight substance. The mass spectra analysis shows a number of signals within m/z 380 560; however, it is not clear at this moment whether these recordings represent the mass of putative inhibitory factor or irrelevant impurities. The endogenous inhibitory factor of Na/Ca exchange does not resemble the properties (HPLC retention time, mass spectra, amino acid analysis, etc.) of autoinhibitory XIP peptide. The addition of inhibitory factor to muscle strip of guinea pig ventricles induces 2- to 5-fold enhancement of isometric contractions, thereby exhibiting a strong positive inotropic effect. This effect is a dose-dependent phenomenon, which can be reversed by washing the inhibitory factor from the organ bath. Assuming a molecular weight of 350-550 Da, the effective concentrations of putative inhibitor must be <10(-6) M. Therefore, the present findings demonstrate that the mammalian heart contains a low-molecular-weight factor that can inhibit Na/Ca exchange and enhance the cardiac contractility. PMID- 11027655 TI - Tracing the origin of the RACTK1 K(+) channel. AB - Potassium secretion by the kidney is vital for the maintenance of K(+) homeostasis. RACTK1, a putative inwardly rectifying potassium channel cloned from cultured rabbit collecting duct cells, has been proposed to play a role in this process. However, the lack of homology with any other cloned potassium channel and the inability to reproduce the results across different laboratories has brought into question the existence of RACTK1. Recently, it has been suggested that RACTK1 is a contamination from Escherichia coli. In this work we add conclusive evidence supporting the bacterial origin of RACTK1. Using both genomic PCR and RT-PCR we were unable to detect RACTK1 in a number of mammalian species. In addition sequencing of RACTK1 cDNA confirmed a complete homology between RACTK1 and a region of E. coli genomic DNA. Finally, a hypothesis on how RACTK1 could have been generated from a contamination by E. coli genomic DNA is presented. PMID- 11027657 TI - Suppressive effects of DNA vaccines encoding heat shock protein on Helicobacter pylori-induced gastritis in mice. AB - We investigated the effect of DNA vaccines encoding H. pylori-heat shock protein A and B (pcDNA3.1-hspA and -hspB) on inducing immune responses against H. pylori in mice. C57BL/six mice aged 5 weeks were immunized by single injection of 10 microg of pcDNA3.1-hspA and pcDNA3.1-hspB into intracutaneous tissue. Plasmid DNA lacking the inserted hsp were injected as a control. Three months after vaccination, significant specific antibodies against H. pylori were detected by ELISA in the sera of vaccinated mice. Antibody isotypes were predominantly IgG2a (Th1-like) with pcDNA3.1-hspA and mixed IgG1/IgG2a (Th0-like) with pcDNA3.1-hspB. DNA vaccination dramatically suppressed colonies of bacteria in stomach of vaccinated mice (28,400 +/- 21,600/mm(2) for pcDNA3.1-hspA and 6800 +/- 3470/mm(2) for pcDNA3.1-hspB) compared to control mice (128,000 +/- 42,200/mm(2)). Histological analysis of the gastric mucosa demonstrated that the degree of gastritis was significantly lower in the vaccinated mice than in control mice. These results demonstrated that DNA vaccines encoding H. pylori-Hsp induce significant immune response against H. pylori to decrease gastric mucosal inflammation, indicating that a DNA vaccine can be a new approach against H. pylori in humans. PMID- 11027658 TI - Localization of aquaporin-7 in rat and mouse kidney using RT-PCR, immunoblotting, and immunocytochemistry. AB - To establish the segmental, cellular, and subcellular localization of AQP7 in rat and mouse kidney, we used RT-PCR, immunocytochemical, and immunoblotting approaches. RT-PCR of rat and mouse kidney zones revealed AQP7 mRNA in cortex and outer stripe of the outer medulla. RT-PCR on microdissected nephron segments revealed AQP7 mRNA in proximal convoluted and straight tubules. Immunoblotting using peptide-derived rabbit antibodies to either rat or mouse AQP7 revealed a 28 kDa band in kidney and testes from rat and mouse, respectively. Immunocytochemistry revealed strong AQP7 labeling of segment 3 proximal tubules and weaker labeling of proximal convoluted tubules in both rat and mouse kidneys. The labeling was almost exclusively confined to the brush border with no basolateral labeling. No labeling was observed of thin descending limbs or collecting duct. Immunolabeling controls were negative. The presence of AQP7 in the proximal tubule brush border indicates a role of AQP7 in proximal tubule water reabsorption. PMID- 11027656 TI - Cell density-dependent regulation of hepatic development by a gp130-independent pathway. AB - We previously demonstrated that oncostatin M (OSM) promotes hepatic development in concert with glucocorticoid. The livers from mice deficient for gp130, a signaling subunit of the OSM receptor, displayed reduced expression of hepatic differentiation marker and defective glycogenic function. However, these phenotypes were not completely abolished in gp130(-/-) mice, suggesting that there is an alternative pathway regulating hepatic development in vivo. To test this possibility, we cultured gp130(-/-) fetal hepatic cells and investigated a signal that induces hepatic differentiation. When hepatocytes were forced to interact with each other by inoculating cells at high densities, hepatic differentiation was induced even in the absence of gp130. Moreover, cells stimulated with OSM and/or cultured at a high density possess many other metabolic functions. These observations suggest that fetal hepatic cells acquire multiple characteristics of differentiated hepatocytes in response to the signals generated by cell-cell contacts as well as by OSM. PMID- 11027659 TI - Analysis of the role of conserved cysteine residues in the bcl-2 oncoprotein. AB - The Bcl-2 oncoprotein is an integral membrane protein localized primarily to the outer membrane of the mitochondria. The precise molecular mechanism responsible for the antiapoptotic action of Bcl-2 remains unknown. Two cysteine residues are found in Bcl-2 and these residues are well-conserved across species. The first cysteine (cys(155)) is located in the alpha5 domain, a region important for the ion channel properties of Bcl-2, while the second cysteine (cys(226)) is located in the carboxyl-terminal membrane anchor domain. In this study, we found that replacement of both cysteines with serine residues generated a mutant protein that retained the ability to homodimerize and heterodimerize with proapoptotic Bax protein in vitro. In whole cells, the mutant protein efficiently heterodimerized with Bax, but exhibited impaired homodimerizationrelative to wild type Bcl-2. The mutant protein was also less efficient than wild-type Bcl-2 at suppressing caspase activation, DNA fragmentation, and loss of viability during IL-3 withdrawal-induced apoptosis. Together, the data indicate that the cysteine residues in Bcl-2 contribute, but are not absolutely essential, to the ability of Bcl-2 to homodimerize, heterodimerize with Bax, and suppress apoptosis. PMID- 11027660 TI - Arsenic trioxide inhibits neuroblastoma growth in vivo and promotes apoptotic cell death in vitro. AB - Recent clinical studies have shown that inorganic arsenic trioxide (As(2)O(3)) at low concentrations induces complete remission with minimal toxicity in patients with refractory acute promyelocytic leukemia (APL). Preclinical studies suggest that As(2)O(3) induces apoptosis and possibly differentiation in APL cells. Like APL cells, neuroblastoma (NB) cells are thought to be arrested at an early stage of differentiation, and cells of highly malignant tumors fail to undergo spontaneous maturation. Both APL and NB cells can respond with differentiation to retinoic acid (RA) treatment in vitro and probably also in vivo. For that reason we investigated the effect of As(2)O(3) alone and in combination with RA on NB cell lines. In vitro, the number of viable NB cells was reduced at As(2)O(3) concentrations around 1 microM after 72 h exposure. The IC50 in six different cell lines treated for 3 days was in the 1.5 to 5 microM concentration interval, the most sensitive being SK-N-BE(2) cells derived from a chemotherapy resistant tumor. The combined treatment with RA (1 and 3 microM) showed no consistent additional effect with regard to induced cell death. The effect of As(2)O(3) on NB cell number involved As(2)O(3)-induced apoptotic pathways (decreased expression of Bcl-2 and stimulation of caspase-3 activity) with no clear evidence of induced differentiation. The in vivo effect of As(2)O(3) on NB growth was also investigated in nude mice bearing tumors of xenografted NB cells. Although tumor growth was reduced by As(2)O(3) treatment, complete remission was not achieved at the concentrations tested. We suggest that As(2)O(3), in combination with existing treatment modalities, might be a treatment approach for high risk NB patients. PMID- 11027661 TI - Kringle 1 of human hepatocyte growth factor inhibits bovine aortic endothelial cell proliferation stimulated by basic fibroblast growth factor and causes cell apoptosis. AB - Hepatocyte growth factor (HGF), also known as scatter factor, is a mesenchymal or stromal-derived mediator with angiogenic activity. There are four kringle domains in its amino terminus. They display considerable sequence similarity with those of angiostatin, an angiogenesis inhibitor. We now describe that the recombinant kringle1 of HGF (HGFK1) inhibits bovine aortic endothelial (BAE) cell proliferation stimulated by basic fibroblast growth factor in a dose-dependent manner, with an ED(50) of approximately 0.7 microg/ml, while ED(50) of angiostatin is 3 microg/ml. Treatment of BAE cell with HGFK1 caused cell apoptosis. This report thus constitutes the first demonstration that kringle1 of HGF is a selective inhibitor for BAE cell proliferation stimulated by bFGF. PMID- 11027662 TI - Central effects of neuromedin U in the regulation of energy homeostasis. AB - Neuromedin U (NMU) is a brain-gut peptide whose peripheral activities are well understood but whose central actions have yet to be clarified. The recent identification of two NMU receptors in rat brain has provided a springboard for further investigation into its role in the central nervous system. Intracerebroventricular administration of NMU to free-feeding rats decreased food intake and body weight. Conversely, NMU increased gross locomotor activity, body temperature, and heat production. NMU, a potent endogenous anorectic peptide, serves as a catabolic signaling molecule in the brain. Further investigation of the biochemical and physiological functions of NMU will help our better understanding of the mechanisms of energy homeostasis. PMID- 11027663 TI - Flt3 ligand induces tyrosine phosphorylation of gab1 and gab2 and their association with shp-2, grb2, and PI3 kinase. AB - The receptor tyrosine kinase Flt3 has been shown to play an important role in proliferation, differentiation, and survival of hematopoietic stem and progenitor cells. Although some postreceptor signaling events of Flt3 have been characterized, the involvement of Gab family proteins in Flt3 signaling is not known. In this study, we show that both Gab1 and Gab2 are rapidly tyrosine phosphorylated after Flt3 ligand stimulation of Flt3 ligand-responsive cells. They interact with tyrosine-phosphorylated Shp-2, p85, Grb2, and Shc. The results suggest that Gab proteins are engaged in Flt3 signaling to mediate downstream activation of Shp-2 and PI3 kinase pathways and possibly the Ras/Raf/MAPK pathway. PMID- 11027664 TI - Genomic organization and expression of the mouse equilibrative, nitrobenzylthioinosine-sensitive nucleoside transporter 1 (ENT1) gene. AB - We have cloned and characterized the genomic structure of the mouse gene for the NBMPR-sensitive equilibrative nucleoside transporter (mENT1), which is located on chromosome 17C. About 12-kb of genomic DNA was sequenced including the promoter region, 12 exons, 11 introns, and the 3'-untranslated region. All exon-intron junction sequences conform to the GT/AG rule. Primer extension analysis demonstrated a transcription initiation site located 252 bp upstream of the translation start site. Analysis of the 2.5-kb 5'-flanking sequence shows putative binding sites for several transcription factors, including GATA-1, IRF 2, Pit-1, myogenin, CREB, Sp-1, Ap-2, MAZ, and GR. We demonstrated that mouse ENT1 mRNA was highly expressed in heart, spleen, lung, liver, and testis. Lower levels of expression were detected in brain and kidney. Functional analysis of the 5'-flanking region showed that the nucleotide sequence from -652 to -111 contains cis-regulatory elements that promote gene expression. We found two Sp-1 binding sites (-296/-303, -307/-313) and two MAZ binding sites (-353/-359, -522/ 528) in this region. Luciferase assay results suggest that MAZ and Sp-1 transcription factors are important positive regulators of transcription for the ENT1 gene in NG108-15 cells. PMID- 11027665 TI - Molecular cloning and functional characterization of a novel nuclear receptor similar to an embryonic benzoate receptor BXR. AB - Benzoate X receptor (BXR) is a member of the nuclear hormone receptor superfamily and is activated by alkyl esters of amino benzoic acids and expressed during early development stages. We report here a second BXR, which was cloned by screening Xenopus laevis embryonic cDNA libraries. This new BXR, termed BXRbeta, exhibits 97% identity in the DNA-binding domain and 79% identity in the ligand binding domain in amino acid sequence to previously reported Xenopus BXR. The BXRbeta strongly binds as a heterodimer with retinoid X receptor (RXR) to direct repeats of an AGGTCA motif spaced 4 or 5 base pairs apart and activates transcription by addition of methyprednisolone and dexamethasone as well as amino and hydroxybenzoates. PMID- 11027666 TI - Requirement for the COOH-terminal pro-sequence in the translocation of aqualysin I across the cytoplasmic membrane in Escherichia coli. AB - Aqualysin I from Thermus aquaticus YT-1 is an extracellular subtilisin-type serine protease. The protease is synthesized as a distinct precursor composed of four functional domains: an N-terminal signal sequence, an N-terminal pro sequence, a protease domain, and a C-terminal pro-sequence. The N-terminal pro sequence is essential for the production of active aqualysin I while the C terminal pro-sequence is required for extracellular secretion of aqualysin I. In an E. coli expression system, the function of C-terminal pro-sequence in the translocation of aqualysin I across the cytoplasmic membrane was investigated. More than 60-70% of the total activity was detected in the cytoplasmic fraction in the deletion mutations of the C-terminal pro-sequence while less than 30% was found in this fraction in wild type. In addition, in vitro processing of aqualysin I precursors with these mutations to a mature form promptly occurred and the folding into active aqualysin I was rapid. These results suggest that the C-terminal pro-sequence, probably in conjunction with the signal sequence, facilitates the translocation of the precursor across the cytoplasmic membrane by preventing the precursor from taking on an active conformation. PMID- 11027667 TI - Calcium increases apolipoprotein B mRNA editing. AB - ApoB-100 and apoB-48 are major components of chylomicrons, very low-density lipoprotein (VLDL) and low-density lipoprotein (LDL). The two proteins are generated from a single apoB mRNA by apoB mRNA editing which induces an in-frame stop codon in apoB mRNA. Apolipoprotein B (apoB) mRNA editing is an important determinant of the proportion of full-length (apoB-100) and truncated (apoB-48) proteins in total apoB metabolism. Calcium is involved in the regulation of secretion and synthesis of VLDL and apoB. In this paper, we demonstrate for the first time that the amount of edited apoB mRNA in the cultured cells Caco-2 and McA7777 is markedly increased by calcium. Increasing extracellular calcium concentration, calcium ionophore (A23187 and ionomycin) treatment, and depleting calcium stores and raising cytoplasmic calcium concentration by thapsigargin increase apoB mRNA editing up to threefold in a dose dependent manner. Calcium has no direct stimulative effect on apoB mRNA editing in an in vitro editing system. The editing increase by extracellular calcium is not related to alterations of APOBEC-1 mRNA expression. These data suggest that calcium is not only involved in the regulation of apolipoprotein metabolism but also apoB mRNA editing. PMID- 11027668 TI - Heat shock protein 60 is a high-affinity high-density lipoprotein binding protein. AB - A new 55-kDa HDL/apolipoprotein binding protein was demonstrated in plasma membrane preparations of the human cell lines and primary cultured hepatocytes. Analysis of specific binding by ligand immunoblots of HDL, apoA-I, and apoA-II to a partially purified 55-kDa PA-I plasma membrane preparation demonstrated a K(d,HDL) = 50 nM (10 microg/ml), K(d,apoA-II) = 20 nM (0.4 microg/ml), and K(d, apoA-I) = 330 nM (10 microg/ml). Following preparative SDS-PAGE electrophoresis of a plasma membrane preparation isolated from human PA-I cells, fractions with apoA-II binding activity were collected, concentrated, and subjected to two dimensional electrophoresis. Internal microprotein sequencing of the 55-kDa protein band revealed the binding protein as being heat shock protein 60 (hsp60). The hsp60 monoclonal antibody LK-1 blocked apoA-II binding to the 55-kDa HBP preparation. In summary, these results provide a potential mechanism to explain the known association between immunity developed against hsp60 and the development of atherosclerosis. PMID- 11027669 TI - Expression profiling of human sulfotransferase and sulfatase gene superfamilies in epithelial tissues and cultured cells. AB - The bioavailability of drugs administered topically or orally depends on their metabolism by epithelial enzymes such as the cytosolic sulfotransferases (SULT). Reverse transcriptase-polymerase chain reaction (RT-PCR) methods were established to detect expression of 8 SULT genes and 4 arylsulfatase (ARS) genes in human tissues of epithelial origin and in cultures of normal and transformed (cancer) cells. The results indicate: (i) SULT 1A1, 1A3, ARSC, and ARSD genes are ubiquitously expressed; (ii) expression is frequently similar between cell lines and corresponding tissues; (iii) SULT gene expression in normal cultured cells is generally comparable to the expression in associated transformed (cancer) cell lines; (iv) SULT 1A1 promoter usage is mainly tissue specific; however, both promoters are frequently used in SULT 1A3 expression; and (v) the expression profile of SULT 1A1, 1A3, 1E1, and 2B1a/b suggests that one or more of these isoforms may be involved in the cutaneous sulfoconjugation of minoxidil and cholesterol. PMID- 11027670 TI - Detection of cellular receptors specific for the hepatitis B virus preS surface protein on cell lines of extrahepatic origin. AB - Hepatitis B virus infection is primarily mediated by the interaction of the preS region of the viral envelope protein with its still unknown cellular receptor. Using recombinantly expressed preS proteins, the distribution of preS-binding receptors on cell lines from extrahepatic origins was determined by immunofluorescence and flow cytometry. In contrast to human liver cell lines, most cell lines from extrahepatic origins did not bind preS proteins. Nevertheless, exceptions were found in the bone marrow-derived cell line, KG-1, and the osteogenic sarcoma cell line SaOS-2, as well as in the previously reported EBV-transformed B-cell line, Wa. To determine the biochemical nature of these receptors, Wa-cells were cell surface biotinylated and the preS-binding receptors were isolated by immunoprecipitation. A specific band with a molecular weight of approximately 30 kDa was identified in a SDS-polyacrylamide gel, which further characterization is expected to provide clues regarding the infection mechanism of HBV in hepatic- and extra-hepatic cells. PMID- 11027671 TI - Intracellular FGF-2 promotes differentiation in T-47D breast cancer cells. AB - To test the implicated role of basic fibroblast growth factor (bFGF; FGF-2) in promoting differentiation in breast cancer, we enforced the expression of FGF-2 in T-47D breast cancer cells. Expression of FGF-2 conferred an overall less malignant phenotype to T-47D cells as revealed by their reduced proliferative response, impaired capacity for anchorage-independent growth, and invasion through Matrigel. To understand one candidate mechanism for the intracellular FGF 2-mediated anti-invasive effect, we examined the effect of FGF-2 on T-47D cell motility. Addition of recombinant FGF-2 to the growth medium markedly enhanced cell motility while constitutive expression of intracellular FGF-2 significantly inhibited the migratory potential of T-47D cells in a dominant manner. FGF-2 expressing T-47D cells also formed relatively defined branching structures in Matrigel matrices, a characteristic phenotype of differentiation in breast cancer cells. These data suggest a potential role for FGF-2 in promoting functional differentiation of breast epithelial cells. PMID- 11027672 TI - The presenilin 1 C92S mutation increases abeta 42 production. AB - Although wild-type human presenilin 1 (PS1) rescues the C. elegans egg-laying (egl) phenotype that is caused by a loss of function mutation in the C. elegans presenilin homologue sel12, most familial Alzheimer's disease (FAD)-linked PS1 mutants only partially rescue this phenotype. To investigate the effects of the loss of function sel12 mutation on Abeta production in mammalian cells, we analyzed Abeta production in transfected H4 neuroglioma cells expressing the PS1 homologue of the sel12 C60S mutant, PS1 C92S. This analysis revealed that PS1 C92S increased Abeta42 levels in a similar fashion to other pathogenic Alzheimer's disease (AD) PS1 mutations. Significantly, the PS1 C92S mutation has recently been identified as the pathogenic mutation in an Italian family with FAD. Thus, placing a mutation that results in loss of function in C. elegans into a context whereby its effect on mammalian cells can be evaluated suggests that all FAD-linked PS1 mutants result in increased Abeta42 production through a partial loss of function mechanism. PMID- 11027673 TI - Neuropeptide Y receptor gene y6: multiple deaths or resurrections? AB - The neuropeptide Y family of G-protein-coupled receptors consists of five cloned members in mammals. Four genes give rise to functional receptors in all mammals investigated. The y6 gene is a pseudogene in human and pig and is absent in rat, but generates a functional receptor in rabbit and mouse and probably in the collared peccary (Pecari tajacu), a distant relative of the pig family. We report here that the guinea pig y6 gene has a highly distorted nucleotide sequence with multiple frame-shift mutations. One evolutionary scenario may suggest that y6 was inactivated before the divergence of the mammalian orders and subsequently resurrected in some lineages. However, the pseudogene mutations seem to be distinct in human, pig, and guinea pig, arguing for separate inactivation events. In either case, the y6 gene has a quite unusual evolutionary history with multiple independent deaths or resurrections. PMID- 11027674 TI - Progesterone receptor activation mediates LH-induced type-I pituitary adenylate cyclase activating polypeptide receptor (PAC(1)) gene expression in rat granulosa cells. AB - We have previously reported that the pituitary adenylate cyclase activating polypeptide (PACAP) gene is regulated in ovarian granulosa cells by the autocrine and/or paracrine interaction between progesterone and its nuclear receptor progesterone receptor (PR). To initiate studies on the functional significance of the progesterone-induced PACAP production in luteinizing granulosa cells, we sought to determine the expression and hormonal regulation of PACAP receptors in the rat ovary. The relative mRNA levels of three known PACAP receptor subtypes (PAC(1), VPAC(1), and VPAC(2)) were determined in ovaries of immature rats treated with gonadotropins, by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) assays. Results show that all PAC(1), VPAC(1), and VPAC(2) transcripts are expressed at a detectable level in immature rat ovaries. Importantly, the ovarian level of PAC(1), but not VPAC(1) or VPAC(2), mRNA notably changes during gonadotropin challenges. Ovarian PAC(1) mRNA expression decreases during the pregnant mare's serum gonadotropin (PMSG)-induced follicular phase but substantially increases during the human chorionic gonadotropin (hCG) induced periovulatory period. Because the hCG-induced increase in ovarian PAC(1) mRNA expression is attributable to the hormone-induced PAC(1) mRNA expression in granulosa cells of the preovulatory follicles, we next examined whether hCG regulates PAC(1) mRNA expression by directly acting on granulosa cells. When granulosa cells isolated from PMSG (40 h)-primed immature rats were challenged with hCG (or forskolin), PAC(1), but not VPAC(1) or VPAC(2), mRNA expression significantly increased within 6 h. Because the LH-induced PAC(1) mRNA expression (6 h) proceeds PR activation (3 h) in granulosa cells as the LH-induced PACAP mRNA expression (6 h) does, we further determined the cause-effect relationship among LH, PR activation and PAC(1) receptor gene expression, by examining the effect of PR antagonist, ZK98299, on the ability of LH to increase PAC(1) mRNA levels in luteinizing granulosa cells. Results show that ZK98299 inhibited the stimulatory effect of hCG (or forskolin) on PAC(1) mRNA expression, at the level of all known splice variants of PAC(1) mRNA in granulosa cells. In summary, our results demonstrating that PR activation is critical for the LH-induced PAC(1) gene expression in luteinizing granulosa cells suggest that PR activation regulates the finely tuned expression of the PACAP/PACAP receptor genes in luteinizing granulosa cells and thus dictates the timing of the autocrine and/or paracrine function of PACAP in preovulatory follicles. PMID- 11027675 TI - The lipids C2- and C16-ceramide form large stable channels. Implications for apoptosis. AB - We report that physiological concentrations of both short- and long-chain ceramides, despite being lipids, form large stable pores in membranes. Some of these pores should be large enough to allow cytochrome c to permeate. Dihydroceramide differs from ceramide by the reduction of one double bond, and yet both its apoptogenic and channel-forming activities are greatly reduced. A structural model provides insight into how ceramides might form pores. According to a mathematical model, both the individual conductance of the channels and the overall membrane conductance are directly related to the overall concentration of ceramide in the membrane. Slight changes in concentration have dramatic effects on the size of the channels formed, providing an easy way for rapidly altering membrane permeability by changing the activity of local synthetic and catabolic enzymes. A possible role for these channels in apoptosis is discussed. PMID- 11027676 TI - Comparison of the interactions of transferrin receptor and transferrin receptor 2 with transferrin and the hereditary hemochromatosis protein HFE. AB - The transferrin receptor (TfR) interacts with two proteins important for iron metabolism, transferrin (Tf) and HFE, the protein mutated in hereditary hemochromatosis. A second receptor for Tf, TfR2, was recently identified and found to be functional for iron uptake in transfected cells (Kawabata, H., Germain, R. S., Vuong, P. T., Nakamaki, T., Said, J. W., and Koeffler, H. P. (2000) J. Biol. Chem. 275, 16618-16625). TfR2 has a pattern of expression and regulation that is distinct from TfR, and mutations in TfR2 have been recognized as the cause of a non-HFE linked form of hemochromatosis (Camaschella, C., Roetto, A., Cali, A., De Gobbi, M., Garozzo, G., Carella, M., Majorano, N., Totaro, A., and Gasparini, P. (2000) Nat. Genet. 25, 14-15). To investigate the relationship between TfR, TfR2, Tf, and HFE, we performed a series of binding experiments using soluble forms of these proteins. We find no detectable binding between TfR2 and HFE by co-immunoprecipitation or using a surface plasmon resonance-based assay. The affinity of TfR2 for iron-loaded Tf was determined to be 27 nm, 25-fold lower than the affinity of TfR for Tf. These results imply that HFE regulates Tf-mediated iron uptake only from the classical TfR and that TfR2 does not compete for HFE binding in cells expressing both forms of TfR. PMID- 11027677 TI - Sp3 is a transcriptional repressor of transforming growth factor-beta receptors. AB - MCF-7E breast cancer cells express transforming growth factor-beta (TGF-beta) receptors RI and RII in comparison to MCF-7L cells. We present data showing that Sp3 acts as a transcriptional repressor of RI and RII in MCF-7L cells and GEO colon cancer cells. MCF-7L and GEO cells express high levels of Sp3 protein. Gel shift analysis indicated enhanced binding of Sp3 from MCF-7L cells to a consensus Sp1 oligonucleotide. Southwestern data indicated increased binding of Sp3 to RI and RII promoters in MCF-7L cells, suggesting a correlation between Sp3 binding and reduced expression of TGF-beta receptors in MCF-7L cells. Cotransfection of CMV-Sp3 cDNA with RI and RII promoter-luciferase reporter constructs decreased RI and RII promoter activities by 70% in MCF-7E and GEO cells. Southwestern analysis detected the binding of transiently expressed Sp3 to RI and RII promoters in MCF 7E cells. Significantly, ectopic Sp3 expression led to repression of RI and RII transcripts in MCF-7E cells. This report demonstrates that inappropriate overexpression of Sp3 is a mechanism that contributes to repression of TGF-beta receptors. PMID- 11027679 TI - Single-stranded DNA-binding complex involved in transcriptional regulation of mouse mu-opioid receptor gene. AB - Previously, we reported the presence of dual (distal and proximal) promoters in mouse mu-opioid receptor (mor) gene, with mor transcription in mouse brain predominantly initiated by the proximal promoter. Sp factors, bound to double stranded (ds) cis-regulatory elements, are critical for proximal promoter activity. Here, we further report that a single-stranded (ss) cis-regulatory element and trans-acting protein factor are also important for proximal promoter activity. A 26-bp mor polypyrimidine/polypurine region (PPy/u) can adopt ss DNA conformation, as demonstrated by S1 nuclease sensitivity. Using electrophoretic mobility shift analysis with nuclear extracts from mor-expressing SH-SY5Y cells, we demonstrate that the sense strand of PPy/u interacts with a major nuclear protein, termed mor polypyrimidine-binding protein (mPy), which is not related to Sp factors. Southwestern blot analysis indicated that mPy protein is approximately 25 kDa in size. Functional analysis suggests that mPy protein can trans-activate mor promoter as well as a heterologous promoter. Moreover, combinatorial activation of ss (mPy) and ds (Sps) DNA binding factors, interacting with an overlapping DNA (PPy/u) region, is necessary for proximal promoter activation. Thus our results suggest that transcription of mouse mor gene is regulated by an interplay of ss and ds DNA binding factors. PMID- 11027678 TI - Mode of action of an antiviral peptide from HIV-1. Inhibition at a post-lipid mixing stage. AB - DP178, a synthetic peptide corresponding to a segment of the transmembrane envelope glycoprotein (gp41) of human immunodeficiency virus, type 1 (HIV-1), is a potent inhibitor of viral infection and virus-mediated cell-cell fusion. Nevertheless, DP178 does not contain gp41 coiled-coil cavity binding residues postulated to be essential for inhibiting HIV-1 entry. We find that DP178 inhibits phospholipid redistribution mediated by the HIV-1 envelope glycoprotein at a concentration 8 times greater than that of solute redistribution (the IC(50) values are 43 and 335 nm, respectively). In contrast, C34, a synthetic peptide which overlaps with DP178 but contains the cavity binding residues, did not show this phenomenon (11 and 25 nm, respectively). The ability of DP178 to inhibit membrane fusion at a post-lipid mixing stage correlates with its ability to bind and oligomerize on the surface of membranes. Furthermore, our results are consistent with a model in which DP178 inhibits the formation of gp41 viral hairpin structure at low affinity, whereas C34 inhibits its formation at high affinity: the failure to form the viral hairpin prevents both lipid and solute from redistributing between cells. However, our data also suggest an additional membrane-bound inhibitory site for DP178 in the ectodomain of gp41 within a region immediately adjacent to the membrane-spanning domain. By binding to this higher affinity site, DP178 inhibits the recruitment of several gp41-membrane complexes, thus inhibiting fusion pore formation. PMID- 11027680 TI - A novel site on the Galpha -protein that recognizes heptahelical receptors. AB - Specific domains of the G-protein alpha subunit have been shown to control coupling to heptahelical receptors. The extreme N and C termini and a region between alpha4 and alpha5 helices of the G-protein alpha subunit are known to determine selective interaction with the receptors. The metabotropic glutamate receptor 2 activated both mouse Galpha(15) and its human homologue Galpha(16), whereas metabotropic glutamate receptor 8 activated Galpha(15) only. The extreme C-terminal 20 amino acid residues are identical between the Galpha(15) and Galpha(16) and are therefore unlikely to be involved in coupling selectivity. Our data reveal two regions on Galpha(16) that inhibit its coupling to metabotropic glutamate receptor 8. On a three-dimensional model, both regions are found in a close proximity to the extreme C terminus of Galpha(16). One module comprises alpha4 helix, alpha4-beta6 loop (L9 Loop), beta6 sheet, and alpha5 helix. The other, not described previously, is located within the loop that links the N terminal alpha helix to the beta1 strand of the Ras-like domain of the alpha subunit. Coupling of Galpha(16) protein to the metabotropic glutamate receptor 8 is partially modulated by each module alone, whereas both modules are needed to eliminate the coupling fully. PMID- 11027681 TI - Purification, cloning, and characterization of a profibrinolytic plasminogen binding protein, TIP49a. AB - The plasminogen receptors responsible for enhancing cell surface-dependent plasminogen activation expose COOH-terminal lysines on the cell surface and are sensitive to proteolysis by carboxypeptidase B (CpB). We treated U937 cells with CpB, then subjected membrane fractions to two-dimensional gel electrophoresis followed by ligand blotting with (125)I-plasminogen. A 54-kDa protein lost the ability to bind (125)I-plasminogen after treatment of intact cells and was purified by two-dimensional gel electrophoresis and then sequenced by mass spectrometry. Two separate amino acid sequences were obtained and were identical to sequences contained within human and rat TIP49a. The cDNA for the 54-kDa protein matched the human TIP49a sequence, and encoded a COOH-terminal lysine, consistent with susceptibility to CpB. Antibodies against rat TIP49a recognized the plasminogen-binding protein on two-dimensional Western blots of U937 cell membranes. Human (125)I-Glu-plasminogen bound specifically to TIP49a protein, and binding was inhibited by epsilon-aminocaproic acid. A single class of binding sites was detected, and a K(d) of 0.57 +/- 0.14 microm was determined. TIP49a enhanced plasminogen activation 8-fold compared with the BSA control, and this was equivalent to the enhancement mediated by plasmin-treated fibrinogen. These results suggest that TIP49a is a previously unrecognized plasminogen-binding protein on the U937 cell surface. PMID- 11027682 TI - Structural basis for lipoxygenase specificity. Conversion of the human leukocyte 5-lipoxygenase to a 15-lipoxygenating enzyme species by site-directed mutagenesis. AB - Mammalian lipoxygenases constitute a heterogeneous family of lipid-peroxidizing enzymes, and the various isoforms are categorized with respect to their positional specificity of arachidonic acid oxygenation into 5-, 8-, 12-, and 15 lipoxygenases. Structural modeling suggested that the substrate binding pocket of the human 5-lipoxygenase is 20% bigger than that of the reticulocyte-type 15 lipoxygenase; thus, reduction of the active-site volume was suggested to convert a 5-lipoxygenase to a 15-lipoxygenating enzyme species. To test this "space based" hypothesis of the positional specificity, the volume of the 5-lipoxygenase substrate binding pocket was reduced by introducing space-filling amino acids at critical positions, which have previously been identified as sequence determinants for the positional specificity of other lipoxygenase isoforms. We found that single point mutants of the recombinant human 5-lipoxygenase exhibited a similar specificity as the wild-type enzyme but double, triple, and quadruple mutations led to a gradual alteration of the positional specificity from 5S- via 8S- toward 15S-lipoxygenation. The quadruple mutant F359W/A424I/N425M/A603I exhibited a major 15S-lipoxygenase activity (85-95%), with (8S,5Z,9E,11Z,14Z)-8 hydroperoxyeicosa-5,9 ,11, 14-tetraenoic acid being a minor side product. These data indicate the principle possibility of interconverting 5- and 15 lipoxygenases by site-directed mutagenesis and appear to support the space-based hypothesis of positional specificity. PMID- 11027683 TI - Functional analysis in type Ia group B Streptococcus of a cluster of genes involved in extracellular polysaccharide production by diverse species of streptococci. AB - Several species of streptococci produce extracellular polysaccharides in the form of secreted exopolysaccharides or cell-associated capsules. Although the biological properties and repeating unit structures of these polysaccharides are diverse, sequence analysis of the genes required for their production has revealed a surprising degree of conservation among five genes found in the capsule gene cluster of each of several polysaccharide-producing streptococci. To determine the function of these conserved genes, we characterized a series of isogenic mutants derived from a wild-type strain of type Ia group B Streptococcus by selectively inactivating each gene. Inactivation of cpsIaE resulted in an acapsular phenotype, consistent with previous work that identified the cpsIaE product as the glycosyltransferase that initiates synthesis of the polysaccharide repeating unit. Mutants in cpsIaA, cpsIaB, cpsIaC, or cpsIaD produced type Ia capsular polysaccharide, but in reduced amounts compared with the wild type. Analysis of the mutant polysaccharides and of capsule gene transcription in the mutant strains provided evidence that cpsIaA encodes a transcriptional activator that regulates expression of the capsule gene operon. Mutants in cpsIaC or cpsIaD produced polysaccharide of reduced molecular size but with an identical repeating unit structure as the wild-type strain. We conclude that CpsA to -D are not required for polysaccharide repeating unit biosynthesis but rather that they direct the coordinated polymerization and export of high molecular weight polysaccharide. PMID- 11027684 TI - Thyroid hormone stimulates acetyl-coA carboxylase-alpha transcription in hepatocytes by modulating the composition of nuclear receptor complexes bound to a thyroid hormone response element. AB - Triiodothyronine (T3) stimulates a 7-fold increase in transcription of the acetyl CoA carboxylase-alpha (ACCalpha) gene in chick embryo hepatocytes. Here, we characterized an ACCalpha T3 response element (ACCalpha-T3RE) with unique functional and protein binding properties. ACCalpha-T3RE activated transcription both in the absence and presence of T3, with a greater activation observed in the presence of T3. In nuclear extracts from hepatocytes incubated in the absence of T3, ACCalpha-T3RE bound protein complexes (complexes 1 and 2) containing the liver X receptor (LXR) and the retinoid X receptor (RXR). In nuclear extracts from hepatocytes incubated in the presence of T3 for 24 h, ACCalpha-T3RE bound a different set of complexes. One complex contained LXR and RXR (complex 3) and another contained the nuclear T3 receptor (TR) and RXR (complex 4). Mutations of ACCalpha-T3RE that inhibited the binding of complexes 1 and 2 decreased transcriptional activation in the absence of T3, and mutations of ACCalpha-T3RE that inhibited the binding of complexes 3 and 4 decreased transcriptional activation in the presence of T3. The stimulation of ACCalpha transcription caused by T3 was closely associated with changes in the binding of complexes 1-4 to ACCalpha-T3RE. These data suggest that T3 regulates ACCalpha transcription by a novel mechanism involving changes in the composition of nuclear receptor complexes bound to ACCalpha-T3RE. We propose that complexes containing LXR/RXR ensure a basal level of ACCalpha expression for the synthesis of structural lipids in cell membranes and that complexes containing LXR/RXR and TR/RXR mediate the stimulation of ACCalpha expression caused by T3. PMID- 11027685 TI - Phage display epitope mapping of human neutrophil flavocytochrome b558. Identification of two juxtaposed extracellular domains. AB - Despite extensive experimental and clinical evidence demonstrating the critical role of flavocytochrome b558 (Cyt b) in the NADPH-dependent oxidase, there is a paucity of direct structural data defining its topology in the phagocyte membrane. Unlike other Cyt b-specific monoclonal antibodies, 7D5 binds exclusively to an extracellular domain, and identification of its epitope should provide novel insight into the membrane topology of Cyt b. To that end, we examined biochemical features of 7D5-Cyt b binding and used the J404 phage display nonapeptide library to identify the bound epitope. 7D5 precipitated only heterodimeric gp91-p22phox and not individual or denatured Cyt b subunits from detergent extracts of human neutrophils and promyelocytic leukemia cells (gp91 PLB). Moreover, 7D5 precipitated precursor gp65-p22phox complexes from detergent extracts of the biosynthetically active gp91-PLB cells, demonstrating that complex carbohydrates were not required for epitope recognition. Epitope mimetics selected from the J404 phage display library by 7D5 demonstrated that (226)RIVRG(230) and (160)IKNP(163) regions of gp91phox were both bound by 7D5. These studies reveal specific information about Cyt b membrane topology and structure, namely that gp91phox residues (226)RIVRG(230) and (160)IKNP(163) are closely juxtaposed on extracytoplasmic domains and that predicted helices containing residues Gly(165)-Ile(190) and Ser(200)-Glu(225) are adjacent to each other in the membrane. PMID- 11027686 TI - Flavonoid 6-hydroxylase from soybean (Glycine max L.), a novel plant P-450 monooxygenase. AB - Cytochrome P-450-dependent hydroxylases are typical enzymes for the modification of basic flavonoid skeletons. We show in this study that CYP71D9 cDNA, previously isolated from elicitor-induced soybean (Glycine max L.) cells, codes for a protein with a novel hydroxylase activity. When heterologously expressed in yeast, this protein bound various flavonoids with high affinity (1.6 to 52 microm) and showed typical type I absorption spectra. These flavonoids were hydroxylated at position 6 of both resorcinol- and phloroglucinol-based A-rings. Flavonoid 6-hydroxylase (CYP71D9) catalyzed the conversion of flavanones more efficiently than flavones. Isoflavones were hardly hydroxylated. As soybean produces isoflavonoid constituents possessing 6,7-dihydroxy substitution patterns on ring A, the biosynthetic relationship of flavonoid 6-hydroxylase to isoflavonoid biosynthesis was investigated. Recombinant 2-hydroxyisoflavanone synthase (CYP93C1v2) efficiently used 6,7,4'-trihydroxyflavanone as substrate. For its structural identification, the chemically labile reaction product was converted to 6,7,4'-trihydroxyisoflavone by acid treatment. The structures of the final reaction products for both enzymes were confirmed by NMR and mass spectrometry. Our results strongly support the conclusion that, in soybean, the 6 hydroxylation of the A-ring occurs before the 1,2-aryl migration of the flavonoid B-ring during isoflavanone formation. This is the first identification of a flavonoid 6-hydroxylase cDNA from any plant species. PMID- 11027687 TI - Effect of heme iron valence state on the conformation of cytochrome c and its association with membrane interfaces. A CD and EPR investigation. AB - Recently cytochrome c has been mentioned as an important mediator in the events of cellular oxidative stress and apoptosis. To investigate the influence of charged interfaces on the conformation of cytochrome c, the CD and magnetic circular dichroic behavior of ferric and ferrous cytochrome c in homogeneous medium and in phosphatidylcholine/phosphatidylethanolamine/cardiolipin and dicetylphosphate liposomes was studied in the 300-600 and 200-320 nm wavelength region. EPR spectra demonstrate that the association of cytochrome c with membranes promotes alterations of the crystal field symmetry and spin state of the heme Fe(3+). The studies also include the effect of P(i), NaCl, and CaCl(2). Magnetic circular dichroism and CD results show that the interaction of both ferrous and ferric cytochrome c with charged interfaces promotes conformational changes in the alpha-helix content, tertiary structure, and heme iron spin state. Moreover, the association of cytochrome c with different liposomes is sensitive to the heme iron valence state. The more effective association with membranes occurs with ferrous cytochrome c. Dicetylphosphate liposomes, as a negatively charged membrane model, promoted a more pronounced conformational modification in the cytochrome c structure. A decrease in the lipid/protein association is detected in the presence of increasing amounts of CaCl(2), NaCl, and P(i), in response to the increase of the ionic strength. PMID- 11027688 TI - Identification and characterization of a Drosophila nuclear proteasome regulator. A homolog of human 11 S REGgamma (PA28gamma ). AB - We report the cloning and characterization of a Drosophila proteasome 11 S REGgamma (PA28) homolog. The 28-kDa protein shows 47% identity to the human REGgamma and strongly enhances the trypsin-like activities of both Drosophila and mammalian 20 S proteasomes. Surprisingly, the Drosophila REG was found to inhibit the proteasome's chymotrypsin-like activity against the fluorogenic peptide succinyl-LLVY-7-amino-4-methylcoumarin. Immunocytological analysis reveals that the Drosophila REG is localized to the nucleus but is distributed throughout the cell when nuclear envelope breakdown occurs during mitosis. Through site-directed mutagenesis studies, we have identified a functional nuclear localization signal present in the homolog-specific insert region. The Drosophila PA28 NLS is similar to the oncogene c-Myc nuclear localization motif. Comparison between uninduced and innate immune induced Drosophila cells suggests that the REGgamma proteasome activator has a role independent of the invertebrate immune system. Our results support the idea that gamma class proteasome activators have an ancient conserved function within metazoans and were present prior to the emergence of the alpha and beta REG classes. PMID- 11027689 TI - Structural and catalytic similarities between nucleotide pyrophosphatases/phosphodiesterases and alkaline phosphatases. AB - Nucleotide pyrophosphatases/phosphodiesterases (NPPs) generate nucleoside 5' monophosphates from a variety of nucleotides and their derivatives. Here we show by data base analysis that these enzymes are conserved from eubacteria to higher eukaryotes. We also provide evidence for the existence of two additional members of the mammalian family of ecto-NPPs. Homology searches and alignment-assisted mutagenesis revealed that the catalytic core of NPPs assumes a fold similar to that of a superfamily of phospho-/sulfo-coordinating metalloenzymes comprising alkaline phosphatases, phosphoglycerate mutases, and arysulfatases. Mutation of mouse NPP1 in some of its predicted metal-coordinating residues (D358N or H362Q) or in the catalytic site threonine (T238S) resulted in an enzyme that could still form the nucleotidylated catalytic intermediate but was hampered in the second step of catalysis. We also obtained data indicating that the ability of some mammalian NPPs to auto(de)phosphorylate is due to an intrinsic phosphatase activity, whereby the enzyme phosphorylated on Thr-238 represents the covalent intermediate of the phosphatase reaction. The results of site-directed mutagenesis suggested that the nucleotide pyrophosphatase/phosphodiesterase and the phosphatase activities of NPPs are mediated by a single catalytic site. PMID- 11027690 TI - Identification and structural characterization of nucleus-encoded transfer RNAs imported into wheat mitochondria. AB - Despite its large size (200-2400 kilobase pairs), the mitochondrial genome of angiosperms does not encode the minimal set of tRNAs required to support mitochondrial protein synthesis. Here we report the identification of cytosolic like tRNAs in wheat mitochondria using a method involving quantitative hybridization to distinguish among three tRNA classes: (i) those encoded by mitochondrial DNA (mtDNA) and localized in mitochondria, (ii) those encoded by nuclear DNA and located in the cytosol, and (iii) those encoded by nuclear DNA and found in both the cytosol and mitochondria. The latter class comprises tRNA species that are considered to be imported into mitochondria to compensate for the deficiency of mtDNA-encoded tRNAs. In a comprehensive survey of the wheat mitochondrial tRNA population, we identified 14 such imported tRNAs, the structural characterization of which is presented here. These imported tRNAs complement 16 mtDNA-encoded tRNAs, for a total of at least 30 distinct tRNA species in wheat mitochondria. Considering differences in the set of mtDNA encoded and imported tRNAs in the mitochondria of various land plants, the import system must be able to adapt relatively rapidly over evolutionary time with regard to the particular cytosolic-like tRNAs that are brought into mitochondria. PMID- 11027691 TI - Rate-limiting step preceding cytochrome c release in cells primed for Fas mediated apoptosis revealed by analysis of cellular mosaicism of respiratory changes. AB - In the present work, Jurkat cells undergoing anti-Fas antibody (anti-Fas) triggered apoptosis exhibited in increasing proportion a massive release of cytochrome c from mitochondria, as revealed by double-labeling confocal immunofluorescence microscopy. The cytochrome c release was followed by a progressive reduction in the respiratory activity of the last respiratory enzyme, cytochrome c oxidase (COX), and with a little delay, by a decrease in overall endogenous respiration rate, as measured in vivo in the whole cell population. Furthermore, in vivo titration experiments showed that an approximately 30% excess of COX capacity over that required to support endogenous respiration, found in naive cells, was maintained in anti-Fas-treated cells having lost approximately 40% of their COX respiratory activity. This observation strongly suggested that only a subpopulation of anti-Fas-treated cells, which maintained the excess of COX capacity, respired. Fractionation of cells on annexin V-coated paramagnetic beads did indeed separate a subpopulation of annexin V-binding apoptotic cells with fully released cytochrome c and completely lacking respiration, and a nonbound cell subpopulation exhibiting nearly intact respiration and in their great majority preserving the mitochondrial cytochrome c localization. The above findings showed a cellular mosaicism in cytochrome c release and respiration loss, and revealed the occurrence of a rate-limiting step preceding cytochrome c release in the apoptotic cascade. Furthermore, the striking observation that controlled digitonin treatment caused a massive and very rapid release of cytochrome c and complete loss of respiration in the still respiring anti-Fas-treated cells, but not in naive cells, indicated that the cells responding to digitonin had already been primed for apoptosis, and that this treatment bypassed or accelerated the rate-limiting step most probably at the level of the mitochondrial outer membrane. PMID- 11027692 TI - Relationship between novel isoforms, functionally important domains, and subcellular distribution of CD164/endolyn. AB - Functional analyses have indicated that the human CD164 sialomucin may play a key role in hematopoiesis by facilitating the adhesion of human CD34(+) cells to the stroma and by negatively regulating CD34(+)CD38(lo/-) cell proliferation. We have identified three novel human CD164 variants derived by alternative splicing of bona fide exons from a single genomic transcription unit. The predominant CD164(E1-6) isoform, encoded by six exons, is a type I transmembrane protein containing two extracellular mucin domains (I and II) interrupted by a cysteine rich non-mucin domain. The 103B2/9E10 and 105A5 epitopes, which specify ligand binding characteristics, are located on the exon 1-encoded mucin domain I. Three human CD164(E1-6) mRNA species, exhibiting differential polyadenylation site usage, are differentially expressed in hematopoietic and non-hematopoietic tissues. This study provides additional evidence that human CD164(E1-6) represents the ortholog of murine MGC-24v and rat endolyn. Comparative analysis of murine MGC-24v/CD164(E1-6) with human CD164(E1-6) revealed two potential splice variants and a similar genomic structure. Whereas the human CD164 gene is located on chromosome 6q21, the mouse gene occurs in a syntenic region on chromosome 10B1-B2. By confocal microscopy, human CD164 in CD34(+)CD38(+) hematopoietic progenitor (KG1B) and epithelial cell lines appears to be localized primarily in endosomes and lysosomes, with low concentrations at the cell surface. However, in a minority of KG1B cells, CD164 is more prominently expressed at the plasma membrane and in the recycling endosomes, suggesting that its distribution is regulated in cells of hematopoietic origin. PMID- 11027693 TI - The N-terminal N5 subdomain of the alpha 3(VI) chain is important for collagen VI microfibril formation. AB - Collagen VI assembly is unique within the collagen superfamily in that the alpha 1(VI), alpha 2(VI), and alpha 3(VI) chains associate intracellularly to form triple helical monomers, and then dimers and tetramers, which are secreted from the cell. Secreted tetramers associate end-to-end to form the distinctive extracellular microfibrils that are found in virtually all connective tissues. Although the precise protein interactions involved in this process are unknown, the N-terminal globular regions, which are composed of multiple copies of von Willebrand factor type A-like domains, are likely to play a critical role in microfibril formation, because they are exposed at both ends of the tetramers. To explore the role of these subdomains in collagen VI intracellular and extracellular assembly, alpha 3(VI) cDNA expression constructs with sequential N terminal deletions were stably transfected into SaOS-2 cells, producing cell lines that express alpha 3(VI) chains with N-terminal globular domains containing modules N9-N1, N6-N1, N5-N1, N4-N1, N3-N1, or N1, as well as the complete triple helix and C-terminal globular domain (C1-C5). All of these transfected alpha 3(VI) chains were able to associate with endogenous alpha 1(VI) and alpha 2(VI) to form collagen VI monomers, dimers, and tetramers, which were secreted. Importantly, cells that expressed alpha 3(VI) chains containing the N5 subdomain, alpha 3(VI) N9-C5, N6-C5, and N5-C5, formed microfibrils and deposited a collagen VI matrix. In contrast, cells that expressed the shorter alpha 3(VI) chains, N4 C5, N3-C5, and N1-C5, were severely compromised in their ability to form end-to end tetramer assemblies and failed to deposit a collagen VI matrix. These data demonstrate that the alpha 3(VI) N5 module is critical for microfibril formation, thus identifying a functional role for a specific type A subdomain in collagen VI assembly. PMID- 11027694 TI - The RihA, RihB, and RihC ribonucleoside hydrolases of Escherichia coli. Substrate specificity, gene expression, and regulation. AB - Pyrimidine-requiring cdd mutants of Escherichia coli deficient in cytidine deaminase utilize cytidine as a pyrimidine source by an alternative pathway. This has been presumed to involve phosphorylation of cytidine to CMP by cytidine/uridine kinase and subsequent hydrolysis of CMP to cytosine and ribose 5 phosphate by a putative CMP hydrolase. Here we show that cytidine, in cdd strains, is converted directly to cytosine and ribose by a ribonucleoside hydrolase encoded by the previously uncharacterized gene ybeK, which we have renamed rihA. The RihA enzyme is homologous to the products of two unlinked genes, yeiK and yaaF, which have been renamed rihB and rihC, respectively. The RihB enzyme was shown to be a pyrimidine-specific ribonucleoside hydrolase like RihA, whereas RihC hydrolyzed both pyrimidine and purine ribonucleosides. The physiological function of the ribonucleoside hydrolases in wild-type E. coli strains is enigmatic, as their activities are paralleled by the phosphorolytic activities of the nucleoside phosphorylases, and a triple mutant lacking all three hydrolytic activities grew normally. Furthermore, enzyme assays and lacZ gene fusion analysis indicated that rihB was essentially silent unless activated by mutation, whereas rihA and rihC were poorly expressed in glucose medium due to catabolite repression. PMID- 11027695 TI - Formation of the covalent chymotrypsin.antichymotrypsin complex involves no large scale movement of the enzyme. AB - alpha(1)-Antichymotrypsin is a member of the serine proteinase inhibitor, or serpin, family that typically forms very long-lived, enzymatically inactive 1:1 complexes (denoted E*I*) with its target proteinases. Serpins share a conserved tertiary structure, in which an exposed region of amino acid residues (called the reactive center loop or RCL) acts as bait for a target proteinase. Within E*I*, the two proteins are linked covalently as a result of nucleophilic attack by Ser(195) of the serine proteinase on the P1 residue within the RCL of the serpin. This species is formally similar to the acyl enzyme species normally seen as an intermediate in serpin proteinase catalysis. However, its subsequent hydrolysis is extremely slow as a result of structural changes within the enzyme leading to distortion of the active site. There is at present an ongoing debate concerning the structure of the E*I* complex; in particular, as to whether the enzyme, bound to P1, maintains its original position at the top of the serpin molecule or instead translocates across the entire length of the serpin, with concomitant insertion of RCL residues P1-P14 within beta-sheet A and a large separation of the enzyme and RCL residue P1'. We report time-resolved fluorescence energy transfer and rapid mixing/quench studies that support the former model. Our results indicate that the distance between residue P1' in alpha(1) antichymotrypsin and the amino terminus of chymotrypsin actually decreases on conversion of the encounter complex E.I to E*I*. These results led us to formulate a comprehensive mechanism that accounted both for our results and for those of others supporting the two different E*I* structures. In this mechanism, partial insertion of the RCL, with no large perturbation of the P1' enzyme distance, is followed by covalent acyl enzyme formation. Full insertion can subsequently take place, in a reversible fashion, with the position of equilibrium between the partially and fully inserted complexes depending on the particular serpin-proteinase pair under consideration. PMID- 11027696 TI - Molecular cloning, chromosomal localization, tissue mRNA levels, bacterial expression, and enzymatic properties of human NMN adenylyltransferase. AB - A 1329-base pair clone isolated from a human placenta cDNA library contains a full-length 837-base pair coding region for a 31.9-kDa protein whose deduced primary structure exhibits high homology to consensus sequences found in other NMN adenylyltransferases. Northern blotting detected a major 3.1-kilobase mRNA transcript as well as a minor 4.1-kilobase transcript in all human tissues examined. In several cancer cell lines, lower levels of mRNA expression were clearly evident. The gene encoding the human enzyme was mapped to chromosome band 1p32-35. High efficiency bacterial expression yielded 1.5 mg of recombinant enzyme/liter of culture medium. The molecular and kinetic properties of recombinant human NMN adenylyltransferase provide new directions for investigating metabolic pathways involving this enzyme. PMID- 11027697 TI - Ending world hunger. The promise of biotechnology and the threat of antiscience zealotry. PMID- 11027698 TI - Several features of the GT-factor trihelix domain resemble those of the Myb DNA binding domain. PMID- 11027699 TI - The cellulose synthase superfamily. PMID- 11027700 TI - Plant Signaling 2000. Cross talk among geneticists, physiologists, and ecologists. AB - Plants respond in complex ways to their environment, to their internal physiological status, and to the activity of other plants, pathogens, herbivores, and organisms. Plant Signaling 2000, a symposium sponsored by the Penn State Intercollege Graduate Program in Plant Physiology (May 18-20, 2000), explored the machinery underlying these responses and their potential for cross talk. We recount here some of the major themes emerging from this interdisciplinary symposium, which ranged from genetic and biochemical analyses of signaling pathways in Arabidopsis and other model plants to field studies of plants responding to insect damage. PMID- 11027701 TI - Medicinal plants and phytomedicines. Linking plant biochemistry and physiology to human health. PMID- 11027702 TI - Do legume storage proteins play a role in defending seeds against bruchids? PMID- 11027703 TI - Insights into a key developmental switch and its importance for efficient plant breeding. PMID- 11027704 TI - Regulators and regulation of legume root nodule development. PMID- 11027705 TI - Clausa, a tomato mutant with a wide range of phenotypic perturbations, displays a cell type-dependent expression of the homeobox gene LeT6/TKn2. AB - Class I knox genes play an important role in shoot meristem function and are thus involved in the ordered development of stems, leaves, and reproductive organs. To elucidate the mechanism underlying the expression pattern of these homeobox genes, we studied a spontaneous tomato (Lycopersicon esculentum) mutant that phenotypically resembles, though is more extreme than, transgenic plants misexpressing class I knox genes. This mutant was found to carry a recessive allele, denoted clausa:shootyleaf (clau:shl)-a newly identified allele of clausa. Mutant plants exhibited abnormal leaf and flower morphology, epiphyllus inflorescences, fusion of organs, calyx asymmetry, and navel-like fruits. Analysis by scanning electron microscopy revealed that such fruits carried ectopic ovules, various vegetative primordia, as well as "forests" of stalked glandular trichomes. In situ RNA hybridization showed a peculiar expression pattern of the class I knox gene LeT6/TKn2; expression was restricted to the vascular system and palisade layer of mature leaves and to the inner part of ovules integuments. We conclude that CLAUSA regulates various aspects of tomato plant development, at least partly, by rendering the LeT6/TKn2 gene silent in specific tissues during development. Considering the expression pattern of LeT6/TKn2 in the clausa mutant, we suggest that the control over a given homeobox gene is maintained by several different regulatory mechanisms, in a cell type dependent manner. PMID- 11027706 TI - Hormonal interactions in the control of Arabidopsis hypocotyl elongation. AB - The Arabidopsis hypocotyl, together with hormone mutants and chemical inhibitors, was used to study the role of auxin in cell elongation and its possible interactions with ethylene and gibberellin. When wild-type Arabidopsis seedlings were grown on media containing a range of auxin concentrations, hypocotyl growth was inhibited. However, when axr1-12 and 35S-iaaL (which have reduced auxin response and levels, respectively) were grown in the same conditions, auxin was able to promote hypocotyl growth. In contrast, auxin does not promote hypocotyl growth of axr3-1, which has phenotypes that suggest an enhanced auxin response. These results are consistent with the hypothesis that auxin levels in the wild type hypocotyl are optimal for elongation and that additional auxin is inhibitory. When ethylene responses were reduced using either the ethylene resistant mutant etr1 or aminoethoxyvinylglycine, an inhibitor of ethylene synthesis, auxin responses were unchanged, indicating that auxin does not inhibit hypocotyl elongation through ethylene. To test for interactions between auxin and gibberellin, auxin mutants were grown on media containing gibberellin and gibberellin mutants were grown on media containing auxin. The responses were found to be the same as wild-type Arabidopsis seedlings in all cases. In addition, 1 microM of the auxin transport inhibitor 1-naphthylphthalmic acid does not alter the response of wild-type seedlings to gibberellin. Double mutants were made between gibberellin and auxin mutants and the phenotypes of these appear additive. These results indicate that auxin and gibberellin are acting independently in hypocotyl elongation. Thus auxin, ethylene, and gibberellin each regulate hypocotyl elongation independently. PMID- 11027707 TI - Essential role of caffeoyl coenzyme A O-methyltransferase in lignin biosynthesis in woody poplar plants. AB - Caffeoyl coenzyme A O-methyltransferase (CCoAOMT) has recently been shown to participate in lignin biosynthesis in herbacious tobacco plants. Here, we demonstrate that CCoAOMT is essential in lignin biosynthesis in woody poplar (Populus tremula x Populus alba) plants. In poplar stems, CCoAOMT was found to be expressed in all lignifying cells including vessel elements and fibers as well as in xylem ray parenchyma cells. Repression of CCoAOMT expression by the antisense approach in transgenic poplar plants caused a significant decrease in total lignin content as detected by both Klason lignin assay and Fourier-transform infrared spectroscopy. The reduction in lignin content was the result of a decrease in both guaiacyl and syringyl lignins as determined by in-source pyrolysis mass spectrometry. Fourier-transform infrared spectroscopy indicated that the reduction in lignin content resulted in a less condensed and less cross linked lignin structure in wood. Repression of CCoAOMT expression also led to coloration of wood and an elevation of wall-bound p-hydroxybenzoic acid. Taken together, these results indicate that CCoAOMT plays a dominant role in the methylation of the 3-hydroxyl group of caffeoyl CoA, and the CCoAOMT-mediated methylation reaction is essential to channel substrates for 5-methoxylation of hydroxycinnamates. They also suggest that antisense repression of CCoAOMT is an efficient means for genetic engineering of trees with low lignin content. PMID- 11027708 TI - The Arabidopsis UVH1 gene is a homolog of the yeast repair endonuclease RAD1. AB - Ultraviolet radiation induces DNA damage products, largely in the form of pyrimidine dimers, that are both toxic and mutagenic. In most organisms, including Arabidopsis, these lesions are repaired both through a dimer-specific photoreactivation mechanism and through a less efficient light-independent mechanism. Several mutants defective in this "dark repair" pathway have been previously described. The mechanism of this repair has not been elucidated, but is thought to be homologous to the nucleotide excision repair mechanisms found in other eukaryotes. Here we report the complementation of the Arabidopsis uvh1 dark repair mutant with the Arabidopsis homolog of the yeast nucleotide excision repair gene RAD1, which encodes one of the subunits of the 5'-repair endonuclease. The uvh1-2 mutant allele carries a glycine-->aspartate amino acid change that has been previously identified to produce a null allele of RAD1 in yeast. Although Arabidopsis homologs of genes involved in nucleotide excision repair are readily identified by searching the genomic database, it has not been established that these homologs are actually required for dark repair in plants. The complementation of the Arabidopsis uvh1 mutation with the Arabidopsis RAD1 homolog clearly demonstrates that the mechanism of nucleotide excision repair is conserved among the plant, animal, and fungal kingdoms. PMID- 11027709 TI - Expression of a gene encoding mitochondrial aldehyde dehydrogenase in rice increases under submerged conditions. AB - It is known that alcoholic fermentation is important for survival of plants under anaerobic conditions. Acetaldehyde, one of the intermediates of alcoholic fermentation, is not only reduced by alcohol dehydrogenase but also can be oxidized by aldehyde dehydrogenase (ALDH). To determine whether ALDH plays a role in anaerobic metabolism in rice (Oryza sativa L. cv Nipponbare), we characterized a cDNA clone encoding mitochondrial ALDH from rice (Aldh2a). Analysis of sub cellular localization of ALDH2a protein using green fluorescent protein and an in vitro ALDH assay using protein extracts from Escherichia coli cells that overexpressed ALDH2a indicated that ALDH2a functions in the oxidation of acetaldehyde in mitochondria. A Southern-blot analysis indicated that mitochondrial ALDH is encoded by at least two genes in rice. We found that the Aldh2a mRNA was present at high levels in leaves of dark-grown seedlings, mature leaf sheaths, and panicles. It is interesting that expression of the rice Aldh2a gene, unlike the expression of the tobacco (Nicotiana tabacum) Aldh2a gene, was induced in rice seedlings by submergence. Experiments with ruthenium red, which is a blocker of Ca(2+) fluxes in rice as well as maize (Zea mays), suggest that the induction of expression of Adh1 and Pdc1 by low oxygen stress is regulated by elevation of the cytosolic Ca(2+) level. However, the induction of Aldh2a gene expression may not be controlled by the cytosolic Ca(2+) level elevation. A possible involvement of ALDH2a in the submergence tolerance of rice is discussed. PMID- 11027710 TI - Identification of a new glucosinolate-rich cell type in Arabidopsis flower stalk. AB - Distribution of K, Ca, Cl, S, and P in freeze-dried sections of Arabidopsis flower stalk was analyzed by energy dispersive x-ray imaging. Concentrations of these elements in different cell types were quantified by microanalysis of single cell samples and phloem exudates. Results showed a differential pattern of distribution for all five elements. K concentration was found to be highest in the parenchymatous tissue around vascular bundles. Ca and Cl were present mainly in the central part of the flower stalk. P was largely located in the bundles and in the parenchyma surrounding them. S signal was extraordinary high in groups of cells (S-cells) situated between the phloem of every vascular bundle and the endodermis. Enzymatic hydrolysis by thioglucosidase of cell sap collected from S cells using a glass microcapillary resulted in the release of glucose, indicating that these cells contain glucosinolates at high (> 100 mM) concentration, which is consistent with the concentration of S (> 200 mM) estimated by x-ray analysis of cell sap samples. Since their position outside of the phloem is ideally suited for protecting the long-distance transport system from feeding insects, the possible roles of these cells as components of a plant defense system are discussed. PMID- 11027711 TI - Ethylene induces epidermal cell death at the site of adventitious root emergence in rice. AB - In deepwater rice (Oryza sativa), adventitious root primordia initiate at the nodes as part of normal development. Emergence of the roots is dependent on flooding of the plant and is mediated by ethylene action. Root growth was preceded by the induced death of epidermal cells of the node external to the tip of the root primordium. Cell death proceeded until the epidermis split open. Through this crack the root eventually emerged. Induced death was confined to nodal epidermal cells covering the tip of the primordia. Our results suggest that this process facilitates adventitious root emergence and prevents injury to the growing root. Cell death was inducible not only by submergence but also by application of 1-aminocyclopropane-1-carboxylic acid, the natural precursor of ethylene and it was suppressed in the presence of 2,5-norbornadiene (bicyclo[2.2.1]hepta-2,5-diene), an inhibitor of ethylene action. Adventitious root growth and epidermal cell death are therefore linked to the ethylene signaling pathway, which is activated in response to low oxygen stress. PMID- 11027712 TI - Characterization of Arabidopsis acid phosphatase promoter and regulation of acid phosphatase expression. AB - The expression and secretion of acid phosphatase (APase) was investigated in Indian mustard (Brassica juncea L. Czern.) plants using sensitive in vitro and activity gel assays. Phosphorus (P) starvation induced two APases in Indian mustard roots, only one of which was secreted. Northern-blot analysis indicated transcriptional regulation of APase expression. Polymerase chain reaction and Southern-blot analyses revealed two APase homologs in Indian mustard, whereas in Arabidopsis, only one APase homolog was detected. The Arabidopsis APase promoter region was cloned and fused to the beta-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes. GUS expression was first evident in leaves of the P starved Arabidopsis plants. In P-starved roots, the expression of GUS initiated in lateral root meristems followed by generalized expression throughout the root. GUS expression diminished with the addition of P to the medium. Expression of GFP in P-starved roots also initiated in the lateral root meristems and the recombinant GFP with the APase signal peptide was secreted by the roots into the medium. The APase promoter was specifically activated by low P levels. The removal of other essential elements or the addition of salicylic or jasmonic acids, known inducers of gene expression, did not activate the APase promoter. This novel APase promoter may be used as a plant-inducible gene expression system for the production of recombinant proteins and as a tool to study P metabolism in plants. PMID- 11027713 TI - A DEFICIENS homolog from the dioecious tree black cottonwood is expressed in female and male floral meristems of the two-whorled, unisexual flowers. AB - We isolated PTD, a member of the DEFICIENS (DEF) family of MADS box transcription factors, from the dioecious tree, black cottonwood (Populus trichocarpa). In females, in situ hybridization experiments showed that PTD mRNA was first detectable in cells on the flanks of the inflorescence meristem, before differentiation of individual flowers was visually detectable. In males, the onset of PTD expression was delayed until after individual flower differentiation had begun and floral meristems were developing. Although PTD was initially expressed throughout the inner whorl meristem in female and male flowers, its spatial expression pattern became sex-specific as reproductive primordia began to form. PTD expression was maintained in stamen primordia, but excluded from carpel primordia, as well as vegetative tissues. Although PTD is phylogenetically most closely related to the largely uncharacterized TM6 subfamily of the DEF/APETELA3(AP3)/TM6 group, its spatio-temporal expression patterns are more similar to that of DEF and AP3 than to other members of the TM6 subfamily. PMID- 11027715 TI - Distribution of peltate glandular trichomes on developing leaves of peppermint. AB - The pattern of peltate glandular trichome initiation and ontogeny on expanding peppermint (Mentha x piperita) leaves was defined by surveying the populations of peltate glands in each of seven developmental stages within sampling areas of leaf apical, mid-, and basal zones for both abaxial and adaxial surfaces. It was shown that new peltate glands continue to form until leaf expansion ceases and that regions of active gland initiation are unevenly distributed. The distribution of gland initiation reflects the basipetal pattern of leaf maturation, with relatively immature regions at the leaf base continuing to produce oil glands long after gland production has stopped at the leaf apex. The proportion of glands in the secretory stage as a function of leaf development and the direct observations of living glands over a period of 33 h indicate that a period of only 20 to 30 h of secretory activity is required for filling of the gland storage compartment with essential oil. These findings are discussed in relation to earlier literature describing age-related changes in glandular essential oil content. PMID- 11027714 TI - Tissue-specific and developmental pattern of expression of the rice sps1 gene. AB - Sucrose-phosphate synthase (SPS) is one of the key regulatory enzymes in carbon assimilation and partitioning in plants. SPS plays a central role in the production of sucrose in photosynthetic cells and in the conversion of starch or fatty acids into sucrose in germinating seeds. To explore the mechanisms that regulate the tissue-specific and developmental distribution of SPS, the expression pattern of rice (Oryza sativa) sps1 (GenBank accession no. U33175) was examined by in situ reverse transcriptase-polymerase chain reaction and the expression directed by the sps1 promoter using the beta-glucuronidase reporter gene. It was found that the expression of the rice sps1 gene is limited to mesophyll cells in leaves, the scutellum of germinating seedlings, and pollen of immature inflorescences. During leaf development, the sps1 promoter directs a basipetal pattern of expression that coincides with the distribution of SPS activity during the leaf sink-to-source transition. It was also found that during the vegetative part of the growth cycle, SPS expression and enzymatic activity are highest in the youngest fully expanded leaf. Additionally, it was observed that the expression of the sps1 promoter is regulated by light and dependent on plastid development in photosynthetic tissues, whereas expression in scutellum is independent of both light and plastid development. PMID- 11027716 TI - Development of peltate glandular trichomes of peppermint. AB - Cryofixation and conventional chemical fixation methods were employed to examine the ultrastructure of developing peltate glandular trichomes of peppermint (Mentha x piperita). Our results are discussed in relation to monoterpene production and the mechanism of essential oil secretion. Peltate glands arise as epidermal protuberances (initials) that divide asymmetrically to produce a vacuolate basal cell, a stalk cell, and a cytoplasmically dense apical cell. Further divisions of the apical cell produce a peltate trichome with one basal cell, one stalk cell, and eight glandular (secretory) disc cells. Presecretory gland cells resemble meristematic cells because they contain proplastids, small vacuoles, and large nuclei. The secretory phase coincides with the separation and filling of the sub-cuticular oil storage space, the maturation of glandular disc cell leucoplasts in which monoterpene biosynthesis is known to be initiated, and the formation of extensive smooth endoplasmic reticulum at which hydroxylation steps of the monoterpene biosynthetic pathway occur. The smooth endoplasmic reticulum of the secretory cells appears to form associations with both the leucoplasts and the plasma membrane bordering the sub-cuticular oil storage cavity, often contains densely staining material, and may be involved with the transport of the monoterpene-rich secretion product. Associated changes in the ultrastructure of the secretory stage stalk cell are also described, as is the ultrastructure of the fragile post-secretory gland for which cryofixation methods are particularly well suited for the preservation of organizational integrity. PMID- 11027718 TI - Abscisic acid stimulation of phospholipase D in the barley aleurone is G-protein mediated and localized to the plasma membrane. AB - We have previously determined that phospholipase D (PLD) is activated by abscisic acid (ABA), and this activation is required for the ABA response of the cereal aleurone cell. In this study, ABA-stimulated PLD activity was reconstituted in vitro in microsomal membranes prepared from aleurone protoplasts. The transient nature (20 min) and degree (1.5- to 2-fold) of activation in vitro were similar to that measured in vivo. Stimulation by ABA was only apparent in the membrane fraction and was associated with a fraction enriched in plasma membrane. These results suggest that an ABA receptor system and elements linking it to PLD activation are associated with the aleurone plasma membrane. The activation of PLD in vitro by ABA was dependent on the presence of GTP. Addition of GTPgammaS transiently stimulated PLD in an ABA-independent manner, whereas treatment with GDPbetaS or pertussis toxin blocked the PLD activation by ABA. Application of pertussis toxin to intact aleurone protoplasts inhibited the ability of ABA to activate PLD as well as antagonizing the ability of ABA to down-regulate gibberellic acid-stimulated alpha-amylase production. All of these data support the hypothesis that ABA stimulation of PLD activity occurs at the plasma membrane and is mediated by G-protein activity. PMID- 11027717 TI - Stearoyl-acyl carrier protein and unusual acyl-acyl carrier protein desaturase activities are differentially influenced by ferredoxin. AB - Acyl-acyl carrier protein (ACP) desaturases function to position a single double bond into an acyl-ACP substrate and are best represented by the ubiquitous Delta9 18:0-ACP desaturase. Several variant acyl-ACP desaturases have also been identified from species that produce unusual monoenoic fatty acids. All known acyl-ACP desaturase enzymes use ferredoxin as the electron-donating cofactor, and in almost all previous studies the photosynthetic form of ferredoxin rather than the non-photosynthetic form has been used to assess activity. We have examined the influence of different forms of ferredoxin on acyl-ACP desaturases. Using combinations of in vitro acyl-ACP desaturase assays and [(14)C]malonyl-coenzyme A labeling studies, we have determined that heterotrophic ferredoxin isoforms support up to 20-fold higher unusual acyl-ACP desaturase activity in coriander (Coriandrum sativum), Thunbergia alata, and garden geranium (Pelargonium x hortorum) when compared with photosynthetic ferredoxin isoforms. Heterotrophic ferredoxin also increases activity of the ubiquitous Delta9 18:0-ACP desaturase 1.5- to 3.0-fold in both seed and leaf extracts. These results suggest that ferredoxin isoforms may specifically interact with acyl-ACP desaturases to achieve optimal enzyme activity and that heterotrophic isoforms of ferredoxin may be the in vivo electron donor for this reaction. PMID- 11027719 TI - Redox-regulated RNA helicase expression. AB - In photosynthetic organisms it is becoming increasingly evident that light-driven shifts in redox potential act as a sensor that initiates alterations in gene expression at both the level of transcription and translation. This report provides evidence that the expression of a cyanobacterial RNA helicase gene, crhR, is controlled at the level of transcription and mRNA stability by a complex series of interacting mechanisms that are redox regulated. Transcript accumulation correlates with reduction of the electron transport chain between Q(A) in photosystem II and Q(O) in cyt b(6)f, when Synechocystis sp. strain PCC 6803 is cultured photoautotrophically or photomixotrophically and subjected to darkness and/or electron transport inhibitors or illumination that preferentially excites photosystem II. crhR mRNA stability is also regulated by a redox responsive mechanism, which differs from that affecting accumulation and does not involve signaling initiated by photoreceptors. The data are most consistent with plastoquinol/cyt b(6)f interaction as the sensor initiating a signal transduction cascade resulting in accumulation of the crhR transcript. Functionally, CrhR RNA unwinding could act as a linker between redox regulated transcription and translation. The potential for translational regulation of redox-induced gene expression through RNA helicase-catalyzed modulation of RNA secondary structure is discussed. PMID- 11027720 TI - Differential subcellular localization and expression of ATP sulfurylase and 5' adenylylsulfate reductase during ontogenesis of Arabidopsis leaves indicates that cytosolic and plastid forms of ATP sulfurylase may have specialized functions. AB - ATP sulfurylase and 5'-adenylylsulfate (APS) reductase catalyze two reactions in the sulfate assimilation pathway. Cell fractionation of Arabidopsis leaves revealed that ATP sulfurylase isoenzymes exist in the chloroplast and the cytosol, whereas APS reductase is localized exclusively in chloroplasts. During development of Arabidopsis plants the total activity of ATP sulfurylase and APS reductase declines by 3-fold in leaves. The decline in APS reductase can be attributed to a reduction of enzyme during aging of individual leaves, the highest activity occurring in the youngest leaves and the lowest in fully expanded leaves. By contrast, total ATP sulfurylase activity declines proportionally in all the leaves. The distinct behavior of ATP sulfurylase can be attributed to reciprocal expression of the chloroplast and cytosolic isoenzymes. The chloroplast form, representing the more abundant isoenzyme, declines in parallel with APS reductase during aging; however, the cytosolic form increases over the same period. In total, the results suggest that cytosolic ATP sulfurylase plays a specialized function that is probably unrelated to sulfate reduction. A plausible function could be in generating APS for sulfation reactions. PMID- 11027721 TI - Rac-related GTP-binding protein in elicitor-induced reactive oxygen generation by suspension-cultured soybean cells. AB - Plant cells produce reactive oxygen species (ROS) in response to many stimuli. However, the mechanism of ROS biosynthesis remains unclear. We have explored the hypothesis that the superoxide burst in plants mechanistically resembles the oxidative burst in neutrophils. First we have confirmed that ROS production, which occurs in suspension-cultured soybean (Glycine max) cells in response to hypo-osmotic shock, is inhibited by diphenylene iodonium, an inhibitor of the flavin-dependent oxidase of neutrophils. Because a Rac family G protein is an essential regulator of this NADPH oxidase, and because many plant homologs of Rac have been cloned, we next examined whether Rac-like proteins might be involved in the oxidative burst in the soybean cells. We identified a Rac-like 21-kD soybean protein that cross-reacts with antibodies to human Rac and garden pea Rop and also binds [gamma-(35)S] GTP, a diagnostic trait of small G proteins. This Rac related protein translocated from the cytosol to microsomes during the oxidative burst. Moreover, soybean cells transiently transformed with either a dominant negative (RacN17) or a dominant positive (RacV12) form of Rac1 showed the anticipated altered responses to three different stimuli: hypo-osmotic shock, oligo-GalUA, and harpin. In response to these stimuli, cells transformed with RacN17 produced less ROS and cells transformed with RacV12 generated more ROS than control cells. These results strongly suggest that a Rac-related protein participates in the regulation of ROS production in soybean cells, possibly via activation of an enzyme complex similar to the NADPH oxidase of phagocytes in animal systems. PMID- 11027722 TI - Nod factors and chitooligomers elicit an increase in cytosolic calcium in aequorin-expressing soybean cells. AB - Rhizobial Nod factors (NFs) function as nodulation signals that trigger symbiotic responses of leguminous host plants. NFs consist of a chitin oligomer backbone carrying a fatty acid at the non-reducing end. Depending on the rhizobial strain, NFs carry additional substituents, which may determine host specificity. Transgenic suspension-cultured soybean (Glycine max [L.] Merr.) cells expressing aequorin have been used to record cytosolic [Ca(2+)] changes upon treatment with purified NFs and chitin fragments. Both compounds elicited an increase of cytosolic [Ca(2+)] at nanomolar concentrations. The shape and amplitude of cytosolic [Ca(2+)] changes was similar to the response elicited by un-derivatized chitin oligomers. Cells challenged first with NFs did not respond to a subsequent treatment with chitin oligomers and vice versa. Dose-response experiments showed that un-derivatized chitin oligomers were more active compared with NFs. The capacity of NFs to elicit the calcium response depended on their structure. The presence of reducing end substituents in methylfucosylated NFs from Rhizobium sp. NGR234 and the O-acetyl group at the non-reducing end in NFs from Sinorhizobium meliloti attenuated the activity to cause the calcium changes. The sulfate group in NFs from Rhizobium tropici did not affect the elicitor activity. Pentameric S. meliloti NFs were more active than tetrameric molecules, whereas trimeric or dimeric degradation products were inactive. Substituents in NFs may have the function to avoid stimulation of defense reactions mediated by the perception system for chitin oligomers. PMID- 11027723 TI - Accumulation of ENOD2-like transcripts in non-nodulating woody papilionoid legumes. AB - Japanese pagodatree (Styphnolobium japonicum [L.] Schott) and American yellowwood (Cladrastis kentukea Dum.-Cours.) Rudd are the first woody, non-nodulating papilionoid legumes shown to possess putative early nodulin 2 (ENOD2) genes. ENOD2 cDNAs from Japanese pagodatree (807 bp) and American yellowwood (735 bp) have 75% to 79% sequence identity to ENOD2 sequences and encode deduced proteins that possess conserved ENOD2 pentapeptides (PPHEK and PPEYQ). Lower percentages of glucose and higher percentages of histidine and valine suggest that SjENOD2 and CkENOD2 are different from other ENOD2s. Hybridization analyses indicate the clones represent ENOD2 gene families of two to four genes in Japanese pagodatree and American yellowwood genomes, and ENOD2-like transcripts were detected in stems and flowers, as well as roots. Only roots of control species that nodulate, Maackia amurensis Rupr. & Maxim. and alfalfa (Medicago sativa), produced pseudonodules after treatment with zeatin or 2,3,5-triiodobenzoic acid, an auxin transport inhibitor. Accumulation of MaENOD2 transcripts was enhanced during the first 10 d of treatment, but 2,3,5-triiodobenzoic acid and zeatin enhanced transcript accumulation after 30 d in roots of Japanese pagodatree and American yellowwood. Characteristics that distinguish ENOD2 gene families in basal, non nodulating woody legumes from other ENOD2 genes may provide new information about the function of these genes during symbiotic and non-symbiotic organ development. PMID- 11027724 TI - Recombinant brassinosteroid insensitive 1 receptor-like kinase autophosphorylates on serine and threonine residues and phosphorylates a conserved peptide motif in vitro. AB - BRASSINOSTEROID-INSENSITIVE 1 (BRI1) encodes a putative Leucine-rich repeat receptor kinase in Arabidopsis that has been shown by genetic and molecular analysis to be a critical component of brassinosteroid signal transduction. In this study we examined some of the biochemical properties of the BRI1 kinase domain (BRI1-KD) in vitro, which might be important predictors of in vivo function. Recombinant BRI1-KD autophosphorylated on serine (Ser) and threonine (Thr) residues with p-Ser predominating. Matrix-assisted laser desorption/ionization mass spectrometry identified a minimum of 12 sites of autophosphorylation in the cytoplasmic domain of BRI1, including five in the juxtamembrane region (N-terminal to the catalytic KD), five in the KD (one each in sub-domains I and VIa and three in sub-domain VIII), and two in the carboxy terminal region. Five of the sites were uniquely identified (Ser-838, Thr-842, Thr-846, Ser-858, and Thr-872), whereas seven were localized on short peptides but remain ambiguous due to multiple Ser and/or Thr residues within these peptides. The inability of an active BRI1-KD to transphosphorylate an inactive mutant KD suggests that the mechanism of autophosphorylation is intramolecular. It is interesting that recombinant BRI1-KD was also found to phosphorylate certain synthetic peptides in vitro. To identify possible structural elements required for substrate recognition by BRI1-KD, a series of synthetic peptides were evaluated, indicating that optimum phosphorylation of the peptide required R or K residues at P - 3, P - 4, and P + 5 (relative to the phosphorylated Ser at P = 0). PMID- 11027725 TI - Effects of elevated [CO(2)] and nitrogen nutrition on cytokinins in the xylem sap and leaves of cotton. AB - We measured the level of xylem-derived cytokinins (CKs) entering a cotton leaf, and the CK levels in the same leaf, thus enabling xylem sap and foliar CKs to be compared concurrently. Although zeatin was the dominant CK in xylem sap, zeatin, dihydrozeatin, and N(6)-(2-isopentenyl) adenine were present in approximately equimolar levels in leaves. Elevated [CO(2)] (EC) has an effect on the levels of cytokinins in sap and leaf tissues. This effect was modulated by the two levels of root nitrogen nutrition (2 and 12 mM nitrate). Growth enhancement (70%) in EC over plants in ambient [CO(2)] (AC) was observed for both nitrogen nutrition treatments. Low-nitrogen leaves growing in EC exhibited photosynthetic acclimation, whereas there was no sign of photosynthetic acclimation in high nitrogen grown leaves. Under these prevailing conditions, xylem sap and leaf tissues were obtained for CK analysis. Higher nitrogen nutrition increased the delivery per unit leaf area of CKs to the leaf at AC. EC caused a greater increase in CK delivery to the leaf at low nitrogen conditions (106%) than at high nitrogen conditions (17%). EC induced a significant increase in CK content in low-nitrogen leaves, whereas CK content in leaf tissues was similar for high nitrogen leaves growing in AC and EC. PMID- 11027726 TI - Production of the isoflavones genistein and daidzein in non-legume dicot and monocot tissues. AB - Metabolic engineering for production of isoflavones in non-legume plants may provide the health benefits of these phytoestrogens from consumption of more widely used grains. In legumes, isoflavones function in both the symbiotic relationship with rhizobial bacteria and the plant defense response. Expression of a soybean isoflavone synthase (IFS) gene in Arabidopsis plants was previously shown to result in the synthesis and accumulation of the isoflavone genistein in leaf and stem tissue (Jung et al., 2000). Here we further investigate the ability of the heterologous IFS enzyme to interact with the endogenous phenylpropanoid pathway, which provides the substrate for IFS, and produces genistein in several plant tissue systems. In tobacco (Nicotiana tabacum) floral tissue that synthesizes anthocyanins, genistein production was increased relative to leaves. Induction of the flavonoid/anthocyanin branch of the phenylpropanoid pathway through UV-B treatment also enhanced genistein production in Arabidopsis. In a monocot cell system, introduced expression of a transcription factor regulating genes of the anthocyanin pathway was effective in conferring the ability to produce genistein in the presence of the IFS gene. Introduction of a third gene, chalcone reductase, provided the ability to synthesize an additional substrate of IFS resulting in production of the isoflavone daidzein in this system. The genistein produced in tobacco, Arabidopsis, and maize (Zea mays) cells was present in conjugated forms, indicating that endogenous enzymes were capable of recognizing genistein as a substrate. This study provides insight into requirements for metabolic engineering for isoflavone production in non-legume dicot and monocot tissues. PMID- 11027728 TI - Changes in gibberellin A(1) levels and response during de-etiolation of pea seedlings. AB - The level of gibberellin A(1) (GA(1)) in shoots of pea (Pisum sativum) dropped rapidly during the first 24 h of de-etiolation. The level then increased between 1 and 5 d after transfer to white light. Comparison of the metabolism of [(13)C(3)H] GA(20) suggested that the initial drop in GA(1) after transfer is mediated by a light-induced increase in the 2beta-hydroxylation of GA(1) to GA(8). A comparison of the elongation response to GA(1) at early and late stages of de-etiolation provided strong evidence for a change in GA(1) response during de-etiolation, coinciding with the return of GA(1) levels to the normal, homeostatic levels found in light- and dark-grown plants. The emerging picture of the control of shoot elongation by light involves an initial inhibition of elongation by a light-induced decrease in GA(1) levels, with continued inhibition mediated by a light-induced change in the plant's response to the endogenous level of GA(1). Hence the plant uses a change in hormone level to respond to a change in the environment, but over time, homeostasis returns the level of the hormone to normal once the ongoing change in environment is accommodated by a change in the response of the plant to the hormone. PMID- 11027727 TI - Direct evidence for requirement of phosphatidylglycerol in photosystem II of photosynthesis. AB - Phosphatidylglycerol (PG) is considered to play an important role in the ordered assembly and structural maintenance of the photosynthetic apparatus in thylakoid membranes. However, its function in photosynthesis remains poorly understood. In this study we have identified a pgsA gene of Synechocystis sp. PCC6803 that encodes a PG phosphate synthase involved in the biosynthesis of PG. A disruption of the pgsA gene allowed us to manipulate the content of PG in thylakoid membranes and to investigate the function of PG in photosynthesis. The obtained pgsA mutant could grow only in the medium containing PG, and the photosynthetic activity of the pgsA mutant dramatically decreased with a concomitant decrease of PG content in thylakoid membranes when the cells grown in the presence of PG were transferred to the medium without PG. This decrease of photosynthetic activity was attributed to the decrease of photosystem (PS)II activity, but not to the decrease in PSI activity. These findings demonstrate that PG is essential for growth of Synechocystis sp. PCC6803 and provide the first direct evidence that PG plays an important role in PSII. PMID- 11027729 TI - Gibberellin-induced changes in growth anisotropy precede gibberellin-dependent changes in cortical microtubule orientation in developing epidermal cells of barley leaves. Kinematic and cytological studies on a gibberellin-responsive dwarf mutant, M489. AB - We conducted kinematic and cytological studies on "between vein" epidermal cells of the gibberellin (GA)-deficient M489 dwarf mutant of barley (Hordeum vulgare L. Himalaya). GAs affect radial and axial components of cell expansion and cortical microtubule orientation. Adaxial cells in particular expand radially after leaving the elongation zone (EZ), probably as part of leaf unrolling. Exogenous gibberellic acid corrects the mutant's short, wide blades, short EZ, and slow elongation rate. Cell production rates increase more on the adaxial than on the abaxial surface. Cells spend equal periods of time elongating in dwarf and tall plants, but relative elemental growth rates start to decline sooner in the dwarf. GA increased the rate at which longitudinal wall area increased because the increased axial growth more than compensated for reduced radial growth. In dwarf leaves, increased radial expansion was detected in basal parts of the EZ before cortical microtubules lost transverse orientation in the distal elongation zone. We conclude that loss of microtubule orientation is not required for low GA levels to reduce growth anisotropy. PMID- 11027730 TI - Antisense suppression of 2-cysteine peroxiredoxin in Arabidopsis specifically enhances the activities and expression of enzymes associated with ascorbate metabolism but not glutathione metabolism. AB - The aim of this study was to characterize the effect of decreased 2-cysteine peroxiredoxin (2-CP) on the leaf anti-oxidative system in Arabidopsis. At three stages of leaf development, two lines of transgenic Arabidopsis mutants with decreased contents of chloroplast 2-CP were compared with wild type and a control line transformed with an empty vector. Glutathione contents and redox state were similar in all plants, and no changes in transcript levels for enzymes involved in glutathione metabolism were observed. Transcript levels for chloroplastic glutathione peroxidase were much lower than those for 2-CP, and both cytosolic and chloroplastic glutathione peroxidase were not increased in the mutants. In contrast, the foliar ascorbate pool was more oxidized in the mutants, although the difference decreased with plant age. The activities of thylakoid and stromal ascorbate peroxidase and particularly monodehydroascorbate reductase were increased as were transcripts for these enzymes. No change in dehydroascorbate reductase activity was observed, and effects on transcript abundance for glutathione reductase, catalase, and superoxide dismutase were slight or absent. The results demonstrate that 2-CP forms an integral part of the anti-oxidant network of chloroplasts and is functionally interconnected with other defense systems. Suppression of 2-CP leads to increased expression of other anti oxidative genes possibly mediated by increased oxidation state of the leaf ascorbate pool. PMID- 11027731 TI - Differential effects of methyl jasmonate on the expression of the early light inducible proteins and other light-regulated genes in barley. AB - The effects of methyl jasmonate (JA-Me) on early light-inducible protein (ELIP) expression in barley (Hordeum vulgare L. cv Apex) have been studied. Treatment of leaf segments with JA-Me induces the same symptoms as those exhibited by norflurazon bleaching, including a loss of pigments and enhanced light stress that results in increased ELIP expression under both high- and low-light conditions. The expression of both low- and high-molecular-mass ELIP families is considerably down-regulated by JA-Me at the transcript and protein levels. This repression occurs despite increased photoinhibition measurable as a massive degradation of D1 protein and a delayed recovery of photosystem II activity. In JA-Me-treated leaf segments, the decrease of the photochemical efficiency of photosystem II under high light is substantially more pronounced as compared to controls in water. The repression of ELIP expression by JA-Me is superimposed on the effect of the increased light stress that leads to enhanced ELIP expression. The fact that the reduction of ELIP transcript levels is less pronounced than those of light-harvesting complex II and small subunit of Rubisco transcripts indicates that light stress is still affecting gene expression in the presence of JA-Me. The jasmonate-induced protein transcript levels that are induced by JA-Me decline under light stress conditions. PMID- 11027732 TI - Can meristematic activity determine variation in leaf size and elongation rate among four Poa species? A kinematic study. AB - We studied inherent variation in final leaf size among four Poa spp. that live at different elevations. The average final length of leaf 7 of the main stem of the smallest species (Poa alpina) was only one-half that of the largest species (Poa trivialis); it was correlated with leaf elongation rate, but not with the duration of leaf elongation. A faster rate of leaf elongation rate was associated with (a) larger size of the zone of cell expansion, and (b) faster rates of cell production (per cell file) in the meristem, which in turn were due to greater numbers of dividing cells, whereas average cell division rates were very similar for all species (except Poa annua). Also we found that the proliferative fraction equaled 1 throughout the meristem in all species. It was remarkable that rates of cell expansion tended to be somewhat higher in the species with slower growing leaves. We discuss the results by comparing the spatial and material viewpoints, which lead to different interpretations of the role of cell division. Although the presented data do not strictly prove it, they strongly suggest a regulatory role for cell division in determining differences in growth rate among the present four Poa spp. PMID- 11027733 TI - Strategies for the allocation of resources under sulfur limitation in the green alga Dunaliella salina. AB - The effect of sulfur limitation on the partitioning of carbon, nitrogen, and sulfur was investigated in Dunaliella salina. D. salina was able to adapt to 6 microM sulfate; under these conditions, the cells showed reduced growth and photosynthetic rates. Whereas intracellular sulfate was depleted, phosphate, nitrate, and ammonium increased. Amino acids showed a general increase, and alanine became the most abundant amino acid. The activities of four key enzymes of carbon, sulfur, and nitrogen metabolism were differentially regulated: Adenosine 5' triphosphate sulfurylase activity increased 4-fold, nitrate reductase and phosphoenolpyruvate (PEP) carboxylase activities decreased 4- and 11-fold, respectively, whereas carbonic anhydrase activity remained unchanged. Sulfur limitation elicited specific increase or decrease of the abundance of several proteins, such us Rubisco, PEP carboxylase, and a light harvesting complex protein. The accumulation of potentially toxic ammonium indicates an insufficient availability of carbon skeletons. Sulfur deficiency thus induces an imbalance between carbon and nitrogen. The dramatic reduction in PEP carboxylase activity suggests that carbon was diverted away from anaplerosis and possibly channeled into C3 metabolism. These results indicate that it is the coordination of key steps and components of carbon, nitrogen, and sulfur metabolism that allows D. salina to adapt to prolonged sulfur limitation. PMID- 11027734 TI - Gene induction of stilbene biosynthesis in Scots pine in response to ozone treatment, wounding, and fungal infection. AB - The S-adenosyl-L-methionine:pinosylvin-O-methyltransferase (PMT) gene was sequenced from Scots pine (Pinus sylvestris). The open reading frame is arranged in two exons spaced by one 102-bp intron. Promoter regulatory elements such as two "CAAT" boxes and one "TATA" box were identified. Several cis-regulatory elements were recognized: stress-responsive elements (Myb-responsive elements) as well as G, H, and GC boxes. Moreover, elicitor-responsive elements (W boxes) and a sequence resembling the simian virus 40 enhancer core were found. In phloem and needles of control trees, the transcripts of stilbene synthase (STS) and PMT were hardly detectable. Increased ozone fumigation up to 0.3 microL L(-1) enhanced the transcript level of STS and PMT in needles but not in healthy phloem. Wounding, e.g. mock inoculation, of stem-phloem was characterized by a transient increase in STS and PMT transcripts, which was more pronounced in the case of fungal inoculation. Combination of fungal-challenge or mock treatment with ozone resulted in a positive interaction at 0.3 microL L(-1). Scots pine stilbene formation appeared to be induced via STS and PMT gene expression upon ozone and fungal stress as well as wounding. The broad stress-responsiveness is in agreement with the range of various cis-acting elements detected in the STS and PMT promoters. PMID- 11027735 TI - Effects of iron deficiency on the composition of the leaf apoplastic fluid and xylem sap in sugar beet. Implications for iron and carbon transport. AB - The effects of iron deficiency on the composition of the xylem sap and leaf apoplastic fluid have been characterized in sugar beet (Beta vulgaris Monohil hybrid). pH was estimated from direct measurements in apoplastic fluid and xylem sap obtained by centrifugation and by fluorescence of leaves incubated with 5 carboxyfluorescein and fluorescein isothiocyanate-dextran. Iron deficiency caused a slight decrease in the pH of the leaf apoplast (from 6.3 down to 5.9) and xylem sap (from 6.0 down to 5.7) of sugar beet. Major organic acids found in leaf apoplastic fluid and xylem sap were malate and citrate. Total organic acid concentration in control plants was 4.3 mM in apoplastic fluid and 9.4 mM in xylem sap and increased to 12.2 and 50.4 mM, respectively, in iron-deficient plants. Inorganic cation and anion concentrations also changed with iron deficiency both in apoplastic fluid and xylem sap. Iron decreased with iron deficiency from 5.5 to 2.5 microM in apoplastic fluid and xylem sap. Major predicted iron species in both compartments were [FeCitOH](-1) in the controls and [FeCit(2)](-3) in the iron-deficient plants. Data suggest the existence of an influx of organic acids from the roots to the leaves via xylem, probably associated to an anaplerotic carbon dioxide fixation by roots. PMID- 11027736 TI - Responses of sugar beet roots to iron deficiency. Changes in carbon assimilation and oxygen use. AB - Different root parts with or without increased iron-reducing activities have been studied in iron-deficient and iron-sufficient control sugar beet (Beta vulgaris L. Monohil hybrid). The distal root parts of iron-deficient plants, 0 to 5 mm from the root apex, were capable to reduce Fe(III)-chelates and contained concentrations of flavins near 700 microM, two characteristics absent in the 5 to 10 mm sections of iron-deficient plants and the whole root of iron-sufficient plants. Flavin-containing root tips had large pools of carboxylic acids and high activities of enzymes involved in organic acid metabolism. In iron-deficient yellow root tips there was a large increase in carbon fixation associated to an increase in phosphoenolpyruvate carboxylase activity. Part of this carbon was used, through an increase in mitochondrial activity, to increase the capacity to produce reducing power, whereas another part was exported via xylem. Root respiration was increased by iron deficiency. In sugar beet iron-deficient roots flavins would provide a suitable link between the increased capacity to produce reduced nucleotides and the plasma membrane associated ferric chelate reductase enzyme(s). Iron-deficient roots had a large oxygen consumption rate in the presence of cyanide and hydroxisalycilic acid, suggesting that the ferric chelate reductase enzyme is able to reduce oxygen in the absence of Fe(III)-chelates. PMID- 11027737 TI - Hydrogen peroxide yields during the incompatible interaction of tobacco suspension cells inoculated with Phytophthora nicotianae. AB - Rates of H(2)O(2) production by tobacco suspension cells inoculated with zoospores from compatible or incompatible races of the pathogen Phytophthora nicotianae were followed by direct measurement of oxygen evolution from culture supernatants following catalase addition. Rates of HO(2)(*)/O(2)(-) production were compared by following the formation of the formazan of sodium, 3'-[1 [phenylamino-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate. In the incompatible interaction only, both reactive oxygen species (ROS) were produced by the cultured host cells in a minor burst between 0 and 2 h and then in a major burst between 8 and 12 h after inoculation. Absolute levels of H(2)O(2) could not be accurately measured due to its metabolism by host cells, but results are consistent with the majority of H(2)O(2) being formed via dismutation of HO(2)(*)/O(2)(-). The effects of inhibitors of endogenous Cu/Zn superoxide dismutase (diethyldithiocarbamate) and catalase (3-amino-1,2,4 triazole and salicylic acid) were also examined. Yields of ROS in the presence of the inhibitors diphenylene iodonium, allopurinol, and salicylhydroxamic acid suggest that ROS were generated in incompatible host responses by more than one mechanism. PMID- 11027738 TI - Potassium-efflux channels in extensor and flexor cells of the motor organ of Samanea saman are not identical. Effects of cytosolic calcium. AB - Leaflet movements in the mimosa-family tree Samanea saman stem from coordinated volume changes of cells in the leaf motor organs in the adaxial and abaxial motor cells ("flexors" and "extensors"). Shrinking, initiated by dissimilar light signals in extensors and in flexors, depends in both cell types on K(+) efflux via depolarization-dependent potassium (K(D)) channels. To compare between flexor and extensor K(D) channels and to test for a possible interaction of these channels with the Ca(2+)-mobilizing phosphoinositide cascade evoked in these motor cells by the "shrinking signals," we probed the channels with varying (5 nM 3 mM) cytosolic free-Ca(2+) concentration ([Ca(2+)](cyt)) in patch-clamped inside out excised membrane patches. Ca(2+) was not required for K(D) channel activation. [Ca(2+)](cyt) of 600 nM decreased the mean number of open K(D) channels in flexors, as monitored at -30 mV. Detailed analysis revealed that in flexors millimolar [Ca(2+)](cyt) decreased the maximum number of open channels, but simultaneously increased K(D) channel opening probability by negatively shifting the half-maximum-activation voltage by 40 to 50 mV. Thus, the promoting and the inhibitory effects at millimolar [Ca(2+)](cyt) practically cancelled-out. In contrast to flexors, none of the gating parameters of the extensor K(D) channels were affected by [Ca(2+)](cyt). Irrespective of [Ca(2+)](cyt), the steady-state gating of extensor K(D) channels was slightly but significantly more voltage sensitive than that of flexors. The unitary conductances of flexor and extensor K(D) channels were similar and decreased by approximately 20% at millimolar [Ca(2+)](cyt). It is intriguing that the extensor K(D) channels were significantly less K(+) selective than those in flexors. PMID- 11027739 TI - Long-term effects of budesonide or nedocromil in children with asthma. AB - BACKGROUND: Antiinflammatory therapies, such as inhaled corticosteroids or nedocromil, are recommended for children with asthma, although there is limited information on their long-term use. METHODS: We randomly assigned 1041 children from 5 through 12 years of age with mild-to-moderate asthma to receive 200 microg of budesonide (311 children), 8 mg of nedocromil (312 children), or placebo (418 children) twice daily. We treated the participants for four to six years. All children used albuterol for asthma symptoms. RESULTS: There was no significant difference between either treatment and placebo in the primary outcome, the degree of change in the forced expiratory volume in one second (FEV1, expressed as a percentage of the predicted value) after the administration of a bronchodilator. As compared with the children assigned to placebo, the children assigned to receive budesonide had a significantly smaller decline in the ratio of FEV1 to forced vital capacity (FVC, expressed as a percentage) before the administration of a bronchodilator (decline in FEV1:FVC, 0.2 percent vs. 1.8 percent). The children given budesonide also had lower airway responsiveness to methacholine, fewer hospitalizations (2.5 vs. 4.4 per 100 person-years), fewer urgent visits to a caregiver (12 vs. 22 per 100 person-years), greater reduction in the need for albuterol for symptoms, fewer courses of prednisone, and a smaller percentage of days on which additional asthma medications were needed. As compared with placebo, nedocromil significantly reduced urgent care visits (16 vs. 22 per 100 person-years) and courses of prednisone. The mean increase in height in the budesonide group was 1.1 cm less than in the placebo group (22.7 vs. 23.8 cm, P=0.005); this difference was evident mostly within the first year. The height increase was similar in the nedocromil and placebo groups. CONCLUSIONS: In children with mild-to-moderate asthma, neither budesonide nor nedocromil is better than placebo in terms of lung function, but inhaled budesonide improves airway responsiveness and provides better control of asthma than placebo or nedocromil. The side effects of budesonide are limited to a small, transient reduction in growth velocity. PMID- 11027740 TI - Effect of long-term treatment with inhaled budesonide on adult height in children with asthma. AB - BACKGROUND: Short-term studies have shown that inhaled corticosteroids may reduce the growth of children with asthma. However, the effect of long-term treatment on adult height is uncertain. METHODS: We conducted a prospective study in children with asthma to examine the effect of long-term treatment with inhaled budesonide on adult height. We report on 211 children who have attained adult height: 142 budesonide-treated children with asthma, 18 control patients with asthma who have never received inhaled corticosteroids, and 51 healthy siblings of patients in the budesonide group, who also served as controls. RESULTS: The children in the budesonide group attained adult height after a mean of 9.2 years of budesonide treatment (range, 3 to 13) at a mean daily dose of 412 microg (range, 110 to 877). The mean cumulative dose of budesonide was 1.35 g (range, 0.41 to 3.99). The mean differences between the measured and target adult heights were +0.3 cm (95 percent confidence interval, -0.6 to + 1.2) for the budesonide-treated children, -0.2 cm (95 percent confidence interval, -2.4 to +2.1) for the control children with asthma, and +0.9 cm (95 percent confidence interval, -0.4 to +2.2) for the healthy siblings. The adult height depended significantly (P<0.001) on the child's height before budesonide treatment. Although growth rates were significantly reduced during the first years of budesonide treatment, these changes in growth rate were not significantly associated with adult height. CONCLUSIONS: Children with asthma who have received long-term treatment with budesonide attain normal adult height. PMID- 11027741 TI - Entrainment of free-running circadian rhythms by melatonin in blind people. AB - BACKGROUND: Most totally blind people have circadian rhythms that are "free running" (i.e., that are not synchronized to environmental time cues and that oscillate on a cycle slightly longer than 24 hours). This condition causes recurrent insomnia and daytime sleepiness when the rhythms drift out of phase with the normal 24-hour cycle. We investigated whether a daily dose of melatonin could entrain their circadian rhythms to a normal 24-hour cycle. METHODS: We performed a crossover study involving seven totally blind subjects who had free running circadian rhythms. The subjects were given 10 mg of melatonin or placebo daily, one hour before their preferred bedtime, for three to nine weeks. They were then given the other treatment. The timing of the production of endogenous melatonin was measured as a marker of the circadian time (phase), and sleep was monitored by polysomnography. RESULTS: At base line, the subjects had free running circadian rhythms with distinct and predictable cycles averaging 24.5 hours (range, 24.2 to 24.9). These rhythms were unaffected by the administration of placebo. In six of the seven subjects the rhythm was entrained to a 24.0-hour cycle during melatonin treatment (P<0.001). After entrainment, the subjects spent less time awake after the initial onset of sleep (P=0.05) and the efficiency of sleep was higher (P=0.06). Three subjects subsequently participated in a trial in which a 10-mg dose of melatonin was given daily until entrainment was achieved. The dose was then reduced to 0.5 mg per day over a period of three months; the entrainment persisted, even at the lowest dose. CONCLUSIONS: Administration of melatonin can entrain circadian rhythms in most blind people who have free running rhythms. PMID- 11027742 TI - Twelve years' experience with national sharing of HLA-matched cadaveric kidneys for transplantation. AB - BACKGROUND: In October 1987, the United Network for Organ Sharing (UNOS) established a national kidney-sharing program to increase the number of HLA matched transplantations. Since then, over 7500 cadaveric kidneys have been shipped to centers in 48 states for transplantation to HLA-matched patients. We evaluated the efficacy of the program during its first 12 years of operation. METHODS: We compared the rates of rejection and actuarial graft survival for 7614 HLA-matched and 81,364 HLA-mismatched cadaveric kidney transplantations reported to the UNOS Scientific Registry between October 1987 and September 1999. To assess the effects of the extended period of ischemia associated with shipping HLA-matched kidneys, we identified 3562 pairs of cadaveric kidneys in which one kidney went to an HLA-matched recipient and the other went to an HLA-mismatched recipient. RESULTS: The estimated 10-year rate of graft survival was 52 percent for HLA-matched transplants, as compared with 37 percent for HLA-mismatched transplants. The estimated half-lives of the transplants were 12.5 years and 8.6 years, respectively, and the mean duration of cold ischemia was 23 hours and 22 hours, respectively. After adjustment for the effects of demographic characteristics, at 10 years the overall rates of graft survival and the rates of functional-graft survival (with data censored on patients who died with a functioning graft) were 10 percent and 11 percent higher, respectively, for HLA matched transplants than for HLA-mismatched transplants. Among 3562 pairs of kidneys, HLA-matched transplants had higher rates of survival, a lower incidence of episodes of rejection, and a lower risk of loss as a result of rejection. CONCLUSIONS: A superior graft outcome with little increase in the duration of cold ischemia justifies national sharing of HLA-matched kidney transplants. PMID- 11027743 TI - Images in clinical medicine. Schistosoma haematobium. PMID- 11027744 TI - Primary care for survivors of breast cancer. PMID- 11027746 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 31-2000. A 32-year-old man with a lesion of the urinary bladder. PMID- 11027745 TI - Genetic defects of intracellular-membrane transport. PMID- 11027747 TI - Asthma, steroids, and growth. PMID- 11027748 TI - Melatonin, circadian rhythms, and sleep. PMID- 11027749 TI - Correction: Pyomyositis Complicating Acute Bacterial Endocarditis in an Intravenous Drug User. PMID- 11027750 TI - Correction: The Immune System (First of Two Parts). PMID- 11027751 TI - Traumatic glenohumeral bone defects and their relationship to failure of arthroscopic Bankart repairs: significance of the inverted-pear glenoid and the humeral engaging Hill-Sachs lesion. AB - PURPOSE: Our goal was to analyze the results of 194 consecutive arthroscopic Bankart repairs (performed by 2 surgeons with an identical suture anchor technique) in order to identify specific factors related to recurrence of instability. TYPE OF STUDY: Case series. MATERIALS AND METHODS: We analyzed 194 consecutive arthroscopic Bankart repairs by suture anchor technique performed for traumatic anterior-inferior instability. The average follow-up was 27 months (range, 14 to 79 months). There were 101 contact athletes (96 South African rugby players and 5 American football players). We identified significant bone defects on either the humerus or the glenoid as (1) "inverted-pear" glenoid, in which the normally pear-shaped glenoid had lost enough anterior-inferior bone to assume the shape of an inverted pear; or (2) "engaging" Hill-Sachs lesion of the humerus, in which the orientation of the Hill-Sachs lesion was such that it engaged the anterior glenoid with the shoulder in abduction and external rotation. RESULTS: There were 21 recurrent dislocations and subluxations (14 dislocations, 7 subluxations). Of those 21 shoulders with recurrent instability, 14 had significant bone defects (3 engaging Hill-Sachs and 11 inverted-pear Bankart lesions). For the group of patients without significant bone defects (173 shoulders), there were 7 recurrences (4% recurrence rate). For the group with significant bone defects (21 patients), there were 14 recurrences (67% recurrence rate). For contact athletes without significant bone defects, there was a 6.5% recurrence rate, whereas for contact athletes with significant bone defects, there was an 89% recurrence rate. CONCLUSIONS: (1) Arthroscopic Bankart repairs give results equal to open Bankart repairs if there are no significant structural bone deficits (engaging Hill-Sachs or inverted-pear Bankart lesions). (2) Patients with significant bone deficits as defined in this study are not candidates for arthroscopic Bankart repair. (3) Contact athletes without structural bone deficits may be treated by arthroscopic Bankart repair. However, contact athletes with bone deficiency require open surgery aimed at their specific anatomic deficiencies. (4) For patients with significant glenoid bone loss, the surgeon should consider reconstruction by means of the Latarjet procedure, using a large coracoid bone graft. PMID- 11027752 TI - Arthroscopic treatment of symptomatic shoulders with minimally displaced greater tuberosity fracture. AB - PURPOSE: The purpose of this study is to describe arthroscopic findings and the results of arthroscopic treatment of patients with chronic shoulder pain caused by a minimally displaced fracture of the greater tuberosity. TYPE OF STUDY: This is a retrospective case series in a consecutive sample of 23 patients with a minimally displaced or nondisplaced fracture of the greater tuberosity who underwent outcome analysis after arthroscopic treatment. MATERIALS AND METHODS: Twenty-three patients with chronic shoulder pain beyond 6 months after fracture of the greater tuberosity underwent arthroscopic treatment and were retrospectively assessed after an average of 29 months (range, 22 to 40 months). There were 18 men and 5 women with the average age of 39 years (range, 24 to 61 years). Fourteen fractures were isolated and 9 were related to acute anterior instability episode. The average displacement of the fracture was 2.3 mm (range, 0 to 4 mm) as shown on anteroposterior view plain radiographs. RESULTS: At the time of arthroscopy, all patients had partial-thickness rotator cuff tears on the articular surface. The cuff tears were located on the tuberosity fracture area and were an Ellman grade I to II in depth. After arthroscopic debridement or repair of the tear, depending on the condition of the tear itself, as well as subacromial decompression, the UCLA score results were good to excellent in 20 and fair in 3 patients. Nineteen of the patients had returned to the previous level of activities. However, the patients engaged in overhead sports activity had a lower level of return to activity (P =.034). CONCLUSIONS: A partial thickness rotator cuff tear on the articular surface should be considered in patients with chronic shoulder pain after a minimally displaced fracture of the greater tuberosity. Arthroscopic debridement or repair is an appropriate procedure. PMID- 11027753 TI - Treatment of osteoarthritis of the elbow: a comparison of open and arthroscopic debridement. AB - PURPOSE: To assess the effectiveness of open and minimally invasive techniques in the debridement of osteoarthritis of the elbow, we compared the Outerbridge Kashiwagi (O-K) procedure with an arthroscopic modification in which arthroscopic debridement and fenestration of the olecranon fossa was performed. TYPE OF STUDY: The study took the form of a nonrandomized control trial in which subjects were allocated to a treatment depending on the hospital of presentation. MATERIALS AND METHODS: Assessment using the Mayo Clinic elbow function chart enabled comparison of the outcome in 18 cases treated by the O-K procedure and 26 patients treated by arthroscopic debridement and fenestration of the olecranon fossa. Mean follow up was 35.3 months (minimum 12 months). Of the patients treated by the O-K procedure, 14 were men and 4 were women with a mean age of 55 years. In 83% of patients, the diagnosis was primary osteoarthritis, with the remainder post traumatic arthritis. The patients treated by arthroscopic debridement and fenestration of the olecranon fossa included 24 men and 2 women with a mean age of 46 years, and a diagnosis of primary osteoarthritis in 91% and post-traumatic arthritis in the remainder. No patients were excluded from the study or refused to be included. RESULTS: Both procedures were shown to be effective, with no major complications. Patients treated by arthroscopic debridement and fenestration of the olecranon fossa achieved better relief of pain (P <.10), whereas those patients undergoing the O-K procedure achieved significantly greater improvement in range of flexion (P <.05). No difference between the procedures in terms of patient-perceived overall effectiveness of the surgery was found. CONCLUSIONS: In conclusion, in the treatment of osteoarthritis of the elbow, arthroscopic debridement and fenestration of the olecranon fossa may be a more suitable procedure when painful symptoms predominate. In contrast, the O-K procedure is a significantly better procedure for improving the range of flexion where this is a particular problem. PMID- 11027755 TI - Practical considerations in anterior cruciate ligament replacement surgery. AB - The endoscopic method of anterior cruciate ligament (ACL) reconstruction using a patellar tendon graft that is secured with interference screws is a commonly performed procedure. It has many potential pitfalls, the majority of which are secondary to technical errors. Prevention of errors starts with a full knowledge of normal ACL anatomy and any discrepancies with the size and shape of the graft substitute. Accurate tibial and femoral tunnel placement is essential and involves using consistent intra-articular landmarks and achieving specific radiographic criteria. Accurate tunnel placement minimizes graft excursion and impingement against the roof of the intercondylar notch. This will result in maximum knee stability and motion. Much has been written about the principles of graft-tunnel mismatch and interference screw-bone plug divergence. The clinical implications of these potential sources of error remain somewhat controversial and are discussed in this article. PMID- 11027754 TI - Tunnel expansion after hamstring anterior cruciate ligament reconstruction with 1 incision EndoButton femoral fixation. AB - In this study, we compared a study group of 20 patients who underwent anterior cruciate ligament (ACL) reconstruction with hamstring autografts using a 1 incision endoscopic technique verses a control group of 20 patients using a 2 incision technique. The patient groups were compared based on increase of bone tunnel diameter seen radiographically, physical examination, and arthrometer measurements. The 1-incision technique differed from the 2-incision technique in 2 ways: an EndoButton femoral fixation system and drilling of the femoral tunnel through the tibial tunnel (transtibial). This study shows that the majority of tunnel diameter measurements for the 1-incision ACL reconstruction technique were greater than those of the 2-incision ACL reconstruction technique using autologous hamstring tendons, at both 3 and 12 months of follow-up. Between 3 and 12 months follow-up, there was no statistical differences in tunnel enlargement between the 2 groups of patients. The measured tunnel enlargement, therefore, would have occurred before the 3-month follow-up. There was no significant difference in the Lachman or arthrometer testing in either group of patients at the termination of this study. This indicates that, although tunnel expansion is significant, the increased expansion is not related to increased knee laxity in the first year. PMID- 11027756 TI - The treatment of complete posterior cruciate ligament tears. PMID- 11027757 TI - Anterior and posterior cruciate ligament rupture after thermal treatment. AB - SUMMARY: This article reports the first published case of spontaneous, simultaneous rupture of both the anterior and posterior cruciate ligaments 3 months after radiofrequency thermal probe treatment. The patient had no previous traumatic injury to the knee. Simultaneous rupture occurred under minimal physiologic load. Clinical, radiologic, and surgical findings are presented. PMID- 11027758 TI - Distal femoral fracture through the screw hole of a ligament augmentation device fixation. AB - SUMMARY: Complications associated with fixation of artificial ligaments in augmented repair of the anterior cruciate ligament (ACL) have been reported throughout the literature. However, fractures following ligament augmentation device (LAD) fixation appear to be rare. We report the case of a 43-year-old woman, injured in a road accident, who sustained a depressed fracture of the tibial plateau and knee instability. The fracture was reduced and the medial collateral ligament and the menisci were sutured. The torn ACL was repaired using the Marshall technique and augmented with an LAD in an over-the-top technique. Twenty-five months postoperatively, the patient sustained a distal femoral fracture through the screw hole of the former LAD fixation after a simple fall on the street. PMID- 11027759 TI - Articular arc length mismatch as a cause of failed bankart repair. AB - The purpose of this article is to recognize an often overlooked cause of failed anterior stabilization procedures of the shoulder. Hill-Sachs lesions are common occurrences after anterior dislocation. But despite the frequency of occurrence, they are commonly not addressed in the treatment of recurrent anterior dislocation. The Hill-Sachs lesion produces a shortened rotational arc length of the humeral head on the glenoid; any engagement of the defect with the glenoid produces a sensation in the patient of subluxation/dislocation. However, instead of subluxation/dislocation, the sensation may be caused by mismatch between the articular arc length of the humeral head and the glenoid. The literature concerning the Hill-Sachs lesion and its association with recurrent dislocation has been reviewed. There is no recognition in the literature of this articular arc length mismatch as a cause of failed anterior stabilization procedures. We present a case in which a Hill-Sachs lesion was the cause of a perceived recurrent subluxation/dislocation of the shoulder. PMID- 11027760 TI - Detached deltoid during arthroscopic subacromial decompression. AB - SUMMARY: Arthroscopic subacromial decompression has become increasingly popular as an operative treatment for shoulder impingement syndrome. It is generally a safe procedure with low morbidity and very few complications. A case where the deltoid became detached during arthroscopic subacromial decompression is presented as an example of the perils of overaggressive subacromial decompression. This is the first reported case of such a complication related to arthroscopic subacromial decompression. PMID- 11027761 TI - Chondral injury following meniscal repair with a biodegradable implant. AB - This case report presents chondral damage associated with a resorbable meniscal repair implant. Although the devices may not have been inserted properly, surgeons using these implants should be aware of the potential severe chondral injury that may occur. PMID- 11027762 TI - Chondral injury after meniscal repair with bioabsorbable arrows. AB - SUMMARY: Meniscal repair is a frequently performed procedure. Repair may be accomplished using several different techniques including inside-out, outside-in, and all-inside instrumentation. The Meniscus Arrow (Bionx, Blue Bell, PA) is an all-inside technique that has gained considerable popularity in the past 3 years. We report and illustrate a potential complication with this implant. Arrow fracture with condyle grooving should be considered in patients who have undergone repair with the Meniscus Arrow and complain of late postoperative symptoms. PMID- 11027763 TI - Nonunited avulsion fractures of the intercondylar eminence of the tibia. AB - We report 3 cases of nonunited avulsion fracture of the intercondylar eminence of the tibia. Characteristics of the clinical symptoms were limitation of knee extension, pain at knee extension, and a small amount of anterior instability. For the patient with a normal anterior cruciate ligament (ACL), the fragment was fixed by sutures with a button after curettage of the fibrous tissue. Two patients underwent endoscopic ACL reconstruction using quadrupled semitendinosus tendon because their ACLs had degenerated. Since the technique of ACL reconstruction under arthroscopic control has been well refined, endoscopic ACL reconstruction is considered to be 1 of the surgical options for nonunited avulsion fracture with symptomatic anterior instability if the ACL is degenerated. PMID- 11027764 TI - Saphenous neuralgia after arthroscopically assisted anterior cruciate ligament reconstruction with a semitendinosus and gracilis tendon graft. AB - A case report of saphenous neuralgia following arthroscopically assisted anterior cruciate ligament reconstruction with hamstring tendons is presented. The patient complained of paresthesia in the anteromedial region of the lower leg and tenderness at the medial side of the knee without motor or reflex abnormalities. Because saphenous neuralgia can mimic disorders of the knee, peripheral vascular disease, and lumbar nerve root compression, diagnosis can be confirmed by anesthetic blockade. The patient underwent saphenous neurolysis. Six months after surgery, the patient had normal cutaneous sensation at the medial aspect of the lower leg and ankle and she no longer complained of any painful dysesthesia. To minimize the risk of damaging the saphenous nerve when harvesting hamstring tendons, the knee should be flexed and the hip external rotated. PMID- 11027765 TI - Localized pigmented villonodular synovitis presenting as recurrent dislocation of the patella. AB - A 31-year-old woman presented with a 1-year history of recurrent dislocation of the patella. On examination, she was found to have a mass at the medial patellofemoral joint line. Arthroscopy showed this to be a vascular synovial tumor extending into the medial patellofemoral joint and causing lateral deviation of the patella. The mass was resected with resolution of her symptoms and histologic examination showed this to be pigmented villonodular synovitis. There are an increasing number of case reports of these lesions causing varying mechanical symptoms in the knee and they should be considered as a differential diagnosis. PMID- 11027766 TI - Avoiding the labrum in hip arthroscopy. AB - SUMMARY: Iatrogenic joint damage is a potential complication of any arthroscopic procedure. The anatomic constraints of the hip cause particular concern. The greatest risks are perforation of the acetabular labrum and scuffing of the articular surface. Careful attention to the details of the technique described can reduce the likelihood of this problem. With current technology, some occasional damage is unavoidable. However, all steps should be taken to minimize the likelihood and magnitude of such occurrences. PMID- 11027767 TI - Posterior trans-septal portal for arthroscopic surgery of the knee joint. AB - The authors introduce a safe arthroscopic technique of making a portal through the posterior septum of the knee joint, posterior to the posterior cruciate ligament (PCL). This posterior trans-septal portal makes it possible for the arthroscope or the working instruments inserted through the posteromedial portal to reach the posterolateral compartment. With this portal, complete arthroscopic visualization of the posterior compartment and easier arthroscopic procedures for the posterior compartment of the knee joints are possible, including the posterior and posterosuperior aspect of the both femoral condyles, the posterior horns of both menisci, the posterior and inferior portion of the PCL, the posterior meniscofemoral ligament, the posterior septum, and the posterior capsule. We have used this technique in more than 150 knees for arthroscopic total synovectomy for arthritis of various origins, arthroscopic PCL reconstructions, arthroscopic removal of encapsulated loose bodies or tumors located behind the PCL, arthroscopic repairs of tears in the posterior horns of the medial menisci using the all-inside technique, arthroscopic pullout suture fixation of the avulsion fracture of tibial attachment of PCL, and others. No complications such as injuries of the popliteal neurovascular structures have occurred in our experience. PMID- 11027768 TI - Arthroscopic double-bundled posterior cruciate ligament reconstruction with quadriceps tendon-patellar bone autograft. AB - An arthroscopic technique for double-bundled reconstruction for posterior cruciate ligament with quadriceps tendon-patellar bone autograft is presented. Anterolateral and posteromedial tunnels were created to simulate and reproduce the double-bundle structure of the posterior cruciate ligament. The bone plug is situated at the tibial tunnel and fixed by a titanium interference screw. Each of the bundles of tendon graft is rigidly fixed at the femoral tunnel with a bioabsorbable screw. PMID- 11027769 TI - Giving back PMID- 11027770 TI - Arthroscopic arthrodesis of the ankle joint. PMID- 11027772 TI - Massive intraosseous ganglion of the talus: reconstruction of the articular surface of the ankle joint. AB - We report on the outcome after autologous chondrocyte and spongiosal bone transplantation in a case of a massive intraosseous ganglion of the talus in a young patient. A 24-year-old man suffered from decreased ankle joint motion, recurrent swelling, and pain. Diagnostic evaluation by plain radiographs, computed tomography, and magnetic resonance imaging revealed cystic lesions in the head and the body of the talus with additional involvement of the cartilage surface. Operative treatment consisted primarily of an initial diagnostic arthroscopy, which established grade VI articular damage according to the arthroscopic classification of Bauer and Jackson. Pathological examination of intralesional biopsy tissue revealed the existence of an intraosseous ganglion. Additionally, healthy cartilage biopsy specimens were obtained and sent for chondrocyte extraction and cultivation with 60 mL of autologous serum. To retain the function of the ankle joint and to minimize the number of necessary operative interventions, 3 weeks after the initial arthroscopic operation, we performed a simultaneous curettage of the cystic lesion followed by autologous spongiosal bone and cultivated chondrocytes transplantation of the talus. Continuous passive motion was applied postoperatively and full weight bearing was allowed after 8 weeks. There were no complications. The clinical result after 18 months was excellent, with a fully functional, pain-free, and weight-bearing ankle joint. The postoperative evaluation score of Finsen (modified Weber score) of 2/6 = 0.3 showed an improvement comparison with the preoperative value of of 21/6 = 3.5 (0 = normal, 4 = pathologic). We encountered no complications postoperatively. Clinical success was achieved by this method of treatment on a patient too young to be treated through arthrodesis. PMID- 11027773 TI - Localized pigmented villonodular synovitis as a rare cause of chronic anterolateral ankle pain in an equestrienne. AB - An unusual case of localized pigmented villonodular synovitis of the ankle as a rare cause of chronic anterolateral ankle pain in a 16-year-old horsewoman is presented. Intra-articular nodular forms of pigmented villonodular synovitis can only be diagnosed arthroscopically, macroscopic and microscopic aspects being typical. We believe that this lesion is more likely a reactive process secondary to repetitive microtrauma rather than a true neoplasm. Our patient presented with pathology in the left ankle, the side by which one mounts and dismounts a horse, forcing, in both activities, ankle dorsiflexion. Moreover, an English saddle was used by our patient, upon which one rides with the ankle maintained in dorsiflexion. At arthroscopy, the soft-tissue mass was seen to be entrapped in the joint between the talus and the tibia at dorsiflexion of the ankle. This had caused a slowly progressive enlargement of the lesion because of fibrosis resulting from reactive inflammation associated with this repetitive microtrauma, thus causing irritation, pain, and synovitis due to impingement. PMID- 11027774 TI - Loose bodies after arthroscopic osteochondral autograft in osteochondritis dissecans of the knee. AB - We report a case of loose bodies from the donor site as a complication after the osteochondral autograft for the treatment of osteochondritis dissecans. Eight months after surgery, 3 osteochondral loose fragments, having dislodged from the donor sites of the osteochondral autograft, were found in the posteromedial portion, posterolateral portion, and anterior compartment of the knee, respectively. A large osteochondral defect can be treated successfully with arthroscopic autogenous bone graft. When filling the donor site with the recipient bone core, only the cancellous portion should be inserted into the donor socket. PMID- 11027775 TI - Chondral lesions after arthroscopic meniscus repair using meniscus arrows. AB - Meniscus repair using bioabsorbable devices has become popular in the last few years. Good clinical results have been reported and few complications have been published. This report describes the case of a 37-year-old male patient with a lateral meniscus repair using 4 Meniscus Arrows (Bionx Implants, Blue Bell, PA). Postoperatively, repeated episodes of intra-articular effusions have occurred. A second-look arthroscopy 8 months after the reconstruction showed that the meniscus tear had not healed and revealed the presence of chondral damage corresponding to the location of the arrows in the posterior area of the lateral femoral condyle. Surgeons using the Meniscus Arrow should be aware of this possible postoperative complication. PMID- 11027776 TI - Technical note: rotator cuff tear. Intraoperative electrical stimulation of the suprascapular nerve. AB - We describe a method of placing an electrically conductive catheter onto the suprascapular nerve for intraoperative electrical stimulation of the nerve. This causes contractions of the supraspinatus and infraspinatus muscles that indicate rotator cuff viability, suitability for repair, direction in which the fibers contract, and the tear pattern. PMID- 11027777 TI - Toxoplasma gondii secretes a calcium-independent phospholipase A(2). AB - Phospholipases A(2) (PLA(2)) play an important role in Toxoplasma gondii host cell penetration. They are also key enzymes in the host cell response to the parasite invasion. PLA(2) hydrolyse cellular phospholipids, releasing multiple inflammatory lipidic mediators. We have investigated the biochemical characterisation of T. gondii PLA(2) activity in a mouse-cultured tachyzoite homogenate and in the peritoneal exudate from infected mice, using the hydrolysis of a fluorescent phosphatidylglycerol labelled at the sn-2 position. Spectrofluorimetry and thin-layer chromatography showed a PLA(2) activity (about 0.5-2 nmol/min per mg), calcium-independent, secreted into infected mice peritoneal exudate, with a broad pH activity ranging between 6.5 and 9.5 and resistant to a great number of potential PLA(2) inhibitors except dithio nitrobenzoic acid (1 mM). An associated phospholipase A(1) activity was also displayed. These results suggest that Toxoplasma gondii displays specific phospholipases different from host enzymes and probably involved at critical steps of infectious cycle. PMID- 11027778 TI - Species associations among larval helminths in an amphipod intermediate host. AB - Larval helminths that share the same intermediate host may or may not also share the same definitive hosts. If one or more of these helminth species can manipulate the phenotype of the intermediate host, there can be great advantages or severe costs for other helminths resulting from co-occurring with a manipulator, depending on whether they have the same definitive host or not. Among 2372 specimens of the amphipod Echinogammarus stammeri collected from the river Brenta, northern Italy, there was a positive association between two acanthocephalan species with the same fish definitive hosts, the relatively common Pomphorhynchus laevis and the much less prevalent Acanthocephalus clavula. The number of cystacanths of P. laevis per infected amphipod, which ranged from one to five, did not influence the likelihood that the amphipod would also host A. clavula. A third acanthocephalan species, Polymorphus minutus,which matures in birds, showed no association with either of the two other species. These results show that associations among helminth species in intermediate hosts are not random, and are instead the product of selection favouring certain pathways of transmission. PMID- 11027779 TI - Nestedness, anti-nestedness, and the relationship between prevalence and intensity in ectoparasite assemblages of marine fish: a spatial model of species coexistence. AB - Nested species subset patterns consist in a hierarchical structure of species composition in related assemblages, with the species found in depauperate assemblages representing non-random subsets of progressively richer ones. This pattern has been found at the infracommunity level in about a third of the fish ectoparasite assemblages studied to date. Here we present evidence for another non-random structural pattern in assemblages of fish ectoparasites, anti nestedness, which corresponds to situations in which parasite species are always absent from infracommunities richer than the most depauperate one in which they occur. We show that this pattern is exactly as common as nestedness, and that anti-nested assemblages are characterised by significantly lower prevalence and mean intensities of parasites than nested assemblages. In addition, we found a positive relationship between the prevalence and the mean intensity of parasites across the different assemblages. We propose a link between the nestedness/anti nestedness continuum and the prevalence-intensity relationship that may involve colonisation-extinction processes. The results presented here suggest that, although nestedness may not be common in parasite communities, other departures from random species assembly are possible, and that some form of structure may be present in many communities. The continuum between nestedness and anti-nestedness also has implications for recent models of species coexistence in communities. PMID- 11027780 TI - Nestedness in assemblages of gyrodactylids (Monogenea: Gyrodactylidea) parasitising two species of cyprinid--with reference to generalists and specialists. AB - The structure of gyrodactylid assemblages in individual fishes of two species of cyprinid was determined. A total of 100 specimens of minnow, Phoxinus phoxinus, and 137 specimens of roach, Rutilus rutilus, were investigated for presence of gyrodactylids. Host specificity, specialists vs. generalists, was noted in each host fish. A nested pattern was recorded in parasite assemblages of minnow, the host with a dominant number of specialist gyrodactylids. A non-nested pattern was observed in parasite assemblages of roach, the host with a dominant number of generalist gyrodactylids. The host specificity appears to be a meaningful factor that determines the pattern of gyrodactylid assemblages of both fish hosts. PMID- 11027781 TI - Genetic variation within the ticks Ixodes holocyclus and Ixodes cornuatus from south-eastern Australia. AB - Ticks from mainland Australia (Victoria, New South Wales and Queensland) and Tasmania, identified morphologically as either Ixodes holocyclus or Ixodes cornuatus, were compared genetically using 24 enzyme loci. The results showed that ticks from three localities in Victoria were genetically similar to I. cornuatus in Tasmania, but both groups had fixed genetic differences at >45% of loci compared with other ticks on the mainland. In addition, there were fixed genetic differences at 0-60% of loci among I. holocyclus from different localities on the mainland. Ixodes holocyclus samples could be divided into four distinct clusters (with fixed genetic differences >15%), three of which were represented by one or two specimens. Nonetheless, these electrophoretic data suggest that I. holocyclus represents a species complex. The results also showed that the morphological criteria used to identify specimens were not always accurate because several specimens had been mis-identified morphologically. Despite limitations with the morphological identification, this study has demonstrated that I. cornuatus can be distinguished from the I. holocyclus species complex using six enzyme loci, providing the foundation for a re examination of morphological characteristics. The present study has shown that I. cornuatus and the I. holocyclus complexes have a greater distribution than previously reported, with both occurring in sympatry at Cape Patterson, on the southern coastline of Victoria. PMID- 11027782 TI - Immunization of dogs with a canine herpesvirus vector expressing Neospora caninum surface protein, NcSRS2. AB - In order to develop a vaccine against Neospora caninum in dogs, we constructed recombinant canine herpesvirus (CHV) expressing N. caninum surface protein, NcSRS2. Indirect immunofluorescence indicated that the antigenic structure of the recombinant NcSRS2 was similar to the authentic parasite protein. The dogs immunised with recombinant virus produced IgG antibody to N. caninum, and their sera recognised the parasite protein on Western blot. The dogs inoculated with recombinant virus showed no clinical symptoms and infectious CHV was not recovered from the dogs, suggesting that recombinant CHV expressing N. caninum proteins may lead to a vaccine against neosporosis in dogs. PMID- 11027783 TI - Vaccination of young lambs against infection with Nematodirus battus using gamma irradiated larvae. AB - Helminthologically naIve 6-week-old Suffolk lambs were given 1-3 doses of 20000 gamma-irradiated infective larvae (L3) of the nematode Nematodirus battus at weekly intervals. Following an anthelmintic drench they were challenged with 50000 viable L3 at 10 weeks of age. Nematode worm burdens 14 days post-challenge showed a significant (P<0.01) 66% reduction in the single vaccine dose group. The two and three dose groups had mean worm burdens which were 30 and 42% lower than controls, respectively, although these were not statistically significant. There was little measurable stimulation of the immune system in the vaccinated lambs, suggesting that the repeatedly dosed animals may have developed immunological unresponsiveness to the parasite. PMID- 11027784 TI - Sensitivity of the nested-polymerase chain reaction (PCR) assay for Brugia malayi and significance of 'free' DNA in PCR-based assays. AB - The blood filtration method was used as the gold standard to determine the detection level of simple blood-spot sampling and nested-polymerase chain reaction (PCR) for Brugia malayi. Of 100 samples, 48 were filtration-positive. Of these, 26 had microfilaria counts that were low enough (<1-29 microfilariae/ml) to accurately assess the limit of detection by nested-PCR. Nested-PCR consistently detected B. malayi DNA in samples with > or = 10 microfilariae/ml. Post-filtration, microfilaria-depleted, blood-spots from microfilaria-positive samples were screened by nested-PCR and B. malayi specific 'free' DNA was detected in 51.7% of these samples. There was no evidence for 'free' DNA in microfilaria-negative individuals from this endemic community. PMID- 11027785 TI - The beta-tubulin gene of Babesia and Theileria parasites is an informative marker for species discrimination. AB - A fragment of the beta-tubulin gene was polymerase chain reaction (PCR) amplified from genomic DNAs of Babesia bovis, Babesia bigemina, Babesia divergens, Babesia major, Babesia caballi, Babesia equi, Babesia microti, Theileria annulata and Theileria sergenti. Single amplification products were obtained for each of these species, but the size of the amplicons varied from 310 to 460 bp. Sequence analysis revealed that this variation is due to the presence of a single intron, which ranged from 20 to 170 bp. The extensive genetic variability at the beta tubulin locus has been exploited to develop two types of species identification assays. The first assay can be used on samples containing mostly parasite DNA, like those prepared from infected erythrocytes. Following PCR amplification, the species identification is obtained directly from the size of the products (for Babesia species infecting human or horse) or using a simple PCR-restriction fragment length polymorphism (RFLP) protocol (for Babesia species infecting cattle). The second assay can be used on samples prepared from whole blood, that contain both parasite and host DNAs. In this case, due to the strong conservation of the beta-tubulin gene, co-amplification of a gene fragment from the host DNA was observed. A nested PCR assay was developed for the specific amplification of parasite DNA, using a primer designed to span the exon-intron boundary. Direct identification of Babesia species infecting human and horse is again obtained after the electrophoretic separation of the amplification products, while for Babesia and Theileria species infecting cattle, differentiation is based on a nested PCR-RFLP protocol. These methods may be used for the simultaneous identification of horses and cattle carrying multiple parasites by means of a single PCR or using the PCR-RFLP protocol. PMID- 11027786 TI - Strain and stage specific variation in Toxoplasma gondii antigens. AB - The antigenic profile of virulent (RH, ENT, Martin) and avirulent (RRA, DEG, ME49) Toxoplasma strains was compared directly by western blotting using a panel of immune mouse sera. Dominant antigens of approximate MR 30-33, 21 and 25 x 10(3) were common to tachyzoites of all strains, however, there were significant quantitative and qualitative differences in the antigen profiles, indicating a moderate degree of strain specific polymorphism in tachyzoite antigens. We found no specific association between antigenic variation and strain virulence. Comparison of tachyzoite and bradyzoite antigens from homologous strains (RRA, DEG, ME49) confirmed the existence of stage specific antigens and demonstrated a conserved antigen profile among bradyzoites. PMID- 11027787 TI - The effects of subcurative doses of chloroquine on Plasmodium vinckei petteri gametocytes and on their infectivity to mosquitoes. AB - The effects of subcurative doses of chloroquine on rodent and human Plasmodium transmission to the mosquito have been studied by several authors who showed a short-term (12 h) enhancement of gametocyte infectivity by the drug, restricted to chloroquine-resistant strains, and a long term (4-6 days) enhancement of gametocytogenesis of chloroquine-sensitive strains of Plasmodium chabaudi. We investigated both short- and long-term effects of chloroquine on Plasmodium vinckei petteri, a chloroquine-sensitive rodent Plasmodium strain. Chloroquine treatment reduced the index of gametocytogenesis to 73% (5 mg/kg) and 55% (2.5 mg/kg) of controls, on day 6 post-infection (p.i.). The reduction was statistically significant with 5 mg/kg chloroquine. However, the reduction of gametocyte numbers did not affect the transmission capabilities of the strain. Our experiments showed that doses of 1 mg/kg chloroquine had no effect on the oocyst counts, 12 h post-administration to mice. A statistically non-significant 61% reduction of oocyst numbers was observed in mosquitoes fed on mice treated with 5 mg/kg chloroquine. The effect of 5 mg/kg chloroquine administration on the infectivity of gametocytes to mosquitoes fed 1 h post-treatment was also investigated. An overall 41% reduction of oocyst numbers was observed. This immediate effect was statistically significant in 73% of the mice. These results are consistent with the hypothesis that the short-term enhancing effect of chloroquine on transmission is restricted to the drug-resistant strains of Plasmodium. PMID- 11027788 TI - Molluscicidal activity of 2-hydroxy-3-alkyl-1,4-naphthoquinones and derivatives. AB - In the search for new molluscicidal agents we tested the activity of lapachol and other 2-hydroxy-3-alkylnaphthoquinones possessing nitrogenated alkyl chains, against the snail Biomphalaria glabrata. Lapachol, isolapachol and nor-lapachol showed strong molluscicidal activity against the adult snail (LD(90)<10 ppm) and significant toxicity against snail egg masses (LD(90)<0.2 ppm). As lapachol is easily extracted, and the derivatives can be synthesised without any difficulty, large-scale synthesis and field tests can be conducted, with a view to large scale molluscicidal programs. PMID- 11027789 TI - The microneme protein MIC4, or an MIC4-like protein, is expressed within the macrogamete and associated with oocyst wall formation in Toxoplasma gondii. AB - The expression and localisation of MIC4, or an immuno-cross reacting MIC4-like protein, was examined in the enteric forms of Toxoplasma gondii using immunocytochemistry. In addition to being located within the micronemes of the merozoites, MIC4 or the MIC4-like protein was present within the macrogamete and was associated with the developing oocyst wall. The macrogamete is characterised by two types of structurally distinct wall forming bodies (WFB1 and 2). However, by immuno-electron microscopy, it was possible to identify two populations of dense granules (WFB1) which appear to form sequentially during macrogamete development. The first granules to form (WFB1a) stained positively with anti-MIC4 and were followed by MIC4 negative granules (WFB1b). During oocyst wall formation, the WFB1a and b sequentially released their contents onto the surface with WFB1a material forming an anti-MIC4 positive outer veil, while the WFB1b forms the electron dense outer layer of the oocyst wall. The inner layer was formed by WFB2. Thus, for the first time, it was possible to identify two populations of dense granules (WFB1a and b) involved in the formation of different parts of the oocyst wall. It was not possible to analyse the contents of macrogametes by western blot to unequivocally identify the antigen recognised by the polyclonal antisera as MIC4. PMID- 11027790 TI - Introduction PMID- 11027791 TI - P300 as a clinical assay: rationale, evaluation, and findings. AB - Use of the P300 event-related brain potential (ERP) as a clinical assay is reviewed and assessed by comparing its distribution qualities with normative biomedical testing data from published studies. The coefficient of variation statistic was calculated for P300 data and a variety of clinical testing data. P300 amplitude and latency variability was found to be highly comparable and sometimes superior to routinely employed biomedical assays. These results are discussed in terms of how to control inter-group ERP variability and the application of normative P300 data in clinical settings. PMID- 11027792 TI - The objective assessment of amnesia in dissociative identity disorder using event related potentials. AB - Assessment of amnesia in Dissociative Identity Disorder (DID) typically relies on self-report, the veracity of which cannot often be independently verified. Memory in DID was therefore assessed using an objective method that involved event related potentials (ERPs) as well as indirect behavioral measures of memory, and that provided statistically supported assessments for each participant. Four participants who met DSM-IV criteria for DID participated in an ERP memory assessment task, in which words learned by one identity (identity A) were then presented to a second identity (identity B). All four participants - tested as identity B - produced ERP and behavioral evidence consistent with recognition of the material learned by identity A. While it would be premature to generalize all cases of DID, the results suggest that there may be reasons to question the veracity of reports by individuals who meet diagnostic criteria for DID on the basis of a structured clinical interview. PMID- 11027793 TI - P300 in family studies of schizophrenia: review and critique. AB - The published studies using P300 event-related potential (ERP) as a vulnerability marker in schizophrenia have been summarized and reviewed. Several factors across sibling, offspring, and familial studies appear to contribute to the variegated findings with respect to the utility of P300 as a vulnerability marker for schizophrenia. These include how subject samples are defined in terms of their genetic loading for schizophrenia, sample size, ERP methodology, and the relationship of P300 measures to neuropsychological test outcomes. It was concluded that the P300 holds promise as a vulnerability marker for schizophrenia, if used in conjunction with other neuroimaging and neuropsychological testing methods. PMID- 11027795 TI - Left temporal deficit of P300 in patients with schizophrenia: effects of task. AB - P300 is often, but not always, observed to be more reduced over left than right temporal lobes in patients with schizophrenia. The possibility that task differences contribute to the inconsistency in the literature was explored in this study. ERPs were collected from 17 right-handed men with schizophrenia (DSM IIIR) and 11 right-handed healthy male community controls, performing three auditory oddball tasks - respond to a target tone by: (1) counting; (2) pressing a response button with the right index finger; or (3) pressing a response button with the left index finger. Although patients with schizophrenia had smaller and later P300 amplitudes than controls, they did not have smaller P300s over the left temporal scalp (T3) than over the right (T4). P300 recorded over the left (C3) and right (C4) motor cortices indicated sensitivity to responding hand, with greater negativity being associated with contralateral button pressing. Failure to find P300 asymmetry is not related to the presence or absence of a button pressing task, or the hand used for button pressing. Rather, P300 asymmetry may be related to structural neuroanatomical asymmetries. PMID- 11027794 TI - The application of cognitive event-related brain potentials (ERPs) in language impaired individuals: review and case studies. AB - There is a substantial body of basic research that has utilized ERPs to investigate the neurological basis of cognition. This research has, in turn, led to the development of practical applications of cognitive ERPs in patient populations. In particular, recent work has focused on the development of ERP based assessment measures for the neuropsychological assessment of dyslexia and language impairments secondary to stroke. This review describes the innovative assessment methods program (IAMP), an initiative to utilize ERPs for a neuropsychological assessment of patients who cannot be evaluated by traditional methods. The success of this program has demonstrated that ERPs can be used to reliably evaluate an individual's reading and speech comprehension abilities, independent of behavioral and speech production impediments. In contrast to traditional neuropsychological assessment, these ERP methods can discern the cognitive strategies used by an individual to perform a task. PMID- 11027796 TI - Identifying a multivariate endophenotype for substance use disorders using psychophysiological measures. AB - This investigation examined how reduced amplitude of the P300 event-related potential (elicited from a visual oddball task) can be used together with an electrodermal response modulation measure (indexing the ability to inhibit responsivity to a temporally predictable aversive stimulus) to identify adolescents at especially high risk to develop substance dependence. One hundred and twenty-nine 17-year-old boys were divided into groups characterized as low risk (high amplitude P300 and good electrodermal modulation), high risk (reduced amplitude P300 and poor modulation), or intermediate risk (a high or good score on one measure and a low or poor score on the other). P300 amplitude and electrodermal modulation were uncorrelated. High-risk boys had 4-6 times more alcohol dependence than intermediate or low-risk boys and 2-3 times more nicotine dependence. Performance on an antisaccade eye-tracking task in which participants directed their gaze in a direction opposite to target movement was related to electrodermal modulation but not P300 amplitude. The results from all three psychophysiological measures together suggest that the neural circuits affecting P300 amplitude and electrodermal response modulation are different and that poor electrodermal response modulation may reflect an inhibitory control deficit mediated by the frontal lobes. PMID- 11027797 TI - Neuroelectric assessment of HIV: EEG, ERP, and viral load. AB - The effects of the human immunodeficiency virus (HIV) infection on the central nervous system function were studied with electroencephalographic (EEG) and auditory event-related brain potentials (EPRs) in patients infected with HIV and unaffected young adult control subjects (n=10/group). All subjects were assessed once every 15 min for four trial blocks at the same time of day to assess EEG/ERP changes with time on task-induced fatigue. Spectral analysis was applied to the pre- and post-stimulus EEG segments. ERP values were evaluated with respect to group differences for component amplitude and latency measures. Spectral analysis demonstrated that HIV patients evinced greater pre-stimulus delta power over frontal areas compared to control subjects, and less post-stimulus spectral power for the delta, theta, and alpha bands over the central/parietal areas. P300 amplitude was smaller, and latency was marginally longer for the HIV patients compared to control subjects. P300 latency correlated positively with increases in the patient HIV viral load. Time-on-task generally did not affect EEG or ERP measures for either group other than contributing to an overall decrease in neuroelectric responsivity. Group spectral power effects were consistent with differences in arousal/fatigue level. P300 group differences were consistent with declines in cognitive capability, and P300 latency increased with increased viral load. HIV infection negatively affected central nervous system function as measured by EEG and cognitive ERPs in a manner that suggests decreased arousal and increased fatigue in HIV patients. PMID- 11027798 TI - Introducing GAVI and the Global Fund for Children's Vaccines. PMID- 11027799 TI - Inactivated hepatitis A vaccine booster given >/=24 months after the primary dose. AB - We investigated what happens with the immune response when people come back for their booster dose of inactivated hepatitis A vaccine later than the recommended time of 6-12 months after the primary dose. We recruited a group of 124 travellers who received either the primary doses of Havrix 720 (two doses) or of Havrix 1440 (one dose) >/=24 months before study entry. They received a booster dose of Havrix 1440 and blood was drawn 1 month later. As a control group, we recruited a group of 125 travellers who followed a recommended schedule with a primary dose at month 0 and a booster dose at months 6-12. For both study groups, the GMTs increased dramatically and similarly upon the booster immunisation. Although significantly more late travellers (32%) had lost detectable antibodies than controls (11%) before administration of the booster dose, all these subjects showed an anamnestic response to the booster dose. Delaying the booster dose up to 66 months after primary vaccination did not seem to influence the immunogenicity of the booster dose. However, the recommended 6-12-month interval remains if detectable antibody titers are to be warranted constantly. PMID- 11027801 TI - Vaccines and variant CJD. PMID- 11027800 TI - Immunogenicity and reactogenicity of the Biken acellular pertussis vaccine in young adults. AB - To assess the reactogenicity and immunogenicity of the Biken acellular pertussis vaccine (Pa) following administration of a single vaccine dose to young adults with or without a history of prior pertussis immunization, 104 healthy, male and female adults without primary pertussis immunization were enrolled in Mainz (former West Germany; "not previously pertussis vaccinated", N-PPV-group); in parallel, 103 adults with a history of having received >/=four doses of a combined diphtheria-, tetanus-toxoid, whole-cell pertussis vaccine (DTwP) were enrolled in Magdeburg (former East Germany; "previously pertussis-vaccinated", PPV-group). Large areas of redness (>20 mm) were seen in 2.9%/1.0% of subjects in the N-PPV/PPV group, and a large swelling was seen in 6.8%/1.0%, respectively. As compared to baseline concentrations, antibody titers to PT and FHA were 77 (PT)/64-(FHA) (N-PPV group) and 94-(PT)/126-(FHA) fold (PPV group) higher. The study vaccine was safe and induced infrequent and mostly mild, local and general symptoms that all resolved spontaneously; it was highly immunogenic in adults, whether or not they had been previously vaccinated with DTwP. PMID- 11027802 TI - Pneumococcal surface adhesin A antibody concentration in serum and nasopharyngeal carriage of Streptococcus pneumoniae in young African infants. PMID- 11027803 TI - Oral, intrarectal and intranasal immunizations using CpG and non-CpG oligodeoxynucleotides as adjuvants. AB - We have previously demonstrated that synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs (CpG ODN) are potent adjuvants in mice when delivered by intramuscular, intranasal and subcutaneous routes. Herein, using tetanus toxoid (TT) as a model antigen in BALB/c mice, we compared the ability of CpG ODN to induce mucosal and systemic humoral immune responses when antigen was delivered by three different routes: intrarectal, intranasal and oral. Results showed differences in immune responses with the three routes and also revealed that non-CpG "control" ODN had adjuvant effects when used at mucosal sites. This was unexpected since non-CpG ODN do not have such immunostimulatory effects in vitro or after parenteral immunization. These findings were further investigated after oral delivery of a killed influenza vaccine on its own as well as combined with TT and hepatitis B surface antigen. Our findings demonstrate that with mucosal delivery, there is a Th2 immunostimulatory effect associated with the phosphorothioate ODN backbone, and that the presence of CpG motifs shifts this towards a Th1 response. PMID- 11027804 TI - Immunization with recombinant LD1 antigens protects against experimental leishmaniasis. AB - The genes, ORFF and BT1 (previously ORFG), are part of the multigenic LD1 locus on chromosome 35 which is frequently amplified in Leishmania. BT1 encodes a biopterin transporter, while the function of the ORFF gene product is unknown, but it is localized to the nucleus. We show here that immunization of mice with recombinant ORFF and BT1 proteins, individually, or in combination, conferred partial protection against challenge with Leishmania donovani. Protection correlated with the production of antigen-specific antibodies and in vitro splenocyte proliferation. Thus, these antigens can be potential vaccine candidates against visceral leishmaniasis. PMID- 11027806 TI - Immunogenicity and reactogenicity of the combined hepatitis A and B vaccine in young adults. AB - The combination of hepatitis A virus (HAV) and hepatitis B virus (HBV) vaccinations can offer convenience, increased compliance and cost saving. We have studied the immunogenicity, reactogenicity and safety of combined hepatitis A and B vaccination in young adults (16-35 years old). Eighty healthy young adults were divided into two random groups. One group received the combined hepatitis A and B vaccine (HAB) in one arm while the other group was administered concomitant hepatitis A and B vaccines (HAV + HBV) in the right and left arms, respectively. The immunogenicity, reactogenicity and safety were assessed after each dose in both the groups. In local symptoms, the percentage of the combined HAB group was lower than the HAV + HBV group, and the general symptoms were noted in approximately 30% of each group without any significant difference. No serious adverse effects were noted. All the subjects were seropositive for antibody to hepatitis A virus (anti-HAV) after one dose of vaccine, and remained seropositive after three doses in both groups. The seropositive rate for antibody to hepatitis B surface antigen (anti-HBs) was significantly higher (84%) in the combined HAB group than the concomitant HAV + HBV group (62%), (p<0.05) after dose two, and all the subjects were seropositive (100%) after the third dose. The GMTs of anti HAV and anti-HBs were not significantly different between groups 1 and 2 (p>0.1) except in month 6 when the GMT of anti-HBs was higher in HAB group (p=0.0039). The combined HAB vaccine was found to be safe, well tolerated and had less local symptoms in young adults. The immunogenicity and reactogenicity were similar to the concomitant HAV + HBV vaccines. PMID- 11027805 TI - Minimum estimated incidence in Japan of anaphylaxis to live virus vaccines including gelatin. AB - We have previously found that most occurrences of anaphylaxis to live virus vaccines are caused by gelatin present in the vaccines as a stabilizer. After we published the evidence for the role of gelatin in anaphylaxis, vaccine manufacturers in Japan began to eliminate gelatin from live virus vaccines. In the present study, we tried to estimate its incidence before the gelatin elimination was started. Physicians and vaccine manufacturers submitted serum samples from children with anaphylaxis to measles, mumps, rubella or varicella vaccine to National Institute of Infectious Diseases (NIID) for 3 years from April 1994 to March 1997. Specific IgE to gelatin was assayed at NIID or two manufacturers by the CAP and ELISA methods. There were 44 children with life threatening severe anaphylaxis (airway obstruction or anaphylactic shock) during the 3-year period, 41 of whom had anti-gelatin IgE. There were 64 children with mild anaphylaxis (without airway obstruction); 62 had anti-gelatin IgE. There were 100 children with only systemic cutaneous signs; 81 had anti-gelatin IgE. The estimates for the incidence of the severe anaphylaxis in 1994-1996 are: 6.84, 7.31, 4. 36, and 10.3 cases per million doses of gelatin-containing measles, rubella, mumps, and varicella vaccines, respectively. PMID- 11027807 TI - Absence of chicken myelin basic protein residues in commercial formulations of MMR vaccine. AB - Several preparations of MMR vaccines and their progenitor monovalent vaccine bulks produced by two different manufacturers were examined serologically for the presence of chicken myelin basic protein (MBP) residues. The products were challenged against several commercial preparations of anti-hMBP antisera that reacted positively with the control MBP preparations of human and chicken origins. There was no evidence of the presence of MBP components in MMR vaccines or their progenitor vaccine bulks as shown by the reactivity profiles of the antibody preparations against control and test antigens. PMID- 11027808 TI - Chimeric classical swine fever viruses containing envelope protein E(RNS) or E2 of bovine viral diarrhoea virus protect pigs against challenge with CSFV and induce a distinguishable antibody response. AB - Three chimeric classical swine fever virus (CSFV)/bovine viral diarrhoea virus (BVDV) full-length DNA copies were constructed, based on the infectious DNA copy of the CSFV vaccine strain C. The antigenic region of E2 and/or the complete E(RNS) gene were replaced by the analogous sequence of BVDV II strain 5250. Viable chimeric virus Flc11, in which E(RNS) was replaced, was directly recovered from supernatant of SK6.T7 cells transfected with full-length DNA. Viable chimeric virus Flc9, in which E2 was replaced, resulted in recovery of virus only when SK6.T7 transfected cells were maintained for several passages. However, no virus could be recovered after replacement of both E(RNS) and E2, even after 10 cell passages. Both Flc9 and Flc11 grow in swine kidney cells (SK6), stably maintain their heterologous BVDV sequences and, as assessed by monoclonal antibody typing and radio-immunoprecipitation assays, express their heterologous proteins. Flc9 showed a slower growth rate on SK6 cells than Flc11 and wild-type Flc2 virus. Replacement of E(RNS) or E2 of C-strain-based chimeric viruses did not alter cell tropism compared to wild-type C-strain virus for SK6 and FBE cells. Both Flc9 and Flc11 induced E2 or E(RNS) antibodies, which could be discriminated from those induced after wild-type virus infection, even after repeated vaccination. Furthermore, pigs were completely protected against a lethal CSFV challenge. These results indicate the feasibility of introduction of marker antigens in a live-attenuated marker C-strain vaccine for CSFV. PMID- 11027809 TI - Salmonella typhimurium as a basis for a live oral Echinococcus granulosus vaccine. AB - A live attenuated Salmonella typhimurium vaccine candidate, LVR01, was constructed by introducing a null deletion into the aroC gene of the parental canine S. typhimurium isolate, P228067. LVR01 was used to orally deliver to the canine immune system a fatty acid binding protein (FABP) from Echinococcus granulosus (EgDf1), as a fusion protein with fragment C (TetC) of tetanus toxin. Immunization studies demonstrated that live LVR01 is well tolerated by orally vaccinated dogs. There was no detectable shedding of the vaccine strain in the faeces 2 days after immunization. Humoral antibody responses were observed against Salmonella, TetC and EgDf1. Cellular responses were consistently detected against Salmonella and TetC. A cellular response against EgDf1 was also seen in a proportion of the LVR01 vaccinated dogs. We propose S. typhimurium LVR01 as a carrier for recombinant antigens and a vector for the construction of multivalent oral vaccines for dogs. PMID- 11027810 TI - Hepatitis A and B in children and adolescents--what can we learn from Puglia (Italy) and Catalonia (Spain)? AB - Viral hepatitis remains a major contributor to the global disease burden. Mass immunisation strategies against hepatitis B have been adopted by more than 90 developing and industrialised countries. Countries with low hepatitis A endemicity are experiencing cyclical outbreaks and an epidemiological shift, with larger numbers of individuals at risk of infection at an older age, resulting in increased morbidity. The high cost of outbreaks in these countries has made immunisation strategies cost-effective. The development of a vaccine against hepatitis A and a combined vaccine against hepatitis A and hepatitis B offers potentially exciting opportunities for a preventative approach in areas of both low and high endemicity. Existing mass immunisation programmes against hepatitis B will facilitate the adoption of joint strategies illustrated by the examples of Puglia (Italy) and Catalonia (Spain). PMID- 11027811 TI - An experimental infection with classical swine fever in E2 sub-unit marker vaccine vaccinated and in non-vaccinated pigs. AB - The clinical and virological protection induced by an E2 sub-unit marker-vaccine against Classical Swine Fever (CSF) was examined during an experimental infection in vaccinated and non-vaccinated pigs. Forty-five pigs were equally distributed over three adjacent pens of an isolation unit, there was only indirect (airborne) contact between pigs in the different pens. In pen 3 all pigs were vaccinated twice with 4 weeks interval. Pigs in pens 1 and 2 were not vaccinated. Two weeks after booster vaccination, one randomly selected pig in the middle pen was experimentally inoculated with CSF virus. After the initial virus spread in the infected pen, all pigs in the non-vaccinated adjacent pen were infected. In the vaccinated pen, seven out of 14 pigs became infected during the experiment. Survival analysis showed that virus transmission by direct and indirect contact was significantly (p<0.001) delayed in vaccinated pigs as compared to non vaccinated pigs. In the non-vaccinated pens over 40% of the pigs died and typical clinical signs were noticed. In the vaccinated pen no mortality and no clinical symptoms were observed. Although double vaccination with an E2 sub-unit marker vaccine was able to prevent the clinical course of the disease it was unable to prevent infection through indirect contact. This finding combined with the slow serological response after vaccination will complicate the possible use of the vaccine in emergency vaccination programmes. PMID- 11027812 TI - Safety and immunogenicity of NYVAC-JEV and ALVAC-JEV attenuated recombinant Japanese encephalitis virus--poxvirus vaccines in vaccinia-nonimmune and vaccinia immune humans. AB - A controlled, randomized, double-blind clinical trial evaluated whether two attenuated recombinant poxviruses with identical Japanese encephalitis virus (JEV) gene insertions, NYVAC-JEV and ALVAC-JEV, were safe and immunogenic in volunteers. Groups of 10 volunteers distinguished by vaccinia immune status received two doses of each vaccine. The vaccines appeared to be equally safe and well tolerated in volunteers, but more reactogenic than licensed formalin inactivated JE and placebo vaccines given as controls. NYVAC-JEV and ALVAC-JEV vaccine recipients had frequent occurrence of local warmth, erythema, tenderness, and/or arm pain after vaccination. There was no apparent effect of vaccinia immune status on frequency or magnitude of local and systemic reactions. NYVAC JEV elicited antibody responses to JEV antigens in recipients but ALVAC-JEV vaccine poorly induced antibody responses. However, NYVAC-JEV vaccine induced neutralizing antibody responses only in vaccinia-nonimmune recipients while vaccinia-immune volunteers failed to develop protective antibodies (5/5 vs. 0/5 seroconversion, p<0.01). These data suggest that preexisting immunity to poxvirus vector may suppress antibody responses to recombinant gene products. PMID- 11027813 TI - Induction of mucosal and systemic antibody specific for SeMF3 of Streptococcus equi by intranasal vaccination using a sucrose acetate isobutyrate based delivery system. AB - Streptococcus equi causes equine strangles, a highly contagious disease of the upper respiratory tract. The antiphagocytic surface protein SeM is strongly immunogenic and evokes mucosal and systemic antibodies during convalescence. The present study investigated the potential of sucrose acetate isobutyrate (SAIB); a high viscosity excipient that provides controlled release of biologically active substances, to enhance antibody responses following intranasal immunization of horses with a 108 a.a. peptide of SeM (SeMF3). SeMF3-SAIB was administered intranasally to each of the 11 adult horses on days 0 and 28. A second group of seven horses was vaccinated with SeMF3 alone. SAIB enhanced the mucosal and systemic immunogenicity of SeMF3, whereas SeMF3 by itself stimulated only a shortlived mucosal IgA and no systemic response. Moreover, nasal mucosal responses of horses immunized with SeMF3-SAIB were qualitatively and quantitatively similar to those observed in convalescent horses and involved similar linear epitopes of SeM. Epitope analysis also suggested that the nasal response was different from that observed in serum. A booster response was obtained after the second vaccination. These results suggest that SAIB has potential as a vehicle for intranasal immunization of horses with antigenic peptides. PMID- 11027814 TI - Protection against hydatid disease induced with the EG95 vaccine is associated with conformational epitopes. AB - This paper describes attempts to map the location of host-protective epitopes of a recombinant vaccine antigen by assessing the ability of truncated regions of the antigen to elicit protective immune responses in sheep. Sheep were immunised with three truncated regions (EG95-1, EG95-2 and EG95-3) of the hydatid vaccine antigen, EG95. These regions overlapped each other and corresponded to amino acids 1-70 (EG95-1), 51-106 (EG95-2) and 89-153 (EG95-3) of the full length recombinant protein. Each region elicited antibody which reacted with the parent antigen, although these reactivities were a small proportion of the level of reactivity generated by immunisation with the full length antigen. Antisera raised against each of the truncated proteins reacted with the native parasite antigen. In vaccination and parasite challenge trials in sheep, none of the truncated regions elicited significant protection against challenge infection or antibody which was lethal to the parasite in vitro. Antibodies from sheep immunised with the combination of all three overlapping truncations elicited a comparatively low but significant level of lysis of the parasite in vitro. These antigens did not inhibit anti-EG95 antibody reactivity with EG95 nor did they inhibit in vitro oncosphere killing induced by anti-EG95 antibodies. These results indicate that the major part of the immune response induced by EG95 vaccination is directed against conformational epitopes and that the host protective epitope(s) is/are conformational. PMID- 11027815 TI - Plasmodium falciparum AMA-1 erythrocyte binding peptides implicate AMA-1 as erythrocyte binding protein. AB - The role of AMA-1 during merozoite invasion has not yet been determined. However, reported experimental evidence suggests that this protein can be used, in particular as erythrocyte-binding protein, since, Fab fragments against this protein are able to block merozoite invasion. Using a previously described methodology, eight peptides with high binding activity to human erythrocyte, scattered along the different domains and having around 130 nM affinity constants, were identified in the Plasmodium falciparum AMA-1 protein. Their binding activity was sialic acid independent. Some of these peptides showed homology with the erythrocyte binding domains of one of the apical organelle protein family, MAEBL, identified in rodent malarial parasites. One of these peptides shares amino acid sequence with a previously reported B-cell epitope which induces antibodies to block parasite growth. The critical residues were identified for erythrocyte binding conserved peptides 4313 (DAEVAGTQYRLPSGKCPVFG), 4321 (VVDNWEKVCPRKNLQNAKFG), 4325 (MIKSAFLPTGAFKADRYKSH) and 4337 (WGEEKRASHTTPVLMEKPYY). All conserved peptides were able to block merozoite invasion of new RBC and development, suggesting that these peptides are involved in P. falciparum invasion. PMID- 11027816 TI - Introduction of the haemagglutinin transmembrane region in the influenza virus matrix protein facilitates its incorporation into ISCOM and activation of specific CD8(+) cytotoxic T lymphocytes. AB - The gene encoding the influenza virus A matrix (MA) protein was cloned into the bacterial expression vector pMalC with and without the sequence encoding the transmembrane region of the haemagglutinin (HA). With the resulting recombinant proteins, immune stimulating complexes (ISCOM) were prepared. The MA protein with the hydrophobic anchor region (rMAHA) associated more efficiently with ISCOM than the unmodified MA protein (rMA). A B-lymphoblastoid cell line (B-LCL) was lysed by an autologous CD8(+) cytotoxic T lymphocyte (CTL) clone specific for the MA protein after incubation with rMAHA-ISCOM but not after incubation with rMA, rMAHA, rMA-ISCOM or empty ISCOM. The B-LCL was also lysed by the CTL clone after incubation with empty ISCOM mixed with the respective MA proteins. Incubation of ISCOM with the rMAHA protein proved to be the most efficient in this respect. Addition of the proteasome inhibitors lactacystin or clasto-lactacystin beta lactone to the B-LCL incubated with rMAHA-ISCOM or the MA proteins mixed with empty ISCOM dramatically decreased the lysis by the CD8(+) CTL clone. These results indicate that the addition of a hydrophobic anchor to hydrophilic proteins in combination with ISCOM facilitates their entry in the MHC class I processing and presentation pathway. This may be an attractive approach for the development of subunit vaccines aiming at the induction of CTL-mediated immunity. PMID- 11027817 TI - Modelling antibody response to measles vaccine and subsequent waning of immunity in a low exposure population. AB - We analysed measles antibody data from 1141 vaccinated Canadian children aged 4 16 years to investigate factors associated with seropositivity and magnitude of prevailing antibody titres. We used a mixed Bernoulli/log-gamma model which takes account of the possibility of primary vaccine failures, of censored observations due to the lower detection limit of antibody assays and of a potentially skewed distribution of the logarithm of titres. While controlling for all significant variables, we observed that measles antibody titres decline at a mean rate of 5.6% per annum (approximate 95% CI; 3.3-7.7) equivalent to a half life of 12 years. Further, we noted a detectable linear relationship with respect to age at immunisation such that the mean increase in measles antibody titre of an infant who responds to vaccination gained by delaying vaccination from 12 to 18 months of age is 11.1% (approximate 95% CI; 4.4-18.3). PMID- 11027818 TI - Effect of immunological adjuvant combinations on the antibody and T-cell response to vaccination with MUC1-KLH and GD3-KLH conjugates. AB - A year ago we described a comparison of 19 immunological adjuvants for their ability to augment antibody and T-cell responses against vaccines containing two cancer antigens, GD3 ganglioside and MUC1 peptide, covalently attached to keyhole limpet hemocyanin (KLH). As in our previous experience, the saponin fraction QS 21 was the most potent single adjuvant but several other adjuvants also had potent adjuvant activity. Induction of an immune response against cancer antigens is generally difficult because these antigens are autoantigens. To get maximal benefit from the adjuvant component of cancer vaccines we have now tested whether combinations of the optimal adjuvants induced an improved immune response compared to QS-21 alone. Since over the intervening year a new semi-synthetic saponin adjuvant (GPI-0100) containing the dodecylamide derivative of hydrolyzed naturally-occurring saponins had become available, this was tested as well. Twelve different adjuvant combinations and GPI-0100 were compared for their ability to augment (1) antibody responses against GD3 and MUC1 and (2) T-cell responses against GD3, MUC1 and KLH. GPI-0100 and five adjuvant combinations were superior to QS-21 alone for induction of IgM and IgG antibodies against MUC1 and/or GD3: QS-21 plus bacterial nucleotide CpG, QS-21 plus monophosphoryl lipid A (MPL), QS-21 plus non-ionic block copolymer CRL-1005, QS-21 plus Titermax and Titermax plus CpG. Antibody responses were documented both by ELISA against purified antigens and by FACS for cell surface reactivity. There was no evidence for T-cell immunity against GD3 or MUC1. The antibody responses against GD3 and MUC1 were, however, strongly correlated with IFN-gamma release and DTH against KLH. These results demonstrate that combinations of immunological adjuvants are able to augment antibody and T-cell responses to these conjugates beyond that attainable with QS-21 alone, and again confirm the absolute necessity of potent adjuvants or adjuvant combinations for optimal immunogenicity with conjugate vaccines. PMID- 11027819 TI - An immunoglobulin G based chimeric protein induced foot-and-mouth disease specific immune response in swine. AB - Epitopes containing the residues 141aa-160aa and 200aa-213aa from foot-and-mouth disease (FMD) serotype O1K HK type FMDV VP1 were joined to a swine immunoglobulin G single heavy chain constant region (scIgG), creating a novel chimeric protein, named F1-scIgG. In this study, inoculation with F1-scIgG induced both FMD virus neutralizing antibody response and T cell response in swine. Antisera from these F1-scIgG-inoculated swine protected suckling mice against 1000 lethal dose 50 (1000LD(50)) FMD challenge. F1-scIgG-inoculated swine were also fully protected against 50LD(50) FMD virus challenge. The present study demonstrates the clear potential for viral epitopes linked with self-Ig in novel FMD vaccine design. PMID- 11027820 TI - Salivary antibody response to vaccination with meningococcal A/C polysaccharide vaccine in previously vaccinated and unvaccinated Gambian children. AB - Development of salivary antibodies at the age of 4 or 5 years to group A and C meningococcal polysaccharides (MenA/C PS) was studied among Gambian children, who had received MenA/C conjugate or PS vaccine in infancy. There was also a control group of 64 age matched children. IgG, IgA, and secretory Ig concentrations were measured by enzyme immuno assay. MenA/C PS vaccine induced antibodies both in previously vaccinated and unvaccinated children. The previous vaccination had not induced long lasting IgA-mediated memory. IgA antibodies were secretory, and most of IgG was serum derived. The IgG salivary response seen was similar to the serum response. PMID- 11027821 TI - Accessory cell defect in unresponsiveness of neonates and aged to polysaccharide vaccines. AB - T independent antigens elicit antibody responses in the absence of carrier specific T helper cells but require signals from accessory cells (macrophages and dendritic cells) or specific cytokines. They are further subdivided into TI-1 and TI-2 categories based on the ability of TI-1 but not TI-2 antigens to elicit immune responses from neonates. Most bacterial polysaccharides including the pneumococcal polysaccharide vaccines belong to the TI-2 class. It is hypothesized that defects in accessory cell function play a critical role in the failure of neonates to respond to such TI-2 antigens. Immune responses to these TI-2 stimuli are also reduced in the aged, also due to a quantitative deficiency in accessory cells. Agents that can stimulate accessory cell function may provide an alternative strategy to improve the immunogenicity of the polysaccharide vaccines in the neonates and the aged. PMID- 11027822 TI - A single dose sub-unit vaccine protects against pneumonic plague. AB - In this study, the protection afforded against aerosolised Yersinia pestis by injection of a single dose of an alhydrogel-adsorbed sub-unit vaccine has been compared with that given by an existing killed whole cell vaccine licensed for human use. The sub-unit vaccine, prepared by admixing F1 antigen derived from a Y. pestis cell culture supernatant with recombinant V antigen derived from an E. coli cell lysate, fully protected an outbred strain of mouse against exposure to 10(6) CFU of virulent plague organisms (10(4) mouse lethal doses, MLD). In contrast, the whole cell vaccine provided only 16% protection against the same level of challenge. Furthermore, sub-unit vaccinees were able to clear the bacteria from their lungs post-challenge whereas bacteria were cultured from the lungs of a surviving KWC vaccinee post-challenge. In killed whole cell vaccinees, physiologically significant levels of IgG to F1 only were detectable and the levels of F1-specific IgG in serum and in broncho-alveolar washings were significantly lower (p<0.05) compared with sub-unit vaccinees. In sub-unit vaccinees, an IgG titre to the F1 and V antigens was detected in serum where it was significantly higher (p<0.05) compared with broncho-alveolar washings suggesting that, at the time of challenge, protection is attributable mainly to the combined circulating IgG titre to the F1 and V sub-units. The enhanced protective efficacy of this sub-unit vaccine administered as a single dose compared with an existing vaccine has been demonstrated in an outbred animal model of pneumonic plague. PMID- 11027823 TI - Induction of lymphocyte recruitment in the absence of a detectable immune response. AB - Lymphocyte recruitment from blood into the lymph node is thought to be initiated by the presence of antigen. In this study, we have used lymphatic cannulation in sheep to demonstrate that the adjuvant ISCOMATRIX can induce dramatic lymph node activation in the absence of antigen. Consistent patterns of node shutdown (decreased output) and cell recruitment (increased output) with minimal blast cell responses were observed indicating that an antigen-specific immune response is not required. Production of IL-6, IL-8 and IFN-gamma, and the transient presence of red blood cells and neutrophils in the efferent lymph were associated with changes in efferent lymph cell trafficking. These early events may facilitate the screening of low frequency antigen-specific cells for binding to antigen and the subsequent amplification of the immune response. PMID- 11027824 TI - Oral immunization with size-purified microsphere beads as a vehicle selectively induces systemic tolerance and sensitization. AB - Oral administration of antigens has long been recognized as a method to prevent or delay the onset of diseases associated with untoward immune responses to self and non-self antigens. Although oral administration of antigens offers a convenient way to induce systemic tolerance, its therapeutic potential has been seriously limited by the fact that it requires repeated feeding of a large amount of antigens and that it may deteriorate ongoing autoimmune diseases when autoantigens are employed. We have previously shown that orally administered poly D,L-lactic acid (PDLLA) microspheres containing an antigen were selectively distributed to Peyer's patches (PP) and systemic lymphoid tissues according to their diameter and then released the antigen over a long period of time. We now report that a single dose of intragastric immunization with a PDLLA microsphere 7 10 micrometer in diameter and containing 2 mg of OVA was as effective as 100 mg of water soluble OVA to suppress OVA-specific IgG and DTH response. This was associated with a large increase of Interferon-gamma production by PPT cells stimulated with an antigen and a small increase in secretory IgA specific to OVA. In contrast, administration of an antigen encapsulated in microspheres 3-4 microm in diameter led to an enhanced OVA-specific IgG response and no significant increase in OVA-specific secretory IgA. Thus, by utilizing microspheres of an appropriate diameter as a vaccination vehicle, we were able to selectively induce both systemic tolerance and sensitization by oral ingestion of single low dose of an antigen. PMID- 11027825 TI - Characterization of mucoadhesive microspheres for the induction of mucosal and systemic immune responses. AB - In the present study, mucoadhesive polymer-dispersed microspheres (MS) were examined as a potential mucosal vaccine carrier. A major focus of the study was aimed at directly assessing the influence of antigen release and persistence in the mouse small intestine for the induction of mucosal and systemic immune responses. BALB/c mice were immunized with various forms of MS containing chicken egg ovalbumin (OVA) by administration into the duodenum. No detectable anti-OVA immune responses were observed following the administration of OVA alone or that of MS without mucoadhesive polymer (MS-0). MS-10 containing 10% mucoadhesive polymer rapidly released OVA and hardly induced anti-OVA antibody responses in either serum or fecal extracts. In contrast, MS-8 and MS-6 (with 8 and 6% mucoadhesive polymer) showed controlled release of OVA, which elicited strong OVA specific IgG and IgA responses in serum and fecal extracts, respectively. Additionally, the strongest immune responses were induced in mice immunized with MS-8, which had both the optimal release-profile of OVA and the longest persistence in the small intestine. These findings indicate that antigen movement in the small intestine is an important factor and that appropriate microsphere forms with mucoadhesive polymers might be useful candidates as mucosal vaccine carriers. PMID- 11027826 TI - Stabilization of vaccines: to be or not to be. PMID- 11027827 TI - Competition among streptococcus pneumoniae for intranasal colonization in amouse model PMID- 11027828 TI - The suppressors of cytokine signaling (SOCS) proteins: important feedback inhibitors of cytokine action. AB - While positive effectors of cytokine signaling pathways are relatively well defined, negative regulation can be just as important but is poorly understood. The recently discovered suppressor of cytokine signaling (SOCS) family of proteins has been implicated in the negative regulation of several cytokine pathways, particularly the receptor-associated tyrosine kinase/signal transducer and activator of transcription (AK/STAT) pathways of transcriptional activation. Biochemical studies revealed that inhibition can occur via a variety of mechanisms. SOCS proteins bind to tyrosine-phosphorylated residues of target proteins via their SH2 domains, then inhibit JAK activity through their N terminal domains, and are thought to induce degredation of bound molecules through a conserved SOCS-box motif that interacts with the proteasome. SOCS protein expression is induced by a wide variety of cytokines with each member displaying varying kinetics of induction. Gene modification studies in mice have demonstrated that SOCS-1 has a clear role in the negative regulation of interferon-gamma signaling, while other SOCS family members have also been shown to be involved in the regulation of T cell, growth hormone, and erythropoietin signaling systems. PMID- 11027829 TI - Functional characterization of the two alternative promoters of human p45 NF-E2 gene. AB - OBJECTIVE: The transcription factor NF-E2, a heterodimeric protein complex composed of p45 and small Maf family proteins, is considered crucial for the proper differentiation of erythrocytes and megakaryocytes in vivo. We report the results of studies aimed at understanding the regulatory mechanisms controlling p45 gene expression in erythroid cells. MATERIALS AND METHODS: Human p45 mRNAs have two alternative isoforms, aNF-E2 and fNF-E2, and these isoforms are transcribed from the alternative promoters. We investigated lineage-specific expression of both isomers in human erythroid and megakaryocytic cells by reverse transcriptase polymerase chain reaction or Northern blot analysis. For functional characterization of both promoters, plasmids in which reporter genes were placed under the control of a series of truncated or mutated promoter fragments were transfected to human hematopoietic cell lines. RESULTS: When CD34(+) cells isolated from human cord blood were induced to unilineage erythroid or megakaryocytic differentiation in liquid suspension culture, both transcripts, although barely detected at day 0, were induced in both erythroid and megakaryocytic cultures. fNF-E2 mRNA was found to be more abundant in erythroid cells than megakaryocytic cells at day 7 of culture. Although both isomers were expressed in human erythroid-megakaryocytic cell lines, megakaryocytic maturation with loss of erythroid phenotype induced by phorbol 12-myristate 13-acetate (PMA) resulted in exclusive downregulation of fNF-E2, suggesting that fNF-E2 promoter is more erythroid specific. Functional analysis of fNF-E2 promoter showed that the promoter is active only in erythroid-megakaryocytic cells and that the double GATA site in the proximal region is necessary for its efficient activity. CONCLUSION: These results suggest that GATA proteins, which govern the differentiation of erythroid lineage cells, are required for full promoter activity of the p45 gene. PMID- 11027830 TI - Interleukin-6 and other gp130-dependent cytokines selectively inhibit proliferation of macrophage-lineage hemopoietic progenitor cells. AB - OBJECTIVE: Hemopoiesis is regulated by cytokines with positive or negative effects on proliferation of lineage-committed or multipotent hemopoietic stem cells. We have investigated the roles of interleukin-6 and other gp130-dependent ligands on the proliferation of macrophage-lineage hemopoietic progenitor cells. METHODS: The responses of human and murine hemopoietic cells to combinations of cytokines involving interleukin-6 or related factors were assessed in short-term culture by clonogenic assay. RESULTS: Interleukin-6, leukemia inhibitory factor, and ciliary neurotrophic factor inhibited formation of colonies stimulated by macrophage colony-stimulating factor. These effects were dose dependent and selective for macrophage-lineage precursors. Progenitors from murine peripheral blood were inhibited by 37-93% in cultures containing interleukin-6 (11 experiments; median, 68%). Macrophage progenitors from murine bone marrow were also inhibited by interleukin-6 but were less sensitive (seven experiments; median, 48%). In cultures costimulated with leukemia inhibitory factor, peripheral blood and bone marrow progenitors were inhibited by 82% and 58%, respectively. Ciliary neurotrophic factor inhibited macrophage colonies by 66%. Multilineage bone marrow colony formation was not affected. In cultures of human bone marrow cells stimulated with macrophage colony stimulating factor and stem cell factor, interleukin-6 inhibited colony formation by 51-74%. Bone marrow colonies stimulated by granulocyte-macrophage colony stimulating factor were not inhibited by costimulation with interleukin-6. CONCLUSIONS: These results suggest a novel mechanism for the negative regulation of macrophage-lineage hemopoietic cells. They also demonstrate new properties of interleukin-6 and certain other gp 130-dependent ligands. PMID- 11027831 TI - Gelatin beads as platforms for targeting molecule and anti-Fas antibody: two major properties of cytotoxic T lymphocytes. AB - OBJECTIVE: Cytotoxic T lymphocytes (CTL) initially recognize target cells using the T-cell receptor (TCR), then strongly adhere to these cells by accessory molecules, and finally induce apoptosis by Fas ligand (FasL)/Fas or lyse by the granzyme/perforin system. We describe the development of gelatin beads carrying anti-tumor monoclonal antibody (mAb) and anti-Fas mAb mimicking the TCR and FasL, respectively. We hypothesized that these antibody-coated beads can be therapeutically utilized for the elimination of tumor cells. MATERIALS AND METHODS: We evaluated the cytotoxic activity of gelatin beads bearing CH11 (anti Fas mAb) after incubation with several human leukemia cell lines. Cytotoxic activities were measured using colorimetric DNA fragmentation assay and lactate dehydrogenase (LDH) release assay. RESULTS: We demonstrated that the cytotoxic effects of anti-Fas mAb were markedly enhanced by fixation on gelatin beads. Microscopic examination showed that the beads attached to the target cells and induced their apoptosis. These effects were enhanced further by adding tumor specific mAb. These in vitro properties of the beads were well reconstituted in the peritoneal cavity of mice. CONCLUSION: Although antibody-coated gelatin beads lack several important properties of natural CTL, such as differentiation, proliferation, and the functions of adhesion molecules, they mimic well the targeting and cytotoxic functions of natural CTL. Our findings suggest that antibody-carrying gelatin beads may be the first step toward the development of artificial CTL and can be applied, for example, to artificial dendritic and stroma cells for the development of novel biotherapeutic approaches. PMID- 11027832 TI - Ex vivo T lymphocyte expansion for retroviral transduction: influence of serum free media on variations in cell expansion rates and lymphocyte subset distribution. AB - OBJECTIVE: In the setting of allogeneic stem cell transplantation, suicide gene manipulated donor T cells that can be selectively inactivated in vivo would potentially allow optimal control of the GVL (graft-vs-leukemia)/GVHD (graft-vs host disease) balance. Retroviral T-cell transduction requires ex vivo cell expansion, which is often achieved by IL-2 and anti-CD3 stimulation. Traditionally, culture media for cell expansion are supplemented with fetal bovine serum (FBS) or human serum. While these sera promote cell growth and viability, they contain uncharacterized elements that may yield inconsistent results from batch to batch. Cell expansion in serum-free media would therefore be preferable. MATERIALS AND METHODS: We compared T-cell expansion rates in three commercially available serum-free culture media (X-VIVO 15, AIM-V, and Cellgro SCGM), with or without the addition of human serum (HS, 5%). We also aimed to evaluate how the in vitro expansion affected the composition of the various T cell subsets. Buffy-coats from four healthy donors were expanded for 21 days. The media were compared to standard RPMI 1640 medium, supplemented with HS (5%) or FBS (10%). For retroviral transductions, the LN vector carrying the neomycin- resistance gene was used in four additional donors. RESULTS: In our hands, X-VIVO 15 gave the highest rate of serum-free expansion (a median of 79-fold expansion, range 20-117). For serum-free expansion, activation with OKT3 for 21 days gave slightly higher expansion rates than a 5-day course (however, without statistical significance). When serum was added, this discrepancy was not seen. Cytokine analysis (IFN-gamma, IL-10, and IL-4) showed a distinct type1 cytokine pattern with elevated IFN-gamma levels during the whole period of culture. Flow cytometric analyses showed substantial inter-media, but also some inter-donor, variability in T-cell subset compositions. Transduction of cells with the LN vector and G418 selection resulted in a 14-fold increase (range 3-18) for serum free X-VIVO 15 based cultures. Cell phenotypes remained unchanged by the transduction procedure as compared to nontransduced cells. CONCLUSION: Among the tested serum-free media, X-VIVO 15 has shown to best support the in vitro expansion of T cells, resulting in equal percentages of CD4(+) and CD8(+) cells. These cells can easily be transduced and selected. There seem to be no significant benefits, regarding absolute cell numbers or T-cell subset compositions, with OKT3-stimulation for more than five days. The addition of low levels of HS increases the consistencies in the cell expansion rates for all media. PMID- 11027833 TI - Activation of mitogen-activated protein kinase through alpha5/beta1 integrin is required for cell cycle progression of B progenitor cell line, Reh, on human marrow stromal cells. AB - OBJECTIVE: Attachment to bone marrow (BM) stromal cells is crucial for the normal growth and development of B-cell progenitors (pro-B). However, the molecular mechanisms by which contact facilitates the proliferation of pro-B cells are not completely understood. This study was performed to investigate this interaction. MATERIALS AND METHODS: A model pro-B cell line (Reh) and a human BM stromal cell line (KM102) were used. Flow cytomery was used for cell cycle analysis. Western Blotting and immunoprecipitation were utilized to examine the levels of cyclin dependent kinase (cdk) and p27(Kip1). RESULTS: Attachment to both KM102 and normal BM stromal cells significantly promoted the growth of Reh cells. Pretreatment of Reh cells with anti-integrin beta1 or alpha5 monoclonal antibody (mAb), but not alpha4 or ICAM-1 mAb, abrogated this enhancement of proliferation. Furthermore, stroma attachment resulted in shortening of the G(1) phase of cell cycle, significant increases cdk2 activity, degradation of cdk inhibitor p27-GST protein, and decrease in levels of p27(Kip1) protein. In addition, solid-phase cross-linking of alpha5 via immobilized antibody also resulted in extracellular signal-regulated (ERK)-2 kinase phosphorylation, increase in cdk2 activity, decrease in levels of p27(Kip1) protein, and enhanced proliferation that was inhibited by treatment with PD98059, a specific ERK inhibitor. CONCLUSION: Integrin alpha5beta1-mediated stroma contact promotes the proliferation of B-cell progenitors through the activation of ERK-2, which in turn modulates cell cycle regulation machinery including induction of cdk2 activity and degradation of p27(Kip1) and contributing to acceleration of the G(1) phase of cell cycle progression. PMID- 11027834 TI - Alteration of c-mpl-mediated signal transduction in CD34(+) cells from patients with myelodysplastic syndromes. AB - OBJECTIVE: Megakaryocytic differentiation is frequently defective in patients with myelodysplastic syndromes (MDS). As underlying mechanisms, deregulated thrombopoietin receptor (c-mpl)-mediated signaling pathways have been suggested. This study therefore examined whether the impaired signaling in MDS and AML cells includes alterations of c-mpl itself or postsignaling events. METHODS: Bone marrow-derived CD34(+) cells from healthy donors, patients with MDS (RA, RAEB-T), and patients with AML after MDS were isolated by MACS. Expression of c-mpl cDNA was studied by RT-PCR. Thrombopoietin dependent activation of STAT proteins and MAP Kinase p42(erk-2)/44(erk-1) was analyzed by Western blot. RESULTS: Both splicing isoforms of c-mpl (c-mpl-p and c-mpl-k) were expressed in all of the CD34(+) cells examined. Analysis of the c-mpl cDNA revealed no sequence abnormality. We show c-mpl dependent activation of the transcription factors STAT3 and STAT5 as well as MAP Kinase p42(erk-2)/44(erk-1) in CD34(+) cells from healthy individuals. Cells derived from RA patients revealed low basal levels of phosphorylated STAT3 and STAT5 molecules. This phosphorylation was enhanced by stimulation with recombinant thrombopoietin (PEG-rHuMGDF). STAT1 failed to be activated by PEG-rHuMGDF in CD34(+) cells from healthy donors as well as from patients with MDS. In RAEB-T and AML M7 the constitutive expression levels of STAT1, 3, 5, and MAPK were markedly upregulated, resulting in a strong activation of STAT3 and 5 by PEG-rHuMGDF. Despite its high expression, the level of MAPK phosphorylation was not increased in RA or RAEB-T compared to the normal control, and was completely undetectable in AML M7. CONCLUSION: These results suggest that the defective megakaryopoiesis in MDS is not caused by a lack of c-mpl and that STAT3 and STAT5 may contribute to the malignant phenotype of the leukemic cells. PMID- 11027835 TI - Receptor for macrophage colony-stimulating factor transduces a signal decreasing erythroid potential in the multipotent hematopoietic EML cell line. AB - OBJECTIVE: To test the hypothesis that hematopoietic growth factors may influence lineage choice in pluripotent progenitor cells, we investigated the effects of macrophage colony-stimulating factor (M-CSF) on erythroid and myeloid potentials of multipotent EML cells ectopically expressing M-CSF receptor (M-CSFR). METHODS: EML cells are stem cell factor (SCF)-dependent murine cells that give rise spontaneously to pre-B cells, burst-forming unit erythroid (BFU-E), and colony forming unit granulocyte macrophage (CFU-GM). We determined BFU-E and CFU-GM frequencies among EML cells transduced with murine M-CSFR, human M-CSFR, or chimeric receptors, and cultivated in the presence of SCF, M-CSF, or both growth factors. Effects of specific inhibitors of signaling molecules were investigated. RESULTS: EML cells transduced with murine M-CSFR proliferated in response to M CSF but also exhibited a sharp and rapid decrease in BFU-E frequency associated with an increase in CFU-GM frequency. In contrast, EML cells expressing human M CSFR proliferated in response to M-CSF without any changes in erythroid or myeloid potential. Using chimeric receptors between human and murine M-CSFR, we showed that the effects of M-CSF on EML cell differentiation potential are mediated by a large region in the intracellular domain of murine M-CSFR. Furthermore, phospholipase C (PLC) inhibitor U73122 interfered with the negative effects of ligand-activated murine M-CSFR on EML cell erythroid potential. CONCLUSION: We propose that signaling pathways activated by tyrosine kinase receptors may regulate erythroid potential and commitment decisions in multipotent progenitor cells and that PLC may play a key role in this process. PMID- 11027836 TI - Immunophenotypic and functional characterization of CD33(+)CD34(+) cells in human cord blood of preterm neonates. AB - OBJECTIVE: To characterize CD33(+)CD34(+) cells, a major population in human cord blood (CB) CD34(+) cells of preterm neonates. MATERIALS: The proportion of CD33(+) cells was analyzed on CB CD34(+) cells from preterm and full-term neonates. CD33(+)CD34(+) cells were purified by cell sorting and analyzed on their clonogenic activity, proliferative activity in short-time liquid suspension culture, and GATA-2 mRNA expression by RT-PCR and Southern blot. RESULTS: The absolute numbers and proportion of CD34(+) cells in mononuclear cells inversely correlated with gestational age. CD33 was expressed on a majority of CB CD34(+) cells of preterm neonates but on only a minor population of them in full-term neonates. In addition, CD33 was dominantly expressed on CD38(-)CD34(+) cells or CD117(low)CD34(+) cells in CB of preterm neonates. CD33(+)CD34(+) cells of preterm cord blood had high proliferative and reproducible potentials compared with CD33(-)CD34(+) cells. CD33(+)CD34(+) cells as well as CD33(-)CD34(+) cells from preterm CB highly expressed GATA-2, in contrast to those from BM. CONCLUSIONS: These results suggest that CD33(+)CD34(+) cells, which are a major population in CB CD34(+) cells of preterm neonates, do not simply represent relatively mature myeloid lineage hematopoietic progenitor cells as those in adult BM CD34(+) cells, and may contain hematopoietic stem cells or primitive progenitor cells as in fetal liver. PMID- 11027837 TI - Increased expansion and differentiation of cord blood products using a two-step expansion culture. AB - OBJECTIVE: [corrected] The use of allogeneic cord blood (CB) products as a source of cellular support for patients receiving high-dose chemotherapy has been limited primarily to smaller children due to the low numbers of cells in a CB unit. Ex vivo expansion of CB cells has been proposed as a method to increase the number of cells available for transplantation. Following high-dose chemotherapy administration, we transplanted adult patients with CB expanded in static culture for 10 days, in DM containing stem cell factor (SCF), granulocyte colony stimulating factor (G-CSF), and megakaryocyte growth and development factor (MGDF). Patients achieved neutrophil engraftment in a median of 26 days (range 15 to 45). In an attempt to hasten the time to neutrophil engraftment, we developed a two-step culture system that results in increased expansion of total nucleated cells and further maturation of neutrophil precursors. MATERIALS AND METHODS: CD34(+) cells isolated from CB products were cultured for 7 days at 37 degrees C in 100-mL Teflon culture bags containing 50 mL of DM containing SCF, G-CSF, and MGDF (100 ng/mL). The cells were harvested from these bags after 7 days of incubation at 37 degrees C and transferred to 1-L Teflon bags containing 1 L of DM plus SCF, G-CSF, and MGDF. After a second culture period of 7 days, the cells were harvested, washed, and assayed for mature (granulocyte-macrophage colony forming cells [GM-CFC]) and primitive progenitor cells (high proliferative potential colony-forming cells [HPP-CFC]). RESULTS: The two-step cultures resulted in a median total nucleated cell expansion of 438-fold (range 286 to 952, N = 11); the original one-step cultures resulted in a median expansion of 98 fold (range 59 to 350, N = 5). Equivalent expansion of committed progenitor cells (GM-CFC) and primitive progenitor cells (HPP-CFC) was obtained. CD34(+) cells were expanded a median of 29-fold in the two-step cultures (N = 11). The two-step culture contained more mature neutrophil cells, by morphologic examination, than the one-step cultures, similar to ex vivo expanded peripheral blood progenitor cells (PBPC). CONCLUSION: The two-step ex vivo expansion conditions described for CB resulted in increased numbers of total nucleated cells, GM-CFC, HPP-CFC, and CD34(+) cells and morphologically resembled ex vivo expanded PBPC, which have been shown to provide more rapid neutrophil engraftment than unexpanded PBPC. We propose that the availability of increased numbers of expanded CB cells may result in more rapid engraftment of neutrophils following infusion to transplant recipients. PMID- 11027839 TI - The influence of diazepam on the electroencephalogram-evoked potential interrelation in rats. AB - Though being a sedative, diazepam increases beta-activity in the electroencephalogram (EEG). Diazepam also affects auditory evoked potentials (AEPs). We investigated if the effect of diazepam on AEPs could be ascribed to its beta-increasing effect. Eight rats received vehicle and diazepam counterbalanced. AEPs were ranked in ten categories according to the percentages of beta-activity in the pre-stimulus electroencephalogram (EEG). With vehicle, the P(29), N(51) and P(67) AEP components increased and cross-correlation coefficients (CCCs) declined with increasing beta-activity. With diazepam AEP components and CCCs remained unchanged. All AEPs with diazepam closely resembled the AEP of the lowest beta-category obtained with vehicle. We conclude that the effect of diazepam on AEPs can not be ascribed to its beta-enhancing effect. Diazepam disrupts the normal AEP-EEG relation such that diazepam-effects on AEPs seem to reflect the sedative effects of diazepam and not its beta-increasing effects. PMID- 11027838 TI - A prospective study of positive/negative ex vivo B-cell depletion in patients with chronic lymphocytic leukemia. AB - OBJECTIVE: Autologous peripheral blood stem cell (PBSC) transplantation is increasingly being used in patients with chronic lymphocytic leukemia (CLL). As the autografts are frequently contaminated with large numbers of tumor cells, we have prospectively investigated the feasibility and efficacy of ex vivo double purging of PBSC grafts in an open, nonrandomized, single-center phase I/II clinical study. MATERIALS AND METHODS: Twenty consecutive patients with poor-risk CLL underwent uniform stem cell mobilization with chemotherapy and granulocyte colony-stimulating factor (G-CSF). Double B-cell depletion of the harvested PBSC products was performed using immunomagnetic CD34(+) cell selection (Isolex300i Nexell, Irvine, CA) followed by a negative step with anti-CD19/20/23/37-labeled immunomagnetic beads. The purified PBSC were reinfused after myeloablative treatment with TBI/CY. RESULTS: A total of 25 separation runs was accomplished using collection products containing 3.4% (1.1-8.1) CD34(+) cells and 1.2% (0.1 42) CD19(+)CD5(+) CLL cells. After double selection, 33% (15-67) CD34(+) cells were recovered with a purity of 98.8% (89.1-99. 8). CLL cells were undetectable by high-resolution flow cytometry in 15 of 25 final products; median purging efficacy was 5 (4.1-6) log. The CD34(+) content of the 20 final grafts was 4.6 (2.2-6.5) x 10(6)/kg. Rapid and durable engraftment developed in all cases. With a median follow-up of 20 (6-29) months, 17 patients live in complete clinical remission, two have recurrent disease, and one patient died due to pulmonary embolism five months after transplant. Persistence of the leukemic clone on the molecular level was demonstrated by dot blotting with clone-specific CDR3 probes in an additional five patients. Serious or unexpected infectious complications did not occur. CONCLUSIONS: Positive/negative purging with the Isolex system allows preparation of highly purified CD34(+) fractions and up to six log of tumor cell depletion in patients with B-CLL and can be safety reinfused after myeloablative therapy without affecting hematopoietic engraftment. PMID- 11027840 TI - Plasma levels of 24S-hydroxycholesterol in patients with neurological diseases. AB - The brain is the exclusive or almost exclusive site of formation of 24S hydroxycholesterol and we have shown that the circulating level of 24S hydroxycholesterol is dependent upon the relation between cerebral production and hepatic clearance. In the present work we determined plasma levels of 24S hydroxycholesterol in patients with various neurological diseases. Eleven subjects with brain death occurring 6-10 h before collection of the plasma samples had markedly reduced circulating levels of 24S-hydroxycholesterol (-43%, P<0.001). Patients with advanced Alzheimer's disease and cerebral inflammatory diseases had slightly lower levels of 24S-hydroxycholesterol in plasma when compared to matched controls. Patients with acute ischemic stroke, multiple sclerosis and primary brain tumors had levels not significantly different from those of controls. The conditions leading to reduced plasma levels of 24S hydroxycholesterol had no significant effect on plasma levels of another side chain oxidized oxysterol, 27-hydroxycholesterol. Except for conditions characterized by very marked destruction of the central nervous system, different severe neurological diseases seem to have relatively small effects on the flux of 24S-hydroxycholesterol from the brain. PMID- 11027841 TI - The effect of streptozotocin-induced diabetes on cell proliferation in the rat dentate gyrus. AB - Recent evidence has shown an association between diabetes mellitus and deficits in learning and memory. However, the mechanism by which cognitive abilities are impaired in diabetes has not been identified. The dentate gyrus of the hippocampus plays a significant role in spatial learning and memory. Studies in rodents show that learning tasks enhance neurogenesis in the dentate gyrus of the adult hippocampus. To investigate whether cognitive deficits in diabetes may be related to alterations in hippocampal neurogenesis, we measured the number of 5 bromo-2'-deoxyuridine (BrdU)-positive cells, an indicator of cell proliferation, in the dentate gyrus in an animal model of diabetes. Streptozotocin-induced diabetes produced a dramatic decrease in cell proliferation in the dentate gyrus as compared to controls. The results from this study suggest a potential role for alterations in neurogenesis in the cognitive decline observed in diabetes mellitus. PMID- 11027842 TI - Topography and topology of the teleost telencephalon: a paradox resolved. AB - Analysis of vasculature in the telencephalic pallium of a teleost allows the considerable depth of the sulcus externus, which lies at the lateral extent of the ependymal attachment, to be appreciated. The depth of this sulcus is compelling evidence for a simple eversion process (an outfolding of the pallial wall of each hemisphere) during telencephalic development in all ray-finned fishes that is not complicated in teleosts by secondary migration of pallial cell groups. A simple eversion process is known to occur in some ray-finned fishes with relatively simple telencephalic cytoarchitecture but has been disputed in teleosts based on the pattern of olfactory tract projections. A resolution to the conflicting hypotheses of pallial relationships across ray-finned fishes and in comparison with other craniate radiations is presented here, based on a re examination of hodological and histochemical data mandated by this sulcal anatomy. PMID- 11027843 TI - Effects of hypothyroidism on insulin-like growth factor-I expression during brain development in mice. AB - Hypothyroidism has devastating consequences on brain development. While the mechanisms that mediate these effects are not known, several lines of evidence suggest that a reduction in insulin-like growth factor-I (IGF-I) expression and/or action has a role. To assess whether reduced IGF-I expression and/or actions mediates the brain pathology of congenital hypothyroidism, we induced hypothyroidism by treating pregnant mice and lactating dams with 0. 1% propylthiouracil (PTU) in drinking water. Control and PTU-treated pups were sacrificed on postnatal day (P) 7, 10 and 14, and IGF-I mRNA expression was assessed in the cerebral cortex and cerebellum by ribonuclease protection assay. To control for mRNA loading, the signal of IGF-I protected bands was normalized to those for cyclophillin. IGF-I mRNA expression in hypothyroid animals was decreased significantly in cortex at P10 and P14 (42 and 60%, respectively). In the cerebellum, IGF-I mRNA expression was down-regulated at all ages studied, but the decrease was only statistically significant at P7 (31% decreased). We conclude that hypothyroidism alters IGF-I expression in the developing brain. Furthermore, we speculate that IGF-I plays a role in mediating some thyroid hormone actions during brain development. PMID- 11027844 TI - Angiotensinogen and angiotensin converting enzyme gene polymorphisms and the risk of bipolar affective disorder in humans. AB - A possible participation of the renin-angiotensin system (RAS) components with mood disturbances has been suggested in both animal and pharmacological models. In this cross-sectional study, we examined the association between functional polymorphisms in the angiotensin converting enzyme (ACE) and angiotensinogen (AGT) genes in 115 bipolar affective disorder (BPAD) patients and 323healthy control subjects. The ACE I/D variant did not show any difference in allelic frequencies and genotypic distribution between the groups. In contrast, when studying the AGT M235T polymorphism we found that the M allele was more frequently observed in BPAD patients than in controls (chi(2)=6.766, d.f.=1, P=0.009). Using multivariate logistic models the strongest odds ratio resulted from a dominant genetic model (OR=3.0; CI (95%) 1.7-5.3] Our data suggest an association between the AGT M235 genotype and increased susceptibility for BPAD in these Brazilian patients. These findings are consistent with the hypothesis that the RAS system plays a role in regulating the mood PMID- 11027845 TI - Leptin injection into white adipose tissue elevates renal sympathetic nerve activity dose-dependently through the afferent nerves pathway in rats. AB - Recent studies suggested that leptin in white adipose tissue (WAT) affected the sympathetic out flow to several tissues. We examined whether elevations of renal sympathetic nerve activity (RSNA) and blood pressure (BP) could be observed by leptin injection into WAT in rats. Injections of leptin (10 and 100 ng/ml per kg) into WAT evoked the activation of RSNA dose-dependently. Circulating sympathetic nerve activators, such as leptin, insulin, glucose and lactate, were unchanged by any doses of leptin. In addition, BP was not affected by leptin injections during a 90 min experimental period. These data suggested that leptin activated the afferent nerves through the sensors in WAT, resulting in elevation of RSNA. PMID- 11027846 TI - Involvement of Ca2+/calmodulin-dependent protein kinase II in neurite outgrowth induced by cAMP treatment and serum deprivation in a central nervous system cell line, CAD derived from rat brain. AB - A central nervous system (CNS) cell line, CAD, is known to differentiate in the absence of serum. This cell line was found to differentiate by the treatment of cAMP. Expression of Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) was induced to about 2-fold or more on day 1, and was continued at a high level for 5 days after the exposure to differentiating conditions. Neurite extension was stimulated from day 1 and continued for 5 days, suggesting that CaM kinase II activity is correlated with neurite outgrowth. Of the four distinct isoforms (alpha, beta, gamma, and delta) of the kinase, the delta isoform was the major isoform in CAD cells. The splicing pattern of this isoform in the differentiated cells differed from that in undifferentiated cells, suggesting that expression of CaM kinase II is regulated during neural differentiation. PMID- 11027847 TI - Stutter-free and stutter-filled speech signals and their role in stuttering amelioration for English speaking adults. AB - This study examined the power of an exogenously generated stuttered speech signal on stuttering frequency when compared to an exogenously generated normal speech signal. In addition, we examined the specific components of the second speech signal, which might be responsible for the inducement of fluency in people who stutter. Eight males and two females who stuttered participated in this study. Experiment I involved meaningful speech: normal continuous speech, normal interrupted speech, stuttered continuous speech, and stuttered interrupted speech, whereas Experiment II involved vowels and consonants: /a/, /a-i-u/, /s/, /s-sh-f/. The results indicated that stuttered and normal speech signals were equally effective in reducing stuttering frequency. Further, the vowels were more powerful than consonants in inducing fluency for people who stutter. It is suggested that acoustic manifestations of stuttering, rather than a problem, may be a natural compensatory mechanism to bypass or inhibit the 'involuntary block' at the neural level. PMID- 11027848 TI - Both corticotropin releasing factor and neuropeptide Y are involved in the effect of orexin (hypocretin) on the food intake in rats. AB - Orexin (hypocretin) is a peptide that has been found to stimulate food intake in rats. However, we have recently demonstrated that orexin stimulates the release of corticotropin releasing hormone (CRF) which has been known to decrease the food intake. Therefore, we examined the mechanism of effect of orexin on food intake. Although the other appetite stimulating peptides; neuropeptide Y (NPY), agouti-related peptide (AGRP) and one of the growth hormone releasing secretagogue (GHRP-6) stimulated dose-dependently the food intake during 2 h in the early light period, orexin did not increase significantly the food intake. No significant increase was also observed during 2 h in the early dark period. However, pretreatment with alpha-helical CRF, an antagonist of CRF, or anti-CRF antiserum resulted in significant increase of food intake by orexin. Orexin stimulated feeding under these conditions was blocked by NPY Y1 receptor antagonist (1229U91). In an 8 h-fasting rat, anti-orexin serum decreased slightly the food intake. These results suggest that effect of orexin on the food intake may be complex because of orexin-CRF and orexin-NPY linkage. PMID- 11027849 TI - Continuous real-time measurement of extracellular ascorbate release in the rat striatum in vivo during forebrain ischemia-reperfusion. AB - Apart from its physiological role as a major antioxidant, ascorbate is highly concentrated in neuropils and ascorbate-mediated protection from excitotoxins has been demonstrated in vitro. Therefore, extracellular release of ascorbate during the early stage of ischemia-reperfusion was measured using a microdialysis electrode technique. One or two probes of the microdialysis biosensor were inserted into the rat striatum. One probe (n=16) was perfused with phosphate buffered saline (PBS) for continuous oxidative signal recording. A second electropolymerised probe inserted into the other side of the striatum was perfused with PBS containing ascorbate oxidase in six rats. Forebrain ischemia reperfusion was performed for 10min, followed by reperfusion for 60min. Ascorbate increased transiently during ischemia, and markedly to a maximum of 247.5+/-55. 8 microM from the baseline of 68.5+/-25.3 microM after reperfusion. The marked increase of extracellular ascorbate may be a marker of the early stage of reperfusion. PMID- 11027850 TI - The ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism does not confer protection against idiopathic Parkinson's disease. AB - The ubiquitin carboxy-terminal hydrolase L1gene (UCH-L1) has been implicated in the aetiology of Parkinson's disease (PD). A rare Ile93Met mutation in UCH-L1 in a German PD sib-pair has been reported. Recently, a S18Y (C54A) polymorphism in exon 3 of UCH-L1 was found to be under-represented in PD patients compared to controls. To test the reproducibility of this negative association, we conducted an allele-association study of the S18Y polymorphism in an Australian case control sample consisting of 142 PD cases and 142 closely matched control subjects. Genotypes were determined using polymerase chain reaction and RsaI restriction enzyme assay. Analysis revealed no significant difference between PD patients and controls for genotype or allele frequencies of the S18Y polymorphism. The frequency of the S18Y allele in Australian subjects is similar to that reported elsewhere. This study suggests that the S18Y polymorphism in UCH L1 does not influence the risk for developing PD. PMID- 11027851 TI - GM-1 ganglioside treatment reduces motoneuron death after ventral root avulsion in adult rats. AB - After ventral root avulsion, a large percentage of the wounded spinal motoneurons die within 2 weeks. Neuronal death is preceded by a series of morphological and physiological changes probably as a result of both apoptotic and necrotic mechanisms. Associated with neuronal death, inflammation develops in the wounded area, resulting in additional neuronal loss as well as the degeneration of fibers from different tracts of the ventral and lateral funiculi. In the present study we tested the neuroprotective and local anti-inflammatory effects of monosialoganglioside (GM-1) after an initial 150 mg/kg body weight dose, followed by the daily administration of 100 mg/kg body weight for 2 weeks. The results showed a statistically significant enhancement of surviving motoneurons which showed good morphological preservation. Also, GM-1 treatment reduced by less then a half, the number of degenerating fibers into the ventral and lateral funiculi. Taken together, our results indicate that the administration of GM-1 in high doses during the critical period of motoneuron death after avulsion is neuroprotective, and diminishes local inflammation. PMID- 11027852 TI - Lipopolysaccharide increases endogenous morphine levels in rat brain. AB - The present study was designed to determine whether whole body injection of lipopolysaccharide increases endogenous rat brain morphine levels. High performance liquid chromatography coupled to electrochemical detection and gas chromatography mass spectrometry reveal that the resting brain morphine level is 7.0+/-3.2ng/g wet weight. In a time dependent manner, intraperitoneal injection of lipopolysaccharide (10 microg), leads to an increase of brain morphine level with a peak reaching after 36h post injection. After 72h following the lipopolysaccharide-injection, morphine levels increased to 66.0+/-5.4ng/g brain wet weight (P<0.001). Furthermore, 96h of fasting also increased the brain morphine level to 44.0+/-3. 6ng/g of brain wet weight. These results suggest that rat brain morphinergic processes respond to organismic challenges after a significant latent period. PMID- 11027853 TI - Tuberomammillary nucleus activation anticipates feeding under a restricted schedule in rats. AB - We used FOS-immunoreactivity to map changes in the neuronal activity of brain nuclei related to the state of arousal, in rats under a restricted feeding schedule. Our main finding was the outstanding activation of the tuberomammillary nucleus 24h after a meal, and its steep deactivation, which was independent of actually having the meal. The time course of FOS activation and deactivation indicated a burst of tuberomammilary nucleus activity in close temporal relation with the increased locomotor activity shown by rats in anticipation of the next meal. PMID- 11027854 TI - Changes of cortical excitability in patients with upper limb amputation. AB - In our study we wanted to assess motor excitability in patients with upper limb amputation by means of transcranial magnetic stimulation (TMS). In 12 patients, TMS was applied using a paired pulse paradigm in order to test cortico-cortical excitability. Additional parameters of motor excitability like motor threshold and cortical silent period were also measured. Recordings from the amputated side were compared to the contralateral side and to healthy controls. We found a significant reduction of intracortical inhibition in forearm amputees and an enhancement of intracortical facilitation in upper arm amputees on the affected side. We conclude that after upper limb amputation, changes in the activity of intracortical interneuronal circuits appear in the affected hemisphere. These changes may depend on the level of amputation, and be the base of cortical reorganization. PMID- 11027855 TI - The relationships of nicotinamide adenine dinucleotide phosphate-d to nitric oxide synthase, vasoactive intestinal polypeptide, galanin and pituitary adenylate activating polypeptide in pigeon gut neurons. AB - The distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-d neurons and their relationship with nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), pituitary adenylate activating polypeptide (PACAP) and galanin (Gal) were examined in the gastrointestinal (GI) tract of the pigeon Columbia livia. NADPH-d-histochemistry, indirect immunofluorescence and confocal analysis were applied to cryosections. Western blot analysis was also applied on pigeon gut. NADPH-d neurons were found throughout the pigeon GI tract and they were evident in the myenteric, circular muscle and submucous plexuses. Positive varicose nerve fibres were also distributed within the longitudinal muscle layers and in the lamina propria of the mucosa. The stomach was the segment richest in positivities. The copresence VIP/Gal/NOS as well as PACAP/VIP were revealed in some NADPH-d-neurons. We suppose that the nitrergic nerve population of the pigeon GI tract belong to the muscle motility regulation as an inhibitory descending nerve pathway. Moreover the presence of VIP, Gal and PACAP in some NADPH-d-containing neurons enhances the inhibitory actions of these neurotransmitters whereas PACAP and Gal role is actually unknown. PMID- 11027856 TI - Prevalence of Giardia sp. Cryptosporidium parvum and Cryptosporidium andersoni (syn. C. muris) [correction of Cryptosporidium parvum and Cryptosporidium muris (C. andersoni)] in 109 dairy herds in five counties of southeastern New York. AB - A cross-sectional study was undertaken to determine the prevalence of Giardia sp. (G. duodenalis group), Cryptosporidium parvum and Cryptosporidium andersoni (C. muris) [corrected] in dairy cattle in three different age groups, and to evaluate the association of age and season with prevalence. One hundred and nine dairy farms, from a total of 212 farms, in five counties of southeastern New York volunteered to participate. On these farms, 2943 fecal samples were collected from three defined age groups. The farms were randomly assigned for sampling within the four seasons of the year. Each farm was visited once during the study period from March 1993 to June 1994 to collect fecal samples. Demographic data on the study population was collected at the time of sampling by interviewing the farm owner or manager. At collection, fecal samples were scored as diarrheic or non-diarrheic, and each condition was later related to positive or negative infection with these parasites. Fecal samples were processed using a quantitative centrifugation concentration flotation technique and enumerated using bright field and phase contrast microscopy. In this study, the overall population prevalence for Giardia sp. was 8.9%; C. parvum, 0.9%; and C. muris, 1.1%. When considering animals most at the risk of infection (those younger than 6 months of age) Giardia sp. and C. parvum was found in 20.1 and 2.4% of the animals, respectively. Giardia sp. and C. muris were found in all age groups. There was no significant seasonal pattern of infection for any of these parasites. PMID- 11027857 TI - Fatal toxoplasmosis in brown hares (Lepus europaeus): possible reasons of their high susceptibility to the infection. AB - Brown hares (Lepus europaeus) trapped in the countryside and domestic rabbits were experimentally infected with Toxoplasma gondii (K7 strain) oocysts. Hares (n=12) were divided into groups of 4 and infected with 10, 10(3) and 10(5) oocysts. Rabbits (n=12) were infected in the same way. The experimentally infected animals were monitored for 33 days after infection (p.i.). Most of the infected hares demonstrated behavioural changes, and all of them died between 8 and 19 days p.i. Three of the rabbits demonstrated only clinical changes related to the concurrent pasteurellosis. The typical pathological finding in the hares were haemorrhagic enteritis, enlargement and hyperaemia of mesenteric lymph nodes, splenomegaly and multiple miliary necrotic lesions in the parenchyma of the liver and other organs. Pathological changes in the rabbits were less pronounced than in the hares. In rabbit brains, tissue cysts of the T. gondii were found. The incidence of T. gondii antibodies both in the hares and the rabbits was first ascertained on day 7 p.i. On day 12 p.i., antibodies were already found in all the animals infected. Antibody titres in indirect fluorescence antibody test (IFAT) using the anti-rabbit conjugate were markedly higher in rabbits than in hares. In all hares, T. gondii was isolated post mortem from the liver, brain, spleen, kidney, lung, heart and skeletal muscles. Although T. gondii was also isolated in all rabbits, it was not always isolated in all their organs. In all hares, parasitemia was demonstrated on days 7 and 12 p.i. The percentage of rabbits with detected parasitemia was lower. In hares, a decrease in the numbers of leukocytes during the infection was observed. No such decrease was observed in the rabbits. The lymphocyte activity after the stimulation with non-specific mitogens showed significant differences between the hares and the rabbits even before the infection. After the infection, the hares infected with 10(3) and 10(5) doses and in rabbits infected with a 10(5) dose showed a decrease of lymphocyte activity. Rabbits infected with a 10(3) dose showed an increase of the lymphocyte activity. While in hares toxoplasmosis was an acute and fatal disease, the infection in rabbits had subclinical manifestations only and easily passed to a latent stage. The different courses of toxoplasmosis in the hare and the rabbit may be due to the differences in the natural sensitivity of the two species to the T. gondii infection or a negative impact of stress to the immune status of hares. PMID- 11027858 TI - Risk of babesiosis and anaplasmosis in different ecological zones of Santa Cruz Department, Bolivia. AB - A cross-sectional study was done of seroprevalence of Babesia bigemina, B.bovis, and Anaplasma marginale in cattle from eastern Bolivia, to characterise the risk of tick-borne disease in three ecological zones. Nineteen farms were sampled in the subtropical humid zone, 13 in the dry subtropical zone and nine in the lower western valleys of the Andean massif. The enzyme-linked immunosorbent assay was used. All three pathogens were widespread. For B. bovis, seroprevalences were high (75-78%) in the two subtropical zones which thus had low risk of disease from this infection; but the western valleys were endemically unstable with higher risk. For B. bigemina, seroprevalences were lower (24-57%) in the two subtropical zones and thus these areas were endemically unstable for disease from this infection. However, the seroprevalence of B. bigemina in the western valleys was too low (13%) for risk of disease in susceptible cattle to be considered high. For A. marginale, the seroprevalences in the two subtropical zones were low (19-32%) and very low (6%) in the western valleys suggesting all these zones were endemically unstable for anaplasmosis. Data for individual farms were analysed for risk of both forms of babesiosis; this showed low risk of disease in the subtropical humid zone, higher risk in the dry subtropical zone and variable risk in the western valleys. PMID- 11027859 TI - Use of a sandwich-enzyme-linked immunosorbent assay (SEA) for the diagnosis of natural Fasciola hepatica infection in cattle from Galicia (NW Spain). AB - An indirect-enzyme immune-linked immunosorbent assay (IEA) was compared to a sandwich-enzyme-linked immunosorbent assay (SEA) to diagnose fasciolosis in naturally infected cattle. By means of a rabbit polyclonal IgG antibody to F. hepatica antigens, captured circulating antigens were detected by an SEA technique. A total of 85.1% of the samples examined was considered positive by using the IEA-test, whereas this percentage was of 37.3% by the SEA test. Using the two enzymatic probes, only the 38.3% resulted positive to both immunoassay probes. Our data indicate that circulating antigen detection assay in combination with results of antibody detection may offer a more reliable diagnostic technique than classic coprologic ones, with results very appropriate for epidemiologic studies carried out on a wide area. PMID- 11027860 TI - Localization and migration of benzimidazole resistant and susceptible adult Cooperia curticei in the small intestine of sheep after treatment with thiabendazole. AB - Four groups of three lambs per group were experimentally infected with Cooperia curticei susceptible (two groups) or resistant (two groups) to benzimidazoles, and distributions of adult worms in the small intestine were studied. For each Cooperia isolate, one group was treated with thiabendazole (TBZ) (5 or 50mg/kg bodyweight) 28 days after infection. In the two untreated groups, the population of C. curticei were present from the second to the tenth meter of intestine from the pylorus with a maximum in the sixth meter for both isolates. After treatment with TBZ, the size of the resistant worm population did not significantly decrease but a large number of worms were found towards the proximal sections of the intestine. In contrast, the susceptible population was reduced by about 40% but the surviving worms remained at this same site of predilection after treatment. Measurements of the concentration of TBZ and 5OH-thiabendazole (5OH TBZ) in the intestinal segments do not indicate a clear relationship between the localization of worms and TBZ or 5OH-TBZ concentrations at least 12h after the anthelmintic treatment. The hypothesis of an enhanced expression of the mechanisms of resistance in the first few meters of small intestine is suggested. PMID- 11027861 TI - Experimental quantification of the transmission of Sarcoptes scabiei var. suis among finishing pigs. AB - In this study, the rate of S. scabiei var. suis transmission among finishing pigs was quantified in a contact transmission experiment. Forty piglets originating from a mange free farrow-to-finish herd were randomly allocated to three groups and one S. scabiei var. suis infested finishing pig was subsequently added to each of these groups. After 35 days, the three seeder pigs were removed from the groups and the remaining 40 pigs were re-allocated to five pens. Ear scrapings, to be examined for mites, were collected from each pig on days 1, 14, 28, 42, 56, and 84 of the experiment. Blood samples, to be tested for antibodies against S. scabiei, were collected from each pig on days 0, 14, 28, 42, 56, 70, 84 and 112 after the introduction of the seeder pigs. From the results of the ear scrapings and the blood samples the number of susceptible (not infested) and infested pigs was derived at the time of each sample collection and the number of new infestations in the intervals between the sample collections. From these data the infestation rate parameter beta (average number of new infestations per infested pig per day) was estimated by use of a Generalised Linear Model (GLM) and accordingly, beta was estimated at 0.056 (95% CI: 0.037-0.085) infestations per infested pig per day.Next, by use of beta, the transmission of S. scabiei was simulated in a population of 100 finishing pigs for 100 days after the introduction of a single infested pig. For this purpose, 500 simulations were done. The 90% confidence interval of the number of infested pigs at day 100 ranged from 12 to 88 (median: 63). It was concluded that transmission of S. scabiei among finishing pigs is slow. Due to the presumed lower contact rate between sows as compared to finishing pigs, it is anticipated that transmission of S. scabiei among sows will even be slower than among finishers These findings are of particular interest for the development of surveillance programmes for S. scabiei free herds. PMID- 11027862 TI - Control of fleas on naturally infested dogs and cats and in private residences with topical spot applications of fipronil or imidacloprid. AB - Thirty-four flea-infested dogs and cats residing in 20 homes in Tampa, FL were randomly placed into 1 of 2 treatment groups during the summer of 1997. Pets were treated topically with either 10.0%w/v imidacloprid or 10%w/v fipronil spot-on on day 0, then once for every 28-30 days for 90 days. Flea populations were assessed in the environment using an intermittent-light trap, while pet flea burdens were assessed using visual area counts. A single application of imidacloprid was 95.3 and 97.4% effective in reducing flea populations on pets at 7 and 28 days, respectively. A single application of fipronil was 97.5 and 97.0% effective in reducing flea populations on pets at the same time points. Following 3 monthly applications of either imidacloprid or fipronil, flea burdens on pets were reduced by 99.5 and 96.5%, respectively. In addition, flea numbers in the in-home environment were reduced by 99. 0 and 98.6% in homes, where pets were treated with imidacloprid or fipronil, respectively. PMID- 11027863 TI - Chlorfenapyr ear tags to control Haematobia irritans (L.) (Diptera: Muscidae) on cattle. AB - The efficacy of ear tags containing 30% chlorfenapyr (total tag weight=13g) to control natural Haematobia irritans (L.) infestations was evaluated for Holstein heifers in Rafaela, province of Santa Fe, Argentina. A group of heifers (TG) was treated with two ear tags (one tag per ear). A control group (ACG) was maintained in a paddock adjacent to the TG paddock and, a distant control group (DCG) was maintained 700m apart from the other groups. From day 4 to day 98 after treatment, H. irritans infestations of ACG were significantly higher (P<0.05, test of Kruskal-Wallis) than the corresponding infestation of TG, but significantly lower than infestation in DCG, probably due to the proximity of TG. The chlorfenapyr control period, with an efficacy higher than 90% to reduce horn fly populations, lasted for 9 weeks when TG infestation was compared to fly numbers in ACG, but increased to 12 weeks in comparison to DCG. The results of this study show that ear tags impregnated with chlorfenapyr are a useful alternative to pyrethroids and organo-phosphate compounds for horn fly control. PMID- 11027864 TI - Dermatitis associated with "hypopodes" in a horse: the first case reported in Italy. AB - The authors report, for the first time in Italy, a case of dermatitis of "hypopodes" origin in a horse. The hypopodes are a particular nymphal stage of mites of the suborder Astigmata. The "hypopus" is non-feeding, lacks a mouth and has a ventral suctorial plate with suckers and conoids for attaching itself to insects as a mode of dispersal. Some of these larval stages can enter into the hair follicles and into the subcutaneous layers causing lesions similar to mange. There are few reports of dermatitis in horses specifically associated with the presence of hypopodes and these have been attributed to an allergic response to the body fluid of crushed hypopial stage. It may be that infestations of these nymphal stages are widespread, but the nature of the condition is not yet clear; therefore, they may be underdiagnosed. PMID- 11027865 TI - Spectroscopic study of 1,2-indandione. AB - This paper describes the spectroscopic study of the new fingerprint reagent, 1,2 indandione, using nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS). When 1,2-indandione is dissolved in methanol, NMR and MS show formation of a hemiketal similar to that proposed for DFO when it is dissolved in methanol. In contrast to DFO where reactivity with amino acids appears to be enhanced by hemiketal formation with alcohol solvents, the reactivity of 1,2 indandione towards amino acids is diminished by hemiketal formation. Alcohols should be avoided in 1,2-indandione formulations. PMID- 11027866 TI - A rapid method to detect dried saliva stains swabbed from human skin using fluorescence spectroscopy. AB - Saliva on skin is important in forensic trace evidence. If areas where saliva is present can be outlined, this may lead to DNA analysis and identification. This study describes a rapid and non-destructive method to detect dried saliva on the surface of the skin by fluorescence spectroscopy. Eighty-two volunteers deposited samples of their own saliva on the skin of their ventral forearm. A control sample of water was deposited at three different sites on the contralateral arm. Saliva and water control were then allowed to air-dry. Swab samples were taken from dried saliva and control sites and were dissolved in 0.1M KCl solution. Emission spectra were obtained from the solution and were characterized by a principal maximum at 345-355nm with excitation at 282nm. The fluorescence emission intensity was greater than background readings obtained from the control swab site in 80 of 82 volunteers (approximately 97.6%). The fluorescence profile of saliva samples were similar to those obtained from aqueous samples of pure amylase and tryptophan, an endogenous fluorophore in alpha-amylase. The presence of an emission peak at 345-355nm with excitation at 282nm could provide a strong presumptive indication of saliva deposition. PMID- 11027867 TI - Synthesis of 2,3- and 3,4-methylenedioxyphenylalkylamines and their regioisomeric differentiation by mass spectral analysis using GC-MS-MS. AB - 3,4-methylenedioxyamphetamine (MDA) derivatives are increasingly abused central nervous system stimulants with neurotoxic properties. In recent years a number of controlled substance analogs (designer drugs) with high structural variety reached the illegal market making their identification an arduous task. The underivatized compounds give very similar or even virtually identical electron impact mass spectra containing mainly intense C(n)H(2n+2)N(+) immonium ions. Using tandem mass spectrometry (MS-MS) the additional structural information contained in the collision induced dissoziation (CID) mass spectra of molecular ions using electron impact (EI) and especially chemical ionization (CI) allowed an unequivocal differentiation of 18 studied regioisomeric 1 (methylenedioxyphenyl)-2-propanamines and 1-(methylenedioxyphenyl)-2-butanamines. Further synthetic methods are presented for 1-(3,4-methylenedioxyphenyl)-N-propyl 2-butanamine, N-isopropyl-1-(3,4-methylenedioxyphenyl)-2-butanamine and four 1 (2, 3-methylenedioxyphenyl)-2-butanamines. N-alkylated 1-(3, 4 methylenedioxyphenyl)-2-butanamine compounds (e.g. MBDB) are also known to be abused psychoactive agents (entactogenes) without the sympatomimetic effects of the 3,4-methylenedioxyamphetamines. PMID- 11027868 TI - Histologic appearance of fractured thyroid cartilage and surrounding tissues. AB - Histological appearances of the fractured superior horns (SH) of the thyroid cartilage and their surrounding tissues were reviewed, with particular reference to signs of vital origin of the fracture. Twenty-nine autopsies with either histories or findings indicating asphyxial neck compression, with a total of 39 fractured SHs, and three autopsies with history of suicidal jump from height with a total of two SH fractures, were examined. Fifteen autopsies with finding of 19 artefactually fractured SHs while removing neck organs at autopsy were used as controls. In the cases of neck compression and victims of jump from height haemorrhages, retraction of fractured SH fragments with invagination or squeezing of the perichondrium, contraction bands, contraction band necrosis and 'opaque fibres', in the adjacent muscle fibres could be seen. Fibrin deposition and/or leucocytic reaction were noted in cases where circumstances of death indicated prolonged death struggle (as in incomplete hanging, resuscitation or homicidal neck compression). However, retraction of fractured fragments and invagination of perichondrium between artefactually fractured SH fragments were observed in both control cases and cases with antemortem trauma to the larynx. Many of these histological findings, including haemorrhages and fractures had not been evident at gross examination. We conclude that histological examination of SHs may not only uncover macroscopically overlooked injuries but also may facilitate the clarification of an injury's vital origin. PMID- 11027869 TI - Magnetic resonance imaging screening in women at genetic risk of breast cancer: imaging and analysis protocol for the UK multicentre study. UK MRI Breast Screening Study Advisory Group. AB - The imaging and analysis protocol of the UK multicentre study of magnetic resonance imaging (MRI) as a method of screening for breast cancer in women at genetic risk is described. The study will compare the sensitivity and specificity of contrast-enhanced MRI with two-view x-ray mammography. Approximately 500 women below the age of 50 at high genetic risk of breast cancer will be recruited per year for three years, with annual MRI and x-ray mammography continuing for up to 5 years. A symptomatic cohort will be measured in the first year to ensure consistent reporting between centres. The MRI examination comprises a high sensitivity three-dimensional contrast-enhanced assessment, followed by a high specificity contrast-enhanced study in equivocal cases. Multiparametric analysis will encompass morphological assessment, the kinetics of contrast agent uptake and determination of quantitative pharmacokinetic parameters. Retrospective analysis will identify the most specific indicators of malignancy. Sensitivity and specificity, together with diagnostic performance, diagnostic impact and therapeutic impact will be assessed with reference to pathology, follow-up and changes in diagnostic certainty and therapeutic decisions. Mammography, lesion localisation, pathology and cytology will be performed in accordance with the UK NHS Breast Screening Programme quality assurance standards. Similar standards of quality assurance will be applied for MR measurements and evaluation. PMID- 11027870 TI - High-speed spectroscopic imaging for cancellous bone marrow R(2)* mapping and lipid quantification. AB - In this work an interleaved multiple-gradient-echo chemical shift imaging (IMGE CSI) technique was designed, implemented and evaluated at 1.5 and 4T for high resolution lipid quantification and R(2)* measurement in-vivo. The method is analogous to echo planar CSI but utilizes conventional gradient echoes, exploiting the principle of spectroscopic bandwidth extension by interleaving temporally offset gradient-echo trains. It is shown that IMGE-CSI is able to measure true fat volume fraction in oil/water mixtures with high accuracy, not possible with Dixon-type methods which approximate the spectrum as consisting of only two spectral components. Correlation of the CSI- derived volume fractions with volumetry afforded r(2) > 0.99 with a slope of 0.98. The method is shown to be able to quantify regional variations in bone marrow composition in vivo with a spatial resolution of 2.5 x 2.5 x 5 mm(3.) R(2)* was obtained by multi-line spectral curve fitting. For the measurement of R(2)* in cancellous bone marrow the method is shown to agree well with time-domain fitting techniques but is superior in instances where the marrow has both hematopoietic and fatty constituents. Finally, excellent inter-scan reproducibility (1% coefficient of variation for global means and medians) was achieved, yielding r(2) = 0.98 of the test-retest correlation for three scans in four test subjects. In conclusion, IMGE-CSI is found to enable highly accurate lipid quantification and measurement of cancellous bone marrow R(2)* at spatial resolutions and scan times typical of standard clinical protocols. PMID- 11027871 TI - Subtle white matter volume differences in children treated for medulloblastoma with conventional or reduced dose craniospinal irradiation. AB - Medulloblastoma is the most common malignant brain tumor in children, and approximately seventy percent of average-risk patients will achieve long-term survival. Craniospinal irradiation (CSI), combined with chemotherapy and surgery, is currently the mainstay of treatment but places children who survive at risk for serious neurocognitive sequelae. These sequelae are intensified with a younger age at treatment, greater elapsed time following treatment, and an increased radiation dose. Many newer treatment approaches have attempted to address this problem by reducing the dose of the CSI component of radiation therapy while maintaining the current survival rates. This study evaluates longitudinal MR imaging during therapy to assess the impact of the two CSI doses (conventional [36 Gy] and reduced [23.4 Gy]) on normal appearing white matter volumes (NAWMV) evaluated in a single index slice. Twenty-six children and young adults at least three years of age enrolled on an institutional protocol for newly diagnosed, previously untreated primary medulloblastoma had at least four MR examinations over a minimum nine month period following CSI. These serial volumes were evaluated as a function of time since CSI in three analyses: 1) all subjects, 2) subjects stratified by age at CSI, and 3) subjects stratified by CSI dose. The first analysis demonstrated that medulloblastoma patients treated with CSI have a significant loss of NAWMV in contradistiction to normally expected maturation. Stratifying the patients by age at CSI found no significant differences in the rate of NAWMV loss. The final analysis stratified the patients by CSI dose and revealed that the rate of NAWMV loss was 23% slower in children receiving reduced-dose. Serial quantitative MR measures of NAWMV may provide a neuroanatomical substrate for assessing functional impact of CSI on normal brain function following treatment for medulloblastoma. PMID- 11027873 TI - Diagnostic utility of tumor vascularity on magnetic resonance imaging of the breast. AB - The objective of this study was to examine the relation of tumor vascularity on magnetic resonance imaging (MRI) with differential diagnosis malignant from benign lesions and tumor invasiveness in breast carcinoma. Forty-nine patients with breast cancer or benign lesion (median 49 yrs) were examined with dynamic MRI. Scanning of the entire breast was performed at 1.5 T with a three dimensional fast spin echo sequence, using an original polarity altered spectral and spatial selective acquisition (PASTA) technique for fat suppression. Subsequently 0.1 mmol/Kg Gd-DTPA was administered and 3-6 images were obtained. The presence or absence of intratumoral, marginal and peritumoral vascularity on MRI was recorded. The excised specimen was histopathologically examined for the size of lesion, the presence and extent of local invasion. Tumor size on MRI correlated closely with the size at morphologic examination (r = 0. 96). Intratumoral (p = 0.04), marginal (p = 0.05) and peritumoral vascularity (p = 0.01) were less common in benign than in malignant lesions. Among the latter, intratumoral (p = 0.01) and marginal (p = 0.03) vascularity were more common in invasive carcinomas than in DCIS. In the subset of invasive carcinomas (n = 31); however, the tumors exhibiting intratumoral vascularity were markedly larger (p = 0.03). The presence of intratumoral and marginal vascularity on MRI can help predict both the differential diagnosis malignant from benign lesions and the presence tumor invasion in breast carcinomas. PMID- 11027872 TI - Long-term follow-up of 82 patients with chronic disease of the thoracic aorta using spin-echo and cine gradient magnetic resonance imaging. AB - The objective of this study to examine the clinical impact of magnetic resonance imaging in long-term follow-up of patients (pts) with chronic disease of the thoracic aorta such as coarctation of the aorta, chronic aortic dissection and true aortic aneurysm. A total of 322 magnetic resonance examinations obtained in 82 pts with chronic disease of the thoracic aorta (31 pts with coarctation of the aorta (CoA), 29 pts with chronic aortic dissection and 22 pts with true aneurysm) over a period of 0.25 to 13.5 (mean +/- SD: 6.5 +/- 3.4) years were retrospectively reviewed. Diameters of the thoracic aorta were measured at predefined levels and morphological and functional parameters of special interest were analysed in each patient group. Pts were classified as having constant or progressive disease and clinical end-points were defined as (re-)operation or death. 43 pts (52%) (CoA 15 pts, chronic dissection 16 pts, true aneurysm 12 pts) had constant findings. None of them underwent (re-)operation and seven patients (16%) died, three of them from their aortic disease more than five years later after their last magnetic resonance examination, one from an arrhythmogenic event, and in the remaining 3 pts the cause of death could not be definitely established. 39 pts (48%) (CoA 16 pts, chronic dissection 13 pts, true aneurysm 10 pts) had progressive disease as demonstrated by repetitive magnetic resonance imaging. Of these 39 pts 24 pts underwent (re-)operation, in 15 pts operation was postponed. Four pts died from their aortic disease. Repetitive magnetic resonance imaging is a clinically feasible technique for long-term follow-up of pts with chronic disease of the thoracic aorta because it can detect progressive disease in a large subset of pts requiring elective surgery. The results of magnetic resonance imaging provided the rationale for either (re-)operation or conservative management, thus guiding patient management. PMID- 11027874 TI - Fast adipose tissue (FAT) assessment by MRI. AB - We report a method of fast adipose tissue (FAT) assessment to characterize the quantity, and distribution of abdominal adipose tissue. Whole-volume coverage of the abdomen was obtained using 31 contiguous transverse T(1)-weighted images from 16 obese females. A radiologist manually traced all adipose tissue volumes in the images, while a physiologist used an automated method to measure adipose tissue in a single image at the level of the umbilicus. Automated analysis of the umbilicus-level image was significantly correlated with values obtained by manual analysis of the entire abdomen (p < 0. 001). There was good agreement between the automated umbilicus-level image method and the manual whole abdomen method for subcutaneous adipose tissue (r(2) = 0.958), visceral adipose tissue (r(2) = 0. 753), and total adipose tissue (r(2) = 0.941). The automated method required 6 min vs 2 h for the manual method. PMID- 11027876 TI - Spatial resolution in echo planar imaging: shifting the acquisition window in k space. AB - Single-shot echo planar imaging (EPI) is one of the most suitable techniques for very fast image acquisition, especially in functional MRI. In standard EPI schemes the k-space center is sampled in the middle of the acquisition train. This leads to longer echo times for higher spatial resolutions, as well as reduced signal intensity and signal-to-noise ratio. Therefore, echo shifting to lower echo times is often used. After a brief overview on the theoretical background of various point-spread-functions (PSF) computational simulations are presented, which quantify the modulation amplitude of a binary test object sampled with either standard, zero-filled or shifted k-space acquisition. The results suggest that echo-shifting with zero-filling is not advantageous, not even with small matrix sizes, and that echo-shifting with additional acquisition of outer k-space lines decreases the modulation amplitude only slightly. Simulations were also performed on noise-corrupted test objects, indicating that the use of the echo-shifted scheme causes resolution loss of up to 30% compared to the standard scheme for a 128 by 128 pixel matrix at a noise level of 20%. Finally, in vivo experiments using different echo shifts are presented and the characteristics of signal and noise with varying TE are quantified. PMID- 11027875 TI - Diagnosis of vertebral metastasis, epidural metastasis, and malignant spinal cord compression: are T(1)-weighted sagittal images sufficient? AB - The objective of this study was to determine whether T(1)-weighted sagittal images alone are adequate in the diagnosis of vertebral metastasis, epidural metastasis, and malignant spinal cord compression. Ninety-four complete magnetic resonance (MR) studies of the spinal column (a complete study consisting of T(1) weighted sagittal images, T(2)-weighted sagittal images, and T(1)- and/or T(2) weighted axial images) and 94 T(1)-weighted sagittal images alone (a subset of the complete studies) from 57 consecutive cancer patients over the last 2 years with clinically suspected cord compression were blindly and independently evaluated by four radiologists. The complete MR studies were used as the standard. Overall, the sensitivity of T(1)-weighted sagittal images alone to vertebral metastasis (87%) was statistically greater than cord compression (70%) (p = 0.05), and statistically greater than epidural metastasis (46%) (p or= 75%) reduction in injection (ARR = 2.85, 95% confidence limit [CL] 1.47-5.51), to stop injecting altogether (ARR = 3.5, 95% CL 2.1-5.9), and to remain in drug treatment. New users of the exchange were five times more likely to enter drug treatment than never-exchangers. We conclude that reduced drug use and increased drug treatment enrollment associated with needle exchange participation may have many public health benefits, including prevention of blood borne viral transmission. PMID- 11027895 TI - Impulsivity and voucher versus money preference in polydrug-dependent participants enrolled in a contingency-management-based substance abuse treatment program. AB - Thirty-four polydrug-dependent participants enrolled in a voucher-based substance abuse treatment program were given choices between hypothetical amounts of money and hypothetical amounts of vouchers, which are traded for goods and services, to determine their preferences for the two payment modalities. It was hypothesized that the majority of participants would prefer money to voucher because under the circumstances of the treatment program, the delay associated with money exchange is shorter than the delay associated with voucher exchange. It was further hypothesized that those participants who selected money over voucher also would have greater levels of impulsivity as assessed by the Barratt Impulsiveness Rating Scale (BIS) (Barratt, 1965). The results show large individual differences in money/voucher preference with approximately half of the participants preferring money to voucher when the two amounts are equivalent. In addition, as the magnitude of the money/voucher comparisons increased from 0.50 dollars to 32.00 dollars, the percentage of participants that preferred money increased. No correlations were found between money/voucher preference and impulsivity scores. PMID- 11027896 TI - Depression after alcohol treatment as a risk factor for relapse among male veterans. AB - We examined the association between relapse-to-drinking and depressive symptomatology measured during inpatient treatment for alcohol disorder and 3 months posttreatment. Data were obtained from 298 veterans who completed 21-day inpatient treatment. Follow-up interviews were conducted at 3, 6, 9, and 12 months posttreatment. We used multiple logistic regression to assess the association between relapse and baseline/3-month posttreatment measures of depression (Beck Depression Inventory; BDI), controlling for important covariates. Our results showed that (a) the mild-to-moderately symptomatic participants (BDI = 14-19) at 3 months posttreatment were on average 2.9 times more likely than the nondepressed to have relapsed across follow-ups, and (b) the severely symptomatic participants (BDI = 20+) at 3 months posttreatment were on average 4.9 times more likely to have relapsed across follow-ups. Other analyses revealed that those with persistent depressive symptomatology reported at both baseline and 3 months posttreatment did not experience worse outcomes that those who reported symptomatology at 3 months posttreatment alone. PMID- 11027897 TI - Measuring client clinical progress in therapeutic community treatment. The therapeutic community Client Assessment Inventory, Client Assessment Summary, and Staff Assessment Summary. AB - The therapeutic community (TC) Client Assessment Inventory (CAI), Client Assessment Summary (CAS), and Staff Assessment Summary (SAS) are instruments developed from a comprehensive theory of TC treatment and recovery. They measure client self-report and staff evaluation of client progress along 14 domains of behavior, attitude, and cognitive change. The present article reports on the development of the instruments and findings from an analysis of data on 346 clients in TC treatment; including scale properties and cross-sectional client differences by treatment tenure. The instruments were administered over a 1-year period at two residential TC facilities. Findings indicate that both client and staff instruments reliably differentiate client clinical changes during treatment and client self-ratings are initially consistently higher than staff ratings. Clarifying and measuring changes in the individual during treatment is the first step in the effort to understand the change process and what steps can be taken to improve treatment effectiveness. PMID- 11027898 TI - Perceptions of staff attributes in substance abuse treatment. AB - Qualified professional staff contribute significantly to successful health-care service delivery. Organizations view six categories of staff attributes as valued qualities of competent personnel: knowledge and experience, organizational citizenship, interpersonal skills, service orientation, personal attributes, and leadership skills. This study presents the perceptions regarding these and other staff attributes held by female clients and staff from a substance abuse treatment facility. Results indicated that four attributes were perceived as particularly influential in assisting women to reach treatment goals. These included knowledge and experience, supportiveness, nonthreatening behaviors, and availability. Attention to these variables may prove useful as treatment programs strive to improve client outcomes. PMID- 11027899 TI - Mental health versus substance abuse treatment programs for dually diagnosed patients. AB - The aim of this study was to assess the similarities and differences of patients with co-existing psychiatric and substance use disorders attending treatment in either a mental health setting or a substance abuse treatment setting. A total of 129 patients were assessed, including 65 individuals from the substance abuse treatment center and 64 individuals from the mental health program. Treatment records were reviewed for diagnoses and sociodemographic data. While the two groups were highly similar with regard to age and ethnicity, there were significant differences in psychiatric profile, with the substance abuse treatment group having less severe diagnoses and no patients with schizophrenia, while the mental health treatment group had a majority of patients with schizophrenia. Other differences in the two groups, such as marital and parental status, disability status, and medical problems appeared to be directly linked with the aforementioned diagnostic profile. These data suggest important differences in characteristics of patients with comorbid disorders that appear to be dependent on the type of treatment program they attend. For the most effective management, integrated treatment programs should be aware of these differences and tailor service provision accordingly. PMID- 11027900 TI - Building culturally sensitive substance use prevention and treatment programs for transgendered populations. AB - Studies show that transgendered individuals are at high risk for substance use problems. It is important to identify the unique needs and concerns of these individuals and culturally sensitive programs that will be successful in recruiting and retaining these individuals in drug abuse treatment services. This involves incorporating the needs of services from the perspectives of both the transgendered community and health-care professionals. It is the purpose of this article to discuss transgenderism as well as the substance use problems and difficulties within substance use treatment that transgendered men and women may face. This article presents guidelines for the design and evaluation of health care services to transgendered populations. PMID- 11027901 TI - Methadone detoxification of tramadol dependence. AB - Tramadol hydrochloride is a centrally acting analgesic with a partial affinity for the opiate receptor (mu), having an analgesic potency estimated to be one tenth that of morphine. While preclinical investigations suggested that abuse liability associated with tramadol use is low, there are increasing numbers of cases reported to the U.S. Food and Drug Administration of abuse, dependence, and withdrawal associated with tramadol use. A case of a patient with tramadol dependence requiring detoxification with methadone is presented. Acute management of significant tramadol dependence has not yet been reported in the literature. Long-term treatment issues are also discussed. PMID- 11027902 TI - Dihydrocodeine: a useful tool in the detoxification of methadone-maintained patients. AB - We investigated the merit of dihydrocodeine tartrate for withdrawal in detoxifying 20 methadone-maintained patients presenting for treatment at the Leeds Addiction Unit (LAU). Thirteen patients (65%) successfully completed methadone detoxification and were abstinent from both methadone and opiate-type drugs at the end of the 2-week program. On completion, three patients began treatment with Naltrexone and another was abstinent at a follow-up appointment, 1 week later. A further patient relapsed back to heroin use but remained in contact with the LAU. The remaining six patients dropped out of the 2-week detoxification program between days 3 and 11 of the dihydrocodeine cross-over period. We believe dihydrocodeine may have advantages in detoxifying methadone-maintained patients. PMID- 11027903 TI - Gender-role stereotypes and interpersonal behavior how addicted inpatients view their ideal male and female therapist. AB - This study focuses on the influences of self-perceived interpersonal behavior of addicted inpatients (n = 107) on the stereotypes of their ideal male and female therapist. Based on the interpersonal model of personality patients were asked to describe their ideal male and female therapist. Perceptions of interpersonal behavior of the ideal therapist are determined by the self-perceived interpersonal behavior of the patients rather than by their gender. Ideal therapists are described as managerial-autocratic and sociable-extravert, which is in almost perfect accordance with the self-perceptions of therapists. Although most of the patients prefer a female therapist, the interpersonal behavior of a male or a female therapist does not differ significantly. The results are discussed from the viewpoint that mutual expectations and preferences between patients and therapists are thought to be of crucial importance for the outcome of therapy. In the process of matching patients and therapists, these expectations should not only be assessed, but also used to improve the final matching. PMID- 11027904 TI - Pain responses in methadone-maintained opioid abusers. AB - Providing pain management for known opioid abusers is a challenging clinical task, in part because little is known about their pain experience and analgesic requirements. This study was designed to describe pain tolerance and analgesic response in a sample of opioid addicts stabilized in methadone-maintenance (MM) treatment (n = 60) in comparison to matched nondependent control subjects (n = 60). By using a placebo-controlled, two-way factorial design, tolerance to cold pressor (CP) pain was examined, both before and after oral administration of therapeutic doses of common opioid (hydromorphone 2 mg) and nonsteroidal anti inflammatory (ketorolac 10 mg) analgesic agents. Results showed that MM individuals were significantly less tolerant of CP pain than control subjects, replicating previous work. Analgesic effects were significant neither for medication nor group. These data indicate that MM opioid abusers represent a pain intolerant subset of clinical patients. Their complaints of pain should be evaluated seriously and managed aggressively. PMID- 11027905 TI - Analgesic effect of intravenous ketamine in cancer patients on morphine therapy: a randomized, controlled, double-blind, crossover, double-dose study. AB - Pain not responsive to morphine is often problematic. Animal and clinical studies have suggested that N-methyl-D-aspartate (NMDA) antagonists, such as ketamine, may be effective in improving opioid analgesia in difficult pain syndromes, such as neuropathic pain. A slow bolus of subhypnotic doses of ketamine (0.25 mg/kg or 0.50 mg/kg) was given to 10 cancer patients whose pain was unrelieved by morphine in a randomized, double-blind, crossover, double-dose study. Pain intensity on a 0 to 10 numerical scale; nausea and vomiting, drowsiness, confusion, and dry mouth, using a scale from 0 to 3 (not at all, slight, a lot, awful); Mini-Mental State Examination (MMSE) (0-30); and arterial pressure were recorded before administration of drugs (T0) and after 30 minutes (T30), 60 minutes (T60), 120 minutes (T120), and 180 minutes (T180). Ketamine, but not saline solution, significantly reduced the pain intensity in almost all the patients at both doses. This effect was more relevant in patients treated with higher doses. Hallucinations occurred in 4 patients, and an unpleasant sensation ("empty head") was also reported by 2 patients. These episodes reversed after the administration of diazepam 1 mg intravenously. Significant increases in drowsiness were reported in patients treated with ketamine in both groups and were more marked with ketamine 0.50 mg/kg. A significant difference in MMSE was observed at T30 in patients who received 0.50 mg/kg of ketamine. Ketamine can improve morphine analgesia in difficult pain syndromes, such as neuropathic pain. However, the occurrence of central adverse effects should be taken into account, especially when using higher doses. This observation should be tested in studies of prolonged ketamine administration. PMID- 11027906 TI - An assessment of the safety, efficacy, and acceptability of intranasal fentanyl citrate in the management of cancer-related breakthrough pain: a pilot study. AB - The effects of intranasal fentanyl citrate (INFC) were assessed in 12 hospice inpatients with cancer-related breakthrough pain. Patients received 20 microg of fentanyl citrate and were asked to rate their pain using a visual analogue scale (VAS) before INFC, then after 3, 5, 10, 15, 30, 45, and 60 minutes. Eight patients (66%) had reductions in pain scores, four within 5 minutes and seven within 10 minutes of taking INFC. Ratings for INFC were very good (5 = 42%), good (3 = 25%), moderate (1 = 8%), and bad (3 = 25%). In comparison to oral morphine, INFC was better (6 = 50%), the same (3 = 25%), or worse (3 = 25%). Nine patients (75%) said they would continue to use INFC. Of the three patients who did not experience a positive result, two were taking relatively higher baseline opioid doses and one was found to have a fracture. No systemic adverse events were noted; two patients reported nasal itching or discomfort on first use that disappeared with repeated use. Intranasal fentanyl citrate appears safe and well tolerated by these patients. Randomized placebo-controlled and dose-ranging studies are required to confirm these findings. PMID- 11027907 TI - Criterion validation of the Edinburgh postnatal depression scale as a screening tool for depression in patients with advanced metastatic cancer. AB - It is estimated that 25% of palliative care patients will have identifiable symptoms of depression. Near the end of life, the distinction between what can be called "appropriate sadness" and depression may be difficult. Many palliative care units use rating scales to help identify patients who may be depressed. It is believed that symptoms such as guilt, worthlessness, and hopelessness may be more discriminating than other symptoms for depression within this population. The Edinburgh postnatal depression scale (EPDS) was devised for use in women in the postnatal period and does not contain any somatic-type symptoms. It consists of 10 items, each rated on a four-point scale, and includes items on guilt, thoughts of self-harm, and hopelessness. It has not previously been used for screening in cancer patients. In a study of 100 inpatients receiving palliative care, a cutoff of 13 on the EPDS had a sensitivity of 81% and a specificity of 79% for detecting cases of depression. There was a low rate of misclassifications. This study suggests that the EPDS may be useful as a screening instrument for palliative care patients. PMID- 11027908 TI - Avoidance of colostomy placement in advanced colorectal cancer with twice weekly hypofractionated radiation plus continuous infusion 5-fluorouracil. AB - The purpose of this study was to determine the efficacy of twice weekly hypo fractionated radiation therapy (RT) plus continuous infusion 5-fluorouracil for unresectable or locally advanced colorectal cancer with synchronous metastases. Palliative radiation consisting of 30 Gy/6 fractions/3 weeks was administered to 87 patients from 1982-1995 with 3 field belly board technique. Diverting colostomy was required for obstruction in 14 (16%) prior to radiation; 47 patients (54%) had recurrent disease following prior resection and 58 (66%) had distant metastases on presentation. Median follow-up was 12 months (1-104 months). Stabilization/regression of pelvic disease was accomplished in 65 patients (75%). Complete resection of the pelvic disease was accomplished in 5 patients (6%), all of whom had synchronous metastases. Diverting colostomy was required in only 11 patients after chemoradiation because of progressive pelvic tumor; 2 of these 11 patients (18%) had isolated progression of pelvic disease. Overall, colostomy was not required in 48 of the 72 patients (67%) who did not present with a stoma. Median survival was 11 months if metastatic disease was present and only 6 months when disease was limited to the pelvis. Grade 3 acute radiation toxicities occurred in <10% and no grade 4 acute toxicities were observed. No significant late radiation effects were noted. Twice weekly hypofractionated RT is well tolerated and provides durable palliation of symptoms related to locally advanced primary or recurrent colorectal cancer with metastatic disease. PMID- 11027909 TI - Manual acupuncture reduces hyperemesis gravidarum: a placebo-controlled, randomized, single-blind, crossover study. AB - Hyperemesis gravidarum, severe vomiting, develops in about 1-2% of all pregnancies. Acupuncture on the point PC6 above the wrist on the palmar side has been found to prevent some types of nausea and vomiting. The purpose of the present study was to see if acupuncture, in addition to standard treatment, could hasten the improvement of hyperemesis gravidarum. Thirty-three women with hyperemesis were evaluated in a randomized, single-blind, crossover comparison of two methods of acupuncture, active (deep) PC6 acupuncture or placebo (superficial) acupuncture. The women estimated their degree of nausea on a visual analogue scale (VAS). The daily number of emesis episodes were documented. Crossover analyses showed that there was a significantly faster reduction of nausea VAS and more women who stopped vomiting after active acupuncture than after placebo acupuncture. This study suggests that active PC6 acupuncture, in combination with standard treatment, could make women with hyperemesis gravidarum better faster than placebo acupuncture. PMID- 11027910 TI - Gabapentin vs. amitriptyline in painful diabetic neuropathy: an open-label pilot study. AB - The objective of this study was to compare the efficacy and tolerability of gabapentin and amitriptyline monotherapy in painful diabetic neuropathy. This was a 12-week, open-label, prospective, randomized trial. Twenty-five type-II diabetic patients with pain attributed to diabetic neuropathy and a minimum score of 2 on a pain intensity scale ranging from 0 (no pain) to 4 (excruciating pain) were randomized to receive either gabapentin, titrated from 1,200 mg/day to a maximum of 2,400 mg/day, or amitriptyline, titrated from 30 mg/day to a maximum of 90 mg/day. Both drugs were titrated over a 4-week period and maintained at the maximum tolerated dose for 8 weeks. The main outcome measures were weekly pain intensity and paresthesia intensity, measured on two categorical scales. Thirteen patients received gabapentin and 12 received amitriptyline. All 25 patients completed the trial. Gabapentin produced greater pain reductions than amitriptyline (mean final scores were 1.9 vs. 1.3 points below baseline scores; P = 0.026). Decreases in paresthesia scores also were in favor of gabapentin (1.8 vs. 0.9 points; P = 0. 004). Adverse events were more frequent in the amitriptyline group than in the gabapentin group: they were reported by 11/12 (92%) and 4/13 (31%) of patients, respectively (P = 0.003). Side effects were the main limiting factor preventing dose escalation. Gabapentin produced greater improvements than amitriptyline in pain and paresthesia associated with diabetic neuropathy. Additionally, gabapentin was better tolerated than amitriptyline. Further controlled trials are needed to confirm these preliminary results. PMID- 11027911 TI - Course of symptoms and quality of life measurement in Complex Regional Pain Syndrome: a pilot survey. AB - Few data have been published regarding the natural history, course of symptoms, and quality of life in Complex Regional Pain Syndrome (CRPS). To obtain preliminary data regarding these important issues in CRPS, a set of patient self report questionnaires were mailed to patients with the diagnosis of CRPS who had been assessed and/or treated at a tertiary university-based pain center in the United States. Self-reports of demographic information, symptoms, the Neuropathic Pain Scale, and a modified Brief Pain Inventory (mBPI) were received from 31 CRPS patients. Approximately 75% of patients reported initial symptoms of pain, swelling, coldness, and color changes. An additional 71% had weakness and inability to move the extremity as initial symptoms. Weakness at some time during their course of CRPS was described by 97%. A majority reported no overall improvement or worsening of symptoms over time (mean 3.3 years). The pain descriptors with the highest mean values were "deep" (6.4/10), "unpleasant" (6.4), "sensitive" (5.7), "surface" (5.4), and "dull" (5.3) pains. Significant sleep disturbance was reported by 80%. CRPS had a severe impact on quality of life, with substantial interference reported in 9 of 10 mBPI activity items by a majority of these patients. These findings should be viewed with caution and should not be generalized to the entire CRPS population because the cohort was small and select. A large multicenter prospective study needs to be performed to validate these preliminary findings. PMID- 11027912 TI - Adherence monitoring and drug surveillance in chronic opioid therapy. AB - Monitoring adherence with chronic opioid therapies is a critical yet often difficult task. Because chronic opioid therapy is often fraught with complex pharmacological, psychological, social, and legal issues, its application is often controversial or altogether avoided. Improved drug monitoring and surveillance may help reduce some of the reluctance to use chronic opioid therapy in patients with chronic pain states. We review the literature on patient adherence/compliance with chronic administration of opioids as well as novel methods by which adherence with opioid therapy can be measured. PMID- 11027913 TI - Amputation and the prevention of phantom pain. AB - Although it has been proposed that preoperative analgesia with epidural administration of analgesics may prevent long-term phantom pain, published results to date have been contradictory and controversial. In this case report, we describe a 41-year-old man with local recurrence of squamous cell carcinoma of the anus who underwent a hemipelvectomy. Preoperatively he had a significant neuropathic pain syndrome requiring oxycodone 60 mg every 4 hours. An epidural infusion of morphine and bupivacaine was started 24 hours preoperatively and discontinued on the third postoperative day. Over the next 10 days the oxycodone was gradually decreased and eventually discontinued prior to discharge. A review of the literature reveals conflicting reports on the benefit of preoperative epidural pain management in the prevention of postoperative pain syndromes. Conflicting research and conclusions of commentators leaves unanswered questions for clinicians. Nevertheless, we do know that we need to provide the best pain relief for patients both before and after amputation. This may require a combination of the oral, subcutaneous or intravenous, and epidural routes. PMID- 11027914 TI - The corticosteroid receptor hypothesis of depression. AB - Signs and symptoms that are characteristic for depression include changes in the setpoint of the hypothalamic-pituitary-adrenocortical (HPA) system, which in the majority of these patients result in altered regulation of corticotropin (ACTH) and cortisol secretory activity. More refined analysis of the HPA system revealed that corticosteroid receptor (CR) signaling is impaired in major depression, resulting among other changes, in increased production and secretion of corticotropin-releasing hormone (CRH, also frequently abbreviated CRF) in various brain regions postulated to be involved in the causality of depression. This article summarizes the clinical and preclinical data, supporting the concept that impaired CR signaling is a key mechanism in the pathogenesis of depression. Mouse genetics, allowing for selective inactivation of genes relevant for HPA regulation and molecular pharmacology, dissecting the intracellular cascade of CR signaling, are the most promising future research fields, suited for identifying genes predisposing to depression. Focusing on these two research lines may also allow to gain insight into understanding how current antidepressants work and further, how more specific targets for future antidepressant drugs can be identified. PMID- 11027915 TI - Hormonal correlates of hand preference in free-ranging primates. AB - In this research we examined hormonal correlates of hand preference in free ranging primates. Specifically, we tested the hypothesis that levels of the stress hormone cortisol and the male sex hormone testosterone are correlated with handedness in male rhesus macaques (Macaca mulatta). We found significant positive relationships between cortisol and testosterone levels sampled during adolescence and the frequency of right- versus left-hand use sampled during adulthood. These data indicate that adolescent measures of cortisol and testosterone are correlated with hemispheric specialization in adult free-ranging primates. PMID- 11027916 TI - Limbic activation and psychophysiologic responses to aversive visual stimuli. Interaction with cognitive task. AB - We mapped regional brain activity and peripheral psychophysiologic responses, occurring in response to evocative emotional stimuli, and examined whether task instructions could modulate limbic activation. Ten subjects viewed pictures with neutral or aversive emotional content during simultaneous measurement of peripheral psychophysiology and brain activity with [15O]water positron emission tomography (PET). Cognitive task was manipulated by having the subjects rate the pictures or perform a recognition memory task. Aversive pictures, relative to neutral pictures, increased cerebral activity in bilateral amygdala, thalamic/hypothalamic area, midbrain, and left lateral prefrontal cortex, along with greater skin conductance responses (SCR). Voxel-by-voxel correlation coefficients between regional brain activity and SCR showed significant positive correlation peaks in the thalamus and right amygdala. Limbic activation was significantly greater during the rating condition compared to the recognition condition, suggesting that when task demands modify emotional responses, this modulation can occur at the level of limbic activity. PMID- 11027917 TI - "Early" and "late" effects of sustained haloperidol on apomorphine- and phencyclidine-induced sensorimotor gating deficits. AB - Both dopamine (DA) agonists and NMDA antagonists produce prepulse inhibition (PPI) deficits in rats that model PPI deficits in schizophrenia patients. While DA agonist effects on PPI are reversed by acute treatment with either "typical" high-potency D2 DA antagonists or "atypical" antipsychotics, PPI deficits produced by phencyclidine (PCP) are preferentially reversed by acute treatment with "atypical" antipsychotics. Acute effects of antipsychotics may not accurately model the more clinically relevant effects of these drugs that emerge after several weeks of continuous treatment. In the present study, sustained treatment with haloperidol via subcutaneous minipumps blocked the PPI-disruptive effects of apomorphine and attenuated the PCP-induced disruption of PPI. Restoration of PPI in apomorphine-treated rats was evident within the first week of sustained haloperidol administration. A partial reversal of PCP effects on PPI did not develop until the second week of sustained haloperidol treatment, followed a fluctuating course, but remained significant into the seventh week of sustained haloperidol administration. The delayed emergence of anti-PCP effects of haloperidol suggests that the brain substrates responsible for the DAergic and NMDA regulation of PPI are differentially sensitive to acute and chronic effects of antipsychotics. PMID- 11027918 TI - Sulpiride, a D2/D3 blocker, reduces cataplexy but not REM sleep in canine narcolepsy. AB - Cataplexy, an abnormal manifestation of REM sleep atonia, is currently treated with antidepressants. These medications also reduce physiological REM sleep and induce nocturnal sleep disturbances. Because a recent work on canine narcolepsy suggests that the mechanisms for triggering cataplexy are different from those for REM sleep, we hypothesized that compounds which act specifically on cataplexy, but not on REM sleep, could be developed. Canine studies also suggest that the dopamine D2/D3 receptor mechanism is specifically involved in the regulation of cataplexy, but little evidence suggests that this mechanism is important for REM sleep regulation. We therefore assessed the effects of sulpiride, a commonly used D2/D3 antagonist, on cataplexy and sleep in narcoleptic canines to explore the possible clinical application of D2/D3 antagonists for the treatment of human narcolepsy. Both acute and chronic oral administration of sulpiride (300 mg/dog, 600 mg/dog) significantly reduced cataplexy without noticeable side effects. Interestingly, the anticataplectic dose of sulpiride did not significantly reduce the amount of REM sleep. Sulpiride (and other D2/D3 antagonists) may therefore be an attractive new therapeutic indication in human narcolepsy. PMID- 11027919 TI - Regulation of ionotropic glutamate receptor subunits in different rat brain areas by a preferential sigma(1) receptor ligand and potential atypical antipsychotic. AB - The effect of chronic administration of the putative atypical antipsychotic E 5842, a preferential sigma(1) receptor ligand, on ionotropic glutamate receptor subunit levels of mRNA and protein, was studied. The repeated administration of E 5842 differentially regulated levels of the NMDA-2A and of GluR2 subunits in a regionally specific way. Levels of immunoreactivity for the NMDA-2A subunit were up-regulated in the medial prefrontal cortex, the frontoparietal cortex, the cingulate cortex, and in the dorsal striatum, while they were down-regulated in the nucleus accumbens. Levels of the GluR2 subunit of the AMPA receptor were up regulated in the medial prefrontal cortex and the nucleus accumbens and down regulation was observed in the dorso-lateral striatum. Regulation of the levels of mRNA for the different subunits was also observed in some cases. The results show that E-5842, through a mechanism still unknown, is able to modify levels of several glutamate receptor subunits and these changes could be related to its antipsychotic activity in pre-clinical tests. PMID- 11027920 TI - WAY 100135, an antagonist of 5-HT1A serotonin receptors, attenuates psychotomimetic effects of MK-801. AB - In the present study, we investigated whether the antagonist of 5-HT1A receptors, WAY 100135, was capable of modifying the psychostimulant and psychotomimetic effects of MK-801, a non-competitive antagonist of NMDA receptors. It was found that: 1) WAY 100135 (10 and 20 mg/kg, but not 1.25, 2.5, and 5 mg/kg) transiently, in a dose dependent manner, attenuated the locomotor stimulant effects of MK-801 (0.4 mg/kg). Given alone, WAY 100135 had no effect on the locomotor activity of rats; 2) WAY 100135 (1.25 and 2.5 mg/kg, but not 10 or 20 mg/kg), attenuated or abolished the disruptive effects of MK-801 on the sensorimotor gating measured in a prepulse-induced inhibition of the acoustic startle response paradigm. WAY 100135 in all tested doses had no effect on the sensorimotor gating or amplitude of the acoustic startle response; 3) WAY 100135 (1.25, 2.5 mg/kg, but not 5 mg/kg) attenuated the detrimental effects of MK-801 on working memory and selective attention, measured in a delayed alternation task. Again, given alone, WAY 100135 did not influence the behavior of rats in that experimental paradigm; and 4) MK-801 (0.4 mg/kg) had no effect on the 5-HT1A receptor mRNA level in rat hippocampus, measured 2 and 24 hours after MK-801 administration. These data indicate that 5-HT1A receptors might be involved in the psychotomimetic effects of non-competitive NMDA receptor antagonists. In addition, 5-HT1A serotonin receptor antagonists and partial agonists may have potential antipsychotic properties. PMID- 11027921 TI - Intravenous cocaine decreases cardiac vagal tone, vagal index (derived in lorenz space), and heart period complexity (approximate entropy) in cocaine abusers. AB - We assessed the effects of i.v. cocaine on parasympathetic and sympathetic nervous system activity, and on the complexity vs. regularity of changes in heart rate over time. Fourteen otherwise healthy men with histories of i.v. cocaine abuse received bolus injections of cocaine (20 mg or 40 mg) and placebo (saline) on different days. Cardiovascular measures derived from the electrocardiogram, including heart rate, Porges' vagal tone (respiratory sinus arrhythmia), the 0.10 Hz rhythm, Toichi's vagal index, Toichi's sympathetic index, and approximate entropy (ApEn), were measured continuously. As predicted, cocaine produced tachycardia, accompanied by pronounced decreases in response to 40 mg cocaine in two different vagal tone indexes that precisely mirrored the increases in heart rate. The measure of sympathetic (and vagal) neural influences on the heart (0.10 Hz wave) also decreased in response to cocaine. Converging evidence from Toichi's vagal index supported the conclusion that the tachycardia from cocaine was due to withdrawal of cardiac vagal tone. These findings, and evidence that cocaine decreased cardiovascular complexity, contradict the prevailing assumption that the mechanism by which cocaine produces tachycardia is sympathetic (beta adrenergic). We discuss implications for cardiac arrhythmias associated with cocaine abuse and death due to overdose. PMID- 11027923 TI - Effects of the CRH receptor antagonist CP-154,526 on intravenous cocaine self administration in rats. AB - The role for corticotropin-releasing hormone (CRH) receptors in the maintenance of intravenous cocaine self-administration in rats was investigated using the centrally active, small molecule CRH1 receptor antagonist CP-154,526. In these experiments, adult male Wistar rats were allowed alternating 15-min periods of access to food reinforcement and cocaine self-administration (0.125, 0.25 or 0. 5 mg/kg/infusion) during daily 2-h sessions. A 1-min timeout separated access to the two reinforcers. Pretreatment with CP-154, 526 produced dose-related decreases in cocaine self-administration without affecting food-reinforced responding, suggesting a specific effect of the antagonist on cocaine-maintained behavior. Drug intake was decreased across several doses of cocaine, with the dose-response curve for cocaine self-administration shifted downward and flattened, suggesting that CP-154,526 decreased cocaine reinforcement. Furthermore, responding on the cocaine lever following CP-154,526 pretreatment was significantly suppressed, even during the first 15 min of the session, a time when rats typically sample the cocaine lever during extinction, suggesting that CRH receptors may also be involved in some of the conditioned effects of cocaine as well. These data are discussed in terms of the role for CRH in the neurobehavioral effects of cocaine. PMID- 11027922 TI - Behavioral evidence for interactions between a hallucinogenic drug and group II metabotropic glutamate receptors. AB - Recent electrophysiological studies in our laboratory have demonstrated a physiological interaction between 5-HT(2A) and metabotropic glutamate2/3 (mGlu2/3) receptors in the medial prefrontal cortex. Several behavioral studies have found that phenethylamine hallucinogens with partial agonist activity at 5 HT(2A) receptors induce head shakes when directly administered into the medial prefrontal cortex. The purpose of the present experiments was to examine whether an interaction occurs between mGlu2/3 and 5-HT(2A) receptors on a behavioral level using head shakes induced by phenethylamine hallucinogens as a model of 5 HT(2A) receptor activation. Administration of the mGlu2/3 agonist LY354740 (0.3 10 mg/kg, ip) suppressed head shakes induced by the phenethylamine hallucinogen 1 (2, 5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI). Conversely, administration of the mGlu2/3 antagonist LY341495 (1 mg/kg, ip) enhanced the frequency of DOI induced head shakes. Taken together, these results raise the possibility that the psychomimetic properties of hallucinogenic drugs may be mediated in part, via increased glutamate release following activation of 5-HT(2A) receptors. PMID- 11027925 TI - Clinician, parent, and child prediction of medication or placebo in double-blind depression study. AB - To evaluate how well a blind is maintained in a double-blind study. Clinicians (n = 66), parents (n = 62), and depressed child/adolescent subjects (n = 62) predicted whether the patient had been on either placebo or active medication at the end of an eight-week double-blind placebo versus fluoxetine trial. Clinician, patient and parents' guesses as to which treatment they had received were at a chance level based on an overall analysis. However, when clinical response and condition assignment were controlled, all were correctly predicting placebo treatment but not medication treatment. The finding that subjects, parents and clinicians predict at a chance level is important for double-blind study design integrity. However, clinicians, parents and subjects were accurately predicting placebo treatment when clinical response and the assigned condition were taken into account but not medication. Since they do not know condition however, all remain essentially blinded, and this is an important finding for design and analysis integrity for double-blind studies. PMID- 11027924 TI - Allelic variation in the serotonin transporter promoter affects onset of paroxetine treatment response in late-life depression. AB - The relationship of the serotonin transporter gene promoter region polymorphism (5-HTTLPR) to antidepressant response was examined in 95 elderly patients receiving a protocolized treatment for depression with paroxetine or nortriptyline. Patients were treated for up to 12 weeks and assessed weekly with clinical ratings and measurements of plasma drug concentrations. Twenty-one of the paroxetine-treated subjects were found to have the ll genotype and 30 had at least one s allele. There were no baseline differences between these groups in pretreatment Hamilton Rating Scale for Depression (HRSD) scores or anxiety symptoms. During acute treatment with paroxetine, mean reductions from baseline in HRSD were significantly more rapid for patients with the ll genotype than for those possessing an s allele, despite equivalent paroxetine concentrations. Onset of response to nortriptyline was not affected. Allelic variation of 5-HTTLPR may contribute to the variable initial response of patients treated with a selective serotonin reuptake inhibitor. PMID- 11027926 TI - Dopamine D(2) receptor blockade by haloperidol. (3)H-raclopride reveals much higher occupancy than EEDQ. AB - Two techniques are commonly used to measure antipsychotic induced dopamine D(2) occupancy in animals: competition with a reversible radioligand (3H-raclopride) or with an irreversible receptor inactivator (EEDQ). While both of these techniques have been used in the past, there is no direct and systematic comparison. In the first direct comparison of these two methods we find that the dose of haloperidol required for blocking 50% of the dopamine D(2) receptors was 0.02 mg/kg/sc (95% CI 0.018-0.022 mg/kg) as measured using 3H-raclopride method; but was significantly higher with the EEDQ method 0.14 mg/kg/s.c. (95% CI 0.048 0.224 mg/kg). The 3H-raclopride method showed significantly lesser variance (p = 0.02) despite the higher sensitivity. This seven-fold difference in the sensitivity of the two techniques to measure antipsychotic-induced D(2) occupancy explains discrepancies in the previous studies which have used these two methods and also suggest that for future studies the 3H-raclopride method is a more sensitive and, likely, a more valid reflector of true receptor occupancy. PMID- 11027927 TI - Erratum. 3, 4-methylenedioxymeth-amphetamine (MDMA) modulates cortical and limbic brain activity as measured by PMID- 11027928 TI - Reporting the study populations of clinical trials. Clear transmission or static on the line? AB - In contrast to attempts that have been made to measure the clarity of reporting of the methods of clinical trials in journal articles, we report here an attempt to measure the accuracy of methods reporting. We focus in this article on eligibility criteria as a test case for the reporting of clinical trial methods. We examined the reporting of eligibility criteria in the protocol, methods paper (if applicable), journal article, and Clinical Alert for articles appearing in print between January 1988 and September 1994 for which a Clinical Alert had been issued. Eligibility criteria were further classified into five categories in order to examine the content of information loss, if any. On average, 82% of protocol eligibility criteria were reported in methods papers. Journal articles and Clinical Alerts fared somewhat worse: 63% of criteria were reported in journal articles, 19% in Clinical Alerts. In all three categories of medical communication, the reporting of criteria that defined the study disease tended to be complete; reporting of criteria relating to trial precision, patient safety, legal and ethical concerns, and administrative considerations, was not complete. We found that criteria for clinical trial eligibility are frequently under reported in medical communications. Moreover, some of the criteria omitted are of considerable clinical importance. We suggest that in the design phase of clinical trials, proposed eligibility criteria be scrutinized closely. Those criteria that survive this scrutiny and that have clinical import must be reported upon fully and accurately when communicating trial results. PMID- 11027929 TI - A cognitive screening battery for dementia in the elderly. AB - The objective of this study is to propose a screening instrument for dementia based on a reduced number of neuropsychological tests. The sample consists in the pooled data of the five follow-up visits of the Paquid cohort study on cerebral aging: the estimation sample included 2792 subjects (8830 observations) and the validation sample included 985 subjects (2643 observations). Among scores significantly associated with dementia, we retained only those that increased the specificity of the model for a sensitivity of one. Seven neuropsychological tests and the MMSE subscores were considered. The most discriminant combination of tests included the MMSE and the subscores "orientation to time" and "recall three objects," the Benton Visual Retention Test, and Isaacs' Set Test of verbal fluency. The specificity of this screening instrument was 0. 77 for a sensitivity of 1. PMID- 11027930 TI - Associations between demographic and disease-related variables and disability over the first five years of inflammatory polyarthritis: a longitudinal analysis using generalized estimating equations. AB - OBJECTIVES: To investigate whether the relationship between demographic and disease-related variables and disability is constant during the first five years of inflammatory polyarthritis (IP) and to identify the contribution from involvement of specific joint areas to overall disability. METHODS: 684 patients referred to the Norfolk Arthritis Register were followed for five years using the Health Assessment Questionnaire (HAQ). The relationship between disability and demographic and clinical variables was analyzed using a multi-level modelling approach. RESULTS: Female gender, older age at symptom onset (> or = 64 years), joint involvement at six specific sites, joint tenderness and the number of deformed joints were all independently associated with disability (HAQ > or = 1.00). Similar results were obtained using a more stringent cut-off (HAQ > or = 1.50) or when analysis was restricted to the 325 patients who satisfied the 1987 ARA list criteria for rheumatoid arthritis. CONCLUSION: Disability, as measured by the HAQ, was associated with a large number of independent factors over the first five years of disease. PMID- 11027931 TI - Venous thrombosis in relation to fibrinogen and factor VII genes among African Americans. AB - We evaluated the relation between venous thrombosis and plasma fibrinogen levels, the HaeIII and BcI polymorphisms of the beta fibrinogen gene, and the MspI polymorphisms of the factor VII gene in a case-control study of African Americans. The study included 91 venous thrombosis cases and 185 control subjects obtained from a hospital in Atlanta, Georgia. High plasma fibrinogen was associated with increased risk of venous thrombosis, but the finding was not statistically significant. There was little association between the HaeIII polymorphisms and the BclI polymorphisms and the risk of venous thrombosis. The prevalence of the M2/M2 genotype of the factor VII gene was higher among cases than controls, but the difference was not statistically significant. The prevalence of the HaeIII H2 allele and the BclI B2 allele of the beta fibrinogen gene, both of which have been associated with slightly higher levels of plasma fibrinogen in most studies, is considerably lower among African-Americans in this study than it is among Whites in the United States and among Northern Europeans. The study is limited by its small size. However, despite this limitation, it supports the belief that increased plasma fibrinogen levels are associated with increased venous thrombosis risk. The study also indicated that the HaeIII and the BclI polymorphisms of the beta fibrinogen gene and the MspI polymorphisms of the factor VII gene are not strong determinants of venous thrombosis. PMID- 11027932 TI - Evaluating the administration of an office-based health survey in a primary care practice. AB - Self-administered waiting room questionnaires are popular tools for gathering health information from patients, but these data cannot be used for research purposes without confirming adequate sampling of the practice population and assessing the completeness and accuracy of patients' responses. Long-term data collection also requires avoiding an imposition on clinic operations. We developed a protocol to test these questions in a 9-week pilot study of 884 survey-eligible patients visiting a family practice clinic. We found an adequate proportion of eligible patients were approached (74%) and participated (89%), they provided relatively complete (82-98%) and accurate responses, and the impact on office operations was minimal (<2 min of staff time per participant). Some demographic differences in participation and survey item completion were identified. A systematic process for testing survey performance allowed us to not only document these findings, but also to rapidly identify problems and introduce solutions while the survey was in progress. PMID- 11027933 TI - Clinical features of tiaprofenic acid (surgam) associated cystitis and a study of risk factors for its development. AB - A case-control study was performed to describe the clinical course and identify risk factors predisposing to the development of tiaprofenic acid associated cystitis. Cases were identified from reports to the Australian Adverse Drug Reactions Advisory Committee and a two-year (1995-1996) national surveillance programme. Definition of a case was onset of symptoms of cystitis while taking tiaprofenic acid with pyuria (>10 WBC/microl) and/or hematuria (>10 RBC/microl), and no evidence of urinary tract infection. Definitions of "prescriber-matched" and "unmatched" controls were those who had been prescribed tiaprofenic acid by the same prescriber within 12 months of the case without developing cystitis or were identified from the databases of local pharmacies of the cases respectively. 81 of 109 identified cases and 109 of 184 potential controls completed a standardised interviewer-administered questionnaire. Median time between the commencement of tiaprofenic acid and symptom onset was 6. 3 months (range 0.1 47.1 months). Median interval between presentation to doctor and drug cessation was 3.0 months (range 0-24. 5 months). In half the patients, symptoms resolved within 14 days of ceasing therapy. Increasing age was a risk factor for the development of tiaprofenic-acid associated cystitis, age >70 years-odds ratio 3.2 (95% confidence interval 1.3-7.9) compared with age <55 years. Patients taking aspirin had a reduced risk (odds ratio 0.3, 95% confidence interval 0.1-0.9). Dose and amount of fluid consumed per day were not related to the risk of cystitis. Earlier recognition of tiaprofenic acid associated cystitis can potentially reduce the morbidity related to this condition. Apart from an increased risk among older patients, it is likely that this condition represents a drug reaction that cannot be predicted from clinical variables. PMID- 11027934 TI - Alcohol consumption had no beneficial effect on serum lipids in a substantial proportion of patients with primary hyperlipidemia. AB - The cardioprotective effect of moderate alcohol consumption, which is mediated largely by an elevation of high-density lipoprotein (HDL) levels, may be absent in patients with primary disorders of lipid metabolism. In 881 untreated, unrelated patients with primary hyperlipidemia, we estimated the effect of alcohol on fasting serum HDL cholesterol concentration adjusted for age, gender, diet, and smoking with a multiple linear regression model. The number of abstainers, moderate users (1-30 g daily), and heavy alcohol consumers (> or = 30 g daily) was not different between three types of hyperlipidemia. Among hypercholesterolemic (n = 514) and combined hyperlipidemic (n = 227) patients, consumers of alcohol had higher HDL levels compared to abstainers (P(for trend) = 0.0001 and 0.0002, respectively). Heavy drinkers with combined hyperlipidemia had higher triglyceride levels compared to moderate drinkers and abstainers (P(for trend) = 0.008). Alcohol consumption had no effect on the lipid profiles in 140 patients with hypertriglyceridemia. In conclusion, combined hyperlipidemic and hypertriglyceridemic patients may profit from an encouragement to stop consuming alcohol. On the other hand, moderate use of alcohol may delay the development of coronary artery disease in hypercholesterolemic patients by an elevation of HDL levels. PMID- 11027935 TI - Estimation of the amyotrophic lateral sclerosis incidence by capture-recapture method in the Limousin region of France. AB - Amyotrophic lateral sclerosis (ALS) is a rapidly fatal degenerative neurological disease categorized among motor neuron diseases. In the literature, the incidence of ALS varies between 0.7 and 2.4 per 100, 000 inhabitants. A study using the capture-recapture method (multiple records system analysis) was undertaken in Limousin (France) ascertaining all patients having onset of definite or probable ALS during the period 1994-1995. Three information sources able to identify these new ALS cases were selected: the first source was a computerized database of the Neurology Department of the University Hospital of Limoges; the second source consisted of the neurologists of the Limousin region and neighboring provinces (county-sized regions); the third source grouped the hospitals of the Limousin region and neighboring provinces (county-sized regions). During this period, 46 new cases of ALS were seen, corresponding to an observed mean annual incidence of 3.2 (+/-0.6) per 100,000 inhabitants. After standardization for age, the annual incidence was 2.5 per 100,000 inhabitants. The number of new cases estimated by the capture-recapture method was 70, corresponding to an estimated mean annual incidence of 4.9 (+/-1.0) for 100,000 inhabitants. Hence, statistical modeling utilizing partially overlapping information sources permitted a more exhaustive compendium of the new cases of ALS and may be a truer reflection of actual disease incidence than has been previously reported. PMID- 11027936 TI - Physician gender and cesarean sections. AB - BACKGROUND: Among consumers insurers, and providers there is pervasive concern regarding the high incidence of cesarean section delivery. To date, attempts to reduce these rates have focused on the clinical behavior of providers resulting in only minimal changes. Therefore, non-medical variables must be investigated as potential explanatory factors for the decision to perform cesarean delivery. METHODS: Data were collected on clinical and non-clinical factors for obstetrician-gynecologists delivering at Yale-New Haven Medical Center to measure the impact of these factors on the performance of cesarean sections. Specifically, variation in patient demographic, ante- and intra-partum risk variables, practice setting, and doctor-specific characteristics were examined. Using contingency table and logistic regression analyses the contribution of selected factors was evaluated. RESULTS: Multivariate modeling revealed that male physicians were significantly more likely than their female colleagues to perform cesarean section. This relationship was particularly strong in the university practice setting. CONCLUSIONS: Efforts to reduce the incidence of cesarean section need to focus on the continuing education of health care providers and the delineation of non-clinical factors as essential elements in the election of specific clinical therapies. PMID- 11027937 TI - Analyzing data with clumping at zero. An example demonstration. AB - This article demonstrates the use of two approaches to analyzing the relationship of multiple covariates to an outcome which has a high proportion of zero values. One approach is to categorize the continuous outcome (including the zero category) and then fit a proportional odds model. Another approach is to use logistic regression to model the probability of a zero response and ordinary least squares linear regression to model the non-zero continuous responses. The use of these two approaches was demonstrated using outcomes data on hours of care received from the Springfield Elder Project. A crude linear model including both zero and non-zero values was also used for comparison. We conclude that the choice of approaches for analysis depends on the data. If the proportional odds assumption is valid, then it appears to be the method of choice; otherwise, the combination of logistic regression and a linear model is preferable. PMID- 11027938 TI - Signs and symptoms of melanoma in older populations. AB - We conducted a descriptive study to assess the relationship between increasing age and the reporting of melanoma signs/symptoms in 634 hospital-based and 624 population-based incident cases of melanoma. Multivariate logistic regression was used to evaluate the relationship between older age (> or = 50 years) and the reporting of melanoma signs/symptoms. Older patients were less likely to report itching and change in elevation of their lesions (P < 0.05). Change in color was also less likely to be reported by older patients, although not statistically significant. Ulceration of the lesion was reported significantly more by older patients (P < 0.05). Older individuals may be less likely to report itching and change in elevation/color of their lesions, but more likely to report ulceration, a symptom associated with advanced disease and poor prognosis. Further research is necessary to provide a better understanding of the development of melanoma in older populations so that new strategies can be explored to improve early detection in this age group. PMID- 11027939 TI - Determinants of physician reluctance to perform mouth-to-mouth resuscitation. AB - OBJECTIVES: Mouth-to-mouth resuscitation (MMR) is widely taught and promoted. The purpose of this study was to better characterize the observation that health professionals are reluctant to perform MMR and to identify determinants of this reluctance. METHODS: 324 residents and faculty at a New York City teaching hospital were anonymously surveyed regarding their reluctance to perform MMR. One year later, medical staff were resurveyed. RESULTS: Reluctance varied across scenarios: 70-80% of physicians were willing to perform MMR on a newborn or child, 40-50% for an unknown man, and 20-30% for a trauma victim or potentially gay man. Physicians reported very similar percentages for each scenario in the two surveys. Factors associated with MMR reluctance were female gender (OR = 2), resident physician (OR = 2), and higher perceived risk of contracting HIV from MMR (OR = 1.4 per unit on 5-point scale). In the year before the survey, 30% of all respondents witnessed an apneic patient who required MMR for whom ventilation was not provided for at least 2 minutes. CONCLUSIONS: Many physicians are reluctant to perform MMR. Marked delays in ventilation of apneic patients are occurring. PMID- 11027940 TI - Mortality in a population with long-term exposure to inorganic selenium via drinking water. AB - We analyzed the 1986-1997 mortality in a cohort of 2065 residents of an Italian municipality which had been exposed to drinking water with a high content of inorganic selenium over a long period of time, and compared it with mortality in the remainder of the municipal population. Mortality from malignant neoplasms increased [standardized mortality ratio (SMR) 1.17, 95% confidence interval (CI) 0.96-1.42], mainly due to an excess mortality from melanoma and colorectal cancer in both sexes, kidney cancer in men, and lymphoid malignancies in women. Overall cardiovascular mortality changed little (SMR 1.05, 95% CI 0.89-1.23), despite the higher cerebrovascular mortality (SMR 1.43, 95% CI 1.03-1.93). Coronary disease mortality slightly decreased (SMR 0.87, 95% CI 0.63-1.16), due to a low mortality among women. We also noted an excess mortality from Parkinson's disease in men and from motor neuron disease in women. Evaluation of these findings is, however, hampered by the lack of information about potential lifestyle confounders, the fact that the exposure could only be characterized by a simple dichotomization, and the inconsistencies of most estimates between the two sexes. PMID- 11027941 TI - A new method for identifying antibiotic-treated infections using automated pharmacy records. AB - For research purposes, there are few alternatives to costly surveillance for ascertaining infections in community populations. We propose a new approach based on antibiotic prescription fills in automated pharmacy records of the Group Health Cooperative of Puget Sound, in Seattle, Washington, to identify treated infections in postmenopausal women. After excluding probable antimicrobial prophylaxis and chronic antibiotic use, four intervals between antibiotic fills (30, 45, 60, and 90 days) were tested for their ability to detect new infections. Concordance with outpatient medical record reviews was evaluated in 150 women. The sensitivity of the automated pharmacy records using the four cutpoints for detecting new infections ranged from 88 to 80%, from 30 to 90 days, respectively. Of the 81 women with no infection in the chart reviews, 75% also had no infection using the pharmacy method. Good agreement was found between the two methods for counts of infections per person over the 2-year follow-up, with the 60-day cutpoint showing the greatest overall agreement with chart reviews (kappa = 0.55). The pharmacy method presented here offers a useful new approach for infection ascertainment for epidemiologic research. PMID- 11027942 TI - Adverse drug reactions in children reported by means of the yellow card in Spain. AB - OBJECTIVE: To analyze the case reports concerning children (14 years or younger) in the Spanish Pharmacovigilance System over a 10-year period (1982-1991). FINDINGS: The study of 1419 reports of adverse drug reaction (9.8% of all those received) showed the most commonly involved organs and systems to be the skin, digestive tract, and nervous system (62.8%). The most commonly involved pharmacological groups were antibiotics, respiratory medications, and vaccines (69%). The absolute number of reports is higher in children between 1 and 4 years of age (37.9%). There were more reports among males than in females. Less than 5% of the reports notified directly life-threatening or fatal reactions. CONCLUSIONS: Adverse drug reaction are not common in pediatric patients, and most are mild. However, due to limitations of clinical trials in children, pharmacoepidemiological studies may be the only source of information on the benefit-risk profile of drugs received by these patients, and as such require special attention. PMID- 11027943 TI - beta-adrenergic mechanisms in cardiac diseases: a perspective. AB - Several lines of evidence show that neurohumoral systems, especially those involving catecholamines, play a crucial role in cardiac diseases. Changes in the beta-adrenergic receptor (beta-AR) system such as receptor down-regulation, uncoupling from G-proteins, receptor internalization and receptor degradation may account for some of the abnormalities of contractile function in this disease. Increases in the level of inhibitory G-protein subunits also appears to be involved in attenuating the beta-AR signal. Finally beta-AR signalling is strongly regulated by members of the G-protein-coupled receptor kinase family (GRKs), the best known of which is beta-adrenergic receptor kinase 1 (beta-ARK1). beta-ARK1 mRNA, protein level and enzymatic activity is increased in heart disease, further contributing to an attenuation in beta-AR signalling. The combination of these negative alterations are presumably related to the contractile dysfunction seen in human heart disease. The combination of biochemical, physiological and molecular biological studies bearing on the normal function and regulation of these various molecules should provide strategies for elucidating the pharmacological basis of the regulation of myocardial contractility in the normal and failing heart. PMID- 11027944 TI - Small G-protein networks: their crosstalk and signal cascades. AB - Small GTP-binding proteins (G-proteins) exist in eukaryotes from yeast to human and constitute a superfamily consisting of more than 100 members. This superfamily is structurally classified into at least five families: the Ras, Rho/Rac/Cdc42, Rab, Sar1/Arf, and Ran families. They play key roles not only in temporal but also in spatial determination of specific cell functions. It has become clear that multiple small G-proteins form signalling cascades that are involved in various cellular functions, such as budding processes of the yeast and regulation of the actin cytoskeleton in fibroblasts. In addition, two distinct small G-proteins regulate specific cellular functions in a cooperative or antagonistic manner. A single small G-protein exerts various biological responses through different downstream effectors. Moreover, some of these downstream effectors sequentially activate further downstream effector proteins. Thus, small G-proteins appear to exert their functions through their mutual crosstalk and multiple downstream effectors in a variety of cellular functions. PMID- 11027945 TI - STAT-signalling through the cytoplasmic compartment: consideration of a new paradigm. AB - The binding of a large number of cytokines and growth factors to their cognate receptors on the surface of mammalian-cell plasma membrane activates a signalling cascade involving the cytoplasmic STAT-family proteins, which is characterized by the nuclear translocation of a cytokine- or growth factor-specific subset of the cytoplasmic pool of the respective tyrosine- and serine-phosphorylated STAT proteins and the consequent transcriptional activation of specific target genes. In the standard model of cytokine-induced STAT signalling such as that elicited by various interferons and interleukins, it is thought that STAT proteins are recruited to the cytoplasmic side of the cell-surface receptor complex from within a monomeric cytosolic pool, and upon tyrosine-phosphorylation by respective Janus kinase family members, dimerize and translocate to the nucleus. The mechanisms which determine and regulate the recruitment of cytosolic STAT proteins to the plasma membrane-receptor complex, the transit of "activated" STATs through the expanse of the cytoplasmic compartment from the plasma membrane to the nuclear pore region, and the transit of STATs through the nuclear pore complex into the nuclear compartment, remain largely unknown. New data from different laboratories suggests consideration of a model for STAT signalling in which STAT proteins function in the cytoplasm not only as free monomers and dimers but as part of heteromeric complexes ("statosomes"), with accessory proteins which may serve to present specific STATs to the plasma membrane receptor complex, and to chaperone "activated" STATs through the cytoplasmic compartment toward the nucleus and then into the nuclear compartment. PMID- 11027946 TI - EGF triggers caveolin redistribution from the plasma membrane to the early/sorting endocytic compartment of hepatocytes. AB - In this study, we demonstrate that, in rat liver, epidermal growth factor (EGF) is responsible for the partial redistribution of caveolin-1 from the plasma membrane into the early/sorting endocytic compartment. Highly purified endosomes and plasma membrane fractions were isolated from control rat liver and from rats injected with EGF or pIgA for different times. Whereas in subcellular fractions from control hepatocytes most of caveolin was concentrated in the plasma membrane and the receptor-recycling fractions, after EGF injection there was a significant redistribution of caveolin toward the early/sorting (CURL) endocytic fractions. The recruitment of caveolin into the endocytic compartment was not induced by pIgA. PMID- 11027947 TI - The role of the cyclic GMP-inhibited cyclic AMP-specific phosphodiesterase (PDE3) in regulating clonal BRIN-BD11 insulin secreting cell survival. AB - We report here that the cyclic GMP-inhibited cyclic AMP specific phosphodiesterase (PDE3B) is expressed as a membrane-bound protein in clonal insulin-secreting BRIN-BD11 cells. This was shown using SKF94836 (PDE3 inhibitor) which maximally inhibited membrane-bound cyclic AMP PDE activity by approximately 25-30% and by RT-PCR. We also demonstrated that insulin growth factor-1 (IGF-1) activates PDE3B in BRIN-BD11 cells. We therefore evaluated the role of phosphoinositide 3-kinase (PI3K) and p42/p44 mitogen-activated protein kinase (p42/p44 MAPK) pathways in regulating this enzyme. We report here that the PI3K inhibitor, wortmannin, prevented the IGF-1-dependent stimulation of PDE3B activity. In contrast, the inhibitor of MEK-1 activation, PD098059 (which reduced IGF-1-stimulated p42/p44 MAPK phosphorylation), had no effect on PDE3B activation. Furthermore, IGF-1-dependent stimulation of p42/p44 MAPK and PDE3B was abolished in serum-deprived cells and this was associated with apoptosis. We propose that the deregulation of the PI3K/PDE3B pathway might result in increased intracellular cyclic AMP accumulation, which promotes apoptosis. This was supported by the finding that the adenylyl cyclase activator, forskolin, also induced apoptosis. Finally, we found that orthovanadate (a phosphotyrosine phosphatase inhibitor) fully restored the activation of p42/p44 MAPK in serum deprived cells, but had only a small effect on PDE activity. This confirmed that p42/p44 MAPK is on a separate pathway to PDE3B. Therefore, IGF-1-dependent regulation of PDE3B may be linked to cell survival through PI3K and not p42/p44 MAPK. PMID- 11027948 TI - Protein kinase C decreases the hepatocyte growth factor-induced activation of Erk1/Erk2 MAP kinases. AB - HGF and phorbol ester induce the scattering of HepG2 cells. Recently, we have reported that the motility and morphological responses that accompany this process require the activation of Erk1/Erk2 MAP kinases, and phosphatidylinositol 3-kinase contributes to the activation of Erk1/Erk2 in HGF-induced cells. The cell scattering-associated appearance of a high-M(r) (>300 kDa) protein pair has also been observed, and has been proven to be a sensitive marker of the intensity of Erk1/Erk2 activation. Our present study demonstrates that in HGF-induced cells protein kinase C and phosphatidylinositol 3-kinase regulate oppositely the expression of these cell scattering-associated proteins. While in phorbol ester treated cells the sustained activation of protein kinase C is essential for this expression, in HGF-induced cells the inhibition of protein kinase C with bisindolylmaleimide I stimulates the expression. Protein kinase C reduces the HGF induced phosphorylation of Erk1/Erk2, and in this way it can limit the intensity of Erk1/Erk2-dependent gene-expression PMID- 11027949 TI - cAMP-dependent protein kinase (PKA) inhibits T cell activation by phosphorylating ser-43 of raf-1 in the MAPK/ERK pathway. AB - cAMP-dependent protein kinase (PKA) has been suggested to interfere with T-cell activation by inhibiting interleukin (IL-2) receptor alpha-chain (CD25) expression and IL-2 production. The Ras/MAP kinase pathway has been found to be necessary for induction of the IL-2 production. In this study, we have scrutinized the Ras/MAP kinase pathway in Jurkat T-cells to attempt to identify any sites for PKA-mediated regulatory phosphorylations. Here we unambiguously demonstrate that PKA directly inhibits anti-CD3-induced MAP kinase activation. In vitro phosphorylation experiments showed that Raf-1 was extensively phosphorylated by PKA, while ERK2 and MEK were not. Phosphopeptide mapping identified Ser-43 of Raf-1 as the only site phosphorylated by PKA in the Ras/MAPK pathway. Transient transfection experiments demonstrated that mutations of Ser-43 of the Raf-1 kinase were rendered insensitive to cAMP-mediated inhibition. PMID- 11027950 TI - Difference between follistatin isoforms in the inhibition of activin signalling: activin neutralizing activity of follistatin isoforms is dependent on their affinity for activin. AB - We demonstrate the difference between the follistatin isoforms (FS-288 and FS 315), two activin-binding proteins, in the neutralizing activity for activin signalling. Transcriptional reporter assay using 3TP-Lux, an activin-responsive reporter construct, showed that the inhibitory effect of FS-288 on activin induced transcriptional response is more potent than that of FS-315. The potency was not influenced by the presence of heparan sulfates, by which FS, in particular FS-288, associates with cell surfaces at a high affinity. Furthermore, FS-288 inhibited the binding of activin to its type II receptor more markedly than did FS-315, as evidenced by surface plasmon resonance and affinity cross linking experiments. Moreover, the Kd of FS-288 and FS-315 for activin A was estimated to be 46.5+/-0.37 pM and 432+/-26 pM, respectively, by surface plasmon resonance experiments. These results indicate that the different potency between the two FS isoforms in the inhibition of activin activities depends on their affinity for activin A. PMID- 11027951 TI - Crosstalk between cytosolic pH and intracellular calcium in human lymphocytes: effect of 4-aminopyridin, ammoniun chloride and ionomycin. AB - Stimulation of lymphocytes by specific antigens is followed by the activation of different signal transduction mechanisms, such as alterations in the cytoplasmic levels of Ca(2+), H(+) and variations in membrane potential. To study interrelationships among these parameters, changes in pHi and Ca(2+) were measured with the fluorescent probes BCECF and Fura-2 in freshly isolated blood human lymphocytes. Moreover, membrane potential qualitative alterations were recorded with the fluorescent dye bis-oxonol. In a bicarbonate-free medium, cell alkalinization with NH(4)Cl slightly decreased intracellular Ca(2+) concentration ([Ca(2+)](i)) due to efflux of Ca(2+) from the cell. In contrast, an elevation of pHi induced with 4-AP increased [Ca(2+)](i), either in the presence or absence of external Ca(2+). The increase in Ca(2+)-free medium is likely to be due to Ca(2+) release from thapsigargin and caffeine-independent intracellular stores. Both 4 AP or NH(4)Cl induced a plasma membrane depolarisation, although with different kinetics. The ionosphere ionomycin increased pHi, Ca(2+) levels and also induced membrane depolarisation. Together, these observations demonstrate a lack of correlation between the magnitude of changes in pHi and Ca(2+). PMID- 11027952 TI - Small area variations in health related behaviours; do these depend on the behaviour itself, its measurement, or on personal characteristics? AB - In this paper we examine the patterning, by small areas, of four health related behaviours (smoking, alcohol consumption, diet, and exercise) in the West of Scotland, after controlling for a range of individual/household characteristics, using multilevel models. Smoking and drinking were measured both as binary and as continuous variables, and diet and exercise were each measured in two ways: 'good' (health promoting) and 'bad' (health damaging). 'Area effects' (unattributed variation by post code sector) were found for 'bad' diet only. 'Good' and 'bad' diet, 'bad' exercise patterns and current smoking were associated with postcode sector deprivation. For 'bad' diet this effect was found only for individuals in more affluent households, and for 'good' exercise and current smoking the association with area deprivation differed between adolescents and adults. We conclude that the influence of area on health related behaviours varies according to the behaviour and the way it is measured, and that the influence of area deprivation and/or of area can vary by age and household deprivation. PMID- 11027953 TI - Deprivation and poor health in rural areas: inequalities hidden by averages. AB - Poor health and social deprivation scores in 570 wards in East Anglia, UK, were much less associated in rural than in urban areas. The deprivation measure most closely related to poor health in the least accessible rural wards was male unemployment, but use of this measure did not remove the urban-rural gradient of association strength. Neither did replacing wards by smaller enumeration districts as the units of analysis. The differences between urban and rural correlations were removed by restricting the comparison to wards with the same unemployment range and combining pairs of rural wards with similar deprivation values. Apparent differences between rural and urban associations are therefore not due to the choice of deprivation indices or census areas but are artifacts of the greater internal variability, smaller average deprivation range and smaller population size of rural small areas. Deprived people with poor health in rural areas are hidden by favourable averages of health and deprivation measures and do not benefit from resource allocations based on area values. PMID- 11027954 TI - The stress process in neighborhood context. AB - The positive relation between socio-economic status (SES) and health, both mental and physical, is examined within a stress-process framework. Telephone survey data of adults age 45-74 are analyzed to test the roles of stressors and resources as mediators of the SES-health relation. Next, the stress process is tested in neighborhood context by splitting the sample in half according to residence in lower- or higher-SES neighborhoods. The relative impact of stressors on mental and physical health, and effectiveness of resources in protecting mental and physical health, are tested separately for both types of neighborhoods. The results indicate that social support is only protective of mental and physical health among residents of higher-SES neighborhoods. The implications of the results for future research are discussed. PMID- 11027955 TI - Factors associated with hysterectomy among women in Australia. AB - This study was to identify hysterectomy prevalence across urban, rural and remote areas of Australia and across states, to separate geographic variation from the effect of sociodemographic influences, and also to compare the quality of life of women who have and have not had hysterectomy. Data were collected from 14,072 women aged 45-50 years participating in the baseline survey of the Australian Longitudinal Study on Women's Health. The estimated prevalence of hysterectomy was 22%. Factors significantly associated with hysterectomy included living in a rural or remote area, state of residence, having private health insurance, lower levels of education, being married and having more than two children, having had other gynaecological and non-gynaecological surgical procedures, and more visits to general practitioners. Compared with women who had not had hysterectomy, women who had had hysterectomy had significantly poorer physical and mental health as measured by the SF-36 quality of life profile (adjusted mean PCS=45.7 vs 49.3, p<0.0001; adjusted mean MCS=46.9 vs 48.2, p<0.0001). PMID- 11027956 TI - A multilevel time-series model for the incidence of AIDS cases in Spain. AB - OBJECTIVE: The main aim of this research is to study the quantitative evolution of the incidence of AIDS in the 19 Spanish Communities. The hypothesis is that incidence follows a multilevel autoregressive model, where each Community shows random variability around a general process. METHOD: On the basis of official data on the number of existing AIDS cases, an autorregressive multilevel time series model was developed. RESULTS AND CONCLUSIONS: Analysis shows that the hypothesis is supported, indicating that overall AIDS incidence in Spain has already reached a maximum and has a tendency to remain stable or to decline in future. Long term expected values have become stable in most Communities; a slight increase is expected only in Extremadura. However, this Community has a relatively sparse population, and its contribution on the overall Spanish incidence is small. Long term expected values are estimated to be around 152.99 new cases per million inhabitants per year. This value is slightly smaller than the maximum incidence, observed in 1994 (179.4 cases). PMID- 11027957 TI - A survey of community gardens in upstate New York: implications for health promotion and community development. AB - Twenty community garden programs in upstate New York (representing 63 gardens) were surveyed to identify characteristics that may be useful to facilitate neighborhood development and health promotion. The most commonly expressed reasons for participating in gardens were access to fresh foods, to enjoy nature, and health benefits. Gardens in low-income neighborhoods (46%) were four times as likely as non low-income gardens to lead to other issues in the neighborhood being addressed; reportedly due to organizing facilitated through the community gardens. Additional research on community gardening can improve our understanding of the interaction of social and physical environments and community health, and effective strategies for empowerment, development, and health promotion. PMID- 11027959 TI - Accessing health care in a rural area: an evaluation of a voluntary medical transport scheme in the English Midlands. AB - In recent years notions of self-help and voluntarism have emerged as key elements in the delivery of services in rural England. This paper explores these themes by reference to 'Rural Wheels', a voluntary medical transport scheme in rural Northamptonshire, introduced to overcome the closure of branch surgeries and to provide access to a new medical centre. By focusing upon the organisation and operations of the scheme, the paper highlights the important role it plays in the welfare of rural residents, particularly elderly women. Yet, because effectively it is run by a small core group, the paper raises questions not just about the viability of this scheme but also about the increasing commitment of central government to the voluntary sector as a means of delivering health care to rural people. PMID- 11027958 TI - Regional equity in childhood health--register-based follow-up of the Finnish 1987 birth cohort. AB - Our objective was to investigate regional health differences among Finnish children using a population-based longitudinal register data. All live births born in 1987 were included in the study (N=59,546) and followed-up until the age of seven years. Statistically significant regional variation was found for all health indicators but diabetes. Background variables, such as maternal age and social class, explained only the difference in mortality. Various indicators gave different geographical patterns. Regional equity in childhood health has not been achieved in Finland. Existing health registers were feasible in studying regional variation in health, but a set of comprehensive morbidity indicators - preferably derived from different data sources - should be developed to monitor equity in health. PMID- 11027960 TI - Mortality differentials in the Czech Republic during the post-1989 socio political transformation. AB - The Czech Republic, together with Slovakia and Poland, forms a region within Central-Eastern Europe in which the values of life expectancy at birth have been increasing during the period of transformation. However, the tempo of mortality reduction has differed spatially within the territory of the Czech Republic, as have other outcomes of the transformation process. This paper discussed possible socio-economic explanations of regional differences in the tempos of mortality change between 1990/91 and 1995/96. Standardized mortality rates for males aged 0 64 years specified for the three most frequent causes of death were examined by means of the regression and correlation analysis. PMID- 11027961 TI - London's burning: recovering other geographies of health. AB - In the opening to this paper we concern ourselves with the concept of health as it is currently understood in the now reformulated 'geography of health'. We focus on countering the assumption that the desire for health is the main drive behind individual patterns of consumption. To this end, we turn to a more culturally informed analysis which identifies the importance of understanding the complex relations between the body, identity, consumption and risk. To illustrate our ideas and arguments, we draw on current anti-smoking debates, particularly those relating to passive smoking, and what we see as evidence of alternatives to such health-related or 'healthist' rhetoric: the rise of cigar smoking and cigar bars in London, England. PMID- 11027962 TI - Cancer chemotherapy and somatic cell mutation. AB - The occurrence of a second neoplasm is one of the major obstacles in cancer chemotherapy. The elucidation of the genotoxic effects induced by anti-cancer drugs is considered to be helpful in identifying the degree of cancer risk. Numerous investigations on cancer patients after chemotherapy have demonstrated: (i) an increase in the in vivo somatic cell mutant frequency (Mf) at three genetic loci, including hypoxanthine-guanine phosphoribosyl-transferase (hprt), glycophorin A (GPA), and the T-cell receptor (TCR), and (ii) alterations in the mutational spectra of hprt mutants. However, the time required for and the degree of such changes are quite variable among patients even if they have received the same chemotherapy, suggesting the existence of underlying genetic factor(s). Accordingly, some cancer patients prior to chemotherapy as well as patients with cancer-prone syndrome have been found to show an elevated Mf. Based on the information obtained from somatic cell mutation assays, an individualized chemotherapy should be considered in order to minimize the risk of a second neoplasm. PMID- 11027963 TI - A staining procedure for micronucleus test using new methylene blue and acridine orange: specimens that are supravitally stained with possible long-term storage. AB - The micronucleus test has been widely used as an in vivo cytogenetic test. It employs two different kinds of supravital staining methods which use either new methylene blue (N) and Giemsa (G) or acridine orange (AO). We have developed a new staining procedure for the preparation of specimens supravitally stained with possible long-term storage, using both N and AO. This N/AO-staining method involves three steps; (1) combination of the target tissue or target cells with an equivalent volume of 0.5% solution of new methylene blue (N-staining step), (2) immediate smear of the mixture, followed by treatment with methanol for 10 min for fixation and removal of N and drying (referred to as fixed-decolorized specimens), and (3) staining with 0.007% solution of AO for 3 min, followed by washing with Sorensen's buffer (pH 6.8) and covering of specimens before observation (AO-staining step). To examine whether the N/AO-staining method is useful for the micronucleus test, comparisons were made between N-, N/AO-, and AO stained specimens prepared supravitally from peripheral blood of rats with and without treatment of cyclophosphamide. The results indicate that N/AO-stained specimens can be supravitally observed after long-term storage with the same coloration and comparable frequencies of micronucleated reticulocytes with a positive response as AO-stained specimens, if the staining process is temporarily stopped before AO-staining (as fixed-decolorized specimens), or if the AO staining step is repeated. The results also showed that separated reticulocyte types are supravitally stained in a similar fashion to N-stained specimens but not to AO-stained specimens, indicative of the preservation of the supravital feature of N-staining. Taken together these results suggest that the N/AO staining procedure could offer an additional useful staining tool for the micronucleus test. PMID- 11027964 TI - Increased lymphocyte sister chromatid exchange frequency in workers with exposure to low level of ethylene dichloride. AB - The genotoxicity of low-level exposure to ethylene dichloride (EDC) and vinyl chloride monomer (VCM) in humans is not clear. We used lymphocyte sister chromatid exchange (SCE) frequency as a parameter to investigate the genotoxicity of low level EDC and VCM in VCM-manufacturing workers. The SCE frequency was determined for 51 male workers with exposure to VCM and/or EDC and for 20 male workers devoid of such exposure. Epidemiological data were obtained by questionnaire, and included history of smoking, drinking, and any medication taken, as well as a detailed occupational history. Personal- and area-sampling and analysis were conducted in order to calculate the time-weighted average (TWA) contaminant-exposure level corresponding to different job categories. Moderate EDC exposure around 1 ppm corresponded to a significantly greater SCE frequency than was the case for the low EDC exposure group (p<0.01). However, VCM exposure of similar level was not associated with increased SCE. We conclude that EDC may cause genotoxicity at a relatively low level of exposure. PMID- 11027965 TI - Activation of genotoxins to DNA-damaging species in exfoliated breast milk cells. AB - Exfoliated cells, isolated from breast milk samples donated by UK-resident women (n=15), were incubated, either immediately or after culture for 7 days, with one of a series of genotoxins, either in the presence or absence of the DNA-repair inhibitors, hydroxyurea (HU), and cytosine arabinoside (ara-C). The numbers of DNA single-strand breaks induced were then assessed as comet tail length (CTL) (microm) using the alkaline single cell-gel electrophoresis ('Comet') assay; cell viability was measured by trypan blue exclusion. The heterocyclic aromatic amines (HAAs) (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (0.4 mM), 3-amino 1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) (1.67 mM), 3-amino-1-methyl-5H pyrido[4,3-b]indole (Trp-P-2) (1.77 mM)), a polycyclic aromatic hydrocarbon (benzo[a]pyrene (B[a]P) (0.36 mM)), a nitro-polycyclic aromatic hydrocarbon (1 nitropyrene (1-NP) (1.84 mM)) and aromatic amines (o-toluidine (0.85 mM), p chloroaniline (0. 71 mM)) each induced statistically significant (P<0.0001, Mann Whitney test) increases in median CTLs in breast milk cells from all the donors examined when incubated (30 min, 37 degrees C) in the presence of HU/ara-C. In some cases, these compounds were also active in the absence of the repair inhibitors. There were marked variations in comet formation between donors and between genotoxins. Cell culture appeared to increase the epithelial cell proportion and cultured cells retained their ability to activate genotoxins. The results suggest that breast milk is a valuable source of human mammary cells for the study of the metabolic activation of possible carcinogens. PMID- 11027966 TI - Analysis of deletion mutations of the rpsL gene in the yeast Saccharomyces cerevisiae detected after long-term flight on the Russian space station Mir. AB - Using the yeast Saccharomyces cerevisiae on board the Russian space station Mir, we studied the effects of long-term space flight on mutation of the bacterial ribosomal protein L gene (rpsL) cloned in a yeast-Escherichia coli shuttle vector. The mutation frequencies of the cloned rpsL gene on the Mir and the ground (control) yeast samples were estimated by transformation of E. coli with the plasmid DNAs recovered from yeast and by assessment of the conversion of the rpsL wild-type phenotype (Sm(S)) to its mutant phenotype (Sm(R)). After a 40-day space flight, some part of space samples gave mutation frequencies two to three times higher than those of the ground samples. Nucleotide sequence analysis showed no apparent difference in point mutation rates between the space and the ground mutant samples. However, the greater part of the Mir mutant samples were found to have a total or large deletion in the rpsL sequence, suggesting that space radiation containing high-linear energy transfer (LET) might have caused deletion-type mutations. PMID- 11027967 TI - Postlabelling analysis of DNA adducts formed in human hepatoma cells treated with 3-nitrobenzanthrone. AB - 3-Nitrobenzanthrone (NBA) is one of the most mutagenic nitroaromatic compounds that has been found recently in diesel exhaust and airborne particles. A [32P] postlabelling analysis was carried out to examine the adducts in DNA from human hepatoma HepG2 cells treated with NBA. Two major and two minor adduct spots were obtained in the analysis. The structure of the compound obtained from one of the minor adduct spots was identified to be N-acetyl-3-amino-2-(2'-deoxyguanosin-3', 5'-bisphosphate-8-yl)-benzanthrone, based on identical mobility of the compound with that of synthetic standards in thin-layer chromatography and high performance liquid chromatography. This substance is the identical adduct found in our previous in vitro study. The yet-unidentified major adduct spots may be guanosin- and adenosin-benzanthrone adducts without the N-acetyl group. PMID- 11027968 TI - Chlorination of harman and norharman with sodium hypochlorite and co-mutagenicity of the chlorinated products. AB - Harman and norharman are widely distributed in the environment and consequently contaminate in domestic waste-water. It has been reported that they have co mutagenic activity in the presence of non- mutagenic aromatic amines such as aniline and o-toluidine with S9 mix. When these beta-carbolines were treated with sodium hypochiorite under mild conditions, chlorinated derivatives were produced. Among them, 6-chloroharman and 6-chloronorharman showed much more potent co mutagenic activities than harman and norharman in the presence of o-toluidine toward Salmonella typhimurium TA98 with S9 mix. These results suggest that the chlorination of harman and norharman occurs during disinfection at the sewage plant to produce potent co-mutagens that contaminate river water. PMID- 11027969 TI - Comparison of the mutations induced by p-benzoquinone, a benzene metabolite, in human and mouse cells. AB - Benzene is one of the chemicals widely contaminating the environment. Benzene is suggested to be a human leukemogen. When benzene is absorbed in the human body, it is metabolized firstly in the liver and subsequently in the bone marrow where it provokes initiation of leukemia. In the present study, we analyzed mutations induced by p-benzoquinone (p-BQ), a benzene metabolite, in human cells using a shuttle vector plasmid pMY189, and compared frequencies, types and spectra of the mutations with those of the mutations previously revealed in mouse cells using a similar plasmid pNY200. We found that p-BQ induces mutations in human and mouse cells at similar frequencies but with different types of mutagenesis. The proportion of tandem base mutations was significantly lower in human cells than in mouse cells. Most base substitutions were induced in G:C base pairs in both human and mouse cells. However, the proportion of G:C-->C :G transversion is significantly higher in human cells. These findings indicate that the p-BQ induced DNA damage in human and mouse cells is processed in a different manner, and that extrapolation of mice findings on experimental benzene carcinogenesis to human cancer risk assessment should be conducted carefully. PMID- 11027971 TI - Investigations on the mutagenicity of 1,4-dichlorobenzene and its main metabolite 2,5-dichlorophenol in vivo and in vitro. AB - The genotoxic potential of 1,4-dichlorobenzene (1,4-DCB) has been extensively evaluated in vitro and in vivo. The majority of the studies demonstrated the absence of a genotoxic potential for 1, 4-DCB. At variance are a bone marrow micronucleus test (MNT) after intraperitoneal (i.p.) treatment of NMRI mice [Mohtashamipur et al., Mutagenesis 2 (1987) 111-113] and a gene mutation assay on mouse lymphoma cells [McGregor et al., Environ. Mol. Mutagen. 12 (1988) 85-145]. Therefore, we investigated 1,4-DCB and its main metabolite 2,5-dichlorophenol (2,5-DCP) for both endpoints. In an MNT, male and female NMRI mice were treated orally with single doses of 2500mg/kg 1,4-DCB and 1500mg/kg 2,5-DCP, respectively. Smears were prepared 24, 48 and 72h thereafter. No induction of micronuclei was detected for both compounds. Also under the conditions of Mohtashamipur et al. (1987), intraperitoneal treatments of male and female mice with 2 x 177.5 and 2 x 355mg/kg 1,4-DCB failed to induce micronuclei. In addition, CHO/HPRT-gene mutation tests with 1,4-DCB and 2,5-DCP yielded negative results for both compounds with and without metabolic activation system. Therefore, 1,4-DCB and 2,5-DCP are considered to be non-mutagenic in these test systems. PMID- 11027970 TI - Plant activation of aromatic amines mediated by cytochromes P450 and flavin containing monooxygenases. AB - To know the mechanisms involved in the activation of promutagenic aromatic amines mediated by plants, we used Persea americana S117 system (S117) for the activation of 2-aminofluorene (2-AF) and m-phenylenediamine (m-PDA) in Ames assays. In these assays, the effect of the diphenylene iodonium (DPI), an inhibitor of flavin-containing monooxygenases (FMOs), of the 1-aminobenzotriazole (1-ABT), an inhibitor of cytochromes P450 (cyt-P450s) and of the methimazole, a high-affinity substrate for FMOs, was studied. The efficacy of both inhibitors and of the methimazole was verified to find that they did partially inhibit the mutagenesis of both aromatic amines, activated with rat liver S9. Similarly, both inhibitors and methimazole did produce a significant decrease in 2-AF and m-PDA mutagenesis, when the activation system was S117, indicating that, similar to what occurs in mammalian systems, plant FMOs and cyt-P450s can metabolize aromatic amines to mutagenic product(s). However, the affinity of both FMOs and cyt-P450s of plant for 2-AF and m-PDA was different. Data obtained indicate that the activities of plant FMOs must be the main enzymatic system of m-PDA activation while, in 2-AF activation, plant cyt-P450s have the most relevant activities. In addition, peroxidases of the S117 system must contribute to 2-AF activation and some isoforms of FMOs and/or cyt-P450s of the S117 system, uninhibited by the inhibitors used, must be the responsible for a partial activation of m-PDA. PMID- 11027972 TI - Conventional armament wastes induce micronuclei in wild brown trout Salmo trutta. AB - We analysed micronuclei in brown trout Salmo trutta specimens sampled in the Trubia River, upstream and downstream of the emissions from a Spanish military factory to assess genotoxicity risks derived from military wastes. A significant exponential increase in micronuclei counts was found in fish living downstream of the military wastes with respect to fishes inhabiting upstream areas of the same river. In comparison, we only found a linear increase in micronuclei counts in a control stream where an old military factory had been demolished 6 months before sampling. This difference suggests that active discharge of armament factory wastes can directly induce micronuclei and therefore represents a genotoxic risk for the ecosystem. PMID- 11027973 TI - Mutagenic properties of a nitrofuran, 7-methoxy-2-nitronaphtho[2, 1-b]furan (R7000), in lacI transgenic mice. AB - The in vivo mutagenic properties of a 5-nitrofuran, the 7-methoxy-2 nitronaphtho[2,1-b]furan (R7000), already well known in bacteria, was evaluated in lacI transgenic mice (Big Blue). The mutation frequency was determined in various organs of i.p. - treated mice and the nature of induced mutations was determined for the target organs in which mutation induction was significant. It was found that R7000 is mutagenic in mice, although, on the basis of the number of induced mutants per unit mass in comparison with other known mutagenic chemicals, R7000 appears to be considerably less mutagenic in mice than in bacteria. The most affected organs, small intestine, caecum and colon organs belong to the digestive apparatus. The distribution of R7000-induced mutations in the lacI gene recovered from small intestine of transgenic mice was very similar to that which had been found in E. coli. The difference between mouse and E. coli in the R7000 induced mutational spectra are mainly in the proportion of single base frameshifts versus base substitutions. Since R7000 induced mutations seemed to arise in the population of stem cells and that the stem cells are important for carcinogenesis, our results are compatible with a possible carcinogenic effect of R7000 and other nitrofurans. PMID- 11027974 TI - Dietary restriction during murine development provides protection against MNU induced mutations. AB - The developmental stage is the most rapid period for the accumulation of somatic mutations. Epidemiological studies have also suggested a significant role of early life for cancer susceptibility, showing a protective effect of modest dietary restriction early in life. To determine if mutation rate, diet, and cancer risk are related, we have investigated the effect of dietary restriction on somatic mutations early in life. The diet of mouse dams was restricted during pregnancy and lactation by 10% from ad libitum control. F(1) pups (SWRxMutaMouse) were weaned at 3 weeks of age. Pups from dams that were on a restricted diet were kept under dietary restriction (40% until 5 weeks of age and then 20% until sacrifice). Only females from litters of seven or eight were used in this study. A portion of pups from both groups were treated with N-methyl-N-nitrosourea (MNU, 50mg/kg, i.p.) at 5 weeks of age and all mice were sacrificed at 10 weeks of age. The frequency of induced mutations was reduced by about 30% at the three loci studied, lacZ (P=0.028) and cII (P=0.042) and Dlb-1 (P=0.032) in the small intestine in the restricted group. A similar decrease in the lacZ mutant frequency was observed in the bone marrow, but the results did not reach statistical significance (P=0.074). Few differences in the lacZ mutant frequency were observed in the colon and the mammary epithelium, but variability of the mutant frequencies was such that an effect of similar magnitude could not be excluded statistically. Analysis of 47 cII mutants revealed that the majority of MNU-induced mutations were G:C to A:T transition at non-CpG sites, with no difference in the mutation spectrum between the two dietary groups. PMID- 11027975 TI - Increased lymphocyte DNA strand breaks in rubber workers. AB - OBJECTIVE: To study the effect of occupational exposure to rubber processing, smoking, and alcohol drinking on lymphocyte DNA damage. SUBJECTS AND METHODS: Of 371 employees (197 men and 174 women) from a rubber factory in Guangzhou, 281 were rubber processing workers from five production sections and 90 were managerial workers. Information on occupational exposure, smoking, and drinking was collected by interviews. Blood samples were taken in the morning by venipuncture. DNA damages were measured by the Comet assay. Possible DNA-protein crosslinks were broken down by proteinase K. Tail moment, measured by Komet 4.0 image analysis software, was the measure of DNA damage. RESULTS: The rubber processing workers had larger tail moment than the managerial workers (Geometric mean, 95%CI) [1. 77microm (1.64-1.90) versus 1.52microm (1.36-1.71), P=0.04]. Both smoking [1.93microm (1.74-2.13) versus 1.59microm (1.47-1.71), P=0. 003] and alcohol drinking [2.21microm (1.87-2.62) versus 1.63microm (1.53-1.74), P<0.001] increased tail moment. Tail moment differed significantly among job categories (F=3.21, P=0.008), the largest was observed in mixers. In the non-smoking and non drinking workers, rubber processing workers had larger tail moment than managerial workers after adjusting for age (P=0.033). General linear model analysis showed that after adjusting for each other, occupational exposure (P=0.027), smoking (P=0.012), and alcohol drinking (P=0. 013) was associated with larger tail moment, whereas age and gender had no effect. CONCLUSIONS: Occupational exposure to rubber processing, smoking, and alcohol drinking can cause DNA damage. PMID- 11027976 TI - Comparison of micronuclei frequencies in mono-, bi- and poly-nucleated lymphocytes from subjects of a residential suburb and subjects living near a metallurgical plant. AB - Spontaneous baseline frequencies of micronuclei in mono-, bi- and poly-nucleated lymphocytes were analyzed, using the cytokinesis-block technique, in 103 subjects living in a residential suburb (Genova-Nervi), and in 203 subjects living in an urban industrialized area near a metallurgical plant and a coke factory (Genova Cornigliano). Statistical analysis showed that the average frequency of micronucleated binucleated lymphocytes (MnBNL) was significantly higher (1.42 fold) in donors of Nervi than in donors of Cornigliano living in a contaminated environment. In contrast, the average frequency of micronucleated polynucleated lymphocytes (MnPNL) was significantly higher (1.66-fold) in donors of Cornigliano than in donors of Nervi. The existence in the whole population examined of a positive correlation between frequency of MnBNL and frequency of MnPNL and the absence of a positive correlation between frequency of bi- and poly-nucleated lymphocytes and frequency of MnPNL suggest that the formation of MnPNL is a consequence of genetic damage and not of mitotic errors arising during the division of bi- and poly-nucleated cells. In agreement with previous findings the frequency of MnBNL increased with age and was significantly higher in females than in males; unexpectedly it was higher in non-smokers/non-drinkers than in smokers/drinkers. PMID- 11027977 TI - Genotoxicity of the coating lacquer on food cans, bisphenol A diglycidyl ether (BADGE), its hydrolysis products and a chlorohydrin of BADGE. AB - The epoxy resin bisphenol A diglycidyl ether (BADGE), its hydrolysis products and a chlorohydrin of BADGE (BADGE.2HCl), were examined for their genotoxicity in the micronucleus test (MNT) with human peripheral blood lymphocytes in vitro, in presence and in absence of an exogenous metabolizing system S9 rat liver. The treatment was done using different compound concentrations up to cytotoxic doses. The concentrations tested ranged between 12.5 to 62.5microg/ml of BADGE, 12.5 to 62.5microg/ml of first BADGE hydrolysis product (BADGE.H(2)O), 25.0 to 100.0microg/ml of second BADGE hydrolysis product (BADGE.2H(2)O) and 6.25 to 50.0microg/ml of BADGE.2HCl. These compounds are able to induce both cytotoxic and genotoxic effects, as revealed by the increases observed in cytokinesis block proliferation index (CBPI) and in micronuclei (MN) frequencies, respectively. PMID- 11027978 TI - The apoptotic pathway: a target for therapy in chronic lymphocytic leukemia. AB - Cell division and apoptosis (programmed cell death) are the two major physiological processes which control the size of cell populations. Chronic lymphocytic leukaemia arises as a result of the clonal expansion of, usually B-, lymphocytes in which a dysregulation of apoptosis leads to prolonged cell survival. The same process becomes exaggerated with increasing drug resistance, the usual cause of treatment failure in this condition. The identification of points in the apoptotic pathway at which dysregulation occurs is beginning to open up new therapeutic opportunities where the conventional cytotoxic chemotherapy approach is found to fail. Although these strategies are still in their infancy they may increase understanding of the pathogenesis of the disorder and overcome the problem of drug resistance. PMID- 11027979 TI - DNA repair mechanisms and acute myeloblastic leukemia. AB - DNA repair mechanisms play a vital role in maintaining genomic integrity. With the wealth of knowledge gained recently on these processes it is becoming clear that defects in repair proteins and proteins associated with the regulation of repair are connected to many different human diseases including cancer. This paper has aimed to review the four major DNA repair processes and in particular concentrate on their association with acute myeloblastic leukemia (AML). PMID- 11027980 TI - Positive and negative selection to reduce tumour contamination in peripheral blood stem cell harvests. AB - Peripheral blood progenitor cells used during high dose treatments for malignancy may be contaminated with tumour cells that could later contribute to recurrence. CD34+ selected harvests still contain tumour cells and an additional negative selection may be capable of reducing this contamination. We have assessed a two stage technique in which a CD34+ selection is followed by a tumour specific depletion stage using a B cell or breast cancer specific antibody panel. Initial small-scale selections on 11 patients with NHL and breast cancer showed that cell loss was greatest following the CD34+ selection with a median yield of 38.8 per cent (range 17. 2-56.4 per cent). The addition of the depletion stage resulted in a minimal loss of CD34+ cells with a yield for this step of 94.2 per cent (range 77.5-99.3 per cent). Clinical scale selections were performed on seven patients with CLL and a median of 2.8x10(6)/kg CD34+ cells (range 1.5-6.1x10(6)/kg) were collected. Cell recovery was 53.3 per cent following CD34+ selection and 76.9 per cent following the tumour specific depletion stage, resulting in a final product containing a median of 1.0x10(6)/kg CD34+ cells (range 0. 55-2.0x10(6)/kg). All unmanipulated harvests were heavily contaminated with tumour cells (median contamination 10.2 per cent, range 2.0-83.1 per cent) as measured by flow cytometry and a median 4.7 log (range 3-5 log) tumour cell purge was produced following two-stage selection. Six of the patients have received cells manipulated in this way with median engraftment times of neutrophils>0.5x10(9)/l=16 days (range 13-20 days) and platelets>20x10(9)/l=16.5 days (range 11-42 days). At a median follow-up of 25 months, these transplanted patients remain well and in molecular complete remission. PMID- 11027981 TI - Recent publications in hematological oncology. PMID- 11027983 TI - Enzyme Mechanisms for Polycyclic Triterpene Formation. AB - The mechanisms by which triterpene cyclases transform olefins into complex and biologically important polycyclic products have fueled nearly half a century of intense research. Recent chemical and biological studies, together with previous findings, provide intriguing new insights into the enzymatic mechanism of triterpene formation and form a surprisingly detailed picture of these elegant catalysts. It can be concluded that the role of the oxidosqualene cyclases involves protection of the intermediate carbocation against addition of water or deprotonation by base, thereby allowing the shift of the hydride and methyl groups along a thermodynamically and kinetically favorable cascade. Key questions in the areas of structural biology, site-directed mutagenesis, and directed evolution are apparent, now that the first structure of a triterpene cyclase is known. PMID- 11027985 TI - A New Role for Polyketides. PMID- 11027984 TI - The Interleukin-4-Receptor: From Recognition Mechanism to Pharmacological Target Structure. AB - Organic synthesis of hormone derivatives is an established route to yield pharmacologically active agents. Until recently this has only been feasible for small organic compounds, but nowadays it is also possible to produce antagonists for larger protein hormones. In particular, the interleukin-4-receptor was a well suited target for this approach since it plays a pivotal role in the release and progression of allergic diseases. Accordingly, a strong interest and a high medical need is associated with the development of inhibitors. The structural elucidation of the ligand/receptor complex and an improved understanding of the mechanisms concerning receptor binding and activation allow for the rational design of variants that inhibit interleukin-4. Since it is possible to specifically inhibit the interleukin-4-receptor system in this way, a completely new approach to the development of new drugs against allergy and asthma has been established. PMID- 11027986 TI - D-Serine as a Modulator in the Nervous System. PMID- 11027987 TI - A Demonstration of the Primary Stereoelectronic Effect in the Baeyer-Villiger Oxidation of alpha-Fluorocyclohexanones The Natural Sciences and Engineering Research Council of Canada (NSERC) is gratefully acknowledged for support of this research in terms of research grants to C.C. and PGS scholarships to A.C.C. We would especially like to thank Professor Scott E. Denmark, Dr. Robert Syvret, and Dr. Claudio Sturino for helpful discussions. PMID- 11027988 TI - Dendritic Growth of a Supramolecular Complex We acknowledge financial support from the Xunta de Galicia (Projects XUGA 26201b96 and XUGA PGIDT99PXI26201B) and CYTED (Project VIII.3). E.A.P. and P.R.C. thank CONACYT (Mexico) and Xunta de Galicia (Spain) for research scholarships. beta-Cyclodextrin was kindly supplied by Roquette, France. PMID- 11027989 TI - Lithium Binaphtholates: Chiral Chains and Clusters This work was supported by the EPSRC (studentship to J.L.M.D.). PMID- 11027990 TI - A Model for the Nonenzymatic BCD Cyclization of Squalene This research was supported by FWO-Vlaanderen. We thank Dr. Davide Proserpio (Universita di Milano) and Dr. Annalisa Guerri (Universita di Firenze) for the allowance to use the CCD diffractometers. PMID- 11027991 TI - A Simple Model for the Origin of Enantioselection and the Anti "Lock-and-Key" Motif in Asymmetric Hydrogenation of Enamides as Catalyzed by Chiral Diphosphine Complexes of Rh(I) This work was supported by the National Science Foundation (CHE-9618497) and by the National Center for Supercomputer Applications under MCA99S017N and used the CRAY ORIGIN 2000 computer system at the NCSA. PMID- 11027992 TI - Stepwise Ethene and/or Methyl Acrylate/CO Insertions into the Pd-C Bond of Cationic Palladium(II) Complexes Stabilized by a (P,O) Chelate This work was supported by the Centre National de la Recherche Scientifique, the Ministere de l'Education Nationale, de l'Enseignement et de la Recherche (Grant to C.F. and Action Incitative), Elf Atochem, and Elf Aquitaine. Special thanks are due to Ms. N. Kyritsakis, Dr. A. DeCian, and Prof. J. Fischer for the X-ray structure determination of 3. We also thank Dr. R. Graff (Service Commun RMN, Universite Louis Pasteur) and Mr. R. Hubert (Laboratoire de Spectrometrie de Masse, Universite Louis Pasteur). PMID- 11027993 TI - An Efficient, Modular Synthetic Route to Oligomers Based on Zirconocene Coupling: Properties for Phenylene-Thiophene-1-Oxide and Phenylene-Thiophene-1,1-Dioxide Chains This research was supported by the National Science Foundation. M.C.S. thanks the Korea Science and Engineering Foundation (KOSEF). PMID- 11027994 TI - Vitronectin Receptor Antagonists: Purine-Based Peptidomimetics. PMID- 11027995 TI - Highly Efficient and Versatile Acylation of Alcohols with Bi(OTf)(3) as Catalyst. PMID- 11027996 TI - Remarkable In/out Inversions at Bridgehead Phosphorus Atoms We thank Dr. J. M. Oliva (Institute of Materials Sciences of Barcelona, CSIC) for the calculations on 12 and 13, the EPSRC for a quota studentship to D.R., and the EPSRC National Mass Spectrometry Service for high-resolution electrospray spectra. PMID- 11027997 TI - Gp120 Binds Cooperatively to Several Biologically Relevant Glycosphingolipids: Quantitative Measurements at Equilibrium by Total Internal Reflection Fluorescence Microscopy This research was supported by the NIH (AI40359-02), the NSF (CHE-9726132 and CHE-9623583), Eli Lilly (JGH), and the Alfred P. Sloan Foundation (J.G.H.). K.D.M. gratefully acknowledges receipt of the University of Arizona Dean's Fellowship and the Department of Chemistry Carl S. Marvel Fellowship. We thank Ying-Mei Gu for performing the streptavidin adsorption measurements. PMID- 11027998 TI - A Mixed-Valence Mixed-Spin Prussian-Blue-Like Heptanuclear Complex We are indebted to Prof. P. Day of The Royal Institute of Great Britain for interesting discussions that were at the origin of the work presented in this paper. PMID- 11027999 TI - Integer-Spin Multifrequency EPR Spectroscopy of a Ferromagnetically Coupled, Oxo Bridged Mn(IV)Mn(IV) Model Complex We thank Dr. S. Gambarelli for the Q-band EPR spectrum. PMID- 11028000 TI - Luminescent Mesoporous Molecular Materials Based on Neutral Tetrametallic Rectangles We gratefully acknowledge the U.S. National Science Foundation and the National Oceanic and Atmospheric Administration for financial support. Mass spectrometry was provided by the Washington University Mass Spectrometry Resource, an NIH Research Resource (Grant No. P41RR0954), and by the UIUC School of Chemical Science. PMID- 11028001 TI - Enantioselective Synthesis of 4-Unsubstituted 3-Alkoxy- and 3-Aminoazetidin-2 ones from Formaldehyde N,N-Dialkylhydrazones We thank the Direccion General de Investigacion Cientifica y Tecnica for financial support (Grant no. PB 97/0747). We also thank the Ministerio de Educacion y Ciencia for a doctoral fellowship to A.F. PMID- 11028002 TI - A Highly Conductive Macrocycle-Linked Metallophthalocyanine Polymer Funding from the Office of Naval Research is gratefully acknowledged. This work was supported in part by the MRSEC Program of the National Science Foundation under award number DMR 98-08941. PMID- 11028003 TI - Symmetrical Scission of the Coordinated Tetraborane in PMID- 11028004 TI - Highly Efficient Ring-Opening Metathesis Polymerization (ROMP) Using New Ruthenium Catalysts Containing N-Heterocyclic Carbene Ligands C.B. is grateful to the National Science Foundation for a pre-doctoral fellowship. The authors thank Dr. Matthias Scholl for providing catalysts 4 a and 4 c. PMID- 11028005 TI - The Phenylsulfonyl Group as an endo Stereochemical Controller in Intramolecular Pauson-Khand Reactions of 3-Oxygenated 1,6-Enynes Financial support of this work by the Ministerio de Educacion y Cultura (DGES, project PB96-0021) and Comunidad de Madrid (project 07B/28/1999) is gratefully acknowledged. PMID- 11028006 TI - Cyclization of Terminal Diynes Catalyzed by Thiolate-Bridged Diruthenium Complexes: A Simple Synthetic Route to endo-Macrocyclic (Z)-1-En-3-ynes This work was supported by a Grant-in-Aid for Specially Promoted Research (09102004) from the Ministry of Education, Science, Sports, and Culture, Japan. PMID- 11028007 TI - A Semiconducting Lamella Polymer PMID- 11028008 TI - Controlled Assembly of Polyoxometalate Chains from Lacunary Building Blocks and Lanthanide-Cation Linkers Supported in part by the National Science Foundation (CHE9727417) and Georgetown University. PMID- 11028009 TI - Halide-Ion Encapsulation by a Flexible Dicopper(II) Bis-Tren Cryptate. PMID- 11028010 TI - Unprecedented Expansion of the Pore Size and Volume of Periodic Mesoporous Silica This was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC). Thanks go to Y. Yang for technical help. PMID- 11028011 TI - Template Synthesis of the First 1,4,7-Triphosphacyclononane Derivatives This work was funded by the UK Engineering and Physical Sciences Research Council. PMID- 11028012 TI - Through-Space Exciton Coupling and Multimodal Na(+)/K(+) Sensing Properties of Calix PMID- 11028013 TI - In Situ (1)H MAS NMR Spectroscopic Observation of Proton Species on a Mo-Modified HZSM-5 Zeolite Catalyst for the Dehydroaromatization of Methane We are grateful for the support of the National Natural Science Foundation of China and the Ministry of Science and Technology of China. PMID- 11028014 TI - First Crystal Structure of a Medicinally Relevant Gold Protein Complex: Unexpected Binding of PMID- 11028015 TI - DeNOx of Exhaust Gas from Lean-Burn Engines through Reversible Adsorption of N(2)O(3) in Alkali Metal Cation Exchanged Faujasite-Type Zeolites This work was sponsored by the European Community (Brite Euram III projects BE-95-2127 "SNR Technique" and BE-97-4493 "SORPTEC") and the Belgium government (IUAP-PAI program). PMID- 11028016 TI - Pronounced Steric Hindrance for Nitrogen Inversion in 1,3,4-Oxadiazolidines Asymmetric Nitrogen, Part 83. This work was supported by the Russian Foundation for Basic Research (RFBR) (grant No. 97-03-33021) and DFG-RFBR (grant No. 98-03 04119). O.T. thanks the Fonds der Chemischen Industrie for a doctorate scholarship. Part 82: D. A. Lenev, K. A. Lyssenko, R. G. Kostyanovsky, Izv. Akad. Nauk. Ser. Khim. 2000, N7 PMID- 11028017 TI - Stable and Selective Hybridization of Oligonucleotides with Unnatural Hydrophobic Bases This work was supported by the National Institutes of Health (GM 60005 to F.E.R.) and the Skaggs Institute for Chemical Biology (F.E.R. and P.G.S.). A.K.O. thanks the National Institutes of Health for a postdoctoral fellowship (F32 GM19833-01). PMID- 11028018 TI - Synthesis of a Complexed 2,2-Bisphosphirane This work was supported in part by the Council of Chemical Sciences with financial aid from the Netherlands Organization for Scientific Research (CW-NWO). PMID- 11028019 TI - Micellization versus Cyclodextrin-Surfactant Complexation Financial support from the Direccion General de Ensenanza Superior of Spain (project PB96-0954) and Xunta de Galicia (project PGIDT99 PXI30104B) is gratefully acknowledged. J. P.-J. wishes to thank the Ministerio de Educacion y Cultura for a F.P.U. fellowship. PMID- 11028020 TI - Domino Michael Aldol and Domino Michael Mannich Reactions: Highly Stereoselective Synthesis of Functionalized Cyclohexanes This work has been supported by the Deutsche Forschungsgemeinschaft (Schn 441/1-2), the Fonds der Chemischen Industrie (doctoral fellowship to O. R.) and Degussa Huls AG. We thank Prof. L. Tietze for constant support. PMID- 11028021 TI - Deoxygenative Cycloaddition of Aldehydes with Alkynes Mediated by AlCl(3) and Zirconium: Formation of Cyclopentadiene Derivatives Financial support from the National Natural Science Foundation of China (29702001), the National Science Fund for Distinguished Scholars (29825105), and the Peking University President Fund are gratefully acknowledged. Yunhai Xiao carried out some experimental work. PMID- 11028022 TI - Synthesis and Structure of an Anionic Ga(26)R(8) Cluster with a Metalloid Core The Chemistry of Gallium, Part 17. This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. We thank E. Mollhausen (Universitat Karlsruhe) for the collection of the crystallographic data and Priv.-Doz. Dr. B. Pilawa (Universitat Karlsruhe) for measuring an EPR spectrum. Part 16: ref. 6. PMID- 11028024 TI - Odds and Trends: Recent Developments in the Chemistry of Odorants Note on trademarks: Words which we know or have reason to believe constitute registered trademarks (R) are designated as such. However, neither the presence nor absence of such designation should be regarded as affecting the legal status of any trademark. Note on perfume analysis: The quoted percentages of perfume raw materials in market products are rounded figures. They are often derived from area percentages from the GC (FID) analysis, and are thus subject to analytical error. AB - Fragrance chemistry is, together with the closely related area of flavor chemistry, one of the few domains, if not the only one, in which chemists can immediately experience structure-activity relationships. This review presents structure-odor correlations and olfactophore models for the main odor notes of perfumery: "fruity", "marine", "green", "floral", "spicy", "woody", "amber", and "musky". New trendsetters and so-called captive odorants of these notes are introduced, and recent activities and highlights in fragrance chemistry are summarized. The design of odorants, their chemical synthesis, and their use in modern perfumery is discussed. Our selection is guided and illustrated by creative fragrances, and features new odorants which encompassed current trends in perfumery. New odorants for grapefruit and blackcurrant, for galbanum, and leafy top notes are presented. Compounds with fashionable marine, ozonic, and aquatic facets are treated, as well as new odorants for classical lily-of-the valley, rose, and jasmine accords. Compounds with sweet and spicy tonalities are also discussed, as are the most recent developments for woody notes such as sandalwood and vetiver. We conclude with musky and ambery odorants possessing uncommon or unusual structural features. Some odor trends and effects are illustrated by microencapsulated fragrance samples, and areas where there is need for the development of new synthetic materials and methodologies are pointed out. Thus, chemists are invited to explore fragrance chemistry and participate in the design and synthesis of new odorants. This review gives the latest state of the art of the subject. PMID- 11028025 TI - Olefin Metathesis and Beyond A list of abbreviations can be found at the end of this article. AB - The advent of well-defined catalysts for olefin metathesis which combine high activity, durability, and excellent tolerance towards polar functional groups has revolutionized the field. The past decade has seen the rapid embrace of these reagents as tools for advanced organic and polymer chemistry and the success of this development is witnessed by a plethora of elegant applications to the synthesis of natural and nonnatural products. This review article provides an overview of these developments and intends to familiarize the reader with some very recent advances which hold the promise to expand the scope of the reaction even further. Moreover, the positive impact of metathesis on the fundamental logic of retrosynthetic planning is demonstrated by means of typical examples. Finally, it will be shown that metathesis is by no means restricted to alkenes as substrates, and some comments on metathesis reactions following unconventional mechanistic pathways will also be presented. PMID- 11028026 TI - Kekule Escapes, Popper Notwithstanding I thank R. Hoffmann, J. M. McBride, and J. S. Siegel for helpful suggestions. PMID- 11028027 TI - New Strategies for the Development of an Asymmetric Version of the Baylis-Hillman Reaction This work was supported by the Fonds der Chemischen Industrie (Liebig scholarship and funds for P. L.). P. L. thanks Prof. Dr. A. de Meijere for his support. PMID- 11028028 TI - Nanoporous Structures by Design. PMID- 11028029 TI - Supramolecular Nanocircles Consisting of Streptavidin and DNA This work was supported by the Deutsche Forschungsgemeinschaft (SPP 1072), the Fonds der Chemischen Industrie, and the Tonjes-Vagt Stiftung. We thank Prof. D. Blohm and Prof. H. Fuchs for generous support. PMID- 11028030 TI - Dynamic Observations of the Hydrolysis of a DPPC Monolayer at the Air/Water Interface Catalyzed by Phospholipase A(2) This work was supported by the research contract between the German Max-Planck-Society and the Chinese Academy of Sciences as well as the National Natural Science Foundation of China (NNSF). J.L. thanks the president fund of the Chinese Academy of Science and the National Personal Department of China. DPPC=L-dipalmitolphosphatidylcholine. PMID- 11028031 TI - Uranium Hexakisamido Complexes This work was supported by the National Science Foundation (CAREER Award CHE-9501992), the Alfred P. Sloan Foundation, the National Science Board (1998 Alan T. Waterman award to C.C.C.), and the Packard Foundation. K.M. thanks the Deutsche Forschungsgemeinschaft for a postdoctoral fellowship. PMID- 11028032 TI - Base-Induced Disproportionation of Elemental Gold The authors are grateful to the Fonds der Chemischen Industrie for their financial support. A.-V. M. thanks the Studienstiftung der Hoechst AG, as well as the Ev. Studienwerk Villigst e.V. for their support. PMID- 11028034 TI - A Highly Active Catalyst System for Intermolecular Hydroacylation This work was supported by the Brain Korea 21 Project. PMID- 11028033 TI - Unusual C-H small middle dot small middle dot small middle dotSe=C Interactions in Aldols of Chiral N-Acyl Selones Detected by Gradient-Selected (1)H-(77)Se HMQC NMR Spectroscopy and X-ray Crystallography This work was supported by the Los Alamos National Laboratory Directed Research and Development Fund (grant XAB2). We thank Dr. Judith Gallucci (Department of Chemistry, Ohio State University) for obtaining the X-ray structures of 1 and 3, and Dr. Charles Campana (Bruker Analytical X-ray Systems in Madison, WI) for obtaining the X-ray structure of 2. PMID- 11028035 TI - Asymmetric Synthesis of a Chiral Secondary Grignard Reagent This study was supported by the Deutsche Forschungsgemeinschaft (SFB 260 and Graduiertenkolleg "Metallorganische Chemie") and the Fonds der Chemischen Industrie. PMID- 11028036 TI - Unusually Stable Vinyl Cations This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie (Ulm). The Frankfurt group thanks Prof. N. Auner for support. We are indebted to Prof. Mark Fink, Tulane University, New Orleans, for carefully reading our manuscript. We thank Thomas Nau, Computer Center Universitat Ulm, for adaptation of the Gaussian Program Suite. PMID- 11028037 TI - [] PMID- 11028038 TI - Silylated Cyclohexadienes: New Alternatives to Tributyltin Hydride in Free Radical Chemistry We are grateful to Prof. Dr. Dieter Seebach for generous financial support and to Prof. Dr. Erick M. Carreira for helpful discussions during the preparation of the manuscript. We also thank the Swiss Science National Foundation (2100-055280.98/1) for funding our work. PMID- 11028039 TI - [{Au[u-N] PMID- 11028040 TI - Umpolung of P-H Bonds This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. PMID- 11028041 TI - Stereoselective Synthesis and Palladium-Catalyzed Transformations of 2-Alkylidene 5-vinyltetrahydrofurans This work was supported by the Fonds der Chemischen Industrie (Liebig scholarship and funds for P.L.) and by the Deutsche Forschungsgemeinschaft. P.L. thanks Prof. Dr. A. de Meijere for his support. PMID- 11028042 TI - An Unusual Open-Framework Cobalt(II) Phosphate with a Channel Structure That Exhibits Structural and Magnetic Transitions The authors thank Professor S. K. Malik of the TIFR for help with the SQUID measurements. PMID- 11028043 TI - Formation of Extended Tapes of Cyclic Water Hexamers in an Organic Molecular Crystal Host We thank Prof. J. E. Jackson (MSU) for his generous support of this study. PMID- 11028044 TI - The First Solid-Phase Synthesis of a Peptide-Tethered Platinum(II) Complex This research was supported by the Council for Chemical Sciences of The Netherlands Organization for Scientific Research (CW-NWO) and by The Netherlands Foundation for Technical Sciences (STW). Support and sponsorship by COST Action D8/00097 (biocoordination chemistry) is kindly acknowledged. The authors thank Johnson & Matthey (Reading, UK) for their generous loan of K(2)PtCl(4). PMID- 11028045 TI - The First Structurally Characterized Aluminum Compounds with Terminal Acetylide Groups This work was supported by the Deutsche Forschungsgemeinschaft. N.C.M.-Z. thanks the Schweizerischer Nationalfonds for a fellowship. PMID- 11028046 TI - Ruthenium-Catalyzed Enyne Metathesis of Acetylenic Boronates: A Concise Route for the Construction of Cyclic 1,3-Dienylboronic Esters. PMID- 11028047 TI - A Light-Modulated Sequence-Specific DNA-Binding Peptide This work was supported by the Spanish M.E.C. (PB97-0524) and the Xunta de Galicia (XUGA 20906B96). A.M.C and E.V. thank the Xunta de Galicia and the University of Santiago for their predoctoral fellowships. We are grateful to Prof. M. Mosquera and Prof. J. Benavente for allowing us to use the spectrofluorimetry and radioactivity facilities, respectively, and to Dr. Chris Abell (Cambridge University, Iberdrola Visiting Professor at the University of Santiago) for critical reading of the manuscript. PMID- 11028048 TI - TEM Studies of Platinum Nanowires Fabricated in Mesoporous Silica MCM-41 A part of this work was supported by CREST, Japan Science and Technology Cooperation. PMID- 11028049 TI - Network Polysilanes: Synthesis, Electrical Conductivity, Charge-Transfer Interaction, and Photoconductivity. PMID- 11028050 TI - Synthesis, Structure, and Magnetism of a Novel Alkoxide Bridged Nonacopper(II) (Cu(9)O(12)) PMID- 11028051 TI - Highly Efficient Degradation of Thiophosphate Pesticides Catalyzed by Platinum and Palladium Aryl Oxime Metallacycles Financial support from the Russian Foundation for Basic Research (grant no. 98-03-33023a) and INTAS (project 97 0166) is gratefully acknowledged. PMID- 11028052 TI - [{] PMID- 11028053 TI - Asymmetric Alkylation of Nitroalkanes We thank the National Science Foundation and the National Institutes of Health (NIH), General Medical Sciences, for their generous support of our programs. Rhone-Poulenc graciously provided a postdoctoral fellowship for J.-P. S. Mass spectra were provided by the Mass Spectrometry Facility of the University of California, San Francisco, supported by the NIH Division of Research Resources. PMID- 11028054 TI - Enantiomeric Self-Recognition: Cation-Templated Formation of Homochiral Isoguanosine Pentamers This research was sponsored by the Separations and Analysis Program, Chemical Sciences Division, Office of Basic Energy Sciences, U.S. Department of Energy. J.D. thanks the Dreyfus Foundation for a Teacher Scholar Award. We thank Professors Giovanni Gottarelli and Steve Rokita for helpful discussions. PMID- 11028055 TI - Synthesis and Characterization of Iron Silasesquioxane Phosphane Complexes This work was supported by the Office of Basic Energy Sciences (U.S. DOE, contract no. W-7405-ENG-36 with the University of California) and by the LANL Laboratory Directed Research and Development Program. PMID- 11028056 TI - A Graphite-Like Complex with Large Cavities Constructed with the Complex Ligand PMID- 11028057 TI - A Parallel Library of all Seven A(2)+B(2)+C(2) T(h) Regioisomeric Hexakisadducts of Fullerene C(60): Inspiration from Werner's Octahedral Stereoisomerism This work was supported by a National Science Foundation Young Investigator Award (CHE 9457693), the Office of Naval Research (N00014-98-1-0035), and an Alfred P. Sloan Research Fellowship award. PMID- 11028058 TI - A Chip-Based Biosensor for the Functional Analysis of Single Ion Channels We thank E. Ermanntraut, L. Giovangrandi, T. Wohland, A. Brecht, M. Kohler, C. Bieri, D. Stamou, and R. Hovius for advice. This work was supported by the Swiss National Science Foundation (Priority Program for Biotechnology) and by an interdepartmental grant of the Swiss Federal Institute of Technology Lausanne (EPFL, Project Microtechnique 96). PMID- 11028059 TI - A Room-Temperature Discotic Nematic Liquid Crystal We are very grateful to Professor S. Chandrasekhar for helpful discussions. PMID- 11028061 TI - Multicomponent Reactions with Isocyanides. AB - Multicomponent reactions (MCRs) are fundamentally different from two-component reactions in several aspects. Among the MCRs, those with isocyanides have developed into popular organic-chemical reactions in the pharmaceutical industry for the preparation of compound libraries of low-molecular druglike compounds. With a small set of starting materials, very large libraries can be built up within a short time, which can then be used for research on medicinal substances. Due to the intensive research of the last few years, many new backbone types have become accessible. MCRs are also increasingly being employed in the total synthesis of natural products. MCRs and especially MCRs with isocyanides offer many opportunities to attain new reactions and basic structures. However, this requires that the chemist learns the "language" of MCRs, something that this review wishes to stimulate. PMID- 11028062 TI - Force Spectroscopy of Molecular Systems-Single Molecule Spectroscopy of Polymers and Biomolecules. AB - How do molecules interact with each other? What happens if a neurotransmitter binds to a ligand-operated ion channel? How do antibodies recognize their antigens? Molecular recognition events play a pivotal role in nature: in enzymatic catalysis and during the replication and transcription of the genome; it is also important for the cohesion of cellular structures and in numerous metabolic reactions that molecules interact with each other in a specific manner. Conventional methods such as calorimetry provide very precise values of binding enthalpies; these are, however, average values obtained from a large ensemble of molecules without knowledge of the dynamics of the molecular recognition event. Which forces occur when a single molecular couple meets and forms a bond? Since the development of the scanning force microscope and force spectroscopy a couple of years ago, tools have now become available for measuring the forces between interfaces with high precision-starting from colloidal forces to the interaction of single molecules. The manipulation of individual molecules using force spectroscopy is also possible. In this way, the mechanical properties on a molecular scale are measurable. The study of single molecules is not an exclusive domain of force spectroscopy; it can also be performed with a surface force apparatus, laser tweezers, or the micropipette technique. Regardless of these techniques, force spectroscopy has been proven as an extraordinary versatile tool. The intention of this review article is to present a critical evaluation of the actual development of static force spectroscopy. The article mainly focuses on experiments dealing with inter- and intramolecular forces-starting with "simple" electrostatic forces, then ligand-receptor systems, and finally the stretching of individual molecules. PMID- 11028063 TI - Does CH(5)(+) Have (a) "Structure?" A Tough Test for Experiment and Theory The author thanks Prof. Dr. W. Kutzelnigg for insightful comments and Prof. Dr. T. Oka for supplying original infrared data of CH(5)(+). PMID- 11028064 TI - Organometallic Transformations Demonstrate That Fluorocarbons Are Reactive Molecules. PMID- 11028065 TI - Calcium-Dependent Carbohydrate-Carbohydrate Recognition between Lewis(X) Blood Group Antigens This research was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie. PMID- 11028066 TI - Solid-Phase Synthesis of Doubly Labeled Peptide Nucleic Acids as Probes for the Real-Time Detection of Hybridization This work was supported by the Fonds der Chemischen Industrie. PMID- 11028067 TI - Redox Switches with Chiroptical Signal Expression Based on Binaphthyl Boron Dipyrromethene Conjugates This work was supported by a PhD fellowship from the Universiat Regensburg. We thank Prof. O. Wolfbeis for making the CD-spectrometer available. PMID- 11028068 TI - Allosteric Regulation of Artificial Phosphoesterase Activity by Metal Ions This work was funded by the DFG (Gerhard Hess Programm). PMID- 11028069 TI - Hexacyclinic acid, a Polyketide from Streptomyces with a Novel Carbon Skeleton We express our thanks to Hoechst AG (Frankfurt) for providing us with Streptomyces cellulosae (strain S 1013) and Mr. M. Decke for excellent technical assistance. We wish to thank Prof. Dr. W. Beil for determining the cytotoxic activity. This work was supported by the Fonds der Chemischen Industrie. PMID- 11028070 TI - The Dimerization of Chiral Allenes: Pairs of Enantiomers and Pairs of Homomers Furnish Different Diastereomers Cycloallenes, Part 14. This work was supported by the Deutsche Forschungsgemeinschaft, by the Fonds der Chemischen Industrie, and by CHEMETALL GmbH. Part 13: M. Christl, S. Groetsch, Eur. J. Org. Chem. 2000, 1871-1874. PMID- 11028071 TI - Five-Coordinate Carbides in Ti-Al-C Complexes Financial support from NSERC of Canada and Nova Chemicals Corporation is gratefully acknowledged. F.G. is grateful for the award of an NSERC Postdoctoral Fellowship. PMID- 11028072 TI - The First Example of an Equilibrium between a Carbene and an Isomeric Carbyne Transition Metal Complex This work was supported by the European Commission under the Training and Mobility of Researchers program (Marie Curie Fellowship to P. G. H., contract No. ERBFMBICT972450), the Deutsche Forschungsgemeinschaft (Grant SFB 347), and the Fonds der Chemischen Industrie (doctoral grant for K. I.). PMID- 11028073 TI - Model Studies of Phytochrome Photochromism: Protein-Mediated Photoisomerization of a Linear Tetrapyrrole in the Absence of Covalent Bonding This work is part of the PhD thesis of I. Lindner, Max-Planck-Institut fur Strahlenchemie, Mulheim an der Ruhr, and Gerhard-Mercator-Universitat, Duisburg, 2000. We thank Tanja Berndsen, Gul Koc, and Helene Steffen for technical assistance. PMID- 11028074 TI - C(60)F(18), a Flattened Fullerene: Alias a Hexa-Substituted Benzene This work was supported by the Royal Society, INTAS, the Russian Foundation for Fundamental Research (grant no. 99-03-32810), and the Russian Programme, "Fullerenes and Atomic Clusters". PMID- 11028075 TI - The First Microporous Framework Cerium Silicate This work was supported by PRAXIS XXI, FEDER, and FCT. PMID- 11028076 TI - Is Hydrogen Tunneling Involved in AcylCoA Desaturase Reactions? The Case of a Delta(9) Desaturase That Transforms (E)-11-Tetradecenoic Acid into (Z,E)-9,11 Tetradecadienoic Acid This work was supported by Comision Asesora de Investigacion Cientifica y Tecnica (grant AGF 98-0844), Comissionat per a Universitats i Recerca from the Generalitat de Catalunya (grant 97SGR-0021) and SEDQ S.A. We thank Prof. Nigel S. Scrutton (University of Leicester, UK) and Dr. Francisco J. Sanchez-Baeza (IIQAB, Barcelona, Spain) for helpful discussions, Dr. Josefina Casas and Dr. Antonio Delgado for critically reading the manuscript, and German Lazaro for rearing the insects used in this study. J.L.A. thanks the Spanish Ministry of Education and Science for a Postdoctoral Reincorporation Contract. PMID- 11028077 TI - Molecular Recognition with Introverted Functionality We are grateful for financial support from the Skaggs Research Foundation and the National Institutes of Health. We thank Prof. Dmitry Rudkevich and Dr. Paul Wash and Dr. Laura Pasternack for helpful advice and instrumental support. PMID- 11028078 TI - Towards Synthetic Molecular Muscles: Contraction and Stretching of a Linear Rotaxane Dimer We are grateful to M. Jean-Daniel Sauer and Dr. Roland Graff for high-field NMR experiments, Helene Nierengarten and Raymond Hubert for mass determinations. M.C.J. thanks the European Commission for a Grant (TMR Contract No. ERBFMBICT972547). We also thank the CNRS for financial support (Programme Physique et Chimie du Vivant). PMID- 11028079 TI - Conformation Control of Oligosilanes Based on Configurationally Constrained Bicyclic Disilane Units Financial support by a Grant-in-Aid from the Ministry of Education, Science, Sports, and Culture of Japan (09239103) is gratefully acknowledged. We thank Prof. R. West and Prof. J. Michl for their helpful discussions to this work. PMID- 11028080 TI - Asymmetric Synthesis of Baylis-Hillman-Type Allyl Alcohols via a Chiral Acetylenic Ester Titanium Alkoxide Complex We thank the Ministry of Education, Science, Sports and Culture (Japan) for financial support. PMID- 11028081 TI - Analysis of Solid-Phase Reactions: Product Identification and Quantification by Use of UV-Chromophore-Containing Dual-Linker Analytical Constructs We thank Dr. P. Marshall for the provision of analytical data. PMID- 11028082 TI - Redox-Switched Control of Binding Strength in Hydrogen-Bonded Metallocene Complexes. PMID- 11028083 TI - Toward Crystalline Covalent Solids: Crystal-to-Crystal Dihydrogen to Covalent Bonding Transformation in NaBH(4) small middle dot THEC The support of the MSU Center for Fundamental Materials Research is gratefully acknowledged. THEC=N,N',N",N"'-tetrakis-(2-hydroxyethyl)cyclen. PMID- 11028084 TI - NMR and Theoretical Study of Acid Sites Formed by Adsorption of SO(3) onto Oxide Surfaces This work was supported by the US NSF. PMID- 11028085 TI - Syntheses, Structures, and Magnetic Properties of Two Gadolinium(III)-Copper(II) Coordination Polymers by a Hydrothermal Reaction This work is supported by the NNSF of China, the NSF of Fujian Province, and the Key Project from the CAS. PMID- 11028086 TI - Experimental Observation of Pentaatomic Tetracoordinate Planar Si- and Ge Containing Molecules: MAl(4)(-) and MAl(4) The theoretical work was done at Utah State University and supported by the donors of The Petroleum Research Fund (ACS PRF no. 35255-AC6), administered by the American Chemical Society. The experimental work was supported by the National Science Foundation (DMR-9622733) and was performed at the W. R. Wiley Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by DOE's Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory, which is operated for DOE by Battelle Memorial Institute. L.S.W. is an Alfred P. Sloan Foundation Research Fellow. M=Si, Ge. PMID- 11028087 TI - Control of Architecture in Block-Copolymer Vesicles We thank the Petroleum Research Fund, administered by the American Chemical Society, for the support of this work. PMID- 11028088 TI - Hydrogen-Bonded Hexamolybdenum Clusters: Formation of Inorganic-Organic Networks This work was supported by the National Science Foundation (grant CHE-9417250). PMID- 11028089 TI - A Combinatorial Approach to Polyketide-Type Libraries by Iterative Asymmetric Aldol Reactions Performed on Solid Support We thank the European Commission (TMR Network ERB-FMR XCT 96-0011 and IHP Network HPRN-CT-2000-00014), EPSRC, Pfizer, and Merck for support. PMID- 11028090 TI - Synthesis, Structure, and Reactivity of a 1sigma(4),3sigma(2)-Diphosphaallene We are grateful to the French Embassy in Japan for a grant to T.K., and to the CNRS for financial support of this work. PMID- 11028091 TI - 4,5-Bis(diphenylphosphinoyl)-1,2,3-triazole: A Powerful New Ligand That Uses Two Different Modes of Chelation. PMID- 11028092 TI - In memoriam carlos chagas filho PMID- 11028093 TI - Carlos chagas filho, scientist and citizen PMID- 11028094 TI - Bernstein-type theorems in hypersurfaces with constant mean curvature AB - By using the nodal domains of some natural function arising in the study of hypersurfaces with constant mean curvature we obtain some Bernstein-type theorems. PMID- 11028095 TI - [ [In Process Citation] AB - The author gives a brief description of the studies that led to the eclosion of the weak forces model of unification of Electromagnetic interactions with weak interactions. PMID- 11028096 TI - Surface charges and interfaces: implications for mineral roles in prebiotic chemistry AB - There exists an extensive literature on the possible roles of minerals in the prebiotic stages of the chemical evolution of life (Bernal 1951, Cairns-Smith 1982, Wachtershauser 1992, Vieyra et al. 1995, Tessis et al. 1999, see Lahav (1994) for a review). Among the original proposals, minerals have been considered in: (a) processes that would discriminate molecular chirality; (b) condensation reactions of biomolecular precursors; (c) prebiotic catalysis; (d) biochemical templates; and (e) autocatalytic metabolism. In this communication it is emphazised the complex properties of both surface reactions and interfaces between minerals and aqueous solutions simulating Archean scenarios. The properties of pyrite surface net charge and of its interface with a solution simulating primitive seawater are discussed and their implications to the autocatalytic model (Wachtershauser 1988a 1992) are presented in order to demonstrate their relevance. The proposed roles of iron-sulfide minerals (mainly pyrite) as physical support for primitive bidimensional metabolism and chiral discriminator (Wachtershauser 1988a, Huber & Wachtershauser 1998) are revised. It is shown that: (a) the net surface charge can be modulated by the pyrite-aqueous solution interface; (b) mononucleotides attachment to pyrite require a cationic bridge; and (c) direct absorption of acetate - a molecule proposed as carbon source in primitive aqueous environments - also modulates the interface properties and would have masked pyrite's bulk structure. These results indicate that physicochemical changes of mineral surfaces - caused by environments simulating Archean aqueous scenarios - should be taken into account in the proposals of mineral prebiotic roles. PMID- 11028097 TI - Bird community as an indicator of biodiversity: results from quantitative surveys in Brazil. AB - This short review presents the results obtained in several localities of Brazil on the composition of forest bird communities. Data were collected since the late 80's, after we introduced a new methodology of quantitative survey, based on acoustic identification and unlimited-radius point census. Although these data are still scattered, they show uniquely precise and coherently comparative patterns of composition of forest bird communities. Our methodology has the advantage of being absolutely non-disturbing, highly efficient in the field and immediately processed. Results confirm that the structure of a bird community is a good indicator of biodiversity, particularly useful where biodiversity is high. Many of these data are available only in unpublished dissertations and abstracts of congress communications, or are being analysed. A cooperative program is needed to promote new surveys and publish their results, as a contribution for measuring and monitoring biodiversity, especially in complex endangered habitats. PMID- 11028098 TI - Choline acetyltransferase detection in normal and denervated electrocyte from Electrophorus electricus (L.) using a confocal scanning optical microscopy analysis. AB - Acetylcholine is the neurotransmitter responsible for the transmission of impulses from cholinergic neurons to cells of innervated tissues. Its biosynthesis is catalyzed by the enzyme Choline acetyltransferase that is considered to be a phenotypically specific marker for cholinergic system. It is well known that the regulation of Choline acetyltransferase activity under physiological and pathological conditions is important for development and neuronal activities of cholinergic functions. We observed the distribution of Choline acetyltransferase in sections from the normal and denervated main electric organ sections of Electrophorus electricus (L.) by immunofluorescence using a anti-Choline acetyltransferase antibody. The animals were submitted to a surgical procedure to remove about 20 nerves and after 30 and 60 days, they were sacrificed. After 30 days, the results from immunohistochemistry demonstrated an increase on the Choline acetyltransferase distribution at denervated tissue sections when compared with the sections from the normal contralateral organ. A very similar labeling was observed between normal and denervated tissue sections of the animals after 60 days. However, Choline acetyltransferase activity (nmolesACh/ min/ mg of protein) in extracts obtained from electrocyte microsomal preparation, estimated by Fonnun's method (Fonnun 1975), was 70% lower in the denervated extracts. PMID- 11028099 TI - The cytoskeleton of the electric tissue of Electrophorus electricus, L. AB - The electric eel Electrophorus electricus is a fresh water teleost showing an electrogenic tissue that produces electric discharges. This electrogenic tissue is distributed in three well-defined electric organs which may be found symmetrically along both sides of the eel. These electric organs develop from muscle and exhibit several biochemical properties and morphological features of the muscle sarcolema. This review examines the contribution of the cytoskeletal meshwork to the maintenance of the polarized organization of the electrocyte, the cell that contains all electric properties of each electric organ. The cytoskeletal filaments display an important role in the establishment and maintenance of the highly specialized membrane model system of the electrocyte. As a muscular tissue, these electric organs expresses actin and desmin. The studies that characterized these cytoskeletal proteins and their implications on the electrophysiology of the electric tissues are revisited. PMID- 11028100 TI - Short- and long-term memory: differential involvement of neurotransmitter systems and signal transduction cascades. AB - Since William James (1890) first distinguished primary from secondary memory, equivalent to short- and long-term memory, respectively, it has been assumed that short-term memory processes are in charge of cognition while long-term memory is being consolidated. From those days a major question has been whether short-term memory is merely a initial phase of long-term memory, or a separate phenomena. Recent experiments have shown that many treatments with specific molecular actions given into the hippocampus and related brain areas after one-trial avoidance learning can effectively cancel short-term memory without affecting long-term memory formation. This shows that short-term memory and long-term memory involve separate mechanisms and are independently processed. Other treatments, however, influence both memory types similarly, suggesting links between both at the receptor and at the post-receptor level, which should not be surprising as they both deal with nearly the same sensorimotor representations. This review examines recent advances in short- and long-term memory mechanisms based on the effect of intra-hippocampal infusion of drugs acting upon neurotransmitter and signal transduction systems on both memory types. PMID- 11028101 TI - Energy interconversion by the sarcoplasmic reticulum Ca2+-ATPase: ATP hydrolysis, Ca2+ transport, ATP synthesis and heat production. AB - The sarcoplasmic reticulum of skeletal muscle retains a membrane bound Ca2+ ATPase which is able to interconvert different forms of energy. A part of the chemical energy released during ATP hydrolysis is converted into heat and in the bibliography it is assumed that the amount of heat produced during the hydrolysis of an ATP molecule is always the same, as if the energy released during ATP cleavage were divided in two non-interchangeable parts: one would be converted into heat, and the other used for Ca2+ transport. Data obtained in our laboratory during the past three years indicate that the amount of heat released during the hydrolysis of ATP may vary between 7 and 32 Kcal/mol depending on whether or not a transmembrane Ca2+ gradient is formed across the sarcoplasmic reticulum membrane. Drugs such as heparin and dimethyl sulfoxide are able to modify the fraction of the chemical energy released during ATP hydrolysis which is used for Ca2+ transport and the fraction which is dissipated in the surrounding medium as heat. PMID- 11028102 TI - Ubiquitous overexpression of a transgene encoding the extracellular portion of the Drosophila roughest-irregular chiasm C protein induces early embryonic lethality. AB - The cell adhesion molecule Rst-irreC is a transmembrane glycoprotein of the immunoglobulin superfamily involved in several important developmental processes in Drosophila, including axonal pathfinding in the optic lobe and programmed cell death and pigment cell differentiation in the pupal retina. As an initial step towards the "in vivo" functional analysis of this protein we have generated transgenic fly stocks carrying a truncated cDNA construct encoding only the extracellular domain of Rst-IrreC under the transcriptional control of the heat shock inducible promoter hsp70. We show that heat-shocking embryos bearing the transgene during the first 8hs of development lead to a 3-4 fold reduction in their viability compared to wild type controls. The embryonic lethality can already be produced by applying the heat pulse in the first 3hs of embryonic development, does not seem to be suppressed in the absence of wildtype product and is progressively reduced as the heat treatment is applied later in embryogenesis. These results are compatible with the hypothesis of the lethal phenotype being primarily due to heterophilic interactions between Rst-IrreC extracellular domain and an yet unknown ligand. PMID- 11028103 TI - Genetic modification of mammalian genome at chromosome level. AB - The review is concerned with a progress in genetic modification of a mammalian genome in vitro and in vivo at chromosomal level. Recently three new approaches for the chromosome biotechnology have been developed: Using Cre/loxP-system a researcher is able to produce targeted rearrangements of whole chromosomes or their segments or particular genes within the genome, and therefore to modify the set, position and copy number of the endogenous elements of the genome. Mammalian artificial chromosomes (MACs) provide a possibility to introduce into genome relatively large segments of alien chromosome material, either artificially constructed or derived from the genome of different species. Using ES-somatic cell hybrids allows to transfer whole chromosomes or their fragments between different genomes within and between species. Advantages and limitations of these approaches are discussed. PMID- 11028104 TI - The cystic fibrosis transmembrane regulator (CFTR) in the kidney. AB - The cystic fibrosis transmembrane regulator (CFTR) is a Cl - channel. Mutations of this transporter lead to a defect of chloride secretion by epithelial cells causing the Cystic Fibrosis disease (CF). In spite of the high expression of CFTR in the kidney, patients with CF do not show major renal dysfunction, but it is known that both the urinary excretion of drugs and the renal capacity to concentrate and dilute urine is deficient. CFTR mRNA is expressed in all nephron segments and its protein is involved with chloride secretion in the distal tubule, and the principal cells of the cortical (CCD) and medullary (IMCD) collecting ducts. Several studies have demonstrated that CFTR does not only transport Cl - but also secretes ATP and, thus, controls other conductances such as Na+ (ENaC) and K+ (ROMK2) channels, especially in CCD. In the polycystic kidney the secretion of chloride through CFTR contributes to the cyst enlargement. This review is focused on the role of CFTR in the kidney and the implications of extracellular volume regulators, such as hormones, on its function and expression. PMID- 11028105 TI - Sodium pumps in the Malpighian tubule of Rhodnius sp. AB - Malpighian tubule of Rhodnius sp. express two sodium pumps: the classical ouabain sensitive (Na+ + K+)ATPase and an ouabain-insensitive, furosemide-sensitive Na+ ATPase. In insects, 5-hydroxitryptamine is a diuretic hormone released during meals. It inhibits the (Na+ + K+)ATPase and Na+ -ATPase activities indicating that these enzymes are involved in fluid secretion. Furthermore, in Rhodnius neglectus, proximal cells of Malpighian tubule exposed to hyperosmotic medium, regulate their volume through a mechanism called regulatory volume increase. This regulatory response involves inhibition of the (Na+ + K+)ATPase activity that could lead to accumulation of active osmotic solute inside the cell, influx of water and return to the normal cell volume. Adenosine, a compound produced in stress conditions, also inhibits the (Na+ + K+)ATPase activity. Taken together these data indicate that (Na+ + K+)ATPase is a target of the regulatory mechanisms of water and ions transport responsible for homeostasis in Rhodnius sp. PMID- 11028106 TI - Inositol metabolism in Trypanosoma cruzi: potential target for chemotherapy against Chagas' disease. AB - Chagas' disease is a debilitating and often fatal disease caused by the protozoan parasite Trypanosoma cruzi. The great majority of surface molecules in trypanosomes are either inositol-containing phospholipids or glycoproteins that are anchored into the plasma membrane by glycosylphosphatidylinositol anchors. The polyalcohol myo-inositol is the precursor for the biosynthesis of these molecules. In this brief review, recent findings on some aspects of the molecular and cellular fate of inositol in T. cruzi life cycle are discussed and identified some points that could be targets for the development of parasite-specific therapeutic agents. PMID- 11028107 TI - Two special organelles found in Trypanosoma cruzi. AB - We review here two unique organelles from Trypanosoma cruzi. One of them is the acidocalcisome, cytoplasmic vacuoles containing a very high Ca2+ concentration and a Ca2+ - H+ translocating ATPase activity, present in all trypanosomatids. The other organelle is the reservosome, site of accumulation of endocytosed macromolecules, very rich in cysteine proteinase, that is present only in epimastigote forms of trypanosomes belonging to the Schyzotrypanum sub-genus. PMID- 11028108 TI - A static paradigm (and/or) a dynamic thought. PMID- 11028109 TI - Markers for vitiligo related neuropeptides in human skin nerve fibers. AB - Skin distribution of substance P (SP)-, somatostatin (SOM)-, calcitonin-gene related peptide (CGRP)- and neuropeptide-Y (NPY)-like immunoreactivity in vitiligo patients was studied by an indirect immunofluorescence technique. Immunocytochemical characteristics of the epidermis, dermoepidermal junction, papillary and reticular dermis, and skin appendages were analyzed in lesional and marginal vitiligo areas as well as in healthy skin. SP-, SOM-, CGRP-, and NPY immunoreactive nerve fibers were observed in healthy pigmented skin, with patterns specific for immunoreactive distribution. Thin SP-containing fibers were observed in dermal papillae, extending into the epidermis, and SP-immunoreactive nerve fibers were seen around blood vessels and sweat glands. SOM-immunoreactive varicose nerve fibers were associated with Meissner's corpuscles in dermal papillae, while CGRP-like immunoreactivity was demonstrated in free subepidermal nerve terminals and sensory nerve fibers around blood vessels, hair follicles and sweat glands. Autonomic NPY-containing nerve fibers innervated eccrine sweat glands and blood vessels. The distribution of these neuropeptides was the same in healthy controls, except for an increased immunoreactivity to NPY and to a lesser extent to CGRP. These results suggest that NPY may serve as a neurochemical marker in the pathogenesis of the disease, thus supporting the neuronal theory of vitiligo. PMID- 11028110 TI - Comparative immunohistochemical analysis of estrogen receptor and chromogranin-A reactivity in plurihormonal human prolactinomas. AB - The aim of the study was to compare the immunoreactivity of estrogen receptors (ER) and chromogranin-A (CHR-A) in human prolactinomas with verified plurihormonality. Eleven cases of prolactinomas, nine found in women aged from 15 32 and two found in two men both aged 54 years, were analyzed for possible colocalization of other hormones produced by adenohypophysis, i.e. growth hormone (GH), thyroid-stimulating hormone (TSH), follicle-stimulating hormone (FSH) and adrenocorticotropic hormone (ACTH). All evaluated cases of prolactinomas were clinically manifested by elevated values of prolactin (PRL) in patient serum, while the values of other assayed hormones were within the normal range. Although biopsy material is not routinely submitted to immunohistochemical analysis for plurihormonality, these eleven cases of operated prolactinomas were randomly examined to the presence of plurihormonality. In six cases of prolactin-producing adenomas, the coexistence of growth hormone was detected. Colocalization of follicle-stimulating hormone and weak expression of adrenocorticotropic hormone were found in two cases each. Thus, bihormonal activity (PRL + GH) was found in six, and trihormonal activity (PRL + GH + FSH and PRL/GH + ACTH) in three cases of prolactinoma. In addition, the presence of prolactin and growth hormone was demonstrated in morphologically different cells. Eight of these eleven pituitary adenomas were tested for estrogen receptors (ER), which play an important role as growth stimulating factors and secretory factors for prolactin-producing cells. We tried to determine if there was a difference in the intensity of expression of estrogen receptors and chromogranin-A between pure prolactinomas and mixed, plurihormonal prolactinomas. By use of monoclonal antibodies, chromogranin-A found to be reactive in seven of eleven prolactinomas, i.e. in plurihormonal prolactinomas. Estrogen receptors were markedly expressed in all the eight prolactinomas analyzed, which may prove significant in the treatment of these hypophyseal tumors. PMID- 11028111 TI - Pilocarpine in the prevention of postirradiation xerostomia. AB - During radiation therapy to the head and neck region, salivary gland hypofunction commonly develops. The aim of our study was to evaluate whether the sialogogue pilocarpine given during radiation therapy may reduce the severity of xerostomia and salivary dysfunction. Our results showed the pilocarpine-treated group to have a lower frequency of oral symptoms during the treatment than the placebo treated group. Although salivary flow decreased in all patients, the pilocarpine treated group had smaller flow reductions. No drug effect was observed in the glands that were completely irradiated. Thus, pilocarpine appeared to stimulate salivary tissues outside the radiation field. PMID- 11028112 TI - Slow continuous renal replacement therapies: an update. AB - Continuous renal replacement therapies (CRRT) are now being used by nephrologists, intensivists, and anesthesiologists. The various CRRT modalities differ in the kind of vascular access, the application of diffusive or convective clearances (or a combination of both), and in the location where the replacement fluid enters the circuit. CRRTs have certainly made the management of critically ill patients with acute renal failure (ARF) combined with cardiovascular instability, severe fluid overload, hypercatabolism, cerebral edema, adult respiratory distress syndrome, lactic acidosis, sepsis or other inflammatory syndromes, crush syndrome, congestive heart failure, and cardiopulmonary bypass easier. Continuous therapies incorporate several advantages including improved hemodynamic stability, optimal fluid balance, gradual urea removal, elimination of septic mediators, and the possibility of unlimited parenteral nutrition. Major difficulties and unsolved problems of CRRT are the ongoing necessity of continuous anticoagulation, considerable loss of amino acids, vitamins, trace elements, potassium, phosphate, and some drugs, as well as immobilization of the patient. The advantages of CRRT should theoretically translate into improved outcomes of critically ill ARF patients, but the superiority of continuous modalities in terms of outcome is still controversial, despite encouraging results in some clinical trials. Currently used CRRT with sophisticated treatment devices has become more expensive than hemodialysis, but the cost cannot be used as an argument against the continuous treatment modalities. PMID- 11028113 TI - Recruitment in the diagnosis of small PCA tumors. AB - Acoustic brainstem response (ABR) waveforms were preoperatively analyzed in 86 cases of pontocerebellar angle (PCA) tumors. Absolute (APL) and interpeak latencies (IPL) ipsilateral to the tumor and interaural absolute latency differences of wave V (IAAPLD V) were normal in two patients with small intracanalicular tumors. However, results of latency-intensity studies were abnormal. So, if PCA tumor is suspected, and standard ABRs are normal by usual criteria, the latency-intensity functions should be examined before the test result is reported as negative for an acoustic nerve tumor. PMID- 11028114 TI - On the occasion of the 45th anniversary of the discovery of lysosomes. PMID- 11028115 TI - Guillian Barre syndrome--recent advances. AB - Guillian Barre Syndrome (GBS) is an acquired disease of the peripheral nerves that is characterized clinically by rapidly progressing paralysis, areflexia, and albumino-cytological dissociation. It affects both genders, involves people of all ages, is reported worldwide, and in the post-polio era, it is the most common cause of an acute generalized paralysis. The clinical features are distinct and a history and an examination generally lead to a high suspicion of the diagnosis that can then be confirmed by supportive laboratory tests and electrodiagnostic studies. This review discusses the recent advances in understanding of the different variants of GBS such as acute inflammatory demyelinating polyneuropathy (AIDP), acute motor axonal neuropathy (AMAN), acute motor sensory axonal neuropathy (AMSAN), and the Fisher syndrome. The clinical, electrodiagnostic criteria, immunopathogenesis, and management of GBS and its variants are discussed. PMID- 11028116 TI - Evaluation of pulse-polio immunisation in rural area of Maharashtra. AB - An evaluation survey on Pulse-polio Immunisation (PPI) in January 2000 was carried out in rural areas of Maharashtra to assess the immunisation coverage, the knowledge regarding pulse-polio and the routine immunisation schedule. House to-house survey was carried out and information was collected by interviewing the parents of 778 children. It was observed that PPI coverage was excellent i.e., 98%. Majority of informants were female. Knowledge about pulse-polio had a direct relationship with literacy. But knowledge regarding routine immunisation and the polio disease was not found satisfactory. Excellent coverage of pulse-polio Immunisation was found because of an organised and extensive campaign, use of mass media like T.V., radio, and home visits of peripheral health staff (as told by the informants). The distance of approach to a polio booth was less than a kilometre in almost all cases. Behaviour of health staff was satisfactory everywhere. The time-period required for getting vaccinated after arrival at the polio booth was less than 5 minutes. Thus it was a worthwhile attempt to evaluate PPI coverage is rural areas of Maharasthra. Excellent coverage of pulse-polio immunisation gives us the picture that poliomyelitis is on the verge of being eradicated from India, if the excellent coverage of PPI is followed by effective surveillence. PMID- 11028117 TI - Nasal mucociliary clearance in adenotonsillar hypertrophy. AB - The nasal mucociliary clearance time was studied using Andersen saccharin method in 50 normal children and 50 age and sex matched patients of adenotonsillar hypertrophy, which was repeated one month after adenotonsillectomy. The normal mucociliary clearance time in healthy children was found to be 8.55 +/- 2.11 minutes. A significant impairment in nasal mucociliary clearance time was noted in children suffering with adenotonsillar hypertrophy which was 16.97 +/- 3.1 minutes, and early adenotonsillectomy restored the mucociliary clearance to a normal 8.7 +/- 2.14 minutes. PMID- 11028118 TI - Hyperthyroidism in children. AB - This study was done to characterize the clinical features, laboratory parameters and response to therapy and outcome of childhood hyperthyroidism. The evaluation included history, examination, laboratory investigations: serum T3, T4, TSH, free T3, free T4 by RIA or immunochemiluminescence (IC), antithyroid antibodies by standard techniques, bone age (BA) by Greulich and Pyle's method, clinical and laboratory response to treatment, and follow-up of 15 children with hyperthyroidism seen in past eight years. Age of onset, presentation, nature and duration of symptoms, family history, anthropometry and signs of hyperthyroidism were recorded. There were 10 girls and 5 boys (2:1). Three families had a history of thyroid disorders. Mean ages of onset and presentation were 8.25 +/- 3.4 and 9.27 +/- 3.2 years respectively. Clinical features included weight loss, heat intolerance and sweating, diarrhoea, behavioral problems, ophthalmopathy and tachycardia. BA was advanced. Serum T3 (mean = 4.29 +/- 1.77 ng/mL), T4 (18.75 +/ 5.64 micrograms/dL), FT3 (7.11 +/- 4.58 pg/mL) and FT4 (2.93 +/- 0.29 ng/mL) were markedly elevated. TSH was suppressed. Anti-microsomal antibodies (AMA) and anti-thyroglobulin antibodies (ATG) were positive in five. They were started on standard treatment with carbimazole 0.5-0.7 mg kg-1. Clinical and biochemical euthyroidism was achieved within 2.5 to 6 months in all, after which the drug was tapered, however, they required treatment for 2 years to 7.5 years. Four children were retreated for relapse and are now euthyroid and off treatment. Childhood hyperthyroidism requires long term treatment and careful monitoring. This study shows a remission rate of 67%. PMID- 11028119 TI - Choledochal cyst: a 10-year experience. AB - A retrospective study of 11 cases of choledochal cyst over a period of 10 years is presented. There were 8 females and 3 males in a ratio of 2.67:1. The classical triad of jaundice, mass and abdominal pain was seen in none of our patients. Ultrasonography was diagnostic in 9 out of 11 (80%) patients. Surgical procedures performed were, complete excision of the cyst with Roux en Y hepaticojejunostomy (nine cases), internal drainage of the cyst (one case) and excision of the extrahepatic cyst with Roux en Y hepaticojejunostomy in one case of Type IV choledochal cyst. One patient died in the immediate postoperative period. Others have remained well upto 10 years follow-up. We recommend total cyst excision with hepaticojejunostomy as the treatment of choice. PMID- 11028120 TI - Isolated congenital coronary arteriovenous fistula. AB - During the last eight years a total of nine cases of isolated congenital coronary arteriovenous fistula (CAVF) have been diagnosed at our centre. Age of these patients ranged from 11 days to 22 years (mean 6.4 +/- 4.2 years). There were seven male and two female patients. Right and left coronary arteries were the sites of origin in seven and three cases respectively. One patient had two fistulae, one each originating from right coronary artery and obtuse marginal artery. The drainage sites were right ventricle in seven cases and right atrium in two. The pulmonary artery pressure was normal in eight cases and elevated in one case. The pulmonary artery to systemic flow ratio ranged from 1.1:1-2.4:1 in all except the 11 day-old-baby in whom it was 4:1. Pulmonary vascular resistance (PVRI) was below 3.5 in all cases (range 1.0-3.5 units). Surgical ligation of fistula was done in four cases and two patients underwent coil embolisation of fistula. Follow-up is available in five out of eight patients including three cases in whom CAVF was closed. Two other young patients are being followed-up conservatively and all the patients are asymptomatic. CAVF can present at any age; majority of the cases have small left to right shunts and more commonly CAVF arise from right coronary artery. Surgical ligation of fistula is safe in older patients, non-surgical embolisation using coils may be an alternative to surgery in selected cases. PMID- 11028122 TI - Intestinal atresia. AB - Intestinal atresia accounts for about one third of all cases of neonatal intestinal obstruction. The survival rate has improved to 90% in most of the series with the operative mortality being < 1%. The survival rate improves with distal atresias. An increased mortality is observed in multiple atresias (57%), apple peel atresia (71%), and when atresia is associated with meconium ileus (65%), meconium peritonitis (50%) and gastroschisis (66%). Although appearance of echogenic bowel on prenatal ultrasonography is suggestive of GI, it is confirmed in only 27% cases. Prenatal ultrasonography is more reliable in detection of duodenal atresia than more distal lesions. Short bowel syndrome is the major impediment in the management of jejunoileal atresia. Although total parenteral nutrition (TPN) is the main adjunctive treatment, it delays intestinal adaptation and may cause cholestasia and subsequent liver damage. Graduated enteric feedings, use of growth hormone, glutamine and modified diets containing low fat, complex carbohydrates and protein supplements have been used in a adults with short bowel syndrome to successfully diminish TPN requirements and enhance nutrient absorption in nearly half of the patients. Utilization of growth factors to facilitate intestinal adaptation and advances in small bowel transplant may improve the long-term outcomes in future. PMID- 11028121 TI - Congenital diaphragmatic hernia. AB - Over the last two decades there has been a constant improvement in the understanding of the pathophysiology of Congenital Diaphragmatic Hernia (CDH) and its management. However, the ideal treatment remains elusive. The earlier management strategy of immediate surgery is replaced by the principle of physiological stabilisation and delayed surgery. Conventional mechanical ventilatory techniques, with high pressures and hyperventilation to reverse ductal shunting and cause alkalinization, are being questioned because of the risks of barotrauma and consequent broncho-pulmonary dysplasia. It has also been shown that paralysis with pancuronium bromide for patients on conventional mechanical ventilation results in increased incidence of sensorineural hearing loss in childhood survivors of CDH. With the introduction of the concept of permissive hypercapnia and high frequency oscillation ventilation, the complications of pulmonary barotrauma are circumvented. Although ECMO therapy is invasive, yet has improved survival by about 15% independently, especially in critically ill infants who have the predictive mortality rate of more than 80%. Further insights into the pathophysiology of CDH and the introduction of less invasive therapeutic techniques in the form of high frequency oscillation ventilation, inhalation nitric oxide, surfactant, and perfluorocarbon liquid ventilation may even make the need for ECMO redundant. PMID- 11028123 TI - Renal amyloidosis following tuberculosis. AB - Amyloidosis, either primary or secondary, may be defined as a group of chronic infiltrative disorders that have in common a beta-pleated sheet configuration on X-ray diffraction examination, a fine fibrillar nonbranching appearance on electron microscopy and an apple-green birefringence when examined under polarised light after staining with Congo-red. Renal amyloidosis is a rare entity in the pediatric age group and is almost always secondary in nature, related to chronic infections and inflammatory conditions. It occurs 2-7 years after a chronic inflammatory process; however an onset as early as 9 months of life is known. The diagnosis of amyloidosis is suspected on the basis of clinical features and is established by obtaining an appropriate tissue biopsy and demonstrating amyloid with appropriate stains. All the tissues obtained must be stained with Congo-red stain which is the singlemost useful diagnostic test to define amyloidosis. In order to differentiate the primary from secondary variety, the deposits may be treated with potassium permanganate before Congo-red staining. In secondary amyloidosis, the green birefringence seen under polarized light is abolished. Therapeutic approaches include specific measures to reduce the amyloid deposition and general measures to relieve symptoms related to involvement of specific organs. The prognosis in renal amyloidosis is relatively poor, with a median survival of 9-13 months in primary amyloidosis complicated by renal involvement, and more than 50 months in secondary amyloidosis. We have reported a case of secondary amyloidosis following tuberculosis and have discussed the clinical features, diagnosis and management of amyloidosis. PMID- 11028124 TI - I-cell disease (Mucolipidosis II). AB - I-cell disease (Mucolipidosis II) is one of the lysosomal storage diseases which presents in the neonatal period, and within six months will phenotypically resemble the severe forms of the group of disorders called the "mucopolysaccharidoses" but without mucopolysacchariduria. In Mucolipidosis II, fibrocytes exhibit "abnormal lysosomes". Activities of several lysosomal enzymes are low in fibroblast cultures but high in mucolipidosis II serum. We present a patient with I-cell disease diagnosed on the basis of clinical, radiological and biochemical features. The mother of this child was pregnant and the fetus was also found to be affected. PMID- 11028125 TI - Juvenile neuronal ceroid lipofuscinosis. AB - A case of juvenile neuronal ceroid lipofuscinosis (JNCL) diagnosed on the basis of clinical features, electrophysiologic studies and skin electron microscopy is reported. JNCL was suspected on the basis of characteristic symptoms including progressive loss of vision, seizures, mental retardation and motor disabilities. Diagnosis was confirmed by neurophysiological and biopsy studies. The disease is caused by 23 different mutations in a gene recently isolated on chromosome 16 p11.2-12.1. Although universally fatal, characterisation of mutations can help in prenatal diagnosis in future pregnancies. PMID- 11028126 TI - Idiopathic pulmonary hemosiderosis. AB - Two cases of idiopathic pulmonary hemosiderosis in children are reported. Both cases presented with a combination of acute/recurrent respiratory symptoms along with iron deficiency anemia. In one case diagnosis was delayed for 18 months after onset of symptoms. After initial stabilisation with corticosteroid therapy, both cases were put on chloroquine therapy and showed improvement. Various other forms of therapy and outcome are discussed. It is suggested that in cases with recurrent respiratory symptoms, idiopathic pulmonary hemosiderosis should also be considered after excluding infectious etiology. PMID- 11028127 TI - Hepatobiliary abnormalities and parenteral nutrition. AB - Hepatobiliary dysfunctions (TPN-HBD) occur during parenteral nutrition. In older children these are usually reversible whereas in newborns and infants these hepatobiliary abnormalities play a significant role in the morbidity. Cholestasis is a commonly occurring TPN-HBD. It correlates directly with the decreasing gestational age, low birth weight and increasing duration of TPN therapy. The pathogenesis of cholestasis of TPN is multifactorial and predisposed by necrotising enterocolitis, sepsis, cardiac failure, shock, and hypotension. Diagnosis is made with exclusion of other causes of direct hyperbilirubinemia. Most TPN-HBD appear within 4 weeks of starting of TPN but severe complications manifest usually after the 16th week. Histologically there is intralobular cholestasis. In few cases there may be severe portal fibrosis followed by development of micronodular biliary cirrhosis. Enteral starvation, defective bile acid carriers, hypercaloric TPN are the major factors responsible for TPN-HBD, including cholestasis. Biliary complications of TPN-HBD are acalculous, cholecystitis, and cholelithiasis. Bile stasis is a major pathological factor for these. If the calories are provided only by glucose or glucose-containing electrolyte solutions it may lead to cholestasis and other TPN-HBD. Even small oral alimentation (continuous or bolus) during TPN, prevent TPN-HBD. Choleretic agents have been useful in the prevention and management of cholestasis and other parenteral nutrition induced hepatobiliary abnormalities. PMID- 11028128 TI - Comments on Jordans' anomaly. PMID- 11028129 TI - The U.K. NHS economic evaluation database. Economic issues in evaluations of health technology. AB - OBJECTIVE: The U.K. NHS Economic Evaluation Database (EED) project is commissioned to identify papers on economic evaluations of health technologies and to disseminate their findings to NHS decision makers by means of structured abstracts that are available through a public database and the Cochrane Library. This paper discusses current issues relating to the economic aspects of producing NHS EED abstracts. METHODS: A review of NHS EED was undertaken between 1994 and 1999 to determine the methodologies adopted and issues that influence the usefulness of economic evaluations. Methods adopted to improve the quality of NHS EED abstracts are also reported. RESULTS: Eighty-five percent of NHS EED abstracts are cost-effectiveness analyses (CEAs), 9.3% are cost-utility analyses (CUAs), and only 1.4% are cost-benefit analyses (CBAs). Of the total abstracts, 65.9% are based on single studies, 19.5% on reviews, 3.9% on estimates of effectiveness, and 10.7% on combinations of these sources. Models are utilized in 16.7% of CEAs, 60.2% of CUAs, and 20% of CBAs. Analyses of CBA studies reveal a degree of misuse of well-established definitions. NHS EED internal control mechanisms are reported that provide a means of ensuring that abstracts are based on sound academic principles. CONCLUSIONS: Most economic evaluations are conducted by means of CEA, followed by CUA, while CBA accounts for an extreme minority of cases. Single studies form the principal source of effectiveness data, although models are widely used, principally in CUA. The structure of NHS EED abstracts provides decision makers with the principal results and an interpretation of the relative strengths and weaknesses of economic evaluations. PMID- 11028130 TI - What are the relative merits of the sources used to identify potential research priorities for the NHS HTA programme? AB - The NHS Health Technology Assessment (HTA) Programme runs an annual process of identifying suggestions for health technology assessment. The objective of this paper is to describe and evaluate the relative importance of the different sources used by the program in 1998 to identify potential priorities. There were four different sources: a) a widespread consultation of healthcare commissioners, providers and consumers; b) research recommendations from systematic reviews; c) reconsidering previous research priorities which had not been taken forward for funding; and d) horizon scanning. Collectively, the four sources generated just over 1,100 HTA suggestions. By far the largest source of suggestions and priorities was the widespread consultation. However, the success rate of this source, in terms of being commissioned, was low. Research recommendations from systematic reviews provided the second largest source of priorities and the best success rate of all sources. Value was found from different sources for different healthcare areas. PMID- 11028131 TI - Cost-effectiveness of interferon beta-1b in slowing multiple sclerosis disability progression. First estimates. AB - OBJECTIVE: To estimate the cost-effectiveness (CE) of interferon beta-1b (IFN beta-1b) in slowing disability progression in persons with relapsing-remitting multiple sclerosis (RRMS). METHODS: Treatment program costs and health outcomes are modeled for cohorts of 1,000 females and 1,000 males followed 40 years from onset. Fifteen scenarios model MS natural history progression, treatment efficacy, direct treatment costs, and MS healthcare costs. A single randomized placebo-controlled trial of IFN beta-1b found reduced disease activity by MRI, reduced frequency and severity of exacerbations, and a tendency toward slower disability progression. Disability years avoided are modeled as the primary health outcome analyzed. A ministry of health (MOH) perspective is adopted, using Nova Scotia population-based data. Annual IFN beta-1b direct treatment costs (Can $16,685) are high relative to both MOH healthcare costs per person with MS (Can $2,000) and estimated MOH costs avoided. RESULTS: Given "reference case" assumptions for women with RRMS, treatment reduces lifetime disability years by 10%. Cost per disability year avoided before discounting is Can $189,230 (US $124,892), and Can $274,842 (US $181,395) after discounting at 5%. Estimates for alternative scenarios vary greatly, leaving main findings unchanged. CONCLUSIONS: Using the Expanded Disability Status Scale, cost per disability year avoided due to interferon beta-1b treatment in RRMS is quite high. Comparable CE estimates, using MS-specific or generic health-related quality-of-life outcome measures, are even higher. Further research is required to better measure treatment effects, modification of MS natural history, and net societal costs of IFN beta-1b in RRMS. PMID- 11028132 TI - Cost-utility analysis of interferon beta-1b in secondary progressive multiple sclerosis. AB - OBJECTIVES: Interferon beta-1b has recently become available for the treatment of secondary progressive multiple sclerosis (SPMS). This study aims at estimating the cost-effectiveness of this new treatment that has been shown in a clinical trial to reduce the progression of the disease. Effectiveness is measured as the number of quality-adjusted life-years (QALYs) gained from the reduction in progression. Because the clinical trial period will only capture part of the treatment's effect in terms of QALYs gained, since benefits achieved during the trial will have an effect beyond it, the cost-effectiveness analysis involves modeling over the longer term using complementary data. METHODS: A Markov model with states based on disability expressed by EDSS scores was used. Transition probabilities were calculated directly from clinical trial data for the first 3 years and then extrapolated to 10 years. Mean costs and utilities for each Markov state were calculated from a population-based cross-sectional study in Sweden. RESULTS: The incremental cost per QALY is SEK 342,700 (US $39,250; US $1 = SEK 8.73, March 10, 2000) when all costs (direct, informal care, and indirect) are included (discounted 3%). When indirect costs are excluded, the cost per QALY is SEK 542,000 ($62,100). CONCLUSIONS: Cost-effectiveness analysis in SPMS requires that the effect of treatment beyond clinical trials be included. Also, analysis should be done from a societal perspective, since many of the costs occur outside the healthcare system. The cost-utility ratios estimated in this analysis are at or below the mean threshold value indicated in a recent survey of health economists ($60,000). PMID- 11028133 TI - Health technology diffusion rates. Statins, coronary stents, and MRI in England. AB - OBJECTIVE: To analyze the rates and influences on the adoption of three selected health technologies: statins, coronary stents, and magnetic resonance imaging (MRI). METHODS: A retrospective diffusion study using primary care prescribing data and questionnaire responses from acute hospital trusts in the West Midlands region (population 5.3 million or 10% of England). RESULTS: The selected technologies had markedly different diffusion curves. Statins diffused rapidly soon after launch. Coronary stents were initially used 6 years after first availability, but within 2 years all responding hospitals reported using them. MRI scanners were initially purchased 6 years after first availability with a subsequently slow rate of diffusion, and are still absent from some hospitals. Influences on the adoption of each technology were different. Commercial marketing was reported as a major influence on the diffusion of statins but not at all on MRIs. Cost impact was a major negative influence on the diffusion of MRI scanners and statins, whereas enthusiastic individuals were key to the diffusion of stents. CONCLUSIONS: Influences on adoption and consequent diffusion rates are very different for different health technologies. It is not at all clear that such diffusion patterns relate well to an optimum timing rate. This has important implications for technology gatekeepers in health care. PMID- 11028134 TI - Estimating the impact of a diffuse technology on the running costs of a hospital. A case study of a picture archiving and communication system. AB - OBJECTIVES: This paper considers the methodological problems that arise in conducting cost analyses in economic evaluations where only observational (rather than experimental) data are available and where the technology being evaluated is diffuse, such that the unit of analysis has to be the institution rather than the patient. METHODS: A case study is reported that concerns the application of computer technology in radiology: picture archiving and communication systems (PACS). A range of different approaches were used to estimate changes in running costs, including time series analyses of routine data and direct observation of resource use. RESULTS: The analysis illustrates some of the difficulties involved in costing the introduction of a diffuse technology. Nevertheless, it provides a firm indication that, overall, the introduction of PACS was found to be associated with a significant increase in hospital costs, suggesting that the initial expectations of financial savings were unduly optimistic. CONCLUSIONS: The research demonstrates that, using multiple methods, it is possible to estimate cost changes within a single hospital. In addition, the paper discusses the nature of the uncertainties in such analyses and possible ways of representing such uncertainty in terms of confidence intervals. PMID- 11028135 TI - A decision model and cost-effectiveness analysis of colorectal cancer screening and surveillance guidelines for average-risk adults. AB - OBJECTIVES: Guidelines for colorectal cancer screening and surveillance in people at average risk and at increased risk have recently been published by the American Gastroenterological Association. The guidelines for the population at average risk were evaluated using cost-effectiveness analyses. METHODS: Since colorectal cancers primarily arise from precancerous adenomas, a state transition model of disease progression from adenomatous polyps was developed. Rather than assuming that polyps turn to cancer after a fixed interval (dwell time), such transitions were modeled to occur as an exponential function of the age of the polyps. Screening strategies included periodic fecal occult blood test, flexible sigmoidoscopy, double-contrast barium enema, and colonoscopy. Screening costs in 1994 dollars were estimated using Medicare and private claims data, and clinical parameters were based upon published studies. RESULTS: Cost per life-year saved was $12,636 for flexible sigmoidoscopy every 5 years and $14,394 for annual fecal occult blood testing. The assumption made for polyp dwell time critically affected the attractiveness of alternative screening strategies. CONCLUSIONS: Sigmoidoscopy every 5 years and annual fecal blood testing were the two most cost effective strategies, but with low compliance, occult blood testing was less cost effective. Lowering colonoscopy costs greatly improved the cost-effectiveness of colonoscopy every 10 years. PMID- 11028136 TI - Cost-benefit analysis of mammography screening in Denmark based on discrete ranking data. AB - OBJECTIVE: Economic evaluations such as cost-effectiveness and cost-utility analyses generally fail to incorporate elements of intangible costs and benefits, such as anxiety and discomfort associated with the screening test and diagnostic test, as well as the magnitude of utility associated with a reduction in the risk of dying from cancer. This paper seeks to include all costs and effects incurred by introducing mammography screening through the application of discrete ranking modeling. METHODS: In the present analysis, 207 women were interviewed and asked to rank, according to priority, a number of alternative breast cancer screening setups. The alternative programs varied with respect to number of tests performed, risk reduction obtained, probability of a false-positive outcome, and extent of copayment. Using discrete ranking modeling, the stated preferences were analyzed and the relative weighting of the program attributes identified. For a range of hypothetical breast cancer programs, relative utilities and corresponding willingness-to-pay estimates were derived. RESULTS: A comparison of cost and willingness to pay for each of the programs suggested that net benefits are maximized when screening person aged 50-74 years biennially. More intensive screening produces lower or similar levels of utility at a higher cost. CONCLUSION: Discrete ranking modeling can aid decision making by identifying inferior healthcare programs, i.e., programs that are more costly but less beneficial. PMID- 11028137 TI - Estimating the microvascular benefits of screening for type 2 diabetes mellitus. AB - OBJECTIVE: To define the relative benefits of screening for diabetes and improved treatment programs and ways of improving the efficiency of screening for a population-based cohort derived from the Third National Health and Nutrition Examination Survey (NHANES III). METHODS: A Markov decision model is used to estimate microvascular benefits of glucose control for four different screening and treatment scenarios, including either universal screening or improved glucose control of known diabetic subjects, neither, or both. RESULTS: A population cohort of subjects with recent onset of diabetes (< 5 years) was derived from NHANES III (of whom close to half were unaware that they had diabetes). In this population-based cohort, the total benefit achievable by universal screening and improved treatment (limiting HbA1c to less than 9%) is a reduction of about 30,000 cases of blindness over the lifetime of the cohort. Screening alone results in 7% of this benefit, and improved treatment alone provides 65%. Screening a targeted group of patients with three or more risk factors for developing diabetes would reduce the number of required fasting glucose measurements needed by 82% and provide 50% of the total benefit of screening the entire population with a fasting glucose measurement. CONCLUSIONS: Morbidity from type 2 diabetes can be most effectively reduced by developing ways to modestly improve the glycemic control of known diabetic subjects, particularly those with high A1c's and early onset of disease. Targeting can significantly reduce the number of persons who need to be screened with a fasting blood test while preserving a large component of the benefit of screening. PMID- 11028138 TI - Sensitivity analysis of the diagnostic value of endoscopies in cross-sectional studies in the absence of a gold standard. AB - OBJECTIVES: The evaluation of the diagnostic value of endoscopic procedures usually lacks a gold standard when performed in cross-sectional studies. The objective is to demonstrate an easily applicable method to assess the possible range of sensitivity, specificity, and predictive values of endoscopic procedures in the absence of a gold standard method. METHODS: Data from a study of 328 endoscopies comparing two different methods to diagnose superficial bladder cancer were used as a numerical example. Both endoscopic procedures were performed in the same patients in one session. Under the assumption of a systematic misclassification process, a model to correct sensitivity estimates is developed. RESULTS: The lowest possible sensitivity estimate for a new fluorescence endoscopy technique (FE) was 78%, the maximum 97.5%. Depending on realistic assumptions made upon the misclassification, a reasonable estimate for sensitivity was 93.4% (95% confidence interval [CI]: 90%-97.3%) for the FE technique. The sensitivity of the traditional white-light endoscopy method ranged from 47.2% to 53%, with a reasonable estimate of 46.7% (95% CI: 39.4%-54.3%). CONCLUSIONS: This method to determine the theoretically possible range of sensitivity estimates in endoscopic procedures is helpful in cross-sectional studies with a missing gold standard method. It is easily applicable for a variety of endoscopic procedures, including upper and lower gastro-intestinal tract, urogenital tract, or diagnostic laparoscopic surgery. PMID- 11028139 TI - Home rehabilitation after stroke. Reviewing the scientific evidence on effects and costs. AB - OBJECTIVES: The question addressed here is whether home rehabilitation after stroke is better and/or less expensive than the more conventional alternatives, i.e., rehabilitation during inpatient care, day care, and outpatient visits- alone or in combinations appropriate to disease stage and patient needs. Home rehabilitation is managed by teams of professionals who train patients at home. METHODS: The scientific literature was systematically searched for controlled studies comparing outcomes and costs of home rehabilitation with the more conventional strategies. RESULTS: The abstracts of 204 papers were evaluated, from which 89 were selected for greater scrutiny. From the 89 studies, we found 7 controlled studies involving 1,487 patients (6 of the 7 were randomized, 4 of the 6 assessed costs). No statistically significant differences, or tendencies toward differences, were revealed as regards the outcome of home rehabilitation versus hospital-based alternatives. Thus, home rehabilitation was neither better nor worse at improving patients' ability to manage on their own or resume social activities. Depression and reduced quality of life were common in all groups of patients and caregivers, irrespective of the rehabilitation strategy. In the four randomized studies that reported on costs, home rehabilitation was found to be less expensive than regular day care, but not less expensive than conventional strategies even though hospital stay was reduced. CONCLUSION: The outcomes and costs of home rehabilitation after stroke seem to be comparable to alternative treatment strategies. PMID- 11028140 TI - Motivated patients are more cost-effectively rehabilitated. A two-year prospective controlled study of patients with prolonged musculoskeletal disorders diagnosed in primary care. AB - OBJECTIVES: To compare the cost-effectiveness of a multidisciplinary rehabilitation program with traditional treatment within primary care in terms of the health-related quality of life (HRQOL) in patients with prolonged musculoskeletal disorders (MSD) on the one hand and the costs to society on the other. Predictors of total costs, such as motivation, socio-economic level, age, pain, and working environment, were investigated. METHODS: A prospective, matched, controlled 2-year follow-up study was designed. The main outcome measures were HRQOL using the Nottingham Health Profile (NHP) and patient specific total costs due to society. The cost-effectiveness was expressed as a quotient of the total costs to society/NHP global score difference value. RESULTS: Patients with prolonged MSD generate substantial total costs to society, chiefly in the area of indirect costs. Multidisciplinary rehabilitation improved HRQOL more cost-effectively. Motivation was revealed as a predictor of total costs. The relationship in savings in terms of indirect costs between the highly motivated and the less-motivated patients was calculated at 4:1. CONCLUSIONS: The large group of patients with prolonged MSD generate substantial total costs, and even small reductions in direct and indirect costs could be of importance to society. The multidisciplinary rehabilitation program applied here was more cost effective as compared with conventional treatment within primary care when it came to improving the patients' perceived HRQOL. Motivation could be a predictor of total costs, which has to be addressed in the process of identifying the patient as a partner in the rehabilitation process. PMID- 11028141 TI - The effect of cochlear implant use in postlingually deaf adults. AB - OBJECTIVE: To assess the effect of the use of cochlear implants (CI) on the health status of postlingually deaf adults. METHODS: Participants comprised 45 postlingually deaf adult multichannel CI users and 46 deaf candidates on the waiting list for a CI. The latter group acted as control subjects to corroborate the validity of retrospective completion of the questionnaires by the CI recipients. Three HRQOL instruments were used: a) a specially developed CI questionnaire (NCIQ); b) a generic HRQOL questionnaire (SF-36); and c) a health state classification system (HUI-2) suited to estimate single preference scores. RESULTS: Retrospectively estimated pre-implant scores in the CI user group corresponded very well with the scores in the control group. Postimplant scores in the CI users were substantially higher in all six domains (p < .001) of the NCIQ than the scores in the control group. Effects due to a CI were also observed with the SF-36 in five of the seven domains (p < .01). Statistically significant differences between the two groups (p = .001) were observed in two of the six domains of the HUI-2. CONCLUSIONS: All three questionnaires detected improvements in HRQOL due to CI use. To make a detailed assessment of the effect of a CI on functional outcomes and well-being, a special purpose HRQOL instrument is far more adequate than a general HRQOL instrument. This study also showed that a CI affects several other health domains besides auditory performance. The effect of CI use on general functioning and well-being proved to be considerable. PMID- 11028142 TI - Allogeneic stem cell transplantation. An economic comparison of bone marrow, peripheral blood, and cord blood technologies. AB - OBJECTIVES: To compare outcome attributes of three available technologies for stem cell transplantation--bone marrow transplantation (BM), peripheral blood cell transplantation (PB), and cord blood transplantation (CB). To compare the cost per recipient for the three technologies, incorporating all donor and antigen matching functions, as well as the transplantation procedures. METHODS: Review of relevant literature using the CancerLit and HealthSTAR databases (plus earlier searches of MEDLINE and Embase) to summarize outcome differences. Estimations of cost per transplant using models drawing on Canadian experience. RESULTS: The literature review indicated that PB may have a modest advantage over BM in terms of outcomes. The literature indicated that CB has longer engraftment times (although this may be due to inadequate controls). Our modeling exercise indicated that costs for PB and BM are similar. CB has a higher cost per recipient, due in large part to the higher testing costs for every donated unit. CONCLUSIONS: Current literature is inadequate to provide an outcomes comparison of CB, PB, and BM. Viewed from a system-wide perspective, CB is a higher cost technique. However, PB and CB are emerging as technologies, and so our results should be viewed alongside the notion that there may be a steep learning curve in these technologies. PMID- 11028143 TI - Unjustified variations in patient management for acute myocardial infarction in the Rhone-Alpes region of France. PRIMA Group. AB - OBJECTIVES: To examine determinants of use of cardiac procedures after acute myocardial infarction and identify variation factors. METHODS: Observational prospective cohort study of 2,519 patients in 48 centers with a two-level logistic-regression analysis. RESULTS: Elderly patients were less likely to undergo pre- and inhospital thrombolysis (odds ratios, 0.71 and 0.64; 95% CI, 0.62-0.81 and 0.58-0.69, respectively). The elderly, females, and patients with heart failure on admission were less likely to undergo noninvasive tests (0.74, 0.62, and 0.51; 95% CI, 0.67-0.81, 0.46-0.83, and 0.38-0.68, respectively) and coronary angiography (0.38, 0.53, and 0.67; 95% CI, 0.34-0.42, 0.38-0.74, and 0.52-0.86, respectively) but not revascularization. Hospital factors were more difficult to interpret. CONCLUSIONS: Elderly, women, and heart failure patients underwent fewer cardiac procedures than lower-risk patients. Physicians should change their attitude toward these groups and use advanced procedures, bearing in mind the patients' needs rather than good procedural outcomes. PMID- 11028144 TI - Health technology assessment in Canada. A decade in review. AB - OBJECTIVES: Since 1988, four government-funded health technology assessment (HTA) agencies have been established in Canada. This paper is a descriptive review of reports issued by these organizations during the period from 1988 to 1998. METHODS: Publications from the national and three provincial HTA agencies in Canada were obtained and reviewed. Only the 117 assessment reports that were reported to have undergone external review were included in this analysis. Each report was classified on a standard abstraction form according to criteria such as technology type(s), assessment focus, whether a specific policy question was clearly stated and relevant decision maker(s) identified, description of search strategy and selection criteria, sources of data and assessment methods used, whether recommendations or conclusions were made, and duplication or overlap of reports. The trends in these qualities over the 10-year period were also examined. RESULTS: Therapeutic technologies have received the most attention from all four agencies, although the focus on devices, drugs, and procedures varied between agencies. The policy question under investigation was clearly identified in 82% of reports, and 71% clearly identified the decision maker toward whom the assessment was targeted. Efficacy or effectiveness was examined in 81% of reports, and costs were assessed in 65% of studies. These were the two most frequently examined aspects. Almost all assessments were descriptive literature reviews; 9% included meta-analyses and 32% had cost analyses or economic evaluations. Forty-four percent of reports had a clear description of the literature search strategy, and selection criteria were clearly specified in 38% of studies, but there was considerable variation among agencies in the level of description of these methods. Conclusions were clearly stated in 83% of the assessments' conclusions, and 13% had recommendations. When analyzed longitudinally, it is apparent that the quality of reports has improved markedly during the past decade. This was determined by examining the clarity of specifying the policy question(s) under investigation, the identification of the target audience of decision makers for the information, and by evaluating the thoroughness of the description of the methods used in the assessment. CONCLUSIONS: Canadian government agencies have contributed a considerable quantity of health technology assessments. There has been very little duplication of technologies evaluated, and the quality of the assessment reports has markedly improved during the past decade. PMID- 11028145 TI - Health technology assessment at the end of life. A realistic view of death and palliative care. PMID- 11028146 TI - A study of the references used in Cochrane protocols and reviews. Three bibles, three dictionaries, and nearly 25,000 other things. AB - OBJECTIVES: To describe the types of report that are cited by protocols and reviews included in the Cochrane Database of Systematic Reviews. METHODS: The citation for each reference included in the Cochrane protocols and reviews published in the Cochrane Database of Systematic Reviews in issue 1, 1999 of The Cochrane Library were categorized by reference type (e.g., journal article, conference proceeding, book chapter, personal communication, Cochrane review). RESULTS: Of a total of 24,913 citations, 21,694 (87.1%) were references to journal articles. There was a significant difference between the proportion of references to studies that were journal articles (12,348 of 13,472; 91.7%) and the proportion of other references in this category (9,346 of 11,441; 81.7%). CONCLUSION: The great majority of studies included in Cochrane reviews at the beginning of 1999 had been published as journal articles. PMID- 11028147 TI - Cardiac pacemakers. When clinical evaluation lags behind technological progress. AB - OBJECTIVE: The rise in the number of implantations of cardiac pacemakers is of some concern to decision makers in the health sector. We assessed the intrinsic and relative clinical efficacy of cardiac pacemakers in current clinical indications to find out whether scientific or clinical arguments might justify differences in market prices. METHODS: We retrieved papers on cardiac pacing (January 1993-April 1998) from five databases (MEDLINE, HealthSTAR, EMBASE, Cochrane Library, and PASCAL). The citations in these papers were used to seek further articles. We selected the articles that met the criteria of evidence based medicine (EBM) (randomized and nonrandomized controlled trials) and classified them according to clinical indication and type of evaluation (either of the intrinsic efficacy of a pacemaker versus a control or of the relative efficacy of different pacing modes). RESULTS: A total of 542 references were retrieved, but under 10% met our EBM criteria. Very few were comparative studies versus controls; most were recent and tended to use endpoints other than survival. Clinical efficacy was not proven on the basis of EBM criteria, even in common indications (e.g., sick sinus syndrome). Studies comparing different pacing modes were rarely randomized and did not provide consistent evidence for the superiority of any pacing mode in a given indication. CONCLUSIONS: Knowledge of the natural history of the diseases for which cardiac pacing is indicated is scarce. There is an approximately 20-year gap between technological progress and clinical evaluation that cannot be easily bridged because of methodologic difficulties and ethical issues. Current guidelines on pacemaker use either rely on expert opinion or highlight present inadequacies and make recommendations for future work. Available clinical efficacy data do not justify the wide differences in the price of cardiac pacemakers. PMID- 11028148 TI - Mini-summaries from the U.K. HTA programme. PMID- 11028149 TI - Reports from the Swedish Council on Technology Assessment in Health Care. Back pain. PMID- 11028150 TI - The literature search for Bloom et al.'s review. PMID- 11028151 TI - The value of free and total prostate specific antigen in identifying patients for prostatic biopsy and its relationship to Gleason score and number of positive cores. AB - INTRODUCTION AND OBJECTIVES: The percentage of free over total prostate specific antigen (%F/T PSA) seems to enhance the predictive value of PSA in diagnosing prostate cancer. We evaluated the value of F/T PSA in 50 consecutive patients who underwent prostate needle biopsy, its relationship to the Gleason score and number of positive cores. MATERIAL AND METHODS: 50 patients underwent prostate needle biopsy for abnormal PSA and/or abnormal digital rectal examination (DRE). There were 8 patients with PSA equal or less than 4 ng/ml, 6 with F/T PSA < 20%, group I (GI). 27 patients with PSA between 4.1 ng/ml and 10.0 ng/ml, 20 with F/T PSA < 20%, group II (GII) and 15 patients with PSA > 10.1 ng/ml (13 with F/T PSA < 20%), group III (GIII). At least six needle biopsies were obtained guided by transrectal ultrasound selectively or randomly. Pathological evaluation included Gleason grade and number of cores involved. RESULTS: 21/50 patients (42%) had positive biopsies, 3/8 in GI, 8/27 in GII (6 had negative DRE) and 10/15 in GIII (9 had positive DRE). 19/21 patients with positive biopsies had F/T PSA < 20%. The sensitivity, specificity and positive predictive value of PSA between 4-10 ng/ml and F/T PSA < 20% was 87.5%, 31% and 35% respectively. Stratifying patients with positive biopsies to F/T PSA < 10%, F/T PSA > 10% and the three PSA groups, there was no relationship to either Gleason score or number of positive cores. CONCLUSION: With a cutoff of 20%, F/T PSA seems to be an important parameter in selecting patients with abnormal PSA for biopsy. It will be helpful mostly with PSA 4-10 ng/ml. No relationship was observed between the level of F/T PSA, grade or number of positive cores. PMID- 11028152 TI - In-hospital mortality after acute myocardial infarction in Lebanon: incidence, associations, and influence of newer treatment regimens. AB - PURPOSE: To study the incidence of in-hospital mortality following acute myocardial infarction in Lebanon, and its relationship to demographic, clinical variables, and therapeutic strategies. PATIENTS & METHODS: Consecutive admissions due to myocardial infarction to 18 medical centers in various regions of Lebanon were entered into the Lebanese Myocardial Infarction Study, conducted between January and July 1996. Information was obtained on age, gender, time of onset of symptoms, delay to hospital arrival, mode of transport, and coronary risk factor analysis. The patients were followed up in hospital for analysis of modes of therapy, complications and mortality. RESULTS: There were 44 in-hospital deaths among the 433 admissions (10.2%), which is a rate lower than those previously reported from Lebanon. Less than half the patients presented within 6 hours of onset of symptoms and only 28% received thrombolytic therapy. The mortality rate was higher in older age groups, those presenting with cardiogenic shock or pulmonary edema, those with a history of angina, infarction or heart failure, and those who developed recurrent ischemia or infarction during their hospital stay. Furthermore, occurrence of ventricular arrhythmias, mechanical complications, congestive heart failure and left bundle branch block was associated with a higher mortality rate. Treatment with angiotensin converting enzyme inhibitors, beta-blockers, aspirin, heparin, nitrates and thrombolytics significantly reduced mortality rates. CONCLUSIONS: The results reveal improvement in the survival of patients after acute myocardial infarction to values similar to those reported from Western countries. Further effort should be expended to enhance early arrival to the hospital, increased thrombolytic therapy and to implement treatment strategies supported by large clinical trials such as use of aspirin, ACE inhibitors and beta-blockers. PMID- 11028153 TI - [Stereotaxic excisional biopsy of non-palpable breast lesions by the ABBI(Advanced Breast Biopsy Instrumentation) technique. Advantages. Disadvantages. Indications. Apropos of 67 cases]. AB - OBJECTIVES: Introduce briefly: the classification of the American College of Radiology for non palpable breast lesions; and the different techniques used for stereotactic breast biopsies. Evaluate the efficiency of the ABBI technique for non palpable breast lesions and its role in breast biopsies, on the base of 67 cases. SUBJECTS AND METHOD: Sixty-seven stereotactic excisional biopsies on dedicated prone tables were performed using the ABBI technique for 21 probably benign and 46 indeterminate non palpable breast lesions seen on mammograms. Twenty-five cases were densities, architectural distortions or radial scars, and 42 were microcalcifications. RESULTS: In our hands, the technique is 100% specific and sensitive. This high percentage of success is related to a team work between radiologists, surgeons or gynecologists. The specimen represent the whole lesion. Ductal carcinoma in situ (DCIS) and atypic ductal hyperplasia (ADH) are diagnosed with accuracy. Twelve carcinomas are found, three of them DCIS. ADH was encountered in two cases. Complications are minor, hematomas of small and moderate volume were encountered in eight cases (12%). CONCLUSION: ABBI technique replaces surgical biopsies after wire hook localisation for solitary lesion smaller than two cm in diameter; and stereotactic Tru-cut biopsies, even with vacuum aspiration, for small cluster of microcalcifications, architectural distortion and radial scar, where the percentage of failure is higher. For malignant lesions, surgery must follow, even when the margins are free. PMID- 11028155 TI - Hypertension in the elderly. Part I. PMID- 11028154 TI - Pattern of proven bacterial sepsis in a neonatal intensive care unit in Riyadh Saudi Arabia: a 2-year analysis. AB - Bacterial infections are an important cause of neonatal mortality and morbidity. The major pathogens for neonatal sepsis in the neonatal intensive care unit (NICU) vary with geographical area and time. It is therefore important to frequently audit neonatal sepsis in individual NICU, to aid in provision of adequate and appropriate preventive and therapeutic measures. We retrospectively reviewed the medical records of all infants who had positive blood cultures during a 2-year period in the NICU at a university hospital in Riyadh, Saudi Arabia. Overall the incidence of proven-bacterial-sepsis (PBS) was 10.2% of NICU admissions. The incidence of PBS in low-birth-weight (LBW), very low-birth-weight (VLBW), and extremely low-birth-weight (ELBW) infants were 19%, 41%, and 49% respectively. Multiple episodes of bacterial sepsis occurred in 21% of all infants infected. Coagulase negative Staphylococcus (CONS) (50%) was the most common infecting organism causing late onset sepsis (LOS) and Escherichia coli (29%) the most common causing early onset sepsis (EOS). Gram negative bacteria (GNB) were the infecting organisms in 50% of the EOS episodes and 29% of LOS episodes. Only 11% (14) of the PBS were EOS. Only 10 (10.4%) infants had bacterial meningitis. The overall PBS related mortality was 9%, representing 22% of all neonatal deaths. PMID- 11028156 TI - The promise of gene therapy as a therapeutic modality in heart failure. PMID- 11028157 TI - Snoring and obstructive sleep apnea syndrome a multidisciplinary approach. The role of the dentist. PMID- 11028158 TI - [Endoscopy: first images]. PMID- 11028159 TI - [Familial lymphohistiocytosis. Evolution of management apropos of 3 cases]. AB - Familial lymphohistiocytosis is a rare rapidly lethal genetic disease. It is characterized by an uncontrolled activation of T lymphocytes and macrophages, with multiple organ infiltration, beginning with fever and unexplained coagulopathy. Recently, one of the genes implicated in 50% of families at risk was identified (locus FHL1, chromosome 10, region q21-22). Based on data suggesting an essential role of T lymphocytes in the genesis of familial lymphohistiocytosis, the treatment has recently evolved from a chemotherapy including Etoposide (VP16) and corticosteroids, sometimes efficient but toxic, to an almost always efficient and slightly toxic immunosuppressive treatment. These two treatments achieved a remission somewhat lasting with no definite cure. In fact, all patients relapsed in the central nervous system and died. Bone marrow transplantation (BMT) is the only curative treatment. However only 20% of patients benefit from an HLA identical BMT. Recent improvements in HLA non identical BMT offer an acceptable alternative to the other 80% of patients. In this review, we present three cases illustrating the evolution and optimization in the management of infants with familial lymphohistiocytosis. PMID- 11028160 TI - [Hemostatic embolization of hepatocellular carcinoma complicated by hemoperitoneum. Apropos of 2 cases and review of the literature]. AB - Rupture of hepatocellular carcinoma is a severe complication that occurs in about 10% of patients. It may occur as a terminal event in patients with advanced disease or it may be the first presentation in a healthy individual. Various treatment options have been proposed, which include conservative treatment, transarterial embolization and operative hemostasis or liver resection. We report intraperitoneal hemorrhage and hypovolemia in two patients with spontaneous rupture of an hepatocellular carcinoma treated successfully by transarterial hepatic embolization. On follow-up, these patients died 7 and 8 months after this treatment respectively. PMID- 11028161 TI - Neurological complications associated with Mycoplasma pneumoniae infection. A case report. AB - Mycoplasma pneumoniae (MP) is a frequent cause of respiratory tract infection. Extra-pulmonary manifestations may be neurological with variable expression, encephalitis being the most frequent. Meningitis, myelitis or polyradiculoneuritis are also reported. The pathophysiology of neurological manifestations remains poorly understood. Although not isolated from cerebral tissue, MP was reportedly detected in cerebro-spinal fluid (CSF) on several occasions. We report the case of a five-year-old girl who presented with fever, then pneumonia, and later developed spastic quadriparesis. MP was identified as the causative agent, evidenced by the presence of specific antibodies in both serum and CSF. The identification of a central nervous syndrome associated with confirmed MP infection extends the list of similar cases documented in individuals younger than 17 years of age. PMID- 11028162 TI - [Medical confidentiality in Lebanon (II)]. PMID- 11028163 TI - [Homocysteine and its metabolites in chronic renal insufficiency and the effect of a vitamin replacement]. AB - BACKGROUND: Hyperhomocysteinemia has been increasingly recognized as an important risk factor for elevated atherosclerotic vascular disease in chronic renal failure. We measured in patients with chronic renal failure homocysteine and metabolites of its 2 metabolic pathways, transulfuration (cystathionine, cysteine) and remethylation (methionine, methylmalonic acid, 2-methylcitric acid). PATIENTS AND METHODS: Eleven patients on conservative treatment (creatinine clearance 10 to 30 ml/min) and 50 chronic uremic subjects on regular hemodialysis were included in the study. Twenty-two of the dialysis patients received daily oral multivitamin supplementation containing 10 mg vitamin B6, 6 micrograms vitamin B12, and 1 mg folic acid during the last year before the study started. RESULTS: In the hemodialysis group homocysteine levels were higher compared with the patients on conservative treatment. Hemodialysis patients with additional vitamin supplementation showed significantly lower homocysteine levels than those without. The pattern of metabolites was different to these results: all metabolites were higher in hemodialysis patients, too (significant for cysteine and methionine), but vitamin supplementation failed to lower all metabolites. CONCLUSION: Analysis of metabolites additional to homocysteine levels may help to understand different results in evaluation of atherosclerotic risk of hyperhomocysteinemia in chronic renal failure. PMID- 11028164 TI - [Opportunistic infections and HIV-associated malignancies. An evaluation of 58 autopsy cases within 10 years]. AB - PATIENTS AND METHODS: Of all patients who died of HIV infection within 10 years (1. 1. 1988 to 31. 12. 1997) at the Klinikum Nurnberg 58 autopsy cases were reviewed at the Institute of Pathology of the above mentioned hospital. RESULTS: The male:female ratio was 2.1:1, the mean age being 40.5 years. Most of the patients showed an advanced stage of lymphadenopathy at the moment of death. Non Hodgkin's lymphoma and Kaposi's sarcoma, both HIV-related malignant diseases, were diagnosed in 6/58 cases, 10.3% each. HIV-related myelodysplastic changes existed in 28/58 patients (48.3%). Twelve patients showed an HIV-associated encephalopathy (20.7%). Opportunistic infections (pneumocystis carinii, cytomegaly, toxoplasmosis, atypical mycobacteriosis) were found in 28/58 patients (48.3%). A mycosis was diagnosed in 9/58 cases (15.5%). Tuberculosis was identified in 4/58 patients (6.9%). Cirrhosis of the liver was ascertained in 8/58 patients (13.8%). 24/58 patients (41.4%) died of respiratory disorder. CONCLUSION: In the acquired immunodeficiency syndrome numerous morphological findings will be helpful in diagnosis and therapy. PMID- 11028165 TI - [Cardiac complications in diabetes mellitus]. AB - Diabetes mellitus as a disease of epidemiological impact leads to diabetic cardiopathy by modulation of myocardial, vascular and metabolic components. This includes the development of a coronary microangiopathy and a decrease of diastolic and systolic function of the left ventricle as well as the development of an autonomic diabetic neuropathy. Patients with diabetes show an increased mortality concerning cardiovascular events. They more often suffer from myocardial infarction as non-diabetics mostly with a more serious course. Moreover, the post-infarction course is affected with a worse prognosis as in non diabetics. For diagnosis of cardial involvement in diabetes electrocardiographic and echocardiographic procedures are of use. Special tests of the autonomic function complete the diagnostic ensemble. An early therapy with ACE-inhibitors and beta blocking agents as well as a strong diabetes therapy, in particular with insulin, can influence the mortality favorably. Moreover, the diagnosis and therapy of additional cardiovascular risk factors (arterial hypertension, dyslipidemia) are very important, because these are correlated with a for diabetic patients markedly increased risk of mortality. The clinical relevance of the term diabetic cardiopathy is justified by the 6 factors: macroangiopathy, microangiopathy, disturbances of the myocardial metabolism, myocardial fibrosis, autonomic diabetic neuropathy and disturbances of the coagulability. Diagnostic and therapeutic goals are discussed. PMID- 11028166 TI - [Antimicrobial peptides and peptide antibiotics]. AB - Antimicrobial peptides are naturally occurring antibiotics. As part of the innate immune system of vertebrates they have direct antimicrobial function. Further, they can act as mediators of inflammation. Their antimicrobial spectrum covers gram-positive and -negative bacteria as well as fungi and certain viruses. Based on their structure, antimicrobial peptides can be divided into several families. Peptides of the defensin, cathelicidin, and histatin families have been isolated from humans, where they have been found in defense cells, such as macrophages or neutrophils, as well as in epithelial cells. Decreased production of antimicrobial peptides is associated with immune deficiencies. Further, lung disease in cystic fibrosis may be linked to the dysfunction of antimicrobial peptides. Based on naturally occurring antimicrobial peptides, derivates of these molecules were developed as innovative antibiotic drugs. The present review focuses on the biology of antimicrobial peptides as well as their potential as drugs. PMID- 11028167 TI - [Update endocrinology I (Neuroendocrinology)]. AB - BACKGROUND: Presentation of current data and knowledge in the field of experimental and clinical neuroendocrinology. AIM: Overview of current data on the pathogenesis and therapeutic strategies of pituitary disorders. 1. Hypothalamic and pituitary regulation: characterization of influence of hormone secretion and cell proliferation of anterior pituitary cells by an intrapituitary network of cytokines and growth factors. Special importance on the role of the interleukin-6-family. Characterization of the role of gaseous transmitters in neuroendocrinology. 2. Molecular pathogenesis of pituitary adenomas: presentation of the current knowledge status with regard to familial syndromes. 3. Therapy of pituitary adenomas: presentation of new developments of therapeutic aspects, in particular medical treatment, surgery and radiation. Outlook on the future role of gene therapy. 4. Growth hormone replacement in adults: short overview of diagnosis and therapy of growth hormone deficiency in the adult. PMID- 11028168 TI - [Therapy-resistant ventricular tachycardia in a woman with primary cardiac non Hodgkin lymphoma]. AB - HISTORY: A 65-year-old woman had suffered from relapsing ventricular tachycardias (VT) since 1996. FINDINGS: Physical examination was normal. An arrhythmogenic substrate was found in the right ventricular outflow tract by electrophysiological examination. Nuclear magnetic resonance imaging (MRI) showed an infiltration of the right heart. Myocardial biopsy revealed a high-grade centroblastic non Hodgkin lymphoma. The patient was now transferred to our hospital for further treatment. Lactate dehydrogenase was elevated (2,030 U/l). Echocardiography showed a thickened and more reflecting right ventricular myocardium. Bone marrow aspiration and MRI/computed tomography of abdomen and thorax excluded a generalized stage. Ventricular tachycardias were caused by a primary cardiac lymphoma. TREATMENT AND COURSE: Combined radio-chemotherapy succeeded in complete remission. High-frequency ablation and amiodarone failed. Although MRI showed no more vital lymphoma after the combined radio-chemotherapy the patient suffered from spontaneous and symptomatic relapses of VT. Therefore this patient with primary cardiac lymphoma was the first in literature to get a defibrillator (ICD). The incidence of VT decreased and up to now the patient showed no relapse of the non Hodgkin lymphoma (follow-up 23 months). PMID- 11028169 TI - [Recurrent ventricular fibrillation after defibrillation in a patient with pacemaker]. AB - HISTORY: A 62-year-old patient was admitted for an operation of a hernia of the right groin in the surgical department. He had a dual chamber pacemaker because of a binodal disease. Pre-operatively the patient was reanimated because of ventricular fibrillation in dilated cardiomyopathy and was defibrillated 12 times, unexpectedly often. DIAGNOSIS: The reason of the repeated ventricular fibrillation was a spike-on-T-phenomenon due to loss of the sensing function of the pacemaker (entrance bloc). TREATMENT: Therefore, the pacemaker was explanted, and the ventricular fibrillation came to an end. The patient recovered. CONCLUSIONS: The repeated defibrillations with the electrodes over the basis and the apex of the heart damaged the pacemaker. The following fibrillation episodes were founded on a spike-on-T-phenomenon. In order to prevent such damage of pacemakers, pacemaker patients should be defibrillated and cardioverted in anterior-posterior position of the electrodes. PMID- 11028170 TI - [Castleman disease]. AB - BACKGROUND: The term "Castleman's disease" (angiofollicular lymph node hyperplasia) comprises a heterogeneous clinicopathologic entity among lymphoproliferative disorders. CASE REPORT: We present a rare case with retroperitoneal manifestations, and discuss several aspects of diagnosis, differential diagnosis, the clinical course and therapy of the disease. Finally, we summarize the actual knowledge about the pathogenesis of Castleman's disease. PMID- 11028171 TI - [Subclavian steal syndrome]. PMID- 11028172 TI - [Quality assurance in medicine--who does it serve?]. PMID- 11028173 TI - [Life expectancy in Down syndrome infants and children with congenital heart defects, 1974-1997]. AB - Authors analysed 359 cases with Down's syndrome and congenital heart defects registered between 1974-1997 in Hungary. The total death rate was 19.9% (70 cases). Mortality in the operated group (85 cases) was 10.5% (9 patients), in the non-operated group (274 cases) 22.2% (61 patients). The death rate was lower in the group with early primary reconstruction (2.3%) than in the group with palliation + reconstruction (15.3%), or in the group with only palliative procedure (20%). These results indicate that the life expectancy of infants and children with Down's syndrome and congenital heart disease after early primary reconstructive procedure is the same as in Down syndrome patients without cardiac defects. The prognosis depends on the patient's social circumstances. The results after correct surgical procedure in patients with the same cardiac defect are similar to that of the patients with or without Down's syndrome. PMID- 11028174 TI - [Alterations of cerebral hemodynamics in pre-eclampsia and eclampsia]. AB - Clinical and pathophysiological aspects of preeclampsia and eclampsia have been found to be responsible for the majority of maternal mortality, but the exact role of them is still unknown. It is conceivable that altered cerebral hemodynamics and autoregulation is responsible for the neurological complications. The background of the cerebral processes is still not clarified. Authors summarize the changes of cerebral circulation which develop during preeclampsia and explanations for cerebral hemodynamic alterations during normal, preeclamptic and exclamptic pregnancies are discussed. They conclude that there is no accurate method for predicting eclampsia. Cerebral hemodynamical studies are only helpful in better understanding the pathophysiological background of preeclampsia-eclampsia syndrome at present. PMID- 11028175 TI - [Treatment of open wounds of the heart and the pericardium: analysis of the relation between the time factor and diagnostic procedures]. AB - Authors summarize their experiences connected with the treatment of open wounds of the heart and the pericardium during the past 35 years. Between 1962 and 1997 the authors operated on 31 cases of injuries affecting the heart. 10 wounds were situated in the isolated pericardium. They also deal with the history of treating injuries of the heart and pericardium. The value of diagnostical procedures was analysed and compared with Hungarian literature data that describes further cases. They emphasize the importance of immediate physical examination (including that of circulation, breathing, heart rate, consciousness, etc.) in cases of wounds close to the heart. Authors discuss the effects and the importance of the circumstances that directly affect the life expectancy of the patient during the treatment, such as time factor, the planning of cooperation within the operating team, practice in the field of thoracic surgery and management. They also put stress on correct thoracotomy, drainage and suction, so as to avoid postoperative pleural complications. PMID- 11028176 TI - [Fluorescence in situ hybridization in the diagnosis and follow-up of chronic myelogenous leukemia]. AB - Chronic myelogenous leukaemia is a clonal myeloproliferative stem cell disease. Its cytogenetical hallmark is the Philadelphia chromosome (Ph) or the BCR/ABL fusion gene. Their identification is important both in the diagnosis and the follow-up of the disease. In our department we have investigated the BCR/ABL gene arrangement in 21 patients with fluorescence in situ hybridization. The aim of the analysis in freshly suspected patients without any previous therapy was to confirm diagnosis and mapping the ratio of Philadelphia positive cells. In contrast to the 95-100% Ph-positivity of mononuclear cells by classical cytogenetical examinations we found BCR/ABL gene arrangement only in various but always lower proportions. Therefore the latter examination gives a better representation of residual normal hemopoesis. Out of 9 patients who had received interferon treatment for at least 6 months, 4 gave a major, 4 a minor cytogenetical answer and in 1 case there was no cytogenetical response. Seven patients reached a complete and 2 a partial hematological remission. Among 5 other patients receiving interferon treatment, in 2 cases with double Ph positivity we found a rapid progression. The data of 3 patients had to be excluded from the evaluation due to the so far short following time. PMID- 11028177 TI - [Thrombophilia in ulcerative colitis]. AB - The authors report a combined occurrence of thrombophilia and colitis ulcerosa and provide an analysis of relevant references in earlier works. It is likely intertwinning of the two disease's symptoms, the chronic cause of the condition and the underlying molecular biology variations cannot be traced back to a single cause. Further research is required to establish whether the protein-C anomaly exhibited in the presented case is general in this condition or an individual occurrence. The publication emphasises that in future cases it will be advisable to perform tests capable of proving or refuting the abnormality of protein-C. PMID- 11028178 TI - [In Process Citation] PMID- 11028179 TI - [Immunology summary or immunology in two hours]. PMID- 11028180 TI - [Respiratory infection in children]. PMID- 11028181 TI - [Effects of the environment on upper airway pathology in children]. PMID- 11028182 TI - [Recurrent ORL infections in children]. PMID- 11028183 TI - [Serological diagnosis and nasopharyngeal washings in pediatric infections]. AB - In the course of respiratory infections, the efficacy of microbiologic diagnosis has increased years after years, in term of specificity, sensitivity and rapidity. New pathogenic agents have been described such as: Legionella pneumophila, Chlamydia pneumoniae, Hantavirus. Some viruses have been well characterized as responsible for seasonal outbreaks using rapid tools for identification. Needs for efficient diagnostic tools became more obvious when specific antiviral drugs appeared on the market. So technologic developments improved the efficacy of microbiologic diagnosis and anticipate a better specificity as well as sensitivity with the help of molecular biology. Respiratory syncytial virus is one of the major infectious agents found in respiratory infections in young children and newborns. On the whole it was detected in more than one third of pediatric nasopharyngeal aspirations received in our laboratory and more than 50% during the peak of the winter epidemics. The method of direct antigen detection by immunofluorescence with the help of monoclonal antibodies allowed us to establish an incidence curve of these recurrent outbreaks, beginning in December to stop usually by the end of April. During this same period, influenza A virus, seldom influenza B virus, were detected in many nasopharyngeal specimens. Other viruses, parainfluenza 1 to 3 and Adenovirus, were irregularly detected all along the year. In the great majority of nasopharyngeal aspirations with a positive virus detection, one virus only was observed. Antigen detection methods were also developed for some bacteria such as Chlamydia pneumoniae, Legionella pneumophila. Although serology is not frequently used by pediatricians, it is still necessary for the diagnosis of Mycoplasma pneumoniae infections. A direct antigen detection test is now available, but its sensitivity needs to be evaluated. On the other hand serologic diagnosis may be extremely useful when long lasting or treatment resistant respiratory infections occur. Seroconversion or four-fold increasing titers to one pathogen may be observed when a second serum sample is tested together with the first serum of this patient. The diagnostic yield will be all the more efficient that time between both samples is long. Molecular biology techniques will significantly change the way to investigate an infection. Presently these methods are used in research laboratories, but automated technologies will facilitate routine laboratory workload. Screening methods using multiplex PCR are also promising. PMID- 11028184 TI - [Means of bacterial resistance]. AB - Fifty years ago, the introduction of penicillin, followed by many other antibacterial agents, represented an often underestimated medical revolution. Indeed, until that time, bacterial infections were the prime cause of mortality, especially in children and elderly patients. The discovery of numerous new substances and their development on an industrial scale confronted us with the illusion that bacterial infections were all but vanquished. However, the widespread and sometimes uncontrolled usage of these agents has led to the selection of bacteria resistant to practically all available antibiotics. Bacteria utilize three main resistance strategies: (i) decrease in drug accumulation, (ii) modification of target, and (iii) modification of the antibiotic. Bacteria can decrease drug accumulation either by becoming impermeable to antibiotics, or by actively excreting the drug accumulated in the cell. As an alternative, they can modify the structure of the antibiotic's molecular target--usually an essential metabolic enzyme of the bacteria--and thus escape the drug's toxic effect. Lastly, they can produce enzymes capable of modifying and directly inactivating the antibiotics. In addition, bacteria have evolved extremely efficient genetic transfer systems capable of exchanging and accumulating resistance genes. Some pathogens, such as methicillin-resistant Staphylococcus aureus and enterococci are now resistant to almost all available antibiotics. Vancomycin is the only non-experimental drug left to treat severe infections due to such organisms. However, vancomycin resistance has already appeared several years ago in enterococci, and was also recently described in staphylococci, in Japan, France and the United-States. Antibiotics are precious drugs which must be administered to patients who need them. On the other hand, the development of resistance must be kept under control by a better comprehension of its mechanisms and modes of transmission and by abiding by the fundamental rules of anti-infectious chemotherapy, i.e.: (i) choose the most efficient antibiotic according to clinical and local epidemiological data, (ii) target the bacteria according to the microbiological data at hand, and (iii) administer the antibiotic at an adequate dose which will leave the pathogen no chance to develop any resistance. PMID- 11028185 TI - [Pneumococcal antibiotic resistance]. AB - In 1875, 7 years prior to the description of the Koch bacillus, Klebs visualized the first Streptococcus pneumoniae in a pleural fluid. Since then, this organism has played a determinant role in biomedical science. From a biological point of view, it was largely implicated in the development of passive and active immunization by serotherapy and vaccination, respectively. Genetic transformation was also first observed in S. pneumoniae, leading to the discovery of DNA. From a clinical point of view, S. pneumoniae is still today a prime cause of otitis media in children and of pneumonia in all age groups, as well as a predominant cause of meningitis and bacteremia. In adults, bacteremia is still entailed with a mortality of over 25%. Although S. pneumoniae remained very sensitive to penicillin for many years, penicillin-resistance has emerged and increased dramatically over the last 15 years. During this period of time, the frequency of penicillin-resistant isolates has increased from < or = 1% to frequencies varying from 20 to 60% in geographic areas as diverse as South Africa, Spain, France, Hungary, Iceland, Alaska, and numerous regions of the United States and South America. In Switzerland, the current frequency of penicillin-resistant pneumococci ranges between 5 and > or = 10%. The increase in penicillin-resistant pneumococci correlates with the intensive use of beta-lactam antibiotics. The mechanism of resistance is not due to bacterial production of penicillinase, but to an alteration of the bacterial target of penicillin, the so-called penicillin binding proteins. Resistance is subdivided into (i) inter mediate level resistance (minimal inhibitory concentration [MIC] of penicillin of 0.1-1 mg/L) and (ii) high level resistance (MCI > or = 2 mg/L). The clinical significance of intermediate resistance remains poorly defined. On the other hand, highly resistant strains were responsible for numerous therapeutical failures, especially in cases of meningitis. Antibiotics recommended against penicillin resistant pneumococci include cefotaxime, ceftriaxone, imipenem and in some instances vancomycin. However, penicillin-resistant pneumococci tend to present cross-resistances to all the antibotics of the beta-lactam family and could even become resistant to the last resort drugs mentioned above. Thus, in conclusion, the explosion of resistance to penicillin in pneumococci is a ubiquitous phenomenon which must be fought against by (i) a strict utilization of antibiotics, (ii) the practice of microbiological sampling of infected foci before treatment, (iii) the systematic surveillance of resistance profiles of pneumococci against antibiotics and (iv) the adequate vaccination of populations at risk. PMID- 11028186 TI - [Cutaneous signs of bacterial, fungal and parasitic infections]. PMID- 11028187 TI - The American Society of Human Genetics 50th annual meeting. October 3-7, 2000, Philadelphia, Pennsylvania, USA. Abstracts. PMID- 11028188 TI - Potential and active family caregivers: changing networks and the "sandwich generation". AB - Informal family care giving has been a traditional mainstay of care for the frail elderly. As the Baby Boomers approach retirement and old age, it is not clear that society can continue to rely on informal arrangements. The 1984 and 1994 National Long Term Care Surveys were used as sources for examining changes over a decade in the population of chronically disabled elderly, their sources of care, and the characteristics of family caregivers. The results showed that although the total number of active family caregivers declined, a constant number of primary caregivers was looking after recipients who were more severely disabled. Members of the "sandwich generation" and full-time workers maintained or even increased their participation as primary caregivers. The competing demands confronting these caregivers and the higher disability levels among care recipients probably contributed to the growing pattern of reliance on formal care, a situation that is likely to continue. PMID- 11028189 TI - Germany's long-term-care insurance: putting a social insurance model into practice. AB - A growing population of elderly has intensified the demand for long-term care (LTC) services. In response to the mounting need, Germany put into effect a LTC Insurance Act in 1995 that introduced mandatory public or private LTC insurance for the entire population of 82 million. The program was based on the organizational principles that define the German social insurance system. Those individuals in the public system and their employers each pay contributions equal to 0.85 percent of each employee's gross wages or salary. Ten percent of the population with the highest incomes have chosen the option of purchasing private long term care insurance. Provisions were made for uniform eligibility criteria, benefits based on level of care needs, cost containment, and quality assurance. Over the first four years of its operation, the system has proved financially sound and has expanded access to organized LTC services. The German system thus may serve as an example for other countries that are planning to initiate social LTC insurance systems in other nations. PMID- 11028190 TI - The Children's Health Insurance Program: expanding the framework to evaluate state goals and performance. AB - A comprehensive framework was devised to evaluate the State Children's Health Insurance Program (SCHIP) established in 1997. The framework relies on a number of potential measures and data sources for reviewing the program information recorded by states in SCHIP applications, particularly their strategic objectives and proposed performance measures. The analysis reveals that the states propose a wide range of objectives and measures and that there is considerable variation among the states. Overall, states' SCHIP plans tend to stress program enrollment and access to services but fail to emphasize the type and quality of services children receive once they are enrolled in the program. A broader conceptual framework is needed for policy makers, advocates, and researchers to make a full assessment of state goals and SCHIP performance. PMID- 11028191 TI - The health development organization: an organizational approach to achieving child health development. AB - The health development organization (HDO) is a new approach to the organization and delivery of children's health and social services. The HDO would combine the best features of vertically integrated HMOs with horizontally integrated, child focused social services and longitudinally integrated health promotion strategies. Its mandate would be to develop the health of children in a community. The impetus for creating HDOs is a growing body of evidence in chronic disease epidemiology, developmental psychopathology, early intervention research, and life course cohort studies that point to childhood as the period of life during which adult health status is determined and the opportunities for health capital formation are highest. Thus, a new kind of health care organization or framework, like the HDO, is needed to integrate a full range of critical services for promoting children's development. PMID- 11028192 TI - International Society for Autonomic Neuroscience (ISAN) Millennium Congress. London, United Kingdom. 17-21 July 2000. Abstracts. PMID- 11028193 TI - Global Congress of Gynecologic Endoscopy, 29th annual meeting of the American Association of Gynecologic Laparoscopists. Orlando, Florida, USA. November 15-19, 2000. Abstracts. PMID- 11028194 TI - ESMRMB '00. 17th Annual meeting of the European Society for Magnetic Resonance in Medicine and Biology. Paris, France, September 14-17, 2000. Abstracts. PMID- 11028195 TI - [24th Autumn session of the German Society of Cardiology and Cardiovascular Research. 21-23 September 2000, Munster. Abstracts]. PMID- 11028196 TI - [29th Congress of the German Society of Rheumatology. Aachen, 13-16 September 2000. Abstracts]. PMID- 11028198 TI - [Burkitt's lymphoma: a model for pediatric oncology]. PMID- 11028197 TI - [Lead poisoning: new role of the pediatrician]. PMID- 11028199 TI - [Kingella kingae osteoarticular infections in children. A report of a series of eight new cases]. AB - Kingella kingae is a Gram-negative bacillus which belongs to the Neisseriaceae family. Its involvement in osteoarticular infections is relatively recent. METHODS AND RESULTS: We report eight cases of Kingella kingae osteoarticular infections that have been diagnosed at the paediatric surgical centre of Rouen University Hospital since October 1995. Six boys and two girls (mean age: 30.6 months) presented with osteomyelitis in six cases and arthritis in two. Only 75% of patients had a fever at time of diagnosis. The biological findings were slightly modified. All samples were obtained from blood, bone or joint fluid. These samples were systematically inoculated into a blood culture tube. Positive Kingella kingae culture was achieved in seven local samples and in one blood culture. All children received two antibiotics via intravenous injection while waiting for the bacteriologic results. Later, the antibiotic treatment (amoxycillin) was given per os. The mean duration of treatment was 33 days. Patients were given intravenous treatment for a period of only ten days. Six patients were followed up for a period of more than 18 months and outcome was always uneventful. DISCUSSION: Kingella kingae is usually present in the nasopharyngeal mucosa and spreads in the blood due to various infections. Different types of Kingella kingae infection have been reported with a large frequency of osteoarticular infection. CONCLUSION: This type of infection does not present any unusual characteristics as compared to other osteoarticular infections. Because of its antibiotic sensitivity treatment duration could be reduced. Kingella kingae is a fragile microbe and its culture is often difficult; therefore, it is important to use blood culture tubes to inoculate joint fluid and bone samples. PMID- 11028200 TI - [Academic future of children treated for brain tumors. Single-center study of 27 children]. AB - Brain tumours constitute the most common type of solid malignancy in childhood. Despite intensive efforts developed since the mid-1970s in paediatric neuro oncology, survivors still have a wide range of sequelae leading to frequent failure in academic achievements. Very few studies have detailed the educational outcome of these children. PATIENTS AND METHODS: The study was based on a questionnaire sent to the parents of children diagnosed with a brain tumour and treated at the Centre Leon-Berard between 1987 and 1993. Children had to be under 12 years old at the time of diagnosis and with at least three years of follow-up since diagnosis. Questions focused on the child's education before diagnosis, his progress during and after treatment, the measures taken when the child experienced learning difficulties and their consequences on the child's socioprofessional integration. RESULTS: Twenty-seven responses were obtained out of 34 questionnaires. Twenty-six children were reported to experience learning difficulties. Only four children had a normal education. The main problems are associated with slowness, memory and comprehension difficulties. The main disciplines affected are mathematics, reading and spelling. Fifteen children did benefit from extra support, with large interindividual variations in the amount and the quality of this support. Half of the parents play an active role in their child's extra support. CONCLUSIONS: This study provides additional information to previous reports on progressive I.Q. decline following the treatment of a brain tumour in childhood. Learning difficulties are nearly constant and adversely influence the child's curriculum. They also affect the parents who experience questions about the future of their ideal child. The severity and complexity of these learning difficulties urge for an early multidisciplinary educational and psychological management. The main characteristics of these remedial efforts should be assessed in prospective studies. PMID- 11028201 TI - [Prenatal diagnosis of autosomal recessive polycystic kidney disease]. AB - OBJECTIVE: Prenatal ultrasonographic detection of autosomal recessive polycystic kidney disease (ARPKD) is of poor reliability, especially in early pregnancy. Molecular genetics allows earlier diagnosis, from 11 weeks of gestation; however, since only indirect diagnosis is possible--the ARPKD gene being localized on chromosome 6 but not identified--the feasibility of molecular diagnosis requires several conditions: definitive diagnosis in the index case, availability of index case and parents' DNA, genetic informativity of the family at the ARPKD locus. Results and limits of this method are analyzed, using a series of 56 requests for prenatal diagnosis. RESULTS: In eight of the 56 families ARPKD was excluded on the basis of histological (seven cases) and/or genetic (two cases) criteria. Molecular study was impossible in three families due to the lack of index case's DNA, and two other families were non-informative. Among the 43 families in which prenatal diagnosis was feasible, analysis of the haplotype of 35 fetuses issued from 29 families showed that 11 fetuses with the same haplotypes as that of the index case were affected, while 24 were not. No false positive or false negative result was reported. CONCLUSIONS: Early and reliable prenatal diagnosis of recessive polycystic kidney disease is possible in nearly 80% of affected families. PMID- 11028202 TI - [Evaluation of a prevention program against nosocomial rotavirus infections in a pediatric ward]. AB - The incidence of nosocomial rotavirus infections was evaluated by a study made in the pediatric ward in Cholet during the winter of 1993-1994. A second study was performed three years later at the same place and in similar conditions in order to evaluate the efficacy of the prevention measures taken in between. PATIENTS AND METHODS: All children below three years of age and admitted to hospital between 1 December 1996 and 15 March 1997 were included in the prospective study (348 children). Fecal specimens were collected for each patient at admission in order to search for rotavirus. Then, a second stool analysis was performed if diarrhea occurred during hospitalization or within 48 hours of discharge. These last cases were detected by a phone call. RESULTS: Thirty percent of the children had diarrhea at admission to hospital (19.3% in 1993-1994). Rotavirus was present in 11.8% of the first stool analyses (8.6% in 1993-1994). The rotavirus nosocomial infection rate has decreased from 3.7% (13 cases) in 1993-1994 to 2.9% (ten cases). The mean length of hospitalization has also decreased from 2.7 to 1.6 days. CONCLUSION: The decrease in the rotavirus nosocomial infection rate leads to enforcing the prevention measures, among which the most important seems to us to be the short length of stay. However, the study also shows the limits of prevention that are linked to the virus characteristics and the conditions of hospitalization. PMID- 11028203 TI - [Epilepsy presenting as life-threatening events in infants]. AB - OBJECTIVE: This study was aimed at assessing the frequency and features of epilepsy presenting as life-threatening events in infancy. PATIENTS AND METHODS: Fifteen cases were collected over eight years. Subtle symptoms suggestive of epilepsy were rare (9/15) in the retrospective analysis; the clinical context and cerebral imaging were occasionally contributive (5/15); interictal EEG was useful but inconstantly contributive, including when the epileptic nature of the episodes had become clinically probable (10/15). Though it had no clear relationships with epileptic episodes but contributed to mislead diagnosis, gastroesophageal reflux was frequently associated. Uncovered epilepsies were not homogeneous, ranging from severe epileptic encephalopathy to benign epilepsy of infancy. CONCLUSION: Epilepsy presenting as life-threatening events is sometimes a diagnostic challenge. When an infant develops recurrent critical episodes during a several-week follow-up, with neurological and general investigations not providing more information, the immediate success of antiepileptic treatment might support diagnosis. PMID- 11028204 TI - [Obstructive sleep apnea syndrome and hypertrophic tonsils in infants]. AB - BACKGROUND: Even if failure to thrive in infants suffering from obstructive sleep apnea syndrome (OSAS) due to hypertrophic tonsils is well documented in the literature, the surgical act is often delayed due to the lack of diagnostic evidence. CASE REPORTS: We report three cases which share the common characteristic of age of onset, tonsillar hypertrophy, growth retardation and growth catch-up after tonsillectomy. Authors emphasize the importance of clinical diagnosis as a sufficient tool in making the decision of surgery, thus avoiding unnecessary and expensive investigations. CONCLUSION: The diagnosis of OSAS in infants and children is essentially clinical, depending mainly on a history provided by the parents, laying stress on nocturnal symptoms and clinical examination. Growth retardation is frequent in this syndrome and should be systematically sought. Tonsillectomy, which is effective in relieving respiratory manifestations, also allows growth recovery. PMID- 11028205 TI - [Phrenic nerve paralysis of obstetrical origin: favorable course using continuous positive airway pressure]. AB - INTRODUCTION: Isolated diaphragmatic paralysis due to obstetrical factors is rare and therapeutic management modalities are not quite clear. CASE REPORT: A neonate born by breech delivery presented with respiratory distress due to isolated paralysis of the right hemidiaphragm. The clinical course was progressive, his condition worsening with oxygen supplementation. Continuous positive airway pressure (CPAP) delivered via a nasal cannula was started in the one-month-old child, inducing gradual improvement towards recovery at the age of two months and a half. CONCLUSION: Non-invasive nasal CPAP should be proposed for the treatment of phrenic nerve obstetrical palsy before introducing more invasive ventilation techniques. Surgical plication should be delayed until the child reaches the age of at least three months. PMID- 11028206 TI - [Role of leukotriene inhibitors in the treatment of childhood asthma]. AB - Leukotriene inhibitors are new pharmacological agents for the treatment of mild to moderate persistent asthma and exercise-induced asthma (EIA). Studies concerning their use in children remain scarce. Available data in the treatment of persistent asthma in children suggest that they could be an alternative to long-acting beta 2-agonists when asthma control cannot be obtained with inhaled steroids alone. Their main advantages are first that they are given orally once daily; second, that they do not induce tachyphylaxis to bronchoprotection against EIA, unlike long-acting beta 2-agonists. Studies specifically conducted in children are necessary to best describe their place in pediatric asthma treatment. PMID- 11028208 TI - [Radiologic case of the month. Klippel-Feil syndrome]. PMID- 11028207 TI - [Assessment of carpal bone maturation by imaging: an alternative to bone age determination or a complementary study?]. AB - Radiographic evaluation of skeletal maturation with or without automated systems relies on the comparison between the radiographic appearance of portions of a child's skeleton and the standardized appearance in a comparable population of children. Assessment of skeletal maturation using image analysis of wrist computerized-assisted tomography is a quantitative method allowing for measurements of the volume of each carpal bone, the axis of inertia, and the mean bone density. Morphological modifications of carpal bones are characterized by quantitative data relating to the length of the axis of inertia and the volume. Imaging is a potentially useful method for carpal bone assessment during growth. PMID- 11028209 TI - [Migraine, misunderstood pathology in children]. AB - Although migraine is the main chronic headache in childhood and adolescence, it remains extensively misdiagnosed. Schematically, migraine is a severe headache evolving by stereotyped attacks frequently associated with marked digestive symptoms (nausea, vomiting, abdominal pain). Throbbing pain, sensitivity to sound, and light (and sometimes odors) are frequent additional symptoms. The attack is sometimes preceded by a visual or sensory aura. Rest brings relief, and sleep often ends the attack. Childhood migraine prevalence varies between 5 and 10%. Migraine episodes are frequently triggered by several factors: emotional stress (school pressure, vexation, excitement, upset), hypoglycemia, lack or excess of sleep (weekend migraine), sensory stimulation (loud noise, bright light, strong odor, heat or cold, etc.), sympathetic stimulation (sport, physical exercise). Attack treatments must be given at an early stage, oral ibuprofen (10 mg/kg) being particularly recommended. If the oral route is not available because of nausea or vomiting, rectal or nasal routes have then to be used. Non pharmacological treatments (biofeedback and interventions combining progressive muscle relaxation) have demonstrated good efficacy as prophylactic measures. Daily prophylactic pharmacological treatments are prescribed as the second line after failure of non-pharmacological treatments. PMID- 11028210 TI - [Hearing impairment in children: early diagnosis is essential]. AB - Diagnosis of hearing impairment is possible during the first days of life. Hearing tests are noninvasive and should not be delayed when hearing loss is suspected. Among children's hearing impairments, conductive hearing loss is the most frequent; it is generally acquired and reversible. At the opposite pole, sensorineural hearing loss has more severe consequences because it is irreversible and often present from birth. Early diagnosis and treatment are necessary in all cases to prevent speech delay. In cases with sensorineural hearing impairment, hearing aid fitting, or even cochlear implantation, and intensive speech therapy will help deaf children learn speech, with the view of optimal social and professional integration. PMID- 11028211 TI - [Problems in teaching of ill children. Thoughts of a teacher in a pediatric department]. AB - The collaboration between the Department of Education and the hospital is mainly empiric. Among the questions raised by the presence of teachers in pediatric departments, one of the most important relates to the reasons why the hospital calls on the Department of Education and the purpose of their cooperation. This paper is a reflection on this question by a teacher working with sick children in a pediatric department. The relations between the teacher and his hospitalized pupil are particularly original. Teaching a sick child should not only take into account the patient's status but consider the sick child in his entirety. PMID- 11028213 TI - [Vertebral abnormalities and Williams and Beuren's syndrome]. PMID- 11028212 TI - [Filiform ankyloblepharon adnatum]. PMID- 11028214 TI - [Outcome of unclassified colitis in the ABERMAD register (inflammatory bowel disease). Effect of follow-up on incidence. Breton Association for Study and Research on Digestive System Diseases]. PMID- 11028215 TI - [Neonatal Streptococcus pneumoniae infection]. PMID- 11028216 TI - [Acute lymphoblastic leukemia revealed by lymphocytic meningitis]. PMID- 11028218 TI - In vitro iontophoretic release of lithium chloride and lidocaine hydrochloride from polymer electrolytes. AB - Ionically conducting polymers, frequently known as polymer electrolytes, are potential candidates as hosts for drugs to be delivered iontophoretically. The iontophoretic delivery of lithium or lidocaine from polymer electrolyte films through a cellophane membrane was examined using different delivery current regimes. Thin, mechanically strong, polymer electrolyte films were fabricated from poly(ethylene oxide) (PEO) with lithium chloride or lidocaine hydrochloride. Experiments showed that iontophoretic transport of both lithium chloride and lidocaine hydrochloride might be achieved from these PEO-based films. Cation transport number determinations give values for PEO-based films of about 0.4 for lithium chloride systems and 0.12 for lidocaine hydrochloride systems. The mechanism of transport from these PEO-based polymer electrolyte films allows the delivery of ionic salts such as lithium chloride and lidocaine hydrochloride to be controlled solely by current, thus providing a system that can deliver precise amounts of drug. PMID- 11028217 TI - Natural gums and modified natural gums as sustained-release carriers. AB - Although natural gums and their derivatives are used widely in pharmaceutical dosage forms, their use as biodegradable polymeric materials to deliver bioactive agents has been hampered by the synthetic materials. These natural polysaccharides do hold advantages over the synthetic polymers, generally because they are nontoxic, less expensive, and freely available. Natural gums can also be modified to have tailor-made materials for drug delivery systems and thus can compete with the synthetic biodegradable excipients available in the market. In this review, recent developments in the area of natural gums and their derivatives as carriers in the sustained release of drugs are explored. PMID- 11028219 TI - Statistical validation of reproducibility of HPLC peptide mapping for the identity of an investigational drug compound based on principal component analysis. AB - Peptide mapping is a key analytical method for studying the primary structure of proteins. The sensitivity of the peptide map to even the smallest change in the covalent structure of the protein makes it a valuable "fingerprint" for identity testing and process monitoring. We recently conducted a full method validation study of an optimized reverse-phase high-performance liquid chromatography (RP HPLC) tryptic map of a therapeutic anti-CD4 monoclonal antibody. We have used this method routinely for over a year to test production lots for clinical trials and to support bioprocess development. One of the difficulties in the validation of the peptide mapping method is the lack of proper quantitative measures of its reproducibility. A reproducibility study may include method and system precision study, ruggedness study, and robustness study. In this paper, we discuss the use of principal component analysis (PCA) to quantitate peptide maps properly using its projected scores on the reduced dimensions. This approach allowed us not only to summarize the reproducibility study properly, but also to use the method as a diagnostic tool to investigate any troubles in the reproducibility validation process. PMID- 11028220 TI - A high-performance liquid chromatography assay for yohimbine HCl analysis. AB - The analysis used yohimbine HCl solution prepared from commercially available yohimbine HCl powder. Stability-indicating high-performance liquid chromatographic (HPLC) assay procedures were established and utilized to analyze the concentration of the drug. The method proved to be a simple model since it does not contain a buffer system. The mobile phase used, a methanol:water 70:30 ratio, was similar to that suggested by the manufacturer for the storage of the column. Therefore, the solvent system saves analytical processing time since it does not require a change in the mobile phase before and after the analysis. The analytical method has been shown to be stability indicating. The assay method showed a retention time for yohimbine of 4.2 min; for caffeine, the internal standard, it was 2.3 min. The standard deviation and the coefficient of variation were under acceptable limits of 2% and were specifically 1.51% and 1.35% for within-day and between-day samples, respectively. The results showed that the degradation products obtained from stressing yohimbine HCl by heat and extremes in pH did not interfere with the yohimbine HCl peak, although the internal standard, caffeine, did show some interference due to having a retention time similar to the degradation products. PMID- 11028221 TI - Dissolution properties of piroxicam powders and capsules as a function of particle size and the agglomeration of powders. AB - The poor dissolution characteristics of relatively insoluble drugs have long been a problem to the pharmaceutical industry. An example is piroxicam, a highly potent anti-inflammatory agent. In many countries, a large number of generic piroxicam products are available to the prescriber. The aim of this study was to investigate the cause of the dissolution problems experienced by manufacturers of generic piroxicam capsules. Two raw material batches and the dissolution properties of several piroxicam capsules were studied. Differential scanning calorimetry (DSC) and X-ray powder diffraction (XRPD) results showed that the two raw material samples were identical with respect to polymorphic modification. The particles of powder 1 were smaller than those of powder 2, but the dissolution of powder 1 was much slower than that of powder 2. The dissolution results for the capsules showed a marked difference among different brands, with capsule C not meeting the USP tolerance. Adding surfactant to the dissolution medium increased the dissolution of both powder 1 and capsule C. Failure of powder 1 or capsule C to meet USP dissolution criteria could result in differences in product efficacy, as well as in potential side effects. Such observations should be taken into account along with other relevant considerations when decisions regarding the generic substitution of oral piroxicam products are made. PMID- 11028222 TI - Structure-solubility relationship and thermal decomposition of furosemide. AB - Furosemide, a high ceiling diuretic, decomposes on heating and is very sparingly soluble in water. The aim of this study was to identify the thermal decomposition product(s) of furosemide and to calculate the activation energy needed for this reaction. This was done to gain a better understanding of the unusually low water solubility of this drug. The main thermal decomposition product was identified by nuclear magnetic resonance (NMR), mass spectrometry (MS), and infrared (IR) analysis as 4-chloro-5-sulfamoylanthranilic acid (saluamine), and the activation energy, calculated from thermogravimetric analysis (TGA) measurements, for this reaction was 47.7 (+/- 1.93) kcal/mol. The experimentally measured activation energy was well below the normal 59 +/- 4 kcal/mol needed for the cleavage of the C-N bond to form saluamine. This could possibly be explained by the weakening of the C-N bond through the I-effect of the furane ring and the delocalization of the electrons of the aniline nitrogen in the chlorosulfamoyl benzoic acid entity of furosemide. This decomposition of furosemide indicates the breaking of intramolecular bonds before those of intermolecular bonds (separation of individual furosemide molecules). Strong inter- and intramolecular bonds are a probable cause for the poor water solubility of furosemide because, when some of the inter- and intramolecular bonds that form part of the hydrogen bond network disappeared, as in the structurally related decomposition product saluamine, the aqueous solubility increased. PMID- 11028223 TI - Preparation and in vitro evaluation of chitosan matrices cross-linked by formaldehyde vapors. AB - Rifampicin-chitosan matrices were prepared by a chemical cross-linking method to develop a sustained-release form. The effects of cross-linking agent (formaldehyde) on the drug release rate and release kinetics were investigated in this study. Moreover, the kinetics of rifampicin released from chitosan matrices exposed to formaldehyde vapors for predetermined time intervals were analyzed using Ritger and Peppas exponential equation. The in vitro release kinetics exhibited a non-Fickian transport model. Increasing the exposure time to formaldehyde vapors decreased the release rate of rifampicin from chitosan matrices as a result of formation of greater structural strength and tighter texture. PMID- 11028224 TI - New oral dosage form for elderly patients. III. Stability of trichlormethiazide in silk fibroin gel and various sugar solutions. AB - The hydrolysis of trichlormethiazide (TCM) in silk fibroin gel (SFG) prepared in various sugar solutions (such as ribose, fructose, mannose, and glucose solutions) was determined. The hydrolysis rate of TCM differed with the variety of sugars utilized in this study; that is, it decreased in the following order: ribose > fructose > mannose > glucose. To investigate the relationship between the hydrolysis rate of TCM and the physicochemical properties of the sugar molecule, the amount of unfrozen water of sugar molecules was calculated from differential scanning calorimetry (DSC). The amount of unfrozen water increased with an increase in the number of the equatorial OH groups n(e-OH) per sugar molecule that are able to hydrate favorably to the surrounding water molecules. The hydrolysis rate constant decreased with increase in n(e-OH); glucose, having a large n(e-OH) in this study could effectively stabilize TCM. PMID- 11028225 TI - Biodistribution of liposomes of terbutaline sulfate in guinea pigs. AB - A series of liposomes was prepared with various lipid (egg phosphatidyl choline [egg PC], phosphatidyl glycerol [PG], dipalmitoyl phosphatidyl choline [DPPC], distearoyl phosphatidyl choline [DSPC], dipalmitoyl phosphatidyl glycerol [DPPG], phosphatidyl ethanolamine [PE], cholesterol [CH], and stearylamine [SA]) compositions, such as egg PC:PG:CH (55:5:40), DPPC:PG:CH (55:5:40), DSPC:DSPG:CH (55:5:40) egg PC:SA:CH (55:5:40), DSPE:DSPG:CH (55:5:40) in molar ratio. Liposomal formulations were administered to guinea pigs intravenously; 3 hr after the treatment, serum samples and various organs (e.g., liver, spleen, lung) were removed and analyzed for drug concentration by a high-performance liquid chromatographic (HPLC) method. Based on the above study, a liposomal preparation with better lung specificity was selected, and the time profile of these liposomes was determined in guinea pigs. Three hours postadministration, a significant difference in blood levels was observed between free terbutaline sulfate and the various liposomal formulations. Localization of the drug in the lungs increased considerably when encapsulated drug was used, and the highest percentage localization was observed with DSPC:DSPG:CH (55:5:40) liposomes. The percentage recovery of the drug in the lungs with egg PC:CH:SA (55:40:5) liposomes did not change significantly when compared with egg PC:CH:PG (55:40:5) liposomes. To establish the time course of disposition of the liposomes, DSPC:DSPG:CH (55:5:40) liposomes were selected. Terminal half-life t1/2 of the drug in blood with free drug solution was about 12 hr, whereas with liposomes, a twofold increase in t1/2 was observed. The disposition data indicated that the clearance of the drug was delayed by 1.5 times when incorporated into liposomes. PMID- 11028226 TI - Enantiomeric separation and quantitative determination of atenolol in tablets by chiral high-performance liquid chromatography. AB - The separation and quantitative determination of atenolol isomers by chiral high performance liquid chromatography (HPLC) are described. Atenolol isomers were separated using a Chiralcel OD column (250 x 4.6 mm, 10 microns); the mobile phase was hexane-ethanol-diethylamine (75:25:0.1 v/v/v); ultraviolet detection was at 276 nm; and flow rate was 0.7 ml/min. The coefficient of variation and average recovery of (R)-isomer were 0.60% and 100.37%, respectively, for sample A and 0.69% and 100.63%, respectively, for sample B. The coefficient of variation and average recovery of (S)-isomer were 0.59% and 100.33%, respectively, for sample A and 0.63% and 99.78%, respectively, for sample B. PMID- 11028227 TI - Determination of the stability constants in alkanol/alpha-cyclodextrin mixed system. AB - The simultaneous determination of the stability constants for inclusion of alkanols with alpha-cyclodextrin (alpha-CD) was investigated in a mixed alkanol system using static head-space gas chromatography (SHSGC). The 1:1 stability constants obtained were in fair agreement with those obtained previously using other methods. The time required for determination of the stability constant was markedly reduced using this simultaneous SHSGC method. PMID- 11028228 TI - A screening technique to study the mechanical strength of gelatin formulations. AB - A semiquantitative method for measuring the mechanical strength of gelatin ribbons was demonstrated using a universal tensile testing machine (Instron, model 1122). Molten gelatin formulations comprised of acid-bone gelatin, limed hide gelatin, or their combinations were made, pored as gelatin films, and aged at 50% relative humidity (RH). Viscoelastic properties (mechanical strength) of five gelatin formulations were evaluated by determining elastic modulus, tensile strength, and ratio of tensile strength to elastic modulus of gelatin ribbons. This study demonstrated that a 3:1 ratio of acid-bone to limed-hide gelatin combination showed better viscoelastic properties than the other formulations studied. PMID- 11028229 TI - Preparation of cross-linked sodium alginate microparticles using glutaraldehyde in methanol. AB - Polymeric sodium alginate microparticles were prepared by precipitating sodium alginate in methanol, followed by cross-linking with glutaraldehyde. The extent of cross-linking was controlled by the time of exposure to glutaraldehyde. The topology of microparticles was characterized by scanning electron microscopy (SEM), which indicated smooth surfaces. The equilibrium swelling experiments were carried out in water to observe the effect of cross-linking and drug loading for better utility of microparticles. It was found that swelling decreased, but drug loading increased, with an increase in cross-linking of the matrix. PMID- 11028231 TI - Agreeableness as obstacle. AB - The author discusses excessive agreeableness as it appears in the analytic situation, the patient agreeing with all interpretations, accepting all the analyst suggests in such a way as to keep the treatment apparently ongoing and peaceful but actually semi-paralysed. This type of agreeableness is linked with what has been described in the literature under the rubric of compliance. The author aims to show the various ways in which this agreeableness is enacted in the relationship with the analyst, and then to explore further the kind of anxieties that this defensive structure tries to ward off. Material from an adolescent patient is used to describe the slow unravelling of very powerful anxieties about being invaded and taken over by her object, anxieties that lead to her having no life and no mind of her own. It is suggested that such deeply paranoid anxieties may unconsciously be present in other patients of this compliant, 'agreeable' type. PMID- 11028230 TI - The influence of diluent on the release of theophylline from hydrophilic matrix tablets. AB - The role of beta-cyclodextrin (beta-CD) on the apparent solubility of theophylline was investigated by the solubility method. Binary systems of theophylline and beta-CD were prepared using the dry co-grinding method. Their characterization was performed by differential scanning calorimetry (DSC). The dissolution rate of theophylline and theophylline/beta-CD and dissolution studies of matrix tablets prepared from mixtures containing theophylline and ground theophylline were carried out. It can be concluded that beta-CD is related to an increase in the apparent solubility and dissolution rate of the drug, promoting improvement on the release of theophylline from matrices manufactured with hydroxypropylmethylcellulose (HPMC). This can be attributed to the amorphous state and the increased wettability of the drug. PMID- 11028233 TI - 'Mourning and melancholia': the genesis of a text and of a concept. AB - The author investigates the origins of 'Mourning and melancholia', which has been the standard work of psychoanalytic reference on mourning since its publication. She notes that the existence of this paper has always tended to be taken for granted and that it is therefore important to identify the foundations on which Freud developed his conception of mourning, which seem to have remained almost unexamined in the literature. The internal and external sources are discussed. The former are defined as biographical data, involving a correlation between the theme of mourning and Freud's own bereavements, as well as a possible causal link between these instances and his creative activity, while the latter comprise Freud's reading and his scientific exchanges. The external sources may be subdivided into anthropological, religious (mainly Jewish), psychiatric and psychoanalytic sources. The author points out that in his essay Freud has far less to say about mourning than about melancholia and concludes that the new view of this founding text accruing from the consideration of all the above factors shows that Freud was relatively uninterested in the normal model of mourning, especially when compared with all that flowed from his work on dreams. PMID- 11028232 TI - Hypochondria: a tentative approach. AB - The author begins his investigation of hypochondria by defining the condition and distinguishing between hypochondriacal symptoms and hypochondriacal crises, the latter being stated to arise when the defensive function of the former fails. After an enumeration of the main psychogenetic elements observed in the psychoanalytic psychotherapy of a sample population of eight patients, he concentrates on the psychodynamic and psychostructural aspects and demonstrates by means of clinical examples how conflicts of separation and dependence give rise to oral-sadistic, anal-sadistic and progressive-perverse defensive structures. Although regressive and progressive psychodynamic processes established between these structures may temporarily stabilise the psychic system to some extent, splits cause them to loom large in a hypochondriacal world full of disintegrative anxiety that is accompanied by an aggressive excitation pervaded with threatening confusion and sexual perversion, where the patient flees into quasi autistic withdrawal and tormenting self-observation. Catastrophic states resulting from earlier traumas are here fused with more mature psychic elements owing to earlier traumas. The author shows how instability in these structures, together with the failure of important ego and superego functions, may lead to a malignant regression and the consequent pathogenic reintrojection of projections. When the psyche is threatened by a dynamic of this kind, hypochondriacal symptoms may ensue as a last-ditch attempt by the patient to defend against psychic decompensation. PMID- 11028234 TI - Therapeutic alliance and collaborative interactions. AB - Since its original formulation the concept of therapeutic alliance has remained basically bound to the ego-psychology model. After a first phase of heated debate about its clinical utility and its theoretical foundation, interest in this concept has waned. Even though it refers to a relational dimension it plays a minor role in the contemporary relational trend in psychoanalysis. Starting from the premise that the concept of therapeutic alliance has made a distinctive contribution to the psychoanalytic theory of technique, the author addresses the issue of the interactive dimension involved in the regulation of the collaborative relationship, focusing on some aspects of patient-analyst interactions aimed at fostering the collaborative process. These, illustrated with clinical material, concern: (1) the use of analytic method, considered as an object shared by the patient and the analyst, (2) the existence of different levels in the collaborative relationship, (3) the analysis of the pragmatic meaning of verbal communication as a tool for constructing collaborative interactions. By shifting her attention from the symbolic content of words to the pragmatic meaning of discourse--the author highlights the continuity between fostering the collaborative relationship in the course of proper interpretive activity and dealing with it directly by non-interpretive interventions. PMID- 11028235 TI - A scandal in Salzburg or Freud's surreptitious role in the 1908 Abraham-Jung dispute. AB - The author argues that it was Freud's ambition to discover the 'caput Nili' in neuropsychology and that his discourse of anxiety informed his early dialogues with the Zurich School at the Burgholzli as he tried to persuade Bleuler and Jung, experts in dementia praecox (paranoia), to replace their toxic theory with his psychosexual theory. Although Karl Abraham adopted the libido theory, Freud's campaign to persuade Jung backfired on the eve of the First International Psychoanalytical Congress in Salzburg in 1908. Freud denied Jung's theoretical apostasy by reconfiguring an Abraham-Jung plagiarism dispute into a priority dispute and then insisted that both men collude with his narration. It is argued that Freud's narrative tyranny, his 'cover-story', recycled by Jones, occludes the significance of Salzburg from the psychoanalytic discourse. PMID- 11028237 TI - Teaching Freud to undergraduates: a case report. AB - The authors describe their experience teaching 'The Writings of Sigmund Freud' to undergraduates at Columbia College over a three-year period. The course focused on Freud's developing theory of the mind, his application of psychoanalytic theory to the study of culture, and the autobiographical and historical context in which the texts were written. The students entered the course intrigued by the controversy surrounding Freud. They immersed themselves in the intellectual task of understanding Freud's work while at the same time discovering its power to speak directly to their personal concerns. The students found in Freud a companion in the search for personal meaning in the 'great books' of the western canon. At a deeper level, they found in Freud an ally in the drama of coming-of age, experienced in terms of separation and loss, concerns about normality and sexuality, the quest for autonomy and the search for identity. While interested in Freud's ideas about infantile sexuality, for the most part, they rejected the concept of the Oedipus complex. The authors present excerpts from the students' written work and classroom discussion. Their responses are explored in relation to the developmental tasks of adolescence. PMID- 11028236 TI - Patronage in the dispute over child analysis between Melanie Klein and Anna Freud -1927-1932. AB - The author investigates the role of patrons and advocates for Melanie Klein's clinical ideas at the British Psycho-Analytical Society against the backdrop of her theoretical and technical differences with Anna Freud from 1927 to 1932. He also outlines the development of Klein and Anna Freud's theories and techniques within the nascent discipline of child psychoanalysis. The London and Viennese patrons/advocates contributed to polarising what initially were clinical differences about how to analyse pre-latency and latency-age children and which technical processes might best facilitate successful treatment. While the author speculates that a diversity of motivations and agendas may have driven the London group's support for Klein--personal and politicised enthusiasm (Jones), genuine conviction (Riviere) and attempts at theoretical rapprochement between the London and Vienna schools (Glover)--he also argues that Freud's diagnosis with cancer in 1923 and suspicion of patricidal son-successors necessitated the choice of a female successor with unquestioning loyalty to his doctrines. From 1932, when Klein's clinical authority was established, her first group of English supporters began to splinter, as she went on to become a training analyst, mentor and patron in her own right to a succeeding generation of adherents who defended her views during the Controversial Discussions. PMID- 11028239 TI - Dreams and affect: a hundred years later PMID- 11028238 TI - From symbols to flesh: the polymorphous destiny of narration. AB - The author analyses certain aspects of the narration of a generally taciturn hysterico-phobic obsessional patient as they appear in the transference relationship, pinpointing its phallic mastery and the sadistic impact of the domination over the audience/analyst that underlies this mode of discourse. She examines them in relation to Proust's 'A la recherche du temps perdu', discussing the place of perversion in analytic listening and interpretation. She then outlines some of the key structuralist and formalist views of narration as a form of syntactic structure expanded by the resolution of an enigma via a hero's ordeal, arguing that if syntactic structure exists, it consists neither of affirmation nor of negation but rather of interrogation. The author notes that what makes psychoanalytic theory radically different from other interpretive theories is the co-presence of sexuality and thought: psychoanalysis reinforces the formal description of a signifying act by the unconscious psychosexual conditions of its possibility. She then discusses the poetic narrative of Nerval and also that of Proust, which is dominated by the acting out of perverse fantasy, the resulting polyphony of various psychosexual registers, its philosophical and metaphysical impact and its relevance to the analyst's own interpretive acts when formulating stories within the countertransference relationship. PMID- 11028241 TI - Finding words: affect, behaviour and meaning in the psychoanalysis of children PMID- 11028240 TI - Affect and clinical technique PMID- 11028242 TI - Affect and power PMID- 11028243 TI - Mother and baby PMID- 11028244 TI - Sandor Ferenczi: president of the International Psychoanalytical Association (1918-1920) and founder of the International Journal of Psychoanalysis. PMID- 11028245 TI - On the devaluation of the Eitingon-Freud model of psychoanalytic education. PMID- 11028246 TI - American College of Clinical Pharmacology response to the Institute of Medicine report "To err is human: building a safer health system". The Public Policy Committee. PMID- 11028247 TI - 20th-century advances in drug therapy in oncology--Part. II. AB - Ongoing research in cancer therapy has led to the development of antineoplastic agents which target specific components of the cell cycle. In Part II of this series, we discuss agents which target the mitotic mechanism by inhibiting microtubules. Although many of these agents are being shown to have multiple effects, the Vinca alkaloids and the taxanes are known as antimitotic drugs. They are among the most important anticancer agents currently available, and because of their unique mechanisms, can be combined with a wide variety of other antineoplastic agents in a spectrum of diseases. In addition, in part II, we are discussing agents that target DNA and prevent replication and thus cell growth by inhibiting the enzymes which protect DNA during replication, the topoisomerases. These drugs, too, have unique mechanisms of action and have become major components of combination regimens. The topoisomerase I inhibitors are new drugs derived from an older parent drug, and their full possibilities are still being explored. PMID- 11028248 TI - Adverse drug reactions: a review of relevant factors. AB - We examined some of the factors that contribute to the development of adverse drug reactions (ADRs) and analyzed postmarketing ADR reports for 22 drugs. The role of metabolic-based drug-drug interaction in the development of ADRs can not be overstated. Assessment of the postmarketing ADR data for 22 drugs revealed that drugs with high potential for eliciting clinically significant ADRs are usually detected and either withdrawn from the market or placed on restricted use within the first year or two of marketing. Postmarketing data could be a useful tool for understanding the ADR profile of drugs if reporting can be adequately monitored and verified. It is hoped that early evaluation of the clinically meaningful factors such as metabolism, pharmacogenetics, and effect of physiologic and pathophysiologic states on the clinical effect of a drug during drug development would significantly reduce the incidence and severity of post marketing ADRs. PMID- 11028249 TI - A contract research organization's response to the new FDA guidances for bioequivalence/bioavailability studies for orally administered drug products. AB - The new FDA Guidance for Industry BA and BE Studies for Orally Administered Drug Products--General Considerations and Average, Population, and Individual Approaches to Establishing Bioequivalence imply significant changes in the areas of enrollment, cost, ethics, time, entry, validation applications (EVAs), and statistical and pharmacokinetic methods. The changes from three-period to two period design for food effect studies, the elimination of most steady state studies, and the analyses of only the active moiety or ingredient are welcome. However, if the current guidances are adopted, additional time will be needed for participants, and more participants will be needed, resulting in higher costs to drug developers. The PK parameters needed to assess BE and the need for replicate designs for drugs with long t1/2 are still unclear. Finally, the advantages of the aggregate property of the FDA metric versus the disaggregate criteria are challenged, and four bioequivalence criteria are proposed. PMID- 11028250 TI - Comparative inhibitory activity of rofecoxib, meloxicam, diclofenac, ibuprofen, and naproxen on COX-2 versus COX-1 in healthy volunteers. AB - Steady-state inhibitory activity of rofecoxib (Vioxx) on COX-2 versus COX-1 was compared with that of commonly used nonsteroidal anti-inflammatory drugs (NSAIDs) in 76 healthy volunteers randomized to placebo, rofecoxib 12.5 mg qd, rofecoxib 25 mg qd, diclofenac 50 mg tid, ibuprofen 800 mg tid, sodium naproxen 550 mg bid, or meloxicam 15 mg qd. All of these doses include the high end of the approved clinical dose range. Ex vivo whole-blood assays were used to determine the effect on COX-2 and COX-1 activity, respectively. Urinary prostanoids were also measured. Mean inhibition of COX-2 (measured as the weighted average inhibition [WAI] of lipopolysaccharide [LPS]-induced PGE2 generation over 8 hours on day 6 vs. baseline) was -2.4%, 66.7%, 69.2%, 77.5%, 93.9%, 71.4%, and 71.5% for placebo, rofecoxib 12.5 mg, rofecoxib 25 mg, meloxicam, diclofenac, ibuprofen, and naproxen, respectively. Corresponding values for mean inhibition of COX-1 (measured as TXB2 generation in clotting whole blood) were -5.15%, 7.98%, 6.65%, 53.3%, 49.5%, 88.7%, and 94.9%. Rofecoxib had no significant effect on urinary excretion of 11-dehydro TXB2, a COX-1-derived product. These data support the contention that rofecoxib is the only drug of the regimens tested that uniquely inhibits COX-2 without affecting COX-1. PMID- 11028251 TI - The use of time step simulations and difference equations (TSSADEQ) in modeling heparin pharmacokinetics. AB - A model in which the clearance of heparin requires the binding of heparin to a finite and regenerated pool of binding was constructed using time step simulations and difference equations (TSSADEQ). A simulation of a heparin i.v. bolus demonstrated a dose-dependent, triphasic pharmacokinetic curve with (1) an initial log-linear phase representing first-order association of heparin with binding sites, (2) an intermediate plateau phase representing constant regeneration of heparin binding sites, and (3) a terminal log-linear phase occurring when the quantity of regenerated sites exceeded the remaining heparin. Sensitivity analysis based on the literature produced estimates of the k at 1.39 to 2.77 h-1, the pool of binding sites of 50 units/kg, and the regeneration rate of 15 to 20 unit/kg/h--virtually identical to the empirically derived guidelines for the bolus size and infusion rate for unfractionated heparin. A pilot study of bolus dosing of unfractionated heparin in normal volunteers confirmed the model. PMID- 11028252 TI - Pharmacokinetics and concentration-effect analysis of intravenous RGD891, a platelet GPIIb/IIIa antagonist, using mixed-effects modeling (NONMEM). AB - RGD891 is a platelet GPIIb/IIIa receptor antagonist and potent inhibitor of platelet aggregation. This compound is biotransformed in vivo to RGD039, which also exhibits high affinity for the GPIIb/IIIa receptor. Pharmacokinetic/pharmacodynamic modeling was employed to describe the concentration-effect relationship of both compounds following the intravenous administration of RGD891 to healthy volunteers. The overall objectives of this work were to support the dose selection process for future intravenous RGD891 safety and efficacy studies. Various intravenous regimens of RGD891 were administered to healthy volunteers enrolled in three Phase I studies. Frequent plasma samples were collected at regular intervals for later measurement of RGD891 and RGD039 concentrations (validated LC/MS/MS methods). The pharmacokinetics of RGD891 and RGD039 were simultaneously analyzed by nonlinear mixed-effect modeling (NONMEM). Pharmacodynamic activity was assessed in all three studies by the degree to which ADP (20 microM)-induced platelet aggregation was inhibited. Population parameters describing the concentration-effect relationship of RGD891 and RGD039 were then generated using a modified competitive Emax-based model. RESULTS: Parent compound is by far the predominant active compound circulating in the plasma following intravenous administration of RGD891. The plasma RGD891 concentration-time data were best fit by a two compartment structural model. The fit of the basic model was improved when total body weight was introduced as a covariate for RGD891 distribution. Between subject variability in the RGD891 pharmacokinetic parameters--V1, K10, and K21- was less than 17% (coefficient of variation). Formation of the active metabolite (RGD039; Km) and its elimination (Kem) were assumed to be first-order processes (i.e., one-compartment model). The population pharmacokinetic model could only provide a rough estimate of the plasma concentration-time profile for RGD039 after administration of a given intravenous dosage regimen of RGD891 since metabolite concentrations were relatively low and highly variable. The first order rate constant describing the formation of RGD039 from RGD891 (Km) was also associated with a substantial degree of between-subject variability (44.9%). The potency of RGD891 toward the inhibition of ADP-induced platelet aggregation was described by the population IC50 value (plasma concentration yielding 50% of maximal inhibition), which ranged from 58.0 to 95.4 ng/mL, depending on the pharmacokinetic-pharmacodynamic (PK-PD) model and the data set used. The relatively low concentrations of the active metabolite achieved following intravenous administration of RGD891 did not permit independent estimation of a population IC50 value for RGD039. Therefore, its potency was fixed at 2.2-fold greater than that of the parent compound (based on previous PK-PD analyses). Intersubject variability in the IC50 values was 30%. CONCLUSIONS: Antagonism of the platelet IIb/IIIa receptors by intravenously administered RGD891 was effective in inhibiting ADP-induced platelet aggregation in a reversible and dose dependent manner. Pharmacodynamic activity was largely attributed to the parent compound and less to the active metabolite based on the relative potencies of both compounds and the plasma concentrations of each achieved following intravenous administration. Intravenous bolus plus maintenance infusion regimens resulted in rapid attainment of steady-state plasma RGD891 concentrations. This combination regimen also provided for a marked and sustained inhibition of platelet aggregation that reached 90% or greater (relative to baseline values) in the higher dose groups. The modified Emax model adequately described inhibition of platelet aggregation following a particular intravenous dosage regimen of RGD891 (within the range of doses administered in the present studies). (ABSTRACT TRUNCATED) PMID- 11028253 TI - Dose-proportional and stereospecific pharmacokinetics of methylphenidate delivered using an osmotic, controlled-release oral delivery system. AB - Methylphenidate hydrochloride (HCl) is frequently used for the treatment of attention deficit/hyperactivity disorder (ADHD). A study was conducted in healthy subjects to evaluate the dose-ranging pharmacokinetics of 18, 36, and 54 mg methylphenidate HCl delivered using an oral, osmotic, controlled-release formulation (OROS). Plasma concentrations of l-methylphenidate were 40-fold lower than those of d-methylphenidate, whereas plasma concentrations of d-alpha-phenyl 2-piperidine acetic acid (d-PPA) and l-PPA, the major metabolite of methylphenidate, were comparable. Mean AUCinf values for d-methylphenidate were 42.2, 80.9, and 120 ng.h/mL for the 18, 36, and 54 mg doses, respectively, increasing dose proportionally. AUCinf values for l-methylphenidate were only approximately 1% of d-methylphenidate (0.43, 0.96, and 1.82 ng.h/mL for the 18, 36, and 54 mg dose groups, respectively). In contrast, AUCinf values of d- and l PPA were comparable. The dose-normalized d- and l-methylphenidate plasma concentration-time profiles for the three treatment groups were superimposable. Similarly, dose-normalized plasma concentrations of d- and l-PPA were superimposable. Methylphenidate metabolism, measured as the ratio of d methylphenidate AUCinf to d-PPA AUCinf and as l-methylphenidate AUCinf to l-PPA AUCinf, was similar for the three dose groups, indicating that methylphenidate metabolism was not affected by increasing dose. OROS (methylphenidate HCl) exhibits dose-proportional and linear pharmacokinetics. PMID- 11028254 TI - An evaluation of the hemostatic effect of externally applied notoginseng and notoginseng total saponins. AB - No effective hemostatic agents are available for external use. This project was conducted to evaluate the hemostatic effects of notoginseng using a hemorrhagic rat model. Rats (n = 40) were divided into four groups, and their tails were transected 5 mm from the tip. Group 1 received no treatment (control), while the other groups received external powder applied to the wound. Group 2 received placebo (flour), group 3 received ground notoginseng, and group 4 received a saponin extract of notoginseng. The total bleeding time was determined and compared between groups. The notoginseng group had lower bleeding times (9.60 +/- 1.50 min) than the control group (19.23 +/- 4.09 min, p < 0.001) or the placebo group (15.18 +/- 2.24 min, p < 0.001). Likewise, the saponin extract group had significantly lower bleeding times (11.70 +/- 2.53 min) than the control and placebo groups (p < 0.001 for both comparisons). No differences were found between the notoginseng and the saponin extract groups (p = 0.35). Notoginseng and a saponin extract from notoginseng provide hemostatic effects when applied externally. PMID- 11028255 TI - Autogenic-feedback training exercise is superior to promethazine for control of motion sickness symptoms. AB - Motion sickness symptoms affect approximately 50% of the crew during space travel and are commonly treated with intramuscular injections of promethazine. The purpose of this paper is to compare the effectiveness of three treatments for motion sickness: intramuscular injections (i.m.) of promethazine, a physiological training method (autogenic-feedback training exercise [AFTE]), and a no-treatment control. An earlier study tested the effects of promethazine on cognitive and psychomotor performance and motion sickness tolerance in a rotating chair. For the present paper, motion sickness tolerance, symptom reports, and physiological responses of these subjects were compared to matched subjects selected from an existing database who received either AFTE or no treatment. Three groups of 11 men, between the ages of 33 and 40 years, were matched on the number of rotations tolerated during their initial rotating-chair motion sickness test. The motion sickness test procedures and the 7-day interval between tests were the same for all subjects. The drug group was tested under four treatment conditions: baseline (no injections), a 25 mg dose of promethazine, a 50 mg dose of promethazine, and a placebo of sterile saline. AFTE subjects were given four 30-minute AFTE sessions before their second, third, and fourth motion sickness tests (6 hours total). The no-treatment control subjects were only given the four rotating-chair tests. Motion sickness tolerance was significantly increased after 4 hours of AFTE when compared to either 25 mg (p < 0.00003) or 50 mg (p < 0.00001) of promethazine. The control and promethazine groups did not differ. AFTE subjects reported fewer or no symptoms at higher rotational velocities than subjects in the control or promethazine groups. The primary physiological effect of promethazine was an inhibition of skin conductance level. The AFTE group showed significantly less heart rate and skin conductance variability during motion sickness tests administered after training. PMID- 11028256 TI - The relationship between terazosin dose and blood pressure response in hypertensive patients. AB - A double-blind, randomized, placebo-controlled, multicenter study was conducted to describe the dose-response curve for terazosin on blood pressure. A total of 128 patients with mild to moderate essential hypertension (supine diastolic blood pressure, 100 to 114 mmHg) participated in the study. The study consisted of a 4 week single-blind placebo lead-in period and a 14-week double-blind treatment period. Patients were randomized in equal numbers to four parallel treatment groups: terazosin 1, 2, and 5 mg; terazosin 2, 5, and 10 mg; terazosin 20, 40, and 80 mg; and placebo. The 24-hour ambulatory blood pressure measurements were performed at the end of the placebo lead-in period and at the end of each 4-week fixed-dose period. The nonlinear, mixed-effect model computer program was used to analyze the dose-response relationship. There was a strong dose-response relationship between fall in blood pressure and the 1 to 10 mg terazosin dose, as well as a plateauing of response for terazosin doses above 10 mg. The maximum antihypertensive response (Emax) to terazosin was 10.7 mmHg for systolic blood pressure and 8.0 mmHg for diastolic blood pressure. The daily dose of terazosin, which produced 50% of the maximum response (ED50), was 3.0 mg for systolic blood pressure and 1.5 mg for diastolic blood pressure. The results of this study suggest that although some patients may benefit from terazosin doses of greater than 10 mg, doses up to 10 mg will maximize therapeutic benefit for most patients, with acceptable side effects. PMID- 11028257 TI - Doxycycline-induced hypoglycemia. PMID- 11028258 TI - The ELISA guidebook. PMID- 11028259 TI - A review of the technical aspects of drug nebulization. AB - Nebulizers are widely used for the inhalation of drug solutions in a variety of respiratory diseases. The efficacy of nebulizer therapy is influenced by a great number of factors, including the design of the device and the characteristics of the drug solution. Incorrect cleaning, maintenance and disinfection procedures may change the nebulizer performance in time, whereas patient factors can influence the lung deposition of the generated aerosol. In this review the technical aspects of nebulization of drug solutions will be discussed. Two main parameters are generally used to evaluate the performance of nebulizers: the droplet size distribution of the aerosol and the drug output rate. The droplet size distribution and the drug output rate are basically determined by the design and user conditions of the nebulizer. A higher gas flow of the compressor in a jet nebulizer or a higher vibration frequency of the piezo electric crystal in an ultrasonic nebulizer, decreases the droplet size. The choice of the type of nebulizer for nebulization of a certain drug solution may initially be based on laboratory evaluation. The major part of the mass or volume distribution should preferably correspond with aerodynamic particle diameters in the range of 1 to 5 micrometer. The intended drug output must be realized within a reasonable nebulization time (less than 30 min). From the drug output only a minor fraction will be deposited in the lung. The relation between in vitro and in vivo deposition is only partly understood and to date it has not been possible to predict drug delivery only from in vitro studies on nebulizers. Therefore, studies in patients should be performed before a drug solution for nebulization can be recommended for clinical practice. The mechanical properties of nebulizers are likely to change during use. An average utilization time of nebulizers is not available. Therefore, the performance of nebulizers should be checked periodically. Patient compliance in nebulizer therapy is relatively low. This is partly due to the fact that, at present, drug solutions for nebulizers cannot be administered efficiently within a short period of time. More efficient systems should be developed. If possible, nebulizers should be substituted to more efficient systems, e.g. dry powder inhalers or metered dose inhalers. PMID- 11028260 TI - Cost analysis of neuromuscular blocking agents in the operating room: cisatracurium, atracurium, vecuronium and rocuronium. AB - Cisatracurium (C), Atracurium (A), Rocuronium (R) and Vecuronium (V) are four neuromuscular blockers (NMB) used in the operating room with similar efficacy, defined as adequate muscle relaxation, but different pharmacokinetics. C and A have organ-independent elimination, A is associated with histamine release and R has a shorter onset time. The objective of this study was to economically compare these four NMB from the hospital point of view in order to facilitate drug selection. A cost analysis was performed. Only direct costs were considered and data were collected through a retrospective chart review. A total of 151 patient charts were randomly selected. Differences between patients receiving one of the four NMB were evaluated by ANOVA or Kruskal-Wallis tests. Then a multiple linear regression analysis was conducted. In the chart review, no significant difference was observed between the four groups of patients in age, weight or surgery duration (p > 0.05). Multiple regression analysis revealed that atracurium was on average PTA 237 (1 Euro = PTA 166) cheaper per surgery than any other NMB after adjusting for other factors (p < 0.01) and there is no significant difference in cost between the other three NMBs (p > 0.1). We recommend the use of rocuronium when a quick onset is needed and the patient does not have hepatic failure, cisatracurium when a haemodynamic instability is possible and atracurium in the remaining cases. If just one NMB can be included in the drug formulary we would select cisatracurium due to its pharmacological advantages over atracurium with a small increment in cost. PMID- 11028261 TI - Evaluation of physicochemical incompatibilities during parenteral drug administration in a paediatric intensive care unit. AB - Patients in paediatric intensive care units (PICU) often receive numerous medications by the parenteral route. Frequently two or more drugs are delivered simultaneously through the same line and the risk of physicochemical incompatibilities is thus important. The objectives of this study were 1) to identify prospectively the combinations of injectable drugs administered in the PICU of our university hospital and 2) to analyze them according to information found in the literature. The data were collected by a pharmacist over a 30-day period and classified in three categories: compatible, incompatible and undocumented. Nineteen patients were included in the study with a median age of 3.2 years. The mean number (+/- SD) of injectable drugs per patient and per day was 6.5 (+/- 2.8), for a total of 26 drugs and 7 solutes. 64 combinations of drugs were observed with 2 (31.3%), 3 (45.3%), 4 (10.9%) or 5 (12.5%) drugs. 81 drug-drug and 94 drug-solute combinations were recorded. Among these, 151 (86.3%) were compatible, 6 (3.4%) incompatible and 18 (10.3%) undocumented. The incompatibilities included furosemide (Lasix), a drug in alkaline solution and Vamina-Glucose, a total parenteral nutrition solution. No clinical consequences resulting from drug incompatibilities were shown in this study. We suggest that in vitro compatibility tests on standard drug combinations, as well as a training program for nurses on drug incompatibility problems would sensitively increase the security of parenteral drug administration. PMID- 11028262 TI - Antidepressants self-poisoning and ICU admissions in a university hospital in The Netherlands. AB - OBJECTIVES: Many overdosed patients are admitted to an ICU. Antidepressants are frequently used. We examined clinical end-points of toxicity recorded during admission to our ICU of all antidepressants used in overdose. DESIGN: Single centre; retrospective analysis, 5 consecutive years (1994-1998). SETTING: Intensive and Respiratory Care Unit, Groningen University Hospital. PARTICIPANTS: 86 patients admitted to the ICU because of antidepressant self-poisoning- database of 258 consecutively admitted patients with (auto-)intoxication. RESULTS: Significantly more patients were intoxicated with TCAs (65) compared with SSRIs (20; p < 0.05), despite the fact that the number of prescriptions of antidepressants in the community was greater for SSRIs than for TCAs. Patients intoxicated with TCAs needed significantly more often tracheal intubation (27/65 vs 7/20; p < 0.05), and these individuals had also significantly more often tachycardia (14 vs. 3) and QRS-complex widening (19 vs. 1), compared to those with non-TCA antidepressant intoxication (p < 0.05). CONCLUSIONS: TCA self poisoning has remained the predominant cause of morbidity among patients with auto-intoxication admitted to our ICU in the previous years. The data from this ICU-population confirm previous evidence that SSRIs are safer in overdose than TCAs. This finding was not explained by more prescriptions in the community of TCAs compared with SSRIs. Physicians should be more reluctant in prescribing TCAs to depressed patients in whom the risk of self-poisoning is difficult to assess. PMID- 11028263 TI - Characteristics of current benzodiazepine users as indicators of differences in physical and mental health. AB - The relationship between characteristics of benzodiazepine exposure and health status was examined in order to investigate risk profiles of benzodiazepine users. In the only pharmacy of a Dutch community of 13,500 inhabitants, all current benzodiazepine users that presented with a benzodiazepine prescription in November 1994 were invited to participate. On the basis of the RAND-36 questionnaire, summary scores for both physical and mental health were calculated, the Physical Component Summary (PCS) and the Mental Component Summary (MCS). After dichotomization with a cut-off point indicating seriously impaired health and after the combination of the PCS and MCS, four different categories of health status could be identified. We used logistic regression to study the relation between these four different groups with respect to benzodiazepine exposure. In total a group of 360 current benzodiazepine users was studied. Results showed that almost one-third of the participants had no significant impaired health; this group was further classified as reference group. We classified three other groups: one with physical problems (31%), one with mental problems (18%), and one with a combination of the two (22%). Multivariate analysis showed differences in risk factors for an impaired health status. The group with impaired physical health was associated with self-reported indication for muscle relaxation, hypnotic use, and a high CDS (Chronic Disease Score). The group with impaired mental health was associated with more frequent consulting of a mental health care specialist and with a low sense of self-efficacy. The group with both impaired physical as well as mental health was associated with a higher incidence of widowhood, a lower sense of self-efficacy, a high CDS, using benzodiazepines more than prescribed, and reporting depression as reason for their benzodiazepine use. In particular, two groups need critical examination: a group of apparently healthy users with long-term benzodiazepine use; and a frail group with impaired physical and mental health and using a higher dose than prescribed. Patient counseling and management of these four groups can be tailored to the specific needs of each group. PMID- 11028264 TI - Evaluation of the impact of pharmacist's advice giving on the outcomes of self medication in patients suffering from dyspepsia. AB - The aim of this study was to investigate the outcomes of self-medication in patients suffering from dyspepsia by comparing changes in the Health related Quality of Life before and after self-medication of dyspeptic disorders. Another study objective was the quantitative and qualitative analysis of the pharmacist's advice-giving to patients with dyspepsia. Therefore the impact of the counselling by the pharmacist on the patient's health outcomes was surveyed and compared between study and control pharmacies. Moreover, the study analysed the influence of a special training on the services provided by the pharmacies with regard to self-medication. A beneficial effect of self-medication on the HRQoL of patients with dyspepsia on a weekly basis has been detected in the study. There is evidence that advice-giving and counselling by the pharmacists in self-medication have a measurable impact on self-medication outcomes. Moreover, the study reveals that patients value the information provided by the pharmacist. Pharmacists gathered the relevant and comprehensive information from the patients having dyspeptic symptoms and provided advice concerning OTC-drugs. Moreover, pharmacists frequently discussed the relevance of factors aggravating dyspeptic disorders such as lifestyle, drinking, smoking, and manner of nutrition with the patient. Training programs and treatment guidelines for the pharmacist seem to obtain a positive effect on his performance. The findings of the study substantiate the value of a pharmacist-controlled self-medication. The study results suggest that the quality of primary health care in self-medication would improve if pharmacists' involvement were even more intense. PMID- 11028265 TI - Distribution of pharmaceuticals--a Norwegian logistic perspective. AB - There is a general concern about rising costs of pharmaceutical expenses. One political measure is a more efficient distribution system, which can take the form of new channels for retailing. In Norway mail-order pharmacy (MOP) has been brought to the agenda due to the recently proposed law regulating pharmacies ("Apoteklov"), market developments abroad, demand for self-medication, the increase in OTC-products and advances in information technology. Mail-order pharmacy involves direct delivery of medications through postal mail to patients or those responsible for dispensing medication. With reference to the USA, mail order pharmacy has filled a niche in the market and several other countries are following. We are convinced that it is possible to maintain a high level of service quality in the sense of safety, counselling and compliance, and that there is a potential to develop a model for this distribution form in Norway. We believe that the actors in the Norwegian pharmaceutical market are better served in taking a more active role in this area and where possible initiating pilot projects in mail-order distribution. The pharmacists will continue to play an important role as a retail outlet and should, with their influence over patients, their knowledge and experience, contribute towards developing MOP to be a safe and complementary sales outlet. Developing such a solution demands the right balance between performance and quality on the one hand and efficiency on the other; two criteria, which we believe, do not contradict each other. PMID- 11028266 TI - A national database for essential drugs in South Africa. AB - In the process of drafting standard treatment guidelines for adults and children at hospital level, the Secretariat of the National Essential Drugs List Committee made use of a database designed with technical support from the School of Pharmacy, MEDUNSA. The database links the current 697 drugs on the Essential Drugs List with Standard Treatment Guidelines for over 400 conditions. It served to streamline the inclusion of different drugs and dosage forms in the various guidelines, and provided concise, updated information to other departments involved in drug procurement. From information on drug prices and morbidity, it can also be used to calculate drug consumption and cost estimates and compare them with actual figures. PMID- 11028267 TI - Hard and soft data: a semiotic point of view. PMID- 11028268 TI - Frequency of Cryptosporidium spp. as cause of human gastrointestinal disease in Switzerland and possible sources of infection. AB - Detection of Cryptosporidium parvum is not routinely done in laboratories of clinical microbiology and there is no obligation to communicate isolations of this pathogen to health authorities. For these reasons, frequency of cryptosporidiosis and sources of infection are only poorly known in Switzerland. To obtain more concise information in this field, feces from 5179 hospitalized and 1256 ambulatory patients with suspected gastrointestinal infections were screened for Cryptosporidium spp. over the period of one year in two laboratories. In toto, 13 patients with cryptosporidiosis were detected which results a frequency of 0.2%. Furthermore, it was shown by a projection that about 340 cases of cryptosporidiosis have to expected yearly in Switzerland, resulting an estimated morbidity of 4.85 cases per 100,000 persons. With regard to risk factors, the available patient data did not allow solid statistical conclusions. However, known risk factors such as immunosuppression, travelling abroad (33.3%) and contact to symptomatic persons were unquestionably demonstrated. Oysters, raw milk and cream from raw milk had to be strongly taken into consideration as food vehicles of transmission. Tap water from municipal nets has not to be considered as relevant source of sporadic infections. The obtained data indicate that cryptosporidiosis is a disease of low epidemiological significance in Switzerland. To a great extent, cryptosporidiosis could be prevented by best known measures of personal hygiene, avoiding certain raw food-stuffs and being aware of safe catering on travels. PMID- 11028269 TI - [Pericardial empyema as a rare complication of pneumococcal pneumonia]. AB - The case histories of 3 patients hospitalised for severe pneumococcal pneumonia are reported. Electrocardiography showed generalized ST-segment elevations and echocardiography revealed pericardial effusion. Pericardiocentesis was performed and analysis of the punctate fluid was consistent with empyema. The pericardial empyema was removed by percutaneous drainage in one patient and by thoracoscopy in 2. During the course of the disease, 2 patients developed constrictive pericarditis after 4 to 6 weeks, necessitating epi- and pericardectomy in one. When patients with pleuropneumonia present generalised ST-segment elevations in the ECG, the possibility of pericardial involvement should be evaluated by echocardiography. The pericardial empyema must be removed as soon as possible, ideally by thoracoscopic drainage. Percutaneous drainage often fails to evacuate the empyema completely and does not prevent recurrent effusions. Constrictive pericarditis occurs early in the course of the disease and is a serious complication. When a patient suffering from constrictive pericarditis remains symptomatic despite optimal conservative therapy, pericardectomy should be performed. However, as only one of our three patients required the procedure, it should not be routinely performed. PMID- 11028270 TI - [Diagnosis and course of myocarditis: a survey in the medical clinics of Zurich University Hospital 1980 to 1998]. AB - The clinical picture of myocarditis/myopericarditis is of importance in differential diagnosis, especially in younger patients with suspected myocardial infarction. Myocarditis/myopericarditis commonly presents with chest pain, and the diagnosis is usually established on clinical grounds. However, endomyocardial biopsy is necessary to confirm the diagnosis. We evaluated the characteristics of acute myocarditis over the years 1980-1998 in 54 patients of the Department of Medicine of the University Hospital, Zurich. Two to 6 patients per year were hospitalised with this diagnosis. In most cases the diagnosis was established by a combination of criteria, such as a preceding infection of the upper respiratory tract, thoracic pain, ST segment elevations in different precordial leads followed by T wave inversions, arrhythmias, elevation of cardiac enzymes, reversible hypokinesia by echocardiography and normal coronary arteries. At least 3 of 5 criteria were requested. In a first step we analysed retrospectively all patients with acute myocarditis/myopericarditis in the years 1980-1993. Among 30 cases of acute myocarditis/myopericarditis the following causes could be identified: one influenza B, one Toxoplasma gondii infection, 2 Epstein-Barr infections and one bacterial myocarditis with gram-negative rods. The aetiology of the other 25 cases remained unknown. The majority of myocarditis/myopericarditis healed without complications. One patient with Epstein-Barr myocarditis and one with Toxoplasma gondii infection died. Two patients developed dilated cardiomyopathy. In a second phase we analysed prospectively all cases with acute myocarditis/myopericarditis over the period 1994-1998: 24 patients with acute myocarditis/myopericarditis were hospitalised. At that time coronary angiography and endomyocardial biopsies were performed more frequently. We found 2 patients with giant cell myocarditis and 2 with Toxoplasma gondii infection and HIV, all of whom died. In addition, there were 2 patients with eosinophilic myocarditis, one with Lyme carditis, one with Epstein-Barr myocarditis, one with myopericarditis after Campylobacter enteritis and one histologically proven myocarditis after pneumonia with Haemophilus influenzae. The aetiology of the remaining 13 cases with myocarditis/myopericarditis could not be established. Three patients with probable viral myocarditis developed cardiogenic shock requiring intraaortic balloon pump, and fully recovered. The patient with Lyme carditis manifested with total atrioventricular block and was treated with a temporary pacemaker. One patient with lymphocytic myocarditis required heart transplantation because of terminal heart failure and one female patient with histologically proven diffuse lympho-monocytic myocarditis died of cardiogenic shock. All the other cases healed without complications. Serologies are of little diagnostic value and should be restricted to serologies with therapeutic implications. We believe that the apparent increase in myocarditis/myopericarditis in recent years is a result of better diagnostic tools, such as more specific cardiac enzyme tests, coronary angiography and endomyocardial biopsies. In most cases the therapy remains symptomatic. In elected, severe cases steroids and other immunosuppressive drugs are sometimes used. PMID- 11028271 TI - [Aging skin: physiological bases, preventive measures and therapeutic modalities]. AB - The average lifespan has increased considerably in our society. Since the skin represents the most visible organ of ageing, there is increasing interest in the physiology and treatment of wrinkles, elastosis and senile xerosis. Cutaneous ageing is a complex phenomenon consisting of genetically determined intrinsic ageing and extrinsic ageing, the latter due to sun exposure, cigarette smoking and exposure to irritants. A number of biological changes can be found during the cutaneous ageing process, including decrease of epidermal, dermal and subcutaneous cellular components and changes in the immune system. Treatment modalities include the use of emollients in the treatment of senile xerosis, and topical retinoic and glycolic acid preparations, chemical peels, botulinum, collagen and hyaluronic acid injections, dermabrasion, CO2, and Nd:Yag laser resurfacing in the treatment of wrinkles. PMID- 11028272 TI - Invasive allergic fungal sinusitis. PMID- 11028273 TI - [Aging and employment]. PMID- 11028274 TI - [Demographic change--a challenge for all involved in the labor market]. AB - The labor market and the whole economy will be confronted with the aging of the labor force in the near future. This can but must not necessarily become a problem for future innovation processes in companies. It depends on the ability of companies and their staff to abolish the common, but wrong prejustice that older worker are less innovative because of their age. PMID- 11028276 TI - [Active measures for promoting the employment of aging workers--report of an inclusive European Union Specialty Congress in Turku, Finland]. AB - This article summarizes the most important results of the Turku Conference on active strategies for an aging workforce, which took place in Turku/Finland in 1999. About 140 experts from all EU-member states participated. The article can be seen as a contribution to the current change in the debate on labor market policy concerning older workers in Germany. PMID- 11028275 TI - [Workload and health of older employees in service occupations]. AB - Presented are the results of a representative survey of the German work force (N = 34,343) regarding work load and work-related diseases. The study comprises females and males aged 45 years and older, who are occupied in the service sector (N = 8310). The findings show that older workers in this economic sector have to bear considerable physical and mental work loads that are often higher than those for the total labor force. The results presented point to the need to encourage and practice a more comprehensive health promotion for older workers in the service sector. PMID- 11028277 TI - New for old--recent developments in public policies towards older workers in the United Kingdom. AB - This article reviews recent developments in public policies towards older workers in the United Kingdom. In the last decade, against a background of population ageing, tightening labour markets and a recognition that older workers face barriers in the labour market, successive UK governments have begun to implement policies aimed at increasing labour force participation rates among this group. These have included education campaigns encouraging the recruitment and retention of older workers by firms and assistance and guidance aimed at helping older workers to obtain training and to make decisions about returning to work. The impact of these policies is discussed and proposals for developments in policies are set out. PMID- 11028278 TI - [Part-time employment for the elderly in the North Rhine-Westphalia metal and electronics industry]. AB - A postal survey targeted at 1000 firms in the metal- and electronic working industry of Northrhine Westfalia was carried out by Sozialforschungsstelle Dortmund und Institute of gerontology to analyze the implementation of the collective agreement concerning partial retirement. As the results show, firms in Northrhine Westfalia are willing to accept the collective agreement and use partial retirement though there are still problems left to solve. Firms are further using partial retirement as an instrument of early retirement. Therefore partial retirement is created as a so-called "blocked model". PMID- 11028280 TI - [Continued training of elderly workers with reference to changes in the organization of continuing vocational training]. AB - In the following contribution the under-proportional participation rate of older workers in vocational training is analyzed. Regarding the changes concerning the organization of vocational training to more job-integrated, self-directed and computer-based learning the consequences for training opportunities, motivation to learn and performance possibilities as well as institutional basic conditions to optimize the participation of elderly workers in further training are discussed. PMID- 11028279 TI - [Gainful employment and women in the 2nd half of life]. AB - The present study analyses gender-specific risks for women in the second half of their career, especially their labour force participation rate, the degree and quality of their vocational qualification, the risk of becoming unemployed, and their transition to retirement. The analysed data show that women obviously are at higher risks to participate in the labourforce throughout their biography. Therefore, strategies to improve the working conditions of female elderly workers should not only consist of age-specific policies, but of general practices in human resource development. PMID- 11028281 TI - [Utilization of medical services and medication intake of patients over 60 in Germany--health related, social structure related, socio-demographic and subjective factors]. AB - In a community sample of 394 elderly aged 61 years and older from East and West Germany, diseases, contacts with general practitioners and specialists, the use of medicine, attitudes regarding health and illness, the subjective health, psychic problems, social support, social integration, social burden, and socio demographic variables were assessed. Based on these data the determinants for the contact of physicians and the use of medicine were analyzed. The results confirmed the frequency of multimorbidity in the elderly; on average we found three different diseases at the same time for each person. In nearly 10% of the sample we found seven diagnoses existing at the same time. 88% had contact with a general practitioner at least once a year, 97% had contact either with a general practitioner or with a specialist once a year. 55.8% took at least one medicine each day. The number of diseases existing at the same time was the most determining variable for the contact of physicians and the use of medicine. Furthermore, the elderly had more contact with physicians and took more medicine if they thought they were susceptible to diseases in a high degree, and if they rated their own health as poor. Fewer contacts with physicians and a lower use of medicine were found in those elderly that rated health behavior as little useful, that had low control beliefs regarding their own health, and that experienced only a low degree of health-related limitations in their everyday life. Furthermore, we found a higher use of medicine if there was little social support. There were no significant age-related or sex-related differences regarding the contact of practitioners or the use of medicine. PMID- 11028282 TI - [Dementia patients in general practice--results of a survey]. AB - In line with the current demographic trends, the number of elderly suffering from dementia is increasing. GPs have a key position in dealing with these patients at the primary care level. Different aspects of care for patients suffering from dementia provided in general practice in Germany were assessed by means of a questionnaire (n = 563). The results highlight the role of GPs in the delivery of medical and psychosocial care. However, diagnosing dementia seems challenging and co-operation could be improved. Three fourths of the GPs diagnose vascular dementia more often than Alzheimer's disease, which is contrary to what is known from epidemiological findings. One third refers patients to a specialist on a regular basis for establishing the diagnosis. Only 4% of the German GPs entertain co-operative relationships with the "Alzheimer society" and the "Brain league". Consequences for improving these for patients with dementia and their families are discussed. PMID- 11028284 TI - [Treatment of essential hypertension--an overview]. PMID- 11028283 TI - [Subjective theories on luck and quality of life--results of explorative interviews with 65- to 74-year-old probands]. AB - Following a subject-centred perspective in the research of happiness we asked N = 26 subjects aged 65 to 74 years in semistructured interviews about their subjective theories on happiness and quality of life (QOL) (subjective contents of the terms; description of a happy person; open wishes; causal attributions; control beliefs). Content analyses resulted in 28 categories that are united into 5 domains (personal resources, social resources and interaction, activity, material and environmental resources, abstract definitions). Particular aspects in content that have not yet been reported in the literature can be interpreted as notions of happiness and QOL specific to the elderly. Differences between the subjective concepts of "happiness" versus "QOL" are described. Ratings of semantic distance between the concepts "happiness", "QOL", "satisfaction", and "health" are analysed by multidimensional scaling. According to that, subjective representations of "happiness" and "QOL" vary markedly. PMID- 11028285 TI - [Large clinical intervention trial meta-analysis]. PMID- 11028286 TI - [Cost-effectiveness of hypertension control]. PMID- 11028287 TI - [Cost-benefit analysis of ambulatory blood pressure monitoring and home blood pressure measurements]. PMID- 11028288 TI - [EBM and antihypertensive treatment]. PMID- 11028289 TI - [Treatment of hypertension and QOL assessment]. PMID- 11028290 TI - [Treatments of hypertension--nonpharmacological treatments--introduction]. PMID- 11028291 TI - [Life style modification]. PMID- 11028293 TI - [Exercise therapy for hypertension]. PMID- 11028292 TI - [Dietary treatment of hypertension]. PMID- 11028294 TI - [Smoking cessation, moderation of alcohol]. PMID- 11028295 TI - [Functional food]. PMID- 11028296 TI - [Psychosomatic treatment]. PMID- 11028297 TI - [Limitations of non-pharmacological anti-hypertensive therapy]. PMID- 11028298 TI - [Classification of antihypertensive drugs]. PMID- 11028299 TI - [Thiazide diuretics and nonthiazide diuretics]. PMID- 11028300 TI - [Loop diuretics]. PMID- 11028301 TI - [Potassium sparing diuretics]. PMID- 11028302 TI - [Beta-blockers]. PMID- 11028303 TI - [Alpha beta-blocker]. PMID- 11028304 TI - [Alpha-blocker]. PMID- 11028305 TI - [Adrenergic-inhibiting drugs]. PMID- 11028306 TI - [Direct vasodilator]. PMID- 11028307 TI - [Ca antagonists]. PMID- 11028308 TI - [Angiotensin converting enzyme inhibitors]. PMID- 11028309 TI - [Losartan potassium, an angiotensin II receptor antagonist]. PMID- 11028310 TI - [Candesartan, angiotensin II receptor antagonist]. PMID- 11028311 TI - [Parenteral antihypertensive agents]. PMID- 11028312 TI - [Psychoneurotic dysfunction]. PMID- 11028313 TI - [Water, electrolytes abnormalities]. PMID- 11028314 TI - [Hyperuricaemia]. PMID- 11028315 TI - [Sexual dysfunction due to antihypertensive drugs]. PMID- 11028316 TI - [Skin diseases]. PMID- 11028317 TI - [Orthostatic hypotension]. PMID- 11028318 TI - [Anti-hypertensive agents and cancer]. PMID- 11028319 TI - [Adverse effects of antihypertensive drugs, miscellaneous]. PMID- 11028320 TI - [The interaction of antihypertensive agents with drugs for other diseases]. PMID- 11028321 TI - [Initiation and choice of antihypertensive drug treatment]. PMID- 11028322 TI - [Combination therapy in hypertension]. PMID- 11028323 TI - [Goal of antihypertensive treatment]. PMID- 11028324 TI - [Blood pressure control to prevent target organ damage]. PMID- 11028325 TI - [Effects of antihypertensive agents on ambulatory blood pressure]. PMID- 11028326 TI - [J-curve phenomenon]. PMID- 11028327 TI - [Antihypertensive therapy and compliance]. PMID- 11028328 TI - [Clinical implications of trough to peak ratio]. PMID- 11028329 TI - [Remission of hypertension and step-down of antihypertensive drug treatment]. PMID- 11028330 TI - [Resistant hypertension]. PMID- 11028331 TI - [Antihypertensive drugs and their effects on cerebral circulation]. PMID- 11028332 TI - [The effect of antihypertensive drugs on cardiac hypertrophy]. PMID- 11028333 TI - [Effects of anti-hypertensive drugs on atherosclerosis]. PMID- 11028334 TI - [Effects of antihypertensive drugs on renal microcirculation]. PMID- 11028336 TI - [Renin inhibitors]. PMID- 11028335 TI - [Development and trends in antihypertensive drugs--introduction]. PMID- 11028337 TI - [Serotoninergic receptor antagonist]. PMID- 11028338 TI - [Potassium channel opener]. PMID- 11028339 TI - [NEP inhibitor (neutral endopeptidase inhibitor)]. PMID- 11028340 TI - [T-type calcium channel antagonists]. PMID- 11028341 TI - [Endothelin receptor antagonist]. PMID- 11028342 TI - [Vasopressin receptor antagonists]. PMID- 11028343 TI - [Future antihypertensive drugs]. PMID- 11028344 TI - [Antihypertensive treatment for acute and chronic stroke]. PMID- 11028345 TI - [Cardiac hypertrophy associated with hypertension: its management]. PMID- 11028346 TI - [Treatment of hypertensive patients associated with ischemic heart disease]. PMID- 11028347 TI - [Therapeutic strategies of heart failure due to hypertension]. PMID- 11028348 TI - [Therapy for hypertension of patients with nephropathy or dialysis]. PMID- 11028349 TI - [Treatment of hypertension complicated by peripheral vascular disease]. PMID- 11028350 TI - [Treatment of hypertension with hyperlipidemia]. PMID- 11028351 TI - [Hyperuricemia in hypertension]. PMID- 11028352 TI - [Diabetes mellitus]. PMID- 11028353 TI - [Obesity and hypertension]. PMID- 11028354 TI - [Surgical treatment of hypertension]. PMID- 11028355 TI - [A new strategy of gene therapy for the treatment of hypertension]. PMID- 11028356 TI - [Hypertension in childhood and adolescents]. PMID- 11028357 TI - [Hypertension in young adults]. PMID- 11028358 TI - [Hypertension in the elderly and target-organ damage]. PMID- 11028359 TI - [Mild hypertension]. PMID- 11028360 TI - [Borderline hypertension--overview]. PMID- 11028361 TI - [Treatments for patients with borderline hypertension]. PMID- 11028362 TI - [White coat hypertension]. PMID- 11028363 TI - [Operation and anesthesia in hypertensive patients]. PMID- 11028364 TI - [Hypertensive emergency]. PMID- 11028365 TI - [Tracking of blood pressure]. PMID- 11028366 TI - [Birth weight and blood pressure: the Barker hypothesis]. PMID- 11028367 TI - [Primary and secondary prevention of hypertension]. PMID- 11028368 TI - [Dietary protein and blood pressure]. PMID- 11028369 TI - [Homocysteine and hypertension]. PMID- 11028370 TI - [Lifestyle related diseases and hypertension]. PMID- 11028371 TI - [Obesity and hypertension]. PMID- 11028372 TI - [Insulin resistance syndrome]. PMID- 11028374 TI - [Secondary hypertension--introduction]. PMID- 11028373 TI - [Insulin-sensitizing agents and hypertension]. PMID- 11028375 TI - [Epidemiology of secondary hypertension]. PMID- 11028376 TI - [Screening of secondary hypertension]. PMID- 11028377 TI - [Renal hypertension--overview]. PMID- 11028378 TI - [Acute glomerulonephritic syndrome]. PMID- 11028379 TI - [Chronic glomerulonephritis]. PMID- 11028381 TI - [Diabetic nephropathy]. PMID- 11028380 TI - [Acute and chronic renal failure]. PMID- 11028382 TI - [Tubulointerstitial nephritis]. PMID- 11028383 TI - [Hypertension after renal transplantation]. PMID- 11028384 TI - [Hypertension in dialysis patients]. PMID- 11028385 TI - [Hypertension following renal trauma]. PMID- 11028386 TI - [Renal tumor]. PMID- 11028387 TI - [Renovascular hypertension--overview]. PMID- 11028388 TI - [Renal artery stenosis caused by atherosclerosis]. PMID- 11028389 TI - [Renal artery stenosis caused by fibromuscular dysplasia]. PMID- 11028390 TI - [Renal artery stenosis caused by aortitis syndrome (Takayasu's arteritis)]. PMID- 11028391 TI - [Treatment of renovascular hypertension]. PMID- 11028392 TI - [Endocrine hypertension--introduction]. PMID- 11028393 TI - [Pituitary disease and hypertension]. PMID- 11028394 TI - [Thyroid disorder and hypertension]. PMID- 11028395 TI - [Parathyroid disease]. PMID- 11028396 TI - [Primary aldosteronism]. PMID- 11028397 TI - [Cushing syndrome]. PMID- 11028398 TI - [Pheochromocytoma]. PMID- 11028399 TI - [Juxtaglomerular cell tumor]. PMID- 11028400 TI - [Endothelin-producing tumor]. PMID- 11028401 TI - [Cardiac and aortic diseases]. PMID- 11028402 TI - [Hypertension in vasculitis syndrome and collagen diseases]. PMID- 11028403 TI - [Blood pressure and circadian variation in blood pressure in acute stroke]. PMID- 11028404 TI - [Hypertension in brain tumor and raised intracranial pressure]. PMID- 11028405 TI - [Sleep apnea syndrome and hypertension]. PMID- 11028406 TI - [Acute psychological stress associated with a disaster]. PMID- 11028407 TI - [Perioperative management of the surgical patients with hypertension]. PMID- 11028408 TI - [Polycythemia and hypertension]. PMID- 11028409 TI - [Hypertension in pregnant women]. PMID- 11028410 TI - [Oral contraceptives and hypertension]. PMID- 11028411 TI - [Hormone replacement therapy and hypertension]. PMID- 11028413 TI - [Erythropoietin-induced hypertension]. PMID- 11028412 TI - [Drugs, poisons, and food induced hypertension]. PMID- 11028414 TI - [Cyclosporine- and tacrolimus-induced hypertension]. PMID- 11028415 TI - [Non-steroidal anti-inflammatory drugs]. PMID- 11028416 TI - [Steroid hormone- and licorice-induced hypertension]. PMID- 11028417 TI - [Tricyclic antidepressants]. PMID- 11028418 TI - [Blood pressure and lead exposure]. PMID- 11028419 TI - [Drugs and chemical substances-associated hypertension]. PMID- 11028420 TI - [Genes which causes some kinds of hereditary hypertension]. PMID- 11028421 TI - [11 beta-hydroxylase deficiency]. PMID- 11028422 TI - [17 alpha-hydroxylase deficiency]. PMID- 11028423 TI - [11 beta-Hydroxysteroid dehydrogenase deficiency]. PMID- 11028424 TI - [Glucocorticoid-remediable aldosteronism (GRA)]. PMID- 11028425 TI - [Gordon syndrome]. PMID- 11028426 TI - [Liddle's syndrome]. PMID- 11028427 TI - [Hypertension with brachydactyly]. PMID- 11028428 TI - [Multiple endocrine neoplasia]. PMID- 11028429 TI - [Neurofibromatosis]. PMID- 11028430 TI - [von Hippel-Lindau disease]. PMID- 11028431 TI - [Prostacyclin synthase gene; nonsense mutation and the gene therapy]. PMID- 11028432 TI - [Experimental models of renal hypertension]. PMID- 11028433 TI - [Animal models, DOCA-salt hypertension]. PMID- 11028434 TI - [Roles of the spontaneously hypertensive rats in the genetic study of hypertension]. PMID- 11028435 TI - [Other genetically hypertensive rats]. PMID- 11028436 TI - [Transgenic models for human hypertension]. PMID- 11028437 TI - Assessing bone density in men. PMID- 11028438 TI - Heterogeneity in the growth of the axial and appendicular skeleton in boys: implications for the pathogenesis of bone fragility in men. AB - Men with spine fractures have reduced vertebral body (VB) volume and volumetric bone mineral density (vBMD). Men with hip fractures have reduced femoral neck (FN) volume and vBMD, site-specific deficits that may have their origins in growth. To describe the tempo of growth in regional bone size, bone mineral content (BMC), and vBMD, we measured bone length, periosteal and endocortical diameters, BMC, and vBMD using dual-energy X-ray absorptiometry in 184 boys aged between 7 and 17 years. Before puberty, growth was more rapid in the legs than in the trunk. During puberty, leg growth slowed while trunk length accelerated. Bone size was more advanced than BMC in all regions, being approximately 70% and approximately 35% of their predicted peaks at 7 years of age, respectively. At 16 years of age, bone size had reached its adult peak while BMC was still 10% below its predicted peak. The legs accounted for 48%, whereas the spine accounted for 10%, of the 1878 g BMC accrued between 7 and 17 years. Peripubertal growth contributed (i) 55 % of the increase in leg length but 78% of the mineral accrued and (ii) 69% of the increase in spine length but 87% of the mineral accrued. Increased metacarpal and midfemoral cortical thickness was caused by respective periosteal expansion with minimal change in the endocortical diameter. Total femur and VB vBMD increased by 30-40% while size and BMC increased by 200-300%. Thus, growth builds a bigger but only slightly denser skeleton. We speculate that effect of disease or a risk factor during growth depends on the regions maturational stage at the time of exposure. The earlier growth of a regions size than mass, and the differing growth patterns from region to region, predispose to site-specific deficits in bone size, vBMD, or both. Regions further from their peak may be more severely affected by illness than those nearer completion of growth. Bone fragility in old age is likely to have its foundations partly established during growth. PMID- 11028439 TI - Functional characterization of osteoblasts and osteoclasts from alkaline phosphatase knockout mice. AB - Tissue nonspecific alkaline phosphatase (TNAP) knockout (ko) mice manifest defects in bone mineralization that mimic the phenotypic abnormalities of infantile hypophosphatasia. In this article, we have searched for phenotypic differences between calvarial osteoblasts and osteoclasts in wild-type (wt), heterozygous and homozygous TNAP null mice. In vitro release of 45Ca from calvarial bones, with and without stimulation with parathyroid hormone (PTH), revealed no functional difference between osteoclasts from the three TNAP genotypes. Studies of primary cultures of TNAP+/+, TNAP+/-, and TNAP-/- calvarial osteoblasts revealed no differences in the rate of protein synthesis or in the expression levels of messenger RNAs (mRNAs) for osteopontin (OP), osteocalcin (OC), collagen type I, core binding factor alpha1 (Cbfa 1), N-cadherin, Smad 5, and Smad 7. Release of interleukin-6 (IL-6) from calvarial osteoblasts under basal conditions and after stimulation with PTH, tumor necrosis factor alpha (TNF alpha) or IL-1beta was similar in all genotypes. The amount of cyclic adenosine monophosphate (cAMP) accumulation also was comparable. However, although cultures of primary TNAP-/- osteoblasts were able to form cellular nodules as well as TNAP positive osteoblasts do, they lacked the ability to mineralize these nodules in vitro. Mineralization also was delayed in TNAP+/- osteoblast cultures compared with cultures of wt osteoblasts. Incubation with media supplemented with recombinant TNAP, but not with enzymatically inactive TNAP, restored mineralization in ko osteoblast cultures. Our data provide evidence that osteoblasts in TNAP null mice differentiate normally but are unable to initiate mineralization in vitro. The fact that even heterozygous osteoblasts show delayed mineralization provides a rationale for the presence of bone disease in carriers of hypophosphatasia. PMID- 11028440 TI - The bone morphogenetic protein 2 signaling mediator Smad1 participates predominantly in osteogenic and not in chondrogenic differentiation in mesenchymal progenitors C3H10T1/2. AB - The role of the bone morphogenetic protein (BMP)-signaling mediator Smad1 in osteogenic or chondrogenic differentiation was investigated in murine parental mesenchymal progenitors C3H10T1/2 and its derivatives constitutively expressing BMP-2 (C3H10T1/2-BMP-2) and, therefore, undergo BMP-mediated osteogenic/ chondrogenic development. The functions of the three Smad1 domains, that is, the N-terminal (MH1) domain, the C-terminal (MH2) domain, and the midregional proline rich linker domain, were documented and compared with full-length Smadl. We showed that expression of the MH2 domain in parental C3H10T1/2 cells was sufficient to initiate osteogenic differentiation. Interestingly, MH1 was sufficient to initiate transcription of osteogenic marker genes like the osteocalcin or parathyroid hormone/parathyroid hormone-related protein (PTH/PTHrP) receptor. However, MH1 interfered with the histologically distinct formation of osteoblast-like cells. A dominant-negative effect on MH2-mediated osteogenic development in C3H10T1/2 cells was observed by the dose-dependent trans-expression of the midregional linker domain. Importantly, in contrast to osteogenic differentiation, Smad1 and its domains do not mimic or interfere with BMP-2-dependent chondrogenic development as monitored by the inability of MH2 to give rise to histologically distinct chondrocytes in parental C3H10T1/2 cells and by the inefficiency of the MH1 or linker domain to interfere with BMP-2-mediated chondrogenic differentiation. PMID- 11028441 TI - Further evidence for genetic heterogeneity within type II autosomal dominant osteopetrosis. AB - Type II autosomal dominant osteopetrosis (ADO II) is characterized by an increased bone mass that contrasts with the high frequency of fractures. Linkage analysis performed in an extensive Danish family recently provided evidence for the mapping of an ADO II gene to an 8.5-cM region in chromosome 1p21 between microsatellite markers D1S486 and D1S2792. We recruited, phenotyped, and haplotyped 4F catheter ADO II families including 18 affected subjects and 29 unaffected subjects in order to narrow the candidate region and to search for genetic heterogeneity. ADO II diagnosis was ascertained by the observation of vertebral end plate thickening in at least 2 patients from successive generations. Linkage studies involved five microsatellite markers (D1S486, D1S206, D1S495, D1S248, and D1S2792) spanning 1p21. Haplotype analyses of two of our families clearly excluded the tested locus. The two remaining families gave poorly informative results. These results, combined with those previously reported in two American families, suggest that chromosomal region 1p21 is most likely a minor locus for ADO II. PMID- 11028442 TI - Mineral density and bone strength are dissociated in long bones of rat osteopetrotic mutations. AB - Bone mineral density (BMD) and mechanical strength generally show strong positive correlations. However, osteopetrosis is a metabolic bone disease with increased skeletal density radiographically and increased risk of fracture. We have evaluated mechanical strength and mineral density in three osteopetrotic mutations in the rat (incisors-absent [ia/ia], osteopetrosis [op/op], and toothless [tl/tl]) to test the hypothesis that reduced bone resorption in one or more of these mutations results in weaker bones in the presence of greater mineral density and skeletal mass. Peripheral quantitative computed tomography (pQCT) was used to analyze BMD and cross-sectional geometry in the tibial diaphysis and metaphysis as well as the femoral diaphysis and femoral neck. The bending breaking force of tibial and femoral midshafts was obtained using the three-point bending test and femoral neck strength was tested by axial loading. Osteopetrotic mutants were significantly smaller than their normal littermates (NLMs) in each stock. The pQCT analysis showed that BMD and bone mineral content (BMC) were higher than or equal to NLMs in all skeletal sites measured in the osteopetrotic mutants. However, the mechanical breaking force was equal to or lower than their NLMs in all sites. The cross-sectional structure of long bone shafts was markedly different in osteopetrotic mutants, having a thin cortex and a medullary area filled with primary trabecular bone. These results indicate that osteopetrotic mutations in the rat increase bone density and decrease bone strength. The tibial diaphysis was significantly weaker in tl/tl and ia/ia mutants and the tibial metaphysis showed the greatest increase in BMD in all mutants. These data are another illustration that an increased BMD does not necessarily lead to stronger bones. PMID- 11028443 TI - Intercellular adhesion molecule 1 discriminates functionally different populations of human osteoblasts: characteristic involvement of cell cycle regulators. AB - The concept of differential regulation of certain adhesion molecules on different cell subsets and their relevance to cell functions has emerged in recent years. The initial event in bone remodeling is an increase in osteoclastic bone resorption and cell adhesion between osteoclastic precursors and bone marrow stromal cells or osteoblasts is known to commit the osteoclast development. Here, we show that human osteoblasts can be divided into two subsets based on the expression of the intercellular adhesion molecule (ICAM)-1; ICAM-1+ osteoblasts highly adhered to monocytes, including osteoclast precursors, produced osteoclast differentiation factor (ODF), and induced multinuclear osteoclast-like cell formation. Anti-ODF monoclonal antibody (mAb) did not inhibit the adhesion of monocytes to osteoblastic cells, whereas anti-leukocyte function-associated antigen (LFA)-1, a receptor for ICAM-1, mAb blocked the adhesion. We thereby propose that the higher affinity adhesion via LFA-1/ICAM-1 is prerequisite for efficient function of membrane-bound ODF during osteoclast maturation. The functional characteristics of ICAM-1+ osteoblasts were emphasized further by cell cycle regulation, as manifested by (i) up-regulation of p53 and p21, (ii) reduction of activity of cyclin-dependent kinase (cdk) 6, (iii) underphosphorylation of retinoblastoma protein, (iv) increased Fas but reduced bcl-2 expression, and (v) majority of cells remained at G0/G1 phase. Furthermore, ICAM-1+ osteoblasts were induced by interleukin-1beta (IL-1beta). Taken together, we propose that the differentiation of osteoblasts to ICAM-1+ subpopulation by inflammatory cytokines plays an important role in osteoporosis, which is observed in patients with chronic inflammation, because ICAM-1+ osteoblasts can bias bone turnover to bone resorption, committing osteoclast maturation through cell adhesion with its precursor, and the majority of ICAM-1+ osteoblasts arrested at G0/G1 phase. Such regulation of cell cycle arrest also is an important determinant of the life span of cells in bone in which continuous bone remodeling maintains its homeostasis. PMID- 11028444 TI - Force-induced osteoclast apoptosis in vivo is accompanied by elevation in transforming growth factor beta and osteoprotegerin expression. AB - The mechanism controlling the disappearance of osteoclasts from bone surfaces after bone resorption in vivo is largely unknown. This is because there is no suitable experimental system to trace the final fate of osteoclasts. Here, we used an experimental model of tooth movement in rats to show that preexisting osteoclasts disappeared from the bone surface through apoptosis during a force induced rapid shift from bone resorption to formation. On the distal alveolar bone surface of the maxillary molar in growing rats, many mature osteoclasts were present. When light tensional force was applied to the bone surface through an orthodontic appliance, these preexisting osteoclasts gradually disappeared. One day after the application of force, about 24% of the osteoclasts exhibited apoptotic morphology and the proportion of apoptotic cells was increased to 41% by day 2, then decreased afterward. These changes were undetectable on the control distal alveolar bone surface, which is free from tensional force. As shown by in situ hybridization, a marked increase in transforming growth factor beta1 (TGF-beta1) and osteoprotegerin (OPG) messenger RNA (mRNA) was observed in the stretched cells on the tensioned distal bone surface, simultaneously with the loss of osteoclasts. Both of these factors are known to have a negative effect on osteoclast recruitment and survival. As early as 2 days after force application, some of these stretched cells were identified as cuboidal osteoblasts showing intense signals for both factors. Our data suggest there may be a sequential link in tensional force applied on the bone lining cells, up-regulation of TGF beta1/OPG, and disappearance of osteoclasts. PMID- 11028445 TI - Human myeloma cells promote the recruitment of osteoblast precursors: mediation by interleukin-6 and soluble interleukin-6 receptor. AB - Multiple myeloma is associated with the development of osteolytic bone disease characterized by a disruption to normal bone resorption and bone formation. Although studies have shown that myeloma cells produce factors that promote bone resorption little data are available examining the mechanism of decreased bone formation or the factors that mediate this effect. In the present study we describe a novel in vitro coculture system in which to investigate the effect of myeloma cells on osteoblast recruitment and differentiation. Under appropriate conditions mesenchymal stem cells were shown to differentiate into colonies of cells, a proportion of which show characteristics of osteoblasts, in that they express alkaline phosphatase activity and stain positively for collagen and calcium. The addition of the human myeloma cells JJN-3, RPMI-8226, or NCI-H929 to these cultures stimulated a significant increase in the total number of colonies (p < 0.005) and the proportion of osteoblastic colonies (p < 0.005). Media conditioned by these cells also were able to promote the formation of both total and osteoblastic colonies (p < 0.005). The addition of an antibody against the interleukin-6 receptor (IL-6R) blocked myeloma cell and myeloma cell-conditioned media induced osteoblast recruitment (p < 0.01). Furthermore, media conditioned by myeloma cells incubated with phorbol ester, which promotes IL-6R shedding, or a metalloproteinase inhibitor, which inhibits IL-6R shedding, were able to stimulate (p < 0.005) and inhibit osteoblast recruitment (p < 0.005), respectively. In addition, soluble IL-6R (sIL-6R) and IL-6 together, but not alone, were able to promote osteoblastic colony formation (p < 0.01). Taken together these data show that myeloma cells promote osteoblast recruitment by release of sIL-6R from myeloma cells. PMID- 11028446 TI - Interleukin-6 gene polymorphism is related to bone mineral density during and after puberty in healthy white males: a cross-sectional and longitudinal study. AB - Bone mineral density (BMD) is under strong genetic control and is the major determinant of fracture risk. The cytokine interleukin-6 (IL-6) is an important regulator of bone metabolism and is involved in mediating the effects of androgens and estrogens on bone. Recently, a G/C polymorphism in position -174 of the IL-6 gene promoter was found. We investigated this genetic polymorphism in relation to BMD during late puberty and to peak bone mass, in healthy white males. We identified the IL-6 genotypes (GG, GC, and CC) in 90 boys, age 16.9 +/- 0.3 years (mean +/- SD), using polymerase chain reaction (PCR). BMD (g/cm2) at the femoral neck, lumbar spine, and total body was measured using dual energy X ray absorptiometry. The volumetric BMD (vBMD; mg/cm3) of the lumbar spine was estimated. Differences in BMD in relation to the genotypes were calculated using analysis of variance (ANOVA). Subjects with the CC genotype had 7.9% higher BMD of the femoral neck (p = 0.03), 7.0% higher BMD of the lumbar spine (p < 0.05), and 7.6% higher vBMD of the lumbar spine (p = 0.04), compared with their GG counterparts. Using multiple regression, the IL-6 genotypes were independently related to total body BMD (CC > GG; p = 0.03), humerus BMD (CC > GG; p < 0.05), neck BMD (CC > GG; p = 0.01), spine BMD (CC > GG; p = 0.01), and spine vBMD (CC > GG; p = 0.008). At age 19.3 +/- 0.7 years (mean +/- SD; 88 men) the IL-6 genotypes were still independent predictors for total body BMD (CC > GG; p = 0.03), humerus BMD (CC > GG; p = 0.03), spine BMD (CC > GG; p = 0.02), and spine vBMD (CC > GG; p = 0.003), while the IL-6 genotypes were not related to the increase in bone density seen after 2 years. We have shown that polymorphism of the IL-6 gene is an independent predictor of BMD during late puberty and of peak bone mass in healthy white men. PMID- 11028447 TI - Association of the vitamin D receptor genotype BB with low bone density in hyperthyroidism. AB - Bone mineral density (BMD) is modulated by genetic and environmental factors or certain diseases. In several conditions such as low calcium intake, an influence of vitamin D receptor (VDR) polymorphisms on BMD has been suggested. In the present study, we investigated the relationship of Bsm I and Fok I polymorphisms of the VDR gene and BMD in patients with hyperthyroidism, a disease that often results in low BMD. Bsm I and Fok I genotypes were determined in 76 postmenopausal hyperthyroid patients and 62 healthy postmenopausal women as controls. Patients and controls were matched for age, time since menopause, and lifestyle factors and were free of estrogen medication. BMD evaluation included axial dual X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (PQCT). Low BMD was defined as -2.5 STD below the young adult mean value. Biochemical parameters investigated were thyroid hormones, osteocalcin, and 25-(OH)-vitamin D3 as well as routine laboratory data. Low BMD was found in 61% of hyperthyroid patients and in only 23% of euthyroid controls. In the group of hyperthyroid patients with low bone density, the BB genotype (VDR Bsm I polymorphisms) was significantly more frequent (39%) than in controls (13%; p = 0.003) and hyperthyroid patients with normal BMD (6%; p = 0.013). The odds ratio (OR) for low BMD in patients with BB genotype was 5.7 (95% CI, 1.7-19.1; p < 0.005) as compared with the Bb and bb genotypes and 5.5 (95% CI, 2.3-13.2; p < 0.0001) for hyperthyroidism alone. The cumulative risk for low BMD in patients with hyperthyroidism and BB genotype was 31.4 (95% CI, 3.9-256; p < 0.0003). VDR Fok I genotypes showed no significant relationship with BMD or other general or bone-specific parameters. Thus, hyperthyroidism and the genetic background of a BB genotype may promote synergistically the development of low BMD in hyperthyroid patients. Screening for the BB genotype in these patients therefore could help to identify those with particularly high risk for the development of low BMD and allow early treatment. PMID- 11028448 TI - Calvariae from fetal mice with a disrupted Igf1 gene have reduced rates of collagen synthesis but maintain responsiveness to glucocorticoids. AB - The goals of this study were to examine the role of insulin-like growth factor I (IGF-I) on bone formation and to test the hypothesis that the inhibitory effects of glucocorticoids on bone formation are independent of the IGF-I pathway. In serum-free organ cultures of 18-day fetal mouse calvariae derived from Igf1 null mice (Igf1-/-) and their wild-type (Igf1+/+) and heterozygous (Igf1+/-) littermates, we measured the incorporation of [3H]proline into collagenase digestible protein (CDP) and noncollagen protein (NCP), percent collagen synthesis (PCS), the incorporation of [3H]thymidine into DNA, and messenger RNA (mRNA) levels of osteoblast markers in the presence or absence of dexamethasone. After 24 h of culture, calvariae of all genotypes had similar levels of PCS. However, after 48-96 h of culture, PCS was significantly lower in Igf1-/- calvariae compared with Igf1+/+ calvariae. Treatment of calvariae with 100 nM of dexamethasone for 48-96 h decreased PCS in all genotypes. After 72 h of culture, [3H]thymidine incorporation was similar in all genotypes and 100 nM dexamethasone caused a significant reduction in [3H]thymidine incorporation in all genotypes. Dexamethasone at 100 nM decreased alpha1(I)-collagen (Colla1) mRNA and increased alkaline phosphatase, bone sialoprotein, and osteopontin mRNA in all genotypes after 72 h of culture. Type I IGF receptor mRNA levels were highest in Igf1-/- calvarial cultures. Dexamethasone at 100 nM increased Igf2 and type I IGF receptor mRNA levels in all genotypes. We conclude that one intact allele for Igf1 is sufficient to maintain normal rates of collagen synthesis in fetal mouse calvarial cultures. Moreover, the inhibitory effects of glucocorticoids on collagen synthesis and cell replication are at least partially independent of the IGF-I pathway in this model. PMID- 11028449 TI - Characterization of perimenopausal bone loss: a prospective study. AB - This study characterized the change in bone mass, bone markers, pituitary/gonadal hormones, vitamin D, parathyroid hormone, and anthropometric variables in a cohort of healthy women as they passed through normal menopause. We recruited 75 women > 46 years old who had premenopausal estradiol (E2) and gonadotropin levels and regular menses. During 9.5 years of observation, 54 experienced normal menopause (PM group) and 21 remained estrogen replete (ER group). Before the beginning of the menopausal drop and after its completion, the slope of bone mass on time in the PM group was 0% for the spine, -0.61% per year for the total body, and -0.45 % per year for the femoral neck. Designating these losses as "age related," there were 0, 4.88, and 3.40% losses for spine, total body bone mineral (TBBM), and femoral neck, respectively, in the 8-year period for which the data were analyzed. Across menopause, we found a sigmoid pattern of bone loss in the PM group beginning about 2-3 years before the last menses and ending about 3-4 years after the last menses. The total estrogen-deprivation bone losses were 10.50, 7.73, and 5.30% for the spine, TBBM, and femoral neck, respectively. In the ER group, we found a 0, 0.59, and 0.93% per year loss in spine, TBBM, and femoral neck, respectively. Serum osteocalcin rose 77%, serum total alkaline phosphatase rose 34%, and urinary hydroxyproline/creatinine (Hypro/Cr) ratio rose 44% in the PM group, while remaining stable in the ER group. We conclude that menopausal bone loss is a composite of loss caused by estrogen deprivation and age per se for the hip and total body, but is caused by estrogen deprivation alone for the spine. PMID- 11028450 TI - Rate of bone loss is associated with mortality in older women: a prospective study. AB - Older women with low bone density have an increased risk of fracture, cardiovascular disease, and mortality. However, it is not known whether this association is caused by ongoing bone loss or by lower bone mass earlier in life. To determine whether rate of bone loss is associated with total and cause specific mortality, we prospectively studied 6046 women aged 65 years or older who had serial bone mineral density (BMD) measurements as a part of the Study of Osteoporotic Fractures. Rates (mean +/- SD) of loss of BMD at the heel (for a mean of 5.7 years) and hip (for a mean of 3.5 years) were estimated. Cause specific mortality was ascertained from death certificates and hospital records. BMD loss at the heel was 5.9 +/- 6.0 mg/cm2 per year (1.5 +/- 1.5%) and BMD loss at the hip was 4.1 +/- 10.2 mg/cm2 per year (0.6 +/- 1.4%). During an average follow-up of 3.2 years after the second measurement of BMD, 371 deaths occurred. Each SD increase in BMD loss at the hip was associated with a 1.3-fold (95% CI, 1.1-1.4) increase in total mortality, adjusted for age, baseline BMD, diabetes, hypertension, incident fractures, smoking, physical activity, health status, weight loss, and calcium use. In particular, hip BMD loss was associated with increased mortality from coronary heart disease (relative hazard [RH] = 1.3 per SD; 95% CI, 1.0-1.8) and pulmonary diseases (RH = 1.6 per SD; 95% CI, 1.1-2.5). The findings were similar for bone loss at the heel, except there was no significant association with pulmonary mortality. These results raise the possibility that bone loss may share common etiologies with coronary and pulmonary diseases. PMID- 11028451 TI - Three-dimensional morphometry of the L6 vertebra in the ovariectomized rat model of osteoporosis: biomechanical implications. AB - This article summarizes the results of a three-dimensional study of changes in the morphology of the L6 rat vertebra at 120 days after ovariectomy (OVX), with estrogen replacement therapy used as a positive control. Synchrotron radiation microtomography was used to quantify the structural parameters defining trabecular bone architecture, while finite-element methods were used to explore the relationships between these parameters and the compressive elastic behavior of the vertebrae. There was a 22% decrease in trabecular bone volume (TBV) and a 19% decline in mean trabecular thickness (Tb.Th) with OVX. This was accompanied by a 150% increase in trabecular connectivity, a result of the perforation of trabecular plates. Finite-element analysis of the trabecular bone removed from the cortical shell showed a 37% decline in the Young's modulus in compression after OVX with no appreciable change in the estrogen-treated group. The intact vertebrae (containing its trabecular bone) exhibited a 15% decrease in modulus with OVX, but this decline lacked statistical significance. OVX-induced changes in the trabecular architecture were different from those that have been observed in the proximal tibia. This difference was a consequence of the much more platelike structure of the trabecular bone in the vertebra. PMID- 11028452 TI - Cross-sectional geometry, bone strength, and bone mass in the proximal femur in black and white postmenopausal women. AB - Osteoporosis is characterized by both a low bone mass and a disruption of the architectural arrangement of bone tissue, leading to decreased skeletal strength and increased fracture risk. Although there are well-known ethnic differences in bone mass and fracture risk, little is known about possible ethnic differences in bone structure. Therefore, we studied cross-sectional geometry in the hip in a sample of postmenopausal black and white women in order to investigate ethnic differences that might contribute to differences in bone strength and ultimately hip fracture risk. We recruited 371 postmenopausal black and white women who were entering the Women's Health Initiative (WHI) clinical trials in Detroit. Bone density measurements of the proximal femur were done by dual-energy X-ray absorptiometry (DXA) using a Hologic 1000 Plus bone densitometer. The DXA data were used for hip structure analysis, which treats the entire proximal femur as a continuous curved beam from the proximal shaft to the femoral neck. This permits the analysis of cross-sectional geometric properties in two narrow regions corresponding to thin (5 mm) cross-sectional slabs seen on edge. The results indicate significant ethnic differences in bone density, cross-sectional geometry, and dimensional variables. Specifically, the black women have a significantly higher bone density in both locations (10.1% and 4.1% for the neck and shaft, respectively); greater cross-sectional geometric properties in the neck (ranging from 6.1% to 11.6%), but a smaller endocortical diameter in the neck (3.6%). There are fewer significant differences in cross-sectional geometry in the shaft location. Our data suggest that the spatial distribution of bone is arranged in the femoral neck to resist greater loading in black women compared with white women. PMID- 11028453 TI - Associations between maternal peak bone mass and bone mass in prepubertal male and female children. AB - The aim of this study was to estimate heritability of bone density in premenopausal women, prepubertal male, and prepubertal female child pairs. We studied 291 pairs (mothers, mean age, 33 years, range 22-45 years; children, mean age, 7.92 years, range 7.32-8.92 years). Bone density and body composition were assessed by dual-energy X-ray absorptiometry. Height and weight were measured in both mother and child. Body size-adjusted heritability estimates for areal bone density (g/cm2) were all statistically significant (femoral neck, 59%; lumbar spine, 38%; total body, 41%) and were consistently and significantly higher in mother-daughter pairs (n = 105) as compared with mother-son pairs (n = 186). Heritability estimates for bone mineral apparent density (BMAD; g/cm3) were marginally lower but remained statistically significant at all sites (femoral neck, 51%; lumbar spine, 32%; total body, 38%). Maternal osteopenia was associated with significant reductions in bone mass at all sites in the children (femoral neck, 0.75 SD and p < 0.0001; lumbar spine, 0.61 SD and p < 0.0001; total body, 0.43 SD and p = 0.012). Mother-child bone areal bone density correlation coefficients and prediction of low bone mass in the child were greater (but this did not reach statistical significance) if the corresponding anatomical site in the mother was used for prediction with the exception of the total body. These data confirm that heritability of bone mass extends to prepubertal children and is gender- and possibly site-specific as well as under separate genetic control to growth. Furthermore, the strength of the mother-child association is such that bone density screening of mothers would make it possible to identify most prepubertal children at higher risk of osteoporosis in later life. PMID- 11028455 TI - More broken bones: a 4-year double cohort study of young girls with and without distal forearm fractures. AB - Predictors of childhood fractures have not been investigated previously. This study was undertaken to determine whether a previous history of forearm fracture, low bone mineral density (BMD; both areal bone mineral density [aBMD, g/cm2] and volumetric bone mineral apparent density [BMAD, g/cm3]), or anthropometry, influence fracture risk in young girls. At baseline, two cohorts of girls, aged 3 15 years, were evaluated: 100 had recently broken a forearm (group 1) and 100 were fracture free (group 2). Four years later we restudied 170 of these girls (82 from group 1 and 88 from group 2). We now report the relationships of previous fracture history, baseline BMD (measured by dual-energy X-ray absorptiometry), baseline weight, and height to risk of new fracture. More new fractures occurred in group 1 (37 fractures in 24 girls) than in group 2 (8 fractures in 7 girls; p = 0.0007). The independent predictors for occurrence of a new fracture at any skeletal site in a multivariate model adjusting for age, weight, total body aBMD, and fracture history were previous fracture (hazard ratio [HR], 3.28; 95% CI, 1.41-7.64); age (HR per 1-year increase, 0.91; 95% CI, 0.84-0.99); total body aBMD (HR per 1 SD decrease, 1.92; 95% CI, 1.31-2.81); and body weight (HR per 1 SD increase, 1.49; 95% CI, 1.06-2.08). Girls with two risk factors together had substantially greater fracture risk: previous fracture and low spinal BMAD (HR, 9.4; 95% CI, 2.8-32.0), previous fracture and high body weight (HR, 10.2; 95% CI, 2.8-37.6), or previous fracture and low total body aBMD (HR, 13.0; 95% CI, 3.9-43.1). We conclude that previous forearm fracture, low total body aBMD, low spinal BMAD, and high body weight each increase risk of new fractures within 4 years in young girls. Interventions to reduce the risk of fractures, particularly forearm fractures, in girls warrant further study. PMID- 11028454 TI - Spontaneous, but not induced, puberty permits adequate bone mass acquisition in adolescent Turner syndrome patients. AB - Lumbar L2-L4 bone mineral density (BMD) values were measured in 37 adolescent and young adult Turner syndrome patients. Nine had developed spontaneous puberty and had had regular menses since menarche (12.55 years +/- 1.17 years) to the time of BMD evaluation (14.96 years +/- 1.26 years). In the other 28, puberty was induced with increasing doses of oral ethinyl estradiol (2.5-10.0 microg/day, for 2 years) and later administration of estrogen/gestagen therapy up to the time of BMD evaluation. In 18, the adolescent group, menarche appeared at 14.68 years +/- 0.63 years and BMD was evaluated at 17.77 years +/- 0.70 years, and in the other 10, the young adult group, menarche appeared at 14.47 years +/- 0.53 years and BMD was evaluated at 20.90 years +/- 0.68 year. BMD values were in the normal range in those who had developed spontaneous puberty (Z score values, -0.24 +/- 0.22) and in the osteopenia range in those in whom puberty was induced (Z score values, -2.09 +/- 0.79 and -2.18 +/- 0.32 for the adolescent and young adult groups, respectively) p < 0.0001. Height Z score values were similar in all three groups (-3.45 +/- 0.77, -3.15 +/- 0.83, and -3.08 +/- 0.33, respectively). No significant differences in calcium intake or physical activity were found among groups. Neither the karyotype distribution nor growth hormone (GH) therapy (five in the spontaneous puberty and six in the induced puberty groups had been treated for a 3.5- to 4.4-year period) explained the differences in BMD values. Because the main difference between groups was the availability of estrogens to bone tissue from infancy to menarche and of estrogens/gestagens from then on up to the time of BMD evaluation, our results suggest that normal gonadal function from infancy to adulthood may be required for adequate bone mass peaking. Early detection of osteopenia and improvement in general measures for adequate bone mass peaking (calcium intake and physical activity) should be considered mandatory in the health care of these patients. PMID- 11028456 TI - Vertebral fractures in Beijing, China: the Beijing Osteoporosis Project. AB - Women in China have much lower risk of hip fracture than women in Europe or North America but their risk of vertebral fractures is not known. Lateral spine radiographs, hip and lumbar spine bone density, and potential risk factors for and consequences of vertebral fractures were assessed in a random sample of 402 women age 50 years or older living in Beijing, China. The prevalence of vertebral fractures, defined by vertebral morphometry, increased from 5% (95% CI, 1-9%) in 50- to 59-year olds to 37% (27-46%) among women age 80 years or older. The age standardized prevalence of vertebral fractures was 5.5% lower than found by similar methods for women in Rochester, MN, U.S.A. Each SD lower spine bone mineral density (BMD) was associated with a 2.4-fold (1.7-3.5) increased odds of having a vertebral fracture. Women with a history of heavy physical labor had a lower risk of vertebral fractures. Vertebral fractures were associated with decreased height loss and limited physical function but not chronic back pain. Women in Beijing, China have lower bone density and a slightly lower rate of vertebral fracture than white women in the United States. Low bone density and more sedentary occupations increase the risk of fracture in women living in urban China. PMID- 11028457 TI - Contribution of organic material to the ion composition of bone. AB - Studies of bone mineral ranging from cadaveric analysis to the use of high resolution ion microprobe with secondary ion mass spectroscopy (SIMS) have concluded that bone is rich in sodium and potassium relative to calcium. Exposure of bone to acid conditions either in vitro or in vivo leads to an exchange of hydrogen ions for sodium and potassium buffering the acidity of the medium or blood, respectively. Whether these monovalent ions reside within the mineral or organic phases of bone has never been determined. To determine the contribution of organic material to bone ion composition, we dissected calvariae from 4- to 6 day-old mice, removed organic material of some with hydrazine (Hydr), and prepared all bones for analysis using a high-resolution scanning ion microprobe coupled to a secondary ion mass spectrometer. We found that in non-Hydr-treated calvariae (Ctl) there was far more surface sodium and potassium than calcium (23Na/ 40Ca = 15.7 + 1.9, ratio of counts of detected secondary ions, mean + 95% CI, 39K/40Ca = 44.0 + 1.5). Removal of organic material with hydrazine (Hydr) led to a marked fall in the ratio of sodium to calcium and potassium to calcium (23Na/40Ca = 5.9 + 1.4, p < 0.025 vs. respective Ctl and 39K/40Ca = 1.1 + 1.5, p < 0.001 vs. respective Ctl). Similarly, when examining the cross-section of the calvariae there was more sodium and potassium than calcium (23Na/40Ca = 8.6 + 1.6, 39K/40Ca = 26.7 + 1.8). Treatment with Hydr again caused a marked fall in both ratios (23Na/40Ca = 0.3 + 1.6, p < 0.001 vs. respective Ctl and 39K/40Ca = 0.02 + 1.9, p < 0.001 vs. respective Ctl). Thus, within bone the organic material contains the majority of the sodium and potassium. This suggests that the organic material in bone and not the mineral itself is responsible for the acute buffering of the additional hydrogen ions during metabolic acidosis. PMID- 11028458 TI - Alendronate administration and skeletal response during chronic alcohol intake in the adolescent male rat. AB - Alendronate is an aminobisphosphonate that inhibits bone resorption in osteoporotic humans and rats but does not induce osteomalacia. Several bisphosphonates, including alendronate, also have direct positive actions on osteoblasts, bone formation, and mineralization. We studied the effects of alendronate on skeletal development in adolescent male rats during chronic alcohol intake. Four groups of age- and weight-matched male Sprague-Dawley rats (35 days of age) were fed the Lieber-DeCarli diet containing 36% of calories as EtOH (E), the EtOH diet plus 60 mg/kg alendronate (EA) every other day intraperitoneally (ip), an isocaloric diet (I), or the isocaloric diet plus 60 mg/kg alendronate (IA) every other day ip. Body weight, femur length, serum levels of osteocalcin (OC), insulin-like growth factor 1 (IGF-1), testosterone, and luteinizing hormone (LH); femur distal metaphyseal and middiaphyseal bone mineral density (BMD) and tibial metaphyseal gene expression for alpha-1-type I collagen (Col I), OC, and bone alkaline phosphatase (AP); and femur strength by four-point bending to failure were measured after 28 days of feeding and alendronate injections. Serum alcohol levels at death were 156 +/- 13 mg/dl (E) and 203 +/- 40 mg/dl (EA). Alendronate given to alcohol-fed rats increased metaphyseal BMD by more than 3-fold over rats fed alcohol alone. Alendronate given to isocaloric pair-fed rats increased metaphyseal BMD by more than 2.5-fold over rats fed the isocaloric diet alone. Cortical BMD was reduced by alcohol but was increased by alendronate. Alcohol consumption reduced serum IGF-1 levels, and alendronate increased IGF-1 levels in alcohol-fed rats. Serum OC, testosterone, and LH were unaffected by alcohol and alendronate. Quantitative dot blot hybridization using rat complementary DNA (cDNA) probes and normalization against 18S subunit ribosomal RNA (rRNA) levels revealed no changes in tibial metaphyseal gene expression for type I collagen, osteocalcin, or alkaline phosphatase. Alcohol significantly reduced the biomechanical properties of the femurs that were partially compensated by alendronate. Chronic alcohol consumption uncouples formation from ongoing resorption, and resorption is inhibited by alendronate. However, alendronate's positive effects on osteoblast-mediated mineralization during chronic alcohol consumption point to the potential use of bisphosphonates in the treatment of decreased bone formation secondary to alcohol-induced diminished osteoblast function. PMID- 11028459 TI - Concentration of bisphosphonate (incadronate) in callus area and its effects on fracture healing in rats. AB - The aim of the present study was to investigate effects of incadronate on early stages of fracture healing and to detect its concentration in callus area (Ca.Ar). Rats were injected three times per week with either two doses of incadronate (10 microg/kg and 100 microg/kg) or vehicle for 2 weeks. Femora were then fractured and fixed and animals were divided into pretreatment (P-10 and P 100) and continuous treatment (C-10 and C-100) groups. Incadronate treatment was stopped in P-10 and P-100 groups but continued in C-10 and C-100 groups. Animals were killed at 2 weeks and 4 weeks after fracture. Results showed significantly large callus, compared with the control, only in C-100 group at 4 weeks but not at 2 weeks. Both linear labeled surface (LS) and eroded surface (ES) decreased significantly in C-10 and C-100 groups at 2 weeks and 4 weeks. Osteoclast number (N.Oc) decreased significantly in C-10 and C-100 groups at 2 weeks but increased slightly at 4 weeks. However, there was no significant difference in the above parameters in P-10 and P-100 groups at 4 weeks. Apoptotic osteoclasts were observed only in the C-100 group at 4 weeks. A time-course decrease in incadronate concentration was detected in P-10 and P-100 groups whereas an increase was observed in C-10 and C-100 groups. These findings suggest that larger callus under incadronate treatment may result from the inhibition of bone resorption, histological characteristics of callus may be correlated with incadronate concentration, and metabolism of incadronate in bone may be related to the rate of bone turnover. PMID- 11028460 TI - Intravenous pamidronate for treatment of reflex sympathetic dystrophy during breast feeding. AB - A 39-year-old woman presented in the first month of pregnancy with reflex sympathetic dystrophy involving both lower legs. Symptoms became so severe that she could not walk unassisted, and the pain worsened after delivery. Radiographs showed patchy reduction in apparent density in the tarsal bones and around the ankles and knees. Uptake was increased in these areas on technetium methylene diphosphonate bone scan. Bone density (dual-energy X-ray absorptiometry) was reduced in the spine, hip, and radius. Biochemical tests were normal except for an increase in urinary excretion of the N-telopeptide cross-linking region of type I collagen (NTx). Because the patient wanted to continue breast-feeding, intravenous pamidronate was administered at monthly intervals. Breast milk was collected for 48 h after the infusion. The pain began to decrease soon after drug administration was initiated, and it was virtually gone by 6 months. NTx excretion fell by 78% and bone density increased by as much as 18.9% over the 6 month treatment interval. The baby was healthy and grew normally. Milk expressed after the first treatment was assayed for pamidronate content by high-performance liquid chromatography with fluorescence detection. None was detected (limit of quantitation, 0.4 micromol/liter). This case shows that pamidronate may be considered for treatment of lactating women. PMID- 11028461 TI - Massive tumoral calcinosis in a patient on long-term hemodialysis. PMID- 11028462 TI - Hemodynamic and clinical impact of prosthesis-patient mismatch in the aortic valve position and its prevention. AB - Prosthesis-patient mismatch is present when the effective orifice area of the inserted prosthetic valve is less than that of a normal human valve. This is a frequent problem in patients undergoing aortic valve replacement, and its main hemodynamic consequence is the generation of high transvalvular gradients through normally functioning prosthetic valves. The purposes of this report are to present an update on the concept of aortic prosthesis-patient mismatch and to review the present knowledge with regard to its impact on hemodynamic status, functional capacity, morbidity and mortality. Also, we propose a simple approach for the prevention and clinical management of this phenomenon because it can be largely avoided if certain simple factors are taken into consideration before the operation. PMID- 11028463 TI - Provisional stenting strategies: systematic overview and implications for clinical decision-making. AB - Coronary stents reduce the rates of abrupt closure, emergency coronary artery bypass graft surgery and restenosis, but do not prevent myocardial infarction or death at six months. The financial burden of increased stent use and the difficulty in managing in-stent restenosis have provided the impetus to develop provisional stenting strategies. Patients at low risk for restenosis after balloon angioplasty may not derive additional benefit from stent implantation and may be successfully managed with percutaneous transluminal coronary angioplasty (PTCA) alone. Numerous patient, lesion and procedural predictors of restenosis have been identified. Postprocedural assessment using quantitative coronary angiography, intravascular ultrasound (IVUS), coronary flow velocity reserve (CVR) or fractional flow reserve (FFR) may further enhance the ability to predict adverse outcomes after PTCA. Several studies have been performed to investigate the feasibility of provisional stenting strategies using various modalities to identify low risk patients who could be managed with PTCA alone. An optimal or "stent-like" angiographic result after PTCA is associated with favorable clinical outcomes. Preliminary results of studies using IVUS or CVR to guide provisional stenting appear promising. Angiography alone may be inadequate to identify truly low risk patients and may need to be combined with clinical factors, assessment of recoil, IVUS or physiologic indexes. Strategies that avoid unnecessary stenting in even a small proportion of patients may have large impacts on health care costs. Provisional stenting may potentially reduce costs and rates of in stent restenosis without compromising the quality of health care delivery. PMID- 11028464 TI - Outcomes 15 years after valve replacement with a mechanical versus a bioprosthetic valve: final report of the Veterans Affairs randomized trial. AB - OBJECTIVES: The goal of this study was to compare long-term survival and valve related complications between bioprosthetic and mechanical heart valves. BACKGROUND: Different heart valves may have different patient outcomes. METHODS: Five hundred seventy-five patients undergoing single aortic valve replacement (AVR) or mitral valve replacement (MVR) at 13 VA medical centers were randomized to receive a bioprosthetic or mechanical valve. RESULTS: By survival analysis at 15 years, all-cause mortality after AVR was lower with the mechanical valve versus bioprosthesis (66% vs. 79%, p = 0.02) but not after MVR. Primary valve failure occurred mainly in patients <65 years of age (bioprosthesis vs. mechanical, 26% vs. 0%, p < 0.001 for AVR and 44% vs. 4%, p = 0.0001 for MVR), and in patients > or =65 years after AVR, primary valve failure in bioprosthesis versus mechanical valve was 9 +/- 6% versus 0%, p = 0.16. Reoperation was significantly higher for bioprosthetic AVR (p = 0.004). Bleeding occurred more frequently in patients with mechanical valve. There were no statistically significant differences for other complications, including thromboembolism and all valve-related complications between the two randomized groups. CONCLUSIONS: At 15 years, patients undergoing AVR had a better survival with a mechanical valve than with a bioprosthetic valve, largely because primary valve failure was virtually absent with mechanical valve. Primary valve failure was greater with bioprosthesis, both for AVR and MVR, and occurred at a much higher rate in those aged <65 years; in those aged > or =65 years, primary valve failure after AVR was not significantly different between bioprosthesis and mechanical valve. Reoperation was more common for AVR with bioprosthesis. Thromboembolism rates were similar in the two valve prostheses, but bleeding was more common with a mechanical valve. PMID- 11028465 TI - How deadly is the "deadly quartet"? A post-CABG evaluation. AB - OBJECTIVES: The aim of the study was to determine the value of a cluster of metabolic risk factors in predicting mortality after coronary artery bypass surgery (CABG). BACKGROUND: The "deadly quartet" of metabolic risk factors (i.e., obesity, diabetes, hypertension, and hypertriglyceridemia) has been associated with coronary heart disease in healthy population studies. The expected influence of the cluster on survival in secondary prevention remains untested overall as well as by gender. METHODS: Patients with lipid profiles undergoing primary isolated CABG (n = 6,428) between 1987 and 1992 were followed a median of eight years. Cox models were used to evaluate all-cause mortality. Metabolic risk factors were incorporated as the sum of deadly quartet risk factors present in each patient (0 to 4). The role of gender as it relates to survival and metabolic risk clusters was also examined. RESULTS: The sum of deadly quartet risk factors showed a significant relationship to mortality as the hazard ratio increased from 1.64 (confidence interval [CI] = 1.34-2.01) for one risk factor to 3.95 (2.73 5.69) for four risk factors. Annualized mortality ranged from 1% per year in patients with no risk factors to 3.3% per year in patients with all four risk factors. Within gender, the hazard ratio associated with four risk factors was 2.58 for men and 13.39 for women. The expected clustering of risk factors was 8% compared to the observed clustering of 10% in men and 21% in women. CONCLUSIONS: This cohort showed risk factor clustering beyond that expected due to chance, particularly in women. Even after revascularization, survival is diminished for patients with members of a family of metabolic risk factors at the time of surgery. PMID- 11028466 TI - Diabetes and outcomes of coronary artery bypass graft surgery in patients with severe left ventricular dysfunction: results from The CABG Patch Trial database. The CABG Patch Trial Investigators and Coordinators. AB - OBJECTIVES: We examined the relationship between diabetes mellitus and outcomes after coronary artery bypass graft (CABG) surgery in patients with severe left ventricular (LV) dysfunction. BACKGROUND: Although diabetes is associated with poor outcomes after CABG surgery among unselected patients, the relationship between diabetes and mortality after CABG surgery among patients with LV dysfunction is less certain. METHODS: Using data from The CABG Patch Trial, a study of implantable cardiac defibrillator therapy, we analyzed 900 patients with ejection fraction <0.36 who underwent CABG surgery from 1990 to 1996. RESULTS: Diabetics comprised 38% of the patients, and 48% of diabetics were prescribed insulin. Diabetes was associated with hypertension, peripheral vascular disease, history of stroke, clinical heart failure and rales on physical exam. Diabetics were at higher risk for postoperative superficial sternal wound infection and renal failure. With an average follow-up time of 32 +/-16 months, actuarial all cause mortality 48 months after CABG surgery was 26% in diabetics and 24% in nondiabetics (p = 0.66, log-rank test). Diabetes was not associated with long term mortality in Cox multiple regression analyses. Actuarial re-hospitalization rates 48 months after CABG surgery were 85% in diabetics and 69% in nondiabetics (p = 0.0001, log-rank test). Diabetics had a 44% higher risk of re hospitalization for any cause (p = 0.0001) and a 24% higher risk of re-admission for cardiac causes (p < 0.05). Unexpectedly, fewer arrhythmic events were found in diabetics. CONCLUSIONS: Diabetes was not a predictor of mortality after CABG surgery among patients with LV dysfunction despite associated comorbidities. However, diabetes was associated with increased postoperative complications and re-hospitalization. PMID- 11028467 TI - Up to seven years of experience with the Ross procedure in patients >60 years of age. AB - OBJECTIVES: The objective of this study was to compare the outcome of patients >60 years of age with younger patients after the Ross procedure. BACKGROUND: Currently, the Ross procedure is performed predominantly in young patients. Main arguments against the Ross procedure in the elderly are the complexity of the operation and related risks. Experience with the Ross procedure in patients >60 years of age is scarce. METHODS: Between February 1990 and August 1998, the Ross procedure was performed in 27 patients (15 men and 12 women) >60 years of age (mean 64.2 +/- 3.1 years, range 60.5 to 70.6; group A) and in 84 patients (68 men, 12 women) <60 years of age (mean 43.8 +/- 12.4 years, range 15.2 to 59.4; group B). Echocardiography was applied at a mean follow-up of 28.4 +/- 21.0 and 25.2 +/- 21.4 months, respectively, to determine hemodynamic variables (ejection fraction, fractional shortening, stroke volume, cardiac output), cardiac dimensions and autograft and homograft valve function. RESULTS: There was one early and one late (esophageal bleeding) death in group B; the mortality rate was 0% in group A. One autograft was replaced because of a subvalvular aneurysm, and one patient was lost to follow-up (group B). There were no significant differences in cardiac dimensions, grade of insufficiencies across homografts and autografts and hemodynamic variables, except for a higher pressure gradient across the homograft in group B (maximal pressure gradient 11.3 +/- 5.6 vs. 7.7 +/- 4.6 mm Hg in group A). The median New York Heart Association functional class was I in both groups. CONCLUSIONS: Our seven years of experiences (mean follow-up 28 months) indicate that the Ross procedure may be performed in selected patients >60 years of age without increased risk for mortality or complications in experienced centers. PMID- 11028468 TI - Survival analysis and risk factors for mortality in transplantation and staged surgery for hypoplastic left heart syndrome. AB - OBJECTIVES: We compared survival in treatment strategies and determined risk factors for one-year mortality for hypoplastic left heart syndrome (HLHS) using intention-to-treat analysis. BACKGROUND: Staged revision of the native heart and transplantation as treatments for HLHS have been compared in treatment-received analyses, which can bias results. METHODS: Data on 231 infants with HLHS, born between 1989 and 1994 and intended for surgery, were collected from four pediatric cardiac surgical centers. Status at last contact for survival analysis and mortality at one year for risk factor analysis were the outcome measures. RESULTS: Survival curves showed improved survival for patients intended for transplantation over patients intended for staged surgery. One-year survival was 61% for transplantation and 42% for staged surgery (p < 0.01); five-year survival was 55% and 38%, respectively (p < 0.01). Survival curves adjusted for preoperative differences were also significantly different (p < 0.001). Waiting list mortality accounted for 63% of first-year deaths in the transplantation group. Mortality with stage 1 surgery accounted for 86% of that strategy's first year mortality. Birth weight <3 kg (odds ratio [OR] 2.4), highest creatinine > or =2 mg/dL (OR 4.7), restrictive atrial septal defect (OR 2.7) and, in staged surgery, atresia of one (OR 4.2) or both (OR 11.0) left-sided valves produced a higher risk for one-year mortality. CONCLUSIONS: Transplantation produced significantly higher survival at all ages up to seven years. Patients with atresia of one or both valves do poorly in staged surgery and have significantly higher survival with transplantation. This information may be useful in directing patients to the better strategy for them. PMID- 11028469 TI - Influence of diabetes mellitus on early and late clinical outcomes in saphenous vein graft stenting. AB - OBJECTIVES: The purpose of this study was to compare early and late clinical outcomes in diabetic and nondiabetic patients after stent implantation in saphenous vein grafts (SVG). BACKGROUND: Patients with diabetes mellitus have less favorable acute and long-term outcomes after stent implantation in native coronary arteries. The impact of diabetes on SVG stenting, however, is not known. METHODS: We studied 908 consecutive patients (1,366 SVG lesions) treated with Palmaz-Schatz stents. In-hospital and late clinical outcomes (death, Q-wave myocardial infarction and repeat revascularization rates at one year) were compared between diabetic (n = 290) and nondiabetic (n = 618) patients. RESULTS: In-hospital mortality was significantly higher in diabetic as compared with nondiabetic patients (2.2% vs. 0.3%, p = 0.003). At one-year follow-up, target lesion revascularization (TLR) was 16.6% in diabetic and 12.3% in nondiabetic patients (p = 0.03). Overall cardiac event-free survival (freedom from death, Q wave myocardial infarction and any coronary revascularization procedure) at one year was significantly lower in the diabetic (68%) compared with the nondiabetic patients (79%, p = 0.0003). By Cox regression analysis, diabetes mellitus was an independent predictor of both TLR (relative risk: 1.23; confidence interval: 0.96 to 1.58; p = 0.004) and late cardiac events (relative risk: 1.40; confidence interval: 1.05 to 1.86; p = 0.02). CONCLUSIONS: Patients with diabetes undergoing stent implantation in SVG have: 1) higher in-hospital and late mortality, 2) higher one-year TLR rates, and 3) significantly lower one-year cardiac event-free survival. Thus, diabetic patients have less favorable acute and late clinical outcomes after stent implantation in SVG lesions. PMID- 11028470 TI - Short- and long-term mortality for patients undergoing primary angioplasty for acute myocardial infarction. AB - OBJECTIVES: The goal of this study was to learn more about the risk factors and short- and long-term outcomes for primary angioplasty. BACKGROUND: Primary angioplasty (direct angioplasty without antecedent thrombolytic therapy) has been an effective alternative to thrombolytic therapy for patients with acute myocardial infarction (AMI). However, most reported studies have been compromised by small sample sizes and short observation times. METHODS: New York's coronary angioplasty registry was used to identify New York patients undergoing angioplasty within 6 h of AMI between January 1, 1993 and December 31, 1996. Statistical models were used to identify significant risk factors for in-patient and long-term survival and to estimate long-term survival for all patients as well as various subsets of patients undergoing primary angioplasty. RESULTS: The in-hospital mortality rate for all primary angioplasty patients was 5.81%. When patients in preprocedural shock (who had a mortality rate of 45%) were excluded, the in-hospital mortality rate dropped to 2.60%. Mortality rates for all primary angioplasty patients at one year, two years and three years were 9.3%, 11.3% and 12.6%, respectively. Patients treated with stent placement did not have significantly lower risk-adjusted in-patient or two-year mortality rates. CONCLUSIONS: Primary angioplasty is a highly effective option for AMI. PMID- 11028471 TI - Angiographic no-reflow phenomenon as a predictor of adverse long-term outcome in patients treated with percutaneous transluminal coronary angioplasty for first acute myocardial infarction. AB - OBJECTIVES: We sought to elucidate the long-term prognostic importance of angiographic no-reflow phenomenon after percutaneous transluminal coronary angioplasty (PTCA) for acute myocardial infarction (AMI). BACKGROUND: Angiographic no-reflow phenomenon, a reduced coronary antegrade flow (Thrombolysis in Myocardial Infarction [TIMI] flow grade < or =2) without mechanical obstruction after recanalization, predicts poor left ventricular (LV) functional recovery and survival in the early phase of AMI. We hypothesized that angiographic no-reflow phenomenon also predicts long-term clinical outcome. METHODS: We studied 120 consecutive patients with their first AMI treated by PTCA without flow-restricting lesions. The patients were classified as either no reflow (n = 30) or reflow (TIMI-3) (n = 90) based on post-PTCA cineangiograms to follow up (5.8 +/- 1.2 years) for cardiac death and nonfatal events. RESULTS: Patients with no-reflow had congestive heart failure (p < 0.0001), malignant arrhythmia (p = 0.038), and cardiac death (p = 0.002) more often than did those with reflow. Kaplan-Meier curves showed lower cardiac survival and cardiac event free survival (p < 0.0001) in patients with no-reflow than in those with reflow. Multivariate analyses disclosed that no-reflow phenomenon was an independent predictor of long-term cardiac death (relative risk [RR] 5.25, 95% confidence interval [CI] 1.85 to 14.9, p = 0.002) and cardiac events (RR 3.71, 95% CI 1.79 to 7.69, p = 0.0004). At follow-up, survivors with no-reflow had higher end diastolic and end-systolic LV volume indices and plasma brain natriuretic peptide levels, and lower LV ejection fractions (p = 0.0002, p < 0.0001, p = 0.002, p < 0.0001, respectively) than did those with reflow, indicating that no-reflow may be involved in LV remodeling. CONCLUSIONS: Angiographic no-reflow phenomenon strongly predicts long-term cardiac complications after AMI; these complications are possibly associated with LV remodeling. PMID- 11028472 TI - Evidence of prolonged inflammation in unstable angina and non-Q wave myocardial infarction. AB - OBJECTIVES: This study was designed to document the inflammatory response up to one year after acute presentation with unstable angina (UA) and non-Q wave infarction (NQMI) as reflected by the expression of soluble cell adhesion molecules (CAMs). BACKGROUND: Coronary plaque inflammation is a key component in the pathogenesis of acute coronary syndromes. Cell adhesion molecules are critical mediators of the inflammatory process. Soluble forms of these molecules are detectable in serum and are elevated acutely in patients with UA and NQMI. METHODS: Patients presenting with UA and NQMI had serum samples taken at presentation and then after three, six and 12 months. A control group of similar age and gender distribution was used for comparison. Levels of soluble inter cellular adhesion molecule-1, vascular cell adhesion molecule-1, endothelial selectin and platelet-selectin were measured using an ELISA technique. RESULTS: We studied 91 patients (M/F = 73/18, mean age 62 +/- 11 years, 56 UA and 35 NQMI) and 24 controls (M/F = 18/6, mean age 56 +/- 12 years). Levels of all four soluble CAMs were significantly elevated in both UA and NQMI patients at presentation, three and six months in comparison with controls. Levels in UA and NQMI groups fell between six and 12 months after initial presentation. CONCLUSIONS: The results suggest that the inflammatory stimulus triggering expression of CAMs is sustained for up to six months after presentation with either UA or NQMI and then returns toward control values over the following six months. PMID- 11028473 TI - Elevated homocysteine levels are associated with increased ischemic myocardial injury in acute coronary syndromes. AB - OBJECTIVES: This study was conducted to determine whether the amount of myocardial damage during acute coronary syndromes (ACS) is related to the admission plasma homocysteine concentration. BACKGROUND: Elevated homocysteine levels are associated with increased thrombosis in patients presenting with ACS. It is not known whether this association is reflected in the degree of myocardial injury in those patients. METHODS: We studied consecutive patients presenting with acute myocardial infarction (MI) (n = 205) and unstable angina pectoris (UAP) (n = 185). Plasma samples were collected on admission and prior to clinical intervention and were assayed for homocysteine by high performance liquid chromatography (HPLC). Myocardial necrosis was assessed by measurements of cardiac troponin T (cTnT) on admission and 12 h after admission (peak cTnT). The patients were studied by quintiles of homocysteine concentration. RESULTS: There was a significant increase in peak cTnT in the 5th homocysteine quintile in MI (analysis of variance [ANOVA], p = 0.005), the levels being 4.10, 3.86, 4.13, 6.20 and 7.85 microg/liter for quintiles 1 to 5, respectively (p < 0.0001, for top vs. bottom quintile). Similarly, there was a step-up in peak cTnT levels in the top homocysteine quintile in UAP (ANOVA, p < 0.0001), the levels being 0.03, 0.03, 0.02, 0.04 and 0.15 microg/liter, (p < 0.0001 for top vs. bottom quintile). In a multivariate regression model, the association between peak cTnT and the top homocysteine quintile remained strong after adjustment of other confounders including age, gender, final diagnosis and thrombolysis treatment (odds ratio [OR]: 2.92 (1.75-4.87) p < 0.0001). The patients with UAP were further examined according to peak cTnT levels below (cTnT negative) or above (cTnT positive) 0.1 microg/liter. Homocysteine levels were significantly higher in cTnT positive than cTnT negative patients; 13.8 (11.7-15.3) vs. 10.3 (9.4-11.3) micromol/liter, respectively, p = 0.002. CONCLUSIONS: Elevated homocysteine levels are associated with a higher risk of ischemic myocardial injury in patients presenting with ACS. PMID- 11028474 TI - Incidence and predictors of myocardial infarction among patients with atrial fibrillation. AB - OBJECTIVES: We sought to evaluate the utility of excluding myocardial infarction (MI) in patients presenting to the emergency department (ED) with atrial fibrillation (AF) and to identify predictors of MI in this group. BACKGROUND: Patients with AF are frequently admitted to the hospital, in part, to exclude an associated MI. There are no prospective data on unselected patients to support this common practice. METHODS: We conducted a prospective cohort study of all patients who presented to a single-center ED with the primary diagnosis of AF. RESULTS: Of a total of 255 patients, 190 (75%) were admitted to the hospital, and 109 of them underwent a standard "rule-out MI" protocol. Of these 109 patients, six (5.5%) were identified as having an acute MI at the time of admission. Chest pain was present in 39% of patients, with a sensitivity and specificity for the occurrence of MI of 100% and 65%, respectively. ST segment elevation or depression was present in 43% of patients, with a sensitivity and specificity of 100% and 51%. The presence of either major ST segment depression (>2 mm) or elevation on the admission electrocardiogram (ECG) was present in 6%, with a sensitivity of 100% and a specificity of 99%. The resulting positive and negative predictive values were 86% (95% confidence interval [CI] 42% to 99%) and 100% (95% CI 96% to 100%), respectively. Use of this criterion would have reduced the number of rule-out MIs in our study group by 94%, with no loss of sensitivity. CONCLUSIONS: Chest pain and ST segment depression are extremely common findings in patients presenting to the ED with AF and have limited power to predict MI. In contrast, ECG evidence of ST segment elevation or depression >2 mm appears to be a reliable discriminator of which patients are at risk for MI. Patients without significant ST segment changes are at very low risk for MI and may not require performance of the rule-out MI protocol or hospital admission if clinically stable. PMID- 11028475 TI - The impact of heavy passive smoking on arterial endothelial function in modernized Chinese. AB - OBJECTIVES: The study evaluated whether heavy exposure to environmental tobacco smoke (passive smoking) might damage arterial function in modernized Chinese. BACKGROUND: Heavy passive smoking is associated with arterial endothelial dysfunction in Caucasian, but not rural Chinese, subjects. METHODS: We studied 20 young (mean age 36.6 +/- 7.0 years) nonsmoking asymptomatic casino workers (9 men) in Macau who were exposed to environmental tobacco smoke for over 8 h/day for at least two years and 20 normal subjects (control subjects). These two groups were carefully matched for age, gender, body mass index (BMI), blood pressure, vessel diameter, cholesterol and glucose levels. Brachial artery diameter was measured by high-resolution B-mode ultrasound at rest, after flow increase (causing flow-mediated endothelium-dependent dilation) and after sublingual nitroglycerin (an endothelium-independent dilator). RESULTS: Flow mediated dilation (mean +/- SD% of diameter changes) was significantly lower in passive smokers (6.6 +/- 3.4%) compared with the controls (10.6 +/- 2.3%) (p < 0.0001). Nitroglycerin-induced dilation of the two groups were similar. Upon multivariate analysis, passive smoking exposure was the strongest independent predictor (beta = -0.59; p = 0.0001) for impaired flow-mediated endothelium dependent dilation (model R2 = 0.75, F value = 6.1, p = 0.0001). CONCLUSIONS: In modernized Chinese, as in Caucasians, exposure to heavy environmental tobacco smoke causes arterial endothelial dysfunction, a key early event in atherosclerosis. This may have serious implications for cardiovascular health in China, currently in a process of rapid modernization. PMID- 11028476 TI - Gender differences in sensitivity to adrenergic agonists of forearm resistance vasculature. AB - OBJECTIVES: The goal of this study was to investigate the mechanism of reduced vasoconstrictor sensitivity to norepinephrine in women compared with men. BACKGROUND: beta2-adrenergic agonists such as albuterol dilate forearm resistance vessels, partly by activating the L-arginine/nitric oxide pathway. Norepinephrine (which acts as beta- as well as alpha-adrenergic receptors) causes less forearm vasoconstriction in women than it does in men. This could be explained by a greater sensitivity to beta2-receptor stimulation in women than in men. METHODS: Forearm blood flow was measured by venous occlusion plethysmography in healthy women (days 10 to 14 of the menstrual cycle) and in men. Drugs were administered via the brachial artery in three separate protocols: albuterol +/- NG-monomethyl L-arginine (an inhibitor of nitric oxide synthase); substance P, nitroprusside and verapamil (control vasodilators); norepinephrine (+/- propranolol, a beta adrenergic receptor antagonist). RESULTS: Vasodilator responses to albuterol were greater in women than they were in men (p = 0.02 by analysis of variance). NG monomethyl-L-arginine reduced these similarly in men and women. Responses to control vasodilators were less in women than they were in men (each p < 0.05). Norepinephrine caused less vasoconstriction in women than it did in men (p = 0.02). Propranolol did not influence basal flow in either gender nor responses of men to norepinephrine but increased vasoconstriction to each dose of norepinephrine in women (p < 0.0001 for interaction between gender and propranolol). Responses to norepinephrine coinfused with propranolol were similar in men and women. CONCLUSIONS: Stimulation of beta2-adrenergic receptors causes greater forearm vasodilation in premenopausal women, at midmenstrual cycle, than it does in men. This is sufficient to explain why vasoconstriction to brachial artery norepinephrine is attenuated in such women. PMID- 11028477 TI - Basic fibroblast growth factor in patients with intermittent claudication: results of a phase I trial. AB - OBJECTIVES: This phase I study was designed to evaluate the safety, tolerability and pharmacokinetics of intra-arterial basic fibroblast growth factor (bFGF) in patients with atherosclerotic peripheral arterial disease (PVD) and intermittent claudication. We also assessed the effects of basic fibroblast growth factor (bFGF) on calf blood flow as a measure of biologic activity. BACKGROUND: Preclinical studies have shown that bFGF, an angiogenic peptide, promotes collateral development in animal models of myocardial and hind limb ischemia. The safety and efficacy of bFGF in patients is unknown, and early clinical trials are underway in coronary and peripheral arterial disease. METHODS: A double-blind, placebo-controlled, dose-escalation trial was conducted in patients with claudication demonstrating ankle/brachial index <0.8. Patients were randomly assigned to placebo (n = 6), 10 microg/kg of bFGF (n = 4), 30 microg/kg of bFGF once (n = 5) and 30 microg/kg of bFGF on two consecutive days (n = 4). Study drug was infused into the femoral artery of the ischemic leg. Detailed safety information including retinal photography for neovascularization were obtained through one year. Calf blood flow was measured with strain gauge plethysmography in the two higher dose treatment groups and in four placebo patients at baseline, one month and three to seven months after treatment. RESULTS: Intra-arterial bFGF was safe and well-tolerated. The half-life was 46 +/- 21 min. Calf blood flow increased at one month by 66 +/- 26% (mean +/- SEM) and at six months by 153 +/- 51% in bFGF-treated patients (n = 9, p = 0.002). Flow did not change significantly in the placebo group. CONCLUSIONS: In this initial randomized, double-blind, placebo-controlled trial in patients with atherosclerotic PVD and claudication, bFGF was well-tolerated. The data suggest a salutary biologic effect, and initiation of phase 2 trials is warranted. PMID- 11028478 TI - Therapeutic angiogenesis: time for the next phase. PMID- 11028479 TI - A common variant of the AMPD1 gene predicts improved cardiovascular survival in patients with coronary artery disease. AB - OBJECTIVE: We tested whether a common AMPD1 gene variant is associated with improved cardiovascular (CV) survival in patients with coronary artery disease (CAD). BACKGROUND: Reduced activity of adenosine monophosphate deaminase (AMPD) may increase production of adenosine, a cardioprotective agent. A common, nonsense, point variant of the AMPD1 gene (C34T) results in enzymatic inactivity and has been associated with prolonged survival in heart failure. METHODS: Blood was collected from 367 patients undergoing coronary angiography. Genotyping was done by polymerase chain reaction amplification and restriction enzyme digestion, resulting in allele-specific fragments. Coronary artery disease was defined as > or =70% stenosis of > or =1 coronary artery. Patients were followed prospectively for up to 4.8 years. Survival statistics compared hetero- (+/-) or homozygotic ( /-) carriers with noncarriers. RESULTS: Patients were 66 +/- 10 years old; 79% were men; 22.6% were heterozygous and 1.9% homozygous for the variant AMPD1(-) allele. During a mean of 3.5 +/- 1.0 years, 52 patients (14.2%) died, 37 (10.1%) of CV causes. Cardiovascular mortality was 4.4% (4/90) in AMPD1(-) allele carriers compared with 11.9% (33/277) in noncarriers (p = 0.046). In multiple variable regression analysis, only age (hazard ratio, 1.11/year, p < 0.001) and AMPD1(-) carriage (hazard ratio, 0.36, p = 0.053) were independent predictors of CV mortality. CONCLUSIONS: Carriage of a common variant of the AMPD1 gene was associated with improved CV survival in patients with angiographically documented CAD. The dysfunctional AMPD1(-) allele may lead to increased cardiac adenosine and increased cardioprotection during ischemic events. Adenosine monophosphate deaminase-1 genotyping should be further explored in CAD for prognostic, mechanistic and therapeutic insights. PMID- 11028480 TI - Prediction of coronary events with electron beam computed tomography. AB - OBJECTIVES: We sought to determine the prognostic accuracy of electron beam computed tomographic (EBCT) scanning of the coronary arteries at three to four years. BACKGROUND: Coronary artery calcium scores determined by EBCT correlate with the severity of coronary artery disease. However, previous reports of the prognostic accuracy of EBCT scanning for coronary events in asymptomatic individuals are conflicting. METHODS: Asymptomatic men and women undergoing coronary EBCT completed initial and follow-up evaluations, which included past medical history, the Rose angina questionnaire and interim cardiovascular events. Reported coronary events (death, nonfatal myocardial infarction [MI] and revascularization procedures) were confirmed without knowledge of the scan results. RESULTS: Information was obtained in 1,172 (99.6%) of 1,177 eligible subjects (baseline age 53 +/- 11 years, 71% men). During an average follow-up of 3.6 years, 39 subjects sustained coronary events: three coronary deaths, 15 nonfatal MIs and 21 coronary artery revascularization procedures. The mean coronary artery calcium score was 764 +/- 935 among subjects with events as compared with 135 +/- 432 among those without events (p < 0.0001). For the prediction of all coronary events and of nonfatal MIs and deaths, the areas under the receiver-operator characteristics curve were 0.84 and 0.86, respectively, and a coronary calcium score > or =160 was associated with odds ratios of 15.8 and 22.2, respectively. The odds ratios for all events remained high (14.3 to 20.2) after adjustment for self-reported cardiovascular risk factors. CONCLUSIONS: In asymptomatic adults, EBCT of the coronary arteries predicts coronary death and nonfatal MI and the need for revascularization procedures. PMID- 11028481 TI - The challenge of improving risk assessment in asymptomatic individuals: the additive prognostic value of electron beam tomography? PMID- 11028482 TI - Dobutamine-atropine stress echocardiography and dipyridamole sestamibi scintigraphy for the detection of coronary artery disease: limitations and concordance. AB - OBJECTIVES: We sought to compare dobutamine-atropine stress echocardiography (DASE) and dipyridamole Technetium 99-m (Tc-99m) sestamibi single photon emission computed tomography (SPECT) scintigraphy (DMIBI) for detecting coronary artery disease (CAD). BACKGROUND: Both DASE and DMIBI are effective for evaluating patients for CAD, but their concordance and limitations have not been directly compared. METHODS: To investigate these aims, patients underwent multistage DASE, DMIBI and coronary angiography within three months. Dobutamine-atropine stress echocardiography and stress-rest DMIBI were performed according to standard techniques and analyzed for their accuracy in predicting the extent of CAD. Segments were assigned to vascular territories according to standard models. Angiography was performed using the Judkin's technique. RESULTS: The 183 patients (mean age: 60 +/- 11 years, including 50 women) consisted of 64 patients with no coronary disease and 61 with single-, 40 with two- and 18 with three-vessel coronary disease. Dobutamine-atropine stress echocardiography and DMIBI were similarly sensitive (87%, 104/119 and 80%, 95/119, respectively) for the detection of CAD, but DASE was more specific (91%, 58/64 vs. 73%, 47/64, p < 0.01). Sensitivity was similar for the detection of CAD in patients with single vessel disease (84%, 51/61 vs. 74%, 45/61, respectively) and multivessel disease (91%, 53/58 vs. 86%, 50/58, respectively). Multiple wall motion abnormalities and perfusion defects were similarly sensitive for multivessel disease (72%, 42/58 vs. 66%, 38/53, respectively), but, again, DASE was more specific than DMIBI (95%, 119/125 vs. 76%, 95/125, respectively, p < 0.01). Dobutamine-atropine stress echocardiography and DMIBI were moderately concordant for the detection and extent of CAD (Kappa 0.47, p < 0.0001) but were only fairly (Kappa 0.35, p < 0.001) concordant for the type of abnormalities (normal, fixed, ischemia or mixed). CONCLUSIONS: Dobutamine-atropine stress echocardiography and DMIBI were comparable tests for the detection of CAD. Both were very sensitive for the detection of CAD and moderately sensitive for the extent of disease. The only advantage of DASE was greater specificity, especially for multivessel disease. Dobutamine-atropine stress echocardiography may be advantageous in patients with lower probabilities of CAD. PMID- 11028483 TI - Statistical methods to improve the precision of the treadmill exercise test. AB - OBJECTIVES: The study systematically compared different measures of ST segment depression from the treadmill exercise test. BACKGROUND: The value of the treadmill exercise test for objectively measuring treatment effects is limited by random error in the measurement of ST depression and may be biased by regression to the mean or by the decision to terminate the test. METHODS: Treadmill exercise was performed in 21 subjects with ischemic heart disease 1 h after isosorbide dinitrate 10 mg or placebo in a double-blind randomized crossover study. A 12 lead electrocardiogram (ECG) was recorded every 30 s during and at peak exercise. The relative sample size needed to detect the nitrate effect was compared for different summary measures of ST depression. RESULTS: The ST depression measured from a single unmatched lead at longest equivalent sub-maximal exercise needed the lowest sample size to detect the nitrate effect in paired comparisons (p = 0.000006). Averaging over multiple leads or times did not improve detection of the nitrate effect. The rate of increase in ST depression (in mm/min) calculated by linear regression needed a similar sample size (x1.32, 95% CI 0.62 to 2.58). A larger sample size was needed for ST depression at peak exercise (x2.9, CI 1.3, 11.1) and exercise duration (x4.5, CI 1.5, 38). Time to 1-mm ST depression was the least efficient measurement (relative sample size x15.5, CI 1.6, >1,000). Comparison of matched leads resulted in >2-fold differences in estimates of the nitrate effect because of bias from regression to the mean. CONCLUSIONS: Maximal ST depression at longest equivalent sub-maximal exercise and the maximal rate of increase in ST depression had less bias and random variation than did other commonly used measures. The rate of increase in ST depression is preferred because it can be calculated in either paired or unpaired studies. PMID- 11028484 TI - Modulation of beta1-adrenoceptor activity by domain-specific antibodies and heart failure-associated autoantibodies. AB - OBJECTIVES: Our study attempted to gain further understanding of the allosteric effects of human autoantibodies on beta1-adrenergic receptor (beta1-AR) function. BACKGROUND: Recently, we reported on the existence of activating anti-beta1-AR antibodies in patients with dilated cardiomyopathy (DCM 26% prevalence) or ischemic cardiomyopathy (ICM, 10% prevalence); however, their functional effects have not yet been thoroughly characterized. METHODS: In this study we detected functionally active receptor-antibodies in 8 out of 30 DCM patients. Their immunological and functional properties were analyzed using both synthetic receptor-peptides and intact recombinant human beta1-AR, and were compared with those of heterologous antibodies to selected beta1-AR domains generated in rabbits and mice. RESULTS: Rabbit, mouse, and human anti-beta1-AR against the second extracellular domain preferentially bound to a native receptor conformation and impaired radioligand binding to the receptor. However, their functional effects differed considerably: Rabbit and mouse antibodies decreased both basal and agonist-stimulated cAMP production, whereas the patient antibodies (n = 8) increased basal, and six of them also increased agonist-stimulated receptor activity (i.e., acted as receptor-sensitizing agents). Two out of eight human anti-beta1-AR increased basal but decreased agonist-stimulated receptor activity (i.e., acted as partial agonists). CONCLUSIONS: Antibodies against the same small beta1-AR domain can have very divergent allosteric effects, ranging from inhibitory to agonist-promoting activities. Activating autoantibodies were associated with severe cardiac dysfunction and thus might be involved in the development and/or course of human cardiomyopathy. PMID- 11028485 TI - Expression of tumor necrosis factor-alpha--converting enzyme and tumor necrosis factor-alpha in human myocarditis. AB - OBJECTIVES: We determined whether tumor necrosis factor-alpha-converting enzyme (TACE) is expressed with tumor necrosis factor-alpha (TNF-alpha) in myocarditis. BACKGROUND: Tumor necrosis factor-alpha-converting enzyme, which has recently been identified as belonging to the family of metalloproteinase disintegrin proteins, is responsible for the conversion of TNF-alpha precursor to its mature form. METHODS: We examined TACE and TNF-alpha expressions in endomyocardial biopsy tissues obtained from 14 patients with myocarditis and five control subjects by using quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry. RESULTS: Expression of TNF-alpha and TACE messenger ribonucleic acid (mRNA) was significantly greater in the myocarditis group than in the control group. A positive correlation was found between TNF-alpha and TACE mRNAs (r = 0.83, p < 0.05). Six patients with severe myocarditis underwent repeat biopsies. Although TNF-alpha and TACE mRNAs were expressed at high levels in the initial biopsies, a marked decrease was noted in the repeat biopsies. The immunostainings for TNF-alpha and TACE were positive in the myocytes and interstitial cells of myocardium obtained from patients with myocarditis. Expression of TACE and TNF-alpha mRNAs was greater in the subgroup in New York Heart Association functional class III or IV than in the subgroup in class I or II. Expression of TACE and TNF-alpha mRNA was correlated positively with left ventricular volume (TNF-alpha: r = 0.85; TACE: r = 0.80) and negatively with left ventricular systolic function (TNF-alpha: r = -0.85; TACE: r = -0.85). CONCLUSIONS: These findings indicate that the expression of TNF-alpha and TACE may have important implications in the pathogenesis of myocarditis and may influence advanced cardiac dysfunction in myocarditis. PMID- 11028486 TI - Prognostic value of pulmonary venous flow Doppler signal in left ventricular dysfunction: contribution of the difference in duration of pulmonary venous and mitral flow at atrial contraction. AB - OBJECTIVES: We assessed the contribution of difference in duration of pulmonary venous and mitral flow at atrial contraction (ARd-Ad) for prognostic stratification of patients with left ventricular (LV) systolic dysfunction. BACKGROUND: Although pulmonary venous flow (PVF) variables may supplement mitral flow patterns in evaluating left ventricular (LV) diastolic function, their value to the prognostic stratification of patients has not been investigated. METHODS: Pulsed wave Doppler mitral and PVF velocity curves were recorded in 145 patients (mean age: 70 years) with LV systolic dysfunction secondary to ischemic or nonischemic cardiomyopathy who were followed for 15 +/- 8 months. In 38% of patients, PVF signal was enhanced by the intravenous (IV) administration of a galactose-based echo-contrast agent. Based on E-wave deceleration time < or = or >130 ms and ARd-Ad, patients were grouped into restrictive (group 1, n = 40), nonrestrictive with ARd-Ad > or =30 ms (group 2, n = 55) and nonrestrictive with ARd-Ad <30 ms (group 3, n = 50). RESULTS: During follow-up, 29 patients died from cardiac causes and 28 were hospitalized for worsening heart failure (HF). On multivariate Cox model, ARd-Ad > or =30 ms provided important prognostic information with regard to cardiac mortality and emerged as the single best predictor of cardiac events (cardiac mortality, hospitalization). The 24-month cardiac event-free survival was best (86.3%) for group 3; it was intermediate (37.9%) for group 2; and it was worst (22.9%) for group 1 (p < 0.0002 group 1 vs. 3; p < 0.0005 group 2 vs. 3; p < 0.0003 group 1 vs. group 2). CONCLUSIONS: Assessment of ARd-Ad exhibited an independent value in the prognostic evaluation of patients with LV systolic dysfunction. Moreover, it contributed to identify patients at low, intermediate and high risk of cardiac events. PMID- 11028487 TI - The impairment of health-related quality of life in patients with intermittent atrial fibrillation: implications for the assessment of investigational therapy. AB - OBJECTIVES: We sought to assess the impact of intermittent atrial fibrillation (AF) on health-related quality of life (QoL). BACKGROUND: Intermittent AF is a common condition with little data on health-related QoL questionnaires to guide investigational therapies. METHODS: Outpatients from four centers, with documented AF (n = 152), completed validated QoL questionnaires (Medical Outcomes Study Short Form 36 [SF-36], Specific Activity, Symptom Checklist, Illness Intrusiveness and University of Toronto AF Severity Scales). Comparison groups were made up of healthy individuals (n = 47) and four cardiac control groups: published (n = 78) and created for study (n = 69) percutaneous transluminal coronary angioplasty (PTCA); published heart failure (n = 216) and published postmyocardial infarction (MI) (n = 107). RESULTS: Across all domains of the SF 36, AF patients reported substantially worse QoL than healthy controls (1.3 to 2.0 standard deviation units), with scores of 24%, 23%, 16% and 30% lower than healthy individuals on measures of physical and social functioning, mental and general health, respectively (all p < 0.001). Patients with AF were either significantly worse (p < 0.05, published controls) or as impaired (study controls) as either PTCA or post-MI patients on all domains of the SF-36 and the same as heart failure controls on SF-36 psychological subscales. Patients with AF were as impaired or worse than study PTCA controls on measures of illness intrusiveness, activity limitations and symptoms. Associations between objective disease indexes and subjective QoL measures had poor correlations and accounted for <6% of the total variability in QoL scores. CONCLUSIONS: Quality of life is as impaired in patients with intermittent AF as in patients with significant structural heart disease. Patients' perception of QoL is not dependent on the objective measures of disease severity that are usually employed. PMID- 11028488 TI - Marked ventricular repolarization abnormalities in highly trained athletes' electrocardiograms: clinical and prognostic implications. AB - OBJECTIVE: We sought to study the functional, clinical and prognostic implications of marked repolarization abnormalities (MRA) sometimes seen in athletes' electrocardiograms (ECGs). BACKGROUND: The clinical meaning of ECG MRA in athletes is unknown. No relationship has been drawn between either training intensity or any particular type of sport and MRA. Athletes are usually symptom free and do not show any decrease in their physical performance. It is as yet unclear whether MRA may have a negative effect on the performance of such athletes in competitive sports. METHODS: We studied 26 athletes with MRA (negative T waves > or =2 mm in three or more ECG leads at rest). No athletes presented clinical symptoms of cardiac disease or decrease in their physical performance. Clinical and physical examinations, ECG at rest, exercise test and echocardiographic and antimyosin studies were performed in all athletes. Rest/exercise myocardial perfusion single-photon emission computed tomography studies were performed in 17 athletes. The follow-up ranged from 4 to 20 years (mean 6.7 years). RESULTS: Four athletes were excluded due to hypertrophic cardiomyopathy. Echocardiographic studies showed right and left normal ventricular dimensions for highly conditioned athletes. In the exercise test, heart rate was 166 +/- 12.4 beats/min, and exercise tolerance was 15.2 +/- 2.7 metabolic equivalents of the task. All athletes had ECG at rest simulating myocardial ischemia or "pseudoischemia" with a tendency to normalize during exercise. Myocardial perfusion studies were normal in the studied athletes. Antimyosin studies showed mild and diffuse myocardial radiotracer uptake in 15 athletes (68%). No adverse clinical events were observed in the follow-up. CONCLUSIONS: These results suggest that MRA have no clinical or pathological implications in athletes and should, therefore, not preclude physical training or participation in sporting events. PMID- 11028489 TI - Glycoprotein IIIa Pl(A1/A2) polymorphism and sudden cardiac death. AB - OBJECTIVES: We studied the association of the Pl(A1/A2) polymorphism with coronary thrombosis, myocardial infarction (MI) and sudden cardiac death (SCD) in autopsied victims of sudden death. BACKGROUND: Sudden cardiac death is one of the leading symptoms of coronary heart disease in early middle age. Platelet glycoprotein (GP)IIb/IIIa fibrinogen receptors play a key role in coronary thrombosis and MI. Pl(A1/A2) polymorphism of the gene for GPIIIa has been previously studied in hospital MI patients. Significance of the Pl(A1/A2) polymorphism in victims of SCD is not known. METHODS: The Pl(A1/A2) polymorphism was studied in the Helsinki Sudden Death Study comprising 700 autopsied middle aged white Finnish men (33 to 70 years, mean 53 years) who suffered sudden or violent out-of-hospital death. RESULTS: Prevalence of the A2 allele decreased with age in the series. This decrease was observed among victims of SCD (n = 281) but not in men who died violently (n = 258) or of other diseases (n = 127). Of SCD victims below 50 years, 39.7% were carriers of the A2 allele compared with 28.3% among men under 50 who died of other causes (odds ratio [OR] 2.5, p = 0.01). Men with acute fatal coronary thrombosis (n = 39) were more often (OR 3.4, p < 0.01) carriers of the A2 allele than were men (n = 242) with SCD in the absence of acute coronary thrombosis (48.7% vs. 24.4%, respectively). In addition, men with MI and recent or old thrombosis (n = 67) were more often (OR 3.6, p = 0.005) carriers of the A2 allele than were men (n = 123) with MI in the absence of thrombosis (44.8% vs. 20.3%, respectively). These associations were especially strong in men under 60. CONCLUSIONS: Our results suggest that the A2 allele of the Pl(A1/A2) polymorphism of GPIIIa is a major risk factor of coronary thrombosis and may be one important predictor of SCD in early middle age. PMID- 11028490 TI - The ligament of Marshall: a structural analysis in human hearts with implications for atrial arrhythmias. AB - OBJECTIVES: We sought to study the anatomy of human ligament of Marshall (LOM). BACKGROUND: Although the LOM has been implicated in the genesis of focal atrial tachyarrhythmias, its gross anatomic and microscopic features in humans hearts have not been completely defined. METHODS: We studied seven postmortem human hearts from five men and two women with a mean age of 52 +/- 26 years. Four did not have any heart disease. One woman had dilated cardiomyopathy, and two men had chronic atrial fibrillation. A block of tissue encompassing the LOM from the coronary sinus (CS) cephalad, between the atrial appendage and left pulmonary veins, was dissected. Serial sections from this tissue were then stained with hematoxylin and eosin, trichrome, and/or tyrosine hydroxylase. RESULTS: The LOM consists of multiple sympathetic nerve fibers, ganglia, blood vessels and multiple myocardial tracts (Marshall Bundles) insulated by fibrofatty tissue. One or more myocardial tracts was inserted directly into the left atrial free wall and CS. The distance between insertion sites was 7.8 +/- 2.5 mm. Nerve fibers, some tyrosine hydroxylase positive, were present within the fibrofatty matrix and within the myocardial tracts. CONCLUSIONS: Human LOM 1) is innervated by sympathetic nerve fibers; 2) is more complex than the LOM in canine hearts; and 3) has multiple myocardial tract insertions into the left atrial free wall and CS, forming a substrate of reentry. Radiofrequency catheter ablation from the CS may fail to reach the free wall insertion. PMID- 11028491 TI - Cardiac troponin I predicts myocardial dysfunction in aneurysmal subarachnoid hemorrhage. AB - OBJECTIVES: We studied the incidence of myocardial injury in aneurysmal subarachnoid hemorrhage (SAH) using the more sensitive cardiac troponin I (cTnI) assay, correlated changes in cTnI with creatine kinase, MB fraction (CK-MB), myoglobin, and catecholamine metabolite assays, and examined the predictive value of changes in cTnI for myocardial dysfunction. BACKGROUND: Myocardial injury in aneurysmal SAH as evidenced by elevated CK-MB fraction has been reported. Little published data exist on the value of cTnI measurements in aneurysmal SAH. METHODS: Thirty-nine patients were studied for seven days. Clinical cardiovascular assessment, electrocardiographic (ECG), echocardiography, cTnI, CK, CK-MB and CK-MB index, myoglobin and 24-h urinary catecholamine assays were performed in all patients. The ECG abnormalities were defined by the presence of ST-T changes, prolonged QT intervals, and arrhythmias. An abnormal echocardiogram was defined by the presence of wall-motion abnormalities and a reduced ejection fraction. The severity of SAH was graded clinically and radiologically. RESULTS: Eight patients demonstrated elevations in cTnI (upper limit of normal is 0.1 microg/liter with the immunoenzymatic assay and 0.4 microg/liter with the sandwich immunoassay), while five had abnormal CK-MB levels (upper limit of normal is 8 microg/liter). Patients with more severe grades of SAH were more likely to develop a cTnI leak (p < 0.05). Patients with cTnI elevations were more likely to demonstrate ECG abnormalities (p < 0.01) and manifest clinical myocardial dysfunction (p < 0.01) as evidenced by the presence of a gallop rhythm on auscultation and clinical or radiological evidence of pulmonary edema as compared to those with CK-MB elevations. The sensitivity and specificity of cTnI to predict myocardial dysfunction were 100% and 91%, respectively, whereas the corresponding figures for CK-MB were 60% and 94%, respectively. Elevations in myoglobin levels (upper limit of normal <70 microg/liter) and urinary catecholamine metabolites (urinary vanilmandelate/creatinine ratio upper limit of normal, 2.6) are a nonspecific finding. CONCLUSIONS: Measurements of cTnI reveal a higher incidence of myocardial injury than predicted by CK-MB in aneurysmal SAH, and elevations of cTnI are associated with a higher incidence of myocardial dysfunction. Thus, cTnI is a highly sensitive and specific indicator of myocardial dysfunction in aneurysmal SAH. PMID- 11028492 TI - Short- and long-term results after thrombolytic treatment of deep venous thrombosis. AB - OBJECTIVES: The goal of this study was to assess the short- and long-term efficacy of different thrombolytic therapy regimens in patients with leg or pelvic deep venous thrombosis (DVT). BACKGROUND: It is unclear whether locoregional or systemic thrombolysis is superior in treating acute leg DVT or even whether lysis is more effective than anticoagulation therapy in preventing postthrombotic syndrome. METHODS: A total of 250 patients averaging 40 years of age with acute DVT were randomized into five groups to receive full heparinization (1,000 IU/h) and compression treatment, with four groups also administered locoregional tissue plasminogen activator (20 mg/day) or urokinase (100,000 IU/day) or systemic streptokinase (3,000,000 IU daily) or urokinase (5,000,000 IU daily). All groups then received anticoagulation and compression treatment for one year. Primary efficacy criteria included the change after one year in the number of closed vein segments and the occurrence of postthrombotic syndrome. RESULTS: Systemic thrombolytic therapy significantly reduced the number of closed vein segments after 12 months in patients with acute DVT compared with conventional treatment (p < 0.05). Postthrombotic syndrome also occurred with less frequency in systemically treated patients versus controls (p < 0.001). High dose thrombolysis led to better rates of complete recanalization after seven days (p < 0.01) than locoregional lysis. However, 12 patients receiving thrombolysis (9 systemic, 3 local) suffered major bleeding complications; 9 patients on systemic treatment developed pulmonary emboli. CONCLUSIONS: Systemic thrombolytic treatment for acute DVT achieved a significantly better short- and long-term clinical outcome than conventional heparin/anticoagulation therapy but at the expense of a serious increase in major bleeding and pulmonary emboli. Given the inherent risks for such serious complications, systemic thrombolysis, although effective, should be used selectively in limb-threatening thrombotic situations. PMID- 11028493 TI - Systolic anterior motion begins at low left ventricular outflow tract velocity in obstructive hypertrophic cardiomyopathy. AB - OBJECTIVES: The purpose of this study was to determine whether the dynamic cause for mitral systolic anterior motion (SAM) is a Venturi or a flow drag (pushing) mechanism. BACKGROUND: In obstructive hypertrophic cardiomyopathy (HCM), if SAM were caused by the Venturi mechanism, high flow velocity in the left ventricular outflow tract (LVOT) should be found at the time of SAM onset. However, if the velocity was found to be normal, this would support an alternative mechanism. METHODS: We studied with echocardiography 25 patients with obstructive HCM who had a mean outflow tract gradient of 82 +/- 6 mm Hg. We compared mitral valve M mode echocardiogram tracings with continuous wave (CW) and pulsed wave (PW) Doppler tracings recorded on the same study. A total of 98 M-mode, 159 CW, and 151 PW Doppler tracings were digitized and analyzed. For each patient we determined the LVOT CW velocity at the time of SAM onset. This was done by first determining the mean time interval from Q-wave to SAM onset from multiple M-mode tracings. Then, CW velocity in the outflow tract was measured at that same time interval following the Qwave. RESULTS: Systolic anterior motion began mean 71 +/- 5 ms after Q-wave onset. Mean CW Doppler velocity in the LVOT at SAM onset was 89 +/- 8 cm/s. In 68% of cases SAM began before onset of CW and PW Doppler LV ejection. CONCLUSIONS: Systolic anterior motion begins at normal LVOT velocity. At SAM onset, though Venturi forces are present in the outflow tract, their magnitude is much smaller than previously assumed; the Venturi mechanism cannot explain SAM. These velocity data, along with shape, orientation and temporal observations in patients, indicate that drag, the pushing force of flow, is the dominant hydrodynamic force that causes SAM. PMID- 11028495 TI - The fish mouth and three-dimensional echocardiography: new technology catches an old problem. PMID- 11028494 TI - Improved assessment of mitral valve stenosis by volumetric real-time three dimensional echocardiography. AB - OBJECTIVES: This study was performed to determine the feasibility, accuracy and reproducibility of real-time volumetric three-dimensional echocardiography (3-D echo) for the estimation of mitral valve area in patients with mitral valve stenosis. BACKGROUND: Planimetry of the mitral valve area (MVA) by two dimensional echocardiography (2-D echo) requires a favorable parasternal acoustic window and depends on operator skill. Transthoracic volumetric 3-D echo allows reconstruction of multiple 2-D planes in any desired orientation and is not limited to parasternal acquisition, and could thus enhance the accuracy and feasibility of calculating MVA. METHODS: In 48 patients with mitral stenosis (40 women; mean age 61 +/- 13 years) MVA was determined by planimetry using volumetric 3-D echo and compared with measurements obtained by 2-D echo and Doppler pressure half-time (PHT). All measurements were performed by two independent observers. Volumetric data were acquired from an apical view. RESULTS: Although 2-D echo allowed planimetry of the mitral valve in 43 of 48 patients (89%), calculation of the MVA was possible in all patients when 3-D echo was used. Mitral valve area by 3-D echo correlated well with MVA by 2-D echo (r = 0.93, mean difference, 0.09 +/- 0.14 cm2) and by PHT (r = 0.87, mean difference, 0.16 +/- 0.19 cm2). Interobserver variability was significantly less for 3-D echo than for 2-D echo (SD 0.08cm2 versus SD 0.23cm2, p < 0.001). Furthermore, it was much easier and faster to define the image plane with the smallest orifice area when 3-D echo was used. CONCLUSIONS: Transthoracic real-time volumetric 3-D echo provides accurate and highly reproducible measurements of mitral valve area and can easily be performed from an apical approach. PMID- 11028496 TI - Myocardial perfusion and function of the systemic right ventricle in patients after atrial switch procedure for complete transposition: long-term follow-up. AB - OBJECTIVES: Our purpose was to assess the right ventricular (RV) function and identify patients with RV impairment long after the Mustard or Senning operation. BACKGROUND: Systemic ventricular failure can cause myocardial perfusion abnormalities in thallium scintigraphy correlating with hemodynamic deterioration. METHODS: Myocardial perfusion at rest and at peak exercise was assessed in 61 patients, aged 7 to 23 years in mean time 10.0 +/- 2.9 years after surgery using technetium-99m methoxyisobutyl isonitrile single-photon emission computed tomography. Ventricular function was assessed by first-pass radionuclide angiography at rest. Exercise capacity was determined with a modified Bruce protocol. RESULTS: The mean RV ejection fraction was 36.1 +/- 7.7%, and left ventricular (LV) ejection fraction was 52.1 +/- 9.4%. Moderate or severe perfusion abnormalities on the rest scan were observed in 20 patients (33%). On exercise perfusion worsened in another 13 patients (21.3%). Patients with perfusion defects on stress scan had significantly lower RV and LV ejection fraction (33.2 vs. 39.4%; p = 0.002 and 49.2 vs. 55.5%; p = 0.01, respectively). They were also older (16.6 vs. 13.0 years; p = 0.002), operated on at an older age (4.0 vs. 2.4 years; p = 0.05) and had longer follow-up (12.5 vs. 10.5 years; p = 0.003). CONCLUSIONS: Myocardial perfusion defects are common findings in patients in long-term follow-up after atrial switch operation. Despite excellent exercise tolerance, the extent of myocardial perfusion abnormalities correlated well with impaired RV and LV function, and greater perfusion defects were seen more frequently in older patients with longer follow-up. It is likely that myocardial perfusion defects could be a sensitive predictor of systemic ventricular impairment. PMID- 11028497 TI - Coronary echocardiography in tetralogy of fallot: diagnostic accuracy, resource utilization and surgical implications over 13 years. AB - OBJECTIVES: This study sought to determine the diagnostic accuracy and impact of the systematic use of coronary echocardiography in a large group of preoperative patients with tetralogy of Fallot (TOF). BACKGROUND: Accurate preoperative identification of an anomalous coronary artery crossing the right ventricular outflow tract (RVOT) in patients with TOF is important to prevent coronary injury during surgical repair. METHODS: A retrospective review identified 598 patients with TOF between 1983 to 1995 who underwent an echocardiogram at <2 years old before complete surgical repair. Associated diagnoses included pulmonary stenosis (n = 433), pulmonary atresia (n = 121), common atrioventricular canal (n = 17), absent pulmonary valve syndrome (n = 24) and aortopulmonary window (n = 3). RESULTS: Based on intraoperative findings, 32 patients (5.4%) were found to have a major coronary artery crossing the RVOT. The use and diagnostic performance of coronary echocardiography increased over time, while the number of patients undergoing preoperative cardiac catheterization declined. During the most recent study period (1991 to 1995, n = 274), 97% of patients underwent coronary echocardiography yielding a sensitivity of 82%, specificity of 99% and accuracy of 98.5%. Of the 18 patients with TOF and pulmonary stenosis who had abnormal coronary arteries during this period, only 6 (33%) required an extracardiac conduit as part of their complete repair. CONCLUSIONS: Coronary echocardiography is an accurate noninvasive tool to delineate coronary anatomy in infants with TOF before complete repair. Routine preoperative cardiac catheterization solely for diagnosis of coronary anatomy is not necessary. The use of an extracardiac conduit can be avoided in the majority of patients with TOF and pulmonary stenosis who have a major coronary artery crossing the RVOT. PMID- 11028498 TI - Dichloroacetate improves postischemic function of hypertrophied rat hearts. AB - OBJECTIVES: We sought to determine whether improving coupling between glucose oxidation and glycolysis by stimulating glucose oxidation during reperfusion enhances postischemic recovery of hypertrophied hearts. BACKGROUND: Low rates of glucose oxidation and high glycolytic rates are associated with greater postischemic dysfunction of hypertrophied as compared with nonhypertrophied hearts. METHODS: Heart function, glycolysis and glucose oxidation were measured in isolated working control and hypertrophied rat hearts for 30 min before 20 min of global, no-flow ischemia and during 60 min of reperfusion. Selected control and hypertrophied hearts received 1.0 mmol/liter dichloroacetate (DCA), an activator of pyruvate dehydrogenase, at the time of reperfusion to stimulate glucose oxidation. RESULTS: In the absence of DCA, glycolysis was higher and glucose oxidation and recovery of function were lower in hypertrophied hearts than in control hearts during reperfusion. Dichloroacetate stimulated glucose oxidation during reperfusion approximately twofold in both groups, while significantly reducing glycolysis in hypertrophied hearts. It also improved function of both hypertrophied and control hearts. In the presence of DCA, recovery of function of hypertrophied hearts was comparable to or better than that of untreated control hearts. CONCLUSIONS: Dichloroacetate, given at the time of reperfusion, normalizes postischemic function of hypertrophied rat hearts and improves coupling between glucose oxidation and glycolysis by increasing glucose oxidation and decreasing glycolysis. These findings support the hypothesis that low glucose oxidation rates and high glycolytic rates contribute to the exaggerated postischemic dysfunction of hypertrophied hearts. PMID- 11028499 TI - Metabolism--the lost child of cardiology. PMID- 11028500 TI - Vascular repair after balloon overstretch injury in porcine model effects of intracoronary radiation. AB - OBJECTIVES: The purpose of this study was to evaluate the effect of IR on thrombus formation and dissection repair following overstretch balloon injury in porcine coronary arteries. BACKGROUND: Exposure of blood to the injured arterial wall after percutaneous transluminal coronary angioplasty (PTCA) induces thrombus formation and inflammation in the dissection plane. Neointima formation is related to smooth muscle cell (SMC) proliferation and migration into the preformed thrombus. Intracoronary radiation (IR) with doses of 10 to 25 Gy using either beta or gamma emitters can prevent neointima accumulation by reducing SMC proliferation. However, there are some indications that IR may delay the process of dissection repair after PTCA. The purpose of this study was to evaluate the effect of IR on thrombus formation and dissection repair after overstretch balloon injury in porcine coronary arteries. METHODS: Forty porcine coronaries were injured by balloon overstretch followed by either 0 or 18 Gy of 90Y prescribed to 1.2 mm from the balloon center. The animals were euthanized 14 days after treatment, and intimal area (IA) and IA corrected for medial fracture length (IA/FL) were quantified by digital image analysis. Dissections were quantified by tracing the length, thickness and area behind the dissection flap. The rate of dissections was calculated for each group. Thrombi were identified and designated as intraluminal thrombus or thrombus within dissection planes (mural thrombus), and area measurements were obtained. RESULTS: The irradiated group showed a significant reduction of IA/FL (0.55 +/- 0.29 vs. 0.05 +/- 0.09; p < 0.001). No difference was observed in the rate of dissection between control and irradiated arteries (77% vs. 88%, respectively). The control group showed a smaller dissection area (0.19 +/- 0.28 mm2 vs. 0.32 +/- 0.29 mm2; p < 0.05) with smaller mural thrombi (0.03 +/-0.01 mm2 vs. 0.29 +/- 0.30 mm2; p < 0.001). A strong correlation between dissection area and neointima area was observed only in the control group (R2 = 0.474; p < 0.003; alpha0.05 = 0.862). A positive correlation between mural thrombus and dissection area was observed only in the irradiated group (R2 = 0.889; p < 0.001; alpha0.05 = 1.00). CONCLUSIONS: These results suggest that the dissection area may be a useful parameter by which to quantify the extent of injury and repair after IR and may indicate an incomplete healing process after IR at this time point. PMID- 11028501 TI - Endoluminal reconstruction of the arterial wall with endothelial cell/glue matrix reduces restenosis in an atherosclerotic rabbit. AB - OBJECTIVES: The objectives of this study were 1) to improve the attachment of reimplanted endothelial cells (EC) using a fibrin glue, and 2) to assess the impact of endothelial reseeding on restenosis eight weeks after balloon angioplasty. BACKGROUND: A possible mechanism contributing to restenosis after balloon angioplasty is the loss of the EC lining. Previous attempts to reseed EC had little effect due to rapid loss of the seeded cells. METHODS: Twelve atherosclerotic rabbits were subjected to angioplasty of iliac arteries and reseeding procedure. One iliac artery was subjected to EC/glue reconstruction and a contralateral site to EC seeding without glue. The animals were sacrificed after 4 h. In another series 12 rabbits were treated in the same fashion and were restudied at eight weeks. Additionally, in 10 animals one iliac was subjected to glue treatment, and another served as control. RESULTS: Histological examination demonstrated the ability of this method to reattach the EC/glue matrix circumferentially to 68.0 +/- 6.7% of the arterial wall in comparison with 13.5 +/- 3.9% reattachment after EC seeding. Morphometry at eight weeks showed that the lumen area was significantly greater in the EC/glue group (1.23 +/- 0.35 mm2) than in the EC seeding alone (0.65 +/- 0.02 mm2) and 0.72 +/- 0.41 mm2 in the glue group. This was principally accounted for by the statistically significant differences in the intimal area (0.76 +/- 0.18 mm vs. 1.25 +/-0.26 mm2 and 1.01 +/- 0.53 mm2, respectively). CONCLUSIONS: The attachment of EC after angioplasty can be greatly improved with fibrin glue matrix. The near 70% endothelial coverage achieved by this method resulted in a significant reduction of restenosis in atherosclerotic rabbit. PMID- 11028502 TI - The class III antiarrhythmic effect of sotalol exerts a reverse use-dependent positive inotropic effect in the intact canine heart. AB - OBJECTIVES: We sought to study the rate related effects of sotalol on myocardial contractility and to test the hypothesis that the class III antiarrhythmic effect of sotalol has a reverse use-dependent positive inotropic effect in the intact heart. BACKGROUND: Antiarrhythmic drugs exert significant negative inotropic effects. Sotalol, a beta-adrenergic blocking agent with class III antiarrhythmic properties, may augment contractility by virtue of its ability to prolong the action potential duration (APD). METHODS: In 10 anesthetized dogs, measurements of left ventricle (LV) peak (+)dP/dt and simultaneous endocardial action potentials were made during baseline conditions and after sequential administration of esmolol and sotalol. In addition, electrical and mechanical restitution curves were constructed at a basic pacing cycle length of 600 ms by introducing a test pulse of altered cycle length ranging from 200 ms to 2,000 ms. RESULTS: In the steady state pacing experiments, sotalol prolonged the APD in a reverse use-dependent manner; such an effect was not seen with esmolol. At cycle lengths exceeding 400 ms, LV (+)dP/dt was significantly higher with sotalol than it was with esmolol. There was a direct relation between APD and LV (+)dP/dt with sotalol (r = 0.46, p < 0.001), but there was no significant relation between APD and LV (+)dP/dt with esmolol (r = 0.27, p = NS). Results in the single beat (restitution) studies were qualitatively similar to the steady state results; APD (at cycle length >400 ms) and LV (+)dP/dt (at cycle length >600 ms) were significantly higher with sotalol than they were with esmolol. CONCLUSIONS: The reverse use-dependent prolongation of APD by sotalol is associated with a positive inotropic effect. PMID- 11028504 TI - The aggressive low density lipoprotein lowering controversy. AB - Recent clinical trials have provided unequivocal evidence of major cardiovascular benefits from low density lipoprotein (LDL) lowering with statins. However, the three critical unresolved questions about aggressive LDL lowering are the shape of the curve relating cardiac events to LDL, the best surrogate measurement for assessing therapeutic efficacy and the best target for LDL therapy. The relation between cardiac events and LDL is curvilinear, both epidemiologically and during therapy. The benefit of lipid lowering diminishes progressively and becomes difficult to detect at lower LDL levels without a very large sample size. Assessment of the benefits of lipid lowering is further confounded by differences in the level of pretreatment LDL and by the non-LDL lowering effects of statins. Both epidemiologic studies and large randomized clinical trials have produced conflicting results concerning the best LDL target. Failure to reduce the event rate in patients with pretreatment LDL <125 mg (Cholesterol And Recurrent Events [CARE] trial) alerts us to the risk of extrapolating epidemiologic data to clinical practice, yet subset analysis of some clinical trials suggests the greatest benefit appears in those patients with the lowest on-treatment LDL levels (Scandinavian Simvastatin Survival Study [4S]). This controversy should be resolved in the next few years by several important on-going trials. In the face of seemingly contradictory data from current clinical trials, we can only speculate that very aggressive LDL lowering to <80 mg/dl could be accompanied by a modest therapeutic benefit beyond the current recommendations of the National Cholesterol Education Program. If any benefit is observed, it will have to be balanced against a small potential for increased adverse events. PMID- 11028505 TI - OTC or not OTC--that is the question. PMID- 11028503 TI - Endothelin-1 as a protective factor against beta-adrenergic agonist-induced apoptosis in cardiac myocytes. AB - OBJECTIVES: The purpose of this study was to investigate the regulation of beta adrenergic agonist-induced apoptosis by endothelin-1 (ET-1) in cardiac myocytes. BACKGROUND: Numerous hormonal factors including norepinephrine and ET-1 are activated in patients with heart failure. These factors may be involved in the positive and negative regulation of myocardial cell apoptosis observed in failing hearts. Recently, it has been shown that norepinephrine can induce myocardial cell apoptosis via a beta-adrenergic receptor-dependent pathway. METHODS: Primary cardiac myocytes were prepared from neonatal rats. These cells were stimulated with the beta-adrenergic agonist isoproterenol (ISO) in the presence or absence of ET-1. RESULTS: The administration of 10(-7) mol/liter of ET-1 completely blocked Iso-induced apoptosis. An endothelin type A receptor antagonist, FR139317, negated the inhibitory effect of ET-1 on apoptosis, while the endothelin type B receptor antagonist BQ788 did not show such a negation. Endothelin-1 also inhibited apoptosis induced by a membrane-permeable cAMP analogue (8-Br-cAMP), which bypassed Gi. The effect of ET-1 was neutralized by an MEK-1-specific inhibitor (PD098059), a phosphatidylinositol 3'-kinase inhibitor (wortmannin) and its downstream pp70 S6-kinase inhibitor, rapamycin. CONCLUSIONS: These findings suggest that ET-1 represents a protective factor against myocardial cell apoptosis in heart failure and that this effect is mediated mainly through endothelin type A receptor-dependent pathways involving multiple downstream signalings in cardiac myocytes. PMID- 11028506 TI - President's page: academic health centers: the making of a crisis and potential remedies. PMID- 11028507 TI - Alert to physicians: possible interaction of aggrenox and adenosine. PMID- 11028508 TI - Pharmacists agree: stop the pharmacy madness. PMID- 11028509 TI - The pharmacy perspective on managed care madness. PMID- 11028510 TI - Coronary anatomy in acute myocardial infarction patients with sudden out-of hospital death. PMID- 11028511 TI - Underestimation of the valvulopathy effect of fenfluramine. PMID- 11028512 TI - Risk of neutropenia with clopidogrel. PMID- 11028513 TI - Improvement of myocardial blood flow to ischemic regions by angiotensin converting enzyme inhibition. PMID- 11028514 TI - Pressure relaxation of the left ventricle and filling pressures. PMID- 11028515 TI - Calculation of resistance to pulmonary perfusion. PMID- 11028516 TI - American College of Cardiology/American Heart Association Clinical Competence statement on stress testing: a report of the American College of Cardiology/American Heart Association/American College of Physicians--American Society of Internal Medicine Task Force on Clinical Competence. PMID- 11028517 TI - Optical aberrations of the human cornea as a function of age. AB - We investigated how the optical aberrations associated with the anterior surface of the human cornea change with age in a normal population. Aberrations were computed for a central part of the cornea (4, 5, and 6 mm in diameter) from the elevation data provided by a videokeratographic system. Measurements were obtained in 59 normal healthy, near-emmetropic [spherical equivalent lower than 2 diopters (D)] subjects of three age ranges: younger (20-30 years old), middle aged (40-50 years old), and older (60-70 years old). The average corneal radius decreased with age and the cornea became more spherical. As a consequence, spherical aberration was significantly larger in the middle-aged and older corneas. Coma and other higher-order aberrations also were correlated with age. The root mean square of the wave aberration exhibited a linear positive correlation (P < 0.003) with age for the three ranges of pupil diameter. Despite a large intersubject variability, the average amount of aberration in the human cornea tends to increase moderately with age. However, this increase alone is not enough to explain the substantial reduction previously found in retinal image quality with age. The change in the aberrations of the lens with age and the possible loss of part of the balance between corneal and lenticular aberrations in youth may be the main factors responsible for the reduction of retinal image quality through the life span. PMID- 11028518 TI - Aliasing for rapidly counterphasing stimuli: a failure to demonstrate an M-cell sampling limit to resolution. AB - We investigated whether resolution is sampling limited for stimuli optimized for detection by magnocellular mechanisms. We measured peripheral (15 degrees and 30 degrees) spatial detection and resolution thresholds using 50% and 90% contrast flicker-defined gratings (25 Hz) and 90% contrast counterphasing sinusoidal gratings (25 Hz). Direction-discrimination performance for 90% contrast counterphasing sinusoidal gratings (25 Hz) was measured foveally. Our results indicate that resolution of rapidly counterphasing stimuli is sampling limited in peripheral vision but is consistent with limiting of performance by parvocellular mechanisms. Also, undersampling may not be necessary to account for motion reversals observed with gratings that both drift and flicker. PMID- 11028519 TI - Characterization of natural illuminants in forests and the use of digital video data to reconstruct illuminant spectra. AB - We describe illumination spectra in forests and show that they can be accurately recovered from recorded digital video images. Natural illuminant spectra of 238 samples measured in temperate forests were characterized by principal-component analysis. The spectra can be accurately approximated by the mean and the first two principal components. Compared with illumination under open skies, the loci of forest illuminants are displaced toward the green region in the chromaticity plots, and unlike open sky illumination they cannot be characterized by correlated color temperature. We show that it is possible to recover illuminant spectra accurately from digital video images by a linear least-squares-fit estimation technique. The use of digital video data in spectral analysis provides a promising new approach to the studies of the spatial and temporal variation of illumination in natural scenes and the understanding of color vision in natural environments. PMID- 11028520 TI - Quantitative analysis of error bounds in the recovery of depth from defocused images. AB - Depth from defocus involves estimating the relative blur between a pair of defocused images of a scene captured with different lens settings. When a priori information about the scene is available, it is possible to estimate the depth even from a single image. However, experimental studies indicate that the depth estimate improves with multiple observations. We provide a mathematical underpinning to this evidence by deriving and comparing the theoretical bounds for the error in the estimate of blur corresponding to the case of a single image and for a pair of defocused images. A new theorem is proposed that proves that the Cramer-Rao bound on the variance of the error in the estimate of blur decreases with an increase in the number of observations. The difference in the bounds turns out to be a function of the relative blurring between the observations. Hence one can indeed get better estimates of depth from multiple defocused images compared with those using only a single image, provided that these images are differently blurred. Results on synthetic as well as real data are given to further validate the claim. PMID- 11028521 TI - Noninterferometric quantitative phase imaging with soft x rays. AB - We demonstrate quantitative noninterferometric x-ray phase-amplitude measurement. We present results from two experimental geometries. The first geometry uses x rays diverging from a point source to produce high-resolution holograms of submicrometer-sized objects. The measured phase of the projected image agrees with the geometrically determined phase to within +/-7%. The second geometry uses a direct imaging microscope setup that allows the formation of a magnified image with a zone-plate lens. Here a direct measure of the object phase is made and agrees with that of the magnified object to better than +/-10%. In both cases the accuracy of the phase is limited by the pixel resolution. PMID- 11028522 TI - Theory of spatiochromatic image encoding and feature extraction. AB - We consider how to interpret, filter, and cross-correlate complex-value color (hue and saturation) images by using a single discrete Fourier transform: the spatiochromatic discrete Fourier transform. The model defines new types of spatiochromatic oriented sinusoidal gratings, termed rainbow gratings, which encode the variation of color over space. We demonstrate how color-opponent detectors observed within the vertebrate visual system can be easily defined by linear filters within this representation. This model also allows us to filter and detect both spatial and chromatic patterns in images by using a single cross correlation procedure. In doing so, we explore a new form of the Cauchy-Schwartz inequality applied to complex-valued scalar products. Results demonstrate the power of this form of spatiochromatic matched filtering in detecting signals embedded in such a significant amount of noise that they are not visible to the unaided human eye. PMID- 11028523 TI - Flatland optics: fundamentals. AB - "Flatland" is the title of a 120-year-old science fiction story. It describes the life of creatures living in a two-dimensional (2D) Flatland. A superior creature living in the three-dimensional (3D) spaceland, as we do, can easily inspect, for example, the inside of a Flatland house, as well as the content of a flat man's stomach without leaving any trace. Furthermore, the 3D person has supernatural powers that enable him to change the laws of physics in Flatland. We present here the concept of a 2D Flatland optics with one transversal coordinate x and one longitudinal coordinate z. The other transversal coordinate y allows total inspection of Flatland optics, and the freedom to change the wavelength, without using something like nonlinear optics or a Doppler shift. Monochromatic 3D light can be converted reversibly into polychromatic 2D light. A large variety of 2D systems and 2D effects will be presented here and in follow-up contributions. An epilogue faces the question, how "real" is Flatland optics? PMID- 11028524 TI - Calculation of diffraction effects on the average phase of an optical field. AB - We report on algorithms for the computation of the average phase of a beam over a detector in the near field. The basic idea is to reconstruct the optical field numerically and then use a quadrature algorithm to evaluate the quantity of interest. The various algorithms that employ discrete Fourier transform techniques for the computation of the field are described, and numerical tests that assess the accuracy of these algorithms are presented. No particular algorithm delivers the desired accuracy over the entire range of Fresnel numbers of interest, but each can produce satisfactory results within a particular range. Finally, new methods to evaluate the average phase are introduced, and their efficiency is assessed. PMID- 11028525 TI - Grating theory: new equations in Fourier space leading to fast converging results for TM polarization. AB - Using theorems of Fourier factorization, a recent paper [J. Opt. Soc. Am. A 13, 1870 (1996)] has shown that the truncated Fourier series of products of discontinuous functions that were used in the differential theory of gratings during the past 30 years are not converging everywhere in TM polarization. They turn out to be converging everywhere only at the limit of infinitely low modulated gratings. We derive new truncated equations and implement them numerically. The computed efficiencies turn out to converge about as fast as in the TE-polarization case with respect to the number of Fourier harmonics used to represent the field. The fast convergence is observed on both metallic and dielectric gratings with sinusoidal, triangular, and lamellar profiles as well as with cylindrical and rectangular rods, and examples are shown on gratings with 100% modulation. The new formulation opens a new wide range of applications of the method, concerning not only gratings used in TM polarization but also conical diffraction, crossed gratings, three-dimensional problems, nonperiodic objects, rough surfaces, photonic band gaps, nonlinear optics, etc. The formulation also concerns the TE polarization case for a grating ruled on a magnetic material as well as gratings ruled on anisotropic materials. The method developed is applicable to any theory that requires the Fourier analysis of continuous products of discontinuous periodic functions; we propose to call it the fast Fourier factorization method. PMID- 11028526 TI - Optical generation of focus wave modes. AB - Localized wave solutions of free-space wave equation can be used in numerous applications where the localized transmission of electromagnetic energy is of major importance. However, an optical implementation of localized wave fields has not been accomplished yet, except for an ultrashort version of the Bessel beams or the so called Bessel-X pulses. We propose an approach to constructing realizable optical schemes for generation of localized wave fields. We show that wavelength dispersion of the cone angle of axicons and circular diffraction gratings can be used to generate good approximation to focus wave modes. PMID- 11028527 TI - Plane waves in regular arrays of dipole scatterers and effective-medium modeling. AB - A simple analytical theory for finding eigensolutions for plane electromagnetic waves propagating along an axis in infinite regular arrays of small dipole particles is presented. The spacing between dipoles in every plane is assumed to be smaller than the wavelength; separation between the planes is arbitrary. The influence of evanescent modes is taken into account. This theory gives a model for an effective propagation constant that can be applied in a wide frequency range from the quasi-static regime to the Bragg reflection (photonic bandgap) region. PMID- 11028528 TI - Nonperturbative analysis of cross coupling in corrugated metal films. AB - There has been a recent renewal of interest in anomalously high energy transfer through otherwise opaque metal film geometries. One of the earliest such systems, that of surface plasmon cross coupling in a sinusoidally corrugated continuous metal film, was studied by Gruhlke et al. [Phys. Rev. Lett. 56, 2838 (1986)]. We show that it is possible to use a nonperturbative method to accurately predict this coupling, and we use this method to analyze both intrinsic coupling in symmetric structures and grating-assisted cross coupling in asymmetric structures. PMID- 11028529 TI - Analytical solution for the covariance and for the decorrelation time of the angle of arrival of a wave front corrugated by atmospheric turbulence. AB - The two-dimensional (2D) spatial covariance of the angle-of-arrival (AA) fluctuations is often used to investigate the properties of wave fronts corrugated by the atmosphere for high-angular-resolution techniques. Theoretical series expansions of this covariance are presented. The fast convergence of these series reduces the calculation time of the covariance done by numerical integration. The 2D covariance is a nonradial function. A physical interpretation of this anisotropy is proposed. The spatiotemporal correlation of the AA is deduced from the covariance assuming the "frozen-flow" hypothesis. The impact of the anisotropy on the evaluation of the number of predominant turbulent layers and on the corresponding winds is investigated, and an analysis of temporal correlations is performed. A simple theoretical approximation of the decorrelation time of the AA is given, which is found to be in agreement with experimental results. PMID- 11028530 TI - Isoplanatism in a multiconjugate adaptive optics system. AB - Turbulence correction in a large field of view by use of an adaptive optics imaging system with several deformable mirrors (DM's) conjugated to various heights is considered. The residual phase variance is computed for an optimized linear algorithm in which a correction of each turbulent layer is achieved by applying a combination of suitably smoothed and scaled input phase screens to all DM's. Finite turbulence outer scale and finite spatial resolution of the DM's are taken into account. A general expression for the isoplanatic angle thetaM of a system with M mirrors is derived in the limiting case of infinitely large apertures and Kolmogorov turbulence. Like Fried's isoplanatic angle theta0,thetaM is a function only of the turbulence vertical profile, is scalable with wavelength, and is independent of the telescope diameter. Use of angle thetaM permits the gain in the field of view due to the increased number of DM's to be quantified and their optimal conjugate heights to be found. Calculations with real turbulence profiles show that with three DM's a gain of 7-10x is possible, giving the typical and best isoplanatic field-of-view radii of 16 and 30 arcseconds, respectively, at lambda = 0.5 microm. It is shown that in the actual systems the isoplanatic field will be somewhat larger than thetaM owing to the combined effects of finite aperture diameter, finite outer scale, and optimized wave-front spatial filtering. However, this additional gain is not dramatic; it is less than 1.5x for large-aperture telescopes. PMID- 11028531 TI - Reconstruction and assessment of the least-squares and slope discrepancy components of the phase. AB - The concept of slope discrepancy developed in the mid-1980's to assess measurement noise in a wave-front sensor system is shown to have additional contributions that are due to fitting error and branch points. This understanding is facilitated by the development of a new formulation that employs Fourier techniques to decompose the measured gradient field (i.e., wave-front sensor measurements) into two components, one that is expressed as the gradient of a scalar potential and the other that is expressed as the curl of a vector potential. A key feature of the theory presented here is the fact that both components of the phase (one corresponding to each component of the gradient field) are easily reconstructable from the measured gradients. In addition, the scalar and vector potentials are both easily expressible in terms of the measured gradient field. The work concludes with a wave optics simulation example that illustrates the ease with which both components of the phase can be obtained. The results obtained illustrate that branch point effects are not significant until the Rytov number is greater than 0.2. In addition, the branch point contribution to the phase not only is reconstructed from the gradient data but is used to illustrate the significant performance improvement that results when this contribution is included in the correction applied by an adaptive optics system. PMID- 11028532 TI - Spectral analysis of vibrational harmonic motion by use of a continuous-wave CO2 Doppler lidar. AB - Vibrational motion of a harmonic oscillator was investigated with a focused continuous-wave (cw) CO2 Doppler lidar at 9.1-microm wavelength. A continuum of frequencies along with many discrete, equally spaced, resonant frequency modes was observed. The frequency modes are similar in structure to the oscillatory longitudinal modes of a laser cavity and arise because of interference of the natural resonant frequency of the oscillator with specific frequencies within the continuum. Each consecutive resonant frequency mode occurred for a movement of the oscillator much less than the wavelength of incident lidar radiation. For vigorous vibration of the oscillator, the observed spectra may be indicating nonlinear motion. PMID- 11028533 TI - Exterior caustics produced in scattering of a diagonally incident plane wave by a circular cylinder: semiclassical scattering theory analysis. AB - We use the semiclassical limit of electromagnetic wave scattering theory to determine the properties of the exterior caustics of a diagonally incident plane wave scattered by an infinitely long homogeneous dielectric circular cylinder in both the near zone and the far zone. The transmission caustic has an exterior/interior cusp transition as the tilt angle of the incident beam is increased, and each of the rainbow caustics has a farzone rainbow/exterior cusp transition and an exterior/interior cusp transition as the incident beam tilt angle is increased. We experimentally observe and analyze both transitions of the first-order rainbow. We also compare the predictions of the semiclassical approximation with those of ray theory and exact electromagnetic wave scattering theory. PMID- 11028534 TI - Family of detuning-insensitive phase-shifting algorithms. AB - The design of phase-shifting algorithms (PSA's) has been carried out with diverse strategies by different authors. A generalized algebraic approach is employed to obtain a family of detuning-insensitive PSA's; their behavior against a linear phase error is analyzed from a geometric point of view. The obtained results are compared with the conditions provided by the Fourier representation of the corresponding sampling reference functions. In our case, new equations as criteria for determining whether a PSA is detuning insensitive, new analytic expressions for the phase error, and new algorithms with interesting properties are achieved. PMID- 11028535 TI - General linear optical coordinate tranformations. AB - New optical configurations for performing general coordinate transformation operations of shear, rotation, and their combination are presented. These configurations consist of refractive spherical and cylindrical lenses that are readily available. Typically, high-resolution imagery can be obtained, depending on the size of the input object, the illumination wavelength, and the f-number of the lenses. Basic and more general configurations are presented, along with experimental results clearly showing image shearing, rotation, and a combination of these with high-quality output imagery. PMID- 11028536 TI - Resolution of a stochastic weakly damped nonlinear Schrodinger equation by a multilevel numerical method. AB - We consider a stochastic nonlinear Schrodinger equation related to signal propagation in waveguides and optical fibers. We first describe the modeling of the problem and the desired objectives concerning the transmission. We then present a new multilevel numerical method for its solution, which is based on a separation between low and high frequencies. We show that this method gives results of the same quality with significantly shorter CPU time compared with those of the other numerical methods commonly presented in the literature. PMID- 11028537 TI - Optical waveguide absorption sensor using a single coupling prism. AB - We propose an optical waveguide sensor that uses a leaky guided mode for measuring absorption of liquid samples. The sensor is composed of a single coupling prism on which a cladding layer and a waveguide layer are deposited. The guided mode generates dips in the reflectance spectrum; the depths of the dips depend on the extinction coefficient of a sample facing the layer. The sensitivity of the sensor is controlled by the thickness of the cladding layer. A simple theoretical model has been developed to analyze the behaviors of the sensor. In experiments we obtained sensitivity 17 times higher than that obtained by the conventional attenuated total reflection method. PMID- 11028538 TI - Providence: Surgical Infection Society 2000. PMID- 11028539 TI - Differential effect of caspase inhibition on proinflammatory cytokine release in septic patients. AB - The interleukins (IL)-1beta and IL-18 represent potent players in the proinflammatory cytokine cascade. Their activation is regulated predominantly through the IL-1-converting enzyme (ICE)/caspase-1. The role of caspases in the secretion of IL-1beta and IL-18, as well as in the release of the secondary induced cytokines IL-12 and interferon (IFN)-gamma in whole blood from septic patients compared to healthy controls, was studied. Inhibition of caspase activity by Z-VAD significantly reduced lipopolysaccharide (LPS) and Staphylococcus aureus (SAC) induced release of mature IL-1beta in septic patients and controls. In contrast, in whole blood from septic patients significantly elevated basal level of IL-18 were found, which could neither be further increased by LPS or SAC, nor be inhibited by Z-VAD. Release of IL-12 p40 was significantly lower in septic patients compared to controls and was not affected by Z-VAD. Despite high levels of IL-18, IFN-gamma was not detected in whole blood from septic patients even after stimulation with SAC or LPS. Thus, during sepsis, caspases participate in the processing of IL-1beta, whereas maturation of IL-18 during sepsis appears to be independent of caspases. The lack of IFN-gamma release seen in septic patients could be attributed to low IL-12 release rather than to diminished IL-18 release. PMID- 11028540 TI - Reduction of vancomycin-resistant enterococcal infections by limitation of broad spectrum cephalosporin use in a trauma and burn intensive care unit. AB - Both vancomycin and third-generation cephalosporin use are believed to contribute to a rise in vancomycin-resistant enterococci (VRE) infections. In 1998, the largest number of VRE infections in our hospital occurred in the trauma/burn intensive care unit (TBICU), accounting for nearly 20% of hospital infections. In an attempt to control the VRE infection rate, antibiotic protocols for prophylaxis, empiric, and definitive therapy were initiated during the final quarter of 1998 to minimize cephalosporin use by the introduction of piperacillin/tazobactam. Therefore, we undertook a study of the VRE infection rate for the TBICU in relation to vancomycin, piperacillin/tazobactam, piperacillin, third-generation cephalosporin, and total cephalosporin use before and after efforts to limit cephalosporins. These data were compared to those in the medical and surgical intensive care units. During 1998, seven VRE infections occurred in the TBICU. Following initiation of antibiotic protocols, one case of VRE infection occurred in the subsequent month and no cases in the 17 months since. The decrease in the VRE infection rate corresponded with a significant increase in the use of piperacillin/tazobactam and a reduction in third generation and total cephalosporin use. In contrast, cephalosporin use in the medical and surgical intensive care units remains significantly higher than in the TBICU, and neither unit has had a reduction in their VRE infection rates. PMID- 11028541 TI - Hypertonic saline inhibits neutrophil (PMN) priming via attenuation of p38 MAPK signaling. AB - Priming of the neutrophil cytotoxic response is central to the pathogenesis of early postinjury multiple organ failure (MOF). Platelet-activating factor (PAF) has been implicated as a key inflammatory mediator in postinjury neutrophil priming and requires p38 MAPK signaling to produce its biologic effects. Hypertonic saline (HTS) resuscitation decreases the postinjury inflammatory response following shock in animals and decreases receptor-mediated neutrophil (PMN) cytotoxic functions in vitro. We hypothesized that HTS attenuates PAF priming of the PMN cytotoxic response by interfering with PAF-mediated p38 MAPK signal transduction. Isolated PMNs were preincubated in isotonic buffer or HTS (Na+ = 180 mM), then primed with PAF. Neutrophil CD11b/CD18 expression was measured by flow cytometry. Receptor-dependent (fMLP), N-formyl-methionyl-leucyl phenylalanine, fMLP) and receptor-independent (PMA) O2- production was measured by reduction of cytochrome c in resting and PAF primed PMNs. Total p38 MAPK protein PAF-mediated p38 MAPK activation was assessed by western blot of PMN lysates. Clinically relevant levels of HTS attenuated PAF-mediated beta2-integrin expression. While HTS attenuated receptor-dependent (fMLP and PAF/fMLP) O2- production, receptor-independent (PMA) O2- production was unaffected. Conversely, HTS attenuated PAF priming of PMA-mediated O2- production. PAF and HTS did not alter total cellular p38 MAPK content. Clinically relevant levels of HTS alone did not activate p38 MAPK but inhibited PAF mediated p38 MAPK activation. HTS attenuates PAF priming of the PMN cytotoxic response by altering intracellular signal transduction. Therefore, HTS resuscitation may attenuate postinjury PMN priming and ultimately the risk of developing MOF. PMID- 11028542 TI - Impaired ex vivo lipopolysaccharide-stimulated whole blood tumor necrosis factor production may identify "septic" intensive care unit patients. AB - Currently, there is no reliable diagnostic test to identify septic intensive care unit (ICU) patients. We initiated studies to test the hypothesis that in sepsis, the in vivo exposure to endotoxin is detectable by the ex vivo analysis of lipopolysaccharide (LPS)-stimulated tumor necrosis factor (TNF) production. We obtained heparinized whole blood (WB) from 58 ICU patients and 14 healthy controls. The samples were incubated +/-10 ng/mL of LPS at 37 degrees C for 3 h. Plasma TNF levels were measured using enzyme-linked immunoassay (mean +/- standard error of the mean). Clinical data, including ICU length of stay (LOS), ventilator days (VentD), WBC, and positive cultures (Clt+), were obtained retrospectively. A wide range of LPS-stimulated WB TNF production (pg/mL) was observed in ICU patients (4481+/-469) and controls (6706+/-715). Patients were stratified into quartiles (I-IV) on the basis of the distribution of plotted LPS stimulated TNF values (pg/mL). Patients in quartile I (N = 14) had significantly lower TNF production (< 2000 pg/mL, P < 0.05) and required increased VentD (16 vs. 10 days, P < 0.05) compared to quartiles II-IV (N = 44). Patients in quartile I also had a higher incidence of infection (79 vs. 50%) and longer LOS (18 vs. 13 d) compared to quartiles II-IV. Impaired TNF release may be a manifestation of monocyte endotoxin tolerance and may be useful to diagnose sepsis. PMID- 11028543 TI - Phagocytosis of Candida albicans induces apoptosis of human neutrophils. AB - Neutrophil-mediated inflammation is terminated through the programmed cell death or apoptosis of the neutrophil, a process that can be inhibited by soluble mediators released during an inflammatory response. It has been reported, however, that the phagocytosis of intact bacteria can accelerate apoptosis. We evaluated the effects of the phagocytosis of a common nosocomial pathogen, Candida albicans, on the expression of apoptosis. Phagocytosis of killed Candida induced a dose-dependent increase in the apoptosis of normal neutrophils after 18 h of in vitro culture, from 40.7+/-9.1% to 81.7+/-4.5%, while supernatants from neutrophil:Candida co-cultures actually inhibited apoptosis. Induction of apoptosis was not dependent on phagocytosis, since opsonization of yeast with serum failed to increase apoptosis, while inhibition of phagocytosis with latrunculin B resulted in a slightly increased apoptotic rate. Increased apoptosis induced by Candida was associated with increased activity of the membrane-associated apoptotic enzyme, caspase 8, and with increased expression of the active form of the key executioner caspase, caspase 3. Increased apoptosis was associated with depletion of intracellular glutathione (GSH), and could be inhibited by the addition of exogenous GSH. These data demonstrate an important physiologic role for host-pathogen interactions in the resolution of inflammation and suggest that the response to an invading pathogen is an important stimulus to the restoration of normal immunologic homeostasis. PMID- 11028544 TI - TNFalpha-induced suppression of PMN apoptosis is mediated through interleukin-8 production. AB - Dysregulated neutrophil (polymorphonuclear PMN) apoptosis is thought to contribute to the onset of adult respiratory distress syndrome (ARDS) in critically ill patients. Tumor necrosis factor-alpha (TNFalpha), which is present in elevated levels in the bronchoalveolar lavage fluid in patients with ARDS, is thought to play a central role in regulating PMN function in the lungs. Studies have shown that short-term culture with TNFalpha increases apoptosis yet extended culture with TNFalpha suppresses apoptosis. However, it is unclear whether this latter effect of TNFalpha is directly or indirectly mediated through production of anti-apoptotic cytokines such as interleukin (IL)-8. To investigate the role of IL-8 in TNFalpha-induced apoptosis PMN were exposed to TNFalpha (100 ng/mL) in the presence or absence of antibodies to IL-8, and the extent of apoptosis was assessed. An enzyme-linked immunoassay was used to measure levels of the anti apoptotic cytokine IL-8, induced by TNFalpha-stimulation. Because TNFalpha may mediate its effect through various cell-signaling pathways, we next assessed the effect of kinase inhibition on the ability of TNFalpha to effect apoptosis and IL 8 production. Treatment with TNFalpha had a biphasic effect: at 4-8 h, apoptosis was increased but was markedly suppressed at 24 h (P < 0.05). PMN cultured for 24 h with TNFalpha also showed markedly increased levels of IL-8. Neutralization of IL-8 inhibited the ability of TNFalpha to suppress apoptosis (P < 0.05). Incubation of TNFalpha + p38-mitogen-activated protein kinase (MAPK) inhibitor SB202190 increased apoptosis (P < 0.01) and decreased IL-8 production to PMN control. To a lesser extent, incubation of TNFalpha with inhibitors to NF-kappaB (SN50) and PI3K (LY294002) also increased apoptosis and decreased IL-8 production (P < 0.05). These data illustrate a novel mechanism by which TNFalpha can indirectly elicit an anti-apoptotic effect via p38-MAPK induced release of the anti-apoptotic chemokine IL-8. The exploitation of such a pathway represents a potential target for regulation of PMN-mediated acute lung injury. PMID- 11028545 TI - Inhibited neutrophil apoptosis: proteasome dependent NF-kappaB translocation is required for TRAF-1 synthesis. AB - Neutrophil (PMN) apoptosis regulates local and systemic inflammation during sepsis. Tumor necrosis factor receptor-associated factors (TRAFs) have been implicated as mediators of apoptosis; however, the signaling pathways for their production in stimulated PMN are unclear. We hypothesize that NF-kappaB translocation is necessary for the induction of TRAF-1 in PMNs with prolonged survival. Neutrophils were isolated from the blood of healthy volunteers by Ficoll gradient centrifugation and red blood cell sedimentation. Neutrophil NF kappaB was inhibited with a proteasome inhibitor, PSI-I. Cells were treated with PSI-I (30 microM) or vehicle (DMSO 0.2%) for 50 min then incubated over an 18-h time course with LPS (10 to 1000 ng/mL), tumor necrosis factor alpha (TNFalpha) (2 to 20 ng/mL) or control media. In vitro apoptosis was quantified by propidium iodide FACS analysis. Total cellular TRAF-1 was detected by Western blot analysis of cell lysates. Steady state TRAF-1 mRNA was detected by RPA. NF-kappaB activity was determined by Western blot analysis for nuclear p65. Means and standard errors were calculated; data were analyzed by ANOVA. Lipopolysaccharide (LPS) and TNFalpha increased PMN nuclear p65 and steady state TRAF-1 mRNA. Apoptosis was inhibited by TNFalpha and LPS at 12 and 18 h (P < 0.01). Incubation of cells in the NF-kappaB inhibitor PSI-I blocked LPS and TNFalpha-induced inhibition of apoptosis (P < 0.05) and the induction of both nuclear p65 and TRAF-1 mRNA. These data demonstrate that inhibition of PMN apoptosis and TRAF-1 induction by LPS and TNFalpha is NF-kappaB dependent. PMID- 11028546 TI - Dopamine attenuates the chemoattractant effect of interleukin-8: a novel role in the systemic inflammatory response syndrome. AB - Activated neutrophil (PMN) adherence to vascular endothelium comprises a key step for both transendothelial migration and initiation of potentially deleterious release of PMN products. The biogenic amine, dopamine (DA), has been used for several decades in patients to maintain hemodynamic stability. The effect of dopamine on PMN transendothelial migration and adhesion receptor expression and on the endothelial molecules, E-selectin and ICAM-1, was evaluated. PMN were isolated from healthy controls, stimulated with lipopolysaccharide (LPS), and tumor necrosis factor-alpha (TNF-alpha) and treated with dopamine. CD 11b and CD 18 PMN adhesion receptor expression were assessed flow cytometrically. In a separate experiment, the chemoattractant peptide, IL-8, was placed in the lower chamber of transwells, and PMN migration was assessed. Human umbilical vein endothelial cells (HUVEC) were stimulated with LPS/TNF-alpha and incubated with dopamine. ICAM-1 and E-selectin endothelial molecule expression were assessed flow cytometrically. There was a significant increase in transendothelial migration in stimulated PMN compared with normal PMN (40 vs. 14%, P < 0.001). In addition, PMN CD11b/CD18 was significantly upregulated in stimulated PMN compared with normal PMN (252.4/352.4 vs. 76.7/139.4, P < 0.001) as were endothelial E selectin/ICAM-1 expression compared with normal EC (8.1/9 vs. 3.9/3.8, P < 0.05). After treatment with dopamine, PMN transmigration was significantly decreased compared with stimulated PMN (8% vs. 40%, P < 0.001). Furthermore, dopamine also attenuated PMN CD11b/CD18 and the endothelial molecules E-selectin and ICAM-1 compared with stimulated PMN/EC that were not treated dopamine (174/240 vs. 252/352, P < 0.05 and 4/4.4 vs. 8.1/9, P < 0.05. respectively). The chemoattractant effect of IL-8 was also attenuated. These results identify for the first time that dopamine attenuates the initial interaction between PMN and the endothelium, and consequently, modulates PMN exudation. Thus, biogenic amines, including dopamine, may function as anti-inflammatory cytokines. PMID- 11028547 TI - Effect of surfactant protein A (SP-A) on the production of cytokines by human pulmonary macrophages. AB - Surfactant protein A (SP-A) is thought to play a role in the modulation of lung inflammation during acute respiratory distress syndrome (ARDS). However, SP-A has been reported both to stimulate and to inhibit the proinflammatory activity of pulmonary macrophages (Mphi). Because of the interspecies differences and heterogeneity of Mphi subpopulations used may have influenced previous controversial results, in this study, we investigated the effect of human SP-A on the production of cytokines and other inflammatory mediators by two well-defined subpopulations of human pulmonary Mphi. Surfactant and both alveolar (aMphi) and interstitial (iMphi) macrophages were obtained from multiple organ donor lungs by bronchoalveolar lavage and enzymatic digestion. Donors with either recent history of tobacco smoking, more than 72 h on mechanical ventilation, or any radiological pulmonary infiltrate were discarded. SP-A was purified from isolated surfactant using sequential butanol and octyl glucoside extractions. After 24-h preculture, purified Mphi were cultured for 24 h in the presence or absence of LPS (10 microg/mL), SP-A (50 microg/mL), and combinations. Nitric oxide and carbon monoxide (CO) generation (pmol/microg protein), cell cGMP content (pmol/microg protein), and tumor necrosis factor alpha (TNFalpha), interleukin (IL)-1, and IL 6 release to the medium (pg/microg protein) were determined. SP-A inhibited the lipopolysaccharide (LPS)-induced TNFalpha response of both interstitial and alveolar human Mphi, as well as the IL-1 response in iMphi. The SP-A effect on TNFalpha production could be mediated by a suppression in the LPS-induced increase in intracellular cGMP. In iMphi but not in aMphi, SP-A also inhibited the LPS-induced IL-1 secretion and CO generation. These data lend further credit to a physiological function of SP-A in regulating alveolar host defense and inflammation by suggesting a fundamental role of this apoprotein in limiting excessive proinflammatory cytokine release in pulmonary Mphi during ARDS. PMID- 11028548 TI - Gender differences in sepsis: genetically determined? AB - In the pathogenesis of sepsis, tumor necrosis factor (TNF) release and host reaction may be genetically determined as demonstrated for TNFbeta Ncol polymorphism. Gender differences are considered as another important prognostic variable in patients with sepsis with better survival for women. The effect of sexual dimorphism on the genetic background of sepsis, however, is unknown. In a prospective study at two university hospital surgical intensive care units, (Bonn and Kiel), the role of the genomic marker TNFbeta Ncol polymorphism was evaluated with respect to gender. Two-hundred and one patients (68 women and 133 men) with severe sepsis were evaluated. A fragment of genomic DNA including the polymorphic site of the restriction enzyme Ncol was amplified by means of polymerase chain reaction. The genotype of each patient was determined after Ncol digestion of the amplified product. The genotype distribution of patients homozygous for TNFB1, heterozygous or homozygous for TNFB2 was comparable between men and women with severe sepsis. In women, no difference in survival rate was found between the different genotypes, while mortality rate was significantly increased in men homozygous for TNFB2 compared with the other genotypes (P < 0.05; P < 0.01, chi2 test). Overall, survival rate was higher for women (P < 0.05) but was not significantly different between men and women with respect to genotypes (P = 0.07 for TNFB2/B2). Poor prognosis of surgical sepsis can be determined by male gender and the genomic marker TNFbeta Ncol polymorphism which should be considered for further therapeutic interventions in sepsis. PMID- 11028549 TI - Biobrane improves wound healing in burned children without increased risk of infection. AB - A synthetic bilaminar membrane used as a skin substitute (Biobrane) has been shown to decrease pain and hospitalization in superficial second-degree burns. Despite these benefits, it has not been utilized universally, particularly in young children, due to a perceived increase in related infections. We propose that when this synthetic membrane is applied to superficial scald burns <25% of the total body surface area (TBSA), decreased healing times are expected without increased risk of infection. Between 1994-1999, 89 children treated within 48 h after receiving superficial partial thickness scald burns covering 5-25% TBSA with no indication of infection were seen at our hospital. Forty-one were assigned randomly to receive treatment with the skin substitute Biobrane and 48 to receive conservative treatment with topical antimicrobials and dressing changes. Comparisons of treatment were made between groups for length of hospitalization, wound healing times, and infectious complications. Children treated with Biobrane or topical antimicrobials were similar in age, race, sex, %TBSA burned, and location of burn. Those receiving Biobrane had shorter hospitalizations and healing times, which was significant for both infants and toddlers and older children. Treatment groups were not different in the use of systemic antibiotics or readmissions for infectious complications. Biobrane was removed in 5.9% of cases for non-adherence. The application of Biobrane within 48 h of superficial burns provides for shorter hospitalizations and faster healing times in children of all ages without increased risk of infection. PMID- 11028550 TI - Candida infection of local necrosis in severe acute pancreatitis is associated with increased mortality. AB - The aim of this study was to evaluate the frequency of Candida infection of pancreatic necrosis in patients suffering from severe acute pancreatitis (SAP) and to analyze its impact on the outcome. Two-hundred and fifty consecutive patients with SAP from January 1986 to December 1998 were studied retrospectively. Their mean APACHE II score at the day of admission was in 16.1 (range 8-35). All patients were in need of operative therapy. Overall mortality was 38.8% (97 patients). One-hundred and eighty-two patients (72.8%) suffered from local infected necrosis. Among these patients, local Candida infection was observed in 31 patients, whereof 23 patients (74%) suffered from local fungal infection detected at first operation. During the course of disease, 12 patients (39%) also revealed fungemia. Local Candida infection as compared to no Candida infection was associated with an increased mortality rate (84% vs. 32%; P 0.0001). Multivariate logistic regression analysis identified APACHE II score (P < 0.0001), age of the patient (P < 0.003), extent of pancreatic necrosis (P < 0.002), and local bacterial (P < 0.04) and fungal infection (P < 0.004) as independent factors significantly contributing to mortality. SAP, requiring surgical treatment, is associated with high in-hospital mortality. Patients suffering from local Candida infection are at high risk of fatal outcome. PMID- 11028551 TI - Health care resources consumed to treat postoperative infections: cost saving by perioperative immunonutrition. AB - The objectives of the study were to calculate the costs of postoperative complications and to evaluate whether the use of perioperative enteral immunonutrition, may lead to a saving in health care resources consumed. The economic analysis was based on data from a randomized double-blind trial that include 206 cancer patients who received perioperatively either enteral immunonutrition (treatment group, n = 102) or a standard enteral diet (control group, n = 104). Estimates of costs were based on resource use for treatment of complications, which were valued according to the National List of Sanitary Costs of the Italian Ministry of Health and on the medical Diagnosis-Related-Group (DRG) reimbursement rates. Costs of nutrition were also calculated. Cost comparison and cost effectiveness analyses were then carried out. Intent-to-treat analysis showed that the total costs of 52 postoperative complications were 322,218 euros, with a consumption of the DRG reimbursement rate of 15.4%. The costs of nutrition were 35,437 euros in the treatment group versus 10,768 euros in the control group. The total costs (nutrition plus treating complications) amounted to 113,778 euros in the treatment group versus 254,450 euros in the control group. The mean total costs per patient were 1,115 euros in the treatment group versus 2,447 euros in the control group (P = 0.04). Effectiveness was 83.3% in the treatment group versus 68.3% in the control group (P = 0.009). Cost effectiveness analysis showed a net saving of 2,386 euros per complication-free patient in favor of the treatment group. In conclusion, the perioperative use of immunonutrition appears cost effective due to a substantial saving of resources used to treat postoperative complications. PMID- 11028552 TI - Alternative case definitions of ventilator-associated pneumonia identify different patients in a surgical intensive care unit. AB - Diagnostic criteria that define ventilator-associated pneumonia (VAP) remain controversial. The purpose of this study was to evaluate common definitions of VAP and determine their relationship to each other and clinical treatment. This study prospectively evaluated several diagnostic criteria that define VAP in a cohort of 255 consecutive SICU patients ventilated for < 48 h. Definitions evaluated include the CDC definitions, the Johanson definitions which do not rely on culture data, the Physician's Probable diagnosis which relies on positive quantitative cultures, and the antibiotic treatment group. Forty-four patients (17%) received antibiotic treatment for VAP. Depending on the definition evaluated, criteria were met for a diagnosis of VAP from as low as 4% of patients by the Johanson definition to as high as 48% of patients by the CDC definition. There was poor agreement among the definitions in their ability to select the same patient as having VAP. Besides duration of mechanical ventilation and tube feeding, which were risk factors that predicted meeting the criteria for all groups, risk factors predicting VAP varied among the definitions. This study demonstrates that in a surgical ICU, the candidate definitions of pneumonia evaluated show little agreement. The particular case definition chosen to diagnose VAP will determine the incidence rate of pneumonia, the time to onset of pneumonia, and the risk factors of the type of patient treated. PMID- 11028553 TI - Community-acquired infections in the geriatric trauma population. AB - The incidence of community-acquired infections (CAs) and their relationship to the incidence of nosocomial infections (NI), to our knowledge, is unknown in elderly trauma patients. We prospectively collected data on 380 patients > or =65 years of age who were admitted >48 h to our trauma center over a 2-year period. One hundred seventy-seven patients (47%) developed an infection. A total of 147 (39%) patients were diagnosed with an NI, and 67 (18%) were diagnosed with a CA. Of the 67 patients with CA, 37 (55%) went on to develop an NI. Patients with the combination of CA and NI had the greatest mean ICU (28.6 days) and hospital length of stay (38.2 days). Mortality was increased significantly in patients with the combination of CA and NI (27%). Respiratory and genitourinary infections were the most common CA. Patients with respiratory CAs accounted for the greatest proportion of NIs. Thus, community-acquired and nosocomial infections significantly increase morbidity and mortality in elderly patients post-injury. Patients who present with a CA are at increased risk of acquiring an NI, which is associated with the most significant increase in length of stay and mortality. PMID- 11028554 TI - Selective digestive tract decontamination and vancomycin-resistant enterococcus isolation in the surgical intensive care unit. AB - Vancomycin-resistant Enterococcus (VRE) has emerged as a significant nosocomial pathogen in the surgical intensive care unit (SICU). We wished to test the hypothesis that the use of selective digestive tract decontamination (SDD) in the SICU affects the frequency of VRE isolation. A retrospective review of hospital records and the SICU database was performed using patients admitted to the SICU service for three or more days from January 1, 1996 to December 31, 1999 at our large tertiary-care teaching hospital. During this time use of SDD in selected patient populations decreased due to physician preference. Information gathered included length of SICU stay, presence of VRE infection or colonization, and use and duration of SDD protocol, vancomycin, and ceftazidime. There were 110 newly diagnosed VRE cases in the SICU during this time period. During the same time period 54 patients received SDD. Eight patients who received SDD had positive VRE cultures and seven had the initial positive culture after receiving SDD. Overall, 9.1% of eligible SICU patients received SDD, 18.5% of patients in the SICU for over 3 days had VRE, 7.3% of VRE patients received SDD, and 13.0% of the SICU patients who received SDD subsequently developed VRE. SDD use was not associated with VRE in univariate analysis. Logistic regression analysis showed higher odds ratios for SDD use in combination with vancomycin than for vancomycin use alone (OR=4.3 vs. 10.9). Odds ratios were over three times higher for SDD plus vancomycin plus ceftazidime use when compared to vancomycin plus ceftazidime use alone (OR=70.5 vs. 19.8). We conclude that administration of SDD alone did not correlate with increased VRE isolation, but that SDD use in conjunction with vancomycin and ceftazidime was associated with VRE isolation. PMID- 11028555 TI - The aromatase inhibitor, 4-hydroxyandrostenedione, restores immune responses following trauma-hemorrhage in males and decreases mortality from subsequent sepsis. AB - Studies have shown that immune responses are depressed in male mice, but not in proestrus females after trauma-hemorrhage (TH), resulting in increased mortality from subsequent sepsis in male mice compared with female mice. These gender specific alterations in immune function are believed to be due to differences in sex steroid levels. Aromatase is a key enzyme in the sex steroid biosynthesis. Although earlier studies have shown that aromatase inhibitors prevent thymic atrophy in aged male rats, it remains unknown whether the use of 4-hydroxy androstenedione (4-OHA) after TH in male mice has any salutary effects on the depressed immune responses. Male C3H/HeN mice were sham operated or subjected to trauma (i.e., midline laparotomy) and hemorrhagic shock (30+/-5 mmHg for 90 min) followed by adequate fluid resuscitation. 4-OHA (5 mg/kg) or vehicle was administrated s.c. just before resuscitation. At 2 h after resuscitation, the mice were killed, and spleens were harvested. Splenocyte proliferation, interleukin (IL-2), interferon (IFN-gamma), and IL-10 release and expression of androgen (AR) and estrogen receptors (ER)-alpha and -beta by immunoblot and reverse transcription-polymerase chain reaction (RT-PCR) were assessed. In another group, sepsis was induced by cecal ligation and puncture (CLP) 3 days after resuscitation, and survival was measured over a period of 10 days. A significant decrease in splenocyte proliferation, IL-2, and IFN-gamma release and increased release of IL-10 were observed in vehicle-treated mice. Animals treated with 4-OHA showed increased splenocyte proliferation, IL-2, and IFN-gamma release, and decreased IL-10 release. Immunoblot analysis showed decreased expression of the cytosolic AR, but no significant difference in the cytosolic and nuclear ER-alpha and -beta expression was observed in the vehicle-treated group after TH. In addition, AR and ER-beta mRNA expression was increased, whereas ER-alpha expression decreased in the vehicle-treated group after TH. ER alpha expression decreased and ER-beta expression increased in the nucleus of 4 OHA treated mice as determined by immunoblot. There was no difference in the cytosolic AR expression in the 4-OHA-treated group after TH. AR and ER-beta mRNA expression was unaffected, whereas ER-alpha expression increased under such conditions. In additional groups, the increased mortality rate after TH and subsequent sepsis was significantly reduced by 4-OHA treatment. Thus, 4-OHA seems to be a novel and useful adjunct for restoring the depressed immune functions in males after TH and for decreasing mortality rates from subsequent sepsis. PMID- 11028556 TI - Gender-related therapy: early IL-10 administration after hemorrhage restores immune function in males but not in females. AB - Recent experimental studies have found gender differences in the immune response after hemorrhagic shock with an enhanced immune function and lower mortality after subsequent sepsis in females than in males. Interleukin-10 (IL-10) has been shown to play a potential role in the treatment of early proinflammatory state after hemorrhagic shock. Although studies showed beneficial effects of the treatment with IL-10, it remains unclear whether the effects are gender related. To study this, male and female CBA/J mice were subjected to hemorrhage (35+/-5 mmHg for 90 min and fluid resuscitation) or sham operation. At resuscitation, each received either recombinant murine IL-10 (rmIL-10) or placebo i.p. At 48 h after resuscitation, peritoneal macrophages (pMphi) and splenocytes were harvested. IL-1beta and IL-12 release by pMphi and splenocyte proliferation and splenocyte IL-2 and interferon (IFN)-gamma release capacity were assessed. Interleukin-10 plasma levels were not increased after rmIL-10 treatment. The results indicate that rmIL-10 treatment restores depressed immune response (splenocyte proliferation, IFN-gamma, IL-1beta in males after hemorrhagic shock. In contrast, the immune responses after shock in females were not influenced by rmIL-10, with the exception of depressed splenocyte proliferation. In addition, sham-operated male mice treated with rmIL-10 showed immune depression compared with the placebo group. Thus, administration of rmIL-10 during resuscitation after hemorrhage produces salutary effects on the depressed immune responses in males but did not further enhance the immune functions in females under those conditions. PMID- 11028557 TI - Hepatocyte toll-like receptor 2 expression in vivo and in vitro: role of cytokines in induction of rat TLR2 gene expression by lipopolysaccharide. AB - We and others have demonstrated previously that cytokines, including interleukin 1 (IL-1) and tumor necrosis factor-alpha (TNFalpha), regulate LPS recognition proteins such as CD14 in the liver and on hepatocytes. Based on recent findings that the mammalian homologue of Drosophila Toll participates in LPS signaling, we examined the regulation of Toll-Like Receptor (TLR) gene expression by cytokines in vitro and its distribution in vivo with a focus on the liver as a site of host microbe interaction. Our results show that IL-1beta and/or TNFalpha participate in the upregulation of TLR2 mRNA levels in hepatocytes. Rats treated concurrently with LPS and antagonists of the IL-1 or TNFalpha receptor demonstrated significantly reduced LPS-induced hepatic expression of TLR2 compared to animals treated with LPS alone. The increase in hepatic TLR2 mRNA expression was associated with enhanced transcription as determined by nuclear run-on analysis. LPS treatment in vivo caused a marked TLR2 mRNA up-regulation in all of the tissues examined, with liver showing the highest expression. The high level of TLR2 expression in the liver may have important implications for pathogen-host interactions or microbial signaling. PMID- 11028558 TI - Inhibition of NF-kappaB by IkappaB prevents cytokine-induced NO production and promotes enterocyte apoptosis in vitro. AB - Nuclear factor-kappaB (N-kappaB) plays a key role in gut inflammation. NF-kappaB up-regulates proinflammatory genes encoding cytokines, adhesion molecules, and inducible nitric oxide synthase (iNOS). However, NF-kappaB has also been shown to up-regulate protective or anti-apoptotic factors. We utilized an adenoviral vector carrying a super-repressor form of the inhibitor of NF-kappaB, IkappaB, to examine the effects of NF-kappaB inhibition on cytokine-induced nitric oxide production and apoptosis in rat small intestinal epithelial cells (IEC-6). Chemical inhibitors of NF-kappaB, including pyrrolidine dithiocarbamate (PDTC), tosyl-lysine-chloromethylketone (TLCK), genistein, and N-acetyl-leu-leu norleucinal (n-LLnL) were also utilized. Treatment of AdIkappaB-transfected cells with cytomix [1000 U/mL IFN-gamma, 1 nM IL-1beta, and 10 ng/mL tumor necrosis factor alpha (TNFalpha)] or TNFalpha-containing cytokine combinations resulted in inhibition of cytokine-induced nitrite production and a marked increase in apoptosis compared to control cells. Apoptosis occurred independently of nitric oxide (NO) production since exogenous sources of NO did not inhibit apoptosis. Inducible NOS and clAP were down-regulated in AdIkappaB-transfected cells treated with cytomix. TLCK and LLnL treatment also induced apoptosis in cytomix-treated cells, while PDTC and genistein did not. Thus, although NF-kappaB up-regulates various pro-inflammatory genes, it may also have protective or anti-apoptotic effects in enterocytes. NF-kappaB appears necessary for upregulating cIAP in IEC 6 cells upon cytokine exposure. PMID- 11028560 TI - Activated neutrophils induce nitric oxide production in Kupffer cells. AB - Neutrophils (PMN) are proposed to contribute to hepatic dysfunction during sepsis. Transmigrating PMN have been demonstrated to adhere to and injure parenchymal cells (hepatocytes); however, the effect of sepsis-activated PMN on hepatic macrophages or Kupffer cells (KC) is poorly characterized. We hypothesize that PMN influence KC inflammatory mediator production, including nitric oxide. Rodent KC were co-cultured with PMN obtained from controls (Norm-PMN) or endotoxemic rats [lipopolysaccharide (LPS)-PMN] for 18 h. After an 18-h incubation, supernatants and cell lysates of the KC were analyzed for nitric oxide (NO) production. Co-cultures with LPS-PMN/KC demonstrated significantly increased production of nitrite and up-regulation of inducible nitric oxide synthase (iNOS) protein compared to KC alone or Norm-PMN/KC co-cultures. Immunohistochemistry revealed preferential iNOS protein staining in the cytoplasm of KC cultured with LPS-PMN compared to controls. Nitrite production in co cultures of KC and LPS-PMN where cell contact was inhibited by a cell impermeable but diffusable membrane was significantly reduced. These data provide evidence that KC can be stimulated directly by activated PMN for production of NO. Further, they suggest another mechanism by which PMN modulate hepatic function during sepsis. PMID- 11028559 TI - Blocking prostaglandin E2 after trauma attenuates pro-inflammatory cytokines and improves survival. AB - Major injury leads to impaired immune responses and increases the risk of infectious complications. Following trauma, increased prostaglandin E2 (PGE2) levels may be important in immunodysregulation. We hypothesized that blocking PGE2 with NS-398, a selective COX-2 inhibitor, during the first 24 h after injury may modify the immune response and protect the host from a subsequent septic challenge. BALB/c mice were given NS-398 (10 mg/kg) immediately after injury, at 12, and at 24 h after sham injury or trauma (femur fracture and 40% hemorrhage). On day 7 after injury, splenic macrophages were evaluated for cytokine production and COX-2 mRNA. In a separate study mice were injured, then given 3 doses of NS 398. After 7 days, cecal ligation and puncture was performed and mice were followed for survival. Traumatized mice given NS-398 had a significant survival advantage compared with trauma mice alone (P < 0.001). Macrophages from traumatized mice showed increased COX-2 mRNA and proinflammatory cytokines compared with controls (P < 0.05), whereas treatment of injured mice with NS-398 significantly decreased proinflammatory cytokine production (P < 0.05) and COX-2 mRNA. Therefore NS-398 given within 24 h of injury suppressed PGE2 through inhibition of cyclooxygenase, in addition to decreasing proinflammatory cytokines, and providing a survival advantage to the host. PMID- 11028561 TI - The transcription factor activator protein-1 is activated and interleukin-6 production is increased in interleukin-1beta-stimulated human enterocytes. AB - The intestinal mucosa is an active participant in the inflammatory and metabolic response to sepsis, endotoxemia, and other critical illness. The genes for various cytokines, e.g., interleukin 6 (IL-6), are regulated by multiple transcription factors, including nuclear factor-kappa B (NF-kappaB) and activator protein-1 (AP-1). In recent studies, treatment with IL-1beta of cultured Caco-2 cells, a human intestinal epithelial cell line, resulted in increased NF-kappaB DNA binding. The effect of IL-1beta on AP-1 activity in the enterocyte and the potential role of AP-1 in enterocyte IL-6 production are not known. We treated Caco-2 cells with IL-1beta and determined AP-1 activity by electrophoretic mobility shift assay (EMSA) and IL-6 production by enzyme-linked immunosorbent assay (ELISA). Treatment of Caco-2 cells with IL-1beta resulted in a dose- and time-dependent increase in AP-1 DNA binding. Supershift analysis suggests that activated AP-1 contained c-Jun, JunD, c-Fos, FosB, and Fra1 subunits. When Caco-2 cells were transiently transfected with an AP-1 luciferase reporter plasmid, stimulation with IL-1beta resulted in increased luciferase activity, suggesting that AP-1 DNA binding increased gene activation. Additional luciferase assays were performed with a plasmid containing a wild-type or AP-1-mutated IL-6 promoter. Stimulation of these cells with IL-1beta gave rise to results supporting the role of AP-1 in the regulation of IL-6 production. Geldanamycin, which has been shown in studies to inhibit AP-1 activation, blocked IL-1beta induced AP-1 luciferase gene activation and IL-6 production. These results suggest that the AP-1 family of transcription factors is activated by IL-1beta in human enterocytes and that AP-1 may at least in part regulate IL-6 production in these cells. PMID- 11028562 TI - The effect of burn injury on suppressors of cytokine signalling. AB - The newly identified suppressors of cytokine signaling (SOCS) family of proteins act as intracellular inhibitors of several cytokine signal transduction pathways. Their expression is induced by cytokine activation of the Janus kinase/signal transducer and activator of transcription (Jak/STAT) pathway, and they act as a negative feedback loop by subsequently inhibiting the Jak/STAT pathway either by direct interaction with activated Jaks or with the receptors. In this study we investigated the expression and translation of SOCS proteins after burn injury. Thermal injury increased the expression of SOCS3 compared with sham at 4 h, 24 h, and 10 days after thermal injury in the liver. SOCS3 protein was increased at 4 and 24 h after thermal injury in the liver. Expression of SOCS1 mRNA was not detected in sham or burn liver. SOCS2 mRNA and cytokine-inducible SH2-containing protein (CIS) mRNA were detected at the same levels for both sham and burn at all time points in the liver. In the spleen there was a trend towards an increase in SOCS1 mRNA at all time points; thermal injury significantly decreased SOCS2 mRNA compared with sham at 4 h, SOCS3 mRNA was significantly increased at 24 h compared with 10 days, and CIS mRNA was detected at the same levels for both sham and burn at all time points. In conclusion, thermal injury causes elevations in SOCS3 within 4 h after a burn, reaching a maximum at 24 h post injury. Levels continue to be elevated for up to 10 days post injury. SOCS3 may be very important in regulating the balance between immunosuppression and inflammation after thermal injury. PMID- 11028563 TI - Stat proteins play a role in tumor necrosis factor alpha gene expression. AB - Trauma produces dysfunction in immunity, which appears to be partially related to alterations in the cytokine response. Signal transducer and activator of transcription proteins (STATs) mediate activation of several cytokine genes. However, the effect of STAT proteins on tumor necrosis factor-alpha (TNFalpha) activation is not fully defined. We identified binding sites for STAT 3 and STAT 5/6 within the promoter region of TNFalpha and hypothesize that alterations in these sites would affect TNFalpha expression. The TNFalpha promoter was inserted into the luciferase reporter vector, and binding sites for STAT 3, STAT 5/6, and activator protein-1 (AP-1) were mutated using site-directed mutagenesis. Murine macrophages were transfected with the resultant plasmids, then incubated with and without lipopolysaccharide (LPS) or IFNalpha. Gene expression was measured by dual luciferase assay. Mutation of the STAT 3 binding site was associated with decreased LPS-inducible activity. Mutation of the AP-1 and STAT 5/6 consensus binding sites alone had no effect on TNFalpha expression. However, combined mutation of both STAT 5/6 and AP-1 was associated with increased LPS-inducible activity. Mutations of the STAT binding sites in the promoter region of TNFalpha affect TNFalpha gene expression. These results suggest a regulatory role for STATs in TNF gene transcription. PMID- 11028564 TI - The lipid fraction of post-hemorrhagic shock mesenteric lymph (PHSML) inhibits neutrophil apoptosis and enhances cytotoxic potential. AB - Dysfunctional neutrophil (PMN) apoptosis facilitates hyperinflammatory tissue injury. Previous work has demonstrated that post-hemorrhagic shock mesenteric lymph (PHSML) provokes PMN-mediated acute lung injury in animal models, but the mechanism remains unclear. We have documented that the lipid fraction of PHSML is responsible for PMN priming of the respiratory burst. In this study, we hypothesized that PHSML lipids delay PMN apoptosis and thereby further enhance PMN cytotoxic potential. Mesenteric lymph was collected from rats (n = 5) before (control), during non-lethal hemorrhagic shock (MAP 40 mmHg, 30 min), and during resuscitation (shed blood + 2x crystalloid). Human PMNs were incubated with control, PHSML, PHSML lipid extracts, and heat-treated PHSML (60 degrees C, 30 min.) at 1-10% (v:v) in RPMI 1640 for 24 h. Apoptosis was assessed using acridine orange/ethidium bromide staining and fluorescence microscopy. Priming of the respiratory burst was evaluated by incubating PMNs with (a) control PHSML or (b) PHSML lipid extracts for 24 h and by activating with fMLP (1 micromol/L). PHSML and PHSML lipid extracts (5-10%) inhibited PMN apoptosis. Heat denaturing the PHSML (to eliminate cytokines and complement) had no effect on the inhibition of PMN apoptosis. Similarly, incubation with polymixin B at a concentration that binds endotoxin had no effect. Both the PHSML and PHSML lipids (5%) following 24 h incubation primed the fMLP-activated oxidase. At physiologic concentrations, both PHSML and the lipid fraction of PHSML delay PMN apoptosis and prime the NADPH oxidase. These data further implicate the lipid components of mesenteric lymph as central in the pathogenesis of hemorrhagic shock induced PMN-mediated acute lung injury. PMID- 11028565 TI - In vivo and in vitro modulation of intercellular adhesion molecule (ICAM)-1 expression by hypertonicity. AB - Hepatic ischemia-reperfusion (I/R) is an important cause of organ dysfunction in the critically ill. With reperfusion, Kupffer cells release pro-inflammatory cytokines that promote endothelial cell (EC) expression of adhesion molecules such as intercellular adhesion molecule (ICAM)-1, facilitating neutrophil (PMN) infiltration. Studies suggest hypertonic saline (HTS) might exert beneficial effects on development of organ injury following shock on the basis of reduced PMN-EC interactions. We hypothesized that HTS alters expression of EC ICAM-1 and thus minimizes PMN-mediated injury. To test our hypothesis, we used an in vivo model of hepatic I/R and an in vitro model of activated EC. Rats underwent 30 min of hepatic ischemia after pretreatment with HTS (7.5% NaCl, 4cc/kg ia) or normal saline (NS). At 4 h reperfusion, plasma was taken for aspartate aminotransferase (AST) and liver tissue was harvested for assessment of hepatic ICAM-1 mRNA by Northern blot analysis. Human umbilical vein endothelial cells (HUVECs) were activated by lipopolysaccharide (LPS) and exposed to hypertonic medium (350-500 mOsM). HUVEC ICAM-1 protein was measured by cell ELISA and ICAM-1 mRNA by Northern blot analysis. HTS prevented hepatic I/R injury as measured by AST. AST of shams was 282.6+/-38.1 IU/L. I/R following NS pretreatment caused significant injury (AST 973.8+/-110.9 IU/L) compared to sham (SM) (P < 0.001). Pretreatment with HTS exerted significant protection following I/R with an AST of 450.9+/-56.3 IU/L (P < 0.05). There was no significant difference in AST levels between SM and HTS groups. Reduced hepatic injury after HTS and I/R was accompanied by inhibition of I/R-induced hepatic ICAM-1 mRNA expression compared to NS treated animals (P < 0.01). Similarly, hypertonicity inhibited HUVEC LPS-induced ICAM-1 protein (LPS: 1.86+/-0.19 absorbance units; 400 mOsM +/- LPS: 1.45+/-0.14 absorbance units; 450 mOsM + LPS: 1.02+/-0.19 absorbance units, P < 0.001) and mRNA expression. Thus, hypertonicity modulates endothelial ICAM-1 expression as one possible protective mechanism against I/R injury. PMID- 11028566 TI - Mesenteric lymph duct ligation provides long term protection against hemorrhagic shock-induced lung injury. AB - Recently we have shown that ligation of the main mesenteric lymph (MLN) duct prior to an episode of hemorrhagic shock (HS) prevents shock-induced lung injury. Yet, ligation or diversion of intestinal lymph immediately prior to injury is not clinically feasible. Diversion of intestinally derived lymph after injury to protect against secondary insults is possible, but it is not known how long the protective effects of lymph ligation would last. Thus, we tested whether ligation of the MLN duct seven days prior to HS would still be protective. Male Sprague Dawley rats were subjected to laparotomy with or without MLN duct ligation. Seven days later, half of the sham and actual MLN duct ligated animals randomly were selected to undergo HS (30 mmHG for 90 min). The other half of the animals was subjected to sham shock. Lung permeability, pulmonary myeloperoxidase (MPO) activity, and bronchoalveolar fluid (BALF) protein content were used to determine lung injury. Lymphatic division 7 days prior to HS continued to prevent shock induced lung injury as assessed by a lower Evans Blue dye concentration, BALF protein and MPO activity. In addition, there was no evidence of Patent Blue dye in the previously ligated MLN duct. Since ligation of the main mesenteric lymphatic duct continues to protect against shock-induced lung injury 1 week after duct ligation, it is feasible that lymphatic ligation performed after an injury remains protective against certain secondary insults for at least 1 week. PMID- 11028567 TI - Vulval pain syndromes. PMID- 11028568 TI - Physical health problems after childbirth and maternal depression at six to seven months postpartum. AB - OBJECTIVE: To investigate the relationship between maternal physical and emotional health problems six to nine months after childbirth. DESIGN: Statewide postal survey, incorporating the Edinburgh Postnatal Depression Scale, distributed to women six to seven months after childbirth, with telephone interview follow up of a sub-sample of participants at seven to nine months postpartum. PARTICIPANTS: The postal survey was distributed to all women who gave birth in a two-week period in Victoria, Australia in September 1993, except those who had a stillbirth or known neonatal death. Follow up interviews were conducted with respondents to the postal survey who provided contact details and expressed interest in participating in further research selected to recruit three groups of equivalent size according to scores on the Edinburgh Postnatal Depression Scale: a low scoring group (< 9); a borderline group (9-12); and a group with scores indicating probable depression (> or = 13). RESULTS: The response rate to the postal survey was 62.5% (n = 1,336). Respondents were representative of the total sample in terms of mode of delivery, parity and infant birthweight; young women, single women and women of non-English speaking background were under-represented. The participation rate in telephone follow up interviews was: 89.1% (n = 204), comprising 66 women with an Edinburgh Postnatal Depression Scale score of < 9; 72 women scoring 9-12; and 66 scoring > or =13. The point prevalence of depression at six to seven months postpartum was 16.9% (225/1,331, 95% CI 14.9-18.9%). Physical and related health problems associated with significantly increased odds of depression at six to seven months were: tiredness (OR 3.42 [2.2-5.3]); urinary incontinence (OR 2.23 [1.5-3.4]); back pain (OR 2.20 [1.6-3.0]); sexual problems (OR 2.16 [1.6-3.0]); more coughs, colds and minor illnesses than usual (OR 2.78 [1.9-4.1]); bowel problems (OR 1.93 [1.3-2.9]) and relationship difficulties (OR 3.88 [2.8-5.4]). At follow up, three physical health factors were associated with statistically significant linear trends with poorer levels of emotional wellbeing. These were: tiredness (chi2 for linear trend = 12.38, P < 0.001); urinary incontinence (chi2 for linear trend = 5.63, P = 0.02); and more minor illnesses than usual (chi2 for linear trend = 7.94, P = 0.005). CONCLUSION: The study confirms a link between maternal emotional wellbeing and physical health and recovery in the postnatal period and has important implications for clinical practice. Strategies for encouraging greater disclosure of physical and emotional health problems, assessment of the physical health of recent mothers presenting with depression, and emotional health of recent mothers presenting with other morbidity should be high priorities for all health professionals in contact with mothers in the year following childbirth. PMID- 11028569 TI - Women's health after childbirth: a longitudinal study in France and Italy. AB - OBJECTIVE: To describe the prevalence of various symptoms five months and twelve months after childbirth in two European countries, according to employment, financial difficulties and relationship with partner. DESIGN: Longitudinal multicentre survey with a similar design in France and Italy. Women were interviewed three times: at birth and twice after childbirth. At 12 months after childbirth, the survey was postal in both countries. POPULATION: Women who had been delivered of their first or second baby in three maternity units in France and five units in Italy. Women who had multiple pregnancies, a stillbirth or known neonatal death were excluded. RESULTS: The response rates were 83% in France and 88% in Italy. Fifteen symptoms were considered. The results were similar in the two countries and showed that the prevalence of most symptoms was higher at 12 months than at five months after childbirth. When their baby was one year old, more than half of the women reported backache, anxiety and extreme tiredness. Around one-third of women reported headache, lack of sexual desire, sleep disorders and depression. Piles, constipation and painful intercourse were also common. One year postpartum women with financial problems or a difficult relationship reported poor psychological health more frequently. CONCLUSION: Physical and emotional problems are common after birth, and they tend to increase over time. Backache, headache and piles can seriously interfere with day-to-day life. Sexual problems also may be a source of unhappiness for the woman and her partner. Extreme tiredness, anxiety and depression may make a woman feel guilty for not corresponding to the image of a healthy, happy and well-coping mother. There is a link between financial problems or a difficult relationship with her partner and her own wellbeing. Health professionals should be aware of the high prevalence of health problems among new mothers, and of the social context of women who are more likely to suffer from them. They should counsel the women in their care, in order to help them to find the best solution, be it medical or social in nature. PMID- 11028570 TI - Postpartum depression: identification of women at risk. AB - OBJECTIVE: To identify and test the predictive power of demographic, obstetric, and psychosocial risk factors of postpartum depression. DESIGN: Community-based, prospective follow up study based on questionnaires on past history of psychiatric disease, psychological distress and social support during pregnancy and depression at four months after delivery. Obstetric files were collected at time of birth. SETTING: Antenatal care clinic and delivery ward, Aarhus University Hospital, Denmark. POPULATION: 6,790 women giving birth between 1 January 1994 and 31 December 1995, who attended the antenatal clinic during pregnancy; 5,252 (78%) completed all questionnaires. The validation population comprised 528 women enrolled immediately prior to and after the study period. MAIN OUTCOME MEASURE: Postpartum depression four months after giving birth assessed by the Edinburgh Postnatal Depression Scale. RESULTS: 5.5% of the women suffered from postpartum depression, corresponding to a score of 13 or higher on the Edinburgh Postnatal Depression Scale. Risk factors identified by multivariate logistic regression analysis included psychological distress in late pregnancy (OR 6.3 [95% CI 4.4-9.1]), perceived social isolation during pregnancy (OR 3.6 [95% CI 1.9-7.0]); high parity (OR 3.8 [95% CI 1.8-8.0]); and a positive history of prepregnant psychiatric disease (OR 2.1 [95% CI 1.4-3.2]). No association was found between pregnancy or delivery complications, and postpartum depression. The maximum predictive power of the identified risk factors was 0.3. According to these results, one out of three women who suffers from psychological distress in late pregnancy with perceived social isolation will develop postpartum depression. CONCLUSION: Antenatal focus on psychosocial wellbeing may help to identify women at risk of postpartum depression. PMID- 11028572 TI - The effects of family planning education provided to different gender groups. AB - OBJECTIVE: To determine whether women or men should be preferred as the target group of the family planning education. DESIGN: Community trial (educational intervention study). SETTING: Three villages having similar characteristics in Gaziantep, Turkey. POPULATION: Currently married women aged between 15-49 and their husbands in these villages. METHODS: Family planning education was given only to women in the first village, only to men in the second village and to both women and their husbands in the third village. MAIN OUTCOME MEASURES: Changes in knowledge, attitudes, the behaviour of married women and their husbands between surveys performed before and after family planning education. RESULTS: Important changes in knowledge, attitudes, and behaviour were determined in all intervention groups after family planning education. The rate of effective contraceptive usage among women using any one of the contraceptive methods described significantly increased in all groups, especially where family planning education was given to both sexes. CONCLUSIONS: Family planning education given to one of the couples may easily affect the other. Consistent and regular education is more important than selecting target gender groups for family planning education. PMID- 11028571 TI - Levonorgestrel-releasing (20 microgram/day) intrauterine systems (Mirena) compared with other methods of reversible contraceptives. AB - OBJECTIVE: To assess the relative contraceptive effectiveness, tolerability and acceptability of the levonorgestrel-releasing (20 microg per day) intrauterine system (LNG-20) compared with reversible contraceptive methods in women of reproductive age. DESIGN: A systematic review and meta-analysis of randomised controlled trials. IDENTIFICATION: Studies were identified through seven databases, and by contacting investigators and organisations working in the contraceptive field. MAIN OUTCOME MEASURES: Unplanned pregnancy and continuation of contraceptive method. RESULTS: Five of the seven randomised controlled trials which met the inclusion criteria were included in the meta-analyses; four were comparisons of the LNG-20 intrauterine system with nonhormonal intrauterine devices. LNG-20 intrauterine systems were compared with intrauterine devices divided into two categories, those > 250 mm3 (Copper T 380 Ag and Copper T 380A intrauterine devices) and those < or = 250 mm3 (Nova-T, Copper T 220C and Copper 200 intrauterine devices). Pregnancy rates for the LNG-20 intrauterine system users were significantly less likely to become pregnant compared with users of intrauterine devices < or = 250 mm3, and significantly less likely to have an ectopic pregnancy. LNG-20 intrauterine system users were more likely to experience amenorrhoea and device expulsion than women using intrauterine devices > 250 mm3. LNG-20 intrauterine system users were significantly more likely than all the intrauterine device users to discontinue because of hormonal side effects and amenorrhoea. When the LNG-20 intrauterine system was compared with Norplant 2, the LNG-20 users were significantly more likely to experience oligo amenorrhoea, but significantly less likely to experience prolonged bleeding and spotting. CONCLUSIONS: The effectiveness of the LNG-20 intrauterine system was similar to or better than other contraceptive methods with which it was compared. Amenorrhoea was the main reason for the discontinuation of the LNG-20 intrauterine system, usually unnecessarily, since this end-organ suppression of bleeding is benign, associated with normal oestrogen levels. Women choosing this method should be informed of potential amenorrhoea when having pre-contraceptive counselling and that absent bleeding may be viewed as a positive outcome. PMID- 11028573 TI - Descriptive terms for women attending antenatal clinics: mother knows best? AB - OBJECTIVE: To determine the noun for 'women who attend antenatal clinics' that is most accepted by the women themselves. DESIGN: Cross sectional study. SETTING: Consultant-led antenatal clinics in Cornwall. POPULATION: All women attending consultant-led antenatal clinics over a two-month period. METHODS: The women were surveyed by written questionnaire. MAIN OUTCOME MEASURES: The first, second and third choices of descriptions offered to women attending antenatal clinics. Secondary outcome measures include the relation of maternal age, gestation, civil status, occupation and obstetric history to the individual's choice of description. RESULTS: Questionnaires were received from 446 women, constituting 13% of the antenatal population of Cornwall. Their median age was 28 years and median gestation 22 weeks; 255 (57%) had one or more children and 289 (65%) were married. The most popular choice of description was 'patient' (39% of first choices made), whereas the most accepted description was 'pregnant woman' (26% of totalled second and third choices). While women who selected 'patient' as first choice were slightly younger (mean 27.5 years) than the remaining women (mean 28.4 years), the choice of 'pregnant woman' was not related to any of the other recorded characteristics of the respondents. Commercial terms that consistently were selected least included 'client', 'consumer' and 'customer'. CONCLUSION: Some professional bodies and government organisations have criticised the use of the term 'patient' to describe antenatal women. In this, the largest study to investigate what the women themselves would choose, 'patient' is the most favoured term. PMID- 11028574 TI - The views and anticipated needs of women in early pregnancy. AB - OBJECTIVE: To assess the expectations of antenatal care of pregnant women at the outset of pregnancy. DESIGN: Questionnaire study within a randomised controlled trial, comparing traditional antenatal care with a more flexible schedule. SETTING: Eleven primary care centres providing midwifery care in Avon. POPULATION: Five hundred and ninety-three pregnant women at low risk of obstetric complications presenting for antenatal care. METHODS: A questionnaire was completed by women who agreed to participate in the trial shortly after antenatal booking. The questionnaire explored women's views on their attitudes to pregnancy and antenatal care, the locus of control related to pregnancy, the planning of the pregnancy and expectations of care. MAIN OUTCOME MEASURES: Comparisons between nulliparous and multiparous women in terms of their views of antenatal care, and their stated preference for a particular package of care. RESULTS: There was no difference in their views of pregnancy as an event entailing risk. On a locus of control scale that measured women's perceptions of factors which might affect their babies' health, nulliparous women rated antenatal care higher than multiparous women (P = 0.0001). However, this was not associated with any difference between the two groups in their stated preference for traditional or flexible care. Approximately half of the women expressed no preference, and of those who did 61% would opt for traditional care. Almost one-fifth of the whole sample welcomed the idea of flexible care. DISCUSSION: These data support the evidence of previous studies that there remains a strong desire among pregnant women to receive a 'traditional' pattern of care, even among those who have previously experienced normal pregnancy. However, a minority can be identified at the outset of pregnancy who may welcome a change to a more flexible pattern of care. PMID- 11028575 TI - A randomised controlled trial of flexibility in routine antenatal care. AB - OBJECTIVE: To assess changes in satisfaction associated with a flexible approach to antenatal care schedules offered to women at low obstetric risk. DESIGN: Randomised controlled trial. SETTING: Eleven primary care centres providing midwifery care in Avon. PARTICIPANTS: Six hundred and nine women at low risk of obstetric complications presenting for antenatal care. METHODS: A standard antenatal care schedule ('traditional care') was compared with a schedule based on a minimum number of visits and additional visits with timing agreed between women and midwives ('flexible care'). MAIN OUTCOME MEASURES: Women's attitudes to pregnancy and motherhood using a subscale of the Maternal Adjustment and Maternal Attitudes scale, satisfaction with antenatal care, and perception of the speed of recognition of antenatal complications. RESULTS: There was no difference between the two groups in terms of attitudes to pregnancy and motherhood (mean difference on Maternal Adjustment and Maternal Attitudes scale -0.64, 95% CI -1.39 to 0.11, P = 0.068) and no difference in the proportions of women reporting antenatal problems as soon as possible (traditional group 74.5%, flexible group 76.4%, difference -2%, 95% CI -12.1 to 8.2, P = 0.70). Women receiving traditional care reported higher levels of satisfaction for the care provided by community midwives (P < 0.01). Women receiving flexible care were more likely to report having a choice over the number and timing of their antenatal visits (P < 0.001), but were also more likely to report that they would like to have been seen more often (P < 0.01). There was no difference between the groups in rates of obstetric complications. CONCLUSIONS: An imposed reduction in antenatal visits has been reported to increase dissatisfaction in other studies. In this study, encouraging women to adopt a flexible approach to antenatal care resulted in a similar finding. Successful implementation of such approaches may depend on more careful selection of women who welcome such an approach, more encouragement to pregnant women to express their own needs and greater feelings of commitment on the part of the care providers. PMID- 11028576 TI - Subcutaneous versus intracutaneous injections of sterile water for labour analgesia: a comparison of perceived pain during administration. AB - OBJECTIVE: To investigate whether, during injections of sterile water, there is any difference in perceived pain between intracutaneous and subcutaneous injections. DESIGN: Blind controlled trial with cross-over design. SETTING: Goteborg and Skovde, Sweden. PARTICIPANTS: One hundred healthy female volunteers. METHODS: The women were randomised into two groups and subjected to two trials, within one week of each other. During the first trial one group (n = 50) received the intracutaneous injection first, followed by the subcutaneous injection. The second group (n = 50) was given the subcutaneous injection first, followed by intracutaneous injection. In both groups all the injections were given in reverse order during the second trial. MAIN OUTCOME MEASURES: Experienced pain during the administration of sterile water injections, measured by visual analogue scale. RESULTS: The analysis showed intracutaneous injections to be significantly more painful than subcutaneous injections, even after adjusting for injection day and for left/right site of injection (mean 60.8 vs 41.3, P < 0.001). CONCLUSIONS: The findings suggest that the less painful subcutaneous injection technique should be used. PMID- 11028577 TI - Expectant management of early onset, severe pre-eclampsia: maternal outcome. AB - OBJECTIVE: To evaluate the safety and outcome of women undergoing expectant management of early onset, severe pre-eclampsia. DESIGN: Prospective case series extending over a five-year period. SETTING: Tygerberg Hospital, a tertiary referral centre. POPULATION: All women (n = 340) presenting with early onset, severe pre-eclampsia, where both the mother and the fetus were otherwise stable. METHODS: Frequent clinical and biochemical monitoring of maternal status, together with careful blood pressure control, in a high care obstetric ward. MAIN OUTCOME MEASURES: Major maternal complications and prolongation of gestation. RESULTS: Multigravid women constituted 67% of the group. Antenatal biochemistry was reassuring with some expected, but not severe, deteriorations. Twenty-seven percent of women experienced a major complication, but few had poor outcomes. No maternal deaths occurred. Most major complications resolved quickly, necessitating only three admissions (0.8%) to the intensive care unit. One woman required dialysis. Pregnancies were prolonged by a mean (median) number of 11 days (9) before delivery, with more time being gained at earlier gestations. The postpartum inpatient stay (89% < or =7 days, bearing in mind that 82% of women were delivered by caesarean section) was not extended. CONCLUSION: Careful noninvasive management of early onset, severe pre-eclampsia in a tertiary centre can diminish and limit the impact of serious maternal complications. Valuable time to prolong the pregnancy and improve neonatal outcome is thereby gained. PMID- 11028578 TI - Expectant management of early onset, severe pre-eclampsia: perinatal outcome. AB - OBJECTIVE: To evaluate the perinatal outcome of expectant management of early onset, severe pre-eclampsia. DESIGN: Prospective case series extending over a five-year period. SETTING: Tertiary referral centre. POPULATION: All women (n = 340) presenting with early onset, severe pre-eclampsia, where both mother and the fetus were otherwise stable. METHODS: Frequent clinical and biochemical monitoring of maternal status with careful blood pressure control. Fetal surveillance included six-hourly heart rate monitoring, weekly Doppler and ultrasound evaluation of the fetus every two weeks. All examinations were carried out in a high care obstetric ward. MAIN OUTCOME MEASURES: Prolongation of gestation, perinatal mortality rate, neonatal survival and major complications. RESULTS: A mean of 11 days were gained by expectant management. The perinatal mortality rate was 24/1,000 (> or = 1,000 g/7 days) with a neonatal survival rate of 94%. Multivariate analysis showed only gestational age at delivery to be significantly associated with neonatal outcome. Chief contributors to neonatal mortality and morbidity were pulmonary complications and sepsis. Three pregnancies (0.8%) were terminated prior to viability and only two (0.5%) intrauterine deaths occurred, both due to placental abruption. Most women (81.5%) were delivered by caesarean section with fetal distress the most common reason for delivery. Neonatal intensive care was necessary in 40.7% of cases, with these babies staying a median of six days in intensive care. CONCLUSION: Expectant management of early onset, severe pre-eclampsia and careful neonatal care led to high perinatal and neonatal survival rates. It also allowed the judicious use of neonatal intensive care facilities. Neonatal sepsis remains a cause for concern. PMID- 11028579 TI - First trimester maternal serum free beta human chorionic gonadotrophin and pregnancy associated plasma protein A as predictors of pregnancy complications. AB - OBJECTIVE: To examine the value of first trimester maternal serum free beta human chorionic gonadotrophin (beta hCG) and pregnancy associated plasma protein A (PAPP-A) as predictors of pregnancy complications. DESIGN: Screening study. SETTING: Antenatal clinics. POPULATION: Singleton pregnancies at 10-14 weeks of gestation. METHODS: Maternal serum free beta hCG and PAPP-A were measured at 10 14 weeks of gestation in 5,584 singleton pregnancies. In the 5,297 (94.9%) pregnancies with complete follow up free beta hCG and PAPP-A were compared between those with normal outcome and those resulting in miscarriage, spontaneous preterm delivery, pregnancy induced hypertension or fetal growth restriction and in those with pre-existing or gestational diabetes. RESULTS: Maternal serum PAPP A increased and beta hCG decreased with gestation. The multiple of median maternal serum PAPP-A was significantly lower in those pregnancies resulting in miscarriage, pregnancy induced hypertension, growth restriction and in those with pre-existing or gestational diabetes mellitus, but not in those complicated by spontaneous preterm delivery. The level was < 10th centile of the reference range in about 20% of the pregnancies that subsequently resulted in miscarriage or developed pregnancy induced hypertension or growth restriction, and in 27% of those that developed gestational diabetes. Maternal serum free beta hCG was < 10th centile of the reference range in about 15% of the pregnancies that subsequently resulted in miscarriage or developed pregnancy induced hypertension or growth restriction, and in 20% of those that developed gestational diabetes. CONCLUSION: Low maternal serum PAPP-A or beta hCG at 10-14 weeks of gestation are associated with subsequent development of pregnancy complications. PMID- 11028580 TI - One stop clinic for assessment of risk for fetal anomalies: a report of the first year of prospective screening for chromosomal anomalies in the first trimester. AB - OBJECTIVE: To evaluate the introduction of a one stop multidisciplinary clinic for screening for fetal chromosomal abnormalities in the first trimester by a combination of maternal serum biochemistry and ultrasonography providing a risk of chromosomal abnormalities within a one hour clinic visit. DESIGN: One year retrospective review of screening performance. POPULATION: All women attending for routine antenatal care. The population included 4,190 singleton pregnancies in women of all ages screened between 10 weeks and 3 days and 13 weeks and 6 days of gestation between the periods 1 June 1998 and 31 May 1999 in a district general hospital antenatal clinic. METHODS: All women booked into the clinic were offered screening by a combination of maternal serum free beta human chorionic gonadotrophin (hCG) and pregnancy associated plasma protein A (PAPP-A) and fetal nuchal translucency thickness. Women at increased risk of carrying a fetus with trisomy 21 or trisomy 18/13 (> or =1 in 300 at sampling) were offered counselling and an invasive diagnostic procedure. Follow up of the outcome of all pregnancies was carried out. MAIN OUTCOME MEASURES: The detection rate for trisomy 21, trisomy 18/13 and all aneuploides, false positive rate, uptake of screening, uptake of chorionic villus sampling in women identified at increased risk and fetal loss after chorionic villus sampling. RESULTS: Overall 97.6% of the women (4,088/4,190) accepted first trimester screening. The rate of detection of trisomy 21 was 86% (6/7), for trisomy 18/13 100% (9/9) and for all aneuploides 95% (18/19). Fetal death at presentation was found in 1.6% of pregnancies (69/4,088). Of women who accepted screening, 6.1% (257/4,088) presented too late for fetal nuchal translucency measurement and 6.5% of the women (271/4,088) presented too early. The false positive rate was 6.7% (253/3,762). Uptake of invasive testing was 83% (207/253). CONCLUSION: First trimester prenatal screening for chromosomal abnormalities using a combination of maternal serum biochemistry and fetal nuchal translucency thickness can achieve detection rates in excess of 90%. These services can be provided in a one stop multidisciplinary clinic. PMID- 11028581 TI - Cervicovaginal fibronectin and cervical length at 23 weeks of gestation: relative risk of early preterm delivery. AB - OBJECTIVES: To establish the prevalence of cervicovaginal fetal fibronectin positivity at 23 weeks of gestation in a routine population of singleton pregnancies and determine the relative risk of spontaneous delivery before 33 weeks in women with a fibronectin positive result. DESIGN: Prospective clinical study. SETTING: Inner city antenatal clinic. POPULATION: Singleton pregnancies attending for routine antenatal care. METHODS: Cervicovaginal fetal fibronectin and cervical length were measured at 23 weeks of gestation. The distribution of fibronectin positivity within subgroups according to maternal characteristics was calculated and the relative risk of spontaneous delivery before 33 weeks was estimated. MAIN OUTCOME MEASURES: Prevalence of a fibronectin positive result and its relation to cervical length measurement and spontaneous preterm delivery before 33 weeks. RESULTS: Of 5,146 women participating in the study, 182 (3.5%) had a fibronectin positive result and 76 (1.5%) had a cervical length of < 15 mm. Fibronectin positive women were more likely to be Afro-Caribbean in origin, to have had a previous second trimester miscarriage and to have a short cervix. In the 5,068 women who were managed expectantly, the significantly independent relative risk of spontaneous delivery at < 33 weeks was 46.2 (95% CI 18.8-113.6), for cervical length of < or = 15 mm, 8.1 (95% CI 3.8-17.5) for a fibronectin positive result, and 4.4 (95% CI 2.2-9.1) for cigarette smoking. CONCLUSION: Fibronectin positivity at 23 weeks of gestation provides useful prediction of pregnancies at risk of spontaneous preterm delivery before 33 weeks, with a relative risk that is twice as high as cigarette smoking, but is a sixth of that of cervical length. PMID- 11028583 TI - Estimates of maternal mortality by the sisterhood method in rural nothern Tanzania: a household sample and an antenatal clinic sample. AB - OBJECTIVE: To estimate maternal mortality in two samples of a population in northern Tanzania. SETTING: Rural communities and antenatal clinics, Mbulu and Hanang districts, Arusha region, Tanzania. POPULATION: From a household survey 2,043 men and women aged 15-60, and from an antenatal clinic survey 4,172 women aged 15-59. METHOD: The indirect sisterhood method. MAIN OUTCOME MEASURES: The risk of maternal deaths per 100,000 live births (maternal mortality ratio), and the lifetime risk of a maternal death. RESULTS: The risk of a maternal death per 100,000 live births was 362 (95% CI 269-456) and 444 (95% CI 371-517) for the household and antenatal clinic surveys, respectively. The lifetime risk of maternal death was 1 in 38 and 1 in 31, respectively, for the two surveys. A significantly lower risk of maternal death was observed for the respondents attending antenatal clinics closer to the hospital than for those attending clinics further away: 325 (95% CI 237-413) compared with 561 (95% CI 446-677) per 100,000 live births. Lifetime risk of maternal death was 1 in 42 and 1 in 25, respectively. CONCLUSIONS: The risk of maternal death per 100,000 live births in this area were comparatively high, but in our survey substantially lower than in previous surveys in Tanzania. Increasing distance from the antenatal clinics to the hospital was associated with higher maternal mortality. There was no significant difference between results based on household and antenatal clinic data, suggesting that accessible health facility data using the sisterhood method may provide a basis for local assessment of maternal mortality in developing countries. PMID- 11028582 TI - Influenza virus infection in the second and third trimesters of pregnancy: a clinical and seroepidemiological study. AB - OBJECTIVE: To determine whether maternal influenza virus infection in the second and third trimesters of pregnancy results in transplacental transmission of infection, maternal auto-antibody production or an increase in complications of pregnancy. DESIGN: Case-control cohort study. POPULATION: Study and control cohorts were derived from 3,975 women who were consecutively delivered at two Nottingham teaching hospitals between May 1993 and July 1994. A complete set of three sera was available for 1,659 women. METHODS: Paired maternal ante- and postnatal sera were screened for a rise in anti-influenza virus antibody titre by single radial haemolysis and haemagglutination inhibition. Routine obstetric data collected during and after pregnancy were retrieved from the Nottingham obstetric database. Cord samples were tested for the presence of IgM anti-influenza antibodies, and postnatal infant sera were tested for the persistence of influenza-virus specific IgG. Paired antenatal and postnatal sera were tested against a standard range of auto-antigens by immunofluorescence. MAIN OUTCOME MEASURES: Classification of women as having definite serological evidence of an influenza virus infection in pregnancy (cases) or as controls. RESULTS: Intercurrent influenza virus infections were identified in 182/1,659 (11.0%) pregnancies. None of 138 cord sera from maternal influenza cases was positive for influenza A virus specific IgM. IgG anti-influenza antibodies did not persist in any of 12 infant sera taken at age 6-12 months. Six of 172 postnatal maternal sera from cases of influenza were positive for auto-antibodies. In all cases the corresponding antenatal serum was also positive for the same auto-antibody. There were no significant differences in pregnancy outcome measures between cases and controls. Overall, there were significantly more complications of pregnancy in the cases versus the controls, but no single type of complication achieved statistical significance. CONCLUSIONS: Influenza infection in the second and third trimesters of pregnancy is a relatively common event. We found no evidence for transplacental transmission of influenza virus or auto-antibody production in pregnancies complicated by influenza infections. There was an increase in the complications of pregnancy in our influenza cohort. PMID- 11028585 TI - Women's knowledge of and attitudes to first and second trimester screening for Down's syndrome. AB - One hundred women were interviewed at their first hospital antenatal visit to assess their knowledge of, and attitudes to, first versus second trimester screening for Down's syndrome. Overall, the women had limited knowledge of Down's syndrome, and the prenatal screening and diagnostic tests that are available. However, when informed, the majority of women expressed a clear preference for first trimester screening tests for Down's syndrome, regardless of the rate of miscarriage of Down's syndrome pregnancies between 10 and 15 weeks of gestation. These findings have implications for the planning of prenatal Down's syndrome screening programmes. PMID- 11028584 TI - Does histological incomplete excision of cervical intraepithelial neoplasia following large loop excision of transformation zone increase recurrence rates? A six year cytological follow up. AB - OBJECTIVE: To determine the risk of recurrent cervical intraepithelial neoplasia (CIN) in women with complete or incomplete excision of cervical intraepithelial neoplasia treated by large loop excision of transformation zone (LLETZ). DESIGN: A retrospective study SETTING: One consultant-led colposcopy clinic at Leicester Royal Infirmary POPULATION: Three hundred and ninety-four women referred consecutively to the colposcopy clinic between 1991 and 1992. MAIN OUTCOME MEASURES: The histological recurrence rate of CIN, length of cytological follow up following treatment related to degree of completeness of excision at initial treatment. RESULTS: Three hundred and twenty-two women had complete cytological or histological follow up. The mean length of follow up was 73 months with a mean number of six smears. Women with incomplete excision of CIN had a significantly higher risk of recurrent CIN (relative risk 8.23) occurring in a significantly shorter time compared with women with complete excision. CONCLUSIONS: This study demonstrates that large loop excision of transformation zone is successful in over 95% of cases. Cytological surveillance is satisfactory for follow up of women who have complete excision of CIN. Women with incomplete excision of CIN at initial LLETZ remain at significant risk of developing further CIN and long term colposcopic and cytological follow up is necessary. PMID- 11028586 TI - Emotional, physical and sexual violence against women before or during pregnancy. AB - The objective of this study was to determine the prevalence of emotional, sexual or physical violence reported by women delivered at the University Hospital Geneva, a tertiary care university teaching hospital, where about 3,000 women are delivered each year. This study showed that prevalence of violence against women is high, and is severely underestimated by health case providers. PMID- 11028587 TI - A comparison of the contractile properties of human myometrium obtained from the upper and lower uterine segments. AB - This study compared the contractile characteristics of myometrium taken from upper and lower uterine segments. Biopsies were obtained from women undergoing classical caesarean section. Myometrial strips were dissected and mounted in an organ bath, and the contractions were recorded. The cross sectional area of the contractile elements within the strips was measured enabling strips of differing dimensions to be compared. There were no significant differences in the contractile rate and force production produced by myometrium from the upper and lower segments. This study demonstrated that for contractile studies, the use of lower segment is appropriate. The results fail to demonstrate any functional regionality of the human uterus in terms of contractility. PMID- 11028588 TI - Controlled analysis of factors associated with insufficient sample on outpatient endometrial biopsy. AB - We examined the relative significance of hysteroscopic and ultrasonographic evidence of endometrial atrophy in relation to insufficient sample on outpatient endometrial biopsy in women with abnormal uterine bleeding. Multivariate logistic regression modelling was used to evaluate the independent effects of age, menopausal status, hysteroscopic findings and sonographic endometrial thickness on outpatient endometrial sampling (sufficient or insufficient) used as the binary dependent variable. Insufficient sample on endometrial biopsy was associated with hysteroscopic finding of endometrial atrophy (OR 4.79, 95% CI 1.05-21.91, P = 0.04) and sonographic endometrial thickness below 5 mm (OR 0- 19. 95% CI 0.07-0.53, P = 0.001). There was no association with patient's age and menopausal status. In conclusion, when reassuring women with insufficient sample on outpatient endometrial biopsy, one can be confident about absence of pathology provided the hysteroscopic and sonographic endometrial assessment is consistent with endometrial atrophy. PMID- 11028589 TI - Heparin-induced skin necrosis. PMID- 11028590 TI - Limitations of ultrasound in the diagnosis of fetomaternal haemorrhage. PMID- 11028591 TI - Bilateral nipple ulcers in a breastfeeding woman: a manifestation of Behcet's disease? PMID- 11028592 TI - Pelvic tuberculosis: an uncommon gynaecological problem presenting as ovarian mass. PMID- 11028593 TI - A double-blind randomised trial of leuprorelin acetate prior to hysterectomy for dysfunctional uterine bleeding. PMID- 11028594 TI - Parvovirus B19 infection: association with third-trimester intrauterine fetal death. PMID- 11028595 TI - Randomised comparison of Burch colposuspension versus anterior colporrhaphy in women with stress urinary incontinence and anterior vaginal wall prolapse. PMID- 11028596 TI - Management of menorrhagia--SMART study (Satisfaction with Mirena and Ablation: a Randomised Trial). PMID- 11028597 TI - Wrong turn--dead end. PMID- 11028599 TI - High-throughput catalyst screening PMID- 11028598 TI - Synergy in analytical chemistry PMID- 11028600 TI - Search-and-destroy missions in DNA computing. PMID- 11028602 TI - The best of both worlds with LC/FTMS PMID- 11028601 TI - Screening for STDs. PMID- 11028603 TI - Forced flow techniques in planar chromatography PMID- 11028605 TI - Spectrometers get small PMID- 11028604 TI - Polymeric microelectromechanical systems PMID- 11028606 TI - Defining course goals PMID- 11028607 TI - Two-dimensional fluorescence correlation in capillary electrophoresis for peak resolution and species identification. AB - A new spectroscopic dimension-fluorescence intensity correlation--is introduced to enhance peak resolution and species identification in capillary electrophoresis. In two-dimensional correlation CE, a conventional electropherogram is spread into two dimensions through cross-correlation analysis of fluorescence time response. A laser that is sinusoidally modulated in intensity is used as the excitation source. Three channels of information are collected during a CE run: the steady-state intensity, the ac amplitude, and the phase-resolved fluorescence intensity. The correlation between two chosen channels is then evaluated. A two-dimensional correlation electropherogram consists of a plot of the correlation intensity versus two axes of migration time. Through correlation analysis, species discrimination and peak resolution are significantly enhanced without having to physically separate the solutes. Two dimensional correlation CE showed complete resolution between two overlapping sample peaks with a resolution of 0.28 in the conventional one-dimensional electropherogram. In separations of polycyclic aromatic hydrocarbons by micellar electrokinetic chromatography (MEKC), two-dimensional correlation analysis resolved all overlapping elution peaks unseparable by one-dimensional MEKC, demonstrating the utility of 2D correlation in separation method development. The capability of 2D correlation CE in species identification is demonstrated with a sequence of 39 consecutively injected peaks containing four fluorescent dyes. Species identification in sequencing is achieved without complex data treatment in two-dimensional correlation CE. PMID- 11028608 TI - Sequencing of sulfated oligosaccharides from mucins by liquid chromatography and electrospray ionization tandem mass spectrometry. AB - As part of a strategy for profiling diverse mixtures of sulfated mucin-derived oligosaccharides, liquid chromatography coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) in the negative ion mode has been explored. Two mixtures of sulfated oligosaccharide alditols from porcine stomach and large intestine were analyzed by straight phase chromatography using an amino-bonded column connected to a Q-TOF instrument. Nine sulfated mucin-derived oligosaccharide alditols from porcine stomach underwent extensive fragmentation allowing determination of their sequence. The fragmentation generated primary, secondary, and tertiary fragment ions informative for the elucidation of the saccharide sequence and localization of the sulfate group. From a single chromatographic analysis, the sequences of 28 different sulfated mucin oligosaccharide alditols purified from porcine large intestine were elucidated, revealing information concerning prominent core sequences and terminal blood group-type epitopes. Analysis of these two sulfated oligosaccharide mixtures demonstrated the usefulness of HPLC-ESI-MS/MS: the on-line separation of multiple isomeric suffated oligosaccharides as present in biological samples, informative fragmentation allowing the identification of the sequence of nonderivatized oligosaccharides, and a sensitivity sufficient for the analysis of quantities as obtained from natural sources. PMID- 11028609 TI - GPC separation of polymer samples for MALDI analysis AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is an important technique to characterize the average molecular weights, oligomer repeat units, and end groups of polymer materials. Although MALDI methods have been developed for a broad variety of different synthetic polymers, MALDI continues to struggle with polymer samples having broad polydispersity (PD). We have combined MALDI and gel permeation chromatography (GPC) analyses for broad PD polymer samples with the use of a liquid chromatography (LC) interface. The LC interface uses heated sheath gas and a capillary nozzle to remove most of the mobile phase and deposit the GPC eluants on the precoated matrix on a moving MALDI plate. Our experiments demonstrate that the combination of GPC-LC interface MALDI can aid in the characterization of broad PD samples, the verification of the presence of low-intensity, high-mass oligomers, and the detection of minor series in polymer samples. PMID- 11028610 TI - Detection of nine chlorinated and brominated haloacetic acids at part-per trillion levels using ESI-FAIMS-MS AB - A combination of electrospray ionization, high-field asymmetric waveform ion mobility spectrometry, and mass spectrometry (ESI-FAIMS-MS) was used for the analysis of a solution containing a mixture of the nine chlorinated and brominated haloacetic acids. For a carrier gas of nitrogen in the FAIMS analyzer, haloacetate anions of the mono- and dihalogenated acids and the decarboxylated anions of three of the trihalogenated acids were detected. No signal was observed for bromodichloroacetic acid (BDCAA) at a dispersion voltage of -3400 V. The addition of a small amount of carbon dioxide to the nitrogen carrier gas resulted in the detection of the pseudomolecular trihaloacetate anions, including BDCA-, and significant increases in sensitivities for the trihalogenated species. The addition of carbon dioxide to the nitrogen carrier gas had little effect on the mono- and dihalogenated anions. Quantitative analysis of the nine haloacetic acids, using flow injection, gave detection limits between 5 and 36 parts-per trillion in 9/1 methanol/water (v/v) containing 0.2 mM ammonium acetate. PMID- 11028611 TI - Determination of anionic and nonionic surfactants, their degradation products, and endocrine-disrupting compounds in sewage sludge by liquid chromatography/mass spectrometry. AB - A comprehensive analytical method based on reversed-phase liquid chromatography and mass spectrometry using both atmospheric pressure chemical ionization and electrospray ionization has been developed for the simultaneous determination of anionic and nonionic surfactants, their polar degradation products, and endocrine disrupting compounds (EDCs) in sewage sludge. Extraction of target compounds, with recovery rates from 86% to nearly 100% for polyethoxylates and from 84 to 94% for polar degradation products, was achieved applying ultrasonic solvent extraction with a mixture of methanol/ dichloromethane (7:3, v/v). Cleanup of sample extracts was performed on octadecyl solid-phase extraction cartridges. Determination of less polar compounds: alcohol ethoxylates (AEOs), nonylphenol ethoxylates (NPEOs), coconut diethanol amides, poly(ethylene glycol)s, and phthalate esters was accomplished by reversed-phase LC-APCI-MS in positive ionization mode, while more polar compounds: nonylphenolcarboxylates, nonylphenol (NP), octylphenol, and bisphenol Awere analyzed by ion-pair LC-ESI-MS under negative ionization conditions. This protocol was successfully applied to the trace determination of anionic and nonionic surfactants, polar degradation products, and EDCs in sewage sludge collected from different sewage treatment plants. The analysis revealed the presence of NP at high concentration levels ranging from 25 to 600 mg/kg. Polyethoxylates (AEOs and NPEOs) were also found in all samples at parts-per-million levels (10-190 mg/kg AEOs and 2-135 mg/kg NPEOs, respectively). PMID- 11028612 TI - The limitations of MALDI-TOF mass spectrometry in the analysis of wide polydisperse polymers AB - Average molecular weight determination of polymers with polydispersities greater than 1.2 is an ongoing challenge in the field of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Mass discrimination effects observed in the analysis of these polymers have been attributed to sample preparation, desorption/ionization, and instrumental factors. In an effort to separate these factors, we studied poly(methyl methacrylate) (PMMA) standards using two different ion detection systems installed on the same time-of-flight mass analyzer. Equimass blends of narrow PMMA standards were used to simulate a polymer with a wide polydispersity. MALDI-MS analysis was also performed on a PMMA standard with a polydispersity of 1.7. All samples were analyzed by size exclusion chromatography for comparison. Although sample preparation and ionization/desorption factors were found to influence the spectral appearance of the MMA distributions, we demonstrate that, under similar sample preparation and instrument conditions, different ion detection systems produce different results for synthetic polymer blends. The differences in the detector responses for the blends and wide polydisperse standard arise from several factors related to the ion detection system: (1) detection mechanisms, (2) saturation effects, and (3) signal-to-noise limitations. PMID- 11028613 TI - Analysis of a tryptic digest of pig hemoglobin using ESI-FAIMS-MS. AB - The continuous gas-phase ion separation and atmospheric pressure focusing properties of high-field asymmetric waveform ion mobility spectrometry (FAIMS) offer significant advantages for the mass spectrometric analysis of tryptic digests of proteins. In this study, tryptic peptides of pig hemoglobin were examined by ESI-FAIMS-MS using a newly designed FAIMS device. The new, hemispherical geometry of the inner electrode served to deliver the ions, via the gas flows, to the center axis of the FAIMS analyzer, improving the sensitivity relative to previous prototypes. Mass spectra collected using this new FAIMS showed significantly less chemical background noise than conventional ESI-MS, while maintaining approximately the same absolute sensitivity as that observed with ESI-MS. As a consequence of the ion separation in FAIMS, the identification of the tryptic fragments was simplified and some peptides, such as the triply protonated WAGVANALAHK3+, that were obscured by the intense background of ESI-MS, were readily detected using ESI-FAIMS-MS. In addition, the FAIMS device was shown to separate isobaric ions at m/z 532.4. Correlations between CV and mass-to charge ratio, as well as CV and ionic collision cross section, were evaluated for 38 peptide ions identified in the tryptic digest. The correlation between the CV of the peptide and the mass-to-charge ratio is very poor, indicating good orthogonality between the separation by FAIMS and the separation by mass spectrometry. PMID- 11028614 TI - Interchain ion formation in secondary ion mass spectrometry resulting from the double-helical structure of isotactic poly(methyl methacrylate) in adsorbed monolayers AB - Results from time-of-flight secondary ion mass spectra (TOF-SIMS) of Langmuir Blodgett monolayers of various isomers (isotactic and syndiotactic) of poly(methyl methacrylate) (PMMA) are reported. A detailed analysis of the repeating pattern of fragment ion clusters yields very different patterns for isotactic PMMA LB layers than for the syndiotactic and atactic forms. This is attributed to the resulting double-helical tertiary structure of isotactic PMMA, a structure that does not form for the syndiotactic and atactic PMMA polymer monolayers. The double-helical structure of isotactic PMMA monolayers is verified using reflection absorption Fourier transform infrared spectroscopy. The repeating patterns of cluster ions in syndiotactic and atactic PMMA monolayers can be explained using statistical chain-breaking models for fragmentation of single isolated polymer chains. The repeating ion patterns from the TOF-SIMS of the isotactic PMMA monolayers are analyzed by considering bond breaking and ion formation between adjacent polymer chains, resulting in a newly proposed ion formation model due to the tertiary structure of the double-helical form. A rearrangement mechanism consistent with all ions that are formed is proposed. PMID- 11028615 TI - MALDI-TOF mass spectrometry of insoluble giant polycyclic aromatic hydrocarbons by a new method of sample preparation AB - The insolubility of giant polycyclic aromatic hydrocarbons (PAHs) prevents their characterization by conventional analytical methods, which require a solubilization of the analyte. Laser desorption mass spectrometry may be used to analyze insoluble samples but is limited to relatively low molecular weights (approximately 2000), in the case of PAHs. To overcome this limitation, we applied MALDI-TOF mass spectrometry. Since MALDI sample preparation also requires solubility of analyte and matrix molecules, the sample preparation needed modification. The giant PAHs (>2000 Da) were investigated after using a new sample preparation, consisting of mechanically mixing analyte and matrix without any solubilization procedures. This solvent-free process allows insoluble compounds to be characterized. Furthermore, new organic molecules can be used as a matrix. Indeed, 7,7,8,8-tetracyanoquinodimethane, a new matrix with promising properties, has proven to be particularly suitable for the measurement of PAHs. Thanks to the successful characterization with MALDI-TOF mass spectrometry, the chemical design of giant PAHs, which was hindered until now for a lack of analytical methods, can now continue to develop. PMID- 11028616 TI - Injection of fluorescently labeled analytes into microfabricated chips using optically gated electrophoresis AB - Optically gated electrophoresis has been used as an alternative method of sample introduction in microfabricated chips. Utilization of this injection technique permits rapid serial sampling and consumes less chip space than traditional chip based injection methods. 4-Chloro-7-nitrobenzofurazan (NBD)-labeled amino acids have been injected in microfabricated chips using optical gating and have yielded results comparable to the T-type injection methods currently used. Picoliter-size injection volumes have been reproducibly introduced with less than 3% deviation in retention time and peak area. Six consecutive separations have been accomplished in under 30 s using a separation length of 1.1 cm. Plate heights from this analysis range from 4.5 to 0.8 microm. The combination of rapid successive injections and reproducible separations demonstrates the utility of this method in analyzing rapid dynamic events such as on-line sampling or monitoring chemical processes. PMID- 11028617 TI - Capillary isoelectric focusing of yeast cells. AB - In the present work, capillary isoelectric focusing (CIEF) methods were developed for the separation and identification of yeast cells. Yeast cells (approximately 4-microm diameter) cultured to various phases of growth were shown to be reproducibly resolved by CIEF using 100-microm-i.d. fused-silica capillaries coated with hydroxypropyl methylcellulose. Separation efficiencies corresponding to peak capacities of >4000 were obtained. The suitable cell concentration range for obtaining repeatable elution in CIEF separations was found to be quite low (<3 cells/microL). CIEF experiments showed that yeast cell populations at early log, mid log, and stationary growth phases differ in isoelectric point, with values ranging from 5.2 to 6.4. The broader application of CIEF are projected for microorganism identification and separation based upon growth conditions. PMID- 11028618 TI - Electrophoretic separation of proteins on a microchip with noncovalent, postcolumn labeling. AB - Proteins were separated by microchip capillary electrophoresis and labeled on chip by postcolumn addition of a fluorogenic dye, NanoOrange, for detection by laser-induced fluorescence. NanoOrange binds noncovalently with hydrophobic protein regions to form highly fluorescent complexes. Kinetic measurements of complex formation on the microchips suggest that the reaction rate is near the diffusion limit under the conditions used for protein separation. Little or no band broadening is caused by the postcolumn labeling step. Lower limits of detection for model proteins, alpha-lactalbumin, beta-lactoglobulin A, and beta lactoglobulin B, were <0.5 pg (approximately 30 amol) of injected sample. The relative fluorescence and reaction rates are compared with those of a number of other fluorogenic dyes used for protein labeling. PMID- 11028619 TI - Chiral monolithic columns for enantioselective capillary electrochromatography prepared by copolymerization of a monomer with quinidine functionality. 1. Optimization of polymerization conditions, porous properties, and chemistry of the stationary phase. AB - Monolithic columns for chiral capillary electrochromatography have been prepared within the confines of untreated fused-silica capillaries in a single step by a simple copolymerization of mixtures of O-[2-(methacryloyloxy)ethylcarbamoyl] 10,11-dihydroquinidine , ethylene dimethacrylate, and glycidyl methacrylate or 2 hydroxyethyl methacrylate in the presence of mixture of cyclohexanol and 1 dodecanol as a porogenic solvent. The porous properties of the monolithic columns can easily be controlled through changes in the composition of the binary porogenic solvent. Although both thermal- and UV light-initiated polymerizations afford useful capillary columns, monoliths prepared using the former approach exhibit better chromatographic properties. The ability to control pore size independently of the polymerization mixture composition enables the preparation of monoliths with varying percentages of the chiral monomer and cross-linker, as well as the optimization of their separation properties. Very good separations of model racemate (R,S)-N-3,5-dinitrobenzoylleucine were achieved using an optimized monolithic CEC column, with high efficiencies of up to 74000 plates/m for the retained peaks. PMID- 11028620 TI - Chiral monolithic columns for enantioselective capillary electrochromatography prepared by copolymerization of a monomer with quinidine functionality. 2. Effect of chromatographic conditions on the chiral separations. AB - The effect of chromatographic conditions on the performance of chiral monolithic poly(O-[2-(methacryloyloxy)-ethylcarbamoyl]-10,11-dihydroqui nidine-co-ethylene dimethacrylate-co-2-hydroxyethyl methacrylate) columns in the capillary electrochromatography of enantiomers has been studied. The flow velocity was found to be proportional to the pore size of the monolith and both the pH and the composition of the mobile phase. The length of both open and monolithic segments of the capillary column was found to exert a substantial effect on the run times. The use of monoliths as short as 8.5 cm and the "short-end" injection technique enabled the separations to be achieved in approximately 5 min despite the high retentitivity of the quinidine selector. Very high column efficiencies of close to 250000 plates/m and good selectivities were achieved for the separations of numerous enantiomers using the chiral monolithic capillaries with the optimized chromatographic conditions. PMID- 11028621 TI - Mixed-mode anion-cation exchange/hydrophilic interaction liquid chromatography electrospray mass spectrometry as an alternative to reversed phase for small molecule drug discovery AB - Within pharmaceutical drug discovery, significant needs currently exist for the analysis and purification of structurally diverse samples prior to or immediately following high-throughput screening. These processes are required to facilitate rapid and accurate biological profiling, structural determination, and resupply of new drug candidates. Reversed-phase high-performance liquid chromatography (RP HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) for both analytical and preparative applications has become the small molecule separation/detection tool of choice for meeting many of these needs. However, the separation selectivity provided by RP-HPLC has been limited to the hydrophobicity based resolution of relatively nonpolar sample components, and for high throughput drug discovery applications, no sufficient alternative procedures have been identified. In this investigation, a mixed-mode anion-cation exchange/hydrophilic interaction chromatography (ACE-HILIC) method has been developed to provide both direct compatibility with ESI-MS and evaporative light scattering detection (ELSD) and separation selectivity highly orthogonal to RP HPLC. The technique employed silica-based small-pore weak ion exchange resins eluted with a combined aqueous and pH gradient. A diverse set of dipeptide probes was employed for the elucidation of the relative contributions of three retention mechanisms. ACE-HILIC-ESI-MS-ELSD should prove useful for the analysis and purification of compounds from both biological (e.g., natural products) and synthetic (e.g., combinatorial chemistry) sources of molecular diversity. PMID- 11028622 TI - Affinity capillary electrophoresis for the assessment of complex formation between viruses and monoclonal antibodies. AB - The formation of complexes of human rhinovirus (serotype HRV2 and HRV14) with nonaggregating neutralizing monoclonal antibodies was investigated by affinity capillary electrophoresis. The method is based on preincubation of virus with antibody, followed by CE analysis. At low antibody-to-virus ratios, peaks corresponding to the complexes were broad, pointing to the presence of a heterogeneous population of virions with various numbers of antibodies bound; at a high molar ratio between virus and antibody, the peak became narrow again, indicating saturation of the 60 equivalent viral epitopes with the antibodies being attached bivalently. As SDS was used as an additive in the background electrolyte to allow for separation, its influence on complex formation was investigated. Once formed, HRV2-antibody complexes were found to be stable in the presence of the detergent but complex formation in buffer containing SDS was severely impaired. HRV14-antibody complexes were rapidly dissociated by SDS. The method proved to be useful for a rapid assessment of complex formation and might allow for an estimation of the binding stoichiometry. PMID- 11028623 TI - High-throughput single-molecule spectroscopy in free solution. AB - A high-speed high-throughput single-molecule imaging technique for identifying molecules in free solution based on differences in their fluorescence emission spectra is presented. Unlike previous reports, the entire spectrum, rather than selected wavelengths through optical filters, is recorded. Furthermore, the millisecond data acquisition time means that the molecules do not need to be immobilized or spatially confined. In one example, individual lambdaDNA molecules labeled with YOYO-I, POPO-III, or a combination of the two dyes can be distinguished from one another. In another example, biotinylated 2.1-kb DNA labeled with YOYO-I was reacted with avidin-conjugated R-phycoerythrin. The two different reactant molecules and the product molecule can be simultaneously imaged and identified by their spectroscopic characteristics. This technique can therefore be used for screening single molecules for disease markers and for monitoring individual molecular interactions at a rate of thousands of molecules per second. PMID- 11028624 TI - Orientations of liquid crystals on mechanically rubbed films of bovine serum albumin: a possible substrate for biomolecular assays based on liquid crystals. AB - We report the uniform planar anchoring of thermotropic liquid crystals on films of bovine serum albumin (BSA) covalently immobilized on the surface of glass microscope slides and mechanically rubbed using a cloth. The azimuthal orientations of the liquid crystals were measured to be parallel to the direction of rubbing. Following immersion and removal of these rubbed films of BSA from aqueous solutions containing either BSA, fibrinogen, lysozyme, anti-FITC immunoglobulin G (IgG), or antistreptavidin IgG, we measured liquid crystals placed onto these surfaces to largely retain their uniform alignment. In contrast, following immersion of a rubbed film of BSA into an aqueous solution of anti-BSA IgG, we observed liquid crystals on these surfaces to assume nonuniform orientations. We conclude that specific binding of anti-BSA IgG to the film of rubbed BSA erased anisotropy induced within the film of BSA by rubbing. This result suggests that the spatial scale of anisotropy within the rubbed film of BSA is comparable to or smaller than the size of the IgG molecule. Because the anisotropy within a rubbed film of a protein can be erased by specific binding of a second protein, we believe these types of substrates (rubbed films of proteins) have the potential to be useful in a variety of label-free biomolecular assays where specific binding of a target species to its ligand can be imaged through observation of the optical appearance of liquid crystal placed onto the surface. PMID- 11028625 TI - Trace analysis of ethanol, MTBE, and related oxygenate compounds in water using solid-phase microextraction and gas chromatography/mass spectrometry. AB - Solid-phase microextraction (SPME) and gas chromatography/mass spectrometry have been combined for trace-level determination of very polar compounds in water, including the widely used gasoline oxygenates ethanol and methyl tert-butyl ether (MTBE). A relatively simple extraction method using a divinylbenzene/Carboxen/poly(dimethylsiloxane) SPME fiber was optimized for the routine analysis of ethanol and MTBE in groundwater and reagent water. A sodium chloride concentration of 25% (w/w) combined with an extraction time of 25 min provided the greatest sensitivity while maintaining analytical efficiency. Replicate analyses in fortified reagent and groundwater spiked with microgram per liter concentrations of ethanol and MTBE indicate quantitative and reproducible recovery of these and related oxygenate compounds. Method detection limits were 15 microg L(-1) for ethanol, 1.8 microg L(-1) for tert-butyl alcohol, 0.038 microg L(-1) for tert-amyl methyl ether, 0.025 microg L(-1) for ethyl-tert-butyl ether, and 0.008 microg L(-1) for MTBE. PMID- 11028626 TI - Analysis of pesticides by LC-electrospray-MS with postcolumn removal of nonvolatile buffers. AB - Liquid chromatographic (LC) separations for pesticides and many other compounds make use of nonvolatile buffers in the mobile phase. The coupling of LC with mass spectrometry (MS) does not allow the use of nonvolatile buffers. Substitution with volatile buffers is possible, but changes in chromatographic retention and resolution can result even if pH is held constant. The postcolumn removal of nonvolatile buffers using a commercially available ion suppressor is evaluated for the analysis of carbamate pesticides. The suppressor efficiently removes phosphate anions from an LC mobile phase. Most compounds show an increased signal by factors of 2-7 after postcolumn phosphate removal. The suppressor has little effect on the chromatographic parameters of some compounds, while serious negative effects are noted for others. Some compounds will give poor results due to adsorption or retention by the suppressor. The results indicate that such a device may be useful for the LC-MS analysis of some pesticides using nonvolatile buffers. PMID- 11028627 TI - Constant holdup times in gas chromatography by programming of column temperature and inlet pressure AB - The requirements for attaining constant holdup times in capillary gas chromatography (GC), while linearly programming both column temperature and inlet pressure, are illustrated. A scheme, which is based on a series of responses from isothermal and isobaric conditions, is proposed. The responses from two commonly used GC instruments, one with a mass-selective detector operating at near-vacuum pressure and another with a flame-ionization detector at ambient pressure, are analyzed. A significant deviation from Poiseuille flow is noted due to the use of helium as a carrier gas. Nonetheless, the experimental holdup times are approximately constant over a range of temperatures and pressures. Theoretical analysis reveals the spatial and temporal dependence of flow rates inside the column during the programmed runs. PMID- 11028628 TI - The design of cyclic AMP--recognizing oligopeptides and evaluation of its capability for cyclic AMP recognition using an electrochemical system. AB - A novel 17-mer peptide ligand for cyclic AMP was designed using the amino acid sequences of essential subsites in various cyclic AMP-dependent protein kinase (protein kinase A) families. The Au disk electrode, which was modified with the designed 17-mer oligopeptide, responded to cyclic AMP but virtually did not respond to any other cyclic nucleotides using the ion channel sensor mechanism. On the other hand, a scrambled peptide, which had the same amino acid composition as and had an amino acid sequence different from the 17-mer oligopeptide, did not respond to any nucleotides. This indicates that the designed 17-mer peptide actually acted as a selective ligand for cyclic AMP. This ligand-designing strategy using peptide sequences in target-binding proteins may possibly be extended to the design of peptide ligands for other second messengers. PMID- 11028629 TI - Improved computation of the standard error in the regression coefficient estimates of a multivariate calibration model AB - A multivariate calibration model consists of regression coefficient estimates whose significance depends on the associated standard errors. A recently introduced leave-one-out (LOO) method for computing these standard errors is modified to achieve consistency with the jack-knife method. The proposed modification amounts to multiplying the LOO standard errors with the factor (n - 1)/n1/2, where n denotes the number of calibration samples. The potential improvement for realistic values of n is illustrated using a practical example. PMID- 11028630 TI - Electrochemical detectors prepared by electroless deposition for microfabricated electrophoresis chips AB - Microfabricated capillary electrophoresis (CE) chips with integrated electrochemical detection have been developed on glass substrates. An electroless deposition procedure was used to deposit a gold film directly onto the capillary outlet to provide high-sensitivity electrochemical detection for catechol and several nitroaromatic explosives. Scanning electron microscopy revealed that the electroless gold film contains nanoscopic gold aggregates (100-150 nm) with an average thickness of 79 nm. The electroless deposition procedure can be easily and routinely performed in any wet-chemistry laboratory, and electroless gold can be deposited onto complex and internal surfaces. Intimate coupling of electrochemical detection and CE chips obviates the need for a coupling mechanism or tedious alignment procedures. With nitroaromatic compounds as a working model, microchip capillary electrophoresis equipped with electroless gold has proven to provide high sensitivity and fast response times for sensor applications. The CE microchip system was capable of separation and determination of explosive compounds including TNT in less than 130 s with detection limits ranging from 24 to 36 microg/L, i.e., 4-fold enhancements in detection efficiency in comparison to thick-film technology. PMID- 11028631 TI - Thiazole-containing benzo-crown ethers: a new class of ammonium-selective ionophores AB - A new class of crown ethers containing heterocyclic units has been designed to develop synthetic NH4+-selective ionophores whose resulting potentiometric properties in solvent polymeric membranes are superior or at least comparable to those of nonactin. It was found that the derivatives of thiazole containing dibenzo-18-crown-6 (1; TDB18C6) incorporated in PVC-based membranes provide enhanced NH4+ selectivities over alkali metal cations, especially over Na+, compared to those of the nonactin-doped membranes: for example, the selectivity coefficients, log K(POT)NH4+J (J = Li+, Na+, K+), for the 2-nitrophenyloctyl ether (NPOE) plasticized PVC membranes doped with the hexyl chain-substituted TDB18C6 (2) were -4.9, -3.9, and -1.3, while those for the same type of membranes with nonactin -4.4, -3.0, and -1.0, respectively. Unlike other synthetic NH4+ selective neutral carriers reported to date, TDB18C6-type compounds result in potentiometric performance highly comparable to that of the nonactin-based ones except their slightly higher detection limits (approximately 3 x 10(-6) vs 7 x 10(-7)). The complex formation between TDB18C6 and NH4+ was identified from 1H NMR spectra. The 1:1 complex formation constant of TDB18C6 with NH4+ in solvent polymeric membranes estimated using the method suggested by Bakker et al. and the NH4+ selectivity coefficient over K+ (K(POT)NH4+,K+) are approximately 200 times smaller than that of nonactin (1.8 x 10(7) vs 3.6 x 10(9)). PMID- 11028632 TI - Extended-range glucose sensor employing engineered glucose dehydrogenases. AB - An enzyme glucose sensor with an expanded dynamic range was constructed using a novel strategy. This strategy was based on a new concept of utilizing protein engineered enzymes with a different Michaelis constant, which allows for the expanded dynamic range. We used the engineered Escherichia coli pyrroloquinoline quinone glucose dehydrogenase (PQQGDH) of which His775 was substituted for Asp which showed an increased Km value (25-fold). We first constructed the composite colorimetric analytical system employing the wild-type PQQGDH and His775Asp and evaluated its dynamic range. The composite colorimetric analytical system was constructed and showed a wide dynamic range of 0.5-30 mM with less than +/-5% error. The composite colorimetric analytical system, an extended-range colorimetric analytical system, enabled the determination of the concentration of glucose over a 30-fold range that could not have been achieved using the single colorimetric analytical system. Furthermore, we have demonstrated the composite amperometric glucose sensor employing the combination of His775Asn and His775Asp. The extended-range glucose sensor acquired not only the expanded dynamic range (3 70 mM) that covered both dynamic ranges of the single enzyme sensors but also the narrower substrate specificity of glucose due to the inherited property of engineered enzymes. PMID- 11028633 TI - Potentiometric evaluation of solvent polymeric carbonate-selective membranes based on molecular tweezer-type neutral carriers AB - Potentiometric properties of the ion-selective electrodes) based on highly plasticized PVC membranes doped with the carbonate-selective cholic acid (CA) derivatives have been measured. The carbonate-selective neutral carriers have been prepared by coupling one to three trifluoroacetobenzoyl (TFAB) groups to a cholic acid derivative which has three hydroxyl linkers lining on the C3, C7, and C12 positions of its rigid steroidal ring structure. The membranes based on cholic acid derivatives with two TFABs [3,7-bis(TFAB)CA, 3,12-bis(TFAB)CA, and 7,12-bis(T-FAB)CA] exhibited remarkably improved carbonate selectivity, indicating that the bis(FAB)CAs behave like molecular tweezers for the carbonate ion. For example, 3,12-bis(TFAB)CA resulted in 10-300-fold-enhanced carbonate selectivity over other anions (e.g., salicylate, ClO4-, SCN-, (HPO4)2-, NO3-, NO2 , Br-, and Cl-) compared to that of the neutral carriers with a single TFAB group. The distances between the carbonate binding centers of bis(TFAB)CAs, i.e., the carbonyl carbons of the two TFAB groups, are in the 7.3-7.9-A range at the AM1 level semiempirical calculation, which is too far for the carbonate ion to form direct covalent bonding. The fast atom bombardment mass spectra of bis(TFAB)CAs show that significant fractions of the compounds are either mono- or dihydrated before complexing the carbonate ion. These findings seem to suggest that bis(TFAB)CAs recognize the incoming carbonate ion by forming both covalent and hydrogen bonding between the hydrated and unhydrated TFAB groups. The analytical utility of the carbonate-selective electrode based on 3,12-bis(TFAB) CA has been demonstrated by measuring the total carbon dioxide in human serum in the presence of lipophilic anion interferents, e.g., salicylate. PMID- 11028634 TI - Interactions of benzyl viologen with surface-bound single- and double-stranded DNA. AB - The interactions of benzyl viologen (BV) with single- and double-stranded calf thymus DNA immobilized onto gold electrodes have been studied by electrochemical methods. Benzyl viologen interacts electrostatically with both double-stranded (ds) and single-stranded (ss) DNA, and the strength of the interactions is dependent on ionic strength (mu). The dicationic form (BV2+) binds to dsDNA 9 times more strongly than the singly reduced form, BV*+, in a pH 7.4 Tris-HCl buffer solution at mu = 8.4 mM. BV2+ binds to ssDNA 5 times more strongly than the BV*+ form. From measurements at mu = 8.4 mM, a binding constant (K2+) of 2.0 (+/-0.2) x 10(4) M(-1) and a binding site size (s) of 1 base pair were obtained, respectively, for dsDNA. For ssDNA, at the same ionic strength, the values obtained for K and s were 3.6 (+/-0.4) x10(4) M(-1) and 2 nucleotides, respectively. The amount of BV bound, whether to dsDNA or ssDNA, decreased with increasing ionic strength. Whereas the binding rate of BV to both dsDNA and ssDNA immobilized onto gold electrodes is relatively low, once immobilized, it dissociates rapidly away from the electrode surface. The electron-transfer rate constant for BV is moderately fast at both dsDNA- and ssDNA-modified gold electrodes. The application of benzyl viologen as an electroactive indicator capable of differentiating between surface-immobilized single- and double stranded DNA in denaturation/regeneration cycles has been explored. PMID- 11028635 TI - Probing trace phenols based on mediator-free alumina sol--gel-derived tyrosinase biosensor. AB - A novel tyrosinase biosensor has been developed for the subnanomolar detection of phenols, based on the immobilization of tyrosinase in a positively charged Al2O3 sol-gel membrane on a glassy carbon electrode. It has been found that Al2O3 sol gel is perfectly beneficial to the immobilization of tyrosinase, because it not only possesses the general advantages of sol-gel but it also is an effective promoter of the biosensor. The large microscopic surface area, porous morphology, and hydrophilic property of the sol-gel matrix result in high enzyme loading, and the enzyme entrapped in this matrix retains its activity to a large extent. The Al2O3 sol-gel-containing surface also displays an intrinsic electrocatalytic o quinone response and, hence, offers a high-sensitivity (127 microA mM(-1)) monitoring of phenols. The detection limit is 0.2 nM at a signal-to-noise ratio of 3, the response time is less than 4 s reaching 95% of the steady-state value, and 70% of the activity is retained after 3 months. PMID- 11028636 TI - Continuous on-line true titrations by feedback-based flow ratiometry. The principle of compensating errors AB - We introduce a new concept for continuous on-line titrations based on feedback controlled flow ratiometry and the principle of compensating errors. The system has been thoroughly tested by applying it to acid-base neutralization titrations with indicator-based end point detection. In a typical case, the total flow (FT, consisting of the sample and the titrant flows) is held constant while the titrant (e.g., a standard base containing an indicator) flow FB varies linearly in response to a controller output voltage. The sample (e.g., an acidic solution to be titrated) flow FA constitutes the makeup and thus also varies (FA = FT - FB). The status of the indicator color in the mixed stream is monitored by an optical detector and used either for governing the controller output or for interpreting the results of the titration. Three methods (PID based control, fixed triangular wave control, and feedback-based triangular wave control implemented on a PC) were examined. In the last and the most successful approach, the titrant flow is initially ramped upward linearly. At the instant a change in the color is sensed by the detector, the titrant flow rate FH is higher than the true equivalence flow rate FE because of the lag time between the first compositional change and its detection. The sensing of the change in color causes the system output to immediately reverse its ramp direction such that the titrant flow now goes down linearly at the same rate. At the instant a change in color, in the opposite direction this time, is again sensed, the titrant flow rate FL is lower than FE by exactly the same amount that FH was higher than FE. This principle of compensating errors (FE = (FH + FL)/2) allows true titrations with excellent reproducibility and speed (0.6% RSD at 3 s/titration and 0.2% RSD at 10 s/titration) and titrant volume consumption as little as 12 microL/titration and solves an old conceptual problem in flow based titrations. PMID- 11028637 TI - Investigation of the molecular extraction process in single subpicoliter droplets using a near-infrared laser Raman trapping system AB - The near-infrared (NIR) laser Raman trapping system was applied to study liquid liquid extraction in a single droplet in a subpicoliter range. The system trapped a single subpicoliter toluene droplet in water using the NIR laser beam and provided time-dependent optical images of the droplet during liquid-liquid extraction. The size of the trapped droplet gradually increased afterp nonylphenol solution was added in water. The Raman spectra of the droplet prove that the increase of the droplet size was caused by the absorption of p nonylphenol from the water. The distribution coefficient of p-nonylphenol in the sub-picoliter droplet was much higher than that in bulk solution. PMID- 11028638 TI - Heat index flow monitoring in capillaries with interferometric backscatter detection AB - Independent control of fluid is important for small volume fluid applications including separation science techniques and analytical microchip devices. To effectively control fluid, a monitoring system is necessary to ensure proper response to the control input. Proof of principle for a new online technique for flow monitoring is demonstrated here. It offers near real-time measurements of small-volume flows in a noninvasive, simple, and robust design. The measurement is accomplished by the introduction of a plug of heat into the fluid and detection of the time of its appearance a short distance downstream. Using heat indexing and laser induced interferometric backscatter as the detector, flow monitoring is demonstrated in 184-microm diameter capillary tubing over a range of 2-25 cm/s (500 nL/s to 7 mL/s.). PMID- 11028639 TI - Topographical and chemical microanalysis of surfaces with a scanning probe microscope and laser-induced breakdown spectroscopy AB - Spatially resolved chemical imaging is achieved by combining a fiber-optic scanning probe microscope with laser-induced breakdown spectroscopy in a single instrument, TOPOLIBS. Elemental composition of surfaces can be mapped and correlated with topographical data. The experiment is conducted in air with minimal sample preparation. In a typical experiment, surface topography is analyzed by scanning a sharp fiber-optic probe across the sample using shear force feedback. The probe is then positioned over a feature of interest and pulsed radiation is delivered to the surface using a nitrogen laser. The pulse vaporizes material from the surface and generates a localized plasma plume. Optical emission from the plume is analyzed with a compact UV/visible spectrometer. Ablation crater size is controlled by the amount of laser power coupled into the probe. Sampling areas with submicrometer dimensions are achieved by using reduced laser power. PMID- 11028640 TI - Voltammetric measurement in supercritical CO2 utilizing a water-in-CO2 microemulsion AB - A novel method for voltammetric measurement in supercritical CO2 is described. In this method a water-in-supercritical CO2 microemulsion is utilized to raise the conductivity of the fluid phase. Well-defined voltammograms for the redox reactions of ferrocene and tetramethyl-p-phenylenediamine were obtained in supercritical CO2 in the presence of the microemulsion. The electrolysis product is stabilized in the water core of the microemulsion. This implies a wide range of electrochemical synthesis involving ionic or radical species in supercritical CO2 may be possible utilizing the microemulsion. The electrochemical results obtained from the water-in-supercritical CO2 microemulsion system are different from that obtained from a supercritical CO2-ethanol mixture system. PMID- 11028641 TI - Determination of alpha- and beta-hydroxycarbonyls and dicarbonyls in snow and rain samples by GC/FID and GC/MS employing benzyl hydroxyl oxime derivatization AB - A flame ionization detector (FID) combined with capillary gas chromatography (GC/FID) and gas chromatography/ mass spectrometry (GC/MS) has been used to identify multifunctional carbonyls in wet precipitation samples. The carbonyl groups were first derivatized to O-benzylhydroxyloxime (BH oxime) by using O benzylhydroxylamine. The BH oxime derivatives were then treated with N,O bis(trimethylsilyl)acetamide for the hydroxyl group to derive their TMS ethers. The BH oxime/TMS derivatives were measured using GC/FID as well as GC/MS on positive EI and CI (isobutane was used as CI gas) modes. Three groups of carbonyl compounds (monoaldehydes, dicarbonyls, hydroxycarbonyls) were identified in the samples by using this method. We have identified, for the first time, a group of alpha- and beta-hydroxycarbonyls, glycolaldehyde, hydroxyacetone, and 4-hydroxy-2 butanone, in wet precipitation samples. Concentrations of hydroxycarbonyls ranged from 0.9 to 53.8 microg/L in the precipitation samples. Their concentration level is similar to that of low molecular weight dicarboxylic acids, which have been reported as major water-soluble organic compounds in rain. PMID- 11028642 TI - Accuracy in the determination of isoelectric points of some proteins and a peptide by capillary isoelectric focusing: utility of synthetic peptides as isoelectric point markers. AB - To evaluate the accuracy ofisoelectric point determination by capillary isoelectric focusing, the pI values of nine proteins and a peptide, the pI values of which had been determined by other methods and ranging pI 3.55-9.60, were determined by capillary isoelectric focusing by cofocusing of recently developed peptide pI markers ranging 3.38-10.17, and the consistency of the pI values was examined. Isoelectric focusing was carried out in neutral polymer-coated capillaries, and the pH gradient was mobilized by pressure toward the cathode, to detect samples with absorption at 280 nm at a fixed detection point. Carrier ampholytes from two different suppliers and in different pH ranges were used. The sharp peaks of the highly pure peptide pI markers greatly facilitated the unambiguous identification of the peaks. When a carrier ampholyte ranging over the acidic side was used, the detection of acidic pI samples was anomalously delayed. This could be partly mitigated by reducing the viscosity of the anode solution in comparison with the pH gradient formed in the capillary. Since the detection times vs the pH relationships were not linear in most cases, the use of a linear calibration line over an entire pH gradient would be erroneous. Instead, the pI values of samples were calculated by assuming a linear relation for pH against detection time between two flanking marker peptides. Close agreement between the pI values, determined by capillary isoelectric focusing, and the reference values of the samples was observed within an average difference range of 0.04-0.08 pH unit with a sample consumption of 10-100 ng within 30-60 min. Some carrier ampholytes were preferentially more effective at either the acidic or the basic side of the pH gradient. For confirmation of the completion of focusing, the use of two different focusing times is recommended. PMID- 11028644 TI - Calibration of analytical instruments. Impact of nonconstant variance in calibration data PMID- 11028643 TI - Capillary isoelectric focusing without carrier ampholytes. PMID- 11028645 TI - Health care costs of smoking. PMID- 11028646 TI - Antidepressants for smoking cessation: a promising new approach? PMID- 11028647 TI - Drug history and remote exposure to drugs. A cause of lung disease? PMID- 11028648 TI - The economic impact of smoking in Germany. AB - Smoking is a high-risk behaviour affecting health and economic welfare of society. Thus it is important to quantify the economic burden smoking places on social institutions in Germany. Approximately 33.4% of the male and 20.4% of the female population are current smokers. This study investigates the health care costs of smoking based on 1996 figures, focusing on the seven most frequent diseases associated with the inhalation of tobacco smoke: chronic obstructive pulmonary disease (COPD, international classification of diseases (ICD) 490-491); lung cancer (ICD 162); stroke (ICD 434-438); coronary artery disease (ICD 410 414); cancer of the mouth and larynx (ICD 140-149, 161) and artherosclerotic occlusive disease (ICD 440). A data search was carried out on MEDLINE, the German Institute for Medical Documentation and Information, and the Internet as well as in databases of health insurance companies and the German Federal institute of statistics. Direct and indirect costs were calculated separately. The results estimate the total smoking related health care costs (attributable fraction due to smoking) for COPD to be 5.471 billion EURO (73%), for lung cancer 2.593 billion EURO (89%), for cancer of the mouth and larynx 0.996 billion EURO (65%), for stroke 1.774 billion EURO (28%), for coronary artery disease 4.963 billion EURO (35%) and for atherosclerotic occlusive disease 0.761 billion EURO (28%). The economic burden of smoking related health care costs for Germany is 16.6 billion EURO. Smoking is therefore responsible for 47% of the overall costs of these diseases (35.2 billion EURO). In the view of the high costs for smoking, of which almost 50% are due to respiratory disease, pneumologists should enhance their effort in primary, secondary and tertiary prevention. PMID- 11028649 TI - Daily respiratory mortality and PM10 pollution in Mexico City: importance of considering place of death. AB - Significant associations have been reported between particles with a 50% cut-off aerodynamic diameter of 10 mm (PM10) and ozone ambient concentrations, and daily number of deaths from respiratory causes. The aim of the present study was to assess such associations among elderly (> or =65 yrs) residents of Mexico City. Ambient air pollution data were provided by the Metropolitan Monitoring Network. During the study period, the average daily PM10 ranged 23.4-175.3 microg x m(-3), and ozone 1 h daily maximums ranged 39.4-216.7 ppb. Information was compiled on the primary and underlying causes of death. The analyses were conducted separately according to place of death (within or out of a hospital unit) using time-series methodology. The total number of deaths from all respiratory causes and mortality for chronic obstructive pulmonary diseases (COPD) were significantly related to PM10 over different lags: an increase of 10 microg x m( 3) was related to a 2.9% (95% (CI): 0.9-4.9%) increase and to a 4.1% (95% CI: 1.3%-6.9%) increase with a 3-day lag when death occurred out of medical units, respectively. For deaths occurring in medical units, a longer lag and smaller risk estimate was observed. An interactive effect between PM10 and ozone was detected. This study confirms that there is an important impact of PM10 on respiratory morbidity among elderly subjects. It also indicates that accounting for primary and underlying causes of death, and considering place of death may reduce misclassification and provide more accurate estimates of the adverse impact of PM10 on mortality. PMID- 11028650 TI - Interaction of vitamin C with the relation between smoking and obstructive airways disease in EPIC Norfolk. European Prospective Investigation into Cancer and Nutrition. AB - Previous studies have reported an association between plasma vitamin C levels and respiratory function, but have not examined the role of vitamin C as an effect modifier of the relation between cigarette smoking and obstructive airways disease (OAD). This question was investigated in a cross-sectional analysis of a population-based study of 3,714 males and 4,256 females aged 45-74 yrs. Undiagnosed OAD was defined as a forced expiratory volume in one second (FEV1) of <80% of the predicted value and FEV1/forced vital capacity of <70% without self reported OAD. An increase of 20 micromol x L(-1) (or 1 SD) in plasma vitamin C concentration was associated with a 13% reduction in the risk of having OAD (adjusted odds ratio (OR) (95% confidence interval) 0.87 (0.77-0.98)). The OR for current smokers relative to never smokers within the lowest quintile of plasma vitamin C concentration was 5.93 (3.03-11.61). The risk in the upper four quintiles was 2.84 (1.92-4.19). Within the lowest quintile of plasma vitamin C concentration, the risk in former smokers was strongly related to the time since quitting (interaction significant, p=0.001). These findings support a protective role for vitamin C against the risk of obstructive airways disease and support the hypothesis that vitamin C may be an effect modifier for the adverse effects of smoking on the risk of obstructive airways disease. PMID- 11028651 TI - Working in swine-confinement buildings causes an accelerated decline in FEV1: a 7 yr follow-up of Danish farmers. AB - Work in swine confinement units causes exposure to high levels of organic dust and is associated with a high prevalence of work-related respiratory symptoms and probably with accelerated decline in forced expiratory volume in one second (FEV1). A 7-yr follow-up on FEV1, forced vital capacity (FVC), bronchial reactivity, and respiratory symptoms was performed on 181 Danish farmers. The participation rate was 76.3% and nonparticipants had more symptoms, were more likely to be current-smokers and had lower lung function in the first survey than participants in both surveys. Farmers who worked exclusively with pigs in the follow-up had an accelerated decline in FEV1 but not in FVC compared with dairy farmers, where the observed decline in FEV1 was close to the expected. For a nonsmoking pig farmer compared to a nonsmoking dairy farmer the mean additional decline in FEV1 was 17 mL x yr(-1) (53.0 mL x yr(-1) versus 36.1 mL x yr(-1)). The authors conclude that working in swine confinement units causes an accelerated decline in forced expiratory volume in one second but not in forced vital capacity. The mean decline is approximately 0.5 L during a working life and some farmers will develop clinically significant airway obstruction due to work in swine confinement units. PMID- 11028652 TI - Exposure to antidepressants and the risk of cryptogenic fibrosing alveolitis: a case-control study. AB - The explanations for the emergence of cryptogenic fibrosing alveolitis as a new clinical entity during the second half of the 20th century are unclear. The authors have previously reported evidence of an increased risk of cryptogenic fibrosing alveolitis in relation to the use of antidepressant drugs. The authors have now tested this hypothesis a priori in an analysis of computerized general practice records for 890 cases of cryptogenic fibrosing alveolitis and 5,884 matched controls drawn from the UK General Practice Research Database. Exposure to antidepressants at the time of diagnosis was increased in cases compared to controls (odds ratio (OR) 1.52, 95% confidence interval (95% CI) 1.24-1.86), and this increase remained if the analysis was restricted to exposures 4 yrs prior to diagnosis (OR 1.50, 95% CI 0.98-2.30). However this increased prescribing was not specific to any particular class of antidepressant or individual drug, and there was no evidence of a dose-response relationship between exposure to amitriptyline (the most commonly prescribed antidepressant) and disease. The presented data do not allow any firm conclusion to be made as to whether there is a causal relationship between antidepressant exposure and cryptogenic fibrosing alveolitis, but it seems unlikely that exposure to tricyclic antidepressants shortly before diagnosis is a strong risk factor for cryptogenic fibrosing alveolitis. PMID- 11028653 TI - Association of Clara cell 10-kDa protein, spontaneous regression and sarcoidosis. AB - Sarcoidosis is a systemic granulomatous disorder with a high rate of spontaneous regression. Clara cell 10-kDa protein (CC10), the predominant product of nonciliated bronchiolar epithelial cells, is a potent immunoregulatory and anti inflammatory agent. CC10 levels were measured in sera and bronchoalveolar lavage (BAL) fluids from 31 sarcoidosis patients (nine progressive disease and 22 regressive disease) and their relevance to spontaneous regression investigated. The inhibitory effects of recombinant CC10 on interferon gamma (IFN-gamma) production were examined using lipopolysaccharide (LPS)-stimulated sarcoid BAL fluid cells, and the blocking effects of monoclonal antibody TY-5, directed against CC10, on CC10 function were also tested. Serum and BAL fluid CC10 levels in the regressive disease group were significantly higher than those in the progressive disease group (serum, p<0.05; BAL fluid, p<0.005) and healthy subjects (serum, p<0.0001; BAL fluid, p<0.005). CC10 inhibited, in part, IFN gamma production from LPS-stimulated sarcoid BAL fluid cells (CC10 inhibition: 1,000 ng x mL(-1), 30%; 100 ng x mL(-1), 14%). TY-5 restored IFN-gamma production by blocking CC10 function. Sarcoidosis patients with regressive disease showed increased Clara cell 10-kDa protein levels in their sera and bronchoalveolar lavage fluids. Clara cell 10-kDa protein may be a regulator of the inflammatory process in sarcoidosis. PMID- 11028654 TI - Comparison of asthma prevalence in the ISAAC and the ECRHS. ISAAC Steering Committee and the European Community Respiratory Health Survey. International Study of Asthma and Allergies in Childhood. AB - International and regional prevalence comparisons are required to test and generate hypotheses regarding the causes of increasing asthma prevalence in various age groups worldwide. The International Study of Asthma and Allergies in Childhood (ISAAC) is the first such study in children and the European Community Respiratory Health Survey (ECRHS) is the first such study in adults. Therefore, a comparison of the findings of these two surveys was conducted, for the 17 countries in which both surveys were undertaken. There was a strong correlation between the ISAAC and ECRHS prevalence data, with 64% of the variation at the country level, and 74% of the variation at the centre level, in the prevalence of "wheeze in the last 12 months" in the ECRHS phase I data being explained by the variation in the ISAAC phase I data. There was also generally good agreement in the international patterns observed in the two surveys for self-reported asthma (74% of country level and 36% of centre level variation explained), self-reported asthma before age 14 yrs (64 and 26%), hay fever (61 and 73%) and eczema (41 and 50%). Thus although there were differences in the absolute levels of prevalence observed in the two surveys, there is good overall agreement between the International Study of Asthma and Allergies in Childhood and European Community Respiratory Health Survey study findings with regard to international prevalence patterns. These findings, therefore, add support to the validity of the two studies, which provide a new picture of global patterns of asthma prevalence from child- to adulthood, and identify some of the key phenomena which future research must address. PMID- 11028655 TI - Hypomagnesaemia in chronic, stable asthmatics: prevalence, correlation with severity and hospitalization. AB - Magnesium (Mg) deficiency is associated with airway hyperreactivity, wheeze and impairment of lung function. The prevalence of hypomagnesaemia, and its effect on asthma is unclear. Therefore, this prospective study was undertaken to evaluate the prevalence of low serum Mg level, to correlate whether hypomagnesaemia (serum Mg level <0.74 mmol x L(-1)) is associated with severity of asthma and increased frequency of hospitalizations in chronic, stable asthmatics. Ninety-three chronic stable asthmatics, who were on regular follow-up in an asthma clinic, were studied. On attendance to the asthma clinic, each subject was interviewed and clinically evaluated. The serum Mg level (normal 0.74-1.2 mmol x L(-1)), as well as the severity of asthma was measured. The medications used were recorded. The total number of hospitalizations due to asthma exacerbation was scored for the last three months that preceded the date of measuring the serum Mg level. Twenty five patients (27%) had a low Mg level with a mean+/-SD of 0.69+/-0.04 mmol x L( 1), while 68/93 (73%) had a normal Mg level (0.82+/-0.08 mmol x L(-1)). The number of hospitalizations in asthmatics with low Mg levels was 10/25 (40%), compared to 8/68 (12%) (p<0.01) in asthmatics with normal Mg levels. Multiple logistic regression analysis showed that severe asthma was the only factor associated significantly with hypomagnesaemia (p=0.04). No effect was observed for inhaled beta-agonist, inhaled steroid or theophylline therapy on serum Mg level. To conclude, hypomagnesaemia is common in chronic asthmatics. Chronic asthmatics with low Mg tend to have more hospitalizations than chronic asthmatics with normal Mg. Hypomagnesaemia was also associated with more severe asthma. PMID- 11028656 TI - Natural history of aspirin-induced asthma. AIANE Investigators. European Network on Aspirin-Induced Asthma. AB - There is a subset of patients with bronchial asthma who are susceptible to disease exacerbation upon receiving aspirin and other nonsteroidal anti inflammatory drugs. This is a clinical syndrome, called aspirin-induced asthma (AIA), associated with alterations in arachidonate metabolism and cysteinyl leukotriene overproduction. The natural history and clinical characteristics of this type of asthma were studied. Sixteen clinical centres in 10 European countries provided standardized information to the specially developed patient oriented database regarding: medical history, physical examination, diagnosis, and treatment. Diagnosis of AIA was based on a typical history, confirmed by positive aspirin provocation tests, carried out in 91% of the patients. A total of 500 patients were enrolled in the study. AIA developed according to a pattern, characterized by a sequence of symptoms. First, persistent rhinitis, appearing at a mean age of 29.7+/-12.5 yrs, then asthma, aspirin intolerance and nasal polyposis appear. The clinical presentation in different European countries was remarkably similar. In females, who outnumbered males by 2.3:1, the onset of symptoms occurred significantly earlier and the disease was more progressive and severe than in males. Atopy, present in approximately a third of patients, led to earlier manifestation of rhinitis and asthma, but not of aspirin intolerance or nasal polyposis. A family history of aspirin intolerance, recorded in 6% of patients, had a less evident effect on the course of the disease than sex or atopy. Fifty one per cent of patients, in addition to inhaled steroids, required chronic systemic corticosteroid therapy at a mean dose of 8 mg prednisone x day( 1). Surprisingly, 15% of patients were unaware of intolerance to aspirin and learnt about it only after having provocation tests performed. All over Europe, aspirin-induced asthma develops in a similar characteristic way. Its course is influenced by sex and the presence of atopy. In half of the patients, asthma is severe, and steroid-dependent. The uniform natural history of aspirin-induced asthma might suggest a common underlying principle. PMID- 11028657 TI - Inhibition of nitric oxide synthase by nasal decongestants. AB - The nasal decongestants oxymetazoline and xylometazoline are frequently used in the topical treatment of rhinitis and sinusitis. As nitric oxide (NO) is thought to play a role in inflammation of the upper respiratory tract, the aim of this study was to examine the in vitro effects of these compounds on the activity and the expression of NO producing enzymes, including the inducible form of NO synthase (iNOS) and the constitutive isoform of NO synthase (cNOS). Experiments concerning the effects of both compounds on enzymatic activity and enzyme induction of iNOS were performed in a lipopolysaccharide (LPS) induced rat alveolar macrophage cell line (NR8383) using the Griess assay and the 3H citrulline assay respectively. The effects on cNOS were examined in fresh rat synaptosomes using the 3H-citrulline assay. The direct scavenging properties of both compounds were investigated using a amperometric NO sensor. Oxymetazoline and xylometazoline were shown to have a dose dependent inhibitory effect on total iNOS activity indicated by nitrite/nitrate formation in the Griess assay. This effect was found to be due to an inhibition of induction of the enzyme rather than inhibition of the enzyme activity, as was investigated in two separate experiments using the 3H-citrulline assay. Inhibition of cNOS was moderate and in the same order of magnitude as the inhibition of enzymatic iNOS activity. Direct scavenging of NO could not be detected. As constitutive nitric oxide synthase activity is thought to serve beneficial physiological functions, and exaggerated inducible nitric oxide synthase activity may cause exacerbation of the inflammatory process, pharmacological treatment influencing the nitric oxide generating system should focus on inhibition of inducible nitric oxide synthase alone. The specific characteristics of these decongestants in vitro suggests suitability for this application and may indicate an additional beneficial effect in the treatment of upper respiratory tract inflammation. PMID- 11028658 TI - Increased vascular expression of iNOS at day but not at night in asthmatic subjects with increased nocturnal airway obstruction. AB - Nitric oxide production by endothelial cells may have important consequences for the development of airway inflammation as well as for airway obstruction. The present study investigated whether the expression of vascular inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in human bronchi differs between asthmatic and healthy subjects, and whether it shows a circadian rhythm, especially in subjects with increased nocturnal airway obstruction. Bronchial biopsy samples were taken at 16:00 and 04:00 h from 13 healthy and 25 asthmatic subjects, 18-45 yrs. Biopsy samples were snap-frozen and double-immunostained for iNOS and eNOS in combination with a common vascular antigen (CD31). The degree of immunopositivity was expressed as a percentage of CD31-positive vessels encountered in complete biopsy sections. Asthmatic subjects showed greater iNOS expression than healthy controls: 23+/-15 versus 7+/-17% (mean+/-SD) at 16:00 h (p<0.001) and 19+/-15 versus 8+/-11% at 04:00 h (p<0.05). Asthmatic subjects with a fall in forced expiratory volume in one second of >10% of the predicted value between 16:00 and 04:00 h showed greater iNOS expression at 16:00 than at 04:00 h: 32+/-16 versus 20+/-13% (p<0.05). eNOS expression did not differ between healthy controls and asthmatic patients, nor did it differ between 16:00 and 04:00 h. It is suggested that asthmatic subjects with increased nocturnal airway obstruction demonstrate increased activation of inducible nitric oxide synthase during the day. The resulting nitric oxide production might protect against airway obstruction during the day. However, at night, nitric oxide production is probably insufficient to counterbalance the bronchoconstricting forces. PMID- 11028660 TI - Classical protein kinase C and its hypoxic stimulus-induced translocation in the cat and rat carotid body. AB - The presence, subcellular distribution, species specificity and possible hypoxic stimulus-induced translocation of classical protein kinase C (cPKC) isozymes were examined in the carotid body. Carotid bodies were dissected from cats exposed in vivo to normoxic or acute hypoxic conditions and from normoxic rats. For immunohistochemistry isoform-specific monoclonal antisera to PKCalpha, PKCbetaI, PKCbetaII and PKCgamma were used. The immunoreactivity was visualized by fluorescein isothiocyanate (FITC) labelling. FITC/Texas red double-labelled specimens for the cPKC isozymes/tyrosine hydroxylase were used to demonstrate the chemoreceptor cell localization of cPKC isozymes. The immunofluorescence was detected using laser scanning confocal image technology. The results showed expression of the PKCalpha and PKCgamma but not PKCbeta isoforms in the cytoplasm of carotid body chemoreceptor cells. The double labelling provided evidence for the chemoreceptor cell localization of the cPKC isoforms detected. The immunostaining was most intense in the periphery of the perikarya, the nuclear envelope and, occasionally, the nucleoplasm. No major differences were found in the immunolocalization of PKCalpha and PKCgamma under normoxic and hypoxic conditions or between species. However, the immunoreactivity tended to accumulate more in the peripheral cytoplasm and away from the nucleus in the hypoxic chemoreceptor cell. This study demonstrates the presence of classical protein kinase C enzymes in chemoreceptor cells. The intensity of the immunoreactivity may suggest a role for the classical protein kinase C signalling pathway in shaping the hypoxic response at the carotid body. However, this study failed to provide firm evidence of this. PMID- 11028659 TI - Bradykinin stimulates IL-6 and IL-8 production by human lung fibroblasts through ERK- and p38 MAPK-dependent mechanisms. AB - Bradykinin (BK) is a major kinin with well-documented pharmacological properties including vascular leakage and induction of a variety of cytokines. However, the intracellular signalling mechanisms by which BK induced proinflammatory cytokine production have not been fully elucidated. This study investigated the role of the extracellular signal-regulated protein kinase 1/2 (ERK 1/2) and p38 mitogen activated protein kinase (p38 MAPK) in the BK-induced interleukin (IL)-6 and IL-8 production by human lung fibroblasts. Lung fibroblasts were stimulated with BK in the presence or in the absence of PD98059, a specific MAPK/ERK kinase-1 inhibitor, or SB203580, a specific p38 MAPK inhibitor, and IL-6 or IL-8 production and their gene expression was examined. BK-induced ERK 1/2 or p38 MAPK phosphorylation was also analysed by Western blot analysis. BK at nanomolar concentrations stimulated lung fibroblasts to produce IL-6 and IL-8 along with increased ERK 1/2 and p38 MAPK phosphorylation. BK-induced IL-6 and IL-8 synthesis was inhibited by a B2-type BK receptor antagonist. Furthermore, PD98059 or SB203580 significantly suppressed BK-induced IL-6 and IL-8 production and their gene expression. These results indicate that bradykinin-induced interleukin 6 and interleukin-8 production are at least partly mediated through the extracellular signal-related protein kinase 1/2 and p38 mitogen-activated protein kinase pathway-dependent activation in human lung fibroblasts, and suggest that bradykinin appears to be involved in the inflammatory reaction leading to acute lung injury through stimulating interleukin-6 and interleukin-8 production by lung fibroblasts. PMID- 11028661 TI - Allergic bronchopulmonary aspergillosis in cystic fibrosis. A European epidemiological study. Epidemiologic Registry of Cystic Fibrosis. AB - Allergic bronchopulmonary aspergillosis (ABPA) is a disease resulting from a hypersensitivity response to Aspergillus fumigatus, although the pathogenesis of the disease is unknown and its prevalence in cystic fibrosis (CF) is still poorly defined. Data from the Epidemiologic Registry of Cystic Fibrosis (ERCF) on 12,447 CF patients gathered from 224 CF centres in nine European countries were analysed. The ERCF definition of ABPA diagnosis is a positive skin test and serum precipitins to A. fumigatus, together with serum immunoglobulin (Ig)E levels >1,000 U x mL(-1) and additional clinical or laboratory parameters. The overall prevalence of ABPA in the ERCF population was 7.8% (range: 2.1% in Sweden to 13.6% in Belgium). Prevalence was low <6 yrs of age but was almost constant approximately 10% thereafter. No sex differences were observed. ABPA affected 8.0% of patients with a deltaF508/deltaF508 genotype and 5-6% with deltaF508/G551D, deltaF508/G542X and deltaF508/N1303K genotypes. ABPA patients presented a lower forced expiratory volume in one second (FEV1) than those without ABPA at any age and the prevalence ranged from 6.6% in patients with FEV1 > or =20-12.9% in those with FEV1 <40%. ABPA was associated with higher rates of microbial colonization, pneumothorax and massive haemoptysis, and with higher IgG serum levels and poorer nutritional status. A mixed model regression analysis of lung function showed that FEVI decline during the follow-up period was not substantially different in ABPA patients compared with non-ABPA patients for any subgroups based on age or disease severity at enrollment. To conclude, allergic bronchopulmonary aspergillosis is a frequent complication in cystic fibrosis patients, particularly after the age of 6 yrs, and it is generally associated with a poorer clinical condition. However, any clear independent influence of allergic bronchopulmonary aspergillosis on the rate of lung function decline in the short term was not shown. PMID- 11028662 TI - Observer variability of lung function measurements in 2-6-yr-old children. AB - The aim of this study was to assess the within-observer and between-observer variability of lung function measurements in children aged 2-6 yrs. Two observers examined 22 asthmatic children independently according to a predefined protocol. Each observer obtained duplicate measurements of respiratory resistance by the interrupter technique (Rint), respiratory resistance (Rrs,5) and reactance (Xrs,5) at 5 Hz by the impulse oscillation technique and the specific airway resistance (sRaw) by whole body plethysmography. The within-subject SD (SDw) was not significantly different in the two observers. The ratio SDw between observers/mean SDw within observers was 0.94, 1.25, 1.35 and 2.86 for Xrs,5, Rrs,5, sRaw and Rint, respectively, indicating greater between-observer variability of the latter. The systematic difference between observers assessed by the difference between observer means (expressed as a percentage of their mean value) was 11, 7, 6 and 2% for Xrs,5, sRaw, Rrs,5 and Rint, respectively. These differences were statistically significant, except that for Rint. In conclusion, specific airway resistance, impulse oscillation technique and respiratory resistance assessed by the interrupter technique measurements in young children are subject to influence by the observer, and the random variability between observers appears to be particularly great for respiratory resistance assessed by the interrupter technique. The authors suggest that the between-observer variability should be investigated when evaluating novel methods for testing lung function. PMID- 11028663 TI - Outcome of paediatric domiciliary mask ventilation in neuromuscular and skeletal disease. AB - Noninvasive positive pressure ventilation delivered by nasal mask or facemask has been used widely in the last decade to manage chronic ventilatory failure in adults with neuromuscular and chest wall disease. However, it has been thought that paediatric patients would not be able to tolerate masks, and previous anecdotal reports on the paediatric application of mask ventilation have not assessed the effects on nocturnal and arterial blood gas control. Domiciliary mask ventilation has been used in 40 children with ventilatory insufficiency due to congenital neuromuscular and skeletal disease aged 9 months-16 yrs. Eighteen patients had symptomatic nocturnal hypoventilation, 17 had diurnal ventilatory failure, three were referred for weaning and two had frequent chest infections associated with sleep-disordered breathing. Thirty eight of the 40 patients tolerated mask ventilatory support long-term. Diurnal mean+/-SD oxygen tension in arterial blood (Pa,O2) increased from 8.5+/-1.8-10.9+/-1.7 kPa (p<0.001) and mean carbon dioxide tension in arterial blood (Pa,CO2) fell from 7.0+/-1.6-5.9+/-0.8 kPa (p=0.01) following initiation of ventilatory support. Mean and minimum nocturnal Pa,O2 and peak transcutaneous carbon dioxide tension (Ptc,CO2) (n=21) improved significantly. Mask ventilation can be used successfully in young children and reverses ventilatory insufficiency due to congenital neuromuscular and skeletal disease. PMID- 11028664 TI - Effect of preoperative hyperinflation on static lung volumes after lung transplantation. AB - It is still not known whether persistent increases in functional residual capacity (FRC) and residual volume (RV) after lung transplantation are due to preexisting hyperinflation. Therefore, the aim of this study was to determine the effects of chronic lung hyperinflation on static lung volumes after heart/lung (HLT) and bilateral lung transplantation (BLT). Static lung volumes were measured in 33 patients before and at 6 month intervals for up to 3 yrs after HLT (n=25) or BLT (n=8). The preoperative diagnosis was cystic fibrosis in 25 patients and other chronic hyperinflated lung diseases in eight patients. After surgery, total lung capacity returned to predicted normal values but FRC and RV remained greater than expected for either the recipient or the donor. At 1 yr after surgery, mean+/-SD FRC and RV were 130+/-18% and 151+/-34% of the predicted values for the recipient (p<0.001), and these figures did not change significantly over time. Similar abnormalities were found in patients with and without cystic fibrosis. After transplantation for lung diseases producing chronic hyperinflation, there is a persistent increase in functional residual capacity and residual volume. This alteration is present in patients operated on for diseases developed in both childhood and adulthood and is not recovered over time. It may be due to irreversible changes in the structure of the ribcage. PMID- 11028665 TI - Borg scores before and after challenge with adenosine 5'-monophosphate and methacholine in subjects with COPD and asthma. AB - Dyspnoea differs between subjects with chronic obstructive pulmonary disease (COPD) and asthma, partly because the underlying mechanisms for bronchoconstriction differ. This study investigated the possible role of inflammation and the contribution of clinical variables on dyspnoea in subjects with COPD and asthma. Forty-eight smoking subjects with COPD and 21 nonsmoking subjects with asthma, were challenged with adenosine 5'-monophosphate (AMP) and methacholine. The Borg score was assessed before and after each challenge. Mean increases in Borg score (per percentage decrease in baseline forced expiratory volume in one second (FEV1)) were significantly smaller in COPD than in asthma (p<0.01), values being 0.055 and 0.045 in COPD and 0.122 and 0.093 in asthma respectively. This difference was largely due to the fact that one-third of the subjects with COPD did not increase their Borg score during bronchoconstriction. The increase in Borg tended to be larger during AMP than during methacholine challenge, both in asthma and COPD. Changes in Borg scores were explained by age in COPD and by the Borg score before AMP challenge in asthma. The authors conclude that perception of dyspnoea during adenosine 5'-monophosphate and methacholine induced bronchoconstriction is lower in chronic obstructive pulmonary disease than in asthma and that age contributes to this difference. As adenosine 5'-monophosphate is regarded as an indirect marker of airway inflammation, the results suggest that inflammation is not important because both groups showed similar responses on such provocations. PMID- 11028666 TI - Nasal proportional assist ventilation unloads the inspiratory muscles of stable patients with hypercapnia due to COPD. AB - This study was undertaken to assess the physiological effects of proportional assist ventilation (PAV), administered noninvasively through a nose mask, on ventilatory pattern, arterial blood gases, lung mechanics, and inspiratory muscle effort in stable, hypercapnic patients with chronic obstructive pulmonary disease. In 15 patients, PAV was set by adjusting volume assist (VA) and flow assist (FA) according to the "run-away" technique and the patient's comfort respectively. The level of support was fixed at 80% of the total possible assistance and averaged 13.9+/-4.1 cmH2O x L(-1) and 4.1+/-1.3 cmH2O x L(-1) x s for VA and FA, respectively. Continuous positive airway pressure (CPAP) was established at 2 cmH2O and then increased to 5 cmH2O. Physiological measurements were made during spontaneous breathing (SB), after more than 40 min of PAV, and 20 min after the rise in CPAP. On average, PAV improved ventilation (10.3+/-2.1 to 12.5+/-2.0 L x m(-1)), tidal volume (0.60+/-0.11 to 0.76+/-0.24 L), arterial oxygen tension and arterial carbon dioxide pressure (from 6.7+/-0.7 to 7.1+/-0.9 and from 7.6+/-1.0 to 7.2+/-1.2 kPa, respectively). During SB, pulmonary resistance and dynamic lung elastance averaged 15.0+/-7.6 cmH2O x L(-1) s and 15.8+/-8.0 cmH2O x L(-1), respectively. Assuming a normal chest wall elastance (5 cmH2O x L(-1)), VA and FA relieved respectively approximately 70% of the elastic and 30% of the resistive burden, with PAV set with the procedure of this study. The overall magnitude of the patients' inspiratory effort, measured by means of the oesophageal and diaphragmatic pressure time product in 10 patients was significantly reduced by PAV, on average, 328+/-122 to 226+/-118 (-31%) and 361+/ 119 to 254+/-126 (-30%) cmH2O x min(-1), respectively. In 10 patients the electrical activity of the diaphragm (Edi) was also reduced by PAV to approximately 70%, on average, of the SB activity. The rise of CPAP 25 cmH2O did not cause any further significant change in the physiological variables. In all instances there was a good patient-ventilator interaction, the ventilatory breath never entering into the patient's neural expiratory time. These data show that nasal proportional assist ventilation can provide physiological benefits to the stable hypercapnic chronic obstructive pulmonary disease patients. In fact, proportional assist ventilation, which was well tolerated by all patients, unloaded the inspiratory muscles and improved arterial blood gases. Further studies can clarify whether these beneficial physiological effects of nasal proportional assist ventilation can bear profitable consequences in the overall clinical management of chronic obstructive pulmonary disease patients with chronic carbon dioxide retention. PMID- 11028667 TI - NO chemiluminescence in exhaled air: interference of compounds from endogenous or exogenous sources. AB - Nitric oxide determination in exhaled air using chemiluminescence analysers is increasingly used, but may be affected by various other components of the air sample. The influence of several compounds originating from endogenous or exogenous sources on NO readings has been studied. Defined amounts of water vapour, carbon dioxide, acetone, heptane, acetonitrile, oxygen, nitrous oxide and enflurane were added to air samples with NO concentrations 0-250 parts per billion. Marked and significant decreases in NO readings, which strongly depend on the concentration of the respective interfering compound, were found for water vapour (0.155% per 1% relative humidity), carbon dioxide (1.97% per 1% CO2 volume/volume (v/v)) and nitrous oxide (0.608% per 1% v/v N2O). While acetone in concentrations up to 8.5% v/v had no measurable effect on NO readings, heptane and acetonitrile led to marked decreases. Oxygen in concentrations of up to 95% (v/v) had no effect on NO determination. NO readings were markedly decreased by >10% per 1% (v/v) of the anaesthetic enflurane. However, due to large variations in NO values, these decreases were not statistically significant. Furthermore, enflurane reacted with the molybdenum converter of the NO(x) analyser in use, resulting in major damage to the instrument. Eliminating, or at least considering, interferences by compounds present in exhaled air is an urgent prerequisite for reliable and precise nitric oxide determination. PMID- 11028668 TI - Enhanced peripheral leukocyte leukotriene production and bronchial hyperresponsiveness in asthmatics. AB - Leukotrienes (LTs) are pro-inflammatory mediators that contribute to the pathophysiological features of asthma. The relationship between the amounts of LTB4 and LTC4 produced by the leukocytes of asthmatic patients on the one hand and immunoglobulin E (IgE)-mediated allergy, asthma exacerbations and bronchial hyperresponsiveness was studied. Leukocytes were obtained from peripheral blood drawn from 29 atopic and 27 nonatopic asthmatics during exacerbations and clinically controlled periods, as well as from 20 control individuals. The leukocytes were stimulated with calcium ionophore A23187 to induce LTB4 and LTC4 production. Allergy was assessed by means of specific serum IgE or by positive skin tests, whereas bronchial hyperresponsiveness was measured by methacholine challenge. The leukocytes of the asthmatics generated significantly more LTB4 (p<0.05) and LTC4 (p<0.01) than those of controls. The leukocytes of patients with atopic asthma generated significantly more LTC4 than those of patients with nonatopic asthma (p<0.01). Significantly more LTC4 was produced by leukocytes obtained during exacerbations, than by those obtained during clinically controlled periods (p<0.01). In addition, there was a significant correlation between LTB4 generation by leukocytes and the degree of bronchial hyperresponsiveness to methacholine (r=-0.792, p<0.0001). These results suggest that leukotriene C4 production by leukocytes is associated with immunoglobulin E mediated allergy and asthma exacerbations, and further that generation of leukotriene B4 is closely related to bronchial hyperresponsiveness in patients with asthma. PMID- 11028669 TI - Interstitial lung disease in children: a multicentre survey on diagnostic approach. AB - Chronic interstitial lung disease (ILD) is a rare disorder in the paediatric age group, with a poor prognosis. The diagnostic approach to ILD is based on more or less invasive methods. This study was implemented to verify which methods are the most often used in children. Questionnaires (333) were sent to members of the European Respiratory Society Paediatric Assembly belonging to 187 European and non-European centres. Questions concerned the use of noninvasive diagnostic methods, e.g. history taking, physical examination, routine laboratory tests, respiratory function tests and radiology (chest radiography, high-resolution computed tomography (HRCT)), and the use of invasive techniques such as bronchoalveolar lavage (BAL), transbronchial biopsy (TBB), open lung biopsy (OLB), video-assisted thoracoscopic biopsy (VAT) and HRCT with fine-needle aspiration biopsy (FNAB). Thirty eight centres returned the questionnaires and 131 children with ILD were studied. A diagnosis of ILD was achieved in five (3.8%) patients using noninvasive techniques alone. Using the various biopsy methods, histological assessment was performed on a total of 98 (74.8%) children. The most frequently used invasive technique both alone and in combination was BAL (83, 63.3%), followed by OLB (64, 48.8%), TBB (26, 19.8%) and VAT (11, 8.4%); FNAB was used in one patient. In conclusion a diagnosis of interstitial lung disease was reached on the basis of aetiological and/or histological findings in 117 (89%) of the 131 patients studied. PMID- 11028670 TI - Indirect bronchial hyperresponsiveness in asthma: mechanisms, pharmacology and implications for clinical research. AB - Bronchial hyperresponsiveness (BHR), an abnormal increase in airflow limitation following the exposure to a stimulus, is an important pathophysiological characteristic of bronchial asthma. Because of heterogeneity of the airway response to different stimuli, the latter have been divided into direct and indirect stimuli. Direct stimuli cause airflow limitation by a direct action on the effector cells involved in the airflow limitation, while indirect stimuli exert their action essentially on inflammatory and neuronal cells that act as an intermediary between the stimulus and the effector cells. This manuscript reviews the clinical and experimental studies on the mechanisms involved in indirect BHR in patients with asthma. Pharmacological stimuli (adenosine, tachykinins, bradykinin, sodium metabisulphite/sulphur dioxide, and propranolol) as well as physical stimuli (exercise, nonisotonic aerosols, and isocapnic hyperventilation) are discussed. The results of the different direct and indirect bronchial challenge tests are only weakly correlated and are therefore not mutually interchangeable. Limited available data (studies on the effects of allergen avoidance and inhaled corticosteroids) suggest that indirectly acting bronchial stimuli (especially adenosine) might better reflect the degree of airway inflammation than directly acting stimuli. It remains to be established whether monitoring of indirect BHR as a surrogate marker of inflammation (in addition to symptoms and lung function) is of clinical relevance to the long-term management of asthmatic patients. This seems to be the case for the direct stimulus methacholine. More work needs to be performed to find out whether, indirect stimuli are more suitable in asthma monitoring than direct ones. Recommendations on the application of indirect challenges in clinical practice and research will shortly be available from the European Respiratory Society Task Force. PMID- 11028671 TI - Oxidative stress and regulation of glutathione in lung inflammation. AB - Inflammatory lung diseases are characterized by chronic inflammation and oxidant/antioxidant imbalance, a major cause of cell damage. The development of an oxidant/antioxidant imbalance in lung inflammation may activate redox sensitive transcription factors such as nuclear factor-KB, and activator protein 1 (AP-1), which regulate the genes for pro-inflammatory mediators and protective antioxidant genes. Glutathione (GSH), a ubiquitous tripeptide thiol, is a vital intra- and extracellular protective antioxidant against oxidative/nitrosative stresses, which plays a key role in the control of pro-inflammatory processes in the lungs. Recent findings have suggested that GSH is important in immune modulation, remodelling of the extracellular matrix, apoptosis and mitochondrial respiration. The rate-limiting enzyme in GSH synthesis is gamma-glutamylcysteine synthetase (gamma-GCS). The human gamma-GCS heavy and light subunits are regulated by AP-1 and antioxidant response elements and are modulated by oxidants, phenolic antioxidants, growth factors, and inflammatory and anti inflammatory agents in lung cells. Alterations in alveolar and lung GSH metabolism are widely recognized as a central feature of many inflammatory lung diseases such as idiopathic pulmonary fibrosis, acute respiratory distress syndrome, cystic fibrosis and asthma. The imbalance and/or genetic variation in antioxidant gamma-GCS and pro-inflammatory versus antioxidant genes in response to oxidative stress and inflammation in some individuals may render them more susceptible to lung inflammation. Knowledge of the mechanisms of GSH regulation and balance between the release and expression of pro- and anti-inflammatory mediators could lead to the development of novel therapies based on the pharmacological manipulation of the production as well as gene transfer of this important antioxidant in lung inflammation and injury. This review describes the redox control and involvement of nuclear factor-kappaB and activator protein-1 in the regulation of cellular glutathione and gamma-glutamylcysteine synthetase under conditions of oxidative stress and inflammation, the role of glutathione in oxidant-mediated susceptibility/tolerance, gamma-glutamylcysteine synthetase genetic susceptibility and the potential therapeutic role of glutathione and its precursors in protecting against lung oxidant stress, inflammation and injury. PMID- 11028672 TI - Modelling the production of nitric oxide within the human airways. AB - The measurement of exhaled nitric oxide (NO) is well established for monitoring of airway inflammation in bronchial asthma. It is known that the concentration of NO determined as steady state (plateau) value at a constant expiratory flow rate depends on the flow rate chosen. This suggests that the exhaled NO is released within the conducting airways, whereas alveolar NO levels are negligible. The processes involved can be described through a lung model comprising an alveolar compartment and an airway compartment thought of as a pipe. This concept has been formulated mathematically and the models proposed in the literature are essentially equivalent. NO plateau levels obtained at different flow rates allow the estimation of 1) an effective airway wall NO concentration that represents the driving force for NO release, 2) an airway diffusing capacity for NO which depends on factors impeding or facilitating NO transport, including an increase in NO-producing surface area. Clinical studies will have to assess whether the knowledge of these or related parameters offers a significant advantage over the determination of exhaled NO at a single flow rate. PMID- 11028673 TI - How the measurement of residual volume developed after Davy (1800). AB - H. Davy measured the residual volume of his own lungs in 1800, by inhaling a hydrogen mixture contained in a mercurial air holder. Using the same principle, Nestor Grehant determined the functional residual capacity, and the volume of the dead space, in 1864. Both used a forced breathing method, that was substituted by a prolonged dilution method by D.D. Van Slyke and C.A.L. Binger in 1923. It was in 1941 that G.R. Meneely and M.L. Kaltreider replaced hydrogen with helium. The open circuit nitrogen washout method was proposed by R.E. Darling, A. Cournand and D.W. Richards in 1940, and the body plethysmograph by A.B. DuBois et al. in 1956. So the three methods, still in common use today for measuring the static lung volumes, had been described by the mid-1950s. PMID- 11028674 TI - An elderly female with dyspnoea and anaemia. PMID- 11028675 TI - Severe asthma and ARDS triggered by acute short-term exposure to commonly used cleaning detergents. AB - This report presents a case of severe asthmatic attack and adult respiratory distress syndrome, which occurred in an asthmatic subject following acute short term exposure to commonly used cleaning detergents. Premorbid lung conditions, i.e. asthma and airway hyperresponsiveness, might have been risk factors for illness severity but not for persistent complaints. This case report indicates that nonindustrial exposures due to household product misuse, especially to mixing cleaning agents, may cause adult respiratory distress syndrome and not only irritant-induced asthma. PMID- 11028676 TI - Role of induced sputum in interstitial lung disease. PMID- 11028677 TI - Smoking and asthma. PMID- 11028678 TI - Nocturnal ventilation in neuromuscular diseases. PMID- 11028679 TI - Low asthma prevalence in Turkish children. PMID- 11028681 TI - Brain lipoprotein metabolism and its relation to neurodegenerative disease. AB - Lipoproteins are macromolecular complexes composed of lipids and proteins. The role of these complexes is to provide cells of the organism with lipids to be used as a source of energy, building blocks for biomembrane synthesis, and lipophilic molecules (e.g., steroid hormones and vitamin E) for other physiological purposes, such as cell signaling and antioxidative mechanisms. Lipoproteins also promote the cellular efflux of cholesterol for its disposal into bile. Thus, lipoproteins play an important role in the maintenance of lipid homeostasis throughout the organism. Accordingly, lipoprotein particles have been found circulating in blood, lymph, and interstitial fluid. Despite the existence of the blood-brain barrier, lipoprotein particles have been shown to be also present in the cerebrospinal fluid (CSF). Although a portion of their protein components may filter through the barrier from the vascular compartment, experimental evidence indicates that these particles originate from the nervous tissue. The other protein components include apolipoproteins E, J, and D, and these have been shown to be synthesized by cells within the central nervous system (CNS). Furthermore, it was shown that lipoprotein particles can be isolated from the conditioned medium of astrocytic cultures. The differences in size, structure, and composition of in vitro assembled particles compared with those isolated from the CSF suggest that the particles are modified following their secretion in vivo. This is supported by observations that lipoprotein modifying enzymes and transfer proteins are also present within CNS tissue and CSF. The fate of CSF lipoproteins is unclear but is probably related to the turnover and clearance of lipids from the CNS or, alternatively, the particles may be recaptured and recycled back into the CNS tissue. The presence of several cell surface receptors for apoE-containing lipoproteins on ependymal cells, as well as on neurons and glial cells, supports this notion and suggests that the isolated brain possesses its own system to maintain local lipid homeostasis. This is further exemplified by the salvage and recycling of lipids shown to occur following a lesion in order to allow surviving neurons to sprout and reestablish lost synapses. Not much is currently known about lipoprotein metabolism in neurodegenerative diseases, but lipid alterations have been repeatedly reported in Alzheimer brains in which neuronal loss and deafferentation are major features. Although the mechanism underlying the link between the epsilon4 allele of the apolipoprotein E gene and Alzheimer's disease is presently unclear, it may well be postulated that it is related to disturbances in brain lipoprotein metabolism. PMID- 11028680 TI - A noradrenergic and serotonergic hypothesis of the linkage between epilepsy and affective disorders. AB - Noradrenergic and/or serotonergic deficits, as well as other abnormalities, may contribute to predisposition to some epilepsies and depressions. Evidence for this hypothesis stems from several sources. Epidemiological investigations are intriguing but incomplete. Pharmacological studies show that noradrenergic and/or serotonergic transmission are both anticonvulsant and antidepressant. Therapeutically pertinent investigations show that antidepressant drugs have anticonvulsant properties, whereas antiepileptic drugs are effective in the management of affective disorders. Additional investigations demonstrate that seizures, whether spontaneously occurring or therapeutically induced, protect against depression. Through studies of innate pathophysiology, noradrenergic and serotonergic deficits have been identified in individuals with depression and in animal models of epilepsy, as well as in some humans with epilepsy. Vagal nerve stimulation, a treatment already known to be effective in the epilepsies, is presently under investigation for effectiveness in affective disorder. New evidence suggests that vagal nerve stimulation exerts at least some of its therapeutic effects through its capacity to increase noradrenergic and serotonergic transmission. Finally, emerging evidence supports the concept that some genetic mammalian models of the human epilepsies exhibit analogous manifestations of depression. PMID- 11028682 TI - Microchip technology in drug delivery. AB - The realization that the therapeutic efficacy of certain drugs can be affected dramatically by the way in which they are delivered has created immense interest in controlled drug delivery systems. Much previous work in drug delivery focused on achieving sustained drug release rates over time, while a more recent trend is to make devices that allow the release rate to be varied over time. Advances in microfabrication technology have made an entirely new type of drug delivery device possible. Proof-of-principle experiments have shown that silicon microchips have the ability to store and release multiple chemicals on demand. Future integration of active control electronics, such as microprocessors, remote control units, or biosensors, could lead to the development of a 'pharmacy on a chip,' ie 'smart' microchip implants or tablets that release drugs into the body automatically when needed. PMID- 11028683 TI - Adrenergically stimulated fat utilization and ageing. AB - Ageing is associated with a diminished ability to use fat as a fuel during exercise. Also, middle-aged subjects have a blunted ability to mobilize fatty acids and to increase skeletal muscle fatty acid uptake and oxidation during intravenous beta-adrenergic stimulation, indicating that the sympathetic nervous system may play a role in the disturbed fat utilization. The blunted lipolytic response may be related to disturbances at the receptor level, eg a diminished number or agonist affinity of beta-adrenoceptors, or at the postreceptor level, eg a diminished activity of the hormone-sensitive lipase complex. As the rates of fatty acid availability are not limiting during exercise or beta-adrenergic stimulation in the elderly, the lowered skeletal muscle fat oxidation is probably related to an age-related decline in the capacity of skeletal muscle to oxidize fatty acids. Factors responsible for this decline may be a diminished content of oxidative enzymes, an increased glycolytic flux inhibiting fatty acid transport into the mitochondria, or a diminished (possibly beta-adrenergically-mediated) activation of fatty acid transport. It remains to be determined to what extent disturbances of fat metabolism may be related to the ageing process per se or whether they are secondary to age-related changes in body fat distribution and level of physical activity. Nevertheless, the impairments in sympathetically mediated lipolysis and fat oxidation may be of importance in the age-related increase in adiposity and insulin resistance and may thus be one of the links between ageing and increased prevalence of chronic diseases, such as obesity, type 2 diabetes mellitus, and cardiovascular disease. PMID- 11028684 TI - Arterial stiffness and the development of hypertension. AB - Evidence is accumulating in support of the plausible notion that lower arterial elasticity is associated with an increased risk of cardiovascular disease manifestations. Conventional indices of arterial stiffness are compared to a newer method (blood pressure-adjusted pulsatile arterial diameter change) in terms of blood pressure dependence. Impaired elasticity of larger arteries is an antecedent factor in the natural history of blood pressure elevation at the population level. Strong evidence supports the notion that lower arterial elasticity in the medium-sized arteries is associated with the development of hypertension. PMID- 11028685 TI - Angiotensin AT1 receptor over-expression in hypercholesterolaemia. AB - Angiotensin II mediates most of the biological effects of the renin-angiotensin system (RAS), such as vasoconstriction and cell proliferation, via stimulation of the angiotensin II type 1 (AT1) receptor. The AT1 receptor plays a central role in the pathogenesis of atherosclerosis and hypertension. In parallel, hypercholesterolaemia is a major risk factor for the development and progression of cardiovascular diseases. The underlying molecular events, however, are understood only partially. An important mechanism may be the interaction between hypercholesterolaemia and AT1 receptor expression in vascular tissue. Low-density lipoprotein (LDL) cholesterol leads to a profound increase in AT1 receptor expression in cultured vascular smooth muscle cells as well as in hypercholesterolaemic rabbits. This up-regulation is associated with an enhanced functional response upon stimulation with angiotensin II. Over-expression of the vascular AT1 receptor can also be observed in hypercholesterolaemic men and is prevented by treatment with 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors. These findings may explain why hypercholesterolaemia is frequently associated with hypertension and why blockade of the RAS attenuates the progression of atherosclerosis. PMID- 11028686 TI - Daycare attendance, asthma and atopy. AB - The objective of this article is to review studies that have examined the relation of daycare to asthma and atopy. In order to identify studies for inclusion, abstracts of all studies referenced in Medline from January 1966 to January 2000 and in BIBSYS were searched and extracted if they included 'asthma' or 'atopy' combined with words such as 'daycare', 'nursery' or 'kindergarten'. Eight studies fulfilled the criteria of inclusion. The outcomes were asthma, skin prick test (SPT) reactivity, a positive radioallergosorbent test (RAST), hay fever, and eczema. Daycare attendance was positively associated with asthma in five of six studies including asthma. In three of these studies there was no statistically significant association between daycare and asthma. Early start in daycare protected against later asthma in one study. There was a weak, but not a statistically significant positive relation between daycare and atopy in two of three studies when SPT reactivity was used as the outcome. In children of small families early start in daycare protected against atopy. The quality of the studies varies, and they are not directly comparable. The relation between daycare attendance and asthma and atopy is unclear, and further studies designed to answer this specific research question are needed. PMID- 11028687 TI - Puzzling associations between childhood infections and the later occurrence of asthma and atopy. AB - The current unfavourable trends in asthma and atopy prevalences have raised great concern and have challenged investigators to accelerate search for new risk factors for atopic diseases. The lack or scarcity of intense, systemic infections in early life has been postulated to increase susceptibility of becoming sensitized to otherwise harmless allergens in later life. This hygiene hypothesis is considered one of the most plausible explanations for the current trends in atopic diseases to date. There are data to suggest that measles, hepatitis A, and Mycobacterium tuberculosis infection in early life may prevent the subsequent development of atopic diseases. The hypothesis is based on the concept that certain viral and bacterial infections, which induce a strongly polarized T helper (Th)-1 type response and a long-lasting memory immunity, are in early life able to reverse or prevent the biased Th1/Th2 balance in individuals prone to atopy and asthma. Evidence for the ability of mycobacterial infections to alter the Th1/Th2 balance has also been obtained from murine models. In humans, the critical time period during which immunomodulation with long-lasting effects is considered most successful is within the first two years of life. Possibly also nonpathogenic residents of the intestinal mucosa are involved in the proper maturation of the immune system. The use of antibiotics has been shown to be positively associated with the development of asthma and atopy. The mechanisms underlying these associations remain largely unknown. PMID- 11028688 TI - Consequences of adolescent drug use on psychiatric disorders in early adulthood. AB - This article summarizes the existing literature on the relationship between adolescent drug use and abuse and the development of psychiatric disorders in adulthood. In recent years, there has been increased awareness of the co occurrence of drug abuse and psychiatric disorders in adolescence and young adulthood. Few longitudinal studies, however, have examined specifically the impact of earlier drug use and abuse on later psychiatric disorders. The literature suggests three possible models to explain the relation between drug use and abuse and psychiatric disorders. According to the first model, adolescent psychiatric disorders precede drug use and abuse. A second model postulates that psychiatric disorders and drug use are correlated because they share one or more common aetiological factor(s). The third model posits that drug use and abuse predict or precede certain psychiatric disorders. We present data from a recent longitudinal study to support this latter model. As drug use and abuse have been shown to increase the likelihood of psychiatric disorders, it is clear that medical attention needs to be given to adolescents who use drugs of abuse. It is expected that a decrease in adolescent drug abuse should lead to an accompanying reduction in later psychiatric disorders. PMID- 11028689 TI - Electrophysiologically guided ablation of the pulmonary veins for the curative treatment of atrial fibrillation. AB - Catheter ablation of triggers that induce paroxysms of atrial fibrillation (AF) is an emerging curative therapy for this most common of supraventricular arrhythmias. In a series of 225 consecutive patients with multidrug resistant AF, 96% of triggering foci originated from one or several pulmonary veins (PV) independent of ambient ectopy or structural heart disease. This article describes an ablation procedure that is guided by activation mapping tailored to each individual PV, including criteria to define an arrhythmogenic PV, the use of provocative manoeuvres, the role of circumferential mapping catheters to provide information on the extent, distribution and activation of PV muscle as well as the monitoring of distal PV potentials (PVP) during ablation. Radiofrequency ablation to eliminate distal PVPs is performed by targeting the proximal PVP during sinus rhythm (right PV) or left atrial pacing (left PV). This end-point predicts a successful outcome more often than acute ectopy suppression. Complete elimination of AF is presently achieved in 70% of the patients, resulting in the elimination of antiarrhythmic treatment and suspension of anticoagulant treatment. It is anticipated that continued technological development will improve and facilitate this technique of curative treatment of AF. PMID- 11028690 TI - Increased bronchial nitric oxide production in patients with asthma measured with a novel method of different exhalation flow rates. AB - The concentration of nitric oxide (NO) in exhaled air is increased in patients with asthma, suggesting that measuring fractional exhaled NO concentration (FE(NO)) may be used to monitor asthmatic airway inflammation. However, increased FE(NO) is not specific for asthma, as other inflammatory lung diseases may also increase FE(NO). To augment the specificity of FE(NO) measurement, we tested a novel theoretical modelling of pulmonary NO dynamics that allows the approximation of alveolar NO concentration and bronchial NO flux separately by measuring FE(NO) at several exhalation flow rates. We measured FE(NO) at four exhalation flow rates in 10 steroid-naive asthmatics, 5 patients with extrinsic allergic alveolitis, and in 10 healthy controls. Both the asthmatics and the patients with alveolitis had significantly higher FE(NO) than the healthy controls. The increased NO concentration originated from the bronchial level in the asthmatics and from the alveolar level in the patients with alveolitis. In the second part of the study we assessed the repeatability of FE(NO) test, within day and day-to-day (during two weeks) variation in FE(NO), and the effects of mouth pressure and cigarette smoking on FE(NO) in healthy volunteers. Repeatability of 10 subsequent measurements was high (coefficient of variation (CV) 4.6% +/- 0.4%), and no diurnal variation was found. The day-to-day variation during a 2-week period gave a CV of 10.6% +/- 1.0%. The magnitude of mouth pressure (5-20 cmH2O in adults, 5-40 cmH2O in children) during measurement had no effect on FE(NO). Smoking a cigarette caused a small and transient but statistically significant increase in FE(NO) at 1 and 5 min after smoking. In conclusion, FE(NO) measurement is highly repeatable with low day-to-day variation among healthy subjects. Our results also suggest that the present novel method of measuring FE(NO) at several exhalation flow rates can be used to approximate alveolar and bronchial contributions to FE(NO) separately and thus increase the clinical value of this test. PMID- 11028691 TI - Low serum insulin-like growth factor-binding protein-1 is associated with an unfavourable cardiovascular risk profile in elderly men. AB - Serum insulin-like growth factor-binding protein-1 (IGFBP-1) is regulated by insulin and has been suggested to be a marker of the insulin resistance syndrome. The aim of this study was to investigate whether serum IGFBP-1 is associated with cardiovascular risk factors. Serum insulin-like growth factor-I and IGFBP-3 were also determined. 331 men aged 70-89 years were examined from the Finnish cohort of the Seven Countries Study. Serum IGFBP-1 concentrations were measured by immunofluorometric assay, and the results were analysed with respect to markers of cardiovascular risk, insulin resistance, and insulin secretion that were determined by using the homeostasis model assessment. Fasting serum IGFBP-1 correlated inversely with fasting serum insulin, serum insulin 2 h after oral glucose challenge test, body mass index, and serum triglycerides, and it correlated positively with age and serum high-density lipoprotein (HDL) cholesterol. After adjustment for age, fasting serum IGFBP-1 correlated positively with HDL cholesterol and inversely with fasting serum insulin and triglycerides. Serum IGFBP-1 correlated positively with insulin sensitivity and negatively with beta-cell function. In conclusion, low serum IGFBP-1 levels in elderly men are associated with a number of cardiovascular risk factors. PMID- 11028692 TI - Commending Ernest M. Burgess, MD, PhD with Senate Resolution 278 for his unwavering global service to prosthetics medicine. PMID- 11028693 TI - A device for applying static loads to prosthetic limbs of transtibial amputees during spiral CT examination. AB - This paper describes a device that allows the imaging of prosthetic legs under load conditions using spiral computerized tomography (CT). The device consists of a chair and a vertical footplate mounted on a 17.8 x 1.9 cm oak board that is 197 cm long. The load device can be easily positioned onto the CT bed. A subject sits in the chair and applies force by pushing the foot portion of the prosthesis against the footplate. The magnitude of the force is monitored by a digital force gauge coupled to the footplate. Because the load is borne by the hips and lower back of the subject against the chair, substantial forces can be generated and steadily maintained for the 20-45 s duration of the CT study. This device has been used successfully with 19 transtibial amputees, allowing the acquisition of spiral CT studies with half and full body weight loads. PMID- 11028694 TI - Heterotopic ossification in rehabilitation patients who have had internal fixation of an acetabular fracture. AB - High-energy trauma patients often have multiple injuries and are frequently seen by a physiatrist following their acute care. Acetabular fractures are common in this patient population. Following surgical treatment of acetabular fractures, a very high incidence of heterotopic ossification can occur. We describe 94 patients who underwent posterior surgical fixation of an acetabular fracture. Of these, 87 received heterotopic ossification prophylaxis in the form of irradiation or indomethacin; 5 did not receive prophylaxis. Seven of the 45 patients who were initially started on indomethacin had their medication discontinued for various reasons. Of the 12 patients who did not receive adequate prophylaxis, 5 developed disabling heterotopic ossification. We present our experience with this patient population, and we discuss the importance of adequate prophylaxis for heterotopic ossification. PMID- 11028695 TI - Assessment of spinal movement reduction by thoraco-lumbar-sacral orthoses. AB - Bracing is a common modality in treating spinal fractures. Its objective is to reduce spinal movements and to stabilize the fracture. Until now, factual insight into the movement-reducing properties of Thoraco-Lumbar-Sacral Orthoses (TLSOs) has been missing. Two common TLSOs (e.g., Jewett and Voigt-Bahler) were tested for their movement-reducing properties in two small groups of healthy subjects. In the first study, maximal gross spinal movements, with and without a TLSO, were measured by means of a Portable Posture Registration Set (PPRS) in three different planes. In the second study, maximal segmental vertebral movements in the regions T10 to L4 were measured via X-rays. With few notable exceptions, wearing a TLSO, as measured by the PPRS and X-ray techniques, significantly reduced the segmental as well as gross spinal movements. However, the amount of movement reduction varied greatly from subject-to-subject and was sometimes small. Unfortunately, data are lacking on the amount of movement reduction that is clinically relevant. PMID- 11028696 TI - Experimental and numerical predictions of the ultimate strength of a low-cost composite transtibial prosthesis. AB - Saito et al. (Modern Plastics 1997;74:175-7) have developed a low-cost transtibial prosthesis made of fiber-reinforced plastic (FRP). The prosthesis is comprised of an aluminum pylon, a cosmetic cover, and a constant cross-section composite foot into which aluminum supports are screwed to increase load-bearing capacity. Replacing these supports with a single integrated FRP stiffener significantly reduced manufacturing cost while providing high strength, great durability, and smooth walking. The optimal location and orientation of the proposed FRP stiffener were determined by finite element (FE) analysis. When a replica of this component was tested according to ISO standard 10328, the experimental prosthesis catastrophically failed under 6,600 N of force. Maximum percent error of strain between experimental and numerical results was 18.6%, showing good correlation between the two data sets. The optimized design provides sufficient strength and reduces the cost of manufacturing and thus can be used to replace the original design. PMID- 11028697 TI - Equipment specifications for supported treadmill ambulation training. AB - Supported Treadmill Ambulation Training (STAT) is a mode of therapy for gait retraining for patients with spinal cord injuries or other upper motor neuron dysfunction. The STAT program involves simultaneously supporting a portion of the patient's weight while gait training on a treadmill. STAT has been successful in improving the gait of many research subjects, but has not been widely applied in clinical practice. The goal of this study was to acquire practical, clinically useful information regarding this therapeutic intervention in order to remove barriers to its use. This manuscript enumerates equipment specifications for the treadmill, body weight support (BWS) system, and harness. The ergonomics of the work space are also considered, since the therapist(s) will need access to the patient's legs during therapy. The specific recommendations were determined through prior clinical experience, consultation of anthropometric tables, and application of engineering principles. The guidelines listed are intended to facilitate safe and effective application of the therapy at minimum hardware cost. PMID- 11028698 TI - Electromyographic and kinematic analysis of the shoulder during four activities of daily living in men with C6 tetraplegia. AB - The pattern of motor paralysis that commonly follows C6 tetraplegia creates an increased demand on upper limb function. The present investigation documented shoulder motion and muscular activity during planar motions and four activities of daily living (ADLs) in 15 men with spinal cord injuries (SCI) resulting in C6 tetraplegia. Three-dimensional (3-D) shoulder motion was recorded using a VICON motion system, and intramuscular electrodes recorded electromyographic (EMG) activity of 12 shoulder muscles. Active flexion and abduction required greater EMG than control subjects lifting a 2-kg weight. Relative EMG was similar for most muscles during hair combing, drinking, and reaching forward, although increased humeral elevation commonly resulted in a greater relative muscular effort. Hair combing had the most humeral elevation (90 degrees) with moderate to high levels of activation (32% to 63% maximum) recorded in the anterior deltoid, supraspinatus, infraspinatus, and scapular muscles. During reaching for the perineum, posterior deltoid and subscapularis activity dominated. PMID- 11028699 TI - A compound sensor for biomechanical analyses of buttock soft tissue in vivo. AB - A sensor for in vivo biomechanical characterization of buttock soft tissue has been developed and evaluated. The sensor measures interface pressure, applied force, tilt, and rotation angle of the sensor head, and the thicknesses of multiple soft tissue layers. A composite ultrasonic transducer using a 1-3 ceramic-polymer structure was developed for the sensor. The sensor can identify subcutaneous soft tissue interfaces 5 to 65 mm below the skin surface with a 0.26 mm axial resolution, pressure over the range of 0 to 68 kPa, and force over the range of 0 to 3.6 kg. Our purpose for developing the sensor was to study the biomechanical characteristics of buttock soft tissues. Successful identification of distinguishing characteristics in persons with a susceptibility to pressure ulcers may result in the development of a risk assessment tool based on tissue characterization. PMID- 11028700 TI - Quantifying ataxia: ideal trajectory analysis--a technical note. AB - We describe a quantitative method to assess repeated stair stepping stability. In both the mediolateral (ML) and anterioposterior (AP) directions, the trajectory of the subject's center of mass (COM) was compared to an ideal sinusoid. The two identified sinusoids were unique in each direction but coupled. Two dimensionless numbers-the mediolateral instability index (IML) and AP instability index (IAP) were calculated using the COM trajectory and ideal sinusoids for each subject with larger index values resulting from less stable performance. The COM trajectories of nine nonimpaired controls and six patients diagnosed with unilateral or bilateral vestibular labyrinth hypofunction were analyzed. The average IML and IAP values of labyrinth disorder patients were respectively 127% and 119% greater than those of controls (p<0.014 and 0.006, respectively), indicating that the ideal trajectory analysis distinguishes persons with labyrinth disorder from those without. The COM trajectories also identify movement inefficiencies attributable to vestibulopathy. PMID- 11028701 TI - The effectiveness of neurological rehabilitation in multiple sclerosis. AB - The difficulties inherent in demonstrating the effectiveness of an intervention that is as all-inclusive and poorly defined as neurorehabilitation, especially in a condition as unpredictable and variable as Multiple Sclerosis (MS), are not to be underestimated. They require strict adherence to rigorous methodology and, in particular, the consistent use of a range of clinically appropriate and scientifically sound measures of outcome. Incorporating this approach, it is possible to evaluate rehabilitation at four different levels, including (1) the broadest concept of service delivery; (2) packages of comprehensive care; (3) individual components of the package; and finally, (4) the intrinsic elements of the rehabilitation process. Most recent studies have focused on in-patient rehabilitation and have demonstrated benefits across disability, handicap, and quality of life in patients with mild to severe disability. Such benefits persist following discharge into the community. Studies evaluating service delivery and components of the rehabilitation package are in progress, but few investigators have taken on the intrinsic elements or 'black box' of rehabilitation. These recent studies underline the fact that the evaluation of rehabilitation is feasible. Such studies are important, not simply to justify funding but to ensure continuing improvement of the way in which MS is managed. PMID- 11028702 TI - Microstructural and mechanical characterization of human tissue at and adjacent to pressure ulcers. AB - This investigation evaluated the microstructural and mechanical properties of human skin at and adjacent to pressure ulcers (PUs). Healthy breast and leg tissue served as control tissue. The tissue was characterized through uniaxial tensile testing and histomorphometric analysis. The PU tissue had significantly fewer straight and wavy fibers, but the fibers present were significantly wider and longer than those found in the healthy control tissue. PU ulcer tissue tested in tension had significantly lower strains at peak stress, versus the control breast tissue. Tissue at and adjacent to PUs has undergone significant adaptation or remodeling, as a result of the pressure sustained by the tissue. PMID- 11028703 TI - Fitting hearing aids with the Articulation Index: impact on hearing aid effectiveness. AB - Although most clinical tests focus on how much a particular hearing aid improves speech audibility under controlled conditions, it is unclear how these measures relate to hearing aid effectiveness, or the benefit perceived by the patient under everyday conditions. In this study, the relationship between audibility and hearing aid effectiveness was examined in a cohort of patients who obtained hearing aids through the Veteran's Administration. The measure of audibility was the Articulation Index, a common index of speech audibility. Measures of effectiveness included two hearing-specific surveys and self-reported ratings of global satisfaction and hearing aid use adherence. Results indicated that there were no systematic relationships between measurements of improved audibility and patient ratings of communication ability. Additionally, improved audibility was not related to overall satisfaction with the amplification characteristics of the hearing aid (fitting). However, improved audibility is related to hearing aid use adherence, with patients who achieve better audibility reporting that they use their hearing aids more frequently. PMID- 11028704 TI - The structure and structural effects of VA rehabilitation bedservice care for stroke. AB - The purpose of this study was to: 1) examine the variation in organizational structure within rehabilitation bed-service units (RBU) in the Veterans Health Administration (VHA), and 2) evaluate the effects of RBU and parent hospital structure on stroke rehabilitation outcomes. Two VHA-wide surveys of acute and rehabilitation services for stroke were linked with 2 y of VHA rehabilitation outcomes for stroke patients. A random effects mixed model was used to adjust for patient level covariates, control for unique site effects, and test for facility level structural effects. After adjusting for patient covariates, four structural variables were associated with length of stay or patient functional gain. These results indicate that rehabilitation structure is important to rehabilitation outcome. The individual variables identified in this study, namely, diverse multidisciplinary staff, expert physician leadership, staff participation in team care, and richer rehabilitation equipment resources, may represent the distinct aspects of a successful, comprehensive rehabilitation unit. PMID- 11028705 TI - Is cheap medicine--good medicine? PMID- 11028706 TI - The correction of congenital heart defects with less invasive approaches. AB - BACKGROUND: The minimally and less invasive approaches for the surgical treatment of different heart diseases are rapidly increasing because of their cosmetic and recovery-related advantages. Presented here are the results of less invasive surgery in 51 patients with congenital heart defects. METHODS: From June 1996 to January 1999, we used less invasive techniques for the correction of congenital heart defects in 51 patients. In 32 patients, we performed right anterolateral thoracotomy (6-13 cm), and on the other 19 patients, we used the partial inferior sternotomy (4-7 cm). The ascending aorta and the caval veins were cannulated in all patients. RESULTS: The following congenital heart defects were corrected: ostium secundum atrial septal defect (n = 35), sinus venosus atrial septal defect with partial anomalous pulmonary venous connection (n = 7), ventricular septal defect (n = 7), tetralogy of Fallot (n = 1), and cor triatriatum sinistrum (n = 1). The average age of the patients was 15 years old ranging from 2 months to 48 years and the average weight 39.6 kg (range 3.8-86 kg). The patients were removed from artificial respiratory support on average 8 hours (range 1-48 hours) after surgery and left the hospital after 7 days (range 2-10 days). In 16 patients, blood transfusions were required, an average 5.7 ml/kg BW (range 1.45-19.75 ml/kg BW). The postoperative course was uneventful in all patients. Follow-up (range 3 33 months, mean 17.5 months) was complete with no late deaths or residual defects. CONCLUSION: The right anterolateral thoracotomy and the partial inferior sternotomy provide a safe approach for the correction of certain congenital heart defects. These techniques enable operative correction without any additional risks being incurred and can be performed with standard instruments and cannulation. Additional approaches for extracorporeal circulation are unnecessary. PMID- 11028707 TI - Target, application, and interpretation of scores and alternative methods for risk assessment in cardiac surgery. AB - BACKGROUND: The more popular the use of different methods for risk adjustment becomes, the more often data are applied without any regard about the primary target and/or about important assumptions. Furthermore, risk adjustment is no longer restricted for quality assurance purposes, but became a "tool" of health policy. Few working groups currently use risk adjustment for the development of new therapeutic concepts. The aim of our study is to clarify possibilities and limitations of popular risk adjustment methods. PATIENTS AND METHODS: 4985 Patients underwent isolated CABG. Statistics was performed by calculating descriptive statistics, Parsonnet, and Higginsscores. Furthermore, the parametric, time-adjusted hazard function by Blackstone was used. RESULTS: Descriptive statistics allows intra-, and interinstitutional comparisons of single items to identify "outlying" results. Risk scores aim to predict preoperatively the risk category of the patient who undergoes cardiac surgery. However, since different scores are based on a score-specific combination of variables, and different definitions of the investigation interval, different results may occur, when different scores are calculated for a single patient. However, the use for example, of scores in patient groups allows description of changing risk structures. Most of the scores derive from univariate analyses and monophasic functions. However, survival curves are predominantly multiphasic and require a consideration of the time-dependency of "risk factors". DISCUSSION: An increasing number of patients with severe comorbidity undergoes cardiac surgery. To evaluate reliably present and futurous therapeutic options, risk adjustment is necessary. Since various tools for risk-adjustment are available, a serious discussion about reliability and application is necessary. PMID- 11028708 TI - Evaluation of transmyocardial laser revascularization by following objective parameters of perfusion and ventricular function. AB - BACKGROUND: Does transmyocardial laser revascularization (TMLR), a new surgical technique for treating patients with otherwise intractable angina pectoris, improve myocardial perfusion, metabolism, and, consequently, function? METHODS: Patients referred for TMLR, alone or with coronary artery bypass grafting (CABG), were preoperatively evaluated clinically and by treadmill stress testing, echocardiography, ventriculography, radionuclide assessment of perfusion and metabolism, and hemodynamic assessment. Intraoperatively it was decided that some patients only required CABG. Follow-up evaluations were repeated after 6 (n = 40) and 12 months (n = 23) and compared with preoperative values. RESULTS: CABG only was performed in 35 cases, TMLR + CABG in 17, TMLR only in 45. 1-year mortality was 11% in the TMLR, zero in the TMLR + CABG, and 11% in the CABG groups. In all groups a significantly improved CCS angina- and NYHA class was observed immediately after operation and after 6 and 12 months. In all study groups treadmill tolerance (p<0.05) improved, but regional and global function, perfusion at rest, and metabolism were not significantly changed at 6 and 12 months follow-ups. Perfusion studies under stress demonstrated an improvement only in the CABG group after 12 months (p<0.05), whereas in both TMLR groups the lasered ischemic segments remained unchanged. CONCLUSIONS: TMLR significantly improves long-term clinical status and treadmill stress tolerance, but appears to have little if any effect upon regional and global function, perfusion, and metabolism. PMID- 11028709 TI - Effect of cardiopulmonary bypass and hemofiltration on plasma cytokines and protein leakage in pigs. AB - BACKGROUND: The hypothesis that an inflammatory process during and after cardiopulmonary bypass (CPB) impairs hemodynamics and causes increased capillary protein leakage and that this is possibly ameliorated by hemofiltration (HF) was tested. METHOD: 26 anesthetized pigs were subjected to 120 min CPB (90 min cardioplegia followed by 30 min reperfusion, combined with conventional and modified HF in 13 animals). Hemodynamics, leukocytes, cytokines (IL-1ra, IL-8, IL 10, TNF-alpha), LNPI, plasma protein, and the half-life of i.v. injected Evans Blue (t/2) were assessed before and after CPB. RESULTS: CPB was followed by depression of left ventricular function and activation of inflammatory mediators. Although a slight elimination of some inflammatory mediators occurred, HF did neither improve cardiac function nor reduce the inflammatory process. Plasma protein was lost during CPB and hemofiltration by protein trapping to the surfaces of the CPB system, by filtration across the hemofilter, and by increased microvascular filtration (solvent drag). The latter was probably due to an increased filtration pressure in consequence of the reduction of plasma colloid osmotic pressure by the crystalloid primed CPB. t/2 did not indicate an increased microvascular protein leakage after CPB. CONCLUSION: Hemofiltration is ineffective in improving cardiac function or reducing the inflammatory response of CPB in the pig model. PMID- 11028710 TI - How useful is positron emission tomography for lymphnode staging in non-small cell lung cancer? AB - The introduction of positron emission tomography (PET) raises the question of the new method's capabilities in the staging of mediastinal lymphnodes, since PET differentiates between metabolically active and inactive tissues. 80 patients with histologically confirmed non-small-cell lung cancer (NSCLC) underwent PET scanning with 18-F-marked fluorodeoxyglucose (FDG). Extensive dissection of mediastinal lymphnodes (18-28 lymphnodes recovered) was performed in 78 cases. Metastasis to mediastinal lymphnodes were observed in 25 patients (N2: 22; N3: 3). RESULTS: Primary Tumor: FDG-PET showed significant enhancement of the primary tumor in 78 of 80 patients (sensitivity: 97%). Lymphnode Involvement: FDG-PET was positive in 23 of 25 patients with surgically confirmed lymphnode involvement (sensitivity: 92%). After a median follow up interval of 18 months, 11 patients with false positive lymphnode uptake were still alive; 10 of them showed no tumor recurrency. On the basis of these findings, enlarged mediastinal lymphnodes visualized at CT, but negative at FDG-PET are free of metastatic involvement with a sensitivity of 92%. FDG uptake of mediastinal lymphnodes at PET, however, should not be interpreted as proof of malignancy. PMID- 11028711 TI - FDG positron emission tomography in the diagnosis of peripheral pulmonary focal lesions. AB - Positron emission tomography (PET) using fluoride-18-marked fluoride deoxyglucose (FDG) represents a metabolically based imaging technique capable of providing information on the potential malignancy of peripheral pulmonary focal lesions. In the present prospective study, we investigated the effectiveness of FDG-PET in determining the dignities of 67 such lesions in 35 patients. Findings of FDG-PET were compared with those of computed tomography (CT), as well as with surgical and histological reports, and the value of FDG-PET as a diagnostic method evaluated. FDG-PET correctly identified 38 lesions as positive for malignancy, 18 correctly as negative, 7 incorrectly as negative, and 4 incorrectly as positive. Based on lesions, this yields a sensitivity of 84.4% and a specificity of 81.8%. All malignant focal lesions with a diameter of over 1.2 cm were correctly identified (sensitivity: 100%). In cases of intense FDG uptake, differentiation between a primary lesion, a metastasis, and an acute inflammation is often not possible. PMID- 11028712 TI - Tracheoesophageal fistula after blunt chest trauma: successful diagnosis by computed tomography. AB - A case of tracheoesophageal fistula after blunt trauma is reported. A 27-year-old man who suffered from an automobile traffic accident complained of strong choking after drinking water. Computed tomography demonstrated a defect between the esophagus and the trachea just above the carina. Acquired tracheoesophageal fistula was suspected and promptly confirmed by contrast esophagogram. The defects of the trachea and esophagus was repaired by primary suture and buttressed using a pedicled intercostal flap. The postoperative course was uneventful. PMID- 11028713 TI - Cardiac strangulation in a neonatal case: a rare complication of permanent epicardial pacemaker leads. AB - A newborn female underwent pacemaker implantation with epicardial pacing leads for the treatment of bradycardia caused by congenital atrioventricular block. At 10 months after pacemaker implantation, she was admitted with congestive heart failure. The epicardial leads were seen to encircle the heart, and myocardial ischemia was suggested. During cardiac catheterization she collapsed: emergency operation was performed, but she died on postoperative day 6. PMID- 11028714 TI - Concomitant aortic valve replacement and surgical angioplasty of left main coronary ostium. AB - The association of calcific aortic valve disease and isolated coronary ostial stenosis is rare. A 80-year-old woman was found to have severe aortic stenosis with critical narrowing of the ostium of the left main coronary artery. She was successfully managed by simultaneous aortic valve replacement and patch angioplasty of the left main coronary artery, using a patch of autologous pericardium fixed in glutaraldehyde. Angiographic control at 1 month coupled with intravascular echographic imaging showed adequate relief of the ostial stenosis and patency of the left main trunk. PMID- 11028715 TI - Chronic spontaneous pneumothorax decortication after 130 days. AB - The case of a 21-year-old male with neglected and protracted spontaneous pneumothorax is reported. Video-assisted thoracoscopy after 130 days showed the lung to be trapped solely by a thick pleurovisceral membrane, which required open surgical decortication. Chronic pneumothorax is a rare complication of spontaneous pneumothorax. This is the second report on a chronic pneumothorax solely due to a pleurovisceral membrane and the longest reported interval until decortication. PMID- 11028716 TI - Small thymomas and myasthenia gravis: a poorly understood association. AB - Myasthenia gravis (MG) is associated with about 20% of all patients with thymomas, but the occurrence in patients with small thymomas is rare. The pathogenetic relevance of these small thymomas for MG is poorly understood until now. We report on the clinical course of a 39-year-old female patient suffering from MG and undergoing thymectomy for a small thymoma and discuss the pathogenetic importance of small thymomas for MG. PMID- 11028717 TI - Inflammatory pseudotumor of the lung. AB - Inflammatory pseudotumors of the lung are uncommon in the thoracic surgical literature. These masses resemble carcinoma of the lung radiographically and are discovered only at time the of histologic review of the resected specimen. We report a case of inflammatory pseudotumor that was excised without pulmonary resection, although it completely filled the right hemithorax. We think it was the greatest inflammatory pseudotumor in the English literature. PMID- 11028718 TI - Thymic carcinoma--analysis of nineteen clinicopathological studies. AB - BACKGROUND: Primary thymic carcinomas are rare tumours with nonuniform management protocols and poor prognosis due to delayed diagnosis and highly malignant behaviour. This review summarises the major clinicopathological studies to assess more fully the prognostic importance of tumour histology and staging as well as treatment benefit. METHODS: A Medline search from 1966 to date was carried out and relevant references gleaned from these. Nineteen clinicopathological studies form the core of this review. Demographic data and clinical features were examined in all studies. Correlation of outcome with histology was possible in fifteen of the studies and with tumour staging in thirteen. RESULTS: Thymic carcinomas occur most commonly in the fifth to sixth decades of life, usually presenting with symptoms of space occupation or invasion. Paraneoplastic syndromes are rarely associated, except with the well differentiated thymic carcinoma. Of 140 thymic carcinomas, 40 were squamous cell, 19 were spindle cell and 16 were lymphoepithelioma-like. Completely resected stage I and II tumours demonstrated the best survival. CONCLUSIONS: Complete surgical resection is taken to be the desired treatment, but the continued high mortality and relapse rates and the variable benefit of adjuvant therapies challenge this strategy. PMID- 11028719 TI - Experimental lung transplantation using non-penetrating vascular clips for anastomosis of the pulmonary artery. AB - BACKGROUND: Reliable and rapid anastomosis is critical in lung transplantation. The purpose of this study is to show the appropriateness of sutureless anastomosis of the pulmonary artery using a nonpenetrating vascular clipping device. METHODS: The left lung was excised and the cranial lobe was transplanted autologously in 9 dogs. The anastomosis of the left main pulmonary artery (PA) (1 cm) was performed using a vascular clipping system. RESULTS: PA anastomosis was performed in 7 dogs in 8 to 13 minutes. The endovascular surface of the anastomotic site was smooth, and passage of silicone rubber through the anastomosis was good 14 or 28 days after surgery. Microscopic examination of the anastomotic sites showed excellent healing without evidence of intimal hyperplasia. CONCLUSIONS: The vascular clipping system facilitates anastomosis of the PA in lung transplant surgery. PMID- 11028720 TI - Cholinergic mechanisms of analgesia. PMID- 11028721 TI - The Scandinavian Society of Anaesthesiology and Intensive Care Medicine--from vision to implementation. PMID- 11028722 TI - Adenosine and cardioprotection during ischaemia and reperfusion--an overview. AB - Adenosine is a local hormone, with numerous tissue-specific biological functions. In the myocardium, adenosine is released in small amounts at constant basal rate during normoxia. During ischaemia the production of adenosine increases several fold due to breakdown of adenosine triphosphate (ATP). Increased production of adenosine causes coronary vasodilatation. Thus, adenosine couples myocardial metabolism and flow during ischaemia and is called a homeostatic or "retaliatory metabolite". Furthermore, adenosine has electrophysiological effects in supraventricular tissue, causing a decrease in heart rate. In 1985 it was discovered that adenosine also exerts cardioprotective effects directly on cardiomyocytes. The aim of this review is to give an overview of the role of adenosine as a directly cytoprotective agent during myocardial ischaemia and reperfusion. We will focus on its effects on the myocytes, elicited by stimulation of adenosine receptors in sarcolemma, which triggers intracellular signalling systems. We will also address the new aspect that adenosine can influence regulation of gene expression. There is evidence that the myocardium is capable of endogenous adaptation in response to ischaemia, namely "hibernation" and early and late phases of "preconditioning". Endogenous substances produced during ischaemia probably trigger these responses. We will discuss the role of adenosine in these different settings. Adenosine can be given exogenously through intravasal routes; however, this review will also focus on the effects of endogenously produced adenosine. We will discuss pharmacological ways to increase endogenous levels of adenosine, and the effects of such interventions during ischaemia and reperfusion. Finally, we will review results from studies in humans together with relevant experimental studies, and indicate potential therapeutic implications of adenosine. PMID- 11028723 TI - Efficacy of intrathecal neostigmine for the relief of postinguinal hemiorrhaphy pain. AB - BACKGROUND: Intrathecal administration of various doses of neostigmine has been reported to produce analgesia without neurotoxicity in both animal and human studies. The present study was undertaken to evaluate the efficacy and safety of intrathecal neostigmine for the relief of pain for patients having undergone inguinal herniorrhaphy surgery. METHODS: Sixty men scheduled for elective inguinal herniorrhaphy with spinal anaesthesia were randomly allocated to three groups: group I (n=20) received intrathecal (IT) tetracaine 15 mg, group II (n=20) received IT tetracaine 15 mg+ neostigmine 50 microg, and group III (n=20) received IT tetracaine 15 mg+neostigmine 100 microg. The onset of anaesthesia, duration of analgesia, time to use of first rescue analgesics, the overall 24 h VAS pain scores and the incidence of adverse effects were recorded for 24 h postdrug administration. RESULTS: Onset of anaesthesia (time to T6 sensory block) was significantly faster for group II and III patients compared with group I patients. Motor block (time to lift leg) was greatly prolonged for group III patients, with an average of 6.4 h, compared with 4.1 h for group II patients. Group III patients also showed a later onset of postsurgical pain, lower overall 24-h VAS pain score and prolonged time to first rescue analgesics than did group II patients. There was a significantly greater incidence of adverse effects associated with IT neostigmine, especially nausea and vomiting. CONCLUSION: Our study showed that intrathecal neostigmine at 50 pg or 100 microg enhanced the onset of tetracaine anaesthesia and provided analgesia lasting for 6-9 h, although increased incidences of prolonged motor blockade and nausea or vomiting were noted. PMID- 11028724 TI - Epidural infusion of bupivacaine and fentanyl reduces perioperative myocardial ischaemia in elderly patients with hip fracture--a randomized controlled trial. AB - BACKGROUND: Perioperative myocardial ischaemia is an important risk factor for cardiac morbidity and mortality after noncardiac surgery. The impact of analgesic management on the incidence and severity of cardiac ischemia was studied in 77 elderly patients undergoing surgical treatment of traumatic hip fracture. METHODS: After hospital admission and written consent, patients were randomised to conventional analgesic regimen (intramuscular oxycodone, OPI group) or continuous epidural infusion of bupivacaine/fentanyl (EPI group). The analgesic regimens were started preoperatively. Patients were operated under spinal anaesthesia and the treatments were continued three days postoperatively. ECG was continuously recorded. ST segment depression of > or = 0.1 mV or elevation of > or = 0.2 mV lasting > or = 1 min were considered as ischaemic episodes. Nocturnal arterial oxygen saturation (SaO2) was recorded perioperatively, and subjective pain was assessed every morning using a visual analogue scale (VAS). RESULTS: Fifty-nine (OPI 30, EPI 29) patients were evaluable for efficacy. Thirteen patients (43%) in the OPI and 12 patients (41%) in the EPI group had ischaemic episodes (NS). However, significantly more patients in the OPI group had ischaemic episodes during the surgery (8 vs. 0 in the EPI group, P=0.005). The median (quartal deviation) total ischaemic burden (i.e. integral of ST-change vs. time) in patients with ischaemic episodes was ten times larger in the OPI group (340 [342] mm x min) compared with the EPI group (30 [36] mm x min) (P=0.002). There were no significant differences between the groups in average heart rates or in heart rates at the start of ischaemic episodes or in maximal heart rates during the attacks. Average nocturnal SaO2 was similar in the two groups and there were no differences in the number of hypoxaemic (SaO2<90%) episodes. Preoperatively there were no differences in subjective pain, but postoperative and average perioperative VAS scores for pain were almost 40% lower in the EPI group (P=0.006). Perioperative myocardial infarctions were not detected. CONCLUSIONS: Continuous epidural bupivacaine/fentanyl analgesic regimen, started preoperatively, reduces the amount of myocardial ischaemia in elderly patients with hip fracture. PMID- 11028725 TI - Thoracic epidural analgesia as an adjunct to general anaesthesia for cardiac surgery. Effects on pulmonary mechanics. AB - BACKGROUND: A lasting impairment of pulmonary function is common after cardiac surgery. Pain from the sternotomy may contribute to the impairment. Thoracic epidural analgesia (TEA) can efficiently relieve pain in the postoperative phase, but may also affect respiratory muscle function if local anaesthetics are used. We examined the effects of TEA on pulmonary function and ventilation at rest, before and after coronary artery bypass graft surgery (CABG). METHODS: Thirty patients scheduled for CABG were randomized to receive either general anaesthesia alone or general anaesthesia with TEA. Before and after the operation the patients were examined by respiratory inductive plethysmography and spirometric tests. RESULTS: Before the operation, TEA caused significant reductions in forced vital capacity (FVC), forced expired volume in 1 s (FEV1), maximal inspiratory (PImax) and expiratory (PEmax) pressure. The rib cage contribution to tidal volume decreased significantly but the co-ordination of the thoracic and abdominal movements remained essentially unaffected. Minute volume and respiratory frequency did not change significantly. On the first postoperative day a decrease in maximal breathing efforts was found in both groups. No differences between the groups in FVC, FEV1 and PImax were found, but PEmax was significantly greater in the TEA group. Despite the impairment, breathing at rest was largely normal in both groups. CONCLUSIONS: A better pain-relief from TEA after CABG may improve the ability to cough by a greater expiratory muscle strength. FVC, FEV1, PImax and breathing at rest are not affected by TEA after cardiac surgery. PMID- 11028726 TI - Selective and segmentally restricted antinociception induced by MPV-2426, a novel alpha-2-adrenoceptor agonist, following intrathecal administration in the rat. AB - BACKGROUND: MPV-2426 (radolmidine) is a novel alpha-2-adrenoceptor agonist developed for spinal pain therapy. In the present study we determined the segmental distribution and selectivity of the antinociceptive effect induced by MPV-2426 following i.t. administration in rats. METHODS: The experiments were performed in lightly anesthetized rats with an i.t. catheter for administration of drugs into the lumbar spinal cord level. To determine segmental distribution of antinociception, the withdrawal latency of the tail and forepaw from a hot water bath was measured. To determine selectivity of reflex modulation, the effect of i.t. MPV-2426 on the innocuous H-reflex was determined. RESULTS: In the hot water immersion test MPV-2426 produced a dose-dependent (0.3-3.0 microg) prolongation of tail withdrawal latency whereas the effect on forepaw withdrawal latency was short of significance. Dexmedetomidine, the reference alpha-2 adrenoceptor agonist, produced a significant dose-related prolongation of both the tail and the forepaw withdrawal (0.3 and 1.0 microg). MPV-2426 (1.0 and 3.0 microg) produced no significant change in the amplitude of the H-reflex or M response induced by electrical stimulation of the tibial nerve, nor any change in the modulation of the H-reflex amplitude induced by conditioning sural nerve stimulation. The antinociception induced by MPV-2426 was completely reversed by atipamezole (1 mg/kg s.c.), an alpha-2-adrenoceptor antagonist. CONCLUSION: MPV 2426 produces a selective and segmentally more restricted antinociceptive effect than dexmedetomidine following i.t. administration. The antinoception induced by MPV-2426 is due to action on spinal alpha-2-adrenoceptors. PMID- 11028727 TI - Effects of alpha adrenoreceptor antagonists, prazosin and. yohimbine, on intrathecal lidocaine-induced antinociception in mice. AB - BACKGROUND: The precise mechanisms involved in the spinal analgesic effect of lidocaine are not yet clear. We previously found that lidocaine releases noradrenaline, a modulator of nociception, in rat spinal cord. Here, we attempted to clarify whether or not the noradrenaline release contributes to spinal analgesia by lidocaine. METHODS: The effects of intrathecal injections of the alpha adrenoreceptor antagonists, prazosin (0.01-0.3 nmol) and yohimbine (0.1-3 nmol), on intrathecal 2% lidocaine were assessed using the tail-flick (TF) test in mice. RESULTS: Lidocaine significantly increased the TF latency for 15 min. Prazosin (0.03, 0.1 or 0.3 nmol) and yohimbine (0.3, 1.0 or 3.0 nmol) significantly reduced the lidocaine-induced increase of the TF latency 10 min after injection, although both drugs showed a ceiling effect. CONCLUSION: These results suggest that stimulation of the noradrenergic systems plays an important role in spinal analgesia by lidocaine. PMID- 11028728 TI - Thoracic epidural anaesthesia and analgesia: United Kingdom practice. AB - BACKGROUND: Thoracic epidural analgesia has become increasingly practised in recent years. Complications are rare but potentially serious and, consequently, careful evaluation is required before undertaking this technique. The practice surrounding this procedure varies widely amongst anaesthetists. METHODS: A postal survey to examine the practice of thoracic epidural analgesia was sent to all Royal College of Anaesthetists tutors in the United Kingdom. RESULTS: Responses were received from 240 tutors, representing a return rate of 83%. When obtaining consent for thoracic epidural cannulation, 42% of respondents mentioned risk of a dural tap complication and 11% mentioned neurological damage. Fifty percent of respondents performed epidural cannulation following induction of general anaesthesia. The practice of epidural insertion in patients with abnormal coagulation varied, although over 80% of respondents did not consider concurrent treatment with either aspirin or non-steroidal anti-inflammatory drugs a contraindication. Sterile precautions for epidural insertion also varied between anaesthetists. Postoperatively, 95% of respondents used an opioid-based bupivacaine solution for epidural infusions, and these were most commonly nursed on general surgical wards (63%). Seventy-eight percent of hospitals provided an acute pain team to review epidural analgesia. CONCLUSION: In the United Kingdom, there is little consensus in the practice of thoracic epidural analgesia relating to the issues of informed consent, epidural cannulation in patients with deranged clotting and the sterile precautions taken prior to performing epidural insertion. Most respondents use an opioid-based bupivacaine solution to provide postoperative epidural analgesia. Most hospitals in the UK now provide an acute pain service for thoracic epidural follow-up. PMID- 11028729 TI - Preoperative ropivacaine infiltration in breast surgery. AB - PURPOSE: The aim of the study was to investigate whether preoperative infiltration with ropivacaine in conjunction with breast surgery improves postoperative pain management and attenuates postoperative nausea and vomiting. METHOD: Prospective, randomised, double-blind study, including 60 healthy women (ASA 1-2) allocated to one of two groups. Thirty patients were given 0.3 ml/kg saline in the operating field before surgery. Another 30 patients received a similar volume of ropivacaine 3.75 mg/ml. A visual analogue scale (0-100 mm) was used for evaluation of postoperative pain, nausea and vomiting. If the score was more than 30 mm at rest, the patients were given ketobemidone i.v. as treatment for postoperative pain, and dixyrazine i.v. against nausea and vomiting. The intra-and postoperative analgesic requirements and postoperative nausea and vomiting were registered. RESULTS: The intraoperative fentanyl consumption was similar in the saline group 81 +/- 22 microg vs 76 +/- 28 microg; (ns) in the ropivacaine group. The postoperative 24-h ketobemidone consumption was also similar to those treated with ropivacaine (4.2 +/- 2.6 mg vs 4.2 +/- 4.3 mg; ns). Postoperative nausea and vomiting (PONV) occurred with similar frequencies in both groups. The 24-h dixyrazine consumption was the same in the two groups (2.1 +/- 2.7 mg in the saline group compared to 2.4 +/- 2.8 mg in the ropivacaine group; ns). After 6 h recovery, 41% of all patients had experienced nausea and 20% vomiting. CONCLUSION: We found no differences in postoperative pain management between 3.75 mg/ml ropivacaine and saline wound infiltration before breast surgery. The data show similar postoperative needs of analgesics and antiemetics with a similar frequency of PONV. PMID- 11028730 TI - Pharmacokinetics of 0.2% ropivacaine and 0.2% bupivacaine following caudal blocks in children. AB - BACKGROUND: Ropivacaine is the first S-enantiomer aminoamide local anaesthetic in clinical use, and has been found to be less toxic than bupivacaine. Caudal ropivacine has been shown to cause less motor blockade and longer duration of analgesia in the postoperative period than bupivacaine in children. Plasma levels of ropivacaine and bupivacaine have not been previously compared in children. This study was undertaken to compare the total venous plasma concentrations of similar doses of ropivacaine and bupivacaine following caudal administration. METHODS: Blood samples were obtained to determine the total venous plasma levels of the used local anaesthetic in 30 children, aged 2.3-8.7 years, ASA I, given 1 ml x kg of either 0.2% ropivacaine or 0.2% bupivacaine in a prospective, randomised manner. RESULTS: There were no differences in the individual peak plasma concentrations achieved. Time to the measured peak plasma concentration was significantly shorter in the bupivacaine group. The plasma concentrations of bupivacaine were significantly lower than for ropivacaine at 60, 90 and 120 min after the block. CONCLUSION: Absorption and tissue distribution of ropivacaine is slower than for bupivacaine following caudal administration in children. PMID- 11028731 TI - Mixed venous oxygen saturation during mobilization after cardiac surgery: are reflectance oximetry catheters reliable? AB - BACKGROUND: Oximetry catheters immediately reflect changes in mixed venous oxygen saturation (SvO2). We have used the Baxter 2-SAT system to register changes in SvO2 during early mobilizations after cardiac surgery. To assess catheter reliability, readings were compared to blood gases. METHODS: A total of 352 paired catheter and bench haemoximetry measurements were obtained at the expected highest and lowest levels of SvO2 during the mobilization procedures. The agreement between methods was explored by a Bland-Altman plot. The influence of haemoglobin (Hgb), pH, cardiac output (CO), posture, catheter identity and catheter calibration on agreement was assessed through analysis of covariance. RESULTS: Data included a substantial number of low SvO2 values, 95 paired means of SvO2 < or = 50% and 37 paired means < or = 40%. Mean oxygen saturation difference between catheter and haemoximeter readings was -1.6 +/- 5.7% (SD). Agreement between the methods depended upon the level of SvO2. At SvO2 of 65%, the two methods were virtually identical. Below 65%, the catheters increasingly underestimated the corresponding haemoximetric values by 1.5% for every 10% reduction in SvO2. Agreement was to some degree dependent on individual calibrations and catheter identity, but to a lesser extent on Hgb, CO and posture. CONCLUSION: The two methods are interchangeable for most clinical purposes. Catheter readings are, however, substantially lower than the corresponding haemoximetric measurements at low SvO2 values. Careful interpretation of the absolute values resulting from catheter measurements is recommended, especially when SvO2 readings are low. PMID- 11028732 TI - Effects of oxygen and nitric oxide inhalation in a porcine model of recurrent microembolism. AB - BACKGROUND: Inhalation of nitric oxide (iNO) has been proposed for the treatment of acute pulmonary embolism. The present study evaluates the effects of oxygen (O2) and nitric oxide inhalation in a porcine model of sustained pulmonary hypertension induced by recurrent pulmonary microembolism. METHODS: Twelve pigs were embolized under general anesthesia with 300-microm microspheres intravenously three times over a period of seven weeks. Five pigs served as untreated controls. Hemodynamic and gas exchange responses to 100% oxygen and 40 ppm NO inhalation, and their combination (O2+iNO) were measured seven days after the last embolization. RESULTS: Recurrent microembolism caused sustained pulmonary hypertension (pulmonary vascular resistance index; PVRI 408 +/- 57 dyn x s x cm(-5) x m(-2)) as compared to the control group (PVRI 143 +/- 20 dyn x s x cm(-5) m(-2); P<0.05). PVRI was significantly reduced by O2, iNO, and O2+iNO inhalation by 29 +/- 3, 28 +/- 4, and 32 +/- 3%, respectively. CONCLUSION: We conclude that both O2 and iNO are selective pulmonary vasodilators in a porcine model of sustained pulmonary hypertension following recurrent pulmonary microembolism and, therefore, may be useful in the treatment not only in the acute phase of pulmonary embolism but also later in the time course of the disease. PMID- 11028733 TI - Coagulation effects of a recently developed hydroxyethyl starch (HES 130/0.4) compared to hydroxyethyl starches with higher molecular weight. AB - BACKGROUND: Hydroxyethyl starches (HES) are known to interfere with blood coagulation according to molecular weight, the degree of substitution and the C2/C6 ratio. A recently developed low molecular hydroxyethyl starch (HES 130/0.4) was designed to reduce the blood compromising potency. METHODS: In this study, effects of a 30% in vitro haemodilution with the new HES preparation (HES 130/0.4) in comparison to HES 200/0.5, HES 450/0.7 and sodium chloride solution were investigated using intrinsic and extrinsic activated thrombelastography (TEG) and plasmatic coagulation tests. RESULTS: Whereas plasmatic tests revealed no prolongation of coagulation by HES in comparison to sodium chloride, the TEG variables clotting time, clot formation time and maximal clot firmness showed a significant (P<0.05) inhibition by all the HES preparations. The inhibition was most pronounced in HES 450 (P<0.05 vs HES 130) while HES 130 did not show a statistically significant difference in extrinsic activated maximal clot firmness when compared to sodium chloride. CONCLUSION: These in vitro results demonstrate that hydroxythyl starches especially compromise clot polymerisation. The new preparation HES 130/0.4 seems to inhibit platelet function to a lesser extent than hydroxyethyl starch preparations with a higher molecular weight and degree of substitution. PMID- 11028735 TI - Comparison of the effects of amrinone, milrinone and olprinone in reversing bupivacaine-induced cardiovascular depression. AB - BACKGROUND: Increasing evidence indicates that amrinone, a phosphodiesterase fraction III (PDE-III) inhibitor, has relative efficacy as an initial treatment for cardiovascular depression after bupivacaine in an experimental setting. This study was performed to compare the cardiovascular effects of the other new PDE III inhibitors, milrinone and olprinone, with those of amrinone in reversing bupivacaine-induced cardiovascular depression. METHODS: In sevoflurane anaesthetized dogs, bupivacaine was infused intravenously at 1 mg x kg(-1) x min( 1) until mean arterial pressure fell to 60 mmHg or less. The dogs received either amrinone (2 mg x kg(-1)), milrinone (0.2 mg x kg(-1)), olprinone (0.12 mg x kg( 1)) or 0.9% saline (0.5 ml x kg(-1)). RESULTS: Amrinone, milrinone and olprinone improved left ventricular systolic and diastolic function, resulting in an increase in cardiac index. The most significant difference observed among the three drugs was the change in heart rate (HR) after treatment. Milrinone significantly increased HR, but olprinone did not. CONCLUSION: Milrinone and olprinone are as effective as amrinone in reversing bupivacaine-induced cardiovascular depression. PMID- 11028734 TI - Interaction of isoflurane and cromakalim, a KATP channel opener, on coronary and systemic haemodynamics in chronically instrumented dogs. AB - BACKGROUND: Although isoflurane has been shown to cause coronary and systemic vasodilation through KATP channel activation, the interaction of KATP channel openers and isoflurane has not been fully investigated. The present study was carried out to determine the haemodynamic actions of cromakalim, a KATP channel opener, under the conscious state and during isoflurane anaesthesia in chronically instrumented dogs. METHODS: Fourteen dogs were chronically instrumented to measure systemic and coronary haemodynamics. Each dog was randomly assigned to receive doses of either cromakalim, 4 and 10 microg x kg(-1) i.v., or isoflurane, 2.1% end-tidal (1.5 MAC), plus cromakalim, 4 and 10 microg x kg(-1) i.v. RESULTS: Cromakalim dose-relatedly decreased mean arterial pressure and systemic vascular resistance and increased coronary blood flow in both conscious and anaesthetized states. With isoflurane, the duration of effects of cromakalim were prolonged. Isoflurane exerted an additive effect on the increase in coronary blood flow induced by a low-dose cromakalim, whereas it did not influence the effect of a high-dose cromakalim. The maximum rate of increase in left ventricular pressure and segment shortening were increased by cromakalim in the conscious state but unchanged during isoflurane anaesthesia. CONCLUSION: The results suggest that the coronary vasodilating effects of isoflurane and cromakalim are basically additive until cromakalim exerts the maximal effect, and that the action of cromakalim on the coronary vasculature is prolonged by isoflurane. PMID- 11028736 TI - Influence of opioids on the vascular tone of isolated porcine coronary artery segments. AB - BACKGROUND: It was the aim of this study to elucidate the influence of opioids on coronary vascular tone using the model of isolated porcine coronary artery segments. METHODS: We studied the effects of fentanyl (0.01, 0.1, 1.0 microg ml( 1)), alfentanil (0.1, 1.0, 10 microg ml(-1)), and sufentanil (0.01, 0.1, 1.0 microg ml(-1)) on the contractile response to three vasoconstrictors, acetylcholine, histamine and serotonin. RESULTS: Fentanyl (0.1, 1.0 microg x ml( 1)) dose-dependently attenuated the contractile response to acetylcholine, but not to histamine and serotonin. There were no differences in fentanyl's vasorelaxing potency between rings with intact and denuded endothelium. Alfentanil and sufentanil did not exert any significant influence on any of the vasoconstrictors tested. CONCLUSION: It is concluded that, in isolated porcine coronary artery rings, fentanyl at high concentrations has an attenuating effect on acetylcholine-induced contractions, which is independent of endothelial function, whereas alfentanil and sufentanil do not influence coronary vascular tone. PMID- 11028737 TI - Bispectral index, predicted and measured drug levels of target-controlled infusions of remifentanil and propofol during laparoscopic cholecystectomy and emergence. AB - BACKGROUND: Target-controlled infusions (TCI) have been launched as simple, accurate and reliable delivery systems of intravenous drugs. Bispectral index of EEG (BIS) seems promising in measuring hypnotic effect of anaesthetic drugs. The aims of this study were to evaluate the accuracy of TCI systems in patients undergoing laparoscopic cholecystectomy and to correlate measured drug levels to BIS values. Data were analysed for possible gender differences during emergence. METHODS: After written informed consent, 20 patients were enrolled in an open study. Remifentanil was set at 7.5 ng/ml as target throughout the whole procedure, and propofol at 5 microg/ ml at induction and 3 microg/ml after intubation. Values in blood samples of remifentanil and propofol were correlated to the estimated values and to systolic blood pressure and BIS. BIS values and measured drug levels during emergence and emergence time were compared for the two sexes. RESULTS: Measured drug values varied considerably from the set target with a prediction error of -22% for remifentanil and 49% for propofol. The anaesthesia level was regarded as quite deep with a mean BIS during stable surgery of 42 +/- 7, and at this level we found no correlation between measured values of either of the two drugs and BIS. The emergence time was significantly shorter for women (12.6 +/- 2.5 min) than for men (19.0 +/- 4.2 min) (P=0.001), with no significant differences in measured levels of propofol or remifentanil or BIS during the emergence period. CONCLUSION: Present systems for TCI of remifentanil and propofol result in large intra- and interindividual variations in measured drug levels, and measured levels differ from target. There may be possible interaction between the two anaesthetics at a pharmacokinetic level. Within the level of anaesthesia studied here, BIS was not an indicator of the actual drug levels. Women woke up significantly faster than men. PMID- 11028738 TI - Electrical stimulation of the vestibular system prevents postoperative nausea and vomiting. AB - BACKGROUND: Electrical stimulation of the vestibular system may prevent nausea and vomiting. We studied the influence of transcutaneous impulse stimulation in prevention of postoperative nausea and vomiting (PONV) following gynaecological surgery. METHODS: In this randomised study 70 women undergoing elective gynaecological surgery under general anaesthesia were assigned to receive either the activated (stimulation group) or the inactivated (non-stimulation group) impulse stimulator. The stimulator comprises the stimulator itself, two negative electrodes on a headset applied over both mastoid processes and a nuchal positive electrode. The device yielded a pulse frequency of 5 Hz direct current, individually adjustable between 0.5 and 4 mA. A trapezoid stimulation of 50 ms was applied. Nausea, vomiting, dizziness and the amount of antiemetic drugs used were assessed during the first 4 h postoperatively. RESULTS: Lower postoperative nausea scores with a lower incidence of vomiting and postoperative dizziness were found in the stimulation group. A lower amount of antiemetic drugs was needed in the stimulation group when compared to the non-stimulation group (P<0.01 between groups). CONCLUSION: This study suggests that electrical stimulation of the vestibular system may be useful in prevention of PONV. PMID- 11028739 TI - The effect of desflurane on liver function markers in infants and children. Report of a study and a pertinent case report. AB - BACKGROUND: In an open-labelled clinical trial, the effect of desflurane anaesthesia on liver function markers in paediatric patients was monitored. METHODS: Fifty infants and children, 37 male, scheduled for elective cleft plate surgery were included in the study. Median age was 0.57 (0.25-5.45) years (range), mean desflurane exposure was 2.29 +/- 0.75 MAC-h. Function markers were determined within 24 h prior to and within 24-48 h after anaesthesia. Complete data sets were available for total bilirubin 29, aspartate aminotransferase (ASAT) 36, alanine aminotransferase (ALAT) 35, and for alkaline phosphatase (AP) 28. Pre- and postanaesthetic function tests were compared by means of Wilcoxon's matched-pairs test. RESULTS: Only for AP could a statistically significant reduction of the postanaesthetic values be observed, while the other parameters showed no significant changes. Postanaesthetic ASAT and ALAT were clearly reduced in three children who had unspecific highly elevated preanaesthetic values. After the study, this observation could be repeated in at least one child, who received a further anaesthesia with desflurane within 3 months. CONCLUSION: The data suggest that desflurane does not affect excretory or structural liver integrity in infants and children. PMID- 11028740 TI - Haemodynamic stability during moderate hypotensive anaesthesia for spinal surgery. A comparison between desflurane and isoflurane. AB - BACKGROUND: The aim of this study was to compare desflurane and isoflurane for spinal procedures requiring moderate levels of controlled arterial hypotension, when these agents were administered via a semi-closed circuit at 1 l x min(-1) fresh gas flow. METHODS: After ethics committee approval and written informed consent, 20 ASA I or II patients were randomly allocated to receive either desflurane (n=10) or isoflurane (n=10), in O2/ N2O (1:1) for maintenance of anaesthesia. Induction of anaesthesia, fentanyl dosing and volume loading were standardized. Blood pressure was invasively monitored and maintained within a target systolic blood pressure (SBP) range of 80 to 100 mmHg during the study period. Results were presented as medians and interquartiles, and non-parametric statistical methods were used. RESULTS: Patient demographic data, SBP and heart rate prior to surgery, and duration of the procedure were similar between the two groups. During the study period, tighter arterial blood pressure control was maintained with desflurane as compared with isoflurane. SBP was 21.2% (9.5-41.7) of time outside the range 80-100 mmHg with isoflurane and 5.1% (0.6-10.3) with desflurane (P<0.01). CONCLUSIONS: Desflurane, administered via a semi-closed circuit at 1 l x min(-1) fresh gas flow, maintained better haemodynamic stability in spinal surgery requiring moderate arterial hypotension than isoflurane. PMID- 11028741 TI - Hemidiaphragmatic paresis after interscalene supplementation of insufficient axillary block with 3 mL of 2% mepivacaine. AB - Breathing difficulty, agitation, and confusion developed in a 55-year-old male, ASA classification group III with a non-small-cell lung cancer 10 min after interscalene supplementation of insufficient axillary block with 3 mL of 2% mepivacaine with adrenaline 5 microg mL(-1). After administration of thiopentone and suxamethonium the patient's trachea was intubated and the lungs were ventilated with oxygen-enriched air. The block was successful and surgery was conducted as scheduled. Radiographic monitoring of the lungs at the end of operation showed ipsilateral elevation of the diaphragm with reduced respiratory excursions. Postoperatively, the patient was somnolent and hypercapnic, but maintained satisfactory oxygenation while breathing spontaneously and was extubated. Both the temporal relationship of events and the regression of all symptoms within three hours suggest that 3 mL of mepivacaine with adrenaline injected into the interscalene space blocked the phrenic nerve and compromised diaphragmatic function, which precipitated the respiratory failure. PMID- 11028742 TI - Axillary block studies--quality problems. PMID- 11028743 TI - Evaluation of a commercially available ELISA kit for detection of antibodies to Anaplasma marginale and Anaplasma centrale in cattle in Australia and Zimbabwe. AB - A newly available competitive inhibition ELISA kit for the serological diagnosis of anaplasmosis was evaluated in Australia and Zimbabwe. In Australia the performance of the test was compared with the card agglutination test (CAT). The assay was evaluated using negative sera collected from Anaplasma-free herds, positive sera from experimentally infected cattle and sera from Anaplasma marginale-endemic herds. The sensitivity and specificity of the ELISA in Australia were 100 % and 83,3 %, respectively, and the sensitivity and specificity of the CAT were both 100%. The agreement between the ELISA and CAT in the sera from endemic herds was 86,4 % (kappa = 0,718). The specificity of the ELISA in Zimbabwe was 100%. No meaningful estimate of sensitivity was possible in Zimbabwe because few known positive sera were available for testing, but all eight known positive sera that were available were clearly positive. We conclude that the ELISA is a useful alternative to the CAT for epidemiological studies. The ELISA kits have advantages over the CAT in that the ELISA is more robust and reagents are better standardized, but the kits are expensive. PMID- 11028744 TI - The effect of a natural maedi-visna virus infection on the productivity of South African sheep. AB - A cohort study was conducted in order to measure the effect of the chronic indurative lymphocytic mastitis caused by the South African strain of maedi visna virus (MVV) on the pre-weaning growth of lambs born either of naturally infected or uninfected ewes kept under similar conditions. Fifty naturally infected ewes as well as another 40 from a maedi-visna-free source to be used as control animals, were purchased and kept in separate flocks which were managed in a similar way. All the ewes were of the same breed and 3-4 years old. During the adaptation period, and through the mating, pregnancy and lactation periods they were periodically monitored for the presence of MVV serum antibodies. The lambs were weighed at birth and thereafter every 2 weeks until the age of 90 days, when they were weaned. The ewes were then slaughtered, and their udders examined histologically and the number of lymphocytic follicles were counted and assessed. Although the calculated values indicated a correlation between the number of follicles in the udder and the reduction in the growth rate of the lambs, this was not statistically significant. Similarly, despite higher counts of lymphoid follicles in the udders of sero-positive ewes as compared to those that were sero negative and the lower ewe productivity indexes in infected ewes, no statistically significant differences were found in the indexes of ewes in different follicle categories. PMID- 11028745 TI - Stability and partitioning of closantel and rafoxanide in ruminal fluid of sheep. AB - The stability and the partitioning of closantel and rafoxanide in ruminal fluid (RF) was examined in vitro. Stability was evaluated in two studies in a ruminal fluid-artificial saliva (RF-AS) mixture containing either drug. Drug concentrations were measured in samples collected sequentially from four batches of RF-AS fortified with either closantel or rafoxanide in one study and in four separately incubated aliquots of a RF-AS mixture of each drug in the second study at the start and at various intervals during a 24 h incubation period. The viability of the in vitro RF-AS incubation model was validated by the presence of digoxin degradation (T1/2 of 39,1 +/- 13 h) and by the absence of significant time related differences (P> 0,5) in volume of gas produced, pH and methylene blue reduction time of the RF-AS drug mixture. Partitioning of closantel and rafoxanide was determined by measuring the relative drug concentration of the fluid and particulate phases in RF fortified with either drug at different concentrations. Closantel and rafoxanide were shown to be stable in a RF-AS mixture and were not subjected to any significant biodegradation. An initial marked reduction in drug concentration measured in the RF-AS mixture during the first 2 h of incubation was attributed to the attachment of both drugs onto particulate matter. This was subsequently confirmed in the partitioning study. More than 80% of closantel and rafoxanide was shown to be associated with the particulate phase of RF PMID- 11028746 TI - Morphological changes associated with the development of the rumino-reticulum in growing lambs fed different rations. AB - Morphological changes associated with rumino-reticular development was compared in two groups of SA Mutton Merino lambs (n = 12) fed different diets at 3-5 weeks, 17-19 weeks and 31-33 weeks of age. Diet groups were identified as MMH or MHH according to the sequence at which the full-milk replacement (M) or hay (H) diet were fed to the lambs over the three study phases (phases I, II and III) preceding each age period. Prominent differences in the morphology (size and volume) and function (pH, proteolytic activity and microbial population) of the rumino-reticulum were observed in indicator lambs (n = 2) killed at every age period and also between milk-fed and hay-fed lambs. The size of the rumino reticulum was rudimentary at 3-5 weeks of age and remained underdeveloped in lambs at 17-19 weeks of age which had received a full-milk replacement diet during phase II. One lamb, slaughtered at 3 weeks of age, showed a large distended rumen with severe sloughing of the surface cells of the stratum corneum. The size of the rumino-reticulum increased in size (2 x) in lambs which were fed hay relative to the milk-fed lambs during phase II and reached adult proportions in all lambs at 31-33 weeks of age. Ultrastructural examination showed that rumen papillae were more developed in lambs fed hay during phase II when compared to those of milk-fed lambs. Rumen papillae were best developed in phase III lambs. PMID- 11028747 TI - The rise and fall of tuberculosis in a free-ranging chacma baboon troop in the Kruger National Park. AB - A single troop of free-ranging chacma baboons (Papio ursinus) was found to be infected with tuberculosis caused by Mycobacterium bovis. It is assumed that some members of the troop originally became infected when feeding on a tuberculous carcass in the veld or on tuberculous material scavenged at a nearby post mortem facility. Subsequently, apparent aerosol transmission took place while sleeping in an unused room. Oral transmission probably also occurred due to continuous contamination of the floor of this room and the common, narrow access (a train bridge crossing the Sabi River) to it with faeces and urine. A macroscopic prevalence of 50 % was found and the disease was noted to progress rapidly in infected baboons. A variety of organs had typical tuberculous lesions, of which the spleen, lungs and mesenteric lymph nodes were consistently, grossly affected. Using Restriction Fragment Length Polymorphism analysis, all but one of the baboon isolates were found to be identical to the most common African buffalo (Syncerus caffer) isolate (genotype 1) in this Park. The opportunistic sleeping facility was made inaccessible to the troop, which was forced to revert to sleeping in trees. A follow-up survey six months after closure, demonstrated that the disease had disappeared from the troop, and that no spillover infection had occurred into neighbouring troops. PMID- 11028748 TI - Biochemical and haematological changes in rats administered an aqueous extract of Prunus africana stem-bark at various dosage levels. AB - An aqueous extract of Prunus africana (Hook. f.) Kalkm. (syn. Pygeum africanum) (Hook. f.) (Rosaceae) was administered daily at dosage rates of 10, 100 and 1,000 mg/kg body mass to randomized groups of Sprague Dawley rats. The extract caused a moderate rise in plasma alanine aminotransferase and creatine kinase mainly at rates of 1,000 mg/kg body mass, but it did not cause any significant variations in haematological parameters or in plasma levels of total proteins, albumin, aspartate aminotransferase, alkaline phosphatase and blood urea nitrogen at the dosage levels used. There were no overt clinical signs in any of the rats. It was concluded that the extract may contain components that are mildly toxic to the liver and heart of rats after repeated daily oral administrations of 1,000 mg/kg body mass. PMID- 11028749 TI - Phage types of Salmonella enterica serovar Enteritidis isolated in South Africa from 1991-1995. AB - A total of 615 strains of Salmonella enterica serovar Enteritidis (SE), received from 1991-1995 at the Onderstepoort Veterinary Institute (OVI), were phage typed. Most SE isolates (54,7%) originated from poultry followed by humans (28,5 %) and poultry eggs (9,6 %). Phage type 34 was the most prevalent (40,5 %) of all isolates, followed by phage type 4 (33,8 %). Other phage types identified were 1, lb, 4a, 7, 7a, 9a, 14, 24, 24var and 35 (in total 2,4% of isolates). Most isolates of SE were received from the Western Cape Province (47,4%) and Gauteng (22,3%). In poultry phage type 4 was dominant, but in humans, eggs, goats, ducks, sheep, pigs and rabbits, phage type 34 was the dominant type. It appeared as if the poultry-associated epidemic of SE in South Africa that occurred from 1991 1995 originated in the Western Cape Province during 1991 amongst poultry and then spread from there to humans and eggs and then to the rest of the country, where it emerged during 1993. Results indicate that phage type 34 was the dominant phage type from 1991-1993, but during 1994-1995 its presence declined. During this latter period the presence of phage type 4 increased. This may suggest that two smaller epidemics consisting of the two different phage types might have been responsible for the epidemic that occurred from 1991-1995. PMID- 11028750 TI - Observations on a lesser-known monogenean, Udonella myliobati, from a copepod parasite, Lepeophtheirus natalensis, parasitizing the spotted ragged-tooth shark, Carcharias taurus, from South African waters. AB - The phylogeny of the genus Udonella has been disputed for quite some time, but recent phylogenetic analysis of molecular data confirms that the genus is a monopisthocotylean monogenean. Specimens of Udonella myliobati parasitizing the copepod Lepeophtheirus natalensis, an ectoparasite of the spotted ragged-tooth shark, (Carcharias taurus), were collected and studied. A total of 771 monogenean specimens were recovered from 54 infected copepods examined, with a mean intensity of 14,3 worms per copepod. Most of the monogeneans were found attached to the dorsal surface of the lateral and frontal marginal membranes of the copepod carapace. Eggs, with filaments and adhesive discs, were found ventrally on the host, mainly attached to the maxillae and maxillipeds, in clusters of 12 14. Observations on the morphology, distribution and behaviour of this monogenean are presented with the aid of light and scanning electron microscopy. PMID- 11028751 TI - Selected chemical parameters in the blood and metals in the organs of the Nile crocodile, Crocodylus Niloticus, in the Kruger National Park. AB - Healthy and sick crocodiles of varying sizes were examined from the Olifants River in the central part of the Kruger National Park, the Sabi River in the southern part and the Shingwedzi River in the northern region. Blood was collected for the determination of certain parameters and samples of fat, muscle, kidney and liver tissue were collected and analyzed for their heavy metal content. The results of the blood analyses are within the range recorded in the literature, but the metal analyses were inconclusive as similar data are not available for comparison. The results of the metal analyses are presented here for use as baseline and reference data. PMID- 11028752 TI - Oral infectivity of cryopreserved Haemonchus contortus infective larvae that recovered relatively slowly when thawed after more than 15 years in liquid nitrogen. AB - Infective larvae (L3) of two different batches of anthelmintic-susceptible Haemonchus contortus, cryopreserved for 15,6 and 15,7 years in liquid nitrogen appeared dead when examined 10-20 min after thawing. However, having become motile later in the day, their viability was tested the next day in two separate groups of sheep. The mean development of 41,8% and 66,3% of the H. contortus compared favourably with previous trials with this strain, in which the L3 were injected into the abomasa of sheep after various periods of cryopreservation. It is speculated that the relatively slower recovery rates after thawing of the L3 used in this trial, when compared with those in previous trials in which one of the two batches of susceptible H. contortus was used after shorter periods of cryopreservation, may be an indication that these L3 were gradually approaching the limits of survival after almost 16 years in liquid nitrogen. PMID- 11028753 TI - Occurence of Neodiplozoon polycotyleus Paperna, 1973 (Diplozoidae: Monogenea) in cyprinid fish in South Africa. AB - Neodiplozoon polycotyleus Paperna, 1973 is recorded from Barbus marequensis Smith, 1841, Barbus neefi Greenwood, 1862, and Barbus trimaculatus Peters, 1852 in the Limpopo River System, Northern Province, South Africa. PMID- 11028754 TI - Cyclooxygenase-2: its rich diversity of roles and possible application of its selective inhibitors. AB - In addition to housekeeping cyclooxygenase (COX)-1, which is constitutively expressed in many body cells, an inducible COX-2 has been described and cloned. Induction or presence of COX-2 has been reported not only in isolated cells, but also in cells in various tissues, as well as in both physiological and pathophysiological states, including acute exudative inflammation, proliferative inflammation, animal arthritis, rheumatoid arthritis, angiogenesis, bone absorption, gastric ulcer, colon cancer, hyperalgesia, Alzheimer's disease and certain states of the kidney, brain and female reproductive organs. This review article introduces results from recent works in these fields. COX-1- or COX-2 knockout mice may provide many clues on the roles of COX-2, but may simultaneously cause unnecessary confusion in the recognition of the roles of COX 2, and this is discussed. Recently the roles of COX-2 in exudative inflammation and the anti-inflammatory effects of selective COX-2 inhibitors have been questioned. This is discussed in the text. Prostanoids mediate signals to adjacent cells to provide fine regulation of cellular function. Because of the short duration of the expression of COX-2 gene and protein, COX-2 must play some roles different from those of COX-1 gene and protein in vivo. It is not yet possible to identify all the roles of COX-2, but in some tissues, such as the kidney, the brain and others, COX-2 may be expressed constitutively, whereas the prostaglandin generation by COX-2 may replace that by COX-1 in some states (or vice-versa). Precise analyses of the expression of COX-2 may disclose fine modulation of cellular and organ functions by PGs. Several selective or preferential COX-2 inhibitors have been developed and were shown to be effective in clinical trials. Most were reported to be free of adverse gastrointestinal effects, but it should be noted that in the healing process of gastric ulcers and in sodium-restricted states, adverse effects of selective COX-2 inhibitors could be expected. Soon, with more detailed knowledge of the delicate roles of COX-2 in vivo, effective and safe application of COX-2 inhibitors should be realized. PMID- 11028755 TI - Both interferon (IFN)alpha and IFNgamma inhibit histidine decarboxylase expression in the HT168 human melanoma cell line. AB - OBJECTIVE: Histamine plays an important role in a series of processes including inflammation, allergy, gastric acid secretion, neurotransmission, embryogenesis and in various tumours. Histidine decarboxylase (HDC), the enzyme solely responsible for generation of histamine is expressed in many cells including regenerating and tumour cells. HDC expression is regulated by multiple tissue factors, e.g. various cytokines and growth hormones. In this study the effect of interferon alpha and interferon gamma on the expression of HDC and on cell proliferation in vitro on melanoma cell line. METHODS: We used recombinant human interferon alpha, interferon gamma and human melanoma cell line HT168. RESULTS: Our data show that both IFNalpha and IFNgamma decreased the HDC mRNA and protein expression, though with dissimilar kinetics. IFNgamma strongly suppresses the proliferation at 72 h, while IFNalpha has a more moderate effect. CONCLUSIONS: Since previously the inhibitory effect of histamine on gene expression of interferon gamma was detected, a reciprocal inhibition between histamine and IFNgamma is proposed. PMID- 11028756 TI - l-Ephedrine is a major constituent of Mao-Bushi-Saishin-To, one of the formulas of Chinese medicine, which shows immediate inhibition after oral administration of passive cutaneous anaphylaxis in rats. AB - OBJECTIVE AND DESIGN: Whether Mao-Bushi-Saishin-To (MBST), one of the formulas of classical Chinese medicine, is effective on 48-h passive cutaneous anaphylaxis (PCA) in rats and which substance in the formula is responsible for its inhibitory action were examined. TREATMENT: In the studies on PCA, MBST (hot water extract of the whole herbal formula), extracts of Ephedra herb (Mao), l ephedrine and other reference drugs were orally administered immediately or at various times before or 5 min after the antigen challenge. In the experiments on anaphylactic histamine release from rat peritoneal mast cells, l-ephedrine and d pseudoephedrine were added at 10(-4)-10(-7) g/ml at 30, 10, 3 or 0 min before antigen provocation. RESULTS: The time course study indicated that MBST produced a prompt and long lasting inhibition of PCA. Among the constituents of Mao, l ephedrine exerted this prompt inhibitory activity, but d-pseudoephedrine did not. Neither pseudoephedrine nor l-ephedrine prevented the anaphylactic histamine release from isolated peritoneal mast cells. CONCLUSIONS: It is strongly emphasised that the rapid suppression of PCA by orally administered l-ephedrine must be exerted by a mechanism distinct from that of suppression produced following gastrointestinal absorption of the drug, because the time required for the inhibition was extraordinarily short. However, direct inhibition of anaphylactic histamine release from isolated mast cells was excluded in this inhibition of PCA. PMID- 11028757 TI - Effect of FTY720, a novel immunosuppressant, on adjuvant-induced arthritis in rats. AB - OBJECTIVE AND DESIGN: Anti-arthritic effect of FTY720, a novel immunosuppressant, was compared with those of immunosuppressants cyclosporin A and tacrolimus in adjuvant-induced arthritis in rats. MATERIAL: Male LEW rats. TREATMENT: FTY720 (0.03-0.3 mg/kg), cyclosporin A (1-10 mg/kg) or tacrolimus (0.3-3 mg/kg) were orally administered to rats for 21 days beginning on the day (day 0) of adjuvant inoculation. In addition, the anti-arthritic effect of FTY720 (0.3 mg/kg) and cyclosporin A (10 mg/kg) were evaluated by administration to animals for 5 consecutive days (days 2-6, 6-10, and 10-14). METHODS: Adjuvant-induced arthritis was produced by intradermal injection of 0.5 mg heat-killed Mycobacterium tuberculosis. Hindpaw edema was measured plethysmographically. The day of arthritis onset was determined macroscopically. Bone degradation was determined by radiography. Peripheral blood leukocytes were classified microscopically. RESULTS: All test compounds inhibited the incidence of arthritis, hindpaw edema and bone destruction. In addition, FTY720 but not cyclosporin A or tacrolimus markedly decreased the number of peripheral blood lymphocytes. FTY720 treatment on days 6 to 10 inhibited the bone destruction and hindpaw edema. CONCLUSION: These results suggest that the anti-arthritic effect of FTY720 in this adjuvant induced arthritic model was more potent than those of cyclosporin A and tacrolimus. FTY720 administered on days 6 to 10 showed the inhibitory effect on the bone destruction and hindpaw edema. FTY720 may be effective in the treatment of rheumatoid arthritis. PMID- 11028758 TI - Nitric oxide inhibits leucocyte migration in carrageenin-induced rat pleurisy. AB - OBJECTIVE AND DESIGN: The role of nitric oxide (NO) on leucocyte migration has been investigated in rat carrageenin-induced pleurisy. MATERIAL: Male Wistar rats. TREATMENT: L-arginine, NOC-18 and aminoguanidine were administered subcutaneously 1 h prior to carrageenin injection. METHODS: Leucocyte accumulation into the pleural cavity was measured 4 h after carrageenin challenge. Statistical significance was calculated by Bonferroni test. RESULTS: L arginine (10 mg/kg) or the NO donor NOC-18 (10 mg/kg), significantly inhibited leucocyte infiltration by 31% and 20% respectively (P<0.01). On the contrary, when these compounds were given at high doses (L-arginine 300 mg/kg; NOC-18 30 mg/kg), leucocyte accumulation was increased by 22% and 33% respectively (P<0.01). Aminoguanidine, a relatively selective inhibitor of the inducible NO synthase, depending on the dose, showed a biphasic effect on cell migration. Thus, at low doses (30 and 100 mg/kg), aminoguanidine increased (by 40% and 74% respectively, P< 0.01) leucocyte infiltration which was inhibited by 41% (P < 0.01) when the drug was given at high dose (300 mg/kg). CONCLUSIONS: These results suggest that in rat carrageenin-induced pleurisy NO primarily inhibits leucocyte migration. PMID- 11028759 TI - An experimental study in the chick embryo chorioallantoic membrane of the anti angiogenic activity of cyclosporine in rheumatoid arthritis versus osteoarthritis. AB - OBJECTIVE AND DESIGN: Angiogenesis plays an important role in the pathogenesis of rheumatoid arthritis (RA) and correlates with clinical score, synovial hyperplasia and infiltration of inflammatory cells. Many of the available treatments for RA have been shown to possess some degree of anti-angiogenic activity. Here, we studied the effect of cyclosporine, which exerts anti angiogenic activity in vitro and in vivo [1] on angiogenesis induced in vivo in the chick embryo chorioallantoic membrane (CAM) by synovial RA and osteoarthritis (OA) tissues. MATERIAL AND METHODS: Wet synovial biopsies from 10 RA and 6 OA patients were treated with vehicle alone or with cyclosporine and implanted on the CAM at day 8 of incubation. On day 12, CAM tissues were assessed for the extent of angiogenesis and mononuclear cell infiltration. RESULTS: Cyclosporine inhibited angiogenesis and reduced the number of mononuclear cells in the CAM extracellular matrix only in RA implants. CONCLUSIONS: These data provide further evidence for a central role of new-formed blood vessels in RA. Moreover, cyclosporine on account of both its immunosuppressive and its anti-angiogenic activity can be proposed for the treatment of RA. PMID- 11028760 TI - High-dose clodronate therapy prevents joint destruction in chronic antigen induced arthritis of the rat but inhibits bone formation at the axial skeleton. AB - OBJECTIVE: To investigate the effects of clodronate on clinical disease activity, inflammatory alterations and cartilage destruction, periarticular and axial bone volume and bone turnover in chronic antigen-induced arthritis (AIA; day 28). METHODS: Rats with AIA were treated with clodronate (5 mg/kg/day continuously; 20 mg/kg/day intermittently or high-dose with 300 mg/kg 3 hours after arthritis induction +20 mg/kg/day continuously, respectively). Joint pathology was examined by histology. Bone volume and cellular turnover parameters of the right tibia head and the third lumbar vertebra were evaluated by histomorphometry. The findings were compared with those of healthy controls, sham-treated AIA and AIA treated continuously with 250 microg/kg of dexamethasone. RESULTS: All three therapy regimens with clodronate resulted in a significant reduction of joint swelling, histopathological inflammatory changes and cartilage destruction in comparison with sham-treated AIA. The antiinflammatory effect of high-dose clodronate was comparable with dexamethasone. The intermittent administration of 20 mg/kg/day of clodronate completely prevented periarticular bone loss by reduction of bone resorption without affecting bone formation at the periarticular and axial bone. Both continuous treatment with 5 mg/kg/day of clodronate and high-dose clodronate therapy partially prevented periarticular bone loss and reduced parameters of bone formation at the axial bone to values below those of healthy controls. CONCLUSION: High-dose clodronate therapy exerts an excellent preventive effect on clinical disease activity and on joint destruction in AIA. However, continuous treatment with high doses of clodronate may result in a low turnover state of bone remodelling. PMID- 11028761 TI - The science of systematic reviewing studies of diagnostic tests. AB - BACKGROUND: Systematic reviews have gradually replaced single studies as the highest level of documented effectiveness of health care interventions. Systematic reviewing is a new scientific method, concerned with the development and application of methods for identifying relevant literature, analysing the material while increasing validity and precision, and presenting and discussing the results in a way that does justice to the research question and to the available evidence. The objective of this study was to review the systematic reviews in laboratory medicine, to evaluate the methods applied in these reviews and the applicability of guidelines of the Cochrane Methods Working Group on Screening and Diagnostic Tests, and identify areas for future research. METHODS: All the systematic reviews in the field of clinical chemistry and laboratory haematology that could be identified in Medline, EMBASE and other literature databases up to December 1998, were evaluated. RESULTS: We studied 23 reviews of diagnostic trials. Although all reviews share the same basic methodology, there was a wide variation in the methods applied. There was no consensus on the quality criteria for inclusion of primary studies. The results of the primary studies were heterogeneous in most cases. This was partly due to design flaws in the primary studies, but was also inherent in the diverse study designs in diagnostic trials. We observed differences in the analysis of the factors that cause heterogeneity of the results, and in the summary statistics used to pool the data from the primary studies. The additional diagnostic value of a test, after other test results are taken into consideration, was only addressed in one study. CONCLUSION: This overview of 23 reviews of diagnostic trials identifies areas in the methods of systematic reviewing where consensus is lacking, such as quality rating of primary studies, analysis of heterogeneity between primary studies and pooling of data. Guidelines need to be improved on these points. PMID- 11028762 TI - Need for revisiting the concept of reference values. AB - The reference values concept has been adopted by health care professionals, including clinical chemists, laboratory scientists, and clinicians and simultaneously by all the official organizations in charge of the establishment of legislation. But the estimation of reference limits, and the evaluation of biological variability need to be improved at the level of the procedures, which are currently too long and too expensive and not feasible easily for all laboratories. The procedures for obtaining reference values, if we follow the original documents, are complex, and that is the main reason that clinical chemists or diagnostic kit manufacturers have not used them systematically. There is clearly a need that scientific societies and international organizations propose practical recommendations: 1) Recommendations to describe methods linked to systematic error. * How to transfer reference limits from one laboratory to another laboratory using different methods? * Should we determine reference limits for each method? * How can we differentiate bias due to the populations from these due to the method? Clear collaborations with manufacturers involved in kits and diagnostic systems are needed. 2) Practical recommendations linked to the reference population. * How to select a homogeneous population? (Careful recommendations on the choice between healthy individuals, blood donors and individuals hospitalised for other diseases should be given.) * How to estimate ethnic differences? * How to define the exclusion and inclusion criteria according to quantity? * How to deal with the question of reference limits for unstable periods, aging or old people particularly, when the difference between aging and disease is very difficult to define? 3) Practical recommendations on the statistical methods to be used. * How to make a good choice of the interquartile interval? Should we use and present only the centiles 2.5 or 97.5, or on the contrary should we give other centiles in addition, for example 5, 10, 75, 80, 85, 90? 4) Practical recommendations linked to the use of the concept of the reference values. * How to make this concept more concrete and to have official definitions which are better understandable and not only abstract? * How to demonstrate the value of using simultaneously reference limits and decision limits, and what does each of these limits bring to results interpretation? * How to improve the presentation of the results? How to give more information on biological variability in the laboratory data, taking into account the scientific validity of their determination? Should we use new information techniques and new communication systems for reaching these objectives? The responses to all these questions could only be provided if there is a concerted effort at the international level. Practical recommendations should be given, which would be very useful for a better understanding and use of reference values by laboratory scientists and clinicians. PMID- 11028763 TI - Purification of anti-thyroglobulin IgG from human serum. AB - The high prevalence of subjects in the general population and among patients with thyroid diseases who are positive for serum anti-thyroglobulin antibodies, together with the inconsistency of hybridoma techniques to obtain large amounts of anti-thyroglobulin antibodies, prompted us to prepare anti-thyroglobulin antibodies from human serum. An anti-thyroglobulin antibody positive serum was absorbed with CNBr-activated sepharose 4B conjugated to thyroglobulin and anti thyroglobulin antibodies eluted by acid pH (3.0), basic pH (10.7) and detergent (SDS 3 g/l or 7 g/l). Preliminary experiments carried out with rabbit anti-human thyroglobulin allowed us to purify an Ig-fraction that retained its anti thyroglobulin activity by immunoprecipitation and tanned red cell haemagglutination. When subjected to agarose gel electrophoresis and Silver stain, this fraction has the mobility of rabbit lg. Further experiments were carried out using Hashimoto's serum (tanned red cell haemagglutination 1:40620). pH 3.0 was found to give the best yield of stable antibodies. An IgG fraction was eluted at a concentration of 10+/-1.2 mg/l of serum. This fraction has the same electrophoretic mobility as human IgG and is close to pure human anti thyroglobulin antibody. A dose-response curve was built up at a concentration range between 0.016 mg-2 mg of IgG anti-thyroglobulin antibody per litre. The slope of the curve parallels that of a dilution curve of the whole serum, suggesting that the purified antibodies are representative of the whole antibody population. In conclusion, we provide a method for preparing considerable amounts of highly purified anti-thyroglobulin antibodies from human serum for application in clinical medicine and basic research, and in particular which provides a standard for measuring anti-human thyroglobulin serum antibodies by weight rather than the presently used standard expressed in U/ml. Furthermore, this reagent could act as an ideal immuno-vector for the diagnosis and therapy of differentiated thyroid carcinoma. PMID- 11028765 TI - Sequential C3 and C4 levels in human milk in relation to prematurity and parity. AB - Similarly to many immune molecules of human milk, C3 and C4 levels decrease during lactation. We investigated the influence, over the first three weeks of lactation, of both prematurity and parity on the sequential evolution of these levels. Milk C3 and C4 concentrations were measured by immunonephelometry in 494 individual samples collected from 76 lactating mothers. C3 and C4 concentrations were higher in milk from preterm or primiparous mothers. The major differences were observed in milk from days 5-8 and 9-20, likely due to pronounced interindividual variations in levels of days 1-4 milk. Milk from mothers of precocious (33 weeks' gestation or less) preterm newborns presented higher concentrations and a slower decrease of C3 and C4 levels than that from mothers of late (33-37 weeks' gestation) preterm newborns, when compared to term mothers. Finally, the inversion of the C3/C4 ratio occurring over time, previously reported, appeared later in milk from mothers of preterm newborns. The influence of prematurity was even greater in primiparous than in multiparous mothers. Both C3 and C4 levels therefore appear to be influenced in human milk by the parity and prematurity of the delivery. Mothers from preterm newborns seem to provide higher levels of C3 for a longer period post delivery. PMID- 11028764 TI - Preeclampsia is associated with alterations in insulin-like growth factor (IGF)-1 and IGF-binding proteins in Wharton's jelly of the umbilical cord. AB - Wharton's jelly is abundant in extracellular matrix, which is known as a storage site to concentrate and stabilise growth factors in the vicinity of cells. It was previously found that Wharton's jelly contains significant amounts of insulin like growth factor (IGF)-1 and IGF-binding proteins (BPs). IGF-1 is a stimulator of biosynthetics of collagen and sulphated glycosaminoglycans. Preeclampsia (edema, proteinuria, hypertension (EPH)-gestosis) is accompanied by an accumulation of sulphated glycosaminoglycans in Wharton's jelly. IGF-1 and BPs may play an important role in such a remodelling of this tissue. It was decided to evaluate the alterations in amounts of IGF-1 and BPs in Wharton's jelly of newborns delivered by mothers with preeclampsia. Studies were performed on Wharton's jelly of 10 controls and 10 newborns delivered by mothers with preeclampsia (edema, proteinuria > 500 mg/l, arterial pressure: systolic > 140 mm Hg, diastolic > 90 mmHg). Radioimmunological techniques were employed to determine IGF-1 and IGF-BPs (BP-1 and BP-3). It was found that preeclampsia is associated with a decrease in IGF-1 and IGF-BP-1 in Wharton's jelly. A slight increase in IGF-BP-3 was found. Ligand blotting demonstrated that BP-3 (not BP-1) is a main component of Wharton's jelly, which binds IGF-1. Heparin drastically inhibited the binding of IGF-1 by BP-3. It is known from our previous studies that preeclampsia is associated with an increase in the amount of sulphated glycosaminoglycans (heparin, heparan sulphate, dermatan sulphate) in Wharton's jelly. This may be a factor, which prevents the binding of IGF-1 by BPs and facilitates the binding of IGF-1 to cells, stimulating them to produce sulphated glycosaminoglycans in Wharton's jelly. PMID- 11028766 TI - Evaluation of Emit tox benzodiazepine and barbiturate assays on the Vitalab Viva analyser and FPIA on the Abbott ADx analyser. AB - We evaluated the performance of the Emit tox benzodiazepine and barbiturate assays (Dade Behring) and fluorescence polarisation immunoassay (FPIA) (Abbott) for use with serum determinations in preliminary therapeutic drug monitoring or acute drug intoxication detection. Performance, as indicated by CVs, of the Emit tox benzodiazepine and barbiturate assays and FPIA showed that both immunochemical techniques are precise and have good reproducibility. For within run studies, results from benzodiazepine determinations showed maximum CV values of 1.91% for the Emit method and 2.65% for FPIA; results from barbiturate determinations showed maximum CV values of 2.01% for the Emit method and 1.89% for FPIA. For between-run studies, results from benzodiazepine determinations showed maximum CV values of 1.79% for the Emit method and 1.12% for FPIA; results from barbiturate determinations showed maximum CV values of 2.09% for the Emit method and 2.02% for FPIA. PMID- 11028767 TI - Serum eosinophil cationic protein in active and quiescent ulcerative colitis. AB - Inflammatory bowel disorders are characterized by an accumulation of eosinophilic granulocytes, mast cells, lymphocytes and neutrophilic granulocytes in the intestinal mucosa. The aim of this study was to examine the concentration of eosinophilic granulocytes in the blood of patients during active ulcerative colitis in comparison with patients during remission and apparently healthy control subjects. Besides counting, the activity grade of eosinophilic granulocytes has been studied by estimation of their degranulation product eosinophil cationic protein. Subjects with active ulcerative colitis could be distinguished from patients with quiescent ulcerative colitis by establishment of the eosinophil cationic protein concentration, neutrophilic granulocyte count, erythrocyte sedimentation rate, C-reactive protein and albumin concentration. After two weeks of corticosteroid treatment, serum eosinophil cationic protein concentrations and eosinophil counts in blood were significantly decreased. A decrease in blood eosinophil count was accompanied by a decrease in eosinophil cationic protein concentrations in serum in most subjects with ulcerative colitis. After twelve weeks of corticosteroid administration, serum albumin concentrations were significantly increased, whereas serum concentrations of C reactive protein were significantly decreased. During treatment with corticosteroids, serum eosinophil cationic protein concentrations and blood eosinophil counts are appropriate laboratory markers to detect the effect of medication in the course of ulcerative colitis. PMID- 11028768 TI - Measurement of purines in urine by capillary electrophoresis for estimating the degree of hypoxia in infants. AB - We describe a simple, rapid and reproducible method for measuring hypoxanthine, xanthine and uric acid in urine samples of infants by capillary electrophoresis with an uncoated fused silica capillary and ultraviolet detection. Conditions were 40 mmol/l borate buffer pH 9.4 with sodium dodecyl sulphate (0.1 mol) 3 s hydrodynamic load and the voltage 20 kV. The calibration curves for hypoxanthine and xanthine were linear to 150 micromol/l and for uric acid to 300 micromol/l. The limits of detection for hypoxanthine, xanthine and uric acid were 0.5, 2.0 and 20 micromol/l, respectively. Analytical recovery of the three purines ranged from 93 to 105%. Overall CVs were < 5.2%. Using this method, we measured purine concentrations in the urine samples of 103 infants, divided into three groups, according to different degrees of hypoxia. There was a significant difference among the three groups only in hypoxanthine concentrations. In 35 urine samples from the control group, 43 urine samples from infants with a history of an apparent life threatening episode and in 25 urine samples from infants with infantile apnoea, the mean concentrations of hypoxanthine (mean +/- SD) were 13.1 +/- 4.0, 25.1 +/- 8.1 and 58.2 +/- 17.4 micromol/mmol creatinine, respectively. The diagnostic accuracy of the purine measurement was evaluated by receiver operating characteristic curve analysis. PMID- 11028769 TI - Separation of erythrocytes into age-related fractions by density or size? Counterflow centrifugation. AB - During the process of aging red blood cells become denser and smaller. Counterflow centrifugation separates particles of lower density and smaller diameter from those of higher density and bigger diameter. Thus, the question arises: which property of the red cells, density or size, governs the age-related separation by counterflow centrifugation? It is shown that it is the size which dominates the balance between sedimentation and streaming. Age-related separation of human red blood cells by counterflow centrifugation (elutriation) was analysed by the standard hematological parameters (hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration), hemoglobin A1c and the membrane protein ratio 4.1a/(4.1a+4.1b). Red blood cells with a high hemoglobin A1c content and a high 4.1a/(4.1a+4.1b) ratio were found in the early fractions of the elutriation. This proves that old cells make up early fractions, while the "youngsters" constitute later ones. The elutriation technique used (yielding human red blood cells in a "healthy state") and the age parameters studied show that the membrane protein ratio 4.1a/(4.1a+4.1b) is another reliable age parameter for the assessment of red blood cell age. PMID- 11028770 TI - Can age and sex related reference intervals be derived for non-healthy and non diseased individuals from results of measurements in primary health care? AB - Reference intervals in clinical chemistry are commonly based on results of measurements in reference populations or are taken from the literature. A reference population should represent a defined group of individuals and be as similar as possible to the patients under investigation. Frequently, reference populations have been recruited from institutionalised healthy young people who do not necessarily fulfill these criteria. In the present study we describe the temporal changes in 37 commonly measured quantities in men and women from childhood to late in life. The samples were collected in the primary health care and sorted according to an assumed decision by the physician. The emerging group of individuals forms a reference population that was regarded as "non-diseased" and the results of measurements in this population are reference values. A remaining group of "non-healthy" were likewise identified for comparison. The central 95 percentile was wider than those usually assigned to the quantities whereas the medians almost coincided. In the "non-healthy" group the medians were shifted in a direction that would be expected from pathophysiology aspects. PMID- 11028771 TI - Associations of apolipoprotein E concentration and polymorphism with lipids and apolipoprotein levels in Chinese from Beijing and Shanghai. AB - The association of apolipoprotein E concentration and common polymorphism (codons 112/158) with lipid and apolipoprotein concentrations was studied in two Chinese healthy population samples from Beijing (n=99) and Shanghai (n=67). Body mass index and apolipoproteins E and AI and triglyceride concentrations were significantly different between the two populations (0.0010.05). The specificity and sensitivity of pleural uric acid for diagnosis of transudate effusions were 73% and 80.6%, respectively. The specificity and sensitivity of pleural uric acid for diagnosis of transudate effusions from exudates without malignancy were 71.8% and 91.7%, respectively. The sensitivity and specificity of pleural lactate dehydrogenase for diagnosis of exudates were 82% and 89%; the sensitivity and specificity of pleural fluid/serum lactate dehydrogenase were 85% and 89%; the sensitivity and specificity of pleural fluid/serum protein were 91% and 89%, respectively. Using all three of Light's criteria together, the sensitivity was 91% and its specificity was 94%. Our findings indicate that determination of uric acid in pleural fluid may be of diagnostic value in differential diagnosis of transudates and exudates. The sensitivity of pleural uric acid measurement was higher for exudates without malignancy. However, Light's criteria remain the best means of separating transudates from exudates. PMID- 11028773 TI - Evaluation of the Roche Diagnostics LightCycler-Apo B 3500 Mutation Detection Kit. AB - Familial defective apolipoprotein (apo) B-100 is an autosomal codominant disorder associated with hypercholesterolemia and an increased risk of coronary artery disease. Two independent mutations affecting the codon 3500 (Arg3500-->Gln and Arg3500-->Trp) have been shown to cause ligand-defective apo B-100. Identification of carriers of these mutations is an important step in the risk stratification of individuals and families with hypercholesterolemia. We evaluated a homogeneous assay for detection of mutations at codon 3500 that combines rapid-cycle PCR with allele-specific fluorescent probe melting profiles for product genotyping. This single-tube analysis is performed on the LightCycler, a microvolume fluorimeter integrated with a thermal cycler. Continuous acquisition of fluorescence data during a melting curve analysis at completion of PCR allows the detection of mutations, as loss of fluorescence occurs in an allele-specific manner. By plotting melting peaks, the three apo B 100 alleles were readily distinguishable. Using this method, genotyping of 32 samples is completed within 40 min without the need for any post-PCR sample manipulation, thereby eliminating the risks of end-product contamination and sample tracking errors. The specific detection of mutations at codon 3500 of the apo B gene on the LightCycler is a rapid and reliable method that is ideally suitable for typing both small and large numbers of samples. PMID- 11028774 TI - HIV infection does not explain elevation of glycated hemoglobin among non diabetic patients in Gabon. PMID- 11028775 TI - Plasma and serum total homocysteine concentrations in paediatric patients, evaluated by high-performance liquid chromatography with fluorescence. PMID- 11028776 TI - Systematic synthesis of sulfated sialyl-alpha-(2 --> 3)-neolactotetraose derivatives and their acceptor specificity for an alpha-(1 --> 3) fucosyltransferase (Fuc-TVII) involved in the biosynthesis of L-selectin ligand. AB - Sulfated sialyl-alpha-(2 --> 3)-neolactotetraose (IV3NeuAcnLcOse4) derivatives at C-6 of GlcNAc (6-O-sulfo), terminal Gal (6'-O-sulfo), and both GlcNAc and Gal (6,6'-di-O-sulfo) residues have systematically been synthesized. (Methyl 5 acetamido-4,7,8,9- tetra-O-acetyl-3,5-dideoxy-D-glycero-alpha-D-galacto-2 nonulopyranosy lonate)-(2 --> 3)-2,4-di-O-benzoyl-6-O-levulinoyl-D galactopyranosyl trichloroacetimidate was coupled with 2-(trimethylsilyl)ethyl (2 acetamido-2-deoxy- 3-O-benzyl-6-O-p-methoxyphenyl-beta-D-glucopyranosyl)-(1 --> 3)-(2,4,6-tri-O-benzyl-beta-D-galactopyranosyl)-(1 --> 4)-2,3,6-tri-O-benzyl-beta D-glucopyranoside to give the suitably protected pentasaccharide which, upon selective removal of the p-methoxyphenyl and/or levulinoyl groups at C-6 of the GlcNAc and the terminal Gal residues, successive O-sulfation(s) and deprotection, afforded the desired three sulfated IV3NeuAcnLcOse4 derivatives. Acceptor specificity of the synthetic IV3NeuAcnLcOse4 probes for a human alpha-(1 --> 3) fucosyltransferase (Fuc-TVII) was examined to study the biosynthetic pathway of L selectin ligand. Only the 6-sulfated derivative at C-6 of GlcNAc was recognized by Fuc-TVII to give 6-O-sulfo sialyl LeX. PMID- 11028777 TI - Total synthesis of alpha-galactosyl cerebroside. AB - A highly convergent synthetic approach was developed to obtain alpha-galactosyl cerebroside O-(alpha-D-galactopyranosyl)-2-hexacosylamino-D-ribo-1,3,4-octa decantriol, which has previously been demonstrated to have immunostimulatory activity. Known 4,6-O-benzylidene galactose was the starting material for both the required alpha-galactosyl and the phytosphingosine building blocks O-(2,3-di O-benzyl-4,6-O-benzylidene-D-galactopyranosyl) trichloroacetimidate (4) and 2-O methanesulfonyl-D-arabino-1,2,3,4-octadecantetrol (5). The key step of the synthetic strategy is the highly regio- and stereoselective O-galactosylation of 1,3,4-O-unprotected phytosphingosine acceptor 5 using known 4 as donor. The total synthesis required only 11 synthetic steps starting from galactose. PMID- 11028778 TI - Interaction of saccharides with rare earth metal ions: synthesis and characterisation of Pr(III)- and Nd(III)-saccharide complexes. AB - A number of complexes are synthesised with Pr(III) and Nd(III) employing the mono and disaccharides, D-glucose, D-galactose, D-mannose, D-arabinose, D-ribose, D xylose, maltose and lactose. The complexes are characterised by various spectral and analytical techniques, and based on these data, appropriate structures were assigned for all complexes. PMID- 11028780 TI - Anomeric effects in non-carbohydrate compounds: conformational differences between the oxazolidine rings of a cis-fused bicyclic system. AB - Tris(hydroxymethyl)aminomethane (Tris) can react with benzaldehyde (1:2 molar ratio) to produce cis-2,8-diphenyl-5-hydroxymethyl-1-aza-3,7 dioxabicyclo[3.3.0]octa ne, the structure of which has been confirmed by nuclear magnetic resonance spectroscopy and X-ray crystallography. The crystal structure showed that both oxazolidine rings A and B are puckered in opposite directions. Ring A exists in an E3 envelope form with 0-3 noticeably down (0.65 A) the plane of the remaining atoms, whereas ring B adopts the 7E envelope conformation with the 0-7 atom displaced up from the mean reference plane by 0.70 A. Comparison of bond angles and bond distances showed that both oxazolidine rings A and B exhibit cross endo-anomeric effects resulting from electron delocalization over the bond sequence O-3-C-2-N-1-C-8-O-7. PMID- 11028779 TI - Preparation and structure determination of two sugar amino acids via corresponding hydantoin derivatives. AB - (4R)-2,3-O-Isopropylidene-methylspiro[4,6-dideoxy-alpha-L-lyxo+ ++-hexopyranosid 4,5'-imidazolidin]-2',4'-dione and (4R)-2,3-O-isopropylidene-methylspiro[4,6 dideoxy-beta-D-ribo-h exopyranosid-4,5'-imidazolidin]-2',4'-dione were prepared under various reaction conditions starting from methyl 6-deoxy-2,3-O isopropylidene-alpha-L-lyxo-hexopyranosid-4-++ +ulose. Corresponding alpha-amino acids methyl (4R)-4-amino-4-C-carboxy-4,6-dideoxy-alpha-L-lyxo-hexopyranosid e and methyl (4R)-4-amino-4-C-carboxy-4,6-dideoxy-beta-D-ribo-hexopyranoside were obtained from the above hydantoins by selective acid hydrolysis of the isopropylidene group, followed by basic hydrolysis of the hydantoin ring. The crystal structures of both hydantoin derivatives are also presented. PMID- 11028781 TI - The crystal structure of the 1:1 complex of beta-cyclodextrin with trans-cinnamic acid. AB - The crystal structure of the 1:1 complex of beta-cyclodextrin (cyclomaltoheptaose) with trans-cinnamic acid was studied by X-ray diffraction. Two beta-cyclodextrin molecules related by a twofold crystal axis form dimers in the hydrophobic cavity of which, two guest molecules are entirely buried. The complex crystallizes in the monoclinic C2 space group with channel-type molecular packing. The oxygen atoms of the carboxylate group of the trans-cinnamic acid molecule form strong hydrogen bonds with two water molecules lying in the interdimeric space of the hydrophobic channel. PMID- 11028782 TI - Acid-catalyzed hydrolysis of hesperidin at elevated temperatures. AB - Dilute sulfuric acid was used as a catalyst for hydrolysis of hesperidin suspensions in water at temperatures ranging from 25 to 180 degrees C. Significant acceleration of the reaction was observed at 120 degrees C and higher temperatures. This increase in the rate of hydrolysis can be attributed to increased solubilization of hesperidin in water at higher temperatures. Partial hydrolysis of hesperidin at 140 degrees C was used for the preparations of hesperetin-7-glucoside, which has a value in the synthesis of dihydrochalcone sweeteners. Simple separation of hesperetin and hesperetin-7-glucoside by extraction with dry acetone or lower alcohols has been developed. PMID- 11028783 TI - A convergent synthesis of trisaccharides with alpha-Neu5Ac-(2 --> 3)-beta-D-gal (1 --> 4)-beta-D-GlcNAc and alpha-Neu5Ac-(2 --> 3)-beta-D-gal-(1 --> 3)-alpha-D GalNAc sequences. AB - The syntheses of three trisaccharides: alpha-Neu5Ac-(2 --> 3)-beta-D-Gal-(1 --> 4)-beta-D-GlcNAc --> OMe, alpha-Neu5Ac-(2 --> 3)-beta-D-Gal6SO3Na-(1 --> 4)-beta D-GlcNAc --> OMe, and alpha-Neu5Ac-(2 --> 3)-beta-D-Gal-(1 --> 3)-alpha-D-GalNAc -> OBn were accomplished by using either methyl (phenyl 5-acetamido-4,7,8,9-tetra O-acetyl-3,5-dideoxy-2-thio-beta-D-glycero-D-g alacto-2-nonulopyranoside)onate or methyl (phenyl N-acetyl-5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-thio beta-D-gl ycero-D-galacto-2-nonulopyranoside)onate as the sialyl donor. The N,N diacetylamino sialyl donor appears to be more reactive than its parent acetamido sugar when allowed to react with an disaccharide acceptor under the same glycosylation conditions. The trisaccharides, as well as the intermediate products, were fully characterized by 2D DQF 1H-1H COSY and 2D ROESY spectroscopy. PMID- 11028784 TI - The phase transformations in starch during gelatinisation: a liquid crystalline approach. AB - The analogy between starch and a chiral side-chain polymeric liquid crystal is examined in relation to the processes involved during gelatinisation. There are three important parameters for characterisation of the molecular phase behaviour of the amylopectin: the lamellar order parameter (psi), the orientational order parameter of the amylopectin double helices (phi), and the helicity of the sample (h, the helix/coil ratio, a measure of the helix-coil transition of the double helices). The coupling between the double helices and the backbone through the flexible spacers is affected dramatically by the water content and it is this factor which dictates the particular phase adopted by the amylopectin inside the starch granule as a function of temperature. SAXS, WAXS and 13C CP/MAS NMR are used to examine these phenomena in excess water. Furthermore, previous experimental evidence pertaining to the limiting water case is reviewed with respect to this new theoretical framework. PMID- 11028785 TI - Characterisation of the high-molecular weight fructan isolated from garlic (Allium sativum L.). AB - A high molecular weight fructan was isolated from garlic and the structure determined by enzymatic, chemical and spectroscopic (NMR) methods. It was found that the garlic fructan belongs to the neokestose family. It has a (2 --> 1) linked beta-D-Fruf backbone with (2 --> 6)-linked beta-D-Fruf side chains. A structural model was postulated for a degree of polymerisation of about 58. This model was substantiated using an endo-inulinase purified from Aspergillus ficuum and by 1H and 13C NMR spectroscopy. PMID- 11028786 TI - Characterization of arabinose and ferulic acid rich pectic polysaccharides and hemicelluloses from sugar beet pulp. AB - Pectic polysaccharides were extracted from sugar beet pulp to yield fractions representing homogalacturonans, rhamnogalacturonans, arabinans and relatively small amounts of glucomannans and xyloglucans. The homogalacturonans had an apparent molecular weight of 21 kDa and contained relatively high amounts of methyl esters and relatively low amounts of acetyl groups as compared with the ramified 'hairy' regions. Three populations which originated from the ramified 'hairy' regions of pectin were distinguished. Two of these were rhamnogalacturonans with high apparent molecular weights of 1300 and 120 kDa, respectively. These populations had a high Ara and ferulic acid content. Despite the high neutral sugar content, these rhamnogalacturonans strongly bound to a DEAE column. The third population which originated from the ramified 'hairy' regions was a neutral population, which did not interact with the DEAE column and had a low apparent molecular weight and a high Ara and ferulic acid content. The arabinan side-chains of the rhamnogalacturonans were heavily branched in all populations. Enzymatic degradation of the xyloglucans showed similarities with apple xyloglucans with respect to the substitution with Fuc and Gal. PMID- 11028787 TI - Oxidative cross-linking of pectic polysaccharides from sugar beet pulp. AB - Oxidative cross-linking of three beet pectin extracts with hydrogen peroxide/peroxidase resulted in an increase in viscosity at low concentrations and in the formation of a gel at higher concentrations. Gels were formed using concentrations of 1.5% for an autoclave preparation and one obtained by an acid extraction and of 3% for a second autoclaved extract. It was shown that in the autoclave extracts only rhamnogalacturonans and possibly the arabinans participated in the cross-linking reaction. Cross-linking of the autoclave extracts with ammonium persulfate resulted in a decrease in reduced viscosity and molecular weight, although ferulic acid dehydrodimers were formed. Treatment of the acid extracted pectin with ammonium persulfate gave a slow increase in viscosity and the formation of a high-molecular-weight population was observed. For both oxidative systems, the 8-5 dehydrodimer was predominant after cross linking. PMID- 11028788 TI - A wide diversity of sulfated polysaccharides are synthesized by different species of marine sponges. AB - Sulfated polysaccharides were extracted from four species of marine sponges by exhaustive papain digestion. These compounds were purified by anion-exchange and gel-filtration chromatography. Analysis of the purified polysaccharides revealed a species-specific variation in their chemical composition and also in their molecular masses. In the species Aplysina fulva we found a sulfated glucan with a glycogen-like structure. The other three species contained sulfated polysaccharides with variable proportions of galactose, fucose, arabinose and hexuronic acid and also with different degrees of sulfation. Although the complex nature of these polysaccharides did not allow complete structure determination, we detected the occurrence of 4-sulfated residues of fucose and arabinose in the species Dysidea fragilis. The biological role of these sulfated polysaccharides requires further investigation. They may be involved in the species-specific aggregation of sponge cells and/or in the structural integrity of sponge, resembling the proteoglycans of mammalian connective tissues. PMID- 11028789 TI - Crystal structures of heptakis(2,6-di-O-tert butyldimethylsilyl)cyclomaltoheptaose, heptakis(2-O-methyl-3,6-di-O-tert butyldimethylsilyl)cyclomaltoheptaose and heptakis(2-O-methyl)cyclomaltoheptaose. AB - Crystal structures of heptakis(2,6-di-O-tert butyldimethylsilyl)cyclomaltoheptaose, heptakis(2-O-methyl-3,6-di-O-tert butyldimethylsilyl)cyclomaltohep taose and heptakis(2-O-methyl)cyclomaltoheptaose were determined from X-ray diffraction patterns obtained for single crystals of the title compounds grown from ethyl acetate and ethanol, respectively, as solvent. The crystal structures prove conclusively that quantitative migration of the tert-butyldimethylsilyl group from the 2-O- to the 3-O-position [D. Icheln, B. Gehrcke, Y. Piprek, P. Mischnick, W.A. Konig, M.A. Dessoy, A.F. Morel, Carbohydr. Res., 280 (1996) 237-250] was achieved during methylation of heptakis(2,6-di-O-tert-butyldimethylsilyl)cyclomaltoheptaose by iodomethane sodium hydride. PMID- 11028790 TI - Structure of an O-acetylated acidic O-specific polysaccharide of Proteus vulgaris O46. AB - An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O46 and studied by chemical methods (O deacetylation, sugar and methylation analyses, partial solvolysis) and 1H and 13C NMR spectroscopy. Solvolysis of the O-deacetylated polysaccharide with trifluoromethanesulfonic acid resulted in a alpha-D-GlcpNAc-(1 --> 3)-D-GlcA disaccharide that demonstrated the usefulness of this reagent for selective cleavage of heteropolysaccharides. The following structure for the polysaccharide was established: --> 4)-alpha-D-Glcp6Ac(1 --> 3)-beta-D-GlcpA4Ac-(1 --> 3)-alpha D-GlcpNAc-(1 --> 3)-beta-D-GlcpA4Ac-(1 --> where the degree of O-acetylation is approximately 65% at position 6 of Glc and 80-95% at position 4 of GlcA residues. PMID- 11028791 TI - Observations on the crystallization of oligogalacturonates. AB - Oligogalacturonates were produced by the limited enzymic hydrolysis of polygalacturonic acid and purified by ion-exchange chromatography. The fractions obtained were of limited polydispersity, determined by analytical ion-exchange chromatography. Oligomers with an average degree of polymerization of 10-15 were readily crystallized from aqueous salt solutions at neutral pH as single crystals. Crystal morphology of the salts examined, Na+, K+ and Ca2+ were characteristic of the salt. The wide-angle X-ray diffraction patterns obtained for the sodium salt were consistent with published fibre diffraction data of this salt form. PMID- 11028792 TI - Molecular and crystal structure of galactinol dihydrate [1-O-(alpha-D galactopyranosyl)-myo-inositol dihydrate]. AB - The crystal structure of galactinol dihydrate has been determined by X-ray diffraction. The crystal belongs to the orthorhombic system, space group P2(1)2(1)2, a = 15.898(6), b = 19.357(5), c = 5.104(4) A, and Z = 4. The structure was refined to R = 0.044 for 1818 observed structure amplitudes. The primary hydroxyl group exhibits twofold orientational disorder. The linkage conformation is close to those of alpha-(1 --> 4) linkages in methyl alpha maltotrioside tetrahydrate and erlose trihydrate. Although there is no interring hydrogen bond in galactinol, an indirect interring hydrogen bond including a water molecule is present. The observed conformation is additionally stabilized by the indirect interring hydrogen bond. The global minimum in the relaxed residue energy map based on the MM3(92) force-field is close to the observed conformation in the crystal structure. All hydroxyl, ring and water oxygen atoms are involved in a complex three-dimensional hydrogen-bonding network. PMID- 11028793 TI - Physical activity for elderly persons with neurological impairment: a review. AB - The paper reviews studies conducted on physical activities and exercise in elderly persons with neurological impairment due to stroke, Guillain-Barre syndrome, Parkinson's disease, multiple sclerosis or post-polio syndrome. The paper concludes: (i) it is not possible at present to draw conclusions regarding persons with Guillain-Barre syndrome and Parkinson's disease; (ii) individuals with multiple sclerosis and post-polio syndrome benefit from physical activity, but all studies have so far been conducted on those under 65 years of age, and its effect on elderly persons with these diseases is not known; (iii) exercise and customary activities (walking and swimming) should be encouraged and promoted in elderly persons after stroke. PMID- 11028794 TI - Quality of life assessment: its integration in rehabilitation care through a model of daily living. AB - To integrate quality of life assessment with rehabilitation care, some correspondence is required between the concepts of quality of life and of rehabilitation. A notion of quality of life is presented in which quality is conceived as degree of goodness, and life as daily living. Rehabilitation is considered both a process of adaptation and assistance to that process. These notions of quality of life and of rehabilitation can together be operationalized through a model of daily living. An individual's appraisal of his own situation in relation to adaptation can be explained, assuming a hierarchy of internal standards. Explaining appraisal by others requires external standards. Both types of appraisal are important grounds for decisions regarding assistance. In addition, general ideas on justification of rehabilitation as assistance may influence such decisions. The model integrates both objective and subjective appraisal and ideas on justification into rehabilitation, thereby offering opportunities for theoretical underpinning of the practice. PMID- 11028795 TI - Important factors for physical activity among elderly patients one year after an acute myocardial infarction. AB - The aim of this study was to identify and describe the factors of importance for elderly (> or =65 years) patients in being physically active one year after acute myocardial infarction. Forty-three consecutive elderly patients with a recent myocardial infarction were randomized either to a supervised outpatient-group training programme, 50 min three times a week for 3 months, or to a control group. An independent observer interviewed the patients 12 months after randomization in order to elucidate the factors that motivated the patients into being physically active. Both groups were identical at the start. The patients in the training group stated that the programme had made them more self-confident regarding physical activities and this seems to be an important factor for continuing to be physically active. Body mass index, age, gender and support from a physically active partner were of minor importance compared to the training programme or earlier experience of regular physical activity. PMID- 11028796 TI - Back muscle fatigue in healthy men and women studied by electromyography spectral parameters and subjective ratings. AB - To obtain reference data for future studies of patients with low back pain, back muscle fatigue was studied by surface electromyography at L1 and L5 lumbar levels in 55 healthy subjects exerting 80% of maximal voluntary contraction of the back extensors in a sitting position. Reference data were the initial value and rate of decrease (slope) of the median frequency during the contraction. The aim was also to study the effects of contraction time, gender differences, electrode locations and correlations with torque, age and subjective ratings. Initial median frequency was 52 Hz +/- 7.5, with no difference between electrode locations; steeper slopes were found at L5 level (-0.44%/s +/- 0.25) than at L1 ( 0.36%/s +/- 0.26). No right-left differences and no gender differences were found for these parameters. A correlation was observed between slope and initial median frequency, higher for men (r approximately -0.7) than for women (r approximately 0.5). Intersubject coefficient of variation for the slope was smallest for the longest (45 seconds) recording time (60-70%), but still much higher than for the initial median frequency (14%). The torque and the subjective ratings of fatigue showed no correlation with the electromyography variables. We conclude that the same reference values can be used for men and women. Owing to the large intersubject range of the slope, the clinical use of this variable may, however, be impeded. PMID- 11028797 TI - Correlations between joint and spinal mobility, spinal sagittal configuration, segmental mobility, segmental pain, symptoms and disabilities in female homecare personnel. AB - The aim of a study comprising 607 women working as homecare personnel was to investigate general spinal, joint and segmental mobility, different symptoms (pain and strain) and their relation to various aspects of disability. Joint mobility (mainly peripheral) was estimated using the "Beighton" score and spinal posture and mobility were measured by kyphometer. Passive segmental mobility and pain provocation were estimated manually. Pain intensity and strain during work and leisure were estimated using visual analogue scales for defined anatomical regions. Disability was rated using defined items and two indices. The 7-day prevalence of low back pain was 48%. Peripheral joint mobility, spinal sagittal posture and thoracic sagittal mobility showed low correlations with disability. Lumbar sagittal hypomobility was associated with higher disability. Manually estimated segmental mobility and segmental pain provocation of L4-L5 and L5-S1 correlated with disability; hypo- and hypermobility or positive pain provocation tests at these levels showed higher disability than normal mobility and negative pain provocation tests, respectively. Cluster analysis revealed that the combination of positive pain provocation tests and low lumbar sagittal mobility was associated with particularly high disability levels. In conclusion, positive pain provocation tests were clearly associated with high disability levels. PMID- 11028798 TI - Shoulder rotational strength, movement, pain and joint tenderness as indicators of upper-extremity activity limitation in moderate rheumatoid arthritis. AB - In this study 32 women were investigated in order to elucidate how shoulder rotational muscular strength and upper-extremity impairments are associated with activity limitation in moderate rheumatoid arthritis. A regression analysis was carried out to determine whether these variables could indicate the outcomes of a shoulder-arm disability questionnaire (SDQ) comprising three parts, plus parts of the Health Assessment Questionnaire, the Functional Status Questionnaire and the Sickness Impact Profile. Shoulder-arm and wrist movements were moderate-to-good (r = 0.53, p < 0.01 and r = 0.58, p < 0.01, respectively) in relation to isometric internal rotational strength. The relationship between isokinetic concentric and eccentric internal rotational strength was moderate-to-good (r = 0.59, p < 0.01). Isokinetic eccentric internal rotation strength, shoulder-arm movement, joint tenderness and pain variables together indicated 25-61% (adjusted R2) of the variation in SDQ. Eccentric strength had the highest adjusted R2 (41%) in relation to SDQ 1, covering mainly personal hygiene. Shoulder rotational strength did not indicate the more general instruments. Thus, hand and elbow impairments also are probably important in explaining activity limitations. PMID- 11028799 TI - Validity and responsiveness of the rivermead mobility index in stroke patients. AB - The Rivermead Mobility Index is used to measure mobility in patients with head injury or stroke. The purpose of the study was to examine construct validity, predictive validity, and the responsiveness of the Rivermead Mobility Index in stroke patients. Thirty-eight stroke inpatients participated in the study. The Rivermead Mobility Index, the Barthel Index, and the Berg Balance Scale were administered at admission to the rehabilitation ward and at discharge. The results showed that the Rivermead Mobility Index fulfilled the Guttman scaling criteria (coefficients of reproducibility > 0.9, coefficients of scalability > 0.7). The Rivermead Mobility Index scores were highly correlated with the Barthel Index scores (Spearman rs > 0.6) and the Berg Balance Scale scores (Spearman rs > = 0.8, all ps < 0.001). The Rivermead Mobility Index score at admission was closely correlated with the Barthel Index score at discharge (Spearman r = 0.77, p < 0.001). About 76% (29) of the subjects improved by more than 3 Rivermead Mobility Index points (median = 5) during their stay. The relationship between the change in score of the Rivermead Mobility Index and the Barthel Index was fair (Spearman r = 0.6, p < 0.001). These results indicate that the Rivermead Mobility Index is valid and sensitive to change over time. It is therefore a useful scale for the assessment of mobility in stroke patients. PMID- 11028800 TI - Validity of functional independence measure scores. PMID- 11028801 TI - Absence of latent Epstein-Barr virus in thymic epithelial tumors as demonstrated by Epstein-Barr-encoded RNA(EBER) in situ hybridization. AB - BACKGROUND: Several studies have established that Epstein-Barr virus (EBV) is associated with lympho-proliferative disorders such as Burkitt's lymphoma and Hodgkin's disease. EBV is also present in undifferentiated nasopharyngeal carcinomas and in tumors of similar morphology (lymphoepithelioma-like carcinomas) arising in a variety of organs, predominantly in stomach, salivary gland and thymus. As reports of EBV-positive thymic epithelial tumors (TET) have been divergent and as different methods have been used to detect EBV, the aim of this study was to investigate the possible role of EBV in TET of Danish patients. MATERIAL AND METHODS: Archival material of 157 cases of TET (105 thymomas and 52 thymic carcinomas, including 4 lymphoepithelioma-like thymic carcinomas (LELTC)) was analyzed for EBV by applying a sensitive and specific method for detecting latently EBV-infected cells (in situ hybridization for EBV-encoded RNA (EBER)). RESULTS: All investigated cases were EBER negative. CONCLUSIONS: EBV does not seem to be implicated in the pathogenesis of TET. However, a review of the literature showed that 28% of LELTC were EBER ISH positive. As they occurred in young people (mean 18 years), at an age when the patients were susceptible to infection by EBV, it is suggested that EBV merely acts as an innocent bystander. PMID- 11028802 TI - Legionella species of different human prevalence induce different rates of apoptosis in human monocytic cells. AB - Legionella species of different human prevalence were examined with respect to induction of apoptosis in the human monocytic cell line Mono Mac 6 (MM6). L. pneumophila serogroup 1 (Pontiac), L. pneumophila serogroup 1 (Philadelphia-1), L. longbeachae serogroup 1, L. gormanii, L. micdadei and L. steigerwaltii were used to infect MM6 cells. Subsequent induction of apoptosis was investigated by enzyme-linked immunosorbent assay (ELISA), gel electrophoresis of cellular DNA extracts, and staining of cells with the DNA dye 4', 6-diamidino-2-phenylindole (DAPI). Additionally, the concomitant occurrence of infection and apoptosis was demonstrated by a combination of immunohistochemistry with nuclear DAPI counterstaining. Induction of apoptosis in MM6 cells by a given species of the genus Legionella correlates with their human prevalence rather than with their ability to multiply within this human monocytic cell line. Furthermore, we found that initiation of apoptosis of Mono Mac 6 cells was dependent on direct adherence of the pathogenic bacteria to the host cell and was triggered by extracellular bacteria. PMID- 11028803 TI - Reduced phagocytic activity of polymorphonuclear leukocytes in alpha(1,3) fucosyltransferase VII-deficient mice. AB - Deficiencies in adhesion molecules or their counter-receptors in humans may have severe consequences as exemplified by leukocyte adhesion deficiency (LAD) I or II syndromes. Because such diseases occur with great rarity, animal models are valuable for studying the role of particular adhesion molecules and their natural ligands in immunity. We studied selected immune parameters and general health in mice with a defect in the sialyl-Lewis X antigen (selectin ligand) caused by disruption of the gene encoding alpha(1,3)fucosyltransferase VII (Fuc-TVII). Leukocytes from Fuc-TVII -/- and control mice were tested for adherence to cellophane membranes or polymer particles in vivo and phagocytic activity in vitro. While no difference in adherence was found, the number of neutrophil granulocytes in exudate induced by intraperitoneal injection of polymer beads was reduced in knock-out mice. Moreover, the phagocytic activity in Fuc-TVII -/- mice was significantly reduced. These animals have splenomegaly due to increased hematopoiesis and reduced weight but do not exhibit clinical signs of immunodeficiency. In conclusion, the lack of Fuc-TVII activity leads to several morphological and functional abnormalities without an impact on survival rate. PMID- 11028805 TI - Prenatal malformed lumbar vertebral corpora in trisomies 21, 18, and 13, evaluated radiographically and histologically. AB - The aim of the present study was to compare, both radiographically and histologically, malformed vertebral lumbar corpora in trisomies 21, 18 and 13 with earlier reported normal corporal development in the axial lumbar region. Axial skeletons of human fetuses (GA 15-22 wk) derived from therapeutically induced abortion were investigated in connection with requested autopsy. The number of lumbar vertebral corpora examined for each genotype was as follows: 20 from trisomy 21, 10 from trisomy 18, and 10 from trisomy 13. After radiography in frontal, lateral and axial projections, the individual vertebral corpora were decalcified and horizontally embedded in paraffin. The blocks were serially sectioned and stained with toluidine blue and alcian blue/van Gieson. The radiographic characteristics of the vertebral corpora varied from an almost normal appearance of the corporal bone to complete clefting of the bony corpora. Histological examination showed accumulations of cartilage centrally, in some cases associated with amorphous material. Pronounced metachromatic differences were observed in the cartilaginous ground substance. The study showed identical phenotypic characteristics in the corpora from trisomy 21, trisomy 18, and trisomy 13. It is characteristic of all three genotypes that there are central anomalies, corresponding to the location of the notochord in normal corpora, and marked regional differences in metachromasia in the ground substance of the cartilage. PMID- 11028804 TI - Reduced ammonium chloride haemolysis time enhances the number of isolated functional rabbit polymorphonuclear neutrophils. AB - In the present study we compared seven different methods for isolating rabbit polymorphonuclear neutrophils (PMNs) with a view to assessing viability, lymphocyte contamination and isolation yield. The two methods offering the best isolation yield and functional PMNs were retained. Leukocyte-containing plasma fraction was obtained after erythrocyte sedimentation with dextran. First, PMNs were isolated from this fraction, using hypotonic ammonium chloride haemolysis followed by Histopaque density gradient centrifugation (Method-A). Second, PMNs were obtained from leukocyte-containing plasma after centrifugation on two Percoll layers (Method-B). These processes resulted in a high cellular yield: 2.66x10(6)+/-0.22 PMNs per ml of blood (Method-A) and 1.87x10(6)+/-0.37 PMNs per ml of blood (Method-B). In both cases the PMNs isolated were of high purity and viability. In comparison, when using the standard techniques for rabbit - consisting of ammonium chloride haemolysis taking at least four times as long- fewer PMNs were isolated. The PMNs isolated by Method-A and -B were able to generate a high amount of reactive oxygen species (ROS) after stimulation with phorbol 12-myristate 13-acetate (PMA). These methods to separate PMNs are recommended for in vitro studies. PMID- 11028806 TI - Strain differentiation in Bacteroides fragilis by ribotyping and computer assisted gel analysis. AB - It is important to distinguish between Bacteroides fragilis strains as their virulence may vary and as B. fragilis seems to be a heterogeneous species. The aim of our study was to evaluate ribotyping for differentiation of 46 strains of B. fragilis and for assessment of strain heterogeneity within and between the two DNA-DNA homology strain groups established in this species. Twenty-seven strains belonged to Johnson's DNA homology group I and eight to group II. Eleven strains had not been assigned to any group (NI group). DNA from all strains was cut with BglI, EcoRI and HindIII. Restriction fragment length polymorphisms were investigated using a non-radioactive digoxigenin-labelled cDNA probe transcribed from Escherichia coli 16S+23S rRNA. Ribotyping with BglI was most discriminatory, revealing a total of 26 different patterns by visual inspection of gels. EcoRI followed with 20 patterns and HindIII with 13 patterns. The gels from ribotyping were processed using the Dendron computer-assisted program. Strain clusters established using Dendron were not always in agreement with homology-based strain groups. Strains of the NI group fell into both homology groups. Ribotyping, as it is based on a relatively small portion of the genome, is useful for strain distinction in epidemiological studies with B. fragilis, whereas DNA-DNA homology, using the entire genome, is more reliable for taxonomy. The Dendron computer-assisted program, which enabled objective assessment of multiple banding patterns, increased the reliability of ribotyping. PMID- 11028807 TI - Modes of adherence of Helicobacter pylori to gastric surface epithelium in gastroduodenal disease: a possible sequence of events leading to internalisation. AB - We have investigated various modes of adherence of Helicobacter pylori to the human gastric epithelium, using transmission electron microscopy, in biopsies from nine patients with peptic ulcer disease and from four patients with chronic active gastritis. H. pylori was demonstrated in abundance in all cases within the surface mucous layer. In all ulcer- and in one out of four gastritis patients H. pylori was shown in close proximity to the gastric epithelium, with concurrent alterations in the configuration of microvilli and the apical cytoplasmic region of gastric cells. Previously described modes of H. pylori adherence were confirmed, such as loose attachment with fibrillar-like strands, firm attachment with pedestal formation, invasion in the intercellular spaces, and invagination with "cup" formation. Moreover, in many cases a fusion between the bacterial outer layer and gastric cell membranes was evident. In four cases (31%; three with active and one with past ulcer disease) viable H. pylori was found in the cytoplasm of gastric mucous cells. Our results support the hypothesis that the different modes of adherence of H. pylori represent a stepwise, possibly sequential, process which in a significant number of cases leads to internalisation of the organism. The invariable occurrence of adhesion and more frequent internalisation of H. pylori in ulcer patients may suggest a link with the pathogenesis of peptic ulcer disease. PMID- 11028808 TI - Argyrophilic nucleolar organizer regions in Helicobacter pylori-associated gastric lesions. AB - Three hundred and fifty biopsies from patients undergoing upper gastrointestinal endoscopy were studied for histopathological changes, H. pylori infection and argyrophilic nucleolar organizer region (AgNOR) counts. Histopathological examination revealed normal gastric mucosa in 10 (2.85%), gastritis in 254 (72.56%), intestinal metaplasia in 12 (4.0%), dysplasia in 13 (3.7%) and adenocarcinoma in 61 (17.4%). The mean (SD) AgNOR count was 1.66 (0.20) in normal, 2.43 (0.64) in gastritis, 3.09 (0.52) in intestinal metaplasia, 4.17 (0.31) in dysplasia, and 6.57 (0.98) in carcinoma. A statistically significant difference was observed between the AgNOR count of normal gastric mucosa and gastritis (p<0.001), gastritis and dysplasia (p<0.001), and dysplasia and adenocarcinoma (p<0.001). A statistically significant increase in mean AgNOR count was found with increase in H. pylori density in gastric biopsies (p<0.001) with gastritis. No significant difference was observed between mean AgNOR count of intestinal and diffuse type carcinomas. The AgNOR count in gastric biopsies with adenocarcinoma and H. pylori infection was 7.03 (0.85) as compared to 6.89 (0.73) in gastric biopsies with evidence of adenocarcinoma but without H. pylori infection. The difference was not statistically significant. The findings support the role of H. pylori as a promoting agent in gastric carcinogenesis by stimulating gastric epithelial cell proliferation at the stage of chronic inflammation, thereby making the cells more susceptible to endogenous or exogenous carcinogenic agents. PMID- 11028809 TI - Blood culture bottles for transportation and recovery of anaerobic bacteria from non-blood samples. AB - Using bacterial suspensions as simulated non-blood specimens, the capacity of three different BacT/ Alert blood culture bottles for the transportation and recovery of anaerobic bacteria with different sensitivity to air was evaluated. To better assess the performance of the BacT/Alert bottles, three other liquid media specially designed for anaerobes were included in the study. Attention was paid to recovery rates in relation to species, initial bacterial concentration, and time needed for detection. Of the BacT/Alert blood culture bottles, the anaerobic FAN bottle yielded the highest recovery rates, but its performance was limited compared with chopped meat broth in tubes. This broth allowed detection of all the tested species within 48 h. Since collection and transportation of anaerobic bacteria are of major importance for a reliable culture result, improvements are necessary. PMID- 11028810 TI - Human T-lymphocyte virus type I (HTLV-I)-induced myeloneuropathy in rats: oligodendrocytes undergo apoptosis in the presence of HTLV-I. AB - To investigate the pathogenetic role of human T-lymphocyte virus type I (HTLV-I) in central nervous system disease, a rat model for HTLV-I-associated myelopathy/tropical spastic paraparesis, designated as HAM rat disease, was examined with regard to chronological neuropathology, from early asymptomatic phase to late disease. In the thoracic spinal cord of rats with HTLV-I infection, the first event was the appearance of apoptosis of oligodendrocytes beginning at 7 months after induced infection, thereafter followed by the appearance of white matter degeneration, increase of macrophages/activated microglia and of gemistocytic astrocytes at 12, 15 and 20 months, respectively. In the spinal cord, HTLV-I provirus DNA was evident as early as 4 months after the infection, and HTLV-I pX and the tumor necrosis factor (TNF)-alpha messages began to be expressed at age 7 months, just before or at the same time as the appearance of apoptotic cells. Collective evidence suggests that the apoptotic death of oligodendrocytes, which may be induced either directly by the local expression of HTLV-I or indirectly by TNF-alpha, through the transactive function of p40Tax, is the major cause of chronic progressive myeloneuropathy in Wistar-King-Aptekman Hokudai rats with HTLV-I infection. PMID- 11028811 TI - The role of cytokeratins 20 and 7 and estrogen receptor analysis in separation of metastatic lobular carcinoma of the breast and metastatic signet ring cell carcinoma of the gastrointestinal tract. AB - Metastatic signet ring cell carcinomas of unknown primary site can represent a clinical problem. Gastrointestinal signet ring cell carcinomas and invasive lobular carcinomas of the breast are the most common sources of these metastases. Immunohistochemical algorithms have been successfully used in the search for the unknown primary adenocarcinomas. In the present study a series of primary invasive lobular breast carcinomas (79 cases) and their metastases and a series of gastrointestinal signet ring cell carcinomas (22 primary and 13 metastases) were stained with monoclonal antibodies for cytokeratin (CK) 20 and CK7 and for estrogen receptors (ER). The staining was evaluated as negative (no staining), focally (less than 10% of the tumor cells stained) or diffusely positive. All the primary and metastatic gastrointestinal signet ring cell carcinomas proved to be CK20 positive, while only 2/79 (3%) of the primary and 1/21 metastatic lobular carcinomas (5%) stained positively for this CK. None of the gastrointestinal carcinomas and the majority of the lobular carcinomas expressed ER. The majority of the tumors were CK7+. Using CK20 alone, 33 of 34 metastases could be properly classified as gastrointestinal (CK20+) or mammary (CK20-). ER identified 31/34 of breast cancer metastases. By combining the results of CK20 and ER staining all the metastases could be properly classified as the CK20+/ER- pattern identified all the gastrointestinal tumors. PMID- 11028812 TI - Interleukin 4 and interferon-gamma expression of the dermal infiltrate in patients with erythroderma and mycosis fungoides. An immuno-histochemical study. AB - BACKGROUND: Erythroderma, or generalized erythema of the skin, may result from different causes. At present it is unclear whether the underlying patho mechanisms that lead to erythroderma are identical or different depending on the original disease. The aim of this study was to investigate the dermal cytokine profile in different types of erythroderma and mycosis fungoides. METHODS: Snap frozen skin biopsy specimens from 33 patients with erythroderma were studied. Thirteen had idiopathic erythroderma, 7 erythrodermic atopic dermatitis, 5 Sezary syndrome and 8 had erythroderma from miscellaneous causes. We also studied 6 patients with mycosis fungoides (5 plaques and 1 tumor) and 5 healthy non-atopic volunteers. The biopsies were immunohistochemically stained for interleukin 4 (IL 4) and interferon gamma (IFN-gamma). All positive cells for IL-4 and IFN-gamma in the dermis were counted and the number of positive cells was calculated per mm2. IL-4/IFN-gamma ratio was calculated for each biopsy. RESULTS: The patients with idiopathic erythroderma, atopic dermatitis and miscellaneous erythroderma, all showed more IFN-gamma-positive cells than IL-4-positive cells in the dermis. The median IL-4/ IFN-gamma ratio for these three groups was 0.6, 0.9 and 0.45, respectively. These differences were not statistically significant. All patients with Sezary syndrome however, showed more IL-4-positive cells than IFN-gamma positive cells. The median IL-4/IFN-gamma ratio was 1.8, which is significantly higher than in the other groups p<0.05). In mycosis fungoides roughly the same number of cells expressed IL-4 and IFN-gamma. The median IL-4/IFN-gamma ratio was 1.0, which is significantly lower than in Sezary syndrome (p<0.05). CONCLUSIONS: The dermal infiltrate in patients with Sezary syndrome mainly shows a T-helper 2 (Th2) cytokine profile, this in contrast to T-helper 1 (Th1) cytokine profile in benign reactive erythroderma. This indicates that although a relative uniform clinical picture of erythroderma is obvious, a different patho-mechanisms may be underlying. PMID- 11028813 TI - Expression of VCAM-1, ICAM-1, E-selectin, and P-selectin on endothelium in situ in patients with erythroderma, mycosis fungoides and atopic dermatitis. AB - BACKGROUND: Erythroderma may result from different causes. At present it is unclear whether the patho-mechanisms that lead to these different types of erythroderma are identical or different. Adhesion molecules and their ligands play a major role in endothelial-leukocyte interactions, which affect the binding, transmigration and infiltration of lymphocytes and mononuclear cells during inflammation, injury, or immunological stimulation. The aim of this study was to investigate the adhesion molecule expression on endothelial cells in erythroderma in situ. METHODS: Snap-frozen skin biopsy specimens from 23 patients with erythroderma were studied. Eight had idiopathic erythroderma, 5 erythrodermic atopic dermatitis, 4 Sezary syndrome and 6 had erythroderma from miscellaneous causes. As a control we studied skin specimens from 10 patients with mycosis fungoides, 5 patients with atopic dermatitis and 5 healthy non atopic volunteers. To determine adhesion molecule expression on endothelial cells in situ, sections were immuno-histochemically double stained with biotinylated Ulex Europaeus agglutinin 1 as a pan-endothelial cell marker, and for the adhesion molecules VCAM-1, ICAM-1, E-, and P-selectin. All double- and single stained blood vessels in the dermis were counted. RESULTS: Mean endothelial expression in erythroderma was as follows: VCAM-1 51.4%, ICAM-1 70.1%, E-selectin 43.5%, and P-selectin 52.6%. There was no statistical difference between different groups of erythroderma. Mean expression of all adhesion molecules tested, was in Sezary syndrome higher than in mycosis fungoides albeit not significant. In erythrodermic atopic dermatitis only VCAM-1 expression was significantly higher than in lesional skin of atopic dermatitis. No differences were observed in expression of the other three adhesion molecules. CONCLUSIONS: There is no difference regarding adhesion molecule expression on endothelial cells between different types of erythroderma. PMID- 11028814 TI - In vitro characteristics of neonatal hemangioma endothelial cells: similarities and differences between normal neonatal and fetal endothelial cells. AB - BACKGROUND: Increased angiogenesis and eventual involution are major characteristics of neonatal hemangiomas. The mechanism to explain this transition is not completely understood. METHODS: To determine the nature of these changes, endothelial cells were isolated from eight hemangiomas and the growth characteristics and morphology of these cells were compared to cells isolated from normal fetal and neonatal skin. Three cells lines were further characterized by analyzing protein expression with immunohistochemistry and FACS analysis. RESULTS: Hemangioma endothelial cells converted to a spindle-shaped morphology similar to that of fetal endothelial cells whereas neonatal endothelial cells maintained their characteristic epithelioid morphology. While neonatal, hemangioma and fetal endothelial cells continued to express platelet-endothelial cell adhesion molecule-1 (PECAM-1) and von Willebrand factor (vWf), hemangioma and fetal cells expressed both proteins at a lower level and in a distribution distinct from normal neonatal endothelial cells. Neonatal endothelial cells continued to express epithelial specific Type IV collagen, while hemangioma and fetal endothelial cells produced interstitial Type I collagen. CONCLUSIONS: Both cell morphology and protein expression of neonatal hemangioma endothelial cells were more characteristic of embryonic microvascular endothelial cells than that of postembryonic cells demonstrating a similarity in these two cell types and suggesting a dysfunction in the normal growth and maturation of endothelial cells in this tumor. PMID- 11028815 TI - Solitary fibrous tumors are immunophenotypically distinct from mesothelioma(s). AB - INTRODUCTION: Solitary fibrous tumor is a soft tissue tissue tumor of unknown histogenesis. Based upon histologic similarities and CD-34 expression, it has been suggested that these neoplasms bear some relationship to mesothelioma, and may represent its extra-pleural equivalent. METHODS: In order to further investigate this possible relationship, we examined a series of five dermal and five extra-cutaneous solitary fibrous tumors with antibodies directed against the mesothelial markers calretinin and HBME-1. RESULTS: All the lesions failed to stain with the antibodies tested. This suggests that despite some similar histologic and immunophenotypic features, these lesions are not immunophenotypically identical. Mesotheliomas are CD-34(+), calretinin(+), HBME 1(+), while solitary fibrous tumors are CD-34(+), calretinin(-), HBME-1(-). CONCLUSIONS: The histogenesis of solitary fibrous tumor remains elusive. It is unlikely that tumor location or tumor de-differentiation accounts for the dichotomous staining properties, as these neoplasms show a similarly benign histologic appearance regardless of location. PMID- 11028816 TI - Rippled-pattern sebaceous trichoblastoma. AB - BACKGROUND: There is a large spectrum trichoblastoma; of which, several histologic variants have been described including a rippled-pattern variant. Three cases of rippled-pattern trichoblastoma are described which also exhibited definitive foci of sebaceous differentiation. METHODS: Three cases were retrieved from the archives of the Dermatopathology Laboratory at the University of California Irvine (Orange, CA, USA). All specimens were stained with hematoxylin and eosin (H&E). In addition, sections were submitted for immunohistochemical studies with epithelial membrane antigen (EMA). RESULTS: All three biopsies were composed of well-circumscribed multiple variously sized tumor lobules present in the upper to deep dermis comprised of with rounded or slightly elongated basaloid cells with scant eosinophilic cytoplasm. The lobules were separated by a slightly hyalinized stroma. The unique finding present in all three specimens was a peculiar arrangement of the basaloid cells in linear rows parallel to one another. This gave the tumors a rippled pattern similar to the palisading of nuclei of Verocay bodies seen in schwannomas. In addition all three biopsies showed definite sebaceous differentiation. CONCLUSIONS: Three additional cases of rippled-pattern trichoblastoma are presented. All three were located on the scalp and showed additional features of foci of sebaceous differentiation. No associations with Muir-Torre Syndrome were found in these patients. Because this appears to be a distinct variant within the large spectrum of trichoblastoma, the term rippled-pattern sebaceous trichoblastoma is suggested. PMID- 11028817 TI - A rippled-pattern trichoblastoma: an immunohistochemical study. AB - BACKGROUND: Since the first description by Hashimoto et al., there have been only a few case reports of rippled-pattern tricogenic tumor. In addition, there are no reports on detailed immunohistochemical analyses of this rare neoplasm. We describe here an additional case of rippled-pattern trichogenic tumor with a special reference to its immunohistochemical features. METHODS: A nodule arising on the occipital area of a 62-year-old Japanese woman was histologically and immunohistochemically investigated. RESULTS: Histopathologically, the lesion contained various-sized lobular nests, which consisted of oval to elliptical shaped basaloid cells without any atypia and were embedded in the collagenous stroma. Some elongated basaloid cells were arranged in a palisading fashion forming parallel rows of epithelial ribbons in a rippled-pattern. Cytokeratin (CK) immunohistochemistry showed constant expressions of CK1/5/ 10/14, CK5/8, CK14 and CK7, and focal expressions of CK17 and CK19 in the basaloid cells, suggesting a keratin phenotypical similarity to the cells in small nodular type trichoblastoma. CONCLUSIONS: The present tumor is a variant of trichoblastoma, and considered to be in close association with the outer root sheath and/ or follicular germinative cells. PMID- 11028818 TI - A fatal case of pancreatic panniculitis presenting in a young patient with systemic lupus. AB - BACKGROUND: Subcutaneous fat necrosis associated with pancreatic disease is a rare event. The clinical cutaneous findings are non-specific erythematous nodules with central softening located predominantly on the lower extremities. The histopathologic features of these lesions are very characteristic and diagnostic. METHODS: We present an unusual case of pancreatic panniculitis associated with lupus pancreatitis in a 21-year-old African American female. The patient presented with lower extremity skin nodules, arthralgia, and serositis prior to the diagnosis of systemic lupus and pancreatitis. The skin lesions progressed despite normalization of serum pancreatic enzymes. Following femoral vein catheterization for renal dialysis, she developed a large indurated area over the left lower quadrant, flank, groin, and upper thigh measuring 25 cm. She was treated with repeated debridement, tissue grafts, and hyperbaric oxygen because of a clinical suspicion of necrotizing fasciitis. RESULTS: Examination of skin biopsies and debrided tissue revealed the pathognomonic features of pancreatic panniculitis without any evidence of necrotizing fasciitis. Organisms were not detected by tissue examination or microbiologic cultures. CONCLUSIONS: This case illustrates the potential role of vascular trauma in the pathogenesis of pancreatic panniculitis. PMID- 11028819 TI - Intra-epidermal and intra-dermal sebocrine adenoma with cystic degeneration and hemorrhage. AB - BACKGROUND: The ducts of eccrine glands may give rise to intra-epidermal, confluent epithelial and intra-dermal adenomas known as hidroacanthoma simplex, eccrine poroma, and dermal duct tumor, respectively. An apocrine and sebaceous counterpart of the eccrine poroma has been described by several authors as adnexal, poroma-like adenoma with apocrine and sebaceous differentiation or sebocrine adenoma. METHODS: Using clinical history and routine histologic techniques, we describe a new lesion with features similar to sebocrine adenoma but representing the intra-epidermal and intra-dermal counterparts with cystic degeneration and hemorrhage. Briefly, an 84-year-old female presented with a 6 mm dark tan papule on the neck that clinically appeared as an unusual macular seborrheic keratosis with underlying hemorrhage. RESULTS: Histopathological examination showed a benign dermal cystic appendage tumor with pale polygonal cells, occasional non-keratinizing ducts, sebaceous differentiation and central hemorrhage with fibrin deposits. Serial sections did not reveal any epidermal connection. However, epithelioid cells with large nuclei in an intra-epidermal pagetoid pattern were focally seen. CONCLUSION: These findings represent a new cystic, hemorrhagic variant of sebocrine adenoma. PMID- 11028820 TI - Langerhans' cell histiocytosis arising at the site of basal cell carcinoma excision. AB - Langerhans' cell histiocytosis (LCH) may be associated with a wide range of neoplastic disorders. However, the combination of a LCH and a basal cell carcinoma (BCC) of the skin still represents a highly unusual condition. In this publication, we report the case of a 48-year-old woman who developed a localized LCH involving the area of a previously excised BCC. Although the exact pathogenesis of this peculiar association remains yet to be elucidated, the hypothesis of an exaggerated Langerhans' cell reaction in response to the underlying BCC appears to be attractive. PMID- 11028821 TI - E-cadherin expression in basal cell carcinoma: association with local invasiveness but not with metastatic inefficiency. PMID- 11028822 TI - Melanoma and extravascular migratory metastasis. PMID- 11028823 TI - Fibromyalgia-symptom or diagnosis: a definition of the position. AB - According to the American College of Rheumatology the diagnosis of fibromyalgia is based on criteria for the classification of fibromyalgia consisting entirely of clinical signs and symptoms. For diagnostic reasons autonomic disturbances and mental features have to be considered. The distinction between fibromyalgia (tender points) and myofascial pain syndrome (trigger points) is essential. Internal and neurological disorders as a primary cause of fibromyalgia have to be excluded. The etiology and pathogenesis of fibromyalgia still remain uncertain. The myopathological patterns in fibromyalgia are non-specific: type II fiber atrophy, an increase of lipid droplets, a slight proliferation of mitochondria, and a slightly elevated incidence of ragged red fibers. Initial reports on some allelic abnormalities in the serotonin system seem to highlight the important role of serotonin already presumed earlier. Significantly high levels of substance P in the cerebrospinal fluid of FM patients additionally support the impact of these neurotransmitters on both nociceptive and antinociceptive mechanisms. PMID- 11028824 TI - Neuroendocrine and hormonal perturbations and relations to the serotonergic system in fibromyalgia patients. AB - The symptomatology of the fibromyalgia syndrome (FMS) often resembles an alteration in central nervous set points at least in three systems. The patients suffer under chronic pain in the region of the locomotor system, presumably reflecting a disturbed central processing of pain. Anxiety and depression often characterizes the clinical picture. Almost all of the hormonal feedback mechanisms controlled by the hypothalamus are altered. Characteristic for FMS patients are the elevated basal values of ACTH, follicle-stimulating hormone (FSH), and cortisol as well as lowered basal values of insulin-like growth factor 1 (IGF-1, somatomedin C), free triiodothyronine (FT3), and oestrogen. In FMS patients, the systemic administration of the relevant releasing hormones of corticotropin-releasing hormone (CRH), growth hormone-releasing hormone (GHRH), thyreotropin-releasing hormone (TRH), and luteinizing hormone-releasing hormone (LHRH) leads to increased secretion of ACTH and prolactin, whereas the degree to which TSH can be stimulated is reduced. The stimulation of the hypophysis with LHRH in female FMS patients during their follicular phase results in a significantly reduced LH response. All in all, the typical alterations in set points of hormonal regulation that are typical for FMS patients can be explained as a primary stress activation of hypothalamic CRH neurons caused by the chronic pain. In addition to the stimulation of pituitary ACTH secretion, CRH activates somatostatin on the hypothalamic level, which in turn inhibits the release of GH and TSH on the hypophyseal level. The lowered oestrogen levels could be accounted for both via an inhibitory effect of the CRH on the hypothalamic release of LHRH or via a direct CRH-mediated inhibition of the FSH-stimulated oestrogen production in the ovary. Serotonin (5HT), precursors like tryptophan (5HTP), drugs which release 5HT or act directly on 5HT receptors stimulate HPA axis, indicating a stimulatory serotonergic influence on HPA axis function. Therefore activation of the HPA axis may reflect an elevated serotonergic tonus in the central nervous system of FMS patients. PMID- 11028825 TI - Neuroimaging of chronic pain: phantom limb and musculoskeletal pain. AB - New developments in brain imaging lead to a better understanding of cortical and subcortical processes involved in pain perception and the establishment of chronic pain. For different forms of chronic pain long-term changes in cortical structures have been described. In patients with phantom limb pain and back pain alterations in the somatotopic organization of the primary somatosensory (SI) could be observed. The amount of this reorganization is correlated with the subjective pain rating. These changes, which are based on processes of neuronal plasticity, can partially be reversed by analgesic interventions. For the investigation of cortical processes concerning reorganization, EEG and MEG methods are most suitable because of their high temporal and spatial resolution. In conclusion, these findings open a new way for therapeutic interventions to prevent the development of chronic pain. PMID- 11028826 TI - Activity-dependent changes in the pain matrix. AB - Repetitive synaptic excitation or the application of L-glutamate into the vicinity of multireceptive neurons in the dorsal horn of the spinal cord and corresponding structures of the trigeminal nucleus increases neuronal excitability, which is then reflected by an expansion of the receptive field (Fig. 1). Similar alterations of the receptive field of neurons have been observed in various other brain regions. The receptive fields of multireceptive neurons also expand their size following mechanical, chemical, inflammatory or nerve injuries. Since these multireceptive neurons are activated by converging non-nociceptive and nociceptive afferents an increased excitability of these neurons may also be the mechanism by which pain refers to distant somatic and visceral structures (Fig. 2). The increase in neuronal excitability is mediated to a great extent by the co-activation of glutamate receptors and receptors for substance P, a neuropeptide long thought to have a role in pain perception. There is evidence from recent research that this facilitatory effect on glutamatergic synaptic transmission involves membrane receptor phosphorylation, and enhances activity-dependent gene expression (Fig. 3). In order to investigate the time dependent processing of ongoing afferent noxious stimulation in the central nervous system we recently employed the quantitative autoradiographic 14C-2 deoxyglucose technique in a model of chronic monoarthritic pain in the rat. A synopsis of these most recent experimental data and results from previous electrophysiological in vivo and in vitro studies suggests that dorsal horn neurons and probably also other neurons in pain-related structures become spontaneously active and can maintain their activity without further noxious peripheral input. PMID- 11028827 TI - Neurobiological concepts of fibromyalgia--the possible role of descending spinal tracts. AB - In the spinal cord, long descending pathways are known to exist which modulate pain sensations by either inhibiting or facilitating the discharges of spinal nociceptive neurones. In this article, the hypothesis is discussed that the pain of fibromyalgia may be due to a dysfunction of these pain-modulating pathways. Theoretically, two kinds of disturbance could lead to pain, namely reduced activity in the pain-inhibiting (antinociceptive) system or increased activity in the pain-facilitating (pronociceptive) pathways. Data from animal experiments show that interruption of the dorsal descending systems leads to hyperactivity of spinal nociceptive neurones, namely increase in background activity, lowering in stimulation threshold, and increase in response magnitude to noxious stimuli. The responses of the neurones to input from nociceptors in deep tissues were more strongly inhibited by the descending pathways than were responses to input from cutaneous nociceptors. Collectively, the findings indicate that the dorsal descending systems are tonicly active and have a particularly strong inhibitory action on neurones that mediate pain from deep tissues. If these systems operate in a similar way also in patients, an impairment of their function is likely to lead to 1. spontaneous deep pain (because of an increased background activity in nociceptive neurones supplying deep tissues), 2. tenderness of deep tissues (because of a lowered mechanical threshold of the same neurones), and 3. hyperalgesia of deep tissues (because of increased neuronal responses to noxious stimuli). These changes will affect large areas of the body because the descending inhibitory systems have widespread terminations in the spinal cord. Thus, a dysfunction of the descending inhibitory pathways could mimick to a large extent the pain of fibromyalgia. PMID- 11028828 TI - The role of psychosocial factors in fibromyalgia syndrome. AB - OBJECTIVE: The main objective of this review was to evaluate the role of psychosocial factors in the development of fibromyalgia syndrome. METHOD: Review of the literature concerning the influence of psychosocial factors. RESULTS: In fibromyalgia syndrome psychosocial factors are relevant at different etiological levels. They can be classified into predisposing, triggering and stabilising/"chronifying" factors. CONCLUSION: Due to the increasing knowledge about the influence of psychosocial factors for the development of fibromyalgia, the biomedical model has to be expanded to a biopsychosocial model. The biopsychosocial concept has an impact on the therapeutic approach. Strong evidence for the model is provided by the good results of interdisciplinary treatment studies. PMID- 11028829 TI - Present state of medication therapy in fibromyalgia syndrome. AB - For the treatment of primary fibromyalgia syndrome (FMS) the low dose application of tri- and tetracyclic antidepressive drugs was often studied. Up to now from all those drugs the effects of amitriptyline (AMI) are best documented. Because of its sedative properties it doesn't only influence pain but also improves the often disturbed sleep. Its use in patients with FMS is limited by the occurrence of side effects and the lack of response in a substantial number of patients. Serotonin reuptake inhibitors alone seem to be of little value. Nevertheless there is evidence that they may improve pain in combination with other antidepressive agents. Regarding pain moclobemide a reversible inhibitor of monoamine oxidase seems to be inferior to AMI. In controlled studies corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs) also failed to improve FMS. The combination of NSAIDs with benzodiazepines gave inconsistent results. Although often used, we have only small information about the effectiveness of opioids. No beneficial effect could be attributed to the muscle relaxant chlormezanone. In conclusion, although only about 1/3 of the patients respond, AMI remains the drug of first choice in the conventional medication treatment of FMS. PMID- 11028830 TI - Preclinical and clinical pharmacology of the 5-HT3 receptor antagonists. AB - 5-HT3-receptor antagonists are potent and highly selective competitive inhibitors of the 5-HT3-receptor with negligible affinity for other receptors. They are rapidly absorbed and penetrate the blood-brain barrier easily. 5-HT3-receptor antagonists are metabolized by diverse subtypes of the cytochrome P450-system, metabolites are excreted mainly in urine. Half-lifes in healthy subjects vary from 3-4 hours (ondansetron, granisetron) to 7-10 hours (tropisetron, hydrodolasetron). 5-HT3-receptor antagonists do not modify any aspect of normal behaviour in animals or induce remarkable changes of physiological functions in healthy subjects. They are well tolerated over wide dose ranges, most common side effects in clinical use are headache and obstipation. Clinical efficacy was first established in chemotherapy-induced emesis. In this indication, 5-HT3-receptor antagonists set a new standard regarding efficacy and tolerability. Further established indications are radiotherapy-induced and post-operative emesis. Antiemetic efficacy results from a simultaneous action at peripheral and central 5-HT3-receptors. Other peripheral actions include reduction of secretion and diarrhea caused by increased intestinal serotonin content (e.g. in carcinoid syndrome), a limited antiarrhythmic activity and a reduction of experimentally induced pain. CNS effects comprise anxiolysis, attenuation of age-associated memory impairment, reduction of alcohol consumption in moderate alcohol abuse and an antipsychotic effect in patients with parkinson psychosis. In migraine, 5-HT3 receptor antagonists show moderate efficacy, as well. Repeatedly demonstrated efficacy of 5-HT3-receptor antagonists in patients suffering from fibromyalgia raises the question for the mechanism of action involved. Ligand binding at the 5 HT3-receptor causes manifold effects on other neurotransmitter and neuropeptide systems. In particular, 5-HT3-receptor antagonists diminish serotonin-induced release of substance P from C-fibers and prevent unmasking of NK2-receptors in the presence of serotonin. These observations possibly provide an approach for the causal explanation of favourable treatment results with 5-HT3-receptor antagonists in fibromyalgia. PMID- 11028831 TI - Effect of tropisetron on circulating catecholamines and other putative biochemical markers in serum of patients with fibromyalgia. AB - OBJECTIVE: The aim of the study was to assess the influence of the 5HT3-receptor antagonist tropisetron on circulating catecholamines as biochemical markers of the activity of the sympathoadrenal system in fibromyalgia. Moreover, serum concentrations of serotonin, somatomedin C, oxytocin, calcitonin-gene-related peptide, calcitonin and cholecystokinin were assayed as putative markers in pain related disorders like primary fibromyalgia. METHODS: In 96 patients, who met the ACR classification criteria for fibromyalgia, and in 20 sex and age matched controls concentrations of dopamine, noradrenaline, adrenaline, serotonin and tropisetron were assayed in serum by HPLC with electrochemical detection. All other transmitters were determined by ELISA. RESULTS: There was with the exception of tropisetron, calcitonin and dopamine, no correlation between doses of tropisetron 5, 10, 15 mg respectively and significant changes in circulating transmitters or other transmitters as putative biochemicals markers in primary fibromyalgia. Regarding the prediction of pain reduction to tropisetron, patients with elevated dopamine and/or reduced plasma 5-HT concentrations tended to show a higher response rate. CONCLUSION: Despite these partly disappointing results another prospective pilot study with selected patients vs. age and sex matched controls, double blind and with comparison of other 5HT3-receptor antagonists e.g. dolasetron and granisetron e.g. after i.v. bolus injection is suggested. Still the data obtained in this preliminary paper provide some evidence regarding the present discussion on subgroups of patients with primary fibromyalgia. PMID- 11028833 TI - Oral treatment of fibromyalgia with tropisetron given over 28 days: influence on functional and vegetative symptoms, psychometric parameters and pain. AB - The 5-HT3 receptor antagonists are a novel therapy for patients suffering from fibromyalgia, although the optimal duration of treatment is still unclear. The objective of this phase II study was to evaluate whether prolonging treatment with tropisetron to 4 weeks is tolerable and correlated with an improved clinical benefit. Thirty female patients with fibromyalgia received oral tropisetron (5 mg) daily for 28 days in an open-label fashion. Treatment resulted in significantly decreased pain as measured by visual analog scale (VAS), with a mean reduction of 59.7% and an absolute median change of -25.0 from baseline to day 28 (p<0.0001). A similar, significant reduction of 55.7% and absolute median change of -31.0 was observed in the painscore (p<0.0001). The response rate with patients showing a > or = 35% reduction in individual pain scores was 72.4% at day 28. The pressure tolerance of tender-points was slightly increased at the end of the treatment period. In addition, significant improvements were observed in the State-Trait-Anxiety-Inventory (STAI), scales of von Zerssen (Bf-S) and Beck Depression Index (BDI). Functional symptoms were compared with the results from a 10-day, randomized, double-blind phase III study of tropisetron in 418 fibromyalgia patients. In both studies several functional symptoms such as sleep disturbances and dizziness improved significantly (p<0.05). In the 28 days study, the number and extent of improvement in functional symptoms was increased compared with the shorter trial. Tolerability and safety of tropisetron was good, and typically for 5-HT3-receptor antagonists, gastrointestinal symptoms and headache were the most frequently reported events. In conclusion, 28 days treatment of fibromyalgia patients with 5 mg tropisetron resulted in significant pain reduction, which was most pronounced after 10 days with a further reduction up to day 28. Psychometric tests showed significant improvements in depression and anxiety state scores, while functional symptoms improved with extended tropisetron treatment. PMID- 11028832 TI - Efficacy and tolerability of tropisetron in primary fibromyalgia--a highly selective and competitive 5-HT3 receptor antagonist. German Fibromyalgia Study Group. AB - OBJECTIVE: Based on a potential role for serotonin receptors in fibromyalgia, we investigated the efficacy and tolerability of treatment with tropisetron, a highly selective, competitive inhibitor of the 5-HT3 receptor. METHODS: In this prospective, multicenter, double-blind, parallel-group, dose-finding study, 418 patients suffering from primary fibromyalgia (ACR criteria) were randomly assigned to receive either placebo, 5 mg, 10 mg or 15 mg tropisetron once daily, respectively. The duration of treatment was 10 days. The clinical response was measured by changes in pain-score, visual analog scale (VAS), and the number of painful tender-points. RESULTS: Treatment with 5 mg tropisetron resulted in a significantly higher response rate (39.2%) when compared with placebo (26.2%) (p=0.033). The absolute reduction in pain-score was -13.5% for 5 mg tropisetron, 13.0% for 10 mg tropisetron, and -6.3% for placebo (p<0.05). The effects of 15 mg tropisetron were similar to placebo, thus suggesting a bell-shaped dose-response curve. Compared with placebo, treatment with 5 mg tropisetron led to a significant improvement (p<0.05) in VAS, while a clear trend in terms of clinical benefit was seen with 10 mg tropisetron. The number of painful tender-points was also reduced significantly (p=0.002) in the 5 mg tropisetron group. Of interest, during the 12-month follow-up period, pain intensity of responders on 5 mg and 10 mg tropisetron was still markedly below baseline. The treatment was well tolerated, with gastro-intestinal complaints being the most frequently reported side effects, in keeping with the known safety profile for 5-HT3 receptor antagonists. CONCLUSIONS: This study demonstrates the efficacy of short-term treatment with 5 mg tropisetron once daily in primary fibromyalgia. Treatment was well tolerated and prolonged clinical benefits were seen. PMID- 11028834 TI - Results of the intravenous administration of tropisetron in fibromyalgia patients. AB - The observed effects on the symptoms of fibromyalgia of daily oral administration of 5 mg of the 5-HT3 receptor antagonist, tropisetron, for 10 days, could be maintained or exceeded with intravenous administration of only 2 mg of the formulation. Following a single i.v. injection of 2 mg tropisetron, a more rapid and profound reduction in pain was achieved than with 5 mg oral tropisetron per day. In individual cases, patients who had previously experienced no reduction in pain from 10 days of 5 mg oral tropisetron daily responded to i.v. therapy. A more favourable and persistent effect on pain, combined with a simultaneous significant improvement in various vegetative and functional symptoms was achieved with five days treatment with 2 mg tropisetron i.v. per day. The results outlined and the possibility for rapid improvements with drug treatment of fibromyalgia should be confirmed in randomised, placebo controlled trials. PMID- 11028835 TI - Do predictors exist for the therapeutic effect of 5-HT3 receptor antagonists in fibromyalgia? AB - From the findings outlined below, there are no reliable predictors of the therapeutic effect of the 5-HT3-receptor antagonists in fibromyalgia. Neither clinical change in pain and vegetative symptoms, nor alterations in biochemical parameters are appropriate predictors of response. The accompanying psychological changes in the form of depressive disorders appear to be somewhat predictive of decreased therapeutic effect, if such definitive statements can be applied to individual cases. If, following new trials, it becomes possible to judge the response of patients to therapy after 3-5 days treatment with 2 mg intravenous tropisetron then predictors will be unnecessary in practice. PMID- 11028836 TI - The use of 5-HT3 receptor antagonists in various rheumatic diseases--a clue to the mechanism of action of these agents in fibromyalgia? AB - In a pilot study, the action of the 5-HT3 receptor antagonist, tropisetron, on different types of local rheumatic pain and inflammatory effects was studied. With intra-articular injection of tropisetron, an improvement in inflammation and pain was obtained in inflammatory rheumatic diseases and activated osteoarthrosis. Also, the majority of patients with localized soft-tissue rheumatic diseases (periarthritis) demonstrated an obvious decrease in their pain following local infiltration of tropisetron. Chronic low back pain and cervical pain responded somewhat to i.v. treatment with tropisetron. The effect of the 5 HT3 receptor antagonists is probable primarily to limit the release of substance P, which acts as a pain and inflammatory mediator, and is itself released by the neurogenic inflammation that occurs after the binding of serotonin to its corresponding receptor. These results should be backed up with placebo controlled studies, which if confirmed, might imply that 5-HT3 receptor antagonists could supplement or replace the local administration of corticosteroids. PMID- 11028838 TI - Physical therapy in the treatment of fibromyalgia. AB - Fibromyalgia (FM) is a syndrome of unknown etiology characterized by chronic wide spread pain, increased tenderness to palpation and additional symptoms such as disturbed sleep, stiffness, fatigue and psychological distress. While medication mainly focus on pain reduction, physical therapy is aimed at disease consequences such as pain, fatigue, deconditioning, muscle weakness and sleep disturbances and other disease consequences. We systematically reviewed current treatment options in the treatment of fibromyalgia. Based on evidence from randomized controlled trials cardiovascular fitness training importantly improves cardiovascular fitness, both subjective and objective measures of pain as well as subjective energy and work capacity and physical and social activities. Based on anecdotal evidence or small observational studies physiotherapy may reduce overloading of the muscle system, improve postural fatigue and positioning, and condition weak muscles. Modalities and whole body cryotherapy may reduce localized as well as generalized pain in short term. Trigger point injection may reduce pain originating from concomitant trigger points in selected FM patient. Massage may reduce muscle tension and may be prescribed as a adjunct with other therapeutic interventions. Acupuncture may reduce pain and increase pain threshold. Biofeedback may positively influence subjective and objective disease measures. TENS may reduce localized musculoskeletal pain in fibromyalgia. While there seems to be no single best treatment option, physical therapy seem to reduce disease consequences. Accordingly a multidisciplinary approach combining these therapies in a well balanced program may be the most promising strategy and is currently recommended in the treatment of fibromyalgia. PMID- 11028837 TI - Treatment of chronic fatigue syndrome with 5-HT3 receptor antagonists- preliminary results. AB - OBJECTIVE: The serotonin system presumably is involved in the pathogenesis of chronic fatigue syndrome (CFS). Results from a few studies led to the hypothesis of a "postsynaptic hyperresponsiveness" in CFS. Therefore we intended to evaluate the efficacy of 5-HT3 receptor antagonists in the treatment of CFS. PATIENTS AND METHODS: 2 patient groups (10 patients each; CFS according to the CDC classification criteria) received either oral tropisetron (5 mg once daily) or oral ondansetron (2 x 8 mg daily), open-labelled. Treatment duration was 15 days. Treatment response was evaluated by visual analog scales (VAS) for fatigue and capability. RESULTS: 19 patients finished their respective study. In the tropisetron group 6/9 (VAS fatigue) and 7/9 (VAS capability) patients documented benefit, 8/10 rsp. 8/10 patients in the ondansetron group. The score changes (VAS before and after treatment) in case of response were more pronounced in the tropisetron group. The frequency of concomitant symptoms did not differ significantly in the treatment groups. The overall analysis of both studies showed a remarkable improvement (> or = 35%) of approximately one third of the patients in both VAS. Treatment was well tolerated. CONCLUSION: Our preliminary results encourage to perform placebo-controlled, double-blind studies to further evaluate the efficacy of 5-HT3 receptor antagonists in the treatment of CFS. PMID- 11028839 TI - The challenge of fibromyalgia: new approaches. PMID- 11028840 TI - Is it possible to offer evidence-based treatment for systemic vasculitis? AB - The evidence base for treatment of systemic vasculitis is reviewed in this article. Accurate diagnosis and evaluation of disease status is crucial to the appropriate management of these cases and disease status should be employed as a means of staging so that appropriate therapy can be offered tailored to the individual requirements of each case. Large vessel vasculitides are almost exclusively managed with corticosteroids. Small vessel vasculitis with internal organ involvement usually requires more aggressive immuno-suppression; by contrast, small vessel vasculitis without internal organ involvement rarely requires intervention. Despite our current efforts, the long-term morbidity of patients with systemic vasculitis remains high. We need better therapeutic strategies in order to limit or control disease rapidly and thereby prevent the accumulation of subsequent damage. Large-scale randomised trials comparing different therapeutic strategies offer the only hope of establishing a firmer evidence base for the optimal management of these diseases. PMID- 11028841 TI - In vitro migration of mononuclear cells towards synovial fluid and plasma from rheumatoid arthritis patients correlates to RANTES synovial fluid levels and to clinical pain parameters. AB - OBJECTIVE: To evaluate in vitro migration of mononuclear cells towards synovial fluid (SF) and plasma in relation to RANTES synovial fluid levels and clinical disease activity. METHODS: 31 RA patients with synovitis in one knee were included. Modified Boyden chamber technique was used to determine a migratory index defined as: 'In vitro migrating cells towards SF' divided by 'In vitro migrating cells towards plasma'. RANTES was quantified by ELISA. Disease activity was assessed by the swollen joint count, the Ritchie articular index (RAI), global assessment, pain on VAS, HAQ, ESR and CRP. RESULTS: A positive significant correlation was found between the migratory index and the RANTES levels in SF (r=0.48, p=0.006), the RAI (r=0.56, p=0.0001) and pain on VAS (r=0.43, p=0.04). The in vitro migration could be inhibited in 3 of 4 SF samples by neutralising antibodies towards RANTES (12-18%). CONCLUSION: The migratory index correlate to SF levels of RANTES and parameters for joint pain. PMID- 11028842 TI - Suppression of collagen-induced arthritis with histone H1. AB - Besides roles in nucleus mediating the condensation of DNA into chromatin, the involvement of histones in autoimmune diseases, hormone regulation, and killing leukemia cells has been reported. In order to investigate the functions of histones on an autoimmune disease, histone H1 was injected into collagen-induced arthritis (CIA) mice. A dramatic suppression of CIA by histone H1 was observed at a dose of 1 mg/kg bodyweight of mouse. In addition, the increased level of anti inflammatory cytokine IL-10 was detected in cultured splenocytes from the mouse treated with histone H1. These findings suggest that histone H1 suppresses the collagen-induced arthritis, possibly by increasing the level of IL-10 production. PMID- 11028843 TI - BPI-ANCA is found in reactive arthritis caused by Yersinia and Salmonella infection and recognise exclusively the C-terminal part of the BPI molecule. AB - OBJECTIVE: To examine the prevalence, binding sites and functional interactions of antineutrophil cytoplasmic autoantibodies (ANCA) against the bactericidal/permeability increasing protein (BPI) in reactive arthritis (ReA). METHODS: Sera were analysed for the occurrence of ANCA by indirect immunofluorescence microscopy (IIF) and ELISA. Binding sites were determined using BPI, lipopolysaccharid binding protein (LBP), and fusion proteins of both proteins in ELISA. In addition, the effect of antibodies on the antibiotic activity of BPI was examined. RESULTS: BPI-ANCA was found in patients with Yersinia- and Salmonella-triggered ReA and directed against the C-terminal portion of BPI. Goat anti BPI antibodies recognising this part inhibited the antibiotic activity of BPI in vitro. CONCLUSION: BPI-ANCA was associated with ReA triggered by Salmonella and Yersinia infection. Directed against the C-terminal part of BPI, it can potentially inhibit its antibiotic activity and might be useful to identify patients with infectious bowel disease prone to extraintestinal sequelae. PMID- 11028844 TI - Intensive treatment of rheumatoid arthritis reduces need for dialysis due to secondary amyloidosis. AB - We have analysed changes in the intensity of treatment of patients with rheumatoid arthritis (RA) and its influence on the number of patients with RA and secondary amyloidosis (SA) admitted to dialysis due to end-stage renal disease. The number of visits and patients at the rheumatological outpatient clinic because of RA, the type of medication used in 1988-1997, and the number of patients with RA and SA on dialysis from 1989 to 1999 were extracted from the data set of Kuopio University Hospital. The intensity of treatment could be evaluated as the treated number of patients with RA and the number of visits at the outpatient clinic. Both the numbers increased from 1988 to 1996, patients under treatment from 201 to 550 and visits from 1091 to 2198, respectively. In 1997, the number of patients still increased (n=637), although the number of visits started to decline (n=2054), partly due to better collaboration of health centres. A marked shift from use of only symptomatic treatment or one disease modifying antirheumatic drug (DMARD) to more common use of immunosuppressants and/or combinations of at least two DMARDs occurred in the five years from 1992 to 1997. In 1988, the figures for only nonsteroidal anti-inflammatory drugs or only glucocorticoids or one immunosuppressive drug were 24%, 8%, and 9%, and in 1997, 4%, 3%, and 22%, respectively. In 1997, 43% of the patients were treated with combinations of two or more DMARDs. The number of patients with RA on dialysis treatment and the number of new admissions each year due to SA decreased from 11 to 2 and from 5 to none, respectively. At the beginning of year 2000 there was only one patient with RA and SA on dialysis. PMID- 11028846 TI - Reliability and validity of a Swedish version of the Revised Leeds Disability Questionnaire for patients with ankylosing spondylitis. AB - The aim of the study was to investigate the reliability and the validity of a Swedish version of the Revised Leeds Disability Questionnaire (RLDQ). Forty-two patients, 36 men and 6 women, median age 52.5 years, median symptom duration 24 years, with spondylarthropathy were assessed with the RLDQ, three questions on the content of the questionnaire, and range-of-motion measures. The results indicated satisfactory test-retest stability and internal consistency, and only minor internal redundancy. RLDQ items were generally considered relevant and suggested additions were similar to items already included in the questionnaire. Patients with low disability scores tended to be older and have better joint mobility than those with higher scores. Physiotherapists observed and scored disability significantly higher than patients. Correlations between subscores of the RLDQ and range-of-motion measures were in the main weak. In conclusion the Swedish version of the RLDQ may be considered as reliable and valid. PMID- 11028845 TI - Pulmonary hypertension is associated with impaired exercise performance in patients with systemic sclerosis. AB - OBJECTIVE: The aim of this study was to evaluate the exercise tolerance by expired gas analysis during stress test in patients with Systemic Sclerosis (SSc). METHODS: Eighteen women (mean age 48.56+/-12.48 years) affected by SSc were studied. A complete echocardiographic examination including pulmonary artery systolic pressure estimation, pulmonary function tests, diffusion lung capacity for carbon monoxide (DLCO), and exercise test were performed. During exercise, breath-by-breath expired gas analysis was performed. RESULTS: Seven patients (39%) had baseline pulmonary systolic hypertension (group A) and 11 patients (61%) did not (group B). Six patients had reduced DLCO values. Both maximal oxygen consumption (VO2max) and anaerobic threshold (VO2AT) values were markedly decreased compared to the predicted values. Seven of 18 patients were unable to complete a maximal exercise (5 of whom affected by pulmonary systolic hypertension). Group A patients showed reduced VO2max, VO2AT, and O2 pulse compared with patients with group B patients (p=0.004, 0.017, and 0.013, respectively); VO2max, VO2AT and O2 pulse were significantly correlated to baseline pulmonary artery systolic pressure. CONCLUSIONS: An exercise intolerance in patients affected by SSc is present. Impairment of exercise performance is associated with pulmonary hypertension. PMID- 11028847 TI - MRI of cervical spine with flexion and extension used in patients with rheumatoid arthritis. AB - Patients with rheumatoid arthritis (RA) often have involvement of the cervical spine. The most common abnormality is atlanto-axial subluxation (AAS). The more serious vertical subluxation (VS) is thought to develop at a later stage. Direct cord compression may occur, but the symptoms may be vague and difficult to interpret. In addition to clinical follow up, RA patients undergo several conventional radiographs of the cervical spine, with addition of flexion and extension images. This, in spite of the fact that the cervical cord and soft tissue do not show. Magnetic resonance imaging (MRI), is the modality of choice to visualize soft tissue and the cervical medulla, but is rarely performed in the follow up of RA patients. Five patients with long-standing RA, episodes of neck pain, and known AAS were asked to volunteer for a MRI study of the cervical spine, consisting of sagittal T2 weighted images of the cervical spine during flexion and extension of the neck. Compared to clinical examinations and cervical radiographs, MRI gave valuable information not otherwise obtained. The importance of MRI with the neck in a flexed and extended position is stressed. This is possible to obtain within a conventional quadrature neck coil in many RA patients. PMID- 11028848 TI - Ultrasound evaluation of plantar fasciitis. AB - OBJECTIVE: To investigate the sonographic features of plantar fasciitis (PF). METHODS: High-resolution ultrasound was used to measure the thickness and echogenicity of the proximal plantar fascia and associated heel pad thickness for 102 consecutive patients with PF (unilateral: 81, bilateral: 21) and 33 control subjects. RESULTS: The mean thickness of the plantar fascia was greater on the symptomatic side for patients with bilateral and unilateral PF than on the asymptomatic side for patients with unilateral PF, and also control subjects (5.47+/-1.09, 5.61+/-1.19, 3.83+/-0.72, 3.19+/-0.43 mm, respectively, p<0.001). A substantial difference in thickness between the asymptomatic side of patients with unilateral PF and control subjects was also noted (p=0.001). The heel pad thickness was not show different between control subjects and patients with PF. The incidence of hypoechoic fascia was 68.3% (84/123). Other findings among the patients from our test group included intratendinous calcification (two cases), the presence of perifascial fluid (one case), atrophic heel pads (one case), and the partial rupture of plantar fascia (one case). CONCLUSION: Increased thickness and hypoechoic plantar fascia are consistent sonographic findings in patients exhibiting PF. These objective measurements can provide sufficient information for the physician to confirm an initial diagnosis of PF and assess individual treatment regimens. PMID- 11028849 TI - Arthritis as presenting symptom in silent adult coeliac disease. Two cases and review of the literature. AB - We report 2 cases of adult silent coeliac disease (CD) presenting with arthritis of a knee and a sacro-iliac joint, respectively. In both patients the arthritis was relieved on a gluten free diet. The literature on arthritis in adult CD is reviewed. PMID- 11028850 TI - Sudden death and Wegener's granulomatosis of the pituitary. AB - Involvement of brain parenchyma or meninges in ANCA-associated small-vessel vasculitis such as Wegener's granulomatosis (WG) is not uncommon. In contrast, involvement of the pituitary is exceedingly rare with only a few cases reported so far. The diagnosis is usually made on the basis of imaging techniques and abnormal pituitary function tests in the setting of active systemic vasculitis. However, histology-proven involvement of the pituitary by WG has not been reported so far. We report a case of WG with histology-proven granulomatous necrotizing inflammation of the pituitary and hypothalamo-pituitary stalk, disclosed at autopsy after the patient had died suddenly and unexpectedly in his sleep. In a setting of histology-proven WG, these findings were regarded as a pituitary manifestation of the disorder. A distinct cause of death could not be found, hence we speculate that hypothalamo-pituitary inflammation due to WG may have caused the sudden death in this patient. PMID- 11028851 TI - Sjogren's syndrome, cavitating lung disease and high sustained levels of antibodies to serine proteinase 3. AB - A case is described involving Sjogren's syndrome, high sustained levels of antibodies to serine proteinase 3, and cavitating lung disease. Possible diagnoses accounting for this unusual combination include a novel association of Sjogren's syndrome and Wegener's granulomatosis (suggested by the high and sustained levels of antibodies to serine proteinase 3) or a rare presentation of bronchiolitis obliterans organising pneumonia. Identification of the true nature of the patients illness facilitated more active management and a swift resolution of the clinical problem. PMID- 11028852 TI - False-positive result of systemic lupus erythematosus latex test induced by anti insulin antibody. PMID- 11028853 TI - Chronic fatigue syndrome and memory complaints. PMID- 11028854 TI - Intracutaneous injection of lysophosphatidylcholine induces skin inflammation and accumulation of leukocytes. AB - Various cell stimuli act through activation of phospholipase A2, which hydrolyses fatty acids from membrane phospholipids, resulting in the formation of fatty acids and lysophospholipids. One of the lysophospholipid classes, lysophosphatidylcholine, is a chemoattractant for monocytes and T-lymphocytes and induces the expression of adhesion molecules on cultured endothelial cells. The purpose of the present study was to determine whether lysophosphatidylcholine possesses proinflammatory properties in vivo. This was assessed clinically and histologically by intracutaneous injection of 200-800 nmol lysophosphatidylcholine in healthy volunteers. Lysophosphatidylcholine elicited a dose- and time-dependent local erythema and oedema. The erythema disappeared within 4 h, while the induration lasted for up to 48 h. HE-stained biopsies taken after 24 h showed a leukocytoclastic vasculitis in 2 of the 6 subjects. Microscopic examination of immunohistochemically stained biopsies taken 24 h after the injection showed a significant increase in the number of T-lymphocytes, monocytes and neutrophils, whereas the number of Langerhans' cells was unchanged after lysophosphatidylcholine injection. In addition, the number of intercellular cell adhesion molecule-1 and -3-positive cells was increased approximately 3-fold after injection of lysophosphatidylcholine. In conclusion, the phospholipase A2 hydrolysis product lysophosphatidylcholine can induce erythema, oedema, a mixed cellular infiltrate and the expression of adhesion molecules. PMID- 11028855 TI - Role of nerve growth factor in RANTES expression by keratinocytes. AB - A role of neurogenic inflammation induced by the neuropeptides and nerve growth factor (NGF) has been attributed to the pathogenesis of several cutaneous disorders such as psoriasis, wound healing and eczematous dermatitis. The underlying mechanisms of the inflammatory process induced by NGF are not clearly established. This study explored whether NGF influences the inflammatory process by inducing chemokines. The effects of NGF were investigated on induction of 2 important chemokines, interleukin-8 and RANTES, which are known to be upregulated in the keratinocytes of various inflammatory conditions. NGF significantly increased RANTES production by the keratinocytes (p < 0.001, 2-tailed Student's t test). Induction of RANTES expression in the keratinocytes by NGF provides further insight regarding the role of NGF-NGF receptor system in cutaneous inflammatory conditions. PMID- 11028856 TI - Expression of p21WAF1/CIP1, p53, bcl-2 and Ki-67 in venous leg ulcers with and without squamous cell carcinoma. AB - In order to gain understanding of the cellular mechanisms of malignant transformation in chronic venous leg ulcers, we analysed by immunohistochemistry the presence of p21, p53, bcl-2 and Ki-67 in ulcers with and without squamous cell carcinoma. The material consisted of 41 archived histopathological samples from 33 patients with chronic venous leg ulcers and 28 samples from 21 patients suffering from squamous cell carcinoma in chronic venous leg ulcers. All samples derived from the chronic ulcers were negative for p53, p21 and bcl-2. Cells positive for Ki-67 were seen in certain ulcer areas. In the cancers, 14 samples showed immunopositivity for p53 and 22 samples were positive for p21, with expression mainly in the suprabasal layer. Expression of p21 seemed independent of p53. None of the samples was bcl-2-positive. Ki-67 was positive with basal expression in 16/28 of the tumour samples, 15 of which also expressed p21. There was no perilesional p53 or p21 activity in the cancer samples. The results regarding perilesional expression are different from those reported in UV-induced squamous cell carcinomas and probably reflect a different carcinogen. PMID- 11028857 TI - Comparison of the sensitivities of the Buehler test and the guinea pig maximization test for predictive testing of contact allergy. AB - International test guidelines, such as the Organisation for Economic Cooperation and Development (OECD) guideline #406, recommend 2 guinea pig methods for testing of the contact allergenic potential of chemicals: the Guinea Pig Maximization Test (GPMT) and the Buehler test. Previous comparisons between the methods suggested that the Buehler test was less sensitive than the GPMT although modified Buehler test protocols were used. Parallel GPMT and Buehler tests were conducted according to OECD guideline #406 using a multiple-dose design and test results were analysed using a standard logistic dose-response model. To compare the sensitivity of the 2 test procedures the test conditions were kept identical and the following chemicals with a range of sensitization potentials were tested: chloraniline, chlorhexidine, eugenol, formaldehyde, mercaptobenzothiazole and neomycin sulphate. Formaldehyde and neomycin sulphate were strong sensitizers in both tests. Mercaptobenzothiazole, eugenol and chloraniline were all strong sensitizers in the GPMT, eugenol and mercaptobenzothiazole were negative in the Buehler test and equivocal results were obtained with chloraniline. Chlorhexidine was negative in the GPMT and equivocal responses were obtained with the Buehler test. Higher induction concentrations were needed to show allergenicity in the Buehler test and for some allergens the Buehler test protocol was not sensitive enough to demonstrate allergenic potential. PMID- 11028858 TI - In vivo study of skin mechanical properties in psoriasis vulgaris. AB - The aim of this study was to investigate the mechanical properties of the skin in psoriatic plaques before and after treatment with dithranol and in clinically uninvolved psoriatic skin in comparison with the skin of healthy controls. In total, 82 psoriatic plaques in 19 in-patients and 51 age-matched control subjects were studied with a non-invasive suction device (Cutometer). The plaques were characterized by statistically significant lower skin distensibility and elasticity, and a higher viscoelastic to elastic ratio compared with adjacent apparently normal skin. Delayed distension and hysteresis measured using a 2-mm diameter probe were decreased, but were increased when measured by a 8-mm diameter probe. These differences can be explained by the increased skin thickness in psoriasis and by the different aspects of skin mechanics determined by the 2 measuring probes. After treatment, the mechanical parameters of psoriatic plaques approached the values of adjacent control skin. The uninvolved volar forearm skin in psoriatic patients (2-mm diameter probe) showed statistically significant lower values of skin distensibility and higher values of viscoelasticity parameters compared with the skin of healthy control subjects. This suggests that the normal-appearing psoriatic skin is not entirely normal. The non-invasive method applied can be used for objective and quantitative evaluation of the effect of therapy in patients with psoriasis. PMID- 11028859 TI - Surgical removal of ulcer and lipodermatosclerosis followed by split-skin grafting (shave therapy) yields good long-term results in "non-healing" venous leg ulcers. AB - The purpose of this study was to evaluate the long-term effects of shave therapy in non-healing venous leg ulcers. Forty-one patients with 75 recalcitrant leg ulcers caused by primary deep vein incompetence or post-thrombotic syndrome were operated by shave therapy (removal of ulcer and surrounding lipodermatosclerosis with a Schink skin-grafting knife and covering of the wounds with meshed split thickness skin grafts). After an average follow-up period of 2 years and 5 months all patients were evaluated for long-term results. The healing rate of ulcers classified as non-healing was 67% (50 of 75 ulcers). The healing rate was 76% for ulcers associated with primary deep vein incompetence and 58% for ulcers associated with post-thrombotic syndrome (p = 0.08). Even in cases with recurrence (33%) these ulcers were strikingly reduced by 80-90% of their original size. Hypaesthesia was noticed in 38% of the transplanted areas. In "non-healing" venous leg ulcers due to deep venous insufficiency shave therapy yields favourable long-term results. Because it is only a symptomatic treatment which does not reduce the pathological refluxes, continuous compression of the lower leg is important. PMID- 11028860 TI - Dermatology life quality index: data from Danish inpatients and outpatients. AB - The aim of the present study was to provide data on the reliability and validity of a Danish translation of the Dermatology Life Quality Index (DLQI), a short measure of the impact of dermatological diseases on quality of life. The DLQI was administered to 200 outpatients and 100 hospitalized patients suffering from a range of dermatological diseases and to 100 sex- and age-matched healthy controls. Mean scores, internal consistency and test-retest reliability were comparable to the results reported for the original English version. Hospitalized patients reported greater impairment of disease-related quality of life than outpatients, and patients with atopic dermatitis and psoriasis exhibited greater scores than patients suffering from other dermatological diseases. Discriminant, construct and predictive validities of the Danish translation of the DLQI were satisfactory, as indicated by significant associations between DLQI scores and physician-rated disease severity, disease duration and the time patients were willing to spend each day on a hypothetical effective treatment. The results also suggest that the emphasis Danish patients place on various aspects of disability covered by the questionnaire is similar to that of English patients. In conclusion, the Danish translation of the DLQI showed satisfactory reliability and the preliminary results indicate that this version is a valid measure, which can be used in both research and clinical settings. PMID- 11028861 TI - Increased coexpression of eotaxin and interleukin 5 in bullous pemphigoid. AB - While the presence of eosinophils in the skin lesions of bullous pemphigoid is well documented, the chemotactic factors responsible for eosinophil recruitment into the tissue still remain to be defined. In this study, eotaxin and interleukin-5 (IL-5) concentrations were determined in the blister fluid and sera of patients with bullous pemphigoid (acute and remission phase, n=6) in comparison with normal healthy controls (n=6) using the enzyme-linked immunosorbent assay (ELISA) technique. Eotaxin and IL-5 levels were increased in the blister fluid compared with the acute and remission phase sera, as well as compared with the sera of normal controls. In addition, immunoreactivity for eotaxin was predominantly found in the inflammatory cell infiltrate of lesional bullous pemphigoid biopsy specimens. In conclusion, the data provide evidence that co-operation of eotaxin and IL-5 may play an essential role in activating and recruiting eosinophils, which ultimately contribute to the tissue damage in bullous pemphigoid. PMID- 11028862 TI - Bullous eruption associated with scabies: evidence for scabetic induction of true bullous pemphigoid. AB - Some patients with scabies develop bullae concomitantly with, or subsequently after, the occurrence of scabetic lesions. Although several immunofluorescence studies have demonstrated immunoglobulin deposition in the basement membrane zone of bullous lesions, it remained unclear whether these antibodies are directed to bullous pemphigoid antigens. We clearly show that two scabetic patients with bullous eruptions had circulating antibodies against BP180 and/or BP230 as determined by Western blotting analysis. This is the first report to demonstrate that at least some of the bullous eruptions occurring in scabetics are true bullous pemphigoid. PMID- 11028863 TI - Comparison of severity scoring of atopic dermatitis values and serum levels of eosinophil cationic protein and mast cell tryptase for routine evaluation of atopic dermatitis. AB - In this study the routine use of different parameters for evaluation of the overall therapeutic outcome in atopic dermatitis was investigated. The disease activity of 117 randomly selected hospitalized patients suffering from atopic dermatitis was routinely assessed using the Severity Scoring of Atopic Dermatitis (SCORAD) index on admission and at discharge. Serum levels of eosinophil cationic protein and mast cell tryptase were determined in parallel both on admission and at discharge. After a mean treatment period of 24+/-12 days a decrease in the SCORAD index from 47.6+/-19.5 to 7.7+/-8.2 was achieved (p<0.001). Serum levels of eosinophil cationic protein decreased from 22.8+/-19.7 microg/l to 15.4+/-17.5 microg/l, whereas serum tryptase levels did not change. However, there was no significant correlation between the changes in SCORAD, eosinophil cationic protein and tryptase in our cohort. Thus, routine determination of serum eosinophil cationic protein or tryptase levels, in addition to evaluation of disease activity using the SCORAD index, is not recommended in unselected patients with atopic dermatitis. PMID- 11028864 TI - Expression and functional role of co-stimulatory molecules in CD40+IL-4 stimulated B cells from atopic and non-atopic donors. AB - As immunological dysregulation is a possible key defect in atopic diseases, we have studied the expression and function of costimulatory molecules in atopic dermatitis (AD) patients compared with normal controls. Using flow cytometry, we showed that CD80 and CD86 are expressed at increased levels on human peripheral B cells in both groups after stimulation with anti-CD40 and interleukin 4 (IL-4), but to a significantly higher extent in the AD group. Furthermore, baseline expression of CD80 and CD86 on peripheral B cells was low in normal donors and increased in AD donors. To study the functional role of the costimulatory molecules in CD40+IL-4-stimulated peripheral mononuclear cells from normal and atopic donors, proliferation and IgE production were analysed in the presence of antibodies against the receptors of the costimulatory molecules. In the presence of either anti-CD28 or anti-CTLA-4, cell proliferation and IgE synthesis were significantly enhanced in the atopic group in anti-CD40+IL-4-stimulated peripheral mononuclear cells. These findings suggest that interaction of CD80 and CD86 with their receptors CD28 and CTLA-4 selectively promotes cell activation, including proliferation and IgE production in CD40+IL-4-stimulated peripheral blood mononuclear cells from atopic donors. It remains to be elucidated whether these changes are primary, based on the genetic background of atopics, or whether they are induced secondarily in the context of atopic pathology. PMID- 11028866 TI - Histological diagnosis of cutaneous "warty" carcinoma on a pre-existing HPV lesion. AB - A middle-aged man with an 8-year history of a fungating tumour mass on his thigh was histologically diagnosed as having an invasive "warty" carcinoma at the location of a pre-existing human papillomavirus (HPV) lesion. The tumour surface had a verruciform appearance with papillae containing fibrovascular cores. Many of the malignant cells displayed changes consistent with koilocytotic atypia. We noticed a greater degree of nuclear atypia in comparison with a verrucous carcinoma. Focally, some neoplastic cells demonstrated features of basaloid differentiation. Under in situ hybridization conditions, only HPV16 DNA, which is commonly associated with genital neoplasia, was detected selectively in rather superficial areas, corresponding to morphological evidence of HPV infection (i.e. koilocytotic atypia). It is worth noting that cutaneous location of such a warty carcinoma is very rare. PMID- 11028865 TI - Papular palmoplantar hyperkeratosis following chronic medical exposure to arsenic: human papillomavirus as a co-factor in the pathogenesis of arsenical keratosis? AB - This study presents the case of a 38-year-old patient from Pakistan with vitiligo, who developed multiple verrucous papules on the palms and soles several years after receiving "herbal treatment" from a travelling Indian doctor for a period of 12 months. Histopathological examination showed changes consistent with the diagnosis of arsenical keratosis. Molecularbiological examination of a skin biopsy detected an atypical human papillomavirus. This observation supports the concept of human papillomavirus as a co-factor in the pathogenesis of premalignant arsenic-induced skin tumours. PMID- 11028867 TI - Pigmented median raphe cysts of the penis. AB - Two cases of median raphe cysts of the penis with melanosis are presented. The presence of melanocytes was observed in the lining of the cysts by light and electron microscopy. The possible mechanism of the embryological development of the cysts is discussed in the context of the published literature. PMID- 11028869 TI - Porokeratosis in the elderly: a new subtype of disseminated superficial actinic porokeratosis. AB - In a review of all cases of porokeratosis histologically diagnosed in our Department during the period 1991-98 we found that 12 patients (22%) were in their seventh to ninth decade. In all 12 (2 males and 10 females) the age of onset of the disease varied between 58 and 89 years (mean age 68.6 years). The clinical picture was similar in all the patients, with the number of lesions varying from a few to 20-50 annular plaques 10-15 mm in diameter, localized mainly on the lower limbs. We suggest that our patients had a very mild form of disseminated superficial actinic porokeratosis confined to the extremities with an unusually late onset. This peculiar variety of late-onset disseminated superficial actinic porokeratosis may represent a type of immunosuppression induced porokeratosis where the pathologic clone for porokeratosis is present but remains latent until the amount of sun exposure, together with the physiological age-related lowering of immunocompetence, bring about its proliferation. PMID- 11028868 TI - Oral involvement in mycosis fungoides: report of two cases and a literature review. AB - Oral involvement is uncommon in cutaneous T-cell lymphomas and usually associated with poor prognosis. The clinicopathological and immunohistochemical findings of 2 new cases are described along with a literature review. The first patient had a 10-year history of mycosis fungoides when she developed lesions in the oral tissues. She died 6 months later despite treatment. The immunophenotype was CD3+, CD4+, CD8-, CD30-. The second patient had a mycosis fungoides for 5 years when she developed lesions in the uvula and oropharynx. She was treated with polychemotherapy and she is alive 5 years after oral involvement. The immunophenotype was CD3+, CD4-, CD8+, CD30-. There are conflicting reports about the prognosis in the CD8+ phenotype. The present cases and the literature review seem to indicate that in oral involvement the CD8- phenotype is not associated with a worse prognosis than the CD4+ subtype. However, it is necessary to study new cases to confirm this statement. PMID- 11028870 TI - Fulminant mucormycosis in a renal transplant recipient. PMID- 11028871 TI - Potentiating effect of imiquimod in the treatment of verrucae vulgares in immunocompromised patients. PMID- 11028872 TI - Electron beam therapy in patients with scleredema. PMID- 11028873 TI - Erythema multiforme-like subacute cutaneous lupus erythematosus: a new variety? PMID- 11028874 TI - Radiation myo-fasciitis. PMID- 11028875 TI - Superficial granulomatous pyoderma of the scrotum: an extremely rare cause of genital ulcer. PMID- 11028876 TI - Successful treatment of ulcerated necrobiosis lipoidica with mycophenolate mofetil. PMID- 11028877 TI - A suppurating fistula from a cement foreign body presenting as a tumour of the nail. PMID- 11028878 TI - Efficacy of cidofovir in the treatment of recalcitrant molluscum contagiosum in an AIDS patient. PMID- 11028879 TI - Fatal toxic epidermal necrolysis associated with ceftazidine and vancomycin therapy: a report of two cases. PMID- 11028880 TI - Randomized double-blind comparison of short-term itraconazole and terbinafine therapy for toenail onychomycosis. PMID- 11028881 TI - The Norwegian Arthroplasty Register: 11 years and 73,000 arthroplasties. AB - In 1985, the Norwegian Orthopaedic Association decided to establish a national hip register, and the Norwegian Arthroplasty Register was started in 1987. In January 1994, it was extended to include all artificial joints. The main purpose of the register is to detect inferior results of implants as early as possible. All hospitals participate, and the orthopedic surgeons are supposed to report all primary operations and all revisions. Using the patient's unique national social security number, the revision can be linked to the primary operation, and survival analyses of the implants are done. In general, the survival analyses are performed with the Kaplan-Meier method or using Cox multiple regression analysis with adjustment for possible confounding factors such as age, gender, and diagnosis. Survival probabilities can be calculated for each of the prosthetic components. The end-point in the analyses is revision surgery, and we can assess the rate of revision due to specific causes like aseptic loosening, infection, or dislocation. Not only survival, but also pain, function, and satisfaction have been registered for subgroups of patients. We receive reports about more than 95% of the prosthesis operations. The register has detected inferior implants 3 years after their introduction, and several uncemented prostheses were abandoned during the early 1990s due to our documentation of poor performance. Further, our results also contributed to withdrawal of the Boneloc cement. The register has published papers on economy, prophylactic use of antibiotics, patients' satisfaction and function, mortality, and results for different hospital categories. In the analyses presented here, we have compared the results of primary cemented and uncemented hip prostheses in patients less than 60 years of age, with 0-11 years' follow-up. The uncemented circumferentially porous- or hydroxyapatite (HA)-coated femoral stems had better survival rates than the cemented ones. In young patients, we found that cemented cups had better survival than uncemented porous-coated cups, mainly because of higher rates of revision from wear and osteolysis among the latter. The uncemented HA-coated cups with more than 6 years of follow-up had an increased revision rate, compared to cemented cups due to aseptic loosening as well as wear and osteolysis. We now present new findings about the six commonest cemented acetabular and femoral components. Generally, the results were good, with a prosthesis survival of 95% or better at 10 years, and the differences among the prosthesis brands were small. Since the practice of using undocumented implants has not changed, the register will continue to survey these implants. We plan to assess the mid- and long-term results of implants that have so far had good short-term results. PMID- 11028882 TI - Outcome after total hip arthroplasty: Part I. General health evaluation in relation to definition of failure in the Swedish National Total Hip Arthoplasty register. AB - The Swedish National Total Hip Arthroplasty register, which was started in 1979, is one of the oldest national quality registers in the world and consists of over 160,000 primary operations and 12,500 revisions. The register identifies risk factors for poor outcomes related to the patient, implant and surgical techniques. The end-point for failure, i.e., revision, does not provide information about the patient's general health after the primary procedure. The aim of this study was twofold. First, to validate the end-point for failure in the Swedish National Total Hip Arthroplasty register and secondly, to study general health after total hip arthroplasty. We validated the outcome of 1,056 primary THRs randomly selected from the Discharge register in Sweden by comparing the data to the Swedish THA register. These patients had answered the SF-36 and Nottingham Health Profile questionnaires. By comparing the clinical outcome, measured as general health, with the results obtained from the register, we evaluated the importance of the end-point for failure. We found that the end point was useful, but further evaluations are desirable. Patients operated on with hip replacement do very well up to 10 years postoperatively and those who are not revised have good general health. The findings in this study can be used as a reference for others as it shows results from a national register, with a random selection of the study cohort. PMID- 11028883 TI - Radiostereometric analysis in hip revision surgery--optimal time for index examination: 6 patients revised with impacted allografts and cement followed weekly for 6 weeks. AB - We revised the hip (6 stems and 5 sockets) with impacted morselized allografts and cement in 6 patients. We followed prosthetic migration by roentgen radiostereometric analysis (RSA) every 7th day for 6 weeks after the first (index) examination performed on the first postoperative day before mobilization. Most of the migration occurred during the first 2 weeks. In most cases more than half of the distal stem migration was seen between the 1st and 14th days. In all cases, the stem and socket migrations slowed down gradually and several prosthetic components had become stable after 5 weeks. We conclude that it is essential to perform the index RSA examination on the 1st or 2nd day after surgery and to state when and how weight bearing should be permitted. Otherwise it will be difficult to compare prosthetic migration in various studies and define normative values for migration predicting survival. PMID- 11028884 TI - Wear of ultra-high molecular weight polyethylene and polytetrafluoroethylene in a hip simulator: a dose-response study of protein concentration. AB - Charnley's laboratory wear studies of non-gamma sterilized polytetrafluoroethylene (PTFE) and polyethylene (PE) found that the PTFE to PE wear-rate ratio of 250:1 was much higher than the in-vivo wear ratio of 20:1. Tests of PTFE and PE in our laboratory showed a wear ratio of 150:1, using bovine serum as the lubricant and 190:1 with water as the lubricant. Our hypothesis was that the wear-rates of PTFE and PE cup materials were related to the concentration of protein in the serum. We studied the wear behavior of PTFE and PE cups in varied protein concentrations, using 4 femoral head sizes to validate the clinical range reported by Charnley. The PTFE wear-rates increased with increasing protein concentration and conversely, PE wear-rates decreased with increasing protein concentration. This inverse relationship made it possible to bring the wear ratio closer to the desired clinical wear ratio. We found that the clinically relevant PTFE/PE wear ratio corresponded to 3-10 mg/mL of protein in bovine serum. PMID- 11028885 TI - Slipped capital femoral epiphysis: early mechanical damage to the acetabular cartilage by a prominent femoral metaphysis. AB - On the basis of intraoperative observations in 13 consecutive adolescents (14 hips) with slipped capital femoral epiphysis (SCFE), we found that when the anterior femoral metaphysis was level with or extended past the epiphysis, it caused labrum and cartilage damage. As a result of an impingement between the metaphysis and the superomedial acetabular rim, the labrum revealed erosions, scars or tears. Further jamming of the metaphysis into the joint damaged the adjacent acetabular cartilage, varying from a partial- to a full-thickness cartilage loss. In all patients, the femoral head cartilage was intact; no avascular necrosis was present. Our findings suggest that arthrosis in SCFE can be triggered by early mechanical damage of the acetabular cartilage. PMID- 11028886 TI - Past incidence and future demand for knee arthroplasty in Sweden: a report from the Swedish Knee Arthroplasty Register regarding the effect of past and future population changes on the number of arthroplasties performed. AB - By combining data from the Swedish Knee Arthroplasty Register and Swedish census registers we have calculated the past age-specific incidence of primary knee arthroplasties and predicted the demand. During the last 20 years, osteoarthrosis has accounted for the largest increase in number of knee arthroplasties while operations for rheumatoid arthritis remained constant. The mean yearly number of operations between the periods 1976-1980 and 1996-1997 increased more than five fold, while only 6% of that increase could be explained by changes in the age profile of the population. Most operations were performed on persons of 65 years and older who also had the largest increase in incidence. By using the incidences for 1996 and 1997 and taking into account the expected future changes in the age profile of the Swedish population, we estimate that, in the absence of an effective preventive treatment, the number of knee arthroplasties will increase by at least one third until 2030. PMID- 11028887 TI - Tripled semitendinosus anterior cruciate ligament reconstruction with Endobutton fixation: a 2-3-year follow-up study of 35 patients. AB - We evaluated the clinical outcome of tripled semitendinosus tendon ACL reconstruction with femoral Endobutton (Acufex, Smith&Nephew, Andover, MA) and tibial Suture Washer (Acufex, Smith&Nephew, Andover, MA) (n 29) or post screw fixation (n 6) in 35 patients on an average of 28 (20-37) months after surgery. On the basis of IKDC criteria, 22 patients showed a normal or nearly normal knee function and 25 patients had a KT 1000 maximal manual side-to-side difference of < or =5 mm at follow-up. Subjectively, 28 patients graded their knee function as normal or nearly normal. The average Lysholm score was 88 points, average OAK score 90 points and average modified HSS score 93 points. 19 patients reached their preinjury level of activity at follow-up. The postoperative Lachman test was < or =1+ in 24 patients and 24 patients also showed an absent pivot shift. Significant bone tunnel enlargement occurred in 26 patients on the femoral side and in 23 patients on the tibial side. We found no correlation between bone tunnel enlargement and clinical outcome. The clinical outcome of tripled semitendinosus tendon ACL reconstruction with Endobutton fixation on the femoral side was not entirely satisfactory. The procedure was associated with a high incidence of bone tunnel enlargement in this series. PMID- 11028888 TI - Interaction between the ACL graft and MCL in a combined ACL+MCL knee injury using a goat model. AB - The optimal treatment for the MCL in the combined ACL and MCL-injured knee is still controversial. Therefore, we designed this study to examine the mechanical interaction between the ACL graft and the MCL in a goat model using a robotic/universal force-moment sensor testing system. The kinematics of intact, ACL-deficient, ACL-reconstructed, and ACL-reconstructed/ MCL-deficient knees, as well as the in situ forces in the ACL, ACL graft, and MCL were determined in response to two external loading conditions: 1) anterior tibial load of 67 N and 2) valgus moment of 5 N-m. With an anterior tibial load, anterior tibial translation in the ACL-deficient knee significantly increased from 2.0 and 2.2 mm to 15.7 and 18.1 mm at 30 degrees and 60 degrees of knee flexion, respectively. The in situ forces in the MCL also increased from 8 to 27 N at 60 degrees of knee flexion. ACL reconstruction reduced the anterior tibial translation to within 2 mm of the intact knee and significantly reduced the in situ force in the MCL to 17 N. However, in response to a valgus moment, the in situ forces in the ACL graft increased significantly by 34 N after transecting the MCL. These findings show that ACL deficiency can increase the in situ forces in the MCL while ACL reconstruction can reduce the in situ forces in the MCL in response to an anterior tibial load. On the other hand, the ACL graft is subjected to significantly higher in situ forces with MCL deficiency during an applied valgus moment. Therefore, the ACL-reconstructed knee with a combined ACL and MCL injury should be protected from high valgus moments during early healing to avoid excessive loading on the graft. PMID- 11028890 TI - Distal biceps tendon rupture operated on with the Boyd-Anderson technique: follow up of 9 patients with isokinetic examination after 1 year. AB - We operated on 9 patients for distal biceps tendon rupture using the Boyd Anderson technique. All patients were re-examined after at least 1 year using a questionnaire, radiographs, motion measurements and isokinetic testing. 2 patients had temporary radial nerve dysfunction and 7 patients had diminished forearm rotation. Elbow flexion strength was reduced by median 13% and supination strength by 19%. We think the Boyd-Anderson technique can be recommended, but slightly reduced strength and forearm rotation must be expected. PMID- 11028891 TI - Methods for determining the accuracy of radiostereometric analysis (RSA). AB - The problem of determining the noise level in any measuring system remains urgent. Radiostereometric analysis (RSA) is a radiographic system of unique accuracy which has applications in areas where minute motions or no motion at all occurs. Examples are micromotion between endoprostheses and bone and in fracture healing. We have determined the accuracy of the RSA system as applied to a clinical series of spinal fusions, where the conditions for RSA were not optimal. Using the usual test-retest methodology on a phantom, we showed that its accuracy can be grossly overestimated in the individual case. We found considerable variations in the accuracy in the individual case, depending on the rigid-body configuration. The overall accuracy, expressed as 3-D "vectors" for rotation and translation, respectively, correlated with the condition number, a method for characterizing the marker configuration. Indeed, the condition number explained as much as 92% of the variation in overall rotation. This condition number, however, cannot be used to analyze the accuracy of one degree of freedom of rotation alone. Mathematical simulation of the accuracy in the individual case of the individual dimension, using in-house software, showed that the accuracy (95% confidence) varied between 0.4 and 4.6 degrees of rotation about the transverse axis, corresponding to a clinical stress series of extension and flexion. PMID- 11028889 TI - Vastus lateralis fibrosis in habitual patella dislocation: an MRI study in 28 patients. AB - We studied 28 patients with habitual or recurrent dislocation of the patella with MRI of both thighs. Apart from the 2 patients whose dislocation could be related to trauma, we found signs of fibrosis of the vastus lateralis muscle in all the affected limbs of the 26 patients with an insidious onset of dislocation. This was seen as low signal intensity cords in the muscles in the T2 weighted image. Muscle degeneration was seen as high intensity signals in the T1 weighted image. In patients with unilateral disease, the vastus lateralis muscle of the affected side was hypotrophic, compared to that of the normal side. 2 patients underwent a biopsy of the affected muscle area. Histopathological examination revealed inflammatory cell infiltration, fibrosis, and muscle fiber degeneration. Fibrosis of the vastus lateralis muscle appears to be common in patients with habitual patella dislocation in our population, and may be the cause of the dislocation. Since release of such a contracture may be of value, MRI study of the thigh muscles is helpful in the evaluation of patients with this disorder. PMID- 11028892 TI - Biochemical bone markers and bone density in hip fracture patients: weak correlation in 106 women. AB - Biochemical markers of bone formation and bone resorption were measured preoperatively, within 18 hours after a hip fracture (n 106) and bone mineral density (BMD) was measured with quantitative computer tomography (QCT), dual energy x-ray absorptiometry (DXA) and heel ultrasound in 63 hip fracture patients. Patients with pertrochanteric fractures had more osteoporosis in all measurements than patients with femoral neck fractures. We found no differences in biochemical markers of bone formation or bone resorption in patients with femoral neck fractures and in those with pertrochanteric fractures. Correlations between biochemical bone markers and bone density, bone mass and stiffness on admission were weak. PMID- 11028893 TI - Role of multinuclear cells in granulation tissue in osteomyelitis: immunohistochemistry in 66 patients. AB - We investigated the origin of multinuclear cells (MNCs) in the granulation tissue in osteomyelitis by immunohistochemical techniques in 66 patients. 12 samples were analyzed for the presence of CD68, cathepsin K, CD11b and tartrate-resistant acid phosphatase (TRAP) activity. Many MNCs were present in the granulation tissue adjacent to a sequestrum. MNCs in contact with the sequestrum were also noted, however, no osteoblasts were found. Immunohistochemically, CD68, cathepsin K and TRAP were strongly expressed in most of the MNCs, while CD11b positive cells were not found. MNCs remote from and in contact with the sequestrum showed the same immunohistochemical features which are characteristic of osteoclasts. Further, MNCs in contact with the sequestrum had originally developed in the granulation tissue and directly infiltrated towards the sequestrum without cell to-cell interaction with osteoblasts. PMID- 11028894 TI - Proximal tibia--extensor mechanism composite allograft for revision TKA with chronic patellar tendon rupture. AB - Rupture of the patellar tendon following TKA is fortunately an uncommon complication with an incidence of 0.2-4% (Abril et al. 1995). Numerous options have been reported for treating this complication, including primary suture into a bone trough (Abril et al. 1995), autogenous tendon transfer (Cadambi and Engh 1992), and use of an artificial ligament (Fujikawa et al. 1994). Allograft patellar tendon has also been utilized either retaining the host patella and using a bone-tendon-bone graft or excising the host patella and using a quadriceps tendon-patella-patellar tendon-tibial tubercle graft (Emerson et al. 1990, Emerson et al. 1994, Zanotti et al. 1995, Booth et al. 1999). All of these techniques require structural integrity of the proximal tibia in the area of the tibial tubercle in order to attach the repair or graft to restore continuity of the extensor mechanism. When there is massive osteolysis and bone loss involving the proximal tibia, the situation becomes more complex and the previously described techniques are not advisable. In such cases, function can be obtained with a composite allograft of proximal tibia-patellar tendon-patella-quadriceps tendon. We describe a technique for dealing with this difficult clinical situation. PMID- 11028895 TI - Recurrent anterior dislocation of the hip joint--a case report. PMID- 11028896 TI - Pigmented villonodular synovitis of the wrist invading bone--a report of 2 cases. PMID- 11028897 TI - Unusual course of the extensor pollicis longus tendon associated with tenosynovitis, presenting as de Quervain disease--a case report. PMID- 11028898 TI - Is the paternal mononuclear cells' immunization a successful treatment for recurrent spontaneous abortion? AB - PROBLEM: Alloimmunization as a treatment for recurrent spontaneous abortion (RSA) is still controversial due to the lack of enough controls to evaluate its effectiveness. The present study was conducted to compare the live birth rate in the presence or absence of immunotherapy. METHOD OF STUDY: Ninety-two women with RSA (79 primary [PA] and 13 secondary aborters[SA]) received immunotherapy. Thirty-seven RSA couples not receiving paternal alloimmunization, constituted the "control" group. RESULTS: The pregnancy rate in alloimmunized was 58 vs 46% in the control group. The live birth increased from 71% in the controls to 88% after immunotherapy. The alloimmunization induced mixed lymphocyte reaction blocking factors (MLR BFs) in 79% of women. However, they were also present in 83% of immunized women experiencing a new abortion. CONCLUSION: These results indicate that alloimmunization may be useful in the treatment of RSA. PMID- 11028899 TI - Anti-DNA antibodies cross-reacting with laminin inhibit trophoblast attachment and migration: implications for recurrent pregnancy loss in SLE patients. AB - PROBLEM: Systemic lupus erythematosus (SLE), an autoimmune disease, is associated with reduced fetal survival, recurrent abortions, and other pregnancy complications. Some of the autoantibodies found in SLE bind to laminins (LNs), which play an important role in the implantation of the fertilized ovum in humans. METHOD OF STUDY: To elucidate the role of these specific autoantibodies, chorionic villous explants from 6 7-week-old human placentas were established as organ cultures on laminin-1 (LN-1), collagen IV (CN-IV) or uncoated culture dishes. The cultures were then exposed to a mouse monoclonal anti-DNA/anti-LN-1 antibody, to human polyclonal lupus antibodies cross-reacting with LN-1, a function-blocking polyclonal antibody to LN-1, polyclonal antibodies to CN-IV, or IgG control. RESULTS: The explants attached to LN-1 and CN-IV, but not to uncoated culture dishes. LN-1 promoted migration of trophoblast, whereas CN-IV promoted migration of fibroblast-like cells. Trophoblast attachment and migration were abolished in a dose-dependent manner by all three antibodies to LN-1, but not by antibodies to CN-IV or IgG control. Furthermore, the effect of anti-LN antibodies was abolished by preincubating them with LN-1. CONCLUSIONS: These studies suggest that anti-DNA antibodies cross-reacting with LNs may play a role in early pregnancy failure in SLE patients by interfering with placental implantation. PMID- 11028900 TI - Changes in cytokine production during and after normal pregnancy. AB - PROBLEM: The systemic T helper 1/T helper 2 (Th1/Th2) cytokine balance during normal human pregnancy is controversial, and observations about the balance in the postpartum period have only been reported for up to 3 months. METHOD: Whole blood, from 83 healthy pregnant women, 80 healthy postpartum women, and 31 healthy non-pregnant women was stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin, and the levels of cytokines in the supernatant were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The production of all measured cytokines decreased during pregnancy, especially in the second trimester. After delivery, interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) increased from 2 to 11 months postpartum, and IL-4 increased from 6 to 11 months postpartum. CONCLUSIONS: These data indicate that 1) decreases in production of both Th1-and Th2-type cytokines during pregnancy may be related to the pregnancy induced amelioration of autoimmune diseases: 2) increases in production of both Th1- and Th2-type cytokines in the postpartum period may be related to the postpartum aggravation of autoimmune diseases. PMID- 11028901 TI - Reconstitution of humoral immunity during pregnancy. AB - BACKGROUND: Recently, increasing attention has been paid to hormonal regulations of the immune system. MATERIALS AND METHODS: In this study, cord sera and the corresponding maternal sera were obtained at delivery. Sera from pregnant women were obtained at early, middle, and late stages of pregnancy. These sera were tested for titer and avidity of measles or mumps virus-specific immunoglobulin G (IgG) by means of a single-dilution, urea-denaturation enzyme-linked immunosorbent assay method. RESULTS: A positive and significant correlation was found between the titer and avidity of the virus-specific IgG, both in the cord sera and in the maternal sera. This correlation was established already at the early stage of pregnancy. There were no such correlations found in nonpregnant individuals. CONCLUSIONS: This is the first observation in human subjects that the avidity and concentration of the virus-specific IgG had a positive and significant correlation. Pregnancy must have some significant effects on the regulation of humoral immunity. PMID- 11028902 TI - TNFalpha expression in hyperplastic endometrium. AB - PROBLEM: Tumor necrosis factor-alpha (TNFalpha) is produced by the endometrium, and it has been shown that this cytokine has cyclic variations through the menstrual cycle. METHOD OF STUDY: In this study we assessed TNFalpha, estradiol and progesterone levels in the endometrium and serum of 21 patients with recent abnormal uterine bleeding (AUB). Eleven women showed histological diagnosis of endometrial hyperplasia, and ten women showed normal endometrium. RESULTS: The TNFalpha value in the serum of patients with hyperplasia and in normal endometrium did not show significant differences (64.8 +/- 21.4 vs 77.6 +/- 14.8 pg). The amounts of TNFalpha released by hyperplastic endometrial tissue were significantly higher than in control patients (258.8 +/- 78.0 vs 140.6 +/- 79.8 pg/g; P < 0.001). Immunohistochemistry of hyperplastic endometria showed a stronger staining than normal ones. Serum E2 levels in patients with hyperplastic endometrium were 49.5 +/- 31.6 ng/mL, and in controls were 107.3 +/- 66.0 ng/mL. The levels of estradiol released by hyperplastic endometria were significantly lower than in normal endometria (152.7 +/- 60.0 vs 279.9 +/- 178.7 ng/g; P < 0.003). CONCLUSIONS: Hyperplastic endometria produce higher amounts of TNFalpha with respect to controls. These findings may explain the frequent bleeding observed in the patients with endometrial hyperplasia, since TNFalpha promotes apoptosis and endometrial shedding. PMID- 11028903 TI - Regulation of immune functions by sperm-specific LDH and its differences with somatic isozyme in primary and secondary lymphocyte cultures. AB - PROBLEM: Sperm-specific lactate dehydrogenase-C4 (LDH-C4) is an autoantigen that produces experimentally induced autoimmune orchitis in testes. In the present study, immunological functions of B and T cells have been examined and compared after immunization with sperm-specific LDH and the LDH from somatic cells. METHODS: Three sets of experiments were performed. In the first set, effects of Balb/C LDH isozymes at 10(-3) - 1 L microg/well were investigated: (i) by mixed lymphocyte cultures (MLC) using C-57 B1/6 female cells as responders and AKR lymphocytes (irradiated) as stimulators, (ii) for regulatory T cell activity in MLC co-cultured along with Con-A-induced AKR lymphoblasts and (iii) for modulation of lymphocyte activation by PHA in vitro. In the second set of experiments, female mice (C-57 B1/6) were distributed in six groups for various treatments: i) saline (as vehicle), ii) adjuvant, iii) LDH-B4 (20 x 3 microg), iv) LDH-B4 (40 x 3 microg), v) LDH-C4 (20 x 3 microg), and v) LDH-C4 (40 x 3 microg). Mice were hyperimmunized with -B4 or -C4 (Balb/c) with a primary dose of 20 or 40 microg of protein per mouse, emulsified in Freund's complete adjuvant (FCA) and two identical doses in Freund's incomplete adjuvant (s.c.) within 22 days. Saline (group i) or adjuvant treated dams (group ii) served as controls. One week after the second booster, sera were tested for IgG response and lymphocytes harvested for polyclonal activation in vitro using LPS and Con-A as mitogens. In the third set of experiments, female Balb/c mice were divided into six groups as in the second experiment and immunized with a single primary dose of isogenic LDH-B4 or LDH-C4 at 20 or 40 microg of protein in FCA. On day 5, after sensitization with LDH, lymphocytes were evaluated for mitogenesis and for IgM production in vitro using LPS and Con-A as mitogens. RESULTS: i) Primary MLC(s) were non-specifically suppressed in the presence of 10(-3)- 1 L x microg allogenic LDH-C4 or -B4, although LDH-C4 tended to abolish MLC completely. But MLC co-cultured with blast cells was suppressed by LDH-C4 alone, indicating that sperm LDH suppresses induced formation of regulatory T cells. ii) FCA primed lymphocytes in situ were significantly inhibited for Con-A stimulation in vitro. Since LPS stimulation remained unaffected, it appeared that FCA is immunosuppressive for T cell proliferation alone. iii) Cells primed with LDH increased mitogenic activity of LPS several fold, although LDH-C4 was less effective than LDH-B4 in sensitization of B lymphocytes. iv) However, effect of Con-A in mitogenesis was dose-dependent, viz. cells primed at 20 x 3 microg of each isozyme overcame the immunosuppressive nature of FCA by bringing back the SI ( x 25) equivalent to saline primed cells, while pre-treatment of cells with 40 x 3 microg LDH-C4 abolished SI completely, indicating that -C4 primed cells were immunologically suppressed for Con-A stimulation. Such a response was markedly visible when allogenic LDH-C4 was used for hyperimmunization; lymphocytes challenged with somatic LDH under similar conditions did not react. Loss of T cell functions by LDH-C4 was confirmed in the presence of PHA in primary cultures. v) For antibody responses, although sperm LDH was highly reactive and dose-dependent, somatic LDH was also immunogenic for IgG production in serum to a lesser degree. Besides, IgM antibody was also discernible by two isozymes in LPS induced cultures. Significantly, -C4 primed cells at the higher dose, in comparison with the lower dose, were less responsive for IgM production. CONCLUSIONS: It is concluded that LDH(s) from sperm and somatic cells share functionally related antigenic epitopes that can generate/modify immune responses in vivo and in vitro with qualitative differences. However, immunosuppressive determinant of LDH-C4 is cell specific and dose selective. PMID- 11028904 TI - Immunoidentification of gonadotropin releasing hormone receptor in human sperm, pituitary and cancer cells. AB - PROBLEM: To generate monoclonal antibodies specific to the receptor of gonadotropin releasing hormone (GnRH) for immunoidentification of the GnRH receptor (RGnRH) in human sperm, pituitary and cancer cells. METHODS: Four oligopeptides corresponding to RGnRH amino acid residues 1-29, 182-193, 195-206 and 293-306 in the extracellular domains were synthesized, conjugated to hemocyanin from Keyhole Limpet and used as immunogens to generate specific monoclonal antibodies. Enzyme-linked immunosorbent assay was used for initial screening of hybridomas. RESULTS: A total of 15 hybrid cell lines secreting RGnRH specific monoclonal antibodies were initially established. By using some of these monoclonal antibodies as immunohistochemical probes, RGnRH was localized in the acrosomal region of human sperm, in the anterior pituitary tissue and in cancer cells of human ovarian and cervical origins. RGnRH was shown to have a molecular size of 45,000 +/- 5,000 kDa by Western blot assay. Expression of RGnRH mRNA in several human tissues and cancer cells was established by the reverse transcriptase-polymerase chain reaction method. CONCLUSION: RGnRH-specific monoclonal antibodies may be a valuable tool of identifying the presence of RGnRH in various normal and malignant human tissues. PMID- 11028905 TI - Corticosterone treatment of pregnant low dose endotoxin-treated rats: inhibition of the inflammatory response. AB - PROBLEM: Can the endotoxin-induced inflammatory response, underlying experimental pre-eclampsia, in pregnant rats be inhibited by corticosterone? METHOD OF STUDY: On day 10 of pregnancy, rats were implanted with pellets containing 25% corticosterone and 75% cholesterol (n = 10) or with 100% cholesterol-pellets (n = 10). On day 14 of pregnancy, rats were infused with either endotoxin (1.0 microg/kg bw) or saline. Three days later, they were sacrificed. Cryostat kidney sections were immunohistologically stained for the presence of neutrophils (PMN) and monocytes (MO) and the expression of inflammation-associated adhesion molecules. RESULTS: In cholesterol-treated rats, endotoxin significantly increased glomerular numbers of PMN and MO, glomerular expression of ICAM-1 and VCAM-1 and glomerular numbers of LFA-1 and VLA-4-positive cells as compared with saline. Corticosterone treatment significantly inhibited glomerular infiltration of PMN, MO and LFA-1 positive cells after endotoxin infusion. It did not affect glomerular ICAM-1 or VCAM-1 expression or numbers of VLA-4 positive cells. CONCLUSIONS: It is concluded that pre-treatment with corticosterone inhibits the low dose endotoxin-induced glomerular inflammatory reaction in pregnant rats, most likely by inhibiting LFA-1 expression, thereby decreasing the adhesiveness of inflammatory cells for activated endothelial cells. PMID- 11028906 TI - Immunocontraceptive efficacy of synthetic peptides corresponding to major antigenic determinants of chicken riboflavin carrier protein in the female rats. AB - PROBLEM: Earlier studies have demonstrated that antibodies directed towards the N terminal (residues 10-17) and C-terminal (residues 200-207) regions on chicken riboflavin carrier protein (RCP; 219 AA) are effective in pregnancy termination in rodents and sub-human primates. In the present study, the immunocontraceptive potential of three additional immunodominant sequences comprising of residues 33 49, 64 83 and 130-147 (CYA, CED and CGE peptides, respectively) of chicken RCP was investigated. METHOD OF STUDY: The three antigenic peptides were synthesized by using Fmoc chemistry. Oligoclonal antibodies were generated in rabbits. Bioneutralizing capacity of these peptides was assessed by passive and active immunoneutralization studies. RESULTS: All the three peptides-specific antisera recognized their cognate epitopes on native RCP. When the affinity purified peptide IgG were administered on three consecutive days to pregnant rats (on days 10, 11 and 12), it was observed that the rats injected with CED and CGE-IgG failed to deliver any pups whereas the animals which received CYA IgG delivered normal pups. Active immunization of fertile female rats with CED or CGE peptide conferred protection from pregnancy. CONCLUSIONS: These results demonstrate the presence of two additional stretches in chicken RCP which can serve as mini vaccines. PMID- 11028907 TI - Introduction: the regulation of eukaryotic transcription factor function. PMID- 11028908 TI - Mechanisms of action of transcription activation and repression domains. AB - Transcriptional regulators contain domains that either activate or repress transcription. Indeed, many cellular transcriptional regulators contain both activation and repression domains. Transcriptional regulators act at several stages in the transcription process, including assembly of the transcription complex, initiation and elongation. In order to influence these processes, the regulatory domains must interact with components of the transcription apparatus. This review will focus on our current understanding of the nature of transcriptional regulatory domains and their targets in the transcription machinery. PMID- 11028909 TI - Molecular basis of nutrient-controlled gene expression in Saccharomyces cerevisiae. AB - The ability of a unicellular organism to alter patterns of gene expression in response to nutrient availability is essential to its survival in a changing environment. How is the cell able to identify individual metabolites amongst a myriad of other similar molecules, and convert the information of its presence into a concerted change in the transcription of the genes required for the response to that metabolite? There is increasing evidence that the activity of transcription factors can be influenced directly by interaction with metabolites. A variety of mechanisms have been identified by which this type of gene regulation by small molecules can occur. PMID- 11028910 TI - Regulation of transcription factor function by phosphorylation. AB - Changes in protein phosphorylation represent a mechanism that is frequently employed by cells to regulate transcription factor activity. In response to alterations in the extracellular environment, signal transduction pathways target transcription factors, transcriptional coregulators and chromatin-modifying factors, leading to their phosphorylation by protein kinases or dephosphorylation by protein phosphatases. These modifications either positively or negatively regulate transcription factor activity to facilitate a program of gene expression that results in appropriate changes in cell behavior. Protein phosphorylation and dephosphorylation can directly regulate distinct aspects of transcription factor function, including cellular localization, protein stability, protein-protein interactions and DNA binding. The phosphorylation-dependent modulation of the activities of transcriptional coregulators and chromatin-modifying factors can also control transcription factor activity. Here we review recent studies that have led to a better understanding of the mechanisms by which protein phosphorylation and dephosphorylation governs transcription factor function. PMID- 11028911 TI - Regulation of gene expression by transcription factor acetylation. AB - In the nucleus, DNA is tightly packaged into higher-order structures, generating an environment that is highly repressive towards DNA processes such as gene transcription. Acetylation of lysine residues within proteins has recently emerged as a major mechanism used by the cell to overcome this repression. Acetylation of non-histone proteins, including transcription factors, as well as histones, appears to be involved in this process. Like phosphorylation, acetylation is a dynamic process that can regulate protein-DNA and protein protein interactions. Moreover, a conserved domain, the bromodomain, has been implicated in the binding of acetylated peptides, suggesting a role for acetylation in intracellular signalling. PMID- 11028912 TI - Nucleocytoplasmic shuttling of transcription factors. AB - To elicit the transcriptional response following intra- or extracellular stimuli, the signals need to be transmitted to their site of action within the nucleus. The nucleocytoplasmic shuttling of transcription factors is a mechanism mediating this process. The activation and inactivation of the transcriptional response is essential for cells to progress through the cell cycle in a normal manner. The involvement of cytoplasmic and nuclear accessory molecules, and the general nuclear membrane transport components, are essential for this process. Although nuclear import and export for different transcription factor families are regulated by similar mechanisms, there are several differences that allow for the specific activation of each transcription factor. This review discusses the general import and export pathways found to be common amongst many different transcription factors, and highlights a select group of transcription factors that demonstrate the diversity displayed in their mode of activation and inactivation. PMID- 11028913 TI - Regulation of transcription factors by protein degradation. AB - The level of transcription factors is tightly controlled by their rates of synthesis and degradation. Many critical factors are maintained at an appropriate level by targeted addition of ubiquitin and degradation via the proteasome. Whereas ubiquitination targets modified proteins for degradation, modification of substrates by the family of ubiquitin-like proteins does not target the proteins for degradation but can alter the stability and other properties of the modified proteins. Here we discuss the elaborate mechanisms that have evolved to allow specific recognition of substrates targeted for modification. Specific examples are discussed to illustrate the different mechanisms involved and the importance of regulated degradation in diseases such as cancer. PMID- 11028914 TI - Peripheral B cell survival. AB - Recent findings suggest that lymphocyte survival is a continuous active process and support the role of B cell receptor engagement in B cell survival. In this context the conflict of survival interests between the diverse B cells gives rise to a pattern of interactions which mimics the behavior of complex ecological systems. In response to competition lymphocytes modify their survival requirements and diverge to occupy different immunological niches through differentiation. Thus naive and memory-activated B cell populations show independent homeostatic regulation. We discuss how niche differentiation allows the coexistence of different cell types and guarantees both repertoire diversity and efficient immune responses. PMID- 11028915 TI - Molecular mechanisms involved in cisplatin cytotoxicity. AB - cis-diamminedichloroplatinum(II) or cisplatin is a DNA-damaging agent that is widely used in cancer chemotherapy. Cisplatin cross-links to DNA, forming intra- and interstrand adducts, which bend and unwind the duplex and attract high mobility-group domain and other proteins. Presumably due to a shielding effect caused by these proteins, the cisplatin-modified DNA is poorly repaired. The resulting DNA damage triggers cell-cycle arrest and apoptosis. Although it is still debatable whether the clinical success of cisplatin relies primarily on its ability to trigger apoptosis, at least two distinct pathways have been proposed to contribute to cisplatin-induced apoptosis in vitro. One involves the tumour suppressor protein p53, the other is mediated by the p53-related protein p73. Coupling cisplatin damage to apoptosis requires mismatch repair activity, and recent observations further suggest involvement of the homologous recombinatorial repair system. At present it is generally accepted that abortive attempts to repair the DNA lesions play a key role in the cytotoxicity of the drug, and loss of the mismatch repair activity is known to cause cisplatin resistance, a major problem in antineoplastic therapy. Clearly, a better understanding of the signalling networks involved in cisplatin toxicity should provide a rational basis for the development of new therapeutic strategies. PMID- 11028916 TI - New insights into copper monooxygenases and peptide amidation: structure, mechanism and function. AB - Many bioactive peptides must be amidated at their carboxy terminus to exhibit full activity. Surprisingly, the amides are not generated by a transamidation reaction. Instead, the hormones are synthesized from glycine-extended intermediates that are transformed into active amidated hormones by oxidative cleavage of the glycine N-C alpha bond. In higher organisms, this reaction is catalyzed by a single bifunctional enzyme, peptidylglycine alpha-amidating monooxygenase (PAM). The PAM gene encodes one polypeptide with two enzymes that catalyze the two sequential reactions required for amidation. Peptidylglycine alpha-hydroxylating monooxygenase (PHM; EC 1.14.17.3) catalyzes the stereospecific hydroxylation of the glycine alpha-carbon of all the peptidylglycine substrates. The second enzyme, peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL; EC 4.3.2.5), generates alpha-amidated peptide product and glyoxylate. PHM contains two redox-active copper atoms that, after reduction by ascorbate, catalyze the reduction of molecular oxygen for the hydroxylation of glycine-extended substrates. The structure of the catalytic core of rat PHM at atomic resolution provides a framework for understanding the broad substrate specificity of PHM, identifying residues critical for PHM activity, and proposing mechanisms for the chemical and electron-transfer steps in catalysis. Since PHM is homologous in sequence and mechanism to dopamine beta-monooxygenase (DBM; EC 1.14.17.1), the enzyme that converts dopamine to norepinephrine during catecholamine biosynthesis, these structural and mechanistic insights are extended to DBM. PMID- 11028917 TI - Penaeidins, a family of antimicrobial peptides from penaeid shrimp (Crustacea, Decapoda). AB - The production of antimicrobial peptides represents a first-line host defense mechanism of innate immunity that is widespread in nature. Only recently such effectors were isolated in crustacean species, whereas numerous antimicrobial peptides have been characterized from other arthropods, both insects and chelicerates. This review presents findings on a family of antimicrobial peptides, named penaeidins, isolated from the shrimp Penaeus vannamei. Their structure and antimicrobial properties as well as their immune function will be discussed through analyses of penaeidin gene expression and peptide distribution upon microbial challenge. PMID- 11028918 TI - Signaling pathways between the plasma membrane and endoplasmic reticulum calcium stores. AB - This review discusses multiple ways in which the endoplasmic reticulum participates in and is influenced by signal transduction pathways. The endoplasmic reticulum provides a Ca2+ store that can be mobilized either by calcium-induced calcium release or by the diffusible messenger inositol 1,4,5 trisphosphate. Depletion of endoplasmic reticulum Ca2+ stores provides a signal that activates surface membrane Ca2+ channels, a process known as capacitative calcium entry. Depletion of endoplasmic reticulum stores can also signal long term cellular responses such as gene expression and programmed cell death or apoptosis. In addition to serving as a source of cellular signals, the endoplasmic reticulum is also functionally and structurally modified by the Ca2+ and protein kinase C pathways. Elevated cytoplasmic Ca2+ causes a rearrangement and fragmentation of endoplasmic reticulum membranes. Protein kinase C activation reduces the storage capacity of the endoplasmic reticulum Ca2+ pool. In some cell types, protein kinase C inhibits capacitative calcium entry. Protein kinase C activation also protects the endoplasmic reticulum from the structural effects of high cytoplasmic Ca2+. The emerging view is one of a complex network of pathways through which the endoplasmic reticulum and the Ca2+ and protein kinase C signaling pathways interact at various levels regulating cellular structure and function. PMID- 11028919 TI - Signaling by reactive oxygen species in the nervous system. AB - Free radicals and reactive oxygen species (ROS) are involved in a variety of different cellular processes ranging from apoptosis and necrosis to cell proliferation and carcinogenesis. Cells contain multiple sites for ROS production and a few mechanisms for their degradation. Which of these sites is activated by a given stimulus may play a role in dictating the subsequent downstream effects of the ROS generated on cellular function. Even when the ultimate outcome is similar, such as when ROS production leads to cell death, the specific cellular changes can be quite different depending on the initial stimulus and the type of cell involved. These data, along with the evidence that ROS can modify a number of intracellular signaling pathways including protein phosphatases, protein kinases and transcription factors, suggest that the majority of the effects of ROS on cells are through their actions on signaling pathways rather than via nonspecific damage of intracellular macromolecules. PMID- 11028920 TI - Localization and analysis of nonpolar regions in onconase. AB - A detailed analysis of the composition and properties of hydrophobic nuclei and microclusters has been carried out for onconase. Two main hydrophobic nuclei in the onconase structure were detected. Their composition and shape were found to be very similar to those of RNase A, in accordance with the predictions made. The nuclei in onconase are more compact, the side-chain atoms of residues included in the nuclei in onconase form more contacts with the environment than in RNase A. The hydrophobic nuclei should be considered as individual structural units along with elements of the secondary structure. Differences in composition and conformation of exposed loops between onconase and RNase A were found. The additional hydrophobic clusters attached to the nuclei in onconase might be involved in the fixation of an appropriate conformation of site(s) for manifestation of the biological activity of onconase. A comparison of amphibian representatives of the RNase A superfamily was also made. The results obtained suggest that the availability of nonpolar residues in established key positions of amino acid sequences determines the characteristic fold of homologous proteins and the structure of the active site cleft. PMID- 11028921 TI - Differential basal synthesis of Hsp70/Hsc70 contributes to interindividual variation in Hsp70/Hsc70 inducibility. AB - The source of intraspecies variation in the expression of heat shock proteins (HSPs) remains unresolved but could shed light on differential stress tolerance and disease susceptibility. This study investigated the influence of variable basal HSP synthesis on differential inducibility of HSP synthesis. Basal and heat induced synthesis of the major HSP families in peripheral blood monocytes from healthy donors (n = 42) were analysed using biometabolic labelling and densitometry. Basal Hsp70/Hsc70 synthesis and percentage induction of Hsp70/Hsc70 synthesis were significantly correlated (r = -0.57, p < 0.0001), and described most accurately by an exponential decay equation (R = 0.68, R2 = 0.46). This regression equation suggests that increasing levels of basal Hsp70/Hsc70 synthesis are accompanied by an exponential decrease in the percentage induction of Hsp70/Hsc70 synthesis. The model fits data from European and non-European population groups independently, although both coefficients in the regression equation were larger for non-Europeans. This implies population group as an additional factor influencing differential HSP expression. The differential inducibility of Hsp70/Hsc70 due to variable basal synthesis of Hsp70/Hsc70 and based upon population group may contribute to differential stress tolerance or disease susceptibility. PMID- 11028922 TI - Highly efficient cell-mediated gene transfer using non-viral vectors and FuGene6: in vitro and in vivo studies. AB - The present study was undertaken to develop an efficient non-viral gene delivery system for cardiovascular gene therapy. We investigated transfection efficiency and toxic properties of the new transfection reagent, FuGene6, and compared it with two other transfection reagents, Tfx-50 and LipoTaxi. For in vivo experiments, the plasmid was delivered intramuscularly via transplantation of fibroblasts transfected with plasmid and FuGene6. Conditions for efficient gene delivery were initially studied in vitro. Human and rabbit fibroblasts were isolated from skin, cultured and transfected with phVEGF165 or pCMVbeta gal plasmids, coding for vascular endothelial growth factor (VEGF) or beta galactosidase, respectively. The effect of the DNA amount and the DNA:transfection reagent ratio on plasmid uptake were studied. Of the transfection reagents tested, only FuGene6 provided high-efficiency and dose dependent plasmid transfer both for cell-localised (beta-galactosidase) and secreted (VEGF) gene products. When analysed with an MTT assay, FuGene6 showed no toxicity at low doses. Optimised conditions were applied for in vivo reporter gene delivery. Rabbits were injected intramuscularly with ex vivo-transfected fibroblasts. As in in vitro studies, ex vivo-transfected fibroblasts showed highly efficient gene expression in vivo. Tissue sections were analysed with macrophage-specific immunostaining. No signs of inflammation were seen in the region of fibroblast injection. This study demonstrates that FuGene6 is a highly efficient transfection reagent that may be useful for in vitro non-viral transfection of primary human and rabbit fibroblasts and for in vivo therapeutic non-viral gene delivery. PMID- 11028923 TI - Application of ICSI for cryptorchid therapy. AB - Recent advances are provided in assisted reproductive technology and application of ICSI as treatment for infertile patients with previous cryptorchidism. Fertility rates were 25-81% and 8-48% in patients with unilateral and bilateral cryptorchidism, respectively. With the advent of assisted reproductive technology, pregnancy was not possible without sperm. Research in humans has resulted in pregnancy and live births after the injection of haploid germ cells (round spermatids) into the oocytes. In the mouse tetraploid germ cells (primary spermatocytes), nuclei have been injected into oocytes, which undergo meiosis, forming an embryo and live young. In animal models, round spermatids from both cryptorchid and previously cryptorchid testes can initiate normal embryo development when injected into mature oocytes. These cells may be used as substitutes for mature sperm. However, cryptorchidism in humans is often associated with inherent defects of spermatogenesis. With successive breakthroughs in treatment of impaired fertility associated with cryptorchidism, there is an increased risk of genetic abnormalities in the progeny. It is incumbent on clinicians who are involved in reproductive medicine to proceed cautiously with the development and application of assisted reproductive techniques to avoid creating future generations of genetically abnormal individuals. PMID- 11028924 TI - Assessment of two devices for in vitro preparation of human sperm. AB - The aim of the study was to assess and compare the efficiency of the ZSC-II versus the Sperm Select techniques in preparing human sperm from normozoospermic patients for use in intrauterine insemination and other assisted reproductive technologies. Twenty-five patients were included in the study. Semen was collected at intercourse and processed via 2 sperm preparation methods. Recovery of motile sperm and other sperm qualitative measurements were evaluated before and after ZSC-II and Sperm Select sperm preparation. Sperm qualitative measurements were not significantly different after the ZSC-II and Sperm Select procedures (p > .05). Differences (p < .05) were noted between the 2 procedures in the number of sperm recovered. A higher survival rate (longevity test, 72 h) was observed after ZSC-II sperm preparation and recovery. The ZSC-II procedure yielded higher total motile sperm than the Sperm Select. The superiority in longevity may suggest possible advantages in obtaining higher fertilization and pregnancy rates. PMID- 11028925 TI - Laparoscopic varicocelectomy in adolescents using an ultrasonically activated scalpel. AB - Ten laparoscopic varicocelectomies were performed using an ultrasonically activated scalpel to divide spermatic vessels in adolescents. The mean operation time was 35 (25-46) min. There were no intraabdominal viscelar or vascular complications, and no postoperative analgesia was required. These findings suggest that laparoscopic varicocelectomy using an ultrasonically activated scalpel may be a valuable treatment in the adolescent. PMID- 11028926 TI - Surrogate pregnancies in patients with Mayer-Rokitansky-Kustner-Hauser syndrome and severe teratozoospermia. AB - The challenge of the case presented was threefold: congenital absent uterus (Mayer-Rokitansky-KustnerHauser syndrome), a request for surrogacy, and teratozoospermia in the husband (6% normal morphology according to the Tygerberg strict criteria). Surrogacy was dealt with by means of guidelines proposed by the SA Law Commission after a surrogate mother was found. The gestational carrier was synchronized with the genetic donor with congenital absent uterus, the main aim being assisted reproduction. Two gamete intrafallopian transfer (GIFT) procedures were performed, both unsuccessful. Poor fertilization of excess GIFT oocytes was also observed. Three intracytoplasmic sperm injection procedures were then performed. The first two were unsuccessful and the third ensued in a singleton pregnancy. Hormonal support (progesterone and estradiol valerate until 12 weeks pregnancy duration) was given. The pregnancy resulted in the normal delivery of a healthy male infant at full term. Psychological support was given to mother and surrogate throughout pregnancy, as well as thereafter. The success of this case gives hope to infertile couples with severe infertility factors. PMID- 11028927 TI - Does seminal fluid viscosity influence sperm chromatin integrity? AB - A retrospective study was undertaken to investigate whether viscosity alters sperm chromatin integrity. Semen samples were obtained from 269 men attending the infertility clinic. The viscosity was measured quantitatively by needle and syringe method and the viscosity ratio was calculated against distilled water. The chromatin integrity was evaluated by in vitro decondensation test using 1% SDS and 6 mM EDTA. According to the viscosity ratios the samples were divided into 2 groups: I, normal (ratio < 9, n = 239): and II, abnormal (ratio > 9, n = 30) viscosity. Chromatin integrity was significantly lower in the group with higher viscosity. Significant decrease in sperm count and motility were seen in group II as compared to group I. Thus, hyperviscosity of seminal fluid alters the sperm chromatin integrity. PMID- 11028928 TI - Galvanotactic response of mouse epididymal sperm: in vitro effects of zinc and diethyldithiocarbamate. AB - This study was conducted to evaluate the galvanotactic response of mouse epididymal sperm as well as the in vitro effects of Zn-acetate and diethyldithiocarbamate (DEDTC) on the galvanotaxis of sperm. The galvanotaxis of sperm was observed in a direct current (DC) electric field between 0 and 10 V/cm. The sperm were treated with Zn-acetate or DEDTC at concentrations of 0.02, 0.20, and 2.00 mM before the observations. The sperm exhibited galvanotaxis toward both cathode and anode in the fields between 1 and 9 V/cm. The number of sperm that accumulated at both electrodes was found to reach the highest level between 5 and 7 V/cm. In comparison with the results from untreated sperm, the number of accumulated sperm at the electrodes increased with the addition of 0.20 and 2.00 mM Zn-acetate, but decreased with the addition of DEDTC at all 3 concentrations. The galvanotaxis of mouse epididymal sperm is related to the field strengths, and the guidance of sperm migration may be influenced by the sex chromosomes. The galvanotaxis of the sperm may be improved by addition of Zn-acetate but depressed by DEDTC in the presence of DC electric fields. PMID- 11028929 TI - Poor quality of sperm as it affects repeated early pregnancy loss. AB - A study was carried out to determine whether males contribute to repeated early pregnancy loss. Semen samples were analyzed from proven-fertile men (n = 51 group I) and from men whose partners presented with early pregnancy loss (>3 first trimester abortions, n = 32 group II). Routine analysis, sperm function tests, and ultrastructural studies of sperms were carried out. Female factor could be identified in 25 (78%) couples, and in 7 (22%) no cause either male or female could be identified and the semen analysis was normal. Percent morphologically normal did not differ significantly between the groups, but increased sperm head abnormalities were seen. The functional tests were all normal except for a significant decrease in the capacity of nuclear chromatin to decondense in vitro. The ultrastructural studies showed defects of chromatin condensation and irregular nuclei with vacuoles. This study points to the loss of chromatin integrity as a possible contributing factor from males to early pregnancy loss. PMID- 11028930 TI - Ultrastructural interrelationship between the pineal gland and the testis in the male rat. AB - The ultrastructural interrelationship between the pineal gland and testis was evaluated in the rat. Wistar rats were divided into 6 groups. Groups I and II were sham-orchidectomized and orchidectomized rats, respectively. Rats in group III were orchidectomized and daily injected with testosterone propionate (TP) for 1 month. Groups IV and V were sham-pinealectomized and pinealectomized, respectively. Group VI was pinealectomized and daily injected with melatonin for 2 months. All animals were anesthetized with ketamine for fixation by vascular perfusion. Pineal glands of groups I, II, and III and the testes of groups IV, V, and VI were removed and weighed. All specimens were examined by electron microscopy. Orchidectomy caused an increase of lipid droplets, cytoplasmic dense bodies, and lysosomes. Rough endoplasmic reticulum, Golgi apparatus, and mitochondria were extensive in the cytoplasm. TP administration to orchidectomized rats resulted in formation of less extensive lipid droplets and mitochondria. In pinealectomized rats, golgi complex, mitochondria, and enlarged smooth endoplasmic reticulum were extensive in the cytoplasm of Leydig cells. Formation of cytoplasmic secretory granules and osmiophilic bodies was observed. Testicular weight increased compared to group IV. Melatonin decreased testicular weight in comparison to group V and prevented ultrastructural changes. Pinealectomy and orchidectomy caused hyperactivity in Leydig cells and pinealocytes, respectively, which suggests a mutual relationship between the pineal gland and testis in the rat. PMID- 11028931 TI - Stages in reproduction particularly vulnerable to xenobiotic hazards and nutritional deficits. AB - Biochemical research has identified many failures in reproductive processes with specific nutrient deficits, xenobiotics and some infectious illnesses. This has led to some effective safeguards. During meiosis and fertilization, as genetic material divides and rearranges, it is exposed and open to mutation. A nutritionally unfavourable environment is a major risk factor. At stages of rapid cell division, differentiation and organisation, as in the embryo and later in the fetal brain, the child's survival, completeness and future health and ability are at stake. From months before conception, reproduction needs preparing for, especially with today's environmental pollution, even entering the foodchain. Care from before conception can contribute not only to the child's healthy basis for life, full development of brain, eyesight and other complex attributes, but also to the health of at least the subsequent generation. Since the female baby's oocytes are being formed while she is still in the womb, the grandmother's nutritional status, around the time of conceiving a daughter, can permanently affect a grandchild. Recent insights into evolution, particularly of the brain, give us fresh indications of dietary needs to fulfil human potential for health and acuity. Despite the hazards, nature is remarkably successful. This paper is not designed to alarm but to help attainment of full genetic potential. With healthy parents serious malformations are a low percentage. The numbers of babies with avoidable disorders, however, calls urgently for action, especially in our own inner cities and in developing countries where there is inadequate nutrition. Action will more than justify itself, including financially. It will reward handsomely. PMID- 11028932 TI - Potential role of P-glycoprotein in affecting hepatic metabolism of drugs. PMID- 11028933 TI - Molecular cloning of a Na+-dependent nucleoside transporter from rabbit intestine. AB - PURPOSE: Substantial species differences in the transport kinetics of nucleosides and therapeutic nucleoside analogs have been observed in various experimental systems. To explain these differences at a molecular level, it is necessary to clone the relevant transporters and examine their functional characteristics in heterologous expression systems. The goal of the present study was to clone the nucleoside transporters present in rabbit, an important preclinical animal model, and to functionally characterize the clone(s). METHODS: A Polymerase Chain Reaction (PCR)-based homology cloning approach in conjunction with Rapid Amplification of cDNA Ends (RACE) was used to isolate a full-length cDNA. Characterization of this transporter was accomplished through heterologous expression in Xenopus laevis oocytes. RESULTS: A full-length cDNA encoding a purine-selective, Na+-dependent nucleoside transporter, rbSPNT1, was isolated from rabbit small intestine. The encoded protein is 658 amino acid residues in length. Hydropathy analysis suggests that rbSPNT1 has 11 to 14 transmembrane domains. In Xenopus laevis oocytes expressing rbSPNT1, the uptake of uridine and inosine was enhanced significantly; uridine transport was inhibited by purine, but not pyrimidine nucleosides. mRNA transcripts for rbSPNT1 were detected primarily in intestine, lung, and kidney and at lower levels in liver, brain, and heart. CONCLUSIONS: A full-length functional nucleoside transporter was cloned. Sequence analysis and functional characterization suggest that rbSPNT1 is the rabbit homolog of the purine-selective nucleoside transporter, N1. The cloned rbSPNT1 can be used to understand the molecular mechanisms responsible for the observed species differences in the transport of nucleosides and therapeutic nucleoside analogs. PMID- 11028934 TI - An economic cost analysis of oral ganciclovir prophylaxis for the prevention of CMV disease. AB - PURPOSE: The study conducted an economic cost analysis of oral ganciclovir prophylaxis in preventing cytomegalovirus (CMV) disease for AIDS patients in a randomized clinical trial setting. METHODS: Data were generated from patient interviews, medical records, and case reports from a multi-center, randomized, double-blind, and placebo-controlled pharmacoeconomic study appended to a clinical trial. The outcomes were measured in monthly cost per patient. Various cost functions were tested in the context of sample-selection model (SSM) and two part model (TPM), and were estimated using both the ordinary least squares (OLS) and the bounded influence estimation (BIE) methods. RESULTS: The use of informal caregiver services did not differ significantly between patients in the treatment group and those in the placebo group. The OLS estimates for the ganciclovir prophylaxis arm showed a reduced, but statistically insignificant use of formal care in both outpatient and inpatient settings. The BIE results for the ganciclovir prophylaxis arm, in contrast, showed a significant reduction of 27% in hospital cost among hospital users, and 44% among the total sample of AIDS patients. The monthly total cost function also identified a decreasing but insignificant trend due to the treatment effect. CONCLUSIONS: At the methodological level, this study demonstrated the value of employing more rigorous econometric techniques in identifying subtle treatment effects on cost outcomes from clinical trial data in the economic assessment of medical technologies. At the empirical level, the study concluded that beyond its demonstrated efficacy of preventing CVM disease among AIDS patients, ganciclovir prophylaxis did not lead to additional health care costs, other than the cost of the drug therapy. PMID- 11028935 TI - Quantitative correlation between initial dissolution rate and heat of solution of drug. AB - PURPOSE: The aim of this study is to estimate the initial dissolution rate of drug substances by isothermal microcalorimetry. A theory was presented on the basis of Gibbs free energy and the Noyes-Whitney equation. METHODS: Polymorphic forms and quenched glass of indomethacin, and some different crystallinity samples of terfenadine were used. The heats of solution of samples were measured by isothermal microcalorimetry. The initial dissolution rates of samples were measured by rotating disk method at 25 degrees C. RESULTS: Each drug showed a linear correlation between the heats of solution and the logarithms of initial dissolution rate, irrespective of their different crystal structure, such as polymorphic forms and crystallinity. The logarithms of initial dissolution rates were well correlated with the degree of crystallinity obtained by the isothermal microcalorimetry. CONCLUSIONS: The initial dissolution rates of drug substances could be estimated quantitatively from the heats of solution as estimated from the present theory. Isothermal microcalorimetry was extremely useful for the estimation of the initial dissolution rates of polymorphs and of partially crystalline samples. PMID- 11028936 TI - Temperature dependence of bimolecular reactions associated with molecular mobility in lyophilized formulations. AB - PURPOSE: We studied the temperature dependence of acetyl transfer between aspirin and sulfadiazine, a bimolecular reaction, in lyophilized formulations at temperatures near the glass transition temperature (Tg) and NMR relaxation-based critical mobility temperature (Tmc), to further understand the effect of molecular mobility on chemical degradation rates in solid pharmaceutical formulations. The temperature dependence of the hydrolysis rates of aspirin and cephalothin in lyophilized formulations was also studied as a model of bimolecular reactions in which water is a reactant. METHODS: Degradation of lyophilized aspirin-sulfadiazine formulations containing dextran and various amounts of water at temperatures ranging from 1 degrees C to 80 degrees C was analyzed by HPLC. The degradation of cephalothin in lyophilized formulations containing dextran and methylcellulose was also analyzed at temperatures ranging from 10 degrees C to 70 degrees C. RESULTS: Acetyl transfer in lyophilized aspirin--sulfadiazine formulations containing dextran exhibited a temperature dependence with a distinct break around Tmc, which may be ascribed to a change in the translational mobility of aspirin and sulfadiazine molecules. The hydrolysis of aspirin and cephalothin in lyophilized formulations, which is also a bimolecular reaction, did not show a distinct break, suggesting that water diffusion is not rate-limiting. CONCLUSIONS: The diffusion barrier of water molecules in lyophilized formulations appears to be smaller than the activational barrier of the hydrolysis of aspirin and cephalothin based on the results of this study that the temperature dependence of the hydrolysis rate is almost linear regardless of Tmc and Tg. On the other hand, the diffusion barrier of aspirin and sulfadiazine molecules appears to be comparable to the activational barrier of the acetyl transfer reaction between these compounds, resulting in nonlinear temperature dependence. PMID- 11028937 TI - Effects of lyophilization on the physical characteristics and chemical stability of amorphous quinapril hydrochloride. AB - PURPOSE: To prepare amorphous quinapril hydrochloride (QHCl) by lyophilization and to compare its physical characteristics and chemical stability as a function of the initial pH of the pre-lyophilized solution. METHODS: Amorphous QHCl samples were prepared by lyophilization from aqueous solutions. Solid-state characteristics were evaluated by DSC, PXRD, and optical microscopy. Chemical degradation was monitored by an HPLC assay. RESULTS: Amorphous QHCl samples obtained from lyophilization exhibited variable glass transition temperatures, depending on the pH and/or concentration of the starting aqueous solutions. Neutralized quinapril (Q) in the amorphous form, which has a Tg of 51 degrees C, lower than that of its HCl salt (91 degrees C), was significantly more reactive than QHCl at the same temperature. The Tg of lyophilized samples prepared at various initial pH values correlated well with values predicted for mixtures of QHCl and Q. Their different reaction rates were related to their glass transition temperature, consistent with the results from earlier studies obtained with amorphous samples made by precipitation from an organic solution and grinding of the crystal solvate. CONCLUSIONS: Lyophilization of different QHCl solutions produces mixtures of amorphous QHCl and its neutralized form Q, with Tg values intermediate to the values of QHCl and Q. As the fraction of Q increases the overall rate of chemical degradation increases relative to QHCl alone, primarily due to the increase in molecular mobility induced by the plasticizing effects of Q. PMID- 11028938 TI - Thermodynamics of binding of neutral molecules to sulfobutyl ether beta cyclodextrins (SBE-beta-CDs): the effect of total degree of substitution. AB - PURPOSE: To understand the role of degree of substitution on binding of molecules to beta-Cyclodextrins (beta-CDs) with varying degrees of sulfobutyl ether (SBE) substitution. METHODS: Using UV spectroscopy, complexation constants of molecules to SBE-beta-CDs were estimated as a function of temperature, allowing for calculation of thermodynamic parameters, including the enthalpy and entropy of binding. RESULTS: Binding constants of various molecules to SBE-beta-CDs did not show a uniform trend to total degree of SBE substitution. However, a distinct pattern was observed with the enthalpy and entropy of complexation. The results showed the complexation of substrates to SBE-beta-CDs to be more entropy-favored as the number of SBE groups increased. This favorable entropy of interaction was compensated by a less favorable enthalpy of interaction. CONCLUSIONS: Enthalpy and entropy of complexation provided additional insight into the role that the alkylsulfonate groups may play in the complexation of molecules with SBE-beta CDs. PMID- 11028939 TI - Deformation behaviors of tolbutamide, hydroxypropyl-beta-cyclodextrin, and their dispersions. AB - PURPOSE: The deformation behaviors of compressed freeze-dried and spray-dried tolbutamide/hydroxypropyl-beta-cyclodextrin molecular dispersions were evaluated and compared with similarly prepared tolbutamides (TBM), hydroxypropyl-beta cyclodextrins (HP-beta-CD) and as their physical dispersions. METHODS: TBM, HP beta-CD, and their 1:1 molecular dispersions were prepared by freeze-drying and spray-drying, and physical dispersions of TBM and HP-beta-CD were blended. Deformation properties of the prepared materials were evaluated by using a compaction simulator and constants derived from Heckel plots. Molecular dynamics (MD) simulations were performed in order to gain a molecular-level view on the deformation behavior of TBM-HP-beta-CD inclusion complex. RESULTS: The freeze dried TBM polymorphic form II was less prone to overall particle deformation than the spray-dried stable form I. Formation of molecular dispersions decreased the plastic and elastic behaviors of these materials. Also, the MD simulations showed a reduced molecular flexibility of the TBM-HP-beta-CD inclusion complex, as compared to HP-beta-CD. CONCLUSIONS: The formation of TBM and HP-beta-CD molecular dispersion resulted in more rigid molecular arrangements, which were less prone to deformation than either HP-beta-CDs or physical dispersions. The results showed how differing molecular, solid, particle, and powder state properties affect the deformation properties of the materials studied. PMID- 11028940 TI - Analysis of the compression mechanics of pharmaceutical agglomerates of different porosity and composition using the Adams and Kawakita equations. AB - PURPOSE: To analyze the mechanics of some pharmaceutical agglomerates during uniaxial confined compression by using compression parameters derived from the Heckel, Kawakita and Adams equations, and to study the influence of these compression parameters on the tablet-forming ability of agglomerates. METHODS: Force and displacement data sampled during in-die compression of agglomerates was used to calculate compression parameters according to the Heckel (sigmay), Kawakita (1/b and a), and Adams (tau0') equations. Mechanical strength of single agglomerates as well as the air permeability and tensile strength of tablets prepared from them were also determined. RESULTS: Sigmay- from the Heckel equation did not differ between agglomerates of different porosity. Both 1/b and tau0' varied with agglomerate porosity and composition. These two compression parameters were linearly related to each other. No general correlation was found between 1/b and tau0' and the strength of single agglomerates. The two parameters were related to the intergranular pore structure and tensile strength of tablets formed from the agglomerates. CONCLUSIONS: 1/b and tau0' may be interpreted as measures of the agglomerate shear strength during uniaxial confined compression, and as such they may be used as indicators of the tabletting performance of the agglomerates. PMID- 11028941 TI - Respirable PLGA microspheres containing rifampicin for the treatment of tuberculosis: manufacture and characterization. AB - PURPOSE: Particles with aerodynamic diameters of 1-5 microm deposit in the periphery of the lungs and are phagocytized by alveolar macrophages, the primary site of Mycobacterium tuberculosis infection. Aerosols of biodegradable polymeric microspheres containing antitubercular agents may be delivered to the lungs to improve the treatment of tuberculosis. METHODS: Poly(lactide-co-glycolide) (PLGA) microspheres containing rifampicin were prepared using solvent evaporation and spray drying methods. The solvent evaporation process was optimized using factorial experimental design and surface response methodology. The morphology, particle size, drug loading, and dissolution of microspheres was evaluated. RESULTS: The spray dried rifampicin loaded PLGA microparticles were shriveled, unlike the spherical particles produced by solvent evaporation. Drug loadings of 20% and 30% were achieved for solvent evaporation and spray dried products, respectively. The particles prepared by solvent evaporation and spray drying had 3.45 microm and 2.76 microm median diameters by volume, respectively. CONCLUSIONS: Respirable rifampicin loaded PLGA microspheres were produced by both solvent evaporation and spray drying methods. These particles are being evaluated in an animal model of tuberculosis. PMID- 11028943 TI - Gene transfer in vitro and in vivo by cationic lipids is not significantly affected by levels of supercoiling of a reporter plasmid. AB - PURPOSE: It is a common preconception that supercoiled plasmid DNA is more desirable for the transfection of cells that the relaxed form of the plasmid. This notion has led to the recommendation that a specification for the minimum amount of plasmid in the supercoiled form should exist in a gene therapy product. We have tested this notion by examining the effects of the degree of supercoiling on cationic lipid-mediated gene transfer in vitro and in vivo. METHODS: An ion exchange high performance liquid chromatography (HPLC) method was developed to accurately quantitate the relative amounts of supercoiled DNA in purified plasmid. A sample of the purified plasmid was fully relaxed using topoisomerase. Next, the ability of various levels of supercoiled plasmid to transfect mammalian cells was measured. RESULTS: This study suggests that there is no relation between the degree of supercoiling and lipofection efficiency. Subsequent transfection using several different lipofection agents, different cell types, and an in vivo model support these results. CONCLUSIONS: In considering a specification for the amount of supercoiled plasmid in a gene therapy product, it must be noted that the relaxed forms of the plasmid are no less efficient at gene delivery than the supercoiled forms. PMID- 11028942 TI - The effect of gelatin cross-linking on the bioequivalence of hard and soft gelatin acetaminophen capsules. AB - PURPOSE: To determine if changes in the in vitro dissolution of hard and soft gelatin acetaminophen capsules, which result from gelatin crosslinking, are predictive of changes in the bioavailability of the capsules in humans. METHODS: Both hard and soft gelatin capsules were "stressed" by a controlled exposure to formaldehyde, resulting in unstressed, moderately stressed and highly stressed capsules. In vitro dissolution studies were conducted using water or SGF with and without pepsin as the media. Separate 24-subject, 3-way crossover human bioequivalence studies were performed on the unstressed and stressed acetaminophen capsules. Plasma acetaminophen was determined by high performance liquid chromatography (HPLC) for 12 hr after each dose. RESULTS: The in vitro rate of dissolution of hard and soft gelatin capsules was decreased by crosslinking. The bioequivalence studies showed that both hard and soft gelatin capsules, which failed to meet the USP dissolution specification in water, but complied when tested in SGF containing pepsin, were bioequivalent to the unstressed control capsules. The capsules that were cross-linked to the greatest extent were not bioequivalent to the unstressed control capsules, based on Cmax. A trend toward an increase in Cmax with increased level of cross-linking was observed, but this was only significant for the severely stressed capsules. CONCLUSIONS: On the basis of this study a two-tier in vitro dissolution test was developed using enzymes to distinguish between bioequivalent and bioinequivalent gelatin capsules. PMID- 11028944 TI - Effects of pH and dose on nasal absorption of scopolamine hydrobromide in human subjects. AB - PURPOSE: The present study was conducted to evaluate the effects of formulation pH and dose on nasal absorption of scopolamine hydrobromide, the single most effective drug available for the prevention of nausea and vomiting induced by motion sickness. METHODS: Human subjects received scopolamine nasally at a dose of 0.2 mg/0.05 mL or 0.4 mg/0.10 mL, blood samples were collected at different time points, and plasma scopolamine concentrations were determined by LC-MS/MS. RESULTS: Following administration of a 0.2 mg dose, the average Cmax values were found to be 262+/-118, 419+/-161, and 488+/-331 pg/ mL for pH 4.0, 7.0, and 9.0 formulations, respectively. At the 0.4 mg dose the average Cmax values were found to be 503+/-199, 933+/-449, and 1,308+/-473 pg/mL for pH 4.0, 7.0, and 9.0 formulations, respectively. At a 0.2 mg dose, the AUC values were found to be 23,208+/-6,824, 29,145+/-9,225, and 25,721+/-5,294 pg x min/mL for formulation pH 4.0, 7.0, and 9.0, respectively. At a 0.4 mg dose, the average AUC value was found to be high for pH 9.0 formulation (70,740+/-29,381 pg x min/mL) as compared to those of pH 4.0 (59,573+/-13,700 pg x min/mL) and pH 7.0 (55,298+/-17,305 pg x min/mL) formulations. Both the Cmax and AUC values were almost doubled with doubling the dose. On the other hand, the average Tmax, values decreased linearly with a decrease in formulation pH at both doses. For example, at a 0.4 mg dose, the average Tmax values were 26.7+/-5.8, 15.0+/-10.0, and 8.8+/-2.5 minutes at formulation pH 4.0, 7.0, and 9.0, respectively. CONCLUSIONS: Nasal absorption of scopolamine hydrobromide in human subjects increased substantially with increases in formulation pH and dose. PMID- 11028945 TI - Enhancement of the systemic and CNS specific delivery of L-dopa by the nasal administration of its water soluble prodrugs. AB - PURPOSE: To study the utility of the nasal route for the systemic delivery of L dopa using water soluble prodrugs of L-dopa and to examine if this delivery method will result in preferential delivery to the CNS. METHODS: Several alkyl ester prodrugs of L-dopa were prepared and their physicochemical properties were determined. In vitro hydrolysis rate constants in buffer, rat plasma, rat brain homogenate, rat CSF, and rat nasal berfusate were determined by HPLC. In vivo nasal experiments were carried out in rats. Levels of L-dopa and dopamine in plasma, CSF, and olfactory bulb were determined using HPLC method with electrochemical detection. RESULTS: All the prodrugs showed improved solubility and lipophilicity with relatively fast in vitro conversion in rat plasma. Absorption was fast following nasal delivery of the prodrugs with bioavailability around 90%. Dopamine plasma levels did not change significantly following nasal administration of the butyl ester prodrug. Olfactory bulb and CSF L-dopa concentration were higher following nasal delivery of the butyl ester prodrug compared to an equivalent intravenous dose. CONCLUSIONS: Utilization of water soluble prodrugs of L-dopa via the nasal route in the treatment of Parkinson's disease may have therapeutic advantages such as improved bioavailability, decreased side effects, and potentially enhanced CNS delivery. PMID- 11028946 TI - Lectin-mediated drug targeting: selection of valency, sugar type (Gal/Lac), and spacer length for cluster glycosides as parameters to distinguish ligand binding to C-type asialoglycoprotein receptors and galectins. AB - PURPOSE: Common oligosaccharides of cellular glycoconjugates are ligands for more than one type of endogenous lectin. Overlapping specificities to beta galactosides of C-type lectins and galectins can reduce target selectivity of carbohydrate-ligand-dependent drug targeting. The purpose of this study is to explore distinct features of ligand presentation and structure for design of cluster glycosides to distinguish between asialoglycoprotein-specific (C-type) lectins and galectins. METHODS: Extent of binding of labeled sugar receptors to two types of matrix-immobilized (neo)glycoproteins and to cells was evaluated in the absence and presence of competitive inhibitors. This panel comprised synthetic mono-, bi-, and trivalent glycosides with two spacer lengths and galactose or lactose as ligand part. RESULTS: In contrast to C-type lectins of hepatocytes and macrophages, bi- and trivalent glycosides do not yield a notable glycoside cluster effect for galectins-1 and -3. Also, these Ca2+-independent galactoside-binding proteins prefer to home in on lactose-bearing glycosides relative to galactose as ligand, while spacer length requirements were rather similar. CONCLUSIONS: Trivalent cluster glycosides with Gal/GalNAc as ligand markedly distinguish between C-type lectins and galectins. Undesired side reactivities to galectins for C-type lectin drug delivery will thus be minimal. PMID- 11028947 TI - Anionic PAMAM dendrimers rapidly cross adult rat intestine in vitro: a potential oral delivery system? AB - PURPOSE: To investigate systematically the effect of polyamidoamine (PAMAM) dendrimer size, charge, and concentration on uptake and transport across the adult rat intestine in vitro using the everted rat intestinal sac system. METHODS: Cationic PAMAM dendrimers (generations 3 and 4) and anionic PAMAM dendrimers (generations 2.5, 3.5, and 5.5) that were modified to include on average a single pendant amino group were radioiodinated using the Bolton and Hunter Reagent. 125I-Labelled dendrimers were incubated with everted sacs in vitro and the transfer of radioactivity into the tissue and serosal fluid was followed with time. RESULTS: The serosal transfer rates seen for all anionic generations were extremely high with Endocytic Indices (EI) in the range 3.4-4.4 microL/mg protein/h. The concentration-dependence of serosal transfer was linear over the dendrimer concentration range 10-100 microg/mL. For 125I-labelled generation 5.5 the rate of tissue uptake was higher (EI = 2.48+/-0.51 microL/mg protein/h) than seen for 125I-labelled generations 2.5 and 3.5 (0.6-0.7 microL/mg protein/h) (p < 0.05). The 125I-labelled cationic PAMAM dendrimers (generations 3 and 4) displayed a tissue uptake (EI = 3.3-4.8 microL/mg protein/h) which was higher (p < 0.05) than the rate of serosal transfer (EI = 2.3-2.7 microL/mg protein/h), probably due to nonspecific adsorption of cationic dendrimer to the mucosal surface. CONCLUSIONS: As the anionic PAMAM dendrimers displayed serosal transfer rates that were faster than observed for other synthetic and natural macromolecules (including tomato lectin) studied in the everted sac system, these interesting nanoscale structures may have potential for further development as oral drug delivery systems. PMID- 11028948 TI - Inhibition of LDL oxidation and oxidized LDL-induced cytotoxicity by dihydropyridine calcium antagonists. AB - PURPOSE: The antioxidant activity of dihydropyridine calcium channel antagonists was evaluated based on LDL oxidation kinetics, oxidative cell injury associated with reactive species generation, and increases in free intracellular calcium (Ca2+) levels. Interactions with ascorbic acid were studied under conditions representative of LDL oxidation in plasma and tissue. METHODS: Analysis of antioxidant activity utilized measurements of one-electron oxidation potentials and scavenging of peroxy radical-mediated oxidation. LDL antioxidant potency was determined spectrophotometrically using copper-mediated oxidation kinetics in the absence and presence of 100 microM ascorbic acid. Prevention of oxidant-induced endothelial cell injury was determined from the formation of reactive oxygen species generation and increases in intracellular free calcium concentrations following addition of oxidized LDL or linoleic acid hydroperoxide. RESULTS: Felodipine and amlodipine effectively inhibit peroxyl radical-mediated oxidation in lipoproteins and cells that is markedly enhanced in the presence of ascorbic acid. In the presence of ascorbic acid, inhibition of LDL oxidation is over four times greater than in LDL treated without antioxidants, and oxidized LDL and linoleic acid hydroperoxide-induced reactive oxygen species formation is effectively suppressed in cells. Inhibition of intracellular calcium increases was achieved using nM concentrations of felodipine or amlodipine. CONCLUSIONS: The additive effect for ascorbic acid and the calcium channel antagonist is postulated to involve a combination of peroxide-degrading and peroxyl radical scavenging reactions, demonstrating the importance of lipid peroxides during LDL oxidation and oxidized LDL-induced cytotoxicity. Cytoprotection is associated with inhibition of oxidant-induced increases in intracellular free calcium. Both the cytoprotective and LDL antioxidant activity for these compounds is manifested at concentrations approaching the therapeutic levels found in plasma. PMID- 11028949 TI - Metabolism kinetics of beclomethasone propionate esters in human lung homogenates. AB - PURPOSE: The purposes of this study were to characterize the kinetics of beclomethasone dipropionate (BDP) and its 17-monopropionate ester (17-BMP) in human lung 1000g supernatant (HLu) at 37 degrees C, and to analyze the interindividual variability in the metabolism of BDP in HLu. METHODS: The concentrations of BDP and its metabolites were determined by HPLC with UV detection at 242 nm. Kinetics of BDP and 17-BMP decomposition were characterized by least-squares fitting of rate equations. RESULTS: The active metabolite 17-BMP was rapidly formed following the incubation of BDP in HLu. Kinetics of BDP and 17 BMP in HLu were nonlinear owing to product inhibition and enzyme saturation. A model taking into account the product inhibition provides a kinetic basis for understanding the in vivo behavior of BDP and its metabolites in human lung. There was approximately a 3.5-fold difference in the initial half-life of BDP in HLu observed in seven subjects. CONCLUSIONS: An effective activation of BDP was demonstrated in HLu through the rapid formation of 17-BMP. Kinetics of BDP and 17 BMP in HLu were well characterized by the nonlinear kinetic model. Interindividual difference in the initial half-life of BDP was due mainly to esterase metabolizing activity rather than binding affinity. PMID- 11028950 TI - Estimation of molecular linear free energy relationship descriptors. 4. Correlation and prediction of cell permeation. AB - PURPOSE: The passage of molecules across cell membranes is a crucial step in many physiological processes. We therefore seek physical models of this process, in order to predict permeation for new molecules, and to better understand the important interactions which determine the rate of permeation. METHODS: Several sets of cell permeation data reported by Collander have been correlated against calculated Linear Free Energy Relation (LFER) descriptors. These descriptors, taken as the sum of fragmental contributions, cover the size, polarity, polarizabilty, and hydrogen bonding capacity of each molecule. RESULTS: For 36 values of permeation into Chara ceratophylla cells, a model (sd = 0.24) dominated by hydrogen bond acidity is found, while for 63 rates of permeation values into Nitella cells a very similar model yields sd = 0.46. Comparisons between the two cell types are made directly for 17 compounds in both data sets, indicate differences of a similar magnitude to the standard deviations of the above models. The two data sets can be combined to yield a generic model of rates of permeation into cells, resulting in an sd value of 0.46 for a total of 100 data points. CONCLUSIONS: Models allowing accurate prediction of cell permeation have been constructed using 100 experimental data. We demonstrate that hydrogen bond acidity is the dominating factor in determining cell permeation for two distinct species of algal cell. PMID- 11028951 TI - Influence of stereoselective pharmacokinetics in the development and predictability of an IVIVC for the enantiomers of metoprolol tartrate. AB - PURPOSE: To investigate the ability of an IVIVC developed with a racemate drug as well as each enantiomer in predicting the in vivo enantiomer drug performance. METHODS: Dissolution of metoprolol extended release tablets with different release characteristics (e.g., fast (F), moderate (M), and slow (S)) was performed using USP Apparatus I, pH 1.2, 50 rpm. Metoprolol racemate tablets (S, M, and F, 100 mg) and 50 mg oral solution were administered to healthy volunteers, blood samples were collected over 24 (solution) and 48 (tablet) hours and assayed. IVIVC models developed were: (1) Racemate-fraction of drug dissolved (FRD) vs Racemate-fraction of drug absorbed (FRA), (2) R-FRD vs R-FRA, and (3) S FRD vs S-FRA for combinations of formulations (S/M/F, S/M, S/F, and M/F). Enantiomer Cmax and AUC prediction errors (PEs) were estimated for model evaluation after convolution of in vivo release rates. RESULTS: The R-IVIVC and S IVIVC accurately predicted the R- and S-metoprolol pharmacokinetic profiles, respectively. The averaged prediction errors (PE) for the enantiomer Cmax and AUC were less than 10% for S/M/F, M/F, and S/F IVIVC models. Racemate-IVIVC (M/F) was able to predict S-enantiomer with an average %PE of 2.52 for S-Cmax and 4.3 for S AUC. However, the racemate-IVIVC was unable to predict the R-enantiomer pharmacokinetic profile. CONCLUSIONS: Metoprolol racemate data cannot be used to accurately predict R-enantiomer drug concentrations. However, the racemate data was predictive of the active stereoisomer. PMID- 11028952 TI - In situ determination of partition and diffusion coefficients in the lipid bilayers of stratum corneum. PMID- 11028953 TI - Effects of storage conditions on the free volume of polyvinylpyrrolidone: comparison of positron lifetime data with the tensile strength of tablets. PMID- 11028954 TI - Staphylococcal enterotoxins. AB - Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, thus causing various types of disease symptoms. Staphylococcal enterotoxins (SEs), a family of nine major serological types of heat stable enterotoxins, are a leading cause of gastroenteritis resulting from consumption of contaminated food. In addition, SEs are powerful superantigens that stimulate non-specific T-cell proliferation. SEs share close phylogenetic relationships, with similar structures and activities. Here we review the structure and function of each known enterotoxin. PMID- 11028955 TI - Impact of environment and interspecific interactions between spoilage fungi and Aspergillus ochraceus on growth and ochratoxin production in maize grain. AB - Using layers of irradiated but still fertile maize grain, the effects of water activity (0.995, 0.95 a(w)) and temperature (18, 30 degrees C) on interspecific interactions between Aspergillus ochraceus and five other spoilage fungi were examined. Asp. ochraceus was not competitive against Asp. flavus, Asp. niger, or Alternaria alternata at 18 degrees C when water was freely available (0.995 a(w)), while at 0.95 a(w) it was dominant against Asp. candidus, Asp. flavus and Alt. alternata. At 30 degrees C and 0.995 a(w), Asp. ochraceus was dominated by other fungi, except Alt. alternata, and was mutually antagonistic to Asp. candidus and Eurotium amstelodami. However, at 30 degreees C and 0.95 a(w), it was competitive against Asp. candidus and Alt. alternata, but not against the other species examined. The overall Index of Dominance showed that Asp. ochraceus was not competitive under the conditions examined here. At 18 degrees C ochratoxin production by Asp. ochraceus was inhibited significantly by Asp. candidus (0.995 and 0.95 a(w)) and Asp. niger (0.995 a(w)). When grown on maize grain at 30 degrees C, ochratoxin production by Asp. ochraceus was significantly inhibited by other spoilage fungi when both were grown on maize grain, especially by Asp. niger and E. amstelodami (0.995 a(w)) and Asp. flavus at 0.95 a(w). These results suggest that, to a large extent, A. ochraceus is not as competitive as some other spoilage fungi in primary resource capture on maize grain at a(w) of 0.95 or above, although it may modify resource quality and influence secondary colonisation by other species under the conditions tested. PMID- 11028956 TI - Survival of bifidobacteria in yogurt and simulated gastric juice following immobilization in gellan-xanthan beads. AB - A novel acid-stable bead made of gellan gum and xanthan gum was used to immobilize Bifidobacteria. The beads (0.75% gellan and 1% xanthan gum) had an average diameter of 3 mm and did not shrink in 25% lactic acid solution, pH 1.5, or 20% acetic acid solution, pH 1.5, after storage at 4 degrees C for 4 weeks. Bifidobacterium infantis ATCC 15697, the most acid-tolerant strain tested, was immobilized in gellan-xanthan beads and its survival in peptone water, pH 4, pasteurized yogurt, and simulated gastric juice was monitored. In peptone water, pH 4, the reduction in cell count of immobilized cells of B. infantis ATCC 15697 was not significantly different from that obtained with free cells during 6 weeks of storage at 4 degrees C. However, counts of immobilized cells of B. infantis ATCC 15697 remained significantly higher than free cells (P < 0.0001) when both were exposed to simulated gastric juices at pH 2.5, 2.0 and 1.5. At pH 2.5, the viable count of free cells dropped from 1.23 x 10(9) CFU/ml to an undetectable level (< 10 CFU/ml) in 30 min, while the viable count of immobilized cells decreased by only 0.67 log cycle in the same time period. Immobilized cells also survived significantly better than free cells (P <0.05) in pasteurized yogurt after refrigerated storage for 5 weeks. PMID- 11028957 TI - Application of polynomial models to predict growth of mixed cultures of Pseudomonas spp. and Listeria in meat. AB - Three models for one rapid and one slow growing strain of Pseudomonas fragi and one slow growing strain of P. fluorescens were developed in a meat broth; they were designed to take account of variations in growth and to provide a growth response interval. These models, and another for Listeria monocytogenes (Lm14 model), were used to predict the growth of spoilage Pseudomonas spp. and pathogenic Listeria in meat products. The Pseudomonas and Listeria models provided satisfactory predictions concerning inoculated strains grown in decontaminated beef meat. It was also possible to use the Pseudomonas models to predict the growth of the natural flora (mainly Pseudomonas spp.) of refrigerated meat stored under aerobic conditions. In experiments with mixed populations, three situations were observed: (1) in decontaminated meat, L. monocytogenes inoculated alone grew well at 6 degrees C, and this result was correctly predicted by the model; (2) in decontaminated meat inoculated with Listeria and Pseudomonas strains, L. innocua grew well and was not affected by the presence of Pseudomonas, and the growth of both organisms was correctly predicted by the models; (3) in naturally contaminated meat inoculated with Listeria, the strain did not grow until Pseudomonas had reached the stationary phase. The models satisfactorily predicted the growth of Pseudomonas spp. but not that of Listeria. In conclusion, the Lm14 model cannot be used for refrigerated meat stored aerobically as the results suggest a 'fail-safe' level which may be too high: meat had already reached a spoilage state even though no increase in the level of Listeria was observed. The Pseudomonas models accurately predicted the growth of naturally occurring Pseudomonas spp. PMID- 11028958 TI - Maillard reaction causes suppression of virulence gene expression in Listeria monocytogenes. AB - Many environmental signals affect the expression of virulence genes of the food borne pathogen Listeria monocytogenes. In addition media composition has been shown to suppress levels of haemolytic activity. Using a Pr(plcA)::luxAB reporter gene fusion it was observed that the heat processing of media also reduces the level of virulence gene expression in L. monocytogenes without affecting its growth. Physicochemical factors that are considered to enhance the Maillard reaction were also found to increase the levels of suppression. The results indicate that heat treatment of a multicomponent matrix gives rise to specific inhibitors of the Listeria virulence gene operon. PMID- 11028959 TI - Estimating the annual fraction of eggs contaminated with Salmonella enteritidis in the United States. AB - Using available data on the occurrence of Salmonella enteritidis (SE) in US layer flocks and eggs, and a probabilistic scenario tree method, an estimate of the fraction of SE-contaminated eggs produced annually is derived with attendant uncertainty. In lieu of a definitive prevalence survey, the approach presented here provides insight to the relative contribution of various pathways leading to contaminated eggs. A Monte Carlo model with four branches is developed. The first branch predicts the proportion of all US flocks that are SE-affected. The second branch apportions SE-affected flocks into three categories (high, moderate, and low level affected flocks) based on population-adjusted epidemiologic data. The third branch predicts the proportion of affected flocks that are molted and producing eggs during a high risk period subsequent to molt. The fourth branch predicts the fraction of contaminated eggs produced by flocks of the type described by the pathway (e.g. high level affected flocks that are not molted) based on egg sampling evidence from naturally infected flocks. The model is simulated to account for uncertainty in the data used to estimate the branch probabilities. Correlation analysis is used to estimate the sensitivity of model output to various model inputs. The output of this model is an uncertainty distribution for the fraction of all eggs that are SE-contaminated during 1 year of production in the US. The expected value of this distribution is approximately one SE-affected egg in every 20,000 eggs annually produced, and the 90% certainty interval is between one SE-contaminated egg in 30,000 eggs, and one SE contaminated egg in 12,000 eggs. The model estimates that an average of 14% of all eggs (i.e. contaminated and not contaminated) from affected flocks are produced by high level, non-molted affected flocks, but these flocks are estimated to account for more than two-thirds of the total fraction of contaminated eggs produced annually. Sensitivity analysis also suggests that the proportion of affected flocks that are high level flocks - and the egg contamination frequency for these types of flocks - are the most sensitive model inputs. The model's pathways provide a framework for evaluating interventions to reduce the number of contaminated eggs produced in the US. Furthermore, sensitivity analysis of the model identifies those inputs whose uncertainty is most influential on the model's output. Future farm-level research priorities can be established on the basis of this analysis, but public policy decisions require a fuller exposure assessment and dose-response analysis to account for microbial growth dynamics, meal preparation, and consumption demographics among US egg consumers. PMID- 11028960 TI - Synergistic inhibition of Listeria monocytogenes on cold-smoked rainbow trout by nisin and sodium lactate. AB - The inhibition of Listeria monocytogenes and mesophilic aerobic bacteria in cold smoked rainbow trout by nisin, sodium lactate or their combination was studied. Nisin (4000-6000 IU/ml), sodium lactate (60%) or their combination (1:1) were injected into rainbow trout at an industrial scale before the smoking process, or injected into the finished smoked product. Both types of fish samples were smoked, sliced and vacuum-packed according to normal practice in the plant. Packages were opened and L. monocytogenes was inoculated (10(3)-10(4) log colony forming units (cfu)/g) onto the fish samples, which were then vacuum packed again. Samples were stored at 8 degrees C for 17 days or at 3 degrees C for 29 days. Listeria and mesophilic aerobic bacteria counts were measured once a week. The effects of treatments on sensory characteristics and storage stability were also analyzed. Both nisin and lactate inhibited the growth of L. monocytogenes in smoked fish, but the combination of the two compounds was even more effective. The combination of nisin and sodium lactate injected into smoked fish decreased the count of L. monocytogenes from 3.26 to 1.8 log cfu/g over 16 days of storage at 8 degrees C. The level of L. monocytogenes remained almost constant (4.66-4.92 log cfu/g) for 29 days at 3 degrees C in the samples injected before smoking and which contained both nisin and sodium lactate. The treatments did not affect the sensory characteristics of cold-smoked rainbow trout. Based on a triangle test, the sensory quality of all test samples remained unchanged for 23 days of storage at 3 degrees C, whereas the control fish prepared without additives or additional salt remained unchanged only for 16 days. PMID- 11028961 TI - Survival of Salmonella typhimurium and Escherichia coli O157:H7 in cheese brines. AB - Survival of Salmonella typhimurium and Escherichia coli O157:H7 was studied in model brines and brine from three cheese plants. Three strain mixtures of S. typhimurium and E. coli O157:H7 (10(6) CFU/ml) were inoculated separately into 23% model brine with or without added pasteurized whey (2%) and as a combined inoculum into the commercial brines. The model brines were incubated at 8 and 15 degrees C for 28 days, and the commercial brines at 4 and 13 degrees C for 35 days. Populations of both pathogens in the model brine + whey decreased slowly over 28 days (1.0-2.0 log CFU/ml) with greater survival at 8 degrees C than at 15 degrees C. Corresponding decreases in model brine without whey were 1.9-3.0 log CFU/ml, with greater survival at 8 degrees C than at 15 degrees C. Both S. typhimurium and E. coli O157:H7 survived significantly better (P < 0.05) at 4 degrees C than at 13 degrees C in two of the commercial brines. The survival of each pathogen in the commercial brines at 13 degrees C was significantly influenced by brine pH. Both pathogen populations decreased most rapidly in commercial brines during the first week of storage (2.5-4.0 and 2.3-2.8 log CFU/ml for S. typhimurium and E. coli O157:H7, respectively) with significant recovery (ca. 0.5 log CFU/ml increase) often occurring in the second week of storage. Counts changed little thereafter. Overall, E. coli O157:H7 survived better than S. typhimurium, with differences of 0.1-1.2 log CFU/ml between the two pathogens. Results of this study show that cheese brine could support the survival of contaminating S. typhimurium and E. coli O157:H7 for several weeks under typical brining conditions. PMID- 11028962 TI - Comparison of sodium hypochlorite and peracetic acid as sanitising agents for stainless steel food processing surfaces using epifluorescence microscopy. AB - The effects of the sanitising agents sodium hypochlorite and peracetic acid on Escherichia coli, Pseudomonas fluorescens and Staphylococcus aureus adhering to stainless steel were compared using epifluorescence microscopy. The bacteria were isolated from chicken carcasses and allowed to adhere to stainless steel coupons for 1 h before being rinsed with sterile distilled water and treated with the sanitising agents at 250 or 1000 mg l(-1) (peracetic acid) or 100 or 200 mg l(-1) (hypochlorite) for 10 min. P. fluorescens showed the greatest adhesive ability, followed by E. coli, while S. aureus adhered in lowest numbers. In all cases, sodium hypochlorite was more effective than peracetic acid in killing or removing the adherent cells. After treatment with either concentration of hypochlorite, the number of adhered cells per field (area 8.66 x 10(-3) mm2) was reduced from 118.5, 52.0 and 28.0 to 1.0, 0.0 and 0.0 for E. coli, Pseudomonas fluorescens and Staphylococcus aureus, respectively. These are equivalent to reductions from 13.7 x 10(3), 6.0 x 10(3) and 3.2 x 10(3) to 1.2 x 10(2) cells mm(-2) for E. coli and less than this number for the other two species. A median value of zero was not attained for any of the peracetic acid-treated coupons. This sanitising agent was the least effective against S. aureus, achieving only a little over 50% reduction in viable adhered cell numbers at 250 mg l(-1). In view of the importance of these microorganisms as food contaminants, and on economic grounds, peracetic acid cannot be recommended as the sanitising agent of choice for chicken processing equipment. PMID- 11028964 TI - Triumph, but self-criticism too. PMID- 11028963 TI - What hopes for GM food? PMID- 11028965 TI - Europe urged to set up advisory body on research infrastructure. PMID- 11028966 TI - Canadian science told to look north. PMID- 11028967 TI - UK faces rethink on AIDS treatment. PMID- 11028968 TI - NSF aims to inject more maths into biology. PMID- 11028969 TI - France backs IT and life sciences. PMID- 11028971 TI - Antibiotic resistance must be monitored, US Senate is told. PMID- 11028970 TI - Disagreements derail US bill to double research funding...as physical sciences plan lobbying body. PMID- 11028972 TI - Report praises European radio telescope network. PMID- 11028973 TI - Brazilian science: a springboard to success. PMID- 11028974 TI - Organic electronics: cheap and cheerful circuits. PMID- 11028975 TI - Don't dismiss chlorine; it could help us to avoid the fate of the Romans. PMID- 11028976 TI - Bright future in the stars for big telescopes? PMID- 11028978 TI - Science-based security under threat. PMID- 11028977 TI - Non-scientists thanked for vital help in Kansas. PMID- 11028979 TI - An electromagnetic personality. PMID- 11028980 TI - Through the shaving mirror. PMID- 11028981 TI - Evolving evolvability. PMID- 11028982 TI - Geochemistry. Hot heads and cold tails. PMID- 11028983 TI - Globins in the brain. PMID- 11028984 TI - Optical astronomy. The resolution revolution. PMID- 11028985 TI - The twenty-first amino acid. PMID- 11028987 TI - Use your neighbour's genes. PMID- 11028986 TI - Statistical physics. Following the crowd. PMID- 11028988 TI - Mark Oliphant (1901-2000). PMID- 11028989 TI - Maternal age and traits in offspring. PMID- 11028990 TI - Maze-solving by an amoeboid organism. PMID- 11028991 TI - RNA polymerase II elongation through chromatin. AB - The machinery that transcribes protein-coding genes in eukaryotic cells must contend with repressive chromatin structures in order to find its target DNA sequences. Diverse arrays of proteins modify the structure of chromatin at gene promoters to help transcriptional regulatory proteins access their DNA recognition sites. The way in which disruption of chromatin structure at a promoter is transmitted through a whole gene has not been defined. Recent breakthroughs suggest that the passage of an RNA polymerase through a gene is coupled to mechanisms that propagate the breakdown of chromatin. PMID- 11028992 TI - A yeast prion provides a mechanism for genetic variation and phenotypic diversity. AB - A major enigma in evolutionary biology is that new forms or functions often require the concerted effects of several independent genetic changes. It is unclear how such changes might accumulate when they are likely to be deleterious individually and be lost by selective pressure. The Saccharomyces cerevisiae prion [PSI+] is an epigenetic modifier of the fidelity of translation termination, but its impact on yeast biology has been unclear. Here we show that [PSI+] provides the means to uncover hidden genetic variation and produce new heritable phenotypes. Moreover, in each of the seven genetic backgrounds tested, the constellation of phenotypes produced was unique. We propose that the epigenetic and metastable nature of [PSI+] inheritance allows yeast cells to exploit pre-existing genetic variation to thrive in fluctuating environments. Further, the capacity of [PSI+] to convert previously neutral genetic variation to a non-neutral state may facilitate the evolution of new traits. PMID- 11028993 TI - Direct detection of pulsations of the Cepheid star zeta Gem and an independent calibration of the period-luminosity relation. AB - Cepheids are a class of variable (pulsating) stars whose absolute luminosities are related in a simple manner to their pulsational periods. By measuring the period and using the 'period-luminosity' relationship, astronomers can use the observed visual brightness to determine the distance to the star. Because these stars are very luminous, they can be observed in other galaxies, and therefore can be used to help determine the expansion rate of the Universe (the Hubble constant). Calibration of the period-luminosity relation is a necessary first step, but the small number of sufficiently nearby Cepheids has forced the use of a number of indirect means, with associated systematic uncertainties. Here we present a distance to the Cepheid zeta Geminorum, determined using a direct measurement (by an optical interferometer) of its changes in diameter as it pulsates. Within our uncertainty of 15 per cent, our distance is in agreement with previous indirect determinations. Planned improvements to the instrument will allow us to calibrate directly the period-luminosity relation to better than a few per cent. PMID- 11028994 TI - Simulating dynamical features of escape panic. AB - One of the most disastrous forms of collective human behaviour is the kind of crowd stampede induced by panic, often leading to fatalities as people are crushed or trampled. Sometimes this behaviour is triggered in life-threatening situations such as fires in crowded buildings; at other times, stampedes can arise during the rush for seats or seemingly without cause. Although engineers are finding ways to alleviate the scale of such disasters, their frequency seems to be increasing with the number and size of mass events. But systematic studies of panic behaviour and quantitative theories capable of predicting such crowd dynamics are rare. Here we use a model of pedestrian behaviour to investigate the mechanisms of (and preconditions for) panic and jamming by uncoordinated motion in crowds. Our simulations suggest practical ways to prevent dangerous crowd pressures. Moreover, we find an optimal strategy for escape from a smoke-filled room, involving a mixture of individualistic behaviour and collective 'herding' instinct. PMID- 11028995 TI - Single photons on demand from a single molecule at room temperature. AB - The generation of non-classical states of light is of fundamental scientific and technological interest. For example, 'squeezed' states enable measurements to be performed at lower noise levels than possible using classical light. Deterministic (or triggered) single-photon sources exhibit non-classical behaviour in that they emit, with a high degree of certainty, just one photon at a user-specified time. (In contrast, a classical source such as an attenuated pulsed laser emits photons according to Poisson statistics.) A deterministic source of single photons could find applications in quantum information processing, quantum cryptography and certain quantum computation problems. Here we realize a controllable source of single photons using optical pumping of a single molecule in a solid. Triggered single photons are produced at a high rate, whereas the probability of simultaneous emission of two photons is nearly zero--a useful property for secure quantum cryptography. Our approach is characterized by simplicity, room temperature operation and improved performance compared to other triggered sources of single photons. PMID- 11028996 TI - Logical computation using algorithmic self-assembly of DNA triple-crossover molecules. AB - Recent work has demonstrated the self-assembly of designed periodic two dimensional arrays composed of DNA tiles, in which the intermolecular contacts are directed by 'sticky' ends. In a mathematical context, aperiodic mosaics may be formed by the self-assembly of 'Wang' tiles, a process that emulates the operation of a Turing machine. Macroscopic self-assembly has been used to perform computations; there is also a logical equivalence between DNA sticky ends and Wang tile edges. This suggests that the self-assembly of DNA-based tiles could be used to perform DNA-based computation. Algorithmic aperiodic self-assembly requires greater fidelity than periodic self-assembly, because correct tiles must compete with partially correct tiles. Here we report a one-dimensional algorithmic self-assembly of DNA triple-crossover molecules that can be used to execute four steps of a logical (cumulative XOR) operation on a string of binary bits. PMID- 11028997 TI - Nano-sized transition-metal oxides as negative-electrode materials for lithium ion batteries. AB - Rechargeable solid-state batteries have long been considered an attractive power source for a wide variety of applications, and in particular, lithium-ion batteries are emerging as the technology of choice for portable electronics. One of the main challenges in the design of these batteries is to ensure that the electrodes maintain their integrity over many discharge-recharge cycles. Although promising electrode systems have recently been proposed, their lifespans are limited by Li-alloying agglomeration or the growth of passivation layers, which prevent the fully reversible insertion of Li ions into the negative electrodes. Here we report that electrodes made of nanoparticles of transition-metal oxides (MO, where M is Co, Ni, Cu or Fe) demonstrate electrochemical capacities of 700 mA h g(-1), with 100% capacity retention for up to 100 cycles and high recharging rates. The mechanism of Li reactivity differs from the classical Li insertion/deinsertion or Li-alloying processes, and involves the formation and decomposition of Li2O, accompanying the reduction and oxidation of metal nanoparticles (in the range 1-5 nanometres) respectively. We expect that the use of transition-metal nanoparticles to enhance surface electrochemical reactivity will lead to further improvements in the performance of lithium-ion batteries. PMID- 11028998 TI - Origins of sulphate in Antarctic dry-valley soils as deduced from anomalous 17O compositions. AB - The dry valleys of Antarctica are some of the oldest terrestrial surfaces on the Earth. Despite much study of soil weathering and development, ecosystem dynamics and the occurrence of life in these extreme environments, the reasons behind the exceptionally high salt content of the dry-valley soils have remained uncertain. In particular, the origins of sulphate are still controversial; proposed sources include wind-blown sea salt, chemical weatherings, marine incursion, hydrothermal processes and oxidation of biogenic sulphur in the atmosphere. Here we report measurements of delta18O and delta17O values of sulphates from a range of dry valley soils. These sulphates all have a large positive anomaly of 17O, of up to 3.4/1000. This suggests that Antarctic sulphate comes not just from sea salt (which has no anomaly of 17O) but also from the atmospheric oxidation of reduced gaseous sulphur compounds, the only known process that can generate the observed 17O anomaly. This source is more prominent in high inland soils, suggesting that the distributions of sulphate are largely explained by differences in particle size and transport mode which exist between sea-salt aerosols and aerosols formed from biogenic sulphur emission. PMID- 11028999 TI - Transient high temperatures in mantle plume heads inferred from magnesian olivines in Phanerozoic picrites. AB - Both scaled laboratory experiments and numerical models of terrestrial mantle plumes produce 'balloon-on-a-string' structures, with a bulbous head followed by a stem-like tail. Discussions have focused on whether their initial upwelling heads are hotter than the tails or cooler, as a result of entrainment of ambient mantle during ascent, and also on whether initial plume upwelling is a newtonian or non-newtonian process. The temperature of the mantle delivered to the base of the lithosphere is a critical parameter in such debates. Dry continental magmas can normally contribute little to this topic because their hottest (ultramafic) examples can be expected to be trapped, owing to their density, beneath the Moho. Here we report a rare case in which olivine (with 93.3% forsterite; Mg2SiO4) phenocrysts, precipitated from an unerupted komatiitic melt (approximately 24% MgO) of the Tristan mantle plume head 132 Myr ago, were carried to upper-crust levels in northwest Namibia by less Mg-rich (9.6-18.5% MgO) magmas. We infer that the hidden melt, generated when the plume impinged on the base of the lithosphere, originated in the mantle with a potential temperature of approximately 1,700 degrees C. This is approximately 400 degrees C above ambient and much hotter than the temperatures previously calculated for steady-state Phanerozoic mantle plumes. Published data show that the same conclusion can be reached for the initial Iceland and Galapagos plumes. PMID- 11029000 TI - Evolutionary instability of ectomycorrhizal symbioses in basidiomycetes. AB - Mycorrhizae, the symbiotic associations of plant roots and fungal hyphae, are classic examples of mutualisms. In these ecologically important associations, the fungi derive photosynthetic sugars from their plant hosts, which in turn benefit from fungus-mediated uptake of mineral nutrients. Early views on the evolution of symbioses suggested that all long-term, intimate associations tend to evolve toward mutualism. Following this principle, it has been suggested that mycorrhizal symbioses are the stable derivatives of ancestral antagonistic interactions involving plant parasitic fungi. Alternatively, mutualisms have been interpreted as inherently unstable reciprocal parasitisms, which can be disrupted by conflicts of interest among the partners. To determine the number of origins of mycorrhizae, and to assess their evolutionary stability, it is necessary to understand the phylogenetic relationships of the taxa involved. Here we present a broad phylogenetic analysis of mycorrhizal and free-living homobasidiomycetes (mushroom-forming fungi). Our results indicate that mycorrhizal symbionts with diverse plant hosts have evolved repeatedly from saprotrophic precursors, but also that there have been multiple reversals to a free-living condition. These findings suggest that mycorrhizae are unstable, evolutionarily dynamic associations. PMID- 11029001 TI - The genome sequence of the thermoacidophilic scavenger Thermoplasma acidophilum. AB - Thermoplasma acidophilum is a thermoacidophilic archaeon that thrives at 59 degrees C and pH 2, which was isolated from self-heating coal refuse piles and solfatara fields. Species of the genus Thermoplasma do not possess a rigid cell wall, but are only delimited by a plasma membrane. Many macromolecular assemblies from Thermoplasma, primarily proteases and chaperones, have been pivotal in elucidating the structure and function of their more complex eukaryotic homologues. Our interest in protein folding and degradation led us to seek a more complete representation of the proteins involved in these pathways by determining the genome sequence of the organism. Here we have sequenced the 1,564,905-base pair genome in just 7,855 sequencing reactions by using a new strategy. The 1,509 open reading frames identify Thermoplasma as a typical euryarchaeon with a substantial complement of bacteria-related genes; however, evidence indicates that there has been much lateral gene transfer between Thermoplasma and Sulfolobus solfataricus, a phylogenetically distant crenarchaeon inhabiting the same environment. At least 252 open reading frames, including a complete protein degradation pathway and various transport proteins, resemble Sulfolobus proteins most closely. PMID- 11029002 TI - An SNP map of the human genome generated by reduced representation shotgun sequencing. AB - Most genomic variation is attributable to single nucleotide polymorphisms (SNPs), which therefore offer the highest resolution for tracking disease genes and population history. It has been proposed that a dense map of 30,000-500,000 SNPs can be used to scan the human genome for haplotypes associated with common diseases. Here we describe a simple but powerful method, called reduced representation shotgun (RRS) sequencing, for creating SNP maps. RRS re-samples specific subsets of the genome from several individuals, and compares the resulting sequences using a highly accurate SNP detection algorithm. The method can be extended by alignment to available genome sequence, increasing the yield of SNPs and providing map positions. These methods are being used by The SNP Consortium, an international collaboration of academic centres, pharmaceutical companies and a private foundation, to discover and release at least 300,000 human SNPs. We have discovered 47,172 human SNPs by RRS, and in total the Consortium has identified 148,459 SNPs. More broadly, RRS facilitates the rapid, inexpensive construction of SNP maps in biomedically and agriculturally important species. SNPs discovered by RRS also offer unique advantages for large-scale genotyping. PMID- 11029003 TI - An SNP map of human chromosome 22. AB - The human genome sequence will provide a reference for measuring DNA sequence variation in human populations. Sequence variants are responsible for the genetic component of individuality, including complex characteristics such as disease susceptibility and drug response. Most sequence variants are single nucleotide polymorphisms (SNPs), where two alternate bases occur at one position. Comparison of any two genomes reveals around 1 SNP per kilobase. A sufficiently dense map of SNPs would allow the detection of sequence variants responsible for particular characteristics on the basis that they are associated with a specific SNP allele. Here we have evaluated large-scale sequencing approaches to obtaining SNPs, and have constructed a map of 2,730 SNPs on human chromosome 22. Most of the SNPs are within 25 kilobases of a transcribed exon, and are valuable for association studies. We have scaled up the process, detecting over 65,000 SNPs in the genome as part of The SNP Consortium programme, which is on target to build a map of 1 SNP every 5 kilobases that is integrated with the human genome sequence and that is freely available in the public domain. PMID- 11029004 TI - A vertebrate globin expressed in the brain. AB - Haemoglobins and myoglobins constitute related protein families that function in oxygen transport and storage in humans and other vertebrates. Here we report the identification of a third globin type in man and mouse. This protein is predominantly expressed in the brain, and therefore we have called it neuroglobin. Mouse neuroglobin is a monomer with a high oxygen affinity (half saturation pressure, P50 approximately 2 torr). Analogous to myoglobin, neuroglobin may increase the availability of oxygen to brain tissue. The human neuroglobin gene (NGB), located on chromosome 14q24, has a unique exon-intron structure. Neuroglobin represents a distinct protein family that diverged early in metazoan evolution, probably before the Protostomia/Deuterostomia split. PMID- 11029005 TI - Immune control of HIV-1 after early treatment of acute infection. AB - Virus-specific T-helper cells are considered critical for the control of chronic viral infections. Successful treatment of acute HIV-1 infection leads to augmentation of these responses, but whether this enhances immune control has not been determined. We administered one or two supervised treatment interruptions to eight subjects with treated acute infection, with the plan to restart therapy if viral load exceeded 5,000 copies of HIV-1 RNA per millilitre of plasma (the level at which therapy has been typically recommended) for three consecutive weeks, or 50,000 RNA copies per ml at one time. Here we show that, despite rebound in viraemia, all subjects were able to achieve at least a transient steady state off therapy with viral load below 5,000 RNA copies per ml. At present, five out of eight subjects remain off therapy with viral loads of less than 500 RNA copies per ml plasma after a median 6.5 months (range 5-8.7 months). We observed increased virus-specific cytotoxic T lymphocytes and maintained T-helper-cell responses in all. Our data indicate that functional immune responses can be augmented in a chronic viral infection, and provide rationale for immunotherapy in HIV-1 infection. PMID- 11029006 TI - arrow encodes an LDL-receptor-related protein essential for Wingless signalling. AB - The Wnt family of secreted molecules functions in cell-fate determination and morphogenesis during development in both vertebrates and invertebrates (reviewed in ref. 1). Drosophila Wingless is a founding member of this family, and many components of its signal transduction cascade have been identified, including the Frizzled class of receptor. But the mechanism by which the Wingless signal is received and transduced across the membrane is not completely understood. Here we describe a gene that is necessary for all Wingless signalling events in Drosophila. We show that arrow gene function is essential in cells receiving Wingless input and that it acts upstream of Dishevelled. arrow encodes a single pass transmembrane protein, indicating that it may be part of a receptor complex with Frizzled class proteins. Arrow is a low-density lipoprotein (LDL)-receptor related protein (LRP), strikingly homologous to murine and human LRP5 and LRP6. Thus, our data suggests a new and conserved function for this LRP subfamily in Wingless/Wnt signal reception. PMID- 11029007 TI - LDL-receptor-related proteins in Wnt signal transduction. AB - The Wnt family of secreted signalling molecules are essential in embryo development and tumour formation. The Frizzled (Fz) family of serpentine receptors function as Wnt receptors, but how Fz proteins transduce signalling is not understood. In Drosophila, arrow phenocopies the wingless (DWnt-1) phenotype, and encodes a transmembrane protein that is homologous to two members of the mammalian low-density lipoprotein receptor (LDLR)-related protein (LRP) family, LRP5 and LRP6 (refs 12-15). Here we report that LRP6 functions as a co-receptor for Wnt signal transduction. In Xenopus embryos, LRP6 activated Wnt-Fz signalling, and induced Wnt responsive genes, dorsal axis duplication and neural crest formation. An LRP6 mutant lacking the carboxyl intracellular domain blocked signalling by Wnt or Wnt-Fz, but not by Dishevelled or beta-catenin, and inhibited neural crest development. The extracellular domain of LRP6 bound Wnt-1 and associated with Fz in a Wnt-dependent manner. Our results indicate that LRP6 may be a component of the Wnt receptor complex. PMID- 11029008 TI - An LDL-receptor-related protein mediates Wnt signalling in mice. AB - Wnt genes comprise a large family of secreted polypeptides that are expressed in spatially and tissue-restricted patterns during vertebrate embryonic development. Mutational analysis in mice has shown the importance of Wnts in controlling diverse developmental processes such as patterning of the body axis, central nervous system and limbs, and the regulation of inductive events during organogenesis. Although many components of the Wnt signalling pathway have been identified, little is known about how Wnts and their cognate Frizzled receptors signal to downstream effector molecules. Here we present evidence that a new member of the low-density lipoprotein (LDL)-receptor-related protein family, LRP6 (ref. 3), is critical for Wnt signalling in mice. Embryos homozygous for an insertion mutation in the LRP6 gene exhibit developmental defects that are a striking composite of those caused by mutations in individual Wnt genes. Furthermore, we show a genetic enhancement of a Wnt mutant phenotype in mice lacking one functional copy of LRP6. Together, our results support a broad role for LRP6 in the transduction of several Wnt signals in mammals. PMID- 11029010 TI - Effect of tea and other dietary factors on iron absorption. AB - Iron deficiency is a major world health problem, that is, to a great extent, caused by poor iron absorption from the diet. Several dietary factors can influence this absorption. Absorption enhancing factors are ascorbic acid and meat, fish and poultry; inhibiting factors are plant components in vegetables, tea and coffee (e.g., polyphenols, phytates), and calcium. After identifying these factors their individual impact on iron absorption is described. Specific attention was paid to the effects of tea on iron absorption. We propose a calculation model that predicts iron absorption from a meal. Using this model we calculated the iron absorption from daily menus with varying amounts of enhancers and inhibitors. From these calculations we conclude that the presence of sufficient amounts of iron absorption enhancers (ascorbic acid, meat, fish, poultry, as present in most industrialized countries) overcomes inhibition of iron absorption from even large amounts of tea. In individuals with low intakes of heme iron, low intakes of enhancing factors and/or high intakes of inhibitors, iron absorption may be an issue. Depletion of iron stores enhances iron absorption, but this effect is not adequate to compensate for the inhibition of iron absorption in such an inadequate dietary situation. For subjects at risk of iron deficiency, the following recommendations are made. Increase heme-iron intake (this form of dietary iron present in meat fish and poultry is hardly influenced by other dietary factors with respect to its absorption); increase meal-time ascorbic acid intake; fortify foods with iron. Recommendations with respect to tea consumption (when in a critical group) include: consume tea between meals instead of during the meal; simultaneously consume ascorbic acid and/or meat, fish and poultry. PMID- 11029009 TI - Interaction of oestrogen receptor with the regulatory subunit of phosphatidylinositol-3-OH kinase. AB - Oestrogen produces diverse biological effects through binding to the oestrogen receptor (ER). The ER is a steroid hormone nuclear receptor, which, when bound to oestrogen, modulates the transcriptional activity of target genes. Controversy exists, however, concerning whether ER has a role outside the nucleus, particularly in mediating the cardiovascular protective effects of oestrogen. Here we show that the ER isoform, ER alpha, binds in a ligand-dependent manner to the p85alpha regulatory subunit of phosphatidylinositol-3-OH kinase (PI(3)K). Stimulation with oestrogen increases ER alpha-associated PI(3)K activity, leading to the activation of protein kinase B/Akt and endothelial nitric oxide synthase (eNOS). Recruitment and activation of PI(3)K by ligand-bound ER alpha are independent of gene transcription, do not involve phosphotyrosine adapter molecules or src-homology domains of p85alpha, and extend to other steroid hormone receptors. Mice treated with oestrogen show increased eNOS activity and decreased vascular leukocyte accumulation after ischaemia and reperfusion injury. This vascular protective effect of oestrogen was abolished in the presence of PI(3)K or eNOS inhibitors. Our findings define a physiologically important non nuclear oestrogen-signalling pathway involving the direct interaction of ER alpha with PI(3)K. PMID- 11029011 TI - Control of foodborne pathogens during sufu fermentation and aging. AB - Control of the foodborne pathogens Escherichia coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes during sufu fermentation was evaluated. Before fermentation, pathogens were inoculated onto tofu (substrate for sufu) at 5 log cfu/g or 3 log cfu/g, and starter culture (Actinomucor elegans) was inoculated at 3 log cfu/g. After 2 days of fermentation at 30 degrees C, the four pathogens reached 7 to 9 log cfu/g, and the mold count reached 6 to 7 log cfu/g. After fermentation, sufu samples were aged in a solution of 10% alcohol + 12% NaCl. After 1 month of aging, the total bacterial count was 6 to 7 log cfu/g, but all foodborne pathogens and mold were reduced to nondetectable levels. The total bacterial count decreased after aging for 2 months and 3 months, but the differences were not significant (P > 0.05) compared with the count after 1 month. Microorganism in experimental sufu from different aging periods and in commercial sufu were compared. A total of 270 isolates were purified and identified by the BBL Crystal Identification System. From the experimental sufu samples, 49 Bacillus spp. (20.4%), 167 Enterococcus spp. (69.6%), 6 Shewanella putrefaciens (2.4%), and 18 miscellaneous Gram-negative bacilli (7.5%) were identified. From commercial sufu samples, 17 Bacillus spp. (56.7%), 2 Enterococcus durans (6.7%), 5 miscellaneous Gram-negative bacilli (16.7%), 5 Corynbacterium aquaticum (16.7%), and 1 Shewanella putrefaciens (3.3%) were obtained. Although the longer aging period did not significantly decrease the total bacterial count, it may help in the development of sufu flavor. This study showed that sufu fermentation and aging can control common foodborne pathogens, so sufu is a safe product even though its preparation does not include pasteurization. PMID- 11029012 TI - Low-fat/calorie foods: current state and perspectives. AB - Low-fat/calorie products were originally developed for diabetics and people with specific health problems and they were considerably expensive. Nowadays, consumers' demand for low-fat/calorie products has significantly raised in an attempt to limit health problems, to lose or stabilize their weight, and to work within the frame of a healthier diet. The food industry has been confronted with a new challenge in order to satisfy consumers; development of low-fat/calories products with acceptable sensory characteristics and competitive price, by preferably employing the conventional processing equipment and in agreement with current strict legislation. The role of fat replacers and sugar substitutes in the successful manufacture of these products is crucial. PMID- 11029013 TI - Is fatigue in patients with multiple sclerosis related to autonomic dysfunction? AB - Time-dependent frequency decomposition of fluctuations in cardiovascular signals (heart rate [HR], blood pressure, and blood flow) provides noninvasive and quantitative evaluation of autonomic activity during transient and steady-state conditions. This method was applied during a change of position from supine to standing in patients with multiple sclerosis (MS) who experienced unexplained fatigue and in age-matched control subjects. No difference in response to standing, as reflected in the time domain parameters (mean HR, mean blood pressure, and mean blood flow), was observed between patients with MS and control subjects. Moreover, no difference was observed in very-low-frequency and low frequency (related to sympathetic activity) content of HR, blood pressure, blood flow, or high-frequency content of HR (related to parasympathetic activity). The only spectral estimates that showed a significant difference between groups were the ratio of low-frequency to high-frequency content of HR and low-frequency content of HR normalized to total power. Both these parameters provide an estimate of the sympathovagal balance. A significant increase in these two estimates on standing was observed in control subjects only, indicating possible impairment of the sympathovagal balance response to standing in patients with MS who experienced fatigue. The authors observed a significant age dependence between close age subgroups, which occurred in the MS group only and was observed in some of the investigated spectral estimates that reflect vagal activity. Therefore, the authors assumed that age-related reduction in vagal activity occurred earlier in patients with MS who experienced fatigue. This reduction could also explain the lack of increase in the sympathovagal balance on standing. To validate this enhanced age dependence, further investigation should be performed in a larger group of subjects with a wider age range. PMID- 11029014 TI - Quantitative sensory testing of thermal and vibratory perception in familial dysautonomia. AB - Familial dysautonomia (FD) is an inherited disorder that is known to affect both sensory and autonomic functions as a result of incomplete neuronal development and progressive loss but the degree to which patients are affected differs greatly. To determine if quantitative vibration and thermal testing refined the assessment of severity, 23 familial dysautonomia patients were evaluated by clinical examination, measurements of median, peroneal and sural nerve conduction velocities (NCV), and assessment of vibration thresholds at two body sites and of warm and cold perception thresholds at 6 body sites using the method of limits. Data from 80 age-matched normal individuals provided control data for vibration and temperature thresholds. All familial dysautonomia patients had abnormal thermal thresholds. Vibration perception was abnormal in 20 patients. NCVs were slowed in 8 of 16 patients who agreed to be tested. Abnormalities in thermal thresholds are consistent with the reduction of small nerve fibers in familial dysautonomia Abnormal vibration thresholds might be due to disturbed conduction of vibratory impulse trains and reflect the degree to which the disorder is progressive. Vibration and thermal sensation testing were better accepted and provided more information than NCV regarding severity of disease. PMID- 11029015 TI - Detection of serious bradyarrhythmias in Guillain-Barre syndrome: sensitivity and specificity of the 24-hour heart rate power spectrum. AB - This study was undertaken to determine the nature of bradyarrhythmic events and their relationship to motor disability, disease stages and tachycardia in patients with Guillain-Barre syndrome, and to investigate the potential of the 24 hour heart rate power spectrum (HRPS) for the detection of serious bradyarrhythmias in individual patients. Thirty-five consecutive patients with Guillain-Barre syndrome who were admitted to the authors' intensive care unit were studied. In all patients, the heart rate was continuously recorded during the early stages of the disease, averaged at 1-minute intervals, and stored for 1 to 87 days. The HRPS (n = 556, 16 +/- 19 per patient; median, 9) was calculated by Fourier analysis of 24-hour recordings and logarithmically transformed. The slope was estimated by regression analysis of log (power) on log (frequency) between 10(-4) and 4 x 10(-3) Hz, showing an inverse power law behavior in all 556 HRPSs. Eleven patients (31%) had serious bradyarrhythmias. Most of these patients were not dependent on mechanical ventilation, with 3 of 11 patients (27%) still being able to walk more than 5 meters. Sustained tachycardia occurred less frequently in patients with than in those without bradyarrhythmias. The combination of the slope of the power law regression line and the log (power) at 10(-4) Hz (log P4) of the 24-hour HRPS correctly identified 8 of 11 bradyarrhythmic patients (sensitivity 73%) and 16 of 22 patients with Guillain Barre syndrome who did not have bradyarrhythmias (specificity 73%). All bradyarrhythmic patients could be detected in the subgroup of patients without sustained tachycardia. The 24-hour HRPS is a powerful predictor of serious autonomic complications in patients with Guillain-Barre syndrome and may help to identify patients at risk of potentially life-threatening arrhythmias. PMID- 11029016 TI - The use of clonidine in the management of autonomic overactivity in neuroleptic malignant syndrome. AB - The aim of this study was to identify the effectiveness of clonidine in the recovery of patients with neuroleptic malignant syndrome and autonomic dysfunction, including blood pressure lability. Nine patients with neuroleptic malignant syndrome and autonomic dysfunction were treated with clonidine in the intensive care unit, according to a protocol, and the results were compared with those of seven patients with the same syndrome who were not treated with clonidine. Clonidine was administered until blood pressure stability was fully restored, and thereafter the dose was gradually reduced. There was a significant reduction in the duration of ventilation and stay in the intensive care unit in the clonidine group. Three patients from the nonclonidine group died. The data suggest that in the clonidine group, patients with neuroleptic malignant syndrome and autonomic dysfunction appear to have better and faster recovery, especially in blood pressure control, after intravenous clonidine treatment. PMID- 11029018 TI - Prader-Willi syndrome fails to alter cardiac autonomic modulation. AB - Twenty-six healthy subjects with a diagnosis of Prader-Willi syndrome were compared with 26 age-, gender-, and body mass index-matched controls for autonomic modulation and baroreflex sensitivity. Electrocardiograms, beat-to-beat finger blood pressures, and respiration were recorded for several minutes in the following sequence: (1) supine, (2) after transition from supine to standing, (3) sitting, (4) during a Valsalva maneuver, (5) while performing moderate exercise, and (6) during recovery from exercise while seated. All recordings were channeled and stored in a computer; analyses were carried out at a later date. Power spectral analysis (fast-Fourier transform) of heart period variability was used to assess cardiac autonomic modulation. The slope of the regression equation between heart period and blood pressure rise after the Valsalva maneuver was used as an index of baroreflex sensitivity. Analysis of variance failed to reveal significant differences in any of the autonomic and baroreflex sensitivity variables between the two groups. Because breathing patterns entrain autonomic modulation, we verified respiration and found no differences between the two groups. Therefore, findings in the current investigation indicate that cardiac autonomic modulation in patients with Prader-Willi syndrome does not differ from age and body mass index-matched subjects. PMID- 11029017 TI - Acetylsalicylic acid and autonomic modulation. AB - Loss of autonomic balance characterized by increased sympathetic activity and decreased vagal activity has been implicated as a major cardiovascular risk factor. Aspirin's cardioprotective abilities involve a multitude of physiologic processes. However, the effects of aspirin on cardiac autonomic activity are unknown. In a double-blind crossover study, 22 subjects randomly received either aspirin or placebo in the amounts of 325 mg with each meal (three times per day) over a 2.5-day period. The total amount of aspirin ingested was 2,275 mg, which resulted in plasma levels of 3.3 mg/dl. At the conclusion of each treatment, subjects were evaluated for autonomic physiology activity using standard autonomic tests. Power spectral analyses of the electrocardiograms were used to delineate autonomic function. A 2 x 4 repeated measures analysis of variance revealed significant and favorable changes in autonomic activity after the use of aspirin. Specifically, at rest high-frequency (HF) power was significantly higher (mean, 1,090 + 1,463.5 msec2) compared with the placebo (mean, 692 742 msec2) (p <0.05). Low-frequency (LF) power was significantly reduced (mean, 963 745 msec2) after aspirin compared with placebo (mean, 1,100 906 msec2). After the aspirin treatment, a significantly lower LF-to-HF power ratio (mean, 1.7 2 msec2) was noted at rest when compared with the placebo (mean, 2.5 2.7 msec2) (p <0.05). Similar significant trends were seen during the sustained isometric contraction after aspirin therapy for HF power (mean 210 2.15 msec2) compared with placebo (mean, 213 184 msec2) (p <0.05). Accordingly, the LF-to-HF power ratio was lower as well when compared to placebo treatment (mean, 2.3 3.5 msec2) (mean, 5.3 8.4 msec2) (p <0.05). No differences were found in breathing rates for hemodynamic variables between any of the protocols. The significant reduction of LF-to-HF ratio, a marker of sympathovagal balance, for both protocols appeared to be largely due to a withdrawal of LF modulation and concomitant but lesser increase in HF modulation. Favorable alterations in autonomic outflow through prostaglandin inhibition may be one of the mechanisms by which low therapeutic amounts of aspirin provide prophylactic cardioprotection. PMID- 11029019 TI - Sympathetic nervous system activity and cardiovascular homeostatis during head-up tilt in patients with spinal cord injuries. AB - The relationship between sympathetic nervous system activity and cardiovascular responses to head-up tilt in patients with spinal cord injuries and in able bodied subjects was studied. Twenty-seven adults, nine in each of the three groups (tetraplegia, paraplegia, and able-bodied subjects) were tilted 70 degrees, head up, for 12 minutes after 20 minutes supine rest. Differences between steady-state measurements of mean arterial pressure, stroke volume, and sympathetic nervous system activity were estimated in both positions. Sympathetic nervous system activity was reflected by the low-frequency peak of the blood pressure variability spectrum. From supine rest to head-up tilt, low-frequency power increased in able-bodied subjects (median, 0.42 mm Hg2, p = 0.003), which was different (p = 0.015) from patients with tetraplegia and paraplegia (-0.15 and -0.10 mm Hg2, respectively). Stroke volume and mean arterial pressure decreased in patients with tetraplegia (-40% and -9 mm Hg, respectively; p = 0.008, both variables) more than in able-bodied subjects (-33%, 11 mm Hg, respectively) or patients with paraplegia (-24%, 8 mm Hg, respectively). Results indicated increased sympathetic nervous system activity during head-up tilt in able-bodied subjects, but not in patients with paraplegia or tetraplegia, whereas patients with tetraplegia, but not paraplegia, showed poorer cardiovascular homeostasis than able-bodied subjects. This suggests that patients with paraplegia maintained cardiovascular homeostasis during head-up tilt without increased sympathetic nervous system activity. PMID- 11029020 TI - Timeout postponement without increased reinforcement frequency. AB - Three experiments were conducted to examine pigeons' postponement of signaled extinction periods (timeouts) from a schedule of food reinforcement when such responding neither decreased overall timeout frequency nor increased the overall frequency of food reinforcement. A discrete-trial procedure was used in which a response during the first 5 s of a trial postponed an otherwise immediate 60-s timeout to a later part of that same trial but had no effect on whether the timeout occurred. During time-in periods, responses on a second key produced food according to a random-interval 20-s schedule. In Experiment 1, the response timeout interval was 45 s under postponement conditions and 0 s under extinction conditions (responses were ineffective in postponing timeouts). The percentage of trials with a response was consistently high when the timeout-postponement contingency was in effect and decreased to low levels when it was discontinued under extinction conditions. In Experiment 2, the response-timeout interval was also 45 s but postponement responses increased the duration of the timeout, which varied from 60 s to 105 s across conditions. Postponement responding was maintained, generally at high levels, at all timeout durations, despite sometimes large decreases in the overall frequency of food reinforcement. In Experiment 3, timeout duration was held constant at 60 s while the response-timeout interval was varied systematically across conditions from 0 s to 45 s. Postponement responding was maintained under all conditions in which the response-timeout interval exceeded 0 s (the timeout interval in the absence of a response). In some conditions of Experiment 3, which were designed to control for the immediacy of food reinforcement and food-correlated (time-in) stimuli, responding postponed timeout but the timeout was delayed whether a response occurred or not. Responding was maintained for 2 of 3 subjects, suggesting that behavior was negatively reinforced by timeout postponement rather than positively reinforced by the more immediate presentation of food or food-correlated (time-in) stimuli. PMID- 11029021 TI - Comparing preference and resistance to change in constant- and variable-duration schedule components. AB - Two experiments explored preference and resistance to change in concurrent chains in which the terminal links were variable-interval schedules that ended either after a single reinforcer had been delivered (variable duration) or after a fixed period of access to the schedule (constant duration). In Experiment 1, pigeons' preference between the same pair of terminal links overmatched relative reinforcement rate when the terminal links were of constant duration, but not when they were of variable duration. Responding during the richer terminal link decreased less, relative to baseline, when response-independent food was presented during the initial links according to a variable-time schedule. In Experiment 2, all subjects consistently preferred a terminal link that consisted of 20-s access to a variable-interval 20-s schedule over a terminal link that ended after one reinforcer had been delivered by the same schedule. Results of resistance-to-change tests corresponded to preference, as responding during the constant-duration terminal link decreased less, relative to baseline, when disrupted by both response-independent food during the initial links and prefeeding. Overall, these data extend the general covariation of preference and resistance to change seen in previous studies. However, they suggest that reinforcement numerosity, including variability in the number of reinforcers per terminal-link entry, may sometimes affect preference and resistance to change in ways that are difficult to explain in terms of current models. PMID- 11029022 TI - Synthesizing concurrent interval performances. AB - Concurrent schedules may be viewed as consisting of two pairs of stay and switch schedules, each pair associated with one of the alternatives. A stay schedule arranges reinforcers for staying and responding at one alternative, whereas the associated switch schedule arranges reinforcers for switching to the other alternative. In standard concurrent schedules, the stay schedule at each alternative is equivalent to the switch schedule at the other alternative. MacDonall (1999) exposed rats to one pair of stay and switch variable-ratio schedules and varied the response requirements across conditions. Combining results from symmetric pairs produced composite performances that were described by the generalized matching law. This outcome was noteworthy because the data were obtained from performances at two alternatives with contingencies that were functionally unrelated to each other. This result suggests that concurrent performances may consist of two unrelated performances that alternate as behavior moves between alternatives. The purpose of the present experiment was to extend those results to interval schedules. Rats were exposed to pairs of random interval schedules, and across conditions their mean intervals were varied. When data from appropriately paired conditions were combined, the composite performances were consistent with the generalized matching law. In addition, the results supported two models of concurrent performances that were based on local variables at an alternative (behavior, and stay and switch reinforcers): a modified version of the contingency discrimination model (Davison & Jenkins, 1985) and the local model (MacDonall, 1999). PMID- 11029023 TI - Derived relational responding as generalized operant behavior. AB - The major aim of the present study was to demonstrate that derived relational responding may be viewed as a form of generalized operant behavior. In Experiment 1, 4 subjects were divided into two conditions (2 in each condition). Using a two comparison matching-to-sample procedure, all subjects were trained and tested for the formation of two combinatorially entailed relations. Subjects were trained and tested across multiple stimulus sets. Each set was composed of novel stimuli. Both Conditions 1 and 2 involved explicit performance-contingent feedback presented at the end of each block of test trials (i.e., delayed feedback). In Condition 1, feedback was accurate (consistent with the experimenter-designated relations) following exposure to the initial stimulus sets. When subjects' responding reached a predefined mastery criterion, the feedback then switched to inaccurate (not consistent with the experimenter-designated relations) until responding once again reached a predefined criterion. Condition 2 was similar to Condition 1, except that exposure to the initial stimulus sets was followed by inaccurate feedback and once the criterion was reached feedback switched to accurate. Once relational responding emerged and stabilized, response patterns on novel stimulus sets were controlled by the feedback delivered for previous stimulus sets. Experiment 2 replicated Experiment 1, except that during Conditions 3 and 4 four comparison stimuli were employed during training and testing. Experiment 3 was similar to Condition 1 of Experiment 1, except that after the mastery criterion was reached for class-consistent responding, feedback alternated from accurate to inaccurate across each successive stimulus set. Experiment 4 involved two types of feedback, one type following tests for mutual entailment and the other type following tests for combinatorial entailment. Results from this experiment demonstrated that mutual and combinatorial entailment may be controlled independently by accurate and inaccurate feedback. Overall, the data support the suggestion, made by relational frame theory, that derived relational responding is a form of generalized operant behavior. PMID- 11029024 TI - The effects of morphine on fixed-interval patterning and temporal discrimination. AB - Changes produced by drugs in response patterns under fixed-interval schedules of reinforcement have been interpreted to result from changes in temporal discrimination. To examine this possibility, this experiment determined the effects of morphine on the response patterning of 4 pigeons during a fixed interval 1-min schedule of food delivery with interpolated temporal discrimination trials. Twenty of the 50 total intervals were interrupted by choice trials. Pecks to one key color produced food if the interval was interrupted after a short time (after 2 or 4.64 s). Pecks to another key color produced food if the interval was interrupted after a long time (after 24.99 or 58 s). Morphine (1.0 to 10.0 mg/kg) decreased the index of curvature (a measure of response patterning) during fixed intervals and accuracy during temporal discrimination trials. Accuracy was equally disrupted following short and long sample durations. Although morphine disrupted temporal discrimination in the context of a fixed-interval schedule, these effects are inconsistent with interpretations of the disruption of response patterning as a selective overestimation of elapsed time. The effects of morphine may be related to the effects of more conventional external stimuli on response patterning. PMID- 11029025 TI - Differential outcome effect in the horse. AB - Three horses were trained with a discrimination task in which the color (blue or yellow) of a center panel signaled the correct (left or right) response (lever press). Reinforcing outcomes for the two correct color-position combinations (blue-left and yellow-right) were varied across phases. Discrimination performance was better when the combinations were differentially reinforced by two types of food (chopped carrot pieces and a solid food pellet) than when the combinations were randomly reinforced by these outcomes or when there was a common reinforcer for each of the correct combinations. However, the discrimination performance established by the differential outcome procedure was still 80% to 90% correct, and an analysis of two-trial sequences revealed that the stimulus color of the preceding trial interfered with discrimination performance on a given trial. Our demonstration of the differential outcome effect in the horse and its further analysis might contribute to more efficient control of equine behavior in the laboratory as well as in horse sports. PMID- 11029026 TI - The philosophical terrain of behavior analysis: a review of B. A. Thyer (Ed.), The Philosophical Legacy of Behaviorism. AB - The Philosophical Legacy of Behaviorism, edited by Bruce A. Thyer, is a set of original contributions, each dealing, from a behavioral stance, with one of the following major topics of philosophy: epistemology, ethics, consciousness, language, free will and determinism, and self-control. Confusions about radical behaviorism and its similarities to, and differences from, other behavioral and non-behavioral approaches are described in the book, which provides a state-of the-art description of the philosophical underpinnings of behavior analysis. PMID- 11029027 TI - Colony architectonics and heterogeneity of Escherichia coli populations. PMID- 11029028 TI - Magnetoselection effects in EPR spectra of the Primary-Donor Triplet State in Reaction Centers of the phototrophic bacterium Rhodobacter sphaeroides R26. PMID- 11029030 TI - Paradoxical wave acceleration in the sink-type boundary of an excitable medium. PMID- 11029029 TI - The SHF dielectric permeability of polyhydroxybutyrate, a degradable biological polymer. PMID- 11029031 TI - A possible mechanism of the nonexponential electron transfer in DNA. PMID- 11029032 TI - Effect of high-power microwave radiation with nanosecond pulse duration on some biological objects. PMID- 11029033 TI - Potassium distribution in the Neurospora crassa hypha. PMID- 11029034 TI - Protection of telomeres by the Ku protein in fission yeast. AB - Schizosaccharomyces pombe cells survive loss of telomeres by a unique pathway of chromosome circularization. Factors potentially involved in this survival mechanism include the heterodimeric Ku protein and ligase IV, both of which are involved in the repair of DNA double-strand breaks in mammalian cells. Furthermore, Ku plays a role in telomere maintenance as well as in DNA double strand break repair in Saccharomyces cerevisiae. We have identified Ku and ligase IV homologues in S. pombe and analyzed their functions during normal growth and in cells undergoing senescence. In the absence of either a Ku subunit (pku70(+)) or ligase IV (lig4(+)), nonhomologous DNA end-joining was severely reduced. Lack of functional Ku led to shorter but stable telomeres and caused striking rearrangements of telomere-associated sequences, indicating a function for Ku in inhibiting recombinational activities near chromosome ends. In contrast to S. cerevisiae, concurrent deletion of pku70(+) and the gene for the catalytic subunit of telomerase (trt1(+)) was not lethal, allowing for the first time the dissection of the roles of Ku during senescence. Our results support a model in which Ku protects chromosome termini from nucleolytic and recombinational activities but is not involved in the formation of chromosome end fusions during senescence. The conclusion that nonhomologous end-joining is not required for chromosome circularization was further supported by analysis of survivors in strains lacking the genes for both trt1(+) and lig4(+). PMID- 11029035 TI - The diabetes autoantigen ICA69 and its Caenorhabditis elegans homologue, ric-19, are conserved regulators of neuroendocrine secretion. AB - ICA69 is a diabetes autoantigen with no homologue of known function. Given that most diabetes autoantigens are associated with neuroendocrine secretory vesicles, we sought to determine if this is also the case for ICA69 and whether this protein participates in the process of neuroendocrine secretion. Western blot analysis of ICA69 tissue distribution in the mouse revealed a correlation between expression levels and secretory activity, with the highest expression levels in brain, pancreas, and stomach mucosa. Subcellular fractionation of mouse brain revealed that although most of the ICA69 pool is cytosolic and soluble, a subpopulation is membrane-bound and coenriched with synaptic vesicles. We used immunostaining in the HIT insulin-secreting beta-cell line to show that ICA69 localizes in a punctate manner distinct from the insulin granules, suggesting an association with the synaptic-like microvesicles found in these cells. To pursue functional studies on ICA69, we chose to use the model organism Caenorhabditis elegans, for which a homologue of ICA69 exists. We show that the promoter of the C. elegans ICA69 homologue is specifically expressed in all neurons and specialized secretory cells. A deletion mutant was isolated and found to exhibit resistance to the drug aldicarb (an inhibitor of acetylcholinesterase), suggesting defective neurotransmitter secretion in the mutant. On the basis of the aldicarb resistance phenotype, we named the gene ric-19 (resistance to inhibitors of cholinesterase-19). The resistance to aldicarb was rescued by introducing a ric-19 transgene into the ric-19 mutant background. This is the first study aimed at dissecting ICA69 function, and our results are consistent with the interpretation that ICA69/RIC-19 is an evolutionarily conserved cytosolic protein participating in the process of neuroendocrine secretion via association with certain secretory vesicles. PMID- 11029036 TI - The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes. AB - Endobrevin/VAMP-8 is an R-SNARE localized to endosomes, but it is unknown in which intracellular fusion step it operates. Using subcellular fractionation and quantitative immunogold electron microscopy, we found that endobrevin/VAMP-8 is present on all membranes known to communicate with early endosomes, including the plasma membrane, clathrin-coated pits, late endosomes, and membranes of the trans Golgi network. Affinity-purified antibodies that block the ability of endobrevin/VAMP-8 to form SNARE core complexes potently inhibit homotypic fusion of both early and late endosomes in vitro. Fab fragments were as active as intact immunoglobulin Gs. Recombinant endobrevin/VAMP-8 inhibited both fusion reactions with similar potency. We conclude that endobrevin/VAMP-8 operates as an R-SNARE in the homotypic fusion of early and late endosomes. PMID- 11029037 TI - Ajuba, a cytosolic LIM protein, shuttles into the nucleus and affects embryonal cell proliferation and fate decisions. AB - Cellular adhesive events affect cell proliferation and differentiation decisions. How cell surface events mediating adhesion transduce signals to the nucleus is not well understood. After cell-cell or cell-substratum contact, cytosolic proteins are recruited to clustered adhesion receptor complexes. One such family of cytosolic proteins found at sites of cell adhesion is the Zyxin family of LIM proteins. Here we demonstrate that the family member Ajuba was recruited to the cell surface of embryonal cells, upon aggregate formation, at sites of cell-cell contact. Ajuba contained a functional nuclear export signal and shuttled into the nucleus. Importantly, accumulation of the LIM domains of Ajuba in the nucleus of P19 embryonal cells resulted in growth inhibition and spontaneous endodermal differentiation. The differentiating effect of Ajuba mapped to the third LIM domain, whereas regulation of proliferation mapped to the first and second LIM domains. Ajuba-induced endodermal differentiation of these cells correlated with the capacity to activate c-Jun kinase and required c-Jun kinase activation. These results suggest that the cytosolic LIM protein Ajuba may provide a new mechanism to transduce signals from sites of cell adhesion to the nucleus, regulating cell growth and differentiation decisions during early development. PMID- 11029038 TI - Forced expression of keratin 16 alters the adhesion, differentiation, and migration of mouse skin keratinocytes. AB - Injury to the skin results in an induction of keratins K6, K16, and K17 concomitant with activation of keratinocytes for reepithelialization. Forced expression of human K16 in skin epithelia of transgenic mice causes a phenotype that mimics several aspects of keratinocyte activation. Two types of transgenic keratinocytes, with forced expression of either human K16 or a K16-C14 chimeric cDNA, were analyzed in primary culture to assess the impact of K16 expression at a cellular level. High K16-C14-expressing and low K16-expressing transgenic keratinocytes behave similar to wild type in all aspects tested. In contrast, high K16-expressing transgenic keratinocytes show alterations in plating efficiency and calcium-induced differentiation, but proliferate normally. Migration of keratinocytes is reduced in K16 transgenic skin explants compared with controls. Finally, a subset of high K16-expressing transgenic keratinocytes develops major changes in the organization of keratin filaments in a time- and calcium concentration-dependent manner. These changes coincide with alterations in keratin content while the steady-state levels of K16 protein remain stable. We conclude that forced expression of K16 in progenitor skin keratinocytes directly impacts properties such as adhesion, differentiation, and migration, and that these effects depend upon determinants contained within its carboxy terminus. PMID- 11029039 TI - Polymerization defects within human telomerase are distinct from telomerase RNA and TEP1 binding. AB - The minimal, active core of human telomerase is postulated to contain two components, the telomerase RNA hTER and the telomerase reverse transcriptase hTERT. The reconstitution of human telomerase activity in vitro has facilitated the identification of sequences within the telomerase RNA and the RT motifs of hTERT that are essential for telomerase activity. However, the precise role of residues outside the RT domain of hTERT is unknown. Here we have delineated several regions within hTERT that are important for telomerase catalysis, primer use, and interaction with the telomerase RNA and the telomerase-associated protein TEP1. In particular, certain deletions of the amino and carboxy terminus of hTERT that retained an interaction with telomerase RNA and TEP1 were nonetheless completely inactive in vitro and in vivo. Furthermore, hTERT truncations lacking the amino terminus that were competent to bind the telomerase RNA were severely compromised for the ability to elongate telomeric and nontelomeric primers. These results suggest that the interaction of telomerase RNA with hTERT can be functionally uncoupled from polymerization, and that there are regions outside the RT domain of hTERT that are critical for telomerase activity and primer use. These results establish that the human telomerase RT possesses unique polymerization determinants that distinguish it from other RTs. PMID- 11029040 TI - Regulation of macropinocytosis by p21-activated kinase-1. AB - The process of macropinocytosis is an essential aspect of normal cell function, contributing to both growth and motile processes of cells. p21-activated kinases (PAKs) are targets for activated Rac and Cdc42 guanosine 5'-triphosphatases and have been shown to regulate the actin-myosin cytoskeleton. In fibroblasts PAK1 localizes to areas of membrane ruffling, as well as to amiloride-sensitive pinocytic vesicles. Expression of a PAK1 kinase autoinhibitory domain blocked both platelet-derived growth factor- and RacQ61L-stimulated uptake of 70-kDa dextran particles, whereas an inactive version of this domain did not, indicating that PAK kinase activity is required for normal growth factor-induced macropinocytosis. The mechanisms by which PAK modulate macropinocytosis were examined in NIH3T3 cell lines expressing various PAK1 constructs under the control of a tetracycline-responsive transactivator. Cells expressing PAK1 (H83,86L), a mutant that dramatically stimulates formation of dorsal membrane ruffles, exhibited increased macropinocytic uptake of 70-kDa dextran particles in the absence of additional stimulation. This effect was not antagonized by coexpression of dominant-negative Rac1-T17N. In the presence of platelet-derived growth factor, both PAK1 (H83,86L) and a highly kinase active PAK1 (T423E) mutant dramatically enhanced the uptake of 70-kDa dextran. Neither wild-type PAK1 nor vector controls exhibited enhanced macropinocytosis, nor did PAK1 (H83,86L) affect clathrin-dependent endocytic mechanisms. Active versions of PAK1 enhanced both growth factor-stimulated 70-kDa dextran uptake and efflux, suggesting that PAK1 activity modulated pinocytic vesicle cycling. These data indicate that PAK1 plays an important regulatory role in the process of macropinocytosis, perhaps related to the requirement for PAK in directed cell motility. PMID- 11029041 TI - Matricellular proteins as modulators of cell-matrix interactions: adhesive defect in thrombospondin 2-null fibroblasts is a consequence of increased levels of matrix metalloproteinase-2. AB - Thrombospondin 2 (TSP2)-null mice, generated by disruption of the Thbs2 gene, display a variety of connective tissue abnormalities, including fragile skin and the presence of abnormally large collagen fibrils with irregular contours in skin and tendon. In this study we demonstrate that TSP2-null skin fibroblasts show a defect in attachment to a number of matrix proteins, and a reduction in cell spreading. To investigate the molecular mechanisms responsible for these abnormal cell-matrix interactions, we compared the levels of matrix metalloproteinases (MMPs) in wild-type and mutant fibroblasts. Isolation and analysis of gelatinases from conditioned media by gelatin-agarose affinity chromatography and gelatinolytic assays demonstrated that TSP2-null fibroblasts produce a 2-fold increase in gelatinase A (MMP2) compared with wild-type cells. The adhesive defect was corrected by treatment of TSP2-null fibroblasts with soluble TSP2, with the MMP inhibitors BB94 and tissue inhibitor of metalloproteinase-2, and with a neutralizing antibody to MMP2. Moreover, stable transfection of TSP2-null fibroblasts with mouse TSP2 cDNA corrected both the adhesive defect and the altered expression of MMP2. Finally, MMP2 was shown to interact with TSP2 in a direct-binding plate assay. We conclude that TSP2 plays an important role in cell matrix interactions, and that a deficiency in the protein results in increased levels of MMP2 that contribute to the adhesive defect in TSP2-null fibroblasts and could play a role in the complex phenotype of TSP2-null mice. PMID- 11029042 TI - The Doa4 deubiquitinating enzyme is functionally linked to the vacuolar protein sorting and endocytic pathways. AB - The Saccharomyces cerevisiae DOA4 gene encodes a deubiquitinating enzyme that is required for rapid degradation of ubiquitin-proteasome pathway substrates. Both genetic and biochemical data suggest that Doa4 acts in this pathway by facilitating ubiquitin recycling from ubiquitinated intermediates targeted to the proteasome. Here we describe the isolation of 12 spontaneous extragenic suppressors of the doa4-1 mutation; these involve seven different genes, six of which were cloned. Surprisingly, all of the cloned DID (Doa4-independent degradation) genes encode components of the vacuolar protein-sorting (Vps) pathway. In particular, all are class E Vps factors, which function in the maturation of a late endosome/prevacuolar compartment into multivesicular bodies that then fuse with the vacuole. Four of the six Did proteins are structurally related, suggesting an overlap in function. In wild-type and several vps strains, Doa4-green fluorescent protein displays a cytoplasmic/nuclear distribution. However, in cells lacking the Vps4/Did6 ATPase, a large fraction of Doa4-green fluorescent protein, like several other Vps factors, concentrates at the late endosome-like class E compartment adjacent to the vacuole. These results suggest an unanticipated connection between protein deubiquitination and endomembrane protein trafficking in which Doa4 acts at the late endosome/prevacuolar compartment to recover ubiquitin from ubiquitinated membrane proteins en route to the vacuole. PMID- 11029043 TI - Identification of a new vertebrate nucleoporin, Nup188, with the use of a novel organelle trap assay. AB - The study of the nuclear pore in vertebrates would benefit from a strategy to directly identify new nucleoporins and interactions between those nucleoporins. We have developed a novel two-step "organelle trap" assay involving affinity selection and in vitro pore assembly. In the first step, soluble proteins derived from Xenopus egg extracts are applied to a column containing a ligand of interest. The bound proteins are then tagged by biotinylation and eluted. In the second step, potential nucleoporins are selected for by virtue of their ability to assemble into annulate lamellae, a cytoplasmic mimic of nuclear pores. The incorporated proteins are then recognized by their biotin tag. Here we use the lectin wheat germ agglutinin (WGA) as ligand; WGA inhibits nuclear transport and has been shown to directly bind three known nucleoporins from Xenopus extract, Nup62, Nup98, and Nup214, all of which contain N-acetylglucosamine residues. Under reduced-stringency conditions, three additional proteins bind to WGA Sepharose and are revealed by the organelle trap assay. We identified all three as partner nucleoporins. Two were discovered to be Xenopus Nup93 and Nup205. The third is a novel vertebrate nucleoporin, Nup188. This new vertebrate protein, Xenopus Nup188, exists in a complex with xNup93 and xNup205. The Nup93-Nup188 Nup205 complex does not bind directly to WGA but binds indirectly via the N acetylglucosamine-modified nucleoporins. A gene encoding human Nup188 was also identified. The discovery of vertebrate Nup188, related to a yeast nucleoporin, and its novel protein-protein interactions illustrates the power of the two-step organelle trap assay and identifies new building blocks for constructing the nuclear pore. PMID- 11029044 TI - Cbl-transforming variants trigger a cascade of molecular alterations that lead to epithelial mesenchymal conversion. AB - Dispersal of epithelial cells is an important aspect of tumorigenesis, and invasion. Factors such as hepatocyte growth factor induce the breakdown of cell junctions and promote cell spreading and the dispersal of colonies of epithelial cells, providing a model system to investigate the biochemical signals that regulate these events. Multiple signaling proteins are phosphorylated in epithelial cells during hepatocyte growth factor-induced cell dispersal, including c-Cbl, a protooncogene docking protein with ubiquitin ligase activity. We have examined the role of c-Cbl and a transforming variant (70z-Cbl) in epithelial cell dispersal. We show that the expression of 70z-Cbl in Madin-Darby canine kidney epithelial cells resulted in the breakdown of cell-cell contacts and alterations in cell morphology characteristic of epithelial-mesenchymal transition. Structure-function studies revealed that the amino-terminal portion of c-Cbl, which corresponds to the Cbl phosphotyrosine-binding/Src homology domain 2, is sufficient to promote the morphological changes in cell shape. Moreover, a point mutation at Gly-306 abrogates the ability of the Cbl Src homology domain 2 to induce these morphological changes. Our results identify a role for Cbl in the regulation of epithelial-mesenchymal transition, including loss of adherens junctions, cell spreading, and the initiation of cell dispersal. PMID- 11029045 TI - A role for the START gene-specific transcription factor complex in the inactivation of cyclin B and Cut2 destruction. AB - Hyperactivation of Cdc2 in fission yeast causes cells to undergo a lethal premature mitosis called mitotic catastrophe. This phenotype is observed in cdc2 3w wee1-50 cells at high temperature. Eleven of 17 mutants that suppress this phenotype define a single complementation group, mcs1. The mcs1-77 mutant also suppresses lethal inactivation of the Wee1 and Mik1 tyrosine kinases and thus delays mitosis independently of Cdc2 tyrosine phosphorylation. We have cloned mcs1 by isolating suppressors of the cell cycle arrest phenotype of mcs1-77 cdc25 22 cells and found that it encodes Res2, a component of the START gene-specific transcription factor complex MBF (also known as DSC-1). The mcs1-77 mutant bears a single point mutation in the DNA-binding domain of Res2 that causes glycine 68 to be replaced by a serine residue. Importantly, two substrates of the anaphase promoting complex (APC), the major B-type cyclin, Cdc13, and the anaphase inhibitor, Cut2, are unstable in G2-phase mcs1-77 cells. Consistent with this, we observe abnormal sister chromatid separation in mcs1-77 cdc25-22 cells at the restrictive temperature. Mutation of either Cdc10 or Res1 also deregulates MBF dependent transcription and causes a G2 delay. We find that this cell cycle delay is abolished in the absence of the APC regulator Ste9/Srw1 and that the periodic expression of Ste9/Srw1 is controlled by the MBF complex. These data suggest that in fission yeast the MBF complex plays a key role in the inactivation of cyclin B and Cut2 destruction by controlling the periodic production of APC regulators. PMID- 11029046 TI - Proteasomal proteomics: identification of nucleotide-sensitive proteasome interacting proteins by mass spectrometric analysis of affinity-purified proteasomes. AB - Ubiquitin-dependent proteolysis is catalyzed by the 26S proteasome, a dynamic complex of 32 different proteins whose mode of assembly and mechanism of action are poorly understood, in part due to the difficulties encountered in purifying the intact complex. Here we describe a one-step affinity method for purifying intact 26S proteasomes, 19S regulatory caps, and 20S core particles from budding yeast cells. Affinity-purified 26S proteasomes hydrolyze both model peptides and the ubiquitinated Cdk inhibitor Sic1. Affinity purifications performed in the absence of ATP or presence of the poorly hydrolyzable analog ATP-gamma-S unexpectedly revealed that a large number of proteins, including subunits of the skp1-cullin-F-box protein ligase (SCF) and anaphase-promoting complex (APC) ubiquitin ligases, copurify with the 19S cap. To identify these proteasome interacting proteins, we used a recently developed method that enables the direct analysis of the composition of large protein complexes (DALPC) by mass spectrometry. Using DALPC, we identified more than 24 putative proteasome interacting proteins, including Ylr421c (Daq1), which we demonstrate to be a new subunit of the budding yeast 19S cap, and Ygr232w (Nas6), which is homologous to a subunit of the mammalian 19S cap (PA700 complex). Additional PIPs include the heat shock proteins Hsp70 and Hsp82, the deubiquitinating enzyme Ubp6, and proteins involved in transcriptional control, mitosis, tubulin assembly, RNA metabolism, and signal transduction. Our data demonstrate that nucleotide hydrolysis modulates the association of many proteins with the 26S proteasome, and validate DALPC as a powerful tool for rapidly identifying stoichiometric and substoichiometric components of large protein assemblies. PMID- 11029047 TI - Expression of multiple UNC-13 proteins in the Caenorhabditis elegans nervous system. AB - The Caenorhabditis elegans UNC-13 protein and its mammalian homologues are important for normal neurotransmitter release. We have identified a set of transcripts from the unc-13 locus in C. elegans resulting from alternative splicing and apparent alternative promoters. These transcripts encode proteins that are identical in their C-terminal regions but that vary in their N-terminal regions. The most abundant protein form is localized to most or all synapses. We have analyzed the sequence alterations, immunostaining patterns, and behavioral phenotypes of 31 independent unc-13 alleles. Many of these mutations are transcript-specific; their phenotypes suggest that the different UNC-13 forms have different cellular functions. We have also isolated a deletion allele that is predicted to disrupt all UNC-13 protein products; animals homozygous for this null allele are able to complete embryogenesis and hatch, but they die as paralyzed first-stage larvae. Transgenic expression of the entire gene rescues the behavior of mutants fully; transgenic overexpression of one of the transcripts can partially compensate for the genetic loss of another. This finding suggests some degree of functional overlap of the different protein products. PMID- 11029048 TI - Constitutive macropinocytosis in oncogene-transformed fibroblasts depends on sequential permanent activation of phosphoinositide 3-kinase and phospholipase C. AB - Macropinocytosis results from the closure of lamellipodia generated by membrane ruffling, thereby reflecting cortical actin dynamics. Both transformation of Rat 1 fibroblasts by v-Src or K-Ras and stable transfection for expression of dominant-positive, wild-type phosphoinositide 3-kinase (PI3K) regulatory subunit p85 alpha constitutively led to stress fiber disruption, cortical actin recruitment, extensive ruffling, and macropinosome formation, as measured by a selective acceleration of fluid-phase endocytosis. These alterations closely correlated with activation of PI3K and phosphatidylinositol-specific phospholipase C (PI-PLC), as assayed by 3-phosphoinositide synthesis in situ and in vitro and inositol 1, 4,5 trisphosphate steady-state levels, respectively; they were abolished by stable transfection of v-Src-transformed cells for dominant-negative truncated p85 alpha expression and by pharmacological inhibitors of PI3K and PI-PLC, indicating a requirement for both enzymes. Whereas PI3K activation resisted PI-PLC inhibition, PI-PLC activation was abolished by a PI3K inhibitor and dominant-negative transfection, thus placing PI-PLC downstream of PI3K. Together, these data suggest that permanent sequential activation of both PI3K and PI-PLC is necessary for the dramatic reorganization of the actin cytoskeleton in oncogene-transformed fibroblasts, resulting in constitutive ruffling and macropinocytosis. PMID- 11029049 TI - Identification of a novel saturable endoplasmic reticulum localization mechanism mediated by the C-terminus of a Dictyostelium protein disulfide isomerase. AB - Localization of soluble endoplasmic reticulum (ER) resident proteins is likely achieved by the complementary action of retrieval and retention mechanisms. Whereas the machinery involving the H/KDEL and related retrieval signals in targeting escapees back to the ER is well characterized, other mechanisms including retention are still poorly understood. We have identified a protein disulfide isomerase (Dd-PDI) lacking the HDEL retrieval signal normally found at the C terminus of ER residents in Dictyostelium discoideum. Here we demonstrate that its 57 residue C-terminal domain is necessary for intracellular retention of Dd-PDI and sufficient to localize a green fluorescent protein (GFP) chimera to the ER, especially to the nuclear envelope. Dd-PDI and GFP-PDI57 are recovered in similar cation-dependent complexes. The overexpression of GFP-PDI57 leads to disruption of endogenous PDI complexes and induces the secretion of PDI, whereas overexpression of a GFP-HDEL chimera induces the secretion of endogenous calreticulin, revealing the presence of two independent and saturable mechanisms. Finally, low-level expression of Dd-PDI but not of PDI truncated of its 57 C terminal residues complements the otherwise lethal yeast TRG1/PDI1 null mutation, demonstrating functional disulfide isomerase activity and ER localization. Altogether, these results indicate that the PDI57 peptide contains ER localization determinants recognized by a conserved machinery present in D. discoideum and Saccharomyces cerevisiae. PMID- 11029050 TI - Vimentin filaments in fibroblasts are a reservoir for SNAP23, a component of the membrane fusion machinery. AB - Soluble N-ethyl maleimide-sensitive fusion protein attachment protein receptors (SNAREs) are core machinery for membrane fusion during intracellular vesicular transport. Synaptosome-associated protein of 23 kDa (SNAP23) is a target SNARE previously identified at the plasma membrane, where it is involved in exocytotic membrane fusion. Here we show that SNAP23 associates with vimentin filaments in a Triton X-100 insoluble fraction in fibroblasts in primary culture and HeLa cells. Upon treatment of human fibroblasts with N-ethyl-maleimide, SNAP23 dissociates from vimentin filaments and forms a protein complex with syntaxin 4, a plasma membrane SNARE. The vimentin-associated pool of SNAP23 can therefore be a reservoir, which would supply the plasma membrane fusion machinery, in fibroblasts. Our observation points to a yet unexplored role of intermediate filaments. PMID- 11029051 TI - Bidirectional translocation of neurofilaments along microtubules mediated in part by dynein/dynactin. AB - Neuronal cytoskeletal elements such as neurofilaments, F-actin, and microtubules are actively translocated by an as yet unidentified mechanism. This report describes a novel interaction between neurofilaments and microtubule motor proteins that mediates the translocation of neurofilaments along microtubules in vitro. Native neurofilaments purified from spinal cord are transported along microtubules at rates of 100-1000 nm/s to both plus and minus ends. This motion requires ATP and is partially inhibited by vanadate, consistent with the activity of neurofilament-bound molecular motors. Motility is in part mediated by the dynein/dynactin motor complex and several kinesin-like proteins. This reconstituted motile system suggests how slow net movement of cytoskeletal polymers may be achieved by alternating activities of fast microtubule motors. PMID- 11029052 TI - Overexpression of beta-catenin induces apoptosis independent of its transactivation function with LEF-1 or the involvement of major G1 cell cycle regulators. AB - beta-Catenin promotes epithelial architecture by forming cell surface complexes with E-cadherin and also interacts with TCF/LEF-1 in the nucleus to control gene expression. By DNA transfection, we overexpressed beta-catenin and/or LEF-1 in NIH 3T3 fibroblasts, corneal fibroblasts, corneal epithelia, uveal melanoma cells, and several carcinoma cell lines. In all cases (with or without LEF-1), the abundant exogenous beta-catenin localizes to the nucleus and forms distinct nuclear aggregates that are not associated with DNA. Surprisingly, we found that with time (5-8 d after transfection) cells overexpressing beta-catenin all undergo apoptosis. LEF-1 does not need to be present. Moreover, LEF-1 overexpression in the absence of exogenous beta-catenin does not induce apoptosis, even though some endogenous beta-catenin moves with the exogenous LEF 1 into the nucleus. TOPFLASH/FOPFLASH reporter assays showed that full-length beta-catenin is able to induce LEF-1-dependent transactivation, whereas Arm beta catenin totally abolishes the transactivating function. However, Arm beta catenin, containing deletions of known LEF-1-transactivating domains, has the same apoptotic effects as full-length beta-catenin. Overexpressed beta-catenin also induces apoptosis in cells transfected with nuclear localization signal deleted LEF-1 that localizes only in the cytoplasm. Thus, the apoptotic effects of overexpressed exogenous beta-catenin do not rely on its transactivating function with nuclear LEF-1. Overexpressed delta-catenin, containing 10 Arm repeats, induces only minor apoptosis, suggesting that the major apoptotic effect may be due to domains specific to beta-catenin as well as to Arm repeats. The absence of p53, Rb, cyclin D1, or E2F1 does not affect the apoptotic effect of overexpressed beta-catenin, but Bcl-x(L) reduces it. We hypothesize that in vivo apoptosis of cells overexpressing beta-catenin might be a physiological mechanism to eliminate them from the population. PMID- 11029053 TI - Mps1p regulates meiotic spindle pole body duplication in addition to having novel roles during sporulation. AB - Sporulation in yeast requires that a modified form of chromosome segregation be coupled to the development of a specialized cell type, a process akin to gametogenesis. Mps1p is a dual-specificity protein kinase essential for spindle pole body (SPB) duplication and required for the spindle assembly checkpoint in mitotically dividing cells. Four conditional mutant alleles of MPS1 disrupt sporulation, producing two distinct phenotypic classes. Class I alleles of mps1 prevent SPB duplication at the restrictive temperature without affecting premeiotic DNA synthesis and recombination. Class II MPS1 alleles progress through both meiotic divisions in 30-50% of the population, but the asci are incapable of forming mature spores. Although mutations in many other genes block spore wall formation, the cells produce viable haploid progeny, whereas mps1 class II spores are unable to germinate. We have used fluorescently marked chromosomes to demonstrate that mps1 mutant cells have a dramatically increased frequency of chromosome missegregation, suggesting that loss of viability is due to a defect in spindle function. Overall, our cytological data suggest that MPS1 is required for meiotic SPB duplication, chromosome segregation, and spore wall formation. PMID- 11029054 TI - Coiled-coil trigger motifs in the 1B and 2B rod domain segments are required for the stability of keratin intermediate filaments. AB - Many alpha-helical proteins that form two-chain coiled coils possess a 13-residue trigger motif that seems to be required for the stability of the coiled coil. However, as currently defined, the motif is absent from intermediate filament (IF) protein chains, which nevertheless form segmented two-chain coiled coils. In the present work, we have searched for and identified two regions in IF chains that are essential for the stability necessary for the formation of coiled-coil molecules and thus may function as trigger motifs. We made a series of point substitutions with the keratin 5/keratin 14 IF system. Combinations of the wild type and mutant chains were assembled in vitro and in vivo, and the stabilities of two-chain (one-molecule) and two-molecule assemblies were examined with use of a urea disassembly assay. Our new data document that there is a region located between residues 100 and 113 of the 2B rod domain segment that is absolutely required for molecular stability and IF assembly. This potential trigger motif differs slightly from the consensus in having an Asp residue at position 4 (instead of a Glu) and a Thr residue at position 9 (instead of a charged residue), but there is an absolute requirement for a Glu residue at position 6. Because these 13 residues are highly conserved, it seems possible that this motif functions in all IF chains. Likewise, by testing keratin IF with substitutions in both chains, we identified a second potential trigger motif between residues 79 and 91 of the 1B rod domain segment, which may also be conserved in all IF chains. However, we were unable to find a trigger motif in the 1A rod domain segment. In addition, many other point substitutions had little detectable effect on IF assembly, except for the conserved Lys-23 residue of the 2B rod domain segment. Cross-linking and modeling studies revealed that Lys-23 may lie very close to Glu-106 when two molecules are aligned in the A(22) mode. Thus, the Glu 106 residue may have a dual role in IF structure: it may participate in trigger formation to afford special stability to the two-chain coiled-coil molecule, and it may participate in stabilization of the two-molecule hierarchical stage of IF structure. PMID- 11029055 TI - E3-13.7 integral membrane proteins encoded by human adenoviruses alter epidermal growth factor receptor trafficking by interacting directly with receptors in early endosomes. AB - Animal cell viruses provide valuable model systems for studying many normal cellular processes, including membrane protein sorting. The focus of this study is an integral membrane protein encoded by the E3 transcription region of human adenoviruses called E3-13.7, which diverts recycling EGF receptors to lysosomes without increasing the rate of receptor internalization or intrinsic receptor tyrosine kinase activity. Although E3-13.7 can be found on the plasma membrane when it is overexpressed, its effect on EGF receptor trafficking suggests that the plasma membrane is not its primary site of action. Using cell fractionation and immunocytochemical experimental approaches, we now report that the viral protein is located predominantly in early endosomes and limiting membranes of endosome-to-lysosome transport intermediates called multivesicular endosomes. We also demonstrate that E3-13.7 physically associates with EGF receptors undergoing E3-13.7-mediated down-regulation in early endosomes. Receptor-viral protein complexes then dissociate, and EGF receptors proceed to lysosomes, where they are degraded, while E3-13.7 is retained in endosomes. We conclude that E3-13.7 is a resident early endocytic protein independent of EGF receptor expression, because it has identical intracellular localization in mouse cells lacking endogenous receptors and cells expressing a human cytomegalovirus-driven receptor cDNA. Finally, we demonstrate that EGF receptor residues 675-697 are required for E3 13.7-mediated down-regulation. Interestingly, this sequence includes a known EGF receptor leucine-based lysosomal sorting signal used during ligand-induced trafficking, which is also conserved in the viral protein. E3-13.7, therefore, provides a novel model system for determining the molecular basis of selective membrane protein transport in the endocytic pathway. Our studies also suggest new paradigms for understanding EGF receptor sorting in endosomes and adenovirus pathogenesis. PMID- 11029056 TI - Phosphorylation-dependent localization of microtubule-associated protein MAP2c to the actin cytoskeleton. AB - Microtubule-associated protein 2 (MAP2) is a neuronal phosphoprotein that promotes net microtubule growth and actin cross-linking and bundling in vitro. Little is known about MAP2 regulation or its interaction with the cytoskeleton in vivo. Here we investigate the in vivo function of three specific sites of phosphorylation on MAP2. cAMP-dependent protein kinase activity disrupts the MAP2 microtubule interaction in living HeLa cells and promotes MAP2c localization to peripheral membrane ruffles enriched in actin. cAMP-dependent protein kinase phosphorylates serines within three KXGS motifs, one within each tubulin-binding repeat. These highly conserved motifs are also found in homologous proteins tau and MAP4. Phosphorylation at two of these sites was detected in brain tissue. Constitutive phosphorylation at these sites was mimicked by single, double, and triple mutations to glutamic acid. Biochemical and microscopy-based assays indicated that mutation of a single residue was adequate to disrupt the MAP2 microtubule interaction in HeLa cells. Double or triple point mutation promoted MAP2c localization to the actin cytoskeleton. Specific association between MAP2c and the actin cytoskeleton was demonstrated by retention of MAP2c-actin colocalization after detergent extraction. Specific phosphorylation states may enhance the interaction of MAP2 with the actin cytoskeleton, thereby providing a regulated mechanism for MAP2 function within distinct cytoskeletal domains. PMID- 11029057 TI - Rotational coupling of the transmembrane and kinase domains of the Neu receptor tyrosine kinase. AB - Ligand binding to receptor tyrosine kinases (RTKs) regulates receptor dimerization and activation of the kinase domain. To examine the role of the transmembrane domain in regulation of RTK activation, we have exploited a simplified transmembrane motif, [VVVEVVV](n), previously shown to activate the Neu receptor. Here we demonstrate rotational linkage of the transmembrane domain with the kinase domain, as evidenced by a periodic activation of Neu as the dimerization motif is shifted across the transmembrane domain. These results indicate that activation requires a specific orientation of the kinase domains with respect to each other. Results obtained with platelet-derived growth factor receptor-beta suggest that this rotational linkage of the transmembrane domain to the kinase domain may be a general feature of RTKs. These observations suggest that activating mutations in RTK transmembrane and juxtamembrane domains will be limited to those residues that position the kinase domains in an allowed rotational conformation. PMID- 11029059 TI - Conditional expression of a truncated fragment of nonmuscle myosin II-A alters cell shape but not cytokinesis in HeLa cells. AB - A truncated fragment of the nonmuscle myosin II-A heavy chain (NMHC II-A) lacking amino acids 1-591, delta N592, was used to examine the cellular functions of this protein. Green fluorescent protein (GFP) was fused to the amino terminus of full length human NMHC II-A, NMHC II-B, and delta N592 and the fusion proteins were stably expressed in HeLa cells by using a conditional expression system requiring absence of doxycycline. The HeLa cell line studied normally expressed only NMHC II-A and not NMHC II-B protein. Confocal microscopy indicated that the GFP fusion proteins of full-length NMHC II-A, II-B, and delta N592 were localized to stress fibers. However, in vitro assays showed that baculovirus-expressed delta N592 did not bind to actin, suggesting that delta N592 was localized to actin stress fibers through incorporation into endogenous myosin filaments. There was no evidence for the formation of heterodimers between the full-length endogenous nonmuscle myosin and truncated nonmuscle MHCs. Expression of delta N592, but not full-length NMHC II-A or NMHC II-B, induced cell rounding with rearrangement of actin filaments and disappearance of focal adhesions. These cells returned to their normal morphology when expression of delta N592 was repressed by addition of doxycycline. We also show that GFP-tagged full-length NMHC II-A or II-B, but not delta N592, were localized to the cytokinetic ring during mitosis, indicating that, in vertebrates, the amino-terminus part of mammalian nonmuscle myosin II may be necessary for localization to the cytokinetic ring. PMID- 11029058 TI - Role for the silencing protein Dot1 in meiotic checkpoint control. AB - During the meiotic cell cycle, a surveillance mechanism called the "pachytene checkpoint" ensures proper chromosome segregation by preventing meiotic progression when recombination and chromosome synapsis are defective. The silencing protein Dot1 (also known as Pch1) is required for checkpoint-mediated pachytene arrest of the zip1 and dmc1 mutants of Saccharomyces cerevisiae. In the absence of DOT1, the zip1 and dmc1 mutants inappropriately progress through meiosis, generating inviable meiotic products. Other components of the pachytene checkpoint include the nucleolar protein Pch2 and the heterochromatin component Sir2. In dot1, disruption of the checkpoint correlates with the loss of concentration of Pch2 and Sir2 in the nucleolus. In addition to its checkpoint function, Dot1 blocks the repair of meiotic double-strand breaks by a Rad54 dependent pathway of recombination between sister chromatids. In vegetative cells, mutation of DOT1 results in delocalization of Sir3 from telomeres, accounting for the impaired telomeric silencing in dot1. PMID- 11029060 TI - Yeast exocytic v-SNAREs confer endocytosis. AB - In yeast, homologues of the synaptobrevin/VAMP family of v-SNAREs (Snc1 and Snc2) confer the docking and fusion of secretory vesicles at the cell surface. As no v SNARE has been shown to confer endocytosis, we examined whether yeast lacking the SNC genes, or possessing a temperature-sensitive allele of SNC1 (SNC1(ala43)), are deficient in the endocytic uptake of components from the cell surface. We found that both SNC and temperature-shifted SNC1(ala43) yeast are deficient in their ability to deliver the soluble dye FM4-64 to the vacuole. Under conditions in which vesicles accumulate, FM4-64 stained primarily the cytoplasm as well as fragmented vacuoles. In addition, alpha-factor-stimulated endocytosis of the alpha-factor receptor, Ste2, was fully blocked, as evidenced using a Ste2-green fluorescent protein fusion protein as well as metabolic labeling studies. This suggests a direct role for Snc v-SNAREs in the retrieval of membrane proteins from the cell surface. Moreover, this idea is supported by genetic and physical data that demonstrate functional interactions with t-SNAREs that confer endosomal transport (e.g., Tlg1,2). Notably, Snc1(ala43) was found to be nonfunctional in cells lacking Tlg1 or Tlg2. Thus, we propose that synaptobrevin/VAMP family members are engaged in anterograde and retrograde protein sorting steps between the Golgi and the plasma membrane. PMID- 11029061 TI - Wpkci, encoding an altered form of PKCI, is conserved widely on the avian W chromosome and expressed in early female embryos: implication of its role in female sex determination. AB - Two W chromosome-linked cDNA clones, p5fm2 and p5fm3, were obtained from a subtracted (female minus male) cDNA library prepared from a mixture of undifferentiated gonads and mesonephroi of male or female 5-d (stages 26-28) chicken embryos. These two clones were demonstrated to be derived from the mRNA encoding an altered form of PKC inhibitor/interacting protein (PKCI), and its gene was named Wpkci. The Wpkci gene reiterated approximately 40 times tandemly and located at the nonheterochromatic end of the chicken W chromosome. The W linkage and the moderate reiteration of Wpkci were conserved widely in Carinatae birds. The chicken PKCI gene, chPKCI, was shown to be a single-copy gene located near the centromere on the long arm of the Z chromosome. Deduced amino acid sequences of Wpkci and chPKCI showed approximately 65% identity. In the deduced sequence of Wpkci, the HIT motif, which is essential for PKCI function, was absent, but the alpha-helix region, which was conserved among the PKCI family, and a unique Leu- and Arg-rich region, were present. Transcripts from both Wpkci and chPKCI genes were present at significantly higher levels in 3- to 6-d (stages 20-29) embryos. These transcripts were detected in several embryonic tissues, including undifferentiated left and right gonads. When the green fluorescent protein-fused form of Wpkci was expressed in male chicken embryonic fibroblast, it was located almost exclusively in the nucleus. A model is presented suggesting that Wpkci may be involved in triggering the differentiation of ovary by interfering with PKCI function or by exhibiting its unique function in the nuclei of early female embryos. PMID- 11029063 TI - The phylogenetic utility of the codon-degeneracy model. AB - The codon-degeneracy model (CDM) predicts relative frequencies of substitution for any set of homologous protein-coding DNA sequences based on patterns of nucleotide degeneracy, codon composition, and the assumption of selective neutrality. However, at present, the CDM is reliant on outside estimates of transition bias. A new method by which the power of the CDM can be used to find a synonymous transition bias that is optimal for any given phylogenetic tree topology is presented. An example is illustrated that utilizes optimized transition biases to generate CDM GF-scores for every possible phylogenetic tree for pocket gophers of the genus Orthogeomys. The resulting distribution of CDM GF scores is compared and contrasted with the results of maximum parsimony and maximum likelihood methods. Although convergence on a single tree topology by the CDM and another method indicates greater support for that particular tree, the value of CDM GF-score as the sole optimality criterion for phylogeny reconstruction remains to be determined. It is clear, however, that the a priori estimation of an optimum transition bias from codon composition has a direct application to differentiating between alternative trees. PMID- 11029064 TI - Use of RNA secondary structure for studying the evolution of RNase P and RNase MRP. AB - Secondary structure is evaluated for determining evolutionary relationships between catalytic RNA molecules that are so distantly related they are scarcely alignable. The ribonucleoproteins RNase P (P) and RNase MRP (MRP) have been suggested to be evolutionarily related because of similarities in both function and secondary structure. However, their RNA sequences cannot be aligned with any confidence, and this leads to uncertainty in any trees inferred from sequences. We report several approaches to using secondary structures for inferring evolutionary trees and emphasize quantitative tests to demonstrate that evolutionary information can be recovered. For P and MRP, three hypotheses for the relatedness are considered. The first is that MRP is derived from P in early eukaryotes. The next is that MRP is derived from P from an early endosymbiont. The third is that both P and MRP evolved in the RNA-world (and the need for MRP has since been lost in prokaryotes). Quantitative comparisons of the pRNA and mrpRNA secondary structures have found that the possibility of an organellar origin of MRP is unlikely. In addition, comparison of secondary structures support the identity of an RNase P-like sequence in the maize chloroplast genome. Overall, it is concluded that RNA secondary structure is useful for evaluating evolutionary relatedness, even with sequences that cannot be aligned with confidence. PMID- 11029062 TI - Mutant RBL mast cells defective in Fc epsilon RI signaling and lipid raft biosynthesis are reconstituted by activated Rho-family GTPases. AB - Characterization of defects in a variant subline of RBL mast cells has revealed a biochemical event proximal to IgE receptor (Fc epsilon RI)-stimulated tyrosine phosphorylation that is required for multiple functional responses. This cell line, designated B6A4C1, is deficient in both Fc epsilon RI-mediated degranulation and biosynthesis of several lipid raft components. Agents that bypass receptor-mediated Ca(2+) influx stimulate strong degranulation responses in these variant cells. Cross-linking of IgE-Fc epsilon RI on these cells stimulates robust tyrosine phosphorylation but fails to mobilize a sustained Ca(2+) response. Fc epsilon RI-mediated inositol phosphate production is not detectable in these cells, and failure of adenosine receptors to mobilize Ca(2+) suggests a general deficiency in stimulated phospholipase C activity. Antigen stimulation of phospholipases A(2) and D is also defective. Infection of B6A4C1 cells with vaccinia virus constructs expressing constitutively active Rho family members Cdc42 and Rac restores antigen-stimulated degranulation, and active Cdc42 (but not active Rac) restores ganglioside and GPI expression. The results support the hypothesis that activation of Cdc42 and/or Rac is critical for Fc epsilon RI mediated signaling that leads to Ca(2+) mobilization and degranulation. Furthermore, they suggest that Cdc42 plays an important role in the biosynthesis and expression of certain components of lipid rafts. PMID- 11029065 TI - A new appraisal of the prokaryotic origin of eukaryotic phytochromes. AB - The evolutionary origin of the phytochromes of eukaryotes is controversial. Three cyanobacterial proteins have been described as "phytochrome-like" and have been suggested to be potential ancestors of these essential photoreceptors: Cph1 from Synechocystis PCC 6803, showing homology to phytochromes along its entire length and known to attach a chromophore; and PlpA from Synechocystis PCC 6803 and RcaE from Fremyella diplosiphon, both showing homology to phytochromes most strongly only in the C-terminal region and not known to bind a chromophore. We have reexamined the evolution of the photoreceptors using for PCR amplification a highly conserved region encoding the chromophore-binding domain in both Cph1 and phytochromes of plants and have identified genes for phytochrome-like proteins (PLP) in 11 very diverse cyanobacteria. The predicted gene products contain either a Cys, Arg, Ile, or Leu residue at the putative chromophore binding site. In 10 of the strains examined only a single gene was found, but in Calothrix PCC 7601 two genes (cphA and cphB) were identified. Phylogenetic analysis revealed that genes encoding PLP are homologues that share a common ancestor with the phytochromes of eukaryotes and diverged before the latter. In contrast, the putative sensory/regulatory proteins, including PlpA and RcaE, that lack a part of the chromophore lyase domain essential for chromophore attachment on the apophytochrome, are only distantly related to phytochromes. The Ppr protein of the anoxygenic photosynthetic bacterium Rhodospirillum centenum and the bacterial phytochrome-like proteins (BphP) of Deinococcus radiodurans and Pseudomonas aeruginosa fall within the cluster of cyanobacterial phytochromes. PMID- 11029067 TI - Measuring shifts in function and evolutionary opportunity using variability profiles: a case study of the globins. AB - Variability profiles measured over a set of aligned sequences can be used to estimate evolutionary freedom to vary. Differences in variability profiles between clades can be used to identify shifts in function at the molecular level. We demonstrate such a shift between the alpha and beta subunits of hemoglobin. We also show that the variability profiles for myoglobin are different between whales and primates and speculate that the differences between the two clades may reflect a shift associated with the novel oxygen storage demands in the lineage leading to whales. We discuss the relationship between sequence variability and "evolutionary opportunity" and explore the utility of Maynard Smith's multidimensional evolutionary opportunity space metaphor for exploring functional constraints, genetic redundancy, and the context dependency of the genotype phenotype map. This work has implications for quantitatively defining and comparing protein function. Supplementary data is available from bioinfo.mbb.yale. edu/align. PMID- 11029066 TI - MHC (major histocompatibility complex)-DRB genes and polymorphisms in common marmoset. AB - A New World monkey, the common marmoset (Callithrix jacchus), will be used as a preclinical animal model to study the feasibility of cell and gene therapy targeting immunological and hematological disorders. For elucidating the immunogenetic background of common marmoset to further studies, in the present study, polymorphisms of MHC-DRB genes in this species were examined. Twenty-one Caja-DRB exon 2 alleles, including seven new ones, were detected by means of subcloning and the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods followed by nucleotide sequencing. Based on the alignment of these allele sequences, we designed two pairs of specific primers and established a PCR-SSCP method for DNA-based histocompatibility typing of the common marmoset. According to the family segregation data and phylogenetic analyses, we presumed that Caja-DRB alleles could be classified into five different loci. Southern blotting analysis also supported the existence of multiple DRB loci. The patterns of nucleotide substitutions suggests that positive selection operates in the antigen-recognition sites of Caja-DRB genes. PMID- 11029068 TI - Fast evolution of interleukin-2 in mammals and positive selection in ruminants. AB - Interleukin-2 (IL-2) is a cytokine involved in induction and regulation of the immune response in mammals. There have been numerous reports about the search for IL-2 in species other than mammals, and recently an IL-2-like gene has been isolated in chicken. Using PCR, we searched for IL-2 gene sequences in a wide variety of mammals, including marsupials and monotremes, as well as in birds. Although we can readily amplify IL-2 gene fragments in placental mammals, no amplification was obtained in other species. This is best explained by very high substitution rates. This suggest that strategies to isolate IL-2 homologous genes outside mammals should involve functional assays, as for the chicken gene, and not hybridization-based techniques. Nonsynonymous substitution rates are especially high in ruminants, due to positive selection acting on regions important in term of structure-function. We suggest that, although globally similar, the immune response of various mammals is not identical, mainly at the level of cytokine-mediated regulations. PMID- 11029069 TI - Antigenic drift of viruses within a host: a finite site model with demographic stochasticity. AB - We theoretically study the antigenic drift of viruses within an infected host, as observed in human immunodeficiency virus (HIV) and equine infectious anemia virus (EIAV) infections, assuming that a finite number of antigen-determining sites at the viral envelop gene are responsible for the specific immune response. The pattern of antigen evolution becomes more complex than that predicted from the previous one-dimensional antigen space models. If the viral growth rate is sufficiently large, the demographic stochasticity for the fate of a new antigen mutant can be neglected. The high dimensionality in the way a virus escapes the immune defense in genotype space could then causes a rapid increase in the antigenic diversity and the total viral density, until finally the whole antigen genotypes are used up. The viral population is then driven to extinction in a host by the enhanced immune response to all genotypes. In contrast, if the viral growth rate is moderate or small so that only a small fraction of new antigen mutants can survive during the initial endangered period of random extinction, the viral antigenic diversity and the total density remain bounded, thereby enabling them to persist for a prolonged period by shifting the dominant antigen types. The phylogenetic pattern of antigen divergence is well characterized by the mean number of surviving antigen mutants from an antigen genotype. The substitution rate at antigen-determining sites increases as the efficiency of host immune response increases. PMID- 11029070 TI - Evolutionary history of B1 retroposons in the genus Mus. AB - Short interspersed DNA elements (SINEs) amplify by retroposition either by (i) successive waves of amplification from one or a few evolving master genes or by (ii) the generation of new master genes that coexist with their progenitors. Individual, highly conserved, elements of the B1 SINE family were identified from the GenBank nucleotide database using various B1 subfamily consensus query sequences to determine their integration times into the mouse genome. A comparison of orthologous loci in various species of the genus Mus demonstrated that four subfamilies of B1 elements have been amplifying within the last 1-3 million years. Therefore, B1 sequences are generated by coexisting source genes. Additionally, three B1 subfamilies have been concurrently propagated during subspecies divergence and strain formation in Mus, indicating very recent activity of this retroposon family. The patterns of intra- and interspecies variations of orthologous loci demonstrate the usefulness of B1 integrations as a phylogenetic tool. A single inconsistency in the phylogenetic trends was depicted by the presence of a B1 insert in an orthologous locus exclusively in M. musculus and M. pahari. However, DNA sequence analysis revealed that these were independent integrations at the same genomic site. One highly conserved B1 element that integrated at least 4-6 million years ago suggests the possibility of occasional function for B1 integrations. PMID- 11029071 TI - Evolutionary studies on uricases of fungal endosymbionts of aphids and planthoppers. AB - Aphids belonging to the three genera Tuberaphis, Glyphinaphis, and Cerataphis contain extracellular fungal symbionts that resemble endocellular yeast-like symbionts of planthoppers. Whereas the symbiont of planthoppers has a uricase (urate oxidase; EC 1.7.3.3) and recycles uric acid that the host stores, no uric acid was found in Tuberaphis styraci, and its fungal symbiont did not exhibit the uricase activity. However, the fungal symbionts of these aphids, including that of T. styraci, were shown to have putative uricase genes, or pseudogenes, for the uricase. Sequence analysis of these genes revealed that deleterious mutations occurred independently on each lineage of Glyphinaphis and Tuberaphis, while no such mutation was found in the lineage of Cerataphis. These genes were almost identical to those cloned from the symbionts of planthoppers, though the host aphids and planthoppers are phylogenetically distant. To estimate the phylogenetic relationship in detail between the fungal symbionts of aphids and those of planthoppers, a gene tree was constructed based on the sequences of the uricase genes including their flanking regions. As a result, the symbionts of planthoppers and Tuberaphis aphids formed a sister group against those of Glyphinaphis and Cerataphis aphids with high bootstrap confidence levels, which strongly suggests that symbionts have been horizontally transferred from the aphids' lineage to the planthoppers'. PMID- 11029072 TI - Ancyromonadida: a new phylogenetic lineage among the protozoa closely related to the common ancestor of metazoans, fungi, and choanoflagellates (Opisthokonta). AB - Molecular and morphological evidence points to the ancyromonad Ancyromonas as a plausible candidate for the closest relative to the common ancestor of metazoans, fungi, and choanoflagellates (the Opisthokonta). Using 18S rDNA sequences from most of the major eukaryotic lineages, maximum-likelihood, minimum-evolution, and maximum-parsimony analyses yielded congruent phylogenies supporting this hypothesis. Combined with ultrastructural similarities between Ancyromonas and opisthokonts, the evidence presented here suggests that Ancyromonas may form an independent lineage, the Ancyromonadida Cavalier-Smith 1997, closer in its relationship to the opisthokonts than is its nearest protist relatives, the Apusomonadida. However, the very low bootstrap support for deep nodes and hypothesis testing indicate that the resolving power of 18S rDNA sequences is limited for examining this aspect of eukaryotic phylogeny. Alternate branching positions for the Ancyromonas lineage cannot be robustly rejected, revealing the importance of ultrastructure when examining the origins of multicellularity. The future use of a multigene approach may additionally be needed to resolve this aspect of eukaryotic phylogeny. PMID- 11029073 TI - Histone H1 genes and histone gene clusters in the genus Drosophila. AB - Whereas the genomes of many organisms contain several nonallelic types of linker histone genes, one single histone H1 type is known in Drosophila melanogaster that occurs in about 100 copies per genome. Amplification of H1 gene sequences from genomic DNA of wild type strains of D. melanogaster from Oregon, Australia, and central Africa yielded numerous clones that all exhibited restriction patterns identical to each other and to those of the known H1 gene sequence. Nucleotide sequences encoding the evolutionarily variable domains of H1 were determined in two gene copies of strain Niamey from central Africa and were found to be identical to the known H1 sequence. Most likely therefore, the translated sequences of D. melanogaster H1 genes do not exhibit intragenomic or intergenomic variations. In contrast, three different histone H1 genes were isolated from D. virilis and found to encode proteins that differ remarkably from each other and from the H1 of D. melanogaster and D. hydei. About 40 copies of H1 genes are organized in the D. virilis genome with copies of core histone genes in gene quintets that were found to be located in band 25F of chromosome 2. Another type of histone gene cluster is present in about 15 copies per genome and contains a variable intergenic sequence instead of an H1 gene. The H1 heterogeneity in D. virilis may have arisen from higher recombination rates than occur near the H1 locus in D. melanogaster and might provide a basis for formation of different chromatin subtypes. PMID- 11029074 TI - What is characteristic of fungal lysine synthesis through the alpha-aminoadipate pathway? AB - Recent finding that a prokaryote synthesizes lysine through the alpha aminoadipate pathway demonstrates that the lysine synthesis through the alpha aminoadipate pathway is not typical of fungi. However, the fungal lysine biosynthesis is not completely the same as the prokaryotic one. We point out that alpha-aminoadipate reductase is a key enzyme to the evolution of fungal lysine synthesis. In addition, fungi have two different saccharopine dehydrogenases, which is also characteristic of fungi. PMID- 11029075 TI - Sequence, organization, and evolution of Rh50 glycoprotein genes in nonhuman primates PMID- 11029076 TI - Spatial Organization of Microbial Biofilm Communities. AB - The application of advanced microscopy and molecular and electrochemical high resolution methods has provided insights into the structural organization and function of biofilm communities. It appears that cellular properties such as growth differentiation, chemotaxis, and cell-to-cell signaling enable biofilm communities to organize structurally in response to the external conditions and the activities of the different biofilm members. Thereby resource utilization becomes optimized, and processes which require syntrophic relationships or special micro-environments become facilitated. PMID- 11029077 TI - Artificial Cyanobacterial Mats: Growth, Structure, and Vertical Zonation Patterns. AB - The formation of cyanobacterial mats (originally induced by incubation of sediment cores in which metazoans and most other eukaryotes had been removed) was followed over approximately 2.6 years. The thickness of the mats increased at a rate of 2-3 mm per year because of accumulation of empty cyanobacterial sheaths and as a result of carbonate deposition; the fraction of living biomass remained relatively constant over at least 2 years, but there was a slow accumulation of nonliving organic C ( approximately 1 mmol yr(-1)). Biota composition (dominated by five types of filamentous cyanobacteria, unicellular cyanobacteria, diatoms, anoxygenic phototrophs, and heterotrophic bacteria) and vertical zonation patterns in the upper 2-3 mm of the mats were also almost constant over time. Using transmission electron microscopy and stereological analysis it was possible to quantify the vertical distribution of major groups of organisms. PMID- 11029078 TI - Bio-optical Characteristics and the Vertical Distribution of Photosynthetic Pigments and Photosynthesis in an Artificial Cyanobacterial Mat. AB - Zonations of photosynthesis and photopigments in artificial cyanobacterial mats were studied with (i) oxygen and pH microsensors, (ii) fiber-optic microprobes for field radiance, scalar irradiance, and PSII fluorescence, and (iii) a light microscope equipped with a spectrometer for spectral absorbance and fluorescence measurements. Our analysis revealed the presence of several distinct 1-2 mm thick cyanobacterial layers mixed with patches of anoxygenic photosynthetic bacteria. Strong attenuation of visible light confined the euphotic zone to the uppermost 3 mm of the mat, where oxygen levels of 3-4 times air saturation and a pH peak of up to pH 8.8 were observed under saturating irradiance (413 umol photon m(-2) s( 1)). Oxygen penetration was 5 mm in light and decreased to 1 mm in darkness. Volumetric oxygen consumption in the photic and aphotic zones of illuminated mat was 5.5 and 2.9 times higher, respectively, than oxygen consumption in dark incubated mats. Scalar irradiance reached 100-150% of incident irradiance in the upper 0.5 mm of the mat due to intense scattering in the matrix of cells, exopolymers, and carbonate precipitates. In deeper mat layers scalar irradiance decreased nearly exponentially, and highest attenuation coefficients of 6-7 mm( 1) were found in cyanobacterial layers, where photosynthesis and photopigment fluorescence also peaked. Visible light was attenuated >100 times more strongly than near infrared light. Microscope spectrometry on thin sections of mats allowed detailed spectral absorbance and fluorescence measurements at defined positions relative to the mat surface. Besides strong spectral signals of cyanobacterial photopigments (Chl a and phycobiliproteins), the presence of both green and purple photosynthetic bacteria was evident from spectral signals of Bchl a and Bchl c. Microprofiles of photopigment absorbance correlated well with microdistributions of phototrophs determined in an accompanying study. PMID- 11029079 TI - Bacterioplankton Community Composition in Five Lakes Differing in Trophic Status and Humic Content. AB - To investigate the relation between lake type and bacterioplankton community composition, five Swedish lakes, which differed from each other in nutrient content and water color, were studied. Denaturing gradient gel electrophoresis (DGGE) of 16S rDNA was used to examine community composition. The DGGE-patterns of the different samples were analyzed in relation to physical, chemical, and biological data from the lakes by canonical correspondence analysis (CCA). The three variables found to most strongly correlate with the DGGE patterns were biomasses of microzooplankton, cryptophytes, and chrysophytes, suggesting that these biota had an impact on bacterioplankton community structure. Two of the three factors were, in turn, significantly correlated to parameters associated with the trophic status of the lakes, indicating that the nutrient content of the lakes, at least indirectly, influenced the structure of the bacterioplankton community. The relation to water color was less pronounced. PMID- 11029080 TI - Bacterial Growth Rate and Marine Virus-Host Dynamics. AB - The dynamics of a marine virus-host system were investigated at different steady state growth rates in chemostat cultures and the data were analyzed using a simple model. The virus-host interactions showed strong dependence on host cell growth rate. The duration of the infection cycle and the virus burst size were found to depend on bacterial growth rate, and the rate of cell lysis and virus production were positively correlated with steady state growth rate in the cultures (r(2) > 0.96, p < 0.05). At bacterial growth rates of 0.02 to 0.10 h(-1) in the chemostats the virus burst size increased from 12 +/- 4 to 56 +/- 4, and the latent period decreased from 2.0 to 1.7 h. Resistant clones of the host strain were present in the cultures from the beginning of the experiment and replaced the sensitive host cells following viral lysis in the cultures. Regrowth of resistant cells correlated significantly (r(2) = 1.000, p < 0.02) with the lysis rate of sensitive cells, indicating that release of viral lysates stimulated growth of the non-infected, resistant cells. The constructed model was suitable for simulating the observed dynamics of the sensitive host cells, viruses and resistant clones in the cultures. The model was therefore used in an attempt to predict the dynamics of this virus-host interaction in a natural marine environment during a certain set of growth conditions. The simulation indicated that a steady state relationship between the specific viruses and sensitive and resistant bacterial clones may occur at densities that are reasonable to assume for natural environments. The study demonstrates that basic characterization and modeling of specific virus-host interactions may improve our understanding of the behavior of bacteria and viruses in natural systems. PMID- 11029081 TI - Effects of Resources and Trophic Interactions on Freshwater Bacterioplankton Diversity. AB - In a study of bacterioplankton in an oligotrophic lake in northern Wisconsin, a community fingerprinting technique, automated ribosomal intergenic spacer analysis (ARISA), was used to determine the effect of resources and trophic interactions on bacterioplankton diversity. Inorganic nitrogen and phosphorus (NP), carbon in the form of glucose (G) or dissolved organic matter extracted from peat (DOM), and carbon and NP in combination were added to two types of experimental systems. Ten-liter mesocosms contained all components of the original aquatic community except for large zooplankton. One-liter dilution cultures were prepared so that the effects of grazers and phytoplankton were removed. During a 3-day incubation, bacterial production showed the greatest response to the carbon plus NP treatment in both experimental systems, but bacterial diversity was strikingly different between them. In the mesocosms, the number of ARISA-PCR fragments averaged 41 per profile, whereas the dilution culture communities were highly reduced in complexity, dominated in most cases by a single PCR fragment. Further analysis of the mesocosm data suggested that whereas the NPDOM addition caused the greatest aggregate bacterial growth response, the addition of NP alone caused the largest shifts in community composition. These results suggest that the measurement of aggregate responses, such as bacterial production, alone in studies of freshwater bacterial communities may mask the effects of resources on bacterioplankton. PMID- 11029082 TI - Improved Recoverability of Microbial Colonies from Marine Sponge Samples. AB - The growth of microorganisms from marine sponge samples was studied on various low to high nutrient solid media using media supplements. The supplements utilized were catalase, sodium pyruvate, and a combination of the two. Medium composition was found to influence the growth response on the supplemented media. Microorganisms on low nutrient media responded more favorably to the media additions than on high nutrient media. Thirty-five percent of the supplemented media demonstrated colony forming unit (CFU) recoveries that were 50% or greater than those of the unamended control plates. Twenty-one percent showed recoveries of more than 100% of the control values, with sodium pyruvate additions providing for the greatest overall increase in recovery, whether alone or in conjunction with catalase. These findings suggest that addition of catalase or sodium pyruvate to solid growth and isolation media may improve recoverability of microorganisms from natural samples. PMID- 11029083 TI - Relationships among Bacterial Cell Size, Productivity, and Genetic Diversity in Aquatic Environments using Cell Sorting and Flow Cytometry. AB - The study of relationships between cell size and productivity is of key importance in microbial ecology to understand which members of natural aquatic communities are responsible for the overall activity and/or productivity. Flow sorting of microorganisms from different environmental samples was used to analyze the activity of bacterial cells depending on their biovolume. Bacterial cells from five different natural samples taken along the Mediterranean coast including fresh- and seawaters were incubated with tritiated leucine, then stained with SYTO 13 and sorted by flow cytometry according to their average side angle-scattered (SSC) light. In all samples, a bell-shaped relationship was found between cell biovolume and activity, whereas activity of a given cell-size class varied between samples. In contrast, an inverse relationship was found between biovolumes and abundances. These results suggest that medium-sized cells with highest growth rates are probably submitted to intense grazing. For one sample, bacteria within five different size classes were sorted and the genetic diversity of cells within each sorted size class and that of the whole community were analyzed by the denaturing gradient gel electrophoresis (DGGE) method. The genetic diversity, as determined at the community level was highly represented into the pool of small cells, whereas only few species were present into larger cell subpopulations. The results suggest that only a few genotypes may be dominant within the largest and most productive cells. Furthermore, cell size polymorphism as well as heterogeneous cellular activities were found within some species. PMID- 11029084 TI - Response of Antarctic Soil Bacterial Assemblages to Contamination by Diesel Fuel and Crude Oil. AB - The effects of diesel fuel and "Arabian light" crude oil addition on Antarctic bacterial assemblages were studied in four contaminated soils during 1 year in the Terre Adelie land area. Monthly sampling allowed a regular survey of the bacterial changes occurring in the contaminated soils. All samples were analyzed for total bacteria, heterotrophic culturable microbiota, and hydrocarbon utilizing microbiota. Crude oil contamination induced an initial increase of all bacterial parameters in all contaminated soils. Diesel oil contamination had a more complex effect. Hydrocarbon degrading bacterial abundance increases occurred after diesel oil addition. In contrast, general heterotrophic bacterial abundance could significantly decrease in the same conditions. In all cases the stimulatory effects of oil addition disappeared after several months of contamination. PMID- 11029085 TI - Transjugular retrograde obliteration for chronic portosystemic encephalopathy. AB - Chronic portosystemic encephalopathy (CPSE) is uncommon, and its management has yet to be determined. We have been able to control five cases of CPSE using transjugular retrograde obliteration (TJO), and we report our clinical results with this technique. All of the five patients were suffering from cirrhosis and had gastric varices and large gastrorenal shunts. According to Sherlock's classification, the grade of encephalopathy was II in two patients, III in two, and IV in one. According to Child's classification, one had class B and four had class C cirrhosis. TJO was performed using a 6-F angiographic catheter with an occlusive balloon 20 mm in diameter. Absolute ethanol and 5% ethanolamine oleate with iopamidol were used to obliterate the gastrorenal shunt. The gastrorenal shunt was successfully obliterated, and the encephalopathy improved to grade 0 after TJO in all cases. The portal flow volume increased significantly from 542 +/- 189 to 992 +/- 139 mL/min (p < 0.01). The plasma ammonia levels before and after TJO were 189 +/- 40 and 51 +/- 23 microg/dL, and the indocyanine green retention rates at 15 min were 44 +/- 13% and 27 +/- 12%, with both changes being significant (p < 0.01). Minor complications observed were fever of over 38 degrees C and tarry stools due to hemorrhagic gastritis in one patient, which was being controlled conservatively. One patient died of hepatocellular carcinoma 27 months after TJO. The other four patients survived without recurrence of CPSE 17 74 months (44 +/- 24 months) after TJO. We conclude that TJO can be adopted as a safe and effective treatment for CPSE. PMID- 11029086 TI - Congenital extrahepatic portocaval shunt associated with hepatic hyperplastic nodules in a patient with Dubin-Johnson syndrome. AB - We report a rare case of congenital extrahepatic portocaval shunt diagnosed during evaluation of hyperplastic nodules in the liver. Diagnostic imagings showed hypoplasia of the intrahepatic portal venous system and splanchnic portal venous return to the inferior vena cava through aberrant vessels. Altered hepatic blood flow dynamics due to this shunt may have been implicated in the etiology of the hepatic hyperplastic nodules. PMID- 11029087 TI - CO(2) wedged hepatic venography: technical considerations and comparison with direct and indirect portography with iodinated contrast. AB - BACKGROUND: We evaluated the efficacy and safety of CO(2) wedged hepatic venography (CO(2) WHV) by comparing it with direct transjugular (DP) and indirect arterial portography (IP). METHODS: Twenty-one CO(2) WHV and IP examinations were performed in 20 patients; 13 of them also underwent DP within 48 h of CO(2) WHV and IP. IP involved the injection of iodinated contrast into the superior mesenteric and splenic arteries. DP was performed from a transjugular approach, during transjugular intrahepatic portosystem shunt placement, with the injection of iodinated contrast into the superior mesenteric or splenic vein. The parameters evaluated were visualization of vessels and varices, portal vein thrombosis detection, and complications. RESULTS: CO(2) WHV depicted the splenic vein in 57%, the superior mesenteric vein in 62%, the main portal vein in 90%, the right portal vein in 95%, and the left portal vein in 90% of patients. It also demonstrated gastroesophageal varices in seven cases, a splenorenal shunt in one case, mesenteric varices in one case, and a recanalized umbilical vein in one case; other varices were also seen. CONCLUSION: CO(2) WHV is a good and safe technique for demonstrating the portal circulation. It may provide information not obtainable by IP and DP. However, IP provides better demonstration of the variceal network. PMID- 11029088 TI - Computed tomography during arterial portography under temporary balloon occlusion of the hepatic artery: evaluation of pseudolesions caused by arterio-portal venous shunts. AB - BACKGROUND: Computed tomography during arterial portography (CTAP) under temporary balloon occlusion of the hepatic artery (BOHA-CTAP) was introduced to evaluate pseudolesions caused by portal venous impairments such as arterioportal shunt and tumor thrombus. METHODS: BOHA-CTAP was performed in seven patients with hepatocellular carcinoma and correlated with clinical outcomes. For patients with wedge-shaped defects suggestive of pseudolesions, BOHA-CTAP was obtained by a 5-F balloon occlusion catheter into the proper hepatic artery through the second 5-F introducer inserted into the common femoral artery a few centimeters below the first 5-F sheath for CTAP. RESULTS: Eight pseudolesions were determined clinically on follow-up CT, ultrasonography, or magnetic resonance imagings. On BOHA-CTAP, five of the eight pseudolesions were eliminated, and two were diminished in comparision with conventional CTAP. One wedge-shaped defect due to tumor thrombus in the portal vein did not show any change. CONCLUSION: BOHA-CTAP can reduce pseudolesions caused by portal venous impairments and enable the demarcation of the true tumors. PMID- 11029089 TI - Catheter dislodgement of percutaneous transhepatic biliary drainage: identification of role of puncture sites and catheter sheath. AB - BACKGROUND: To identify the appropriate puncture points in the bile duct to avoid catheter dislodgement. METHODS: Percutaneous transhepatic biliary drainage catheters (n = 300) were placed in 242 patients. The frequency of dislodgement (complete dislodgement or bending of the catheter) was prospectively investigated. The puncture site of the bile duct was classified on the ultrasonographic findings as follows: Main-B3, main branch of the lateral inferior segment; peripheral-B3, peripheral branch of the lateral inferior segment; B2, lateral superior segment; left hepatic duct, proximal portion of the left hepatic duct; B8, anterior superior segment; B5, anterior inferior segment; B5 + 8, main bile duct of the anterior segment; B6, bile duct of posterior inferior segment; and right hepatic duct, proximal portion of the right hepatic duct. RESULTS: When a catheter without an outer sheath was used, catheter dislodgement in peripheral-B3 (2/11, 18%) was more common than in main-B3 (0/32, 0%; p < 0.05). In B5, catheter dislodgement (6/12, 50%) was more frequent than in B8 (3/20, 15%; p < 0.05) and in B6 (0/14, 0%; p < 0.005). When a catheter with an outer sheath was used, catheter dislodgement (2/207, 1%) was rare. CONCLUSION: Drainage from B5 and peripheral-B3 is associated with a high risk of dislodgement of the catheter. A catheter with an outer sheath was useful to prevent catheter dislodgement. PMID- 11029090 TI - Macronodular hepatic deformity on normal liver. AB - BACKGROUND AND METHODS: Macronodular hepatic deformity on normal liver is very rare. We present nine such cases and try to define the characteristic clinical and ultrasound (US) findings. RESULTS: In the left lobe, the lateral segment was replaced by multiple nodules and the medial segment was very atrophied and irregularly shaped. Compared with the left lobe, the right lobe showed very few abnormalities except for segment 6, which showed a macronodular deformity. These nodules, regardless of diameter or lobe, showed a relatively homogeneous internal structure and were isoechoic relative to the surrounding hepatic parenchyma. These multinodular changes on US corresponded to multiple regenerative nodules identified at laparoscopic evaluation. On color Doppler US and angiography, the major intrahepatic vessels were patent in all cases. CONCLUSION: Although relatively rare, the macronodular hepatic deformity on normal liver collected in our series showed a characteristic appearance by US. A good understanding of its characteristics may help sonographers in differentiating it from other, more common hepatic deformities. PMID- 11029091 TI - Liver tumors in children and young patients: sonographic and color Doppler findings. AB - BACKGROUND: Liver tumors are a relatively rare pathologic condition in children and young patients. The aim of the present study was to categorize the sonographic (US) and color Doppler results of liver tumors in these patients. METHODS: We retrospectively reviewed the US findings of 23 such cases: malignant tumor (13 cases)-hepatoblastoma (four cases), hepatocellular carcinoma (HCC; four cases), and hepatic metastasis (five cases); benign tumor (10 cases) hepatocellular adenoma (four cases), focal nodular hyperplasia (two cases), mesenchymal hamartoma (two cases), cystadenoma (one case), and hemangioendothelioma (one case). RESULTS: There was no specific US findings for each tumor type. HCC usually developed on a normal liver and was imaged as multiple nodules. Color Doppler US helped in differentiating multiple metastatic nodules (hypovascular) from multiple HCC nodules (hypervascular). Presence of intratumoral cystic areas was usually suggestive of benign tumors. Follow-up US was useful for detecting small nodules in high-risk groups (congenital biliary atresia, glycogen storage disease). Color Doppler US helped in diagnosing portal thrombus or intratumoral shunt. CONCLUSION: Although there were no highly specific findings, US and color Doppler results contributed, to a certain degree, to the diagnosis of liver tumors in children and young adults by showing intratumoral cystic areas or vascularity. PMID- 11029092 TI - Neoplasms containing normal hepatic vessels: imaging features. AB - BACKGROUND: We investigated the incidence and imaging features of hepatic neoplasms containing normal hepatic vessels. METHODS: Among 3183 patients with various hepatic neoplasms, we found nine patients with normal hepatic vessels traversing hepatic neoplasms. The presence of mass effect on the vessel traversing hepatic neoplasms was evaluated. Other suggestive findings of neoplasms such as altered hepatic contour, portal vein thrombosis, mass effect on extratumoral vessel, and bile duct dilatation were analyzed. RESULTS: Thirteen hepatic vessels (nine hepatic veins and four portal veins) extended through hepatic neoplasms in nine patients. Undisturbed hepatic vessels within the neoplasms were found in five patients with either primary or metastatic hepatic neoplasm. In one patient with undisturbed hepatic vessels within the neoplasm, there were no associated abnormal findings such as biliary dilatation, change of hepatic contour, or any changes involving the vessels external to neoplasms. CONCLUSION: Although rare, various primary and secondary hepatic neoplasms can have normal hepatic vessels passing through them without mass effect. However, correct diagnosis in most neoplasms would be possible with careful examination of associated findings. PMID- 11029093 TI - Multiphase helical CT findings after percutaneous ablation procedures for hepatocellular carcinoma. AB - BACKGROUND: Multiple-phase helical computed tomography (CT) has been regarded as the method of choice in the evaluation of patients with hepatocellular carcinoma (HCC) treated by nonsurgical procedures. The aim of this article was to report our experience in the assessment of nodular and parenchymal changes recognizable after various percutaneous ablation therapies. METHODS: We reviewed the studies of 116 consecutive patients with HCC treated with multisession percutaneous ethanol injection (56 patients, 98 nodules), single-session percutaneous ethanol injection (14 patients, 31 nodules), radiofrequency thermal ablation (32 patients, 48 nodules), and interstitial laser photocoagulation (14 patients, 25 nodules). CT had been performed 3-28 days after the last session (mean = 18 days) with unenhanced helical acquisition and with contrast-enhanced double- or triple phase helical acquisition. RESULTS: Persisting neoplastic tissue was identified within 54.5% of the nodules. It was located centrally in 4.5% of these nodules, peripherally in 11%, and eccentrically in 84.5%, and its shape was crescent in 58%, globular in 24.5%, and other in 16%. On arterial phase scans, viable tumor was hyperdense in 97% of the lesions and isodense in 3%; on portal phase scans, the tumor was hyperdense in 20%, isodense in 28%, and hypodense in 52%; on delayed phase scans, the tumor was consistently hypodense. Tumor necrosis was always hypodense on contrast-enhanced scans. On unenhanced images, 7.4% of the nodules were undetectable. Nodule diameter appeared as unchanged in 53% of the nodules and as larger in 47%; its shape was unchanged in 54% and modified in 46%; its margins were unchanged in 36% and modified in 64%. A rim of granulation tissue was detected around 15% of the nodules, and a perilesional transient attenuation difference was detected in 21%. Perihepatic effusion was seen in 13% of the patients, segmental biliary duct dilation and local atrophy each in 9%, arterioportal fistula in 6%, portal vein thrombosis, subcapsular collection and pleural effusion each in 7%, hepatic infarction in 5%, and inferior vena cava thrombosis in 2%. CONCLUSION: Percutaneous ablation of HCC may cause several changes. Knowledge of their CT appearance is mandatory to correctly assess and manage this tumor. PMID- 11029095 TI - Radiologic diagnosis of common bile duct stones. AB - Unenhanced spiral CT and, in the hands of the experienced, US are accurate noninvasive methods of imaging the common bile duct. This is particularly true when there is a low clinical and laboratory probability of a common bile duct stone being present. In the diagnosis of bile duct stones, CT and US are effective and readily available screening techniques and may aid in the selection of patients who require MRCP or treatment with ERCP. Avoiding ERCP and the associated expense and morbidity is a goal that is becoming attainable with optimized CT, US, and MRCP techniques and an awareness of subtle findings. PMID- 11029094 TI - Peripheral cholangiocarcinoma with extensive tumor thrombi in the portal venous system: case report. AB - Peripheral cholangiocarcinoma is a primary liver tumor, with some features that distinguish it from hepatocellular carcinoma, especially the low incidence of vascular involvement relative to mass size. Radiologically, tumor thrombus seen as an expanding central filling defect is a rare finding of cholangiocarcinoma on computed tomography, even though microscopic invasion or vascular encasement has been reported in a few studies. We report a case of a peripheral cholangiocarcinoma with tumor thrombus expanding and spreading from the portal vein to the superior mesenteric vein. PMID- 11029096 TI - Sonographic diagnosis of congenital tuberculosis: an experience with four cases. AB - Abdominal sonography of four infants with pyrexia and hepatomegaly demonstrated multiple hypoechoeic hepatic and splenic foci, guided biopsies of which showed caseating granulomas with acid-fast bacilli. Evidence of tuberculosis in maternal endometrium and its exclusion in the contacts further confirmed a diagnosis of congenital tuberculosis. Clinical suspicion supplemented by careful sonography facilitated early detection and antemortem diagnosis of this potentially fatal disease. PMID- 11029097 TI - Abdominal tuberculous lymphadenopathy: MR imaging findings. AB - BACKGROUND: The purpose of this study was to evaluate the utility of magnetic resonance imaging (MRI) as a diagnostic tool in abdominal tuberculous lymphadenopathy. METHODS: MRI studies of 11 patients with histologically proven abdominal tuberculous lymphadenopathy were reviewed with regard to anatomic distribution, size, shape, degree, and pattern of enhancement and relation of the lesions to adjacent structures. RESULTS: The most common site of involvement was the periportal area (n = 6), followed by the peripancreatic (n = 5), mesenteric (n = 1), and paraaortic (n = 1) areas. Eight patients were readily diagnosed as having tuberculous lymphadenopathy on abdominal computed tomography. Three patients had a heterogeneously enhancing masslike lesion adjacent to the pancreas and were initially diagnosed as having cystic tumor of the pancreas. On MRI, 11 lesions showed T1 iso- or hypointensity and central T2 hyperintensity. Two lesions showed T1 iso- or hypointensity and central T2 hypointensity. The lesions with different T2 signal intensities showed different patterns of enhancement on contrast-enhanced dynamic studies. The relations between the enlarged lymph nodes and adjacent bile ducts or vascular structures were well depicted on MRI. CONCLUSION: MRI was useful in differentiating enlarged lymph nodes abutting the pancreas initially diagnosed as cystic neoplasms on abdominal computed tomography. PMID- 11029098 TI - CT fluoroscopic guidance for percutaneous needle placement into abdominopelvic lesions with difficult access routes. AB - BACKGROUND: We wished to evaluate the utility of computed tomography fluoroscopy (CTF) for guiding percutaneous abdominopelvic biopsies or fluid aspirations that are considered difficult with conventional computed tomographic (CT) guidance. METHODS: CTF-guided percutaneous biopsy (n = 11) or fluid aspiration (n = 2) was attempted in 13 patients with lesions that were otherwise difficult or potentially unsafe by conventional CT guidance because they were deep to colon, small intestine, or major blood vessels. RESULTS: Using CTF assistance to guide external compression or needle positioning, appropriate needle placement was performed in 11 patients. Biopsy or aspiration was diagnostic in 10 patients. Needle advancement was not attempted in two patients. CONCLUSION: CTF appears to be a valuable tool to dynamically assist percutaneous needle placement into lesions that may be considered difficult with standard CT assistance. PMID- 11029099 TI - Simple and effective technique of guided biopsy in a closed MRI system. AB - BACKGROUND: Magnetic resonance (MR)-guided biopsies are generally regarded as complex interventions. We implemented interventional MR (IMR) with the resources available in and for practical application in a large (1100 beds) central hospital. METHODS: This simple and straightforward IMR technique uses a step-by step approach for localization, access route planning, biopsy, verification in at least two planes, and postoperative control. The technique has been used and evaluated unchanged for more than 400 punctures and interventions. RESULTS: Contrast, signal, matrix options, and visibility of needle track and tip permit uncomplicated orientation. The mean duration of a biopsy is 19 min. The technique can be applied to all radiologic puncture settings without any technical or medical complications. CONCLUSION: The crucial step in implementing IMR is not to contemplate its application but to simply start applying the procedure. PMID- 11029100 TI - Logistic advantages of four-section helical CT in the abdomen and pelvis. AB - BACKGROUND: Multisection helical computed tomography (CT) has the potential for providing data sets with better section profiles, more anatomic coverage, and shorter breath-holding periods. Our purpose was to quantitate these advantages in a clinical setting when imaging the abdomen and pelvis. METHODS: CT parameters including collimation, timing, z-axis coverage, and milliamperes were gathered retrospectively for the image set of both single-section (GE CT/i with 0.8-s rotation) and four-section (GE QX/i Lightspeed with 0.8-s rotation) helical CT scanners. Data were recorded for the abdomen and pelvis CT (n = 30 each), dual phase liver CT including the pelvis (n = 15 each), and dual-phase pancreas CT (n = 15 each). RESULTS: The abdominal and pelvic CT averaged 128.4 +/- 5.4 s for single-section scanners (70-s delay, two breath-holds of 21.1 and 17. 7 s with a 19.5-s interscan delay) and 92.2 +/- 2.2 s for the four-section scanner (70-s delay and a 22.2-s breath-hold; p < 0. 0001). For the dual liver and pelvis CT, single-section scanners averaged 119.9 +/- 7.5 s (30-s delay, 15.8-s arterial phase, 20.0-s interscan delay, 21.2-s venous phase, 19.5-s interscan delay, and 14. 2 s for the remaining abdomen and pelvis), whereas the four-section scanner averaged 86.8 +/- 2.5 s (30-s delay, 6.7-s arterial phase, 27.9-s interscan delay, and 21.8-s venous phase including the pelvis; p < 0.0001). For the dual pancreas CT, single-section scanners averaged 86.7 +/- 2.5 s (20-s delay, 28.3-s arterial phase, 17.8-s interscan delay, 21.7-s venous phase), whereas the four section scanner averaged 78.0 +/- 2.9 s (20-s delay, 9.7-s arterial phase, 30.7-s interscan delay, 13.0-s venous phase; p < 0. 0001). CONCLUSION: CT scanners having four-section technology can reduce overall data acquisition times by 10 30% and total milliamperes by 50-60% depending on the protocol with thinner slice profiles. PMID- 11029101 TI - Renal angiomyolipoma: evolutional changes of its internal structure on CT. AB - BACKGROUND: We evaluated interval changes of the internal structure of renal angiomyolipomas in relation to the doubling time on computed tomography (CT). METHODS: A retrospective review of 42 renal angiomyolipomas in 10 patients who had been followed up for more than 5 months with CT was performed. The doubling time of each tumor was estimated. We classified the internal structures of the tumors into five types, based on the fat component percentage. We compared changes between initial and follow-up CT, and the relationship with the doubling time was evaluated. RESULTS: The fat component increased more than the soft tissue component in 15 tumors (36%). The soft tissue component increased more than the fat component in only one tumor, in which an intratumoral hemorrhage occurred during the follow-up period. Twenty-six tumors (62%) showed no change. Most tumors with relatively long doubling times did not show a change in their internal structures. CONCLUSION: The growth of renal angiomyolipoma is due mainly to an increase in its fat component. An increase in the soft tissue component suggests the development of an intratumoral hemorrhage. PMID- 11029102 TI - Uterine lipoleiomyomas: US and CT findings. AB - Fatty tumors of the uterus are rare. The reported incidence differs, from 0.03% to 0.2%. Two new cases of uterine lipoleiomyoma are presented with ultrasound, computed tomography, and pathologic correlation. To date, only seven cases have been reported in the radiologic literature. PMID- 11029103 TI - Injected periurethral collagen for postprostatectomy urinary incontinence: MR and CT appearance. AB - BACKGROUND: Urinary incontinence, a disturbing complication of radical prostatectomy, is often treated with periurethral collagen injections to increase urethral closure and resistance to urinary outflow. METHODS: Using magnetic resonance imaging and computed tomography, we studied the appearance of glutaraldehyde cross-lined bovine collagen endoscopically injected into the periurethral tissues in four men who developed urinary incontinence after radical prostatectomy. Collagen was also scanned in vitro to verify its magnetic resonance appearance. RESULTS: Collagen deposits appear as well-circumscribed nodules of low to intermediate signal intensity on both T1- and T2-weighted images in the periurethral tissues or in the base of the subjacent penile bulb (base of corpus spongiosum). On contrast-enhanced computed tomography, collagen appears as a hypoattenuating nodular-filling defect within the penile bulb. CONCLUSION: These imaging characteristics should allow differentiation of collagen from locally recurrent prostate carcinoma and avoid inappropriate work up of benign findings. PMID- 11029109 TI - A tribute to our reviewers PMID- 11029104 TI - Arterial supply of liver segment IV. PMID- 11029110 TI - Choosing to encourage or discourage: perceived effectiveness of prescriptive versus proscriptive messages. AB - The estimated cost of repairing damage caused to recreational sites annually is in the hundreds of millions of dollars. These depreciative activities also reduce the quality of visitors' experiences in the damaged areas. Indirect methods, such as visitor education through brochures and signs, continue to be the least controversial management approaches to depreciative acts. Yet, the literature on studies examining the most effective message presentations remains sparse. A survey mailed to randomly selected National Association for Interpretation members assessed the perceived effectiveness of communications that encouraged positive conduct (prescriptive messages) versus those that discouraged negative conduct (proscriptive messages) in wildland and urban settings. Almost invariably, respondents viewed the encouragement-based prescriptive messages as more effective than the discouragement-based proscriptive messages. This finding stands in sharp contrast to an earlier study that discovered a preponderance of proscriptive versus prescriptive messages on signs in both wildland and urban recreational environments. Thus, although the great majority of interpreters see the encouragement of positive conduct as more effective, in practice, messages on signs are much more likely to discourage negative conduct. Reasons for this discrepancy are considered. PMID- 11029111 TI - Using multicriteria methods in environmental planning and management. AB - In environmental planning and decision processes several alternatives are analyzed in terms of multiple noncommensurate criteria, and many different stakeholders with conflicting preferences are involved. Based on our experience in real-life applications, we discuss how multicriteria decision aid (MCDA) methods can be used successfully in such processes. MCDA methods support these processes by providing a framework for collecting, storing, and processing all relevant information, thus making the decision process traceable and transparent. It is therefore possible to understand and explain why, under several conflicting preferences, a particular decision was made. The MCDA framework also makes the requirements for new information explicit, thus supporting the allocation of resources for the process. PMID- 11029112 TI - Water quality in drinking water reservoirs of a megacity, istanbul. AB - Providing clean water at relevant quality and quantity is a challenge that regulatory authorities have to face in metropolitan cities that seem to develop at their limits of sustainability. Istanbul strives to face such a challenge for its population of over 10 million, through six surface water resources. Two approaches of classification for the reservoirs are presented, one based on current regulations and an alternative based on a more detailed classification. The results have shown that nutrient control is the primary issue, and one of the reservoirs has already exceeded the limits of being eutrophic, one is at mesotrophic conditions, and the remaining four are at the limit of being eutrophic, indicating the significance of making the correct decision and taking pertinent measures for management and control. It has been observed that the only mesotrophic resource, which also has the best general quality class, has no industry and a very low population density, whereas the one that is already eutrophic is also the one with the lowest quality class, has the highest population density, and has the greatest percentage of urban land use within its watershed. PMID- 11029113 TI - Benefit-Cost Analysis of StormwaterQuality Improvements. AB - The major purpose of this paper is to explore the potential value of benefit-cost evaluation for stormwater quality management decisions at a local level. A preliminary benefit-cost analysis (BCA) screening method is used for maximum extent practicable (MEP) analysis, identifying promising management practices, and identifying societal and economic tradeoffs for local stormwater problems. Ballona Creek, a major urban storm drain in Los Angeles, California, USA, is used to illustrate the practicality of the benefit-cost evaluation. The Ballona Creek example demonstrates the economic limits of stormwater management in an urban region and attests to the value of coordinated basinwide management compared to uncoordinated management by individual landowners. Evaluation results suggest that in urban areas, the benefit of stormwater quality improvements might be far greater if accompanied by comprehensive redesign of drainage networks and neighboring land uses. In this case, benefit-cost analysis is found to be useful for evaluating and understanding stormwater management alternatives despite the uncertainties in characterizing stormwater quality and the effects of stormwater management on improving receiving water quality. PMID- 11029114 TI - Impacts of rotational grazing and riparian buffers on physicochemical and biological characteristicsof southeastern Minnesota, USA, streams. AB - We assessed the relationship between riparian management and stream quality along five southeastern Minnesota streams in 1995 and 1996. Specifically, we examined the effect of rotationally and continuously grazed pastures and different types of riparian buffer strips on water chemistry, physical habitat, benthic macroinvertebrates, and fish as indicators of stream quality. We collected data at 17 sites under different combinations of grazing and riparian management, using a longitudinal design on three streams and a paired watershed design on two others. Continuous and rotational grazing were compared along one longitudinal study stream and at the paired watershed. Riparian buffer management, fenced trees (wood buffer), fenced grass, and unfenced rotationally grazed areas were the focus along the two remaining longitudinal streams. Principal components analysis (PCA) of water chemistry, physical habitat, and biotic data indicated a local management effect. The ordinations separated continuous grazing from sites with rotational grazing and sites with wood buffers from those with grass buffers or rotationally grazed areas. Fecal coliform and turbidity were consistently higher at continuously grazed than rotationally grazed sites. Percent fines in the streambed were significantly higher at sites with wood buffers than grass and rotationally grazed areas, and canopy cover was similar at sites with wood and grass buffers. Benthic macroinvertebrate metrics were significant but were not consistent across grazing and riparian buffer management types. Fish density and abundance were related to riparian buffer type, rather than grazing practices. Our study has potentially important implications for stream restoration programs in the midwestern United States. Our comparisons suggest further consideration and study of a combination of grass and wood riparian buffer strips as midwestern stream management options, rather than universally installing wood buffers in every instance. RID="" ID="" The Unit is jointly sponsored by the US Geological Survey, Biological Resources Division; the Minnesota Department of Natural Resources; the University of Minnesota; and the Wildlife Management Institute. PMID- 11029115 TI - Assessing Watershed-Scale, Long-Term Hydrologic Impacts of Land-Use Change Using a GIS-NPS Model. AB - Land-use change, dominated by an increase in urban/impervious areas, has a significant impact on water resources. This includes impacts on nonpoint source (NPS) pollution, which is the leading cause of degraded water quality in the United States. Traditional hydrologic models focus on estimating peak discharges and NPS pollution from high-magnitude, episodic storms and successfully address short-term, local-scale surface water management issues. However, runoff from small, low-frequency storms dominates long-term hydrologic impacts, and existing hydrologic models are usually of limited use in assessing the long-term impacts of land-use change. A long-term hydrologic impact assessment (L-THIA) model has been developed using the curve number (CN) method. Long-term climatic records are used in combination with soils and land-use information to calculate average annual runoff and NPS pollution at a watershed scale. The model is linked to a geographic information system (GIS) for convenient generation and management of model input and output data, and advanced visualization of model results.The L THIA/NPS GIS model was applied to the Little Eagle Creek (LEC) watershed near Indianapolis, Indiana, USA. Historical land-use scenarios for 1973, 1984, and 1991 were analyzed to track land-use change in the watershed and to assess impacts on annual average runoff and NPS pollution from the watershed and its five subbasins. For the entire watershed between 1973 and 1991, an 18% increase in urban or impervious areas resulted in an estimated 80% increase in annual average runoff volume and estimated increases of more than 50% in annual average loads for lead, copper, and zinc. Estimated nutrient (nitrogen and phosphorus) loads decreased by 15% mainly because of loss of agricultural areas. The L THIA/NPS GIS model is a powerful tool for identifying environmentally sensitive areas in terms of NPS pollution potential and for evaluating alternative land use scenarios for NPS pollution management. PMID- 11029116 TI - Development of a methodology for selecting criteria and indicators of sustainable forest management: a case study on participatory assessment. AB - This paper describes an application of multiple criteria analysis (MCA) in assessing criteria and indicators adapted for a particular forest management unit. The methods include: ranking, rating, and pairwise comparisons. These methods were used in a participatory decision-making environment where a team representing various stakeholders and professionals used their expert opinions and judgements in assessing different criteria and indicators (C&I) on the one hand, and how suitable and applicable they are to a forest management unit on the other. A forest concession located in Kalimantan, Indonesia, was used as the site for the case study. Results from the study show that the multicriteria methods are effective tools that can be used as structured decision aids to evaluate, prioritize, and select sets of C&I for a particular forest management unit. Ranking and rating approaches can be used as a screening tool to develop an initial list of C&I. Pairwise comparison, on the other hand, can be used as a finer filter to further reduce the list. In addition to using these three MCA methods, the study also examines two commonly used group decision-making techniques, the Delphi method and the nominal group technique. Feedback received from the participants indicates that the methods are transparent, easy to implement, and provide a convenient environment for participatory decision making. PMID- 11029117 TI - Lightning fires in a brazilian savanna national park: rethinking management strategies. AB - Fire occurrences and their sources were monitored in Emas National Park, Brazil (17 degrees 49'-18 degrees 28'S; 52 degrees 39'-53 degrees 10'W) from June 1995 to May 1999. The extent of burned area and weather conditions were registered. Forty-five fires were recorded and mapped on a GIS during this study. Four fires occurred in the dry winter season (June-August; 7,942 ha burned), all caused by humans; 10 fires occurred in the seasonally transitional months (May and September) (33,386 ha burned); 31 fires occurred in the wet season, of which 30 were caused by lightning inside the park (29,326 ha burned), and one started outside the park (866 ha burned). Wet season lightning fires started in the open vegetation (wet field or grassy savanna) at a flat plateau, an area that showed significantly higher fire incidence. On average, winter fires burned larger areas and spread more quickly, compared to lightning fires, and fire suppression was necessary to extinguish them. Most lightning fires were patchy and extinguished primarily by rain. Lightning fires in the wet season, previously considered unimportant episodes, were shown to be very frequent and probably represent the natural fire pattern in the region. Lightning fires should be regarded as ecologically beneficial, as they create natural barriers to the spread of winter fires. The present fire management in the park is based on the burning of preventive firebreaks in the dry season and exclusion of any other fire. This policy does not take advantage of the beneficial effects of the natural fire regime and may in fact reduce biodiversity. The results presented here stress the need for reevaluating present policies and management procedures concerning fire in cerrado conservation areas. PMID- 11029118 TI - Monitoring the response of butterfly communities to prescribed fire. AB - Federal land managers in the western United States are interested in the potential of prescribed fire as a tool to decrease fuel loads, increase vegetational heterogeneity, and increase faunal diversity in various ecosystems. I tested whether implementation of a prescribed fire program by the US Forest Service in a watershed in the central Great Basin had significant effects on butterfly species richness and composition. I monitored butterfly communities during the first two years after implementation in five to seven burn units and controls in the watershed. To estimate baseline spatial and temporal variation in butterfly communities in the greater ecosystem, I also monitored butterflies in five untreated canyons outside the project area. Butterfly species richness and butterfly species composition (measured as community similarity) did not differ significantly between burn units and controls. Geographic location had statistically significant effects on species richness. Butterfly species composition of individual locations varied over time, as did the magnitude of that variation. These results emphasize that standardized, repeatable monitoring protocols are vital for evaluating the effects of experimental management treatments and for predicting and assessing the effects of future management strategies and environmental changes. PMID- 11029119 TI - Impact of Foot Traffic from Military Training on Soil and Vegetation Properties. AB - The impact of military training activities (primarily foot traffic) on soils and vegetation was assessed at the United States Air Force Academy, Colorado, USA. In May-June 1998 after 2 years of intensive training use, mean bulk densities of the top 6 cm of soil in the high-use site (1.37 g/cm(3)) and moderate-use site (1.30 g/cm(3)) were significantly different from bulk density of the reference site (1.04 g/cm(3)). Mean infiltration rates on the high use site (0.63 cm/min) and moderate use site (0.67 cm/min) were significantly different from the infiltration rate on the reference site (3.83 cm/min). Soil water holding capacities of the three sites were not significantly different. Descriptive comparisons of total aboveground biomass and litter indicated a 68% decrease in total aboveground biomass and a 91% decrease in litter when the high-use site was compared to the reference site. Using the Universal Soil Loss Equation, an estimated soil erosion rate for the reference plot (0.07 tons/ha/yr) was 30 times less than the erosion rate for the high use plot in the center of the basic cadet training encampment area (2 tons/ha/yr) and between 7 and 6 times less than the moderate use plot and the high use plot on the edge of the encampment area (0.5 and 0.4 tons/ha/yr, respectively). Therefore, training use appears to adversely affect bulk density, infiltration, total aboveground biomass, litter, and erosion. Without implementation of restoration practices, further site degradation is likely. PMID- 11029120 TI - Who should provide anaesthesia? PMID- 11029121 TI - Non-physician anaesthetists: can we agree on their role in Europe? PMID- 11029122 TI - Epidural analgesia-anaesthesia in obstetrics. AB - Epidural analgesia is the most effective and innocuous technique for obstetrics. Pain relief is its main indication but maternal diseases that might be decompensated by labour and delivery are also accepted indications. Low doses of long-acting local anaesthetics alone or in combination with low doses of fentanyl or sufentanil provide good quality analgesia and are safe for mother and fetus. Test doses in parturients lack sufficient specificity and sensitivity for detecting inadvertent intravascular injection, and subarachnoid migration of the catheter is possible at any time during the procedure. Therefore, every injection must be considered as a test dose and only fractionated injections must be made. Epidural block to T10 is needed for labour and to level T4 for Caesarean section. Maintenance of the block with a continuous infusion, or patient-controlled epidural analgesia with a background continuous infusion, provides more stable analgesia than by intermittent injection. Technical difficulties, dural tap, bloody tap, hypotension and insufficient block are most frequent complications of epidural block in obstetrics. Excessive motor block prolongs the second stage of labour and increases the frequency for instrumental delivery and is therefore considered a complication. PMID- 11029123 TI - The volumetric fluid balance as a measure of fluid absorption during transurethral resection of the prostate. AB - To evaluate the usefulness of the volumetric fluid balance for indicating and quantifying fluid absorption during transurethral resection of the prostate, 62 patients showing fluid absorption on ethanol monitoring (control method) were selected from a series of 410 operations. The volumetric fluid balance, which was measured as the difference between the input and output of irrigating fluid with and without a correction for the blood loss, proved to be an unreliable clinical tool for measuring the absorption. It indicated that fluid absorption occurred only in 40 or 18 of the 62 patients, depending on whether a correction for blood loss was made or not, the volume being only 59% and 71% (median), respectively, of that obtained by the control method. The absorption averaged 1 L in the patients in whom the volumetric measurements did not indicate absorption. The incidence of symptoms of the 'transurethral resection syndrome' increased with the absorbed fluid volume only when measured by the control method. PMID- 11029124 TI - Acute pain management: analysis, implications and consequences after prospective experience with 6349 surgical patients. AB - An acute pain service (APS) was set up to improve pain management after operation. We attempted to reduce the length of stay in the intensive care unit (ICU) of patients undergoing major surgery and to improve their homeostasis and rehabilitation using a multimodal approach (pain relief, stress reduction, early extubation). Patient-controlled epidural analgesia (PCEA) was a keystone of this approach. If PCEA was not applicable, patients received patient-controlled intravenous analgesia (PCIA) instead. Brachial plexus blockade (BPB) was used for surgery of the upper limbs. A computer based documentation system was used to help evaluate prospectively (a) the quality of analgesia, (b) adverse effects and risks of the special pain management techniques, and (c) cost-effectiveness. Patients receiving PCEA (n = 5.602) received a patient-titrated continuous infusion into the epidural space of either bupivacaine 0.175% or ropivacaine 0.2%, with 1 microg sufentanil mL(-1) added, followed by patient-controlled boluses of 2 mL (lockout time 20 min). For patients receiving PCIA (n = 634) an initial bolus of 7.5-15 mg piritramide was given, and the subsequent bolus was 2 mg (lockout time 10 min). A continuous infusion of bupivacaine 0.25% was administered to patients receiving BPB (n = 113). The dose was titrated to a dynamic visual analogue scale (VAS) scores < 40. The mean treatment periods were: BPB = 4.33 days, PCEA = 5.6 days, PCIA = 5.0 days. In the case of PCEA, the quality of pain relief, vigilance and satisfaction were superior compared with the PCIA method, which resulted in greater sedation and nausea. Although personal supervision was higher for the PCEA-treated patients, cost analysis revealed final savings of Euro 91,620 for the year 1998 obviating the need for an ICU stay totalling 433 days. Provided that PCEA is part of a fast-track protocol employing early tracheal extubation and optimal perioperative management, the associated initial higher costs will be recouped by the benefits to patients of better pain relief after surgery and fewer days subsequently spent in the ITU. PMID- 11029126 TI - Comparison of cisatracurium-induced neuromuscular block at the masseter and adductor pollicis muscle. AB - Adequate relaxation of the masseter muscle is important during endotracheal intubation and for the patency of a patient's airway during recovery from anaesthesia. We evaluated onset and recovery from cisatracurium-induced neuromuscular block at the masseter and adductor pollicis muscles. Thirty patients were randomly allocated to receive either 0.1 or 0.15 mg kg(-1) cisatracurium. The evoked response was measured at both muscles using acceleromyography. Onset time was significantly shorter at the masseter muscle than at the adductor pollicis (0.1 mg kg(-1) cisatracurium: 155+/-52 vs. 229+/-44 s; 0.15 mg kg(-1) cisatracurium: 105+/-24 vs. 174+/-35 s). Following 0.1 mg kg( 1) cisatracurium, recovery to a TOF-ratio of 0.7 was faster at the masseter compared to the adductor pollicis (P < 0.05). In the 0.15 mg kg(-1) cisatracurium group recovery of T1 to 75% of control and to a TOF-ratio of 0.7 occurred sooner at the masseter (P < 0.05). We conclude that onset and recovery from cisatracurium neuromuscular block occurs more rapidly at the masseter than at the adductor pollicis. It appears unlikely that residual paralysis is present at the masseter once neuromuscular function at the adductor pollicis has completely recovered. PMID- 11029125 TI - Double-blind randomized study of tramadol vs. paracetamol in analgesia after day case tonsillectomy in children. AB - Fifty children (2-9 years) scheduled for tonsillectomy were enrolled in a double blind randomized prospective study to compare postoperative analgesia provided with propacetamol/paracetamol (acetaminophen) or tramadol. A standard anaesthetic technique was used; each patient received sufentanil 0.25 microg kg(-1) intravenously followed with either i.v. propacetamol 30 mg kg(-1) or tramadol 3 mg kg(-1) before surgical incision. For postoperative analgesia, each child received either tramadol drops (2.5 mg kg(-1)) or paracetamol (acetaminophen) suppositories (15 mg kg(-1)), 6 and 12 h after surgery the first day and three times a day during postoperative days 2 and 3. This dosage of paracetamol is lower than the current recommended dosage, which is 40 mg kg(-1) loading dose followed by 20 mg kg(-1) 8 h(-1). Rescue medication consisted of i.v. diclofenac (1 mg kg(-1)) during the first six postoperative hours and oral ibuprofen (6-9 mg kg(-1)) afterwards. Postoperative pain scores (Children's Hospital of Eastern Ontario Pain Scale) in recovery, numerical pain scale in the ward and at home, and rescue analgesic use were significantly lower in the tramadol group. No serious adverse effects were observed. PMID- 11029127 TI - Ballotability of cuff to confirm the correct intratracheal position of the endotracheal tube in the intensive care unit. AB - The cuff ballotability method was used in 120 adult patients to confirm the correct depth of insertion of the endotracheal tube after tracheal intubation. The correct tube position was assumed when the cuff of the endotracheal tube could be felt to distend over the suprasternal notch when the pilot balloon was squeezed and the pilot balloon was felt to distend when pressure was applied over the suprasternal notch. Chest radiography was performed later to confirm the position of the endotracheal tube. In all patients the tip of the endotracheal tube was found to be in the desired position, i.e. 3-7 cm from the carina--the level of T3-T4 vertebrae. We concluded this technique to be a simple and reproducible way to confirm the correct depth of insertion of endotracheal tubes. PMID- 11029128 TI - Oxygen delivery failure resulting from interference with a Bain breathing system. AB - A critical incident is described where the expiratory (scavenging) port of an obsolete version of a Bain-type breathing system valve assembly was inadvertently connected to the common gas outlet of an anaesthetic machine. This resulted in the patient being deprived of a supply of fresh gas. This misconnection was made possible by adding a plastic connector to the valve scavenging port and subsequent wrongful misconnection. The patient fortunately suffered no harm. The case highlights the danger of equipment that has been subjected to unauthorized interference. PMID- 11029129 TI - High subarachnoid block for severe bronchospasm. PMID- 11029130 TI - Performance of US hospitals on recommended screening and immunization practices for pregnant and postpartum women. AB - OBJECTIVE: Recommendations by most national advisory committees on immunization include evaluating all pregnant women for chronic hepatitis B virus infection and immunity to rubella. It is recommended that all pregnant women be screened for hepatitis B surface antigen during an early prenatal visit and that rubella vaccine be administered in the postpartum period to women not known to be immune. This study determined the extent to which hospitals with labor and delivery services adhere to these recommendations. METHODS: We conducted a mail survey of a stratified random sample of all US medical-surgical hospitals to (1) determine the proportion of hospitals with hepatitis B screening policies and rubella immunization programs and (2) identify significant factors associated with the presence of these policies and programs. Hospitals were stratified by number of beds (<100, 100-499, and > or =500) and affiliation with a medical school. RESULTS: Of 986 institutions surveyed, 858 (87%) responded. Of these, 635 (74%) were labor and delivery hospitals. Approximately half of these (51%) had hospital policies related to screening pregnant women for the hepatitis B surface antigen. Twenty-one percent had rubella immunization programs for postpartum women. Only 14% of labor and delivery hospitals were in full compliance with published recommendations for hepatitis B surface antigen screening and rubella postpartum vaccination. Hospitals were more likely to be compliant if they had more than 100 beds, were private rather than public institutions, were affiliated with a medical school, and were in states with laws regarding hepatitis B surface antigen screening of pregnant women. CONCLUSIONS: Almost half, and more than three quarters, of hospitals were not in compliance with hepatitis B screening and rubella postpartum immunization recommendations, respectively. Hospitals should develop and implement policies for these preventive services. PMID- 11029131 TI - Ventilator-associated pneumonia in very low-birth-weight infants at the time of nosocomial bloodstream infection and during airway colonization with Pseudomonas aeruginosa. AB - PURPOSE: To study retrospectively the incidence of ventilator-associated pneumonia (VAP) at the time of Pseudomonas aeruginosa nosocomial bloodstream infection (BSI) and at the time of P aeruginosa airway colonization. MATERIALS AND METHODS: Fifteen very low-birth-weight infants who had P aeruginosa BSI and 33 others who did not but who had P aeruginosa airway-colonization were studied. We correlated clinical data, blood cultures (BCs), and tracheal cultures (TCs) with radiologic findings from radio-graphs taken within 2 days before, the day of, and 1 day after BCs or TCs were first positive for P aeruginosa. Chest radiographs were graded by using semiquantitative scores for bronchopulmonary dysplasia and for pneumonia. RESULTS: Mean birth weight, gestational age, and age when BC or TC became positive were similar for patients with BSI and colonization. At the time of BSI, 2 infants had airway colonization with P aeruginosa; the TCs of the remaining 13 grew P aeruginosa as a new pathogen. Thirteen of 15 patients with BSI, but none of 33 infants with colonization, died within 2 days of positive BC. VAP was diagnosed in 13 of 15 patients with BSI and in 3 of 33 infants with colonization. CONCLUSION: Mechanically ventilated very low-birth-weight infants whose TCs yield P aeruginosa but whose BCs remain negative infrequently have VAP are presumed airway-colonized and are expected to survive. Conversely, VAP is likely to be found when BCs and TCs simultaneously grow P aeruginosa, and high mortality is anticipated. PMID- 11029132 TI - Effect of hand sanitizer use on elementary school absenteeism. AB - BACKGROUND: Several studies have indicated a connection between handwashing and illness-related absenteeism in school settings. The difficulty of ensuring consistent and effective handwashing among student populations has also been noted. The purpose of this study was to assess the effectiveness of the use of an alcohol gel hand sanitizer in the classroom to help decrease the illness-related absentee rate for elementary school students. METHODS: This study involved 5 individual school districts, 16 individual schools, and more than 6000 students in Delaware, Ohio, Tennessee, and California. Individual schools in each district were paired into product and control groups. In the product group schools, an alcohol gel hand sanitizer was used by the students and staff when entering and leaving the classroom. Absenteeism due to infection was recorded, and the data were statistically analyzed. RESULTS: The overall reduction in absenteeism due to infection in the schools included in this study was 19.8% for schools that used an alcohol gel hand sanitizer compared with the control schools (P <.05). Data from the school system with the largest teacher population (n = 246) showed that teacher absenteeism decreased 10.1% (trend) in the schools where sanitizer was used. CONCLUSION: Elementary school absenteeism due to infection is significantly reduced when an alcohol gel hand sanitizer is used in the classroom as part of a hand hygiene program. PMID- 11029133 TI - Microbiologic contamination study of nebulizers after aerosol therapy in patients with cystic fibrosis. AB - BACKGROUND: To evaluate the contamination of delivery systems after an aerosol therapy session in patients with cystic fibrosis who have chronic Pseudomonas aeruginosa infection. METHODS: Fifty-three patients with cystic fibrosis were enrolled in the study from March 1996 to June 1997. All patients were age 7 years or older and had P aeruginosa infection. They also had been treated with recombinant deoxyribonuclease and were capable of producing sputum for culture. RESULTS: Nine devices were excluded for the study. A total of 44 nebulizers were included: 37 from patients with P aeruginosa colonization with a count of 10(6) colony-forming units/mL or more and 7 with a count of between 10(5) colony forming units/mL and 10(6) colony-forming units/mL. CONCLUSION: This study demonstrates that in the absence of cleaning, nebulizers of patients with cystic fibrosis who are infected with P aeruginosa are likely to be contaminated by a pathogenic flora. PMID- 11029134 TI - Latex gloves use and symptoms in health care workers 1 year after implementation of a policy restricting the use of powdered gloves. AB - BACKGROUND: Latex sensitization related to glove use is a complex problem, and glove use policies that restrict exposure to powdered latex gloves alone may not provide adequate safety. This study explored health care workers' (HCWs') latex glove use and reports of related health symptoms 1 year after implementation of the latex glove replacement policy. METHODS: Two cross-sectional surveys. One was a population-based survey of HCWs at the institution (National Surveillance System for Hospital Health Care Workers); the second, a detailed survey of HCWs stratified by exposure to latex gloves (Latex Symptom Survey). RESULTS: The prevalence of symptoms of dermatitis reported by latex glove users was 40.3% (National Surveillance System for Hospital Health Care Workers) and 50.0% (Latex Symptom Survey). Symptom reports increased as exposure to latex gloves increased (pairs used per day, occupations with more glove use, working in higher exposure areas, more tenure, and more hours of use per day). Aerosol symptoms and urticaria related to latex glove use were reported much less frequently than were rash, chapping, itching, or redness. HCWs, even those with skin symptoms, continued to choose latex gloves in more than 80% of the cases. CONCLUSIONS: Stepwise preplacement evaluation at employee health clinics is recommended to identify and protect employees sensitized to latex. The medical community needs more understanding and education about latex gloves, latex sensitization, and available alternatives. PMID- 11029135 TI - Determinants of nurses' adherence to universal precautions for venipunctures. AB - OBJECTIVES: The aim of this study was to predict and explain nurses' adherence to Universal Precautions (UPs) when performing venipunctures. METHODS: Data were gathered from 156 registered nurses working at a regional hospital. A self administered questionnaire assessing the psychosocial variables (intention, attitude, subjective norm, perceived control, etc) was completed at baseline, and behavior was self-reported 3 months later. RESULTS: The regression of intention on the variables yielded an adjusted R(2) of 0.68, with perceived barriers (beta =.62, P <.001), social norm (beta =.17, P <.01), and personal normative belief (beta =.19, P <.01) being the significant variables. With respect to the 3-month follow-up, 28% of the variance associated with UPs adherence was explained by intention (beta =.37, P <.001) and perceived behavioral control (beta =.23, P <.05). Moreover, high (n = 116) and low intenders (n = 40) differed on several normative beliefs (P =.0003) and perceived barriers (P =.0001). CONCLUSIONS: It is suggested that perception of control, assessed either globally or by means of a belief-based measure, is a key factor in adherence. Specifically, the perceived difficulties associated with adherence to UPs appear to be related to a nurse's training on UPs and to the existence of suboptimal working conditions. PMID- 11029136 TI - Using peer feedback to improve handwashing and glove usage among Thai health care workers. AB - BACKGROUND: Without protective practices such as Universal Precautions, health care workers are at substantial risk for bloodborne infection, especially in areas such as Thailand with high prevalence of HIV infection. The purpose of this study was to evaluate the effectiveness of a peer feedback program (PFP) on handwashing and glove wearing (HW/GW) among Thai health care workers. METHODS: Subjects (N = 91) were randomly assigned to receive PFP versus no treatment. By using a checklist, peer observers rated HW/GW compliance in their coworkers during patient care. For 1 month, the investigator posted a report of compliance behaviors from each 3 days of observations. HW/GW was also assessed by the investigator by direct observation at 1 month before the intervention, during the intervention period, and 1 month after the intervention. RESULTS: Baseline HW/GW rates for the PFP and control groups were 49.2% and 61.5%, respectively. The PFP group had a significantly higher adjusted compliance rate than the control group during the intervention period (P =.0001). However, there was no significant difference in the compliance scores obtained 1 month after the intervention. CONCLUSIONS: The PFP was effective during the intervention period, but there was no retention of effect. Therefore, adjunct methods should be sought to promote retention of effect. PMID- 11029137 TI - Evaluation of a novel, rapid-acting, sterilizing solution at room temperature. AB - Test data are presented for a novel chemical germicide formulation capable of sterilizing reusable medical devices in 30 minutes at 20 degrees C in an open tray. The tests conducted with this rapid-acting sterilizing solution (RSS) included sporicidal, mycobactericidal, and virucidal studies performed in accordance with Association of Official Analytical Chemist International or Environmental Protection Agency published guidelines, by using RSS stressed for as long as 7 days. Sporicidal assays were performed at 20 degrees C with a 30 minute exposure time by using both Clostridium sporogenes and Bacillus subtilis spores dried on porcelain penicylinders or suture loops (n = 60 carriers per treatment). For comparison, identical carriers were exposed to a commercial glutaraldehyde-based sterilizing solution stressed to a maximum-use time of 14 days and exposed per manufacturer's requirements (10 hours at 25 degrees C). The RSS sterilized 100% of the carriers of both spore types. The glutaraldehyde solution demonstrated 100% sterilization of C sporogenes -treated carriers but had difficulty sterilizing B subtilis spore-laden carriers (ie, no sterilization of suture loops and only 57% sterilization of porcelain penicylinders). Similarly, Mycobacteria bovis and selected fungal and viral agents were exposed to stressed solution for 5 minutes or less at 20 degrees C. In each case, the resulting log decrease in viable microorganisms significantly supported a claim for rapid high-level disinfection. Based on these data, RSS demonstrates high level disinfection in 5 minutes and sterilization in 30 minutes at 20 degrees C. PMID- 11029138 TI - Acinetobacter baumanii-associated arterial line infection. PMID- 11029139 TI - Rotavirus outbreak on a pediatric oncology floor: possible association with toys. AB - BACKGROUND: Several outbreaks of rotavirus gastroenteritis have occurred in hospitals and day care centers. In the spring of 1997, an outbreak of rotavirus occurred on our pediatric unit. Aggressive infection control measures were instituted, and potential lapses in infection control were assessed. METHODS: Memorial Sloan-Kettering Cancer Center is a 434-bed cancer hospital in New York City. The pediatric unit is a 42-bed ward with both bone marrow transplant patients and non-bone marrow transplant oncology patients. Nosocomially acquired rotavirus was defined as diarrhea, vomiting, or gastrointestinal upset with onset 48 hours or more after hospital admission, accompanied by a positive enzyme immunoassay for rotavirus antigen. RESULTS: Between February 24 and April 4, 1997, 8 patients on the pediatric unit had nosocomial rotavirus. Aggressive infection control measures were instituted. Patients with rotavirus were cohorted and placed on contact precautions (strict handwashing, gloves, and gown). Investigation by the infection control team revealed that communal toys in the playroom were not being cleaned according to the weekly protocol. CONCLUSIONS: An outbreak of nosocomial rotavirus occurred on our pediatric oncology unit. Shared toys may have served as fomites in the transmission of rotavirus. PMID- 11029140 TI - Infection control in cystic fibrosis: practical recommendations for the hospital, clinic, and social settings. PMID- 11029141 TI - Macular rash following influenza vaccine. PMID- 11029142 TI - Environmental protection in hospital infection control. PMID- 11029144 TI - Pathogenic bacteria on personal pagers. PMID- 11029145 TI - Cardiovascular malformations: epidemiologic study presages endothelial cell origin. PMID- 11029146 TI - Anti-HIV therapy during pregnancy: risk-benefit ratio. PMID- 11029147 TI - Developmental toxicity of the HIV-protease inhibitor indinavir in rats. AB - BACKGROUND: Indinavir is an antiviral agent used for the treatment of HIV infection. We studied its developmental toxicity in rats. METHODS: Pregnant animals were treated orally with 500 mg indinavir/kg body weight (bw) from day 6 to 15 of gestation (once daily) or from day 9 to 11 (twice daily). Fetuses were evaluated for external and skeletal anomalies on day 21 of gestation. In addition, 19 rats were treated from day 9 of gestation to day 24 postnatally with 500 mg indinavir/kg bw once daily; a control group of 17 rats was treated with the vehicle accordingly. Developmental landmarks were recorded. Sixteen offspring each were studied on postnatal days 7, 14, 21, and 35 for hepatic enzyme activity. Liver tissue was examined by electron microscopy. RESULTS: Fetal examination on day 21 of pregnancy showed no treatment-related effects on number, weight, and viability of the fetuses; however, an increased incidence was noted in the supernumerary ribs and variations of the vertebral ossification centers in both indinavir-treated groups. Postnatal evaluation showed delayed fur development, eye opening, and descensus testis. The most striking finding was unilateral anophthalmia, observed in 7 pups (3%) from 2 out of 19 litters exposed to indinavir, but not in controls. Only minor changes in hepatic monooxygenase activities occurred in dams. Electron microscopy of liver samples showed hepatocellular inclusions of lipids and myelin figure-like structures in maternal livers and infiltration with granulocytes in offspring livers. CONCLUSIONS: Further studies on reproductive toxicity, including combinations of three or more antiretroviral agents as used therapeutically, are needed to determine the hazards of such a treatment. PMID- 11029148 TI - A history of the Teratology Society. AB - BACKGROUND: The 39-year history of the Teratology Society is reviewed. An abbreviated history is outlined in table form, along with listings of the Warkany Lectures, the postgraduate courses, and officers of the Society. METHODS: A year by-year description of the events, including the scientific and social content of the annual meetings and changes in the business of the Society, is given, in many cases using comments from the past presidents. The valuable and unique diversity of the members is discussed and illustrated, presenting the disciplines and main research area of the presidents. The number of submitted abstracts and the various categories are tabulated, averaging the number and type over four periods. Within the past 10 years, a significant increase in the number of abstracts dealing with epidemiology and developmental biology is evident. The Society's development is compared with that of a human, and the question is asked: Have we reached the maturational stage of old age or senescence, or is the Society still maturing gracefully? This question needs further discussion by all the members. RESULTS: During the past 40 years, we have developed the scientific basis to prevent birth defects caused by rubella, alcoholism, and folate deficiency, as well as many other prenatal exposures. CONCLUSIONS: We must now engage in the political battles to obtain the resources needed to conduct further research and to implement the prevention programs, as well as to provide care and rehabilitation for persons with birth defects. PMID- 11029149 TI - alpha(1)-Adrenergic stimulation perturbs the left-right asymmetric expression pattern of nodal during rat embryogenesis. AB - BACKGROUND: Normal development of the left/right (L/R) body axis leads to the characteristic sidedness of asymmetric body structures, e.g., the left-sided heart. Several genes are now known to be expressed with L/R asymmetry during embryogenesis, including nodal, a member of the transforming growth factor-beta (TGF-beta) family. Mutations or experimental treatments that affect L/R development, such as those that cause situs inversus (reversal of the sidedness of asymmetric body structures), have been shown to alter or abolish nodal's asymmetric expression. METHODS: In the present study, we examined the effects on nodal expression of alpha(1)-adrenergic stimulation, known to cause a 50% incidence of situs inversus in rat embryos grown in culture, using reverse transcription-polymerase chain reaction assay and whole-mount in situ hybridization assay. RESULTS: In embryos cultured with phenylephrine, an alpha(1) adrenergic agonist, nodal's normal asymmetric expression only in the left lateral plate mesoderm was altered. In some treated embryos, nodal expression was detected in either the left or right lateral plate mesoderm. However, most treated embryos lacked lateral plate mesoderm expression. In addition, the embryos that did show expression were at a later stage than when nodal expression is normally found. CONCLUSIONS: Our results demonstrate that alpha(1)-adrenergic stimulation delays the onset and perturbs the normal asymmetric pattern of nodal expression. Either of these effects might contribute to situs inversus. PMID- 11029150 TI - Caffeine decreases the occurrence of cadmium-induced forelimb ectrodactyly in C57BL/6J mice. AB - BACKGROUND: Cadmium is a well-known animal teratogen. Caffeine is an alkaloid widely consumed by humans. Interactions between teratogens and nonteratogenic doses of other agents are becoming widely studied, as they may shed light on understanding mechanisms of teratogenicity or possible prevention of teratogenic effects. METHODS: C57BL/6JBK mice were injected intraperitoneally (ip) with cadmium sulfate (Cd) at 0, 1.00 (LDCd), 2.50 (MDCd), or 5.00 (HDCd) mg/kg, immediately followed by subcutaneous (sc) administration of 0 or 50 mg/kg caffeine (CAFF) on gestation day (GD) 9. Fetuses were examined on GD 18 for ectrodactyly and other gross morphological malformations. RESULTS: Amelioration of cadmium-induced forelimb ectrodactyly by CAFF was seen in both the high-dose cadmium (HDCd = 65.4%, HDCd+CAFF = 39.2%) and medium-dose cadmium (MDCd = 46.2%, MDCd+ CAFF = 20.8%) treatment groups (P < 0.025). Bilateral expression of ectrodactyly was also decreased in the presence of caffeine. A statistically significant reduction in Cd-induced abnormalities, including: eye, abdominal, and other skeletal defects, was not seen with caffeine addition, although they did trend downward in the caffeine-supplemented groups. Litter size, fetal weight, fetal mortality, and dam weight also were not affected by co-treatment with caffeine. CONCLUSIONS: This study provides evidence that a subteratogenic dose of caffeine can ameliorate cadmium-induced forelimb ectrodactyly in the Cd-sensitive C57BL/6J inbred mouse strain. PMID- 11029151 TI - Teratogen update: maternal myasthenia gravis as a cause of congenital arthrogryposis. AB - BACKGROUND: Arthrogryposis multiplex congenita (AMC) is defined as nonprogressive congenital contractures that generally result from lack of fetal movement in utero. AMC is a feature of many congenital disorders caused by genetic, environmental, or other factors. One rare cause of AMC is maternal myasthenia gravis (MG). This is an autoimmune disorder, caused by antibodies to the nicotinic acetylcholine receptor (AChR), and resulting in weakness of voluntary muscles. In 10-15% of babies born to MG mothers, transient signs of MG are noted after placental transfer of anti-AChR antibodies. In a few cases, AMC predominates. METHODS: We review the role of antibodies to AChR in MG and in AMC associated with maternal antibodies to AChR. RESULTS: In anti-AChR antibody associated AMC, fetal or neonatal death is common; other deformities or CNS abnormalities are common as well. The condition usually recurs in each pregnancy unless the mother is treated for MG, but some mothers are asymptomatic. The maternal antibodies cross the placenta and block the function of the fetal isoform of the AChR leading to fetal paralysis. Injection of maternal plasma into pregnant mice results in AMC in mouse fetuses. Some women with recurrent AMC in their babies have no detectable anti-AChR suggesting the presence of antibodies to other fetal muscle or neuronal proteins. CONCLUSIONS: Although rare, anti-AChR associated AMC is potentially treatable and can be diagnosed by a routine antibody test. The mouse model can be used to investigate the role of these and other maternal antibodies in causing congenital conditions. PMID- 11029152 TI - Are oral clefts a consequence of maternal hormone imbalance? evidence from the sex ratios of sibs of probands. AB - BACKGROUND: The causes of oral clefts (cleft lip with or without cleft palate, CL/P, and cleft palate alone, CP) have not been established. However, maternal intrauterine hormone profiles have been suspected of being involved. There is now substantial evidence that maternal hormone concentrations around the time of conception partially control the sexes of offspring. It is possible that the hormone profiles that control sex of offspring share features of the profiles suspected of causing clefts. This can be tested by examining the sex ratios (proportions male) of the unaffected sibs of probands. If these sex ratios are skewed in the same direction as that of probands, that suggests, ex hypothesi, maternal hormonal involvement in the causation of clefts. METHODS: Accordingly, a search was made for data on the sex ratios of the unaffected sibs of probands with clefts. For reasons given in the text, this search was informal rather than based on electronic data retrieval systems. Nine papers were located giving sex ratios of sibs of probands with CL/P and CP. RESULTS: Published data suggest that the sibs of probands with CL/P have a significantly higher sex ratio than the sibs of probands with CP. Thus the sib sex ratios are skewed in the same direction as those of the probands themselves. In other words, parents (mothers) of CL/P patients apparently have a tendency to produce boys, and parents of CP patients to produce girls. CONCLUSION: Accordingly, it is suggested that maternal hormone profiles may partially explain the unusual sex ratios (of probands and their sibs), as well as the malformations. PMID- 11029153 TI - Hydroxylamine moiety of developmental toxicants is associated with early cell death: a structure-activity analysis. AB - BACKGROUND: Cellular debris, an indicator of cell death, appears in limb buds of gestational day 12 rabbit embryos 4 hr after either a subcutaneous injection of hydroxyurea to pregnant rabbits or an injection of hydroxyurea into the exocoelomic cavities of the embryos. This episode of early cell death appears to be central to the teratogenic action of hydroxyurea. Several chemicals that are structurally related to hydroxyurea, and that possess a terminal hydroxylamine moiety (-NHOH), also produce limb abnormalities. METHODS: To investigate whether the hydroxylamine moiety is responsible for early cell death and, therefore, is likely to be associated with teratogenesis, five structurally related hydroxylamine-bearing chemicals (hydroxylamine hydrochloride, N methylhydroxylamine hydrochloride, hydroxyurea, acetohydroxamic acid, and hydroxyurethane) were administered at equimolar doses to rabbits either by subcutaneous (8.55 mmol/kg) or intracoelomic (2.66 micromol/embryo) injection on gestational day 12. Five additional chemicals, structurally similar to the hydroxylamine-bearing compounds, but possessing a terminal amino group (-NH(2)) (ammonium hydroxide, methylamine, urea, acetamide, and urethane), were tested at equimolar or higher doses by an identical protocol. In a subsequent experiment, the antioxidant propyl gallate (3.0 mmol/kg or 1.30 micromol/embryo) was co administered with the hydroxylamine-bearing compounds to determine its effect on early cell death. Embryos were harvested 4 or 8 hr after treatment and analyzed by light microscopy. RESULTS: Cellular debris was obvious in forelimb buds from embryos treated with the hydroxylamine-bearing compounds; however, none of the amino compounds produced an early episode of embryonic cell death. In all cases, the antioxidant propyl gallate prevented or delayed the early episode of cell death observed after treatment with the hydroxylamine-bearing compounds. CONCLUSIONS: These results are consistent with the concept that the rapidly occurring embryonic cytotoxicity induced by hydroxylamine-bearing compounds involves a free radical mechanism that requires the presence of a terminal hydroxylamine group for initiation. PMID- 11029154 TI - Acardiac anomaly spectrum. AB - BACKGROUND: Acardiac anomaly spectrum is a rare congenital malformation found in monozygotic twin pregnancy. Besides the absence of heart, the condition is associated with variable grades of developmental disruption. Thus, no two cases are similar. METHODS: This case report is based on physical examination and autopsy findings. RESULTS: The twin had acardia and partial development of head and face. There was complete absence of upper extremities. CONCLUSIONS: The twin reversed arterial perfusion (TRAP) theory is the most accepted etiology of the disorder. Normally, the cephalic pole is the most severely affected, being most distal to the retrograde perfusion. In acardia, partial development of head, face, and brain is usually associated with the development of the upper extremities. However, in the present case, there was extensive cephalic development in the absence of upper extremity development. PMID- 11029155 TI - Foreword PMID- 11029156 TI - Optimization of multiple-isocenter treatment planning for linac-based stereotactic radiosurgery. AB - OBJECTIVE: Computer-assisted treatment planning for linac-based radiosurgery is still an open research problem, especially for multiple-isocenter procedures, primarily due to its high complexity and computational requirements. This paper focuses on the optimization of multiple-isocenter treatment planning for linac systems, and addresses several important issues associated with multiple isocenters, such as dose conformality, homogeneity, and optimization of isocenter position and dose. METHODS: The key idea behind our approach is that the desired dose distribution can be decomposed into a number of fundamental components. In the current paper, an analytical form, the so-called Ellipsoidal Dose Distribution Estimation (EDDE) model, represents each component. We establish ways (arc configurations) to achieve such ellipsoidal doses of arbitrary position, orientation, and size. Since the EDDE model is described by relatively few parameters, it allows very quick estimation of the dose distribution corresponding to a particular isocenter and thus makes the optimization of isocenter position very efficient. It is further used in a framework for optimal treatment planning, in which a number of ellipsoidal dose distributions, each corresponding to a different isocenter, are optimally placed to cover the target while sparing healthy tissue. RESULTS: The general ellipsoidal dose distribution of linac-based radiosurgery is summarized as a mathematical model with the aid of supporting experiments. Comparisons between the EDDE-optimized and clinically implemented plans are made, revealing the superior performance of the former. In addition, a dramatic reduction in planning time is achieved using the EDDE model. CONCLUSION: The proposed EDDE model is a useful and effective dose model in multiple-isocenter treatment planning for linac-based radiosurgery. PMID- 11029157 TI - 3D dynamic body surface sensing and CT-body matching: a tool for patient set-up and monitoring in radiotherapy. AB - Recent advances in radical broad-beam conformal radiotherapy for disease at non rigid sites suggest that the treatment room set-up of patients should be performed using the CT-plan digital body surface as a reference standard. In effect, the CT-plan surface is a "virtual shell" and should be used as such. To do this requires 3D body surface information to be captured live in the treatment room and then manipulated for comparison with the CT virtual shell. To address this situation, a near real-time prototype opto-electronic dynamic 3D surface sensor has been developed and equipped with a novel software utility for visualizing and matching the disparate surfaces arising from planning and treatment environments. Using the CT virtual shell and height-maps produced by the sensor, a two-phase 3D positioning strategy for patient set-up and monitoring is described. Phase one is an image-guided manual "approach". Phase two is automated "docking" using simulated annealing to determine optimum set-up corrections. The concepts and implementation are illustrated using archival patient data and an anthropomorphic phantom "patient" in a treatment room environment. PMID- 11029158 TI - Patient set-up using portal images: 2D/2D image registration using mutual information. AB - OBJECTIVE: Conformal radiation therapy requires accurate patient set-up for each fraction delivery. Electronic portal imaging devices allow the acquisition of portal images just before and even during dose delivery. However, the quantitative interpretation of these images in determining and correcting the patient's position remains uncertain, and automated methods are therefore being developed. Such methods must be usable for the different radiation therapy techniques. They must be robust and as automated as possible for use in clinical routines. This work was undertaken to establish the feasibility of 2D/2D registration for portal/portal and portal/simulator images in radiotherapy. MATERIALS AND METHODS: This paper describes an automated method based on the combination of calibration algorithms and pixel-based registration algorithms. We present experiments with the different imaging techniques, some of which use a phantom with and without a gold standard. Preliminary results obtained using patient data are also presented and discussed. RESULTS: The results obtained with a phantom demonstrated that this automated method for 2D/2D registration is fast, accurate, and robust, even in the case of blurred images for small treatment fields. CONCLUSIONS: Mutual information is a feasible method for 2D/2D portal/portal and portal/simulator image registration in radiotherapy. PMID- 11029159 TI - Robotic motion compensation for respiratory movement during radiosurgery. AB - Tumors in the chest and abdomen move during respiration. The ability of conventional radiation therapy systems to compensate for respiratory motion by moving the radiation source is inherently limited. Since safety margins currently used in radiation therapy increase the radiation dose by a very large amount, an accurate tracking method for following the motion of the tumor is of the utmost clinical relevance. We investigate methods to compensate for respiratory motion using robotic radiosurgery. Thus, the therapeutic beam is moved by a robotic arm, and follows the moving target tumor. To determine the precise position of the moving target, we combine infrared tracking with synchronized X-ray imaging. Infrared emitters are used to record the motion of the patient's skin surface. A stereo X-ray imaging system provides information about the location of internal markers. During an initialization phase (prior to treatment), the correlation between the motions observed by the two sensors (X-ray imaging and infrared tracking) is computed. This model is also continuously updated during treatment to compensate for other, non-respiratory motion. Experiments and clinical trials suggest that robot-based methods can substantially reduce the safety margins currently needed in radiation therapy. PMID- 11029160 TI - Image-guided radiosurgery for the spine and pancreas. AB - A robotic image-guided radiosurgical system has been modified to treat extra cranial sites using implanted fiducials and skeletal landmarks to locate the treatment targets. The system has been used to treat an artero-venous malformation in the cervical spine, a recurrent schwannoma of the thoracic spine, a metastatic adenocarcinoma of the lumbar spine, and three pancreatic cancers. During each treatment, the image guidance system monitored the position of the target site and relayed the target coordinates to the beam-pointing system at discrete intervals. The pointing system then dynamically aligned the therapy beam with the lesion, automatically compensating for shifts in target position. Breathing-related motion of the pancreas lesions was managed by coordinating beam gating with breath-holding by the patient. The system maintained alignment with the spine lesions to within +/- 0.2 mm on average, and to within +/- 1 mm for the pancreatic tumors. This experience has demonstrated the feasibility of using image-guided robotic radiosurgery outside the cranium. PMID- 11029161 TI - Ultrasound-based stereotactic guidance in prostate cancer--quantification of organ motion and set-up errors in external beam radiation therapy. AB - OBJECTIVE: A mobile transabdominal ultrasound-based targeting system (BAT(R)) has been developed which can stereotactically localize the position of the prostate each treatment day and directly integrate this information into the treatment planning system. Daily target verification facilitates a marked reduction in planning treatment margins by correcting potential organ-motion and set-up errors. Previous studies have been performed to establish the precision of ultrasound localization. This report quantifies the magnitude of the patient isocenter shift parameters encountered during clinical implementation of this system. MATERIAL AND METHODS: After five weeks of conformal external beam radiation therapy, 54 patients underwent a second CT simulation. Prostate-only fields based on this scan were created with no PTV margin beyond the CTV. For each of the final conedown treatments (2-4 fractions), patients underwent ultrasound-based stereotactic prostate localization at the treatment machine. The portable system, which electronically imports the CT simulation target-contour and isocenter information, is situated adjacent to the treatment couch. Transverse and sagittal suprapubic ultrasound images are captured, and the system electronically couples this data to the baseline isocenter. The CT contours are maneuvered in three dimensions by a touch-screen menu to visually overlay the ultrasound images. The system then displays the three-dimensional (3D) couch shifts required to produce field alignment. RESULTS: One hundred and eighty-nine daily ultrasound prostate position shifts were recorded for 54 patients. The isocenter field misalignment between the baseline CT and ultrasound ranged from 26.8 to 33.8 mm in the anterior/posterior (A/P) dimension, -10.2 to 30.9 mm in the lateral dimension, and -24.6 to 9.0 mm in the superior/inferior (S/I) dimension. The corresponding directed average disagreements were -3.0 mm (SD 8.3 mm) A/P, 1.86 mm (SD 5.7 mm) lateral, and -2.6 mm (SD 6.5 mm) S/I. The magnitudes of undirected misalignments were frequently larger than 5 mm (51% of A/P, 31% of lateral, and 35% of superior measurements) and oftentimes larger than 10 mm (21% of A/P, 7% of lateral, and 12% of superior measurements). CONCLUSIONS: Organ motion and set-up uncertainties limit optimization of 3D treatment planning by expanding the width of PTV margins required to ensure target coverage. Transabdominal ultrasound-based stereotactic guidance is a safe and direct method for correcting patient positioning. Our experience with the BAT system in a large cohort of prostate cancer patients revealed that substantial daily isocenter corrections were encountered in a large percentage of cases. This data would suggest that daily clinical isocenter misalignments are greater than would be expected from published data on organ motion and set-up variations encountered in the study setting. PMID- 11029162 TI - Real-time opto-electronic verification of patient position in breast cancer radiotherapy. AB - OBJECTIVE: The clinical application of an opto-electronic system for real-time three-dimensional (3D) control of patient position in breast cancer radiotherapy is described. The specific features of the motion analysis technology (shape recognition of passive markers) are detailed, and the outcomes of its clinical use for quantitative position control and immobility verification of the thoracic irradiation field during breast cancer treatment are reported. MATERIALS AND METHODS: The position control system is based on the ELITEtrade mark opto electronic motion analyzer, which provides in real time the 3D coordinates of a set of passive markers (plastic hemispheres 3 mm in diameter) previously placed on selected landmarks on the patient's skin. The system-dedicated hardware performs marker recognition by means of 2D correlation of shape with a predefined marker modeling mask. This feature ensures a high accuracy, even with small marker dimensions, and successful analysis in a noisy environment (due to room light, reflexes, etc.). The patient repositioning control was based on a comparison between the current positions of the markers and a corresponding reference configuration. The resulting marker displacements were graphically displayed in real time for immediate control. This information was not provided to the operator as a repositioning tool. Instead, the kinematic data was stored for subsequent off-line analysis aimed at quantifying the different factors contributing to patient mis-positioning (initial repositioning errors, patient's breathing, and random movements) when conventional means for patient alignment (laser centering) and immobilization (casting techniques) are used. RESULTS: Clinical application of the system revealed median 3D localization errors for the directly controlled anatomical landmarks of around 4.5 mm. This value is proposed to represent the intrinsic accuracy of conventional laser-centering techniques in breast cancer radiotherapy, including the effects of patient body deformations. When the positional inaccuracies introduced by patients' respiration were also considered, the extent of the resulting 3D mis-positioning of the control points increased to median values of up to 8 mm. CONCLUSIONS: The reported clinical trial confirms the significant role that real-time opto-electronic motion analysis based on passive markers can have in augmenting the accuracy of patient repositioning and immobility verification in the radiotherapy of a non-rigid body area while also accounting for physiological movements. Evaluation of the data collected during each irradiation session for five patients provided valuable information concerning the optimization of the efficacy of traditional methods for patient centering and immobilization. PMID- 11029164 TI - High loading cellulose based poly(alkenyl) resins for resin capture applications in halogenation reactions PMID- 11029163 TI - Comprehensive survey of combinatorial library synthesis: 1999. PMID- 11029166 TI - Solid-phase synthesis of a heterocyclic ethylenediamine-derivatized library. PMID- 11029165 TI - Polymer-supported alpha-imino acetates. Versatile reagents for the synthesis of alpha-amino acid libraries. PMID- 11029167 TI - Solid phase synthesis of indinavir and its analogues. PMID- 11029168 TI - AutoChem: automated solution-phase parallel synthesis and purification via HPLC. PMID- 11029169 TI - Combinatorial screening of homogeneous catalysis and reaction optimization based on multiplexed capillary electrophoresis PMID- 11029170 TI - A novel approach to high-throughput quality control of parallel synthesis libraries. AB - Combinatorial chemistry is a powerful tool to enhance drug discovery efforts in the pharmaceutical industry. One type of combinatorial chemistry, parallel synthesis, is now widely used to prepare numerous compounds of structural diversity. A novel high-throughput method for quality control of parallel synthesis libraries has been developed. The method uses flow injection MS, for proof of structure and estimation of purity, and a novel direct injection CLND technique for quantitation of amount. Following the synthesis of a small molecule library, compounds analyzed using this technique were characterized by mass spectrometry, and an accurate concentration of the compound was assessed by CLND. Characterization of one compound is completed in 60 s, allowing for up to 1000 compounds to be analyzed in a single day. The data is summarized using pass/fail criteria using internally developed software. PMID- 11029171 TI - Rapid identification of substrates for novel proteases using a combinatorial peptide library. AB - Fluorogenic substrates for assaying novel proteolytic enzymes could be rapidly identified using an easy, solid-phase combinatorial assay technology. The methodology was validated with leader peptidase of Escherichia coli using a subset of an intramolecularly quenched fluorogenic peptide library. The technique was extended toward the discovery of substrates for a new aspartic protease of pharmaceutical relevance (human napsin A). We demonstrated for the first time known to us that potent fluorogenic substrates can be discovered using extracts of cells expressing recombinant enzyme to screen the peptide library. The straightforward and rapid optimization of protease substrates greatly facilitates the drug discovery process by speeding up the development of high throughput screening assays and thus helps more effective exploitation of the enormous body of information and chemical structures emerging from genomics and combinatorial chemistry technologies. PMID- 11029172 TI - Solution-phase synthesis of a 1,5-dialkylamino-2,4-dinitrobenzene library and the identification of novel antibacterial compounds from this library. AB - In this report we demonstrate that a 1,5-dialkylamino-2,4-dinitrobenzene small molecule library can be generated by a highly efficient solution-phase synthesis method. From this 2485-member library, a series of novel compounds with antibacterial activity were isolated. The significance of this report is that the synthetic scheme is extremely simple, with minimal number of liquid handling steps, and the solvents and reagents left in the final library preparation are fully compatible with cell-based assays. PMID- 11029174 TI - Combined application of analytical techniques for the characterization of polymer supported species AB - The combined application of a diverse range of analytical techniques is described for the complete analysis of polymer supported molecules. These techniques permit complete description of the FTIR, (1)H NMR, and (13)C NMR spectra. The comparison of supported bicyclo[2.2.2]octane derivatives with analogous species prepared using polymer supported reagents is made. PMID- 11029173 TI - 5-(hydroxymethyl)oxazoles: versatile scaffolds for combinatorial solid-phase synthesis of 5-substituted oxazoles. AB - A scheme combining the preparation of building blocks in solution followed by solid-phase combinatorial chemistry has been developed to side-chain diversify 5 (hydroxymethyl)oxazole scaffold (1) into aryl ethers, thioethers, sulfones, sulfonamides, and carboxamides. Protected heterocyclic scaffolds 2 were linked to the solid phase and N-terminal derivatized using active ester chemistry, providing chemset 4?1-4,1-4?. The free side-chain hydroxyl of 4 was smoothly converted to aryl ethers 6 under Mitsunobu conditions, with a broad range of substituted phenols. Alternatively, quantitative conversion of hydroxyl to bromide followed by displacement with alkyl and aryl thiols gave thioethers 8. Thioethers were optionally oxidized to sulfones 9. Bromide displacement by azide, followed by reduction to amine and acylation with a range of carboxylic acids and sulfonyl chlorides gave carboxamides 11 and sulfonamides 13, respectively. Crude purity at typically >90% was observed for each of the five modifications detailed. A series of 20 compounds, exemplifying each modification, was reprepared, purified, and fully characterized. PMID- 11029175 TI - An improved procedure for N- to C-directed (Inverse) solid-phase peptide synthesis. AB - A method for solid-phase peptide synthesis in the N- to C-direction that delivers good coupling yields and a low degree of epimerization is reported. The optimized method involves the coupling, without preactivation, of the resin-bound C terminal amino acid with excess amounts of amino acid tri-tert-butoxysilyl (Sil) esters, using HATU as coupling reagent and 2,4,6-trimethylpyridine (TMP, collidine) as a base. For the amino acids investigated, the degree of epimerization was typically 5%, except for Ser(t-Bu) which was more easily epimerized (ca. 20%). Five tripeptides (AA(1)-AA(2)-AA(3)) with different properties were used as representative model peptides in the development of the synthetic method: Asp-Leu-Glu, Leu-Ala-Phe, Glu-Asp-Val, Asp-Ser-Ile, and Asp-D Glu-Leu. The study used different combinations of HATU and TBTU as activating agents, N, N-diisopropylethylamine (DIEA) and TMP as bases, DMF and dichloromethane as solvents, and cupric chloride as an epimerization suppressant. The epimerization of AA(2) in the coupling of AA(3) was further reduced in the presence of cupric chloride. However, the use of this reagent also resulted in a decrease in loading onto the resin and significant cleavage between AA(1) and AA(2). Experiments indicated that the observed suppressing effect of cupric chloride on epimerization in the present system merely seemed to be a result of a base-induced cleavage of the oxazolone system, the key intermediate in the epimerization process. Consequently, the cleavages were most pronounced in slow couplings. An improved synthesis of fully characterized amino acid tri-tert butoxysilyl (Sil) ester hydrochloride building blocks is presented. The amino acid Sil esters were found to be stable as hydrochlorides but not as free bases. Although only a few peptides have been used in this study, we believe that the facile procedure devised herein should provide an attractive alternative for the solid-phase synthesis of short (six residues or less) C-terminally modified peptides, e.g., in library format. PMID- 11029176 TI - Solid phase synthesis of tertiary amines on amide REM resins: Grignard and metal hydride compatible resins. AB - Four new amide REM resins (AM REM 2-5) are described, and their use is illustrated for the synthesis of tertiary amines 6-9 and 13-16. Amide REM resins 4 and 5, which have a phenyl ring attached to the amide nitrogen, are found to give superior product yields and purities, and the resins are stable to a wider range of reagents and conditions compared to REM resin 1. PMID- 11029177 TI - Synthesis of a 10,000 member 1,5-benzodiazepine-2-one library by the directed sorting method. AB - The solid-phase synthesis of a 10,000 member combinatorial library of 1,5 benzodiazepine-2-one derivatives is reported. The 3-amino-1,5-benzodiazepine-2 one scaffold was prepared in solution, and the benzamide nitrogen was used as a point of attachment to the resin. The 5-aniline and 3-amine were then used as points of diversity. A 10,000 member library was synthesized using the Irori directed sorting system, and after analysis of a representative sample from the library, the Irori system was used to remove the compounds of lower purity. PMID- 11029178 TI - Solid-phase synthesis of a farnesylated CaaX peptide library: inhibitors of the Ras CaaX endoprotease. AB - A solid-phase method, based on Kaiser's p-benzophenone oxime resin, was developed for the synthesis of a series of N-acetyl-S-(E, E-farnesylated) Ca(1)a(2)X tetrapeptides as potential inhibitors of recombinant Ras and a-factor converting enzyme (RCE). N-Acetyl-S-(E, E-farnesyl)-L-cysteine was coupled to resin-bound a(1)a(2) dipeptide using HOBt/DCC activation in conjunction with N-BOC chemistry. The protected farnesylated tripeptide was cleaved from the resin with simultaneous addition of the X residue by treating the resin-bound farnesylated Ca(1)a(2) tripeptide with L-amino acid benzyl ester tosylates under mildly acidic conditions. The benzyl ester was saponified, and the resulting carboxylate precipitated by ether to afford a library of tetrapeptides as a mixture of diastereomers at the cysteine center. The peptides were evaluated as inhibitors of recombinant yeast RCE endoprotease (yRCE) to obtain information about the affinity of the enzyme for the a(1)a(2)X portion of the Ca(1)a(2)X moiety. PMID- 11029180 TI - Parallel electrosynthesis of alpha-alkoxycarbamates, alpha-alkoxyamides, and alpha-alkoxysulfonamides using the spatially addressable electrolysis platform (SAEP) AB - The spatially addressable electrolysis platform (SAEP) has been designed and constructed. It is demonstrated that the advantages of electrochemistry can be readily adapted to combinatorial chemistry and parallel synthesis formats. Parallel electrosynthesis of alpha-alkoxycarbamates, alpha-alkoxyamides, and alpha-alkoxysulfonamides via anodic oxidation of carbamates, amides, and sulfonamides, respectively, highlights the main features of the SAEP. PMID- 11029179 TI - Polymer-assisted solution-phase synthesis of 2'-amido-2'-deoxyadenosine derivatives targeted at the NAD(+)-binding sites of parasite enzymes. AB - A polymer-assisted solution-phase (PASP) synthesis of lead structure analogues ready for biological testing without the demand for chromatographic purification is described. Carboxylic acids are coupled to the Kenner or Ellman safety catch linker, respectively, activated by methylation or cyanomethylation and subsequently transferred to the 2'-amino group of the 2'-amino-2'-deoxyadenosine scaffold (5). The chemoselective attack of weakly nucleophilic amino groups on the N-alkylated N-acyl sulfonamide linker allows for the synthesis of amides 6 in high yields without the need for protection of primary and secondary hydroxyl functions. Thus, the use of 4-sulfamylbenzoylaminomethyl polystyrene is reported for the construction of chemoselective polymer-supported acylating reagents instead of its known use as linker in solid-phase peptide or organic synthesis. This approach is demonstrated to be well suited to obtain 2'-amido-2' deoxyadenosine derivatives 6 in parallel format. Biological evaluation of all compounds reported revealed no improvement over known lead structures. PMID- 11029181 TI - Rasta silanes: new silyl resins with novel macromolecular architecture via living free radical polymerization AB - Heating TEMPO-methyl resin with dialkylsilane styrenes affords larger resin beads via living free radical polymerization. The new silyl resins prepared by this solvent-free suspension polymerization protocol have been coined "Rasta silanes". Rasta silanes have a novel macromolecular architecture typified by long straight chain polymers bearing the silanes which emanate from the phenyl rings of a cross linked polystyrene core. By careful selection of comonomers during the polymerization step, loading capacity, silane spacing, and the relative distance of the silane moieties from the resin core can be controlled. The consistently high-loading Rasta silane resins produced can be easily converted into either a reactive silyl chloride or triflate to subsequently anchor alcohols and phenols to the solid phase. Cleavage from the resin can be mediated by treatment with HF.pyridine, TFA solutions, or TBAF. PMID- 11029182 TI - Site-selective DNA alkylation of GG steps by naphthaldiimide derivatives possessing enantiomeric epoxide. AB - We have synthesized an enantiomeric pair of novel DNA alkylating agents consisting of a naphthaldiimide intercalator and a chiral epoxy side chain. These naphthaldiimide derivatives have high DNA binding affinity and selectively alkylate 5'G of the GG steps for (S)-epoxide and 3'G for (R)-epoxide. PMID- 11029183 TI - Reaction of 2-bromomethylazoles and TosMIC: A domino process to azolopyrimidines. Synthesis of core tricycle of the variolins alkaloids. AB - A new reaction of N-protected 2-bromomethylazoles and tosylmethyl isocyanide (TosMIC) leading to the preparation of azolopyrimidines is described. This domino sequence was used to synthesize the pyrido[3',2':4,5]pyrrolo[1,2-c]pyrimidine core of alkaloids variolins from 4-methoxy-2-methylpyrrolo[2,3-b]pyrimidine in two steps. PMID- 11029185 TI - Absolute configuration of 1,n-diols by NMR: the importance of the combined anisotropic effects in bis-arylmethoxyacetates AB - The absolute configuration of a 1,n-diol can be assigned from the (1)H NMR spectra of its (R)- and (S)-AMAA diesters if the chemical shifts are interpreted as the result of the joint action of the two chiral auxiliaries. PMID- 11029184 TI - Direct correlation between complex conformation and chiral discrimination upon inclusion of amino acid derivatives by beta- and gamma-cyclodextrins. AB - A correlation between the conformation and chiral recognition characteristics of a series of modified amino acid complexes with beta- and gamma-cyclodextrins has been determined, using titration microcalorimetry and (1)H NMR techniques. The enantiomeric discrimination (D or L) is found to be dependent on the adoption of one of two distinct conformations. With the magnitude of the chiral discrimination (K(D)/K(L)) arising from the guest's depth of penetration into the host's cavity. PMID- 11029186 TI - Synthesis and association behavior of butadiyne-bridged AB - Butadiyne-bridged [4(4)](2,6)pyridinophane and [4(6)](2, 6)pyridinophane derivatives were synthesized, and their heteroassociations with the corresponding metacyclophanes and complexations with organic cations were investigated. PMID- 11029187 TI - Synthesis, utility, and X-ray crystal structure of novel complexes of baccatin III with imidazole and 2-propanol. AB - Baccatin III forms crystalline complexes 4 and 5 with imidazole and 2-propanol, respectively. These compounds are useful in the purification of baccatin III from mixtures of taxanes derived from plant-cell fermentation. PMID- 11029188 TI - Two-dimensional molecular layers: interplay of H-bonding and van der waals interactions in the self-assembly of N,N'-dialkylsulfamides AB - N,N'-Dialkylsulfamide molecules assemble into solid-state structures consisting of 2D layers. The 2D layers are based on a hydrogen-bonded network of the sulfamide groups and the close-packing of alkyl groups on both sides of the 2D H bonded network. The thickness of a 2D layer is proportional to the size of the alkyl substituents. The interplay of H-bonding and van der Waals interactions leads to stable 2D layers that pack into 3D structures. PMID- 11029189 TI - A lead-filled G-quadruplex: insight into the G-Quartet's selectivity for Pb(2+) over K(+) AB - The lipophilic nucleoside, G 1, extracts Pb(2+) picrate from water into organic solvents to give structures based on the hydrogen-bonded G-quartet. Crystal structures indicate important differences between (G 1)(8)-Pb(2+) and (G 1)(8) K(+). The divalent Pb(2+) templates a smaller G(8) cage than does K(+), as judged by the M-O6 bond length, O6-O6 diagonal distance, and inter-tetramer separation. The more compact Pb(2+) octamer correlates with NMR data indicating that N2-N7 hydrogen bonds in (G 1)(8)-Pb(2+) are kinetically more stable than in (G 1)(8) K(+). PMID- 11029190 TI - A crystallization-induced stereoselective glycosidation reaction in the synthesis of the anticancer drug etoposide. AB - The anticancer drug etoposide, 1, is prepared in 79% overall yield from readily available 4'-demethyl-4-epipodophyllotoxin, 3, and 4, 6-O-ethylidene-2,3-O dibenzyl-D-glucose, 4, via a crystallization-induced stereoselective glycosidation reaction followed by catalytic hydrogenation. PMID- 11029191 TI - Potassium tert-butoxide catalyzed addition of carbonyl derivatives to styrenes AB - A catalytic amount of t-BuOK in DMSO allows the addition of ketones or imines to styrenes at 40 degrees C in good to excellent yield. Nitriles add to styrenes in NMP or in DMSO at room temperature. PMID- 11029193 TI - Steric and conformational control of the regioselectivities in the ene reaction with trisubstituted cycloalkenes: comparison of the enophiles singlet oxygen, triazolinedione, and nitrosoarene AB - The nitrosoarene ene reaction with the cycloalkenes 1-3 and E-4 proceeds in high twix regioselectivity to afford the hydroxylamine ene products 1a-4a (twix) and 1b-4b (twin, except far E-4 twix). Steric interactions in the enophile attack are responsible for the skew trajectory of the nitrosoarene enophile. For Z-1 methylcyclooctene (Z-4), twin abstraction dominates, caused by conformational constraints (transannular interactions) in the hydrgogen-atom abstraction. The balance between these steric and conformational factors dictates the regioselectivity in the ene reaction PMID- 11029194 TI - Reactions of novel reactive species generated by Samarium(II)-mediated one electron reduction of fischer-type carbene complexes AB - Two types of novel reactive species were generated by one-electron reductions of Fischer-type carbene complexes. In the case of tungsten complexes, one-electron reductions generated anion radical species, which added to electron-deficient olefins to give addition products. In the case of chromium complexes, carbonyl insertion occurred to give acyl chromate complexes, which underwent 1, 4-addition to various electron-deficient olefins. PMID- 11029192 TI - Titanium(IV)-mediated tandem deprotection-cyclodehydration of protected cysteine N-amides: biomimetic syntheses of thiazoline- and thiazole-containing heterocycles. AB - The scope and limitations of TiCl(4)-mediated Delta(2)-thiazoline synthesis via tandem deprotection-dehydrocyclization of trityl-protected cysteine N-amides is presented. While chemical yields are acceptable (53-96%), the stereochemical outcomes vary on the basis of structural considerations and reaction conditions (22-99% ee). Racemization at the C(2)-exomethine position limits the utility of this method for the formation of a thiazoline within a peptide. Treatment of a tritylated Cys-Cys dipeptide with TiCl(4) afforded the corresponding thiazole thiazoline heterocycle 12 (38% yield, 97% ee). PMID- 11029195 TI - Synthesis of a primary metabolite of cannabidiol. AB - Cannabidiol 1 is the major nonpsychotropic, neutral constituent in most cannabis preparations. It is devoid of the psychoactive properties typical of cannabis; however, it produces numerous, potentially therapeutic pharmacological effects, some of which may be due to its metabolites. We report now the first total synthesis of 7-hydroxycannabidiol 2, a primary metabolite of cannabidiol, in an eight-step procedure. PMID- 11029196 TI - Site-selective and stereoselective intramolecular (2pi + 2pi) photocycloaddition of arylalkenes to pyrene and its photocycloreversion AB - Irradiation of a benzene solution containing methyl p-(1 pyrenylmethoxymethyl)cinnamate (1a) with a high-pressure Hg lamp through Pyrex filter stereoselectively gave an intramolecular (2pi + 2pi) photocycloadduct (2a) in an 83% yield in a site-selective manner at the 4,5-position of the pyrene ring. Similar irradiation of an ortho-substituted derivative (3) afforded the corresponding (2pi + 2pi) cycloadduct (4) as a sole product at the 9,10-position of pyrene. The site-selective photocycloaddition can be reasonably explained by the intramolecular sandwich-type singlet exciplexes between the pyrene and phenyl rings. PMID- 11029197 TI - A solid-phase synthesis of N,N'-disubstituted ureas and Perhydroimidazo AB - An efficient method for trapping isocyanates, generated from the Curtius rearrangement, with resin-bound amines is reported. A commercially available carboxylic acid is treated with diphenylphosphoryl azide, followed by thermal rearrangement, cooling, and trapping in one pot. Cleavage from the resin gives an N, N'-disubstituted urea in excellent purity, as demonstrated with several heterocyclic and aliphatic carboxylic acids. Further utility is shown by preparing several novel perhydroimidazo[1,5-a]pyrazines. PMID- 11029198 TI - Conformational study of the intramolecular diels-alder reaction of a pentadienyl acrylate. Theoretical evaluation of kinetic and thermodynamic control AB - Acrylate 4, prepared from diacetylrhamnal, underwent intramolecular Diels-Alder cycloaddition to give the thermodynamically disfavored trans-fused gamma-lactone 15 as the major product, along with two stereoisomeric cycloadducts. A computational analysis of each of the four transition states arising from 4 and the corresponding cycloadducts permits an understanding of the contrasting requirements for kinetic versus thermodynamic control of the reaction. PMID- 11029199 TI - Increasing the efficiency of the transannular diels-alder strategy via stille macrocyclizations AB - To increase the potential and flexibility of the transannular Diels-Alder strategy to build tricycles and tetracycles, the synthesis of macrocyclic trienes of defined geometries has been approached via Stille macrocyclization, giving very high yield and purity of the desired macrocycles or tricycles. PMID- 11029200 TI - Hetero Diels-Alder reactions of 1-amino-3-siloxy-1,3-butadienes under strictly thermal conditions. AB - The hetero Diels-Alder reaction of 1-amino-3-siloxy-1,3-butadiene (1a) with a range of unactivated aldehydes proceeds readily under remarkably mild conditions: at room temperature and in the absence of Lewis acid catalysts. The cycloadducts are formed in good yields and can be converted directly to the corresponding dihydro-4-pyrones using acetyl chloride. Ketones and imines are also reactive in hetero Diels-Alder reactions with this diene. PMID- 11029201 TI - Synthesis of 2-alkylidenecyclopentanones via palladium-catalyzed cross-coupling of 1-(1-Alkynyl)cyclobutanols and aryl or vinylic halides AB - The palladium-catalyzed cross-coupling of aryl or vinylic halides and 1-(1 alkynyl)cyclobutanols affords good yields of stereoisomerically pure 2-arylidene- or 2-(2-alkenylidene)cyclopentanones, respectively, by a process involving (1) oxidative addition of the organic iodide to Pd(0), (2) carbopalladation of the triple bond of the 1-(1-alkynyl)cyclobutanol, (3) regio- and stereoselective ring expansion to form a novel palladiacycle, and (4) reductive elimination to the 2 alkylidenecyclopentanone with simultaneous regeneration of the Pd(0) catalyst. PMID- 11029202 TI - Synthesis of 9-alkylidene-9H-fluorenes by a novel palladium-catalyzed rearrangement AB - In the presence of a palladium catalyst and NaOAc, aryl iodides react with 1-aryl 1-alkynes to afford 9-alkylidene-9H-fluorenes in good yields. This process appears to involve (1) oxidative addition of the aryl iodide to Pd(0), (2) alkyne insertion, (3) rearrangement of the resulting vinylic palladium intermediate to an arylpalladium species, and (4) aryl-aryl coupling with simultaneous regeneration of the Pd(0) catalyst. PMID- 11029203 TI - Regioselective synthesis of medium-sized bicyclic butenolides by lewis acid catalyzed cyclization of cyclic 1, 3-Bis(trimethylsilyloxy)-1,3-butadienes with oxalyl chloride AB - A new strategy for the synthesis of medium-sized bicyclic gamma alkylidenebutenolides is reported which involves Me(3)SiOTf-catalyzed cyclization of cyclic 1, 3-bis(trimethylsilyloxy)-1,3-butadienes with oxalyl chloride. PMID- 11029205 TI - Orthosilicate-mediated esterification of monosubstituted phosphinic acids AB - Monosubstituted phosphinic acids are esterified with orthosilicates in excellent yields. Phosphinylidene-containing acids react selectively under these conditions, while disubstituted phosphinic acids and phosphonic acids remain unchanged. One-pot procedures are also described for the preparation of phosphinate esters from an alcohol. This novel method provides a convenient and general alternative to more commonly employed conditions such as diazomethane or carbodiimide. PMID- 11029204 TI - Synthesis of C-protected alpha-amino aldehydes of high enantiomeric excess from highly epimerizable N-protected alpha-amino aldehydes. AB - A new procedure for the preparation of C-protected alpha-amino aldehydes of high enantiomeric excess is illustrated using five differently substituted alpha-(N Fmoc)amino aldehydes as starting materials. Highly epimerization-prone substrates were converted to the corresponding morpholino nitrile-protected alpha-amino aldehydes with minimal racemization (products >/= 89% ee). Morpholino nitrile derivatives of phenylglycinal were crystallized and subjected to X-ray structural analysis, allowing for definitive determination of the stereochemistry of amino nitrile formation. A rationale for the stereoselectivity of amino nitrile formation is presented. PMID- 11029206 TI - Diels-alder reactions of hexafluoro-2-butyne with 2-heterosubstituted furans: A facile and general synthesis of 1, 4-disubstituted 2,3 Di(trifluoromethyl)benzenes AB - An electron-donating heteroatom substituent at position-2 of a furan promotes regiospecific opening of the 7-oxa bridge of the Diels-Alder cycloadduct with hexafluoro-2-butyne, producing a 4-heterosubstituted 2,3 di(trifluoromethyl)phenol building block in a single step. The phenol and heteroatom substituent are easily transformed to the corresponding iodide or triflate that readily undergoes Heck, Suzuki, and Stille reactions to install a variety of substituents in high yields. This methodology provides a facile and general synthesis of 1,4-disubsituted 2, 3-di(trifluoromethyl)benzenes. PMID- 11029207 TI - Solid-phase synthesis of bis-2-imidazolidinethiones from resin-bound tripeptides. AB - The exhaustive reduction of resin-bound tripeptides with borane afforded three secondary amines and one primary amine. The treatment of the solid-support polyamines with thiocarbonyldiimidazole afforded, following cleavage of the solid support, the corresponding bis-cyclic thiourea in good purity and yield. PMID- 11029208 TI - Using symmetry to monitor geared bond rotation in aromatic amides by dynamic NMR AB - Dynamic NMR proves that the fastest interconversion between conformers of simple tertiary aromatic amide 3 is racemization via a geared (correlated) rotation of both the Ar-CO and N-CO bonds. The symmetry of 3 is such that correlated and noncorrelated rotations are easily distinguishable, even without assignment of the NMR spectrum, simply by observing the number of peaks taking part in the exchange. PMID- 11029209 TI - Free radical allyl transfers utilizing soluble non-cross-linked polystyrene and carbohydrate scaffold supports AB - Free radical allylations were studied using (1) soluble non-cross-linked polystyrene supports, (2) carbohydrate scaffolds, and (3) a combination of both synthetic motifs. Allylations on these custom designer supports provide easily purified products, free of tin residues. A D-xylose carbohydrate scaffold bearing a bromoester was used for a diastereoselective allyl tin transfer thermally at 80 degrees C and with Lewis acids. This is the first example of a diastereoselective radical reaction directed by a removable polymer-supported carbohydrate auxiliary. PMID- 11029210 TI - Rearrangement of phenylethenes on reaction with iodine-xenon difluoride AB - Phenyl-substituted ethenes react with iodine and xenon difluoride to provide difluorinated products. Iodofluoro intermediates react with xenon difluoride to produce transient iodine difluoride species that lose IF and F(-) and produce carbocations. PMID- 11029211 TI - Stereoselective synthesis of the alpha-glycoside of a KDO "C"-disaccharide. AB - The reaction of tert-butyl (4,5,7, 8-tetra-O-acetyl-3-deoxy-alpha-D-manno-2 octulopyranosyl chloride)onate donor 7 with the 6-formylgalactopyranoside acceptor 4 in the presence of SmI(2) provided only the KDO alpha-C-disaccharide 8. The bulky tert-butyl ester in the donor was used to reverse the stereochemical outcome of C-glycosylation, stereoselectively forming the alpha-"C"-disaccharide of KDO. PMID- 11029212 TI - Total synthesis of (+)-crocacin C. AB - The first asymmetric synthesis of (+)-crocacin C (3) is described which served to confirm the absolute configuration of this compound. The key step in the sequence was the stereoselective assembly of the (E,E)-diene amide side chain by a Stille cross-coupling between the stannane 5 and vinyl iodide 6. PMID- 11029213 TI - Laser flash photolysis studies of alkoxyl radical kinetics using 4 nitrobenzenesulfenate esters as radical precursors AB - 4-Nitrobenzenesulfenate esters were used as precursors for the generation of alkoxyl radicals under laser flash photolysis conditions. The esters were efficiently cleaved using the Nd:YAG third harmonic (355 nm) to produce alkoxyl radicals and the 4-nitrobenzenethiyl radical. Rate constants for beta-scission and 1, 5-hydrogen abstraction reactions of alkoxyl radicals were measured. PMID- 11029214 TI - Stille-type cross-coupling-An efficient way to various symmetrically and unsymmetrically substituted methyl-bipyridines: toward new ATRP catalysts AB - Various mono- and disubstituted 2,2'-bipyridines were synthesized in high yields and multigram scales utilizing Stille-type coupling procedures. The corresponding bromo-picoline and tributyltin-picoline building blocks were prepared from commercially available amino-picoline compounds. As first examples of metal complexes, 4,5'-dimethyl-2,2'-bipyridine was reacted with copper(II) and iron(II) ions and investigated as catalyst in ATRP. PMID- 11029215 TI - Ozonolysis in the presence of Lewis acids: directed addition to carbonyl oxides. AB - The presence of Lewis acids strongly influences the distribution of products from alkene ozonolysis. Delivery of alkoxide and phenoxide ligands to a coordinated carbonyl oxide affords hydroperoxyacetals under aprotic conditions. PMID- 11029216 TI - Norrish type II photochemical reaction of an aryl ketone on a monolayer-protected gold nanocluster. Development Of a probe of conformational mobility AB - A Norrish type II reaction of an aryl ketone monolayer-protected colloid was investigated for the first time. Photolysis of mercaptoundecanophenone-modified gold colloid, MUP-MPC, generates free acetophenone in solution and the nonene modified MPC via the triplet excited state and the 1,4-biradical intermediate. The reaction is being developed as a probe of conformational mobility in these unique environments. PMID- 11029217 TI - Hydrogen iodide-promoted reduction of beta-chlorovinamidinium salts AB - HI cleanly reduced chlorovinamidinium salts to the vinamidinium salt in essentially quantitative assay yield and 55-85% isolated yield following recrystallization. The reaction proceeds via protonation of the beta-carbon atom of the vinamidinium and dechlorination via the formation of I-Cl. PMID- 11029218 TI - Chemoselective reactions of amidines: selective formation of iminopyrimidine regioisomers AB - The dramatic effect of base on the chemoselectivity of the reaction of amidines with substituted 3-phenyl-2-propynylnitriles is demonstrated. Amidine 1 can be added to cyanoalkyne 2 to give iminopyrimidine isomer 3 with high selectivity. The addition of 2 equiv of NaHMDS completely reverses the selectivity of the reaction, yielding isomer 4 almost exclusively. This method has been used to prepare a variety of substituted 4-iminopyrimidines. PMID- 11029219 TI - N-Benzylhydroxylamine addition to beta-aryl enoates. Enantioselective synthesis of beta-aryl-beta-amino acid precursors AB - Chiral Lewis acid catalyzed N-benzylhydroxylamine addition to pyrrolidinone derived enoates afforded beta-aryl-beta-amino acid derivatives in high enantiomeric purity with moderate to very good chemical efficiency. PMID- 11029221 TI - Perfect switching of photoreactivity by acid: photochemical decarboxylation versus transesterification of mesityl cyclohexanecarboxylate AB - Mesityl cyclohexanecarboxylate has been photodecarboxylated upon excitation at 254 nm to form cyclohexylmesitylene in good yield in neutral acetonitrile solutions. In the presence of a catalytic amount of the acid and ethanol as a nucleophile, however, the same compound undergoes facile transesterification upon irradiation, affording its ethyl ester in almost quantitative yield. PMID- 11029220 TI - Solid-phase synthesis of bleomycin group antibiotics. Elaboration of deglycobleomycin A(5). AB - The solid-phase syntheses of two deglycobleomycin A(5) analogues were achieved using a commercially available polystyrene resin containing triphenylmethyl linked spermidine. The final products were deblocked and released from the resin, analyzed, and purified by C(18) reversed phase HPLC and characterized by high field (1)H NMR spectroscopy and mass spectrometry. The purified products relaxed supercoiled plasmid DNA in a concentration-dependent fashion and to the same extent as authentic material derived from natural BLM A(5). PMID- 11029222 TI - Oxidation of electron-deficient anilines by HOF. A route to nitro-containing compounds for molecular electronic devices AB - Nitroaromatic compounds for molecular electronic devices are prepared by the high yielding oxidation of electron-deficient anilines using HOF generated in a fluorine-acetonitrile-water system. PMID- 11029224 TI - Synthesis of glycosyl-1-phosphates via dehydrative glycosylation PMID- 11029223 TI - Total synthesis of (-)-ascochlorin via a cyclobutenone-based benzannulation strategy. AB - The application of a convergent benzannulation strategy in an efficient synthesis of (-)-ascochlorin is described. PMID- 11029225 TI - The post concussional syndrome after mild head injury: some practical considerations. PMID- 11029226 TI - Low grade astrocytomas: controversies in management. AB - Low grade astrocytomas are common brain neoplasms that primarily affect young adults. Although these patients often have a reasonably long survival, most will ultimately succumb to their tumours. Often the tumours progress to higher grade astrocytomas. The optimal management plan for these tumours is controversial and ranges from observation to macroscopic excision, radiotherapy and chemotherapy. The evidence for each of these approaches is presented in this review and a management algorithm is presented. PMID- 11029227 TI - Treatment of Parkinson's disease. AB - The aim of current treatment of Parkinson's disease is to ameliorate the symptoms while seeking to lessen the potential development of late levodopa complications. To this end, there is ample evidence that the early use of dopamine agonists is beneficial in younger Parkinsonian patients but monotherapy with dopamine agonists is for only a select few. Nonergot dopamine agonists offer the potential for fewer side effects. Lower dose levodopa therapy delays the onset and reduces severity of dyskinesia and end of dose failure. However levodopa remains the treatment of choice in Parkinson's disease and should not be restricted unnecessarily in patients with disability. There is no evidence that levodopa is toxic to dopaminergic neurons in people with Parkinson's disease. As yet, no drugs are of proven neuroprotective value. Dopamine agonists, catechol-o methyltransferase inhibitors, amantadine and apomorphine have differing but beneficial roles in the management of levodopa side effects. Ablative surgery and deep brain stimulation of thalamus, globus pallidus and subthalamic nucleus are increasingly available but choice of procedure depends not just on patient symptomatology, but also on local experience and results. Ideally, deep brain stimulation is the treatment of choice as it offers less morbidity than bilateral ablative surgery, the possibility of postoperative adjustments and the potential for reversibility if better treatments become available. PMID- 11029229 TI - Was Hippocrates a beginner at trepanning and where did he learn? AB - A critical reading of Hippocrates'On Wounds of the Head suggests the writer was a beginner at trepanning. He could not explain clearly, in the pathological terms used in his time, why routine early trepanning was beneficial. His technique was hesitant, suggesting he himself had never done the routine early trepanning he advocated, and he had not yet realised that his policy would be unacceptable to most Greek patients. It is suggested that he might have learnt trepanning on a brief trip to Marseilles, where the Gauls had already trepanned for 1500 years. PMID- 11029228 TI - The postconcussion syndrome after mild head trauma part II: is migraine underdiagnosed? AB - The evidence for post-traumatic migraine as the cause of the postconcussion syndrome in a proportion of patients is reviewed. CONCLUSION: patients suffering recurrent post-traumatic headaches or other elements of the postconcussion syndrome should be treated for migraine. PMID- 11029230 TI - Risk factors for poor outcome of untreated arteriovenous malformation. AB - This study was conducted to determine risk factors for poor outcome in the natural history of arteriovenous malformation (AVM). We statistically analysed the correlation between clinical or angiographical findings and clinical outcomes for 55 cases of untreated AVM. Subsequent haemorrhage from AVMs was the only significant risk factor for poor outcome (P< 0.0001). The odds ratio was 44.56 with a 95% confidence interval (CI) from 4.80 to 413.90. Risk factors for subsequent haemorrhage from AVMs were also determined. The size (P = 0.0483) and location (P = 0.0147) of an AVM were significant risk factors for subsequent haemorrhage. The odds ratios were 3.97 with a 95% CI from 1.18 to 13.33 and 3.89 with a 95% CI from 1.10 to 13.72, respectively. AVMs of more than 60 mm, and deep or infratentorial AVMs had more chance of subsequent haemorrhage, and hence of a poor outcome. We propose using an aggressive multidisciplinary approach to treating these AVMs. PMID- 11029231 TI - The preventability of death in road traffic fatalities with head injury in Victoria, Australia. The Consultative Committee on Road Traffic Fatalities. AB - Since 1992 the Consultative Committee on Road Traffic Fatalities in Victoria (CCRTF) has examined the medical management of patients who died following motor vehicle accidents. Three hundred and fifty-five fatalities with head injury occurring between 1 July, 1992 and 31 December 1997 were assessed by the CCRTF. They represented 79% of the total 449 fatalities examined by the Committee. Following examination of the complete medical records and multidisciplinary discussion, the Committee considered 237 (67%) of the 355 neurotrauma deaths to be non-preventable, 105 (30%) potentially preventable and 13 (4%) preventable. The present analysis excludes the non-preventable deaths in order to focus on preventable factors. Problems identified in the 118 patients pre-hospital included: no intubation; prolonged scene time; and no intravenous access; in 139 emergency room attendances: inappropriate reception including delay in arrival of a consultant, no neurosurgical consultation, no CT scan of the head, inadequate blood gases and oxygen monitoring, inadequate fluid resuscitation, delayed respiratory resuscitation and delayed dispatch to the operating room; in 111 operating room visits: no ICP monitoring, inadequate fluid administration and inappropriate anaesthetic technique; and in 90 intensive care unit admissions: no ICP monitoring. Overall, 1745 individual problems in the various areas of care were identified, of which 1104 (63%) were judged to have contributed to death. Improved delivery and quality of trauma care could reduce the identified problems in emergency services and clinical management. Basic principles of trauma management remain the most important means of reducing morbidity and death following road trauma. The leadership role of the neurosurgeon in neurotrauma care is emphasised. PMID- 11029232 TI - Dissecting aneurysms of the posterior inferior cerebellar artery: report of four cases and review of the literature. AB - Dissecting aneurysms frequently involve the vertebral arteries and their branches, but those exclusively on the posterior inferior cerebellar artery (PICA) represent only 24 cases in the literature, including the four cases discussed in this article. The clinical diagnosis lacks pathognomonic signs or symptoms, with presentations such as subarachnoid haemorrhage or ischaemia of the brain stem or the cerebellum, and the management is controversial. Wrapping, clipping and embolisation of the aneurysms were tried in this series with different outcomes. Exclusion of the pathological segment should be performed, as shown by rebleeding from our case which was wrapped or by progression of the vascular disease in cases where treatment was delayed. Surgical or endovascular occlusion are well tolerated in our cases and in those reported from the literature, which implies the absence of normal perforating branches to the brain stem arising from the proximal dissected segment of the PICA and a good collateral circulation. A revascularisation procedure using the occipital artery can be performed in order to prevent infarction if an endovascular test occlusion is not tolerated. PMID- 11029233 TI - Current management of acoustic neuromas: review of surgical approaches and outcomes. AB - The management options for patients with acoustic neuromas is discussed with a review of 164 patients assessed and treated between 1994 and 1998. Twenty-one patients have neurofibromatosis type II. In 33 cases initial observation was undertaken with repeated imaging. Surgical removal of 122 tumours was performed in 121 patients. Eleven of these patients have NF2, of whom three underwent Auditory Brainstem Implantation. Hearing preservation tumour removal was attempted in 37 and was successful in 20 (54%). The middle cranial fossa approach was used in ten cases with 100% successful hearing preservation. The retrosigmoid approach was used in 27 cases with 36% successful hearing preservation. Non hearing preservation tumour removal was performed in 85 cases where hearing was poor or the tumour measured more than 2 cm within the cerebellopontine angle. The translabyrinthine approach was used in 80 of these patients. Postoperative facial nerve outcome was dependent on tumour size. All 38 patients with tumours 90%). That (86)Rb(+) uptake was almost completely abolished by Sch-28080 indicates an effect of this agent on the Na(+)-K(+) ATPase. ATPase assays in membranes, or lysed cells, demonstrated sensitivity to ouabain but not Sch-28080. Thus the inhibitory effect of Sch-28080 was dependent on cell integrity. (86)Rb(+)-uptake studies without bumetanide demonstrated that ouabain inhibited activity by only 50%. Addition of Sch-28080 (200 microM) blocked all residual activity. Intracellular ATP declined after Sch-28080 (200 microM) but recovered after removal of this agent. In conclusion, high concentrations of Sch-28080 inhibit K(+)-ATPase activity in mouse IMCD-3 (mIMCD 3) cells as a result of ATP depletion. PMID- 11029279 TI - Ca(2+)-activated Cl(-) current in sheep lymphatic smooth muscle. AB - Freshly dispersed sheep mesenteric lymphatic smooth muscle cells were studied at 37 degrees C using the perforated patch-clamp technique with Cs(+)- and K(+) filled pipettes. Depolarizing steps evoked currents that consisted of L-type Ca(2+) [I(Ca(L))] current and a slowly developing current. The slow current reversed at 1 +/- 1.5 mV with symmetrical Cl(-) concentrations compared with 23.2 +/- 1.2 mV (n = 5) and -34.3 +/- 3.5 mV (n = 4) when external Cl(-) was substituted with either glutamate (86 mM) or I(-) (125 mM). Nifedipine (1 microM) blocked and BAY K 8644 enhanced I(Ca(L)), the slow-developing sustained current, and the tail current. The Cl(-) channel blocker anthracene-9-carboxylic acid (9 AC) reduced only the slowly developing inward and tail currents. Application of caffeine (10 mM) to voltage-clamped cells evoked currents that reversed close to the Cl(-) equilibrium potential and were sensitive to 9-AC. Small spontaneous transient depolarizations and larger action potentials were observed in current clamp, and these were blocked by 9-AC. Evoked action potentials were triphasic and had a prominent plateau phase that was selectively blocked by 9-AC. Similarly, fluid output was reduced by 9-AC in doubly cannulated segments of spontaneously pumping sheep lymphatics, suggesting that the Ca(2+)-activated Cl( ) current plays an important role in the electrical activity underlying spontaneous activity in this tissue. PMID- 11029280 TI - Voltage dependence of L-arginine transport by hCAT-2A and hCAT-2B expressed in oocytes from Xenopus laevis. AB - Membrane potential and currents were investigated with the two-electrode voltage clamp technique in Xenopus laevis oocytes expressing hCAT-2A or hCAT-2B, the splice variants of the human cationic amino acid transporter hCAT-2. Both hCAT-2A and hCAT-2B-expressing oocytes exhibited a negative extracellular L-arginine concentration ([L-Arg](o))-sensitive membrane potential, additive to the K(+) diffusion potential, when cells were incubated in Leibovitz medium (containing 1.45 mM L-Arg and 0.25 mM L-lysine). The two carrier proteins produced inward and outward currents, which were dependent on the L-Arg gradient and membrane potential. Ion substitution experiments showed that the hCAT-induced currents were independent of external Na(+), K(+), Ca(2+), or Mg(2+). The apparent Michaelis-Menten constant values at -60 mV, obtained from plots of L-Arg-induced currents against [L-Arg](o), were 0.97 and 0.13 mM in oocytes expressing hCAT-2A and hCAT-2B, respectively; maximal currents amounted to -194 +/- 8 and -84 +/- 2 nA, respectively. At saturating [L-Arg](o), the current-voltage relationships of hCAT-2A-expressing oocytes became steeper, yielding an additional conductance up to 2 microS/oocyte, whereas those of hCAT-2B-expressing oocytes were simply shifted to the right, resulting in voltage-independent difference currents. The distinct electrochemical properties of the two isoforms of hCAT-2 are assumed to contribute differentially to the membrane transport and the maintenance of cationic amino acids in various tissues. PMID- 11029281 TI - Substrate flexibility regulates growth and apoptosis of normal but not transformed cells. AB - One of the hallmarks of oncogenic transformation is anchorage-independent growth (27). Here we demonstrate that responses to substrate rigidity play a major role in distinguishing the growth behavior of normal cells from that of transformed cells. We cultured normal or H-ras-transformed NIH 3T3 cells on flexible collagen coated polyacrylamide substrates with similar chemical properties but different rigidity. Compared with cells cultured on stiff substrates, nontransformed cells on flexible substrates showed a decrease in the rate of DNA synthesis and an increase in the rate of apoptosis. These responses on flexible substrates are coupled to decreases in cell spreading area and traction forces. In contrast, transformed cells maintained their growth and apoptotic characteristics regardless of substrate flexibility. The responses in cell spreading area and traction forces to substrate flexibility were similarly diminished. Our results suggest that normal cells are capable of probing substrate rigidity and that proper mechanical feedback is required for regulating cell shape, cell growth, and survival. The loss of this response can explain the unregulated growth of transformed cells. PMID- 11029282 TI - A K(ATP) channel deficiency affects resting tension, not contractile force, during fatigue in skeletal muscle. AB - The objective of this study was to determine how an ATP-sensitive K(+) (K(ATP)) channel deficiency affects the contractile and fatigue characteristics of extensor digitorum longus (EDL) and soleus muscle of 2- to 3-mo-old and 1-yr-old mice. K(ATP) channel-deficient mice were obtained by disrupting the Kir6.2 gene that encodes for the protein forming the pore of the channel. At 2-3 mo of age, the force-frequency curve, the twitch, and the tetanic force of EDL and soleus muscle of K(ATP) channel-deficient mice were not significantly different from those in wild-type mice. However, the tetanic force and maximum rate of force development decreased with aging to a greater extent in EDL and soleus muscle of K(ATP) channel-deficient mice (24-40%) than in muscle of wild-type mice (7-17%). During fatigue, the K(ATP) channel deficiency had no effect on the decrease in tetanic force in EDL and soleus muscle, whereas it caused a significantly greater increase in resting tension when compared with muscle of wild-type mice. The recovery of tetanic force after fatigue was not affected by the deficiency in 2- to 3-mo-old mice, whereas in 1-yr-old mice, force recovery was significantly less in muscle of K(ATP) channel-deficient than wild-type mice. It is suggested that the major function of the K(ATP) channel during fatigue is to reduce the development of a resting tension and not to contribute to the decrease in force. It is also suggested that the K(ATP) channel plays an important role in protecting muscle function in older mice. PMID- 11029283 TI - Overexpression of fructose 2,6-bisphosphatase decreases glycolysis and delays cell cycle progression. AB - The ability to overexpress 6-phosphofructo-2-kinase/fructose 2, 6-bisphosphatase (PFK-2)/(FBPase-2) or a truncated form of the enzyme with only the bisphosphatase domain allowed us to analyze the relative role of the kinase and the bisphosphatase activities in regulating fructose 2,6-bisphosphate (Fru-2,6-P(2)) concentration and to elucidate their differential metabolic impact in epithelial Mv1Lu cells. The effect of overexpressing PFK-2/FBPase-2 resulted in a small increase in the kinase activity and in the activity ratio of the bifunctional enzyme, increasing Fru-2,6-P(2) levels, but these changes had no major effects on cell metabolism. In contrast, expression of the bisphosphatase domain increased the bisphosphatase activity, producing a significant decrease in Fru-2,6-P(2) concentration. The fall in the bisphosphorylated metabolite correlated with a decrease in lactate production and ATP concentration, as well as a delay in cell cycle. These results provide support for Fru-2,6-P(2) as a regulator of glycolytic flux and point out the role of glycolysis in cell cycle progression. PMID- 11029284 TI - H(2)O(2) and ethanol act synergistically to gate ryanodine receptor/calcium release channel. AB - We examined the effect of low concentrations of H(2)O(2) on the Ca(2+)-release channel/ryanodine receptor (RyR) to determine if H(2)O(2) plays a physiological role in skeletal muscle function. Sarcoplasmic reticulum vesicles from frog skeletal muscle and type 1 RyRs (RyR1) purified from rabbit skeletal muscle were incorporated into lipid bilayers. Channel activity of the frog RyR was not affected by application of 4.4 mM (0.02%) ethanol. Open probability (P(o)) of such ethanol-treated RyR channels was markedly increased on subsequent addition of 10 microM H(2)O(2). Increase of H(2)O(2) to 100 microM caused a further increase in channel activity. Application of 4.4 mM ethanol to 10 microM H(2)O(2) treated RyRs activated channel activity. Exposure to 10 or 100 microM H(2)O(2) alone, however, failed to increase P(o). Synergistic action of ethanol and H(2)O(2) was also observed on the purified RyR1 channel, which was free from FK506 binding protein (FKBP12). H(2)O(2) at 100-500 microM had no effect on purified channel activity. Application of FKBP12 to the purified RyR1 drastically decreased channel activity but did not alter the effects of ethanol and H(2)O(2). These results suggest that H(2)O(2) may play a pathophysiological, but probably not a physiological, role by directly acting on skeletal muscle RyRs in the presence of ethanol. PMID- 11029285 TI - Mechanical signals and mechanosensitive modulation of intracellular [Ca(2+)] in smooth muscle. AB - We tested the hypothesis that strain is the primary mechanical signal in the mechanosensitive modulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) in airway smooth muscle. We found that [Ca(2+)](i) was significantly correlated with muscle length during isotonic shortening against 20% isometric force (F(iso)). When the isotonic load was changed to 50% F(iso), data points from the 20 and 50% F(iso) experiments overlapped in the length-[Ca(2+)](i) relationship. Similarly, data points from the 80% F(iso) experiments clustered near those from the 50% F(iso) experiments. Therefore, despite 2.5- and 4-fold differences in external load, [Ca(2+)](i) did not deviate much from the length-[Ca(2+)](i) relation that fitted the 20% F(iso) data. Maximal inhibition of sarcoplasmic reticular (SR) Ca(2+) uptake by 10 microM cyclopiazonic acid (CPA) did not significantly change [Ca(2+)](i) in carbachol-induced isometric contractions and isotonic shortening. CPA also did not significantly change myosin light-chain phosphorylation or force redevelopment when carbachol-activated muscle strips were quickly released from optimal length (L(o)) to 0.5 L(o). These results are consistent with the hypothesis and suggest that SR Ca(2+) uptake is not the underlying mechanism. PMID- 11029286 TI - Effect of HCO(3)(-) on TPA- and IBMX-induced anion conductances in Necturus gallbladder epithelial cells. AB - Effects of HCO(3)(-) on protein kinase C (PKC)- and protein kinase A (PKA) induced anion conductances were investigated in Necturus gallbladder epithelial cells. In HCO(3)(-)-free media, activation of PKC via 12-O-tetradecanoylphorbol 13-acetate (TPA) depolarized apical membrane potential (V(a)) and decreased fractional apical voltage ratio (F(R)). These effects were blocked by mucosal 5 nitro-2-(3-phenylpropylamino) benzoic acid (NPPB), a Cl(-) channel blocker. In HCO(3)(-) media, TPA induced significantly greater changes in V(a) and F(R). These effects were blocked only when NPPB was present in both mucosal and basolateral compartments. The data suggest that TPA activates NPPB-sensitive apical Cl(-) conductance (g(Cl)(a)) in the absence of HCO(3)(-); in its presence, TPA stimulated both NPPB-sensitive g(Cl)(a) and basolateral Cl(-) conductance (g(Cl)(b)). Activation of PKA via 3-isobutyl-1-methylxanthine (IBMX) also decreased V(a) and F(R); however, these changes were not affected by external HCO(3)(-). We conclude that HCO(3)(-) modulates the effects of PKC on g(Cl)(b). In HCO(3)(-) medium, TPA and IBMX also induced an initial transient hyperpolarization and increase in intracellular pH. Because these changes were independent of mucosal Na(+) and Cl(-), it is suggested that TPA and IBMX induce a transient increase in apical HCO(3)(-) conductance. PMID- 11029287 TI - Tom20-mediated mitochondrial protein import in muscle cells during differentiation. AB - Mitochondrial biogenesis is accompanied by an increased expression of components of the protein import machinery, as well as increased import of proteins destined for the matrix. We evaluated the role of the outer membrane receptor Tom20 by varying its expression and measuring changes in the import of malate dehydrogenase (MDH) in differentiating C2C12 muscle cells. Cells transfected with Tom20 had levels that were twofold higher than in control cells. Labeling of cells followed by immunoprecipitation of MDH revealed equivalent increases in MDH import. This parallelism between import rate and Tom20 levels was also evident as a result of thyroid hormone treatment. Using antisense oligodeoxynucleotides, we inhibited Tom20 expression by 40%, resulting in 40-60% reductions in MDH import. In vitro assays also revealed that import into the matrix was more sensitive to Tom20 inhibition than import into the outer membrane. These data indicate a close relationship between induced changes in Tom20 and the import of a matrix protein, suggesting that Tom20 is involved in determining the kinetics of import. However, this relationship was dissociated during normal differentiation, since the expression of Tom20 remained relatively constant, whereas imported MDH increased 12-fold. Thus Tom20 is important in determining import during organelle biogenesis, but other mechanisms (e.g., intramitochondrial protein degradation or nuclear transcription) likely also play a role in establishing the final mitochondrial phenotype during normal muscle differentiation. PMID- 11029288 TI - A hyperpolarization- and acid-activated nonselective cation current in Xenopus oocytes. AB - Heterologous expression of a variety of membrane proteins in Xenopus oocytes sometimes results in the appearance of a hyperpolarization-activated inward current. The nature of this current remains incompletely understood. Some investigators have suggested that this current is a Cl current, whereas others have identified it as a nonselective cation current. The purpose of this investigation was to characterize this current in more detail. The hyperpolarization-activated inward current (I(IN)) present in native oocytes was composed of a current carried at least partly by Ca and Mg under physiological ionic conditions plus a Ca-activated Cl current. The Ca-activated Cl current was blocked by chelation of cytosolic Ca with 1,2-bis(2-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid. When Cl currents were blocked, the cation current could be carried by Ca, Mg, or Co, but not appreciably by Ba, Mn, or Cd. I(IN) was stimulated by intracellular acidification. The properties of I(IN) were quite different from those of the store-operated Ca current. Heterologous expression of transient receptor potential-like gene product (TRPL), one of the members of the transient receptor potential family of putative store-operated Ca channels, apparently resulted in alteration of the voltage sensitivity of the endogenous I(IN). PMID- 11029289 TI - The Egr-1 gene is induced by epidermal growth factor in ECV304 cells and primary endothelial cells. AB - The early growth response (Egr)-1 transcription factor serves to couple changes in the extracellular environment to alterations in gene expression. An understanding of the mechanisms that underlie Egr-1 gene regulation should provide important insights into how environmental signals are transduced by endothelial cells. The aim of the present study was to determine whether epidermal growth factor (EGF) induces Egr-1 expression in endothelial cells. In ECV304 cells, Egr-1 mRNA and protein levels were increased in response to EGF. In stable transfection assays, the 1,200-bp promoter of the mouse Egr-1 gene contained information for EGF response via a protein kinase C-independent, mitogen-activated protein kinase-dependent pathway. The endogenous Egr-1 gene was similarly responsive to EGF in primary human umbilical vein endothelial cells, human coronary artery endothelial cells, and rat fat pad endothelial cells, but not in bovine aortic endothelial cells, calf pulmonary artery endothelial cells, or PY-4-1 endothelial cells. Together, these results suggest that the Egr-1 gene is responsive to EGF in a subset of endothelial cells. PMID- 11029290 TI - Molecular characterization of a novel UT-A urea transporter isoform (UT-A5) in testis. AB - Urea movement across plasma membranes is modulated by specialized transporter proteins that are products of two genes, termed UT-A and UT-B. These proteins play key roles in the urinary concentrating mechanism and fluid homeostasis. We have isolated and characterized a 1.4-kb cDNA from testes encoding a new isoform (UT-A5) belonging to the UT-A transporter family. For comparison, we also isolated a 2. 0-kb cDNA from mouse kidney inner medulla encoding the mouse UT-A3 homologue. The UT-A5 cDNA has a putative open reading frame encoding a 323-amino acid protein, making UT-A5 the smallest UT-A family member in terms of molecular size. Its putative topology is of particular interest, because it calls into question earlier models of UT-A transporter structure. Expression of UT-A5 cRNA in Xenopus oocytes mediates phloretin-inhibitable urea uptake and does not translocate water. The distribution of UT-A5 mRNA is restricted to the peritubular myoid cells forming the outermost layer of the seminiferous tubules within the testes and is not detected in kidney. UT-A5 mRNA levels are coordinated with the stage of testes development and increase 15 days postpartum, commensurate with the start of seminiferous tubule fluid movement. PMID- 11029291 TI - Agonist-induced isometric contraction of smooth muscle cell-populated collagen gel fiber. AB - String-shaped reconstituted smooth muscle (SM) fibers were prepared in rectangular wells by thermal gelation of a mixed solution of collagen and cultured SM cells derived from guinea pig stomach. The cells in the fiber exhibited an elongated spindle shape and were aligned along the long axis. The fiber contracted in response to KCl (140 mM), norepinephrine (NE; 10(-7) M), epinephrine (10(-7) M), phenylephrine (10(-6) M), serotonin (10(-6) M), and histamine (10(-5) M), but not acetylcholine (10(-5) M). Phentolamine (10(-7) M) produced a parallel rightward shift of the NE dose-response curve. Moreover, NE induced contraction was partially inhibited by nifedipine and completely abolished by the intracellular Ca(2+) chelator 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester, the myosin light chain kinase inhibitor ML-9, the Rho kinase inhibitor Y-27632, and papaverine. A [(3)H]quinuclidinyl benzilate binding study revealed that the loss of response to acetylcholine was due to the loss of muscarinic receptor expression during culture. The expression of contractile proteins in the fibers was similar to that in cultured SM cells. These results suggest that, although the fiber is not a model for fully differentiated SM, contractile mechanisms are maintained. PMID- 11029292 TI - Hyposmolality stimulates Na(+)/H(+) exchange and HCO(3)(-) absorption in thick ascending limb via PI 3-kinase. AB - The signal transduction mechanisms that mediate osmotic regulation of Na(+)/H(+) exchange are not understood. Recently we demonstrated that hyposmolality increases HCO(3)(-) absorption in the renal medullary thick ascending limb (MTAL) through stimulation of the apical membrane Na(+)/H(+) exchanger NHE3. To investigate the mechanism of this stimulation, MTALs from rats were isolated and perfused in vitro with 25 mM HCO(3)(-)-containing solutions. The phosphatidylinositol 3-kinase (PI 3-K) inhibitors wortmannin (100 nM) and LY 294002 (20 microM) blocked completely the stimulation of HCO(3)(-) absorption by hyposmolality. In tissue strips dissected from the inner stripe of the outer medulla, the region of the kidney highly enriched in MTALs, hyposmolality increased PI 3-K activity 2. 2-fold. Wortmannin blocked the hyposmolality-induced PI 3-K activation. Further studies examined the interaction between hyposmolality and vasopressin, which inhibits HCO(3)(-) absorption in the MTAL via cAMP and often is involved in the development of plasma hyposmolality in clinical disorders. Pretreatment with arginine vasopressin, forskolin, or 8-bromo-cAMP abolished hyposmotic stimulation of HCO(3)(-) absorption, due to an effect of cAMP to inhibit hyposmolality- induced activation of PI 3-K. In contrast to their effects to block stimulation by hyposmolality, PI 3-K inhibitors and vasopressin have no effect on inhibition of apical Na(+)/H(+) exchange (NHE3) and HCO(3)(-) absorption by hyperosmolality. These results indicate that hyposmolality increases NHE3 activity and HCO(3)(-) absorption in the MTAL through activation of a PI 3-K-dependent pathway that is inhibited by vasopressin and cAMP. Hyposmotic stimulation and hyperosmotic inhibition of NHE3 are mediated through different signal transduction mechanisms. PMID- 11029293 TI - Impaired G(s)alpha and adenylyl cyclase cause beta-adrenoceptor desensitization in chronically hypoxic rat hearts. AB - The effects of beta-adrenoceptor stimulation with isoproterenol on electrically induced contraction and intracellular calcium ([Ca(2+)](i)) transient, and cAMP in myocytes from both hypertrophied right and nonhypertrophied left ventricles of rats exposed to 10% oxygen for 4 wk, were significantly attenuated. The increased [Ca(2+)](i) transient in response to cholera toxin was abolished, whereas increased cAMP after NaF significantly attenuated. The biologically active isoform, G(s)alpha-small (45 kDa), was reduced while the biologically inactive isoform, G(s)alpha-large (52 kDa), increased. The increased electrically induced [Ca(2+)](i) transient and cAMP with 10-100 microM forskolin were significantly attenuated in chronically hypoxic rats. The content of G(i)alpha(2), the predominant isoform of G(i) protein in the heart, was unchanged. Results indicate that impaired functions of G(s) protein and adenylyl cyclase cause beta adrenoceptor desensitization. The impaired function of the G(s) protein may be due to reduced G(s)alpha-small and/or increased G(s)alpha-large, which does not result from changes in G(i) protein. Responses to all treatments were the same for right and left ventricles, indicating that the impaired cardiac functions are not secondary to cardiac hypertrophy. PMID- 11029294 TI - Molecular determinants for the distinct pH sensitivity of Kir1.1 and Kir4.1 channels. AB - Kir1.1 (ROMK1) is inhibited by hypercapnia and intracellular acidosis with midpoint pH for channel inhibition (pK(a)) of approximately 6.7. Another close relative, Kir4.1 (BIR10), is also pH sensitive with much lower pH sensitivity (pK(a) approximately 6. 0), although it shares a high sequence homology with Kir1.1. To find the molecular determinants for the distinct pH sensitivity, we studied the structure-functional relationship using site-directed mutagenesis. An NH(2)-terminal residue (Lys-53) was found to be responsible for the low pH sensitivity in Kir4.1. Mutation of this lysine to valine (K53V), a residue seen at the same position in Kir1. 1, markedly increased channel sensitivity to CO(2)/pH. Reverse mutation on Kir1.1 (V66K) decreased the CO(2)/pH sensitivities. Interestingly, mutation of these residues to glutamate greatly enhanced the pH sensitivity in both channels. Other contributors to the distinct pH sensitivity were histidine residues in the COOH terminus, whose numbers are fewer in Kir4.1 than Kir1.1. Mutation of two of these histidine residues in Kir1.1 (H342Q/H354N) reduced CO(2)/pH sensitivities, whereas the creation of two histidines (S328H/G340H) in Kir4.1 increased the CO(2)/pH sensitivities. Combined mutations of the lysine and histidine residues in Kir4.1 (K53V/S328H/G340H) gave rise to a channel that had CO(2)/pH sensitivities almost identical to those of the wild type Kir1.1. Thus the residues demonstrated in our current studies are likely the molecular basis for the distinct pH sensitivity between Kir1.1 and Kir4.1. PMID- 11029295 TI - Differential expression of cell-cell adhesion proteins and cyclin D in MEK1 transdifferentiated MDCK cells. AB - Overexpression of a constitutively active mutant of the mitogen-activated protein kinase kinase MEK1 (caMEK1) in epithelial Madin-Darby canine kidney (MDCK)-C7 cells disrupts morphogenesis, induces an invasive phenotype, and is associated with a reduced rate of cell proliferation. The role of cell-cell adhesion molecules and cell cycle proteins in these processes, however, has not been investigated. We now report loss of E-cadherin expression as well as a marked reduction of beta- and alpha-catenin expression in transdifferentiated MDCK-C7 cells stably expressing caMEK1 (C7caMEK1) compared with epithelial mock transfected MDCK-C7 (C7Mock1) cells. At least part of the remaining alpha-catenin was coimmunoprecipitated with beta-catenin, whereas no E-cadherin was detected in beta-catenin immunoprecipitates. In both cell types, the proteasome-specific protease inhibitors N-acetyl-Leu-Leu-norleucinal (ALLN) and lactacystin led to a time-dependent accumulation of beta-catenin, including the appearance of high molecular-weight beta-catenin species. Quiescent as well as serum-stimulated C7caMEK1 cells showed a higher cyclin D expression than epithelial C7Mock1 cells. The MEK inhibitor U-0126 inhibited extracellular signal-regulated kinase phosphorylation and cyclin D expression in C7caMEK1 cells and almost abolished their already reduced cell proliferation rate. We conclude that the transdifferentiated and invasive phenotype of C7caMEK1 cells is associated with a diminished expression of proteins involved in cell-cell adhesion. Although beta catenin expression is reduced, C7caMEK1 cells show a higher expression of U-0126 sensitive cyclin D protein. PMID- 11029296 TI - Impaired [Ca(2+)](i) and pH(i) responses to kappa-opioid receptor stimulation in the heart of chronically hypoxic rats. AB - kappa-Opioid receptor (kappa-OR) stimulation with U50,488H, a selective kappa-OR agonist, or activation of protein kinase C (PKC) with 4-phorbol 12-myristate 13 acetate (PMA), an activator of PKC, decreased the electrically induced intracellular Ca(2+) ([Ca(2+)](i)) transient and increased the intracellular pH (pH(i)) in single ventricular myocytes of rats subjected to 10% oxygen for 4 wk. The effects of U50,488H were abolished by nor-binaltorphimine, a selective kappa OR antagonist, and calphostin C, a specific inhibitor of PKC, while the effects of PMA were abolished by calphostin C and ethylisopropylamiloride (EIPA), a potent Na(+)/H(+) exchange blocker. In both right hypertrophied and left nonhypertrophied ventricles of chronically hypoxic rats, the effects of U50,488H or PMA on [Ca(2+)](i) transient and pH(i) were significantly attenuated and completely abolished, respectively. Results are first evidence that the [Ca(2+)](i) and pH(i) responses to kappa-OR stimulation are attenuated in the chronically hypoxic rat heart, which may be due to reduced responses to PKC activation. Responses to all treatments were the same for right and left ventricles, indicating that the functional impairment is independent of hypertrophy. kappa-OR mRNA expression was the same in right and left ventricles of both normoxic and hypoxic rats, indicating no regional specificity. PMID- 11029297 TI - Calcium regulates estrogen increase in permeability of cultured CaSki epithelium by eNOS-dependent mechanism. AB - Estrogen increases baseline transepithelial permeability across CaSki cultures and augments the increase in permeability in response to hypertonic gradients. In estrogen-treated cells, lowering cytosolic calcium abrogated the hypertonicity induced augmented increase in permeability and decreased baseline permeability to a greater degree than in estrogen-deprived cells. Steady-state levels of cytosolic calcium in estrogen-deprived cells were higher than in estrogen-treated cells. Increases in extracellular calcium increased cytosolic calcium more in estrogen-deprived cells than in estrogen-treated cells. However, in estrogen treated cells, increasing cytosolic calcium was associated with greater increases in permeability in response to hypertonic gradients than in estrogen-deprived cells. Lowering cytosolic calcium blocked the estrogen-induced increase in nitric oxide (NO) release and in the in vitro conversion of L-[(3)H]arginine to L [(3)H]citrulline. Treatment with estrogen upregulated mRNA of the NO synthase isoform endothelial nitric oxide synthase (eNOS). These results indicate that cytosolic calcium mediates the responses to estrogen and suggest that the estrogen increase in permeability and the augmented increase in permeability in response to hypertonicity involve an increase in NO synthesis by upregulation of the calcium-dependent eNOS. PMID- 11029298 TI - Phenylarsine oxide inhibits heat shock protein 70 induction in cultured guinea pig gastric mucosal cells. AB - Phenylarsine oxide (PAO) forms a stable ring complex with vicinal dithiols that can be reversed with 2,3-dimercaptopropanol (DMP) but not by dithiothreitol (DTT) or 2-mercaptoethanol (2-ME). PAO at 2 microM or higher inhibited heat shock protein 70 (HSP70) induction within minutes in cultured guinea pig gastric mucosal cells exposed to heat (43 degrees C) for 30 min. PAO did not affect the nuclear translocation and phosphorylation of heat shock factor 1 (HSF1) induced by heat stress, but it completely blocked the binding activity of HSF1 to the heat shock element (HSE), leading to the block of expression of HSP70 mRNA and accumulation of HSP70 in the cells. These inhibitions were completely reversed with 2 microM DMP but not with 0.1 mM DTT or 1 mM 2-ME, suggesting specific interactions between PAO and vicinal dithiol-containing molecules. Thioredoxin (Trx) reversed the inhibition of the binding activity of HSF1 in whole cell extracts prepared from PAO-treated, heat-stressed cells. Our results suggest that PAO may react with vicinal-containing molecules including Trx and specifically block the interaction between HSF1 and HSE. PMID- 11029299 TI - Regulation of Na(+)-K(+)-ATPase by cAMP-dependent protein kinase anchored on membrane via its anchoring protein. AB - Na(+)-K(+)- ATPase alpha-subunits in basolateral membrane vesicles (BLMVs) purified from rat parotid glands were (32)P-labeled within 5 s by incubation with [gamma-(32)P]ATP at 37 degrees C in the presence of cAMP, but no labeling occurred without cAMP. Phosphorylation of Na(+)-K(+)-ATPase was associated with a decrease in its activity. This alpha-subunit phosphorylation disappeared when BLMVs were briefly incubated with cAMP and subsequent washing before the incubation with [gamma-(32)P]ATP, indicating that catalytic subunit of protein kinase A (PKA) associated to BLMVs via binding with its RII regulatory subunit anchored on the membrane. In the absence of cAMP, a PKA catalytic subunit readily reassociated with the membrane-bound RII subunit. HT-31 peptide inhibited the Na(+)-K(+)-ATPase phosphorylation by membrane-bound endogenous PKA, indicating an involvement of A-kinase anchoring protein (AKAP). AKAP-150 protein in BLMVs was shown by immunoblotting and an RII overlay assay and was coimmunoprecipitated by anti-RII antibody. These results show that Na(+)-K(+)-ATPase of rat parotid gland acinar cells is regulated in vivo by membrane-anchored PKA via AKAP rather than by free cytosolic PKA. PMID- 11029300 TI - Differential regulation of Ca(2+) sparks and Ca(2+) waves by UTP in rat cerebral artery smooth muscle cells. AB - Uridine 5'-triphosphate (UTP), a potent vasoconstrictor that activates phospholipase C, shifted Ca(2+) signaling from sparks to waves in the smooth muscle cells of rat cerebral arteries. UTP decreased the frequency of Ca(2+) sparks and transient Ca(2+)-activated K(+) (K(Ca)) currents and increased the frequency of Ca(2+) waves. The UTP-induced reduction in Ca(2+) spark frequency did not reflect a decrease in global cytoplasmic Ca(2+), Ca(2+) influx through voltage-dependent Ca(2+) channels (VDCC), or Ca(2+) load of the sarcoplasmic reticulum (SR), since global Ca(2+) was elevated, blocking VDCC did not prevent the effect, and SR Ca(2+) load did not decrease. However, blocking protein kinase C (PKC) with bisindolylmaleimide I did prevent UTP reduction of Ca(2+) sparks and transient K(Ca) currents. UTP decreased the effectiveness of caffeine, which increases the Ca(2+) sensitivity of ryanodine-sensitive Ca(2+) release (RyR) channels, to activate transient K(Ca) currents. This work supports the concept that vasoconstrictors shift Ca(2+) signaling modalities from Ca(2+) sparks to Ca(2+) waves through the concerted actions of PKC on the Ca(2+) sensitivity of RyR channels, which cause Ca(2+) sparks, and of inositol trisphosphate (IP(3)) on IP(3) receptors to generate Ca(2+) waves. PMID- 11029301 TI - Sustained membrane depolarization and pulmonary artery smooth muscle cell proliferation. AB - Pulmonary vasoconstriction and vascular medial hypertrophy greatly contribute to the elevated pulmonary vascular resistance in patients with pulmonary hypertension. A rise in cytosolic free Ca(2+) ([Ca(2+)](cyt)) in pulmonary artery smooth muscle cells (PASMC) triggers vasoconstriction and stimulates cell growth. Membrane potential (E(m)) regulates [Ca(2+)](cyt) by governing Ca(2+) influx through voltage-dependent Ca(2+) channels. Thus intracellular Ca(2+) may serve as a shared signal transduction element that leads to pulmonary vasoconstriction and vascular remodeling. In PASMC, activity of voltage-gated K(+) (Kv) channels regulates resting E(m). In this study, we investigated whether changes of Kv currents [I(K(V))], E(m), and [Ca(2+)](cyt) affect cell growth by comparing these parameters in proliferating and growth-arrested PASMC. Serum deprivation induced growth arrest of PASMC, whereas chelation of extracellular Ca(2+) abolished PASMC growth. Resting [Ca(2+)](cyt) was significantly higher, and resting E(m) was more depolarized, in proliferating PASMC than in growth-arrested cells. Consistently, whole cell I(K(V)) was significantly attenuated in PASMC during proliferation. Furthermore, E(m) depolarization significantly increased resting [Ca(2+)](cyt) and augmented agonist-mediated rises in [Ca(2+)](cyt) in the absence of extracellular Ca(2+). These results demonstrate that reduced I(K(V)), depolarized E(m), and elevated [Ca(2+)](cyt) may play a critical role in stimulating PASMC proliferation. Pulmonary vascular medial hypertrophy in patients with pulmonary hypertension may be partly caused by a membrane depolarization-mediated increase in [Ca(2+)](cyt) in PASMC. PMID- 11029302 TI - Distinct mechanisms of action of selective estrogen receptor modulators in breast and osteoblastic cells. AB - Raloxifene and idoxifene are selective estrogen receptor modulators (SERMs) that exhibit tissue-specific agonist or antagonist properties via interactions with the estrogen receptor (ER). Both compounds are similarly osteoprotective in the ovariectomized rat in vivo as assessed by measurement of bone mineral density, urinary pyridinium cross-links, and serum osteocalcin, suggesting a similar mechanism of action. However, we have identified a fundamental difference in this mechanism via the estrogen response element (ERE) in osteoblast-like cells. With the use of ERE-luciferase reporter constructs, raloxifene, like the complete ER antagonist ICI-182780, acts as an antagonist via the ERE in osteoblastic cells. In contrast, idoxifene, like 17beta-estrogen itself and 4-OH-tamoxifen, acts as an agonist in osteoblastic cells via an ER/ERE-mediated mechanism. Both ICI 182780 and raloxifene inhibited the ERE-dependent agonist activity of 17beta estradiol and idoxifene in osteoblastic cells. In contrast, in breast cells, raloxifene, idoxifene, 4-OH-tamoxifen, and ICI-182780 had no agonist activity and, indeed, raloxifene and idoxifene were potent antagonists of ERE-mediated 17beta-estradiol action, indicating an ERE-dependent mode of action in these cells. Although these SERMs exhibit a similar antagonist activity profile in breast cells, they can be distinguished mechanistically in osteoblastic cells. PMID- 11029303 TI - Effects of unweighting and clenbuterol on myosin light and heavy chains in fast and slow muscles of rat. AB - To investigate the plasticity of slow and fast muscles undergoing slow-to-fast transition, rat soleus (SOL), gastrocnemius (GAS), and extensor digitorum longus (EDL) muscles were exposed for 14 days to 1) unweighting by hindlimb suspension (HU), or 2) treatment with the beta(2)-adrenergic agonist clenbuterol (CB), or 3) a combination of both (HU-CB). In general, HU elicited atrophy, CB induced hypertrophy, and HU-CB partially counteracted the HU-induced atrophy. Analyses of myosin heavy (MHC) and light chain (MLC) isoforms revealed HU- and CB-induced slow-to-fast transitions in SOL (increases of MHCIIa with small amounts of MHCIId and MHCIIb) and the upregulation of the slow MHCIa isoform. The HU- and CB induced changes in GAS consisted of increases in MHCIId and MHCIIb ("fast-to faster transitions"). Changes in the MLC composition of SOL and GAS consisted of slow-to-fast transitions and mainly encompassed an exchange of MLC1s with MLC1f. In addition, MLC3f was elevated whenever MHCIId and MHCIIb isoforms were increased. Because the EDL is predominantly composed of type IID and IIB fibers, HU, CB, and HU-CB had no significant effect on the MHC and MLC patterns. PMID- 11029304 TI - MHC isoform composition and Ca(2+)- or Sr(2+)-activation properties of rat skeletal muscle fibers. AB - Chemically skinned single fibers from adult rat skeletal muscles were used to test the hypothesis that, in mammalian muscle fibers, myosin heavy chain (MHC) isoform expression and Ca(2+)- or Sr(2+)-activation characteristics are only partly correlated. The fibers were first activated in Ca(2+)- or Sr(2+)-buffered solutions under near-physiological conditions, and then their MHC isoform composition was determined electrophoretically. Fibers expressing only the MHC I isoform could be appropriately identified on the basis of either the Ca(2+)- or Sr(2+)-activation characteristics or the MHC isoform composition. Fibers expressing one or a combination of fast MHC isoforms displayed no significant differences in their Ca(2+)- or Sr(2+)-activation properties; therefore, their MHC isoform composition could not be predicted from their Ca(2+)- or Sr(2+) activation characteristics. A large proportion of fibers expressing both fast- and slow-twitch MHC isoforms displayed Ca(2+)- or Sr(2+)-activation properties that were not consistent with their MHC isoform composition; thus both fiber typing methods were needed to fully characterize such fibers. These data show that, in rat skeletal muscles, the extent of correlation between MHC isoform expression and Ca(2+)- or Sr(2+)-activation characteristics is fiber-type dependent. PMID- 11029305 TI - Expression of nucleotide-regulated Cl(-) currents in CF and normal mouse tracheal epithelial cell lines. AB - The dominant route for Cl(-) secretion in mouse tracheal epithelium is via Cl(-) channels different from the cystic fibrosis (CF) transmembrane conductance regulator (CFTR), the channel that is defective in CF. It has been proposed that the use of purinergic agonists to activate these alternative channels in human airways may be beneficial in CF. In the present study, two conditionally immortal epithelial cell lines were established from the tracheae of mice possessing the tsA58 T antigen gene, one of which [MTE18-(-/-)] was homozygous for a knockout of CFTR and the other [MTE7b-(+/-)] heterozygous for CFTR expression. In Ussing chamber studies, amiloride (10(-4) M) and a cocktail of cAMP-activating agents (forskolin, IBMX, and dibutyryl cAMP) resulted in small changes in the short circuit current (I(sc)) and resistance of both cell lines, with larger increases in I(sc) being elicited by ionomycin (10(-6) M). Both cell lines expressed P(2)Y(2) receptors and responded to the purinergic agonists ATP, UTP, and 5' adenylylimidodiphosphate (10(-4) M) with an increase in I(sc). This response could be inhibited by DIDS and was abolished in the presence of Cl(-)-free Ringer solution. Reducing the mucosal Cl(-) concentration increased the response to UTP of both cell lines, with a significantly greater increase in MTE18-(-/-) cells. Pretreatment of these cells with thapsigargin caused a direct increase in I(sc) and inhibited the response to UTP. These data suggest that both cell lines express purinergic-regulated Cl(-) currents and may prove valuable tools in studying the properties of this pathway. PMID- 11029306 TI - Amino acid deprivation induces translation of branched-chain alpha-ketoacid dehydrogenase kinase. AB - Leucine, isoleucine, and valine are used by cells for protein synthesis or are catabolized into sources for glucose and lipid production. These branched-chain amino acids influence proteolysis, hormone release, and cell cycle progression along with their other metabolic roles. The branched-chain amino acids play a central role in regulating cellular protein turnover by reducing autophagy. These essential amino acids are committed to their catabolic fate by the activity of the branched-chain alpha-ketoacid dehydrogenase complex. Activity of the branched chain alpha-ketoacid dehydrogenase complex is regulated by phosphorylation/inactivation of the alpha-subunit performed by a complex specific kinase. Here we show that elimination of the branched-chain amino acids from the medium of cultured cells results in a two- to threefold increased production of the branched-chain alpha-ketoacid dehydrogenase kinase with a decrease in the activity state of the branched-chain alpha-ketoacid dehydrogenase complex. The mechanism cells use to increase kinase production under these conditions involves recruitment of the kinase mRNA into polyribosomes. Promoter activity and the steady-state concentration of the mRNA are unchanged by these conditions. PMID- 11029307 TI - Ca(2+)-ATPase protein expression in mammary tissue. AB - Protein expression of plasma membrane Ca(2+)-ATPases (PMCAs) and the putative Golgi secretory pathway Ca(2+)-ATPase (SPCA) was examined in rat mammary tissue. As lactation started, PMCA protein expression increased dramatically, and this increased expression paralleled milk production. Mammary PMCA was primarily PMCA2b but was approximately 4,000 daltons larger than expected. RT-PCR showed that the primary mammary PMCA2b transcript was alternatively spliced, at splice site A, to include an additional 135 bp, resulting in the insertion of 45 amino acids. This splice form is designated 2bw. PMCA2bw is secreted into milk, associated with the milk fat globule membrane. Therefore, PMCA2bw is located on the apical membrane of the secretory cell. Smaller amounts of PMCA1b and 4b protein were found in mammary tissue. PMCA4b was the major PMCA expressed in developing tissue, and its level declined as lactation started. PMCA1b expression increased moderately during lactation. SPCA protein expression increased 1 wk before parturition and increased further as lactation proceeded. The abundance and cell location of PMCA2b suggest that it is important for macro-Ca(2+) homeostasis in lactating tissue. The pattern of expression and abundance of SPCA suggest that it is a candidate for the Golgi Ca(2+)-ATPase. PMID- 11029308 TI - Endogenous nitric oxide is implicated in the regulation of lipolysis through antioxidant-related effect. AB - We studied the influence of nitric oxide (NO) endogenously produced by adipocytes in lipolysis regulation. Diphenyliodonium (DPI), a nitric oxide synthase (NOS) inhibitor, was found to completely suppress NO synthesis in intact adipocytes and was thus used in lipolysis experiments. DPI was found to decrease both basal and dibutyryl cAMP (DBcAMP)-stimulated lipolysis. Inhibition of DBcAMP-stimulated lipolysis by DPI was prevented by S-nitroso-N-acetyl-penicillamine (SNAP), a NO donor. This antilipolytic effect of DPI was also prevented by two antioxidants, ascorbate or diethyldithiocarbamic acid (DDC). Preincubation of isolated adipocytes with DPI (30 min) before exposure to DBcAMP almost completely abolished the stimulated lipolysis. Addition of SNAP or antioxidant during DPI preincubation restored the lipolytic response to DBcAMP, whereas no preventive effects were observed when these compounds were added simultaneously to DBcAMP. Exposure of isolated adipocytes to an extracellular generating system of oxygen species (xanthine/xanthine oxidase) or to H(2)O(2) also resulted in an inhibition of the lipolytic response to DBcAMP. H(2)O(2) or DPI decreased cAMP-dependent protein kinase (PKA) activation. The DPI effect on PKA activity was prevented by SNAP, ascorbate, or DDC. These results provide clear evidence that 1) the DPI antilipolytic effect is related to adipocyte NOS inhibition leading to PKA alterations, and 2) endogenous NO is required for the cAMP lipolytic process through antioxidant-related effect. PMID- 11029309 TI - Role of MARCKS in regulating endothelial cell proliferation. AB - Myristoylated alanine-rich C kinase substrate (MARCKS), as a specific protein kinase C (PKC) substrate, mediates PKC signaling through its phosphorylation and subsequent modification of its association with filamentous actin (F-actin) and calmodulin (CaM). PKC has long been implicated in cell proliferation, and recent studies have suggested that MARCKS may function as a cell growth suppressor. Therefore, in the present study, we investigated MARCKS protein expression, distribution, and phosphorylation in preconfluent and confluent bovine pulmonary microvascular endothelial cells (BPMEC) in the presence or absence of the vascular endothelial growth factor (VEGF). In addition, we examined functional alterations of MARCKS in these cells by studying the association of MARCKS with F actin and CaM-dependent myosin light chain (MLC) phosphorylation. Our results indicate that MARCKS protein is downregulated during BPMEC proliferation. Decreased MARCKS association with F-actin, increased actin polymerization, and CaM-dependent MLC phosphorylation appear to mediate cell shape changes and motility during BPMEC growth. In contrast, VEGF stimulated MARCKS phosphorylation without alteration of protein expression during BPMEC proliferation, which may result in reduced interaction between MARCKS and actin or CaM, leading to actin reorganization and MLC phosphorylation. Our data suggest a regulatory role of MARCKS during endothelial cell proliferation. PMID- 11029310 TI - Cation transport and cell volume changes in maturing rat reticulocytes. AB - During maturation, reticulocytes lose membrane material, including transporters, and this is accompanied by a loss of cell water and volume. Here we determined a possible role of ion transport in adjusting cell volume during maturation. Reticulocytes and red blood cells of different ages were prepared from erythropoietin-treated rats by density gradient fractionation. Cell volume and ion transport were measured in freshly prepared cells and in reticulocytes during in vitro maturation. Reticulocytes had an increased K content and cell volume, whereas intracellular Na was decreased. All parameters approached whole blood values after 2 days in culture. Na-K pump was elevated in reticulocytes and decreased during maturation. Na-K-2Cl cotransport (NKCC) activity was lower in reticulocytes and was activated 8- and 20-fold by shrinkage and okadaic acid, respectively, whereas stimulation was barely detectable in high-buoyant density red blood cells. The ouabain- and bumetanide-insensitive Na flux in reticulocytes decreased on maturation. Most of it was inhibited by amiloride, indicating the presence of Na/proton exchange. Our results show that, although the Na-K-pump activity in reticulocytes is very much increased, the enhanced capacity of NKCC is essentially cryptic until stimulated. Both types of capacities (activities) decrease during maturation, indicating a possible loss of transport protein. The decrease was constrained to the period of reticulocyte maturation. Loss of transport capacity appears to exceed the loss of membrane surface area. Reticulocyte age-related changes in the net electrochemical driving force indicate that the increasing NKCC activity might contribute to the reduction in cell water. PMID- 11029311 TI - In squid nerves intracellular Mg(2+) promotes deactivation of the ATP-upregulated Na(+)/Ca(2+) exchanger. AB - We investigated the role of intracellular Mg(2+) (Mg(i)(2+)) on the ATP regulation of Na(+)/Ca(2+) exchanger in squid axons and bovine heart. In squid axons and nerve vesicles, the ATP-upregulated exchanger remains activated after removal of cytoplasmic Mg(2+), even in the absence of ATP. Rapid and complete deactivation of the ATP-stimulated exchange occurs upon readmission of Mg(i)(2+). At constant ATP concentration, the effect of intracellular Mg(2+) concentration ([Mg(2+)](i)) on the ATP regulation of exchanger is biphasic: activation at low [Mg(2+)](i), followed by deactivation as [Mg(2+)](i) is increased. No correlation was found between the above results and the levels of phosphatidylinositol 4 phosphate and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] measured in nerve membrane vesicles. Incorporation of PtdIns(4,5)P(2) into membrane vesicles activates Na(+)/Ca(2+) exchange in mammalian heart but not in squid nerve. Moreover, an exogenous phosphatase prevents MgATP activation in squid nerves but not in mammalian heart. It is concluded that 1) Mg(i)(2+) is an essential cofactor for the deactivation part of ATP regulation of the exchanger and 2) the metabolic pathway of ATP upregulation of the Na(+)/Ca(2+) exchanger is different in mammalian heart and squid nerves. PMID- 11029312 TI - Elevated ceramide is downstream of altered calcium homeostasis in low serum induced apoptosis. AB - Two immortalized cell lines, sup (+) and sup (-), derived from mutagenized Syrian hamster embryo cells, were used to study the relationship and temporal order between calcium and ceramide signals during apoptosis. The early preneoplastic cells, termed sup (+), suppress tumorigenicity when hybridized with tumor cells, whereas later-stage sup (-) cells do not. In reduced serum conditions, sup (+) cells cease proliferating and undergo apoptosis; in contrast, sup (-) cells continue slow growth and undergo necrosis. In sup (+) cells, decreased endoplasmic reticulum (ER) calcium occurs 4 h after low serum treatment and precedes apoptosis. Significant elevations in ceramide are observed 16 h after reduced serum treatment of sup (+) cells but are not found in sup (-) cells. Inhibiting ER calcium depletion in low serum-treated sup (+) cells by treating with high levels of calcium prevents both ceramide generation and apoptosis. Conversely, inducing ER calcium depletion in sup (-) cells by treating with low serum plus thapsigargin results in elevated ceramide levels and apoptosis. Furthermore, C(6)-ceramide treatment induced apoptosis of sup (-) cells in low serum, a condition that does not normally cause apoptosis. C(6)-ceramide treatment did not induce apoptosis in either sup (+) or sup (-) cells in 10% serum but did cause G(2)/M arrest. These studies show that ceramide production is downstream of ER calcium release. PMID- 11029313 TI - Expression and localization of Na(+)-HCO(3)(-) cotransporter in bovine corneal endothelium. AB - Functional studies support the presence of the Na(+)-HCO(3)(-) cotransporter (NBC) in corneal endothelium and possibly corneal epithelium; however, molecular identification and membrane localization have not been reported. To test whether NBC is expressed in bovine cornea, Western blotting was performed, which showed a single band at approximately 130 kDa for freshly isolated and cultured endothelial cells, but no band for epithelium. Two isoforms of NBC have recently been cloned in kidney (kNBC) and pancreas (pNBC). RT-PCR was run using cultured and fresh bovine corneal endothelial and fresh corneal epithelial total RNA and specific primers for kNBC and pNBC. RT-PCR analysis for pNBC was positive in endothelium and weak in epithelium. The RT-PCR product was subcloned and confirmed as pNBC by sequencing. No specific bands for kNBC were obtained from corneal cells. Indirect immunofluorescence and confocal microscopy indicated that NBC locates predominantly to the basolateral membrane in corneal endothelial cells. Furthermore, Na(+)-dependent HCO(3)(-) fluxes and HCO(3)(-)-dependent cotransport with Na(+) were elicited only from the basolateral side of corneal endothelial cells. Therefore, we conclude that pNBC is present in the basolateral membrane of both fresh and cultured bovine corneal endothelium and weakly expressed in the corneal epithelium. PMID- 11029314 TI - Smooth muscle myosin light chain kinase expression in cardiac and skeletal muscle. AB - The purpose of this study was to characterize myosin light chain kinase (MLCK) expression in cardiac and skeletal muscle. The only classic MLCK detected in cardiac tissue, purified cardiac myocytes, and in a cardiac myocyte cell line (AT1) was identical to the 130-kDa smooth muscle MLCK (smMLCK). A complex pattern of MLCK expression was observed during differentiation of skeletal muscle in which the 220-kDa-long or "nonmuscle" form of MLCK is expressed in undifferentiated myoblasts. Subsequently, during myoblast differentiation, expression of the 220-kDa MLCK declines and expression of this form is replaced by the 130-kDa smMLCK and a skeletal muscle-specific isoform, skMLCK in adult skeletal muscle. These results demonstrate that the skMLCK is the only tissue specific MLCK, being expressed in adult skeletal muscle but not in cardiac, smooth, or nonmuscle tissues. In contrast, the 130-kDa smMLCK is ubiquitous in all adult tissues, including skeletal and cardiac muscle, demonstrating that, although the 130-kDa smMLCK is expressed at highest levels in smooth muscle tissues, it is not a smooth muscle-specific protein. PMID- 11029316 TI - Authors, authors, authors-follow instructions or expect delay PMID- 11029317 TI - In vivo lipid peroxidation and platelet activation in cystic fibrosis. AB - F(2)-isoprostanes are bioactive peroxidation products of arachidonic acid whose urinary excretion provides an index of lipid peroxidation in vivo. We tested the hypothesis that formation of F(2)-isoprostanes is altered in patients with cystic fibrosis and contributes to platelet activation and pulmonary dysfunction in this setting. The urinary excretion of immunoreactive 8-iso-prostaglandin F(2alpha) (PGF(2alpha)) was significantly (p = 0.0001) higher in 36 patients with cystic fibrosis than in 36 age-matched healthy subjects: 618 +/- 406 versus 168 +/- 48 pg/mg creatinine. The urinary excretion of immunoreactive 11-dehydro-thromboxane B(2) (TXB(2)), an index of in vivo platelet activation, was also significantly (p = 0.0001) higher in patients than in control subjects: 2,440 +/- 1,453 versus 325 +/- 184 pg/mg creatinine. The excretion rate of 8-iso-PGF(2alpha) was correlated with that of 11-dehydro-TXB(2) (rho = 0.51; p = 0.0026) and inversely related to FEV(1) (rho = -0.40; p = 0.0195). Urinary 8-iso-PGF(2alpha) excretion was largely unaffected during cyclooxygenase inhibition with low-dose aspirin, nimesulide, or ibuprofen, consistent with a noncyclooxygenase mechanism of F(2)-isoprostane formation in cystic fibrosis. Increased vitamin E supplementation (from 200 to 600 mg/d) was associated with statistically significant (p = 0.005) reductions in urinary 8-iso-PGF(2alpha) and 11-dehydro-TXB(2) excretion, by 42% and 29%, respectively. We conclude that enhanced lipid peroxidation is an important feature of cystic fibrosis and may contribute to persistent platelet activation and pulmonary dysfunction via generation of bioactive isoeicosanoids. Our results provide a rationale for reassessing the adequacy of vitamin E supplementation in this setting. PMID- 11029315 TI - beta-adrenergic receptor/cAMP-mediated signaling and apoptosis of S49 lymphoma cells. AB - beta-Adrenergic receptor (betaAR) activation and/or increases in cAMP regulate growth and proliferation of a variety of cells and, in some cells, promote cell death. In the current studies we addressed the mechanism of this growth reduction by examining betaAR-mediated effects in the murine T-lymphoma cell line S49. Wild type S49 cells, derived from immature thymocytes (CD4(+)/CD8(+)) undergo growth arrest and subsequent death when treated with agents that increase cAMP levels (e.g., betaAR agonists, 8-bromo-cAMP, cholera toxin, forskolin). Morphological and biochemical criteria indicate that this cell death is a result of apoptosis. In cyc(-) and kin(-) S49 cells, which lack G(s)alpha and functional protein kinase A (PKA), respectively, betaAR activation of G(s)alpha and cAMP action via PKA are critical steps in this apoptotic pathway. S49 cells that overexpress Bcl 2 are resistant to cAMP-induced apoptosis. We conclude that betaAR activation induces apoptosis in immature T lymphocytes via G(s)alpha and PKA, while overexpression of Bcl-2 prevents cell death. betaAR/cAMP/PKA-mediated apoptosis may provide a means to control proliferation of immature T cells in vivo. PMID- 11029318 TI - Effect of isocapnic hypoxia on variational activity of breathing. AB - In the presence of either hypocapnia or sleep, hypoxia has been shown to induce periodic breathing and increase the total variational activity of breath components. It is not known whether hypoxia induces alterations in breathing variability during wakefulness and in the absence of hypocapnia. To address this issue, we studied nonobtrusively 14 healthy awake subjects before and during the delivery of a hypoxic gas mixture via a plastic hood; the subjects' oxygen saturation decreased from 98 to 79% and end-tidal carbon dioxide tension was kept constant. Compared with air, isocapnic hypoxia increased the gross variability of minute ventilation (V I), tidal volume (VT), inspiratory time (TI), and expiratory time (TE) (all p < 0.004). Isocapnic hypoxia decreased the autocorrelation coefficient at a lag of one breath for TE (p < 0. 008) and V I (p = 0.07), the number of consecutive breath lags having significant autocorrelation coefficients for TE (p = 0.03), and the cycle time of oscillations in V I (p = 0.03). When partitioned, the increase in total variational activity during isocapnic hypoxia was found to result from increases in the random fractions of V I, VT, TI, and TE (all p < 0.05), and the oscillatory fractions of V I, VT, and TE (all p < 0.03). In conclusion, hypoxia induced hidden oscillations in V I, VT, and TE despite wakefulness and an isocapnic state, suggesting that neural responses may have a more important role in the genesis of hypoxia-induced oscillations than previously reported. PMID- 11029319 TI - Frequency and significance of increased upper airway resistance during sleep. AB - Previous studies have shown that episodes of inspiratory flow limitation can be clinically important. We have challenged the hypothesis that patients with the "upper airway resistance syndrome" have more episodes of inspiratory flow limitation, associated with greater swings in pleural pressure and more arousals from sleep than normal subjects. We thus studied eight symptomatic patients with UARS (ESS > 10, AHI < 15) and eight matched asymptomatic subjects. All had overnight polysomnography with recording of pleural pressure and airflow derived from nasal pressure. Events scored visually using nasal pressure were termed flow limitation events and those using both signals "resistive events." The patients with UARS had no more episodes of flow limitation or resistive events than the controls. However, pleural pressure swings at resistive event termination were significantly more negative in the patient group (-15 [IQR 9-19]; -11 [8-12] cm H(2)O; p = 0.02) and the number of cortical arousals associated with resistive events was higher in the patients (median, 10 [5-15]; 3 [1-9]/h slept; p = 0.02). This study confirms that patients with UARS have periods of increased upper airway resistance associated with significant sleep fragmentation. However, resistive events are also common in normal subjects, although these are associated with less negative pleural pressure swings and fewer arousals. Thus, the clinical significance of resistive events needs to be interpreted with caution. PMID- 11029320 TI - Use of the child health questionnaire in a sample of moderate and low-income inner-city children with asthma. AB - The Child Health Questionnaire (CHQ-PF50) is one of several recent efforts to gauge pediatric, health-related quality of life from the patient's (or parent's) perspective. Although tested extensively with healthy children, more information is needed about CHQ performance among children with chronic conditions such as asthma. The current study extends previous work by examining the CHQ's psychometric performance in a sample of children with asthma, overrepresenting those at high risk for poor outcomes. Seventy-four adult caregivers of children with asthma completed the CHQ. Internal consistency reliability was consistently high for all but one scale. Intraclass correlation coefficients ranged from a low of 0.37 to a high of 0.84. Tests of validity found CHQ scales better at distinguishing levels of disease severity as defined by symptom activity than medication use or pulmonary function tests. Performance of the CHQ-PF50 in a sample of low-income to moderate income inner-city parents of children with asthma presented mixed results. The instrument addresses a broad range of concepts but some scales may be more salient than others in assessing health status of children at highest risk for asthma morbidity. Future efforts must compare condition-specific and generic instruments to evaluate their relative strengths and weakness, as well as potential links between them. PMID- 11029321 TI - Incidence and determinants of IgE-mediated sensitization in apprentices. A prospective study. AB - We investigated prospectively the incidence and determinants of work-related specific skin sensitization in a cohort of 769 apprentices, including 417 in animal health technology, 230 in pastry-making, and 122 in dental-hygiene technology. Subjects were recruited when starting exposure to laboratory animals, flour, or latex. A questionnaire and skin-prick tests with common and work related allergens were administered on entry and at follow-up visits from 8 to 44 mo; information on number of hours of exposure to specific allergens was obtained. Among 769 apprentices, 698 attended >/= 1 follow-up visit. A total of 111 subjects developed specific sensitization over the study period. The incidence of work-related sensitization (per person-year) was 8.9% (95% CI 7.3 to 11.0%) in the animal-health program, 4.2% (95% CI 1.8 to 8.2%) in the pastry making program, and 2.5% (95% CI = 0.7 to 4.3%) in the dental-hygiene program. In the animal health group, Cox regression analyses showed that atopy, nasal, and respiratory symptoms in the pollen season, and exposure assessed by the school attended or by duration of exposure to rodents were the most significant predictors of sensitization. In the dental-hygiene program, atopy and asthma were significant determinants. This study shows that: (1) an apprenticeship in animal health technology carries a greater risk of developing specific sensitization than do apprenticeships in pastry-making and dental-hygiene; (2) atopy, respiratory symptoms in the pollen season, and number of hours in contact with rodents determine the risk of sensitization in apprentices in the animal health program. PMID- 11029323 TI - Deletion polymorphisms in the angiotensin converting enzyme gene are associated with pulmonary hypertension evoked by exercise challenge in patients with chronic obstructive pulmonary disease. AB - Angiotensin converting enzyme (ACE) plays an important role in the pathogenesis of pulmonary hypertension. In this study we determined whether the deletion (D)/insertion (I) polymorphism in the ACE gene may be associated with pulmonary hypertension evoked by exercise challenge in patients with chronic obstructive pulmonary disease (COPD). ACE genotypes were determined in 19 patients with COPD. All patients underwent right heart catheterization followed by a constant-load exercise test while breathing room air or oxygen. Subgroups were created of seven patients with the II genotype, six with the ID genotype, and six with the DD genotype who were well-matched with respect to age, blood gas data at rest or after exercise, baseline lung function, results of incremental exercise testing, and hemodynamic data at rest. The mean pulmonary arterial pressure (Ppa) and pulmonary vascular resistance (Rpv) at rest in the three subgrpoups did not differ significantly during breathing of either room air or oxygen. However, the Ppa after exercise challenge in patients with the DD genotype (55.7 +/- 4.9 mm Hg [mean +/- SD]) was significantly higher than in patients with the II genotype (42.6 +/- 7.1 mm Hg, p = 0.008). The Rpv after exercise in patients with the DD genotype was also significantly higher than in patients with the ID and II genotypes. During breathing of oxygen to diminish acute hypoxic pulmonary vasoconstriction, the Ppa in patients with the DD genotype (52.3 +/- 3.1 mm Hg) was higher than in patients with the ID genotype (40.5 +/- 5.9 mm Hg, p = 0.0049) or the II genotype (37.7 +/- 5.9 mm Hg, p = 0.0027). In addition, the Rpv in patients with the DD genotype was higher than in patients with the ID and II genotypes. These results suggest that D-I polymorphism in the ACE gene may be associated with pulmonary hypertension evoked by exercise challenge in patients with COPD. However, the number of patients in this study was very small for a genetic association study, and our results should be examined in larger studies. PMID- 11029322 TI - Effects of budesonide and formoterol on eosinophil activation induced by human lung fibroblasts. AB - Budesonide and formoterol are extensively used in current asthma therapy. Budesonide is known as potent antiinflammatory agent and formoterol also appears to have some antiinflammatory properties. We investigated inhibitory effects of these drugs on eosinophil activation in vitro as induced by fibroblast conditioned medium (FCM). We measured the modulation of expression of clonal designator (CD)11b and L-selectin with flow cytometry after 4 h or 16 h of culture of eosinophils when budesonide or formoterol was applied either directly to the eosinophils while they were stimulated with FCM (direct method) or when each drug was applied to lung fibroblasts from which conditioned medium was then administered to eosinophils (indirect method). In the direct method, budesonide (10(-)(8) M) inhibited the modulation of CD11b (44 [25th to 75th percentiles: 26 to 66]% of control) and L-selectin (30 [-13 to 48]% of control) only after 16 h, and not after 4 h. Formoterol did not directly inhibit the modulation of eosinophil CD11b and L-selectin expression. In the indirect method, both budesonide and formoterol inhibited lung fibroblast activation, resulting in diminished eosinophil activation after 4 h. Budesonide or formoterol at 10(-)(8) M inhibited upregulation of CD11b to 26 [15 to 40]% and 38 [23 to 46]%, respectively, and inhibited L-selectin shedding to 14 [-3 to 50]% and 27 [2 to 62]%, respectively, of control values. These results show that budesonide inhibits eosinophil activation primarily through effects on lung fibroblasts, presumably by inhibiting production of granulocyte-macrophage colony-stimulating factor. After longer incubation periods, budesonide also directly inhibits eosinophil activation. In contrast, formoterol can inhibit eosinophil activation only via inhibitory effects on lung fibroblasts. We did not observe an additional effect of formoterol, beyond the effects induced by budesonide under any circumstance studied. Lung fibroblasts, in addition to eosinophils, may serve as important target cells for antiinflammatory treatment in asthma. PMID- 11029324 TI - Disturbances in leptin metabolism are related to energy imbalance during acute exacerbations of chronic obstructive pulmonary disease. AB - Previously we reported an impaired energy balance in patients with chronic obstructive pulmonary disease (COPD) during an acute disease exacerbation, but limited data are available on the underlying mechanisms. Experimental and clinical research supports the hypothesis of involvement of the hormone leptin in body weight and energy balance homeostasis. The aim of this study was to investigate the course of the energy balance in relation to leptin and the soluble tumor necrosis factor (TNF) receptors (sTNF-R) 55 and 75, plasma glucose, and serum insulin in patients with severe COPD during the first 7 d of hospitalization for an acute exacerbation (n = 17, 11 men, age mean [SD] 66 [10] yr, FEV(1) 36 [12] %pred). For reference values of the laboratory parameters, blood was collected from 23 (16 men) healthy, elderly subjects. On admission, the dietary intake/resting energy expenditure (REE) ratio was severely depressed (1.28 [0.57]), but gradually restored until Day 7 (1.65 [0. 45], p = 0.005 versus Day 1). Glucose and insulin concentrations were elevated on admission, but on Day 7 only plasma glucose was decreased. The sTNF-Rs were not different from healthy subjects and did not change. Plasma leptin, adjusted for fat mass expressed as percentage of body weight (%FM), was elevated on Day 1 compared with healthy subjects (1.82 [3.85] versus 0.32 [0.72] ng%/ml, p = 0.008), but decreased significantly until Day 7 (1.46 [3.77] ng%/ml, p = 0. 015 versus Day 1). On Day 7, sTNF-R55 was, independently of %FM, correlated with the natural logarithm (LN) of leptin (r = 0.65, p = 0.041) and with plasma glucose (r = 0.81, p = 0.015). In addition, the dietary intake/REE ratio was not only inversely related with LN leptin (-0.74, p = 0.037), but also with sTNF-R55 (r = -0.93, p = 0. 001) on day seven. In conclusion, temporary disturbances in the energy balance were seen during an acute exacerbation of COPD, related to increased leptin concentrations as well as to the systemic inflammatory response. Evidence was found that the elevated leptin concentrations were in turn under control of the systemic inflammatory response, and, presumably, the high-dose systemic glucocorticosteroid treatment. PMID- 11029325 TI - Respiratory symptoms of obstructive lung disease in European crop farmers. AB - Crop farming as a risk factor for respiratory symptoms of obstructive lung disease was assessed. Random samples of crop farmers from four European countries were studied following a cross-sectional design. A questionnaire on respiratory symptoms and occupation was administered to determine prevalences, and the roles of the various crops as risk factors for respiratory symptoms were assessed through logistic regression modeling. The 4,793 crop farmers included in the study (response rate: 85.3%) reported the following respiratory symptoms: wheezing (14.9%), asthma (3.3%), nasal allergy (14.4%), chronic phlegm (12.4%), organic dust toxic syndrome (ODTS) (15.2%), and symptoms at work (22.0%). In the multivariate analysis, adjusting for age, sex, smoking, country, and exposure to other plants or livestock, flower growing was a risk factor for asthma (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.1-3.9) and cultivating oil plants was associated with ODTS (OR 1.5, 95% CI 1.3-1.9), symptoms at work (OR 1.4, 95% CI 1.2- 1.7), and chronic phlegm (OR 1.3, 95% CI 1.1-1.6). Working inside greenhouses was a marginal risk factor for asthma (OR 2.1, 95% CI 0.9-4.5). We conclude that flower and oil plant production is associated with increased risk of respiratory symptoms in European crop farmers. PMID- 11029326 TI - Exogenous surfactant supplementation in infants with respiratory syncytial virus bronchiolitis. AB - Infants with respiratory syncytial virus (RSV) bronchiolitis are deficient in surfactant, both in quantity and ability to reduce surface tension. New evidence suggests surfactant has a role in maintaining the patency of conducting airways, which has implications for RSV bronchiolitis. A randomized, controlled pilot study was undertaken to assess the effects of exogenous surfactant supplementation to RSV-positive infants on pulmonary mechanics, indices of gas exchange, and the phospholipid composition of bronchoalveolar lavage fluid (BALF). Nineteen ventilated infants (median corrected age 4 wk) received either two doses of surfactant (Survanta, 100 mg/kg) within 24 and 48 h of mechanical ventilation (n = 9), or air placebo (n = 10). Static lung compliance and resistance of infants in the placebo but not in the surfactant-treated group became progressively worse over the first 30 h following enrollment. Although no significant acute changes in gas exchange parameters were seen following surfactant, infants in the surfactant group showed a more rapid improvement in oxygenation and ventilation indices over the first 60 h of ventilation. Surfactant status was assessed from the concentration ratio in BALF of the disaturated phospholipid species dipalmitoylphosphatidylcholine to that of the monounsaturated species palmitoyloleoylphosphatidylcholine. This ratio correlated with both lung compliance (positively) and resistance (negatively), and over time increased in the treated group and declined in placebo infants. The data from this pilot study suggest that functional surfactant has a role in maintaining small airway patency as well as lung compliance in infants infected with RSV and an outcome study is now warranted. PMID- 11029327 TI - Interrelation between oxygen tension and nitric oxide in the respiratory system. AB - To understand the relationship between oxygen tension and nitric oxide (NO) function, one animal and two human studies were designed. In the animal study, the effect of NO in inducing the relaxation of aortic specimens was significantly lower by 68% under 480 mm Hg of oxygen tension than under 28 mm Hg, indicating that oxygen tension has an important role in determining the biological effects of NO. In a clinical analysis with nonsmokers (n = 23), the alveolar-to-arterial difference for oxygen (A-aDO(2)) was reciprocally correlated with exhaled NO concentrations (r = 0.53). Because NO concentration in the lower respiratory zone depends partly on the amount of inspirable NO originating in the upper airway, a well-ventilated area, requiring much perfusion, could receive greater amounts of NO than a poorly ventilated one. Thus, the reciprocal relation of A-aDO(2) with the concentration of exhaled NO is not necessarily incompatible with the effect of hypoxic pulmonary vasoconstriction in ventilation-to-perfusion (V'A/Q') imbalance. In our third experiment, with nonsmokers (n = 21), pure oxygen inhalation during mechanical ventilation significantly decreased the concentration of exhaled NO and enhanced A-aDO(2), indicating a relationship between NO and oxygen similar to that observed in the animal experiment. These findings led us to conclude that a positive relation between exhaled NO and blood oxygenation efficiency exists in the respiratory system, and further, that oxygen might affect this relationship. Thus, the relative balance of NO and oxygen concentrations may be another factor for consideration in respiratory function. PMID- 11029328 TI - Exhaled nitric oxide production by nitric oxide synthase-deficient mice. AB - Nitric oxide (NO) is produced in the nasal cavities, airways, and lungs and is exhaled by normal animals and humans. Although increased exhaled NO concentrations in airway inflammation have been associated with increased airway expression of nitric oxide synthase 2 (NOS 2), it is uncertain which NOS isoform is responsible for baseline levels of exhaled NO. We therefore studied wild-type mice and mice with a congenital deficiency of NOS 1, NOS 2, or NOS 3. By studying a closed chamber in which the exhaled gas of a group of mice was collected, gaseous NO production rates were measured. Wild-type mice exhaled 362 +/- 35 x 10(-15) mol g(-1) min(-1) NO (mean +/- SE, n = 16 groups of five mice), NOS 1 deficient mice exhaled 592 +/- 74 x 10(-15) mol g(-1) min(-1) NO (n = 15 groups, p < 0.05 versus wild-type and NOS 2-deficient mice), NOS 2-deficient mice 330 +/- 74 x 10(-15) mol g(-1) min(-1) NO (n = 14 groups) and NOS 3-deficient mice 766 +/ 101 x 10(-15) mol g(-1) min(-1) NO (n = 16 groups, p < 0.001 versus wild-type and NOS 2-deficient mice). Pharmacological NOS inhibition with L-NAME decreased (p < 0.05) the exhaled NO production rate of wild-type and NOS 3-deficient but not of NOS 2-deficient mice. L-Arginine administration increased exhaled NO production rate in all but NOS 2-deficient mice. Absence of NOS 1 or 3 is associated with increased murine exhaled NO production rates. Since NOS 2 deficient mice were the only genotype to lack substrate- and inhibitor-regulated changes of NO exhalation, we suggest that NOS 2 is an important isoform contributing to exhaled NO exhalation in healthy mice. PMID- 11029329 TI - Linkage analysis of markers on chromosome 11q13 with asthma and atopy in a United Kingdom population. AB - Previous studies have suggested that atopy is linked to the beta chain of the high affinity IgE receptor (Fcepsilon R1-beta) on chromosome 11q13. Fcepsilon R1 beta polymorphisms, I181L, V183L, and E237G, are reported to be associated with asthma and atopy. The aim of this study was to investigate linkage to Fcepsilon R1-beta in a UK population and to assess the frequency of the polymorphisms and their association with asthma and atopy. A sample of 131 families was recruited at random with a sample of 109 families ascertained via an asthmatic proband. Each subject completed a written and video-assisted questionnaire and underwent bronchial challenge and skin prick testing. Serum total and specific IgE levels were measured. Quantitative scores were derived for asthma and atopy using principal component analysis. Four microsatellite markers were genotyped, including Fcepsilon R1-beta. The frequency of the I181L and V183L polymorphisms were determined by sequencing, and the E237G polymorphism was determined using the amplification refractory mutation system. We found no evidence for linkage to Fcepsilon R1-beta and only weak evidence for linkage to the less informative marker E237G. We found no examples of the I181L/V183L polymorphism in our population sample. Our study has failed to strengthen the evidence for a candidate gene on chromosome 11q13. PMID- 11029330 TI - Increased nitrotyrosine in exhaled breath condensate of patients with asthma. AB - The reaction of nitric oxide (NO) and superoxide anions (O(2)(-)) in the airway results in the formation of peroxynitrite, a highly reactive oxidant species. Peroxynitrite reacts with tyrosine residues in proteins to form the stable product nitrotyrosine. We investigated whether nitrotyrosine in exhaled breath condensates may be increased in patients with asthma. Four groups of nonsmoking subjects were studied. We measured exhaled NO, nitrotyrosine, and leukotrienes concentrations in breath condensate in healthy nonatopic subjects (n = 15) and in patients with mild asthma (steroid naive, n = 15), moderate asthma (inhaled steroid treatment, n = 12), and severe asthma (oral steroid treatment, n = 12). Exhaled NO was increased significantly in patients with mild (19.2 +/- 2.7 ppb, p < 0.01) and moderate asthma (14.0 +/- 1.53 ppb, p < 0.05), as compared with normal control (6.58 +/- 0.61 ppb). The levels of LTC(4)/D(4)/E(4) and LTB(4) were increased significantly in patients with moderate and severe asthma treated with steroids. Nitrotyrosine concentrations were detectable (6.3 +/- 0.8 ng/ml) in breath condensate of normal subjects, and were increased significantly in patients with mild asthma (15.3 +/- 2.0 ng/ml, p < 0.01). However, the levels of nitrotyrosine in exhaled condensate were lower in patients with moderate (5.0 +/- 0.6 ng/ml) and severe asthma (3.3 +/- 0.6 ng/ml, p < 0.05). There was a significant correlation between nitrotyrosine in breath condensate and exhaled NO in patients with mild asthma (r = 0.65, p < 0.05). We conclude that nitrotyrosine formation in exhaled breath condensates may be a marker of oxidative stress in airways of asthma. PMID- 11029331 TI - An investigation into causative factors in patients with bronchiectasis. AB - Bronchiectasis is a pathologic description of lung damage characterized by inflamed and dilated thick-walled bronchi. These findings may result from a number of possible causes and these may influence treatment and prognosis. The aim of this study was to determine causative factors in 150 adults with bronchiectasis (56 male, 94 female) identified using high-resolution computerized tomography. Relevant factors were identified in the clinical history; cystic fibrosis gene mutation analysis was performed; humoral immune defects were determined by measuring immunoglobulins, IgG subclasses and functional response to Pneumovax II vaccine; assessment was made of neutrophil function (respiratory burst, adhesion molecule expression, and chemotaxis); ciliary function was observed and those likely to have allergic bronchopulmonary aspergillosis (ABPA) were identified. Causes identified were: immune defects (12 cases), cystic fibrosis (4), Young's syndrome (5), ciliary dysfunction (3), aspiration (6), panbronchiolitis (1), congenital defect (1), ABPA (11), rheumatoid arthritis (4), and early childhood pneumonia, pertussis, or measles (44). Intensive investigation of this population of patients with bronchiectasis led to identification of one or more causative factor in 47% of cases. In 22 patients (15%), the cause identified had implications for prognosis and treatment. PMID- 11029332 TI - Effect of supine posture on respiratory mechanics in chronic left ventricular failure. AB - The mechanisms of orthopnea and the role of changes in respiratory mechanics in left ventricular failure (LVF) are poorly understood. We have measured total respiratory airflow resistance (Rrs) using forced oscillation in the sitting and supine positions in 10 patients with chronic LVF (NYHA II-III) shortly after recovery from acute LVF and in 10 matched control subjects (CON). Seated, the patients with LVF had small lung volumes but no evidence of airway obstruction (mean FEV(1)/FVC, 81%). Mean Rrs at 6 Hz was only slightly higher in LVF (3.4 cm H(2)O. L(-1). s) than in CON (2.6 cm H(2)O. L(-1). s). After 5 min supine, breathlessness in LVF increased. Despite much smaller mean falls in mid-tidal lung volume (MTLV) in LVF than in CON, the supine rise in Rrs was 80.5% in LVF and 37.6% in CON; mean increases in specific Rrs (SRrs = Rrs.MTLV) were 75.8% in LVF and 16.6% in CON (p 0.001). Five minutes after resuming the sitting position all values had reverted almost to the original sitting values. In 5 LVF patients, nebulized ipratropium, a muscarinic antagonist, only slightly attenuated the supine rise in SRrs. We conclude that patients with chronic LVF, who had little evidence of airways obstruction when seated, showed a large rise in airflow resistance after lying supine for 5 min. This cannot be attributed to reduction in lung volume when supine and no evidence was found of vagally-induced bronchoconstriction. Further experiments are required to establish the cause of the rapid supine rise in airflow resistance in LVF. PMID- 11029333 TI - Acute respiratory failure after lung volume reduction surgery. AB - In this study, we characterized patients who developed respiratory failure postoperatively after lung volume reduction surgery (LVRS). We retrospectively reviewed the records of 72 patients who underwent LVRS from February 1995 to February 1998, examining perioperative variables and complications. Twenty-one patients (29%) developed postoperative respiratory failure, five due to hypoxemia, nine due to hypercapnia, and seven secondary to hemodynamic instability. The hospital mortality was 33% among patients who developed respiratory failure. No preoperative clinical or physiologic variable (including percent ideal body weight, serum albumin, prednisone use, lung function, maximal O(2) uptake on exercise testing, 6-min walk distance, and hemodynamic parameters) was predictive of postoperative respiratory failure. Patients who developed respiratory failure were older (63 +/- 7 versus 57 +/- 8 yr, p = 0.01), had longer anesthesia time (188 +/- 96 versus 127 +/- 56 min, p = 0.001), had a higher incidence of coronary artery disease (40% versus 10%, p = 0.001) and performance of concomitant surgical procedures during the LVRS operation (40% versus 2%, p < 0.001) compared with those without respiratory failure. All patients who underwent simultaneous surgery, which were mostly for cardiac disease, developed respiratory failure. Risk factor analysis confirmed that older patients and those undergoing cardiac surgery combined with LVRS are at increased risk for postoperative respiratory failure. PMID- 11029334 TI - Antiasthmatic effects of mediator blockade versus topical corticosteroids in allergic rhinitis and asthma. AB - To compare the antiasthmatic efficacy of inflammatory mediator blockade versus topical corticosteroid therapy in patients with seasonal allergic rhinitis (SAR) and asthma, 14 patients were enrolled into a single-blind, double-dummy, placebo controlled crossover study comparing 2 wk therapy of (1) 400 microgram orally inhaled budesonide plus 200 microgram intranasal budesonide (BUD) or (2) 10 mg oral montelukast plus 10 mg oral cetirizine (ML + CZ). Before each treatment period, patients received 7 to 10 d placebo washout. All treatments were given once daily in the morning. Throughout the study, patients recorded the following domiciliary measures: peak expiratory flow (PEF), rescue inhaler requirement, asthma symptoms, and daily activity score. Laboratory measurements were made at trough of adenosine monophosphate (AMP) bronchial challenge and exhaled nitric oxide (NO). Compared with pooled placebo (PL), there were significant (p < 0.05) improvements in all domiciliary measures with both treatments (mean PEF [L/min] PL: 463; BUD: 478; ML + CZ: 483). For geometric mean AMP PC(20) (mg/ml), there was an improvement (p < 0.05), compared with PL (47), for ML + CZ (133) but not for BUD (51); whereas for NO (ppb) there was significant suppression with BUD (7.6) but not ML + CZ (11.5) compared with PL (13.6). In conclusion, both combined mediator blockade and combined topical corticosteroids are equally effective antiasthma therapy in patients with asthma and SAR. PMID- 11029335 TI - Exercise-induced bronchoconstriction does not cause eosinophilic airway inflammation or airway hyperresponsiveness in subjects with asthma. AB - The cysteinyl leukotrienes (LT) C(4), D(4), and E(4) may partially mediate eosinophilic airway inflammation in patients with asthma. High- intensity exercise by patients with asthma can result in exercise- induced bronchoconstriction, partly due to leukotriene production, but it is still debated whether this causes airway inflammation. Ten subjects completed a randomized, controlled study to examine the effects of exercise-induced bronchoconstriction on airway inflammatory cells. Subjects completed exercise challenge and methacholine challenge in random order separated by 1 wk. Spirometry was measured for 2 h after challenges, and airway responsiveness was measured the day before and 24 h after each challenge. Blood and sputum samples were obtained before, and 2, 4, 7, and 24 h after each challenge for measurement of inflammatory cells. Nine of the subjects inhaled allergen at least 3 wk before or 1 wk after the study. Sputum samples were collected before, 7 h, and 24 h after challenge. The maximum percentage fall in FEV(1) was 21.3 +/- 1.5% after exercise, 29.9 +/- 1.5% after methacholine, and 28.9+/-2.7% after allergen. Exercise had no effect on airway responsiveness or inflammatory cells measured in blood or sputum, unlike allergen inhalation, which resulted in significant airway hyperresponsiveness and increases in sputum eosinophils (p < 0.05). This study demonstrates that exercise-induced bronchoconstriction does not cause eosinophilic airway inflammation in subjects with asthma who develop airway inflammation with the same degree of allergen-induced bronchoconstriction. We conclude that exercise-induced bronchoconstriction does not cause airway inflammation or airway hyperresponsiveness. PMID- 11029336 TI - Airway hyperresponsiveness, inflammation, and subepithelial collagen deposition in recently diagnosed versus long-standing mild asthma. Influence of inhaled corticosteroids. AB - This study aimed at documenting airway inflammation and subepithelial collagen deposition in patients using only inhaled beta(2)-agonists with either recently diagnosed asthma (RDA: /= 13 yr, n = 16) and at the influence of an intense inhaled corticosteroid (ICS) treatment on these parameters, in relation to changes in airway responsiveness. Patients had a methacholine inhalation test and a bronchoscopy with bronchial biopsies before and after an 8-wk treatment with inhaled fluticasone propionate (FP), 1,000 microgram/day. Baseline FEV(1) (mean +/- SEM) was normal and similar in both groups (RDA: 98.1 +/- 2.7, LSA: 94.5 +/- 4.6%). Geometric mean methacholine PC(20) was lower in LSA than in RDA (0.44 versus 3.37 mg/ml) at baseline and improved similarly by 1.85 and 1.86 double concentrations with FP treatment. PC(20) normalized (>/= 16 mg/ml) in five patients with RDA and two patients with LSA. Baseline mean bronchial cell counts (per mm(2) connective tissue surface) for CD3(+), CD4(+), CD8(+), CD25(+), EG1(+), CD45ro(+), and AA1(+) cells were similar in both groups. With FP, EG1(+) (p < 0.001), EG2(+) (p = 0.018), and AA1(+) counts (p = 0.009) decreased significantly in both groups while CD45ro(+) (p = 0.02) counts decreased only in LSA. Baseline type 1 and type 3 collagen deposition underneath the basement membrane was similar in RDA and LSA and did not change significantly after FP. This study shows that recent compared to long standing mild asthma is associated with a similar degree of airway inflammation and subepithelial fibrosis, and a similar improvement in airway hyperresponsiveness after 8 wk on high-dose ICS. It also indicates that once asthma becomes symptomatic, airway responsiveness cannot normalize in most subjects over such a time period, even with a high dose of ICS. PMID- 11029337 TI - Risk of tuberculosis infection and tuberculous meningitis after discontinuation of Bacillus Calmette-Guerin in Beijing. AB - In Beijing, the notification rate of smear-positive tuberculosis (TB) has been below 20 per 100,000 since 1986, and continues to decline. To accurately measure the risk of TB infection in a population in which the results of tuberculin skin testing were not confounded by vaccination with Bacillus Calmette-Guerin (BCG), BCG vaccination at birth was discontinued from 1988 in Shun-yi County. In 1995, the prevalence of TB infection among 12,836 primary school children aged 6 to 7 yr and without BCG scars was 1.4%, giving an estimated annual risk of infection of 0.19% (95% confidence interval: 0.16 to 0.22%). The prevalence of TB infection in children aged 5 to 9 yr in Beijing in 1950 was 46%. The number of cases of tuberculous meningitis did not increase after discontinuation of BCG. We conclude that discontinuation of BCG had no detectable harmful effects, and that control of TB in Beijing has markedly reduced the prevalence of TB infection since 1950. PMID- 11029338 TI - Screening for bronchial hyperresponsiveness using methacholine and adenosine monophosphate. Relationship to asthma severity and beta(2)-receptor genotype. AB - Bronchial hyperresponsiveness (BHR) is a key feature of asthma and may be measured by direct methacholine challenge or indirect adenosine monophosphate (AMP) challenge. We performed a retrospective analysis of our database (n = 487) of patients with asthma with the aim first, to compare methacholine and AMP challenge as screening tools, and second, to identify any relationships between BHR and disease severity markers or beta(2)-adrenoceptor genotype. Of these subjects, 258 had a methacholine challenge, 259 an AMP challenge and 185 both. Of subjects having both, 140 (76%) were methacholine responsive with PD(20) < 500 microgram (PC(20) < 5 mg/ml) and 92 (50%) were AMP responsive with PC(20) < 200 mg/ ml. For those who were AMP unresponsive 57% were methacholine responsive, whereas for the methacholine nonresponders 11% were AMP responsive. Methacholine (but not AMP)-responsive patients had a significantly (p < 0.05) lower % predicted FEV(1) and FEF(25-75) and higher inhaled corticosteroid dose than unresponsive patients. Finally, subjects with a glycine allele at codon 16 had significantly (p < 0.05) increased BHR to methacholine but not AMP. Our results suggest that methacholine is a more appropriate screening tool for BHR than AMP as it was more sensitive in our population and was also related to asthma severity. In addition, we have demonstrated an association between the glycine allele (codon 16) and increased BHR to methacholine. PMID- 11029339 TI - Accuracy and utility of commercially available amplification and serologic tests for the diagnosis of minimal pulmonary tuberculosis. AB - Diagnosis of patients with minimal active tuberculosis (TB) is difficult, as there is no single test with high sensitivity and specificity. The yield and clinical utility of a combination of diagnostic tests were prospectively studied among 500 consecutive patients referred for sputum induction for diagnosis of possible active TB. Patients underwent sputum induction, chest X-ray, tuberculin testing, and had blood drawn for serologic testing (Detect-TB test; Biochem ImmunoSystems). Sputum was examined with fluorescent microscopy and PCR (Amplicor MTB-Roche) and cultured for mycobacteria using liquid (BACTEC) and solid media. For the diagnosis of the 60 cases of active TB, sensitivity and specificity, respectively, of the following diagnostic tests were mycobacterial culture, 73% and 100%; PCR, 42% and 100%; chest X-ray, 67-77% and 66-76%; tuberculin testing, 94% and 20%; and serology, 33% and 87%. After consideration of PCR and radiographic and clinical characteristics, a positive serologic test was independantly associated with diagnosis of active disease (adjusted odds of disease if positive, 2.6; 95% confidence limits, 1.1,6.1). No currently available test has sensitivity and specificity high enough for the accurate diagnosis of minimal pulmonary TB. Utilization of a combination of tests, together with consideration of key clinical characteristics, could improve diagnostic accuracy. PMID- 11029340 TI - Measuring asthma control. Clinic questionnaire or daily diary? AB - Daily symptom, peak expiratory flow rate (PEFR), and medication diaries are often used in clinical trials of treatments for asthma on the assumption that they provide a better estimate of clinical status than does a questionnaire completed in the clinic. We conducted a study with the aim of comparing the measurement properties of the clinic-completed Asthma Control Questionnaire with those of the Asthma Control Diary. The diary is composed of questions and response options almost identical to those of the questionnaire, but uses PEFR instead of FEV(1) as the measure of airway caliber. In an observational study, 50 adults with symptomatic asthma attended a McMaster University asthma clinic at 0, 1, 5, and 9 wk to complete the Asthma Control Questionnaire and other measures of asthma status. For 1 wk before each follow-up visit, patients completed the Asthma Control Diary every morning and evening. Concordance between the questionnaire and diary was high (intraclass correlation coefficient [ICC] = 0.87). Both reliability (ICC: questionnaire = 0.90; diary = 0.86) and responsiveness (responsiveness index: questionnaire = 1.06; diary = 0.90; p = 0.005) were better with the questionnaire than with the diary. Correlations between the two instruments and other measures of clinical asthma status were similar and close to a priori predictions. Both the Asthma Control Questionnaire and the Asthma Control Diary are valid instruments for measuring asthma control, but the questionnaire has slightly better discriminative and evaluative measurement properties than does the diary. PMID- 11029341 TI - Randomized controlled trial of domiciliary noninvasive positive pressure ventilation and physical training in severe chronic obstructive pulmonary disease. AB - The addition of noninvasive positive pressure ventilation (NPPV) to an exercise training (ET) program in severe chronic obstructive pulmonary disease (COPD) may produce greater benefits in exercise tolerance and quality of life than after training alone. Forty-five patients with severe stable COPD-mean (SD) FEV(1) 0.96 (0.31) L, Pa(O(2)) 65.4 (9.07) mm Hg, Pa(CO(2)) 45.6 (7.89) mm Hg-were randomized to domiciliary NPPV + ET (n = 23) or ET alone (n = 22). Exercise capacity and health status were assessed at baseline and after an 8-wk training program. There was a significant improvement in mean shuttle walk test (SWT) in the NPPV + ET group: from 169 (112) to 269 (124) m (p = 0.001), compared with the ET group: 205 (100) to 233 (123) m (p = 0.19); mean difference (95% confidence interval [CI]): 72 (12.9 to 131) m. Repeated measures analysis of variance (ANOVA) showed that the differences between the two groups became evident only in the final 4 wk of the training program with a mean end study difference (95% 1CI) of 65.8 (17.1 to 114) m. There was a significant improvement in the Chronic Respiratory Disease Questionnaire (CRDQ) of mean (SD) 24.0 (17.4) (p = < 0.001) in the NPPV + ET group and 11.8 (15.8) (p = 0.003) points in the ET group; mean difference: 12.3 (1.19 to 23.4). Only the NPPV + ET group demonstrated a significant improvement in arterial oxygenation; mean difference: 3.70 mm Hg (0.37 to 7.27). This study suggests that domiciliary NPPV can be used successfully to augment the effects of rehabilitation in severe COPD. PMID- 11029342 TI - Ozone, but not nitrogen dioxide, exposure decreases glutathione peroxidases in epithelial lining fluid of human lung. AB - Antioxidants, such as glutathione peroxidases (GPxs), in epithelial lining fluid (ELF) protect against health effects of oxidant pollutants, which includes O(3) or NO(2). We hypothesized that GPxs concentration in ELF is responsive to O(3) or NO(2) exposure. Subjects underwent two 4-h exposures to O(3) (0.22 ppm) and one to air. In another experiment, subjects underwent 3-h exposures to air and NO(2) (0.6 and 1.5 ppm). Bronchoalveolar lavage (BAL) was performed immediately or 18 h after O(3) exposure and 3.5 h after each NO(2) exposure. GPx activity and extracellular GPx (eGPx) protein concentrations were determined in ELF, and their relationships to markers of lung function, inflammation, and epithelial permeability were examined. Although the total amounts were not changed, basal (air) GPx activity (223.6 +/- 24.4 mU/ml), basal eGPx protein concentration (2.62 +/- 0.25 microg/ml), and basal ELF dilution factor (152.3 +/- 8.4) decreased 40% immediately after O(3) exposure and remained 30% decreased 18 h after exposure (p = 0.0001). No effect of NO(2) exposure on GPxs concentration was detected. There was an inverse correlation between baseline ELF eGPx protein concentration and the change in PMN 18 h after O(3) exposure (p = 0.04). Thus, O(3), a strong oxidant, decreases both GPx activity and eGPx protein in ELF, whereas NO(2), a weaker oxidant, does not. eGPx in ELF may protect against O(3)-induced airway inflammation. PMID- 11029343 TI - (1-->3)-beta-D-glucan and endotoxin in house dust and peak flow variability in children. AB - House dust-associated bacterial endotoxins have been shown to be associated with asthma severity, and a similar role has been suggested for fungal (1-->3)-beta-D glucans. In this study the relation between these agents and peak expiratory flow (PEF) variability was investigated in 148 children 7 to 11 yr of age of whom 50% had self- or parent-reported chronic respiratory symptoms. All children self monitored twice daily their PEF for a period of 16 wk. Dust samples were collected from mattresses and from living room and bedroom floors, and endotoxin and (1-->3)-beta-D-glucan were measured in dust extracts. The relations with mean daily PEF variability (Ampl%mean) were investigated by linear regression analysis, adjusting for dust mite allergen levels, presence of pets, and type of floor cover. In unadjusted analyses the levels of both endotoxin and (1--> 3) beta-D-glucan per square meter of living room floor were significantly associated with PEF-variability (but not when expressed per gram of sampled dust), particularly in atopic children with asthma symptoms. Adjusted analyses showed the same association for (1--> 3)-beta-D-glucan but not for endotoxin. Although no associations were found with microbial agent levels in bedroom floor or mattress dust, high levels of (1-->3)-beta-D-glucan in living room floor dust apparently increase PEF variability in asthmatic children. PMID- 11029344 TI - Balance of matrix metalloprotease-9 and tissue inhibitor of metalloprotease-1 from alveolar macrophages in cigarette smokers. Regulation by interleukin-10. AB - An imbalance between proteases and antiproteases may play a role in emphysema, which is characterized by increased degradation of extracellular matrix, and in airway remodeling in chronic bronchitis and asthma, in which there is increased collagen deposition. We assessed the effect of smoking on release of matrix metalloprotease-9 (MMP-9) and of its inhibitor, tissue inhibitor of metalloprotease-1 (TIMP-1), from alveolar macrophages, and determined the effects of proinflammatory (interleukin [IL]-1beta and lipopolysaccharide [LPS]) and antiinflammatory (IL-10) stimuli on the release of MMP-9 and TIMP-1. We performed bronchoalveolar lavage in 11 smokers and 11 nonsmokers, and cultured airway macrophages in the presence of control medium, IL-1beta, and LPS. Airway macrophages from smokers released greater amounts of MMP-9 and TIMP-1 at baseline and in response to IL-1beta and LPS than did those of nonsmokers. Airway macrophages from smokers produced more TNF-alpha and IL-10. IL-10 increased TIMP 1 release without modifying that of MMP-9, leading to a decrease in the MMP-9 to TIMP-1 ratio. Anti-IL-10 antibody had no effect on MMP-9 production induced by LPS. We conclude that the release of proteases and antiproteases by airway macrophages is increased in cigarette smokers, and can be regulated by exogenous IL-10. PMID- 11029345 TI - Tromethamine buffer modifies the depressant effect of permissive hypercapnia on myocardial contractility in patients with acute respiratory distress syndrome. AB - In patients with acute respiratory distress syndrome (ARDS), permissive hypercapnia is a strategy to decrease airway pressures to prevent ventilator induced lung damage by lowering tidal volumes and tolerating higher arterial carbon dioxide tension. However, in experimental studies hypercapnia impairs myocardial contractility and hemodynamic function. We investigated the effect of short-term permissive hypercapnia on myocardial contractility and hemodynamics in patients with ARDS. We hypothesized that the administration of tromethamine (THAM), a buffer which does not increase carbon dioxide production, would modify these changes. In 12 patients with ARDS, permissive hypercapnia was implemented for 2 h with a target Pa(CO(2))of 80 mm Hg. Patients were randomized to have respiratory acidosis corrected by THAM (pH-corrected group), or not corrected (pH uncorrected group). Hemodynamic responses were measured, and transesophageal echocardiography (TEE) was used to determine myocardial contractility. Permissive hypercapnia resulted in significant decreases in systemic vascular resistance (SVR) and increases in cardiac output (Q). Myocardial contractility decreased in both groups but significantly less in the pH-corrected group (approximately 10%) than in the pH-uncorrected group (approximately 18%, p < 0.05). Mean arterial pressure decreased and mean pulmonary arterial pressure increased significantly only in the pH-uncorrected group. All values returned to baseline conditions 1 h after permissive hypercapnia was terminated. Our study demonstrates a reversible depression of myocardial contractility and hemodynamic alterations during rapid permissive hypercapnia which were attenuated by buffering with THAM. This may have applicability to the clinical strategy of permissive hypercapnia and allow the benefit of decreased airway pressures to be realized while minimizing the adverse hemodynamic effects of hypercapnic acidosis. PMID- 11029346 TI - Acid instillation enhances the inflammatory response to subsequent lipopolysaccharide challenge in rats. AB - Aspiration of gastric contents is one of leading causes of the acute respiratory distress syndrome (ARDS). The pathogenesis of acid aspiration-induced acute lung injury is well understood. Less clear is why patients who have suffered acid aspiration are susceptible to ARDS. We studied the effects of acid instillation on the inflammatory response to subsequent lipopolysaccharide (LPS) challenge in rats. Instillation of acid into the right lung worsened the pathology induced by LPS that was administered 24 h after acid instillation. This included worsened oxygenation, increased pulmonary edema, increased production of tumor necrosis factor-alpha (TNF-alpha) and cytokine-induced neutrophil chemoattractant, neutrophil accumulation and mobilization to the alveolar spaces, and nitric oxide (NO) production. Of interest, neutrophil mobilization, NO production, and protein permeability were also magnified in the left lung. These effects were attenuated by administration of the protein tyrosine kinase (PTK) inhibitors genistein and tyrphostin AG556. These data suggest that acid instillation primes the rat to enhance the inflammatory response to subsequent endotoxin challenge and that at least part of the augmented inflammatory response depends on PTK. PMID- 11029347 TI - Accuracy of hemodynamic measurements during partial liquid ventilation with perflubron. AB - Patients undergoing partial liquid ventilation (PLV) are often monitored with pulmonary artery catheters and receive positive end-expiratory pressure (PEEP). PEEP can dissociate wedge pressure (Pcw) from transmural left atrial pressure (Platm) by elevating pleural pressure and can dissociate Pcw from Pla by elevating alveolar pressure, PLV, like PEEP, also elevates pleural and alveolar pressures. However, the artifacts PLV may cause in measured vascular pressures are unknown. In 6 anesthetized, paralyzed healthy adult sheep, we compared effects of gas ventilation (GV) and PLV with 10 and 30 ml/kg perflubron on pericardial pressure (Pperi), Pcw, Pla, thermodilution cardiac output, and pulmonary artery flow measured with a doppler probe. PEEP was applied from 0-15 mm Hg during GV and PLV. PLV changed pericardial pressure or cardiac output minimally (at PEEP(0), GV: Pperi = -1.7 +/- 0.6 mm Hg, CO = 3. 2 +/- 0.1 L/m; 10 ml/kg perflubron: Pperi = -1.3 +/- 0.6 mm Hg, CO = 3.4 +/- 0.2 L/m; 30 ml/kg perflubron: Pperi = -1.6 +/- 0.7 mm Hg, CO = 3.4 +/- 0.2 L/m; all mean +/- SEM). On PEEP, Pcw agreed with Pla and Platm as well or better during PLV as during gas ventilation. Cardiac output by thermodilution and probe agreed equally well under all conditions. We conclude that hemodynamic values are as accurate during PLV as during gas ventilation. PMID- 11029348 TI - Attenuation of the allergen-induced late asthmatic reaction by cyclosporin A is associated with inhibition of bronchial eosinophils, interleukin-5, granulocyte macrophage colony-stimulating factor, and eotaxin. AB - The allergen-induced late asthmatic reaction (LAR) is associated with increases in bronchial eosinophils and basophils as well as upregulation of several eosinophil active cytokines and C-C chemokines. Cyclosporin A (CsA) was previously shown to inhibit the LAR, but not the early asthmatic reaction (EAR), and this was associated with a decrease in blood eosinophils. For these reasons, we determined whether CsA inhibited the allergen-induced increases in bronchial eosinophils, basophils, eotaxin, interleukin-5 (IL-5), and granulocyte macrophage colony-stimulating factor (GM-CSF). Subjects with a demonstrable LAR underwent bronchoscopy with biopsy and bronchoalveolar lavage (BAL) at baseline and then were randomly allocated to receive either CsA (n = 13) or placebo (n = 11) before challenge. A second bronchoscopy was performed 24 h later. The LAR, but not the EAR, was significantly attenuated in the CsA group compared with placebo (p < 0.05). CsA significantly inhibited the allergen-induced increases in IL-5 (p = 0.02) and GM-CSF (p = 0. 0028) in mRNA+ cells in BAL, and in a mAB against human activated eosinophils (EG2+) (p = 0.0227). We conclude that inhibition of the LAR by CsA may be related to its inhibitory effects on eosinophil-associated cytokines and chemokines. The beneficial effect of CsA in asthma may also be the result of inhibition of eosinophil accumulation. PMID- 11029350 TI - Quality of life in allergic rhinitis and asthma. A population-based study of young adults. AB - Quality of life has been found to be impaired both in patients with asthma and in patients with allergic rhinitis, but the relative burden of these diseases has not been investigated. We analyzed answers to the SF-36 questionnaire from 850 subjects recruited in two French centers participating in the European Community Respiratory Health Survey, a population-based study of young adults. Both asthma and allergic rhinitis were associated with an impairment in quality of life. However, 78% of asthmatics also had allergic rhinitis. Subjects with allergic rhinitis but not asthma (n = 240) were more likely than subjects with neither asthma nor rhinitis (n = 349) to report problems with social activities, difficulties with daily activities as a result of emotional problems, and poorer mental well-being. Patients with both asthma and allergic rhinitis (n = 76) experienced more physical limitations than patients with allergic rhinitis alone, but no difference was found between these two groups for concepts related to social/mental health. As asthma was not found to further impair the quality of life in subjects with allergic rhinitis for concepts related to mental disability and well-being, and as subjects with asthma often also suffer from allergic rhinitis, further studies on quality of life in asthma should ensure that the impairment in quality of life attributed to asthma could not result from concomitant allergic rhinitis. PMID- 11029349 TI - Association between air pollution and lung function growth in southern California children. AB - Average growth of lung function over a 4-yr period, in three cohorts of southern California children who were in the fourth, seventh, or tenth grade in 1993, was modeled as a function of average exposure to ambient air pollutants. In the fourth-grade cohort, significant deficits in growth of lung function (FEV(1), FVC, maximal midexpiratory flow [MMEF], and FEF(75)) were associated with exposure to particles with aerodynamic diameter less than 10 micrometer (PM(10)), PM(2.5), PM(10)-PM(2.5), NO(2), and inorganic acid vapor (p < 0.05). No significant associations were observed with ozone. The estimated growth rate for children in the most polluted of the communities as compared with the least polluted was predicted to result in a cumulative reduction of 3.4% in FEV(1) and 5.0% in MMEF over the 4-yr study period. The estimated deficits were generally larger for children spending more time outdoors. In the seventh- and tenth-grade cohorts, the estimated pollutant effects were also negative for most lung function measures, but sample sizes were lower in these groups and none achieved statistical significance. The results suggest that significant negative effects on lung function growth in children occur at current ambient concentrations of particles, NO(2), and inorganic acid vapor. PMID- 11029351 TI - Airway and respiratory tissue mechanics in normal infants. AB - Low-frequency respiratory impedance (Zrs) was measured by applying a forcing signal, between 0.5 and 21 Hz at a transrespiratory pressure of 20 cm H(2)O, in a cross-sectional study of 37 normal infants. A model containing an airway resistance (Raw) and inertance (Iaw) and a tissue damping (G) and elastance (H) was fitted to the individual Zrs. Forced expiratory volume in 0.5 second (FEV(0.5)) was determined using the raised volume rapid thoracic compression technique. Multivariate regression analysis was used to analyze the relationships between the lung function parameters and length. Both airway and tissue parameters showed a decreasing quadratic relationship with increasing length. FEV(0.5) showed an increasing cubic relationship with length. A family history of asthma was found to have a negative effect on Raw, H, and FEV(0.5). PMID- 11029352 TI - A clinical index to define risk of asthma in young children with recurrent wheezing. AB - Because most cases of asthma begin during the first years of life, identification of young children at high risk of developing the disease is an important public health priority. We used data from the Tucson Children's Respiratory Study to develop two indices for the prediction of asthma. A stringent index included frequent wheezing during the first 3 yr of life and either one major risk factor (parental history of asthma or eczema) or two of three minor risk factors (eosinophilia, wheezing without colds, and allergic rhinitis). A loose index required any wheezing during the first 3 yr of life plus the same combination of risk factors described previously. Children with a positive loose index were 2.6 to 5.5 times more likely to have active asthma between ages 6 and 13 than children with a negative loose index. Risk of having subsequent asthma increased to 4.3 to 9.8 times when a stringent index was used. We found that 59% of children with a positive loose index and 76% of those with a positive stringent index had active asthma in at least one survey during the school years. Over 95% of children with a negative stringent index never had active asthma between ages 6 and 13. We conclude that the subsequent development of asthma can be predicted with reasonable accuracy using simple, clinically based parameters. PMID- 11029353 TI - Habitually sleepy drivers have a high frequency of automobile crashes associated with respiratory disorders during sleep. AB - Sleepiness is a common cause of traffic crashes with a cost of billions of dollars per year. A recent study has found that 2 to 3% of drivers are habitually sleepy while driving. However, there has not been a controlled study to define the characteristics, driving performance, or automobile crash rate of habitually sleepy drivers. The prevalence of respiratory disorders during sleep, and whether these respiratory disorders contribute to the increased automobile crash frequency, is unknown in habitually sleepy drivers. We interviewed 4,002 randomly selected drivers to define the prevalence of drivers who are habitually sleepy while driving. We studied the habitually sleepy drivers and an age- and sex matched control group of drivers. These studies included reporting of daytime sleepiness, automobile crashes, driving performance and sleep studies. Of the 4, 002 drivers interviewed, 145 (3.6%, confidence interval [CI] = 3.1 to 4.3) were habitually sleepy while driving. The habitually sleepy drivers reported a significantly higher frequency of auto crashes than control subjects (the adjusted odds ratio [OR] was 13.3, CI = 4. 1 to 43). The habitually sleepy drivers had a significantly higher prevalence of respiratory sleep disorders than control subjects. For a total respiratory events index (apneas, hypopneas, and other respiratory effort-related arousals) >/= 15 the adjusted OR was 6.0, CI = 1.1 to 32. In the habitually sleepy drivers group, the frequency of sleep apnea (apnea-hypopnea index) between subjects with or without auto crashes was not statistically different. However, if we consider total respiratory events index, this frequency of respiratory sleep disorders was significantly higher in subjects with automobile crashes (the adjusted OR for a total respiratory event index >/= 15 was 8.5, CI = 1.2 to 59). Habitually sleepy drivers are a large group of drivers (1 of 30 drivers) who are involved in several fold more automobile crashes than control subjects. As these excess auto crashes can be explained in part by the presence of respiratory disorders during sleep, which are treatable, many automobile crashes in these sleepy drivers may be preventable. Our findings suggest that asking about excessive sleepiness while driving may better predict which subjects with breathing disorders during sleep have crashes than asking about overall sleepiness. PMID- 11029354 TI - A noninvasive diagnostic strategy including spiral computed tomography in patients with suspected pulmonary embolism. AB - We conducted a study to evaluate a noninvasive strategy including spiral computed tomography (CT) in patients with suspected pulmonary embolism (PE). We systematically performed spiral CT, ventilation/perfusion lung scanning, and D dimer (DD) measurement (VIDAS test), and in some cases (with a normal CT with nondiagnostic lung scan and increased DD) performed venous ultrasonography (US) on 247 consecutive patients with clinically suspected PE in our hospital. Patients in whom PE was deemed absent were not given anticoagulants. All patients were followed for 3 mo. The prevalence of PE in the 228 patients who could be evaluated was 42% (96 of 228). PE was confirmed by spiral CT in 73% of the patients, by a high-probability lung scan in 4%, and by findings on US in 23%. PE was ruled out by a normal lung scan in 14% of the patients, by a normal DD concentration (< 500 ng/ml) in 31%, by an obvious differential diagnosis on spiral CT in 18%, by a similar prior lung scan in 11%, and by the combination of normal spiral CT findings, a nondiagnostic lung scan, a DD concentration > 500 ng/ml, and normal US in 26%. Pulmonary angiography was performed in only two patients, both of whom had a normal spiral CT scan and a high-probability lung scan, and was normal. The 3-mo risk of thromboembolism in patients not given anticoagulants, based on the results of the diagnostic protocol, was 1.7% (95% confidence interval: 1.5 to 2.3%). There were no deaths. The noninvasive strategy of combining spiral CT, lung scanning, DD measurement, and in some cases US, in patients with suspected PE yielded a definite diagnosis in 99% of patients, and appeared to be safe. PMID- 11029355 TI - Does size matter? Utility of size of tuberculin reactions for the diagnosis of mycobacterial disease. AB - It is a common belief that larger tuberculin reactions are more serious, and more likely to indicate patients with active tuberculosis (TB) or at high risk of disease in the future. Among 182 close contacts, and 502 patients suspected of possible active TB, 529 underwent tuberculin skin testing (TST) and 605 had a chest radiograph. Final diagnoses, based on all available clinical, microbiological, histological, and radiographic information, were active TB, 68; inactive TB, 274; nontuberculous mycobacterial disease, 14; conditions associated with anergy, 36; no detectable abnormality (except a positive TST) or condition unrelated to TB, 213; and negative TST, no further evaluation, 79. Among these patients, TST of 5 mm or larger was significantly more likely to indicate active or inactive TB (p < 0.001). However, among patients with TST of 5 mm or greater, the size and frequency distribution of tuberculin reactions were not different between subjects with different diagnoses, nor between subjects with different types or extent of radiographic findings. As well, TST reactions were no different in 121 subjects with or 176 subjects without a history of BCG vaccination. In close contacts or patients suspected of active TB, reactions less than 5 mm indicated lower likelihood of active or inactive disease, but above that threshold, size of tuberculin reaction did not matter. PMID- 11029356 TI - Duration of asthma and physiologic outcomes in elderly nonsmokers. AB - Airway and alveolar inflammation have been described in asthma. Prolonged inflammation may lead to airway remodeling, which can result in physiologic abnormalities. Elderly lifetime nonsmokers are an ideal population in which to examine the consequences of longstanding asthma. To test the hypothesis that airflow limitation and hyperinflation are associated with the duration of asthma, we evaluated airflow and lung volumes in a cohort of elderly asthmatic individuals. All subjects were > 60 yr of age and were lifetime nonsmokers (n = 75). Patients with asthma of long duration (LDA; n = 38) had asthma for >/= 26 yr (median = 40.0 yr); patients with asthma of short duration (SDA; n = 37) had asthma for < 26 yr (median = 9 yr). Patients with LDA had a significantly lower FEV(1)% predicted than did those with SDA (59.5 +/- 2.6% versus 73.8 +/- 3.1% [mean +/- SEM], respectively; p < 0.007). Regression analysis demonstrated that duration of asthma was inversely associated with FEV(1)% predicted (r = 0.264, p < 0.03). After bronchodilator administration, the patients with LDA continued to show airflow obstruction (FEV(1)% predicted = 65.4 +/- 2.9). Only 18% of patients with LDA attained a normal postbronchodilator FEV(1), whereas 50% of those with SDA were able to do so (p < 0.003). The FRC% predicted was significantly higher in subjects with LDA than in those with SDA (142.9 +/- 5.6 versus 124.1 +/- 4.4, respectively, p < 0.01). Multiple regression analysis revealed an association between FRC and duration of asthma that was independent of the degree of airflow limitation. These data suggest that the duration of asthma is associated with the degree of airflow limitation and hyperinflation. Moreover, these abnormalities can become irreversible over time, and may reflect distal airway and/or parenchymal changes as well as proximal airway remodeling. PMID- 11029357 TI - Endothelial nitric oxide synthase function in pig lung after chronic pulmonary artery obstruction. AB - Because long-term pulmonary artery (PA) obstruction is associated with expansion of the systemic blood supply to the lung, chronic ischemia may not occur, and endothelium nitric oxide synthase (eNOS) function may be preserved in postobstructive pulmonary arteries. To test this hypothesis, we studied piglets 2 d or 5 wk after left PA ligation or a sham operation. We measured left lung ATP and lactate lung concentrations; calcium-dependent and calcium-independent NOS activities and eNOS protein; and left PA relaxations in response to acetylcholine, calcium ionophore, and sodium nitroprusside. Decreases in ATP and increases in lactate concentrations were significantly attenuated after 5 wk PA occlusion (p < 0.05 versus sham and 2-d ligation). Compared with sham and 2-d PA occlusion, calcium-dependent NOS activity and eNOS protein were lower in the long term PA occlusion group. Calcium-independent NOS activity was unchanged. Acetylcholine and calcium ionophore relaxations were impaired after 5 wk, whereas only acetylcholine relaxation was impaired after 2-d PA occlusion. Relaxation to sodium nitroprusside remained unchanged. In conclusion, despite relative conservation of lung energy metabolism, prolonged PA occlusion decreased eNOS function and protein in postobstructive pulmonary arteries. PMID- 11029358 TI - Role of CO diffusing capacity during exercise in the preoperative evaluation for lung resection. AB - We conducted a prospective study to evaluate whether lack of an adequate increase in diffusing capacity for carbon monoxide (DL(CO)) during exercise is associated with a greater postoperative complication rate after lung resection. We used the three-equation method (3EQ-DL(CO)), a modification of the single breath DL(CO) technique to determine DL(CO) during exercise in 57 patients undergoing lung resection at Vancouver General Hospital from October 1998 to May 1999. 3EQ-DL(CO) was determined during steady-state exercise at 35% and 70% of the maximal workload reached in a progressive exercise test. Maximal oxygen uptake (VO(2)max), DL(CO) at rest, and the increase in DL(CO) during exercise were compared in relation to postoperative complications. Patients with complications had lower resting values of DL(CO) (R-DL(CO)), a smaller increase in DL(CO) from rest to 70% of maximal workload expressed as a percent of the predicted DL(CO) at rest ([70% - R]-DL(CO)%), and a lower VO(2)max than did patients without complications. Results suggested that (70% - R)-DL(CO)% was the best preoperative predictor of postoperative complications; a cutoff limit of 10% was the best index to identify complications, yielding a complication rate of 100% in patients with (70% - R)-DL(CO)% < 10% as compared with a complication rate of 10% in patients with (70% - R)-DL(CO)% >/= 10% (sensitivity = 78%, specificity = 100%). Patients who do not increase their DL(CO) sufficiently during exercise ([70% - R] DL(CO)% < 10%) have a greater complication rate after lung resection. PMID- 11029360 TI - Elevation of exhaled ethane concentration in asthma. AB - Ethane is a product of lipid peroxidation as a result of oxidative stress and can be detected in the exhaled air. Oxidative stress plays a role in the pathogenesis of asthma. We measured exhaled ethane in 26 asthmatic subjects (mean age +/- SEM, 38 +/- 8 yr; 15 male, FEV(1) 60 +/- 4%) and compared it with exhaled nitric oxide (NO) measured by chemiluminescence, a noninvasive marker of oxidative stress and inflammation. Exhaled ethane was collected during a flow- and pressure-controlled exhalation into a reservoir discarding dead space air contaminated with ambient air. A sample of the expired air was analyzed by chromatography. Exhaled ethane levels were elevated in asthma patients not receiving steroid (n = 12, 2.06 +/- 0.30 ppb) compared with steroid-treated patients (n = 14, 0.79 +/- 0.10 ppb, p < 0.01) and to 14 nonsmoking control subjects (0.88 +/- 0.09 ppb, p < 0.05). In patients not receiving steroid treatment there was a positive correlation between exhaled ethane and NO (r = 0.55, p < 0.05) and air trapping assessed by the ratio of residual volume to total lung capacity (RV/ TLC) (r = 0.60, p < 0.05). In addition, untreated patients with FEV(1) < 60% predicted value had higher concentrations of ethane (2.86 +/- 0.37 ppb) compared with less obstructed patients (FEV(1) > 60%, 1.26 +/- 0.12 ppb, p < 0.05). NO concentrations were higher in patients not on steroid treatment (14.7 +/- 1.7 ppb) than in steroid treated patients (8.6 +/- 0.5 ppb, p < 0.05). Exhaled ethane is elevated in asthma, reduced in steroid-treated patients, and correlates with NO and airway obstruction. It may be a useful noninvasive marker of oxidative stress. PMID- 11029359 TI - Prospective randomized comparison of thoracoscopic talc poudrage under local anesthesia versus bleomycin instillation for pleurodesis in malignant pleural effusions. AB - Induction of pleurodesis offers benefit for patients with metastatic tumors and symptomatic malignant pleural effusions, but the best method for achieving this is still unknown. In this prospective, randomized comparison of two well established pleurodesis procedures, 36 patients with malignant pleural effusions, expanded lungs after drainage, and expected survival of > 1 mo received either bleomycin instillation (60E) via a small-bore thoracostomy tube or thoracoscopic talc poudrage (5 g) under local anesthesia. Efficacy, safety, and cost could be evaluated for 32 treatments (17 bleomycin, 15 talc) in 31 patients. Recurrence rates of effusion with bleomycin and talc poudrage after 30 d were 41% and 13% (p = 0.12), respectively, those after 90 d were 59% and 13%, respectively (p = 0.01), and those after 180 d were 65% and 13% (p = 0.005), respectively. Neither procedure showed any major adverse effect, and both were equally well tolerated. Cost estimation favored thoracoscopic talc poudrage, both for the initial hospitalization and with regard to recurrences. In conclusion, thoracoscopic talc pleurodesis under local anesthesia is superior to bleomycin instillation for pleurodesis in cases of malignant pleural effusion. PMID- 11029361 TI - Prolonged airway activity and improved selectivity of budesonide possibly due to esterification. AB - We addressed the question of whether the prolonged local retention of the glucocorticoid (GC) budesonide (BUD) within airway tissue, due to reversible fatty acid esterification, is associated with protracted topical anti inflammatory activity and improved airway selectivity, when compared with fluticasone propionate (FP). BUD or FP at 25 nmol/kg was administered intratracheally or subcutaneously to adrenalectomized rats, followed by lipopolysaccharide (LPS) intratracheal instillation. The trachea and main bronchi were lavaged 6 h after LPS, and tumor necrosis factor-alpha (TNF-alpha) concentration and cell number in the lavage fluid were measured. Instilled 1 h before LPS, both GCs reduced TNF-alpha by 70% (p < 0.05) and mononuclear cells by 55% (p < 0.01), with no reduction in neutrophils. Instilled 6 h before LPS, a significant reduction of TNF-alpha (59%, p < 0.02) and mononuclear cells (47%, p < 0.05) was achieved only with BUD. After subcutaneous administration, no significant effects were observed. BUD did not exert higher systemic activity than FP, measured as plasma corticosterone suppression. In conclusion, BUD exerted a more prolonged topical anti-inflammatory activity, and a higher airway selectivity than FP, possibly because of its reversible fatty acid esterification within airway tissue. This may contribute to the high efficacy and safety of BUD in asthma, even with once-daily inhalation. PMID- 11029362 TI - Utility of the antibody response to a conjugated Haemophilus influenzae type B vaccine for diagnosis of primary humoral immunodeficiency. AB - Antibody response to an Haemophilus influenzae type b (Hib)-conjugated vaccine was studied in 59 healthy adults (mean age: 32 yr) and 22 patients with humoral immunodeficiencies (mean age: 32 yr) to determine its usefulness in the diagnosis of defective antibody formation. Twenty of the healthy adults and nine of the patients were also immunized with a pneumococcal vaccine. Serum specific antibodies were measured by ELISA. Adequate response to both vaccines was defined using the lower limit of the two-tailed 90% probability interval of postimmunization specific IgG of the healthy adults. By using this cutoff, responders were considered to be those with an absolute increase in anti-Hib IgG titers higher than 2.28 microgram/ml, and in anti-Streptococcus pneumoniae IgG higher than 395 arbitrary units/ml. With these criteria, 85% (50 of 59) of the healthy adults responded with anti-Hib IgG and 75% (15 of 20) with anti pneumococcal IgG. All healthy adults receiving both vaccines responded to at least one. None of the patients with humoral immunodeficiencies responded to either vaccine. Evaluation of the antibody response to both the Hib and pneumococcal vaccines may facilitate the diagnosis of humoral immunodeficiency and selection of patients to receive immunoglobulin therapy. PMID- 11029363 TI - CXC chemokines IP-10 and mig expression and direct migration of pulmonary CD8+/CXCR3+ T cells in the lungs of patients with HIV infection and T-cell alveolitis. AB - The recruitment of cytotoxic T lymphocytes (CTL) is considered to be the major tool for the clearance of HIV from the lower respiratory tract. In this study we evaluated the pathophysiologic role of two lymphotactic CXC chemokines (IP-10 and Mig) in the lung of HIV-infected patients. These chemokines stimulate the directional migration of activated T cells and interact with a specific receptor (CXC receptor 3, CXCR3). Lymphocytes recovered from the bronchoalveolar lavage (BAL) of HIV-infected patients with high intensity T-cell alveolitis were CD8+ T cells expressing high levels of CXCR3 and IFN-gamma, a phenotype that is characteristic of Tc1 cells. Pulmonary T cells expressing CXCR3 exhibited a high migratory capability in response to IP-10 and Mig. Alveolar macrophages recovered from patients with T-cell alveolitis bore the IFN-gamma-inducible proteins IP-10 and Mig. A positive correlation was demonstrated between IP-10, Mig, and IL-15 expression by alveolar macrophages. Interestingly, macrophages isolated from the lung of HIV-infected patients with T-cell alveolitis secreted definite levels of CXCR3 ligands capable of inducing T-cell chemotaxis. Taken together, our data suggest that chemotactic ligands that bind CXCR3 contribute significantly to the accumulation of HIV-specific CTL in the lung. PMID- 11029364 TI - Sleep-disordered respiration in phenotypically normotensive, genetically hypertensive rats. AB - Increased prevalence of sleep-related breathing disorders has been reported in patients with essential hypertension and we have described disordered breathing in spontaneously hypertensive rats, an animal model of genetic hypertension. The mechanisms coupling hypertension to respiratory dysfunction during sleep remain, however, largely unknown. To determine if sleep-related respiratory disorder reflects cardiovascular derangement or, alternatively, represents an independent phenotype in hypertensive rats, we polygraphically recorded groups (n = 10) of genetically hypertensive, genetically normotensive, and phenotypically normotensive rats carrying a genetic background for hypertension. Apnea index was elevated more than 15-fold during NREM sleep in both animal groups carrying hypertension-related genes (p < 0.0001 for each) versus normotensive Wistar Kyoto rats. During REM sleep, a genetic background for hypertension was associated with an increased apnea index of at least 500% versus normotensive Wistar Kyoto rats (p < 0.0001 for each comparison). Still, overall mean respiratory rate, minute ventilation, and sleep architecture were equivalent among all animal groups. As expected, blood pressure and heart period were similar in both normotensive groups but elevated in the hypertensive animals. Persistent sleep-related breathing disorder despite effective cardiovascular normalization in the phenotypically normotensive but genetically hypertensive rats suggests that disordered breathing represents a genetically determined phenotype in these animals that is not secondary to the cardiovascular derangements. The model system described here may provide a powerful tool for investigation of the determinants of sleep-related breathing disorder. PMID- 11029365 TI - Reduced alpha- and beta(2)-adrenergic vascular response in patients with obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) has been associated with increased sympathetic activity. This study tested the hypothesis that the alpha- and beta(2)-receptor mediated vascular response is altered in patients with OSA. Forearm vascular resistance was evaluated by venous occlusion plethysmography in 10 normotensive OSA patients and 10 normotensive controls (apnea/hypopnea index [mean +/- SD] 29.4 +/- 2.3 and 1.6 +/- 0.3 per hour, respectively) roughly matched for body mass index (BMI) and age. Forearm vascular resistance was measured after intraarterial infusion of norepinephrine (NE) (7.4, 31, 120, 472 and 1421 pmol/100 ml forearm volume [FAV]/min), before and after phentolamine infusion (2 microgram/100 ml FAV/min), and isoproterenol (ISO) (1, 2, 6, and 15 ng/100 ml FAV/min). NE-induced vasoconstriction was significantly attenuated in OSA patients compared with controls (65.0 +/- 36.6% versus 129.4 +/- 81.8%, p = 0.049). The reduction of vascular resistance after phentolamine was similar in patients and control subjects (-50.8 +/- 16.7% versus -43.4 +/- 20.0%, p = 0.38). During ongoing phentolamine infusion NE increased resistance to a similar extent in both groups (0.5 +/- 4.9% versus -0.9 +/- 10.1%, p = 0.96). Vasodilation following ISO was significantly attenuated in OSA patients compared with control subjects (-53.3 +/- 9.0% versus -64.7 +/- 10.3%, p = 0.049). Moreover, the vascular response to NE in OSA patients was negatively correlated with plasma NE concentration (r = -0.76, p < 0.05). The reduced vascular response to alpha- and beta-receptor stimulation suggests a functional downregulation of vascular sympathoadrenergic receptors in patients with sleep apnea. PMID- 11029366 TI - Enhanced levels of whole-body protein turnover in patients with chronic obstructive pulmonary disease. AB - A substantial number of patients with chronic obstructive pulmonary disease (COPD) are characterized by fat-free mass wasting and altered muscle and plasma amino acid levels, suggesting changes in protein metabolism. In the present study, we examined whether whole-body protein breakdown (PB) and synthesis (PS) differ between 14 stable patients with COPD and 8 healthy controls. Whole-body PB, PS, and net PB (= PB-PS) were measured by the combined infusion of the stable isotopes L-[ring-(2)H(5)]phenylalanine (Phe) and L-[ring-(2)H(2)]tyrosine. Because there is evidence for specific disturbances in leucine (Leu) metabolism, the PB values were compared with those obtained when infusing L-[1-(13)C]Leu tracer. In arterialized-venous plasma and in vastus lateralis muscle, the isotope enrichment values and amino acid concentrations were measured. Whole-body PS and PB, assessed by Phe and Tyr tracer, were higher in the COPD group than in the control group (p < 0.05), indicating an elevated protein turnover. Net PB was increased in both groups, indicating a comparable degree of protein catabolism in the postabsorptive state. In contrast, whole-body PB determined by Leu tracer was not different between the groups. As a consequence, the ratio of Leu to Phe breakdown was reduced in the COPD group (p < 0.001). Moreover, in the COPD group a higher muscle-to-plasma gradient was found for Leu (p < 0.001) but not for Phe. The present study reveals elevated levels for protein turnover in patients with COPD, and indicates that infusion of the Leu tracer gives a reflection of Leu metabolism but not of whole-body protein metabolism in these patients. PMID- 11029367 TI - Imbalance between platelet vascular endothelial growth factor and platelet derived growth factor in pulmonary hypertension. Effect of prostacyclin therapy. AB - Focal vascular injury and impaired endothelial function are features of pulmonary hypertension (PH) that lead to enhanced platelet endothelial cell interactions. Vascular endothelial growth factor (VEGF) is contained in platelets and released at sites of vascular injury to promote endothelial repair and wound healing in combination with platelet-derived nonspecific mitogens such as platelet-derived growth factor (PDGF). The overall balance between platelet VEGF and PDGF was investigated in 21 patients with primary PH, 8 with secondary PH, and 27 with chronic hypoxemic lung disease (CHLD), as well as in 29 control subjects. Platelet VEGF content was increased in patients with primary and secondary PH as compared with control subjects (518 +/- 89, 675 +/- 156, and 166 +/- 29 fg/10(5) platelets, respectively; p < 0.01), whereas platelet PDGF content was similar in the three groups (31 +/- 2, 36 +/- 4, and 33 +/- 3 pg/10(5) platelets, respectively; NS). Patients treated with a continuous prostacyclin infusion had a higher platelet VEGF but a similar platelet PDGF content as compared with untreated patients. Moderate increases in platelet VEGF and PDGF contents were observed in the CHLD patients. We conclude that patients with primary or secondary PH have an increase in platelet VEGF content, but not in platelet PDGF content, and that their platelet VEGF content increases further in response to prostacyclin infusion. We suggest that imbalance between platelet VEGF and PDGF is beneficial to patients with PH. PMID- 11029368 TI - The effect of inhaled budesonide on symptoms, lung function, and cold air and methacholine responsiveness in 2- to 5-year-old asthmatic children. AB - We hypothesized that measurement of lung function (LF) and bronchial hyperresponsiveness (BHR) could serve as supplemental tools in evaluating the efficacy of treatment with inhaled corticosteroids in asthmatic children aged 2 to 5 yr. We studied 38 children (mean age: 53 mo; range: 35 to 71 mo) with moderately severe asthma in a single-center, randomized, double-blind, parallel group, placebo-controlled study involving 8 wk of treatment. Budesonide (BUD) 400 microgram twice daily was administered via a pressurized metered-dose inhaler and metal spacer device. Symptom scores (SSc) and use of short-acting beta(2)-agonist were monitored with diary cards. LF in awake children was measured as the specific airway resistance (sRaw), using whole-body plethysmography; as resistance by the interrupter technique (Rint); and as resistance and reactance at 5 Hz (Rrs5, Xrs5) by the impulse oscillation technique. Cold air challenge (CACh) and methacholine challenge (MCh) were used to assess BHR. Children in the BUD group experienced significantly fewer night- and daytime symptoms (p < 0.05) and more symptom-free days (p < 0.05), but not nights (p = 0.07), than children in the placebo group. Daytime (p < 0.05) but not nighttime (p = 0.09) use of rescue medication and asthma exacerbation rates (3.7 versus 9.3 exacerbations/yr) (p = 0.006) were both in favor of BUD. LF measured with the Rint technique, Rrs5, and Xrs5 were significantly improved by BUD. BHR as measured by CACh improved significantly with BUD, whereas no improvement was found on MCh. In conclusion, inhaled BUD at a total dose of 800 microgram daily significantly improved SSc, asthma exacerbation rates, lung function, and BHR as assessed by CACh in asthmatic children aged 2 to 5 yr. PMID- 11029369 TI - Usefulness of sniff nasal pressure in patients with neuromuscular or skeletal disorders. AB - Inspiratory muscle strength is an important variable in patients with neuromuscular or skeletal disorders. It is usually assessed by measuring maximal inspiratory pressure (PI(max)), but this test may prove difficult for some patients, and low values may originate from incomplete effort or air leaks. We assessed the usefulness of the novel sniff nasal pressure (Pn(sn)) test in 126 patients with a neuromuscular or a skeletal disorder, aged 5 to 49 yr. Pn(sn) was measured in an occluded nostril during maximal sniffs performed through the contralateral nostril. All patients performed the Pn(sn) maneuver easily, whereas 10 young and weak patients with neuromuscular disorders could not perform the PI(max) maneuver. Data were analyzed for the 116 patients who could perform both tests (92 patients with neuromuscular and 24 with skeletal disorders). When expressed as percents of the predicted values, Pn(sn) was similar to PI(max) in patients with neuromuscular disorders (54 +/- 25% predicted [mean +/- SD] versus 52 +/- 24% predicted), and was higher than PI(max) in patients with skeletal disorders (70 +/- 25% predicted versus 61 +/- 27% predicted, p < 0.05). Pn(sn) appeared to be the main determinant of VC in patients with neuromuscular disorders, whereas the Cobb angle and PI(max) were the main determinants of VC in patients with skeletal disorders. We conclude that inspiratory muscle strength can be easily assessed with Pn(sn) in children and adults with various neuromuscular and skeletal disorders. This new muscular parameter appears particularly useful in neuromuscular disorders, in which it represents a major determinant of VC. PMID- 11029370 TI - Does snoring predict sleepiness independently of apnea and hypopnea frequency? AB - Obstructive apneas and hypopneas during sleep are a well recognized cause of excessive daytime sleepiness. Snoring is also associated with excess sleepiness, although it is not known whether this reflects an independent effect of snoring or whether snoring is simply a marker for obstructive sleep apnea. To further explore the relation of snoring to sleepiness, we conducted a cross-sectional cohort study of community-dwelling adults participating in the Sleep Heart Health Study. The study sample comprises 2,737 men and 3,040 women with a mean age of 64 (SD 11) yr. Sleepiness was quantified using the Epworth Sleepiness Scale (ESS). Snoring history was obtained via a self-completion questionnaire. The respiratory disturbance index (RDI), defined as the number of apneas plus hypopneas per hour of sleep, was measured during in-home polysomnography. The ESS score increased progressively with increasing RDI, from a mean of 7.1 (4.2) in subjects with RDI < 1.5 to 8.8 (4.8) in subjects with RDI >/= 15 (p < 0.001). A progressive increase in ESS score was also seen across five categories of snoring frequency, from 6.4 (4.2) in current nonsnorers to 9.3 (4.8) in subjects who snored six to seven nights per week (p < 0.001). The prevalence of excessive daytime sleepiness, defined as an ESS score >/= 11, increased from 15% in never-snorers to 39% in those who snored six to seven nights per week. The relation of snoring to sleepiness was seen at all levels of RDI, with no significant change in the relation of snoring to ESS score after adjustment for RDI in multivariate models. The effects of snoring and RDI on sleepiness were little affected by adjustment for age, sex, race, body mass index, or questionnaire evidence of insufficient sleep time or nocturnal leg jerks or cramps. We conclude that both snoring and RDI are independently associated with excess sleepiness in community-dwelling, middle-aged and older adults. PMID- 11029371 TI - Airway wall thickness in asthma assessed by computed tomography. Relation to clinical indices. AB - Postmortem studies have shown that airway wall thickening is present in asthmatic patients and may play a pathophysiologic role. We investigated the presence and characteristics of airway wall thickening in patients with asthma, using helical computed tomography. Eighty-one asthmatic patients and 28 healthy control subjects were studied cross-sectionally. Airway wall thickness was assessed by a validated method on the basis of wall area (WA), WA corrected by body surface area (WA/BSA), and WA%, defined as (WA/total area) x 100 at the apical bronchus of the right upper lobe. Airway luminal area (Ai) and Ai/BSA were also examined. Asthma duration and severity, pulmonary function, and serum eosinophil cationic protein levels were evaluated. Intraobserver and interobserver reproducibility of WA, WA%, and Ai measurements were good. As compared with control, WA, WA/BSA, and WA% were significantly increased in patients with mild (n = 13), moderate (39), and severe persistent (22) asthma but not in patients with intermittent asthma (7). Comparison of the four asthmatic subgroups demonstrated thicker airways in more severe disease, but no difference in Ai or Ai/BSA. When all asthmatic patients were analyzed together, WA and WA/BSA correlated with the duration, although weakly, and severity of asthma. WA and WA/BSA negatively correlated with FEV(1) (percentage of predicted), FEV(1)/FVC (%), and FEF(25-75%) (percentage of predicted), whereas WA% negatively correlated with only FEV(1). We conclude that airway wall thickening occurs in patients with asthma and is not limited to those with severe disease. The degree of airway wall thickening may relate to the duration and severity of disease and the degree of airflow obstruction. PMID- 11029372 TI - Commercial versus native surfactants. Surface activity, molecular components, and the effect of calcium. AB - Despite their broad clinical use, there is no standardized comparative study on the functional, biochemical, and morphologic differences of the various commercial surfactants in relation to native surfactant. We investigated these parameters in Alveofact, Curosurf, Exosurf, and Survanta, and compared them with native bovine (NBS) and porcine (NPS) surfactant. For Curosurf and Alveofact the concentrations necessary for minimal surface tensions < 5 mN/m were six to 12 times higher (1.5 and 3 mg/ml, respectively) than with NPS and NBS. Exosurf and Survanta only reached 22 and 8 mN/m, respectively. Increasing calcium to nonphysiologic concentrations artificially improved the function of Alveofact and Curosurf, but it had little effect on Exosurf and Survanta. Impaired surface activity of commercial versus native surfactants corresponded with their lack in surfactant protein SP-A and decreased SP-B/C. The higher surface activity of Curosurf compared with Alveofact corresponded with its higher concentration of dipalmitoylphosphatidylcholine (DPPC). Despite their enrichment in DPPC Survanta and Exosurf exhibited poor surface activity because of low or absent SP-B/C. Ultrastructurally, Curosurf and Alveofact consisted mainly of lamellar and vesicular structures, which were also present in NPS and NBS. Exosurf contained crystalline structures only, whereas the DPPC-enriched Survanta contained separate lamellar/vesicular and crystalline structures. We conclude that in vitro surface activity of commercial surfactants is impaired compared with native surfactants at physiologic calcium concentrations. In the presence of SP-B/C, surface activity corresponds to the concentration of DPPC. Our data underscore the importance of a standardized protocol at physiologic calcium concentrations for the in vitro assessment of commercial surfactants. PMID- 11029373 TI - Measurement of pleural pressure during therapeutic thoracentesis. AB - The aim of therapeutic thoracentesis (TT) is to aspirate as much pleural fluid as possible. Monitoring pleural pressure (PlP) during TT has been proposed to avoid the adverse effects due to an unintended sharp drop in PlP. The objectives of this study are to ascertain the diagnostic value of the PlP measurement, to find a predictive variable of the amount of fluid that can be removed, to obtain insight into the characteristics of the PlP curve and pleural elastance (PE) during TT, and to describe the complications of TT. Sixty-one unselected patients were studied. Only the four patients with suspected trapped lung had an initial PlP lower than -4 cm H(2)O and a PE higher than 33 cm H(2)O/L. There was a weak correlation (r = 0.52) between PE during the first 0.5 L aspirated and the total amount of fluid aspirated. Partial PE values were 10, 7.5, and 14 cm H(2)O/L at the early, intermediate, and late phases of TT. No complications were found except for nine pneumothoraces. In conclusion, the technique was clinically helpful because large amounts of pleural fluid could be aspirated with few and mild complications, and because it allows clinicians to support the preliminary diagnosis of trapped lung. None of the studied variables was found to predict the suitability of aspirating more than 1.5 L. Rather than being monotonically descendent, the PlP curve shows a three-part line with the deepest slopes at the first and last phases of the thoracentesis. PMID- 11029374 TI - Albumin-mediated regulation of cellular glutathione and nuclear factor kappa B activation. AB - Human serum albumin (HSA) is a cystine-rich serum protein taken up by many cells through receptor-mediated and fluid-phase endocytosis. We hypothesized that HSA may play a role in modulating cellular antioxidant redox signaling. Lung epithelial cells (A549), fibroblasts (HFL1), and blood lymphocytes had increased glutathione (GSH) levels after 8 h incubation with HSA. Similar GSH increases were observed with either plasma-derived or recombinant HSA. Serum depleted of HSA had no effect on cellular GSH. The GSH increase was also observed in normal murine lungs upon in vivo airway instillation of HSA. GSH enhancement was not related to the redox state of the free cysteine residue (Cys-34) on HSA, however, reduction of disulfide bonds in HSA inhibited the increase in cellular GSH. In addition, the albumin-mediated increase in GSH was inhibited by the vacuolar (H(+))-ATPase inhibitors, bafilomycin A(1) and concanamycin, as well as by the membrane pH-disrupting ionophore monensin, but not by 20 mM NH(4)Cl. The degree to which albumin increased GSH levels was sufficient to protect cells against H(2)O(2)-mediated cytotoxicity and to decrease TNF-alpha-mediated NF-kappaB activation. We conclude that albumin specifically modulates cellular GSH levels, an effect sufficient to protect cells against oxidant injury and regulate NF kappaB activation. PMID- 11029375 TI - Mechanisms of endotoxin-induced airway and pulmonary vascular hyperreactivity in mice. AB - Endotoxin is thought to contribute to pulmonary hyperresponsiveness in byssinosis, asthma, and the acute respiratory distress syndrome (ARDS). The aim of this study was to elucidate the mechanism of this phenomenon in the isolated, blood-free perfused mouse lung. Perfusion with lipopolysaccharide (LPS) had no effect on pulmonary resistance or pulmonary artery pressure, but induced airway hyperreactivity (AHR) to methacholine (MCh) and pulmonary vascular hyperreactivity (VHR) to platelet-activating factor (PAF). Blockade of the thromboxane/endoperoxide (TP) receptor with SQ29.548 completely protected against LPS-induced AHR and VHR. Blockade of cyclooxygenase-2 (COX-2) abolished LPS induced VHR but suppressed LPS-induced AHR only marginally. COX-2 messenger RNA was upregulated in LPS-treated lungs, and inhibition of transcription with actinomycin D or of protein biosynthesis with cycloheximide protected against LPS induced VHR but not AHR. Pretreatment with the radical scavenger N-acetylcysteine partly protected against LPS-induced AHR. In addition, perfusion of mouse lungs with the isoprostane 8-epiprostaglandin F(2alpha) (8-epi-PGF(2alpha)), which may be formed as a consequence of oxidative stress in the lung, elicited AHR, which was completely blocked by SQ29.548. Enzyme immunoassay did not detect either 8 epi-PGF(2alpha )or thromboxane B(2) in perfusate samples. Our findings show that LPS induces AHR and VHR in mouse lungs via activation of the TP receptor. Although induction of VHR depends on COX-2 activity, AHR is largely mediated by a non-COX-derived TP agonist, which might be a product of radical-induced lipid peroxidation. PMID- 11029376 TI - Induction of lung fibroblast apoptosis by soluble fibronectin peptides. AB - Despite the importance of fibroproliferative lung disorders, no safe and effective therapies exist for reducing the size of the fibroblast population in existing fibrotic lesions. Recent work suggests that therapies that promote fibroblast apoptosis during the repair phase following lung injury may facilitate lung repair by eliminating excess fibrotic tissue. We report here our finding that three soluble fibronectin peptides (RGD, CS-1, and FN-C/H-V) induce apoptosis in lung fibroblasts. Fibroblast susceptibility to these peptides was dose and time dependent, with a maximal effect observed at 96 h (87 +/- 16% [mean +/- SEM] apoptosis). The peptides were able to induce fibroblast apoptosis in fibrin gels, an in vitro model of early fibroproliferative lesions. Fibroblasts were difficult to kill. All three peptides were required for maximal apoptosis of anchored cells. Apoptosis occurred by disruption of adhesion (anoikis). Treatment of fibroblasts with peptides caused proteolysis of pp125FAK, a tyrosine kinase involved in integrin-mediated signaling related to cell survival. These data show that soluble fibronectin peptides trigger nontransformed fibroblast apoptosis in routine culture and in fibrin gels by a mechanism that includes disruption of an integrin-mediated survival signaling pathway. The use of small fibronectin peptides to promote fibroblast apoptosis warrants further study as possible antifibrotic therapy. PMID- 11029377 TI - Antisense oligonucleotides to NF-kappaB improve survival in bleomycin-induced pneumopathy of the mouse. AB - We examined the effect of antisense oligonucleotides to the p65 subunit of NF kappaB on the survival of bleomycin (BLM)-induced pneumonitis in C57BL/6 mice (female, 8 wk of age, 17 to 20 g body weight). Fifty-three percent and all control mice died within 6 to 9 d after intravenous administration of 150 and 300 mg/kg BLM alone, respectively. The intravenous administration of the antisense oligonucleotides (900 microgram/animal dissolved in 200 microliter saline, 6 h before and 5 d after BLM administration) significantly improved the survival rate to 100 and 40% in 150- and 300-mg/kg-treated animals, respectively. The antisense oligonucleotides also significantly improved the loss of body weight, the increase in lung hydroxyproline, and histologic changes by BLM, whereas the antisense oligonucleotides themselves did not produce any significant changes in the behavior or lung histology. Both peripheral blood monocytes and alveolar macrophages were confirmed to contain large amounts of intracellular antisense oligonucleotides after BLM injection by FITC-labeled fluorescent microscopy. Further, Western blotting confirmed the inhibition of NF-kappaB in macrophages by the antisense oligonucleotides. These findings suggest that the antisense oligonucleotides are incorporated into activated alveolar macrophages and ameleriorate the lung injury and pneumonitis/fibrosis, thereby improving the survival of BLM-induced pneumopathy in mice. PMID- 11029378 TI - Changes in extracellular matrix and tissue viscoelasticity in bleomycin-induced lung fibrosis. Temporal aspects. AB - Bleomycin-induced lung fibrosis results in changes in tissue mechanical properties due to alterations in the extracellular matrix (ECM). How oscillatory mechanics and changes in the matrix evolve over time has not been addressed. Sprague-Dawley rats were instilled with bleomycin sulfate (BM) (1.5 U) intratracheally; control animals (C) received saline. At 7, 14, and 28 d after BM, parenchymal strips (7 x 2 x 2 mm) were obtained and strips suspended in a Krebs-filled organ bath. One end of the strip was attached to a force (F) transducer and the other to a lever arm that effected sinusoidal length (L) oscillations. Strips were oscillated at varying amplitudes (1, 3, and 10% of resting L) and frequencies (f = 0.3, 1, 3, and 10 Hz) at an operating stress of 2 kPa. Resistance (R) and elastance (E) were estimated by fitting changes in F and L to the equation of motion. Hysteresivity (eta) was calculated as eta = (R/E) 2pif. Strips were then fixed for morphological study of collagen, elastic fibers, and the small proteoglycans (PGs), biglycan and fibromodulin (FM). R and E were significantly greater and eta significantly less in BM versus C strips (p < 0.001). The increase in R and E peaked at 14 d after BM; the decrement in eta was maximal at Day 7. Biglycan was increased in BM lung strips at all time points, FM and elastic fibers were increased at 14 and 28 d, and collagen was increased at 28 d only. Hence, changes in mechanics were maximal before collagen content had increased. In addition, we demonstrated a significant correlation between biglycan and all mechanical parameters. These data suggest that changes in PGs may be critical in determining changes in lung tissue viscoelastic behavior in this fibrosis model PMID- 11029379 TI - Distribution of obstructive intimal lesions and their cellular phenotypes in chronic pulmonary hypertension. A morphometric and immunohistochemical study. AB - We investigated the distribution of pulmonary arteriopathy in chronic pulmonary hypertension (PH) in a quantitative histopathologic study, using computer assisted image analysis. We also examined the histologic manifestations and cellular phenotypes of various obstructive intimal lesions in PH with an immunohistochemical method. A total of 53 lungs removed at autopsy or explantation were obtained for the study from 51 documented cases of moderate to severe PH (15 cases of primary pulmonary hypertension [PPH], eight cases of Eisenmenger's syndrome [EISEN], 22 cases of chronic major-vessel thromboembolic disease [CTED], and three cases of PH associated with other known causes), and two unused donor lungs served as normal controls. Intimal thickening in PPH was most prominent in small pulmonary arteries and arterioles less than 200 micrometer in diameter. Plexiform lesions in PPH were associated with significantly smaller arteries than in EISEN. Arteries larger than 400 micrometer showed a significant intimal thickening only in CTED. Obstructive intimal lesions in PH comprised a morphologic spectrum with frequent intermediate forms between plexiform and thrombotic lesions. Most cells within various intimal lesions showed an immunoprofile of myofibroblasts that were positive for vimentin and alpha-smooth muscle actin, but negative for desmin and endothelial markers including Factor VIII, clonal designator (CD)31, and CD34. Endothelial markers were positive only in the single layer of cells lining slitlike lumens, when the latter were present. In conclusion, major types of PH had characteristic distribution patterns of obstructive intimal lesions, showing mainly a myofibroblastic phenotype and variable endothelial/vascular differentiation. PMID- 11029381 TI - Purpose of the conference PMID- 11029380 TI - Linkage analysis of chromosome 12 markers in Italian families with atopic asthmatic children. AB - We investigated 116 Italian atopic families (560 individuals) for linkage with 13 DNA markers on chromosome 12. All the subjects were phenotyped for asthma, total serum IgE, bronchial hyperresponsiveness, skin-prick positivity to common aeroallergens, and atopy. A relative location map of the markers was prepared from Centre d'Etude du Polymorphisme Humain families. Affected sib pair multipoint linkage methods were used to perform the statistical analyses. We report suggestive linkage for asthma with markers on chromosome 12. The region of interest centers around marker D12S390 (maximum logarithm of odds [mlod] = 2.81; p = 0.003). These results provide additional support that asthma susceptibility factors are located on chromosome 12q. PMID- 11029382 TI - Chairman's summary. Antigen processing, presentation, and immunomodulation. PMID- 11029383 TI - Innate immune system damage in human immunodeficiency virus type 1 infection. Implications for acquired immunity and vaccine design. AB - HIV infection affects the innate as well as the acquired immune systems. Critically, it changes the function of macrophages, which link the innate and acquired responses through their ability to present antigen to CD4(+) T lymphocytes. Patients with HIV infection have a reduced capacity to deal with subsequent pathogen exposure and many suffer from chronic pulmonary infections. We have produced complex synthetic peptides that mimic the function of viral gp120 and may represent prototypes of molecules that can prevent or ameliorate HIV-induced damage to the immune system. PMID- 11029384 TI - Antigen-presenting cells. Experimental and clinical studies of dendritic cells. AB - We present a summary of our experimental and clinical studies of dendritic cells (DCs) in five different areas. First, human DCs undergo chemotaxis and transendothelial migration in response to distinct chemokines, depending on their maturation state; inducible chemokines may recruit immature DCs to sites of inflammation and infection, whereas constitutive chemokines may regulate subsequent migration to lymphoid tissues. Second, M-tropic but not T-tropic strains of HIV-1 induce and modulate DC chemotaxis in culture; conceivably this may facilitate viral infection and contribute to pathogenesis in vivo. Third, malaria-infected erythrocytes modulate the maturation and inhibit the function of DCs; adhesion of infected erythrocytes to DCs may impart a selective advantage to the parasite by modulating host immunity. Fourth, a phase I clinical trial of DC based immunotherapy of cancer has been initiated in which autologous DCs are administered directly into metastatic skin lesions; immune responses to a recall antigen, with which the cells were pulsed, were enhanced in several patients. Finally we describe the growth and characterization of a stably immature, maturation-resistant stage of DCs generated from mouse bone marrow that can induce transplantation tolerance in vivo, and outline studies of their capacity to induce responses of transgenic T cells in vivo. PMID- 11029385 TI - Antigen presentation in the lung. AB - Studies from our laboratory and elsewhere have implicated populations of dendritic cells in lung and airway tissues as key regulators of both qualitative and quantitative aspects of T cell responses to local antigenic challenge. Under steady state conditions, they are specialized for uptake of antigen, and require additional maturation signals for full expression of their T cell-stimulating activity. Their functional phenotype appears to be controlled via a complex series of interactions with both bone marrow-derived, mesenchymal, and possibly neuroendocrine cells; failure(s) in one or more of these regulatory interactions may be important etiologic and/or pathogenic factors in a variety of respiratory immunoinflammatory disease. PMID- 11029386 TI - Regulation of helper T cell differentiation and recruitment in airway inflammation. AB - CD4(+) helper T type 1 (Th1) and Th2 cells are critical mediators of inflammatory diseases. Although T cells represent only a fraction of the leukocytes that are found in the lung during inflammation, they play a critical role in coordinating the immune response to infectious agents and allergens. T cells have the ability to rapidly expand in response to specific stimuli and to differentiate into effector cells that, through the production of soluble factors such as cytokines and chemokines, communicate with other cells to initiate a cascade of inflammatory events. The objective of this review is to outline the cellular and molecular mechanisms involved in the generation and recruitment of Th1 and Th2 cells in the lung. Defining these mechanisms should lead to improved immunopharmacological strategies for prophylaxis and therapy. PMID- 11029387 TI - gammadelta cells and the regulation of mucosal immune responses. AB - We are only now uncovering the potentially important contributions made to immune responses by gammadelta cells. These contributions are likely to be particularly important at mucosal sites, where gammadelta cells are disproportionately enriched. Indeed, gammadelta cells have proven biological activity in the lung. In addition, gammadelta cells are also enriched in young rather than adult animals. Studies of mutant mice have demonstrated that alphabeta T cells are seemingly essential for high-affinity, cognate immunological memory, whereas gammadelta cells contribute to the early stages of an immune response and to the regulation of alphabeta T cell- and B cell-mediated immunity. To explore further the role of gammadelta cells in immune responses, we have investigated whether their contribution is greater during the early period of life, when the cells are more abundant. In a natural system of coccidial infection of gut epithelial cells, we find that alphabeta T cell responses are less essential for immunoprotection during primary challenge of young mice than is true for adult animals. This "ineffectiveness" creates a "window of importance" for the immunoprotective capacity of gammadelta cells, which seem thereby to be more crucial in young compared with older animals. The relative ineffectiveness of alphabeta T cells in young mice may be attributable to a bias toward Th2 activity. We therefore hypothesize that gammadelta cell activity, elicited by infection early in life, may compensate for defects in Th1 activity and may actually accelerate the bias in alphabeta T cells away from Th2. This has obvious implications for susceptibility to Th2-type allergic responses. PMID- 11029388 TI - Costimulation of T cells. AB - Costimulation is critical to T cell activation. On the antigen-presenting cell the key players are found in the extended family of B7 genes comprising cd80, cd86, B7h/B7RP-1 and B7-H1. cd80 and cd86 encode proteins that bind to CD28 and CTLA4 on T cells. Blocking this pathway with the potent CTLA4-Ig fusion protein shows encouraging potential as a therapeutic agent. While cd80 and cd86 pathways act mainly on naive T cells, B7h/B7RP-1 and B7-H1 seem to exert their effects on antigen-experienced lymphocytes. PMID- 11029389 TI - Immunological tolerance to inhaled antigen. AB - Regulatory mechanisms exist in the immune system to limit the induction of pathogenic responses to antigens encountered within the respiratory tract. The development of allergic disease is thought to arise as a result of the breakdown in these regulatory processes. In this review we examine the nature of immune responses generated to inhaled protein antigens and the mechanisms used to establish tolerance to inhaled antigens. PMID- 11029390 TI - Oral tolerance. AB - The intestinal immune system discriminates between potentially harmful and harmless foreign proteins. The basis for this differential response may be related to the conditions of antigen presentation by antigen-presenting cells, as determined by their phenotype or activation state. How these conditions affect specific immunologic unresponsiveness to later challenge with an antigen is not known. Two possible mechanisms are the induction of anergy or deletion of responsive cells and the activation of regulatory cells or mediators, and the mechanism may very depending on the tolerizing regimen used. Should regulatory cells be involved, they are speculated to induce tolerance through their production of inhibitory cytokines, such as IL-4, IL-10, and TGF-beta. Studies using specific antibodies and selective genetic knockout (KO) strains of mice, however, have provided conflicting data. A final intriguing possibility is that tolerance results from cognate interactions between T cells and APCs, so that tolerant T cells or APCs prime T cells they contact to deliver a tolerogenic signal to the next T cell they encounter, possibly through a function dependent on interactions between Notch family receptors and their ligands. As with many questions in mucosal immunology, definition of the mechanisms of oral tolerance (OT) has proved difficult to address experimentally, but promising approaches include study of the distribution of fed antigen, of targeted genetic KOs, and of transgenic strains. PMID- 11029391 TI - Biotherapeutic targets for the treatment of allergic airway disease. AB - T cells are critical mediators of inflammation and as such, their migration to inflammatory sites is a tightly controlled process involving a complex series of molecules expressed by a variety of cell types. As our appreciation of the mechanisms governing T cell surveillance, activation, differentiation, and subsequent homing to sites of inflammation has advanced, the opportunity to develop novel therapeutic agents that modulate the immune system has increased. Importantly, the possibility of specifically targetting subpopulations of effector cells raises the exciting potential for the development of novel agents that selectively modify the immune response to allergens, without resulting in generalized immune suppression. PMID- 11029392 TI - Control of experimental inflammatory bowel disease by regulatory T cells. AB - A helper T cell type 1-mediated colitis driven by enteric bacteria develops in severe combined immunodeficient mice after transfer of CD45RB(high)CD4(+) T cells. Development of disease can be prevented by cotransfer of the reciprocal CD45RB(low) subset. Analysis of the mechanism of immune suppression transferred by CD45RB(low)CD4(+) cells revealed essential roles for both IL-10 and TGF-beta. These data indicate that a functionally specialized population of regulatory T (Treg) cells exists in normal mice and that these can prevent the development of pathogenic responses toward commensal bacteria. The role of Treg cells in the control of the immune response is discussed. PMID- 11029393 TI - Antigen presentation and DNA vaccines. AB - There is reasonable evidence that both cross-priming and direct transfection of antigen-presenting cells (APCs) play a role in induction of immune responses by DNA vaccines. It is not known which mode is more important for priming cytotoxic T cell responses, but both are sufficient and neither alone is necessary. Hence, a rational strategy for increasing DNA vaccine potency would be to facilitate both pathways. With regard to cross-priming, a better understanding of the nature of the antigen transferred and the molecules/cells involved may suggest ways to design DNA vaccines to enhance this pathway. With respect to transfection of APCs, certain DNA formulations or delivery systems may be able to target APCs for increased DNA uptake. Other considerations include recruitment of APCs to the site of DNA injection and manipulation of these cells to ensure the proper activation state for priming immune responses. The burgeoning scientific literature in these areas indicates that much effort is currently being directed toward these goals. PMID- 11029394 TI - Gene delivery to the corneal endothelium. AB - Gene transfer to the corneal endothelium has potential for modulating rejection of corneal grafts. It can also serve as a convenient and useful model for gene therapy of other organs. In this article we review the work carried out in our laboratory using both viral and nonviral vectors to obtain gene expression in the cornea. PMID- 11029395 TI - Cell logic for dual coupling of a single class of receptors to G(s) and G(i) proteins. PMID- 11029396 TI - GATA-6: the proliferation stops here: cell proliferation in glomerular mesangial and vascular smooth muscle cells. PMID- 11029397 TI - Increased Na(+)-Ca(2+) exchanger in the failing heart. PMID- 11029398 TI - Selective downregulation of VEGF-A(165), VEGF-R(1), and decreased capillary density in patients with dilative but not ischemic cardiomyopathy. AB - Cardiomyopathy (CM) comprises a heterogeneous group of diseases, including ischemic (ICM) and dilative (DCM) forms. The pathogenesis of primary DCM is not clearly understood. Recent studies in mice show that vascular endothelial growth factor (VEGF) is involved in ICM. Whether VEGF plays a role in human CM is unknown. We examined the mRNA and protein expression of VEGF and its receptors in hearts of patients with end-stage DCM and ICM and in healthy individuals using real-time polymerase chain reaction and Western blotting. Number of capillaries, area of myocytes, and collagen were calculated in cardiac biopsies using transmission electron microscopy. In DCM, except for VEGF-C, mRNA transcript levels of VEGF-A(165), VEGF-A(189), and VEGF-B and the protein level of VEGF-A and VEGF-R(1) were downregulated compared with controls (P:<0.05). However, in ICM, mRNA transcript levels of VEGF isoforms and protein levels of VEGF-C were upregulated. The vascular density was decreased in DCM but increased in ICM compared with controls (P:<0. 05). Muscular hypertrophy was not different for ICM and DCM, although DCM had more collagen (P:<0.05). Blunted VEGF-A and VEGF-R(1) protein expression and downregulated mRNA of the predominant isoform of VEGF-A, VEGF-A(165), to our knowledge shown here for the first time, provide evidence that the VEGF-A defect in DCM is located upstream. Whether downregulation of certain VEGF isoforms in DCM is a cause or consequence of this disorder remains unclear, although upregulated VEGF levels in ICM are most likely the result of ischemia. PMID- 11029399 TI - Microvascular integrity and the time course of myocardial sodium accumulation after acute infarction. AB - Loss of membrane permeability caused by ischemia leads to cellular sodium accumulation and myocardial edema. This phenomenon has important implications to left ventricular structure and function in the first hours after myocardial infarction. We hypothesized that during this period of time, after prolonged coronary occlusion and complete reflow, the rate of myocardial sodium accumulation is governed by microvascular integrity. We used 3-dimensional (23)Na MRI to monitor myocardial sodium content changes over time in an in vivo closed chest canine model (n=13) of myocardial infarction and reperfusion. Infarcts with microvascular obstruction (MO) defined by both radioactive microspheres and contrast-enhanced (1)H MRI showed a slower rate of sodium accumulation as well as lower blood flow at 20 minutes and 6 hours after reperfusion. Conversely, the absence of MO was associated with faster rates of sodium accumulation and greater blood flow restoration. In addition, infarct size by (23)Na MRI correlated best with infarct size by triphenyltetrazolium chloride and contrast-enhanced (1)H MRI at 9 hours after reperfusion. We conclude that in reperfused myocardial infarction, sodium accumulation is dependent on microvascular integrity and is slower in regions of MO compared with those with patent microvasculature. Finally, (23)Na MRI can be a useful tool for monitoring in vivo myocardial sodium content in acute myocardial infarction. PMID- 11029400 TI - Dephosphorylation and intracellular redistribution of ventricular connexin43 during electrical uncoupling induced by ischemia. AB - Electrical uncoupling at gap junctions during acute myocardial ischemia contributes to conduction abnormalities and reentrant arrhythmias. Increased levels of intracellular Ca(2+) and H(+) and accumulation of amphipathic lipid metabolites during ischemia promote uncoupling, but other mechanisms may play a role. We tested the hypothesis that uncoupling induced by acute ischemia is associated with changes in phosphorylation of the major cardiac gap junction protein, connexin43 (Cx43). Adult rat hearts perfused on a Langendorff apparatus were subjected to ischemia or ischemia/reperfusion. Changes in coupling were monitored by measuring whole-tissue resistance. Changes in the amount and distribution of phosphorylated and nonphosphorylated isoforms of Cx43 were measured by immunoblotting and confocal immunofluorescence microscopy using isoform-specific antibodies. In control hearts, virtually all Cx43 identified immunohistochemically at apparent intercellular junctions was phosphorylated. During ischemia, however, Cx43 underwent progressive dephosphorylation with a time course similar to that of electrical uncoupling. The total amount of Cx43 did not change, but progressive reduction in total Cx43 immunofluorescent signal and concomitant accumulation of nonphosphorylated Cx43 signal occurred at sites of intercellular junctions. Functional recovery during reperfusion was associated with increased levels of phosphorylated Cx43. These observations suggest that uncoupling induced by ischemia is associated with dephosphorylation of Cx43, accumulation of nonphosphorylated Cx43 within gap junctions, and translocation of Cx43 from gap junctions into intracellular pools. PMID- 11029401 TI - Myocardial mechanics and collagen structure in the osteogenesis imperfecta murine (oim). AB - Because the amount and structure of type I collagen are thought to affect the mechanics of ventricular myocardium, we investigated myocardial collagen structure and passive mechanical function in the osteogenesis imperfecta murine (oim) model of pro-alpha2(I) collagen deficiency, previously shown to have less collagen and impaired biomechanics in tendon and bone. Compared with wild-type littermates, homozygous oim hearts exhibited 35% lower collagen area fraction (P:<0.05), 38% lower collagen fiber number density (P:<0.05), and 42% smaller collagen fiber diameter (P:<0.05). Compared with wild-type, oim left ventricular (LV) collagen concentration was 45% lower (P:<0.0001) and nonreducible pyridinoline cross-link concentration was 22% higher (P:<0.03). Mean LV volume during passive inflation from 0 to 30 mm Hg in isolated hearts was 1.4-fold larger for oim than wild-type (P:=NS). Uniaxial stress-strain relations in resting right ventricular papillary muscles exhibited 60% greater strains (P:<0.01), 90% higher compliance (P:=0.05), and 64% higher nonlinearity (P:<0.05) in oim. Mean opening angle, after relief of residual stresses in resting LV myocardium, was 121+/-9 degrees in oim compared with 45+/-4 degrees in wild-type (P:<0.0001). Mean myofiber angle in oim was 23+/-8 degrees greater than wild-type (P:<0.02). Decreased myocardial collagen diameter and amount in oim is associated with significantly decreased fiber and chamber stiffness despite modestly increased collagen cross-linking. Altered myofiber angles and residual stress may be beneficial adaptations to these mechanical alterations to maintain uniformity of transmural fiber strain. In addition to supporting and organizing myocytes, myocardial collagen contributes directly to ventricular stiffness at high and low loads and can influence stress-free state and myofiber architecture. PMID- 11029402 TI - Vascular matrix metalloproteinase-2-dependent cleavage of calcitonin gene-related peptide promotes vasoconstriction. AB - Matrix metalloproteinase (MMP)-2 has been historically associated with the process of vascular remodeling through the cleavage of extracellular matrix proteins. However, we recently found that MMP-2 also cleaves the endothelium derived peptide big endothelin-1, ET-1[1-38] and yields the novel vasoconstrictor ET-1[1-32]. We therefore investigated the effects of MMP-2 inhibitors as potential vasodilators. MMP inhibition with ortho-phenanthroline (0.3 to 30 micromol/L) induced vasorelaxation of isolated rat mesenteric arteries (maximum of relaxation=74.5+/-27.6% at 30 micromol/L). However, phosphoramidon (0.3 to 30 micromol/L), which inhibits some metalloenzymes, but not MMP-2, did not dilate the arteries. Selective inhibition of endogenous MMP-2 with the novel tissue permeable cyclic peptide CTTHWGFTLC (CTT, 10 micromol/L) also caused vasorelaxation (by 85+/-6%), whereas STTHWGFTLS (10 micromol/L), an inactive CTT analogue, did not dilate the arteries. Interestingly, the vasorelaxation that results from MMP-2 inhibition was endothelium-independent. Thus, we examined whether MMP-2 acted on peptides derived from the smooth muscle or the perivascular nerves. Recombinant human MMP-2 cleaved calcitonin gene-related peptide (CGRP) specifically at the Gly(14)-Leu(15) peptide bond and reduced the vasodilatory potency of CGRP by 20-fold. Inhibition of MMP-2 increased the amount of intact CGRP in arteries and enhanced vasorelaxation induced by anandamide, which stimulates CGRP release. Vasorelaxation in response to MMP-2 inhibition was abolished by CGRP[8-37], a selective CGRP receptor antagonist, and by capsaicin, which depletes arterial perivascular nerves of CGRP. We conclude that vascular MMP-2 cleaves endogenous CGRP and promotes vasoconstriction. These data suggest a novel mechanism of regulating the vasoactive and, possibly, the neurohormonal actions of CGRP and establish MMP-2 as a modulator of vascular function. PMID- 11029403 TI - Membrane estrogen receptor engagement activates endothelial nitric oxide synthase via the PI3-kinase-Akt pathway in human endothelial cells. AB - 17beta-Estradiol (E(2)) is a rapid activator of endothelial nitric oxide synthase (eNOS). The product of this activation event, NO, is a fundamental determinant of cardiovascular homeostasis. We previously demonstrated that E(2)-stimulated endothelial NO release can occur without an increase in cytosolic Ca(2+). Here we demonstrate for the first time, to our knowledge, that E(2) rapidly induces phosphorylation and activation of eNOS through the phosphatidylinositol 3 (PI3) kinase-Akt pathway. E(2) treatment (10 ng/mL) of the human endothelial cell line, EA.hy926, resulted in increased NO production, which was abrogated by the PI3 kinase inhibitor, LY294002, and the estrogen receptor antagonist ICI 182, 780. E(2) stimulated rapid Akt phosphorylation on serine 473. As has been shown for vascular endothelial growth factor, eNOS is an E(2)-activated Akt substrate, demonstrated by rapid eNOS phosphorylation on serine 1177, a critical residue for eNOS activation and enhanced sensitivity to resting cellular Ca(2+) levels. Adenoviral-mediated EA.hy926 transduction confirmed functional involvement of Akt, because a kinase-deficient, dominant-negative Akt abolished E(2)-stimulated NO release. The membrane-impermeant E(2)BSA conjugate, shown to bind endothelial cell membrane sites, also induced rapid Akt and consequent eNOS phosphorylation. Thus, engagement of membrane estrogen receptors results in rapid endothelial NO release through a PI3-kinase-Akt-dependent pathway. This explains, in part, the reduced requirement for cytosolic Ca(2+) fluxes and describes an important pathway relevant to cardiovascular pathophysiology. PMID- 11029404 TI - Laminar shear stress upregulates integrin expression: role in endothelial cell adhesion and apoptosis. AB - Laminar shear stress exerts important effects on endothelial cell (EC) function and inhibits apoptosis of ECs induced by various stimuli. The mechanism by which hemodynamic forces, such as shear stress, are transduced into cellular signaling is still not known. Located at the cell surface, integrins, which are required for cell adhesion and cell survival, are potential mechanotransducers. Therefore, we investigated the effect of shear stress on integrin expression in ECs. Shear stress time-dependently increased the mRNA expression of the fibronectin receptor subunits alpha(5) and beta(1) with a maximum at 6 hours (283+/-41% and 215+/-27% of control, respectively). In addition, the protein levels of the fibronectin receptor subunits alpha(5) and beta(1) were enhanced with a maximum at 12 hours of shear stress exposure (343+/-53% and 212+/-38% of control, respectively). The shear stress-induced upregulation of integrins is independent of nitric oxide. Furthermore, we confirmed the enhanced functional activity of alpha(5)beta(1) integrin expression by FACS analysis. As a functional consequence, human umbilical vein ECs, which were preexposed to shear stress, revealed a significantly increased attachment (178+/-10% of static controls) and a more pronounced extracellular signal-regulated kinase 1 and 2 activation in response to cell attachment. Finally, we demonstrated that shear stress requires RGD sensitive integrins to mediate its antiapoptotic effect. Taken together, these results define a novel mechanism by which shear stress may exert its atheroprotective effects via upregulation of integrins to support EC adhesion and survival. PMID- 11029405 TI - Enhanced Ca(2+)-activated Na(+)-Ca(2+) exchange activity in canine pacing-induced heart failure. AB - Defective excitation-contraction coupling in heart failure is generally associated with both a reduction in sarcoplasmic reticulum (SR) Ca(2+) uptake and a greater dependence on transsarcolemmal Na(+)-Ca(2+) exchange (NCX) for Ca(2+) removal. Although a relative increase in NCX is expected when SR function is impaired, few and contradictory studies have addressed whether there is an absolute increase in NCX activity. The present study examines in detail NCX density and function in left ventricular midmyocardial myocytes isolated from normal or tachycardic pacing-induced failing canine hearts. No change of NCX current density was evident in myocytes from failing hearts when intracellular Ca(2+) ([Ca(2+)](i)) was buffered to 200 nmol/L. However, when [Ca(2+)](i) was minimally buffered with 50 micromol/L indo-1, Ca(2+) extrusion via NCX during caffeine application was doubled in failing versus normal cells. In other voltage clamp experiments in which SR uptake was blocked with thapsigargin, both reverse mode and forward-mode NCX currents and Ca(2+) transport were increased >2-fold in failing cells. These results suggest that, in addition to a relative increase in NCX function as a consequence of defective SR Ca(2+) uptake, there is an absolute increase in NCX function that depends on [Ca(2+)](i) in the failing heart. PMID- 11029406 TI - Cyclin A downregulation and p21(cip1) upregulation correlate with GATA-6-induced growth arrest in glomerular mesangial cells. AB - The GATA-6 transcription factor is reported to be expressed in vascular myocytes. Because glomerular mesangial cells (GMCs) and vascular myocytes have similar properties, we examined whether GATA-6 was expressed in cultured GMCs and whether overexpression of GATA-6 induced cell cycle arrest in GMCs, using a recombinant adenovirus that expresses GATA-6 (Ad GATA-6). GATA-6 expression in GMCs was downregulated when quiescent GMCs were stimulated by serum to reenter the cell cycle. [(3)H]thymidine uptake was inhibited in GMCs infected with Ad GATA-6 in a dose- and time-dependent manner. The expression of cyclin A protein was decreased and that of the cyclin-dependent kinase inhibitor p21(cip1) was increased in GMCs infected with Ad GATA-6. Although the expression of p21(cip1) transcripts did not change remarkably, p21(cip1) protein was stabilized in GMCs infected with Ad GATA 6, suggesting a post-transcriptional regulation of p21(cip1) expression. Northern blot analysis showed that expression of the cyclin A transcript was decreased in Ad GATA-6-infected cells, whereas this decrease of cyclin A was not observed in GMCs derived from p21(cip1) null mice. Our results demonstrate that GATA-6 is endogenously expressed in GMCs and that overexpression of GATA-6 can induce cell cycle arrest. Our results also show that GATA-6-induced cell cycle arrest is associated with inhibition of cyclin A expression and p21(cip1) upregulation. Finally, our results indicate that the GATA-6-induced suppression of cyclin A expression depends on the presence of p21(cip1). PMID- 11029407 TI - Beta(2)-adrenergic and several other G protein-coupled receptors in human atrial membranes activate both G(s) and G(i). AB - Cardiac G protein-coupled receptors that couple to Galpha(s) and stimulate cAMP formation (eg, beta-adrenergic, histamine, serotonin, and glucagon receptors) play a key role in cardiac inotropy. Recent studies in rodent cardiac myocytes and transfected cells have revealed that one of these receptors, the beta(2) adrenergic receptor (AR), also couples to the inhibitory G protein Galpha(i) (activation of which inhibits cAMP formation). If beta(2)ARs could be shown to couple to Galpha(i) in the human heart, it would have important ramifications, because levels of Galpha(i) increase with age and in failing human heart. Therefore, we investigated whether beta(2)ARs in the human heart activate Galpha(i). By photoaffinity labeling human atrial membranes with [(32)P]azidoanilido-GTP, followed by immunoprecipitation with antibodies specific for Galpha(i), we found that Galpha(i) is activated by stimulation of beta(2)ARs but not of beta(1)ARs. In addition, we found that other Galpha(s)-coupled receptors also couple to Galpha(i), including histamine, serotonin, and glucagon. When coupling of these receptors to Galpha(i) is disrupted by pertussis toxin, their ability to stimulate adenylyl cyclase is enhanced. These data provide the first evidence that beta(2)AR and many other Galpha(s)-coupled receptors in human atrium also couple to Galpha(i) and that abolishing the coupling of these receptors to Galpha(i) increases the receptor-mediated adenylyl cyclase activity. PMID- 11029408 TI - Focal adhesion kinase is involved in angiotensin II-mediated protein synthesis in cultured vascular smooth muscle cells. AB - The rate of vascular smooth muscle cell protein synthesis and cellular hypertrophy in response to angiotensin II (Ang II) is dependent on activation of protein tyrosine kinases (PTKs) and both the extracellular signal-regulated kinase (ERK) 1/2 and p70(S6K) pathways. One potential PTK that may regulate these signaling cascades is focal adhesion kinase (FAK), a nonreceptor PTK associated with focal adhesions. We used an actin depolymerizing agent, cytochalasin D (Cyt D), and a replication-defective adenovirus encoding FAK-related nonkinase (FRNK), an inhibitor of FAK-dependent signaling, as tools to assess whether FAK was upstream of the ERK1/2 and/or the p70(S6K) pathways. Cyt-D reduced basal FAK phosphorylation and blocked Ang II-dependent FAK phosphorylation in a dose dependent manner. Confocal microscopy indicated that Cyt-D induced actin filament disruption and FAK delocalization from focal adhesions. Cyt-D also reduced Ang II induced ERK1/2 activation, but p70(S6K) activation was relatively unaffected. Cyt D reduced basal protein synthetic rate and substantially reduced the Ang II induced increase in protein synthesis. Similarly, FRNK overexpression blocked Ang II-induced FAK phosphorylation and ERK1/2 activation, but not p70(S6K) phosphorylation, and markedly inhibited protein synthesis. This is the first report to demonstrate that FAK is a critical component of the signal transduction pathways that mediate Ang II-induced ERK1/2 activation, c-fos induction, and enhanced protein synthesis in vascular smooth muscle cells. PMID- 11029409 TI - Enhanced Na(+) channel intermediate inactivation in Brugada syndrome. AB - Brugada syndrome is an inherited cardiac disease that causes sudden death related to idiopathic ventricular fibrillation in a structurally normal heart. The disease is characterized by ST-segment elevation in the right precordial ECG leads and is frequently accompanied by an apparent right bundle-branch block. The biophysical properties of the SCN5A mutation T1620M associated with Brugada syndrome were examined for defects in intermediate inactivation (I:(M)), a gating process in Na(+) channels with kinetic features intermediate between fast and slow inactivation. Cultured mammalian cells expressing T1620M Na(+) channels in the presence of the human beta(1) subunit exhibit enhanced intermediate inactivation at both 22 degrees C and 32 degrees C compared with wild-type recombinant human heart Na(+) channels (WT-hH1). Our findings support the hypothesis that Brugada syndrome is caused, in part, by functionally reduced Na(+) current in the myocardium due to an increased proportion of Na(+) channels that enter the I:(M) state. This phenomenon may contribute significantly to arrhythmogenesis in patients with Brugada syndrome. The full text of this article is available at http://www.circresaha.org. PMID- 11029410 TI - Gene fusions. PMID- 11029411 TI - Role of TnrA in nitrogen source-dependent repression of Bacillus subtilis glutamate synthase gene expression. AB - Synthesis of glutamate, the cell's major donor of nitrogen groups and principal anion, occupies a significant fraction of bacterial metabolism. In Bacillus subtilis, the gltAB operon, encoding glutamate synthase, requires a specific positive regulator, GltC, for its expression. In addition, the gltAB operon was shown to be repressed by TnrA, a regulator of several other genes of nitrogen metabolism and active under conditions of ammonium (nitrogen) limitation. TnrA was found to bind directly to a site immediately downstream of the gltAB promoter. As is true for other genes, the activity of TnrA at the gltAB promoter was antagonized by glutamine synthetase under certain growth conditions. PMID- 11029412 TI - Regulation of ferritin-mediated cytoplasmic iron storage by the ferric uptake regulator homolog (Fur) of Helicobacter pylori. AB - Homologs of the ferric uptake regulator Fur and the iron storage protein ferritin play a central role in maintaining iron homeostasis in bacteria. The gastric pathogen Helicobacter pylori contains an iron-induced prokaryotic ferritin (Pfr) which has been shown to be involved in protection against metal toxicity and a Fur homolog which has not been functionally characterized in H. pylori. Analysis of an isogenic fur-negative mutant revealed that H. pylori Fur is required for metal-dependent regulation of ferritin. Iron starvation, as well as medium supplementation with nickel, zinc, copper, and manganese at nontoxic concentrations, repressed synthesis of ferritin in the wild-type strain but not in the H. pylori fur mutant. Fur-mediated regulation of ferritin synthesis occurs at the mRNA level. With respect to the regulation of ferritin expression, Fur behaves like a global metal-dependent repressor which is activated under iron restricted conditions but also responds to different metals. Downregulation of ferritin expression by Fur might secure the availability of free iron in the cytoplasm, especially if iron is scarce or titrated out by other metals. PMID- 11029413 TI - Sequence changes in the ton box region of BtuB affect its transport activities and interaction with TonB protein. AB - Uptake of cobalamins by the transporter protein BtuB in the outer membrane of Escherichia coli requires the proton motive force and the transperiplasmic protein TonB. The Ton box sequence near the amino terminus of BtuB is conserved among all TonB-dependent transporters and is the only known site of mutations that confer a transport-defective phenotype which can be suppressed by certain substitutions at residue 160 in TonB. The crystallographic structures of the TonB dependent transporter FhuA revealed that the region near the Ton box, which itself was not resolved, is exposed to the periplasmic space and undergoes an extensive shift in position upon binding of substrate. Site-directed disulfide bonding in intact cells has been used to show that the Ton box of BtuB and residues around position 160 of TonB approach each other in a highly oriented and specific manner to form BtuB-TonB heterodimers that are stimulated by the presence of transport substrate. Here, replacement of Ton box residues with proline or cysteine revealed that residue side chain recognition is not important for function, although replacement with proline at four of the seven Ton box positions impaired cobalamin transport. The defect in cobalamin utilization resulting from the L8P substitution was suppressed by cysteine substitutions in adjacent residues in BtuB or in TonB. This suppression did not restore active transport of cobalamins but may allow each transporter to function at most once. The uncoupled proline substitutions in BtuB markedly affected the pattern of disulfide bonding to TonB, both increasing the extent of cross-linking and shifting the pairs of residues that can be joined. Cross-linking of BtuB and TonB in the presence of the BtuB V10P substitution became independent of the presence of substrate, indicating an additional distortion of the exposure of the Ton box in the periplasmic space. TonB action thus requires a specific orientation for functional contact with the Ton box, and changes in the conformation of this region block transport by preventing substrate release and repeated transport cycles. PMID- 11029414 TI - Characterization of the distal tail fiber locus and determination of the receptor for phage AR1, which specifically infects Escherichia coli O157:H7. AB - Phage AR1 is similar to phage T4 in several essential genes but differs in host range. AR1 infects various isolates of Escherichia coli O157:H7 but does not infect K-12 strains that are commonly infected by T4. We report here the determinants that confer this infection specificity. In T-even phages, gp37 and gp38 are components of the tail fiber that are critical for phage-host interaction. The counterparts in AR1 may be similarly important and, therefore, were characterized. The AR1 gp37 has a sequence that differs totally from those of T2 and T4, except for a short stretch at the N terminus. The gp38 sequence, however, has some conservation between AR1 and T2 but not between AR1 and T4. The sequences that are most closely related to the AR1 gp37 and gp38 are those of phage Ac3 in the T2 family. To identify the AR1-specific receptor, E. coli O157:H7 was mutated by Tn10 insertion and selected for an AR1-resistant phenotype. A mutant so obtained has an insertion occurring at ompC that encodes an outer membrane porin. To confirm the role of OmpC in the AR1 infection, homologous replacement was used to create an ompC disruption mutant (RM). When RM was complemented with OmpC originated from an O157:H7 strain, but not from K-12, its AR1 susceptibility was fully restored. Our results suggest that the host specificity of AR1 is mediated at least in part through the OmpC molecule. PMID- 11029415 TI - Two-dimensional gel electrophoresis analyses of pH-dependent protein expression in facultatively alkaliphilic Bacillus pseudofirmus OF4 lead to characterization of an S-layer protein with a role in alkaliphily. AB - The large majority of proteins of alkaliphilic Bacillus pseudofirmus OF4 grown at pH 7.5 and 10.5, as studied by two-dimensional gel electrophoresis analyses, did not exhibit significant pH-dependent variation. A new surface layer protein (SlpA) was identified in these studies. Although the prominence of some apparent breakdown products of SlpA in gels from pH 10.5-grown cells led to discovery of the alkaliphile S-layer, the largest and major SlpA forms were present in large amounts in gels from pH 7.5-grown cells as well. slpA RNA abundance was, moreover, unchanged by growth pH. SlpA was similar in size to homologues from nonalkaliphiles but contained fewer Arg and Lys residues. An slpA mutant strain (RG21) lacked an exterior S-layer that was identified in the wild type by electron microscopy. Electrophoretic analysis of whole-cell extracts further indicated the absence of a 90-kDa band in the mutant. This band was prominent in wild-type extracts from both pH 7.5- and 10.5-grown cells. The wild type grew with a shorter lag phase than RG21 at either pH 10.5 or 11 and under either Na(+) replete or suboptimal Na(+) concentrations. The extent of the adaptation deficit increased with pH elevation and suboptimal Na(+). By contrast, the mutant grew with a shorter lag and faster growth rate than the wild type at pH 7. 5 under Na(+)-replete and suboptimal Na(+) conditions, respectively. Logarithmically growing cells of the two strains exhibited no significant differences in growth rate, cytoplasmic pH regulation, starch utilization, motility, Na(+)-dependent transport of alpha-aminoisobutyric acid, or H(+)-dependent synthesis of ATP. However, the capacity for Na(+)-dependent pH homeostasis was diminished in RG21 upon a sudden upward shift of external pH from 8. 5 to 10.5. The energy cost of retaining the SlpA layer at near-neutral pH is apparently adverse, but the constitutive presence of SlpA enhances the capacity of the extremophile to adjust to high pH. PMID- 11029416 TI - Control of lactose transport, beta-galactosidase activity, and glycolysis by CcpA in Streptococcus thermophilus: evidence for carbon catabolite repression by a non phosphoenolpyruvate-dependent phosphotransferase system sugar. AB - Streptococcus thermophilus, unlike many other gram-positive bacteria, prefers lactose over glucose as the primary carbon and energy source. Moreover, lactose is not taken up by a phosphoenolpyruvate-dependent phosphotransferase system (PTS) but by the dedicated transporter LacS. In this paper we show that CcpA plays a crucial role in the fine-tuning of lactose transport, beta-galactosidase (LacZ) activity, and glycolysis to yield optimal glycolytic flux and growth rate. A catabolite-responsive element (cre) was identified in the promoter of the lacSZ operon, indicating a possible role for regulation by CcpA. Transcriptional analysis showed a sevenfold relief of repression in the absence of a functional CcpA when cells were grown on lactose. This CcpA-mediated repression of lacSZ transcription did not occur in wild-type cells during growth on galactose, taken up by the same LacS transport system. Lactose transport during fermentation was increased significantly in strains carrying a disrupted ccpA gene. Moreover, a ccpA disruption strain was found to release substantial amounts of glucose into the medium when grown on lactose. Transcriptional analysis of the ldh gene showed that expression was induced twofold during growth on lactose compared to glucose or galactose, in a CcpA-dependent manner. A reduced rate of glycolysis concomitant with an increased lactose transport rate could explain the observed expulsion of glucose in a ccpA disruption mutant. We propose that CcpA in S. thermophilus acts as a catabolic regulator during growth on the preferred non-PTS sugar lactose. In contrast to other bacteria, S. thermophilus possesses an overcapacity for lactose uptake that is repressed by CcpA to match the rate limiting glycolytic flux. PMID- 11029417 TI - Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. AB - We describe swarming in Pseudomonas aeruginosa as a third mode of surface translocation in addition to the previously described swimming and twitching motilities. Swarming in P. aeruginosa is induced on semisolid surfaces (0.5 to 0.7% agar) under conditions of nitrogen limitation and in response to certain amino acids. Glutamate, aspartate, histidine, or proline, when provided as the sole source of nitrogen, induced swarming, while arginine, asparagine, and glutamine, among other amino acids, did not sustain swarming. Cells from the edge of the swarm were about twice as long as cells from the swarm center. In both instances, bacteria possessing two polar flagella were observed by light and electron microscopy. While a fliC mutant of P. aeruginosa displayed slightly diminished swarming, a pilR and a pilA mutant, both deficient in type IV pili, were unable to swarm. Furthermore, cells with mutations in the las cell-to-cell signaling system showed diminished swarming behavior, while rhl mutants were completely unable to swarm. Evidence is presented for rhamnolipids being the actual surfactant involved in swarming motility, which explains the involvement of the cell-to-cell signaling circuitry of P. aeruginosa in this type of surface motility. PMID- 11029418 TI - Characterization of the hydrogen-deuterium exchange activities of the energy transducing HupSL hydrogenase and H(2)-signaling HupUV hydrogenase in Rhodobacter capsulatus. AB - Rhodobacter capsulatus synthesizes two homologous protein complexes capable of activating molecular H(2), a membrane-bound [NiFe] hydrogenase (HupSL) linked to the respiratory chain, and an H(2) sensor encoded by the hupUV genes. The activities of hydrogen-deuterium (H-D) exchange catalyzed by the hupSL-encoded and the hupUV-encoded enzymes in the presence of D(2) and H(2)O were studied comparatively. Whereas HupSL is in the membranes, HupUV activity was localized in the soluble cytoplasmic fraction. Since the hydrogenase gene cluster of R. capsulatus contains a gene homologous to hoxH, which encodes the large subunit of NAD-linked tetrameric soluble hydrogenases, the chromosomal hoxH gene was inactivated and hoxH mutants were used to demonstrate the H-D exchange activity of the cytoplasmic HupUV protein complex. The H-D exchange reaction catalyzed by HupSL hydrogenase was maximal at pH 4. 5 and inhibited by acetylene and oxygen, whereas the H-D exchange catalyzed by the HupUV protein complex was insensitive to acetylene and oxygen and did not vary significantly between pH 4 and pH 11. Based on these properties, the product of the accessory hypD gene was shown to be necessary for the synthesis of active HupUV enzyme. The kinetics of HD and H(2) formed in exchange with D(2) by HupUV point to a restricted access of protons and gasses to the active site. Measurement of concentration changes in D(2), HD, and H(2) by mass spectrometry showed that, besides the H-D exchange reaction, HupUV oxidized H(2) with benzyl viologen, produced H(2) with reduced methyl viologen, and demonstrated true hydrogenase activity. Therefore, not only with respect to its H(2) signaling function in the cell, but also to its catalytic properties, the HupUV enzyme represents a distinct class of hydrogenases. PMID- 11029419 TI - The Haemophilus influenzae Hia adhesin is an autotransporter protein that remains uncleaved at the C terminus and fully cell associated. AB - Nontypeable Haemophilus influenzae is a gram-negative commensal organism that is commonly associated with localized respiratory tract disease. The pathogenesis of disease begins with colonization of the nasopharynx, a process that likely depends on bacterial adherence to respiratory epithelial cells. Hia is the major adhesin expressed by a subset of nontypeable H. influenzae strains and promotes efficient adherence to a variety of human epithelial cell lines. Based on previous work, Hia is transported to the surface of Escherichia coli transformants and is capable of mediating E. coli adherence without the assistance of other H. influenzae proteins. In the present study, we examined the mechanism of Hia secretion. PhoA fusions, deletional mutagenesis, and N-terminal amino acid sequencing established that the signal for Hia export from the cytoplasm resides in the first 49 amino acids, including a 24-amino-acid stretch with striking similarity to the N terminus of a number of proteins belonging to the autotransporter family. Immunoelectron microscopy demonstrated that the Hia internal region defined by amino acids 221 to 779 is exposed on the bacterial surface. Secondary-structure analysis predicted that the C terminus of Hia forms a beta-barrel with a central hydrophilic channel, and site-specific mutagenesis and fusion protein analysis demonstrated that the C terminus targets Hia to the outer membrane and functions as an outer membrane translocator, analogous to observations with autotransporter proteins. In contrast to typical autotransporter proteins, Hia undergoes no cleavage between the internal and C terminal domains and remains fully cell associated. Together, these results suggest that Hia is the prototype of an important subfamily of autotransporter proteins. PMID- 11029421 TI - RNA polymerases from Bacillus subtilis and Escherichia coli differ in recognition of regulatory signals in vitro. AB - Adaptation of bacterial cells to diverse habitats relies on the ability of RNA polymerase to respond to various regulatory signals. Some of these signals are conserved throughout evolution, whereas others are species specific. In this study we present a comprehensive comparative analysis of RNA polymerases from two distantly related bacterial species, Escherichia coli and Bacillus subtilis, using a panel of in vitro transcription assays. We found substantial species specific differences in the ability of these enzymes to escape from the promoter and to recognize certain types of elongation signals. Both enzymes responded similarly to other pause and termination signals and to the general E. coli elongation factors NusA and GreA. We also demonstrate that, although promoter recognition depends largely on the sigma subunit, promoter discrimination exhibited in species-specific fashion by both RNA polymerases resides in the core enzyme. We hypothesize that differences in signal recognition are due to the changes in contacts made between the beta and beta' subunits and the downstream DNA duplex. PMID- 11029420 TI - Incompatibility protein IncC and global regulator KorB interact in active partition of promiscuous plasmid RK2. AB - Replication of the broad-host-range, IncPalpha plasmid RK2 requires two plasmid loci: trfA, the replication initiator gene, and oriV, the origin of replication. While these determinants are sufficient for replication in a wide variety of bacteria, they do not confer the stable maintenance of parental RK2 observed in its hosts. The product of the incC gene has been proposed to function in the stable maintenance of RK2 because of its relatedness to the ParA family of ATPases, some of which are known to be involved in the active partition of plasmid and chromosomal DNA. Here we show that IncC has the properties expected of a component of an active partition system. The smaller polypeptide product of incC (IncC2) exhibits a strong, replicon-independent incompatibility phenotype with RK2. This incompatibility phenotype requires the global transcriptional repressor, KorB, and the target for incC-mediated incompatibility is a KorB binding site (O(B)). We found that KorB and IncC interact in vivo by using the yeast two-hybrid system and in vitro by using partially purified proteins. Elevated expression of the incC and korB genes individually has no obvious effect on Escherichia coli cell growth, but their simultaneous overexpression is toxic, indicating a possible interaction of IncC-KorB complexes with a vital host target. A region of RK2 bearing incC, korB, and multiple KorB-binding sites is able to stabilize an unstable, heterologous plasmid in an incC-dependent manner. Finally, elevated levels of IncC2 cause RK2 to aggregate, indicating a possible role for IncC in plasmid pairing. These findings demonstrate that IncC, KorB, and at least one KorB-binding site are components of an active partition system for the promiscuous plasmid RK2. PMID- 11029422 TI - Mre11 and Rad50 from Pyrococcus furiosus: cloning and biochemical characterization reveal an evolutionarily conserved multiprotein machine. AB - The processing of DNA double-strand breaks is a critical event in nucleic acid metabolism. This is evidenced by the severity of phenotypes associated with deficiencies in this process in multiple organisms. The core component involved in double-strand break repair in eukaryotic cells is the Mre11-Rad50 protein complex, which includes a third protein, p95, in humans and Xrs2 in yeasts. Homologues of Mre11 and Rad50 have been identified in all kingdoms of life, while the Nbs1 protein family is found only in eukaryotes. In eukaryotes the Mre11 Rad50 complex has nuclease activity that is modulated by the addition of ATP. We have isolated the Mre11 and Rad50 homologues from the thermophilic archaeon Pyrococcus furiosus and demonstrate that the two proteins exist in a large, heat stable complex that possesses single-strand endonuclease activity and ATP dependent double-strand-specific exonuclease activity. These findings verify the identification of the P. furiosus Rad50 and Mre11 homologues and demonstrate that functional homologues with similar biochemical properties exist in all kingdoms of life. PMID- 11029423 TI - Energy taxis is the dominant behavior in Azospirillum brasilense. AB - Energy taxis encompasses aerotaxis, phototaxis, redox taxis, taxis to alternative electron acceptors, and chemotaxis to oxidizable substrates. The signal for this type of behavior is originated within the electron transport system. Energy taxis was demonstrated, as a part of an overall behavior, in several microbial species, but it did not appear as the dominant determinant in any of them. In this study, we show that most behavioral responses proceed through this mechanism in the alpha-proteobacterium Azospirillum brasilense. First, chemotaxis to most chemoeffectors typical of the azospirilla habitat was found to be metabolism dependent and required a functional electron transport system. Second, other energy-related responses, such as aerotaxis, redox taxis, and taxis to alternative electron acceptors, were found in A. brasilense. Finally, a mutant lacking a cytochrome c oxidase of the cbb(3) type was affected in chemotaxis, redox taxis, and aerotaxis. Altogether, the results indicate that behavioral responses to most stimuli in A. brasilense are triggered by changes in the electron transport system. PMID- 11029424 TI - Aerobic activity of Escherichia coli alcohol dehydrogenase is determined by a single amino acid. AB - Expression of the alcohol dehydrogenase gene, adhE, in Escherichia coli is anaerobically regulated at both the transcriptional and the translational levels. To study the AdhE protein, the adhE(+) structural gene was cloned into expression vectors under the control of the lacZ and trp(c) promoters. Wild-type AdhE protein produced under aerobic conditions from these constructs was inactive. Constitutive mutants (adhC) that produced high levels of AdhE under both aerobic and anaerobic conditions were previously isolated. When only the adhE structural gene from one of the adhC mutants was cloned into expression vectors, highly functional AdhE protein was isolated under both aerobic and anaerobic conditions. Sequence analysis revealed that the adhE gene from the adhC mutant contained two mutations resulting in two amino acid substitutions, Ala267Thr and Glu568Lys. Thus, adhC strains contain a promoter mutation and two mutations in the structural gene. The mutant structural gene from adhC strains was designated adhE*. Fragment exchange experiments revealed that the substitution responsible for aerobic expression in the adhE* clones is Glu568Lys. Genetic selection and site-directed mutagenesis experiments showed that virtually any amino acid substitution for Glu568 produced AdhE that was active under both aerobic and anaerobic conditions. These findings suggest that adhE expression is also regulated posttranslationally and that strict regulation of alcohol dehydrogenase activity in E. coli is physiologically significant. PMID- 11029425 TI - Defects in D-alanyl-lipoteichoic acid synthesis in Streptococcus mutans results in acid sensitivity. AB - In the cariogenic organism, Streptococcus mutans, low pH induces an acid tolerance response (ATR). To identify acid-regulated proteins comprising the ATR, transposon mutagenesis with the thermosensitive plasmid pGh9:ISS1 was used to produce clones that were able to grow at neutral pH, but not in medium at pH 5.0. Sequence analysis of one mutant (IS1A) indicated that transposition had created a 6.3-kb deletion, one end of which was in dltB of the dlt operon encoding four proteins (DltA-DltD) involved in the synthesis of D-alanyl-lipoteichoic acid. Inactivation of the dltC gene, encoding the D-alanyl carrier protein (Dcp), resulted in the generation of the acid-sensitive mutant, BH97LC. Compared to the wild-type strain, LT11, the mutant exhibited a threefold-longer doubling time and a 33% lower growth yield. In addition, it was unable to initiate growth below pH 6.5 and unadapted cells were unable to survive a 3-h exposure in medium buffered at pH 3.5, while a pH of 3.0 was required to kill the wild type in the same time period. Also, induction of the ATR in BH97LC, as measured by the number of survivors at a pH killing unadapted cells, was 3 to 4 orders of magnitude lower than that exhibited by the wild type. While the LTA of both strains contained a similar average number of glycerolphosphate residues, permeabilized cells of BH97LC did not incorporate D-[(14)C]alanine into this amphiphile. This defect was correlated with the deficiency of Dcp. Chemical analysis of the LTA purified from the mutant confirmed the absence of D-alanine-esters. Electron micrographs showed that BH97LC is characterized by unequal polar caps and is devoid of a fibrous extracellular matrix present on the surface of the wild-type cells. Proton permeability assays revealed that the mutant was more permeable to protons than the wild type. This observation suggests a mechanism for the loss of the characteristic acid tolerance response in S. mutans. PMID- 11029426 TI - Sequence of the genome of the temperate, serotype-converting, Pseudomonas aeruginosa bacteriophage D3. AB - Temperate bacteriophage D3, a member of the virus family Siphoviridae, is responsible for serotype conversion in its host, Pseudomonas aeruginosa. The complete sequence of the double-stranded DNA genome has been determined. The 56,426 bp contains 90 putative open reading frames (ORFs) and four genes specifying tRNAs. The latter are specific for methionine (AUG), glycine (GGA), asparagine (AAC), and threonine (ACA). The tRNAs may function in the translation of certain highly expressed proteins from this relatively AT-rich genome. D3 proteins which exhibited a high degree of sequence similarity to previously characterized phage proteins included the portal, major head, tail, and tail tape measure proteins, endolysin, integrase, helicase, and NinG. The layout of genes was reminiscent of lambdoid phages, with the exception of the placement of the endolysin gene, which parenthetically also lacked a cognate holin. The greatest sequence similarity was found in the morphogenesis genes to coliphages HK022 and HK97. Among the ORFs was discovered the gene encoding the fucosamine O-acetylase, which is in part responsible for the serotype conversion events. PMID- 11029427 TI - Dimerization between the holin and holin inhibitor of phage lambda. AB - Holins are integral membrane proteins that control the access of phage-encoded muralytic enzymes, or endolysins, to the cell wall by the sudden formation of an uncharacterized homo-oligomeric lesion, or hole, in the membrane, at a precisely defined time. The timing of lambda-infected cell lysis depends solely on the 107 codon S gene, which encodes two proteins, S105 and S107, which are the holin and holin inhibitor, respectively. Here we report the results of biochemical and genetic studies on the interaction between the holin and the holin inhibitor. A unique cysteine at position 51, in the middle of the second transmembrane domain, is shown to cause the formation of disulfide-linked dimers during detergent membrane extraction. Forced oxidation of membranes containing S molecules also results in the formation of covalently linked dimers. This technique is used to demonstrate efficient dimeric interactions between S105 and S107. These results, coupled with the previous finding that the timing of lysis depends on the excess of the amount of S105 over S107, suggest a model in which the inhibitor functions by titrating out the effector in a stoichiometric fashion. This provides a basis for understanding two evolutionary advantages provided by the inhibitor system, in which the production of the inhibitor not only causes a delay in the timing of lysis, allowing the assembly of more virions, but also increases effective hole formation after triggering. PMID- 11029428 TI - Genetic and biochemical analysis of dimer and oligomer interactions of the lambda S holin. AB - Bacteriophage lambda uses a holin-endolysin system for host cell lysis. R, the endolysin, has muralytic activity. S, the holin, is a small membrane protein that permeabilizes the inner membrane at a precisely scheduled time after infection and allows the endolysin access to its substrate, resulting in host cell lysis. lambda S has a single cysteine at position 51 that can be replaced by a serine without loss of the holin function. A collection of 27 single-cysteine products of alleles created from lambda S(C51S) were tested for holin function. Most of the single-cysteine variants retained the ability to support lysis. Mutations with the most defective phenotype clustered in the first two hydrophobic transmembrane domains. Several lines of evidence indicate that S forms an oligomeric structure in the inner membrane. Here we show that oligomerization does not depend on disulfide bridge formation, since the cysteineless S(C51S) (i) is functional as a holin and (ii) shows the same oligomerization pattern as the parental S protein. In contrast, the lysis-defective S(A52V) mutant dimerizes but does not form cross-linkable oligomers. Again, dimerization does not depend on the natural cysteine, since the cysteineless lysis-defective S(A52V/C51S) is found in dimers after treatment of the membrane with a cross-linking agent. Furthermore, under oxidative conditions, dimerization via the natural cysteine is very efficient for S(A52V). Both S(A52V) (dominant negative) and S(A48V) (antidominant) interact with the parental S protein, as judged by oxidative disulfide bridge formation. Thus, productive and unproductive heterodimer formation between the parental protein and the mutants S(A52V) and S(A48V), respectively, may account for the dominant and antidominant lysis phenotypes. Examination of oxidative dimer formation between S variants with single cysteines in the hydrophobic core of the second membrane-spanning domain revealed that positions 48 and 51 are on a dimer interface. These results are discussed in terms of a three-step model leading to S-dependent hole formation in the inner membrane. PMID- 11029429 TI - The mammalian neuroendocrine hormone norepinephrine supplies iron for bacterial growth in the presence of transferrin or lactoferrin. AB - Norepinephrine stimulates the growth of a range of bacterial species in nutritionally poor SAPI minimal salts medium containing 30% serum. Addition of size-fractionated serum components to SAPI medium indicated that transferrin was required for norepinephrine stimulation of growth of Escherichia coli. Since bacteriostasis by serum is primarily due to the iron-withholding capacity of transferrin, we considered the possibility that norepinephrine can overcome this effect by supplying transferrin-bound iron for growth. Incubation with concentrations of norepinephrine that stimulated bacterial growth in serum-SAPI medium resulted in loss of bound iron from iron-saturated transferrin, as indicated by the appearance of monoferric and apo- isoforms upon electrophoresis in denaturing gels. Norepinephrine also caused the loss of iron from lactoferrin. The pharmacologically inactive metabolite norepinephrine 3-O-sulfate, by contrast, did not result in iron loss from transferrin or lactoferrin and did not stimulate bacterial growth in serum-SAPI medium. Norepinephrine formed stable complexes with transferrin, lactoferrin, and serum albumin. Norepinephrine transferrin and norepinephrine-lactoferrin complexes, but not norepinephrine apotransferrin or norepinephrine-albumin complexes, stimulated bacterial growth in serum-SAPI medium in the absence of additional norepinephrine. Norepinephrine stimulated growth in medium containing (55)Fe complexed with transferrin or lactoferrin resulted in uptake of radioactivity by bacterial cells. Moreover, norepinephrine-stimulated growth in medium containing [(3)H]norepinephrine indicated concomitant uptake of norepinephrine. In each case, addition of excess iron did not affect growth but significantly reduced levels of radioactivity ((55)Fe or (3)H) associated with bacterial cells. A role for catecholamine mediated iron supply in the pathophysiology of infectious diseases is proposed. PMID- 11029430 TI - Catabolite repression and induction of the Mg(2+)-citrate transporter CitM of Bacillus subtilis. AB - In Bacillus subtilis the citM gene encodes the Mg(2+)-citrate transporter. A target site for carbon catabolite repression (cre site) is located upstream of citM. Fusions of the citM promoter region, including the cre sequence, to the beta-galactosidase reporter gene were constructed and integrated into the amyE site of B. subtilis to study catabolic effects on citM expression. In parallel with beta-galactosidase activity, the uptake of Ni(2+)-citrate in whole cells was measured to correlate citM promoter activity with the enzymatic activity of the CitM protein. In minimal media, CitM was only expressed when citrate was present. The presence of glucose in the medium completely repressed citM expression; repression was also observed in media containing glycerol, inositol, or succinate glutamate. Studies with B. subtilis mutants defective in the catabolite repression components HPr, Crh, and CcpA showed that the repression exerted by all these medium components was mediated via the carbon catabolite repression system. During growth on inositol and succinate, the presence of glutamate strongly potentiated the repression of citM expression by glucose. A reasonable correlation between citM promoter activity and CitM transport activity was observed in this study, indicating that the Mg(2+)-citrate uptake activity of B. subtilis is mainly regulated at the transcriptional level. PMID- 11029431 TI - Escherichia coli DNA polymerase III tau- and gamma-subunit conserved residues required for activity in vivo and in vitro. AB - The Escherichia coli DNA polymerase III tau and gamma subunits are single-strand DNA-dependent ATPases (the latter requires the delta and delta' subunits for significant ATPase activity) involved in loading processivity clamp beta. They are homologous to clamp-loading proteins of many organisms from phages to humans. Alignment of 27 prokaryotic tau/gamma homologs and 1 eukaryotic tau/gamma homolog has refined the sequences of nine previously defined identity and functional motifs. Mutational analysis has defined highly conserved residues required for activity in vivo and in vitro. Specifically, mutations introduced into highly conserved residues within three of those motifs, the P loop, the DExx region, and the SRC region, inactivated complementing activity in vivo and clamp loading in vitro and reduced ATPase catalytic efficiency in vitro. Mutation of a highly conserved residue within a fourth motif, VIc, inactivated clamp-loading activity and reduced ATPase activity in vitro, but the mutant gene, on a multicopy plasmid, retained complementing activity in vivo and the mutant gene also supported apparently normal replication and growth as a haploid, chromosomal allele. PMID- 11029432 TI - Cloning and characterization of the str operon and elongation factor Tu expression in Bacillus stearothermophilus. AB - The complete primary structure of the str operon of Bacillus stearothermophilus was determined. It was established that the operon is a five-gene transcriptional unit: 5'-ybxF (unknown function; homology to eukaryotic ribosomal protein L30) rpsL (S12)-rpsG (S7)-fus (elongation factor G [EF-G])-tuf (elongation factor Tu [EF-Tu])-3'. The main operon promoter (strp) was mapped upstream of ybxF, and its strength was compared with the strength of the tuf-specific promoter (tufp) located in the fus-tuf intergenic region. The strength of the tufp region to initiate transcription is about 20-fold higher than that of the strp region, as determined in chloramphenicol acetyltransferase assays. Deletion mapping experiments revealed that the different strengths of the promoters are the consequence of a combined effect of oppositely acting cis elements, identified upstream of strp (an inhibitory region) and tufp (a stimulatory A/T-rich block). Our results suggest that the oppositely adjusted core promoters significantly contribute to the differential expression of the str operon genes, as monitored by the expression of EF-Tu and EF-G. PMID- 11029433 TI - Characterization of the genes for two protocatechuate 3, 4-dioxygenases from the 4-sulfocatechol-degrading bacterium Agrobacterium radiobacter strain S2. AB - The genes for two different protocatechuate 3,4-dioxygenases (P34Os) were cloned from the 4-sulfocatechol-degrading bacterium Agrobacterium radiobacter strain S2 (DSMZ 5681). The pcaH1G1 genes encoded a P34O (P34O-I) which oxidized protocatechuate but not 4-sulfocatechol. These genes were part of a protocatechuate-degradative operon which strongly resembled the isofunctional operon from the protocatechuate-degrading strain Agrobacterium tumefaciens A348 described previously by D. Parke (FEMS Microbiol. Lett. 146:3-12, 1997). The second P34O (P34O-II), encoded by the pcaH2G2 genes, was functionally expressed and shown to convert protocatechuate and 4-sulfocatechol. A comparison of the deduced amino acid sequences of PcaH-I and PcaH-II, and of PcaG-I and PcaG-II, with each other and with the corresponding sequences from the P34Os, from other bacterial genera suggested that the genes for the P34O-II were obtained by strain S2 by lateral gene transfer. The genes encoding the P34O-II were found in a putative operon together with two genes which, according to sequence alignments, encoded transport proteins. Further downstream from this putative operon, two open reading frames which code for a putative regulator protein of the IclR family and a putative 3-carboxymuconate cycloisomerase were identified. PMID- 11029434 TI - Discovery, purification, and characterization of a temperate transducing bacteriophage for Bordetella avium. AB - We discovered and characterized a temperate transducing bacteriophage (Ba1) for the avian respiratory pathogen Bordetella avium. Ba1 was initially identified along with one other phage (Ba2) following screening of four strains of B. avium for lysogeny. Of the two phage, only Ba1 showed the ability to transduce via an allelic replacement mechanism and was studied further. With regard to host range, Ba1 grew on six of nine clinical isolates of B. avium but failed to grow on any tested strains of Bordetella bronchiseptica, Bordetella hinzii, Bordetella pertussis, or Bordetella parapertussis. Ba1 was purified by CsCl gradient centrifugation and was found to have an icosahedral head that contained a linear genome of approximately 46.5 kb (contour length) of double-stranded DNA and a contractile, sheathed tail. Ba1 readily lysogenized our laboratory B. avium strain (197N), and the prophage state was stable for at least 25 generations in the absence of external infection. DNA hybridization studies indicated the prophage was integrated at a preferred site on both the host and phage replicons. Ba1 transduced five distinctly different insertion mutations, suggesting that transduction was generalized. Transduction frequencies ranged from approximately 2 x 10(-7) to 1 x 10(-8) transductants/PFU depending upon the marker being transduced. UV irradiation of transducing lysates markedly improved transduction frequency and reduced the number of transductants that were lysogenized during the transduction process. Ba1 may prove to be a useful genetic tool for studying B. avium virulence factors. PMID- 11029435 TI - Regulation of the rulAB mutagenic DNA repair operon of Pseudomonas syringae by UV B (290 to 320 nanometers) radiation and analysis of rulAB-mediated mutability in vitro and in planta. AB - The effects of the rulAB operon of Pseudomonas syringae on mutagenic DNA repair and the transcriptional regulation of rulAB following irradiation with UV-B wavelengths were determined. For a rulB::Km insertional mutant constructed in P. syringae pv. syringae B86-17, sensitivity to UV-B irradiation increased and UV mutability decreased by 12- to 14-fold. rulAB-induced UV mutability was also tracked in phyllosphere populations of B86-17 for up to 5 days following plant inoculation. UV mutability to rifampin resistance (Rif(r)) was detected at all sampling points at levels which were significantly greater than in nonirradiated controls. In P. aeruginosa PAO1, the cloned rulAB determinant on pJJK17 conferred a 30-fold increase in survival and a 200-fold increase in mutability following a UV-B dose of 1,900 J m(-2). In comparative studies using defined genetic constructs, we determined that rulAB restored mutability to the Escherichia coli umuDC deletion mutant RW120 at a level between those of its homologs mucAB and umuDC. Analyses using a rulAB::inaZ transcriptional fusion in Pseudomonas fluorescens Pf5 showed that rulAB was rapidly induced after UV-B irradiation, with expression levels peaking at 4 h. At the highest UV-B dose administered, transcriptional activity of the rulAB promoter was elevated as much as 261-fold compared to that of a nonirradiated control. The importance of rulAB for survival of P. syringae in its phyllosphere habitat, coupled with its wide distribution among a broad range of P. syringae genotypes, suggests that this determinant would be appropriate for continued investigations into the ecological ramifications of mutagenic DNA repair. PMID- 11029436 TI - Positive selection for mutations affecting bioconversion of aromatic compounds in Agrobacterium tumefaciens: analysis of spontaneous mutations in the protocatechuate 3,4-dioxygenase gene. AB - A positive selection method for mutations affecting bioconversion of aromatic compounds was applied to a mutant strain of Agrobacterium tumefaciens A348. The nucleotide sequence of the A348 pcaHGB genes, which encode protocatechuate 3,4 dioxygenase (PcaHG) and beta-carboxy-cis,cis-muconate cycloisomerase (PcaB) for the first two steps in catabolism of the diphenolic protocatechuate, was determined. An omega element was introduced into the pcaB gene of A348, creating strain ADO2077. In the presence of phenolic compounds that can serve as carbon sources, growth of ADO2077 is inhibited due to accumulation of the tricarboxylate intermediate. The toxic effect, previously described for Acinetobacter sp., affords a powerful selection for suppressor mutations in genes required for upstream catabolic steps. By monitoring loss of the marker in pcaB, it was possible to determine that the formation of deletions was minimal compared to results obtained with Acinetobacter sp. Thus, the tricarboxylic acid trick in and of itself does not appear to select for large deletion mutations. The power of the selection was demonstrated by targeting the pcaHG genes of A. tumefaciens for spontaneous mutation. Sixteen strains carrying putative second-site mutations in pcaH or -G were subjected to sequence analysis. All single-site events, their mutations revealed no particular bias toward multibase deletions or unusual patterns: five (-1) frameshifts, one (+1) frameshift, one tandem duplication of 88 bp, one deletion of 92 bp, one nonsense mutation, and seven missense mutations. PcaHG is considered to be the prototypical ferric intradiol dioxygenase. The missense mutations served to corroborate the significance of active site amino acid residues deduced from crystal structures of PcaHG from Pseudomonas putida and Acinetobacter sp. as well as of residues in other parts of the enzyme. PMID- 11029437 TI - Inhibitory effect of heterologous ribosome recycling factor on growth of Escherichia coli. AB - Ribosome recycling factor (RRF) of Thermotoga maritima was expressed in Escherichia coli from the cloned T. maritima RRF gene and purified. Expression of T. maritima RRF inhibited growth of the E. coli host in a dose-dependent manner, an effect counteracted by the overexpression of E. coli RRF. T. maritima RRF also inhibited the E. coli RRF reaction in vitro. Genes encoding RRFs from Streptococcus pneumoniae and Helicobacter pylori have been cloned, and they also impair growth of E. coli, although the inhibitory effect of these RRFs was less pronounced than that of T. maritima RRF. The amino acid sequence at positions 57 to 62, 74 to 78, 118 to 122, 154 to 160, and 172 to 176 in T. maritima RRF differed totally from that of E. coli RRF. This suggests that these regions are important for the inhibitory effect of heterologous RRF. We further suggest that bending and stretching of the RRF molecule at the hinge between two domains may be critical for RRF activity and therefore responsible for T. maritima RRF inhibition of the E. coli RRF reaction. PMID- 11029438 TI - A sheep in wolf's clothing: Listeria innocua strains with teichoic acid associated surface antigens and genes characteristic of Listeria monocytogenes serogroup 4. AB - Listeria monocytogenes serotype 4b has been implicated in numerous food-borne epidemics and in a substantial fraction of sporadic listeriosis. A unique lineage of the nonpathogenic species Listeria innocua was found to express teichoic acid associated surface antigens that were otherwise expressed only by L. monocytogenes of serotype 4b and the rare serotypes 4d and 4e. These L. innocua strains were also found to harbor sequences homologous to the gene gtcA, which has been shown to be essential for teichoic acid glycosylation in L. monocytogenes serotype 4b. Transposon mutagenesis and genetic studies revealed that the gtcA gene identified in this lineage of L. innocua was functional in serotype 4b-like glycosylation of the teichoic acids of these organisms. The genomic organization of the gtcA region was conserved between this lineage of L. innocua and L. monocytogenes serotype 4b. Our data are in agreement with the hypothesis that, in this lineage of L. innocua, gtcA was acquired by lateral transfer from L. monocytogenes serogroup 4. The high degree of nucleotide sequence conservation in the gtcA sequences suggests that such transfer was relatively recent. Transfer events of this type may alter the surface antigenic properties of L. innocua and may eventually lead to evolution of novel pathogenic lineages through additional acquisition of genes from virulent listeriae. PMID- 11029439 TI - Nonspecific adherence by Actinobacillus actinomycetemcomitans requires genes widespread in bacteria and archaea. AB - The gram-negative coccobacillus, Actinobacillus actinomycetemcomitans, is the putative agent for localized juvenile periodontitis, a particularly destructive form of periodontal disease in adolescents. This bacterium has also been isolated from a variety of other infections, notably endocarditis. Fresh clinical isolates of A. actinomycetemcomitans form tenacious biofilms, a property likely to be critical for colonization of teeth and other surfaces. Here we report the identification of a locus of seven genes required for nonspecific adherence of A. actinomycetemcomitans to surfaces. The recently developed transposon IS903phikan was used to isolate mutants of the rough clinical isolate CU1000 that are defective in tight adherence to surfaces (Tad(-)). Unlike wild-type cells, Tad(-) mutant cells adhere poorly to surfaces, fail to form large autoaggregates, and lack long, bundled fibrils. Nucleotide sequencing and genetic complementation analysis revealed a 6.7-kb region of the genome with seven adjacent genes (tadABCDEFG) required for tight adherence. The predicted TadA polypeptide is similar to VirB11, an ATPase involved in macromolecular transport. The predicted amino acid sequences of the other Tad polypeptides indicate membrane localization but no obvious functions. We suggest that the tad genes are involved in secretion of factors required for tight adherence of A. actinomycetemcomitans. Remarkably, complete and highly conserved tad gene clusters are present in the genomes of the bubonic plague bacillus Yersinia pestis and the human and animal pathogen Pasteurella multocida. Partial tad loci also occur in strikingly diverse Bacteria and Archaea. Our results show that the tad genes are required for tight adherence of A. actinomycetemcomitans to surfaces and are therefore likely to be essential for colonization and pathogenesis. The occurrence of similar genes in a wide array of microorganisms indicates that they have important functions. We propose that tad-like genes have a significant role in microbial colonization. PMID- 11029440 TI - A genetic mechanism for deletion of the ser2 gene cluster and formation of rough morphological variants of Mycobacterium avium. AB - A major phenotypic trait of the Mycobacterium avium complex is the ability to produce rough and smooth colony variants. The chemical basis of this morphological variation is the loss of an antigenic surface structure, termed glycopeptidolipid (GPL), by rough variants. Using M. avium serovar 2 strain 2151 as a model system, this laboratory previously reported that rough variants arise via the deletion of large genomic regions encoding GPL biosynthesis. One such deletion encompasses the gene cluster (ser2) responsible for production of the serovar 2 GPL haptenic oligosaccharide. In this study, nucleotide sequencing revealed that both ends of the ser2 gene cluster are flanked by a novel insertion sequence (IS1601) oriented as direct repeats. Detailed analyses of the site of deletion in the genome of M. avium 2151 Rg-1 demonstrated that a single copy of IS1601 remained and that the ser2 gene cluster was deleted by homologous recombination. This same deletion pattern was observed for 10 out of 15 rough colony variants tested. Additionally, these studies revealed that IS1601 contains portions of three independent insertion sequences. This report is the first to define the precise genetic basis of colony variation in Mycobacterium spp. and provides further evidence that homologous recombination between insertion sequence elements can be a primary determinant of genome plasticity in these bacteria. PMID- 11029441 TI - Acquisition of the rfb-gnd cluster in evolution of Escherichia coli O55 and O157. AB - The rfb region specifies the structure of lipopolysaccharide side chains that comprise the diverse gram-negative bacterial somatic (O) antigens. The rfb locus is adjacent to gnd, which is a polymorphic gene encoding 6-phosphogluconate dehydrogenase. To determine if rfb and gnd cotransfer, we sequenced gnd in five O55 and 13 O157 strains of Escherichia coli. E. coli O157:H7 has a gnd allele (allele A) that is only 82% identical to the gnd allele (allele D) of closely related E. coli O55:H7. In contrast, gnd alleles of E. coli O55 in distant lineages are >99.9% identical to gnd allele D. Though gnd alleles B and C in E. coli O157 that are distantly related to E. coli O157:H7 are more similar to allele A than to allele D, there are nucleotide differences at 4 to 6% of their sites. Alleles B and C can be found in E. coli O157 in different lineages, but we have found allele A only in E. coli O157 belonging to the DEC5 lineage. DNA 3' to the O55 gnd allele in diverse E. coli lineages has sequences homologous to tnpA of the Salmonella enterica serovar Typhimurium IS200 element, E. coli Rhs elements (including an H-rpt gene), and portions of the O111 and O157 rfb regions. We conclude that rfb and gnd cotransferred into E. coli O55 and O157 in widely separated lineages and that recombination was responsible for recent antigenic shifts in the emergence of pathogenic E. coli O55 and O157. PMID- 11029442 TI - Gene expression analysis of the Streptococcus pneumoniae competence regulons by use of DNA microarrays. AB - Competence for genetic transformation in Streptococcus pneumoniae is coordinated by the competence-stimulating peptide (CSP), which induces a sudden and transient appearance of competence during exponential growth in vitro. Models of this quorum-sensing mechanism have proposed sequential expression of several regulatory genes followed by induction of target genes encoding DNA-processing pathway proteins. Although many genes required for transformation are known to be expressed only in response to CSP, the relative timing of their expression has not been established. Overlapping expression patterns for the genes cinA and comD (G. Alloing, B. Martin, C. Granadel, and J. P. Claverys, Mol. Microbiol. 29:75 83, 1998) suggest that at least two distinct regulatory mechanisms may underlie the competence cycle. DNA microarrays were used to estimate mRNA levels for all known competence operons during induction of competence by CSP. The known competence regulatory operons, comAB, comCDE, and comX, exhibited a low or zero initial (uninduced) signal, strongly increased expression during the period between 5 and 12 min after CSP addition, and a decrease nearly to original values by 15 min after initiation of exposure to CSP. The remaining competence genes displayed a similar expression pattern, but with an additional delay of approximately 5 min. In a mutant defective in ComX, which may act as an alternate sigma factor to allow expression of the target competence genes, the same regulatory genes were induced, but the other competence genes were not. Finally, examination of the expression of 60 candidate sites not previously associated with competence identified eight additional loci that could be induced by CSP. PMID- 11029443 TI - Deletion of the min operon results in increased thermosensitivity of an ftsZ84 mutant and abnormal FtsZ ring assembly, placement, and disassembly. AB - To investigate the interaction between FtsZ and the Min system during cell division of Escherichia coli, we examined the effects of combining a well-known thermosensitive mutation of ftsZ, ftsZ84, with DeltaminCDE, a deletion of the entire min locus. Because the Min system is thought to down-regulate Z-ring assembly, the prediction was that removing minCDE might at least partially suppress the thermosensitivity of ftsZ84, which can form colonies below 42 degrees C but not at or above 42 degrees C. Contrary to expectations, the double mutant was significantly more thermosensitive than the ftsZ84 single mutant. When shifted to the new lower nonpermissive temperature, the double mutant formed long filaments mostly devoid of Z rings, suggesting a likely cause of the increased thermosensitivity. Interestingly, even at 22 degrees C, many Z rings were missing in the double mutant, and the rings that were present were predominantly at the cell poles. Of these, a large number were present only at one pole. These cells exhibited a higher than expected incidence of polar divisions, with a bias toward the newest pole. Moreover, some cells exhibited dramatically elongated septa that stained for FtsZ, suggesting that the double mutant is defective in Z-ring disassembly, and providing a possible mechanism for the polar bias. Thermoresistant suppressors of the double mutant arose that had modestly increased levels of FtsZ84. These cells also exhibited elongated septa and, in addition, produced a high frequency of branched cells. A thermoresistant suppressor of the ftsZ84 single mutant also synthesized more FtsZ84 and produced branched cells. The evidence from this study indicates that removing the Min system exposes and exacerbates the inherent defects of the FtsZ84 protein, resulting in clear septation phenotypes even at low growth temperatures. Increasing levels of FtsZ84 can suppress some, but not all, of these phenotypes. PMID- 11029444 TI - A new circadian class 2 gene, opcA, whose product is important for reductant production at night in Synechococcus elongatus PCC 7942. AB - Gene expression in the cyanobacterium Synechococcus elongatus PCC 7942 is under the control of a circadian oscillator, such that peaks and troughs of expression recur with a periodicity of about 24 h in the absence of environmental cues. This can be monitored easily as light production from luciferase gene fusions to S. elongatus promoters. All promoters seem to exhibit circadian oscillation of expression, but the phasing of peak and trough times differs among different genes. The majority of genes are designated class 1, with expression peaks near dusk or subjective dusk (the time corresponding to dusk in the absence of a diurnal cycle). A minority, of which purF is an example, have expression peaks approximately 12 h out of phase with class 1 genes. A screen of Tn5 mutants for those in which purF phasing is altered revealed a mutant that carries an insertion in the opcA gene, previously identified as essential for glucose-6 phosphate dehydrogenase function. However, a different enzymatic reporter and in vitro luciferase assays revealed that the expression pattern of the purF promoter is not altered by opcA inactivation, but rather the reduced flavin mononucleotide substrate of luciferase is limiting at the time of the natural circadian peak. The results suggest that OpcA is involved in temporally separated reductant generating pathways in S. elongatus and that it has a role outside of its function in activating glucose-6-phosphate dehydrogenase. The opcA gene, expected to be cotranscribed with fbp and zwf, was shown to have its own class 2 promoter, whereas the fbp promoter was determined to be in class 1. Thus, opcA expression is likely to be constitutive by virtue of the activity of two promoters in nearly opposite circadian phases. PMID- 11029445 TI - Mapping of the Cryptococcus neoformans MATalpha locus: presence of mating type specific mitogen-activated protein kinase cascade homologs. AB - In this study we investigated the relationship between the MATalpha locus of Cryptococcus neoformans and several MATalpha-specific mitogen-activated protein (MAP) kinase signal transduction cascade genes, including STE12alpha, STE11alpha, and STE20alpha. To resolve the location of the genes, we screened a cosmid library of the MATalpha strain B-4500 (JEC21), which was chosen for the C. neoformans genome project. We isolated several overlapping cosmids spanning a region of about 71 kb covering the entire MATalpha locus. It was found that STE12alpha, STE11alpha, and STE20alpha are imbedded within the locus rather than closely linked to the locus. Furthermore, three copies of MFalpha, the mating type alpha-pheromone gene, a MATalpha-specific myosin gene, and a pheromone receptor (CPRalpha) were identified within the locus. We created a physical map, based on the restriction enzyme BamHI, and identified both borders of the MATalpha locus. The MATalpha locus of C. neoformans is approximately 50 kb in size and is one of the largest mating type loci reported among fungi with a one locus, two-allele mating system. PMID- 11029446 TI - Vancomycin resistance is associated with serine-containing peptidoglycan in Enterococcus gallinarum. AB - In Enterococcus gallinarum SC1, a low-level vancomycin-resistant strain, only monomeric muropentapeptides with a C-terminal D-alanine were detected after growth without vancomycin. In contrast, in SC1 induced by vancomycin, as well as in AIB39, a constitutive vancomycin-resistant strain, monomeric and dimeric muropentapeptides with a C-terminal D-serine were detected. PMID- 11029447 TI - Pseudobactin biogenesis in the plant growth-promoting rhizobacterium Pseudomonas strain B10: identification and functional analysis of the L-ornithine N(5) oxygenase (psbA) gene. AB - Pseudobactin(B10), the fluorescent siderophore produced by the rhizobacterium Pseudomonas strain B10, contains the hydroxamate ligand D-N(5)-hydroxyornithine (D-N(5)-OH-Orn). We cloned the L-Orn N(5)-oxygenase (psbA) gene from a genomic library of Pseudomonas strain B10 and demonstrated that PsbA is involved in the conversion of L-Orn to its N(5)-OH derivative. PsbA shows significant similarity to microbial omega-amino acid hydroxylases containing flavin adenine dinucleotide and NADP cofactor-binding sites and the FATGY signature of the putative substrate recognition pocket. The psbA gene is monocistronic, and its transcription is negatively controlled by iron. A site-specific psbA mutant of Pseudomonas strain B10 was biochemically complemented with the precursor L-N(5)-OH-Orn, suggesting that L-Orn is hydroxylated before conversion to the D isomer. The L-Orn N(5) hydroxylase-defective mutants of Pseudomonas strain B10 and Pseudomonas aeruginosa PAO1 were much less effective than the parental strains in suppressing the growth of the phytopathogen Erwinia carotovora in iron-poor medium. The extent of in vitro inhibition of E. carotovora was strictly iron dependent and directly correlated with the amount of released siderophores. These data strengthen the role of fluorescent siderophores in biocontrol of deleterious rhizomicroorganisms. PMID- 11029448 TI - Identification and mapping of sigma-54 promoters in Chlamydia trachomatis. AB - The first sigma(54) promoters in Chlamydia trachomatis L2 were mapped upstream of hypothetical proteins CT652.1 and CT683. Comparative genomics indicated that these sigma(54) promoters and potential upstream activation binding sites are conserved in orthologous C. trachomatis D, C. trachomatis mouse pneumonitis strain, and Chlamydia pneumoniae (CWL029 and AR39) genes. PMID- 11029449 TI - Identification of the ubiD gene on the Escherichia coli chromosome. AB - The open reading frame at 86.7 min on the Escherichia coli chromosome, "yigC," complemented a ubiD mutant strain, AN66, indicating that yigC is the ubiD gene. The gene product, a 497-amino-acid-residue protein, showed extensive homology to the UPF 00096 family of proteins in the Swiss-Prot database. PMID- 11029450 TI - Polyamine transport and role of potE in response to osmotic stress in Escherichia coli. AB - When transport of polyamines in Escherichia coli was examined, putrescine excretion was observed under two different physiological conditions: (i) strictly correlated to growth and (ii) following a hyperosmotic shock. Spermidine was not excreted. Characterization of a deletion mutant showed that PotE is not involved in these transport processes. PMID- 11029451 TI - Fate of the SpoIIAB*-ADP liberated after SpoIIAB phosphorylates SpoIIAA of Bacillus subtilis. AB - Phosphorylation of SpoIIAA catalyzed by SpoIIAB helps to regulate the first sporulation-specific sigma factor, sigma(F), of Bacillus subtilis. The steady state rate of phosphorylation is known to be exceptionally slow and to be limited by the return of the protein kinase, SpoIIAB, to a catalytically active state. Previous work from this laboratory has suggested that, after catalyzing the phosphorylation, SpoIIAB is in a form (SpoIIAB*) that does not readily release ADP. We now show that the rate of release of ADP from the SpoIIAB*-ADP complex was much diminished by the presence of unreacted SpoIIAA, suggesting that SpoIIAA can form a long-lived ternary complex with SpoIIAB*-ADP in which the SpoIIAB* form is stabilized. In kinetic studies of the phosphorylation of SpoIIAA, the ternary complex SpoIIAA-SpoIIAB*-ADP could be distinguished from the short-lived complex SpoIIAA-SpoIIAB-ADP, which can be readily produced in the absence of an enzymatic reaction. PMID- 11029453 TI - Identification by gene deletion analysis of a regulator, VmsR, that controls virginiamycin biosynthesis in Streptomyces virginiae. AB - Virginiae butanolide (VB)-BarA of Streptomyces virginiae is one of the newly discovered pairs of a butyrolactone autoregulator and a corresponding receptor protein of Streptomyces species and regulates the production of the antibiotic virginiamycin (VM) in S. virginiae. The gene vmsR was found to be situated 4.7 kbp upstream of the barA gene, which encodes the VB-specific receptor. The vmsR product was predicted to be a regulator of VM biosynthesis based on its high homology to some Streptomyces pathway-specific transcriptional regulators for the biosynthetic gene clusters of polyketide antibiotics, such as Streptomyces peucetius DnrI (47.5% identity, 84. 3% similarity), which controls daunorubicin biosynthesis. A vmsR deletion mutant was created by homologous recombination. Neither virginiamycin M(1) nor virginiamycin S was produced in the vmsR mutant, while amounts of VB and BarA similar to those produced in the wild-type strain were detected. Reverse transcription-PCR analyses confirmed that the vmsR deletion had no deleterious effects on the transcription of the vmsR-surrounding genes, indicating that VmsR is a positive regulator of VM biosynthesis in S. virginiae. PMID- 11029452 TI - A second allele of eppA in Borrelia burgdorferi strain B31 is located on the previously undetected circular plasmid cp9-2. AB - Although sequence analysis of Borrelia burgdorferi isolate B31 was recently declared "complete," we found that cultures of this strain can contain a novel 9 kb circular plasmid, cp9-2. The newly described plasmid contains both sequence similarities with and differences from the previously identified B31 plasmid cp9 1 (formerly cp9). cp9-1 and cp9-2 each encode a unique allele of EppA, a putative membrane protein synthesized by B. burgdorferi during mammalian infection. PMID- 11029454 TI - Characterization of the DNA- and metal-binding properties of Vibrio anguillarum fur reveals conservation of a structural Zn(2+) ion. AB - The ferric uptake regulator, Fur, represses iron uptake and siderophore biosynthetic genes under iron-replete conditions. Here we report in vitro solution studies on Vibrio anguillarum Fur binding to the consensus 19-bp Escherichia coli iron box in the presence of several divalent metals. We found that V. anguillarum Fur binds the iron box in the presence of Mn(2+), Co(2+), Cd(2+), and to a lesser extent Ni(2+) but, unlike E. coli Fur, not in the presence of Zn(2+). We also found that V. anguillarum Fur contains a structural zinc ion that is necessary yet alone is insufficient for DNA binding. PMID- 11029455 TI - Characterization of alginate lyase from Pseudomonas syringae pv. syringae. AB - The gene encoding alginate lyase (algL) in Pseudomonas syringae pv. syringae was cloned, sequenced, and overexpressed in Escherichia coli. Alginate lyase activity was optimal when the pH was 7.0 and when assays were conducted at 42 degrees C in the presence of 0.2 M NaCl. In substrate specificity studies, AlgL from P. syringae showed a preference for deacetylated polymannuronic acid. Sequence alignment with other alginate lyases revealed conserved regions within AlgL likely to be important for the structure and/or function of the enzyme. Site directed mutagenesis of histidine and tryptophan residues at positions 204 and 207, respectively, indicated that these amino acids are critical for lyase activity. PMID- 11029456 TI - An Escherichia coli reference collection group B2- and uropathogen-associated polymorphism in the rpoS-mutS region of the E. coli chromosome. AB - Chromosomal DNAs of enterohemorrhagic, uropathogenic, and laboratory attenuated Escherichia coli strains differ in the rpoS-mutS region. Many uropathogens lack a deletion and an insertion characteristic of enterohemorrhagic strains. At the same chromosomal position, they harbor a 2.1-kb insertion of unknown origin with a base composition suggestive of horizontal gene transfer. Unlike virulence determinants associated with urinary tract infection and/or neonatal meningitis (pap or prs, sfa, kps, and hly), the 2.1-kb insertion is shared by all group B2 strains of the E. coli Reference Collection. PMID- 11029457 TI - Calcium activation of heart mitochondrial oxidative phosphorylation: rapid kinetics of mVO2, NADH, AND light scattering. AB - Parallel activation of heart mitochondria NADH and ATP production by Ca(2+) has been shown to involve the Ca(2+)-sensitive dehydrogenases and the F(0)F(1) ATPase. In the current study we hypothesize that the response time of Ca(2+) activated ATP production is rapid enough to support step changes in myocardial workload ( approximately 100 ms). To test this hypothesis, the rapid kinetics of Ca(2+) activation of mV(O(2)), [NADH], and light scattering were evaluated in isolated porcine heart mitochondria at 37 degrees C using a variety of optical techniques. The addition of Ca(2+) was associated with an initial response time (IRT) of mV(O(2)) that was dose-dependent with a minimum IRT of 0.27 +/- 0.02 s (n = 41) at 535 nm Ca(2+). The IRTs for NADH fluorescence and light scattering in response to Ca(2+) additions were similar to mV(O(2)). The Ca(2+) IRT for mV(O(2)) was significantly shorter than 1.6 mm ADP (2.36 +/- 0.47 s; p < or = 0.001, n = 13), 2.2 mm P(i) (2.32 +/- 0.29, p < or = 0.001, n = 13), or 10 mm creatine (15.6.+/-1.18 s, p < or = 0.001, n = 18) under similar experimental conditions. Calcium effects were inhibited with 8 microm ruthenium red (2.4 +/- 0.31 s; p < or = 0.001, n = 16) and reversed with EGTA (1.6 +/- 0.44; p < or = 0.01, n = 6). Estimates of Ca(2+) uptake into mitochondria using optical Ca(2+) indicators trapped in the matrix revealed a sufficiently rapid uptake to cause the metabolic effects observed. These data are consistent with the notion that extramitochondrial Ca(2+) can modify ATP production, via an increase in matrix Ca(2+) content, rapidly enough to support cardiac work transitions in vivo. PMID- 11029458 TI - Apoptosis signal-regulating kinase 1 (ASK1) is an intracellular inducer of keratinocyte differentiation. AB - Cells differentiate in response to various extracellular stimuli. This cellular response requires intracellular signaling pathways. The mitogen-activated protein (MAP) kinase cascade is a core signal transduction pathway that determines the fate of many kinds of cell. MAP kinase kinase kinase activates MAP kinase kinase, which in turn activates MAP kinase. Apoptosis signal-regulating kinase (ASK1) was identified as a MAP kinase kinase kinase involved in the stress-induced apoptosis signaling cascade that activates the SEK1-JNK and MKK3/MKK6-p38 MAP kinase cascades. Expression of the constitutively active form of ASK1 (ASK1-DeltaN) in keratinocytes induced significant morphological changes and differentiation markers, transglutaminase-1, loricrin, and involucrin. A transient increase in p21(Cip1/WAF1) reduced DNA synthesis, and cell cycle analysis verified the differentiation. p38 MAP kinase inhibitors, SB202190 and SB203580, abolished the induction of differentiation markers, transglutaminase-1, loricrin, and involucrin. In turn, the induction of differentiation with ceramide in keratinocytes caused an increase in ASK1 expression and activity. Furthermore, normal human skin expresses ASK1 protein in the upper epidermis, implicating ASK1 in in vivo keratinocyte differentiation. We propose that the ASK1-p38 MAP kinase cascade is a new intracellular regulator of keratinocyte differentiation. PMID- 11029459 TI - Oncogenic Ki-ras confers a more aggressive colon cancer phenotype through modification of transforming growth factor-beta receptor III. AB - Transforming growth factor-beta1 (TGF-beta1) can act as a tumor suppressor or a tumor promoter depending on the characteristics of the malignant cell. Each of three Ki-ras(G12V) transfectants of HD6-4 colon cancer cells had been shown to be more aggressive in vivo than controls in earlier studies (Yan, Z., Chen, M., Perucho, M., and Friedman, E. (1997) J. Biol. Chem. 272, 30928-30936). We now show that stable expression of oncogenic Ki-ras(G12V) converts the HD6-4 colon cancer cell line from insensitive to TGF-beta1 to growth-promoted by TGF-beta1. Each of three Ki-ras(G12V) transfectants responded to TGF-beta1 by an increase in proliferation and by decreasing the abundance of the Cdk inhibitor p21 and the tumor suppressor PTEN, whereas each of three wild-type Ki-ras transfectants remained unresponsive to TGF-beta1. The wild-type Ki-ras transfectants lack functional TGF-beta receptors, whereas all three Ki-ras(G12V) transfectants expressed functional TGF-beta receptors that bound (125)I-TGF-beta1. The previous studies showed that in cells with wild-type Ki-ras, TGF-beta receptors were not mutated, and receptor proteins were transported to the cell surface, but post translational modification of TGF-beta receptor III (TbetaRIII) was incomplete. We now show that the betaglycan form of TbetaRIII is highly modified following translation when transiently expressed in Ki-ras(G12V) cells, whereas no such post-translational modification of TbetaRIII occurs in control cells. Antisense oligonucleotides directed to Ki-Ras decreased both TbetaRIII post-translational modification in Ki-ras(G12V) cells and TGF-beta1 down-regulation of p21, demonstrating the direct effect of mutant Ras. Therefore, one mechanism by which mutant Ki-Ras confers a more aggressive tumor phenotype is by enhancing TbetaRIII post-translational modification. PMID- 11029460 TI - Fe3+ coordination and redox properties of a bacterial transferrin. AB - The Fe(3+) binding site of recombinant nFbp, a ferric-binding protein found in the periplasmic space of pathogenic Neisseria, has been characterized by physicochemical techniques. An effective Fe(3+) binding constant in the presence of 350 microm phosphate at pH 6.5 and 25 degrees C was determined as 2.4 x 10(18) m(-1). EPR spectra for the recombinant Fe(3+)nFbp gave g' = 4.3 and 9 signals characteristic of high spin Fe(3+) in a strong ligand field of low (orthorhombic) symmetry. (31)P NMR experiments demonstrated the presence of bound phosphate in the holo form of nFbp and showed that phosphate can be dialyzed away in the absence of Fe(3+) in apo-nFbp. Finally, an uncorrected Fe(3+/2+) redox potential for Fe-nFbp was determined to be -290 mV (NHE) at pH 6.5, 20 degrees C. Whereas our findings show that nFbp and mammalian transferrin have similar Fe(3+) binding constants and EPR spectra, they differ greatly in their redox potentials. This has implications for the mechanism of Fe transport across the periplasmic space of Gram-negative bacteria. PMID- 11029461 TI - Phosphatidylinositol 4,5-bisphosphate is acting as a signal molecule in alpha(1) adrenergic pathway via the modulation of acetylcholine-activated K(+) channels in mouse atrial myocytes. AB - We have investigated the effect of alpha(1)-adrenergic agonist phenylephrine (PE) on acetylcholine-activated K(+) currents (I(KACh)). I(KACh) was recorded in mouse atrial myocytes using the patch clamp technique. I(KACh) was activated by 10 microm ACh and the current decreased by 44.27 +/- 2.38% (n = 12) during 4 min due to ACh-induced desensitization. When PE was applied with ACh, the extent of desensitization was markedly increased to 69.34 +/- 2.22% (n = 9), indicating the presence of PE-induced desensitization. I(KACh) was fully recovered from desensitization after a 6-min washout. PE-induced desensitization of I(KACh) was not affected by protein kinase C inhibitor, calphostin C, but abolished by phospholipase C (PLC) inhibitor, neomycin. When phophatidylinositol 4,5 bisphosphate (PIP(2)) replenishment was blocked by wortmannin (an inhibitor of phophatidylinositol 3-kinase and phophatidylinositol 4-kinase), desensitization of I(KACh) in the presence of PE was further increased (97.25 +/- 7.63%, n = 6). Furthermore, the recovery from PE-induced desensitization was inhibited, and the amplitude of I(KACh) at the second exposure after washout was reduced to 19.65 +/ 2.61% (n = 6) of the preceding level. These data suggest that the K(ACh) channel is modulated by PE through PLC stimulation and depletion of PIP(2). PMID- 11029462 TI - Molecular cloning and characterization of a human beta-Gal-3'-sulfotransferase that acts on both type 1 and type 2 (Gal beta 1-3/1-4GlcNAc-R) oligosaccharides. AB - A novel sulfotransferase gene (designated GP3ST) was identified on human chromosome 2q37.3 based on its similarity to the cerebroside 3'-sulfotransferase (CST) cDNA (Honke, K., Tsuda, M., Hirahara, Y., Ishii, A., Makita, A., and Wada, Y. (1997) J. Biol. Chem. 272, 4864-4868). A full-length cDNA was obtained by reverse transcription-polymerase chain reaction and 5'- and 3'-rapid amplification of cDNA ends analyses of human colon mRNA. The isolated cDNA clone predicts that the protein is a type II transmembrane protein composed of 398 amino acid residues. The amino acid sequence indicates 33% identity to the human CST sequence. A recombinant protein that is expressed in COS-1 cells showed no CST activity, but did show sulfotransferase activities toward oligosaccharides containing nonreducing beta-galactosides such as N-acetyllactosamine, lactose, lacto-N-tetraose (Lc4), lacto-N-neotetraose (nLc4), and Gal beta 1-3GalNAc alpha benzyl (O-glycan core 1 oligosaccharide). To characterize the cloned sulfotransferase, a sulfotransferase assay method was developed that uses pyridylaminated (PA) Lc4 and nLc4 as enzyme substrates. The enzyme product using PA-Lc4 as an acceptor was identified as HSO(3)-3Gal beta 1-3GlcNAc beta 1-3Gal beta 1- 4Glc-PA by two-dimensional (1)H NMR. Kinetics studies suggested that GP3ST is able to act on both type 1 (Gal beta 1-3GlcNAc-R) and type 2 (Gal beta 1 4GlcNAc-R) chains with a similar efficiency. In situ hybridization demonstrated that the GP3ST gene is expressed in epithelial cells lining the lower to middle layer of the crypts in colonic mucosa, hepatocytes surrounding the central vein of the liver, extravillous cytotrophoblasts in the basal plate and septum of the placenta, renal tubules of the kidney, and neuronal cells of the cerebral cortex. The results of this study indicate the existence of a novel beta-Gal-3' sulfotransferase gene family. PMID- 11029463 TI - Selective resistance to parathyroid hormone caused by a novel uncoupling mutation in the carboxyl terminus of G alpha(s). A cause of pseudohypoparathyroidism type Ib. AB - G(s) is a heterotrimeric (alpha, beta, and gamma chains) G protein that couples heptahelical plasma membrane receptors to stimulation of adenylyl cyclase. Inactivation of one GNAS1 gene allele encoding the alpha chain of G(s) (G alpha(s)) causes pseudohypoparathyroidism type Ia. Affected subjects have resistance to parathyroid hormone (PTH) and other hormones that activate adenylyl cyclase plus somatic features termed Albright hereditary osteodystrophy. By contrast, subjects with pseudohypoparathyroidism type Ib have hormone resistance that is limited to PTH and lack Albright hereditary osteodystrophy. The molecular basis for pseudohypoparathyroidism type Ib is unknown. We analyzed the GNAS1 gene for mutations using polymerase chain reaction to amplify genomic DNA from three brothers with pseudohypoparathyroidism type Ib. We identified a novel heterozygous 3-base pair deletion causing loss of isoleucine 382 in the three affected boys and their clinically unaffected mother and maternal grandfather. This mutation was absent in other family members and 15 additional unrelated subjects with pseudohypoparathyroidism type Ib. To characterize the signaling properties of the mutant G alpha(s), we used site-directed mutagenesis to introduce the isoleucine 382 deletion into a wild type G alpha(s) cDNA, transfected HEK293 cells with either wild type or mutant G alpha(s) cDNA, plus cDNAs encoding heptahelical receptors for PTH, thyrotropic hormone, or luteinizing hormone, and we measured cAMP production in response to hormone stimulation. The mutant G alpha(s) protein was unable to interact with the receptor for PTH but showed normal coupling to the other coexpressed heptahelical receptors. These results provide evidence of selective uncoupling of the mutant G alpha(s) from PTH receptors and explain PTH-specific hormone resistance in these three brothers with pseudohypoparathyroidism type Ib. The absence of PTH resistance in the mother and maternal grandfather who carry the same mutation is consistent with current models of paternal imprinting of the GNAS1 gene. PMID- 11029464 TI - Alteration in calcium handling at the subcellular level in mdx myotubes. AB - In this study, we have tested the hypothesis that augmented [Ca(2+)] in subcellular regions or organelles, which are known to play a key role in cell survival, is the missing link between Ca(2+) homeostasis alterations and muscular degeneration associated with muscular dystrophy. To this end, different targeted chimeras of the Ca(2+)-sensitive photoprotein aequorin have been transiently expressed in subcellular compartments of skeletal myotubes of mdx mice, the animal model of Duchenne muscular dystrophy. Direct measurements of the [Ca(2+)] in the sarcoplasmic reticulum, [Ca(2+)](sr), show a higher steady state level at rest and a larger drop after KCl-induced depolarization in mdx compared with control myotubes. The peaks in [Ca(2+)] occurring in the mitochondrial matrix of mdx myotubes are significantly larger than in controls upon KCl-induced depolarization or caffeine application. The augmented response of mitochondria precedes the alterations in the Ca(2+) responses of the cytosol and of the cytoplasmic region beneath the membrane, which become significant only at a later stage of myotube differentiation. Taking into account the key role played by mitochondria Ca(2+) handling in the control of cell death, our data suggest that mitochondria are potential targets of impaired Ca(2+) homeostasis in muscular dystrophy. PMID- 11029465 TI - Activation of the Arp2/3 complex by the Listeria acta protein. Acta binds two actin monomers and three subunits of the Arp2/3 complex. AB - ActA is a bacterially encoded protein that enables Listeria monocytogenes to hijack the host cell actin cytoskeleton. It promotes Arp2/3-dependent actin nucleation, but its interactions with cellular components of the nucleation machinery are not well understood. Here we show that two domains of ActA (residues 85-104 and 121-138) with sequence similarity to WASP homology 2 domains bind two actin monomers with submicromolar affinity. ActA binds Arp2/3 with a K(d) of 0.6 microm and competes for binding with the WASP family proteins N-WASP and Scar1. By chemical cross-linking, ActA, N-WASP, and Scar1 contact the same three subunits of the Arp2/3 complex, p40, Arp2, and Arp3. Interestingly, profilin competes with ActA for binding of Arp2/3, but actophorin (cofilin) does not. The minimal Arp2/3-binding site of ActA (residues 144-170) is C-terminal to both actin-binding sites and shares sequence homology with Arp2/3-binding regions of WASP family proteins. The maximal activity at saturating concentrations of ActA is identical to the most active domains of the WASP family proteins. We propose that ActA and endogenous WASP family proteins promote Arp2/3-dependent nucleation by similar mechanisms and require simultaneous binding of Arp2 and Arp3. PMID- 11029466 TI - Arabidopsis eIF3e (INT-6) associates with both eIF3c and the COP9 signalosome subunit CSN7. AB - The Arabidopsis COP9 signalosome is a multisubunit repressor of photomorphogenesis that is conserved among eukaryotes. This complex may have a general role in development. As a step in dissecting the biochemical mode of action of the COP9 signalosome, we determined the sequence of proteins that copurify with this complex. Here we describe the association between components of the COP9 signalosome (CSN1, CSN7, and CSN8) and two subunits of eukaryotic translation initiation factor 3 (eIF3), eIF3e (p48, known also as INT-6) and eIF3c (p105). To obtain a biochemical marker for Arabidopsis eIF3, we cloned the Arabidopsis ortholog of the eIF3 subunit eIF3b (PRT1). eIF3e coimmunoprecipitated with CSN7, and eIF3c coimmunoprecipitated with eIF3e, eIF3b, CSN8, and CSN1. eIF3e directly interacted with CSN7 and eIF3c. However, eIF3e and eIF3b cofractionated by gel filtration chromatography in a complex that was larger than the COP9 signalosome. Whereas eIF3, as detected through eIF3b, localized solely to the cytoplasm, eIF3e, like CSN7, was also found in the nucleus. This suggests that eIF3e and eIF3c are probably components of multiple complexes and that eIF3e and eIF3c associate with subunits of the COP9 signalosome, even though they are not components of the COP9 signalosome core complex. This interaction may allow for translational control by the COP9 signalosome. PMID- 11029467 TI - Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide inhibit nuclear factor-kappa B-dependent gene activation at multiple levels in the human monocytic cell line THP-1. AB - The neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) suppress monocyte/macrophage production of proinflammatory agents. The transcription factor NF-kappa B regulates the transcription of most agents. VIP/PACAP inhibit NF-kappa B transactivation in the lipopolysaccharide-stimulated human monocytic cell line THP-1 at multiple levels. First, VIP/PACAP inhibit p65 nuclear translocation and NF-kappa B DNA binding by stabilizing the inhibitor I kappa B alpha. Second, VIP/PACAP induce phosphorylation of the CRE-binding protein (CREB) and its binding to the CREB binding protein (CBP). This results in a decrease in p65.CBP complexes, which further reduces NF-kappa B transactivation. Third, VIP and PACAP reduce the phosphorylation of the TATA box-binding protein (TBP), resulting in a reduction in TBP binding to both p65 and the TATA box. All these effects are mediated through the specific receptor VPAC1. The cAMP/cAMP-dependent protein kinase pathway mediates the effects on CBP and TBP, whereas a cAMP-independent pathway is the major transducer for the effects on p65 nuclear translocation. Since NF kappaB represents a focal point for various stimuli and induces the expression of many proinflammatory genes, its targeting by VIP and PACAP positions them as important anti-inflammatory agents. The VIP/PACAP inhibition of NF-kappa B at various levels and through different transduction pathways could offer a significant advantage over other anti-inflammatory agents. PMID- 11029468 TI - Kinetic basis for activation of CDK2/cyclin A by phosphorylation. AB - The activation of most protein kinases requires phosphorylation at a conserved site within a structurally defined segment termed the activation loop. A classic example is the regulation of the cell cycle control enzyme, CDK2/cyclin A, in which catalytic activation depends on phosphorylation at Thr(160) in CDK2. The structural consequences of phosphorylation have been revealed by x-ray crystallographic studies on CDK2/cyclin A and include changes in conformation, mainly of the activation loop. Here, we describe the kinetic basis for activation by phosphorylation in CDK2/cyclin A. Phosphorylation results in a 100,000-fold increase in catalytic efficiency and an approximate 1,000-fold increase in the overall turnover rate. The effects of phosphorylation on the individual steps in the catalytic reaction pathway were determined using solvent viscosometric techniques. It was found that the increase in catalytic power arises mainly from a 3,000-fold increase in the rate of the phosphoryl group transfer step with a more moderate increase in substrate binding affinity. In contrast, the rate of phosphoryl group transfer in the ATPase pathway was unaffected by phosphorylation, demonstrating that phosphorylation at Thr(160) does not serve to stabilize ATP in the ATPase reaction. Thus, we hypothesize that the role of phosphorylation in the kinase reaction may be to specifically stabilize the peptide phosphoacceptor group. PMID- 11029469 TI - Selective targeting of MAPKs to the ETS domain transcription factor SAP-1. AB - MAPK pathways play important roles in regulating the key cellular processes of proliferation, differentiation, and apoptosis. There are multiple MAPK pathways, which are subject to different regulatory cues. It is important that these pathways maintain specificity in signaling to elicit the activation of a specific program of gene expression. MAPK-docking domains in several transcription factors have been shown to play important roles in determining the specificity and efficiency of their phosphorylation by MAPKs. Here we investigate the mechanisms by which MAPKs are targeted to the ETS domain transcription factor SAP-1. We demonstrate that SAP-1 contains two different domains that are required for its efficient phosphorylation in vitro and activation in vivo by ERK2 and a subset of p38 MAPKs. The D-domain is closely related to other MAPK-docking domains, but exhibits a novel specificity and serves to promote selective targeting of ERK2, p38alpha, and p38beta(2) to SAP-1. A second important region, the FXF motif, also plays an important role in directing MAPKs to phosphorylate SAP-1. The FXF motif promotes targeting by ERK2 and, to a lesser extent, p38alpha, but not p38beta(2). Our data therefore demonstrate that a modular system of motifs is responsible for directing specific MAPK subtypes to SAP-1, but also point to important distinctions in the mechanism of action of the D-domain and FXF motif. PMID- 11029470 TI - Differential regulation of growth and checkpoint control mediated by a Cdc25 mitotic phosphatase from Pneumocystis carinii. AB - Pneumocystis carinii is an opportunistic fungal pathogen phylogenetically related to the fission yeast Schizosaccharomyces pombe. P. carinii causes severe pneumonia in immunocompromised patients with AIDS and malignancies. Although the life cycle of P. carinii remains poorly characterized, morphologic studies of infected lung tissue indicate that P. carinii alternates between numerous small trophic forms and fewer large cystic forms. To understand further the molecular mechanisms that regulate progression of the cell cycle of P. carinii, we have sought to identify and characterize genes in P. carinii that are important regulators of eukaryotic cell cycle progression. In this study, we have isolated a cDNA from P. carinii that exhibits significant homology, but unique functional characteristics, to the mitotic phosphatase Cdc25 found in S. pombe. P. carinii Cdc25 was shown to rescue growth of the temperature-sensitive S. pombe cdc25-22 strain and thus provides an additional tool to investigate the unique P. carinii life cycle. Although P. carinii Cdc25 could also restore the DNA damage checkpoint in cdc25-22 cells, it was unable to restore fully the DNA replication checkpoint. The dissociation of checkpoint control at the level of Cdc25 indicates that Cdc25 may be under distinct regulatory control in mediating checkpoint signaling. PMID- 11029471 TI - c-Jun N-terminal kinase activation in Xenopus laevis eggs and embryos. A possible non-genomic role for the JNK signaling pathway. AB - The c-Jun N-terminal kinases (JNKs) are members of the mitogen-activated protein kinase family that play critical roles in stress responses and apoptosis. We have discovered that JNK is present in Xenopus oocytes, an experimental system that offers a variety of powerful experimental approaches to questions of protein function and regulation. Like ERK2/p42 MAPK, JNK is activated just prior to germinal vesicle breakdown during Xenopus oocyte maturation and remains active throughout meiosis I and II. However, unlike p42 MAPK, which is inactivated about 30 min after eggs are fertilized or parthenogenetically activated, JNK stays constitutively active until the early gastrula stage of embryogenesis. These findings suggest that the JNK pathway may play a role in oocyte maturation and embryogenesis. JNK was activated by microinjection of Mos, by activation of an estrogen-inducible form of Raf, and by a constitutively active MEK-1 (MEK R4F), indicating that the p42 MAPK cascade can trigger JNK activation. However, the MEK inhibitor U0126 blocked progesterone-induced p42 MAPK activation but not progesterone-induced JNK activation. Thus, progesterone can stimulate JNK activation both through the MEK/p42 MAPK pathway and through MEK/p42 MAPK independent pathways. Many of the key substrates of JNKs identified to date are transcriptional regulators. However, since transcription is not required for germinal vesicle breakdown in progesterone-treated oocytes or for the early embryonic cell cycles, our findings suggest that in these contexts the JNK pathway exerts nongenomic effects. PMID- 11029472 TI - Roles of ionic residues of the C1 domain in protein kinase C-alpha activation and the origin of phosphatidylserine specificity. AB - On the basis of extensive structure-function studies of protein kinase C-alpha (PKC-alpha), we have proposed an activation mechanism for conventional PKCs in which the C2 domain and the C1 domain interact sequentially with membranes (Medkova, M., and Cho, W. (1999) J. Biol. Chem. 274, 19852-19861). To further elucidate the interactions between the C1 and C2 domains during PKC activation and the origin of phosphatidylserine specificity, we mutated several charged residues in two C1 domains (C1a and C1b) of PKC-alpha. We then measured the membrane binding affinities, activities, and monolayer penetration of these mutants. Results indicate that cationic residues of the C1a domain, most notably Arg(77), interact nonspecifically with anionic phospholipids prior to the membrane penetration of hydrophobic residues. The mutation of a single aspartate (Asp(55)) in the C1a domain to Ala or Lys resulted in dramatically reduced phosphatidylserine specificity in vesicle binding, activity, and monolayer penetration. In particular, D55A showed much higher vesicle affinity, activity, and monolayer penetration power than wild type under nonactivating conditions, i.e. with phosphatidylglycerol and in the absence of Ca(2+), indicating that Asp(55) is involved in the tethering of the C1a domain to another part of PKC alpha, which keeps it in an inactive conformation at the resting state. Based on these results, we propose a refined model for the activation of conventional PKC, in which phosphatidylserine specifically disrupts the C1a domain tethering by competing with Asp(55), which then leads to membrane penetration and diacylglycerol binding of the C1a domain and PKC activation. PMID- 11029473 TI - Diabetic nephropathy--where next? PMID- 11029474 TI - Snoring, sleep apnoea and stroke: chicken or scrambled egg? PMID- 11029475 TI - Neurological deficits in solvent-exposed painters: a syndrome including impaired colour vision, cognitive defects, tremor and loss of vibration sensation. AB - Five individuals are described who had participated in a study of former dockyard painters. All had worked between 16 years and 45 years as industrial painters, much of the time inside ships. All underwent structured neurological examination, colour vision testing (allowing calculation of a colour confusion index corrected for age and alcohol), and detailed psychometric testing. An occupational history sufficient to allow estimation of past exposure to solvents was taken. All gave a history of exposure to high concentrations of solvents at work, and several described episodes of acute narcosis. All showed neurological deficits and some had overt neurological disease, although in no case had this previously been linked to their work. The most striking features, sufficient to constitute a syndrome, were acquired blue-yellow colour vision deficits, coarse tremor, impaired vibration sensation in the legs and cognitive impairment. Their estimated cumulative exposures to solvents ranged between the equivalent of 13 and 37 calendar years working at the Occupational Exposure Standard concentration (OES years). This study for the first time gives an indication of the concentrations of solvents likely to lead to serious neurological disease in humans. It serves as a reminder to physicians to take an occupational history from patients with obscure neurological or psychological impairment. PMID- 11029476 TI - UK patients with deep-vein thrombosis can be safely treated as out-patients. AB - Deep-vein thrombosis (DVT) affects approximately 1:1000 people, approximately 750 cases/year in a Health Authority of average size. Prior to 1992, patients presenting with DVT were usually admitted for treatment with unfractionated heparin (UFH) over a 5-day period, but pressures on medical admissions have prompted many hospitals to review conditions which could be managed at home. Three different pilot studies commenced in 1996 at three centres in the UK. After 6 months, the protocols used were integrated into the normal care plan of the hospital. In total, 5191 patients were assessed, of whom 1347 were either venogram or Doppler ultrasound positive. Overall 1138 (82%) were treated as out patients, 75% presenting during 'working hours'. Only 12 patients were readmitted, one with a clinically significant PE. Success was attributed to three factors: assignment of a key person as the project co-ordinator; referral of patients directly to permanent, dedicated staff, either on the Medical Admissions Unit (MAU) or the Accident and Emergency (A&E) department; and the introduction of dedicated anticoagulation nurses. In the 6-month period following initial therapy, complications were well below those in previously published studies. Most patients with DVT in the UK can be treated safely and effectively without being admitted to hospital. PMID- 11029477 TI - Use of large medical databases to study associations between diseases. AB - We describe the use of a dataset of statistical medical records, the Oxford Record Linkage Study (ORLS), to identify diseases which occur together more commonly (association), or less commonly (dissociation), than their individual frequencies in the population would predict. We investigated some conditions known or suspected to enhance the subsequent risk of cancer, some conditions thought to be linked with schizophrenia, and some associations between conditions with a known autoimmune component. Diseases may occur in combination more often (or less often) than expected by chance because one predisposes to (or protects against) another or because they share environmental and/or genetic mechanisms in common. The investigation of such associations can yield important information for clinicians interested in potential disease sequelae, for epidemiologists trying to understand disease aetiology, and for geneticists attempting to determine the genetic basis of variation in disease course among individuals. We suggest that, through the use of datasets like the ORLS, it will be possible to 'map' comprehensively the phenomic expression of co-occurring diseases. PMID- 11029478 TI - Prevalence of hip fracture risk factors in women aged 70 years and over. AB - We estimated the prevalence of common risk factors for hip fracture and the numbers needed to treat (NNT) to prevent a hip fracture in various high-risk population groups, using a postal risk factor survey of women aged 70 years and above from General Practices in Grampian and Yorkshire. Recorded risk factors included prior fracture of any type; low body weight; smoking; and family history of fracture. The prevalence rates of hip fracture risk factors were 34%, 7% and 11% for previous fracture, maternal hip fracture and smoking, respectively for the Grampian practices (low body weight being defined as falling in the lowest quartile) and 34%, 7% and 7% for a single practice in the York area. Applying previously published estimates of risk, NNT analysis produced a value of about 300 for women with no risk factors, whilst for women with three risk factors it was between 32 and 71, depending on which risk factors were present and assuming intervention reduced fracture rates by 30% or 50%. Groups of women at high risk of hip fracture can easily be identified in primary care and offered treatment, with realistic prospects of hip fracture prevention. PMID- 11029479 TI - Familial Mediterranean fever and systemic amyloidosis in untreated Turkish patients. AB - We compared the frequencies of seven MEFV mutations (M694V, M680I, V726A, M694I, K695R, R761H, E148Q) and the clinical findings in 20 Turkish FMF patients who had not developed amyloidosis by the age of 40 years in the absence of colchicine therapy, with those in 27 Turkish amyloidosis patients. No mutation frequency, including that of M694V, was different between the two groups. Family history of amyloidosis and parental consanguinity were noted to be higher in the amyloidosis group. The seven mutations do not appear to be sufficient to explain the development of amyloidosis in Turkish FMF patients. Other genetic factors may be important for this association. PMID- 11029480 TI - The patient with a systolic murmur: severe aortic stenosis may be missed during cardiovascular examination. AB - Significant aortic stenosis is prevalent amongst elderly people. It may be subclinical, manifesting only as a murmur, but can still cause unexpected death with little warning after symptoms develop. Recent studies have highlighted the unreliability of the classical clinical signs of severe aortic stenosis, leading to concern that some patients may not be referred appropriately for echocardiography. Here, we review the evidence for the accuracy of each sign. We suggest that the assessment of the patient with a systolic murmur should be reappraised, and offer guidelines toward improving the recognition of aortic stenosis in the community. PMID- 11029481 TI - Testosterone: a natural tonic for the failing heart? PMID- 11029482 TI - A.M. Cooke: an informal memoir. PMID- 11029483 TI - Oestrogen and women's heart disease: ESPRIT-UK. PMID- 11029484 TI - Controlling hypertension in the elderly. PMID- 11029485 TI - Silent myocardial ischaemia after coronary artery bypass graft. PMID- 11029486 TI - Blood supply, oxygenation status and metabolic micromilieu of breast cancers: characterization and therapeutic relevance. AB - The metabolic microenvironment of a tumor is predominantly determined by the efficacy of blood flow, flux parameters (such as diffusion and convective currents in the interstitial space) and metabolic rates. The most important factors in this context include oxygen and nutrient supply, tissue pH and the bioenergetic status. It is now widely accepted that the metabolic microenvironment of a tumor can dramatically influence a range of factors such as proliferation rate, cell cycle position, growth rate and the development of apoptosis and necrosis. At the same time, these parameters can have an impact on tumor detection, therapeutic response to conventional irradiation, some chemotherapy agents and other non-surgical treatment modalities, while also having the capacity to modulate malignant progression (e.g., enhanced local spread, metastasis). The metabolic microenvironment appears to have a potentially important role to play in the prediction of long-term treatment outcome, and thus might be useful as a prognostic factor. Currently available information related to the parameters defining the metabolic microenvironment in breast cancers are presented in this review. According to these data, significant variations in these relevant factors are likely to occur between different locations within a tumor, and between tumors of the same clinical size, stage, grade and histology. The to treatment are also considered. PMID- 11029487 TI - Forskolin increases the expression of the pancreatic tumor antigen, Nd2, and uptake of Nd2 antibody. AB - Nd2 antibody recognizes an antigen that is tumor specific in pancreas. It has been used successfully in clinical radioimmunodetection studies to identify exocrine pancreatic tumors. In the present study we show that the uptake of radiolabeled Nd2 antibody by SW1990 pancreatic carcinoma cells was increased by the adenyl cyclase activator, forskolin. Dibutyryl cyclic AMP and forskolin were both effective in increasing the level of Nd2 antigen in SW1990 cells. Immunoprecipitation studies showed that the Nd2 epitope is associated with MUC1 mucin. Forskolin increased Nd2/MUC1 antigen in both a membrane fraction and a high buoyant density mucin-like fraction. Nd2 immunoreactivity was reduced by treatment of mucins with proteases and beta-mercaptoethanol. Immunohistochemical studies showed that periodate catalyzed beta-elimination greatly reduced Nd2 immunoreactivity. These results suggest that the Nd2 epitope is unusual in having characteristics of both a peptide and a carbohydrate, protease and conformation sensitivities and involvement of O-linked oligosaccharides. Nd2 antibody does not react with several known pancreatic cancer antigens. In summary, activation of the cyclic AMP pathway increased cellular uptake of Nd2 antibody and the cellular expression of the tumor-specific, mucin-associated Nd2 antigen. These results suggest a means of improving the effectiveness of monoclonal antibodies in targeting tumor antigens for the diagnosis and treatment of malignancy. PMID- 11029488 TI - Dihydropyrimidine dehydrogenase activity and mRNA expression in advanced gastric cancer analyzed in relation to effectiveness of preoperative 5-fluorouracil-based chemotherapy. AB - Dihydroxypyrimidine dehydrogenase (DPD) is an enzyme involved in degradation and inactivation of 5-fluorouracil (5-FU). The amount of its expression in a tumor is thought to be a factor determining the response of the tumor to 5-FU therapy. We compared DPD activity and DPD mRNA expression in resected tumors between two groups of patients, i.e., a group of 14 patients with advanced gastric cancer who received preoperative chemotherapy (neoadjuvant chemotherapy; NAC) and surgery and a group of 24 patients with advanced gastric cancer who underwent surgery without preoperative chemotherapy. Tumor DPD activity was found to correlate well with tumor DPD mRNA expression. In the surgery alone group, DPD activity decreased significantly as the tumor stage advanced. This change was not observed in the NAC plus surgery group. Neither tumor depth (T factor) nor lymph node metastasis was found to correlate with DPD activity. Patients who responded to preoperative chemotherapy had lower DPD mRNA levels. Based on these results, we anticipate that measurement of DPD expression in clinical specimens may be clinically useful in managing advanced gastric cancer. PMID- 11029490 TI - Genetic alterations in metastatic renal cell carcinoma detected by comparative genomic hybridization: correlation with clinical and histological data. AB - In order to optimize the management of patients with renal cell carcinoma (RCC) it is important to define the genetic risk for metastatic disease. In this study we performed comparative genomic hybridization (CGH) on metastatic tumors aiming at the identification of genetic alterations associated with metastatic disease. We analyzed 46 renal tumors along with their metastases, and 15 non-metastatic renal tumors. Tumors were classified pathologically according to the Heidelberg classification of RCC, and staged according to the TNM-system. Standard CGH was performed using microdissected archival tissues and DOP-PCR. The average numbers of chromosomal aberrations per tumor were 3.0, 2.1 and 3.9 in patients without metastasis, in patients who developed metastases after a two-year latency period (late onset of metastatic disease) and in patients who developed metastases within two years after therapy of the primary tumor (early onset of metastatic disease). CGH revealed chromosomal aberrations in 91% of primary metastatic tumors. Deletions or losses of chromosomes 9 (26% vs 6%), 10 (21% vs 6%) and 18 (23% vs 0) and 17 (28% vs 7%) occurred more often in metastatic tumors than in non-metastatic tumors. Furthermore, these aberrations were more common in patients with early metastases. CGH analysis of 40 pairs of primary RCCs and their corresponding metastasis revealed similar aberrations in 70% of cases. In 30%, however, metastases showed additional chromosomal aberrations not detected in the corresponding primary tumors. In conclusion, we identified genetic alterations associated with metastatic disease in RCC which could be useful for predicting prognosis. Genetic changes leading to metastases occurred early in tumorigenesis of metastatic tumors. PMID- 11029489 TI - The Rb family of cell cycle regulatory factors: clinical implications. AB - The retinoblastoma gene family is composed of three members: the product of the retinoblastoma gene (pRb), which is one of the most well studied tumor suppressor genes and two related proteins, pRb2/p130 and p107, which have been shown to be structurally and functionally similar to pRb. The three retinoblastoma family members show growth suppressive properties, although the growth arrest mediated by each of the three pocket regions of the proteins is not identical. This supports the idea that although the three members may complement each other, they are not fully functional or redundant. Among the three family members, the retinoblastoma-related gene product pRb2/p130 is a tumor suppressor gene and an effective candidate target for gene therapy approach. The aim of this review is to examine the role of the Rb family members in growth regulation discussing their putative prognostic and therapeutical impact in human cancer. PMID- 11029491 TI - Expression in hepatomas and chromosomal localization of rat protein phosphatase 5 gene. AB - The serine/threonine protein phosphatase type 5 (PP5) gene expression was ubiquitously observed among all the tissues examined, but being at the lowest level in the liver. The levels of PP5 mRNA were markedly elevated in rat highly malignant ascites hepatomas, while those in several rat primary hepatomas were slightly increased and those in the regenerating livers were not elevated at all compared to the control liver. The PP5 gene was mapped to rat chromosome 1q22.1. This region has been identified to have linkage homology among human, mouse and rat, and is known to be associated with several tumor types. Taken together, the present results strongly suggest important roles of PP5 in tumorigenesis. PMID- 11029492 TI - Telomerase activity and expression of apoptosis and anti-apoptosis regulators in the progression pathway of human melanoma. AB - The stages defining the progression pathway of human melanoma are atypical nevi, the precursor lesions and risk markers of melanoma, melanoma in situ and melanoma in the radical growth phase (RGP), which represent the early stages of melanoma development, and primary melanoma in the vertical growth phase (VGP) and melanoma in the metastatic growth phase (MGP), which are the advanced stages of the disease. Unlike cells obtained from VGP and MGP melanomas, which can be established as cell lines, cells derived from atypical nevi, melanoma in situ, and RGP melanoma cannot be propagated in vitro. Thus, information regarding molecular markers that may be differentially expressed in the early versus the advanced stages of this disease can only be obtained from the analysis of specimens. Since activation of telomerase and deregulation of apoptosis contribute to the pathogenesis of a significant number of human malignancies, we conducted a study, using nevus and melanoma specimens, to determine at what stage in the progression pathway of melanoma, telomerase activity can first be detected, and whether concordant with telomerase activation, one might observe a stage-specific switch from expression of promoters to inhibitors of apoptosis. The findings described here, demonstrate telomerase activity in some but not all MGP melanomas and not in any of the preceding pathological stages, and no apparent imbalance between pro- and anti-apoptotic markers in telomerase-positive MGP melanomas compared to telomerase-negative nevi and telomerase-negative VGP and MGP melanomas. PMID- 11029493 TI - p53 accumulation confers prognostic information in resectable adenocarcinomas with ductal but not with intestinal differentiation in the pancreatic head. AB - The aim of the study was to examine the relation between p53 protein accumulation, clinicopathological variables and prognosis in resectable adenocarcinomas of the pancreatic head. The clinical records and tissue specimens of 82 consecutive patients resected for adenocarcinomas located in the head of the pancreas were reviewed retrospectively. Formalin-fixed and paraffin-embedded specimens from each tumour were stained with the monoclonal antibody DO7, and the nuclear p53 positivity within each tumour was assessed. Histopathological reclassification showed that 60 tumours exhibited ductal differentiation and 22 tumours intestinal differentiation. Twenty-five percent (15/60) of the ductal tumours and 50% (11/22) of the intestinal tumours were positive for p53 accumulation. p53 immunoreactivity was significantly correlated to a worse prognosis in the tumours of ductal differentiation, with median survival 0.76 years for p53 positive and 1.44 years for p53 negative patients. The p53 positivity of tumours with intestinal differentiation showed no such correlation. No correlation was found between p53 accumulation and other known prognostic factors in either the ductal or the intestinal type of tumours. Our results indicate that the tumour biology of ductal adenocarcinomas differs significantly from that of adenocarcinomas of the intestinal type located in the pancreatic head, and that p53 accumulation confers a worse prognosis only of ductal tumours. Subclassification of these tumours based on type of differentiation is therefore suggested since periampullary tumours include ductally as well as intestinally differentiated adenocarcinomas. PMID- 11029494 TI - Expression of hyaluronan receptors CD44 and RHAMM in stomach cancers: relevance with tumor progression. AB - Interactions of hyaluronic acid (HA) with its binding proteins CD44 and RHAMM (receptor for HA-mediating motility) have been proposed to be important in promoting tumor progression and dissemination. However, a comparative study of their expression patterns in stomach cancer and its associated lesions is not yet available. To address this issue, the combined examinations of pathology, immunocytochemistry and Western blot hybridization were performed on advanced gastric cancer specimens as well as their preneoplastic and non-cancerous counterparts. Alternative CD44 expression was observed in the gastric mucosa with different lesions. CD44 proteins harboring variant exon 6 (CD44 v6) was detected only in cancer tissues with a total positive rate of 14% (10/74). Intracellular RHAMM molecules in Mr 93000 to 95000 were expressed in 3/31 non-cancerous mucosa. RHAMM detection rates increased along with tumor progression. Irrespective of the differences of gross and morphological pattern, majority (54/74) of cancer cases expressed multiple RHAMM isoforms in Mr 40000-45000, 64000, 70000-73000, 85000 and 93000-95000 with the appearance of cell surface immunocytochemical labeling. Among CD44 variant isoforms, v6 is more relevant with malignant transformation of gastric epithelium. Expression of RHAMM, especially the cell surface variants, is closely correlated with tumor progression (P<0.01). Expression of CD44 and RHAMM may benefit the invasion and metastasis of gastric cancer cells presumably in a reciprocal manner. PMID- 11029495 TI - Multiparameter flow cytometry for simultaneous assessment of p53 protein expression and cellular DNA content in oral squamous cell carcinomas: evidence for the development of aneuploid clones from p53-deficient diploid progenitor cells. AB - Diploid tumour cells regularly continue to progress after the development of aneuploid cell populations in head and neck squamous cell carcinomas. The coexistence of aneuploid clones with their diploid progenitor cells provides a unique opportunity to study the order of appearance of p53 mutation and aneuploidy in the same tumour. Multiparameter flow cytometry was therefore applied to 22 oral squamous cell carcinomas to simultaneously assess cellular DNA content and p53 protein expression on a single-cell basis. Concurrent measurements of cytokeratin expression served to identify tumour cells of epithelial origin. One of 5 diploid and 2 of 17 aneuploid carcinomas were p53 negative. For 15 p53-positive aneuploid tumours, overexpression of p53 protein was identified for the aneuploid clones as well as for coexisting diploid tumour cell populations in 14 cases. On the understanding that coexisting diploid and aneuploid tumour cell populations have a common clonal origin, these results provide evidence that aneuploid tumour clones typically develop from p53 deficient diploid progenitor cells. Loss of wild-type p53 function may therefore contribute to the development of aneuploidy in head and neck cancer. PMID- 11029496 TI - The expression of vascular endothelial platelet endothelial cell adhesion molecule-1 is not regulated by IFN-gamma treatment of C6 tumors in vivo. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1) is a glycoprotein that is constitutively expressed on the surface of endothelial cells, leukocytes, platelets, monocytes and neutrophils. It has been proposed that PECAM-1 plays a role in transendothelial leukocyte trafficking. In vitro studies have suggested that interferon-gamma (IFN-gamma) may regulate PECAM-1 expression on endothelial cells. We have investigated the in vivo regulatory role of mouse IFN-gamma (mIFN gamma) on the expression of PECAM-1 on vascular endothelial cells during tumor angiogenesis. In situ retroviral gene transfer was used to deliver mIFN-gamma into established subcutaneous tumors in athymic (nu/nu) mice. The biological activity of mIFN-gamma expressed intratumorally, was verified by i) a macrophage and granulocyte (predominantly neutrophil) infiltrate observed within C6 tumors following mIFN-gamma treatment; ii) an up-regulation of major histocompatibility complex (MHC) class I and II expression on the surface of tumor cells following mIFN-gamma treatment. Indirect immunohistochemical localisation of PECAM-1 expression was performed on control and mIFN-gamma-treated C6 tumor sections. The intensity and distribution of PECAM-1 expression on vascular endothelial cells in mIFN-gamma-treated tumors was similar to control tumors. Immunohistochemical staining of PECAM-1 was compared to factor VIII-related antigen in serial tumor sections. No differences were observed between the vascular endothelial cells expressing PECAM-1 and factor VIII in serial sections of control and mIFN-gamma treated tumors. Therefore, in this in vivo C6 tumor model, IFN-gamma does not alter PECAM-1 expression on tumor-associated vascular endothelial cells. PMID- 11029497 TI - Expression of cell cycle regulator genes in KB, a human squamous cell carcinoma cell line, after irradiation. AB - DNA damage induced by irradiation causes overexpression of the p53 gene, and subsequently the upregulation of p53 downstream genes involved in cell cycle modification. Irradiated malignant cells which possess wild-type p53 have been known to undergo G1 arrest due to p21/Cip1/Waf1 upregulation. Other p53 downstream genes related to the modification of the cell cycle such as gadd45 may cause G2 arrest. Many of the genes which regulate the cell cycle progression have been identified, including the G1 phase specific ink4 family of cyclin-dependent kinase inhibitors (CDK-I), another group of CDK-Is, which affect the cyclin-CDK complexes ubiquitously, and S/G2 accelerator genes. The sequential changes in these cell cycle regulator genes after irradiation has not been clarified. We analyzed the appearance of the apoptotic fraction and cell cycle perturbation after irradiation using KB, a human squamous cell carcinoma line derived from oral floor, and examined the alteration of gene expression for cell cycle regulator genes. The KB cells proceeded to undergo apoptosis in a time and dose dependent manner after irradiation and showed G2 arrest accompanied by upregulation of p53, ubiquitous CDK-Is, and S and G2 accelerator genes. PMID- 11029498 TI - X-ray-induced damage repair in exponentially growing and growth arrested confluent poly(adenosine diphosphate-ribose) polymerase-deficient V79 chinese hamster cell line. AB - We studied the role of PARP in X-ray-induced damage repair using V79 Chinese hamster cells and two derivative cell lines ADPRT54 and ADPRT351 deficient in poly(adenosine diphosphate-ribose) polymerase (PARP) activity. We previously demonstrated that these PARP-deficient cells had drastically reduced levels of p53. Further, these cells were also deficient in downstream endpoints of p53 signaling. In the present study we showed that exponentially growing ADPRT54 and ADPRT351 were hypersensitive to X-radiation compared to the parental V79 cells. Under this condition of growth, although the parental V79 cells exhibit G1 arrest in response to X-irradiation, the PARP-deficient cells do not undergo this specific p53-dependent cell cycle arrest. In contrast, all the cell lines showed similar sensitivity to X-radiation under growth arrested conditions. Further, all the cell lines were equally proficient in performing potentially lethal damage repair (PLDR). Our findings suggest that: i) PARP is involved in X-ray-induced damage repair in replicating cells; ii) PARP is not required for X-ray-induced damage repair in quiescent cells; iii) PARP does not participate in PLDR; iv) deficiency of PARP may potentiate the cytotoxicity of X-irradiation by interfering with the p53-dependent G1 block that occurs after X-irradiation. These results suggest the intriguing possibility that the approach of inhibition of PARP combined with X-radiation may have therapeutic potential for the treatment of fast growing tumors. However, this approach may not be beneficial for slow growing/quiescent tumors. PMID- 11029499 TI - Patterns of expression of Rb and p16 in astrocytic gliomas, and correlation with survival. AB - The retinoblastoma pathway is a key cell cycle regulatory complex that controls the passage of cells through the G1 checkpoint and is a frequent target of genetic alterations in gliomas. In this study, we examined the expression of Rb and p16 in 170 primary astrocytic gliomas by immunohistochemical techniques, and correlated the expression with overall survival to determine their prognostic value as immunomarkers. There were 130 patients with glioblastoma multiforme (GBM) and 40 with anaplastic astrocytoma (AA). Alterations in the levels of Rb or p16 expression were seen in the majority (>90%) of the gliomas studied. The expression of Rb was completely absent or low in 47.5% of the GBM and 67.5% of the AA. The remainder of the tumors was immunopositive for Rb to varying degrees. Immunoreactivity for p16 was absent in 56% of the GBM and 77.5% of the AA. Kaplan Meier survival plots (log-rank test) and Cox proportional hazards regression analysis, adjusted for age and histology, showed that neither Rb nor p16 expression independently predicted survival. The results of our study suggest that although genetic alterations of Rb and p16 are common in gliomas, immunohistochemical analysis of these markers correlates poorly with prognosis. PMID- 11029500 TI - Inhibition of cell growth and telomerase activity of breast cancer cells in vitro by retinoic acids. AB - The effects of retinoic acid (RA) and its analogs, all-trans RA, 9-cis RA and 13 cis RA, were investigated in human breast cancer MCF-7 cells and immortalized breast epithelial cell line MCF-10A. RA inhibited the telomerase activity of MCF 7 cells in a wide range of concentrations. RA at 10 microM also inhibited the growth of MCF-7 cells in a time-dependent manner. However, no significant growth inhibition was found between untreated control and RA-treated MCF-10A cells. Moreover, a marked inhibition of telomerase activity by RA was detected early in MCF-7 cells (after 24 h of RA treatment), which was preceded by a reduction of hTERT mRNA expression (after 12 h of RA treatment). However, MCF-10A cells showed a reduction of telomerase activity and down-regulation of hTERT after 4 days of RA treatment. Simultaneous changes in hTERT mRNA expression and telomerase activity were found for MCF-10A cells. The expressions of hTR and hTEP1 telomerase component genes were not changed after RA treatment. These results indicate that the anti-breast cancer activity of RA could be mediated by its ability to down-regulate the expression of hTERT telomerase gene. PMID- 11029501 TI - Effect of segmental transcatheter arterial chemoembolization on branched chain amino acids and tyrosine ratio in patients with hepatocellular carcinoma. AB - The effect of segmental transcatheter arterial chemoembolization (TAE) on serum amino acid levels and liver function were studied in 23 patients with HCC associated with hepatitis virus C (22 patients) or alcoholism (1 patient), with compensated liver cirrhosis (Child A 18 patients, Child B 5 patients). Serum levels of branched-chain amino acids (BCAA), tyrosine, branched-chain amino acids to tyrosine ratio (BTR), ammonia, total bilirubin and albumin, and prothrombin times were measured before and after TAE (24 h, 7 and 14 days). The BTR was increased significantly 24 h after TAE (p<0.001) and gradually decreased to pre TAE levels. Serum tyrosine levels decreased at 24 h after TAE (p<0.005) and later increased. Serum BCAA levels increased slightly at 7days after TAE and were decreased at 14 days after TAE. This results indicated that the increased BTR was due primarily to the decreased tyrosine level at 24 h after TAE. Serum ammonia levels gradually decreased after TAE and the prothrombin time and serum levels of total bilirubin and albumin were not significantly changed. In this study, segmental TAE had little influence on liver function, and the BTR unexpectedly increased at 24 h after TAE. These results suggest that segmental TAE has minimal side effects and may have a beneficial effect on amino acid metabolism. PMID- 11029502 TI - Telomere length maintenance in aging and carcinogenesis. AB - Normal somatic cells have a finite number of divisions, a limited capacity to proliferate. Human telomeres, the long DNA TTAGGG repeats at the ends of chromosomes, are considered a molecular clock marker. The gradual and progressive telomere shortening at each replicative cycle is associated, through the activation of pRB and p53 pathways and genomic instability, to the replicative senescence, a non-dividing state and widespread cell death. Activation of telomere maintenance [telomerase; or alternative lengthening of telomeres mechanisms (ALT), or other adaptive responses] can revert this program. Although not completely known, several mechanisms and modulating agents may be able to up and down-regulate telomere length and its maintenance. Chemopreventive therapies for the up-regulation of telomerase activity, able to prolong the life of cell cultures in a phenotypically youthful state, could have important applications in research and medicine. On the contrary the therapeutic down-regulation of telomerase activity may be used in cancer therapy. Telomerase expression per se is not oncogenic, but telomere shortening and maintenance seem to be crucial events in tumor formation. Thus a particular focus has been pointed out relatively to the immortalization of normal or potential pre-cancerous cells. With the extension of life span the probability to get in contact with carcinogens increases, genetic instability, oncogene activation and/or onco suppressor gene inactivation (i.e. p53, pRB, ras): the cancer transformation can be then induced in predisposed cells, depending on their genetic context, by the activation of telomere maintenance. Pharmacological intervention may be able to modulate the rate of living, by increasing life span of few specific target cells, or decreasing it in proliferating . Because of the unknown state of the enormous cell number of the human organism, is it safe to extend the human life span by therapeutic agents? PMID- 11029503 TI - Biological activity of all-trans-retinoic acid with and without tamoxifen and alpha-interferon 2a in breast cancer patients. AB - In addition to suppressing breast cancer cell growth, retinoids potentiate growth inhibition in human breast cancer when tested in vitro and in vivo with tamoxifen and/or interferon. The purpose of this study was to ascertain the biologic effects of all-trans-retinoic acid (ATRA) administered alone and with tamoxifen +/- interferon and to identify the relationship between ATRA plasma concentrations and optimal biological dose (the lowest dose that produces a biological response). Three consecutive groups of 15 patients with locally advanced operable breast cancer were treated, in accordance with good clinical practice (GCP) requirements, with ATRA at 3 dose levels alone or with tamoxifen +/- alpha-interferon 2a at flat doses. After 3 weeks, the tumors were surgically removed. Biological parameters measured at the beginning (in biopsy tissue) and end (in surgical tissue) of the study were compared. The optimal biological dose for ATRA was 15 mg/m2/day. Treatments influenced tumor grade but not cell cycle kinetics (G0-G1 phase) or proliferation (Ki67 levels). ATRA induced progesterone receptors independent of dose level and co-administered drugs, but did not induce estrogen receptors when administered alone. Retinoic acid receptor (RAR)-alpha was not affected by treatment and RAR-alpha was moderately influenced whereas RAR beta (concomitantly with transforming growth factor-beta) was induced in 33% of patients by ATRA alone. ATRA pharmacokinetics were dose- and time-dependent. Neither the ATRA + tamoxifen nor the ATRA + tamoxifen + interferon combinations potentiated the ATRA-induced biological changes. Future studies evaluating the role of RAR-beta as a biological marker of retinoid activity are warranted. PMID- 11029504 TI - Urokinase type plasminogen activator and its receptor regulate the invasive potential of gastric cancer cell lines. AB - To assess the role of urokinase-type plasminogen activator (uPA) and uPA receptor (uPAR) on the invasive potential of cancer cells, in vitro experiments were performed using two human gastric cancer cell lines, NUGC-3 and MKN-28. NUGC-3 cells secreted a higher level of uPA than MKN-28 cells, while the uPAR expression of NUGC-3 cells was lower than that of MKN-28 cells. Both cancer cell lines expressed Met protein and did not express hepatocyte growth factor (HGF). In Matrigel invasion assay, MKN-28 cells demonstrated significantly lower invasion index than NUGC-3 cells. The addition of exogenous uPA significantly increased the invasive activity of MKN-28 cells. The uPA expression in NUGC-3 cells was enhanced by adding conditioned media of fibroblast cells or HGF. These results suggest that uPA promotes the invasive capacity of the uPAR-positive cancer cells, and that stromal cells may play an important role in cancer cell invasion by supplying uPA and/or promoting uPA production. PMID- 11029505 TI - High-dose chemotherapy with tandem autologous transplantation as part of the initial therapy for aggressive non-Hodgkin's lymphoma. AB - The purpose of the present study was to evaluate the feasibility and the efficacy of employing a high-dose chemotherapy (HDT) regimen with tandem peripheral blood progenitor cells (PBPC) supported transplantation in the initial treatment of aggressive non-Hodgkin's lymphoma (NHL). HDT was preceded by a standard course of conventional dose chemotherapy in 17 out of the 25 patients treated, while in 8 cases it was delivered after only one or two cycles. HDT was a three-step procedure which included high-dose (6-7 g/m2) cyclophosphamide (CY) supported by haematopoietic growth factors, the first myeloablative course with mitoxantrone (NOV) 60, 75 or 90 mg/m2 plus melphalan (L-PAM) 140-180 mg/m2 with haematopoietic rescue, and the second myeloablative course with etoposide (VP) and carboplatin (CARBO) given at 1.5 g/m2 each with haematopoietic rescue. PBPC were collected after CY administration. Twenty-two patients (88%) completed the HDT, haematological reconstitution was rapid and complete at each step and there were no toxic deaths. The activity of the treatment was high with a CR rate over 90% in the entire patient population. The 2-year overall survival (OS) and failure free survival (FFS) rates of patients in both Age-Adjusted International Prognostic Index (A-AIPI) groups 2 and 3 are 79% and the disease-free survival (DFS) rate for the CRs is 85%. In A-AIPI group 1 the 2-year OS and FFS rates are both 91%. PMID- 11029506 TI - Galectin-1 gene expression and methylation state in human T leukemia cell lines. AB - Galectin-1 has been demonstrated to be a mediator of T-cell apoptosis acting on activated T-cells and, in a selective manner, on different T leukemia cell lines. Here we show that the sensitivity to galectin-1 is associated with repression of the endogenous galectin-1 gene whereas non-sensitive cells express high levels of galectin-1. Repression of galectin-1 gene in sensitive cells is associated with hyper-methylation of the promoter region. Transient treatment of non-expressing cells with the demethylating agent 5-azacytidine led to irreversible demethylation and subsequent reactivation of galectin-1 gene. PMID- 11029507 TI - The affinity of MCF7 breast cancer cells to hyaluronan substrates of different molecular weight and concentrations in an in vitro model. AB - The affinity of MCF7 breast cancer cells to hyaluronan (HA) was investigated in an in vitro model. The cells form a tightly adhering monolayer on native HA with a concentration of 5 mg/ml. On native HA at higher concentrations the cells reduce their adhesion to the substrate in favor of increased intercellular bonds, resulting in a cluster-like aggregate that tends to detach from the substrate. Aggregate formation is accomplished after 12 h incubation. The phenomenon is independent of the CD44 receptor. Degradation of native HA by hyaluronidase abolishes aggregate formation even at high HA concentrations in favor of formation of a firmly adhering monolayer. This model may help to understand tumor spread on HA tissue structures and may explain therapy successes with hyaluronidase in tumor patients. PMID- 11029508 TI - Patterns of chromosomal imbalances in muscle invasive bladder cancer. AB - Cytogenetic investigations of bladder cancer suggested that development and progression is characterized by specific chromosomal aberrations. In order to identify genetic changes linked to muscle invasive tumors and metastatic growth we analyzed 67 bladder carcinomas (30 pT1 and 37 pT2-4) by means of comparative genomic hybridization (CGH). The most frequent changes were gains of chromosome 1q (54%), 8q (54%), 17q (49%), 2p (30%), 12 (30%), 5p (25%), 3q (24%) and 6p (24%) as well as losses of 11p (43%), 8p (42%), 9p (36%), 11q (34%), 2q, 4q, 5q (30% each), 9q (27%) and 10q (27%). Previously not described amplifications were found at 5p11-p13, 7q21-q31, 9p24 and 17q24-q25. Gains of 3q, 7p, and 18p were markedly more frequent in pT2-4 in comparison to pT1 carcinomas but the difference did not reach statistical significance. Non-metastatic tumors showed more aberrations on average than metastatic carcinomas, although no particular change was found to be predominating in either group. Our data confirm previous findings of strong genetic similarities between minimally and deeply invasive bladder carcinomas but argue for differences between metastatic and non metastatic disease. PMID- 11029509 TI - Mutation profile of the p53, fhit, p16INK4a/p19ARF and H-ras genes in Indian breast carcinomas. AB - Breast cancer is the second most prevalent cancer affecting Indian women. Genetic alterations of oncogenes and tumor suppressor genes were attributed to the development of breast carcinomas. In the present study, human breast tumor DNAs from untreated, non-familial, Indian patients were analysed for the presence of mutations in p53, fhit, p16INK4a/p19ARF and H-ras genes. Polymerase chain reaction-single strand conformation polymorphism and sequencing analysis were used to detect point mutations. Exons 5-8 of p53, exons 1-2 of p16INK4a, exon 2 of p19ARF, exons 5-9 of fhit gene and exons 1-2 of H-ras genes were amplified and analysed individually using exon-flanking primers. Only 12% of the tumors had mutation in p53, 8% had mutation in fhit gene and none of the tumors showed evidence for mutation in p16INK4a/p19ARF and H-ras genes. Tumor B18 exhibited two novel mutations in the p53 gene, ATGright curved arrow GTG (Metright curved arrow Val) at codon 237 and AATright curved arrow GAT (Asnright curved arrow Asp) at codon 263. Both of these mutations are hitherto unreported in breast carcinomas. Tumor B20 had a non-sense mutation CGAright curved arrow TGA (Argright curved arrow Stop) at codon 306 of p53 gene. In fhit gene, tumor B1 exhibited TTCTright curved arrow TACT mutation at intron 8 and tumor B15 had a silent mutation GAGright curved arrow GAA (Gluright curved arrow Glu) at codon 123. Our results indicate that, among the genes analysed, the p53 gene was more frequently mutated than fhit, p16INK4a/p19ARF and H-ras genes in Indian mammary tumors. Transcribable point mutations of fhit gene were found to be extremely uncommon in these tumors. Mutations in the above genes are mutually exclusive and are infrequent in fhit, p16INK4a/p19ARF and H-ras genes suggesting that these genes may not play a major role in Indian breast carcinomas. However, the significant frequency of mutations in the p53 gene suggest that p53 could be one of the genes involved in the genesis of sporadic breast carcinomas in Indian women. PMID- 11029510 TI - New aromatase inhibitors in the treatment of advanced breast cancer. AB - New aromatase inhibitors are an exciting treatment option for postmenopausal women with hormone sensitive breast cancer. They have been shown to reduce tumors in a significant number of patients, and exhibit definite antitumor activity at a relatively low daily dose, and are highly potent, highly selective, and well tolerated. Results from recent clinical phase III studies have confirmed their efficacy and the key role they have in the therapy for advanced breast cancer in postmenopausal women. The agents available for clinical use are: letrozole, anastrozole, and exemestane. These drugs have demonstrated high activity in women failing tamoxifen in locally advanced or metastatic disease. This communication reviews the clinical use of aromatase inhibitors, particularly in second and first line hormonal treatment of advanced breast cancer. PMID- 11029511 TI - UCN-01 and camptothecin induce DNA double-strand breaks in p53 mutant tumor cells, but not in normal or p53 negative epithelial cells. AB - Previous research has shown synergistic growth inhibition between UCN-01 and camptothecin (CPT) in tumor cells with mutant p53 versus tumor cells with wild type p53. To determine the possible role of p53 in this drug combination, we tested the hypothesis that the synergistic growth inhibition is due to the absence of p53, and can result from the induction of DNA double-strand breaks (DSBs). Experiments were performed with the use of normal human mammary epithelial cells (HMEC); HMEC transfected with HPV16 E6 protein which inactivates p53 (HE6), or p53-mutant MDA-MB-231 tumor cells. CPT, UCN-01, or a 1:1 combination of both, in either HMEC or HE6 cells did not induce DSBs. In contrast, simultaneous treatment of MDA-MB-231 cells with both UCN-01 and CPT induced significant levels of DSBs while treatment with either drug alone did not. While UCN-01 was surprisingly potent against HMEC, the growth inhibition was only additive between UCN-01 and CPT against these cells. HE6 cells were much less sensitive than HMEC to UCN-01 and slightly less sensitive to the combined treatment with UCN-01 and CPT. The drug combination was synergistic against HE6 cells, due to their lower sensitivity to UCN-01. Unlike what was observed previously in MDA-MB-231 cells, UCN-01 did not abrogate CPT-induced inhibition of DNA synthesis in either HMEC or HE6 cells. These data indicate that synergistic growth inhibition by UCN-01 and CPT against p53 mutant MDA-MB-231 tumor cells may be due to induction of DSBs however the loss of p53 function alone does not sensitize normal cells to the combination of both drugs. PMID- 11029512 TI - Opioid growth factor regulates the cell cycle of human neoplasias. AB - The native opioid growth factor (OGF), [Met5]-enkephalin, is a tonic inhibitory peptide that modulates cell proliferation and migration, as well as tissue organization, during development, cancer, homeostatic cellular renewal, wound healing, and angiogenesis. OGF action is mediated by the OGF receptor (OGFr). To investigate the target of OGF as to cell proliferation, the effects of excess OGF, and a deprivation of OGF-OGFr interaction by an opioid antagonist, naltrexone (NTX), were examined in 3 human cancer cell lines: pancreatic (BxPC 3), colon (HT-29), and head and neck (CAL-27). OGF exposure decreased growth, DNA synthesis, and mitosis, and increased the doubling time from control levels. FACS analysis revealed a marked increase in cells in the G0/G1 phase and compensatory reduction in cells in S and G2/M phases. Consistent with this observation, the percentage of labeled mitosis (PLM) analysis showed a notable increase in the time of the G0/G1 phase. Receptor blockade with NTX increased the rate of growth, length of DNA synthesis and mitotic phases, and decreased doubling time from control values. FACS analysis indicated an increase in the proportion of cells in S and G2/M phases, and a decrease in the number of cells in the G0/G1 phase. PLM evaluation demonstrated a shortening of the length of the S and G2 phases in the 3 cell lines, and decreases in the M and G0/G1 phases in some cancers. These results indicate that OGF action is directed at the G0/G1 phase, but interruption of OGF-OGFr interfacing has widespread repercussions on the cell cycle. The data on blockade of OGF-OGFr during log phase growth suggest a requisite escorting of the growth peptide and its receptor through the cell cycle. PMID- 11029513 TI - Receptor mediated antiproliferative effects of the cytotoxic LHRH agonist AN-152 in human ovarian and endometrial cancer cell lines. AB - Eighty percent of human ovarian and endometrial cancers express receptors for luteinizing hormone-releasing hormone (LHRH). These receptors might be used for targeted chemotherapy with cytotoxic LHRH analogs such as AN-152, in which doxorubicin is linked to agonist carrier [D-Lys6]LHRH. The antiproliferative effects of doxorubicin and AN-152 were assessed in LHRH receptor-positive ovarian (EFO-21, EFO-27) and endometrial (HEC-1A, Ishikawa) cancer cell lines as well as in LHRH receptor negative ovarian SKOV-3 and endometrial MFE-296 lines. The mechanism of action of AN-152 was investigated by a blockage of receptors using an excess of the LHRH agonist [D-Trp6]LHRH. In some cases, confocal laser scanning microscopy was used to visualize the accumulation of AN-152 or doxorubicin within the cells. In 3 of 4 LHRH receptor-positive cell lines (EFO 21, HEC-1A, Ishikawa) AN-152 was more effective than doxorubicin in inhibiting cell proliferation. The effect of AN-152 was shown to be receptor-mediated because it could be reduced by competitive blockade of the LHRH receptors with [D Trp6]LHRH. In contrast, AN-152 was less active than doxorubicin in LHRH receptor negative lines. Confocal laser-scanning microscopy showed an intranuclear accumulation of AN-152 and competitive inhibition thereof by [D-Trp6]LHRH in LHRH receptor-positive cell lines, but no intracellular accumulation of AN-152 could be detected in the receptor-negative SKOV-3 line. These results suggest a selective receptor-mediated action of AN-152 in receptor-positive cell lines. PMID- 11029514 TI - Regulation of tumor cell apoptotic sensitivity during the cell cycle (Review). AB - Understanding how current chemotherapeutic modalities induce apoptosis is critical to designing better anti-cancer agents. This review is concerned with how pharmacological agents induce tumor cell apoptosis in a cell cycle-dependent manner. Recent experiments demonstrate that expression of several apoptotic regulatory proteins (such as Bcl-2, Bax, p53, and Survivin) are differentially regulated according to the phases of the cell cycle. This cell cycle-dependent regulation in turn contributes to increased drug sensitivity in different phases of the cell cycle. It is therefore likely that the cell cycle-dependent expression of cell death proteins plays a role in regulating chemosensitivity and apoptotic commitment of human tumor cells. PMID- 11029515 TI - Recombinant immunotoxins for the treatment of Hodgkin's disease (Review). AB - In recent years, substantial experience has been accumulated with tumor-specific immunotherapeutics which seem to be effective against minimal residual disease. The coupling of toxins to monoclonal antibodies has indicated promising results in early clinical trials. Recombinant DNA technology makes it possible to genetically fuse coding regions of V genes or cytokines to modified toxin domains. These recombinant immunotoxins can easily be manipulated to increase the cytotoxic potency or affinity. Binding single-chain variable fragments (scFv) expressed as chimeric fusion proteins on the surface of filamentous bacteriophages were selected on Hodgkin-derived cell lines. This technique was also used to create a new humanized anti-CD30 scFv which exhibits similar binding to the CD30 antigen when compared to its murine predecessor. ScFvs were then inserted into a new bacterial expression vector and thus fused to a deletion mutant of Pseudomonas exotoxin. Anti-CD25(scFv)-ETA' and anti-CD30(scFv)-ETA' were isolated from E. coli periplasm and purified by metal chelate affinity and size exclusion chromatography. All immunotoxins produced showed specific cytotoxicity against Hodgkin lymphoma cell lines as documented by competitive assays. In addition, these constructs were highly efficient in the treatment of disseminated human Hodgkin's disease in SCID mice. These in vivo data indicate a possible clinical impact for patients with relapsed CD25- and/or CD30-positive lymphoma. PMID- 11029516 TI - Autologous platelet-derived wound healing factor promotes angiogenesis via alphavbeta3-integrin expression in chronic wounds. AB - Healing of venous leg ulcers depends on the adhesive interaction and formation of new vascular cells. Angiogenesis on the surface of angiogenic blood vessels requires the vascular integrin alphavbeta3 also known as the vitronectin receptor. Autologous platelet-derived wound healing factor (autologous PDWHF) has been described to regulate the wound healing process by forming granulation tissue in the early healing phase. Here we analysed the influence of autologous PDWHF on the expression of the alphavbeta3 integrin in tissue specimen of venous leg ulcers in comparison with placebo treated controls by using reverse transcriptase-polymerase chain reaction and immunohistochemistry. Our investigations provide evidence that mRNA and protein expression of alphavbeta3 were significantly increased in healing venous leg ulcers after 96 h treatment (p<0.05), whereas the total amount of alphavbeta3 mRNA and protein was not altered in placebo treated patients. In healing leg ulcers the alphavbeta3 integrin was predominantly localized around capillary vessels preferentially at sites of newly formed granulation tissue. Placebo controlled patients displayed no altered expression of the alphavbeta3 integrin in biopsy specimen. These findings suggest that topical autologous platelet-derived wound healing factor influences the process of angiogenesis/revascularization via alphavbeta3 integrin expression hereby promoting granulation tissue formation in healing leg ulcers. PMID- 11029517 TI - Curcumin inhibits lipoxygenase by binding to its central cavity: theoretical and X-ray evidence. AB - Many lipoxygenase inhibitors including curcumin are currently being studied for their anti-carcinogenic properties. Curcumin is a naturally occurring polyphenolic phytochemical isolated from the powdered rhizome of the plant Curcuma longa that possesses anti-inflammatory properties and inhibits cancer formation in mice. Recently it was shown that the soybean lipoxygenase L1 catalyzed the oxygenation of curcumin and that curcumin can act as a lipoxygenase substrate. In the current study, we investigated the fate of curcumin when used as a soybean lipoxygenase L3 substrate. By use of X-ray diffraction and mass spectrometry, we found an unoccupied electron mass that appears to be an unusual degradation product of curcumin (4-hydroxyperoxy-2-methoxyphenol) located near the soybean L3 catalytic site. Understanding how curcumin inhibits lipoxygenase may help in the development of novel anti-cancer drugs used for treatment where lipoxygenases are involved. PMID- 11029518 TI - Cyclooxygenase expression in the gallbladder. AB - The COX expressions were evaluated separately in the epithelium and in the stroma of gallbladder cancer, chronic cholecystitis, xanthogranulomatous cholecystitis (XGC) and the normal gallbladder. In normal gallbladder COX-2 expression rate was significantly higher in the epithelium than in the stroma. The COX-2 expression rate in the epithelium of non-cancerous adjacent epithelium to cancerous lesion was significantly lower than those not only of cancer, but also chronic cholecystitis, XGC and normal gallbladder. In stroma, the COX-2 expression rate in cancer, chronic cholecystitis and XGC were significantly higher than that of the normal gallbladder. The rate in non-cancerous adjacent stroma to cancer is significantly lower than that of cancer and XGC. However, the difference of rate between of normal and of chronic cholecystitis was not significant. The COX-2 expression rates were significantly higher in both the epithelium and the stroma in the well and moderately differentiated cancer group than in the poorly and undifferentiated cancer group. Our results suggest that COX-2 expression in the gallbladder may be regulated by various factors and not directly related to carcinogenesis. The significance of its repression in the non-cancerous adjacent tissue to cancer lesion should be re-evaluated. PMID- 11029519 TI - Impaired Ca2+-ATPase oligomerization and increased phospholamban expression in dilated cardiomyopathy. AB - Although primary genetic defects have been identified for some forms of inherited cardiomyopathy, it is not well understood how secondary abnormalities actually lead to muscle cell destruction. Since cardiomyopathies significantly influence morbidity and mortality rates world-wide, it is important to improve the differential diagnosis of these disorders and develop potential treatments for inherited diseases of the heart. Elucidation of the secondary molecular mechanisms underlying cardiac cell necrosis might help linking a specific mutation in a cardiac gene to acute heart failure. As disturbed Ca2+-homeostasis may contribute to heart failure, we have investigated the relative abundance and oligomeric status of the sarcoplasmic reticulum Ca2+-ATPase and phospholamban in various cardiomyopathies. These two proteins represent important factors in cardiac relaxation. The SERCA2 isoform of the Ca2+-ATPase represents a major Ca2+ removal system in cardiac muscle fibres and phospholamban is a regulator of Ca2+ pump activity. Although Ca2+-ATPase expression did not seem to be markedly altered, the comparative immunoblot analysis presented here clearly shows that phospholamban expression is increased in dilated cardiomyopathy, possibly explaining the decreased Ca2+-uptake in the disease. In contrast to the normal enzyme, the Ca2+-pump was demonstrated to exhibit an impairment of crosslinker stabilized oligomerization in dilated cardiomyopathy. Since Ca2+-ATPase oligomerization is important for co-operative kinetics and protection against proteolytic degradation, the monomeric Ca2+-ATPase may trigger an abnormal contraction-relaxation cycle in dilated cardiomyopathy leading to heart failure. PMID- 11029520 TI - The role of S100A4 gene encoding an S100-related calcium-binding protein in human bile duct adenocarcinoma cell lines: correlation of S100A4 expression and invasive growth in Matrigel Matrix. AB - S100A4, one of the tandemly arranged S100 genes at chromosome 1q21, has been suggested to play a functional role in cell motility and invasiveness of tumor cell growth. We investigated the expression of S100A4 and the in vitro invasiveness of 4 human bile duct adenocarcinoma cell lines by the Matrigel assay. S100A4 was abundantly expressed in 2 adenocarcinoma cell lines whose growth pattern was highly invasive. Induction of antisense S100A4 into one of the cell lines decreased S100A4 mRNA levels and reduced invasiveness. In contrast, induction of sense S100A4 expression into non-invasive KMBC adenocarcinoma cells, which originally lacked S100A4 expression, resulted in apparent invasive potential in the transfected cells compared with the cells with the vector alone. These results suggest that S100A4 expression is well correlated with the invasiveness of human bile duct adenocarcinomas. PMID- 11029521 TI - Trypsinogen expression in colorectal cancers. AB - The purpose of this study was to evaluate the trypsinogen expression in human colorectal cancers. We observed the trypsinogen-1 mRNA expression in five of ten human colorectal cancer cell lines by reverse transcriptase-polymerase chain reaction (RT-PCR), and gelatinolytic activity in conditioned medium of cancer cells at acidic pH by gelatin zymography. Furthermore, trypsinogen protein expression was observed in 68 out of 154 (44.2%) surgical specimens of colorectal cancer immunohistochemically. However, there was no statistically significant relationship between the trypsinogen expression and the clinicopathological findings. These findings suggest that trypsinogen might be involved in cancer invasion and metastasis throughout the cancer progression. PMID- 11029522 TI - Fate of 2-deoxy-2-[18F]fluoro-D-glucose in hyperglycemic rats. AB - Sustained hyperglycemia allows the preferential labelling of pancreatic glycogen by D-[U-14C]glucose in control rats, as compared to animals previously injected with streptozotocin (STZ rats). The major aim of the present study was to investigate whether a sizeable difference between control and STZ rats could also be observed in terms of the radioactive content of the pancreatic gland 8 h after the intravenous injection of 2-deoxy-2-[18F]fluoro-D-glucose, both types of animals being examined at the same level of hyperglycemia. Although the radioactive content of muscle, liver and kidney was lower in STZ rats than in control animals, such a difference failed to achieve statistical significance in brain, hypophysis, pancreas and parotid gland. It is proposed, therefore, that 11C-labelled D-glucose, rather than 2-deoxy-2-[18F]fluoro-D-glucose should be used in the perspective of the non-invasive imaging of the endocrine pancreas. PMID- 11029523 TI - Caspase-1 activity as a possible predictor of apoptosis induced by cisplatin in gastric cancer cells. AB - Recent studies have shown that caspases, which are cystein proteases, elevate endonuclease activity and induce apoptosis. Caspase-1, an interleukin-1beta converting enzyme, has been reported to be related with anti-cancer drug induced apoptosis as well as with caspase-3. To elucidate the caspase-1 activity, which might be a predictor for the effect of chemotherapy, we examined the changes of caspase-1 activity induced after exposure to cisplatin (CDDP) in six gastric cancer cell lines. A high correlation between the 50% inhibitory concentration (IC50) and caspase-1 activity ratio was shown (r=0.83, p=0.041) (caspase-1 activity ratio: the caspase-1 activity of cells at 4 h after CDDP treatment/the caspase-1 activity of untreated cells). Further, we examined the correlation between caspase-1 activity and apoptosis induced by CDDP in two cell lines that have very different CDDP sensitivities; OCUM-2M and OCUM-2M/DDP (IC50; 0. 85+/ 0.4 microg/ml and 9.0+/-1.2 microg/ml, respectively). The apoptotic index of OCUM 2M was significantly higher than that of OCUM-2M/DDP (19.8+/-3.8% vs. 4.5+/-1.2%, respectively; p=0.0005). In both cell lines, caspase-1 activity began to increase immediately after exposure to CDDP and peaked at approximately 4 h after cessation of exposure to CDDP, and gradually decreased thereafter. The caspase-1 activity of OCUM-2M was approximately 1.8-times higher than that of OCUM-2M/DDP at 4 h after exposure to CDDP. Taken together, our results indicate that evaluating the changes of caspase-1 activity after exposure to CDDP may be useful to predict apoptosis following CDDP treatment in gastric cancer cells. PMID- 11029524 TI - Growth inhibitory effect on glioma cells of adenovirus-mediated p16/INK4a gene transfer in vitro and in vivo. AB - The tumour suppressor gene p16/INK4a encodes a specific inhibitor of the cyclin D dependent kinases CDK4 and CDK6. p16/INK4a prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumour suppressor protein (pRb), thus preventing exit from the G1 phase. In human cancers, the estimated frequency of genetic alteration involving the p16/INK4a locus is believed to be second only to alteration of p53. A high frequency (greater than 50%) of homozygous p16/INK4a gene deletion has been demonstrated in glioblastoma tissues and p16/INK4a is altered in 80% of glioma cell lines. Therefore, restoration of p16/INK4a would suppress cell proliferation and induce cell growth arrest. We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16/INK4a induced growth suppression in vitro and in vivo. Expression of p16 transferred by Ad-CMV-p16/INK4a in glioma cells was highly efficient and maintained for more than seven days. In addition, we found that the endogenous status of p16 and Rb might affect the expression of exogenous p16/INK4a gene and inhibitory effect of cell proliferation. Even though, there were several factors affecting the efficiency of Ad-CMV-p16/INK4 gene transfer, our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas. PMID- 11029525 TI - Patterns of allelic loss at the BRCA1 locus in Arabic women with breast cancer. AB - Loss of heterozygosity (LOH) of BRCA1, a tumor suppressor gene, is one mechanism of genetic inactivation in both sporadic and familial forms of breast cancer. Studies reported in breast cancers from women of Northern European descent have shown LOH in 30-50% of sporadic tumors. Microsatellite instability (MSI) has served as evidence for involvement of DNA repair genes. This study investigates the extent of allelic imbalance at the BRCA1 region in Arabic women with breast cancer. Paired normal and tumor tissue were available for DNA analysis in 13 cases. Results using fluorescent tagged primers to microsatellite markers D17S1323, D17S1325 and D17S855 intragenic to BRCA1 were analyzed using an ABI 310 DNA sequencer. As compared to normal DNA, MSI and LOH were recognized as a gain and a loss, respectively, of one signal in one allele in the tumor DNA. Microsatellite analyses showed 12 of 13 (92%) cases with LOH or MSI or both. Three cases demonstrated LOH alone, 3 cases with MSI alone. Six cases indicated both LOH and MSI; 2 cases with either LOH or MSI in separate markers. The combined finding of LOH and MSI in the same marker was detected only with D17S1325 in 4/6 cases. The proportion of aberrant findings of the BRCA1 locus in breast cancer appears to be higher in Arabic women than in other populations studied to date. PMID- 11029526 TI - HCV and hepatocellular carcinoma: A case-control study in an area of hyperendemicity. AB - HCV is an RNA virus that cannot be integrated with the host genome; it can, however, exert its oncogenetic potential indirectly by contributing to the modulator effects of the host immune system probably through a capacity to escape the immune system. We have carried out a case controlled study in a hyperendemic area on HCV infection and liver cancer. We screened 114 liver cancer and 226 controls. All patients were at first diagnosis and examined. For liver cancer the risk was (OR=32.9, 95% CI 16.5-65.4, p<0.0001). Our study is particularly important for public health since it shows that in the South of Italy, because of the high prevalence of HCV and the high life expectancy, there are good reasons to suppose that the incidence rate of liver cancer will continue to increase in the next few years. PMID- 11029527 TI - Two BCG vaccine formulations prepared from the same strain with different J774 macrophage activation capacities and patterns of NF-kappaB induction. AB - Two BCG vaccine formulations of the Moreau strain, commercially manufactured for anti-tuberculosis vaccination, ID-BCG, or anti-cancer adjuvant therapy, Onco-BCG, were compared for immunogenic activity in vitro. The growth rates of both vaccines in murine macrophages were the same, however, Onco-BCG induced stronger and longer-lasting secretion of TNF-alpha, IL-6 and nitric oxide. Onco-vaccine was also more potent in inducing NF-kappaB p65/p50 DNA-binding activity whilst in ID-BCG-infected cells the activity was transient and then gradually replaced by the transcriptionally inactive homodimer p50/p50. Comparative analysis of mycobacterial antigens of the two vaccines demonstrated a difference in expression of the 19 kDa and 38 kDa lipoproteins detected only in Onco-BCG extracts. These results suggest that these molecules may be responsible for the vigorous activation of macrophages induced by the Onco-vaccine. The data obtained show that vaccines from the same BCG strain, when manufactured differently, can vary significantly in their antigen expression and, consequently, in their capacity for macrophage activation which could contribute to the difference in their immunopotentiating effects. PMID- 11029528 TI - Expression of Fas and Fas ligand reflects the biological characteristics but not the status of apoptosis of intrahepatic cholangiocellular carcinoma. AB - Although intrahepatic cholangiocellular carcinoma (CCC) is the second most common of hepatic malignancies, there have been few studies evaluating its biological characteristics. We therefore decided to investigate the status of apoptosis and of Fas and Fas ligand expression in this carcinoma. We performed immunohistochemistry for Fas and Fas ligand in 40 CCC cases and evaluated the apoptotic index (AI) by counting the number of morphologically apoptotic cells per 1000 cells. AI was higher in cases with poor differentiation, lymph node metastasis and high Ki-67 labeling index (LI). Fas expression in CCC cells decreased in cases with poor differentiation and high Ki-67 LI. Fas ligand expression in the stromal infiltrating mononuclear cells did not correlate with any clinicopathological features of CCC, but was directly related to Fas expression in CCC cells. Fas ligand was also expressed in CCC cells in 27.5% of the cases and more frequently observed in cases with moderate or poor differentiation and high Ki-67 LI. None of these three parameters correlated with AI. These findings indicate that, in CCC, i) cases showing rapid growth characteristics are less likely to die of Fas-mediated apoptosis and more likely to display counterattack to lymphocytes via Fas ligand expressed by carcinoma cells; ii) stromal mononuclear cells express Fas ligand in response to Fas expression in carcinoma cells; iii) Fas and Fas ligand expression are not related to the status of apoptosis. PMID- 11029529 TI - No association between an intronic biallelic polymorphism of the FE65 gene and Alzheimer's disease. AB - The cleavage of the amyloid precursor protein (APP) into amyloidogenic components (Abeta) is a central event in the pathogenesis of Alzheimer's disease (AD). FE65 is a protein that is involved in APP metabolism and may facilitate the production of Abeta. Recently, an intronic polymorphism of the gene encoding FE65 (FE65) was associated with altered risk for the development of sporadic AD. In our sample of 102 AD patients and 351 non-demented controls we did not replicate the association between FE65 and AD. Moreover, we observed no risk-modifying interaction and no linkage disequilibrium between FE65 and the gene encoding the acid protease cathepsin D (catD), which - like FE65 - is involved in APP metabolism and is also located on chromosome 11p15. We conclude that, whereas FE65 is implicated in AD pathology, the gene encoding FE65 does not appear to confer a substantial risk for AD. PMID- 11029530 TI - The influence of angiotensin converting enzyme inhibitors on lipid peroxidation in sera and aorta of rabbits in diet-induced hypercholesterolemia. AB - In hypercholesterolemia increased lipid and lipoprotein peroxidation occurs. The renin-angiotensin system plays an important role in atherogenesis. Angiotensin II induces smooth muscle cells proliferation and stimulates oxidation of LDL particles and foam cell accumulation. Inhibition of ang II production leads to decrease in lipid peroxide production. The aim of this study was to assess the lipid peroxidation expressed as concentration of thiobarbituric acid reactive species (TBARS) in sera and aorta homogenates after administration of two doses of angiotensin-converting enzyme (ACE) inhibitors (captopril, enalapril and quinapril) in diet-induced hypercholesterolemia in rabbits. Sixty-four New Zealand rabbits were used. Animals were fed with standard fodder, special diet (1% cholesterol content) or special diet + tested ACEI. Two doses of ACE inhibitors were used: i), equivalent to applied to humans, ii), dose 10 times higher. The animals were divided into 8 groups: control, standard fodder; B, special diet; C1, C2, special diet + captopril in doses 2.5 and 25 mg/kg/24 h, respectively; E1, E2, special diet + enalapril in doses 0.75 and 7.5 mg/kg/24 h, respectively; Q1 and Q2, special diet + quinapril in doses 0.75 and 7.5 mg/kg per day, respectively. In cholesterol-fed rabbits and in groups receiving lower doses of tested ACE inhibitors, the serum TBARS concentration at 6 months was significantly higher in comparison to the control. The higher doses of enalapril, quinapril and captopril, prevented the cholesterol-induced rise in TBARS concentration. Lower dose of captopril attenuated the rise in TBARS concentration, it was significantly lower in comparison to group B, but higher than in the control group. In animals from groups B, E1, C1, Q1 TBARS concentration in aortae was significantly higher as compared to control group. Both doses of captopril and higher doses of enalapril and quinapril inhibited the rise of lipid peroxides concentration induced by cholesterol-rich diet. PMID- 11029531 TI - Soluble CD44 splice variants and pelvic lymph node metastasis in ovarian cancer patients. AB - Alternative splicing of CD44 and aberrant levels of soluble CD44 protein in the serum of cancer patients has been correlated to tumor progression and metastasis. To examine the clinical value of CD44 serum levels (sCD44) in ovarian cancer we determined concentrations of the soluble, variable isoforms sCD44std, sCD44v5 and sCD44v6 with a sensitive ELISA. Pre-operative serum samples from 66 patients with histologically diagnosed invasive disease as well as sera taken from 40 healthy blood donors were analyzed. In sera of ovarian cancer patients we detected elevated concentrations of overall CD44 serum levels represented by sCD44std (p=0.001), but decreased levels of the specific isoforms CD44v5 (p=0.0002) and v6 (p=0.0001). This is the first report demonstrating that ovarian cancer patients with pelvic lymph node metastasis at the time of diagnosis showed specifically elevated sCD44v6 (p=0.073) serum concentrations in comparison to patients without lymph node involvement, whereas overall sCD44 serum levels did not differ. Decreased serum levels of sCD44v5 were found in progesterone receptor-positive tumors (p=0. 059) and postmenopausal patients (p=0.032). Increased concentrations of sCD44v6 were detectable in estrogen receptor-positive tumors but not significantly (p=0.138). Serum CD44v5 levels were associated with shortened relapse-free survival time. No association was found between serum CD44 isoforms and the classical clinicopathological parameters stage and grading or overall survival. CD44 splice variants are possibly involved in a complex interaction with the hormonal environment during tumorigenesis and metastasis of ovarian cancer. PMID- 11029532 TI - Calbindin-D28k immunoreactivity in the suprachiasmatic nucleus and the circadian response to constant light in the rat. AB - Recent studies in the hamster have led to the discovery that the expression of the calcium binding protein, calbindin-D28k, is a defining feature of neurons in the suprachiasmatic nucleus involved in the regulation of circadian rhythms by environmental light.(2,18, 19,32) To study further the involvement of calbindin D28k, we examined the effect of exposure to constant light on calbindin-D28k immunoreactivity in the suprachiasmatic nucleus of intact rats and of rats treated neonatally with the retinal neurotoxin, monosodium glutamate. Exposure to constant light is known to disrupt circadian rhythms in rodents and we found previously that treatment with monosodium glutamate selectively prevents the disruptive effect of constant light on circadian rhythms in rats.(7,9) In the present study we found that exposure to light suppresses calbindin-D28k expression in the ventrolateral retinorecipient region of the suprachiasmatic nucleus of rats and that neonatal treatment with monosodium glutamate blocks the suppressive effect of constant light on calbindin-D28k expression. These findings are consistent with the proposed role of calbindin-D28k in photic signaling in the suprachiasmatic nucleus,(32) and point to the possibility that suppression of calbindin-D28k expression is linked to the mechanism by which constant light disrupts circadian rhythms. PMID- 11029533 TI - Age-related alteration of the forepaw representation in the rat primary somatosensory cortex. AB - The cortical forepaw representation of adult rats was mapped by using multiunit recordings of layer IV neurons within the primary somatosensory cortex. Electrophysiological maps were based on somatosensory "submodality" (cutaneous vs non-cutaneous), location and size of the receptive fields. Age-related changes in the organizational features of the forepaw representation in the primary somatosensory cortex were analysed in adult rats whose ages ranged from 3.5 to five months (young), about eight months (mature), 15-17months (presenescent) to 24-28months (senescent). Rats were housed from weaning (30days postnatal) in a standard laboratory environment. The organization of the forepaw map was not gradually altered with advancing age, but striking changes occurred in early adulthood (before eight months) and did not progress with further aging. The main alterations consisted of a prominent decrease in, and a fragmentation of, the cutaneous area of the forepaw representation. Representational zones formerly serving cutaneous surfaces became predominantly activated by high-threshold, presumably non-cutaneous, inputs which appeared somatotopically organized. These emergent non-cutaneous zones were interspersed with cutaneous sectors, thereby disrupting the somatotopic organization of the map of the forepaw skin. No significant modification in the size of glabrous or hairy cutaneous receptive fields accompanied these changes. Subjective evaluation of the responses evoked by tactile stimulation suggests that neuronal responsiveness was increased in the eight- to 17-month-old rats, but less so in the 24- to 28-month-old animals. These results indicate that degradation of the somatotopic organization of the cutaneous representation of the forepaw in the rat somatosensory cortex occurs early during the course of adult life. PMID- 11029534 TI - Laminar differences in recurrent excitatory transmission in the rat entorhinal cortex in vitro. AB - Paired intracellular recordings were used to investigate recurrent excitatory transmission in layers II, III and V of the rat entorhinal cortex in vitro. There was a relatively high probability of finding a recurrent connection between pairs of pyramidal neurons in both layer V (around 12%) and layer III (around 9%). In complete contrast, we have failed to find any recurrent synaptic connections between principal neurons in layer II, and this may be an important factor in the relative resistance of this layer in generating synchronized epileptiform activity. In general, recurrent excitatory postsynaptic potentials in layers III and V of the entorhinal cortex had similar properties to those recorded in other cortical areas, although the probabilities of connection are among the highest reported. Recurrent excitatory postsynaptic potentials recorded in layer V were smaller with faster rise times than those recorded in layer III. In both layers, the recurrent potentials were mediated by glutamate primarily acting at alpha amino-3-hydroxy-5-methyl-4-isoxazole receptors, although there appeared to be a slow component mediated by N-methyl-D-aspartate receptors. In layer III, recurrent transmission failed on about 30% of presynaptic action potentials evoked at 0.2Hz. This failure rate increased markedly with increasing (2, 3Hz) frequency of activation. In layer V the failure rate at low frequency was less (19%), and although it increased at higher frequencies this effect was less pronounced than in layer III. Finally, in layer III, there was evidence for a relatively high probability of electrical coupling between pyramidal neurons. We have previously suggested that layers IV/V of the entorhinal cortex readily generate synchronized epileptiform discharges, whereas layer II is relatively resistant to seizure generation. The present demonstration that recurrent excitatory connections are widespread in layer V but not layer II could support this proposal. The relatively high degree of recurrent connections and electrical coupling between layer III cells may be a factor in it's susceptibility to neurodegeneration during chronic epileptic conditions. PMID- 11029535 TI - Species-specific expression of parvalbumin in the entorhinal cortex of the Mongolian gerbil: dependence on local activity but not extrinsic afferents. AB - Mongolian gerbils are genetically predisposed to develop epileptic seizures in limbic structures. A species-specific property of the Mongolian gerbil is the expression of the calcium-binding protein parvalbumin in the perforant path where it is predominantly concentrated in nerve terminals. To test the hypothesis that this atypical expression of parvalbumin is induced by seizure-correlated hyperactivity in the entorhinohippocampal loop, we investigated whether it is dependent on extrinsic afferents to the entorhinal cortex. We cultivated organotypic slice cultures of neonate gerbil entorhinal cortex, isolated from all regions it is normally connected with in vivo. In these cultures, parvalbumin expressing neurons demonstrated their characteristic features like in vivo. Blockade of spontaneous local activity with the sodium-channel blocker tetrodotoxin, however, considerably reduced the number of parvalbumin-expressing neurons in culture. These results indicate that spontaneous local activity, but not activity mediated by extrinsic afferents, is an essential factor for the expression of parvalbumin in the entorhinal cortex of the Mongolian gerbil. PMID- 11029536 TI - Neuronal and behavioral correlations in the medial prefrontal cortex and nucleus accumbens during cocaine self-administration by rats. AB - Up to 31 neurons per animal were simultaneously recorded from the medial prefrontal cortex and nucleus accumbens in 15 rats during i.v. cocaine self administration sessions, using a multi-channel, single-unit recording technique. Alterations of neuronal activity (both excitatory and inhibitory) were found a few seconds before each lever press for cocaine infusion; we have called these pre-lever press neuronal activations "anticipatory responses". A detailed video analysis revealed that these neuronal firing alterations were associated with specific portions of the behavioral sequence performed before each lever press in both recording areas. Some of the simultaneously recorded neurons displayed similar firing patterns in relation to a given behavioral episode within the behavioral sequence (turning, raising head, etc.), while others fired at different times relative to each behavioral event. Cross-correlational analyses revealed inter-regional and intra-regional correlated firing patterns between pairs of simultaneously recorded medial prefrontal cortex and nucleus accumbens neurons. This correlated firing occurred in the neurons with and without anticipatory responses, although the incidence of correlations between anticipatory neuron pairs was much higher than that between non-anticipatory neuron pairs (18.4% vs 3.6%). Many correlated neuron pairs displayed a time lag in the peak of correlational activity that indicated a temporal sequence in correlated activity. In contradiction to our hypothesis, the temporal pattern of correlation reveals that there are more cases in which nucleus accumbens neurons fired ahead of medial prefrontal cortex neurons. The results suggest that multiple mesocorticolimbic neuronal circuits may code sequential steps during the behavioral sequence performed to obtain an infusion of cocaine. The observed correlated firing between the medial prefrontal cortex and the nucleus accumbens indicates that dynamic, coherent activity occurs within the mesocorticolimbic circuit. Because this circuit is hypothesized to drive drug-seeking behavior, we suggest that this correlated firing between the nucleus accumbens and the medial prefrontal cortex may participate in the control of cocaine self-administration. In addition, the finding that correlated activity within the nucleus accumbens more often precedes that of the medial prefrontal cortex suggests that the nucleus accumbens may play a prime role in the initiation of cocaine self administration. PMID- 11029537 TI - Mesoaccumbens dopamine neuron synapses reconstructed in vitro are glutamatergic. AB - The mesoaccumbens projection, formed by ventral tegmental area dopamine neurons synapsing on nucleus accumbens gamma-aminobutyric acid neurons, has been implicated in the pathogenesis of schizophrenia and drug addiction. Despite intensive study, the nature of the signal conveyed by dopamine neurons has not been fully resolved. In addition to several slower, dopamine-mediated, modulatory actions, several lines of evidence suggest that dopamine neurons have fast excitatory actions. To test this, we placed dopamine neurons together with accumbens neurons in microcultures. Surprisingly, most dopamine neurons made excitatory recurrent connections (autapses), which provided a basis for their identification; accumbens gamma-aminobutyric acid neurons were identified by their distinctive size. In 75% of mesoaccumbens cell pairs, stimulation of the dopamine neuron evoked a glutamate-mediated, excitatory synaptic response in the accumbens neuron. Immunostaining revealed dopamine neuron varicosities that were predominantly dopaminergic, ones that were predominantly glutamatergic, and ones that were both dopaminergic and glutamatergic. Despite close appositions of both glutamatergic and dopaminergic varicosities to the dendrites of accumbens neurons, only glutamatergic synaptic responses were seen. In the majority of cell pairs, pharmacologic activation of D2-type dopamine receptors inhibited glutamatergic responses, presumably via immunocytochemically-visualized presynaptic D2 receptors. In some cell pairs, the evoked autaptic and synaptic responses were discordant, suggesting that D2 receptors may be differentially trafficked to different presynaptic varicosities.Thus, dopamine neurons appear to mediate both slow dopaminergic and fast glutamatergic actions via separate sets of synapses. Together with evidence for glutamate cotransmission in serotonergic raphe neurons and noradrenergic locus coeruleus neurons, these results add a new dimension to monoamine neuron signaling that may have important implications for neuropsychiatric disorders. PMID- 11029538 TI - L-DOPA produces strong induction of c-fos messenger RNA in dopamine-denervated cortical and striatal areas of the common marmoset. AB - Common marmosets (Callithrix jacchus) with near-complete unilateral 6 hydroxydopamine denervation of the dopaminergic input received a single injection of saline or L-DOPA (15mg/kg plus 6.25mg/kg benserazide). Using in situ hybridization, the effects of these treatments on c-fos messenger RNA expression in the cerebral cortex, the striatal complex and the external layer of the pallidum were studied. Moreover, receptor autoradiography was used to determine the levels of dopamine D(1) and D(2) receptors in these areas. In the cerebral cortex, animals treated with L-DOPA displayed a high expression of c-fos messenger RNA restricted to the dopamine-denervated hemisphere. No changes in the levels of cortical D(1) and D(2) receptors were found in the dopamine-denervated hemisphere. L-DOPA treatment also induced a strong expression of c-fos messenger RNA in the striatal complex in the dopamine-denervated hemisphere. The levels of striatal D(2), but not D(1), receptors were increased in the dopamine-denervated hemisphere. In the external pallidum, the major terminal region for D(2) dopamine receptor-containing striatal projection neurons, L-DOPA treatment induced c-fos messenger RNA expression in both the intact and the dopamine-denervated hemispheres.Thus, using c-fos messenger RNA as a biochemical marker of postsynaptic neuronal activation, these results provide evidence that near complete dopamine depletion causes a profound supersensitization to L-DOPA treatment in the cerebral cortex and in the striatal complex, but not in the external layer of the pallidum, of the primate brain. The cortical response may be unique to the primate brain, but c-fos messenger RNA activation within the striatum has also been reported in the rodent. The effects of L-DOPA probably depend both on a direct activation of supersensitized dopamine receptors by dopamine produced in the few remaining, but hyperactive, dopaminergic nerve terminals and in serotonergic nerve terminals, as well as on indirect actions of L-DOPA related to activation of circuitries connecting cerebral cortex and basal ganglia structures. These results provide novel information on the mechanisms underlying L-DOPA's action in the cerebral cortex, striatum and external pallidum in a primate model of Parkinson's disease. PMID- 11029539 TI - The lesional and epileptogenic consequences of lithium-pilocarpine-induced status epilepticus are affected by previous exposure to isolated seizures: effects of amygdala kindling and maximal electroshocks. AB - In temporal lobe epilepsy, the occurrence of seizures seems to correlate with the presence of lesions underlying the establishment of a hyperexcitable circuit. However, in the lithium-pilocarpine model of epilepsy, neuronal damage occurs both in the structures belonging to the circuit of initiation and maintenance of the seizures (forebrain limbic system) as in the propagation areas (cortex and thalamus) and in the circuit of remote control of seizures (substantia nigra pars reticulata). To determine whether or not we could protect the brain from lesions and epileptogenesis induced by status epilepticus and identify cerebral structures involved in the genesis of epilepsy, we studied the effects of the chronic exposure to non-deleterious seizures, either focalized with secondary generalization (amygdala kindling, kindled-pilocarpine rats), or primary generalized (ear-clip electroshocks, electroshock-pilocarpine rats) on neuronal damage and epileptogenesis induced by lithium-pilocarpine status epilepticus. These animals were compared to rats subjected to status epilepticus but not pretreated with seizures (sham-kindled-pilocarpine or sham-electroshock pilocarpine rats). Compared to sham-pilocarpine rats, neuronal damage was prevented in the limbic system of the kindled-pilocarpine rats, except in the hilus of the dentate gyrus and the entorhinal cortex, while it was enhanced in rats pretreated with electroshocks, mainly in the entorhinal and perirhinal cortices. Most sham-kindled- and sham-electroshock-pilocarpine rats (92-100%) developed recurrent seizures after a silent period of 40-54days. Likewise, all kindled-pilocarpine rats developed spontaneous seizures after the same latency as their sham controls, while only two of 10 electroshock-pilocarpine rats became epileptic after a delay of 106-151days. The present data show that the apparent antiepileptic properties of electroshocks correlate with extensive damage in midbrain cortical regions, which may prevent the propagation of seizures from the hippocampus and inhibit their motor expression. Conversely, the extensive neuroprotection of the limbic system but not the hilus and entorhinal cortex provided by amygdala kindling does not prevent epileptogenesis. Thus, the hilus, the entorhinal and/or perirhinal cortex may be key structure(s) for the establishment of epilepsy. PMID- 11029540 TI - Brain-derived neurotrophic factor administration after traumatic brain injury in the rat does not protect against behavioral or histological deficits. AB - Brain-derived neurotrophic factor has been shown to be neuroprotective in models of excitotoxicity, axotomy and cerebral ischemia. The present study evaluated the therapeutic potential of brain-derived neurotrophic factor following traumatic brain injury in the rat. Male Sprague-Dawley rats (N=99) were anesthetized and subjected to lateral fluid percussion brain injury of moderate severity (2.4-2.8 atm) or sham injury. Four hours after injury, the animals were reanesthetized, an indwelling, intraparenchymal cannula was implanted, and infusion of brain-derived neurotrophic factor or phosphate-buffered saline vehicle was initiated from a mini-osmotic pump and continued for two weeks. In Study 1 (N=48), vehicle or 12 microg/day of brain-derived neurotrophic factor was infused into the dorsal hippocampus. In Study 2 (N=51), vehicle or brain-derived neurotrophic factor at a high (12 microg/day) or low dose (1.2 microg/day) was infused into the injured parietal cortex. All animals were evaluated for neurological motor function at two days, one week and two weeks post-injury. Cognitive function (learning and memory) was assessed at two weeks post-injury using a Morris Water Maze. At two weeks post-injury, neuronal loss in the hippocampal CA3 and dentate hilus and in the injured cortex was evaluated. In Study 2, neuronal loss was also quantified in the thalamic medial geniculate nucleus. All of the above outcome measures demonstrated significant deleterious effects of brain injury (P<0.05 compared to sham). However, post-traumatic brain-derived neurotrophic factor infusion did not significantly affect neuromotor function, learning, memory or neuronal loss in the hippocampus, cortex or thalamus when compared to vehicle infusion in brain injured animals, regardless of the infusion site or infusion dose (P>0.05 for each). In contrast to previous studies of axotomy, ischemia and excitotoxicity, our data indicate that brain-derived neurotrophic factor is not protective against behavioral or histological deficits caused by experimental traumatic brain injury using the delayed, post-traumatic infusion protocol examined in these studies. PMID- 11029541 TI - Synergistic sedative effects of noradrenergic alpha(1)- and beta-receptor blockade on forebrain electroencephalographic and behavioral indices. AB - The locus coeruleus-noradrenergic system exerts an activating influence on forebrain neuronal and behavioral activity states. For example, in the anesthetized rat, unilateral locus coeruleus stimulation elicits bilateral activation of forebrain electroencephalographic activity. Pretreatment with a noradrenergic beta-antagonist blocks this effect, suggesting that beta-receptors play a critical role in locus coeruleus-dependent activation of the forebrain. Consistent with this, stimulation of beta-receptors located in certain basal forebrain structures evokes sustained periods of alert waking in the unanesthetized rat. Similar forebrain and behavioral activating effects are observed with alpha(1)-receptor stimulation within these basal forebrain regions. To assess the extent to which alpha(1)- and beta-receptors contribute to the maintenance of behavioral and forebrain activation, we examined the electroencephalographic and behavioral effects of alpha(1)-, beta- and combined alpha(1)/beta-receptor blockade in the unanesthetized rat. Rats were treated individually or in combination with either varying doses of the alpha(1) antagonist, prazosin (intraperitoneally), and/or the beta-antagonist, timolol (intracerebroventricularly). Thirty minutes following treatment, animals were placed in a mildly-arousing novel environment, which has been demonstrated previously to elicit activation of central noradrenergic systems and sustained waking in vehicle-treated controls. Behavior and electroencephalographic activity were recorded and later scored. Electroencephalographic activity was analysed using power spectrum analysis. The following were observed: (i) beta-receptor blockade alone does not alter behavioral or electroencephalographic indices of alert waking; (ii) alpha(1)-receptor blockade alone increases high-voltage spindle activity in cortical electroencephalographic activity that was associated with decreased behavioral activity; (iii) combined alpha(1)- and beta-receptor blockade elicits a substantial increase in slow-wave activity (0.33-2.0Hz), also in association with decreased behavioral activity. All of these effects were dependent on the dose administered and time following initiation of testing. These results indicate that the combined actions of alpha(1)- and beta-receptors exert distinct and synergistic actions on cortical neuronal activity patterns that are essential elements of alert waking. PMID- 11029543 TI - Serotonin uptake and release mechanisms in developing cultures of rat embryonic raphe neurons: age- and region-specific differences. AB - The development of serotonergic neurons of the rat raphe was followed in primary neuronal cell cultures taken at embryonic days embryonic day 13 and embryonic day 14 from three different raphe sub-groups, topographically defined with respect to their position to the isthmus as rostral (R1), intermediate (R2) and caudal (R3). In neurons cultivated from embryonic day 13 raphe serotonin, immunoreactivity was detected after only two days in vitro in the rostral R1 and the intermediate R2 sub-groups. Within two weeks of cultivation the number of serotonergic neurons as well as the dendritic branching continuously increased in all three sub-groups. In cultures obtained from embryonic day 13 raphe a specific uptake of [3H]serotonin could not be detected during the first days in vitro. Specific uptake as well as regulated serotonin release, however, was clearly discernible in these cultures after nine days in vitro, indicating developmental differentiation of the initially immature serotonergic neurons in culture. In contrast, serotonergic neurons obtained from the three raphe sub-groups at embryonic day 14 took up and released [3H]serotonin, as early as after two days in culture. Basal as well as stimulated serotonin release was diminished when preincubating the cells with tetanus toxin, which cleaves synaptobrevin thereby blocking exocytosis. Our data indicate that the differential development of serotonergic neurons in the various raphe sub-groups in vivo is also sustained in culture. The differences observed when comparing neurons from embryonic days 13 and 14 suggest that a short time-period of about 24h appears to be crucial for the developmental upregulation of serotonin uptake, storage and release. PMID- 11029542 TI - Brain site-specificity of extracellular adenosine concentration changes during sleep deprivation and spontaneous sleep: an in vivo microdialysis study. AB - Previous data suggested that increases in extracellular adenosine in the basal forebrain mediated the sleep-inducing effects of prolonged wakefulness. The present study sought to determine if the state-related changes found in basal forebrain adenosine levels occurred uniformly throughout the brain. In vivo microdialysis sample collection coupled to microbore high-performance liquid chromatography measured extracellular adenosine levels in six brain regions of the cat: basal forebrain, cerebral cortex, thalamus, preoptic area of hypothalamus, dorsal raphe nucleus and pedunculopontine tegmental nucleus. In all these brain regions extracellular adenosine levels showed a similar decline of 15 to 20% during episodes of spontaneous sleep relative to wakefulness. Adenosine levels during non-rapid eye movement sleep did not differ from rapid eye movement sleep. In the course of 6h of sleep deprivation, adenosine levels increased significantly in the cholinergic region of the basal forebrain (to 140% of baseline) and, to a lesser extent in the cortex, but not in the other regions. Following sleep deprivation, basal forebrain adenosine levels declined very slowly, remaining significantly elevated throughout a 3-h period of recovery sleep, but elsewhere levels were either similar to, or lower than, baseline. The site-specific accumulation of adenosine during sleep deprivation suggests a differential regulation of adenosine levels by as yet unidentified mechanisms. Moreover, the unique pattern of sleep-related changes in basal forebrain adenosine level lends strong support to the hypothesis that the sleep-promoting effects of adenosine, as well as the sleepiness associated with prolonged wakefulness, are both mediated by adenosinergic inhibition of a cortically projecting basal forebrain arousal system. PMID- 11029544 TI - Morphine treatment induced calcitonin gene-related peptide and substance P increases in cultured dorsal root ganglion neurons. AB - The mechanism of spinal tolerance to the analgesic effects of opiates is unclear at present. We have reported previously that calcitonin gene-related peptide-like immunoreactivity was significantly increased in primary afferents of the spinal dorsal horn during the development of morphine tolerance, suggesting that changes in the level of pain-related neuropeptides in dorsal root ganglion neurons may be involved [Menard D. P. et al. (1996) J. Neurosci. 16, 2342-2351]. In this study, we investigated if in vitro treatment with morphine can mimic the in vivo findings and induce increases in calcitonin gene-related peptide-like immunostaining in cultured dorsal root ganglion neurons from young (three-month old) and middle-aged (10-month-old) adult rats. Following a repetitive exposure to morphine sulfate (1, 5, 10 microM) for six days, the number of calcitonin gene related peptide- and substance P-immunoreactive neurons in cultured dorsal root ganglia from three- and 10-month-old rats was significantly increased. A lower concentration (0.5 microM) of morphine induced these increases only in dorsal root ganglion neurons from middle-aged rats. Morphine treatment was also found to increase the number of calcitonin gene-related peptide-immunoreactive neurons possessing multiple, long branches (i.e. with at least one branch >0.5mm). This apparent increase in the number of calcitonin gene-related peptide- and substance P-immunoreactive neurons observed following morphine treatment was blocked by naloxone, an opiate antagonist, indicating the involvement of genuine opioid receptors. No significant change in the number of neuropeptide Y- or galanin immunoreactive neurons in cultured dorsal root ganglia was detected following any of these treatments. These data suggest that repeated exposure to morphine rather selectively increases calcitonin gene-related peptide- and substance P-like immunoreactivity in cultured dorsal root ganglion neurons. Moreover, the sensitivity to morphine-induced changes is greater in cultured dorsal root ganglion neurons from 10- compared to three-month-old rats. Hence, cultured dorsal root ganglion neurons can provide a model to investigate the cellular and molecular mechanisms underlying alterations in neuropeptide levels following repeated exposure to opiates and their relevance to the development of opioid tolerance. PMID- 11029545 TI - Inhibitory effects evoked from the rostral ventrolateral medulla are selective for the nociceptive responses of spinal dorsal horn neurons. AB - The aim of the present study was to determine whether or not descending control of spinal dorsal horn neuronal responsiveness following neuronal activation at pressor sites in the rostral ventrolateral medulla is selective for nociceptive information. Extracellular single-unit activity was recorded from 49 dorsal horn neurons in the lower lumbar spinal cord of anaesthetized rats. The 30 Class 2 neurons selected for investigation responded to noxious (pinch and radiant heat) and non-noxious (prod, stroke and/or brush) stimulation within their cutaneous receptive fields on the ipsilateral hindpaw. The excitatory amino acid, DL homocysteic acid, was microinjected into either the rostral or the caudal rostral ventrolateral medulla at sites that evoked increases in arterial blood pressure. Effects of neuronal activation at these sites were then tested on the responses of Class 2 neurons to noxious and non-noxious stimulation within their excitatory receptive fields. The noxious pinch and radiant heat responses of Class 2 neurons were depressed, respectively to 13+/-3.8% (n=23) and to 16+/-3.7% (n=18) of control, following stimulation at sites in the rostral rostral ventrolateral medulla. In contrast, the low-threshold (prod) responses of eight Class 2 neurons tested were not depressed following neuronal activation at the same sites. When tested, control injections of the inhibitory amino acid, GABA, at the same sites in the rostral rostral ventrolateral medulla had no significant effects on neuronal activity. Neither intravenous administration of noradrenaline (to mimic the pressor responses evoked by DL-homocysteic acid microinjections in the rostral ventrolateral medulla) nor activation at pressor sites in the caudal rostral ventrolateral medulla had any significant effect on neuronal responsiveness. With regard to sensory processing in the spinal cord, these data suggest that descending inhibitory control that originates from neurons in pressor regions of the rostral rostral ventrolateral medulla is highly selective for nociceptive inputs to Class 2 neurons. These data are discussed in relation to the role of the rostral ventrolateral medulla in executing the changes in autonomic and sensory functions that are co-ordinated by higher centres in the CNS. PMID- 11029546 TI - Alteration in synaptic inputs through C-afferent fibers to substantia gelatinosa neurons of the rat spinal dorsal horn during postnatal development. AB - The change in synaptic inputs through primary afferent C- and A-fibers during postnatal development was examined in substantia gelatinosa neurons of a rat spinal cord slice with an attached L5 dorsal root by use of the blind whole-cell patch-clamp technique; the synaptic responses were compared between the slices obtained from immature (postnatal days 21-23) and mature (postnatal days 56-60) male rats. The mono- and/or polysynaptic afferent inputs were monitored by recording glutamatergic excitatory postsynaptic currents and potentials evoked by stimulating C- and A-fibers, the identification of which was based on the values of threshold stimulus intensity and of the conduction velocity of the fibers, determined by intracellular recordings from dorsal root ganglion neurons. Immature substantia gelatinosa neurons received synaptic inputs through Abeta-, Adelta- and C-afferents, with proportions of 51%, 46% and 36%, respectively. In mature substantia gelatinosa neurons, C- and Adelta-afferent inputs were increased in number (to 84% and 86%, respectively), while Abeta-inputs were decreased to 9%. In both immature and mature rats, repetitive stimulation of C afferents did not elicit any slow responses, which are longer in duration than the monosynaptic excitatory postsynaptic currents, although C-fibers are known to contain not only excitatory amino acids, but also neuropeptides such as substance P, which is thought to be involved in the production of slow responses. These results indicate that both C- and Adelta-afferents innervating substantia gelatinosa neurons are reorganized following maturation, accompanied by a withdrawal or elimination of Abeta-fibers from the substantia gelatinosa, probably due to a competition among the fibers during development. In spite of the developmental increase in C-fiber inputs, mature as well as immature substantia gelatinosa neurons did not display any slow synaptic responses, which appear to be mediated by transmitters other than excitatory amino acids. PMID- 11029547 TI - Cellular and subcellular distribution of the amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor subunit GluR2 in the rat dorsal vagal complex. AB - Amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) type glutamate receptors are ligand gated ion channels made up of various combinations of four subunits termed GluR1-4. The GluR2 subunit controls several key features of the receptor including calcium permeability and inward rectification. In the present study, we analysed by immunocytochemistry the cellular and subcellular distribution of the GluR2 subunit in neurons of the dorsal vagal complex of the rat. GluR2 immunoreactivity was found both in the neuropile and in neuronal cell bodies. Perikaryal staining was strong in the dorsal motor nucleus of the vagus nerve and moderate in the medial part of the nucleus tractus solitarii as well as in the area postrema. The lateral part of the nucleus tractus solitarii was almost devoid of immunoreactivity except for the interstitial subnucleus which was filled with numerous strongly immunoreactive perikarya and large cell processes. Ultrastructural examination was carried out in the interstitial subnucleus. Peroxidase staining indicative of GluR2 immunoreactivity was observed in neuronal cell bodies and dendrites. No labeled axon terminal or glial cell body was found. Additional experiments performed using pre-embedding immunogold showed that most of the labeling in immunoreactive dendrites was intracytoplasmic. These results indicate that GluR2 immunoreactivity is differentially distributed among neurons in the dorsal vagal complex, thereby suggesting differences in the functional properties of AMPA receptors between neuronal populations. These results also suggest that AMPA receptors, at least those containing the GluR2 subunit, have no major role as presynaptic receptors within this region. Finally, they indicate the existence of large intracellular pools of GluR2 subunits within dendrites of immunoreactive neurons. PMID- 11029548 TI - Calbindin and Fos within the suprachiasmatic nucleus and the adjacent hypothalamus of Arvicanthis niloticus and Rattus norvegicus. AB - The suprachiasmatic nucleus is the site of the primary circadian pacemaker in mammals. The lower sub paraventricular zone that is dorsal to and receives input from the suprachiasmatic nucleus may also play a role in the regulation of circadian rhythms. Calbindin has been described in the suprachiasmatic nucleus of some mammals, and may be important in the control of endogenous rhythms. In the first study we characterized calbindin-expressing cells in the suprachiasmatic nucleus and lower sub-paraventricular zone of nocturnal and diurnal rodents. Specifically, Rattus norvegicus was compared to Arvicanthis niloticus, a primarily diurnal species within which some individuals exhibit nocturnal patterns of wheel running. Calbindin-immunoreactive cells were present in the suprachiasmatic nucleus of Arvicanthis and were most concentrated within its central region but were relatively sparse in the suprachiasmatic nucleus of Rattus. Calbindin-expressing cells were present in the lower sub-paraventricular zone of both species. In the second study we evaluated Fos expression within calbindin-immunoreactive cells in nocturnal Rattus and in Arvicanthis that were either diurnal or nocturnal with respect to wheel-running. All animals were kept on a 12:12 light/dark cycle and perfused at either 4h after lights-on or 4h after lights-off. In the suprachiasmatic nucleus in both species, Fos expression was elevated during the day relative to the night but less than 1% of calbindin cells contained Fos in Arvicanthis, compared with 13-17% in Rattus. In the lower sub paraventricular zone of both species, 9-14% of calbindin cells expressed Fos, and this proportion did not change as a function of time. Among Arvicanthis, the number of calbindin expressing neurons in the lower sub-paraventricular zone was influenced by an interaction between the wheel running patterns (nocturnal vs diurnal) and time of day. Thus, the number of calbindin-positive cells within the suprachiasmatic nucleus differed in Arvicanthis and Rattus, whereas the number of calbindin-positive cells within the lower sub-paraventricular zone differed in nocturnal and diurnal Arvicanthis. Our examination of R. norvegicus and A. niloticus suggests potentially important relationships between calbindin containing neurons and whether animals are nocturnal or diurnal. Specifically, rats had more Fos expression in calbindin containing cells in the suprachiasmatic nucleus than Arvicanthis. In contrast, Arvicanthis exhibiting diurnal and nocturnal patterns of wheel-running differed in the number of calbindin containing cells in the lower sub-paraventricular zone, dorsal to the suprachiasmatic nucleus. PMID- 11029549 TI - Induction of vascular endothelial growth factor and hypoxia-inducible factor 1alpha by global ischemia in rat brain. AB - Vascular endothelial growth factor is an angiogenic and neurotrophic peptide whose expression is transcriptionally induced in hypoxic tissues through the action of hypoxia-inducible factor-1alpha. To determine if this signaling pathway is activated in the ischemic brain, and might therefore participate in adaptive processes such as angiogenesis and neuroprotection, we examined the expression of vascular endothelial growth factor and hypoxia-inducible factor-1alpha in cerebral cortex and hippocampus following transient global cerebral ischemia in the rat. Northern analysis showed ischemia-inducible expression of multiple vascular endothelial growth factor messenger ribonucleic acid splice variants between 4 and 24h. Western analysis and immunocytochemistry demonstrated the concerted induction of vascular endothelial growth factor and hypoxia-inducible factor-1alpha in the same, apparently neuronal, cells in vulnerable regions of cortex and hippocampus after 15min of ischemia, which persisted for as long as 4 to 72h of reperfusion. These findings demonstrate that hypoxia-sensitive vascular endothelial growth factor signaling can be induced in neurons in global cerebral ischemia in vivo, and are consistent with the hypothesis that ischemic insults trigger hypoxia-sensing and adaptive downstream molecular responses in central neurons. PMID- 11029550 TI - Towards a closed eye model of the pre-ocular tear layer. AB - Although the tear film has been extensively studied as it exists in the open eye state, until recently very little was known as to what happens to the tear film on eye closure. Recent studies have shown that eye closure results in a profound change in the composition, origins, turnover and physiological functions of the tear film. These changes include a shift from an inducible, neurologically controlled, lacrimal secretion containing among other proteins primarily lysozyme, lactoferrin and tear specific lipocalin, to a much slower, constitutive type of secretion, composed almost exclusively of sIgA. This change is accompanied by the build-up of sialoglycoproteins of epithelial and goblet cell origin, the build-up and activation of complement and the build-up of serum proteins. In addition, various cytokines and proinflammatory mediators accumulate, including some which are potent inducers of angiogenesis and leukochemotaxis. The closed eye also exhibits the recruitment and activation of massive numbers of PMN cells. This results in a stagnant, closed eye layer, which is extremely rich in reactive complement products, PMN cell proteases including protease-3, elastase, capthepsin G, MMP-9 and urokinase. We have postulated that this shift represents a fundamental change in host-defense strategies from a passive-barrier defense to an active immune, inflammatory, phagocyte-mediated process and that this shift is necessitated in order to protect the cornea from entrapped microorganisms. Studies have shown that autologous cell damage is avoided in closed eye tear fluid, by the accumulation of several modulators of complement activation, which shift activation towards opsonization of entrapped microorganisms and the build-up of a wide array of antiproteases. Some of the latter are likely to arise from the ocular surface tissues. Corneal neovascularization may be avoided in part by the build-up of alpha2-macroglobulin and the conversion of plasminogen to angiostatin. It is highly probable that other bioactive protein fragments are produced in the closed eye, which contribute to homeostasis. Areas of future study are indicated. PMID- 11029551 TI - VIP enhances the differentiation of retinal pigment epithelium in culture: from cAMP and pp60(c-src) to melanogenesis and development of fluid transport capacity. AB - The retinal pigment epithelium (RPE) is a single cell layer juxtaposed between the neural retina and the choroid and functions as a blood-retina barrier. The RPE performs functions essential for photoreceptor (PR) survival. Although the regulation of these functions has remained unknown, it is a distinct possibility that the RPE is under constant regulation by signaling molecules coming from the choroid and the retina. Vasoactive intestinal peptide (VIP), a 28-amino acid neuropeptide present in the retina and in the choroid, has been shown to promote the growth and differentiation of a variety of cells in tissue and organ cultures. In cultured RPE cells, VIP is the one most effective stimulator of the cAMP signaling pathway among a long list of neurotransmitters and modulators tested. For example, VIP, at 1 microM, stimulates the intracellular cAMP to 80 100- and 20-fold in 3 min in RPE cells cultured from chick embryos and adult human donor eyes, respectively. In cultured chick embryonic RPE, VIP is also shown to be a potent and effective modulator of pp60(c-src), the non-receptor tyrosine kinase present in differentiating and terminally differentiated cells. VIP stimulates both overall phosphorylation at unknown sites and phosphotyrosine dephosphorylation in pp60(c-src). A 190-kDa microtubule-associated protein is known to be one of the downstream targets in VIP-modulated signaling pathways. At the cellular level, VIP stimulates cell proliferation modestly and melanogenesis pronouncedly in growing chick embryonic RPE cultures. Ultimately, the differentiation goal of RPE cells in vivo is to perform functions that are essential for photoreceptor survival. On bare permeable supports (that is, without biological material coating), the chick embryonic RPE cells grow to become RPE sheets with a cytoarchitecture that allows the display of two of the RPE functions. These cultures demonstrate structural polarity and are functionally polarized, allowing for proper macromolecule secretion and fluid transport. VIP is shown to stimulate macromolecule secretion at the apical surface (retina facing) and the development of the capacity for fluid transport from the apical to the basal surface of the RPE sheet. In conclusion, studies in our laboratory indicate that VIP is a differentiation promotor during the development of a functional RPE. Recent advances in the molecular biology of melanogenesis and the fluid transport-linked Na-K-2Cl cotransporter in other cells will allow future studies of VIP modulated events in the RPE at the molecular level. Finally, identification of RPE differentiation factors may prove essential for the ultimate success of RPE transplantation, thus promoting the rescue of photoreceptor cells in retinal degeneration. PMID- 11029552 TI - Ribozyme uses in retinal gene therapy. AB - In this chapter we discuss the design, delivery and preclinical testing of mutation-specific ribozymes for the treatment of dominantly inherited retinal disease. We focus particular attention on the initial screening of ribozymes in vitro, because the activity of RNA enzymes in cell-free systems can be used to predict their suitability for animal experiments. Current techniques for delivering genes of interest to cells of the retina using viral vectors are then briefly surveyed emphasizing vector properties that best match to the needs of a ribozyme-based therapy. Using these considerations, analysis of ribozyme gene therapy for an autosomal dominant RP-like disease in a rodent model is outlined emphasizing the desirability of combining biochemical, morphological and electrophysiological measures of therapy. Finally, we describe alternative, perhaps more general, ribozyme approaches that have yet to be tested in the context of retinal disease. PMID- 11029553 TI - The mammalian photoreceptor mosaic-adaptive design. AB - Unlike in birds and cold-blooded vertebrates' retinas, the photoreceptors of mammalian retinas were long supposed to be morphologically uniform and difficult to distinguish into subtypes. A number of new techniques have now begun to overcome the previous limitations. A hitherto unexpected variability of spectral and morphological subtypes and topographic patterns of distribution in the various retinas are being revealed. We begin to understand the design of the photoreceptor mosaics, the constraints of evolutionary history and the ecological specialization of these mosaics in all the mammalian subgroups. The review discusses current cytological identification of mammalian photoreceptor types and speculates on the likely "bottleneck-scenario" for the origin of the basic design of the mammalian retina. It then provides a brief synopsis of current data on the photoreceptors in the various mammalian orders and derives some trends for phenomena such as rod/cone dualism, spectral range, preservation or loss of double cones and oil droplets, photopigment co-expression and mono- and tri chromacy. Finally, we attempt to demonstrate that, building on the limits of an ancient rod dominant (probably dichromatic) model, mammalian retinas have developed considerable radiation. Comparing the nonprimate models with the intensively studied primate model should provide us with a deeper understanding of the basic design of the mammalian retina. PMID- 11029554 TI - Hepatocyte growth factor/scatter factor in the eye. AB - Hepatocyte growth factor, also known as scatter factor (HGF/SF) is a multipotential cytokine which can produce a range of responses in target cells and its influence in the eye in health and disease is just beginning to be appreciated. Usually HGF/SF is synthesised by mesenchymally derived cells and targets and signals epithelial cells in a paracrine manner via their c-Met surface receptor. However, there is growing evidence for the existence of autocrine loops in a number of cell systems prominent among which are ocular cells such as the corneal endothelium, the lens epithelium, the retinal pigment epithelium (RPE) and others. Marked cellular proliferation is stimulated when activated HGF/SF is exposed to hepatocytes, renal epithelium, melanocytes and vascular endothelial cells but it is often a poor mitogen for other cell types. In target cells the cytokine promotes other bioactions such as junctional breakdown, shape change, cell scattering, directional and nondirectional migration, cell survival, invasive behaviour and/or tubule formation. These activities seem to depend on HGF/SF linking with the c-Met receptor and pathways to stimulate the various types of cytokine/receptor response are being unravelled at the present time. In corneal wound healing, HGF/SF is produced by stromal keratocytes and targets the repairing epithelium. HGF/SF is a constituent of tears, aqueous humour and vitreous humour at levels above that found in plasma although it is not clear how much is activated. Aqueous HGF/SF may well influence lens epithelial, corneal endothelial and trabecular meshwork cell survival. Vitreous levels of HGF/SF are elevated in proliferative vitreoretinopathy (PVR), where a target cell is the RPE and in proliferative diabetic retinopathy (PDR) where HGF/SF has been shown to be a major angiogenesis factor. Finally HGF/SF may be involved in the metastatic spread of tumour cells from uveal melanomata and in the formation of vascular channels in these tumours. PMID- 11029555 TI - Viscosupplementation: therapeutic mechanisms and clinical potential in osteoarthritis of the knee. AB - Viscosupplementation by means of intra-articular injections of hyaluronic acid has been used to treat osteoarthritis of the knee. The proposed mechanisms of action result from the physical properties of hyaluronic acid, as well as from its anti-inflammatory, anabolic, local analgesic, and chrondroprotective effects. Adverse reactions from hyaluronic acid injections into the knee occurred in 8.3% of the 336 patients treated in one study, but at a rate of less than 3% per injection. Reactions were almost always local and generally resolved over 1 to 2 days. Hyaluronic acid injections were approved by the US Food and Drug Administration as a medical device; thus, the level of efficacy demonstrated is less than might have been required for approval as a drug. Several studies have failed to show statistically significant benefit compared with placebo. Furthermore, the treatment is relatively expensive; the cost of the drug for a series of injections is more than $500 per knee. Therefore, widespread use of these agents should be limited until more convincing data on their efficacy are available from well-designed clinical trials. PMID- 11029556 TI - Pathophysiology of infections after internal fixation of fractures. AB - Infection complicating internal fixation of fractures is a serious complication that is difficult to treat. Whenever metallic devices are implanted in vivo, successful biointegration requires that host cells colonize the highly reactive implant surface. Bacteria such as staphylococci can also become adherent to metallic or polymeric implants and will compete with host cells for colonization of the implant surface. Once adherent, these bacteria form a biofilm and undergo phenotypic changes that make them resistant to the normal host immune response as well as to antibiotics. Furthermore, metallic implants themselves cause specific deficits in the function of the local immune system that may render the host response to infection inadequate. Any associated soft-tissue injury causes even greater impairment of local immune function. Despite the potentially detrimental impact of internal fixation, fracture stability is of paramount importance in achieving fracture union and in preventing infection. It has been demonstrated in animal models that contaminated fractures without internal fixation develop clinical infection more commonly than similar fractures treated with internal fixation at the time of colonization. Because of the potential for infection whenever internal fixation is utilized, appropriate prophylactic antibiotic coverage for staphylococci and Gram-negative organisms should be provided. Open wounds and severely damaged soft tissues require aggressive management so that a viable soft-tissue envelope is maintained around the implant. Host factors such as smoking and malnourishment should be corrected. Early diagnosis and aggressive treatment of implant-related infection with antibiotics, debridement, and maintenance of stable internal fixation are essential to successful treatment. PMID- 11029557 TI - Cartilage tumors: evaluation and treatment. AB - The proper treatment of cartilaginous tumors is dependent on the clinicopathologic and radiologic findings. Enchondroma is a benign tumor that is usually asymptomatic and thus should be treated nonoperatively. Symptomatic enchondromas are often treated by intralesional excision. Intramedullary low grade chondrosarcoma is a malignant tumor that is usually painful. The treatment of low-grade chondrosarcoma may range from intralesional excision with or without adjuvant therapy to wide excision. Although intralesional excisions have a higher bone and joint preservation rate than wide excisions, they may be associated with a higher local recurrence rate. Intermediate- and high-grade chondrosarcomas are treated with wide excisions. The treatment of these cartilaginous lesions should involve a multidisciplinary team including a musculoskeletal surgeon, a radiologist, and a pathologist. PMID- 11029558 TI - Thermal modification of connective tissues: basic science considerations and clinical implications. AB - Thermal modification (shrinkage) of capsular connective tissue has gained increasing popularity as an adjunctive or even a primary procedure in the arthroscopic treatment of shoulder instability. Although the physical effects of heat on collagenous tissues are well known, the long-term biologic fate of these shrunken tissues is still a matter of debate. The temperatures required to alter the molecular bonding of collagen and thus cause tissue shrinkage (65 degrees C to 70 degrees C) are also known to destroy cellular viability. Therefore, thermally modified tissues are devitalized and must undergo a biologic remodeling process. During this remodeling, the mechanical properties of the treated tissues are altered (decreased stiffness) and can be at risk for elongation if the postoperative rehabilitation regimen is too aggressive. Although anecdotal reports suggest that thermal capsular shrinkage does have a beneficial effect, the exact mechanism responsible for this clinical improvement has yet to be fully defined. The reported improvement could be due to the maintenance of initial capsular shrinkage, secondary fibroplasia and resultant thickening of the joint capsule, a loss of afferent sensory stimulation due to the destruction of sensory receptors, or a combination of all three. The clinical role for thermal modification of connective tissues has not yet been defined, but it appears that it may prove most useful as a stimulant for inducing a biologic repair response. PMID- 11029559 TI - Basal joint arthritis of the thumb. AB - Thumb pain secondary to arthritis at the basal joint of the thumb is a common condition, especially in women, and can be quite disabling. An accurate diagnosis can be readily made from the history and examination. Radiographs are used to stage the severity of the arthritis. Splinting is the mainstay of conservative care. Reconstructive procedures for each stage of the disease are aimed at restoring thumb motion and strength. Partial or complete trapeziectomy with tendon interposition and ligament reconstruction to stabilize the metacarpal base is used for advanced disease. Secondary metacarpophalangeal joint hyper-extension deformity may need to be addressed. Surgery can reliably improve function and engender high patient satisfaction. PMID- 11029560 TI - Use of the pedicled latissimus muscle flap for upper-extremity reconstruction. AB - Tissue with a blood supply derived from a single constant vascular pedicle may be raised as a flap and rotated within the reach of its blood supply to cover and reconstruct a variety of complex wounds. The latissimus dorsi muscle makes an ideal pedicled flap because of its long neurovascular pedicle, large size, ease of mobilization, and expendability. It can be rotated, with or without overlying skin, to cover soft-tissue defects involving the shoulder, arm, and elbow, or it can be transferred as an innervated muscle to improve shoulder abduction as well as elbow flexion and extension. The major clinical applications of the pedicled latissimus dorsi muscle flap for upper-extremity reconstruction include use as a bipolar transfer to improve elbow flexion after trauma or brachial plexus injury and as a nonfunctioning myocutaneous transfer for coverage of nerves, bones, and joints after soft-tissue loss due to trauma, tumors, infection, or irradiation. PMID- 11029561 TI - Treatment strategies for acute fractures and nonunions of the proximal fifth metatarsal. AB - There are at least three distinct fracture patterns that occur in the proximal fifth metatarsal: tuberosity avulsion fractures, acute Jones fractures, and diaphyseal stress fractures. Each of these fracture patterns has its own mechanism of injury, location, treatment options, and prognosis regarding delayed union and nonunion. Tuberosity avulsion fractures are the most common in this region of the foot. The majority heal with symptomatic care in a hard-soled shoe. The true Jones fracture is an acute injury involving the fourth-fifth intermetatarsal facet. These injuries are best treated with non-weight-bearing cast immobilization for 6 to 8 weeks. The rate of successful union with this treatment has been reported to be between 72% and 93%. For the high-performance athlete with an acute Jones fracture, early intramedullary-screw fixation is an accepted treatment option. Nonacute diaphyseal stress fractures of the proximal fifth metatarsal and Jones fractures that develop into delayed unions and nonunions can both be managed with operative fixation with either closed axial intramedullary-screw fixation or autogenous corticocancellous grafting. Early results with the use of electrical stimulation are promising; however, prospective studies are needed to better define the role of this modality in managing these injuries. PMID- 11029562 TI - Opioid detoxification under anaesthesia. PMID- 11029563 TI - Sevoflurane progressively prolongs the QT interval in unpremedicated female adults. AB - Thirty-six unpremedicated women due for gynaecological surgery were examined for time-dependent prolongation of the QT interval in the electrocardiogram (ECG) before and after induction of anaesthesia using either sevoflurane or propofol. The conventional inhalational technique to induce anaesthesia with sevoflurane. ECG recordings were taken before, 2, 5 and 10 min after drug administration. Sevoflurane significantly lengthened (P < 0.001) the heart rate corrected QT interval within 10 min from 434 +/- 5 ms to 459 +/- 6 ms (mean +/- SEM). Already after 2 min of sevoflurane application a trend towards prolongation was visible. The critical value of 440 ms in the rate-corrected QT interval was exceeded in four patients in the sevoflurane group (n = 18) but in only in one patient in the propofol group (n = 18). Rate-corrected QT interval prolongation caused by sevoflurane needs to be recognized early in order to prevent the critical ventricular tachycardia torsade de pointes. PMID- 11029564 TI - Elimination of procalcitonin and plasma concentrations during continuous veno venous haemodiafiltration in septic patients. AB - The elimination of procalcitonin and the course of plasma concentrations during continuous veno-venous haemodiafiltration were measured in patients with sepsis or multiple organ dysfunction syndrome, because these patients are a main target group for the measurement of procalcitonin and often require renal replacement therapy. Procalcitonin was measured in the prefilter plasma and the filtrate at 5 min, 15 min and 1, 2, 4, 6, 12, 24 h after set-up of continuous veno-venous haemodiafiltration. In a prospective study, 19 patients with plasma levels of procalcitonin > 3 ng mL-1 and acute oliguric renal failure treated with continuous veno-venous haemodiafiltration using a polysulphone membrane, were evaluated for the study of clearance. Twenty-one control patients (procalcitonin < 2 ng mL-1) were studied to determine whether filtration itself induced a procalcitonin response. No interventions were required. In patients with low procalcitonin concentrations (procalcitonin < 2 ng mL-1) continuous veno-venous haemodiafiltration did not cause a rise in procalcitonin. In patients with increased procalcitonin plasma concentrations (> 3 ng mL-1), the protein was removed through the polysulphone membrane, with a final clearance of 4 mL min-1 after the initial adsorption period (clearance 0.4-0.9 mL min-1 during the first hour of continuous veno-venous haemodiafiltration). Thus, on the average, approximately 10% of plasma concentrations were measurable in the filtrate ultimately. However, procalcitonin plasma levels were not significantly altered during continuous veno-venous haemodiafiltration (86% of the initial concentration after 24 h). Although procalcitonin is removed from the plasma during continuous veno-venous haemodiafiltration in measurable amounts plasma procalcitonin concentrations did not change significantly during haemodiafiltration. Procalcitonin thus can also be used as a diagnostic parameter in patients undergoing continuous veno-venous haemodiafiltration. PMID- 11029565 TI - Lidocaine inhalation for local anaesthesia and attenuation of bronchial hyper reactivity with least airway irritation. Effect of three different dose regimens. AB - The inhalation of lidocaine attenuates bronchial hyper-reactivity but also causes airway irritation. However, how lidocaine dose and plasma concentration influence relationships are unknown. Accordingly, we evaluated the effects of three concentrations of lidocaine (1, 4, and 10%, total dose of 0.5, 2.0, and 5.0 mg kg 1, respectively) vs. placebo in 15 mild asthmatic patients, selected by their response to a histamine challenge (decrease in FEV1 > 20% to less than 18 mg mL-1 of histamine [PC20]). Baseline lung function, histamine-induced bronchoconstriction, topical anaesthesia, and lidocaine plasma concentrations were obtained. FEV1 following lidocaine inhalation showed the greatest decrease for the highest dose (from 3.79 +/- 0.15-3.60 +/- 0.15; P = 0.0012). Lidocaine inhalation increased baseline PC20 (6.1 +/- 1.3 mg mL-1) significantly (to 11.8 +/- 3.1, 16.1 +/- 3.3, and 18.3 +/- 4.5 mg mL-1, respectively) with no difference between the two highest doses. The duration of local anaesthesia was not significantly different between lidocaine concentrations of 4% and 10%. Thus, lidocaine inhalation, with increasing concentrations of the aerosolized solution, increases initial bronchoconstriction while significant attenuation of bronchial hyper-reactivity is not further enhanced with increasing concentrations from 4 to 10%. Plasma concentrations of lidocaine were always far below the toxic threshold. In conclusion, when local anaesthesia of the airways is required a lidocaine dose of 2.0 mg kg-1 as a 4% solution can be recommended for local anaesthesia and attenuation of bronchial hyper-reactivity with the least airway irritation. PMID- 11029566 TI - A central venous catheter coated with benzalkonium chloride for the prevention of catheter-related microbial colonization. AB - In an attempt to overcome infections associated with central venous catheters, a new antiseptic central venous catheter coated with benzalkonium chloride on the internal and external surfaces has been developed and evaluated in a clinical trial. Patients (235) randomly received either a triple-lumen central venous catheter coated with benzalkonium chloride (117) or a polyurethane non-antiseptic catheter (118). The incidence of microbial colonization of both catheters and retained antiseptic activity of the benzalkonium chloride device following removal were determined. The benzalkonium chloride resulted in a significant reduction of the incidence of microbial colonization on both the internal and external catheter surfaces. The reduction in colonization was detected at both the intradermal (21 benzalkonium chloride catheters vs. 38 controls, P = 0.0016) and distal segments of the antiseptic-coated catheters. Following catheter removal retained activity was demonstrated in benzalkonium chloride catheters which had been in place for up to 12 days. No patients developed adverse reactions to the benzalkonium chloride catheters. The findings demonstrate that the benzalkonium chloride catheter significantly reduced the incidence of catheter-associated colonization. PMID- 11029567 TI - Bedside percutaneous tracheostomy experience with 72 critically ill patients. AB - Tracheostomy is necessary in intensive care unit (ICU) patients requiring prolonged mechanical ventilation. As an alternative to the standard surgical method, percutaneous techniques are available. Seventy-two patients were electively selected for percutaneous tracheostomy (PCT) in a nine-bed combined medical-surgical intensive care unit. PCT was performed at bedside with the Portex Percutaneous Tracheostomy Kit that uses the Griggs technique. The procedure time and early complications were recorded. The procedure was successful in all patients. The average duration of placement was 7.4 min. There were no tracheostomy-related deaths. Major bleeding occurred in three patients and required surgical intervention. In one patient, minor bleeding occurred at the stoma site that resolved with applied pressure. Wound infections were treated with local antiseptics in two patients. These findings suggest that PCT is a simple, quick and safe procedure. PMID- 11029568 TI - Epidural fentanyl-bupivacaine compared with clonidine-bupivacaine for analgesia in labour. AB - Alpha-adrenergic agonists produce pain relief through an opioid independent mechanism and may be alternatives to opioids for combination with local anaesthetics for analgesia during labour. We studied 41 pregnant women. Epidural block was performed with 75 microg clonidine (n = 20) or 50 microg fentanyl (n = 21) combined with 0.125% bupivacaine (10 mL). Maternal vital parameters were measured. Analgesia was evaluated using a visual analogue scale (VAS); sedation was scored using a five-point scale. There were no differences in maternal vital parameters, fetal heart rate (FHR) or Apgar scores between the groups. Analgesia lasted longer in the bupivacaine-clonidine group (139.4 +/- 31 min) compared with the bupivacaine-fentanyl group (127.9 +/- 48 min) (P = 0.42). Additional analgesic requirement was more often in the fentanyl-bupivacaine group and total bupivacaine requirement was less in the clonidine-bupivacaine group (22.5 +/- 12.5 mg vs. 30.9 +/- 12.8 mg) (P = 0.04). This small study confirms that this combination of bupivacaine and clonidine provides satisfactory analgesia for first-stage labour, and of longer duration than bupivacaine-fentanyl. PMID- 11029569 TI - The accuracy of the estimation of body weight and height in the intensive care unit. AB - A telephone survey of 20 English intensive care units (ICUs) confirmed that visual estimation of patient weight is often performed. Four experienced intensive care staff (three doctors and one nurse) estimated the weight and measured the height of 30 volunteers and the estimates were compared with accurate reference measurements. The estimates were shown to be significantly inaccurate for individual observers. We consider the degree of inaccuracy to be of clinical importance. However, pooling the individual estimates of weight as mean values improved accuracy. Recommendations are made concerning the performance of height and weight determination. PMID- 11029571 TI - Near-fatal acute dissection of the thoracic aorta involving the right coronary artery. AB - We describe a patient with aortic dissection extending into the right coronary artery who was successfully resuscitated and operated upon after a cardiac arrest during transfer to the operating room. The use of transoesophageal echocardiography was particularly helpful for rapid diagnosis of aortic type A dissection and urgent surgical treatment. PMID- 11029570 TI - Pulmonary lipiodol embolism during transcatheter arterial chemoembolization for hepatoblastoma under general anaesthesia. AB - We present a case of pulmonary embolism that occurred during the injection of lipiodol during transcatheter arterial chemoembolization under general anaesthesia. A 7-year-old child suffering from a large hepatoblastoma was admitted for arterial chemoembolization and carcinostatic administration. Pulmonary embolism due to lipiodol during arterial chemoembolization was evident by a sudden fall in oxyhaemoglobin saturation from 100 to 90%. This was associated with a spread of lipiodol into both lungs, particularly the middle lung zones and detected by chest fluoroscopy. Arterial blood gases returned to normal values 1 day later but pulmonary infiltration persisted for 7 days before final clearance. Pulmonary embolism caused by lipiodol during arterial chemoembolization is infrequent, but such a complication could prove fatal. Understanding the risk of pulmonary embolism in patients receiving lipiodol, during and after arterial chemoembolization, and late onset pulmonary injury is important and a close follow-up for several days after arterial chemoembolization is advisable. PMID- 11029572 TI - A successfully resuscitated case of amniotic fluid embolism. PMID- 11029573 TI - Synthesis of the translational apparatus is regulated at the translational level. AB - The synthesis of many mammalian proteins associated with the translational apparatus is selectively regulated by mitogenic and nutritional stimuli, at the translational level. The apparent advantages of the regulation of gene expression at the translational level are the speed and the readily reversible nature of the response to altering physiological conditions. These two features enable cells to rapidly repress the biosynthesis of the translational machinery upon shortage of amino acids or growth arrest, thus rapidly blocking unnecessary energy wastage. Likewise, when amino acids are replenished or mitogenic stimulation is applied, then cells can rapidly respond in resuming the costly biosynthesis of the translational apparatus. A structural hallmark, common to mRNAs encoding many components of the translational machinery, is the presence of a 5' terminal oligopyrimidine tract (5'TOP), referred to as TOP mRNAs. This structural motif comprises the core of the translational cis-regulatory element of these mRNAs. The present review focuses on the mechanism underlying the translational control of TOP mRNAs upon growth and nutritional stimuli. A special emphasis is put on the pivotal role played by ribosomal protein S6 kinase (S6K) in this mode of regulation, and the upstream regulatory pathways, which might be engaged in transducing external signals into activation of S6K. Finally, the possible involvement of pyrimidine-binding proteins in the translational control of TOP mRNAs is discussed. PMID- 11029574 TI - Characterization of acid-induced unfolding intermediates of glucose/xylose isomerase. AB - Acid-induced unfolding of the tetrameric glucose/xylose isomerase (GXI) from Streptomyces sp. NCIM 2730 has been investigated using intrinsic fluorescence, fluorescence quenching, second derivative spectroscopy, hydrophobic dye (1 anilino-8-naphthalene-sulfonate) binding and CD techniques. The pH dependence of tryptophanyl fluorescence of GXI at different temperatures indicated the presence of two stable intermediates at pH 5.0 and pH 3.0. The pH 3.2 intermediate was a dimer and exhibited molten globule-like characteristics, such as the presence of native-like secondary structure, loss of tertiary structure, increased exposure of hydrophobic pockets, altered microenvironment of tyrosine residues and increased accessibility to quenching by acrylamide. Fluorescence and CD studies on GXI at pH 5.0 suggested the involvement of a partially folded intermediate state in the native to molten globule state transition. The partially folded intermediate state retained considerable secondary and tertiary structure compared to the molten globule state. This state was characterized by its hydrophobic dye binding capacity, which is smaller than the molten globule state, but was greater than that of the native state. This state shared the dimeric status of the molten globule state but was prone to aggregate formation as evident by the Rayleigh light scattering studies. Based on these results, the unfolding pathway of GXI can be illustrated as: N-->PFI-->MG-->U; where N is the native state at pH 7.5; PFI is the partially folded intermediate state at pH 5.0; MG is the molten globule state at pH 3.2 and U is the monomeric unfolded state of GXI obtained in the presence of 6 M GdnHCl. Our results demonstrate the existence of a partially folded state and molten globule state on the unfolding pathway of a multimeric alpha/beta barrel protein. PMID- 11029575 TI - Solubilization, partial purification and photolabeling of the integral membrane protein lysophospholipid:acyl-CoA acyltransferase (LAT). AB - In the present study, we defined experimental conditions that allowed the extraction of the integral membrane protein lysophospholipid:acyl-CoA acyltransferase (LAT, EC 2.3.1.23) from membranes while maintaining the full enzyme activity using the nonionic detergent n-octyl glucopyranoside (OGP) and solutions of high ionic strength. We found that the optimal OGP concentration depended on the ionic strength of the solubilization buffer. Fluorescence measurements with 1,6-diphenyl-1,3,5-hexatriene indicated that the critical micellar concentration (CMC) of OGP decreased with increasing salt concentrations. Analogous studies revealed that the zwitterionic detergent Chaps was ineffective in extracting LAT from membranes in the absence of salt, whereas its solubilization efficiency increased with increasing salt concentrations. Detailed lipid analysis of the different protein/lipid/detergent mixed micelles showed that the protein/lipid/OGP mixed micelles were relatively enriched with sphingomyelin (SPM) compared to protein/lipid/Chaps mixed micelles, indicating that the differences in the solubilization efficiency may be due to the ability to extract more SPM from membranes. When the protein/lipid/OGP mixed micelles were dissociated into protein/detergent and lipid/detergent complexes by the addition of increasing Chaps concentrations, one-tenth of the LAT enzyme activity was preserved making the enzyme accessible to protein purification. Analysis by native PAGE revealed that in the presence of excess Chaps a high molecular mass protein complex migrated into the gel which could be photolabeled by 125I labelled-18-(4'-azido-2'-hydroxybenzoylamino)-oleyl-CoA. This fatty acid analogue has been shown to be a competitive inhibitor of LAT enzyme activity in the dark, and an irreversible inhibitor after photolysis. Therefore, this protein complex is assumed to contain the LAT enzyme. PMID- 11029576 TI - Identification of neoxanthin synthase as a carotenoid cyclase paralog. AB - Neoxanthin, a precursor of the plant hormone abscisic acid, is an allenic xanthophyll recognized as the last product of carotenoid synthesis in green plants. A cDNA for neoxanthin synthase (NSY) was isolated from tomato using a molecular approach based on the mechanistic and structural similarities of NSY to two other closely related carotenogenic enzymes, lycopene cyclase (LCY) and capsanthin-capsorubin synthase (CCS). The identified tomato NSY cDNA (T.NSY) encodes a 56-kDa plastid-targeted protein that when expressed in Escherichia coli, catalyzes the conversion of violaxanthin to neoxanthin. In tobacco leaves that transiently express T.NSY, an increase in neoxanthin content with a concomitant decrease in violaxanthin is observed. NSY is structurally similar to LCY and CCS. However, in Cyanobacteria, the generally accepted progenitor of plastids, both CCS and NSY are absent while LCY is present. LCY catalyzes a simplified version of the reaction catalyzed by NSY and CCS suggesting that these two enzymes were remodeled from LCY during higher plant evolution to create new forms of oxidized carotenoids. PMID- 11029577 TI - Effect of amino-acid substitutions on Alzheimer's amyloid-beta peptide glycosaminoglycan interactions. AB - One of the major clinical features of Alzheimer's disease is the presence of extracellular amyloid plaques that are associated with glycosaminoglycan containing proteoglycans. It has been proposed that proteoglycans and glycosaminoglycans facilitate amyloid fibril formation and/or stabilize these aggregates. Characterization of proteoglycan-protein interactions has suggested that basic amino acids in a specific conformation are necessary for glycosaminoglycan binding. Amyloid-beta peptide (Abeta) has a cluster of basic amino acids at the N-terminus (residues 13-16, His-His-Gln-Lys), which are considered critical for glycosaminoglycan interactions. To understand the molecular recognition of glycosaminoglycans by Abeta, we have examined a series of synthetic peptides with systematic alanine substitutions. These include: His13 ->Ala, His14-->Ala, Lys16-->Ala, His13His14Lys16-->Ala and Arg5His6-->Ala. Alanine substitutions result in differences in both the secondary and fibrous structure of Abeta1-28 as determined by circular dichroism spectroscopy and electron microscopy. The results demonstrate that the His-His-Gln-Lys region of Abeta, and in particular His13, is an important structural domain, as Ala substitution produces a dysfunctional folding mutant. Interaction of the substituted peptides with heparin and chondroitin sulfate glycosaminoglycans demonstrate that although electrostatic interactions contribute to binding, nonionic interactions such as hydrogen bonding and van der Waals packing play a role in glycosaminoglycan-induced Abeta folding and aggregation. PMID- 11029578 TI - Preferential interaction of alpha-tocopherol with phosphatidylcholines in mixed aqueous dispersions of phosphatidylcholine and phosphatidylethanolamine. AB - The effect of alpha-tocopherol on the structure and phase behaviour of mixed aqueous dispersions of phosphatidylcholine and phosphatidylethanolamine has been examined by synchrotron X-ray diffraction and freeze-fracture electron microscopy. Equimolar mixtures of fully saturated derivatives of phospholipids that show gel phase immiscibility were examined including dimyristoylglycerophosphocholine/dipalmitoylglycerophosphoethanolamin e and distearoylglycerophosphocholine/dilauroylglycerophosphoethanolamine++ +. Analysis of the X-ray scattering intensities recorded at wide angles during heating scans of mixed aqueous dispersions containing 2.5 or 5 mol% alpha-tocopherol showed that alpha-tocopherol disordered the acyl chain packing of the phosphatidylcholine to a greater extent than the phosphatidylethanolamine component of the mixture. This suggested that alpha-tocopherol preferentially interacts with phosphatidylcholine rather than phosphatidylethanolamine, irrespective of whether this was the high or low melting point component of the mixture. The presence of 20 mol% alpha-tocopherol in either phospholipid mixture prevented gel phase separation during the prior cooling scan and no conclusions could be drawn as to the distribution of alpha-tocopherol in these mixtures. PMID- 11029579 TI - Biochemical analysis of a thermostable tryptophan synthase from a hyperthermophilic archaeon. AB - Pyridoxal 5'-phosphate-dependent tryptophan synthase catalyzes the last two reactions of tryptophan biosynthesis, and is comprised of two distinct subunits, alpha and beta. TktrpA and TktrpB, which encode the alpha subunit and beta subunit of tryptophan synthase from a hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1, were independently expressed in Escherichia coli and their protein products were purified. Tryptophan synthase complex (Tk-TS complex), obtained by heat treatment of a mixture of the cell-free extracts containing each subunit, was also purified. Gel-filtration chromatography revealed that Tk-TrpA was a monomer (alpha), Tk-TrpB was a dimer (beta2), and Tk-TS complex was a tetramer (alpha2 beta2). The Tk-TS complex catalyzed the overall alphabeta reaction with a specific activity of 110 micromol Trp per micromol active site per min under its optimal conditions (80 degrees C, pH 8.5). Individual activity of the alpha and beta reactions of the Tk-TS complex were 8.5 micromol indole per micromol active site per min (70 degrees C, pH 7.0) and 119 micromol Trp per micromol active site per min (90 degrees C, pH 7.0), respectively. The low activity of the alpha reaction of the Tk-TS complex indicated that turnover of the beta reaction, namely the consumption of indole, was necessary for efficient progression of the alpha reaction. The alpha and beta reaction activities of independently purified Tk-TrpA and Tk-TrpB were 10-fold lower than the respective activities detected from the Tk-TS complex, indicating that during heat treatment, each subunit was necessary for the other to obtain a proper conformation for high enzyme activity. Tk-TrpA showed only trace activities at all temperatures examined (40-85 degrees C). Tk-TrpB also displayed low levels of activity at temperatures below 70 degrees C. However, Tk-TrpB activity increased at temperatures above 70 degrees C, and eventually at 100 degrees C, reached an equivalent level of activity with the beta reaction activity of Tk-TS complex. Taking into account the results of circular dichroism analyses of the three enzymes, a model is proposed which explains the relationship between structure and activity of the alpha and beta subunits with changes in temperature. This is the first report of an archaeal tryptophan synthase, and the first biochemical analysis of a thermostable tryptophan synthase at high temperature. PMID- 11029580 TI - Prostromelysin-1 (proMMP-3) stimulates plasminogen activation by tissue-type plasminogen activator. AB - Matrix metalloproteinase-3 (MMP-3 or stromelysin-1) specifically binds to tissue type plasminogen activator (t-PA), without however, hydrolyzing the protein. Binding affinity to proMMP-3 is similar to single chain t-PA, two chain t-PA and active site mutagenized t-PA (Ka of 6.3 x 106 to 8.0 x 106 M-1), but is reduced for t-PA lacking the finger and growth factor domains (Ka of 2.0 x 106 M-1). Activation of native Glu-plasminogen by t-PA in the presence of proMMP-3 obeys Michaelis-Menten kinetics; at saturating concentrations of proMMP-3, the catalytic efficiency of two chain t-PA is enhanced 20-fold (kcat/Km of 7.9 x 10-3 vs. 4.1 x 10-4 microM-1.s-1). This is mainly the result of an enhanced affinity of t-PA for its substrate (Km of 1.6 microM vs. 89 microM in the absence of proMMP-3), whereas the kcat is less affected (kcat of 1.3 x 10-2 vs. 3.6 x 10-2 s 1). Activation of Lys-plasminogen by two chain t-PA is stimulated about 13-fold at a saturating concentration of proMMP-3, whereas that of miniplasminogen is virtually unaffected (1.4-fold). Plasminogen activation by single chain t-PA is stimulated about ninefold by proMMP-3, whereas that by the mutant lacking finger and growth factor domains is stimulated only threefold. Biospecific interaction analysis revealed binding of Lys-plasminogen to proMMP-3 with 18-fold higher affinity (Ka of 22 x 106 M-1) and of miniplasminogen with fivefold lower affinity (Ka of 0.26 x 106 M-1) as compared to Glu-plasminogen (Ka of 1.2 x 106 M-1). Plasminogen and t-PA appear to bind to different sites on proMMP-3. These data are compatible with a model in which both plasminogen and t-PA bind to proMMP-3, resulting in a cyclic ternary complex in which t-PA has an enhanced affinity for plasminogen, which may be in a Lys-plasminogen-like conformation. Maximal binding and stimulation require the N-terminal finger and growth factor domains of t-PA and the N-terminal kringle domains of plasminogen. PMID- 11029581 TI - Sterol-induced upregulation of phosphatidylcholine synthesis in cultured fibroblasts is affected by the double-bond position in the sterol tetracyclic ring structure. AB - We have examined how a specific enrichment of cultured fibroblasts with various sterols (cholesterol, lathosterol, 7-dehydrocholesterol, allocholesterol and dihydrocholesterol) regulate synthesis de novo of phosphatidylcholine, cholesterol and cholesteryl (or steryl) esters in human skin fibroblasts. When human skin fibroblasts were incubated for 1 h with 130 microM cholesterol/CyD complexes, the mass of cellular free cholesterol increased by 100 nmol.mg-1 protein (from 90 nmol.mg-1 to 190 nmol.mg-1 protein). A similar exposure of cells to different sterol/CyD complexes increased the cell sterol content between 38 and 181 nmol sterol per mg cell protein. In cholesterol-enriched cells, the rate of phosphatidylcholine synthesis was doubled compared to control cells, irrespective of the type of precursor used ([3H]choline, [3H]palmitic acid, or [14C]glycerol). Enrichment of fibroblasts with 7-dehydrocholesterol, allocholesterol, or dihydrocholesterol also upregulated phosphatidylcholine synthesis, whereas cells enriched with lathosterol failed to upregulate their phosphatidylcholine synthesis. The activity of membrane-bound CTP:phosphocholine cytidylyltransferase, the rate-limiting enzyme, was increased by 47 +/- 4% in cholesterol-enriched cells whereas its activity was unchanged in lathosterol enriched cells. Sterol enrichment with all tested sterols (including lathosterol) down-regulated acetate-incorporation into cholesterol, and upregulated sterol esterification in the sterol-enriched fibroblasts. Using 31P-NMR to measure the lamellar-to-hexagonal (Lalpha-HII) phase transition in multilamellar lipid dispersions, lathosterol-containing membranes underwent their transition at significantly higher temperatures compared to membranes containing any of the other sterols. In a system with 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphoethanolamine and either cholesterol or lathosterol (70:30 mol/mol), differential scanning calorimetry also revealed that the Lalpha-HII-transition occurred at a higher temperature with lathosterol compared to either cholesterol, allocholesterol, or dihydrocholesterol. These findings together suggest that there may exist a correlation between the propensity of a sterol to stabilize the Lalpha-HII-transition and its capacity to upregulate the activity of CTP:phosphocholine cytidylyltransferase in cells. PMID- 11029582 TI - Structural domains involved in the RNA folding activity of RNA helicase II/Gu protein. AB - RNA helicase II/Gu (RH II/Gu) is a nucleolar protein that unwinds dsRNA in a 5' to 3' direction, and introduces a secondary structure into a ssRNA. The helicase domain is at the N-terminal three-quarters of the molecule and the foldase domain is at the C-terminal quarter. The RNA folding activity of RH II/Gu is not a mere artifact of its binding to RNA. This study narrows down the RNA foldase domain to amino acids 749-801 at the C-terminus of the protein. Dissection of this region by deletion and site-directed mutagenesis shows that the four FRGQR repeats, as well as the C-terminal end bind RNA independently. These juxtaposed subdomains are both important for the RNA foldase activity of RH II/Gu. Mutation of either repeat 2 or repeat 4, or simultaneous mutation of Lys792, Arg793 and Lys797 at the C-terminal end of RH II/Gu to alanines inhibits RNA foldase activity. The last 17 amino acids of RH II/Gu can be replaced by an RNA binding motif from nucleolar protein p120 without a deleterious effect on its foldase activity. A model is proposed to explain how RH II/Gu binds and folds an RNA substrate. PMID- 11029583 TI - Dissociation and unfolding of cold-active alkaline phosphatase from atlantic cod in the presence of guanidinium chloride. AB - Cold-adaptation of enzymes involves improvements in catalytic efficiency. This paper describes studies on the conformational stability of a cold-active alkaline phosphatase (AP) from Atlantic cod, with the aim of understanding more clearly its structural stability in terms of subunit dissociation and unfolding of monomers. AP is a homodimeric enzyme that is only active in the dimeric state. Tryptophan fluorescence, size-exclusion chromatography and enzyme activity were used to monitor alterations in conformational state induced by guanidinium chloride or urea. In cod AP, a clear distinction could be made between dissociation of dimers into monomers and subsequent unfolding of monomers (fits a three-state model). In contrast, dimer dissociation of calf AP coincided with the monophasic unfolding curve observed by tryptophan fluorescence (fits a two-state model). The DeltaG for dimer dissociation of cod AP was 8.3 kcal.mol-1, and the monomer stabilization free energy was 2.2 kcal.mol-1, giving a total of 12.7 kcal.mol-1, whereas the total free energy of calf intestinal AP was 17.3 kcal.mol 1. Thus, dimer formation provided a major contribution to the overall stability of the cod enzyme. Phosphate, the reaction product, had the effect of promoting dimer dissociation and stabilizing the monomers. Cod AP has reduced affinity for inorganic phosphate, the release of which is the rate-limiting step of the reaction mechanism. More flexible links at the interface between the dimer subunits may ease structural rearrangements that facilitate more rapid release of phosphate, and thus catalytic turnover. PMID- 11029584 TI - Domains of Brn-2 that mediate homodimerization and interaction with general and melanocytic transcription factors. AB - The class III POU gene brn-2, encoding the Brn-2/N-Oct-3 transcription factor, is widely expressed in the developing mammalian central nervous system. Brn-2 has also been found to regulate the melanocytic phenotype with N-Oct-3 DNA binding activity elevated in malignant melanoma, however, its mode of action is yet to be defined. The functional role of the Brn-2 transcription factor has been investigated through the analysis of protein-protein interactions it forms with a number of basal and melanocytic transcriptional regulatory proteins. In vivo interactions were tested by gene-cotransfection using the mammalian GAL4-Herpes Simplex viral protein 16 (VP16) two hybrid formation and direct protein binding by in vitro glutathione S-transferase (GST)-pull down assay. The Brn-2 protein was found to homodimerize in vivo with high affinity, using Brn-2 deletion constructs dimer complex formation was found to be dependent on the presence of both the homeodomain and linker regions of the POU-domain. However, the POU homoedomain was dispensable for the formation of the dimerization interface in one of the partner molecules but not both, when the POU-linker region was removed the ability to interact was lost irrespective of the presence of the homeodomain. Dimerization of Brn-2/N-Oct-3 was also found to occur in DNA binding assays using melanoma cell line nuclear extracts and a recently reported dimer target sequence probe, which may have significant consequences for gene regulation in melanocytic tumours. Low affinity Brn-2 protein contacts have also been found with the basal transcription complex, including TATA binding protein (TBP) and the transcriptional coactivator p300, and with the Sox-10 and Pax-3 transcription factors that are known to play an important role in melanocyte cell formation. PMID- 11029585 TI - A novel mammalian Smt3-specific isopeptidase 1 (SMT3IP1) localized in the nucleolus at interphase. AB - A novel Smt3-specific isopeptidase, SMT3IP1, was cloned using a yeast two-hybrid screen with Smt3b as bait. The clone, named SMT3IP1 (Smt3-specific isopeptidase 1), which bound to Smt3b but not SUMO-1 in the two-hybrid system, was distantly related to budding yeast Saccharomyces cerevisiae Ulp1, human SENP1 or human SUSP1. The catalytic domains in the C-terminal region were very similar, but the N-terminal region was quite different to other enzymes. The cysteine, histidine and asparatic acid residues in the catalytic domains were conserved. SMT3IP1 expressed by the baculovirus-expression system had the ability to cleave SUMO-1 or Smt3b from SUMO-1/RanGAP1 or Smt3b/RanGAP1 conjugates, respectively, and the activity was a little stronger towards the Smt3b conjugate than towards the SUMO 1 conjugate. Furthermore, the enzyme bound more strongly to Smt3a and Smt3b than to SUMO-1 in vitro. The enzyme did not cleave Nedd8 from Nedd8/cullin-1. Nor did it cleave ubiquitin from ubiquitinated p53. SMT3IP1 was localized almost exclusively at the nucleolus during interphase. The N-terminal sequence was responsible for the nucleolar localization of this enzyme. Whether SMT3IP1 functions in the nucleolus or just stays there before it functions in the nucleus, as shown in the case of CDC14 phosphatase, remains to be elucidated. PMID- 11029586 TI - Coordinate up- and down-regulation of glutathione-dependent prostaglandin E synthase and cyclooxygenase-2 in A549 cells. Inhibition by NS-398 and leukotriene C4. AB - Recently, a microsomal protein with 38% sequence identity to microsomal glutathione S-transferase 1 was shown to constitute an inducible, glutathione dependent prostaglandin E synthase (PGES). To investigate the relationship between cyclooxygenase and PGES, a time-course study on protein expression was performed in A549 cells after treatment with interleukin-1beta. The result demonstrated a tandem expression of cyclooxygenase-2 and PGES. The observed induction of PGES protein correlated with microsomal PGES activity. No comparable PGES activity was observed in the absence of glutathione or in the cytosolic fraction. In addition, tumour necrosis factor-alpha was found to induce PGES in these cells. Dexamethasone was found to completely suppress the effect of both cytokines on PGES induction. We also describe a quantitative method, based on RP HPLC with UV detection for the measurements of PGES activity. This method was used to screen potential PGES inhibitors. Several nonsteroidal anti-inflammatory drugs, stable prostaglandin H2 analogues and cysteinyl leukotrienes were screened for inhibition of PGES activity. NS-398, sulindac sulfide and leukotriene C4 were all found to inhibit PGES activity with IC50 values of 20 microM, 80 microM and 5 microM, respectively. In conclusion, it appears that PGES and cyclooxygenase-2 are functionally coupled in A549 cells and that a required coordinate expression of these enzymes allows for efficient biosynthesis of prostaglandin E2. PMID- 11029587 TI - Butyrate metabolism upstream and downstream acetyl-CoA synthesis and growth control of human colon carcinoma cells. AB - Butyrate is a short chain fatty acid (SCFA) produced by bacterial fermentation of dietary fibers in the colon lumen which severely affects the proliferation of colon cancer cells in in vitro experiments. Although butyrate is able to interfere with numerous cellular targets including cell cycle regulator expression, little is known about butyrate metabolism and its possible involvement in its effect upon colon carcinoma cell growth. In this study, we found that HT-29 Glc-/+ cells strongly accumulated and oxidized sodium butyrate without producing ketone bodies, nor modifying oxygen consumption nor mitochondrial ATP synthesis. HT-29 cells accumulated and oxidized sodium acetate at a higher level than butyrate. However, sodium butyrate, but not sodium acetate, reduced cell growth and increased the expression of the cell cycle effector cyclin D3 and the inhibitor of the G1/S cdk-cyclin complexes p21/WAF1/Cip1, demonstrating that butyrate metabolism downstream of acetyl-CoA synthesis is not required for the growth-restraining effect of this SCFA. Furthermore, HT-29 cells modestly incorporated the 14C-labelled carbon from sodium butyrate into cellular triacylglycerols and phospholipids. This incorporation was greatly increased when D-glucose was present in the incubation medium, corresponding to the capacity of hexose to circulate in the pentose phosphate pathway allowing NADPH synthesis required for lipogenesis. Interestingly, when HT-29 cells were cultured in the presence of sodium butyrate, their capacity to incorporate 14C-labelled sodium butyrate into triacylglycerols and phospholipids was increased more than twofold. In such experimental conditions, HT-29 cells when observed under an electronic microscope, were found to be characterized by an accumulation of lipid droplets in the cytosol. Our data strongly suggest that butyrate acts upon colon carcinoma cells upstream of acetyl CoA synthesis. In contrast, the metabolism downstream of acetyl-CoA [i.e. oxidation in the tricarboxylic acid (TCA) cycle and lipid synthesis] likely acts as a regulator of butyrate intracellular concentration. PMID- 11029588 TI - Topology and proximity relationships of yeast mitochondrial ATP synthase subunit 8 determined by unique introduced cysteine residues. AB - We have used site-directed chemical labelling to demonstrate the membrane topology and to identify neighbouring subunits of subunit 8 (Y8) in yeast mitochondrial ATP synthase (mtATPase). Unique cysteine residues were introduced at the N or C-terminus of Y8 by site-directed mutagenesis. Expression and targeting to mitochondria in vivo of each of these variants in a yeast Y8 null mutant was able to restore activity to an otherwise nonfunctional ATP synthase complex. The position of each introduced cysteine relative to the inner mitochondrial membrane was probed with thiol-specific nonpermeant and permeant reagents in both intact and lysed mitochondria. The data indicate that the N terminus of Y8 is located in the intermembrane space of mitochondria whereas the C-terminus is located within the mitochondrial matrix. The proximity of Y8 to other proteins of mtATPase was tested using heterobifunctional cross-linking reagents, each with one thiol-specific reactive group and one nonspecific, photoactivatible reactive group. These experiments revealed the proximity of the C-terminal domain of Y8 to subunits d and f, and that of the N-terminal domain to subunit f. It is concluded that Y8 possesses a single transmembrane domain which extends across the inner membrane of intact mitochondria. As subunit d is a likely component of the stator stalk of mitochondrial ATP synthase, we propose, on the basis of the observed cross-links, that Y8 may also be part of the stator stalk. PMID- 11029589 TI - Developmental and environmental regulation of antifreeze proteins in the mealworm beetle Tenebrio molitor. AB - The yellow mealworm beetle, Tenebrio molitor, contains a family of small Cys-rich and Thr-rich thermal hysteresis proteins that depress the hemolymph freezing point below the melting point by as much as 5. 5 degrees C (DeltaT = thermal hysteresis). Thermal hysteresis protein expression was evaluated throughout development and after exposure to altered environmental conditions. Under favorable growth conditions, small larvae (11-13 mg) had only low levels of thermal hysteresis proteins or thermal hysteresis protein message, but these levels increased 10-fold and 18-fold, respectively, by the final larval instar (> 190 mg), resulting in thermal hysteresis > 3 degrees C. Exposure of small larvae (11-13 mg) to 4 weeks of cold (4 degrees C) caused an approximately 20-fold increase in thermal hysteresis protein concentration, well in excess of the less than threefold developmental increase seen after 4 weeks at 22 degrees C. Exposure of large larvae (100-120 mg) to cold caused 12-fold and sixfold increases in thermal hysteresis protein message and protein levels, respectively, approximately double the maximum levels they would have attained in the final larval instar at 22 degrees C. Thus, thermal hysteresis increased to similar levels (> 4 degrees C) in the cold, irrespective of the size of the larvae (the overwintering stage). At pupation, thermal hysteresis protein message levels decreased > 20-fold and remained low thereafter, but thermal hysteresis activity decreased much more slowly. Exposure to cold did not reverse this decline. Desiccation or starvation of larvae had comparable effects to cold exposure, but surprisingly, short daylength photoperiod or total darkness had no effect on either thermal hysteresis or message levels. As all environmental conditions that caused increased thermal hysteresis also inhibited growth, we postulate that developmental arrest is a primary factor in the regulation of T. molitor thermal hysteresis proteins. PMID- 11029590 TI - A novel extracellular esterase from Bacillus subtilis and its conversion to a monoacylglycerol hydrolase. AB - A novel gene lipB, which encodes an extracellular lipolytic enzyme, was identified in the Bacillus subtilis genomic DNA sequence. We have cloned and overexpressed lipB in B. subtilis and Escherichia coli and have also purified the enzyme from a B. subtilis culture supernatant to electrophoretic homogeneity. Four different lipase assays were used to determine its catalytic activity: pH stat, spectrophotometry, fluorimetry and the monomolecular film technique. LipB preferentially hydrolysed triacylglycerol-esters and p-nitrophenyl-esters of fatty acids with short chain lengths of 3Galbeta1-->4R is present on glycoprotein glycans. To investigate if alpha3-galactosylated glycoproteins, just like desialylated glycoproteins, could be cleared by the hepatic lectin, the affinities of alpha3-galactosylated compounds towards this lectin were determined using an in vitro inhibition assay, and were compared with those of the parent compounds terminating in Galbeta1- >4R. Diantennary, triantennary and tetraantennary oligosaccharides that form part of N-glycans were alpha3-galactosylated to completion by use of recombinant bovine alpha3-galactosyltransferase. Similarly, desialylated alpha1-acid glycoprotein (orosomucoid) was alpha3-galactosylated in vitro. The alpha3 galactosylation of a branched, Galbeta1-->4-terminated oligosaccharide lowered its affinity for the membrane-bound lectin on whole rat hepatocytes 50-250-fold, and for the detergent-solubilized hepatic lectin 7-50-fold. In contrast, alpha3 galactosylation of asialo-alpha1-acid glycoprotein caused only a minor decrease in affinity, increasing the IC50 from 5 to 15 nM. Fully alpha3-galactosylated alpha1-acid glycoprotein, intravenously injected into the mouse, was rapidly cleared from the circulation, with a clearance rate close to that of asialo alpha1-acid glycoprotein (t1/2 of 0.42 min vs. 0.95 min). Its uptake was efficiently inhibited by pre-injection of an excess asialo-fetuin. Organ distribution analysis showed that the injected alpha1-acid glycoprotein accumulated predominantly in the liver. Taken together, these observations suggest that serum glycoproteins that are heavily alpha3-galactosylated will be rapidly cleared from the bloodstream via the hepatic lectin. It is suggested that glycosyltransferase expression in murine hepatocytes is tightly regulated in order to prevent undesired uptake of hepatocyte-derived, circulating glycoproteins. PMID- 11029596 TI - Antioxidant therapy for AIDS. PMID- 11029597 TI - Effect of gemfibrozil on peripheral atherosclerosis and platelet activation in a pig model of hyperlipidemia. AB - BACKGROUND: Gemfibrozil has been shown to have beneficial effects in the primary and secondary prevention of atherosclerosis. However, a platelet pro-activating effect induced by the drug has been reported. MATERIAL AND METHODS: We analysed the effect of hyperlipidemia and its treatment with gemfibrozil on platelet fibrinogen binding and the development of early fibrinogen-rich vascular lesions in a porcine model of atherosclerosis. Polyclonal antibodies were raised against purified pig fibrinogen and intact platelets. Two groups of animals were fed a cholesterol/saturated fat-enriched diet for 50 days; one group was treated with gemfibrozil and the other with placebo. RESULTS: The hyperlipidemic diet induced a significant increase in total cholesterol; this was prevented by gemfibrozil (P<0.05). The increase in platelet-fibrinogen binding induced by hypercholesterolemia was mildly reduced in the gemfibrozil treated animals. Histological analysis of aortic vascular wall (abdominal aorta at the iliac bifurcation) from hyperlipidemic animals showed early lesions with fibrinogen infiltration; the lesions were reduced in the fibrate-treated animals. CONCLUSIONS: Gemfibrozil delayed the development of peripheral atherosclerotic plaque, normalised the impaired lipid profile induced by the hyperlipidemic diet and did not show a functionally detectable platelet pro-activating effect able to increase platelet-fibrinogen binding. PMID- 11029598 TI - ACE-inhibition modulates some endothelial functions in healthy subjects and in normotensive type 1 diabetic patients. AB - BACKGROUND: The usefulness of treatment with an angiotensin-converting enzyme inhibitor (ACE-inhibitor) in normotensive patients with type 1 diabetes is controversial. We investigated whether ACE-inhibition improves endothelial function in such patients and compared the responses to those in healthy subjects. DESIGN: We studied 23 healthy volunteers (controls, aged 29.8 [SD 7.0] years) and 24 type 1 diabetic patients (aged 28.7 [9. 6] years; HbA1c 8.1 [1.2]%; diabetes duration 13.8 [2-30] years; blood pressure < 140/90 mm Hg; 7 with microalbuminuria) after 5 weeks of ACE-inhibition (quinapril, 10 mg day-1) and placebo in a randomized, double-blind cross-over design. Estimates of endothelial function obtained were by flow-mediated vasodilation and plasma levels of endothelium-derived proteins. RESULTS: As estimated from the measurements on placebo, type 1 diabetic patients, as compared to the controls, had some impairment of endothelial function: plasma tissue-type plasminogen activator levels were lower (3.5 vs. 5.4 ng mL(-1), P<0.05), but there were no significant differences in brachial artery flow-mediated vasodilation or plasma levels of von Willebrand Factor, endothelin-1, plasminogen activator inhibitor-1, soluble E selectin or vascular cell adhesion molecule-1. As compared to placebo, ACE inhibition increased flow-mediated vasodilation in controls (by 3.84% points [95% CI, 0.66 - 7.02], P<0.05), but not in type 1 diabetic patients (0.82% points [95% CI, -2.72 - 4.36], P = 0.64; P = 0.08 vs. controls). On ACE-inhibition soluble E selectin levels decreased both in controls (from 43.0 to 37.0 ng mL(-1), P<0.01) and in type 1 diabetic patients (from 41.0 to 39.0 ng mL(-1), P = 0.09). Other endothelial markers did not change during ACE-inhibition. CONCLUSION: Normotensive type 1 diabetic patients with normoalbuminara or microalbuminuria have mild endothelial dysfunction. Short-term ACE-inhibition improves endothelial function as reflected by a decreased sE-selectin in healthy subjects and in normotensive type 1 diabetic patients. In healthy subjects, ACE-inhibition increases flow-mediated vasodilation. In contrast, in type 1 diabetic patients, ACE-inhibition does not affect flow-mediated vasodilation. PMID- 11029599 TI - Circulating concentrations of C-reactive protein and total sialic acid in tobacco smokers remain unchanged following one year of validated smoking cessation. AB - BACKGROUND: Elevated plasma concentrations of C-reactive protein (CRP) and total sialic acid (TSA) have been associated with increased cardiovascular risk. Additionally, levels of both CRP and TSA have been reported to be significantly elevated in smokers. However, it is not clear if the raised TSA and CRP levels noted in smokers are directly attributable to the smoking experience, or if they may be elevated due to a secondary mechanism(s), such as smoking-induced tissue inflammation. SUBJECTS AND METHODS: We measured the plasma concentration of CRP and TSA in a group of smokers at baseline and following one year of validated smoking cessation (n = 30) and in a control group of tobacco users who continued to smoke over the year (n = 30). RESULTS: The baseline concentration of TSA and CRP was 67.2 mg dL(-1) and 1.91 mg L(-1), respectively (n = 60). No significant dose-dependent relationship was noted between baseline CRP or TSA concentration and either plasma cotinine, expired-air CO or daily cigarette consumption. There was no difference in the mean change in CRP level in the quitters over one year ( 0.2 mg L(-1)) compared to the continuing smokers (+ 0.5 mg L(-1)), P = 0.80, or in the change in concentration of TSA in the quitters (- 2.7 mg dL(-1)) compared to the continuing smokers (+ 0.4 mg dL(-1)), P = 0.26. CONCLUSIONS: As the circulating concentrations of both CRP and TSA remain unchanged following one year of smoking cessation, these results would suggest that the elevated levels noted in smokers are not directly attributable to tobacco use and are more likely to be elevated due to a secondary process that is yet to be established. PMID- 11029600 TI - LDL-apheresis in patients with nephrotic syndrome: effects on serum albumin and urinary albumin excretion. AB - BACKGROUND: Hyperlipidemia is a common feature of the nephrotic syndrome (NS). From retrospective studies, it has been suggested that aggressive lipid-lowering with low-density lipoprotein apheresis (LDL-A) may not only improve dyslipidemia but also decrease urinary albumin excretion and increase serum levels of albumin in patients with focal segmental sclerosis. METHODS: Seven patients (6 males) aged 44 +/-7 years (SEM) with NS (duration 29+/-11 months) of diverse etiologies were investigated in a prospective study. A fixed protocol of LDL-A was designed for treatment twice-a-week for 3 weeks and then once a week for 7 weeks. The effects of LDL-A on lipid parameters (cholesterol, triglycerides, HDL, Lp(a), apo A-I, apo B) and renal parameters (iohexol clearance, serum albumin and 24-h urinary albumin excretion) were evaluated. RESULTS: Following treatment by LDL-A a remission in the severity of the NS was observed in two patients whereas a clear improvement was observed in four of the patients. A small, but significant (P<0.05), increase in serum albumin levels from 20+/-2 to 24+/-2 g L(-1) was noted after LDL-A. As expected, serum lipid parameters improved during LDL-A, and significant decreases in serum cholesterol, apo B and plasma Lp(a) were observed at different time-points of LDL-A. Conversely, no significant changes in either triglyceride, HDL or apo A-I levels were observed during LDL-A. CONCLUSIONS: The present uncontrolled prospective study shows that LDL-A causes a rapid 30-40% decrease in serum cholesterol and plasma Lp(a) levels in patients with NS. The present prospective study also suggests that short-term LDL-A treatment may increase serum albumin levels in nephrotic patients. PMID- 11029601 TI - Effects of comicronised fenofibrate on lipid and insulin sensitivity in patients with polymetabolic syndrome X. AB - BACKGROUND: This study investigated the effects of comicronised fenofibrate in patients with dyslipidemia and polymetabolic syndrome X. DESIGN: After a 6-week dietary run-in phase, 37 male patients eligible on lipid criteria entered a 12 week treatment phase consisting of diet plus one capsule daily containing 200 mg of comicronised fenofibrate (Lipanthyl(R)). RESULTS: A significant reduction in plasma concentrations of total cholesterol, LDL cholesterol and triglyceride was observed after 4, 8 and 12 weeks of treatment with fenofibrate. The improvement in the atherogenic index LDL/HDL cholesterol from a pretreatment 3.8 to 3.0 after treatment was highly statistically significant and may be judged as satisfactory. Significant changes were also observed in haemostatic factors (fibrinogen reduced by 19%, factor VII activity reduced by 18%). Fasting serum insulin levels and insulin response (area under the curve) after oral glucose load were significantly reduced by 26.8% and 18.7%, respectively, indicating an improvement of insulin sensitivity. Systolic and diastolic blood pressure were significantly reduced. Uric acid was significantly reduced by 21.6%. CONCLUSION: These favourable effects of comicronised fenofibrate both on lipid and non lipid parameters, including insulin sensitivity, may confer to this product a particular interest in the treatment of patients with polymetabolic syndrome X. PMID- 11029602 TI - Mutation analysis of the human adipocyte-specific apM-1 gene. AB - BACKGROUND: The aim of this study was to analyse the human adipocyte-specific apM 1 gene for sequence variations. METHODS: Sequence analysis was performed in 344 randomly chosen blood samples using a capillary sequencer. RESULTS: Whereas no mutations were detected in intronic regions and in 2.7 kb of the promoter, two sequence variations were found within the coding sequence of apM-1. For both mutations, a polymerase chain reaction-(PCR) based restriction fragment length polymorphism (RFLP) analysis was developed, which provided a rapid screening method. A conservative T --> G transition at nucleotide + 45 within exon-2 [Gly15Gly] was detected with an allelic frequency of 0.9 for the wild-type allele and 0.1 for the mutated allele. In addition, a missense point mutation at nucleotide + 331 within exon-3 [Tyr111His] was detected with an allelic frequency of 0.97 for the wild-type allele and 0.03 for the mutated allele. This mutation replaces a tyrosine by an histidine within the carboxyterminal globular domain of apM-1. Concerning the Gly15Gly polymorphism, the TT genotype was found in 275 subjects (79.9%), the TG genotype in 67 subjects (19.5%) and the GG genotype in 2 subjects (0.6%): one with maturity onset diabetes of young age (MODY-diabetes) and one with Lipoatrophic Diabetes Syndrome (LPDS). Concerning the Tyr111His polymorphism, the TT genotype was found in 328 subjects (95.4%), the TC genotype in 15 subjects (4.3%) and the CC genotype in 1 subject (0.3%). CONCLUSION: The existence of two yet unknown mutations within the apM-1 gene was demonstrated and RFLP analysis was established for rapid screening. Well defined cohorts of patients are necessary to determine the putative role of apM-1 gene mutations in the pathogenesis of metabolic disorders. PMID- 11029603 TI - Metabolic age modelling: the lesson from centenarians. AB - Evolutionary theories of ageing, and data emerging from cellular and molecular biology of ageing, suggested that animals and humans capable of reaching an age close to the extreme limit of the life span should be equipped with a very efficient network of anti-ageing mechanisms. Indeed several evidences have demonstrated that starting from young to very old subjects, ageing is associated with a progressive remodelling. Thus, a new paradigm, the remodelling theory of age, was proposed. This theory, focusing on the human immune system, suggested that immunosenescence is the net result of the continuous adaptation of the body to the deteriorative changes occurring over time. According to this hypothesis, body resources are continuously optimized, and immunosenescence must be considered a very dynamic process including both loss and gain. Whether the metabolic pathways and the endocrine functions are also part of the age remodelling is not investigated. The aim of this review is to focus on the age related changes in metabolic pathways and endocrine functions and to demonstrate that healthy centenarians (HC) represent the best living example of successful age-remodelling in whom the age remodelling has occurred without problems. In order to design the clinical picture of such successful ageing, anthropometric, endocrine and metabolic characteristics of healthy centenarians (HC), compared with aged subject, have been outlined. PMID- 11029604 TI - Influence of vitamin D administration on bone ultrasound measurements in patients on anticonvulsant therapy. AB - BACKGROUND: The objective of the study was to evaluate bone mass status (as measured by bone ultrasound) in patients on anticonvulsant therapy, and the influence that Vitamin D administration exerts over it. MATERIALS AND METHODS: We measured and compared the basal serum levels of 25(OH)D3, parathyroid hormone (PTH), and phalangeal bone ultrasound (Ad-SOS), in 30 adult patients who were taking anticonvulsant drugs, with a control group of similar age and sex. We then gave the patients a large oral dose of 3 mg (120.000 UI) of 25(OH)D3, and repeated the measurements after one month. RESULTS: Basal 25(OH)D3 and Ad-SOS values were significantly lower, and PTH values significantly higher (P< 0.0001 in all), in the patient group. The low Ad-SOS values for the patients were independent of the treatment, but directly related to basal 25(OH)D3 levels (r = 0.45, P<0.01). There was a negative association between PTH and 25(OH)D3 (r = 0.64, P<0.0001), and no correlation between PTH y Ad-SOS (r = -0.20, p NS). After ingestion of the large dose of the vitamin D, the patient group registered a significant (P<0.0001) increase in 25(OH)D3 levels, their Ad-SOS values increased (P<0.0001) nearly to the mean basal value of the control group, and PTH decreased significantly (P<0.0001). CONCLUSIONS: These findings justify the need to assure adequate vitamin D intake in patients being treated with anticonvulsants, independently of the treatment, age, sex, and activity status, in order to prevent osteomalacia. PMID- 11029605 TI - Serum concentration of 5-S-cysteinyldopa in patients with melanoma. AB - BACKGROUND: 5-S-cysteinyldopa (5-S-DC) is a precursor of melanin. Its serum and urinary level can reflect melanoma progression. In this study we examined the concentration changes of 5-S-CD in melanomas of different clinical stages and in patients with different symptoms of melanoma, during and after treatment. METHOD: Serum samples were taken from 252 melanoma patients on 765 occasions, from June 1996 to July 1998. Levels of 5-S-CD were determined by HPLC. RESULTS: The value of 5-S-CD in patients with primary melanoma and in patients without symptoms ranged around the normal level. There was a significant difference between the values of patients with or without symptoms. There was also a significant difference between the 5-S-CD values at clinical Stage I and Stage III, as well as at clinical Stage II and Stage III, respectively. Analysing the values of patients with symptoms we found a significant difference between the mean values of primary tumour and stage III, between values in lymph node metastasis and stage III, between values in lung metastasis and stage III. The tumour burden was found to correlate with a rising marker level. In 7% of the symptomatic patients that had a marker level under the upper limit, amelanotic primary tumour was detected. CONCLUSION: According to the high marker level in lung and liver metastases, the marker might be useful in monitoring both patients with disease free ocular melanomas, to detect liver metastasis and high-risk patients after surgical removal of the primary tumour to reveal lung metastases. PMID- 11029606 TI - Virological and immunological effects of antioxidant treatment in patients with HIV infection. AB - BACKGROUND: Intracellular oxidative stress in CD4+ lymphocytes due to disturbed glutathione homeostasis may lead to impaired lymphocyte functions and enhanced HIV replication in patients with HIV infection, especially in those with advanced immunodeficiency. The aim of the present study was to assess whether short-term, high-dose antioxidant treatment might have effects on immunological and virological parameters in patients with HIV infection. MATERIALS AND METHODS: In this pilot study, we examined virological and immunological effects of antioxidant combination treatment for 6 days with high doses of N-acetylcysteine (NAC) and vitamin C in 8 patients with HIV infection. The following were assayed before, during and after antioxidant treatment: HIV RNA plasma levels; numbers of CD4+, CD8+, and CD14+ leukocytes in blood; plasma thiols; intracellular glutathione redox status in CD4+ lymphocytes and CD14+ monocytes; lymphocyte proliferation; lymphocyte apoptosis and plasma levels of tumour necrosis factor (TNF)alpha; soluble TNF receptors and neopterin in plasma. RESULTS: No significant changes in HIV RNA plasma levels or CD4+ lymphocyte counts in blood were noted during antioxidant treatment in the patient group. However, in the 5 patients with the most advanced immunodeficiency (CD4+ lymphocyte counts < 200 x 106 L(-1)), a significant rise in CD4+ lymphocyte count, a reduction in HIV RNA plasma level of 0.8 log, an enhanced lymphocyte proliferation and an increased level of intracellular glutathione in CD4+ lymphocytes were found. No change in lymphocyte apoptosis was noted. CONCLUSIONS: Short-term, high-dose combination treatment with NAC and vitamin C in patients with HIV infection and advanced immunodeficiency lead to immunological and virological effects that might be of therapeutic value. PMID- 11029608 TI - Protein sgp 120 as a marker of an acquired angioedema. AB - BACKGROUND: Angioedema is not an event peculiar to an acute-phase response. We registered the appearance of angioedema in four nonatopic patients suffering from diverse inflammatory stimuli. MATERIALS AND METHODS: Serum concentrations of C reactive protein (CRP), haptoglobin, alpha1-antitrypsin, alpha2-macroglobulin, C3, C4, C1INH and protein sgp 120 (classified as a regulatory protein of the complement system) were assessed over 7 consecutive days, beginning with the onset of angioedema in these four patients. RESULTS: Instead of returning to the normal level, CRP peaked again on the 4th day (A-test, P<0.05) and declined significantly by the 7th day (A-test, P<0.05). Serum levels of sgp 120, C3c and C1INH increased gradually up to the 5th day but only sgp 120 was significantly raised in relation to the initial value (P< 0.05). CONCLUSION: In our patients, we registered four diverse profiles of the acute-phase proteins (Kruskal-Wallis test, P<0.05) probably due to the diverse nature of the pre-existing inflammatory stimuli. According to these results, the unexpected appearance of angioedema implies the contribution of sgp 120 as the main source of fragments with kinin like properties. PMID- 11029607 TI - N-acetylcysteine replenishes glutathione in HIV infection. AB - BACKGROUND: Glutathione (GSH) deficiency is common in HIV-infected individuals and is associated with impaired T cell function and impaired survival. N acetylcysteine (NAC) is used to replenish GSH that has been depleted by acetaminophen overdose. Studies here test oral administration of NAC for safe and effective GSH replenishment in HIV infection. DESIGN: Oral NAC administration in a randomized, 8-week double-blind, placebo-controlled trial followed by optional open-label drug for up to 24 weeks. SUBJECTS: HIV-infected, low GSH, CD4 T cells < 500 micro L(-1), no active opportunistic infections or other debilitation; n = 81. Study conducted prior to introduction of protease inhibitors. RESULTS: Whole blood GSH levels in NAC arm subjects significantly increased from 0.88 mM to 0.98 mM, bringing GSH levels in NAC-treated subjects to 89% of uninfected controls (P = 0.03). Baseline GSH levels in the placebo group (0.91) remained essentially the same during the 8 week placebo-controlled trial. T cell GSH, adjusted for CD4 T cell count and beta2-microglobulin levels, also increased in the NAC-treated subjects (P = 0.04). Adverse effects were minimal and not significantly associated with NAC ingestion. CONCLUSION: NAC treatment for 8 weeks safely replenishes whole blood GSH and T cell GSH in HIV-infected individuals. Thus, NAC offers useful adjunct therapy to increase protection against oxidative stress, improve immune system function and increase detoxification of acetaminophen and other drugs. These findings suggest that NAC therapy could be valuable in other clinical situations in which GSH deficiency or oxidative stress plays a role in disease pathology, e.g. rheumatoid arthritis, Parkinson's disease, hepatitis, liver cirrhosis, septic shock and diabetes. PMID- 11029609 TI - Mouse cytokine gene nucleotide sequence alignments, 2000. Part II. PMID- 11029610 TI - Further characterization of MHC haplotypes demonstrates conservation telomeric of HLA-A: update of the 4AOH and 10IHW cell panels. AB - Cell panels have been used extensively in studies of polymorphism and disease associations within the major histocompatibility complex (MHC), but the results from these panels require continuous updates with the increasing availability of novel data. We present here an updated table of the typings of the 10IHW and 4AOH panels. Local data included are HFE, HERV-K(C4) and six microsatellites telomeric of HLA-A. Typings for class I, MICA (PERB11.1), MICB (PERB11.2), XA, XB, LMP2 and 10 microsatellites reported by others have also been consolidated in this table. The tabulation shows that the length of conservation in the human MHC is even more extensive than previously thought. Human MHC ancestral haplotypes are inherited as a conserved region of genomic sequence spanning some 6-8 megabases from the HLA class II region and beyond the HLA class I region up to and including the HFE gene. Numerous examples of historical recombinations were also observed. PMID- 11029611 TI - Molecular cloning of the cDNAs encoding the feline B-lymphocyte activation antigen B7-1 (CD80) and B7-2 (CD86) homologues which interact with human CTLA4 Ig. AB - We cloned the cDNAs encoding the feline homologues of B-lymphocyte activation antigens B7-1 (CD80) and B7-2 (CD86). We expressed recombinant feline CD80 and CD86 molecules by the baculovirus expression system, and demonstrated their binding ability to human CTLA4-murine immunoglobulin fusion protein. PMID- 11029612 TI - Nomenclature for factors of the HLA system, update June 2000. PMID- 11029613 TI - Nomenclature for factors of the HLA system, update July 2000. PMID- 11029615 TI - Inhibition of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor expression reduces dopaminergic sprouting in the injured striatum. AB - After striatal injury, sprouting dopaminergic fibres grow towards and intimately surround wound macrophages which, together with microglia, express the dopaminergic neurotrophic factors glial cell line-derived neurotrophic factor (GDNF) and brain derived neurotrophic factor (BDNF). To evaluate the importance of these endogenously secreted neurotrophic factors in generating striatal peri wound dopaminergic sprouting, the peri-wound expression of BDNF or GDNF was inhibited by intrastriatal infusion of antisense oligonucleotides for 2 weeks in mice. Knock-down of both BDNF and GDNF mRNA and protein levels in the wounded striatum were confirmed by in situ hybridization and enzyme-linked immunosorbent assay, respectively. Dopamine transporter immunohisto-chemistry revealed that inhibition of either BDNF or GDNF expression resulted in a marked decrease in the intensity of peri-wound sprouting. Quantification of this effect using [H3] mazindol autoradiography confirmed that peri-wound sprouting was significantly reduced in mice receiving BDNF or GDNF antisense infusions whilst control infusions of buffered saline or sense oligonucleotides resulted in the pronounced peri-wound sprouting response normally associated with striatal injury. BDNF and GDNF thus appear to be important neurotrophic factors inducing dopaminergic sprouting after striatal injury. PMID- 11029614 TI - Expression of c-Met in developing rat hippocampus: evidence for HGF as a neurotrophic factor for calbindin D-expressing neurons. AB - Hepatocyte growth factor-scatter factor (HGF) is expressed in different parts of the nervous system, and has been shown to exhibit neurotrophic activity. Here we show that c-Met, the receptor for HGF, is expressed in developing rat hippocampus, with the highest levels during the first postnatal weeks. To study the function of HGF, hippocampal neurons were prepared from embryonic rats and treated with different HGF concentrations. In these cultures, HGF increased the number of neurons expressing the 28-kDa calcium-binding protein (calbindin D) in a dose-dependent manner. The effect of HGF was larger than that observed with either brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3), and cotreatment of the cultures with HGF and the neurotrophins was additive with respect to calbindin D neurons. Besides affecting the number of neurons, HGF significantly increased the degree of sprouting of calbindin D-positive neurons, suggesting an influence on neuronal maturation. BDNF and NT-3 stimulated neurite outgrowth of calbindin D neurons to a much smaller degree. In contrast to calbindin D neurons, HGF did not significantly increase the number of neurons immunoreactive with the neurotransmitter gamma-aminobutyric acid (GABA) in the hippocampal cultures. Immunohistochemical studies showed that c-Met-, calbindin D and HGF-immunoreactive cells are all present in the dentate gyrus and partly colocalize within neurons. These results show that HGF acts on calbindin D containing hippocampal neurons and increases their neurite outgrowth, suggesting that HGF plays an important role for the maturation and function of these neurons in the hippocampus. PMID- 11029616 TI - Caspase-3 is activated following axotomy of neonatal facial motoneurons and caspase-3 gene deletion delays axotomy-induced cell death in rodents. AB - In this report, we examined the possible functions of the cell death protease, caspase-3, in the axotomy-induced apoptosis of facial motoneurons in newborn rodents. Using in situ hybridization and Western blot, we found higher levels of caspase-3 mRNA and pro-caspase-3 protein expression in motoneurons of neonatal and 2-week-old rats than adult rats. Following facial motoneuron axotomy, caspase 3 mRNA and protein expression increased in motoneurons of both neonatal and adult rats. However, using an antibody directed to the activated form of the caspase-3 protease, we found that catalytically active caspase-3 was present only in axotomized neonatal motoneurons. As motoneurons in neonatal but not adult rodents are susceptible to axotomy-induced apoptosis, we hypothesized that caspase-3 may play a role in their demise. To determine the necessity of caspase-3 activation in axotomy-induced apoptosis, we counted the number of surviving motoneurons at 4 and 7 days following axotomy in wild type mice and caspase-3 gene-deleted mice. There were nearly three times more surviving motoneurons in caspase-3 gene deleted mice than in wild type mice at both 4 days (mean 1074 vs. 464, P<0.005) and 7 days (mean 469 vs. 190, P<0.005) following injury, indicating a slower rate of death. Examination of the dying motoneurons using TUNEL staining (for fragmented DNA) and bisbenzimide staining (for nuclear morphology) revealed incomplete nuclear condensation in caspase-3-deficient motoneurons. These results demonstrate that caspase-3 activation plays important roles in the rapid demise of axotomized neonatal motoneurons. PMID- 11029617 TI - Glucocorticoid enhancement of memory consolidation in the rat is blocked by muscarinic receptor antagonism in the basolateral amygdala. AB - Glucocorticoid-induced memory enhancement is known to depend on beta-adrenoceptor activation in the basolateral amygdala (BLA). Additionally, inactivation of muscarinic cholinergic receptors in the rat amygdala blocks memory enhancement induced by concurrent beta-adrenergic activation. Together, these findings suggest that glucocorticoid-induced modulation of memory consolidation requires cholinergic as well as adrenergic activation in the BLA. Two experiments investigated this issue. The first experiment examined whether blockade of muscarinic cholinergic receptors in the BLA with atropine alters the memory enhancing effects of the systemically administered glucocorticoid dexamethasone. Dexamethasone (0.3, 1.0 or 3.0 mg/kg, s.c.) administered to rats immediately after inhibitory avoidance training produced dose-dependent enhancement of 48-h retention. Concurrent bilateral infusions of the muscarinic cholinergic antagonist atropine (0.5 microg in 0.2 microL per side) into the BLA blocked the memory enhancement. The second experiment investigated whether the BLA is a locus of interaction between glucocorticoid and muscarinic activation. The specific glucocorticoid receptor (GR or type II) agonist RU 28362 (1.0, 3.0 or 10 ng) was infused into the BLA either alone or together with atropine immediately after training. The GR agonist produced dose-dependent memory enhancement and atropine blocked the memory enhancement. These findings indicate that muscarinic cholinergic activation within the BLA is critical for enabling glucocorticoid enhancement of memory consolidation and that enhancement of memory induced by GR activation in the BLA requires cholinergic activation within the BLA. PMID- 11029618 TI - Role of the GABA(A) receptor gamma2 subunit in the development of gonadotropin releasing hormone neurons in vivo. AB - We have employed transgenic mouse models to examine the functional significance of the gamma2 subunit of the GABA(A) (gamma-aminobutyric acid) receptor to the correct development of gonadotropin-releasing hormone (GnRH) neurons in vivo. In the first experiment, the expression of gamma2 subunit protein by the GnRH phenotype was determined using transgenic mice in which GnRH gene sequences direct the expression of the LacZ reporter to the nucleus of the GnRH neurons. This greatly facilitates the immunocytochemical identification of non-nuclear located antigens within GnRH neurons and revealed that approximately 25% of juvenile GnRH neurons were immunoreactive for the gamma2 subunit and that this increased to 40% in pubertal mice. In the second experiment, GnRH mRNA expression was examined in the brains of gamma2 subunit knockout mice (gamma2(0/0)) and their wild-type (gamma2+/+) littermates at embryonic day 15 and postnatal days (P) 0 and 11-16 using in situ hybridization. The distribution and numbers of cells expressing GnRH mRNA in gamma2+/+ and gamma2(0/0) mice were not found to differ at any age. However, the GnRH mRNA content of medial septal cells was significantly lower in gamma2(0/0) compared with gamma2+/+ mice at P11-16 (P<0.05) and the same trend was observed for preoptic area neurons. These results demonstrate that while the gamma2 subunit of the GABA(A) receptor is expressed by postnatal GnRH neurons, their embryonic development does not require a functional gamma2 subunit. In contrast, postnatal GnRH mRNA expression was found to be dependent upon signalling through the GABA(A) receptor. PMID- 11029619 TI - Late postnatal reorganization of GABA(A) receptor signalling in native GnRH neurons. AB - The molecular and cellular characteristics of the gonadotropin-releasing hormone (GnRH) neurons have been difficult to ascertain due to their scattered distribution within the basal forebrain. Using morphological criteria coupled with single cell RT-PCR postidentification, we have developed a method for investigating native GnRH neurons in the mouse brain and used it to examine the development of GABA(A) receptor signalling in this phenotype. Following the harvesting of the cytoplasmic contents of individual GnRH neurons, single cell multiplex RT-PCR experiments demonstrated that GABAA receptor alpha1-5, beta1-3 and gamma2 & 3 subunit transcripts were expressed by both neonatal (postnatal day 5) and juvenile (day 15-20) GnRH neurons in a heterogeneous manner. Following puberty, this profile was reduced to a predominant alpha1, alpha5, beta1, gamma2 subunit complement in rostral preoptic area GnRH neurons of the adult female. Whole-cell patch-clamp recordings revealed little difference between juvenile and adult GnRH neurons in their resting membrane potential and spontaneous firing rates. All GnRH neurons were found to be subjected to a tetrodotoxin-insensitive, tonic GABAergic barrage signalling through the GABA(A) receptor. However, marked heterogeneity in the sensitivity of individual juvenile GnRH neurons to GABA was revealed and, in parallel with the change in subunit mRNA profile, this was dramatically reduced in the reproductively competent adult GnRH neurons. These findings provide the first electrical and molecular characterization of the GnRH phenotype and demonstrate a novel pattern of late postnatal reorganization of native GABA(A) receptor gene expression and signalling in the GnRH neuronal population. PMID- 11029620 TI - Neuronal, astroglial and microglial cytokine expression after an excitotoxic lesion in the immature rat brain. AB - Cytokines are important intercellular messengers involved in neuron-glia interactions and in the microglial-astroglial crosstalk, modulating the glial response to brain injury and the lesion outcome. In this study, excitotoxic lesions were induced by the injection of N-methyl-D-aspartate in postnatal day 9 rats, and the cytokines interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), tumour necrosis factor alpha (TNFalpha) and transforming growth factor beta 1 (TGF-beta1) analysed by ELISA and/or immunohistochemistry. Moreover, cytokine expressing glial cells were identified by means of double labelling with glial fibrillary acidic protein or tomato lectin binding. Our results show that both neurons and glia were capable of cytokine expression following different patterns in the excitotoxically damaged area vs. the nondegenerating surrounding grey matter (SGM). Excitotoxically damaged neurons showed upregulation of IL-6 and downregulation of TNFalpha and TGF-beta1 before they degenerated. Moreover, in the SGM, an increased expression of neuronal IL-6, TNFalpha and TGF-beta1 was observed. A subpopulation of microglial cells, located in the SGM and showing IL 1beta and TNFalpha expression, were the earliest glial cells producing cytokines, at 2-10 h postinjection. Later on, cytokine-positive glial cells were found within the excitotoxically damaged area and the adjacent white matter: some reactive astrocytes expressed TNFalpha and IL-6, and microglia/macrophages showed mild IL-1beta and TGF-beta1. Finally, the expression of all cytokines was observed in the glial scar. As discussed, this pattern of cytokine production suggests their implication in the evolution of excitotoxic neuronal damage and the associated glial response. PMID- 11029621 TI - Developmental regulation of neurogenesis in the pluripotent human embryonal carcinoma cell line NTERA-2. AB - Embryonal carcinoma (EC) cells provide a caricature of pluripotent embryonic stem (ES) cells and may be used as surrogates for investigating the mechanisms that regulate cell differentiation during embryonic development. NTERA-2 is a human EC cell line that differentiates in response to retinoic acid yielding cells that include terminally differentiated neurons. The expression of genes known to be involved in the formation of the vertebrate nervous system was examined during retinoic acid-induced NTERA-2 differentiation. Differentiation of these human EC cells into neurons could be divided into three sequential phases. During phase 1, in the first week of differentiation, hath1 mRNA showed a small transient increase that correlated with the rapid accumulation of nestin message, a marker of neuroprogenitors. Transcripts of nestin were quickly downregulated during phase 2 as expression of neuroD1, characteristic of neuroprogenitors exiting the cell cycle, was induced. A neural cell surface antigen, detected by the monoclonal antibody A2B5, was expressed by cells exiting the cell cycle, correlating with the expression of neuroD1 as the cells became post-mitotic. Markers of mature neural cells (e.g. synaptophysin and neuron-specific enolase) were subsequently increased during phase 3 and were maintained. This regulated pattern of gene expression and commitment to the neural lineage indicates that differentiation of NTERA-2 neurons in vitro follows a similar pathway to that observed by neural ectodermal precursors during vertebrate neurogenesis in vivo. PMID- 11029622 TI - Inward current responses to urinary substances in rat vomeronasal sensory neurons. AB - No study has yet demonstrated an inward current in response to pheromonal substances in vomeronasal sensory neurons. Using female rat vomeronasal sensory neurons, we here successfully recorded inward currents in response to urine from various sources. Of the neurons that responded to urine, 77% responded to only one type of urine. Male Wistar urine induced responses preferentially in the apical layer of the sensory epithelium, whilst male Donryu and female Wistar urine induced responses mainly in the basal layer of the epithelium. The amplitude of inward currents induced by application of male Wistar urine was voltage-dependent with average amplitude of -47.1+/-6.2 pA at -74 mV. The average reversal potential for male Wistar urine was -9.3 +/-6.1 mV, which was not apparently different from the reversal potentials for urine from different species. It is likely that the urine-induced inward currents in response to different types of urine are mediated via a similar channel. The simultaneous removal of Na+ and Ca2+ from extracellular solution eliminated the response. The magnitude of the urine-induced inward current in Cl--free external solution was similar to that in normal solution, suggesting that the urine-induced current is cation selective. Removal of external Ca2+ enhanced the amplitude of the urine induced current and prolonged the response. Application of the constant-field equation indicated a very high permeability coefficient for Ca2+. This study first demonstrated that substances contained in urine elicited inward currents, which induce an excitatory response in vomeronasal sensory neurons, through cation-selective channels. PMID- 11029623 TI - Differential modulation of rod and cone calcium currents in tiger salamander retina by D2 dopamine receptors and cAMP. AB - Synaptic transmission from vertebrate photoreceptors involves activation of L type calcium currents (ICa). Dopamine is an important circadian neuromodulator in the retina and photoreceptors possess D2 dopamine receptors. We examined modulation of ICa by dopamine and cAMP in retinal slices and isolated cells of larval tiger salamander. Results show that dopamine and a D2 agonist, quinpirole, enhanced ICa in rods and red-, blue- and UV-sensitive small single cones but inhibited ICa in red-sensitive large single cones. A D1 agonist, SKF-38393, was without effect. Quinpirole effects were blocked by pertussis toxin (PTx) pretreatment indicating involvement of PTx-sensitive G-proteins. Like dopamine, inhibition of cAMP-dependent protein kinase (PKA) by Rp-cAMPS enhanced ICa in rods and small single cones, but inhibited ICa in large single cones. In contrast, forskolin and Sp-cAMPS, which stimulate PKA, inhibited ICa in rods and small single cones but enhanced ICa in large single cones. Sp-cAMPS also occluded effects of quinpirole. These results suggest that D2 receptors modulate ICa via inhibition of cAMP. Differences among the responses of photoreceptors to cAMP are consistent with the possibility that small single cones and rods may possess different Ca2+ channel subtypes than large single cones. The results with dopamine and quinpirole showing inhibition of ICa in large single cones and enhancement of rod ICa were unexpected because previous studies have shown that dopamine suppresses rod inputs and enhances cone inputs into second-order neurons. The present results therefore indicate that the dopaminergic enhancement of cone inputs does not arise from modulation of photoreceptor ICa. PMID- 11029624 TI - Excitable properties in astrocytes derived from human embryonic CNS stem cells. AB - Although it is widely believed that astrocytes lack excitability in adult tissue, primitive action potential-like responses have been elicited from holding potentials negative to -80 mV, in cultured and injury-induced gliotic rodent astrocytes and in human glia under pathological conditions such as glioblastomas and temporal lobe epilepsy. The present study was designed to investigate the properties of astrocytes (identified by immunoreactivity for glial fibrillary acidic protein) derived from multipotent human embryonic CNS stem cells and cultured for 12-25 days in differentiating conditions. We describe here for the first time that brief (1 ms) current pulses elicit spikes from a resting potential (VREST) of approximately -37 mV and, more interestingly, that spontaneous firing can be occasionally recorded in human astrocytes. A voltage clamp study revealed that in these cells: (i) the half-inactivation of the tetrodotoxin (TTX)-sensitive Na+ channels is around VREST; (ii) the delayed rectifier K+ current is very small; (iii) the ever-present transient outward A type K+ channels are paradoxically capable of inhibiting the action potentials elicited from a negative membrane potential (-55 to -60 mV); and (iv) inwardly rectifying currents are not present. The responses predicted from a simulation model are in agreement with the experiments. As suggested by recent studies, the decrease of Na+ channel expression and the changes of the electrophysiological properties during the postnatal maturation of the CNS seem to exclude the possibility that astrocytes may play an excitable role in adult tissue. Our data show that excitability and firing should be considered an intrinsic attribute of human astrocytes during CNS development. This is likely to have physiological importance because the role of astrocytes during development is different from the [K+]o-buffering role played in adult CNS, namely the glutamate release and/or the guiding of migrating neurons. PMID- 11029625 TI - Low-threshold heat response antagonized by capsazepine in chick sensory neurons, which are capsaicin-insensitive. AB - The heat-transducing receptor VR1 cloned from rat sensory neurons can be activated by both noxious heat and capsaicin. As the response of sensory neurons to capsaicin is species dependent, it is conceivable that the responses to noxious heat and to capsaicin are transduced by distinct receptors across different species. Therefore, we investigated responses to noxious heat from a capsaicin-insensitive (chick) and a capsaicin-sensitive (rat) species. In chick, whole-cell patch-clamp experiments in isolated dorsal root ganglion neurons revealed two populations of neurons with different thresholds to noxious heat, activated at approximately 43 degrees C and approximately 53 degrees C. In cobalt uptake experiments, the proportion of neurons showing a heat-induced response increased with increasing heat stimuli. Application of capsaicin (1-10 microM) did not result in inward currents or cobalt uptake. Rat neurons yielded comparable results in heat experiments, but were capsaicin-sensitive. Although chick neurons are insensitive to capsaicin, the competitive capsaicin antagonist capsazepine (1-10 microM) was effective in blocking heat-induced responses, verified by patch-clamp and cobalt uptake methods. The noncompetitive capsaicin antagonist ruthenium red (10 microM) reduced to almost nil the proportion of heat responsive neurons identified with the cobalt uptake method. These findings suggest that chick DRG neurons express a low-threshold heat-transducing receptor with a pharmacological profile distinct from the low-threshold heat receptor VR1 cloned from rat DRG neurons. The data support the idea that there might be heat receptor subtypes with differences in the capsaicin binding site. PMID- 11029626 TI - Riluzole inhibits the persistent sodium current in mammalian CNS neurons. AB - The effects of 0.1-100 microM riluzole, a neuroprotective agent with anticonvulsant properties, were studied on neurons from rat brain cortex. Patch clamp whole-cell recordings in voltage-clamp mode were performed on thin slices to examine the effects of the drug on a noninactivating (persistent) Na+ current (INa,p). INa,p was selected because it enhances neuronal excitability near firing threshold, which makes it a potential target for anticonvulsant drugs. When added to the external solution, riluzole dose-dependently inhibited INa,p up to a complete blocking of the current (EC50 2 microM), showing a significant effect at therapeutic drug concentrations. A comparative dose-effect study was carried out in the same cells for the other main known action of riluzole, the inhibitory effect on the fast transient sodium current. This effect was confirmed in our experiments, but we found that it was achieved at levels much higher than putative therapeutic concentrations. Only the effect on INa,p, and not that on fast sodium current, can account for the reduction in neuronal excitability observed in cortical neurons following riluzole treatment at therapeutic concentrations, and this might represent a novel mechanism accounting for the anticonvulsant and neuroprotective properties of riluzole. PMID- 11029627 TI - Expression of GABA(C) receptor rho1 and rho2 subunits during development of the mouse retina. AB - Retinal gamma-aminobutyric acid type C (GABA(C)) receptors consist of rho subunits. Here we report our results from a competitive PCR and patch-clamp electrophysiology study quantifying rho subunit message and characterizing GABA(C) receptor-mediated currents at different stages of mouse retinal development. Mouse rho1 message is first detected at postnatal day 6 (P6), increases significantly until P9 and remains at this level through adulthood, whereas mouse rho2 message does not appear until P9, peaks at P15 and remains at this level through adulthood. There is an approximate twofold excess of rho1 compared to rho2 message at most stages of development, which persists in adulthood. Functional GABA(C) receptors are detected in acutely dissociated bipolar cells of P9 or older mouse retina. Early in development (P9-10), GABA(C) receptors are composed solely of rho1 subunits, but subsequently contain rho1 and/or rho2 subunits (by P11 and later). These findings are intriguing because the onset and rapid increase in rho subunit transcription and functional expression match the initiation and active period of bipolar cell differentiation in retinal development as well as the stage of eye opening and initial visual experience in the rodent. The investigation of mouse rho subunits here forms a basis for future studies on the role of GABA(C) receptors in retinal development. PMID- 11029628 TI - Regional and cellular expression of the parkin gene in the rat cerebral cortex. AB - A mutation in the parkin gene has been identified as the cause for an autosomal recessively inherited form of early onset Parkinson's disease. We have recently isolated the mRNA coding for the rat homologue of parkin and showed its widespread expression in the central nervous system (CNS) by in situ hybridization. In the present study we investigated the distribution of parkin in the rat cerebral cortex with a polyclonal antibody that reacts with a single approximately 52-kDa protein, corresponding to the predicted molecular mass of parkin. Conventional light microscopic studies revealed intense parkin immunoreactivity (IR) throughout the cortex. Examination of mixed cortical neuro glial cultures by indirect immunofluorescence technique coupled to traditional epifluorescence and confocal microscopy analysis demonstrated the expression of parkin in the cytoplasm and neurites of neurons, and its absence in glial fibrillary acidic protein (GFAP)-positive astrocytes. The predominant neuronal parkin-IR and -mRNA expression was confirmed by Western blot analysis and reverse transcription-polymerase chain reaction (RT-PCR), respectively, performed on highly enriched neuronal and type I astrocytes cultures. The information gathered in our study about the cellular and subcellular distribution of parkin should facilitate further research on its physiological role in the nervous system. PMID- 11029629 TI - Functional connections are established in the deafferented rat spinal cord by peripherally transplanted human embryonic sensory neurons. AB - Functionally useful repair of the mature spinal cord following injury requires axon growth and the re-establishment of specific synaptic connections. We have shown previously that axons from peripherally grafted human embryonic dorsal root ganglion cells grow for long distances in adult host rat dorsal roots, traverse the interface between the peripheral and central nervous system, and enter the spinal cord to arborize in the dorsal horn. Here we show that these transplants mediate synaptic activity in the host spinal cord. Dorsal root ganglia from human embryonic donors were transplanted in place of native adult rat ganglia. Two to three months after transplantation the recipient rats were examined anatomically and physiologically. Human fibres labelled with a human-specific axon marker were distributed in superficial as well as deep laminae of the recipient rat spinal cord. About 36% of the grafted neurons were double labelled following injections of the fluorescent tracers MiniRuby into the sciatic and Fluoro-Gold into the lower lumbar spinal cord, indicating that some of the grafted neurons had grown processes into the spinal cord as well as towards the denervated peripheral targets. Electrophysiological recordings demonstrated that the transplanted human dorsal roots conducted impulses that evoked postsynaptic activity in dorsal horn neurons and polysynaptic reflexes in ipsilateral ventral roots. The time course of the synaptic activation indicated that the human fibres were non-myelinated or thinly myelinated. Our findings show that growing human sensory nerve fibres which enter the adult deafferentated rat spinal cord become anatomically and physiologically integrated into functional spinal circuits. PMID- 11029630 TI - Laterodorsal tegmental stimulation elicits dopamine efflux in the rat nucleus accumbens by activation of acetylcholine and glutamate receptors in the ventral tegmental area. AB - Cholinergic and glutamatergic neurons in the laterodorsal tegmentum (LDT) and neighbouring mesopontine nuclei are thought to influence mesolimbic dopaminergic neuronal activity involved in goal-directed behaviours. We measured the changes in dopamine oxidation current (corresponding with dopamine efflux) in the nucleus accumbens (NAc) in response to electrical stimulation of the LDT using in vivo chronoamperometry in urethane-anaesthetized rats. LDT stimulation (35 Hz pulse trains for 60 s, 1 s intertrain interval) evoked a three-component change in dopamine efflux in the NAc: (i) an initial stimulation time-locked increase in the dopamine signal above baseline, followed by (ii) an immediate decrease below baseline, and thereafter by (iii) a prolonged increase in the dopamine signal above baseline. Intra-VTA infusion of the nicotinic receptor antagonist mecamylamine (5 microg/0.5 microL) or the ionotropic glutamate receptor antagonist kynurenate (10 microg/microL) attenuated the first LDT-elicited component. The second suppressive component was abolished by intra-LDT infusions of either the nonselective or the M2-selective muscarinic receptor antagonists scopolamine (100 microg/microL) and methoctramine (50 microg/microL), respectively. In contrast, intra-VTA infusions of scopolamine (200 microg/microL) resulted in a selective attenuation of the third facilitatory component, whereas both second and third components were abolished by systemic injections of scopolamine (5 mg/kg). These results suggest that the initial increase, subsequent decrease, and final prolonged increase in extracellular dopamine levels in the NAc are selectively mediated by LDT-elicited activation of (i) nicotinic and glutamatergic receptors in the VTA, (ii) muscarinic M2 autoreceptors on LDT cell bodies, and (iii) muscarinic receptors in the VTA, respectively. PMID- 11029631 TI - SAP97 concentrates at the postsynaptic density in cerebral cortex. AB - SAP97, a PDZ-containing protein, is reported to concentrate in axon terminals, where its function remains unknown. Using highly specific new antibodies, we show that SAP97 in rat cerebral cortex is associated with heteromeric AMPA receptors via a selective biochemical interaction between SAP97 and the GluR1 subunit. Using light and electron microscopic immunocytochemistry, we demonstrate cellular and synaptic colocalization of SAP97 and GluR1, and show that SAP97 concentrates at synapses that contain GluR1 but not necessarily GluR2 or GluR3. Using quantitative postembedding immunogold electron microscopy, we find that SAP97 is at highest concentration within the postsynaptic density of asymmetric synapses. These data suggest that SAP97 may help to anchor GluR1-containing AMPA receptors at the synapse. As a multifunctional scaffolding protein, SAP97 may organize components of AMPA-related intracellular signalling pathways, including those associated with calcium-permeable homomeric GluR1 channels. PMID- 11029632 TI - Early and delayed induction of immediate early gene expression in a novel focal cerebral ischemia model in the rat. AB - This study aimed at evaluating changes in expression of immediate early genes in a new photothrombotic focal ischemia model that exhibits late spontaneous reperfusion and morphological restoration in the region-at-risk within the cerebral cortex. Gene expression was studied with Northern blots, in situ hybridization and immunohistochemistry. At early time points (1-4 h), nerve growth factor-induced gene A and B, and c-fos mRNAs, were quickly induced throughout the ipsilateral cortex, with no obvious differences between the region at-risk and remote cortical areas. High concentrations of nerve growth factor induced gene A and c-Fos proteins were present within the region-at-risk even when cortical cerebral blood flow was as low as 40% of control values. At 4 h the nerve growth factor-induced gene A mRNA and protein expression was significantly decreased in the hippocampus vs. naive controls. However, a small decrease was also found in sham-operated and anaesthetized controls. A late induction, at 5 days, of c-fos and nerve growth factor-induced gene B mRNAs was seen bilaterally in the hippocampus and also, in the case of nerve growth factor induced-gene B, in the contralateral cortex. A complex pattern of changes in immediate early gene expression occurs after reversible focal cortical ischemia. This may be important for tissue recovery as well as neuropsychiatric symptoms after stroke. PMID- 11029633 TI - Graded expression of EphA3 in the retina and ephrin-A2 in the superior colliculus during initial development of coarse topography in the wallaby retinocollicular projection. AB - We describe the expression of EphA3 and EphA7 receptors and ephrin-A2 ligand in the retina and the superior colliculus during the development of the retinocollicular projection in the marsupial wallaby (Macropus eugenii), using immunoblotting and immunohistochemistry. EphA3 in the retina was in a striking, low central to high peripheral gradient, superimposed on which was a high temporal to low nasal level of expression. This distribution was evident from postnatal day 30, when axons are growing into the colliculus and forming a coarsely organized topographic projection, to postnatal day 65, when axons have arborized in their correct retinotopic positions. EphA7 showed a shallow centroperipheral gradient with no nasotemporal differences in expression. In the superior colliculus no rostrocaudal differences in distribution were detected for either of these receptors. Ephrin-A2 was distributed in a gradient increasing from the rostral to the caudal pole in the superficial layers of the superior colliculus only up to postnatal day 30. Ephrin-A2 was evenly distributed in the retina throughout development of the projection. Expression of EphA3 in the retina increased, while the expression of ephrin-A2 in the colliculus was downregulated over time. The graded expression of EphA3 and ephrin-A2 early in the development of the projection suggests that they play a role in establishment of coarse topography of retinal axons along the rostrocaudal axis of the superior colliculus. However, the gradients were not complementary, meaning that EphA3 alone cannot mediate the repulsive interactions with ephrin-A2 that have been postulated to underlie formation of the topographic map. PMID- 11029634 TI - Tone-evoked oscillations in the rat auditory cortex result from interactions between the thalamus and reticular nucleus. AB - This study investigates the origins of tone-evoked oscillations (5-13 Hz) in the thalamo-cortical auditory system of anaesthetized rats. In three separate experiments, the auditory sector of the reticular nucleus (RE), the auditory cortex and the auditory thalamus were inactivated by local applications of muscimol (1 mg/mL). To assess the efficacy of this procedure, recordings were performed in the inactivated structure in each experiment; and to determine the extent of the drug diffusion autoradiographic experiments were carried out. The evolution of the strength of the oscillations was followed using power spectra during the whole recording session. In the first experiment, muscimol injection in the auditory RE totally suppressed the tone-evoked oscillations in the auditory thalamus and cortex. In the second experiment, inactivation of the auditory cortex did not interfere with the presence of tone-evoked oscillations in the auditory RE. In the third experiment, inactivation of the auditory thalamus impaired the oscillations produced by cortical stimulation in the auditory RE. From these results, it appears that both the auditory thalamus and the auditory sector of the RE, but not the auditory cortex, are involved in the generation of stimulus-evoked oscillations in the thalamo-cortical auditory system. PMID- 11029635 TI - Functional connections and epileptic spread between hippocampus, entorhinal cortex and amygdala in a modified horizontal slice preparation of the rat brain. AB - The hippocampus, the entorhinal cortex and the amygdala are interconnected structures of the limbic system that are implicated in memory and emotional behaviour. They demonstrate synaptic plasticity and are susceptible to development of temporal lobe epilepsy, which may lead to emotional and psychological disturbances. Their relative anatomical disposition has limited the study of neurotransmission and epileptic spread between these three regions in previous in vitro preparations. Here we describe a novel, modified-horizontal slice preparation that includes in the same plane the hippocampus, entorhinal cortex and amygdala. We found that, following application of bicuculline, each region in our preparation could generate spontaneous bursts that resembled epileptic interictal spikes. This spontaneous activity initiated in the hippocampal CA3/2 region, from where it propagated and controlled the activity in the entorhinal cortex and the amygdala. We found that this spontaneous bursting activity could spread via two different pathways. The first pathway comprises the well-known subiculum-entorhinal cortex-perirhinal cortex-amygdala route. The second pathway consists of a direct connection between the CA1 region and perirhinal cortex, through which the hippocampal bursting activity can spread to the amygdala while bypassing the entorhinal cortex. Thus, our experiments provide a new in vitro model of initiation and spread of epileptic-like activity in the ventral part of the limbic system, which includes a novel, fast and functional connection between the CA1 region and perirhinal cortex. PMID- 11029636 TI - Localization of nAChR subunit mRNAs in the brain of Macaca mulatta. AB - We present here a systematic mapping of nAChR subunit mRNAs in Macaca mulatta brain. A fragment, from the transmembrane segments MIII to MIV of Macaca neuronal nAChR subunits was cloned, and shown to exhibit high identity (around 95%) to the corresponding human subunits. Then, specific oligodeoxynucleotides were synthesized for in situ hybridization experiments. Both alpha4 and beta2 mRNA signals were widely distributed in the brain, being stronger in the thalamus and in the dopaminergic cells of the mesencephalon. Most brain nuclei displayed both alpha4 and beta2 signals with the exception of some basal ganglia regions and the reticular thalamic nucleus which were devoid of alpha4 signal. alpha6 and beta3 mRNA signals were selectively concentrated in the substantia nigra and the medial habenula. The strongest signals for alpha3 or beta4 mRNAs were found in the epithalamus (medial habenula and pineal gland), whereas there were no specific alpha3 or beta4 signals in mesencephalic dopaminergic nuclei. alpha5 and alpha7 mRNA signals were found in several brain areas, including cerebral cortex, thalamus and substantia nigra, although at a lower level than alpha4 and beta2. The distribution of alpha3, alpha4, alpha5, alpha6, alpha7, beta2, beta3 and beta4 subunit mRNAs in the monkey is substantially similar to that observed in rodent brain. Surprisingly, alpha2 mRNA signal was largely distributed in the Macaca brain, at levels comparable with those of alpha4 and beta2. This observation represents the main difference between rodent and Macaca subunit mRNA distribution and suggests that, besides alpha4beta2*, alpha2beta2* nAChRs constitute a main nAChR isoform in primate brain. PMID- 11029637 TI - Induction of VEGF and VEGF receptors in the spinal cord after mechanical spinal injury and prostaglandin administration. AB - Vascular endothelial growth factor (VEGF) is an angiogenetic factor that promotes endothelial cell proliferation during development and after injury to various types of tissue, including the central nervous system (CNS). Using immunohistochemical and in situ hybridization methods we have here demonstrated that VEGF and its receptors Flk-1, Flt-1 and Neuropilin-1 mRNAs and proteins are induced after incisions in the rat spinal cord. The inducible enzyme for prostaglandin synthesis cyclooxygenase-2 (COX-2) is known to be upregulated after spinal injury, cerebral ischemia and to stimulate angiogenesis. To test the hypothesis that prostaglandins may be involved in the VEGF response after lesion we investigated whether intraspinal microinjections of prostaglandin F2alpha (PGF2alpha) alters VEGF expression in the spinal cord. Such treatment was followed by a strong upregulation of VEGF mRNA and protein in the injection area. Finally, by use of an in vitro model with cell cultures of meningeal fibroblast and astrocyte origin, resembling the lesion area cellular content after spinal cord injury but devoid of inflammatory cells, we showed that VEGF is expressed in this in vitro model cell system after treatment with PGF2alpha and prostaglandin E2 (PGE2). These data suggest that cells within a lesion area in the spinal cord are capable of expressing VEGF and its receptors in response to mechanical injury and that prostaglandins may induce VEGF expression in such cells, even in the absence of inflammatory cells. PMID- 11029638 TI - Age-dependent pre- and postsynaptic distribution of AMPA receptors at synapses in CA3 stratum radiatum of hippocampal slice cultures compared with intact brain. AB - Organotypic slice cultures of rat hippocampus are widely used as experimental preparations for the study of synaptic plasticity, but their degree of correspondence with intact brain is not fully known. Here, using postembedding immunogold labelling, we describe the ultrastructural distribution of AMPA-type glutamate receptors (GluR1-4) in CA3 stratum radiatum of organotypic hippocampal slice cultures at 10 days to 11 weeks in vitro and compare the labelling with intact brain of corresponding age. In both types of preparation, the 11-week-old samples contained the highest proportion of AMPA receptor-like immunoreactive synapses. The incidence of labelled synapses, however, was higher in vivo (49%) than in vitro (24%). The intensity of labelling (number of gold particles per labelled synapse) also increased with age and was also higher in vivo than in vitro. In both organotypic cultures and intact brain, labelling was frequently found at presynaptic sites, often attached to vesicular structures. The specificity of these findings was supported both by light microscopic immunolabelling of GluR2/3 subunits and by electron microscopic double labelling of different epitopes of the GluR2 subunit. The vesicular localization of AMPA receptors was supported by Western blot analysis of subcellular fractions. Morphological evidence of presynaptic excitatory innervation of glutamatergic neurons supports a functional role for presynaptically located AMPA receptors. Our results therefore suggest that AMPA receptors occur in both pre- and postsynaptic profiles and that the distribution of AMPA receptors in cultured brain slices is fundamentally similar to intact brain, but that synaptic maturation may be retarded in vitro. PMID- 11029639 TI - Two splice variants of the hypoxia-inducible factor HIF-1alpha as potential dimerization partners of ARNT2 in neurons. AB - The hypoxia-inducible factor (HIF-1alpha), a basic helix-loop-helix transcription factor, is known to heterodimerize with ARNT1, a nuclear translocator, to trigger the overexpression in many cells of genes involved in resistance to hypoxia. Although HIF-1alpha and ARNT1 are both expressed in brain, their cellular localization and function therein are unknown. Here, using in situ hybridization and immunocytochemistry, we show that HIF-1alpha is expressed in normoxic cerebral neurons together with not only ARNT1 but also ARNT2, a cerebral translocator homologous to ARNT1 but displaying, unlike ARNT1, a selective neuronal expression. In contrast, other potential partners of the translocators, i.e. the aryl hydrocarbon receptor (AHR) and the single-minded protein 2 (SIM2), are not expressed in the adult brain. We also identify two splice variants of HIF 1alpha in brain, one of which dimerizes with ARNT2 even more avidly than with ARNT1. The resulting heterodimer, in contrast with the HIF-1alpha/ARNT1 complex, does not recognize the HIF-1-binding site of the hypoxia-induced erythropoietin (Epo) gene, suggesting that it controls transcription of a distinct set of genes. We therefore propose that HIF-1alpha and ARNT2 function as preferential dimerization partners in neurons to control specific responses, some of which may not be triggered by hypoxia. In support of this proposal, in nonhypoxic PC12 cells constitutively coexpressing HIF-1alpha, ARNT1 and ARNT2, downregulation of either HIF-1alpha or ARNT2, obtained with selective antisense nucleotides, resulted in inhibition of [3H]thymidine incorporation. PMID- 11029640 TI - Orbitofrontal involvement in the processing of unpleasant auditory information. AB - Although little is known about the contribution of the orbitofrontal cortex to the processing of new information in man, lesion studies in monkeys have suggested that it plays a critical role. The present study investigated changes in cerebral blood flow with positron emission tomography in normal human subjects during exposure to unpleasant auditory stimuli. The results indicated that the caudal orbitofrontal cortex, area 13, which is powerfully linked to the medial temporal limbic region and is involved in the regulation of autonomic responses, is a key part of the frontal cortex responding in the face of unpleasant incoming information. PMID- 11029641 TI - Acute transcranial magnetic stimulation of frontal brain regions selectively modulates the release of vasopressin, biogenic amines and amino acids in the rat brain. AB - Using intracerebral microdialysis in urethane-anaesthetized adult male Wistar rats, we monitored the effects of acute repetitive transcranial magnetic stimulation (rTMS; 20 trains of 20 Hz, 2.5 s) on the intrahypothalamic release of arginine vasopressin (AVP) and selected amino acids (glutamate, glutamine, aspartate, serine, arginine, taurine, gamma-aminobutyric acid) and the intrahippocampal release of monoamines (dopamine, noradrenaline, serotonin) and their metabolites (homovanillic acid, 3,4-dihydroxyphenylacetic acid, 5 hydroxyindoleacetic acid). The stimulation parameters were adjusted according to the results of accurate computer reconstructions of the current density distributions induced by rTMS in the rat and human brains, ensuring similar stimulation patterns in both cases. There was a continuous reduction in AVP release of up to 50% within the hypothalamic paraventricular nucleus in response to rTMS. In contrast, the release of taurine, aspartate and serine was selectively stimulated within this nucleus by rTMS. Furthermore, in the dorsal hippocampus the extracellular concentration of dopamine was elevated in response to rTMS. Taken together, these data provide the first in vivo evidence that acute rTMS of frontal brain regions has a differentiated modulatory effect on selected neurotransmitter/neuromodulator systems in distinct brain areas. PMID- 11029642 TI - Schizophrenia: glutathione deficit in cerebrospinal fluid and prefrontal cortex in vivo. AB - Schizophrenia is a major psychiatric disease, which affects the centre of the personality, with severe problems of perception, cognition as well as affective and social behaviour. In cerebrospinal fluid of drug-free schizophrenic patients, a significant decrease in the level of total glutathione (GSH) by 27% (P<0.05) was observed as compared to controls, in keeping with the reported reduced level of its metabolite gamma-glutamylglutamine. With a new non-invasive proton magnetic resonance spectroscopy methodology, GSH level in medial prefrontal cortex of schizophrenic patients was found to be 52% (P = 0.0012) lower than in controls. GSH plays a fundamental role in protecting cells from damage by reactive oxygen species generated among others by the metabolism of dopamine. A deficit in GSH would lead to degenerative processes in the surrounding of dopaminergic terminals resulting in loss of connectivity. GSH also potentiates the N-methyl-D-aspartate (NMDA) receptor response to glutamate, an effect presumably reduced by a GSH deficit, leading to a situation similar to the application of phencyclidine (PCP). Thus, a GSH hypothesis might integrate many established biological aspects of schizophrenia. PMID- 11029643 TI - Transient changes in cortical distribution of S100 proteins during reorganization of somatotopy in the primary motor cortex induced by facial nerve transection in adult rats. AB - In adult rats, the primary motor cortex (MI) comprises a somatotopic map of muscle representations. This somatotopy is modified after transection of the facial nerve (N7x). Mapping with cortical stimulation revealed that the underlying cortical reorganization is biphasic. Primary changes cause a transient disinhibition of long cortico-cortical connections in both hemispheres. While the first reaction vanishes within a few hours, short intra-areal connections are disinhibited within MI contralateral to N7x. The resulting co-operation between adjacent parts of MI persists as long as peripheral reinnervation is prevented. Cellular mechanisms underlying this cortical reorganization are largely unknown. Here, we utilized changes in immunoreactivity of S100 proteins (S100-IR) known as a sensitive indicator of astroglial reactions during plastic reactions in the central nervous system. Within 1 h of N7x, zones with enhanced S100-IR appeared in both hemispheres. Between 3. 5 and 18 h, reaction patterns with changing topography were transiently prominent in many cortical areas including parts of MI which surrounded the facial muscle representation fields. After 24 h, the facial muscle representation contralateral to N7x became labelled while S100-IR enhancement disappeared in most of the cortex. S100-IR-enhancement vanished completely during the next day of survival. Data presented suggest that (i) enhancement of S100-IR labels cortical tissue during the functional reorganization that is induced by N7x, (ii) large parts of the cerebral cortex participate in the reorganization, before it is finally focused on the representation field of MI that corresponds with contralateral N7x, and (iii) temporo-spatial patterns of astrocytic reactions apparently play a role in the underlying plasticity reaction. PMID- 11029644 TI - Activation of nicotinic acetylcholine receptors induces long-term potentiation in vivo in the intact mouse dentate gyrus. AB - The present study was conducted to clarify the role of nicotinic ACh receptors (nAChRs) on long-term potentiation (LTP) in vivo in the intact mouse dentate gyrus using extracellular recording techniques. Intraperitoneal application of nicotine at a dose of 3.0 mg/kg but not 0.03 or 0.3 mg/kg produced a gradually developing, long-lasting increase for 120 min similar to tetanic LTP. Nicotine at a dose of 9. 0 mg/kg caused a temporary increase followed by depression. The long lasting potentiation induced by nicotine at 3.0 mg/kg, which was named nicotinic long-term potentiation (LTPn), and tetanic LTP were significantly suppressed by pretreatment with mecamylamine (0.5 mg/kg i.p.), a nonselective nicotinic antagonist, but not affected by postapplication of mecamylamine. Interestingly, choline, a selective alpha7 nAChR agonist, at 3.0-90 mg/kg, induced the long lasting potentiation similar to LTPn in a dose-dependent manner in vivo in the intact mouse dentate gyrus. The long-lasting potentiation induced by choline (30 mg/kg i.p.) was additionally increased by postapplication of nicotine (3.0 mg/kg i.p.) or tetanic stimulation. The present study revealed that systemic application of nicotine or choline induced the long-lasting potentiation in vivo in the intact mouse dentate gyrus, suggesting that alpha7 nAChRs may contribute to the induction of LTP by nicotine, and supporting in vivo animal studies that nicotine improves learning and memory performance. PMID- 11029645 TI - Representation of accuracy in the dorsal premotor cortex. AB - The endpoint accuracy of a reaching movement strongly affects kinematics, particularly during the final phases of movement. However, where and how accuracy is represented in the central nervous system remains unknown. In this study, the discharge of 150 neurons located primarily in the dorsal premotor cortex (PMd), were recorded from monkeys performing an instructed delay, centre-out reaching task in which movement direction and target size were systematically varied. Linear regression analyses were used to assess the dependence of movement kinematics and cell discharge on target direction, size and tangential velocity (i.e. speed). The speed and timing of the movement were dependent on both direction and target size. Initially direction was the dominant predictor whilst target size became more important as the hand reached the target. A temporal multiple linear regression analysis found significant correlations with target size in 99 of 150 cells. The discharge of 134 cells was directionally tuned and 83 cells modulated with mean speed. Significant correlations of discharge with target size occurred throughout the task as did correlations with direction. However, correlations with direction preferentially occurred early in the task, prior to movement onset, whilst correlations with target size tended to occur late, well after movement onset. This temporal dependency of the firing in relationship to target direction and size mirrored that observed for the kinematics. We conclude that the discharge of PMd cells is highly correlated with the accuracy requirement of the movement. The timing of the correlations suggest that accuracy information is available for the planning and for the on-line control of endpoint accuracy. PMID- 11029646 TI - Involvement of sst2 somatostatin receptor in locomotor, exploratory activity and emotional reactivity in mice. AB - Somatostatin (SRIF) controls many physiological and pathological processes in the central nervous system but the respective roles of the five receptor isotypes (sst1-5) that mediate its effects are yet to be defined. In the present study, we attempted to identify functions of the sst2 receptor using mice with no functional copy of this gene (sst2 KO mice). In contrast with control 129Sv/C57Bl6 mice, sst2 mRNA was no longer detectable in the brain of sst2 KO mice; 125I-labeled Tyr0DTrp8-SRIF14 binding was also greatly reduced in almost all brain structures except for the hippocampal CA1 area, demonstrating that sst2 accounts for most SRIF binding in mouse brain. Invalidation of this subtype generated an increased anxiety-related behaviour in a number of behavioural paradigms, while locomotor and exploratory activity was decreased in stress inducing situations. No major motor defects could be detected. sst2 KO mice also displayed increased release of pituitary ACTH, a main regulator of the stress response. Thus, somatostatin, via sst2 receptor isotype pathways, appears involved in the modulation of locomotor, exploratory and emotional reactivity in mice. PMID- 11029647 TI - Maternal deprivation affects behaviour from youth to senescence: amplification of individual differences in spatial learning and memory in senescent Brown Norway rats. AB - Previous studies have shown that deprivation of the infant rat from maternal care has pronounced effects on the stress system during ontogeny. Here we test the hypothesis that 24 h of maternal deprivation at postnatal day 3 will cause persistent changes in behaviour. Spatial learning and memory of male Brown Norway rats deprived as infants were observed in the Morris water maze at 3, 12, 24 and 30-32 months of age (young, adult, aged, senescent). Their nondeprived mother reared littermates served as controls. (i) With increasing age, water maze performance declined in deprived and nondeprived groups. However, once the task was learned the animals maintained their good performance during retest at later ages. (ii) Maternal deprivation delayed acquisition until adulthood and caused at every age a higher degree of persistent behaviour as judged from the performance of deprived rats' free swim trials and reversal trials. (iii) At senescence the mean performance in the water maze did not differ between the groups. Instead, the individual performance was strikingly different within each group. Senescent deprived rats were either nonimpaired or impaired with only a few animals showing an intermediate performance. Thus, a large group of animals ( approximately 40%) ages successfully as they are resistant to the effect of maternal deprivation. In contrast, the majority of the control animals displayed intermediate performance. Taken together, maternal deprivation has life-long consequences for behaviour and culminates at senescence in amplification of individual differences in learning ability rather than in a generalized deterioration of cognitive functions. PMID- 11029648 TI - Experience-dependent plasticity of visual acuity in rats. AB - Rats have become a popular model for investigating the mechanisms underlying ocular dominance plasticity; however, no quantitative assessment of the effects of visual deprivation on behavioural acuity has been reported in this species. We measured the spatial acuity of monocularly and binocularly deprived rats with a visual discrimination task. The average spatial acuity of normal rats and rats deprived of vision after postnatal day 40 was approximately 1 cycle/degree. Monocular deprivation up to postnatal day 40 resulted in a 30% decrease in acuity and there was no recovery after 8 months. Identical binocular deprivation produced a comparable but significantly smaller reduction in acuity. The deleterious effects of monocular and binocular deprivation on visual acuity indicate that the development of cortical receptive field properties related to spatial tuning are affected by both monocular and binocular deprivation. The similarities in the effects of visual deprivation on visual acuity between rats and other mammals confirm that rats are a good model system for studying the cellular and molecular mechanisms underlying experience-dependent visual plasticity. PMID- 11029649 TI - Deficit in conditional visuomotor learning by local infusion of bicuculline into the ventral prefrontal cortex in monkeys. AB - To explore the role of the ventral prefrontal cortex (PFv) in conditional visuomotor learning, we infused locally bicuculline, a GABAergic antagonist, into the PFv of two monkeys, well trained on a two-problem visuomotor task. The task required the monkeys to execute one of two motor actions (moving a handle to the left or to the right) in response to one of two familiar visual patterns (circle or triangle). The two patterns mapped 1:1 onto the two motor actions: for each pattern one and only one motor action was scored, correct and reinforced. In contrast to these sessions with familiar patterns, in the learning sessions the monkeys were presented with one or two novel patterns and required to learn the arbitrarily determined associations between these patterns and the two motor actions. We found that bilateral infusion of bicuculline into PFv dramatically impaired the monkeys' ability to learn novel pattern-response associations: the trials and errors to criterion (90% correct) increased significantly. The errors were mainly an inability to apply 'Win-Stay', 'Lose-Shift' and 'Change-Shift' strategies. There was no effect on the monkeys' performance in responding to familiar patterns. Similar infusion of bicuculline into the dorsal prefrontal cortex was without effect on either novel-pattern learning or familiar-pattern performance. We conclude that the ventral prefrontal cortex is necessary for learning new visuomotor associations, but has less importance, if any, for performing pre-established ones. PMID- 11029650 TI - The role of 5-HT1A receptors in mediating the anxiogenic effects of nicotine following lateral septal administration. AB - The purpose of the present study was to determine the role of the 5-HT1A receptors in the lateral septum in the mediation of the anxiogenic effects of nicotine in the social interaction and elevated plus maze tests of anxiety in the rat. Bilateral infusion of (-)-nicotine (4 and 8 microg) and of the 5-HT1A receptor agonist 8-OH-DPAT (200 and 500 ng) into the lateral septum decreased the time spent in social interaction, indicating anxiogenic effects. The anxiogenic effect of 8-OH-DPAT (500 ng) was completely reversed by coadministration of a behaviourally inactive dose of the 5-HT1A receptor antagonist, WAY 100635 (200 ng). The anxiogenic effect of the lower dose of (-)-nicotine (4 microg) was completely reversed by WAY 100635 (200 ng), but the reversal was only partial following administration of 8 microg nicotine. In a second test of anxiety, the elevated plus maze, lateral septal administration of 8-OH-DPAT (500 ng) and nicotine (4 microg) induced anxiogenic effects. In this test, the anxiogenic effect of nicotine (4 microg) was completely reversed by coadministration of WAY 100635 (200 ng). The effects of 8-OH-DPAT demonstrate that stimulation of 5-HT1A receptors in the lateral septum has anxiogenic effects in two animal tests and that the anxiogenic effects of nicotine are mediated at least in part by these 5 HT1A receptors. PMID- 11029651 TI - Differential learning-stage dependent patterns of c-Fos protein expression in brain regions during the acquisition and memory consolidation of an operant task in mice. AB - The present study analysed the effects of the stage of learning of an appetitive operant conditioning task on the spatial and temporal patterns of c-Fos protein levels in the brain of BALB/c mice. c-Fos levels were assessed by immunohistochemistry at either 60, 120 or 180 min after either the first, the second or the fifth daily training session and compared to sham animals. The results show an increase of c-Fos-positive nuclei in several subcortical and cortical brain regions, 60-min post-acquisition. Because these activations were a function of task mastery, the data indicate that they were specifically related to learning. Following the first acquisition session, significant increases in c Fos-positive neurons were observed in the dorsal hippocampus (CA3), anterior cingulate, occipital and parietal cortices. Following the second daily training session, c-Fos was highly expressed in some subcortical regions, the hippocampus, the subiculum, the entorhinal, and posterior cingulate areas. Moreover, a significant correlation was found between the progression of performance from day 1 to day 2 and c-Fos expression on the hippocampal CA1 subfield. Following complete acquisition, no further task-dependent increases in c-Fos-labelled nuclei was observed in any brain region sampled, suggesting that the intervention of c-Fos-induced mechanisms in the consolidation process were terminated. The training stage-dependent changes in regional post-training c-Fos expression in the hippocampus and the connected limbic regions suggest that this neuronal network is actively engaged in memory consolidation processes. PMID- 11029652 TI - AMPA/kainate receptor-triggered Zn2+ entry into cortical neurons induces mitochondrial Zn2+ uptake and persistent mitochondrial dysfunction. AB - Rapid Zn2+ influx through Ca2+-permeable AMPA/kainate (Ca-A/K) channels triggers reactive oxygen species (ROS) generation and is potently neurotoxic. The first aim of this study was to determine whether these effects might result from direct mitochondrial Zn2+ uptake. Adapting the mitochondrially sequestered divalent cation sensitive probe, rhod-2, to visualize mitochondrial Zn2+, present studies indicate that Zn2+ is taken up into these organelles. The specificity of the signal for Zn2+ was indicated by its reversal by Zn2+ chelation, and its mitochondrial origin indicated by its speckled extranuclear appearance and by its elimination upon pretreatment with the mitochondrial protonophore, carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP). Consistent with inhibition of mitochondrial Zn2+ uptake, FCCP also slowed the recovery of cytosolic Zn2+ elevations in Ca-A/K(+) neurons. Further studies sought clues to the high toxic potency of intracellular Zn2+. In experiments using the mitochondrial membrane polarization (DeltaPsi(m))-sensitive probe tetramethylrhodamine ethyl ester and the ROS-sensitive probe hydroethidine, brief kainate exposures in the presence of 300 microM Zn2+ (with or without Ca2+) resulted in prolonged loss of DeltaPsi(m) and corresponding prolonged ROS generation in Ca-A/K(+) neurons, in comparison to the more rapid recovery from loss of DeltaPsi(m) and transient ROS generation after kainate/1.8 mM Ca2+ exposures. PMID- 11029653 TI - The spatial organization of cholinergic mosaics in the adult mouse retina. AB - We analysed the spatial organization of the cholinergic amacrine cell mosaics in the mouse retina, as part of a general study of the major mouse retinal arrays, aiming at providing intrinsic cellular reference grids to monitor anomalies in retinal growth and/or functional organization in mouse models of retinal degeneration. The spatial organization of the cells was analysed by means of the nearest neighbour distance analysis, as well as by the analysis of Voronoi and Delaunay tesselations. We found non random cell spacing in both cholinergic arrays, although the mosaic in the ganglion cell layer tiles the retina scarcely better than a random distribution. Autocorrelation analysis revealed no detectable pattern in cell positioning, but there was a tendency towards a minimal spacing between array elements. Finally, we found no correlation in the spatial organization of the two arrays. PMID- 11029654 TI - Nucleocytoplasmic protein traffic and its significance to cell function. AB - In eukaryotic cells, cell functions are maintained in an orderly manner through the continuous traffic of various proteins between the cell nucleus and the cytoplasm. The nuclear import and export of proteins occurs through nuclear pore complexes and typically requires specific signals: the nuclear localization signal and nuclear export signal, respectively. The transport pathways have been found to be highly divergent, but are known to be largely mediated by importin beta-like transport receptor family molecules. These receptor molecules bind to and carry their cargoes directly or via adapter molecules. A small GTPase Ran ensures the directionality of nuclear transport by regulating the interaction between the receptors and their cargoes through its GTP/GDP cycle. Moreover, it has been recently elucidated how the transport system is involved in various functions of cell physiology, such as cell cycle control. PMID- 11029655 TI - The absence of ribonuclease H1 or H2 alters the sensitivity of Saccharomyces cerevisiae to hydroxyurea, caffeine and ethyl methanesulphonate: implications for roles of RNases H in DNA replication and repair. AB - BACKGROUND: RNA of RNA-DNA hybrids can be degraded by ribonucleases H present in all organisms including the eukaryote Saccharomyces cerevisiae. Determination of the number and roles of the RNases H in eukaryotes is quite feasible in S. cerevisiae. RESULTS: Two S. cerevisiae RNases H, related to Escherichia coli RNase HI and HII, are not required for growth under normal conditions, yet, compared with wild-type cells, a double-deletion strain has an increased sensitivity to hydroxyurea (HU) and is hypersensitive to caffeine and ethyl methanesulphonate (EMS). In the absence of RNase H1, RNase H2 activity increases, and cells are sensitive to EMS but not HU and are more tolerant of caffeine; the latter requires RNase H2 activity. Cells missing only RNase H2 exhibit increased sensitive to HU and EMS but not caffeine CONCLUSIONS: Mutant phenotypes infer that some RNA-DNA hybrids are recognized by both RNases H1 and H2, while other hybrids appear to be recognized only by RNase H2. Undegraded RNA-DNA hybrids have an effect when DNA synthesis is impaired, DNA damage occurs or the cell cycle is perturbed by exposure to caffeine suggesting a role in DNA replication/repair that can be either beneficial or detrimental to cell viability. PMID- 11029656 TI - Two basic residues, Lys-107 and Lys-118, of RuvC resolvase are involved in critical contacts with the Holliday junction for its resolution. AB - BACKGROUND: Crystallographic and mutational studies of Escherichia coli RuvC Holliday junction resolvase have revealed that a catalytic site of each subunit is composed of four acidic residues at the bottom of the putative DNA-binding cleft, whose surface contains eight basic residues. RESULTS: To elucidate the functional roles of the basic residues on the cleft surface, we constructed a series of mutant ruvC genes and characterized their properties in vivo and in vitro. Among them, two RuvC mutants with a single alteration, K107A and K118A, were defective in UV-repair and showed a dominant negative effect. The purified K107A and K118A proteins showed reduced binding activity to the junction DNA in the presence of Mg2+ under high salt conditions. Mn2+ increased both the junction binding and cleaving activities of the mutant proteins. In the absence of a divalent cation, the wild-type, K107A and K118A proteins did not bind to junction DNA under high salt conditions, but the D7N mutant, with an alteration of the catalytic centre, was able to bind to the junction efficiently. CONCLUSION: The results presented here, in conjunction with previous crystallographic studies, suggest that the catalytic complex which is formed through interactions of acidic residues, Mg2+ and a cleavable phosphodiester bond, is stabilized by Lys-107 and Lys-118 via electrostatic interactions with the DNA backbone, a process which is critically important for the cleavage reaction to take place. One or two basic residues near the catalytic centre have also been found in other RNase H superfamily proteins, indicating that this is the conserved reaction mechanism in this superfamily. PMID- 11029657 TI - The colorectal tumour suppressor APC is present in the NMDA-receptor-PSD-95 complex in the brain. AB - BACKGROUND: The synaptic protein PSD-95/SAP90 interacts with ion channels such as the N-methyl-D-aspartate-receptor (NMDA-R) via its PDZ domain, and is involved in their clustering. Moreover, it interacts with signalling molecules and plays an important role in coupling NMDA-R to pathways that control synaptic plasticity and learning. RESULTS: We report that PSD-95 interacts with the adenomatous polyposis coli (APC) tumour suppressor protein via its PDZ domain. Furthermore, we found that PSD-95, NMDA-R and APC are contained in the same complex in vivo. PSD-95-NMDA-R-APC association was found to require two cysteine residues conserved in the amino-terminus of PSD-95 that are known to be critical for its multimerization. CONCLUSION: Our findings suggest that the PSD-95-NMDA-R-APC complex forms due to the multimerization of PSD-95 monomers, each of which can associate with either NMDA-R or APC. It is possible that APC is involved in the regulation of ion channel clustering and/or organization of signalling molecules. PMID- 11029658 TI - Localized phosphorylation of vimentin by rho-kinase in neuroblastoma N2a cells. AB - BACKGROUND: Vimentin, which is one of the intermediate filaments, is the major cytoskeletal component in developing neurones or neuroblastoma cells. Rho associated kinase (Rho-kinase), is rich in neurones and is found downstream of Rho. It is involved in the agonist-induced neurite retraction of neuronal cells, and phosphorylates vimentin at Ser-38 and Ser-71 resulting in in vitro disassembly of the filaments. RESULTS: We have investigated the distribution of vimentin phosphorylated by Rho-kinase in N2a neuroblastoma cells using site specific phosphorylation-dependent antibodies. TM71 immunoreactivity, which specifically indicates Ser-71 phosphorylation on vimentin, was found in some neurites of dibutyryl cAMP-differentiated N2a cells. Transfection of the constitutively active form of Rho-kinase, CAT, significantly elevated TM71 immunoreactivity, and induced neurite retraction or cell rounding. Conversely, transfection of the dominant negative form of Rho-kinase, RB/PH(TT), or treatment of 10 microM Y-27632, a Rho-kinase specific inhibitor, abolished TM71 immuno reactivity, and induced irregular neurite outgrowth. In contrast, 20 nM okadaic acid (OA) induced neurite retraction and specifically elevated TM71 immunoreactivity. In the OA-induced neurite retraction, tubulin disappeared in retracting neurites, where vimentin and actin remained co-localized. Furthermore, the OA-induced elevation of TM71 immunoreactivity and neurite retraction were completely blocked by pretreatment with 10 microM Y-27632, or by the ectopic expression of RB/PH(TT). CONCLUSIONS: This study suggests that the localized phosphorylation of vimentin by Rho-kinase in neurites was closely related with the cellular morphology of N2a cells, and that the Rho-kinase activity towards vimentin was balanced with OA-sensitive phosphatases. PMID- 11029659 TI - Identification and characterization of human Wee1B, a new member of the Wee1 family of Cdk-inhibitory kinases. AB - BACKGROUND: In eukaryotic cells, the kinase activity of the mitosis-promoting complex composed of cyclin B and Cdc2 (Cdk1) is negatively regulated by the phosphorylation of Cdk1 on threonine or tyrosine residues within its ATP binding domain. RESULTS: We identified human Wee1B by searching a sequence database. The predicted human Wee1B protein comprises 561 amino acids. Northern blot analysis revealed that human Wee1B mRNA is particularly abundant in testis. Interestingly, RT-PCR using early embryos revealed that the Wee1B product was readily detectable at the mature oocyte, but abruptly disappeared at embryonic day 2.5, suggesting that the amount of Wee1B mRNA is dependent on the maternal expression. GFP-Wee1B showed a predominantly nuclear localization in HeLa cells. Human Wee1B was able to rescue the lethal phenotype of the fission yeast wee1-50Deltamik1 mutant, and over-expression of the human protein in these cells resulted in cell elongation as a result of arrest of the cell cycle at the G2-M transition. Recombinant Wee1B effectively phosphorylated cyclin B-associated Cdk1 on tyrosine-15, resulting in an inactivation of the kinase activity of Cdk1. CONCLUSION: We identified human Wee1B as a novel Cdk1-inhibitory kinase. The identification of this new member of the Wee1 family suggests that inhibition of Cdk1 is mediated at multiple levels in mammals. PMID- 11029660 TI - LUCA15, a putative tumour suppressor gene encoding an RNA-binding nuclear protein, is down-regulated in ras-transformed Rat-1 cells. AB - BACKGROUND: The proliferation of mammalian cells is controlled by various intracellular mitogenic signalling pathways. In the intracellular pathways, Ras is involved in the activation of proto-oncogenes such as an immediate early gene c-fos. The somatic mutations of ras genes that elicit the constitutive activation of Ras have been found in tumours. Although these findings suggest that the constitutive activation of Ras-mediated pathways alters the expression of a set of genes involving tumorigenesis, these genes have not yet fully been studied. RESULTS: To study the up- or down-regulated genes in ras-transformed cells, we analysed Rat-1 transfectants expressing Ras(G12V) mutant protein in response to isopropyl-1-beta-thio-D-galactoside using a differential display. We found that the mRNA level of rat homologue of LUCA15, which has been cloned initially as a putative tumour suppressor gene mapped on human chromosome 3, was down-regulated by the expression of Ras(G12V). Epitope-tagged LUCA15 protein was localized in nuclei and had the ability to bind poly(G) RNA homopolymers in vitro. Moreover, ectopic expression of LUCA15 in human fibrosarcoma HT1080 cells suppressed the cell growth. CONCLUSION: These results demonstrate that LUCA15 is one of the down regulated genes in ras-transformed cells, and suggests that LUCA15 may function as a negative regulator of cell proliferation by the alteration of its mRNA level. PMID- 11029661 TI - Involvement of fatty acid synthase in axonal development in mouse embryos. AB - BACKGROUND: Dynamic protein palmitoylation of signalling proteins is thought to be an important step in the regulation of signal transduction in eukaryotic cells. The enzyme responsible for protein palmitoylation in vertebrates, however, has not been identified. I have previously reported that p260/270, which is expressed in embryos of the silkworm Bombyx mori, has protein palmitoylase activity. RESULTS: A homologue of Bombyx p260/270, mouse fatty-acid synthase (FAS), was shown to be expressed specifically throughout most of the central and peripheral nervous system in mouse embryos. Mouse FAS was expressed specifically in cultured primary neurones in which growth-associated protein (GAP)-43 was expressed. GAP-43, by protein palmitoylation, regulates Go signal transduction and neural axonal growth. In a cell-free system, purified FAS from mouse embryos transferred palmitate to GAP-43 through cysteine residues. Furthermore, cerulenin, an inhibitor of FAS, reduced axonal growth and in vivo palmitoylation of GAP-43 in cultured neurones. CONCLUSIONS: Mouse FAS was hypothesized to be responsible for the palmitoylation of GAP-43 and subsequent regulation of axonal growth in mouse embryos. PMID- 11029662 TI - Expression and activity of a house-fly cytochrome P450, CYP6D1, in Drosophila melanogaster. AB - The cytochrome P450 system of animals comprises many individual cytochromes P450 in addition to a single cytochrome P450 reductase and cytochrome b5. Although individual genes of the cytochrome P450 superfamily are highly diverged, the P450 reductase and cytochrome b(5) remain more conserved across taxa. Here, we describe the transformation of Drosophila melanogaster with a house-fly-specific cytochrome P450, CYP6D1. Functional activity of ectopically expressed cytochromes P450 requires successful interaction between the transgenic P450 and the requisite coenzymes of the host organism. Transformed Drosophila, but not controls, contained CYP6D1 protein as identified by protein immunoblotting, elevated total P450 and elevated CYP6D1 enzymatic activity. These data demonstrate that house-fly CYP6D1 can interact with low to moderate efficiency with Drosophila P450 reductase and cytochrome b(5). PMID- 11029663 TI - The kdr mutation occurs in the Mopti form of Anopheles gambiae s.s. through introgression. AB - Anopheles gambiae s.s. is a complex of sibling taxa characterized by various paracentric inversions. In west and central Africa, where several taxa are sympatric, a kdr mutation responsible for pyrethroid resistance has been described in only one (the S taxon), suggesting an absence of gene flow between them. Following a thorough sampling, we have found a kdr mutation in another taxon (M). To establish whether this mutation is the same event or not, the large intron upstream of the kdr mutation was sequenced to find polymorphic sites in susceptible/resistant and M/S mosquitoes. The low genetic diversity found in this DNA region indicates that a local genetic sweep has recently occurred. However, some polymorphic sites were found, and it is therefore concluded that the kdr mutation in the M taxon is not an independent mutation event, and is best explained by an introgression from the S taxon. These results are discussed within the context of possible gene flow between members of An. gambiae s.s. taxa, and with the possible spread of the kdr mutation in other closely related malaria vectors of the An. gambiae complex. PMID- 11029664 TI - Cloning and transcriptional expression of a leucokinin-like peptide receptor from the southern cattle tick, Boophilus microplus (Acari: Ixodidae). AB - Leucokinins are invertebrate neuropeptides that exhibit myotropic and diuretic activity. Only one leucokinin-like peptide receptor is known, the lymnokinin receptor from the mollusc Lymnaea stagnalis. A cDNA encoding a leucokinin-like peptide receptor was cloned from the Southern cattle tick, Boophilus microplus, a pest of cattle world-wide. This is the first neuropeptide receptor known from the Acari and the second known in the subfamily of leucokinin-like peptide G-protein coupled receptors. The deduced amino acid sequence exhibits 40% identity to the lymnokinin receptor. The receptor transcript is present in all tick life stages as determined by semiquantitative reverse transcription polymerase chain reaction. We also propose that the sequence AAF50775.1 from the Drosophila melanogaster genome (CG10626) encodes the first identified insect leucokinin receptor. PMID- 11029665 TI - Persistence of Anopheles arabiensis during the severe dry season conditions in Senegal: an indirect approach using microsatellite loci. AB - Variation at nine microsatellite loci was investigated to understand how Anopheles arabiensis populations survive the dry season in the sahelian region of Senegal. Low estimates of genetic differentiation (F(ST) = 0.012, R(ST) = 0.009) between two populations, 250 km apart, suggested extensive gene flow across this distance. Despite extreme seasonal fluctuation in abundance with dry season minima in which mosquitoes virtually disappeared, allele frequencies remained stable over time in the village of Barkedji from August 1994 to December 1997 (including four rainy seasons and three dry seasons). The effective population size (Ne) was estimated to be 601 with 95% CI (281, 1592), providing strong evidence against annual bottlenecks. Differences in measures of genetic diversity and linkage disequilibrium between the dry and the rainy seasons were not detected. These results suggest that despite extreme minima in local density, An. arabiensis maintains large permanent deme spread out over large area. PMID- 11029666 TI - Genomic organization and immune regulation of the defensin gene from the mosquito, Anopheles gambiae. AB - The defensin gene from the mosquito, Anopheles gambiae, is present as a single copy per haploid genome. Two exons, encoding a 102 residue preprodefensin, are separated by a 105 bp intron bounded by consensus splice sites. The upstream regulatory sequence includes a TATA box, arthropod initiator and numerous motifs homologous to insect and mammalian immune response elements. This promoter is capable of upregulation by immune challenge in cultured cells and activity is further stimulated by Gambif1, a mosquito Rel protein known to translocate to the nucleus and bind NF-kappa B sites in target promoters. Activity is inhibited by p50, a mammalian Rel protein that competitively binds NF-kappa B sites, and virtually abolished by p40, an avian I kappa B protein that inhibits nuclear translocation. PMID- 11029667 TI - Identification of a point mutation in the voltage-gated sodium channel gene of Kenyan Anopheles gambiae associated with resistance to DDT and pyrethroids. AB - A field trial of permethrin-impregnated bednets and curtains was initiated in Western Kenya in 1990, and a strain of Anopheles gambiae showing reduced susceptibility to permethrin was colonized from this site in 1992. A leucine phenylalanine substitution at position 1014 of the voltage-gated sodium channel is associated with resistance to permethrin and DDT in many insect species, including Anopheles gambiae from West Africa. We cloned and sequenced a partial sodium channel cDNA from the Kenyan permethrin-resistant strain and we identified an alternative substitution (leucine to serine) at the same position, which is linked to the inheritance of permethrin resistance in the F(2) progeny of genetic crosses between susceptible and resistant individuals. The diagnostic polymerase chain reaction (PCR) developed by Martinez-Torres et al. [(1998) Insect Mol Biol 7: 179-184] to detect kdr alleles in field populations of An. gambiae will not detect the Kenyan allele and hence reliance on this assay may lead to an underestimate of the prevalence of pyrethroid resistance in this species. We adapted the diagnostic PCR to detect the leucine-serine mutation and with this diagnostic we were able to demonstrate that this kdr allele was present in individuals collected from the Kenyan trial site in 1986, prior to the introduction of pyrethroid-impregnated bednets. The An. gambiae sodium channel was physically mapped to chromosome 2L, division 20C. This position corresponds to the location of a major quantitative trait locus determining resistance to permethrin in the Kenyan strain of An. gambiae. PMID- 11029668 TI - Genetic mapping of two loci affecting DDT resistance in the malaria vector Anopheles gambiae. AB - Resistance to the insecticide DDT in the mosquito vectors of malaria has severely hampered efforts to control this disease and has contributed to the increase in prevalence of malaria cases seen in recent years. Over 90% of the 300-500 million annual cases of malaria occur in Africa, where the major vector is Anopheles gambiae. DDT resistance in the ZAN/U strain of An. gambiae is associated with an increased metabolism of the insecticide, catalysed by members of the glutathione S-transferase (GST) enzyme family, but the molecular mechanism underlying this metabolic resistance is not known. Genetic crosses show that resistance is autosomal and semidominant. We have used microsatellite markers to identify two quantitative trait loci (QTL), which together explain over 50% of the variance in susceptibility to DDT in the ZAN/U strain of An. gambiae. The first locus, rtd1, is on chromosome 3 between markers H341 and H88 and has a recessive effect with respect to susceptibility. The second locus, rtd2 is on chromosome 2L, close to marker H325 and has an additive genetic effect. The markers flanking these two QTL have been physically mapped to An. gambiae polytene chromosomes. They do not coincide with any of the GST genes that have been cloned and mapped in this species. Characterization of these QTL will lead to a clearer understanding of the mechanisms of metabolic resistance to DDT. PMID- 11029669 TI - Identification of the sibling species of the Anopheles maculipennis complex by heteroduplex analysis. AB - The group of anopheline mosquitoes referred to as 'Anopheles maculipennis complex' includes the most important malaria vectors of the Palearctic Western region. The species belonging to this complex, however, are difficult or impossible to distinguish by morphological characters. To differentiate sibling palearctic species belonging to this complex, interspecific differences in the ITS2 sequences were used to set up a rapid and sensitive diagnostic tool based on heteroduplex analysis. The relative heteroduplex mobility allowed the following seven species to be readily distinguished: An. atroparvus, An. labranchiae, An. maculipennis s.s. , An. martinius, An melanoon, An. messeae and An. sacharovi. PMID- 11029670 TI - Subset partitioning of the ribosomal DNA small subunit and its effects on the phylogeny of the Anopheles punctulatus group. AB - A phylogenetic study, based on maximum parsimony, of ten species in the Anopheles punctulatus group of malaria vectors from the south-west Pacific was performed using structural and similarity-based DNA sequence alignments of the nuclear small ribosomal subunit (SSU = 18S). The structural alignment proved to be more informative than a computer generated similarity-based alignment. Analyses involving the full structural sequence alignment (2169 bp) and the helical regions (1547 bp) resolved a single tree of the same topology, while analyses using the similarity based alignment could not resolve the group. Studies on the three structural domains of the nuclear rDNA SSU identified domain 2 (769 bp) as the only region informative at the sibling-species level and resulted in the same tree as the full structural sequence and helical regions. The main conclusions of these studies were that the An. punctulatus group formed two clades: a Farauti clade containing members displaying an all black scaled proboscis (An. farauti 1 3 and 5-7) and a Punctulatus clade containing members that display some degree of white scaling on the proboscis (An. farauti 4, An. punctulatus and An. species near punctulatus). Anopheles koliensis can display either proboscis morphology and was positioned basal to the Farauti Clade. These results do not fully concord with those derived from the mitochondrial COII gene. PMID- 11029671 TI - The mitochondrial genome of the primary screwworm fly Cochliomyia hominivorax (Diptera: Calliphoridae). AB - The complete sequence of the mitochondrial genome of the screwworm Cochliomyia hominivorax was determined. This genome is 16,022 bp in size and corresponds to a typical Brachycera mtDNA. A Serine start codon for COI and incomplete termination codons for COII, NADH 5 and NADH 4 genes were described. The nucleotide composition of C. hominivorax mtDNA is 77% AT-rich, reflected in the predominance of AT-rich codons in protein-coding genes. Non-optimal codon usage was commonly observed in C. hominivorax mitochondrial genes. Phylogenetic analysis distributed the Acalypterate species as a monophyletic group and assembled the C. hominivorax (Calyptratae) and the Acalyptratae in a typical Brachycera cluster. The identification of diagnostic restriction sites on the sequenced mitochondrial genome and the correlation with previous RFLP analysis are discussed. PMID- 11029672 TI - Transformation of Stomoxys calcitrans with a Hermes gene vector. AB - The ability of the Hermes transposable element to function as a germ line transformation vector was tested in the stable fly, Stomoxys calcitrans. Plasmid based transposable element mobility assays indicated moderate mobility of Hermes in this species. Germline transformants were created using a Hermes element containing the enhanced green fluorescent protein (EGFP) under the regulatory control of the promoter from Actin5C gene of Drosophila melanogaster. Approximately 4% of the fifty-five adults that developed from the 1903 G(0) embryos injected with the vector produced transgenic progeny. In the four transgenic lines established, the EGFP expression pattern was distinctly nonuniform and levels of expression were low. Promoters other than the one from the Actin5C gene of D. melanogaster should be considered for widespread, constitutive expression. All transgenic lines contained multiple (2-4) integrated Hermes elements. Hermes integration events occurred through a canonical cut-and paste mechanism. PMID- 11029673 TI - Ultrastructural and molecular identification of a Wolbachia endosymbiont in a spider, Nephila clavata. AB - Wolbachia-like bacteria were observed in the egg cells of golden orb-weaving spider, Nephila clavata, by means of transmission electron microscopy. The bacteria exhibited the typical morphology of Wolbachia, including three enveloping membranes. Based on the amplification and sequencing of partial 16S rDNA and ftsZ gene, the bacteria were identified as Wolbachia, intracellular, transovarially inherited alpha-proteobacteria in invertebrates. Phylogenetic analysis based on 16S rDNA and ftsZ gene sequences invariably indicated that the intracellular bacteria from N. clavata belonged to group A Wolbachia, which were found only from insects. Clustering of Wolbachia from N. clavata with group A Wolbachia indicates that the bacteria were probably transferred horizontally between insects and the spider. PMID- 11029674 TI - Problems of older adults living alone after hospitalization. AB - OBJECTIVE: To describe functional deficits among older adults living alone and receiving home nursing following medical hospitalization, and the association of living alone with lack of functional improvement and nursing home utilization 1 month after hospitalization. DESIGN: Secondary analysis of a prospective cohort study. PARTICIPANTS: Consecutive sample of patients age 65 and over receiving home nursing following medical hospitalization. Patients were excluded for new diagnosis of myocardial infarction or stroke in the previous 2 months, diagnosis of dementia if living alone, or nonambulatory status. Of 613 patients invited to participate, 312 agreed. MEASUREMENTS: One week after hospitalization, patients were assessed in the home for demographic information, medications, cognition, and self-report of prehospital and current mobility and function in activities of daily living (ADLs) and independent activities of daily living (IADLs). One month later, patients were asked about current function and nursing home utilization. The outcomes were lack of improvement in ADL function and nursing home utilization 1 month after hospitalization. RESULTS: One hundred forty-one (45%) patients lived alone. After hospital discharge, 40% of those living alone and 62% of those living with others had at least 1 ADL dependency (P =.0001). Patients who were ADL-dependent and lived alone were 3.3 (95% confidence interval [95% CI], 1.4 to 7. 6) times less likely to improve in ADLs and 3.5 (95% CI, 1.0 to 11. 9) times more likely to be admitted to a nursing home in the month after hospitalization. CONCLUSION: Patients who live alone and receive home nursing after hospitalization are less likely to improve in function and more likely to be admitted to a nursing home, compared with those who live with others. More intensive resources may be required to continue community living and maximize independence. PMID- 11029675 TI - End-of-life decision making: a qualitative study of elderly individuals. AB - OBJECTIVE: To identify the desired features of end-of-life medical decision making from the perspective of elderly individuals. DESIGN: Qualitative study using in-depth interviews and analysis from a phenomenologic perspective. SETTING: A senior center and a multilevel retirement community in Los Angeles. PARTICIPANTS: Twenty-one elderly informants (mean age 83 years) representing a spectrum of functional status and prior experiences with end-of-life decision making. MAIN RESULTS: Informants were concerned primarily with the outcomes of serious illness rather than the medical interventions that might be used, and defined treatments as desirable to the extent they could return the patient to his or her valued life activities. Advanced age was a relevant consideration in decision making, guided by concerns about personal losses and the meaning of having lived a "full life." Decision-making authority was granted both to physicians (for their technical expertise) and family members (for their concern for the patient's interests), and shifted from physician to family as the patient's prognosis for functional recovery became grim. Expressions of care, both by patients and family members, were often important contributors to end-of life treatment decisions. CONCLUSIONS: These findings suggest that advance directives and physician-patient discussions that focus on acceptable health states and valued life activities may be better suited to patients' end-of-life care goals than those that focus on specific medical interventions, such as cardiopulmonary resuscitation. We propose a model of collaborative surrogate decision making by families and physicians that encourages physicians to assume responsibility for recommending treatment plans, including the provision or withholding of specific life-sustaining treatments, when such recommendations are consistent with patients' and families' goals for care. PMID- 11029676 TI - Communication breakdown in the outpatient referral process. AB - OBJECTIVE: To evaluate primary care and specialist physicians' satisfaction with interphysician communication and to identify the major problems in the current referral process. DESIGN: Surveys were mailed to providers to determine satisfaction with the referral process; then patient-specific surveys were e mailed to this group to obtain real-time referral information. SETTING: Academic tertiary care medical center. PARTICIPANTS: Attending-level primary care physicians (PCPs) and specialists. MEASUREMENTS AND MAIN RESULTS: The response rate for mail surveys for PCPs was 57% and for specialists was 51%. In the mail survey, 63% of PCPs and 35% of specialists were dissatisfied with the current referral process. Respondents felt that major problems with the current referral system were lack of timeliness of information and inadequate referral letter content. Information considered important by recipient groups was often not included in letters that were sent. The response rate for the referral specific e mail surveys was 56% for PCPs and 53% for specialists. In this e-mail survey, 68% of specialists reported that they received no information from the PCP prior to specific referral visits, and 38% of these said that this information would have been helpful. In addition, four weeks after specific referral visits, 25% of PCPs had still not received any information from specialists. CONCLUSIONS: Substantial problems were present in the referral process. The major issues were physician dissatisfaction, lack of timeliness, and inadequate content of interphysician communication. Information obtained from the general survey and referral-specific survey was congruent. Efforts to improve the referral system could improve both physician satisfaction and quality of patient care. PMID- 11029677 TI - Perceptions of benefit and risk of patients undergoing first-time elective percutaneous coronary revascularization. AB - OBJECTIVE: To assess reasons why patients undergo elective percutaneous coronary revascularization (PCR), patient expectations of the benefits of PCR, and their understanding of the risks associated with PCR. We hypothesized that patients overestimate the benefits and underestimate the risks associated with PCR. DESIGN: A prospective, semistructured questionnaire. PARTICIPANTS: Patients undergoing their first elective PCR. MAIN RESULTS: Fifty-two consecutive patients with a mean age of 64.3 years (range 39-87) completed the interview. Although 30 (57%) patients cited relief of symptoms as at least 1 reason to have PCR, 32 (62%) patients cited either an abnormal diagnostic test result (i.e., exercise stress test or catheterization) or "pathophysiologic" problem (i.e., "I have a blockage"), with 17 patients (33%) citing these reasons alone as indications for PCR. Thirty-nine (75%) patients believed PCR would prevent a future myocardial infarction, and 37 (71%) patients felt PCR would prolong their life. Regarding the potential complications, only 24 patients (46%) could recall at least 1 possible complication. However, on a Deber questionnaire, the majority of patients (67%) stated that they should determine either mostly alone or equally with a physician how acceptable the risks of the procedure are for themselves. CONCLUSIONS: The majority of patients had unrealistic expectations about the long term benefits of elective PCR and was not aware of the potential risks, even though they expressed a strong interest in participating in the decision to have PCR. More work is needed to define the optimal strategy to educate patients about the benefits and risks of elective PCR, and whether such education will affect patient decision making. PMID- 11029678 TI - Clinical presentation, processes and outcomes of care for patients with pneumococcal pneumonia. AB - OBJECTIVE: To describe the presentation, resolution of symptoms, processes of care, and outcomes of pneumococcal pneumonia, and to compare features of the bacteremic and nonbacteremic forms of this illness. DESIGN: A prospective cohort study. SETTING: Five medical institutions in 3 geographic locations. PARTICIPANTS: Inpatients and outpatients with community-acquired pneumonia (CAP). MEASUREMENTS: Sociodemographic characteristics, respiratory and nonrespiratory symptoms, and physical examination findings were obtained from interviews or chart review. Severity of illness was assessed using a validated prediction rule for short-term mortality in CAP. Pneumococcal pneumonia was categorized as bacteremic; nonbacteremic, pure etiology; or nonbacteremic, mixed etiology. MAIN RESULTS: One hundred fifty-eight (6.9%) of 2,287 patients (944 outpatients, 1,343 inpatients) with CAP had pneumococcal pneumonia. Sixty-five (41%) of the 158 with pneumococcal pneumonia were bacteremic; 74 (47%) were nonbacteremic with S. pneumoniae as sole pathogen; and 19 (12%) were nonbacteremic with S. pneumoniae as one of multiple pathogens. The pneumococcal bacteremia rate for outpatients was 2.6% and for inpatients it was 6.6%. Cough, dyspnea, and pleuritic pain were common respiratory symptoms. Hemoptysis occurred in 16% to 22% of the patients. A large number of nonrespiratory symptoms were noted. Bacteremic patients were less likely than nonbacteremic patients to have sputum production and myalgias (60% vs 82% and 33% vs 57%, respectively; P <.01 for both), more likely to have elevated blood urea nitrogen and serum creatinine levels, and more likely to receive penicillin therapy. Half of bacteremic patients were in the low risk category for short-term mortality (groups I to III), similar to the nonbacteremic patients. None of the 32 bacteremic patients in risk groups I to III died, while 7 of 23 (30%) in risk group V died. Intensive care unit admissions and pneumonia-related mortality were similar between bacteremic and nonbacteremic groups, although 46% of the bacteremic group had respiratory failure compared with 32% and 37% for the other groups. The nonbacteremic pure etiology patients returned to household activities faster than bacteremic patients. Symptoms frequently persisted at 30 days: cough (50%); dyspnea (53%); sputum production (48%); pleuritic pain (13%); and fatigue (63%). CONCLUSIONS: There were few differences in the presentation of bacteremic and nonbacteremic pneumococcal pneumonia. About half of bacteremic pneumococcal pneumonia patients were at low risk for mortality. Symptom resolution frequently was slow. PMID- 11029679 TI - Continuous quality improvement for patients with back pain. AB - Recent evidence has changed traditional approaches to low back pain, suggesting minimal bed rest, highly selective imaging, and early return to normal activities. However, there are wide geographical variations in care, and substantial gaps between practice and evidence. This project sought to merge scientific evidence about back pain and knowledge about behavior change to help organizations improve care for back pain. Participating insurance plans, HMOs, and group practices focused on problems they themselves identified. The year-long program included quarterly meetings, coaching for rapid cycles of change, a menu of potential interventions, and recommendations for monitoring outcomes. Participants interacted through meetings, e-mail, and conference calls. Of the 22 participating organizations, 6 (27%) made major progress. Typical changes were reduced imaging, bed rest, and work loss, and increased patient education and satisfaction. Specific examples were a 30% decrease in plain x-rays, a 100% increase in use of patient education materials, and an 81% drop in prescribed bed rest. Despite the complexity of care for back pain, rapid improvements appear feasible. Several organizations had major improvements, and most experienced at least modest improvements. Key elements of successful programs included focus on a small number of clinical goals, frequent measurement of outcomes among small samples of patients, vigilance in maintaining gains; involvement of office staffs as well as physicians, and changes in standard protocols for imaging, physical therapy, and referral. PMID- 11029680 TI - Factors influencing the selection of general internal medicine fellowship programs: a national survey. AB - Although criteria are available to guide the selection of general internal medicine (GIM) fellowship programs, the factors actually used in this process are unclear. Using a survey of current GIM fellows, we determined that most received information from their residency advisors, and many viewed them as the most important source of fellowship information. Program location was the top selection factor for fellows, followed by research opportunities, availability of a mentor, and the reputation of the program. This information may be useful to both fellowship candidates as an additional selection guide and to program directors seeking to best structure and market their fellowships. PMID- 11029681 TI - Treatment of fibromyalgia with antidepressants: a meta-analysis. AB - BACKGROUND: Fibromyalgia is a common, poorly understood musculoskeletal pain syndrome with limited therapeutic options. OBJECTIVE: To systematically review the efficacy of antidepressants in the treatment of fibromyalgia and examine whether this effect was independent of depression. DESIGN: Meta-analysis of English-language, randomized, placebo-controlled trials. Studies were obtained from searching MEDLINE, EMBASE, and PSYCLIT (1966-1999), the Cochrane Library, unpublished literature, and bibliographies. We performed independent duplicate review of each study for both inclusion and data extraction. MAIN RESULTS: Sixteen randomized, placebo-controlled trials were identified, of which 13 were appropriate for data extraction. There were 3 classes of antidepressants evaluated: tricyclics (9 trials), selective serotonin reuptake inhibitors (3 trials), and S-adenosylmethionine (2 trials). Overall, the quality of the studies was good (mean score 5.6, scale 0-8). The odds ratio for improvement with therapy was 4.2 (95% confidence interval [95% CI], 2.6 to 6.8). The pooled risk difference for these studies was 0.25 (95% CI, 0.16 to 0.34), which calculates to 4 (95% CI, 2.9 to 6.3) individuals needing treatment for 1 patient to experience symptom improvement. When the effect on individual symptoms was combined, antidepressants improved sleep, fatigue, pain, and well-being, but not trigger points. In the 5 studies where there was adequate assessment for an effect independent of depression, only 1 study found a correlation between symptom improvement and depression scores. Outcomes were not affected by class of agent or quality score using meta-regression. CONCLUSION: Antidepressants are efficacious in treating many of the symptoms of fibromyalgia. Patients were more than 4 times as likely to report overall improvement, and reported moderate reductions in individual symptoms, particularly pain. Whether this effect is independent of depression needs further study. PMID- 11029682 TI - Reporting by physicians of impaired drivers and potentially impaired drivers. The Committee on Bioethical Issues of the Medical Society of the State of New York. AB - Physicians routinely care for patients whose ability to operate a motor vehicle is compromised by a physical or cognitive condition. Physician management of this health information has ethical and legal implications. These concerns have been insufficiently addressed by professional organizations and public agencies. The legal status in the United States and Canada of reporting of impaired drivers is reviewed. The American Medical Association's position is detailed. Finally, the Bioethics Committee of the Medical Society of the State of New York proposes elements for an ethically defensible public response to this problem. PMID- 11029683 TI - Bringing a generalist approach to the problems of older patients. PMID- 11029684 TI - Two kinds of knowledge to achieve better care. PMID- 11029685 TI - A pot-pourri of plasmid paradoxes: effects of a second copy. AB - Bacterial plasmids are exemplary subjects for study, being conveniently isolated, dissected, reassembled, and introduced into various hosts. Their versatility and power make them eminently worthy of our attention. In what follows I consider some consequences of simply doubling the dosage of particular plasmid genes or of forming a plasmid dimer. These consequences can be perverse, paradoxical, or informative. They bear on questions of cell viability, copy number limitation, clonal homogeneity, check-point control, and the recovery of mutants. They have relevance to biotechnology, evolution and medicine. In reviewing these effects, my motivation is largely to share my enthusiasm for certain kinds of biological narratives, the nature of which is best left for the reader to discern. PMID- 11029686 TI - IpgD, a protein secreted by the type III secretion machinery of Shigella flexneri, is chaperoned by IpgE and implicated in entry focus formation. AB - Invasion of epithelial cells by Shigella flexneri involves entry and intercellular dissemination. Entry of bacteria into non-phagocytic cells requires the IpaA-D proteins that are secreted by the Mxi-Spa type III secretion machinery. Type III secretion systems are found in several Gram-negative pathogens and serve to inject bacterial effector proteins directly into the cytoplasm of host cells. In this study, we have analysed the IpgD protein of S. flexneri, the gene of which is located on the virulence plasmid at the 5' end of the mxi-spa locus. We have shown that IpgD (i) is stored in the bacterial cytoplasm in association with a specific chaperone, IpgE; (ii) is secreted by the Mxi-Spa type III secretion system in amounts similar to those of the IpaA-D proteins; (iii) is associated with IpaA in the extracellular medium; and (iv) is involved in the modulation of the host cell response after contact of the bacterium with epithelial cells. This suggests that IpgD is an effector that might be injected into host cells to manipulate cellular processes during infection. PMID- 11029687 TI - Regulation of carbon metabolism in Chlamydia trachomatis. AB - The biological significance of glycogen accumulation and how the process is regulated in Chlamydia trachomatis remains poorly defined. C. trachomatis infected HeLa cells were cultured in medium containing various glucose concentrations (0, 0.1, 1 or 10 mg ml-1) or in the presence of gluconeogenic carbon sources (20 mM glutamate, 20 mM malate, 20 mM alpha-ketoglutarate or 20 mM oxaloacetate), and the effects of these different culture conditions on the production of infectious chlamydial elementary bodies and glycogen accumulation were monitored. When chlamydiae were cultured in glucose concentrations greater than 1 mg ml-1, optimal growth and maximal glycogen accumulation occurred. In contrast to uninfected HeLa cells, which increased their glycogen stores when grown in the presence of high glucose concentrations, chlamydial glycogen accumulation remained essentially constant. When cultured in medium supplemented with either reduced glucose concentrations or any of the gluconeogenic carbon sources, chlamydiae still grew; however, the yield of elementary bodies was substantially decreased, and there was no significant amount of glycogen accumulated by host HeLa cells or C. trachomatis. This suggests that glycogen accumulation may not be essential for chlamydial survival. Reverse transcriptase polymerase chain reaction (RT-PCR) results indicated that, despite the fact that the source and amount of carbon available in the medium affected chlamydial glycogen accumulation, the expression of genes required for glycogen metabolism was not significantly changed. Similarly, the expression of several genes encoding key enzymes of central metabolism was not affected by alterations in carbon source or availability. Taken together, the data suggest that, unlike most free-living bacteria, chlamydia are unable to alter the expression of genes involved in carbon metabolism in response to changes in environmental conditions. PMID- 11029688 TI - Salmonella induces macrophage death by caspase-1-dependent necrosis. AB - We provide evidence that Salmonella typhimurium kills phagocytes by an unusual proinflammatory mechanism of necrosis that is distinguishable from apoptosis. Infection stimulated a distinctly diffuse pattern of DNA fragmentation in macrophages, which contrasted with the marked nuclear condensation displayed by control cells undergoing chemically induced apoptosis. In apoptotic cells, DNA fragmentation and nuclear condensation result from caspase-3-mediated proteolysis; caspases also subvert necrotic cell death by cleaving and inactivating poly ADP-ribose polymerase (PARP). Caspase-3 was not activated during Salmonella infection, and PARP remained in its active, uncleaved state. Another hallmark of apoptosis is sustained membrane integrity during cell death; yet, infected macrophages rapidly lost membrane integrity, as indicated by simultaneous exposure of phosphatidylserine with the uptake of vital dye and the release of the cytoplasmic enzyme lactate dehydrogenase. During experimentally induced necrosis, lethal ion fluxes through the plasma membrane can be prevented by exogenous glycine; similarly, glycine completely blocked Salmonella-induced cytotoxicity. Finally, inhibition of the interleukin (IL)-1-converting enzyme caspase-1 blocked the death of infected macrophages, but not control cells induced to undergo apoptosis or necrosis. Thus, Salmonella-infected macrophages are killed by an unusual caspase-1-dependent mechanism of necrosis. PMID- 11029689 TI - FlbT, the post-transcriptional regulator of flagellin synthesis in Caulobacter crescentus, interacts with the 5' untranslated region of flagellin mRNA. AB - Flagellar gene expression' in Caulobacter crescentus is regulated by a complex trans-acting hierarchy, in which the assembly of early structural proteins is required for the expression of later structural proteins. The flagellins that comprise the filament are regulated at both the transcriptional and the post transcriptional levels. Post-transcriptional regulation is sensitive to the assembly of the flagellar basal body and hook structures. In mutant strains lacking these structures, flagellin genes are transcribed, but not translated. Mutations in the flagellar regulatory gene, flbT, restore flagellin translation in the absence of flagellar assembly. In this report, we investigate the mechanism of FlbT-mediated post-transcriptional regulation. We show that FlbT is associated with the 5' untranslated region (UTR) of fljK (25 kDa flagellin) mRNA and that this association requires a predicted loop structure in the transcript. Mutations within this loop abolished FlbT association and resulted in increased mRNA stability, indicating that FlbT promotes the degradation of flagellin mRNA by associating with the 5' UTR. We also assayed the effects on gene expression using mutant transcripts fused to lacZ. Interestingly, the mutant transcript that failed to associate with FlbT in vitro was still repressed in mutants defective in flagellum assembly, suggesting that other factors in addition to FlbT couple assembly to translation. PMID- 11029690 TI - Identification of sat, an autotransporter toxin produced by uropathogenic Escherichia coli. AB - Urinary tract infection (UTI) is a very common extraintestinal infection, and Escherichia coli is by far the most common causative organism. Uropathogenic E. coli possess traits that distinguish them from commensal strains of E. coli, such as secretion systems that allow virulence factors to be targeted to extracytoplasmic compartments. One of at least five characterized secretion mechanisms is the autotransporter system, which involves translocation of a protein across the inner membrane, presumably via the sec system, and across the outer membrane through a beta-barrel porin structure formed by the carboxy terminus autotransporter domain. We identified a 107 kDa protein that was expressed significantly more often by E. coli strains associated with the clinical syndrome of acute pyelonephritis than by faecal strains (P = 0.029). We isolated the protein from E. coli CFT073, a strain cultured from the blood and urine of a patient with acute pyelonephritis. The N-terminal amino acid sequence showed highest similarity to two known SPATE (serine protease autotransporters of Enterobacteriaceae) proteins, Pet and EspC. Using a 509 bp probe from the 5' region of pet, 10 cosmid clones of an E. coli CFT073 gene library were positive for hybridization. From one cosmid clone, a 7.5 kb EcoRI restriction fragment, which reacted strongly with the probe, was shown to include the entire 3885 bp gene. The predicted 142 kDa protein product possesses the three domains that are typical of SPATE autotransporters: an unusually long signal sequence of 49 amino acids; a 107 kDa passenger domain containing a consensus serine protease active site (GDSGSG); and a C-terminal autotransporter domain of 30 kDa. The protein exhibited serine protease activity and displayed cytopathic activity on VERO primary kidney, HK-2 bladder and HEp-2 cell lines; the name Sat (secreted autotransporter toxin) was derived from these properties. In addition, Sat antibodies were present in the serum of mice infected with E. coli CFT073. Based upon its association with pathogenic isolates, its cytopathic phenotype and its ability to elicit a strong antibody response after infection, we postulate that Sat represents a novel virulence determinant of uropathogenic E. coli. PMID- 11029691 TI - The Vibrio cholerae ToxR/TcpP/ToxT virulence cascade: distinct roles for two membrane-localized transcriptional activators on a single promoter. AB - ToxR is required in Vibrio cholerae for transcriptional activation of the toxT gene, the protein product of which activates numerous genes involved in virulence. Although ToxR cannot activate the toxT promoter in Escherichia coli, the products of the tcpPH operon are shown here to activate the toxT promoter, and co-expression with ToxRS enhances activation. An identical pattern was seen in a DeltatcpPDeltatoxR strain of V. cholerae when TcpPH or ToxRS was expressed from plasmids. Although overexpression of the TcpP/H proteins in V. cholerae partially complemented both a DeltatoxR strain and a DeltatcpPDeltatoxR double mutant for toxin production and toxT-lacZ activation, the presence of ToxR greatly increased their expression. Analysis of a toxT-lacZ promoter deletion series demonstrated that TcpP was able to interact functionally with the toxT promoter downstream of the ToxR binding site. This was confirmed using electrophoretic mobility shift assays of this toxT promoter deletion series and DNase I footprinting analysis, which showed that TcpP interacts with the promoter region from -51 to -32, whereas ToxR protected a region from -100 to -69. In addition, membranes containing endogenous levels of ToxR bound more readily to the toxT promoter than did membranes containing only TcpP. Characterization of a number of tcpP substitution mutants revealed one derivative (TcpP-H93L) that, when overexpressed, was markedly defective for toxT activation, cholera toxin and TcpA (toxin co-regulated pilus) production and DNA binding; however, toxT activation by TcpP-H93L was restored in the presence of ToxR, suggesting that ToxR can provide the promoter recognition function for toxT activation. Two additional mutant derivatives, TcpP-W68L and TcpP-R86A, failed to activate toxT or direct toxin and TcpA production in the presence or absence of ToxR. Both TcpP W68L and TcpP-R86A, like TcpP-H93L, were defective for DNA binding. Finally, a ToxR mutant derivative, ToxR-G80S, served to separate the different roles of ToxR on different promoters. Although ToxR-G80S was inefficient at activating the ompU promoter in V. cholerae (ompU encodes an outer membrane porin regulated by ToxR), it was fully capable of activating the toxT promoter. These data suggest that ToxR is not a direct activator in the toxT expression system but, instead, enhances the activity of TcpP, perhaps by recruiting it to the toxT promoter under conditions in which expression levels of TcpP are too low for it to activate toxT efficiently on its own. PMID- 11029692 TI - A study of the YopD-lcrH interaction from Yersinia pseudotuberculosis reveals a role for hydrophobic residues within the amphipathic domain of YopD. AB - The enteropathogen Yersinia pseudotuberculosis is a model system used to study the molecular mechanisms by which Gram-negative pathogens translocate effector proteins into target eukaryotic cells by a common type III secretion machine. Of the numerous proteins produced by Y. pseudotuberculosis that act in concert to establish an infection, YopD (Yersinia outer protein D) is a crucial component essential for yop regulation and Yop effector translocation. In this study, we describe the mechanisms by which YopD functions to control these processes. With the aid of the yeast two-hybrid system, we investigated the interaction between YopD and the cognate chaperone LcrH. We confirmed that non-secreted LcrH is necessary for YopD stabilization before secretion, presumably by forming a complex with YopD in the bacterial cytoplasm. At least in yeast, this complex depends upon the N-terminal domain and a C-terminal amphipathic alpha-helical domain of YopD. Introduction of amino acid substitutions within the hydrophobic side of the amphipathic alpha-helix abolished the YopD-LcrH interaction, indicating that hydrophobic, as opposed to electrostatic, forces of attraction are important for this process. Suppressor mutations isolated within LcrH could compensate for defects in the amphipathic domain of YopD to restore binding. Isolation of LcrH mutants unable to interact with wild-type YopD revealed no single domain responsible for YopD binding. The YopD and LcrH mutants generated in this study will be relevant tools for understanding YopD function during a Yersinia infection. PMID- 11029693 TI - Helicobacter pylori induces but survives the extracellular release of oxygen radicals from professional phagocytes using its catalase activity. AB - Helicobacter pylori can colonize the gastric epithelium of humans, leading to the induction of an intense inflammatory response with the infiltration of mainly polymorphonuclear leucocytes (PMNs) and monocytes. These professional phagocytes appear to be a primary cause of the damage to surface epithelial layers, and probably contribute to the pathogenesis associated with persistent H. pylori infections. We have shown previously that H. pylori adheres to professional phagocytes, but is not engulfed efficiently, suggesting an antiphagocytic escape mechanism that is dependent on the pathogen's type IV secretion system. Here, we show that H. pylori induces the generation and extracellular release of oxygen metabolites as a consequence of its attachment to phagocytic cells, but is capable of surviving this response. The catalase activity of H. pylori is apparently essential for survival at the phagocytes' cell surface. Opsonization of H. pylori leads to an increased burst, and the inhibition of bacterial protein synthesis to a decreased one. Ca2+ concentration, cytoskeleton rearrangement and protein kinase C (PKC) are involved in the H. pylori-induced oxidative burst in both monocytes and PMNs. This survival phenomenon has important implications for both the persistence of this important pathogen and the host tissue damage that accompanies persistent H. pylori infection. PMID- 11029694 TI - Comparison of the sequences of the Aspergillus nidulans hxB and Drosophila melanogaster ma-l genes with nifS from Azotobacter vinelandii suggests a mechanism for the insertion of the terminal sulphur atom in the molybdopterin cofactor. AB - The molybdopterin cofactor (MoCF) is required for the activity of a variety of oxidoreductases. The xanthine oxidase class of molybdoenzymes requires the MoCF to have a terminal, cyanolysable sulphur ligand. In the sulphite oxidase/nitrate reductase class, an oxygen is present in the same position. Mutations in both the ma-l gene of Drosophila melanogaster and the hxB gene of Aspergillus nidulans result in loss of activities of all molybdoenzymes that necessitate a cyanolysable sulphur in the active centre. The ma-l and hxB genes encode highly similar proteins containing domains common to pyridoxal phosphate-dependent cysteine transulphurases, including the cofactor binding site and a conserved cysteine, which is the putative sulphur donor. Key similarities were found with NifS, the enzyme involved in the generation of the iron-sulphur centres in nitrogenase. These similarities suggest an analogous mechanism for the generation of the terminal molybdenum-bound sulphur ligand. We have identified putative homologues of these genes in a variety of organisms, including humans. The human homologue is located in chromosome 18.q12. PMID- 11029695 TI - A role for the Escherichia coli H-NS-like protein StpA in OmpF porin expression through modulation of micF RNA stability. AB - When a wild-type strain of Escherichia coli and its stpA, hns and stpA hns mutant derivatives were compared by two-dimensional protein gel electrophoresis, the levels of expression of several proteins were found to vary. One of these was identified as the outer membrane porin protein, OmpF. In the stpA hns double mutant, the level of OmpF was downregulated dramatically, whereas in hns or stpA single mutants, it was affected only slightly. Transcription from the ompF promoter was reduced by 64% in the double mutant; however, the level of ompF mRNA was reduced by 96%. This post-transcriptional expression was found to result from a strong reduction in the half-life of ompF message in the double mutant. The micF antisense RNA was shown to be involved in OmpF regulation by StpA using a strain deleted for micF. Moreover, micF antisense RNA accumulated considerably in an stpA hns background. Transcriptional data from a micF-lacZ fusion and measurements of micF RNA half-life confirmed that this was caused by transcriptional derepression of micF as a result of the hns lesion and increased micF RNA stability due to the absence of StpA (a known RNA chaperone). These data suggest a novel facet to the regulation of OmpF expression, namely destabilization of micF RNA by StpA. PMID- 11029696 TI - Expression of the virulence-related Sca (Mn2+) permease in Streptococcus gordonii is regulated by a diphtheria toxin metallorepressor-like protein ScaR. AB - The acquisition of transition metal ions by pathogenic bacteria is crucial to their growth and survival within the human host, however, the mechanisms of metal ion homeostasis in streptococci are unknown. The scaCBA operon in the human oral bacterium Streptococcus gordonii encodes the components of an ABC-type transporter for manganese (Mn2+). Production of substrate-binding lipoprotein ScaA was increased approximately fivefold in cells cultured in low Mn2+ medium (< 0.1 microM Mn2+), but not in iron (Fe2+/Fe3+)-limited medium, and was enhanced in the presence of human saliva or serum. mRNA analysis revealed that under low Mn2+ conditions, levels of scaCBA transcript (2.6 kb) were increased > 20-fold. The Mn2+-responsive transcriptional regulator of the sca operon was purified and characterized as a 215-amino-acid residue polypeptide, designated ScaR, with 26% identity to the Corynebacterium diphtheriae diphtheria toxin repressor (DtxR). Inactivation of scaR in S. gordonii DL1 (Challis) resulted in constitutive derepression of sca operon transcription. Expression of tpx, located immediately downstream of scaA and encoding a putative thiol peroxidase, was not subject to ScaR regulation. Purified ScaR protein bound to the scaC promoter region in vitro in the presence of Mn2+ (Kd approximately 80 nM) and, to a lesser extent, in the presence of Ni2+ or Zn2+. The metalloregulator protein binding region was localized by DNA protection analysis to a 46 bp sequence encompassing the -35 and -10 promoter signatures. This sequence was well conserved within the promoters of corresponding virulence-related permease operons in other streptococci. The results identify a new Mn2+-sensing regulator of Mn2+ transport in streptococci, important for Mn2+ homeostasis during infection of the human host. PMID- 11029697 TI - A protein with similarity to the human retinoblastoma binding protein 2 acts specifically as a repressor for genes regulated by the b mating type locus in Ustilago maydis. AB - Pathogenic development in the corn smut fungus Ustilago maydis is controlled by a heterodimer of the two homeodomain proteins bE and bW which are encoded by the b mating type locus. The bE/bW heterodimer is thought to achieve its function as a transcriptional regulator of pathogenicity genes, either directly by binding to cis regulatory sequences or indirectly via a b-dependent regulatory cascade. In a screen for components of the b-dependent regulatory cascade we have isolated Rum1 (regulator U. maydis 1), a protein with similarities to the human retinoblastoma binding protein 2. Deletion of rum1 results in expression of several b regulated genes independently from their activation via the bE/bW heterodimer. rum1 mutant strains remain pathogenic, proliferate in planta, but fail to produce spores. The defect leads to an arrest in spore development at a defined stage before the spore wall is generated. Deduced from the highly conserved domain structure of Rum1 that includes a DNA-binding motif and a region known to facilitate the interaction with histone deacetylases, we propose that Rum1 functions as a transcriptional repressor through the modulation of chromatin structure. PMID- 11029698 TI - The expression of the Escherichia coli fis gene is strongly dependent on the superhelical density of DNA. AB - The Escherichia coli DNA architectural protein FIS is a pleiotropic regulator, which couples the cellular physiology with transitions in the superhelical density of bacterial DNA. Recently, we have shown that this effect is in part mediated via DNA gyrase, the major cellular topoisomerase responsible for the elevation of negative supercoiling. Here, we demonstrate that, in turn, the expression of the fis gene strongly responds to alterations in the topology of DNA in vivo, being maximal at high levels of negative supercoiling. Any deviations from these optimal levels decrease fis promoter activity. This strict dependence of fis expression on the superhelical density suggests that fis may be involved in 'fine-tuning' the homeostatic control mechanism of DNA supercoiling in E. coli. PMID- 11029699 TI - Knockout of the regulatory site of 3-ketoacyl-ACP synthase III enhances short- and medium-chain acyl-ACP synthesis. AB - 3-ketoacyl-acyl carrier protein synthase (KAS) III catalyses the first condensing step of the fatty acid synthase (FAS) type II reaction in plants and bacteria, using acetyl CoA and malonyl-acyl carrier protein (ACP) as substrates. Enzymatic characterization of recombinant KAS III from Cuphea wrightii embryo shows that this enzyme is strongly inhibited by medium-chain acyl-ACP end products of the FAS reaction, i.e. inhibition by lauroyl-ACP was uncompetitive towards acetyl CoA and non-competitive with regard to malonyl-ACP. This indicated a distinct attachment site for regulatory acyl-ACPs. Based on alignment of primary structures of various KAS IIIs and 3-ketoacyl CoA synthases, we suspected the motif G290NTSAAS296 to be responsible for binding of regulatory acyl-ACPs. Deletion of the tetrapeptide G290NTS293 led to a change of secondary structure and complete loss of KAS III condensing activity. Exchange of asparagine291 to aspartate, alanine294 to serine and alanine295 to proline, however, produced mutant enzymes with slightly reduced condensing activity, yet with insensitivity towards acyl-ACPs. To assess the potential of unregulated KAS III as tool in oil production, we designed in vitro experiments employing FAS preparations from medium-chain fatty acid-producing Cuphea lanceolata seeds and long-chain fatty acid-producing rape seeds, each supplemented with a fivefold excess of the N291D KAS III mutant. High amounts of short-chain acyl-ACPs in the case of C. lanceolata, and of medium-chain acyl-ACPs in the case of rape seed preparations, were obtained. This approach targets regulation and offers new possibilities to derive transgenic or non-transgenic plants for production of seed oils with new qualities. PMID- 11029700 TI - Activation of CDK-activating kinase is dependent on interaction with H-type cyclins in plants. AB - cDNAs encoding cyclin H homologs were isolated from poplar (Populus tremula X tremuloides) and rice (Oryza sativa) plants, and were designated Pt;cycH;1 and Os;cycH;1, respectively. The deduced amino-acid sequences showed 40-60% similarity to human cyclin H and Schizosaccharomyces pombe Mcs2, with higher similarity in the cyclin box region. While Pt;cycH;1 and Os;cycH;1 were expressed in all tissues examined, the transcripts accumulated abundantly in dividing cells. Expression of Os;cycH;1 was abundant in the S-phase in partially synchronized suspension cells, and was induced by submergence in internodes of deepwater rice. A yeast two-hybrid assay demonstrated that both Pt;CycH;1 and Os;CycH;1 were able to interact with rice R2 kinase, which is structurally and functionally similar to cyclin-dependent kinase (CDK)-activating kinase (CAK) of vertebrates. Moreover, an in vitro pull-down assay showed that Os;CycH;1 specifically bound to R2 but not to other rice CDKs. When R2 was expressed in budding yeast CAK mutant, the suppression activity in terms of temperature sensitivity was enhanced by co-expression with Os;cycH;1. Furthermore, in vitro kinase assay indicated that the kinase activities of R2 on CDKs and the carboxy terminal domain (CTD) of the largest subunit of RNA polymerase II were markedly elevated by binding to Os;CycH;1. Our results suggest that cyclin H is a regulatory subunit of CAK, which positively controls CDK- and CTD-kinase activities in plant cells. PMID- 11029701 TI - FRL1 is required for petal and sepal development in Arabidopsis. AB - A novel flower mutant, frl1 (frill 1) was isolated in Arabidopsis thaliana. The frl1 mutant has serrated petals and sepals but the other floral and vegetative organs appear to be normal. To analyse the role of the FRL1 gene, morphological, cytological and double mutant analyses were carried out. The frl1 flower had broader petals and sepals as compared with the wild-type. The distal region of frl1 petals contained fewer epidermal cells but their size was variable and generally larger than that in the wild-type. However, no significant difference was found in the basal region. Observations of the early petal development revealed that the morphology of the developing frl1 petal was normal until the middle of stage 9, but the frl1 phenotype became apparent in stages later than 10. Furthermore, larger nuclei with varied sizes were observed in the distal region of frl1 petals, but not in this region in wild-type petals. This strongly suggests that abnormal endo-reduplication had occurred. These observations indicate that the frl1 mutation affects the number of cell divisions and the subsequent cell expansion during the late stage of petal lamina formation, and that FRL1 might be maintaining the mitotic state or suppressing the transition to the endo-reduplication cycle. Double mutants with the homeotic mutants apetala3-1 and agamous showed additive phenotypes. Ectopic petals in the third whorl of fr11 ag flowers were serrated, indicating that the FRL1 gene acts in petal and sepal development in an organ-specific manner. PMID- 11029702 TI - Antisense-inhibition of ADP-glucose pyrophosphorylase in developing seeds of Vicia narbonensis moderately decreases starch but increases protein content and affects seed maturation. AB - The small subunit of a Vicia faba ADP-glucose pyrophosphorylase (AGP) cDNA was expressed in antisense orientation in Vicia narbonensis under the control of the seed-specific legumin B4 promoter. From several independent transgenic lines both ADP-glucose pyrophosphorylase AGP-mRNA and AGP enzyme activity were reduced by up to 95% in the cotyledons during the mid- to late-maturation phase. Starch was moderately decreased and sucrose was increased. In two of three lines, transcripts encoding the large subunit of AGP and the storage protein vicilin were increased, whereas legumin B-mRNA was decreased. Transcripts of other storage-associated genes were not altered. The cotyledons contained more protein and total nitrogen. Despite the reduction in starch, total carbon was not decreased and dry weight was unchanged. Compared to the wild type, transgenic seeds contained more water and accumulated dry weight during a longer period, and therefore had a prolonged seed-filling period. Transgenic cotyledon cells of comparable age to the wild type were more highly vacuolated and contained smaller starch grains, indicating a delay in cellular differentiation. We conclude that a specific alteration in carbon metabolism can have pleiotropic effects on water and nitrogen content and induces temporal changes in seed development. PMID- 11029703 TI - Characterization and translational regulation of the arginine decarboxylase gene in carnation (Dianthus caryophyllus L.). AB - Arginine decarboxylase (ADC; EC 4.1.1.9) is a key enzyme in polyamine biosynthesis in plants. We characterized a carnation genomic clone, gDcADC8, in which the deduced polypeptide of ADC was 725 amino acids with a molecular mass of 77.7 kDa. The unusually long 5'-UTR that contained a short upstream open reading frame (uORF) of seven amino acids (MQKSLHI) was predicted to form an extensive secondary structure (free energy of approximately -117 kcal mol-1) using the Zuker m-fold algorithm. The result that an ADC antibody detected two bands of 45 and 33 kDa in a petal extract suggested the full length of the 78 kDa polypeptide precursor converted into two polypeptides in the processing reaction. To investigate the role of the transcript leader in translation, in vitro transcription/translation reactions with various constructs of deletion and mutation were performed using wheat germ extract. The ADC transcript leader affected positively downstream translation in both wheatgerm extract and primary transformant overexpressing ADC gene. It was demonstrated that heptapeptide (8.6 kDa) encoded by the ADC uORF was synthesized in vitro. Both uORF peptide, and the synthetic heptapeptide MQKSLHI of the uORF, repressed the translation of downstream ORF. Mutation of the uORF ATG codon alleviated the inhibitory effect. ORF translation was not affected by either a frame-shift mutation in uORF or a random peptide. To our knowledge, this is the first report to provide evidence that a uORF may inhibit the translation of a downstream ORF, not only in cis but also in trans, and that the leader sequence of the ADC gene is important for efficient translation. PMID- 11029704 TI - Transactivation of the Brassica napus napin promoter by ABI3 requires interaction of the conserved B2 and B3 domains of ABI3 with different cis-elements: B2 mediates activation through an ABRE, whereas B3 interacts with an RY/G-box. AB - The transcriptional activator ABI3 is a key regulator of gene expression during embryo maturation in crucifers. In monocots, the related VP1 protein regulates the Em promoter synergistically with abscisic acid (ABA). We identified cis elements in the Brassica napus napin napA promoter mediating regulation by ABI3 and ABA, by analyzing substitution mutation constructs of napA in transgenic tobacco plantlets ectopically expressing ABI3. In transient analysis using particle bombardment of tobacco leaf sections, a tetramer of the distB ABRE (abscisic acid-responsive element) mediated transactivation by ABI3 and ABI3 dependent response to ABA, whereas a tetramer of the composite RY/G complex, containing RY repeats and a G-box, mediated only ABA-independent transactivation by ABI3. Deletion of the conserved B2 and B3 domains of ABI3 abolished transactivation of napA by ABI3. The two domains of ABI3 interact with different cis-elements: B2 is necessary for ABA-independent and ABA-dependent activations through the distB ABRE, whereas B3 interacts with the RY/G complex. Thus B2 mediates the interaction of ABI3 with the protein complex at the ABRE. The regulation of napA by ABI3 differs from Em regulation by VP1, in that the B3 domain of ABI3 is essential for the ABA-dependent regulation of napA. PMID- 11029705 TI - Arabidopsis DNA ligase IV is induced by gamma-irradiation and interacts with an Arabidopsis homologue of the double strand break repair protein XRCC4. AB - Rejoining of single- and double-strand breaks (DSBs) introduced in DNA during replication, recombination, and DNA damage is catalysed by DNA ligase enzymes. Eukaryotes possess multiple DNA ligase enzymes, each having distinct roles in cellular metabolism. Double-strand breaks in DNA, which can occur spontaneously in the cell or be induced experimentally by gamma-irradiation, represent one of the most serious threats to genomic integrity. Non-homologous end joining (NHEJ) rather than homologous recombination is the major pathway for repair of DSBs in organisms with complex genomes, including humans and plants. DNA ligase IV in Saccharomyces cerevisiae and humans catalyses the final step in the NHEJ pathway of DSB repair. In this study we identify an Arabidopsis thaliana homologue (AtLIG4) of human and S. cerevisiae DNA ligase IV which is shown to encode an ATP dependent DNA ligase with a theoretical molecular mass of 138 kDa and 48% similarity in amino-acid sequence to the human DNA ligase IV. Yeast two-hybrid analysis demonstrated a strong interaction between A. thaliana DNA ligase IV and the A. thaliana homologue of the human DNA ligase IV-binding protein XRCC4. This interaction is shown to be mediated via the tandem BRCA C-terminal domains of A. thaliana DNA ligase IV protein. Expression of AtLIG4 is induced by gamma irradiation but not by UVB irradiation, consistent with an in vivo role for the A. thaliana DNA ligase IV in DSB repair. PMID- 11029706 TI - Maize ROP7 GTPase contains a unique, CaaX box-independent plasma membrane targeting signal. AB - Signals in the carboxy-terminal hypervariable region (HVR) of Rho and Ras GTPases target these proteins to specific membrane compartments, where they function in signal transduction. ROP6 and ROP7 are closely related maize Rops (a plant specific Rho subgroup) that share HVR sequences divergent from other Rho HVRs. Both ROPs terminate in CAA, instead of the consensus C-terminal CaaX motif required for membrane association of all characterized Ras and Rho GTPases. The ROP6/7 HVR contains two additional cysteines, potential sites for post translational modification that leads to membrane association; one is in an internal CaaX motif, which would be at the C-terminus if the final intron in both genes were not removed. Transient expression of a GFP-ROP7 fusion revealed its near-total association with the plasma membrane (PM). Furthermore, the ROP7 HVR is sufficient to target GFP to the PM. Surprisingly, the cysteine in the terminal CAA is not required for PM targeting of GFP-ROP7. In contrast, an internal HVR cysteine is essential for proper targeting of the fusion, and the cysteine in the internal CaaX is required for complete membrane association. Interestingly, this CaaX motif can also direct PM association when placed at the fusion C-terminus by addition of an internal stop codon. Fractionation experiments confirm that maize ROPs associate with membranes in maize seedlings. Our analysis suggests that the ROP7 HVR directs PM localization by a mechanism independent of a C-terminal CaaX motif; this mechanism may have evolved through addition of 3' intron/exon sequences to a rop progenitor. PMID- 11029707 TI - Mutations in the WUSCHEL gene of Arabidopsis thaliana result in the development of shoots without juvenile leaves. AB - The vegetative growth of Arabidopsis thaliana can be divided into two phases. The transition from the juvenile (early) phase to the adult (later) phase is associated with changes in several morphological features of leaves, such as the shape of leaf blades, the number of trichomes and patterns of venation. In a screening of mutants with altered morphological identities of leaves, we found one which we named juvenile leafless and misshapen shoot apical meristem (jam). The mutation represented a new allele of the WUSCHEL (WUS) gene, and, in its presence, plants produced no juvenile leaves. Analysis of the morphology of mutant plants revealed that all the rosette leaves had characteristics of adult leaves. The formation of the first rosette leaf in the wus(jam) mutant was markedly delayed, and occurred at the almost same time as formation of the third or fourth leaf in wild-type plants. In the wild-type, these leaves correspond to the first adult leaves. Analysis by RT-PCR showed that transcripts of WUS accumulated in shoot apices and roots, but not in cotyledons and leaves. The present results suggest that the WUS gene controls the morphological traits of rosette leaves either directly or indirectly. In view of the predicted function of the WUS gene, namely maintenance of stem cells within the shoot apical meristem, we suggest that the lack of juvenile leaves in the mutant might have been caused by interruption of leaf initiation during the juvenile phase or by halting of an entire process of formation of juvenile leaves. PMID- 11029708 TI - Cytokinin and gibberellin activate SaMADS A, a gene apparently involved in regulation of the floral transition in Sinapis alba. AB - In plants of Sinapis alba induced to flower by one long day, the MADS box gene, SaMADS A, is expressed initially in the central corpus (L3 cells) of the shoot apical meristem (SAM), about 1.5-2 days before initiation of the first floral meristem. We have combined a physiological approach by testing the effects of three putative floral signals on SaMADS A expression in the SAM of S. alba plants with a transgenic approach using Arabidopsis thaliana plants. A single application of a low dose of a cytokinin or a gibberellin to the apex of vegetative S. alba plants is capable of mimicking perfectly the initial effect of the long day on SaMADS A transcription. A treatment combining the two hormones causes the same activation but seems to enhance the level of SaMADS A expression. A sucrose application to the apex of vegetative plants is, on the contrary, unable to activate SaMADS A expression. None of these chemicals, alone or combined, is capable of causing the floral shift at the SAM. Since the constitutive expression of SaMADS A leads to precocious flowering in A. thaliana and antisense expression of a fragment of the A. thaliana homologue AGL20 leads to a delay in flowering time, these results are consistent with SaMADS A activation being an intermediate event in a cytokinin- and/or gibberellin triggered signal transduction pathway that is involved in the regulation of floral transition in S. alba. PMID- 11029709 TI - Tissue-specific oxylipin signature of tomato flowers: allene oxide cyclase is highly expressed in distinct flower organs and vascular bundles. AB - A crucial step in the biosynthesis of jasmonic acid (JA) is the formation of its correct stereoisomeric precursor, cis(+)12-oxophytodienoic acid (OPDA). This step is catalysed by allene oxide cyclase (AOC), which has been recently cloned from tomato. In stems, young leaves and young flowers, AOC mRNA accumulates to a low level, contrasting with a high accumulation in flower buds, flower stalks and roots. The high levels of AOC mRNA and AOC protein in distinct flower organs correlate with high AOC activity, and with elevated levels of JA, OPDA and JA isoleucine conjugate. These compounds accumulate in flowers to levels of about 20 nmol g-1 fresh weight, which is two orders of magnitude higher than in leaves. In pistils, the level of OPDA is much higher than that of JA, whereas in flower stalks, the level of JA exceeds that of OPDA. In other flower tissues, the ratios among JA, OPDA and JA isoleucine conjugate differ remarkably, suggesting a tissue specific oxylipin signature. Immunocytochemical analysis revealed the specific occurrence of the AOC protein in ovules, the transmission tissue of the style and in vascular bundles of receptacles, flower stalks, stems, petioles and roots. Based on the tissue-specific AOC expression and formation of JA, OPDA and JA amino acid conjugates, a possible role for these compounds in flower development is discussed in terms of their effect on sink-source relationships and plant defence reactions. Furthermore, the AOC expression in vascular bundles might play a role in the systemin-mediated wound response of tomato. PMID- 11029710 TI - Technical advance: identification of plant actin-binding proteins by F-actin affinity chromatography. AB - Proteins that interact with the actin cytoskeleton often modulate the dynamics or organization of the cytoskeleton or use the cytoskeleton to control their localization. In plants, very few actin-binding proteins have been identified and most are thought to modulate cytoskeleton function. To identify actin-binding proteins that are unique to plants, the development of new biochemical procedures will be critical. Affinity columns using actin monomers (globular actin, G-actin) or actin filaments (filamentous actin, F-actin) have been used to identify actin binding proteins from a wide variety of organisms. Monomeric actin from zucchini (Cucurbita pepo L.) hypocotyl tissue was purified to electrophoretic homogeneity and shown to be native and competent for polymerization to actin filaments. G actin, F-actin and bovine serum albumin affinity columns were prepared and used to separate samples enriched in either soluble or membrane-associated actin binding proteins. Extracts of soluble actin-binding proteins yield distinct patterns when eluted from the G-actin and F-actin columns, respectively, leading to the identification of a putative F-actin-binding protein of approximately 40 kDa. When plasma membrane-associated proteins were applied to these columns, two abundant polypeptides eluted selectively from the F-actin column and cross reacted with antiserum against pea annexins. Additionally, a protein that binds auxin transport inhibitors, the naphthylphthalamic acid binding protein, which has been previously suggested to associate with the actin cytoskeleton, was eluted in a single peak from the F-actin column. These experiments provide a new approach that may help to identify novel actin-binding proteins from plants. PMID- 11029711 TI - Atrioventricular plane motion during dobutamine-atropine stress echocardiography: the biphasic response in healthy subjects revisited. AB - This study was undertaken to establish normal values for the systolic atrioventricular plane motion (AVPM) from base to apex during dobutamine-atropine stress echocardiography (DASE) and to compare them with those of patients with coronary artery disease. The AVPM was measured at baseline, low dose and peak dobutamine-atropine infusion in 20 patients referred for coronary angiography and in 20 control subjects. Atrioventricular plane motion was measured at the posterior, anterior, septal, and lateral positions of the mitral annulus in the apical 2- and 4-chamber views by an observer blinded to clinical and angiographic data. In healthy subjects undergoing DASE, AVPM initially increased but subsequently decreased to below baseline values at peak stress. Atrioventricular plane motion at any stage and the changes therein during DASE were within the normal reference interval in the majority of patients. In conclusion, AVPM decreased during DASE in healthy subjects and was not a sensitive marker of coronary artery disease. PMID- 11029712 TI - Differentiation between reversible and irreversible restrictive left ventricular filling patterns with the use of mitral annulus velocity. AB - A restrictive left ventricular filling pattern is generally recognized as an ominous prognostic sign in patients with congestive heart failure. Recently, this filling pattern has been further categorized into reversible and irreversible groups according to the changes in the mitral inflow pattern after preload reduction; furthermore, the prognosis is reported to be different for the two groups. Forty-two patients with a restrictive left ventricular filling pattern who could adequately perform Valsalva's maneuver were studied. Baseline peak early (E) and late (A) mitral inflow velocities, E/A ratio, deceleration time of E velocity, peak early (E') and late (A') diastolic mitral annulus velocities, and E'/A' ratio were obtained. During Valsalva's maneuver, the E/A ratio reversed (<1) in fifteen patients (15/42, 36%). These patients were categorized as belonging to the reversible group. Among the baseline mitral inflow and mitral annulus velocity parameters, A' > 0.05 m/s best discriminated between the reversible and irreversible restrictive left ventricular filling patterns, with a sensitivity of 80% and a specificity of 85%. PMID- 11029713 TI - The relation between mitral annulus motion and ejection fraction: a nonlinear function. AB - In previous studies of the relation between mitral annulus motion (MAM) and left ventricular ejection fraction (EF), a linear relationship has been suggested. In this meta-analysis of 434 patients, we show that the relation is nonlinear and that a linear regression model overestimates EF in the lower range of MAM. The relation is better described by an S function and is influenced by age and heart size. PMID- 11029714 TI - Preload dependence of color M-mode Doppler flow propagation velocity in controls and in patients with left ventricular dysfunction. AB - The purpose of this study was to assess the effects of preload alterations on color M-mode flow propagation velocity (Vp) in volunteers with normal left ventricular (LV) function and in patients with depressed LV function. Color M mode Doppler echocardiography was performed during Valsalva maneuver, passive leg lifting, and after administration of nitroglycerin in 30 healthy volunteers and in 30 age- and sex-matched patients with previous myocardial infarction (MI). Mean Vp in controls was 74 +/- 15 cm/s at baseline and 46 +/- 15 cm/s in MI patients (P <.0005). In both groups, minor changes in Vp were seen during preload alterations; however, these were not significant (control P =.72, MI P =.31). In both groups, peak E-wave velocity (P <.0005), ratio of early-to-late peak velocities (P <.0005), and E-wave deceleration time (P <.0005) were found to change during preload alterations. In conclusion, we found that in controls and patients with previous MI, the color M-mode flow propagation velocity is not affected significantly by preload. PMID- 11029715 TI - Comparison of ventricular volume and mass measurements from B- and C-scan images with the use of real-time 3-dimensional echocardiography: studies in an in vitro model. AB - BACKGROUND: Real-time 3-dimensional (3D) echocardiography avoids geometric assumptions in volume analysis and permits immediate visualization in any plane without the need for cardiac or respiratory gating or computation time. This study compared the accuracy of volume and mass assessments between standard long axis (B-scan) and short-axis (C-scan) views in a simplified but quantifiable left ventricular phantom. METHODS AND RESULTS: The model comprised an inner balloon within an outer balloon separated by ultrasonographic gel. First, to mimic different chamber volumes, 12 volumes (40 to 180 mL) of water within the inner balloon were scanned with a real-time 3D system. Second, 10 volumes (80 to 170 mL) of gel were inserted between the balloons to mimic varying cardiac mass, and the gel volume space (mass) was calculated by subtracting the inner from the outer balloon volume. "Chamber" and "mass" measurements for both B and C scans correlated closely with the actual values (r = 0.99). However, chamber volumes from C scans were consistently less than B-scan values (mean difference from reference for C scans: -5.2 +/- 1.2 mL, P <.0001; for the 2 orthogonal B scans: 0.03 +/- 1.4 mL and -0.9 +/- 1.5 mL, respectively, P = NS). Similarly, for gel volume measurements, B-scan results were closer to actual mass volumes (mean difference 0. 3 +/- 2.5 and 1.7 +/- 2.9 mL) than those of C scans, which tended to underestimate (-4.5 +/- 2.5 mL, P <.0001). CONCLUSION: Our study suggests that real-time 3D echocardiography should provide an accurate means of determining chamber volumes and cardiac mass. However, measurements performed from B-scan views may be closer to the actual values than those from C-scan views, presumably since they are less highly influenced by distortions related to lateral resolution. PMID- 11029716 TI - Superiority of 3-dimensional versus 2-dimensional echocardiography for left ventricular volume assessment in small piglet hearts. AB - To evaluate the accuracy of 3-dimensional (3D) echocardiography in the estimation of left ventricular (LV) volume in vivo, we studied 15 newborn piglets ranging in weight from 2.6 to 11.8 kg. Measurements of beating LV volumes by 3D echocardiograms were compared with measurements by conductance catheter and transthoracic 2-dimensional (2D) echocardiograms with the use of Simpson's rule. The results of both 3D and 2D echocardiograms correlated strongly with the actual volume (r = 0.98 and 0.95 for LV end-diastolic volume, and 0.998 and 0.95 for LV end-systolic volume, respectively). However, the standard error of estimate (SEE) for 2D echocardiography was larger than for 3D. The SEE values for LV end diastolic volume for 2D and 3D echocardiograms were 2.30 mL and 1.85 mL, respectively, and 1.52 mL and 0.5 mL for LV end-systolic volume. We conclude that 3D echocardiography not only accurately measures LV volume and systolic function in a newborn heart, it is more precise than measurements from 2D echocardiography in the assessment of small beating hearts. PMID- 11029717 TI - Three-dimensional imaging of aortic arch anomalies in infants and children with intravascular ultrasound catheters from a transesophageal approach. AB - Aortic arch anomalies usually require surgical intervention preceded by precise anatomic definition. We studied 20 patients to evaluate the feasibility and accuracy of using intravascular ultrasound catheters from a transesophageal approach with 3-dimensional image reconstruction for the diagnosis of aortic arch anomalies in infants and children. All patients had transthoracic echocardiograms and/or angiograms or magnetic resonance imaging. A 12.5-MHz intravascular ultrasound catheter was positioned in the esophagus and withdrawn by using an electrocardiogram and a respiratory gated pullback device to acquire the mediastinal images. All patients with arch anomalies underwent surgical repair. Reconstructed images were analyzed in the "anyplane" mode and with surface rendering. Intravascular ultrasound 3-dimensional imaging was successfully accomplished without complications. Anatomy was correctly identified in all patients by both blinded and unblinded observers, thus confirming the sensitivity and accuracy of the technique. We foresee this new technique to be useful as an adjunctive imaging modality applicable at the bedside or in the cardiac imaging laboratory. PMID- 11029718 TI - Evaluation of regional wall motion and quantitative measures of ventricular function during dobutamine stress echocardiography in pediatric cardiac transplantation patients. AB - BACKGROUND: Graft coronary disease is a leading cause of death in patients who have undergone cardiac transplantation. The purpose of this study was to evaluate regional wall motion response and quantitative measures of ventricular function during dobutamine stress echocardiography (DSE) in pediatric transplantation patients. METHODS: Eleven patients were evaluated the first year after transplantation (10/11 no rejection). Ten of the 11 were reevaluated 1.2 +/- 0.3 years later (9/10 no rejection). RESULTS: Dobutamine stress echocardiography revealed the following: (1) baseline regional wall motion abnormalities in 80% that resolved in all, (2) increased heart rate and blood pressure, (3) no change in left ventricular end-diastolic diameter, (4) decreased end-systolic diameter, (5) decreased wall stress and increased velocity of circumferential fiber shortening, (6) increased VCFcZ score (representing systolic left ventricular function), and (7) a decreased mitral passive-to-active filling ratio. Patients with rejection had abnormal VCFcZ scores at rest. CONCLUSIONS: Pediatric transplantation patients without rejection have baseline regional wall motion abnormalities. With DSE, the following are present: (1) resolution of wall motion abnormalities, (2) increased contractility independent of load, and (3) changes in diastolic parameters that reflect increased heart rate. Patients with rejection may have abnormal contractility at rest. PMID- 11029719 TI - Echocardiographic assessment of stentless aortic bioprosthetic valves. AB - Stentless tissue aortic valves are gaining in popularity because of advantages in hemodynamics and durability compared with stented bioprostheses. The absence of a rigid sewing ring and struts makes these valves pliable, and distortion at implantation can result in valve dysfunction. Because the anatomy and implantation techniques of stentless tissue valves are unlike those of mechanical and stented tissue valves, their echocardiographic appearance is unique on both intraoperative and subsequent transthoracic and transesophageal echocardiography. This report describes the echocardiographic appearance of normally functioning stentless tissue heterograft aortic valves as an aid to their intraoperative and subsequent echocardiographic assessment. PMID- 11029720 TI - Migration of a transjugular intrahepatic portosystemic shunt (TIPS) stent: evaluation by transesophageal echocardiography. AB - Transjugular intrahepatic portosystemic shunting (TIPS) is a procedure for end stage liver disease that involves angiographically guided placement of an intrahepatic expandable metal stent. Mechanical complications of intrahepatic stent placement have been reported, including stent migration to the central venous circulation. This report describes a patient who had embolization of a stent after a TIPS procedure, with subsequent failed percutaneous attempts at stent removal. Transesophageal echocardiography documented the stent caught in the tricuspid valve and apparatus, with its distal end projecting into the right ventricular cavity. PMID- 11029721 TI - Fatal cardiac rupture: a case of subepicardial aneurysm after myocardial infarction. AB - We report a case in which 2-dimensional echocardiography established the diagnosis of a left ventricular subepicardial aneurysm that was followed by rupture and sudden death before surgery. Two-dimensional echocardiography is of great help in detecting this rare complication after myocardial infarction. Urgent surgical treatment is warranted for this condition. PMID- 11029722 TI - Unsuspected embolic fungal endocarditis of an aortic conduit diagnosed by transesophageal echocardiography. AB - A 66-year-old man with a history of hypertension and ascending aortic replacement because of a type A dissection had 3 successive embolic events (left lower limb, brain, and spleen). Two consecutive transesophageal echocardiography studies showed mobile masses in the ascending aorta. The patient was reoperated without a certain etiologic diagnosis, and an unsuspected fungal endocarditis caused by an unusual germ (Trichoderma species) was found. Transesophageal echocardiography proved very useful in the management of this uncommon case of endocarditis. PMID- 11029723 TI - Echocardiographic demonstration of rupture of intraseptal hydatid cyst. AB - We report a case of an intracardiac hydatid cyst observed before and after rupture complicated by a pulmonary embolus. The echocardiographic findings are analyzed and discussed. This dramatic course underscores the surgical emergency of cardiac hydatidosis. PMID- 11029724 TI - Ultrasound contrast physics: A series on contrast echocardiography, article 3. PMID- 11029725 TI - Manuscript reviewers: the backbone of a peer-reviewed journal PMID- 11029727 TI - Direction of growth PMID- 11029726 TI - Direction of growth. PMID- 11029728 TI - What patients want, and what they need. PMID- 11029729 TI - Longitudinal assessment of vertical and sagittal control in the mandibular arch by the mandibular fixed lingual arch. AB - The purpose of this investigation was to determine and quantify vertical changes in the position of the mandibular molars while maintaining arch perimeter with a fixed lingual arch. Twenty-three patients with a mean age of 10.4 +/- 0.6 years were selected to receive fixed lingual arch treatment as the only appliance in the mandibular arch. Average treatment time was 18.3 +/-+/- 0.6 months. Longitudinal records for 12 and 24 months of 24 individuals matched by ethnic origin, age, gender, and mandibular plane inclination were used as controls. Pretreatment and posttreatment cephalograms were used to determine positional changes. Statistically significant differences between the fixed lingual arch and control groups were found. The results of this investigation indicated that the mandibular fixed lingual arch is a useful tool to control the vertical development of the mandibular molars. PMID- 11029730 TI - Ratings of profile attractiveness after functional appliance treatment. AB - The aim of this study was to determine the change in profile attractiveness in children with Class II Division 1 malocclusion after 18 months' treatment with functional appliances. Changes in profile attractiveness were assessed by panels of art students, dental students, and parents of orthodontic patients. Each panel consisted of an equal number of male and female raters. The raters first decided whether the initial or 18-month profile silhouette was more attractive, and then scored the degree to which it was more attractive on an unmarked visual analog scale. There were no significant differences between either male and female raters or among panels in their assessments of the change in profile attractiveness in the whole sample. Neither were there significant differences between the change in profile attractiveness of the untreated subjects and the subjects treated with either Frankel function regulators or Harvold activators. It is concluded that treatment with functional appliances does not lead to more attractive profiles than nontreatment. PMID- 11029731 TI - On "Ratings of facial attractiveness after functional appliance treatment" PMID- 11029732 TI - Cephalometric appraisal of posttreatment vertical changes in adult orthodontic patients. AB - The purpose of this study was to evaluate vertical facial changes in adult orthodontic patients and to evaluate the stability of these changes. Thirty-three patients (8 males and 25 females) were examined. The patients had been treated with full fixed edgewise appliance mechanics and exhibited at least 1.0 degrees of clockwise rotation of the mandible during treatment. Mandibular rotation was determined by the angular change in the Y-axis to the Frankfort plane. Twelve angular and 14 linear skeletal and dental measurements and 3 skeletal ratios were derived from pretreatment (T1), posttreatment (T2), and postretention (T3) cephalometric radiographs. Paired t tests were used to compare vertical changes that occurred as a result of orthodontic treatment and their stability or relapse tendency during the retention and postretention periods. Twenty-five percent (P <.001) of the opening rotation of the mandible recovered during the posttreatment period, resulting in a significant overall rotation that was maintained. Both treatment and posttreatment changes in the Y-axis angle showed a high correlation with the horizontal position of pogonion (r = -0.797 and -0.889, respectively). Only overjet showed a low correlation between treatment changes and posttreatment changes in the Y-axis angle. Stepwise regression analysis of pretreatment variables and treatment changes failed to predict the behavior of the Y-axis angle change. PMID- 11029733 TI - Oral perception in tongue thrust and other oral habits. AB - Oral stereognosis is the ability of the mouth to recognize shape and texture. Oral shape recognition is sensitive to repetition and to topical mucosal anesthesia. Age, upper and lower arch perimeter, and labiolingual dysfunction also interact with oral stereognosis. The purpose of this investigation was to define the influence of age, arch size, and oral dysfunction on oral stereognosis when submitted to repeated trials. Fifty subjects were selected before orthodontic treatment. Each subject underwent 4 trials: T1 and T4 without anesthesia and strictly similar, T2 with topical anesthesia of the tongue, and T3 with topical anesthesia of the palate. Five test pieces or stimuli were used. The recognition time (RT) of each stimulus, the perimeter of upper and lower anterior dental arch, and the labiolingual dysfunction index (LLDI) were the main variables statistically evaluated. Subjects with a mild degree of dysfunction needed more time to recognize the stimuli in T3 when compared with T2. The number of RT3 > RT2 was 2.5 +/- 1.12 in the group with a low LLDI (12 +/- 1.5), and 1.57 +/- 0.63 in the group with an LLDI of 16 +/- 2.5 (P >.05). This may be attributed to different manipulation of the test pieces between the 2 groups, which could have been modified through sensory deprivation. Bolus recognition before the swallowing act needs to be paralleled to stereognostic performance. PMID- 11029734 TI - Effects of twin-block therapy on protrusive muscle functions. AB - Protrusive mandibular function, including maximum protrusive force and fatigue time, was investigated in 66 children displaying Class II Division 1 malocclusion. Thirty-two children were treated with the Clark Twin-block appliance and the other 34 children served as untreated controls. The observation period was 6 months. Cross-sectional data based on pretreatment records showed that maximum protrusive force ranged from 18.5 N to 160 N, with a mean of 80.3 +/ 30.7 N. Maximum protrusive force was significantly higher in males than in females (P <.001). The correlation between maximum protrusive force and chronologic age was low (r = 0.20) and did not reach significance. Maximum protrusive force in the group of children with disk displacement was not significantly different from that of the group without disk displacement. Comparison of pretreatment and 6-month records in the untreated control group revealed a significant increase in maximum protrusive force (P <.01) as a result of normal growth, while the measured change in the Twin-block-treated children did not reach significance. Fatiguing the protrusive muscles did not alter mandibular position in the Twin-block group after 6 months of treatment. The present study does not support the lateral pterygoid hypothesis, as there was no evidence of an increase in mandibular protrusive function after treatment with the Twin-block functional appliance. PMID- 11029735 TI - Stability of bilateral sagittal split ramus osteotomy: rigid fixation versus transosseous wiring. AB - Although many improvements have been made in orthodontic surgical procedures for mandibular retrognathism, relapse continues to occur. This study was designed to compare the stability of rigid and nonrigid fixation between 2 groups of patients who had undergone mandibular advancement surgery via sagittal split ramus osteotomy. Retrospective cephalometric measurements were made on 54 randomly selected orthognathic surgical patients. The patients, 7 males and 47 females, were divided into 2 groups: 28 patients stabilized by means of rigid fixation and 26 patients fixated with interosseous wires. The age of the patients ranged from 15.3 to 49.7 years. Lateral cephalograms were used to evaluate each patient at 3 distinct intervals: 7.0 +/- 2.0 days before surgery (T1), 34.4 +/- 15.0 days postsurgery (T2), and 458 +/- 202 days after sagittal split osteotomy (T3). Eighteen linear and angular measurements were recorded and differences between the 3 time periods were evaluated. Statistical analyses were performed to assess the differences in the 2 fixation types between and within each group at different time intervals. The following measurements showed statistically significant skeletal relapse over time, for the P value.0028: Co-Go, ANS-Xi-Pm, IMPA, overbite, and overjet. The remaining variables showed no statistically significant relapse. The only measurement that showed a statistically significant group difference between T1 and T2 was DC-Xi-Pm. Results of the study led to the following conclusions: there was statistically significant relapse in mandibular length, lower anterior face height, mandibular arc, lower incisor inclination, overbite, and overjet in each group, regardless of the type of fixation. The potential was greater for relapse in patients stabilized with transosseous wiring. Although multifactorial, relapse in overbite and overjet may be a combination of skeletal and dental changes. (Am J Orthod Dentofacial Orthop 2000;118:397-403). PMID- 11029736 TI - Treatment and posttreatment craniofacial changes after rapid maxillary expansion and facemask therapy. AB - The aim of this study was to evaluate treatment and posttreatment dentoskeletal changes in 2 groups of subjects with Class III malocclusions. Subjects were treated with a bonded acrylic-splint expander and a face mask, and the optimal timing for this treatment protocol was assessed. The treated sample (29 subjects) was divided into 2 groups according to the stage of dental development. The early treatment group consisted of 16 subjects in the early mixed dentitional (erupting permanent incisors and/or first molars), whereas the late treatment group consisted of 13 subjects in the late mixed dentition (erupting permanent canines and premolars). Cephalograms were available at 3 time periods: T(1), pretreatment, T(2), end of active treatment, and T(3), posttreatment. The mean T(1)-T(2) interval (active treatment period) and the mean T(2)-T(3) interval (posttreatment period) were approximately 1 year each in both treatment groups. None of the patients wore any skeletal retention appliance during the posttreatment period (T(2)-T(3)). Groups of subjects with untreated Class III malocclusion were used as controls at both observation intervals. A significant increase in the sagittal growth of the maxilla was seen only when treatment was performed in the early mixed dentition. A restraining effect on mandibular growth rate associated with a more upward and forward direction of condylar growth was found in both treatment groups. An increase in vertical intermaxillary relationships was observed in Class III patients treated in the late mixed dentition. Posttreatment, the Class III craniofacial growth pattern was re established in the absence of any skeletal retention appliance. Relapse tendency affects the sagittal growth of the maxilla in the early treated subjects and the sagittal position of the mandible in the late treated subjects. Orthopedic treatment of Class III malocclusion in the early mixed dentition is able to induce more favorable craniofacial adaptations than treatment in the late mixed dentition. PMID- 11029737 TI - Evaluation of treatment and posttreatment changes of protraction facemask treatment using the PAR index. AB - The purpose of this study was to use the Peer Assesment Rating (PAR) index score to evaluate the treatment and posttreatment changes of Class III patients treated by protraction facemask. The sample consisted of 20 Chinese children, 6 to 11 years old, with Class III skeletal malocclusion who had been treated with maxillary expansion and a protraction facemask. The average treatment time was 8.2 months, followed by 1 year of retention with a Class III functional appliance. Study casts were taken pretreatment (T1), posttreatment (T2), 1 year follow-up (T3), and 2 years follow-up (T4). After treatment, PAR scores were calculated for each time period. Differences among the 4 time periods were analyzed with the Wilcoxin matched-pairs test. Significant reductions in PAR scores were found at T2 (56%), T3 (70%), and T4 (63%) compared with T1. Immediately posttreatment (T2), 17 (85%) of 20 patients had improved PAR scores by a reduction of at least 30%. Reductions were caused primarily by correction of the anterior crossbite. One year after treatment (T3), further reductions in PAR score were noted (P <.01) as a result of better alignment of the anterior segment, improvement of the buccal occlusion, and overbite and midline corrections. Two years after treatment (T4), PAR scores were higher than at the previous time period. The increases were due to relapses in overjet (4 of 20 patients), overbite, and centerline corrections. These results indicate that significant reductions in the severity of Class III malocclusion can be achieved with early orthopedic facemask treatment. In most cases, further improvement in the PAR score can be expected 1 and 2 years after treatment. In a few patients, the benefits of early treatment are negated by relapses in overjet, overbite, and centerline corrections during the follow-up period. PMID- 11029738 TI - The relationship of 2 professional occlusal indexes with patients' perceptions of aesthetics, function, speech, and orthodontic treatment need. AB - The aim of this study was to determine the relationship between patients' perceptions of orthodontic treatment need and need as determined with professionally derived indexes, namely the dental aesthetic index and the index of orthodontic treatment need. This study was undertaken at orthodontic offices in San Francisco, Calif. The pretreatment study casts of 50 consecutive patients, presenting for orthodontic treatment, were objectively assessed with these indexes by 2 examiners trained and calibrated in their use. Patients were asked to complete a questionnaire consisting of 4 questions addressing appearance, function, speech, and treatment need, using either a 5-point Likert scale or a yes/no response. The professionally derived indexes showed that statistically significant correlations existed between the aesthetic component and dental health component (r = 0.46; P <.01), the aesthetic component and dental aesthetic index (r = 0.54; P <.01), and the dental health component and dental aesthetic index (r = 0.46; P <.01). Statistically significant correlations were also found for subjective assessments between biting/chewing and speech (r = 0.31; P <.05), between speech and the aesthetic component (r = -0.39; P <. 01) and the dental aesthetic index (r = 0.34; P <.05), and between the aesthetic component and appearance (r = -0.28; P <.05). Logistic regression analysis after dichotomization (treatment/no treatment need) confirmed that the aesthetic component was the only statistically significant factor (odds ratio, 0.57; 95% confidence limits 0.34 to 0.97). PMID- 11029739 TI - Location of the apex of the lower central incisor. AB - Clinicians have always had difficulty finding the apex of the lower central incisor on a lateral cephalometric radiograph. This study was undertaken to define the radiographic anatomy surrounding the true apex. Standard cephalograms were taken of 38 skulls with metallic markers in the socket apex of 1 lower central incisor. Similar cephalograms were taken with the lower central incisor apex positioned in the central ray of the x-ray source. The comparison of the marked apices in the complete skulls and their separate mandibles showed that there was no distortion. Measurements from the markers to the lingual and labial symphyseal borders located the apices labial of center. PMID- 11029740 TI - Radiographic localization of unerupted mandibular anterior teeth. AB - The parallax method and the use of 2 radiographs taken at right angles to each other are the 2 methods generally used to accurately localize teeth. For the parallax method, the combination of a rotational panoramic radiograph with an occlusal radiograph is recommended. This combination involves a vertical x-ray tube shift. Three case reports are presented that illustrate: (1) how this combination can accurately localize unerupted mandibular anterior teeth, (2) how a deceptive appearance of the labiolingual position of the unerupted tooth can be produced in an occlusal radiograph, (3) how increasing the vertical angle of the tube for the occlusal radiograph makes the tube shift easier to discern, (4) why occlusal radiographs are preferable to periapical radiographs for tube shifts, and (5) how localization can also be carried out with 2 radiographs at right angles to each other, one of which is an occlusal radiograph taken with the x-ray tube directed along the long axis of the reference tooth. PMID- 11029741 TI - Radiographic localization of unerupted teeth: further findings about the vertical tube shift method and other localization techniques. AB - The parallax method (image/tube shift method, Clark's rule, Richards' buccal object rule) is recommended to localize unerupted teeth. Richards' contribution to the development of the parallax method is discussed. The favored method for localization uses a rotational panoramic radiograph in combination with an occlusal radiograph involving a vertical shift of the x-ray tube. The use of this combination when localizing teeth and supernumeraries in the premolar region is illustrated. When taking an occlusal radiograph to localize an unerupted maxillary canine, clinical situations are presented where modification of the vertical angulation of the tube of 70 degrees to 75 degrees or of the horizontal position of the tube is warranted. The limitations of axial (true, cross sectional, vertex) occlusal radiographs are also explored. PMID- 11029742 TI - The Royal London Space Planning: an integration of space analysis and treatment planning: Part I: Assessing the space required to meet treatment objectives. AB - The Royal London Space Planning process has evolved since 1985 to ensure a disciplined approach to diagnosis and treatment planning and to provide a record to justify treatment decisions for professional accountability. The analysis takes into consideration most aspects of a given malocclusion and aims to quantify the space required in each dental arch to attain the treatment objectives. Space planning also helps determine whether the objectives are likely to be attainable and helps in the planning of treatment mechanics and the control of anchorage. The process of analysis is divided into 2 sections. The first part consists of assessing the original malocclusion according to various component parts, any of which may have an effect on space if altered during treatment. These components are crowding and spacing, occlusal curves, arch width, anteroposterior position of labial segments, mesiodistal angulation, and incisor inclination. The second part of the analysis, which will be published in a separate article, deals with the effect of treatment procedures, such as extractions, tooth-size modifications, distal or mesial molar movements, as well as natural growth, on the space required. Space planning should be regarded only as a useful guide, as many areas of orthodontics-including growth, biological response, and patient compliance-cannot be controlled with total accuracy. PMID- 11029743 TI - The Royal London Space Planning: an integration of space analysis and treatment planning: Part II: The effect of other treatment procedures on space. AB - The Royal London Space Planning process is carried out in 2 stages. The first stage, assessing the space required to attain the treatment objectives, was described in Part I of this report, published earlier. In Part II, the process of integrating space analysis with treatment planning continues with consideration of the effects other treatment procedures have on space. These procedures include tooth enlargement or reduction, tooth extraction, the creation of space for prosthetic replacement, and mesial and distal molar movement. The effects of favorable and unfavorable growth are also considered. A brief case report is presented to demonstrate use of the Royal London Space Planning. PMID- 11029745 TI - Earn 3 hours of CE credit PMID- 11029744 TI - Asymmetric extraction treatment of an Angle Class II Division 2 subdivision left malocclusion with anterior and posterior crossbites. AB - This case was chosen by the CDABO student case selection committee for publication in the AJO/DO. PMID- 11029746 TI - Digital image processing: how to retouch your clinical photographs. PMID- 11029747 TI - Litigation, legislation, and ethics. Informed consent and the fourth dimension. PMID- 11029748 TI - Angiogenesis in the thyroid gland. AB - Angiogenesis is the mechanism of blood vessel formation after the first few days of embryogenesis, and is essential for all tissue growth. In adults, angiogenesis occurs in the thyroid during disease processes including goitre, Graves' disease, thyroiditis and cancer. The molecular mechanisms controlling angiogenesis are becoming clearer, and therapy targeting these processes is coming closer to clinical fruition. Both promoters and inhibitors of angiogenesis have been identified in the thyroid, including vascular endothelial growth factor (VEGF), fibroblast growth factor, and thrombospondin. This commentary will review the understanding of the control of angiogenesis within the context of the thyroid gland, and review the pre-eminent role of VEGF as the angiogenic signal from the follicular cells to the endothelial cells. PMID- 11029749 TI - Carnitine: an osmolyte that plays a metabolic role. AB - Carnitine, gamma-trimethyl-beta-hydroxybutyrobetaine, is a small molecule widely present in all cells from prokaryotic to eukaryotic ones. It is the sole source of carbon and nitrogen in some bacteria; it serves as osmoprotectant in others. It is a carrier of acyl moieties, and exclusively of long-chain fatty acids for mitochondrial beta-oxidation in mammals. The conspicuously similar composition of the intracellular milieu among widely different species in relation to organic osmolyte systems involves the methylamine family to which carnitine belongs. This prompted us to examine the osmolytic properties of carnitine in an attempt to clarify the metabolic functions carnitine has acquired during evolution. An understanding of the metabolic functions of this organic compatible solute impinge on research involving this compound. PMID- 11029750 TI - Modulation of the ERK pathway of signal transduction by cysteine proteinase inhibitors. AB - Cell proliferation requires the coordinate synthesis and degradation of many proteins. In addition to the well-characterized involvement of the proteasome in the degradation of several cell cycle-regulated proteins, it has been established that cysteine proteinases are also involved in the control of cell proliferation, but their role is currently not understood. By using both synthetic cysteine proteinase inhibitors and overexpression of T-kininogen (T-KG), a physiologically relevant cysteine proteinase inhibitor, we show that inhibition of cysteine proteinases results in a severe inhibition of the ERK pathway of signal transduction. Mechanistically, this effect appears to be the result of stabilization of the ERK phosphatase MKP-1, which leads to an enhanced dephosphorylation (and hence inactivation) of ERK molecules. These results are specific to cysteine proteinase inhibitors and are not observed when either serine proteinases or the proteasome are inhibited. We hypothesize that inhibition of cysteine proteinases in vivo leads to a dysregulation of the ERK pathway, which results in an inability of the cell to transmit to the nucleus the signals generated by the presence of growth factors, thus resulting in loss of cell proliferation. PMID- 11029751 TI - Differential gene expression of rat neonatal heart analyzed by suppression subtractive hybridization and expressed sequence tag sequencing. AB - Heart diseases have been one of the major killers among the human population worldwide. Because the vast majority of cardiomyocytes cannot regenerate once they cease to proliferate shortly after birth, functionally significant myocardial regeneration is not observed clinically. Whether these cells are terminally differentiated and permanently withdrawn from the cell cycle is controversial, but broadening our understanding of the rapid switch from hyperplastic to hypertrophic growth of cardiomyocytes during neonatal myocardial development may shed light on novel cardiovascular therapies. By suppression subtractive hybridization (SSH) and expressed sequence tag (EST) sequencing, we analyzed the differential gene expression of rat neonatal heart. SSH yielded subtracted and normalized cDNA libraries and enhanced the probability of detecting ESTs, which represent genes pertinent to signal transduction/cell regulation and replication/transcription/translation machinery, as compared to the traditional EST sequencing of heart cDNA libraries. PMID- 11029752 TI - GTP-dependent permeabilized neutrophil secretion requires a freely diffusible cytosolic protein. AB - Guanosine triphosphate (GTP) has been implicated in the regulation of Ca(2+) mediated secretion from neutrophils. We further examined the role of GTP in neutrophil secretion using streptolysin O permeabilized cells. We found that, in the presence of GTP, 1.0 microM free Ca(2+) causes maximum secretion-equivalent to that achieved with 100 microM free Ca(2+)-whereas GTPgammaS inhibits Ca(2+) stimulated secretion. Interestingly, GTP by itself stimulates secretion. These results indicate the existence of a GTP-regulated mechanism of secretion in neutrophils that requires GTP hydrolysis to stimulate secretion in the presence and absence of Ca(2+). The stimulatory effect of GTP is only observed when GTP is present during permeabilization. Addition of GTP after permeabilization, when the cytosolic contents have leaked out from cells, gives no stimulatory response, implying that the GTP-dependent secretory apparatus requires at least one cytosolic protein. GTP-dependent secretion can be reconstituted with crude HL-60 and bovine liver cytosol. The reconstituting activity binds to GTP-agarose, suggesting that the cytosolic factor is a GTP-binding protein or forms a complex with a GTP-binding protein. However, it is not a member of the rho or rac families of GTPases. By gel filtration chromatography, the secretion reconstituting activity eluted at 870 and 200 kDa, but in the presence of GTP, eluted at 120 kDa, indicating that it is part of a high-molecular-weight complex that dissociates in the presence of GTP. Retention of adenosine diphosphate ribosylation factor (ARF) in permeabilized cells and insensitivity of the cytosolic reconstituting activity to brefeldin A led to our speculation that ARF6 may be the GTPase involved in GTP-dependent secretion, and that activity from a BFA-insensitive ARF6 guanine nucleotide exchange factor reconstitutes secretion. PMID- 11029753 TI - Enhancement by sphingosine 1-phosphate in vasopressin-induced phosphoinositide hydrolysis in aortic smooth-muscle cells: involvement of p38 MAP kinase. AB - We previously reported that sphingosine 1-phosphate (S-1-P), a sphingomyelin metabolite, activates p44/p42 mitogen-activated protein (MAP) kinase and p38 MAP kinase in aortic smooth-muscle A10 cells. In the present study, we investigated the effect of sphingomyelin metabolites on phospholipase C-catalyzing phosphoinositide hydrolysis induced by arginine vasopressin (AVP) in A10 cells. C(2)-ceramide and sphingosine had little effect on inositol phosphate (IP) formation stimulated by AVP. S-1-P, which alone slightly stimulated the IPs formation, dose-dependently amplified the AVP-induced formation of IPs. Tumor necrosis factor-alpha enhanced the AVP-induced formation of IPs. However, S-1-P did not enhance the formation of IPs by NaF, a heterotrimeric GTP-binding protein activator. Pertussis toxin inhibited the effect of S-1-P. PD98059, an inhibitor of the upstream kinase that activates p44/p42 MAP kinase, had little effect on the enhancement by S-1-P. SB203580, an inhibitor of p38 MAP kinase, suppressed the effect of S-1-P on the formation of IPs by AVP. SB203580 inhibited the AVP induced phosphorylation of p38 MAP kinase. Pertussis toxin suppressed the phosphorylation of p38 MAP kinase by S-1-P. These results indicate that S-1-P amplifies AVP-induced phosphoinositide hydrolysis by phospholipase C through p38 MAP kinase in vascular smooth-muscle cells. PMID- 11029754 TI - N-terminal cleavage of bax by calpain generates a potent proapoptotic 18-kDa fragment that promotes bcl-2-independent cytochrome C release and apoptotic cell death. AB - Upon apoptosis induction, the proapoptotic protein Bax is translocated from the cytosol to mitochondria, where it promotes release of cytochrome c, a caspase activating protein. However, the molecular mechanisms by which Bax triggers cytochrome c release are unknown. Here we report that before the initiation of apoptotic execution by etoposide or staurosporin, an active calpain activity cleaves Bax at its N-terminus, generating a potent proapoptotic 18-kDa fragment (Bax/p18). Both the calpain-mediated Bax cleavage activity and the Bax/p18 fragment were found in the mitochondrial membrane-enriched fraction. Cleavage of Bax was followed by release of mitochondrial cytochrome c, activation of caspase 3, cleavage of poly(ADP-ribose) polymerase, and fragmentation of DNA. Unlike the full-length Bax, Bax/p18 did not interact with the antiapoptotic Bcl-2 protein in the mitochondrial fraction of drug-treated cells. Pretreatment with a specific calpain inhibitor calpeptin inhibited etoposide-induced calpain activation, Bax cleavage, cytochrome c release, and caspase-3 activation. In contrast, transfection of a cloned Bax/p18 cDNA into multiple human cancer cell lines targeted Bax/p18 to mitochondria, which was accompanied by release of cytochrome c and induction of caspase-3-mediated apoptosis that was not blocked by overexpression of Bcl-2 protein. Therefore, Bax/p18 has a cytochrome c-releasing activity that promotes cell death independent of Bcl-2. Finally, Bcl-2 overexpression inhibited etoposide-induced calpain activation, Bax cleavage, cytochrome c release, and apoptosis. Our results suggest that the mitochondrial calpain plays an essential role in apoptotic commitment by cleaving Bax and generating the Bax/p18 fragment, which in turn mediates cytochrome c release and initiates the apoptotic execution. PMID- 11029755 TI - Initiation of DNA replication in Saccharomyces cerevisiae G1-phase nuclei by Xenopus egg extract. AB - Xenopus egg extracts initiate replication at specific origin sites within mammalian G1-phase nuclei. Similarly, S-phase extracts from Saccharomyces cerevisiae initiate DNA replication within yeast nuclei at specific yeast origin sequences. Here we show that Xenopus egg extracts can initiate DNA replication within G1-phase yeast nuclei but do not recognize yeast origin sequences. When G1 phase yeast nuclei were introduced into Xenopus egg extract, semiconservative, aphidicolin-sensitive DNA synthesis was induced after a brief lag period and was restricted to a single round of replication. The specificity of initiation within the yeast 2 microm plasmid as well as in the vicinity of the chromosomal origin ARS1 was evaluated by neutral two-dimensional gel electrophoresis of replication intermediates. At both locations, replication was found to initiate outside of the ARS element. Manipulation of both cis- and trans-acting elements in the yeast genome before introduction of nuclei into Xenopus egg extract may provide a system with which to elucidate the requirements for vertebrate origin recognition. PMID- 11029756 TI - Overexpressed poly(ADP-ribose) polymerase delays the release of rat cells from p53-mediated G(1) checkpoint. AB - We have previously reported that in cells ectopically expressing temperature sensitive p53(135val) mutant, p53 formed tight complexes with poly(ADP-ribose) polymerase (PARP). At elevated temperatures, p53(135val) protein, adopting the mutant phenotype, was localized in the cytoplasm and sequestered the endogenous PARP. To prove whether an excess of p53(135val) protein led to this unusual intracellular distribution of PARP, we have established cell lines overexpressing p53(135val) + c-Ha-ras alone or in combination with PARP. Interestingly, immunostaining revealed that PARP is sequestered in the cytoplasm by mutant p53 in cells overexpressing both proteins. Simultaneous overexpression of PARP had no effect on temperature-dependent cell proliferation and only negligibly affected the kinetics of p53-mediated G(1) arrest. However, if the cells were completely growth arrested at 32 degrees C and then shifted up to 37 degrees C, coexpressed PARP dramatically delayed the reentry of transformed cells into the cell cycle. Even after 72 h at 37 degrees C the proportion of S-phase cells was reduced to 20% compared to those expressing only p53(135val) + c-Ha-ras. The coexpressed PARP stabilized wt p53 protein and its enzymatic activity was necessary for stabilization. PMID- 11029757 TI - Antiangiogenesis efficacy of nitric oxide donors. AB - Angiogenesis is a complex process involving endothelial cell migration, proliferation, invasion, and tube formation. Inhibition of these processes might have implications in various angiogenesis-mediated disorders. Because nitric oxide (NO) is known to play a key role in various vascular diseases, the present study was undertaken to determine the role of NO in angiogenesis-mediated processes using the NO donor, S-nitroso N-acetyl penicillamine (SNAP) and S nitroso N-acetyl glutathione (SNAG). The antiangiogenic efficacy of these NO donors was examined using in vivo and in vitro model systems. The in vitro studies demonstrated the ability of SNAP to inhibit cytokine fibroblast growth factor (FGF2)-stimulated tube formation and serum-induced cell proliferation. The inhibitory effect on cell proliferation by SNAP concentrations above the millimolar range was associated with significant shifts in the concentration of NO metabolites. Furthermore, using the mouse Matrigel implant model and the chick chorioallantoic membrane (CAM) models, SNAP demonstrated maximal inhibitory efficacy (85-95% inhibition) of cytokine (FGF2)-induced neovascularization in both in vivo models. SNAP and SNAG resulted in 85% inhibition of FGF2-induced neovascularization in the mouse Matrigel model when given at 5 mg/kg/day infusion in minipumps during 14 days and 87% inhibition of angiogenesis induced by FGF2 in the CAM when administered a single dose of 50 microg. Thus, NO donors might be a useful tool for the inhibition of angiogenesis associated with human tumor growth, or neovascular, ocular, and inflammatory diseases. PMID- 11029758 TI - Increased lymphocytic aminopeptidase N/CD13 promoter activity after cell-cell contact. AB - Aminopeptidase N (APN)/CD13 is a transmembrane ectoenzyme expressed on a wide variety of cells. With respect to haematopoietic cells, APN/CD13 has been considered specific for the myeloid lineage, because granulocytes and monocytes/macrophages, but not lymphocytes of peripheral blood, show a surface expression of CD13 antigen. However, we could recently show that cell-cell contact of lymphocytes with endothelial cells, monocytes, and fibroblast-like synoviocytes (SFCs) results in an increase of steady-state APN/CD13 mRNA and a rapid expression of cell-surface protein on the lymphocytes. In this study using the Dual-Luciferase reporter assay, we demonstrate that interaction of the T-cell line Jurkat with SFCs results in a higher activity of the APN/CD13 myeloid promoter in T cells. An enhancer located between the myeloid and epithelial APN/CD13 promoter increases the response of the promoter to the cell-cell contact induced expression of APN/CD13 in lymphocytes. Adhesion of lymphocytes to extracellular matrix did not result in increased promoter activity. The lymphocytic promoter response induced by direct cell-cell contact with SFCs is not affected by mutations of a proximal promoter element (nucleotides -48 to 35), which has a possible functional role in the basal APN/CD13 gene expression in lymphocytes. Upregulated peptidase-promoter activity via cell-cell contact shown in this study for the first time is discussed as a general mechanism in peptidase induction. PMID- 11029759 TI - Characterization of a nuclear factor that binds to AP1-like element in the rat p53 promoter during liver regeneration. AB - The transcription level of the rat p53 gene increases at 5-12 h in the regenerating liver after partial hepatectomy. It was previously reported that an activator protein 1 (AP1)-like element (-264--284) mediated the induced transcription of the rat p53 gene during liver regeneration. In this study, we characterize the protein binding to the AP1-like element by various methods. Oligonucleotide competition assays showed that the binding protein did not require AP1 consensus sequence. Therefore, the binding protein is not an AP1 family protein. Zn(2+) was required for maximum DNA-binding activity of the protein, suggesting that the binding protein contains zinc fingers. The binding protein was highly resistant to denaturant. Even 1.8 M urea did not eliminate the protein-DNA complexes. In addition, the binding protein was stable up to 55 degrees C. The protein-DNA complexes were abolished in the presence of 0.6 M NaCl and higher. Protease clipping assay showed that the protein had a protease resistant core DNA binding domain. These results provided new insights into the structure of the protein that binds to the AP1-like element of the p53 promoter during liver regeneration. PMID- 11029760 TI - Vitamin A as an enzyme that catalyzes the reduction of MTT to formazan by vitamin C. AB - The tetrazolium salt 3(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) is reduced to formazan by the succinate dehydrogenase system of active mitochondria, and hence, specifically used to assay for the viable cells, such as measurement of cell proliferation, cytotoxicity, and cell number. However, in the present study we have shown that some component specifically present in M199 but not in RPMI 1640 media can reduce MTT to formazan in the absence of a living system. Further study revealed that ascorbic acid reduced MTT to formazan, which was profoundly increased by a very small amount of retinol, whereas retinol alone had no effect. Oxidation of ascorbic acid by H(2)O(2) destroyed its ability to reduce MTT. The rate of MTT reduction was directly proportional to the concentration of MTT in the absence of retinol, but approached a zero-order state beyond a certain concentration of MTT in the presence of retinol. Furthermore, retinol remained unchanged after the completion of the reaction. Taken together, these results showed that retinol acts as a reductase that catalyzes the reduction of MTT to formazan using ascorbic acid as the cosubstrate (electron donor). PMID- 11029761 TI - Effects of calcium and magnesium on a 41-kDa serine-dependent protease possessing collagen-cleavage activity. AB - We report here the continued characterization of a 41-kDa protease expressed in the early stage of the sea urchin embryo. This protease was previously shown to possess both a gelatin-cleavage activity and an echinoderm-specific collagen cleavage activity. In the experiments reported here, we have explored the biochemical nature of this proteolytic activity. Pepstatin A (an acidic protease inhibitor), 1,10-phenanthroline (a metalloprotease inhibitor), and E-64 (a thiol protease inhibitor) were without effect on the gelatin-cleavage activity of the 41-kDa species. Using a gelatin substrate gel zymographic assay, the serine protease inhibitors phenylmethylsulfonyl fluoride and benzamide appeared to partially inhibit gelatin-cleavage activity. This result was confirmed in a quantitative gelatin-cleavage assay using the water soluble, serine protease inhibitor [4-(2-aminoethyl)benzenesulfonylfluoride]. The biochemical character of this protease was further explored by examining the effects of calcium and magnesium, the major divalent cations present in sea water, on the gelatin cleavage activity. Calcium and magnesium competed for binding to the 41-kDa collagenase/gelatinase, and prebound calcium was displaced by magnesium. Cleavage activity was inhibited by magnesium, and calcium protected the protease against this inhibition. These results identify calcium and magnesium as antagonistic agents that may regulate the proteolytic activity of the 41-kDa species. PMID- 11029762 TI - Type V collagen regulates the assembly of collagen fibrils in cultures of bovine vascular smooth muscle cells. AB - Vascular smooth muscle cells (SMCs), the major cellular constituent of the medial layer of an artery, synthesize the majority of connective tissue proteins, including fibrillar collagen types I, III, and V/XI. Proper collagen synthesis and deposition, which are important for the integrity of the arterial wall, require the antioxidant vitamin C. Vitamin C serves as cofactor for the enzymes prolyl and lysyl hydroxylase, which are responsible for the proper hydroxylation of collagen. Here, the role of type V collagen in the assembly of collagen fibrils in the extracellular matrix (ECM) of cultured vascular SMCs was investigated. Treatment of SMCs with vitamin C resulted in a dramatic induction in the levels of the cell-layer associated pepsin-resistant type V collagen, whereas only a minor induction in the levels of types I and III collagen was detected. Of note, the deposition of type V collagen was accompanied by the formation of striated collagen fibrils in the ECM. Immunohistochemistry demonstrated that type V collagen, but not type I collagen, became masked as collagen fibrils matured. Furthermore, the relative ratio of type V to type I collagen decreased as the ECM matured as a function of days in culture, and this decrease was accompanied by an increase in the diameter of collagen fibrils. Together these results suggest that the masking of type V collagen is caused by its internalization on continuous deposition of type I collagen on the exterior of the fibril. Furthermore, they suggest that type V collagen acts as framework for the initial assembly of collagen molecules into heterotypic fibrils, regulating the diameter and architecture of these fibrils. PMID- 11029763 TI - Expression screening of factors binding to the osteocalcin bone-specific promoter element OC box I: isolation of a novel osteoblast differentiation-specific factor. AB - Contributing to bone-specific expression of the osteocalcin gene is the promoter element OC Box I (-99 to -76), which binds both Hox proteins and another nonhomeodomain factor (designated OCBP for osteocalcin-box binding protein) (Hoffmann et al. [1996] J Cell Biochem 61:310-324). OCBP correlates with increased promoter activity and may, therefore, be important to development or maintenance of the osteoblast phenotype. To identify OCBP candidates, we used a multimerized OC Box I sequence to screen a gammagt11 cDNA expression library, constructed from the rat osteosarcoma osteoblastic ROS 17/2.8 cell line, for cDNA clones encoding factors that recognize this element. Mutant OC Box I sequences that do not bind OCBP and/or homeodomain proteins were used to counterscreen the cDNA isolates. Clones showing binding specificity were sequenced and further characterized for patterns of expression in different tissues and cell lines. Among the characterized nonhomeodomain-related isolates, we identified a nucleolin, a clone encoding rCAP2 that is related to myogenic differentiation and more importantly, a cDNA clone containing a previously uncharacterized gene that has been designated as a cell differentiation-related factor (DRF). DRF mRNA is highly expressed in ROS 17/2.8 cells and in a developmentally regulated pattern during osteoblast differentiation, being upregulated at the postproliferative maturation transition and coinciding with the induction of osteocalcin gene expression. The 7.7-kb transcript encoded by clone 44 is abundantly expressed in osteoblasts, but the mRNA was not present at detectable levels in bone and soft tissues by Northern blot analysis. However, related expressed sequence tags were recently reported in cDNA libraries of rat lung and mouse sympathetic ganglion. The identification of DRF represents a novel osteoblast differentiation-specific marker related to osteocalcin expression. The identification of DRF may further facilitate definition of gene regulatory mechanisms mediating the final stages of bone cell differentiation PMID- 11029764 TI - Effectiveness of mass screening for breast cancer in Japan. AB - BACKGROUND: Breast cancer screening has been conducted in Japan mainly by physical examination, the standard method for breast cancer screening according to the Law of Health Services for the Elderly. The purpose of this study was to evaluate the effectiveness of mass screening for breast cancer in Japan. METHODS: We calculated the average coverage-rates for breast cancer screening per year from 1986 to 1995 for women aged 30-69 years for all of the 3255 municipalities in Japan, selecting "high coverage-rate" municipalities with average coverage rates of 20%, 30%, 40% or more. Two municipalities were selected as "controls" for each high coverage-rate municipality, and were matched for population, National Health Insurance rate, and the age-adjusted death rate from cancer of the female breast in the period 1986-90. We compared the change in the age adjusted death rate from 1986-90 to 1991-95 of the high coverage-rate municipalities and the comparable controls. RESULTS: The percent reduction in the age-adjusted death rate from cancer of the breast in the high coverage-rate municipalities was statistically significantly greater than those in the controls. CONCLUSIONS: The results suggest that mass screening for female breast cancer, mainly by physical examination, contributed to the reduction of mortality from breast cancer. PMID- 11029765 TI - Interval cancers as an indicator of performance in breast screening. AB - BACKGROUND: The literature shows that breast screening performance varies between centers. Those who monitor the performance of screening programs must use appropriate methods to demonstrate quality. Analysis of interval cancer rates provides one such measure. This study analyses interval cancers over a ten-year period in a single center of the UK National Breast Screening Program, offering screening to a population of 45,000 women. METHODS: 108,948 screening examinations were undertaken between 1987 and 1996. Sequential interval cancer rates were studied and compared with the results of the Swedish Two-County trial. More detailed analysis of the first prevalence round was undertaken. RESULTS: Improvement in rates occurred after three years. Two-year time bands show some fluctuation in rates, indicating variation in performance. For one time period, rates equivalent to the Two County Trial were achieved, but this was not consistently maintained. Overall rates equate to performance reported elsewhere in the UK (25% in the first year after screening, 48% in the second year after screening and 48% in the third year after screening, equivalent Two County rates are 17%, 32% and 57%). The prevalence round analysis shows that undiagnosed cancers arose after the film reading stage in 88% of interval cases. The team was able to distinguish minor signs from significant lesions effectively. Assessment procedures were effective once the patient had been recalled. CONCLUSION: Interval cancer rates fell initially but reached a plateau. Recall procedures were effective in making the diagnosis, and cases were missed at the film reading stage. Consistency of performance may be important in achieving low rates. PMID- 11029766 TI - Significance of alpha 9 beta 1 and alpha v beta 6 integrin expression in breast carcinoma. AB - BACKGROUND: Both alpha 9 beta 1 and alpha v beta 6 integrins have been newly identified from the tracheal epithelium of guinea pig. It has been pointed out that alpha 9 beta 1 functions as a receptor for tenascin-C and osteopontin. As for the ligands of alpha v beta 6, fibronectin and tenascin-C have been identified. It has not been ascertained whether alpha 9 beta 1 and alpha v beta 6 are expressed in normal breast tissue, benign breast lesion or breast carcinoma. METHODS: Immunohistochemical staining for alpha 9 beta 1 and alpha v beta 6 was performed in benign breast lesion and breast carcinoma specimens. Western blotting was carried out on 11 breast carcinoma cases. RESULTS: alpha 9 beta 1 was expressed in the cytoplasm of carcinoma cells in 23 of 90 cases (26%) and alpha v beta 6 in the membrane of carcinoma cells in 16 of 90 cases (18%). However, these findings of alpha 9 beta 1 and alpha v beta 6 did not correlate with any clinicopathological factors including the patients' age, tumor size, histological type of carcinoma, location of carcinoma cells and hormone receptor status. With regard to the histological grade of carcinoma, alpha v beta 6 and alpha 9 beta 1 expression did not statistically correlate, although no expression of alpha v beta 6 was observed in 14 cases of Grade I. On Western-blott analysis strong and weak bands consistent with alpha v beta 6 were noted in the membrane fraction extracted from breast carcinoma cells. On the other hand weak bands consistent with alpha 9 subunit were noted in the whole cell lysates of breast carcinoma cells and very weak or no bands consistent with alpha 9 subunit were noted in the membrane fraction extracted from the breast carcinoma cells. CONCLUSIONS: Significance of alpha 9 beta 1 and alpha v beta 6 integrins expression in breast carcinoma was still unknown on clinicopathological examination. The findings of Western blot analysis may indicate that the transportation system of glycoproteins such as integrins to the cell membrane of carcinoma cells is disturbed, although these integrins can be produced. PMID- 11029767 TI - Expression of estrogen receptor alpha exon 5 and 7 deletion variant in human breast cancers. AB - BACKGROUND: An exon deletion variant of estrogen receptor (ER) mRNA has been reported, as one of the possible mechanisms of loss of ER function. METHODS: We examined the expression of exons 3, 5, and 7 in ER alpha mRNA and the frequency of exon deletion variant expression in 64 cases of human breast cancers and in 8 non-cancerous breast tissues using reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Approximately the same amount of wild-type (wt) mRNA was detected in all the non-cancerous breast tissues. In cancers, expression of wild-type exon 3 (w3), exon 5 (w5), and exon 7 (w7) was detected in 93.5%, 93.5%, and 91.3% of ER alpha protein (pER) positive cases, respectively, and 27.8%, 38.9%, and 44.4% in negative cases, respectively (p < 0.0001, p = 0.0035, and p = 0.0002). Although the variants for exon 5 (d5) and 7 (d7) were detected in both non-cancerous and cancerous tissues respectively, the variant for exon 3 was not detected at all. Comparatively, the ratio of d5/w5 was significantly higher in pER positive and progesterone receptor protein (pPgR) negative cases. CONCLUSIONS: We suspect that the exon 5 deletion does not work as a dominant positive. PMID- 11029768 TI - Human papillomavirus type 33 DNA in breast cancer in Chinese. AB - BACKGROUND: The association between human papillomavirus (HPV) and anogenital tumors, especially cervical cancer, is well documented. However, it remains unclear whether there is also a correlation between HPV infection and human breast cancer. METHODS: We used PCR and Southern blot hybridization to analyze HPV-related DNA specimens from 32 cases of invasive ductal carcinoma operated upon in the Shanghai region of China. RESULTS: DNA derived from HPV33 was detected in 14 cases (43.8%). No HPV16 or HPV18 DNA was detected in any of the cases in this study. This is the first report demonstrating a correlation between HPV33 infection and breast cancer. CONCLUSIONS: Our results suggest that HPV33 infection may be involved in the pathogenesis of breast cancer in Chinese. PMID- 11029770 TI - Cytomorphometric differentiation of intraductal proliferative breast lesions. AB - BACKGROUND: Although cytologic examination has been an indispensable procedure for the diagnosis of various breast diseases, it is often difficult to make a precise diagnosis of intraductal proliferative breast lesions preoperatively. The present study attempts to clarify the differentiation of the lesions by the cytologic morphometric approach. METHODS: Cytologic specimens from 21 intraductal lesions, including nine ductal carcinomas in situ (DCIS), seven ductal hyperplasias (DH), and five papillomas were evaluated. Using a microscope connected to a computerized video system, the mean nuclear area, the perimeter, the form factor, the largest to smallest diameter ratio of the nuclei (LS ratio), and the coefficient of variation of the nuclear area (NACV) were measured and analyzed. RESULTS: The mean nuclear area and perimeter were significantly higher in the cases of DCIS than in DH (p < 0.01) and papilloma (p < 0.005). Similarly, DCIS had higher NACV values than the other groups (p < 0.05 and p < 0.005, respectively). There were no significant differences in form factor or LS ratio. CONCLUSIONS: The quantitative estimation of cytologic nuclear features is useful for preoperative differential diagnosis of intraductal proliferative lesions of the breast. PMID- 11029769 TI - Intratumoral pyrimidine nucleoside phosphorylase (PyNPase) activity predicts a selective effect of adjuvant 5'-deoxy-5-fluorouridine (5'DFUR) on breast cancer. AB - BACKGROUND: Pyrimidine nucleoside phosphorylase (PyNPase) is the enzyme that converts 5'-deoxy-5-fluorouracil (5'DFUR) to 5-fluorouracil (5FU). Its activity in cancer tissue may correlate with the selective antitumor activity of 5'DFUR in breast cancer. METHODS: Two hundred and sixteen T2 breast cancer patients were treated consecutively with surgery followed by 5'DFUR (600 mg/body/day) + tamoxifen (20 mg/body/day) for 2 years. PyNPase activity in breast cancer tissue, determined by high-performance liquid chromatography, ranged from 4.2-626.0 micrograms FU/mg protein/hr (mean +/- SD, 203.5 +/- 122.4), and the examined patients were divided into two groups: group A (high PyNPase group), cases with the PyNPase activity equal to or more than the mean value of 203.5 micrograms FU/mg protein/hr, and group B (low PyNPase group), cases with activity less than the mean value. RESULTS: Although there was no difference in relapse-free survival (RFS) between groups A and B, among node-positive patients (n = 83) those in group A tended to have a longer RFS. When divided into subgroups according to estrogen receptor (ER) status, among node-positive and ER-positive tumors (n = 49), the RFS was significantly better in group A than in group B (p < 0.05). CONCLUSION: Intratumoral PyNPase activity might be of use as a predictor of the effect of adjuvant 5'DFUR on breast cancer. PMID- 11029771 TI - Immunohistochemical analysis of GCDFP-15 and GCDFP-24 in mammary and non-mammary tissue. AB - BACKGROUND: Gross cystic disease fluid protein (GCDFP)-15, a major constituent protein in breast cysts, is known to be a marker of breast cancer, while the diagnostic value of GCDFP-24, a protein with a molecular weight of 24,000 daltons, has not been determined. The aim of this study was to elucidate the usefulness of GCDFP-24 for the differential diagnosis of breast cancer in combination with GCDFP-15 and to characterize the histologic features of GCDFP-24 positive breast cancer. METHODS: A total of 326 samples including non-neoplastic tissue and benign and malignant tumors from several anatomic sites were examined using commercially available monoclonal antibodies against GCDFP-15 and GCDFP-24. RESULTS: In non-mammary tissue, GCDFP-15 was detected in skin, salivary gland, bronchial gland, prostate and seminal vesicle, and GCDFP-24 was detected in apocrine glands and peripheral nerve. Thirty-seven (44.6%) and 22 (26.5%) samples of 83 breast cancers were positive for GCDFP-15 and -24, respectively. Combined assays of GCDFP-15 and -24 raised the positive rate to 50.6%. The markers were not detected in tumors originating from gastrointestinal tract, bronchopulmonary structures or the genitourinary system. Breast cancers positive for both GCDFP-15 and GCDFP-24 were of lower histologic grade according to Bloom & Richardson's scoring system (p < 0.05). CONCLUSION: Immunohistochemical analysis of GCDFP-24 in combination with GCDFP-15 expression was useful for definitive diagnosis of breast cancers, and the expression of these markers correlated with low grade breast cancer. PMID- 11029772 TI - Cosmetic results and complications after breast conserving therapy for early breast cancer. AB - BACKGROUND: The cosmetic and functional results of breast conserving therapy for early breast cancer were evaluated. These are important endpoints in the assessment of breast conserving therapy in addition to tumor control and survival. The factors suspected to influence cosmesis were also analyzed. METHODS: In 206 patients with stage I and II breast cancer treated by wide excision and axillary dissection followed by radiation therapy, the cosmetic results and complications were analyzed. The cosmetic outcome was assessed by a scoring method and breast retraction assessment (BRA). As complications, arm edema and restriction of shoulder movement and late skin reactions were analyzed. RESULTS: Of the 206 patients 92% showed an excellent to good cosmetic score before radiation therapy. The score deteriorated, but gradually improved and stabilized after 1 year. Eighty-one percent of the patients had an excellent to good cosmetic score at 3 years. The BRA of the 206 patients was 1.8 cm on average before radiation therapy. It increased to 2.3 cm after termination of radiation therapy, and did not change thereafter. Tumor size over 2 cm (p = 0.005) and tumors in the inner quadrant (p = 0.003) were factors which negatively affected the cosmetic score at 3 years. Tumor size over 2 cm (p = 0.003), tumors in the upper quadrant (p = 0.005), or a nipple-tumor distance of more than 2 cm (p = 0.01) were also negative factors for the BRA at 3 years. Arm edema, restriction of shoulder movement, and late skin reaction were generally mild, and were observed in 12%, 0% and 34% of patients at 3 years, respectively. CONCLUSIONS: The overall cosmetic results of breast conserving therapy are acceptable and the complication rate is low. Tumor characteristics, tumor size, location and nipple tumor distance are factors that affect cosmesis. PMID- 11029773 TI - Mammographic and clinicopathological features of mucinous carcinoma of the breast. AB - BACKGROUND: We attempted to improve the effectiveness of diagnostic techniques in mammographic imaging of mucinous carcinoma of the breast by defining the characteristics of mammographic images and investigating correlations between these images and various clinicopathological findings. METHODS: Clinicopathological investigations of 92 lesions in 90 cases of mucinous carcinoma of the breast were made. Mammography demonstrated 80 lesions with identical tumor shadow characteristics and these were divided into three patterns, circumscribed, indistinct and blended. Correlations between clinicopathological findings and mammographic images were investigated. RESULTS: Patients with mucinous carcinoma of the breast usually present with a palpable mass. The lymph node metastasis rate in this study was low and prognosis in the early postoperative period was satisfactory. On mammograms, the circumscribed pattern was the most frequent. The investigation of the correlation between histological sub-type and mammographic pattern showed a high percentage of pure type lesions exhibited in the circumscribed pattern while those of mixed histologic type often showed the indistinct pattern. Calcification frequency demonstrated on mammography was 75% for the indistinct and mixed patterns, and approximately 50% for the circumscribed pattern. A high rate of calcification seen outside the tumor shadow suggested a high frequency of invasion and the spread of cancer to neighboring tissues. The circumscribed pattern was least frequently associated with lymph node metastasis, followed by the indistinct and blended patterns in that order. CONCLUSION: Investigation of clinicopathological factors and mammographic findings in mucinous carcinoma of the breast suggests that mammography provides clinically valuable information for the treatment of this disease. These findings indicate the importance of careful mammographic observation at the time of diagnosis. PMID- 11029774 TI - A case of inflammatory breast cancer following augmentation mammoplasty with silicone gel implants. AB - A 54-year-old-woman who underwent augmentation mammoplasty with silicone gel implants 30 years previously, visited our hospital with complaints of bloody nipple discharge, redness and itching of her right breast. Cancer of the right breast was diagnosed by dynamic magnetic resonance imaging (MRI) examination with Gadolinium (Gd)-DTPA enhancement. Radical mastectomy was subsequently performed. The histopathological findings demonstrated scirrhous and inflammatory breast cancer with invasion of dermal lymphatics. PMID- 11029775 TI - A case of malignant fibrous histiocytoma after breast conserving therapy for breast cancer. AB - A 45-year-old woman with malignant fibrous histiocytoma (MFH) of the breast following breast conserving therapy (BCT) is described. She noticed a lump in her left breast 52 months after BCT for breast cancer. The lump was excised and nodular fasciitis was initially diagnosed. However, the tumor recurred locally 4 times in the next 18 months. MFH was finally diagnosed. This case is considered to be radiation-induced sarcoma. The risk of radiation-induced sarcoma after BCT seems to be very low, however careful follow-up is necessary. PMID- 11029776 TI - Malignant proliferating trichilemmal tumor in the skin over the breast: a case report. AB - A proliferating trichilemmal tumor is relatively uncommon. It is composed of multiple cysts consisting of squamous epithelium with trichilemmal keratinization without granular layer interposition. This lesion usually occurs in the scalp of elderly women. We describe a 67-year-old woman with a malignant proliferating trichilemmal tumor in the skin over the breast. We first misdiagnosed the disease as a primary squamous cell carcinoma of the breast with a metastatic lymph node in the axilla because of the disease site and our unfamiliarity with the disease. The patient underwent radical mastectomy with axillary dissection. Eight months postoperatively, a tumor appeared in her right axilla and progressively enlarged. We subsequently excised the tumor. She is healthy as of 8 months postoperatively. To the best of our knowledge, only one case of a proliferating trichilemmal tumor occurring in the skin over the breast has been reported. PMID- 11029777 TI - A case of metastatic breast cancer achieving complete response by combination therapy with 5'-deoxy-5-fluorouridine and cyclophosphamide. AB - Recently there have been several reports on the effectiveness of combination chemotherapy with 5'-deoxy-5-fluorouridine (5'-DFUR) and cyclophosphamide (CPA) for treating recurrent breast cancer. We report a case in which treatment of local recurrence and lung metastases responded was remarkably effective by this combination chemotherapy. A 45-year-old woman underwent modified radical mastectomy for left breast cancer (T2N0M0) three years previously. Involvement of a left supraclavicular lymph node (ScLN) and multiple lung metastases were revealed 2 years and 6 months after the operation. First, we opted for systemic endocrine therapy with local irradiation for the lung metastases and ScLN. However the lung metastases were found to have increased on chest x-ray 2 months after treatment, showing progressive disease (PD). A locally recurrent new lesion sized 2 x 2 cm had also developed. Thus, we changed the treatment to combination chemotherapy with 5'-DFUR 800 mg/body and oral CPA 100 mg/body once a day. The lung metastases had disappeared on chest x-ray and the local recurrence was not palpable after 7 weeks of the new treatment. Thymidine phosphorylase (dThdPase) immunostaining of the primary tumor was strongly positive in almost all cancer cells. We discuss the mechanism of the increased efficacy of combination chemotherapy with 5'-DFUR and CPA. PMID- 11029778 TI - Dye- and gamma probe-guided sentinel lymph node biopsy in breast cancer patients: using patent blue dye and technetium-99m-labeled human serum albumin. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy is a promising method for the diagnosis of the axillary nodal status. We examined the availability of the SLN biopsy using two mapping procedures: the dye- and gamma probe-guided method, and preoperative lymphoscintigraphy by gamma camera imaging. METHODS: We enrolled 48 patients with breast cancer. Technetium-99m-labeled human serum albumin was injected into the subdermal tissue above the primary tumor or biopsy cavity, and preoperative gamma camera imaging was performed. After induction of general anesthesia, patent blue dye was injected into the peritumoral area prior to the surgical procedure. A handheld gamma-detection probe was used to assist in SLN detection. Careful dissection was performed to identify blue-stained afferent lymphatic vessels and nodes. An SLN was defined as any blue and/or radioactive node, and was excised. After SLN biopsy, axillary lymph node dissection of level I, II, and III was completed, in order to confirm the diagnostic ability of the SLN biopsy. RESULTS: Intraoperative SLN identification of axillary lesions was successful in 43 of 48 patients (90%). The dye- and gamma probe-guided method was successful in 25 patients (52%), the dye-guided method alone succeeded in 11 patients (23%), and the gamma probe-guided method alone succeeded in 7 patients (15%). Preoperative lymphoscintigraphy revealed axillary focal accumulations in 29 of 48 patients (60%). All patients who underwent successful preoperative SLN identification by lymphoscintigraphy had successful intraoperative SLN identification. A diagnostic accuracy of 95%, a sensitivity of 89%, and a specificity of 100% were achieved in the diagnosis of axillary metastasis. Internal mammary SLNs were identified in four patients intraoperatively, but we could not detect cancer metastasis in the internal mammary SLNs. CONCLUSIONS: The dye-guided and gamma probe-guided methods were complementary. Preoperative lymphoscintigraphy was useful to predict intraoperative SLN identification. Further study is necessary to assess the role of SLN biopsy of the internal mammary lymph nodes. PMID- 11029779 TI - Indications and technique of sentinel lymph node biopsy in breast cancer using 99m-technetium labeled tin colloids. AB - BACKGROUND: The status of the sentinel lymph node (SLN) can reflect the status of other lymph nodes in breast cancer. The efficacy of dye injection and radiolabeled tin colloids for the accurate identification of the SLN was investigated. The indications for SLN biopsy for determining clinical nodal status were also investigated. METHODS: A total of 108 patients with breast cancers less than 5 cm were enrolled. Ninety-six patients were clinically node negative and 12 were node positive. About 2 hours before surgery, 1 to 2.5 ml of 99m-technetium-labeled tin colloid was injected around the tumor. Just before the operation, dye was also injected into the tissue surrounding the tumor. Six clinically node negative patients were omitted from the dye-injection process. The SLN was identified as a lymph node with extremely high radioactivity using a gamma probe or a gamma counter. Complete axillary dissection was performed and the metastatic status investigated by hematoxylin and eosin staining. RESULTS: In clinically node negative patients undergoing dye-injection, the SLN was identified in 89 of 90 patients (98.9%), and there was only one patient with lymph node metastasis outside the SLN. However, in clinically node positive patients undergoing dye-injection, the identification rate of the SLN was 66.7% (8 of 12 patients) and there was one patient with lymph node metastasis outside the SLN (12.5%). Without dye-injection, the SLN could be detected in 4 of 6 patients (66.7%). CONCLUSIONS: Successful identification of the SLN with tin colloid requires concomitant dye-injection and candidates for SLN biopsy should be restricted to clinically node negative cases. PMID- 11029781 TI - Multidisciplinary approach to selective sentinel lymph node mapping in breast cancer. AB - Although the role of axillary lymph node dissection is controversial with respect to survival benefits, its role as a staging procedure has been well established since nodal involvement is the most reliable prognostic indicator for patients with breast cancer. Selective sentinel lymph node (SLN) dissection is gaining acceptance as a useful staging procedure because it is minimally invasive and spares approximately 70-80% of the patients a more extensive axillary lymph node dissection. The evolving techniques for selective SLN dissection using blue dye and radiotracer methods are reviewed in this article. Based on the classic definition of the breast lymphatic drainage and recently published articles addressing the issue of peritumoral and intradermal injections, a possible new and simplified approach using intradermal injection may identify the axillary SLN more quickly and reliably. This article emphasizes the importance of a multidisciplinary approach in the identification of SLNs by preoperative lymphoscintigraphy performed by expert nuclear medicine physicians, the intraoperative mapping and harvesting of SLNs by well trained surgeons and the meticulous examination of SLNs by experienced pathologists. Therefore, to achieve the highest rate of accuracy regarding SLN status, it is imperative that a multidisciplinary team with close communication and cooperation be formed. The clinical significance of SLNs will be determined by results from follow-up and clinical trials. PMID- 11029780 TI - Comparative efficacy of positron emission tomography and ultrasonography in preoperative evaluation of axillary lymph node metastases in breast cancer. AB - PURPOSE: In primary breast cancer, axillary nodal status is the most powerful predictive factor of recurrence. However, axillary lymph node dissection may cause surgical complications. If preoperative evaluation of axillary lymph node metastases is possible, unnecessary axillary lymph node dissections can be avoided. The purpose of this study was to evaluate the efficacy of positron emission tomography (PET) on detection of axillary lymph node metastases in breast cancer. METHODS: PET scans of the axilla were obtained in 32 patients with primary breast cancer. All patients fasted for at least 4 hours before the examination. After transmission scans for attenuation correction were performed, emission scans after intravenous injection of 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) were obtained. RESULTS: Overall accuracy of PET alone, ultrasonography alone, and in combination in the detection of axillary metastases were 82%, 79%, and 85% respectively. CONCLUSION: There were no significant differences between PET, ultrasonography, and PET in combination with ultrasonography regarding sensitivity, specificity and accuracy in the detection of axillary metastases. PMID- 11029782 TI - Clinical and pathologic factors predicting axillary lymph node involvement in breast cancer. AB - The diagnosis of axillary disease remains a challenge in the management of breast cancer and is a subject of controversy. In 1998, the Japanese Breast Cancer Society conducted a study assessing axillary lymph node involvement in breast cancer. The study included (a) clinical assessment by pre-operative imaging modalities, (b) histologic assessment for peritumoral lymphatic invasion, (c) biologic assessment by gelatinolytic activity using film in situ zymography, and (d) sentinel lymph node (SLN) biopsy. Clinical assessments by CT, PET, and US as well as biologic assessment were limited in their ability to detect axillary lymph node disease, although these imaging techniques may be useful to exclude node-positive patients from the need for SLN biopsy. Histologic assessment for peritumoral lymphatic invasion was useful, particularly for detecting false negative cases by SLN biopsy. Nevertheless, the utility of SLN biopsy in assessing axillary nodal status was confirmed. Axillary lymph node dissection (ALND) can be avoided in patients with a small tumor and a negative SLN. However, further studies will be required to investigate the value of SLN biopsy for predicting regional control and survival before it can replace routine ALND as the optimal staging procedure for operable breast cancer. PMID- 11029783 TI - Regulation of interleukin-6 secretion from breast cancer cells and its clinical implications. AB - Interleukin (IL)-6 may play possible roles in the proliferation and metastasis of cancer cells, in the development of osteolysis and humoral hypercalcemia, and in the regulation of estrogen production in breast cancer tissues. IL-6 is also suggested to be a cachectic factor in cancer patients. A decrease in serum IL-6 levels induced by medroxyprogesterone acetate (MPA) has been reported to correlate with a reversion of body weight loss in patients with advanced breast cancer. To elucidate the mechanisms of action of the anti-cachectic effect of MPA its effects on IL-6 secretion from the KPL-4 cell line, the first human breast cancer cell line to secrete IL-6 and to induce cachexia, were explored. It has been suggested that an inhibitory effect of MPA on IL-6 secretion from breast cancer cells causes the anti-cachectic effect of MPA. Our other studies have revealed that 5'-fluorouridine (5'-DFUR) inhibits the growth of KPL-4 tumors and decreases IL-6 levels in both serum and tumor tissues. Decreasing serum IL-6 levels resulted in alleviation of body weight loss. Docetaxel increased IL-6 levels in both serum and KPL-4 tumors, but combined treatment with docetaxel and 5'-DFUR resulted not only in a potent antitumor effect but also in a drastic decrease of serum IL-6 levels. In the present paper the possible roles of IL-6 in the development and progression of breast cancer are reviewed, and the regulatory mechanisms of IL-6 secretion from breast cancer cells and the possible clinical implications of decreasing IL-6 secretion by therapeutic agents are discussed. PMID- 11029784 TI - Medroxyprogesterone acetate and cancer cachexia: interleukin-6 involvement. AB - Cancer cachexia is a significant problem facing both patients and physicians. Many interventions have been tried in an attempt to remedy undernutrition in cancer patients. However, there is no convincing evidence that enteral/parenteral nutrition or the use of anabolic steroids is of any benefit in patients with cachexia. A recent prospective study revealed that oral medroxyprogesterone acetate (MPA) treatment reduces serum levels of interleukin (IL)-6, an important mediator of cancer cachexia, in patients with metastatic breast carcinoma regardless of response to the therapy. A decrease in serum IL-6 levels was well associated with subjective improvement in patients with metastatic breast carcinoma. Furthermore, clinically attainable concentrations of MPA can inhibit the growth of some human pancreatic carcinoma cells by inducing apoptosis in association with the phosphorylation of bcl-2. These results suggest that this agent may contribute to improved quality of life in patients with various cancers. PMID- 11029785 TI - Comparison of five different antibodies in the immunohistochemical assay of estrogen receptor alpha in human breast cancer. AB - BACKGROUND: Estrogen receptor alpha (ER) expression is the best prognostic and predictive factor of hormone dependency of human breast cancers. Unlike enzyme immunoassay (EIA), which has been widely used to evaluate ER status in breast cancer, immunohistochemical assay (IHC) can detect ER in a small amounts of tissue with detailed localization. Although there is a sufficient number of ER antibodies against various regions of the protein, the reliability of IHC staining is only well understood for a few. IHC and EIA for the evaluation of the ER status of human breast cancer, therefore, should be compared using the same breast cancer tissues. METHODS: Five different ER antibodies (1D-5, C-314, G-20, C-311 and HC-20) that identify different amino acid sequences were used. The evaluation of ER status by IHC using these antibodies was compared with EIA concomitantly in 97 primary human breast cancer tissues RESULTS: The positivity rate for EIA was 68%. That of IHC for antibodies 1D-5, C-314, G-20, C-311 and HC 20 was 50.5%, 47.4%, 46.4%, 44.3% and 57.7%, respectively. The concordance between EIA was 76.3% for 1D-5 and 77.3% for HC-20, which is statistically highly significant (p<0.0001); Other antibodies were not. CONCLUSIONS: HC-20 is most suitable in the evaluation of the ER status of human breast cancers using the IHC method. Although antibody 1D-5 is also available, C-314, G20 and C-311 are unreliable in such an evaluation. PMID- 11029786 TI - Skin reactions after breast-conserving therapy and prediction of late complications using physiological functions. AB - BACKGROUND: The temperature of the skin remains elevated long after breast conserving treatment with irradiation, perhaps because evaporative cooling is impaired. We investigated physiological changes of the irradiated skin and reevaluated the radiosensitivity of sweat glands on a functional basis to determine whether severe complications can be predicted. METHODS: Breast and axillary skin temperatures were measured with thermography and sweat production in response to local thermal stimuli was measured on the basis of changes in electrical skin resistance with a bridge circuit in 45 women before, during, and after breast irradiation for breast cancer. RESULTS: Breast and axillary skin temperatures were significantly increased after irradiation. In response to cutaneous thermal stimuli, the electric skin resistance of nonirradiated areas decreased significantly because of sweating, but that of irradiated areas was unchanged. CONCLUSION: Impairment of sweating may play an important role in skin damage after irradiation. Although glandular tissue is not usually radiosensitive, the results of our functional assessment suggest that sweat glands are more radiosensitive than expected. PMID- 11029787 TI - Readministration of tamoxifen after adjuvant therapy for recurrent breast cancer. AB - BACKGROUND: Previous series concerning tamoxifen (TAM) rechallenge did not obtain satisfactory results. Using stricter criteria, we now assess the usefulness of readministration of TAM as an initial therapy for patients with recurrent breast cancer. METHOD: The eligibility criteria were postmenopausal, estrogen receptor (ER) positive or unknown, at least 12 months of adjuvant TAM, a 6-month or longer drug-free period and no previous therapy after recurrence. A total of 10 patients were enrolled. TAM was administered in daily doses of 20 or 30 mg. RESULTS: The mean age of the patients at the time of recurrence was 64.8 years. The receptor status was positive in 8 patients and unknown in 2. The median disease-free interval (DFI) after mastectomy was 71.7 months. A complete response was observed in one patient, a partial response in 6, stable disease in 2, and progression in one. The response rate was thus 70%, with an additional two patients showing no progression over 6 months. Although only one patient with a DFI of less than 48 months showed a positive response, all patients with a DFI longer than 48 months showed a clinical response. The duration of response was less than 12 months in 3 patients and longer in 4. CONCLUSION: The post-adjuvant readministration of tamoxifen is a useful first choice therapy for postmenopausal recurrent breast cancer patients with positive ER and longer DFI. PMID- 11029788 TI - Seven cases of breast cancer recurrence limited to the central nervous system without other visceral metastases. AB - We report 7 rare cases of recurrent breast cancers who presented with central nervous system (CNS) metastases as the initial relapse site without any other organ metastases. The average age of the patients at surgery was 42.6 years old of age (median 45:range 32-60), and 6 of the 7 cases (86%) were premenopausal. The mean disease-free period was 25.7 months (median 22, range 2-60 months). The primary tumors were all invasive ductal carcinomas. The estrogen receptor and progesterone receptor status of the 3 tumors available for study were all negative. The metastatic CNS lesions included the cerebrum (4 cases), cerebellum, cervical spinal cord, and meninges. In 6 out of these 7 cases (86%), the CNS metastasis was the initial recurrent lesion. Multidisciplinary treatments including surgery, radiotherapy and systemic or intrathecal chemotherapy were given. Although the mean survival time from clinical manifestations of the metastases of the 4 deceased patients was 20 months (median 20.5; range 6-33), one patient treated with surgery and radiotherapy is been still alive18 years later. These cases were also notable for the fact that the only metastatic site was in the CNS only during the entire clinical course, except for 2 cases, one with ocular adnexa metastasis, and the other with cervical lymph node metastasis. Premenopausal patients with negative hormone receptor status are more likely to develop this type of recurrence, regardless of the histological type. It is necessary to pay attention to neurological symptoms and signs during follow-up of breast cancer patients. PMID- 11029789 TI - A case of breast cancer with endocrine features. AB - Cancer with endocrine features rarely occurs in the breast. We report a case of breast cancer with endocrine features in an 80-year-old woman. The tumors were treated by wide excision and a postoperative histopathologic diagnosis of carcinoma of the breast with endocrine features was made. Immunostaining was positive for chromogranin A, neuron-specific enolase and synaptophysin, but weakly argyrophilic. Widespread metastases occurred two months postoperatively, and the patient died of breast cancer 6 months after the first treatment PMID- 11029790 TI - A case of primary squamous cell carcinoma of the breast with rapid progression. AB - Primary squamous cell carcinoma of the breast is a rare disease for which no definite treatment or prognosis has been established. This report describes our experience with one case of primary squamous cell carcinoma of the breast that ended in death after rapid progression. The patient was a 58 year old woman who visited our hospital complaining of a growing tumor and pain in the right breast. Ultrasonographically the tumor consisted of solid tumor and a large cyst. Squamous cell carcinoma was diagnosed by fine-needle aspiration cytology. Neoadjuvant CEF was given, but the tumor continued to grow, so CEF was discontinued after one course and modified radical mastectomy was performed subsequently. There was no evidence of metastasis to lymph nodes. About 5 months post operatively CT of the head and chest X-ray demonstrated metastatic lesions in the brain and lungs. Resection of the cerebral tumor was performed. About 1 month after the operation, however, a new metastatic brain tumor was found and the patient later died. PMID- 11029791 TI - Lymph node metastasis from breast cancer diagnosed by F-18 FDG whole-body PET (case report). AB - We report a 48-year-old woman with lymph node (LN) metastases from breast cancer in whom diagnosis was difficult by conventional imaging methods but easy by F-18 fluorodeoxyglucose (FDG) PET. She was given a diagnosis of right breast cancer and then underwent modified radical mastectomy in December 1997. During routine CT and US studies in September 1998, a questionable LN measuring 1 cm in diameter was detected in the right supraclavicular region, but a definitive diagnosis of this LN was difficult by these tests. The patient underwent FDG PET test, which showed strong nodular activity (SUV= 7.1) in the right supraclavicular region, indicating a metastatic lesion. Moreover, an unexpected second metastatic LN lesion (SUV= 5.5) in the medial aspect of the right axillary was diagnosed by PET. The PET results were confirmed by pathological examination of the surgical specimens. FDG PET is a useful tool to detect small malignant lesions that are not easily diagnosed by conventional imaging techniques such as CT and US. It is expected that PET test will be feasible at many institutions. PMID- 11029792 TI - Tubular adenoma of the breast in a 73-year-old woman. AB - A 73-year-old woman presenting with a right breast mass is described. The patient underwent lumpectomy under a diagnosis of breast cancer. However, histopathologically the surgical specimen was tubular adenoma of the breast. This is a rare benign tumor that is difficult to differentiate from breast cancer clinically, especially in elderly patients. We describe two reported cases of tubular adenoma in patients older than 65-years in Japan, as well as the present case. PMID- 11029793 TI - A case of breast cancer diagnosed by inguinal lymph node metastasis. AB - We describe a case of a 58-year-old woman with right inguinal lymph node swelling and a T1 tumor in the right breast. She was referred with an 18-month history of the former complaint and a six-month history of the latter. Excisional biopsy of the inguinal lymph node revealed breast cancer metastasis. Radiographical examination showed no metastases to the lungs, liver or bone. Modified radical mastectomy was performed. Histological examination revealed solid tubular carcinoma, PT2, PM (axillary lymph node metastases 4/16), stage IV. Estrogen and progesterone receptors were negative. Three cycles of postoperative cyclophosphamide, adriamycin and 5-fluorouracil (CAF) chemotherapy were given, and the right inguinal area was irradiated with 40 Gy. The patient complained of swelling in both legs three years after surgery. Computed tomography revealed marked lymph node swellings in the pelvic cavity. She died six months later. Inguinal lymph node metastasis from breast cancer is very rare, although distant lymph node metastasis in the cervix occurs frequently. This case should help clarify how breast cancer metastasizes to distant lymph nodes. PMID- 11029794 TI - Sudden hemorrhage of the breast caused by breast cancer: a case report and review of the literature. AB - A rare case of sudden hemorrhage caused by breast cancer is reported. A 71-year old woman noted bleeding from her left breast. Physical examination of the left breast showed a localized open cavity accompanied by bleeding and coagulation. The patient had no history of breast trauma or anticoagulation therapy. Incisional biopsy followed by histological examination resulted in a diagnosis of granulation tissue with no cancer cells present. Mammography and ultrasonography indicated probable breast cancer. As a result, a second incisional biopsy was performed, which suggested invasive ductal carcinoma without histological skin invasion. A modified radical mastectomy was performed under a diagnosis of stage II breast cancer. Breast cancer with sudden hemorrhage is rare. We review the literature and discuss the cause of this unusual manifestation. PMID- 11029795 TI - Interleukin-4 and breast cancer. AB - IL-4 is a pleiotropic cytokine produced by T lymphocytes which acts on various cells of such as T and B lymphocytes, monocytes, fibroblast, endothelial cells, macrophages and some others. IL-4 was originally described as a B cell growth factor, and now known to provide potent anti-tumor activity against various tumors, including breast cancer. IL-4 can induce apoptosis in cultured breast cancer cells. In addition, it has been clarified that IL-4 plays an important role in the regulation of estrogen synthesis enzymes including 17beta-HSD and 3beta-HSD. These findings imply that IL-4 is a key enzyme not only for Th2 type immune reactions but also for tumor cell growth itself in human breast cancer. PMID- 11029796 TI - Trends of IL-6 and IL-8 levels in patients with recurrent breast cancer: preliminary report. AB - BACKGROUND: We reported that IL-6 and IL-8 levels at the beginning of treatment are predictive indicators of response to therapy and prognosis of patients with recurrent breast cancer. The aim of this study was to investigate the trend of IL 6 and IL-8 levels in heavily pretreated patients with recurrent breast cancer. METHODS: Cytokine level trends in 12 patients heavily pretreated with anthracyclines were studied. Patients were divided into two groups according to the objective response. There were 5 partial response (PR)/no change (NC), and 7 progressive disease (PD) patients. Blood was taken every four weeks. IL-6 was measured by chemiluminescent enzyme immunoassay. IL-8 was measured by ELISA. RESULTS: The pretreatment level of IL-6 in the PR/NC group (11.0+/-2.1 pg/ml) was significantly lower than that (15.3+/-2.7 pg/ml) in the PD group. However, there was no difference in IL-8 level between the PR/NC group (12.5+/-5.5 pg/ml) and the PD group (11.5+/-1.1 pg/ml). IL-6 levels in the PR/NC group were maintained within normal levels or decreased to within normal levels after treatment, while levels of IL-6 in the PD group gradually increased until the time of patient death. A decrease in IL-8 level after treatment was observed in only one patient in the PR/NC group. Mild increase of IL-8 levels was observed in the PD group. CONCLUSION: Continuous elevation of IL-6 levels indicates poor prognosis in heavily pretreated patients with recurrent breast cancer. Combination therapy including agents that reduce IL-6 levels will be a new strategy for aggressively treating recurrent breast cancer. PMID- 11029797 TI - Audit of a one-stop breast clinic--revisited. AB - BACKGROUND: A one-stop diagnostic service has been available for women with symptomatic breast disease at St Bartholomew's Hospital for 5 years and was originally audited in May 1993. In re-auditing the one-stop service our aim was to see if our practice had improved following the original audit and to look at the impact which these changes in practice had made to the service offered to the patient. METHODS: A prospective audit of 4 consecutive clinics was undertaken in November 1997. A total of 300 patients (59 new and 241 follow up) were seen in clinic in this time. The primary outcome measure concerned the completeness of triple assessment in the 40 patients who required one stop investigations, including mammography, ultrasonography and fine-needle aspiration cytology. In addition, mean time to initial consultation and level of awareness of the one stop facility and its attendant time delay were measured. RESULTS: Of the 300 clinic attendees 40 (38 new, 2 follow-up) had one-stop investigations. As a result of the one-stop service being in practice, 36 patients (90%) had a definitive management decision made at their first outpatient visit. Of these 2 were symptomatic cancers, forming 5% of the workload. A total of 86% of the workload was benign. Four patients (10%) had equivocal results. The mean waiting time from designated appointment until surgical consultation was 36.7 minutes and was disappointingly unchanged from that of the previous audit. However this does not take into account the significant reduction in staffing levels which has occurred between the two periods of assessment. CONCLUSIONS: The initial audit identified a significant problem with time constraints, necessitating that a large number of patients with carcinomas return at a later date for further investigations. Booking only new patients at the beginning of clinic has provided a solution. Disappointingly, our figures do not show a significant improvement in mean waiting time compared with the previous audit, despite allowing GPs greater access of referral. Encouragingly, we have been able to maintain a similar standard of provision of care despite lower staffing levels and to implement the changes suggested by the original audit (thereby closing the audit loop). PMID- 11029798 TI - Lymphatic mapping for breast cancer: experience at The Ohio State University. AB - BACKGROUND: Though not yet the standard of care, lymphatic mapping is becoming more widely utilized by surgeons who care for women with breast cancer. The purpose of this study is to report the early experience of lymphatic mapping at a large NCI designated cancer center. METHODS: Beginning in 1997, selected newly diagnosed breast cancer patients at our institution have undergone lymphatic mapping. Blue dye and radiolabelled colloid were used as mapping agents. Patients were entered into a prospective database which recorded demographics, mapping characteristics and pathologic correlation. RESULTS: In total, 352 patients were entered into the study, and 312 (89%) had an identifiable sentinel lymph node at the time of definitive surgery. Eight surgeons contributed to the database, four of whom performed more than 30 lymphatic mapping procedures. 149 patients underwent complete axillary lymphadenectomy either as part of a validation study (68) or because of metastasis disease to the sentinel node (81). The false negative rate was 4%. The surgeon's experience with the procedure was the only independent predictor of the ability to localize a sentinel node. CONCLUSIONS: Sentinel lymphadenectomy at our institution is an accurate means of predicting the status of the draining nodal basin. Experience with the technique correlates with successful localization, but patient selection and an institutional commitment to the procedure are also critical for success. PMID- 11029799 TI - Evaluation of histopathological criteria for identifying node-negative breast cancer with high risk of early recurrence in the NSAS-BC protocol study. AB - BACKGROUND: The histopathological criteria for high-risk node-negative primary breast cancer stated in the National Surgical Adjuvant Study of Breast Cancer (NSAS-BC) protocol were used to grade a consecutive series of 488 cases at our hospital. METHODS: To validate the criteria retrospectively, we examined the histological features of node-negative primary breast cancers which showed early relapse within 2 years after surgical therapy. RESULTS: Early relapse occurred in 12 patients, distant metastases in 11, and local recurrence in one. Among 278 cases followed for up to 1.5 years or longer, early systemic relapse was detected in 10 (5.8%) of 172 higher-grade tumors (9 invasive ductal carcinomas of nuclear grade 3 and one invasive ductal carcinoma of nuclear grade 2) and one stromal cell sarcoma. Among the 115 low-risk tumors, only one case (0.9%) of invasive ductal carcinomas with nuclear grade 1 showed early local recurrence. Early relapse occurred in only one (1.5%) of 67 tumors with an invasive component of 1.0 cm but in 11 (5.2%) of 211 tumors with an invasive component of 1.1 cm. The recurrence rate increased to 9.3% (8/86) when tumor invasion was 2.1 cm. In 12 cancers showing recurrence, strand structure, large central acellular zones, and squamoid features were histologically observed in four, two, and three cases, respectively. The present results confirmed the reported tendency of correlation between strand pattern and bone metastasis, large central acellular zones and lung and brain metastasis, and squamoid features and lung metastasis. Synchronous bilateral and unilateral multiple cancers were characterized by lower nuclear grades. CONCLUSIONS: At our hospital, the criteria used in the NSAS-BC protocol were demonstrated to identify node-negative cancers with high risk of early recurrence at a hospital level. To further identify groups prone to recurrence, longer follow-up would be necessary. In addition, the histological criteria could be improved to correlate with patient outcome more accurately. PMID- 11029800 TI - Thymidine phosphorylase expression correlates with tumor differentiation and Bcl 2 in invasive breast cancer. AB - BACKGROUND: Angiogenesis plays an important role in the growth and metastasis of solid tumors. Several angiogenic factors have been identified, and thymidine phosphorylase (TP) is thought to be one such factor. To date, little information is available on the relationship between TP and other clinicopathological variables. METHODS: Formalin-fixed, paraffin-embedded materials from 116 primary breast carcinomas were used. The expression of TP, estrogen receptor, Bcl-2, Bax, p53, c-erbB-2 and MIB-1 was examined by immunohistochemical methods. RESULTS: Nuclear and/or cytoplasmic TP expression was observed in the neoplastic cells, and accentuation of TP was often present at the infiltrating tumor edge and intraductal spread region. Tumor cell TP expression was significantly inversely correlated with histological grade (p< 0.05) and positively correlated with Bcl-2 expression, but no association with other tumor variables was found. CONCLUSIONS: TP is associated with Bcl-2 expression and tumor differentiation in breast cancer. TP may be a new prognostic parameter for breast cancer. PMID- 11029801 TI - Significance of the parathyroid hormone-related protein expression in breast carcinoma. AB - BACKGROUND: Parathyroid hormone-related protein (PTHrP) is produced by various neoplasms and is known to be a causative factor of hypercalcemia of malignancy. It has also been suggested to act as a cytokine for tumor progression. The purpose of this study was to clarify the significance of PTHrP expression in breast carcinoma. METHODS: PTHrP expression was examined in 177 surgically resected breast carcinoma specimens by immunohistochemistry using a monoclonal antibody against the for PTHrP, The relationship of PTHrP expression with clinicopathological factors was analyzed and the clinical courses of the patients are reported. RESULTS: Positive PTHrP staining was detected in 113 ( 64%) of the breast tumors. Among the positive cases, 36 (32%) of the tumors clearly showed strong expression. When the PTHrP expression was divided into three categories, a significant positive relationship was found between PTHrP expression and histological grade of tumor. PTHrP expression was also significantly related to bone metastasis but the staining degree of PTHrP was not. The patients with positive PTHrP tended to have poor outcome in proportion to the staining degree. Univariate analysis demonstrated a significantly shorter overall survival for patients expressing PTHrP, and in multivariate analysis showed that PTHrP status and nodal status were associated with a significantly shorter overall survival. CONCLUSION: Our results suggest that PTHrP expression is not only correlated with bone metastasis but is also related to the progression of breast carcinoma, and that overexpression of PTHrP may be a potential prognostic factor for human breast carcinoma. PMID- 11029802 TI - Cytokines facilitate chemotactic motility of breast carcinoma cells. AB - BACKGROUND: Both growth and motility of various tumor cells have been shown to be influenced by surrounding cells such as lymphocytes, histiocytes and fibroblasts through various cytokines, growth factors and extracellular matrices. The role of cytokines and extracellular matrices produced by lymphocytes, histiocytes and fibroblasts on migration and invasion of breast carcinoma cells has not been fully investigated METHODS: We investigated the effect of hepatocyte growth factor (HGF), interleukin (IL)-6, IL-8, IL-11, soluble type IV collagen and soluble laminin on the migration of 3 human breast carcinoma cell lines, MDA-MB 231, MCF-7 and T-47D, using a cell culture insert and a biocoat matrigel invasion chamber to assess migration across a matrigel-coated polyethylene telephtalate membrane. RESULTS: HGF, IL-6, IL-11 and IL-8 induced significant migration of MDA MB-231 cells depending on the dose of each cytokine. However, type IV collagen and laminin inhibited migration of MDA-MB-231 cells. In contrast, IL-8 inhibited migration of MCF-7 cells and IL-6 induced significant migration of T-47D cells, while no other cytokine or extracellular matrix induced significant migration of MCF-7 and T-47D cells. Only HGF induced significant invasion of MDA-MB-231 cells depending on the dose. MCF-7 and T-47D cells did not invade in response to any of the cytokines and extracellular matrices tested. CONCLUSIONS: These results suggest the possibility that the potency of chemotaxis or chemoinvasion differs according to the breast carcinoma cell line and that various cytokines and extracellular matrices secreted by lymphocytes, histiocytes and fibroblasts in the stroma of breast carcinoma can affect the invasion of breast carcinoma cells. PMID- 11029803 TI - Determination of optimal radiation energy for different breast sizes using CT simulator [correction of simulatior] in tangential breast irradiation. AB - BACKGROUND: The purpose of this study is to determine and recommend the optimal radiation source according to breast size for tangential irradiation in breast conserving therapy. METHODS: Twenty-eight patients treated at our department from January 1994 to January 1996 were studied. The dose distribution within the irradiated breast was calculated using a (60)Co-gamma ray and 6 MV-X ray. Then we compared 3-D dose distributions of the (60)Co-gamma ray and 6 MV-X ray in different-sized breasts. Three parameters (breast volume, chest wall separation, and breast height) were adopted as representative of breast size. We also examined correlations among the three parameters. RESULTS: When the breast size was large (breast volume >400 cm(3), chest wall separation > 19.5 cm, or breast height > 6.5 cm), the average volume of normal tissue which received more than 110% of the isocenter dose ("hot spot") was significantly greater with the (60)Co gamma ray than with the 6 MV-X ray (p < 0.05). A similar result was obtained with regard to hot spots in the clinical target volume. The cold area that received less than 95% of the isocenter dose was greater using a 6 MV-X ray when the breast size was small (breast volume <200 cm(3), chest wall separation <17.5 cm, or breast height <5.0 cm). However, the difference was not significant. There was a significant correlation between breast volume and chest wall separation (r =0.849, p <0.001). Breast volume and breast height were also significantly correlated (r =0.813, p <0.001). CONCLUSIONS: Since breast volume and shape are different in each patient, the optimal energy should be selected for each case to obtain uniform dose distribution in breast-conserving therapy. Chest wall separation or breast height, which are measurable without a 3-D planning system, can substitute for breast volume as parameters for breast size. We recommend that the (60)Co-gamma ray not be used for treating large breasts, those with chest wall separation > or =19.5 cm or breast height > or =6.5 cm. PMID- 11029804 TI - Investigation of menstruation recovery after LH-RH agonist therapy for premenopausal patients with breast cancer. AB - BACKGROUND: Recently Luteinizing hormone-releasing hormone (LH-RH) agonist has been used for premenopausal patients with breast cancer as endocrine therapy. However, there is no consensus regarding recovery of menstruation after long-term treatment with LH-RH agonist. We investigated recovery of menstruation after this treatment. METHODS: We investigated 28 premenopausal patients with breast cancer who underwent operation at Chiba Cancer Center Hospital in 1995 or 1996. The patients were treated with LH-RH agonist (goserelin) for 24 months as an adjuvant therapy, and were observed for more than 6 months after the last goserelin depot. Ages ranged 31-55 years old from at the time of last treatment. We defined recovery of menstruation as regular menstruation occurring more than three times. RESULTS: There were 22 patients in the recovery group (range: 31-53 years, mean 45.1 years). There were 6 patients in the non-recovery group (range: 50-55 years, mean 52.2 years). The overall recovery rate was 78.6%. Recovery rate was 81.8% for the patients under 50 years, and 66.7% for over 51 years. We separated all patients into groups by age at 5 years intervals, and investigated the distribution of the recovery time. In the recovery group (22 patients), there were 15 patients (68.2%) who confirmed recovery of menstruation within 6 months. CONCLUSION: Our investigation suggested that the recovery of menstruation after this therapy would occur in those less than 50 years old, within 6 months from the last treatment. PMID- 11029805 TI - Quantitative ultrasound as a predictor of node metastases and prognosis in patients with breast cancer. AB - BACKGROUND: A retrospective study was performed to determine whether preoperative quantitative ultrasound assessment could predict axillary lymph node metastases and prognosis in patients with breast cancer. We focused on the presence of a halo, which is one of the features of breast cancer on ultrasound and represents reflections from the invading margin around infiltrating malignancies. METHODS: We evaluated ultrasonography from 187 infiltrating breast carcinoma patients with tumors 5 cm or less in greatest dimension (T1, T2). Using computer image analysis, the halo area (H) and the sum of the area of halo and internal echo (total tumor area (T)) were measured, and the ratio of halo to entire tumor (H/T, halo ratio) was calculated and compared with lymph node status and prognosis. RESULTS: The mean of the halo ratio was 0.38+/-0.13. Using the value of 0.42 as a cut-off, the high halo ratio group had significantly worse prognoses for both overall and disease-free survival at 49 months in median follow-up (p <0.001 and p <0.0005, respectively). The specificity of a high halo ratio in the T1 classification for predicting axillary node metastasis was 83.1%, with a negative predictive value of 86.8%. In patients with tumors 1.0 cm or smaller, the negative predictive value was 100%. In a multivariate analysis, halo ratio was an independent predictor of disease-free survival of breast carcinoma patients (p =0.0232). CONCLUSIONS: Preoperative quantitative ultrasound may be a useful non invasive method for predicting the presence of axillary lymph node metastases and prognosis in patients with primary breast cancer. PMID- 11029806 TI - Interventricular methotrexate therapy for carcinomatous meningitis due to breast cancer: a case with leukoencephalopathy. AB - A 46-year-old woman presented with paraplegia and severe lumbago. She had had a radical mastectomy for left breast cancer 10 years earlier, and 6 months prior to presentation she completed CMF chemotherapy for treatment of retroperitoneal metastasis. CT and MRI to identify potential causes of the paraplegia and lumbago showed leptomeningeal carcinomatosis due to dissemination from invasive recurrence of the retroperitoneal tumor. An Ommaya reservoir was inserted, and infusion of intrathecal methotrexate (MTX; 5 mg twice weekly) began. Her clinical symptoms improved after receiving 53 mg MTX. However, after receiving 83 mg MTX, the patient became dizzy from leukoencephalopathy. Although administration of prednisolone mostly resolved her symptom, the patient died 9 months after the diagnosis of carcinomatous meningitis. PMID- 11029807 TI - Interstitial brachytherapy for recurrent breast cancer using a high dose rate Ir 192 remote afterloading system: a report of two cases. AB - We employed interstitial brachytherapy using a high dose rate Ir-192 remote afterloading unit in two breast cancer patients with locoregional recurrence. In the first case, skin metastasis was treated, with favorable control of the infield tumor but subsequent persistent sequelae and multiple outfield metastases. This experience caused us to be cautious when choosing brachytherapy for the second case, in whom a solitary metastasis to an axillary lymph node was successfully treated. Although this method is still investigational, it may play a critical role in the treatment of locoregional recurrence resistant to other treatment modalities. PMID- 11029808 TI - A case report of advanced male breast cancer with an objective response to tamoxifen treatment. AB - A 70-year-old man presented with a firm tumor in his right breast first noticed eight years ago. The tumor had enlarged gradually and had produced an ulcer with bleeding. On physical examination, a huge tumor entirely occupied the right breast and extensively had infiltrated the chest wall. Chest X-ray and CT showed massive pleural effusion and multiple small nodular lesions in the lung. Invasive ductal carcinoma of the breast was diagnosed by incisional biopsy,confirming advanced breast cancer with lung metastases and bilateral pleural effusion(T4cN2M1, Stage IV). Because ER and PgR levels were 110 fmol/mg and 190 fmol/mg, respectively, and because his general condition was poor, we selected medical treatment with tamoxifen(TAM). Thirty-two weeks later, the tumor had showed pronounced reduction with scarring. The patient underwent local excision of the scar tissue. The quality of life of the patient was favorably improved and no severe adverse events were observed. The tumor in the chest wall recurred two months after the end of TAM treatment, possibly because the patient did not accept continuous TAM therapy. The patient died from complications of brain metastasis 32 months after the start of TAM treatment. We report a rare case of advanced male breast cancer and on the effectiveness of continuous TAM treatment. PMID- 11029809 TI - ThurCatD: a tool for analyzing ratings on an ordinal category scale. AB - An algorithm for analyzing ordinal scaling results is described. Frequency data on ordinal categories are modeled for unidimensional psychological attributes according to Thurstone's judgment scaling model. The algorithm applies maximum likelihood estimation of model parameters. The Cramer-Rao bounds of the standard errors of the estimated parameters are calculated, and a stress measure and a goodness-of-fit measure are supplied. PMID- 11029810 TI - Determining the number of factors to retain: a Windows-based FORTRAN-IMSL program for parallel analysis. AB - Parallel analysis (PA; Horn, 1965) is a technique for determining the number of factors to retain in exploratory factor analysis that has been shown to be superior to more widely known methods (Zwick & Velicer, 1986). Despite its merits, PA is not widely used in the psychological literature, probably because the method is unfamiliar and because modern, Windows-compatible software to perform PA is unavailable. We provide a FORTRAN-IMSL program for PA that runs on a PC under Windows; it is interactive and designed to suit the range of problems encountered in most psychological research. Furthermore, we provide sample output from the PA program in the form of tabled values that can be used to verify the program operation; or, they can be used either directly or with interpolation to meet specific needs of the researcher. PMID- 11029811 TI - SPSS and SAS programs for determining the number of components using parallel analysis and velicer's MAP test. AB - Popular statistical software packages do not have the proper procedures for determining the number of components in factor and principal components analyses. Parallel analysis and Velicer's minimum average partial (MAP) test are validated procedures, recommended widely by statisticians. However, many researchers continue to use alternative, simpler, but flawed procedures, such as the eigenvalues-greater-than-one rule. Use of the proper procedures might be increased if these procedures could be conducted within familiar software environments. This paper describes brief and efficient programs for using SPSS and SAS to conduct parallel analyses and the MAP test. PMID- 11029812 TI - Empirical distributions of correlations as a tool for scale reduction. AB - A Visual Basic program for generating a distribution of correlations as a tool for scale reduction is presented; the program is available free from the author and runs on IBM-compatible personal computers under the Windows 95/98/NT operating systems. The program generates a list of item combinations and correlations between the unit weighted sum of the items and a criterion score. The list can be used to locate item combinations that have high correlations with the original full-length test. An example application is presented to illustrate the use of the program. The compiled program and its source code can be downloaded from http:?personal.bgsu.edu/stanton/r_crunch/r_crun ch.zip. PMID- 11029813 TI - Comparing choices and variations in people and rats: two teaching experiments. AB - Two pairs of experiments enabled students to compare their own operant behaviors with those of rats. The students played computer games for points, and the rats pressed levers for food. The first pair of experiments showed that, under concurrent schedules of reinforcement, relative frequencies of choices between two alternatives increased linearly in rats and people as functions of relative frequencies of reinforcement, with similar biases and undermatching observed in both species. The second pair of experiments showed that behavioral variability was controlled by reinforcers contingent on variability, this again true for both species. These experiments helped demonstrate the relevance of animal operant research to an explanation of human operant behavior. PMID- 11029814 TI - FISH 3: a microworld for studying social dilemmas and resource management. AB - A Java-based microworld environment for studying resource management is described. FISH 3 may be used in research or the classroom to investigate commons dilemmas and resource dilemmas. The program uses ocean fishing as its metaphor; participants ("fishers") experience the metaphor through both graphics ("fish" may be seen in an ocean) and text (e.g., resource replenishment is translated as "spawning"). In either stand-alone or networked modes, either with all human fishers or a mixture of human and computer fishers, 15 parameters such as the number of resource units (fish), participants (fishers), and trials (seasons), payoff values, the rate and period of resource regeneration (spawning), harvesting greed by computer fishers, awareness of other harvesters' actions, uncertainty in the amount of the resource, operating costs, and whether the resource is visible to harvesters may be varied. PMID- 11029815 TI - Investigating moral intensity with the World-Wide Web: a look at participant reactions and a comparison of methods. AB - This paper reports the results of a vignette- and questionnaire-based research project over the World-Wide Web investigating the influence of moral intensity (MI) on decision making in a business context. A qualitative analysis of the feedback in terms of e-mail communications was used to provide insights into the reactions and responses of participants to both the research method and the topic of research. Implications are discussed, and some methodological recommendations are derived. Second, analysis of the quantitative results of the Web-based questionnaire administration indicated that three of the six MI components were particularly important determinants of several outcome variables. This pattern of results essentially replicated findings yielded by a previous mail administration of the survey, even though a smaller amount of variation in the outcome variables was accounted for. Neither occupational background nor the region of origin of participants measurably influenced the results. PMID- 11029816 TI - PROXIMITY: an update and expansion of the PROXTIME computer program. AB - A new program, called PROXIMITY, was created as a tool for proximity calculation and to update and expand upon Kirste and Monge's (1983) PROXTIME program. The purpose of PROXIMITY is to calculate the fluctuating proximity between individuals within organizations. PROXIMITY provides output for three types of relationships: (1) an overall organizational proximity, (2) pairwise proximity between individuals, and (3) individual proximity to multiple others. PROXIMITY also updates some of PROXTIME's features such as the computer platform, the type of data the program can handle, and the form of output available. PMID- 11029817 TI - Software for recording observational files. AB - We offer the new software Codex, written in Visual Basic 3.0. It is a tool adequate in observational methodology. Its fundamental objective is to record motor and verbal behavior using the data types proposed by Bakeman and Quera (1995, 1996), together with the field formats proposed by Hall (1963), Weick (1968), Hutt and Hutt (1974), and Anguera (1979). It is designed to allow for data interchange between specific programs in use in observational methodology (SDIS-GSEQ, The Observer, and Theme) and other general programs (spread sheets, statistics applications, word processing programs, sound cards, etc.). PMID- 11029818 TI - EthoLog 2.2: a tool for the transcription and timing of behavior observation sessions. AB - EthoLog is a tool that aids in the transcription and timing of behavior observation sessions--experimental or naturalistic, from video/audio tapes or registering real time. It was created with Visual Basic and runs on Windows (3.x/9x). The user types the key codes for the predefined behavioral categories, and EthoLog registers their sequence and timing and saves the resulting data in ASCII output files. A sequential analysis matrix can be generated from the sequential data. The output files may be edited, converted to plain text files for printing, or exported to a spreadsheet program, such as MS Excel, for further analyses. PMID- 11029819 TI - Motion pattern and posture: correctly assessed by calibrated accelerometers. AB - Basic motion patterns and posture can be distinguished by multichannel accelerometry, as recently shown. A refinement of this method appeared to be desirable to further increase its effectiveness, especially to distinguish walking and climbing stairs, and body rotation during sleep. Recordings were made of 31 subjects, according to a standard protocol comprising 13 motions and postures. This recording was repeated three times with appropriate permutation. Five uni-axial sensors and three sites of placement (sternum with three axes, right and left thigh) were selected. A hierarchical classification strategy used a standard protocol (i.e., individual reference patterns) to distinguish subtypes of moving behaviors and posture. The analysis method of the actometer signals reliably detected 13 different postural and activity conditions (only 3.2% misclassifications). A minimum set of sensors can be found for a given application; for example, a two-sensor configuration would clearly suffice to differentiate between four basic classes (sitting, standing, lying, moving) in ambulatory monitoring. PMID- 11029820 TI - A low-cost, accurate method of producing large quantities of digitally filtered images. AB - We demonstrate how to produce complex image transformations of bitmap files for vision experiments using the Cogimatic Vision Starter Kit (VSK) library of mathematical routines along with Visual Basic, C++, or the Delphi Pascal compiler. Implementing this system on an IBM-compatible PC running Windows 95, 98, or NT4 enables researchers to quickly and economically manipulate images for vision research. The VSK includes a simple stand-alone image-processing application. In addition, VSK has the ability to automate image transformations and to fully integrate image processing into new experimental software on the PC platform. PMID- 11029821 TI - A MATLAB toolbox for the analysis of articulatory data in the production of speech. AB - The goal of this paper is to present EMATOOLS, a set of scripts for displaying and annotating acoustic and articulatory data simultaneously in studies on speech production. These scripts were developed with the use of MATLAB, a multiplatform computing environment for numeric computation and visualization. The system is equipped with a mouse-driven graphical interface made up of a number of displays. This interface can be easily customized to speed up routine tasks. The scripts can also be used in a noninteractive way, as stand-alone MATLAB commands. Output data can be imported into any standard spreadsheet. EMATOOLS is freely available from www.lpl.univ-aix.fr/nguyen/ematools.html. PMID- 11029822 TI - Automatic disambiguation of morphosyntax in spoken language corpora. AB - The use of computer tools has led to major advances in the study of spoken language corpora. One area that has shown particular progress is the study of child language development. Although it is now easy to lexically tag every word in a spoken language corpus, one still has to choose between numerous ambiguous forms, especially with languages such as French or English, where more than 70% of words are ambiguous. Computational linguistics can now provide a fully automatic disambiguation of lexical tags. The tool presented here (POST) can tag and disambiguate a large text in a few seconds. This tool complements systems dealing with language transcription and suggests further theoretical developments in the assessment of the status of morphosyntax in spoken language corpora. The program currently works for French and English, but it can be easily adapted for use with other languages. The analysis and computation of a corpus produced by normal French children 2-4 years of age, as well as of a sample corpus produced by French SLI children, are given as examples. PMID- 11029823 TI - A Japanese logographic character frequency list for cognitive science research. AB - This paper describes a Japanese logographic character (kanji) frequency list, which is based on an analysis of the largest recently available corpus of Japanese words and characters. This corpus comprised a full year of morning and evening editions of a major newspaper, containing more than 23 million kanji characters and more than 4,000 different kanji characters. This paper lists the 3,000 most frequent kanji characters, as well as an analysis of kanji usage and correlations between the present list and previous Japanese frequency lists. The authors believe that the present list will help researchers more accurately and efficiently control the selection of kanji characters in cognitive science research and interpret related psycholinguistic data. PMID- 11029824 TI - The rate and the type of orthodontic tooth movement is influenced by bone turnover in a rat model. AB - The influence of bone metabolism on both the rate and the type of orthodontic tooth movement was investigated. A rat model in which high (n = 16) and low (n = 17) bone turnover was pharmacologically induced was used. A non-pharmacologically treated group (n = 19) served as the control. A mesially directed constant single force of 25 cN was applied to the upper left first molar for a period of 3 weeks. The study was performed as a split-mouth design, the contralateral side of each animal serving as its control. The displacement of the molar crown was measured with an electronic calliper, while changes in inclination of the teeth were measured from micro-CT scans of the excised maxillae. The bone turnover significantly affected the rate of tooth movement. In the case of high turnover, the rate of tooth movement was increased while it was reduced in the case of low turnover. A controlled mesial tipping in all three groups was observed, but the actual location of the centre of rotation seemed to be influenced by the metabolic state of the bone. Based on the results it can be concluded that deviations in bone turnover influence the response to orthodontic forces, and should be taken into consideration when planning orthodontic treatment in patients with metabolic bone disease or those on chronic medication influencing bone metabolism. PMID- 11029826 TI - Surgical mandibular advancement and changes in uvuloglossopharyngeal morphology and head posture: a short- and long-term cephalometric study in males. AB - The aim of the present study was to investigate, by means of an extensive cephalometric examination, the alterations which took place in hyoid bone position, head posture, position and morphology of the soft palate, and tongue and sagittal dimensions of the pharyngeal airway after mandibular advancement osteotomy for the correction of mandibular retrognathism. The sample consisted only of adult males who underwent mandibular advancement by bilateral sagittal ramus split osteotomy (BSRO) with rigid fixation. Profile cephalograms were obtained 1-3 days before surgery (20 subjects), and 6 months (20 subjects) and 3 years (19 subjects) after the surgery. Statistical evaluation was performed by paired Student's t-test and Pearson product moment correlation analysis. At the short-term follow-up, hyoid bone and vallecula assumed a more superior (AH perpendicular FH, AH perpendicular ML, AH perpendicular S, V perpendicular FH) and anterior position (AH-C3 Hor, V-C3), which was maintained at the long-term follow-up. The soft palate (NL/PM-U) became more upright at the short-term follow up. The tongue demonstrated a transient increase in height (H perpendicular VT) and a less upright position (VT/FH) at the long-term observation. In addition, a more upright cervical spine (OPT/HOR, CVT/HOR) was recorded at the long-term follow-up. The pharyngeal airway space at the level of the oropharynx (U-MPW) and the retroglossal space at the base of the tongue (PASmin) showed an increase in the sagittal dimension at the short-term follow-up. Significant widening at the PASmin level was sustained at the long-term follow-up, indicating that mandibular advancement osteotomy could increase airway patency and be a treatment approach for sleep apnoea in selected patients. PMID- 11029825 TI - A cephalometric comparison of subjects with snoring and obstructive sleep apnoea. AB - This prospective study analysed the upright lateral cephalometric radiographs of 115 dentate, Caucasian males. Forty-five subjects exhibited proven obstructive sleep apnoea (OSA), 46 were simple snorers, and the remaining 24 subjects, who had no history of respiratory disease and did not snore, acted as controls. Radiographs were traced and digitized, and comparisons were made of the dento skeletal, soft tissue, and oropharyngeal features of the three groups. Differences were also sought between the snoring and OSA subjects. Of the hard tissue measurements, only the cranial base angle and mandibular body length showed significant inter-group differences (P < 0.001 and P < 0.05, respectively). When the airway and associated structures were examined, both snorers and OSA subjects exhibited narrower airways, reduced oropharyngeal areas, shorter and thicker soft palates, and larger tongues than their control counterparts. Comparison of the two sleep disordered breathing groups showed no differences in any of the skeletal or dental variables examined. However in OSA subjects, the soft palate was larger and thicker (P < 0.05), both lingual and oropharyngeal areas were increased (P < 0.01 and P < 0.05, respectively) and the hyoid was further from the mandibular plane (P < 0.05). Thus, whilst the dento skeletal patterns of snorers resembled those of subjects with OSA, some differences in soft tissue and hyoid orientation were apparent. There was not, however, a recognizable gradation in size of the airway and its associated structures from control through snoring to OSA subjects. This suggests that there may be a cephalometrically recognizable predisposition towards the development of sleep disordered breathing, but that this is only one facet of the condition. PMID- 11029827 TI - Surgical mandibular setback and changes in uvuloglossopharyngeal morphology and head posture: a short- and long-term cephalometric study in males. AB - A detailed cephalometric analysis was conducted on a sample of 31 adult males who underwent correction of mandibular prognathism by mandibular setback osteotomy (BSRO) with rigid fixation to evaluate the changes in uvuloglossopharyngeal morphology, hyoid bone position and head posture. Lateral cephalograms were obtained 1-3 days prior to the operation and at standardized 6 months and 3 years post-operative follow-up. Statistical evaluation was performed by paired Student's t-test and Pearson product moment correlation analysis. Inferior position of the hyoid bone (AH perpendicular FH, AH perpendicular ML, AH perpendicular S) and valeculla (V perpendicular FH) was recorded at the 6-month follow-up, a transient finding as at 3 years almost complete recovery to their pre-surgical position was noted. No posterior displacement of the above structures (AH-C3 Hor, V-C3) was recorded. Soft palate length (pm-U) was increased and maintained at the long-term follow-up while its posture (NL/pm-U) became less upright. The tongue showed increased length (V-T) and sagittal area (TA) and a more upright posture (VT/FH) at the late follow-up. Increased contact length between tongue and the soft palate (CL) and less residual oropharyngeal area [area not occupied by soft tissues, (TA + SPA)/OPA] was found at the long term follow-up. Craniocervical agulation (NSL/OPT, NSL/CVT) was increased indicating cervical hyperflexion at the 3-year follow-up. Reduction of the sagittal dimension of the oropharyngeal airway space (U-MPW) appeared at the first follow-up and was sustained at the longest follow-up which, in conjunction with the decrease in residual oropharyngeal area, could raise questions regarding airway patency after mandibular setback osteotomy. PMID- 11029828 TI - Degree of cure of orthodontic adhesives with various polymerization initiation modes. AB - The purpose of this study was to estimate the degree of cure (DC) of a light cured, and a two- and a one-phase (no-mix) chemically-cured, as well as a dual cured commercially available orthodontic adhesive resin. Forty stainless steel brackets were divided into four groups of 10 brackets each, and the bracket bases were covered with a standardized volume of adhesive. They were then pressed firmly onto a yellowish background surface of 75 per cent reflectance covered with cellulose film to facilitate detachment of the system and recovery of the set material. The visible light- and dual-cured adhesives were photopolymerized by irradiation from the incisal and cervical edges of the bracket for 10 seconds each, while another group of ceramic brackets was used to assess the differential interference of transparent relative to opaque material in the DC. Micro-multiple internal reflectance Fourier transform infrared spectroscopy was employed for the estimation of the DC of the adhesives (alpha = 0.05, n = 10), utilizing a method proposed previously. The dual-cured product demonstrated the highest DC followed by the light-cured combined with the ceramic bracket, and the no-mix and the chemically-cured adhesives. The combination of the metallic bracket with the light-cured product resulted in a DC comparable with that of the chemically-cured material. PMID- 11029829 TI - Fluoride-releasing elastomerics--a prospective controlled clinical trial. AB - A prospective controlled clinical trial was undertaken to evaluate the effectiveness of stannous fluoride-releasing elastomeric modules (Fluor-I-Ties) and chain (Fluor-I-Chain) in the prevention of enamel decalcification during fixed appliance therapy. Forty-nine patients (782 teeth) were included in the experimental group, where the fluoride-releasing elastomerics were used. Forty five patients (740 teeth) who received non fluoride-releasing elastomerics formed the control group. All patients had their elastomerics replaced at each visit. Enamel decalcification incidence and distribution were recorded using an index by direct clinical observation. In the control group enamel decalcification occurred in 73 per cent of patients and in 26 per cent of all teeth. In the experimental group the corresponding incidence was 63 and 16 per cent, respectively. The overall reduction in score per tooth produced by the fluoride-releasing elastomerics was 49 per cent, a highly significant difference (P < 0.001). A significant difference was seen in all but the occlusal enamel zones. The majority (over 50 per cent) of lesions occurred gingivally. The teeth most severely affected were the maxillary lateral incisors and mandibular second premolars. There was no difference in treatment duration between groups. PMID- 11029830 TI - Influence of ceramic and stainless steel brackets on the notching of archwires during clinical treatment. AB - The surface topography of 100 clinically used archwires of stainless steel, beta , or nickel-titanium were investigated that had contacted either ceramic or stainless steel brackets. One group consisted of two sets: 60 wires with no treatment records accessed to bias analyses, and 40 wires for which extensive clinical records were available, half of which were used with ceramic or stainless steel brackets. A control group consisted of two sets: 30 unused wires comprised of five round and rectangular wires of each alloy, and four wires that were ligated and immediately removed from patients' mouths. After ultrasonic cleaning, each wire was inspected under an optical and/or a scanning electron microscope. Notches were categorized with regard to frequency, patterns, and severity, and mapped as a function of wire aspect (lingual, facial, and occlusal/gingival) and anatomical regions (molar, premolar, canine, and incisor). From these data the average severity of notch patterns and a notching index were derived. Although no recognizable defect patterns were observed in the control group, seven basic patterns were recognized for each wire cross-sectional shape in the clinically used wires. These wires appeared most damaged on their lingual aspect and least damaged on their facial aspect. With regard to anatomical regions, notching was prevalent in the anterior regions and sparse in the molar regions. The notch activity and the severity were nearly three times greater from ceramic brackets than from stainless steel brackets. Over one-third of all notches documented in ceramic bracket cases had severity numbers of 3 and penetrated at least one-quarter of each wire's dimension, However, over two thirds of all notches documented in stainless steel bracket cases had severity numbers of 1. From these tabulations a theory of notch formation was proposed in which vertical movement from tooth or wire during mastication caused fretting wear, and horizontal movement during orthodontic procedures such as space closure, tipping, or bodily movement caused sliding wear. PMID- 11029831 TI - [New antiplatelet agents]. PMID- 11029832 TI - [Clinical and tomographic aspects of hemorrhagic cerebrovascular disease associated with hypertensive crisis in adults under 50 years of age]. AB - The purpose of this study was to analyze both the clinical and tomographic aspects of the hemorrhagic cerebrovascular disease (HCd), associated with hypertensive crisis in adults under 50 years of age. Forty six patients, who were not under anticoagulant therapy, were not using illegal drugs, who had not a cerebral tumor disease, and who had neither arteriovenous malformations nor past traumatic episodes, were studied. Seventy eight percent of the patients had preexisted arterial hypertension, 30% of them had at least a previous emergency for a hypertensive crisis. Mortality for intracerebral hematoma (ICH) and for subarachnoid hemorrhage (SAH) was 21% and 23% respectively. In 68% of the cases, ICH was located in the deep structures of the brain. Asymmetric ventricular system, compression or the absence of mesencephalic cisterna were significantly associated (p > 0.01; p > 0.001 respectively) with higher mortality. There was not a significant difference between the deceased and the survivors in relation with their systolic and diastolic arterial pressure on admission to the emergency unit. A significant positive relation was found between the severity of the injury (percentage of patients with an Scale Coma Glasgow < or = 8 points) and the mortality percentage for the type of HCd (r = 0.81 for ICH; p < 0.001, r = 0.98 for SAH; p < 0.001). Age and a low Scale Coma Glasgow score on the admission, represent unfavorable prognostic factors. Due to the different criteria used to evaluate the tomographic characteristics of intracerebral hematomas, comparisons of the present results with other findings can be difficult. PMID- 11029833 TI - Computerized brain tomography in patients with Schizophrenia. AB - We carried out brain tomography of 27 patients with schizophrenia and 17 normal control subjects. DSM-IV criteria were used for diagnosis. The parameters studied were: brain volume, lateral and third ventricles, brain surface and brain indexes. Previously the patients were evaluated with the following clinical scales: Positive and Negative Syndrome Scale (PANSS), Brief Psychiatric Rating Scale (BPRS), Global Assessment of Functioning Scale (GAF) and the GRAFFAR scale to evaluate cultural and socioeconomical levels. We detected an increase in the right and left lateral ventricles: brain ratios, an increase in the width of Sylvian fissures and in the third ventricle index. We did not find differences that could be related to sex or to a familial history of mental disease. No differences were observed in the group receiving medication when compared with the group of untreated patients. PMID- 11029834 TI - Clinical trial with Secnidazole in a single dose in Venezuelan children infected by Giardia intestinalis. AB - The aim of this work was to evaluate in an open, noncomparative study the use of secnidazole in oral suspension given to Venezuelan children infected with Giardia intestinalis, from a community in Carapita, a slum area in Caracas. Seventy children from 2 to 11 years old (38 males and 32 females) were treated with a single oral dose of secnidazole (30 mg/Kg of body weight), after clinical and parasitological evaluation to make the diagnosis of active giardiasis. The effectiveness of treatment was determined by clinical examination and parasitological evaluation of feces samples 15 days after treatment. The results showed 95% of clinical cure with a significant decrease of the frequency of gastrointestinal symptoms. The parasitological cure was 98%, there were 4 failures at the end of treatment. Side effects observed after treatment were of mild intensity, lasting only few hours. These results show that a simple dose of secnidazole in an oral suspension is an effective, safe and well tolerated treatment for giardiasis in children and that this drug may be used as a mass treatment in risk populations. PMID- 11029835 TI - [Prions, infections and confusions in the "transmissible" spongiform encephalopathies. The other evidence-based science. III. Review]. AB - There are some neurological disorders with a pathological hallmark called spongiosis which include Creutzfeld-Jakob disease and its new variant, the Gertsmann-Straussler-Scheinker Syndrome and the Fatal Familial Insomnia in humans; and Scrapie and Bovine Spongiform Encephalopathy, among others, in animals. The etiological agent has been considered either transmissible or hereditary or both. Curiously, this agent has no nucleic acids, is impossible to filter, is resistant to inactivation by chemical means, has not been cultured and is unobservable at electron microscopy. All of these facts have led to some researches to claim that these agents are similar to viruses appearing in computers. However, after almost fifty years of research, is still not possible to explain why and how such elements produce the diseases commented about. On the contrary, during these years have been possible to know that these entities called slow viral infections, transmissible amyloidosis, transmissible dementia, transmissible spongiform encephalopathies or prion diseases appear in individuals with genetical predispositions exposed to several worldwide immunological stressors. The possibility that prions are the consequence and not the cause of these diseases in animals and man is day by day more reliable, and supports the suggestion that a systematic intoxication due to pesticides as well as mycotoxin ingestion, produced mainly by different molds such as Aspergillus, Penicillium or Fusarium, seem to be the true etiology of these neurodegenerative disorders. PMID- 11029836 TI - Providing drug information in remote regions of Japan. PMID- 11029837 TI - Prepackaging would prevent contamination. PMID- 11029838 TI - A flawed example. PMID- 11029839 TI - The pharmaceutical industry, 1852-1902. PMID- 11029840 TI - APhA drug treatment protocols: management of pediatric acute Otitis media. PMID- 11029841 TI - Frequency and characteristics of cognitive services provided in response to a financial incentive. AB - OBJECTIVE: To determine the effects of a financial incentive on the number and types of cognitive services (CS) provided by community pharmacies to Medicaid recipients in the State of Washington. DESIGN: Prospective randomized trial. CS were reported using a problem-intervention-result coding system over a 20-month period. SETTING AND SUBJECTS: Pharmacists practicing in 110 study (financial incentive) and 90 control community pharmacies. RESULTS: Study pharmacists documented an average of 1.59 CS interventions per 100 prescriptions over a 20 month period, significantly more than controls, who documented an average of 0.67 interventions (P < .05) per 100 prescriptions. One-half (48.4%) of all CS were for patient-related problems, 32.6% were for drug-related problems, 17.6% were for prescription-related problems, and 1.4% were for other problems that did not involve drug therapy. A change in drug therapy occurred as a result of 28% of all CS documented in this demonstration. Changes were rarely (2.4%) due to generic or therapeutic substitution and almost always (90%) followed communication with the prescriber. The average self-reported time to perform CS was 7.5 minutes; 75% of interventions were < or = 6 minutes. Considerable differences existed between study and control groups in the types of problems identified, intervention activities performed, and results of interventions. CONCLUSION: A financial incentive was associated with significantly more, and different types of, CS performed by pharmacists. PMID- 11029842 TI - Patients' willingness to pay for pharmaceutical care. AB - OBJECTIVE: To determine the level at which patients receive pharmaceutical care services and their willingness to pay for comprehensive pharmaceutical care services. DESIGN: A mail survey was sent to 2,500 adults in the United States. SETTING: Surveys were mailed to subjects' homes. PATIENTS OR OTHER PARTICIPANTS: Subjects were randomly selected from a marketing database that included representation from each of the 50 states of the United States. INTERVENTION(S): The survey provided a description of comprehensive pharmaceutical care, and survey items asked about the level of care subjects were receiving and their willingness to pay for these services. MAIN OUTCOME MEASURES: Level of various pharmacy services subjects reported receiving, and the dollar amount subjects were willing to pay for comprehensive pharmaceutical care. RESULTS: The majority of the subjects were not receiving pharmaceutical care services. The average amount all respondents were willing to pay for these services was $13 for a one time consultation and $28 for this plus 1 year of monitoring. Looking only at those respondents willing to pay (56%), the means rise to $23 and $50, respectively. CONCLUSION: A majority of patients are willing to pay for pharmaceutical care services, even if they are not now receiving this level of care. Direct payment from patients who recognize the therapeutic benefits of pharmaceutical care may be a more viable option than is generally believed, at least until the profession can prove pharmaceutical care's utility and cost effectiveness to third party payers. PMID- 11029843 TI - Physicians' perceptions of communication with and responsibilities of pharmacists. AB - OBJECTIVE: To understand physicians' perceptions of (1) their communication with pharmacists, (2) pharmacists' professional duties, and (3) the degree of responsibility with which pharmacists perform these tasks. DESIGN: Structured interviews with convenience sample of physicians in phase 1; written survey mailed to convenience sample of physicians in phase 2. PARTICIPANTS: Six physicians in Wyoming in phase 1, and 313 primary care physicians who were members of the Wyoming Medical Society in phase 2. MAIN OUTCOME MEASURES: Physicians' attitudes and experiences related to their interactions with pharmacists and their perceptions about pharmacists' responsibilities. RESULTS: Usable surveys were returned by 176 physicians (response rate = 59.1%). Age ranged from 27 to 86 years (mean +/- SD = 43.5 +/- 10.4 years), 79.5% were men, and 50.6% were in family practice. Almost 25% had personal contact with pharmacists regarding patients' medications four or more times daily, but 20.6% rarely had this type of contact. Pharmacists contacted physicians' offices regarding prescription refills frequently, with 28.7% reporting 10 or more contacts daily. For 79.2% of respondents, an office nurse had the most contact with pharmacists. Respondents were most comfortable with pharmacists' responsibilities of catching prescription errors (88.0%), providing patient education (65.1%), suggesting nonprescription medications (63.4%), and suggesting prescription medications to physicians (52.0%). Respondent's age was negatively correlated with three functions related to pharmacotherapeutic regimens: designing regimens, monitoring effects of failed regimens, and monitoring outcomes. The most common negative experiences with pharmacists involved pharmacists' scaring the patient, dispensing unauthorized refills, and making inappropriate comments in the presence of patients. CONCLUSION: Future research with a larger, more representative sample of physicians will help explain this dynamic relationship. These preliminary results should be useful in training future physicians and pharmacists. PMID- 11029844 TI - Time and cost analysis of repacking medications in unit-of-use containers. AB - OBJECTIVES: To evaluate the effects of repacking drugs in unit-of-use containers in community pharmacies. The purpose of this study was to examine whether unit-of use repacking reduces routine mechanical "counting and pouring" to allow more time for pharmaceutical care. DESIGN: Simulation pilot study to evaluate the differences between the existing and proposed systems. Based on the literature, four variables--optimum pack size, time savings, packaging costs, and shelving requirements--were selected for evaluation. Historical prescription data from a chain were used in determining the sample drugs and their optimum pack sizes. Workflow analysis and time study were used to estimate the time savings. Manufacturer bottles, repack bottles, and shelves were measured to determine the impact of using unit-of-use containers on storage requirements. SETTING: Three community pharmacies in a major drugstore chain in Cincinnati, Ohio. RESULTS: The 25 fastest-moving solid oral dosage forms, representing 21.6% of all drugs dispensed by the chain, were selected as the sample drugs for the study. The workflow analysis and time study revealed that 0.79 minutes could be saved per prescription if repacked drugs were used. There was an increased cost of approximately $0.05 for every repack bottle used in place of a prescription vial. It was calculated that repacking in unit-of-use containers would increase storage requirements in the pharmacy by 2.5 times if current inventory levels were maintained. CONCLUSION: Repacking drugs in unit-of-use containers is potentially an inexpensive method to increase usable time in the pharmacy that does not require an increase in personnel or the purchase of additional technology at the store level. PMID- 11029845 TI - HMG-CoA reductase inhibitors: assessing differences in drug interactions and safety profiles. AB - OBJECTIVE: To review the cytochrome P450 system and associated metabolic differences between the HMG-CoA reductase inhibitors. DATA SOURCES: A MEDLINE search (1993-99) was conducted for English-language articles using key search terms including adverse drug reactions, cytochrome P450, drug metabolism, drug interactions, hydroxymethylglutaryl-CoA (HMG-CoA) reductase inhibitors, myopathy, and rhabdomyolysis. STUDY SELECTION: Review articles, clinical trials, and case reports concerning HMG-CoA reductase inhibitor metabolism, drug interactions, and adverse drug reactions were evaluated. DATA EXTRACTION: By the author. No software or assistants were used to extract information from the chosen studies. DATA SYNTHESIS: The cytochrome P450 enzymes, which can be divided into families, subfamilies, and isoenzymes, act as a major catalyst for drug oxidation in the liver. CYP3A4 is a major enzyme, accounting for about 60% of drug metabolic capacity in the liver and 70% of such function in the intestine. Lovastatin, simvastatin, and atorvastatin are substrates of CYP3A4, whereas fluvastatin is metabolized by CYP2C9. Pravastatin is not extensively metabolized by either of these isoenzymes; rather, it is transported into hepatocytes by a sodium independent, carrier-mediated uptake system that normally transports bile acids. Compared with other statins, pravastatin thus has a reduced potential for drug interactions with other substrates, inhibitors, or inducers of the CYP3A4 and CYP2C9 systems. CONCLUSION: Pharmacists must understand the functions of these enzymes to identify potential drug interactions, especially in high-risk patient populations, and to make appropriate therapeutic recommendations that prevent or minimize adverse clinical outcomes. PMID- 11029846 TI - Endometriosis: an overview of the disease and its treatment. AB - OBJECTIVE: To review endometriosis, its etiology, clinical presentation, and current management options. DATA SOURCES: Published articles identified through MEDLINE (1966-2000) using the search term "endometriosis" and the additional terms "etiology" and "treatment." Additional articles were identified from the bibliographies of the retrieved articles. DATA SYNTHESIS: Endometriosis, a disease that affects the physical health and emotional well-being of many women of reproductive age, is defined as the presence of endometrial tissue outside its normal location in the uterus. The disease ranges in severity from mild to severe, and patients may be asymptomatic or experience severe and potentially incapacitating symptoms, such as dysmenorrhea, dyspareunia, and infertility. The diagnosis can be confirmed only by direct visualization using laparoscopy and biopsy. The risk of endometriosis is increased in women who have an affected first-degree relative or who have shorter menstrual cycle lengths, longer duration of menstrual flow, and low parity. The etiology of endometriosis is not yet fully understand, but may involve retrograde menstruation, hereditary factors, and impaired immune function. Treatment should be individualized for each patient, taking into account the therapeutic goals, the extent of disease, symptomatology, and the woman's age and overall health. Treatment options include expectant management, hormonal therapies to suppress ovarian steroidogenesis and induce endometrial atrophy, and surgery to remove visible lesions or, as a last resort, the uterus and ovaries. CONCLUSION: Although the precise etiology of endometriosis remains a mystery, treatment options have improved considerably in recent years. Pharmacists are well positioned to identify women with unexplained pelvic pain or infertility that may be indicative of endometriosis and refer them to their physicians for further evaluation. Pharmacists also can play an important role in counseling patients about the safe and effective use of the various treatments for this disease and strategies to recognize and reduce adverse effects. PMID- 11029847 TI - Case study using descriptive analysis to estimate hidden costs in processing third party prescriptions. AB - OBJECTIVE: To identify and quantify additional hidden costs associated with processing third party prescriptions. DESIGN/SETTING: Using time and motion techniques, the frequency of rejected third party prescriptions and the time involved to resolve those rejections were measured in one supermarket chain pharmacy and one independent pharmacy. From this information, additional costs attributed to processing third party prescriptions were calculated. RESULTS: In a market where almost 95% of all prescriptions are third party reimbursed, payers rejected 18.7% and 22.3% of the prescriptions submitted by the supermarket chain and independent pharmacies, respectively. Additional or hidden costs for rejected prescriptions averaged $1.10 at the supermarket chain pharmacy and $1.54 at the independent pharmacy, with the difference attributable to the higher level of pharmacy staff involvement in resolving rejections at the independent pharmacy. When additional costs for all third party prescriptions were calculated, the average additional cost per third party prescription dropped to $0.44 for the supermarket chain pharmacy and $0.61 for the independent pharmacy. CONCLUSION: Increasing pharmacist availability for pharmaceutical care requires decreasing time spent in the dispensing process, especially resolving third party problems. Systems analysis and time and motion techniques were effectively used to more accurately measure the time and costs associated with processing third party prescriptions. Actual costs found in this study were considerable, but significantly less than those reported previously in studies using estimates and surveys. PMID- 11029848 TI - Third party pharmacy audits: can they be improved? AB - OBJECTIVE: To discuss issues surrounding the current auditing practices associated with third party insurance prescription claim programs. DATA SOURCES: Audit examples obtained during the last 10 years associated with third party audit methodology. SUMMARY: Many unattended issues are associated with third party pharmacy audits. Some issues for discussion are presented in an attempt to produce some possible solutions for the audit inequities. CONCLUSION: National professional organizations, insurance companies, and payers need to convene and formulate fair auditing guidelines. PMID- 11029849 TI - Falsely elevated systolic blood pressure with patient-operated blood pressure measuring devices. PMID- 11029850 TI - Pharmacy connect--a free drug program for community-dwelling elders. PMID- 11029851 TI - Cutting the obesity epidemic down to size. PMID- 11029852 TI - Please wait, so grandma gets vaccinated first. PMID- 11029853 TI - New biotech products for treatment/monitoring of diabetes. PMID- 11029854 TI - Physicians and pharmacists as fine artists. PMID- 11029855 TI - Advice from a walrus. PMID- 11029856 TI - 2000 Remington Lecture. A brief history of the present--as told by accreditation standards for pharmaceutical education. PMID- 11029857 TI - 2000 Prescott Lecture. Will pharmacy survive or will it prevail? A complex systems approach. PMID- 11029858 TI - Benefits and challenges of hormone replacement therapy. AB - HRT provides both prevention and treatment of osteoporosis. HRT is most likely beneficial in primary prevention of heart disease, and may have a role in preventing Alzheimer's disease and colon cancer. Prospective trials, such as the Women's Health Initiative, are needed to substantiate both benefits and risks. Decisions regarding HRT, as well as nonhormonal regimens, should be individualized. PMID- 11029859 TI - Building a year-round immunization program. AB - Each year, an estimated 55,000 to 80,000 people die from vaccine-preventable diseases. More pharmacists are needed to advocate, facilitate, and deliver immunizations throughout the year. Pharmacists who want to develop a year-round immunization program should begin by developing a comprehensive business plan. By implementing a successful year-round immunization program, pharmacists can reduce morbidity and mortality from a range of vaccine-preventable infections. PMID- 11029860 TI - Dietary supplements in women: responsible strategies for use. AB - The Dietary Supplement Health and Education Act of 1994 defined dietary supplements and helped to fuel the use of these products in the United States. Supplement users tend to be older, white, and female, and believe that diet affects health. Dietary supplements cannot replace a healthy diet, although they can be a valuable adjunct. Some supplements, notably iron and vitamin A, can cause toxicity in excess doses. Nutrient supplements that are particularly important for women include folic acid (during the childbearing years), calcium, and vitamin B12 (after age 50 years). PMID- 11029861 TI - Extraordinary infections: a focus on bioterrorism. AB - The possibility of biological or chemical terrorist attacks presents a very real threat. Current preparedness efforts are ongoing at federal, state, and local levels. A National Pharmaceutical Stockpile containing antibiotics, antidotes, and other pharmaceutical supplies has been established and continues to be developed. To provide enhanced local response capabilities, Metropolitan Medical Response Systems are being formed. Pharmacists are needed by their communities to become involved with preparedness efforts. PMID- 11029862 TI - Integrating herbs and botanicals into patient care. AB - Consumer use of herb and botanical preparations is burgeoning. Many common herbal remedies have proven efficacy, reported mainly in the European scientific literature. Although not mandated by law, responsible manufacturers of herbal products are working to establish efficacy, safety, and good manufacturing standards. Pharmacists should be alert for possible drug interactions between phytomedicines and pharmaceuticals, as well as potential adverse effects. Well informed pharmacists can provide useful counseling and education about herbs and botanicals. PMID- 11029863 TI - Lower extremity management in patients with diabetes. AB - Foot ulcers are a major cause of disability, morbidity, and mortality in people with diabetes. The three major etiological factors for diabetic foot ulcers are peripheral neuropathy, peripheral vascular disease, and infection. Tight control of blood glucose and proper foot care are essential for prevention of foot ulcers and lower extremity amputations. Treatment of lower extremity diabetic ulcers typically includes surgical debridement of the wound and appropriate oral or intravenous antibiotics. Becaplermin gel, a prescription, recombinant wound healing therapy, may be used to promote healing of diabetic foot ulcers in select patients. Pharmacists can make a significant contribution to the prevention of diabetic foot ulcers and lower extremity amputations. PMID- 11029864 TI - Obesity: managing comorbidities of the disease. AB - Obesity poses a serious health risk and is associated with a wide range of comorbid conditions. Obese patients with comorbid conditions such as type 2 diabetes and hypertension should be educated about the potential benefits of weight loss therapy. Pharmacists can offer consultative weight loss programs either on their own or by forming multidisciplinary teams with other practitioners, such as dietitians. PMID- 11029865 TI - Preventing dangerous drug interactions. AB - Although most drug interactions are clinically insignificant, some pose a significant risk. A basic knowledge of the mechanisms of drug interactions can help pharmacists to identify and avert potentially risky combinations. Review all medications, including dietary supplements and nonprescription drugs, when taking a medication history. Pay special attention to patients who take several medications, use herbal products, or use prescription medications associated with serious adverse events or toxicities. PMID- 11029866 TI - Sleep disorders: assisting patients to a good night's sleep. AB - Sleep is an active process consisting of macro- and microarchitectural rhythms, chronobiology, and neurochemical changes. Disruptions in sleep chronobiology, which may occur in shift workers and the elderly or due to jet lag, often respond to exogenous melatonin. Insomnia requires careful evaluation to determine contributions of comorbid medical conditions and drug effects. Treatment options include sleep hygiene principles and sedative hypnotic therapy. Zaleplon and zolpidem, short-acting benzodiazepines selective for omega-1 GABA receptors, are effective first-line agents for initiating sleep. PMID- 11029867 TI - Solid organ transplant: medication management issues. AB - Solid organ transplant patients are surviving longer today with a better quality of life. Although the incidence of acute rejection has decreased sharply in the past decade, chronic rejection remains an important problem. Goals for immunosuppression should focus equally on prevention of rejection and post transplant complications. Newer and more specific immunosuppressive agents are available that can improve patient outcomes with fewer adverse effects. Pharmacists can help to prevent post-transplant complications and improve outcomes with immunosuppressive therapy. PMID- 11029868 TI - Supportive care for the patient with Alzheimer's disease. AB - The prevalence of AD, the most common form of dementia, is likely to double in the next 20 years. Although the cause of AD is unknown, risk factors include aging, genetic predisposition, and head trauma. The cholinesterase inhibitors (tacrine, donepezil, and rivastigmine) are the only FDA-approved class of medications for AD. Behavioral problems associated with AD are a major reason for admission to long-term care facilities and a common cause of caregiver "burnout" or stress. PMID- 11029869 TI - Therapeutic options for GERD: what's the correct approach? AB - GERD occurs as a clinical spectrum of symptoms and complications. The primary goal of GERD therapy is to lessen esophageal exposure to acid, thus relieving symptoms, promoting mucosal healing, and preventing relapse and complications. Because GERD tends to recur after therapy is stopped, patients with healed erosive esophagitis usually require long-term maintenance therapy. Pharmacists have an important role in advising patients about therapeutic options for GERD, including lifestyle modifications and the proper use of nonprescription and prescription medications. PMID- 11029870 TI - Career awareness roundtable. AB - Employment prospects are excellent for recently graduated pharmacists. New pharmacy practitioners are employing their clinical, scientific, and business skills in an expanding array of fields. Starting salaries for newly graduated pharmacists are rising, although base salaries vary depending on the employer, field of practice, and area of the country. PMID- 11029871 TI - Liability for errors in expanded pharmacy practice: issues for technicians and pharmacists. AB - Industry trends, such as the growth of managed care and pharmacist-led cognitive services, have led to greater reliance on pharmacy technicians. In a recent USP study of medication errors, the dispensing function was the function in which technician errors were most likely to occur. Pharmacists and technicians must work as a team to reduce medication errors and improve quality assurance. Medication errors provide an opportunity to identify system problems, generate solutions, and implement corrective measures. PMID- 11029872 TI - Patient assessment tools: utilizing diagnostic and monitoring devices. AB - Pharmacists are becoming increasingly involved in the detection and monitoring of chronic disease states. A growing number of chronic diseases can be screened and monitored by pharmacists, including diabetes, dyslipidemias, hypertension, and osteoporosis. Devices for screening and monitoring are becoming less complex and more convenient, allowing for greater use in pharmacies and, in many cases, for patients use at home. The provision of pharmacist-led assessment services is becoming more widely accepted by patients, physicians, and some third-party payers. PMID- 11029873 TI - Regulatory issues in pain management. AB - Undertreatment of pain continues to be a serious public health problem. The use of opioid analgesics is considered essential for chronic pain management and palliative care. Pharmacists are encouraged to become involved in initiatives to improve pain management. PMID- 11029874 TI - Policy decisions about the human genome. AB - The application of genomic technology to drug development and therapy will lead to safer and more effective therapies. Elucidation of the human genome poses numerous social, ethical and legal issues, such as the possible misuse of genetic information to discriminate against a person in the workplace or through denial of insurance coverage. Pharmacists need to stay current with advances in genetic medicine and prepare themselves to use this information to optimize drug discovery and drug therapy. PMID- 11029875 TI - International nuclear pharmacy. PMID- 11029876 TI - Positron emission tomography: from production and distribution to drug research and clinical applications. AB - The radiopharmaceutical FDG has had a major impact on PET imaging in clinical medicine, particularly in the detection and staging of certain cancers. PET isotopes offer unparalleled insight into in vivo distribution of labeled drugs. PET imaging of blood flow, metabolism, and neuroreceptor characteristics may provide new perspectives on the mechanism of action and effects of drugs. PMID- 11029877 TI - Nuclear pharmacy: yesterday reviewed today. PMID- 11029878 TI - Reaction mechanism of bovine heart cytochrome oxidase. PMID- 11029879 TI - EGF receptor activation of the human gastrin gene: a tale of two zinc finger transcription factor families. AB - Gastrin is one of the oldest and most actively studied of the gastrointestinal peptides. Adult expression of the mature peptide is restricted to the stomach and duodenum. However, it is antral gastrin that is subject to feedback regulation from the counter-regulatory peptide somatostatin as a function of gastric pH. Yet little is known about the molecular steps required to modulate the synthesis of this peptide. Using cell culture models, a GC-rich regulatory element called gERE has been identified that binds two families of zinc finger transcription factors- Sp and ZBP. Competitive binding of these two transcription factor families in association with changes in their phosphorylation state appears to mediate the positive and negative activation of this gene in response to EGF receptor ligands. These findings contribute to our understanding of gastrin gene expression in the presence of the activated ras oncogene, e.g., during colonic transformation. PMID- 11029880 TI - A high level of physical fitness during thirties is a negative risk factor for colonic polyps during fifties. AB - This case-control study examined the relationship between the occurrence of colonic polyps in men in their fifties and the level of their physical fitness while in their thirties. The subjects consisted of 51 male Japan Self-Defense Forces officials in their fifties who had colonic polyps, as diagnosed by colonoscopic examination, and 46 control individuals. As an indicator of physical fitness between 30 and 39, we selected the best time recorded for each individual during that decade of life for the 1,500 meter Physical Fitness Test run. We calculated the odds ratio for polyps according to selected risk factors (including physical fitness), and a logistic regression analysis was used to adjust for possible confounding variables. Odds ratio (95% confidence interval, p value) for colonic polyps with physical fitness in the thirties was 0.36 (0.16 0.82, p < 0.05). With adjustment for the subjects' maximum Body Mass Index in both their thirties and fifties, and serum total cholesterol, the odds ratio was 0.39 (0.15-0.99, p < 0.05). We suggest that the occurrence of colonic polyps in men in their fifties can be reduced by maintaining a high level of physical fitness while in their thirties. PMID- 11029881 TI - A 35-year-old man with cerebral hemorrhage and pheochromocytoma: the second brain dead organ donor in Japan. AB - A 35-year-old man was brought into the emergency room of Keio University Hospital by ambulance because of a sudden onset of coma. His Glasgow Coma Scale was 3 and his blood pressure 150/100 mmHg. CT scanning revealed a subcortical hemorrhage 8 cm in diameter. His respiration deteriorated rapidly, and an emergency craniotomy was performed for hematoma removal and cerebral decompression. Postoperatively the patient remained in a deep coma (GCS = 3) requiring respiratory support. The family presented an organ donor card previously signed by the patient, and brain death was confirmed in accordance with Japan's transplant law. As a result of two tests conducted six hours apart brain death was confirmed on the 5th postoperative day. With the family's consent, the donor's heart, kidneys and skin were removed for organ transplantation to be performed in other institutions. An autopsy was performed after the removal of the organs and skin. An extensive subgaleal hemorrhage was found in the left cerebral hemisphere, and microscopic examination revealed extensive necrosis with karyolysis of neuronal cells, but no viable neuronal cells were found in the cerebrum. The brain stem was marked by edema, hemorrhage, infarction necrosis and neuronal cell loss. The cerebellum was swollen and congested and showed autolysis of the granular layer. These findings suggested brain death syndrome with respirator brain. Other autopsy findings included a huge pheochromocytoma in the right adrenal gland, bilateral bronchopneumonia, liver congestion and fatty metamorphosis with four cavernous hemangiomas, and mild chronic lymphocytic thyroiditis. This patient was the second brain-dead organ donor and the first brain-dead patient to undergo postmortem examination in Japan. PMID- 11029882 TI - [Cancers of the sweat glands: problems and controversies at the end of the 20th century]. PMID- 11029883 TI - [Detection and genetic typing of human papillomaviruses in cervical biopsies in Cagliari Province]. AB - INTRODUCTION: Human Papillomavirus (HPV) infection is considered an important risk factor for the development of cervical carcinoma. The aim of this work was to detect and genotype HPV DNA in cervical lesions from our Province. METHODS: HPV DNA was amplified by polymerase chain reaction (PCR) and genotyped by restriction fragment length polymorphism (RFLP) analysis. A total of 101 biopsies (43 koilocytic atypias, 20 CIN1, 19 CIN2, 17 CIN3 and 2 squamous carcinomas) were analyzed. RESULTS: HPV DNA was found in 41.8% of koilocytic atypias, in 95.0% of CIN1 and 100% of CIN2 and higher grade lesions. Only high risk genotypes were found in CIN2-3 and invasive carcinomas. HPV 16 was the most prevalent type in both CIN1 and CIN2-3 and the only HPV type found in situ and invasive carcinomas. HPV type 51 was found in 21.0% of CIN1 but it was rare in CIN2 and absent in more advanced lesions. PMID- 11029884 TI - [Histological picture of liver disease in thalassemia intermedia]. AB - Thalassemia Intermedia (TI) is a clinical definition in use for a spectrum of clinical conditions ranging in severity from the asymptomatic carrier status to the transfusion-dependent status. The histological lesions of the liver in patients affected by TI has not seen well characterized yet. The aim of this study was to define the histological picture of liver disease in TI. To this end we studied our pool of 22 Thalassemia Intermedia patients who underwent liver biopsy; none of them had blood transfusion. We took into consideration fibrosis according to Ishak's staging and iron overload according to Sciot's grading. Moreover, we determined the hepatic iron content by atomic absorption spectroscopy. Our results have shown that, in all patients studied, the histological picture was characterized by the increase in liver iron stores. By Perls's stain haemosiderin appeared to be stored mainly in hepatocytes in the majority of cases. Inspite of the large amounts of liver iron in these patients, fibrosis was absent or mild. We also noted the presence of iron-free-foci, in part of the patients. Our data show that the histological and histochemical picture of liver disease in Thalassemia Intermedia is very similar to that found in Thalassemia Major. The finding of iron-free-foci could induce to consider a follow up of these patients, even if asymptomatic. PMID- 11029885 TI - [Expression of p53 in gallbladder carcinoma and in dysplastic and metaplastic lesions of the surrounding mucosa]. AB - We studied the expression of p53 in 80 gallbladder carcinomas, 43 peritumoral mucosae, 5 adenomas and 20 mucosae of non tumoral gallbladders. Gallbladder cancers were classified according to WHO criteria. We found p53 overexpression in 51 out of the 80 gallbladder cancers (64%). p53 expression was variable in different histologic subtypes: 100% of intestinal type, 66% of papillary type, 83% of adenosquamous carcinomas and 66% of giant cells cancers showed immunoreactive cells. In well and moderately differentiated conventional gallbladder adenocarcinomas we found 60% of positive cases, while, among poorly differentiated conventional cancers, 83% were immunoreactive. All mucinous adenocarcinomas were p53-negative. In peritumoral dysplastic mucosae, p53 was expressed in 23 out 38 cases (60%), 22 (96%) of which were associated to a p53 positive adenocarcinoma. On the contrary, only 5 of the 15 p53-negative dysplastic lesions (33%) were associated to an p53-immunoreactive adenocarcinoma. Metaplastic lesions, of gastric and intestinal type, and adenomas were completely p53-negative. In conclusion, our data suggest that p53 expression is an early event, frequently involved in gallbladder carcinogenesis, and related to different histologic subtypes of gallbladder adenocarcinomas. PMID- 11029887 TI - [Demonstration of the calcified osseous component in decalcified bone marrow biopsies. Study of hematological cases]. AB - In order to investigate the possibility of the evidentiation of the mineralized component of decalcified bone a series of Jamshidi-type consecutive bone marrow biopsies for various hematological disorders were pre-stained with von Kossa modified procedure for calcium. Methodologically the controls showed reliable silver staining as far as localization (osteo-medullary interface and mineralization front) and preservation (after decalcification) were concerned. The results were: 1) a good morphology of bone marrow tissue also concerning immunohistochemical stainings; 2) a better overview of the osseous components and related artifacts as induced by biopsy, processing and sampling; 3) precise identification of the osteoid seams and remarkably of the relative angle of sectioning for an appropriate measurement; 4) better evidentiation of the remodelling osteoblastic-osteoclastic units; 5) visualization of the osteocytic lacunae and canaliculi and the mineralized matrix to some extent depending on their effective permeability. Summarizing the osteologic features were: normality or minimal abnormality of difficult interpretation; classical osteometabolic alterations; lesions specifically due to hematological disorders; various combination of these findings. Theoretical and practical aspects are discussed. In conclusion this methodological variant in comparison with the usual paraffin procedure clearly gives more information concerning osteometabolic evaluation in routine hematological biopsies; offers a vicarious or complementary approach to osteometabolic diseases; represents a conceptual stimulus to interpret diagnostically and prognostically the osseous pathology as determined by routinely encountered hematological disorders. PMID- 11029886 TI - Analysis of the spectrum of malformations in human fetuses of the second and third trimester of pregnancy with human triploidy. AB - Triploidy is constituted by an extra haploid set of chromosomes for a total of 69 chromosomes in humans. A "parent-of-origin" effect has been demonstrated by analysis of cytogenetic polymorphisms of triploidy pregnancies. Two distinct phenotypes of human triploid fetuses have been recognized according to the parental origin of the extra haploid set. The first one or triploidy of diandric type occurs when the extra haploid set of chromosomes arises from the father, the second one or triploidy of digynic type occurs when the extra haploid set of chromosomes arises from the mother. Diandric fetuses appear relatively well grown with a large placenta, while digynic fetuses show intrauterine growth retardation with a small placenta. Autopsy archive data files (1982-1998) of the Institute of Pathology, University of Heidelberg (Germany) were examined for fetuses with triploidy. We found 12 well-studied triploid fetuses (gestational age: 20 to 32 weeks). Eleven fetuses corresponded to the digynic type of triploidy and one fetus corresponded to the diandric type of triploidy. The spectrum of external malformations included a dysmorphic face (broad root of the nose, exophthalmos, low-set ears, micro-/retrognathia, microgenia, median cleft lip and/or palate, gnathoschisis, macroglossia), encephalocele, spina bifida, syndactyly, club or rocker-bottom feet, pes equino-valgus. More common internal malformations included ventricular septum defect of the heart, abnormal lobation of the lungs, and renal disease (agenesis, cysts). PMID- 11029888 TI - [Metastasis of pleural malignant mesothelioma to the brain and upper maxilla: description of 2 cases]. AB - Two cases of malignant mesothelioma metastatic to the brain and maxillary bone respectively are reported. This latter localization has never been described in literature. The diagnosis resided primarily on immunohistochemistry. PMID- 11029889 TI - [Blue nevus of the uterine cervix: description of a case and review of the literature]. AB - Blue nevus is a pigmented lesion of dermal melanocytes; the extracutaneous locations are uncommon. We report a case of a blue nevus of the uterine cervix in a 53 years old woman, with histochemical and immunohistochemical investigations. PMID- 11029890 TI - Epidermoid cyst of the ovary: a case report. AB - In the ovary, epidermoid cysts are rare and their origin is controversial. This rare lesion was uniformly an incidental finding in the study of hysterectomy specimens. The patient was a 48-year-old woman. Because of multiple leiomyomas she had a total hysterectomy and bilateral salpingo-oopherectomy operation. A Epidermoid cyst was found incidentally in the right ovary on the microscopic examination. PMID- 11029891 TI - [Viral dynamics in the course of HIV-1 infection: pathogenetic features and new therapeutic prospects]. PMID- 11029892 TI - [The autopsy and its merits: additional considerations]. PMID- 11029893 TI - [Trees with blood-colored wood]. PMID- 11029894 TI - Writing social work. PMID- 11029895 TI - Hidden voices. AB - Despite the rich history of care that has characterized the profession, social work has not been able to convey adequately its knowledge of the modest yet complex tasks involved in its role of social caretaking. The dominant voice of the formal culture, particularly in its emphasis on rationality and logic, does not create sufficient space or legitimacy for the experience of domestic and social caretaking to be conveyed. From personal and professional perspectives, this essay presents the vocabulary of care as the first voice of women and of social work and explores it as an avenue to better justify, dignify, and celebrate the humble but vital tasks carried out in all venues of social work practice. PMID- 11029896 TI - Reflections on empowerment-based practice. AB - Social work education presents students with theories for understanding human beings and the social-historical contexts they share. These assumptions are supposed to make social workers' roles, practices, and agencies make sense. But where do they come from? How are the theories selected--and from among what alternatives? The people or licensing bodies choosing or approving these options control knowledge and inhibit knowing beyond the parameters of what they have designated for social work's knowledge base. What were the knowledge requirements or factors used to select prevailing theories about human behavior and the social environment or about social work practice? What happens to practitioners when feelings about their work contradicts what they were taught they should be thinking? This article is a reflection of formal social work learning interacting with the author's own social work experience. The article discusses what receiving knowledge (as opposed to sharing in producing or generating it) means for a practice supposedly committed to the social work values of human dignity and social justice. The experiences that led to the author's work on empowerment based practice are presented and discussed. These experiences precipitated critical questioning that continues today. PMID- 11029897 TI - Lessons learned about working with men: a prison memoir. AB - From the very beginning of my career as a social worker, I have believed that the fullest and most effective use of self requires an integration of personal, professional, and spiritual-political values and beliefs into a framework that respects their dialectical tensions while striving for wholeness. In this very personal memoir, I tell of such a period in my life. The setting is a maximum security prison. I learned about mutuality, transparence, risk taking, openness, and vulnerability from men in the prison: inmates, the warden, and other prison staff. The lessons I learned have served as ground and guide for my subsequent psychotherapy practice with men, my own relational growth, and my commitment to the struggle for gender equality and social justice. PMID- 11029898 TI - Riding the mental health pendulum: mixed messages in the era of neurobiology and self-help movements. AB - Whereas professional disagreement about optimum mental health treatment is commonly understood, recognized, and discussed, the influence of opposing philosophies on patients is less often addressed. This article examines the long term distress created for the author, battling severe, recurrent depression, as she received advice from practitioners who offered widely varying treatment theories as the basis for differing recommendations. Medical knowledge based on advancing research in neurobiology has led to greater understanding of brain chemistry, more reliance on the use of pharmaceuticals, and an emphasis on other "physical" approaches to mental illness. At the same time, growing "self-help" recovery initiatives, with underlying theories analogous to cognitive therapy, continue a focus on emotional and behavioral self-direction. These messages can appear dramatically opposed in terms of expectations on the patient, when presented from separate perspectives that are not described in a context of the whole. The resulting confusion is an unfair imposition of the personal differences of philosophy among practitioners on patients in crisis seeking help. PMID- 11029899 TI - "Being" native versus "going native": conducting social work research as an insider. AB - The increasing cultural diversity among professional social workers has resulted in the need to examine critically some of the earlier notions about the epistemology, ontology, and methodology of social work research and practice. One outcome of these analyses about how and by whom research projects are carried out is the emergence of "native," "indigenous," or "insider" research in which scholars conduct studies with populations and communities and identity groups of which they are also members. This article reports the work of a native social work researcher who conducted an ethnographic study with her social identity group. The complex and inherent challenges of being both an insider with intimate knowledge of one's study population and an outsider as researcher are explored. Implications for social work research and practice with regard to native social work perspectives and methods also are discussed. PMID- 11029900 TI - Confronting stigma within the services system. PMID- 11029901 TI - What is the role of hypertechnology in social work today? PMID- 11029902 TI - Say it ain't so, Norm! Reflections on who we are. PMID- 11029903 TI - Responses to life after death with dignity: the Oregon experience. PMID- 11029904 TI - Responses to life after death with dignity: the Oregon experience. PMID- 11029905 TI - Responses to life after death with dignity: the Oregon experience. PMID- 11029906 TI - Responses to adoption policy in the United States: a word of caution. PMID- 11029907 TI - Responses to adoption policy in the United States: a word of caution. PMID- 11029908 TI - Model state social work practice act. PMID- 11029909 TI - Retired social workers. PMID- 11029910 TI - Choice and empowerment for battered women who stay. PMID- 11029911 TI - Sexual contact with clients. PMID- 11029912 TI - Bricks and clicks: whither our customers concerns? PMID- 11029913 TI - Workplace adjustment and intergenerational differences between matures, boomers, and xers. AB - The authors explored the factors influencing occupational adjustment related to workplace stress among 413 nurses at a Midwestern pediatric hospital. Among critical factors found in responses to their questionnaire and follow-up focus groups were differences in work adjustment and intergenerational conflicts. Both real and perceived workplace stress can manifest itself both fiscal and human costs by increasing turnover, absenteeism and worker's compensation claims as well as "faulty products and negative behaviors." Baby boomers (those born between 1946 and 1964) and Generation Xers (those born between 1965 and 1981) reported quite different issues and perceptions of occupational stress. PMID- 11029914 TI - Expanded role of nursing in ambulatory managed care. Part I: Literature, role development, and justification. AB - New expanded roles in nursing are developing in response to the health system switch from private, episodic care of individuals to health maintenance organizations (HMO's) focusing on disease prevention and promotion of health for populations of persons with chronic illness. This evaluation project explored the role of the nurse clinician in population and case management at a large HMO in the Northwest United States. In Part I of this two-part series, a literature review, is presented on chronic illness, ambulatory nursing, expanded role nursing, and nursing in HMOs. The development of the expanded role nurse clinician at the managed care organization is also described. Results could be used by HMOs to attract and retain nurses, improve care processes, and decrease costs in ambulatory care. PMID- 11029915 TI - The marketability of nurse practitioners in New York City. AB - Until recently nurse practitioners (NPs) in independent practices have almost exclusively served Medicaid populations, and, most NP care has been provided by NP employees in physicians' offices or HMOs. The author explores the willingness of affluent or insured private pay patients to choose NPs as their primary care providers by using both focus groups and telephone interviews of adults between 25 and 50 years of age. Findings included the common perception that physicians were best prepared by education at diagnosing illness and valued for their overall expertise and likelihood to treat patients with respect. Nurses were most liked for their "compassion and caring," their willingness to listen to client concerns, and focus on disease management and health promotion. Nurses were least liked for having "bad attitudes" and being overworked. Only 23% of the survey participants were familiar with the term "advanced practice nurse" while 76% had heard of NPs. PMID- 11029916 TI - Private pay clients in adult day services. AB - Data collected for all 3,992 participants in Maryland's adult day care programs as of December 31, 1993 showed that the majority of clients were supported by publicly supported "welfare" funding sources. The authors examined the differences in characteristics and service needs between the public payer population and private pay patients, including those who were charged on a sliding scale. They found that private pay clients were not dependent on others to "initiate service arrangements when they want and can find them." Private pay clients were twice as likely to be self or family referrals and more likely to be older, white, and living with a spouse or adult child. Private pay patients were also much more likely to have a history of the recent onset of cognitive impairment and associated memory loss but less likely to fall into the more ADL dependent categories. The nuclear families of private pay clients who are primary caregivers are described as the "critical marketing targets" and that services specifically for the cognitively impaired should be developed. PMID- 11029917 TI - TIQ (Technical IQ)--a survival skill for the new millennium. AB - Managing in the new technocracy of the new millennium will be challenging but also much more exciting than the work we do now. Twenty years ago, nurse managers weren't held accountable for the level of knowledge about financial outcomes that is expected now. The same will be true for technology. With electronic medical records, innovations in medical treatments, and the impact of the information and technologic revolution on the entire world, the interface between technology assessment, operationalizing technology, and continually upgrading staff to know what the implications of the technology revolution are will be imperative for success. That means that we must influence schools of nursing to include technology in their content now, and to help us prepare this generation of health care leaders to take a full seat at the leadership table to manage these complexities in the future. The success of the leader of the future will be measured by that person's ability to integrate the very complex issues of patient care and technology in a way that makes sense for patients, the organization, and the staff who will be working in a very complex environment. The leader's TIQ will be as important as other intelligences in the next 20 years. PMID- 11029918 TI - Integrated delivery systems: Mercy Health Services, Part I. Interview by SueEllen Pinkerton. PMID- 11029919 TI - Is unionization the answer for nurses and nursing? PMID- 11029920 TI - Nursing informatics: using uniform language in patient care documentation. PMID- 11029921 TI - Job sharing: an option for professional nurses. PMID- 11029922 TI - Answering tough questions on interview. PMID- 11029923 TI - HIPAA: the next frontier. PMID- 11029924 TI - Presidential address by Mrs. S.A. Samuel, president, TNAI, delivered at the inaugural function of the XVIII TNAI biennial (67th) conference, Imphal (Manipur) on October 28, 1999. PMID- 11029925 TI - Intranatal care by traditional birth attendant--a case study approach. PMID- 11029926 TI - Oncology nursing--expanded role. PMID- 11029927 TI - Schizophrenia patients: duration of hospital care and returning to the self care ability. PMID- 11029928 TI - Following the Nightingale legacy. PMID- 11029929 TI - Empowerment through learning: the University of Toronto IIWCC completes its first course. PMID- 11029930 TI - Mixed up about PPS case-mix in home care? PMID- 11029931 TI - Culture, health, and healing. AB - Americans are increasingly interested in and willing to participate in alternative or adjunctive healthcare, which is sometimes called complementary healthcare. Some believe this is because of wide-spread cultural diversity or an increasing awareness of options in health practice, while others contend it is a reaction to dissatisfaction with Western health practices. Diversity among cultures and subcultures has posed challenges for clinicians. This article addresses the impact of culture on health and healing, examining alternative health practices and their impact on chronic care, specifically chronic wound care. PMID- 11029932 TI - Outcomes of venous ulcer care: results of a longitudinal study. AB - A longitudinal study involving 81 patients with venous ulcers was conducted to explore the outcomes and cost of wound care in a home healthcare (HHC) setting and an outpatient care setting. Ulcers were managed with a saline gauze or hydrocolloid dressing and compression hosiery, or covered with an Unna's boot. Outcomes did not vary between physician's office and home care. Patients preferred home care, but costs and charges were much higher for HHC than for patients managed in the physician's office. Recurrence rates and costs varied greatly. Eighty-eight percent of ulcers in the saline dressing group did not heal or recurred compared to 21% of ulcers in the Unna's boot and 13% of ulcers in the hydrocolloid dressing group. The data also suggest hydrocolloid dressings are more cost-effective than Unna's boot or saline-gauze dressings. Controlled clinical studies to ascertain the cost-effectiveness of venous ulcer care in different patient care settings and the use of different treatment modalities, as well as care system oriented toward outcome for the patient rather than service, design, and distribution, are needed. PMID- 11029933 TI - Prevention of pressure ulcers in acute and long-term care facilities in Finland: results of a survey. AB - The purpose of this study was to identify those healthcare organizations that have a high incidence of pressure ulcers, and to determine what their staffs do to prevent pressure ulcer formation. The sample was formed from 11 hospitals in one large city on a certain day in Finland in 1998. The researchers sent questionnaires to 154 hospital units and achieved a 94% response rate. Psychiatric; gynecologic; obstetric; and eye, ear, nose, and throat units were not included. The data were collected using two questionnaires: The first gathered data about the organization and the second about the patients. Fifty seven percent of the units surveyed reported having patients with pressure ulcers. Of these units, 45% were acute and 55% were long-term care. Thirty-nine percent of all units had a pressure ulcer team. Units with pressure ulcer patients had a staffing level of 0.6 registered nurses and practical nurses per bed, compared with 0.7 registered nurses and practical nurses per bed for those units without pressure ulcer patients. The average length of stay on the unit was less for those without pressure ulcers compared to those with ulcers (P < 0.001) and only 18% of the units without pressure ulcers had a pressure ulcer team. According to the results, those units with pressure ulcer patients identified the need for more preventive measures more frequently than the units without ulcer patients. In conclusion, pressure ulcers seem to predominate in long-term care settings, and the educational level of healthcare staffing seems to impact the occurrence of pressure ulcers. Common preventive measures are used by the staff in both acute and long-term care settings. PMID- 11029934 TI - Neonatal wound dehiscence and the subsequent healing process: a case study. AB - Many neonates require abdominal surgery for a variety of reasons, including necrotizing enterocolitis (NEC). Secondary complications of abdominal surgery include alterations in skin integrity and potential wound dehiscence. These alterations may actually worsen when treated with products "traditionally" used postoperatively. The author simultaneously utilized basic wound care products with currently recognized therapies in managing a 29-week premature infant who experienced dehiscence secondary to bowel repair. By utilizing the correct products and incorporating the principles of moist wound healing and occlusion, this Stage III/IV wound, measuring 12 cm x 3 cm, closed within 35 days of dehiscence. A team-oriented and coordinated approach proved that wounds can, and will, improve, even in this fragile population. PMID- 11029935 TI - Outcomes from the view in the bed. PMID- 11029936 TI - Interdisciplinary clinical education: evaluating outcomes of an evolving model. PMID- 11029937 TI - Home health care, outcomes management, and the Land of Oz. PMID- 11029938 TI - Using clinical models to frame outcomes evaluation: the Arizona Nurses' Association Nursing Report Card Project. AB - The American Nurses Association (ANA) inaugurated Nursing's Safety and Quality Initiative in 1994. This multiphased initiative provides a framework for the application of the Nursing Care Report Card for Acute Care that aims to evaluate the nature and strength of the linkages between nursing care and nursing sensitive quality indicators in the acute care setting. Arizona is one of six states participating in this investigation. This article describes the development of the Arizona Nurses' Association (AzNA) Nursing Report Card Project and associated model. PMID- 11029939 TI - Design and implementation of an outcomes management model. AB - In today's capitation-based reimbursement environment, acute care staff nurses must coordinate patient care considering the full continuum of care. To do so effectively, staff nurses need tools to accurately predict patient needs, adjust service intensity accordingly, and evaluate the outcomes of the care provided. The design and implementation of a model to support acute care staff nurses in that effort is described. The model's implementation was evaluated, in part, using a ten-item pre- and post-implementation survey. The survey showed that staff nurses who participated in the final educational offering on the model increased significantly their reported use of aggregate data for planning care. To achieve the greatest impact, the model needs to evolve so that it is applied earlier in the care process than at initial hospitalization. Ideally, this should occur during a wellness visit or enrollment in a health plan. PMID- 11029940 TI - Creating an outcomes framework. AB - Four constructs used to build a framework for outcomes management for a large midwestern tertiary hospital are described in this article. A system framework outlining a model of clinical integration and population management based in Steven Shortell's work is discussed. This framework includes key definitions of high-risk patients, target groups, populations and community. Roles for each level of population management and how they were implemented in the health care system are described. A point of service framework centered on seven dimensions of care is the next construct applied on each nursing unit. The third construct outlines the framework for role development. Three roles for nursing were created to implement strategies for target groups that are strategic disease categories; two of those roles are described in depth. The philosophy of nursing practice is centered on caring and existential advocacy. The final construct is the modification of the Dartmouth model as a common framework for outcomes. System applications of the scorecard and lessons learned in the 2-year process of implementation are shared PMID- 11029943 TI - Evaluation of fiscal and treatment outcomes in major joint replacement. AB - The market demand for major arthroplasty procedures is increasing. This descriptive comparative study was conducted to examine clinical and fiscal outcomes in total joint arthroplasty patients discharged either to home or to a subacute unit. The post-acute care setting was self-selected by patients after information was provided on both options. The Self-Administered Joint Rating Questionnaire served as the primary data collection tool. Age, health status, and living alone were significant factors in post-acute care site selection. Although there were no significant differences in clinical outcomes between the two groups, overall costs were substantially different. Opportunities to maintain outcome status, while reducing total costs, in the subacute group are discussed. PMID- 11029942 TI - Using NOC outcome of risk control in prevention, early detection, and control of hypertension. AB - Hypertension is a major health problem addressed by nurses in all settings. This article reviews long-term hypertension outcomes and asserts the need to use intermediate outcomes focusing on prevention and early detection to effect change in the prevalence of hypertension and its complications. The Nursing Classification Outcome of Risk Control is presented as a framework to evaluate the state of the science and to develop a research agenda related to hypertension "Risk Control." PMID- 11029941 TI - Stroke management: beginnings. AB - This article outlines the evolution of a stroke program at one large midwestern tertiary care hospital. Implementation of unit-based care coordination, a standard order set, a clinical pathway, a nurse case manager, and ongoing multidisciplinary review were some of the strategies used to demonstrate improvement in outcome measures. Improvements were documented, including computed tomography (CT) scans being performed more quickly when patients arrived at the Emergency Department, decreased costs, decreased readmission rates, and reduced length of stay. PMID- 11029944 TI - Have we lost the plot? PMID- 11029945 TI - Symphysis-fundal height measurement. PMID- 11029946 TI - Confidential Enquiry into Stillbirths and Deaths in Infancy (CESDI). Highlights of the 6th annual report. Part 4: Record keeping. PMID- 11029947 TI - Breastfeeding: early problems. PMID- 11029948 TI - The midwife 2010. Exploring the impact of genetics on the future of midwifery. PMID- 11029949 TI - Acting the part. Using drama to empower student midwives. PMID- 11029950 TI - Improving our working lives. PMID- 11029951 TI - Evidence-based midwifery care. Evaluation of a pilot action learning programme. PMID- 11029952 TI - Brief encounters. Picking up signals of relationship distress. PMID- 11029953 TI - The demise of a midwifery group practice ... something works well for mothers and midwives: let's axe it! PMID- 11029954 TI - Motivational interviewing. PMID- 11029955 TI - Breastfeeding confusion? PMID- 11029956 TI - A really minimum F. PMID- 11029957 TI - Pleased to meet you. Tricia Anderson. PMID- 11029958 TI - A surprising outcome. PMID- 11029959 TI - It works. It's fair. It's Medicare. PMID- 11029960 TI - Latex allergy: more than a rash on the hands. PMID- 11029961 TI - Nurse in profile. Dr Linda Shields. PMID- 11029962 TI - Human overfeeding experiments: potentials and limitations in obesity research. PMID- 11029963 TI - Low dose oral iodized oil for control of iodine deficiency in children. AB - In areas where iodized salt is not available, oral iodized oil is often used to correct I deficiency despite a lack of consensus on the optimal dose or duration of effect, particularly in children, a main target group. Annual doses ranging from 400 to 1000 mg have been advocated for school-age children. Because lower doses of iodized oil have been shown to be effective in treating I deficiency in adults, the aim of this study was to evaluate the efficacy and safety of a low dose of oral iodized oil in goitrous I-deficient children. Goitrous children (n 104, mean age 8.4 years, range 6-12 years, 47% female) received 0.4 ml oral iodized poppyseed-oil containing 200 mg I. Baseline measurements included I in spot urines (UI), serum thyroxine (T4), whole blood thyroid-stimulating hormone (TSH), and thyroid-gland volume using ultrasound. At 1, 5, 10, 15, 30 and 50 weeks post-intervention, UI, TSH and T4 were measured. At 10, 15, 30 and 50 weeks, thyroid-gland volume was remeasured. At 30 and 50 weeks the mean percentage change in thyroid volume from baseline was -35% and -41% respectively. The goitre rate fell to 38% at 30 weeks and 17% at 50 weeks. No child showed signs of I-induced hypo- or hyperthyroidism. UI remained significantly increased above baseline for the entire year (P < 0.001); the median UI at 50 weeks was 97 micrograms/l, at the World Health Organization cut-off value (100 micrograms/l) for I-deficiency disorders risk. In this group of goitrous children, an oral dose of 200 mg I as Lipiodol (Guerbert, Roissy CdG Cedex, France) was safe and effective for treating goitre and maintaining normal I status for at least 1 year. PMID- 11029964 TI - Cholesterol-lowering potential in human subjects of fat from pigs fed rapeseed oil. AB - The possibility of achieving blood-lipid-lowering characteristics of pig fat by increasing the content of unsaturated fat in pig feed was evaluated. Three pig feeding regimens were applied: basal feed (no added fat or vitamin E), basal feed + rapeseed oil (60 g/kg feed), and basal feed + rapeseed oil (60 g/kg) + vitamin E (200 mg/kg). Meat and meat products from the three pig groups were incorporated into diets providing 86 g pig fat/10 MJ. The diets were served to twelve healthy human male subjects for 3 weeks each in a randomised crossover design. The diets prepared from pigs fed rapeseed oil had a lower content of saturated fatty acids (approximately 9 v. 11% of energy) and a higher content of polyunsaturated fatty acids (approximately 6 v. 4% of energy) than the diet prepared from pigs fed the basal feed. Diets based on fat from pigs fed the rapeseed oil resulted in significantly lower (approximately 4%, P = 0.019) total serum cholesterol concentration compared with the diet from pigs fed the basal feed. No differences were observed in LDL-, HDL- or VLDL-cholesterol, or in triacylglycerol or VLDL triacylglycerol concentrations. Addition of vitamin E to the pig feed resulted in only a minor increase in vitamin E content in the human subjects' diet and the vitamin E content was low in all three pig diets. Plasma vitamin E concentration in the human subjects at the end of the period with diets from pigs fed rapeseed oil without vitamin E was significantly lower (P = 0.04) than in the other two diet periods. In conclusion, an increased content of rapeseed oil in pig feed changes the fatty acid composition of the pig fat in a way that has a potential to reduce blood cholesterol concentrations in human subjects. However, intake of pig fat with a higher content of unsaturated fatty acids needs to be matched by a higher dietary intake of vitamin E. PMID- 11029965 TI - Response of putative indices of copper status to copper supplementation in human subjects. AB - No sensitive functional index is currently available to assess Cu status in healthy human populations. This study evaluated the effect of Cu supplementation on putative indices of Cu status in twelve women and twelve men, aged between 22 and 45 years, who participated in a double-blind placebo controlled crossover study. The study consisted of three 6-week supplementation regimens of 3 mg CuSO4, 3 mg Cu-glycine chelate and 6 mg Cu-glycine chelate, each separated by placebo periods of equal length. Women had significantly higher caeruloplasmin oxidase activity (P < 0.001), caeruloplasmin protein concentration (P < 0.05), and serum diamine oxidase activity (P < 0.01) at baseline than men. Erythrocyte and leucocyte superoxide dismutase activity, leucocyte cytochrome c oxidase activity, and erythrocyte glutathione peroxidase activity did not respond to Cu supplementation. Platelet cytochrome c oxidase activity was significantly higher (P < 0.01), after supplementation with 6 mg Cu-glycine chelate in the total group and in women but did not change in men. Caeruloplasmin oxidase activity was significantly higher (P < 0.05), in men after supplementation with 3 mg Cu glycine chelate, while caeruloplasmin protein concentration was significantly lower in men after supplementation with 6 mg Cu-glycine chelate (P < 0.05). Serum diamine oxidase activity was significantly higher after all supplementation regimens in the total group and in both men and women (P < 0.01). These results indicate that serum diamine oxidase activity is sensitive to changes in dietary Cu intakes and may also have the potential to evaluate changes in Cu status in healthy adult human subjects. PMID- 11029966 TI - Effect of abomasal glucose infusion on alanine metabolism and urea production in sheep. AB - The effect of abomasal infusion of glucose (120 kJ/d per kg body weight (BW)0.75, 758 mmol/d) on urea production, plasma alanine-N flux rate and the conversion of alanine-N to urea was studied in sheep offered a low-N diet at limited energy intake (500 kJ/d per kg BW0.75), based on hay and grass pellets. Glucose provision reduced urinary N (P = 0.040) and urea (P = 0.009) elimination but this was offset by poorer N digestibility. Urea-N production was significantly reduced (822 v. 619 mmol/d, P = 0.024) by glucose while plasma alanine-N flux rate was elevated (295 v. 342 mmol/d, P = 0.011). The quantity of urea-N derived from alanine tended to be decreased by glucose (127 v. 95 mmol/d) but the fraction of urea production from alanine was unaltered (15%). Plasma urea and alanine concentrations (plus those of the branched chain amino acids) decreased in response to exogenous glucose, an effect probably related to enhanced anabolic usage of amino acids and lowered urea production. PMID- 11029967 TI - Supplementation with alpha-tocopherol or a combination of alpha-tocopherol and ascorbic acid protects the gastrointestinal tract of iron-deficient rats against iron-induced oxidative damage during iron repletion. AB - Recently we have shown the susceptibility of Fe-deficient rat intestine to oxidative damage during Fe repletion. The role of dietary antioxidants like ascorbic acid, alpha-tocopherol and a combination of both in counteracting the oxidative stress was tested in this study. Five groups of thirteen weanling WKY female rats were fed with an Fe-deficient diet for a period of 5 weeks. Another set of thirteen rats received an Fe-sufficient diet and served as the control group (Con). Oral administration of either vehicle (D), 8 mg Fe alone (D+) or in the presence of 24 mg ascorbic acid (D+ + C), 40 mg alpha-tocopherol (D+ + E) or a combination of both (D+ + C + E) per d for 15 d was carried out in Fe-depleted rats. The impact of this treatment protocol on Fe status, oxidative stress and antioxidant status at the site of Fe absorption was assessed. It was observed that though the indicators of Fe status were normalised on Fe supplementation, the oxidative stress as reflected by the levels of both thiobarbituric-acid reactive substances (TBARS) and protein carbonyls were significantly greater in D+ and D+ + C compared to D+ + E, D+ + C + E and Con groups. The mucosal cell DNA damage was seen in D+, D+ + C and D+ + E groups on electrophoresis. Functional integrity as assessed by the activities of alkaline phosphatase and lys-ala dipeptidyl aminopeptidase were normalized in all the groups treated with the antioxidant(s). There were significant positive alterations in some of the endogenous antiperoxidative systems and in serum caeruloplasmin activity in D+ + E and D+ + C + E groups. Paradoxically, serum ascorbate levels were significantly lower in D+ + C than in D+ + E and D+ + C + E groups. This could be due to the protection offered by alpha-tocopherol in the presence of Fe. It is concluded that supplementation of alpha-tocopherol alone or in combination with ascorbic acid protects the gastrointestinal tract of Fe-deficient rats against Fe-mediated oxidative damage during Fe repletion. However, ascorbic acid alone does not protect the gastrointestinal tract against Fe-induced oxidative stress. PMID- 11029968 TI - Dietary n-3 fatty acids affect mRNA level of brown adipose tissue uncoupling protein 1, and white adipose tissue leptin and glucose transporter 4 in the rat. AB - We examined the effect of dietary fats rich in n-3 polyunsaturated fatty acids (PUFA) on mRNA levels in white and brown adipose tissues in rats. Four groups of rats were fed on a low-fat diet (20 g safflower oil/kg) or a high-fat diet (200 g/kg) containing safflower oil, which is rich in n-6 PUFA (linoleic acid), or perilla (alpha-linolenic acid) or fish oil (eicosapentaenoic and docosahexaenoic acids), both of which are rich in n-3 PUFA, for 21 d. Energy intake was higher in rats fed on a high-safflower-oil diet than in those fed on low-fat or high-fish oil diet, but no other significant differences were detected among the groups. Perirenal white adipose tissue weight was higher and epididymal white adipose tissue weight tended to be higher in rats fed on a high-safflower-oil diet than in those fed on a low-fat diet. However, high-fat diets rich in n-3 PUFA, compared to a low-fat diet, did not increase the white adipose tissue mass. High fat diets relative to a low-fat diet increased brown adipose tissue uncoupling protein 1 mRNA level. The increases were greater with fats rich in n-3 PUFA than with n-6 PUFA. A high-safflower-oil diet, compared to a low-fat diet, doubled the leptin mRNA level in white adipose tissue. However, high-fat diets rich in n-3 PUFA failed to increase it. Compared to a low-fat diet, high-fat diets down regulated the glucose transporter 4 mRNA level in white adipose tissue. However, the decreases were attenuated with high-fat diets rich in n-3 PUFA. It is suggested that the alterations in gene expression in adipose tissue contribute to the physiological activities of n-3 PUFA in preventing body fat accumulation and in regulating glucose metabolism in rats. PMID- 11029969 TI - Postnatal regulation of myosin heavy chain isoform expression and metabolic enzyme activity by nutrition. AB - Development of muscle is critically dependent on several hormones which in turn are regulated by nutritional status. We therefore determined the impact of mild postnatal undernutrition on key markers of myofibre function: type I slow myosin heavy chain (MyHC) isoform, myosin ATPase, succinate dehydrogenase and alpha glycerophosphate dehydrogenase. In situ hybridization, immunocytochemistry and enzyme histochemistry were used to assess functionally distinct muscles from 6 week-old pigs which had been fed an optimal (6% (60 g food/kg body weight per d)) or low (2% (20 g food/kg per d)) intake for 3 weeks, and kept at 26 degrees C. Nutritional status had striking muscle-specific influences on contractile and metabolic properties of myofibres, and especially on myosin isoform expression. A low food intake upregulated slow MyHC mRNA and protein levels in rhomboideus by 53% (P < 0.01) and 18% (P < 0.05) respectively; effects in longissimus dorsi, soleus and diaphragm were not significant. The oxidative capacity of all muscles increased on the low intake, albeit to varying extents: longissimus dorsi (55%), rhomboideus (30%), soleus (21%), diaphragm (7%). Proportions of slow oxidative fibres increased at the expense of fast glycolytic fibres. These novel findings suggest a critical role for postnatal nutrition in regulating myosin gene expression and muscle phenotype. They have important implications for optimal development of human infants: on a low intake, energetic efficiency will increase and the integrated response to many metabolic and growth hormones will alter, since both are dependent on myofibre type. Mechanisms underlying these changes probably involve complex interactions between hormones acting as nutritional signals and differential effects on their cell membrane receptors or nuclear receptors. PMID- 11029970 TI - The effect of vitamin C or vitamin E supplementation on basal and H2O2-induced DNA damage in human lymphocytes. AB - There is a wealth of epidemiological information on antioxidants and their possible prevention of disease progression but very little of the research on antioxidants has involved intervention studies. In this study, the potential protective effect of vitamin C or E supplementation in vivo against endogenous and H2O2-induced DNA damage levels in lymphocytes was assessed. The supplementation involved fourteen healthy male and female non-smokers mean age 25 53 (SD 1.82) years, who were asked to supplement an otherwise unchanged diet with 1000 mg vitamin C daily for 42 d or 800 mg vitamin E daily for 42 d. DNA damage in H2O2-treated peripheral blood lymphocytes (PBL) and untreated PBL before and after supplementation, and during a 6-week washout period was assessed using an ELISA. At each sampling time-point, the red cell concentrate activities of superoxide dismutase, catalase and glutathione peroxidase were also determined. Supplementation with vitamin C or vitamin E decreased significantly H2O2-induced DNA damage in PBL, but had no effect on endogenous levels of DNA damage. The activities of the antioxidant enzymes superoxide dismutase and glutathione peroxidase were suppressed during the supplementation period. These supplementation regimens may be used to limit the possible adverse effects of reactive oxygen species (including those produced during the course of an immune response) on lymphocytes in vivo, and so help to maintain their functional capacity. PMID- 11029971 TI - Effect of postprandial lipaemia and Taq 1B polymorphism of the cholesteryl ester transfer protein (CETP) gene on CETP mass, activity, associated lipoproteins and plasma lipids. AB - A large number of studies in recent years have investigated the effects of hyperlipidaemias and diabetes on cholesteryl ester transfer protein (CETP) on neutral lipid transfer activity and plasma lipids. There has been an ongoing debate as to whether CETP is pro- or anti-atherogenic as it provides a mechanism for the transfer of cholesterol from the cardioprotective HDL subfraction to the potentially atherogenic LDL subfraction. This study was designed to investigate whether there was significant variability of CETP mass and activity in a large normolipidaemic population and whether there is an association between CETP and plasma lipoprotein composition. The presence of a known polymorphism of CETP gene (Taq 1B) was investigated to see if there was any association between this polymorphism and CETP mass and activity, and plasma lipids. There was significant (P < 0.0001) increase in CETP mass and activity in plasma postprandially at 6 h. Using multiple stepwise regression analysis there was significant association with fasting CETP mass and activity (beta = 0.055; P = 0.002) and triacylglycerol rich lipoprotein (beta = 0.013; P = 0.005) and postprandial CETP mass (beta = 0.254; P = 0.007). Repeated-measures analysis showed a strong association between the absence of Taq 1B polymorphism and low CETP mass and elevated HDL- and HDL2 cholesterol and HDL-phospholipid concentrations than did those who were homozygous or heterozygous for the presence of the restriction site. PMID- 11029972 TI - Effects of dietary sulfur-containing amino acids on oxidative damage in rat liver caused by N-nitrosodimethylamine administration. AB - Effects of dietary protein and S-containing amino acids on oxidative damage were investigated in rat liver. After feeding rats for 3 weeks from weaning, lower GSH levels and higher metallothionein (MT) levels were found in the liver of rats fed on a 10% soyabean-protein-isolate (SPI)-based diet than those fed on a 10% casein based diet. After injection of N-nitrosodimethylamine (NDMA) at 20 mg/kg body weight, increases in lipid peroxide, determined as thiobarbituric-acid reactive substances (TBARS), and gamma-glutamyltransferase (GGT) activity in the liver were observed in the 10% SPI diet group. By supplementing the 10% SPI diet with 0.3% cystine or methionine, GSH levels were increased, while MT levels were decreased, and elevation in TBARS levels after NDMA injection was diminished. On the other hand, increase in GGT activity could be prevented only by methionine supplement. On a 20% SPI or casein diet, TBARS concentration and GGT activity were not altered after NDMA injection with concomitant increase in GSH levels and decrease in MT levels. These results indicate that sufficient amounts of methionine and cystine in a diet are important to protect the liver from oxidative damage after NDMA administration, and GSH plays a primary role in the cellular protective function when compared with MT. PMID- 11029973 TI - No energy compensation at the meal following exercise in dietary restrained and unrestrained women. AB - The objective of this investigation was to compare the acute effects of exercise and diet manipulations on energy intake, between dietary restrained and unrestrained females. Comparisons of two studies using an identical 2 x 2 repeated-measures design (level of activity (rest or exercise) and lunch type (high-fat or low-fat)) including thirteen dietary unrestrained and twelve restrained females were performed. Energy expenditure during the rest session was estimated and the energy cost of exercise was measured by indirect calorimetry. Relative energy intake was calculated by subtracting the energy expenditure of the exercise session from the energy intake of the test meal. Post-meal hedonic ratings were completed after lunch. Energy intake and relative energy intake increased during high-fat conditions compared with the low-fat, independently of exercise (P < 0.001). There was a positive relationship between dietary restraint scores and energy intake or relative energy intake in the rest conditions only (r 0.54, P < 0.01). The decrease of relative energy intake between the rest and exercise conditions was higher in restrained than in unrestrained eaters (P < 0.01). These results confirm that a high-fat diet reversed the energy deficit due to exercise. There was no energy compensation in response to an acute bout of exercise during the following meal. In restrained eaters, exercise was more effective in creating an energy deficit than in unrestrained eaters. Exercise may help restrained eaters to maintain control over appetite. PMID- 11029974 TI - The acute effect of D-tagatose on food intake in human subjects. AB - A double-blind randomized crossover study was performed with nineteen normal weight men to investigate the effect on subsequent ad libitum food intake of replacing 29 g sucrose with 29 g D-tagatose as sweetener to a breakfast meal. D Tagatose is a malabsorbed stereoisomer of fructose with potential application as a bulk sweetener. Food intake was measured at lunch offered 4 h after the breakfast meal, during the afternoon with access to abundant snacks, and finally at a supper buffet 9 h after the breakfast. Energy intake at lunch and during the snacking period was similar after ingesting the two sugars, while it was 15% lower after ingesting D-tagatose than with sucrose at supper (P < 0.05). Gastrointestinal factors such as the osmotic effects of unabsorbed D-tagatose causing distension of the gut might have mediated the acute appetite-suppressing effect. The present paper also refers to data from a preceding study in which we observed an increased self-reported energy intake after ingestion of D-tagatose compared with sucrose which, in fact, suggests a relative hyperphagic effect of D tagatose. However, self-reported food intake may be biased by selective under reporting and this subsequent study with a more controlled assessment of food intake was therefore conducted. This present study did not support any hyperphagic effect of D-tagatose, but rather suggests that D-tagatose may contribute to a reduced energy intake. PMID- 11029975 TI - Effects of isoenergetic overfeeding of either carbohydrate or fat in young men. AB - Ten pairs of normal men were overfed by 5 MJ/d for 21 d with either a carbohydrate-rich or a fat-rich diet (C- and F-group). The two subjects in each pair were requested to follow each other throughout the day to ensure similar physical activity and were otherwise allowed to maintain normal daily life. The increase in body weight, fat free mass and fat mass showed great variation, the mean increases being 1.5 kg, 0.6 kg and 0.9 kg respectively. No significant differences between the C- and F-group were observed. Heat production during sleep did not change during overfeeding. The RQ during sleep was 0.86 and 0.78 in the C- and F-group respectively. The accumulated faecal loss of energy, DM, carbohydrate and protein was significantly higher in the C- compared with the F group (30, 44, 69 and 51% higher respectively), whereas the fat loss was the same in the two groups. N balance was not different between the C- and F-group and was positive. Fractional contribution from hepatic de novo lipogenesis, as measured by mass isotopomer distribution analysis after administration of [1 (13)C]acetate, was 0.20 and 0.03 in the C-group and the F-group respectively. Absolute hepatic de novo lipogenesis in the C-group was on average 211 g per 21 d. Whole-body de novo lipogenesis, as obtained by the difference between fat mass increase and dietary fat available for storage, was positive in six of the ten subjects in the C-group (mean 332 (SEM 191)g per 21 d). The change in plasma leptin concentration was positively correlated with the change in fat mass. Thus, fat storage during overfeeding of isoenergetic amounts of diets rich in carbohydrate or in fat was not significantly different, and carbohydrates seemed to be converted to fat by both hepatic and extrahepatic lipogenesis. PMID- 11029976 TI - Nutritional research in World War 2: the Oxford Nutrition Survey and its research potential 50 years later. AB - To investigate the nutritional status of the population of the UK during the Second World War, nutritional surveys were commissioned in 1941. These included surveys of two groups of pregnant women: the first comprised 120 working-class women who were studied in the spring of 1942, and a second group of 253 women in 1944. Both groups were followed up until after delivery. Detailed biochemical assessments were performed on each subject. Our statistical analysis of the haematological data showed that nearly 25% of women from the 1942 group were deficient in protein, over 60% were deficient in Fe and vitamin A, and over 70% had severe vitamin C deficiency. The findings were reported to the Ministries of Health and Food who instigated a food supplementation policy at the end of 1942 that entitled pregnant women in the UK to extra rations of fruit, dairy produce and to a supply of cod-liver-oil tablets. A second group of 253 pregnant women were studied 15 months later which enabled the effects of this programme to be investigated. Supplementation reduced the proportion of women with vitamin A concentrations below the normal range from 63% to 38%, and vitamin C from 78% to 20%, but protein and Fe concentrations were not increased but actually declined. These findings continued to exert an influence over government food policy for pregnant women until the abolition of rationing in 1954. PMID- 11029977 TI - Functional foods: cholesterol-lowering benefits of plant sterols. PMID- 11029978 TI - Rheumatic diseases--surgical treatment. A systematic literature review by SBU- the Swedish Council on Technology Assessment in Health Care. PMID- 11029979 TI - Elections and public health. PMID- 11029980 TI - When is a disease eradicable? 100 years of lessons learned. AB - Since the 1915 launch of the first international eradication initiative targeting a human pathogen, much has been learned about the determinants of eradicability of an organism. The authors outline the first 4 eradication efforts, summarizing the lessons learned in terms of the 3 types of criteria for disease eradication programs: (1) biological and technical feasibility, (2) costs and benefits, and (3) societal and political considerations. PMID- 11029981 TI - Measles eradication: is it in our future? AB - Measles eradication would avert the current annual 1 million deaths and save the $1.5 billion in treatment and prevention costs due to measles in perpetuity. The authors evaluate the biological feasibility of eradicating measles according to 4 criteria: (1) the role of humans in maintaining transmission, (2) the availability of accurate diagnostic tests, (3) the existence of effective vaccines, and (4) the need to demonstrate elimination of measles from a large geographic area. Recent successes in interrupting measles transmission in the United States, most other countries in the Western Hemisphere, and selected countries in other regions provide evidence for the feasibility of global eradication. Potential impediments to eradication include (1) lack of political will in some industrialized countries, (2) transmission among adults, (3) increasing urbanization and population density, (4) the HIV epidemic, (5) waning immunity and the possibility of transmission from subclinical cases, and (6) risk of unsafe injections. Despite these challenges, a compelling case can be made in favor of measles eradication, and the authors believe that it is in our future. The question is when. PMID- 11029983 TI - Cholera in Paris. PMID- 11029982 TI - Childhood mortality impact and costs of integrating vitamin A supplementation into immunization campaigns. AB - Country-specific activity and coverage data were used to estimate the childhood mortality impact (deaths averted) and costs of integrating vitamin A supplements into immunization campaigns conducted in 1998 and 1999. More than 94 million doses of vitamin A were administered in 41 countries in 1998, helping to avert nearly 169,000 deaths. During 1999, delivery of more than 97 million doses in 50 countries helped avert an estimated 242,000 deaths. The estimated incremental cost per death averted was US$72 (range: 36-142) in 1998 and US$64 (range: 32 126) in 1999. The estimated average total cost of providing supplementation per death averted was US$310 (range: 157-609) in 1998 and US$276 (range: 139-540) in 1999. Costs per death averted varied by campaign, depending on the number and proportion of the child population reached, number of doses received per child, and child mortality rates. PMID- 11029984 TI - Globalization, states, and the health of indigenous peoples. AB - The consequences of globalization are mixed, and for the indigenous peoples of poor countries globalization has potentially important benefits. These are the result not of participation in the global economy but of participation in global networks of other indigenous peoples, environmental activists, and nongovernmental organizations. Since World War II, nonstate actors such as these have gained standing in international forums. It is indigenous peoples' growing visibility and ability to mobilize international support against the policies of their own national governments that has contributed in some important instances to their improved chances of survival. PMID- 11029985 TI - Implementing a syphilis elimination and importation control strategy in a low incidence urban area: San Diego County, California, 1997-1998. AB - OBJECTIVES: This study assessed a strategy designed to contain imported cases of syphilis and prevent reestablishment of ongoing transmission. METHODS: Reported syphilis cases during an endemic period (1990-1992) and an elimination period (1997-1998) were compared in San Diego, Calif. The elimination strategy, which focuses on rapid reporting of infectious syphilis cases by clinicians, prompt partner and sexual network management, outreach to marginalized populations, and implementation of an outbreak containment plan, was evaluated. RESULTS: Infectious syphilis incidence rates declined from 18.3 per 100,000 in 1998 to 1.0 per 100,000 in 1998. Of the 46 cases involving probable infection during 1997 1998, 19 (41%) were imported, mostly (79%) from Mexico. Outbreak containment procedures were implemented successfully for 2 small clusters. Outreach workers provided sexually transmitted disease information to a large number of individuals; however, no cases of infectious syphilis were identified, suggesting that syphilis transmission was not occurring among marginalized groups. CONCLUSIONS: This syphilis elimination and importation control strategy will require monitoring and adjustments. Controlling syphilis along the US-Mexico border is a necessary component of syphilis elimination in the United States. PMID- 11029986 TI - Strategies to eradicate rubella in the English-speaking Caribbean. AB - OBJECTIVE: This report presents the strategies used to eradicate rubella in the Caribbean region and the challenges faced by that effort. METHODS: Using the surveillance system for measles cases that was instituted in all countries in the Caribbean Community (CARICOM), 12 countries confirmed cases of rubella between 1992 and 1996. Rubella infections occurred in epidemic proportions in 6 countries during that period. RESULTS: On the basis of the rubella prevalence data, rubella congenital rubella syndrome (CRS) cost-benefit analysis, and cost-effectiveness of the mass campaign, the Council for Human and Social Development of CARICOM resolved, on April 21, 1998, that every effort would be made to eradicate rubella, as well as to prevent the occurrence of new cases of CRS by the end of 2000. Using the Pan American Health Organization's template for measles eradication, CARICOM proposed and implemented the main strategies for rubella and CRS eradication, and rubella mass campaigns were conducted in 18 countries. The target population, which included males and females (aged 20-40 years), was approximately 2.2 million. CONCLUSION: The major challenges for rubella eradication are attaining high vaccine coverage in the adult population and maintaining an effective surveillance system able to detect rubella activity. PMID- 11029987 TI - Toward elimination of Haemophilus influenzae type B carriage and disease among high-risk American Indian children. AB - OBJECTIVES: This report describes the epidemiology of Haemophilus influenzae type b (Hib) invasive disease and oropharyngeal colonization among Navajo and White Mountain Apache children younger than 7 years in an era of widespread immunization. METHODS: We conducted active surveillance for invasive H influenzae disease from 1992 to 1999 and an oropharyngeal carriage study from 1997 to 1999. The predominant vaccine used was PedvaxHib. RESULTS: The average annual incidence of invasive Hib disease among children younger than 24 months was 22 cases per 100,000. Of 381 children younger than 7 years, only 1 (0.3%; 95% confidence interval = 0.0%, 1.3%) was colonized with Hib; 370 (97%) had received 2 or more doses of Hib conjugate vaccine. CONCLUSIONS: Among Navajo and White Mountain Apache children, Hib conjugate vaccines have led to a sustained reduction in invasive Hib disease and a reduction in oropharyngeal Hib carriage. The disease incidence among children younger than 24 months remains 20 times higher than in the general US population. Hib elimination will require additional characterization of colonization and disease in these high-risk populations. PMID- 11029988 TI - Unseen blindness, unheard deafness, and unrecorded death and disability: congenital rubella in Kumasi, Ghana. AB - OBJECTIVES: Although rubella serosusceptibility among women of reproductive age in West Africa ranges from 10% to 30%, congenital rubella syndrome has not been reported. In Ghana, rubella immunization and serologic testing are unavailable. Our objectives were to identify congenital rubella syndrome cases, ascertain rubella antibody seroprevalence during pregnancy, and recommend strategies for congenital rubella syndrome surveillance. METHODS: Congenital rubella syndrome cases were identified through prospective surveillance and retrospective surveys of hospital records. A rubella serosurvey of pregnant urban and rural women was performed. RESULTS: Eighteen infants born within a 5-month period met the congenital rubella syndrome case definitions, coinciding with a 9-fold increase in presentation of infantile congenital cataract. The congenital rubella syndrome rate for this otherwise unrecorded rubella epidemic was conservatively estimated to be 0.8 per 1000 live births. A postepidemic rubella immunity rate of 92.6% was documented among 405 pregnant women; susceptibility was significantly associated with younger age (P = .000) and ethnicity (northern tribes, P = .024). CONCLUSIONS: Congenital rubella syndrome occurs in Ghana but is not reported. Information about congenital rubella syndrome and rubella in sub-Saharan Africa is needed to evaluate inclusion of rubella vaccine in proposed measles control campaigns. PMID- 11029989 TI - Estimating future hepatitis C morbidity, mortality, and costs in the United States. AB - OBJECTIVES: This study estimated future morbidity, mortality, and costs resulting from hepatitis C virus (HCV). METHODS: We used a computer cohort simulation of the natural history of HCV in the US population. RESULTS: From the year 2010 through 2019, our model projected 165,900 deaths from chronic liver disease, 27,200 deaths from hepatocellular carcinoma, and $10.7 billion in direct medical expenditures for HCV. During this period, HCV may lead to 720,700 years of decompensated cirrhosis and hepatocellular carcinoma and to the loss of 1.83 million years of life in those younger than 65 at a societal cost of $21.3 and $54.2 billion, respectively. In sensitivity analysis, these estimates depended on (1) whether patients with HCV and normal transaminase levels develop progressive liver disease, (2) the extent of alcohol ingestion, and (3) the likelihood of dying from other causes related to the route of HCV acquisition. CONCLUSIONS: Our results confirm prior Centers for Disease Control and Prevention projections and suggest that HCV may lead to a substantial health and economic burden over the next 10 to 20 years. PMID- 11029990 TI - Association of hepatitis B virus infection with other sexually transmitted infections in homosexual men. Omega Study Group. AB - OBJECTIVES: This study determined the prevalence and factors associated with hepatitis B virus (HBV) infection among men who have sex with men. METHODS: At the baseline visit of an HIV study among men who have sex with men, we asked about HBV vaccination status and tested for HBV markers. RESULTS: Of 625 subjects, 48% had received at least 1 dose of HBV vaccine. Of 328 unvaccinated men, 41% had 1 or more HBV markers. HBV prevalence increased markedly with age and was associated with many sexual and drug-related behaviors. In a multivariate model, 7 variables were independently associated with HBV infection: ulcerative sexually transmitted diseases (odds ratio [OR] = 10.1; 95% confidence interval [CI] = 2.6, 54); injection drug use (OR = 5.2; 95% CI = 1.2, 26); gonorrhea or chlamydia (OR = 4.0; 95% CI = 1.9, 8.9); sexual partner with HIV/AIDS (OR = 3.6; 95% CI = 1.8, 7.1); 50 or more casual partners (OR = 3.4; 95% CI = 1.6, 7.1); received money for sex (OR = 3.0; 95% CI = 1.2, 7.8); and 20 or more regular partners (OR = 2.5; 95% CI = 1.1, 6.1). CONCLUSIONS: In Montreal, men who have sex with men are at risk for HBV infection, but a substantial proportion remain unvaccinated; new strategies are required to improve coverage. Men who have sex with men and who have a sexually transmitted infection, especially a genito ulcerative infection, appear to be at particularly high risk for HBV infection. PMID- 11029991 TI - Driver air bag effectiveness by severity of the crash. AB - OBJECTIVES: This analysis provided effectiveness estimates of the driver-side air bag while controlling for severity of the crash and other potential confounders. METHODS: Data were from the National Automotive Sampling System (1993-1996). Injury severity was described on the basis of the Abbreviated Injury Scale, Injury Severity Score, Functional Capacity Index, and survival. Ordinal, linear, and logistic multivariate regression methods were used. RESULTS: Air bag deployment in frontal or near-frontal crashes decreases the probability of having severe and fatal injuries (e.g., Abbreviated Injury Scale score of 4-6), including those causing a long-lasting high degree of functional limitation. However, air bag deployment in low-severity crashes increases the probability that a driver (particularly a woman) will sustain injuries of Abbreviated Injury Scale level 1 to 3. Air bag deployment exerts a net injurious effect in low severity crashes and a net protective effect in high-severity crashes. The level of crash severity at which air bags are protective is higher for female than for male drivers. CONCLUSIONS: Air bag improvement should minimize the injuries induced by their deployment. One possibility is to raise their deployment level so that they deploy only in more severe crashes. PMID- 11029992 TI - The association of sexual behaviors with socioeconomic status, family structure, and race/ethnicity among US adolescents. AB - OBJECTIVES: This study assessed the relation of socioeconomic status (SES), family structure, and race/ethnicity to adolescent sexual behaviors that are key determinants of pregnancy and sexually transmitted diseases (STDs). METHODS: The 1992 Youth Risk Behavior Survey/Supplement to the National Health Interview Survey provided family data from household adults and behavioral data from adolescents. RESULTS: Among male and female adolescents, greater parental education, living in a 2-parent family, and White race were independently associated with never having had sexual intercourse. Parental education did not show a linear association with other behaviors. Household income was not linearly related to any sexual behavior. Adjustment for SES and family structure had a limited effect on the association between race/ethnicity and sexual behaviors. CONCLUSIONS: Differences in adolescent sexual behavior by race and SES were not large enough to fully explain differences in rates of pregnancy and STD infection. This suggests that other factors, including access to health services and community prevalence of STDs, may be important mediating variables between SES and STD transmission and pregnancy among adolescents. PMID- 11029993 TI - The alcohol warning and adolescents: 5-year effects. AB - OBJECTIVES: This study, a follow-up to the authors earlier report, examined the effects of the alcohol warning label on adolescents during the first 5 years that the warning was required. METHODS: Surveys were administered to 10th-grade (n = 16,661) and 12th-grade (n = 15,856) students from the 1989-1990 school year through the 1994-1995 school year. The measures were awareness of, exposure to, and recognition memory of the alcohol warning label; beliefs about the risks listed on the warning; and open-ended statements about consequences of alcohol use, alcohol consumption, and self-reported driving after drinking. RESULTS: There were increases in warning awareness, exposure, and recognition memory. These effects leveled off approximately 3.5 years after the inclusion of the warning on alcohol beverage containers. There was no beneficial change attributable to the warning in beliefs, alcohol consumption, or driving after drinking. CONCLUSIONS: The initial positive effects of the alcohol warning label on adolescents have leveled off, consistent with theories of repeated exposure to persuasive information. The alcohol warning has not affected adolescents' beliefs about alcohol or alcohol-related behaviors. PMID- 11029994 TI - Knowledge, attitudes, and screening practices among older men regarding prostate cancer. AB - OBJECTIVES: This study determined population-based rates of reported prostate cancer screening and assessed prostate cancer-related knowledge, attitudes, and screening practices among men in New York aged 50 years and older. METHODS: Two telephone surveys were conducted. One was included in the 1994 and 1995 statewide Behavioral Risk Factor Surveillance System interviews, and the other was a community-level survey that targeted Black men (African-American Men Survey). Prevalence estimates were computed for each survey, and prostate cancer screening practices were assessed with logistic regression models. RESULTS: Overall, fewer than 10% of the men in each survey perceived their prostate cancer risk to be high; almost 20% perceived no risk of developing the disease. Approximately 60% of the men in each survey reported ever having had a prostate-specific antigen (PSA) test. In both surveys, physician advice was significantly associated with screening with a PSA test or a digital rectal examination. Also, race was significantly associated with screening in the statewide survey. CONCLUSIONS: Many New York men appear to be unaware of risk factors for prostate cancer. However, a substantial percentage reported having been screened for the disease; physician advice may have been a major determining factor in their decision to be tested. PMID- 11029995 TI - Cancer incidence and survival following bereavement. AB - OBJECTIVES: This study investigated the effect of parental bereavement on cancer incidence and survival. METHODS: A cohort of 6284 Jewish Israelis who lost an adult son in the Yom Kippur War or in an accident between 1970 and 1977 was followed for 20 years. We compared the incidence of cancer in this cohort with that among nonbereaved members of the population by logistic regression analysis. The survival of bereaved parents with cancer was compared with that of matched controls with cancer. RESULTS: Increased incidence was found for lymphatic and hematopoietic malignancies among the parents of accident victims (odds ratio [OR] = 2.01; 95% confidence interval [CI] = 1.30, 3.11) and among war-bereaved parents (OR = 1.47; 95% CI = 1.13, 1.92), as well as for melanomas (OR = 4.62 [95% CI = 1.93, 11.06] and 1.71 [95% CI = 1.06, 2.76], respectively). Accident-bereaved parents also had an increased risk of respiratory cancer (OR = 1.50; 95% CI = 1.07, 2.11). The survival study showed that the risk of death was increased by bereavement if the cancer had been diagnosed before the loss, but not after. CONCLUSIONS: This study showed an effect of stress on the incidence of malignancies for selected sites and accelerated demise among parents bereaved following a diagnosis of cancer, but not among those bereaved before such a diagnosis. PMID- 11029996 TI - The economic implications of self-care: the effect of lifestyle, functional adaptations, and medical self-care among a national sample of Medicare beneficiaries. AB - OBJECTIVES: Self-care includes actions taken by individuals to promote or ensure their health, to recover from diseases or injuries, or to manage their effects. This study measured associations between self-care practices (lifestyle practices, adaptations to functional limitations, and medical self-care) and Medicare expenditures among a national sample of adults 65 years and older. METHODS: Regression models of Medicare use and expenditures were estimated by using the National Survey of Self-Care and Aging and Medicare claims for 4 years following a baseline interview. RESULTS: Lifestyle factors (swimming and walking) and functional adaptations (general home modifications) were associated with reductions in monthly Medicare expenditures over a 12-month follow-up period. Expenditure reductions were found over the 48-month follow-up period for participation in active sports, gardening, and medical self-care. Practices associated with increases in expenditures included smoking, physical exercise (possibly of a more strenuous nature), and specific home modifications. CONCLUSIONS: Certain self-care practices appear to have significant implications for Medicare expenditures and presumptively for the health status of older adults. Such practices should be encouraged among older adults as a matter of national health policy. PMID- 11029997 TI - Determining immunization rates for inner-city infants: statewide registry data vs medical record review. AB - OBJECTIVE: This study evaluated the benefit of consulting a statewide immunization registry for inner-city infants whose immunizations appeared, after single-site chart review, to have been delayed. METHODS: We prospectively enrolled 315 newborns in 3 inner-city pediatric clinics. When the infants turned 7 months old, we obtained immunization data from clinic charts and the state registry. RESULTS: On the basis of chart review, 147 infants (47%) were assessed to be delayed in their immunizations; of these, registry data revealed that 28 (19%) had received additional immunizations and 15 (10%) were actually up to date. CONCLUSIONS: A statewide registry can capture immunizations from multiple sources, improving accurate determination of immunization rates in a mobile, inner-city population. PMID- 11029998 TI - Risk and prevalence of treatable sexually transmitted diseases at a Birmingham substance abuse treatment facility. AB - OBJECTIVES: We evaluated the prevalence of gonorrhea, chlamydia, trichomoniasis, and syphilis in patients entering residential drug treatment. METHODS: Data on sexual and substance abuse histories were collected. Participants provided specimens for chlamydia and gonorrhea ligase chain reaction testing. Trichomonas vaginalis culture, and syphilis serologic testing. RESULTS: Of 311 patients, crack cocaine use was reported by 67% and multisubstance use was reported by 71%. Sexually transmitted disease (STD) risk behaviors were common. The prevalence of infection was as follows: Chlamydia trachomatis, 2.3%; Neisseria gonorrhoeae, 1.6%; trichomoniasis, 43%; and syphilis, 6%. CONCLUSIONS: STD counseling and screening may be a useful adjunct to inpatient drug treatment. PMID- 11029999 TI - The Safe Dates program: 1-year follow-up results. AB - OBJECTIVES: An earlier report described desirable 1-month follow-up effects of the Safe Dates program on psychological, physical, and sexual dating violence. Mediators of the program-behavior relationship also were identified. The present report describes the 1-year follow-up effects of the Safe Dates program. METHODS: Fourteen schools were in the randomized experiment. Data were gathered by questionnaires in schools before program activities and 1 year after the program ended. RESULTS: The short-term behavioral effects had disappeared at 1 year, but effects on mediating variables such as dating violence norms, conflict management skills, and awareness of community services for dating violence were maintained. CONCLUSIONS: The findings are considered in the context of why program effects might have decayed and the possible role of boosters for effect maintenance. PMID- 11030000 TI - Put prevention into practice: a controlled evaluation. AB - OBJECTIVES: The purpose of this study was to evaluate whether Put Prevention Into Practice (PPIP) materials affected the delivery of 8 clinical preventive services. METHODS: Program materials were provided to a family medicine practice serving a diverse, low-income population. Appropriate use of clinical preventive services was assessed via medical record reviews at baseline, 6 months, 18 months, and 30 months at both intervention and control sites. RESULTS: The delivery rates of 7 clinical preventive services were higher in the intervention site at 6 months. These rates had flattened or decreased by 30 months. CONCLUSIONS: Use of PPIP materials modestly improved delivery of certain clinical preventive services. Sustained improvement will require substantial system changes and ongoing support. PMID- 11030001 TI - Physician firearm ownership as a predictor of firearm injury prevention practice. AB - OBJECTIVES: This study explored the relation between physicians' gun ownership and their attitudes and practices regarding firearm injury prevention. METHODS: Internists and surgeons were surveyed, and logistic regression models were developed with physicians' personal involvement with firearms (in the form of a gun score) as the primary independent variable. RESULTS: Higher gun scores were associated with less agreement that firearm injury is a public health issue and that physicians should be involved in firearm injury prevention but with a greater likelihood of reporting the inclusion of firearm ownership and storage as part of patient safety counseling. CONCLUSIONS: Despite being less likely to say that doctors should participate in firearm injury prevention, physician gun owners are more likely than nonowners to report counseling patients about firearm safety. PMID- 11030002 TI - Hunger in legal immigrants in California, Texas, and Illinois. AB - OBJECTIVES: This study determined the prevalence of food insecurity and hunger in low-income legal immigrants. METHODS: We conducted a cross-sectional survey of Latino and Asian legal immigrants attending urban clinics and community centers in California, Texas, and Illinois with a food security questionnaire. RESULTS: Among 630 respondents, 40% of the households were food insecure without hunger and 41% were food insecure with hunger. Independent predictors of hunger were income below federal poverty level (odds ratio [OR] = 2.72, 95% confidence interval [CI] = 1.72, 4.30), receipt of food stamps (OR = 2.53, 95% CI = 1.57, 4.09), Latino ethnicity (OR = 2.39, 95% CI = 1.49, 3.82), and poor English (OR = 1.76, 95% CI = 1.10, 2.82). CONCLUSIONS: The prevalence of hunger among low income legal immigrants is unacceptably high. Access to food assistance programs is important for the health and well-being of this population. PMID- 11030003 TI - Endemic goiter associated with high iodine intake. AB - OBJECTIVES: This study assessed the relation of iodine content of household water to thyroid size and urinary iodine excretion in an area with high iodine concentration in the water. METHODS: The iodine content of household water and indicators of iodine status (thyroid size and urinary iodine level) were assessed in selected villages in Jiangsu Province, China. RESULTS: Water iodine levels were positively correlated with urinary iodine levels and indicators of thyroid size at the township level. CONCLUSIONS: Excess iodine in household water was the likely cause of endemic goiter and elevated urinary iodine levels in the study area. This finding affects public health policy on the institution of universal salt iodization for the elimination of iodine deficiency disorders. PMID- 11030005 TI - Nonprescription pharmacy sales of needles and syringes. PMID- 11030004 TI - The association of dietary folate, B6, and B12 with cardiovascular mortality in Spain: an ecological analysis. AB - OBJECTIVES: This study assessed the association of dietary folate, vitamin B6, and vitamin B12 with cardiovascular mortality. METHODS: Poisson regression analyses assessed coronary/cerebrovascular mortality rates via nutrient data obtained from the National Nutrition Survey, which recorded 7-day food intakes from a national sample of 21,155 households. RESULTS: In regard to coronary mortality, male and female rate ratios (highest vs lowest quintile) were 0.83 (95% confidence interval [CI] = 0.77, 0.91) and 0.95 (95% CI = 0.86, 1.05), respectively, for folate and 0.74 (95% CI = 0.65, 0.84) and 0.86 (95% CI = 0.73, 0.99), respectively, for B12. Intake of folate and B6 (but not B12) was significantly associated with cerebrovascular mortality. CONCLUSIONS: B vitamins are associated with cardiovascular mortality in the general population. PMID- 11030006 TI - Optimal immunization practices for the special supplemental nutrition program for women, infants, and children. PMID- 11030007 TI - An outbreak of Shigella sonnei associated with a recreational spray fountain. PMID- 11030008 TI - Domestic violence and HIV/AIDS. PMID- 11030010 TI - Index volumes 104-113. PMID- 11030009 TI - VIII International Congress of the Metastasis Research Society. London, United Kingdom, 24-27 September 2000. Abstracts. PMID- 11030011 TI - New York Academy of Dentistry member directory. PMID- 11030012 TI - 3rd International Workshop on Salvage Therapy for HIV Infection. 12-14 April 2000, Chicago, Illinois, USA. Report and Abstracts. PMID- 11030013 TI - The 4th annual scientific meeting of the Heart Failure Society of America. September 10-13, 2000. Abstracts. PMID- 11030015 TI - 18th World Congress on Endourology and SWL 16th Basic Research Symposium. September 14-17, 2000. Sao Paulo, Brazil. Abstracts. PMID- 11030014 TI - American Association of Cardiovascular and Pulmonary Rehabilitation annual meeting, Tampa, Florida, USA, September 21-24, 2000. Canadian Association of Cardiac Rehabilitation, 10th annual meeting, Burnaby, British Columbia, Canada, October 27-29, 2000. Abstracts. PMID- 11030016 TI - Overview of the angiotensin-converting-enzyme inhibitors. AB - The pharmacology of angiotensin-converting-enzyme (ACE) inhibitors and their role in the renin-angiotensin system (RAS) are described, and pharmacokinetic properties and common adverse events are presented. ACE inhibitors play a vital role in the RAS by regulating the potent vasoconstrictor angiotensin II. All ACE inhibitors share the same basic structure; however, they can be separated on the basis of their functional (binding) group: carboxyl, sulfhydryl, or phosphinyl. These functional groups are, in part, responsible for differences in the pharmacokinetic and safety profiles of these agents. Captopril and lisinopril are the only ACE inhibitors that are not prodrugs requiring activation through hepatic biotransformation. Differences among the ACE inhibitors in lipophilicity are described; fosinopril has the greatest lipophilicity and lisinopril the least. ACE is found in numerous tissues, and there is increasing evidence of differences among ACE inhibitors in their ability to inhibit tissue ACE. Most ACE inhibitors are eliminated mainly by the kidneys and to a lesser extent through the liver. Lisinopril is the only ACE inhibitor that does not require hepatic metabolism. In the selection of an ACE inhibitor for once-daily use to treat hypertension, differences in trough-peak ratios are clinically relevant. Fosinopril, ramipril, and trandolapril have minimum trough-peak ratios of 50% or greater. ACE inhibitors are generally well tolerated, with hypotension, cough, and hyperkalemia being the most frequently reported adverse effects for the entire class. Drug interactions across the ACE inhibitor class as well as agent specific interactions are described. Factors to be considered in the selection of an ACE inhibitor include differences in potency, affinity for ACE, pharmacokinetics, and toxicity that are related to structural properties of the drug; whether the trough-peak ratio enables use of a once-daily dose; and potential adverse effects related to a drug's functional (binding) group. PMID- 11030017 TI - Role of angiotensin-converting-enzyme inhibitors in the treatment of hypertension. AB - Therapeutic goals for the treatment of hypertension and the ability of various angiotensin-converting-enzyme (ACE) inhibitors to meet these goals are presented. The 1997 Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC-VI) does not recommend ACE inhibitors for first-line therapy in the treatment of hypertension; however, these guidelines do identify compelling indications for ACE inhibitor therapy, including diabetes mellitus (type 1) with proteinuria, heart failure, or previous myocardial infarction with systolic dysfunction. Since the JNC-VI guidelines were developed, the results of a prospective randomized clinical trial in patients with uncomplicated hypertension have demonstrated that ACE inhibitor therapy is as effective as conventional treatment in the prevention of cardiovascular morbidity and mortality. In hypertensive patients with diabetes, therapy with captopril, enalapril, fosinopril, or ramipril has resulted in significant reductions in cardiovascular events. In addition, tight blood pressure control with an ACE inhibitor has resulted in a greater reduction in the risk of macrovascular and microvascular complications of diabetes than was seen with less tight control. Recent study results support broader use of ACE inhibitors for hypertension than was recommended in the JNC-VI guidelines. PMID- 11030018 TI - Role of angiotensin-converting-enzyme inhibition in patients with renal disease. AB - Clinical studies evaluating the benefits of angiotensin-converting-enzyme (ACE) inhibitor therapy in patients likely to develop renal disease are reviewed. Patients with diabetes or hypertension are at increased risk for development of renal disease. In patients with diabetic nephropathy, captopril therapy was associated with a 50% reduction in the risk of death, dialysis, and transplantation and a significantly smaller increase in serum creatinine compared with placebo. Therapy with enalapril or lisinopril has been shown to limit the progression of renal disease in normoalbuminuric patients with diabetes. Long term therapy with enalapril (up to seven years) has demonstrated the ability to preserve renal function in patients with diabetes and microalbuminuria. Over 4.5 years, patients with diabetes and at least one other cardiovascular risk factor had significant reductions in the risk of overt nephropathy with ramipril therapy compared with placebo. In addition, ramipril is associated with preservation of renal function in patients with nondiabetic nephropathy. Evidence suggesting a dissociation of the renal hemodynamic and antiproteinuric effects of ACE inhibition is presented. These positive effects of ACE inhibition cannot be explained by reductions in blood pressure alone. ACE inhibitors have renoprotective properties beyond systemic blood pressure reduction. PMID- 11030019 TI - Angiotensin-converting-enzyme inhibition in heart failure or after myocardial infarction. AB - The use of angiotensin-converting-enzyme (ACE) inhibitors in patients with heart failure and after myocardial infarction (MI) is discussed, and results of relevant studies are reviewed. In several large trials, the administration of captopril or lisinopril within the first 36 hours after the onset of chest pain due to MI was associated with significant reductions in mortality risk, compared with placebo. Trials evaluating the use of captopril, ramipril, or trandolapril at least three days after the onset of chest pain due to MI also demonstrated significant reductions in mortality risk. Pivotal clinical trials of captopril, enalapril, lisinopril, and ramipril in the treatment of heart failure are presented. Overall, ACE inhibitor therapy was shown to reduce mortality by decreasing the progression of heart failure. Possible benefits of ACE inhibition in addition to reductions in afterload and preload and preservation of serum potassium are discussed. Certain ACE inhibitors may exert positive effects by modulating plasminogen activator inhibitor-1, endothelial function, and left ventricular remodeling. If not contraindicated, long-term therapy with captopril, lisinopril, or ramipril should be used in post-MI patients. Patients with heart failure should be treated with one of the ACE inhibitors that have been shown beneficial for this indication. PMID- 11030020 TI - Factors to consider in selecting an angiotensin-converting-enzyme inhibitor. AB - The results of clinical studies of the use of angiotensin-converting-enzyme (ACE) inhibitors in a variety of cardiovascular and renal disorders are reviewed, with emphasis on evidence-based formulary decision-making. In evaluating agents for formulary inclusion, efficacy, safety, dosing, and cost are considered. ACE inhibitors have been investigated in thousands of patients in clinical trials, so an evidence-based approach to their use is appropriate. All ACE inhibitors are effective antihypertensive agents and are generally well tolerated. Thus, in formulary decisions, once-daily dosing and good 24-hour blood pressure control are important considerations to ensure efficacy and patient compliance. ACE inhibitors are key in the prevention and management of heart failure but are underused for these indications. The rationale for use of ACE inhibitors after myocardial infarction is discussed, as is evidence of separate antihypertensive and renoprotective effects of ACE inhibitors. Evidence for the use of ramipril in patients at high risk for cardiovascular events, from the Heart Outcomes Prevention Evaluation Study (HOPE), is reviewed. Clinical trial results provide evidence that some ACE inhibitors have renoprotective and vasculoprotective effects and improve outcomes of patients with a variety of cardiovascular disorders; differences among the ACE inhibitors should be considered in formulary decision-making. PMID- 11030021 TI - FDA draws patients into alosetron risk management. PMID- 11030023 TI - Formulary system principles distributed to policymakers. PMID- 11030022 TI - Unithroid becomes first levothyroxine tablet to gain FDA approval. PMID- 11030024 TI - Nursing homes may have 350,000 adverse drug events annually, investigators suggest. PMID- 11030025 TI - New resuscitation guidelines released. PMID- 11030026 TI - Risks of using technicians and not pharmacists to handle antineoplastic drugs. PMID- 11030027 TI - Pharmacy's changing role in dispensing and administration. PMID- 11030028 TI - ASHP national survey of pharmacy practice in acute care settings: dispensing and administration--1999. AB - Results of the 1999 ASHP national survey of pharmacy practice in acute care settings that pertain to drug dispensing and administration practices are presented. Pharmacy directors at 1050 general and children's medical-surgical hospitals in the United States were surveyed by mail. The response rate was 51%. About three-fourths of respondents described their inpatient pharmacy's distribution system as centralized. Of those with centralized distribution, 77.4% indicated that their system was not automated. Decentralized pharmacists were used in 29.4% of the hospitals surveyed; an average of 58.9% of their time was spent on clinical, as opposed to distributive, activities. About 67% of directors reported pharmacy computer access to hospital laboratory data, 38% reported access to automated medication-dispensing-unit data, and 19% reported computer access to hospital outpatient affiliates. Only 13% of hospitals had an electronic medication order-entry system; another 27% reported they were in the process of developing such a system. Decentralized medication storage and distribution devices were used in 49.2% of hospitals, while 7.3% used bedside information systems for medication management. Machine-readable coding was used for inpatient pharmacy dispensing by 8.2% of hospitals. Ninety percent reported a formal, systemwide committee responsible for data collection, review, and evaluation of medication errors. Virtually all respondents (98.7%) reported that their staff initiated manual reports. Only two thirds tracked these reports and reported trends to the staff. Fewer than 15% reported that staff were penalized for making or contributing to an error. Pharmacists are making a significant contribution to the safety of medication distribution and administration. The increased use of technology to improve efficiency and reduce costs will require that pharmacists continue to focus on the impact of changes on the safety of the medication-use system. PMID- 11030029 TI - Bioequivalence of a propylene glycol-based liquid dapsone preparation and dapsone tablets. AB - The bioequivalence of a proprietary liquid dapsone preparation and commercially available dapsone tablets was studied. Twelve adult volunteers received dapsone doses with 8 oz of water one to two hours after their usual breakfast. Each subject received an initial 100-mg dose of a propylene glycol-based liquid preparation of dapsone and, two weeks later, a 100-mg dapsone tablet (both from Jacobus Pharmaceutical Company, Princeton, NJ). Blood samples were collected before and at intervals up to 96 hours after the administration of each dose. Serum dapsone concentrations were determined by high-performance liquid chromatography, and pharmacokinetic values were calculated by model-independent analysis. The area under the concentration-versus-time curve and the maximum serum concentration for the two formulations met the criteria for bioequivalence. Time to maximum serum concentration tended to be lower for the liquid, but not significantly. The liquid and tablet formulations of dapsone studied were found to be bioequivalent and may be used interchangeably. PMID- 11030030 TI - Impact of interventions designed to increase market share and prescribing of fexofenadine at HMOs. AB - The impact of interventions designed to shift prescribing from loratadine to fexofenadine at HMOs was studied. Pharmacy claims data for a six-month preintervention period at four HMOs were analyzed to identify all new and refill prescriptions for loratadine, fexofenadine, astemizole, and cetirizine. The interventions consisted of a mandatory lockout of loratadine in favor of fexofenadine (at HMO A), a voluntary switch to fexofenadine promoted through letters to both physicians and members (HMO B), and a voluntary switch promoted through letters to physicians only (HMO C). There was no intervention at HMO D. Pharmacy claims data for the six months after each intervention program was implemented were analyzed to determine changes in the market share and prescribing of the study drugs. After the intervention programs were implemented, the market share of fexofenadine increased from 18.9% to 65.2% at HMO A, from 14.8% to 21.0% at HMO B, and from 20.7% to 23.8% at HMO C. Loratadine's market share decreased from 62.3% to 8.7% at HMO A, from 67.5% to 58.6% at HMO B, and from 70.5% to 65.3% at HMO C. HMOs A, B, and C each had greater shifts in market share for fexofenadine and loratadine than the control HMO. Changes in prescribing followed a similar pattern for the 25 physicians at each HMO who had most frequently prescribed loratadine during the preintervention period. The average cost per antihistamine prescription decreased 22.3% at HMO A. Prescription costs continued to rise at HMOs B, C, and D. Mandating the use of fexofenadine produced a significant increase in its market share, reduced the cost of nonsedating antihistamines, and successfully influenced prescribing behavior. Voluntary programs had a more modest impact on market share and did not stop increases in prescription costs. PMID- 11030031 TI - Development of a pharmacy residency program in home care. AB - A residency training program developed by a college of pharmacy in conjunction with a home care company is described. The 12-month program is based on the ASHP Residency Learning System and the goal statements and educational objectives of ASHP's accreditation standard for pharmacy practice residency training with emphasis in home care. Establishing the program involved identifying goals, objectives, and learning experiences consistent with the expected outcomes. Specific objectives for meeting goals in four categories--practice foundation skills, direct patient care, drug information and drug policy development, and practice management--were linked to expected program outcomes. Learning experiences that would lead to achievement of the program objectives and outcomes were selected and organized into one- to eight-week rotations (e.g., in acute care, care of pediatric patients, pain management, nutrition, patient education and counseling, and administration and practice management). Throughout the program, residents gain experience in pharmaceutical services and research. Skills in care planning and monitoring are emphasized, as is practicing pharmaceutical care in an interdisciplinary environment. Residents who have completed the program have found employment immediately as pharmacy managers of home infusion centers. Pharmacy residency training in home care provides the experience needed to function as a competent clinician and manager who can identify and solve problems to improve patient care. PMID- 11030032 TI - Levofloxacin use at an academic teaching institution. PMID- 11030033 TI - The habits of successful pharmacists. PMID- 11030034 TI - Therapeutic interchange of low-molecular-weight heparins. PMID- 11030035 TI - Therapeutic interchange of low-molecular-weight heparins. PMID- 11030036 TI - Therapeutic interchange of low-molecular-weight heparins. PMID- 11030037 TI - Interaction potential of selective serotonin-reuptake inhibitors. PMID- 11030039 TI - Stem cell transplantation for non-malignant disorders. AB - Stem cell transplantation (SCT) can be used to cure or ameliorate a wide variety of non-malignant diseases. These range from inherent defects of haemopoietic cell production or function, through metabolic diseases (where blood cells are providing in vivo enzyme therapy to solid organs), to severe autoimmune diseases. However, although transplantation has revolutionized the treatment of many of the diseases discussed, severe toxicities remain. In some cases these are inherent to the disease concerned but frequently they relate to the conditioning regime or post-transplant complications such as graft-versus-host disease (GvHD). This chapter concentrates on the indications for transplant, outcome statistics and problems inherent in particular conditions, seen in the light of technological improvements during the 1990s and the potential impact of enzyme and gene therapies. PMID- 11030038 TI - The diagnosis and management of hereditary spherocytosis. AB - Hereditary spherocytosis (HS) is relatively common in Caucasian populations; most individuals have mild or only moderate disease. There is commonly a family history and a typical clinical and laboratory picture so that the diagnosis is usually easily made without additional laboratory tests. Atypical cases may require measurement of membrane proteins and molecular genetics to clarify the nature of the membrane disorder. It is particularly important to rule out stomatocytosis because splenectomy is contraindicated because of the thrombotic risk. Mild HS can be managed without folate supplements and does not require splenectomy. Moderately and severely affected individuals are likely to benefit from splenectomy, which should be performed after the age of 6 and with appropriate counselling about the risk of infection. In all cases careful dialogue between physician, child and the family is essential. Laparoscopic surgery can result in shorter hospital stay and less pain. PMID- 11030040 TI - The management of alloimmune neonatal thrombocytopenia. AB - Neonatal alloimmune thrombocytopenia (NAITP), defined as thrombocytopenia (platelet count < 150 x 10(9)/l) due to transplacentally acquired maternal platelet alloantibodies, occurs in approximately 1 per 1200 live births in a Caucasian population. In such a population, the majority (> 75 percent) of cases are due to fetomaternal incompatibility for the platelet specific alloantigen, HPA-1a (P1A1, Zwa). Incompatibility for the HPA-5b (Bra) alloantigen is the next most frequent cause of NAITP in Caucasians; much less common is NAITP due to incompatibility for HLA, blood group ABO or other platelet-specific antigens. In non-Caucasian populations (e.g. Orientals) HPA-1a incompatibility is a rare cause of NAITP and other alloantigens e.g. HPA-4b (Penb, Yuka) are implicated. The greatest clinical challenge relates to the antenatal management of pregnant women alloimmunized to the HPA-1a (P1A1, Zwa) antigen, and particularly the subset of such women who have a history of a previously affected infant with severe thrombocytopenia and/or intracranial hemorrhage (ICH). The risk of antenatal ICH in the fetus of such women is high enough to merit intervention, either weekly infusion of high-dose intravenous immunoglobulin G (IVIG) with or without corticosteroids given to the mother (the preferred approach in North American centres), or repeated in-utero fetal platelet transfusions (the preferred treatment approach in some European centres). Post-natal management of severely affected infants centres on the rapid provision of compatible antigen-negative platelets harvested from the mother or a phenotyped donor. The value of antenatal screening programs to detect 'at risk' alloimmunized women during pregnancy continues to be debated. PMID- 11030041 TI - Diamond-Blackfan anaemia. AB - Diamond-Blackfan anaemia (DBA) has had an intellectual allure for decades for clinical and experimental haematologists. The syndrome has a haematological phenotype of early-onset red-cell aplasia but is coupled with a baffling array of pleiotropy. There is discordance with modes of inheritance, physical anomalies, erythropoietic response to corticosteroid therapy, spontaneous 'remissions', and evolution to malignant myeloid transformation and to cancer. The recent discovery of two genes associated with DBA is the entry point for explaining the diversity of the phenotype and for understanding the molecular basis of the syndrome. PMID- 11030042 TI - The genetics of Fanconi's anaemia. AB - Fanconi's anaemia (FA) is an inherited bone marrow failure syndrome characterized by considerable clinical and cellular heterogeneity. This has also been recently demonstrated at the genetic and molecular levels following cloning of four out of the seven FA genes. Although this now enables molecular diagnosis in the majority of patients, because of the considerable molecular heterogeneity, the diepoxybutane/mitomycin-C stress test based on the increased chromosomal instability seen in FA cells, compared to normal controls, remains the front-line diagnostic test. This FA cell hallmark has led to the suggestion that FA may represent a defect in DNA repair although the precise function of the cloned FA genes remains unknown. Recent data suggest that they function in a novel cell pathway which has an important role in maintaining chromosome stability. The advances in the genetics of FA have already had some impact on diagnosis--for example, identification of patients with somatic mosaicism who have atypical clinical presentations--but to date they have had little impact on treatment. However, new treatments may now follow; indeed, for a number of reasons, FA may be a good candidate for haemopoietic gene therapy. PMID- 11030043 TI - The genetics of childhood acute lymphoblastic leukaemia. AB - In childhood acute lymphoblastic leukaemia (ALL) a number of genetic changes have been identified which provide diagnostic and prognostic information with a direct impact on patient management. The most significant abnormalities include the translocation, t(12;21)(p13;q22), giving rise to the ETV6/AML1 gene fusion; BCR/ABL arising from t(9;22)(q34;q11); re-arrangements of the MLL gene; the E2A/PBX1 from the t(1;19)(q23;p13); re-arrangements of MYC with the immunoglobulin genes and re-arrangements of the T cell receptor genes. Chromosomal deletions, particularly those of the short arms of chromosomes 9 and 12 and the long arm of chromosome 6, have been postulated to be the sites of tumour suppressor genes (TSG). Numerical chromosomal abnormalities are of particular importance in relation to prognosis. High hyperdiploidy (50-65 chromosomes) is associated with a good risk, whereas the outlook for patients with near haploidy (23-29 chromosomes) is extremely poor. In view of the introduction of risk-adjusted therapy into the UK childhood ALL treatment trials, an interphase FISH screening programme has been developed to reveal chromosomal abnormalities with prognostic significance in childhood ALL. PMID- 11030044 TI - Management of infection in children with bone marrow failure. AB - The development of new and often more successful regimens of treatment for childhood blood and malignant diseases has usually been associated with a parallel increase in infectious problems. This is because these successes have often, in the absence of new effective drugs, been achieved by increasing drug dose intensity to new limits. This chapter is not intended to deal with every possible infection, but rather to be a practical guide to the current management of infection. The last few years have seen major improvements in the development of haematopoietic growth factors, new antifungal agents, new antibiotics and new ways to use aminoglycosides. Attempts are being made to identify good and poor risk factors for the outcome of infection in order to facilitate shorter courses and possibly home or day care use of antimicrobial agents. In addition the different needs of children are being recognized in view of the restricted use of quinolones in this group and the different organisms and types of infection that they experience. PMID- 11030045 TI - Inhibitors in young boys with haemophilia. AB - The development of an inhibitor antibody to factor VIII (or factor IX) in a child with haemophilia presents a major challenge to the paediatric haematologist. This article provides an overview of the incidence of inhibitor development in early childhood (30-52% in boys with severe haemophilia A), genetic risk factors, detection, high titre, low titre and transient inhibitors, and management. Treatment of patients with inhibitors is time-consuming and expensive. One should make every attempt to ensure that the boy's family has an understanding of inhibitors, treatment options, and just what is being recommended for their child and what this involves. Immune tolerance induction is successful in approximately 85% of boys with factor VIII inhibitors, but in only 40-50% of those with factor IX inhibitors. For treatment of bleeding episodes in children with high-titre (> or = 5 Bethesda Units) inhibitors, therapeutic options include activated prothrombin complex concentrates (APCC), rF VIIa, and (for factor VIII inhibitors) porcine factor VIII. The advantages and disadvantages of each are discussed. Although factor IX inhibitors are far less common (occurring in 2-3% of boys with haemophilia B), approximately 50% are accompanied by the occurrence of anaphylaxis or severe allergic reactions to any factor IX-containing product. PMID- 11030046 TI - Chronic childhood idiopathic thrombocytopenic purpura. AB - Childhood idiopathic thrombocytopenic purpura (ITP) is a largely trivial disorder from which over 95% of children sooner or later recover spontaneously, and for most of whom the risks of unnecessary or ineffective therapy are arguably greater than those of the untreated disease. There are, however, a few patients who continue to have very low platelet counts and remain symptomatic for many months or years. They are rare, and they present difficult management problems. Splenectomy is probably the most effective treatment but is also the most dangerous and is not always successful. It is also irreversible. Most other regimens are either ineffective, unacceptably toxic, or both. Planning management for an individual patient requires a realistic risk:benefit appraisal, a process that is impeded by inadequate epidemiological data and a scarcity of large-scale randomized clinical trials. International collaborative studies may help in the future. PMID- 11030047 TI - [Circulation of virus and interspecies contamination in wild animals]. AB - Paradoxically, just when we have succeeded in eradicating and/or bringing under control the major viral infections (smallpox, poliomyelitis, measles) numerous viral infections are emerging in man and in animals. Changes in our social environment, technological and ecological equilibrium have facilitated this phenomenon. Furthermore, certain of these viruses have demonstrated an almost unlimited capacity to adapt genetically to environmental change. HIV has already infected 40 million individuals, but monkeypox, Ebola, simian herpes can cause epidemics with serious if not fatal outcomes. Haemorrhagic fever epidemics have resulted from human contact with Flavivirus infected rodents and insects. Paramyxoviruses and morbiliviruses can cause fatal outcomes in man and animals. And the three influenza epidemics having occurred in the 20th century all came from the type A avian reservoir. The often complex combinations of predisposing factors having facilitated the emergence of several epidemics merit further consideration. PMID- 11030048 TI - [Ecology and social organization of African tropical forest primates: aid in understanding retrovirus transmission]. AB - The risk of transmission of primate viruses to humans is great because of their genetic proximity. It is now clear that the HIV group of retroviruses came from primates and that the origin of HIV1 is the chimpanzee subspecies of Central Africa, Pan troglodytes troglodytes. Many African primates are natural hosts of retroviruses and details of the natural history of both hosts and viruses are essential to understand the evolution of the latter. Data on the demography, ecology and behaviour of three species of primates (gorillas, chimpanzees and mandrills), studied in the Lope Reserve in Central Gabon since 1983, are analysed to identify the factors that allow, or favour, disease transmission within each species, between different species and between primates and humans. The comparison of the relative degree of risk suggests that of the three species, chimpanzees are the most susceptible to exposure to infection both from conspecifics and from other species. With respect to humans, the comparative analysis suggests greater exposure to viruses of mandrills and gorillas than to those of chimpanzees. For primates, major risk factors are: large social groups; bites inflicted in fights; social grooming; and predation on mammals. However, given that contacts between social groups of the same species are rare, the spread of a virus through a population will be slow and uncertain. Hunting wild animals is the behaviour most likely to provide transmission routes for primate viruses into human populations because of the high probability of blood-blood contact. Not only the hunters themselves, but also women who prepare bush meat for cooking and people involved in trade of carcasses are at high risk of transmission of pathogens. Hunting of bush meat is increasing in Central Africa due to the economic recession and the spread of logging into the forests of the interior of the region. To counter the significant risk of transmission of known, as well as new, diseases from primates to humans, urgent measures are needed to attack the root causes of commercial hunting which is not only risk to public health but also a serious threat to biodiversity in the region. PMID- 11030049 TI - [HIV-1 group N in Cameroon and apparent viruses in the chimpanzee]. AB - The hypothesis of the recent origin of AIDS by way of one of several events of inter-species transmission has been widely accepted. Whilst the primate HIV-2 reservoir has been clearly identified, the origin of HIV-2 is still uncertain. Since 1994, collaborative studies on the variability of HIV-1 conducted at the Yaounde Pasteur Centre have confirmed the very great diversity of circulating HIV 1 group M subtypes. This research has led to the identification of a further branch of HIV-1 phylogenesis, by the YBF30 strain, prototype for a new N group. A study conducted on chimpanzees isolated the retrovirus in three of them. Complete sequencing for one of the primates showed an important similarity in the env gene with the YBF30 strain, whereas the pol gene is closer to the HIV-1 group M. These findings support the hypothesis that HIV-1 group N resulted from a recombinant event. The origin of the HIV-1 group N pol fragment and more generally the origins of group M and O human viruses are still unclear. In order to complete the phylogenesis of human and non-human primate retrovirus as well as to identify two circumstances of more epidemiogenic variants, it is important to follow the variability of HIV-1 in central Africa and search for intermediary forms amongst simian species. PMID- 11030050 TI - [Epidemiology, origin and genetic diversity of HTLV-1 retrovirus and STLV-1 simian affiliated retrovirus]. AB - Human T Cell leukemia/lymphoma virus type I, the first human oncogenic retrovirus, is the aetiological factor of Adult T cell leukemia (ATL), a CD4+ malignant lymphoproliferative disease and of a chronic neuromyelopathy, the tropical spastic paraparesis or HTLV-1 associated myelopathy (TSP/HAM). HTLV-1, which infects from 15 to 25 million individuals world-wide, is highly endemic in certain areas such as south-western Japan, Central Africa, the Caribbean basin and some regions of South America, Melanesia and of the middle East (for example the Mashhad area of Iran). The three major modes of transmission for HTLV-1 infection are perinatal, sexual and by blood transfusion. Recent molecular studies on HTLV-1 have shown the existence of several molecular subtypes (genotypes). These are related to the geographical origin of the infected populations and not to the associated diseases. The virus has a very high genetic stability. Viral amplification via clonal expansion of infected cells, rather than by use of reverse transcription could explain this remarkable phenomenon which can be used as a molecular tool for gaining new insights into the origin, evolution and modes of dissemination of HTLV-1. Analyses of HTLV-1 and STLV-1 (the simian counterpart) viral strains from throughout the world suggest that four events are responsible for this pattern of dissemination: 1) the transmission in the wild of STLV-1 between simian species, 2) the transmission of STLV-1 to humans as exemplified by the high percentage of identity between STLV-1 strains from chimpanzees or from mandrills with some HTLV-1 strains present in inhabitants of Central Africa, 3) persistence of HTLV-1 over a long period of time (by sexual and perinatal transmissions) in remote populations, as seen in the Australo-Melanesian region and 4) a global distribution of HTLV-1 via large scale human migrations, e.g., the slave trade from Africa to the New World. PMID- 11030051 TI - [Forest ecosystems and Ebola virus]. AB - Despite data collected since the emergence of the Ebola virus in 1976, its natural transmission cycle and especially the nature of its reservoirs and means of transmission are still an enigma. This means that effective epidemiological surveillance and prevention are difficult to implement. The location of outbreak areas has suggested that the reservoir and the transmission cycle of the Ebola virus are closely linked to the rainforest ecosystem. The fact that outbreaks seldom occur suggests the presence of a rare animal reservoir having few contacts with man. Paradoxically, various serological investigations have shown a high prevalence in human beings, especially in forest areas of the Central African Republic (CAR), with no pathology associated. This would appear to suggest a circulation of both pathogenic and non-pathogenic strains as well as frequent contacts with man. The ecological changes resulting from human activity (agriculture and logging) account for the modification of the fauna (movement of rainforest fauna, introduction of savannah species) and could explain a multiplication of contacts. Likewise, it is interesting to note that the centre of outbreaks has always been in areas bordering on forests (ecotone foreset savannah in the Democratic Republic of Congo, savannah in Sudan). All these considerations have led us to establish a permanent "watch" in areas bordering on forests in the CAR, involving a multidisciplinary approach to the virological study (strain isolation, molecular biology) of the biodiversity of small terrestrial mammals. The results of a study conducted on 947 small mammals has shown for the first time the presence of the Ebola virus genome in two species of rodents and one species of shrew living in forest border areas. These animals must be considered as intermediary hosts and research should now focus on reservoirs in the ecosystem of forest border areas where contacts with man are likely to be more frequent. PMID- 11030052 TI - [Ecological basis for molecular surveillance of microbes and parasites transmitted by rodents]. AB - The multiplicity of circumstances by which a host can capture a microbiological agent can be broken down into two simplified processes: 1. encounter between host and microbe, 2. compatibility of two organisms. Given that rodents include species of generally small size, inhabiting continental areas in closer proximity to man than many other mammals, they offer numerous chain of contamination. Microbiological surveillance must both consider the emergence of old and new microbial types and identify ecological and demographic condition placing man at risk. Since it is materially impossible to survey all existing pathogens, priorities must be set according to empirical, epidemiological and ecological considerations. One of the major challenges is how to make microbiological surveillance more effective by way of new tools for detecting demographic changes in rodent populations. PMID- 11030053 TI - [Hantavirus: recent data and review of the literature]. AB - Hantavirus (Bunyaviridae family) are present worldwide and are the causative agent of haemorrhagic fever with renal or pulmonary syndrome. These viruses are transmitted by rodents with asymptomatic infection which thus act as an excellent reservoir. Molecular epidemiology indicates that Hantavirus may have evolved in three ways: mutations within the genome, reassortment of the segmented genome between two closely related Hantavirus, and genomic recombination, a relatively rare phenomenon among negative stranded RNA viruses. Each virus is carried primarily by a specific rodent host. This observation is confirmed by phylogenetic analyses: the evolution tree of the different Hantavirus, based on viral genomic sequences has been shown to mirror the evolution tree of their specific rodents, based on sequences of mitochondrial DNA. Altogether these data suggest that an ancestral Hantavirus infected a specific rodent, early during evolution, and was subsequently submitted to the same evolutionary pressure as the rodent host. PMID- 11030054 TI - [Genetic diversity of Lyssavirus]. AB - Rabies is a zoonosis of which the archaeological agents belong to the Lyssavirus species. Seven genotypes are known to exist. The isolates are grouped according to a) their geographical origin, b) historical context, c) type of vector. In parts of the world where several epidemic cycles exist, practical tools have been developed in order to distinguish between them more easily. Vaccine strains--all of which have been elaborated from genotype 1--offer little or no protection against the other genotypes. It is thus necessary to work on extending vaccine coverage. PMID- 11030055 TI - [Mycobacterium avium-intracellulare complex: phenotypic and genotypic markers and the molecular basis for interspecies transmission]. AB - The Mycobacterium avium complex (MAC) comprises a heterogeneous group of slowly growing mycobacteria that are pathogenic for both humans and animals. Two genetically distinct species within MAC are M. avium, which tends to infect HIV infected patients, and M. intracellulare more common among immunocompetent individuals. Contrary to M. intracellulare which relates to a single species, M. avium is separated into three subspecies; M. avium subsp. avium, a major opportunistic pathogen leading to a disseminated disease among terminal AIDS patients; M. avium subsp. paratuberculosis, causing Johne's disease among ruminants and implicated in Crohn's disease among humans; and M. avium subsp. silvaticum, a pathogen affecting birds that may cause chronic enteritis among calves but has not yet been associated with human disease. With the exception of mycobactin-dependent growth of M. paratuberculosis, most of the biochemical and cultural tests cannot discriminate among the three subspecies of M. avium. However, recently developed molecular methods and fingerprinting of strains using insertion sequences allows not only to distinguish among them but also further to explore the polymorphism of human and animal isolates. Numerous studies have underlined the probable role of various ecological niches (water, dust, soil, pigs, poultry and ruminants etc.) as a possible source of contamination for AIDS patients. This paper reviews the phenotypic and genotypic markers and epidemiology of M. avium complex organisms and current knowledge of the molecular basis of of inter-species transmission. PMID- 11030056 TI - [Intense campaign for vaccination with the oral polio vaccine: what are the repercussions on the enterovirus world?]. AB - To eradicate poliomyelitis and poliovirus, intensive vaccination campaigns with oral polio-vaccine (OPV) have been organised. Eradication campaigns may well be successful because the antiviral immunity and the local intestinal immunity due to OPV in particular avoids and/or limits poliovirus circulation. These campaigns give interesting opportunities for studying the impact of viral vaccines on the viral world in terms of ecological and genetic virology. The pre-eradication phase we are now entering brings with it two kinds of problems. First, the major disadvantage of OPV is the genetic and phenotypic variability of the vaccine strains. This variability leads to the spread of potentially pathogenic strains, which can be implicated in vaccine-associated paralytic poliomyelitis (VAPP). Genetic changes are characterised by point mutations and by genetic exchanges among OPV strains, between OPV and wild strains and perhaps between poliovirus and non-polio enteroviruses (ENPV). The fact that a few OPV mutant strains have been shown to multiply and/or to circulate for long periods suggests that OPV could sustain a reservoir of pathogenic poliovirus strains. Second, there are ecological considerations. The disappearance of wild poliovirus through OPV vaccination could be due not only to antiviral local immunity but also to competition between OPV strains and wild strains for infecting the digest tract. Moreover, a competition between OPV and other enteroviruses may take place in a common ecological niche. To our knowledge, the possible impact of intensive OPV vaccination campaigns on the ENPV populations has never been considered. Because the goal of poliovirus eradication may be reached in the near future, there is worry as to the possible evolution of ENPV towards highly epidemic and pathogenic strains. This is leading those laboratories involved in poliomyelitis surveillance not only to search for remaining wild poliovirus strains but also to study the possible long-term circulation of OPV strains and to develop efficient ENPV surveillance. PMID- 11030057 TI - [Circulation of the poliovirus in endemic zones with children vaccinated by the oral polio vaccine]. AB - Strategies aiming to eradicate the poliovirus and poliomyelitis seek primarily to eliminate wild strains associated with the disease, by means of world wide vaccination campaigns using the oral attenuated vaccine (OPV). OPV contains attenuated viral strains which retain their replicating capacity in the digestive tract and thus induce the development of an antiviral local intestinal immunity and limit the circulation of the virus. In such a context, poliomyelitis surveillance laboratories should study above all cases of acute flaccid paralysis (AFP), highlighting the circulation of wild strains, identifying regional reservoirs and guiding vaccination strategies. Alongside circulation, there appear to be important genetic and phenotypic shifts in vaccinating strains, since the OPV is capable of preserving a reservoir of pathogenic stains and thereby impairing vaccination efficacy and the eradication of the virus. Furthermore, non-polio enteroviruses should be considered as a source of emerging pathogenic strains. These questions are being studied by the Pasteur Institute with the objective of determining the effects of OPV campaigns on the circulation of the poliovirus. We have studied the poliovirus vaccine and the circulation of wild strains in urban and peripheral urban areas in African countries known to be endemic for poliomyelitis (Central African Republic, Madagascar, Cote d'Ivoire). The study population consisted of children who had already been vaccinated and new-borns in the course of vaccination. We also evaluated the diffusion of the vaccine strains in their immediate environment. Genetic interchanges were taken into account. For children who received the 3-4 OPV doses, asymptomatic virus excretion was insignificant (0.4-2.4%). The rate of virus excretion in the surrounding environment of children in the course of being vaccinated was relatively low (1.76-5.3%). Our study also detected variant and recombinant strains. PMID- 11030058 TI - [Genetic evolution under vaccine pressure: the Bordetella pertussis model]. AB - A possible genetic selective pressure related to the long-term use of vaccines has been the object of recent theoretical thought and publications. For more than thirty years, an effective vaccine has been in use against whooping cough on a wide scale basis in several countries. Thus, the Bordetella pertussis model may contribute to the analysis of an evolutionary risk linked to the vaccine. To maintain and improve the control of whooping cough, better vaccination coverage must be achieved in countries where prevalence is low. In countries where high vaccination coverage has been achieved over a long period, a trend toward the resurgence of the disease has been observed. Efforts are therefore now being directed toward primary vaccination and boosters. These two targets require new vaccines with fewer side effects. Outbreaks in highly vaccinated populations have been reported, raising the issues of vaccine efficacy, of the long-term effect of vaccines on the transmission of the disease, and of genetic selective pressure. Time trend modifications of circulating strains related to vaccination practices and vaccine types have been observed and are compatible with a selective pressure of the vaccine on related pathogens. However, evidence for a causal relation is lacking. In order to monitor and understand the various effects the vaccine may be having on the effectiveness of immunisation against whooping cough, further surveillance is needed, integrating a standardised characterisation of circulating strains and vaccines by way of a space-time sampling model. PMID- 11030059 TI - [Satellite data and disease transmission by vectors: the creation of maps for risk prediction]. AB - The complex set of criteria determining the reproduction rate of infection (distribution, density, dynamics of arthropod populations, temperature, humidity...) can be estimated with far greater precision by satellite than by conventional meteorological stations. By linking a) distribution modes and type of environment and b) satellite data and rates of underlying biological processes, we have been able to elaborate two super-imposed models: one for vector populations and one for pathogenic agents. We have used such models to look at, for example, trypanosomiasis, malaria and Lyme disease. PMID- 11030060 TI - [Malaria and schistosomiasis: 2 examples using systems of geographical information and teledetection in Madagascar]. AB - Schistosomiasis and malaria constitute major health problems in Madagascar. The main objectives of the national schistosomiasis control programme--launched in 1998--are to improve knowledge about the modes of transmission of the disease and conduct mass treatment in hyperendemic areas, so as to lower incidence rates. A Geographic Information System (GIS) was established aiming to conduct a series of remote sensing studies based on digital image processing and analysis from Landsat TM and panchromatic Spot. The importance of local environmental and geographic characteristics in the Ihosy region such as proximity to roads and water were assessed by spatial analysis in order to construct a predictive model of Schistosoma mansoni endemicity. Unstable transmission in the Central Highlands of Madagascar caused severe outbreaks of malaria in the 1980s. Attempts to prevent such events reoccurring have been highlighted in the national malaria control programme. Thanks to intense vector control measures introduced in the area over a five-year period, a marked decrease in incidence of malaria was observed. This region borders on the Highlands as well as southern areas, allowing for vector control evaluation. The GIS and remote sensing system were applied to analyse ricefield areas, where the main vector are Anopheles funestus. Rice-field surfaces were statistically linked with abundance of vectors evaluated by entomological parametres. PMID- 11030061 TI - [DNA chip technology]. AB - DNA chip technology has greatly evolved over the last decade and, associated with complete genome sequencing, is in the process of introducing a revolution into biological research. It is providing new and unique tools for studying emerging diseases outbreaks and epidemics. Nevertheless, microbiological surveillance, medical diagnosis, and field work involve a number of difficulties for which these new techniques have not yet been validated. Currently available chips are still limited in their application, but offer a powerful and economical alternative to former methods and will undoubtedly offer a range of unexplored applications in coming years. PMID- 11030063 TI - [Genotype and strains of Iranian Helicobacter pylori based on PCR-RFLP of conserved and non-conserved genes]. PMID- 11030062 TI - [Molecular epidemiology of large bacterial endemics in Sub-Saharan Africa]. AB - Over the past decades, the differentiation of bacterial strains for epidemiological purposes had been based on conventional phenotypic characters. More recently, methods studying the directly coded molecules or semantides (nucleic acids or proteins) have allowed, concomitantly with the technical progresses of electrophoresis, the description of stable, discriminant, reproducible markers, which were applicable to large series of isolates. Initially applied to study nosocomial infections in industrialised countries, these methods appear to be particularly suitable for an approach of the epidemiology of endemic bacterial infections in sub-Saharan Africa. The fact that these tools remain costly and technically complicated explains that most of these studies are conducted in the laboratories of industrialized countries. This research reveals the epidemiological complexity of most of these infections. Thus, the epidemiology of trachoma was studied by the analysis of polymorphism of the major outer membrane protein gene of Chlamydia trachomatis in a village of Gambia. A PCR based technique was used to determine the frequency of infection in symptomatic and clinically negative subjects and to specify the prevalence of the genotypes. The epidemiology of plague was studied by the restriction fragment length polymorphism (RFLP) analysis of the ribosomal RNA genes (ribotyping). Distinct ribotypes differentiated the strains of the first two pandemics from the third one. The strains of African origin were particularly heterogeneous, especially in Kenya. This diversity may be explained by the fact that the plague focus is extremely ancient in Central Africa. Bacterial agents of meningitis were also studied. The electrophoretic polymorphism of outer membrane proteins of Haemophilus influenzae of b type was used to specify the epidemiology of meningitis in Gambia. The invasive strains exhibited distinct profiles from non invasive strains. Different types were evidenced in the west, east and central parts of the country. The antigenic polymorphism of outer membrane proteins of Neisseria meningitidis allowed the differentiation of the strains isolated in Mali according to the period of isolation. Thus, the endemic strains of A serotype were distinguished from those belonging to the same serotype, which were responsible for the 1994 epidemic. Several molecular methods were applied to the typing of Vibrio cholerae strains, particularly those of the seventh pandemic. The enzyme electrophoretic polymorphism (MLEE), a technique based on RFLP analysis of toxin genes, the arbitrarily primed PCR (AP-PCR) and mainly the ribotyping were applied. This last method revealed that in Africa several clones of V. Cholerae El Tor were responsible for the seventh pandemic. Moreover the technique has evidenced the intercontinental spread of a clone of V. Cholerae isolated in 1993 in Calcutta and identified a year later in Guinea-Bissau. Tuberculosis is at present the first opportunistic infection linked to HIV infection in sub-Saharan Africa. Tuberculosis incidence is particularly high and is expected to increase. Several molecular methods, including IS 6110 RFLP analysis, AP-PCR and spoligotyping were used to study the epidemiology of tuberculosis in various countries: South Africa, Tanzania, Zimbabwe, Kenya and Malawi. The aims of this research varied: prevalence of reactivation and of recently acquired infections, routes of contamination, degree of genetic diversity of the organisms isolated in a given geographic area, urban and rural origins of the infections, comparison of isolates from HIV seropositive and HIV seronegative patients. Identical profiles in the strains isolated from several patients could correspond to clusters of infections. However, the identification of epidemiological links in most clusters is hard to obtain. (ABSTRACT TRUNCATED) PMID- 11030064 TI - [Prevalence of circulating endothelial cells (CEC) and expression profile in cerebral malaria]. PMID- 11030065 TI - What's new in neuroimmunology? PMID- 11030066 TI - Update on perinatal hypoxic insult: mechanism, diagnosis and interventions. AB - Cerebral hypoxia-ischaemia in the neonate can produce irreversible tissue injury and is always associated with major perturbations in the energy status of the brain. The major neurological manifestations of brain injury in these babies are spastic motor deficits. Different pathogenetic mechanisms may underlie hypoxic ischaemic injury of the brain such as decreased blood flow autoregulation, altered cerebral metabolism, thrombosis, haemorrhage, accumulation of toxic metabolites such as glutamate, impaired intracellular calcium turnover, release of interleukins and prostaglandins, iron accumulation and overproduction of free radicals. In summary, hypoxia-ischaemia in neonates triggers a cascade of biological processes culminating in cell death. Important advances in the assessment of cerebral injuries in neonates have been made in the area of neuroimaging, especially in magnetic resonance imaging which may provide useful prognostic information when obtained in the course of brain injury. Future studies may focus on new therapeutic pharmacological and non-pharmacological strategies aimed at the reversal of the pathophysiological mechanisms activated by perinatal hypoxia-ischaemia. PMID- 11030067 TI - Abnormalities in urinary pterin levels in Rett syndrome. AB - The pterins, neopterin and biopterin, occur naturally in body fluids including urine. Increased neopterin levels are associated with activation of the cellular immune system and reduced biopterins are essential for biosynthesis of the monoamine neurotransmitters. The present study measured urinary neopterin and biopterin by high-performance liquid chromatography in 40 subjects with Rett syndrome, eight of their healthy sisters and 29 female control volunteers (age range 2-54 years). The results confirm earlier preliminary findings that urinary neopterin levels are raised in a proportion of young girls with Rett syndrome but not in the older women. In contrast urinary biopterin levels are not different from controls in the youngest children but remain low while control values increase with age. These findings may indicate immune activation during the regression phase of Rett syndrome but also raise the possibility that an inherited fault in tetrahydrobiopterin metabolism increases the risk of developing the disorder. PMID- 11030068 TI - Delayed diagnosis of Duchenne muscular dystrophy. AB - A late diagnosis of Duchenne muscular dystrophy has implications for both child and family. This repeat audit has shown that the diagnosis continues to be delayed. The failure to recognize that non-motor, and specifically speech and language delay are common features of this disease may detract from the motor difficulties in affected children and contribute to the late diagnosis of this disorder. In the absence of a national newborn screening programme for Duchenne muscular dystrophy, all health care professionals should be made more aware of the condition and have a lower threshold for measuring a creatine phosphokinase level. PMID- 11030069 TI - Mild axonal neuropathy of children during treatment for acute lymphoblastic leukaemia. AB - Neurophysiological functioning was studied prospectively in children treated for acute lymphoblastic leukaemia with a low dose vincristine regime (8 x 1.5 mg/m2/dose), to obtain more insight into vincristine neuropathy. A WHO neurotoxicity score was estimated and vibration sense and electrophysiological measurements were taken at standardized times during vincristine treatment. The WHO neurotoxicity score showed decreased or disappearance of Achilles tendon reflexes, and mild sensory disturbances, but a grade 3-4 neurotoxicity was not demonstrated by any of the children. Vibration perception thresholds increased progressively during treatment and amplitudes of action potentials of peroneal and sensory ulnar and median nerves decreased, whereas nerve conduction velocities stayed unchanged. Both vibration perception thresholds and the electrophysiological findings hardly exceeded the limits of normality. We conclude that children treated for acute lymphoblastic leukaemia with a low dose vincristine regimen have mild axonal neuropathy which may be responsible for the motor problems in these children. PMID- 11030070 TI - A family with X-linked dominant Charcot-Marie-Tooth caused by a connexin32 mutation. AB - A family with a hereditary peripheral neuropathy is presented. Pedigree analysis suggested X-linked dominant mode of inheritance. The index patient became symptomatic at the age of 12 years. Clinical examination at 14 years revealed footdrop on the left, bilateral pes cavus, slight atrophy of thenar eminences, decreased muscle strength in both legs and brisk deep tendon reflexes. Electrophysiological studies were compatible with an axonal neuropathy. A novel point mutation located in codon 126 of the connexin32 gene, substituting a histidine for a tyrosine, was found in the index patient, in the mother, in two sisters and in a brother. The mother and the eldest sister had pes cavus bilaterally although they were asymptomatic. The younger brother and sister showed no signs of the disease. PMID- 11030071 TI - Peroneal nerve palsy: the role of early electromyography. AB - Footdrop is rare in the newborn period. In this report, the clinical and electrophysiological features of a newborn whose peroneal neuropathy was detected in the delivery room is presented. Her first electrophysiological examination, at the 13th hour of life, revealed a peroneal nerve lesion. All findings elicited in this and subsequent examinations suggested an antenatal onset. Clinical recovery began within weeks of the birth and she was able to walk, with no residual neurological abnormality, by her first birthday. Early electrophysiological examination of a newborn with peripheral neuropathy may provide valuable information about the time of onset and pathophysiological features of the nerve lesion. In spite of the early electromyogram findings revealing severe nerve injury, a good prognosis is possible in newborns with peroneal neuropathy. PMID- 11030072 TI - Visual loss as the presenting sign of Jeune syndrome. AB - We describe a 5-year-old girl with Jeune syndrome. The presenting symptoms included visual loss and night blindness only. In the neonatal period she had surgery on both hands for bilateral polydactyly. Following clinical and radiological examination a diagnosis of Jeune syndrome was proposed. Although Jeune syndrome is often characterized by multiple organ abnormalities, our patient presented with ocular symptoms at the age of 5 years. PMID- 11030073 TI - Recent developments in the epidemiology of Helicobacter pylori. AB - Prevalence determinations have been performed around the world, and regardless of how exotic a location, H. pylori is found in a substantial proportion of the population. H. pylori remains among the most universal of infections. Understanding of some features of infection has changed. Infection can be gained and lost at rates higher than previously realized. Oral-oral and oral-fecal transmission account for most, if not nearly all, cases of infection. H. pylori infection has declined rapidly in developed countries, which probably has contributed to declines in duodenal ulcer disease and gastric cancer. The full health implications of the potential elimination of infection are unknown. PMID- 11030074 TI - Gastritis and gastric cancer. Western countries. AB - Helicobacter pylori infection is the cause of chronic gastritis that progresses to atrophic gastritis over years and decades in more than half of affected individuals. H. pylori gastritis and, particularly, subsequent atrophic gastritis increase the risk for gastric cancer on multifactorial basis. Largely unknown cascades of manifold reactions result in gene errors of epithelial cells in gastric and atrophic stomach, which raise the likelihood of gastric neoplasias and cancer among people infected by H. pylori. The prevalences and incidences of gastric cancer and H. pylori are similarly decreased during the past decades in western countries, supporting the view that H. pylori infection is a key event and a trigger of the phenomena that result in cancer in some of the infected subjects. PMID- 11030075 TI - Malignant tumors of the stomach. Gastric mucosa-associated lymphoid tissue lymphoma and Helicobacter pylori. AB - With the help of many clinical studies, the diagnosis and therapy of gastric MALT lymphoma have evolved. Major progress has been seen in this area, including improvement of biopsy diagnosis, better histologic classification, new information concerning pathogenesis, and, especially, the start of a revolution in the treatment of low-grade gastric MALT lymphomas by eradicating H. pylori. About 12 clinical studies with almost 400 patients and case reports have shown that cure of H. pylori infection is associated with complete remission in approximately 80% of patients with low-grade MALT lymphoma in an early clinical stage. To establish H. pylori eradication as the primary choice in low-grade gastric MALT lymphoma further, it is necessary to select patients before therapy who are most likely to benefit from this single treatment modality. An excellent histologic workup of obtained biopsy specimens and comprehensive clinical staging are necessary. Because of the supposition that H. pylori-related growth support may play a role only in the early stages of low-grade gastric MALT lymphoma, the importance of determining the depth of lymphoma infiltration in the gastric wall is evident. Examinations by endosonographic ultrasonography have been shown to be the most reliable method to differentiate the layers of the gastric wall and to determine the infiltration depth of lymphomas. Eradication of H. pylori has to be considered as a first-line and single treatment modality in patients with low grade gastric MALT lymphoma in clinical stage EI1. As a therapy with fewer side effects than radiation, surgery, or chemotherapy and as a stomach-conserving treatment, eradication of H. pylori in patients with low-grade gastric MALT lymphoma should be the treatment of the choice within clinical trials because there are no long-term results available thus far. Besides pretreatment patient selection, careful follow-up with endoscopy, biopsies, and clinical staging including endoscopic ultrasonography is necessary. A 5- to 10-year follow-up is necessary before the definitive value of H. pylori eradication can be established, but long-term results are excellent thus far. There are many questions to be addressed: What are the exact mechanisms that lead to the malignant transformation of a reactive infiltrate? Why do approximately 20% of low-grade MALT lymphomas not regress after H. pylori eradication? Is there a molecular-genetic or immunologic point of no return? What is the biologic significance of the immunoglobulin rearrangement detected with PCR? What will be the 5- and 10-year relapse-free survival rates of patients suffering from low grade MALT lymphoma treated with H. pylori eradication alone as first and only treatment? The wave of new data each year about the role of H. pylori in gastric MALT lymphoma may help many of these questions to be answered. PMID- 11030076 TI - Gastritis and gastric cancer. Asia. AB - In Japan, high prevalences of H. pylori infection and AG are found, which seem to be causes for the high incidence of gastric cancer. H. pylori infection is not a sole cause of gastric cancer, however. Environmental factors, including a diet high in salt, also might be involved. For accurate diagnosis of H. pylori infection and AG, it is important to determine the locations from which the biopsy specimens are taken. Comparison of the severity of AG between different countries should be made using the same biopsy sampling system and grading criteria. Bayerdoerffer et al showed antral predominance of H. pylori colonization, but Genta et al reported more even distribution in the stomach. The authors' patients with NAG and those with antral and mild AG had similar scores of density at the six biopsy sites, whereas those with more extensive AG had the highest score at the middle body on the greater curvature. If only one biopsy specimen should be taken for the diagnosis of H. pylori infection in patients with extensive AG, the middle body of the greater curvature (site 6) would be the most suitable site. Western researchers reported that the grade of chronic and acute inflammation was significantly lower in the body than in the antrum. The authors' findings of patients with NAG were similar to those of Western researchers, but those with atrophy were not. Grades of inflammation and activity in the antrum decrease with the extension of AG. In the authors' patients, the earliest finding of the development of atrophy was found at site 1, and this was the most predominant site of atrophy in patients with extensive AG. Extension of AG into the body was determined by the finding of a biopsy specimen taken at site 3. The earliest finding of the development of intestinal metaplasia was found mostly at the middle body of the lesser curvature and next at the incisura angularis. In patients with extensive intestinal metaplasia, the scores of intestinal metaplasia at site 1 and site 3 were as high as the score at site 2. Site 6 was the last place for the invasion from the extension of AG. If biopsy specimens are taken from site 1, site 3, and site 6, they are enough for the assessment of the extent of AG and intestinal metaplasia. The Updated Sydney System recommended five biopsy sites including the incisura angularis, and its usefulness was coincidentally supported in Japan with a high prevalence of AG seen in results. PMID- 11030077 TI - Trend toward a reduced prevalence of Helicobacter pylori infection, chronic gastritis, and gastric cancer in Japan. AB - It is speculated that declines in H. pylori infection and gastritis over the past few decades may lead to a decline in gastric cancer in Japan, supplemented by excellent procedures for the early detection of gastric cancer. Because H. pylori infection rarely is acquired in adult life, once it is eradicated, reinfection would not be expected in adult patients. It seems likely that adequate treatment of H. pylori infection would provide long-term protection against gastric cancer. PMID- 11030078 TI - Bacteriology and taxonomy of Helicobacter pylori. AB - As the scientific community approaches the twentieth anniversary of the first isolation of H. pylori, it appears that despite the wealth of articles published in journals throughout the world every month, there are still many unanswered questions about the microbiology of this bacterium and others in the genus Helicobacter. PMID- 11030079 TI - Virulence and pathogenicity of Helicobacter pylori. AB - Bacterial and host response factors play significant roles in the pathogenicity of H. pylori-related disease manifestations. The complete DNA sequences for two H. pylori strain genomes have been published. The differences in the sequences between these two unrelated strains may enable clinicians to identify rapidly other conserved and potentially virulent genes and products. Whether these two DNA sequences are sufficient representation of the H. pylori genetic heterogeneity is unknown. The host immune response and the cascade of events that occurs with H. pylori infection are being clarified rapidly. Understanding the role of this gastric bacterium in apoptosis and cellular proliferation would enable clinicians to understand its relationship to ulcerogenesis and gastric malignancy. Piecing together many observations related to H. pylori would result in understanding the interaction of H. pylori factors and host responses that lead to the variety of disease manifestations associated with this chronic infection. The development of animal models with H. pylori and other Helicobacter species has set the stage in which in vitro observations can be tested in the in vivo model. PMID- 11030080 TI - Immunology of Helicobacter pylori and prospects for vaccine. AB - Since the initial discovery of H. pylori by Marshall and Warren 17 years ago, much progress has been made in treating this infection. However, as we enter the millennium, H. pylori infection continues to be one of the most common infections of mankind. In addition, eradication of H. pylori still requires multiple antimicrobial agents. A better understanding of the host immune response to H. pylori infection should allow investigators to develop immunotherapies to prevent the acquisition of infection and eradicate existing chronic H. pylori infection. PMID- 11030081 TI - Helicobacter pylori gastritis and gastric physiology. AB - It is now recognized that Helicobacter pylori infection exerts profound and diverse effects on gastric acid secretory function and that the alterations in acid secretion depend on the pattern of gastritis caused by the infection. In patients with an antral predominant nonatrophic gastritis, there is acid hypersecretion leading to duodenal ulcer disease. In patients with an atrophic pangastritis, there is markedly reduced acid secretion and increased risk for gastric cancer. It is now recognized that acid secretion also modifies H. pylori gastritis and a person's premorbid acid secretory status may be an important factor in determining the pattern of gastritis that an individual develops. This two-way interaction between H. pylori gastritis and gastric acid secretion is important in understanding the role of H. pylori infection in the response to proton-pump inhibitor therapy: It explains the more profound control of gastric acid secretion in H. pylori-positive patients and why rebound acid hypersecretion is confined to H. pylori-negative subjects. PMID- 11030082 TI - Gastric pathology associated with Helicobacter pylori. AB - A bacterium is associated with a specific gastritis. Neutrophils infiltrate the necks of the glands, just deep to the infected foveolae. This infiltration rarely, if ever, occurs without H. pylori infection. Foveolar epithelial damage is common, with loss of cell structure. Electron microscopy suggests that the bacteria cause this damage as they attach to the superficial cell membrane. These features, defined by Whitehead et al as active changes, appear specific for H. pylori infection. The neutrophils and specific epithelial changes disappear within days of starting treatment for Helicobacter. They rapidly recur if the treatment is unsuccessful. Without treatment, the changes remain for decades and are severe in 10% to 20% of cases. Other changes occur in the mucosa. Reduced mucus secretion occurs in damaged or proliferating epithelium. This reduced secretion occurs near healing ulcers or with other types of inflammation but is often severe when Helicobacter is present. It returns to normal within weeks of treating the infection. The bacteria adhering to the cell membrane may cause this change directly. Lymphoid infiltration occurs with any type of chronic inflammation or immune reaction. The infiltration is not specific for Helicobacter, and it reduces slowly in months or years after eradication of H. pylori. Peptic ulceration, particularly duodenal ulceration, although not specific, is particularly common with H. pylori infection. The long-term inflammation probably causes other gastric pathology. Atrophy is common. Epithelial metaplasia occurs in about 20% of patients, usually mild. Other features, such as scarring, epithelial dysplasia, and in situ malignant change, are less common. They show little improvement after eradicating H. pylori. The part played by the bacteria in their cause remains uncertain. Pathologists see a long-standing chronic gastritis clearly related to a bacterium. The inflammation often is severe and commonly damages the mucosa, with ulceration, atrophy, metaplasia, and occasional premalignant changes. Physicians would treat inflammation of this degree in most other parts of the body. This disease is usually symptomless. There is some controversy, but eradicating Helicobacter often fails to improve nonulcer dyspepsia. This failure results in a continuing argument over whether or not to treat the infection. Meanwhile the pathology continues. A temporary solution to the problem is suggested: Patients infected with Helicobacter can give informed consent. Patients can be told about the infection, the pathology, the poor relationship to symptoms, and side effects of therapy, and they can decide. PMID- 11030083 TI - N3-oxoacyl derivatives of L-2,3-diaminopropanoic acid and their peptides; novel inhibitors of glucosamine-6-phosphate synthase. AB - Novel inhibitors 1-4 of glucosamine-6-phosphate synthase from Candida albicans have been designed based on acylation of the N3 amino group of L-2,3 diaminopropanoic acid with the corresponding ketoacids. These inhibitors have been shown to alkylate the fungal enzyme in a time-dependent manner. Compound 3 containing trans-beta-benzoyl acrylic acid as an acyl residue was found to be the most potent inhibitor in the series. Dipeptides composed of the active inhibitors and norvaline demonstrated potent antifungal activity against selected strains of Candida spp. and Saccharomyces cerevisiae. Their activity was reversed upon addition of N-acetylglucosamine to the medium. PMID- 11030084 TI - Carbonic anhydrase inhibitors: inhibition of isozymes I, II and IV by sulfamide and sulfamic acid derivatives. AB - Sulfamide and sulfamic acid are the simplest compounds containing the SO2NH2 moiety, responsible for binding to the Zn(II) ion within carbonic anhydrase (CA, EC 4.2.1.1) active site, and thus acting as inhibitors of the many CA isozymes presently known. Here we describe two novel classes of CA inhibitors obtained by derivatizations of the lead molecules mentioned above. The new compounds, possessing the general formula RSO2NH-SO2X (X = OH, NH2), were obtained by reaction of sulfamide or sulfamic acid with alkyl/arylsulfonyl halides or arylsulfonyl isocyanates. A smaller series of derivatives has been obtained by reaction of aromatic aldehydes with sulfamide, leading to Schiff bases of the type ArCH = NSO2NH2. All the new compounds act as strong inhibitors of isozymes I, II and IV of carbonic anhydrase. Their mechanism of CA inhibition is also discussed based on electronic spectroscopic measurements on adducts with the Co(II)-substituted enzyme. These experiments led to the conclusion that the new inhibitors are directly coordinated (in a monodentate manner) to the metal ion within the enzyme active site, similarly to the classical inhibitors, the aromatic/heterocyclic sulfonamides. PMID- 11030085 TI - Glucosylated isoflavones as DNA topoisomerase II poisons. AB - Since topoisomerase poisons allow the enzyme to cut and covalently bind to DNA but abort the subsequent rejoining of the molecule after relieving the torsional stress. To study their action we have made use of a supercoiled form of the pRYG plasmid that bears a specific topoisomerase recognition and binding region. The conversion of the supercoiled circular double-stranded DNA to the linear and open circle forms in the presence of a topoisomerase II poison and a denaturation step by proteinase K-SDS is indicative of the efficiency of our test agents to stabilize the cleavable complex. Using this system, three glucosylated isoflavones (6'-methoxy-pseudobaptigenin-7-O-beta-glucoside, genistin, and daidzin) isolated from cytotoxic chloroform and ethyl acetate extracts of Retama sphaerocarpa Boissier, were found to have the ability to stabilize the cleavage complex human DNA topoisomerase II. PMID- 11030086 TI - Expression in yeast and purification of functional recombinant human poly(ADP ribose)polymerase (PARP). Comparative pharmacological profile with that of the rat enzyme. AB - Human poly(ADP-ribose)polymerase (PARP) was expressed in the yeast line JEL1 under the control of a GAL promoter. Proteins were extracted and human recombinant PARP purified to apparent homogeneity. The pharmacological profile of this human enzyme was characterised in terms of the effects of known inhibitors of PARP belonging to various chemical families and this was compared with that of the rat enzyme purified from rat testes, using the same purification protocol. The rat and the human enzymes appeared very similar in terms of their sensitivities to those selected inhibitors. PMID- 11030087 TI - Carbonic anhydrase activators: synthesis of high affinity isozymes I, II and IV activators, derivatives of 4-(arylsulfonylureido-amino acyl)ethyl-1H-imidazole. AB - Based on the X-ray crystallographic structure of the adduct of human carbonic anhydrase II (hCA II) with the weak activator histamine (Briganti, F., Mangani, S., Orioli, P., Scozzafava, A., Vernaglione, G. and Supuran, C.T. (1997) Biochemistry, 36, 10,384-10,392), a novel class of tight-binding CA activators was designed by using histamine (Hst) as lead molecule. Thus, N-1-tritylsulfenyl Hst was synthesized by reaction of Hst with tetrabromophthalic anhydride followed by protection of its imidazole moiety with tritylsulfenyl chloride. After hydrazinolysis, it afforded a key intermediate which was derivatized at the aliphatic amino group. Reaction of the key intermediate with 4 fluorophenylsulfonylureido amino acids (fpu-AA) or 2-toluenesulfonylureido amino acids (ots-AA) in the presence of carbodiimides, afforded after deprotection, a series of compounds with the general formula fpu/ots-AA-Hst (fpu = 4 FC6H4SO2NHCO; ots = 2-MeC6H4SO2NHCO). Some structurally related dipeptides with the general formula fpu/ots-AA1-AA2-Hst (AA, AA1 and AA2 represent amino acyl moieties), were also prepared, by a strategy similar to that used for the simple amino acyl compounds above. The new derivatives proved to be efficient in vitro activators of three CA isozymes. Best activity was shown against hCA I and bCA IV, for which some of the new compounds (such as the Lys, Arg, His or the dipeptide derivatives) showed affinities in the 2-12 nm range (h = human; b = bovine isozymes). hCA II was on the other hand somehow less prone to activation by the new derivatives, which possessed affinities around 30-60 nM for this isozyme. Ex vivo experiments showed some of the new activators to strongly enhance red cell CA activity (180-230%) after incubation with human erythrocytes. This new class of CA activators might lead to the development of drugs/diagnostic tools for the CA deficiency syndrome, a genetic disease of bone, brain and kidneys. PMID- 11030088 TI - Dye affinity labelling of yeast alcohol dehydrogenase. AB - The interaction of yeast alcohol dehydrogenase (ADH) with the reactive chlorotriazine dye Vilmafix Blue A-R (VBAR) was studied. VBAR was purified to homogeneity on lipophilic Sephadex LH-20 and characterised by reverse phase HPLC and analytical TLC. Incubation of ADH with purified VBAR at pH 8.0 and 37 degrees C resulted in a time-dependent inactivation of the enzyme. The observed rate of enzyme inactivation (kobs) exhibited a non-linear dependence on VBAR concentration from 22 to 106 nmol, with a maximum rate of inactivation (k3) of 0.134 min-1 and kD of 141.7 microM. The inhibition was irreversible and activity could not be recovered by gel-filtration chromatography. The inactivation of ADH by VBAR was competitively inhibited by the nucleotides NADH and NAD+. These results suggest that VBAR acts as an affinity label at the nucleotide binding site of yeast ADH. PMID- 11030090 TI - Competitive inhibition of Trypanosoma brucei phosphoglucose isomerase by D arabinose-5-phosphate derivatives. AB - We report four new strong high energy intermediate analog competitive inhibitors of fructose-6-phosphate isomerization catalyzed by purified Trypanosoma brucei phosphoglucose isomerase: D-arabinonhydroxamic acid-5-phosphate, D-arabinonate-5 phosphate, D-arabinonamide-5-phosphate and D-arabinonhydrazide-5-phosphate. For comparison, the inhibitory properties of the corresponding non-phosphorylated analogues D-arabinonhydroxamic acid, D-arabinonate, D-arabinonamide and D arabinonhydrazide were also evaluated. D-Arabinonhydroxamic acid-5-phosphate appears as the most potent competitive inhibitor ever evaluated on a phosphoglucose isomerase with an inhibition constant value of 50 nM and a Michaelis constant over inhibition constant ratio of about 2000. Our results show that anionic high energy intermediate analogues, and more particularly D arabinonhydroxamic acid-5-phosphate, display a weak but significant specificity for Trypanosoma brucei phosphoglucose isomerase versus yeast phosphoglucose isomerase, while neutral high energy intermediate analogues are not selective at all. This would indicate the presence of more positively charged residues in the active site for Trypanosoma brucei phosphoglucose isomerase as compared to that of yeast phosphoglucose isomerase. PMID- 11030089 TI - Nickel-induced substrate inhibition of bovine liver glutamate dehydrogenase. AB - The effects of nickel ions on reductive amination and oxidative deamination activities of bovine liver glutamate dehydrogenase (GDH) were examined kinetically by UV spectroscopy, at 27 degrees C, using 50 mM Tris, pH 7.8, containing 0.1 M NaCl. Kinetic analysis of the data obtained by varying NADH concentration indicated strong inhibition, presumably due to binding of the coenzyme to the regulatory site. In contrast, almost no inhibition was observed in the forward reaction. The fact that nickel ions have the capacity to enhance binding of NADH to the enzyme was confirmed by an electrochemical method using a modified glassy carbon electrode. Use of NADPH instead of NADH showed only a weak substrate inhibition, presumably related to lower affinity of NADPH for binding to the regulatory site. Lineweaver-Burk plots with respect to alpha-ketoglutarate and ammonium ions indicated substrate and competitive inhibition patterns in the presence of nickel ions, respectively. ADP at 0.2 mM concentration protected inhibition caused by nickel. These observations are explained in terms of formation of a nickel-NADH complex with a higher affinity for binding to the regulatory site in GDH, as compared with the situation where nickel is not present. Such effects may be important for regulation of GDH and other NADH utilizing enzymes. PMID- 11030091 TI - [Water jet cutting for bones and bone cement--parameter study of possibilities and limits of a new method]. AB - Water jet techniques have been used in industrial cutting, drilling and cleaning applications for more than 30 years. Plain water is typically used for the cutting of non-metallic materials. The addition of abrasive substances to the stream allows almost any material to be cut. The first medical applications were reported in the early 1980s, when the water jet was used to cut organs. The present study investigates the use of water jet cutting technology for endoprosthesis revision surgery. Bone and PMMA (polymethylmethacrylate) samples were cut at different pressures using an industrial water jet cutting device. Using plain water at 400 bar, PMMA was cut selectively without damaging the bone; above 400 bar, bone was also cut, but the cutting depths in PMMA were significantly greater (p < 0.05). Adding a water-soluble abrasive disaccharide to the water results in a significantly higher removal rate for both materials (p < 0.05), but selectivity is lost, although the differences in cutting depth between the two materials was significant (p < 0.05). With an abrasive, the quality of the cut was better for both materials. The water jet technology--in particular the abrasive technique--can be used to cut biomaterials such as bone and bone cement. The diameter of the jet is a great advantage when working in the confined area at the prosthesis interface. The cutting process is essentially cold, thus eliminating a thermal effect, and the jet reaction forces are relatively low. Accurate manipulation of the hydro jet nozzle is possible both manually and by robot. The results obtained show that it is possible to remove prostheses with this cutting technique, rapidly and with little damage to the surrounding tissue. Problem areas are the development of sterile pumps and the "depth control" of the jet. PMID- 11030092 TI - [Precision and comparison of CT-, MRI- and DL-controlled interventions exemplified by lumbar facet infiltration--an experimental study]. AB - We evaluated the accuracy of the needle tip representation by different imaging techniques for the guidance of facet infiltrations. For visualisation of the lumbar facet joints we used a high-field magnetic resonance tomograph (MRT) with a 2.0 Tesla field and 3.5 mm slice thickness, an open low-field magnetic resonance tomography (MRT) with an 0.064 Tesla field and 9 mm slice thickness, and IMATRON electron beam computed tomograph (EBCT) with a slice thickness of 6 mm, and a mobile C-arm fluoroscope. The study was performed on 4 human cadaveric lumber spine preparations, each of which had 8 facet joints. Under imaging control, special injection needles were placed as close as possible to the facet joint space. Following placement of he needle, all specimens were scanned with the electron beam tomograph using a slice thickness of 1.5 mm. The thin-slice study served as the gold standard. The distance between the tip of the needle and the facet joint was measured in all the images. Comparison of the different modalities with the gold standard revealed the following results: 1) median values of the absolute differences were 1.25 mm for high-field MRI, 1.35 mm for 6 mm EBCT, 2.05 mm for low-field MRI, and 2.30 mm for X-ray fluoroscopy. 2) While there was no statistically significant difference in the accuracy of tip localization between high-field MRI and 6" EBCT (p = 0.293), both systems were more precise than low-field MRI (p = 0.04) and X-ray fluoroscopy (p = 0.009). When choosing the best imaging technique, such additional factors as radiation, costs and time, must also be considered. Provided necessary radiological precautions are taken, and assuming careful pre-interventional planning, CT. EBCT and X-ray fluoroscopy are currently more effective than the expensive, time consuming and costly magnetic resonance tomography. PMID- 11030093 TI - [Cytotoxicity study of high gold content Degutan surfaces of various degrees of roughness with fibroblasts (BALB 3T3) and osteoblasts (hFOB 1.19)]. AB - The cytotoxicity of Degutan surfaces with different degrees of roughness, and the effect of surface structures on osteoblast proliferation and differentiation, was investigated with standardised cell culture systems. Fibroblast cell lines (BALB/3T3) and osteoblast cell lines (hFOB 1.19) were used. The number and variability of the cells were determined for assessment of proliferation and alkaline phosphatase activity, collagen I and osteocalcin production were used as parameters for differentiation. In the early phase, the largest numbers of cells and greatest proliferation were measured on polished Degutan surfaces. In the late phase, however, larger numbers of cells and a greater degree of proliferation were to be seen on sandblasted and sandblasted/heat-treated Degutan surfaces. No differences were found for collagen I, osteocalcin production or alkaline phosphatase activity. Neither the osteoblasts nor the fibroblasts revealed a toxic effect of Degutan. The results for osteoblast differentiation correlate with recent studies on identical structured titanium surfaces. In view of the immeasurable amount of ion release, Degutan may be considered an ideal model for an inert material surface. PMID- 11030094 TI - Temporal properties of high frequency intra-QRS signals in myocardial infarction and healthy hearts. AB - The prevalence of late potentials after myocardial infarction depends on the site of the infarction. This may be caused by the different activation onsets of the anterior and inferior myocardial segments. Therefore, in anterior infarcts the high frequency signals may be concealed within the QRS whereas in the inferior infarcts they last beyond the end of the QRS. We compared the timing and the spatial patterns of high frequency intra-QRS signals (IQSs) in the different infarction sites. We investigated 14 patients with anterior infarcts, 17 patients with inferior infarcts, and 10 healthy subjects. 31-lead magnetocardiograms were recorded in left precordial position and averaged. The QRS signals were smoothed with a Savitzky-Golay filter. The smoothed QRS signals were subtracted from the measured ones. The difference of the signals (frequency band of about 60-200 Hz) representing the high frequency components was quantified. The percentage of the high frequency signals was calculated for the entire QRS, for the first and for the second half, respectively. We found that in patients with anterior infarcts the high frequency components predominantly appeared in the first half of the QRS whereas in inferior infarcts these components predominantly appeared in the second half of the QRS. The different infarction sites were associated with different spatial patterns of the high frequency signals on the body surface. In healthy subjects there was not such a preferential association of time intervals and high frequency signals. Late potentials are the special case of high frequency signals appearing in the terminal QRS. It is the general property of the myocardium to generate high frequency signals associated with the depolarization of infarcted tissue. The timing of such signals and the spatial distribution patterns on the body surface may help to identify the location of the sources. PMID- 11030095 TI - [Expert system for aiding diagnosis in hearing tests]. AB - For expert systems intended to aid diagnosis, a structure with five levels is proposed. These levels are the original area, the parameter and a reduced parameter layer, the classification and the final-decision layer. On the basis of this structures, an expert system was developed specifically for neonatal hearing screening with transitory evoked otoacoustic emissions (TEOAE). In a second step, this system was investigated for its suitability to classify emissions, regardless of patient age. For the comparison measurements in 252 mainly adult patients, some with an acquired hearing impairment, were used. To adapt the pass/fail decision to the extended evaluation criteria, the false classifications from a first run with the new data were used for training. Thereafter, the expert system, working with a wider data basis, classified the new data with a sensitivity that was increased by 4.8% to 97.2%, and a 2.0% improvement in specificity to 95.5% when classifying new data, These results, together with those of 97.3% and 94.3% achieved with exclusively neonatal TEOAE classification, clearly show the advantage of the expert system structures chosen, and document evidence of the practical applicability of the method. PMID- 11030096 TI - [Significance of social-hygienic monitoring in the control of environmental quality and population's health]. AB - The paper deals with control of the population's environment and health. It shows the leading role of public health services in this activity. A standard approach to controlling environmental quality is presented. The first stage of this process is to determine the population's health status. To estimate the hazard of adverse environmental factors to human health may be a token of success in implementing health-promoting programmes. PMID- 11030097 TI - [Role of risk factors in the development of chronic digestive system diseases in children]. AB - The spread, patterns, and risk factors of alimentary diseases were examined in children residing in areas of petroleum-refining, petrochemical, and chemical industries. A total of 4816 children aged 3 to 14 years who lived in Ufa areas differing in the level and nature of ambient air pollution were examined. Those living in the poorer ecological areas (high total pollution and levels of chlorinated hydrocarbons, formaldehyde, and phenol) had more commonly gastroenterological diseases with prevailing hepatobiliary disorders in the pattern of diseases. The specific features of the diseases in these children are earlier onset, more frequent recurrences, more prolonged exacerbations involving other organs and systems than in those living in more favourable areas. PMID- 11030098 TI - [Effects of air pollution on stillbirth rates in an industrial town]. AB - The impact of seasonal variations in ambient air pollution with some compounds (suspended matters, sulphur dioxide, carbon oxide, nitrogen dioxide) on stillbirth was studied. Changes in the concentration of the studied pollutants were found to affect stillbirth. PMID- 11030099 TI - [Hygienic evaluation of sewage from coal open pits]. AB - The composition of open-pit, drainage, process, domestic sewages was studied in the stripping. They showed varying contents of substances and chemicals. The stripping sewage cannot be used for processing and agricultural purposes with pretreatment. A technique is proposed for local purification of open-pit and drainage waters before their mixing in the accumulator. PMID- 11030100 TI - [Ranging the territory of Serpukhov by the degree of soil pollution by chlorinated biphenyls]. PMID- 11030101 TI - [Impact of extreme climatic conditions on morphophysiological parameters]. AB - The paper shows that there are uniform changes in the somatotypical characteristics with increases in the age of able-bodied population and in their adaptation to emergency environmental conditions. The amount of an indefinite somatotype increased in all groups (by age, adaptation to the conditions, low- and high lands and high-latitude Antarctic region), major signs of ethnicity and regional traits of the populations remaining in these periods. PMID- 11030102 TI - [Antimicrobial lacquer Intercide is a novel promising formulation of disinfectants]. PMID- 11030103 TI - [The repellent and insecticidal effects of rustomectin against German cockroaches]. PMID- 11030104 TI - [Pathogenesis of tolylene diisocyanate intoxication]. AB - Experimental and clinical investigations to evaluate immunity, lipid peroxidation (LPO), and antioxidative defense (AOD) under the action of tolylene diisocyanate at the level of MAC and 10 MAC were conducted for further studies of the agent's pathogenetic mechanisms. The experimental investigations showed the predominant activation of cellular and humoral immunities, as well as the suppression of nonspecific defence at 10 MAC, enhanced LPO, and phasic changes in AOD. Most profound immunological changes were observed in those having as long as 5 years of length of service. PMID- 11030105 TI - [Specific features of adverse effects in workers exposed to different levels of methylmethacrylate]. AB - The relation of the incidence of malignant neoplasms to the dose of methyl methacrylate (MMA) was studied in 3588 workers. There was a clear correlation between the dose of MMA and the risk for cancer of the genital system in females and of the lung in males. Examining the immunity in groups receiving different doses of MMA revealed significant immunological disorders when exposed to its high concentrations. PMID- 11030106 TI - [Problems of nutrition and ensuring sanitary-epidemiological well-being in the Orlov region]. AB - Based on the data on hygienic monitoring of the actual food consumption in the population of the Oryol region and on the quality of foodstuffs, the authors developed hygienic recommendations whose introduction caused a great sociomedical effect: lower incidence of salmonellosis, dysentery, hepatitis virus A, and acute intestinal infections of unclear etiology. PMID- 11030107 TI - [Selenium and its role in nutrition]. AB - The review presents data available in the literature on the spread of selenium deficiency. Epidemiological surveys show that there is a potential risk for selenium deficiency in pregnant females and in 7-10-year-old children. The negative impact of radiation on serum selenium levels was evidenced by a survey of Chernobyl Atomic Power Station workers. Higher selenium levels in the hairs of the pubis, beard, celiac plexus, and axillary spaces than those in the hair of the head (by 1.41, 1.34, 1.36, and 1.14 times, respectively) confirm that there is a relationship between selenium and sexual hormones in the body. Dietary fiber concentrates that are derived from plant materials and contain a complex of structural carbohydrates and lignin may be used in combination with selenium compounds in human diets to prevent selenium deficiency. PMID- 11030108 TI - [Problems of vocational training and occupational hygiene in adolescents]. PMID- 11030109 TI - [An automated expert system for medical professional orientation of teenagers and youth]. AB - The paper outlines a concept of the automatic medical examination system (MES) designed to aid a physician during his/her medical professional orientation. The authors propose a general MES schedule, define the term "professional inability" (in the medical sense). They provide evidence for that it is necessary to use the assessments made experts as to the professional disability of adolescents and young people who have chronic diseases and dysfunctions. The common principles in the construction of expert MES matrices are outlined. After its completion, MES may be used for purposes of medical professional orientation. PMID- 11030110 TI - [Functional capacities of the body and their significance in the assessment of adolescents' health]. AB - Evidence is provided for the necessity of developing criteria for early detection of disorders in adolescence. Based on the results of comprehensive medical examination of 1940 teenagers, the authors have developed standards for an adaptative index. Differences have been found at functional maturity in male and female teenagers. There is an association of adaptive capacities with some diseases and the specific features of physical development in adolescents. It is concluded that the adaptative capacities may be used as a criterion for evaluating the health status. PMID- 11030111 TI - [Children's health promotion in the logopedic groups of preschool institutions]. AB - The specific features of the health status and development in preschool children who have marked speech disorders make it necessary to use a special physical educational system in combination with health-promoting measures to rehabilitate them at a preschool educational establishment. Evidence is provided for the efficiency of the system proposed. PMID- 11030112 TI - [Problems of iodine deficiency prevention under present-day conditions]. PMID- 11030113 TI - [State of the visual organ in pupils of a secondary school and a general education school]. AB - The impact of school loads on visual function was studied in 845 pupils from two general educational schools and two lyceums of Izhevsk. The studies confirmed the negative impact of school teaching on the children's vision. Negative trends were most noticeable in junior (1st-5th-form), girls, and children having additional classes, in the lyceums in particular. PMID- 11030114 TI - [Functional development of children's home inmates]. PMID- 11030115 TI - [Significance of morphologic parameters in hygienic studies]. AB - The paper demonstrates the potentialities of structural and functional analyses and some morphological parameters used in experimental and field studies. It identifies promising trends of morphological studies and their priorities and shows the need of introducing assessments of hepatic polyploidization into guidelines on the study of the overall toxic and embryotoxic action of the study agents. An algorithm has been developed to evaluate the mucosa during epidemiological surveys. PMID- 11030116 TI - [Toxicity and hazards of the herbicide starane 200]. AB - The paper deals with the burning problem in the evaluation of the toxicity and hazards of the new herbicide Starane 200. The main disadvantage of its use is ambient air pollution in the working area. The pollution preserves within 2 hours after its treatment. Starane 200 preserves in the soil and water within 60 and 10 days, respectively. The agent is classified as a moderately hazardous agent by the parameters of acute intragastric toxicity. The cumulative properties of the agent are slight. The ineffective dose of Starane 200 was 1.2 mg/kg given intragastrically to albino rats in a 12-month experiment. PMID- 11030117 TI - [Comparative evaluation of the effects of chromium and molybdenum on the digestive function]. AB - The paper describes chromium- and molybdenum-induced digestive functional changes and shows that there is a relationship of the presence, nature, and the degree of manifestation of their effect to the type of a metal, its dose, and the route of administration. PMID- 11030118 TI - [Effectiveness of disinfecting measures of epidemic control in Lipetsk]. AB - The paper characterizes the sanitary and epidemiological situation in Lipetsk in 1997-1998. It emphasizes the topical problems in the control of tuberculosis and particularly life-threatening infections at the present stage and the importance of prevention of nosocomial diseases. The paper also shows how actions in final disinfection are made in the foci of infectious diseases in the Lipetsk Regional Center of Disinfection and Sterilization. PMID- 11030119 TI - [Comparative study of the effectiveness of several insecticides in the control of house dust mites]. PMID- 11030120 TI - [Improvement of analytical surveillance of environmental water objects]. PMID- 11030121 TI - [Certification criteria for assessing whether the quality of drinking water meets the SanPiN standards 2.1.45.559]. PMID- 11030122 TI - [Determination of low concentrations of chloroform and 1,2-dichloroethane in the urine]. PMID- 11030123 TI - [Regression analysis in establishing relationship between population health and the environment]. PMID- 11030124 TI - Organ malperfusion in acute aortic dissection. AB - OBJECTIVE: Organ malperfusion in aortic dissection can precipitate a serious condition. The strategy of treatment for it has been controversial. We have focused on the strategy and outcome of acute aortic dissection with organ malperfusion. SUBJECTS AND METHODS: Between January 1995 and December 1998, 134 acute aortic dissection patients were admitted. There were 73 males (65.4 +/- 8.0 years old) and 61 females (66.7 +/- 7.4 years old). There were 83 patients of Stanford type A, and 51 patients of type B. Of them, 24 patients (17.9%) were complicated by organ malperfusion. The brain was affected in 4, the heart in 5, the spinal cord in 2, the liver in 1, the intestine in 1, the kidney in 4, and the lower extremities in 10 patients. Our management strategy for a patient with malperfusion in acute aortic dissection was that the antecedent operation was initially mandatory, and central grafting was secondarily considered. RESULTS: Refusal of operation or lethal conditions excluded 8 of the 24 patients from operation. An antecedent operation was mandatory in eight of the remaining 16 patients. The overall mortality was 33.3% (8/24 patients), and operative mortality was 31.3% (5/16 patients) in the patients with malperfusion. The overall mortality was 11.8% (13/110 patients), and the operative mortality was 11.1% (9/81 patients) in the patients without malperfusion. CONCLUSION: Organ malperfusion is a major component in the management and treatment of acute aortic dissection. Only an appropriate strategy and therapy could result in a satisfactory outcome. PMID- 11030125 TI - Strategy for reduction of stroke incidence in coronary bypass patients with cerebral lesions. Early results and mid-term morbidity using pulsatile perfusion. AB - OBJECTIVES: Cerebral complication is an important factor affecting the outcome after coronary artery bypass surgery under cardiopulmonary bypass. One of the causes for cerebral complication is preoperative cerebrovascular stenotic lesion. Here, we have studied the effect of pulsatile perfusion on the rate of cerebral complication due to a cerebrovascular lesion in patients undergoing coronary arterial bypass graft under cardiopulmonary bypass. METHODS: 261 consecutive elective patients underwent operation using cardiopulmonary bypass for management of the atherosclerotic ascending aorta. Group 1 consisted of 62 patients with a cerebrovascular stenotic lesion (> or = 75%) identified on a magnetic resonance angiogram or multiple cerebral infarction diagnosed using a computer tomogram. Group 2 consisted of 199 patients diagnosed with no significant cerebral lesion. In Group 1, the systolic blood pressure during cardiopulmonary bypass was maintained at a level of 80 mmHg by means of pulsatile flow. In Group 2, non pulsatile perfusion was used as usual. RESULTS: The overall hospital mortality was 1.5%, and no mortality was caused by a cerebral event. Only one patient in Group 1 suffered from temporary hemiparalysis. A cerebral complication occurred in only 1.6% in Group 1, and 0.4% overall. The actuarial freedom from cerebrovascular accident after 54 months was 84.4% in Group 1, and 96.2% in Group 2 (p = 0.0011). CONCLUSIONS: Management of the atherosclerotic ascending aorta and the use of pulsatile perfusion were helpful in preventing cerebral injury during CABG. PMID- 11030126 TI - Surgical management of aortopulmonary window. AB - OBJECTIVE: Aortopulmonary window is a rare congenital malformation involving a window-like communication between the ascending aorta and the pulmonary artery. Here, we present our experience regarding the surgical repair of an aortopulmonary window, and also assess the long-term outcome. METHODS: Thirteen children with an aortopulmonary window associated with various congenital lesions underwent a repair of the defect. The age at operation ranged from 3 days to 1 year (median age, 19 days). The patient's weight ranged from 2.1 to 7.0 kg (mean weight, 3.6 kg). The associated lesions included an interrupted aortic arch (5 patients), a ventricular septal defect (2), an atrial septal defect (1), mitral valve regurgitation (1), and tricuspid atresia [Ic] with mitral valve regurgitation (1). The aortopulmonary window was repaired with a cardiopulmonary bypass in 11 patients, and 2 patients were ligated without a cardiopulmonary bypass. RESULTS: One patient associated with tricuspid atresia died (mortality rate of 7.7%). There has been no late death during a mean follow-up of 7 years and 3 months. CONCLUSIONS: The surgical results for an aortopulmonary window are encouraging, even if such patients are associated with major cardiac anomalies and an interrupted aortic arch. Most have shown a good long-term outcome. PMID- 11030127 TI - Development and clinical application of minimally invasive cardiac surgery using percutaneous cardiopulmonary support. AB - OBJECTIVES: Optimal cardiopulmonary support during minimally invasive cardiac surgery remains controversial. We developed cardiopulmonary bypass for minimally invasive cardiac surgery using percutaneous peripheral cannulation. METHODS: Subjects were 34 patients (age: 58 +/- 13 years; range: 17-73) undergoing minimally invasive cardiac surgery using percutaneous cardiopulmonary support between June 1997 and March 1999. Procedures included atrial septal defect closure (n = 14), partial atrioventricular septal defect closure (n = 1), mitral valve replacement (n = 8), mitral valve repair (n = 3), aortic valve replacement (n = 6), coronary artery bypass grafting (n = 1), and right atrial myxoma extirpation (n = 1). Bicaval venous drainage from the right internal jugular vein and the femoral vein and arterial return to the femoral artery were instituted by percutaneous cannulation. Venous drainage was implemented by negative pressure ( 20 to -40 mmHg) and arterial return was by conventional roller pump. All procedures were conducted through a skin incision 8 +/- 1 cm, from 6 to 10 cm and partial sternotomy. Aortic cross clamping and cardioplegic solution were administered in the surgical field. RESULTS: The operation lasted 224 +/- 45 min., cardiopulmonary bypass 104 +/- 32 min., and aortic clamping 77 +/- 23 min.. No deaths occurred. One patient with residual atrial septal defect required reoperation through the same skin incision. Only 1 patient required homologous blood transfusion. The average postoperative hospital stay was 15 +/- 5 days. CONCLUSIONS: Minimally invasive cardiac surgery using percutaneous cardiopulmonary support is safe and an excellent option for selected patients affected by single valve lesion, simple cardiac anomalies, and coronary artery bypass grafting. PMID- 11030128 TI - Early surgery for active infective endocarditis. AB - OBJECTIVE: The timing of surgery for active infective endocarditis remains controversial. In this report, we have reviewed 26 patients who underwent surgery for active infective native-valve endocarditis between April 1992 and December 1998. PATIENTS AND METHOD: There were 19 male and 7 female patients (mean age 45 years). The aortic valve was involved in 8 patients, the mitral valve in 6 patients, tricuspid valve in 2 patients, both aortic and mitral valves in 7 patients, both aortic and tricuspid valve in 2 patients, and both mitral and tricuspid valve in one patient. The most common microorganisms were streptococcal species. Preoperative high New York Heart Association functional class (III and IV) was presented in 20 patients (77%). Progressive heart failure and the echocardiographic findings of vegetation (larger than 1 cm) were the main operative indications. Emergency or urgent surgery was required in 18 patients (70%). All patients underwent valve replacement, involving 25 mechanical prosthesis and 8 bioprosthesis. RESULTS: The operative mortality was 7.8% (n = 2). In the two patients who died, the infection had extended to the deep cardiac tissue and to the cerebral artery. The mean follow-up of the 24 survivors was 33 months (range from 6 to 82 months). There was no late death and no recurrence of infective endocarditis. CONCLUSION: In case of active infective endocarditis, early surgical intervention is recommended in patients with rapidly progressive cardiac deterioration or vegetation seen on echocardiography. PMID- 11030129 TI - Bypass graft material and myocardial protective procedure in combined coronary artery bypass grafting and valve surgery. AB - OBJECTIVE: The aortic cross clamping time is prone to be longer when coronary artery bypass grafting (CABG) is combined with valve surgery. Therefore, the myocardium that is revascularized by in-situ internal thoracic artery graft is at risk to ischemia, and, myocardial protection is especially important in such operation. In this study, the effect of myocardial preservation of combined antegrade, retrograde and terminal warm blood cardioplegia during combined valve surgery and CABG using the internal thoracic artery as a bypass conduit was evaluated. METHODS: From November 1992 to August 1999, 15 patients received combined CABG and valve surgery. Among these 15 patients, 13 patients who did not need hemodialysis were divided into 2 groups, and a comparative study was done. In Group I (n = 5), only the saphenous vein graft was employed for combined CABG and valve surgery, and myocardial protection was done by combined antegrade and terminal warm blood cardioplegia. In Group II (n = 8), at least 1 in-situ internal thoracic artery graft was employed for CABG and valve surgery, and myocardial protection was done by combined antegrade, retrograde and terminal warm blood cardioplegia. RESULTS: Despite longer aortic cross clamping time in Group II, the peak creatine kinase-MB of Group II was significantly lower. In addition, the postoperative administration of dopamine tended to be less in Group II. CONCLUSION: Myocardial protection by combined antegrade, retrograde and terminal warm blood cardioplegia may be an effective adjunct to combined valve surgery and CABG employing the in-situ internal thoracic artery graft. PMID- 11030130 TI - Traumatic cardiac rupture with acute ascites. AB - A rare accumulation of serous ascites following traumatic cardiac rupture is reported for a 21-year-old man transferred to the emergency center of our hospital from a community hospital suffering from severe shock due to a motorcycle accident. Computed tomography scan showed moderate pericardial and intrapelvic fluid accumulation strongly suggesting cardiac and visceral injuries. An emergency sternotomy disclosed a rupture of the right atrial appendage, successfully closed with primary sutures. A laparotomy was done to aspirate moderate serous fluid, which was clear and not bloody. Edema of the retroperitoneal space and hepatic congestion were noted with no accompanying organ injury. Ascites pathogenesis is unknown but appeared to be related to portal venous congestion induced by cardiac tamponade combined with massive intravenous fluid infusion done to correct the patient's deteriorating hemodynamics. PMID- 11030131 TI - Total anomalous pulmonary venous connection with bronchogenic cyst in neonatal period. AB - Here, we report a case of a two-day-old neonate with total anomalous pulmonary venous connection to the innominate vein and a bronchogenic cyst arising from the trachea. Antenatal echocardiography had delineated both cardiac and extracardiac lesions, and a repeated examination on the day of birth disclosed progressive enlargement in the cyst in a manner so as to obstruct the innominate vein. On the second day of life, the patient underwent complete correction of the cardiac lesion and total excision of the cyst. The patient recovered uneventfully and was discharged on the thirteenth postoperative day. PMID- 11030133 TI - Partial left ventriculectomy in a 3-year-old boy with dilated cardiomyopathy. AB - A 3-year-old boy suffered severe heart failure 2 months after ventricular septal defect repair. The cardiothoracic ratio was 67% and the ejection fraction 13%. Echocardiography showed a dilated left ventricle and thin myocardium. After thorough study, we made a diagnosis of dilated cardiomyopathy. Because conventional therapy was unsuccessful, we conducted partial left ventriculectomy with Alfieri repair of the mitral valve. The postoperative cardiothoracic ratio was 57% at 1 year of follow-up and the ejection fraction 40%. The New York Heart Association functional class improved from IV to I. In conclusion, the role of partial left ventriculectomy is both as a bridge to transplantation and as a definitive repair in dilated cardiomyopathy during childhood. PMID- 11030132 TI - Combined partial sternectomy for thymic carcinoma. AB - For thymic carcinoma, a 67-year-old Japanese female underwent combined chemotherapy, to which a partial response was registered. Afterwards, total thymectomy with combined en bloc resection of the biopsy site, the partial sternum and the partial mediastinal components were completed with a clear margin. Skeletal reconstruction was performed using polypropylene mesh. Paradoxical movement of the chest wall was tolerable. Functional and cosmetic appearance of the chest wall following the reconstruction was satisfactory. Thirty-nine months had passed since the onset of the chemotherapy, when local recurrence was detected in the approximated major pectoral muscle. The patient underwent salvage radiation therapy that resulted in a complete response, and is currently alive and disease free at 46 months after the onset of the combined chemotherapy. PMID- 11030134 TI - Regression of left main coronary ostium stenosis after surgical revascularization and steroid therapy. AB - We describe a patient in whom stenosis of the left main coronary ostium completely regressed after steroid treatment following surgical revascularization. A 46-year-old woman with unstable angina underwent double coronary artery bypass grafting. Although she did not fulfill diagnostic criteria for Takayasu's disease, we began postoperative steroid therapy on postoperative day 14 based on clinical manifestations and histological findings. Coronary angiography 33 days after surgery showed the ostial stenosis of the left main coronary artery had disappeared. Steroid therapy for suspected Takayasu's disease should be considered even after surgical revascularization. PMID- 11030135 TI - Inverted left atrial appendage. AB - An unexpected left atrial mass was found during postoperative echocardiography in a 17-years old man following aortic valve replacement, suggesting thrombus, vegetation and neoplasm. Reoperation showed that the mass to be an inverted left atrial appendage. Left ventricular venting may cause such inversion. To prevent this complication, we suggest the use of careful monitoring and intraoperative transesophageal echocardiography. PMID- 11030136 TI - Adult aortic arch atresia. AB - Atresia in the aortic arch is a rare and severe congenital cardiovascular anomaly. Without surgical therapy, only a few patients can survive to adulthood. A 29-year-old woman with atresia of the aortic arch (Celoria-Patton Type A) without any intracardiac shunt underwent primary surgical correction involving reconstruction of the aortic arch with prosthetic interposition between the transverse aortic arch and the descending aorta and division of the persistent ductus arteriosus. Her postoperative course was uneventful, and she is now doing well at three years to date after surgical treatment. PMID- 11030137 TI - Less invasive therapy using endovascular stent graft repair and video-assisted thoracoscopic surgery for ruptured acute aortic dissection. AB - We report a 75-year-old man with a ruptured acute thoracic aortic dissecting hematoma treated using endovascular stent grafting and video-assisted thoracoscopic surgery. This less invasive therapy is a good therapeutic option even in ruptured acute aortic dissections, particularly given the difficulty of surgery. PMID- 11030138 TI - Primary osteosarcoma of heart with severe congestive heart failure. AB - We present a case report on a 54-year-old woman with extraskeletal osteosarcoma of the left atrium featuring severe congestive heart failure. We resected the tumor, which occupied the left atrium and had widely infiltrated the atrial wall, but the patients died of the tumor 9 months after surgery. This is to our knowledge the 32nd case of cardiac osteosarcoma ever reported. PMID- 11030139 TI - Pericardioperitoneal drainage. PMID- 11030140 TI - Thumb digital neuropathy caused by splinting. AB - Healthcare professionals are currently faced with a great variety of splints and splinting materials. Choices range from prefabricated products to custom splints made on-site from plaster, orthoplast, or fiberglass. In addition to providing immobilization to maintain a particular posture, a splint must protect important soft tissues. Patients with hand or wrist injuries often receive a prefabricated metal cock-up wrist splint in emergency departments. Complications from splints are not uncommon but are infrequently reported. We report a case in which a metal wrist cock-up splint caused compression of the thumb ulnar digital nerve. Preventive measures for such complication are included. PMID- 11030141 TI - An overview of women seeking substance abuse treatment in Oklahoma. AB - Current studies of neurocognitive function in alcoholics are generally restricted to a relatively small subsample of persons seeking treatment. Subjects are typically excluded from study if reporting signs or symptoms of possibly confounding disorders such as comorbid psychiatric disorders, medical, or neurologic disorders. Thus, the question arises as to whether those individuals who actually participate in these reported studies are representative of the larger population of unselected persons. Studies of women are particularly subject to this concern due to higher rates of comorbid medical and psychiatric disorders in women. The current study was directed to contrasting treatment seeking women who either were or were not selected for a laboratory study of neurocognitive function. Specifically, we compared subgroups of women identified on the basis of the presence of an alcohol use disorder and/or being selected for study. A total of 638 women were available for comparison. Contrary to anticipated concerns, analysis of basic demographic, alcohol and drug use, and affective/cognitive measures revealed considerable similarities among the groups. Consistent with other literature, the large majority of these women reported a family history of alcohol use disorders. They also reported a preference for stimulants as opposed to other illicit substances as evidenced in self-report of their most frequently used drug in the six months prior to treatment (excluding alcohol). These findings suggest that current selection procedures, although necessarily biasing samples in response to specific questions, do not result in samples that are significantly different from the majority of treatment-seeking women, at least on the basis of demographic and affective variables. Furthermore, as described herein, these findings have specific indications for primary care assessment and treatment referral. PMID- 11030142 TI - The early years: Jolly West and the University of Oklahoma Department of Psychiatry. PMID- 11030143 TI - APICON-1999. Association of Physicians of India 54th joint annual conference. Calcutta, January 17-20, 1999. Abstracts. PMID- 11030144 TI - p55CDC/hCDC20 is associated with BUBR1 and may be a downstream target of the spindle checkpoint kinase. AB - Eukaryotic cells have evolved a mechanism that delays the progression of mitosis until condensed chromosomes are properly positioned on the mitotic spindle. We have been studying genes that regulated the spindle checkpoint in human cells. Enforced expression of human BUBR1, but not a BUBR1 mutant allele, enhances accumulation of mitotic cells. Yeast two-hybrid system and GST-pulldown analyses show that p55CDC/hCdc20, a protein known to link spindle checkpoint components such as MAD2 to anaphase promoting complex (APC), interacts with BUBR1. In addition, p55CDC is capable of pulling down BUBR1 in sf-9 cells infected with both p55CDC and His6-BUBR1 recombinant baculoviruses but not in the cells infected with p55CDC baculoviruses or with the baculoviral vector alone. Moreover, immunoprecipitation followed by Western blot analyses confirmed that native p55CDC is associated with BUBR1 in HeLa cells. Spindle checkpoint activation by nocodazole treatment enhances the association between p55CDC and His6-BUBR1. In nocodazole-arrested mitotic cells, both CDC16 and hyperphosphorylated CDC27, two APC components, preferentially associate with His6 BUBR1 resins, but not the control resins. Furthermore, BUBR1 phosphorylates p55CDC in vitro, and the phosphorylation of p55CDC by BUBR1 appears to be correlated with spindle checkpoint activation. Together, our studies strongly suggest that BUBR1 may target APC via p55CDC. PMID- 11030145 TI - Caspase-8/FLICE functions as an executioner caspase in anticancer drug-induced apoptosis. AB - Caspase-8 plays an essential role in apoptosis triggered by death receptors. Through the cleavage of Bid, a proapoptotic Bcl-2 member, it further activates the mitochondrial cytochrome c/Apaf-1 pathway. Because caspase-8 can be processed also by anticancer drugs independently of death receptors, we investigated its exact role and order in the caspase cascade. We show that in Jurkat cells either deficient for caspase-8 or overexpressing its inhibitor c-FLIP apoptosis mediated by CD95, but not by anticancer drugs was inhibited. In the absence of active caspase-8, anticancer drugs still induced the processing of caspase-9, -3 and Bid, indicating that Bid cleavage does not require caspase-8. Overexpression of Bcl-x(L) prevented the processing of caspase-8 as well as caspase-9, -6 and Bid in response to drugs, but was less effective in CD95-induced apoptosis. Similar responses were observed by overexpression of a dominant-negative caspase-9 mutant. To further determine the order of caspase-8 activation, we employed MCF7 cells lacking caspase-3. In contrast to caspase-9 that was cleaved in these cells, anticancer drugs induced caspase-8 activation only in caspase-3 transfected MCF7 cells. Thus, our data indicate that, unlike its proximal role in receptor signaling, in the mitochondrial pathway caspase-8 rather functions as an amplifying executioner caspase. PMID- 11030147 TI - Structure and function of the C-terminal hypervariable region of K-Ras4B in plasma membrane targetting and transformation. AB - The C-terminal hypervariable domain of K-Ras4B targets the protein to the plasma membrane by a combination of positive charge and a hydrophobic signal (farnesyl group). We analysed the contribution of several structural features of the domain: net charge, charge distribution, amino acid sequence and lipid specificity to membrane targetting and function by using artificial 'hypervariable' domains fused to either EGFP or V12KRas4B. We found that charge and a lipid residue are sufficient for plasma membrane localization and function of the constitutively active V12K-Ras4B. However, the amount of net charge, charge distribution and the length of the anchoring domain are important. Increasing the net charge and concentrating it close to the C-terminus increases not only the percentage of membrane bound protein, but also shifts the distribution from internal membranes, including the nuclear envelope, to the plasma membrane. While plasma membrane binding is necessary for V12K-Ras4B activity (MAPK activation and focus formation), we found that there are additional restrictions. In particular, mutants with very highly charged domains that bind almost exclusively to the plasma membrane show less transforming potential than expected. In addition, a construct with a short 'hypervariable' domain (7 amino acids) also has decreased transformation activity. These results suggest that specific interactions between K-Ras4B and the plasma membrane are required. PMID- 11030146 TI - Phosphorylation and nuclear exclusion of the forkhead transcription factor FKHR after epidermal growth factor treatment in human breast cancer cells. AB - Akt, when activated by IGF/insulin, can phosphorylate forkhead transcription factors. We undertook this study to determine whether epidermal growth factor (EGF) treatment could produce a signaling cascade resulting in phosphorylation of the forkhead transcription factor FKHR in a breast cancer cell line, MDA-MB-231. After establishing ErbB1, cbl, PI3 kinase and Akt were activated in EGF treated MDA-MB-231, we determined by immunoblot with FKHR antiserum that the electrophoretic mobility of FKHR was retarded after EGF treatment. This mobility retardation was reversible by treatment with alkaline phosphatase, and immunoblot with phospho-Ser256 FKHR antibody further confirmed phosphorylation on an Akt consensus site after EGF treatment. EGF stimulated FKHR phosphorylation was blocked by the PI3 kinase inhibitor LY294002, and the ErbB1 inhibitor AG1478. FKHR immunoblotting after purification of nuclear and cytoplasmic proteins showed that EGF induced a simultaneous increase of FKHR in the cytoplasm and decrease in the nucleus. This finding was confirmed by immunofluorescence staining. Treatment of cells with pharmacological inhibitors of PI3 kinase or ErbB1 blocked this effect. Thus, these results demonstrate the phosphorylation and nuclear exclusion of FKHR after EGF treatment by a PI3 kinase dependent mechanism, and represent the first report of growth factor regulation of endogenous FKHR localization. PMID- 11030148 TI - Activation of the alpha2beta1 integrin prevents c-erbB2-induced scattering and apoptosis of human mammary epithelial cells in collagen. AB - Constitutive overexpression of c-erbB2 in the mammary epithelial cell line MTSV1 7 has been shown to result in epithelial-mesenchymal conversion, anchorage independent growth and loss of organized morphogenesis in collagen. To elucidate the events leading to this drastic change, MTSV1-7 cells and its subclone HB2 (which shows a more strictly epithelial phenotype) were transfected with the hybrid trk-neu receptor consisting of the extracellular domain of the trkA nerve growth factor (NGF) receptor and the transmembrane and cytoplasmic domains of c erbB2 (neu). In cells expressing this construct, c-erbB2 homodimerization can be mimicked by addition of NGF. In trk-neu transfectants of HB2 cells, modest expression led to increased cell proliferation upon NGF treatment. When clones with higher expression levels were grown in collagen, NGF instead induced cell scattering, diminished viability and dramatically increased apoptosis. Interestingly, both the dissociation of colonies and loss of cell viability could be completely reversed by treatment of the cells with antibodies that activate the adhesive capacity of the alpha2beta1 integrin. Long-term NGF treatment of high-expressing transfectants generated fibroblastic clones displaying a reduced expression of integrin alpha2 and E-cadherin, and extensive apoptosis in collagen. These results, which indicate that strong c-erbB2 signalling may lead to downregulation and/or inactivation of the alpha2beta1 integrin, promoting apoptosis in collagen, provide one possible explanation to the increased apoptosis frequently seen in early tumour development. PMID- 11030149 TI - Resistance to TRAIL-induced apoptosis in primitive neuroectodermal brain tumor cells correlates with a loss of caspase-8 expression. AB - TNF-related apoptosis-inducing ligand (TRAIL) is a potent inducer of apoptosis in adult malignant glioma and various other human solid tumor models but not in normal tissues. To characterize the TRAIL death pathway in childhood primitive neuroectodermal brain tumor (PNET), 8 human PNET cell lines were tested for TRAIL induced apoptosis. TRAIL-sensitivity of the PNET cell lines was correlated with mRNA expression levels of TRAIL, its agonistic (TRAIL-R1, TRAIL-R2) and antagonistic (TRAIL-R3, TRAIL-R4) receptors, cellular FLICE-like inhibitory protein (cFLIP), caspase-3 and caspase-8. Three of 8 PNET cell lines tested were susceptible to TRAIL-induced apoptosis. Sensitivity to TRAIL-induced apoptosis did not correlate with mRNA expression of TRAIL receptors or cFLIP. However, all TRAIL-sensitive PNET cell lines expressed caspase-8 mRNA and protein, while none of the five TRAIL-resistant PNET cell lines expressed caspase-8 protein. Treatment with the methyltransferase inhibitor 5-aza-2'-deoxycytidine restored mRNA expression of caspase-8 and TRAIL-sensitivity in formerly TRAIL-resistant PNET cells, suggesting that gene methylation inhibits caspase-8 transcription in these cells. We conclude, that loss of caspase-8 mRNA is an important mechanism of TRAIL-resistance in PNET cells. Treatment with recombinant soluble TRAIL, possibly in combination with methyltransferase inhibitors, represents a promising therapeutic approach for PNET that deserves further investigation. PMID- 11030150 TI - Oncogenes and tumor angiogenesis: the HPV-16 E6 oncoprotein activates the vascular endothelial growth factor (VEGF) gene promoter in a p53 independent manner. AB - Like other types of pre-malignant lesions and carcinoma, angiogenesis is associated with high-grade cervical dysplasia and with invasive squamous carcinoma of the cervix. Vascular endothelial cell growth factor (VEGF) is known to be one of the most important inducers of angiogenesis and is upregulated in carcinoma of the cervix. Human Papilloma Virus 16 (HPV-16) has been etiologically linked to human cervical cancer, and the major oncogenic proteins encoded by the viral genome, E6 and E7, are involved in the immortalization of target cells. Because several oncogenes including mutant ras, EGF receptor, ErbB2/Her2, c-myc and v-src upregulate VEGF expression, we asked whether HVP-16 E6 oncoprotein could act in a similar fashion. We found that HPV-16 E6-positive cells generally express high levels of VEGF message. Furthermore, co-expression of the VEGF promoter-Luc (luciferase) reporter gene with E6 in both human keratinocytes and mouse fibroblast showed that E6 oncoprotein upregulates VEGF promoter activity, and does so in a p53 independent manner. An E6 responsive region which comprises four Sp-1 sites, between -194 and -50 bp of the VEGF promoter, is also necessary for constitutive VEGF transcription. Taken together, our results suggest the possibility that the HPV oncoprotein E6 may contribute to tumor angiogenesis by direct stimulation of the VEGF gene. PMID- 11030151 TI - Hypoxia-induced VEGF enhances tumor survivability via suppression of serum deprivation-induced apoptosis. AB - Low oxygen and nutrient depletion play critical roles in tumorigenesis, but little is known about how they interact to produce tumor survival and tumor malignancy. In the present study, we investigated the mechanism underlying hypoxia-modulated apoptosis of serum-deprived HepG2 cells. Our results showed that hypoxia blocked the apoptosis, which was accompanied with decreased Bax/Bcl 2 ratio, inhibited cytochrome c release, and reduced caspase-3 activity. More importantly, increased expressions of VEGF and its receptor-2 (KDR) under hypoxic/serum-deprived condition suggest that VEGF may act as a survival factor in a self-promoting manner. Data were further supported by results that recombinant human VEGF (rhVEGF) suppressed the serum deprivation-induced apoptosis, and anti-VEGF neutralizing antibody block anti-apoptotic activity of hypoxia. In addition, inhibitors of receptor tyrosine kinase blocked antiapoptosis of hypoxia. Our study further showed that rhVEGF or hypoxia induced ERK phosphorylation in serum-deprived cells, and that a specific inhibitor of MAPK/ERK, PD98059 eliminated the anti-apoptotic activity of rhVEGF or hypoxia by increasing Bax/Bcl-2 ratio and caspase-3 activity. Our data led us to conclude that induction of ERK phosphorylation and decrease of Bax/Bcl-2 ratio by rhVEGF implies that hypoxia-induced VEGF prevents apoptosis of serum-deprived cells by activating the MAPK/ERK pathway. Taken together, we propose that hypoxia enhances survival of nutrient-depleted tumor cells by reducing susceptibility to apoptosis, which consequently leads to tumor malignancy. PMID- 11030152 TI - Protein expression of the RB-related gene family and SV40 large T antigen in mesothelioma and lung cancer. AB - Mutational inactivation of the RB-related gene RBL2/p130 has been reported as a common and important prognostic factor in human lung cancer. To examine the role of the RB-related gene family in lung cancer we analysed the protein expression of the RB gene in cell lines obtained from 83 patients with small cell lung cancer (SCLC) and 114 patients with non-SCLC that included 21 novel lung tumor samples. While we detected five new SCLC with mutant RB expression (RB inactivation in 75/83; 90.4%), we did not detect any RB mutations in the new non SCLC cell lines (RB inactivation in 13/114 non-SCLC and mesothelioma; 11.4%). In addition, we detected expression of a full-length RBL1/p107 and RBL2/p130 species in every sample tested (RBL1 or RBL2 inactivation in 0/69) and confirmed that both RB-related gene products retain functional binding activity to the E1A viral oncoprotein. Since expression of SV40 Large T antigen (Tag) has been reported in a subset of human lung tumors where it may inactivate RBL1 and RBL2, we also examined mesothelioma and non-mesothelioma lung tumors for Tag expression. Although we detected a faint 85 kDa protein species using specific anti-Tag antibodies, this signal migrated slightly faster than Tag extracted from Cos7 cells and did not exhibit binding activity to the RB or RBL1 proteins. Finally, we subjected 11 lung cancer cell lines to nucleotide sequencing and did not detect mutations within the C-terminal RBL2 exons 19-22 as recently reported. While the RB/p16 tumor suppressor pathway is targeted for mutations in 100% of lung cancers, mutational inactivation of the related RBL1 and RBL2 genes is a rare event. PMID- 11030153 TI - Complex regulation of CDKs and G1 arrest during the granulocytic differentiation of human myeloblastic leukemia ML-1 cells. AB - We previously reported that all-trans retinoic acid (ATRA) and granulocyte macrophage colony-stimulating factor (GM-CSF) synergistically induced granulocytic differentiation in human myeloblastic leukemia ML-1 cells. The combination of these agents also suppressed DNA-synthesis. In the present study, we investigated the suppression of cyclin dependent kinase (CDK) activities resulting in G1 arrest in differentiated ML-1 cells. We show that treatment of ML 1 cells with ATRA plus GMCSF results in G1 arrest and suppression of CDK activities. Protein levels of the G1 CDKs were essentially unchanged during this time. However, we observed an increase in CDK2-bound p27 and CDK4-bound p18, and a decrease in CDK6-bound cyclin D3. These results suggest that complex regulation of CDKs play a key role in G1 arrest of ML-1 after treatment with ATRA and GM CSF. We also showed that an increase in CDK2-bound p27 and CDK4-bound p18 are caused by treatment with ATRA and a decrease in CDK6-bound cyclin D3 is induced synergistically by treatment with both reagents. Furthermore, we propose that the changes in binding of p18 and cyclin D3 to CDKs are due to changes at the protein expression level and that the increase in p27 binding to CDK2 is due to a novel mechanism. PMID- 11030154 TI - MGSA/GRO-mediated melanocyte transformation involves induction of Ras expression. AB - The MGSA/GRO protein is endogenously expressed in almost 70% of the melanoma cell lines and tumors, but not in normal melanocytes. We have previously demonstrated that over-expression of human MGSA/GROalpha, beta or gamma in immortalized murine melanocytes (melan-a cells) enables these cells to form tumors in SCID and nude mice. To examine the possibility that the MGSA/GRO effect on melanocyte transformation requires expression of other genes, differential display was performed. One of the mRNA's identified in the screen as overexpressed in MGSA/GRO transformed melan-a clones was the newly described M-Ras or R-Ras3 gene, a member of the Ras gene superfamily. Over-expression of MGSA/GRO upregulates M Ras expression at both the mRNA and protein levels, and this induction requires an intact glutamine-leucine-arginine (ELR)-motif in the MGSA/GRO protein. Western blot examination of Ras expression revealed that K- and N-Ras proteins are also elevated in MGSA/GRO-expressing melan-a clones, leading to an overall increase in the amount of activated Ras. MGSA/GRO-expressing melan-a clones exhibited enhanced AP-1 activity. The effects of MGSA/GRO on AP-1 activation could be mimicked by over-expression of wild-type M-Ras or a constitutively activated M Ras mutant in control melan-a cells as monitored by an AP-1-luciferase reporter, while expression of a dominant negative M-Ras blocked AP-1-luciferase activity in MGSA/GRO-transformed melan-a clones. In the in vitro transformation assay, over expression of M-Ras mimicked the effects of MGSA/GRO by inducing cellular transformation in control melan-a cells, while over-expression of dominant negative M-Ras in MGSA/GROalpha-expressing melan-a-6 cells blocked transformation. These data suggest that MGSA/GRO-mediated transformation requires Ras activation in melanocytes. PMID- 11030155 TI - Repression of transforming growth factor-beta receptor type I promoter expression by Sp1 deficiency. AB - In this report, we describe the mechanism of TGF-beta receptor type I (RI) repression in the GEO human colon carcinoma cells. Treatment of GEO cells with the DNA methyltransferase inhibitor, 5 azacytidine induced RI expression and restored TGF-beta response. A stably transfected RI promoter-reporter construct (RI-Luc) expressed higher activity in the 5 aza C treated GEO cells, suggesting the activation of a transactivator for RI transcription. Gel shift analysis indicated enhanced binding of proteins from the 5 aza C treated nuclear extracts to radiolabeled Sp1 oligonucleotides specifically contained in the RI promoter. Protein stability studies after cyclohexamide treatment suggested an increase in the Sp1 protein stability from the 5 aza C treated GEO cells. Further, transfection of Sp1 cDNA into untreated GEO control cells increased RI promoter activity and thus induced RI expression. 5 aza C mediated Sp1 expression in Sp1 deficient GEO colon and MCF-7 breast cancer cells also enhanced the activity of several other Sp1 dependent promoters such as TGF-beta receptor type II (RII), Cyclin A and p21/waf1/cip1. These results indicate that restoration of Sp1 in several different types of Sp1 deficient cells leads to enhanced activation of a wide range of Sp1 dependent promoters. PMID- 11030156 TI - Evaluation of organ failure: we are making progress. PMID- 11030157 TI - Assessment of intestinal tissue oxygenation: the canary sings--but was does the twitter tell us? PMID- 11030158 TI - Cerebrovenous oxygen saturation monitoring: practical considerations and clinical relevance. AB - Measurement of the saturation of brain effluent blood gives a global estimate of cerebral oxygenation. It may provide clinicians with information to assist in reducing secondary insults to the brain with potential benefits to a range of patients with actual or potential acute brain injury such as trauma and cardiac bypass procedures. The technology to continuously measure this variable is simple to use but requires attention to detail; it is limited in its ability to detect discrete regions of ischaemia or hyperaemia unless these are of sufficient magnitude to influence the saturation of brain effluent blood. There are few complications that result from this invasive technique and they are usually of a minor nature. The technique also enables research opportunities from the ability to sample blood as it leaves the cranium. Poor outcomes are seen in patients with traumatic brain injury who exhibit either reduced or increased cerebrovenous oxygen saturation and it remains to be seen if detection and correction of these anomalies will produce patient benefits. PMID- 11030159 TI - Evaluation of the SOFA score: a single-center experience of a medical intensive care unit in 303 consecutive patients with predominantly cardiovascular disorders. Sequential Organ Failure Assessment. AB - OBJECTIVE: To evaluate the use of the Sequential Organ Failure Assessment (SOFA) score, the total maximum SOFA (TMS) score, and a derived variable, the deltaSOFA (TMS score minus total SOFA score on day 1) in medical, cardiovascular patients as a means for describing the incidence and severity of organ dysfunction and the prognostic value regarding outcome. DESIGN: Prospective, clinical study. SETTING: Medical intensive care unit in a university hospital. PATIENTS: A total of 303 consecutive patients were included (216 men, 87 women; mean age 62 +/- 12.6 years; SAPS II 26.2 +/- 12.7). They were evaluated 24 h after admission and thereafter every 24 h until ICU discharge or death between November 1997 and March 1998. Readmissions and patients with an ICU stay shorter than 12 h were excluded. MAIN OUTCOME MEASURE: Survival status at hospital discharge, incidence of organ dysfunction/failure. INTERVENTIONS: Collection of clinical and demographic data and raw data for the computation of the SOFA score every 24 h until ICU discharge. MEASUREMENTS AND MAIN RESULTS: Length of ICU stay was 3.7 +/ 4.7 days. ICU mortality was 8.3% and hospital mortality 14.5%. Nonsurvivors had a higher total SOFA score on day 1 (5.9 +/- 3.7 vs. 1.9 +/- 2.3, p < 0.001) and thereafter until day 8. High SOFA scores for any organ system and increasing number of organ failures (SOFA score > or = 3) were associated with increased mortality. Cardiovascular and neurological systems (day 1) were related to outcome and cardiovascular and respiratory systems, and admission from another ICU to length of ICU stay. TMS score was higher in nonsurvivors (1.76 +/- 2.55 vs. 0.58 +/- 1.39, p < 0.01), and deltaSOFA/total SOFA on day 1 was independently related to outcome. The area under the receiver-operating characteristic curve was 0.86 for TMS, 0.82 for SOFA on day 1, and 0.77 for SAPS II. CONCLUSIONS: The SOFA, TMS, and deltaSOFA scores provide the clinician with important information on degree and progression of organ dysfunction in medical, cardiovascular patients. On day 1 both SOFA score and TMS score had a better prognostic value than SAPS II score. The model is closely related to outcome and identifies patients who are at increased risk for prolonged ICU stay. PMID- 11030160 TI - Regional distribution of gas and tissue in acute respiratory distress syndrome. II. Physiological correlations and definition of an ARDS Severity Score. CT Scan ARDS Study Group. AB - OBJECTIVES: (a) To assess whether differences in lung morphology observed in patients with adult respiratory distress syndrome (ARDS) are associated with differences in cardiorespiratory parameters, lung mechanics, and outcome. (b) To propose a new ARDS Severity Score to identify patients with a high mortality risk. DESIGN: Prospective study over a 53-month period. SETTING: Fourteen-bed surgical intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: Seventy-one consecutive patients with early ARDS. MEASUREMENTS AND RESULTS: Cardiorespiratory parameters were measured using a Swan-Ganz catheter, the pressure-volume (PV) curve was measured using the gross syringe method, and fast spiral computed tomography (CT) was performed. Patients with diffuse attenuations (n = 16) differed from patients with lobar attenuations (n = 26) regarding: (a) mortality rate (75% vs. 42%, p = 0.05), (b) incidence of primary ARDS (82% vs. 50%, p = 0.03), (c) respiratory compliance (47 +/- 12 vs. 64 +/- 16 ml per cmH2O( 1) p = 0.04), and (d) lower inflexion point (8.4 +/- 2.0 vs. 4.6 +/- 2.0 cmH2O, p = 0.001). A third group of patients with patchy attenuations (n = 29) had a mortality rate of 41 %, a respiratory compliance of 56 +/- 18 ml per cmH2O(-1) and a lower inflexion point of 6.3 +/- 2.7 cmH2O. The bedside chest radiograph accurately assessed lung morphology in only 42% of the patients. In contrast to the scores based on the bedside chest radiograph, a new ARDS Severity Score based on CT lung morphology and cardiorespiratory parameters identified a subgroup of patients with a high mortality rate (> or = 60%). CONCLUSIONS: In patients with ARDS, differences in lung morphology are associated with differences in outcome and lung mechanics. A new ARDS Severity Score based on CT lung morphology and cardiorespiratory parameters accurately identified patients with the most severe forms of ARDS and a mortality rate above 60%. PMID- 11030161 TI - Linear and nonlinear analysis of pressure and flow during mechanical ventilation. AB - OBJECTIVE: Linear modeling as a method of exploring respiratory mechanics during mechanical ventilation, was compared to nonlinear modeling for flow dependence of resistance in three distinct groups of patients, those with: (a) normal respiratory function (NRF), (b) chronic obstructive pulmonary disease (COPD), or (c) adult respiratory distress syndrome (ARDS). DESIGN AND PATIENTS: Airways opening pressure (Pao), flow (V'), and volume (V) signals were recorded in 32 ICU mechanically ventilated patients, under sedation and muscle relaxation (10 NRF, 11 COPD, 11 ARDS). All patients were ventilated with controlled mandatory ventilation mode at three levels of end-expiratory pressure (PEEPe): 0, 5, and 10 hPa. Data were analyzed according to: (a) Pao = PE + Ers V + Rrs V' and (b) Pao = PE + Ers V + k1V' + k2?V'?V'; where Ers and Rrs represent the intubated respiratory system (RS) elastance and resistance, k1 and k2 the linear and the nonlinear RS resistive coefficients, and PE the end-expiratory pressure. The model's goodness of fit to the data was evaluated by the root mean square difference of predicted minus measured Pao values. RESULTS: NRF data fit both models well at all PEEPe levels. ARDS and particularly COPD data fit the nonlinear model better. Values of k2 were often negative in COPD and ARDS groups, and they increased in parallel with PEEPe. A gradual increase in PEEPe resulted in better fit of ARDS and COPD data to both models. CONCLUSIONS: The model of V' dependence of resistance is more suitable for the ARDS and particularly the COPD groups. PEEP tends to diminish the V' dependence of respiratory resistance during the respiratory cycle, particularly in the COPD group, probably through an indirect effect of the increased lung volume. PMID- 11030162 TI - Anterior magnetic phrenic nerve stimulation: laboratory and clinical evaluation. AB - OBJECTIVE: Anterior magnetic stimulation (aMS) of the phrenic nerves is a new method for the assessment of diaphragm contractility that might have particular applications for the clinical assessment of critically ill patients who are commonly supine. DESIGN: We compared aMS with existing techniques for measurement of diaphragm weakness and fatigue in 10 normal subjects, 27 ambulant patients with suspected diaphragm weakness and 10 critically ill patients. SETTING: Laboratory and intensive care unit of two university hospitals. RESULTS: Although aMS was not demonstrably supramaximal in normal subjects, the mean value of twitch transdiaphragmatic pressure (Tw Pdi) obtained at 100% of stimulator output, 23.7 cmH2O, did not differ significantly from that obtained with bilateral supramaximal electrical stimulation (ES), 24.9 cmH2O, or bilateral anterior magnetic phrenic nerve stimulation (BAMPS), 27.3 cmH2O. A fatiguing protocol produced a 20 % fall in aMS-Tw Pdi and a 19% fall in BAMPS-Tw Pdi; the fall in aMS-Tw Pdi correlated with the fall in BAMPS-Tw Pdi (r2 = 0.84, p = 0.03) indicating that aMS can detect diaphragm fatigue. In ambulant patients aMS agreed closely with existing measures of diaphragm strength. The maximal sniff Pdi correlated with both the aMS-Tw Pdi (r2 = 0.60, p < 0.0001) and the BAMPS-Tw Pdi (r2 = 0.65, p < 0.0001) and the aMS-Tw Pdi was a mean (SD) 2.2 (4.3) cmH2O less than BAMPS-Tw Pdi. In addition, aMS correctly identified diaphragm dysfunction in patients studied on the ICU. CONCLUSIONS: We conclude that aMS is of clinical value for the investigation of suspected diaphragm weakness. PMID- 11030163 TI - Immediate IL-10 expression following major orthopaedic trauma: relationship to anti-inflammatory response and subsequent development of sepsis. AB - OBJECTIVE: To assess the relationship between IL-10 release and anti-inflammatory response following blunt trauma. DESIGN: Prospective longitudinal clinical study. SETTING: Departments of trauma and anaesthetics in a university teaching hospital. PATIENTS: Forty-eight adult patients with a mean injury severity score of 14.5 (range 9-57) were prospectively studied following blunt trauma. MEASUREMENTS AND RESULTS: Venous blood samples were collected on arrival and at 16 and 24 h, and at 3, 5, and 7 days. Peripheral blood mononuclear cell (HLA-DR) expression on CD14 + monocytes was quantified by flow cytometry and serum IL-10 was assayed by ELISA. Anti-inflammatory response was defined as monocyte HLA-DR expression of less than 30% of that seen in healthy controls. Serum IL-10 levels in trauma patients on arrival was significantly elevated, 70.0 [48.0-92.1, 95% confidence interval, (CI)] compared to the control group, 3 (0-5) (P < 0.0001), and monocyte HLA-DR expression was significantly lower, 14.2 (12.1-16.3, 95% CI), in patients versus 25.2 (22.4-28.1) in controls (P < 0.001). Patients with low HLA-DR expression (n = 14) had significantly higher serum IL-10 levels than those whose HLA-DR expression remained above 30% of the control value (n = 34), (P < 0.038). In patients who developed sepsis (n = 11), serum IL-10 levels were greater on admission, [143.7 (80.2-207.2) pg/ml(-1)], and remained elevated during the study period compared with non-complicated patients, [50.16 (33.5 66.8) pg/ml(-1)]. Immediate IL-10 (2 h following trauma) was negatively correlated with simultaneous HLA-DR expression, (r = -0.49, P = 0.0005). CONCLUSION: These findings support the view that IL-10 release regulates monocyte HLA-DR expression and may be related to an anti-inflammatory response and development of sepsis following trauma. PMID- 11030164 TI - Serum and ascitic procalcitonin levels in cirrhotic patients with spontaneous bacterial peritonitis: diagnostic value and relationship to pro-inflammatory cytokines. AB - OBJECTIVE: To assess the potential role of procalcitonin and pro-inflammatory cytokines, TNF-alpha, and IL-6, in the diagnosis of spontaneous bacterial peritonitis (SBP). DESIGN: Prospective study. SETTING: The emergency unit of a teaching hospital. PATIENTS: We included 21 patients with SBP and 40 patients with sterile ascitic fluid. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: For the diagnosis of SBP, the best markers were serum levels of procalcitonin with a cut-off value of 0.75 ng/ml, a sensitivity of 95%, a specificity of 98%, and ascitic fluid levels of IL-6 with a cut-off value of 5,000 ng/ml, a sensitivity of 100%, and a specificity of 88%. C-reactive protein and serum polymorphonuclear count have low sensitivity/specificity at 62/92% and 57/90%, respectively. From 21 patients with SBP, ascitic fluid to serum ratio of TNF alpha and IL-6 was greater than to 2 in all cases with a mean at 6.2 +/- 6.5 and 34 +/- 31, respectively. By contrast, this ratio for procalcitonin was less than 1 in all cases with a mean at 0.31 +/- 0.25. We found no correlation between procalcitonin levels and cytokine levels in either ascitic fluid or serum. CONCLUSIONS: Serum procalcitonin level may become a useful marker for the diagnosis of SBP in cirrhotic patients. The low ratio of ascitic fluid to serum procalcitonin supports the hypothesis that procalcitonin is not produced intraperitoneally. PMID- 11030165 TI - Pharmacokinetics of milrinone in patients with congestive heart failure during continuous venovenous hemofiltration. AB - OBJECTIVE: To evaluate the pharmocokinetics of intravenous milrinone in patients with severe congestive heart failure during continuous venovenous hemofiltration (CVVH). DESIGN: Prospective study of patients with congestive heart failure admitted to the intensive care unit (ICU). SETTING: ICU between September 1997 and August 1999. PATIENTS AND METHODS: Six patients with severe congestive heart failure during CVVH: all patients received a continuous infusion of milrinone of 0.25 microg x kg(-1) min(-1). The hemodynamics and plasma concentration of milrinone were measured before and after the infusion. Pharmacokinetics were analyzed with one-compartment model featuring constant rate infusion. RESULTS: The steady-state concentration (Css) was 845 +/- 135 (mean +/- SD) ng/ml, and the half-life time (t1/2) was 20.1 +/- 3.3 h. Cardiac index and stroke volume index after the infusion of milrinone increased significantly compared with pre infusion levels. Other hemodynamic parameters did not change significantly. All patients died within 1 month after the injection of milrinone because of severe forms of arrhythmia, such as ventricular tachycardia and ventricular fibrillation. CONCLUSIONS: We found that the mean Css and the mean t1/2 of milrinone in subjects during CVVH were much higher and longer than those previously reported for subjects with normal renal function. It is therefore essential to adjust the dose or modify the dosing interval of milrinone during renal replacement therapy for patients with severe congestive heart failure. However, further studies are needed to determine the details of pharmacokinetics of milrinone and therapeutic procedures for patients with severe heart failure during CVVH. PMID- 11030166 TI - Cost analysis of plasma-exchange therapy for the treatment of Guillain-Barre syndrome. French Cooperative Group on Plasma Exchange in Guillain-Barre Syndrome. AB - OBJECTIVE: To undertake a cost analysis of therapeutic strategies with plasma exchange (PE) for the treatment of patients with Guillain-Barre syndrome. DESIGN: A randomized clinical trial including 556 patients with Guillain-Barre syndrome. We demonstrated that in the group with mild disease (walking possible) two PEs were more effective than none in shortening the time to beginning motor recovery. In the groups with moderate disease (walking impossible) and or severe disease (mechanically ventilated patients) four sessions were more effective than two and no more effective than six in shortening the time to recovery of walking with assistance and for the recovery rate of full muscle strength within 1 year. Data on outcomes and costs was collected. Complete cost data were available on 546 from the 556 patients of the trial. Costs were estimated from the viewpoint of the healthcare system and computed over a 1-year period. Because the analysis of medical outcomes did not show any difference regarding mortality but only on intermediate short-term and long-term outcomes, we carried out a cost minimization analysis. RESULTS: In two groups a dominant strategy appeared, with greater efficacy and lower costs in the two-PE arm for the mild group: 21,353 euros vs. 38,753 euros and in the four-PE arm in the moderate group: 59,480 euros vs. 80,737 euros. In the severe group four PEs were as efficient and somewhat less expensive than six: 57,621 vs. 61,056 euros. CONCLUSION: The treatment of Guillain-Barre syndrome by PE at the onset of disease appears to have medical justification. The least expensive strategies are either more or equally efficient as more expensive strategies. PMID- 11030167 TI - Early translaryngeal tracheostomy in patients with severe brain damage. AB - OBJECTIVES: To describe the effects of early translaryngeal tracheostomy on intracranial pressure (ICP), cerebral perfusion pressure (CPP), and jugular bulb saturation (SjO2); to identify the main mechanisms affecting ICP during tracheostomy; and to evaluate the long-term effects of tracheostomy on tracheal anatomy and function. DESIGN: Prospective, observational, clinical study. SETTING: Neurosurgical intensive care unit in a teaching hospital. PATIENTS: 20 patients admitted to the ICU because of head injury, subarachnoid hemorrhage, or brain tumor with a Glasgow Coma Scale less than 8. INTERVENTIONS: Patients underwent translaryngeal tracheostomy under strict neuromonitoring. MEASUREMENTS AND RESULTS: ICP rose significantly (p < 0.05) at the critical time of cannula placement while all other parameters remained stable. At this time five patients suffered intracranial hypertension (ICP > 20 mmHg). In one of them CPP dropped below 60 mmHg. Arterial CO2 tension (PaCO2) did not rise significantly. No other major complications were recorded during the procedures. Three months after tracheostomy normal findings were detected by tracheoscopy in all cases (11 patients could be examined). CONCLUSIONS: Translaryngeal tracheostomy, performed in selected patients when the risk of intracranial hypertension was reduced to the minimum, was well tolerated in the majority of cases and did not induce persistent intracranial disorders. However, ICP is affected by tracheostomy, and careful monitoring and patient selection is necessary. At follow-up no severe anatomical or functional damage was detected. PMID- 11030168 TI - Blood in the gastrointestinal tract delays and blunts the PCO2 response to transient mucosal ischaemia. AB - OBJECTIVE: To determine whether the presence of blood in the gastrointestinal tract impairs the detection of splanchnic mucosal ischaemia by reducing the rate and magnitude of rise in gut luminal PCO2. DESIGN: Prospective observational study. SETTING: University Research Laboratory. SUBJECTS: Four adult male Wistar rats with four controls from a concurrent study. INTERVENTIONS: Four adult male Wistar rats were anaesthetised with sodium pentobarbitone and ventilated with oxygen and isoflurane to an initial PaCO2 of 30-40 torr. Electrochemical fiberoptic gas sensors inserted into silastic tubing placed in the ileal lumen measured luminal PCO2 at 1-s intervals. Distal aortic pressure was monitored continuously. Six 2-min episodes of aortic hypotension were induced in each rat to a mean pressure of 30 mmHg by intermittent elevation of a silk sling placed around the proximal aorta. Before the last three episodes in each rat 0.75-1.0 ml blood was injected into the ileal lumen via a 25-gauge needle. Four control rats from a concurrent experiment were treated in an identical fashion except that the rats were subjected to five ischaemic episodes, and there was no intraluminal injection of blood, although a 20-gauge cannula was placed in the proximal ileal lumen. MEASUREMENTS AND MAIN RESULTS: The presence of blood in the lumen significantly decreased the ischaemic deltaPCO2 response from 33 +/- 8 to 15 +/- 4 torr (P < 0.001) and also significantly increased the peak response time from 188 +/- 12 to 227 +/- 24 s (P < 0.001). The corresponding measurements in the concurrent controls differed only from the values after blood injection in the experimental group. CONCLUSIONS: In this animal model the presence of intraluminal blood significantly delayed the rate and the amplitude of luminal PCO2 increases in response to transient ischaemia. This raises questions about the validity of luminal CO2 measurements as an indicator of splanchnic ischaemia in the presence of gastrointestinal haemorrhage. PMID- 11030169 TI - Theoretical and experimental comparison of constant inspired concentration and pulsed delivery in NO therapy. AB - OBJECTIVE: Inhaled NO therapy of artificially ventilated patients has been established as being based on constant inspired concentration of NO. In this study a new volumetrically controlled pulsed NO delivery mode is compared with the established concentration-based concept. DESIGN: To evaluate the relationship between NO delivery parameters, alveolar NO fraction, and patient uptake, a mathematical lung model was created where NO delivery can be simulated in varying ventilator settings, delivery modes, and lung properties. This model and the efficacy of pulsed delivery in inducing pulmonary capillary vasodilatation were examined experimentally. SETTING: Animal laboratory, Department of Medical Sciences, Clinical Physiology. SUBJECTS: The experimental study was performed with nine pigs of mixed breed weighing 25-35 kg. INTERVENTIONS: The pigs were anaesthetised and artificially ventilated. Pulmonary vasoconstriction was induced by hypoxia. NO was delivered periodically in the various delivery modes. MEASUREMENTS AND RESULTS: In simulation, in all delivery modes the NO uptake was found to be dependent on the ventilator settings and the volume of the dead space. Measured from pulmonary artery pressure, the pulsed delivery was as effective in reducing the induced pulmonary vasoconstriction as the constant inspired concentration delivery. The amount of NO that could reduce the vasoconstriction back to baseline was 105 nmol x min(-1). By delivering in the early part of the inspiration, ambient contamination by the exhaust gas is avoided. The expired NO values obtained in the simulation and the experiments were equal. Based on the simulation, the alveolar NO fraction and the NO uptake depend on the ventilator settings and the dead space in both volumetric- and concentration-based delivery. CONCLUSIONS: With pulsed delivery, a therapeutic effect comparable to constant inspired concentration delivery is achieved, NO gas is used more effectively, and environmental exhausts are reduced. The theoretical model shows that the NO delivery does not predict alveolar NO fraction and the NO uptake. However, it still remains an open question if the online measurement of these parameters would provide useful information, having added value in predicting and controlling the efficacy of the NO treatment. PMID- 11030170 TI - Effects of haemoconcentration and haemodilution on acute hypoxia-induced pulmonary hypertension and changes in vascular compliance of isolated rabbit lungs. AB - OBJECTIVE: Erythrocytes influence the magnitude of hypoxia-induced pulmonary artery pressure increase. It is, however, unknown to what extent haemoconcentration and haemodilution affect this response and whether intrapulmonary blood volume (and thus vessel dimensions) alters the magnitude of pressure increase. Furthermore, it is unclear whether the haemodilution/ haemoconcentration-dependent pressure increase is flow-related, via flow dependent changes in vasomotor tone or rheologic effects, or can also be observed under no-flow conditions. DESIGN: Experimental study in isolated rabbit lungs (n = 12) perfused with autologous blood at constant flow (100 ml/min) and ventilated with 5% carbon dioxide in air. SETTING: Laboratory for experimental studies. INTERVENTIONS: Haemoconcentration (centrifugation) and haemodilution (Krebs Henseleit/albumin) were carried out, resulting in haematocrits between 50% and 0%. During hypoxic ventilation, inspiratory oxygen fraction was reduced from 0.20 to 0.03. MEASUREMENTS AND RESULTS: Under constant flow conditions, haemodilution (from a Hct of 34-36% to 0-1%) decreased hypoxic pulmonary artery pressure response to one-third (from 10.8 +/- 2.3 cmH2O to 3.1 +/- 1.0 cmH2O, P < 0.05), while haemoconcentration did not affect the magnitude of hypoxic response (10.5 +/- 2.0 cmH2O). For all haematocrit values an increase in pulmonary blood volume (by 5 ml) decreased the magnitude of pressure response. Hypoxia-induced changes in static vascular filling pressure (double occlusion pressure) and vascular compliance were used to assess the strength of hypoxic vasoconstriction under static conditions. Neither haemoconcentration nor haemodilution altered hypoxia induced changes in either variable. CONCLUSIONS: The magnitude of the acute hypoxic pressure response is not altered by haemoconcentration, but significantly reduced by haemodilution. In contrast, neither haemoconcentration nor haemodilution influenced hypoxia-induced changes in static vascular filling pressure and compliance. This suggests that the degree of hypoxic pulmonary vasoconstriction is not affected under static conditions and that the red blood cell-dependence of the magnitude of hypoxic pressure response is based on flow related mechanisms. PMID- 11030171 TI - Glucose dilution can detect fluid redistribution following phentolamine infusion in dogs. AB - OBJECTIVE: We have recently reported that the initial distribution volume of glucose (IDVG) reliably measures the central extracellular fluid (ECF) volume in the presence of fluid gain or loss. However, it is not clear if IDVG consistently reflects central-ECF volume when redistribution of fluid occurs in the absence of fluid gain or loss. This study was designed to investigate changes in fluid volumes during phentolamine infusion in dogs. DESIGN: Prospective animal study. SETTING: Institutional animal research laboratory. SUBJECTS: Fourteen anesthetized and ventilated mongrel dogs. INTERVENTIONS: Anesthetized animals were mechanically ventilated and received infusions of normal saline (n = 7) or phentolamine (10 microg kg min) (n = 7). Plasma volume was estimated using the indocyanine green (ICG) dilution method (PV-ICG) and IDVG was calculated using a one-compartment model by simultaneous administration of ICG 0.5 mg/kg and glucose 100 mg/kg before, during, and after infusion of either drug. MEASUREMENTS AND RESULTS: PV-ICG during infusion was not different between groups. However, IDVG significantly decreased (P < 0.05) following phentolamine infusion when compared with normal saline infusion. CONCLUSION: Our results suggest that IDVG rather than PV-ICG consistently measures central extracellular fluid volume, even when redistribution of fluid occurs. PMID- 11030172 TI - The Pstl polymorphism of the endotoxin-inducible heat-shock protein 70-2 gene does not affect messenger RNA level in human whole-blood cultures. AB - OBJECTIVE: To determine whether the human leukocyte antigen linked biallelic heat shock protein 70-2 (HSP70-2) gene polymorphism is associated with variable HSP70 2 messenger RNA expression. DESIGN: Prospective observational study in consecutive healthy blood donors. SETTING: Department of Anesthesiology, laboratory for molecular biology in a university hospital. PARTICIPANTS: 24 healthy blood donors. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: We studied the functional implication of the HSP70-2 (G/A) PstI gene polymorphism in 24 healthy, white blood donors with various HSP70-2 (G/A) genotypes by analyzing the endotoxin-inducible HSP70-2 mRNA expression by means of the reverse transcription polymerase chain reaction. HSP70 expression was expressed semiquantitatively by calculating the ratio of HSP70-2 mRNA and the constitutively expressed glutaraldehyde 3-phosphate dehydrogenase mRNA. No significant differences in HSP70-2 mRNA expression after lipopolysaccharide (from Salmonella minnesota Re 595) stimulation were detected in individuals homozygous for the allele A (0.68, range 0.38-1, n = 10), in individuals homozygous for the allele G (0.79, range 0.42-1.1, n = 8), and in heterozygotes (HSP70-2 G/A; 0.52, range 0.4-0.67, n = 6; p > 0.05). CONCLUSIONS: The PstI polymorphism of the endotoxin-inducible HSP70-2 gene is not associated with variable HSP70-2 mRNA expression ex vivo. This finding is in accordance with the observation that HSP70-2 genotypes do not affect clinical outcome in human systemic inflammation. PMID- 11030173 TI - Coma mimicking brain death following baclofen overdose. AB - Baclofen toxicity can be a cause of profound coma with brainstem dysfunction mimicking brain death, and is mainly a clinical diagnosis. Measuring plasma levels is not always possible and may be misleading. Imaging results are usually normal. Electroencephalography may show a pattern of burst suppression. At present no effective specific therapy is available. However, as demonstrated in our case, the prognosis can be good even in severe cases, provided it is recognized early enough, and appropriate supportive measures are instituted. PMID- 11030174 TI - Inadvertent transpericardial insertion of a central venous line with cardiac tamponade failure of preventive practices. AB - A 56-year-old man who had undergone cardiac surgery suffered from cardiac tamponade after administration of contrast-medium through a central venous catheter. Pericardiotomy showed the catheter transversing the pericardial sac just beneath an unusual high reflection and then reentering the superior vena cava. Preventive practices including chest radiography, confirming free venous blood return and manometry may fail to detect catheter malposition in rare cases. Knowledge of potential pitfalls in using generally recommended safety practices and continuous vigilance are essential for the anesthesiologist and intensivist in avoiding potentially lethal hazards. PMID- 11030175 TI - Lung-protective mechanical ventilation strategies in ARDS. PMID- 11030176 TI - Fever in patients with pacemakers: the necessity of perfoming transesophageal echocardiography early. PMID- 11030177 TI - ARDS as a complication of difficult airway management in a patient with Hunter syndrome. PMID- 11030178 TI - Subarachnoid hemorrhage and acute myocardial infarction. PMID- 11030179 TI - Oesophageal intestinal metaplasia. PMID- 11030180 TI - Manometric and pH-metric features in gastro-oesophageal reflux disease patients with and without Helicobacter pylori infection. AB - BACKGROUND: The role of Helicobacter pylori in the pathogenesis and evolution of gastro-oesophageal reflux disease is still debated. AIM: To investigate the impact of Helicobacter pylori infection on the oesophageal function and on intra gastric and intra-oesophageal pH in patients with gastro-oesophageal reflux. METHODS: Fifty patients with non-complicated-gastro-oesophageal reflux disease classified according to Savary-Miller in: grade O, n=24; grade 1, n=19; grade 2, n=6; grade 3, n=1. Of these patients, 24 were Helicobacter pylori positive and 26 negative. Patients underwent, on two different days, stationary oesophageal manometry and 24-hour gastro-oesophageal pH-metry. RESULTS: No difference was observed between Helicobacter pylori infected and non-infected individuals with regard to lower oesophageal sphincter function, oesophageal peristalsis and gastrooesophageal reflux. These parameters were more impaired in individuals with erosive gastro-oesophageal reflux disease but this result was not dependent on the Helicobacter pylori status. Helicobacter pylori did not influence the pattern of gastric pH; however, considering only individuals with non-erosive gastro oesophageal reflux disease, gastric pH was significantly higher in infected individuals, who, histologically, also showed a corpus predominant gastritis. CONCLUSIONS: In patients with gastro-oesophageal reflux disease, Helicobacter pylori does not affect the oesophageal motility or the gastro-oesophageal reflux. These parameters are strictly related to the severity of gastro-oesophageal reflux disease as assessed at endoscopy. In patients with non-erosive gastro oesophageal reflux disease, a corpus predominant Helicobacter pylori gastritis could be responsible for the less severe gastro-oesophageal reflux. PMID- 11030181 TI - Antibody response to Helicobacter pylori CagA and heat-shock proteins in determining the risk of gastric cancer development. AB - OBJECTIVE: To investigate whether the systemic antibody response to Helicobacter pylori heat shock protein B can be considered, in addition to anti cytotoxin associated protein [CagA) antibody determination, a further serological marker of increased risk of gastric cancer development. METHODS: A total of 98 Giemsa positive Helicobacter pylori patients (28 with gastric cancer, 30 with duodenal ulcer and 40 with nonulcer dyspepsia) were studied. Serum samples obtained from all patients were tested for IgG antibodies to CagA (116 kDa), VacA [89kDa) and heat skock protein B (54 kDa) antigens of Helicobacter pylori by the Western blot technique. RESULTS: 26/28 patients [(92.9% with gastric carcinoma, 29/30 patients [96.7%) with duodenal ulcer and 30/40 patients (75.0%) with non-ulcer dyspepsia were seropositive for CagA protein. The prevalence of serum IgG antibody to CagA in the cancer patients was not significantly higher than in duodenal ulcer and non-ulcer dyspepsia patients. The prevalence of antibodies to VacA was not significantly different between gastric carcinoma and non-ulcer dyspepsia patients. In contrast the prevalence of systemic antibodies to heat skock protein B was significantly higher in gastric cancer patients (78.6%) than in duodenal ulcer (36.7%, p=0.002) or nonulcer dyspepsia patients (52.5%, p=0.029). CONCLUSIONS: The detection of antibodies to heat shock protein B is proposed as an additional test which, in association with the determination of serum antibodies to CagA, could help in determining the risk of developing severe gastroduodenal disease, and gastric cancer, in particular. PMID- 11030182 TI - Biological marker for diseases sustained by Helicobacter pylori infection. PMID- 11030183 TI - Mortality trends of malignant tumours of digestive organs in Belgrade, Yugoslavia, 1975-1997. AB - AIM: To analyse mortality trends of malignant digestive tract tumours. MATERIAL AND METHODS: Population of Belgrade (Yugoslavia) in the period 1975-1997. Mortality rates were standardized by direct method using world population as the standard. For time series, exponential trends were calculated by the use of three year moving average rates. RESULTS: In males, the highest mortality rate was for stomach cancer: 14. 6 per 100,000 (average for the period 1975-1997), followed by cancer of liver and cancer of colon: 8.4/100,000, cancer of rectum: 8.1/100,000, cancer of pancreas: 7.3/100,000, oesophageal cancer: 2.8/100,000 and gallbladder cancer: 2.0/100,000. In males, upward mortality trends for carcinomas of colon, rectum, pancreas, oesophagus, gallbladder and bile ducts were observed. The mortality rates series for stomach cancer and liver cancer did not fit any usual trend function. In females, the highest mortality rate was also for stomach cancer 7.7 per 100,000, then for cancer of colon: 6.0/100,000, cancer of rectum: 5.3/100,000, cancer of liver: 4.4/100,000, cancer of pancreas: 4.4/100,000, gallbladder cancer 3.4/100,000 and oesophageal cancer: 0.8/100,000. In females, upward mortality trends were observed for colon and rectal cancer, cancer of pancreas, and gallbladder and bile duct cancer. Downward mortality trends were present for stomach cancer and liver cancer. Mortality rates series for oesophageal cancer did not fit any usual trend function. CONCLUSIONS: In the majority of digestive tracts cancers, an upward mortality trend was observed which is in keeping with the mortality trends of these malignant tumours in many other countries. PMID- 11030184 TI - Left ventricular volumes in liver cirrhosis. AB - BACKGROUND: Patients with alcoholic cirrhosis have left ventricular dimensions similar to controls. Few data have been reported in patients with cirrhosis of viral origin. AIM: To assess left ventricular dimensions in patients with pure viral cirrhosis. PATIENTS AND METHODS: Thirty patients with virus-related cirrhosis, 23 patients with alcoholic cirrhosis and 12 healthy controls were submitted to measurement of left ventricular volumes, cardiac output, mean arterial pressure and total peripheral resistance. RESULTS: Patients with cirrhosis showed a similar increase in cardiac index and heart rate and reduction of mean arterial pressure and peripheral vascular resistance in comparison to controls, irrespective of the aetiology. Left ventricular end systolic volume index was lower (p<0.01) and ejection fraction higher (p<0.01) in virus-related cirrhotic patients [mean +/- SD, respectively 12.4+/-4.1 ml/sqm and 77.9%) in comparison both to controls (21.5+/-6.3 ml/sqm and 66.8%) and alcoholics (20.6+/ 7.0 ml/sqm and 68.8%). End diastolic volume index was not significantly different between the three groups. CONCLUSIONS: Our findings indicate smaller left ventricular volumes and higher ejection fraction in pure virus-related cirrhosis than in alcoholic cirrhosis and controls. Since peripheral haemodynamics proved similar in virus- and alcohol-related cirrhosis, a subclinical alcohol cardiomyopathy may be hypothesised to account for the absence of such left ventricular pattern in alcoholic patients. PMID- 11030185 TI - Reduced quality of life in patients with chronic hepatitis C: effects of interferon treatment. AB - BACKGROUND: Quality of life is an area of increasing interest in hepatology. Studies, so far, have assessed quality of life in patients with chronic virus C related hepatitis in relation to antiviral therapy by means of generic questionnaires. AIM: To measure quality of life in chronic hepatitis patients without cirrhosis by means of the Nottingham Health Profile questionnaire, a measure of "distress" in comparison with the Medical Outcome Survey SF-36, an index of well-being. PATIENTS: A series of 126 outpatients with chronic hepatitis; 37 on and 89 not on active interferon treatment. METHODS: The two questionnaires were used in random order. Clinical and laboratory data were also collected. The final score of any domain of the two questionnaires, for any individual patient, was compared to age-adjusted normal values obtained in 2 random samples of Italian population. RESULTS: Patients showed a significant modification of 3 domains of Nottingham Health Profile (Energy, Social Isolation and Physical Mobility) and 6 domains of SF-36. In relation to interferon treatment, the Nottingham Health Profile questionnaire was able to detect differences in Energy, Physical Mobility and Pain, which were modified only in treated patients. SF-36 did not show any differences in relation to treatment. In addition, the Nottingham Health Profile demonstrated that treated patients had a lower prevalence of concern for family life, possibly due to expectations of treatment itself. CONCLUSIONS: Active interferon treatment causes considerable distress in chronic hepatitis C patients, adding to the perceived change in health status caused by liver disease. PMID- 11030186 TI - Comparison of two enteric coated microsphere preparations in the treatment of pancreatic exocrine insufficiency caused by cystic fibrosis. AB - BACKGROUND: Pancreatic exocrine insufficiency is a common condition in patients with cystic fibrosis. Large amounts of pancreatic enzyme supplements are required to reduce malabsorption but patient compliance is not always optimal. AIMS: To compare patients' preference and the efficacy of two enteric coated microsphere preparations in patients with cystic fibrosis. PATIENTS: Patients with pancreatic exocrine insufficiency due to cystic fibrosis. METHODS: Patients were assigned to the crossover treatment with Creon or Pancrease for 1 week and then to the alternative treatment. Patients had to follow a fixed diet (at least 2 g fat/kg) and had to assume 1000 units lipase/g fat. The evaluation parameters were: patients' preference, acceptance of therapy, stool fat excretion, stool weight, gastrointestinal symptoms, and tolerance. RESULTS AND CONCLUSIONS: Of the 33/60 patients who expressed a preference for one of the two treatments, 30 preferred Creon while only 3 patients preferred Pancrease (p<0.001). No difference between the two treatments was observed regarding stool characteristics, gastrointestinal symptoms and tolerance. The mean number of capsules taken daily was reduced by 35% with Creon. The results of this study showed a preference in favour of Creon probably due to the reduction of daily capsule intake of 35%, supporting digestion as well as Pancrease. PMID- 11030188 TI - Incidence of post-endoscopic retrograde cholangiopancreatography/sphincterotomy pancreatitis depends upon definition criteria. PMID- 11030187 TI - Incidence of post-endoscopic retrograde-cholangiopancreatography/sphincterotomy pancreatitis depends upon definition criteria. AB - BACKGROUND: The reported incidence of post-endoscopic retrograde cholangiopancreatography/sphincterotomy pancreatitis ranges between 1.3% and 12.8%. This may likely reflect different definitions of pancreatitis and methods of data collection, rather than differences in patient populations, indications and endoscopic expertise. AIMS: The present study evaluated the incidence of post endoscopic retrograde-cholangiopancreatography/sphincterotomy pancreatitis using different definition criteria and different data collection methods. PATIENTS: The 24-hour clinical and enzymatic course of 1185 procedures was recorded. METHODS: Pancreatic-like pain and hyperamylasaemia were evaluated either 6 to 8 hours or 24 hours after the procedure; computed tomography scan was performed in those patients with 24-hour pancreatic pain associated with hyperamylasaemia more than three times the upper normal limit. Results. Computed tomography scan findings consistent with pancreatitis were observed in 1.9% of cases, only among those patients with 24-hour pancreatic-like pain and hyperamylasaemia over five times the upper normal limit. The 6-8-hour and 24-hour pancreatic-like pain was associated with serum amylase levels at least three times higher in 11.7% and 6.6% and five times higher or more in 7.4% and 5.1%, respectively; 6-8 and 24 hour hyperamylasaemia higher than five times the upper normal limit, irrespective of pancreatic-like pain, was reported in 8.3% and in 6.9% of cases. No patients with serum amylase values lower than three times the upper normal limit had clinical symptoms. CONCLUSIONS: The incidence of post-procedure pancreatitis ranged from 1.9% to 11.7% depending on the definition criteria adopted. PMID- 11030189 TI - Lymphocytic gastritis and protein-losing gastropathy. AB - Lymphocytic gastritis is a histopathological entity of unknown aetiology which is characterized by dense surface and foveolar epithelial T-cell infiltration. We report here an uncommon clinical presentation in a young female presenting with unexplained recurrent weight loss and peripheral oedema. Endoscopic and histological features before and after successful therapy with omeprazole are described. PMID- 11030190 TI - The cause of colorectal cancer. PMID- 11030191 TI - Virus, liver and autoimmunity. AB - The immune system's ability to distinguish self from nonself is essential for both host defence against microbial antigens and protection of self-antigens from autoimmune destruction. Such discrimination is complicated by extensive structural homology shared between micro-organisms and self-antigens, a condition known as molecular mimicry. Molecular mimicry provides the foundation for an immune response directed against an exogenous agent such as a virus to cross react with mimicked host self sequences, leading to autoimmunity, and in some cases, tissue injury and autoimmune disease. In this review we analyse studies investigating the role of molecular mimicry and cross-reactive immunity in liver related autoimmunity. PMID- 11030192 TI - Standards for non-invasive methods for gastrointestinal motility: scintigraphy. A position statement from the Gruppo Italiano di Studio Motilita Apparato Digerente (GISMAD). AB - Scintigraphic techniques are well established methods to assess gastrointestinal transit patterns. The main advantages are represented by the possibility to evaluate, in a relatively non-invasive manner, the transit time of specific components of physiological meals as well as of some digestive secretions. These techniques are generally well accepted by the patients and results are largely not operator dependent. Their main limitation is the use of a radioisotope, that prevents repeated applications in all subjects and immediate exclusion of women of childbearing potential. Due to the high costs of these tests and the limited availability of Nuclear Medicine Departments, scintigraphic techniques should be reserved for well-selected patients. A review is made of the technical requirements and indications for scintigraphic tests currently available for clinical purposes (gastric emptying study, oesophageal transit time, evaluation of gastro-oesophageal reflux, intestinal and colonic transit time, evaluation of biliary motility and of duodenogastric reflux). PMID- 11030193 TI - Antigliadin and antiendomysial antibodies in children with Down's syndrome. PMID- 11030194 TI - Acute eosinophilic pneumonia following cigarette smoking. PMID- 11030195 TI - Minimally differentiated erythroleukemia: recognition of erythroid precursors and progenitors. PMID- 11030196 TI - Clinical features and pathogenesis of catastrophic antiphospholipid syndrome. PMID- 11030197 TI - Treatment of hyperlipidemia. AB - In the treatment of hyperlipidemia, when to begin and end therapy is important. In recent years, potent anti-hyperlipidemia drugs have been widely used, and the results of many intervention trials have shown that combinations of diet, exercise and drug therapies are effective for the primary and secondary prevention of coronary heart disease. The present paper summarizes these trials; introduces the therapy guidelines for adult hyperlipidemia established by Japan Atherosclerosis Society in 1997; and discusses the drugs for hyperlipidemia. PMID- 11030198 TI - Treatment of hypertension in chronic renal insufficiency. AB - Hypertension is the most common complication of chronic renal insufficiency (CRI) and it accelerates the rate of progression of most forms of CRI. Several large clinical trials have clearly demonstrated the efficacy of antihypertensive treatment for prevention of progression of renal failure. Angiotensin-converting enzyme (ACE) inhibitors may have therapeutic advantages. However, large scale trials include a variety of chronic renal diseases, and the origins of renal diseases are quite diverse. Moreover, the differences in sex, age, race have not been considered. Based on these trials, it was concluded that blood pressure control is the most effective means for the prevention of deterioration of renal dysfunction. Consistent with this view, our recent preliminary data provide evidence that aggressive blood pressure control using self-reported blood pressure values measured by a home blood pressure device is promising to arrest the progression of renal failure in Japanese patients with CRI. PMID- 11030199 TI - Comparison of tests for fecal lactoferrin and fecal occult blood for colorectal diseases: a prospective pilot study. AB - OBJECTIVE: This prospective pilot study was conducted to compare the usefulness of measuring fecal lactoferrin (Lf) to that of fecal occult blood (FOB) test for detection of colorectal diseases. PATIENTS AND METHODS: The subjects were 351 patients who underwent colonoscopy. A fecal sample was obtained on the day before colonoscopy. Fecal Lf was measured by enzyme-linked immunosorbent assay. The FOB test was performed by combined assay (latex agglutination) of hemoglobin and transferrin. RESULTS: The specificities of the fecal Lf and FOB tests were the same (88.7%). For patients with colorectal cancer (13), colorectal polyp (69), ulcerative colitis (18), Crohn's disease (13), non-specific colitis (8), internal hemorrhoids (60), colon diverticulum (27), and miscellaneous diseases of the colon (10), the rates of positivity for fecal Lf were 7/13, 14/69, 12/18, 7/13, 4/8, 22/60, 8/27, and 6/10, respectively. The corresponding rates for FOB were 8/13, 12/69, 11/18, 4/13, 4/8, 9/60, 2/27, and 1/10. For patients with internal hemorrhoids, the rate of positivity for fecal Lf was significantly higher than that for FOB. In other disease groups, there was no significant difference in the rate of positivity between fecal Lf and FOB. CONCLUSION: These findings suggest that measurement of fecal Lf is as useful as FOB in detecting colorectal diseases. PMID- 11030200 TI - Evaluation of a new care system provided to diabetic patients in the outpatient clinic. AB - OBJECTIVE: Evaluation of metabolic states and chronic complications is essential for maintaining a high quality of care for diabetic patients. We have assessed the quality of care in routine outpatient clinics for diabetic subjects in our university hospital, and compared with those in a newly introduced standardized clinic to evaluate the new care system. METHODS: The quality of care was assessed by the chart review in 1995, and compared with those from 1996-1997 in the "Diabetes Follow-up Clinic" which is systematically designed for the standardized care. PATIENTS: The subjects were recruited among 860 patients who visited the outpatient clinic in July and August of 1995 with a diagnosis of diabetes or glucose intolerance. Six hundred seventy-two patients whose follow-up period had been more than 6 months with clinically diagnosed diabetes were used for the analysis. RESULTS: Laboratory tests such as determination of HbA1c, and serum levels of lipids and creatinine were performed in more than 90% of the patients in the routine outpatient clinics. However, ophthalmology referral, 24-hour urine collection for the determination of creatinine clearance and albumin excretion, and electrocardiograms were not well performed and were incompletely documented (40-60% of the patients within a previous year and 70-80% in the last 2 years). In the standardized "Diabetes Follow-up Clinic", only four out of 555 diabetic patients failed to collect their 24-hour urine, and all participants had ankle blood pressure measurements, nerve conduction study, and nylon monofilament tests, etc. Furthermore, more than 95% of the patients had funduscopic examinations by ophthalmologists as well as records of electrocardiogram. CONCLUSION: Introduction of the standardized "Diabetes Follow-up Clinic" may be one of the choices for increasing the quality of outpatient care and for the prevention of chronic diabetic complications. PMID- 11030201 TI - Treatment of membranoproliferative glomerulonephritis associated with hepatitis C virus infection. Niigata Research Group of Glomerulonephritis and Nephrotic Syndrome. AB - OBJECTIVE: Interferon has been used as a new therapeutic agent for glomerulonephritis since a manifest relationship between membranoproliferative glomerulonephritis (MPGN) and hepatitis C virus (HCV) infection was documented. However, several side effects and rebound phenomenon have been significant problems. We retrospectively evaluated the therapeutic effect and safety of the standard treatment with steroids and/or immunosuppressive agents for MPGN patients with an HCV infection. METHODS: Remission and renal survival rates as well as clinical and histological data were compared between MPGN groups with or without an HCV infection. In addition, the hepatic function was followed-up after the treatment. PATIENTS: The subjects were 42 biopsy proven MPGN patients. Seven were positive for an HCV infection. Secondary causes of MPGN excluding an HCV infection were omitted. Most patients were treated with steroids and/or immunosuppressive agents. RESULTS: The mean age of the MPGN patients with an HCV infection was significantly higher than that of those without an HCV infection. The renal function and the interstitial change of the former group were significantly worse than those of the latter. Nevertheless, remission and renal survival rates were not significantly different between the two groups. None in the HCV positive MPGN group showed an impairment of hepatic function during the clinical course. However, 2 subjects died from severe pneumonia during the treatment. CONCLUSION: The standard treatment with steroids and/or immunosuppressive agents did not reveal a statistical difference in the therapeutic efficacy between MPGN patients with or without an HCV infection. However, some in the former group showed a poor prognosis. PMID- 11030202 TI - Duration of systemic corticosteroids in the treatment of asthma exacerbation; a randomized study. AB - OBJECTIVE: To determine an appropriate duration for a short course of oral steroids in cases of asthma exacerbations. SETTING: A 1,000 bed city hospital in Kobe, Japan. PATIENTS: Patients with asthma exacerbations who needed hospital admission. METHODS: Following an initial treatment with a 3-day course of intravenous methylprednisolone, patients were allocated to either a 1-week (1 W) or a 2-week (2 W) course of oral prednisolone (PSL, 0.5 mg/kg). OUTCOME MEASURES: Peak expiratory flow rate (PEF) and rate of unscheduled hospital visits and readmission. RESULTS: Twenty patients were enrolled (10 in 1 W, 10 in 2 W). Mean PEF just before starting oral PSL in 1 W and 2 W were 51 and 58% of each patient's best value. PEF was significantly improved and to a similar degree over the course of time in both 1 W and 2 W. The frequency of unexpected hospital visits during a 3-month period after discharge was similar (2 in 1 W and 2 in 2 W). No readmission occurred during the same period. CONCLUSIONS: Because both the 1-week and the 2-week course of oral PSL were equally effective in the treatment of asthma exacerbations, 1 week may be appropriate as the maximum duration of a short rescue course of oral steroids. PMID- 11030203 TI - Applicability of Hyland's Living with Asthma Questionnaire for Japanese asthmatic patients. AB - OBJECTIVE: To assess the applicability of Hyland's Living with Asthma Questionnaire (LWAQ, 1991), one of the international health-related quality of life scales, for Japanese asthmatic patients with reference to its reproducibility and validity. SUBJECTS AND METHODS: The LWAQ was given to randomly selected asthmatic patients on two occasions separated by a 12-week interval. RESULTS: The mean scale score in the first study (n=304) was 1.83 (range, 1.14-2.77) and logarithmic values of the scores approached normal distribution. The scale scores in the first and second (n=158) studies were well correlated (r=0.81), however, the mean score decreased (0.08) significantly. The questions were further separated into 11 domains. The sex-domain was notable for a low response rate (68%), and scale scores in the sleep-, colds- and sex-domains in the first study varied considerably from those of the other domains. Frequency distributions of scores in the five constructs (Hyland 1996) were not normal and, with the exception of the colds construct, the relations among the remaining four constructs were similar to those previously reported (Hyland 1996). CONCLUSION: Analysis using the mean scale score, domain and construct in the LWAQ is applicable to Japanese asthmatic patients. PMID- 11030204 TI - Long-term (14 years) effect of LDL apheresis on obstructive changes in aortocoronary saphenous-vein bypass grafts in a case of heterozygous familial hypercholesterolemia with the LDL receptor proline664 to leucine mutation. AB - A 61-year-old Japanese woman with heterozygous familial hypercholesterolemia (FH), type 2 diabetes mellitus and coronary artery disease underwent coronary artery bypass grafting (CABG) utilizing a saphenous vein graft at the age of 46, in June 1984, 6 months before low density lipoprotein (LDL) apheresis was started. She had received LDL apheresis every two weeks, along with combined drug treatment since the age of 47 (December 1984). She had bilateral xanthelasma and Achilles tendon xanthomas. Her fasting baseline serum total cholesterol and triglyceride level were 464 mg/dl and 57 mg/dl, respectively at the age of 47 when she visited our hospital for the first time. Analysis of the genomic DNA from the patient revealed heterozygous amino acid substitution of Leu for Pro664 in the LDL receptor gene. She was diagnosed as type 2 diabetes mellitus at the age of 53. Combined treatment in the steady state yielded a pretreatment LDL cholesterol level of 230+/-14 mg/dl and a posttreatment level of 57+/-7.6. All grafts were widely patent after as long as 14 years since CABG, suggesting that LDL apheresis combined with drug therapy is highly effective in preventing the occlusion of bypass grafts in a patient with heterozygous FH and type 2 diabetes mellitus. PMID- 11030205 TI - Hypercalcemia induced with the plasma levels of parathyroid hormone-related peptide in multiple myeloma. AB - A 69-year-old man visited our department of neurology with symptoms of paresthesia on the lower extremities and lumbago. Biochemical examination of serum samples showed hypercalcemia (serum concentration 15.6 mg/dl). The levels of intact parathyroid hormone (i-PTH) and 1,25-dihydroxyvitamin D were suppressed, whereas parathyroid hormone-related peptide (PTHrP) was elevated up to 5.4 pM (normal range: below 0.6 pM). Additionally, bone survey revealed a punched-out lesion in radiological examinations of the skull. Bone marrow aspiration demonstrated many atypical plasma cells suggesting multiple myeloma. Nephrogenous cyclic adenosine monophosphate (cAMP), urinary deoxypyridinoline, plasma interleukin 6 (IL-6) and transforming growth factor beta (TGF beta) concentrations were elevated, whereas % of renal tubular reabsorption of phosphate (%TRP) was decreased. The immunohistochemical results demonstrated the expression of PTHrP in atypical plasma cells. These data indicated that hypercalcemia complicating multiple myeloma causes an elevation of renal calcium reabsorption and an increase of bone resorption mediated by PTHrP action. PMID- 11030206 TI - Undefined complications of parathyroid adenoma, parathyroid hyperplasia (primary hyperparathyroidism), thyroid follicular adenoma, thyroid papillary carcinoma, temporal astrocytoma, cerebellar meningioma, and hemangioma of external auditory meatus and oral papilloma. AB - A 59-year-old woman who had parathyroid adenoma, parathyroid hyperplasia, thyroid follicular adenoma, thyroid papillary carcinoma, astrocytoma of the right temporal lobe, cerebellar meningioma, capillary hemangioma of the left external auditory meatus and papilloma of the left upper gingiva is reported. Dynamic magnetic resonance imaging, computed tomography with contrast-enhancement and gastrofiberscopy revealed no remarkable findings in the pituitary, pancreas, adrenals, stomach or duodenum. Similar lesions were not found in any family members. Defect of the causative genes of multiple endocrine neoplasia types I and IIa, MENIN and RET was not detected. Further follow-up of this patient and family members is needed. PMID- 11030207 TI - Malignant pheochromocytoma lacking clinical features of catecholamine excess until the late stage. AB - A malignant pheochromocytoma is described in a 71-year-old man. Osseous metastases became manifest 12 years after successful removal of the primary tumor which originated in paraganglionic tissue near the right adrenal gland. Although the patient had no symptoms of catecholamine excess initially, hypertension, tachycardia and excessive sweating appeared several months before his death, concomitantly with a sharp increase in noradrenaline secretion due to an accelerated growth of metastatic tumors. Since there is no histologic criterion of malignancy in this neoplasm, it would be prudent to consider every case of pheochromocytoma as potentially malignant and to follow-up carefully for a long time after removal of the primary tumor. PMID- 11030208 TI - Paralytic ileus accompanied by pneumatosis cystoides intestinalis after acarbose treatment in an elderly diabetic patient with a history of heavy intake of maltitol. AB - An 87-year-old woman, diagnosed with diabetes mellitus at age 73, exhibited abdominal distention and appetite loss in March 1998. She had received acarbose as well as 5 mg per day of glibenclamide and had habitually used about 100 g of maltitol daily from 1997. She was diagnosed as having paralytic ileus accompanied by pneumatosis cystoides intestinalis (PCI). This condition subsided quickly with discontinuation of diet or cessation of acarbose and maltitol usage. The patient's condition appears to be attributable to increased gas levels produced by fermentation of disaccharides and maltitol. Decreased intestinal motility may be a result of pre-existing diabetic autonomic neuropathy and hypothyroidism. The patient's clinical course suggests that paralytic ileus and PCI should be listed as rare side effects of alpha-glucosidase inhibitors and that the drug should be used with great caution for those who consume non-digestive sugar substitutes. PMID- 11030209 TI - Acute eosinophilic pneumonia following cigarette smoking. AB - Two cases of acute eosinophilic pneumonia following cigarette smoking are analyzed for characteristic features. The first patient noted dyspnea 14 days after initiation of smoking. The second patient noted dyspnea 12 days after beginning to smoke. Both cases had characteristic features including occurrence at an age younger than 30 years; less than 1 month duration of cigarette smoking before onset of disease; and no identifiable cause of acute eosinophilic pneumonia apart from smoking. We believe that acute eosinophilic pneumonia following cigarette smoking, which has characteristic features as described above, should be considered as a distinct subtype of AEP. PMID- 11030210 TI - Two cases of atypical carcinoid of the thymus. AB - We describe two cases of atypical carcinoid of the thymus. One was an 82-year-old woman with complaints of nonproductive cough and back pain, and the other a 64 year-old woman with no symptoms. Computed tomography scans of the chest in both cases revealed a large mass in the anterior mediastinum. Multiple metastases to bone and liver were also noted in the former case. Histological examination of their tumors revealed that the tumor cells were arranged in a nested, trabecular, or pseudorosette pattern, with increased numbers of mitoses, nuclear pleomorphism, and presence of necrosis. In addition, immunohistochemically, the cells stained for neuron-specific enolase, synaptophysin and chromogranin A. Combination chemotherapy consisting of carboplatin and etoposide was performed as initial chemotherapy in the former case and as adjuvant therapy in the latter. The former patient achieved a short-term partial response. It is important to differentiate atypical carcinoid from other thymic tumors, since such tumors including thymoma have a much better prognosis than does atypical carcinoid. PMID- 11030211 TI - An unusual case of mediastinal lymphadenopathy caused by amyloidosis. AB - A 71-year-old Japanese female Nagasaki-atomic-bomb survivor was admitted for evaluation of a mediastinal mass. She was infected with human T-cell leukemia virus type I. Histological examination of the biopsy specimen, obtained thoracoscopically from the mass, revealed amyloid lymphadenopathy confirmed by Congo-red staining and electron-microscopic examination. Amyloid deposits and the serum monoclonal peak consisted of immunoglobulin lambda light chains. No atypical cells were detected in bone marrow. The case was diagnosed as an unusual amyloidosis affecting the mediastinal lymph node. No other sites were found to be affected by amyloidosis, although systemic involvement could not be conclusively ruled out. PMID- 11030212 TI - Sialyl-Tn- and neuron-specific enolase-positive minimally differentiated erythroleukemia. AB - Flow cytometric, immunochemical and molecular studies were performed on leukemic blasts from a patient with minimally differentiated erythroleukemia (AML-M6v). The blasts expressed CD36 and CD71 but not lymphoid antigens, myeloid antigens, CD41 or glycophorin A. Analysis of carbohydrate antigens showed that the blasts expressed the sialyl-Tn antigen. Immunochemistry revealed that the blasts had neuron-specific enolase (NSE). Serum sialyl-Tn and NSE levels were markedly increased. Finally, an erythroid lineage was confirmed in the presence of alpha globin messages in the blasts. Sialyl-Tn and NSE expression in leukemic blasts may be useful to identify AML-M6v. PMID- 11030213 TI - Sensory ataxic dominant neuropathy associated with polyarteritis nodosa. AB - A 68-year-old man with sensory ataxic dominant neuropathy associated with polyarteritis nodosa (PAN) had deep sensory disturbance with unsteady gait and absence of the Achilles tendon reflex. Examination revealed weight loss, elevated CRP level, negative antineutrophil cytoplasm antibodies, decreased M-wave amplitude in the peroneal motor nerve and absence of action potentials in the sural sensory nerve. Sural nerve biopsy revealed a marked loss of myelinated fibers, myelin ovoid formation and necrotizing angiitis of large epineurial arterioles. Renal biopsy revealed global and/or segmental necrotizing angiitis in glomeruli, but not in the arcuate artery. These pathological findings were distinct from those of PAN, particularly microscopic polyangiitis (MPA). Treatment with a steroid improved the deep sensory disturbance, unsteady gait, and CRP level. This case is similar to ataxic neuropathy which can arise from various disorders. This is the first report of a case of sensory ataxic dominant neuropathy associated with MPA in PAN. PMID- 11030214 TI - Alternate numbness in the upper extremities as the initial symptom of basilar migraine: an electrophysiological evaluation using EEG power topography. AB - A case of basilar migraine (BM) with alternate numbness as the initial symptom is described. The patient's chief complaint was alternate numbness in the right and left upper extremities. After angiography the patient fell into a drowsy state, followed by excitation, and finally confusion. The EEG power topography showed slow alpha, theta and delta power in the right occipital area, and alternatively in the right and left parietal area. These findings suggest that the cause of BM is not only based on a vasoconstriction mechanism, but also cortical spreading depression. BM should be suspected as a cause of sensory symptoms. PMID- 11030215 TI - Catastrophic antiphospholipid antibody syndrome in systemic lupus erythematosus: an autopsy case report of a young woman. AB - Catastrophic antiphospholipid syndrome (CAPS) is a severe variant of antiphospholipid syndrome (APS) characterized by disseminated microangiopathy that results in multiorgan failure. CAPS mainly occurs in association with systemic lupus erythematosus (SLE). Clinically, CAPS mimics disseminated SLE vasculitis, intravascular coagulation (DIC), and particularly thrombotic thrombocytopenic purpura (TTP). We describe an autopsy case of young woman with CAPS in SLE, which is difficult to differentiate from TTP secondary to SLE. PMID- 11030217 TI - Ischemic colitis probably induced by refractory constipation after voglibose administration in a patient with total gastrectomy. PMID- 11030216 TI - Methemoglobinemia induced by combined use of sodium nitrate and acetoaminophen. PMID- 11030218 TI - Use of influenza vaccine in patients with diabetes, particularly those with complications of cardiac or renal disease. PMID- 11030219 TI - Parkinsonism onset in a patient concurrently using tiapride and donepezil. PMID- 11030220 TI - Bone marrow transplantation during the 1995 Hanshin earthquake disaster. PMID- 11030221 TI - Chromosome evolution involving Robertsonian rearrangements in Xyrichthys fish (Labridae, Perciformes). AB - Three Xyrichthys fish (Labridae, Perciformes), X. pavo, X. dea, and X. twistii, were cytogenetically studied. X. pavo and X. dea had 2n = 44 chromosomes, which were all acrocentric. X. twistii had 2n = 22 chromosomes consisting of eighteen meta- and submetacentric and four acrocentric chromosomes. The cellular DNA contents of X. pavo and X. twistii measured using flow cytometry were nearly equal. These results suggest that the karyotype of X. twistii evolved by decreasing the number of chromosomes by fusion events, probably Robertsonian fusion. Cytogenetic relationships among the three species were surmized on the basis of features on the karyotypes and the NOR locations. A large gap in the chromosome number between 2n = 44 and 2n = 22 is an interesting feature related to the process of chromosome evolution. PMID- 11030222 TI - Status of phosphatase activities in the liver and kidney of rats treated with isosaline leaf and stem-bark extracts of Harungana madagascariensis (L). AB - The activities of phosphatases and other biochemical parameters were examined in rats treated with isosaline leaf and stem-bark extracts of Harungana madagascariensis (L). Results show that both the leaf and stem-bark extracts significantly increased the activities of serum and liver alkaline phosphatase, liver acid phosphatase, liver and kidney glucose-6-phosphatase, fructose-1,6 diphosphatase and glycogen in the treated rats. While the stem-bark extract significantly elevated the activities of fructose-1,6-diphosphatase and glycogen in the kidney, these biochemical parameters were not affected by treatment with the leaf extract. The activity of serum acid phosphatase was unaffected by the two extracts. The results obtained clearly show that these extracts caused a marked increase in gluconeogenesis in the liver and kidney of the treated rats. While the stem-bark extract increased gluconeogenesis in both liver and kidney, the leaf extract caused an increase in gluconeogenesis only in the liver. The increased serum alkaline phosphatase activity caused by these extracts may, aside from having other tissues contributing to it, be due to damage caused by these extracts to the hepatocytes. The extent of pathological changes as well as the implications of these findings to folklore medicine requires further investigation. PMID- 11030223 TI - Chromosomal studies on five species of the genus Leptodactylus Fitzinger, 1826 (Amphibia, Anura) using differential staining. AB - Cytogenetic studies were carried out on five species of Leptodactylus, namely L. fuscus, L. notoaktites, L. labyrinthicus, L. ocellatus, and L. podicipinus, after standard staining, Ag-NOR and C-banding as well as BrdU incorporation for three of them. The species had 2n = 22 chromosomes and two basic karyotype patterns. Chromosome 8 was a marker bearing a secondary constriction. In all species, this secondary constriction corresponded to the Ag-NOR site. The species had centromeric C-bands in all chromosomes of the complement, but some interstitial or telomeric bands seemed to differentiate some karyotypes, either at the species or the population level. In L. ocellatus, the C-banding pattern confirmed the occurrence of a heteromorphic pericentric inversion in chromosome 8 in specimens from one of the populations. The BrdU incorporation technique showed no detectable difference in the replication patterns of the major bands in the chromosomes of L. notoaktites, L. labyrinthicus, and L. ocellatus. PMID- 11030224 TI - The sequestration of hydroxyl ions by CO2 in liquid water: the physiological implications and the second function of carbonic anhydrase. AB - The pH changes due to bubbling CO2 through water produced anomalies which were more readily explained by an hypothesis based on electrostatic attractions between the molecules. The present studies have suggested that an hexagonal array of six carbon dioxide molecules could bind and sequester a hydroxyl anion. The binding energy of the complex is estimated to be comparable with that of a covalent compound and its dissociation may only occur at the water interface with air or at the water/hydrophobic protein interface in a protein cleft. The physiological importance lies in the consequential release of an equal number of free hydrogen ions (H3O+) and the disruption of the normal action of buffer systems in regulating the cytoplasmic pH. The counteraction of this sequestration reaction and the acid-base disturbances which result, form the second important function of carbonic anhydrase isoforms, the mechanisms of which are briefly discussed. PMID- 11030225 TI - Serum protein index in human malaria. AB - The serum protein levels were determined in 158 malarial patients infected with Plasmodium falciparum. Contrary to an earlier reported protein deficiency during malarial infection, the results obtained from this study showed no significant change (p > 0.05) in serum protein levels when compared with the controls. The significance of the results related to excessive protein catabolism in fever is discussed. PMID- 11030226 TI - Verocytotoxin-producing Escherichia coli O157 on a farm open to the public: outbreak investigation and longitudinal bacteriological study. AB - Verocytotoxin-producing Escherichia coli (VTEC) O157 phage type 2 (PT2) was isolated from three calves and two goats on a farm open to the public. Phenotypic and DNA-based typing showed that the strains were identical or very closely related to those obtained from an outbreak of VTEC O157 infection in two separate family groups who visited the farm. No VTEC O157 PT2 was isolated again from the farm during a 12-month longitudinal bacteriological study undertaken after the infected animals had been removed. However, phenotypically and genotypically indistinguishable VTEC O157 PT2/28 strains were detected in two of 474 faecal samples collected at monthly visits from 15 species of animals of various ages. The two isolates were obtained from calves from different sources sampled 146 days apart, suggesting that the infection had persisted on the farm although it was not detected in the other species. The same strain was subsequently isolated from another calf housed in the same pen as one of the infected calves. The longest period during which the organism was excreted was seven days. No VTEC O157 was isolated either from 204 replacement animals (including 138 orphan lambs and 10 calves) brought in from various sources, and sampled while they were kept in isolation for two weeks before being introduced to the farm, or from environmental samples. During the study a visitor became ill with VTEC O157 PT2. However, the isolate was distinct from those recovered from the farm and there was no evidence to suggest that the visit was the source of the infection. PMID- 11030227 TI - Intra-atrial rhabdomyoma causing chylopericardium and right-sided congestive heart failure in a dog. AB - A six-year-old Staffordshire bull terrier had a four-week history of progressive lethargy, ascites and exercise intolerance. A mass 3 cm in diameter obliterating the right atrial lumen was detected by echocardiographic examination. There were lactescent pericardial and serosanguineous pleural effusions. Histological and immunochemical examinations showed that the mass was a primary rhabdomyoma. PMID- 11030228 TI - Early diagnosis of pregnancy in sows by ultrasound evaluation of embryo development and uterine echotexture. AB - Two operators attempted to detect pregnancy ultrasonographically in 196 sows daily from 15 to 25 days after insemination; 20 unbred sows were also investigated. The probe was applied transcutaneously on the right abdominal wall near the last three mammary glands. During each examination, the embryos were visualised and their transverse and longitudinal dimensions were measured. Pregnancy was confirmed by an ultrasonographic detection of embryos five days after the first ultrasound diagnosis and finally 30 to 32 days after insemination. The accuracy of diagnosis was less than 83 per cent on days 15, 16 and 17 but improved to more than 90 per cent from day 18 onwards. The uterine echotexture was studied in seven sows at oestrus and 15, 17, 19, 21, 23 and 25 days after insemination. The echotexture was more homogeneous from days 15 to 25 after insemination than at oestrus. PMID- 11030229 TI - Diagnosis of teat stenosis in dairy cattle by real-time ultrasonography. PMID- 11030230 TI - Anovulation to a luteinising hormone surge in an aged goat with follicular cysts. PMID- 11030231 TI - Retrospective evaluation of 80 non-surviving buffaloes with diaphragmatic hernia. PMID- 11030232 TI - Hunting inquiry. PMID- 11030233 TI - Pet travel scheme. PMID- 11030234 TI - Pet travel scheme. PMID- 11030235 TI - Purchase and supply of medicines. PMID- 11030236 TI - RCVS annual retention fee. PMID- 11030237 TI - Proposals for new RCVS certificates. PMID- 11030238 TI - Domestic violence and the trauma surgeon: results of a study on knowledge and education. AB - BACKGROUND: Despite the frequency of domestic violence in trauma patients, little emphasis has been placed on this subject in the education of surgeons and emergency medicine physicians. The 1997 Advanced Trauma Life Support (ATLS) course included, for the first time, education about domestic violence. This study was done to test the hypothesis that baseline knowledge about domestic violence in trauma care providers is poor and is not improved by the 1997 ATLS course. STUDY DESIGN: A study on domestic violence was designed using attending general surgeons, general surgery and emergency medicine residents, and medical students as test subjects. An educational lecture and pre- and post-tests were developed using the Eastern Association for the Surgery of Trauma position paper on domestic violence and other peer-reviewed literature as information sources. Data collected included level of training, date ATLS course was taken, and pre- and post-test scores. Statistical analysis was performed with ANOVA, with significance attributed to p < 0.05. RESULTS: Ninety-two subjects attended the lecture and completed the pre- and post-tests. The overall mean pre-test score was 54 +/- 1. There was no difference in scores for the 1997 ATLS cohort (with domestic violence material) versus the group with earlier ATLS courses (52 +/- 2 versus 51 +/- 1). The group that had never taken ATLS scored significantly better on the pre-test than the other groups (58 +/- 2, p < 0.05). All groups had significantly increased scores on the post-test (mean 77 < 1, p < 0.001 versus pre-test). CONCLUSIONS: Baseline knowledge about domestic violence among surgeons and emergency medicine physicians was poor and was not improved by participation in the 1997 ATLS course. This study strongly supports the need for expanded domestic violence education for trauma care providers. PMID- 11030239 TI - Parameters that predict nipple involvement in breast cancer. AB - BACKGROUND: Breast conservation therapy has been shown to produce survival rates equivalent to those seen with modified radical mastectomy. Synchronous occult neoplastic involvement of the nipple may lead to incomplete excision of the tumor in patients undergoing breast conservation therapy, possibly leading to recurrence. STUDY DESIGN: The charts of 803 breast cancer patients treated between 1990 and 1995 at two teaching hospitals were retrospectively reviewed. The patients were divided into three groups: nipple-positive for malignancy (n = 54), nipple-negative for malignancy (n = 404), and nipple-not-removed (n = 345). Ten different clinical and tumor parameters including age, race, primary tumor location, histologic grade, primary tumor size, nodal involvement, TNM stage, estrogen receptor status, DNA ploidy, and S-phase were examined for the ability to predict cancerous nipple involvement. RESULTS: Overall, the rate of nipple positivity was 12%. In univariate analysis pathologic stage, tumor size, lymph node status, histologic grade, and tumor location were significant predictors of positive nipple involvement. Patients with tumors that were stage III or higher were nearly ten times (odds ratio [OR] = 9.8, 95% confidence interval [CI] = 5.5 to 17.7) more likely to have nipple involvement than patients with early-stage tumors. Patients with a tumor size of 4 cm or greater were nearly eight times (OR = 7.8, 95% CI = 4.2 to 14.5) more likely to have nipple involvement than patients with tumor size less than 4 cm. Patients with positive lymph nodes were five times (OR = 5.0, 95% CI = 2.7 to 9.1) more likely to have nipple involvement than patients with negative lymph nodes. Patients with tumors in a central location or that overlapped quadrants were nearly four times (OR = 3.8, 95% CI = 2.2 to 6.8) more likely to have nipple involvement than patients with tumors in other locations. Patients with grade 3 or undifferentiated tumors were three times (OR = 3.0, 95% CI = 1.4 to 6.4) more likely to have nipple involvement than patients with lower grade tumors. In multivariable analysis, stage > or = 3 (OR = 9.2, 95% CI = 4.2 to 20.3) central/ overlap location (OR = 4.1, 95% CI = 2.0 to 8.7) and grade 3 or undifferentiated (OR = 3.1, 95% CI = 1.3 to 7.5) were the only variables that remained significant predictors of nipple involvement. CONCLUSIONS: The decision to perform breast conservation surgical procedures with nipple preservation can be difficult, particularly in patients with larger, more centrally located tumors. The multivariable model developed in this study may be useful in predicting the risk of cancerous nipple involvement and selecting appropriate breast conservation patients for nipple preservation. PMID- 11030240 TI - The influence of esophageal length on outcomes after laparoscopic fundoplication. AB - BACKGROUND: It has been suggested that laparoscopic antireflux surgery has been associated with an increased incidence of postoperative paraesophageal hiatus herniation, and that this comes (at least in part) from not performing an esophageal lengthening procedure in patients with preoperative esophageal shortening. This study was undertaken to determine whether patients with esophageal shortening have an increased risk of reoperation after laparoscopic antireflux surgery. STUDY DESIGN: All patients who underwent a laparoscopic fundoplication between December 1991 and March 1999, and who had undergone preoperative esophageal manometry in our department were included in this study. Preoperative, operative, and followup data were collected prospectively, and original manometry recordings were reviewed to determine the length of the esophagus (the distance between the midpoints of the upper and lower esophageal sphincters). An index of esophageal length versus height was also calculated by dividing esophageal length by height. Esophageal length and the index were then compared with clinical outcomes. In addition, outcomes for the 50 patients with the shortest index was compared with outcomes of the 50 patients with the longest index. RESULTS: This study included 484 patients from an overall experience of 774 laparoscopic antireflux procedures. Postoperative followup ranged from 3 months to 5 years (median 2 years). Mean esophageal length was 23 cm (range 14 to 30 cm). There was a significant correlation between height and esophageal length (r = 0.44, p < 0.0001). Although patients with large hiatus hernias tended to have a shorter esophagus, preoperative endoscopic esophagitis grading did not influence length. Esophageal length did not influence the overall requirement for further surgical reintervention, although an analysis of esophageal length in patients who developed specific complications demonstrated that postoperative paraesophageal herniation was more likely in patients with a shorter esophagus, and reoperation for a tight esophageal hiatus was less likely in patients with a short esophagus. The incidence of paraesophageal hernia in the 50 patients with the shortest index was 8% versus 2% in the 50 patients with the longest index (p = 0.36). CONCLUSIONS: Although the overall reoperation rate after laparoscopic fundoplication was not influenced by esophageal length, this study did demonstrate an association between esophageal shortening and postoperative paraesophageal herniation. But the increased risk of this problem is small, and for this reason a case cannot be made for patients with a manometrically short esophagus to routinely undergo an esophageal lengthening procedure. PMID- 11030241 TI - Elective colonic operation and prosthetic repair of incisional hernia: does contamination contraindicate abdominal wall prosthesis use? AB - BACKGROUND: Wound infection and sepsis leading to incisional hernia development are common after emergency colonic operations. Later on, while being operated on to correct an incisional hernia, most of these patients will need colonic resection or bowel continuity reestablishment. Simultaneous treatment of incisional hernias in patients with colostomy or colonic disease remains a difficult challenge, considering the reluctance of most surgeons to treat both conditions at the same time, especially when prosthetic repair is needed. STUDY DESIGN: The aim of this study was to analyze the short-term results of patients undergoing colonic resection or bowel continuity reestablishment and simultaneous incisional hernia repair with an onlay polypropylene mesh technique. Over a period of 6 years, 20 patients were operated on for colonic problems associated with incisional hernias, including 8 Hartmanns' colostomies, 6 colostomies or ileostomies with colonic mucous fistulas, 3 postoperative colocutaneous fistulas, a paracolostomic hernia, a Chagas' megacolon, and a pseudotumoral diverticulitis. A "rule of three" statistical analysis was used to estimate the maximum risk of adverse effects, concerning mesh-related morbidity, after 1- and 2-year followup. RESULTS: A major complication occurred in a patient who developed an anastomotic leakage and secondary wound infection; the patient was treated with parenteral nutrition and antibiotics. Other complications included a minor wound infection, a seroma, and a chronic sinus. One patient died from postoperative problems unrelated to the surgical technique. The occurrence of postoperative wound infection did not prevent mesh incorporation. Followup ranging from 1 to 7 years detected no hernia recurrences; 13 patients were followed for 2 years or more. Our results suggest that risk of mesh-related morbidity does not exceed 15.8% (3 of 19) within the first year and 23.1% (3 of 13) for 2 years followup, with 95% confidence. CONCLUSIONS: We concluded that prosthetic repair of incisional hernias associated with simultaneous colonic operations was possible, allowing abdominal wall anatomy reestablishment. There is no reason to believe that abdominal wall prostheses must be avoided in contaminated operations when an adequate surgical technique is used. PMID- 11030242 TI - Acute pancreatitis after abdominal vascular surgery. AB - BACKGROUND: Retroperitoneal dissection and ischemia have been proposed as risk factors for postoperative pancreatitis. Although both are routine components of abdominal vascular operations, postoperative pancreatitis has not been adequately evaluated in vascular patients. The purpose of this study was to determine the incidence and outcomes of pancreatitis after abdominal vascular surgery. STUDY DESIGN: We collected pre-, intra-, and postoperative data on 21 patients who developed pancreatitis after abdominal vascular operations. For comparison, we studied 21 age- and gender-matched case controls undergoing identical operations during the same period. RESULTS: The incidence of pancreatitis among all patients undergoing abdominal vascular operations during the 6-year study period was 1.8%. Pancreatitis was diagnosed 9.8 +/- 8 days after operation and was associated with 3 or less Ranson signs in all 21 study subjects. The following outcomes data differed between the two groups: duration of npo (9 +/- 6 days for subjects versus 3.9 +/- 2 days for controls, p < 0.001) and need for parenteral nutrition (13 subjects versus no controls, p < 0.00 1). Although there was a trend towards longer hospitalization in the subjects (16 +/- 12 days versus 11 +/- 8 days, p = 0.08), there was no difference in complication rates between the two groups. Sixteen subjects (76%) had no complications. Three developed severe complications, two of whom died of causes unrelated to pancreatitis. One developed a pseudocyst that resolved spontaneously. Cholelithiasis was a causative factor in 2 subjects; no cause was established in the remaining 19. There was no difference in operative details between the two groups. CONCLUSIONS: These data indicate that pancreatitis is a rare and self-limited complication of abdominal vascular surgery. Our findings suggest that pancreatitis is costly and inconvenient but rarely serious after abdominal vascular operations. PMID- 11030243 TI - Survival and recurrence after hepatic resection of 386 consecutive patients with hepatocellular carcinoma. AB - BACKGROUND: Although hepatic resection is one of the most effective treatments for hepatocellular carcinoma (HCC), the longterm results of hepatic resection of this malignancy are far from satisfactory. The potential benefits of hepatectomy for patients with HCC have not been fully delineated. This study aimed to identify surgical outcomes of 386 consecutive patients with HCC undergoing hepatic resection. STUDY DESIGN: The retrospective study looked at records of 293 men and 93 women. The mean age was 63.2 years. Preoperative transarterial chemoembolizaton and portal vein embolization were performed in 138 patients (35.8%) and 8 patients (2.1%), respectively. Sixty-two patients (16.1 %) had major hepatectomy and the other 324 (83.9%) had minor hepatectomy. Thirty-seven of 386 patients (9.6%) had a noncurative operation. RESULTS: The 30-day (operative) mortality rate was 4.1%, and there were 11 additional late deaths (2.9%). Two hundred fourteen of 327 patients (65.4%) had recurrence after curative resection. Unfavorable factors for survival and recurrence were resection between 1983 and 1990, Child class B or C, cirrhosis, a high value of indocyanine green retention-15, a large amount of intraoperative blood loss, stage IV disease, positive surgical margin, vascular invasion, and postoperative complications. Preoperative transarterial chemoembolization increased the recurrence rate and showed no contribution to prognosis. Currently, 106 patients (27.5%) are alive: 7 (1.8%) after more than 10 years and 43 (11.1%) after more than 5 years. Mean and median overall survivals after operation were 38 months and 29 months, respectively. The 5-year and 10-year overall or disease-free survival rates after hepatic resection were 34.4% and 10.5% or 23.3% and 7.8%, respectively. CONCLUSIONS: The longterm survival rate after operation remains unsatisfactory mainly because of the high recurrence rate. Preoperative transarterial chemoembolization should be avoided because of a high risk of postoperative recurrence. Treatment strategies for recurrent HCC may play an important role in achieving better prognosis after operation, especially in patients with more than Child class B, cirrhosis, high values of indocyanine green retention-15, massive intraoperative blood loss, stage IV disease, positive surgical margin, vascular invasion, and postoperative complications. PMID- 11030244 TI - Liver transplantation for hepatocellular carcinoma: a proposal of a prognostic scoring system. AB - BACKGROUND: The current staging system of hepatocellular carcinoma established by the International Union Against Cancer and the American Joint Committee on Cancer does not necessarily predict the outcomes after hepatic resection or transplantation. STUDY DESIGN: Various clinical and pathologic risk factors for tumor recurrence were examined on 344 consecutive patients who received hepatic transplantation in the presence of nonfibrolamellar hepatocellular carcinoma to establish a reliable risk scoring system. RESULTS: Multivariate analysis identified three factors as independently significant poor prognosticators: 1) bilobarly distributed tumors, 2) size of the greatest tumor (2 to 5 cm and > 5 cm), and 3) vascular invasion (microscopic and macroscopic). Prognostic risk score (PRS) of each patient was calculated from the relative risks of multivariate analysis. The patients were grouped into five grades of tumor recurrence risk: grade 1: PRS = 0 to < 7.5; grade 2: PRS = 7.5 to < or = 11.0; grade 3: PRS > 11.0 to 15.0; grade 4: PRS > or = 15.0; and grade 5: positive node, metastasis, or margin. The proposed PRS system correlated extremely well with tumor-free survival after liver transplantation (100%, 61%, 40%, 5%, and 0%, from grades 1 to 5, respectively, at 5 years), but current pTNM staging did not. CONCLUSIONS: 1) Patients with grades 1 and 2 are effectively treated with liver transplantation, 2) patients with grades 4 and 5 are poor candidates for liver transplantation, and 3) patients with grade 1 do not benefit from adjuvant chemotherapy. PMID- 11030245 TI - P-selectin and chemokine response after liver ischemia and reperfusion. AB - BACKGROUND: P-selectin plays a major role in the earliest phase of polymorphonuclear neutrophil recruitment in the hepatic microvasculature after liver ischemia and reperfusion. Leukocyte cytokine chemoattractants (chemokines) cause polymorphonuclear neutrophil activation in ischemia and reperfusion injury. In this study, we examined the role of P-selectin in the production of chemokines in the liver and lung inflammatory response after 90 minutes of warm ischemia. STUDY DESIGN: Thirty-six C57BL/6 mice were subjected to partial liver ischemia for 90 minutes. Three groups of animals were included (n = 12 per group): the sham group, the ischemic control group, and the P-selectin-deficient gene targeted mice group. After 3 hours, we evaluated liver injury measurements, serum chemokines (MIP[macrophage inflammatory protein]-1alpha and MIP-2), liver and lung tissue myeloperoxidase, and liver and lung histology. Statistical analysis included ANOVA, Student-Newman-Keuls', and Kruskal-Wallis Multiple Comparison Z value tests. RESULTS: P-selectin-deficient mice showed significant decreases in liver enzyme levels (p < 0.05) and marked decreases in serum MIP-1alpha and MIP-2 chemokine determinations (p < 0.05) when compared with ischemic controls. Neutrophil infiltration was significantly ameliorated in the liver (p < 0.05) and markedly decreased in the lung, as reflected by decreased MPO levels. Improved histopathologic features in the liver and lung were observed in the P-selectin deficient mice group compared with ischemic controls. CONCLUSIONS: Our study confirms the key role of P-selectin in the pathogenesis of liver ischemia and reperfusion and the production of chemokines. P-selectin-deficient animals had improved liver function, decreased neutrophil infiltration, and decreased MIP- 1alpha and MIP-2 responses. PMID- 11030246 TI - Longterm central venous access in gynecologic cancer patients. AB - BACKGROUND: To assess the utility and safety of three different longterm indwelling intravenous catheters in patients with gynecologic malignancies. STUDY DESIGN: A retrospective review was performed of the records of all women with gynecologic malignancies who required longterm venous access catheters and ports between 1990 and 1997. RESULTS: Two hundred sixty-eight women underwent placement of 308 indwelling catheters, of which 305 were available for analysis. Of those, 68 (22%) were Hickman catheters, 162 (53%) were infusaports, and 75 (25%) were Peripheral Access System (PAS) ports. Venous access was obtained percutaneously in 152 (50%) patients and by cutdown in 153 (50%). Prophylactic anticoagulation was used with 96 catheters (31%). Catheter placement was associated with 12 (4%) immediate complications and 87 (29%) delayed complications. The average duration of a catheter in place was 5.6 months for the Hickman, 12.5 months for the infusaport, and 16.0 months for the PAS port (p < 0.001). Bacteremia was more likely to develop in patients with Hickman catheters when compared with those with infusaports and PAS ports (19% versus 6% and 5%, respectively, p = 0.002). Thrombosis was significantly less likely to develop in patients receiving prophylactic anticoagulation (11% versus 4%, p = 0.004). Overall, the complication rate was lower with cutdown versus percutaneous access (p = 0.004). There was no statistically significant difference between the frequency of complications when correlated with the stage of disease, patient age, body mass index, or type of malignancy. CONCLUSIONS: Infusaports and PAS ports were associated with a lower risk of infection and have a longer life than Hickman catheters. The cutdown approach was associated with a lower complication rate. Low-dose prophylactic anticoagulation should be given to all patients with longterm central venous catheters. PMID- 11030247 TI - Human error in hospitals and industrial accidents: current concepts. AB - Most data concerning errors and accidents are from industrial accidents and airline injuries. General Electric, Alcoa, and Motorola, among others, all have reported complex programs that resulted in a marked reduction in frequency of worker injuries. In the field of medicine, however, with the outstanding exception of anesthesiology, there is a paucity of information, most reports referring to the 1984 Harvard-New York State Study, more than 16 years ago. This scarcity of information indicates the complexity of the problem. It seems very unlikely that simple exhortation or additional regulations will help because the problem lies principally in the multiple human-machine interfaces that constitute modern medical care. The absence of success stories also indicates that the best methods have to be learned by experience. A liaison with industry should be helpful, although the varieties of human illness are far different from a standardized manufacturing process. Concurrent with the studies of industrial and nuclear accidents, cognitive psychologists have intensively studied how the brain stores and retrieves information. Several concepts have emerged. First, errors are not character defects to be treated by the classic approach of discipline and education, but are byproducts of normal thinking that occur frequently. Second, major accidents are rarely causedby a single error; instead, they are often a combination of chronic system errors, termed latent errors. Identifying and correcting these latent errors should be the principal focus for corrective planning rather than searching for an individual culprit. This nonpunitive concept of errors is a key basis for an effective reporting system, brilliantly demonstrated in aviation with the ASRS system developed more than 25 years ago. The ASRS currently receives more than 30,000 reports annually and is credited with the remarkable increase in safety of airplane travel. Adverse drug events constitute about 25% of hospital errors. In the future, the combination of new drugs and a vast amount of new information will additionally increase the possibilities for error. Two major advances in recent years have been computerization and active participation of the pharmacist with dispensing medications. Further investigation of hospital errors should concentrate primarily on latent system errors. Significant system changes will require broad staff participation throughout the hospital. This, in turn, should foster development of an institutional safety culture, rather than the popular attitude that patient safety responsibility is concentrated in the Quality Assurance-Risk Management division. Quality of service and patient safety are closely intertwined. PMID- 11030248 TI - A centennial history of the Chicago surgical society. PMID- 11030249 TI - The American College of Surgeons and the Royal College of Surgeons of England: eighty years of friendship. PMID- 11030250 TI - Diagnosis and management of renal trauma: past, present, and future. PMID- 11030251 TI - Therapeutically accessible clinical cardiac states. PMID- 11030252 TI - Aganglionic megacolon in a young adult. PMID- 11030253 TI - Lipoma of the small bowel. PMID- 11030254 TI - Bag lavage: a closed method for pulse lavage irrigation. PMID- 11030255 TI - Portal mesenteric shunting for reconstruction of the visceral venous system. PMID- 11030256 TI - Incisional endometriosis. PMID- 11030257 TI - Changing emphases in sexuality education in U.S. public secondary schools, 1988 1999. AB - CONTEXT: Since the late 1980s, both the political context surrounding sexuality education and actual teaching approaches have changed considerably. However, little current national information has been available on the content of sexuality education to allow in-depth understanding of the breadth of these changes and their impact on current teaching. METHODS: In 1999, a nationally representative survey collected data from 3,754 teachers in grades 7-12 in the five specialties most often responsible for sexuality education. Results from those teachers and from the subset of 1,767 who actually taught sexuality education are compared with the findings from a comparable national survey conducted in 1988. RESULTS: In 1999, 93% of all respondents reported that sexuality education was taught in their school at some point in grades 7-12; sexuality education covered a broad number of topics, including sexually transmitted diseases (STDs), abstinence, birth control, abortion and sexual orientation. Some topics--how HIV is transmitted, STDs, abstinence, how to resist peer pressure to have intercourse and the correct way to use a condom--were taught at lowergrades in 1999 than in 1988. In 1999, 23% of secondary school sexuality education teachers taught abstinence as the only way of preventing pregnancy and STDs, compared with 2% who did so in 1988. Teachers surveyed in 1999 were more likely than those in 1988 to cite abstinence as the most important message they wished to convey (41% vs. 25%). In addition, steep declines occurred between 1988 and 1999, overall and across grade levels, in the percentage of teachers who supported teaching about birth control, abortion and sexual orientation, as well as in the percentage actually covering those topics. However, 39% of 1999 respondents who presented abstinence as the only option also told students that both birth control and the condom can be effective. CONCLUSIONS: Sexuality education in secondary public schools is increasingly focused on abstinence and is less likely to present students with comprehensive teaching that includes necessary information on topics such as birth control, abortion and sexual orientation. Because of this, and in spite of some abstinence instruction that also covers birth control and condoms as effective methods of prevention, many students are not receiving accurate information on topics their teachers feel they need. PMID- 11030258 TI - Sexuality education in fifth and sixth grades in U.S. public schools, 1999. AB - CONTEXT: While policymakers, educators and parents recognize the need for family life and sexuality education during children's formative years and before adolescence, there is little nationally representative information on the timing and content of such instruction in elementary schools. METHODS: In 1999, data were gathered from 1, 789 fifth- and sixth-grade teachers as part of a nationally representative survey of 5,543 public school teachers in grades 5-12. Based on the responses of 617 fifth- and sixth-grade teachers who said they teach sexuality education, analyses were carried out on the topics and skills sexuality education teachers taught, the grades in which they taught them, their teaching approaches, the pressures they experienced, whether they received support from parents, the community and school administrators, and their needs. RESULTS: Seventy-two percent of fifth- and sixth-grade teachers report that sexuality education is taught in their schools at one or both grades. Fifty-six percent of teachers say that the subject is taught in grade five and 64% in grade six. More than 75% of teachers who teach sexuality education in these grades cover puberty, HIV and AIDS transmission and issues such as how alcohol and drugs affect behavior and how to stick with a decision. However, when schools that do not provide sexuality education are taken into account, even most of these topics are taught in only a little more than half of fifth- and sixth-grade classrooms. All other topics are much less likely to be covered. Teaching of all topics is less prevalent at these grades than teachers think it should be. Gaps between what teachers say they are teaching and teachers' recommendations for what should be taught and by what grade are especially large for such topics as sexual abuse, sexual orientation, abortion, birth control and condom use for STD prevention. A substantial proportion of teachers recommend that these topics be taught at grade six or earlier. More than half (57%) of fifth- and sixth-grade sexuality education teachers cover the topic of abstinence from intercourse--17% as the only option for protection against pregnancy and STDs and 40% as the best alternative or one option for such protection. Forty-six percent of teachers report that one of their top three problems in teaching sexuality education is pressure, whether from the community, parents or school administrators. More than 40% of teachers report a need for some type of assistance with materials, factual information or teaching strategies. CONCLUSIONS: A large proportion of schools are doing little to prepare students in grades five and six for puberty, much less for dealing with pressures and decisions regarding sexual activity Sexuality education teachers often feel unsupported by the community, parents or school administrators. PMID- 11030259 TI - Adolescents' reports of reproductive health education, 1988 and 1995. AB - CONTEXT: Reproductive health education is a key strategy for promoting safe sexual behavior among teenagers. In the last decade, new initiatives in response to AIDS and growing interest in abstinence education may have changed the prevalence, content or timing of the reproductive health education provided by schools and parents. METHODS: Formal reproductive health education and communication with parents about reproductive health among males aged 15-19 were analyzed using data from the 1988 and 1995 National Surveys of Adolescent Males. Young men's reports of formal instruction were compared with reports by adolescent females from the 1995 National Survey of Family Growth. RESULTS: Between 1988 and 1995, formal reproductive health education became nearly universal among adolescent males: In 1988, 93% of teenage males received some formal instruction, compared with 98% in 1995. The percentage of teenage males who received instruction about AIDS increased from 73% to 97% and the proportion who received instruction about how to say no to sex increased from 58% to 75%. Adolescent males who had dropped out of school received significantly less reproductive health education than those who had stayed in school, however. In addition, the median age at initial instruction decreased from age 14 to 13. Many males did not receive instruction prior to first intercourse, with non-Hispanic blacks being significantly less likely than other males to receive education prior to first intercourse. In 1995, 54% of black males had received reproductive health education before they first had sex, compared with 68% of Hispanic males and 76% of non-Hispanic white males. A smaller share of adolescent males than females received reproductive health education, and males were less likely than females to receive instruction prior to first intercourse. CONCLUSIONS: During the last decade, many types of formal reproductive health education for adolescents expanded. Further efforts should focus on assuring access to timely, comprehensive and high-quality reproductive health education for all teenagers and reducing gaps in access related to race, gender and school attendance. PMID- 11030260 TI - Using randomized designs to evaluate client-centered programs to prevent adolescent pregnancy. AB - CONTEXT: Interventions to prevent adolescent pregnancy (primarily curriculum based programs) have not produced convincing evidence as to their success. Moreover, many evaluation approaches have been inadequate to assess program effectiveness. Therefore, rigorous evaluation of different kinds of interventions may help identify potentially effective strategies to prevent adolescent pregnancy. METHODS: An experimental design, in which clients were randomized to treatment and control groups, was used to evaluate the effects of a "client centered" approach to reducing pregnancy among high-risk young people in seven communities in Washington State. Four projects served 1,042 youth (clients aged 9 13), and three served 690 teenagers (primarily clients aged 14-17). Projects offered a wide variety of services tailored to individual clients' needs, including counseling, mentoring and advocacy. RESULTS: On average, clients in the treatment group at youth sites received 14 hours of service, and their teenage counterparts received 27 hours; controls received only 2-5 hours of service. At one youth site, clients were less likely to intend to have intercourse after the intervention than before; at another, they became less likely to intend to use substances. Clients at one teenage project reported reduced sexual behavior and improved contraceptive use after receiving services; teenagers at another site reported reduced sexual intentions and drug use, and a greater intention to use contraceptives. The programs showed no other effects on factors that place young people at risk of becoming pregnant, including their sexual values and educational aspirations, communication with their parents (measured at youth sites only), and sexual and contraceptive behavior (assessed for teenagers only). CONCLUSIONS: High-risk clients likely need considerably more intervention time and more intensive services than programs normally provide. Rigorous evaluation designs allow continued assessment that can guide program modifications to maximize effects. PMID- 11030261 TI - Long-term outcomes of an abstinence-based, small-group pregnancy prevention program in New York City schools. AB - CONTEXT: Despite drops in U.S. teenage birthrates, questions continue to arise about how best to reduce the country's adolescent birthrate. School-based programs continue to be considered one of the best ways to reach adolescents at risk of early sexual activity. METHODS: A total of 312 students completed a pretest, a posttest and a follow-up one year after the posttest: 125 who had participated in a 3-4-month-long abstinence-based small-group intervention led by trained social workers, and 187 in a comparison group that received no special services. RESULTS: There were few significant differences between the intervention and comparison groups at posttest. At the one-year follow-up, however, intervention students had significantly better scores on locus of control, their relationship with their parents and (among males only) their attitudes about the appropriateness of teenage sex. Measures of depression, self esteem, intentions to have sex, attitudes toward teenage pregnancy and various behaviors did not differ significantly between groups. By the time of the one year follow-up, there was no difference between study groups among females in the initiation of sexual intercourse. Among the males, initiation of sexual intercourse appeared to be higher in the intervention group than in the comparison group, but the difference was not statistically significant. Positive outcomes were especially limited among students who were already sexually active at the start of the study, a finding that emphasizes the difficulties of reaching adolescents who are already at high risk for pregnancy CONCLUSIONS: A small-group abstinence-based intervention focusing on mental health can have some impact on adolescents' attitudes and relationships (particularly with their parents). Long term evaluations are important for determining the effects of an intervention, as it is difficult to change adolescent risk behavior. PMID- 11030262 TI - Conducting adolescent sexuality research in schools: lessons learned. PMID- 11030263 TI - Sexuality education: our current status, and an agenda for 2010. PMID- 11030264 TI - Can the mass media be healthy sex educators? PMID- 11030265 TI - Teenagers educating teenagers about reproductive health and their rights to confidential care. PMID- 11030266 TI - No sexuality education is sexuality education. PMID- 11030267 TI - We have made progress but there is still work to be done. PMID- 11030268 TI - Penicillin allergy: high prevalence of a vague history in skin test-positive patients. PMID- 11030269 TI - Mites are ubiquitous: are mite allergens, too? PMID- 11030270 TI - T lymphocyte costimulatory molecules in host defense and immunologic diseases. AB - BACKGROUND: Costimulation is an essential component for the optimal induction of T cell-mediated immune responses. Manipulation of the costimulatory pathway with antibodies or genetically-engineered fusion proteins is an important strategy to treat immune-related diseases including allergy, asthma, transplantation and cancer. Recent advances have revealed several new costimulatory molecules, and the functional characteristics of each costimulatory pathway are now becoming clearer. LEARNING OBJECTIVES: In this review, we summarize basic outlines of the costimulatory systems in terms of molecular structure, expression kinetics and immunological function. We further discuss involvement and therapeutic manipulation of costimulation in several clinical diseases. DATA SOURCE: The MEDLINE database was used to review the literature related to costimulation. CONCLUSION: Costimulatory pathways play an essential role in the activation and regulation of T cell immune responses and the induction of T cell tolerance. Therapeutic manipulation of the costimulatory system demonstrates beneficial effects to treat immunological diseases in murine models as well as some clinical situations. PMID- 11030271 TI - Ten month old infant with wheezing for six months. PMID- 11030272 TI - Pediatricians' attitudes towards allergy: past and present attitudes of pediatricians towards allergy. AB - BACKGROUND: In 1971 we published a survey of pediatricians' knowledge, attitudes, and practices relative to the field of allergy. This current survey was conducted to determine how these attitudes and practices have changed over the past 27 years. METHODS: Our survey, consisting of 17 questions, was sent to 250 board certified pediatricians in the St. Louis area. Seven of the questions were similar to those in the 1971 survey, while the other ten were new questions which we thought were pertinent to the field of allergy currently. RESULTS: In 1971, 75% of the pediatricians felt the specialty of allergy had great or moderate importance for the practice of pediatrics compared with 99% in 1998. In 1998, the number of physicians who felt that there was at least some value to skin testing had increased from 1971, although, only 36% felt it had great value. In 1971, 23% of pediatricians performed their own skin tests compared with 3% in 1998. Nine percent indicated that they had never referred a patient to an allergist in 1971, compared with 2% in 1998. Our new questions looked at to whom respondents were more likely to refer patients with: (1) asthma (68% allergy, 27% pulmonary, 4% both), (2) urticaria (53% allergy, 41% dermatology, 2% both), (3) chronic eczema (45% allergy, 50% dermatology, 2% both), and (4) chronic sinusitis (24% allergy, 74% otolaryngology, 2% both). Older physicians placed greater importance on the field of allergy and skin testing and were more likely to refer to an allergist than physicians under the age of 40 years. CONCLUSION: Our results indicate that although there has been significant change in 27 years, allergists must be more aggressive in developing the knowledge, attitudes, and practices of physicians relative to the field of allergy. PMID- 11030273 TI - Penicillin allergy: prevalence of vague history in skin test-positive patients. AB - OBJECTIVE: Penicillin (PCN) skin testing is a reliable tool for predicting which patients can safely receive the antibiotic. Depending on the study, 7% to 76% of patients who have a history of PCN allergy have positive PCN skin tests. Many physicians approach patients who have a vague history of PCN allergy less cautiously than they approach those who have a convincing history suggestive of an IgE-mediated reaction. We reviewed the published literature to determine how many patients who had a history of PCN allergy and who were skin test-positive had a vague history of allergy. DATA SOURCES: By cross-referencing the keywords "penicillin" and "skin test," an Ovid MEDLINE search for English language studies published from 1966 to 1998 was performed. STUDY SELECTION: Studies in which history positive/skin test-positive patients were identified, and which contained documentation of the type of previous reaction in these patients, were included in the analysis. The MEDLINE search revealed 295 English language articles, of which 27 fulfilled the inclusion criteria. Three additional studies published prior to 1966 (and therefore not available through MEDLINE) also were found, bringing the total to 30. A "convincing" history was defined to be one likely to be IgE-mediated (such as anaphylaxis, urticaria, angioedema or pruritic rash). A "vague" history was one unlikely to be IgE-mediated (such as maculopapular rash, GI symptoms, or an unknown reaction). RESULTS: Overall, 347/1063, or 33%, of history positive/skin test-positive patients had a vague PCN allergy history, with a range of 0% to 70% among the 30 studies. CONCLUSION: A large proportion of patients who have PCN-specific IgE antibodies, as determined by skin testing, have vague PCN allergy histories. These results therefore, indicate that, like patients with convincing histories, patients with vague allergic histories should undergo PCN skin testing prior to PCN administration. PMID- 11030274 TI - Citrus red mite (Panonychus citri) is a common sensitizing allergen among children living around citrus orchards. AB - BACKGROUND: A recent investigation has suggested that Panonychus citri (citrus red mite) is the most common causative allergen in citrus-cultivating farmers with asthma or allergic rhinitis. OBJECTIVE: A cross-sectional survey was performed to evaluate the prevalence of asthma and sensitization to P. citri among children living around citrus orchards. METHODS: A total of 1,055 children living around citrus orchards were enrolled. There were 544 boys and 511 girls with a mean age of 12.3 (range 7 to 16) years. They were evaluated by the ISAAC questionnaire, and a skin prick test with 13 common aeroallergens including P. citri and Japanese cedar pollen. Children who recorded ever having experienced wheezing on the questionnaire were evaluated by methacholine bronchial provocation test and determined to have asthma if they showed bronchial hyperresponsiveness to methacholine simultaneously. RESULTS: The positive skin test response to one or more inhalant allergens except P. citri was 35.5%, but when including P. citri, the positive response increased to 37.7%. The most common sensitizing allergens in the order of decreasing frequency were Dermatophagoides pteronyssinus (26.6%), D. farinae (22.7%), P. citri (14.2%), cockroach (11.3%), and Japanese cedar pollen (9.7%). Among the children without sensitization to house dust mites, the sensitization rate to P. citri was significantly higher in those 13 and over than in those under 12 (5.8% versus 2.5%, P < .05). The prevalence of asthma was not different between the children with sensitization to P. citri and those without it (5.4% versus 4.0%, P > .05), although the prevalence was higher in those with sensitization to D. pteronyssinus or D. farinae than in those without sensitization to these allergens (7.9% versus 2.9%, P < .05; 9.7% versus 2.6%, P < .05). CONCLUSION: Citrus red mite is a common sensitizing allergen among children living around citrus orchards. The features of sensitization and development of asthma are different between citrus red mite and house dust mites. PMID- 11030275 TI - Nasal polyposis: clinical course during 20 years. AB - BACKGROUND: The etiology of nasal polyposis is mainly unknown although it has been connected with many clinical conditions. The long-term clinical course of nasal polyposis is largely unknown, because long-term followup studies on the recurrence of nasal polyposis have rarely been reported. OBJECTIVE: The aim of the study was to find out the clinical course of nasal polyposis over a long period of time. PATIENTS AND METHODS: Our report describes a 20-year follow-up study of 41 patients with nasal polyps. These patients had surgery for nasal polyp disease 20 years previously and they were initially grouped according to occurrence of (1) acetylsalicylic acid (ASA) intolerance, (2) atopic allergy (AT), and (3) intrinsic allergy-like disease (INTR). Patients were now re examined, sinus computed tomography (CT) scanning was made, and a biopsy from polyp or from mucosa of the middle turbinate was taken. RESULTS: Anterior rhinoscopy revealed polyps in 35 of 41 patients. Thus nasal polyposis was still active in 85% of patients after 20 years. Mucosal changes in paranasal sinuses were found in every patient. Anosmia or hyposmia was found in 61% (25/41) of the patients. Eight patients had had 11 or more surgical operations during the 20 year period. Of these, 88% (7/8) belonged to the ASA group. Bronchial asthma was found in all ASA intolerance patients (11/11), and in 36% (4/11) of AT and in 16% (3/19) of INTR patients, respectively. CONCLUSION: Because of the high recurrence tendency and insidious symptoms of nasal polyposis, patients will require followup for the rest of their lives. PMID- 11030276 TI - Asthma and rhinitis induced by exposure to raw green beans and chards. AB - BACKGROUND: Although the vast majority of IgE-mediated allergic reactions to foods occurs through ingestion, a few cases of unexpected allergic reactions to foods may occur through the exposure to airborne food allergen particles. METHODS: Case reports. Skin prick tests and serum-specific IgE (CAP-FEIA) were used to identify specific IgE antibodies. Bronchial provocation tests were performed to determine the clinical relevance of inhaled exposure to raw and cooked green beans and raw chards. After demonstrating specific reactivity to them, SDS-PAGE and immunoblotting of raw and cooked green beans were carried out to identify relevant antigens. RESULTS: Three women developed bronchial asthma and rhinitis after exposure to raw green beans, and one of them also when exposed to raw chards. All women tolerated ingestion of green beans. Patients reported multiple episodes while handling these vegetables for cooking activities. Allergy to green beans and chards was demonstrated by skin testing and serum-specific IgE. Bronchial challenge test with these allergens showed positive responses to raw, but not cooked, green beans and chards. Oral food challenges with green beans (raw and cooked) and chards were negative in all patients. In order to further characterize the allergenic components of these extracts, SDS-PAGE and electroblotting studies were also performed. Immunoblots of raw and cooked green beans extract showed two IgE-binding bands with apparent molecular weights of 41.1 and 70.6 kD. Interestingly, a 47-kD IgE-binding protein was detected only in raw green bean extracts. CONCLUSIONS: We report three patients who developed asthma and rhinitis caused by exposure to raw, but not to cooked, green beans and chards in a non-occupational environment. Only minor differences of IgE reactivity between nitrocellulose-blotted raw and boiled green bean extract were found. PMID- 11030277 TI - Body composition and growth in asthmatic children treated with inhaled steroids. AB - BACKGROUND: Prolonged treatment with inhaled steroids is recommended for long term control of asthma in children; however, it can interfere with growth and body composition. OBJECTIVE: The aim of this study is to answer the question whether 6 months treatment with inhaled steroids causes body fat accumulation and growth velocity reduction. METHODS: Hospital-based, open study of body composition [by dual-energy X-ray absorptiometry (DXA), bioelectrical impedance analysis (BIA) and skinfolds] and growth of 26 asthmatic children, treated for 6 months with inhaled steroids [budesonide (BUD) 400 microg/day (group 1) or fluticasone proprionate (FP) 200 microg/day (group 2)], sodium cromoglycate and beta2-agonist (salbutamol) compared with a control group of 16 asthmatic children treated only with sodium cromoglycate and beta2-agonist. RESULTS: On average, total and regional fat mass, adjusted for pubertal stage and gender, and growth velocity were similar in all three groups of patients and were not influenced by treatment (% mean change +/- 1 SD of fat mass during treatment in BUD 0.1 +/- 3.0%, FP -1.1 +/- 3%, and control -2.8 +/- 3.5%; ANOVA P > or = .05); however seven patients, two in group 1 (1 preschool child), three in group 2 (2 preschool children) and two in the control group (two prepubertal boys aged 8.5 and 9.5 year), during treatment, showed a growth velocity standard deviation score below the third percentile. CONCLUSION: A 6-month treatment with inhaled BUD and FP does not induce body fat accumulation; however, in a few preschool children the treatment was associated with growth velocity below the third percentile. Our results suggest the need for constant monitoring of growth in all asthmatic children on chronic treatment with inhaled steroids. Further studies devoted to the effects of inhaled steroids use in preschool children are needed. PMID- 11030278 TI - Effects of substance P and vasoactive intestinal peptide on interferon-gamma and interleukin-4 production in severe atopic dermatitis. AB - BACKGROUND: Recent studies have demonstrated that two T cell-derived lymphokines, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), play a crucial role in the pathogenesis of atopic dermatitis (AD). It is known that neuropeptides, such as substance P (SP) and vasoactive intestinal peptide (VIP), have various immunomodulatory effects. Elevated levels of these neuropeptides and increased staining of SP positive nerve fibers have been reported in AD patients. OBJECTIVE: The study was designed to examine the effects of SP and VIP on the production of IFN-gamma and IL-4. The aim of the study was to establish whether these neuropeptides acted to affect cytokine release in the peripheral blood mononuclear cells (PBMCs) of AD patients. METHODS: The effects of SP and VIP on the production of IFN-gamma and IL-4 in phytohemagglutinin stimulated PBMC cultures over a 48-hour period were analyzed by enzyme-linked immunosorbent assay in 15 AD patients. Non-atopic individuals were used as a control group. RESULTS: Base cytokine profiles of AD patients showed significantly decreased IFN-gamma and increased IL-4 when weighed against non-atopic controls. Compared with controls, SP had a significant percentage enhancing effect on both IFN-gamma and IL-4 production at concentrations of 10(-8) M and 10(-6) M, however, this IFN gamma up-regulatory effect of SP was reversed by spantide, a SP antagonist. The ratios of IFN-gamma: IL-4 production were significantly elevated in the SP treated AD group. Although VIP had no specific noticeable effects on the IFN gamma and IL-4 production. CONCLUSIONS: Our data may suggest that SP has an influence on the immunomodulation of AD patient by regulating IFN-gamma production, either directly or indirectly. Vasoactive intestinal peptide, on the other hand, has no modulatory effects on the cytokine production of AD patients. PMID- 11030279 TI - Autonomic regulation after exercise evidenced by spectral analysis of heart rate variability in asthmatic children. AB - BACKGROUND: Bronchial asthma is associated with abnormal autonomic nervous function in childhood. Exercise is one of the most common precipitating factors of acute asthmatic crises although the exact mechanism of autonomic regulation in asthmatic children after exercise is unclear. OBJECTIVE: The aim of this study was to investigate the features of autonomic regulation after exercise in asthmatic and control children. METHODS: Pulmonary function tests and heart rate variability spectral analysis were performed in 15 asthmatic children and 7 control children (age 6 to 15 years) during and after an exercise challenge. RESULTS: The maximum % fall of forced expiratory volume in 1 second (FEV1) was significantly greater (P < .01) in asthmatic subjects (9.1 +/- 5.1%) than in normal control subjects (1.0 +/- 2.5%). The high frequency band (HF) amplitude, an index of cardiac vagal tone, 5 minutes after exercise was significantly higher (P < .05) in the asthmatic subjects (14.4 +/- 7.9 msec) than in control subjects (5.9 +/- 2.6 msec). Furthermore, the difference in the HF amplitude between the control group and the exercise-induced asthma group was significant both 5 minutes (P < .01) and 10 minutes (P < .05) after challenge. There was a significant correlation (P = .565, P = .0165) between HF amplitude 5 minutes after exercise and the magnitude of the decrease in FEV1. On the other hand, no significant difference was observed in the low frequency band amplitude between the controls and the asthmatic subjects. The ratio of low frequency to high frequency power, which is suggested to correlate with cardiac sympathetic activity, did not differ between the two groups. CONCLUSION: These findings suggest that autonomic nervous activities, particularly vagal response after exercise, in asthmatic children is different from that in control children. PMID- 11030280 TI - Cannabis (hemp) positive skin tests and respiratory symptoms. AB - BACKGROUND: We have noted several patients who had rhinitis and/or asthma symptoms when exposed to Cannabis plants in the summer months. Cannabis plants are common in the Midwest. OBJECTIVES: To examine whether Cannabis might be a clinically important allergen, we determined Cannabis pollination patterns in the Omaha area for 5 years, the prevalence of skin test positivity, and the association with respiratory symptoms. METHODS: Airborne Cannabis (and other weed) pollens were collected using a Rotorod air impactor, and pollen counts were done using a standardized protocol. RESULTS: Measurable Cannabis pollen count was not recorded until the last 2 weeks of July. Peak pollination typically occurred during mid- to late-August, and comprised up to 36% of the total pollen counts. Cannabis pollen was not observed after mid-September. To determine the prevalence of skin test positivity, we added Cannabis to the multi-test routine skin test battery. Seventy-eight of 127 patients tested (61%) were skin test positive. Thirty of the 78 patients were randomly selected to determine if they had allergic rhinitis and/or asthma symptoms during the Cannabis pollination period. By history, 22 (73%) claimed respiratory symptoms in the July through September period. All 22 of these subjects were also skin test positive to weeds pollinating during the same period as Cannabis (ragweed, pigweed, cocklebur, Russian thistle, marsh elder, or kochia). CONCLUSIONS: The strong association between skin test reactivity, respiratory symptoms, and pollination period suggests that Cannabis could be a clinically important aeroallergen for certain patients and should be further studied. PMID- 11030281 TI - A 5-day course of oral desensitization to trimethoprim/sulfamethoxazole (T/S) in patients with human immunodeficiency virus type-1 infection who were previously intolerant to T/S. AB - BACKGROUND: Trimethoprim/sulfamethoxazole (T/S) is an essential drug in patients with human immunodeficiency virus type-1 (HIV-1) infection to prevent opportunistic infections. About 40% to 60% of them develop skin rash which leads to discontinue the medication. A precise mechanism of the reaction is not known. OBJECTIVE: To make the patients more tolerable to the medication and to make clear whether or not the reaction is caused by serum sulfamethoxazole-specific IgE. METHODS: We established a 5-day protocol, in which T/S was administered orally as a granular form in increasing doses beginning with 0.005 g (it contains trimethoprim 0.4 mg and sulfamethoxazole 2 mg) and doubled every 12 hours until the therapeutic dose was achieved. We tried to desensitize T/S in 17 patients with HIV-1 infection who were previously intolerant to T/S and measured the specific IgE in sera. RESULTS: Desensitization was successfully completed in 15 (88.2%) of the patients. In two patients who failed the desensitization, one was due to fever and the other was gastric irritation. During followup in a mean period of 16.6 months (range, 8 to 23 months) as of May, 1999, none has had Pneumocystis carinii pneumonia (PCP) while receiving T/S after desensitization. Sulfamethoxazole-specific IgE did not increase, indicating that it was not the major cause of skin rash due to T/S in our cases. CONCLUSION: These preliminary results show that most patients who were thought to be intolerant to T/S and had no sulfamethoxazole-specific IgE can be safely desensitized and received the drug subsequently as an effective prophylaxis for PCP. PMID- 11030282 TI - Lidocaine nebulization for treatment of asthma. PMID- 11030284 TI - A fifth amendment for the Declaration of Helsinki. PMID- 11030283 TI - Benefit of colchicine in the treatment of Schamberg's disease. PMID- 11030285 TI - Towards appropriate local surgery for patients with breast cancer. PMID- 11030286 TI - Diagnosis of adult GH deficiency. PMID- 11030287 TI - Recognition of premenstrual dysphoric disorder and its treatment. PMID- 11030288 TI - Visceral hyperalgesia revisited. PMID- 11030289 TI - Facing the biological weapons threat. PMID- 11030290 TI - Global information flow. PMID- 11030291 TI - Efficacy of selective serotonin-reuptake inhibitors in premenstrual syndrome: a systematic review. AB - BACKGROUND: Selective serotonin-reuptake inhibitors (SSRIs) are increasingly being used as first-line therapy for severe premenstrual syndrome (PMS). We undertook a meta-analysis on the efficacy of SSRIs in this disorder. METHODS: We searched medical and scientific databases, approached pharmaceutical companies, and reviewed citations of relevant articles to identify 29 studies of the use of SSRIs in PMS. 14 were excluded (no placebo group, preliminary report of included trial, or low quality). 15 randomised placebo-controlled trials were included. Information on study design, participants, drugs used and dosing regimens, outcome measures, side-effects, and sources of funding was extracted. Standardised mean differences between treatment and placebo groups were calculated to obtain an overall estimate of efficacy. The primary outcome measure was a reduction in overall PMS symptoms. FINDINGS: The primary analysis included data on 904 women (570 assigned active treatment and 435 assigned placebo, including 101 in crossover trials). The overall standardised mean difference was 1.066 (95% CI -1.381 to -0.750), which corresponds to an odds ratio of 6.91 (3.90 to 12.2) in favour of SSRIs. SSRIs were effective in treating physical and behavioural symptoms. There was no significant difference in symptom reduction between continuous and intermittent dosing or between trials funded by pharmaceutical companies and those independently funded. Withdrawal due to side effects was 2.5 times more likely in the active-treatment group than in the placebo group. INTERPRETATION: SSRIs are an effective first-line therapy for severe PMS. The safety of these drugs has been demonstrated in trials of affective disorder, and the side-effects at low doses are generally acceptable. PMID- 11030292 TI - GH-releasing hormone and GH-releasing peptide-6 for diagnostic testing in GH deficient adults. AB - BACKGROUND: The diagnosis of growth hormone (GH) deficiency in adults is based on provocative testing of GH secretion. The insulin tolerance test (ITT), currently the favoured test for this diagnosis, has been criticised for poor reproducibility and inconvenience. Since the combined administration of GH releasing hormone (GHRH) plus GH-releasing peptide-6 (GHRP-6) is the most potent stimulus of GH secretion, we did a multicentre study comparing GH peaks elicited by ITT with those elicited by the GHRH/GHRP-6 test in healthy controls and GH deficient individuals (cases). METHODS: 125 adult patients with organic pituitary disease and 125 healthy individuals were studied. All cases and controls were given GHRH 1 microg per kg bodyweight intravenously plus GHRP-6 1 microg per kg intravenously at 0 min and blood samples were obtained during a subsequent 120 min period. 27 controls and all cases had an ITT. Inclusion criteria were severe GH deficiency--ie, a GH peak after ITT of < or = 3 microg/L. Results of the GHRH/GHRP-6 test were analysed by receiver-operating characteristic curve methodology. FINDINGS: GH peaks seen after the GHRH/GHRP-6 test did not result in any side-effects and were not affected by age, sex, amount of adipose tissue, or by the GH assay system used. The GH mean peak after the GHRH/GHRP-6 test was 59.2 microg/L (SD 2.2) for controls and 4.1 microg/L (0.3) for cases, whereas after ITT the mean peak was 14.3 microg/L (1.7) and 0.5 microg/L (0.06), respectively. The differential peak responses of controls and cases was greater (p<0.001), for GHRH/GHRP-6 test than for ITT. When individually analysed GH peaks were a continuum, from 139.0 microg/L to 0.01 microg/L, with a cut-off point of 15.0 microg/L. The GHRH/GHRP-6 test performed well under the ROC curve analysis. For clinical utility, it is then proposed that values > or = 20.00 microg/L be considered normal and < or = 10.00 microg/L as GH deficient. INTERPRETATION: The GHRH/GHRP-6 test is a convenient, safe and reliable test for adult GH deficiency and is not confounded by clinical factors known to alter GH secretory patterns. An evoked GH concentration of > or = 15.0 microg/L accurately distinguishes between healthy and GH-deficient adults. PMID- 11030293 TI - Outcome after prenatal diagnosis of hypoplastic left-heart syndrome: a case series. AB - BACKGROUND: Traditionally, after prenatal diagnosis of hypoplastic left-heart syndrome (HLHS) couples have been offered termination of pregnancy or comfort care. Success of postnatal surgical options such as the Norwood procedure have been associated with survival of up to 60%. Whether survival is affected by the congenital anomaly being identified prenatally or postnatally remains uncertain. METHODS: We reviewed all cases of prenatally diagnosed HLHS referred to the Fetal Medicine Unit at Birmingham Women's Hospital over 6 years between 1994 and 1999. FINDINGS: 87 cases of HLHS were referred at a median gestational age (95% CI) of 23 (19-37) weeks. Of these, 53 (61%) chose prenatal karyotyping. The overall frequency of abnormal karyotype was found in seven of 59 cases (12%) and associated structural anomalies in 18 of 87 (21%). After counselling, 38 of 87 couples (44%) chose termination of pregnancy. Of the remaining 49 fetuses, 11 (23%) were not considered for postnatal surgery because of parental choice and they died after compassionate care. Of the 36 babies who had surgery postnatally, 12 survived (33%). We recorded a survival rate of 38% for the stage-1 Norwood procedure in the prenatally diagnosed HLHS in our centre. These data suggest that at the point of prenatal detection, the overall survival rate for fetuses with HLHS is 25% (if terminated pregnancies are excluded). INTERPRETATION: Fetal echocardiography allows early diagnosis of HLHS and gives clinicians the opportunity to triage this group dependent on prenatal findings, including karyotyping and the exclusion of other structural anomalies. These prospective data provide up-to-date information on the basis of which parents can make decisions. PMID- 11030294 TI - Relation between appropriateness of primary therapy for early-stage breast carcinoma and increased use of breast-conserving surgery. AB - BACKGROUND: Breast-conserving surgery is a more complex treatment than mastectomy, because a separate incision is needed for axillary lymph-node dissection, and postoperative radiotherapy is necessary. We postulated that adoption of this therapy into clinical practice might have led to discrepancies between the care recommended and that received. METHODS: We used records of the US national Surveillance, Epidemiology, and End Results tumour registry to study 144,759 women aged 30 years and older who underwent surgery for early-stage breast cancer between 1983 and 1995. We calculated the proportion undergoing at least the minimum appropriate primary treatment (defined, in accordance with the recommendations of a National Institutes of Health Consensus Conference in 1990, as total mastectomy with axillary node dissection or breast-conserving surgery with axillary node dissection and radiotherapy) during each 3-month period. FINDINGS: The proportion of women receiving appropriate primary therapy fell from 88% in 1983-89 to 78% by the end of 1995. This decline was observed in all subgroups of age, race, stage, and population density. Of all women in the cohort, the proportion undergoing an inappropriate form of mastectomy remained stable at about 2.7% throughout the study period. The proportion undergoing an inappropriate form of breast-conserving surgery (omission of radiotherapy, axillary node dissection, or both) increased from 10% in 1989 to 19% at the end of 1995. INTERPRETATION: Although most women undergo appropriate care, the appropriateness of care for early-stage breast cancer in the USA declined from 1990 to 1995. Because the proportion of all women who were treated by breast conserving surgery increased, and because this approach was more likely than was mastectomy to be applied inappropriately, the proportion of all women having inappropriate care increased. PMID- 11030295 TI - Contribution of central sensitisation to the development of non-cardiac chest pain. AB - BACKGROUND: Non-cardiac chest pain mimics angina pectoris but generally originates from the oesophagus. Visceral hypersensitivity may contribute, but its neurophysiological basis is unclear. We investigated whether central sensitisation, an activity-dependent amplification of sensory transfer in the central nervous system, underlies visceral pain hypersensitivity and non-cardiac chest pain. METHODS: We studied 19 healthy volunteers and seven patients with non cardiac chest pain. Acid was infused into the lower oesophagus. Sensory responses to electrical stimulation were monitored within the acid-exposed lower oesophagus, the non-exposed upper oesophagus, and the cutaneous area of pain referral, before and after the infusion. FINDINGS: In healthy volunteers, acid infusion into the lower oesophagus lowered the pain threshold in the upper oesophagus (mean decrease 18.2% [95% CI 10.4 to 26.0]; p=0.01) and on the chest wall (24.5% [10.2 to 38.7]; p=0.01). Patients with non-cardiac chest pain had a lower resting oesophageal pain threshold than healthy controls (45 [30 to 58] vs 64 [49 to 81] mA; p=0.04). In response to acid infusion, their pain threshold in the upper oesophagus fell further and for longer (mean fall in area under threshold/time curve 26.7 [11.0 to 42.3] vs 5.8 [2.8 to 8.8] units; p=0.04). INTERPRETATION: The finding of secondary viscerovisceral and viscerosomatic pain hypersensitivity suggests that central sensitisation may contribute to visceral pain disorders. The prolonged visceral pain hypersensitivity in patients with non cardiac chest pain suggests a central enhancement of sensory transfer. New therapeutic opportunities are therefore possible. PMID- 11030296 TI - A heroin user with a wobbly head. PMID- 11030297 TI - Emerging group-A rotavirus and a nosocomial outbreak of diarrhoea. AB - A P[6]G9 group-A rotavirus caused a protracted hospital outbreak of neonatal diarrhoea in The Netherlands. The outbreak lasted 5 months with 52 cases and an average attack rate of 40%, 46 cases were in an incubator section for neonates under 1 month of age. Rotavirus P161G9 was detected by RT-PCR in stool samples from the 31 cases tested. Emergence of this genotype in Europe may have implications for neonates lacking protective maternal antibodies and for the development of rotavirus vaccines. PMID- 11030298 TI - Reduced development of cerebral cortex in extremely preterm infants. AB - Most growth in cortical connections and complexity occurs after 25 weeks. The cerebral cortex of extremely preterm infants when imaged at gestational age 38-42 weeks had less cortical surface area and was less complex than in normal infants born around term (p<0.0148 and p<0.0002, respectively), despite similar term corrected cerebral tissue volumes. Since deficits acquired during critical periods of brain development may be permanent, these results suggest a neural substrate for the neurocognitive impairment that is frequent among such preterm infants. PMID- 11030299 TI - Treatment of solid tumours in children with tumour-lysate-pulsed dendritic cells. AB - Dendritic cells are potent stimulators of antigen-specific immune responses, including antitumour responses. We explored the use of tumour-lysate-pulsed dendritic cells in children with relapsed solid tumours. Dendritic cell treatment in children was feasible and apparently not toxic. The treatment was able to produce significant tumour regression in a child with metastatic fibrosarcoma. PMID- 11030300 TI - Early androgenetic alopecia as a marker of insulin resistance. AB - The previously proven association between androgenetic, alopecia and serious cardiovascular events raises a question of the common pathogenetic mechanism of these disorders. Our practice-based case-control study in men aged 19-50 years showed a strikingly increased risk of hyperinsulinaemia and insulin-resistance associated disorders such as obesity, hypertension, and dyslipidemia in men with early onset of alopecia (<35), compared with age-matched controls. This finding supports the hypothesis that early androgenetic alopecia could be a clinical marker of insulin resistance. PMID- 11030301 TI - Internal carotid-artery response to 5% carbon dioxide in women with polycystic ovaries. AB - Although polycystic ovary syndrome is associated with hypertension, hyperlipidaemia, and insulin resistance, mortality from cerebrovascular disease is not increased. We previously reported lower downstream resistance in the internal carotid artery in women with polycystic ovary syndrome. This study was designed to assess vascular reactivity by measuring the response to inhalation of 5% carbon dioxide. We studied 34 young women with polycystic ovary syndrome, 15 with symptomless polycystic ovaries, and 18 controls. PMID- 11030302 TI - Pharyngeal carriage of group C and group G streptococci and acute rheumatic fever in an Aboriginal population. AB - Acute rheumatic fever (ARF) and rheumatic heart disease (RHD) represent an autoimmune response to group A streptococcal pharyngitis. In the Aboriginal population of the Northern Territory of Australia, some of the highest rates of ARF in the world have been reported, although throat carriage rates of group A streptococcus in this population are extremely low and symptomatic group A streptococcal pharyngitis is uncommon; by contrast, carriage rates of group C and G streptococci are high. Therefore, we questioned the involvement of these groups in ARF and showed that they have the potential to elicit an autoimmune response that may trigger ARF. PMID- 11030303 TI - Fetal learning and memory. AB - We used fetal habituation to repeated vibroacoustic stimulation to assess fetal memory. After the initial stimulus, we assessed the fetuses 10 min later and again after 24 h. 16 of 19 fetuses habituated rapidly to the stimuli at 10 min (p=0.004) and 24 h (p=0.042) after the initial test. We conclude, therefore, that fetuses are able to learn: they have a short-term memory of at least 10 min, and a long-term memory of at least 24 h. PMID- 11030305 TI - Olympic committee bans doctor after doping case. PMID- 11030304 TI - Prenatal DNA diagnosis of a single-gene disorder from maternal plasma. AB - Achondroplasia is a short-limb disorder caused by a point mutation in a single gene. To diagnose such a disorder prenatally requires the use of invasive procedures such as amniocentesis. However, using PCR and restriction fragment length polymorphism analysis, we were able to detect the mutation in the plasma of a woman carrying a fetus suspected of having achondroplasia. The detection of a fetus-derived mutant gene from maternal plasma may therefore permit non invasive prenatal diagnosis of single-gene disorders. PMID- 11030306 TI - Hunting and logging linked to emerging infectious diseases. PMID- 11030307 TI - Maggot therapy: an alternative for wound infection. PMID- 11030308 TI - Nargile smoking keeps Arabs in Wonderland. PMID- 11030309 TI - European holiday resorts at risk for Legionella. PMID- 11030310 TI - Public-health preparedness for biological terrorism in the USA. PMID- 11030311 TI - Diabete des femmes a barbe: a classic paper reread. PMID- 11030312 TI - Clinical picture. Cutaneous hyalohyphomycosis. PMID- 11030313 TI - Health and the UK Human Rights Act 1998. PMID- 11030314 TI - The DDT question. PMID- 11030315 TI - The DDT question. PMID- 11030316 TI - The DDT question. PMID- 11030317 TI - The DDT question. PMID- 11030318 TI - The DDT question. PMID- 11030319 TI - The DDT question. PMID- 11030320 TI - T-helper imbalance in allergen sensitisation. PMID- 11030321 TI - Difficulty in development of new anxiolytics in Japan. PMID- 11030322 TI - Management of unstable angina and myocardial infarction. PMID- 11030323 TI - False positivity of prenatal Down's syndrome and neural-tube tests in SLE. PMID- 11030324 TI - Effects of statins. PMID- 11030325 TI - Empowerment of individuals to reduce cardiovascular disease. PMID- 11030326 TI - Medical migration. PMID- 11030327 TI - Medical migration. PMID- 11030328 TI - Medical migration. PMID- 11030329 TI - Medical migration. PMID- 11030330 TI - Medical migration. PMID- 11030331 TI - Molecular basis for feedback regulation of bile acid synthesis by nuclear receptors. AB - The catabolism of cholesterol into bile acids is regulated by oxysterols and bile acids, which induce or repress transcription of the pathway's rate-limiting enzyme cholesterol 7alpha-hydroxylase (CYP7A1). The nuclear receptor LXRalpha binds oxysterols and mediates feed-forward induction. Here, we show that repression is coordinately regulated by a triumvirate of nuclear receptors, including the bile acid receptor, FXR; the promoter-specific activator, LRH-1; and the promoter-specific repressor, SHP. Feedback repression of CYP7A1 is accomplished by the binding of bile acids to FXR, which leads to transcription of SHP. Elevated SHP protein then inactivates LRH-1 by forming a heterodimeric complex that leads to promoter-specific repression of both CYP7A1 and SHP. These results reveal an elaborate autoregulatory cascade mediated by nuclear receptors for the maintenance of hepatic cholesterol catabolism. PMID- 11030332 TI - A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. AB - Bile acids repress the transcription of cytochrome P450 7A1 (CYP7A1), which catalyzes the rate-limiting step in bile acid biosynthesis. Although bile acids activate the farnesoid X receptor (FXR), the mechanism underlying bile acid mediated repression of CYP7A1 remained unclear. We have used a potent, nonsteroidal FXR ligand to show that FXR induces expression of small heterodimer partner 1 (SHP-1), an atypical member of the nuclear receptor family that lacks a DNA-binding domain. SHP-1 represses expression of CYP7A1 by inhibiting the activity of liver receptor homolog 1 (LRH-1), an orphan nuclear receptor that is known to regulate CYP7A1 expression positively. This bile acid-activated regulatory cascade provides a molecular basis for the coordinate suppression of CYP7A1 and other genes involved in bile acid biosynthesis. PMID- 11030333 TI - TAC, a TBP-sans-TAFs complex containing the unprocessed TFIIAalphabeta precursor and the TFIIAgamma subunit. AB - Transcription of TATA box-containing genes by RNA polymerase II is mediated by TBP-containing and TBP-free multisubunit complexes consisting of common and unique components. We have identified a highly stable TBP-TFIIA-containing complex, TAC, which is detectable in embryonal carcinoma (EC) cells but not in differentiated cells. TAC contains the TFIIAgamma subunit and the unprocessed form of TFIIAalphabeta, although the processed TFIIAalpha and TFIIAbeta subunits are present in EC cells. TAC mediates transcriptional activation by RNA polymerase II in vivo, even though it does not contain classical TAFs. Formaldehyde cross-linking revealed that in EC but not in differentiated cells, association of TBP with chromatin is strongly enhanced when complexed with TFIIA in vivo. Remarkably, the TFIIAalphabeta precursor is preferentially, if not exclusively, associated with chromatin as compared to the processed subunits present in "free" TFIIA in EC cells. PMID- 11030334 TI - Concerted dephosphorylation of the transcription factor NFAT1 induces a conformational switch that regulates transcriptional activity. AB - NFAT transcription factors are highly phosphorylated proteins that are regulated by the calcium-dependent phosphatase calcineurin. We show by mass spectrometry that NFAT1 is phosphorylated on fourteen conserved phosphoserine residues in its regulatory domain, thirteen of which are dephosphorylated upon stimulation. Dephosphorylation of all thirteen residues is required to mask a nuclear export signal (NES), cause full exposure of a nuclear localization signal (NLS), and promote transcriptional activity. An inducible phosphorylation site in the transactivation domain contributes to transcriptional activity. Our data suggest that dephosphorylation promotes NFAT1 activation by increasing the probability of an active conformation, in a manner analogous to that by which depolarization increases the open probability of voltage-gated ion channels. This conformational switch paradigm may explain modification-induced functional changes in other heavily phosphorylated proteins. PMID- 11030335 TI - Activator-dependent transcription from chromatin in vitro involving targeted histone acetylation by p300. AB - The transcriptional coactivator p300 shows physical and functional interactions with a diverse group of activators and contains an intrinsic acetyltransferase activity whose exact coactivator functions in the acetylation of nucleosomal histones versus other factors are poorly documented. Here, we show that p300 mediates acetyl-CoA-dependent transcription by GAL4-VP16 from a nucleosomal array template, that this involves p300 targeting by GAL4-VP16 and promoter-proximal histone acetylation prior to transcription, and that the affinities of different activators for p300 roughly correlate with corresponding levels of p300-dependent transcription. These results indicate that activators recruit p300 to nucleosomal templates by direct interactions and that bound p300 stimulates transcription, at least in part, by localized histone acetylation. PMID- 11030336 TI - Superhelicity-driven homologous DNA pairing by yeast recombination factors Rad51 and Rad54. AB - Yeast Rad51 recombinase has only minimal ability to form D loop. Addition of Rad54 renders D loop formation by Rad51 efficient, even when topologically relaxed DNA is used as substrate. Treatment of the nucleoprotein complex of Rad54 and relaxed DNA with topoisomerases reveals dynamic DNA remodeling to generate unconstrained negative and positive supercoils. DNA remodeling requires ATP hydrolysis by Rad54 and is stimulated by Rad51-DNA nucleoprotein complex. A marked sensitivity of DNA undergoing remodeling to P1 nuclease indicates that the negative supercoils produced lead to transient DNA strand separation. Thus, a specific interaction of Rad54 with the Rad51-ssDNA complex enhances the ability of the former to remodel DNA and allows the latter to harvest the negative supercoils generated for DNA joint formation. PMID- 11030337 TI - Tn7 transposes proximal to DNA double-strand breaks and into regions where chromosomal DNA replication terminates. AB - We report that the bacterial transposon Tn7 can preferentially transpose into regions where chromosomal DNA replication terminates. DNA double-strand breaks are associated with the termination of chromosomal replication; therefore, we directly tested the effect of DNA breaks on Tn7 transposition. When DNA double strand breaks are induced at specific sites in the chromosome, Tn7 transposition is stimulated and insertions are directed proximal to the induced break. The targeting preference for the terminus of replication and DNA double-strand breaks is dependent on the Tn7-encoded protein TnsE. The results presented in this study could also explain the previous observation that Tn7 is attracted to events associated with conjugal DNA replication during plasmid DNA transfer. PMID- 11030338 TI - Rad54 protein is targeted to pairing loci by the Rad51 nucleoprotein filament. AB - Rad51 and Rad54 proteins are important for the repair of double-stranded DNA (dsDNA) breaks by homologous recombination in eukaryotes. Rad51 assembles on single-stranded DNA (ssDNA) to form a helical nucleoprotein filament that performs homologous pairing with dsDNA; Rad54 stimulates this pairing substantially. Here, we demonstrate that Rad54 acts in concert with the mature Rad51-ssDNA filament. Enhancement of DNA pairing by Rad54 is greatest at an equimolar ratio relative to Rad51 within the filament. Reciprocally, the Rad51 ssDNA filament enhances both the dsDNA-dependent ATPase and the dsDNA unwinding activities of Rad54. We conclude that Rad54 participates in the DNA homology search as a component of the Rad51-nucleoprotein filament and that the filament delivers Rad54 to the dsDNA pairing locus, thereby linking the unwinding of potential target DNA with the homology search process. PMID- 11030339 TI - Checkpoint-dependent activation of mutagenic repair in Saccharomyces cerevisiae pol3-01 mutants. AB - The Saccharomyces cerevisiae DNA polymerase delta proofreading exonuclease defective mutation pol3-01 is known to cause high rates of accumulating mutations. The pol3-01 mutant was found to have abnormal cell cycle progression due to activation of the S phase checkpoint. Inactivation of the S phase checkpoint suppressed both the pol3-01 cell cycle progression defect and mutator phenotype, indicating that the pol3-01 mutator phenotype was dependent on the S phase damage checkpoint pathway. Epistasis analysis suggested that a portion of the pol3-01 mutator phenotype involves members of the RAD6 epistasis group that function in both error-free and error-prone repair. These results indicate that activation of a checkpoint in response to certain types of replicative defects can result in the accumulation of mutations. PMID- 11030340 TI - A global profile of germline gene expression in C. elegans. AB - We used DNA microarrays to profile gene expression patterns in the C. elegans germline and identified 1416 germline-enriched transcripts that define three groups. The sperm-enriched group contains an unusually large number of protein kinases and phosphatases. The oocyte-enriched group includes potentially new components of embryonic signaling pathways. The germline-intrinsic group, defined as genes expressed similarly in germlines making only sperm or only oocytes, contains a family of piwi-related genes that may be important for stem cell proliferation. Finally, examination of the chromosomal location of germline transcripts revealed that sperm-enriched and germline-intrinsic genes are nearly absent from the X chromosome, but oocyte-enriched genes are not. PMID- 11030341 TI - Components of the SWI/SNF complex are required for asymmetric cell division in C. elegans. AB - Asymmetric cell division is a fundamental process that produces cellular diversity during development. We have identified two mutants in C. elegans (psa-1 and psa-4) in which the asymmetry of T cell division is disrupted. psa-1 and psa 4 encode homologs of yeast SWI3 and SWI2/SNF2, respectively, which are components of the SWI/SNF complex. We show by RNA interference assay that homologs of other components of SWI/SNF are also involved in T cell division. psa-1 and psa-4 are likely to be required in the T cell during mitosis to cause asymmetric cell division. Because the SWI/SNF complex is required for asymmetric division in S. cerevisiae, these results demonstrate that at least some aspects of the mechanism of asymmetric cell division are conserved between yeast and a multicellular organism. PMID- 11030342 TI - Requirements for presenilin-dependent cleavage of notch and other transmembrane proteins. AB - Ligand binding to receptors of the LIN-12/Notch family causes at least two proteolytic cleavages: one between the extracellular and transmembrane domains, and the other within the transmembrane domain. The transmembrane cleavage depends on Presenilin, a protein also required for transmembrane cleavage of beta-APP. Here, we have assayed the substrate requirements for Presenilin-dependent processing of Notch and other type I transmembrane proteins in vivo. We find that the Presenilin-dependent cleavage does not depend critically on the recognition of particular sequences in these proteins but rather on the size of the extracellular domain: the smaller the size, the greater the efficiency of cleavage. Hence, Notch, beta-APP, and perhaps other proteins may be targeted for Presenilin-mediated transmembrane cleavage by upstream processing events that sever the extracellular domain from the rest of the protein. PMID- 11030343 TI - Structure and function of Cdc6/Cdc18: implications for origin recognition and checkpoint control. AB - Cdc6/Cdc18 is a conserved and essential component of prereplication complexes. The 2.0 A crystal structure of an archaeal Cdc6 ortholog, in conjunction with a mutational analysis of the homologous Cdc18 protein from Schizosaccharomyces pombe, reveals novel aspects of Cdc6/Cdc18 function. Two domains of Cdc6 form an AAA+-type nucleotide binding fold that is observed bound to Mg.ADP. A third domain adopts a winged-helix fold similar to known DNA binding modules. Sequence comparisons show that the winged-helix domain is conserved in Orc1, and mutagenesis data demonstrate that this region of Cdc6/Cdc18 is required for function in vivo. Additional mutational analyses suggest that nucleotide binding and/or hydrolysis by Cdc6/Cdc18 is required not only for progression through S phase, but also for maintenance of checkpoint control during S phase. PMID- 11030344 TI - Reconstitution of an ATM-dependent checkpoint that inhibits chromosomal DNA replication following DNA damage. AB - Cell cycle checkpoints lead to the inhibition of cell cycle progression following DNA damage. A cell-free system derived from Xenopus eggs has been established that reconstitutes the checkpoint pathway inhibiting DNA replication initiation. DNA containing double-strand breaks inhibits replication initiation in a dose dependent manner. Upon checkpoint activation, a prereplicative complex is assembled that contains ORC, Cdc6, Cdc7, and MCM proteins but lacks Cdc45. The checkpoint is ATM dependent. Cdk2/CyclinE acts downstream of ATM and is downregulated by Cdk2 phosphorylation on tyrosine 15. Cdk2AF/CyclinE is refractory to checkpoint signaling, and Cdc25A overrides the checkpoint and restores DNA replication. This report provides the description of a DNA damage checkpoint pathway that prevents the onset of S phase independently of the transcriptional function of p53 in a vertebrate organism. PMID- 11030345 TI - Class C Vps protein complex regulates vacuolar SNARE pairing and is required for vesicle docking/fusion. AB - In yeast, the Class C Vps protein complex (C-Vps complex), composed of Vps11, Vps16, Vps18, and Vps33, functions in Golgi-to-vacuole protein transport. In this study, we characterized and purified this complex and identified its interaction with the syntaxin homolog Vam3. Vam3 pairs with the SNAP-25 homolog Vam7 and VAMP homolog Vti1 to form SNARE complexes during vesicle docking/fusion with the vacuole. The C-Vps complex does not bind to Vam3-Vti1-Vam7 paired SNARE complexes but instead binds to unpaired Vam3. Antibodies to a component of this complex inhibited in vitro vacuole-to-vacuole fusion. Furthermore, temperature conditional mutations in the Class C VPS genes destabilized Vam3-Vti1-Vam7 pairing. Therefore, we propose that the C-Vps complex associates with unpaired (activated) Vam3 to mediate the assembly of trans-SNARE complexes during both vesicle docking/fusion and vacuole-to-vacuole fusion. PMID- 11030346 TI - Pre-mRNA splicing imprints mRNA in the nucleus with a novel RNA-binding protein that persists in the cytoplasm. AB - We describe a novel RNA binding protein, Y14, a predominantly nuclear nucleocytoplasmic shuttling protein. Interestingly, Y14 associates preferentially with mRNAs produced by splicing but not with pre-mRNAs, introns, or mRNAs produced from intronless cDNAs. Y14 associates with both nuclear mRNAs and newly exported cytoplasmic mRNAs. Splicing of a single intron is sufficient for Y14 association. Y14-containing nuclear complexes are different from general hnRNP complexes. They contain hnRNP proteins and several unique proteins including the mRNA export factor TAP. Thus, Y14 defines novel intermediates in the pathway of gene expression, postsplicing nuclear preexport mRNPs, and newly exported cytoplasmic mRNPs, whose composition is established by splicing. These findings suggest that pre-mRNA splicing imprints mRNA with a unique set of proteins that persists in the cytoplasm and thereby communicates the history of the transcript. PMID- 11030347 TI - In the absence of extrinsic signals, nutrient utilization by lymphocytes is insufficient to maintain either cell size or viability. AB - Without receptor stimulation, cells from multicellular organisms die by apoptosis. Here we show that lymphocytes deprived of receptor stimulation undergo progressive atrophy before commitment to apoptosis. Following loss of receptor engagement, lymphocytes rapidly downregulated the glucose transporter, glut1. This was accompanied by reduction in mitochondrial potential and cellular ATP, suggesting that atrophy resulted from depletion of glucose-derived metabolic substrates. Expression of the antiapoptotic protein, Bcl-X(L), prevented death but not atrophy following either growth factor or glucose withdrawal. In Bcl-X(L) transgenic animals, size and metabolic activity of naive T cells were regulated through the TCR and correlated with TCR-dependent glut1 expression. These data suggest that ligands for cell-specific receptors promote cell survival by regulating nutrient uptake and utilization. PMID- 11030348 TI - Stat5 is essential for the myelo- and lymphoproliferative disease induced by TEL/JAK2. AB - STAT5 is activated in a broad spectrum of human hematologic malignancies. We addressed whether STAT5 activation is necessary for the myelo- and lymphoproliferative disease induced by TEL/JAK2 using a genetic approach. Whereas mice transplanted with bone marrow transduced with retrovirus expressing TEL/JAK2 develop a rapidly fatal myelo- and lymphoproliferative syndrome, reconstitution with bone marrow derived from Stat5ab-deficient mice expressing TEL/JAK2 did not induce disease. Disease induction in the Stat5a/b-deficient background was rescued with a bicistronic retrovirus encoding TEL/JAK2 and Stat5a. Furthermore, myeloproliferative disease was induced by reconstitution with bone marrow cells expressing a constitutively active mutant, Stat5a, or a single Stat5a target, murine oncostatin M (mOSM). These data define a critical role for Stat5a/b and mOSM in the pathogenesis of TEL/JAK2 disease. PMID- 11030349 TI - The TBP-like factor CeTLF is required to activate RNA polymerase II transcription during C. elegans embryogenesis. AB - Metazoans possess two TATA-binding protein homologs, the general transcription factor TBP and a related factor called TLF. Four models have been proposed for the role of TLF in RNA polymerase II (Pol II) transcription: (1) TLF and TBP function redundantly, (2) TLF antagonizes TBP, (3) TLF is a tissue-specific TBP, or (4) TLF and TBP have distinct activities. Here we report that CeTLF is required to express a subset of Pol II genes and associates with at least one of these genes in vivo. CeTLF is also necessary to establish bulk transcription during early embryogenesis. Since CeTLF and CeTBP are expressed at comparable levels in the same cells, these findings suggest CeTLF performs a unique function in activating Pol II transcription distinct from that of CeTBP. PMID- 11030350 TI - TBP-like factor is required for embryonic RNA polymerase II transcription in C. elegans. AB - Recently, a novel family of TATA binding protein (TBP)-like factors (TLFs) have been described in metazoan organisms; however, their function has not yet been elucidated. Using Caenorhabditis elegans (Ce) as a model, we demonstrate that CeTLF is required in vivo for zygotic transcription during embryogenesis. Elimination of CeTLF expression by RNA interference caused embryonic lethality either due to the lack of expression of early patterning genes or to their ectopic expression. Moreover, the absence of CeTLF in vivo prevented the correct soma-specific phosphorylation of RNA polymerase II (Pol II). Thus, CeTLF may positively or negatively regulate Pol II transcription, depending on the developmental stage of the embryo. PMID- 11030351 TI - Transcriptional control: rheostat converted to on/off switch. AB - Individual cells translate concentration gradients of extracellular factors into all-or-none threshold responses leading to discrete patterns of gene expression. Signaling cascades account for some but not all such threshold responses, suggesting the existence of additional mechanisms. Here we show that all-or-none responses can be generated at a transcriptional level. A graded rheostat mechanism obtained when either transactivators or transrepressors are present is converted to an on/off switch when these factors compete for the same DNA regulatory element. Hill coefficients of dose-response curves confirm that the synergistic responses generated by each factor alone are additive, obviating the need for feedback loops. We postulate that regulatory networks of competing transcription factors prevalent in cells and organisms are crucial for establishing true molecular on/off switches. PMID- 11030352 TI - E2F4 and E2F5 play an essential role in pocket protein-mediated G1 control. AB - E2F transcription factors are major regulators of cell proliferation. The diversity of the E2F family suggests that individual members perform distinct functions in cell cycle control. E2F4 and E2F5 constitute a defined subset of the family. Until now, there has been little understanding of their individual biochemical and biological functions. Here, we report that simultaneous inactivation of E2F4 and E2F5 in mice results in neonatal lethality, suggesting that they perform overlapping functions during mouse development. Embryonic fibroblasts isolated from these mice proliferated normally and reentered from Go with normal kinetics compared to wild-type cells. However, they failed to arrest in G1 in response to p16INK4a. Thus, E2F4 and E2F5 are dispensable for cell cycle progression but necessary for pocket protein-mediated G1 arrest of cycling cells. PMID- 11030353 TI - Requirements for cell cycle arrest by p16INK4a. AB - Analysis of tumor-derived mutations has led to the suggestion that p16INK4a, cyclin D1, cdk4, and the retinoblastoma protein (pRB) are components of a regulatory pathway that is inactivated in most tumor cells. Cell cycle arrest induced by p16INK4a, an inhibitor of cyclin D-dependent kinases, requires pRB, and it has been proposed that this G1 arrest is mediated by pRB-E2F repressor complexes. By comparing the properties of primary mouse embryonic fibroblasts specifically lacking pRB-family members, we find that pRB is insufficient for a p16INK4a-induced arrest. In addition to pRB, a second function provided by either p107 or p130, two pRB-related proteins, is required for p16INK4a to block DNA synthesis. We infer that p16INK4a-induced arrest is not mediated exclusively by pRB, but depends on the nonredundant functions of at least two pRB-family members. PMID- 11030354 TI - Crystal structure of a ternary FGF-FGFR-heparin complex reveals a dual role for heparin in FGFR binding and dimerization. AB - The crystal structure of a dimeric 2:2:2 FGF:FGFR:heparin ternary complex at 3 A resolution has been determined. Within each 1:1 FGF:FGFR complex, heparin makes numerous contacts with both FGF and FGFR, thereby augmenting FGF-FGFR binding. Heparin also interacts with FGFR in the adjoining 1:1 FGF:FGFR complex to promote FGFR dimerization. The 6-O-sulfate group of heparin plays a pivotal role in mediating both interactions. The unexpected stoichiometry of heparin binding in the structure led us to propose a revised model for FGFR dimerization. Biochemical data in support of this model are also presented. This model provides a structural basis for FGFR activation by small molecule heparin analogs and may facilitate the design of heparin mimetics capable of modulating FGF signaling. PMID- 11030356 TI - Canine ehrlichioses: an update. AB - The development of molecular biology techniques and methods for the isolation and growth of ehrlichias in tissue culture have greatly facilitated the study of these organisms. The available knowledge on ehrlichias is thus rapidly increasing and in this review recent findings on the epidemiology, transmission, clinical and laboratory signs of infection, diagnosis and treatment of canine ehrlichioses are described. PMID- 11030355 TI - Harnessing the ubiquitination machinery to target the degradation of specific cellular proteins. AB - The functional characterization of a specific gene, or its protein product, often relies on assessing the consequences of its elimination, usually accomplished by gene knockout, ribozyme, antisense, or RNA-mediated interference (RNAi) technologies. The selective degradation of cellular proteins is mediated primarily by the ubiquitin-proteasome pathway. Manipulation of the ubiquitin dependent proteolytic machinery to eliminate specific gene products at the protein level has been previously attempted with some success in vitro; however, the in vivo efficacy of this approach has not yet been achieved. Here we report successful engineering of the substrate receptor of a major ubiquitin-proteolytic machinery to direct the degradation of otherwise stable cellular proteins both in yeast and in mammalian cells. PMID- 11030357 TI - Use of herbal remedies by small-scale farmers to treat livestock diseases in Central Eastern Cape Province, South Africa. AB - The use of herbal remedies by African farmers in the treatment of livestock diseases was investigated using a range of methods, including group interviews, structured interviews, semi-structured interviews and participant observation. The study was concerned mainly with the 'why' and 'how' of herbal remedy use among African livestock holders farming on communally-owned land in central Eastern Cape Province. We found that the use of herbal remedies was still widespread because these remedies were cheap, locally available and convenient to administer. Farmers also used herbal remedies because they did not have knowledge of conventional remedies or found these too expensive and therefore used herbal remedies as a last resort. Most of the farmers interviewed (73%) had used some form of herbal remedy. A small number of farmers did not use herbal remedies at all. They rejected them because they had bad experiences in the past or because they considered the practice outdated and socially inferior. Often farmers used herbal remedies in combination with conventional ones, but a substantial number relied on herbal remedies only. Farmers obtained herbal remedies principally by preparing their own or by purchasing from herbalists. Different plant parts were used to prepare herbal remedies in the form of infusions, decoctions, powders, pastes, and juices from fresh plant material. Preparations were made from a single plant or from a range of plants. Application of a remedy was by different routes and methods, depending on the perceived cause of the disease condition. Topical applications were used for skin conditions, powders were rubbed into incisions, drenches were common in the treatment of systemic conditions, and drops to treat ears and eyes. We identified several aspects of herbal remedy utilisation that could be subject to improvement, which included the absence of standard dose regimes, the potential toxicity of certain compounds present in plants used, over-exploitation of the plant resource and a lack of interest by the youth to learn the art. PMID- 11030358 TI - Animal husbandry in Moretele 1 of North-West Province: implications for veterinary training and research. AB - Little is known regarding the keeping of animals in the Moretele 1 area of North West Province, South Africa. Therefore, the status and dynamics of animal husbandry, as well as a general assessment of the needs of animal owners in this area were researched. Results of the investigation will be used to make recommendations for improved veterinary extension servicing in the area. Semi structured interviews, based on discussions with relevant stakeholders in the community and a resultant problem conceptualisation, were undertaken at 266 randomly selected households in 51 villages and centres in the area, after which the data was checked and verified before being captured and analysed. The findings reveal that within the field of veterinary extension delivery: 1) there is a demand for visual and written extension material, 2) the extension services must function where clients reside, 3) limitations in terms of infrastructure are present and should be addressed through partnerships and coordination amongst all the role-players in the Moretele 1 area, and 4) cattle and poultry are the most important of the animal species and should be the focus points of extension, but the need to curb zoonotic disease should not be disregarded. In this regard veterinary clinics, private veterinarians and other role-players should be used in partnership with extension workers. Lastly, the veterinary clinic is regarded as helpful in many respects by the community consulted and the service should be upgraded and made available to a wider client base, especially where private and state veterinarians are unavailable or too expensive in such resource-limited communities. PMID- 11030359 TI - Ultrastructural characteristics of ostrich eggshell: outer shell membrane and the calcified layers. AB - The ultrastructure of the eggshell of the domestic hen has been well researched and structural studies of other avian species, such as the ostrich, often base their interpretation of egg shell structure on that of the chicken. In the ostrich, lowered hatchability and hatching trauma may be due to shell ultrastructural abnormalities. In the present study the ultrastructure of the calcified portion, and the outer shell membrane (OSM), of domesticated ostrich eggshells was investigated using standard electron microscopic techniques. Transmission and scanning electron microscopy studies demonstrated intimate contact between cup-shaped structures present on the OSM and the mammillary layer of the calcified portion of the shell. The initial calcium carbonate growth of the calcified shell was of a dendritic nature with nucleation sites on the surface of the cup's contents. The dendritic growth gave way to a more randomly orientated, smaller crystallite growth structure, which changed in form as it neared the vertical crystal layer (VCL). The VCL is described as being both amorphous and 'crumbly' depending on the plane of fracture. These observations suggest that firstly, initial calcification is contained within the cups and is then directed outwards to form the shell and that secondly, the VCL may contain an evolutionary, calcified cuticular layer. These observations serve as a baseline for studies investigating the effect of shell structure and strength on hatchling trauma and the influence of maternal diet. PMID- 11030360 TI - An appropriate method for extracting the insect repellent citronellol from an indigenous plant (Pelargonium graveolens L'Her) for potential use by resource limited animal owners. AB - Veterinary needs appraisals in rural, peri-urban and urban areas have indicated a need for affordable and accessible veterinary health care. It was also found that farmers and animal owners used indigenous plants for treating animals. In Africa, insects such as Culex, Culicoides and Stomoxys may transmit diseases, cause irritation to animals or prevent wound healing. Insect repellents used topically are generally safer and cheaper than insecticides. Using readily available commercial sources of ethanol 43% v/v (brandy and cane spirits), it was shown that citronellol could be extracted from uncrushed leaves of the indigenous shrub Pelargonium graveolens L'Her. Efficacy of extraction was compared to that using reagent grade absolute ethanol. The peak concentration of citronellol was achieved within 7 days of extraction and thereafter remained constant for 4 months. Extraction methods using tap water and cooking oil were not successful. The extraction was also less successful when the leaves were crushed or macerated before being placed into ethanol. Gas chromatography was used to monitor the concentration of citronellol in the different extracts. PMID- 11030361 TI - Coccidian oocyst and nematode egg counts of free-ranging African buffalo (Syncerus caffer) in the Kruger National Park, South Africa. AB - Faecal specimens collected in the Kruger National Park from 103 African buffaloes (Syncerus caffer) up to 1 year old and 283 buffaloes older than 1 year were examined for the presence of coccidian oocysts and nematode eggs. Most specimens from animals older than 1 year had negative coccidian oocyst counts. Positive specimens from younger animals had significantly higher coccidian oocyst counts than those from older animals. No such difference was found for nematode egg counts. PMID- 11030362 TI - Efficacy of a commercially available coryza vaccine against challenge with recent South African NAD-independent isolates of Haemophilus paragallinarum in chickens. AB - In South Africa the incidence of NAD-independent Haemnophilus paragallinarum isolation from clinical cases is increasing. This study was carried out to test whether a commercially available coryza vaccine (Nobilis Coryza, Intervet International BV) could protect chickens against challenge with recent NAD independent isolates. SPF chickens were vaccinated twice at 3 and 7 weeks of age and were challenged at 9 weeks of age with 5 different NAD-independent isolates of serotype A or C-3. The results after challenge show that the coryza vaccine induces good protection against challenge with the different South African NAD independent isolates of H. paragallinarum, including serotype C-3. PMID- 11030363 TI - Evaluation of two PCR-based procedures for typing Clostridium perfringens. AB - Two polymerase chain reaction (PCR)-based procedures for typing Clostridium, perfringens, which affects most domestic animals, were compared and evaluated for efficiency as substitute to the guinea-pig intradermal test routinely used in our laboratory, namely a multiplex PCR and a protocol based on the individual amplification of gene sequences specific for each toxin. Reference isolates of C. perfringens types A, B, C and D as well as cultures from clinical specimens were tested. The sensitivity and specificity of the PCR was confirmed on reference isolates. There was similarity in results on 43 of the 46 samples typed by all 3 methods. Clear results were obtained by PCR on 5 clinical samples that showed either equivocal or weak skin reactions in guinea-pigs. The multiplex PCR protocol, in combination with the evaluation of bacterial growth, is a better alternative to in vivo toxin typing, since C. perfringens can only be incriminated as cause of a disease when it is present in large numbers in the intestine. PMID- 11030364 TI - A retrospective study on some reproductive parameters of German shepherd bitches in Kenya. AB - Data relating to reproductive parameters of German shepherd bitches were collected from registered German shepherd dog (GSD) breeders with information kept over a 15-year period (1982-1997). The information obtained was verified using the East African Kennel Club records. A total of 594 bitches from 280 breeders were recorded. From these, 798 heats were observed, 594 of which were used for breeding, resulting in 3592 puppies. The mean age at puberty was 519.0 +/- 41 days. Heats occurred throughout the year, although significantly (P < 0.05) higher and lower incidences were observed in October and April respectively Pregnancy significantly (P < 0.01) increased interoestrous interval, which was 247.8 +/- 99.6 and 183 +/- 52 days among bred/pregnant and non-bred bitches respectively. Most bitches in oestrus (73.7%) were bred, and breeding was carried out throughout the year, with a distribution closely related to that of heat incidence. Subsequently, whelping occurred throughout the year, and 95.5% of the bitches that were mated whelped. A mean gestation period of 60.6 +/- 5.1 days was observed. The mean litter size was 6.4 +/- 0.4 puppies, and did not differ significantly between months. The preweaning losses were low, with 2.3% stillbirths, 0.9% culls and 11.4% mortalities. PMID- 11030365 TI - Von Willebrand's disease in the German shepherd dog. AB - Two litters of German shepherd dogs were evaluated for a haemorrhagic tendency that was characterised by excessive bleeding from the umbilicus at birth, haemorrhage and haematoma formation at vaccination, excessive bruising, and lameness due to haemarthrosis. Platelet counts, clotting times and Von Willebrand's factor (VWF) assays were assessed in all dogs. Factor VIII determination was performed in 1 puppy and its parents. Based on the clotting times and VWF assay, 6 puppies (4 male and 2 female) showed type I Von Willebrand's disease (VWD), 5 (4 male and 1 female) possible type II VWD, and 4 were unaffected. One puppy with possible type II VWD had very low factor VIII activity; its sire had a normal factor activity, whereas the dam was in the low normal range. This article reports type I and possible type II VWD in 2 related litters of German shepherd dogs, the latter being rare in German shepherd dogs. PMID- 11030366 TI - Progressive atrophic rhinitis in a medium-scale pig farm in Kiambu, Kenya. AB - Forty-two pigs in a herd of 117 displayed various clinical signs of progressive atrophic rhinitis. The main signs included sneezing, coughing, lachrymation, serous to mucopurulent nasal discharge, and nasal bleeding in 1 pig. Three pigs had lateral deviation of the snout, while 4 had brachygnathia superior with obvious deformation of the face. Four acutely affected weaner pigs appeared weak, while the 7 chronically-affected pigs appeared smaller than their apparently unaffected penmates of the same age. Treatment of the acutely affected pigs with long-acting oxytetracycline at 20 mg/kg body weight intramuscularly, repeated once after 7 days, reduced the severity but did not clear the sneezing from all the pigs. Fifteen pigs were slaughtered 2 months after the clinical diagnosis was made. The carcasses of the chronically affected pigs were about 15% lighter than those of the apparently normal pigs of the same age and from the same pen, which translated to a loss of 921.00 Kenya shillings per pig (US$13.7). Diagnosis of progressive atrophic rhinitis was confirmed by sectioning the snouts of randomly selected slaughtered pigs with obvious deformation of the snout. Sections were made at the level of the 1st/2nd upper premolar tooth. Varying degrees of turbinate atrophy, from mild to complete, were noted. Histopathology of the turbinates revealed metaplasia of nasal epithelium and fibrosis in the lamina propria. PMID- 11030367 TI - Arrhythmogenic right ventricular cardiomyopathy in a dog. AB - An 8-month-old Labrador retriever bitch was evaluated for sudden-onset, progressive abdominal distension. Physical examination revealed an exaggerated inspiratory effort, severe ascites, bilateral jugular vein distension, and hypokinetic femoral arterial pulses. Thoracic auscultation detected tachycardia with muffled heart sounds, without audible cardiac murmurs. Thoracic radiographs identified severe right ventricular enlargement and pleural effusion. The electrocardiogram was consistent with incomplete right bundle branch block or right ventricular enlargement. Echocardiography demonstrated severe right ventricular and atrial dilation, secondary tricuspid regurgitation, and thinning and hypocontractility of the right ventricular myocardium. Left heart chamber sizes were slightly decreased, with normal left ventricular contractility. A diagnosis of arrhythmogenic right ventricular cardiomyopathy was reached, based on the characteristic clinical, electrocardiographic, radiographic and echocardiographic findings, and the exclusion of other causes of isolated right ventricular failure. Treatment effected good control of clinical signs, until acutely decompensated congestive right heart failure led to euthanasia after 4 months. Arrhythmogenic right ventricular cardiomyopathy is a well-described clinical entity in humans, and has previously been documented in 3 male dogs. The condition is characterised by progressive fibro-adipose replacement of right ventricular myocardium, while the left ventricle usually remains unaffected. It should be considered a differential diagnosis in any young dog presented with isolated right heart failure, syncope, or unexplained ventricular tachyarrhythmias. This article reports the 1st case of arrhythmogenic right ventricular cardiomyopathy in a female dog, and highlights its echocardiographic features. PMID- 11030368 TI - Allergy and neonatal care. PMID- 11030369 TI - Does sensitization through the skin occur? PMID- 11030370 TI - Eosinophil apoptosis--pathophysiologic and therapeutic implications. PMID- 11030371 TI - The initial care of newborn infants and subsequent hay fever. AB - BACKGROUND: Patterns of neonatal exposure to microorganisms have changed substantially over the last 100 years, and it has been suggested that this has influenced the risk of immune-mediated disease. Using a proxy measure, we tested the hypothesis that the initial handling of newborn infants, which is known to affect the pattern of exposure to microorganisms, may alter the risk of developing subsequent atopy, as indicated by hay fever. METHODS: Analysis was performed on 5,519 members of the 1970 British Cohort Study, a nationally representative birth cohort. Cohort members with hay fever were identified at intervals up to the age of 26 years. Details of neonatal care and childhood circumstances were recorded prospectively. Those who had spent their first night away from their mother in the communal infant nursery were selected as likely to have experienced atypical exposure compared with infants who remained with their mother. Adjustment was made for potential confounding factors in infancy and childhood by multiple logistic regression analysis. RESULTS: Unadjusted relative odds (with 95% CI) for developing hay fever among those spending the first night in the communal nursery, when compared with other infants who remained with the mother, were 1.48 (1.23-1.77), P<0.001. Comprehensive adjustment for the potential confounding factors, including feeding practices on the first day of life, markers of social and material circumstances, and region, did not substantially alter this relationship, with adjusted relative odds of 1.31 (1.08 1.59), P=0.005. CONCLUSIONS: While our proxy measure is associated with an increased risk of hay fever, further research is required to confirm that this is due to the pattern of infectious exposure in very early life. The results are consistent with the hypothesis that the first challenges are particularly important in the development of the newborn infant's immune system. PMID- 11030372 TI - Ash (Fraxinus excelsior)-pollen allergy in central Europe: specific role of pollen panallergens and the major allergen of ash pollen, Fra e 1. AB - BACKGROUND: The role of ash (Fraxinus excelsior) pollen as a cause of spring pollinosis in central Europe has received little attention. It is not clear whether ash pollen is a primary cause of sensitization or whether it is implicated through cross-sensitization to other pollens. METHODS: Over a 22-month period, ash pollen was included in a screening series for inhalant allergies. Pollen data were documented from 1976 through 1999. The frequency of IgE-binding to the ash-specific allergen Fra e 1 and pollen panallergens, respectively, was compared by Western blot between mono- (n = 6), oligo- (n = 16), and polysensitized (n=25) patients. RESULTS: Of 5,416 consecutive patients sensitized to any pollen, 920 (17.6%) had a positive skin prick test to ash. Total pollen counts varied extensively between years (229-5,351) as did peak concentrations (23-837 grains/m3/24 h). Western blotting revealed Fra e 1 sensitization in 100% of monosensitized, 93% of oligosensitized, but only 44% of polysensitized patients. IgE against profilins (Fra e 2), Ca-binding proteins (Fra e 3), and carbohydrate epitopes in the three groups was found in 0/0/17%, 0/19/31%, and 32/72/60%, respectively. At least 50% of sera from patients with Fra e 1 sensitization did not bind with the protein in Western blots under reducing conditions. CONCLUSIONS: Ash pollen should be considered a relevant factor and distinct entity in spring pollinosis. In all, only 20% of positive skin tests to ash appear to result from cross-sensitization to pollen panallergens. PMID- 11030373 TI - ICAM-1 and alpha3beta1 expression by bronchial epithelial cells and their in vitro modulation by inflammatory and anti-inflammatory mediators. AB - BACKGROUND: Adhesion molecules are involved in inflammatory and repair processes of the bronchial epithelium. ICAM-1 is mainly involved in inflammatory reactions, whereas integrins, such as alpha3beta1, are mainly involved in repair processes. METHODS: Using bronchial biopsies from 10 asthmatics and eight controls, we first evaluated by immunohistochemistry expression of alpha3beta1 and ICAM-1 in intact and damaged epithelium. Then, using the human pulmonary epithelial cell line WI 26 VA, we studied, by flow-cytometry, the modulation of ICAM-1 and alpha3beta1 expression, and, by ELISA, the release of fibronectin by proinflammatory cytokines, such as IL-5, and anti-inflammatory cytokines, such as IL-4, TGF-beta, and EGF. RESULTS: alpha3beta1 expression was slightly higher in asthma than in controls, as well as in damaged epithelium than in undamaged epithelium. ICAM-1 expression was higher in asthma than in controls, and similarly distributed in intact or damaged epithelium. In vitro, alpha3beta1 was significantly increased by TGF-beta, EGF, and IL-4, and significantly decreased by IL-5. Fibronectin release was significantly increased by TGF-beta and IL-4, unchanged by EGF, and slightly but significantly decreased by IL-5. ICAM-1 expression was significantly decreased by TGF-beta and IL-4, unchanged by EGF, and significantly increased by IL-5. CONCLUSIONS: These differences in adhesion molecule expression and fibronectin release may be important in epithelial cell inflammation and repair. PMID- 11030374 TI - What safety measures need to be taken in oral food challenges in children? AB - BACKGROUND: Food allergens are often accused of causing numerous ailments. This is particularly true for the pediatric population, where the incidence of food allergy is four times as high as in adults. As food challenges may provoke life threatening reactions, intensive safety measures need to be taken during provocation, and prompt medical intervention may become necessary. METHODS: We retrospectively evaluated 349 oral challenges in 204 children with atopic dermatitis, looking for criteria to help the physician decide which patients need medical intervention. RESULTS: A total of 178 (51%) oral food challenges with the four allergens (cow's milk [CM], hen's egg [HE], wheat, and soy) showed a positive clinical reaction. Of these, 120 (67%) needed medical intervention. In 42 (35%) cases, intervention was parenteral, and oral medication was given in 78 (65%) cases. There was a strong positive correlation (90%) between the level of specific IgE and the need for medical intervention (> or = 17.50 kU/l for CM, wheat, and soy; > or = 3.50 kU/l for HE). Patient history of food allergy was an indicator of the need for medical intervention (P = 0.01). A positive patient history and a high level of specific IgE were significantly (P=0.003) associated with parenteral medication in HE. CONCLUSIONS: Patient history of food allergy is a reliable indicator of the need for medical intervention in the cases of CM, wheat, and soy regardless of the level of specific IgE. With HE, a positive patient history plus a high level of specific IgE significantly indicates the need for parenteral medication. On the basis of our results, we recommend establishing intravenous access in children with a level of specific IgE of > or = 17.50 kU/l (CAP class 4) to CM and wheat, or with specific IgE of > or =3.50 kU/l (CAP class 3) to HE. PMID- 11030375 TI - Symptoms suggestive of atopic rhinitis in children aged 6-9 years and the indoor environment. AB - BACKGROUND: We aimed to investigate the influence of indoor factors on the prevalence of symptoms suggestive of atopic rhinitis in children aged 6-9 years in Upper Austria. METHODS: We analyzed the results from an extended ISAAC (International Study of Asthma and Allergies in Childhood) questionnaire, answered by the parents, about indoor environment and symptoms strongly suggesting atopic rhinitis. This was defined as having reported a running, obstructed, or itchy nose apart from having a cold in the last year. The overall response rate was 93.4%. After excluding 6,016 children (17.1%) with changed indoor environment (due to allergies in the family), we analyzed the remaining subsample of 18,606 questionnaires. RESULTS: The following factors were associated with an increased risk: mother's smoking during pregnancy and/or during time of breast-feeding (OR 1.28; CI 1.07-1.52), synthetic bedding (OR 1.21; CI 1.09-1.36), dampness/mold at home (OR 1.51; CI 1.31-1.74), central heating with gas (OR 1.75; CI 1.06-2.87), and space heating (OR 1.66; CI 1.01 2.98). Cooking with wood (OR 0.62; CI 0.46-0.84) was negatively associated with symptoms. CONCLUSIONS: The indoor environment plays a role in the symptoms of atopic rhinitis in children. However, the population-attributable risks were not particularly high; they were between -2.7% and 9% for the various exposures considered in this study. PMID- 11030376 TI - Anti-FcepsilonRIalpha serum autoantibodies in different subtypes of urticaria. AB - BACKGROUND: In recent years, a histamine-releasing anti-FcepsilonRIalpha autoantibody has been demonstrated in about one-third of patients with chronic urticaria. However, its clinical significance is still unclear. The objective was to detect a possible correlation between the occurrence of the anti FcepsilonRIalpha autoantibody and the clinical type or cause of urticaria. METHODS: Sera from 66 consecutively seen in- and outpatients with various types of urticaria and five healthy controls were examined for the presence of anti FcepsilonRIalpha autoantibodies with a sandwich ELISA technique. In addition, basophil histamine release was studied in 13 autoantibody-positive sera. RESULTS: Anti-FcepsilonRIalpha autoantibodies were found in 17/48 patients with chronic urticaria, in 2/4 with angioedema, in 1/2 with urticarial vasculitis, and in 2/11 with dermographic urticaria. However, no anti-FcepsilonRIalpha autoantibodies were detected in acute, cold, or delayed-pressure urticaria; in urticaria pigmentosa; or in normal controls. Of all chronic urticaria patients, 22 were classified as idiopathic since no underlying cause could be found. Of this group, seven were seropositive for anti-FcepsilonRIalpha. However, anti-FcepsilonRIalpha was also found in patients who went into remission after treatment of identified causes; namely, in one with type I allergy, one with drug intolerance, one with Helicobacter infection, and six with food intolerance. The autoantibody was also detected in 2/4 patients with associated autoimmune diseases. Functional activity was shown in basophil histamine release in 3/4 autoantibody-positive sera of patients with chronic idiopathic urticaria and in 4/6 autoantibody-positive sera of patients who went into remission after the treatment of underlying causes. CONCLUSIONS: These data confirm that anti-FcepsilonRIalpha autoantibodies in urticaria are mostly found in chronic urticaria. Furthermore, their detection independently of the apparent cause of the urticaria suggests that as yet unidentified mechanisms must be operative, possibly related to the chronic inflammatory process and/or individual predispositions that favor their induction. PMID- 11030377 TI - IgE binding to almond proteins in two CAP-FEIA-negative patients with allergic symptoms to almond as compared to three CAP-FEIA-false-positive subjects. AB - BACKGROUND: Allergy to almonds has been frequently reported, but data on the identification of the almond allergens, as well as on the reliability of the methods for in vitro detection of specific IgE for these allergens, are scant. This study aimed to identify the almond allergens and to evaluate the reliability of the CAP-FEIA as the standard system for detection of almond-specific IgE with clinical significance. METHODS: Immunoblotting performed with an almond-protein extract was carried out on the sera of five patients who had previously been tested by the CAP-FEIA system; two of these patients had tested negative with the CAP-FEIA system but suffered life-threatening laryngeal edema after eating almonds, whereas the other three subjects, who had tested positive with CAP-FEIA, did not present any symptoms subsequent to almond ingestion. RESULTS: The sera of the two symptomatic CAP-FEIA-negative patients had IgE that bound only to a 37 kDa protein in immunoblotting. On the contrary, the sera of the three asymptomatic subjects all showed IgE binding to two almond proteins of 62 and 50 kDa, corresponding to the glycosylated components of the extract. CONCLUSIONS: The results here presented suggest that, at least for the examined subjects, the positivity to almond, as measured with a standard laboratory method, is due to the presence of the 62/50-kDa glycoproteins with little or no immunologic significance, and not to the binding to the 37-kDa polypeptide, which appears to be a true almond allergen. PMID- 11030378 TI - Gene polymorphisms of endothelial nitric oxide synthase and angiotensin converting enzyme in patients with asthma. AB - BACKGROUND: Nitric oxide, including that produced by endothelial constitutive nitric oxide synthase (ecNOS), may regulate vascular and airway tone in the lungs and may influence various aspects of airway homeostasis. Angiotensin-converting enzyme (ACE) is expressed at high levels in the lungs and plays a role in the metabolism of angiotensin II, bradykinin, and substance P, all of which are potentially involved in the pathogenesis of asthma. An insertion-deletion polymorphism of the ACE gene has been shown to be associated with enzyme activity levels of ACE. To examine the possible involvement of the ecNOS and/or ACE genes as the genetic basis of bronchial asthma, we investigated whether there was any association between bronchial asthma and polymorphisms of the ecNOS and/or ACE genes. METHODS: A total of 310 patients with bronchial asthma and 121 healthy subjects took part in this study. The ecNOS and ACE genotypes were determined in all subjects by polymerase chain reaction. RESULTS: 1) The distribution of one genotype (bb) of ecNOS was significantly higher in the asthma group than in the control population. 2) The ACE genotype distribution was not significantly different between the control and the asthma groups. 3) In asthmatic patients, the ACE and ecNOS genotype distribution did not differ significantly among groups of patients with different severities of asthma. CONCLUSIONS: These results suggest that polymorphisms of the ecNOS gene, but not the ACE gene, may be associated with the development of asthma. However, the severity of asthma may not be influenced by polymorphisms of the ecNOS and ACE genes. PMID- 11030379 TI - Urinary eosinophil protein X in children with atopic dermatitis: relation to atopy and disease activity. AB - BACKGROUND: Parameters of eosinophil inflammation have been suggested as markers of disease activity in atopic dermatitis (AD), but the value of urinary eosinophil protein X (U-EPX) in children with AD, as well as the influence of allergic sensitization, is not known. METHODS: We measured U-EPX in 59 atopic and 29 nonatopic children with mild (n = 32), moderate (n = 34), and severe (n = 22) AD, as well as in 64 controls. RESULTS: U-EPX was increased in children with AD (110; 67-164 microg/mmol creatinine, median; quartiles) compared to controls (62; 41-95, P<0.001). Children with mild (97; 63-164, P < 0.01), moderate (108; 67 157, P < 0.01), and severe disease (152; 99-202, P < 0.001) had levels of U-EPX higher than controls. U-EPX was significantly higher in children with severe AD than in mild and moderate disease (P < 0.05 for both). Children with AD and a positive skin prick test (120; 81-176) had higher levels of U-EPX than children with a negative skin prick test (87; 56-155, P<0.05). CONCLUSIONS: U-EPX is significantly increased in children with AD and may reflect disease activity. U EPX may also reflect differences in eosinophil activation between those sensitized and those not sensitized to common allergens. PMID- 11030380 TI - Intolerance of osteosynthesis material: evidence of dichromate contact allergy with concomitant oligoclonal T-cell infiltrate and TH1-type cytokine expression in the peri-implantar tissue. AB - BACKGROUND: We report on a 37-year-old man without history of previous allergic disease who developed an aseptic intolerance reaction to a chromium-cobalt alloy, with local discomfort, loosening, and absence of fracture healing. Both in vivo and in vitro allergoimmunologic diagnostic tests were performed. METHODS: Patch testing was done with a European standard series. Specific serum IgE was measured by CAP-FEIA. In addition to immunohistology (APAAP method), peri-implantar tissue was further analyzed by PCR to determine T-cell-receptor-gamma rearrangement and thus the potential clonal (antigen-driven) T-cell repertoire. The actual tissue mRNA expression for IL-4, IL-6, and IFN-gamma was visualized by RT-PCR. RESULTS: Skin testing gave a delayed-type reaction to dichromate. Specific serum IgE to natural rubber latex and grass pollen was found--but without clinical symptoms. Immunohistology revealed a monocytic and dense T-cell infiltrate. The latter, instead of being random, showed an oligoclonal T-cell receptor rearrangement. In addition, there was TH1-type mediator expression (IL-6 and IFN-gamma, but not IL 4). CONCLUSIONS: Skin test, examination of peri-implantar tissue, and the prompt healing after replacement of the osteosynthesis material suggest an allergic reaction. PCR analysis of peri-implantar tissue can further help to identify and understand allergy-mediated implant intolerance reactions. PMID- 11030381 TI - Measles and atopy in Finland. PMID- 11030382 TI - Allergic asthma to shiitake and oyster mushroom. PMID- 11030383 TI - Anaphylaxis to cherimoya. PMID- 11030384 TI - Postoperative rash to ceftriaxone. PMID- 11030385 TI - Predictors of Anisakis simplex symptoms. PMID- 11030386 TI - Sensitization to latex and burns. PMID- 11030387 TI - Rhinitis and nasal continuous positive air pressure? PMID- 11030388 TI - Occupational allergy to papain. PMID- 11030389 TI - Long persistence of IgE antibody to cefaclor. PMID- 11030390 TI - MBP deposition in eosinophilic gastroenteritis. PMID- 11030391 TI - Hypersensitivity to formaldehyde. PMID- 11030392 TI - Minimally invasive orthognathic surgery: endoscopic vertical ramus osteotomy. AB - The endoscopic procedure for placement and activation of a distraction device for mandibular advancement has been previously reported. The purpose of this study was to demonstrate the feasibility of endoscopic exposure, dissection and osteotomy for mandibular set-back. Two cadaver and three anesthetized minipigs were used in this study. Access to the mandibular ramus was achieved through a 2.0 cm submandibular incision. The dissection was carried sharply to the mandible and completed in the subperiosteal plane. Visualization was achieved using a 2.7 mm diameter endoscope (Karl Storz, Germany). Landmarks were identified and a custom-made retractor was inserted into the sigmoid notch. A vertical ramus osteotomy was created (bilaterally) from the sigmoid notch to the mandibular angle. The mandible was set back (average 6 mm) and fixed using 2.0 mm diameter, bicortical screws. Live animals were sacrificed three weeks postoperatively. The mandibles were examined clinically and radiographically to verify proper osteotomy position and clinical union. In all animals, exposure, accurate identification of landmarks, osteotomy placement and screw fixation were achieved. In the live animals (n=3), union between the proximal and distal segments was documented by clinical and radiographic examination. This study demonstrates the feasibility of the EVRO procedure for mandibular set-back in a minipig model. PMID- 11030393 TI - Revascularized muscle transfer for facial reanimation after long-standing facial paralysis. AB - The aim of the present study was to evaluate the results of a group of patients who had received free vascularized muscle transfer to the face for the treatment of long-standing facial paralysis. In 15 patients, neurovascular transfer of 16 segments of the gracilis muscle to the cheek was performed for reanimation of oral commissure movement. Voluntary muscle activation was evaluated by electromyography (EMG) and clinical function was assessed both metrically from frontal photographs and by a rating system with 6 categories. Patient satisfaction was explored through a self-administered questionnaire of general health (SF-36 health survey). The average follow-up period was 49.3 months. All patients showed voluntary activation of the grafted muscles in the EMG. Symmetry of the static and dynamic position of the oral commissure at rest and under function was considerably improved. In general, however, there was a tendency for undercorrection particularly in a vertical direction. Under function, the excursion of the oral commissure on the grafted side reached 63.7% and 65.5% of the nonparetic side in vertical and horizontal directions, respectively. Subjective rating of patient well-being showed that the scores were higher than or comparable to those of healthy adults in six of eight areas of evaluation. PMID- 11030394 TI - Computer-assisted three-dimensional surgical planing and simulation. 3D soft tissue planning and prediction. AB - The purpose of this paper is to report a new technique for three-dimensional facial soft-tissue-change prediction after simulated orthognathic surgical planning. A scheme for soft tissue deformation, "Computer-assisted three dimensional virtual reality soft tissue planning and prediction for orthognathic surgery (CASP)", is presented. The surgical planning was based on three dimensional reconstructed CT visualization. Soft tissue changes were predicted by two newly devised algorithms: Surface Normal-based Model Deformation Algorithm and Ray Projection-based Model Deformation Algorithm. A three-dimensional color facial texture-mapping technique was also used for generating the color photo realistic facial model. As a final result, a predicted and simulated patient's color facial model can be visualized from arbitrary viewing points. PMID- 11030395 TI - Long-term evaluation following temporomandibular joint arthroscopy with lysis and lavage. AB - Although temporomandibular joint arthrocentesis has been utilized for almost 20 years, most papers which evaluate the success of this procedure present relatively short-term data (less than two years). The purpose of this paper is to present the longer-term condition of 22 patients who were re-examined 2 to 10.8 years after arthroscopic surgery with lysis and lavage. Assessment of symptoms reported by the patient as well as of objective signs noted on clinical examination confirms resolution of pain on movement and increased vertical opening. These findings suggest that arthroscopic surgery of the temporomandibular joint is successful in the long term for patients with painful motion. PMID- 11030396 TI - Transcaruncular approach for reconstruction of medial orbital wall fracture. AB - Medial orbital wall fractures can cause horizontal diplopia and enophthalmos. Therefore, reconstruction of displaced medial wall fractures should be considered. We used a transcaruncular approach in five male patients to reconstruct the medial orbital wall after acute injuries and also as a secondary procedure for enophthalmos correction. Four of these patients had a concomitant orbital floor fracture. The incision was made in the caruncule and extended in the conjunctiva superior and inferior into the fornices for 10-12 mm. The tissue was bluntly dissected in an anteroposterior direction. The periosteum was incised dorsal of the posterior lacrimal crest and after elevation of the periosteum, the fractured orbital wall was visible. Transplants up to a height of 2 cm could be inserted for reconstruction of the medial orbital wall. In the cases of acute trauma, the medial wall was reconstructed using a resorbable polydioxanone plate. Cortical bone was used for the reconstruction of late enophthalmos. No postoperative complications were found. The transcaruncular approach gave a rapid entry to the fractured medial orbital wall without a visible scar. PMID- 11030397 TI - Lingual nerve protection during surgical removal of lower third molars. a prospective randomised study. AB - A prospective randomised study was designed to evaluate the efficacy of protecting the lingual nerve by subperiosteal insertion of a retractor in 300 patients. All operations were performed under local anaesthesia and only one molar was removed at a time. In one group, the lower third molar was removed with protection of the lingual nerve and in another group, without protection. Molars which did not need to be sectioned were excluded from the study. The position of the molar, the degree of surgical difficulty and patient gender and age were also recorded. The results indicate 1.33% incidence of temporary lingual nerve dysaesthesia--this being low in comparison with other studies. No permanent disturbances were found. The intergroup percentage difference was not significant. It is suggested that routine application of a lingual protecting instrument during surgical removal of a third molar is not necessary in the hands of an experienced surgeon. PMID- 11030398 TI - Primary thinning of the myocutaneous vastus lateralis flap. AB - To expand the indicational spectrum of the myocutaneous vastus lateralis flap, which is often too voluminous for intraoral application, primary thinning of the fat and muscle component of this microsurgical transplant was performed in 14 patients. The surgical technique includes subfascial localization of at least one myocutaneous perforating vessel of the lateral circumflex femoral artery and its dissection through the fascia, muscles and fatty tissue up to the skin. The epifascial fatty tissue is completely removed except for a 1-2 cm wide cuff around the perforating vessel. The thinning technique was used for covering 10 intraoral and 4 extraoral defects and enabled the raising of skin flaps with a thickness of 4 mm even in obese patients. The vessel pedicle length of the thinned flaps was between 12 and 16 cm; flap size varied between 4x5 and 9x15 cm and the donor sites were directly closed. In one case, there was a partial necrosis (20%), but the other flaps healed without complications. The described method allows the raising of thick myocutaneous as well as thin skin flaps from the same donor region. PMID- 11030399 TI - Combined treatment of adenoid cystic carcinoma of the salivary glands. AB - A retrospective review of the experience of a single surgeon over a 20-year period is presented. Fifteen patients with primary adenoid cystic carcinoma of the major (6) or minor (9) salivary glands underwent combined treatment with wide local excision and radical postoperative radiotherapy. The actuarial survival is 100% at 5 years and 62% at 10 and 15 years. The disease-specific survival is 100% at 5 years and 86% at 10 and 15 years. There were no loco-regional recurrences. Wide local excision and radical postoperative radiotherapy seems to be an effective treatment for adenoid cystic carcinoma of the salivary glands. PMID- 11030400 TI - Three-phase bone scintigraphy and viability of vascularized bone grafts for mandibular reconstruction. AB - Three-phase bone scintigraphy was undertaken to check the anastomotic patency and monitor the viability of vascularized bone grafts. Ten consecutive patients who underwent vascularized bone grafting of the mandible were reviewed. A successful clinical outcome was achieved in 8 patients. The graft failed in 2 patients. In this series, 3-phase bone scintigraphy of radiolabeled (99m)Tc-methylene diphosphonate was performed at 7 days, and at 1, 3, 6, and 12 months after reconstruction. Assessments made using 3-phase bone images were compared with the clinical findings. The clinical outcome of the cases presented in our series correlated extremely well with 3-phase bone images. Three-phase bone scintigraphy is a useful method for the assessment of patency and viability of vascularized bone grafts. The use of this method can be very helpful in assessing the anastomotic patency and viability of a graft which for clinical reasons is suspected of being non-viable. PMID- 11030402 TI - The effect of an intra-articular bone fragment in the genesis of temporomandibular joint ankylosis. AB - The aim of this study was to examine the effect of an intra-articular bone fragment on the genesis of temporomandibular joint (TMJ) ankylosis. Twenty-two adult sheep had a unilateral operation of removal of the temporal and condylar articular surfaces, plus discectomy with or without insertion of an intra articular bone fragment. Functionally, radiologically, macroscopically and histologically, in the group with the intra-articular bone fragment, the joints showed more advanced ankylosis both at one and three months after the operation, as compared to the group with no intra-articular bone. This study shows that the bone fragment increases the extent of ankylosis in the sheep model. The clinical significance of the various experimental findings of this study and others in the same series is presented. PMID- 11030401 TI - Neoadjuvant, hyperfractionated irradiation induces apoptosis and decreases proliferation in squamous cell cancer of the oral cavity. AB - Twenty-eight patients with squamous cell carcinoma of the oral cavity were treated with a total dose of 20 Gy. Tissue samples for immunohistochemistry were taken at the time of diagnostic biopsy and at surgery after radiotherapy (RX). For detection of proliferating cells, the immunoperoxidase reaction with Ki67 was performed. Apoptotic cells were detected by the TdT-mediated biotin dUTP nick end labeling (TUNEL) method. RX reduced proliferation in 27 patients, only in one case did the proliferation index (PI) increase. Delta PI (PI before RX PI following RX) amounted to 4.11% (SD=3.2%; P<0.0001). The apoptotic index (AI) increased significantly subsequent to neoadjuvant RX. Delta AI (AI after RX--AI before RX) measured 1.82% (SD=0.9; P<0.001). These data indicate that RX of patients suffering from squamous cell carcinoma of the oral cavity with a dosis of 20 Gy induces apoptosis and inhibits proliferation of tumor cells. PMID- 11030403 TI - CD44 standard form (CD44H) expression and distribution in dysfunctional human temporomandibular joint discs. AB - The expression pattern of the cell adhesion molecule CD44 standard form (CD44H) in dysfunctional human temporomandibular joint (TMJ) discs was studied immunohistochemically and compared with normal disc pattern in order to evaluate the expression of this adhesion molecule and correlate it to histopathological changes. Immunohistochemistry with anti-CD44H antibodies was performed on paraffin sections of pathological and normal discs. In normal TMJ discs, a moderate immunolabelling with anti-CD44H antibodies was detectable in fibroblastlike cells, in the few fibrochondrocytes and in chondrocytelike cells. In dysfunctional discs, the staining pattern and intensity varied according to the histopathological findings of the specimens. The TMJ discs showing abnormal collagen arrangement or fragmentation of collagen fibres presented overall the same immunolabelling pattern of normal discs. In the discs showing areas of fibrocartilaginous metaplasia, CD44H expression was upregulated in fibrochondrocytes and fibroblastlike cells, especially around the chondroid tissue. Overall, these results suggest that CD44H mediates the binding of some ECM proteins in TMJ disc cells. The up-regulation of CD44H observed in some dysfunctional TMJ discs seems to indicate a prevention of apoptosis in fibroblastlike cells and an important role in phenotypical change of fibrochondrocytes into chondroblastlike cells, enabling the aggregation of chondroid tissue pericellular matrix components. PMID- 11030404 TI - Analysis of 70Kd heat shock protein expression in patients with internal derangement of the temporomandibular joint. AB - Recent studies have demonstrated that the expression of heat shock protein (HSP) was enhanced under stress in joint diseases, such as rheumatoid arthritis and osteoarthritis. The purpose of this study was to analyze the expression of 70Kd HSP in patients with internal derangement of the temporomandibular joint (TMJ) by immunohistochemical and enzyme-linked immunosorbent assay (ELISA) methods. For immunohistochemistry, 5 extirpated discs and 16 synovial biopsy specimens from patients with TMJ internal derangement and 2 extirpated discs from normal subjects were examined. For ELISA, synovial fluid from 11 patients with TMJ internal derangement and from 6 normal volunteers were investigated. The results showed that the 70Kd HSP staining intensity in chondrocytes around the damaged area of the articular discs from patients with TMJ internal derangement was higher than that in chondrocytes in control specimens. In addition, 70Kd HSP expression in synovial fluid from patients with TMJ internal derangement was slightly higher than that in normal subjects. These findings suggest that elevated 70Kd HSP expression is related to the progression of TMJ internal derangement. PMID- 11030405 TI - Survey of Australian and New Zealand Oral and Maxillofacial Surgery trainees and recent specialists--education and experience. AB - This study examined the demographic characteristics, educational background and attitudes toward training of all recent OMS specialists and all current trainees in Australia and New Zealand in 1996. The early nineties is a key period as it marks the transition from an essentially dentally based speciality (85% dental degree in 1990); to recent specialists with an increasing number with both medical and dental degrees (33% dual degree 1990-1996); to predominantly dual degree training (84.4% dual in 1996). Current trainees had more extensive experience in pathology, preprosthetic and reconstructive surgery. They were also strongly critical of the length and cost of training. PMID- 11030406 TI - Infotech corner. PMID- 11030407 TI - Molecular genetic advances in tuberous sclerosis. AB - Over the past decade, there has been considerable progress in understanding the molecular genetics of tuberous sclerosis, a disorder characterised by hamartomatous growths in numerous organs. We review this progress, from cloning and characterising TSC1 and TSC2, the genes responsible for the disorder, through to gaining insights into the functions of their protein products hamartin and tuberin, and the identification and engineering of animal models. We also present the first comprehensive compilation and analysis of all reported TSC1 and TSC2 mutations, consider their diagnostic implications and review genotype/phenotype relationships. PMID- 11030408 TI - Autosomal dominant polycystic kidney disease: molecular genetics and molecular pathogenesis. AB - Mutations in three different genes, PKD1, PKD2 and PKD3, can cause a very similar clinical picture of the autosomal dominant form of polycystic kidney disease (ADPKD). Apparently, mutations in the PKD3 gene, which is still unmapped, are very rare, whereas PKD1 defects account for about 85% of cases. Although ADPKD is a frequent monogenic disorder affecting approximately 1:1000 individuals in the Caucasian population, progress in understanding its pathology was somewhat slow until relatively recently when the PKD1 and PKD2 genes were mapped and cloned. They are both large, being approximately 52 kb and 68 kb in length respectively, and in addition, PKD1 is fairly complex, thus complicating mutation detection. The gene products, polycystin-1 and polycystin-2, are trans-membranous glycoproteins and are considered to be involved in signalling pathways, in cooperation with additional partners. Immunostaining studies in both humans and mice have revealed information regarding the localization of polycystins and their role in the development and maintenance of nephrons. Recent experimentation from various laboratories has shown that loss of heterozygosity and acquired somatic second hits may account, at least partly, for the inter- and intrafamilial phenotypic heterogeneity of the disease, while at the same time, the existence of other modifying loci is also hypothesized. The two-hit hypothesis is admittedly a very attractive one in that it can explain many of the features of the disease, whereas recent data regarding a trains-heterozygous model for cystogenesis adds to the complexity of the molecular mechanisms that can lead to pathogenesis. PMID- 11030409 TI - Genomic structure of a copy of the human TPTE gene which encompasses 87 kb on the short arm of chromosome 21. AB - The testis-expressed human TPTE is a putative transmembrane tyrosine phosphatase, probably involved in signal transduction pathways of the endocrine and/or the spermatogenetic function of the testis. TPTE was mapped to the pericentromeric region of human chromosomes 21 and 13, and to chromosomes 15, 22, and Y. It is unknown which of the TPTE copies are transcribed, contain intronic sequences, and/or have open reading frames. Here, in silico analysis of the genomic sequence of human chromosome 21 allowed the determination of the genomic structure of a copy of the TPTE gene. This copy consists of 24 exons and spans approximately 87 kb. The mapping position of this copy of TPTE on the short arm of chromosome 21 was confirmed by FISH using the BAC 15L0C0 clone as a probe that contains almost the entire TPTE gene. This is the first description of the genomic sequence of a non-RNR gene on the short arm of human acrocentric chromosomes. PMID- 11030410 TI - Genetic background of apparently idiopathic sporadic cerebellar ataxia. AB - Disease-causing mutations have been identified in various entities of autosomal dominant ataxia and in Friedreich's ataxia. However, no molecular pathogenic factor is known to cause idiopathic cerebellar ataxias. We investigated the CAG/CTG trinucleotide repeats causing spinocerebellar ataxia types 1, 2, 3, 6, 7, 8 and 12, and the GAA repeat of the frataxin gene in 124 patients apparently suffering from idiopathic sporadic ataxia, including 20 patients with the clinical diagnosis of multiple system atrophy. Patients with a positive family history, a typical Friedreich phenotype, or symptomatic ataxia were excluded. Genetic analyses uncovered the most common Friedreich mutation in 10 patients with an age at onset between 13 and 36 years. The SCA6 mutation was present in nine patients with disease onset between 47 and 68 years of age. The CTG repeat associated with SCA8 was expanded in three patients. One patient had SCA2 attributable to a de novo mutation from a paternally transmitted, intermediate allele. We did not identify the SCA1, SCA3, SCA7 or SCA12 mutation in idiopathic sporadic ataxia patients. No trinucleotide repeat expansion was detected in the MSA subgroup. This study has revealed the genetic basis in 19% of apparently idiopathic ataxia patients. SCA6 is the most frequent mutation in late onset cerebellar ataxia. The frataxin trinucleotide expansion should be investigated in all sporadic ataxia patients with onset before age 40, even when the phenotype is atypical for Friedreich's ataxia. PMID- 11030411 TI - Association of a variant in exon 31 of the sulfonylurea receptor 1 (SUR1) gene with type 2 diabetes mellitus in French Caucasians. AB - The sulfonylurea receptor (SUR1) of the pancreatic beta-cell ATP-sensitive potassium channel plays a key role in glucose-induced insulin secretion. The A allele of a single nucleotide polymorphism (SNP) in exon 31 of the SUR1 gene (AGG ->AGA; Arg1273Arg) has previously been shown to be associated with hyperinsulinemia in nondiabetic Mexican-American subjects. Here, we have investigated the association of this SNP with type 2 diabetes mellitus (T2DM) in French Caucasian subjects. We have observed an increased frequency of the A allele (37.1% vs 27.6%, P=0.0048; odds ratio 1.54), of the AA genotype (15.7% vs 9.8%; P=0.025), and of the combined AA/AG genotypes (58.5% vs 45.5%, P=0.0098; odds ratio 1.69) in patients compared with controls. This association is stronger in the subgroup of patients with age of diagnosis of diabetes equal to or less than 45 years: A allele 43.2% (P=0.0003 compared with controls; odds ratio 1.99), AA genotype 21.4% (P=0.0032), and combined AA/AG genotypes 65.1% (P=0.0022; odds ratio 2.23). Unexpectedly, the G allele is strongly associated with arterial hypertension in obese diabetic subjects (GG vs AA odds ratio 19.97). In conclusion, we have observed an association of an SNP in exon 31 of the SUR1 gene with T2DM. These data reinforce the hypothesis that insulin secretion defects in T2DM might be at least partially related to allelic variations in the SUR1 gene. PMID- 11030412 TI - Mutations in short stature homeobox containing gene (SHOX) in dyschondrosteosis but not in hypochondroplasia. AB - Dyschondrosteosis (DCO) and hypochondroplasia (HCH) are common skeletal dysplasias characterized by disproportionate short stature. The diagnosis of these conditions might be difficult to establish especially in early childhood. Point mutations and deletions of the short stature homeobox containing gene (SHOX) are detected in DCO and idiopathic short stature with some rhizomelic body disproportion, whereas mutations in the fibroblast growth factor receptor 3 (FGFR3) gene are found in 40-70% of HCH cases. In this study, we performed mutational analysis of the coding region of the SHOX gene in five DCO and 18 HCH patients, all of whom tested negative for the known HCH-associated FGFR3 mutations. The polymorphic CA-repeat analysis, direct sequencing and Southern blotting were used for detection of deletions and point mutations. The auxological and radiological phenotype of these patients was carefully determined. Three novel mutations in DCO patients were found: (1) a deletion of one base (de1272G) (according to GenBank accession nos. Y11536, Y11535), resulting in a premature stop codon at position 75 of the amino acid sequence; (2) the transversion C485G resulting in the substitution Leu132Val; and (3) the transversion G549T causing an Arg153Leu substitution. These substitutions segregate with the DCO phenotype and affect evolutionarily conserved homeodomain residues, based on a comparison of homeobox containing proteins in 13 species. Moreover, these changes were not found in 80 unrelated, unaffected individuals. This strongly suggests that these mutations are pathogenic. The phenotype of our patients with DCO and HCH varied from mild to severe shortness and body disproportion. These results further support clinical and genetic heterogeneity of dyschondrosteosis and hypochondroplasia. PMID- 11030413 TI - Porphobilinogen deaminase gene in African and Afro-Caribbean ethnic groups: mutations causing acute intermittent porphyria and specific intragenic polymorphisms. AB - Acute intermittent porphyria (AIP), the most common acute hepatic porphyria, is a low-penetrant autosomal dominant disorder caused by mutations in the porphobilinogen deaminase (PBGD) or hydroxymethylbilane synthase (HMBS) gene. Although AIP has been identified in all the main ethnic groups, little is known about PBGD gene defects in Africans, Afro-Caribbean and Afro-Americans. We have carried out PBGD gene screening among seven unrelated AIP families and 98 controls belonging to the Afro-Caribbean (French West Indies) and the sub-Saharan African (Morocco, Algeria, Cameroon, Mali, and Burkina Faso) populations. Using denaturing-gradient gel electrophoresis (DGGE) and direct sequencing we characterized six different mutations, including four novel, from the seven AIP families: three splicing defects (IVS 5+2 Ins G; IVS 7+1 G to A in two families; IVS 10-1 G to T); a small deletion (1004 Del G); and two missense mutations (R116 W; A270G). The allele frequencies of the 14 polymorphic sites, previously known in the normal Caucasian population, were similar in Africans and Afro-Caribbean control populations. Interestingly, two common new intragenic polymorphic sites, close to intron/junction boundaries, were identified only in blacks: 1) in intron 2, a single base-pair G deletion at position 3167 (G:0.88; delG:0.12); 2) in intron 10, a A/G dimorphism at position 7052 (A:0.56; G:0.44). These two single nucleotide polymorphisms (SNPs) were never encountered in 750 unrelated Caucasian subjects. The allele frequency distributions of populations within black ethnic groups (Africans and Afro-Caribbean) are similar. This study highlights differences both in PBGD gene mutations causing AIP and in SNPs between white and black peoples; the allele frequencies provided contribute to a better knowledge of the variability of these markers among the major population groups, especially in sub-Saharan West African and Afro-Caribbean populations. PMID- 11030414 TI - Immature end-plates and utrophin deficiency in congenital myasthenic syndrome caused by epsilon-AChR subunit truncating mutations. AB - Congenital myasthenic syndromes (CMS) are inborn disorders due to presynaptic, synaptic, or postsynaptic defects of neuromuscular transmission. Some previously described kinships with typical signs of CMS showed a marked deficiency of acetylcholine receptors (AChR) and utrophin at the neuromuscular junctions. Additionally, the end-plate ultrastructure was immature, with reduced enfolding of the postsynaptic membrane. In two such families, we found truncating mutations of the epsilon-AChR subunit. In family 1, both affected siblings were heteroallelic for a epsilon911delT and a epsilonIVS4+1G-->A mutation within the AChR epsilon-subunit gene (CHRNE). In the affected member of family 2, a epsilon1030delC mutation and a previously described epsilonR64X mutation were found. These deleterious epsilonAChR mutations not only result in AChR deficiency, but also affect end-plate maturation, including the formation of secondary synaptic clefts during ontogenesis. PMID- 11030415 TI - Characterization of TH1 and CTSZ, two non-imprinted genes downstream of GNAS1 in chromosome 20q13. AB - The clustering and coordinate regulation of many imprinted genes justifies positional searches for imprinted genes adjacent to known ones. We recently characterized a locus on 20q13, containing GNAS1, which has a highly complex imprinted expression pattern. In a search for neighbouring genes, we have now characterized a new gene, TH1, downstream of GNAS1. TH1 and GNAS1 are separated by more than 70 kb consisting largely of interspersed repetitive DNA. TH1 is the homologue of a gene that, in Drosophila, lies adjacent to the DNA repair gene mei 41. We have determined the full-length structures of human, mouse and Drosophila TH1. Though of unknown function, TH1 is highly conserved and widely expressed. Nonetheless, there is no similar Caenorhabditis elegans protein. We have also determined the complete genomic structures of human and Drosophila TH1. The Drosophila gene has five exons spanning 2.6 kb. The last three introns have precise equivalents in the human gene, which has 15 exons spanning 14 kb and is transcribed away from GNAS1. Using a single-nucleotide polymorphism in the 3' untranslated region, we have demonstrated biallelic TH1 expression in human fetal tissues, suggesting that, unlike GNAS1, TH1 is probably not imprinted. Immediately downstream of TH1 lies CTSZ, encoding the recently described cysteine protease, cathepsin Z. We have also elucidated the genomic structure of this gene; it has six exons spanning 12 kb and is oriented tail-to-tail with TH1, only 70 bp separating their polyadenylation sites. A polymorphism was again identified within the CTSZ 3' untranslated region and used to demonstrate biallelic expression in fetal tissues. PMID- 11030416 TI - Identification of missense, nonsense, and deletion mutations in the GRHPR gene in patients with primary hyperoxaluria type II (PH2). AB - Primary hyperoxaluria type II (PH2) is a rare disease characterized by the absence of an enzyme with glyoxylate reductase, hydroxypyruvate reductase, and D glycerate dehydrogenase activities. The gene encoding this enzyme (GRHPR) has been characterized, and a single mutation has been detected in four PH2 patients. In this report, we have identified five novel mutations. One nonsense mutation (C295T) results in a premature stop codon at codon 99. A 4-bp deletion mutation has been found in the 5' consensus splice site of intron D, resulting in a predicted splicing error. Three missense mutations have been detected, including a missense transversion (T965G) in exon 9 (Met322Arg), a missense transition (G494A) in the putative co-factor binding site in exon 6 (Gly165Asp), and a substitution of an adenosine for a guanine in the 3' splice site of intron G. The functional consequences of the missense transversion and transition mutations have been investigated by transfection of cDNA encoding the mutated protein into COS cells. Cells transfected with either mutant construct have no enzymatic activity, a finding that is not significantly different from the control (empty) vector (P<0.05). These results further confirm that mutations in the GRHPR gene form the genetic basis of PH2. Ten of the 11 patients that we have genotyped are homozygous for one of the six mutations identified to date. Because of this high proportion of homozygotes, we have used microsatellite markers in close linkage with the GRHPR gene to investigate the possibility that the patients are the offspring of related individuals. Our data suggest that two thirds of our patients are the offspring of either closely or distantly related persons. Furthermore, genotyping has revealed the possible presence of a founder effect for the two most common mutations and the location of the gene near the marker D9S1874. PMID- 11030417 TI - Prevalence of BRCA1 and BRCA2 mutations among clinic-based African American families with breast cancer. AB - To define the prevalence and relative contributions of BRCA1 and BRCA2 mutations among African American families with breast cancer, we analyzed 28 DNA samples from patients identified through two oncology clinics. The entire coding regions of BRCA1 and BRCA2 were screened by protein truncation test, heteroduplex analysis, or single-stranded conformation polymorphism followed by DNA sequencing of variant bands. Deleterious protein-truncating BRCA1 and BRCA2 mutations were identified in five patients or 18% of the entire cohort. Only 8% (1 of 13) of women with a family history of breast cancer, but no ovarian cancer, had mutations. The mutation rates were higher for women from families with a history of breast cancer and at least one ovarian cancer (three of six, 50%). One woman with a family history of undocumented cancers was also found to carry a deleterious mutation in BRCA2. The spectrum of mutations was unique in that one novel BRCA1 mutation (1625del5) and three novel BRCA2 mutations (1536del4, 6696delTC, and 7795delCT) were identified. No recurrent mutations were identified in this cohort, although one BRCA2 (2816insA) mutation had been previously reported. In addition, two BRCA1 and four BRCA2 missense mutations of unknown significance were identified, one of which was novel. Taken together with our previous report on recurrent mutations seen in unrelated families, we conclude that African Americans have a unique mutation spectrum in BRCA1 and BRCA2 genes, but recurrent mutations are likely to be more widely dispersed and therefore not readily identifiable in this population. PMID- 11030418 TI - Protein truncating BRCA1 and BRCA2 mutations in African women with pre-menopausal breast cancer. PMID- 11030419 TI - Detection of the fragile X syndrome protein for the evaluation of FMR1 intermediate alleles. AB - Molecular screening programs in mentally retarded individuals have been performed in several populations worldwide. One finding has been an excess of FMR1 intermediate alleles in a population with learning difficulties. However, other published reports with similar characteristics did not corroborate those previous results. In order to contribute additional data from our population, we studied 563 patients affected with nonspecific mental retardation (MRX) that did not present a CGG expansion in the FMR1 gene and 208 individuals as a control population. Forty MRX patients presented alleles within the intermediate range. Among them, one case showed a pattern of expression of the FMR1 protein (FMRP) concordant with a fragile X syndrome case with an intermediate allele/full mutation mosaicism, although it was not detected by Southern blot analysis. Statistical analysis was performed again showing no statistically significant difference regarding the intermediate allele frequency in the MRX and control populations. This finding is in agreement with the hypothesis that the incidence of intermediate FMR1 alleles in MRX populations does not seem to be higher than in control populations, and it emphasizes the importance of FMRP detection as a diagnostic tool for fragile X syndrome. PMID- 11030420 TI - Genetic association studies of bronchial asthma--a need for Bonferroni correction? PMID- 11030421 TI - The structural properties of plant peroxisomes and their metabolic significance. AB - Plant peroxisomes can be isolated by Percoll density gradient centrifugation at high purity and metabolic competence as well as in relatively large quantities. According to biochemical and electrophysiological analyses, plant peroxisomes have recently been shown to differ from other cell organelles in essential structural properties. Unlike mitochondria or plastids, compartmentalization of plant peroxisomal metabolism is in major parts not caused by a boundary function of the membrane but is primarily due to the specific structure of the protein matrix. The enzymes of the photorespiratory C2 cycle of leaf peroxisomes are arranged as multienzyme complexes that allow efficient metabolic channelling with high flux rates and minimum leakage of reactive oxygen species from the organelle. Transfer of metabolites, such as carboxylates, proceeds across the peroxisomal membrane via a porin-like channel, which represents a relatively unspecific but highly efficient transport system. Because all variants of peroxisomes, which all contain only a single boundary membrane, are confronted with the task of transporting a large group of metabolites while preventing the escape of reactive intermediates, it is reasonable to speculate that the unique compartmentalization feature of leaf peroxisomes also applies to peroxisomes from fungi and mammals. PMID- 11030422 TI - Active oxygen species as mediators of plant immunity: three case studies. AB - A burst of active oxygen species (AOS) is known to be involved in local cell death as part of plant defence against pathogens. It is, however, under dispute to what extent AOS can induce pathogen resistance and immunity throughout the plant. Three experimental strategies that reveal a primary role for AOS and a surprisingly low chemical and spatial specificity are now described for tobacco and Arabidopsis thaliana plants. Ozone is a gaseous AOS that was applied to non transgenic plants. Hydrogen peroxide or singlet oxygen are AOS that were induced by high-light treatment of transgenic plants that contained antisense constructs inhibiting catalase activity or chlorophyll biosynthetic enzymes. In all cases, activated oxygen species, cellular lesions, ethylene and salicylic acid, and components of major plant defence systems (systemic acquired resistance, hypersensitive response) were induced, as was resistance towards pathogens (tobacco mosaic virus, Pseudomonas syringae or Peronospora parasitica). It is concluded that active oxygen species can act as mediators of plant immunity so that new non-pesticidal plant protection strategies could be developed. PMID- 11030423 TI - Post-transcriptional regulation of phenylalanine ammonia-lyase expression in tobacco following recovery from gene silencing. AB - Introduction of a bean phenylalanine ammonia-lyase (PAL) transgene into tobacco plants results in epigenetic post-transcriptional gene silencing which is unstable, such that after self-pollination first generation progeny may become PAL over-expressors. The change from gene silencing to PAL over-expression is accompanied by a loss of cytosine methylation of the PAL transgene and reduced methylation of the endogenous tobacco PAL2 gene, but not the PAL1 gene. These changes are associated with the appearance of high levels of bean PAL and tobacco PAL2 transcripts in the total RNA fraction from PAL over-expressing plants. However, tobacco PAL2 transcripts are inefficiently recruited into polysomes, and tobacco PAL2 protein is not detected in leaves of PAL over-expressing or wild type lines. Thus, in spite of the post-transcriptionally controlled increase in tobacco PAL2 transcripts in PAL over-expressors, the increased PAL activity is primarily the result of the increase in bean PAL transcripts and corresponding enzymatic activity. These results reveal a complex cross-talk between expression of the PAL transgene and the corresponding endogenous PAL genes at the levels of transcription, transcript stability and polysomal recruitment during sense transgene-mediated silencing and subsequent over-expresson of PAL in tobacco. PMID- 11030424 TI - Two differentially regulated class II chitinases from parsley. AB - Two distinct cDNA clones, PcCHI1 and PcCHI2, with high sequence similarity to plant chitinases were isolated from parsley (Petroselinum crispum), expressed in Escherichia coli, and the encoded proteins functionally identified as endochitinases. Different expression patterns of the corresponding mRNAs and proteins in infected and uninfected parsley plants indicated distinct roles of the two isoforms in both pathogen defense and plant development. Infection of parsley leaf buds with Phytophthora sojae resulted in the rapid, transient and highly localized accumulation of PcCHI1 mRNA and protein around infection sites, whereas PcCHI2 mRNA and protein were systemically induced at later infection stages. Similar differences in the timing of induction were observed in elicitor treated, suspension-cultured parsley cells. In uninfected plants, PcCHI1 mRNA was particularly abundant in the transmitting tract of healthy flowers, suggesting a role in the constitutive protection of susceptible transmitting tissue of the style against pathogen ingress and/or in the fertilization process, possibly by affecting pollen tube growth. Localization of PcCHI2 mRNA and protein in the parenchymatic collenchyme of young pedicels may indicate a function in the constitutive protection of this tissue. In addition to such distinct roles of PcCHI1 and PcCHI2 in preformed and induced pathogen defense, both chitinases may have endogenous regulatory functions in plant development. PMID- 11030425 TI - IAA-synthase, an enzyme complex from Arabidopsis thaliana catalyzing the formation of indole-3-acetic acid from (S)-tryptophan. AB - An enzyme complex was isolated from Arabidopsis thaliana that catalyzes the entire pathway of biosynthesis of the major plant growth hormone, indole-3-acetic acid (IAA), from (S)-tryptophan. The 160-180 kDa, soluble complex catalyzes a strictly O2-dependent reaction which requires no further added factors and is stereospecific for the substrate (S)-tryptophan (app. Km = 120 microM). H2(18)O labeling proved that both oxygen atoms of IAA were delivered via H2O. This, as well as immunological evidence for the presence of a nitrilase-like protein in the complex, suggests the reaction to proceed via the intermediate indole-3 acetonitrile. IAA-synthase forms a tight metabolite channel committed to IAA production and occurs in shoots, roots and cell cultures of A. thaliana. PMID- 11030426 TI - Topology studies of the chloroplast protein import channel Toc75. AB - A major goal in understanding protein transport across membranes is the investigation of the structure and regulation of the translocon subunits. We analysed Toc75, a pore-forming subunit of the translocon of the outer envelope of chloroplasts. Toc75 was overexpressed and reconstituted into liposomes. Immunoprecipitation of liposome-reconstituted Toc75 indicates an N(in)-C(in) orientation of Toc75. Limited proteolytic digestion of Toc75 present in outer envelope vesicles with specific proteases combined with amino acid sequencing was used to study the topology of Toc75. Finally, computer modelling based on known protein structures indicates that Toc75 traverses the outer envelope with 16 amphiphilic beta sheets and the topology model is presented. PMID- 11030427 TI - Plant methionine synthase: new insights into properties and expression. AB - We investigated the enzyme methionine synthase (MSY) in Catharanthus roseus. The properties were characterized with purified protein isolated either from plant cell cultures or after heterologous expression in Escherichia coli. The protein was a monomer and accepted both the triglutamate (CH3-H4PteGlu3, apparent Km = 80 microM) and the monoglutamate (CH3-H4PteGlu1, apparent Km = 350 microM) of methyl 5,6,7,8-tetrahydropteroate as methyl donor, with a ratio of approximately 90:1 in favor of the triglutamate. Both activities required inorganic phosphate, but with different kinetics, and both were dependent on reducing agents. The activity required zinc, as shown by depletion and reconstitution experiments. Mg2+ had no effect on the activity. Two MSY isoforms purified from parsley cell cultures revealed the same properties as the C. roseus enzyme, however, the parsley proteins had no detectable activity with the monoglutamate substrate. The second part of the work compared the expression of the three enzymes of the methyl cycle (MSY, S-adenosyl-L-methionine synthetase, S-adenosyl-L-homocysteine hydrolase). In cell cultures, all three enzymes were present under all conditions investigated, with small changes at the protein level and more pronounced changes at the RNA level. Studies with seedlings revealed a low expression of all three enzymes in cotyledons, when compared to hypocotyls and radiculas. Immunohistochemical experiments indicated that MSY expression in cotyledons is cell-type specific, with the strongest signals detected in the upper epidermis. PMID- 11030428 TI - The hepta-beta-glucoside elicitor-binding proteins from legumes represent a putative receptor family. AB - The ability of legumes to recognize and respond to beta-glucan elicitors by synthesizing phytoalexins is consistent with the existence of a membrane-bound beta-glucan-binding site. Related proteins of approximately 75 kDa and the corresponding mRNAs were detected in various species of legumes which respond to beta-glucans. The cDNAs for the beta-glucan-binding proteins of bean and soybean were cloned. The deduced 75-kDa proteins are predominantly hydrophilic and constitute a unique class of glucan-binding proteins with no currently recognizable functional domains. Heterologous expression of the soybean beta glucan-binding protein in tomato cells resulted in the generation of a high affinity binding site for the elicitor-active hepta-beta-glucoside conjugate (Kd = 4.5 nM). Ligand competition experiments with the recombinant binding sites demonstrated similar ligand specificities when compared with soybean. In both soybean and transgenic tomato, membrane-bound, active forms of the glucan-binding proteins coexist with immunologically detectable, soluble but inactive forms of the proteins. Reconstitution of a soluble protein fraction into lipid vesicles regained beta-glucoside-binding activity but with lower affinity (Kd = 130 nM). We conclude that the beta-glucan elicitor receptors of legumes are composed of the 75 kDa glucan-binding proteins as the critical components for ligand recognition, and of an as yet unknown membrane anchor constituting the plasma membrane-associated receptor complex. PMID- 11030429 TI - Octadecanoid and jasmonate signaling in tomato (Lycopersicon esculentum Mill.) leaves: endogenous jasmonates do not induce jasmonate biosynthesis. AB - Jasmonates and their precursors, the octadecanoids, are signals in stress-induced alteration of gene expression. Several mRNAs coding for enzymes of jasmonic acid (JA) biosynthesis are up-regulated upon JA treatment or endogenous increase of the JA level. Here we investigated the positive feedback of endogenous JA on JA formation, as well as its beta-oxidation steps. JA-responsive gene expression was recorded in terms of proteinase inhibitor2 (pin2) mRNA accumulation. JA formed upon treatment of tomato (Lycopersicon esculentum cv. Moneymaker) leaves with JA derivatives carrying different lengths of the carboxylic acid side chain was quantified by gas chromatography-mass spectrometry (GC-MS). The data revealed that beta-oxidation of the side chain occurs up to a butyric acid moiety. The amount of JA formed from side-chain modified JA derivatives correlated with pin2 mRNA accumulation. JA derivatives with a carboxylic side chain of 3, 5 or 7 carbon atoms were unable to form JA and to express on pin2, whereas even-numbered derivatives were active. After treatment of tomato leaves with (10-(2)H)-(-)-12 oxophytoenoic acid, (4-(2)H)-(-)-JA and its methyl ester were formed and could be quantified separately from the endogenously nonlabeled JA pool by GC-MS analysis via isotopic discrimination. The level of 8 nmol per g fresh weight JA and its methyl ester originated exclusively from labeled 12-oxophytoenic acid. This and further data indicate that endogenous synthesis of the JA precursor 12 oxophytodienoic acid, as well as of JA and its methyl ester, are not induced in tomato leaves, suggesting that positive feedback in JA biosynthesis does not function in vivo. PMID- 11030430 TI - Metabolic activity decreases as an adaptive response to low internal oxygen in growing potato tubers. AB - Plants lack specialised organs and circulatory systems, and oxygen can fall to low concentrations in metabolically active, dense or bulky tissues. In animals that tolerate hypoxia or anoxia, low oxygen triggers an adaptive inhibition of respiration and metabolic activity. Growing potato tubers were used to investigate whether an analogous response exists in plants. Oxygen concentrations fall below 5% in the centre of growing potato tubers. This is accompanied by a decrease of the adenylate energy status, and alterations of metabolites that are indicative of a decreased rate of glycolysis. The response to low oxygen was investigated in more detail by incubating tissue discs from growing tubers for 2 hours at a range of oxygen concentrations. When oxygen was decreased in the range between 21% and 4% there was a partial inhibition of sucrose breakdown, glycolysis and respiration. The energy status of the adenine, guanine and uridine nucleotides decreased, but pyrophosphate levels remained high. The inhibition of sucrose breakdown and glycolysis was accompanied by a small increase of sucrose, fructose, glycerate-3-phosphate, phosphenolpyruvate, and pyruvate, a decrease of the acetyl-coenzymeA:coenzymeA ratio, and a small increase of isocitrate and 2 oxoglutarate. These results indicate that carbon fluxes are inhibited at several sites, but the primary site of action of low oxygen is probably in mitochondrial electron transport. Decreasing the oxygen concentration from 21% to 4% also resulted in a partial inhibition of sucrose uptake, a strong inhibition of amino acid synthesis, a decrease of the levels of cofactors including the adenine, guanine and uridine nucleotides and coenzymeA, and attenuated the wounding induced increase of respiration and invertase and phenylalanine lyase activity in tissue discs. Starch synthesis was maintained at high rates in low oxygen. Anoxia led to a diametrically opposed response, in which glycolysis rose 2-fold to support fermentation, starch synthesis was strongly inhibited, and the level of lactate and the lactate:pyruvate ratio and the triose-phosphate:glycerate-3 phosphate ratio increased dramatically. It is concluded that low oxygen triggers (i) a partial inhibition of respiration leading to a decrease of the cellular energy status and (ii) a parallel inhibition of a wide range of energy-consuming metabolic processes. These results have general implications for understanding the regulation of glycolysis, starch synthesis and other biosynthetic pathways in plants, and reveal a potential role for pyrophosphate in conserving energy and decreasing oxygen consumption. PMID- 11030431 TI - Structure-activity relationships of synthetic analogs of jasmonic acid and coronatine on induction of benzo[c]phenanthridine alkaloid accumulation in Eschscholzia californica cell cultures. AB - A facile test system based on the accumulation of benzo[c]phenanthridine alkaloids in Eschscholzia californica cell suspension culture (an indicator of defense gene activation) has been used to analyze a series of synthetic compounds for elicitor-like activity. Of the 200 jasmonic acid and coronatine analogs tested with this system, representative results obtained with 49 of them are presented here. The following can be summarized concerning structure-activity relationships: there is a large degree of plasticity allowed at the C-3 of jasmonic acid in the activation of defense genes. The carbonyl moiety is not strictly required, but exocyclic double bond character appears necessary. The pentenyl side chain at C-2 cannot tolerate bulky groups at the terminal carbon and still be biologically active. Substitutions to the C-1' position are tolerated if they can potentially undergo beta-oxidation. Either an alkanoic acid or methyl ester is required at C-1, or a side chain that can be shortened by beta oxidation or by peptidase hydrolysis. Coronatine and various derivatives thereof are not as effective as jasmonic acid, and derivatives in inducing benzo[c]phenanthridine alkaloid accumulation. Jasmonic acid rather than the octadecanoic precursors is therefore considered to be a likely signal transducer of defense gene activation in planta. PMID- 11030432 TI - Identification of the Arabidopsis thaliana flavonoid 3'-hydroxylase gene and functional expression of the encoded P450 enzyme. AB - The phenylpropanoid pathway results in the synthesis of thousands of compounds, including flavonoids like flavonols, anthocyanidins and tannins. In Arabidopsis thaliana, the lack of tannins in the seed coat (testa) causes the transparent testa (tt) phenotype. In the present study, we identified the gene responsible for the tt7 mutation. We show that TT7 encodes the enzyme flavonoid 3' hydroxylase (F3'H), and demonstrate that this P450-dependent monooxygenase has F3'H activity. The availability of the AtF3'H gene and promoter sequence will allow us to study the coregulation of a complete set of flavonol and anthocyanidin biosynthesis genes in A. thaliana, and makes in vitro synthesis of hydroxylated flavonoids more feasible. PMID- 11030433 TI - Gut bacteria may be involved in interactions between plants, herbivores and their predators: microbial biosynthesis of N-acylglutamine surfactants as elicitors of plant volatiles. AB - N-Acylamino acids are dominant and widespread constituents of insect oral secretions (regurgitants), serving the insect as biosurfactants in the digestive process. During feeding the conjugates may be introduced into damaged leaves and contribute there to the elicitation of plant defenses such as the induction of volatile biosynthesis. From gut segments of Spodoptera exigua, Mamestra brassicae and Agrotis segetum 23 bacterial strains were isolated, ten of which were able to synthesise typical lepidopteran N-acylamino acids from externally added precursors. Four strains, Providencia rettgeri, Ochrobactrum spec., Myroides odoratus and Acinetobacter sp. genospecies 11 were identified on the basis of their 16 S rDNA. The organisms displayed a very broad substrate tolerance, since fatty acids of different chain length and different degree of saturation were converted into N-acylamino acids. Moreover, most of the proteinogenic amino acids, but not glutamic and aspartic acid, were used as substrates. The dominant occurrence of fatty acids conjugated with glutamine may result from a preferred transport of glutamine from the hemolymph into the gut of the insects. The involvement of bacteria in the biosynthesis of compounds which play a pivotal role in the interaction of plants, herbivores and their predators adds a new trophic level to this complex network of interactions. Due to their short generation cycle and the ease of adaptation endosymbiontic bacteria may have an outstanding importance for the coevolution of plant-insect interactions. PMID- 11030434 TI - Different signaling pathways are involved in CCK(B) receptor-mediated MAP kinase activation in COS-7 cells. AB - Recently, the involvement of the MAP kinase ERK in mitogenic signaling of cholecystokininB (CCK(B)) receptors has been shown. However, the intracellular effector systems involved in this signaling pathway are poorly defined. In this study, we used COS-7 cells transiently transfected with the human CCK(B) receptor to investigate cholecystokinin-induced MAP kinase activation. CCK-8 induced activation of ERK2 which is associated with its phosphorylation and localization in the nucleus. The CCK-8-dependent ERK stimulation is sensitive to wortmannin an inhibitor of phosphoinositide 3-kinases (PI3Ks) indicating the involvement of PI3K activity. To identify the PI3K species involved in mitogenic signaling of the CCK(B) receptor several dominant-negative mutants of PI3K regulatory and catalytic subunits were transiently expressed. Surprisingly, different catalytically inactive mutants of the G protein-sensitive PI3Kgamma did not affect ERK stimulation induced by CCK, whereas a dominant-negative mutant of the regulatory p85 subunit induced significant inhibition of CCK-dependent ERK activity. These results indicate an involvement of PI3K class 1A species alpha, beta or/and delta in signal transduction via CCK(B) receptors. In addition, protein kinase C (PKC)-dependent signaling pathways contribute to CCK(B)-mediated MAP kinase signaling as shown by inhibition of CCK-8-induced ERK activation by the PKC inhibitor bisindolylmaleimide. PMID- 11030435 TI - Non-specific depurination activity of saporin-S6, a ribosome-inactivating protein, under acidic conditions. AB - Among five ribosome-inactivating proteins tested only saporin-S6 could efficiently release the adenine from adenosine 20 of the synthetic oligoribonucleotide (SRD RNA) mimic of the sarcin/ricin domain of rat 28S rRNA with a Km of 9 microM and a kcat of approximately 0.4 min(-1) at pH 7.6. The optimal pH for the depurination activity of saporin-S6 is 5.0. However, saporin S6 lost its site-specificity of depurination on SRD RNA around the optimal pH. The non-specific depurination activity of saporin-S6 was dependent on the enzyme concentration and pH conditions. These results are valuable to understand the diversity and the depurination mechanism of ribosome-inactivating proteins. PMID- 11030436 TI - Molecular evolution in the hypervariable regions of fetuin: comparison between human and African green monkey fetuin. AB - Sequences of fetuin cDNA and its deduced amino acid residues from the African green monkey cell line Vero were found to differ by 7.3% and 12.9%, respectively, from the corresponding human sequences. Most amino acid substitutions were clustered within a small segment of the third domain (D3). Calculations of nonsynonymous and synonymous nucleotide substitution rates suggest that this small segment was mutated under positive selection. cDNAs encoding alpha1 antitrypsin, beta-actin and the sequences of intron 4 of alpha1-antitrypsin gene in human liver and Vero cells were also investigated. The results substantiated the positive selection imposed on the D3 segment. PMID- 11030437 TI - Enzyme induction and inhibition by new antiepileptic drugs: a review of human studies. AB - The aim of this paper is to review a number of new antiepileptic agents (i.e. felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, tiagabine, topiramate, vigabatrin and zonisamide) for their inducing and/or inhibitory properties in humans, mainly considering the interactions where they are involved as the cause rather than the object of such interactions. Two aspects have been particularly taken into account: the changes or absence of changes in plasma/serum concentrations of concomitant drugs and the direct or indirect evidence of induction, inhibition or lack of effect on the six major human hepatic CYP isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4), as well as on other CYP isozymes or enzyme systems. Felbamate clearly affects the pharmacokinetics of a number of drugs, generally increasing but also decreasing their concentrations. It induces enzymes such as CYP3A4 and inhibits enzymes such as CYP2C19 and those of the beta-oxidation pathway. Topiramate is not devoid of potential interaction properties: it decreases the plasma concentrations of ethinylestradiol, induces CYP3A4 and inhibits CYP2C19. For oxcarbazepine, no inhibitory, only inductive effects have been observed thus far. Felbamate. topiramate and oxcarbazepine may induce the metabolism of steroidal oral contraceptives. In this respect, tiagabine has been studied at a rather low dose. Pharmacodynamic or pharmacokinetic interaction seems to exist between lamotrigine and carbamazepine. Lamotrigine appears to be a weak inducer of UGTs, whereas induction of CYP3A4 seems improbable as the compound does not change the concentrations of oral contraceptives or the urinary excretion of 6beta hydroxycortisol. Zonisamide has very peculiar pharmacokinetics and an extensive metabolism. Additional information on its enzyme inducing or inhibiting properties would be necessary, as data so far collected on its effect on the pharmacokinetics of other drugs are conflicting. Gabapentin, vigabatrin and in particular levetiracetam appear to be devoid of significant enzyme inducing or inhibiting properties. PMID- 11030438 TI - Combination treatment for chronic hepatitis C: what is the role of ribavirin? AB - Ribavirin in combination with interferon-alpha2b is the new standard for chronic hepatitis C (CHC) treatment. Although usually considered as an antiviral compound, this guanosine analogue shows some additional effects on the immune system that could largely contribute to its clinical efficacy in CHC. Numerous in vitro experiments demonstrate that ribavirin has a selective down-regulatory effect on TH2 cytokine release with, in some cases, a concomitant TH1 cytokine up regulation. In vivo, combination treatment of CHC patients was shown to induce a predominant TH1 response in isolated PBMCs, but also a reduction of peripheral TH2 response. Considering that: 1) a strong CD(4)+ helper T-cell response is associated with viral clearance in acutely infected patients; 2) a weak T-cell response to the viral antigens is common in chronic infected patients; 3) responding patients to combination treatment (but not non-responding patients) altered their cytokine profile under treatment, either to express IFN-gamma or to reduce pro-inflammatory mediators; it is highly presumed that ribavirine participates to restore an efficient T-cell response and to reduce the non specific inflammatory cytolytic activity during CHC combination treatment. PMID- 11030439 TI - Additivity of bupivacaine and morphine for peripheral analgesia in rats. AB - Infiltration of the surgical wound is a classical technique for post-operative analgesia. Recent studies have suggested that local anaesthetic may be combined with other drugs such as opioids. This study has evaluated, in rat, the infiltration with morphine, bupivacaine and their combination. In all groups, the two hind paws were injected with carrageenin. The left hind paw was used as control. The vocalisation threshold to paw pressure (VTPP) of both hind paws was evaluated 2 h after induction of carrageenin inflammation (baseline value), then every 10 min until the return to baseline value after injection of analgesic drugs. The development of oedema was evaluated in both hind paws by measurement of paw circumference (PC) before, then after, carrageenin injection. All analgesic drugs were injected in the right inflamed paw diluted in 0.2 mL of normal saline. The analgesic effect of bupivacaine (0.1, 0.25 and 0.5%), morphine (25, 50 and 100 microg) and their combination (bupivacaine 0.1%/morphine 20 microg, bupivacaine 0.2%/morphine 40 microg and bupivacaine 0.4%/morphine 80 microg) was tested. The effect of naloxone on morphine induced analgesia was tested. The interaction between bupivacaine and morphine was evaluated with an isobolographic analysis. Bupivacaine produced a dose-dependent antinociceptive effect. Morphine infiltration produced a peripheral, dose-dependent analgesic effect antagonised by naloxone. This analgesic effect of morphine was associated with an anti-inflammatory effect. The isobolographic analysis revealed only additivity between bupivacaine and morphine. The infiltration with morphine offers a peripheral analgesic effect which is additive with the effect of bupivacaine. An anti-inflammatory effect of morphine participates in this peripheral analgesic effect. PMID- 11030440 TI - Tramadol relieves thermal hyperalgesia in rats with chronic constriction injury of the sciatic nerve. AB - The present study was designed to test whether tramadol is effective in the control of neuropathic pain in rats. Chronic constriction injury (CCI) of the sciatic nerve was induced over the left hind limb in male Sprague-Dawley rats. Identical surgery was performed on the opposite side except that the sciatic nerve was not ligated (sham surgery). Paw withdrawal latency (PWL) to heat was tested for each hind paw 1 day before surgery and on the 4th day after surgery to ensure the development of thermal hyperalgesia. In the acute treatment groups, saline or tramadol was administered subcutaneously at doses of 10, 20 or 30 mg/kg, and PWLs were measured 30, 60, 90, 120, 150 and 180 min after treatment. In the semi-chronic treatment groups, continuous systemic administration of tramadol 40 mg/kg/day or saline for 7 days was provided at a uniform rate via osmotic mini pumps. Tramadol reversed PWL in a dose-dependent manner in the acute treatment groups. PWLs were significantly reversed at 2 days after tramadol infusion, and this effect was sustained throughout the remainder of the treatment period in comparison with the saline group. Tramadol also resulted in a decreased sensitivity to thermal stimulus on the sham limb both in acute and semi-chronic administration. We conclude that both acute and semi-chronic tramadol treatment relieves thermal hyperalgesia effectively in rats with CCI of the sciatic nerve. This indicates that tramadol shows promise as a potential treatment for relief of neuropathic pain in humans. PMID- 11030441 TI - Effect of acute atenolol on short-term blood pressure variability and baroreflex sensitivity in rats. AB - Beta-blocker therapy for hypertension or coronary artery disease is common, but there are a lot of controversies about its effects on short-term blood pressure variability and arterial baroreceptor reflexes. The aim of this study was to evaluate the effects of acute atenolol on baroreflex sensitivity (BRS) and on the spontaneous variability of systolic blood pressure (SBP) and RR intervals in conscious rats. Ten Wistar rats equipped with telemetry system were evaluated: 1) under control conditions; 2) after injection of saline; and 3) during beta1 adrenergic blockade by atenolol. Fast Fourier transform analysis was applied to RR intervals and SBP. Atenolol increased RR intervals significantly by 14% and the variation coefficient of the RR intervals by 31%. SBP was reduced significantly by 9%. In frequency domain, beta1-blockade in RR intervals increased very low frequency by 33% and the total power by 22% and decreased low frequency by 25%. The ratio of low to high frequency power decreased by 60%. Frequency domain variables in SBP were not significantly changed after beta1 adrenergic receptor blockade. BRS (gain alpha) was not significantly altered by beta-blockers. Acute atenolol decreased SBP and increased RR intervals with no change in BRS, indicating 'resetting' of baroreflex function. PMID- 11030442 TI - Characterization and mechanisms of the cardiovascular and haemodynamic alterations induced by scorpion venom in rats. AB - The scope of this work was to investigate the nature, chronology and mechanisms of the cardiovascular disorders induced by scorpion envenomation. Anaesthetized rats were instrumented for measurement of cardiac output (CO), renal (RBF) and muscular (HBF) blood flows (pulsed Doppler flowmetry), blood pressure, heart rate and dP/dt. Buthus occitanus venom (BO) was administered intravenously in the absence/presence of different pre-treatments. BO dose-dependently (150-300 microg/kg) increased blood pressure, dP/dt, total peripheral (TPR), renal (RVR) and muscular (HVR) vascular resistances, and decreased CO, RBF and HBF. Recovery occurred after 150 but not after 300 microg/kg. BO, 600 microg/kg, produced qualitatively similar effects but arrhythmias developed and mortality increased. Pre-treatment with phentolamine prevented the rises in TPR, RVR, HVR and blood pressure and the decreases in CO, RBF and HBF induced by BO, 300 microg/kg. Pre treatment with propranolol prevented the rise in dP/dt and the occurrence of arrhythmias and limited the rise in RVR and the drop in RBF induced by BO, 300 microg/kg. Phentolamine, propranolol and their combination also prevented BO, 600 microg/kg-induced mortality. Other pre-treatments (bosentan, losartan, diltiazem, mepyramine) were almost ineffective vs. BO effects. Finally, BO, 300 microg/kg, induced a 30-40-fold increase in plasma epinephrine and norepinephrine levels, but no change in plasma endothelin-1 levels. Thus in anaesthetized rats, the pattern of the cardiac and systemic and regional haemodynamic effects of BO is typically that of and results from catecholamine outpouring-induced alpha- and beta-adrenoceptor stimulation. PMID- 11030443 TI - Direct vascular actions of methyclothiazide and indapamide in aorta of spontaneously hypertensive rats. AB - In vitro experiments were designed to assess the inhibitory effect of the thiazide diuretics methyclothiazide (MCTZ), the hydrochlorothiazide (HCTZ), and the thiazide-related diuretic indapamide (IND) on contractile responses to norepinephrine (NE) and arginine vasopressin (AVP) of aortic rings from spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY). Changes in the tension of aortic ring preparations were measured isometrically. MCTZ (10(-4) M) induced endothelium-dependent inhibition of the vasoconstrictor responses to NE and AVP only in aortas from SHR, and the maximal vasoconstrictive effect of NE and AVP was decreased by 59 +/- 11% and 32.3 +/- 13%, respectively. Indapamide (10(-4) M) also induced endothelium-dependent inhibition of the contractile response to AVP in aortic rings from SHR, and the maximal vasoconstrictive effect of AVP was decreased by 33 +/- 5%. In contrast, HCTZ did not inhibit the contractile response to either NE or AVP, even at the highest concentration. This study provides evidence that methyclothiazide and indapamide inhibit the contractile response induced by norepinephrine and/or arginine vasopressin on SHR aortic preparations via an endothelium-dependent mechanism. PMID- 11030444 TI - Amiloride-sensitive pressure-induced myogenic contraction in rat cerebral artery. AB - The inhibitory action of amiloride on the pressure-induced contraction was assessed in isolated rat cerebral artery. The artery was mounted in an arteriograph, and the change in intracellular Ca2+ concentration ([Ca2+]i) and vessel diameter were simultaneously measured. The contractile response elicited by intraluminal pressurization was independent of endothelium, i.e. myogenic in nature, and abolished by nicardipine, a Ca2+ antagonist or by removal of extracellular Ca2+, and was potentiated by 25 mM KCl. Cyclopiazonic acid and thapsigargin, inhibitors of the Ca2+-ATPase pump of the sarcoplasmic reticulum, and a protein kinase C inhibitor calphostin C did not suppress the pressure induced contraction. Amiloride, a putative stretch-activated cation channel blocker, attenuated with an IC50 (50% inhibitory concentration) of about 3 microM the increase in [Ca2+]i and contractile activity in response to pressure, whereas the drug showed no apparent effect on the contraction produced by high KCl or 9,11-dideoxy-11alpha,9alpha-epoxymethano prostaglandin F2alpha (U46619). Furthermore, amiloride (100 microM) did not significantly affect intracellular pH in the artery. In spite of its multiple pharmacological actions, it seems possible that amiloride is a useful alternative tool at the cellular or tissue level to study the mechanotransduction mechanisms involved in the pressure induced contraction in rat cerebral artery. PMID- 11030445 TI - Effects of pimobendan, a new cardiotonic agent, on contractile responses in single skeletal muscle fibres of the frog. AB - Effects of a new cardiotonic agent, pimobendan, on contraction were investigated in single intact skeletal muscle fibres of the frog. Pimobendan increased twitch tension in a concentration-dependent manner regardless of the presence or absence of Ca2+ without any effect on tetanic tension, the resting membrane potential and the shape of the action potential. Pimobendan caused a further increase in twitch tension potentiated by caffeine (1 mM). Adenine, an inhibitor of Ca2+-induced Ca2+ release from the sarcoplasmic reticulum, inhibited twitch tension potentiated by caffeine but not by pimobendan, suggesting that twitch potentiation by pimobendan is not attributed to increases in Ca2+-induced Ca2+ release. Pimobendan failed to increase cAMP levels in the skeletal muscle, though forskolin significantly increased it without any effect on twitch tension. Contractile responses to high concentrations of caffeine and K+ were also potentiated by pimobendan. These results suggest that the potentiating effect of pimobendan on skeletal muscle contraction is mainly due to the increase in Ca2+ sensitivity to the contractile apparatus. PMID- 11030446 TI - Identification of opioid receptors in the sympathetic and parasympathetic nerves of guinea-pig atria. AB - The opioid receptor subtypes of autonomic nerves of guinea-pig atria were elucidated by monitoring the effects of selective opioid receptor agonists on the negative and positive inotropic responses associated with the stimulation of the parasympathetic and sympathetic nerves, respectively. The positive inotropic effect, evoked by electrical field stimulation (2 Hz) was strongly reduced by the selective OP2-opioid receptor agonists U-50488 and U-69593, but partly by the OP3 opioid receptor agonist morphine. This effect of U-50488 and U-69593 were reversed by the selective OP2-opioid receptor antagonist nor-BNI. The effect of morphine was partly reversed by naloxone, whereas OP1-opioid receptor agonists, BW373 U86 and DPDPE, were ineffective. On the other hand, the negative inotropic response to electrical field stimulation was not affected by opioid receptor agonists. These results suggest that the noradrenaline release from cardiac sympathetic nerves of guinea-pig could be modulated, mainly by the OP2-opioid receptor, however, the acetylcholine release from cardiac parasympathetic nerves is not modulated by opioid receptors. PMID- 11030447 TI - Interaction of extracellular albumin and intravenous anaesthetics, etomidate and propofol, on calcium signalling in rat airway smooth muscle cells. AB - It has been shown in vitro that general anaesthetics modify airway responsiveness via, at least partially, a direct inhibitory effect on calcium signalling in airway smooth muscle cells. However, in vivo, these anaesthetic compounds bind serum proteins. We have investigated the effect of exposure to extracellular albumin of freshly isolated airway smooth muscle cells on the propofol- and etomidate-induced inhibitory effect on calcium signalling. [Ca2+]i was measured by microspectrofluorimetry in rat isolated tracheal smooth muscle cells using the fluorescent dye indo-1. Propofol (3 x 10(-4) M) and etomidate (10(-4) M) were the lowest 'effective' concentrations that altered the [Ca2+]i response. This alteration consisted of a decrease in both the amplitude of the [Ca2+]i peak (from 358 +/- 13 nM to 65 +/- 15 and 108 +/- 27 nM for propofol and etomidate, respectively) and the percentage of responding cells (from 80% to 37 and 25% respectively) in response to the low concentration of ACh and a decrease in the Ca2+ oscillation frequency (from 9.9 +/- 0.3 min(-1) to 4.7 +/- 0.4 and 6.9 +/- 0.4 min(-1), respectively) in response to the high concentration of ACh. Increasing the concentration of albumin reduced the inhibitory effect of etomidate and propofol on the [Ca2+]i response to ACh. When extracellular albumin concentration was kept constant (20 g/L), increasing the concentration of etomidate by one log restored its inhibitory effect on the calcium signal. This study indicates that increasing the concentration of extracellular albumin reduces the inhibitory effect of intravenous anaesthetics on calcium signalling in airway smooth muscle cells. This report suggests that, in extrapolating in vitro dose-response relationships to those from in vivo conditions, the effect of the concentration of extracellular protein can be estimated. PMID- 11030448 TI - Differential transcriptional regulation of endothelin-1 by immunosuppressants FK506 and cyclosporin A. AB - Calcineurin antagonists FK506 and CsA, administered to treat organ allograft rejection, exert specific effects on renal vasoconstriction and nephrotoxicity, possibly due to endogenous vasoconstrictor release such as ET-1. We investigated contribution of FK506 and CsA on regulation of prepro ET-1 gene transcription in HUVEC. To conclude on transcriptional regulation, ET-1 mRNA levels were quantified by Northern blot analysis upon stimulation with calcineurin antagonists, and newly transcribed luciferase gene, placed under the control of the rat ET-1 promoter, was quantified by reporter gene assays, where luciferase activity reflects ET-1 promoter activation. Calcium fluorometry was employed to examine calcium dependency of ET-1 promoter-dependent gene transcription. Northern blot analysis shows differential induction of prepro ET-1 mRNA in favour of CsA over FK506. Likewise, luciferase assays demonstrate stronger ET-1 promoter dependent stimulation of the reporter gene by CsA than by FK506. Transcription of prepro ET-1 gene upon stimulation with both calcineurin antagonists is regulated by intracellular calcium levels. Lack of extra- or intracellular calcium prevents ET-1 promoter-dependent gene transcription and ET-1 mRNA induction. These observations demonstrate that calcineurin antagonists FK506 and CsA differ in quality to induce transcription of prepro ET-1 in HUVEC via calcium-dependent nuclear signalling events. To examine the contribution of ET-1 in nephrotoxicity upon CsA and FK506 immunosuppression the availability of endothelin receptor antagonists or endothelin converting enzyme inhibitors is required. PMID- 11030449 TI - Activity of ebastine (10 and 20 mg) and cetirizine at 24 hours of a steady state treatment in the skin of healthy volunteers. AB - We have compared the inhibitory effects of ebastine (10 mg), ebastine (20 mg) and cetirizine (10 mg) on histamine-induced wheal and flare skin reactions 24 h following a 6-day-long treatment. This was a double-blind, randomised, crossover, placebo-controlled study involving 24 healthy volunteers (18-65 years) with negative skin prick tests and the absence of specific IgEs to common allergens. Subjects were randomised to receive each of the following treatments once daily for 6 days: ebastine (10 mg), ebastine (20 mg), cetirizine (10 mg) or placebo with a washout period of 5 days. Twenty-four hours after the last dose of each treatment, histamine skin prick tests were performed (0, 0.5, 1, 2.5, 5, 10, 20, 50, 100 and 200 mg/mL), and wheal and flare responses were measured. All active treatments produced significant inhibition of the wheal responses compared to placebo (P < 0.001). Wheal response inhibition was significantly better with 20 mg of ebastine compared with 10 mg of ebastine and 10 mg of cetirizine. In a comparison to histamine concentrations required to produce a wheal surface area of 10 mm2, 20 mg of ebastine was also significantly better than ebastine 10 mg and cetirizine (P < 0.001), and 10 mg ebastine was significantly better than cetirizine (P < 0.05). Highly significant (P < 0.001) effects on the flare response were observed with each active treatment compared to placebo, with no difference between groups. The frequency of adverse events, primarily somnolence, was similar among the four treatment groups. Our results clearly indicate that ebastine, at either recommended dosage of 10 and 20 mg, and cetirizine produced significant inhibition of the histamine-induced wheal and flare reaction compared to placebo for up to 24 h. A superior efficacy of 20 mg of ebastine is observed compared with 10 mg of ebastine and 10 mg of cetirizine on the skin wheal response 24 h after the last dose of a 6-day-long treatment. This study clearly proves ebastine to be an effective, truly once-daily antihistamine. PMID- 11030450 TI - Pharmacokinetics of cyclophosphamide (CP) and 4-OH-CP/aldophosphamide in systemic vasculitis. AB - Cyclophosphamide given in association with corticosteroids has markedly improved the prognosis of systemic vasculitis. Little information has been reported on cyclophosphamide pharmacokinetics in these diseases and data evaluating its metabolite, 4-hydroxycyclophosphamide/aldophosphamide, pharmacokinetics and concentrations are lacking. Cyclophosphamide was administered as a 1-h intravenous infusion every 3 weeks for six cycles to ten vasculitis patients. Serum cyclophosphamide and 4-hydroxycyclophosphamide/aldophosphamide concentrations were assayed on the first cycle of the treatment by reversed-phase high-pressure liquid chromatography with ultraviolet detection. The mean (+/- SD) 4-hydroxycyclophosphamide/aldophosphamide and cyclophosphamide areas under the serum concentration-time curves were, respectively, 1.86 +/- 1.12 and 154.1 +/- 62.7 mg/L x h with a ratio of 1.30 +/- 0.76%. The mean maximum serum 4 hydroxycyclophosphamide/aldophosphamide was reached 2.3 h after cyclophosphamide administration. The mean (+/- SD) cyclophosphamide and 4 hydroxycyclophosphamide/aldophosphamide half-lives were, respectively, 5.5 +/- 3.1 and 7.6 +/- 2.3 h. The results are consistent with those obtained for cancer patients, in spite of a wide interpatient variability of concentrations and pharmacokinetic parameters. PMID- 11030451 TI - Post-traumatic amnesia after closed head injury: a review of the literature and some suggestions for further research. AB - Post-traumatic amnesia (PTA) is a transient sequela of closed head injury (CHI). The term PTA has been in clinical use for over half a century, and generally refers to the subacute phase of recovery immediately after unconsciousness following CHI. The duration of PTA predicts functional outcome after CHI, but its pathophysiological mechanism is not known. This paper compares current methods of determining the duration of PTA, summarizes reports on neuropsychological deficits in PTA, reviews available data that allow inferences about its mechanism, and suggests methods for further exploration of its pathophysiology. PMID- 11030452 TI - Acute behavioural disturbances related to imaging studies and outcome in mild-to moderate head injury. AB - The frequency of behavioural disturbances early after injury in relation to outcome was prospectively investigated in a series of 67 patients with mild-to moderate head injury (as defined by GCS on admission). In more than half of the patients, behavioural disturbances were observed. Restlessness occurred in 40% of patients, whereas agitation was seen in 19% of patients. In all patients, restlessness and agitation disappeared before resolution of PTA. In multiple regression analysis, restlessness and PTA were found to be separate factors in predicting outcome. On imaging studies, twice as many lesions were seen in patients with restlessness and agitation (81% compared to 39%), mainly localized in the frontotemporal region. In two thirds of patients with early behavioural disturbances, residual emotional and cognitive impairments were seen 1 year after injury. This study suggests that behavioural disturbances in the early phase after injury are related to frontotemporal lesions and lends support for the view of the existence of a separate profile of patient behaviour in mild-to-moderate head injury. PMID- 11030453 TI - Phonemic and semantic fluencies in traumatic brain injury patients with focal frontal lesions. AB - The purpose of this study was to investigate the effect of focal frontal lesions, identified by magnetic resonance imaging (MRI), in a group of traumatic brain injured (TBI) patients, in order to clarify the sensitivity of phonemic and semantic fluency tasks as tests of frontal lobe functioning. Thirteen TBI patients were included and matched with a normal control group of 26 subjects. Frontal lobe patients produced significantly fewer words than the control group in the phonemic fluency condition. Semantic performance correlated with lesion size, but phonemic performance did not, corroborating the idea that the two have different brain-based substrates. Although, as a group effect, frontal lesions impaired fluency, not all patients were impaired, even in the presence of large bilateral frontal lesions. PMID- 11030454 TI - Chronic neurobehavioural effects of mercury poisoning on a group of Zulu chemical workers. AB - PRIMARY OBJECTIVE: To assess the nature and severity of reported neurobehavioural symptoms of mercury poisoning, in a group of Zulu chemical workers (n = 16), employed by a mercury processing plant, exposed to neurotoxic levels of mercury, 5 years after exposure. RESEARCH DESIGN: A group-control design was adopted, where the exposed group was matched for age, sex, race, occupational and educational background. METHOD/PROCEDURES: Both groups were administered a specially selected battery of psychometric tests to measure neuropsychological functioning. OUTCOME AND RESULTS: The exposed group had significantly impaired short term verbal and spatial memory, impaired sustained and divided attention, and impaired motor speed. They also suffered from elevated clinical levels of psychiatric symptomatology, including anxiety, depression and phobic avoidance, and neurological symptoms of tremor, weakness in the limbs, and excessive sweating. CONCLUSIONS: The exposed group suffered from varying degrees of permanent neuropsychological disability, which adversely affects their ability to work and be financially independent. Psychometric measures for monitoring cognitive symptoms are discussed. PMID- 11030455 TI - Electrophysiological indices of persistent post-concussion symptoms. AB - Mild head injuries (MHI) including concussion were once considered transient alterations of function that resulted in no long-term structural or functional effects. This opinion has changed somewhat in recent years, based on the scientific evidence and popular cases in the media that suggest MHI can result in damage that can, in some cases, lead to long-term cognitive sequelae. PURPOSE AND METHODS: An EP/ERP assessment battery is presented as a method for detecting changes in brain function that form the organic basis of persistent post concussion symptoms (PCS). The primary focus of the paper was directed towards using this battery to determine whether or not brain function in individuals with persistent PCS was different than responses of individuals that comprised a nomative database. RESULTS: Visual and auditory ERPs and visual EPs were beyond a 2.5 standard deviation normal limit in young and older groups of individuals with persistent post-concussion symptoms. CONCLUSIONS: Evidence of changes in brain function in individuals with persistent post-concussion symptoms is consistent with the position that the post-concussion syndrome has a substantial biological, as opposed to a psychological, basis. PMID- 11030456 TI - Visual disorders after traumatic brain injury in developmental age. AB - Visual disorders secondary to TBI are common, often multiple, associated with complex clinical pictures; in developmental age they may particularly interfere with the development process, and compromise the rehabilitation and outcome processes. The aim of this study is to identify visual disorders in 56 post traumatic children admitted to the Traumatic Brain Injury Unit. All patients underwent a complete clinical, neurological and neuroophthalmological assessment. Correlations were studied between visual disorders and clinical parameters of acute phase, age at trauma, neuroimaging data and outcome, in order to identify possible risk factors for their occurrence. A high incidence was found of complex visual disorders following head trauma: decreased visual acuity, together with a convergence oculo-motor deficit, is the most frequently detected deficit. Trauma severity, indicated by duration of coma, is associated with most of the visual disorders and appears to be the main risk factor conducive to their appearance. PMID- 11030457 TI - Phenytoin-induced asterixis--uncommon or under-diagnozed? AB - Asterixis is a recognized but uncommon clinical sign of phenytoin toxicity. A case is presented in which asterixis and acute cerebellar dysfunction occurred when the phenytoin levels reached the toxic range. It disappeared when the drug levels normalized. Asterixis is now classified as a form of negative myoclonus, which is characterized by irregular myoclonic lapses of posture. The neurochemistry and physiology of phenytoin causing asterixis has yet to be elucidated. PMID- 11030458 TI - Platelet annexin V: the ins and outs. AB - One of the most important signals accompanying platelet activation is the increase in intracellular calcium, and it is known that calcium-binding proteins play an important role in linking this calcium signal to the final platelet responses. Annexins are highly conserved calcium-binding proteins, of which annexin V is the major annexin in human platelets. The last few years have seen an accumulation of information on and speculations about the roles of annexins in both intracellular and extracellular locations. Additionally, annexin V has proven very useful as a marker for apoptosis and platelet activation. It also has potential for the delivery of thrombolytic agents to thrombi and for imaging thrombi as they form in the human body. This review distinguishes between the potential intracellular role of annexin V in platelet activation, and the uses of annexin V in particular to detect apoptosis. PMID- 11030459 TI - Update on collagen receptor interactions in platelets: is the two-state model still valid? AB - This review summarises some of the key developments that have taken place in our understanding of platelet-collagen interactions within the last 18 months. Within this time, the major activatory collagen receptor glycoprotein VI (GPVI) has been sequenced and shown to reconstitute collagen responses in a megakaryocytic cell line. It is a member of the Ig superfamily of proteins, with two extracellular Ig domains, and is constitutively associated with the Fc receptor gamma-chain (FcR gamma-chain). GPVI signals through a pathway that shares many features with those of immune receptors, with critical roles for Syk and the adapters LAT and SLP-76 in the activation of PLCgamma2. Significant developments have also taken place in regard to the role of the major adhesion receptor for collagen, the integrin alpha2beta1 (also known as GPIa-IIa). An alpha2beta1-selective collagen-based peptide has been developed and co-crystallised with the I-domain of the alpha2 subunit. Polymorphisms in alpha2 have been shown to cause wide variation in expression of alpha2beta1, with the alpha2 allele T807/A873 leading to a high level of the integrin and increased risk of stroke in young people. Activation of platelets by a wide range of agonists has been shown to increase the affinity of alpha2beta1 to intermediate or high affinity states. This has important implications for the two-site, two-state model of collagen-platelet interactions. A new model is proposed in which collagen binds initially to either alpha2beta1 or GPVI, leading to subsequent binding to the other receptor and conversion of the integrin to a high affinity state. In this model, both receptors generate intracellular signals which support platelet activation. PMID- 11030460 TI - Characterization and quantitation by flow cytometry of membranous microparticles formed during activation of platelet suspensions with ionophore or thrombin. AB - The typical data presentation by flow cytometry of platelet suspensions stimulated with calcium ionophore A-23187, thrombin, C5b-9, or other agonists shows a unimodal decrease (or 'left-shift') in forward angle light scatter. Many reports in the literature interpret these findings as indicative of the appearance of small membranous microparticles generated from the platelets as part of the activation response. Investigators may attempt to quantify the microparticles by calculation of the percentage of counts falling below a gate set around the forward angle light scatter distribution of fresh, non-activated platelets. We believe that this approach can lead to erroneous results unless the total particle count in the sample is also determined. The true change in total particle count in a platelet sample is easily estimated on the flow cytometer by adding a known amount of fluorescent beads to the platelet suspension and noting a change in the ratio of bead versus non-bead event counts as a result of the stimulus added. Using this technique under settings considered routine for platelet analysis on a FACScan flow cytometer (Becton-Dickinson), we have found that particle counts increased very little (less than a doubling) in platelet suspensions stimulated with 1-10 microM A-23187 or 0.01-0.5 U/ml thrombin or with sera from patients with diagnosed heparin-induced thrombocytopenia (HIT). Only by employing high sensitivity settings for signal thresholding on orthogonal light scatter, combined with fluorescence gating on high prevalence surface antigens, were we able to detect significant increases (5- or 6-fold) in total particle count in the same experiments. The new events we observed were separated by a decade or more (log scale) from intact platelets on the light scatter plots and fluorescence histograms in a bimodal distribution. We postulate that the unimodal shifted population of events seen under routine settings after stimulation with ionophore is really degranulated platelets, and only the much smaller new modal subpopulation represents microparticles released as new entities into the platelet suspension. We conclude that without high sensitivity settings for data acquisition, it is most likely incorrect to claim that the left-shifted events on flow cytometry light scatter plots appearing contiguous with the distribution of activated platelets are released microparticles. PMID- 11030461 TI - Effect of endothelin-1 on human platelet shape change: reversal of activation by naftidrofuryl. AB - Naftidrofuryl (Praxilene; NAF) significantly improves claudication distance in patients with peripheral vascular disease (PVD). Endothelin-1 (ET-1) is a powerful endogenous vasoconstrictor and the circulating levels of ET-1 are elevated in patients with vascular disease. Platelet rich plasma (PRP) was prepared from healthy volunteers. NAF at concentrations similar to therapeutic levels (3.5-14 micromol/l), inhibited (P < 0.02) platelet activation (as indicated by a fall in median platelet volume, MPV) induced by ET-1 (0.4 micromol/l) alone. NAF also inhibited (P <0.0001) shape change (PSC; an early phase of platelet activation, characterised by an increase in MPV) induced by ET 1 (0.4 micromol/l) in combination with ADP (0.05-0.15 micromol/l) or serotonin (0.03-0.13 micromol/ l). We assessed the effect of ET(A) (BQ123, 50 nmol/l) or ET(B) (BQ788, 50 nmol/l) receptor antagonists on PSC induced by ET-1 alone. Both antagonists significantly inhibited PSC. We conclude that ET-1 activates human platelets. Both ET(A) and ET(B) receptors probably contribute to this response by a complex mechanism that requires further elucidation. NAF antagonises the action of ET-1 on human platelets. These actions may contribute to the beneficial effects of NAF in PVD. PMID- 11030462 TI - Platelet-dependent modulation of polymorphonuclear leukocyte chemiluminescence. AB - Human blood platelets decreased luminol-enhanced chemiluminescence of human polymorphonuclear leukocytes (PMNL) stimulated with FMLP or Ca2+-ionophore A23187 by 56 or 47%, respectively. Horseradish peroxidase potentiated the decreasing effect of platelets on A23187-stimulated PMNL (92% inhibition) or reversed inhibition of FMLP-induced chemiluminescence to 94% potentiation, indicating dependence of platelet activity on availability of extracellular peroxidase. Moreover, platelet activity may depend also on the extent of platelet activation, as non-activated platelets (in the presence of FMLP) were found to potentiate PMNL-generated chemiluminescence, while platelets activated with A23187 displayed the opposite effect. Interference of platelets with formation and liberation of superoxide anion was indicated by platelet-modified isoluminol chemiluminescence. Superoxide dismutase with catalase and sodium azide were used, respectively, to differentiate the intracellular and the extracellular part of the chemiluminescence signal. Platelets were found to be capable of modifying both components of chemiluminescence, i.e., oxygen metabolites produced on the plasma membrane as well as on membranes of intracellular granules. PMID- 11030463 TI - ADP-induced platelet shape change: an investigation of the signalling pathways involved and their dependence on the method of platelet preparation. AB - Shape change is an important early event in platelet activation. In this study we show that the Ca2+ chelator BAPTA and the Rho-kinase inhibitor Y-27632 inhibit ADP-induced myosin light chain (MLC) phosphorylation and platelet shape change through distinct pathways and with distinct kinetics. Ca2+ is largely responsible for the initial onset of shape change, whilst Rho-kinase plays a major role in the maintenance of the response. The relative contribution of these two pathways to each stage of the response was dependent on the method of platelet preparation, but in all cases shape change was shown to be downstream of the P2Y1 receptor. Similar observations were made in murine platelets. The shape change response was modulated via changes in cAMP levels, possibly via the P2TAC receptor, but not by tyrosine phosphorylation. We conclude that ADP-induced shape change occurs via the P2Y1 receptor, which can be differentially coupled to Rho kinase and Ca2+-linked pathways dependent on the method of platelet preparation. PMID- 11030465 TI - Potential of substance P antagonists as antiemetics. AB - The introduction of serotonin 5-HT3 receptor antagonists into clinical practice allowed for a dramatic improvement in the management of nausea and vomiting. Despite this, postoperative and chemotherapy-induced emesis remains a significant, unresolved issue in many patients even when a combination of antiemetic drugs is used. Numerous neurotransmitters have been implicated in triggering emesis; however, the tachykinin substance P, by virtue of its localisation within both the gastrointestinal vagal afferent nerve fibres and brainstem emetic circuitry, and its ability to induce vomiting when administered intravenously, is thought to play a key role in emetic responses. Because substance P is the most likely endogenous ligand for the neurokinin-1 (NK1) receptor, the development of nonpeptide NK1 receptor antagonists led scientists to evaluate these compounds as antiemetics. The five NK1 receptor inhibitors that have been studied initially in humans are: vofopitant (GR-205171), CP-122721, ezlopitant (CJ-11974), MK-869 (L-754030) and its prodrug L-758298. Except for monotherapy in acute cisplatin-induced emesis, this new class of drugs has proven to be highly effective in the control of both chemotherapy-induced nausea and vomiting, and postoperative nausea and vomiting. No major adverse event was reported in the preliminary trials. Further investigation is mandatory in order to assess the optimal treatment regimen and to make sure the wide spectrum activity of the NK1 receptor inhibitors does not cause significant adverse effects in the context of the treatment of nausea and vomiting. PMID- 11030464 TI - Combination chemotherapy for hepatitis B virus: the path forward? AB - Hepatitis B virus (HBV) was identified as a cause of viral hepatitis more than 30 years ago and hepatitis B vaccines have been available for almost 20 years, but HBV infection continues to be a global health problem, responsible for about 1.2 million deaths annually. By the end of this year, almost 400 million people- about 5% of the world's population and more than ten times the number infected with human immunodeficiency virus (HIV)--will be infected with HBV. Chemotherapy remains the only treatment option for controlling chronic HBV infection once acquired, but none of the many different chemotherapeutic strategies used in the past has proven consistently successful. Prospects for successful treatment of HBV have improved dramatically during the past decade due to the development of new, well tolerated and efficacious anti-HBV drugs, and to advances in our understanding of HBV replication and pathogenesis. The newer anti-HBV drugs are capable of reducing viral loads very rapidly, but the initial response is invariably followed by very much slower elimination of residual virus. As more effective anti-HBV drugs become available, the emergence of drug resistance during the slower phase of HBV elimination will probably become the most significant obstacle in the way of eventual control of HBV infection. Experience with HIV indicates that combination chemotherapy may suppress or eliminate drug resistance and methods for pre-clinical and clinical assessment of anti-HBV drug combinations are being developed. Basic research into mechanisms of drug action and interaction should assist in the design and optimisation of combination chemotherapy for HBV infection, for which additional new anti-HBV drugs will undoubtedly be required in future. PMID- 11030466 TI - Immune tolerance therapy for haemophilia. AB - The development of anti-factor VIII and anti-factor IX allo-antibodies in haemophilia A and B, respectively, remains a serious complication of treatment for these two X-linked haemostatic disorders, with major clinical and economic consequences. Treatment of this potentially fatal complication remains one of the greatest challenges facing haematologists at the beginning of the 21st century. Immune tolerance induction (ITI) therapy has been generally accepted as the best available treatment, extinguishing the inhibitor and permitting a resumption of standard dosing schedules. Although there have been several established protocols for ITI therapy developed over the last quarter century, the optimal scheme in terms of safety, clinical efficacy and pharmacoeconomic considerations has yet to be determined. PMID- 11030467 TI - Oral versus topical NSAIDs in rheumatic diseases: a comparison. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most commonly prescribed drugs worldwide and are responsible for approximately one-quarter of all adverse drug reaction reports. NSAIDs are widely prescribed for patients with rheumatic disease--a population at increased risk for serious gastrointestinal (GI) complications. Topical administration of NSAIDs offers the advantage of local, enhanced drug delivery to affected tissues with a reduced incidence of systemic adverse effects, such as peptic ulcer disease and GI haemorrhage. NSAIDs administered topically penetrate slowly and in small quantities into the systemic circulation; bioavailability and maximal plasma NSAID concentration after topical application are generally less than 5 and 15%, respectively, compared with equivalent oral administration. Product formulation may have a dramatic impact, not only on absorption rates but also on penetration depth. Compared with oral administration, topical application leads to relatively high NSAID concentrations in the dermis. Concentrations achieved in the muscle tissue below the site of application are variable, but are at least equivalent to that obtained with oral administration. NSAIDs applied topically do reach the synovial fluid, but the extent and mechanism (topical penetration versus distribution via the systemic circulation) remain to be determined. In addition, marked interindividual variability was noted in all studies; percutaneous absorption may be strongly influenced by individual skin properties. In general, interpretation of clinical studies measuring efficacy of topical NSAIDs in rheumatic disease states is difficult because of a remarkably high placebo response rate, use of rescue paracetamol (acetaminophen), and significant variability in percutaneous absorption and response rates between patients. Overall efficacy rates attributable to topical NSAIDs in patients with rheumatic disorders ranged from 18 to 92% of treated patients. Topically applied NSAIDs have a superior safety profile to oral formulations. Adverse effects secondary to topical NSAID application occur in approximately 10 to 15% of patients and are primarily cutaneous in nature (rash and pruritus at site of application). GI adverse drug reactions are rare with topically applied NSAIDs, compared with a 15% incidence reported for oral NSAIDs. Available clinical studies suggest, but do not document, equivalent efficacy of topical over oral NSAIDs in rheumatic diseases. PMID- 11030469 TI - Treatment of Chlamydia trachomatis infections in pregnant women. AB - The intent of this article is to provide an overview of the epidemiology and pharmacotherapy, including cost analyses, of Chlamydia trachomatis infections in pregnant women. Chlamydia is a common sexually transmitted infection. For pregnant women, there are concerns both for the mother (post-partum endometritis, horizontal transmission) and the newborn (conjunctivitis, delayed pneumonia). Therapeutic options are restricted because of the fetus and include multi-day treatment with erythromycin, amoxicillin, clindamycin or single dose azithromycin. Clinical cure rates with these options are 86, 92, 93 and 95%, respectively. Pharmacoeconomic analyses have been conducted to determine if the initial increase in acquisition cost of azithromycin (approximately 3-fold higher than erythromycin or amoxicillin) is offset by improvement in compliance and drug efficacy. Clindamycin has received little attention because of its expense (4 fold more than azithromycin). Analyses have been retrospective. As models incorporate more complications of failure to cure, azithromycin increasingly becomes more cost effective and is our recommended treatment. PMID- 11030468 TI - Proficient and cost-effective approaches for the prevention and treatment of venous thrombosis and thromboembolism. AB - Thrombosis is clearly a common cause of death in the US. It is obviously of major importance to define the aetiology of deep vein thrombosis (DVT) as (i) many of these events are preventable if appropriate therapy, dependent upon the risk factors known is utilised; (ii) appropriate antithrombotic therapy will decrease risks of recurrence; (iii) the type of defect(s) and risk(s) will determine length of time the patient should remain on therapy for secondary prevention and (iv) if the defect is hereditary appropriate family members can be assessed. Aside from mortality, significant additional morbidity occurs from DVT including, but not limited to, stasis ulcers and other sequelae of post-phlebitic syndrome. Numerous studies have provided evidence that medical patients and patients undergoing surgery or trauma are at significant risk for developing DVT, including pulmonary embolism (PE). Thus, an important task for the clinician is to prevent DVT and its complications. It is important to define risk groups where prophylaxis must be considered. The attitudes and beliefs towards prophylaxis show great regional variations. This is true for the definition of risk groups, the proportion of patients receiving prophylaxis and prophylactic modalities used. For this reason, various 'consensus conference' groups have attempted to alleviate these problems; the primary mission of consensus guidelines is to provide optimal direction to the clinician in the setting of clinical practice. If the practice guidelines generated are successful they will assist clinicians in decision-making for their patients, and they will also provide protection against unjustified malpractice actions. Therapy may be complex, as clinical studies continue to identify more effective treatments. This review includes currently accepted approaches to the treatment of DVT. The clinical course of DVT is highly dynamic. When the response to therapy is not as expected, more than one cause of DVT may be present in a patient. Treatment must address the primary coagulopathy as well as any precipitating factors. The risk of pharmacological intervention must be balanced against potential benefit. If the incidence of DVT in a given disorder is low and if the mortality rate is similarly low, therapy with an agent known to be associated with a high risk for complications, such as warfarin, would not be indicated. If DVT is seen primarily after surgery or in other high-risk situations, therapy might be limited to a fixed time period. However, if the ongoing risk of DVT remains high or if a history of recurrent DVT dictates, lifelong therapy might be indicated. The recommendations presented are based upon published controlled trials; however, indications for therapy and therapeutic agents of choice will continually evolve. By applying the principles outlined in this review, substantial cost savings, reduction in morbidity and reductions in mortality should occur. PMID- 11030470 TI - Nateglinide. AB - Nateglinide is a novel D-phenylalanine derivative that inhibits ATP-sensitive K+ channels in pancreatic beta-cells in the presence of glucose and thereby stimulates the prandial release of insulin. Nateglinide reduces fasting and mealtime blood glucose levels in animals, healthy volunteers, and patients with type 2 (non-insulin-dependent) diabetes mellitus, and produces prompt prandial insulin responses with return to baseline insulin levels between meals. In randomised, double-blind 24-week studies in patients with type 2 diabetes, oral nateglinide 120 mg 3 times daily before meals improved glycaemic control significantly relative to placebo. Nateglinide 120 mg plus metformin 500 mg, both 3 times daily, conferred greater glycaemic improvement than either drug given alone, and nateglinide 60 or 120 mg 3 times daily plus metformin 1 g twice daily was superior to metformin plus placebo. Nateglinide 120 mg 3 times daily significantly reduced hyperglycaemia relative to placebo in a 16-week double blind study in patients with type 2 diabetes mellitus. Combination therapy with troglitazone 600 mg daily produced significantly better glycaemic control than either drug given as monotherapy. Mild hypoglycaemia was the most frequently reported adverse event (1.3% of patients) after treatment with nateglinide 120 mg 3 times daily in a 16-week clinical study. No clinically significant abnormalities in laboratory results, ECGs, vital signs or physical examination findings have been noted in patients taking the drug. PMID- 11030471 TI - Meropenem: an updated review of its use in the management of intra-abdominal infections. AB - Meropenem is a carbapenem antibacterial agent with a broad spectrum of activity which encompasses gram-negative, gram-positive and anaerobic bacteria. Like other carbapenems, meropenem is stable against chromosomal and extended-spectrum beta lactamases. In patients with moderate to severe intra-abdominal infections, empirical monotherapy with meropenem achieved clinical response rates ranging from 91 to 100% in 7 randomised comparative trials. Efficacy rates were similar to those of imipenem/cilastatin (94 to 97%), clindamycin plus tobramycin (93%) and, overall, to cefotaxime plus metronidazole (75 to 100%), although there were differences between trials versus this combination regimen. According to limited data, meropenem also achieved clinical response rates of over 80% in patients with severe intra-abdominal infections. Meropenem is well tolerated, the most common adverse events being diarrhoea, rash, nausea/vomiting and inflammation at the injection site which are reported in <2.5% of patients each. Meropenem also has an improved CNS tolerability profile compared with imipenem/cilastatin. CONCLUSIONS: Extensive comparative clinical data demonstrate that meropenem can be used effectively as empirical monotherapy in moderate to severe intra abdominal infections. It also shows potential in the most severe forms of infection, although experience in this infection type remains limited. Compared with standard combination regimens, meropenem offers the benefits of ease of administration without the need for monitoring. It also offers improved CNS tolerability compared with imipenem/cilastatin with the option of a higher maximum dosage, which may be a particular advantage in patients with severe intra abdominal infections. PMID- 11030472 TI - Metoprolol: a review of its use in chronic heart failure. AB - Metoprolol, a relatively selective beta1-blocker, is devoid of intrinsic sympathomimetic activity and possesses weak membrane stabilising activity. The drug has an established role in the management of essential hypertension and angina pectoris, and more recently, in patients with chronic heart failure. The effects of metoprolol controlled-release/extended-release (CR/XL) in patients with stable, predominantly mild to moderate (NYHA functional class II to III) chronic heart failure have been evaluated in the large Metoprolol CR/XL Randomised Intervention Trial in Congestive Heart Failure (MERIT-HF) trial and the much smaller Randomized Evaluation of Strategies for Left Ventricular Dysfunction (RESOLVD) pilot study. Treatment with metoprolol CR/XL was initiated at a low dosage of 12.5 to 25 mg once daily and gradually increased at 2-weekly intervals until the target dosage (200 mg once daily) or maximal tolerated dosage had been attained in patients receiving standard therapy for heart failure. At 12 months, metoprolol CR/XL was associated with a 34% reduction in relative risk of all-cause mortality in patients with chronic heart failure due to ischaemic or dilated cardiomyopathy in the MERIT-HF trial. The incidence of sudden death and death due to progressive heart failure were both significantly decreased with metoprolol CR/XL. Similarly, a trend towards decreased mortality in the metoprolol CR/XL group compared with placebo was observed in the RESOLVD trial. Data from small numbers of patients with severe (NYHA functional class IV) heart failure indicate that metoprolol CR/XL is effective in this subset of patients. However, no firm conclusions can yet be drawn. Improvement from baseline values in NYHA functional class, exercise capacity and some measures of quality of life with metoprolol CR/XL or immediate-release metoprolol were significantly greater than those with placebo. The drug is well tolerated when treatment is initiated in low dosages and gradually increased at intervals of 1 to 2 weeks. CONCLUSIONS: Metoprolol CR/XL effectively decreases mortality and improves clinical status in patients with stable mild to moderate (NYHA functional class II or III) chronic heart failure due to left ventricular systolic dysfunction, and the drug is effective in patients with ischaemic or dilated cardiomyopathy. Although limited data indicate that metoprolol CR/XL is effective in patients with severe (NYHA functional class IV) chronic heart failure, more data are needed to confirm these findings. Treatment with metoprolol CR/XL significantly reduced the incidence of sudden death and death due to progressive heart failure. PMID- 11030474 TI - Nutritional composition and micronutrient status of home made and commercial weaning foods consumed in Tanzania. AB - About 50% of young children in Tanzania suffer from protein-energy undernutrition (PEU) while more than 45% of children under the age of five suffer from various micronutrient deficiency disorders. The immediate cause of these conditions is inadequate intake and poor utilization of nutrients, which begins in the weaning period and amplifies in the subsequent years. This study was conducted to assess the potential of some home made and commercial weaning foods commonly consumed in Tanzania to supply adequate amounts of both macro- and micronutrients as recommended in the Tanzania and FAO/WHO Codex Alimentarius Standards for cereal/milk-based weaning foods. Six types of home made weaning foods, maize, cassava, millet, sorghum and millet-sardine-peanut composite gruels and plantain pap, and four types of commercial weaning foods, Cerelac- 1, Cerelac-2, Lactogen 1 and Lactogen-2, popularly consumed in Tanzania, were chemically assayed for proximate composition, energy and mineral density. Results of the study indicated that, both the home made and commercial weaning foods were good sources of macro- and micronutrients. When compared with the Codex Alimentarius and Tanzania Bureau of Standards specifications for weaning foods, both home made and commercial weaning foods had some shortcomings in terms of nutrient composition and energy balance. Many of the foods were low in fat. Fe, Ca, Zn and P but high in crude fiber, carbohydrate and magnesium. Ca, Fe and Zn were the most common deficient macro/micronutrients in the home made weaning foods. In spite of these shortcomings, most of the home made and commercial weaning foods were nutritionally sound since they could provide reasonable percentages of the recommended daily allowances for macro/micronutrients and energy. It is suggested that, more efforts must be directed towards increasing the concentration of Ca, Fe and Zn in the home made weaning foods through supplementation of the starchy staples with mineral rich foods. Meanwhile, the parents, caretakers and health workers should be educated on the selection and preparation of nutritious, balanced weaning foods and good weaning practices. PMID- 11030473 TI - Desirudin: a review of its use in the management of thrombotic disorders. AB - Desirudin, a recombinant hirudin used in the prevention and management of thromboembolic disease, is a thrombin inhibitor which binds directly and with high affinity to clot-bound and fluid phase thrombin. As a prophylaxis in patients undergoing hip replacement surgery, desirudin was significantly more effective in reducing the incidence of deep vein thrombosis (DVT) than either unfractionated or low molecular weight heparin. However, results in patients with acute coronary syndromes are less conclusive. A significant reduction with desirudin compared with heparin in the incidence of death or non-fatal (re)infarction at 24 hours in patients with acute myocardial infarction (MI) was reported in the GUSTO (Global Use of Strategies to Open Occluded Coronary Arteries) IIb trial but not in the TIMI (Thrombolysis and Thrombin Inhibition in Myocardial Infarction) 9B trial. Despite the early reduction shown in GUSTO IIb, desirudin was not associated with an improved long term clinical benefit at 30 days compared with heparin. Similar results were seen in patients with unstable angina/non-Q-wave MI enrolled in the GUSTO IIb trial. In addition, desirudin and heparin showed similar efficacy in preventing restenosis 30 weeks after coronary angioplasty for unstable angina, despite desirudin being associated with a significant reduction in the rate of cardiac events within the first 96 hours. Desirudin is as well tolerated as heparin with a similar incidence of moderate and severe bleeding, intracranial haemorrhage or stroke reported when trialled in the prevention of DVT associated with hip replacement surgery or the treatment of acute coronary syndromes. However, in the GUSTO IIb trial a significantly higher incidence of transfusions was observed in patients with unstable angina/non-Q wave MI. CONCLUSIONS: Desirudin is clearly more effective than heparin in the prevention of DVT in patients undergoing elective hip replacement, although cost factors may influence its ultimate place in therapy. In the treatment of acute coronary syndromes the role of desirudin is less certain; however, it may be useful for patients in whom heparin therapy is not a viable option. PMID- 11030475 TI - Dietary fiber content of commonly fresh and cooked vegetables consumed in India. AB - Legumes, leafy vegetables, roots and tubers, gourds and other vegetables were analyzed for total (TDF), soluble (SDF) and insoluble (IDF) dietary fiber contents, both before and after cooking either by a conventional open-pan method or by pressure cooker. Data revealed a significant increase in SDF fraction with a concomitant decrease in the IDF fraction upon cooking by both the methods employed. Although the decrease in IDF matched the increase in SDF values in some cases, it was found to be more in vegetables categorized as 'other'. The dietary fiber values have also been reported on a fresh weight basis which may serve as a guideline for calculating dietary intake of each component by the consumer. PMID- 11030476 TI - Optimization of the solid state fermentation process to obtain tempeh from hardened chickpeas (Cicer arietinum L.). AB - Solid state fermentation (SSF) represents a technological alternative for a great variety of cereals and legumes, or combination of them, to improve their nutritional quality and to obtain edible products with palatable sensorial characteristics. The objective of this work was to find the best conditions of fermentation temperature and time to obtain tempeh from hardened chickpeas (Cicer arietinum L.) applying SSF. Response surface methodology (RSM) was applied over three response variables (phytic acid, in vitro protein digestibility and available lysine) to find best conditions of fermentation to carry out the process. A central composite experimental design with two factors [X1 = temperature (31-36 degrees C) and X2 = time (48-72 h)] in five levels (2 factorials, 2 axial, I central) was used. Spores from Rhizopus stolonifer were suspended in distilled water (1 x 10(6) spores/mL) and used as starter. According to regression models, minimum and maximum levels of the response variables were 1.24-2.66 mg phytic acid/g of sample DM, 77.6-83.5% in vitro protein digestibility and 2.18-4.63 g available lysine/16 g N. The superposition of contour plots of each one of the response variables allowed researchers to find, graphically, the best conditions for the SSF process: 35.8 degrees C for 42.7 h. PMID- 11030477 TI - Hard-to-cook phenomenon in chickpeas (Cicer arietinum L): effect of accelerated storage on quality. AB - Storage, at high temperature (> or = 25 degrees C) and high relative humidity (> or = 65%), causes development of hard to cook (HTC) phenomenon in grain legumes. The objective of this work was to study the effect of storage simulating tropical conditions on chickpeas quality. The hardening of the Surutato 77, Mocorito 88, and Blanco Sinaloa 92 chickpea varieties was produced using adverse storage (32 +/- 1 degrees C, RH = 75%, 160 days) conditions. For all samples, the Hunter 'L' values decreased and deltaE values increased during storage, meaning a loss of color lightness and development of darkening. Accelerated storage caused a significant decrease in the water absorption capacities and cooking times of whole seeds, cotyledons and seed coats of all samples, being more pronounced in The Blanco Sinaloa 92 variety. Furthermore, storage produced significant decreases in the seed coat tannin content of the three materials; this parameter increased significantly in the cotyledon. In all samples, the levels of phytic acid decreased significantly with the seed hardness. Hardening of chickpea grains caused a decrease in the in vitro protein digestibilities of all varieties. These results suggest that both the cotyledon and seed coat play a significant role in the process of chickpea hardening. Blanco Sinaloa 92 and Mocorito 88 might be classified as varieties with high and low proneness, respectively, to the development of the HTC condition. PMID- 11030478 TI - Hemagglutinins in vegetative tissues of Bengal gram. AB - Hemagglutinating activity was identified in the roots and vegetative tissues of Bengal gram using rabbit erythrocytes. The activity in the roots appears to be similar to that of the seeds in respect to their sugar inhibition property. Seed and root extracts were found to have fetuin inhibitory agglutinating activity, whereas cotyledons showed different sugar specificity. However, 7-day cotyledons and roots from 3 and 7-day plants were also found to have galactose and N-acetyl D-galactosamine inhibitory activity. Protein profile in different tissues during their developmental stages was analyzed by SDS-PAGE. A major band at 66 kDa was common to all tissues at different stages of their development except 3-day cotyledons where 2 major bands at 22 kDa and 74 kDa were observed. Two prominent bands at 74 kDa and 86 kDa were found along with 60 kDa protein in 3-day old roots whereas a major band at 33 kDa was found in 15-day old roots. Total protein content and specific activity of hemagglutinin were measured. Increase in specific activity with lower protein content was observed in cotyledons of 15-day old plant. Specific activity was eight times higher in the roots and four times more in leaf tissues of 7-day old plants in comparison to 15-day old plants. Stems also showed a significant increase of specific activity (p = 0.013) in 7 day old plants without any significant alteration in the protein content. PMID- 11030479 TI - Chemical evaluation of multimixes formulated from some local staples for use as complementary foods in Nigeria. AB - The nutrient compositions of eight multimixes formulated for use as complementary foods from processed soybeans, cowpeas, maize, sorghum, yams, cocoyams, plantains and sweet potatoes were examined. The foods were processed by sprouting, cooking and fermentation. The samples were separately dried and milled to fine flours. A ratio of 65% cereal, 30% legume and 5% starchy staple (65:30:5) calculated on a protein basis was used to formulate the multimixes. The blends were chemically analyzed using standard procedures. The mixtures containing soybeans had higher protein, lipid, energy, crude fiber and calcium levels but lower carbohydrate content than those mixtures without soybean. The protein and energy levels of the multimixes were higher than those of some commercial infant foods (Cerelac, Farex and Nestrum) in Nigeria but were comparable to that of 'soy-ogi'. The multimixes contained fair quantities of calcium and phosphorus and an adequate amount of some of the essential amino acids. Methionine was the most limiting amino acid in all the mixtures. Blending cowpeas with maize produced a higher protein score than blending cowpeas or soybeans with sorghum. The cowpea/maize/sweet potato mixture had the highest protein score. PMID- 11030480 TI - Nutritional quality of sunflower seed protein fraction extracted with isopropanol. AB - This study investigated the nutritional effect of sunflower seed protein fraction (SSPF) extracted with isopropanol on growth, plasma and tissue lipid profile, protein content and erythrocyte membrane lipid profile of rats. Dehulled sunflower seeds were extracted with isopropanol at 50 +/- 1 degree C resulting in a protein fraction (71.5%) with low residual chlorogenic acid (0.07%) and fiber (3.3%) contents. Rats fed the sunflower seed protein fraction had a similar body weight gain and food efficiency ratios in comparison to those fed casein. Rats fed SSPF in contrast had a significantly higher growth and food efficiency ratio than the rats fed sunflower meal (SM), extracted with hexane. However, dietary proteins exerted a separate effect on plasma total cholesterol, low density lipoprotein (LDL)-cholesterol, low density lipoprotein to high density lipoprotein cholesterol (LDL-C/HDL-C) ratio and triglyceride content. Sunflower seed protein fraction resulted in a significant decrease in plasma cholesterol (p < 0.05) and LDL-cholesterol (p < 0.02) levels compared to the casein fed rats. Membrane phospholipid profile also showed a marked variation with the type of dietary protein. Rats fed SSPF and SM did not show much variation in plasma lipids, plasma proteins, liver and brain lipids and membrane phospholipid concentrations. Protein content, liver and brain lipid profile of the groups fed SSPF and casein were comparable, suggesting that the nutritional value of SSPF is better than SM and equivalent to that of casein. PMID- 11030481 TI - Paying attention to your acetylcholine, Part 2: the function of nicotinic receptors. PMID- 11030482 TI - The implications of genetics studies of major mood disorders for clinical practice. AB - BACKGROUND: This article is a selective review and synthesis of relevant research findings from genetic studies of major mood disorders and the application of these to clinical practice. METHOD: The article discusses the application of genetic research findings in major mood disorders, including epidemiologic and family study risk estimates, risk modifiers, and the concepts of etiologic and phenotypic heterogeneity, to 3 clinical domains: risk counseling, diagnosis, and treatment prediction. RESULTS: Epidemiologic and family studies have provided general risk estimates useful in counseling mood-disordered patients and their relatives. A complete and accurate family pedigree provides more individualized risk estimates for specific cases and is useful in identifying the phenotypic spectrum of the disorder being transmitted in the family. Both proband course parameters and familial loading for psychiatric illnesses may be relevant for the prediction of treatment response. However, the hypothesis of inherited pharmacologic selectivity remains to be proven. CONCLUSION: Genetic studies of mood disorders have not yet provided conclusive evidence of specific susceptibility genes or their pattern of inheritance. However, they have generated information that is useful to clinical practice. PMID- 11030483 TI - A naturalistic comparison of clozapine, risperidone, and olanzapine in the treatment of bipolar disorder. AB - BACKGROUND: Our purpose was to evaluate the overall efficacy and tolerability of novel antipsychotic medications for patients with bipolar disorder type I. METHOD: A retrospective study of the Massachusetts General Hospital Bipolar Clinic database was carried out to identify 50 consecutive treatment trials in patients with DSM-IV bipolar disorder type I who had received adjunctive treatment with risperidone, olanzapine, or clozapine, along with standard mood stabilizers. The treatment charts of those patients (N = 42) were reviewed for details of adverse effects, tolerability, and efficacy of medication. RESULTS: Overall results indicated equivalent efficacy in novel antipsychotic treatments according to change in Clinical Global Impressions scale score. Levels of extrapyramidal symptoms were similar in all groups and occurred in 12/42 patients (28.6%). Prolactin-related side effects were not observed in any patients. There were no cases of affective switch or worsening of mania. Substantial weight gain of more than 10 lb (4.5 kg) was significantly greater in patients treated with olanzapine. CONCLUSION: These results suggest that the efficacy and tolerability of risperidone, olanzapine, and clozapine are similar in patients with bipolar disorder. One major differentiation factor of these drugs appears to be weight gain, particularly between olanzapine and risperidone. This may, in part, also be related to the need to use mood-stabilizing agents, like lithium or divalproex sodium, which may potentiate the weight-gain effect of novel antipsychotics. PMID- 11030484 TI - A randomized, double-blind placebo-controlled trial of moclobemide in patients with chronic fatigue syndrome. AB - BACKGROUND: Chronic fatigue syndrome is characterized by prolonged and disabling fatigue and a range of neuropsychiatric symptoms including depressed and/or irritable mood. To date, no medical or psychotropic therapies have provided clear symptomatic benefit. METHOD: Ninety patients with chronic fatigue syndrome, diagnosed with our system that approximates CDC criteria, participated in a randomized, placebo-controlled, double-blind trial of 450 to 600 mg/day of moclobemide, a novel reversible inhibitor of monoamine oxidase-A. RESULTS: Fifty one percent (24/47) of patients receiving moclobemide improved compared with 33% (14/43) of patients receiving placebo (odds ratio = 2.16, 95% confidence interval [CI] = 0.9 to 5.1). Drug response was best characterized symptomatically by an increase in the subjective sense of vigor and energy rather than a reduction in depressed mood. The effect of moclobemide on subjective energy was detectable within the first 2 weeks of treatment and increased across the course of the study. The greatest reduction in clinician-rated disability was in patients with concurrent immunologic dysfunction (mean difference in standardized units of improvement = 0.8, 95% CI = 0.03 to 1.6). CONCLUSION: Moclobemide produces some improvement in key symptoms experienced by patients with chronic fatigue syndrome. This effect is not dependent on the presence of concurrent psychological distress and is likely to be shared with other monoamine oxidase inhibitors. PMID- 11030485 TI - Possible induction of mania and hypomania by olanzapine or risperidone: a critical review of reported cases. AB - BACKGROUND: Risperidone and olanzapine are atypical antipsychotics that are now widely used in clinical practice. METHOD: A MEDLINE search (1966-1999) showed that, following the introduction of these agents in recent years, 26 cases of induced hypomanic and manic syndromes have been reported during standard olanzapine (N = 10) or risperidone (N = 16) treatment. RESULTS: A critical analysis of these case reports reveals that the effects on mood were sometimes insufficiently documented and that in about half of them (N = 16) evidence is highly suggestive of a causative role of risperidone or olanzapine in the induction of (hypo)manic symptomatology. CONCLUSION: Despite limitations, the available literature confirms intriguing effects of these 2 antipsychotics on mood. The risk factors as well as the mechanisms of action underlying these effects remain to be clarified. PMID- 11030486 TI - Mirtazapine compared with paroxetine in major depression. AB - BACKGROUND: The aim was to compare the efficacy and tolerability of mirtazapine with those of paroxetine. METHOD: 275 outpatients with a diagnosis of major depressive episode (DSM-IV) and a score > or = 18 on the 17-item Hamilton Rating Scale for Depression (HAM-D-17) were randomly assigned to 6 weeks of treatment with mirtazapine (15-45 mg/day) or paroxetine (20-40 mg/day). Efficacy was assessed by the HAM-D-17, Hamilton Rating Scale for Anxiety (HAM-A), and Clinical Global Impressions scales (Severity and Improvement), and analyses were performed on the intent-to-treat sample (127 mirtazapine-treated patients and 123 paroxetine-treated patients). RESULTS: Mean daily doses were 32.7 mg of mirtazapine and 22.9 mg of paroxetine. Thirty patients in the mirtazapine group and 33 in the paroxetine group dropped out. Both drugs were equally effective in reducing symptoms of depression. At week 1, the mean HAM-D-17 total score was significantly lower in mirtazapine- than paroxetine-treated patients (16.5 vs. 18.8, p = .0032). Similarly, significantly more mirtazapine-treated patients were HAM-D-17 responders (> or = 50% decrease from baseline) at weeks 1 (23.2% vs. 8.9%, p = .002) and 4 (58.3% vs. 44.5%, p = .04). Both treatments were equally effective in reducing anxiety. However, the reduction in mean HAM-A total score was significantly greater with mirtazapine than with paroxetine at week 1 (-5.1 vs. -3.5, p = .0435). Tolerability of both treatments was good, with more nausea, vomiting, tremor, and sweating in the paroxetine group and more weight increase and influenza-like symptoms in the mirtazapine group. CONCLUSION: Mirtazapine and paroxetine were equally effective after 6 weeks of therapy and were both well tolerated. A potentially faster onset of overall therapeutic efficacy of mirtazapine was suggested by significant differences between treatments after 1 week of therapy that were due to slightly larger improvements of several core symptoms of depression as well as distinct prevention of treatment-emergent worsening of anxiety and physical components of depression. PMID- 11030487 TI - Psychostimulant augmentation during treatment with selective serotonin reuptake inhibitors in men with paraphilias and paraphilia-related disorders: a case series. AB - BACKGROUND: We describe an open trial of psychostimulants (primarily methylphenidate sustained release [SR]) added to selective serotonin reuptake inhibitors (SSRIs; primarily fluoxetine) during the course of pharmacologic treatment of men with paraphilias and paraphilia-related disorders (PRDs). METHOD: Twenty-six men with paraphilias (N = 14) or PRDs (N = 12) were assessed for life-time mood disorders and attention-deficit/hyperactivity disorder (ADHD) as defined by DSM-IV. All men were assessed at baseline for total sexual outlet and average time per day associated with paraphilia/PRD sexual behaviors. The indications for the addition of a psychostimulant to a stable dose of SSRI included the retrospective diagnosis of ADHD with persistent adult symptoms despite pharmacotherapy with an SSRI (N = 17); residual paraphilia/PRD fantasies, urges, and activities despite SSRI pharmacotherapy (N = 16); the persistence or presence of residual depressive symptoms despite SSRI pharmacotherapy (N = 6); relapse or loss of SSRI efficacy during the treatment of sexual impulsivity disorders (N = 4); and treatment of SSRI-induced side effects (N = 4). RESULTS: SSRI pharmacotherapy (mean +/- SD duration = 8.8+/-11.1 months) had statistically significant effects in diminishing paraphilia/PRD-related total sexual outlet (p < .001) and average time/day spent in paraphilia/PRD sexual behavior (p < .001). Addition of methylphenidate SR (mean dose = 40 mg/day; mean +/- SD duration = 9.6+/-8.2 months) was associated with additional statistically significant effects on paraphilia/PRD-related total sexual outlet (p = .003) and average time per day (p = .04) in addition to improvement of putative residual ADHD and depressive symptoms. CONCLUSION: Methylphenidate SR can be cautiously and effectively combined with SSRI antidepressants to ameliorate paraphilias and paraphilia-related disorders for the indications listed above. PMID- 11030488 TI - Clinical and psychopharmacologic factors influencing family burden in refractory schizophrenia. The Department of Veterans Affairs Cooperative Study Group on Clozapine in Refractory Schizophrenia. AB - BACKGROUND: This study compares the effect of clozapine and haloperidol and identifies other factors related to family burden as experienced by relatives of patients with refractory schizophrenia (DSM-III-R). METHOD: Of 423 patients participating in a multisite randomized clinical trial, 221 identified a family member who was actively involved in their care and who agreed to complete a standardized measure of family burden at 6 weeks and 3, 6, 9, and 12 months after randomization, simultaneous with comprehensive patient assessments. RESULTS: Patient factors most consistently correlated with greater family burden were symptom severity, days living in the community (i.e., not in the hospital), and frequency of family contact. Among family members, clozapine was associated with significantly (p = .048) greater reduction in feelings of dissatisfaction related to providing support to the patient, but not in objective measures of support, amount of worry the patient engendered, or days of missed employment or household activity. Although clozapine reduces symptoms, thus lowering family burden, it also increases days living in the community, which tends to increase family burden, perhaps canceling out the benefit to families of reduced symptoms. CONCLUSION: Clozapine has a small but significant effect on the experience of families of patients. This is the first study to demonstrate that effective pharmacotherapy may be of some benefit to families as well as to patients. PMID- 11030489 TI - Association between premenstrual syndrome and depression. PMID- 11030490 TI - Is antipsychotic drug--induced weight gain associated with a favorable clinical response? PMID- 11030491 TI - Polypharmacy of 2 atypical antipsychotics. PMID- 11030492 TI - Clarification of anticholinergic effects of quetiapine. PMID- 11030493 TI - Side effect profile of enteric-coated divalproex sodium versus valproic acid. PMID- 11030494 TI - Novel antidepressant strategies to optimize outcome. PMID- 11030496 TI - The WeeFIM instrument: its utility in detecting change in children with developmental disabilities. AB - OBJECTIVE: To examine the utility of the WeeFIM instrument ("WeeFIM") in detecting changes in the functional status of children with disability. DESIGN: Prospective longitudinal design with correlation and responsiveness analysis. SETTING: Three facilities providing services to children with developmental disabilities in western New York State. PARTICIPANTS: Two hundred five children (72 girls, 133 boys) with identified medical disabilities receiving special services were administered the WeeFIM. Subjects ranged in age from 11 to 87 months and came from diverse socioeconomic and ethnic backgrounds. Scores for 174 children were available for 3 administrations performed over a 1-year period. MAIN OUTCOME MEASURES: The responsiveness of the WeeFIM instrument was examined using 5 statistical procedures: Reliability Change Index, Proportional Change Index, effect size, standardized response means, and paired t tests. RESULTS: All 5 indexes of responsiveness indicated statistically significant (p < .05) or reliable changes over time. The transfer subscale of the WeeFIM showed a skewed distribution that affected the results for some responsiveness indexes. The advantage, limitations, and assumptions of the responsiveness indexes are described and graphic examples of change over time are presented to validate the responsiveness of the WeeFIM instrument. CONCLUSION: The WeeFIM instrument showed the ability to document change in functional abilities over a 1-year period in children with chronic disabilities. PMID- 11030495 TI - Psychiatric care of patients with depression and comorbid substance use disorders. AB - BACKGROUND: The goal of this study was to describe the sociodemographic and clinical characteristics and routine psychiatric care of depressed patients with or without substance use disorders (SUDs) and to assess the association between the presence of comorbid SUD and the psychiatric management of patients with depression. METHOD: Each of a sample of 531 psychiatrists participating in the Practice Research Network (PRN) of the American Psychiatric Institute for Research and Education was asked to provide information about 3 randomly chosen patients. Data were collected using a self-administered questionnaire, which generated detailed diagnostic and clinical data on 1228 psychiatric patients. Weighted data were analyzed using the SUDAAN software package. Multivariate logistic regression was used to compare depressed patients with and without SUD. RESULTS: A total of 595 patients (48.4%) were diagnosed with depression (DSM-IV criteria). The prevalence of SUD (excluding nicotine dependence) in this group was 18.1%. The group with SUD had a significantly larger proportion of males, young adults, patients seen in public general hospitals, and non-managed care public plans. No significant group differences were found for primary payer, locus of care, length of treatment, type of current or past treatment, and prescription of medications. Only 2.2% of SUD patients were prescribed with an anti-SUD medication (i.e., disulfiram and naltrexone). CONCLUSION: Concomitant SUDs have little effect on the routine psychiatric care of depressed patients. Efforts should be made to improve the identification and management of depressed patients with SUD. PMID- 11030497 TI - Acquisition versus retrieval deficits in traumatic brain injury: implications for memory rehabilitation. AB - OBJECTIVE: To examine whether impaired memory in persons with traumatic brain injury (TBI) is caused by impaired initial acquisition or compromised retrieval from long-term storage. DESIGN: Prospective matched controlled trial. SETTING: Postacute rehabilitation institute. PARTICIPANTS: Patients with moderate to severe TBI (n = 28) and 21 matched, healthy controls (HCs). MAIN OUTCOME MEASURES: Patients with TBI and HC subjects were equated on initial acquisition on a verbal list-learning task. Recall and recognition performance was then evaluated at 30- and 90-minutes after learning. RESULTS: All HC subjects and 20 subjects with TBI (TBI-MET) were able to meet the learning criterion, but the TBI MET group took significantly more trials than HC subjects to do so. However, after equating groups on acquisition, the TBI-MET group did not differ from controls on recall and recognition at both the 30- and 90-minute delays. Eight TBI subjects showed severe learning deficits (TBI-NOT MET) because they never learned the task, and showed significantly impaired recall and recognition performance. The 2 TBI groups did not differ on measures of severity of injury, but the TBI-NOT MET group performed significantly below the TBI-MET group on executive functioning. Rate of forgetting did not differ across the 3 groups. CONCLUSIONS: Results suggest that memory impairment after TBI is caused primarily by deficiencies in initial acquisition of verbal information rather than in compromised retrieval. The findings have significant implications for the rehabilitation and treatment of individuals with TBI. PMID- 11030498 TI - Sleep apnea syndrome in chronic spinal cord injury: associated factors and treatment. AB - OBJECTIVE: To determine the prevalence of sleep apnea in a sample of persons with chronic spinal cord injury (SCI) of varying injury levels and degrees of impairment. DESIGN: Cross-sectional study. SETTING: Inpatient SCI rehabilitation unit. PARTICIPANTS: Twenty men with SCI (motor complete and incomplete; American Spinal Injury Association classes A-D) of at least 1 year's duration, randomly selected from patients with SCI undergoing elective hospitalization. MAIN OUTCOME MEASURES: Apnea index, determined by sleep study (including chest wall movement, airflow, oxygen saturation), and daytime sleepiness, determined by Epworth sleepiness score. RESULTS: Eight subjects (40%) had sleep apnea, manifested by elevated apnea index (mean +/- SD, 17.1 +/- 6.9) and excessive daytime sleepiness. Sleep apnea was commonly diagnosed in motor-incomplete injuries. A trend (p = .07) existed toward a greater prevalence of sleep apnea with tetraplegia. Age and body mass index were not associated with sleep apnea. CONCLUSION: The prevalence of sleep apnea in men with chronic SCI admitted for nonrespiratory elective hospitalization is high relative to the general population. PMID- 11030499 TI - A randomized trial assessing the effects of 4 weeks of daily stretching on ankle mobility in patients with spinal cord injuries. AB - OBJECTIVE: To determine the effect of 4 weeks of 30 minutes of daily stretching on ankle mobility in patients with recent spinal cord injuries (SCIs). DESIGN: Assessor-blinded randomized controlled trial. SETTING: Two spinal injury units in Sydney, Australia. PATIENTS: Consecutive sample of 14 recently injured patients with paraplegia and quadriplegia. INTERVENTION: Treated ankles were stretched continuously into dorsiflexion with a torque of 7.5 N x m for 30 minutes each weekday for 4 weeks. Contralateral ankles received no stretches. MAIN OUTCOME MEASURES: Passive torque-angle curves for both ankles were obtained at study commencement, then at weeks 2, 4, and 5 (ie, during, at the end of, and 1 week after the stretching program). Torque-angle measurements were obtained with the knee extended and flexed. Mean values for parameters (baseline angle, angle at 10 N x m, slope) describing the characteristics of the torque-angle curves were derived for each knee position. Changes from pretest to each subsequent test were calculated, as well as 95% confidence intervals (CIs) for differences in these changes between stretched and controlled ankles. RESULTS: The stretching intervention did not significantly change any of the 3 parameters describing the torque-angle curves of the ankle in either knee position. At the beginning of the study, the mean (+/-SD) angles obtained with the application of a standardized torque with the knee extended for the control and stretch ankles were 105 degrees (+/- 10.4 degrees) and 106 degrees (+/- 9.8 degrees), respectively. After 4 weeks, these values were 106 degrees (+/- 10.6 degrees) and 107 degrees (+/- 10.6 degrees) (mean difference in change of angle = 0 degrees; 95% CI, -3.3 degrees to 3.3 degrees). CONCLUSION: Thirty minutes of daily stretching for 4 weeks does not significantly change ankle mobility in recently injured patients with SCIs. PMID- 11030500 TI - Cerebral blood volume and oxygenation among poststroke hemiplegic patients: effects of 13 rehabilitation tasks measured by near-infrared spectroscopy. AB - OBJECTIVE: To measure with near-infrared spectroscopy (NIRS) the changes in cerebral hemodynamics and oxygenation in the prefrontal cortex of poststroke patients with hemiplegia performing several rehabilitation tasks and to analyze the different effects of the tasks. DESIGN: Case series pilot study. SETTING: Hospitals and facilities near Tsukuba, Japan. PARTICIPANTS: Twenty-four healthy volunteers and 44 patients with hemiplegia. INTERVENTIONS: For healthy volunteers, conventional rehabilitation tasks of head-up tilt (HUT), calculation, and ergometer. For patients with hemiplegia, these 3 tasks plus reading aloud, listening to music, reciprocal extension, nonparalyzed extension, passive range of motion, pulley, bridge, facilitation, stand-up, and gait. MAIN OUTCOME MEASURES: Changes in cerebral blood volume (CBV) and cerebral oxygen volume (COV) in the prefrontal region sensed by a noninvasive NIRS device placed midforehead in healthy volunteers or on the impaired side in patients with hemiplegia. Computer analysis of the quality and quantity of the CBV and COV change patterns. RESULTS: In healthy subjects, the change patterns of the 3 tasks were clearly different: decrease in COV with HUT, limited increase in CBV and COV with calculation tasks, and gradual increase in CBV and COV with ergometer tasks. In patients with hemiplegia, significant (positive) CBV changes were observed in ergometer, facilitation, stand-up, and gait and significant (negative) changes with Romover. Significant (positive) COV changes were observed in ergometer and facilitation and (negative) in HUT. CONCLUSIONS: NIRS is useful for monitoring the change in regional hemodynamics and oxygenation in rehabilitation; some tasks commonly used in rehabilitation, such as ergometer and facilitation, increase both CBV and COV in the affected prefrontal cortex of patients with hemiplegia. PMID- 11030501 TI - The relation between impairments and functional outcomes poststroke. AB - OBJECTIVE: To assess the influence of initial stroke impairments on the severity of basic and higher level functional deficits over time and to determine the cumulative impact on functional deficits beyond severity of motor deficits alone. DESIGN: Observational study. SETTING: Twelve participating hospitals in the Greater Kansas City area, as part of the Kansas City Stroke Study (October 1995 March 1998). PARTICIPANTS: Individuals (n = 459) who sustained an eligible stroke were evaluated prospectively using standardized assessments at enrollment (within 14 days of stroke onset, 8.8 +/- 3.5 days). MAIN OUTCOME MEASURES: Mobility and activities of daily living (ADLs) were assessed at 1, 3, and 6 months poststroke using the Functional Independence Measure, Barthel index, Lawton Instrumental Activities of Daily Living (IADL), and the Medical Outcomes Study Short-Form Health Survey instruments. RESULTS: The cumulative probability of achieving independence with walking, a Barthel index of 60 or greater or 90 or greater, and independence in 3 or more IADL was significantly different for the following 4 impairment groups in descending order: motor; motor and somatosensory; motor and hemianopia; and motor, sensory, and hemianopia. Although motor severity was a strong predictor of outcome (p < .0001), the additional somatosensory and hemianopia deficits significantly (p < .05) affected time and likelihood of achieving these levels of function. CONCLUSION: Cumulative deficits poststroke affect patients' functional outcome in the first 6 months poststroke beyond the effect of motor severity alone. PMID- 11030502 TI - Quantification of pressure relief using interface pressure and tissue perfusion in alternating pressure air mattresses. AB - OBJECTIVE: To examine whether the interface pressure (IP) relief provided by alternating pressure air mattresses (APAMs) is matched with maintenance of tissue perfusion over the points of contact by measuring transcutaneous oxygen and carbon dioxide (tcPO2, tcPCO2). DESIGN: Comparative analysis of 2 APAMs with a 2 parameter continuous time-based method for quantifying pressure relief (PR) and transcutaneous gas measurement for assessing tissue perfusion. SETTING: Rehabilitation research facility in a university hospital. PARTICIPANTS: Eleven able-bodied adult postgraduate student volunteers. MAIN OUTCOME MEASURES: Two full-replacement APAM systems were used. For each mattress the mean maximum and minimum interface pressures; mean peak air pressures in the mattresses; interface pressure durations below 30, 20, and 10 mmHg over a 60-minute period; mean maximum tcPCO2 and minimum tcPO2; and mean area under the tcPO2 and tcPCO2 curves were measured for each subject. RESULTS: IP on the sacrum was held below thresholds of 30, 20, and 10 mmHg longer on a 2-cell, low pressure system than on a 3-cell, high pressure system (p < .001). Integrated over time, tcPO2 levels also indicated that the 2-cell system retained oxygen levels closer to the unloaded baseline than did the 3-cell system (p < .01). tcPCO2 levels did not rise significantly (p > 0.1) compared with the baseline measurement in both mattresses. CONCLUSIONS: PR was sensitive to the design of the APAM, especially its inflation pressure, cycle time, and inflation sequence. If future trials demonstrate that PR values and transcutaneous blood gas measurements correlate significantly with the clinical incidence of pressure sore formation, then this technique may prove useful in assessing the effectiveness of alternating pressure support surfaces. PMID- 11030503 TI - Muscle activation in therapeutic exercises to improve trunk stability. AB - OBJECTIVE: To evaluate the relative activation amplitudes from 3 abdominal and 2 trunk extensor muscle sites in healthy subjects performing the pelvic tilt, abdominal hollowing, and level 1 of the trunk stability test (TST level 1) exercises and to compare the activation amplitudes among muscle sites and exercises. DESIGN: Prospective comparative study. SETTING: Motion Analysis Research Center, Dalhousie University, Canada. PARTICIPANTS: Twenty-four healthy men (mean age, 30 +/- 8.1 yr [SD]) without low back pain. INTERVENTIONS: Subjects performed 3 exercises in a balanced order, repeating each exercise 5 times while surface electromyography (EMG) was recorded from the 5 muscle sites. Exercises were divided into 2 phases: movement and stabilization. The root-mean-square amplitude of the EMG for each phase was calculated and normalized to the maximal voluntary isometric contraction (MVIC) amplitude for each muscle. A 2-factor repeated-measures analysis of variance (ANOVA) tested the muscle by exercise interaction and the main effects for each phase separately. MAIN OUTCOME MEASURES: Normalized activation amplitude was the main dependent variable. Ensemble-average curves were calculated to examine the phasing of activation. RESULTS: ANOVA showed a statistically significant interaction (p < .05) for both phases, indicating the 3 exercises recruited the 5 muscle sites using different patterns of relative activation. The external oblique (EO) muscle site was activated to higher amplitudes than the other 4 sites in all 3 exercises for both phases. The highest activity was recorded from the EO during the pelvic tilt, just more than 25% of MVIC. The only exercise to recruit the erector spinae to significantly higher amplitudes than the multifidus site was the TST level 1. CONCLUSIONS: Study exercises were not interchangeable for the patterns of trunk muscle activation amplitudes. The exercises did not recruit the abdominal muscles to adequate levels for strengthening for this healthy sample; however, all 5 muscle sites were activated, forming the basis of a stabilizing exercise approach. PMID- 11030504 TI - Secondary conditions and women with physical disabilities: a descriptive study. AB - OBJECTIVE: To examine secondary conditions and their relationship to life satisfaction in women with physical disabilities. DESIGN: Survey research. SETTING: Metropolitan Philadelphia. PARTICIPANTS: Women aged 18 to 65 years with self-reported functional limitations associated with neurologic, neuromuscular, brain, sensory, arthritic, spinal cord, orthopedic, or other chronic conditions. MAIN OUTCOME MEASURES: Secondary conditions, health status, functional status, and life satisfaction. RESULTS: In the past year, an average of 12 +/- 6.1 (SD) secondary conditions were experienced. Fatigue, mobility, physical deconditioning, spasticity, and joint pain were reported most frequently, followed by depression, chronic pain, access problems, weight problems, and isolation. In comparing women with multiple sclerosis (MS) and those with other disabling conditions, women with MS had higher secondary conditions problem index (SCPI) scores for bladder, bowel, and sexual dysfunction than women in the mixed disability group. Other secondary conditions with the highest SCPI scores remained similar. Self-reported emotional health status and SCPI scores were the only significant predictors of life satisfaction. CONCLUSION: Secondary conditions influence health status and quality of life for women with physical disabilities. Despite differences among disability groups in terms of the type of secondary conditions, common secondary conditions experienced by most women may be amenable to non-disability-specific health promotion programs. PMID- 11030505 TI - Validity of an artificial neural network in predicting discharge destination from a postacute geriatric rehabilitation unit. AB - OBJECTIVE: To develop an artificial neural network (ANN) designed to predict discharge destination from postacute geriatric rehabilitation units. DESIGN: Nonconcurrent prospective study. SETTING: Postacute geriatric rehabilitation units: a 20-bed unit in a nonproprietary skilled nursing facility and a 40-bed unit in a suburban private facility. PATIENTS: Consecutive sample of 661 patients admitted between January 1995 and February 1999, including a derivation group of 452 patients and a validation group of 209 patients. INTERVENTIONS: A feed forward, back-propagation neural network to predict discharge destination. MAIN OUTCOME MEASURE: Discharge destination from postacute geriatric rehabilitation. RESULTS: An ANN was trained on clinical pattern set derived from 452 patients and validated prospectively on 209 consecutive patients admitted to postacute geriatric rehabilitation units. The neural network achieved a sensitivity of 85.7% (95% confidence interval [CI], 83.7-89.4) and specificity of 94.1% (95% CI, 84.4-99.1) in identifying discharge destination with a corresponding area under the curve of 95.7% (95% CI, 92.1-98.3). CONCLUSION: An ANN can predict discharge to the community postacute rehabilitation with a high degree of accuracy. It could have particular value to predict return to the community for older adults with multiple comorbidities after an acute hospitalization. PMID- 11030506 TI - Disability measurement in persons with back pain: a validity study of spinal range of motion and velocity. AB - OBJECTIVE: To evaluate the criterion validity and responsiveness to change of spine kinematic variables to assess disability in patients with low back pain. DESIGN: Blinded comparison between spine kinematic variables, Oswestry disability questionnaire scores, and work status. SETTING: Multidisciplinary occupational rehabilitation clinic of a university hospital. PATIENTS: Population-based cohort of 111 patients with subacute work-related back pain who were absent from regular work for more than 4 weeks because of back pain. INTERVENTIONS: This study was part of a population-based randomized clinical trial. Patients were randomized to 4 different methods of management: usual care, rehabilitation, ergonomics, or rehabilitation and ergonomics. MAIN OUTCOME MEASURES: Oswestry disability questionnaire, kinematic analysis of the spine during flexion and extension of the trunk, and work status were collected at weeks 4, 12, 24, and 52 after the back accident. RESULTS: Kinematic variables were poorly to moderately related to work status and Oswestry questionnaire scores. Kinematic variables were also unresponsive to change in work status and Oswestry questionnaire scores over time. CONCLUSION: Spine kinematics during flexion and extension of the trunk do not appear to be a valid measure of disability in patients with subacute and chronic back pain. PMID- 11030507 TI - An objective and standardized test of foot function: normative values and validation in patients with reflex sympathetic dystrophy. AB - OBJECTIVE: To describe and obtain normative values for an objective and standardized test of foot function, and to validate the test in patients with impaired function of 1 leg. DESIGN: A series of 4 standardized and objectively measured subtests, representing common foot activities in daily living, was devised. SETTING: University hospital. PARTICIPANTS: Normative values were obtained for 100 healthy patients between 20 and 70 years of age. For validation purposes, the test was also performed by 20 patients diagnosed with reflex sympathetic dystrophy of 1 foot. MAIN OUTCOME MEASURES: Several basic aspects of individual foot function were evaluated: (1) forward and backward shifting (FBS) of a foot panel; (2) lateral shifting (LS) of a foot panel; (3) alternately touching 2 bells (TB); and (4) depressing a pedal (DP). (The tests were performed while seated; hence, they are applicable to patients unable to walk.) Comparison with results on a battery of other clinical function tests was assessed. RESULTS: The intrarater and interrater reliabilities of the test were high (eg, intrarater correlation coefficients ranged from .74 to .93; interrater from .85 to .99). Results were influenced by sex and dominance, but were not influenced by height, weight, or shoe size. Age and leisure activities involving foot function influenced 1 subtest only (depressing a pedal). Results of the affected side in patients were higher than normative values although, to a lesser extent, the same was true for the unaffected side. Footboard results did not correlate with results of other function tests, except myometry, suggesting that it provides additional information. In contrast to the other tests, and like myometry, the footboard distinguished patients who were crutch dependent from those who were not. CONCLUSION: The footboard is a valuable addition to current tests for assessing foot function. PMID- 11030508 TI - Motor control testing of upper limb function after botulinum toxin injection: a case study. AB - OBJECTIVE: To evaluate changes in upper extremity function in a hemiparetic patient after treatment with botulinum toxin (BTX) using motor-control testing (MCT) techniques. DESIGN: Interventional with longitudinal study, open label. SETTING: A children's hospital and a motor-control laboratory at a major academic center. PARTICIPANTS: A 16-year-old male with right hemiparetic cerebral palsy and a healthy 12-year-old control subject. INTERVENTIONS: BTX injections to the elbow and wrist flexors. MAIN OUTCOME MEASURES: MCT was used to examine 4 upper extremity movements: forward reach, bilateral rhythmic movements (both muscle homologous and direction homologous), isometric pinch, and hand tapping. The patient was tested before treatment and at 2, 4, 6, 12, 18, and 24 weeks after treatment. In addition, range of motion (ROM), the Ashworth scale of spasticity, Functional Independence Measure, and the mobility and activities of daily living (ADL) sections of the Pediatric Evaluation of the Disability Inventory were performed. RESULTS: Forward reach demonstrated little change initially despite patient reports of "feeling looser." Improvement was noted after 18 weeks, but returned to baseline level at 24 weeks. Bilateral rhythmic movements also showed slight improvement at 18 weeks. Pinch force increased significantly after 2 weeks, but declined again at 6 weeks. Improvements occurred in ROM and the Ashworth rating of spasticity, but were not temporally associated with each other or with MCT results. Functional assessment data did not change during the study period. CONCLUSIONS: Improvements in more complex motor tasks were noted after significant delay from the time of treatment, while simpler tasks demonstrated a more rapid improvement, followed by a rapid return to baseline levels. This case suggests that MCT techniques can provide quantitative and qualitative data, which can add new information about upper extremity motor disability and the outcome of treatment. PMID- 11030509 TI - Treatment of proximal plantar fasciitis with ultrasound-guided steroid injection. AB - OBJECTIVE: To investigate the efficacy of ultrasound-guided steroid injection for the treatment of proximal plantar fasciitis and to evaluate mechanical properties of the heel pad after steroid injection. DESIGN: Proximal plantar fascia and heel pad were assessed with a 10-MHz linear array ultrasound transducer. Pain intensity was quantified with a tenderness threshold (TT) and visual analog scale (VAS). The transducer was incorporated into a specially designed device to measure mechanical properties of the heel pad. Evaluations were performed before injection and at 2 weeks and 3 months after injection. SETTING: An outpatient clinic of a tertiary care center. PATIENTS: Fourteen consecutive patients with unilateral proximal plantar fasciitis. INTERVENTION: Ultrasound-guided injection of 7 mg betamethasone and 0.5 mL of 1% lidocaine into the inflamed proximal plantar fascia. MAIN OUTCOME MEASURES: VAS, TT, heel pad and plantar fascia thickness, and echogenicity of the proximal plantar fascia on sonogram were assessed. Mechanical properties included unloaded heel pad thickness, compressibility index, and energy dissipation ratio. RESULTS: Both VAS score +/- standard deviation (SD; 5.43 +/- 2.03, 1.39 +/- 2.19, 0.57 +/- 1.40 at the 3 measurements, respectively) and TT +/- SD (5.05 +/- 1.42, 9.34 +/- 1.84, 9.93 +/- 1.98 kg/cm2 at the 3 measurements, respectively) improved significantly (p < .001) after steroid injection. The mean thickness of the plantar fascia was greater in the symptomatic side than in the asymptomatic side before treatment (0.58 +/- 0.13 cm vs 0.40 +/- 0.11 cm, p < .001). The thickness had decreased significantly 3 months after injection (0.46 +/- 0.12 cm at 2 weeks, 0.42 +/- 0.10 cm at 3 months, p < .001). The hypoechogenicity at the proximal plantar fascia disappeared after steroid injection (p < .001). Mechanical properties of the heel pad did not change 3 months after steroid injection (p > .05). CONCLUSION: Ultrasound offers an objective measurement of the therapeutic effect on proximal plantar fasciitis. Accurate steroid injection under ultrasound guidance can effectively treat proximal plantar fasciitis without significant deterioration of the mechanical properties of the heel pads. PMID- 11030510 TI - The relationship between self-reports and objective measures of disability in patients with late sequelae of poliomyelitis: a validation study. AB - OBJECTIVE: To establish the validity of 2 objective measures of disability based on 2 self-reports in individuals with the late sequelae of poliomyelitis (LSP). DESIGN: Nonexperimental research design. SETTING: Regional ambulatory clinic specializing in the management of patients with LSP. PARTICIPANTS: Seventeen community dwellers with LSP. INTERVENTION: Completion of 2 exercise tests: the 6 minute walk (6-MW) test and a steady-rate walking test to calculate the cardiorespiratory conditioning index (CRCI), and 2 health status questionnaires: the Sickness Impact Profile (SIP) and Medical Outcomes Study Short-Form 36 (SF 36). MAIN OUTCOME MEASURES: 6-MW distance and the CRCI, and scores from the physical dimensions of the SIP and the SF-36. RESULTS: The 6-MW distance correlated significantly with the physical dimension scores of both the SIP (r = .57, p < .05) and the SF-36 (r = .67, p < .05). Significant correlations were also observed between the CRCI and the physical dimension scores of the SIP (r = .51, p < .05) and the SF-36 (r = .70, p < .01). The 2 objective measures correlated significantly with each other (r = .51, p < .05) as did the 2 physical dimension scores of the SIP and the SF-36 (r = -.80, p < .01). CONCLUSIONS: Although objective measures of disability corroborated self-reports of patients with LSP and, thus, supported their validity, each measure has a distinct role in clinical decision making. PMID- 11030511 TI - Intrarater reliability of manual passive movement velocity in the clinical evaluation of knee extensor muscle tone. AB - OBJECTIVE: To quantify the relationship between therapist-applied velocity of passive movement during a manual muscle test of muscle tone and the level of muscle tone represented by the relaxation index of the pendulum test. DESIGN: Comparison of therapist-applied passive limb movement velocity during a manual muscle test with the same subject's level of muscle tone measured by the pendulum test. Three different therapists tested each subject. The relation between the velocity scores and pendulum test scores both intratherapist and intertherapist were assessed statistically by means of analysis of variance and correlation coefficients. SETTING: A university-affiliated tertiary care outpatient and inpatient spinal cord injury rehabilitation center. PARTICIPANTS: Twenty-two volunteer subjects with spinal cord injuries. MAIN OUTCOME MEASURES: Passive knee angular displacement data were collected during both manual knee muscle testing and pendulum tests by using an electrogoniometer. RESULTS: The therapists produced significantly different movement velocities during the manual muscle tests (p < .05). A significant correlation (p < .001) was found between pendulum test scores and passive velocities, indicating that in higher levels of muscle tone, the greater stretch reflex present provided greater resistance against the therapist force and subsequently decreased the velocity of the passive stretching. CONCLUSIONS: Measurement of average velocity during passive stretching by itself can be used to evaluate muscle tone. PMID- 11030512 TI - Alcohol neurolysis of the sciatic nerve in the treatment of hemiplegic knee flexor spasticity: clinical outcomes. AB - OBJECTIVE: To study clinical outcomes after 50% to 100% alcohol neurolysis of the sciatic nerve in the treatment of chronic hemiplegic knee flexor spasticity. DESIGN: Case series using a convenience sample. SETTING: Rehabilitation teaching hospital. PATIENTS: Eight patients (mean age, 55.1 +/- 12.1 yr; mean duration to block, 4.4 +/- 3.7 mo) with chronic hemiplegia and severe spasticity (modified Ashworth scale [MAS] score < 2) secondary to cerebrovascular accidents (n = 5) and traumatic brain injuries (n = 3). Two patients were ambulant, 6 were wheelchair-bound. INTERVENTIONS: Fifty percent to 100% alcohol neurolysis of the sciatic nerve using repetitive electric stimulation to localize the sciatic nerve. MAIN OUTCOME MEASURES: MAS score, gain in knee range of motion (ROM), and visual inspection of gait at 0, 1, and 6 months postneurolysis. Nonparametric tests were used in statistical analysis. RESULTS: The mean preinjection MAS score was 2.8 +/- 0.7, which improved to 1.4 +/- 0.7 at 1 month (p = .005), and 1.8 +/- 0.9 (p = .01) and 1.9 +/- 1.1 (p = .02) at 3 and 6 months postinjection, respectively. The mean gain in knee ROM was 34.4 degrees +/- 15.7 degrees at 1 month postinjection and was maintained at 6 months postinjection. Improvements in gait were noted in both ambulant patients, and improved positioning in 3 of 6 wheelchair-bound patients. The incidence of dysesthetic pain was 0%. CONCLUSION: Fifty percent to 100% alcohol neurolysis of the sciatic nerve is a safe and effective method for treatment of hemiplegic knee flexor spasticity, with therapeutic effects lasting 6 months. PMID- 11030513 TI - Painless aortic dissection presenting as high paraplegia: a case report. AB - Acute aortic dissection is a catastrophic episode that usually presents as a sudden, painful, ripping sensation in the chest or back. Physical findings may include loss of pulses and aortic regurgitation. It is associated with neurologic sequelae in as many as one third of patients. Painless dissection occurs in 5% of patients. We report a case of painless aortic dissection, presenting as acute paraplegia. The patient was a 77-year-old woman who presented with paraplegia, with no chest or back pain. On examination, strength was 5/5 in both upper extremities and 0/5 in both lower extremities. Deep tendon reflexes were absent in her legs. She had no voluntary anal contraction. Sensation was absent from T6 through S5. Computed tomography of the chest revealed a type A dissecting aneurysm. The vascular supply to the spinal cord and the differential diagnosis for new onset paraplegia are discussed. PMID- 11030514 TI - Gabapentin for chronic pain in spinal cord injury: a case report. AB - A 30-year-old white woman with an L1 complete spinal cord injury (SCI) secondary to a gunshot wound in 1985, presented to a chronic pain service for evaluation. She had a 13-year history of chronic lower extremity pain. She described her discomfort as "throbbing, aching, and stabbing." She had tried many different medications, including opioids, Tegretol, and tricyclic antidepressants, without success. During the evaluation process, she admitted to being "angry, frustrated, and anxious." She was diagnosed with central pain after SCI. She was placed on gabapentin 300 mg 3 times daily; within 1 week, her visual analog pain scale fell from 95 mm to 27 mm, and her McGill Short Form pain score fell from 13 to 3. Her mood also vastly improved. This case report suggests that gabapentin should be studied as a therapeutic option for treating central pain post-SCI and should be considered as a viable, well-tolerated, low-toxicity tool. PMID- 11030515 TI - Superior mesenteric artery syndrome in traumatic paraplegia. PMID- 11030516 TI - Make tests and proper muscle strength measurement. PMID- 11030517 TI - Renewal-process approximation of a stochastic threshold model for electrical neural stimulation. AB - In a recent set of modeling studies we have developed a stochastic threshold model of auditory nerve response to single biphasic electrical pulses (Bruce et al., 1999c) and moderate rate (less than 800 pulses per second) pulse trains (Bruce et al., 1999a). In this article we derive an analytical approximation for the single-pulse model, which is then extended to describe the pulse-train model in the case of evenly timed, uniform pulses. This renewal-process description provides an accurate and computationally efficient model of electrical stimulation of single auditory nerve fibers by a cochlear implant that may be extended to other forms of electrical neural stimulation. PMID- 11030518 TI - Subthreshold voltage noise due to channel fluctuations in active neuronal membranes. AB - Voltage-gated ion channels in neuronal membranes fluctuate randomly between different conformational states due to thermal agitation. Fluctuations between conducting and nonconducting states give rise to noisy membrane currents and subthreshold voltage fluctuations and may contribute to variability in spike timing. Here we study subthreshold voltage fluctuations due to active voltage gated Na+ and K+ channels as predicted by two commonly used kinetic schemes: the Mainen et al. (1995) (MJHS) kinetic scheme, which has been used to model dendritic channels in cortical neurons, and the classical Hodgkin-Huxley (1952) (HH) kinetic scheme for the squid giant axon. We compute the magnitudes, amplitude distributions, and power spectral densities of the voltage noise in isopotential membrane patches predicted by these kinetic schemes. For both schemes, noise magnitudes increase rapidly with depolarization from rest. Noise is larger for smaller patch areas but is smaller for increased model temperatures. We contrast the results from Monte Carlo simulations of the stochastic nonlinear kinetic schemes with analytical, closed-form expressions derived using passive and quasi-active linear approximations to the kinetic schemes. For all subthreshold voltage ranges, the quasi-active linearized approximation is accurate within 8% and may thus be used in large-scale simulations of realistic neuronal geometries. PMID- 11030519 TI - Evidence for a distinct light-induced calcium-dependent potassium current in Hermissenda crassicornis. AB - A model of phototransduction is developed as a first step toward a model for investigating the critical interaction of light and turbulence stimuli within the type B photoreceptor of Hermissenda crassicronis. The model includes equations describing phototransduction, release of calcium from intracellular stores, and other calcium regulatory mechanisms, as well as equations describing ligand gating of a rhabdomeric sodium current. The model is used to determine the sources of calcium in the soma, whether calcium or IP3 is a plausible ligand of the light-induced sodium current, and whether the light-induced potassium current is equivalent to the calcium-dependent potassium current activated by light induced calcium release. Simulations show that the early light-induced calcium elevation is due to influx through voltage-dependent channels, whereas the later calcium elevation is due to release from intracellular stores. Simulations suggest that the ligand of the fast, light-induced sodium current is IP3 but that there is a smaller, prolonged component of the light-induced sodium current that is activated by calcium. In the model, the calcium-dependent potassium current, located in the soma, is activated only slightly by light-induced calcium elevation, leading to the prediction that a calcium-dependent potassium current, active at resting potential, is located in the rhabdomere and is responsible for the light-induced potassium current. PMID- 11030520 TI - The autapse: a simple illustration of short-term analog memory storage by tuned synaptic feedback. AB - According to a popular hypothesis, short-term memories are stored as persistent neural activity maintained by synaptic feedback loops. This hypothesis has been formulated mathematically in a number of recurrent network models. Here we study an abstraction of these models, a single neuron with a synapse onto itself, or autapse. This abstraction cannot simulate the way in which persistent activity patterns are distributed over neural populations in the brain. However, with proper tuning of parameters, it does reproduce the continuously graded, or analog, nature of many examples of persistent activity. The conditions for tuning are derived for the dynamics of a conductance-based model neuron with a slow excitatory autapse. The derivation uses the method of averaging to approximate the spiking model with a nonspiking, reduced model. Short-term analog memory storage is possible if the reduced model is approximately linear and if its feedforward bias and autapse strength are precisely tuned. PMID- 11030521 TI - Electrophysiological and theoretical analysis of depolarization-dependent outward currents in the dendritic membrane of an identified nonspiking interneuron in crayfish. AB - Depolarization-dependent outward currents were analyzed using the single electrode voltage clamp technique in the dendritic membrane of an identified nonspiking interneuron (LDS interneuron) in situ in the terminal abdominal ganglion of crayfish. When the membrane was depolarized by more than 20 mV from the resting potential (65.0 +/- 5.7 mV), a transient outward current was observed to be followed by a sustained outward current. Pharmacological experiments revealed that these outward currents were composed of 3 distinct components. A sustained component (I(s)) was activated slowly (half rise time > 5 msec) and blocked by 20 mM TEA. A transient component (I(t1)) that was activated and inactivated very rapidly (peak time < 2.5 msec, half decay time < 1.2 msec) was also blocked by 20 mM TEA. Another transient component (I(t2)) was blocked by 100 microM 4AP, activated rapidly (peak time < 10.0 msec) and inactivated slowly (half decay time > 131.8 msec). Two-step pulse experiments have revealed that both sustained and transient components are not inactivated at the resting potential: the half-maximal inactivation was attained at -21.0 mV in I(t1), and 38.0 mV in I(t2). I(s) showed no noticeable inactivation. When the membrane was initially held at the resting potential level and clamped to varying potential levels, the half-maximal activation was attained at -36.0 mV in I(s), -31.0 mV in I(t1) and -40.0 mV in I(t2). The activation and inactivation time constants were both voltage dependent. A mathematical model of the LDS interneuron was constructed based on the present electrophysiological records to simulate the dynamic interaction of outward currents during membrane depolarization. The results suggest that those membrane conductances found in this study underlie the outward rectification of the interneuron membrane as well as depolarization dependent shaping of the excitatory synaptic potential observed in current-clamp experiments. PMID- 11030522 TI - Thromboembolic prophylaxis with danaparoid (Orgaran) in a high-thrombosis-risk pregnant woman with a history of heparin-induced thrombocytopenia (HIT) and Widal's disease. AB - There is no consensus concerning thromboembolic prophylaxis in high-risk pregnant women with a previous history of heparin-induced thrombocytopenia. An alternative anticoagulant therapy is danaparoid, whereas unfractioned and low-molecular weight heparin therapy is contraindicated. We report a case of successful thrombosis prophylaxis using danaparoid in a high-thrombosis-risk pregnant woman with a history of heparin-induced thrombocytopenia during a previous pregnancy and Widal's disease. PMID- 11030523 TI - Differential effects of unfractionated heparin and low-molecular-weight heparins on tissue thromboplastin inhibition test. AB - Circulating anticoagulants are endogenously produced substances that interfere with in vitro tests of coagulation like activated partial thromboplastin time (APTT), and cause prolongation of the clotting times. Evaluation of the abnormal APTT involving various factor assays and mixing studies may provide inconclusive and ambiguous results. Tissue thromboplastin inhibition test (TTIT) is one of the screening assays for detection of circulating anticoagulants. However, this test is influenced by the presence of unfractionated heparin (UFH) from concentrations 0.2 U/mL and higher. Since low-molecular-weight heparins (LMWHs) are increasingly used for the prevention of thrombotic disorders and may replace UFH in the future, in this study the authors studied the influence of LMWHs on the performance of TTIT and compared the results with UFH. UFH and LMWHs showed a prolongation of TTIT in the concentration range of 0.25-1.0 U/mL. The marked prolongation of the TTIT with UFH and different LMWHs is in decreasing order of UFH > ardeparin > tinzaparin > dalteparin > enoxaparin. Patients with circulating anticoagulants who are given LMWHs may have false-positive results of TTIT and this influence should be kept in mind during patient management. PMID- 11030524 TI - Changes of hemostatic molecular markers after gynecological surgery. AB - The authors evaluated the hemostatic abnormalities occurring in the postoperative period of eight patients with malignant tumors and compared them with those occurring in the postoperative period of eight patients with benign tumors. Two of the patients with malignant tumor presented pulmonary embolism after operation. Plasma fibrinogen and fibrin degradation product levels in patients with malignant tumors were already high before operation and further increased significantly after operation. The plasma levels of D-dimer, thrombin antithrombin complex, and free-tissue factor pathway inhibitor were increased in both groups after operation, but they were higher in patients with malignant tumors than in patients with benign tumors. The plasma levels of protein C and antithrombin were significantly decreased in both groups after operation. but they were significantly lower in patients with malignant tumors than in those with benign tumors. The decreased activity of protein C or antithrombin may be not only a risk factor of thrombotic disease, such as pulmonary embolism, but also the cause of thrombosis. In patients with malignant tumors, the operation time was significantly longer than that in patients with benign tumors. This long operative period might cause vascular endothelial cell injury which is reflected by the plasma levels of free-tissue factor pathway inhibitor, antithrombin, and protein C. PMID- 11030525 TI - Decreased protein C activation is associated with abnormal collagen turnover in the intraalveolar space of patients with interstitial lung disease. AB - Activation of the coagulation system in the alveolar space plays an important role in the pathogenesis of interstitial lung disease (ILD) and pulmonary fibrosis. The protein C (PC) pathway is the main modulator of coagulation activation. This study evaluated whether dysfunction of the PC pathway is associated with increased collagen synthesis in the intraalveolar space of patients with ILD. This study comprised 22 patients with ILD; of these, five had idiopathic pulmonary fibrosis (IPF), nine had sarcoidosis-associated ILD, and eight had collagen vascular disease-associated ILD (CVD-ILD). Thrombin antithrombin complex (TAT) was measured as a marker of coagulation activation. As markers of the PC pathway activity, the concentration of activated PC-PC inhibitor (APC-PCI) complex and the APC-PCI/PC ratio were measured and, as a marker of collagen synthesis, the concentration of aminoterminal propeptide of type III procollagen (PIIINP) was measured in bronchoalveolar lavage fluid (BALF) of ILD patients. TAT was significantly increased in BALF from ILD patients as compared to control subjects. The concentrations of PIIINP were significantly elevated in patients with ILD as compared to healthy subjects. In contrast, the concentration of APC-PCI and the values of APC-PCI/PC ratio were significantly decreased in BALF from patients with ILD. BALF concentration of PIIINP was significantly and inversely correlated with the concentration of APC-PCI and with the APC-PCI/PC ratio. These findings suggest that dysfunction of the protein C pathway may have important physiopathologic implications in the development of pulmonary fibrosis in ILD. PMID- 11030526 TI - Plasma tissue factor pathway inhibitor levels as a marker for postoperative bleeding after enoxaparin use in deep vein thrombosis prophylaxis in orthopedics and general surgery. AB - Low-molecular-weight heparins (LMWH) are widely used as antithrombotic prophylactic pharmaceutical agents in orthopedic and general surgery. Their antithrombotic characteristics are expressed by plasma mediators such as anti-Xa, anti-IIa, and increased release of tissue factor pathway inhibitor (TFPI) from vascular endothelium. The purpose of this clinical research is to study the relation between plasma levels of these mediators and postoperative bleeding. Forty-one consecutive patients undergoing hip or knee arthroplasty (n = 36) and colectomy (n = 5) received the standard enoxaparin (a LMWH) dose preoperatively (general surgery) or immediately postoperatively (orthopedic surgery). Major bleeding was defined as a postoperative drop of > or = 5 g/dL) of hemoglobin. The authors observed that there was a linear relationship between an increase in free/total TFPI ratio levels and postoperative bleeding. When that ratio increased by > 60%, the hemoglobin dropped to > 5 g/dL (n = 17). This relationship between free/total TFPI ratio increase and postoperative bleeding was statistically significant (P < 0.001). Those who did not bleed (hemoglobin drop was less than 5 g/dL) (n = 24) had a ratio increase (if any) of less than 50%. However, the authors did not observe any statistical relationship between anti-Xa, anti-IIa, or prothrombin time and postoperative bleeding in patients receiving LMWH for deep vein thrombosis prophylaxis in orthopedic and general surgery patients. The authors recommend a pre- and postoperative ratio level measurement whenever major bleeding is anticipated, as adjustments of LMWH dose or frequency might be necessary. PMID- 11030527 TI - Platelets expressing P-selectin and platelet-derived microparticles in stored platelet concentrates bind to PSGL-1 on filtrated leukocytes. AB - The levels of interleukin-6 and platelet-derived microparticles (PMPs) were measured in the blood of 137 patients with side effects from platelet concentrate (PC) transfusion with leukocyte removal filtration, P-selectin-expressing platelet and PMPs in stored PC before and after the filtration, and filtered leukocytes positive for P-selectin glycoprotein ligand-1. The side effects, which were observed in 203 transfusions for 84 patients with hematologic disease and 53 patients with nonhematologic disease with no significant difference between the two groups, included urticaria (75.9%), erythema (18.7%), and fever (17.2%), but no anaphylactic reactions. The levels of interleukin-6 and PMP correlated in both groups, and were significantly higher in the hematologic disease group than in the nonhematologic disease group. The level of PMP, but not interleukin-6, was significantly higher for patients testing positive for allergic reaction than for those testing negative. In the stored PC prior to filtration, the level of interleukin-6 was normal. The level of P-selectin-expressing platelets and PMPs was elevated before filtration, but was significantly lower after filtration. Taken together, the results suggest that PMP is involved in the generation of transfusion reactions, and indicate that both platelets and PMP displaying P selectin bind to P-selectin glycoprotein ligand-1 of leukocytes retained by the leukocyte filter. PMID- 11030528 TI - Ticlopidine, Alka-Seltzer, or a combination of citric acid with aspirin: effects on platelet aggregation in individuals with an insufficient response to aspirin alone. AB - Aspirin (ASA) does not effectively lower platelet aggregation in all people. The platelet aggregation (PA) score is an easily used clinical method for measuring the effect in individuals of antiplatelet medications. Fifteen apparently healthy subjects (2 men and 13 women), selected for their resistance to ASA's antiaggregation effect, completed a sequential trial of ticlopidine, Alka Seltzer, and ASA + citric acid (CTA). Ticlopidine was the strongest aggregation inhibitor and the ASA + CTA combination was more inhibitory than Alka-Seltzer. It was determined that measuring antiaggregation effects of a particular agent in an individual prior to usage would optimize treatment. The PA score methodology provides a means for testing patients prior to antiplatelet therapy for prevention and treatment of the thrombotic complications of vascular disease. PMID- 11030529 TI - Thromboelastograph assay for measuring the mechanical strength of fibrin sealant clots. AB - In order to provide sustained hemostasis or tissue sealing, fibrin sealants must generate adhesive clots with mechanical properties capable of resisting forces, such as shear, that might break or tear the clot. Commercial preparations of fibrin sealants should generate clots of adequate and consistent mechanical strength. The mechanical strength of fibrin sealants is often measured as bonding strength in in vivo or ex vivo animal wound models. These tests can be useful predictors of clinical efficacy. However, these, as well as many in vitro tensile strength tests for fibrin sealant, tend to be laboratory specific and require extensive reagent preparation time and analyst training. The thromboelastograph has historically been used to screen for plasma protein and platelet disorders that lead to defective clot formation. The authors have developed a simple in vitro test, using a standard thromboelastograph that can provide reliable, reproducible information on the rheology of clots generated by fibrin sealant preparations. Using this method, the shear strength of fibrin sealant clots was measured and shown to correlate with the fibrinogen, but not the thrombin, concentration in the sealant. Shear strength was also shown to correlate with the sealant concentration of the fibrin cross-linking proenzyme, factor XIII. Sealants containing lysine, which can act as an alternate substrate for factor XIII enzyme and prevent efficient fibrin chain cross-linking, were shown by this method to generate clots of substantially reduced shear strength. The method distinguished between thrombin-catalyzed clot formation and other fibrinogen clotting mechanisms as evidenced by the significantly lower shear strength associated with batroxobin-generated fibrin clots. PMID- 11030530 TI - L-asparaginase-provoked seizures as singular expression of central nervous toxicity. AB - Patients treated with L-asparaginase may present with hemorrhagic and thrombotic cerebrovascular events. This syndrome generally occurs after a few weeks of therapy and may occur after L-asparaginase therapy is completed. Complications appear to result from depletion of plasma proteins involved in coagulation and fibrinolysis. Seizures are uncommon symptoms, and are always caused by cerebrovascular events. We report a case of seizure associated with L asparaginase therapy but no evidence of hemorrhagic or thrombotic cerebrovascular events. PMID- 11030531 TI - The significance or nonsignificance of the G to A 20210 prothrombin polymorphism. PMID- 11030532 TI - Controversial cases in endourology. PMID- 11030533 TI - Shockwave frequency affects fragmentation in a kidney stone model. AB - PURPOSE: To study the effect of altering shockwave frequency on the efficiency of stone fragmentation using the MFL 5000 spark-gap lithotripter. MATERIALS AND METHODS: Standardized solid plaster stones, 12.0 +/- 0.5 mm in diameter, were fragmented at an energy setting of 20 kV. The shockwave frequencies tested were 60, 80, and 117 per minute. Stones were fragmented throughout the entire lifespan of the electrode, from 0 to >100% consumption, at each frequency tested. Electrode pressure output was studied for each frequency. RESULTS: A greater number of shocks was required to fragment the plaster balls at higher frequencies (regression coefficient 1.93; p < 0.003). An inverse relation was found between the number of shocks necessary to break the stones and electrode consumption (regression coefficient -2.16; p < 0.001). The analysis of delivered pressure from the electrode failed to demonstrate a linear relation with frequency (regression coefficient -0.40; p < 0.728) or consumption (regression coefficient 1.11; p < 0.158). CONCLUSIONS: The number of shocks required to fragment a stone is influenced in part by the frequency at which the shockwaves are delivered. Increasing the shockwave frequency from 60 to 117 per minute in this study caused a significant rise in the number of shocks required to break the stone. The pressure output of the electrode was similar at the frequencies tested, thus making the difference in stone fragmentation secondary to the mechanism of stone disintegration and not the function of the electrode. PMID- 11030534 TI - Therapeutic efficacy of Dornier MPL 9000 for prevesical calculi as judged by efficiency quotient. AB - PURPOSE: To study the efficacy and safety of the Dornier MPL 9000 lithotripter in the treatment of prevesical calculi using real-time ultrasound monitoring and to see if efficacy is adequately judged by the efficiency quotient (EQ). PATIENTS AND METHODS: Seventy-six patients underwent ultrasound-guided extracorporeal shockwave lithotripsy (SWL) for prevesical calculi over a period of 43 months. Their age ranged from 8 to 68 (mean 37.6) years, and the male:female ratio was 2.6:1. The size of the stones, measured in two dimensions, ranged from 4 to 25 mm (mean 9.3 mm) and 2 to 15 mm (mean 6.0 mm). No general or regional anesthesia or ureteral stents were used, and all patients were treated in the prone position. The EQ was calculated using the formula: Stone free (%) x 100/(100 + retreatment rate (%) + auxiliary procedures (%). RESULTS: Seventy patients were stone free in a mean time of 15 +/- 14.11 days. The average number of shockwaves used was 2,831 +/- 1,612, and the average number of sessions per patient was 1.7. About 92% of the patients were rendered stone free using in situ SWL alone. No major complication was encountered, and none of the patients required an inpatient stay after SWL. The EQ was 65. Five patients with failed SWL subsequently were rendered stone free, four with salvage ureteroscopy and pneumatic lithotripsy and one with cystolitholapaxy for a symptomatic fragment in the bladder. One patient was lost to follow-up. CONCLUSION: In situ SWL is a safe and effective treatment for prevesical calculi. It should be used as a first-line treatment for most such stones. Efficiency can be assessed objectively by the EQ. PMID- 11030535 TI - CT analysis of caliceal anatomy in the supine and prone positions. AB - PURPOSE: To assess the effect of patient positioning on the position of the kidneys and their consequent projection onto plain radiographs, thus ascertaining the need for special preoperative imaging. PATIENTS AND METHODS: Fourteen patients were studied by obtaining fine (5-mm)-cut contrast-enhanced CT scans in the pyelogram phase in both the supine and prone position. The orientation of the kidneys relative to the midsagittal plane of the body and the orientation of the anterior and posterior calices relative to the axis of the kidney were measured from hardcopy images. Comparison was made between prone and supine positions for left and right kidneys separately, as well as overall. RESULTS: The position of the patient had a small effect on the orientation of the kidneys, with the mean angle changing from 56.6 degrees when supine to 61.6 degrees when prone (p < 0.05). However, no significant change in caliceal orientation or the relative projection of the anterior and posterior calices occurred as a result. CONCLUSION: There is no need to carry out special preoperative imaging in the operative or prone position. PMID- 11030536 TI - Outpatient ureteroscopic lithotripsy: selective internal stenting and factors enhancing success. AB - PURPOSE: To evaluate a policy of selective, short-duration internal stenting after outpatient ureteroscopic laser lithotripsy. PATIENTS AND METHODS: From January 1997 to April 1998, 62 patients (34 male, 28 female) with a mean age of 50 (range 21-80) years underwent outpatient ureteroscopic lithotripsy using a holmium laser (365 microm; 0.5-1.4J/5-10 Hz) and 6F/7.5F semirigid ureteroscope. Internal stents were inserted selectively in patients with severe preoperative obstruction (intravenous urogram finding), tight stone impaction (endoscopic finding), or significant residual obstruction (on-table retrograde pyelogram finding) despite stone clearance. Patient demographics, stone measures, stone clearance rates, complications, postoperative pain scores, analgesic requirement, and follow-up imaging were compared for the stented and unstented patients. RESULTS: With the present criteria of selective internal stenting, stents were inserted in 56% of the patients for a mean duration of 3.6 weeks. Excluding those patients with residual stones requiring further interventions, the stenting rate was 39% with a mean duration of 1.9 weeks. There was no difference in patient characteristics, stone burden, and stone levels between the stented and unstented group. The mean operating time for the unstented group was shorter than for the stented group (45.6 minutes v 56.6 minutes; P = 0.03). The stone clearance rates were similar for the two groups (96% v 97%), but the complication rate of the stented group was higher (8.6% v 3.7%). The mean postoperative pain score and analgesic requirement were similar in the two groups on postoperative day 1 but significantly less in the unstented group on day 3. CONCLUSIONS: The criteria for selective internal stenting are useful in determining when a stent should be used. By omitting the stent insertion in the absence of these criteria, operating time, postoperative pain, and analgesic requirement were reduced without increasing the complication rate. Ureteral stricturing was absent despite the low stenting rate. PMID- 11030537 TI - Ureteroscopy for proximal ureteral calculi: prevention of stone migration. AB - BACKGROUND: The cephalad migration of proximal ureteral calculi accounts for a high percentage of ureteroscopic failures. PATIENTS AND METHODS: A balloon on a wire is a single-channel 0.038-inch hollow guidewire with a low-profile balloon, which is inflatable to 12F. Forty-two consecutive patients with proximal ureteral calculi underwent ureteroscopy with the aid of this device. RESULTS: Twenty-nine patients had successful placement of the balloon on a wire above the stone. Of those 29 patients, 26 became stone free with a single procedure. CONCLUSION: The balloon on a wire is a useful tool to aid in the prevention of proximal ureteral stone migration during ureteroscopy and to minimize the number of secondary procedures. In addition, it appears to be cost-effective. PMID- 11030538 TI - Retroperitoneal laparoscopic wedge resection for small renal tumor using microwave tissue coagulator. AB - A patient with a small renal tumor underwent a successful retroperitoneal laparoscopic wedge resection using a microwave tissue coagulator without renal pedicle clamping or surface cooling. There were no postoperative complications, and renal function was well preserved. This minimally invasive procedure may be useful for treating small renal tumors. PMID- 11030539 TI - Posterior retroperitoneoscopic partial adrenalectomy using ultrasonic scalpel for aldosterone-producing adenoma. AB - BACKGROUND: Posterior retroperitoneoscopic adrenalectomy has become a standard procedure, but removal of the entire gland is not necessary in all cases. PATIENTS AND METHODS: We performed 10 posterior retroperitoneoscopic partial adrenalectomies for aldosterone-producing adenoma (APA) using an ultrasonic scalpel to divide the normal adrenal gland and the adenoma. RESULTS: The mean operation time and blood loss were 154 (110-231) minutes and 11 (5-32) mL, respectively. The mean weight of removed tissue was 5.7 g (2.3-10.2 g). Subcutaneous emphysema occurred in three patients, but there were no serious operative complications. CONCLUSION: Posterior retroperitoneoscopic partial adrenalectomy using the ultrasonic scalpel may be a valuable treatment for typical solitary APA. PMID- 11030540 TI - Long-term results of permanent indwelling wallstents for benign mid-ureteral strictures. AB - BACKGROUND: Benign ureteral strictures are a potentially difficult problem that typically has been solved by open surgery. However, minimally invasive methods would be preferable. PATIENTS AND METHODS: Three patients with benign ureteral strictures were treated with endoscopic placement of self-expanding permanent indwelling stents (Wallstents). The etiologies of the obstruction were multifactorial and included retroperitoneal fibrosis, stones, traumatic ureteroscopy, ureteral ischemia, and previous open surgery. RESULTS: One patient died with a functioning stent 1 year, 7 months after placement. The other two patients are doing well 5 years, 2 months and 5 years, 11 months afterward (mean 51 months). CONCLUSION: Insertion of a permanent indwelling self-expanding ureteral stent is relatively easy, and long-term successes are documented. Placement of Wallstents for benign ureteral strictures should be considered as a treatment alternative. PMID- 11030541 TI - Endourologic management of malignant ureteral strictures. AB - PURPOSE: This retrospective analysis assessed the efficacy of balloon dilatation, endoureterotomy, percutaneous ureteroneocystostomy with stenting, and insertion of Wallstents in the management of malignant ureteral strictures with an intact or compromised vascular supply. PATIENTS AND METHODS: A series of 127 patients with ureteral strictures secondary to malignancies were assessed after at least 2 year follow-up (range 2-5 years; mean 3.5 years). Balloon dilation (antegrade approach) was applied in 46 patients, endoureterotomy with temporary stenting in 37, percutaneous ureteroneocystostomy with stenting in 34, bougie and stents in 13, and Wallstents in 31. RESULTS: Balloon dilatation was successful in only two of four malignant midureteral stenoses with intact vascular supplies and was even less successful (10%) in midureteral strictures with a compromised vascular supply. Endoureterotomy failed in all cases to prevent ureteral obstruction. Percutaneous ureteroneocystostomy achieved patency in 11 of 34 patients (33%) having a compromised ureteral vascular supply. Wallstents were successful in 18 of 31 patients (58%) with stenoses of the pelvic ureter. CONCLUSIONS: Percutaneous ureteroneocystostomy with stenting meets the requirement for palliation in patients with obstruction secondary to pelvic neoplasms. Wallstents proved to be most successful when used in the pelvic ureter. PMID- 11030542 TI - Laparoscopic extravesicular ureteral reimplantation for vesicoureteral reflux: recent technical advances. AB - Ureteral reimplantation is an effective treatment for primary vesicoureteral reflux. Recent efforts have been directed toward reducing the perioperative morbidity of open reimplantation. We have refined the technique of laparoscopic extravesical ureteral reimplantation with emphasis on minimal tissue dissection, achieving reliable detrusor closure, and downsizing ports and instruments. With our current technique, excellent results comparable to those of established open procedures are achieved, while postoperative discomfort and the recovery period are significantly reduced. The laparoscopic technique of ureteral reimplantation is described, with emphasis on key technical modifications crucial to the ease of performance and a successful outcome. PMID- 11030543 TI - Travelling of proteins through membranes: translocation into chloroplasts. AB - Most proteins involved in plastid biogenesis are encoded by the nuclear genome. They are synthesised in the cytosol and have to be transported toward and subsequently translocated into the organelle. This targeting and import process is initiated by a specific chloroplast-targeting signal. The targeting signal of the preprotein is recognised and modified by cytosolic proteins which function in transport toward the chloroplast and in maintaining the import-competent state of the preprotein. The precursor is transferred onto a multi-component complex in the outer envelope of the chloroplasts, which is formed by receptor proteins and the translocation channel. Some proteins, not containing transit sequences, are directly sorted into the outer membrane whereas the majority, containing transit sequences, will be translocated into the stroma. This involves the joint action of a protein complex in the outer envelope, one complex in the inner envelope, and soluble proteins in the intermembrane space and the stroma. The origin of this translocation complex following the endosymbiotic events is an unsolved question. Recent identification of homologous proteins to some members of this machinery in the cyanobacterium Synechocystis PCC6803 gives an initial insight into the origin of the translocation complex. PMID- 11030544 TI - The rates of deceleration of nuclear and organellar DNA syntheses differ in the progenitor cells of the apical meristems during carrot somatic embryogenesis. AB - The synthesis of DNA in nuclei and organellar nucleoids at the various stages of somatic embryogenesis in carrot (Daucus carota L. cv. Kurodagosun) was analyzed using anti-5-bromo-2'-deoxyuridine (BrdU) immunofluorescence microscopy. The active syntheses of both nuclear and organellar DNA started in the cells forming the embryo proper 3 d after the initiation of embryogenesis, but not in cells forming suspensor-like cell aggregates. In the early globular embryo, active DNA syntheses were continuously observed in the whole embryo proper, except for the progenitor cells of the root apical meristem (RAM) and shoot apical meristem (SAM). These were recognized as slowly cycling cells with a non-BrdU-labelled nucleus and strongly BrdU-labelled organellar nucleoids. At the heart- and torpedo-shaped embryo stages, both nuclear and organellar DNA syntheses were inactive in the presumptive RAM and SAM. Thus, slowing down of organellar DNA synthesis is not coupled with, but is later than, that of nuclear DNA synthesis in the progenitor cells of the embryonic RAM and SAM. These findings clearly indicate that the timing of DNA synthesis is similar in the progenitor cells of both the RAM and SAM in the early stages of somatic embryogenesis. PMID- 11030545 TI - Expression of alpha-expansin genes in young seedlings of rice (Oryza sativa L.). AB - Rice is the only cereal in which germination and coleoptile elongation occur in hypoxia or anoxia. Little is known of the molecular basis directly underlying coleoptile cell extension. In this paper, we describe the expression of alpha expansin genes in embryos during seed development and young seedlings grown under various oxygen concentrations. The genes Os-EXP2 and Os-EXP1 were predominantly expressed in the developing seeds, mainly in newly developed leaves, coleoptiles, and seminal roots. These expansins expressed in the developing seeds may give cells the potential to expand after seed imbibition begins. In coleoptiles, Os EXP4 and Os-EXP2 mRNAs were greatly induced by submergence, while they were weakly detected in aerobic or anoxic conditions. Under submerged soil conditions, the signals hybridized with probes Os-EXP4 and Os-EXP2 in coleoptiles were strongest when coleoptiles elongated in the water layer. These data show that expansin gene expression is highly correlated with coleoptile elongation in response to oxygen concentrations. The Os-EXP4 gene was also expressed in leaves, mesocotyls, and coleorhizas of young seedlings. The growth of these tissues was also correlated with the presence of expansins. Therefore, the evidence derived from this study clearly demonstrates that expansins are indispensable for the growing tissues of rice seedlings. PMID- 11030546 TI - Calcification and measurements of net proton and oxygen flux reveal subcellular domains in Acetabularia acetabulum. AB - Vegetative adults of Acetabularia acetabulum (L.) Silva were studied as a model system for subcellular patterning in plants, and a description of several phenotypic and physiological characteristics that reveal patterns of subcellular differentiation in this unicellular macroalga was undertaken. Initially, calcification patterns were studied. Under favorable conditions, the rhizoid and most of the stalk calcified. Only the apical 10-20% of the stalk and a small region adjacent to the rhizoid remained uncalcified. Calcification in algae has been reported to result from a biologically mediated local increase in alkalinity. To test this model extracellular pH and extracellular hydrogen ion gradients were examined with ion-selective, self-referencing, electrodes. In the light, A. acetabulum displayed a general pattern of extracellular alkalinity around the entire alga, although in some individuals the region near the rhizoid and the rhizoid itself displayed extracellular acidity. Acetabularia acetabulum also displayed net hydrogen ion influx at the rhizoid and the apical half of the stalk, variable flux in the lower part of the stalk, and net hydrogen ion efflux at the base of the stalk next to the rhizoid. The lack of complete correlation between external pH patterns and calcification suggests that other factors contribute to the control of calcification in this alga. To examine whether net hydrogen ion flux patterns correlated with photosynthetic or respiration patterns, oxygen flux was measured along the stalk using self-referencing O2 electrodes. Photosynthetic oxygen evolution occurred at comparable levels throughout the stalk, with less evolution in the rhizoid. Respiration mainly occurred near and in the rhizoid, with less O2 consumption occurring more apically along the stalk. Our studies of calcification patterns, net hydrogen ion flux and O2 flux revealed several overlapping patterns of subcellular differentiation in A. acetabulum. PMID- 11030547 TI - The seed coat-specific expression of a subtilisin-like gene, SCS1, from soybean. AB - A seed coat-specific gene, SCS1 (Seed Coat Subtilisin 1), from soybean, Glycine max [L.] Merill, has been identified and studied. The gene belongs to a small family of genes with sequence similarity to the subtilisins, which are serine proteases. Northern blot analysis showed that SCS1 RNA accumulates to maximal levels in seed coats at 12 days post anthesis, preceding the final stages of seed coat differentiation. The SCS1 RNA was not found in other tissues including embryos, seed pods, flowers, stems, roots or leaves. In-situ hybridization studies confirmed the temporal pattern of expression observed by Northern blot analysis and further revealed a restricted pattern of RNA accumulation in thick walled parenchyma cells of the seed coats. These cells are important in the apoplastic translocation of nutrients en route to the embryo from the vascular tissues. The tissue-specific subtilisin-like gene may be required for regulating the differentiation of the thick-walled parenchyma cells. PMID- 11030548 TI - Control of gravimorphogenesis by auxin: accumulation pattern of CS-IAA1 mRNA in cucumber seedlings grown in space and on the ground. AB - Cucumber (Cucumis sativus L.) seedlings grown in microgravity developed a peg on each side of the transition zone between hypocotyl and root, whereas seedlings grown in a horizontal position on the ground developed a peg on the concave side of the gravitropically bending transition zone. The morphological features of the space-grown seedlings were similar to those of seedlings grown in a vertical position on the ground with their radicles pointing down: both became two-pegged seedlings. Morphogenesis of cucumber seedlings is thus inhibited by gravity. Analysis by in-situ hybridization of an auxin-inducible gene, CS-IAA1, showed that its mRNA accumulated to a much greater extent on the lower side of the transition zone in the horizontally placed seedlings on the ground just prior to and during the initiation period of peg formation. On the other hand, when seedlings were grown in microgravity or in a vertical position on the ground, accumulation of CS-IAA1 mRNA occurred all around the transition zone. Accumulation of CS-IAA1 mRNA in horizontally grown seedlings appreciably decreased on the upper side of the transition zone and increased on the lower side upon gravistimulation, compared with the two-pegged seedlings. Application of IAA to seedlings in a horizontal position caused the development of a peg on each side of the transition zone, or a collar-like protuberance, depending on the concentration used. These results suggest that upon gravistimulation the auxin concentration on the upper side of the horizontally placed transition zone is reduced to a level below the threshold value necessary for peg formation. Space grown seedlings of cucumber might develop two pegs symmetrically because the auxin level in the entire transition zone is maintained above the threshold. This spaceflight experiment verified for the first time that auxin does not redistribute in microgravity. PMID- 11030549 TI - Phenylcoumaran benzylic ether reductase, a prominent poplar xylem protein, is strongly associated with phenylpropanoid biosynthesis in lignifying cells. AB - It has previously been shown (D.R. Gang et al., 1999, J Biol Chem 274: 7516-7527) that the most abundant protein in the secondary xylem of poplar (Populus trichocarpa cv. 'Trichobel') is a phenylcoumaran benzylic ether reductase (PCBER), an enzyme involved in lignan synthesis. Here, the distribution and abundance of PCBER in poplar was studied at both the RNA and protein level. The cellular expression pattern was determined by immunolocalization of greenhouse grown plants as well as of a field-grown poplar. Compared to other poplar tissues, PCBER is preferentially produced in the secondary xylem of stems and roots and is associated with the active growth period. The protein is present in all cells of the young differentiating xylem, corresponding to the zone of active phenylpropanoid metabolism and lignification. In addition, PCBER is located in young differentiating phloem fibers, in xylem ray parenchyma, and in xylem parenchyma cells at the growth-ring border. Essentially the same expression pattern was observed in poplars grown in greenhouses and in the field. The synthesis of PCBER in phenylpropanoid-synthesizing tissues was confirmed in a bending experiment. Induction of PCBER was observed in the pith of mechanically bent poplar stems, where phenylpropanoid metabolism is induced. These results indicate that the products of PCBER activity are synthesized mainly in lignifying tissues, suggesting a role in wood development. PMID- 11030550 TI - Characterization of the sink/source transition in tobacco ( Nicotiana tabacum L.) shoots in relation to nitrogen management and leaf senescence. AB - The metabolic, biochemical and molecular events occurring during tobacco (Nicotiana tabacum) leaf ageing are presented, with a particular emphasis on nitrogen metabolism. An integrated model describing the source/sink relationship existing between leaves of different developmental stages along the main plant axis is proposed. The results of our study show that a tobacco plant can be divided into two main sections with regards to sink/source relationships. Sink-to source transition occurs at a particular leaf stage in which a breakpoint corresponding to an accumulation of carbohydrates and a depletion of both organic and inorganic nitrogen is observed. The sink/source transition is also marked by the appearence of endoproteolytic activities and the induction of both cytosolic glutamine synthetase and NAD(H)-dependent glutamate dehydrogenase transcripts, proteins and activities. The role of the newly induced enzymes and the nature of the potential metabolic and developmental signals involved in the regulation of their expression during leaf senescence are discussed. PMID- 11030551 TI - Immunolocalization of glutamine synthetase in senescing tobacco (Nicotiana tabacum L.) leaves suggests that ammonia assimilation is progressively shifted to the mesophyll cytosol. AB - Glutamine synthetase (GS) catalyses the formation of glutamine (a major form of nitrogen transport in plants) in an ATP-dependent reaction using ammonium and glutamate. This enzyme is present in the plastids and/or in the cytosol depending on the plant or the organ examined. In order to understand the role of GS isoforms in the remobilization of leaf nitrogen, we studied the localization of GS isoenzymes during natural senescence of tobacco (Nicotiana tabacum L.) leaves. Parallel to the progression of leaf senescence, an increase in cytosolic GS polypeptides was detected in the mesophyll cytosol of senescing leaves while a significant decrease in GS protein content was observed in the phloem companion cells. The presence of GS polypeptides in the leaf cytosol of senescing leaves appears to be the result of an induction of the Gln1-3 gene, the transcripts of which are not detected in mature leaves but are abundant in senescing leaves. Alltogether, our results suggest that during senescence, ammonia assimilation is progressively shifted from the chloroplasts to the cytosol of leaf mesophyll cells. PMID- 11030552 TI - Inhibition of glutathione synthesis reduces chilling tolerance in maize. AB - The role of glutathione (GSH) in protecting plants from chilling injury was analyzed in seedlings of a chilling-tolerant maize (Zea mays L.) genotype using buthionine sulfoximine (BSO), a specific inhibitor of gamma-glutamylcysteine (gammaEC) synthetase, the first enzyme of GSH synthesis. At 25 degrees C, 1 mM BSO significantly increased cysteine and reduced GSH content and GSH reductase (GR: EC 1.6.4.2) activity, but interestingly affected neither fresh weight nor dry weight nor relative injury. Application of BSO up to 1 mM during chilling at 5 degrees C reduced the fresh and dry weights of shoots and roots and increased relative injury from 10 to almost 40%. Buthionine sulfoximine also induced a decrease in GR activity of 90 and 40% in roots and shoots, respectively. Addition of GSH or gammaEC together with BSO to the nutrient solution protected the seedlings from the BSO effect by increasing the levels of GSH and GR activity in roots and shoots. During chilling, the level of abscisic acid increased both in controls and BSO-treated seedlings and decreased after chilling in roots and shoots of the controls and in the roots of BSO-treated seedlings, but increased in their shoots. Taken together, our results show that BSO did not reduce chilling tolerance of the maize genotype analyzed by inhibiting abscisic acid accumulation but by establishing a low level of GSH, which also induced a decrease in GR activity. PMID- 11030553 TI - Characterisation of pea cytosolic glutathione reductase expressed in transgenic tobacco. AB - Expression in transgenic tobacco (Nicotiana tabacum L.) of a pea (Pisum sativum L.) GOR2 cDNA, encoding an isoform of glutathione reductase (GOR2), resulted in a 3- to 7-fold elevation of total foliar glutathione reductase (GR) activity. The enzyme encoded by GOR2 was confirmed to be extraplastidial in organelle fractionation studies and was considered most likely to be localised in the cytosol. A partial purification of GOR2 was achieved but a standard affinity chromatography step, using adenosine-2',5'-diphosphate-Sepharose and often employed in the purification of GR from diverse sources, was unsuccessful with this isoform. Preparative isoelectric focussing was employed as part of the purification procedure of GOR2 and a complete separation from plastidial/mitochondrial glutathione reductase (GOR1) was achieved. The isoform GOR2 was shown to have a slower migration on non-denaturing polyacrylamide gels compared with GOR1 and properties typical of GR enzymes from plant sources. PMID- 11030554 TI - Fruit-specific lectins from banana and plantain. AB - One of the predominant proteins in the pulp of ripe bananas (Musa acuminata L.) and plantains (Musa spp.) has been identified as a lectin. The banana and plantain agglutinins (called BanLec and PlanLec, respectively) were purified in reasonable quantities using a novel isolation procedure, which prevented adsorption of the lectins onto insoluble endogenous polysaccharides. Both BanLec and PlanLec are dimeric proteins composed of two identical subunits of 15 kDa. They readily agglutinate rabbit erythrocytes and exhibit specificity towards mannose. Molecular cloning revealed that BanLec has sequence similarity to previously described lectins of the family of jacalin-related lectins, and according to molecular modelling studies has the same overall fold and three dimensional structure. The identification of BanLec and PlanLec demonstrates the occurrence of jacalin-related lectins in monocot species, suggesting that these lectins are more widespread among higher plants than is actually believed. The banana and plantain lectins are also the first documented examples of jacalin related lectins, which are abundantly present in the pulp of mature fruits but are apparently absent from other tissues. However, after treatment of intact plants with methyl jasmonate, BanLec is also clearly induced in leaves. The banana lectin is a powerful murine T-cell mitogen. The relevance of the mitogenicity of the banana lectin is discussed in terms of both the physiological role of the lectin and the impact on food safety. PMID- 11030555 TI - Effects of ultraviolet radiation on photosynthesis and related enzyme reactions of marine macroalgae. AB - Changes in physiological parameters related to photosynthesis were studied in five macroalgal species from Spitsbergen (Monostroma arcticum, Laminaria solidungula, Alaria esculenta, Palmaria palmata, Phycodrys rubens) during a 72-h exposure to UV radiation. Maximal quantum yield of photochemistry (Fv/Fm) and maximal electron transport rate (ETRmax) were measured with a pulse-amplitude modulated fluorometer; the activity of the Calvin cycle enzymes ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) were estimated using a photometric test. Proteins of crude extracts were separated by SDS gel electrophoresis and changes in cellular concentrations of Rubisco were determined. Moreover, the concentration of chlorophyll a (Ch1 a), and protein content, were measured photometrically. In all species, Ch1 a content, maximal quantum yield as well as ETRmax decreased during the UV treatment. Changes in ETRmax were related to the changes in the overall activity of Rubisco. Analysis of SDS gels showed that in P. rubens, L. solidungula, M. arcticum and A. esculenta decreasing Rubisco activity partly resulted from a degradation of the enzyme. However, in A. esculenta, the formation of a high-molecular-weight polypeptide was observed. In all species, the activity of Rubisco was more strongly impaired than that of G3PDH. Exposure to UV resulted in loss of total protein only in the deepwater species L. solidungula and P. rubens. The different sensitivities to UV exposure of the species tested reflect their zonation pattern in the field. PMID- 11030556 TI - Mechanisms of primordium formation during adventitious root development from walnut cotyledon explants. AB - In walnut (Juglans regia L.), an otherwise difficult-to-root species, explants of cotyledons have been shown to generate complete roots in the absence of exogenous growth regulators. In the present study, this process of root formation was shown to follow a pattern of adventitious, rather than primary or lateral, ontogeny: (i) the arrangement of vascular bundles in the region of root formation was of the petiole type; (ii) a typical root primordium was formed at the side of the procambium within a meristematic ring of actively dividing cells located around each vascular bundle; (iii) the developing root apical meristem was connected in a lateral way with the vascular bundle of the petiole. This adventitious root formation occurred in three main stages of cell division, primordium formation and organization of apical meristem. These stages were characterized by expression of LATERAL ROOT PRIMORDIUM-1 and CHALCONE SYNTHASE genes, which were found to be sequentially expressed during the formation of the primordium. Activation of genes related to root cell differentiation started at the early stage of primordium formation prior to organization of the root apical meristem. The systematic development of adventitious root primordia at a precise site gave indications on the positional and biochemical cues that are necessary for adventitious root formation. PMID- 11030557 TI - Cytosolic heat-stress proteins Hsp17.7 class I and Hsp17.3 class II of tomato act as molecular chaperones in vivo. AB - Small heat-stress proteins (sHsps) are the most abundant stress-induced proteins with up to 20 different members in higher plants. In the cytoplasm, two different classes can be distinguished. Two cDNA clones from tomato Lycopersicon peruvianum (L.) Mill., each coding for one of the cytoplasmic sHsp subfamilies, were analyzed with respect to their transcript and protein expression, genome organization and chaperone activity. Neither type was present under control conditions but both appeared upon heat stress and in mature fruits. Expression of the class II transcript was found to be induced at slightly lower temperatures than the class I transcript. Protein analysis using class-specific antibodies revealed an identical expression pattern of both corresponding proteins. Transient expression in an Arabidopsis thaliana (L.) Heynh. cell culture showed that, despite the difference in their amino acid sequence, both classes are functionally active as chaperones in vivo, as shown by their ability to prevent thermal inactivation of firefly luciferase in a cellular environment. PMID- 11030558 TI - Expression of a heterologous expansin in transgenic tomato plants. AB - Expansins are cell wall proteins thought to play an important role in growth and other events involving cell wall modifications. Whereas the expression patterns of many isoforms have been characterised, the nature of their activity is still poorly understood. Large amounts of active expansins are necessary to undertake biochemical studies and identify their substrates. We report here the successful expression of a recombinant expansin [CsExp1, isolated from cucumber (Cucumis sativus L.) hypocotyls] in transgenic tomato (Lycopersicon esculentum Mill.) plants, under the control of a constitutive promoter. In some transformants, CsExp1 transcript and protein accumulated to high levels, and expansin activity extractable from the cell walls was increased up to about 20-fold the activity measured in wild-type plants. These results confirm the identity of the CsExp1 coding sequence, and will enable large quantities of active expansin to be obtained for further studies. PMID- 11030559 TI - Negative regulation of nitrate reductase gene expression by glutamine or asparagine accumulating in leaves of sulfur-deprived tobacco. AB - Tobacco (Nicotiana tabacum L.) plants were subjected to a prolonged period of sulfur-deprivation to characterize molecular and metabolic mechanisms that permit control of primary N-metabolism under these conditions. Prior to the appearance of chlorotic lesions, sulfur-deprived tobacco leaves showed a strong decrease in the sulfate content and changes in foliar enzyme activities, mRNA accumulation and amino-acid pools. The basic amino acids glutamine, asparagine and arginine accumulated in the leaves of sulfur-deprived plants, while the foliar concentrations of aspartate, glutamate, serine or alanine remained fairly unchanged. Maximal extractable nitrate reductase (NR; EC 1.6.6.1) activity decreased strongly in response to sulfur-deprivation. The decrease in maximal extractable NR activity was accompanied by a decline in NR transcripts while the mRNAs of the plastidic glutamine synthetase (EC 6.1.3.2) or the beta-subunit of the mitochondrial ATP synthase were much less affected. Nitrate first accumulated in leaves of tobacco during sulfur-deprivation but then declined. An appreciable amount of nitrate was, however, present in severely sulfur-depleted leaves. The repression of NR gene expression is, therefore, not related to the decrease in the leaf nitrate level. However, glutamine- and/or asparagine-mediated repression of NR gene transcription is a possible mechanism of control in situations when glutamine and asparagine accumulate in leaves and provides a feasible explanation for the reduction in NR activity during sulfur-deprivation. The removal of reduced nitrogen from primary metabolism by redirection and storage as arginine, asparagine or glutamine combined with the down-regulation of nitrate reduction via glutamine- and/or asparagine-mediated repression of NR gene transcription may contribute to maintaining a normal N/S balance during sulfur-deprivation and indicate that the co-ordination of N- and S-metabolism is retained under these conditions. PMID- 11030560 TI - Identification of high-affinity binding sites for the hepta-beta-glucoside elicitor in membranes of the model legumes Medicago truncatula and Lotus japonicus. AB - Previous studies have led to the identification and characterization of specific, high-affinity binding sites for a hepta-beta-glucoside elicitor in soybean. A survey of plant species for elicitor-binding activity reveals that among the plants tested, the hepta-beta-glucoside elicitor is only recognized by plants belonging to the legume family. We have characterized in detail the glucan elicitor-binding site in the model legume Medicago truncatula Gaertn., and partially characterized the site in Lotus japonicus. These sites have characteristics that are very similar to the one in soybean, with dissociation constants of 4.7 and 8.9 nM respectively. The elicitor-binding sites from both plants are stable during solubilization with non-ionic alkylglycoside detergents. However, differences are observed in the abundance of the binding sites and their selectivity towards structurally related analogues of the hepta-beta-glucoside elicitor. Our results suggest that similar, but perhaps not identical, binding sites for the hepta-beta-glucoside elicitor exist in diverse legumes, but not in plants outside of the legume family. PMID- 11030561 TI - A multiplex reverse transcriptase-polymerase chain reaction method for fluorescence-based semiautomated detection of gene expression in Arabidopsis thaliana. AB - A non-radioactive, rapid and sensitive method is presented for the simultaneous detection of several mRNA molecules. The technique is based on conventional first strand cDNA synthesis by reverse transcriptase, followed by multiplex polymerase chain reaction (PCR) co-amplification of several gene products in a reaction mix containing several primer sets, each including a fluorescently labeled oligonucleotide. The PCR products obtained are finally electrophoresed in a single lane of a polyacrylamide gel, in an automated DNA sequencer controlled by fragment-analysis software. The method has proven useful to efficiently detect nine mRNA transcripts, some of which are low copy number, from small specimens such as single flowers and leaves of Arabidopsis thaliana (L.) Heynh. This approach might be easily extended to other biological systems, for developmental and physiological analyses, population studies and diagnosis. PMID- 11030562 TI - Yeast secretory expression of insulin precursors. AB - Since the 1980s, recombinant human insulin for the treatment of diabetes mellitus has been produced using either the yeast Saccharomyces cerevisiae or the prokaryote Escherichia coli. Here, development of the insulin secretory expression system in S. cerevisiae and its subsequent optimisation is described. Expression of proinsulin in S. cerevisiae does not result in efficient secretion of proinsulin or insulin. However, expression of a cDNA encoding a proinsulin like molecule with deletion of threonine(B30) as a fusion protein with the S. cerevisiae alpha-factor prepro-peptide (leader), followed either by replacement of the human proinsulin C-peptide with a small C-peptide (e.g. AAK), or by direct fusion of lysine(B29) to glycine(A1), results in the efficient secretion of folded single-chain proinsulin-like molecules to the culture supernatant. The secreted single-chain insulin precursor can then be purified and subsequently converted to human insulin by tryptic transpeptidation in organic aqueous medium in the presence of a threonine ester. The leader confers secretory competence to the insulin precursor, and constructed (synthetic) leaders have been developed for efficient secretory expression of the insulin precursor in the yeasts S. cerevisiae and Pichia pastories. The Kex2 endoprotease, specific for dibasic sites, cleaves the leader-insulin precursor fusion protein in the late secretory pathway and the folded insulin precursor is secreted to the culture supernatant. However, the Kex2 endoprotease processing of the pro-peptide-insulin precursor fusion protein is incomplete and a significant part of the pro-peptide-insulin precursor fusion protein is secreted to the culture supernatant in a hyperglycosylated form. A spacer peptide localised between the leader and the insulin precursor has been developed to optimise Kex2 endoprotease processing and insulin precursor fermentation yield. PMID- 11030563 TI - Improvement of microbial strains and fermentation processes. AB - Improvement of microbial strains for the overproduction of industrial products has been the hallmark of all commercial fermentation processes. Conventionally, strain improvement has been achieved through mutation, selection, or genetic recombination. Overproduction of primary or secondary metabolites is a complex process, and successful development of improved strains requires a knowledge of physiology, pathway regulation and control, and the design of creative screening procedures. In addition, it requires mastery of the fermentation process for each new strain, as well as sound engineering know-how for mediaoptimization and the fine-tuning of process conditions. This review focuses on the various options that may be employed to improve microbial strains and addresses the complex problems of screening, the tools and technology behind the selection of targeted organisms, and the importance of process optimization. Furthermore, this review discusses new and emerging technologies and designing optimized media for tracking mutants with enhanced productivity or other desired attributes. PMID- 11030564 TI - Enslaved bacteria as new hope for plant biotechnologists. AB - The most distinguishing feature of the plant cell is a DNA-containing organelle that sets plants apart from all other organisms: the chloroplast. Compelling evidence supports an endosymbiotic origin for chloroplasts. According to this theory, chloroplasts are descendants of formerly free-living cyanobacterial ancestors which entered an endosymbiotic relationship with a pre-eukaryotic cell and were ultimately integrated into the metabolism of the host cell. Chloroplasts retain many prokaryotic features and their gene expression system still closely resembles that of their eubacterial ancestors. During the past decade, our knowledge about chloroplast biology has benefited immensely from a most remarkable methodological breakthrough: the development of transformation technologies for chloroplast genomes. Moreover, recent advances in the manipulation of higher plant chloroplast genomes have created unprecedented opportunities for the genetic engineering of plants and promise to overcome many of the problems associated with conventional transgenic technologies. This review describes the state of the art in genetic engineering of higher plant chloroplast genomes and highlights the tremendous potential of these technologies for the biotechnology of the future. PMID- 11030565 TI - Intestinal receptors for adhesive fimbriae of enterotoxigenic Escherichia coli (ETEC) K88 in swine--a review. AB - Determining the structure of the intestinal receptor for enterotoxigenic Escherichia coli (ETEC) K88 fimbriae will make it possible to develop new strategies to prevent K88+ ETEC-induced disease in pigs. Putative K88 adhesin receptors have been identified in both intestinal brush border and mucus preparations as either glycoproteins or glycolipids. Proteins with sizes of 25, 35, 40-42, 60, and 80 kDa in the intestinal mucus and 16, 23, 35, 40-70, 74, 210, and 240 kDa in brush border membranes were reported to bind specifically to K88ab and K88ac fimbriae. The factors accounting for these variable results may include the variants of K88, ages, breeds, and phenotypes of pigs, and even the sampling sites in the small intestine. Of the reported K88 receptors, only three brush border receptors, i.e., a pair of mucin-type sialoglycoproteins (210 kDa or 240 kDa), an intestinal neutral glycosphingolipid (IGLad), and a 74-kDa transferrin glycoprotein (GP74), have fulfilled the criteria as phenotype-specific K88 fimbrial receptors. Inhibiting the attachment of ETEC to intestine by modifying the receptor attachment sites has been the key for developing novel approaches to preventing ETEC-induced diarrhea in pigs. These include: (1) receptor analogs from a variety of biological sources, (2) an enteric protected protease, (3) chicken egg-yolk containing anti-K88 fimbrial antibodies, and (4) some Lactobacillus isolates producing proteinaceous components or carbohydrates interacting with mucus components. Future studies should be directed to further characterize the carbohydrate and protein moieties of receptors recognized by the K88 adhesin variants and to identify the genes responsible for susceptibility to K88+ infections. PMID- 11030566 TI - Polymer production by two newly isolated extremely halophilic archaea: application of a novel corrosion-resistant bioreactor. AB - A novel corrosion-resistant bioreactor composed of polyetherether ketone (PEEK), tech glass and silicium nitrite ceramics was constructed and applied for the cultivation of two newly isolated, extremely halophilic archaea producing poly(gamma-glutamic acid) (PGA), or poly(beta-hydroxy butyric acid) (PHB), respectively. These bacteria were isolated from hypersaline soil close to Aswan (Egypt). The isolate strain 40, which is related to the genus Natrialba, produced large amounts of PGA when cultivated on solid medium. Culture conditions were optimised applying the corrosion-resistant bioreactor. PGA production was dependent on NaCl concentration and occurred about at 20% (w/v) NaCl in the medium. A maximum cell density of about 1.6 g cell dry matter/l was obtained when the bioreactor was stirred and aerated in a batch fermentation process using proteose-peptone medium. The supernatant was monitored with respect to PGA formation, and after 90 h a maximum of 470 mg/l culture volume was detected by HPLC analysis. Culture conditions were optimized for the isolate 56, which accumulated PHB as intracellular granules. Batch fermentations in the stirred and aerated bioreactor applying acetate and n-butyric acid as carbon sources led to cell density of 2.28 g cell dry matter/l and a maximum PHB accumulation contributing to about 53% of cellular dry weight. About 4.6 g PHB were isolated from 10.6 g dried cells of strain 56, which exhibited a weight average molar mass of 2.3 x 10(5) g mol(-1) and a polydispersity of about 1.4. PMID- 11030567 TI - Growth of Photorhabdus luminescens in batch and glucose fed-batch culture. AB - Photorhabdus luminescens, a bacterial symbiont of entomopathogenic biocontrol nematodes, was grown in batch and glucose fed-batch culture. The cell density, bioluminescence, production of antibiotic substances, number of cells with inclusion bodies, glucose concentration and oxygen uptake rate were recorded. The addition of 12.4 g 1(-1) glucose prolonged the growth, and the yield almost doubled, from 6.85 g 1(-1) to 12.45 g 1(-1) dry mass. The production of antibiotic substances increased by 140%. Bioluminescence was higher in the batch culture. A shift of P. luminescens to phase II variants was not detected. PMID- 11030568 TI - 3-carbamoyl-alpha-picolinic acid production by imidase-catalyzed regioselective hydrolysis of 2,3-pyridinedicarboximide in a water-organic solvent, two-phase system. AB - 3-Carbamoyl-alpha-picolinic acid, a versatile building block for the synthesis of agrochemicals and pharmaceuticals, was prepared by imidase-catalyzed regiospecific hydrolysis of 2,3-pyridinedicarboximide with intact Arthrobacter ureafaciens O-86 cells. Reactions were carried out in a water-organic solvent, two-phase system containing cyclohexanone at low pH to avoid spontaneous random hydrolysis. Under the optimized conditions, with the periodic addition of 2,3 pyridinedicarboximide (in total, 40 mM), the 3-carbamoyl-alpha-picolinic acid yield reached 36.6 mM in the water phase, with a molar conversion yield of 91.5% and a regioisomeric purity of 94.5%, in 2 h at pH 5.5. PMID- 11030569 TI - Immobilisation of Thiobacillus ferrooxidans cells on nickel alloy fibre for ferrous sulfate oxidation. AB - The immobilisation of the iron-oxidising bacteria Thiobacillus ferrooxidans on nickel alloy fibre as support is described. This matrix showed promise for application in iron oxidation under strongly acidic conditions. The influence on the colonisation process of T. ferrooxidans exerted by the initial pH of the medium and by temperature has also been studied. Results showed that immobilisation of T. ferrooxidans cells was affected by changes of temperature between 30 degrees C and 40 degrees C and in pH from 1.4 to 2.0. PMID- 11030570 TI - Saccharide production from methanol by transposon 5 mutants derived from the extracellular polysaccharide-producing bacterium Methylobacillus sp. strain 12S. AB - A CH3OH-utilizing bacterium that has the ability to produce extracellular polysaccharide (EPS) was isolated from a soil sample, and was identified as the obligate methylotroph Methylobacillus sp. strain 12S on the basis of its 16S rDNA sequence and growth-substrate specificity. The EPS produced by strain 12S was purified and the sugar composition was analysed by GC-MS and HPLC to reveal that the EPS was a heteropolymer composed of glucosyl, galactosyl, and mannosyl residues in the molar ratio 3:1:1. In order to produce mono- and/or oligosaccharides by single-step fermentation from CH3OH, stain 12S was mutagenized by transposon 5. Among eleven EPS-deficient mutants, three strains were found to accumulate significant amounts of reducing sugars in the media. The amounts of the reducing sugars produced by the mutants ( > ca. 700 mg glucose equivalent/l) were > 11-22 times higher than those produced by the wild-type strain ( or =0.12 (X=mole fraction of DMSO in DMSO/water mixture). The result of differential scanning calorimetry indicate that the gel to liquid-crystalline phase transition temperature increased, but the LbetaI to Pbeta, phase transition temperature decreased with an increase in DMSO concentration. These results show that DMSO stabilizes the bilayer gel phase rather than the LbetaI phase at its low concentration. The solubility of phosphorylcholine, which is the same structure as the headgroup of DHPC, decreased with an increase in DMSO concentration, indicating that the interaction free energy of the hydrophilic segments of the membrane with solvents increases with an increase in DMSO concentration. On the basis of the thermodynamic analysis, the mechanism of the stabilization of the bilayer gel phase of DHPC-MLV by DMSO is discussed. The decrease in the repulsive interaction between the headgroups of the phospholipid induced by the low concentrations of DMSO in water plays an important role in this stabilization. PMID- 11030598 TI - A carboxy-terminal alpha-helical segment in the rat skeletal muscle voltage dependent Na+ channel is responsible for its interaction with the amino-terminus. AB - Cytoplasmic segments of the adult rat skeletal muscle sodium channel alpha subunit (rSkM1) comprise a major portion (approximately 40%) of the total protein and are involved in channel functions both general, such as inactivation, and isoform-specific, for example, protein kinase A modulation. Far ultraviolet circular dichroism measurements of synthetic peptides and overexpressed fusion proteins containing individual channel cytoplasmic segments suggest that cytoplasmic domains of rSkM1 contain ordered secondary structures even in the absence of adjoining transmembrane segments. Intrinsic fluorescence experiments with a nested set of carboxy-terminal deletion proteins confirm a specific interaction between the channel's amino- and carboxy-termini and identify residues 1716-1737 in the carboxy-terminus as the region that binds to the amino terminus. Circular dichroism measurements suggest that this same region is organized as an alpha-helix and that electrostatic forces may contribute to this association. The interaction of the amino- and carboxy-termini is not accompanied by secondary structure changes detectable by circular dichroism spectroscopy, but a decrease in intrinsic fluorescence indicates that this association is accompanied by a change in the environment of Trp1617. PMID- 11030599 TI - A new cationic liposome for efficient gene delivery with serum into cultured human cells: a quantitative analysis using two independent fluorescent probes. AB - Cationic liposomes are useful to transfer genes into eukaryotic cells in vitro and in vivo. However, liposomes with good transfection efficiency are often cytotoxic, and also require serum-free conditions for optimal activity. In this report, we describe a new formulation of cationic liposome containing DC-6-14, O,O'-ditetradecanoyl-N-(alpha-trimethylammonioacetyl)diethan olamine chloride, dioleoylphosphatidylethanolamine and cholesterol for gene delivery into cultured human cells. This liposome, dispersed in 5% serum-containing growth medium, efficiently delivered a plasmid DNA for GFP (green fluorescent protein) into more than 80% of the cultured human cell hybrids derived from HeLa cells and normal fibroblasts. Flow cytometric analysis revealed that the efficiency of the GFP gene expression was 40-50% in a tumor-suppressed cell hybrid, while it was greatly reduced in the tumorigenic counterpart. The enhanced GFP expression in tumor-suppressed cell hybrids was quantitatively well correlated with a prolonged presence of the plasmid DNA, which had been labeled with another fluorescent probe, ethidium monoazide, within the cells. These results suggest that a newly developed cationic liposome is useful for gene delivery in serum-containing medium into human cells and the stability of the plasmid DNA inside the cell is a crucial step in this liposome-mediated gene expression. The mechanisms by which cationic liposome mediates gene transfer into eukaryotic cells are also discussed. PMID- 11030600 TI - Flammutoxin, a cytolysin from the edible mushroom Flammulina velutipes, forms two different types of voltage-gated channels in lipid bilayer membranes. AB - Flammutoxin, a 31-kDa cardiotoxic and cytolytic protein from the edible mushroom Flammulina velutipes, has been shown to assemble into a pore-forming annular oligomer with outer and inner diameters of 10 and 5 nm on the target cells [Tomita et al., Biochem. J. 333 (1998) 129-137]. Here we studied electrophysiological properties of flammutoxin channels using planar lipid bilayer technique, and found that flammutoxin formed two types of moderately cation-selective, voltage-gated channels with smaller and larger current amplitudes (1-4.5 pA and 20-30 pA, respectively, at 20 mV) in the lipid bilayers composed of phospholipid and cholesterol. The larger-conductance single channel showed the properties of a wide water-filled pore such as a linear relationship between channel conductance and salt concentration of the bathing solution. The functional diameter of the larger-conductance channel was estimated to be 4-5 nm by measuring the current conductance in the presence of polyethylene glycols of various sizes. In contrast, the smaller-conductance single channels showed a non linear current to voltage curve and a saturating conductance to increasing salt concentration. These results suggest that the larger-conductance channel of flammutoxin corresponds to the hemolytic pore complex, while the smaller conductance channel may reflect the intermediate state(s) of the assembling toxin. PMID- 11030601 TI - Characteristics of P2X7 receptors from human B lymphocytes expressed in Xenopus oocytes. AB - Human B lymphocytes express an ATP-gated ion channel (P2Z receptor), which shares similarities with the recently identified P2X7 receptor. Using gene specific primers, we have now isolated P2X7 cDNA from the total RNA of human B lymphocytes. This hP2X7 receptor subtype was expressed in Xenopus oocytes and electrophysiologically characterized. The hP2X7 receptor is similar to, but does not completely match, P2Z of human B cells. The hP2X7 receptors resemble the P2Z receptors with regard to the ATP concentration of half maximal activation, reproducibility, permeation characteristics and lack of desensitization of the ATP-evoked currents. However, in contrast to the native lymphocytic P2Z receptor, the time course of activation of hP2X7 displayed an additional linearly increasing current component. Furthermore, a second, small and slowly deactivating current component exists only in hP2X7 expressed in oocytes. The activation and deactivation kinetics as well as permeation characteristics of hP2X7 are different from rat P2X7 recently expressed in oocytes. Unlike in mammalian cells, hP2X7 expressed in Xenopus oocytes is not sufficient to induce large non-selective pores. PMID- 11030602 TI - Early events in acute pancreatitis. AB - Studies using experimental models indicate that the earliest changes in acute pancreatitis involve intra-acinar cell events including the co-localization of lysosomal hydrolases with digestive enzyme zymogens. This co-localization phenomenon leads to trypsinogen activation and subsequent cell injury. Following acinar cell injury, a series of extra-acinar cell changes determine the severity of pancreatitis by promoting or reducing the inflammatory response and by influencing cell death events. Most patients with pancreatitis are diagnosed when acinar cell injury has already occurred. Therapies designed to modify the subsequent extra-acinar cell inflammatory process may prove useful in the management of clinical pancreatitis. PMID- 11030603 TI - Mechanisms of intracellular zymogen activation. AB - The pancreatic acinar cell is potentially the initial site of injury that begins the series of events leading to acute pancreatitis. Pathological intrapancreatic zymogen activation occurs in experimental pancreatitis in animals and in human pancreatitis. Intracellular activation has been clearly linked to aberrant zymogen processing in one form of hereditary pancreatitis; in this genetic disease a mutation in cationic trypsinogen may eliminate the degradation of any trypsin activated in the acinar cell. Recent studies have also provided the first direct evidence that trypsinogen activation takes place early in the course of caerulein-induced pancreatitis; parallel studies have used isolated pancreatic acini and conditions that simulate those that cause pancreatitis in vivo to demonstrate that zymogens can be pathologically activated in isolated cells. A unique acinar cell pathway regulates the intracellular proteinase processing of zymogens to their active forms. Stimulating the acinar cell with supramaximal concentrations of cholecystokinin (CCK) or carbamylcholine can activate this pathway. The activation depends on a low pH compartment within the acinar cell and activation of an intracellular serine protease. A marker of trypsinogen processing, the trypsinogen activation peptide (TAP), is generated in acinar cell compartments that do not overlap with secretory granules. This compartment overlaps with a marker of recycling endosomes and lysosomes. Thus, zymogen processing within the acinar cell proceeds in a distinct subcellular compartment and is dependent on a low pH environment and activation of serine proteases. PMID- 11030604 TI - Intracellular free ionized calcium in the pathogenesis of acute pancreatitis. AB - Acute pancreatitis is a common, often severe disease with multiple causes. Many of the aetiological factors responsible for triggering acute pancreatitis have been identified but the pathophysiological mechanism by which they do so is still poorly understood. Free calcium ions within the cytosol of the acinar cell ([Ca2+]i) act as a key intracellular second messenger in the processes of stimulus-secretion coupling and may be crucial in the pathogenesis of acute pancreatitis. [Ca2+]i signals have been shown to be disrupted early in experimental pancreatitis, and it is known that an abnormal rise in [Ca2+]i is toxic by a variety of mechanisms. It has been demonstrated that abnormal, prolonged elevations in [Ca2+]i result from caerulein hyperstimulation and ethanol treatment, and it is likely that all the known causes of acute pancreatitis can cause similar disruptions. Elevations in [Ca2+]i have also been shown to be associated with both acinar cell vacuolization and intracellular enzyme activation, both of which are key steps in the pathogenesis of acute pancreatitis. A disturbance of intracellular Ca2+ signalling and the generation of an abnormal elevation in [Ca2+]i appears to be the common factor linking all the known triggers for acute pancreatitis and initiating the further sequence of pathological events leading to clinical disease. PMID- 11030605 TI - Hereditary pancreatitis: new insights, new directions. AB - Hereditary pancreatitis is an unusual form of acute and chronic pancreatitis with a familial predisposition. Recently, the gene mutations causing most cases of hereditary pancreatitis have been identified in the cationic trypsinogen gene. The known mutations are trypsinogen R117H and N211. These may predispose to acute pancreatitis by eliminating one of the fail-safe mechanisms used by the pancreas to eliminate prematurely activated trypsin. Accumulation of active trypsin mutants are hypothesized to initiate a digestive enzyme activation cascade in the pancreatic acinar cells leading to autodigestion, an intense inflammatory response, and acute pancreatitis. The observation that these patients also develop typical chronic pancreatitis and may later develop pancreatic cancer provides strong evidence that these conditions are linked. Knowledge of the pathophysiological conditions leading to acute and chronic pancreatitis and the development of a transgenic mouse expressing the mutant human trypsinogen genes will provide directions and tools necessary for the effective treatment or prevention of this human disease. PMID- 11030606 TI - Cytokines and acute pancreatitis. AB - Cytokines have been shown to play a pivotal role in multiple organ dysfunction, a major cause of death in severe acute pancreatitis. Moreover, the two-hit hypothesis of the cytokine-induced systemic inflammatory response syndrome explains the variable individual response to severe acute pancreatitis and the impact of secondary events such as sepsis or therapeutic intervention. Many experimental anti-cytokine therapies have been administered following induction of experimental pancreatitis, and have proved to be therapeutic. Patients with severe pancreatitis present early because of pain. Clearly then a window for therapeutic intervention is available between onset of symptoms and peak pro inflammatory cytokine expression. It is this fundamental observation that convinces many in the field that the treatment of AP will be one of the first clinical successes for novel drugs or therapy that seek to modulate the inflammatory response. PMID- 11030607 TI - Natural history of acute pancreatitis and the role of infection. AB - Bacterial infection of pancreatic necrotic tissue is a frequent complication of severe acute pancreatitis. Infected pancreatic necrotic tissue is observed in 30 70% of all patients suffering from necrotizing pancreatitis. It is the leading cause of deaths in severe acute pancreatitis, with mortality rates ranging from 15 to 30%. The incidence of infection increases with the extent of the necrotic areas and with the time after onset of pancreatitis. Compared to patients with sterile necrosis, those with infection of the necrotic areas have an increased mortality, and systemic complications occur more frequently. Standard treatment for infected pancreatic necrotic tissue is surgical debridement, whereas conservative management is feasible in approximately 30% of the patients with sterile necrosis. As bacterial infection of pancreatic necrotic tissue has a tremendous impact on the prognosis of the disease and on the patient's clinical course, efforts have been made to prevent it. Although clinical and experimental data provide evidence that prophylactic antibiotics have beneficial effects on the outcome and course of patients with severe acute pancreatitis, this topic has to be investigated further. General recommendations concerning the early use of antibiotics have to await the results of larger, double-blind studies. PMID- 11030608 TI - Changing concepts in the surgical management of acute pancreatitis. AB - Most episodes of acute pancreatitis are mild and self-limiting, but severe disease complicated by multiple system organ failure develops in up to 20% of cases. Early detection of those patients who subsequently develop necrotizing pancreatitis allows the start of supportive treatment in the intensive care unit before organ failure occurs. Conservative treatment in the intensive care unit, including the administration of intravenous antibiotics, is the gold standard. Surgery is indicated in patients with infected pancreatic necrosis but not in patients with sterile necrosis in the absence of deteriorating multi-organ failure despite maximal intensive care unit treatment, or other specific surgical complications. At our institution, out of 44 patients with necrotizing pancreatitis 29 (66%) had sterile necrosis and were managed conservatively while 15 (34%) had infected pancreatic necrosis and were treated by necrosectomy and continuous closed retroperitoneal lavage. There were two deaths resulting in an overall mortality of 5% in patients with severe acute pancreatitis. PMID- 11030609 TI - Antimicrobial prophylaxis in acute pancreatitis: selective decontamination versus antibiotics. AB - The results of several controlled clinical trials, published during the last 5 years, provide evidence of a beneficial role for early antimicrobial prophylaxis in severe acute pancreatitis. Pancreatic infections, especially gram-negative, which are of major importance with regard to morbidity and mortality, are gut derived. Early enteral administration of antibiotics therefore seems the most logical measure to nip the danger in the bud. Intravenous antibiotics should adequately penetrate (peri)pancreatic tissues, i.e. necrotic tissues, and should be effective against the prevalent flora in infected necrotic tissues. However, the optimal route of administration is still a matter of debate. In contrast to one clinical trial using selective decontamination (SD) (i.e. enteral antibiotics combined with short systemic prophylaxis until SD is established), no clinical trial using intravenous antibiotics has been reported in which both pancreatic infections as well as mortality were reduced. Although the evidence supporting enteral administration, i.e. SD, is not unimpressive, further controlled clinical trials, in which the different ways of administration are compared, are warranted. PMID- 11030610 TI - Role of ERCP and endoscopic sphincterotomy in acute pancreatitis. AB - When assessing the indications for interventional endoscopy, obstructive and non obstructive causes of acute pancreatitis should be distinguished. In non obstructive (e.g. alcoholic) pancreatitis, no data are available proving any benefit for endoscopic retrograde cholangiopancreatography (ERCP) and sphincterotomy. In obstructive (e.g. biliary) pancreatitis, the pathogenetic role of gallstones is controversial. The vast majority of gallstones initiating biliary pancreatitis pass spontaneously through the papilla of Vater into the duodenum without causing cholangitis or obstructive jaundice. Three prospective randomized published studies have attempted to answer the question of whether urgent removal of the stone improves the prognosis of patients suffering from acute pancreatitis. From these studies it can be concluded that the use of ERCP in acute biliary pancreatitis should depend on biliary symptoms: in cases of obstructive jaundice or cholangitis, bile duct stones should be removed as soon as possible; in patients without biliary complications, emergency ERCP is neither beneficial nor cost-effective; if retained stones (without biliary complications) are suspected, they can be removed electively. PMID- 11030611 TI - Enteral versus parenteral nutrition in acute pancreatitis. AB - Conventional wisdom has previously dictated that, in order to avoid stimulation of pancreatic secretion during acute pancreatitis, and thus avoid the perpetuation of the enzymatic activation from which the pancreatitis originated, enteral feeding should be avoided. With greater understanding of the potential role of the gastrointestinal tract in the development of a systemic inflammatory response within a number of scenarios, this dogma has recently been challenged. Moreover, there is some evidence to suggest that starving the gastrointestinal tract and providing nutritional support via the parenteral route may be associated with an increased incidence of septic complications. Experimental and clinical evidence suggests that feeding the gut may diminish intestinal permeability to endotoxin and diminish bacterial translocation, thus reducing the cytokine drive to the generalized inflammatory response and preventing organ dysfunction. Preliminary experience suggests that the institution of jejunal (but not gastric or duodenal) nutrition within 48 hours of the onset of severe acute pancreatitis diminishes endotoxic exposure, diminishes the cytokine and systemic inflammatory responses, avoids antioxidant consumption and does not cause the radiological appearances of the pancreas to deteriorate. These observations are paralleled by improvements in clinical outcome measures such as intensive care unit stay, septic complications and mortality. Whist parenteral nutrition continues to have a role in the management of acute pancreatitis particularly when complicated by fistulae or prolonged ileus, the early introduction of jejunal nutrition merits further investigation in acute pancreatitis. PMID- 11030612 TI - The possible role of platelet-activating factor antagonist therapy in the management of severe acute pancreatitis. AB - The purpose of this chapter is to examine the possible role of platelet activating factor (PAF) antagonist therapy as a means of modifying the systemic inflammatory response syndrome (SIRS) and multi-organ dysfunction syndrome (MODS) in the management of patients with severe acute pancreatitis (AP). Supposed specific treatments of AP have not shown clinical benefit, with antiprotease agents such as aprotinin and gabexate mesilate, as well as fresh frozen plasma, being ineffective. In addition, early peritoneal lavage, intravenous glucagon, somatostatin and octreotide have shown no benefit. PMID- 11030613 TI - Pausing for thought on the boundaries of imprinting. PMID- 11030614 TI - The ER translocon and retrotranslocation: is the shift into reverse manual or automatic? PMID- 11030615 TI - The debate about transport in the Golgi--two sides of the same coin? PMID- 11030616 TI - Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair. AB - In mammalian cells, double-strand breaks in DNA can be repaired by nonhomologous end-joining (NHEJ), a process dependent upon Ku70/80, DNA-PKcs, XRCC4, and DNA ligase IV. Starting with HeLa cell-free extracts, which promote NHEJ in a reaction dependent upon all of these proteins, we have purified a novel factor that stimulates DNA end-joining in vitro. Using a combination of phosphorus NMR, mass spectroscopy, and strong anion exchange chromatography, we identify this factor as inositol hexakisphosphate (IP6). Purified IP6 is bound by DNA-PK and specifically stimulates DNA-PK-dependent end-joining in vitro. The involvement of inositol phosphate in DNA-PK-dependent NHEJ is of particular interest since the catalytic domain of DNA-PKcs is similar to that found in the phosphatidylinositol 3 (PI 3)-kinase family. PMID- 11030617 TI - Targeted disruption of the nuclear receptor FXR/BAR impairs bile acid and lipid homeostasis. AB - Mice lacking the nuclear bile acid receptor FXR/BAR developed normally and were outwardly identical to wild-type littermates. FXR/BAR null mice were distinguished from wild-type mice by elevated serum bile acid, cholesterol, and triglycerides, increased hepatic cholesterol and triglycerides, and a proatherogenic serum lipoprotein profile. FXR/BAR null mice also had reduced bile acid pools and reduced fecal bile acid excretion due to decreased expression of the major hepatic canalicular bile acid transport protein. Bile acid repression and induction of cholesterol 7alpha-hydroxylase and the ileal bile acid binding protein, respectively, did not occur in FXR/BAR null mice, establishing the regulatory role of FXR/BAR for the expression of these genes in vivo. These data demonstrate that FXR/BAR is critical for bile acid and lipid homeostasis by virtue of its role as an intracellular bile acid sensor. PMID- 11030618 TI - swi1 and swi3 perform imprinting, pausing, and termination of DNA replication in S. pombe. AB - The developmental program of cell-type switching of S. pombe requires a strand specific imprinting event at the mating-type locus (mat1). Imprinting occurs only when mat1 is replicated in a specific direction and requires several trans-acting factors. This work shows (1) that the factors swi1p and swi3p act by pausing the replication fork at the imprinting site; and (2) that swi1p and swi3p are involved in termination at the mat1-proximal polar-terminator of replication (RTS1). A genetic screen to identify termination factors identified an allele that separated pausing/imprinting and termination functions of swip. These results suggest that swi1p and swi3p promote imprinting in novel ways both by pausing replication at mat1 and by terminating replication at RTS1. PMID- 11030619 TI - Combinatorial roles of the nuclear receptor corepressor in transcription and development. AB - Transcriptional repression plays crucial roles in diverse aspects of metazoan development, implying critical regulatory roles for corepressors such as N-CoR and SMRT. Altered patterns of transcription in tissues and cells derived from N CoR gene-deleted mice and the resulting block at specific points in CNS, erythrocyte, and thymocyte development indicated that N-CoR was a required component of short-term active repression by nuclear receptors and MAD and of a subset of long-term repression events mediated by REST/NRSF. Unexpectedly, N-CoR and a specific deacetylase were also required for transcriptional activation of one class of retinoic acid response element. Together, these findings suggest that specific combinations of corepressors and histone deacetylases mediate the gene-specific actions of DNA-bound repressors in development of multiple organ systems. PMID- 11030620 TI - Identification of a regulated pathway for nuclear pre-mRNA turnover. AB - We have identified a nuclear pathway that rapidly degrades unspliced pre-mRNAs in yeast. This involves 3'-->5' degradation by the exosome complex and 5'-->3' degradation by the exonuclease Rat1p. 3'-->5' degradation is normally the major pathway and is regulated in response to carbon source. Inhibition of pre-mRNA degradation resulted in increased levels of pre-mRNAs and spliced mRNAs. When splicing was inhibited by mutation of a splicing factor, inhibition of turnover resulted in 20- to 50-fold accumulation of pre-mRNAs, accompanied by increased mRNA production. Splicing of a reporter construct with a 3' splice site mutation was also increased on inhibition of turnover, showing competition between degradation and splicing. We propose that nuclear pre-mRNA turnover represents a novel step in the regulation of gene expression. PMID- 11030621 TI - Pax7 is required for the specification of myogenic satellite cells. AB - The paired box transcription factor Pax7 was isolated by representational difference analysis as a gene specifically expressed in cultured satellite cell derived myoblasts. In situ hybridization revealed that Pax7 was also expressed in satellite cells residing in adult muscle. Cell culture and electron microscopic analysis revealed a complete absence of satellite cells in Pax7(-/-) skeletal muscle. Surprisingly, fluorescence-activated cell sorting analysis indicated that the proportion of muscle-derived stem cells was unaffected. Importantly, stem cells from Pax7(-/-) muscle displayed almost a 10-fold increase in their ability to form hematopoietic colonies. These results demonstrate that satellite cells and muscle-derived stem cells represent distinct cell populations. Together these studies suggest that induction of Pax7 in muscle-derived stem cells induces satellite cell specification by restricting alternate developmental programs. PMID- 11030622 TI - Distinct origins of adult and embryonic blood in Xenopus. AB - Whether embryonic and adult blood derive from a single (yolk sac) or dual (yolk sac plus intraembryonic) origin is controversial. Here, we show, in Xenopus, that the yolk sac (VBI) and intraembryonic (DLP) blood compartments derive from distinct blastomeres in the 32-cell embryo. The first adult hematopoietic stem cells (HSCs) are thought to form in association with the floor of the dorsal aorta, and we have detected such aortic clusters in Xenopus using hematopoietic markers. Lineage tracing shows that the aortic clusters derive from the blastomere that gives rise to the DLP. These observations indicate that the first adult HSCs arise independently of the embryonic lineage. PMID- 11030623 TI - Ciboulot regulates actin assembly during Drosophila brain metamorphosis. AB - A dynamic actin cytoskeleton is essential for the remodeling of cell shape during development, but the specific roles of many actin partners remain unclear. Here we characterize a novel actin binding protein, Ciboulot (Cib), which plays a major role in axonal growth during Drosophila brain metamorphosis. Loss of Cib function leads to axonal growth defects in the central brain, while overexpression of the gene during development leads to overgrown projections. The Cib protein displays strong sequence similarity to beta-thymosins but has biochemical properties like profilin: the Cib-actin complex participates in actin filament assembly exclusively at the barbed end, and Cib enhances actin-based motility in vitro. Genetic experiments show that Cib and the Drosophila profilin protein Chickadee (Chic) cooperate in central brain metamorphosis. PMID- 11030624 TI - The 4 A X-ray structure of a tubulin:stathmin-like domain complex. AB - Phosphoproteins of the stathmin family interact with the alphabeta tubulin heterodimer (tubulin) and hence interfere with microtubule dynamics. The structure of the complex of GDP-tubulin with the stathmin-like domain of the neural protein RB3 reveals a head-to-tail assembly of two tubulins with a 91 residue RB3 alpha helix in which each copy of an internal duplicated sequence interacts with a different tubulin. As a result of the relative orientations adopted by tubulins and by their alpha and beta subunits, the tubulin:RB3 complex forms a curved structure. The RB3 helix thus most likely prevents incorporation of tubulin into microtubules by holding it in an assembly with a curvature very similar to that of the depolymerization products of microtubules. PMID- 11030625 TI - CENP-E as an essential component of the mitotic checkpoint in vitro. AB - Accurate chromatid separation is monitored by a checkpoint mechanism that delays anaphase onset until all centromeres are correctly attached to the mitotic spindle. Using Xenopus egg extracts, the kinetochore-associated microtubule motor protein CENP-E is now found to be required for establishing and maintaining this checkpoint. When CENP-E function is disrupted by immunodepletion or antibody addition, extracts fail to arrest in response to spindle damage. Mitotic arrest can be restored by addition of high levels of soluble MAD2, demonstrating that the absence of CENP-E eliminates kinetochore-dependent signaling but not the downstream steps in checkpoint signal transduction. Because it directly binds both to spindle microtubules and to the kinetochore-associated checkpoint kinase BUBR1, CENP-E is a central component in the vertebrate checkpoint that modulates signaling activity in a microtubule-dependent manner. PMID- 11030626 TI - The polymeric immunoglobulin receptor translocates pneumococci across human nasopharyngeal epithelial cells. AB - The polymeric immunoglobulin receptor (pIgR) plays a crucial role in mucosal immunity against microbial infection by transporting polymeric immunoglobulins (pIg) across the mucosal epithelium. We report here that the human pIgR (hpIgR) can bind to a major pneumococcal adhesin, CbpA. Expression of hpIgR in human nasopharyngeal cells and MDCK cells greatly enhanced pneumococcal adherence and invasion. The hpIgR-mediated bacterial adherence and invasion were abolished by either insertional knockout of cbpA or antibodies against either hpIgR or CbpA. In contrast, rabbit pIgR (rpIgR) did not bind to CbpA and its expression in MDCK cells did not enhance pneumococcal adherence and invasion. These results suggest that pneumococci are a novel example of a pathogen co-opting the pIg transcytosis machinery to promote translocation across a mucosal barrier. PMID- 11030627 TI - p300 interacts with the nuclear proto-oncoprotein SYT as part of the active control of cell adhesion. AB - Complexes containing p300, but not CBP, and the nuclear proto-oncoprotein SYT were detected in confluent cultures of G1-arrested cells but not in sparse cells or during S or G2. SYT sequences constitute the N-terminal segment of a fusion oncogene product, SYT-SSX, routinely detected in synovial sarcoma, an aggressive human tumor. SYT/p300 complex formation promotes cell adhesion to a fibronectin matrix, as reflected by compromise of this process in cells expressing SYT dl mutants that retain p300 binding activity and in the primary fibroblasts of p300 but not CBP heterozygous null mice. The mechanism linking the action of SYT/p300 complexes to adhesion function is, at least in part, transcription activation independent and results in proper activation of beta1 integrin, a major adhesion receptor. PMID- 11030629 TI - Fluorescence and autofluorescence. AB - Fluorescence detection is one of a series of new spectroscopic techniques currently developed for implementation in endoscopy. This technology is likely to significantly enhance our ability to detect minute lesions and to predict the histology of certain macroscopic lesions. The two fundamental approaches to the fluorescence detection of dysplasia and early malignancy are to use tissue specific endogenous (auto)fluorescence, and to furnish exogenous fluorophores that accumulate preferentially in neoplastic tissue. Tissue fluorescence can be detected by optical sampling of the mucosa using fluorescence spectroscopy or by taking the fluorescence information into an endoscopic image. The latter technique enables the rapid screening of large surface areas of mucosa. The clinical application of fluorescence detection in dysplasia and early cancer is still in its infancy, yet preliminary data already indicate that fluorescence imaging can indeed provide the endoscopist with real-time, accurate, non-invasive detection of dysplasia and early cancer. Furthermore, the feasibility to surpass the naked eye by detecting dysplastic lesions occult to standard endoscopy has already been established. PMID- 11030628 TI - p53AIP1, a potential mediator of p53-dependent apoptosis, and its regulation by Ser-46-phosphorylated p53. AB - Through direct cloning of p53 binding sequences from human genomic DNA, we have isolated a novel gene, designated p53AIP1 (p53-regulated Apoptosis-Inducing Protein 1), whose expression is inducible by wild-type p53. Ectopically expressed p53AIP1, which is localized within mitochondria, leads to apoptotic cell death through dissipation of mitochondrial A(psi)m. We have found that upon severe DNA damage, Ser-46 on p53 is phosphorylated and apoptosis is induced. In addition, substitution of Ser-46 inhibits the ability of p53 to induce apoptosis and selectively blocks expression of p53AIP1. Our results suggest that p53AIP1 is likely to play an important role in mediating p53-dependent apoptosis, and phosphorylation of Ser-46 regulates the transcriptional activation of this apoptosis-inducing gene. PMID- 11030630 TI - Chromoendoscopy and intravital staining techniques. AB - Chromoendoscopy and intravital staining techniques are synonymous methods for the endoscopic early detection of malignant changes in the intestinal tract. Endoscopic intravital staining involves the use of absorptive stains (methylene blue and Lugol's solution), contrast stains (indigo carmine) and reactive stains (Congo red). Lugol's iodine solution is used to identify superficial carcinomas in the squamous epithelium of the oesophagus. Methylene blue stains the specialized intestinal epithelium in Barrett's oesophagus and, in addition to this, is helpful in the diagnosis of dysplasia. Intravital staining with indigo carmine contributes to contrasting and accentuating changed mucosal processes. Together with Cresyl violet, contrast staining is particularly important in detecting small, early malignant changes in the colon. The use of chromoendoscopy enables a biopsy diagnosis of superficial dysplastic changes and an accurate delineation of carcinomatous areas. In conjunction with the modern video endoscopy (high-resolution endoscopy and magnification endoscopy), vital staining forms the diagnostic foundation for the detection of early malignant changes in the gastrointestinal tract. These techniques are therefore prerequisites to local endoscopic tumour therapy (mucosectomy). Despite their increasing acceptance, these methods must prove their diagnostic merit in randomized studies. PMID- 11030631 TI - Endosonography and endoscopic magnetic resonance imaging. AB - Endosonography is an important modality for the diagnosis and staging of oesophageal, gastric, colorectal and pancreatobiliary malignancy. It is also recognized as a reliable method for the evaluation of submucosal tumours of the gastrointestinal tract, for differentiating benign lesions from giant gastric folds, and for the localization of pancreatic endocrine tumours. The latest development, that of endosonographic fine-needle aspiration, provides for the cytological diagnosis of gastrointestinal tumours. This new technique may also be used for endoscopic therapy. High-frequency probes can be used to make a more accurate diagnosis of superficial carcinoma of the gastrointestinal tract, for three-dimensional imaging of gastrointestinal tumours and for intraductal ultrasonography of the bile duct and pancreatic duct. Endoscopic magnetic resonance imaging provides information not obtainable with endosonography or other modalities. It has high potential in the diagnosis and staging of gastrointestinal and pancreatobiliary tumours. PMID- 11030632 TI - Miniscopes. AB - Various miniscopes are available for cholangioscopy and pancreatoscopy, differing in the size of the external diameter and the instrumentation channel, the angulation system and the optical performance. For the per-oral route, the devices are inserted through a carrier duodenoscope. Ultra-thin instruments can be introduced into the biliopancreatic tract without the need for endoscopic sphincterotomy. Larger and steerable miniscopes are required for target biopsies and the approach to proximal lesions. Percutaneous cholangioscopy is more invasive and should be restricted to cases with a difficult anatomy or intrahepatic biliary disease. These techniques can be useful for the diagnosis of lesions that cannot be differentiated by indirect imaging methods. However, further studies are needed to evaluate their impact on clinical outcome. Controlled trials indicate that cholangioscopic lithotripsy is an effective and rapid alternative to other therapeutic methods for difficult bile duct stones. Mini-endoscopy is restricted to specific centres because of the high costs of the instruments, limited indications and the need for special endoscopic expertise. PMID- 11030633 TI - Micromachines in endoscopy. AB - Conventional endoscopy has reached a plateau in technical development, necessitating the exploration of bold new ideas in order to make further advances. One such idea is a self-navigating, independent, intelligent colonoscopic micro-robot. The design of a vehicle that can negotiate the difficult and hostile terrain of the colon is a complex task. Options include wheeled or tracked vehicles and pneumatically driven devices. The development of navigation and lesion recognition software to drive such a vehicle is also challenging. The various mathematical concepts involved in the development of such software are explored in this article. PMID- 11030634 TI - Virtual colonography. AB - Cross-sectional imaging techniques are increasingly being considered as alternative imaging modalities for colorectal screening. While CT-colonography uses air to insufflate the colon, an enema containing small amounts of paramagnetic contrast represents the basis for MR-colonography. Both techniques provide large three-dimensional data sets of the abdomen, containing the entire colon. Using sophisticated post-processing, the colon can be assessed in any desirable plane from various viewpoints. Thus even virtual endoscopic views can be rendered to assess the inside of the colon. This chapter briefly describes the technical aspects underlying both CT- and MR-colonography. Advantages and disadvantages of the two techniques are critically discussed, comparing them with one another as well as with conventional colonoscopy and barium enema. PMID- 11030635 TI - New haemostatic techniques: argon plasma coagulation. AB - Bleeding is one of the main challenges for endoscopists. Despite a large number of methods for haemostasis, several types of haemorrhage lack an adequate therapeutic remedy. Argon plasma-coagulation (APC), a new method for the non contact application of high-frequency current, promises to solve many of these problems. Among 1164 patients treated in 2349 sessions using APC, the indication was bleeding (due to tumour, intervention, angiodysplasia or coagulation disorders) in 305 patients (26%). The primary success rate was over 99%, the rebleeding rate 1.6% and the complication rate less than 1%, with zero mortality. Physical principles, indications, application parameters and results are discussed, especially in comparison with the Nd:YAG laser. APC is a new, efficacious, safe and easy-to-use method for the devitalization of tissue and haemostasis, especially in problematic cases. PMID- 11030636 TI - New haemostatic techniques: histoacryl injection, banding/endoloop ligation and haemoclipping. AB - New endoscopic modalities for the haemostasis of upper gastrointestinal bleeding include cyanoacrylate tissue glue injection for oesophageal and gastric varices, ligation using bands and loops for variceal and non-variceal bleeding, and clips for non-variceal bleeding. These new modalities aim to improve primary and secondary haemostasis rates and the safety of endoscopic treatment. Preliminary experience using these modalities has been encouraging, but prospective randomized trials using adequate patient numbers are still needed to validate their efficacy and safety. The choice of treatment will depend on the clinical context and the anatomy of the bleeding lesion. Cyanoacrylate injection, which achieves rapid haemostasis and obliteration of the treated varix, is ideally suited to acute variceal bleeding and the obliteration of large gastric varices. Bands and loops are used in conjunction with a transparent cap attachment for the elective treatment of oesophageal varices. The clip is most effective when a vessel from a non-variceal bleeding source can be identified. PMID- 11030637 TI - Endoscopic suturing. AB - Suturing at flexible endoscopy might extend the range of less invasive surgical procedures that can be performed without incision. The development of sewing machines that can place single and multiple stitches in the gastrointestinal tract is outlined. Methods of tying knots and cutting thread at flexible endoscopy have also been developed. The results of some applications, including the treatment of gastro-oesophageal reflux disease, the closure of perforations, haemostasis, and the attachment of feeding tubes and radiotelemetry capsules to tissue using endoscopic suturing, are described. PMID- 11030638 TI - Self-expandable metal stents. AB - Self-expandable metal stents are indicated for the palliation of oesophageal and biliary malignant strictures. In these indications, the main advantages of metal over plastic stents are a lower incidence of complications associated with the placement procedure (oesophageal stents) and a longer stent patency duration (biliary stents). This article describes the different models of metal stent currently available and their insertion techniques. Clinical results reported with every stent model are reviewed, particular emphasis being placed on 'global success', defined as the proportion of patients who require no endoscopic (or percutaneous) intervention after stent insertion, up to death. Recommendations for choosing between the numerous models available are inferred from this literature review and from our clinical experience. PMID- 11030639 TI - Botulinum toxin for spastic gastrointestinal disorders. AB - Botulinum toxin (BTX) is one of the most potent inhibitors of acetylcholine from nerve endings, and this accounts for its toxic properties as well as its therapeutic application in a variety of neuromuscular syndromes. This review focuses on the growing use of BTX in the so-called 'spastic' disorders of the gastrointestinal tract. These include achalasia, for which the short-term efficacy of the intrasphincteric injection of BTX has been well established. However, because of the chronicity of this condition, repeated injections of the toxin may be required at regular intervals. In contrast, the relatively short duration of action may be an advantage in disorders such as chronic anal fissure, where the benefit of this therapy has now been demonstrated in hundreds of patients. There are many other sphincteric and non-sphincteric syndromes in the gut for which the efficacy of this agent is being actively tested. These include non-cardiac chest pain, post-operative pylorospasm and sphincter of Oddi dysfunction. Skeletal muscle sphincters, such as the upper oesophageal sphincter or the external anal sphincter/puborectalis muscle, may also be targeted, with good effect. In some of these conditions, the local injection of BTX may serve as a useful therapeutic trial, facilitating the decision to institute more invasive forms of therapy. The cumulative short-term experience with BTX in the gut to date suggests that it is a relatively simple and safe therapy. The use of BTX represents a novel approach for gastrointestinal motility disorders, and the rapidly expanding list of successful applications holds promise for a more widespread use of similar agents in the future. Additional studies on long-term outcome are eagerly awaited. PMID- 11030640 TI - Endoscopic removal of submucous tumours. AB - Although gastrointestinal submucous tumours are increasingly being discovered with the development of endoscopic examination techniques, the reliable diagnosis of these lesions remains difficult at present. Before endoscopy became available, the treatment strategy for gastrointestinal submucous tumours was either surgery or observation. With the rapid evolution of therapeutic endoscopy, the endoscopic removal of various tumours in the gastrointestinal tract has been described, but the resection of submucous tumours is still reported only infrequently. Endoscopic removal of submucous tumours of the gastrointestinal tract has yet to become widespread, and this technique has not been fully evaluated. Not all patients need the treatment; however, endoscopic submucous tumour removal may be the treatment of choice for certain patient subgroups. This article reviews recent literature regarding the endoscopic removal of gastrointestinal submucous tumours and discusses the possibility of removing submucous tumours using endoscopic resection. PMID- 11030641 TI - Laparoscopic procedures. AB - Laparoscopic procedures are increasingly used in clinical surgery because of significantly faster convalescence than occurs with open surgery. These advances have been facilitated not least by modern gastroenterological procedures and the co-operation between surgeons and gastroenterologists. Laparoscopic cholecystectomy is the most evident example of such surgery. The widespread use of this minimally invasive approach was essentially made possible by the availability of endoscopic retrograde cholangiography (ERC) and ERT, since the laparoscopic removal of bile duct stones is still difficult and not always reliable. Further instances of so-called 'therapeutic splitting' are palliative procedures and the combined endoscopic/endoluminal and laparoscopic/intracavitary approach in cases of early cancer. Additionally, laparoscopic procedures are described that might be a surgical alternative to medical treatment (e.g. fundoplication and cardiomyotomy). PMID- 11030642 TI - Laparoscopic cholecystotomy. AB - Laparoscopic cholecystotomy (LCT) with primary closure of the gallbladder is a treatment option for patients with symptomatic cholecystolithiasis in a well functioning gallbladder. In contrast to other gallbladder-preserving minimally invasive or interventional methods, LCT is a one-session procedure avoiding the need for post-operative drainage of the gallbladder by a balloon catheter. LCT does not cause functional disturbances or severe bile duct injury, as is observed after laparoscopic cholecystectomy. PMID- 11030643 TI - The global carbon cycle: a test of our knowledge of earth as a system. AB - Motivated by the rapid increase in atmospheric CO2 due to human activities since the Industrial Revolution, several international scientific research programs have analyzed the role of individual components of the Earth system in the global carbon cycle. Our knowledge of the carbon cycle within the oceans, terrestrial ecosystems, and the atmosphere is sufficiently extensive to permit us to conclude that although natural processes can potentially slow the rate of increase in atmospheric CO2, there is no natural "savior" waiting to assimilate all the anthropogenically produced CO2 in the coming century. Our knowledge is insufficient to describe the interactions between the components of the Earth system and the relationship between the carbon cycle and other biogeochemical and climatological processes. Overcoming this limitation requires a systems approach. PMID- 11030644 TI - Blue-fluorescent antibodies. AB - The forte of catalytic antibodies has resided in the control of the ground-state reaction coordinate. A principle and method are now described in which antibodies can direct the outcome of photophysical and photochemical events that take place on excited-state potential energy surfaces. The key component is a chemically reactive optical sensor that provides a direct report of the dynamic interplay between protein and ligand at the active site. To illustrate the concept, we used a trans-stilbene hapten to elicit a panel of monoclonal antibodies that displayed a range of fluorescent spectral behavior when bound to a trans-stilbene substrate. Several antibodies yielded a blue fluorescence indicative of an excited-state complex or "exciplex" between trans-stilbene and the antibody. The antibodies controlled the isomerization coordinate of trans-stilbene and dynamically coupled this manifold with an active-site residue. A step was taken toward the use of antibody-based photochemical sensors for diagnostic and clinical applications. PMID- 11030645 TI - Optical gain and stimulated emission in nanocrystal quantum dots. AB - The development of optical gain in chemically synthesized semiconductor nanoparticles (nanocrystal quantum dots) has been intensely studied as the first step toward nanocrystal quantum dot lasers. We examined the competing dynamical processes involved in optical amplification and lasing in nanocrystal quantum dots and found that, despite a highly efficient intrinsic nonradiative Auger recombination, large optical gain can be developed at the wavelength of the emitting transition for close-packed solids of these dots. Narrowband stimulated emission with a pronounced gain threshold at wavelengths tunable with the size of the nanocrystal was observed, as expected from quantum confinement effects. These results unambiguously demonstrate the feasibility of nanocrystal quantum dot lasers. PMID- 11030646 TI - Graphite polyhedral crystals. AB - Polyhedral nano- and microstructures with shapes of faceted needles, rods, rings, barrels, and double-tipped pyramids, which we call graphite polyhedral crystals (GPCs), have been discovered. They were found in pores of glassy carbon. They have nanotube cores and graphite faces, and they can exhibit unusual sevenfold, ninefold, or more complex axial symmetry. Although some are giant radially extended nanotubes, Raman spectroscopy and transmission electron microscopy suggest GPCs have a degree of perfection higher than in multiwall nanotubes of similar size. The crystals are up to 1 micrometer in cross section and 5 micrometers in length, and they can probably be grown in much larger sizes. Preliminary results suggest a high electrical conductivity, strength, and chemical stability of GPC. PMID- 11030647 TI - The fall, recovery, orbit, and composition of the Tagish Lake meteorite: a new type of carbonaceous chondrite. AB - The preatmospheric mass of the Tagish Lake meteoroid was about 200,000 kilograms. Its calculated orbit indicates affinity to the Apollo asteroids with a semimajor axis in the middle of the asteroid belt, consistent with a linkage to low-albedo C, D, and P type asteroids. The mineralogy, oxygen isotope, and bulk chemical composition of recovered samples of the Tagish Lake meteorite are intermediate between CM and CI meteorites. These data suggest that the Tagish Lake meteorite may be one of the most primitive solar system materials yet studied. PMID- 11030648 TI - The last glacial-Holocene transition in southern Chile. AB - Warming at the last glacial termination in the North Atlantic region was interrupted by a period of renewed glacial activity during the Younger Dryas chronozone (YDC). The underlying mechanism of this cooling remains elusive, but hypotheses turn on whether it was a global or a North Atlantic phenomenon. Chronological, sedimentological, and palaeoecological records from sediments of small lakes in oceanic southern Chile demonstrate that there was no YDC cooling in southern Chile. It is therefore likely that there was little or no cooling in southern Pacific surface waters and hence that YDC cooling in the North Atlantic was a regional, rather than global, phenomenon. PMID- 11030649 TI - A niche maintaining germ line stem cells in the Drosophila ovary. AB - Stromal cells are thought to generate specific regulatory microenviroments or "niches" that control stem cell behavior. Characterizing stem cell niches in vivo remains an important goal that has been difficult to achieve. The individual ovarioles of the Drosophila ovary each contain about two germ line stem cells that maintain oocyte production. Here we show that anterior ovariolar somatic cells comprising three cell types act as a germ line stem cell niche. Germ line stem cells lost by normal or induced differentiation are efficiently replaced, and the ability to repopulate the niche increases the functional lifetime of ovarioles in vivo. Our studies implicate one of the somatic cell types, the cap cells, as a key niche component. PMID- 11030650 TI - Deleterious mutations and the evolution of sex. AB - It has been suggested that sexual reproduction is maintained because it reduces the load imposed by recurrent deleterious mutations. If rates of deleterious mutation per diploid genome per generation (U) exceed 1, and mutations interact synergistically, then sexuals can overcome their inherent twofold disadvantage. We have tested this hypothesis by estimating genomic point mutation rates for protein-coding genes in a range of animal taxa. We find a positive linear relationship between U and generation time. In species with short generation times, U is predicted to be far below 1, suggesting that sex is not maintained by its capacity to purge the genome of deleterious mutations. PMID- 11030651 TI - Localized Rac activation dynamics visualized in living cells. AB - Signaling proteins are thought to be tightly regulated spatially and temporally in order to generate specific and localized effects. For Rac and other small guanosine triphosphatases, binding to guanosine triphosphate leads to interaction with downstream targets and regulates subcellular localization. A method called FLAIR (fluorescence activation indicator for Rho proteins) was developed to quantify the spatio-temporal dynamics of the Rac1 nucleotide state in living cells. FLAIR revealed precise spatial control of growth factor-induced Rac activation, in membrane ruffles and in a gradient of activation at the leading edge of motile cells. FLAIR exemplifies a generally applicable approach for examining spatio-temporal control of protein activity. PMID- 11030652 TI - A myosin I isoform in the nucleus. AB - A nuclear isoform of myosin I beta that contains a unique 16-amino acid amino terminal extension has been identified. An affinity-purified antibody to the 16 amino acid peptide demonstrated nuclear staining. Confocal and electron microscopy revealed that nuclear myosin I beta colocalized with RNA polymerase II in an alpha-amanitin- and actinomycin D-sensitive manner. The antibody coimmunoprecipitated RNA polymerase II and blocked in vitro RNA synthesis. This isoform of myosin I beta appears to be in a complex with RNA polymerase II and may affect transcription. PMID- 11030653 TI - Plasma membrane compartmentalization in yeast by messenger RNA transport and a septin diffusion barrier. AB - Asymmetric localization of proteins plays a key role in many cellular processes, including cell polarity and cell fate determination. Using DNA microarray analysis, we identified a plasma membrane protein-encoding mRNA (IST2) that is transported to the bud tip by an actomyosin-based process. mRNA localization created a higher concentration of IST2 protein in the bud compared with that of the mother cell, and this asymmetry was maintained by a septin-mediated membrane diffusion barrier at the mother-bud neck. These results indicate that yeast creates distinct plasma membrane compartments, as has been described in neurons and epithelial cells. PMID- 11030654 TI - Molecular analysis of FRIGIDA, a major determinant of natural variation in Arabidopsis flowering time. AB - Vernalization, the acceleration of flowering by a long period of cold temperature, ensures that many plants overwinter vegetatively and flower in spring. In Arabidopsis, allelic variation at the FRIGIDA (FRI) locus is a major determinant of natural variation in flowering time. Dominant alleles of FRI confer late flowering, which is reversed to earliness by vernalization. We cloned FRI and analyzed the molecular basis of the allelic variation. Most of the early flowering ecotypes analyzed carry FRI alleles containing one of two different deletions that disrupt the open reading frame. Loss-of-function mutations at FRI have thus provided the basis for the evolution of many early-flowering ecotypes. PMID- 11030655 TI - Selfish DNA in protein-coding genes of Rickettsia. AB - Rickettsia conorii, the aetiological agent of Mediterranean spotted fever, is an intracellular bacterium transmitted by ticks. Preliminary analyses of the nearly complete genome sequence of R. conorii have revealed 44 occurrences of a previously undescribed palindromic repeat (150 base pairs long) throughout the genome. Unexpectedly, this repeat was found inserted in-frame within 19 different R. conorii open reading frames likely to encode functional proteins. We found the same repeat in proteins of other Rickettsia species. The finding of a mobile element inserted in many unrelated genes suggests the potential role of selfish DNA in the creation of new protein sequences. PMID- 11030656 TI - Replaying the game: hypnagogic images in normals and amnesics. AB - Participants playing the computer game Tetris reported intrusive, stereotypical, visual images of the game at sleep onset. Three amnesic patients with extensive bilateral medial temporal lobe damage produced similar hypnagogic reports despite being unable to recall playing the game, suggesting that such imagery may arise without important contribution from the declarative memory system. In addition, control participants reported images from previously played versions of the game, demonstrating that remote memories can influence the images from recent waking experience. PMID- 11030657 TI - A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I like protein. AB - Many bacterial pathogens encode a multisubunit toxin, termed cytolethal distending toxin (CDT), that induces cell cycle arrest, cytoplasm distention, and, eventually, chromatin fragmentation and cell death. In one such pathogen, Campylobacter jejuni, one of the subunits of this toxin, CdtB, was shown to exhibit features of type I deoxyribonucleases. Transient expression of this subunit in cultured cells caused marked chromatin disruption. Microinjection of low amounts of CdtB induced cytoplasmic distention and cell cycle arrest. CdtB mutants with substitutions in residues equivalent to those required for catalysis or magnesium binding in type I deoxyribonucleases did not cause chromatin disruption. CDT holotoxin containing these mutant forms of CdtB did not induce morphological changes or cell cycle arrest. PMID- 11030658 TI - Can we improve diagnosis of acute appendicitis? PMID- 11030659 TI - Treating children with speech and language impairments. PMID- 11030660 TI - The place of walk-in clinics in healthcare systems. PMID- 11030661 TI - The management of anogenital [correction of anal] warts. PMID- 11030662 TI - The practical management of claudication. PMID- 11030663 TI - Doctors revise declaration of Helsinki. PMID- 11030664 TI - BMJ wins an Ig Nobel prize. PMID- 11030665 TI - Warn the public of screening limitations, staff told. PMID- 11030666 TI - BMJ wins an Ig nobel prize PMID- 11030668 TI - Pregnant women with HIV should be started on antiretroviral drugs no later than 28 weeks PMID- 11030667 TI - EU to phase out tobacco advertising despite ruling. PMID- 11030669 TI - Human rights act does not affect the law on PVS. PMID- 11030670 TI - Irish kidney transplants fall by a quarter. PMID- 11030671 TI - Hungary launches its first "website hospital". PMID- 11030672 TI - Couple select healthy embryo to provide stem cells for sister. PMID- 11030673 TI - UK researchers told to maintain confidentiality. PMID- 11030674 TI - Anaesthetist faces 18 charges of serious professional misconduct. PMID- 11030675 TI - On-call time in health centres must be seen as working time. PMID- 11030676 TI - Randomised controlled trial of ultrasonography in diagnosis of acute appendicitis, incorporating the Alvarado score. AB - OBJECTIVES: To determine whether diagnosis by graded compression ultrasonography improves clinical outcomes for patients with suspected appendicitis. DESIGN: A randomised controlled trial comparing clinical diagnosis (control) with a diagnostic protocol incorporating ultrasonography and the Alvarado score (intervention group). SETTING: Single tertiary referral centre. PARTICIPANTS: 302 patients (age 5-82 years) referred to the surgical service with suspected appendicitis. 160 patients were randomised to the intervention group, of whom 129 underwent ultrasonography. Ultrasonography was omitted for patients with extreme Alvarado scores (1-3, 9, or 10) unless requested by the admitting surgical team. MAIN OUTCOME MEASURES: Time to operation, duration of hospital stay, and adverse outcomes, including non-therapeutic operations and delayed treatment in association with perforation. RESULTS: Sensitivity and specificity of ultrasonography were measured at 94. 7% and 88.9%, respectively. Patients in the intervention group who underwent therapeutic operation had a significantly shorter mean time to operation than patients in the control group (7.0 v 10.2 hours, P=0.016). There were no differences between groups in mean duration of hospital stay (53.4 v 54.5 hours, P=0.84), proportion of patients undergoing a non-therapeutic operation (9% v 11%, P=0.59) or delayed treatment in association with perforation (3% v 1%, P=0.45). CONCLUSION: Graded compression ultrasonography is an accurate procedure that leads to the prompt diagnosis and early treatment of many cases of appendicitis, although it does not prevent adverse outcomes or reduce length of hospital stay. PMID- 11030677 TI - Randomised controlled trial of community based speech and language therapy in preschool children. AB - OBJECTIVE: To compare routine speech and language therapy in preschool children with delayed speech and language against 12 months of "watchful waiting." DESIGN: Pragmatic randomised controlled trial. SETTING: 16 community clinics in Bristol. PARTICIPANTS: 159 preschool children with appreciable speech or language difficulties who fulfilled criteria for admission to speech and language therapy. MAIN OUTCOME MEASURES: Four quantitative measures of speech and language, assessed at 6 and 12 months; a binary variable indicating improvement, by 12 months, on the trial entry criterion. RESULTS: Improvement in auditory comprehension was significant in favour of therapy (adjusted difference in means 4.1, 95% confidence interval 0.5 to 7.6; P=0.025). No significant differences were observed for expressive language (1.4, -2.1 to 4.8; P=0.44); phonology error rate (-4.4, -12.0 to 3.3; P=0.26); language development (0.1, -0.4 to 0.6; P=0.73); or improvement on entry criterion (odds ratio 1.3, 0.67 to 2.4; P=0.46). At the end of the trial, 70% of all children still had substantial speech and language deficits. CONCLUSIONS: This study provides little evidence for the effectiveness of speech and language therapy compared with watchful waiting over 12 months. Providers of speech and language therapy should reconsider the appropriateness, timing, nature, and intensity of such therapy in preschool children. Continued research into more specific provision to subgroups of children is also needed to identify better treatment methods. The lack of resolution of difficulties for most of the children suggests that further research is needed to identify effective ways of helping this population of children. PMID- 11030678 TI - Severity of overdose after restriction of paracetamol availability: retrospective study. PMID- 11030680 TI - Email submissions from outside the united kingdom PMID- 11030679 TI - Drug Points: Anaphylactic-like reaction associated with oral budesonide. PMID- 11030681 TI - Walk-in primary medical care centres: lessons from Canada. PMID- 11030682 TI - Effect of needle length on incidence of local reactions to routine immunisation in infants aged 4 months: randomised controlled trial. AB - OBJECTIVE: To compare rates of local reactions associated with two needle sizes used to administer routine immunisations to infants. DESIGN: Randomised controlled trial. SETTING: Routine immunisation clinics in eight general practices in Buckinghamshire. PARTICIPANTS: Healthy infants attending for third primary immunisation due at 16 weeks of age: 119 infants were recruited, and 110 diary cards were analysed. INTERVENTIONS: Immunisation with 25 gauge, 16 mm, orange hub needle or 23 gauge, 25 mm, blue hub needle. MAIN OUTCOME MEASURES: Parental recordings of redness, swelling, and tenderness for three days after immunisation. RESULTS: Rate of redness with the longer needle was initially two thirds the rate with the smaller needle (relative risk 0.66 (95% confidence interval 0.45 to 0.99), P=0.04), and by the third day this had decreased to a seventh (relative risk 0.13 (0.03 to 0.56), P=0.0006). Rate of swelling with the longer needle was initially about a third that with the smaller needle (relative risk 0.39 (0.23 to 0.67), P=0.0002), and this difference remained for all three days. Rates of tenderness were also lower with the longer needle throughout follow up, but not significantly (relative risk 0.60 (0.29 to 1.25), P=0.17). CONCLUSIONS: Use of 25 mm needles significantly reduced rates of local reaction to routine infant immunisation. On average, for every five infants vaccinated, use of the longer needle instead of the shorter needle would prevent one infant from experiencing any local reaction. Vaccine manufacturers should review their policy of supplying the shorter needle in vaccine packs. PMID- 11030683 TI - General practitioners' attitudes and beliefs on antenatal testing for HIV: postal questionnaire survey. PMID- 11030684 TI - 10 minute consultation: menorrhagia. PMID- 11030686 TI - Waiting with time PMID- 11030687 TI - The nemesis of quackery PMID- 11030685 TI - Regular review: medical management of osteoarthritis. PMID- 11030688 TI - Lesson of the week: postoperative pressure sores after epidural anaesthesia. PMID- 11030689 TI - ABC of colorectal cancer. The role of clinical genetics in management. PMID- 11030690 TI - How can cigarette smuggling be reduced? PMID- 11030692 TI - An/atomy PMID- 11030691 TI - Disease impact number and population impact number: population perspectives to measures of risk and benefit. PMID- 11030693 TI - Priority setting in health care. Of course we should ask the tax payer. PMID- 11030694 TI - Priority setting in health care. Claims are not supported in research literature. PMID- 11030695 TI - Family histories of cancer in primary care. Referrals might be made on the basis of women's anxiety. PMID- 11030696 TI - Family histories of cancer in primary care. Nurse led clinic may provide better service than computer program. PMID- 11030697 TI - Should asymptomatic haemochromatosis be treated? Alternative strategies to appropriate diagnosis need testing. PMID- 11030698 TI - Should asymptomatic haemochromatosis be treated? Consequences of screening must be made known. PMID- 11030699 TI - Should asymptomatic haemochromatosis be treated? Update from Seamark and Hutchinson. PMID- 11030700 TI - WHO and industry partnership. Changes to paper served to stifle debate. PMID- 11030701 TI - WHO and industry partnership. A question of standards? PMID- 11030702 TI - WHO and industry partnership. Expert's opinions seemed to be disregarded. PMID- 11030703 TI - WHO and industry partnership. All commercial enterprises should be excluded from the development process. PMID- 11030705 TI - Restrictions on paracetamol have reduced amounts taken in single overdoses PMID- 11030704 TI - WHO and industry partnership. Inconsistencies need to be resolved. PMID- 11030706 TI - Walk-in primary care centres in canada show a lack of continuity of care with other primary care PMID- 11030707 TI - Longer needles produce fewer local reactions during infant immunisation PMID- 11030708 TI - General practitioners may need to be better informed before encouraging antenatal HIV testing PMID- 11030709 TI - Managers and professionals: competing ideologies PMID- 11030710 TI - Bisphenol A diglycidyl ether (BADGE) is a PPARgamma agonist in an ECV304 cell line. AB - Peroxisome proliferator activated receptors (PPAR)s are nuclear transcription factors of the steroid receptor super-family. One member, PPARgamma, a critical transcription factor in adipogenesis, is expressed in ECV304 cells, and when activated participates in the induction of cell death by apoptosis. Here we describe a clone of ECV304 cells, ECV-ACO.Luc, which stably expresses a reporter gene for PPAR activation. ECV-ACO.Luc respond to the PPARgamma agonists, 15-deoxy Delta(12,14) PGJ(2), and ciglitizone, by inducing luciferase expression. Furthermore, using ECV-ACO.Luc, we demonstrate that a newly described PPARgamma antagonist, bisphenol A diglycidyl ether (BADGE) has agonist activities. Similar to 15-deoxy-Delta(12,14) PGJ(2), BADGE induces PPARgamma activation, nuclear localization of the receptor, and induces cell death. PMID- 11030711 TI - Neuroprotection against hypoxic/hypoglycaemic injury after the insult by the group III metabotropic glutamate receptor agonist (R, S)-4 phosphonophenylglycine. AB - The role of group III metabotropic glutamate receptors (mGluR) in ischaemic neurodegeneration is still unsettled. In order to examine a possible modulatory effect of these receptors on ischaemia-induced damage we tested the novel selective agonist (R, S)-4-phosphonophenylglycine [(R,S)-PPG] after an hypoxic/hypoglycaemic insult in rat hippocampal slices. The recovery of population spike amplitudes in the CA1-region was used as parameter for neuronal viability. (R,S)-PPG significantly improved the recovery of synaptic transmission in the CA1-region even when applied only during the recovery period. The results imply that presynaptic glutamate release after an insult contributes to neurodegeneration. Since agonists of group III mGluR reduce neurotransmitter release - probably via presynaptic autoreceptors - we interpret the results obtained in our in vitro model of hypoxia/hypoglycaemia as support of the hypothesis that group III mGluR agonists might be beneficial drugs against diseases where excitotoxicity is one of the dominant pathological mechanisms. PMID- 11030712 TI - Adenosine A(1) receptor stimulation inhibits alpha(1)-adrenergic activation of the cardiac sarcolemmal Na(+)/H(+) exchanger. AB - Sarcolemmal Na(+)/H(+) exchanger (NHE) activity is increased by stimulation of G(q) protein-coupled receptors (G(q)PCRs), but the roles of other GPCRs are largely unknown. We determined the effects of N-[(1S,trans)-2 hydroxycyclopentyl]adenosine (GR79236), a selective agonist of the G(i)PCR adenosine A(1) receptor, on sarcolemmal NHE activity in adult rat ventricular myocytes (n=8-10 per group). NHE activity was indexed by the H(+) efflux rate after intracellular acidification, measured by microepifluorescence. GR79236 alone (0.01-10 microM) had no effect on NHE activity. However, co-administration of GR79236 inhibited, in a concentration-dependent manner, the stimulation of NHE activity by the alpha(1)-adrenoceptor agonist phenylephrine (10 microM). The inhibitory effect of GR79236 (10 microM) was abolished by (1) the selective A(1) antagonist 1,3-dipropyl-8-cyclopentylxanthine (0.1 microM), confirming an A(1) receptor-mediated action, and (2) pre-treatment with pertussis toxin (5 microgram ml(-1) for 60 min), indicating a G(i) protein-mediated mechanism. Our data suggest the existence of inhibitory crosstalk between the G(i)PCR adenosine A(1) receptor and the G(q)PCR alpha(1)-adrenoceptor in the regulation of sarcolemmal NHE activity. PMID- 11030713 TI - Cannabinoid receptor mediated inhibition of excitatory synaptic transmission in the rat hippocampal slice is developmentally regulated. AB - The cannabinoid (CB) receptor agonist WIN55,212-2 (500 nM) had no effect on the first of a pair of population spikes evoked in the CA1 region of hippocampal slices prepared from young adult (4 - 6 weeks old) rats, despite powerfully reducing paired-pulse depression. In contrast WIN55,212-2 caused a substantial depression of the single population spike (reduced to 43% control) and the field EPSP (reduced to 72% of control) recorded in slices prepared from neonatal (10 - 13 days old) rats. This effect was stereoselective and blocked by the CB(1) receptor antagonist AM281 (500 nM). The results indicate that activation of CB(1) receptors inhibits excitatory synaptic transmission in neonatal, but not adult rat hippocampus. This developmental regulation of CB(1) receptor mediated control of excitatory transmission may help explain some, but not all, of the previous discrepancies in the literature. PMID- 11030714 TI - Differential role of angiotensin II receptor subtypes on endothelial superoxide formation. AB - The physiological role of the angiotensin II AT2 receptor subtype is not fully characterized. We studied whether AT2 receptor could antagonize AT1 mediated superoxide formation in endothelial cells. In quiescent human umbilical vein endothelial cells (HUVEC) superoxide formation was measured after long-term incubation (6 h) with angiotensin II in the presence or absence of its receptor blocker candesartan (AT1) or PD123319 (AT2) using the cytochrome c assay. In separate experiments, the effects of AT2 mediated effects on activities of cellular phosphates including the src homology 2 domain containing phosphatases (SHP-1) was studied. The basal superoxide formation (0.19+/-0.03 nmol superoxide mg protein(-1) min(-1)) in HUVEC was increased by 37.1% after exposure to angiotensin II (100 nM,) which was due to an activation of a NAD(P)H oxidase. This was abolished by candesartan (1 microM) as well as the tyrosine kinase inhibitor genistein. In contrast, blockade of AT2 receptors by PD123319 enhanced the superoxide formation by 73.7% in intact cells. Stimulation of AT2 went along with an increased activity of tyrosine phosphatases in total cell lysates (29.8%) and, in particular, a marked stimulation of src homology 2 domain containing phosphatases (SHP-1, by 293.4%). The tyrosine phosphatase inhibitor vanadate, in turn, prevented the AT2 mediated effects on superoxide formation. The expression of both angiotensin II receptor subtypes AT1 and AT2 was confirmed by RT - PCR analysis. It is concluded that AT2 functionally antagonizes the AT1 induced endothelial superoxide formation by a pathway involving tyrosine phosphatases. PMID- 11030715 TI - Cyclic GMP-independent relaxation of rat pulmonary artery by spermine NONOate, a diazeniumdiolate nitric oxide donor. AB - In rat pulmonary artery pre-contracted with phenylephrine, the mechanisms of relaxation to the nitric oxide (NO) donor, spermine NONOate, were investigated. Responses to spermine NONOate were only partially blocked by the soluble guanylate cyclase inhibitor, ODQ (1H:-[1,2,4]Oxadiazolo-[4,3,-a]quinoxalin-1-one) at concentrations up to 30 microM. Ten microM ODQ gave maximal inhibition. Endothelium removal had no effect on the potency of spermine NONOate or its inhibition by ODQ. The protein kinase G inhibitor, Rp-8-Br-cGMPS (100 microM), caused minimal inhibition of spermine NONOate despite causing marked inhibition of glyceryl trinitrate and isosorbide dinitrate. Spermine NONOate (100 microM) caused a 35 fold increase in guanosine 3'5' cyclic monophosphate (cyclic GMP) above basal levels in pulmonary artery rings. ODQ (3 microM) abolished this cyclic GMP production but did not inhibit corresponding relaxant responses. Similar results were seen with another NONOate (MAHMA NONOate; 10 microM). ODQ resistant relaxation to spermine NONOate (i. e. relaxation seen in the presence of 10 microM ODQ) was inhibited by potassium (80 mM), charybdotoxin (300 nM), iberiotoxin (300 nM), apamin (100 nM), ouabain (1 mM) or thapsigargin (100 nM) but not by 4-aminopyridine (3 mM), glybenclamide (10 microM) or diltiazem (10 microM). Potassium, charybdotoxin, ouabain and thapsigargin also inhibited ODQ resistant relaxation to FK409 ((+/-)-E:-4-ethyl-2-[E:-hydroxyimino]-5-nitro-3 hexenamide). We conclude that, on rat pulmonary artery, spermine NONOate can produce cyclic GMP-independent relaxation that involves, at least in part, activation of Na(+)/K(+)-ATPase, sarco-endoplasmic reticulum Ca(2+)-ATPase and calcium-activated potassium channels. PMID- 11030716 TI - Characterization of the binding of [(125)I]-human prolactin releasing peptide (PrRP) to GPR10, a novel G protein coupled receptor. AB - GPR10 is a novel G-protein coupled receptor that is the human orthologue of rat Unknown Hypothalamic Receptor-1 (UHR-1). Human prolactin-releasing peptide (PrRP) has been identified as an endogenous ligand for GPR10, and occurs as 31 and 20 amino acid forms. The present study characterizes the binding of [(125)I]-PrRP-20 to HEK293 cells stably expressing GPR10 receptors. Specific binding of [(125)I] PrRP-20 was saturable, and analysis suggested evidence of both high and low affinity sites, with K:(D:) values of 0.026+/-0.006 and 0.57+/-0.14 nM respectively, and B(max) values of 3010+/-400 and 8570+/-2240 fmol mg protein(-1) respectively. Kinetic studies were unable to distinguish two sites, but single site analysis of association and dissociation data produced a K:(D:) of 0.012 nM. Competition studies revealed that human and rat PrRP-20 and PrRP-31 all display high affinity for GPR10. A range of other drugs which are known ligands at receptors which share limited homology with GPR10 were also tested. None of the drugs tested, including the RF-amide neuropeptide FF, demonstrated any affinity for GPR10. Human PrRP-20 failed to alter basal or forskolin-stimulated levels of intracellular cyclic AMP in HEK293-GPR10 cells, suggesting that GPR10 does not couple via either G(s) or G(i). Functional studies using measurements of intracellular calcium confirmed that human and rat PrRP-20 and PrRP-31 are all potent, full agonists at the GPR10 receptor. The response was blocked both by thapsigargin, indicating mobilization of intracellular Ca(2+) stores. These studies indicate that [(125)I]-PrRP-20 is a specific, high affinity radioligand for GPR10. The availability of this radioligand binding assay will be a valuable tool for the investigation of the key features involved in PrRP binding and studies on the localization and function of GPR10. PMID- 11030718 TI - Bicuculline, pentobarbital and diazepam modulate spontaneous GABA(A) channels in rat hippocampal neurons. AB - Spontaneously opening, chloride-selective channels that showed outward rectification were recorded in ripped-off patches from rat cultured hippocampal neurons and in cell-attached patches from rat hippocampal CA1 pyramidal neurons in slices. In both preparations, channels had multiple conductance states and the most common single-channel conductance varied. In the outside-out patches it ranged from 12 to 70 pS (Vp=40 mV) whereas in the cell-attached patches it ranged from 56 to 85 pS (-Vp=80 mV). Application of GABA to a patch showing spontaneous channel activity evoked a rapid, synchronous activation of channels. During prolonged exposure to either 5 or 100 microM GABA, the open probability of channels decreased. Application of GABA appeared to have no immediate effect on single-channel conductance. Exposure of the patches to 100 microM bicuculline caused a gradual decrease on the single-channel conductance of the spontaneous channels. The time for complete inhibition to take place was slower in the outside-out than in the cell-attached patches. Application of 100 microM pentobarbital or 1 microM diazepam caused 2 - 4 fold increase in the maximum channel conductance of low conductance (<40 pS) spontaneously active channels. The observation of spontaneously opening GABA(A) channels in cell-attached patches on neurons in slices suggests that they may have a role in neurons in vivo and could be an important site of action for some drugs such as benzodiazepines, barbiturates and general anaesthetics. PMID- 11030717 TI - Evidence that PAR-1 and PAR-2 mediate prostanoid-dependent contraction in isolated guinea-pig gallbladder. AB - We have investigated the ability of protease-activated receptor-1 (PAR-1), PAR-2, PAR-3 and PAR-4 agonists to induce contractile responses in isolated guinea-pig gallbladder. Thrombin, trypsin, mouse PAR-1 activating (SFLLRN-NH(2)) peptide, and mouse PAR-2 activating (SLIGRL-NH(2)) and human PAR-2 activating (SLIGKV NH(2)) peptides produced a concentration-dependent contractile response. Mouse PAR-4 activating (GYPGKF-NH(2)) peptide, the mouse PAR-1 reverse (NRLLFS-NH(2)) peptide, the mouse PAR-2 reverse (LRGILS-NH(2)) and human PAR-2 reverse (VKGILS NH(2)) peptides caused negligible contractile responses at the highest concentrations tested. An additive effect was observed following the contractile response induced by either trypsin or thrombin, with the addition of a different PAR agonist (SFLLRN-NH(2) and SLIGRL-NH(2), respectively). Desensitization to PAR 2 activating peptide attenuated the response to trypsin but failed to attenuate the response to PAR-1 agonists, and conversely desensitization to PAR-1 attenuated the response to thrombin but failed to alter contractile responses to PAR-2 agonists. The contractile responses produced by thrombin, trypsin, SFLLRN NH(2) and SLIGRL-NH(2) were markedly reduced in the presence of the cyclo oxygenase inhibitor, indomethacin, whilst the small contractile response produced by NRLLFS-NH(2) and LRGILS-NH(2) were insensitive to indomethacin. The contractile responses to thrombin, trypsin, SFLLRN-NH(2) and SLIGRL-NH(2) were unaffected by the presence of: the non-selective muscarinic antagonist, atropine; the nitric oxide synthase inhibitor, L-NAME; the sodium channel blocker, tetrodotoxin; the combination of selective tachykinin NK(1) and NK(2) receptor antagonists, (S)-1-[2-[3-(3,4-dichlorphenyl)-1 (3-isopropoxyphenylacetyl) piperidin-3-yl] ethyl]-4-phenyl-1 azaniabicyclo [2.2.2] octane chloride (SR140333) and (S)-N-methyl-N-[4-acetylamino-4-phenylpiperidino-2-(3, 4 dichlorophenyl)-butyl] benzamide (SR48968), respectively. The results indicate that PAR-1 and PAR-2 activation causes contractile responses in the guinea-pig gallbladder, an effect that is mediated principally by prostanoid release, and is independent of neural mechanisms. PMID- 11030719 TI - Evidence for VIP(1)/PACAP receptors in the afferent pathway mediating surgery induced fundic relaxation in the rat. AB - We previously reported activation of an inhibitory adrenergic and a non adrenergic non-cholinergic (NANC) pathway during abdominal surgery relaxing the rat gastric fundus. In the present study, we investigated the possible role of nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) in the NANC part of the surgery-induced fundic relaxation. The effect of the NO biosynthesis inhibitor N(G)-nitro-L-arginine (L-NOARG), the non-selective VIP receptor antagonist [D-p-Cl-Phe(6),Leu(17)]-VIP and the selective VIP(1) receptor antagonist [Acetyl-His(1),D-Phe(2),Lys(15),Arg(16), Leu(17)]-VIP was investigated on the non-adrenergic fundic relaxation induced by manipulation of the small intestine followed by resection of the caecum. Guanethidine partly reduced the manipulation-induced fundic relaxation. Addition of L-NOARG reduced this non adrenergic component, whereas the non-selective VIP receptor antagonist had no significant effect. Combination of L-NOARG and the non-selective VIP antagonist however further reduced the relaxation to manipulation. The selective VIP(1) receptor antagonist reduced the mean and maximal relaxation induced by abdominal surgery in the presence of guanethidine. When combined with L-NOARG, the relaxation of the gastric fundus was almost completely abolished. The VIP(1) receptor antagonist alone had no significant effect on the mean and maximal relaxation, but enhanced recovery of fundic tone. In conclusion, as VIP(1) receptors are not present in the rat gastric fundus, these results suggest that the NANC inhibitory pathway activated during abdominal surgery involves VIP(1) receptors, most likely in the afferent limb. The inhibitory neurotransmitters released at the level of the gastric fundus smooth muscle are NO and a substance different from VIP. PMID- 11030720 TI - Neuroprotective abilities of resveratrol and other red wine constituents against nitric oxide-related toxicity in cultured hippocampal neurons. AB - Animal and epidemiological studies suggest that polyphenol constituents of red wine possess antioxidant activities that favour protection against cardiovascular disease - the so-called. 'French paradox' - and possibly, central nervous system disorders such as Alzheimer's disease (AD) and ischaemia. In the present study, the potential of three major red wine derived-polyphenols to protect against toxicity induced by the nitric oxide free radical donors sodium nitroprusside (SNP) and 3-morpholinosydnonimine (SIN-1) was examined in cultured rat hippocampal cells. Both co- and post-treatments with either the stilbene resveratrol (5 - 25 microM) or the flavonoids quercetin (5 - 25 microM) and (+) catechin (1 - 10 microM) were capable of attenuating hippocampal cell death and intracellular reactive oxygen species accumulation produced by SNP (100 microM and 1 mM, respectively). However, among the phenolic compounds tested, only the flavonoids afforded significant protection against 5 mM SIN-1-induced toxicity. The effects of phenolic constituents were shared by Trolox (100 microM), a vitamin E analogue, but not by selective inhibitors of cyclo-oxygenases (COX) and lipoxygenases (LOX). Among the phenolic compounds tested, only quercetin (10 microM) inhibited 100 microM SNP-stimulated protein kinase C (PKC) activation, whereas none of them were able to attenuate nitrite accumulation caused by SNP (100 microM). Taken together, these data suggest that the neuroprotective abilities of quercetin, resveratrol, and (+)-catechin result from their antioxidant properties rather than their purported inhibitory effects on intracellular enzymes such as COX, LOX, or nitric oxide synthase. Quercetin, however, may also act via PKC to produce its protective effects. PMID- 11030721 TI - An orally active anti-apoptotic molecule (CGP 3466B) preserves mitochondria and enhances survival in an animal model of motoneuron disease. AB - Apoptosis and mitochondrial dysfunction are thought to be involved in the aetiology of neurodegenerative diseases. We have tested an orally active anti apoptotic molecule (CGP 3466B) that binds to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in an animal model with motoneuron degeneration, i.e. a mouse mutant with progressive motor neuronopathy (pmn). In pmn/pmn mice, CGP 3466B was administered orally (10 - 100 nmol kg(-1)) at the onset of the clinical symptoms (2 weeks after birth). CGP 3466B slowed disease progression as determined by a 57% increase in life-span, preservation of body weight and motor performance. This improvement was accompanied by a decreased loss of motoneurons and motoneuron fibres as well as an increase in retrograde transport. Electron microscopic analysis showed that CGP 3466B protects mitochondria which appear to be selectively disrupted in the motoneurons of pmn/pmn mice. The data support evaluation of CGP 3466B as a potential treatment for motor neuron disease. PMID- 11030723 TI - Delayed myocardial protection induced by endotoxin does not involve kinin B(1) receptors. AB - Endotoxin is known to confer a delayed protection against myocardial infarction. Lipopolysaccharide (LPS) treatment also induces the de novo synthesis of kinin B(1)-receptors that are not present in normal conditions. The aim of this study was to evaluate whether LPS-induced B(1)-receptors are implicated in the reduction of infarct size brought about by LPS. Rabbits were submitted to a 30 min coronary artery occlusion and 3-h reperfusion sequence. Six groups were studied: pretreated or not (control animals) with LPS (5 microgram kg(-1) i.v.) 24 h earlier and treated 15 min before and throughout ischaemia - reperfusion with either the B(1)-antagonist R-715 (1 mg kg(-1) h(-1)), the B(1)-agonist Sar [D-Phe(8)]-des-Arg(9)-bradykinin (15 microgram kg(-1) h(-1)) or vehicle (saline). Infarct size and area at risk were assessed by differential staining and planimetric analysis. The presence of B(1)-receptors in LPS-pretreated animals was confirmed by a decrease in mean arterial pressure in response to B(1) stimulation. LPS-pretreatment significantly reduced infarct size (6.4+/-1.7%, of area at risk vs 24.1+/-2.5% in control animals, P<0.05). This protection was not modified by B(1)-receptor antagonism (7.4+/-2.2%, NS) or stimulation (5.2+/-1.2%, NS). Neither antagonist nor agonist modified infarct size in control animals. In conclusion, these data suggest that LPS-induced myocardial protection in the rabbit is not related to concomitant de novo B(1)-receptor induction. PMID- 11030722 TI - Identification of a novel nicotinic binding site in mouse brain using [(125)I] epibatidine. AB - [(125)I]-Epibatidine binds to multiple nicotinic acetylcholine receptor (nAChR) subtypes with high affinity. In this study, [(125)I]-epibatidine was used to label and characterize a novel nAChR subtype found in mouse brain inferior colliculus, interpeduncular nucleus, and olfactory bulb homogenates. Binding of [(125)I]-epibatidine was saturable and apparently monophasic in each brain region (K:(D:)=71+/-12 pM mean+/-s.e.mean across regions) but inhibition of [(125)I] epibatidine binding (200 pM) by A85380, cytisine and (-)-nicotine was biphasic, indicating the presence of multiple binding sites. The sites with lower agonist affinity comprised 30.0+/-2.2, 58.6+/-0.1 and 48.7+/-3.3% of specific [(125)I] epibatidine (200 pM) binding in inferior colliculus, interpeduncular nucleus, and olfactory bulb homogenates, respectively. The affinity difference between A85380 sensitive and -resistant binding sites was particularly marked (approximately 1000 fold). Thus A85380 was used to differentiate agonist-sensitive and resistant sites. The pharmacological profiles of the A85380-resistant sites in each region were assessed with inhibition binding experiments, using 14 agonists and five antagonists. The profiles were indistinguishable across regions, implying that A85380-resistant [(125)I]-epibatidine binding sites in inferior colliculus, interpeduncular nucleus, and olfactory bulb represent a single nAChR subtype. The pharmacological profile of the A85380-resistant sites is very different from that previously reported for high affinity (-)-[(3)H]-nicotine-, [(125)I]-alpha-bungarotoxin-, or [(125)I]-alpha-conotoxin MII-binding sites, suggesting that they represent a novel nAChR population in mouse brain. PMID- 11030724 TI - Characterization of prostanoid receptors mediating contraction of the gastric fundus and ileum: studies using mice deficient in prostanoid receptors. AB - Receptors mediating prostanoid-induced contractions of longitudinal sections of gastric fundus and ileum were characterized by using tissues obtained from mice deficient in each type and subtype of prostanoid receptors. The fundus and ileum from mice deficient in either EP(3) (EP(3)(-/-) mice), EP(1) (EP(1)(-/-) mice) and FP (FP(-/-) mice) all showed decreased contraction to PGE(2) compared to the tissues from wild-type mice, whereas contraction of the fundus slightly increased in EP(4)(-/-) mice. 17-phenyl-PGE(2) also showed decreased contraction of the fundus from EP(3)(-/-), EP(1)(-/-) and FP(-/-) mice. Sulprostone showed decreased contraction of the fundus from EP(3)(-/-) and FP(-/-) mice, and decreased contraction of the ileum to this compound was seen in tissues from EP(3)(-/-), EP(1)(-/-) and FP(-/-) mice. In DP(-/-) mice, sulprostone showed increased contraction. DI-004 and AE-248 caused the small but concentration-dependent contraction of both tissues, and these contractions were abolished in tissues obtained from EP(1)(-/-) and EP(3)(-/-) mice, respectively, but not affected in other mice. Contractions of both fundus and ileum to PGF(2)alpha was absent at lower concentrations (10(-9) to 10(-7) M), and suppressed at higher concentrations (10(-6) to 10(-5) M) of the agonist in the FP(-/-) mice. Suppression of the contractions at the higher PGF(2)alpha concentrations was also seen in the fundus from EP(3)(-/-), EP(1)(-/-) and TP(-/-) mice and in the ileum from EP(3)(-/-) and TP(-/-) mice. Contraction of the fundus to PGD(2) was significantly enhanced in DP(-/-) mice, and contractions of the fundus and ileum to this PG decreased in FP(-/-) and EP(3)(-/-) mice. Contractions of both tissues to I-BOP was absent at 10(-9) to 10(-7) M and much suppressed at higher concentrations in TP(-/-) mice. Slight suppression to this agonist was also observed in the tissues from EP(3)(-/-) mice. PGI(2) induced small relaxation of both tissues from wild-type mice. These relaxation reactions were much potentiated in EP(3)(-/-) mice. On the other hand, significant contraction to PGI(2) was observed in both tissues obtained from IP(-/-) mice. These results show that contractions of the fundus and ileum induced by each prostanoid agonist are mediated by actions of this agonist on multiple types of prostanoid receptors and in some cases modified by its action on relaxant receptors. PMID- 11030725 TI - The antimalarial agent mefloquine inhibits ATP-sensitive K-channels. AB - The aim of this study was to determine whether antimalarial agents inhibit ATP sensitive potassium (K(ATP)) channels and thereby contribute to the observed side effects of these drugs. Mefloquine (10 - 100 microM), but not artenusate (100 microM), stimulated insulin release from pancreatic islets in vitro. Macroscopic K(ATP) currents were studied in inside-out patches excised from Xenopus oocytes expressing cloned K(ATP) channels. Mefloquine (IC(50) approximately 3 microM), quinine (IC(50) approximately 3 microM), and chloroquine inhibited the pancreatic beta-cell type of K(ATP) channel Kir6.2/SUR1. Artenusate (100 microM) was without effect. Mefloquine and quinine also blocked a truncated form of Kir6.2 (Kir6. 2DeltaC36) when expressed in the absence of SUR1. The extent of block was similar to that observed for Kir6.2/SUR1 currents. Our results suggest that inhibition of the beta-cell K(ATP) channel accounts for the ability of quinoline-based antimalarial drugs to stimulate insulin secretion, and thereby produce hypoglycaemia. The results also indicate that quinoline-based antimalarial agents inhibit K(ATP) channels by interaction with the Kir6.2 subunit. This subunit is common to beta-cell, neuronal, cardiac, skeletal muscle, and some smooth muscle K(ATP) channels suggesting that K(ATP) channel inhibition may contribute to the other side effects of these drugs, which include cardiac conduction abnormalities and neuropsychiatric disturbances. PMID- 11030726 TI - Pharmacological evaluation of the role of cytochrome P450 in intracellular calcium signalling in rat pancreatic acinar cells. AB - We have investigated whether the cytochrome P450 system is involved in Ca(2+) signalling in rat pancreatic acinar cells. Intracellular free [Ca(2+)] ([Ca(2+)](i)) was measured in collagenase-isolated cells using fura-2 microspectrofluorimetry and imaging. The imidazole P450 inhibitor ketoconazole (5 - 50 microM) inhibited [Ca(2+)](i) oscillations induced by cholecystokinin octapeptide (CCK). However, ketoconazole also raised baseline [Ca(2+)](i) when applied in the absence of CCK. These effects were mimicked by 5 - 50 microM SKF96365, an imidazole widely used as an inhibitor of Ca(2+) entry. The non imidazole P450 inhibitor proadifen (SKF525A) inhibited CCK-induced [Ca(2+)](i) oscillations at a concentration of 10 - 50 microM. Proadifen alone caused intracellular Ca(2+) release at 25 or 50 microM, but not at 10 microM. Octadecynoic acid and 1-aminobenzotriazole, structurally-unrelated non-imidazole P450 inhibitors, did not alter baseline [Ca(2+)](i) or CCK-evoked oscillations. We compared cumulative CCK dose-response relationship in control cells and in cells where P450 had been induced by prior injection of animals with beta naphthoflavone. Only minor differences were apparent, with induced cells showing some decrease in responsiveness at moderate and higher concentration of CCK (30 pM - 3 nM). Direct assessment of depletion-activated Ca(2+) entry showed no clear differences between control and induced cells. In conclusion, we could find no compelling evidence for a role of P450 in controlling Ca(2+) signalling generally, or Ca(2+) entry in particular, in pancreatic acinar cells. Induction of P450 is therefore probably toxic to acinar cells via a Ca(2+)-independent mechanism. PMID- 11030727 TI - Correolide, a nor-triterpenoid blocker of Shaker-type Kv1 channels elicits twitches in guinea-pig ileum by stimulating the enteric nervous system and enhancing neurotransmitter release. AB - Correolide (1 - 10 microM), a nortriterpene purified from Spachea correae and a selective blocker of Kv1 potassium channels, elicits repetitive twitching in guinea-pig ileum. This effect is not seen in guinea-pig duodenum, portal vein, urinary bladder or uterine strips, nor in rat or mouse ileum. The time course and amplitude of the correolide-induced twitches in guinea-pig ileum are similar to those elicited by electrical stimulation of the enteric nervous system. The correolide-induced twitching is not affected by pre-treatment with capsaicin (1 microM), but is facilitated by the NO synthase inhibitor, N(G)-nitro-L-arginine methyl esther (L-NAME, 200 microM). The correolide-induced twitching is abolished by tetrodotoxin (1 microM) or hexamethonium (100 microM), and is markedly inhibited by nifedipine (0.3 microM) or atropine (0.2 microM). The atropine resistant component is inhibited by selective antagonists of NK1 and NK2 tachykinin receptors, namely GR 82334 and GR 94800 (1 microM each). The former compound is more effective in inhibiting the correolide-induced, atropine resistant activity. Correolide intensified the twitching of ileum segments exposed to saturating concentrations of margatoxin (MgTX), which suggests that Kv1 sub-types other than Kv1.1 (Kv1.4 or Kv1.5) are involved in the relatively greater degree of stimulation of the enteric nervous system by correolide, as compared to MgTX. We propose that blockade of Kv1 channels by correolide increases the excitability of intramural nerve plexuses promoting release of acetylcholine and tachykinins from excitatory motor neurons. This, in turn, leads to Ca(2+)-dependent action potentials and twitching of the muscle fibres. PMID- 11030729 TI - Alpha-2 adrenoceptors are present in rat aorta smooth muscle cells, and their action is mediated by ATP-sensitive K(+) channels. AB - The role of alpha(2)-adrenoceptors in the response of aorta smooth muscle rings to the alpha(2)-adrenoceptors agonists UK 14,304 and clonidine was studied. Stimulation by 1 - 10 nM UK 14,304 caused dose-dependent relaxant responses in BaCl(2)-contracted endothelium-denuded aorta rings, and hyperpolarization in rings with or without endothelium, which were inhibited by yohimbine and glibenclamide, but not affected by prazosin, propranolol, apamin or iberiotoxin. At higher concentrations (10 nM - 10 microM) UK 14,304 also induced a depolarizing effect which was potentiated by yohimbine and inhibited by prazosin. These results indicate that UK 14,304 acts on alpha(2)-adrenoceptors at lower concentrations and on both alpha(1)- and alpha(2)-adrenoceptors above 10 nM. In rings, with or without endothelium, noradrenaline had a depolarizing effect which was inhibited by prazosin. Adrenaline did not affect the membrane potential but in the presence of prazosin caused hyperpolarization, which was inhibited by yohimbine and glibenclamide. These results indicate that noradrenaline is more selective for alpha(1)-, whereas adrenaline has similar affinities for alpha(1)- and alpha(2)-adrenoceptors. In aortae with endothelium, L-NNA caused a small depolarization but did not affect the hyperpolarization induced by UK 14,304, indicating that NO is not involved in that response. Glibenclamide induced a small depolarization in aortae, with or without endothelium, indicating that ATP sensitive K(+) channels may play a role in maintaining the smooth muscle's membrane potential. Our results indicate that, in rat aorta, alpha(2) adrenoceptors are also present in the smooth muscle, and that these receptors act through small-conductance ATP-sensitive K(+) channels. PMID- 11030728 TI - Short- and long-term differential effects of neuroprotective drug NS-7 on voltage dependent sodium channels in adrenal chromaffin cells. AB - In cultured bovine adrenal chromaffin cells, NS-7 [4-(4-fluorophenyl)-2-methyl-6 (5-piperidinopentyloxy) pyrimidine hydrochloride], a newly-synthesized neuroprotective drug, inhibited veratridine-induced (22)Na(+) influx via voltage dependent Na(+) channels (IC(50)=11.4 microM). The inhibition by NS-7 occurred in the presence of ouabain, an inhibitor of Na(+),K(+) ATPase, but disappeared at higher concentration of veratridine, and upon the washout of NS-7. NS-7 attenuated veratridine-induced (45)Ca(2+) influx via voltage-dependent Ca(2+) channels (IC(50)=20.0 microM) and catecholamine secretion (IC(50)=25.8 microM). Chronic (>/=12 h) treatment of cells with NS-7 increased cell surface [(3)H]-STX binding by 86% (EC(50)=10.5 microM; t(1/2)=27 h), but did not alter the K(D) value; it was prevented by cycloheximide, an inhibitor of protein synthesis, or brefeldin A, an inhibitor of vesicular transport from the trans-Golgi network, but was not associated with increased levels of Na(+) channel alpha- and beta(1) subunit mRNAs. In cells subjected to chronic NS-7 treatment, (22)Na(+) influx caused by veratridine (site 2 toxin), alpha-scorpion venom (site 3 toxin) or beta scorpion venom (site 4 toxin) was suppressed even after the extensive washout of NS-7, and veratridine-induced (22)Na(+) influx remained depressed even at higher concentration of veratridine; however, either alpha- or beta-scorpion venom, or Ptychodiscus brevis toxin-3 (site 5 toxin) enhanced veratridine-induced (22)Na(+) influx as in nontreated cells. These results suggest that in the acute treatment, NS-7 binds to the site 2 and reversibly inhibits Na(+) channels, thereby reducing Ca(2+) channel gating and catecholamine secretion. Chronic treatment with NS-7 up regulates cell surface Na(+) channels via translational and externalization events, but persistently inhibits Na(+) channel gating without impairing the cooperative interaction between the functional domains of Na(+) channels. PMID- 11030730 TI - Negative feedback regulation of reactive oxygen species on AT1 receptor gene expression. AB - Free radicals as well as the AT1 receptor are involved in the pathogenesis of cardiovascular disease. Both the intracellular mechanisms of AT1 receptor regulation and the effect of free radicals on AT1 receptor expression are currently unknown. This study investigates the role of free radicals in the modulation of AT1 receptor expression and in the angiotensin II-induced AT1 receptor regulation. AT1 receptor mRNA was assessed by Northern blotting and AT1 receptor density by radioligand binding assays, respectively, in vascular smooth muscle cells (VSMC). Free radical release was measured by confocal laser scanning microscopy. AT1 receptor mRNA transcription rate was determined by nuclear run-on assays and AT1 receptor mRNA half-life was measured under transcriptional blockade. Angiotensin II caused a time-dependent decrease of AT1 receptor mRNA expression in rat VSMC in culture (30+/-6% at 4 h with 100 nM angiotensin II). This was followed by a consistent decrease in AT1 receptor density. Angiotensin II caused release of reactive oxygen species in VSMC which was abolished by preincubation with 100 microM diphenylene iodonium (DPI). DPI inhibited partially the down-regulating effect of angiotensin II on the AT1 receptor. Incubation of VSMC with either hydrogen peroxide or xanthine/xanthine oxidase caused a dose dependent decrease in AT1 receptor mRNA expression which was not mediated by a decreased rate of transcription but rather through destabilization of AT1 receptor mRNA. Experiments which included preincubation of VSMC with various intracellular inhibitors suggested that free radicals caused AT1 receptor downregulation through activation of p38-MAP kinase and intracellular release of calcium. However, angiotensin II-induced AT1 receptor expression was not inhibited by blockade of p38-MAP kinase activation or intracellular calcium release. Free radicals may at least in part mediate angiotensin II-induced AT1 receptor regulation through direct post-transcriptional effects on AT1 receptor mRNA expression which involves intracellular release of calcium and activation of p38-MAP kinase. These findings may help to clarify the intracellular mechanisms involved in AT1 receptor regulation and reveal a novel biological feature for reactive oxygen species. PMID- 11030731 TI - Cyclo-oxygenase-1 and -2 contribution to endothelial dysfunction in ageing. AB - Experiments were designed to investigate the role of cyclo-oxygenase isoforms in endothelial dysfunction in ageing. Aortic rings with endothelium of aged and young (24 vs 4 month-old) Wistar rats, were mounted in organ chambers for the recording of changes in isometric tension. In young rats, acetylcholine (ACh) caused a complete relaxation which was not affected by indomethacin (0.3 microM), NS-398 (a preferential COX-2 inhibitor; 1 microM), SQ-29548 (a thromboxane receptor antagonist; 1 microM), nor valeryl-salicylate (VAS, a preferential inhibitor of COX-1; 3 mM). In aged rats, ACh caused a biphasic response characterized by a first phase of relaxation (0.01 - 1 microM ACh), followed by a contraction (3 - 100 microM ACh). Indomethacin, NS-398 and SQ-29548, but not VAS, augmented the first phase. Indomethacin, VAS, NS-398 and SQ-29548 decreased the contractions to high ACh concentrations. Then, the sensitivity to thromboxane receptor activation was investigated with U-46619. The results show comparable EC(50) values in young and aged rats. In aged rats, the ACh-stimulated release of prostacyclin, prostaglandin F(2alpha) and thromboxane A(2) was decreased by either indomethacin, NS-398, VAS or endothelium removal. However, in young animals, the ACh-stimulated release of prostacyclin and prostaglandin F(2alpha) were smaller than in older animals and remained unaffected by NS-398. Aortic endothelial cells from aged - but not young - rats express COX-2 isoform, while COX-1 labelling was observed in endothelial cells from both young and aged rats. These data demonstrate the active contribution of COX-1 and -2 in endothelial dysfunction associated with ageing. PMID- 11030732 TI - Sub-family selective actions in the ability of Erk2 MAP kinase to phosphorylate and regulate the activity of PDE4 cyclic AMP-specific phosphodiesterases. AB - Expressed in intact cells and in vitro, PDE4B and PDE4C isoenzymes of cyclic nucleotide phosphodiesterase (PDE), in common with PDE4D isoenzymes, are shown to provide substrates for C-terminal catalytic unit phosphorylation by the extracellular signal-regulated kinase Erk2 (p42(MAPK)). In contrast, PDE4A isoenzymes do not provide substrates for C-terminal catalytic unit phosphorylation by Erk2. Mutant PDE4 enzymes were generated to show that Erk2 phosphorylation occurs at a single, cognate serine residue located within the C terminal portion of the PDE4 catalytic unit. PDE4 long-form isoenzymes were markedly inhibited by Erk2 phosphorylation. The short-form PDE4B2 isoenzyme was activated by Erk2 phosphorylation. These functional changes in PDE activity were mimicked by mutation of the target serine for Erk2 phosphorylation to the negatively charged amino acid, aspartic acid. Epidermal growth factor (EGF) challenge caused diametrically opposed changes in cyclic AMP levels in COS1 cells transfected to express the long PDE4B1 isoenzyme compared to cells expressing the short PDE4B2 isoenzyme. We suggest that PDE4 enzymes may provide a pivotal point for integrating cyclic AMP and Erk signal transduction in cells with 4 genes encoding enzymes that are either insensitive to Erk2 action or may either be activated or inhibited. This indicates that PDE4 isoenzymes have distinct functional roles, giving credence to the notion that distinct therapeutic benefits may accrue using either PDE4 subfamily or isoenzyme-selective inhibitors. PMID- 11030733 TI - In vivo transfer of antisense oligonucleotide against urinary kininase blunts deoxycorticosterone acetate-salt hypertension in rats. AB - We have previously reported that the renal kallikrein-kinin system suppressed the development of deoxycorticosterone acetate (DOCA)-salt hypertension. Kinins were degraded in the kidney mainly by carboxypeptidase Y (CPY)-like kininase. Blockade of renal kinin degradation may reduce hypertension in the developmental stage. We constructed an antisense oligonucleotide against rat CPY homologue (5'-CAT-CTC TGC-TTC-CTT-GTG-TC-3', AS) and its randomized control oligonucleotide (5'-TCC-TTC CTG-CTT-GAG-TTC-CT-3', RC), and prepared an HVJ-liposome complex that prolongs and increases the effectiveness of the antisense oligonucleotide. Antisense oligonucleotide was transfected (25 nmole rat(-1), in terms of nucleotide) into the kidney from the renal artery. Blood pressure was measured through a catheter inserted into the abdominal aorta. Mean blood pressure (MBP) in DOCA-salt treated (for 2 weeks) Sprague Dawley strain rats was 130+/-3 mmHg (n=11), and was reduced significantly (P<0.05) more by AS transfection (122+/-4 mmHg, n=6) than by RC treatment (137+/-6 mmHg, n=5) 4 days after the transfection. This reduction in MBP was accompanied by increased urinary sodium excretion (AS, 8.4+/-1.5 mmole day(-1); RC, 4.6+/-0.5 mmole day(-1), P<0.05) and a reduction in urinary CPY-like kininase activity. Ebelactone B (5 mg kg(-1), twice a day, p.o.), an inhibitor for urinary CPY-like kininase, also reduced MBP and induced natriuresis to the same degree as AS. Lisinopril, an inhibitor for angiotensin converting enzyme (ACE) failed to reduce the elevated MBP. These results suggest that CPY-like kininase may have more contribution than ACE to degrade kinin in the kidney, and that knockdown of CPY-like kininase in the kidney may partly prevent rat DOCA salt hypertension. PMID- 11030734 TI - Pharmacological characterization of the human 5-HT(4(d)) receptor splice variant stably expressed in Chinese hamster ovary cells. AB - The recently identified C-terminal splice variant of the human 5-HT(4) receptor, the h5-HT(4(d)) receptor, was stably expressed in a CHO cell line at 493+/-25 fmol mg(-1) protein. We analysed its pharmacological properties by measuring binding affinities and 5-HT(4) ligand-induced cyclic AMP production. The pharmacological binding profile determined in competition studies with the specific antagonist [(3)H]-GR113808 revealed a rank order of affinity of 5-HT(4) ligands for the h5-HT(4(d)) receptor that was consistent with those previously reported for other 5-HT(4) receptor isoforms. In adenylyl cyclase functional assays, the h5-HT(4(d)) receptor displayed equipotent coupling for all 5-HT(4) agonists tested (EC(50) in the range of 1 - 6 nM). EC(50) values were lower than those previously obtained with the 5-HT(4(e)) receptor stably expressed in CHO cells indicating that the 5-HT(4(d)) receptor was more efficiently coupled to its effector than the 5-HT(4(e)) receptor isoform. Moreover, in terms of agonist efficacy (E(max)), the benzamide derivative, renzapride displayed full agonist properties at the h5-HT(4(d)) receptor (same E(max) as 5-HT) whereas it was previously shown to be a partial agonist at the h5-HT(4(e)) receptor. A constitutive activity of the h5-HT(4(d)) receptor was observed in CHO cells in the absence of any 5-HT(4) ligand. Surprisingly, two 5-HT(4) ligands, SB204070 and RS39604 which are described as highly potent antagonists in various biological models, revealed partial agonist properties at the h5-HT(4(d)) receptor. We conclude that C-terminal tails of 5-HT(4) receptor isoforms may directly influence their functional properties. PMID- 11030735 TI - Analysis of 3-morpholinosydnonimine and sodium nitroprusside effects on dopamine release in the striatum of freely moving rats: role of nitric oxide, iron and ascorbic acid. AB - The effects of intrastriatal infusion of 3-morpholinosydnonimine (SIN-1) or sodium nitroprusside (SNP) on dopamine (DA), 3-methoxytyramine (3-MT), dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), L dihydroxyphenylalanine (L-DOPA), ascorbic acid and uric acid concentrations in dialysates from the striatum of freely moving rats were evaluated using microdialysis. SIN-1 (1 mM) infusion for 180 min increased microdialysate DA and 3-MT concentrations, while L-DOPA, DOPCA+HVA, ascorbic acid and uric acid levels were unaffected. Co-infusion with ascorbic acid (0.1 mM) inhibited SIN-1-induced increases in DA and 3-MT dialysate concentration. SNP (1 mM) infusion for 180 min increased greatly the dialysate DA concentration to a peak (2950% of baseline) at the end of the infusion, while increases in 3-MT were negligible. In addition, SNP decreased ascorbic acid and L-DOPA but increased uric acid concentration in the dialysate. Co-infusion with deferoxamine (0.2 mM) inhibited the late SNP induced increase in DA dialysate concentration, but did not affect the decrease in ascorbic acid and increase uric acid dialysate concentrations. SNP (1 mM) infusion for 20 min moderately increased uric acid, DA and 3-MT, but decreased L DOPA levels in the dialysate. Ascorbic acid concentration increased at the end of SNP infusion. Co-infusion with ascorbic acid (0.1 mM) inhibited the SNP-induced increase in DA and 3-MT, but did not affect the decrease in L-DOPA and increase in uric acid dialysate concentrations. These results suggest that NO released from SIN-1 may account for the increase in the dialysate DA concentration. NO released following decomposition of SNP may account for the early increase in dialysate DA, while late changes in microdialysate composition following SNP may result from an interaction between NO and the ferrocyanide moiety of SNP. Exogenous ascorbic acid inhibits the effect of exogenous NO on DA release probably by scavenging NO, suggesting that endogenous ascorbic acid may modulate the NO control of DA release from 300 striatal dopaminergic terminals. PMID- 11030736 TI - A novel positive allosteric modulator of the GABA(A) receptor: the action of (+) ROD188. AB - (+)-ROD188 was synthesized in the search for novel ligands of the GABA binding site. It shares some structural similarity with bicuculline. (+)-ROD188 failed to displace [(3)H]-muscimol in binding studies and failed to induce channel opening in recombinant rat alpha1beta2gamma2 GABA(A) receptors functionally expressed in Xenopus oocytes. (+)-ROD188 allosterically stimulated GABA induced currents. Displacement of [(3)H]-Ro15-1788 indicated a low affinity action at the benzodiazepine binding site. In functional studies, stimulation by (+)-ROD188 was little sensitive to the presence of 1 microM of the benzodiazepine antagonist Ro 15-1788, and (+)-ROD188 also stimulated currents mediated by alpha1beta2, indicating a major mechanism of action different from that of benzodiazepines. Allosteric stimulation by (+)-ROD188 was similar in alpha1beta2N265S as in unmutated alpha1beta2, while that by loreclezole was strongly reduced. (+)-ROD188 also strongly stimulated currents elicited by either pentobarbital or 5alpha pregnan-3alpha-ol-20-one (3alpha-OH-DHP), in line with a mode of action different from that of barbiturates or neurosteroids as channel agonists. Stimulation by (+)-ROD188 was largest in alpha6beta2gamma2 (alpha6beta2gamma2>>alpha1beta2gamma2=alpha5beta2gamma2++ +>alpha2beta2ga mma2= alpha3beta2gamma2), indicating a unique subunit isoform specificity. Miniature inhibitory postsynaptic currents (mIPSC) in cultures of rat hippocampal neurons, caused by spontaneous release of GABA showed a prolonged decay time in the presence of 30 microM (+)-ROD188, indicating an enhanced synaptic inhibitory transmission. PMID- 11030737 TI - Arachidonic acid-mediated cooxidation of all-trans-retinoic acid in microsomal fractions from human liver. AB - The quantitative importance of prostaglandin H synthase (PGHS)-mediated cooxidation of all-trans-retinoic acid (ATRA) was evaluated in human liver microsomes (n=17) in relation to CYP-dependent ATRA 4-hydroxylation. Observed rates of ATRA cooxidation (4.6 - 20 pmol mg protein(-1) min(-1)) and 4 hydroxylation (8.7 - 45 pmol mg protein(-1) min(-1)) were quantitatively similar and exhibited similar individual variation (4 and 5 fold, respectively). From kinetic studies cooxidation was an efficient process in human hepatic microsomes (V(max) K(m)(-1)=0.25) compared with NADPH- and NADH-mediated 4-hydroxylation by CYP (V(max) K(m)(-1)=0.14 and 0.02, respectively). The capacity of lipid hydroperoxide metabolites of arachidonic acid to mediate ATRA oxidation was established directly, but downstream products (D, E, F and I-series prostaglandins) were inactive. cDNA-expressed CYPs supported ATRA oxidation by lipid hydroperoxides. Whereas CYPs 2C8, 2C9 and 3A4, but not CYPs 1A2 or 2E1, were effective catalysts of the NADPH-mediated reaction, cooxidation supported by 15(S)-hydroperoxyeicosatetraenoic acid was mediated by all five CYPs. The cooxidation reaction in human hepatic microsomes was inhibited by the CYP inhibitor miconazole. These findings indicate that ATRA oxidation is quantitatively significant in human liver. Lipid hydroperoxides generated by intracellular enzymes such as prostaglandin synthase and lipoxygenases are sources of activated oxygen for CYP-mediated deactivation of ATRA to polar products. PMID- 11030739 TI - Corrigendum PMID- 11030738 TI - The interaction between components of the fibrinolytic system and GPIb/V/IX of platelets thrombus formation in mice. AB - The interaction of fibrinolytic components with GPIb/V/IX of platelets on thrombus formation, was investigated in mice deficient in tissue type (tPA-/-), urokinase type plasminogen activator (uPA-/-) or plasminogen activator inhibitor 1 (PAI-1-/-) and in their wild type control (tPA+/+, uPA+/+, PAI-1+/+). A thrombus was induced in the murine carotid artery using a photochemical reaction. The times to occlusion after the initiation of endothelial injury in all wild type mice was within 12 min, and no significant changes in occlusion delay were observed in uPA-/- and tPA-/- mice compared to wild type mice, whereas that of PAI-1 mice were significantly prolonged (16.9+/-2.9 min, P<0.05). When high molecular weight aurintricarboxylic acid (ATA), an inhibitor of platelet glycoprotein Ib/V/IX, was administered, the time to occlusion was prolonged in a dose-dependent manner in all types of mice. However, when this compound was injected in tPA-/- mice, the most significant changes were observed: i.e. the estimated ED(50) was 20.2 times higher than that in tPA+/+ mice, but the estimated ED(50) in uPA-/- mice was not changed as compared with that of wild type mice. On the other hand, when ATA was injected in PAI-1-/- mice, the estimated ED(50) was significantly decreased (P<0.05). Platelet aggregation induced by botrocetin was not significantly different among all types of mice. The bleeding time was prolonged in a dose dependent-manner when ATA was injected in all types of mice. In conclusion, the antithrombotic effect of inhibition of platelet GPIb/V/IX is severely affected by the absence or presence of tPA production on thrombus formation and the inhibition of PAI-1 could enhance this antithrombotic effect. PMID- 11030740 TI - Corrigendum PMID- 11030741 TI - Keratan sulfate: structure, biosynthesis, and function. AB - The last 5 years have seen a marked increase in research on keratan sulfate (KS) and a concomitant increase in our understanding of the range of molecules that carry this adaptable polysaccharide. More than 15 KS-linked proteins have been identified and many of the genes encoding these have been cloned. KS-containing molecules have been identified in numerous epithelial and neural tissues in which KS expression responds to embryonic development, physiological variations, and to wound healing. A corneal cell culture system has been developed in which long term KS biosynthesis is maintained. Progress has been made toward identification of the glycosyl- and sulfotransferases responsible for KS biosynthesis. A mouse knockout of a corneal KS-proteoglycan has provided the first experimental support for the role of KS in corneal transparency. Evidence has also been presented supporting functional roles of KS in cellular recognition of protein ligands, axonal guidance, cell motility, and in embryo implantation. These findings have served to expand the concept of what keratan sulfate is and the potential roles it may play in the cellular biology of diverse tissues. PMID- 11030742 TI - Molecular dynamics simulation of human prion protein including both N-linked oligosaccharides and the GPI anchor. AB - Although glycosylation appears to protect prion protein (PrP(C)) from the conformational transition to the disease-associated scrapie form (PrP(Sc)), available NMR structures are for non-glycosylated PrP(C), only. To investigate the influence of both the two N-linked glycans, Asn181 and Asn197, and of the GPI anchor attached to Ser230, on the structural, dynamical and electrostatic behavior of PrP, we have undertaken molecular dynamics simulations on the C terminal region of human prion protein HU:PrP(90-230), with and without the three glycans. The simulations used the AMBER94 force field in a periodic box model with explicit water molecules, considering all long-range electrostatic interactions. The results suggest the structured part of the protein, HU:PrP(127 227) is stabilized overall from addition of the glycans, specifically by extensions of Helix-B and Helix-C and reduced flexibility of the linking turn containing Asn197, although some regions such as residues in the turn (165-170) between Strand-B and Helix-B have increased flexibility. The stabilization appears indirect, by reducing the mobility of the surrounding water molecules, and not from specific interactions such as H bonds or ion pairs. The results are consistent with glycosylation at Asn197 having a stabilizing role, while that at Asn181, in a region with already stable secondary structure, having a more functional role, in agreement with literature suggestions. Due to three negatively charged SiaLe(x) groups per N-glycan, the surface electrostatic properties change to a negative electrostatic field covering most of the C terminal part, including the surface of Helix-B and Helix-C, while the positively charged N-terminal part PrP(90-126) of undefined structure creates a positive potential. The unusual hydrophilic Helix-A (144-152) is not covered by either of these dominant electrostatic fields, and modeling shows it could readily dimerize in anti parallel fashion. In combination with separate simulations of the GPI anchor in a membrane model, the results show the GPI anchor is highly flexible and would maintain the protein at a distance between 9 and 13 A from the membrane surface, with little influence on its structure or orientational freedom. PMID- 11030743 TI - A strain of human influenza A virus binds to extended but not short gangliosides as assayed by thin-layer chromatography overlay. AB - A human strain of influenza virus (A, H1N1) was shown to bind in an unexpected way to leukocyte and other gangliosides when compared with avian virus (A, H4N6) as assayed on TLC plates. The human strain bound only to species with about 10 or more sugars, while the avian strain bound to a wide range of gangliosides including the 5-sugar gangliosides. By use of specific lectins, antibodies, and FAB and MALDI-TOF mass spectrometry an attempt was done to preliminary identify the sequences of leukocyte gangliosides recognized by the human strain. The virus binding pattern did not follow binding by VIM-2 monoclonal antibody and was not identical with binding by anti-sialyl Lewis x antibody. There was no binding by the virus of linear NeuAcalpha3- or NeuAcalpha6-containing gangliosides with up to seven monosaccharides per mol of ceramide. Active species were minor NeuAcalpha6-containing molecules with probably repeated HexHexNAc units and fucose branches. This investigation demonstrates marked distinctions in the recognition of gangliosides between avian and human influenza viruses. Our data emphasize the importance of structural factors associated with more distant parts of the binding epitope and the complexity of carbohydrate recognition by human influenza viruses. PMID- 11030744 TI - Dolichol phosphate mannose synthase from the filamentous fungus Trichoderma reesei belongs to the human and Schizosaccharomyces pombe class of the enzyme. AB - Dolichol phosphate mannose (DPM) synthase activity, which is required in N: glycosylation, O-mannosylation, and glycosylphosphatidylinositol membrane anchoring of protein, has been postulated to regulate the Trichoderma reesei secretory pathway. We have cloned a T.reesei cDNA that encodes a 243 amino acid protein whose amino acid sequence shows 67% and 65% identity, respectively, to the Schizosaccharomyces pombe and human DPM synthases, and which lacks the COOH terminal hydrophobic domain characteristic of the Saccharomyces cerevisiae class of synthase. The Trichoderma dpm1 (Trdpm1) gene complements a lethal null mutation in the S.pombe dpm1(+) gene, but neither restores viability of a S.cerevisiae dpm1-disruptant nor complements the temperature-sensitivity of the S. cerevisiae dpm1-6 mutant. The T.reesei DPM synthase is therefore a member of the "human" class of enzyme. Overexpression of Trdpm1 in a dpm1(+)::his7/dpm1(+) S.pombe diploid resulted in a 4-fold increase in specific DPM synthase activity. However, neither the wild type T. reesei DPM synthase, nor a chimera consisting of this protein and the hydrophobic COOH terminus of the S.cerevisiae DPM synthase, complemented an S.cerevisiae dpm1 null mutant or gave active enzyme when expressed in E.coli. The level of the Trdpm1 mRNA in T.reesei QM9414 strain was dependent on the composition of the culture medium. Expression levels of Trdpm1 were directly correlated with the protein secretory capacity of the fungus. PMID- 11030745 TI - Stereochemical metrics of lectin-carbohydrate interactions: comparison with protein-protein interfaces. AB - A global census of stereochemical metrics including interface size, hydropathy, amino acid propensities, packing and hydrogen bonding was carried out on 32 x-ray elucidated structures of lectin-carbohydrate complexes covering eight different lectin families. It is shown that the interactions at primary binding subsites are more efficient than at other subsites. Another salient behavior found for primary subsites was a marked negative correlation between the interface size and the polar surface content. It is noteworthy that this demographic rule is delineated by lectins with unrelated phylogenetic origin, indicating that independent interface architectures have evolved through common optimization paths. The structural properties of lectin-carbohydrate interfaces were compared with those characterizing a set of 32 protein homodimers. Overall, the analysis shows that the stereochemical bases of lectin-carbohydrate and protein-protein interfaces differ drastically from each other. In comparison with protein-protein complexes, lectin-carbohydrate interfaces have superior packing efficiency, better hydrogen bonding stereochemistry, and higher interaction cooperativity. A similar conclusion holds in the comparison with protein-protein heterocomplexes. We propose that the energetic consequence of this better interaction geometry is a larger decrease in free energy per unit of area buried, feature that enables lectins and carbohydrates to form stable complexes with relatively small interface areas. These observations lend support to the emerging notion that systems differing from each other in their stereochemical metrics may rely on different energetic bases. PMID- 11030746 TI - A novel variant form of murine beta-1, 6-N-acetylglucosaminyltransferase forming branches in poly-N-acetyllactosamines. AB - A novel form of murine beta-1,6-N:-acetylglucosaminyltransferase that forms branches in poly-N:-acetyllactosamines (designated as IGnT B) was cloned based on sequence homology to the known IGnT (designated as IGnT A). When expressed as proteins, IGnT B showed higher specific activity than IGnT A. The C-terminal 1/4 of IGnT B was identical to that of IGnT A, while the rest of the predicted sequences showed 63% identity. Genomic analysis indicated that IGnT A and IGnT B were derived by alternative splicing; the unique portion was encoded by exon 1, and the common portion was encoded by exons 2 and 3. IGnT B showed an expression profile closely related to that of IGnT A and was strongly expressed in the liver, kidney and intestine, and moderately in the mammary gland, submaxially gland, embryonic stem cells, and embryonal carcinoma cells. The specificity of IGnT B examined using various substrates was indistinguishable from that of IGnT A, which is classified as the central acting IGnT (cIGnT). Thus, IGnT B acted on Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc, but not on GlcNAcbeta1-3Galbeta1-4Glc. It formed branches in both of the internal galactosyl residues of Galbeta1-4Glc NAcbeta1-3Galbeta1-4GlcNAcbeta1-++ +3Galbeta1-4Glc, and prolonged incubation resulted in production of the di-branched oligosaccharide. Although addition of sialic acid to the terminal galactosyl residue did not abolish the acceptor activity, alpha2-6 sialylation was a preferred one as compared to alpha2-3 sialylation. PMID- 11030747 TI - The type and yield of lipopolysaccharide from symbiotically deficient rhizobium lipopolysaccharide mutants vary depending on the extraction method. AB - At least 18 lipopolysaccharide (LPS) extraction methods are available, and no single method is universally applicable. Here, the LPSs from four R.etli, one R.leguminosarum bv. trifolii mutant, 24AR, and the R.etli parent strain, CE3, were isolated by hot phenol/water (phi;/W), and phenol/EDTA/triethylamine (phi/EDTA/TEA) extraction. The LPS in various preparations was quantified, analyzed by deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE), and by immunoblotting. These rhizobia normally have two prominent LPS forms: LPS I, which has O-polysaccharide, and LPS II, which has none. The LPS forms obtained depend on the method of extraction and vary depending on the mutant that is extracted. Both methods extract LPS I and LPS II from CE3. The phi/EDTA/TEA, but not the phi/W, method extracts LPS I from mutants CE358 and CE359. Conversely, the phi;/W but not the phi;/EDTA/TEA method extracts CE359 LPS V, an LPS form with a truncated O-polysaccharide. phi/EDTA/TEA extraction of mutant CE406 gives good yields of LPS I and II, while phi/W extraction gives very small amounts of LPS I. The LPS yield from all the strains using phi/EDTA/TEA extraction is fairly consistent (3-fold range), while the yields from phi/W extraction are highly variable (850-fold range). The phi/EDTA/TEA method extracts LPS I and LPS II from mutant 24AR, but the phi/W method partitions LPS II exclusively into the phenol phase, making its recovery difficult. Overall, phi/EDTA/TEA extraction yields more forms of LPS from the mutants and provides a simpler, faster, and less hazardous alternative to phi/W extraction. Nevertheless, it is concluded that careful analysis of any LPS mutant requires the use of more than one extraction method. PMID- 11030749 TI - Preparation and structural determination of dermatan sulfate-derived oligosaccharides. AB - Eight oligosaccharides were prepared from dermatan sulfate (DS) and their structures were elucidated. Porcine intestinal mucosal DS was subjected to controlled depolymerization using chondroitin ABC lyase (chondroitinase ABC). The oligosaccharide mixture formed was fractionated by low-pressure gel permeation chromatography (GPC). Size uniform mixtures of disaccharides, tetrasaccharides, hexasaccharides, octasaccharides, decasaccharides, and dodecasaccharides were obtained. Each size-fractionated mixture was then purified on the basis of charge by repetitive semi-preparative strong-anion-exchange (SAX) high-performance liquid chromatography (HPLC). This approach has led to the isolation of six homogeneous oligosaccharides. The size of the oligosaccharides were determined using GPC-HPLC. Treatment of tetrasaccharide and hexasaccharide fragments with Hg(OAc)2 afforded trisaccharide and pentasaccharide products, respectively. The purity of the oligosaccharides obtained was confirmed by analytical SAX-HPLC, and capillary electrophoresis (CE). The molecular mass and degree of sulfation of the eight purified oligosaccharides were elucidated using electrospray ionization (ESI) mass spectrometry and their structures were established with high field nuclear magnetic resonance (NMR) spectroscopy. These DS-oligosaccharides are currently being used to study for interaction of the DS with biologically important proteins. PMID- 11030748 TI - Evidence supporting a late Golgi location for lactosylceramide to ganglioside GM3 conversion. AB - Ganglioside GM2 synthase and other enzymes required for complex ganglioside synthesis were localized recently to the trans Golgi network (TGN). However, there are conflicting reports as to the location of GM3 synthase; originally this enzyme was detected in the early Golgi of rat liver but a recent report localized it to the late Golgi. We have used chimeric forms of ganglioside GM2 synthase to determine if the location of lactosylceramide (LacCer) to GM3 conversion in Chinese hamster ovary (CHO) cells was the early or late Golgi. Our approach tested whether GM3 could be utilized as a substrate by GM2 synthase chimeras which were targeted to compartments earlier than the trans Golgi, i.e., GM3 produced in the cis Golgi should be utilized by GM2 synthase located anywhere in the Golgi whereas GM3 produced in the trans Golgi should only be used by GM2 synthase located in the trans Golgi or TGN. Comparison of cell lines stably expressing these chimeras revealed that the in vivo functional activity of GM2 synthase decreased progressively as the enzyme was targeted to earlier compartments; specifically, the percentage of GM3 converted to GM2 was 83-86% for wild type enzyme, 70% for the medial Golgi targeted enzyme, 13% for the ER and cis Golgi targeted enzyme, and only 1.7% for the ER targeted enzyme. Thus, these data are consistent with a late Golgi location for LacCer to GM3 conversion in these cells. PMID- 11030750 TI - Functional expression of Escherichia coli enzymes synthesizing GDP-L-fucose from inherent GDP-D-mannose in Saccharomyces cerevisiae. AB - Fucosylation of glycans on glycoproteins and -lipids requires the enzymatic activity of relevant fucosyltransferases and GDP-L-fucose as the donor. Due to the biological importance of fucosylated glycans, a readily accessible source of GDP-L-fucose would be required. Here we describe the construction of a stable recombinant S.cerevisiae strain expressing the E.coli genes gmd and wcaG encoding the two enzymes, GDP-mannose-4,6-dehydratase (GMD) and GDP-4-keto-6-deoxy-D mannose-3,5-epimerase/4-reductase (GMER(FX)) respectively, needed to convert GDP mannose to GDP-fucose via the de novo pathway. Taking advantage of the rich inherent cytosolic GDP-mannose pool in S.cerevisiae cells we could easily produce 0.2 mg/l of GDP-L-fucose with this recombinant yeast strain without addition of any external GDP-mannose. The GDP-L-fucose product could be used as the fucose donor for alpha1,3fucosyltransferase to synthesize sialyl Lewis x (sLex), a glycan crucial for the selectin-dependent leukocyte traffic. PMID- 11030751 TI - Biosynthetic incorporation of unnatural sialic acids into polysialic acid on neural cells. AB - In this study we demonstrate that polysialyltransferases are capable of accepting unnatural substrates in terminally differentiated human neurons. Polysialyltransferases catalyze the glycosylation of the neural cell adhesion molecule (NCAM) with polysialic acid (PSA). The unnatural sialic acid analog, N levulinoyl sialic acid (SiaLev), was incorporated into cell surface glycoconjugates including PSA by the incubation of cultured neurons with the metabolic precursor N-levulinoylmannosamine (ManLev). The ketone group within the levulinoyl side chain of SiaLev was then used as a chemical handle for detection using a biotin probe. The incorporation of SiaLev residues into PSA was demonstrated by protection from sialidases that can cleave natural sialic acids but not those bearing unnatural N-acyl groups. The presence of SiaLev groups on the neuronal cell surface did not impede neurite outgrowth or significantly affect the distribution of PSA on neuronal compartments. Since PSA is important in neural plasticity and development, this mechanism for modulating PSA structure might be useful for functional studies. PMID- 11030752 TI - Mucolipidosis type IV is caused by mutations in a gene encoding a novel transient receptor potential channel. AB - Mucolipidosis type IV (MLIV) is a developmental neurodegenerative disorder characterized by severe neurologic and ophthalmologic abnormalities. The MLIV gene, ML4 (MCOLN1), has recently been localized to chromosome 19p13.2-13.3 by genetic linkage. Here we report the cloning of a novel transient receptor potential cation channel gene and show that this gene is mutated in patients with the disorder. ML4 encodes a protein, which we propose to call mucolipin, which has six predicted transmembrane domains and is a member of the polycystin II subfamily of the Drosophila transient receptor potential gene family. The role of a potential receptor-stimulated cation channel defect in the pathogenesis of mucolipidosis IV is discussed. PMID- 11030753 TI - The neuronal apoptosis inhibitory protein suppresses neuronal differentiation and apoptosis in PC12 cells. AB - The human neuronal apoptosis inhibitory protein (NAIP) gene has been discovered as a candidate gene for spinal muscular atrophy, a genetic disorder characterized by motor neuron loss in the spinal cord. The telomeric NAIP gene on human chromosome 5 is deleted together with survival motor neurons (SMN) in many cases of the most severe forms of the disorder. NAIP, c-IAP1 (inhibitor of apoptosis 1), c-IAP2, X-IAP, survivin and Apollon comprise the mammalian inhibitors of the apoptosis family and contain an N-terminal domain with 1-3 imperfect repeats of an approximately 65 amino acids domain named the baculovirus IAP repeat (BIR) motif. We identified six NAIP genes in the mouse genome which were found to be expressed in a broad range of tissues. Furthermore, we have investigated the effects of NAIP in the rat pheochromocytoma PC12 cell line. These cells differentiate in the presence of nerve growth factor (NGF) into cells that resemble sympathetic neurons. We observed that NAIP overexpression impaired NGF induced neurite outgrowth. The BIR motifs of NAIP (residues 1-345) were not required for this effect. However, the BIR domains of NAIP were essential to prevent apoptosis in PC12 cells after NGF deprivation or TNF-alpha receptor stimulation. Expression of full-length but not BIR-deleted-NAIP protects against cell death. This correlates with reduced activity of the cell death effector protease, caspase-3, in lysates of NAIP-PC12 cells, as measured by cleavage of the fluorogenic tetrapeptide substrate Asp-Glu-Val-Asp. Thus, unregulation of cellular differentiation and/or caspase suppression may contribute to motoneuron dysfunction and cell death in spinal muscular atrophy where NAIP is mutated. PMID- 11030754 TI - Expanded polyglutamines induce neurodegeneration and trans-neuronal alterations in cerebellum and retina of SCA7 transgenic mice. AB - Among the eight progressive neurodegenerative diseases caused by polyglutamine expansions, spinocerebellar ataxia type 7 (SCA7) is the only one to display degeneration in both brain and retina. We show here that mice overexpressing full length mutant ataxin-7[Q90] either in Purkinje cells or in rod photoreceptors have deficiencies in motor coordination and vision, respectively. In both models, although with different time courses, an N-terminal fragment of mutant ataxin-7 accumulates into ubiquitinated nuclear inclusions that recruit a distinct set of chaperone/proteasome subunits. A severe degeneration is caused by overexpression of ataxin-7[Q90] in rods, whereas a similar overexpression of normal ataxin 7[Q10] has no obvious effect. The degenerative process is not limited to photoreceptors, showing secondary alterations of post-synaptic neurons. These findings suggest that proteolytic cleavage of mutant ataxin-7 and trans-neuronal responses are implicated in the pathogenesis of SCA7. PMID- 11030755 TI - Mdx mice inducibly expressing dystrophin provide insights into the potential of gene therapy for duchenne muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is an X-linked recessive disease caused by the lack of expression of the dystrophin protein in muscle tissues. We genetically engineered a mouse model (mdx) of DMD that allowed for the high level and inducible transcription of a dystrophin mini-gene. This was achieved via the tetracycline-responsive transactivator (tTA) system. Multiple analyses confirmed that dystrophin expression in the mice was: (i) tTA dependent; (ii) correctly localized to the sarcolemmal membranes; (iii) capable of preventing the onset of dystrophy; and (iv) effectively blocked by the oral administration of tetracyclines. The model allowed us to somatically extinguish or induce dystrophin gene transcription. Somatic induction of dystrophin transcription prevented the onset of muscular dystrophy in some muscle groups. The levels of phenotypic rescue were influenced, however, by the age of the animals at the time of dystrophin induction. We also found that despite somatic termination of dystrophin gene transcription, the dystrophin protein was found to be associated with the sarcolemmal membrane for at least 26 weeks. Persistent detection of dystrophin was also accompanied by a prolonged protection of the muscle cells from the onset of dystrophy. The findings demonstrated that somatic transfer of the dystrophin gene not only may allow for the prevention of muscular dystrophy in multiple muscle groups, but also may be accompanied by persistent efficacy, secondary to the long-term functional stability of the dystrophin protein in vivo. This model should be useful in future studies concerning the potential of genetic therapy for DMD, as well as other muscle disorders. PMID- 11030756 TI - Interaction between the G1057D variant of IRS-2 and overweight in the pathogenesis of type 2 diabetes. AB - The insulin receptor substrate-2 (IRS-2) is a major insulin signalling molecule. IRS-2 inactivation in mice induces a form of diabetes characterized by peripheral insulin resistance and reduced beta cell mass. We tested the hypothesis that a common non-conservative amino acid substitution of IRS-2 (G1057D) might interact with overweight in the pathogenesis of type 2 diabetes. The variant was genotyped in 193 Italian patients with type 2 diabetes and 206 control subjects. In the absence of overweight, the risk of type 2 diabetes decreased according to the dosage of the D1057 allele (odds ratio for GD genotype 0.46 [95% CI 0.25-0.86]; DD genotype 0.18 [0.04-0.68]; P for trend = 0.0012). Conversely, the interaction between overweight and genotype increased the risk of type 2 diabetes according to the dosage of the D1057 allele (odds ratio for GD genotype 2.50 [1.11-5.65]; DD genotype 5.74 [1.11-29. 78]; P for trend = 0.0047). Among controls, fasting C peptide levels, after adjustment for plasma glucose, were inversely related to the dosage of the D1057 allele (P = 0.020). This finding suggested that carriers of the D1057 allele may have higher insulin sensitivity and supported the protective effect of this allele. Conversely, among overweight patients there was a parallel increase in fasting plasma glucose (P for trend = 0.037) and fasting C peptide according to the dosage of the D1057 allele, suggesting that higher insulin resistance and relative beta cell failure contributed to the increased risk of type 2 diabetes in overweight carriers of this allele. These data provide evidence for a strong association between type 2 diabetes and the G1057D common genetic variant of IRS-2, which appears to be protective against type 2 diabetes in a codominant fashion. Overweight appears to modify the effect of this polymorphism toward a higher risk of type 2 diabetes. Carriers of this polymorphism may represent an elective target for prevention of type 2 diabetes through preventing or treating excessive weight. PMID- 11030757 TI - Human frataxin maintains mitochondrial iron homeostasis in Saccharomyces cerevisiae. AB - Frataxin is a nuclear-encoded mitochondrial protein widely conserved among eukaryotes. Human frataxin (fxn) is severely reduced in Friedreich ataxia (FRDA), a frequent autosomal recessive neuro- and cardio-degenerative disease. Whereas the function of fxn is unknown, the yeast frataxin homolog (Yfh1p) has been shown to be involved in mitochondrial iron homeostasis and protection from free radical toxicity. Evidence of iron accumulation and oxidative damage in cardiac tissue from FRDA patients suggests that fxn may have a similar function, but whether yeast and human frataxin actually have interchangeable roles in mitochondrial iron homeostasis is unknown. We show that a wild-type FRDA cDNA can complement Yfh1p-deficient yeast (yfh1 delta) by preventing the mitochondrial iron accumulation and oxidative damage associated with loss of Yfh1p. We analyze the functional effects of two FRDA point mutations, G130V and W173G, associated with a mild and a severe clinical presentation, respectively. The G130V mutation affects protein stability and results in low levels of mature (m) fxn, which are nevertheless sufficient to rescue yfh1 delta yeast. The W173G mutation affects protein processing and stability and results in severe m-fxn deficiency. Expression of the FRDA (W173G) cDNA in yfh1 delta yeast leads to increased levels of mitochondrial iron which are not as elevated as in Yfh1p-deficient cells but are above the threshold for oxidative damage of mitochondrial DNA and iron-sulfur centers, causing a typical yfh1 delta phenotype. These results demonstrate that fxn functions like Yfh1p, providing experimental support to the hypothesis that FRDA is a disorder of mitochondrial iron homeostasis. PMID- 11030758 TI - Pleiotropic effects of the melanocortin 1 receptor (MC1R) gene on human pigmentation. AB - Variants of the melanocortin 1 receptor (MC1R) gene are common in individuals with red hair and fair skin, but the relative contribution to these pigmentary traits in heterozygotes, homozygotes and compound heterozygotes for variants at this locus from the multiple alleles present in Caucasian populations is unclear. We have investigated 174 individuals from 11 large kindreds with a preponderance of red hair and an additional 99 unrelated redheads, for MC1R variants and have confirmed that red hair is usually inherited as a recessive characteristic with the R151C, R160W, D294H, R142H, 86insA and 537insC alleles at this locus. The V60L variant, which is common in the population may act as a partially penetrant recessive allele. These individuals plus 167 randomly ascertained Caucasians demonstrate that heterozygotes for two alleles, R151C and 537insC, have a significantly elevated risk of red hair. The shade of red hair frequently differs in heterozygotes from that in homozygotes/compound heterozygotes and there is also evidence for a heterozygote effect on beard hair colour, skin type and freckling. The data provide evidence for a dosage effect of MC1R variants on hair as well as skin colour. PMID- 11030759 TI - Dramatic mutation instability in HD mouse striatum: does polyglutamine load contribute to cell-specific vulnerability in Huntington's disease? AB - An unstable CAG triplet repeat expansion encoding a polyglutamine stretch within the ubiquitously expressed protein huntingtin is responsible for causing Huntington's disease (HD). By quantifying the repeat sizes of individual mutant alleles in tissues derived from an accurate genetic mouse model of HD we show that the mutation becomes very unstable in striatal tissue. The expansion-biased changes increase with age, such that some striatal cells from old HD mice contain mutations that have tripled in size. If this pattern of repeat instability is recapitulated in human striatal tissue, the concomitant increased polyglutamine load may contribute to the patterns of selective neuronal cell death in HD. Our findings also suggest that trinucleotide repeat instability can occur by mechanisms that are not replication-based. PMID- 11030760 TI - Intramuscular injection of a plasmid vector expressing human apolipoprotein E limits progression of xanthoma and aortic atheroma in apoE-deficient mice. AB - Apolipoprotein-E (apoE) protects against coronary artery disease via hepatic removal of atherogenic remnant lipoproteins, sequestration of cholesterol from vessel walls and local anti-oxidant, anti-platelet and anti-inflammatory actions. ApoE gene transfer may thus ameliorate a hyperlipidaemic profile and have beneficial effects at lesion sites to prevent or regress atherosclerosis, a concept endorsed by adenoviral-mediated hepatic expression studies. Here, using plasmid vectors expressing allelic human apoE2 or apoE3 isoforms, skeletal muscle was evaluated as an effective secretory platform for apoE gene augmentation. Transfected myoblasts and myotubes were found to efficiently secrete recombinant apoE in vitro as spherical 10-16 nm lipoprotein particles with pre-beta mobility. Intramuscular plasmid injection in apoE(-/-) mice, which develop spontaneous atherosclerotic plaque and xanthoma resulted in expression and secretion of apoE. Human apoE mRNA was detected by RT-PCR in injected muscles and, although concentrations of apoE3, which is rapidly cleared from plasma, were near ELISA detection limits, levels of plasma apoE2 were measurable (17.5 +/- 4.3 ng/ml). To assess whether muscle-based expression of apoE2 could inhibit atherogenesis, long term follow-up studies were conducted. Although hyperlipidaemia was not reduced in treated animals, end-point pathology showed clear retardation of atherosclerotic and xanthomatous lesions. Up to 9 months following a single apoE2 plasmid administration, atherosclerotic lesion coverage in proximal aorta was significantly reduced by 20-30% (P < 0.01), whereas development of gross dorsal xanthoma (>5 mm diameter) was effectively reduced to zero. We conclude that expression of apoE from ectopic muscle sites has therapeutic potential to limit progression of atherosclerosis. PMID- 11030761 TI - New mutations in MID1 provide support for loss of function as the cause of X linked Opitz syndrome. AB - Opitz syndrome (OS) is a genetically heterogeneous malformation disorder. Patients with OS may present with a variable array of malformations that are indicative of a disturbance of the primary midline developmental field. Mutations in the C-terminal half of MID1, an RBCC (RING, B-box and coiled-coil) protein, have recently been shown to underlie the X-linked form of OS. Here we show that the MID1 gene spans at least 400 kb, almost twice the distance originally reported and has a minimum of six mRNA isoforms as a result of the alternative use of 5' untranslated exons. In addition, our detailed mutational analysis of MID1 in a cohort of 15 patients with OS has resulted in the identification of seven novel mutations, two of which disrupt the N-terminus of the protein. The most severe of these (E115X) is predicted to truncate the protein before the B box motifs. In a separate patient, a missense change (L626P) was found that also represents the most C-terminal alteration reported to date. As noted with other C terminal mutations, GFP fusion constructs demonstrated that the L626P mutant formed cytoplasmic clumps in contrast to the microtubular distribution seen with the wild-type sequence. Notably, however, both N-terminal mutants showed no evidence of cytoplasmic aggregation, inferring that this feature is not pathognomonic for X-linked OS. These new data and the finding of linkage to MID1 in the absence of a demonstrable open reading frame mutation in a further family support the conclusion that X-linked OS results from loss of function of MID1. PMID- 11030762 TI - Two long homologous retroviral sequence blocks in proximal Yq11 cause AZFa microdeletions as a result of intrachromosomal recombination events. AB - We mapped the breakpoints of the AZoospermia factor a (AZFa) microdeletion located in proximal Yq11 in six men with complete germ cell aplasia, i.e. Sertoli Cell Only syndrome (SCO). The proximal breakpoints were identified in a long retroviral sequence block (HERV15yq1: 9747 nucleotides) at the 5' end of the DYS11 DNA locus in Yq11, interval D3. The distal breakpoints were found in a homologous HERV15 sequence block mapped to the Yq11 interval D6, i.e. in the distal part of the AZFa region (HERV15yq2: 9969 nucleotides). Compared with the HERV15yq1 sequence, HERV15yq2 is marked by a deletion of a HERV15 sequence domain at its 5' end and insertion of an LINE 1 3'-UTR sequence block (L1PA4) of similar length at its 3' end. The deletion of the L1PA4 element was recognized as the molecular origin of the DYS11 12f2 restriction fragment length polymorphism. For all six AZFa patients it was possible to perform PCR experiments bridging both retroviral sequence blocks, which map in a distance of 781.557 kb in proximal Yq11 in fertile men. The AZFa breakpoint-fusion regions were located in their recombined HERV15yq1/HERV15yq2 sequence blocks in either one of two long identical sequence domains (ID1 and ID2). We therefore assume that intrachromosomal recombination events between the two homologous retroviral sequence blocks in proximal Yq11 are probably the causative agents for most of the AZFa microdeletions observed in men with SCO syndrome. A mean value of 792 kb was estimated for their molecular lengths. PMID- 11030764 TI - News and notes: october 2000 PMID- 11030763 TI - Topical sucralfate for erosive irritant diaper dermatitis. PMID- 11030765 TI - Bullous pemphigoid treated with leflunomide: a novel immunomodulatory agent. PMID- 11030766 TI - Occult neurofibroma and increased S100 protein in the skin of patients with neurofibromatosis type 1: new insight to the etiopathomechanism of neurofibromas. AB - BACKGROUND: Neurofibromas represent proliferation of the connective tissue cells of peripheral nerves and deposition of collagenous extracellular matrix. There is evidence that the appearance and growth of neurofibromas may be associated with prior or ongoing mechanical trauma in patients with neurofibromatosis type 1 (NF1). OBJECTIVE: To study the histologic characteristics of apparently healthy skin of patients with NF1. DESIGN: The histologic features of healthy-looking skin of patients with NF1 were analyzed. SETTING: University hospital. PATIENTS: Ten patients who fulfilled the criteria for NF1. INTERVENTIONS: Punch biopsy specimens of healthy-looking skin of the forearm from 9 volunteer patients and of the upper eyelid during cosmetic operation from 1 volunteer patient were obtained. MAIN OUTCOME MEASURES: The main outcomes were not predicted, and the hypothesis was formulated during data collection. RESULTS: Apparently unaffected skin of 5 patients with NF1 was studied by routine histologic testing with respect to expression of S100 protein. Unexpectedly, analysis of the samples revealed the presence of a small neurofibroma tumor in one of the samples. The tumor was located in deep dermis around a hair follicle. In addition, neurofibromatous tissue not large enough to be called a tumor was found on the same anatomical location in another patient. In further studies, 10 punch biopsy specimens of apparently healthy skin from patients with NF1 were similarly sectioned and analyzed. No tumors were found in these additional samples. In 4 patients, however, abundant S100 protein-positive cells were located within collagenous extracellular matrix surrounding hair follicles. CONCLUSIONS: The skin of patients with NF1 might be more widely affected than previously thought and occult neurofibromas are not rare. PMID- 11030767 TI - The natural history of chronic actinic dermatitis. AB - OBJECTIVE: To determine the prognosis for resolution of abnormal cutaneous photosensitivity in patients with chronic actinic dermatitis (also known as the photosensitivity dermatitis and actinic reticuloid syndrome). DESIGN: Historical cohort study involving follow-up of patients for up to 24 years from diagnosis. SETTING: A Scottish tertiary referral center for investigation of photodermatosis. PATIENTS: One hundred seventy-eight patients with chronic actinic dermatitis, 62% of a cohort of 285 living patients identified in the Photobiology Unit database. INTERVENTIONS: Recall for repeated clinical assessment and monochromator phototesting. All patients underwent patch testing when initially assessed; this was repeated at follow-up in a subgroup of patients. MAIN OUTCOME MEASURES: Resolution of abnormal photosensitivity, defined as clinical resolution and return of phototest responses to within normal population limits. In addition, possible prognostic factors for resolution of photosensitivity were examined. RESULTS: The probability of abnormal photosensitivity resolving by 10 years from diagnosis is 1 in 5. Particularly severe abnormal UV-B photosensitivity (minimal erythema dose at 305+/-5 nm half maximum bandwidth, < or =5.6 mJ x cm(-2)) and the identification of separate contact allergens in 2 or more patch test batteries are predictors of a poorer prognosis for resolution. Loss of contact allergies was not associated with a different prognosis for photosensitivity resolution. Our findings probably underestimate the probability of resolution, as those referred to a tertiary referral center and willing to attend for follow-up may include a disproportionate number of severely affected patients. CONCLUSIONS: Newly diagnosed patients can be told that most of them will improve with appropriate UV/visible light and allergen avoidance and that there is hope that their photosensitivity will completely resolve. PMID- 11030768 TI - Tissue-engineered skin (Apligraf) in the healing of patients with epidermolysis bullosa wounds. AB - BACKGROUND: At present, wound treatment of inherited epidermolysis bullosa (EB) is only supportive. OBJECTIVE: To determine the safety and clinical effects of tissue-engineered skin (Apligraf; Organogenesis Inc, Canton, Mass) in the healing of wounds of patients with different types of EB. DESIGN: An open-label uncontrolled study of 15 patients with EB treated with tissue-engineered skin. Each patient received tissue-engineered skin on up to 2 wounds on each of 3 clinic visits: day 1, week 6, and week 12. They were evaluated 7 (+/- 3) days and 6 weeks after each round of treatment. A quality-of-life survey was administered during week 6. SETTING: University of Miami, Miami, Fla. PATIENTS: Volunteers with EB. MAIN OUTCOME MEASURE: Safety and wound healing. RESULTS: A total of 69 different acute wounds received tissue-engineered skin at the day-1 (24 wounds), week-6 (23 wounds), and week-12 (22 wounds) visits. Overall, 63 wounds (79%) were found healed at the day-7 visit. Of the acute wounds, 82% (51/62) were healed 6 weeks after being treated, 75% (27/36) after 12 weeks, and 79% (11/14) after 18 weeks. Nine chronic wounds were also treated. Four were healed at 6 weeks; however, 7 were still open at the last clinic visit (week 18). There were no signs of rejection or clinical infection and no adverse events related to the tissue-engineered skin. The quality of life for most patients improved after treatment. Compared with patients' recollection of wounds treated with standard dressings, healing was faster and less painful. CONCLUSION: In this series of patients, tissue-engineered skin induced very rapid healing, was not clinically rejected, and was devoid of adverse effects. It was felt by the patients and families to be more effective than conventional dressings for EB wounds. PMID- 11030769 TI - Isotretinoin use and risk of depression, psychotic symptoms, suicide, and attempted suicide. AB - BACKGROUND: It has been suggested that there is a causal association between isotretinoin therapy and the risk of depression, psychotic symptoms, suicide, and attempted suicide. OBJECTIVE: To further investigate the proposed association between isotretinoin therapy and the risk of depression, psychotic symptoms, suicide, and attempted suicide using a formal study design. DESIGN: Large population-based cohort studies. SETTING: The Canadian Saskatchewan Health Database and the United Kingdom General Practice Research Database. PATIENTS: Data were analyzed for 7195 isotretinoin users and 13,700 oral antibiotic users with acne from the Canadian Saskatchewan Health Database and for 340 isotretinoin users and 676 oral antibiotic users with acne from the United Kingdom General Practice Research Database. All subjects had computer-recorded histories of between 6 months and 5 years before, and at least 12 months after, their first isotretinoin or antibiotic prescription. OUTCOME MEASURE: Prevalence rates of neurotic and psychotic disorders, suicide, and attempted suicide were compared between isotretinoin and antibiotic users and within isotretinoin users as their own comparison (pretreatment vs posttreatment). The results were expressed as relative risks, calculated using multiple logistic regression analyses. RESULTS: Relative risk estimates, comparing isotretinoin use and oral antibiotic use with nonexposure to either drug for newly diagnosed depression or psychosis, were approximately 1.0 regardless of the data source. Similarly, relative risk estimates were all around 1.0 when comparing before with after isotretinoin use. The relative risk estimate for suicide and attempted suicide was 0.9 (95% confidence interval, 0.3-2.4) when comparing current isotretinoin exposure with nonexposure. CONCLUSION: This study provides no evidence that use of isotretinoin is associated with an increased risk for depression, suicide, or other psychiatric disorders. PMID- 11030770 TI - Isolated congenital nail dysplasia: a new autosomal dominant condition. AB - OBJECTIVE: Developmental nail abnormalities are extremely heterogeneous, with hereditary isolated conditions being a small and rare subgroup. An unusual congenital nail dysplasia observed in a large South German kindred was characterized clinically to review the question of uniqueness. DESIGN: Case series of affected family members. SETTING: University department of dermatology and houses of patients. PATIENTS: The history and clinical features in 22 affected family members (13 females and 9 males, aged 5 to 74 years) were recorded and documented by photographs. Nail biopsy samples were taken from 2 patients. INTERVENTIONS: None. RESULTS: The pedigree spanning 5 consecutive generations was best compatible with autosomal dominant inheritance with complete penetrance. Nail alterations were mostly present since birth and soon reached an individually variable degree of severity. Affected persons showed longitudinal streaks and thinning of nail plates, mostly of all fingernails and toenails, with some accentuation of the thumbnail and big toenails, poorly developed lunulae, longitudinal angular ridges of individual nail plates occasionally starting proximally from a reddish prominence, platonychia and koilonychia of individual nails often overgrowing the lateral folds, and notches and fissures of the free margins. Histological abnormalities included a prominent granular layer of the nail matrix and epithelial strands and buds extending from the nail bed. There were no associated anomalies. Other nail dystrophies were excluded by differences in clinical and histological features. CONCLUSION: The nail abnormality observed in our family represents a new autosomal dominant disorder for which we propose the term isolated congenital nail dysplasia. Arch Dermatol. 2000;136:1239-1243 PMID- 11030771 TI - Clinical and genetic studies of 3 large, consanguineous, Algerian families with Mal de Meleda. AB - BACKGROUND: Mal de Meleda (MIM 248300), also referred to as keratosis palmoplantaris transgrediens of Siemens, is a rare autosomal recessive skin disorder with a prevalence in the general population of 1 in 100,000. The main clinical characteristics are transgressive palmoplantar keratoderma, hyperhidrosis, and perioral erythema, but there are also associated features such as brachydactyly, nail abnormalities, and lichenoid plaques. OBSERVATIONS: We studied the clinical and genetic characteristics of 3 large, consanguineous, Algerian families, including 14 affected individuals. Homozygosity mapping of the third family confirmed localization of the responsible gene to 8qter in all 3 families. CONCLUSIONS: Although some differences in phenotypic expression among subjects were noted, genetic analysis of the 3 families who shared a common ethnic background indicated that a single gene is responsible for mal de Meleda in this population. PMID- 11030772 TI - Letter from Australia: life after the World Congress. PMID- 11030774 TI - Skin bioequivalents and their role in the treatment of inherited epidermolysis bullosa. PMID- 11030773 TI - Of mass and men: neurofibromas and histogenesis. PMID- 11030775 TI - Nonhealing leg ulceration. Diagnosis: protothecosis. PMID- 11030776 TI - Verrucous annular ulcerated hip plaques. Diagnosis: superficial granulomatous pyoderma form of pyoderma gangrenosum. PMID- 11030777 TI - Purplish papules on the legs. Diagnosis: type I cryoglobulinemia in association with WM. PMID- 11030778 TI - Asymptomatic plaque on the chin of a 10-year-old girl. Diagnosis: Striated muscle hamartoma. PMID- 11030779 TI - No evidence to support delay in excision of malignant melanoma. PMID- 11030780 TI - No evidence to support delay in excision of malignant melanoma PMID- 11030781 TI - Nicotine and pemphigus. PMID- 11030782 TI - Rigid definitions restrict the evolution of understanding. PMID- 11030783 TI - Rigid definitions restrict the evolution of understanding PMID- 11030784 TI - KH 1060 for the treatment of lichen planus: a multicenter, randomized, double blind, vehicle-controlled study. PMID- 11030785 TI - Livedo reticularis, ulcers, and peripheral gangrene: cutaneous manifestations of primary hyperoxaluria. PMID- 11030786 TI - Generalization of cicatricial pemphigoid during azathioprine therapy. PMID- 11030787 TI - Treatment of warts with Candida antigen injection. PMID- 11030788 TI - Association of Helicobacter pylori infection with psoriasis and lichen planus: prevalence and effect of eradication therapy. PMID- 11030789 TI - Randomized double-blind placebo-controlled trial in the treatment of alopecia areata with 0.25% desoximetasone cream. PMID- 11030790 TI - Lime disease? PMID- 11030791 TI - Is there a connection between the concentration of cholesterol circulating in plasma and the rate of neuritic plaque formation in Alzheimer disease? PMID- 11030792 TI - Imaging: window on the brain. AB - Given the rapid advances in neuroimaging and brain mapping, it is no surprise that a whole host of new and burgeoning techniques are on the horizon as we enter the 21st century. This brief review focuses on methods that are just entering the clinical experience or are now being explored in a research setting but have the imminent potential for clinical use. A number of brain mapping techniques now allow the clinician to monitor disease progression and therapeutic effects in either the routine clinical setting or experimental clinical trials. A battery of methods are now available for the preoperative and intraoperative evaluation of patients with lesions in or near critical cortical areas or for targeting purposes when deep nuclei of the brain are potential sites of therapeutic ablation or electrophysiologic stimulation. The development of probabilistic atlases will soon provide a means of understanding normal variants of human brain structure and function and studying brain disorders and their treatment in an objective and quantifiable fashion. Techniques that are now on the horizon for imaging gene expression, neuronal excitivity, and connectivity are presented in their current stage of development. It is clear that brain mapping and neuroimaging will continue to be ever more important parts of clinical neuroscience and may ultimately serve as the bridge between the molecular and clinical domains of this field. Arch Neurol. 2000;57:1413-1421 PMID- 11030793 TI - Homocysteine and neurologic disease. AB - Over the last 10 years, there has been an explosion of interest in homocysteine, a sulfur-containing amino acid that occupies a central location in the metabolic pathways of thiol compounds. This interest is primarily because of the realization that hyperhomocysteinemia is an important risk factor for vascular disease, including stroke, independent of long-recognized factors such as hyperlipidemia, hypertension, diabetes mellitus, and smoking. Since elevated homocysteine levels can often be normalized by supplementing the diet with folic acid (folate), pyridoxine hydrochloride (vitamin B(6)), and cyanocobalamin (vitamin B(12)), these observations raise the exciting possibility that this inexpensive and well-tolerated therapy may be effective in decreasing the incidence of vascular disease. In addition to its association with cerebrovascular disease, homocysteine may play a role in neurodegenerative disorders, even if only as a marker of functional vitamin B(12) deficiency. Homocysteine is also important to neurologists since most anticonvulsants raise homocysteine levels, an effect that may explain the teratogenic effects of these drugs. Practical knowledge concerning some details of homocysteine metabolism, the diagnosis of hyperhomocysteinemia, and the use of polyvitamin therapy to lower homocysteine levels will be increasingly important in the treatment of patients with neurologic disease. Arch Neurol. 2000;57:1422-1428 PMID- 11030794 TI - Thrombolytic therapy in patients with acute ischemic stroke. PMID- 11030795 TI - Decreased prevalence of Alzheimer disease associated with 3-hydroxy-3 methyglutaryl coenzyme A reductase inhibitors. AB - CONTEXT: Increasing evidence suggests that cholesterol plays a role in the pathophysiology of Alzheimer disease (AD). For instance, an elevated serum cholesterol level has been shown to be a risk factor for AD. OBJECTIVE: To determine whether patients taking 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins), which are a group of medicines that inhibit the synthesis of cholesterol, have a lower prevalence of probable AD. DESIGN: The experiment uses a cross-sectional analysis comparing the prevalence of probable AD in 3 groups of patients from hospital records: the entire population, patients receiving 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (hereafter referred to as the statins), and patients receiving medications used to treat hypertension or cardiovascular disease. PATIENTS: The subjects studied were those included in the computer databases of 3 different hospitals for the years October 1, 1996, through August 31, 1998. MAIN OUTCOME MEASURES: Diagnosis of probable AD. RESULTS: We find that the prevalence of probable AD in the cohort taking statins during the study interval is 60% to 73% (P < .001) lower than the total patient population or compared with patients taking other medications typically used in the treatment of hypertension or cardiovascular disease. CONCLUSIONS: There is a lower prevalence of diagnosed probable AD in patients taking 2 different 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors-lovastatin and pravastatin. While one cannot infer causative mechanisms based on these data, this study reveals an interesting association in the data, which warrants further study. Arch Neurol. 2000;57:1439-1443 PMID- 11030796 TI - Postoperative changes in cerebral metabolism in temporal lobe epilepsy. AB - BACKGROUND: Fludeoxyglucose F 18 positron emission tomography ((18)F-FDG-PET) can detect focal metabolic abnormalities ipsilateral to the seizure focus in 80% of patients with temporal lobe epilepsy (TLE). Regions outside the epileptogenic zone can also be affected. We hypothesized that these remote regions might show altered metabolism, tending to return toward normal values, after surgery. DESIGN: Interictal preoperative and postoperative (18)F-FDG-PET metabolism were compared in patients with refractory TLE. Based on pathological findings, disease was classified in the following 3 groups: mesial temporal sclerosis, mass lesions, and no pathological diagnosis. Quantitative PET data analysis was performed using the region-of-interest template previously described. Global normalization was used to adjust for the effect of antiepileptic medication changes. Data were analyzed by Wilcoxon signed rank test and analysis of variance. SETTING: The Clinical Epilepsy Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health. PATIENTS: Twenty-two patients with refractory TLE. RESULTS: Preoperatively, in all groups, cerebral metabolic rate for glucose was decreased ipsilateral to the resection site in inferior lateral temporal, inferior mesial temporal, and inferior frontal areas and thalamus. Postoperatively, in all groups, cerebral metabolic rate for glucose increased in ipsilateral inferior frontal area and thalamus. In the mesial temporal sclerosis group, we found a statistically significant increase in the contralateral thalamus. CONCLUSION: Temporal lobe epilepsy is associated with extensive preoperative decreased metabolism in inferior lateral temporal, inferior mesial temporal, and inferior frontal areas and thalamus. Postoperatively, we found increased IF and thalamic metabolism. Seizures may have a reversible effect on brain areas connected with, but remote from, the epileptogenic cortex. Arch Neurol. 2000;57:1447-1452 PMID- 11030797 TI - Novel presenilin 1 mutations associated with early onset of dementia in a family with both early-onset and late-onset Alzheimer disease. AB - Two children of an adult with early-onset, autopsy-confirmed Alzheimer disease (AD) developed dementia in their late 20s and were subsequently found to have novel mutations in codon 434 of the presenilin 1 (PS1) gene on chromosome 14, a G to-T substitution at nucleotide 1548 and a C-to-G substitution at nucleotide 1549. The younger of the 2 children had AD confirmed at postmortem examination. The disease course in these 3 individuals was characterized by cognitive and behavioral problems accompanied by myoclonus, seizures, and aphasia within 5 years after onset. Two grandparents had clinically diagnosed AD with stroke beginning at ages 78 and 66 years, but neither had a PS1 mutation. No other living family member was demented, nor did any other family member have the PS1 mutation. We conclude that the affected parent of the proband was a likely recent founder for these novel mutations in PS1. The family demonstrates the clinical and genetic heterogeneity of AD. Arch Neurol. 2000;57:1454-1457 PMID- 11030798 TI - Acute orthostatic hypotension when starting dopamine agonists in Parkinson's disease. AB - OBJECTIVE: To study the frequency and severity of acute orthostatic hypotension (OH) in patients with Parkinson's disease who are starting dopamine agonist therapy. PATIENTS AND METHODS: In the context of an outpatient clinical practice, 29 consecutive patients with Parkinson's disease who were starting dopamine agonist therapy were brought into the clinic for their first dose of agonist. After a baseline supine and standing blood pressure assessment, patients were given a test dose of either pergolide mesylate (0.025, 0.05, 0. 125, or 0.25 mg), pramipexole dihydrochloride (0.125 mg), or ropinirole hydrochloride (0.125 or 0.25 mg). At 3 selected times, blood pressure readings were repeated in the supine and standing positions. MAIN OUTCOME MEASURE: Orthostatic hypotension was defined as a drop in either systolic blood pressure of more than 25 mm Hg or diastolic pressure of more than 10 mm Hg. Patients with OH before the administration of the dopamine agonist were excluded. RESULTS: Ten subjects (34%) met the criteria for acute OH. There was no evidence that OH was related to the use of a specific dopamine agonist or the concurrent use of levodopa. Of the patients who met the criteria for OH, only 3 (30%) had symptoms of OH, such as lightheadedness or general malaise. CONCLUSIONS: Acute OH occurs frequently when starting dopamine agonist therapy in Parkinson's disease, but is frequently not appreciated by patients. Knowledge of acute blood pressure responses may be useful when making decisions regarding agonist titration schedules in clinical practice. Arch Neurol. 2000;57:1461-1463 PMID- 11030799 TI - Computerized posturography analysis of progressive supranuclear palsy: a case control comparison with Parkinson's disease and healthy controls. AB - BACKGROUND: Progressive supranuclear palsy (PSP) is a neurodegenerative disorder that is frequently mistaken for Parkinson's disease (PD) in its early stages. OBJECTIVE: To compare balance measures using computerized posturography in patients with early PSP and early PD. METHODS: We performed computerized posturography (SMART Balance Master; NeuroCom International, Inc, Clackamas, Ore) in 20 patients with clinically diagnosed mild to moderate PSP (ambulatory) and compared results with those from 20 patients with PD of similar age and disease duration who were not receiving medications, and from 20 healthy age- and sex matched controls. Sensory organization testing (SOT), limits of stability (LOS), and toes-up perturbations (4 degrees at 50 degrees per second) were tested while receiving and not receiving a combination of oral carbidopa (25 mg) and levodopa (250 mg) in the PSP group. Clinical assessment included Unified Parkinson's Disease Rating Scale, Performance-oriented assessments, and functional reach. RESULTS: When compared with the PD and control groups, total LOS time (P < .001) and path sway (P < .001) were significantly prolonged in PSP. Total SOT showed significantly worse scores in PSP compared with PD and control groups (F(2,57) = 29.6; P < .001). Univariate follow-up tests comparing PSP and PD showed differences in the following conditions: eyes open and visual sway (P = .003), eyes open and platform sway (P = .003), eyes closed and platform sway (P < .001), and eyes open and platform and visual sway (P < .001). Medium- and long-latency responses to perturbation were similar, but a larger number in the PSP group lacked short-latency responses (chi(2) = 11.3; P = .002). Levodopa administration did not significantly improve any aspect of posturography testing in PSP. In differentiating PSP from PD, LOS time and SOT condition of eyes open and platform and visual sway were nearly 100% sensitive and 100% specific (canonical correlation, 0.91). CONCLUSIONS: Computerized posturography testing reliably differentiated early PSP from early PD and age-matched controls. The PSP group demonstrated severely contracted limits of stability with probable deficits in motor programming. Results of SOT in PSP suggested a vestibular pattern and overreliance on visual cues, even when incorrect. The absence of short-latency responses (monosynaptic reflex arch) suggests an additional disturbance in the spinal cord or peripheral nervous system. Arch Neurol. 2000;57:1464-1469 PMID- 11030800 TI - Small concomitant vascular lesions do not influence rates of cognitive decline in patients with Alzheimer disease. AB - OBJECTIVE: To determine the relation between concomitant small cerebral infarction and clinical progression of Alzheimer disease (AD). DESIGN: A retrospective clinicopathologic study of patients with AD. METHODS: We searched the databases of the University of California, San Diego, Alzheimer's Disease Research Center, La Jolla, for patients with an autopsy diagnosis of definite AD with or without a concomitant small cerebral infarction. Clinical and neuropsychologic data obtained during longitudinal follow-up were available for 201 subjects with AD neuropathologic features and 36 with AD and concomitant cerebral infarcts (volume, < 10 cm(3)). The rates of cognitive decline on the Mini-Mental State Examination and the Dementia Rating Scale were each calculated and compared between the 2 groups. RESULTS: The age at death was significantly (P = .05) higher and the Braak stage was lower in patients with mixed AD and infarct pathological features compared with those with AD pathological features only. The rate of cognitive decline over time was not significantly (P > or = .20 for all) different between the 2 groups. There was a trend for the presence of a cerebral infarct to be associated with more severe clinical dementia (P =.08) as measured by the Dementia Rating Scale, but no such trend for the Mini-Mental State Examination. CONCLUSION: This clinicopathologic correlation study suggests that concomitant small cerebral infarcts with a total volume of less than 10 cm(3) do not significantly influence the overall rate of global cognitive decline in patients with AD. Arch Neurol. 2000;57:1474-1479 PMID- 11030801 TI - Prospective study of apolipoprotein E genotype and functional outcome following ischemic stroke. AB - BACKGROUND: The apolipoprotein E (APOE) epsilon 4 allele is a marker of adverse outcome following head injury and intracerebral hemorrhage. Transgenic animal data in a focal cerebral ischemia model suggest that the epsilon 4 allele increases infarct size and functional impairment. OBJECTIVE: To determine if APOE genotype is associated with functional recovery from ischemic stroke. DESIGN: Prospective study. SETTING: Stroke service at a university teaching hospital. PATIENTS: Patients with clinical and neuroimaging findings (computed tomography or magnetic resonance imaging) compatible with an acute ischemic stroke. MAIN OUTCOME: Functional outcome by Barthel index (BI) and modified Rankin scale (mRS) was compared for epsilon 3/epsilon 3 patients vs epsilon 4 carriers and vs epsilon 2 carriers at 1 and 3 months. Univariate predictors of 3-month outcome were examined in a multivariate analysis. RESULTS: One hundred eighty nine patients were enrolled: 100 women, 89 men (mean +/- SD age, 69.4 +/- 11.0 years). There were 25 epsilon 2 alleles (frequency, 0.07), 292 epsilon 3 alleles (0.77), and 61 epsilon 4 alleles (0.16). Baseline National Institutes of Health Stroke Scale scores and Oxfordshire Community Stroke Project classifications were similar in all groups (epsilon 3/epsilon 3, epsilon 4, and epsilon 2 carriers). One-month (BI, P = .64; mRS, P = .59) and 3-month (BI, P = .87; mRS, P = .73) outcomes were not associated with possession of either epsilon 4 or the epsilon 2 allele. Baseline National Institutes of Health Stroke Scale scores (P < .001) and age (P = .002) were significant predictors of 3-month BI and mRS outcomes in multivariate analyses. CONCLUSIONS: Although there is a robust influence of APOE polymorphism on functional recovery after some types of brain injury in humans, it does not exert a major influence on injury severity or functional recovery following ischemic stroke. Arch Neurol. 2000;57:1480-1484 PMID- 11030803 TI - Extrapyramidal motor signs in degenerative ataxias. AB - BACKGROUND: Extrapyramidal motor signs (EPS) are well-known symptoms of degenerative ataxia. However, little is known about frequency and appearance of EPS in subtypes of ataxia. METHODS: We characterized 311 patients with ataxia clinically and genetically. Course of the disease and EPS were investigated according to a standardized protocol. Diagnostic and prognostic impact of EPS in subtypes of ataxia was analyzed by Kaplan-Meier plots. RESULTS: Extrapyramidal motor signs occurred in all forms of ataxia, but frequency and type of EPS varied between genetically and clinically defined subtypes. Postural tremor in hereditary ataxias was typical for spinocerebellar ataxia type 2 (SCA2). Dystonia was generally rare in ataxias, but, if present, suggested SCA3. We observed a parkinsonian variant of SCA3 in which parkinsonism was present in the beginning of the disease and responded well to levodopa therapy, leading to diagnostic confusion. Parkinsonism in SCA3 was independent of CAG repeat length but ran in families, suggesting modifying genes. In idiopathic sporadic cerebellar ataxia (ISCA), EPS are more frequent in late-onset than in early-onset forms. In 50% of ISCA patients with parkinsonism, the diagnosis of multiple system atrophy remained questionable because of normal autonomic function. CONCLUSIONS: Extrapyramidal motor signs can help to predict the genetic subtype of ataxia. Extrapyramidal motor signs were more frequent in genetic subtypes in which basal ganglia affection has been demonstrated by postmortem studies. However, no type of EPS was specific for an underlying mutation. In ISCA, EPS are an adverse prognostic factor. Parkinsonism is especially associated with a more rapid course of the disease. Arch Neurol. 2000;57:1495-1500 PMID- 11030802 TI - Quality of life and its relationship to brain lesions and atrophy on magnetic resonance images in 60 patients with multiple sclerosis. AB - CONTEXT: Disease-modifying multiple sclerosis (MS) therapeutic trials continue to rely on physical disability as the main clinical outcome measure, while the impact of treatment on quality of life (QOL) is poorly understood. Weak correlations exist between physical disability and the disease burden as shown using conventional brain magnetic resonance imaging (MRI), indicating poor sensitivities of these measures alone in defining the clinical course of MS. OBJECTIVES: To investigate the impact of MS on QOL; to determine whether impaired QOL in patients with MS was related to any regional brain abnormalities assessed using conventional MRI sequences; and to determine if the severity of MS as assessed by the Expanded Disability Status Scale (EDSS) and clinical course was associated with worsening QOL. DESIGN, SETTING, AND PATIENTS: Prospective, cross sectional study of 60 consecutive patients with MS treated in a community-based, university-affiliated MS clinic. MAIN OUTCOME MEASURES: Assessments of QOL using the Multiple Sclerosis Quality of Life-54 Instrument were correlated with the scores of the EDSS, clinical course, and findings on brain MRI. RESULTS: Quality of life was significantly impaired in patients with MS and was worse in patients with secondary-progressive MS compared with those with relapsing-remitting MS. Brain MRI lesions and atrophy were associated with impaired QOL with respect to sexual dysfunction, overall mental health, and limitations due to physical and emotional dysfunction. Correlations between MRI results and QOL assessments were much stronger for hypointense lesions and atrophy on T1-weighted images than for hyperintense lesions on T2-weighted images and were insignificant for lesions on contrast-enhanced images. Higher EDSS scores were associated with impairments in most physical and mental health QOL scales but were weakly correlated with cognitive and sexual dysfunction. CONCLUSIONS: In patients with MS, QOL is impaired and is associated with increasing neurologic disability. Quality of life assessments are related in part to brain lesions and atrophy shown on MRI. Assessments of QOL provide unique information not readily evaluated by EDSS and may be useful as secondary clinical outcome measures. Arch Neurol. 2000;57:1485 1491 PMID- 11030804 TI - Vitamin E and beta carotene supplementation in high risk for stroke: a subgroup analysis of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study. AB - CONTEXT: High serum or dietary levels of vitamin E and beta carotene appear to be associated with lower risk of stroke, but studies regarding their supplementation have not supported their use in stroke prevention. OBJECTIVE: To determine if vitamin E (dl-alpha tocopherol) and beta carotene supplementations could be used in prevention of stroke in men at high risk for hemorrhagic or ischemic events. DESIGN: Population-based, randomized, double-blind, placebo-controlled, 2 x 2 factorial design trial (the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study), conducted from April 1985 through April 30, 1993, with median follow-up of 6 years. INTERVENTIONS: Alpha tocopherol, 50 mg; beta carotene, 20 mg; both; or placebo. PARTICIPANTS: From the total male population aged 50 through 69 years in southwestern Finland (n = 290,406), 29,133 male smokers were randomized to 1 of 4 treatment regimens. We excluded 614 men because of previous stroke at baseline, leaving 28, 519. MAIN OUTCOME MEASURES: Incident and fatal subarachnoid and intracerebral hemorrhage, cerebral infarction, and unspecified stroke. RESULTS: Stroke occurred in a total of 1057 men: 85 had subarachnoid and 112 had intracerebral hemorrhage, 807 had cerebral infarction, and 53 had unspecified stroke. Within 90 days from onset, 160 men died of stroke. Vitamin E supplementation increased the risk of subarachnoid hemorrhage (relative risk [RR], 2.45; 95% confidence interval [CI], 1.08-5.55) and decreased risk of cerebral infarction (RR, 0.70; 95% CI, 0.55-0.89) in hypertensive men but had no effect among normotensive men. Furthermore, it decreased the risk of cerebral infarction, without elevating the risk of subarachnoid hemorrhage, among hypertensive men with concurrent diabetes (RR, 0.33; 95% CI, 0.14-0.78). Beta carotene supplementation appeared to increase the risk of intracerebral hemorrhage and modestly decrease that of cerebral infarction among men with greater alcohol consumption. CONCLUSION: Vitamin E supplementation may prevent ischemic stroke in high-risk hypertensive patients, but further studies are needed. Arch Neurol. 2000;57:1503-1509 PMID- 11030805 TI - Hemidystonia and hemichoreoathetosis as an initial manifestation of moyamoya disease. AB - OBJECTIVE: To describe hemidystonia and hemichoreoathetosis in an adult patient with moyamoya disease without a previous history of cerebrovascular accident. DESIGN: Case report. SETTING: Tertiary care center. PATIENT: A 22-year-old woman suddenly developed dystonic spasms in her left hand and left foot after a severe emotional stress. The dyskinesia gradually subsided over the next 4 months. Five months after the onset, she suddenly developed choreoathetoid movement in her right hand and right foot. MAIN OUTCOME AND RESULTS: The patient had both somatic and cortical sensory deficits in the right hand and right foot. Magnetic resonance imaging of the brain showed an infarction at the right putamen and lesions involving the right frontal lobe and the left frontotemporoparietal lobe. Magnetic resonance cerebral angiography showed severe stenoses of both internal carotid arteries at the supraclinoid portion and numerous collateral vessels, compatible with moyamoya disease. Single photon emission tomography of the brain showed hypoperfused areas at the right frontal and left frontotemporoparietal lobes. The choreoathetosis of the right limbs improved markedly, along with improvement of sensory deficits. CONCLUSIONS: To our knowledge, this is the first report of an adult patient presenting with hemidystonia and hemichoreoathetosis as the initial manifestations of moyamoya disease. Arch Neurol. 2000;57:1510-1512 PMID- 11030807 TI - Pramipexole is a possible effective treatment for primary orthostatic tremor (shaky leg syndrome). AB - OBJECTIVE: To report the successful treatment of primary orthostatic tremor in a 76-year-old man. BACKGROUND: Primary orthostatic tremor is a rare condition, with few reports describing therapies. Established therapies had previously failed in our patient. METHODS: Using an evidence-based evaluation of treatments via MEDLINE's GRATEFUL MED search engine, a therapeutic option was determined during the first consultation with the patient, and pramipexole therapy was initiated. RESULTS: The therapy proved effective, and the patient had relief from his symptoms for the first time in 6 years. CONCLUSION: Pramipexole is a potential therapy for primary orthostatic tremor. Arch Neurol. 2000;57:1519-1520 PMID- 11030806 TI - Molecular and clinical study of spinocerebellar ataxia type 7 in Chinese kindreds. AB - OBJECTIVE: To investigate the clinical and molecular characteristics of spinocerebellar ataxia type 7 (SCA7) in Chinese kindreds. BACKGROUND: Spinocerebellar ataxia type 7 is caused by the expansion of an unstable CAG repeat in the first exon of the SCA7 gene. METHODS: Clinical and related examinations were performed in all affected or at-risk individuals from 4 Chinese families presenting with autosomal dominant cerebellar ataxia and decreased visual acuity. The size of the (CAG)(n) array of the SCA7 gene was detected by polymerase chain reaction, polyacrylamide gel electrophoresis, and related techniques in the 4 families and 67 healthy controls. The relationship between expanded repeat number and age of onset was statistically analyzed. RESULTS: The SCA7 mutation was identified in 2 families. Clinical study revealed that great variation occurred in the age of onset, initial symptoms, and associated signs. Meanwhile, the analysis of 11 parent-child couples demonstrated the existence of marked anticipation. Some distinct retinal changes were noted in 2 affected patients. All SCA7 patients in our series exhibited expanded CAG repeats, ranging from 44 to 85 repeats, with a strong negative correlation between repeat size and age of onset. Repeat lengths of expanded alleles showed somatic mosaicism in leukocyte DNA. There were some subtle clinical differences between the SCA7 positive and -negative cases. CONCLUSIONS: Clinical variation occurred not only among the SCA7 families but also within the same kindred. Meiotic and mitotic instability of the CAG repeat in the SCA7 gene were demonstrated, and intergenerational instability of the array was associated with the clinical phenomenon of anticipation. Arch Neurol. 2000;57:1513-1518 PMID- 11030808 TI - Retropharyngeal abscess. PMID- 11030809 TI - Essential tremor. PMID- 11030810 TI - Cerebellar ataxia. PMID- 11030811 TI - Weighing the neurological complexities of long-term levodopa use. PMID- 11030813 TI - Ultrasound biomicroscopy of the eye in cystinosis. AB - OBJECTIVE: To describe the ocular ultrasound biomicroscopy (UBM) findings in patients with cystinosis. METHODS: Six patients with infantile nephropathic cystinosis, aged 16 to 25 years, and 6 controls (matched for age and spherical refractive error) were examined clinically and with UBM. Scleral reflectivity, corneal and iris thickness, central anterior chamber depth, angle width, trabecular meshwork to ciliary process distance, and ciliary sulcus width were measured. RESULTS: No patient had glaucoma or posterior synechiae, but all had crystals in the trabecular meshwork apparent with gonioscopy. Using UBM, the cornea and iris appeared similar in both groups, but the scleral reflectivity was increased in patients (P =.003). The angle was narrower in patients (mean +/- SD, 20 degrees +/- 7 degrees ) than controls (31 degrees +/- 5 degrees, P<. 001). The anterior chamber was shallower in patients (2556 +/- 197 microm) than controls (2968 +/- 284 microm, P<.001). The ciliary sulcus was closed or narrow in all patients (83 +/- 112 microm) compared with controls (339 +/- 135 microm, P<.001), with a reduction in the trabecular meshwork to ciliary process distance. CONCLUSIONS: This report of ocular UBM findings in cystinosis demonstrated narrowing of the angle and a ciliary body configuration similar to that reported for plateau iris syndrome. Gonioscopy demonstrated crystals in the trabecular meshwork. These findings may explain the predisposition of these patients to glaucoma. PMID- 11030814 TI - Long-term effect of Nd:YAG laser posterior capsulotomy on intraocular pressure. AB - OBJECTIVE: To study the long-term effect of Nd:YAG capsulotomy on intraocular pressure (IOP). METHODS: We reviewed the records of patients with bilateral pseudophakia who received Nd:YAG capsulotomy in only 1 eye. Using the Wilcoxon rank sum test, we compared the mean change in IOP in eyes before and after capsulotomy with that of the noncapsulotomy eyes at corresponding time intervals. Using multiple regression tests, we analyzed the factors significantly associated with postcapsulotomy long-term IOP increases. RESULTS: The study included 100 patients who were followed up for a median of 1.5 years after capsulotomy. The mean +/- SD age of the study group was 76+/-7 years, and 37 patients had glaucoma. The changes in IOP in the eyes treated with capsulotomy were significantly higher than those in noncapsulotomy eyes at each time interval following capsulotomy. The long-term IOP increase was significantly associated with the IOP increase measured 1 hour after the capsulotomy (P =. 001). Patients with glaucoma were more likely to require long-term additional glaucoma medication than were nonglaucoma patients to require initial glaucoma therapy after the capsulotomy (P =.002). CONCLUSION: After Nd:YAG capsulotomy, long-term IOP is often elevated above precapsulotomy baselines, especially in glaucoma patients or patients who experience a significant IOP increase within hours after the capsulotomy. PMID- 11030815 TI - Postoperative application of mitomycin for trabeculectomies. AB - BACKGROUND: Persistent hypotony is a severe complication following trabeculectomy with intraoperative application of mitomycin. OBJECTIVE: To reduce this rate of hypotony by using a lower concentration of mitomycin and applying the mitomycin only topically to the filtering bleb following surgery. METHODS: Patients were enrolled on a consecutive basis and prospectively followed up. Standard trabeculectomies were performed and mitomycin applied postoperatively on the 3 days following surgery (group 1). For comparison, data from previous studies were used for control eyes with intraoperative mitomycin application (group 2) and no mitomycin application (group 3). Preoperative and postoperative data, complications, and the need for further surgical procedures were evaluated. RESULTS: The study group (group 1) consisted of 22 cases. The mean follow-up was 13.4 and 13.5 months for groups 1 and 2, respectively. Average intraocular pressure values decreased from 33. 6 and 31.0 mm Hg (P =.32; t test) to 16.0 and 12.5 mm Hg in the 2 groups (P =.03; t test). The average number of medications decreased from 2.5 and 2.5 to 0.6 and 0.4 (P =.35; t test) in groups 1 and 2, respectively, at the last visit. Hypotony lasting for more than 3 months occurred only in eyes with intraoperative mitomycin application (14/22). Choroidal detachment (3/22) and hypotony maculopathy (2/22) also were only noted in eyes from group 2. In group 3, success rates were much lower. CONCLUSIONS: This is the first clinical study, to our knowledge, to evaluate the efficacy of this new technique of mitomycin application. From the results, it appears that the postoperative application of mitomycin following trabeculectomy is associated with a lower risk of severe and long-standing hypotony. This technique may be promising in eyes at low risk for failure. PMID- 11030816 TI - Transplantation of autologous iris pigment epithelium after removal of choroidal neovascular membranes. AB - BACKGROUND: Transplantation of autologous iris pigment epithelium (IPE) into the subretinal space has been suggested as one approach for the treatment of age related macular degeneration, as well as for other conditions in which loss of retinal pigment epithelium (RPE) occurs. Surgical removal of choroidal neovascular membranes is associated with traumatic loss of the RPE cell layer, disruption of the integrity of the photoreceptor-RPE complex, and limited visual outcome. OBJECTIVE: To examine whether IPE cells can substitute for RPE cells to be transplanted to the subretinal space of patients with either RPE degenerative disease or traumatic loss of the RPE cell layer after subretinal surgery. METHODS: Autologous IPE cells were transplanted to the subretinal space in 20 consecutive patients undergoing removal of subretinal fibrovascular membranes using pars plana vitrectomy. Autologous IPE cells were harvested by iridectomy, isolated, and transplanted directly to the subretinal spaces. Transplants were evaluated for 6 to 11 months by funduscopy, fluorescein angiography, and scanning laser ophthalmoscopic (SLO) microperimetry. RESULTS: For the entire follow-up period, no evidence of any immunologic response was observed. Revisional surgery was necessary in 3 patients because of complications (rhegmatogenous retinal detachment [n = 1]; proliferative vitreoretinopathy [n = 1]; and macular pucker [n = 1]); 1 patient did not receive IPE cells. Five of 19 phakic eyes underwent cataract surgery; in 1 case this was combined with the vitrectomy. Five patients showed improved visual acuity of 3 to 4 lines, 13 patients had stable visual acuity (+/-2 lines), and 2 patients had reduced visual acuity of 6 lines. CONCLUSIONS: In this pilot study, the transplantation of autologous IPE cells was done as an addition to conventional surgical excision of choroidal neovascular membranes. Transplanted cells were well tolerated in the subretinal space and did not adversely affect the function of the photoreceptors, since improvement or stable visual acuity was observed in 18 patients after IPE transplantation. These results suggest that autologous IPE cells may be used as a substitute for autologous RPE cells to transplant to the subretinal space to treat age-related macular degeneration. PMID- 11030817 TI - Pattern of vascular nonperfusion in retinal venous occlusions occurring within the optic nerve with and without optic nerve head swelling. AB - OBJECTIVE: To investigate the significance of optic nerve head swelling (ONHS) in relation to the pattern of vascular nonperfusion, visual acuity (VA), and demographic profile in retinal venous occlusions (RVOs) occurring within the optic nerve. METHODS: Cases of RVO occurring within the optic nerve were divided on the basis of the presence (105 cases) or absence (163 cases) of ONHS. This division was performed by examining the color stereo fundus photographs in conditions masked from other clinical parameters. Duration of symptoms before assessment, age, and sex distributions were compared. The vein involved was identified, and the occlusion was confirmed to have occurred within the optic nerve by observing that the vein pierced the lamina cribrosa as a dilated vein. Fluorescein angiographs were examined, and the extent of vascular nonperfusion in the macula and peripheral retina was quantified from grade 1 to grade 4. The extent of break in the perifoveal capillary arcade was graded as 0, less than or equal to 90 degrees, and greater than 90 degrees. Best-corrected VA was assessed using the Snellen chart. RESULTS: The 2 groups were comparable in terms of the duration of the symptoms before examination. The mean age was significantly younger in the group with ONHS (58.3 vs 65.1 years, P<. 001). Age distribution by sex demonstrated a higher proportion of men younger than 50 years in the ONHS group (19.1% vs 8.6%, P =.01). The group without ONHS involved the papillary vein more frequently (31.3% vs 17.1%, P =.01). The respective proportions of grade 1, 2, 3, and 4 vascular nonperfusion in the macula were 90.5%, 9.5%, 0%, and 0% in the ONHS group, and 62.6%, 14.7%, 13.5%, and 9.2% in the group without ONHS (P<.001). The corresponding proportions for the peripheral retina were 90.4%, 8.7%, 0%, and 1.0% in the ONHS group, and 62.7%, 13.0%, 18.0%, and 6.2% in the group without ONHS (P<.001). In 64.6% of cases with ONHS and 42.9% of cases without, the perifoveal arcade was intact. A break greater than 90 degrees in the perifoveal arcade was present in 12.5% of cases with and 23.6% of cases without ONHS (P =.004). The median VA was significantly better in the ONHS group (6/24 vs 6/48, P =.005). CONCLUSIONS: The RVOs occurring within the optic nerve can be subdivided into 2 distinct groups on the basis of ONHS. The presence of ONHS is associated with younger age, less severe vascular nonperfusion, and better VA. This is consistent with a retrocribrosal site of occlusion, which has access to the pial plexus that can provide collateral channels for retinal venous drainage. PMID- 11030818 TI - Alterations of the blood-retinal barrier and retinal thickness in preclinical retinopathy in subjects with type 2 diabetes. AB - OBJECTIVE: To identify alterations of the blood-retinal barrier by mapping retinal fluorescein leakage into the vitreous and changes in retinal thickness occurring in the macular region in preclinical diabetic retinopathy. METHODS: Ten eyes from 10 patients with type 2 diabetes and no lesions visible on fundus photography (level 10 of Wisconsin grading) were examined with the retinal leakage analyzer (RLA) (Confocal Scanning Laser Ophthalmoscope [modified]; Carl Zeiss Inc, Thornwood, NY) and the retinal thickness analyzer (RTA) (Talia Technology, Mevaseret Zion, Israel). The maps of retinal leakage and retinal thickness were aligned and integrated in the same image to correlate leakage with thickness. Data from the group of individuals with diabetes were compared with those of a healthy control population (N = 14; mean age, 48 years; range, 42-55 years) and used to establish reference maps for the RLA and RTA. RESULTS: Areas of abnormally increased fluorescein leakage were detected in 9 of 10 eyes examined. The increased leakage in 6 (67%) of 9 eyes reached values higher than 40% more than the mean +2 SD RLA control value. Areas of abnormally increased thickness were found in 7 of 10 eyes examined. For the most part, the increases in retinal thickness were not severe (ie, <15% increase in 5 eyes and an 18% increase in 1 eye). The eyes with the most extensive leakage (cases 1, 3, and 9) showed relatively good coincidence between the location of the areas of increased leakage and the location of the areas of increased thickness. In 4 eyes (cases 2, 5, 7, and 8), no such correlation was apparent. The 3 remaining eyes showed little coincidence between these locations. Characteristically, the latter 3 eyes had areas of abnormally increased thickness that were much larger than the areas of increased fluorescein leakage, which were relatively moderate or absent of any leakage. CONCLUSIONS: Localized sites of increased fluorescein leakage and zones of increased retinal thickness were found in most eyes in a series of 10 eyes in the preretinopathy stage from 10 patients with type 2 diabetes. Increases in retinal thickness may be observed that do not coincide with sites of retinal leakage. Two types of increased retinal thickness may, therefore, be present in the preretinopathy stage of diabetic retinopathy, one directly associated with an alteration of the blood-retinal barrier, and another occurring without apparent breakdown of blood-retinal barrier. PMID- 11030819 TI - Visual outcome after surgical removal of choroidal neovascularization in pediatric patients. AB - OBJECTIVE: To assess the visual outcome after surgical removal of choroidal neovascularization (CNV) in pediatric patients. METHODS: A retrospective, noncomparative, consecutive case series of 17 eyes undergoing surgical removal of CNV of various causes in patients aged 18 years and younger. RESULTS: The cause of the CNV was presumed ocular histoplasmosis syndrome in 11 eyes, idiopathic in 3, and optic nerve coloboma, ocular toxoplasmosis, and trauma in 1 eye each. Two eyes had CNV within 100 microm of the center of the foveal avascular zone (juxtafoveal CNV) and 1 eye had peripapillary CNV, while 14 eyes had CNV beneath the geometric center of the foveal avascular zone (subfoveal CNV). In eyes with subfoveal CNV, median preoperative Snellen visual acuity was 20/200 (range, 20/80 3/200). With a median follow-up of 27 months (range, 6-45 months), median final visual acuity was 20/50 (range, 20/20-2/200); 10 (72%) had improvement of 2 or more Snellen lines after surgery, and 6 eyes (43%) had final visual acuity of 20/40 or better. In 3 eyes with juxtafoveal CNV or peripapillary CNV, all eyes had improvement of 4 or more Snellen lines. Postoperative recurrent CNV developed in 6 (35%) of 17 patients; 2 of the eyes underwent a second vitrectomy and 4 received laser treatment for the recurrences. CONCLUSIONS: Pediatric patients may have good recovery of vision after surgical removal of CNV, and the removal of these membranes may be a viable alternative to laser photocoagulation in pediatric patients. PMID- 11030820 TI - Frequency of the common canaliculus: a radiological study. AB - BACKGROUND: It is frequently stated in the literature that there is a common canaliculus (CC) in the lacrimal drainage system in about 90% of individuals; in the remaining 10%, the upper and lower canaliculi enter the lacrimal sac separately. To our knowledge, there is no quantitative study supporting this assumption. OBJECTIVE: To investigate the frequency of the CC and other anatomical types connecting the upper and lower canaliculi to the lacrimal sac. MATERIALS AND METHODS: We presumed that there might be 3 different types of anatomical connection between the lacrimal sac and the canaliculi. Digital subtraction macrodacryocystograms obtained from 281 patients with obstructive epiphora were reviewed, and those of 247 patients were included in the study. These patients were grouped according to the anatomical types. RESULTS: Dacryocystographies were performed on 153 patients unilaterally and 94 bilaterally. We observed a CC in 321 (94.1%) of 341 lacrimal drainage systems. In 13 lacrimal drainage systems (3.8%), the CC was absent, but the upper and lower canaliculi joined at the wall of the lacrimal sac. In only 7 lacrimal drainage systems (2.0%) did the upper and lower canaliculi enter the sac separately. In 2 patients with bilateral dacryocystograms, a CC was seen on one side but not on the other side. CONCLUSIONS: The frequency of separate drainage of the upper and lower canaliculi into the lacrimal sac is only 2.0%. The CC might not be present bilaterally in all individuals. PMID- 11030821 TI - Chronic infantile neurological cutaneous and articular/neonatal onset multisystem inflammatory disease syndrome: ocular manifestations in a recently recognized chronic inflammatory disease of childhood. AB - OBJECTIVE: To report on the ocular manifestations of the Chronic Infantile Neurological Cutaneous and Articular/Neonatal Onset Multisystem Inflammatory Disease (CINCA/NOMID) syndrome, a rare, recently identified, pediatric multisystem inflammatory disease with chronic cutaneous, neurological, and articular manifestations. DESIGN: Descriptive case-report study. SETTING: International collaborative study based on a questionnaire. RESULTS: We included 31 patients. The mean age at onset of eye manifestations was 4.5 years. Optic disc changes were the most common feature, occurring in 26 patients (83%), including optic disc edema, pseudopapilledema, and optic atrophy. Anterior segment manifestations varying from mild to severe were seen in 13 patients (42%); chronic anterior uveitis, in 17 patients (55%). Moderate to severe visual acuity loss in at least 1 eye was seen in 8 patients (26%) as a consequence of the disease. Posterior synechia, glaucoma, and white iritis were not observed in any patient. CONCLUSION: Ocular manifestations with potentially sight-threatening complications occur commonly in the CINCA/NOMID syndrome. The distinctive nature of these complications may assist the ophthalmologist in recognizing this rare disorder and distinguishing it from juvenile rheumatoid arthritis. PMID- 11030822 TI - Selective loss of S-cones in diabetic retinopathy. AB - OBJECTIVE: To determine whether selective cone loss could explain the acquired tritan-like color confusion found in diabetic retinopathy. METHODS: Terminal deoxynucleotidyl transferase-mediated biotin-deoxyuridine triphosphate nick end labeling (TUNEL) was employed on paraffin sections of retinas from 5 donors with diabetic retinopathy. For quantitative analysis, postmortem retinas were obtained from 13 human donors; 7 from patients with various durations and stages of diabetic retinopathy (4 background, 3 proliferative) and 6 controls. Enzyme histochemical analysis for carbonic anhydrase (CA) was used to distinguish L/M cones (positive for CA) from S-cones (negative for CA). Cone topography was determined by sampling 360 degrees from 0.1 to 1.5 mm of foveal eccentricity and along the horizontal meridians from 1.5 to 15.0 mm. RESULTS: Rare cells in both the inner and outer nuclear layers of the diabetic eyes were positively labeled with the TUNEL method. The CA staining revealed incomplete and patchy losses of S cones that were limited to the diabetic retinas. Statistically significant reduction in the density of S-cones was found at nearly all foveal eccentricities from 0.1 mm to 15.0 mm. This was not the case for the L/M-cones. On average, for all locations, the percentage of S-cones compared with L/M-cones was decreased by 21.0% +/- 3.4% with respect to the controls. CONCLUSION: The S-cones selectively die in diabetic retinopathy. CLINICAL RELEVANCE: Selective loss of S-cones may contribute to the tritan-like color vision deficit seen in patients with diabetic retinopathy. PMID- 11030823 TI - Argatroban inhibits intraocular fibrin formation after vitrectomy in rabbits. AB - OBJECTIVES: To examine whether the specific thrombin inhibitor argatroban can prevent anterior chamber, pupillary area, and anterior vitreous fibrin formation after vitrectomy and lensectomy in rabbits. METHODS: Argatroban was infused into the vitreous cavities of Japanese albino rabbits for 5 minutes after pars plana vitrectomy and lensectomy. Slitlamp microscopy and indirect ophthalmoscopy were performed at postoperative hours 0.5, 1, 2, 3, and 6, and at postoperative days 1, 2, 3, and 7, and the amounts of fibrin formation in the anterior chamber, pupillary area, and anterior vitreous were scored from grade 0 to 4. RESULTS: Argatroban prevented fibrin formation from 0.5 hours postoperatively in a dose dependent manner. In the eyes treated with 0.01% argatroban, the median score for postoperative fibrin formation was significantly less than that in control eyes between hours 1 and 3 (hour 1, P =.02; hour 2, P =.005; and hour 3, P =.003); the eyes treated with 0.003% argatroban also had significantly less fibrin than control eyes between 1 and 2 hours (hour 1, P =.005; hour 2, P =.03). CONCLUSION: These results indicate that argatroban inhibits intraocular fibrin formation in an experimental rabbit model. CLINICAL RELEVANCE: Argatroban may be useful clinically in cases that often produce fibrin postoperatively, such as proliferative vitreoretinopathy and proliferative diabetic retinopathy. PMID- 11030824 TI - Transformation of cell type in uveal melanomas: a quantitative histologic analysis. AB - OBJECTIVE: To describe the cytologic transformation and tumor progression in a series of uveal melanomas. METHODS: Fifteen cases of uveal melanoma, treated by primary transscleral local resection without primary adjuvant treatment, needed enucleation because of local tumor recurrence. Cytologic and cell morphometric features of the primary tumor and the intraocular recurrence were compared, with evaluation of the amounts of intermediate cells, epithelioid cells, mitotic figures, and nucleolar area. RESULTS: The cases were categorized into 2 groups, according to their cytologic characteristics. In the first group (5 cases), there was no cytopathological transformation in the recurrent tumor. The nucleolar area was increased in only 1 case. In the second group (10 cases), the recurrent tumors showed transformation into a more epithelioid cell type. In all but 1 case there was an increase in epithelioid cells in the tumor recurrence. The nucleolar area was increased significantly in all cases. The mean local recurrence interval in all cases was 15.3 months, with no difference between the groups. Death from metastases occurred in 7 cases in which the nucleolar area was 4.2 microm(2) in the primary tumor. CONCLUSIONS: These findings demonstrate that, in an individual tumor, the cytologic phenotype can change considerably even after a relatively short time, resulting in an increase in tumor-related mortality. CLINICAL RELEVANCE: Studies on the natural course of uveal melanoma have been very limited and based purely on observations on the progression of melanomas in terms of size and alteration of various clinical characteristics. PMID- 11030826 TI - New soft-tipped instruments for foveal translocation surgery with 360 degrees retinotomy. AB - Foveal translocation surgery with 360 degrees retinotomy involves several surgical procedures, including creation of a total retinal detachment, rotation of the retina, and complete reattachment of the retina. Some of these procedures, which are time-consuming and difficult to perform, may damage the retina when conventional surgical instruments are used. To make these procedures safer and easier, we have developed 3 soft instruments: an atraumatic forceps, a modified Chang double-barrel cannula with a silicone tip, and a light pipe with a silicone tip. The atraumatic forceps allows the surgeon to grasp and peel off the retina with less damage during the creation of retinal detachment. A bimanual technique using the other 2 instruments allows surgeons to rotate and reattach the detached retina easily and safely. We believe that these soft-tipped instruments make foveal translocation a less complicated surgery. PMID- 11030825 TI - Macular degeneration associated with aberrant expansion of trinucleotide repeat of the SCA7 gene in 2 Japanese families. AB - OBJECTIVE: To evaluate the macular function of Japanese patients with a trinucleotide repeat expansion in the spinocerebellar ataxia type 7 (SCA7) gene. METHODS: Ophthalmic findings in patients whose DNA analysis revealed expanded alleles of the trinucleotide repeat in the SCA7 gene were evaluated. RESULTS: Trinucleotide repeat was expanded from 40 to 48 in affected patients (control subjects, 12 repeats). Affected patients were characterized by different degrees of visual acuity decrease (0.09-0.9), a tritan axis color vision, a coarse granular appearance of the macular region on scanning laser ophthalmoscopy, depression of multifocal electroretinograms, and macular degeneration. However, pigmentary changes were not observed in the retina. The trinucleotide repeat was longer and the onset of macular dysfunction was earlier in the younger generation. One patient in a family manifested decreased visual acuity 10 years preceding other neurologic signs. CONCLUSIONS AND CLINICAL RELEVANCE: Patients with SCA7 mutations showed macular dysfunction or degeneration with expansion of CAG repeat in the SCA7 gene. However, the lesions were less pigmented than those previously reported. Patients also showed ophthalmologic anticipation, which has not been reported for the ocular changes in other patients who have trinucleotide repeat expansion of the responsible genes. PMID- 11030827 TI - Baruch spinoza: heretic, lens grinder. PMID- 11030828 TI - Submacular surgery: a millennium update. PMID- 11030829 TI - Ophthalmology in India. PMID- 11030830 TI - Ophthalmology in Spain. PMID- 11030831 TI - Ophthalmology in Portugal. PMID- 11030832 TI - Oculocerebral non-Hodgkin's lymphoma with uveal involvement: development of an epibulbar tumor after vitrectomy. AB - Primary ocular lymphoma is the ocular manifestation of primary oculocerebral non Hodgkin's lymphoma. We describe a 79-year-old woman with a 7-year history of bilateral uveitis and subsequent central nervous system lymphomas. Repeated diagnostic vitrectomy during the following 5 years failed to demonstrate intraocular lymphoma cells. Within 9 months after the second vitrectomy, an epibulbar tumor developed in the limbal region of the left eye at the site of the sclerotomy. The eye, blind and painful due to secondary angle-closure glaucoma, was enucleated. Histopathologically, the globe showed a diffuse large B-cell non Hodgkin's lymphoma extending from the ciliary body outward through the sclerotomy. We conclude that, following vitrectomy, a primary ocular lymphoma may extend through the sclerotomy lesion and present as an epibulbar tumor. Uveal involvement may occur in oculocerebral non-Hodgkin's lymphoma. PMID- 11030833 TI - Ultrastructural analysis of extraocular muscle in chronic progressive external ophthalmoplegia. AB - Extraocular muscles are primarily involved in many mitochondrial diseases, but no reports exist regarding the morphological appearance of the muscles in cases of long-standing ocular myopathies. For this reason, muscle samples obtained from surgery in a sporadic case of chronic progressive external ophthalmoplegia (CPEO) were used for ultrastructural investigation and molecular analysis of mitochondrial DNA. Genetic testing revealed a heteroplasmic macrodeletion of about 5.0 kilobases in length, localized between the 9570- and 14619-base pair regions. Electron microscopy revealed focal areas of both disruption and abnormality of mitochondria in only some of the muscle fibers, producing "selective vacuolization." This ultrastructural pattern was highly selective and limited to some extraocular muscle fibers, sparing all the others. The "selective damage" observed in this case of CPEO resembles that case occurring in another mitochondrial disease, Leber hereditary optic neuropathy, where damage occurs only in the papillomacular bundle of the retina, sparing peripheral axons. It is possible that some anatomical and physiological factors play a leading role in both Leber hereditary optic neuropathy and ocular myopathies. The ultrastructural aspect herein observed needs to be further investigated to better understand whether a particular muscle fiber type is the target of mitochondrial impairment in CPEO. PMID- 11030834 TI - Anti-inflammatory and healing properties of nerve growth factor in immune corneal ulcers with stromal melting. PMID- 11030835 TI - Retinal and subhyaloid hemorrhage as a complication of laser iridectomy for primary angle-closure glaucoma. PMID- 11030836 TI - Peripheral corneal infiltrates following oral diclofenac administration. PMID- 11030837 TI - Case reports and small case series: conjunctival cryptococcosis in the acquired immunodeficiency syndrome. PMID- 11030838 TI - Bilateral giant macular hole. PMID- 11030839 TI - Bilateral lymphomatous optic neuropathy diagnosed on optic nerve biopsy. PMID- 11030840 TI - Photo essay: phthiriasis palpebrarum. PMID- 11030841 TI - Photo essay: normal electro-oculogram in a patient with vitelliruptive macular dystrophy and multiple vitelliform cysts. PMID- 11030842 TI - Roberts pseudothalidomide syndrome. PMID- 11030844 TI - Report does not show that there is No relationship between disc appearance and sensitivity to intraocular pressure PMID- 11030843 TI - Report does not show that there is no relationship between disc appearance and sensitivity to intraocular pressure. PMID- 11030845 TI - Management of mid peripheral infiltrates in contact lens wearers. PMID- 11030846 TI - News and comment: october 2000 PMID- 11030847 TI - Remembering the patient. PMID- 11030848 TI - Higher-dose intravenous magnesium therapy for children with moderate to severe acute asthma. AB - OBJECTIVE: To evaluate the efficacy of a 40-mg/kg dose of intravenous magnesium sulfate for moderate to severe asthma exacerbations in pediatric patients. STUDY DESIGN: Double-blind placebo-controlled trial. SETTING: Two urban tertiary care pediatric emergency departments. SUBJECTS: Thirty patients, aged 6 to 17.9 years, being treated for an acute asthma exacerbation. INTERVENTION: Eligible patients received either a magnesium sulfate infusion of 40 mg/kg or saline solution. RESULTS: At 20 minutes, the time at which the infusion was completed, the magnesium group had a significantly greater percentage of absolute improvement from baseline in each of the following: predicted peak expiratory flow rate (8.6% vs 0.3%, P<. 001), forced expiratory volume in 1 second (7.0% vs 0.2%,P<.001), and forced vital capacity (7.3% vs -0.7%, P<.001). The improvement was greater at 110 minutes: peak expiratory flow rate (25.8% vs 1.9%, P<.001), forced expiratory volume in 1 second (24.1% vs 2.3%; P<. 001), and forced vital capacity (27.3% vs 2.6%, P<.001). Patients who received intravenous magnesium were more likely to be discharged to their homes than those who received the placebo (8/16 vs 0/14; P=. 002). CONCLUSION: Children treated with 40 mg/kg of intravenous magnesium sulfate for moderate to severe asthma showed remarkable improvement in short-term pulmonary function. PMID- 11030849 TI - Nebulizer use in inner-city children with asthma: morbidity, medication use, and asthma management practices. AB - OBJECTIVE: To assess the frequency of nebulizer use, describe morbidity and patterns of medication administration, and examine the potential relationships between inhaled anti-inflammatory medication administration, asthma morbidity, and asthma management practices in children with asthma using a nebulizer compared with children with asthma not using a nebulizer. RESEARCH DESIGN: A cross-sectional, descriptive survey of previous events. SETTING: Elementary schools and participants' homes in Baltimore, Md, and Washington, DC. PARTICIPANTS: Six hundred eighty-six families of children aged 5 to 12 years with a diagnosis of at least mild, persistent asthma. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Asthma morbidity, health care utilization, pattern of asthma medication administration, nebulizer use, and asthma management data were collected by telephone survey administered to caregivers. Nebulizer use was defined as use at least 1 or more days per month during the last 6 months. Of 686 children identified, 231 (33%) reported current nebulizer use. Nebulizer users had significantly increased lifetime hospital admissions, hospitalizations, and emergency department visits in the last 6 months compared with nonnebulizer users. Inhaled corticosteroid administration was low for both groups (nonnebulizer users, 8%; nebulizer users, 15%). In the nebulizer users group, administration of inhaled anti-inflammatory medications was associated with increased asthma morbidity (increased hospitalizations, days and nights with symptoms, and oral steroid use). CONCLUSIONS: Nebulizer use by inner-city children with asthma is higher than anticipated but is not associated with reduced asthma morbidity. This group of high-risk children was undertreated with inhaled corticosteroids for long-term control of asthma despite reports of adequate monitoring by a primary care physician. PMID- 11030850 TI - Asthma and bronchiolitis hospitalizations among American Indian children. AB - OBJECTIVE: To compare asthma and bronchiolitis hospitalization rates in American Indian and Alaskan native (AI/AN) children and all children in Washington State. METHODS: A retrospective data analysis using Washington State hospitalization data for 1987 through 1996. Patients were included if asthma or bronchiolitis was the first-listed diagnosis. American Indian and Alaskan native children were identified by linking state hospitalization data with Indian Health Service enrollment data. RESULTS: Similar rates of asthma hospitalization were found for AI/AN children older than 1 year compared with all children. In AI/AN children younger than 1 year, hospitalization rates for asthma (528 per 100,000 population; 95% confidence interval [CI], 346-761) and bronchiolitis (2954 per 100,000 population; 95% CI, 2501-3456) were 2 to 3 times higher than the rates in all children (232 per 100,000 population [95% CI, 215-251] and 1190 per 100,000 population [95% CI, 1149-1232], respectively). Hospitalization rates for asthma and bronchiolitis increased 50% between 1987 and 1996 for all children younger than 1 year and almost doubled for AI/AN children younger than 1 year. CONCLUSIONS: American Indian and Alaskan native children have significantly higher rates of hospitalization for wheezing illnesses during the first year of life compared with children of other age groups and races. Furthermore, the disparities in rates have increased significantly over time. Future public health measures directed at managing asthma and bronchiolitis should target AI/AN infants. PMID- 11030851 TI - Duration of illness in ambulatory children diagnosed with bronchiolitis. AB - OBJECTIVES: To measure the duration of illness in ambulatory children diagnosed with bronchiolitis and to examine clinical predictors of duration of illness. DESIGN: Validation inception cohort study. Duration of follow up was 28 days. SETTING: A primary-level ambulatory department of a public sector children's hospital in Cape Town, South Africa. PATIENTS: One hundred eighty-one children aged 2 to 23 months who went to the hospital as their first contact for that episode of illness, and had a clinical diagnosis of bronchiolitis were enrolled consecutively on weekday mornings if their guardian stated that they were contactable by telephone. MAIN OUTCOME MEASURE: Resolution of symptoms, as judged by the guardian, measured by twice-weekly telephone interviews. RESULTS: Median duration of illness (calculated as the reported duration of symptoms before initial hospital visit plus the time from first consultation to recovery) was 12 days (95% confidence interval, 11-14 days). After 21 days, 18% were still ill and after 28 days, 9% were still ill. Sixty-two patients (34.2%) had unscheduled consultations within 28 days, a median of 13 days after the first consultation. There was no association of duration of illness with age, sex, z score for weight for age, or respiratory rate. CONCLUSIONS: Ambulatory children diagnosed with bronchiolitis recover with few complications, but the resolution of symptoms may take several weeks. Providing parents with this information could help reduce the high rate of unscheduled return visits as observed in this cohort. PMID- 11030852 TI - Sustaining the implementation of an evidence-based guideline for bronchiolitis. AB - OBJECTIVE: To describe the changes occurring over a 3-year period after implementation of an evidence-based clinical practice guideline for the care of infants with bronchiolitis. DESIGN: Before and after study. SETTING: Children's Hospital Medical Center, Cincinnati, Ohio. PATIENTS: Infants 1 year or younger admitted to the hospital with a first-time episode of typical bronchiolitis. INTERVENTION: The guideline was implemented January 15, 1997. Data on all patients discharged from the hospital with bronchiolitis, from January 15 through March 27, in 1997, 1998, and 1999, were stratified by year and compared with data on similar patients discharged from the hospital in the same periods in the years 1993 through 1996. MAIN OUTCOME MEASURES: Patient volumes, length of stay for admissions, and use of specific laboratory and therapeutic resources ancillary to bed occupancy. RESULTS: After implementation of the guideline, admissions decreased 30% and mean length of stay decreased 17% (P<.001). Nasopharyngeal washings for respiratory syncytial virus were obtained in 52% fewer patients (P<.001); 14% fewer chest x-ray films were ordered (P<.001). There were significant reductions in the use of all respiratory therapies, with a 17% decrease in the use of at least 1 beta(2)-agonist inhalation therapy (P<.001). In addition, 28% fewer repeated inhalations were administered (P<.001); mean costs for all resources ancillary to bed occupancy fell 41% (P<.001); and mean costs for respiratory care services fell 72% (P<.001). CONCLUSIONS: An evidence-based clinical practice guideline for the care of patients encountered in major pediatric care facility has been successfully sustained beyond the initial year of its introduction to practitioners in southwest Ohio. PMID- 11030853 TI - The recognition of facial expression of pain in full-term newborns by parents and health professionals. AB - OBJECTIVE: To determine whether adults can recognize neonatal facial expression of pain. DESIGN: A cross-sectional study. SETTING: Neonatal intensive care unit, nursery, and outpatient clinic of one university hospital and one private hospital in Sao Paulo, Brazil. PATIENTS: Four hundred five adults divided into 2 groups: health and nonhealth professionals. INTERVENTION: The faces of 3 healthy full-term newborns who needed glucose screening were photographed at rest and during light exposure, heel rubbing, and heel puncture. A series of adults answered a questionnaire on personal and professional data and then they analyzed for 1 minute each of the 3 sets of pictures to answer the following question: "In which picture of this set do you think that the baby is feeling pain?" MAIN OUTCOME MEASURE: Number of correct answers for the 3 sets of photographs shown to the adults. RESULTS: Seventy-four percent of the health professionals and 86% of the nonhealth professionals indicated correctly the picture with facial expressions of pain in at least 2 of the 3 sets. Regarding which picture was picked out by the interviewee, 94% of the health professionals and 92% of the nonhealth professionals indicated the picture taken during the heel puncture in set 1. The same observation was made by 53% and 54% of the health professional and by 68% and 66% of the nonhealth professional interviewees for sets 2 and 3, respectively. CONCLUSIONS: Facial expression of pain represents an effective neonatal communication tool. However, the health professional group achieved a lower level of recognition of neonatal facial expressions of pain. Factors related to the personal and professional characteristics of the adults interviewed probably contributed to this result. PMID- 11030854 TI - Economic evaluation of Safer Choices: a school-based human immunodeficiency virus, other sexually transmitted diseases, and pregnancy prevention program. AB - OBJECTIVE: To evaluate the cost-effectiveness and cost benefit of Safer Choices, a school-based human immunodeficiency virus, other sexually transmitted diseases, and unintended pregnancy prevention intervention for high school students. METHODS: The baseline cost-effectiveness and cost benefit were derived in 4 steps: (1) estimation of intervention costs; (2) adaptation of the Bernoulli model to translate increases in condom use into cases of human immunodeficiency virus and other sexually transmitted diseases averted, and development of a model to translate increases in contraceptive use into cases of pregnancy averted; (3) translation of cases averted into medical costs and social costs averted; and (4) calculation of the net benefit of the program. Multivariable sensitivity analysis was performed to determine the robustness of the base-case results. RESULTS: Under base-case assumptions, at an intervention cost of $105,243, Safer Choices achieved a 15% increase in condom use and an 11% increase in contraceptive use within 1 year among 345 sexually active students. An estimated 0.12 cases of human immunodeficiency virus, 24.37 cases of chlamydia, 2.77 cases of gonorrhea, 5.86 cases of pelvic inflammatory disease, and 18.5 pregnancies were prevented. For every dollar invested in the program, $2.65 in total medical and social costs were saved. Results of most of the scenarios remained cost saving under a wide range of model variable estimates. CONCLUSIONS: The Safer Choices program is cost effective and cost saving in most scenarios considered. School-based prevention programs of this type warrant careful consideration by policy makers and program planners. Program cost data should be routinely collected in evaluations of adolescent prevention programs. PMID- 11030855 TI - Health-compromising behaviors: why do adolescents smoke or drink?: identifying underlying risk and protective factors. AB - OBJECTIVES: To better understand the motivation for adolescent smoking and drinking and to identify the underlying risk and protective factors associated with these behaviors among adolescents. DESIGN: Cross-sectional, school-based survey of students in grades 5 through 12. PARTICIPANTS: A nationally representative sample of 2574 boys and 2939 girls in grades 7 through 12 from 297 public, private, and parochial schools across the United States who participated in The Commonwealth Fund Survey of the Health of Adolescent Girls and Boys in 1997. MAIN OUTCOME MEASURES: Sex-specific adjusted relative risks (RRs) and 95% confidence intervals (CIs) comparing self-reported regular smokers and regular drinkers by risk and protective factors with adolescents reporting none of these behaviors. RESULTS: Adolescent boys and girls were equally likely to be regular smokers (11.2%). The prevalence rate of regular drinking was only slightly higher for boys (22.4%) than it was for girls (19.3%). The rates of both health-risk behaviors were significantly higher for those reporting risk factors, and the strengths of associations varied by sex. Sex differences also emerged in motivation for engaging in these behaviors. When we adjusted for demographic characteristics, exposure to childhood abuse (RR, 4.1; 95% CI, 2.4-7.0) and stressful life events (RR, 2.4; 95% CI, 1.1-5.4) were strongly associated with increased risk for boys' regular smoking. Similar associations were found for regular drinking. For girls, a history of abuse (RR, 1.8; 95% CI, 1.1-2.8), violence within the family (RR, 2.2; 95% CI, 1.6-3.2), depressive symptoms (RR, 1.6; 95% CI, 1.0-2.4), and stressful life events (RR, 3.1; 95% CI, 1.8-5.6) were significantly associated with increased risk for regular smoking. Similar associations were again found for regular drinking. Parental support was protective against both health-risk behaviors for both sexes. Participation in extracurricular activities was associated with lower risk for regular smoking for boys (RR, 0.4; 95% CI, 0.2-0.7) and for girls (RR, 0.3; 95% CI, 0.2-0.5); however, there was no significant association between drinking behavior and participation in activities. CONCLUSIONS: The increased risk for regular smoking and regular drinking among adolescents with a history of abuse, family violence, depressive symptoms, and stressful life events suggests that routine screening for abuse, violence, and other family experiences should be an essential component of adolescent health care visits. Effective prevention programs to reduce smoking and drinking among adolescents should recognize that health-risk behaviors may be associated with other negative life experiences and that the strength of these associations differs by sex. PMID- 11030856 TI - Adolescent violence prevention practices among California pediatricians. AB - OBJECTIVES: To examine pediatricians' provision of violence prevention services to their adolescent patients and to identify factors associated with pediatricians' implementation of these services. DESIGN: A cross-sectional sample of California pediatricians completed a self-report questionnaire. The "Precede/Proceed" theoretical model guided the questionnaire in identification of factors associated with pediatricians' screening and intervening practices in preventing adolescent violence. RESULTS: Two hundred twenty pediatricians (54% female, 66% white, 24% Asian, 5% Latino, and 5% other) participated in the study. On average, participants screened their patients for violence-related risk factors 31% of the time for fighting, 39% of the time for violence in the home, and 29% of the time for weapon carrying. Participants provided their at-risk patients with violence-related interventions less than 50% of the time (on average) implementing the following interventions: written materials, follow-up appointments, discipline counseling, or referral to a community organization, Child Protective Services, or a specialized adolescent clinic. Factors associated with violence prevention screening practices included the following: positive attitudes and beliefs regarding screening for violence, familiarity with violence prevention guidelines, use of prompts in medical records, perceptions of greater skills, and positive reinforcement from patients and colleagues for providing violence prevention services (R(2)=0.44; P<.001). Factors associated with violence prevention intervention practices included: positive attitudes and beliefs in screening for violence, availability of resources, and positive reinforcement from patients and colleagues for providing violence prevention services (R(2)=0.37; P<.001). CONCLUSIONS: California pediatricians are not widely screening their adolescent patients for risk factors associated with adolescent violence, nor are they providing interventions to their adolescent patients who may be at risk for violence. The factors associated with pediatricians' implementation of violence prevention services may assist in the development of effective interventions designed to enhance their delivery of these services to their adolescent patients. PMID- 11030857 TI - Functional limitations and key indicators of well-being in children with disability. AB - OBJECTIVES: To compare measures of well-being in children with and without different types and severity of limitations. DESIGN: Nationally representative data for American children aged 5 to 17 years were drawn from the 1994 and 1995 National Health Interview Surveys on Disability (NHIS-D) (N = 41,300) and the Year 2000 Health Supplement to the 1994 NHIS-D (N = 9530). Family resources, safety, health status, and health access were measures of environment. The presence and severity of limitations were measured in the domains of mobility, self-care, communication, and learning. RESULTS: Children with functional limitations were more likely to live in families with limited resources and have greater exposure to secondhand smoke, less access to health care, and lower health status. Children with a limitation were not less likely to have a regular source of medical care, but they more often were delayed or prevented from getting necessary health care due to cost or insurance. CONCLUSIONS: Standard measures of child well-being were appropriate for children with functional limitations and showed their unfavorable situations. Children with functional limitations more often have unfavorable family resources, less healthy home environments, poorer health status, and less health service access than other children, making them more susceptible to developmental difficulties beyond those difficulties associated with the challenges of their specific functional limitations. PMID- 11030858 TI - Teaching resuscitation to pediatric residents: the effects of an intervention. AB - OBJECTIVE: To evaluate the effectiveness of an educational intervention on pediatric residents' resuscitation fund of knowledge, technical skills, confidence, and overall performance. DESIGN: Prospective, nonconcurrent, controlled interventional trial. SETTING: Urban pediatric tertiary care hospital. PARTICIPANTS: An intervention group (IG) of 28 pediatric residents graduating in 1997, and a control group (CG) of 30 pediatric residents graduating in 1996. INTERVENTIONS: Resuscitation course with didactic lectures and skills practice stations, as well as a minimum of 3 practice mock resuscitations with immediate feedback throughout postgraduate year 3. MAIN OUTCOME MEASURES: Fund of knowledge, using the Pediatric Advanced Life Support test and short answer test; technical skills, using the Airway and Vascular Access Skills Assessment; experience and confidence, using an anonymous survey; and overall performance, evaluated using a videotaped mock resuscitation test. RESULTS: The IG scored better on the short answer test (P<.001). A larger number of IG residents were successful in the completion of ancillary airway maneuvers and femoral vascular access (P =.02), as well as endotracheal intubation (P =.004) and intraosseous access (P =.002). The IG was more confident in their leadership role (P =.0001) and technical skills (P =.05). Trends toward improved overall performance were noted for the IG mock resuscitations. Residents in the IG were more likely to assess the airway in fewer than 2 minutes (P =.02), recognize the threat to life in fewer than 5 minutes (P =.02), and complete the primary survey in a timely fashion (P =.05). They required fewer prompts (P =.04) and made fewer mistakes (P =.07). CONCLUSIONS: A structured, formal curriculum can improve the necessary fund of knowledge, skills, confidence, and leadership required for resuscitation. PMID- 11030859 TI - Picture of the month. Sarcoidosis with bone involvement. PMID- 11030860 TI - Pathological case of the month. Eosinophilic granuloma (Langerhans cell histiocytosis). PMID- 11030861 TI - Radiological case of the month. Multiple-bone tuberculosis and dactylitis. PMID- 11030862 TI - The pediatric forum: spontaneous passage of esophageal coins questionable PMID- 11030863 TI - Spontaneous passage of esophageal coins questionable. PMID- 11030864 TI - Chiropractic care for children can help. PMID- 11030865 TI - The pediatric forum: chiropractic care for children can help PMID- 11030866 TI - In defense of pediatric chiropractic care. PMID- 11030867 TI - Universal childhood vaccinations: a Faustian bargain? PMID- 11030868 TI - Complications of laparoscopic splenectomy. AB - HYPOTHESIS: Analysis of the type and characteristics of complications after laparoscopic splenectomy may permit the identification of clinical factors with predictive value for the development of complications. DESIGN: Univariate and multivariate analysis of factors related to complications in a prospective series of laparoscopic splenectomies. SETTING: A large tertiary referral university teaching general hospital. PATIENTS: One hundred twenty-two nonselected consecutive patients, in whom laparoscopic splenectomy was attempted between February 1993 and July 1999. INTERVENTION: Laparoscopic splenectomy. MAIN OUTCOME MEASURES: Immediate complications classified according to the Clavien score. Univariate and multivariate analyses were performed of complications related to age, sex, body mass index, and malignant nature of the hematologic disease; preoperative hematocrit and platelet count; operative time; operative position; need of accessory incision; transfusion status; learning curve; and existence of comorbid diseases. RESULTS: One hundred thirteen laparoscopic splenectomies were completed (conversion rate, 7.4%). Twenty patients (18%) developed 23 complications. All were Clavien type I or II, without mortality. One complication was intraoperative (diaphragmatic perforation), and 22 were postoperative: 6 pulmonary (26%), 3 fever (13%), 8 hemorrhagic (35%) (5 episodes of postoperative bleeding and 3 abdominal wall hematomas), and 6 others (26%). Ten (43%) of the 23 were technically related. Univariate analysis showed that complications were only related to age (mean +/- SD, 55 +/- 15 vs 39 +/- 17 years; P<.008) or transfusion (50% vs 11%; P<.001). Multivariate analysis showed that the learning curve (P<.005; 95% confidence interval, 2.46), age (P<.001; 95% confidence interval, 1. 04), spleen weight (P<.009; 95% confidence interval, 1.00), and malignant neoplasm diagnosis (P<.007; 95% confidence interval, 3.82) were independent predictors of complications. CONCLUSIONS: Laparoscopic splenectomy is feasible, and the incidence of severe complications is reduced. However, a high proportion of these complications are technique related. Laparoscopic splenectomy requires great technical care but offers major clinical advantages, even in less favorable situations, such as in patients with splenomegaly or with malignant neoplasms. PMID- 11030869 TI - Apoptosis and surgical trauma: dysregulated expression of death and survival factors on peripheral lymphocytes. AB - BACKGROUND: Surgery and anesthesia cause depression of cell-mediated immunity in the postoperative period, including a reduction in the numbers of circulating lymphocytes. It has been claimed that this immunosuppression is associated with an increased incidence of postoperative infections. HYPOTHESIS: Lymphocytopenia following surgical trauma depends on a dysregulated expression of death/and survival factors associated with apoptosis that, in turn, interferes with the occurrence of postsurgical infections. DESIGN: Fifteen subjects undergoing elective surgery under general anesthesia entered the study. The data of the patients who had infections during the postoperative outcome were compared with the data of those who did not. The data were collected prospectively. MAIN OUTCOME MEASURES: Peripheral blood samples were drawn before the operation, and 24 hours and 96 hours after the operation. Lymphocytes were isolated and examined for quantification and phenotypic analysis of apoptosis using the 7-amino actinomycin D method, as well as for Fas and Fas ligand, interleukin 1-converting enzyme p20/caspase-1, Bcl-2, and p35 expression. The rate of apoptotic cells was correlated with the incidence of postoperative infections. SETTING: University hospital. RESULTS: Twenty-four hours after surgery, CD4(+) and CD8(+) cells exhibited a significantly higher frequency of apoptosis as well as of Fas and Fas ligand and interleukin 1-converting enzyme p20/caspase-1 expressions than preoperatively. This increase was paralleled by a significant down-regulation of antiapoptotic factors such as Bcl-2. However, the expression of the proapoptotic factor p35 was reduced. In addition, we found a relationship between the rate of the apoptotic CD8(+) subset and the occurrence of infectious complications during the postoperative course. At 96 hours after surgery, the variables studied returned to the baseline levels. CONCLUSIONS: In the early postoperative period, surgical trauma under general anesthesia induces an intracellular perturbation on peripheral lymphocytes, resulting in both up-regulation of death-signaling factors and down-regulation of survival-signaling factors. The increased apoptosis of CD8(+) lymphocytes, but not of CD4(+) cells, seemed to be associated with a greater risk of postsurgical infections. PMID- 11030870 TI - Enhancement of wound healing by hyperbaric oxygen and transforming growth factor beta3 in a new chronic wound model in aged rabbits. AB - HYPOTHESIS: Although hyperbaric oxygen (HBO) has been used clinically for 3 decades, there have been few controlled clinical trials. Animal models have not been adequate to test the efficacy of HBO in the treatment of chronic wounds, either by itself or in combination with growth factors. We hypothesize that HBO is as efficacious as a prototype growth factor in improving wound healing in a new animal model of ischemic chronic wounds. DESIGN: Twenty-five aged rabbits and 3 young rabbits had their ears rendered chronically ischemic and ulcers were created down to the level of cartilage. These ulcers were treated in 1 of 3 ways: with HBO, 90 minutes per day, Monday through Friday, for 4 weeks; with transforming growth factor beta(3) at 1 microg/cm(2); or with both modalities combined. Controls were treated with vehicle or hyperbaric room air or both. RESULTS: This model created an aged/ischemic wound that failed to heal spontaneously up to 26 days after wounding (88% reduction compared with aged/nonischemic controls). Hyperbaric oxygen alone and transforming growth factor beta(3) alone both improved healing rate (only 38% reduction in healing compared with aged/nonischemic controls). Combined therapy produced no additional improvement over either modality by itself. CONCLUSIONS: In aged animals, HBO and transforming growth factor beta(3) were equally effective in improving wound healing. Our data suggest that HBO alone may be more effective in the chronic wound than in the acute wound. There was no additive benefit to combining modalities as has been reported in the same wound model in young rabbits. PMID- 11030871 TI - Transforming growth factor beta3 promotes fascial wound healing in a new animal model. AB - HYPOTHESIS: Transforming growth factor beta(3) (TGF-beta(3)) promotes fascial wound healing in a new animal model, as measured by wound breaking strength, collagen deposition, and cellular proliferation. DESIGN/INTERVENTION: Bilateral, longitudinal incisions were made in the anterior rectus sheaths of 24 male New Zealand white rabbits. One incision was treated with 1 microg of TGF-beta(3); the contralateral incision served as a control. The wounds were harvested at 1, 2, 3, 4, 6, and 8 weeks after creation ("wounding"). MAIN OUTCOME MEASURES: Wound tissue was tested for breaking strength using a tensiometer and processed for histological examination of collagen deposition and cellular proliferation at all time points after wounding. Collagen deposition and cellular proliferation were measured in histological cross sections of wounds with Masson trichrome staining and proliferating cell nuclear antigen immunohistochemistry, respectively. RESULTS: At all time points after wounding, treatment with TGF-beta(3) significantly increased the wound breaking strength (up to 138%) and collagen deposition (up to 150%) over the control group. Cellular proliferation was increased during the first 3 weeks after wounding (up to 147%), but returned to baseline levels by the fourth week. CONCLUSIONS: Transforming growth factor beta(3) promotes fascial wound healing. In this new animal model of fascial wound healing, TGF-beta(3) increased fascia breaking strength, collagen deposition, and cellular proliferation. These results are similar to findings in cutaneous wound models and demonstrate, for the first time, a pharmacologic agent to accelerate fascial healing. PMID- 11030872 TI - Electrical stimulation-induced changes in double-wrapped muscles for dynamic graciloplasty. AB - HYPOTHESIS: Treatment of fecal incontinence has been greatly improved by electrical stimulation of gracilis muscle transposed around the anal canal. Various configurations of the muscle have been used: single alpha, gamma, epsilon muscle loops, split sling, or double wrap. We report herein experimental data on muscle transformation and damage induced by the latter surgical approach. DESIGN, INTERVENTIONS, AND MAIN OUTCOME MEASURES: This study was conducted on 4 groups of New Zealand white rabbits. Group 1 had unstimulated transposed gracilis muscles. Group 2 had left transposed gracilis muscles stimulated only. Group 3 had both right and left transposed gracilis muscles stimulated. Group 4 were the controls (not operated on). Muscle properties were studied by electrophysiological,immunohistochemical,and biochemical techniques. RESULTS: Transformation from fast-contractile glycolytic muscle fibers into fast intermediate to slow-contractile oxidative muscle fiber types induced a fatigue resistance of the transposed muscle that has undergone long-term stimulation and muscle alterations characterized by fiber atrophy and fibrosis. CONCLUSIONS: Whatever technique of dynamic graciloplasty is used, muscle degeneration associated with mobilization might result primarily from the surgical dissection, whereby collateral blood supply to the gracilis is interrupted and exacerbated by long-term stimulation. PMID- 11030873 TI - Interval nodes: the forgotten sentinel nodes in patients with melanoma. AB - BACKGROUND: Any sentinel lymph node that receives lymph drainage directly from a primary melanoma site, regardless of its location, may contain metastatic disease. This is true even if the sentinel node does not lie in a recognized node field. Interval (in-transit) nodes that lie along the course of a lymphatic vessel between a primary melanoma site and a recognized node field are sometimes seen during lymphatic mapping for sentinel node biopsy. If drainage to such interval nodes is ignored by the surgeon during sentinel node biopsy, metastatic melanoma will be missed in some patients. HYPOTHESIS: When lymph drains directly from a cutaneous melanoma site to an interval node, that sentinel node has the same chance of harboring micrometastatic disease as a sentinel node in a recognized node field. DESIGN: Preoperative lymphoscintigraphy with technetiumTc 99m antimony trisulfide colloid was performed to define lymphatic drainage patterns and, since 1992, to locate the sentinel lymph nodes for surgical biopsy or for permanent skin marking of their location with point tattoos. SETTING: Melanoma unit of a university teaching hospital. PATIENTS: A total of 2045 patients with cutaneous melanoma were studied in 13 years. RESULTS: Interval nodes were found in 148 patients (7.2%). The incidence of interval nodes varied with the site of the primary melanoma. Interval nodes were more common with melanomas on the trunk than with those on the lower limbs. Micrometastatic disease was found in 14% of interval nodes that underwent biopsy as sentinel nodes. This incidence is similar to that found in sentinel nodes located in recognized node fields, confirming the potential clinical importance of interval nodes. CONCLUSIONS: Interval nodes should be removed surgically along with any additional sentinel nodes in standard node fields if the sentinel node biopsy procedure is to be complete. In some patients, an interval node will be the only lymph node that contains metastatic disease. PMID- 11030874 TI - Pediatric restraint use in motor vehicle collisions: reduction of deaths without contribution to injury. AB - HYPOTHESIS: Restraint use for children in automobiles is mandated in every state, but injury patterns are unknown. Although use of pediatric retraints is associated with reducing morbidity and mortality, the injury distribution for specific anatomic sites may be altered in restrained vs unrestrained children. DESIGN: Review of trauma registry data, medical records, and autopsy findings. SETTING: Urban level I trauma center and tertiary care children's hospital. PATIENTS: All children aged 6 years or younger who were in motor vehicle collisions from June 1, 1990, through March 31, 1997. MAIN OUTCOME MEASURES: Age, weight, restraint use and type, collision data, Injury Severity Score (ISS), injury type, and outcome. RESULTS: We included 600 children. The restrained group showed a reduction in severe injuries for every anatomic site and had a lower mean ISS, fewer injuries, and more uninjured children. The restrained group also had a reduction in the incidence of hollow- and solid-organ abdominal injuries. CONCLUSIONS: Age-appropriate restraint devices decrease mortality and reduce the incidence of significant injury in motor vehicle collisions for all anatomic sites in young children. In contrast to injuries attributed to restraint use in adults, specific restraint-related injury patterns were not seen in children. PMID- 11030877 TI - Invited critique PMID- 11030875 TI - Increased expression of intestinal P-selectin and pulmonary E-selectin during intravenous total parenteral nutrition. AB - HYPOTHESIS: Intravenous total parenteral nutrition (TPN) induces intestinal polymorphonuclear neutrophil recruitment with increased intestinal intercellular adhesion molecule-1 expression. While intercellular adhesion molecule-1 causes firm adhesion of leukocytes to the endothelial cells, P- and E-selectin mediate leukocyte recruitment via rolling. Therefore, manipulation of nutrition may also affect P- and E-selectin expression in organs. DESIGN: Prospective randomized experimental trials. SETTING: Laboratory. MATERIALS: Male mice. INTERVENTIONS: Fifty-three mice were randomized to chow, intravenous TPN, or intragastric TPN. MAIN OUTCOME MEASURES: After 5 days of diet, mice were administered iodine 125 labeled anti-P-selectin antibody (or iodine 125-labeled anti-E-selectin antibody) and iodine 131-labeled nonbinding antibody to quantify P-selectin (or E-selectin) expression in organs (lung, liver, kidney, small intestine, colon, stomach, pancreas, mesentery, heart, and skeletal muscle). RESULTS: P-selectin in small intestine, colon, stomach, and pancreas in the intravenous TPN group increased significantly as compared with the chow and the intragastric TPN groups. E selectin expression was up-regulated after intravenous TPN in the lung but not in other sites. CONCLUSIONS: In a time frame (5 days) when intercellular adhesion molecule-1 expression and neutrophil recruitment are increased, intestinal expression of P-selectin remains up-regulated. Early lung inflammatory changes are reflected by increases in E-selectin. This change may reflect early pulmonary dysfunction with intravenous TPN, but its significance requires further study. PMID- 11030876 TI - An improved model of acute liver failure based on transient ischemia of the liver. AB - HYPOTHESIS: A reproducible and potentially reversible model of acute liver failure in the pig is feasible based on transient ischemia of the liver. DESIGN: To determine the shortest period of liver ischemia sufficient to cause 100% mortality, ischemia of the liver was induced for different lengths of time, starting with 6 hours. If the pig survived, ischemia time was prolonged for 2 hours in the next animal. In the first group, the common bile duct was not tightened. In the second group, the common bile duct was tightened. SETTING: The Laboratory for Hepatopathophysiology, Catholic University, Leuven, Belgium. PARTICIPANTS: Female stress-negative Belgian Landrace pigs weighing 18 to 22 kg. INTERVENTIONS: During preparatory surgery, all ligaments around the liver and connective tissue around the liver hilum were transected and an end-to-side portacaval shunt was made. Vessel loops were placed around the branches of the hepatic artery and bile duct. Three days later, in fully awake pigs, the loops were tightened. MAIN OUTCOME MEASURES: Mortality. Development of acute liver failure was determined based on neurologic, biochemical, and pathological variables. RESULTS: When occluded for 10 hours, all pigs in group 2 (n = 5) [corrected] died between 12 and 17 hours after the induction of ischemia. All pigs developed typical acute liver failure. Tissue specimens showed 90% necrosis of the liver parenchyma. CONCLUSION: A highly reproducible and potentially reversible model of acute liver failure in the large animal has been established. PMID- 11030878 TI - Diabetic mastopathy: a report of 5 cases and a review of the literature. AB - BACKGROUND: Diabetic mastopathy is an unusual fibroinflammatory breast lesion that characteristically presents in premenopausal women with long-standing type 1 diabetes mellitus with multiple microvascular complications. The pathogenesis of this condition is believed to involve an autoimmune reaction to the accumulation of abnormal matrix induced by hyperglycemia. Clinicopathologic features include the development of dense keloidlike breast masses that are often recurrent or bilateral or both. Clinical distinction from a malignancy can be difficult. However, the benign nature of this lesion is easily recognized on histologic examination, and it is not associated with an increased incidence of epithelial or stromal neoplasia. HYPOTHESIS: A constellation of histopathologic and clinical features is necessary to make the diagnosis of diabetic mastopathy. Unnecessary surgery can be avoided in the clinical follow-up of patients with multiple, bilateral, and recurrent lesions. DESIGN: Case series. PATIENTS AND METHODS: Between December 1993 and December 1998, 5 premenopausal women with type 1 diabetes mellitus of 18 to 23 years' duration presented with nontender, palpable, firm-to-hard breast masses. To date, progression of the tumorlike proliferations has been bilateral and recurrent in 2 patients, bilateral in a third patient, and recurrent in a fourth. The fifth patient has developed neither bilateral nor recurrent lesions. Imaging studies did not in any patient demonstrate a focal lesion. All lesions were treated by either excisional (4 patients) or core (1 patient) biopsy. The resected specimens were examined histopathologically. RESULTS: Gross examination of the specimens showed firm masses with homogeneous tannish-white cut surfaces. They measured between 3.0 and 6.0 cm in maximum diameter. Microscopic examination showed keloidal fibrosis with ductitis, lobulitis, and vasculitis. The clinical profile in combination with these pathologic features is characteristic of diabetic mastopathy. CONCLUSIONS: Physicians should be aware of the association of long-standing diabetes mellitus with the development of benign fibroinflammatory breast lesions when managing these in premenopausal women. We outline the constellation of findings on clinical examination, medical history, imaging studies, and histopathologic examination that are required to make the diagnosis of diabetic mastopathy. Although these breast masses may be recurrent, they are not premalignant. In the appropriate setting, the diagnosis can be made by core biopsy, avoiding unnecessary surgeries in patients with multiple, bilateral, or recurrent lesions. PMID- 11030880 TI - Invited critique PMID- 11030879 TI - Indications for bilateral modified radical neck dissection in patients with papillary carcinoma of the thyroid. AB - HYPOTHESES: After subtotal thyroidectomy with modified radical neck dissection of the affected side, nodal recurrence at the contralateral cervical side indicates a poor prognosis for patients with papillary thyroid cancer. Bilateral modified radical neck dissection is beneficial for patients at high risk for contralateral nodal recurrence. DESIGN AND SETTING: Retrospective study of patients with papillary cancer who were treated surgically from January 1, 1970, through December 31, 1995, at the Noguchi Thyroid Clinic and Hospital Foundation, Beppu, Japan. PATIENTS: Patients (N = 1776) had primary tumors greater than 10 mm in maximum diameter and underwent thyroidectomy and ipsilateral modified radical neck dissection with curative intent. RESULTS: Thirty-two patients (1.8%) developed contralateral lymph node metastases during the mean follow-up period of 12.1 years. The risk factors for contralateral nodal recurrence were male sex, large primary tumor, tumor extension over the isthmus, extracapsular adhesion or invasion to surrounding tissues, and the presence of gross nodal metastasis at initial surgery. These patients had a greater number of distant metastases (31.1% vs 0.7%; P<.001) and a lower 10-year survival rate (83.7% vs 99.3%; P<.001) than patients without nodal recurrence. CONCLUSION: Bilateral modified radical neck dissection should be considered for patients with papillary carcinoma who show risk factors for contralateral nodal recurrence, as it could prevent a second operation and may improve their outcome. PMID- 11030882 TI - Invited critique PMID- 11030881 TI - Effective control of hepatic bleeding with a novel collagen-based composite combined with autologous plasma: results of a randomized controlled trial. AB - HYPOTHESIS: A novel collagen-based composite of bovine microfibrillar collagen and bovine thrombin combined with autologous plasma is more effective than standard hemostasis (collagen sponge applied with pressure) in controlling diffuse hepatic bleeding after hemihepatectomy or segmental resection of the liver. DESIGN: Randomized controlled trial. SETTING: Seven university-affiliated medical centers. PATIENTS: Sixty-seven adult patients scheduled for hemihepatectomy or segmental resection who received hemostatic intervention with an investigational treatment (n = 38) or control (n = 29). INTERVENTION: Bleeding hepatic tissue was managed in all control subjects with a collagen sponge with manual pressure. Subjects in the experimental group had the sprayable liquid composite intraoperatively applied to the surgical site. The liquid immediately formed a collagen-fibrin gel that was used without concomitant tamponade. MAIN OUTCOME MEASURES: Hemostatic success was defined as the proportion of subjects in each treatment group who achieved complete hemostasis within 10 minutes. Success rates and median times required to achieve controlled bleeding (ie, slight oozing) and complete hemostasis were compared between treatment groups. RESULTS: All 38 subjects in the experimental group achieved complete hemostasis within 10 minutes compared with only 69% (20/29) of control subjects (P<.001). The median time to controlled bleeding was approximately 4 times longer (250 vs 62 seconds) for control subjects than for experimental group subjects (P<.001). The median time required to achieve complete hemostasis also favored the experimental group (150 vs 360 seconds; P<.001). No adverse events related to the use of the experimental hemostatic agent were detected. CONCLUSIONS: The experimental composite is more effective at controlling and stopping diffuse hepatic bleeding than a collagen sponge applied with pressure; it may be a useful hemostatic agent for patients undergoing hemihepatectomy, segmental resection, and related surgical procedures. PMID- 11030883 TI - Clostridium difficile toxins influence hepatocyte protein synthesis through the interleukin 1 receptor. AB - HYPOTHESIS: Clostridium difficile toxins require interleukin 1 (IL-1) production or a functioning IL-1 receptor to elicit acute-phase protein production by murine hepatocytes. DESIGN: Experimental study. SETTING: Research laboratory at the DVA Medical Center, St Louis, Mo. CELLS STUDIED: Hepatocytes prepared from normal mice, from knockout mice deficient in IL-1 production due to loss of IL-1 converting enzyme, or from knockout mice deficient in the IL-1 p80 receptor. INTERVENTIONS: Cells were treated with lipopolysaccharide, a crude C difficile toxin extract, or purified C difficile toxins A or B for 24 hours in vitro, then radiolabeled with (35)S methionine. Newly synthesized acute-phase proteins were identified by electrophoresis and autoradiography. MAIN OUTCOME MEASURES: Synthesis of a 23-kd acute-phase protein in response to the various stimuli. RESULTS: Lipopolysaccharide, C difficile culture extract, and purified toxins A and B stimulated the synthesis of the 23-kd acute-phase protein by hepatocytes from normal mice and by hepatocytes from knockout mice deficient in the IL-1 converting enzyme. However, hepatocytes from knockout mice deficient in the IL-1 p80 receptor failed to produce this acute-phase protein when treated with the C difficile toxins, although they responded fully to lipopolysaccharide. CONCLUSIONS: Stimulation of acute-phase protein synthesis by C difficile toxins does not require IL-1 production, but does require a functioning IL-1 p80 receptor. This suggests that some of the actions of these toxins are mediated by this receptor. PMID- 11030884 TI - Absorbable mesh sling prevents radiation-induced bowel injury during "sandwich" chemoradiation for rectal cancer. AB - HYPOTHESIS: Absorbable mesh slings can prevent radiation-induced bowel injury when adjuvant pelvic radiotherapy is given in the early postoperative period. We hypothesized that the mesh sling technique is similarly effective during "sandwich" sequence adjuvant chemoradiation. DESIGN: Retrospective review. SETTING: Tertiary care comprehensive cancer center. PATIENTS: Nonrandomized series of 19 consecutive patients who underwent abdominoperineal resection and received postoperative sandwich sequence chemoradiation at Roswell Park Cancer Institute, Buffalo, NY, between January 1994 and September 1999. INTERVENTIONS: Twelve patients had an absorbable mesh sling placed at the completion of abdominoperineal resection. Seven patients did not have an absorbable mesh sling placed. MAIN OUTCOME MEASURES: Radiotherapy dose and gastrointestinal toxic effects. RESULTS: All 12 patients in the "mesh" group were able to receive full dose radiotherapy with tumor bed boost (total dose, 54 Gy, 11 patients; 59.4 Gy, 1 patient). Only 3 of 7 patients in the "no mesh" group were able to receive a tumor bed boost (total dose, 46.8 Gy, 1 patient; 50.4 Gy, 3 patients; 54 Gy, 3 patients). Acute gastrointestinal toxic effects were minimal in the mesh group (grade 1, 10 patients; grade 2, 2 patients) compared with the no mesh group (grade 2, 6 patients; grade 3, 1 patients). None of the patients in the mesh group have shown evidence of late gastrointestinal toxic effects. One patient in the no mesh group required surgery for complications of chronic radiation enteritis. CONCLUSIONS: The protective effects of an absorbable mesh sling extend beyond the life expectancy of the mesh itself. Sandwich sequence chemoradiation should not preclude the use of the mesh sling technique. PMID- 11030885 TI - Lymphatic involvement in early gastric cancer: prevalence and prognosis in France. AB - BACKGROUND: The prognosis of early gastric cancer (EGC) is considered better than that of invasive gastric carcinoma, with a 5-year survival rate of more than 90% after surgery. The prevalence of lymph node metastasis in EGC ranges from 8% to 20% and is associated with a poor prognosis. HYPOTHESIS: The main prognostic factor of EGC in patients in France is lymphatic involvement. DESIGN, SETTING, AND PATIENTS: From January 1979 to December 1988, 332 patients with EGC were operated on in 23 centers of 2 of the French Associations for Surgical Research. Clinical, pathological, and therapeutic data were reviewed, and the reckoning point was in June 1996. MAIN OUTCOME MEASURES: The cumulative 5- and 7-year specific survival rates of EGC with or without lymphatic involvement. RESULTS: The cumulative 5- and 7-year specific survival rates of 332 patients with EGC (mean follow-up time, 80 months), excluding both operative and unrelated mortality, were 92% and 87.5%, respectively. Thirty-four patients (10.2%) had metastatic lymphatic spread: 13 exclusively in the lymphatic vessels close to the tumor, 17 in at least 1 lymph node, and 4 in both the lymphatic vessels and nodes. The rate of lymph node involvement (regardless of lymphatic vessel involvement) correlated significantly with submucosal invasion (P =. 05) and histologic undifferentiation (P =.03). Lymphatic vessel involvement correlated positively with lymph node involvement (P =. 003). Since 5- and 7-year survival rates of the 13 patients with EGC who had lymphatic vessel involvement without lymph node involvement did not differ significantly from those of patients who had EGC with lymph node involvement (85% and 84% vs 72% and 63%, respectively [P =.42]), all patients with lymph node and/or lymphatic vessel involvement were considered unique. Prognosis was poorest in these patients according to both univariate analysis (94% for 298 without node or vessel involvement vs 78% for 34 with node and/or vessel involvement; P =.006) and multivariate analysis (P =.01). Submucosal invasion was a prognostic factor independent of lymphatic involvement (P =.05). Five- and 7-year survival rates did not differ when the group of 211 patients for whom less than 15 lymph nodes were retrieved were compared with those (n = 51) for whom 15 or more lymph nodes were retrieved (95.5% vs 92% and 95.5% vs 88%, respectively), whether according to univariate (P =.21) or multivariate (P =.31) analysis. CONCLUSIONS: Our results suggest that both lymph node and lymphatic vessel involvement are important prognostic factors in patients with EGC. Lymphadenectomy in EGC is important to identify the high-risk population for whom prognosis is worse. The extent of lymphadenectomy (at least 15 nodes) in these patients, however, does not alter prognosis. PMID- 11030886 TI - Isolated resection of segment 8 for liver tumors: a new approach for anatomical segmentectomy. AB - HYPOTHESIS: Isolated resection of segment 8 (the right anterosuperior liver segment) is one of the most difficult hepatectomies to perform because of the location of segment 8, the relation between section 8 and the main intrahepatic vessels, and the absence of any anatomical landmarks. The few reports that deal with isolated resection of section 8 generally describe the use of a deep wedge transparenchymal transection. DESIGN: Original surgical technique. PATIENTS AND METHODS: The proposed technique is based on the extraparenchymal isolation and temporary clamping of the right anterior artery and portal branches, causing ischemic demarcation on the liver surface, which corresponds to the anatomical borders of the right paramedian segments (5 and 8). The liver is widely transected along the main hepatic fissure; then the pedicles of segment 8 are selectively ligated inside the parenchyma, and the resection is accomplished. This technique was used in 10 patients: 5 with hepatocellular carcinoma on cirrhosis and 5 with liver metastases. RESULTS: The mean operation time was 253 minutes. Intraoperative blood loss was minimal in all cases, and 7 patients did not require blood transfusion. Slight complications developed in 3 patients, and there was no operative death. The mean hospital stay was 9.3 days. CONCLUSIONS: This operative procedure is safe and ensures a complete anatomical resection of segment 8. The wide opening of the liver parenchyma facilitates hemostasis and makes it possible to obtain a correct resection margin. This technique is recommended for limited metastatic lesions located in segment 8 or for hepatocellular carcinoma arising in a cirrhotic liver. PMID- 11030887 TI - Defining rural surgery. PMID- 11030888 TI - Surgical reminiscence: serendipity salvages disaster. PMID- 11030889 TI - Moments in surgical history: Charles Richard Drew. PMID- 11030890 TI - Annihilating cold dark matter AB - Structure formation with cold dark matter (CDM) predicts halos with a central density cusp, which are observationally disfavored. If CDM particles have an annihilation cross section sigmav approximately 10(-29)(m/GeV) cm(2), then annihilations will soften the cusps. We discuss plausible scenarios for avoiding the early Universe annihilation catastrophe that could result from such a large cross section. The predicted scaling of core density with halo mass depends upon the velocity dependence of sigmav, and s-wave annihilation leads to a core density nearly independent of halo mass, which seems consistent with observations. PMID- 11030891 TI - Quintessential adjustment of the cosmological constant AB - We construct a time dependent adjustment mechanism for the cosmological "constant" which could be at work in a late Friedmann-Robertson-Walker universe dominated by quintessence and matter. It makes use of a Brans-Dicke field that couples to the evolving standard-model vacuum energy density. Our explicit model possesses a stable late-time solution with a fixed ratio of matter and field energy densities. No fine-tuning of model parameters or initial conditions is required. PMID- 11030892 TI - Matrix theory interpretation of discrete light cone quantization string worldsheets AB - We study the null compactification of type-IIA string perturbation theory at finite temperature. We prove a theorem about Riemann surfaces establishing that the moduli spaces of infinite-momentum-frame superstring worldsheets are identical to those of branched-cover instantons in the matrix-string model conjectured to describe M theory. This means that the identification of string degrees of freedom in the matrix model proposed by Dijkgraaf, Verlinde, and Verlinde is correct and that its natural generalization produces the moduli space of Riemann surfaces at all orders in the genus expansion. PMID- 11030893 TI - Direct measurement of the W boson width in p&pmacr; collisions at radicals = 1.8 TeV AB - This Letter describes a direct measurement of the W boson total decay width, gamma(W), using the Collider Detector at Fermilab. The measurement uses an integrated luminosity of 90 pb(-1), collected during the 1994-1995 run of the Fermilab Tevatron p&pmacr; collider. The width is determined by normalizing predicted signal and background distributions to 49 844 W-->enu candidates and 21 806 W-->&mgr;nu candidates in the transverse-mass region M(T)<200 GeV and then fitting the predicted shape to the 438 electron events and 196 muon events in the high- M(T) region, 100 of a single mode radiation field can be prepared with arbitrarily high fidelity by interaction with a sequence of two level atoms, prepared in a suitable initial state. No final state measurement of the atoms is needed. PMID- 11030897 TI - Mesoscopic quantum coherence in an optical lattice AB - We observe the quantum coherent dynamics of atomic spinor wave packets in the double-well potentials of a far-off-resonance optical lattice. With appropriate initial conditions the system Rabi oscillates between the left and right localized states of the ground doublet, and at certain times the wave packet corresponds to a coherent superposition of these mesoscopically distinct quantum states. The atom/optical double-well potential is a flexible and powerful system for further study of quantum coherence, quantum control, and the quantum/classical transition. PMID- 11030898 TI - Information-theoretic stochastic resonance in noise-floor limited systems: the case for adding noise AB - We show that in systems whose output must compete with a noise source, stochastic resonance (maximization of output signal-noise separation as a nonmonotonic function of input noise strength) exists even when measured in terms of fundamental statistical measures and optimal detector performance. This is in contrast to the commonly considered scenario where, without the competing noise, the system (e.g., a driven, overdamped particle moving in a double well potential) is essentially invertible and optimal detector performance monotonically deteriorates with increasing input noise strength. PMID- 11030899 TI - Decoherence and the rate of entropy production in chaotic quantum systems AB - We show that for an open quantum system which is classically chaotic (a quartic double well with harmonic driving coupled to a sea of harmonic oscillators) the rate of entropy production has, as a function of time, two relevant regimes: For short times it is proportional to the diffusion coefficient (fixed by the system environment coupling strength). For longer times (but before equilibration) there is a regime where the entropy production rate is fixed by the Lyapunov exponent. The nature of the transition time between both regimes is investigated. PMID- 11030900 TI - From low-dimensional synchronous chaos to high-dimensional desynchronous spatiotemporal chaos in coupled systems AB - The dynamic behavior of coupled chaotic oscillators is investigated. For small coupling, chaotic state undergoes a transition from a spatially disordered phase to an ordered phase with an orientation symmetry breaking. For large coupling, a transition from full synchronization to partial synchronization with translation symmetry breaking is observed. Two bifurcation branches, one in-phase branch starting from synchronous chaos and the other antiphase branch bifurcated from spatially random chaos, are identified by varying coupling strength epsilon. Hysteresis, bistability, and first-order transitions between these two branches are observed. PMID- 11030901 TI - Experimental evidence of time-delay-induced death in coupled limit-cycle oscillators AB - Experimental observations of time-delay-induced amplitude death in two coupled nonlinear electronic circuits that are individually capable of exhibiting limit cycle oscillations are described. The existence of multiply connected death islands in the parameter space of coupling strength and time delay for coupled identical oscillators is established. The existence of such regions was predicted earlier on theoretical grounds [Phys. Rev. Lett. 80, 5109 (1998); Physica (Amsterdam) 129D, 15 (1999)]. The experiments also reveal the occurrence of multiple frequency states, frequency suppression of oscillations with increased time delay, and the onset of both in-phase and antiphase collective oscillations. PMID- 11030902 TI - 3D imaging: wave front sensing utilizing a birefringent crystal AB - Utilizing a rotatable thin birefringent crystal and two polarizers, the three dimensional (3D) wave front of any light wave can be measured conveniently. In first experiments phase-front distortions as small as 15 &mgr;m are detected with a dynamic range of 3 mm and a spatial resolution of 50 &mgr;m. Such a dynamic range and spatial resolution exceed the performance of conventional wave front sensors of, e.g., the Shack-Hartmann type, significantly. Furthermore, the new system is rather insensitive against mechanical instabilities in opposition to interferometric and holographic techniques. PMID- 11030903 TI - Signature of wave localization in the time dependence of a reflected pulse AB - The average power spectrum of a pulse reflected by a disordered medium embedded in an N-mode waveguide decays in time with a power law t(-p). We show that the exponent p increases from 3 / 2 to 2 after N2 scattering times, due to the onset of localization. We compare two methods to arrive at this result. The first method involves the analytic continuation to an imaginary absorption rate of a static scattering problem. The second method involves the solution of a Fokker Planck equation for the frequency dependent reflection matrix, by means of a mapping onto a problem in non-Hermitian quantum mechanics. PMID- 11030904 TI - Femtosecond X-Ray fluorescence AB - Using few-cycle-driven coherent laser harmonics, K-shell vacancies have been created in light elements, such as boron (E(B) = 188 eV) and carbon (E(B) = 284 eV), on a time scale of a few femtoseconds for the first time. The capability of detecting x-ray fluorescence excited by few-femtosecond radiation with an accuracy of the order of 1 eV paves the way for probing the evolution of the microscopic environment of selected atoms in chemical and biochemical reactions on previously inaccessible time scales (<100 fs) by tracing the temporal evolution of the "chemical shift" of peaks associated with inner-shell electronic transitions in time-resolved x-ray fluorescence and photoelectron spectra. PMID- 11030905 TI - Substrate interactions, effects of symmetry breaking, and convection in a 2D horizontally shaken granular system AB - We describe experiments on a horizontally shaken [x = Asin(omegat)] single layer of hard spheres rolling on a nearly horizontal surface. We identify a novel substrate-mediated convective flow which occurs when the system is tilted slightly so that the weak gravitational force, g-->(eff), acting on the particles is not parallel to the driving direction. As the shaking amplitude is increased, the system progresses through four regimes: solid-flat, solid-inclined, convective, and disordered. The control parameter is the driving velocity, Aomega, rather than the usual Aomega(2) of vertically shaken 3D systems. At the onset of convection, the critical velocity is V(c) approximately sqrt[2g(eff)d]. PMID- 11030906 TI - Experimental study of the multipolar vortex instability AB - The instability of a vortex subjected to a stationary dipolar or tripolar constraint is studied experimentally by using a rotating deformable cylinder on which two or three rollers are applied. As the Reynolds number and the aspect ratio of the cylinder are varied, different modes of instability are observed and their wavelength and frequency are compared to theoretical predictions. Secondary instability and cyclic breakup are also evidenced in the elliptic geometry. PMID- 11030907 TI - Subpicosecond electro-optic measurement of relativistic electron pulses AB - Time-resolved measurements of the transverse electric field associated with relativistic electron bunches are presented. Using an ultrafast electro-optic sensor close to the electron beam, the longitudinal profile of the electric field was measured with subpicosecond time resolution and without time-reversal ambiguity. Results are shown for two cases: inside the vacuum beam line in the presence of wake fields, and in air behind a beryllium window, effectively probing the near-field transition radiation. Especially in the latter case, reconstruction of the longitudinal electron bunch shape is straightforward. PMID- 11030908 TI - Momentum transport from nonlinear mode coupling of magnetic fluctuations AB - A cause of observed anomalous plasma momentum transport in a reversed-field pinch is determined experimentally. Magnetohydrodynamic theory predicts that nonlinear interactions involving triplets of tearing modes produce internal torques that redistribute momentum. Evidence for the nonlinear torque is acquired by detecting the correlation of momentum redistribution with the mode triplets, with the elimination of one of the modes in the triplet, and with the external driving of one of the modes. PMID- 11030909 TI - Subharmonic resonances in plasmas: exponential and superexponential growth of driven relativistic plasma waves AB - Subharmonic resonant beat-wave excitation of nonlinear relativistic plasma waves is studied analytically and in particle-in-cell simulations. We find that if the frequency separation of the lasers, Deltaomega, is 2omega(p) or 3omega(p) ( omega(p) is the plasma frequency), then plasma waves are still excited at omega(p) but they grow exponentially or superexponentially rather than secularly. Both of these subharmonic resonant instabilities saturate due to relativistic detuning. The analytical growth rates and saturation levels agree with the simulation results. PMID- 11030910 TI - Magnetic geometry and the confinement of electrically conducting plasmas AB - We develop an effective field theory approach to inspect the electromagnetic interactions in an electrically neutral plasma, with an equal number of negative and positive charge carriers. We argue that the static equilibrium configurations within the plasma are topologically stable solitons that describe knotted and linked flux tubes of helical magnetic fields. PMID- 11030911 TI - In situ diagnostics in plasmas of electronic-ground-state hydrogen molecules in high vibrational and rotational states by laser-induced fluorescence with vacuum ultraviolet radiation AB - Detection of excited electronic-ground-state hydrogen molecules with v(") up to 13 in a magnetic multipole plasma source was performed for the first time by laser-induced fluorescence with vacuum-ultraviolet radiation. The measurements are taken after fast shutoff of the discharge current. The rovibrationally excited molecules live longer than the plasma background light so that the fluorescence light can be detected with good signal-to-noise ratio. Absolute level populations are measured as well as decay times. The theoretically predicted suprathermal population of the vibrational distribution is clearly identified. The H- density is calculated on the basis of the measured populations and the measured electron energy distribution function. It is in excellent agreement with the H- density measured by photodetachment. PMID- 11030912 TI - Effective Co-generation of opposite and forward waves in cyclotron-resonance masers AB - A novel type of the frequency-tunable oscillator based on simultaneous generation of the forward and opposite waves at the same frequency, but at different cyclotron harmonics, is proposed. A spatially periodic helical electron beam allows for strong coupling of the waves. The opposite wave provides the broadband feedback and electron bunching, whereas the forward wave amplifies the arising signal and withdraws the rf power from the interaction region. A 15% efficiency and 5% frequency bandwidth have been achieved in the first experiment. PMID- 11030913 TI - Discovering new ordered phases of block copolymers AB - We propose a new and general method for discovering novel ordered phases of block copolymer melts. The method involves minimizing a free energy functional in an arbitrary unit cell with respect to the composition profile and the dimensions of the unit cell, without any prior assumption of the microphase symmetry. Varying the initial conditions allows to search for different stable and metastable structures. Application of this method to ABC star and linear triblock copolymers using an approximate free energy reveals new morphologies not yet observed in experiment. PMID- 11030914 TI - Quantitative NMR characterization of long-range chain dynamics prior to reptation: polyethylene-oxide AB - The thorough analysis of the transverse magnetic relaxation of protons, attached to highly entangled polyethylene-oxide chains in the melt, reveals two striking chain-length dependent properties; these are interpreted from the description (reminiscent of the Rouse model) of the long-range chain dynamics supposed to occur prior to the reptation motion. Experimental results are well matched by this specific NMR approach which accounts for the novel properties and provides the monomeric friction coefficient and the terminal relaxation time, over the molecular weight range 65K to 760K. PMID- 11030915 TI - Polyhedral units and network connectivity in calcium aluminosilicate glasses from high-energy X-Ray diffraction AB - Structure factors for Ca (x/2)Al xSi 1-xO (2) glasses (x = 0,0.25,0. 5,0.67) extended to a wave vector of magnitude Q = 40 A (-1) have been obtained by high energy x-ray diffraction. For the first time, it is possible to resolve the contributions of Si-O, Al-O, and Ca-O coordination polyhedra to the experimental atomic pair distribution functions. It has been found that the connectivity of Si/Al-O tetrahedral network decreases with increasing x due to the emerging of nonbridging oxygens located on Si-O tetrahedra. Calcium maintains a rather uniform coordination sphere for all values of x and so it plays a certain role in determining the glass structure. PMID- 11030916 TI - Origin of anomalous lattice expansion in oxide nanoparticles AB - Anomalous lattice expansions have been measured for the first time in monodisperse CeO2-x nanoparticles and in BaTiO3 single nanoparticles by electron diffraction. X-ray photoelectron spectroscopy studies on CeO2-x nanoparticles and ab initio computer simulation on BaTiO3 clusters show that the origin of expansion is the decrease of electrostatic force caused by valence reduction of Ce ions and the increase in ionicity of Ti ions, respectively. The lattice constant change of oxide (ionic) nanoparticles with the increase in ionicity would depend on the structure of the particles. Hence, first-principles calculations of large ionic clusters are indispensable. PMID- 11030917 TI - Melting of the rare earth metals and f-electron delocalization AB - Melting curves for Pr, Nd, Sm, Gd, and Y were measured in a diamond-anvil-cell to nearly 100 GPa and 4000 K. f-electron volume collapses are observed as triple points for Pr (24 GPa and 1400 K) and Gd (65 GPa and 3100 K). These pressures coincide with the volume collapses observed at room temperature. For Nd and Sm, the f-electron volume collapse has not been observed at room temperature but appears at approximately 2000-2500 K as a broad minimum in the melting curve, similar to that of Ce, near 50 GPa (Nd) and 70 GPa (Sm). The melting curve of Y goes smoothly along the entire rare earth sequence. PMID- 11030918 TI - Hole-burning experiments within glassy models with infinite range interactions AB - We reproduce the results of nonresonant spectral hole-burning experiments with glassy models with infinite-range interactions that generalize the mode-coupling approach to nonequilibrium situations. We show that an ac field modifies the integrated linear response and the correlation in a way that depends on the amplitude and frequency of the pumping field. We study the effect of the waiting and recovery times and the number of oscillations applied. This calculation will help discriminate which results can and which cannot be attributed to spatial heterogeneities in real systems. PMID- 11030919 TI - Testing the kibble-zurek scenario with annular josephson tunnel junctions AB - In parallel with Kibble's description of the onset of phase transitions in the early Universe, Zurek has provided a simple picture for the onset of phase transitions in condensed matter systems, supported by agreement with experiments in 3He and superconductors. We show how experiments with annular Josephson tunnel junctions can, and do, provide further support for this scenario. PMID- 11030920 TI - Semisuperfluidity of 3He in aerogel? AB - According to hydrodynamic, acoustic, and NMR studies the superfluid transition temperature of 3He in aerogel ( T(a)(c)) is significantly suppressed with respect to that of bulk 3He. We have found in the range of temperatures between T(c) and T(a)(c) a large and unexpected NMR satellite line attributable to the liquid inside the aerogel. We propose that this anomalous behavior of liquid 3He corresponds to a new type of superfluid ordering related to magnetic and possibly orbital coherence. PMID- 11030921 TI - Effect of branching on the fluid phase behavior of alkane monolayers AB - Configurational-bias Monte Carlo simulations in the grand canonical ensemble with histogram reweighting were used to obtain the vapor-liquid coexistence curves of three hexane isomers physisorbed on a flat gold substrate. Examination of the critical ordering operator distributions confirms that these systems exhibit critical behavior consistent with the 2D Ising universality class. The critical temperatures for 2,2-dimethylbutane, 2,3-dimethylbutane, and n-hexane were determined as T(2D)(c) = 195.0, 161.2, and 152.4 K, respectively. This is qualitatively different from the behavior observed for bulk (3D) fluids, where branched alkanes typically exhibit lower critical temperatures than their linear counterparts. PMID- 11030922 TI - Spin transport in interacting quantum wires and carbon nanotubes AB - We present a general formulation of spin-dependent transport through a clean one dimensional interacting quantum wire or carbon nanotube, connected to noncollinear ferromagnets via tunnel junctions. The low energy description of each junction is given by a conformally invariant boundary condition representing exchange coupling, in addition to a pair of electron tunneling operators. The effects of the exchange coupling are strongly enhanced by interactions, leading to a dramatic suppression of spin accumulation: a direct signature of spin-charge separation. Finally, backscattering induces nonequilibrium oscillations in the current-voltage relation. PMID- 11030923 TI - Covalent bonds in AlMnSi icosahedral quasicrystalline approximant AB - Electron density distributions were obtained using the maximum entropy method with synchrotron radiation powder data. In the metallic Al12Re, metallic bonding was observed for the icosahedral Al12 cluster with central Re atom. In the nonmetallic alpha-AlMnSi 1/1 approximant, covalent bonds were found in the electron density distribution of the Mackay icosahedral cluster without central atom. Rather than the Hume-Rothery mechanism, the covalency of Al (Si) icosahedron and that between Al (Si) and Mn atoms is considered to be the origin of the pseudogap and nonmetallic behavior of alpha-AlMnSi. PMID- 11030924 TI - Origin of surface conductivity in diamond AB - Hydrogen-terminated diamond exhibits a high surface conductivity (SC) that is commonly attributed to the direct action of hydrogen-related acceptors. We give experimental evidence that hydrogen is only a necessary requirement for SC; exposure to air is also essential. We propose a mechanism in which a redox reaction in an adsorbed water layer provides the electron sink for the subsurface hole accumulation layer. The model explains the experimental findings including the fact that hydrogenated diamond is unique among all semiconductors in this respect. PMID- 11030925 TI - Electronic transport in Y-junction carbon nanotubes AB - Electronic transport measurements were performed on Y-junction carbon nanotubes. These novel junctions contain a large diameter tube branched into smaller ones. Independent measurements using good quality contacts on both individual Y junctions and many in parallel show intrinsic nonlinear transport and reproducible rectifying behavior at room temperature. The results were modeled using classic interface physics for a junction with an abrupt change in band gap due to the change in tube diameter. These Y-junction tubes represent new heterojunctions for nanoelectronics. PMID- 11030926 TI - Andreev peaks and massive magnons in cuprate superconductor-normal-superconductor junctions AB - The projected SO(5) theory [pSO(5)] is used to resolve the puzzle of two distinct energy gaps in high T(c) superconductor-normal-superconductor junctions. Counter to the conventional theory of multiple Andreev reflections (MAR), the differential resistance peaks are associated with the antiferromagnetic resonance observed in neutron scattering, and not with Cooper pair breaking. The pSO(5) and MAR theories differ by the expected tunneling charges at the peaks. We propose that shot noise experiments could discriminate against the conventional interpretation. PMID- 11030927 TI - Intrinsic finite-size effects in the two-dimensional XY model with irrational frustration AB - This study investigates in detail the finite-size scaling of the two-dimensional irrationally frustrated XY model. By means of Monte Carlo simulations with entropic sampling, we examine the size dependence of the specific heat, and find remarkable deviation from the conventional finite-size scaling theory, which reveals novel intrinsic finite-size effects. Relaxation dynamics of the system is also considered, and, correspondingly, finite-size scaling of the relaxation time is examined, again giving evidence for the intrinsic finite-size effects and suggesting a zero-temperature glass transition. PMID- 11030928 TI - Measurement of vortex motion in a type-II superconductor: A novel use of the neutron spin-echo technique AB - We have used the neutron spin-echo technique to measure the small energy change of neutrons which are diffracted by a moving vortex lattice in a low-pinning Nb Ta superconducting sample. A transport current was passed in the mixed state to cause flux line movement. In the case of uniform motion, the flux velocity v(L) was given as expected by the values of electric and magnetic fields, via E = v(L)wedgeB. We show that with a nonuniformly moving vortex lattice, one can measure the dispersion of the velocities, opening up new possibilities for investigating moving vortex lines. PMID- 11030929 TI - Coulomb-Gas scaling law for a superconducting Bi(2+y)Sr(2-x-y)La(x)CuO(6+delta) thin films in magnetic fields AB - The electrical transport properties of epitaxial superconducting Bi(2+y)Sr(2-x y)La(x)CuO(6+delta) thin films have been studied in magnetic fields. Using a modified Coulomb-gas scaling law, we can fit all the magnetic field dependent low resistance data with a universal scaling curve, which allows us to determine a relation between the activation energy of the thermally activated flux flow resistance and the characteristic temperature scaling parameters. PMID- 11030930 TI - Ga NMR study of the local susceptibility in Kagome-based SrCr8Ga4O19: pseudogap and paramagnetic defects AB - We present the first Ga(4f ) NMR study of the Cr susceptibility in the archetype of kagome-based frustrated antiferromagnets, SrCr8Ga4O19. Our major finding is that the susceptibility of the frustrated lattice goes through a maximum around 50 K. Our data also support the existence of paramagnetic "clusters" of spins, responsible for the Curie behavior observed in the macroscopic susceptibility at low T. These results set novel features for the constantly debated physics of geometrically frustrated magnets. PMID- 11030931 TI - Observation of charge-density wave domains on the Cr(110) surface by low temperature scanning tunneling microscopy AB - We have studied the Cr(110) surface with low-temperature scanning tunneling microscopy at 6 to 145 K. For tunneling voltages below +/-150 mV we observe a surface charge-density wave (CDW) with a wavelength of 42 A and wave fronts aligned with the [001] in-plane direction. The observed wave pattern is identified as the surface projection of the bulk CDW's arising from the spin density wave ground state of Cr with the Q vector parallel to [100] and [010], respectively. The bulk CDW with Q parallel to the [001] in-plane direction appears, however, to be strongly suppressed at the Cr(110) surface. PMID- 11030932 TI - Low temperature spin dynamics of the geometrically frustrated antiferromagnetic garnet Gd3Ga5O12 AB - The low temperature spin dynamics of the geometrically frustrated antiferromagnet Gd 3Ga 5O (12) (GGG) have been investigated using muon spin relaxation. No evidence for static order is seen down to a temperature of 25 mK or a few percent of the Curie-Weiss temperature. Instead there is a linear decrease in the Gd spin fluctuation rate below 1 K which extrapolates to a small but finite value of 2 GHz at zero temperature. In terms of the spin fluctuations the system appears essentially to remain dynamic at low temperatures (T>0.02 K) and magnetic fields up to 1.8 T. PMID- 11030933 TI - Signature of electron-plasmon quantum kinetics in GaAs AB - We predict a carrier-density dependent oscillation, which is superimposed on the decay of the coherent control photon echo signal of a semiconductor. It reflects the oscillatory transfer of excitation back and forth between electrons and a mixed plasmon-phonon mode. This signature provides obvious and unique evidence for the finite duration of the interaction process, i.e., evidence for the collective Coulomb quantum kinetics. The theoretical predictions for the model semiconductor GaAs are reproduced in corresponding experiments. PMID- 11030934 TI - Interaction of excess electrons with water molecules at the early stage of laser induced plasma generation in water AB - Forward and backward stimulated Raman scattering (SRS) of OH stretching vibrations are measured when an intense 40 ps pulsed beam is focused into water. Characteristic features related to the interaction of OH groups with excess electrons generated by the strong laser irradiation are observed. The SRS spectra are indicative of the important role which excess electrons play in transient enhancement of the SRS at the initial stage of laser-induced plasma generation in water. The hydration structures around the excess electrons in water are also discussed. PMID- 11030935 TI - Dynamics of laser desorption and ablation of metals at the threshold on the femtosecond time scale AB - The dynamics of the laser-ablation (-desorption) process of metals (Al, Ag, Fe, and Ni) initiated by 30 fs laser pulses has been investigated by interferometric time-resolved pump-probe measurements. It is postulated that a sufficiently high density of hot electrons is essential for achieving desorption of metal ions. In addition, we have observed a new and unexpected behavior characterized by delayed ablation for a pump-probe beam delay in the range of several ps for Al, Ni, and Fe. This second peak is attributed to the development of a liquid surface layer developing after a few ps. Molecular dynamics simulations support this assumption. PMID- 11030936 TI - Dynamical generation of noiseless quantum subsystems AB - We combine dynamical decoupling and universal control methods for open quantum systems with coding procedures. By exploiting a general algebraic approach, we show how appropriate encodings of quantum states result in obtaining universal control over dynamically generated noise-protected subsystems with limited control resources. In particular, we provide a constructive scheme based on two body Hamiltonians for performing universal quantum computation over large noiseless spaces which can be engineered in the presence of arbitrary linear quantum noise. PMID- 11030937 TI - Validity of threshold-crossing analysis of symbolic dynamics from chaotic time series AB - A practical and popular technique to extract the symbolic dynamics from experimentally measured chaotic time series is the threshold-crossing method, by which an arbitrary partition is utilized for determining the symbols. We address to what extent the symbolic dynamics so obtained can faithfully represent the phase-space dynamics. Our principal result is that such a practice can lead to a severe misrepresentation of the dynamical system. The measured topological entropy is a Devil's staircase-like, but surprisingly nonmonotone, function of a parameter characterizing the amount of misplacement of the partition. PMID- 11030938 TI - Soap froths and crystal structures AB - We propose a physical mechanism to explain the crystal symmetries found in macromolecular and supramolecular micellar materials. We argue that the packing entropy of the hard micellar cores is frustrated by the entropic interaction of their brushlike coronas. The latter interaction is treated as a surface effect between neighboring Voronoi cells. The observed crystal structures correspond to the Kelvin and Weaire-Phelan minimal foams. We show that these structures are stable for reasonable areal entropy densities. PMID- 11030939 TI - Voronoi tessellation reveals the condensed matter character of folded proteins. AB - The packing geometry of amino acids in folded proteins is analyzed via a modified Voronoi tessellation method which distinguishes bulk and surface. From a statistical analysis of the Voronoi cells over 40 representative proteins, it appears that the packings are in average similar to random packings of hard spheres encountered in condensed matter physics, with a quite strong fivefold local symmetry. Moreover, the statistics permits one to establish a classification of amino acids in terms of increasing propensity to be buried in agreement with what is known from chemical considerations. PMID- 11030940 TI - Nonequilibrium phase transition in a model for social influence AB - We present extensive numerical simulations of the Axelrod's model for social influence, aimed at understanding the formation of cultural domains. This is a nonequilibrium model with short range interactions and a remarkably rich dynamical behavior. We study the phase diagram of the model and uncover a nonequilibrium phase transition separating an ordered (culturally polarized) phase from a disordered (culturally fragmented) one. The nature of the phase transition can be continuous or discontinuous depending on the model parameters. At the transition, the size of cultural regions is power-law distributed. PMID- 11030941 TI - Stability of the period-doubled core of the 90 degrees partial in silicon PMID- 11030942 TI - Comment on "Charged impurity-scattering-limited low-temperature resistivity of low-density silicon inversion layers" PMID- 11030943 TI - Das sarma and hwang reply: PMID- 11030944 TI - Comment on "Asymmetry of critical exponents in YBa(2)Cu(3)O(7-delta)" PMID- 11030945 TI - Comment on "Polarized raman study of aligned multiwalled carbon nanotubes" PMID- 11030946 TI - Rao et al. reply: PMID- 11030947 TI - Exact monte carlo method for continuum fermion systems AB - We offer a new proposal for the Monte Carlo treatment of many-fermion systems in continuous space. It is based upon diffusion Monte Carlo with significant modifications: correlated pairs of random walkers that carry opposite signs, different functions "guide" walkers of different signs, the Gaussians used for members of a pair are correlated, and walkers can cancel so as to conserve their expected future contributions. We report results for free-fermion systems and a fermion fluid with 14 3He atoms, where it proves stable and correct. Its computational complexity grows with particle number, but slowly enough to make interesting physics within the reach of contemporary computers. PMID- 11030948 TI - Robustness and diffusion of pointer states AB - Classical properties of an open quantum system emerge through its interaction with other degrees of freedom (decoherence). We treat the case where this interaction produces a Markovian master equation for the system. We derive the corresponding distinguished local basis (pointer basis) by three methods. The first demands that the pointer states mimic as closely as possible the local nonunitary evolution. The second demands that the local entropy production be minimal. The third imposes robustness on the inherent quantum and emerging classical uncertainties. All three methods lead to localized Gaussian pointer states, their formation and diffusion being governed by well-defined quantum Langevin equations. PMID- 11030949 TI - Fluctuation and relaxation properties of pulled fronts: A scenario for nonstandard kardar-parisi-zhang scaling AB - We argue that while fluctuating fronts propagating into an unstable state should be in the standard Kardar-Parisi-Zhang (KPZ) universality class when they are pushed, they should not when they are pulled: The 1/t velocity relaxation of deterministic pulled fronts makes it unlikely that the KPZ equation is their proper effective long-wavelength low-frequency theory. Simulations in 2D confirm the proposed scenario, and yield exponents beta approximately 0.29+/-0.01, zeta approximately 0.40+/-0.02 for fluctuating pulled fronts, instead of the (1+1)D KPZ values beta = 1/3, zeta = 1/2. Our value of beta is consistent with an earlier result of Riordan et al., and with a recent conjecture that the exponents are the (2+1)D KPZ values. PMID- 11030950 TI - Discrete periodic melting point observations for nanostructure ensembles AB - We report a study of the thermodynamic properties of indium clusters on a SiN (x) surface during the early stages of thin film growth using a sensitive nanocalorimetry technique. The measurements reveal the presence of abnormal discontinuities in the heat of melting below 100 degrees C. These discontinuities, for which temperature separation corresponds to a spatial periodicity equal to the thickness of an indium monolayer, are found to be related to the atomic "magic numbers," i.e., the number of atoms necessary to form a complete shell of atoms at particle surface. PMID- 11030951 TI - Generic isolated horizons and their applications AB - The notion of isolated horizons is extended to allow for distortion and rotation. Space-times containing a black hole, itself in equilibrium but possibly surrounded by radiation, satisfy these conditions. The framework has three types of applications: (i) it provides new tools to extract physics from strong field geometry; (ii) it leads to a generalization of the zeroth and first laws of black hole mechanics and sheds new light on the "origin" of the first law; and (iii) it serves as a point of departure for black hole entropy calculations in nonperturbative quantum gravity. PMID- 11030952 TI - Technique for direct eV-scale measurements of the Mu and tau neutrino masses using supernova neutrinos AB - Early black hole formation in a core-collapse supernova will abruptly truncate the neutrino fluxes. The sharp cutoff can be used to make model-independent time of-flight neutrino mass tests. Assuming a neutrino luminosity of 10(52) erg/s per flavor at cutoff and a distance of 10 kpc, Super-Kamiokande can detect an electron neutrino mass as small as 1.8 eV, and the proposed OMNIS detector can detect mu and tau neutrino masses as small as 6 eV. We present the first technique with direct sensitivity to eV-scale mu and tau neutrino masses. PMID- 11030953 TI - Natural chaotic inflation in supergravity AB - We propose a chaotic inflation model in supergravity. In the model the Kahler potential has a Nambu-Goldstone-type shift symmetry of the inflaton chiral multiplet which ensures the flatness of the inflaton potential beyond the Planck scale. We show that chaotic inflation naturally takes place by introducing a small breaking term of the shift symmetry in the superpotential. This may open a new branch of model building for inflationary cosmology in the framework of supergravity. PMID- 11030954 TI - Pion photo- and electroproduction and the partially conserved axial current AB - The relevance of the axial current for pion production processes off the nucleon with real or virtual photons is revisited. Employing the hypothesis of a partially conserved axial current (PCAC), it is shown that, when all of the relevant contributions are taken into account, PCAC does not provide any additional constraint for threshold production processes that goes beyond the Goldberger-Treiman relation. In particular, it is shown that pion electroproduction processes at threshold cannot be used to extract any information regarding the weak axial form factor. The relationships found in previous investigations are seen to be an accident of the approximations usually made in this context. PMID- 11030955 TI - Dynamic symmetries at the critical point AB - A new class of dynamic symmetries is introduced. It is suggested that an element of this class, associated with zeros of Bessel functions, be used to describe spectra of nuclei at or around the critical point of the U(5)-SO(6) shape phase transition, and, in general, spectra of systems undergoing a (second order) phase transition between the algebraic structures U(n-1) and SO(n). PMID- 11030956 TI - Evidence for a possible E(5) symmetry in 134Ba AB - Evidence for the first empirical example of a transitional dynamical symmetry at a critical point is discussed in the spectrum of 134Ba. The role of such classes of symmetries in nuclear structural evolution is also discussed. PMID- 11030957 TI - Caloric curves and energy fluctuations in the microcanonical liquid-Gas phase transition AB - In this paper we study a microcanonical lattice gas model with a constrained average volume. We show that the caloric curve explicitly depends on the considered transformation of the volume with the excitation energy and so does not bear direct information on the characteristics of the phase transition. Conversely, partial energy fluctuations are demonstrated to be a direct measure of the equation of state. Since the heat capacity has a negative branch in the phase transition region, the presence of abnormally large kinetic energy fluctuations is a signal of the liquid-gas phase transition. PMID- 11030958 TI - Multiple scattering, parton energy loss, and modified fragmentation functions in deeply inelastic eA scattering AB - Modified quark fragmentation functions in deeply inelastic eA collisions and their QCD evolution equations are derived for the first time in the framework of multiple parton scattering. Induced radiation gives rise to additional terms in the evolution equations and thus softens the modified quark fragmentation functions. The results in the next-leading-twist depend on both diagonal and off diagonal twist-four parton distributions and the combination of which clearly manifests the LPM interference pattern. The predicted modification depends quadratically on the nuclear size ( A2/3). Generalization to the case of hot QCD medium is also discussed. PMID- 11030959 TI - Observation of direct photons in central 158A GeV (208)P(208)b+Pb collisions AB - A measurement of direct photon production in 208Pb+208Pb collisions at 158A GeV has been carried out in the CERN WA98 experiment. The invariant yield of direct photons in central collisions is extracted as a function of transverse momentum in the interval 0.51.5 GeV/c. The result constitutes the first observation of direct photons in ultrarelativistic heavy ion collisions. It could be significant for diagnosis of quark-gluon-plasma formation. PMID- 11030960 TI - Observation of anti-stokes fluorescence cooling in thulium-doped glass AB - We report the first observation of anti-Stokes fluorescence cooling in a thulium doped solid with pump excitation at 1.82 &mgr;m and exist only over a narrow range of Ge coverages, 3.5-3.9 monolayers. PMID- 11030979 TI - Velocity fluctuations in a homogeneous 2D granular gas in steady state AB - We have measured the spectrum of velocity fluctuations in a granular system confined to a vertical plane and driven into a homogeneous, steady state by strong vertical vibration. The distribution of horizontal velocities is not Maxwell-Boltzmann and is given by P(v) = Cexp[-beta(|v|/sigma)(alpha)] where alpha = 1.55+/-0.1 at all frequencies and amplitudes investigated, and also for varying boundary conditions. The deviation from Maxwell-Boltzmann statistics occurs in the absence of spatial clustering and does not result from an inhomogeneous average over regions of varying local density. Surprisingly, P(v) has the same shape over a wide range of densities. PMID- 11030980 TI - Continuous wave observation of massive polariton redistribution by stimulated scattering in semiconductor microcavities AB - A massive redistribution of the polariton occupancy to two specific wave vectors, zero and approximately 3.9x10(4) cm(-1), is observed under conditions of continuous wave excitation of a semiconductor microcavity. The "condensation" of the polaritons to the two specific states arises from stimulated scattering at final state occupancies of order unity. The stimulation phenomena, arising due to the bosonic character of the polariton quasiparticles, occur for conditions of resonant excitation of the lower polariton branch. High energy nonresonant excitation, as in most previous work, instead leads to conventional lasing in the vertical cavity structure. PMID- 11030981 TI - Sn/Ge(111) surface charge-density-wave phase transition AB - Angle-resolved photoemission has been utilized to study the surface electronic structure of 1 / 3 monolayer of Sn on Ge(111) in both the room-temperature (sqrt[3]xsqrt[3] )R30 degrees phase and the low-temperature ( 3x3) charge-density wave phase. The results reveal a gap opening around the ( 3x3) Brillouin zone boundary, suggesting a Peierls-like transition despite the well-documented lack of Fermi nesting. A highly sensitive electronic response to doping by intrinsic surface defects is the cause for this unusual behavior, and a detailed calculation illustrates the origin of the ( 3x3) symmetry. PMID- 11030982 TI - Optical investigation of spin-wave excitations in fractional quantum hall states and of interaction between composite fermions AB - We demonstrate that the temperature dependence of the electron spin polarization for the fractional states nu = 1/3 and nu = 2/3 displays activated behavior. This study enables the first measurement of the fractional quantum Hall spin-flip gaps. They are found to be systematically larger in comparison with the gaps simultaneously measured in transport. For nu = 1/3 and nu = 1/2, these spin-flip gaps allow the determination of the composite fermion interaction energy. This energy is investigated as a function of the finite width of the 2D channel. PMID- 11030983 TI - Tunneling density of states of high T(c) superconductors: d-wave BCS model versus SU(2) slave-boson model AB - Motivated by recent experimental measurements of the tunneling characteristics of high T(c) materials using scanning tunneling spectroscopy, we have calculated the I-V and differential conductance curves in the superconducting state at zero temperature. Comparing BCS-like d-wave pairing and the SU(2) slave-boson approach, we find that the slave-boson model can explain the asymmetric background observed in experiments. The slave-boson model also predicts that the height of the conductance peak relative to the background is proportional to the hole doping concentration x, at least for underdoped samples. We also observe the absence of the van Hove singularity, and comment on possible implications. PMID- 11030985 TI - Evidence for nodal quasiparticles in electron-doped cuprates from penetration depth measurements AB - The in-plane magnetic penetration depth, lambda(T), was measured down to 0.4 K in single crystals of electron-doped superconductors, Pr(1.85)Ce(0.15)CuO(4-delta) (PCCO) and Nd(1.85)Ce(0.15)CuO(4-delta) (NCCO). In PCCO, the superfluid density varies as T2 from 0.025 up to roughly 0.3T/T(c) suggestive of a d-wave state with impurities. In NCCO, lambda(T) shows a pronounced upturn for T<4 K due to the paramagnetic contribution of Nd3+ ions. Fits to an s-wave order parameter over the standard BCS range (T/T(c) = 0.32) limit any gap to less than Delta(min)(0)/T(c) = 0.57 in NCCO. For PCCO, the absence of paramagnetism permits a lower temperature fit and yields an upper limit of Delta(min)(0)/T(c) = 0.2. PMID- 11030984 TI - Microwave electrodynamics of electron-doped cuprate superconductors AB - We report microwave cavity perturbation measurements of the temperature dependence of the penetration depth, lambda(T), and conductivity, sigma(T) of Pr(2-x)Ce(x)CuO(4-delta) (PCCO) crystals, as well as parallel-plate resonator measurements of lambda(T) in PCCO thin films. Penetration depth measurements are also presented for a Nd(2-x)Ce(x)CuO(4-delta) (NCCO) crystal. We find that Deltalambda(T) has a power-law behavior for T1 GHz NMR in unstable magnetic fields. AB - Resistive or hybrid magnets can achieve substantially higher fields than those available in superconducting magnets, but their spatial homogeneity and temporal stability are unacceptable for high-resolution NMR. We show that modern stabilization and shimming technology, combined with detection of intermolecular zero-quantum coherences (iZQCs), can remove almost all of the effects of inhomogeneity and drifts, while retaining chemical shift differences and J couplings. In a 25-T electromagnet (1 kHz/s drift, 3 kHz linewidth over 1 cm(3)), iZQC detection removes >99% of the remaining inhomogeneity, to generate the first high-resolution liquid-state NMR spectra acquired at >1 GHz. PMID- 11030994 TI - Correlated and uncorrelated regions in heart-rate fluctuations during sleep. AB - Healthy sleep consists of several stages: deep sleep, light sleep, and rapid eye movement (REM) sleep. Here we show that these sleep stages can be characterized and distinguished by correlations of heart rates separated by n beats. Using the detrended fluctuation analysis (DFA) up to fourth order we find that long-range correlations reminiscent to the wake phase are present only in the REM phase. In the non-REM phases, the heart rates are uncorrelated above the typical breathing cycle time, pointing to a random regulation of the heartbeat during non-REM sleep. PMID- 11030995 TI - Functional stochastic resonance in the human brain: noise induced sensitization of baroreflex system. AB - We demonstrate experimentally that noise can enhance the homeostatic function in the human blood pressure regulatory system. The results show that the compensatory heart rate response to the weak periodic signal introduced at the venous blood pressure receptor is optimized by adding noise to the arterial blood pressure receptor. We conclude that this functional stochastic resonance most likely results from the interaction of noise with signal in the brain stem, where the neuronal inputs from these two different receptors first join together. PMID- 11030996 TI - Structure and dynamics of reactive intermediates in reaction mechanisms. sigma- and pi-complexes in electrophilic aromatic substitutions PMID- 11030997 TI - Intramolecular carbostannation reaction of active methine compounds with an unactivated C-C pi-bond mediated by SnCl(4)-Et(3)N AB - In the presence of SnCl(4) and Et(3)N, intramolecular carbostannation reaction of various active methine compounds having an unactivated alkenyl, alkynyl, or allenyl group proceeds in good yields with complete regioselectivity. The subsequent reaction of the resulting Sn intermediates with some electrophiles provides functionalized cyclopentane and cyclohexane derivatives. PMID- 11030998 TI - Stable dimeric aromatic cation-radicals. Structural and spectral characterization of through-space charge delocalization AB - The spontaneous assembly of aromatic cation-radicals (D(+)(*)()) with the parent donor (D) to afford the paramagnetic dimer (D)(2)(+)(*)() is accompanied by a dramatic color change. For example, spectral (UV-vis and ESR) and X-ray crystal structure analyses establish the molecular association of octamethylbiphenylene cation-radical with its neutral counterpart to produce the mixed-valence or dimeric cation-radical in which the positive charge is completely delocalized over both aromatic moieties. The use of the sterically hindered cation-radicals confirms the new spectral or charge-resonance (CR) band to result in dimeric cation-radicals in which the intermolecular separation occurs at an optimum distance allowed by van der Waals contacts. The striking similarities between the classical donor/acceptor (EDA) complexes and the dimeric cation-radicals (D)(2)(+)(*)() (both in terms of the geometrical requirement as well as the appearance of new absorption bands) suggest that the latter can be considered as particular examples of Mulliken's charge-transfer complexes in which the positive charge is completely (equally) delocalized over both donor (D) and acceptor (D(+)(*)()). PMID- 11031000 TI - Copper-mediated synthesis of phthalocyanino-fused Dehydro AB - Cyclooligomeric phthalocyanines with the special feature of having a dehydroannulene core are described. Compounds 4a,b and 5a,b were prepared by an oxidative coupling mediated by copper from the corresponding diethynyl unsymmetrically substituted phthalocyanines 8a,b, which are easily accessible from the corresponding diiodo compounds 6a,b by a palladium-copper-catalyzed coupling. Appropriate conditions to selectively form cyclodimers and cyclotrimers using Glaser and Eglinton methodologies have been attempted. In this way, two compounds, 4a and 5b, each representative of a kind of oligomers, could be appropriately isolated and characterized by MALDI-MS. The UV-vis spectra of compounds 4a and 5b show a split Q-band shifted to the red with respect to the corresponding precursor 8a,b. This can be interpreted, in light of previous results, as a consequence of intramolecular electronic coupling between the Pc subunits. Extensive formation of aggregates in chloroform can be also inferred from the spectra. No significant differences could be observed in the spectra by comparison with that of the open-chain analogue 11, most probably due to a saturation phenomenon of the optical properties already pointed out in the literature for other alkynyl-linked open-chain dimers. PMID- 11031001 TI - Incorporation of stable organic radicals of the tris(2,4, 6 trichlorophenyl)methyl radical series to pyrrole units as models for semiconducting polymers with high spin multiplicity. 1. AB - The condensation reactions between (4-amino-2,6-dichlorophenyl)bis(2, 4,6 trichlorophenyl)methyl radical and acetylacetone or 1, 4-bis(5-methyl-2-thienyl) 1,4-butanedione yield [2,6-dichloro-4-(2, 5-dimethyl-1-pyrrolyl)phenyl]bis(2,4,6 trichlorophenyl)methyl radical (3(*)()) and [2,6-dichloro-4-[2, 5-bis(5-methyl-2 thienyl)-1-pyrrolyl]phenyl]bis(2,4, 6-trichlorophenyl)methyl radical (4(*)()), respectively. EPR studies of both radicals 3(*)() and 4(*)() in CH(2)Cl(2) solution suggest a weak electron delocalization with coupling constant values of 1.25 and 1.30 G, respectively, with the six aromatic hydrogens. Their electrochemical behavior was analyzed by cyclic voltammetry. Both radicals show reversible reduction processes at E degrees = -0.69 V and -0.61 V versus SSCE, respectively, and anodic peak potentials at E(p)(a) = 1.10 and 0.72 V, respectively, versus SSCE at a scan rate (nu) of 200 mV s(-)(1), being reversible for radical 4(*)(). X-ray analysis of radical 3(*)() shows a high value (65 degrees ) of the dihedral angle between the 2,5-dimethylpyrrolidyl moiety and the phenyl ring. Smooth oxidation of radical 4(*)() in CH(2)Cl(2) containing trifluoroacetic acid gives an ionic diradical species with a weak electron interaction (|D/hc| = 0.0047 cm(-)(1)). A Curie plot of the Deltam(s)() = +/-2 signal intensity versus the inverse of the absolute temperature in the range between 4 and 70 K suggests a triplet or a nearly degenerate singlet-triplet ground state. PMID- 11030999 TI - Honulactones: new bishomoscalarane sesterterpenes from the Indonesian sponge Strepsichordaia aliena. AB - From the dichloromethane/2-propanol (1:1) extract of the Indonesian marine sponge Strepsichordaia aliena, twelve new 20, 24-bishomoscalarane sesterterpenes, honulactones A-L (1-12) were isolated. Molecular structures were secured by spectroscopic methods, accurate mass measurements, and X-ray analysis. Honulactones A (1), B (2), C (3), and D (4) exhibit cytotoxycity against P-388, A 549, HT-29, and MEL-28 (IC(50) 1 microg/mL). PMID- 11031002 TI - Cyano-, nitro-, and alkoxycarbonyl-activated observable stable enols of carboxylic acid amides AB - A search for the enol structures of several amides YY'CHCONHPh with Y,Y' = electron-withdrawing groups (EWGs) was conducted. When Y = CN, Y' = CO(2)Me the solid structure is that of the enol (8b) MeO(2)CC(CN)=C(OH)NHPh, whereas in solution the NMR spectrum indicate the presence of both the amide MeO(2)CCH(CN)CONHPh (8a) and 8b. When Y = NO(2), Y' = CO(2)Et the main compound in CDCl(3) is the amide, but <10% of enol(s), presumably EtO(2)CC(NO(2))=C(OH)NHPh (9b), are also present. When Y = COEt, Y' = CO(2)Me or Y = COMe, Y' = CO(2)Et (10 and 11) enolization in solution and of 11 also in the solid state occurs at the carbonyl rather than at the ester site. With Y = Y' = CN a rapid exchange between the amide (NC)(2)CHCONHPh (12a) and a tautomer, presumably the enol, take place in several solvents on the NMR time scale. With YY' = barbituric acid moiety the species in DMSO-d(6) is an enol of an amide although which CONH group enolizes is unknown. B3LYP/6-31G calculations showed that the enol (NC)(2)C=C(OH)NH(2) (13b) is more stable by DeltaG of 0.4 kcal/mol than (NC)(2)CHCONH(2) (13a) due to a combination of stabilization of 13b and destabilization of 13a and both are much more stable than the hydroxyimine and ketene imine tautomers. The effect of Y,Y' and the solvent on the relative stabilization of enols of amides is discussed. PMID- 11031003 TI - How borane reagents based on alpha-pinene control stereoselectivity in the reductions of carbonyl groups with different stereogenic elements. 3. The effect of the relative size and conformation of the carbonyl substituents on the stereoselectivity of the Ipc(2)BCl, Eap(2)BCl, B-t-Bu-IpcBCl, and B-t-Bu-EapBCl reagents. A semiempirical study AB - The reductions of a more elaborate prostereogenic nonchiral and chiral ketones with the representative borane reagents derived from (+)-alpha-pinene confirm that the overall stereoselectivity, similarly to the previously studied reductions of benzaldehyde and acetophenone, is also controlled, early along the reaction coordinate, by the structure of the borane reagent. The stereoselectivity reflects the energy differences among syn-1, 3-interactions of the smallest of the three boron substituents in the reacting borane conformation with the two carbonyl substituents in the substrate. For the B-alkyl-IpcBCl reagents and the R(S)-CO-R(L) substrates, these are the interactions of the B-Cl with the C-R(S) and C-R(L) bonds. To minimize developing intermolecular syn-1,3 interactions with the reagent's B-Cl bond during the reaction, in the conformationally mobile R(S)-CO-R(L) systems, the R(L) and R(S) groups seek to adopt a more reactive conformation even at the expense of the unfavorable intramolecular interactions. PMID- 11031004 TI - Reaction of singlet oxygen with trans-4-propenylanisole. Formation Of AB - We report the effects of added acid in the reaction of singlet oxygen with trans 4-propenylanisole (1). We provide evidence that solvent acidity modifies the behavior of the transient intermediates. Relative to reactions in aprotic solvent, enhanced dioxetane concentrations are observed in MeOH and in nonprotic solvents with acid. We suggest a new mechanism that invokes a proton transfer from MeOH and benzoic acid to perepoxide (2) and zwitterion (3) intermediates. PMID- 11031005 TI - Catalysis of ester aminolysis by cyclodextrins. The reaction of alkylamines with p-nitrophenyl alkanoates AB - The effects of four cyclodextrins (alpha-CD, beta-CD, hydroxypropyl-beta-CD, and gamma-CD) on the aminolysis of p-nitrophenyl alkanoates (acetate to heptanoate) by primary amines (n-propyl to n-octyl, isobutyl, isopentyl, cyclopentyl, cyclohexyl, benzyl) in aqueous solution have been investigated. Rate constants for amine attack on the free and CD-bound esters (k(N) and k(cN)) have ratios (k(cN)/k(N)) varying from 0.08 (retardation) to 180 (catalysis). For the kinetically equivalent process of free ester reacting with CD-bound amine (k(Nc)), the ratios k(Nc)/k(N) vary from 0.2 to 28. Either way, there is evidence of catalysis in some cases and retardation in others. Changes in reactivity parameters with structure indicate more than one mode of transition state binding to the CDs. Short esters react with short alkylamines by attack of free amine on the ester bound by its aryl group, but for longer amines, free ester reacts with CD-bound amine. Reaction of long esters with long amines, which is catalyzed by beta-CD and gamma-CD, involves inclusion of the alkylamino group and possibly the ester acyl group. The larger cavity of gamma-CD may allow the inclusion of the ester aryl group, as well as the alkylamino group, in the transition state. Reaction between an ester bound to the CD by its acyl group and free amine appears not to be important. PMID- 11031006 TI - Photolysis of bromo- and chloro-substituted benzyl derivatives. Competition between ionic and radical pathways AB - Photolysis of 1-fluoro-2-halo-1,2-diphenylethanes was studied in solutions of tetrahydrofuran, acetonitrile, and cyclohexane. The effect of free radical inhibitor and metal hydrides on products formation as well as their ratio was analyzed to elucidate the reaction pathway. In the first step homolytic C-X bond cleavage occurs from a single excited state, resulting in a biradical pair. Further reaction path depends on the type of the halogen bonded and on the solvent polarity. Electron transfer within the radical pair cage is apparently more rapid for bromides than for chlorides and is opposite as expected on the basis of the relative electronegativities of chlorine and bromine. As radicals approach each other, they fall into ionic or radical product channels. This is influenced by solvent polarity, resulting in the larger yield of ionic products in the case of acetonitrile as in the case of less polar cyclohexane. PMID- 11031007 TI - Epoxidation of alpha,beta-unsaturated ketones using hydrogen peroxide in the presence of basic hydrotalcite catalysts AB - The basic layered hydrotalcites have been used as catalysts for the epoxidation of alpha,beta-unsaturated ketones in heterogeneous reaction media using hydrogen peroxide as an oxidant. A wide variety of alpha,beta-unsaturated ketones were oxidized to the corresponding epoxyketones in excellent yields under mild reaction conditions. For example, 2-cyclohexen-1-one gave 2,3-epoxycyclohexanone in 91% yield at 40 degrees C for 5 h with high efficiency in hydrogen peroxide. The catalytic activity of the hydrotalcites increased as the basicity of their surfaces increased. In the case of the epoxidation of less reactive substrates, adding a cationic surfactant such as n-dodecyltrimethylammonium bromide (DTMAB) to the above oxidation system accelerated the epoxidation reaction. These hydrotalcite catalysts were easily separated from the reaction mixture and were reusable. PMID- 11031008 TI - Synthesis of gallic acid: Cu(2+)-mediated oxidation of 3-dehydroshikimic acid. AB - With the elaboration of high-yielding, high-titer syntheses of 3-dehydroshikimic acid from glucose using recombinant Escherichia coli, oxidation of this hydroaromatic becomes a potential route for synthesis of gallic acid. Conversion of 3-dehydroshikimic acid into gallic acid likely proceeds via initial enolization of an alpha-hydroxycarbonyl and oxidation of the resulting enediol. 3 Dehydroshikimate enolization in water was catalyzed by inorganic phosphate while Zn(2+) was used to catalyze enolization in acetic acid. Enediol oxidation employed Cu(2+) as either the stoichiometric oxidant or as a catalyst in the presence of a cooxidant. Gallic acid was produced in a yield of 36% when 3 dehydroshikimic acid in phosphate-buffered water reacted for 35 h with H2O2 and catalytic amounts of CuSO(4). 3-Dehydroshikimate-containing, phosphate-buffered culture supernatants reacted with stoichiometric amounts of CuCO(3)Cu(OH)(2) and Cu(x)(H(3-x)(PO4)(2) to give gallic acid in yields of 51% in 5 h and 43% in 12 h, respectively. Solutions of 3-dehydroshikimic acid in acetic acid reacted with stoichiometric amounts of Cu(OAc)(2) to afford a 74% yield of gallic acid in 36 h. Acetic acid solutions of 3-dehydroshikimic acid could also be oxidized by air using catalytic quantities of Cu(OAc)(2). ZnO accelerated these oxidations leading to a 67% yield of gallic acid in 4 h when an acetic acid solution of 3 dehydroshikimic acid was reacted with O(2) and a catalytic amount of Cu(OAc)(2). PMID- 11031009 TI - Ab initio and density functional evaluations of the molecular conformations of beta-caryophyllene and 6-hydroxycaryophyllene. AB - The four conformations of beta-caryophyllene (alphaalpha, alphabeta, betaalpha, and betabeta) were investigated ab initio at the 6-31G/HF and MP2 levels and additionally with density functional methods (B3LYP/6-31G), as it concerns their relative thermodynamic stabilities. The alphaalpha is predicted to be the most stable geometry, in agreement with low-temperature NMR measurements. In the case of 6-hydroxycaryophyllene, the alphaalpha is still the most stable conformation when the configuration at C-6 is S, but when the configuration is reversed to R the betabeta geometry becomes the most stable one. This is again in agreement with NMR data. On the other hand, for both molecules the AM1 semiempirical model Hamiltonian fails to predict the alphaalpha as a low-energy geometry, mainly due to an incorrect description of the cyclobutane ring puckering. The interconversion paths among the different minima are also analyzed and discussed. The solvent effect (either chloroform or water) on the stability of the different conformers of beta-caryophyllene and 6-hydroxycaryophyllene was studied in the polarizable continuum model framework. PMID- 11031010 TI - Synthesis, solid-state structures, and aggregation motifs of phosphines and phosphine oxides bearing one 2-pyridone ring AB - Three phosphines and their corresponding oxides bearing one 2-pyridone ring and two benzene rings were synthesized. Their single-crystal X-ray analyses exhibited three kinds of molecular aggregation: bimolecular aggregates, chiral one dimensional structures, and achiral one-dimensional structures. In the bimolecular aggregate of (2-oxo-1, 2-dihydro-x-pyridyl)diphenylphosphines (x = 3: 2a and 6: 2c), cyclic dimers that are derived from two 2-pyridone rings are observed. In contrast, (2-oxo-1,2-dihydro-5-pyridyl)diphenylphosphine (2b) molecules form a chiral one-dimensional chain via intermolecular hydrogen bonding. In the case of phosphine oxides, their oxygen always acts as a hydrogen acceptor of the hydrogen bonding. Thus, (2-oxo-1,2-dihydro-x pyridyl)diphenylphosphine oxides (x = 3: 3a and 5: 3b) form hydrogen bonds intermolecularly between the oxygen atom on the phosphoryl group and the hydrogen atom on nitrogen to construct a chiral or an achiral one-dimensional chain. Interestingly, (2-oxo-1,2-dihydro-6-pyridyl)diphenylphosphine oxide (3c) exists as a 2-hydroxypyridine form (enol form) in a crystalline state, and intermolecular hydrogen bonds between the phosphoryl oxygen and the hydroxy proton construct an achiral one-dimensional chain. PMID- 11031011 TI - Radical cyclization in heterocycle synthesis. 11.(1) A novel synthesis of alpha,beta-disubstituted gamma-lactones via sulfanyl radical addition-cyclization using hydroximates as a tether AB - A combination of sulfanyl radical addition-cyclization of dienes connected with hydroximates and subsequent conversion of the resulting cyclic hydroximate to the lactones provides a novel method for the construction of alpha,beta-disubstituted gamma-lactones. Upon treatment with thiophenol in the presence of AIBN, dienes connected with hydroximates smoothly underwent sulfanyl radical addition cyclization to give cyclic cis- and trans-hydroximates. Hydrolysis of cyclic hydroximates gave the desired cis- and trans-lactones in high yield. This method was successfully applied to the practical synthesis of (+/-)-oxo-parabenzlactone. PMID- 11031012 TI - Combinatorial liquid-phase synthesis of AB - A new method for the synthesis of [1,4]oxazepin-7-ones from readily available aldehydes and alpha-amino alcohols was developed using the Baylis-Hillman reaction as the key step. To determine the scope and limitations of the method, a mixture library was synthesized from six aldehydes and six alpha-amino alcohols on the soluble polymer support poly(ethylene glycol) 5000 monomethyl ether (MeOPEG) via split synthesis and analyzed by GC-EIMS. Those oxazepines that were formed predominantly were resynthesized in a parallel synthesis and fully characterized. Thus, we have shown that split synthesis on MeOPEG can be an efficient method to rapidly screen the substrate spectrum of a newly developed reaction sequence. PMID- 11031013 TI - A four-step enzymatic cascade for the one-pot synthesis of non-natural carbohydrates from glycerol. AB - A total of four enzymatic steps were combined, in a one-pot reaction, to synthesize carbohydrates starting from glycerol. First, phosphorylation of glycerol by reaction with pyrophosphate in the presence of phytase at pH 4.0 in 95% glycerol afforded racemic glycerol-3-phosphate in 100% yield. The L enantiomer of the latter underwent selective aerobic oxidation to dihydroxyacetone phosphate (DHAP) at pH 7.5 in the presence of glycerolphosphate oxidase (GPO) and catalase. Subsequently, fructose-1,6-bisphosphate aldolase catalyzed the aldol reaction of DHAP with butanal. Finally, dephosphorylation of the aldol adduct was mediated by phytase at pH 4 affording 5-deoxy-5-ethyl-D xylulose in 57% yield from L-glycerol-3-phosphate. The phytase on/off-switch by pH was the key to controlling phosphorylation and dephosphorylation. PMID- 11031014 TI - Selective palladium-catalyzed cocyclotrimerization of arynes with dimethyl acetylenedicarboxylate: A versatile method for the synthesis of polycyclic aromatic hydrocarbons AB - Benzyne (1a) and the substituted derivatives 4,5-difluorobenzyne (1b) and 3 methoxybenzyne (2) undergo chemoselective palladium-catalyzed [2 + 2 + 2] cocyclotrimerization with dimethyl acetylenedicarboxylate (DMAD) to afford the corresponding phenanthrenes and/or naphthalenes. The major products are phenanthrenes if Pd(PPh(3))(4) is used as the catalyst, naphthalenes if Pd(2)(dba)(3) is used. When the method is applied to polycyclic arynes 3-6, which are generated from the corresponding o-trimethylsilylaryl triflates, the same reactivity pattern is observed: the reaction can be selectively directed either toward the cocyclization of one molecule of aryne and two molecules of alkyne or to the reaction of two molecules of aryne with one molecule of alkyne, by appropriate choice of the palladium catalyst. The synthesis of polycyclic aromatic compounds 33-39 using this methodology is reported. PMID- 11031015 TI - Preparation of diynes via selective bisalkynylation of zirconacycles AB - Reaction of alkynyl halides with in situ prepared zirconacyclopentanes, pentenes, and -pentadienes in the presence of CuCl under mild reaction conditions afforded alkynes or diynes. Control of the reaction conditions selectively afforded monoalkynylation products of zirconacycles. Reaction of zirconacycles with 2 equiv of alkynyl halides resulted in the formation of diynes. Selective monoalkynylation of zirconacycle with an alkynyl halide, followed by reaction with a different alkynyl halide, afforded unsymmetrical diynes. Bisalkynylation product of zirconacyclopentadiene was gradually converted into a tricyclic compound. PMID- 11031016 TI - On the Z-E photoisomerization of chiral 2-pentenoate esters: stationary irradiations, laser-flash photolysis studies, and theoretical calculations. AB - Chiral pentenoates 1-3 in both Z and E isomeric forms underwent stationary irradiations in several solvents and in the presence of different photosensitizers. The photostationary-state ratio has been determined for each Z/E couple showing a predominance of the thermodynamically more stable isomer for 1 and 3. Moreover, transient species were generated by pulsed laser excitation and detected by their characteristic ultraviolet absorptions, being the first time that enoate-originated triplets are detected. Stern-Volmer quenching studies afforded a quantitative measure for the efficiency of the photosensitization processes induced by benzophenone or acetophenone and allowed the determination of the corresponding quenching rate constants. Density functional calculations permitted the determination of the geometries and the energies of the diastereomeric excited states. Two diastereomeric orthogonal and two diastereomeric planar structures result as a consequence of the presence of a chiral substituent. The orthogonal triplets are the energy minima in all cases, whereas the planar triplets are the transition states linking these orthogonal structures, the corresponding energy barriers being 8-10 kcal mol(-1) for enoates 1-3. The computed S(0) to T(1) excitation energies show a trend which is consistent with the quenching rate constants. On the other hand, the triplet lifetimes determined for 1 and 2 are unusually long (1-20 micros) if compared with the data already described for several enones, in the range of nanoseconds. This fact has been rationalized from calculations of spin-orbit coupling at several points of the T(1) potential energy surface. This coupling is maximum for structures with a torsional angle close to 45 degrees, which are 4-5 kcal mol(-1) above the minima of T(1). Calculations done on the hypothetical aldehyde 4 and methyl vinyl ketone show much lower energy barriers, thus accounting for the shorter lifetimes reported for enone triplets. PMID- 11031017 TI - Highly enantioselective approach to indolizidines: preparation of (+)-(1S,8aS)-1 hydroxyindolizidine and (-)-slaframine. AB - A highly stereoselective approach to (-)-slaframine and its probable biosynthetic precursor (+)-(1S,8aS)-1-hydroxyindolizidine has been developed based on a diastereofacially selective cycloaddition of dichloroketene with a chiral dienol ether. PMID- 11031018 TI - Novel ferrocenyl polyene derivatives and their binding to unmodified cyclodextrins AB - Two new ferrocene derivatives, 7-ferrocenyl-2,4,6-heptatrienal (1) and 7 ferrocenyl-2,4,6-heptatrienol (2), were synthesized and characterized. These two compounds possess a rigid triene chain conjugated to one of the cyclopentadienyl rings of the ferrocene residue, and as a result, they exhibit very stiff structures. The electronic absorption and electrochemical properties of these compounds were utilized to investigate their host-guest binding interactions with the receptors alpha-cyclodextrin (alpha-CD) and beta-cyclodextrin (beta-CD) in aqueous solution. From electronic absorption measurements binding constants in the range 790-12900 M(-)(1) were obtained; beta-CD formed more stable complexes than alpha-CD with both guests. Electrochemical measurements suggest some degree of site selectivity in the complexation processes, with beta-CD binding preferentially to the ferrocene moiety while alpha-CD interacts with the unsaturated chain. PMID- 11031019 TI - Synthesis of new aromatic (C17-C20)-locked side-chain analogues of calcitriol (1alpha,25-dihydroxyvitamin D(3)). AB - The synthesis of four new analogues of calcitriol (1alpha, 25-(OH)(2)-D(3)) possessing aromatic and conjugated double bond units at the side chain are described. The triene system is introduced using the Lythgoe-Hoffmann La Roche convergent Wittig-Horner approach. The key steps in the preparation of the requisite upper fragments are the introduction of the side chain with the E conjugated aromatic system and its photochemical conversion to the Z counterpart. PMID- 11031020 TI - Practical and highly enantioselective ring opening of cyclic meso-anhydrides mediated by cinchona alkaloids. AB - The cinchona alkaloid-mediated opening of prochiral cyclic anhydrides in the presence of methanol leading to optically active hemiesters is described. Very structurally diverse anhydrides are converted into their corresponding methyl monoesters, and either enantiomer can be obtained with up to 99% ee by using quinine or quinidine as directing additive. After the reaction, the alkaloids can be recovered almost quantitatively and reused without loss of enantioselectivity. Additionally, a catalytic protocol which permits the substoichiometric use of quinidine in the presence of easily accessible pentamethylpiperidine (pempidine) is presented. PMID- 11031021 TI - Oxazolopiperidin-2-ones as type II' beta-turn mimetics: synthesis and conformational analysis. AB - We describe a straightforward synthesis of 9-substituted 3 aminooxazolidinopiperidin-2-ones 4. Some derivatives were prepared for use in peptide synthesis as rigidified surrogates of the Ala-Pro dipeptide. Analysis of the amide derivatives 14 by NMR experiments and molecular mechanics/dynamics calculations shows that the major isomer 14a has a stronger propensity than the minor isomer 14b to adopt beta-turn conformations, and the calculations indicate that in water 14a adopts a stable betaII' turn conformation. PMID- 11031022 TI - Stereoselective 1,3-dipolar cycloadditions of a chiral nitrone derived from erythrulose. An experimental and DFT theoretical study AB - We have investigated several 1,3-dipolar cycloadditions of a chiral nitrone prepared from L-erythrulose. While cycloadditions to carbon-carbon multiple bonds of dipolarophiles such as ethyl acrylate, ethyl propiolate, or dimethyl acetylenedicarboxylate were poorly stereoselective, reaction with acrylonitrile provided predominantly one diastereomeric adduct. Furthermore, the regioselectivity exhibited by the two structurally similar dipolarophiles ethyl acrylate and ethyl propiolate was found to be opposite. The molecular mechanisms of these cycloadditions have thus been investigated by means of density functional theory (DFT) methods with the B3LYP functional and the 6-31G and 6 31+G basis sets. A simplified achiral version of nitrone 1 as the dipole, and methyl propiolate or acrylonitrile as the dipolarophiles, were chosen as computational models. The cycloadditions have been shown to take place through one-step pathways in which the C-C and C-O sigma bonds are formed in a nonsynchronous way. For the reaction with methyl propiolate, DFT calculations predict the experimentally observed meta regioselectivity. For the reaction with acrylonitrile, however, the predicted regioselectivity has been found to depend on the computational level used. The calculations further indicate the exo approach to be energetically favored in the case of the latter dipolarophile, in agreement with experimental findings. The main reason for this is the steric repulsion between the nitrile function and one of the methyl groups on the nitrone that progressively develops in the alternative endo approach. PMID- 11031023 TI - A combination of tandem amine-exchange/heterocyclization and biaryl coupling reactions for the straightforward preparation of phenanthro[9,10-d]pyrazoles. AB - The tandem amine-exchange/heterocyclization of enaminoketones is successfully applied to the regioselective preparation of a series of new 4,5-diarylpyrazoles by reaction of phenylhydrazine and several 3-N,N-(dimethylamino)-1,2 diarylpropenones. The comparison of a vast array of biaryl coupling procedures provides general, complementary approaches to new phenanthro[9,10-d]pyrazoles. The most efficient procedures for this final step in the construction of the tetracyclic system are based on a Stille-type tandem stannylation-biaryl coupling of o,o'-dihalogenated diarylpyrazoles and an hypervalent iodine-mediated nonphenolic oxidative coupling of nonhalogenated precursors. PMID- 11031024 TI - Total synthesis of (+)-zaragozic acid C. AB - A total synthesis of (+)-zaragozic acid C is described. Key features of the synthesis are the use of a double Sharpless asymmetric dihydroxylation reaction of diene 6 to control stereochemistry at four contiguous stereocenters from C3 to C6; the introduction of the C1-side chain by reaction between the anion derived from the dithiane monosulfoxide 27 and the core aldehyde 12; a high yielding, acid-mediated simultaneous acetonide deprotection-dithiane removal-ketalization procedure leading exclusively to the 2, 8-dioxabicyclo[3.2.1]octane core 34; and a novel triple oxidation procedure allowing installation of the tricarboxylic acid. PMID- 11031025 TI - Atropisomeric amides as chiral ligands: using (-)-sparteine-directed enantioselective silylation to control the conformation of a stereogenic axis AB - An enantiomerically pure (1-trimethylsilyl)ethyl group, constructed by a (-) sparteine-directed enantioselective quench of a laterally lithiated tertiary aromatic amide, exerts powerful thermodynamic control over the conformation of the adjacent tertiary amide substituent. Ortholithiation and functionalization of the amide in the 6-position allows the single amide conformer to be trapped as an enantiomerically and diastereoisomerically pure amide atropisomer. Protodesilylation of the amide gives functionalized atropisomeric amides with a stereogenic axis of single absolute configuration, whose barriers to racemization have been determined by polarimetry. Enantiomerically pure amides bearing phosphine substituents are effective ligands in a Pd-catalyzed allylic substitution reaction-the first use of a nonbiaryl atropisomer as a chiral ligand and give products with 90% ee. The rate of racemization of the phosphine substituted amide is powerfully influenced by the presence of palladium. PMID- 11031026 TI - Asymmetric PTC C-alkylation catalyzed by chiral derivatives of tartaric acid and aminophenols. Synthesis Of (R)- and (S)-alpha-methyl amino acids AB - A new type of efficient chiral catalyst has been elaborated for asymmetric C alkylation of CH acids under PTC conditions. Sodium alkoxides formed from chiral derivatives of tartaric acid and aminophenols (TADDOL's 2a-e and NOBIN's 3a-h) can be used as chiral catalysts in the enantioselective alkylation, as exemplified by the reaction of Schiff's bases 1a-e derived from alanine esters and benzaldehydes with active alkyl halides. Acid-catalyzed hydrolysis of the products formed in the reaction afforded (R)-alpha-methylphenylalanine, (R)-alpha naphthylmethylalanine, and (R)-alpha-allylalanine in 61-93% yields and with ee 69 93%. The procedure could be successfully scaled up to 6 g of substrate 1b. When (S,S)-TADDOL or (R)-NOBIN are used, the (S)-amino acids are formed. A mechanism rationalizing the observed features of the reaction has been suggested. PMID- 11031027 TI - Aryl H-phosphonates. 12. Synthetic and (31)P NMR studies on the preparation of nucleoside H-phosphonothioate and nucleoside H-phosphonodithioate monoesters. AB - Transformation of nucleoside H-phosphonate monoesters into the corresponding H phosphonothioate and H-phosphonodithioate derivatives and possible side-reactions that may accompany this process were studied using (31)P NMR spectroscopy. These provided new insight into a possible mechanism involved in this transformation and constituted the basis for development of efficient methods for the preparation of nucleoside H-phosphonothioate and nucleoside H-phosphonodithioate monoesters using readily available H-phosphonate monoesters as starting materials. PMID- 11031028 TI - Relationship between the configurations of 2-phenyltetrahydrothiophenium 1 methylides and their rearrangement products AB - trans-2-Phenyltetrahydrothiophenium 1-methylide (trans-3), which is generated by fluoride ion-induced desilylation of trans-2-phenyl-1 [(trimethylsilyl)methyl]tetrahydrothiophenium salt (trans-2), gave a mixture of 1,4,5,10a-tetrahydro-3H-2-benzothiocine (4) ([2,3]sigmatropic rearrangement product) and 4-methylsulfanyl-1-phenyl-1-butene (5) (Hofmann elimination product). Ylide trans-3 cannot undergo [2,3]sigmatropic rearrangement because the ylide-carbon is too far from the phenyl group, and trans-3 would instead isomerize to cis-3. In this paper, we discuss the mechanism of the isomerization of trans-3 to cis-3. PMID- 11031029 TI - Total synthesis of (+)-quassin. AB - A total synthesis of (+)-quassin from naturally occurring (S)-(+)-carvone is described. The total number of steps was 28, and the overall yield was about 2.6%. The synthetic strategy for the construction of the tetracyclic carbon framework was based on a C-->ABC-->ABCD ring annulation sequence, involving an aldol reaction, an intramolecular Diels-Alder reaction, and an intramolecular acylation as the key steps. Subsequent functionalization of ring A and ring C then afforded the target (+)-quassin. PMID- 11031030 TI - New approach to aphidicolin and total asymmetric synthesis of unnatural (11R)-(-) 8-epi-11-hydroxyaphidicolin by tandem transannular Diels-Alder/aldol reactions. AB - The 8-epiaphidicolane skeleton (3) was formed in one key reaction by highly diastereoselective tandem transannular Diels-Alder (TADA)-aldol reactions from the trans-trans-cis trienic macrocycle (4). The unnatural derivative (11R)-(-)-8 epi-11-hydroxyaphidicolin (2) was thus constructed, and an original solution to the C16 functionalization problem of many aphidicolin (1) syntheses is presented. PMID- 11031032 TI - Diastereoselective radical hydrogenation of alpha-(1-Hydroxyalkyl)vinyl sulfoxides and sulfones controlled by intramolecular hydrogen bonding AB - The reaction of (S)-alpha-(1-hydroxyalkyl)vinyl sulfoxides (S)-5 with alkyl radicals and tributyltin hydride gave the addition-hydrogenation products with high diastereoselectivity, whereas the reaction with (R)-alpha-(1 hydroxyalkyl)vinyl sulfoxides (R)-5 resulted in complete recovery of the starting sulfoxides. Stereoselective intramolecular hydrogen bonding between the hydroxy group and the diastereotopic sulfonyl oxygen led to high diastereoselectivity in the radical reaction of alpha-(1-hydroxyethyl)vinyl sulfone 12. An important role of intramolecular hydrogen bonding on the diastereoselectivity as well as the reactivity toward alkyl radicals is discussed. PMID- 11031031 TI - Synthesis of 2',3'-dideoxy-2'-trifluoromethylnucleosides from alpha trifluoromethyl-alpha,beta-unsaturated ester. AB - The treatment of alpha-bromo-alpha,beta-unsaturated esters 2 with FSO(2)CF(2)CO(2)Me and CuI in DMF/HMPA constitutes a new synthetic scheme for the preparation of alpha-trifluoromethyl-alpha, beta-unsaturated esters 3. The trifluoromethylation of (Z)/(E)-ethyl 3-[(S)-2,2-dimethyl-1,3-dioxolan-4-yl]-2 bromo-2-propenoate (2e), which is derived from 1-(R)-glyceraldehyde acetonide, yields the key intermediate alpha-trifluoromethyl-alpha,beta-unsaturated esters 3e. This is transformed into anomeric acetates 8a and 8b and is used for the synthesis of a number of 2', 3'-dideoxy-2'-trifluoromethylnucleosides. PMID- 11031033 TI - Chiral alpha-substituted carbonyls and alcohols from the S(N)2' displacement of cuprates on chiral carbonates: An alternative to the alkylation of chiral enolates AB - A highly stereoselective sequence of reactions, based on the anti-selective S(N)2' addition of cuprates to allylic carbonates, transforms alkynes or alkenyl halides into carbonyls having alpha-chiral centers. The method, which uses menthone as a chiral auxiliary, is a useful alternative to the alkylation of chiral enolates with the added advantage of allowing for the "alkylation" of sec- and tert-alkyl and aryl groups. PMID- 11031034 TI - Reductions with lithium in low molecular weight amines and ethylenediamine AB - Reductions of several types of compounds with lithium and ethylenediamine using low molecular weight amines as solvent are described. In all cases 1 mol of ethylenediamine or N, N'-dimethylethylenediamine per gram-atom of lithium was used. In some cases it was beneficial to add an alcohol as a proton donor. These reaction conditions were applied to the debenzylation of N-benzylamide and lactams which are refractory to hydrogenolysis with hydrogen and a catalyst. N Benzylpilolactam 2, synthesized from pilocarpine hydrochloride in refluxing benzylamine, was debenzylated in good yield using 10 gram-atoms of lithium per mole (10 Li/mol) of 2 in n-propylamine. The debenzylation of N-benzyl-N methyldecanoic acid amide, 4 (6 Li/mol), in t-butylamine/N, N' dimethylethylenediamine gave N-methyldecanoic acid amide 6 in 70% yield. Alternatively, reduction of 4 (7 Li/mol) in t-butanol/n propylamine/ethylenediamine gave n-decanal 12 in 36% yield. Using the same conditions, thioanisole, 1-adamantane-p-toluenesulfonamide, and 1-adamantane methyl p-toluenesulfonate were reduced with 3, 7, and 7.2 Li/mol of compound to give thiophenol (74%), adamantamine (91%), and 1-adamantane methanol (75%), respectively. In this solvent system naphthalene and 3-methyl-2-cyclohexene-1-one were reduced to isotetralin (74%) and 3-methyl cyclohexanone (quantitative) with 5 and 2.2 Li/mol of starting compound, respectively. Oximes and O-methyloximes were reduced to their corresponding amines using 5 and 8 Li/mol of compound, respectively. Anisole was also reduced to 1-methoxy-1,4-cyclohexadiene with 2.5 Li/mol of anisole. Undecanenitrile was reduced to undecylamine with 8.6 Li/mol. Additionally, a base-catalyzed formation of imidazolines from a nitrile and ethylenediamine was also explored. PMID- 11031035 TI - Molecular design of crown ethers. 21.(1,2) synthesis of novel double-armed benzo 15-crown-5 lariats and their complexation thermodynamics with light lanthanoid nitrates in acetonitrile AB - Three new disubstituted benzo-15-crown-5 derivatives (3-5) have been synthesized from 4',5'-bis(bromomethyl)benzo-15-crown-5 (2) and the corresponding alkanols in the presence of Na(2)S(2), and their complexation thermodynamics with light lanthanoid(III) nitrates (La-Gd) have been studied in anhydrous acetonitrile at 25 degrees C. Plots of K(S) against the reciprocal ionic diameter of lanthanoid exhibited monotonically declining pattern for the parent benzo-15-crown-5 (1) and 3 but showed a characteristic peak at Ce(3+) for 4 and 5. It is interesting to note that the simple extension of the alkyl side chains in 4 and 5 can alter the cation selectivity profiles of 1 and 3. Possessing two 2-oxapropyl groups, 3 gave a comparable K(S) for La(3+) but a significantly decreased K(S) for Ce(3+) compared with the corresponding values for 1, thus exhibiting an exceptionally high La(3+)/Ce(3+) selectivity of 11. Thermodynamically, the complexation of lanthanoid perchlorates with 1 is absolutely entropy-driven in acetonitrile, while the complexation of lanthanoid nitrates with 3-5 is primarily driven by exothermic enthalpy changes with accompanying moderate entropic gain or small entropic loss. PMID- 11031036 TI - A concise formal total synthesis of mappicine and nothapodytine B via an intramolecular hetero Diels-Alder reaction. AB - A six-step formal total synthesis of a natural alkaloid, mappicine (3), has been achieved. The highlight of our synthetic strategy is an intramolecular hetero Diels-Alder reaction that was used for the construction of the CD ring system of mappicine (3). In addition, it was demonstrated that the Sonogashira coupling reaction of 2-chloro-3-hydroxymethylquinoline (8c) with trimethylsilylacetylene proceeded at room temperature in excellent yield. PMID- 11031037 TI - Lipophilicity of the nitrophenols AB - The lipophilicity of the nitrophenols, expressed as a water-solvent partition coefficient, P, has been investigated using the solvation equation, log P = c + eE + sS + aA + bB + vV. It is shown that this equation accounts quantitatively for lipophilicity in a selection of water-solvent systems, viz: octanol, 1,2 dichloroethane, and cyclohexane. In the latter two systems, the major factor in the increased lipophilicity of 2-nitrophenol over 3- and 4-nitrophenol is the lack of hydrogen bond acidity of 2-nitrophenol. The water-octanol system differs in that the a coefficient is effectively zero, so that hydrogen bond acidity of solutes plays no part, and the three mononitrophenols then have similar lipophilicities. The dinitrophenols and picric acid are similarly discussed. The hydrogen bond acidity of 2,3-dinitrophenol (0.67) is very much larger than that of 2,4- or 2,5-dinitrophenol (0.09 and 0. 11), indicating a very much reduced internal hydrogen bonding. A similar but much smaller effect occurs with 2,6 dinitrophenol (A = 0. 17). Picric acid has a moderate hydrogen bond acidity (0.46) so that the phenolic OH is still available for external hydrogen bonding. These results are confirmed by ab initio calculations which show that 2,3- and 2,6-dinitrophenol and picric acid are significantly distorted away from planarity, which apparently disrupts their internal hydrogen bonding. PMID- 11031038 TI - Radical and anionic response of N-(Bromomethanesulfonyl)-substituted alpha,alpha' bridged piperidine substrates AB - The N-(bromomethanesulfonyl) azabicyclic ketones 2a-c and their exo-methylene analogues 1a-c were prepared. Our examination of the radical-induced behavior of the latter triad provided experimental evidence for the propensity of the b and c systems to engage in 7-endo cyclization. For 1a, only reductive debromination was observed. In no instance was ring closure by the 6-exo radical mode seen. As concerns ketones 2a-c, all three showed a remarkable facility for engaging in intramolecular S(N)2 displacement in the presence of KHMDS. Yields at the mid-80% level were realized irrespective of the value of n. Molecular mechanics calculations of the Monte Carlo type were performed on several conformational isomers and product classes in an effort to provide support for the explanatory conclusions offered as rationale for the collective experimental observations. PMID- 11031039 TI - Rhodium(II)-catalyzed cyclization of amido diazo carbonyl compounds. AB - A series of acyclic diazo ketoamides were prepared from N-benzoyl-N alkylaminopropanoic acids and were treated with a catalytic amount of rhodium(II) acetate. The resultant carbenoids underwent facile cyclization onto the neighboring amide carbonyl oxygen atom to generate seven-membered carbonyl ylide dipoles. Subsequent collapse of the dipoles with charge dissipation produce bicyclic epoxides which undergo further reorganization to give substituted 5 hydroxydihydropyridones in good yield. Depending on the nature of the substituent groups, it was possible to trap some of the initially formed carbonyl ylide dipoles with a reactive dipolarophile such as DMAD. In other cases, cyclization of the dipole to the epoxide is much faster than bimolecular trapping. A related cyclization/rearrangement sequence occurred when diazo ketoamides derived from the cyclic pyrrolidone and piperidone ring systems were subjected to catalytic quantities of Rh(II) acetate. With these systems, exclusive O-cyclization of the amido group onto the carbenoid center occurs to generate a seven-ring carbonyl ylide dipole. Starting materials are easily prepared, and the cascade sequence proceeds in good yield and does not require special precautions. The overall procedure represents an efficient one-pot approach toward the synthesis of various indolizidine and quinolizidine ring systems. PMID- 11031040 TI - Conformational selectivity in the diels-alder cycloaddition: predictions for reactions of s-trans-1,3-butadiene AB - Diels-Alder cycloaddition of s-trans-1,3-butadiene (1) should yield trans cyclohexene (7), just as reaction of the s-cis conformer gives cis-cyclohexene (9). Investigation of this long-overlooked process with Hartree-Fock, Moller Plesset, CASSCF, and DFT methods yielded in every case a C(2)-symmetric concerted transition state. At the B3LYP/6-31G (+ZPVE) level, this structure is predicted to be 42.6 kcal/mol above reactants, while the overall reaction is endothermic by 16.7 kcal/mol. A stepwise diradical process has been studied by UBLYP/6-31G theory and found to have barriers of 35.5 and 17.7 kcal/mol for the two steps. Spin correction lowers these values to 30.1 and 13.0 kcal/mol. The barrier to pi bond rotation in cis-cyclohexene (9) is predicted (B3LYP theory) to be 62.4 kcal/mol, with trans-cyclohexene (7) lying 53.3 kcal/mol above cis isomer 9. Results suggest that pi-bond isomerization and concerted reaction may provide competitive routes for Diels-Alder cycloreversion. It is concluded that full understanding of the Diels-Alder reaction requires consideration of both conformers of 1,3-butadiene. PMID- 11031041 TI - Synthesis, resolution, and absolute stereochemistry of (1R, 2S)-(+)-cis-1 methoxycarbonyl-2-methylcyclobutane AB - Racemic cis-1-methoxycarbonyl-2-methylcyclobutane uncontaminated with the trans isomer was prepared efficiently in five steps; the corresponding amides from (R) (-)-phenylglycinol were separated. An X-ray crystallographic structure determination of the amide from (+)-cis-1-methoxycarbonyl-2-methylcyclobutane showed that it was of (1R,2S) absolute stereochemistry, a revision of configurational assignment. PMID- 11031042 TI - Quantitative analyses of the seven isomeric 3,4- and 3, 6-dimethylcyclohexenes by gas chromatography AB - Quantitative analyses of mixtures of the seven isomeric 3,4- and 3, 6 dimethylcyclohexenes have been achieved by gas chromatography. Correlations of structure and absolute stereochemistry with elution order have been made rigorously with the aid of authentic optically active samples all derived from (3R)-methylcyclohexanone. PMID- 11031043 TI - Photoinduced electron-transfer reactions with quinolinic and trimellitic acid imides: experiments and spin density calculations(1) AB - The regioselectivity of photoinduced electron-transfer (PET) reactions of unsymmetrical phthalimides is controlled by the spin density distribution of the intermediate radical anions. ROHF ab initio calculations were found to be most suitable for atomic spin density analysis. Intramolecular PET reactions of quinolinic acid imides were studied with the potassium butyrate and hexanoate 1a,b and a cysteine derivative 3. The photocyclizations products 2a,b and 4 were formed with moderate regioselectivities (68:32, 57:43, and 81:19) showing preferential ortho cyclization. The intermolecular reaction of potassium propionate and potassium isobutyrate with N-methylquinolinic acid imide (5) yielded as addition products the dihydropyrrolo[3,4-b]pyridines 6a,b with slight ortho regioselectivity (55:45). In contrast to these low regioselectivities, the PET reaction of potassium propionate with the methyl ester of N-methyltrimellitic acid imide (9) yielded solely the para addition product 10. Likewise, the intramolecular photoreaction of the cysteine derivative 7 gave a 75:25 (para/meta) mixture of regioisomeric cyclization products 8. The regioselectivity originates from donor-acceptor interactions prior to electron transfer and differences in spin densities in the corresponding imide radical anions. The results of DFT and ab initio calculations for the radical anions of the quinolinic acid imide (11(*)(-)) and the methyl ester of trimellitic acid imide (12(*)(-))( )()were in agreement with the latter assumption: spin densities in 11(*)(-) were higher for the imido ortho carbon atoms (indicating preferential ortho coupling); for 12(*)(-) the spin densities were higher for the imido para carbon atoms (indicating preferential para coupling). These correlations became more significant when the additional spin densities at the carbonyl oxygen and the adjacent carbon atoms were taken into account. The cyclization selectivities for 2, 4, and 8 deviate from the intermolecular examples probably because of ground-state and solvent effects. PMID- 11031045 TI - Quantum yields of the initiation step and chain propagation turnovers in S(RN)1 reactions: photostimulated reaction of 1-iodo-2-methyl-2-phenyl propane with carbanions in DMSO AB - Neophyl radicals were generated by photoinduced electron transfer (PET) from a suitable donor to the neophyl iodide (1, 1-iodo-2-methyl-2-phenylpropane). The PET reaction of 1 with the enolate anion of cyclohexenone (2) afforded mainly the reduction products tert-butylbenzene (5) and the rearranged isobutylbenzene (6), arising from hydrogen abstraction of the neophyl radical (15) and the rearranged radical 16 intermediates, respectively. The photostimulated reaction of 1 with 2 in the presence of di-tert-butylnitroxide, as a radical trap, afforded adduct 10 in 57% yield. The photoinduced reaction of the enolate anion of acetophenone (3) with 1 gave the substitution products 11 (50%) and 12 (16%), which arise from the coupling of 3 with radicals 15 and 16, respectively. The rate constant obtained for the addition of anion 3 to radical 15 was 1.2 x 10(5) M(-)(1) s(-)(1), by the use of the rearrangement of this radical as a clock reaction. The anion of nitromethane (4) was almost unreactive at the initiation step, but in the presence of 2 under irradiation, it gave high yields (67%) of the substitution product 13 and only 2% of the rearranged product 14. When the ratio of 4 to 1 was diminished, it was possible to observe both substitution products 13 and 14 in 16% and 6.4% yields, respectively. These last results allowed us to estimate the coupling rate constant of neophyl radicals 15 with anion 4 to be at least of the order of 10(6) M(-)(1) s(-)(1). Although the overall quantum yield determined (lambda = 350 nm) for the studied reactions is below 1, the chain lengths (Phi(propagation)) for the reaction of 1 with anions 3 and 4 are 127 and 2, respectively. PMID- 11031044 TI - Synthesis of benzo-fused 1-azabicyclo[m.n.0]alkanes via the Schmidt reaction: a formal synthesis of gephyrotoxin. AB - The intramolecular capture of benzocyclobutyl, benzocyclopentyl, and benzocyclohexyl carbocations 7 by azides produces spirocyclic aminodiazonium ions 8, which undergo 1,2-C-to-N rearrangement with loss of dinitrogen to produce benzo-fused iminium ions resulting from either aryl (9) or alkyl (10) migration to the electron-deficient nitrogen atom. Reduction of the iminium ions affords regioisomeric benzo-fused 1-azabicyclo[m.n.0]alkanes, e.g., benzopyrrolizidines, benzoindolizidines, benzoquinolizidines, or perhydrobenzo[f]pyrrolo[1,2 a]azepines in two regioisomeric versions, anilines (e.g., 11-14) and benzylic amines (e.g., 15-18), the result of aryl and alkyl migrations, respectively. Generally, aryl migration is preferred, despite modeling that shows that the lowest energy aminodiazonium ions are those where the departing dinitrogen is preferentially antiperiplanar to the migrating alkyl group rather than the aryl group. The utility of this methodology was illustrated by a formal synthesis of the alkaloid gephyrotoxin 4. A dependence on the efficiency and regioselectivity of the Schmidt reaction upon subtle changes in the structure of the cation precursor was observed, necessitating the exploration of a variety of substrates. Fortunately, these materials were easily made. Ultimately, the azido-alkene 81 bearing a 2-bromoethyl side-chain was useful for the Schmidt reaction, producing the known benzo-fused indolizidine 49, which had been transformed by Ito et al. into gephyrotoxin 4. The synthesis of 49 required nine steps (five purifications) from commercially available 4-methoxy-1-indanone 60 and proceeded in 22% overall yield. PMID- 11031046 TI - Consequences of affinity in heterogeneous catalytic reactions: highly chemoselective hydrogenolysis of iodoarenes AB - The catalytic hydrodeiodination reaction using molecular hydrogen and Pd/C has been revisited. It is shown, for the first time, that the chemoselectivity of this reaction is controlled by the high affinity of the iodinated compound for the catalyst. This reaction is compatible with most easily reducible functional groups (nitro, aldehyde, olefin, etc.). Using this reaction, the first general method for tritium labeling of 3-(trifluoromethyl)-3-phenyldiazirine is described. PMID- 11031047 TI - Annulation strategies for benzo[b]fluorene synthesis: efficient routes to the kinafluorenone and WS-5995 antibiotics. AB - Intramolecular palladium-mediated arylation approaches to benzo[b]fluorenes have been investigated. The methodology has been applied in a short synthesis of tri-O methylkinafluorenone, providing an effective alternative to Friedel-Crafts-based approaches. During the course of this work, an acid-promoted quinolactonization of naphthoquinones was also developed, providing direct access to either ortho or para isomers as desired. Application of this methodology in syntheses of the antibiotics WS-5995A, WS-5995C, and functional analogues was demonstrated, and antitumoral activity of this class was determined. PMID- 11031049 TI - Metalation of a 3,5-dichloro-tertiary benzamide. An unusual regioselectivity observation PMID- 11031048 TI - Yanuthones: novel metabolites from a marine isolate of Aspergillus niger. PMID- 11031050 TI - Halogenation of 4-phenyl-3-(phenylsulfonyl)-2-azetidinones with N halosuccinimides. Kinetic vs. thermodynamic control. PMID- 11031052 TI - Amidine-promoted addition of chloroform to carbonyl compounds PMID- 11031053 TI - Is there a deuterium kinetic isotope effect in the one-electron transfer from 1 benzyl-1,4-dihydronicotinamide to 9-fluorenylidenemalononitrile? PMID- 11031051 TI - New building blocks for tackling the synthesis of polyhydroxylated piperidines: expeditious synthesis of amino derivatives in the 1-deoxynojirimycin series. PMID- 11031054 TI - Heterogeneous binuclear complexation of 1,3-alternate Calix PMID- 11031055 TI - Stereoselective preparation of E vinyl zirconium derivatives from E or Z enol ethers PMID- 11031056 TI - First total synthesis of grahamimycin A. PMID- 11031057 TI - Synthesis of unsymmetrical chiral triaza-18-crown-6 and diaza-12-crown-4 with a pendant group PMID- 11031058 TI - Mild and highly efficient method for the silylation of alcohols using hexamethyldisilazane catalyzed by iodine under nearly neutral reaction conditions PMID- 11031059 TI - Application of ring closing metathesis to the first total synthesis of (R)-(+) muscopyridine: determination of absolute stereochemistry PMID- 11031060 TI - Highly regioselective palladium-catalyzed beta-arylation of N, N dialkylallylamines PMID- 11031061 TI - Bicyclic lactams as chiral homoenolate equivalents: synthesis of (-)-penienone. PMID- 11031063 TI - Use of In situ generated ketene in the wynberg beta-lactone synthesis: new transformations of the dichlorinated beta-lactone products PMID- 11031062 TI - Synthesis of 4-substituted 2-phenylaminothiazoles from amidines. A convenient route to 4-trichloromethylthiazoles. PMID- 11031064 TI - Azatriquinanes. 2.(1) synthesis of azatriquinadiene and azatriquinacene PMID- 11031065 TI - Catalytic asymmetric oxidation of sulfides to sulfoxides mediated by chiral 3 substituted-1,2-benzisothiazole 1,1-dioxides PMID- 11031066 TI - Complications of parenteral nutrition at the end of the century. PMID- 11031067 TI - Nourishing the obese patient. AB - Obese patients are frequently encountered in hospitals. This is not unexpected given the fact that obesity currently constitutes a worldwide public health epidemic. The clinical indications for nutritional support and route of nutrition support selected should be similar in obese and normal weight patients. Determining nutritional requirements by conventional methods and formulas are often inaccurate in this population. For this reason, direct measurement of energy needs using indirect calorimetry when available has become the preferred method. A strategy employing hypocaloric nutrition support will be presented and discussed. PMID- 11031068 TI - Effects of dietary fish oil on survival rate, plasma amino acid pattern, and inflammatory-related mediators in diabetic rats with sepsis. AB - This study was designed to investigate the effects of dietary fish oil on survival rates, plasma amino acid profiles, and inflammatory-related mediators in diabetic rats with sepsis. Diabetes mellitus (DM) was induced in rats by streptozotocin. The DM rats were maintained for 4 weeks on medium fat (10%, w/w) diets containing either fish oil or safflower oil. After that, sepsis was induced by cecal ligation and puncture (CLP). There were 2 groups in this study: fish oil sepsis group (FOS) and safflower oil sepsis group (SOS). The survival rate was observed after CLP. Also, changes of the amino acid pattern as well as interleukin (IL)-1 beta, tumor necrosis factor (TNF)-alpha, prostaglandin (PG) E(2)at 6, 12, and 24 h after CLP were investigated. The results demonstrated that survival rates were not significantly different between the 2 groups. Plasma arginine levels were significantly lower in sepsis groups than that in the DM chow group, regardless of whether the diabetic rats were fed fish oil or safflower oil. No significant differences were observed in plasma valine, leucine, isoleucine, glutamine, or arginine concentrations between the FOS and SOS groups at different time points. Concentrations of IL-1 beta in peritoneal lavage fluid (PLF) at 6 h and TNF-alpha at 6 h as well as at 12 h after CLP in the FOS group were significantly higher than those in the SOS group. PGE(2)levels in PLF, by contrast, were lower in the FOS group at 6 and 12 h after CLP than in the SOS group. These results suggest that differences in IL-1 beta, TNF-alpha, and PGE(2)levels in PLF in the early period of sepsis did not influence the survival rates and plasma amino acid profiles of the FOS and SOS groups. Compared with safflower oil, feeding diabetic rats with fish oil had no beneficial effects on survival rates and muscle protein breakdown. The immunologic impact of dietary n-3 polyunsaturated fatty acids on diabetic rats with sepsis requires further investigation. PMID- 11031069 TI - Neither glutamine nor arginine supplementation of diets increase glutamine body stores in healthy growing rats. AB - The aim of the work was to resolve whether glutamine and arginine supplemented diets affect plasma and tissue (muscle, liver and intestinal mucosa) glutamine concentrations, as well as glutaminase and glutamine synthetase specific activities. The trial was performed in growing rats fed 10% protein diets for 3 weeks. Protein sources were: whey proteins (W); whey proteins+free glutamine (WG); whey proteins+arginine (WA); and casein+wheat protein hydrolysate+acid whey (39:39:22), as source containing protein-bound glutamine (CGW). Rats fed the control diet (6.4% glutamine) (W) showed comparable glutamine body stores to those of rats fed the WG diet. In fact, glutamine sup- plementation down regulated the hepatic glutamine synthetic capacity of growing rats (W/WG: 6.8+/ 0.3 vs 6.0+/-0.2 nmol/min/mg protein). Arginine supplementation of the diet (up to 9% of the protein content) resulted in a decrease in plasma and tissue glutamine concentrations (W/WA: plasma, 1218+/-51 vs 1031+/-48 micromol/L; liver 7.5+/-0.4 vs 6.5+/-0.2 micromol/g; muscle: 5.7+/-0.2 vs 4.0+/-0.2 micromol/g). These data suggest that glutamine supplementation of the diet does not increase plasma and tissue glutamine concentrations in healthy growing rats, while the addition of arginine to the diet decreases glutamine body stores. PMID- 11031070 TI - Effects of structurally different lipid emulsions on human neutrophil migration. AB - AIM: To test the hypothesis that structurally different lipid emulsions have distinct immunomodulatory properties, we analysed neutrophil migration in the presence of various lipid emulsions. METHOD: Neutrophils of 8 volunteers were pre incubated in medium or physiological 2.5 mM emulsions containing long-chain (LCT), medium-chain (MCT), mixed LCT/MCT, alpha -tocopherol-enriched LCT/MCT (LCT/MCT-E) or structured triglycerides (SL). Thereafter, the cells were put on top of 3 microm-pore-sized cell culture filters and incubated for one hour in the presence or absence of a chemo-attractant. Neutrophil migration was measured as the percentage of cells that had passed the filter in the presence (chemotaxis) or absence (random migration) of a chemotactic factor. RESULTS: Compared to lipid free incubation (19+/-1%) random neutrophil migration significantly decreased with LCT/MCT (11+/-2%), LCT/MCT-E (12+/-2) and MCT (5+/-2%), while LCT (18+/-3%) and SL (20+/-1%) had no effect. N-formyl-methionyl-leucyl-phenylalanine- (fMLP, 10(-8)M) or zymosan-activated-serum-induced (ZAS, 10%) filter passage under lipid free conditions amounted to 61+/-14% and 70+/-13%, respectively. These values decreased with LCT/MCT to 11+/-9% and 15+/-7%; with LCT/MCT-E to 18+/-10% and 28+/-12%; with SL to 39+/-18% and 57+/-14%, and with MCT to 5+/-2% and 10+/-6%, (all P<0.01), while LCT had no effect. Compared to LCT/MCT, the alpha -tocopherol enriched formulation significantly increased ZAS- and fMLP-induced chemotaxis. fMLP-induced chemotaxis decreased in direct proportion to LCT/MCT triglyceride concentration. CONCLUSIONS: Human neutrophil migration is distinctively inhibited by structurally different lipid emulsions, depending on triglyceride chain-length and concentration as well as alpha -tocopherol content. PMID- 11031071 TI - Serum leptin concentrations in patients with short-bowel syndrome. AB - BACKGROUND: Short-bowel syndrome is a state of severe malabsorption resulting from absence or removal of the small bowel for several causes. A number of short bowel patients develop hyperphagia. Leptin, a protein secreted from adipose tissue, signals the amount of energy stores to the brain. OBJECTIVE: To study body composition and leptin regulation in short-bowel patients and to determine whether or not leptin concentrations are linked with hyperphagia. DESIGN: We studied 25 short-bowel patients (remnant bowel less than 150 cm) and 31 controls and 10 oral nutrition. Fifteen patients received total parenteral nutrition and 10 oral nutrition. Anthropometric measurements, body composition (by bioelectrical impedance), and cholesterol, triacylglycerol and leptin concentrations were studied in all subjects. RESULTS: There were no differences between short-bowel patients and controls in anthropometric variables, body composition, or leptin concentrations. Leptin concentrations were higher in short bowel women than men (9.21+/-8.54 vs. 3.22+/-1.86 ng/ml, P=0.01). Leptin concentrations correlated positively with age (r=0.4, P=0.045), body mass index (r=0.52, P=0.007), fat mass (r=0.67, P=0.001) and body fat (r=0.68, P=0.0001); there were no correlations with other body composition parameters. We found no correlations between parenteral or oral nutrition and body composition parameters, or between leptin concentrations and the presence of hyperphagia. Logistic regression analysis showed that body fat correctly identified leptin concentrations in 60% of patients. CONCLUSIONS: Body composition, leptin concentrations and leptin regulation in patients with short-bowel syndrome are similar to those of controls. Leptin concentrations do not correlate with hyperphagia in short bowel-patients. PMID- 11031072 TI - The photodegradation of vitamins A and E in parenteral nutrition mixtures during infusion. AB - BACKGROUND & AIMS: Vitamins A and E are the most light-sensitive vitamins. Vitamin A is degraded by photolysis, while vitamin E degrades by photo-oxidation. The composition of the parenteral nutrition mixture and the container could therefore influence degradation during daylight administration. The aim of this study was therefore to determine the influence of fat emulsion and the type of bag on the photo-degradation of vitamins A and E in Parenteral Nutrition (PN) mixtures during simulated infusion in daylight. METHODS: Representative adult PN mixtures, with and without fat emulsion, were prepared. Samples for analysis were taken from infusates and each bag during simulated infusion. Degradation of vitamins A and E was determined by stability-indicating HPLC analysis. RESULTS: Results indicated that vitamin A loss proceeded rapidly during infusion, resulting in up to 80% loss in 6 hours, even with light protection of the bag. The presence of fat emulsion did not provide significant light protection. Vitamin E degradation was substantial if mixtures were prepared in EVA bags but was largely prevented if PN mixtures were compounded and stored in multi-layered bags. CONCLUSIONS: It is recommended that all PN bags should be light-protected during infusion in daylight. The use of multi-layered bags will prevent vitamin E losses during infusion. PMID- 11031073 TI - Factors associated to hypermanganesemia in patients receiving home parenteral nutrition. AB - BACKGROUND: Home parenteral nutrition (HPN) patients often present hypermanganesamia. AIM: To examine which factors may be associated to hypermanganesemia in HPN patients. METHODS: Plasma manganese (Mn), liver function tests, C-reactive protein concentrations, erythrocyte sedimentation rate (ESR), tumor necrosis factor-alpha (TNF- alpha), interleukin-6, soluble receptors of interleukin-2, and blood neopterin concentrations were determined in 21 HPN patients and 10 healthy controls. Brain magnetic resonance imaging (MRI) and careful neurologic clinical examination were performed in 11 patients. RESULTS: Mn concentration was higher in HPN patients than controls (1.96+/-1.1 vs 0.81+/- 0.4 microg/L;P<0.001) and positively correlated to the amount of parenteral nutrition (PN) supply, transaminases and alkaline phosphatase (r=0.53, P<0.0001) concentrations, as well as to ESR (r=0.61, P<0.0001), TNF- alpha and blood neopterin. The amount of calories provided by PN was positively correlated to inflammatory markers and liver parameters. All patients investigated by MRI showed hyperintense basal ganglia on T1-weighted images suggesting brain Mn deposition. Only one had slight clinical extrapyramidal symptoms. CONCLUSION: In HPN patients, sustained inflammation may facilitate hypermanganesemia through 1. cholestatic liver disease and thereby decreased Mn biliary excretion, 2. high nutritional requirements (responsible for increased Mn supply), and/or 3. modified Mn metabolism or body distribution. Neurologic complications appeared marginal whereas Mn brain deposition seems frequent. PMID- 11031074 TI - Serum levels of insulin-like growth factor-1 in the three months following surgery for a hip fracture in elderly: relationship with nutritional status and inflammatory reaction. AB - BACKGROUND AND AIMS: Serum IGF-1 level declines with advancing age and is a reliable index of protein-energy undernutrition in elderly patients in a metabolically stable condition. We have examined the value of IGF-1 as an indicator of malnutrition in elderly patients during recovery within 1 month after surgery for a hip fracture and we have studied the relationship between changes in IGF-1 levels, nutritional status and inflammatory reaction during 2 month follow up. STUDY DESIGN: Forty elderly patients (mean age 84.0+/-1.9) were nutritionally assessed upon admission to our rehabilitation unit (D0) then monthly (D30-D60) by measurement of anthropometric and biological parameters. Two groups of patients were defined on the basis of mid-arm circumference (MAC) and tricipital skinfold thickness (TST) measurement at D0: group 1, undernourished (n=13) and group 2, normally nourished (n=27). Changes in anthropometric and biological parameters during the study were compared between these two groups of patients. RESULTS: IGF-1 level was significantly lower in group 1 (79.5+/-9.1 vs 108+/-8.6 ng/ml, P<0.05) while usual serum protein levels were in the same range in the two groups. Orosomucoid level significantly decreased during 2 month follow-up in group 2 (P<0.01), CRP level decreased but not significantly. TST and MAC decreased in group 2 (respectively P<0.02 and P<0.05) while anthropometric parameters tended to slightly increase in group 1 in relation with an increase in caloric intake (P<0.05). IGF-1 level increased significantly during the study in group 2 (P<0.05) and was significantly lower in patients with complications occurring between D0 and D30 in comparison with patients without complications (respectively P<0.05 at D0, P<0.02 at D30). IGF-1 level correlated with body mass index, MAC, TST, muscle midarm circumference, albumin and transthyretin levels. Changes in IGF-1 levels positively correlated with those in transthyretin levels and negatively with those in orosomucoid levels. CONCLUSION: IGF-1 level seems a reliable index of protein-energy under-nutrition in elderly patients in the recovery period after surgery for a hip fracture. This marker is influenced by the inflammatory reaction. Decline of inflammatory reaction, rather than change in nutritional status, is responsible for the variation in IGF-1 level during a 2 month follow-up. PMID- 11031075 TI - Central venous catheter-related infections in children on long-term home parenteral nutrition: incidence and risk factors. AB - BACKGROUND AND AIMS: This study aimed to assess the incidence and etiology of central venous catheter (CVC) infections in children on home parenteral nutrition (HPN). METHODS: 207 CVC-years were studied retrospectively in 47 children on HPN, aged 8.1+/-5.0 years. RESULTS: 125 CVC were used (means: 2.6 CVC/patient and 21 months utilization/CVC). Half of the hospitalizations (162) were due to proven CVC-related infections. The mean infection incidence was 2. 1/1000 HPN days. The total population divided in two groups below and above this value: group one including 24 children, incidence < or = 2.1 per 1000 days (mean: 0.83) and group two including 23 children, incidence >2.1 per 1000 days (mean: 4.3). No differences were found between the two groups in terms of underlying disease, presence of ostomies, age at the time of HPN onset, or micro-organisms responsible. The only differences (p<0.05) were the mean duration of HPN (longer in group one) and the delay between HPN onset and the first infection (longer in group one). CONCLUSIONS: This study does not highlight any risk factors for CVC infection. However, early CVC infections after HPN onset appear to predict a bad prognosis. PMID- 11031076 TI - Bioelectrical impedance analysis during acute changes of extracellular osmolality in man. AB - BACKGROUND & AIMS: Bioelectrical impedance analysis (BIA) is widely used as an inexpensive and noninvasive method to provide estimates of body compartments such as total body water, lean body mass and fat mass. The present study was performed to test the reliability of this method during acute changes of extracellular osmolality in eight young health men. METHODS: Hyperosmolal isohydration was achieved by overnight infusions of hypertonic saline solutions (2 and 5% NaCl) and thirsting, and hypoosmolal hyperhydration by drinking of free water and overnight application of desmopressin. The control study (isoosmolality) consisted of oral water ad libitum. RESULTS: When plasma osmolality and sodium concentrations increased (from 285 +/- 1 to 296 +/- 1 mmol/kg (P<0.001) and from 141.9 +/- 0.7 to 148.3 +/- 0.6 mmol/l (P<0.0001)) and total body water remained unchanged, body impedance decreased and calculated total body water increased from 42.7 +/- 2.7 to 45.6 +/- 2.3 liters (P<0.03). In contrast, during hypoosmolal hyperhydration total body water increased by 1.56 +/- 0.17 kg and plasma osmolality decreased from 285 +/- 1 to 272 +/- 1 mmol/kg (P<0.001) and plasma sodium concentrations from 142 +/- 0.5 to 134.8 +/- 0.4 mmol/l (P<0.0001). In spite of these changes of body water, impedance measurements and calculated total body water remained unchanged. During conditions of isoosmolal isohydration (as demonstrated by unchanged plasma sodium concentrations and osmolality) the measurements by BIA also remained unchanged. CONCLUSIONS: Measurements of total body water using BIA under conditions of unknown hydration status (hyper-, hypo- or isohydration) and unknown osmolality (hyper-, hypo- or isoosmolality) may not be reliable. Therefore bioelectrical impedance analysis is not a suitable bedside method to assess changes of body compartments under unstable hydration status. PMID- 11031077 TI - Effect of cerivastatin on serum cholesterol levels in patients with type 2 diabetes mellitus. AB - The incidence of coronary heart disease (CHD) is greatly increased in overweight diabetic patients. Modification of dietary intake and weight loss improve hypercholesterolaemia. However, cholesterol goal levels are not achieved in several patients under this treatment. The aim of our study was to evaluate the effect of Cerivastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, in patients with type 2 diabetes mellitus. A population of 40 diabetic type 2 outpatients were analyzed in a prospective way. The mean+/-SD age was 60.7+/-11.6 years, with a diabetes duration of 8.5+/-6.6 years. All patients were treated with cerivastatin (0.2 mg once a day) for 6 months. Weight HbAlc fasting blood glucose, urine microalbuminuria, total cholesterol, LDL-cholesterol, HDL cholesterol and triglycerides were measured at the beginning of the study and again after 3 and 6 months of treatment with cerivastatin. An improvement in lipid levels was achieved, with a significant decrease in LDL-cholesterol (27.7%), total cholesterol (21.4%), triglycerides levels (10.4%) and a significant increase in HDL-cholesterol levels (8.3%) (P<0.05). Cardiovascular risk ratios such as; total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL cholesterol improved during treatment, decreasing 11.3% and 30%, respectively (P<0.05). Low incidence of side effects was demonstrated. In summary, cerivastatin improved lipid control in patients with type 2 diabetes, with a low incidence of side effects. PMID- 11031078 TI - Wernicke's syndrome after bariatric surgery. AB - Two young females with severe morbid obesity presented with Wernicke's syndrome after Roux-en-Y gastro-jejunum bypass had been performed. The first patient had recurrent vomiting and dyplopia two months post-surgery. Physical examination indicated bilateral ophthalmoparesia with conserved convergence and ataxia. The second patient had frequent vomiting episodes over the previous three months together with lower limb hypotonia, myoclonia and generalised tonicoclonic seizures on two occasions within one year of surgery. In both cases routine blood test, ion levels (sodium, potassium, calcium, phosphates), electroencephalogram and CT scan were normal. Thiamine therapy was instigated on the basis of clinical intuition and the first patient achieved complete remission within 24 hours while the second improved gradually in that two years later only mild lower limb hypotonia and a slight cognitive deficit remains. Erythrocyte transketolase activity determinations were abnormal on two separate occasions for this second patient. Vitamin B1 determinations were not available for the first patient. In conclusion, the restriction in energy intake and the persistent vomiting together with malabsorption induced by the surgical intervention could explain the vitamin deficiency causing Wernicke's encephalopathy. This indicates a need for close monitoring and systematic vitamin supplementation in those patients who undergo bariatric surgery. PMID- 11031079 TI - Delivery of a GDNF gene into the substantia nigra after a progressive 6-OHDA lesion maintains functional nigrostriatal connections. AB - The effects of delivering GDNF via an adenoviral vector (AdGDNF) 1 week after lesioning dopaminergic neurons in the rat substantia nigra (SN) with 6 hydroxydopamine (6-OHDA) were examined. Rats were unilaterally lesioned by injection of 6-OHDA into the striatum, resulting in progressive degeneration of dopaminergic neurons in the SN. One week later, when substantial damage had already occurred, AdGDNF or a control vector harboring beta-galactosidase (AdLacZ) was injected into either the striatum or SN (3.2 x 10(7) PFU/microl in 2 microl). Rats were examined behaviorally with the amphetamine-induced rotation test and for forelimb use for weight-bearing movements. On day 30 postlesion, the extent of nigrostriatal tract degeneration was determined by injecting a retrograde tracer (FluoroGold) bilaterally into the lesioned striatum. Five days later, rats were sacrificed within 2 h of amphetamine injection to examine amphetamine-induced Fos expression in the striatum, a measure of dopaminergic dependent function in target neurons. AdGDNF injection in the SN rescued dopaminergic neurons in the SN and increased the number of dopaminergic neurons that maintained a connection to the striatum, compared to rats injected with AdLacZ. Further support that these spared SN cells maintained functional connections to the striatum was evidenced by increased Fos expression in striatal target neurons and a decrease in amphetamine-induced rotation. In contrast to the effects observed in rats injected with AdGDNF in the SN, rats injected with AdGDNF in the striatum did not exhibit significant ameliorative effects. This study demonstrates that experimentally increasing levels of GDNF biosynthesis near the dopaminergic neuronal soma is effective in protecting the survival of these neurons and their function even when therapy is begun after 6-OHDA-induced degeneration has commenced. Thus, GDNF gene therapy may ameliorate the consequences of Parkinson's disease through rescuing compromised dopaminergic neurons. PMID- 11031080 TI - Raphe-spinal neurons display an age-dependent differential capacity for neurite outgrowth compared to other brainstem-spinal populations. AB - Functional regeneration of brainstem-spinal pathways occurs in the developing chick when the spinal cord is severed prior to embryonic day (E) 13. Functional spinal cord regeneration is not observed in animals injured after E13. This developmental transition from a permissive to a restrictive repair period may be due to the formation of an extrinsic inhibitory environment preventing axonal growth, and/or an intrinsic inability of mature neurons to regenerate. Here, we investigated the capacity of specific populations of brainstem-spinal projection neurons to regrow neurites in vitro from young (E8) versus mature (E17) brainstem explants. A crystal of carbocyanine dye (DiI) was implanted in ovo into the E5 cervical spinal cord to retrogradely label brainstem-spinal projection neurons. Three or 12 days later, discrete regions of the brainstem containing DiI-labeled neurons were dissected to produce explant cultures grown in serum-free media on laminin substrates. The subsequent redistribution of DiI into regenerating processes permitted the study of in vitro neurite outgrowth from identified brainstem-spinal neurons. When explanted on E8, i.e., an age when brainstem spinal neurons are normally elongating through the spinal cord and are capable of in vivo functional regeneration, robust neurite outgrowth was observed from all brainstem populations, including rubro-, reticulo-, vestibulo-, and raphe-spinal neurons. In contrast, when explanted on E17, robust neurite outgrowth was seen only from raphe-spinal neurons. Neurite outgrowth from raphe-spinal neurons was 5 hydroxy-tryptamine immunoreactive. This study demonstrates that in growth factor free environments with permissive growth substrates, neurite outgrowth from brainstem-spinal neurons is dependent on both neuronal age and phenotype. PMID- 11031081 TI - Increased caspase 3 and Bax immunoreactivity accompany nuclear GAPDH translocation and neuronal apoptosis in Parkinson's disease. AB - In situ end labeling combined with YOYO staining was used to mark apoptotic DNA fragmentation and chromatin condensation respectively in human postmortem brain sections. Increased numbers of apoptotic neuronal nuclei were identified in the Parkinson's disease (PD) nigra compared with age-matched controls. Caspase 3 and Bax showed increased immunoreactivity in melanized neurons of the PD nigra compared with controls. Importantly, GAPDH nuclear accumulation was also observed in the PD nigra, suggesting apoptotic rather than necrotic cell death. Interestingly, both Lewy bodies and the intranuclear Marinesco's bodies were GAPDH immunoreactive in the PD brain. PMID- 11031082 TI - Development of astroglial elements in the suprachiasmatic nucleus of the rat: with special reference to the involvement of the optic nerve. AB - The development of astroglial cells and the effect of the retinohypothalamic tract on it were studied by vimentin and glial fibrillary acidic protein (GFAP) immunocytochemistry in the suprachiasmatic nucleus (SCN) of the rat. At the embryonic stage, vimentin-immunoreactive (VIM-IR) radial glia, precursors of astrocytes, were dominant. However, their filaments vanished in the first few postnatal days. Instead of VIM-IR glial filaments, GFAP-immunoreactive (GFAP-IR) astrocytes appeared at E20 and grew rapidly from the P3 stage. GFAP immunoreactivity in the ventrolateral portion of the SCN (VLSCN) was measured using a computer-assisted image analyzing system. In normal rats, GFAP immunoreactivity showed a stepwise pattern with two slopes at P3-P4 and P20-P25. Bilaterally eye-enucleated rats operated on the day of birth showed lower GFAP immunoreactivity than normal rats and the GFAP immunoreactivity did not increase between P20 and P25 when GFAP-IR glial processes rapidly expand. Electron microscopic investigation at P50 (adult stage) revealed that neurons in the VLSCN had often direct apposition without astroglial processes and the frequency of this finding was significantly higher in eye-enucleated rats than in the control rats. These findings strongly suggest that the postnatal development of astroglial elements, particularly the expansion of GFAP-IR processes in the SCN, is regulated by retinohypothalamic projection. PMID- 11031083 TI - Inhibition of monocyte/macrophage migration to a spinal cord injury site by an antibody to the integrin alphaD: a potential new anti-inflammatory treatment. AB - The inflammatory response that ensues during the initial 48 to 72 h after spinal cord injury causes considerable secondary damage to neurons and glia. Infiltration of proinflammatory-activated neutrophils and monocytes/macrophages into the cord contributes to spinal cord injury-associated secondary damage. beta2 integrins play an essential role in leukocyte trafficking and activation and arbitrate cell-cell interactions during inflammation. The beta2 integrin, alphaDbeta2, is expressed on monocytes/macrophages and neutrophils and binds to vascular adhesion molecule-1 (VCAM-1). The increased expression of VCAM-1 during central nervous system (CNS) inflammation likely contributes to leukocyte extravasation into the CNS. Accordingly, blocking the interaction between alphaDbeta2 and VCAM-1 may attenuate the inflammatory response at the SCI site. We investigated whether the administration of monoclonal antibodies (mAbs) specific for the rat alphaD subunit would reduce the inflammatory response after a spinal cord transection injury in rats. At a 1 mg/kg dose two of three anti alphaD mAbs caused a significant ( approximately 65%) reduction in the number of macrophages at the injury site and one anti-alphaD mAb led to a approximately 43% reduction in the number of neutrophils at the SCI site. Thus, our results support the concept that the alphaDbeta2 integrins play an important role in the trafficking of leukocytes to a site of central nervous system inflammation. This study also offers preliminary evidence that anti-alphaD mAbs can reduce the extravasation of macrophages and, to a lesser extent, neutrophils, to the SCI site. PMID- 11031084 TI - The metabotropic glutamate system promotes neuronal survival through distinct pathways of programmed cell death. AB - Activation of the metabotropic glutamate receptor (mGluR) system can prevent free radical, nitric oxide (NO)-induced programmed cell death (PCD). To investigate the mechanisms utilized by the mGluR system to regulate the induction of PCD, we examined the course of PCD in real time in individual, living, primary hippocampal neurons. We assessed both phosphatidylserine (PS) externalization, an early event in PCD, and DNA fragmentation during NO toxicity and mGluR modulation to determine the individual contributions of PS externalization and genomic DNA fragmentation during neuronal PCD. Exposure to the NO donors (300 microM SNP or 300 microM NOC-9) induced PCD in approximately 75% of neurons over a 24-h period. The externalization of PS in neurons increased to 21 +/- 2% as early as 3 h following NO exposure and then increased to 80 +/- 2% over a 24-h period. The externalization of PS was independent of the loss of membrane integrity. Agonists for individual mGluR subgroups were equally able to prevent NO-induced neuronal death and DNA degradation, yet they possessed differential abilities to regulate PS externalization. The group I agonist DHPG (750 microM) and the group III agonist L-AP4 (750 microM) both prevented and reversed NO-induced PS externalization. In contrast, activation of group II subtypes using L-CCG-I (750 microM) did not prevent PS externalization. Employing an experimental model that independently led to the externalization of PS residues, we demonstrated that PS externalization does not immediately impact on neuronal survival. Yet, subsequent neuronal survival may ultimately depend upon preventing PS externalization to avoid neuronal tagging for phagocytosis. Since group I and III mGluR subtypes possess the unique ability to maintain genomic integrity and membrane PS asymmetry, these agents may provide superior overall protection against NO induced neuronal injury. PMID- 11031085 TI - Onion-bulb formation after a single compression injury in the macrophage scavenger receptor knockout mice. AB - Onion-bulb (OB) formation is often encountered in acquired neuropathies such as chronic inflammatory demyelinating polyradiculoneuropathy and diabetic neuropathy and is believed to require repeated injuries to peripheral nerves. Although this suggests that remaining damaged cell membranes, including myelin debris, might trigger OB formation, the molecular mechanism remains unclear. In this study, we were successful in producing many small OBs after a single compression injury to peripheral nerves of the knockout mice deficient of macrophage scavenger receptor class A (MSR-A). Although morphometry showed no difference in the average densities of the remaining myelinating fibers between wild-type and MSR-A knockout mice after the compression injury, there were more macrophages and myelin debris positive for oxidized-phosphatidylcholine in the nerves from the MSR-A knockout mice. We believe that OB formation was induced after a single compression injury as the result of delayed phagocytosis of myelin debris possessing oxidized lipids by MSR-A deficient macrophages. The present work shed light on the molecular mechanism of OB formation seen in chronic neuropathies and provided a model for further investigation. PMID- 11031086 TI - Attempted gene therapy for intractable pain: dexamethasone-mediated exogenous control of beta-endorphin secretion in genetically modified cells and intrathecal transplantation. AB - For optimal neural transplantation using gene engineering, it might be important to control the expression of the transfected gene extrinsically as required. This strategy could be very useful for the treatment of intractable pain that responds to opioids. For this purpose, we established a genetically modified embryonal carcinoma cell line (P19) in which the expression of beta-endorphin (beta-EP) could be controlled by the addition of dexamethasone. To obtain extrinsic control, we transfected the cells with pMAMneo containing mouse MMTV-LTR as a promoter and cDNA of the artificial beta-EP. The upregulation of beta-EP, through the activation of MMTV by the administration of dexamethasone, was confirmed in vitro. Then we transplanted these cells into the subarachonoid space in rats and evaluated the analgesic potential of these cells in vivo by hot plate test and formalin test. In the rats that received beta-EP-producing cells, we observed prominent analgesic effects after the transplantation for a month. The administration of naloxone blocked these effects. Intraperitoneal injection of 100 mg/kg dexamethasone further enhanced these effects by up to two times. These data indicate obvious analgesic effects of the cells after the transplantation and the possible exogenous upregulation of transfected beta-EP gene expression in vivo. The application of this technique might provide a new therapeutic approach to various neurological diseases. PMID- 11031087 TI - BDNF protects against spatial memory deficits following neonatal hypoxia ischemia. AB - Hypoxic-ischemic (H-I) brain injury in the human perinatal period often leads to significant long-term neurobehavioral dysfunction in the cognitive and sensory motor domains. Using a neonatal H-I injury model (unilateral carotid ligation followed by hypoxia) in postnatal day seven rats, previous studies have shown that neurotrophins, such as brain-derived neurotrophic factor (BDNF), can be protective against neural tissue loss. The present study explored potential relationships between neural protective and behavioral protective strategies in this neonatal H-I model by determining if neonatal H-I was associated with behavioral spatial learning and memory deficits and whether the neurotrophin BDNF was protective against both brain injury and spatial learning/memory dysfunction. Postnatal day seven rats received vehicle or BDNF pretreatments (intracerebroventricular injections) followed by H-I or sham treatments and then tested for spatial learning and memory on the simple place task in the Morris water maze from postnatal days 20 to 30, and their brains were histologically analyzed at 4 weeks following treatments. H-I rats with vehicle pretreatment displayed significant tissue loss in the hippocampus (including CA1 neurons), cortex, and striatum, as well as severe spatial memory deficits (e.g., short probe times). BDNF pretreatment resulted in significant protection against both H I-induced brain tissue losses and spatial memory impairments. These findings indicate that unilateral H-I brain injury in a neonatal rodent model is associated with cognitive deficits, and that BDNF pretreatment is protective against both brain injury and spatial memory impairment. PMID- 11031088 TI - Partial functional recovery of paraplegic rat by adenovirus-mediated gene delivery of constitutively active MEK1. AB - Spinal cord injury in adult mammals results in little axonal regeneration, although the mechanism of regeneration failure still remains elusive. Recent research has revealed that activation of the extracellular-signal-regulated kinases (ERKs) plays an important role in the neurite outgrowth. In the present study, we constructed a replication-defective adenovirus vector carrying mutated form of MEK1 (CA-MEK virus), which constitutively activate ERK pathway, and investigated its effect on thoracic spinal cord injury model in young adult rats as well as neurite outgrowth in vitro. In rat pheocromocytoma cell line PC12 cells, CA-MEK virus infection induced sustained activation of ERKs and stimulated neurite outgrowth in the absence of neurotrophic factors. In rat spinal cord transection model, injection of CA-MEK virus into the completely transected spinal cord efficiently activated ERKs in the supraspinal neurons and induced axonal regeneration across the transection site, which was confirmed by anterograde labeling with wheat-germ-agglutinin conjugated peroxidase (WGA-HRP). Spinal cord evoked potentials (SCEP) showed that these regenerated axons were electroconductive. Most importantly, CA-MEK virus-treated rats showed significant recovery of hind limb function 2 weeks after operation compared to the control rats treated with no virus or LacZ virus. These results suggest that adenovirus mediated CA-MEK gene transduction offers a novel strategy for the gene therapy of spinal cord injury. PMID- 11031089 TI - Reduced BDNF mRNA expression in the Parkinson's disease substantia nigra. AB - Brain-derived neurotrophic factor (BDNF) has potent effects on survival and morphology of dopaminergic neurons and thus its loss could contribute to death of these cells in Parkinson's disease (PD). In situ hybridization revealed that BDNF mRNA is strongly expressed by dopaminergic neurons in control substantia nigra pars compacta (SNpc). In clinically and neuropathologically typical PD, SNpc BDNF mRNA expression is reduced by 70% (P = 0.001). This reduction is due, in part, to loss of dopaminergic neurons which express BDNF. However, surviving dopaminergic neurons in the PD SNpc also expressed less BDNF mRNA (20%, P = 0.02) than their normal counterparts. Moreover, while 15% of control neurons had BDNF mRNA expression >1 SD below the control mean, twice as many (28%) of the surviving PD SNpc dopaminergic neurons had BDNF mRNA expression below this value. This 13% difference in proportions (95% CI 8-17%, P < or = 0.000001) indicates the presence of a subset of neurons in PD with particularly low BDNF mRNA expression. Moreover, both control and PD neurons displayed a direct relationship between the density of BDNF mRNA expression per square micrometer of cell surface and neuronal size (r(2) = 0.93, P 0.05). In contrast, apoE(-/-) mice reconstituted with apoE(-/-) bone marrow developed severe atherosclerotic lesions (453 036+/-29 767 micrometer(2), n=7) 20 weeks after transplantation. These data suggest that macrophage-derived apoE was insufficient to induce significant regression of established atherosclerotic lesions in apoE(-/-) mice, although it was sufficient to eliminate hypercholesterolemia and prevent progression of aortic lesions. PMID- 11031212 TI - Metabolic cardiovascular syndrome and risk of dementia in Japanese-American elderly men. The Honolulu-Asia aging study. AB - Cardiovascular risk factors often cluster into a metabolic syndrome that may increase the risk of dementia. The objective of the present study was to assess the long-term association between clustered metabolic cardiovascular risk factors measured at middle age and the risk of dementia in old age. This prospective cohort study of cardiovascular disease was started in 1965 and was extended to a study of dementia in 1991. The subjects were Japanese-American men with an average age of 52.7+/-4.7 (mean+/-SD) years at baseline. Dementia was diagnosed in 215 men, according to international criteria, and was based on a clinical examination, neuropsychological testing, and an informant interview. The z scores were calculated for 7 risk factors (random postload glucose, diastolic and systolic blood pressures, body mass index, subscapular skinfold thickness, random triglycerides, and total cholesterol). The relative risk (RR [95% CI]) of dementia (subtypes) per 1 SD increase in the sum of the z scores was assessed after adjustment for age, education, occupation, alcohol consumption, cigarette smoking, and years of childhood lived in Japan. The z-score sum was higher in demented subjects than in nondemented subjects, indicating a higher risk factor burden (0.74 versus -0.06, respectively; P=0. 008). Per SD increase in the z score sum, the risk of dementia was increased by 5% (RR 1.05, 95% CI 1.02 to 1.09). The z-score sum was specifically associated with vascular dementia (RR 1.11, 95% CI 1.05 to 1.18) but not with Alzheimer's disease (RR 1.00, 95% CI 0.94 to 1.05). Clustering of metabolic cardiovascular risk factors increases the risk of dementia (mainly, dementia of vascular origin). PMID- 11031214 TI - Expression of human apolipoprotein A-I/C-III/A-IV gene cluster in mice induces hyperlipidemia but reduces atherogenesis. AB - The apolipoprotein (apo)A-I/C-III/A-IV gene cluster is involved in lipid metabolism and atherosclerosis. Overexpression of apoC-III in mice causes hypertriglyceridemia and induces atherogenesis, whereas overexpression of apoA-I or apoA-IV increases cholesterol in plasma high density lipoprotein (HDL) and protects against atherosclerosis. Each gene has been studied alone in transgenic mice but not in combination as the entire cluster. To determine which phenotype is produced by the expression of the entire gene cluster, transgenic mice were generated with a 33-kb human DNA fragment. The results showed that the transgene contained the necessary elements to direct hepatic and intestinal expression of the 3 genes. In the pooled data, plasma concentrations were 257+/-9, 7.1+/-0.5, and 1.0+/-0.2 mg/dL for human apoA-I, apoC-III, and apoA-IV, respectively (mean+/ SEM). Concentrations of these apolipoproteins were higher in males than in females. Human apoA-I and apoC-III concentrations were positively correlated, suggesting that they are coregulated. Transgenic mice exhibited gross hypertriglyceridemia and accumulation of apoB(48)-containing triglyceride-rich lipoproteins. Plasma triglyceride and cholesterol concentrations were correlated positively with human apoC-III concentration, and HDL cholesterol was correlated with apoA-I concentration. In an apoE-deficient background, despite being markedly hypertriglyceridemic, cluster transgenic animals compared with nontransgenic animals showed a 61% reduction in atherosclerosis. This suggests that apoA-I and/or apoA-IV can protect against atherosclerosis even in the presence of severe hyperlipidemia. These mice provide a new model for studies of the regulation of the 3 human genes in combination. PMID- 11031215 TI - Replication of linkage of familial combined hyperlipidemia to chromosome 1q with additional heterogeneous effect of apolipoprotein A-I/C-III/A-IV locus. The NHLBI Family Heart Study. AB - Familial combined hyperlipidemia (FCHL), the most common familial dyslipidemia, is implicated in up to 20% of cases of premature coronary heart disease. Although underlying mutations for FCHL have yet to be identified, several candidate genes/regions have been identified. A positive linkage to chromosome 1q markers has been reported, with the highest lod score of 5.93 occurring at a location between D1S104 and D1S1677. Using the same diagnostic criteria, the Family Heart Study (FHS) has defined 71 FCHL families, comprising 170 cases, for a total of 137 possible affected sibling pairs. The FCHL criteria require elevation in serum low density lipoprotein cholesterol and triglyceride levels within the family, with at least 2 affected first-degree relatives. Markers D1S104 and D1S1677 were typed, and significant allele sharing was found in FCHL sibships (multipoint lod score with use of the model from the Finnish study was 2.52, and multipoint nonparametric score was 2.48; P=0.007), replicating linkage in this chromosome 1 region. In addition, previously reported linkage of FCHL to apolipoprotein A-I/C III/A-IV has been investigated in FHS families. FHS results revealed positive but nonsignificant allele sharing among FCHL sibships with apolipoprotein A-I/C-III/A IV by use of marker D11S4127 (nonparametric linkage score 1.11, P=0.13). Two locus analyses of D1S104 and D11S4127 suggested possible heterogeneity rather than epistasis, with a maximum 2-locus lod score of 3.05. A nonparametric 2-locus analysis revealed significant improvement in the 2-locus versus single-locus scores. Finally, no linkage was found with markers near the lipoprotein lipase gene region. PMID- 11031217 TI - Modulation of expression of endothelial intercellular adhesion molecule-1, platelet-endothelial cell adhesion molecule-1, and vascular cell adhesion molecule-1 in aortic arch lesions of apolipoprotein E-deficient compared with wild-type mice. AB - Human vascular adhesion molecules, such as intercellular adhesion molecule-1 (ICAM-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), and vascular cell adhesion molecule-1 (VCAM-1), are thought to play a critical role in the homing of leukocytes to sites of atherosclerotic lesions. However, very little is known about the expression of adhesion molecules in the vasculature of mice models, such as apolipoprotein E knockout (apoE(-/-)) mice, the lesions of which closely mimic human atherosclerotic lesions. This study has first quantitatively characterized the mean expression of endothelial adhesion molecules, lining the whole vessel intimal circumference, over a period of time (0 to 20 weeks of diet) in aortic arch lesions of male apoE-deficient compared with wild-type (C57BL/6) mice. These animals were fed a chow or a cholesterol-rich diet. ApoE(-/-) animals showed first an increase (at 6 weeks) and then a reduction (at 16 weeks) in the mean expression of ICAM-1 (P<0.05) and PECAM-1 (P<0.05) but not VCAM-1 levels. Such modulation of the mean expression of adhesion molecules was not observed in wild-type mice. Confirmation of immunohistochemistry results on ICAM-1 was obtained by Northern blots performed on the aortic arch of apoE and C57BL6 chow fed mice over a period of 20 weeks. Moreover, the presence of VCAM-1 was also confirmed at the RNA level, on aortas of control and apoE mice, by reverse transcription-polymerase chain reaction. In the second part of the study, we assayed the levels of adhesion molecules, in different types of histologically defined atherosclerotic lesions, in apoE(-/-) animals fed for 20 weeks. All 3 adhesion molecules (ICAM-1, PECAM-1, and VCAM-1) were observed to be reduced in fibrofatty and complex lesions but not in fatty streaks or in areas without lesions. These results indicate that the expression of these adhesion molecules in apoE-deficient animals varies with the evolution of the plaque from a fatty to a fibrous stage. PMID- 11031216 TI - Electronegative LDL from normolipemic subjects induces IL-8 and monocyte chemotactic protein secretion by human endothelial cells. AB - The presence in plasma of an electronegative LDL subfraction [LDL(-)] cytotoxic for endothelial cells (ECs) has been reported. We studied the effect of LDL(-) on the release by ECs of molecules implicated in leukocyte recruitment [interleukin 8 (IL-8) and monocyte chemotactic protein-1 (MCP-1)] and in the plasminogen activator inhibitor-1 (PAI-1). LDL(-), isolated by anion-exchange chromatography, differed from nonelectronegative LDL [LDL(+)] in its higher triglyceride, nonesterified fatty acid, apoprotein E and apoprotein C-III, and sialic acid contents. No evidence of extensive oxidation was found in LDL(-); its antioxidant and thiobarbituric acid-reactive substances contents were similar to those of LDL(+). However, conjugated dienes were increased in LDL(-), which suggests that mild oxidation might affect these particles. LDL(-) increased, in a concentration dependent manner, the release of IL-8 and MCP-1 by ECs and was a stronger inductor of both chemokines than oxidized LDL (oxLDL) or LDL(+). PAI-1 release increased slightly in ECs incubated with both LDL(-) and oxLDL but not with LDL(+). However, no cytotoxic effects of LDL(-) were observed on ECs. Actinomycin D inhibited the release of IL-8 and MCP-1 induced by LDL(-) and oxLDL by up to 80%, indicating that their production is mediated by protein synthesis. Incubation of ECs with N:-acetyl cysteine inhibited production of IL-8 and MCP-1 induced by LDL(-) and oxLDL by >50%. The free radical scavenger butylated hydroxytoluene slightly inhibited the effect of oxLDL but did not modify the effect of LDL(-). An antagonist (BN-50730) of the platelet-activating factor receptor inhibited production of both chemokines by LDL(-) and oxLDL in a concentration-dependent manner. Our results indicate that LDL(-) shows proinflammatory activity on ECs and may contribute to early atherosclerotic events. PMID- 11031218 TI - Role of the angiotensin AT(1) receptor in rat aortic and cardiac PAI-1 gene expression. AB - Although the renin-angiotensin system has been implicated in increasing plasminogen activator inhibitor-1 (PAI-1) expression, the role of the angiotensin type 1 (AT(1)) receptor is controversial. This report examines the effects of angiotensin peptides, angiotensin-converting enzyme inhibition, and AT(1) antagonism on rat aortic and cardiac PAI-1 gene expression. In vitro, angiotensin (Ang) I, Ang II, and angiotensin Arg(2)-Phe(8) (Ang III) were potent agonists of PAI-1 mRNA expression in rat aortic smooth muscle cells (RASMCs), and stimulation of PAI-1 by these peptides was blocked by the AT(1) antagonist candesartan. Angiotensin Val(3)-Phe(8) (Ang IV) and angiotensin Asp(1)-Pro(7) (Ang [1-7]) did not affect PAI-1 expression in RASMCs. In neonatal rat cardiomyocytes, Ang II increased PAI-1 mRNA expression by 4-fold (P<0.01), and this response was completely blocked by AT(1) receptor antagonism. Continuous intrajugular infusion of Ang II into Sprague-Dawley rats for 3 hours increased aortic and cardiac PAI-1 mRNA expression by 17- and 9 fold, respectively, and these Ang II responses were completely blocked by coinfusion with candesartan. Aortic and cardiac PAI-1 expressions were compared in spontaneously hypertensive rats and Wistar-Kyoto rats. PAI-1 expression in the aorta and heart from spontaneously hypertensive rats was 5.8-fold and 2-fold higher, respectively, than in control Wistar-Kyoto rats (P<0.05). Candesartan treatment for 1 week reduced aortic and cardiac PAI-1 expression in spontaneously hypertensive rats by 94% and 72%, respectively (P<0.05), but did not affect vascular PAI-1 levels in Wistar-Kyoto rats. These results demonstrate a role for the AT(1) receptor in mediating the effects of Ang II on aortic and cardiac PAI-1 gene expression. PMID- 11031219 TI - Anti-human von willebrand factor monoclonal antibody AJvW-2 prevents thrombus deposition and neointima formation after balloon injury in guinea pigs. AB - Immediately after angioplasty, platelet adhesion to the injured arterial wall and subsequent release of various mitogens may contribute to neointima formation. The purpose of this study was to evaluate the inhibitory effect of AJvW-2, a monoclonal antibody against human von Willebrand factor (vWF), on neointima formation in a guinea pig model. The carotid artery was injured with a balloon catheter, and AJvW-2 was administered by a single bolus injection. AJvW-2 dose dependently prevented neointima formation 14 days after injury. Significant inhibition was observed at 1.8 mg/kg, at which dose significant inhibition of platelet aggregation was achieved for 2 days. By elastic-Masson staining, organized thrombi were observed in the neointimal lesion on day 14. The thrombus area was significantly correlated with neointimal thickness. Furthermore, thrombus deposition, immunostained for vWF and fibrin(ogen), was observed on the media immediately after balloon injury. AJvW-2 significantly reduced the deposition of both adhesive proteins and reduced the incidence of organized thrombus formation, which might affect subsequent neointima formation. However, the proliferation of cultured smooth muscle cells was not affected by AJvW-2. These results suggest that AJvW-2 prevents neointima formation by inhibition of initial platelet-mediated thrombus formation rather than by direct inhibition of smooth muscle cell proliferation. PMID- 11031220 TI - Platelet glycoprotein IIb/IIIa receptor inhibitor preserves coronary flow reserve during progressive coronary arteriostenosis in swine. AB - Thrombosis resulting from blood platelet aggregation via glycoprotein (GP) IIb/IIIa receptor activation triggers the local release of vasoactive substances. Therefore, inhibition of these receptors could affect coronary vasoactive function during thrombotic coronary arteriostenosis. Twenty pigs were instrumented with an aortic catheter and with hydraulic occluders and flow probes on both the left anterior descending (LAD) and the left circumflex (LCx) coronary arteries. One of these 2 coronary arteries was repeatedly injured by external clamping for 15-second periods at 30-minute intervals while the pigs were given either a GP IIb/IIIa receptor inhibitor (L-739,758) (n=5), heparin (n=5), aspirin (n=3), or saline (n=7). There were no baseline differences between the 4 groups in mean arterial pressure, resting coronary blood flow (CBF), or reactive hyperemic response (RHR), which was induced by brief coronary artery occlusion and expressed as flow debt repayment. After multiple injuries, resting CBF had decreased by 95+/-2% (ie, nearly complete coronary artery occlusion) at 15+/-4 minutes in the control group, whereas in the heparin-, aspirin-, and GP IIb/IIIa inhibitor-treated groups, resting CBF had decreased by only 21+/-7% at 18+/-3 minutes, 15+/-3% at 18+/-5 minutes, and 15+/-7% at 21+/-4 minutes, respectively, suggesting that heparin, aspirin, and the GP IIb/IIIa inhibitor each prevented injury-induced coronary artery occlusion. After the initial injury, the RHR was progressively reduced in the control and heparin- and aspirin-treated groups but not in the GP IIb/IIIa inhibitor-treated group. At a comparable level of resting CBF ( approximately 15% below baseline), the RHR was reduced more in the control (-56+/-9%), heparin-treated (-49+/-9%), and aspirin-treated (-61+/-12) groups (P:<0.05) than in the GP IIb/IIIa inhibitor-treated group (-26+/-6%). When the resting CBF had decreased by approximately 35%, the RHR still was reduced significantly more (P<0.01) in the heparin-treated group (-64+/-9%) than in the GP IIb/IIIa inhibitor-treated group (-21+/-6%). In a separate group of control pigs (n=4) subjected to 2 injuries, coronary perfusion pressure distal to the injury site was reduced by 14+/-1 mm Hg from the arterial pressure, and the RHR was 20+/-6%. When the distal coronary perfusion pressure was reduced similarly ( 14+/-1 mm Hg) in a separate group of GP IIb/IIIa inhibitor-treated pigs (n=4) by 2 injuries and the use of a hydraulic occluder, the RHR was 130+/-16% (P<0.01 versus control). Our data demonstrate for the first time that a platelet GP IIb/IIIa receptor inhibitor can preserve the distal coronary vasodilatory response during progressive coronary arteriostenosis. PMID- 11031221 TI - Acute antithrombotic effect of a front-loaded regimen of clopidogrel in patients with atherosclerosis on aspirin. AB - There is a need for a rapid antithrombotic effect after the administration of antiplatelet drugs in the setting of acute coronary syndromes and percutaneous interventions. Clopidogrel, a new thienopyridine derivative, is an efficient antiplatelet agent. However, the standard regimen of clopidogrel (75 mg/d) requires 2 to 3 days before significant antithrombotic effects. Patients with stable arterial disease on chronic aspirin therapy (n=20) were treated with clopidogrel either with a front-loaded regimen, 300 mg the first day and 75 mg/d the next 7 days, or with a standard regimen, 75 mg/d for 8 days. Blood thrombogenicity was assessed by quantification of platelet-thrombus formation in an ex vivo perfusion chamber, by ADP-induced platelet aggregation, and by ADP induced fibrinogen binding. At 2 hours, mean total thrombus area with the standard regimen was not significantly reduced. In contrast, at 2 hours, the mean total thrombus area with the front-loaded regimen was significantly decreased by 23.1+/-8.5% versus baseline (P<0.05). ADP-induced platelet aggregation (with 5 and 10 micromol/L) was also significantly (P<0.05) reduced with the front-loaded regimen at 2 hours, with the mean platelet aggregation being 82.2+/-4.4% and 81.8+/-4.5%, respectively, versus baseline. Similarly, flow cytometry demonstrated a significant decrease (P<0. 05) in the ADP-induced fibrinogen binding (with 0.12 and 0.6 micromol/L) at 2 hours in this front-loaded regimen group (36.1+/-2. 0% and 53.2+/-9.3%). With the standard regimen, platelet activity was not significantly reduced at 2 hours. Our data suggest that a front loaded regimen of clopidogrel added to aspirin achieves a significant antithrombotic effect at 2 hours in patients with known atherosclerotic disease on chronic aspirin therapy. This provides a rationale for using front-loaded clopidogrel in combination with aspirin in percutaneous coronary interventions. PMID- 11031222 TI - Role of platelet P-selectin and CD40 ligand in the induction of monocytic tissue factor expression. AB - Activated platelets can express CD40 ligand (CD40L) and trigger inflammatory response and tissue factor (TF) expression in endothelial cells through interaction with CD40. This pathway is also important for T cell-induced monocyte and endothelial cell procoagulant activity. We have studied the potential role of the CD40-CD40L pathway in platelet-induced TF expression in a monocytic cell line and in whole-blood monocytes. In vitamin D(3)-differentiated U-937 cells, thrombin-stimulated platelets increased TF expression as measured by mRNA quantification, flow cytometry, and procoagulant activity. Maximum antigen expression occurred after 2 hours. Neutralizing anti-P-selectin antibody yielded a 50% suppression of procoagulant activity, whereas antibody to CD40L had no effect. In thrombin receptor activator-stimulated citrated blood, monocytes were up to 77% TF-positive, with peak expression after only 15 minutes. However, no TF mRNA was detectable at that time. Anti-P-selectin antibody reduced TF by 50%, whereas antibody to CD40L gave a 17% reduction. Thus, we conclude that P-selectin exposed on activated platelets induces the expression of TF in both U-937 cells and whole-blood monocytes but by different mechanisms. Platelet CD40L does not display any significant effect on U-937 cells but may be of some importance on whole-blood monocytes. This suggests a possible functional difference between U 937 and monocyte CD40. Another important finding in this study is the rapid appearance of surface TF on monocytes without detectable mRNA formation. This indicates that TF may be stored intracellularly in these cells and can be exposed on the surface independent of de novo protein synthesis. PMID- 11031223 TI - Oxidized LDL can promote human monocyte survival. PMID- 11031224 TI - Resistant hypertriglyceridemia in a patient with high plasma levels of apolipoprotein CII. PMID- 11031225 TI - Oxidation of apolipoprotein B-100 in circulating LDL is related to LDL residence time. In vivo insights from stable-isotope studies. AB - 5-Hydroxy-2-aminovaleric acid (HAVA) has been suggested to be a specific marker of oxidation of apolipoprotein (apo) B-100 proline (Pro) and arginine (Arg) side chain residues in low density lipoprotein (LDL) in vitro. Here we describe the application of sensitive mass spectrometric techniques to the characterization of Pro/Arg-modified apoB-100 in LDL(1) (S(f) 7 to 12) and LDL(2) (S(f) 0 to 7) in vivo. We studied 7 subjects with familial defective apoB-100 (FDB) and 8 normolipidemic controls. In FDB subjects, the presence of a mutant apoB-100 (FDB(3500Q)) in LDL markedly reduced its affinity for the LDL receptor, leading to increased residence times (RTs) of LDL(1) (65+/-21 versus 32+/-12 hours, P<0.005) and LDL(2) (230+/-40 versus 53+/-7 hours, P:<0.001) when compared with controls, as determined by stable-isotope turnover studies. LDL(1) HAVA content was not different between the groups (FDB, 0.004+/-0. 001 mol/mol apoB-100 versus controls, 0.003+/-0.001 mol/mol apoB-100, P=0.200). LDL(2) HAVA content was higher in FDB subjects (0. 374+/-0.088 versus 0.013+/-0.002 mol/mol apoB-100, P<0.001). In both groups, LDL(2) HAVA was positively associated with LDL(2) RT (FDB, r=0.893, P:=0.003; controls, r=0.976, P=0.000) and negatively correlated with LDL(2) alpha-tocopherol content (FDB, r=-0.929, P=0. 003; controls, r= 0.903, P=0.002). No significant correlations could be found between LDL(1) HAVA, LDL(1) RT, and alpha-tocopherol, respectively. The low LDL(1) HAVA content observed in both FDB and control groups was thought to be due to the relatively lower RT as well as the higher alpha-tocopherol content of these lipoproteins. In contrast, LDL(2) seemed to be strongly prone to direct oxidation of apoB-100 in vivo. The longer these particles linger in the circulation, the more apoB-100 Pro/Arg residues become modified. PMID- 11031226 TI - Effect of overexpression of human apo A-I in C57BL/6 and C57BL/6 apo E-deficient mice on 2 lipoprotein-associated enzymes, platelet-activating factor acetylhydrolase and paraoxonase. Comparison of adenovirus-mediated human apo A-I gene transfer and human apo A-I transgenesis. AB - Various mechanisms may contribute to the antiatherogenic potential of apolipoprotein A-I (apo A-I) and high density lipoproteins (HDLs). Therefore, the effect of adenovirus-mediated human apo A-I gene transfer or human apo A-I transgenesis on platelet-activating factor acetylhydrolase (PAF-AH) and arylesterase/paraoxonase (PON1) was studied in C57BL/6 and C57BL/6 apo E(-/-) mice. Human apo A-I transgenesis in C57BL/6 mice resulted in a 4.2-fold (P<0.0001) increase of PAF-AH and a 1.7-fold (P=0.0012) increase of PON1 activity. The apo E deficiency was associated with a 1.6-fold (P=0.008) lower PAF AH and a 2.0-fold (P=0.012) lower PON1 activity. Human apo A-I transgenesis in C57BL/6 apo E(-/-)mice increased PAF-AH and PON1 activity by 2.1-fold (P=0.01) and 2.5-fold (P=0.029), respectively. After adenovirus-mediated gene transfer of human apo A-I into C57BL/6 apo E(-/-)mice, a strong correlation between human apo A-I plasma levels and PAF-AH activity was observed at day 6 (r=0.92, P<0.0001). However, PON1 activity failed to increase, probably as a result of cytokine mediated inhibition of PON 1 expression. In conclusion, this study indicates that overexpression of human apo A-I increases HDL-associated PAF-AH activity. PON1 activity was also increased in human apo A-I transgenic mice, but not after human apo A-I gene transfer, a result that was probably related to cytokine production induced in the liver by the adenoviral vectors. Increased levels of these HDL associated enzymes may contribute to the anti-inflammatory and antioxidative potential of HDL and thereby to the protection conferred by HDL against atherothrombosis. PMID- 11031227 TI - R3531C mutation in the apolipoprotein B gene is not sufficient to cause hypercholesterolemia. AB - Familial hypercholesterolemia and familial ligand-defective apolipoprotein B-100 (FDB) are dominantly inherited disorders leading to impaired low-density lipoprotein receptor (LDLR) and apolipoprotein B-100 (APOB) interaction, plasma LDL elevation, and hypercholesterolemia. We previously identified the first French FDB-R3531C proband, a woman with very high total cholesterol, in a group of type IIa hypercholesterolemic families. We report here the investigation of her family at large that revealed the total absence of cosegregation with hypercholesterolemia. Six of the 10 subjects heterozygous for the R3531C mutation had plasma cholesterol lower than the 97.5th percentile for their age and gender, and mean cholesterol levels were not significantly different between affected and unaffected persons. Furthermore, 2 family members with similar high LDL cholesterol levels were not carriers of the R3531C substitution, suggesting the implication of another mutation. Segregation analysis of the LDLR gene revealed statistically significant genetic linkage with hypercholesterolemia, and analysis of the proband LDLR gene led to the identification of the 664 proline to leucine defective mutation and its detection in all 6 hypercholesterolemic-related members of this family. Therefore, our results show that the family presents with familial hypercholesterolemia and give evidence that the R3531C substitution in the APOB gene is not an allelic variant leading to FDB. Furthermore, thorough analysis of our data suggests that the APOB-R3531C mutation enhances the hypercholesterolemic effect of the LDLR-P664L defect, suggesting that it is a susceptibility mutation. PMID- 11031228 TI - Ethylene: a gaseous signal molecule in plants. AB - Ethylene regulates a multitude of plant processes, ranging from seed germination to organ senescence. Of particular economic importance is the role of ethylene as an inducer of fruit ripening. Ethylene is synthesized from S-adenosyl-L methionine via 1-aminocyclopropane-1-carboxylic acid (ACC). The enzymes catalyzing the two reactions in this pathway are ACC synthase and ACC oxidase. Environmental and endogenous signals regulate ethylene biosynthesis primarily through differential expression of ACC synthase genes. Components of the ethylene signal transduction pathway have been identified by characterization of ethylene response mutants in Arabidopsis thaliana. One class of mutations, exemplified by etr1, led to the identification of the ethylene receptors, which turned out to be related to bacterial two-component signaling systems. Mutations that eliminate ethylene binding to the receptor yield a dominant, ethylene-insensitive phenotype. CTR1 encodes a Raf-like Ser/Thr protein kinase that acts downstream from the ethylene receptor and may be part of a MAP kinase cascade. Mutants in CTR1 exhibit a constitutive ethylene-response phenotype. Both the ethylene receptors and CTR1 are negative regulators of ethylene responses. EIN2 and EIN3 are epistatic to CTR1, and mutations in either gene lead to ethylene insensitivity. Whereas the function of EIN2 in ethylene transduction is not known, EIN3 is a putative transcription factor involved in regulating expression of ethylene-responsive genes. Biotechnological modifications of ethylene synthesis and of sensitivity to ethylene are promising methods to prevent spoilage of agricultural products such as fruits, whose ripening is induced by ethylene. PMID- 11031229 TI - Mechanisms of synaptic vesicle exocytosis. AB - Chemical synaptic transmission serves as the main form of cell to cell communication in the nervous system. Neurotransmitter release occurs through the process of regulated exocytosis, in which a synaptic vesicle releases its contents in response to an increase in calcium. The use of genetic, biochemical, structural, and functional studies has led to the identification of factors important in the synaptic vesicle life cycle. Here we focus on the prominent role of SNARE (soluble NSF attachment protein receptor) proteins during membrane fusion and the regulation of SNARE function by Rab3a, nSec1, and NSF. Many of the proteins important for transmitter release have homologs involved in intracellular vesicle transport, and all forms of vesicle trafficking share common basic principles. Finally, modifications to the synaptic exocytosis pathway are very likely to underlie certain forms of synaptic plasticity and therefore contribute to learning and memory. PMID- 11031230 TI - Evolutionarily related insertion pathways of bacterial, mitochondrial, and thylakoid membrane proteins. AB - The inner membranes of eubacteria and mitochondria, as well as the chloroplast thylakoid membrane, contain essential proteins that function in oxidative phosphorylation and electron transport processes or in photosynthesis. Because most of the organellar proteins are nuclear encoded, they are synthesized in the cytoplasm and subsequently imported into the organelle before they are inserted into the membrane. This review focuses on the pathways of protein insertion into the inner membrane of eubacteria and mitochondria and into the chloroplast thylakoid membrane. In many respects, insertion of proteins into the inner membrane of bacteria is a process similar to that used by proteins of the thylakoid membrane. In both of these systems a signal recognition particle (SRP) and a SecYE-translocase are involved, as in translocation into the endoplasmic reticulum. The pathway of proteins into the mitochondrial membranes appears to be different in that it involves no SecYE-like components. A conservative pathway, recently identified in mitochondria, involves the Oxa1 protein for the insertion of proteins from the matrix. The presence of Oxa1 homologues in eubacteria and chloroplasts suggests that this pathway is evolutionarily conserved. PMID- 11031231 TI - Structural basis for the interaction of tubulin with proteins and drugs that affect microtubule dynamics. AB - The microtubule cytoskeleton is a highly regulated system. At different times in the cell cycle and positions within the organism, microtubules can be very stable or highly dynamic. Stability and dynamics are regulated by interaction with a large number of proteins that themselves may change at specific points in the cell cycle. Exogenous ligands can disrupt the normal processes by either increasing or decreasing microtubule stability and inhibiting their dynamic behavior. The recent determination of the structure of tubulin, the main component of microtubules, makes it possible now to begin to understand the details of these interactions. We review here the structure of the tubulin dimer, with particular regard to how proteins and drugs may bind and modulate microtubule dynamics. PMID- 11031232 TI - ERM-Merlin and EBP50 protein families in plasma membrane organization and function. AB - The ezrin-radixin-moesin (ERM) family of proteins have emerged as key regulatory molecules in linking F-actin to specific membrane proteins, especially in cell surface structures. Merlin, the product of the NF2 tumor suppressor gene, has sequence similarity to ERM proteins and binds to some of the same membrane proteins, but lacks a C-terminal F-actin binding site. In this review we discuss how ERM proteins and merlin are negatively regulated by an intramolecular association between their N- and C-terminal domains. Activation of at least ERM proteins can be accomplished by C-terminal phosphorylation in the presence of PIP2. We also discuss membrane proteins to which ERM and merlin bind, including those making an indirect linkage through the PDZ-containing adaptor molecules EBP50 and E3KARP. Finally, the function of these proteins in cortical structure, endocytic traffic, signal transduction, and growth control is discussed. PMID- 11031233 TI - Molecular regulation of adipogenesis. AB - Adipogenesis, or the development of fat cells from preadipocytes, has been one of the most intensely studied models of cellular differentiation. In part this has been because of the availability of in vitro models that faithfully recapitulate most of the critical aspects of fat cell formation in vivo. More recently, studies of adipogenesis have proceeded with the hope that manipulation of this process in humans might one day lead to a reduction in the burden of obesity and diabetes. This review explores some of the highlights of a large and burgeoning literature devoted to understanding adipogenesis at the molecular level. The hormonal and transcriptional control of adipogenesis is reviewed, as well as studies on a less well known type of fat cell, the brown adipocyte. Emphasis is placed, where possible, on in vivo studies with the hope that the results discussed may one day shed light on basic questions of cellular growth and differentiation in addition to possible benefits in human health. PMID- 11031234 TI - Gut feelings: enteropathogenic E. coli (EPEC) interactions with the host. AB - Enteropathogenic Escherichia coli (EPEC) is a gram-negative bacterial pathogen that adheres to human intestinal epithelial cells, resulting in watery, persistent diarrhea. It subverts the host cell cytoskeleton, causing a rearrangement of cytoskeletal components into a characteristic pedestal structure underneath adherent bacteria. In contrast to other intracellular pathogens that affect the actin cytoskeleton from inside the host cytoplasm, EPEC remains extracellular and transmits signals through the host cell plasma membrane via direct injection of virulence factors by a "molecular syringe," the bacterial type III secretion system. One injected factor is Tir, which functions as the plasma membrane receptor for EPEC adherence. Tir directly links extracellular EPEC through the epithelial membrane and firmly anchors it to the host cell actin cytoskeleton, thereby initiating pedestal formation. In addition to stimulating actin nucleation and polymerization in the host cell, EPEC activates several other signaling pathways that lead to tight junction disruption, inhibition of phagocytosis, altered ion secretion, and immune responses. This review summarizes recent developments in our understanding of EPEC pathogenesis and discusses similarities and differences between EPEC pedestals, focal contacts, and Listeria monocytogenes actin tails. PMID- 11031235 TI - Bone development. AB - Early development of the vertebrate skeleton depends on genes that pattern the distribution and proliferation of cells from cranial neural crest, sclerotomes, and lateral plate mesoderm into mesenchymal condensations at sites of future skeletal elements. Within these condensations, cells differentiate to chondrocytes or osteoblasts and form cartilages and bones under the control of various transcription factors. In most of the skeleton, organogenesis results in cartilage models of future bones; in these models cartilage is replaced by bone by the process of endochondral ossification. Lastly, through a controlled process of bone growth and remodeling the final skeleton is shaped and molded. Significant and exciting insights into all aspects of vertebrate skeletal development have been obtained through molecular and genetic studies of animal models and humans with inherited disorders of skeletal morphogenesis, organogenesis, and growth. PMID- 11031236 TI - Cellular signaling and volume control in stomatal movements in plants. AB - Stomatal guard cells are unique as a plant cell model and, because of the depth of present knowledge on ion transport and its regulation, offer a first look at signal integration in higher plants. A large body of data indicates that Ca(2+) and H(+) act independently, integrating with protein kinases and phosphatases, to control the gating of the K(+) and Cl(-) channels that mediate solute flux for stomatal movements. Oscillations in the cytosolic-free concentration of Ca(2+) contribute to a signaling cassette, integrated within these events through an unusual coupling with membrane voltage for solute homeostasis. Similar cassettes are anticipated to include control pathways linked to cytosolic pH. Additional developments during the last two years point to events in membrane traffic that play equally important roles in stomatal control. Research in these areas is now adding entirely new dimensions to our understanding of guard cell signaling. PMID- 11031237 TI - The developmental and molecular biology of genes that subdivide the body of Drosophila. AB - During the past decade, much progress has been made in understanding how the adult fly is built. Some old concepts such as those of compartments and selector genes have been revitalized. In addition, recent work suggests the existence of genes involved in the regionalization of the adult that do not have all the features of selector genes. Nevertheless, they generate morphological distinctions within the body plan. Here we re-examine some of the defining criteria of selector genes and suggest that these newly characterized genes fulfill many, but not all, of these criteria. Further, we propose that these genes can be classified according to the domains in which they function. Finally, we discuss experiments that address the molecular mechanisms by which selector and selector-like gene products function in the fly. PMID- 11031239 TI - Epithelial M cells: differentiation and function. AB - M cells are distinctive epithelial cells that occur only in the follicle associated epithelia that overlie organized mucosa-associated lymphoid tissues. They are structurally and functionally specialized for transepithelial transport, delivering foreign antigens and microorganisms to organized lymphoid tissues within the mucosae of the small and large intestines, tonsils and adenoids, and airways. M cell transport is a double-edged sword: Certain pathogens exploit the features of M cells that are intended to promote uptake for the purpose of immunological sampling. Eludication of the molecular architecture of M cell apical surfaces is important for understanding the strategies that pathogens use to exploit this pathway and for utilizing M cell transport for delivery of vaccines to the mucosal immune system. This article reviews the functional and biochemical features that distinguish M cells from other intestinal cell types. In addition it synthesizes the available information on development and differentiation of organized lymphoid tissues and the specialized epithelium associated with these immune inductive sites. PMID- 11031238 TI - Cajal bodies: the first 100 years. AB - Cajal bodies are small nuclear organelles first described nearly 100 years ago by Ramon y Cajal in vertebrate neural tissues. They have since been found in a variety of animal and plant nuclei, suggesting that they are involved in basic cellular processes. Cajal bodies contain a marker protein of unknown function, p80-coilin, and many components involved in transcription and processing of nuclear RNAs. Among these are the three eukaryotic RNA polymerases and factors required for transcribing and processing their respective nuclear transcripts: mRNA, rRNA, and pol III transcripts. A model is discussed in which Cajal bodies are the sites for preassembly of transcriptosomes, unitary particles involved in transcription and processing of RNA. A parallel is drawn to the nucleolus and the preassembly of ribosomes, which are unitary particles involved in translation of proteins. PMID- 11031240 TI - Molecular recognition and response in pollen and pistil interactions. AB - Many bisexual flowering plants possess a reproductive strategy called self incompatibility (SI) that enables the female tissue (the pistil) to reject self but accept non-self pollen for fertilization. Three different SI mechanisms are discussed, each controlled by two separate, highly polymorphic genes at the S locus. For the Solanaceae and Papaveraceae types, the genes controlling female function in SI, the S-RNase gene and the S-gene, respectively, have been identified. For the Brassicaceae type, the gene controlling male function, SCR/SP11, and the gene controlling female function, SRK, have been identified. The S-RNase based mechanism involves degradation of RNA of self-pollen tubes; the S-protein based mechanism involves a signal transduction cascade in pollen, including a transient rise in [Ca(2+)]i and subsequent protein phosphorylation/dephosphorylation; and the SRK (a receptor kinase) based mechanism involves interaction of a pollen ligand, SCR/SP11, with SRK, followed by a signal transduction cascade in the stigmatic surface cell. PMID- 11031241 TI - Retinylidene proteins: structures and functions from archaea to humans. AB - Retinylidene proteins, containing seven membrane-embedded alpha-helices that form an internal pocket in which the chromophore retinal is bound, are ubiquitous in photoreceptor cells in eyes throughout the animal kingdom. They are also present in a diverse range of other organisms and locations, such as archaeal prokaryotes, unicellular eukaryotic microbes, the dermal tissue of frogs, the pineal glands of lizards and birds, the hypothalamus of toads, and the human brain. Their functions include light-driven ion transport and phototaxis signaling in microorganisms, and retinal isomerization and various types of photosignal transduction in higher animals. The aims of this review are to examine this group of photoactive proteins as a whole, to summarize our current understanding of structure/function relationships in the best-studied examples, and to report recent new developments. PMID- 11031242 TI - Plasmodesmata: gatekeepers for cell-to-cell transport of developmental signals in plants. AB - Cell walls separate individual plant cells. To enable essential intercellular communication, plants have evolved membrane-lined channels, termed plasmodesmata, that interconnect the cytoplasm between neighboring cells. Historically, plasmodesmata were viewed as facilitating traffic of low-molecular weight growth regulators and nutrients critical to growth. Evidence for macromolecular transport via plasmodesmata was solely based on the exploitation of plasmodesmata by plant viruses during infectious spread. Now plasmodesmata are revealed to transport endogenous proteins, including transcription factors important for development. Two general types of proteins, non-targeted and plasmodesmata targeted, traffic plasmodesmata channels. Size and subcellular location influence non-targeted protein transportability. Superimposed on cargo-specific parameters, plasmodesmata themselves fluctuate in aperture between closed, open, and dilated. Furthermore, plasmodesmata alter their transport capacity temporally during development and spatially in different regions of the plant. Plasmodesmata are exposed as major gatekeepers of signaling molecules that facilitate or regulate developmental programs, maintain physiological status, and respond to pathogens. PMID- 11031243 TI - Interactions of pathogenic neisseriae with epithelial cell membranes. AB - The closely related bacterial pathogens Neisseria gonorrhoeae (gonococci, GC) and N. meningitidis (meningococci, MC) initiate infection at human mucosal epithelia. Colonization begins at apical epithelial surfaces with a multistep adhesion cascade, followed by invasion of the host cell, intracellular persistence, transcytosis, and exit. These activities are modulated by the interaction of a panoply of virulence factors with their cognate host cell receptors, and signals are sent from pathogen to host and host to pathogen at multiple stages of the adhesion cascade. Recent advances place us on the verge of understanding the colonization process at a molecular level of detail. In this review we describe the Neisseria virulence factors in the context of epithelial cell biology, placing special emphasis on the signaling functions of type IV pili, pilus-based twitching motility, and the Opa and Opc outermembrane adhesin/invasin proteins. We also summarize what is known about bacterial intracellular trafficking and growth. With the accelerated integration of tools from cell biology, biochemistry, biophysics, and genomics, experimentation in the next few years should bring unprecedented insights into the interactions of Neisseriae with their host. PMID- 11031244 TI - The role of orphan nuclear receptors in the regulation of cholesterol homeostasis. AB - Cholesterol balance is maintained by a series of regulatory pathways that control the acquisition of cholesterol from endogenous and exogenous sources and the elimination of cholesterol, facilitated by its conversion to bile acids. Over the past decade, investigators have discovered that a family of membrane-bound transcription factors, sterol regulatory element-binding proteins (SREBPs), mediate the end-product repression of key enzymes of cholesterol biosynthesis. Recently orphan members of another family of transcription factors, the nuclear hormone receptors, have been found to regulate key pathways in bile acid metabolism, thereby controlling cholesterol elimination. The study of these orphan nuclear receptors suggests their potential as targets for new drug therapies. PMID- 11031245 TI - Dynamin and its role in membrane fission. AB - Dynamin, a 100-kDa GTPase, is an essential component of vesicle formation in receptor-mediated endocytosis, synaptic vesicle recycling, caveolae internalization, and possibly vesicle trafficking in and out of the Golgi. In addition to the GTPase domain, dynamin also contains a pleckstrin homology domain (PH) implicated in membrane binding, a GTPase effector domain (GED) shown to be essential for self-assembly and stimulated GTPase activity, and a C-terminal proline-rich domain (PRD), which contains several SH3-binding sites. Dynamin partners bind to the PRD and may either stimulate dynamin's GTPase activity or target dynamin to the plasma membrane. Purified dynamin readily self-assembles into rings or spirals. This striking structural property supports the hypothesis that dynamin wraps around the necks of budding vesicles where it plays a key role in membrane fission. The focus of this review is on the relationship between the GTPase and self-assembly properties of dynamin and its cellular function. PMID- 11031246 TI - Structure and regulation of voltage-gated Ca2+ channels. AB - Voltage-gated Ca(2+) channels mediate Ca(2+) entry into cells in response to membrane depolarization. Electrophysiological studies reveal different Ca(2+) currents designated L-, N-, P-, Q-, R-, and T-type. The high-voltage-activated Ca(2+) channels that have been characterized biochemically are complexes of a pore-forming alpha1 subunit of approximately 190-250 kDa; a transmembrane, disulfide-linked complex of alpha2 and delta subunits; an intracellular beta subunit; and in some cases a transmembrane gamma subunit. Ten alpha1 subunits, four alpha2delta complexes, four beta subunits, and two gamma subunits are known. The Cav1 family of alpha1 subunits conduct L-type Ca(2+) currents, which initiate muscle contraction, endocrine secretion, and gene transcription, and are regulated primarily by second messenger-activated protein phosphorylation pathways. The Cav2 family of alpha1 subunits conduct N-type, P/Q-type, and R-type Ca(2+) currents, which initiate rapid synaptic transmission and are regulated primarily by direct interaction with G proteins and SNARE proteins and secondarily by protein phosphorylation. The Cav3 family of alpha1 subunits conduct T-type Ca(2+) currents, which are activated and inactivated more rapidly and at more negative membrane potentials than other Ca(2+) current types. The distinct structures and patterns of regulation of these three families of Ca(2+) channels provide a flexible array of Ca(2+) entry pathways in response to changes in membrane potential and a range of possibilities for regulation of Ca(2+) entry by second messenger pathways and interacting proteins. PMID- 11031247 TI - Secretory protein trafficking and organelle dynamics in living cells. AB - Green fluorescent protein chimerae acting as reporters for protein localization and trafficking within the secretory membrane system of living cells have been used in a wide variety of applications, including time-lapse imaging, double labeling, energy transfer, quantitation, and photobleaching experiments. Results from this work are clarifying the steps involved in the formation, translocation, and fusion of transport intermediates; the organization and biogenesis of organelles; and the mechanisms of protein retention, sorting, and recycling in the secretory pathway. In so doing, they are broadening our thinking about the temporal and spatial relationships among secretory organelles and the membrane trafficking pathways that operate between them. PMID- 11031248 TI - SUMO--nonclassical ubiquitin. AB - SUMO (small ubiquitin-related modifier) is the best-characterized member of a growing family of ubiquitin-related proteins. It resembles ubiquitin in its structure, its ability to be ligated to other proteins, as well as in the mechanism of ligation. However, in contrast to ubiquitination-often the first step on a one-way road to protein degradation-SUMOlation does not seem to mark proteins for degradation. In fact, SUMO may even function as an antagonist of ubiquitin in the degradation of selected proteins. While most SUMO targets are still at large, available data provide compelling evidence for a role of SUMO in the regulation of protein-protein interactions and/or subcellular localization. PMID- 11031249 TI - Viral entry into the nucleus. AB - Because many viruses replicate in the nucleus of their host cells, they must have ways of transporting their genome and other components into and out of this compartment. For the incoming virus particle, nuclear entry is often one of the final steps in a complex transport and uncoating program. Typically, it involves recognition by importins (karyopherins), transport to the nucleus, and binding to nuclear pore complexes. Although all viruses take advantage of cellular signals and factors, viruses and viral capsids vary considerably in size, structure, and in how they interact with the nuclear import machinery. Influenza and adenoviruses undergo extensive disassembly prior to genome import; herpesviruses release their genome into the nucleus without immediate capsid disassembly. Polyoma viruses, parvoviruses, and lentivirus preintegration complexes are thought to enter in intact form, whereas the corresponding complexes of onco retroviruses have to wait for mitosis because they cannot infect interphase nuclei. PMID- 11031250 TI - The Myc/Max/Mad network and the transcriptional control of cell behavior. AB - The Myc/Max/Mad network comprises a group of transcription factors whose distinct interactions result in gene-specific transcriptional activation or repression. A great deal of research indicates that the functions of the network play roles in cell proliferation, differentiation, and death. In this review we focus on the Myc and Mad protein families and attempt to relate their biological functions to their transcriptional activities and gene targets. Both Myc and Mad, as well as the more recently described Mnt and Mga proteins, form heterodimers with Max, permitting binding to specific DNA sequences. These DNA-bound heterodimers recruit coactivator or corepressor complexes that generate alterations in chromatin structure, which in turn modulate transcription. Initial identification of target genes suggests that the network regulates genes involved in the cell cycle, growth, life span, and morphology. Because Myc and Mad proteins are expressed in response to diverse signaling pathways, the network can be viewed as a functional module which acts to convert environmental signals into specific gene-regulatory programs. PMID- 11031251 TI - Cloning and recombinant expression of a structurally novel human secreted phospholipase A2. AB - Mammals contain a diverse set of secreted phospholipases A(2) (sPLA(2)s) that liberate arachidonic acid from phospholipids for the production of eicosanoids and exert a variety of physiological and pathological effects. We report the cloning, recombinant expression, and kinetic properties of a novel human sPLA(2) that defines a new structural class of sPLA(2)s called group XII. The human group XII (hGXII) cDNA contains a putative signal peptide of 22 residues followed by a mature protein of 167 amino acids that displays homology to all known sPLA(2)s only over a short stretch of amino acids in the active site region. Northern blot and reverse transcription-polymerase chain reaction analyses show that the tissue distribution of hGXII is distinct from the other human sPLA(2)s with strong expression in heart, skeletal muscle, kidney, and pancreas and weaker expression in brain, liver, small intestine, lung, placenta, ovaries, testis, and prostate. Catalytically active hGXII was produced in Escherichia coli and shown to be Ca(2+)-dependent despite the fact that it is predicted to have an unusual Ca(2+) binding loop. Similar to the previously characterized mouse group IIE sPLA(2)s, the specific activity of hGXII is low in comparison to that of other mammalian sPLA(2), suggesting that hGXII could have novel functions that are independent of its phospholipase A(2) activity. PMID- 11031252 TI - Expression of the high molecular weight fibroblast growth factor-2 isoform of 210 amino acids is associated with modulation of protein kinases C delta and epsilon and ERK activation. AB - The high molecular weight (HMW) fibroblast growth factor (FGF)-2 isoform of 210 amino acids initiated at a CUG start codon possesses a nuclear localization sequence and is not secreted. In contrast, the low molecular weight (LMW) isoform of 155 amino acids initiated at the AUG start codon can be secreted and activates the cell surface FGF receptors. The two isoforms possess different biological properties; however, little is known about the intracrine regulatory mechanisms involved in the biological effects of the HMW FGF-2 isoform. Using pancreatic cells stably transfected with cDNAs leading to the expression of either the HMW FGF-2 (A3 cells) or the LMW form (A5 cells), we provide evidence that the two FGF 2 isoforms differentially modulate PKC levels. The LMW FGF-2 up-regulated the PKC epsilon levels by 1.6-fold; by contrast the HMW isoform down-regulated the level of this PKC isotype by about 3-fold and increased the amount of PKC delta by 1.7 fold. PKC mRNAs were also modified, suggesting that PKC expression was regulated at a pretranslational level. Additionally, expression of different levels of the HMW FGF-2 with an inducible expression system confirmed the role of this isoform on PKC delta and epsilon expressions. Increased activation of ERK-1 and -2 was also observed in cells expressing the HMW FGF-2. By using different PKC inhibitors and a dominant negative PKC delta, it was found that ERK activation was PKC delta-dependent. These data indicate that expression of HMW FGF-2 can modify PKC levels by acting at the intracellular level and that the overexpression of PKC delta induces ERK-1/2 activation. The expression of a dominant negative FGFR1 did not reduce ERK-1/2 activation by the HMW FGF-2, suggesting that ERK activation does not require FGFR activity. The signaling cascade downstream of ERK might be involved in the known mitogenic effect exerted by this FGF-2 isoform. PMID- 11031253 TI - A proposed common structure of substrates bound to mitochondrial processing peptidase. AB - Mitochondrial processing peptidase (MPP), a metalloendopeptidase consisting of alpha- and beta-subunits, specifically cleaves off the N-terminal presequence of the mitochondrial protein precursor. Structural information of the substrate bound to MPP was obtained using fluorescence resonance energy transfer (FRET) measurement. A series of the peptide substrates, which have distal arginine residues required for effective cleavage at positions -7, -10, -14, and -17 from the cleavage site, were synthesized and covalently labeled with 7-diethyl aminocoumarin-3-carboxylic acid at the N termini and N,N'-dimethyl-N-(iodoacetyl) N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD) at position +4, as fluorescent donor and acceptor, respectively. When the peptides were bound to MPP, substantially the same distances were obtained between the two probes, irrespective of the length of the intervening sequence between the two probes. When 7-diethylamino-3-(4'-maleimidyl phenyl)-4-methyl coumarin was introduced into a single cysteine residue in beta-MPP as a donor and IANBD was coupled either at the N terminus or the +4 position of the peptide substrate as an acceptor, intermolecular FRET measurements also demonstrated that distances of the donor-acceptor pair were essentially the same among the peptides with different lengths of intervening sequences. The results indicate that the N terminal portion and the portion around the cleavage site of the presequence interact with specific sites in the MPP molecule, irrespective of the length of the intervening sequence between the two portions, suggesting the structure of the intervening sequence is flexible when bound to the MPP. PMID- 11031254 TI - Amyotrophic lateral sclerosis-linked glutamate transporter mutant has impaired glutamate clearance capacity. AB - We have investigated the functional impact of a naturally occurring mutation of the human glutamate transporter GLT1 (EAAT2), which had been detected in a patient with sporadic amyotrophic lateral sclerosis. The mutation involves a substitution of the putative N-linked glycosylation site asparagine 206 by a serine residue (N206S) and results in reduced glycosylation of the transporter and decreased uptake activity. Electrophysiological analysis of N206S revealed a pronounced reduction in transport rate compared with wild-type, but there was no alteration in the apparent affinities for glutamate and sodium. In addition, no change in the sensitivity for the specific transport inhibitor dihydrokainate was observed. However, the decreased rate of transport was associated with a reduction of the N206S transporter in the plasma membrane. Under ionic conditions, which favor the reverse operation mode of the transporter, N206S exhibited an increased reverse transport capacity. Furthermore, if coexpressed in the same cell, N206S manifested a dominant negative effect on the wild-type GLT1 activity, whereas it did not affect wild-type EAAC1. These findings provide evidence for a role of the N-linked glycosylation in both cellular trafficking and transport function. The resulting alteration in glutamate clearance capacity likely contributes to excitotoxicity that participates in motor neuron degeneration in amyotrophic lateral sclerosis. PMID- 11031255 TI - Dependence of soluble guanylyl cyclase activity on calcium signaling in pituitary cells. AB - The role of nitric oxide (NO) in the stimulation of soluble guanylyl cyclase (sGC) is well established, but the mechanism by which the enzyme is inactivated during the prolonged NO stimulation has not been characterized. In this paper we studied the interactions between NO and intracellular Ca(2+) in the control of sGC in rat anterior pituitary cells. Experiments were done in cultured cells, which expressed neuronal and endothelial NO synthases, and in cells with elevated NO levels induced by the expression of inducible NO synthase and by the addition of several NO donors. Basal sGC-dependent cGMP production was stimulated by the increase in NO levels in a time-dependent manner. In contrast, depolarization of cells by high K(+) and Bay K 8644, an L-type Ca(2+) channel agonist, inhibited sGC activity. Depolarization-induced down-regulation of sGC activity was also observed in cells with inhibited cGMP-dependent phosphodiesterases but not in cells bathed in Ca(2+)-deficient medium. This inhibition was independent from the pattern of Ca(2+) signaling (oscillatory versus nonoscillatory) and NO levels, and was determined by averaged concentration of intracellular Ca(2+). These results indicate that inactivation of sGC by intracellular Ca(2+) serves as a negative feedback to break the stimulatory action of NO on enzyme activity in intact pituitary cells. PMID- 11031256 TI - Mechanism of action of a novel latent membrane protein-1 dominant negative. AB - Latent membrane protein-1 (LMP1) is a signaling molecule expressed by Epstein Barr virus during latency. LMP1 is essential for B-cell immortalization by Epstein-Barr virus and transforms rodent fibroblasts. It activates many distinct signaling pathways including the transcription factors NFkappaB and AP1. We have generated a mutant of LMP1 with four point mutations; amino acids 204, 206, and 208 were mutated to alanine, and amino acid 384 was mutated to glycine. This mutant, termed LMP1(AAAG), is not only unable to activate nuclear signaling pathways, but also inhibits signaling from wild type LMP1. We have demonstrated the effectiveness, selectivity, and mechanism of this inhibitory molecule. It inhibits LMP1-stimulated NFkappaB, STAT, and Jun transcriptional activity. It is selective, as it does not inhibit TNF or interleukin-2 signaling. We have demonstrated that it does not sequester the downstream signaling molecule, TRAF2, but instead binds LMP1 and interferes with its ability to bind TRAF2. This demonstrates the importance of the interplay between the signaling domains of LMP1 and the oligomeric structure of LMP1 for effective signaling. It identifies a tool that will be useful to probe LMP1 function in disease. PMID- 11031257 TI - Blockade of the extracellular signal-regulated kinase pathway induces marked G1 cell cycle arrest and apoptosis in tumor cells in which the pathway is constitutively activated: up-regulation of p27(Kip1). AB - Constitutive activation of the ERK pathway is associated with the neoplastic phenotype of a relatively large number of human tumor cells. Blockade of the ERK pathway by treatment with PD98059, a specific inhibitor of mitogen-activated protein (MAP) kinase/ERK kinase (MEK), completely suppressed the growth of tumor cells in which the pathway is constitutively activated (RPMI-SE and HT1080 cells). Consistent with its prominent antiproliferative effect, PD98059 induced a remarkable G(1) cell cycle arrest, followed by a modest apoptotic response, in these tumor cells. Selective up-regulation of p27(Kip1) was observed after PD98059 treatment of RPMI-SE and HT1080 cells. Overexpression in RPMI-SE cells of either a kinase-negative form of MEK1 or wild-type MAP kinase phosphatase-3 also induced up-regulation of p27(Kip1). The up-regulation of p27(Kip1) correlated with increased association of p27(Kip1) with cyclin E-cyclin-dependent kinase (CDK) 2 complexes, a concomitant inhibition of cyclin E-CDK2 kinase activity, and a consequent decrease in the phosphorylation state of retinoblastoma protein, which would culminate in the marked G(1) cell cycle arrest observed in these tumor cells. These results suggest that the complete growth suppression that follows specific blockade of the ERK pathway in tumor cells in which the pathway is constitutively activated is mediated by up-regulation of p27(Kip1). PMID- 11031258 TI - SHIP1, an SH2 domain containing polyinositol-5-phosphatase, regulates migration through two critical tyrosine residues and forms a novel signaling complex with DOK1 and CRKL. AB - SHIP1 is an SH2 domain containing inositol-5-phosphatase that appears to be a negative regulator of hematopoiesis. The tyrosine kinase oncogene BCR/ABL drastically reduces expression of SHIP1. The major effect of re-expressing SHIP1 in BCR/ABL-transformed cells is reduction of hypermotility. To investigate the potential signaling pathways involving SHIP1 in hematopoietic cells, we overexpressed SHIP1 in a murine BCR/ABL-transformed Ba/F3 cell line and identified SHIP1-associated proteins. SHIP1 was found to form a novel signaling complex with BCR/ABL that includes DOK1 (p62(DOK)), phosphatidylinositol 3-kinase (PI3K), and CRKL, each of which has been previously shown to regulate migration in diverse cell types. We found that DOK1 binds directly through its PTB domain to SHIP1. Direct interaction of SHIP1 with CRKL was mediated through the CRKL-SH2 domain. Co-precipitation experiments suggest that Tyr(917) and Tyr(1020) in SHIP1 are likely to mediate interactions with DOK1. In contrast to wild type SHIP1, expression of tyrosine mutant SHIP1 by transient transfection did not alter migration. PI3K was likely linked to this complex by CRKL. Thus, this complex may serve to generate a very specific set of phosphoinositol products, possibly involved in regulating migration. Overall, these data suggest that proteins that interact with SHIP1 through Tyr(917) and Tyr(1020), such as DOK1 and SHC, are likely to be involved in the regulation of SHIP1 dependent migration. PMID- 11031259 TI - Cell autonomous apoptosis defects in acid sphingomyelinase knockout fibroblasts. AB - A body of evidence suggests that stress-induced sphingomyelin hydrolysis to the second messenger ceramide initiates apoptosis in some cells. Although studies using lymphoblasts from Niemann-Pick disease patients or acid sphingomyelinase (ASMase)-deficient mice have provided genetic support for this hypothesis, these models have not been universally accepted as definitive. Here, we show that mouse embryonic fibroblasts (MEFs) prepared from asmase mice manifest cell autonomous defects in apoptosis in response to several stresses. In particular, asmase(-/-) MEFs failed to generate ceramide and were totally resistant to radiation-induced apoptosis but remained sensitive to staurosporine, which did not induce ceramide. asmase(-/-) MEFs were also partially resistant to tumor necrosis factor alpha/ actinomycin D and serum withdrawal. Thus, resistance to apoptosis in asmase(-/-) MEFs was not global but rather stress type specific. Most importantly, the sensitivity to stress could be restored in the asmase(-/-) MEFs by administration of natural ceramide. Overcoming apoptosis resistance by natural ceramide is evidence that it is the lack of ceramide, not ASMase, that determines apoptosis sensitivity. The ability to rescue the apoptotic phenotype without reversing the genotype by the product of the enzymatic deficiency provides proof that ceramide is obligate for apoptosis induction in response to some stresses. PMID- 11031260 TI - Acylation of lysine 983 is sufficient for toxin activity of Bordetella pertussis adenylate cyclase. Substitutions of alanine 140 modulate acylation site selectivity of the toxin acyltransferase CyaC. AB - The capacity of adenylate cyclase toxin (ACT) to penetrate into target cells depends on post-translational fatty-acylation by the acyltransferase CyaC, which can palmitoylate the conserved lysines 983 and 860 of ACT. Here, the in vivo acylating capacity of a set of mutated CyaC acyltransferases was characterized by two-dimensional gel electrophoresis and mass spectrometric analyses of the ACT product. Substitutions of the potentially catalytic serine 20 and histidine 33 residues ablated acylating activity of CyaC. Conservative replacements of alanine 140 by glycine (A140G) and valine (A140V) residues, however, affected selectivity of CyaC for the two acylation sites on ACT. Activation by the A140G variant of CyaC generated a mixture of bi- and monoacylated ACT molecules, modified either at both Lys-860 and Lys-983, or only at Lys-860, respectively. In contrast, the A140V CyaC produced a nearly 1:1 mixture of nonacylated pro-ACT with ACT monoacylated almost exclusively at Lys-983. The respective proportion of toxin molecules acylated at Lys-983 correlated well with the cell-invasive activity of both ACT mixtures, which was about half of that of ACT fully acylated on Lys-983 by intact CyaC. These results show that acylation of Lys-860 alone does not confer cell-invasive activity on ACT, whereas acylation of Lys-983 is necessary and sufficient. PMID- 11031261 TI - Tigerinins: novel antimicrobial peptides from the Indian frog Rana tigerina. AB - Four broad-spectrum, 11 and 12 residue, novel antimicrobial peptides have been isolated from the adrenaline-stimulated skin secretions of the Indian frog Rana tigerina. Sequences of these peptides have been determined by automated Edman degradation, by mass spectral analysis and confirmed by chemical synthesis. These peptides, which we have named as tigerinins, are characterized by an intramolecular disulfide bridge between two cysteine residues forming a nonapeptide ring. This feature is not found in other amphibian peptides. Conformational analysis indicate that the peptides tend to form beta-turn structures. The peptides are cationic and exert their activity by permeabilizing bacterial membranes. Tigerinins represent the smallest, nonhelical, cationic antimicrobial peptides from amphibians. PMID- 11031262 TI - Recycling of the insulin-sensitive glucose transporter GLUT4. Access of surface internalized GLUT4 molecules to the perinuclear storage compartment is mediated by the Phe5-Gln6-Gln7-Ile8 motif. AB - The insulin-sensitive glucose transporter GLUT4 is translocated to the plasma membrane in response to insulin and recycled back to the intracellular store(s) after removal of the hormone. We have used clonal 3T3-L1 fibroblasts and adipocyte-like cells stably expressing wild-type GLUT4 to characterize (a) the intracellular compartment where the bulk of GLUT4 is intracellularly stored and (b) the mechanisms involved in the recycling of endocytosed GLUT4 to the store compartment. Surface internalized GLUT4 is targeted to a large, flat, fenestrated saccular structure resistant to brefeldin A that localized to the vicinity of the Golgi complex is sealed to endocytosed transferrin (GLUT4 storage compartment). Recycling of endocytosed GLUT4 was studied by comparing the cellular distributions of antibody/biotin tagged GLUT4 and GLUT4(Ser(5)), a mutant with the Phe(5)-Gln(6)-Gln(7)-Ile(8) inactivated by the substitution of Ser for Phe(5). Ablation of the Phe(5)-Gln(6)-Gln(7)-Ile(8) inhibits the recycling of endocytosed GLUT4 to the GLUT4 store compartment and results in its transport to late endosomes/lysosomes where it is rapidly degraded. PMID- 11031263 TI - The coenzyme b12 analog 5'-deoxyadenosylcobinamide-gdp supports catalysis by methylmalonyl-coa mutase in the absence of trans-ligand coordination. AB - Methylmalonyl-CoA mutase is an 5'-adenosylcobalamin (AdoCbl)-dependent enzyme that catalyzes the rearrangement of methylmalonyl-CoA to succinyl-CoA. The crystal structure of this protein revealed that binding of the cofactor is accompanied by a significant conformational change in which dimethylbenzimidazole, the lower axial ligand to cobalt in solution, is replaced by His(610) donated by the active site. The role of the lower axial ligand in the trillion-fold labilization of the upper axial cobalt-carbon bond has been the subject of enduring debate in the model inorganic literature. In this study, we have used a cofactor analog, 5'deoxyadenosylcobinamide GDP (AdoCbi-GDP), which reconstitutes the enzyme in a "histidine-off" form and which allows us to evaluate the contribution of the lower axial ligand to catalysis. The k(cat) for the enzyme in the presence of AdoCbi-GDP is reduced by a factor of 4 compared with the native cofactor AdoCbl. The overall deuterium isotope effect in the presence of AdoCbi-GDP ((D)V = 7.2 +/- 0.8) is comparable with that observed in the presence of AdoCbl (5.0 +/- 0.6) and indicates that the hydrogen transfer steps in this reaction are not significantly affected by the change in coordination state of the bound cofactor. These surprising results are in marked contrast to the effects ascribed to the corresponding lower axial histidine ligands in the cobalamin-dependent enzymes glutamate mutase and methionine synthase. PMID- 11031264 TI - African swine fever virus protease, a new viral member of the SUMO-1-specific protease family. AB - African swine fever virus (ASFV) is a complex DNA virus that employs polyprotein processing at Gly-Gly-Xaa sites as a strategy to produce several major core components of the viral particle. The virus gene S273R encodes a 31-kDa protein that contains a "core domain" with the conserved catalytic residues characteristic of SUMO-1-specific proteases and the adenovirus protease. Using a COS cell expression system, it was found that protein pS273R is capable of cleaving the viral polyproteins pp62 and pp220 in a specific way giving rise to the same intermediates and mature products as those produced in ASFV-infected cells. Furthermore, protein pS273R, like adenovirus protease and SUMO-1-specific enzymes, is a cysteine protease, because its activity is abolished by mutation of the predicted catalytic histidine and cysteine residues and is inhibited by sulfhydryl-blocking reagents. Protein pS273R is expressed late after infection and is localized in the cytoplasmic viral factories, where it is found associated with virus precursors and mature virions. In the virions, the protein is present in the core shell, a domain where the products of the viral polyproteins are also located. The identification of the ASFV protease will allow a better understanding of the role of polyprotein processing in virus assembly and may contribute to our knowledge of the emerging family of SUMO-1-specific proteases. PMID- 11031265 TI - Increased production of beta-amyloid and vulnerability to endoplasmic reticulum stress by an aberrant spliced form of presenilin 2. AB - An alternative spliced form of the presinilin 2 (PS2) gene (PS2V) lacking exon 5 has previously been reported to be expressed in human brains in sporadic Alzheimer's disease (AD). PS2V encodes the amino-terminal portion of PS2, which contains residues Met1-Leu119 and 5 additional amino acid residues (SSMAG) at its carboxyl terminus. Here we report that PS2V protein impaired the signaling pathway of the unfolded protein response, similarly to familial AD-linked PS1 mutants and caused significant increases in the production of both amyloid beta40 and beta42. Interestingly, PS2V-encoding protein was expressed in neuropathologically affected neurons of the hippocampal CA1 region and temporal cortex in AD patients. These findings suggest that the aberrant splicing of the PS2 gene may be implicated in the neuropathology of sporadic AD. PMID- 11031266 TI - Structures of prolyl oligopeptidase substrate/inhibitor complexes. Use of inhibitor binding for titration of the catalytic histidine residue. AB - Structure determination of the inactive S554A variant of prolyl oligopeptidase complexed with an octapeptide has shown that substrate binding is restricted to the P4-P2' region. In addition, it has revealed a hydrogen bond network of potential catalytic importance not detected in other serine peptidases. This involves a unique intramolecular hydrogen bond between the P1' amide and P2 carbonyl groups and another between the P2' amide and Nepsilon2 of the catalytic histidine 680 residue. It is argued that both hydrogen bonds promote proton transfer from the imidazolium ion to the leaving group. Another complex formed with the product-like inhibitor benzyloxycarbonyl-glycyl-proline, indicating that the carboxyl group of the inhibitor forms a hydrogen bond with the Nepsilon2 of His(680). Because a protonated histidine makes a stronger interaction with the carboxyl group, it offers a possibility of the determination of the real pK(a) of the catalytic histidine residue. This was found to be 6.25, lower than that of the well studied serine proteases. The new titration method gave a single pK(a) for prolyl oligopeptidase, whose reaction exhibited a complex pH dependence for k(cat)/K(m), and indicated that the observed pK(a) values are apparent. The procedure presented may be applicable for other serine peptidases. PMID- 11031267 TI - Superimposed promoter sequences of the adenoviral E2 early RNA polymerase III and RNA polymerase II transcription units. AB - The human adenovirus type 2 E2 early (E2E) transcriptional control region contains an efficient RNA polymerase III promoter, in addition to the well characterized promoter for RNA polymerase II. To determine whether this promoter includes intragenic sequences, we examined the effects of precise substitutions introduced between positions +2 and +62 on E2E transcription in an RNA polymerase III-specific, in vitro system. Two noncontiguous sequences within this region were necessary for efficient or accurate transcription by this enzyme. The sequence and properties of the functional element proximal to the sites of initiation identified it as an A box. Although a B box sequence could not be unambiguously located, substitutions between positions +42 and +62 that severely impaired transcription also inhibited binding of the human general initiation protein TFIIIC. Thus, this region of the RNA polymerase III E2E promoter contains a B box sequence. We also identified previously unrecognized intragenic sequences of the E2E RNA polymerase II promoter. In conjunction with our previous observations, these data establish that RNA polymerase II and RNA polymerase III promoter sequences are superimposed from approximately positions -30 to +20 of the complex E2E transcriptional control region. The alterations in transcription induced by certain mutations suggest that components of the RNA polymerase II and RNA polymerase III transcriptional machines compete for access to overlapping binding sites in the E2E template. PMID- 11031268 TI - Expression of a glutamate decarboxylase homologue is required for normal oxidative stress tolerance in Saccharomyces cerevisiae. AB - The action of gamma-aminobutyrate (GABA) as an intercellular signaling molecule has been intensively studied, but the role of this amino acid metabolite in intracellular metabolism is poorly understood. In this work, we identify a Saccharomyces cerevisiae homologue of the GABA-producing enzyme glutamate decarboxylase (GAD) that is required for normal oxidative stress tolerance. A high copy number plasmid bearing the glutamate decarboxylase gene (GAD1) increases resistance to two different oxidants, H(2)O(2) and diamide, in cells that contain an intact glutamate catabolic pathway. Structural similarity of the S. cerevisiae GAD to previously studied plant enzymes was demonstrated by the cross-reaction of the yeast enzyme to a antiserum directed against the plant GAD. The yeast GAD also bound to calmodulin as did the plant enzyme, suggesting a conservation of calcium regulation of this protein. Loss of either gene encoding the downstream steps in the conversion of glutamate to succinate reduced oxidative stress tolerance in normal cells and was epistatic to high copy number GAD1. The gene encoding succinate semialdehyde dehydrogenase (UGA5) was identified and found to be induced by H(2)O(2) exposure. Together, these data strongly suggest that increases in activity of the glutamate catabolic pathway can act to buffer redox changes in the cell. PMID- 11031269 TI - Probing essential water in yeast pyrophosphatase by directed mutagenesis and fluoride inhibition measurements. AB - The pattern of yeast pyrophosphatase (Y-PPase) inhibition by fluoride suggests that it replaces active site Mg(2+)-bound nucleophilic water, for which two different locations were proposed previously. To localize the bound fluoride, we investigate here the effects of mutating Tyr(93) and five dicarboxylic amino acid residues forming two metal binding sites in Y-PPase on its inhibition by fluoride and its five catalytic functions (steady-state PP(i) hydrolysis and synthesis, formation of enzyme-bound PP(i) at equilibrium, phosphate-water oxygen exchange, and Mg(2+) binding). D117E substitution had the largest effect on fluoride binding and made the P-O bond cleavage step rate-limiting in the catalytic cycle, consistent with the mechanism in which the nucleophile is coordinated by two metal ions and Asp(117). The effects of the mutations on PP(i) hydrolysis (as characterized by the catalytic constant and the net rate constant for P-O bond cleavage) were in general larger than on PP(i) synthesis (as characterized by the net rate constant for PP(i) release from active site). The effects of fluoride on the Y-PPase variants confirmed that PPase catalysis involves two enzyme.PP(i) intermediates, which bind fluoride with greatly different rates (Baykov, A. A., Fabrichniy, I. P., Pohjanjoki, P., Zyryanov, A. B., and Lahti, R. (2000) Biochemistry 39, 11939-11947). A mechanism for the structural changes underlying the interconversion of the enzyme.PP(i) intermediates is proposed. PMID- 11031270 TI - De novo activation of the beta-phaseolin promoter by phosphatase or protein synthesis inhibitors. AB - The promoter for the phaseolin (phas) bean seed protein gene adopts an inactive chromatin structure in leaves of transgenic tobacco. This repressive architecture, which confers stringent spatial regulation, is disrupted upon transcriptional activation during embryogenesis in a process that requires the presence of both a transcription factor (PvALF) and abscisic acid (ABA). Toward determining the need for de novo synthesis of proteins other than PvALF in transcriptional activation we explored the effect of several eukaryotic protein synthesis inhibitors. Surprisingly, cycloheximide (CHX), emetine, and verrucarin A were able to induce transcription from the phas promoter in tobacco and bean leaf tissue in the absence of either PvALF or ABA. This induction was decreased by the replication inhibitors hydroxyurea and aphidicolin but not by genistein or mimosine. Since protein phosphatases and kinases are essential components of the ABA signal transduction pathway, it is conceivable that CHX is also capable of inducing phosphorylation of proteins usually involved in ABA-mediated activation. Interestingly, okadaic acid, an inhibitor of serine/threonine phosphatase, also strongly activated transcription from the phas promoter. In contrast, the protein synthesis inhibitors anisomycin and puromycin did not activate transcription from the phas promoter, nor did the tyrosine phosphatase inhibitors phenylarsine oxide and sodium orthovanadate. These discrete but different results on transcriptional activation may reflect specific modes of action of the inhibitors, or they may reflect differential interactions of the inhibitors or of downstream events resulting from inhibitor activity with presently unknown components of the transcriptional activation system. PMID- 11031271 TI - Charged residues at the intracellular boundary of transmembrane helices 2 and 3 independently affect constitutive activity of Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor. AB - Because charged residues at the intracellular ends of transmembrane helix (TMH) 2 and TMH3 of G protein-coupled receptors (GPCRs) affect signaling, we performed mutational analysis of these residues in the constitutively signaling Kaposi's sarcoma-associated herpesvirus GPCR (KSHV-GPCR). KSHV-GPCR contains the amino acid sequence Val-Arg-Tyr rather than the Asp/Glu-Arg-Tyr ((D/E)RY) motif at the intracellular end of TMH3. Mutation of Arg-143 to Ala (R143A) or Gln (R143Q) abolished constitutive signaling whereas R143K exhibited 50% of the basal activity of KSHV-GPCR. R143A was not stimulated by agonist, whereas R143Q was stimulated by growth-related oncogene-alpha, and R143K, similar to KSHV-GPCR, was stimulated further. These findings show that Arg-143 is critical for signal generation in KSHV-GPCR. In other GPCRs, Arg in this position may act as a signaling switch by movement of its sidechain from a hydrophilic pocket in the TMH bundle to a position outside the bundle. In rhodopsin, the Arg of Glu-Arg-Tyr interacts with the adjacent Asp to constrain Arg outside the TMH bundle. V142D was 70% more active than KSHV-GPCR, suggesting that an Arg residue, which is constrained outside the bundle by interacting with Asp-142, leads to a receptor that signals more actively. Because the usually conserved Asp in the middle of TMH2 is not present in KSHV-GPCR, we tested whether Asp-83 at the intracellular end of TMH2 was involved in signaling. D83N and D83A were 110 and 190% more active than KSHV-GPCR, respectively. The double mutant D83A/V142D was 510% more active than KSHV-GPCR. That is, cosubstitutions of Asp-83 by Ala and Val-142 by Asp act synergistically to increase basal signaling. A model of KSHV-GPCR predicts that Arg-143 interacts with residues in the TMH bundle and that the sidechain of Asp-83 does not interact with Arg-143. These data are consistent with the hypothesis that Arg-143 and Asp-83 independently affect the signaling activity of KSHV-GPCR. PMID- 11031272 TI - Overexpression of neuronal pentraxin 1 is involved in neuronal death evoked by low K(+) in cerebellar granule cells. AB - Mature cerebellar granule cells in culture die by a process that requires new RNA and protein synthesis when deprived of depolarizing concentrations of potassium. We investigated gene expression during the early phase of the cell death program evoked by potassium deprivation. Using a differential gene display technique, we isolated a cDNA that was increased by potassium deprivation. This cDNA was homologous to the 3' mRNA end of neuronal pentraxin 1 (NP1), a gene encoding a secreted glycoprotein whose expression is restricted to the nervous system. Reverse-Northern and Northern blot analyses confirmed that treatment with low potassium induces overexpression of NP1 mRNA, with a subsequent increase in NP1 protein levels. Time-course studies indicated that overexpression of NP1 protein reaches a maximum after 4 h of exposure to potassium deprivation and 4 h before significant cell death. Incubation of cerebellar granule cells with an antisense oligodeoxyribonucleotide directed against NP1 mRNA reduced low potassium-evoked NP1 protein levels by 60% and attenuated neuronal death by 50%, whereas incubation with the corresponding sense oligodeoxyribonucleotide was ineffective. Furthermore, acute treatment with lithium significantly inhibited both overexpression of NP1 and cell death evoked by low potassium. These results indicate that NP1 is part of the gene expression program of apoptotic cell death activated by nondepolarizing culture conditions in cerebellar granule cells. PMID- 11031274 TI - Fifty years in physical chemistry: homage to mentors, methods, and molecules. AB - A nostalgic account is given of my scientific odyssey, recalling early encounters, some fateful, some just fun, with mentors, methods, and molecules. These include stories of my student years at Stanford, pursuing chemical kinetics with Harold Johnston; graduate study at Harvard, doing molecular spectroscopy with Bright Wilson; and fledgling faculty years at Berkeley, launching molecular beam studies of reaction dynamics. A few vignettes from my "ever after " era on the Harvard faculty emphasize thematic motivations or methods inviting further exploration. An Appendix provides a concise listing of colleagues in research and the topics we have pursued. PMID- 11031273 TI - Affinity labeling of rat glutathione S-transferase isozyme 1-1 by 17beta iodoacetoxy-estradiol-3-sulfate. AB - Rat liver glutathione S-transferase, isozyme 1-1, catalyzes the glutathione dependent isomerization of Delta(5)-androstene-3,17-dione and also binds steroid sulfates at a nonsubstrate inhibitory steroid site. 17beta-Iodoacetoxy-estradiol 3-sulfate, a reactive steroid analogue, produces a time-dependent inactivation of this glutathione S-transferase to a limit of 60% residual activity. The rate constant for inactivation (k(obs)) exhibits a nonlinear dependence on reagent concentration with K(I) = 71 microm and k(max) = 0.0133 min(-1). Complete protection against inactivation is provided by 17beta-estradiol-3,17-disulfate, whereas Delta5-androstene-3,17-dione and S-methylglutathione have little effect on k(obs). These results indicate that 17beta-iodoacetoxy-estradiol-3-sulfate reacts as an affinity label of the nonsubstrate steroid site rather than of the substrate sites occupied by Delta5-androstene-3,17-dione or glutathione. Loss of activity occurs concomitant with incorporation of about 1 mol 14C-labeled reagent/mol enzyme dimer when the enzyme is maximally inactivated. Isolation of the labeled peptide from the chymotryptic digest shows that Cys(17) is the only enzymic amino acid modified. Covalent modification of Cys(17) by 17beta iodoacetoxy-estradiol-3-sulfate on subunit A prevents reaction of the steroid analogue with subunit B. These results and examination of the crystal structure of the enzyme suggest that the interaction between the two subunits of glutathione S-transferase 1-1, and the electrostatic attraction between the 3 sulfate of the reagent and Arg(14) of subunit B, are important in binding steroid sulfates at the nonsubstrate steroid binding site and in determining the specificity of this affinity label. PMID- 11031275 TI - Surface plasmon resonance imaging measurements of ultrathin organic films. AB - The surface-sensitive optical technique of surface plasmon resonance (SPR) imaging is used to characterize ultrathin organic and biopolymer films at metal interfaces in a spatially resolved manner. Because of its high surface sensitivity and its ability to measure in real time the interaction of unlabeled biological molecules with arrays of surface-bound species, SPR imaging has the potential to become a powerful tool in biomolecular investigations. Recently, SPR imaging has been successfully implemented in the characterization of supported lipid bilayer films, the monitoring of antibody-antigen interactions at surfaces, and the study of DNA hybridization adsorption. The following is included in this review: (a) an introduction to the principles of surface plasmon resonance, (b) the details of SPR imaging instrumental design, (c) a short discussion concerning resolution, sensitivity, and quantitation in SPR imaging, (d) the details of DNA array fabrication on chemically modified gold surfaces, and (e) two examples that demonstrate the application of the SPR imaging technique to the study of protein DNA interactions. PMID- 11031276 TI - Delayed ionization and fragmentation en route to thermionic emission: statistics and dynamics. AB - Thermionic emission is discussed as a long time (microseconds) decay mode of energy-rich large molecules, metallic and metcar clusters, and fullerenes. We review what is known and consider the many experiments, systems, and theoretical and computational studies that still need to be done. We conclude with a wish list for future work. Particular attention is given to the experimental signatures, such as the dependence on the mode of energy acquisition, and theoretical indications of a not-quite-statistical delayed ionization and to the competition of electron emission with other decay modes, such as fragmentation or radiative cooling. Coupling of the electronic and nuclear modes can be a bottleneck and quite long time-delayed ionization can be observed, as in the decay of high Rydberg states probed by ZEKE spectroscopy, before the onset of complete energy partitioning. PMID- 11031277 TI - Spatially heterogeneous dynamics in supercooled liquids. AB - Although it has long been recognized that dynamics in supercooled liquids might be spatially heterogeneous, only in the past few years has clear evidence emerged to support this view. As a liquid is cooled far below its melting point, dynamics in some regions of the sample can be orders of magnitude faster than dynamics in other regions only a few nanometers away. In this review, the experimental work that characterizes this heterogeneity is described. In particular, the following questions are addressed: How large are the heterogeneities? How long do they last? How much do dynamics vary between the fastest and slowest regions? Why do these heterogeneities arise? The answers to these questions influence practical applications of glass-forming materials, including polymers, metallic glasses, and pharmaceuticals. PMID- 11031278 TI - Generalized born models of macromolecular solvation effects. AB - It would often be useful in computer simulations to use a simple description of solvation effects, instead of explicitly representing the individual solvent molecules. Continuum dielectric models often work well in describing the thermodynamic aspects of aqueous solvation, and approximations to such models that avoid the need to solve the Poisson equation are attractive because of their computational efficiency. Here we give an overview of one such approximation, the generalized Born model, which is simple and fast enough to be used for molecular dynamics simulations of proteins and nucleic acids. We discuss its strengths and weaknesses, both for its fidelity to the underlying continuum model and for its ability to replace explicit consideration of solvent molecules in macromolecular simulations. We focus particularly on versions of the generalized Born model that have a pair-wise analytical form, and therefore fit most naturally into conventional molecular mechanics calculations. PMID- 11031279 TI - Chemical dynamics at metal surfaces. AB - Theoretical aspects of dynamical processes at metal surfaces are reviewed. Experimental challenges to theory are presented and progress toward meeting these challenges is appraised. Topics include adsorbate vibrational energy flow, inelastic molecule-surface scattering, adsorption, transient mobility, dissociation, desorption, photochemistry, and electron-induced chemistry at metal surfaces. Experimental examples cited illustrate the richness of dynamical phenomena to be understood and the necessity of developing multidimensional, beyond Born-Oppenheimer, formulations of adsorbate dynamics. Classical mechanical and quantum mechanical treatments of dynamics are contrasted. The importance of including phonon and electron-hole pair dissipation in theories of adsorbate dynamics is emphasized, and strategies for doing this in classical and quantum treatments are presented. PMID- 11031280 TI - Peptides and proteins in the vapor phase. AB - This article provides a review of recent studies of the properties of unsolvated (and partially solvated) peptides and proteins. The methods used to produce vapor phase peptide and protein ions are described along with some of the techniques used to study them, such as H/D exchange, blackbody infrared radiative dissociation, and ion mobility measurements. Studies of unsolvated peptides and proteins provide information about their intrinsic intramolecular interactions. The topics covered include the role of zwitterions and salt bridges in the vapor phase, Coulomb interactions in multiply charged ions, the unfolding and refolding of vapor-phase proteins, and the stability of unsolvated helices and sheets. Finally, dehydration and rehydration studies of proteins in the vapor phase are described. These can provide exquisitely detailed information about hydration interactions, such as the enthalpy and entropy changes associated with adsorbing individual water molecules. PMID- 11031281 TI - Effective interactions between electric double layers. AB - This review summarizes and assesses recent theoretical and experimental advances, with special emphasis on the effective interaction between charge-stabilized colloids, in the bulk or in confined geometries, and on the ambiguities of defining an effective charge of the colloidal particles. Some consideration is given to the often neglected discrete solvent effects. PMID- 11031282 TI - Transient laser frequency modulation spectroscopy. AB - Explicitly time-dependent implementations of optical frequency modulation spectroscopy have been recently applied to a wide range of problems in chemical physics. We provide a brief description of the methodology, with an emphasis on its intrinsic advantages for interrogating transient species. Several examples highlight the application of the technique to high-resolution absorption spectra of free radicals, rate measurements for gas-phase reactions, and Doppler spectroscopy of the gas-phase products of photoinitiated reactions. PMID- 11031283 TI - Motion and disorder in crystal structure analysis: measuring and distinguishing them. AB - Dynamic processes in crystalline solids are reflected in the atomic displacement amplitudes determined, together with the atomic coordinates, by crystal structure analysis. The interpretation of such amplitudes poses two severe problems: (a) The relative phases of the atomic displacements are lost; and (b) the amplitudes may reflect disorder in the structure and systematic error in the diffraction experiment in addition to motion, but the three contributions cannot be separated on the basis of measurements at a single temperature. Several approximate ways to solve these problems, e.g. rigid-body and segmented-rigid-body analysis, are reviewed together with their limitations. A more recent approach that represents a significant advance with respect to both difficulties is also described: Crystal structures are determined over a range of temperatures; the mean square amplitude quantities are interpreted by taking explicit account of their temperature dependence, i.e. by exploiting the difference in behavior of a microscopic oscillator in the low-temperature, quantum regime and in the high temperature, classical regime. A distinction between low-frequency and high frequency motion, disorder, and systematic error becomes possible with this model; this is illustrated with the help of case studies. PMID- 11031284 TI - Quantitative atom-atom potentials from rotational tunneling: their extraction and their use. AB - Rotational tunneling of small molecular groups has been the subject of considerable theoretical and experimental activity for several decades. Much of this activity has been driven by the promise of exploiting the extreme sensitivity of quantum tunneling to interatomic potentials, but until recently, there was no straightforward means by which quantitative information about these potentials could be extracted. This review explains how a quantitative method, suitable for general application, was developed. It then goes on to show how this has been used to understand tunneling systems for which no previous satisfactory explanation had been found. The application of the methodology, and its results, to other disciplines is discussed. PMID- 11031285 TI - Decoding the dynamical information embedded in highly mixed quantum states. AB - The standard description of the vibrational and rotational motion of polyatomic molecules, as expressed by the distortable rotor/harmonic oscillator approximation, provides an adequate description of the molecular quantum states only in regions of low total state density. When the total state density is large, exceeding 100 states/cm(-1), the vibrational dynamics are "dissipative" and the fundamental process of intramolecular vibrational energy redistribution is operative. The presence of intramolecular vibrational energy redistribution leads to molecular quantum states of a qualitatively different nature. With respect to a normal-mode vibrational basis, these quantum states are "highly mixed" in their vibrational character and represent nuclear motion that is a combination of all the normal-mode motions. This review describes frequency domain spectroscopy techniques designed to probe the vibrational, rotational, and structural composition of these eigenstates. Recent work that investigates spectroscopy between highly mixed states is also reviewed. PMID- 11031286 TI - Large-scale shape changes in proteins and macromolecular complexes. AB - Proteins and RNA undergo intricate motions as they carry out functions in biological systems. These motions frequently entail large-scale conformational changes that induce changes in the surface structure, or shape, of a molecule. This review describes the experimental characterization of large-scale shape changes in proteins and macromolecular complexes and the effects of such changes on macromolecular behavior. We describe several important results that have been obtained by using small-angle scattering, which is emerging as a powerful technique for determining macromolecular shapes and elucidating the quaternary structure of macromolecular assemblies. PMID- 11031287 TI - Reflection absorption infrared spectroscopy and the structure of molecular adsorbates on metal surfaces. AB - Infrared (IR) spectroscopy is widely used to identify molecular adsorbates that form on metals in the course of surface chemical reactions. Because IR spectroscopy is one of the few surface-sensitive probes that provide molecule specific information without perturbing the chemisorbed state, there is great interest in extracting as much structural information from the spectra as possible. The various ways IR spectroscopy is used to determine the structure of molecular adsorbates, from strictly qualitative interpretations based on symmetry selection rules to the use of ab initio electronic structure calculations to predict the IR spectrum of a chemisorbed molecule, are reviewed. PMID- 11031288 TI - The dynamics of noble gas--halogen molecules and clusters. AB - The vibrational relaxation of noble gas-halogen van der Waals clusters has been fertile territory for the development of experimental and theoretical tools for state-to-state dynamics studies. Proceeding through the various combinations of noble gas atoms and halogen molecules, one goes from "simple" direct vibrational predissociation, through sequential relaxation pathways, to the statistical intramolecular vibrational relaxation limit. In some cases the vibrational processes dominate, in others electronically nonadiabatic processes, including chemical reactions, interfere. Thus the noble gas-halogen species provide test cases for most of the dynamical processes available to more "normal" molecules. This review discusses the current state-of-the-art for such studies and points to important problems that remain to be solved. PMID- 11031289 TI - Molecular dynamics simulation of nucleic acids. AB - We review molecular dynamics simulations of nucleic acids, including those completed from 1995 to 2000, with a focus on the applications and results rather than the methods. After the introduction, which discusses recent advances in the simulation of nucleic acids in solution, we describe force fields for nucleic acids and then provide a detailed summary of the published literature. We emphasize simulations of small nucleic acids ( approximately 6 to 24 mer) in explicit solvent with counterions, using reliable force fields and modern simulation protocols that properly represent the long-range electrostatic interactions. We also provide some limited discussion of simulation in the absence of explicit solvent. Absent from this discussion are results from simulations of protein-nucleic acid complexes and modified DNA analogs. Highlights from the molecular dynamics simulation are the spontaneous observation of A B transitions in duplex DNA in response to the environment, specific ion binding and hydration, and reliable representation of protein-nucleic acid interactions. We close by examining major issues and the future promise for these methods. PMID- 11031290 TI - Chemistry and microphysics of polar stratospheric clouds and cirrus clouds. AB - Ice particles found within polar stratospheric clouds (PSCs) and upper tropospheric cirrus clouds can dramatically impact the chemistry and climate of the Earth's atmosphere. The formation of PSCs and the subsequent chemical reactions that occur on their surfaces are key components of the massive ozone hole observed each spring over Antarctica. Cirrus clouds also provide surfaces for heterogeneous reactions and significantly modify the Earth's climate by changing the visible and infrared radiation fluxes. Although the role of ice particles in climate and chemistry is well recognized, the exact mechanisms of cloud formation are still unknown, and thus it is difficult to predict how anthropogenic activities will change cloud abundances in the future. This article focuses on the nucleation, chemistry, and microphysical properties of ice particles composing PSCs and cirrus clouds. A general overview of the current state of research is presented along with some unresolved issues facing scientists in the future. PMID- 11031291 TI - Monte Carlo methods in electronic structures for large systems. AB - Quantum Monte Carlo methods have recently made it possible to calculate the electronic structure of relatively large molecular systems with very high accuracy. These large systems range from positron complexes [NH(2),Ps] with approximately 10 electrons to C(20) isomers with 120 electrons, to silicon crystal structures of 250 atoms and 1000 valence electrons. The techniques for such calculations and a sampling of applications are reviewed. PMID- 11031292 TI - Thermodynamics of the size and shape of nanocrystals: epitaxial Ge on Si(001). AB - The growth and evolution of strained epitaxial Ge on a Si(001) surface provides a rich system for exploring the behavior of strongly interacting nanocrystals. In the temperature regime above 500 degrees C, there are two different (metastable) shapes of defect-free nanocrystals, termed pyramids and domes, that dominate the system depending on the temperature of the substrate during growth and the amount of Ge deposited. In contrast to the usual case considered in nucleation theory, the relaxation of the strain energy at the surface of the nanocrystals makes those surfaces stabilizing, i.e. the surface contribution to the free energy of the Ge nanocrystals is negative. Given that the edges of the nanocrystals are destabilizing (positive free energy), the interaction of the surfaces and edges of the nanocrystals in an ensemble renders an internal free energy for the system that has a local minimum with respect to the size (volume) of the nanocrystal. At finite temperatures, this free energy yields a size distribution with a characteristic centroid, width, and skewness for each nanocrystal shape. The smaller pyramids transform into domes when they grow to the point where they can surmount a kinetic energy barrier between the two structures. However, the Ge nanocrystals also effectively repel one another strongly via the strain fields that are produced in the Si substrate. This repulsive interaction makes the ensemble of Ge nanocrystals a highly nonideal thermodynamic system and, in turn, makes the free energies of the nanocrystals a function of their number density, or equivalently a function of the amount of Ge deposited. The interplay of the stabilizing effect of the nanocrystal surfaces and the destabilizing influence of their repulsive interactions yields a complex behavior for the nanocrystal-size distributions that can nonetheless be modeled using simple thermodynamic expressions. PMID- 11031293 TI - Semiclassical calculation of chemical reaction dynamics via wavepacket correlation functions. AB - Calculation of chemical reaction dynamics is central to theoretical chemistry. The majority of calculations use either classical mechanics, which is computationally inexpensive but misses quantum effects, such as tunneling and interference, or quantum mechanics, which is computationally expensive and often conceptually opaque. An appealing middle ground is the use of semiclassical mechanics. Indeed, since the early 1970s there has been great interest in using semiclassical methods to calculate reaction probabilities. However, despite the elegance of classical S-matrix theory, numerical results on even the simplest reactive systems remained out of reach. Recently, with advances both in correlation function formulations of reactive scattering as well as in semiclassical methods, it has become possible for the first time to calculate reaction probabilities semiclassically. The correlation function methods are contrasted with recent flux-based methods, which, although providing somewhat more compact expressions for the cumulative reactive probability, are less compatible with semiclassical implementation. PMID- 11031294 TI - Self-assembled ceramics produced by complex-fluid templation. AB - This review examines the use of self-assembly in the fabrication of ceramic mesostructures, emphasizing the use of amphiphilic surfactants and block copolymers. The association between this area of research and biomimetics is discussed, linking developments in synthetic self-assembly with biomineralization. The fabrication of hierarchical structures through the use of simultaneous processing is shown to be a necessary condition for applications of this new technology. PMID- 11031295 TI - Theoretical studies of atomic-scale processes relevant to crystal growth. AB - The study of adsorption, diffusion, island formation, and interlayer transport of atoms on a growing surface has been an active field in the past decade, because of both experimental and theoretical advances. Experiments can give detailed images of patterns formed on growing surfaces. An important challenge to the theoretical studies is the identification of dynamical processes controlling the pattern formation and overall surface morphology. This can be achieved by accurate modeling of the atomic interactions, a thorough search for active atomic scale processes, and simulation of the growth on the experimental timescale to allow for detailed comparison with the experimental measurements. An overview of some of the theoretical methodology used in these studies and results obtained for one of the most extensively studied systems, Pt(111), is given here. A remarkable richness of phenomena has emerged from these studies, where apparently small effects can shift the balance between competing molecular processes and thereby change the morphology of a growing surface. The article concludes with a discussion of possible future directions in this research area. PMID- 11031296 TI - New technologies in electron spin resonance. AB - New electron spin resonance (ESR) technologies have been developed, which have led to new and improved applications. (a) The development of two-dimensional Fourier transform (FT) ESR required spectrometers providing intense pi/2 microwave pulses of very short (3-5 ns) duration, wide bandwidths, and very short dead times. It has enabled studies that resolve sophisticated details of molecular dynamics in complex fluids. (b) Methods that produce multiple quantum coherences by pulsed ESR now enable accurate measurements of large distances (>12A). (c) One of the most important advances has been the extension of ESR to high magnetic fields and high frequencies. This has benefited from the utilization of quasi-optical methods, especially above 150 GHz. The greatly improved orientational resolution and the faster "snapshot" of motions that are provided by ESR at high frequencies enhance studies of molecular dynamics. The use of both high and lower frequencies enables one to unravel faster and slower modes from the complex dynamics of fluids and macromolecules. (d) The development of FT-ESR imaging required substantial pulsed field gradients lasting only 50-100 ns. ESR imaging is effective in studying diffusion in fluids. Areas for further development are also described. PMID- 11031297 TI - Multidimensional femtosecond correlation spectroscopies of electronic and vibrational excitations. AB - Femtosecond visible and infrared analogues of multiple-pulse nuclear magnetic resonance techniques provide novel snapshot probes into the structure and electronic and vibrational dynamics of complex molecular assemblies such as photosynthetic antennae, proteins, and hydrogen-bonded liquids. A classical oscillator description of these spectroscopies in terms of interacting quasiparticles (rather than transitions among global eigenstates) is developed and sets the stage for designing new pulse sequences and inverting the multidimensional signals to yield molecular structures. Considerable computational advantages and a clear physical insight into the origin of the response and the relevant coherence sizes are provided by a real-space analysis of the underlying coherence-transfer pathways in Liouville space. PMID- 11031298 TI - Structures and dynamics of molecules on liquid beam surfaces. AB - In this review, we describe experimental studies on structures and dynamics of molecules on clean liquid surfaces in a vacuum. These studies use clean-surface preparations in combination with highly sensitive laser-, photoelectron-, and mass-spectroscopic techniques. In particular, we refer to our recent studies on solvation structures and reactions on various solution surfaces using liquid beam multiphoton ionization-mass spectrometry. These include (a) aggregation of solute molecules on solution surfaces such as formation of solute pairs, clusters, and islands; and (b) chemical reactions induced by photoexcitation and -ionization, such as a nucleophilic reaction of phenyl ketones, polymerization of halo anilines, and reduction of cations by solvated electrons. PMID- 11031299 TI - Effects of high pressure on molecules. AB - Recent high-pressure studies reveal a wealth of new information about the behavior of molecular materials subjected to pressures well into the multimegabar range (several hundred gigapascal), corresponding to compressions in excess of an order of magnitude. Under such conditions, bonding patterns established for molecular systems near ambient conditions change dramatically, causing profound effects on numerous physical and chemical properties and leading to the formation of new classes of materials. Representative systems are examined to illustrate key phenomena, including the evolution of structure and bonding with compression; pressure-induced phase transitions and chemical reactions; pressure-tuning of vibrational dynamics, quantum effects, and excited electronic states; and novel states of electronic and magnetic order. Examples are taken from simple elemental molecules (e.g. homonuclear diatomics), simple heteronuclear species, hydrogen bonded systems (including H2O), simple molecular mixtures, and selected larger, more complex molecules. There are many implications that span the sciences. PMID- 11031300 TI - The cell wall as a barrier to uptake of metal ions in the unicellular green alga Chlamydomonas reinhardtii (Chlorophyceae). AB - The cell walls of plants, including those of algae, have the capacity to bind metal ions in negatively charged sites. The authors had already shown that the wild type (walled) strain of the unicellular green alga Chlamydomonas reinhardtii Dangeard was more tolerant to Cd, Co, Cu, and Ni than a wall-less mutant of the same species. The objective of the present study was to determine if the tolerance to metals was associated with an increased adsorption of the same metals to the cell wall. Adsorbed metal was defined as that fraction that could be removed with a solution containing Na(2)EDTA and CaCl(2). The fraction that remained after the EDTA/CaCl(2) wash was considered to be strongly bound in the cell. When exposed to metals, singly, in solution for 24 h, cells of both strains accumulated the metals. The original hypothesis was supported by the results for Cd, Co, and Ni insofar as significantly higher concentrations of these metals were in the loosely bound fraction of the walled strain in comparison with the wall-less strain. However, there are three reasons why the potentially protective effect of the cell wall did not explain differential tolerance of the two strains. After 24 h of exposure (1) less Cd was accumulated internally by the wall-less strain than by the walled strain, (2) very little of the accumulated Cu was in the loosely bound fraction of the walled strain, and (3) the two strains accumulated comparable and relatively high amounts of internal Cu. Unexpectedly, significant amounts of Cd and Cu were also removable from the surface of the wall less cells. One possible explanation for these apparently externally bound metals in the wall-less strain is that the cells exuded metal-chelating molecules that decreased the ability of metal ions to penetrate the plasma membrane. It was concluded that metal tolerance in this alga must involve a complex of mechanisms involving both internal and external detoxification of metal ions. PMID- 11031301 TI - Quantification of metal bioavailability for lettuce (Lactuca sativa L.) in field soils. AB - Understanding metal bioavailability of plants in soils requires, apart from physiological processes and symbiosis with arbuscular mycorrhizal fungi, the consideration of the chemical availability in the soil solution (the intensity of the toxic exposure) and the soil's capacity to supply the metal (capacity). In this contribution we report on the time-dependent accumulation of As, Cd, Cr, Cu, Ni, Pb, and Zn in lettuce (Lactuca sativa L.). Bioassays with 17 Dutch field soils and two artificially metal-contaminated soils were carried out. Phytotoxicity was observed in soils with pH (pore water) <4.8. Metal uptake is shown to be both metal- and soil-dependent and strongly depends on the amount of water the plant transpired and the available concentration in the water. No net accumulation of As, Pb, Ni, and especially Cr was observed in most soils tested. The latter observation is in agreement with findings of Zayed et al. (Planta, 1998 206:293-299), who reported that translocation of Cr from roots to shoots is extremely limited. Internal Cd levels in the plants varied greatly among soils, whereas plant tissue concentrations of Zn and especially Cu appear to be regulated at more or less fixed levels. The 0.01 M CaCl(2)-extractable metal pool provides the best descriptor for the capacity of the soil to supply Cd and Zn. This enabled the development of models that are suited to predict Zn and Cd uptake by lettuce in both field soils (weathered soils) and soils to which metal salts were added, which is common practice in toxicity testing of chemicals. It is concluded that of all metals included in this study, Cd is the metal of most concern due to bioaccumulation through the soil-plant-animal food chain as Cd is the only metal that might pose human or animal health risks at plant tissue concentrations that are not directly phytotoxic. Finally, application of the models for risk assessment purposes is discussed. PMID- 11031302 TI - Toxicity and bioaccumulation of copper to black bindweed (Fallopia convolvulus) in relation to bioavailability and the age of soil contamination. AB - The use of ecotoxicity test results obtained in the laboratory for prediction of effects of potentially toxic concentrations of chemicals in the field is hampered by several factors differing between the laboratory and the field situations. One important factor is the binding of test chemicals to soil, which is affected by the age of the contamination and soil type. The present study investigated the effect of contamination age by introducing an aging period of 1 to 12 weeks between mixing the test substance, copper sulfate, in with soil and introducing the test plant, Fallopia convolvulus (L.) A. Love. Copper accumulation, emergence of cotyledons, and growth were assessed and related to total (boiling with HNO(3)) and 0.01 M CaCl(2)- and DTPA-extractable soil copper concentrations. Aging of the copper-contaminated soil had only small effects on bioaccumulation of copper, copper toxicity, and extractable soil copper fractions. Soil copper had no effect on emergence of cotyledons. Estimated EC(50) values for shoot and root growth averaged 280 mg Cu/kg. Effects on growth in these laboratory-treated soils were much more severe than in a study performed in soil from an old copper contaminated field site. Neither CaCl(2)- nor DTPA-extractable copper fractions could explain all of the differences in effects between aged spiked soil and field soil. The accumulation pattern for roots and shoots of F. convolvulus indicated that excessive copper was accumulated and adsorbed mainly by the fine roots, whereby the copper concentrations of other plant parts were kept low until the plant was no longer able to maintain this regulation. An internal threshold for effects on growth of about 20 mg Cu/kg shoot dry weight was estimated, coinciding with a soil copper concentration of approximately 200 mg/kg. PMID- 11031303 TI - Assessment of barium toxicity in bush beans. AB - External and internal lowest observed effect concentrations (LOECs) for Ba in bean plants (Phaseolus vulgaris) were established using a nutrient solution culture system where BaSO(4) precipitation in the growth medium was avoided. This was achieved by alternating every 24 h with a nutrient solution containing Ba (0, 50, 500, or 5000 microM) and all essential elements except S, with another containing S and all other nutrients but no Ba. The external LOEC for acute toxicity symptoms in the form of leaf withering and leaf growth inhibition was 481 microM of free Ba(2+). This was also the LOEC for the complete inhibition of elongation of secondary roots, while for the elongation of the primary root the external LOEC was 4,821 microM. Barium interfered with both the sulfate transport from roots to shoots and the import of Ca into leaves. However, K was the most Ba sensitive nutrient. External LOEC for reduced leaf K concentrations was 48 microM free Ba(2+); the corresponding internal LOECs for primary and trifoliolate leaves were 700 and 460 mg kg(-1) DW, respectively. PMID- 11031304 TI - Toxicity and bioavailability of copper herbicides (Clearigate, Cutrine-Plus, and copper sulfate) to freshwater animals. AB - In designing aquatic herbicides containing copper, an important goal is to maximize efficacy for target species while minimizing risks for nontarget species. To have a margin of safety for nontarget species, the concentration, duration of exposure (i.e., uptake), and form (i.e., species) of copper used for herbicidal properties should not elicit adverse effects on populations of nontarget species. To determine the potential for risk or adverse effects (conversely the margin of safety), data regarding the comparative toxicity of copper-containing herbicides are crucial. A series of comparative toxicity experiments was conducted, including baseline estimates of toxicity (LC50s, LOECs), sensitive species relationships (thresholds and exposure-response slopes), and bioavailability of toxic concentrations and forms of copper 7 days after initial herbicide application. Aqueous 48-h toxicity experiments were performed to contrast responses of Daphnia magna Strauss, Hyalella azteca Saussure, Chironomus tentans Fabricius, and Pimephales promelas Rafinesque to copper herbicides: Clearigate(R), Cutrine(R)-Plus, and copper sulfate. D. magna was the most sensitive aquatic animal tested for all three herbicides; 48-h LC50s for organisms exposed to Clearigate, Cutrine-Plus, and copper sulfate were 29.4, 11.3, and 18. 9 microg Cu/L, respectively. In terms of potency (calculated from the linearized portion of the exposure-response curves, which included 50% mortality), D. magna was the most sensitive animal tested. Organisms exposed to Clearigate, Cutrine-Plus, and copper sulfate had exposure-response slopes of 2.55, 8.61, and 5.07% mortality/microg Cu/L, respectively. Bioavailability of Clearigate and Cutrine-Plus was determined by comparing survival data (LC50s) of test organisms exposed to herbicide concentrations during the first and last 48-h of a 7-day exposure period. Even in these relatively simplified water-only exposures, a transformation of copper to less bioavailable species over time was observed with a 100-200% decrease in toxicity (i.e., an increase in 48-h LC50s) for all four test animals. This series of laboratory experiments provides a worst case scenario for determining the risk associated with the manufacturer's recommended application rates of Clearigate (100-1,000 microg Cu/L), Cutrine-Plus (200-1,000 microg Cu/L), and copper sulfate (100-500 microg Cu/L) in natural waters for four nontarget freshwater animals. PMID- 11031305 TI - An evaluation of the toxicity of contaminated sediments from Waukegan Harbor, Illinois, following remediation. AB - Waukegan Harbor in Illinois was designated as a Great Lakes Area of Concern due to high concentrations of sediment-associated polychlorinated biphenyls (PCBs). The objective of this study was to evaluate the toxicity of 20 sediment samples collected after remediation (primarily dredging) of Waukegan Harbor for PCBs. A 42-day whole sediment toxicity test with the amphipod Hyalella azteca (28-day sediment exposure followed by a 14-day reproductive phase) and sediment toxicity tests with Microtox(R) were conducted to evaluate sediments from Waukegan Harbor. Endpoints measured were survival, growth, and reproduction (amphipods) and luminescent light emission (bacteria). Survival of amphipods was significantly reduced in 6 of the 20 sediment samples relative to the control. Growth of amphipods (either length or weight) was significantly reduced relative to the control in all samples. However, reproduction of amphipods identified only two samples as toxic relative to the control. The Microtox basic test, conducted with organic extracts of sediments identified only one site as toxic. In contrast, the Microtox solid-phase test identified about 50% of the samples as toxic. A significant negative correlation was observed between reproduction and the concentration of three polynuclear aromatic hydrocarbons (PAHs) normalized to total organic carbon. Sediment chemistry and toxicity data were evaluated using sediment quality guidelines (consensus-based probable effect concentrations, PECs). Results of these analyses indicate that sediment samples from Waukegan Harbor were toxic to H. azteca contaminated at similar contaminant concentrations as sediment samples that were toxic to H. azteca from other areas of the United States. The relationship between PECs and the observed toxicity was not as strong for the Microtox test. The results of this study indicate that the first phase of sediment remediation in Waukegan Harbor successfully lowered concentrations of PCBs at the site. Though the sediments were generally not lethal, there were still sublethal effects of contaminants in sediments at this site observed on amphipods in long-term exposures (associated with elevated concentrations of metals, PCBs, and PAHs). PMID- 11031306 TI - Sediment toxicity assessment: comparison of standard and new testing designs. AB - Standard methods of sediment toxicity testing are fairly well accepted; however, as with all else, evolution of these methods is inevitable. We compared a standard ASTM 10-day amphipod toxicity testing method with smaller, 48- and 96-h test methods using very toxic and reference sediments. In addition we compared parallel exposures of single species, either the amphipod Ampelisca abdita or the mysid Americamysis bahia, to multiple species, mysid, and amphipod, cohabiting the same types of chambers. These comparisons were performed for both water-only and sediment-water tests. Results of the comparison of the standard ASTM 10-day amphipod test with the smaller, 48- and 96-h test chambers indicate that survival was high in both test designs using the reference sediment. With toxic sediments, complete mortality occurred in less than 48 h using the smaller experimental chambers and only after 96 h in the larger experimental chambers. We concluded that although time to death is shorter in the smaller, shorter exposure chambers, there was no overall change in mortality for the organisms, and that the smaller chambers were predictive of the results obtained with larger chambers and longer exposures. For multiple species testing in whole sediment exposures there was no change in toxicity to either the amphipod or the mysid when they cohabited the same chamber. In contrast, for water-only exposures, A. bahia demonstrated less sensitivity when cohabiting the same chamber as A. abdita. Therefore, during whole sediment testing we can add A. bahia and A. abdita to the same test chamber without changing the toxicity to either species; however, in our 10-ml water-only exposures, the species should be tested separately. PMID- 11031307 TI - A comparison of selected diversity, similarity, and biotic indices for detecting changes in benthic-invertebrate community structure and stream quality. AB - Implementation of advanced wastewater treatment at the two municipal wastewater treatment plants for Indianapolis, Indiana, resulted in substantial improvement in the quality of the receiving stream and significant changes in the benthic invertebrate community. Diversity, similarity, and biotic indices were compared to determine which indices best reflected changes in the composition of the biota in the river. None of the indices perfectly reflected the changes in river quality or community structure. Similarity indices, especially percentage similarity, exhibit the most promise of the three classes of indices. Diversity indices were least useful, wrongly indicating that water quality deteriorated after the upgrade of the wastewater-treatment plants. The most descriptive tool in analyzing the data was the percentage of Ephemeroptera, Plecoptera, and Trichoptera (EPT) taxa present. Using a mixture of indices and other analytical tools, such as EPT, in the analysis of biological data will ensure the most effective investigations of water quality. PMID- 11031308 TI - Hemocytic lysosome response in the snail Helix aspersa after exposure to the fungicide copper oxychloride. AB - This study investigated the use of lysosomal responses of hemocytes of the common garden snail, Helix aspersa, as biomarker of stress due to exposure to the fungicide copper oxychloride. The neutral red retention (NRR) time assay was employed for this purpose. Two groups of snails were exposed to 80 microg g(-1) and 240 microg g(-1) copper oxychloride in their food, respectively, for a period of 6 weeks. They were compared with a control group to which no copper oxychloride was added. The two groups exposed to the fungicide exhibited significantly higher (p < 0.001) whole body copper concentrations (200.85 +/- 53.5 and 272.24 +/- 67.15 microg g(-1) dry mass, respectively), and significantly shorter (p < 0.001) NRR times (10.22 +/- 3.53 and 2.67 +/- 2.83 min, respectively), after 6 weeks, compared to the control group (67.85 +/- 31.08 microg g(-1) dry mass and 24.44 +/- 8.35 min). In both exposure groups NRR times became progressively shorter as body copper concentrations increased over time. Thus, both exposure concentration and exposure time of copper oxychloride were shown to be important factors influencing lysosomal responses (and therefore NRR times) of H. aspersa hemocytes. It was concluded that these responses in this species, as measured by the NRR time assay, could be considered a useful cellular biomarker of stress resulting from exposure to copper oxychloride. PMID- 11031309 TI - Effect of single and binary combinations of plant-derived molluscicides on reproduction and survival of the snail Achatina fulica. AB - The effects of sublethal treatments (20% and 60% of LC(50)/24 h) with plant derived molluscicides on the reproduction of the giant African snail Achatina fulica were studied. Azadirachta indica oil, Cedrus deodara oil, Allium sativum bulb powder, and Nerium indicum bark powder singly and binary combinations on reproduction and survival of A. fulica were investigated. Repeated treatment occurred on day 0, day 15, and day 30. These plant-derived molluscicides significantly reduced fecundity, egg viability, and survival of A. fulica within 15 days. Discontinuation of the treatments after day 30 did not lead to a recovery trend in the next 30 days. Day 0 sublethal treatment of all the molluscicides caused a maximum reduction in protein, amino acid, DNA, RNA, and phospholipid levels and simultaneous increase in lipid peroxidation in the ovotestis of treated A. fulica. It is believed that sublethal exposure of these molluscicides on snail reproduction is a complex process, involving more than one factor in reducing the reproductive capacity of A. fulica. PMID- 11031310 TI - Combined effects of UV-B, nitrate, and low pH reduce the survival and activity level of larval cascades frogs (Rana cascadae). AB - We investigated interactions between low pH, high nitrate level, and ultraviolet B (UV-B) light on the survival and activity level of larval Cascades frogs (Rana cascadae). We used a fully factorial experimental design, with pH levels of 5 and 7; initial "pulse" nitrate exposure levels of 0, 5, and 20 mg/L; and UV-B present or absent. After a 3-week laboratory exposure, we measured survival and activity level of the larvae. The experiment was repeated two times, in two separate years. Similar effects on survival and activity level were observed in both experiments. R. cascadae survival was not significantly reduced in treatments with individual factors alone (i.e., UV-B control, pH 5 control, or high nitrate level without pH or UV-B). However, in experiments from both years, survival and activity levels of larval R. cascadae were significantly reduced in the treatment with low pH, high nitrate, and UV-B together. In both years, analysis of variance (ANOVA) indicated that pH and nitrate had the greatest effect on survival and that UV-B and nitrate had the greatest effect on activity level. Additional effects were observed in the 1998 experiment on both survival and activity level. In 1998, UV radiation and the interaction term between pH and nitrate (pH x nitrate) had a significant effect on survival. Also in the 1998 experiment, activity level was significantly reduced in treatments at neutral pH with UV, at initial nitrate doses of 5 and 20 mg/L and at neutral pH without UV at an initial nitrate dose of 20 mg/L. We suggest that the adverse effects were due to the multiple stressors acting together. PMID- 11031311 TI - Chlorinated hydrocarbon concentrations in plasma of the Lake Erie water snake (Nerodia sipedon insularum) and northern water snake (Nerodia sipedon sipedon) from the Great Lakes basin in 1998. AB - From the Great Lakes basin, concentrations of 59 congener-specific polychlorinated biphenyls (PCBs) and 14 organochlorine pesticides were measured in blood plasma of northern water snake (Nerodia sipedon sipedon) and Lake Erie water snake (Nerodia sipedon insularum), which is endangered in Canada. In 1998, four male adult Lake Erie water snakes were sampled from Pelee Island, western Lake Erie; four male northern water snakes were sampled at Little Lake, about 20 km north of Parry Sound in central Ontario; and four adult gravid female northern water snakes were sampled from Garden Island, eastern Lake Ontario. The blood plasma was pooled by site for a total of three samples analyzed. The Pelee Island sample from male Lake Erie water snakes contained less than half the lipid concentration (0.349%) than samples from the other sites, but it was the most contaminated with PCBs, even on a wet weight basis. Summed concentration of individual PCBs in the Pelee Island sample was 167 ng/g (wet weight), which was 14-fold higher than the next most contaminated sample, which was from Little Lake. The plasma sample from Little Lake contained 12 ng/g (WW) and was four times more contaminated with PCBs than the sample from female snakes from Garden Island, Lake Ontario. Organochlorine pesticide concentrations in plasma were relatively similar among sites. None of the pesticides was found above trace concentrations (0.1-0.9 ng/g) except pp'-DDE, which occurred at 2-5 ng/g among sites. PCB congener patterns in the Lake Erie water snakes were compared to PCB patterns in plasma of common snapping turtle (Chelydra serpentina serpentina) from Lake Ontario, herring gull eggs (Larus argentatus) from western Lake Erie, and mudpuppy eggs (Necturus maculosus) from the Detroit River. The PCB patterns in water snake and herring gull sample were most similar, followed by the pattern in snapping turtle plasma. The presence of more lower-chlorinated chlorobiphenyls in the mudpuppy eggs relative to the other species made this sample distinct from the water snake, gull, and turtle. PMID- 11031312 TI - Reclamation of wastewater for polyculture of freshwater fish: bioassays using Chlorella and Gambusia. AB - The feasibility of using the secondary effluents from two sewage treatment plants in Hong Kong (Yuen Long and Shek Wu Hui) for fish culture was assessed. Total ammonia contents in the two sewage effluents surpassed the level of 2 mg L(-1) recommended for wastewater fish culture. The two sewage effluents both needed essential elements for supporting algal Chlorella vulgaris growth, whereas only sewage effluent from Yuen Long had contaminants at toxic levels. Total ammonia in water explained more than 80% variations of toxicity of water samples to Gambusia patruelis, mosquito fish, according to regression analysis. Removal of ammonia from the sewage effluent is necessary before being used for fish culture. In addition, the rather high levels of heavy metals (Cu, Zn, Cd) in the effluent should be closely monitored. PMID- 11031313 TI - Metal analysis of scales taken from Arctic grayling. AB - This study examined concentrations of metals in fish scales taken from Arctic grayling using laser ablation-inductively coupled plasma mass spectrometry (LA ICPMS). The purpose was to assess whether scale metal concentrations reflected whole muscle metal concentrations and whether the spatial distribution of metals within an individual scale varied among the growth annuli of the scales. Ten elements (Mg, Ca, Ni, Zn, As, Se, Cd, Sb, Hg, and Pb) were measured in 10 to 16 ablation sites (5 microm radius) on each scale sample from Arctic grayling (Thymallus arcticus) (n = 10 fish). Ca, Mg, and Zn were at physiological levels in all scale samples. Se, Hg, and As were also detected in all scale samples. Only Cd was below detection limits of the LA-ICPMS for all samples, but some of the samples were below detection limits for Sb, Pb, and Ni. The mean scale concentrations for Se, Hg, and Pb were not significantly different from the muscle concentrations and individual fish values were within fourfold of each other. Cd was not detected in either muscle or scale tissue, whereas Sb was detected at low levels in some scale samples but not in any of the muscle samples. Similarly, As was detected in all scale samples but not in muscle, and Ni was detected almost all scale samples but only in one of the muscle samples. Therefore, there were good qualitative and quantitative agreements between the metal concentrations in scale and muscle tissues, with LA-ICPMS analysis of scales appearing to be a more sensitive method of detecting the body burden of Ni and As when compared with muscle tissue. Correlation analyses, performed for Pb, Hg, and Se concentrations, revealed that the scale concentrations for these three metals generally exceeded those of the muscle at low muscle concentrations. The LA-ICPMS analysis of scales had the capability to resolve significant spatial differences in metal concentrations within a fish scale. We conclude that metal analysis of fish scales using LA-ICPMS shows considerable promise as a nonlethal analytical tool to assess metal body burden in fish that could possibly generate a historic record of metal exposure. However, comprehensive validation experiments are still needed. PMID- 11031314 TI - Development of an index of biotic integrity for the Little Arkansas River Basin, Kansas. AB - An index of biotic integrity (IBI) was developed for the Little Arkansas River Basin (LARB) in south-central Kansas by establishing a reference condition for the watershed. Twelve metrics, in six categories, were chosen for use in the IBI. Fish assemblages from 30 sites were selected to represent the highest quality sites (reference sites) remaining in the LARB. In addition, 20 historical sites were used to show changes in the watershed over the last century. The modified IBI was then tested at 10 sites within the basin to assess the affects of urban and agricultural disturbances on fish community structure in the Wichita area. IBI scores were statistically lower for the urban versus the agricultural sites. Overall, IBI scores rated from poor to fair, supporting the contention that the fish communities within the LARB are impaired. PMID- 11031315 TI - Assessment of water quality in Wichita, Kansas, using an index of biotic integrity and analysis of bed sediment and fish tissue for organochlorine insecticides. AB - This study investigated water quality of the Arkansas River and its tributaries in the Wichita, Kansas, area. Water quality assessment included an evaluation of urban versus agricultural areas using a twofold approach: (1) assessment of fish community health using an index of biotic integrity (IBI); and (2) analysis of organochlorine (OC) insecticides in fish tissue (Cyprinus carpio) and sediment samples via gas chromatography. Results of the IBI analysis indicated that the overall health of the fish community was poor. No significant difference existed among IBI scores at urban sites compared to agricultural sites; however, a pattern was noted with higher IBI scores at the agricultural sites. Total IBI scores and individual metrics were not statistically correlated to either normalized or non-normalized organochlorine concentrations in fish or sediment. Lindane, chlordane, 4,4'-DDT, and 4,4'-DDE were the most commonly detected OCs in sediment and fish tissue and often exceeded federal guidelines. Total OC concentrations and their detection frequency were significantly greater in urban than agricultural areas. In addition, a significant correlation was found among total OC concentrations in fish and sediment. This assessment concluded that overall water quality in the Wichita area was poor and that the urban area has significantly impacted water quality. PMID- 11031316 TI - Can excrement and feathers of nestling songbirds be used as biomonitors for heavy metal pollution? AB - Although birds have been frequently used as indicators of heavy metal pollution, few studies have examined pollutant levels in nestling passerines. In this paper we determined the levels of two essential (zinc and copper) and three nonessential heavy metals (lead, cadmium, and arsenic) in the excrement and feathers of great (Parus major) and blue tit (Parus caeruleus) nestlings at a polluted site (near a metallurgic factory) and a reference site (4 km farther east). The excrement of both great and blue tit nestlings contained significantly higher concentrations of arsenic, cadmium, copper, and lead at the polluted site. Zinc concentrations did not differ significantly between sites for both species. The feathers of great and blue tit nestlings accumulated significantly higher concentrations of lead at the polluted site than at the reference site. Zinc levels in the feathers of great tit nestlings were significantly higher at the reference site than at the polluted site. For all other elements considered, concentrations did not differ significantly between the two sites. There were no interspecific differences in metal levels between great and blue tits in both excrement and feathers. There was a significant positive correlation between the lead concentration in the excrement and feathers for both great and blue tits. We therefore conclude that excrement of great and blue tit nestlings can be used as a biomonitor for heavy metals (lead, cadmium, arsenic, and copper), whereas feathers appear only to be suitable as a biomonitor for lead pollution. PMID- 11031317 TI - How can we use the red fox as a bioindicator of organochlorines? AB - The exposure of red foxes to hexachlorobenzene (HCB), dichlorodiphenyltrichloroethan (DDT), DDT derivatives, and polychlorinated biphenyls (PCBs) in central Italy was determined by analyzing muscle and adipose tissue samples from 57 red foxes collected in 1992-1993 in three areas of Tuscany, each characterized by different levels of anthropization. Chemical analysis showed that HCB occurred in low concentrations, ranging from an average of 0.08 microg/g lipid basis in fat of foxes from Pisa, the least agricultural area, to 1.06 microg/g in muscle of those from Grosseto, the least industrialized area. DDTs were calculated as the sum of the op' and pp'DDT isomer concentrations and the DDT derivatives DDE and DDD. The lowest values of DDTs were found in foxes from Pisa, with mean values in muscle of 0.55 microg/g and 0.14 microg/g in fat. In the outskirts of Siena, the corresponding values were 1.16 microg/g and 0.49 microg/g. Higher values were found in the Grosseto area: 4.17 microg/g in muscle and 0.67 microg/g in fat. The most abundant DDT was pp'DDE in all areas, constituting more than half of total DDTs. Mean and standard deviations of PCB concentrations (calculated as the sum of the 31 identified and quantified congeners), were 20.2 microg/g in muscle and 7.2 microg/g in fat of red foxes from the Siena area. Values in the Pisa area were lower (7. 6 microg/g in muscle and 1.8 microg/g in fat), whereas the highest levels were found in the Grosseto area (38 microg/g in muscle and 7. 5 microg/g in fat). PCB fingerprints showed that hexa- and heptachlorobiphenyls were the most common congeners, although amounts differed between the three localities and the two tissues considered. As far as PCB composition is concerned, detailed observation revealed that although the three areas highly differed between each other, the most abundant congeners were the 22'344'5' (PCB 138) and 22'44'55' (PCB 153) hexachlorbiphenyls and 22'33'44'5 (PCB 170) and 22'344'55' (PCB 180) heptachlorobiphenyls; these accounted for over 60% of total PCB residues in muscle and fat. Concentrations of the remaining PCB congeners were all lower than 5% of total PCBs, with the exception of octachlorobiphenyl 22'33'44'55' (PCB 194) in fat tissue. Foxes may be suitable biological indicators to detect organochlorine contamination in the environment because of their catholic food habits. This information may be used to select specific bioindicators for assessing the relative level of pollutants in that area. PMID- 11031319 TI - Reflections. PMID- 11031318 TI - Biochemical and ultrastructural alterations caused by newly synthesized 1,2,4 triazole[1,5a]pyrimidine derivatives against Phytomonas staheli (Trypanosomatidae). AB - Six compounds, all newly synthesized triazole-pyrimidine derivatives that proved inhibitory of in in vitro growth of epimastigotes in Trypanosoma cruzi and of promastigotes of Leishmania donovani and Phytomonas staheli, were studied to investigate their toxic effects. As a biological model, the plant trypanosome P. staheli, which causes sudden wilt in the oil palm and Hartrot in the coconut palm, was used. The six compounds markedly inhibited macromolecule synthesis (nucleic acids and proteins) by the parasite. The cells treated with these compounds present severe damage in their ultrastructure-intense 'vacuolization, and appearance of lysosomes as well as other residual bodies. The mitochondrial section appeared larger in size. with a swollen matrix. In addition, these compounds changed the excretion of end metabolites, primarily affecting ethanol and acetate excretion, possibly by directly influencing certain enzymes (alcohol dehydrogenase and acetate synthetase) or their synthesis. 2000 Elsevier Science Ltd. PMID- 11031320 TI - Simulation of automatic measurement of cell distribution inhomogeneity in clonogenic assays. AB - The use of hematopoietic progenitors in toxicology often requires scoring colonies, micro- and macroclusters, and evaluation of their distribution while taking into account their shape. Unfortunately, this evaluation is long and tedious, and classification errors may occur. Image analysis can improve the method if there is an automatic way of performing the classification analysis. It has been previously demonstrated that cell conglomerates can be automatically detected. The present paper describes three image analysis algorithms and reports on their capabilities to analyze cell heterogeneity in culture. The comparative tests are made on computer-generated images representing different cell distributions. Combining two of these three algorithms gives fairly good results on complex images created by adding various simple distributions. PMID- 11031321 TI - Il-1-related cytokine responses of nonimmune skin cells subjected to CEES exposure with and without potential vesicant antagonists. AB - Sulfur mustard provokes an acute inflammatory response in skin. To determine if keratinocytes regulate this response and whether three potential vesicant antagonists can counteract adverse changes, specimens of EpiDerm (MatTek Corp., Ashland, MA), a human skin model of differentiating keratinocytes, were exposed 2 h to humidified air with or without 2-chloroethyl ethyl sulfide (CEES, 1.72-1.73 mg/L/min) with or without 10 mM niacinamide, a poly (ADP-ribose) polymerase (PARP) inhibitor, 25 microM CGS9343B (calmodulin antagonist), or 8.4 mM leupeptin (cysteine protease inhibitor). After a 22-h incubation, levels of interleukin-1 alpha (IL-1alpha), its receptor antagonist (IL-1Ra), soluble type II receptor (sIL-1RII) and prostaglandin-E(2) (PGE(2)) were determined. Methylthiazole tetrazolium (MTT) viability tests and histological observations were also conducted. PGE(2) levels were abundant but unaffected by CEES regardless of antagonist presence. Total amounts (media plus lysate) of IL-1alpha, IL-1Ra, and sIL-1RII were reduced with CEES irrespective of antagonist. CEES promoted the release of IL-1Ra. Exposure of EpiDerm to CEES in the presence of the vesicant antagonists did not improve viability or counteract histological damage. We conclude CEES depresses total IL-1alpha and related cytokines, does not affect PGE(2) release, and adverse changes associated with CEES-exposed EpiDerm are not ameliorated by these particular antagonists. Dramatically increased (5- to 10 fold) release of IL-1Ra may provide a useful marker for cytotoxicity. The high level of IL-1Ra and increased release with injury suggest a primary function in down-regulating IL-1 inflammatory responses in skin. PMID- 11031323 TI - Effects of particulate and soluble cadmium species on biochemical and functional parameters in cultured murine macrophages. AB - Cultured murine macrophages (RAW 264.7) were used to evaluate the temporal relationships between cytotoxicity, phagocytosis, tumor necrosis factor-alpha (TNF-alpha), and nitric oxide (NO) production, and alterations in expression of stress proteins after exposure to cadmium oxide (CdO) or cadmium chloride (CdCl(2)), particulate and soluble forms of cadmium, respectively. Macrophages were exposed in vitro to CdO (25 or 50 microg) or CdCl(2) (30 or 40 microM) for 2 to 72 h. Cytotoxicity was not evident until 18 h when exposed to 30 microM CdCl(2) or 25 microg CdO, but occurred as early as 12 h after exposure to 40 microM CdCl(2) or 50 microg CdO. Relative to untreated controls, phagocytic activity decreased progressively from 2 to 24 h after exposure to both forms of cadmium. TNF-alpha levels increased to 2- to 3-fold after 4 h and remained elevated until 24 h after exposure to 25 and 50 microg CdO and 30 microM CdCl(2), but decreased by 18-24 h at 40 microM CdCl(2). CdCl(2) or CdO alone did not induce NO; however, both cadmium species reduced lipopolysaccharide (LPS) stimulated NO production in a dose-dependent manner. Enhanced de novo synthesis of 70- and 90-kD heat shock, or stress, proteins was observed 2 to 8 h after exposure to both CdCl(2) and CdO; however, synthesis of these proteins returned to control levels by 24 h. Stress protein synthesis was enhanced by CdCl(2) or CdO prior to cytotoxicity, but coincided with a decrease in phagocytic capacity and an increase in TNF-a levels. The data suggest that cultured macrophages respond similarly in vitro to a particulate form and a soluble form of cadmium in a cell type that plays a pivotal role in inflammatory and immune responses. PMID- 11031324 TI - In vitro toxicity of mercury, cadmium, and arsenic to platelet aggregation: influence of adenylate cyclase and phosphodiesterase activity. AB - In vitro effect of mercury (Hg2+), cadmium (Cd2+), and arsenic (As3+) on adenylate cyclase (AC) and phosphodiesterase (PDE) activity in relation to platelet aggregation (PA) was studied in rats. Cd(2+) significantly elevated cAMP (p < 0.005) in a dose-dependent (5, 10 and 20 pmoles) manner while Hg(2+) and As(3+) significantly reduced the cAMP level (p < 0.01 and p < 0.005, respectively). Our studies further reveal that Hg21 and As(3+) inhibit AC and stimulate PDE activity with a concomitant increase in the rate of PA. On the other hand, Cd(2+) stimulates AC and inhibits PDE activity with a decrease in the rate of PA. The present investigation suggests that cellular cAMP is a regulatory molecule in the event of PA and the disruption of its homeostasis is directly correlated to xenobiotic effects on PA. It is concluded that other than divalent heavy metal cations, As(3+) appears to be one of the most toxic xenobiotics to platelet function. PMID- 11031322 TI - Contact sensitizers specifically increase MHC class II expression on murine immature dendritic cells. AB - Contact sensitivity is a T-cell-mediated immune disease that can occur when low molecular-weight chemicals penetrate the skin. In vivo topical application of chemical sensitizers results in morphological modification of Langerhans cells (LC). Moreover, within 18 h, LC increase their major histocompatibility complex (MHC) class II antigens expression and migrate to lymph nodes where they present the sensitizer to T lymphocytes. We wanted to determine if such an effect could also be observed in vitro. However, because of the high genetic diversity encountered in humans, assays were performed with dendritic cells (DC) obtained from a Balb/c mouse strain. The capacity of a strong sensitizer, DNBS (2,4 dinitrobenzene sulfonic acid), to modulate the phenotype of bone marrow-derived DC in vitro, was investigated. A specific and marked increase of MHC class II molecules expression was observed within 18 h. To eliminate the use of animals in sensitization studies, the XS52 DC line was tested at an immature stage. A 30-min contact with the strong sensitizers DNBS and oxazolone, or the moderate mercaptobenzothiazole, resulted in upregulation of MHC class II molecules expression, analyzed after 18-h incubation. This effect was not observed with irritants (dimethyl sulfoxide and sodium lauryl sulfate) nor with a neutral molecule (sodium chloride). These data suggested the possibility of developing an in vitro model for the identification of the sensitizing potential of chemicals, using a constant and non animal-consuming material. PMID- 11031325 TI - Effect of zinc chloride on GSH synthesis rates in various lung cell lines. AB - Zinc toxicity has been linked to decreased reduced glutathione (GSH) and increased oxidized glutathione (GSSG) contents, which might be caused by a GSSG reductase inhibition by zinc. In this study we investigated zinc effects on GSH synthesis rates in various lung cell lines by thin-layer chromatography after (35)S-cysteine incorporation. Two alveolar epithelial cell lines (A549 and L2) and two human fibroblast-like lung cell lines (11Lu and 16Lu) were used in this study. Equipotent protein synthesis inhibition for the different cell lines was reached after 2 h (L2, 11Lu), 3 h (16Lu), and 4 h (A549) zinc exposure (15-200 microM) to cells. Here GSH depletion and GSSG increase in A549 cells were markedly lower than in the other cell lines tested. Incorporation of cysteine (Cys) into GSH was not different in the cell lines tested, while 11Lu cells only demonstrated a decrease of newly synthesized GSH after 1 h of (35)S-Cys exposure when cells were exposed to zinc. Only 11Lu cells showed a markedly decreased Cys availability as compared with the other cell lines. In all cell lines the availability of Cys was not affected by exposure to zinc. No compensating increase in GSH synthesis rates was found after zinc-mediated cellular GSH depletion. PMID- 11031326 TI - CAATSCAN: a review of alternatives. News and information. PMID- 11031328 TI - Alternative medicine for allergy and asthma. AB - Orthodox medical approaches to asthma and allergic respiratory diseases are provided in guidelines developed by professional societies and national or state organizations that represent organized medicine. Alternative therapies may include such orthodox medical therapies as obsolescent formerly used agents, unusual but accepted agents, and agents that are in favor for orthodox therapy in other countries. However, the current growth of complementary and alternative medicine is based on the use of nonorthodox remedies that are becoming increasingly popular with patients and that should be familiar to physicians. Asthma and allergies are frequently treated with such remedies by patients, either as part of self-therapy or on the advice of a complementary and alternative medicine practitioner. The most popular alternative medical treatments are herbs (Western and Asiatic), acupuncture, various types of body manipulation, psychologic therapies, homeopathy, and unusual allergy therapies. There is little evidence in favor of most of these unorthodox treatments, although they are very often reported on favorably by patients. The published evidence that might support some alternative medical practices is reviewed so as to help physicians select alternatives that could appropriately be integrated into orthodox practice. PMID- 11031329 TI - Frozen objects: small airways, big breaths, and asthma. AB - Airway hyperresponsiveness is one of the cardinal features of asthma but remains largely unexplained. The new concept of perturbed myosin binding within airway smooth muscle sheds light on the question of why airway narrowing is limited in the healthy lung and not in the asthmatic lung and points to unanticipated mechanisms through which lung development and allergic status may be major modulators of airway hyperresponsiveness. PMID- 11031330 TI - Monoclonal antibody assays for allergens: pick your antibodies with care! PMID- 11031331 TI - Alternative and complementary medicine: a source of improved therapies for asthma? A challenge for redefining the specialty? PMID- 11031332 TI - Added relief in the treatment of acute recurrent sinusitis with adjunctive mometasone furoate nasal spray. The Nasonex Sinusitis Group. AB - BACKGROUND: Intranasal glucocorticoids are effective in the treatment of allergic rhinitis. Their effectiveness as an anti-inflammatory adjunct in the treatment of acute recurrent sinusitis has not been adequately established in a controlled clinical study. OBJECTIVE: The purpose of this study was to test the hypothesis that intranasal corticosteroid treatment produces additional relief in the treatment of acute sinusitis with oral antibiotics. METHODS: Patients who were 12 years old and older with a history of recurrent sinusitis were treated while experiencing a new episode of acute sinusitis, which was diagnosed by symptoms and confirmed by computed tomography scan of the paranasal sinuses. Patients were treated for 21 days with amoxicillin clavulanate potassium and randomized to receive concurrent mometasone furoate nasal spray (MFNS; Nasonex [400 microg, twice daily]; n = 200 patients) or placebo spray (twice daily; n = 207 patients). Symptom scores for headache, facial pain, congestion, purulent rhinorrhea, postnasal drip, and cough were recorded at baseline and throughout treatment. RESULTS: Baseline symptom scores showed a moderate level of symptom severity comparable in both groups. Patient-recorded twice daily symptom scores showed that adjunctive treatment with MFNS caused a significantly greater decrease in total symptom score (primary efficacy variable) and in individual scores of inflammatory symptoms associated with the obstruction process (headache, congestion, and facial pain) compared with placebo. Symptoms associated with the secretory processes were improved to a lesser degree. Therapy-related local adverse events were not significantly different between groups. CONCLUSION: The addition of intranasal corticosteroid, MFNS 400 microg twice daily, to antibiotics significantly reduces symptoms of acute sinusitis compared with antibiotic treatment alone. PMID- 11031333 TI - A comparison of exhaled nitric oxide and induced sputum as markers of airway inflammation. AB - BACKGROUND: Exhaled nitric oxide (ENO) has been proposed as a noninvasive marker of airway inflammation in asthma. OBJECTIVE: We investigated the relationships among ENO, eosinophilic airway inflammation as measured by induced sputum, and physiologic parameters of disease severity (spirometry and methacholine PC(20)). We also examined the effect of corticosteroid treatment and atopy on ENO levels and eosinophil counts in induced sputum. METHODS: Measurements were taken on one day in 22 healthy nonatopic subjects, 28 healthy atopic subjects, 38 asthmatic subjects not taking inhaled steroids, 35 asthmatic subjects taking inhaled steroids, and 8 subjects with eosinophilic bronchitis without asthma. RESULTS: ENO levels showed significant but weak correlations with eosinophil differential counts in the steroid-naive asthmatic and healthy atopic groups (r (s) < 0.05). ENO levels were significantly lower in the asthmatic subjects taking steroids compared with the asthmatic subjects not taking steroids, despite there being no difference in the sputum cell counts, and a tendency to increased airflow limitation. ENO levels and sputum eosinophil counts were equally good at differentiating from steroid-naive asthmatic subjects. ENO levels were consistently raised in subjects with eosinophilic bronchitis without asthma. Atopy had no effect on ENO levels in the healthy subjects. CONCLUSION: We conclude that ENO is likely to have limited utility as a surrogate clinical measurement for either the presence or severity of eosinophilic airway inflammation, except in steroid-naive subjects. PMID- 11031334 TI - Exhaled nitric oxide levels correlate with measures of disease control in asthma. AB - BACKGROUND: Asthma guidelines emphasize maintaining disease control. However, objective measures of asthma disease control are lacking. OBJECTIVE: We sought to examine the relationship between exhaled nitric oxide (NO) levels and measures of asthma disease control versus asthma disease severity. METHODS: We performed a cross-sectional study of 100 patients (age range, 7-80 years) with asthma. We administered a questionnaire to identify characteristics of asthma, performed spirometric testing before and after administration of a bronchodilator, and measured exhaled NO levels in all participants. RESULTS: Exhaled NO was significantly correlated with the following markers of asthma disease control: asthma symptoms within the past 2 weeks (P =.02), dyspnea score (P =. 02), daily use of rescue medications (P =.01), and reversibility of airflow obstruction (P =.02). Exhaled NO levels were not correlated with the following markers of asthma disease severity: history of respiratory failure (P =.20), health care use (P =.08), fixed airflow obstruction (P =.91), or a validated asthma severity score (P =.19). Markers with relevance to both disease control and severity showed either a weak correlation (FEV(1) and FEV(1) percent predicted) or no correlation (controller drug use) with exhaled NO. CONCLUSION: We conclude that exhaled NO levels are correlated predominantly with markers of asthma control rather than asthma severity. Monitoring of exhaled NO may be useful in outpatient asthma management. PMID- 11031335 TI - Risk factors associated with glucocorticoid-induced adverse effects in children with severe asthma. AB - BACKGROUND: Although high-dose inhaled glucocorticoids (GCs) with or without chronically administered oral GCs are often used in children with severe persistent asthma, the adverse effects associated with their use have not been well-described in this patient population. OBJECTIVE: We sought to determine the GC-induced adverse effects profile of older children with severe persistent asthma. METHODS: A chart review of 163 consecutive children 9 years of age or older admitted to National Jewish for difficult to control asthma was done. RESULTS: The population studied consisted mostly of adolescents (mean +/- SD age, 14.4 +/- 2.1 years) with severe asthma receiving high-dose inhaled GC therapy (1675 +/- 94 microg/d) and averaging 6 systemic GC bursts per year. 50% required chronic oral GC therapy. GC-associated adverse effects were common and included hypertension (88%), cushingoid features (66%), adrenal suppression (56%), myopathy (50%), osteopenia (46%), growth suppression (39%), obesity and hypercholesterolemia (30%), and cataracts (14%). Height standard deviation scores of -0.44, -1.22, and -0.93 for those receiving intermittent, alternate day, and daily oral GCs, respectively, were smaller (less suppressed) than published values from the same institution before inhaled GC therapy (standard deviation scores of -1.26, -1.91, and -1.95, respectively). Osteopenia was strongly associated with growth suppression (odds ratio, 5.6; confidence interval, 2.7 11.8; P <.0001) and was found to be more common in female than male subjects, even after correcting for short stature (42% vs 18%, P <.006). CONCLUSIONS: GC associated adverse effects are still unacceptably common among children with severe asthma, even in those not receiving chronically administered oral GC therapy yet receiving high-dose inhaled GCs. Therefore close monitoring and proper intervention are warranted, especially in female subjects, who appear to be at greater risk for osteopenia. There is clearly a need to consider alternative therapy or earlier intervention. The magnitude of growth suppression, while still a problem, appeared to be less severe with the addition of inhaled GC therapy. This observation suggests that high-dose inhaled GC therapy, by affording better asthma control and allowing less use of systemic therapy, has attenuated the growth-suppressive effects of poorly controlled asthma. PMID- 11031337 TI - Demonstration of a partially cryptic epitope of the major cat allergen Fel d 1: consequences for mAb-based standardization of cat extracts. AB - BACKGROUND: Antiallergen mAbs that do not recognize clinically important isoforms have been described, raising the question of the selection of mAbs for quantifying major allergens in order to standardize allergenic extracts. This question is even more critical if mAbs can discriminate between different forms of allergen molecules with the same amino acid sequence. OBJECTIVE: We sought to demonstrate that an anti-Fel d 1 mAb was able to discriminate between two forms of the major cat allergen independently of its amino acid sequence and to determine the relative importance and stability of both forms in various cat extracts. METHODS: Anti-Fel d 1 mAbs were raised in mice and characterized. By using two of these mAbs, a two-site ELISA was developed to quantify Fel d 1 in mass units. RESULTS: One of the anti-Fel d 1 mAbs developed was shown to specifically recognize a particular form of Fel d 1. A two-site ELISA with this mAb to capture Fel d 1 was able to quantify the allergen specifically in this form. It was then shown that (1) the quantitative importance of this form of Fel d 1 could vary from one cat extract to another, (2) Fel d 1 was converted into this form under certain conditions, and (3) both converted and unconverted forms of Fel d 1 may bear IgE epitopes that are specific. CONCLUSION: Although the present study emphasizes the issue of selecting mAbs that are not too specific to standardize allergenic extracts, it also demonstrates that very specific mAbs can be of interest, especially to verify the stability of allergens in extracts, since this stability might have clinical implications. PMID- 11031336 TI - The chinese herbal medicine formula MSSM-002 suppresses allergic airway hyperreactivity and modulates TH1/TH2 responses in a murine model of allergic asthma. AB - BACKGROUND: Asthma is a major public health problem worldwide, and the morbidity and mortality of asthma have increased in the past two decades. The reputed efficacy, low cost, and relative absence of side effects of traditional Chinese medicines (TCMs) have led to increasing interest in the use of TCMs for the treatment of asthma in Western countries. However, there are few well-controlled scientific studies on the efficacy, safety, and mechanisms of action of TCMs used to treat asthma. OBJECTIVE: The goal of this study was to investigate the effects of the Chinese herbal medicine formula MSSM-002, derived from TCMs used to treat allergic asthma, on a well-characterized mouse model of allergic asthma. METHODS: Mice sensitized intraperitoneally and challenged intratracheally with conalbumin were treated with MSSM-002 24 hours after the first intratracheal challenge. Dexamethasone-treated, saline solution sham-treated, and naive mice served as controls. The effects of MSSM-002 on allergic airway hyperreactivity, inflammation, antigen-specific antibody production, lung histologic features, and cytokine profiles were evaluated. RESULTS: MSSM-002 treatment virtually eliminated airway hyperreactivity and markedly reduced the total number of cells and the percent eosinophils in bronchoalveolar lavage fluids compared with the sham-treated group. Lung histologic features showed that MSSM-002 reduced inflammation and mucus production. These effects were equivalent to the effects of dexamethasone, but in contrast to the overall immunosuppressive effects of dexamethasone MSSM-002 treatment decreased antigen-specific IgE, IL-4, IL-5, and IL-13 levels without suppressing IgG2a and IFN-gamma synthesis. CONCLUSION: MSSM 002 exhibits anti-airway hyperresponsiveness, anti-airway inflammation, and immunoregulatory effects on T(H)1/T(H)2 responses, which may be useful for treatment of allergic asthma. PMID- 11031338 TI - Basophil and eosinophil accumulation and mast cell degranulation in the nasal mucosa of patients with hay fever after local allergen provocation. AB - BACKGROUND: Basophils and mast cells have certain similarities and are believed to be important in upper and lower respiratory allergy. OBJECTIVE: We sought to apply immunohistology to investigate the distribution and numbers of mast cells and basophils in the nasal mucosa after allergen provocation. METHODS: Allergen challenge with grass pollen was performed in 9 patients with seasonal allergic rhinitis out of the hay fever season. Nasal biopsy specimens were taken before and approximately 1 hour, 24 hours, and 1 week after intranasal allergen provocation. We determined relative numbers and their phenotypic characteristics by using mAbs specific for tryptase, chymase, IgE, eosinophils (BMK-13), and a new mAb against basophils (BB1) by using immunohistochemistry in frozen sections. RESULTS: In the nasal mucosa at baseline, practically no basophils were found in the epithelium. A significant increase in numbers was found in the epithelium and lamina propria of the nasal mucosa in the early phase as early as 1 hour after allergen provocation. At 24 hours and 1 week after allergen provocation, a significant increase in basophil numbers was found in the lamina propria only. The proportion of mast cells staining for chymase in the lamina propria decreased from a median of 38% (range, 0%-82%) to 14% (range, 0%-78%) within 1 hour of allergen provocation. The proportion of mast cells staining for chymase increased from 1% (range, 0%-86%) at baseline to 21% (range, 3%-85%) within 1 hour of allergen provocation. One week after provocation, mast cells returned to baseline numbers. A definite tissue eosinophilia was observed after allergen provocation. CONCLUSION: Basophil numbers are increased in the epithelium and lamina propria of the nasal mucosa of subjects with rhinitis after allergen challenge, with influx already apparent at 1 hour. Moreover, changes in mast cell percentages and numbers were observed within 1 hour of allergen provocation. PMID- 11031339 TI - Cytokine profile in minor salivary glands from patients with bronchial asthma. AB - BACKGROUND: T lymphocytes are important components of the bronchial inflammatory cell infiltrate in asthma. Because lymphocytes activated in the respiratory tract recirculate to remote glandular and mucosal sites, we previously studied the histologic features of minor salivary glands (MSGs) in bronchial asthma and found an airway-like inflammation with T-lymphocyte infiltration, the presence of mast cells that were often degranulated, and basement membrane thickening but no eosinophil infiltration. OBJECTIVE: We sought to investigate the cellular infiltration and cytokine profile in MSGs from untreated asthmatic subjects, steroid-treated asthmatic subjects, and control subjects and to compare these values with those found in bronchial biopsy specimens. METHODS: The cellular infiltration was studied by using immunohistochemistry. Cytokine messenger (m)RNA expression for IL-4, IL-5, and IFN-gamma was determined by using in situ hybridization and cytokine immunoreactivity with immunohistochemistry. RESULTS: A significant increase in CD4 and IL-4 mRNA(+) cells was observed in MSGs from asthmatic patients (both untreated and steroid-treated subjects) when compared with control subjects, which correlated with the clinical severity of asthma (FEV(1) and Aas score). In contrast to the bronchi, no IL-5 mRNA expression was observed in MSGs, and no difference was observed for MSG IFN-gamma mRNA between the groups. At the level of MSG protein expression, the 3 cytokines were seen, with a significant increase in IL-4 protein expression in steroid-treated asthmatic subjects compared with untreated asthmatic subjects and control subjects, but there were no differences between the groups in IL-5 and IFN-gamma protein expression. CONCLUSION: The cytokine mRNA expression pattern observed in the MSGs of asthmatic subjects was different from that found in the bronchi, suggesting a different local immune regulation. PMID- 11031340 TI - Macrophage subpopulations and macrophage-derived cytokines in sputum of atopic and nonatopic asthmatic subjects and atopic and normal control subjects. AB - BACKGROUND: Previous studies have shown a prominent macrophage signal in the bronchial mucosa from nonatopic (intrinsic) compared with atopic (extrinsic) asthmatic subjects. This observation might have represented an expansion of a proinflammatory macrophage population or a homeostatic mechanism to decrease T(H)2-type inflammation. OBJECTIVE: The aim of the study was to investigate the numbers of macrophages and macrophage subpopulations and the expression of IL-10 and IL-12 in sputum from asthmatic and control subjects. METHODS: Eight atopic asthmatic (AA) subjects, 10 nonatopic asthmatic (NAA) subjects, 6 atopic control (AC) subjects, and 7 normal control (NC) subjects underwent sputum induction. Macrophages were enumerated by using Romanowsky stain and immunocytochemistry (CD68). RFD1 (interdigitating cell marker) and RFD7 (mature phagocyte marker) mAbs were used for immunocytochemical phenotyping, whereas IL-10 and IL-12 messenger (m)RNA was examined with in situ hybridization by using (35)S-labeled riboprobes. The phenotype of cells expressing IL-10 or IL-12 mRNA was examined by simultaneous in situ hybridization and immunostaining. RESULTS: No differences in the numbers of CD68(+) macrophages and RFD1(+), RFD7(+), and RFD1(+)/RFD7(+) subpopulations were found between AA, NAA, AC, and NC subjects. However, the numbers of IL-10 and IL-12 mRNA(+) cells were increased in AA subjects compared with NAA, AC, and NC subjects (P <.05). No other differences were found among the groups. Most of the IL-10 and IL-12 mRNA(+) cells in sputum from asthmatic subjects were macrophages (>80%), with less than 10% of mRNA colocalizing to epithelial cells. CONCLUSIONS: Sputum macrophage numbers, unlike tissue macrophages, as previously reported, were not elevated in NAA subjects. Increased IL-10 and IL-12 expression in atopic asthma may indicate the existence of a homeostatic mechanism to decrease lung inflammation. The lack of such cytokines in intrinsic asthma may predispose to bronchial inflammation in these subjects. PMID- 11031341 TI - Relation of epidermal growth factor receptor expression to goblet cell hyperplasia in nasal polyps. AB - BACKGROUND: Because the epidermal growth factor receptor (EGFR) system regulates mucin production in airway epithelium, we hypothesized a role for this system in mucus hypersecretion that occurs in nasal polyposis. OBJECTIVE: We examined the relationship between goblet cell hyperplasia, EGFR expression, and inflammatory mediators produced by eosinophils and neutrophils in nasal polyp tissues. METHODS: Nasal polyp tissue samples from 8 patients and nasal turbinate biopsy specimens from 6 normal control subjects were examined for alcian blue/PAS staining, mucin MUC5AC (MUC5AC), and EGFR immunoreactivity and EGFR gene expression (in situ hybridization). We also examined the role of eosinophils and neutrophils in goblet cell hyperplasia. RESULTS: In control nasal mucosa alcian blue/periodic acid-Schiff- and MUC5AC-stained areas were 18.40% +/- 1.31% and 21.89% +/- 1.43%, respectively. In polyps the alcian blue/periodic acid-Schiff- and MUC5AC-stained areas were 51.30% +/- 5.85% and 52.07% +/- 6.58%, which was significantly larger than that found in control subjects (each comparison, P <.01). Four of 6 control specimens expressed EGFR messenger RNA and protein weakly in the epithelium. In polyps 4 of 8 specimens expressed EGFR gene and EGFR protein strongly; the EGFR-stained area was greater in hyperplastic than in pseudostratified epithelium. TNF-alpha immunoreactivity, expressed in eosinophils, was increased in EGFR-positive polyps compared with EGFR-negative polyps, suggesting a role for TNF-alpha in EGFR expression. Neutrophils were increased in the epithelium of EGFR-positive compared with EGFR-negative polyps, suggesting a role for these cells in mucin expression and in goblet cell degranulation. CONCLUSION: These data suggest a role for EGFR cascade in the regulation of goblet cell mucins in nasal polyps. Proof of concept will require clinical studies using selective EGFR inhibitors. PMID- 11031342 TI - Substance P and its receptor neurokinin 1 expression in asthmatic airways. AB - BACKGROUND: Neural mechanisms have been suggested to contribute to the pathogenesis of chronic asthma. The expression of neuropeptides such as substance P may be regulated by infectious pathogens, including Mycoplasma species. In contrast to substance P, the substance P receptor neurokinin 1 has not been examined at the protein level in asthmatic airways. OBJECTIVE: This study evaluated substance P and neurokinin 1 protein expression and mucus content in endobronchial biopsy specimens from normal control subjects and asthmatic subjects. Detection of Mycoplasma pneumoniae was performed in both biopsy and bronchoalveolar lavage specimens. METHODS: Biopsy specimens were collected from 10 normal control subjects and 18 asthmatic subjects before and after a 6-week treatment with a macrolide antibiotic (n = 11) or placebo (n = 7) and were stained for substance P, neurokinin 1, and mucus. M pneumoniae was evaluated by PCR. RESULTS: At baseline, compared with normal control subjects, asthmatic subjects demonstrated increased expression of substance P and neurokinin 1 and mucus content in the airway epithelium. Epithelial mucus content correlated with epithelial substance P expression (r (s) = 0.45, P =.04) and FEV(1) percent predicted (r (s) = -0.51, P =.019). After antibiotic treatment, both epithelial substance P and neurokinin 1 expression were significantly reduced in asthmatic subjects. M pneumoniae was found in 8 of 18 asthmatic subjects. Asthmatic subjects with M pneumoniae, compared with those without M pneumoniae, showed higher baseline epithelial neurokinin 1 expression, which was significantly reduced after antibiotic treatment (P =.02). CONCLUSION: Our data suggest that abnormalities in neural mechanisms may exist in the epithelium of asthmatic airways, and M pneumoniae is possibly involved in this process. Antibiotic intervention may be effective in the treatment of asthma partly through the downregulation of the neurogenic process. PMID- 11031343 TI - Evidence for the involvement of two different MHC class II regions in susceptibility or protection in allergic bronchopulmonary aspergillosis. AB - BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA) is a disease with uncertain pathology. Studies have suggested a pathogenic role for T(H)2 cells. Previously, we demonstrated, in a small group of patients, that T(H)2 reactivity to a major Aspergillus fumigatus antigen was restricted by HLA-DR2 or HLA-DR5 alleles. OBJECTIVES: We sought to confirm whether susceptibility to ABPA is exclusively associated with HLA-DR locus and to investigate the involvement of HLA-DQ genes in the development of ABPA. METHODS: Genomic DNA was extracted from patients with ABPA, patients without ABPA but with positive A fumigatus skin test responses and asthma or cystic fibrosis, and healthy control subjects. HLA-DR and HLA-DQ genes were detected by using low-resolution typing; high-resolution typing was done only on HLA-DR2- and HLA-DR5-positive individuals by using sequence specific primers (PCR-SSP). RESULTS: A significantly higher frequency of HLA-DR2 was observed in patients with ABPA versus those without ABPA (corrected P <.01) or healthy control subjects (corrected P <.01). Genotype analysis revealed that susceptibility to ABPA is associated with HLA-DR2 alleles DRB1*1503 and DRB1*1501 and, to a lesser extent, with the HLA-DR5 allele DRB1*1104. The presence of DR4 or DR7 alleles in non-DR2/5 patients with ABPA suggests that these alleles may also be contributing factors in this disease. Another striking observation was the significantly high frequency of HLA-DQ2 in patients without ABPA (67. 4%) compared with patients with ABPA (20.5%) and normal control subjects (37.7%), suggesting that these alleles may confer protection in the population without ABPA. CONCLUSION: These genetic studies suggest that HLA-DR molecules DR2, DR5, and possibly DR4 or DR7 contribute to susceptibility while HLA-DQ2 contributes to resistance and that a combination of these genetic elements determines the outcome of ABPA in patients with cystic fibrosis and asthma. PMID- 11031344 TI - Elevated chemokine levels in bronchoalveolar lavage fluid of patients with eosinophilic pneumonia. AB - BACKGROUND: Allergic lung inflammation is caused by accumulation and activation of different leukocyte subsets, such as eosinophils and T lymphocytes, in the lung. The chemokines are a large group of chemotactic cytokines that regulate leukocyte trafficking and may play an important role in allergic lung inflammation. OBJECTIVE: The purpose of this study was to evaluate the role of various chemokines, including eotaxin, RANTES, monocyte chemotactic protein (MCP) 1, macrophage inflammatory protein (MIP)-1beta, and IL-8 in the pathogenesis of eosinophilic pneumonia (EP). METHODS: The concentrations of eotaxin, RANTES, MCP 1, MIP-1beta, and IL-8 in bronchoalveolar lavage fluid (BALF) were measured by using ELISA in 15 patients with EP, 10 with idiopathic pulmonary fibrosis, 10 with sarcoidosis, and 11 healthy volunteers. RESULTS: Eotaxin in BALF was high only in patients with EP, and its level correlated significantly with the number of eosinophils in BALF of patients with EP and healthy volunteers. MCP-1 and MIP 1beta in BALF were preferentially increased in patients with EP. There was a significant correlation between MCP-1 levels and the number of macrophages in BALF of patients with EP and healthy volunteers. CONCLUSION: Our findings suggest that these CC chemokines contribute to the pathogenesis of EP through the specific recruitment of leukocyte subsets in the lung. PMID- 11031345 TI - Modulation of the atopy patch test reaction by topical corticosteroids and tar. AB - BACKGROUND: Pharmacologic studies in atopic eczema (AE) are difficult to standardize. Patients with AE differ in the stage of their skin disease (acute, subacute, chronic). OBJECTIVE: This study was designed to assess macroscopic and microscopic effects of pretreatment with topical glucocortico-steroids (GCSs) and tar on the atopy patch test (APT) reaction in patients with atopic eczema. METHODS: Nonlesional skin of the back of patients with AE (n = 6) was treated for 3 weeks at 3 different sites with triamcin-olonacetonide 0.1% in cetamacrogol ointment (GCSs), pix liquida 10% in cetamacrogol ointment (tar), and cetamacrogol ointment (vehicle), respectively. APTs were performed, and biopsy specimens were taken from all these sites (time = 0 and 24 hours) for immunohistochemical analysis. RESULTS: Treatment with both GCSs and tar was able to reduce the macroscopic outcome of the APT reaction. Furthermore, both treatment modalities had an almost equally inhibiting effect on the influx of T cells, eosinophils, and CD1(+), RFD1(+), IFN-gamma(+), and IL-4(+) cells, as well as on the percentage of vessels expressing the adhesion molecules vascular cell adhesion molecule 1 and E-selectin in response to epicutaneous aeroallergen challenge. CONCLUSION: Although both treatments significantly reduced the various cellular constituents of allergic inflammation, all cell types remained present. In addition, this study shows that the APT can be used to evaluate the effect of topical anti-inflammatory treatments on allergic inflammation in patients with AE. PMID- 11031346 TI - The maize major allergen, which is responsible for food-induced allergic reactions, is a lipid transfer protein. AB - BACKGROUND: Cereals are the most important nutritional component in the human diet. Food-induced allergic reactions to these substances therefore have serious implications, and exhaustive diagnosis is required. Such diagnosis is still difficult because of the incomplete knowledge about major cereal allergens. In particular, few food-induced allergic reactions to maize have been reported, and no information on the allergenic proteins is available. OBJECTIVE: Having observed several anaphylactic reactions to maize, we planned a study to identify maize major allergens and cross-reactivity with other cereals, as well as to peach because the majority of patients also reacted to Prunoideae fruits. METHODS: Twenty-two patients with systemic symptoms after maize ingestion and positive skin prick test responses and serum-specific IgE antibodies to maize were selected. The IgE-reactivity pattern was identified by SDS-PAGE and immunoblotting. The major allergen identified was then purified by HPLC and characterized by mass spectrometry, determination of the isoelectric point value, and N-terminal amino acid sequencing. RESULTS: Sera from 19 (86%) of the 22 patients recognized a 9-kd protein, thus confirming this as the maize major allergen. This protein had an isoelectric point of greater than 9, a molecular mass of 9047.0 d, and no glycosylation. Determination of its N-terminal sequence showed that it was a lipid transfer protein (LTP). By using immunoblotting inhibition experiments, we demonstrated that the LTP cross-reacts completely with rice and peach LTPs but not with wheat or barley LTPs. N-terminal sequence of the 16-kd allergen (recognized by 36% of patients) showed it to be the maize inhibitor of trypsin. This protein cross-reacts completely with grass, wheat, barley, and rice trypsin inhibitors. CONCLUSION: The major allergen of maize is an LTP with a molecular weight of 9 kd that is highly homologous with the peach LTP, the major allergen of the Prunoideae subfamily. PMID- 11031347 TI - Digestibility of allergens extracted from natural rubber latex and vegetable foods. AB - BACKGROUND: Several cross-reactive allergens are now known to be involved in the defense responses of higher plants. Such proteins are drawing the attention of plant breeders because of their antimicrobial or stress-alleviating activities. Plants genetically modified to express defense-related proteins are being developed. The current concern is focused on the allergenicity of these intentionally expressed proteins. OBJECTIVE: It is believed that food allergens are proteins resistant to digestion. Digestibility tests have been accepted as an appropriate method for evaluating the allergenicity of newly introduced proteins. In this study we investigated the usefulness of this method for detecting allergens from natural rubber latex and vegetable foods. METHODS: Proteins were extracted from rubber latex, potato, and 5 kinds of fruits. Simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) were used for the digestibility test. An aliquot of each digest was periodically withdrawn and analyzed. Allergens were detected with pooled sera from individuals with latex allergy or patients given a diagnosis of oral allergy syndrome. RESULTS: Most latex and vegetable food proteins were digested by the SGF within 4 minutes. Numerous allergens were also decomposed by the SGF within 8 minutes. Although vegetable food allergens were relatively stable in the SIF, kiwi allergens were substantially degraded by the SIF within 16 hours. CONCLUSION: The pronounced lability of the plant-derived allergens was thought to reflect the discrete sensitization and elicitation processes of patients with latex-fruit syndrome or oral allergy syndrome. These results indicate that the allergenicity of a newly expressed protein should be carefully evaluated according to not only its digestibility but also other important properties. PMID- 11031348 TI - The effects of roasting on the allergenic properties of peanut proteins. AB - BACKGROUND: Because of the widespread use of peanut products, peanut allergenicity is a major health concern in the United States. The effect or effects of thermal processing (roasting) on the allergenic properties of peanut proteins have rarely been addressed. OBJECTIVE: We sought to assess the biochemical effects of roasting on the allergenic properties of peanut proteins. METHODS: Competitive inhibition ELISA was used to compare the IgE-binding properties of roasted and raw peanut extracts. A well-characterized in vitro model was used to test whether the Maillard reaction contributes to the allergenic properties of peanut proteins. The allergic properties were measured by using ELISA, digestion by gastric secretions, and stability of the proteins to heat and degradation. RESULTS: Here we report that roasted peanuts from two different sources bound IgE from patients with peanut allergy at approximately 90 fold higher levels than the raw peanuts from the same peanut cultivars. The purified major allergens Ara h 1 and Ara h 2 were subjected to the Maillard reaction in vitro and compared with corresponding unreacted samples for allergenic properties. Ara h 1 and Ara h 2 bound higher levels of IgE and were more resistant to heat and digestion by gastrointestinal enzymes once they had undergone the Maillard reaction. CONCLUSIONS: The data presented here indicate that thermal processing may play an important role in enhancing the allergenic properties of peanuts and that the protein modifications made by the Maillard reaction contribute to this effect. PMID- 11031350 TI - Resolution of peanut allergy. PMID- 11031349 TI - Subjects with allergic reactions to drugs show in vivo polarized patterns of cytokine expression depending on the chronology of the clinical reaction. AB - BACKGROUND: The mechanisms involved in adverse drug reactions with an immunologic basis (ADRIB) can be antibody dependent, mainly IgE or T cell dependent (sensitized T cells). These mechanisms are regulated by a number of cytokines, including IL-2, IL-4, IL-5, IFN-gamma, and TNF-alpha, which follow the classical T(H)1/T(H)2 immunologic paradigm. Although evidence for this has been seen in ex vivo studies, the results are heterogeneous, and few in vivo studies have been carried out in subjects with ADRIB. OBJECTIVE: We studied a group of patients who experienced either immediate reactions (n = 10) or nonimmediate reactions (n = 9) to drugs to determine the cytokine pattern profile during the acute stage of the response, as well as after recovery. METHODS: PBMCs were taken at different time intervals of 24 hours or less and 7, 15, and 30 days after the onset of the reaction, and the specific cytokine transcription and production were determined by using quantitative competitive RT-PCR and ELISA, respectively. RESULTS: There was a transient polarized pattern corresponding to a T(H)1 response with IL-2, IFN-gamma, and TNF-alpha in nonimmediate reactions and to a T(H)2 response with IL-4 in immediate reactions. CONCLUSIONS: This is the first in vivo demonstration of these T(H)1/T(H)2 patterns in subjects with ADRIB and confirms that an immunologic process is occurring related to the mechanisms involved in the pathologic manifestation. These findings are relevant to the understanding of the pathophysiologic mechanisms involved in ADRIB, suggesting that further studies in this direction are warranted. PMID- 11031351 TI - Prohevein-like defense protein of tobacco is a cross-reactive allergen for latex allergic patients. PMID- 11031352 TI - Simultaneous occupational allergy to a carnation and its parasite in a greenhouse worker. PMID- 11031353 TI - Extrinsic allergic alveolitis caused by cabreuva wood dust. PMID- 11031354 TI - Association of decreases in asthma mortality with increases in sales of inhaled corticosteroids. PMID- 11031356 TI - The dose of intravenous hydrocortisone in acute wheezing and dyspnea. PMID- 11031359 TI - Ultrasonic coagulation in oslar disease: hereditary hemorrhagic telangiectasia. PMID- 11031360 TI - Lymphangioleiomyomatosis (LAM): a review of clinical and morphological features. AB - A review is presented of the clinical and morphological manifestations of lymphangioleiomyomatosis (LAM), a systemic disorder of unknown etiology that affects women. The clinical features include dyspnea, hemoptysis, recurrent pneumothorax, chylothorax, and chylous ascites. It is characterized by: 1) proliferation of abnormal smooth muscle cells (LAM cells) in pulmonary interstitium and along the axial lymphatics of the thorax and abdomen; 2) thin walled pulmonary cysts, and 3) a high incidence of angiomyolipomas. The pulmonary cystic lesions have a characteristic appearance on high resolution computed tomography. The most specific method for diagnosing LAM is lung biopsy to demonstrate the presence of LAM cells, either by their characteristic histological appearance or by specific immunostaining with HMB-45 antibody. LAM cells differ in several important respects from the types of smooth muscle cells normally present in lung. Their reactivity with HMB-45 antibody is localized in stage I and stage II melanosomes. LAM cells show additional evidence of incomplete melanogenesis, and the significance of these observations remains to be determined. Two types of LAM cells are recognized: 1) small, spindle-shaped cells that are centrally located in the LAM nodules and are highly immunoreactive for matrix metalloproteinase-2 (MMP-2), its activating enzyme (MT-1-MMP), and proliferating cell nuclear antigen (PCNA), and 2) large, epithelioid cells that are distributed along the periphery of the nodules and show a high degree of immunoreactivity with HMB-45 antibody and with antibodies against estrogen and progesterone receptors. Types of treatment used for LAM include oophorectomy, administration of Lupron or progesterone and in very severe cases, pulmonary transplantation (following the onset of respiratory insufficiency, not relieved by O(2)). PMID- 11031361 TI - Clinicopathological analysis of premature infants treated with artificial surfactant. AB - OBJECTIVE: Our aim was to obtain new information about the relationship between infant responses to surfactant replacement therapy and histopathological changes in vital organs. STUDY DESIGN: To accomplish this, the autopsy findings and clinical backgrounds of 41 very low birth weight infants (gestational week 25.6 +/- 2.3; birth weight 806.4 +/- 251.6g) who had died after receiving surfactant replacement therapy were reviewed, and those who responded to therapy were compared with those who did not. Responders were infants in whom the required FiO(2) declined by > 20% or mean airway pressure declined by > 20% within six hours of instilling surfactant (n=18); non-responders were infants who did not meet those criteria (n=23). RESULT: Gestational age, birth weight and time at treatment were similar in responders and non-responders, but survival was significantly longer in responders. The incidences of hyaline membrane disease, pulmonary interstitial emphysema, hemorrhagic necrosis and parenchymal degeneration of the liver and kidney were all higher in non-responders, whereas the incidences of bronchopulmonary dysplasia and pneumonia were higher in responders. Prior to treatment, acidosis and hypothermia were significantly more severe in non-responders, and perinatal complications, such as fetal distress and intrauterine infection, were observed more often in non-responders. Substantial degradation of vital organs had already occurred during the early post-natal or intrauterine life of the non-responders, which would be expected to interfere with the clinical response to instilled surfactant. CONCLUSION: It is anticipated that in the future improved monitoring of immature fetuses will be indispensable to improve intrauterine fetal management and to achieve better control over the timing and mode of delivery. PMID- 11031362 TI - Peripheral directional atherectomy evaluated with a rotational digital angiography system. AB - BACKGROUND: One of the factors that affect the recurrence rate after peripheral directional atherectomy (DA) is the degree of residual stenosis. A new method of peripheral DA to reduce residual stenoses was evaluated with a rotational digital angiography (RDA) system that provides both angiography and fluoroscopy at multiple projections within 360 degrees. PATIENTS AND METHODS: Between March 1995 and July 1999, severe short segmental stenoses of six iliac arteries and two superficial femoral arteries (SFA) in eight patients were treated with the Simpson DA catheter under RDA system guidance. After pre-procedural RDA evaluation, the first series of DA were performed under ordinary PA fluoroscopic guidance. The residual stenoses were evaluated with RDA. If the residual stenoses exceeded 30%, a second series of DA were performed covering the residual plaque with the cutter window of the DA catheter. To this purpose the fluoroscopy of the RDA system was fixed in the direction in which the residual stenoses were largest and most eccentric. The end point was defined to be a residual stenosis of less than 30% evaluated with the RDA system, and the procedures were repeated until the end point was achieved. RESULTS: Five of six iliac artery lesions were curved at the pre-procedural RDA evaluation. After the first series of DA, only two of six iliac lesions but all SFA lesions achieved the end point. Among the four other iliac lesions, three achieved the end point with one or two additional series of DA using the RDA system guidance to control the selective cuts of the residual plaques. One patient had a residual stenosis of 50% because the procedure could not be completed by balloon rupture of the DA catheter. In the patients with iliac stenoses, there was no final residual stenosis in one, and the range was from 20% to 25% in the four patients. The residual stenoses were located on the greater curvature side of the curved artery in three of these four patients. CONCLUSION: The RDA system is a valuable tool in aiding reduction of the residual stenoses during peripheral DA. Minimal stenoses often remain on the greater curvature side of the wall because the rigid and straight metallic capsule (cylindrical housing) of the Simpson DA catheter does not completely fit the curved wall. This phenomenon was thought to be a mechanical limitation of this device. PMID- 11031363 TI - Expression of endothelin-1 after endothelial denudation of thoracic aortas in experimental hypercholesterolemic rats. AB - Although endothelin-1 (ET-1) is involved in balloon-induced neointima formation, the role of ET-1 in balloon-induced neointima formation in hypercholesterolemia is unclear. In addition, it remains to be determined whether ET-1 is produced by endothelial cells or vascular smooth muscle cells, or both. We investigated tissue immunoreactive ET-1 levels by immunoblot analysis, localization of ET-1 immunoreactivity by immunohistochemistry, and expression of preproET-1 mRNA by in situ hybridization in balloon-induced neointima formation in experimental hypercholesterolemic rats. Serum total cholesterol levels were significantly higher (p< 0.01) in the 5%cholesterol-diet group (194 +/- 17 mg/dl, n=20) than in the normal-diet group (64 +/- 2 mg/dl, n=20). Before and after endothelial denudation, plasma ET-1 levels and tissue immunoreactive ET-1 levels were significantly higher in cholesterol-diet rats. The expression of preproET-1 mRNA by in situ hybridization was observed in the nuclei of endothelial cells, but not medial smooth muscle cells in normal- or cholesterol diet rats. After endothelial denudation, plasma ET-1 levels and serum total cholesterol levels did not change in either the normal- or the cholesterol-diet rats. Tissue level of ET-1 tended to increase at 3 days after denudation in normal-diet rats (1.0 +/- 0.1 vs 2.6 +/ 0. 2 density ratio, p< 0.05), although endothelial cells had not yet regenerated. The expression of preproET-1 mRNA by in situ hybridization was not observed at 3 days after endothelial denudation in either endothelial or medial smooth muscle cells in normal-diet rats. Four weeks after denudation, regeneration of endothelial cells was almost complete, and an intimal hyperplasia was observed. Tissue ET-1 levels were significantly elevated 4 weeks after endothelial denudation in normal-diet rats (1.0 +/- 0.1 vs 7.6 +/- 0.2 density ratio, p< 0.05). The expression of preproET-1 mRNA by in situ hybridization was observed in the nuclei of regenerated endothelial cells after endothelial denudation, and in smooth muscle cells migrating into the intima, but was not observed in medial smooth muscle cells in normal-diet rats. A similar pattern was observed in cholesterol-diet rats. We concluded that ET-1 was involved in neointima formation and that ET-1 was produced by both endothelial and neointimal smooth muscle cells, but not medial smooth muscle cells after endothelial denudation in experimental hypercholesterolemic rats. PMID- 11031365 TI - Recent findings on bacterial infection from both basical and clinical aspects PMID- 11031364 TI - Prediction of uterine dehiscence by measuring lower uterine segment thickness prior to the onset of labor: evaluation by transvaginal ultrasonography. AB - OBJECTIVE: Lower uterine segment thickness was measured by transvaginal ultrasound examination and its correlations with the occurrence of uterine dehiscence and rupture was examined. METHODS: The thickness of the muscular layer of the lower uterine segment was measured in 186 term gravidas with previous uterine scars and its correlation with uterine dehiscence/rupture was investigated. RESULTS: Uterine dehiscence was found in 9 cases or 4.7%. There were no cases of the uterine rupture. The thickness of the lower uterine segment among the gravidas with dehiscence was significantly less in than those without dehiscence (p< 0.01). The cut-off value for the thickness of the lower uterine segment was 1.6 mm as calculated by the receiver operating characteristic curve. The sensitivity was 77.8%; specificity 88.6%; positive predictive value 25.9%; negative predictive value 98.7%. CONCLUSION: Measurement of the lower uterine segment is useful in predicting the absence of dehiscence among gravidas with previous cesarean section. If the thickness of the lower uterine segment is more than 1.6 mm, the possibility of dehiscence during the subsequent trials of labor is very small. PMID- 11031366 TI - [Helicobacter pylori and stomach diseases:from clinical point of view]. PMID- 11031368 TI - [Topics of drug resistance bacteria and legionella pneumonia]. PMID- 11031367 TI - [Helicobacter pylori and gastric pathology]. PMID- 11031369 TI - [Tuberculosis: global overview and current issues in Japan]. PMID- 11031370 TI - [Flesh-eating bacteria: streptococcal toxic shock syndrome]. PMID- 11031371 TI - [A study of metallic mercury polluting a room after being spilled from a sphygmomanometer]. AB - Mercury spilled from a mercurial sphygmomanometer on a hot carpet can vaporize and pollute the environment. We observed the vaporization of mercury in model experiments. Mercury (0.15g) was heated on a hot carpet and the near-by air was sampled with a midget impinger. The evaporated mercury levels were 5.0, 6.3, 8.1 and 10.0mg/m(3) at 20, 40, 60 and 80 minutes, respectively at a height of 30cm from carpet. The result indicated that even if a small quantity of mercury remained on the hot carpet, it could evaporate and pollute the indoor air. Little is known about the influence on human health of low mercury exposure, especially on children. In order not to pollute the air, we need to pay attention to mercury. PMID- 11031372 TI - Successful removal of a stone and an expandable metallic stent from the biliary tract of a patient with acute occlusive pyogenic cholangitis. AB - We removed a biliary stone and the metallic stent placed two years previously in a patient with benign biliary strictures. An 80-year-old woman who had been inplanted with an expandable metallic stent (EMS) to prevent obstruction by a large common bile duct stone about two years before as an emergency measure in another hospital, was afficted with acute occlusive pyogenic cholangitis (AOPC) and hospitalized in our hospital. After treating the AOPC, we successfully removed the EMS with a cholangioscope and normal biopsy forceps through the percutaneous transhepatic channel under fluoroscopy. The type of the EMS was Accufulex stent. To remove it was easier than expected. Once it started to unravel, it was removed from the common bile duct within a few minutes. PMID- 11031374 TI - Intracranial lipomas: demonstration by computed tomography and magnetic resonance imaging. AB - We present two cases of a very rare tumor, intracranial lipoma, diagnosed by computed tomography (CT) and magnetic resonance imaging (MRI). In one case, the lipoma was in the superior cerebellar cistern, the other was in the periphery of the corpus callosum. In the case in which MRI was used, identification of the lipoma using a routine MRI examination was difficult. These cases are reported now because the incidental diagnosis of intracranial lipoma is likely to increase due to advanced neuroradiological techniques such as CT and MRI. PMID- 11031373 TI - [The first case of living-related liver transplantation in Nippon Medical School Hospital]. PMID- 11031375 TI - Granulocytic sarcoma of the jejunum: pit hole in immunohistochemical approach. PMID- 11031376 TI - Standards for the evaluation of hospital infection control policies and procedures. PMID- 11031377 TI - A nonhealing ulcer on the face. PMID- 11031378 TI - Proxy measures and human immunodeficiency virus care. PMID- 11031379 TI - Human immunodeficiency virus infection in rural practice. PMID- 11031381 TI - Targeted advertising in medical journals PMID- 11031380 TI - Targeted advertising in medical journals. PMID- 11031382 TI - Dextromethorphan abuse among youth. PMID- 11031383 TI - Pressure, heat, and cold help relieve headache pain. PMID- 11031384 TI - The urgent need to improve hypertension care. AB - Hypertension is the most prevalent health problem among adult primary care patients, but its recognition and treatment are suboptimal. Although there is ample evidence from several large-scale randomized, controlled studies that treatment of hypertension reduces morbidity and mortality, current management of hypertension is characterized by underdiagnosis, misdiagnosis, undertreatment, overtreatment, and misuse of medications. As a result, roughly 75% of the estimated 50 million adults with hypertension in the United States are at increased risk for vascular complications. Optimal therapy requires careful attention to patients' age, sex, race, diet, exercise, tobacco use, comorbid conditions, choice of antihypertensive drug treatment, compliance with treatment, and achievement of blood pressure control. Other issues that deserve scrutiny are accuracy of the initial diagnosis, self-monitoring of blood pressure, and the advisability of attempting reduction of dosage or possible withdrawal from administration of antihypertensive drug treatment in patients whose blood pressures have been controlled for 1 year or more. Physicians' knowledge and use of the Sixth Joint National Committee on Prevention, Detection, and Treatment of High Blood Pressure report are deficient. Several responses to this current crisis in care of hypertensive patients are reviewed, including computer-aided management, medical chart audit, academic detailing, and a nurse case manager using prepared algorithms in consultation with the physician. PMID- 11031385 TI - Traumatic life events in primary care patients: a study in an ethnically diverse sample. AB - OBJECTIVE: To examine among immigrants and others seeking primary care: (1) the prevalence, types, and predictors of traumatic life events; and (2) the relations among traumatic life events, psychiatric disorders, and utilization of primary care services. DESIGN: Survey with structured diagnostic interview. SETTING: Community-based, university-affiliated primary care clinic in southern California. PARTICIPANTS: Fourteen hundred fifty-six adult patients representing 4 ethnic groups (Mexican immigrants, Central American immigrants, US-born Latinos of Mexican descent, and US-born non-Latino whites). DEPENDENT MEASURES: Rates of traumatic events measured with the Posttraumatic Stress Disorder section of the Diagnostic Interview Schedule; psychiatric disorders identified by the Composite International Diagnostic Interview using Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria; physical functioning (Short Form Health Survey); and the number of medical clinic visits during a 6-month period. RESULTS: Nearly 10% of patients had experienced a traumatic event in the previous year, and 57% had experienced at least 1 during their lifetimes. The most common forms of trauma were interpersonal violence occurring outside the family (21%), acute losses or accidents (17%), witnessing death or violence (13%), and domestic violence (12%). When compared with the US-born non-Latino whites, Mexican immigrants were half as likely, and Central American immigrants were 76% more likely, to report having experienced a traumatic event. Married individuals were significantly less likely to report traumas. Traumatic experiences, female gender, and non-Latino ethnicity were associated with the presence of a psychiatric disorder. One-year and lifetime psychiatric disorders were associated with poorer physical functioning and an increased number of clinic visits during a 6-month period. CONCLUSIONS: Traumatic life events are common and associated with psychiatric disorders other than posttraumatic stress disorder in an ethnically diverse sample of primary care patients. Psychiatric disorders, in turn, are strongly associated with poor physical functioning and higher rates of primary care utilization. Screening for traumatic experiences should accompany assessments of psychiatric disorders to ensure adequate treatment of patients seeking primary care services. PMID- 11031386 TI - Social, cultural, and clinical dimensions of traumatic experiences of primary care patients. PMID- 11031387 TI - Detecting symptoms of alcohol abuse in primary care settings. AB - BACKGROUND: Studies on alcohol abuse are frequently based on patients who meet minimum diagnostic criteria, thus ignoring patients with individual symptoms of harmful or hazardous use. Consequently, we are unable to characterize alcohol abusing patients with sufficient clarity to effectively focus screening for primary prevention. OBJECTIVE: To determine the prevalence of harmful and hazardous use of alcohol, assess screening instruments for detecting alcohol abuse or dependence, and assess the impact of alcohol use on other diagnoses treated in outpatient settings. DESIGN: Survey (cross-sectional study). SETTING: Hospital-based outpatient clinic. PARTICIPANTS: Three hundred randomly selected adults (aged 18 years and older). MAIN OUTCOME MEASURE: Diagnosis of alcohol abuse or dependence based on the Diagnostic Interview Schedule (DIS). RESULTS: About 18% met DSM-IV (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) criteria of abuse or dependence while almost 50% had at least one symptom of harmful or hazardous use. The T-ACE questionnaire, a modified version of the CAGE questionnaire, was the most effective screening instrument for both men and women. Selected diagnoses, personal characteristics such as family history of alcohol abuse, and self-reported patterns of alcohol use could identify patients likely to meet diagnostic criteria. CONCLUSIONS: Many symptoms of substance use disorders are not adequately addressed in outpatient practice. Little is known about how alcohol use in varying quantities affects health care utilization and treatment of conditions commonly seen in outpatient medicine. Consequently, we lack a full appreciation of the burden of disease borne by alcohol use and have yet to achieve a universally accepted method of approaching primary and secondary prevention of alcohol-related problems. PMID- 11031388 TI - A questionnaire survey of family practice physicians' perceptions of bereavement care. AB - Survey responses were obtained from 113 family physicians in an exploratory investigation of bereavement care by family physicians. Respondents generally indicated that they believed that bereavement presented significant health risks to their patients and that the identification and treatment of bereaved patients was an important part of their role. However, physicians were highly variable in how they reported identifying and responding to bereaved patients in terms of counseling, addressing spiritual concerns, and medically treating symptoms. The "grief-responsive" physician is described based on these data. We document the interest and need for training in bereavement care. PMID- 11031389 TI - Good grief. PMID- 11031390 TI - Attitudes of Israeli family physicians toward clinical guidelines. AB - BACKGROUND: Many clinical guidelines (CGs) have been written during the past few years. Although family physicians (FPs) stand to benefit from many of these CGs, incorporating new CGs into daily practice seems to present a challenge. OBJECTIVE: To evaluate Israeli FPs' attitudes toward CGs. METHODS: We administered an anonymous questionnaire to general practitioners, residents, and board-certified FPs who participated in continuing medical education programs throughout Israel during May and June 1998. Our survey focused on physician attitudes and behaviors regarding CGs in general, and to CGs for treating the patient with diabetes mellitus (DM) in particular. The CGs for patients with DM have recently been promoted in the context of primary care quality improvement programs. Respondents also provided demographic and professional data. RESULTS: Of the 404 questionnaires distributed, 293 questionnaires were returned for a response rate of 83%. The average (+/-SD) age of respondents was 40.2 +/- 7.0 years, with a mean (+/-SD) of 13.3 +/- 8.0 years in practice. Overall, opinion regarding CGs was positive. About half of the respondents thought CGs improved patient compliance. Comparisons between the various physician groups highlighted several notable differences. Two thirds of the general practitioners believed CGs improved patient compliance, while this was true of only one third of the FPs (P<.001). Most FPs (62%) and senior residents (69%) felt CGs did not constrain their clinical freedom, while less than half of the general practitioners and junior residents felt this way (P =.045). Eighty-three percent of all respondents agreed that the CGs for the treatment of DM were able to be implemented, and 75% believed the CGs assisted them in the management of patients with DM. Whereas 39% expressed concern about being able to adapt generic CGs to individual patient needs, only 27% (P =.002) felt this way about the DM CGs. The vast majority (92%) were interested in understanding the scientific evidence supporting CGs as a prerequisite to adopting them. Most respondents preferred limiting CG length to a maximum of 5 pages. CONCLUSIONS: We found support among Israeli FPs for the use of CGs. Clinical guidelines seem to be used in the field, in particular those developed for treating DM. In light of our findings, attention should be focused on optimally tailoring new CGs to meet scientific standards and crafting them to suit the preferences of local FPs. PMID- 11031391 TI - Sexual orientation and health: comparisons in the women's health initiative sample. AB - CONTEXT: Little is known about older lesbian and bisexual women. Existing research rarely compares characteristics of these women with comparable heterosexual women. OBJECTIVE: To compare heterosexual and nonheterosexual women 50 to 79 years on specific demographic characteristics, psychosocial risk factors, screening practices, and other health-related behaviors associated with increased risk for developing particular diseases or disease outcomes. DESIGN: Analysis of data from 93,311 participants in the Women's Health Initiative (WHI) study of health in postmenopausal women, comparing characteristics of 5 groups: heterosexuals, bisexuals, lifetime lesbians, adult lesbians, and those who never had sex as an adult. SETTING: Subjects were recruited at 40 WHI study centers nationwide representing a range of geographic and ethnic diversity. PARTICIPANTS: Postmenopausal women aged 50 to 79 years who met WHI eligibility criteria, signed an informed consent to participate in the WHI clinical trial(s) or observational study, and responded to the baseline questions on sexual orientation. MAIN OUTCOME MEASURES: Demographic characteristics, psychosocial risk factors, recency of screening tests, and other health-related behaviors as assessed on the WHI baseline questionnaire. RESULTS: Although of higher socioeconomic status than the heterosexuals, the lesbian and bisexual women more often used alcohol and cigarettes, exhibited other risk factors for reproductive cancers and cardiovascular disease, and scored lower on measures of mental health and social support. Notable is the 35% of lesbians and 81% of bisexual women who have been pregnant. Women reporting that they never had sex as an adult had lower rates of Papanicolaou screening and hormone replacement therapy use than other groups. CONCLUSIONS: This sample of older lesbian and bisexual women from WHI shows many of the same health behaviors, demographic, and psychosocial risk factors reported in the literature for their younger counterparts, despite their higher socioeconomic status and access to health care. The lower rates of recommended screening services and higher prevalence of obesity, smoking, alcohol use, and lower intake of fruit and vegetables among these women compared with heterosexual women indicate unmet needs that require effective interactions between care providers and nonheterosexual women. PMID- 11031392 TI - Obese women's perceptions of their physicians' weight management attitudes and practices. AB - BACKGROUND: Obesity has reached epidemic proportions in the United States. Primary care physicians will see increasing numbers of patients with long-term weight management problems. OBJECTIVE: To examine obese women's perceptions of their physicians' weight management attitudes and practices. DESIGN AND SETTING: Women who participated in obesity trials at a university clinic completed a questionnaire that assessed their views of weight control provided by their primary care physician. PARTICIPANTS: The patients were 259 women whose age was 44.0 +/- 10.0 years; weight, 96.7 +/- 13.2 kg; and body mass index (calculated as weight in kilograms divided by the square of height in meters), 35.2 +/- 4.5 (all data given as mean +/- SD). MAIN OUTCOME MEASURES: Using 7-point scales (1 indicates low; and 7, high), patients rated their satisfaction with care provided for their general health and that for their obesity. They also identified methods their physician recommended for weight management and the frequency of negative interactions with their physician concerning weight control. RESULTS: Participants were generally satisfied with the care they received for their general health and with their physicians' medical expertise (mean scores, 6. 1 and 6.2, respectively). They were significantly (P<.001) less satisfied with care for their obesity and with their physicians' expertise in this area (mean scores, 4.1 and 4.3, respectively). Almost 50% reported that their physician had not recommended any of 10 common weight loss methods, and 75% indicated they looked to their physician a "slight amount" or "not at all" for help with weight control. Only a small minority of patients (0.4%-8.0%) reported frequent, negative interactions with physicians concerning their weight. CONCLUSIONS: The last finding helps allay concerns that obese patients are routinely treated disrespectfully by physicians when discussing weight. The challenge, however, for primary care physicians appears to be providing patients better assistance with weight management. PMID- 11031393 TI - Antiviral therapy for herpes zoster: randomized, controlled clinical trial of valacyclovir and famciclovir therapy in immunocompetent patients 50 years and older. AB - OBJECTIVE: To compare the efficacy and safety of valacyclovir hydrochloride and famciclovir for the treatment of herpes zoster. DESIGN: A double-blind, randomized, controlled, multicenter clinical trial in which patients received 7 days of treatment and were followed up for 24 weeks. SETTINGS: Patients reported directly to specialist centers or were referred from primary care centers. PATIENTS: There were 597 otherwise healthy immunocompetent outpatients, aged 50 years and older, who presented within 72 hours of onset of zoster rash. INTERVENTIONS: Treatment with valacyclovir hydrochloride (1 g 3 times daily) or famciclovir (500 mg 3 times daily) for 7 days. MAIN OUTCOME MEASURES: Resolution of zoster-associated pain and postherpetic neuralgia, rash healing, and treatment safety. RESULTS: Intent-to-treat analysis did not detect statistically significant differences for valacyclovir vs famciclovir on resolution of zoster associated pain (hazard ratio, 1. 02; 95% confidence interval, 0.84-1.23; P =.84). Furthermore, no differences were evident between treatments on rash healing rates and on a range of analyses of postherpetic neuralgia. Safety profiles for valacyclovir and famciclovir were similar, with headache and nausea being the more common adverse events. CONCLUSIONS: Valacyclovir treatment is comparable to famciclovir treatment in speeding the resolution of zoster associated pain and postherpetic neuralgia. Current wholesale prices indicate that valacyclovir is the more cost-effective treatment for herpes zoster ($83.90 vs $140.70 per course). PMID- 11031394 TI - Predictors of screening mammography: implications for office practice. AB - BACKGROUND: The completion of annual screening mammography and other preventive health services among women aged 50 years and older remains an important quality of care indicator. METHODS: A biracial sample of 843 rural women (aged > or =50 years) from a population-based sample reported demographic and preventive health services utilization in the last year including the completion of screening mammography. Bivariate analysis and logistic regression were used to investigate the extent to which completion of other screening examinations, including Papanicolaou (Pap) smears and clinical breast examination, is associated with successful completion of mammography relative to demographic and health service variables. RESULTS: The completion of mammography was associated with age, race, education, health insurance, and the presence of a regular primary care physician, but the strongest predictors were the completion of a clinical breast examination and/or a Pap smear. CONCLUSIONS: Women who receive a clinical breast examination and/or a Pap smear appear far more likely to receive screening mammography, suggesting a synergy in screening services. The relative efficacy of interventions to increase the completion of clinical breast examinations as well as other age-appropriate preventive services during routine office visits or during a single preventive services office visit should be further explored in primary care settings. Residency programs should provide training on the successful incorporation of such services into office practice patterns in an effort to continually improve quality of care. PMID- 11031395 TI - Prevalence of anxiety, depression, and substance use disorders in an urban general medicine practice. AB - BACKGROUND: Most research on the prevalence of mental disorders in primary care has been conducted in practices that serve middle- and upper-income patients. OBJECTIVE: To determine the prevalence of major mental disorders in a primary care practice that serves a predominantly low-income immigrant patient population. DESIGN: Cross-sectional survey; criterion standard. SETTING: Urban general medicine practice. PARTICIPANTS: Systematic sample of consecutive adult patients with scheduled appointments. Of 1266 approached eligible patients, 1007 (80%) participated. MAIN OUTCOME MEASURES: PRIME-MD Patient Health Questionnaire major depression, generalized anxiety disorder, panic disorder, alcohol use disorder, and suicidal ideation; drug use disorder; functional status; work loss; family distress; and mental health treatment. RESULTS: Major depression (18. 9%), generalized anxiety (14.8%), panic (8.3%), and substance use (7. 9%) disorders and suicidal ideation (7.1%) were highly prevalent. Many patients had more than 1 disorder (range, 36.3% [substance use disorder] to 76.9% [panic disorder]). In multivariate analyses, each disorder was significantly associated with an increase in impairment after controlling for demographic characteristics, perceived health, and the other disorders. A minority of patients with each disorder (range, 22.5% [substance use disorder] to 46.4% [panic disorder]) reported receiving mental health treatment in the last month. CONCLUSIONS: Clinically significant depression, anxiety, substance use, and suicidal ideation are quite common in this practice and associated with significant functional impairment. Primary care practices that serve poor urban immigrant populations have a critical need to provide access to mental health services. Arch Fam Med. 2000;9:876-883 PMID- 11031396 TI - SOAP: solutions to often asked problems. Chlamydia trachomatis testing. PMID- 11031397 TI - Barriers and benefits to leisure-time physical activity among older Mexican Americans. AB - OBJECTIVE: To identify the perceived barriers to and benefits of leisure-time physical activity among older Mexican Americans (MA) and European Americans (EA). DESIGN: Cross-sectional survey using in-home interviews of subjects. SETTING: Subjects recruited from 10 family practice offices in South Texas that are part of a practice-based research network. PARTICIPANTS: Two hundred ten MA and EA adults, aged 60 years and older, interviewed between April 1994 and April 1996. MEASUREMENTS: The perceived benefits and barriers summary score from the San Diego Health and Exercise Questionnaire, the Minnesota Leisure Time Physical Activity Questionnaire, body mass index, chronic diseases, depressive symptoms, and demographics. MAIN RESULTS: Older MA reported greater perceived benefits to physical activity and fewer perceived barriers than older MA while having lower levels of habitual physical activity. Lower levels of education, male sex, higher body mass index, and older age were also associated with lower levels of habitual physical activity. CONCLUSIONS: Although MA reported lower levels of physical activity, they perceived greater benefits and fewer barriers to physical activity. These attitudes about physical activity held by older MA may present an opportunity to encourage greater levels of physical activity throughout this population. PMID- 11031398 TI - Use of simvastatin treatment in patients with combined hyperlipidemia in clinical practice. For the Simvastatin Combined Hyperlipidemia Registry Group. AB - OBJECTIVE: To describe and understand current care of simvastatin-treated patients with combined hyperlipidemia in routine clinical practice. DESIGN: A 6 month prospective observational study. Demographics, simvastatin dosage, cardiac risk factors, and lipid profile were collected from August 1997 to December 1998 at 20 sites (230 patients) across the United States. RESULTS: Overall mean percentage of reduction in total cholesterol levels was 27% (P<.001), low-density lipoprotein cholesterol (LDL-C) was 35% (P<.001), and triglyceride values was 28% (P<.001). Among those patients with low baseline high-density lipoprotein cholesterol (HDL-C) values (<0.91 mmol/L [<35 mg/dL]) (N = 49), there was a 17% increase in HDL-C (P< or =.001); 35% of these patients achieved National Cholesterol Education Program HDL-C goal (ie, < or =0.91 mmol/L [> or =35 mg/dL]). Coronary heart disease (CHD) patients were given significantly higher initial doses (mean, 15.1 mg) compared with non-CHD patients (mean, 11.5 mg) (P< or =.001). Overall, 74% of patients achieved LDL-C goal (52% on starting dose, 22% after 1 titration). Among those patients who were not at goal and had a follow-up lipid profile result available, only 1 patient (2%) was at the maximum dose (80 mg); 69% were receiving 20 mg or less. Approximately 63% of patients with CHD, 80% of patients with 2 or more risk factors, and 91% of patients with fewer than 2 risk factors achieved LDL-C goal. CONCLUSIONS: Multiple factors contribute to LDL-C goal achievement in a usual care setting. A significant opportunity exists to increase the number of patients who achieve LDL-C goal by appropriate dose titration and/or give patients a higher initial dose of simvastatin. PMID- 11031399 TI - Tamoxifen's clinical applications: old and new. AB - The American Cancer Society estimates that this year more than 180, 000 women in the United States will develop breast cancer and more than 40,000 women will die of the disease. According to a National Cancer Institute model, 5 years of preventive therapy with tamoxifen citrate reduced the risk of invasive breast cancer by 49% (P<.00001) in women at increased risk for breast cancer. The reduction in risk was greater in women with a history of lobular carcinoma in situ (LCIS; 56% relative risk reduction) or atypical hyperplasia (86% relative risk reduction). It should be noted, however, that no benefit was found in 2 European studies using notably different risk evaluation models and entry criteria. Because elevated risks of uterine cancer and thromboembolic disease have been associated with tamoxifen therapy, appropriate counseling should be given to any woman considering tamoxifen as a means of reducing breast cancer risk. Arch Fam Med. 2000;9:906-912 PMID- 11031400 TI - Picornavirus infections: a primer for the practitioner. AB - Picornaviruses, including the rhinoviruses and enteroviruses, are common causes of infections in the developed world and the most common reason for prescribing antibiotics. These ubiquitous pathogens are increasingly being recognized in more serious illnesses, such as sinusitis, exacerbations of asthma, exacerbations of cystic fibrosis, myocarditis, meningitis, and severe neonatal sepsislike disease. Recent advances have improved our ability to diagnosis and treat these infections. PMID- 11031401 TI - A family physician's perspective on picornavirus infections in primary care. PMID- 11031402 TI - False-positive and indeterminate human immunodeficiency virus test results in pregnant women. AB - Guidelines from the Centers for Disease Control and Prevention, Atlanta, Ga, recommend that all pregnant women be offered human immunodeficiency virus (HIV) testing to ensure that they have the opportunity to use currently available therapeutic interventions to reduce the risk infecting their offspring with HIV. These recommendations have resulted in an increased number of low-risk women being tested and a significant rise in the percentage of false-positive results from HIV antibody screening tests and ambiguous (indeterminate) findings from confirmatory tests. Women receiving such results are generally in emotional turmoil yet must make treatment choices if they prove to be infected. This article provides guidelines to help general medical practitioners to understand the nature of HIV testing, to assess a woman's infection status when initial tests are ambiguous, and to determine when treatment is appropriate. PMID- 11031404 TI - Improving quality or shifting diagnoses? What happens when antibiotic prescribing is reduced for acute bronchitis? AB - BACKGROUND: A quality improvement project in an academic practice demonstrated a reduction in antibiotic prescribing for acute bronchitis. However, it was unclear whether this represented a reduction in antibiotic use or whether physicians assigned new diagnoses to the same patients to avoid scrutiny and continue to use antibiotic therapy. OBJECTIVE: To examine whether a substantial amount of diagnostic shifting occurred while antibiotic prescribing for acute bronchitis decreased during a 14-month period (from January 1, 1996, to February 28, 1997). METHODS: All patient diagnoses of acute bronchitis, acute sinusitis, upper respiratory tract infection, and pneumonia were determined for the 14 months of the acute bronchitis intervention. The relative distribution of patients among these 4 diagnostic categories was compared to determine if the percentage of patients with acute bronchitis decreased while those with acute sinusitis and pneumonia increased during the acute bronchitis intervention. RESULTS: The percentage of patients with the diagnosis of acute bronchitis remained unchanged during the 14-month period while antibiotic use for this condition decreased from 66% of cases to less than 21% of cases. Instead of the patients being assigned a different diagnosis such as acute sinusitis so that antibiotic prescribing would not be scrutinized, as we hypothesized, the relative number of diagnoses for acute sinusitis compared with acute bronchitis actually declined during the 14 months. No change was noted in the relative frequency of acute bronchitis cases compared with pneumonia cases. CONCLUSION: During a 14-month period when an intervention was successful at reducing antibiotic use for acute bronchitis, there was no evidence that physicians shifted patients from the diagnosis of acute bronchitis to other diagnoses. PMID- 11031403 TI - Are antibiotics necessary in the treatment of locally infected ingrown toenails? AB - CONTEXT: A wide variety of generalists and specialists treat locally infected ingrown toenails, with perhaps the most common treatment regimen including resection of the nail border coupled with oral antibiotics. OBJECTIVE: To determine whether oral antibiotic therapy is beneficial as an adjunct to the phenol chemical matrixectomy in the treatment of infected ingrown toenails. DESIGN: We prospectively enrolled healthy patients with infected ingrown toenails. Each patient was randomly assigned to 1 of 3 groups that received either 1 week of antibiotics and a chemical matrixectomy simultaneously (group 1), antibiotics for 1 week and then a matrixectomy (group 2), or a matrixectomy alone (group 3). SETTING: Institutional ambulatory outpatient clinic. PATIENTS: Fifty-four healthy patients with infected ingrown toenails were studied. Patients with immunocompromised states, peripheral vascular disease, or cellulitis proximal to the hallux interphalangeal joint were excluded. Groups were age matched for comparison. RESULTS: Mean healing times for groups 1, 2, and 3 were 1.9, 2.3, and 2.0 weeks, respectively. Subjects receiving antibiotics and a simultaneous chemical matrixectomy (group 1) healed significantly sooner than those receiving a 1-week course of antibiotics followed by a matrixectomy (group 2). There was not a significant difference in healing time between those that received a chemical matrixectomy alone (group 3) and those that received a matrixectomy coupled with a course of oral antibiotics (group 1). CONCLUSION: The use of oral antibiotics as an adjunctive therapy in treating ingrown toenails does not play a role in decreasing the healing time or postprocedure morbidity. PMID- 11031405 TI - Comparison of the laser and phenol chemical peel in facial skin resurfacing. AB - OBJECTIVE: To determine differences in postoperative outcomes, complications, and adverse effects between phenol chemical peel (CP) and the carbon dioxide laser peel, when used for facial skin resurfacing. DESIGN: Nonrandomized prospective comparison of 2 facial skin resurfacing techniques using a split-face paradigm. In this initial study, 18 months of follow-up data are available, including the patients' subjective evaluations, the surgeons' objective assessments, and a histological analysis of 1 patient by a blinded pathologist. SETTING: A facial plastic surgery clinic associated with a university medical center. PATIENTS: Four female patients with actinic-damaged facial skin and facial rhytids, aged 61 to 73 years. INTERVENTIONS: The left side of each face was treated with a phenol based CP formula according to standard procedure. The right side was resurfaced using the Sharplan Silktouch Flashscanner carbon dioxide laser. Patients were photographed before treatment and at regular intervals postoperatively. One patient underwent rhytidectomy at 2 months posttreatment, and specimens were obtained for histological analysis. MAIN OUTCOME MEASURES: Evaluation of observable clinical improvement in skin quality, postoperative swelling, erythema, pigmentary alterations, healing time, and complications. RESULTS: All 4 patients experienced transient initial discomfort on the CP side that subsided within 24 hours after treatment. The laser side was noted to have slightly more prolonged stinging, erythema, and edema. Erythema was noted to be more uniform in the laser-treated areas. Final clinical improvement in rhytids was evaluated by 4 surgeons who reviewed color slide presentations of each patient 1 year or more postoperatively. Uniform wrinkle improvement was noted around the eyelid and lateral cheek areas on both the CP and laser-treated sides. A moderate advantage in the degree of wrinkle improvement was noted on the laser-treated sides of the upper lip and forehead. Thick-skinned, glandular skin areas, such as the nasolabial fold and chin, were found to be substantially smoother in the laser treated areas. Histological studies indicate that the CP side was noted to have a deeper injury, extending into the reticular dermis. The skin treated with the laser was injured more superficially, down to the papillary dermis. CONCLUSIONS: Phenol CP is as effective as the laser in diminishing rhytids in the thin-skinned areas of the face. The laser produces improved results in the thick, glandular areas of the face, but also produces more intense hypopigmentation, longer periods of patient discomfort, and longer periods of postoperative erythema. Both phenol CP and laser resurfacing remain useful clinical tools. PMID- 11031406 TI - Upper respiratory morbidity in preschool children: a cross-sectional study. AB - OBJECTIVES: To estimate the occurrence of and associations between upper respiratory tract infections in preschool children, and to assess constitutional and environmental factors as determinants of these infections. DESIGN: Population based cross-sectional study. SETTING: Oslo, Norway. PARTICIPANTS: Preschool children, aged 4 to 5 years (3853 completed questionnaires). MAIN OUTCOME MEASURES: Acute and recurrent acute otitis media, tonsillopharyngitis, common cold, and rhinitis. RESULTS: Upper respiratory tract infections were common at age 4 years. During the last month, 7.1% of the children had acute otitis media and 7.5% experienced tonsillopharyngitis. Corresponding figures for common cold and rhinitis were 58.3% and 16. 4%, respectively. During the last 12 months, 9.5% of the children experienced more than 1 bout of acute otitis media, 6.9% had more than 1 tonsillopharyngitis episode, 47.7% contracted more than 2 common colds, and 3.2% had rhinitis weekly or monthly. The lifetime prevalence of recurrent acute otitis media (>/=4 episodes in any 12-month period) was 12.7% (n = 473). Bivariate correlations showed small-to-moderate relationships between the infections. Common cold was only weakly related to otitis media, tonsillopharyngitis, and chronic rhinitis. The probability for developing acute otitis media was almost 4-fold increased in children who had tonsillopharyngitis in the last year (adjusted odds ratio = 4.19; 95% confidence interval, 3.09 5.66). In logistic regression analysis, atopic disease was a strong determinant of all upper respiratory tract infections. Low birth weight increased the risk of acute otitis media. Day care attendance and the presence of siblings, which were considered to be indicators of exposure to respiratory pathogens, increased the risk of upper respiratory tract infections. CONCLUSIONS: In preschool children, acute otitis media, tonsillopharyngitis, and common cold were quite common, while chronic rhinitis was less prevalent and strongly associated with atopic disease. Attendance at day care centers increased the risk of upper respiratory tract infections in this age group, although the effect was weaker than that in younger children. PMID- 11031407 TI - Results of larynx preservation surgery for advanced laryngeal cancer through tracheal autotransplantation. AB - OBJECTIVE: To evaluate the use of tracheal autotransplantation for reconstruction of the hemilaryngectomy defect that includes the hemicricoid cartilage and results from resection of laryngeal or hypopharyngeal cancer. DESIGN: The clinical records of 28 patients undergoing primary or salvage hemicrico hemilaryngectomy for laryngeal (26 patients) and pyriform sinus (2 patients) cancer were analyzed for function and local control. SETTING: Academic center. PATIENTS: Case series review of 28 consecutive patients treated during a 3(1/2) year period who had an average follow-up period of 19 months. INTERVENTION: Twenty-five men and 3 women, aged from 28 to 79 years, underwent a hemilaryngectomy that included the hemicricoid cartilage, the ipsilateral thyroid lobe, and a unilateral or bilateral lymph node dissection. In 26 patients, these extensive defects were reconstructed with a tracheal autotransplantation that restored the larynx at the glottic and subglottic levels. In 2 patients, the defect was converted into a total laryngectomy because of tumor extension beyond the resection margins of a hemicrico-hemilaryngectomy. MAIN OUTCOME MEASURES: The times to decannulation and retake of full oral feeding, the quality of speech, and the incidence and site of recurrent cancer were assessed. RESULTS: Of the 26 patients undergoing tracheal autotransplantation, 24 were decannulated and all regained the ability to maintain nutrition by mouth. Ultimate voice quality was "subnormal" to "moderately hoarse" in all patients. Five recurrences developed of which 2 were treated with total laryngectomy. CONCLUSIONS: Functional reconstruction of extensive laryngeal defects can be achieved with an autotransplantation of cervical trachea, with favorable functional results and acceptable morbidity. This technique expands the limits of conservation surgery for selected laryngeal and hypopharyngeal tumors. PMID- 11031408 TI - Role of CD44 variant exon 6 in invasion of head and neck squamous cell carcinoma. AB - OBJECTIVES: To determine the correlation between the expression of CD44 variant exon 6 (v6) and the clinicopathological features of head and neck squamous cell carcinomas (HNSCCs), and to study the role of CD44v6 in cell invasion using a human HNSCC cell line (HSC-2). DESIGN: The expression of CD44v6 was evaluated using immunohistochemical analysis in paraffin-embedded tissue specimens from 89 primary lesions. The concentration of CD44v6 protein in 37 cryopreserved tumor specimens was evaluated using the enzyme-linked immunosorbent assay. The HSC-2 cells were treated with 2F10, a monoclonal antibody against CD44v6. The effects of 2F10 on HSC-2 cell proliferation, migration, and invasion potential were evaluated. RESULTS: The down-regulation of CD44v6 expression or the concentration of cancer tissue significantly correlated with a lower degree of pathohistological differentiation and a higher rate of cervical metastasis. The invasion of HSC-2 cells into type I collagen gel and the expression of CD44v6 were decreased in invading cells released from the upper layer. Furthermore, the treatment of HSC-2 cells with 2F10 significantly enhanced cell invasion. However, 2F10 did not affect either the proliferation or migration properties of HSC-2 cells. CONCLUSIONS: The down-regulation of CD44v6 expression may be useful as a biological marker for the degree of malignancy in HNSCCs. We assume that the loss or dysfunction of CD44v6 is involved in the acquisition of invasion ability in HSC-2 cells. In addition, the potential existence of a CD44v6-mediated signal transduction pathway may play a role in inhibiting the invasion in HNSCCs. PMID- 11031410 TI - Influence of growth factors on tissue-engineered pediatric elastic cartilage. AB - OBJECTIVE: To investigate the influence of growth factors on tissue-engineered pediatric human elastic cartilage relative to potential clinical application. DESIGN: Controlled study. SUBJECTS: Eleven children ranging in age from 5 to 15 years provided auricular elastic cartilage specimens measuring approximately 1 x 1 x 0.2 cm and weighing approximately 100 mg. INTERVENTIONS: Three million chondrocytes were plated into 4 groups of Ham F-12 culture medium: group 1, Ham F 12 culture medium only; no growth factors (control group); group 2, Ham F-12 culture medium and basic fibroblast growth factor; group 3, Ham F-12 culture medium and transforming growth actor beta; and group 4, Ham F-12 culture medium and a combination of both growth factors. At 3 weeks, the cells were harvested and mixed with a copolymer gel of polyethylene glycol and polypropylene oxide (Pluronic F-127). The cell solution was injected subcutaneously into athymic mice. The constructs were harvested at up to 22 weeks of in vivo culture and histologically analyzed. RESULTS: The average number of cells generated in vitro was as follows: group 1, 12 million; group 2, 40 million; group 3, 7 million; and group 4, 35 million. Group 2 in vivo gross specimens were the largest and heaviest. Histologically, the control group and the basic fibroblast growth factor group (groups 1 and 2) exhibited characteristics compatible with normal auricular cartilage; groups 3 and 4 demonstrated cellular disorganization and moderate to severe fibrous tissue infiltration. CONCLUSIONS: Basic fibroblast growth factor demonstrates the greatest positive influence on the in vitro and in vivo growth of engineered pediatric human auricular cartilage. The results suggest that basic fibroblast growth factor has the potential for clinical application in which a goal will be to generate a large volume of tissue engineered cartilage from a small donor specimen in a short period of time and of a quality similar to native human elastic cartilage. PMID- 11031409 TI - Systemic adoptive T-cell immunotherapy in recurrent and metastatic carcinoma of the head and neck: a phase 1 study. AB - OBJECTIVE: To evaluate the feasibility and toxic effects of systemic adoptive T cell immunotherapy in patients with unresectable squamous cell carcinoma of the head and neck (SCCHN). DESIGN: Nonrandomized phase 1 clinical trial. SETTING: Academic tertiary care hospital. PATIENTS: Between April 1, 1996, and September 30, 1998, 17 patients with confirmed recurrent and metastatic SCC of the upper aerodigestive tract were enrolled. Two patients did not receive T cells because of poor vaccine response. Fifteen patients were successfully treated with T-cell immunotherapy. INTERVENTION: Patients were vaccinated on the thigh with irradiated autologous tumor cells admixed with granulocyte-macrophage colony stimulating factor (GM-CSF) followed by 3 additional daily injections of GM-CSF at the vaccination site. Eight to 10 days later, tumor cell vaccine-draining inguinal lymph nodes were resected, and lymph node lymphocytes were activated with staphylococcal enterotoxin A and expanded in interleukin 2 in vitro. Resulting cultured cells were infused into patients peripherally on an outpatient basis. RESULTS: Toxic effects of infusion were limited to grade 2 reactions in 3 of 16 treatments. One patient required overnight hospitalization for fever and emesis. Median cell expansion was 37 times (range, 4-416 times), and median cell dose was 7.5 x 10(9) (range, 1.3 x 10(8) to 4.2 x 10(10)). Infused cells were predominantly CD3+ (>97%), being a mixture of CD4+ and CD8+ cells. Three patients demonstrated stabilization of previously progressive disease. Two patients experienced favorable clinical courses after adoptive T-cell transfer, including 1 patient with no evidence of disease 4 years after surgical resection of a vertebral body metastasis. CONCLUSIONS: Adoptive immunotherapy is a technically feasible and safe treatment with low toxicity and may demonstrate therapeutic activity in patients with unresectable SCCHN. PMID- 11031411 TI - Clinical update on 10 children treated with intralesional cidofovir injections for severe recurrent respiratory papillomatosis. AB - OBJECTIVES: To continue assessment of the benefits and risks of intralesional administration of cidofovir, an acyclic nucleoside phosphonate, for treating severe recurrent respiratory papillomatosis (RRP) in pediatric patients, and to discuss guidelines for larger prospective multi-institutional studies of the use of cidofovir. DESIGN: Prospective case series. SETTING: Tertiary care children's hospital. PATIENTS: A total of 10 patients with severe RRP (defined as requiring debulking procedures to maintain airway patency at least once a month) underwent intralesional cidofovir therapy. The original 5 patients have received more than 1 year of follow-up since their last cidofovir injection, and 5 subsequent patients have been treated with a revised injection protocol. INTERVENTION: Microsuspension laryngoscopy with intralesional injection of cidofovir after repetitive carbon dioxide laser treatments and mechanical debulking of papillomas. MAIN OUTCOME MEASURES: Papilloma stage at the time of serial laryngoscopies. Histologic examination of biopsy specimens of laryngeal tissue obtained 1 year or more after last cidofovir injection. RESULTS: There was evidence of marked improvement in the 4 of the 5 new patients enrolled under the revised injection protocol, continuation of a disease-free state in 1 of the original 5 patients, and sustained improvement in 4 of the 5 original patients, resulting in a significantly reduced interval of intervention. CONCLUSIONS: Intralesional cidofovir therapy continues to show benefit in the treatment of severe RRP in pediatric patients. Safety profiles have not been fully established, but current histologic data are reassuring. PMID- 11031412 TI - Notch signaling and the emergence of auditory hair cells. AB - OBJECTIVE: Recent insights into the mechanisms that determine a hair cell's fate have emerged from studies on invertebrate sensory organs and the avian inner ear. These mechanisms have important implications for our understanding of the possible therapeutic management of sensorineural deafness. This article reviews the current state of our knowledge regarding mammalian auditory hair cell fate specification. DESIGN: Data were obtained from the MEDLINE database and data presented at the Molecular Biology of Hearing and Deafness Meeting (Bethesda, Md, October 1998). Articles reporting information about cell fate specification and Notch and its ligands were selected. MAIN OUTCOME MEASURES: Data pertaining to cell fate mechanisms, Notch and its ligands, and application to hearing were extracted. RESULTS: The Notch/ligand mechanism is responsible for the specification of the hair cell phenotype. CONCLUSIONS: Major progress has been made in understanding this fundamental process, and its application to hair cell determination is only now being realized. Possible applications could involve the "switching" of supporting cells to hair cells, thus replenishing those hair cells damaged in sensorineural hearing loss. PMID- 11031413 TI - Audiometric configurations following exposure to explosions. AB - OBJECTIVE: To determine the configurations in pathologic audiograms obtained in patients shortly after exposure to an explosion. DESIGN, SETTING, AND PATIENTS: Audiograms were performed in 143 patients (286 ears; 76 males and 67 females, with a mean age of 34.6 years [range, 11-79 years]) sent to the Department of Otolaryngology and Head and Neck Surgery in a city hospital located in the center of Jerusalem, Israel, after being injured in 4 severe explosions occurring in Jerusalem during 1995-1997. Most of the audiograms were obtained within 4 hours after the explosion, and the remaining were obtained within 4 days of the explosion. RESULTS: Of 200 pathologic audiograms, 93 (46%) showed a downward slope configuration, 82 (41%) showed a dip configuration, and 25 (12%) were flat. There were 38 audiograms (19%) with 6-kHz dips. In 82% of the patients, the audiometric configurations were similar in both ears. Patients with a slope configuration on the audiogram were significantly older than those with a dip configuration (mean age, 40.8 vs 32.8 years; P<.01). CONCLUSIONS: There does not appear to be a single typical audiometric configuration in patients exposed to explosions. The slope and dip configurations are most frequently seen and are approximately equal in their incidence. This observation may lead to better understanding damage to the inner ear as a result of an explosion. This study is distinctive because of the large number of audiograms obtained and the fact that most of them were obtained immediately after the explosion. PMID- 11031414 TI - Endovascular treatment of acute and subacute hemorrhage in the head and neck. AB - OBJECTIVE: Acute and subacute hemorrhage in the head and neck often represent a life-threatening situation. The goal of this study is to evaluate the indications for and contributions of endovascular techniques in the diagnosis and management of such severe cases. DESIGN: Seventy-two patients with acute or subacute intractable hemorrhage of the head and neck were treated over a period of 5 years: 2 patients had experienced trauma; in 6 cases the cause of bleeding was iatrogenic; and in 2 patients intraosseous arteriovenous malformations were manifested. Fifteen patients had tumors, 9 of whom had prior radiotherapy. Forty seven patients presented with epistaxis (41 idiopathic and 6 during anticoagulation therapy). The endovascular therapy was performed using polyvinyl alcohol particles, fibered platinum or electrolytically detachable coils (Guglielmi detachable coils; Target Therapeutics, Fremont, Calif), a stent, glue (Ethibloc; Ethicon GmbH, Norderstedt, Germany, and Histoacryl; B. Braun Melsungen AG, Melsungen, Germany), or with a combination of these different embolic materials. RESULTS: The acute bleeding was successfully controlled in all cases. Fourteen patients (7 with epistaxis, 5 with tumors, and 2 with arteriovenous malformations) had to be embolized more than once before the bleeding could be controlled. The idiopathic, traumatic, iatrogenic, and remaining tumoral cases were treated only once. The long-term morbidity was 1.9%. CONCLUSIONS: Owing to the recent continuous advances in interventional radiologic techniques, it is possible to treat both acute and subacute life-threatening head and neck hemorrhage most efficiently. In many cases the endovascular therapy complements surgery. PMID- 11031415 TI - Imaging quiz case 1. Grade 1 chondrosarcoma (CS) of the larynx. PMID- 11031416 TI - Imaging quiz case 2. Cavernous hemangioma. PMID- 11031417 TI - Imaging quiz case 3. Laryngeal schwannoma. PMID- 11031418 TI - Imaging quiz case 4. Dentigerous cyst. PMID- 11031419 TI - Directed functional endoscopic sinus surgery and headaches. PMID- 11031420 TI - Directed functional endoscopic sinus surgery and headaches. PMID- 11031422 TI - The 4th annual scientific meeting of the Japanese Heart Failure Society. October 8-10, 2000, Kobe, Japan. Abstracts. PMID- 11031421 TI - Directed functional endoscopic sinus surgery and headaches. PMID- 11031423 TI - [Discrimination between the effects of hypolipidemic and antiatherogenic drugs: probucol pharmacokinetics (review of the literature)]. PMID- 11031424 TI - [Diagnosis of short-term subclinical shifts in the level of humoral parameters]. AB - Algorithm of evaluating the blood humoral parameters has been theoretically and experimentally validated and clinically tried with due consideration for the circaseptan fluctuations in their levels. Probability of deviations from the normal range of values depends on the number of measurements making use of the suggested algorithm. PMID- 11031425 TI - [State of the art and prospects of laboratory diagnosis of streptococcal infections in Russia]. PMID- 11031426 TI - [Laboratory diagnosis of cytomegalovirus infection]. PMID- 11031427 TI - [Detection of Staphylococcus aureus and Staphylococcus epidermis species specific antigens and antibodies to alpha-toxin in the blood of patients with pneumonia]. AB - Test systems for indirect hemagglutination (IHA) test for detection of S. aureus and S. epidermidis teichoic acids and S. aureus alpha-toxin in patients' sera have been developed on the basis of immunoglobulins isolated from monospecific sera. Test system for IHA test for detection of antitoxin in donor and patients' sera has been created on the basis of highly purified alpha-toxin. Thirty donor sera and 61 sera from patients with pneumonia were analyzed. Low antibody levels in the patients may be due to the fact that the sera were collected during the first days of disease. Group of patients with high content of staphylococcal antigens and antitoxin in the blood was particularly interesting. These patients developed severe pneumonia, among whose etiological agents were S. aureus and S. epidermidis. Diagnostic analysis of patients' sera by IHA test for detection of staphylococcal antigens was more effective, accurate, and rapid in comparison with the bacteriological method; moreover, it confirmed the significance of staphylococci in the pathogenesis of pneumonia. PMID- 11031428 TI - [Molecular genetic diagnosis of streptococcal, pertussis and parapertussis infections]. PMID- 11031429 TI - [Algorithm for specific laboratory diagnosis in viral hepatitis among inpatients (lecture)]. PMID- 11031430 TI - [Development of an original computer program FISHMet: use for molecular cytogenetic diagnosis and genome mapping by fluorescent in situ hybridization (FISH)]. AB - Original software FISHMet has been developed and tried for improving the efficiency of diagnosis of hereditary diseases caused by chromosome aberrations and for chromosome mapping by fluorescent in situ hybridization (FISH) method. The program allows creation and analysis of pseudocolor chromosome images and hybridization signals in the Windows 95 system, allows computer analysis and editing of the results of pseudocolor hybridization in situ, including successive imposition of initial black-and-white images created using fluorescent filters (blue, green, and red), and editing of each image individually or of a summary pseudocolor image in BMP, TIFF, and JPEG formats. Components of image computer analysis system (LOMO, Leitz Ortoplan, and Axioplan fluorescent microscopes, COHU 4910 and Sanyo VCB-3512P CCD cameras, Miro-Video, Scion LG-3 and VG-5 image capture maps, and Pentium 100 and Pentium 200 computers) and specialized software for image capture and visualization (Scion Image PC and Video-Cup) have been used with good results in the study. PMID- 11031431 TI - [Current methods of molecular cytogenetics in pre- and postnatal diagnosis of chromosome aberrations]. AB - Progress in prevention of chromosome aberrations is due to utilization of molecular cytogenetic diagnostic methods. The purpose of this trend of clinical cytogenetics is development and utilization of new highly effective methods for analysis of chromosome aberrations. Molecular cytogenetic methods (fluorescent in situ hybridization-FISH) are used for pre- and postnatal identification of chromosome aberrations in mentally retarded children and congenital diseases. These studies are carried out after classical cytogenetic analysis, if it proves to be of no avail. FISH diagnosis pre- and postnatally detects autosomal trisomy, gonosome aneuploidy (including mosaic forms), marker chromosomes, structural chromosome aberrations, including fragile X chromosome syndrome. Rapid (15-30 min) FISH with an original collection of centromere, telomere, and site-specific DNA probes (plasmid, cosmid, PAC and YAC clones) is recommended for molecular cytogenetic diagnosis. FISH diagnosis is an effective complex of methods for pre- and postnatal identification of chromosome aberrations and a necessary supplement to classical cytogenetic diagnosis. Molecular studies of chromosome aberrations are significant for theoretical and applied studies, for they help detect patients with specific chromosome syndromes from a vast group of children with undifferentiated mental retardation and congenital diseases. PMID- 11031432 TI - [Laboratory tests in the assessment of prognosis in HIV infection]. PMID- 11031433 TI - [Diagnosis of opportunistic Candida infection]. PMID- 11031434 TI - [Inhibition of DNA amplification at individual genome sites in avirulent Mycoplasma pneumoniae strains]. PMID- 11031435 TI - [Detection of toxin-producing pathogenic bacterial strains by polymerase chain reaction]. AB - Polymerase chain reaction (PCR) was used for detection of pathogenic Clostridium botulinum, Clostridium perfringens, Clostridium difficile, and Escherichia coli. With this aim in view, primers to botulinic toxins types A, B, C1, D, E, F, and G, perfringens enterotoxin, difficile toxin, and types 1 and 2 Shigella-like toxins were chosen and synthesized. Optimal amplification conditions were selected for each pair of primers, with DNA and the respective agent as the reaction mixture matrices. PCR was highly specific and sensitive in all cases. Its sensitivity was 10-100 cells/sample. Among the tested C. botulinum, C. perfringens, C. difficile, and E. coli strains, specific amplification products of expected size were observed only in the strains containing the respective toxin genes. These findings recommend the use of these methods in clinical microbiology. Strains containing type 2 Shigella-like toxin gene were detected among E. coli strains isolated from patients with the hemolytic uremic syndrome, which for the first time indicates that the problem with E. coli epidemic strain O157 is valid for Russia. As a result of our studies, test systems for detection of types A, B, C, D, E, F, and G C. botulinum strains, C. perfringens and C. difficile, and E. coli O157 strains are now available. PMID- 11031436 TI - [Laboratory diagnosis of mycoplasma infections in man]. PMID- 11031437 TI - [The status of and developmental outlook for sports medicine and therapeutic physical exercise in Ukraine]. AB - The sports medicine service in Ukraine, its structure, priority tasks, manpower potential, extent and quality of care-giving to sportsmen and those individuals engaged in physical culture over the period 1997-1998 are detailed. Case follow up findings are submitted together with indices for medical and biological supplies to sportsmen. Ways for further improvement of the sports medicine service are outlined. PMID- 11031438 TI - [The immunopathogenesis of psoriatic disease]. PMID- 11031439 TI - [The cytogenetic assessment of the degree of severity of acute radiation sickness in those who cleaned up the Chernobyl catastrophe]. AB - The study is of a retrospective character and is based on the author's own cytogenetic findings secured in May-June 1986 in the examination of patients diagnosed as having had a first to third-degree acute radiation sickness while receiving massive detoxicating therapy. We consider it expedient to employ the model of multiple linear regression relying on a complex of most informative cytogenetic indicators rather than on only one such indicator alongside the traditional dose reaction identifiable by the level of dicentric chromosomes in blood lymphocytes. This will allow the degree of severity of the radiation injury to be determined with greater precision and permits assessing the therapeutic efficiency of the measures implemented. PMID- 11031440 TI - [The treatment experience of rheumatoid arthritis patients in an industrial region of the Donets Basin]. AB - The data submitted suggest that the most effective option in the treatment of rheumatoid arthritis is simultaneous employment of a nonsteroidal anti inflammatory agent and metatrexate for 1.5 yr and courses of hormonotherapy, exercise therapy, and topical application to the joints of compresses with dimexide, heparin, and analginum as required and ultrasound with hydrocortisone as well. There was no increase in disability rates among patients with rheumatoid arthritis placed on the above combined treatment, which fact is also related to improvement of diagnosis rheumatoid arthritis, well-timed case follow-up, and prescription of courses of prophylactic treatment. PMID- 11031441 TI - [The anisotropy of the skin of the precordial area in patients with stable and unstable stenocardia]. AB - Coronary patients were studied for the precordial area skin anisotropic properties with the aid of EHF differential-diagnostic reflectometry. Disclosed in the patients were the following points of anisotropia: E18S in the 5th intercostal space along the medioclavicular line on the left, E18D--a symmetric point on the right, R24S in the 3d intercostal space along the parasternal line on the left, R24D--a symmetric point on the right. The value for anisotropia determined by anisotropia factor in decibels will rest with the degree of abnormal changes on the cardiac muscle, which fact has been documented by electrocardiographic findings. PMID- 11031442 TI - [The psychological personality characteristics of young hypertension patients]. AB - Overall forty-eight patients with stage I-II hypertensive disease (HD) were examined. A psychological investigation in the patients was conducted as recommended by Eysenck [correction of Isenk], S. Rosenzweig, Luchier. HD patients presenting with aggravated heredity (AH) and their HD parents are more often than not inclined to introversion. In AH-free HD patients, a high level of neuroticism was recordable as was decrement in their ability to compensate anxiety and a low level of social adaptation. It is suggested that AH HD patients might have some mechanism of compensation of anxiety early in the course of the illness. PMID- 11031443 TI - [Meteoropathic reactions in patients with circulatory system diseases: their prevalence, the characteristics of their manifestation and their correction]. AB - Presented in the paper are topical aspects of prevalence, specificities of manifestation, and correction of meteoropathologic reactions (MR) in disorders of the circulatory system. A total of 1260 patients were examined. Of these, 518 had hypertensive disease (HD), 198 were coronary (IHD) patients, 544 had HD concurrent with IHD. Based on the standardized questionnaire and results of clinical and instrumental studies meteorosensitivity was assessed, with meteorogramme having been taken into account. To correct MR aero- and balneoprocedures were used together with speleotherapy and physical training. High meteorologic factors sensitivity was ascertained in 67 percent of patients. MR frequency was found out to be related to overstrain, failure to stick to the daily time-table, bad habits, constitutional specificities, low physical activity. Depending on the features of manifestation three degrees of severity have come to be recognized in MR. A complex of measures designed to correct MR is submitted. The use of the above complex allowed the MR frequency to be reduced, their severity to be alleviated. PMID- 11031444 TI - [Intrathoracic compressions of the respiratory tract due to a double aortic arch in children]. AB - Overall, twenty-eight children with intrathoracic airway compressions (IAC) caused by double aortic arch (DAA), who ranged 1.5 months to eleven years old, were kept under medical surveillance. The following respiratory symptoms were found to dominate the clinical picture in the above pediatric patient population: stridor, dyspnea, bouts of asphyxia, orthopnea, with secondary bronchopulmonary complications having been disclosed in all. The diagnosis of IAC and its complications in DAA was based on findings from thoracic roentgenography, esophagography, digital subtraction angiography, and tracheobronchoscopy. This pathology surgical correction indications were unquestionable. The policy has been worked out of treating DAA-associated IAK through surgery, with particular regard to the anatomical variants of the defect under question and its complications. PMID- 11031445 TI - [The creation of a "new" aorta and the characteristics of coronary artery transplantation in the anatomical correction of transposition of the great vessels of the heart]. AB - Opinion varies among surgeons regarding the preferable method of surgical correction in patients with transposition of the great vessels (TGV) presenting with abnormal origins of the coronary artery (CA) and inadequacies in the diameters of the great vessels. In the work presented, the experience is analyzed gained with the performing of TGV radical correction operations of the arterial switch type in patients presenting with anomalous CA origins and abnormalities in diameters of the great vessels (n = 5). The submitted option permits simplifying CA transfer avoiding the coronary blood flow-related fatal intraoperative complications. Our modification of creation of a "new" aorta combined with particular features of implantation of the coronary arteries enabled us to secure good results in the presence of malformed, TGV, anatomy, which fact allowed the arterial switch operations to be extended to a greater number of cases in disadvantageous anatomical variants of TGV. PMID- 11031446 TI - [The morphological characteristics of apoptosis and mitotic activity of the gastric epitheliocytes in patients with nonatrophic and atrophic forms of chronic gastritis]. AB - Overall, 142 patients of both sexes with non-atrophic and atrophic forms of chronic gastritis who ranged from 17 to 65 years old and 25 healthy subjects (control group) were studied for apoptotic activity and mitotic patterns of gastric epitheliocytes (GE). Progression of chronic gastritis was found out to be associated with intensification of GE mitotic activity and apoptosis. The studied parameters abnormal changes were at their greatest in patients with atrophic and hyperplastic forms of chronic gastritis. PMID- 11031447 TI - [The effect of certain factors on the prognosis of the course of acute pancreatitis]. AB - An analysis of treatment of 2740 patients with acute destructive pancreatitis from 1973 up to the present has been performed together with that of relevant published literature. Different factors of diagnosis and those of treatment of acute pancreatitis were found to influence the prognosis of the illness in some way or other. It has been noted that delayed surgery permit obtaining better results. Data are presented on long-term results of medical and surgical treatment of those patients who had become ill with necrotic forms of acute pancreatitis. PMID- 11031448 TI - [The psychosomatic correlations and the antibodies to lung tissue in patients with bronchial asthma]. AB - Forty-four patients with bronchial asthma, aside from clinical and functional evaluation, were studied for the psychological profile (MMPI-test) and the presence of antibodies to the lung tissue. The correlations established were as follows: normal titers of antibodies to the lung tissue--more agreeable and satisfactory psychological profile of the patients, the lower or higher the antibody titer, the less acceptable is the psychological profile. PMID- 11031449 TI - [The local immunity status of patients with chronic dust-induced bronchitis]. AB - The ratio of cells in the bronchoalveolar washing was studied as was the functional state of phagocytizing cells, with assessment thereof having been done in the pathogenesis of chronic bronchitis in those workers exposed to quartz containing dust. The conducted studies showed the endopulmonary cytogram to be characterized by a high degree alveolar neutrophilia (28.2 +/- 1.3) in chronic bronchitis. Disturbances in local immunity were also evidenced by the dysfunction of alveolar macrophages (decline in adhesiveness and absorptivity against the background of apparent activation of intracellular oxygen-dependent metabolism), and by the decrease of the level of secretory immunoglobuline A. PMID- 11031450 TI - [The degree of the severity of the course of acute lung diseases in newborn infants in the early neonatal period]. AB - In a prospective clinical investigation, a comparative evaluation was done of degrees of severity of most commonly encountered acute lung disorders in 95 newborn infants who had been treated in the Lviv Regional Neonatal Centre. It has been ascertained that particular features of the course of acute lung diseases in early neonatal period are determined by gravity of the affection of the lung that can be assessed with the aid of the calculable index. The newborn babies aged 11 to 23 h presenting with respiratory distress syndrome displayed significant differences in values for the lung affection severity index, the same being true of those children having been dying at a later age or surviving with subsequent bronchopulmonary dysplasia being formed. The above index can be used in the differential diagnosis of lung diseases, it has also prognostic significance for assessment of risk of death in a child or that of subsequent development of bronchopulmonary dysplasia. PMID- 11031451 TI - [The coagulation link of the hemostatic system in newborn infants from mothers with extragenital pathology]. AB - Overall, fifty newborn infants born to mothers with extragenital pathology (diabetes mellitus) were studied for the coagulative link of the homeostasis system as were 20 babies born to essentially healthy mothers. The studies were made by day 1 to 3 and 5 to 7 the babies' life. The conducted studies revealed signs of hypercoagulation presenting with activation of the external and internal route of hemostasis in those infants born to mothers with diabetes mellitus (DM). Toward the period of early neonatal adaptation shifts are still observable in the coagulating system at the expense of stimulation of the first phase of coagulation. The disclosed abnormalities in the coagulative link of the coagulating system in infants born to DM mothers pose a threat of development of thrombohaemorrhagic complications. PMID- 11031452 TI - [The toxicosis parameters in children with different diseases]. AB - A technique is submitted of separation of blood serum into constituent fractions with subsequent assaying of cytolytic activity (toxicity) of each of the blood substances relative to autologous leukocytes. The elaborated techniques for the study into toxicity permits not only ascertaining the presence of toxicosis together with degree of its severity but also gives the clinician an opportunity to individualize the multicomponent detoxicating therapy indications. PMID- 11031453 TI - [The ultrastructural characteristics of the early diagnosis of mycosis fungoides]. AB - As a result of studies made (biopsy specimens of damaged skin from seven patients presenting with the erythrodermic stage of mycosis fungoides) a conclusion was reached that at the given stage the following two processes take place: (1) a rise in the proliferative activity of keratinocytes and (2) disturbances their differentiation. These ultrastructural manifestations can serve as criteria for a well-timed diagnosis of mycosis fungoides to be made. PMID- 11031454 TI - [The morphometric and strength characteristics of the femur for the biomechanical validation of blocking intramedullary osteosynthesis]. AB - A technique is submitted together with a device for measuring the thigh-bone proximal and distal metaepiphyses substantia spongiosa macrohardness to assess stability of the diaphyseal fractures intramedullary osteosynthesis. Data are secured on sizes of the medullary cavity cross-section, thickness of the compact bone tissue, indices for macrohardness of the femoral metaepiphyses substantia spongiosa in frontal and sagittal planes. The results obtained will, we believe, help in finding an optimum construction and technique for osteosynthesis. PMID- 11031455 TI - [Mathematical modelling and the prognosis of treatment efficacy in papillomavirus infection of the cervix uteri]. AB - In this paper, consideration is given to the problem of mathematical modelling, diagnosis and effects of treatment options on the condition of a patient exposed to papillomavirus infection. The problem is tackled of identifying the most prominent signs of degree of severity of the disease course and of therapy efficiency on the basis of parameters characterizing the immunologic vigor with making use of the covariation matrix eigenvalues algorithm, namely the modelling manifolds algorithm. Such an approach allows the central problem of classification of indices for the immunologic vigor to be settled. A mathematical model as discrimination surface to be used for prediction of results of the treatments administered is constructed. PMID- 11031456 TI - [Benign tumors of the uterus and ovaries in women with chronic obstructive bronchopulmonary diseases]. AB - Some aspects were studied of origination of uterine myoma and ovarian cyst in 121 women younger than 35 years of age presenting with chronic obstructive bronchopulmonary pathology. Uterine and ovarian benign tumors are more likely to develop against the background of chronic obstructive pulmonary pathology in the presence of respiratory failure, pulmonary emphysema, bronchiectatic disease, grave progress of the illness, development of lung disease at age, less than 10, with the above disease being more than ten years in duration. PMID- 11031457 TI - [The hypotensive effect of compression action on the chest cavity in patients with arterial hypertension]. AB - Effect were studied of a dosed chest compression on the arterial pressure (AP) in stage I-II hypertensive disease. AP was recorded at all procedures, during the whole time the respective procedure was being conducted and during five days following the course of treatment. A significant hypotensive effect was recordable in the wake of chest compression. PMID- 11031458 TI - [Seasonal fluctuations in the exocrine function of the pancreas, the state of the antioxidant system of protection and the efficacy of ethonium in chronic pancreatitis]. AB - A differentiated approach is substantiated toward treatment with aethonium (antibacterial, antiinflammatory, antioxidant drug preparation) of chronic pancreatitis with special reference to the status of lipid peroxidation, glutathionic link of antioxidant defence, and exocrine function of the pancreas (season-related rhythm of its activity), in different forms and degrees of severity included. PMID- 11031460 TI - [The catamnesis of children with celiac disease]. AB - A prospective follow-up was conducted of 29 patients with celiac disease 3 to 10 years in duration. Of these, 8 were on a spare agluten diet, in whom the time related course of clinical manifestations of the illness was characterized by stabilization of the primary symptoms of disturbed intestinal absorption, improvement in the psychoemotional state at year of their prophylactic management. The glucose tolerance test had gotten normalized after two years of being on a spare diet, lactose tolerance test remaining below the normal range in 37.5 percent of cases at year 3 of the observation, with the content of IgG antibodies to gliadine exceeding the threshold values in 33.3% on an average three times as much as the norm. In those patients having not been on a spare diet, retardation in physical development was recordable together with bone abnormalities manifested by pathologic fractures of the tubular bones. The level of anti-gliadine antibodies of both classes was found to be augmented in 71.4 percent of cases, their values having been fifty times as much as the norm. Glucose tolerance test suggested to us a steady decline in the absorption that was still to be seen at year 10 of the prospective follow-up. Lactase intolerance was somewhat less, remaining at lower values in the half of the patient population. PMID- 11031459 TI - [An unexpected stage of alkalosis in the dynamics of the early posthemorrhagic period]. AB - A study was made on acid-base metabolism in early posthemorrhagic period as exemplified by examination of patients presenting with gastrointestinal hemorrhage. It has been ascertained that hemorrhage is accompanied by a mixed variant of the acid-base state (ABS) deviation, namely metabolic lactate-acidosis and respiratory alkalosis. In the time-related course of posthemorrhagic period such deviations persist in patients with lethal outcome; with the disease running a favourable course the above deviations are found to return to normal quite soon. The development of complications leads to staging in ABC, its stages being as follows: stage I--the initial stage, stage II--persisting metabolic acidosis and respiratory alkalosis, stage III--alkalosis, stage IV--normalization, with stage III of ABS being encouraged by hypocapnia caused by function disorders of the lungs in early posthemorrhagic period, normalization of cell metabolism, increase in the rate of urination as a reflection of the third earlier identified stage of water metabolism, with the H+ excretion in the urine at the previous level. The identified ABS stage III threatens coming trouble, being accompanied by metabolic deviations together with a risk of function disorder of the myocardium. PMID- 11031461 TI - [The characteristics of secondary pneumonia (the clinical, pathogenetic and bacteriological aspects)]. AB - The analysis of 857 medical histories of patients presenting with different bodily afflictions and traumata that had come to be complicated by secondary pneumonia (SP) and of more than 2700 protocols of pathoanatomic studies made in those deceased in 35 percent of whom there had been identified morphologic changes in the lungs that presented a clinical picture of SP suggest to us that there are reasons to believe that in many instances, SP diagnosis is made difficult because of a grave condition of the patient, especially during the terminal period or the period of administering treatments when the underlying trouble symptomatology come forth rather than manifestations of pneumonia. The information secured in the pathoanatomical investigations permitted finding out that the clinical diagnosis of SP had come to be in agreement with the pathoanatomical one only in 70 percent of cases, with hypodiagnosis having come to 30%. Practitioners in their everyday work, should entertain a possibility of SP development in somatic troubles, extensive surgical interventions, traumata, wounds/injuries, burns, intoxications, and septic conditions, and they are supposed to actively prevent any possibility of origination and development of pulmonary complications. PMID- 11031462 TI - [The diagnosis and treatment of diseases of the sternum]. AB - Some aspects of diagnosis were studied together with treatment options for different disorders of the sternum, its fractures, and sternotomy-related complications in a series of 35 patients, with n = 9, 19 and 7 respectively. It has been ascertained that individualization of modalities of treatment permits securing a clinical effect in a major proportion of patients presenting with pathologies of seldom-occurring localizations. PMID- 11031463 TI - [A clinical assessment of a colorimetric method for the determination of glycosylated hemoglobin--the diabetes test]. AB - 135 patients with diabetes mellitus were studied for the level of glycated hemoglobin (GH) with the aid of the colorimetric method diabetes test involving the use of 0.3 N oxaloacetic acid, 0.06 M thiobarbituric acid, colorimetry on the spectrophotometer with the wavelength of 443 nm. No correlation was established or only weak relationship between the level of GH and chief clinical manifestations of diabetes mellitus, such as indices for metabolism, clinical presentation of the illness, degree of microangiopathy. There was no correlation between the above method and isoelectrofocusing in capillaries. Little value can be attached to GH level in controlling diabetes mellitus. PMID- 11031464 TI - [The effect of Velikobagachanskaia mineral water on the course of chronic pyelonephritis]. AB - The employing of Velikobagachanskaia mineral water internally by drinking and externally by taking a bath with it adds to the effectiveness of treatment of chronic pyelonephritis, modulates parameters associated with immune homeostasis, reduces the number of carriers of antibodies to M. hominis, facilitates prolongation of clinical and laboratory remission. PMID- 11031465 TI - [The diagnostic value of the extreme inhibition of the processes of blood lipid peroxidation]. AB - The measuring of the blood serum content of lipid peroxidation LPO products permits giving a possible prognosis of the illness. A striking decline in LPO activity is indicative of an apparent inhibition of processes of vital activity and suggests to us a possibility of a fatal outcome. PMID- 11031466 TI - [Bettolepsy and chronic pleurisy]. AB - The author submits a clinical observation of a patient who had chronic tuberculous pleurisy complicated by constriction of the root of the lung having resulted in bettolepsy. The paper contains theorization concerning its pathogenesis. An outline of differential diagnosis is given together with surgery indications. PMID- 11031467 TI - [A case of Meigs' syndrome]. AB - The described case of Meigs' [correction of Maigce] syndrome supports data on diversity of exudative pleurisies as some syndrome of manifestation of different pathological processes. Diagnosing and treatment by surgery technique of the female patient with Meigs' [correction of Magice] syndrome suggest expediency of examination by an obstetrician-gynecologist of all those women presenting with exudative pleurisy. PMID- 11031468 TI - [The clinical characteristics of progressive focal epilepsy with a herpetic etiology]. AB - Cases are described of chronic zoster encephalitis presenting with the leading progressive focal epilepsy syndrome. The clinical presentation of the trouble can include both general cerebral signs and those of the focal affection of the central nervous system only. The progressive course and duration of remission suggest to us a persisting infection manifesting against the background of immune deficiency. PMID- 11031469 TI - [The effect of Wobenzym on the atherogenic potential and inflammatory factors at the rehabilitation stage for patients who have had a myocardial infarct]. AB - Effects were studied of wobenzyme on atherogenicity of blood serum and immunoinflammatory reactions in patients with myocardial infarction during the rehabilitation stage over six months. The basis therapy when supplemented with wobenzyme was found out to facilitate normalization of the atherogenic potential and to have a positive action on the mediators of the inflammatory process. PMID- 11031471 TI - [The morphometric characteristics of the erythrocytes and their catecholamine binding properties in the treatment of stage-II hypertension patients with Renitec]. AB - Morphometric and catecholamine-binding properties of erythrocytes were studied as were their changes in monotherapy with Renitec, an inhibitor of the angiotensin converting enzyme, of patients with stage II hypertensive disease, using the image digital analysis computing system VIDAS-386 (Kontron Electronik, Germany). Patients with hypertensive disease were shown to develop degenerative changes in their erythrocytes, with catecholamine accumulation in them being a prominent feature, these events taking their course against the background of abnormalities in primary parameters characterizing cardiochemodynamics and regulation of the heart's rhythm. The above patients derived apparent benefit from treatment with Renitec, which fact was evidenced by improvement in parameters associated with the cardiochemodynamics and regulation of the cardiac rhythm, by alleviating the degree of severity of degenerative changes in erythrocytes, and by increase in their content of catecholamines. PMID- 11031470 TI - [The clinical efficacy of the calcium antagonist amlodipine in patients with ischemic heart disease and arterial hypertension]. AB - Overall, eighteen male patients with ischemic heart disease (IHD) and functional class II and III angina concurrent with stage II hypertensive disease were studied for clinical effectiveness of amlodipine (Norvasc), a new calcium antagonist, versus nifedipine and placebo. It has been ascertained that amlodipine (10 mg/day) had a significant antianginal and anti-ischemic effect and antihypertensive activity throughout 24 hours as well whereas nifedipine was effective for 6 to 8 hours. Amlodipine did not affect the well-being of the examined patients. PMID- 11031472 TI - [Systemic enzyme therapy with the preparation Wobe-Mugos E in the combined treatment of lung cancer patients]. AB - A total of 44 patients with lung carcinoma underwent combined treatment involving systemic enzyme therapy with Wobe-Mugos E. In conducting radio- and polychemotherapy, the above enzyme therapy did not complicate the postoperative course, with the pyo-septic complications having gotten reduced, the origination of pneumofibroses prevented. PMID- 11031473 TI - [The treatment results in a persistent pain syndrome in the neurological manifestation of lumbosacral osteochondrosis using osteoperiosteal electro- and laser puncture]. AB - The use of osteoperiosteal electropuncture and laser puncture combined in a complex treatment of 162 patients with lumbosacral osteochondrosis resulted in dissipation or apparent decrement in clinical manifestations of the trouble. In patients with lumboischialgia, there was an improvement in the pain syndrome as evidenced by scores on global painfulness (2.3 +/- 0.45 versus 30.2 +/- 0.62 before the treatment) in patients with degree IV pain. Less intensive pain (degree II-III as to I. P. Antonov) has gone away completely. In radicular pain syndrome, results on pain relief were much lower (P < 0.05), those on total vertebral and extravertebral painfulness much better (2.4 +/- 0.2 versus 17.3 +/- 0.7 and 1.2 +/- 0.6 versus 30.4 +/- 1.82 respectively). The secured results of treatment remained stable during two years. PMID- 11031474 TI - [The characteristics of the work process of dental technicians and their body functional status in the dynamics of work activities]. AB - The article presents in the ophthalmoergonomic aspect data from studies on the character of labour activity, condition of visual organs, vision conditions, and functions of the visual analyzer in dental mechanics. It has been ascertained that during the labour process, it is the functions of accommodation and contrast sensitivity of the eye that bear the highest strain (P < 0.05), and by the vision conditions delineation, the dental mechanic's work is on the list of those jobs requiring high visual efficiency (III B category of visual jobs according to the adopted and enforced standards classification), which facts necessitate ensuring high levels of illumination. Recommendatory measures are presented designed to prevent in dental mechanics general and visual fatigue (eyestrain and to maintain their capacity for work. PMID- 11031475 TI - [A social cultural model of the "regular period" in the sex life of men]. AB - In the experimental-testing examination of 957 patients, statistically significant results of summing up the information relevant to aspects of regular sexual intercourse in male subjects were obtained, which give good grounds for believing that the illicit/secret sexual relationship model prevailing in modern society is indeed that very social and psychological base on which further rise in venereal case rate is recordable among the population of Ukraine. And if we wish some kind of effective measures to be instituted if only to stabilize the level of venereal morbidity in the population we will certainly have to work out a comprehensive long-term state programme aimed to change motivation patterns of the social and culturological model of caring on extramarital sexual life. PMID- 11031476 TI - [The principles of the dissemination of information on sexually transmitted infections and on the drugs used in delivering medical care]. AB - An analysis is submitted of the problem of dissemination of information about infections caught by sexual intercourse or genital contact and about medication used in the treatment of this kind of pathology. The main principles are worked out of dissemination of medical information among different subjects who are interested in dissemination and securing information highlighting different aspects of the above problem. The presented approaches toward formation of information related to the problem of sexually transmitted infections permit harmonizing the interests of different subjects of the drug market and of the market of medical care to be delivered to patients presenting with the above kind of pathology. PMID- 11031477 TI - Orthopaedic proceedings 1998, 1999, 2000. PMID- 11031478 TI - [The genetic sequelae of irradiation in mammals (a review of the literature)]. AB - The problem of hereditary effects of mammal exposure to ionizing radiation has a 95-year history but to date, no simple final solution has been available. Many papers on this problem specify the dependence of the hereditary effects on dose rate, regime, physical nature of radiation exposure, type, line and age of mammals that were studied. Over many years it was studied mainly as an aspect of hereditary radiation effects in progeny of one irradiated and the second non irradiated parents. Recently due to the large-scale expansion of ionizing irradiation, it has turned out urgent to study hereditary radiation effects in progeny of both irradiated parents. However, the original studies on this problem are not numerous, and in the summarized articles, the problem practically had no specified presentation. PMID- 11031480 TI - [The effect of am umuC mutation on the induction of an SOS response in E. coli cella under the action of UV and gamma irradiation]. AB - The kinetics of the SOS induction in E. coli cells of wild type and deficient in umuC gene exposed to UV and gamma-rays were analysed. In the presence of UmuC protein SOS induction was 3-5.5 times lower and delayed for about 30 minutes after both UV and gamma rays. It was shown that decrease of the SOS induction in wild type cells irradiated by UV was due to more effective elimination of the photolesions from DNA by excision repair system. UmuCD-dependent inhibition of DNA replication was discussed as a possible mechanism allowing additional time for error-free repair. PMID- 11031479 TI - [The effect of chronic exposure to cadmium and gamma radiation at low doses on the genetic structures in mice]. AB - The DNA-protein cross-links (DPC) in mouse thymocytes and spleen lymphocytes, the number of abnormal sperm heads (ASH) and the number of micronuclei (MN) in normochromatic erythrocytes (NCE) of peripheral blood were studied in mice exposed to long-term low-intensity gamma radiation (0.072 cGy/days) and/or cadmium with drinking water (0.01 mg Cd2+/l) for 20, 40 and 80 days. The dependence of DPC level on the total dose (exposure time) of gamma radiation and/or cadmium is nonlinear. The maximal level of DPC in cells of lymphoid organs upon exposure to gamma radiation or cadmium was recorded on the 40-th day, and under combined exposure on the 20-th day of the experiment. The long-term exposure to cadmium or gamma radiation causes an increase in the ASH frequency. The increase in frequencies of MN in NCE and reciprocal translocations in spermatocytes was not found. PMID- 11031481 TI - [The characteristics of the radiation-initiated peroxidation of the phosphatidylcholine making up liposomes containing phospholipids which are susceptible to free-radical fragmentation]. AB - The regularities of accumulation of conjugated dienes and thiobarbituric acid (TBA)-reactive substances under gamma-irradiation of liposomes from rat liver phosphatidylcholine (PC) and its mixtures with the resistant to lipid peroxidation saturated phospholipids and bovine brain sphingomyelin (SM) were studied. It was established that the incorporation of negatively charged dipalmitoylphosphatidylglycerol (DPPG) and dipalmitoylphosphatidylethanol (DPPET) into lipid bilayer resulted in the increase of primary and secondary products of LPO, whereas neutral dipalmitoylphosphatidylcholine (DPPC) and SM involving in the phospholipid mixtures inhibited the peroxidation of PC. For anionic phospholipids, DPPG had more profound activating action on LPO, amongst the neutral phospholipids SM was more potent inhibitor of the reaction. Unlike DPPET and DPPC, DPPG and SM were subjected to free radical fragmentation on gamma radiation. It is suggested that the intermediates and products of free radical fragmentation may modulate the progress of LPO. PMID- 11031482 TI - [The survival of mice and the metallothionein content of their livers and kidneys as criteria for assessing exposure to pulsed neutron radiation]. AB - The effect of pulsed neutron radiation was studied in comparison with continuous neutron radiation and continuous gamma-radiation. Animal survival and induction of metallothionein (MT) synthesis in liver and kidney of mice exposed to equivalent doses were chosen as criteria for evaluation of radiation effects. It was found that the level of MT in liver and kidney of mice exposed to neutron radiation decreased 24 hours after irradiation and then continued decreasing in kidney for 48 hours after irradiation. This is evidence of more intensive free radical processes initiated by pulsed neutron radiation. At the same time, RBE values of pulsed neutrons did not differ significantly from that of continuous neutron radiation. PMID- 11031483 TI - [The role of beta-adrenoreceptors in the mechanism of the hemodynamic action and radiation-protective activity of the copolymer of 2-methyl-5-vinylpyridine with 2 methyl-5-vinylpyridinium-N-oxide]. AB - In experiment on anesthetized dogs and cats in was shown that the new water soluble copolymer initiated depressive reaction characteristic for beta adrenomimetics. This effect was levelled with the help of non-selective adrenoblockator--propranololum. In experiment on dogs the preliminary treatment with propranololum decreased the therapeutic antiradiation efficiency of the copolymer from 68.4 to 8.3%. PMID- 11031485 TI - [The dose dependence of cytogenetic damages and the adaptive response of mammalian cells under the action of ionizing radiation at low doses]. AB - The dose-effect dependence of cytogenetic damage after single dose irradiation in the dose range of 0.1-2 Gy and the adaptive response after double-dose irradiation were studied on Chinese hamster and human melanoma cells in culture. The non-linear dose dependencies were found for the induction of chromosome aberrations with decrease in cell radiosensitivity in the definite dose range. This decrease started at 10 and 20 cGy for melanoma and Chinese hamster cells respectively. The maximal adaptive response was induced at 1 cGy for melanoma cells and at 20 cGy for Chinese hamster cells. It can be supposed that the same inducible repair processes are responsible for non-linearity of dose-effect curves and induction of the adaptive response. These processes are similar in mechanisms and different in quantitative proportion for different cell types. PMID- 11031484 TI - [The accumulation of single-stranded breaks does not lead to paired DNA damage- the characteristic of the transcribing fragment of the human ribosomal operon that allows its being detected in biological fluids at the death of different body cells]. AB - It was shown by blot-hybridization with corresponding DNA probes after electrophoretic separation of control and experimental samples of human genome DNA that accumulation of single-strand breaks in the chains of double-strand fragment of transcribing range of ribosomal gene (TRrDNA) does not result in double-strand breaks. That differs from the other studied DNA sequences (cluster of histon genes, Alu-repetition, telomeric repetition and satellite III). Single strand breaks and double-strand breaks were induced by endonucleases and by gamma radiation. In spite of higher chemical modification of TRrDNA by arylazide and dimethylsulfate (because of high content of GC-pairs), under the following fragmentation TRrDNA was found to be more resistant to double-strand breaks than other studied DNA sequences. At the same time in the range of non-transcribing spacer (NTS) of ribosomal gene, the section with higher sensitivity to double strand breaks was found. Higher resistance of TRrDNA to double breaks makes it possible to identify these fragments in cell material from different tissue after death or in DNA samples after prolonged storage. Resistance of TRrDNA to formation of double-strand breaks can be used for its detection in biological fluids after cell death, including the death initiated by ionizing radiation. PMID- 11031486 TI - [The biological effects of combined exposure to low-dose irradiation and heavy metal ions]. AB - It was shown that chronic 0.25 Gy irradiation of rats (0.01 Gy/day) at background of heavy metal penetration (Cu2+, 20 mg/l or Co2+, 5 mg/l) with drinking water had caused significant accumulation of the free-radical products proportional to exhausting antioxidant and oxidizing-reduction potential in various organs and tissues. Radiation was shown to be primary harmful factor for the brain, spleen, lungs, and blood plasma, whereas the liver and heart muscle were affected with chemicals at first. Under prolonged unfavourable influence the biochemical changes observed were sustainable and progressing that satisfied to transformation of protective-adaptive reactions into prepathology stage. PMID- 11031487 TI - [The evaluation of the role of recovery from potentially lethal and sublethal damages in the RBE of densely ionizing radiation]. AB - The new data confirming the relations between RBE and recovery of cells are presented. The quantitative evaluation of the contribution of potentially lethal and sublethal damage recovery in radiosensitivity of cells of various origin after exposure to low- and high-LET ionizing radiation was carried out. The conclusion about the greater contribution of potentially lethal damage recovery in the magnitude of RBE in comparison with sublethal damage recovery was made. PMID- 11031488 TI - [Cytokines as agents for the early pathogenetic therapy of radiation injuries. Their efficacy and mechanism of action]. AB - It has been shown in experiments with three species of laboratory animals that an early administration (during the first hours following irradiation) of human recombinant interleukins 1 alpha and 1 beta separately (to mice, rats) or in combination with antibiotic therapy (dogs) substantially increases survivability, favours a more rapid regeneration of the cellular content of the bone marrow and peripheral blood, intensifies the processes of endogenous colony formation and DNA synthesis in the bone marrow and liver and lowers the expressivity of radiation-induced endotoxemia. The significance of using cytokines in the system of remedial measures in radiation pathology is discussed. PMID- 11031489 TI - The development of human exposure standards for radio-frequency fields. AB - Biological effects and health implications of radio frequency (RF) electromagnetic fields have been a subject of scientific investigation for more than 50 years. It has become a focus of attention because of the expanded use of RF radiation in the frequency range between 300 MHz and 6 GHz for wireless communication over the past decade. Another cause for the attention is the uncertainty of some observed responses and lack of understanding of the mechanism of interaction of RF electromagnetic fields with biological systems. At present, considerable efforts are devoted to developing and revising RF exposure standards. Each of these efforts should aim to make explicit the philosophy and process by which they reason and decide guidelines for deeming exposure as safe. Furthermore, the reconciliation of philosophies of protection will definitely be an asset, in practice, to those interested in international harmonization of RF exposure standards. PMID- 11031490 TI - [The effect of electromagnetic radiation on the monoamine oxidase A activity in the rat brain]. AB - The effect of the ultralow power pulse-modulated electromagnetic radiation (EMR, power density 10 microW/cm2; carrying frequency 915 MHz; modulating pulses with frequency 2, 4, 6, 8, 12, 16 and 20 Hz) on activity of monoamine oxidase (MAO-A), enzyme involved in the oxidative deamination of monoamines, was investigated. It was established that the increase of activity MAO in hypothalamus reached the maximal meaning at modulation frequency of 6 Hz that corresponded 160% (p < 0.01) of the control level; and at modulation frequency of 20 Hz the decrease of enzyme activity up to 74% (p < 0.01) was found. Mainly the action of ultralow power pulse-modulated EMR on activity of MAO in hippocamp was activating; and the maximal increase of enzyme activity up to 174% (p < 0.01) was registered at modulation frequency of 4 Hz. PMID- 11031491 TI - [A permanent magnetic field alters the phospholipase activity in the snake venom from Vipera lebetina]. AB - It was found that at the exposure of Vipera lebetina snakes (during 10 days for 30 min daily) to SMF (0.15 T1) the specific activity of venom phospholipase A1, A2 and phosphodiesterase C increased by 20.6 +/- 2.8; 31.7 +/- 3.2 and 32.7 +/- 1.3% correspondingly. The above mentioned changes of venom enzyme activity were accompanied with the decrease of its total protein amount by 31.6 +/- 2.2%. It could be supposed that the described changes are able to cause significant changes in the total metabolic activity of cells and the organism as a whole. PMID- 11031492 TI - [The action of infrasound oscillations on the properties of water and of a DNA solution]. AB - The effect of mechanical vibrations (MV) electrical conductivity of water and optical density of aqueous DNA solution. Distilled water was treated with MV of several frequencies from 3 to 5000 Hz with an intensity of 90 dB for 30 minutes. Different sensitivities of water specific electrical conductivity (SEC) were determined (the value of distilled water SEC was 209 +/- 2 microS/m). The greatest decrease of SEC (by 15.7%) under the influence of MV was observed at the frequency of 4 Hz. There was no effect at frequencies higher than 100 Hz. The treatment of DNA water solutions with MV of frequencies 4 and 10 Hz decreased its optical density by 4.2 +/- 1.1 and 4.8 +/- 1.2% correspondingly in comparison with control. In cases of treatment with frequencies of 20 and 50 Hz no effect was observed. The mechanism of MV effect on water can be connected with the changes of system structural characteristics. It is confirmed by experiments with DNA solution, where the decrease of optical density (at 260 nm) under MV treatment is conditioned with the increase of the probability of hydrogen binding formation between the bases. PMID- 11031493 TI - [An assessment of the degree of chromophore photo damage to the heme and globin components in UV-irradiated molecules and electrophoretic fractions of human carboxyhemoglobin]. AB - The contribution of hem and globin components of electrophoretic fractions of UV irradiated human carboxyhemoglobin to photodestruction of the protein was studied. The changes observed are the result of summation of some processes unequal in intensity and direction that take place in microheterogeneous media of photomodified protein. Photosensitivity of hemoproteid in electrophoretic fraction depends on apoprotein condition, whereas the hem photoresistance cannot be the evidence of the photostability of the whole molecule. PMID- 11031494 TI - [An evaluation of the biological action of increased doses of EMI in the UV range on the functional state and productivity of sheep]. AB - The effect of increased UV-radiation doses modeling 25 and 50% of ozone layer depletion on sheep's organisms was studied in the field experiment. The character of changes in animal organisms was found to depend on irradiation doses, sensitivity of individual system of living organism to electromagnetic radiation and physiological peculiarities of protection. PMID- 11031495 TI - [Individual variation of 90Sr deposition and its variability depending on the genotypic homogeneity of the sample]. AB - Individual peculiarities of 90Sr skeleton deposition in CBA mice were investigated. Differences in the radionuclide accumulation were not more than two times. Sex features in 90Sr deposition were not revealed. The range of individual variability of 90Sr deposition was analyzed in inbred mice in comparison with outbred stock and voles from natural population; it was found that variability in the radionuclide accumulation increased with sample genotypic heterogeneity. PMID- 11031496 TI - [The action of 232Th separately and in combination with heavy and alkaline metal salts on Tradescantia (clone 02)]. AB - The action of 232Th-nitrate in concentration 0.09, 0.18 and 0.36 mg/l (counting on 232Th ion) on water cultures of Tradescantia (clone 02) was investigated. It was found that all investigated concentrations of 232Th showed statistically significant genotoxic effect and increased level of morphological abnormal cells in the stamen hairs of Tradescantia in the absence of modifying action of other metal ions. Synergistic toxic interaction was found between the 232Th in concentration 0.18 mg/l and metal ions for all samples of thawed water. Synergistic genotoxic effect of the combined action of these factors was revealed only at the low total contents of ions of heavy and alkaline metals in thawed water samples. The observed synergistic effects of the combined action of 232Th and metal ions should be taken into account when controlling the radionuclide level in the environment. PMID- 11031497 TI - [An estimation of 137Cs accumulation in forest fungi by the soil properties]. AB - A method of calculation of radiocaesium uptake by forest mushrooms as a function of soil properties is presented. Calculated value of 137Cs transfer factor for Xerocomus badius in the contaminated area of Bryansk Province of Russia is in a good agreement with experimental data. PMID- 11031498 TI - [A discussion around Cohen's ecological research on the relationship between the radon content in dwellings and lung cancer mortality]. AB - B. L. Cohen showed that lung cancer lethality in U.S. sharply contradicts linear no-threshold (LNT) dependence on lung dose. This conclusion undermines established theory and practice for estimation of the irradiation remote risk; therefore, an unusual discussion has risen. Cohen's opponents (LNT adherents) said that he substituted the mean risk in a county for the individual risk that resulted in inadmissible bias of results. However, his critics' conclusions contradict many results found by Cohen and investigations by other authors. PMID- 11031499 TI - Drift-controlled anomalous diffusion: a solvable Gaussian model AB - We introduce a Langevin equation characterized by a time-dependent drift. By assuming a temporal power-law dependence of the drift, we show that a great variety of behavior is observed in the dynamics of the variance of the process. In particular, diffusive, subdiffusive, superdiffusive, and stretched exponentially diffusive processes are described by this model for specific values of the two control parameters. The model is also investigated in the presence of an external harmonic potential. We prove that the relaxation to the stationary solution has a power-law behavior in time with an exponent controlled by one of the model parameters. PMID- 11031500 TI - Fluctuation theorems for entropy production in open systems AB - We derive a fluctuation theorem to describe entropy fluctuations in steady states of systems with density gradients due to open boundaries. The fluctuations are related to the growth rate of the phase-space density, instead of the phase-space contraction rate. Explicit derivations are presented for a multibaker map, but the arguments are rather general, and should hold for a much wider class of dynamical systems. A comparison with recent results for stochastic systems is also given. PMID- 11031501 TI - Universality classes of driven lattice gases AB - Motivated by some recent criticisms to our alternative Langevin equation for driven lattice gases (DLG) under an infinitely large driving field, we revisit the derivation of such an equation, and test its validity. As a result, an additional term, coming from a careful consideration of entropic contributions, is added to the equation. This term heals all the recently reported generic infrared singularities. The emerging equation is then identical to that describing randomly driven diffusive systems. This fact confirms our claim that the infinite driving limit is singular, and that the main relevant ingredient determining the critical behavior of the DLG in this limit is the anisotropy and not the presence of a current. Different aspects of our picture are discussed, and it is concluded that it constitutes a very plausible scenario to rationalize the critical behavior of the DLG and variants of it. PMID- 11031502 TI - Driven tunneling dynamics: Bloch-Redfield theory versus path-integral approach AB - In the regime of weak bath coupling and low temperature we demonstrate numerically for the spin-boson dynamics the equivalence between two widely used but seemingly different roads of approximation, namely, the path-integral approach and the Bloch-Redfield theory. The excellent agreement between these two methods is corroborated by an efficient analytical high-frequency approach: it well approximates the decay of quantum coherence via a series of damped coherent oscillations. Moreover, a suitably tuned control field can selectively enhance or suppress quantum coherence. PMID- 11031503 TI - Detection of fixed points in spatiotemporal signals by a clustering method. AB - We present a method to determine fixed points in spatiotemporal signals. The method combines a clustering algorithm and a nonlinear analysis method fitting temporal dynamics. A 144-dimensional simulated signal, similar to a Kueppers Lortz instability, is analyzed and its fixed points are reconstructed. PMID- 11031504 TI - Experimental observation of localization in the spatial frequency domain of a kicked optical system AB - An experimental realization of an optical "kicked" system is presented. It exhibits localization analogous to that of the quantum "kicked-rotor." In the experiment, free space propagating light is periodically kicked by thin sinusoidal phase gratings, which produce high order diffractions and tend to increase the spatial frequency band. The wave property suppresses this diffusive spread. The localization is realized in a regime near anti-resonance of the system, which is also studied theoretically. The behavior in this regime is similar to that of electronic motion in incommensurate potentials. A crucial part of the experimental system is the grating in-phase positioning, which is done by using the Talbot effect. PMID- 11031505 TI - Effect of wall deformations on a confined fluid AB - We investigate the effect of wall deformations on a fluid confined in a slit of deformable walls. Monte Carlo (MC) simulations show that the fluid undergoes rarefaction or condensation depending on the wall rigidity for a small wall separation. The slope of the mean squared displacement as a function of MC steps has an algebraic dependence on the wall rigidity. The simulated density profile is qualitatively accounted for by means of a mean-field theory. PMID- 11031506 TI - Hysteresis at low Reynolds number: onset of two-dimensional vortex shedding AB - Hysteresis has been observed in a study of the transition between laminar flow and vortex shedding in a quasi-two-dimensional system. The system is a vertical, rapidly flowing soap film which is penetrated by a rod oriented perpendicular to the film plane. Our experiments show that the transition from laminar flow to a periodic von Karman vortex street can be hysteretic, i.e., vortices can survive at velocities lower than the velocity needed to generate them. PMID- 11031507 TI - Steric interaction of an incoming particle with grafted rods: exact solutions and unusual force profiles. AB - We examine the problem of a grafted rod or rods compressed by a sphere, concentrating on the steric force exerted by the rods on the sphere. We show that this problem can be solved exactly to yield simple and nontrivial expressions for the repulsive force. In particular, there are several different regimes and in some of these the force exhibits surprising maxima and minima as a function of compression. This has applications to systems of stiff grafted polymers, particularly biopolymers. One experimental realization of our system might be a rod-coil diblock copolymer with the coil grafted to a solid surface. PMID- 11031508 TI - Genuine saddle point and nucleation potential for binary systems AB - A generalized nucleation potential is constructed for binary systems. The potential consists of the reversible work of cluster formation plus additional terms arising from various kinetic effects. We show that the major nucleation flux passes through the saddle point (termed the genuine saddle point) of this generalized nucleation potential. The generalized nucleation potential reduces to the kinetic potential of a unary system when one component vanishes. The genuine saddle point concept provides a convenient way to identify systems and conditions for which the ridge crossing phenomenon occurs. Our theory agrees approximately with exact numerical results. PMID- 11031509 TI - A one- and two-dimensional nonlinear pulse interaction AB - The peculiar intergrability of the Davey-Stewartson equation allows us to analytically find solutions describing the simultaneous formation and interaction of one-dimensional and two-dimensional localized coherent structures. The predicted phenomenology allows us to address the issue of interaction of solitons of different dimensionality that may serve as a starting point for the understanding of hybrido-dimensional collisions recently observed in nonlinear optical media. PMID- 11031510 TI - Method for determining antiphase dynamics in a multimode laser AB - We measure the cross spectrum of the intensity fluctuations of pairs of modes for a multilongitudinal mode neodymium-doped yttrium aluminum garnet laser operating in the steady state regime. From the data we build up a picture of how the longitudinal mode fluctuations interfere and directly show the antiphase dynamics of the intensity fluctuations. PMID- 11031511 TI - Noncollinear generation of optical spatiotemporal solitons and application to ultrafast digital logic. AB - We demonstrate theoretically and experimentally that spatiotemporal solitons can be generated through noncollinear second-harmonic generation. The resulting Y geometry could be used to implement an optical AND gate with ultrafast, high contrast operation but without sensitivity to the phases of the input pulses. PMID- 11031512 TI - Stochastic resonance in one-dimensional diffusion with one reflecting and one absorbing end point AB - An analysis of the nonmonotonic dependence of the mean-free-passage time on the frequency of a periodic signal [stochastic resonance (SR)] for diffusion on a segment with one absorbing and one reflecting end point shows that SR exists only for some restricted values of parameters. SR always exists if the periodic telegraph signal is replaced by a random one. The latter case is considered in detail. PMID- 11031513 TI - Associative memory storing an extensive number of patterns based on a network of oscillators with distributed natural frequencies in the presence of external white noise. AB - We study associative memory based on temporal coding in which successful retrieval is realized as an entrainment in a network of simple phase oscillators with distributed natural frequencies under the influence of white noise. The memory patterns are assumed to be given by uniformly distributed random numbers on [0, 2 pi) so that the patterns encode the phase differences of the oscillators. To derive the macroscopic order parameter equations for the network with an extensive number of stored patterns, we introduce an effective transfer function by assuming a fixed-point equation of the form of the Thouless-Anderson Palmer equation, which describes the time-averaged output as a function of the effective time-averaged local field. Properties of the networks associated with synchronization phenomena for a discrete symmetric natural frequency distribution with three frequency components are studied based on the order parameter equations, and are shown to be in good agreement with the results of numerical simulations. Two types of retrieval states are found to occur with respect to the degree of synchronization, when the size of the width of the natural frequency distribution is changed. PMID- 11031514 TI - Quantum integrable system with two color components in two dimensions AB - The Davey-Stewartson 1 (DS1) system [Proc. R. Soc. London, Ser. A 338, 101 (1974)] is an integrable model in two dimensions. A quantum DS1 system with two color components in two dimensions has been reformulated. This two-dimensional problem has been reduced to two one-dimensional many-body problems with two color components. The solutions to the two-dimensional problem under consideration can be constructed from the resulting problem in one dimensions. For the latter problem with delta-function interactions and solution by the Bethe ansatz, we introduce symmetrical and antisymmetrical Young operators of the permutation group and obtain the exact solutions for the quantum DS1 system. PMID- 11031515 TI - Memory effects and macroscopic manifestation of randomness. AB - It is shown that due to memory effects the complex behavior of components in a stochastic system can be transmitted to macroscopic evolution of the system as a whole. Within the Markov approximation widely used in ordinary statistical mechanics, memory effects are neglected. As a result, a time-scale separation between the macroscopic and the microscopic level of description exists, the macroscopic differential picture is not a consequence of microscopic nondifferentiable dynamics. On the other hand, the presence of complete memory in a system means that all its components have the same behavior. If the memory function has no characteristic time scales, the correct description of the macroscopic evolution of such systems has to be in terms of the fractional calculus. PMID- 11031516 TI - Effects of random fields in an antiferromagnetic Ising spin glass AB - The effects of random fields on the two-sublattice infinite-ranged Ising spin glass model are investigated. This model is expected to be appropriate as a mean field description of antiferromagnetic spin glasses such as FexMn1-xTiO3. Within replica-symmetric calculations, we study the influence of Gaussian and bimodal random fields on the phase transitions and phase diagrams. It is shown that, in the presence of random fields, the first-order transitions are weakened and may become continuous. Also, the antiferromagnetic phases are always destroyed by sufficiently strong random fields. A qualitative comparison with existing experimental results and the limitations of the present calculations are discussed. PMID- 11031517 TI - Nose-Hoover chain method for nonequilibrium molecular dynamics simulation AB - It is demonstrated that the Nose-Hoover chain (NHC) thermostat method influences the properties of a system out of equilibrium. A simple modification of the NHC equations is proposed which enables both steady-state averages and time correlation functions to be obtained correctly for nonequilibrium states, and consistent with those computed using Gaussian isokinetic and Nose-Hoover dynamics. PMID- 11031518 TI - Performance of discrete heat engines and heat pumps in finite time AB - The performance in finite time of a discrete heat engine with internal friction is analyzed. The working fluid of the engine is composed of an ensemble of noninteracting two level systems. External work is applied by changing the external field and thus the internal energy levels. The friction induces a minimal cycle time. The power output of the engine is optimized with respect to time allocation between the contact time with the hot and cold baths as well as the adiabats. The engine's performance is also optimized with respect to the external fields. By reversing the cycle of operation a heat pump is constructed. The performance of the engine as a heat pump is also optimized. By varying the time allocation between the adiabats and the contact time with the reservoir a universal behavior can be identified. The optimal performance of the engine when the cold bath is approaching absolute zero is studied. It is found that the optimal cooling rate converges linearly to zero when the temperature approaches absolute zero. PMID- 11031519 TI - Analog of Planck's formula and effective temperature in classical statistical mechanics far from equilibrium AB - We study the statistical mechanics very far from equilibrium for a classical system of harmonic oscillators colliding with point particles (mimicking a heat reservoir), for negligible initial energies of the oscillators. It is known that for high frequencies the times of relaxation to equilibrium are extremely long, so that one meets with situations of quasiequilibrium very far from equilibrium similar to those of glassy systems. Using recent results from the theory of dynamical systems, we deduce a functional relation between energy variance and mean energy that was introduced by Einstein phenomenologically in connection with Planck's formula. It is then discussed how this leads to an analog of Planck's formula. This requires using Einstein's relation between specific heat and energy variance to define an effective temperature in a context of quasiequilibrium far from equilibrium, as is familiar for glassy systems. PMID- 11031520 TI - Analytical systematic approximate method of a two-state dissipative system AB - Occupation probability and changes of the environment in a dissipative two-state system have been investigated using a systematic approximate method. This method explores the tunneling dynamics through a system state vector with manifest physical meanings and provides a deep microscopic insight into the dynamical behaviors of the system. PMID- 11031521 TI - Fluctuation-dissipation process without a time scale AB - We study the influence of a dissipation process on diffusion dynamics triggered by fluctuations with long-range correlations. We make the assumption that the perturbation process involved is of the same kind as those recently studied numerically and theoretically, with a good agreement between theory and numerical treatment. As a result of this assumption the equilibrium distribution departs from the ordinary canonical distribution. The distribution tails are truncated, the distribution border is signaled by sharp peaks, and, in the weak dissipation limit, the central distribution body becomes identical to a truncated Levy distribution. PMID- 11031522 TI - Influence of additive noise on transitions in nonlinear systems AB - The effect of additive noise on transitions in nonlinear systems far from equilibrium is studied. It is shown that additive noise in itself can induce a hidden phase transition, which is similar to the transition induced by multiplicative noise in a nonlinear oscillator [P. Landa and A. Zaikin, Phys. Rev. E 54, 3535 (1996)]. Investigation of different nonlinear models that demonstrate phase transitions induced by multiplicative noise shows that the influence of additive noise upon such phase transitions can be crucial: additive noise can either blur such a transition or stabilize noise-induced oscillations. PMID- 11031523 TI - Diluted networks of nonlinear resistors and fractal dimensions of percolation clusters. AB - We study random networks of nonlinear resistors, which obey a generalized Ohm's law V approximately Ir. Our renormalized field theory, which thrives on an interpretation of the involved Feynman diagrams as being resistor networks themselves, is presented in detail. By considering distinct values of the nonlinearity r, we calculate several fractal dimensions characterizing percolation clusters. For the dimension associated with the red bonds we show that dred = 1/nu at least to order O(epsilon 4), with nu being the correlation length exponent, and epsilon = 6 - d, where d denotes the spatial dimension. This result agrees with a rigorous one by Coniglio. Our result for the chemical distance, dmin = 2 - epsilon/6 - [937/588 + 45/49(ln 2 - 9/10 ln 3)](epsilon/6)2 + O(epsilon 3) verifies a previous calculation by one of us. For the backbone dimension we find DB = 2 + epsilon/21 - 172 epsilon 2/9261 + 2[-74639 + 22680 zeta(3)]epsilon 3/4084101 + O(epsilon 4), where zeta(3) = 1.202057..., in agreement to second order in epsilon with a two-loop calculation by Harris and Lubensky. PMID- 11031524 TI - Nonpotential effects in dynamics of fronts between convection patterns AB - The dynamics of fronts between hexagonal convection patterns and a mechanical equilibrium state are investigated in the cases where the hexagons are stable and the equilibrium is either stable or unstable. The Newell-Whitehead-Segel, equations with additional nonpotential terms are used. The velocity of the domain boundary between two competing convection patterns and the wave number selected in the presence of the front are calculated. PMID- 11031525 TI - Detecting the onset of bifurcations and their precursors from noisy data AB - We study the problem of the detection of noise-induced precursors of periodic motion instabilities in stochastic dynamical systems. In particular, we concentrate on the period-doubling bifurcation. We have developed a statistical method to detect the onset of bifurcations and their precursors based on the previously established topological recurrence technique. PMID- 11031526 TI - Corrections to scaling in the forest-fire model. AB - We present a systematic study of corrections to scaling in the self-organized critical forest-fire model. The analysis of the steady-state condition for the density of trees allows us to pinpoint the presence of these corrections, which take the form of subdominant exponents modifying the standard finite-size scaling form. Applying an extended version of the moment analysis technique, we find the scaling region of the model and compute nontrivial corrections to scaling. PMID- 11031527 TI - Categorization in a Hopfield network trained with weighted examples: extensive number of concepts. AB - We consider the categorization problem in a Hopfield network with an extensive number of concepts p = alpha N and trained with s examples of weight lambda tau, tau = 1,...,s in the presence of synaptic noise represented by a dimensionless "temperature" T. We find that the retrieval capacity of an example with weight lambda 1, and the corresponding categorization error, depend also on the arithmetic mean lambda m of the other weights. The categorization process is similar to that in a network trained with Hebb's rule, but for lambda 1/lambda m > 1 the retrieval phase is enhanced. We present the phase diagram in the T-alpha plane, together with the de Almeida-Thouless line of instability. The phase diagrams in the alpha-s plane are discussed in the absence of synaptic noise and several values of the correlation parameter b. PMID- 11031528 TI - Wave fronts may move upstream in semiconductor superlattices AB - In weakly coupled, current biased, doped semiconductor superlattices, domain walls may move upstream against the flow of electrons. For appropriate doping values, a domain wall separating two electric-field domains moves downstream below a first critical current, it remains stationary between this value and a second critical current, and then moves upstream above. These conclusions are reached by using a comparison principle to analyze a discrete drift-diffusion model, and validated by numerical simulations. Possible experimental realizations are suggested. PMID- 11031529 TI - Disturbances in a power transmission system AB - A simple model for a power transmission system is presented. In this model, disturbances of all sizes may occur. They are randomly triggered and have the characteristic behavior of avalanches. A single parameter describes the scaling of the avalanche size. This parameter combines a measure of closeness to the maximum load, size of transferred loads during an overloading event, and connectivity of the system. The probability distribution function of the size of the disturbance has power-scaling range with the exponent close to -1. PMID- 11031530 TI - Quasistationary distributions of dissipative nonlinear quantum oscillators in strong periodic driving fields AB - The dynamics of periodically driven quantum systems coupled to a thermal environment is investigated. The interaction of the system with the external coherent driving field is taken into account exactly by making use of the Floquet picture. Treating the coupling to the environment within the Born-Markov approximation one finds a Pauli-type master equation for the diagonal elements of the reduced density matrix in the Floquet representation. The stationary solution of the latter yields a quasistationary, time-periodic density matrix which describes the long-time behavior of the system. Taking the example of a periodically driven particle in a box, the stationary solution is determined numerically for a wide range of driving amplitudes and temperatures. It is found that the quasistationary distribution differs substantially from a Boltzmann-type distribution at the temperature of the environment. For large driving fields it exhibits a plateau region describing a nearly constant population of a certain number of Floquet states. This number of Floquet states turns out to be nearly independent of the temperature. The plateau region is sharply separated from an exponential tail of the stationary distribution which expresses a canonical Boltzmann-type distribution over the mean energies of the Floquet states. These results are explained in terms of the structure of the matrix of transition rates for the dissipative quantum system. Investigating the corresponding classical, nonlinear Hamiltonian system, one finds that in the semiclassical range essential features of the quasistationary distribution can be understood from the structure of the underlying classical phase space. PMID- 11031531 TI - Tunneling through a fluctuating barrier: two-level model AB - We investigate the problem of tunneling across a randomly fluctuating barrier in the presence of dissipation in the two-level approximation. The barrier fluctuations are induced by a random telegraph noise whose switching rate nu is taken as a control parameter. For infinitely fast fluctuations the dynamics of the system is similar to the static case, while, for very small nu, the barrier evolution is a superposition of static solutions for both configurations. This leads to a resonant beating or long-time periodic localization. For an intermediate value of nu we have found a resonancelike suppression of coherent tunneling. When the system levels are detuned, a resonant enhancement of decay in the incoherent regime also occurs. PMID- 11031532 TI - Dynamics of zigzag destabilized solitary stripes in a dc-driven pattern-forming semiconductor gas-discharge system AB - Zigzag destabilization of self-organized solitary stripes was detected recently in the current density of a planar semiconductor gas discharge system. In the present work it is revealed that this instability is accompanied by the propagation of the zigzag deformation along the body of a stripe. This phenomenon is quantitatively analyzed using a high-speed image acquisition technique based on a framing camera system. The velocity of propagation has been found to increase monotonously with the global electric current, while the characteristic wavelength of the pattern shows a complicated behavior. The connection of the obtained data to available results of theoretical analysis of secondary bifurcations of solitary stripes in reaction-diffusion media is considered. PMID- 11031533 TI - Rate processes in a delayed, stochastically driven, and overdamped system AB - A Fokker-Planck formulation of systems described by stochastic delay differential equations has been recently proposed. A separation of time scales approximation allowing this Fokker-Planck equation to be simplified in the case of multistable systems is hereby introduced, and applied to a system consisting of a particle coupled to a delayed quartic potential. In that approximation, population numbers in each well obey a phenomenological rate law. The corresponding transition rate is expressed in terms of the noise variance and the steady-state probability density. The same type of expression is also obtained for the mean first passage time from a given point to another one. The steady-state probability density appearing in these formulas is determined both from simulations and from a small delay expansion. The results support the validity of the separation of time scales approximation. However, the results obtained using a numerically determined steady-state probability are more accurate than those obtained using the small delay expansion, thereby stressing the high sensitivity of the transition rate and mean first passage time to the shape of the steady-state probability density. Simulation results also indicate that the transition rate and the mean first passage time both follow Arrhenius' law when the noise variance is small, even if the delay is large. Finally, deterministic unbounded solutions are found to coexist with the bounded ones. In the presence of noise, the transition rate from bounded to unbounded solutions increases with the delay. PMID- 11031534 TI - Cluster-variation-Pade-approximant method for the simple cubic Ising model AB - The cluster-variation-Pade-approximant method is a recently proposed tool, based on the extrapolation of low- and high-temperature results obtained with the cluster-variation method, for the determination of critical parameters in Ising like models. Here the method is applied to the three-dimensional simple cubic Ising model, and new results, obtained with an 18-site basic cluster, are reported. Other techniques for extracting nonclassical critical exponents are also applied and their results compared with those by the cluster-variation-Pade approximant method. PMID- 11031535 TI - Invasion percolation with long-range correlations: first-order phase transition and nonuniversal scaling properties AB - We present the results of extensive Monte Carlo simulations of the invasion percolation model with trapping (TIP) with long-range correlations, a problem which is relevant to multiphase flow in field-scale porous media, such as oil reservoirs and groundwater aquifers, as well as flow in rock fractures. The correlations are generated by a fractional Brownian motion characterized by a Hurst exponent H. We employ a highly efficient algorithm for simulating TIP, and a novel method for identifying the backbone of TIP clusters. Both site and bond TIP are studied. Our study indicates that the backbone of bond TIP is loopless and completely different from that of site TIP. We obtain precise estimates for the fractal dimensions of the sample-spanning cluster (SSC), the minimal path, and the backbone of site and bond TIP, and analyze the size distribution of the trapped clusters, in order to identify all the possible universality classes of TIP with long-range correlations. For site TIP with H > 1/2 the SSC and its backbone are compact, indicating a first-order phase transition at the percolation threshold, while the minimal paths are essentially straigth lines. For H < 1/2 the SSC, its backbone, and the minimal paths are all fractal with fractal dimensions that depend on the Hurst exponent H. The fractal dimension of the loopless backbone for bond TIP is much less than that of site TIP for any H. PMID- 11031536 TI - Crossover from nonclassical to classical chemical kinetics in an initially separated A + B<-->C reaction-diffusion system with arbitrary diffusion constants. AB - The asymptotic long-time properties of the reaction front formed in a reversible reaction-diffusion process A + B<-->C with initially separated reactants are investigated. The case of arbitrary nonzero values of the diffusion constants DA, DB, DC of the components A, B, C and the initial concentrations a0 and b0 of A and B is considered. The system is studied in the limit of g-->0, where g is the backward reaction rate constant. In accordance with previous work, the dynamics of the reaction front is described as a crossover between the "irreversible" regime at times t << g-1 and the "reversible" regime at times t >> g-1. It is shown that through this crossover the macroscopic properties of the reaction front, such as the global rate of C production, the motion of the reaction zone center, and the concentration profiles of the components outside the reaction front, are unchanged. The concentration profiles of the components inside the reaction zone are described by quasistatic equations. The results of the theoretical consideration are confirmed by computing the mean-field kinetics equations. PMID- 11031537 TI - Spiral wave dynamics in oscillatory inhomogeneous media. AB - The effect of a long length scale static inhomogeneity on spiral wave dynamics is studied in the two-dimensional complex Ginzburg-Landau equation. We find that the inhomogeneity leads to the formation of a dominant spiral domain that suppresses other spiral domains, and that the spiral vortices slowly drift in the presence of an inhomogeneity with a velocity that is proportional to the local parameter gradients. We derive an expression for the spiral vortex drift velocity and present examples of both fixed point and limit cycle attractors of the spiral vortices. PMID- 11031538 TI - Noise sustained propagation: local versus global noise AB - We expand on prior results on noise supported signal propagation in arrays of coupled bistable elements. We present and compare experimental and numerical results for kink propagation under the influence of local and global fluctuations. As demonstrated previously for local noise, an optimum range of global noise power exists for which the medium acts as a reliable transmission "channel." We discuss implications for propagation failure in a model of cardiac tissue, and present a general theoretical framework based on discrete kink statistics. Valid for generic bistable chains, the theory captures the essential features observed in our experiments and numerical simulations. PMID- 11031539 TI - Unequal intralayer coupling in a bilayer driven lattice gas AB - The system under study is a twin-layered lattice gas at half filling, being driven to nonequilibrium steady states by a large, finite "electric" field. By making intralayer couplings unequal, we were able to extend the phase diagram obtained by Hill, Zia, and Schmittmann and found some interesting effects. Many transient phases to the strip phase were found to be long lived. We also attempted to test whether the driven lattice gas with negative interlayer coupling is still in the Ising universality class. Simulation results suggest a value of 1.75 for the exponent gamma but a value closer to 2.0 for the ratio gamma/nu. We suspect a different susceptibility has to be used due to the presence of two phases near criticality. PMID- 11031540 TI - Impact of environmental noise on oscillatory pattern formation in crystal growth: plagioclase feldspar AB - Recently, a model for oscillatory zoning in a geochemical system (plagioclase feldspar) was proposed. In that model, oscillations in the composition are generated through a Hopf bifurcation. In this paper, the effects of fluctuations of the bulk composition are studied by means of computer simulations. It is shown that environmental noise can lead to pattern formation such as oscillatory zoning, even when no deterministic periodic solutions exist. The fluctuations in the bulk composition thus lead to an enlargement of the range of system's variables values for which oscillatory zoning occurs. Coherence resonance close to the Hopf bifurcation is also observed in such a system. PMID- 11031541 TI - One-dimensional partially asymmetric simple exclusion process on a ring with a defect particle AB - The effect of a moving defect particle for the one-dimensional partially asymmetric simple exclusion process on a ring is considered. The current of the ordinary particles, the speed of the defect particle, and the density profile of the ordinary particles are calculated exactly. The phase diagram for the correlation length is identified. As a by-product, the average and the variance of the particle density of the one-dimensional partially asymmetric simple exclusion process with open boundaries are also computed. PMID- 11031542 TI - Integrated approach to the assessment of long range correlation in time series data AB - To assess whether a given time series can be modeled by a stochastic process possessing long range correlation, one usually applies one of two types of analysis methods: the spectral method and the random walk analysis. The first objective of this work is to show that each one of these methods used alone can be susceptible to producing false results. We thus advocate an integrated approach which requires the use of both methods in a consistent fashion. We provide the theoretical foundation of this approach and illustrate the main ideas using examples. The second objective relates to the observation of long range anticorrelation (Hurst exponent H < 1/2) in real world time series data. The very peculiar nature of such processes is emphasized in light of the stringent condition under which such processes can occur. Using examples, we discuss the possible factors that could contribute to the false claim of long range anticorrelations, and demonstrate the particular importance of the integrated approach in this case. PMID- 11031543 TI - Percolation and spatial correlations in a two-dimensional continuum deposition model AB - We introduce a two-dimensional continuum deposition model of spatially extended objects, with an effective repulsive contact interaction between them represented by a parameter 0 < or = q < or = 1. For q = 0, the deposited network is uniformly random, while for q = 1 particles are not allowed to overlap. For 0 < or = q < 1, we carry out extensive simulations on fibers, needles, and disks to study the dependence of the percolation threshold on q. We derive expressions for the threshold near q = 0 and q = 1 and find good qualitative agreement with the simulations. The deposited networks produced by the model display nontrivial density correlations near percolation threshold. These are reflected in the appropriate spatial correlation functions. We study such functions close to q = 1 and derive an approximate expression for the pair distribution function. PMID- 11031544 TI - Effect of bulk magnetic field on critical Ising films AB - Two-dimensional Ising films L x infinity in a nonvanishing bulk magnetic field H are studied at the bulk critical temperature Tc for two choices of surface fields (a) H1 = HL = 0 (ordinary transition), and (b) H1 = HL = infinity (normal transition) by the density-matrix renormalization-group method. Universal scaling functions for magnetization profiles, the excess magnetization gamma, the longitudinal correlation length xi parallel, and for the analog of the solvation force fsolv are found and discussed. When H1 = 0 the scaling function for fsolv has two symmetric minima at y = sgn(H)L magnitude of H nu/delta approximately +/- 1 with an amplitude at the minimum about 3.8 times the value at H = 0, the Casimir amplitude. For the normal transition the scaling function for fsolv has a single minimum near the continuation of the pseudocoexistence (capillary condensation) line, with an amplitude about 100 times the Casimir amplitude. PMID- 11031545 TI - Topological scaling and gap filling at crisis AB - Scaling laws associated with an interior crisis of chaotic dynamical systems are studied. We argue that open gaps of the chaotic set become densely filled at the crisis due to the sudden appearance of unstable periodic orbits with extremely long periods. We formulate a scaling theory for the associated growth of the topological entropy. PMID- 11031546 TI - Extracting dynamics from threshold-crossing interspike intervals: possibilities and limitations. AB - In this paper we estimate dynamical characteristics of chaotic attractors from sequences of threshold-crossing interspike intervals, and study how the choice of the threshold level (which sets the equation of a secant plane) influences the results of the numerical computations. Under quite general conditions we show that the largest Lyapunov exponent can be estimated from a series of return times to the secant plane, even in the case when some of the loops of the phase space trajectory fail to cross this plane. PMID- 11031547 TI - Influence of stable Floquet exponents on time-delayed feedback control AB - The performance of time-delayed feedback control is studied by linear stability analysis. Analytical approximations for the resulting eigenvalue spectrum are proposed. Our investigations demonstrate that eigenbranches that develop from the stable Lyapunov exponents of the free system also have a strong influence on the control properties, either by hybridization or by a crossing of branches which interchanges the role of the leading eigenvalue. Our findings are confirmed by numerical analysis of two particular examples, the Toda and the Rossler models. More important is the verification by actual electronic circuit experiments. Here, the observed reduction of control domains can be attributed to these additional eigenvalue branches. The investigations lead to a thorough analytical understanding of the stability properties in time-delayed feedback systems. PMID- 11031548 TI - Quantum resonances and regularity islands in quantum maps AB - We study analytically as well as numerically the dynamics of a quantum map near a quantum resonance of an order q. The map is embedded into a continuous unitary transformation generated by a time-independent quasi-Hamiltonian. Such a Hamiltonian generates at the very point of the resonance a local gauge transformation described by the unitary unimodular group SU(q). The resonant energy growth is attributed to the zero Liouville eigenmodes of the generator in the adjoint representation of the group while the nonzero modes yield saturating with time contribution. In a vicinity of a given resonance, the quasi-Hamiltonian is then found in the form of power expansion with respect to the detuning from the resonance. The problem is related in this way to the motion along a circle in a (q2 - 1)-component inhomogeneous "magnetic" field of a quantum particle with q intrinsic degrees of freedom described by the SU(q) group. This motion is in parallel with the classical phase oscillations near a nonlinear resonance. The most important role is played by the resonances with the orders much smaller than the typical localization length q << l. Such resonances master for exponentially long though finite times the motion in some domains around them. Explicit analytical solution is possible for a few lowest and strongest resonances. PMID- 11031549 TI - Semiclassical inequivalence of polygonalized billiards AB - Polygonalization of any smooth billiard boundary can be carried out in several ways. We show here that the semiclassical description depends on the polygonalization process and the results can be inequivalent. We also establish that generalized tangent polygons are closest to the corresponding smooth billiard and for de Broglie wavelengths larger than the average length of the edges, the two are semiclassically equivalent. PMID- 11031550 TI - Three coupled oscillators as a universal probe of synchronization stability in coupled oscillator arrays AB - We show that the stability surface that governs the synchronization of a large class of arrays of identical oscillators can be probed with a simple array of just three identical oscillators. Experimentally this implies that it may be possible to probe the synchronization conditions of many arrays all at the same time. In the process of developing a theory of the three-oscillator probe, we also show that several regimes of asymptotic coupling can be derived for the array classes, including the case of large imaginary coupling, which apparently has not been explored. PMID- 11031551 TI - Learning-induced synchronization of a globally coupled excitable map system. AB - We propose a pulse-coupled neural network model in which one-dimensional excitable maps connected in a time-delayed network serve as the neural processing units. Although the individual processing unit has simple dynamical properties, the network exhibits collective chaos in the active states. Introducing a Hebbian learning algorithm for synaptic connections enhances the synchronization of excitation timing of the units within a subpopulation. The synchronizing clusters approximately exhibit a power-law size distribution, suggesting a hierarchy of synchronization. After applying a stationary signal to a subpopulation of the units with learning, the network then reproduces the signal. The learnable time range is much longer than the inherent time scale of the processing units, i.e., the synaptic delay time. Also, the network can reproduce periodic signals with time resolution finer than the delay time. Our present network model can be considered as a temporal association device which operates in chaotic states. PMID- 11031552 TI - Shape of attractors for three-dimensional dissipative dynamical systems AB - We introduce a method to bound attractors of dissipative dynamical systems in phase and parameter spaces. The method is based on the determination of families of transversal surfaces (surfaces crossed by the flow in only one direction). This technique yields very restrictive geometric bounds in phase space for the attractors. It also gives ranges of parameters of the system for which no chaotic behavior is possible. We illustrate our method on different three-dimensional dissipative systems. PMID- 11031553 TI - Classical limit in terms of symbolic dynamics for the quantum baker's map AB - We derive a simple closed form for the matrix elements of the quantum baker's map that shows that the map is an approximate shift in a symbolic representation based on discrete phase space. We use this result to give a formal proof that the quantum baker's map approaches a classical Bernoulli shift in the limit of a small effective Planck's constant. PMID- 11031554 TI - Anticipating chaotic synchronization AB - Dissipative chaotic systems with a time-delayed feedback can drive near-identical systems in such a way that the driven systems anticipate the drivers by synchronizing with their (arbitrarily distant) future states. This counterintuitive behavior is globally stable, robust, and a pure result of the interplay between delayed feedback and dissipation. Thus it constitutes a rather universal phenomenon of nonlinear dynamics. For small anticipation times, anticipating synchronization also occurs in chaotic systems without a memory term in the driver. PMID- 11031555 TI - Eliminating spatiotemporal chaos and spiral waves by weak spatial perturbations AB - The possibility of eliminating spatiotemporal chaos and spiral waves by weak spatial perturbations in a spatially extended dynamical system is demonstrated numerically through the example of a wide-aperture laser. The time-independent weak spatial perturbation can effectively migrate the system from the state of spatiotemporal chaos or spiral waves to that of traveling waves. The threshold and the controllable range of the control parameters are given. By varying the amplitude or the spatial wave vector of the perturbation, drastic changes in the spatiotemporal dynamics are found. PMID- 11031556 TI - Dynamical localization and partial-barrier localization in the Paul trap AB - We investigate the nature of quantum localization exhibited by the center-of-mass motion of an ion in a Paul trap interacting with a standing laser field. Depending upon system parameters and the initial location of the ion, quantum suppression of chaotic diffusion is dominated by dynamical localization or localization due to partial barriers formed of broken separatrices and cantori. PMID- 11031557 TI - Closed almost-periodic orbits in semiclassical quantization of generic polygons AB - Periodic orbits are the central ingredients of modern semiclassical theories and corrections to these are generally nonclassical in origin. We show here that, for the class of generic polygonal billiards, the corrections are predominantly classical in origin owing to the contributions from closed almost-periodic (CAP) orbit families. Furthermore, CAP orbit families outnumber periodic families but have comparable weights. They are hence indispensable for semiclassical quantization. PMID- 11031558 TI - Stochastic resonance in noisy maps as dynamical threshold-crossing systems AB - Interplay of noise and periodic modulation of system parameters for the logistic map in the region after the first bifurcation and for the kicked spin model with Ising anisotropy and damping is considered. For both maps two distinct symmetric states are present that correspond to different phases of the period-2 orbit of the logistic map and to disjoint attractors of the spin map. The periodic force modulates the transition probabilities from any state to the opposite one symmetrically. It follows that the maps behave as threshold-crossing systems with internal dynamics, and stochastic resonance (maximum of the signal-to-noise ratio in the signal reflecting the occurrence of jumps between the symmetric states) in both models is observed. Numerical simulations agree qualitatively with analytic results based on the adiabatic theory. PMID- 11031559 TI - Learning driver-response relationships from synchronization patterns. AB - We test recent claims that causal (driver-response) relationships can be deduced from interdependencies between simultaneously measured time series. We apply two recently proposed interdependence measures that should give results similar to cross predictabilities used by previous authors. The systems that we study are asymmetrically coupled simple models (Lorenz, Roessler, and Henon models), the couplings being such that they lead to generalized synchronization. If the data were perfect (noise-free, infinitely long), we should be able to detect, at least in some cases, which of the coupled systems is the driver and which the response. This might no longer be true if the time series has finite length. Instead, estimated interdependencies depend strongly on which of the systems has a higher effective dimension at the typical neighborhood sizes used to estimate them, and causal relationships are more difficult to detect. We also show that slightly different variants of the interdependence measure can have quite different sensitivities. PMID- 11031560 TI - Adaptive estimation and control method for unstable periodic dynamics in spike trains. AB - Dynamical control of excitable biological systems is often complicated by the difficult and unreliable task of precontrol identification of unstable periodic orbits (UPO's). Here we show that, for both chaotic and nonchaotic systems, UPO's can be located, and their dynamics characterized, during control. Tracking of system nonstationarities emerges naturally from this approach. Such a method is potentially valuable for the control of spike trains of excitable biological systems, for which precontrol UPO identification is often impractical, and nonstationarities (natural or stimulation induced) are common. PMID- 11031562 TI - Noise-induced chaos in an optically injected semiconductor laser model AB - The chaos induced by an intrinsic spontaneous-emission noise in an optically injected semiconductor laser is investigated through a single-mode injection model. A method is developed to quantitatively study the scale-dependent noise effect in general, and the noise-induced chaotic feature in particular. We find that noise at an experimentally measured level can induce chaos in the system. This suggests that noise-induced chaos may indeed exist in real systems. Certain required characteristics for noise to induce chaos are identified: the periodic state itself, when subject to weak noise, should undergo a process that is much more diffusive than the Brownian motion, and the adjacent chaotic states should still behave chaotically on certain finite scales when subject to noise. We believe they are generic features for noise to induce chaos. The correlation dimension of the clean and noisy attractors is also calculated to study noise induced changes in the geometrical structure of the attractors. PMID- 11031563 TI - Hamiltonian dynamics and geometry of phase transitions in classical XY models AB - The Hamiltonian dynamics associated with classical, planar, Heisenberg XY models is investigated for two- and three-dimensional lattices. In addition to the conventional signatures of phase transitions, here obtained through time averages of thermodynamical observables in place of ensemble averages, qualitatively different information is derived from the temperature dependence of Lyapunov exponents. A Riemannian geometrization of Newtonian dynamics suggests consideration of other observables of geometric meaning tightly related to the largest Lyapunov exponent. The numerical computation of these observables- unusual in the study of phase transitions--sheds light on the microscopic dynamical counterpart of thermodynamics, also pointing to the existence of some major change in the geometry of the mechanical manifolds at the thermodynamical transition. Through the microcanonical definition of the entropy, a relationship between thermodynamics and the extrinsic geometry of the constant energy surfaces sigma E of phase space can be naturally established. In this framework, an approximate formula is worked out determining a highly nontrivial relationship between temperature and topology of sigma E. From this it can be understood that the appearance of a phase transition must be tightly related to a suitable major topology change of sigma E. This contributes to the understanding of the origin of phase transitions in the microcanonical ensemble. PMID- 11031561 TI - Synchronization plateaus in a lattice of coupled sine-circle maps. AB - Frequency synchronization is a common phenomenon in spatially extended dynamical systems, like oscillator chains and coupled map lattices. We study the distribution of synchronization plateaus in a sine-circle map lattice with a variable range coupling and randomly distributed natural frequencies. A transition between synchronized and nonsynchronized phases is observed as the coupling range is varied. The lengths of the synchronization plateaus are found to obey an exponential distribution for local coupling. PMID- 11031564 TI - Onset of convection in a binary mixture near the plait point AB - Recent experiments on the onset of convection near the liquid-vapor critical point of 3He have shown the crossover from the Rayleigh criterion to the Schwarzschild criterion for the threshold of convection as the critical point is approached. In contrast we show that for 3He-4He mixtures near the liquid-vapor critical point (plait point), the Rayleigh criterion would hold right through. Interestingly enough, this is a consequence of the proper boundary conditions. This prediction should be easy to test experimentally. PMID- 11031565 TI - Multiscale correlations and conditional averages in numerical turbulence AB - The equations of motion for the nth order velocity differences raise the interest in correlation functions containing both large and small scales simultaneously. We consider the scaling of such objects and also their conditional average representation with emphasis on the question of whether they behave differently in the inertial or the viscous subranges. The turbulent flow data are obtained by Navier-Stokes solutions on a 60(3) grid with periodic boundary conditions and Re lambda = 70. Our results complement previous high Re data analysis based on measured data [A. L. Fairhall, V. S. L'vov, and I. Procaccia, Europhys. Lett 43, 277 (1998)] whose preference were the larger scales, and the analysis of both experimental and synthetic turbulence data by [R. Benzi and co-workers, Phys. Rev. Lett. 80, 3244 (1998); Phys. Fluids 11, 2215 (1999)]. The inertial range fusion rule is confirmed and insight is obtained for the conditional averages (the local dissipation rate conditioned on the velocity fluctuations). PMID- 11031566 TI - Electromotive force for an anisotropic turbulence: intermediate nonlinearity AB - A nonlinear electromotive force for an anisotropic turbulence in the case of intermediate nonlinearity is derived. The intermediate nonlinearity implies that the mean magnetic field is not strong enough to affect the correlation time of a turbulent velocity field. The nonlinear mean-field dependencies of the hydrodynamic and magnetic parts of the alpha effect, turbulent diffusion, and turbulent diamagnetic and paramagnetic velocities for an anisotropic turbulence are found. It is shown that the nonlinear turbulent diamagnetic and paramagnetic velocities are determined by both an inhomogeneity of the turbulence and an inhomogeneity of the mean magnetic field B. The latter implies that there are additional terms in the turbulent diamagnetic and paramagnetic velocities variation of [symbol: see text]B2 and variation of (B.[symbol: see text])B. These effects are caused by a tangling of a nonuniform mean magnetic field by hydrodynamic fluctuations. This increases the inhomogeneity of the mean magnetic field. It is also shown that in an isotropic turbulence the mean magnetic field causes an anisotropy of the nonlinear turbulent diffusion. Two types of nonlinearities in magnetic dynamo determined by algebraic and differential equations are discussed. Nonlinear systems of equations for axisymmetric alpha omega dynamos in both spherical and cylindrical coordinates are derived. PMID- 11031567 TI - Motion of hydrogen bonds in diluted HDO/D2O solutions: direct probing with 150 fs resolution AB - An experiment is described to study temporal variations of the hydrogen bond length in diluted HDO/D2O solutions. The principles of this laser spectroscopic experiment are explained first. The construction of a laser source generating 150 fs pulses in the 2.5-4.5 microns spectral region at a 10 microJ power level is detailed next. The OH stretching band is reproduced for different excitation frequencies and different pump-probe delay times. A theory, based on statistical mechanics of nonlinear optical processes, is proposed to calculate the lowest two spectral moments. An effect is reported, the delay dependent vibrational solvatochromism. It is shown how this effect can be exploited to follow temporal variations of the OH...O bond length directly, in real time. The corresponding time scales are of the order of 700 fs. No bond oscillations are observed. PMID- 11031568 TI - Density-functional theory for vacancies in hard-sphere crystals AB - The equilibrium vacancy concentration in solids can be computed from density functional theory (DFT) if allowance is made for density profiles with less than one particle per lattice site. For the fundamental-measure theory (FMT), this approach predicts reasonably small vacancy concentrations in hard sphere crystals, in contrast to earlier DFTs. Using an asymptotic analysis of the FMT functional, it is shown that the number of vacancies depends exponentially on the distance to the close packing density, as expected from heuristic arguments. The prefactor of the exponential is calculated for three recently suggested variants of the theory, using density profiles obtained from a quasifree minimization. Extrapolation of the asymptotic behavior to the melting density yields good agreement with other estimates and computer simulation results. PMID- 11031569 TI - Influence of vacancies on the melting transition of hard disks in two dimensions AB - We present the results of molecular dynamics simulations of two-dimensional (2D) hard disk systems in the vicinity of melting. The simulations are used to calculate the elastic constants, which can be used to estimate the location of the Kosterlitz-Thouless dislocation unbinding transition. Simulations on defect free lattices indicate that this transition is expected to occur at essentially the same density as a first-order solid-isotropic transition and so it is not possible to rule out either a one step weak first-order transition between the solid and the isotropic fluid or a two step transition via a hexatic phase. Simulations performed on systems with vacancies indicate that the elastic constants are essentially unchanged at constant density. This result implies that vacancies have little influence on the melting of 2D hard disk solids. PMID- 11031570 TI - Dissipation field asymmetry and intermittency in fully developed turbulence AB - Experimental study of high Reynolds number turbulence provides additional evidence that asymmetry of turbulence is related to the intermittency. The refined similarity hypothesis (RSH), on the other hand, connects the intermittency of the longitudinal velocity increments with that of the dissipation field, implying in particular that the dissipation field should be asymmetric as well. The asymmetry of the latter is indeed found in these experiments. In addition, the study of the dissipation field asymmetry provides us with quantitative estimations of the deviations from the RSH. PMID- 11031571 TI - Transition to turbulent thermal convection beyond Ra = 10(10) detected in numerical simulations AB - We have conducted high-resolution two-dimensional calculations for a Boussinesq convection model with a Prandtl number of unity in an aspect-ratio 3 box, going from Rayleigh numbers between 10(8) to 10(14). A grid of 1024 x 3076 grid points consisting of a cosine-sine basis set has been employed for free-slip boundary conditions. We have found evidence for a transition involving the branching of plumes at a Rayleigh number of 10(10). Inside the core of these "superplumes," the structure is extremely complex. There may be another transition at Ra of 10(12), where a secondary instability may develop in regions of the local Rayleigh number which becomes supercritical inside the core of the complex "superplumes." For Ra of 10(8) to 10(10), Ra follows a 1/3 power law in the Nusselt-Rayleigh number relationship. From Ra of 10(10) to 10(12), Ra follows a 1/2 power law. Above this value the Nusselt number becomes insensitive to the variation in the global Rayleigh number and this is due to the development of small-scale convection cells vertically aligned in the interior of the extremely high Ra number flow. The global Reynolds number scales as Re approximately Ra1/4 up to Ra of 10(14). Scaling relationships based on global properties would not work in extremely high Ra situations beyond Ra of 10(12) because of the complex turbulent layered convection in the core of the flow and the severe degradation of the boundary layers. PMID- 11031572 TI - Bifurcation theory for three-dimensional flow in the wake of a circular cylinder AB - A bifurcation scenario is presented for three-dimensional vortex shedding in the wake of a circular cylinder for Reynolds numbers up to 300. Amplitude equations are proposed to describe the nonlinear interaction between two three-dimensional modes of shedding with different spanwise wave numbers and different spatiotemporal symmetries. The amplitude equations explain many features of the transition scenario observed experimentally. PMID- 11031573 TI - Mie scattering from a sonoluminescing bubble with high spatial and temporal resolution AB - The dynamics of a single-air bubble trapped in a resonant sound field in water has been characterized by Mie scattering and a Streak camera with high spatial and temporal resolution. The streak images show that in the endphase of the cavitation collapse the scattered light intensity is no function of the bubble radius anymore. In the last nanoseconds around minimum bubble radius most of the light is scattered at the highly compressed water surrounding the bubble and not at the bubble wall. This leads to a minimum in the scattered light intensity about 700 ps before the sonoluminescence pulse is emitted. And neglecting this changes leads to a strong overestimation of the bubble-wall velocity. In the reexpansion phase the high spatial resolution of the streak camera allows one to distinguish between the light scattered at the bubble wall and the light scattered at the outgoing shock wave. PMID- 11031574 TI - Contact line dynamics near the pinning threshold: a capillary rise and fall experiment AB - We used video microscopy to study the pinning dynamics of air/water contact lines in vertical glass capillaries. Stick-slip behavior and avalanches are observed in tubes with rough interior walls and strong pinning forces. In tubes with smooth interior walls, we find that receding contact lines in falling water columns show no evidence of pinning, but advancing contact lines in rising water columns exhibit algebraic slow down. The measured value of the critical exponent beta varies from run to run, but it is always larger than unity. Furthermore, we find that the rise dynamics varies with the waiting time preceding the experiments. These observations led us to conclude that the wetting film on the surface and other microscopic changes in the slipping region near the contact line affect the macroscopic dynamics. We discuss the differences between the real system and the existing theories that might explain the results. We also present a brief review of other studies of contact line dynamics and a numerical study of a one dimensional model. PMID- 11031575 TI - Thermal noise and the stability of single sonoluminescing bubbles AB - The stability of a bubble levitated in an acoustic field under single bubble sonoluminescence conditions was numerically investigated taking thermal noise effects into consideration. Due to the microscopic size of a sonoluminescing bubble thermal noise is important to its surface and is found to cause small irregularities in its spherical shape. A stochastic differential equation in Langevin form is derived to describe the dynamics of a perturbation from the spherical and solved together with the Rayleigh-Plesset equation. The mechanisms responsible for the amplification of small irregularities are examined and a stability threshold is derived, which is in good agreement with experimental threshold data of Holt and Gaitan. PMID- 11031576 TI - Toward a self-generating magnetic dynamo: the role of turbulence AB - Turbulent flow of liquid sodium is driven toward the transition to self generating magnetic fields. The approach toward the transition is monitored with decay measurements of pulsed magnetic fields. These measurements show significant fluctuations due to the underlying turbulent fluid flow field. This paper presents experimental characterizations of the fluctuations in the decay rates and induced magnetic fields. These fluctuations imply that the transition to self generation, which should occur at larger magnetic Reynolds number, will exhibit intermittent bursts of magnetic fields. PMID- 11031577 TI - Lattice-Boltzmann algorithm for simulating thermal two-phase flow AB - An algorithm is described for incorporating thermal effects into lattice Boltzmann simulations of two-phase flow. This algorithm is a combination of a two distribution model for simulating single-phase thermal flow recently proposed by the authors and the thermodynamically-based model for isothermal two-phase flow of Swift et al. [Phys. Rev. E 54, 5041 (1996)]. The algorithm also corrects a problem with the original single-phase thermal flow model, which described the thermal energy flux as proportional to a gradient of the internal energy instead of being proportional to the gradient of the temperature. For ideal-gas systems, these two descriptions are equivalent but for nonideal systems there is a systematic discrepancy between the original thermal model and classical hydrodynamics. The algorithm is tested on several simple problems. These include formation of a free-standing isothermal thin liquid film, evaporation of a thin liquid film from a heated plate, evaporation of an isolated droplet, and condensation of liquid in a channel. Where possible, the simulations are compared against known analytic results. PMID- 11031578 TI - Gravity in a lattice Boltzmann model AB - In this paper we consider the introduction of a body force, in the incompressible limit, into the lattice Boltzmann model. A number of methods are considered and their suitability to our objectives determined. When there is no density variation across the fluid, gravity can be introduced in the form of an altered pressure gradient. This method correctly satisfies the Navier-Stokes equation; however, if there is a non-negligible density variation present (produced by the body force or otherwise) this method becomes less accurate as the density variation increases and the constant density approximation becomes less valid. Three other methods are also considered for application when there is a non negligible density variation. The equations of motion satisfied by these models are found up to second order in the Knudsen number and it is seen that only one of these methods satisfies the true Navier-Stokes equation. Numerical simulations are performed to compare the different models and to assess the range of application of each. PMID- 11031579 TI - Exact relationship for third-order structure functions in helical flows AB - An exact law for turbulent flows is written for third-order structure functions taking into account the invariance of helicity, a law akin to the so-called "4/5 law" of Kolmogorov. Here, the flow is assumed to be homogeneous, incompressible and isotropic but not invariant under reflectional symmetry. Our result is consistent with the derivation by O. Chkhetiani [JETP Lett. 10, 808, (1996)] of the von Karman-Howarth equation in the helical case, leading to a linear scaling relation for the third-order velocity correlation function. The alternative relation of the Kolmogorov type we derive here is written in terms of mixed structure functions involving combinations of differences of all components for both the velocity and vorticity fields. This relationship could prove to be a stringent test for the measuring of vorticity in the laboratory, and provide a supplementary tool for the study of the properties of helical flows. PMID- 11031580 TI - Bromomalonic-acid-induced transition from trigger wave to big wave in the Belousov-Zhabotinsky reaction AB - The Marangoni effects on chemical waves in the ferroin-catalyzed Belousov Zhabotinsky reaction were studied. The main purpose of the present study was to understand the mechanism of the big wave, an accelerative chemical wave involving surface-tension-driven fluid motions. Spatiotemporal variations of surface tension caused by a chemical wave were measured using the Wilhelmy method. The transition from conventional trigger waves to big waves, due to a concentration change of bromomalonic acid, was observed. The strong surface activity of the bromomalonic acid which was responsible for the transition was also observed. It led to an acceleration of the big waves through the Marangoni effect. PMID- 11031581 TI - Structure and thermodynamic properties of a binary liquid in a porous matrix: the formalism AB - Using the replica trick we derive a formalism to describe the structure and the thermodynamic properties of a binary liquid in equilibrium with a porous medium. We present the replica Ornstein-Zernike equations for the general case of a k component liquid inside a porous matrix; besides the usual liquid-state closure relations, we consider in particular the optimized random phase approximation (ORPA) restricting ourselves at present to hard-core potentials exclusively. We present furthermore several thermodynamic relations: the Gibbs-Duhem equation, the compressibility, and the viral equation. Within the framework of the ORPA (mean spherical approximation), closed expressions for the perturbation contribution to the free energy and the chemical potentials can be presented. Finally, we offer suggestions for numerical implementations. PMID- 11031582 TI - Simple model for mixing at accelerated fluid interfaces with shear and compression AB - A simple model was recently described for predicting linear and nonlinear mixing at an unstable planar interface between two fluids of different density subjected to an arbitrary time-dependent variable acceleration history [J. D. Ramshaw, Phys. Rev. E 58, 5834 (1998)]. Here we generalize this model to include the Kelvin-Helmholtz (KH) instability resulting from a tangential velocity discontinuity delta u, as well as the effects of a uniform anisotropic compression or expansion of the mixing layer as a whole. The model consists of a second-order nonlinear ordinary differential equation of motion for the half width h of the mixing layer. This equation is derived by combining the wavelength renormalization hypothesis used in the earlier model with a suitable expression for the rate of change of the kinetic energy of the mixing layer. The resulting generalized model contains no additional free parameters, and reduces to the previous model in the absence of tangential velocities and compression. It also reduces in the linear regime to the correct linearized stability equation for an accelerated shear layer with compression [J. D. Ramshaw, Phys. Rev. E 61, 1486 (2000)]. For a pure incompressible KH instability in the nonlinear regime, the model predicts that h = eta magnitude of delta u t, where eta = [alpha(2 - theta)/square root of theta(1 - theta)]square root of rho 1 rho 2/(rho 1 + rho 2), and alpha and theta are parameters appearing in the nonlinear Rayleigh-Taylor and Richtmyer-Meshkov growth laws. For equal densities and the same parameter values previously used to match variable-acceleration experimental data, we find eta = 0.10, in close agreement with experimental data for free shear layers. PMID- 11031583 TI - Phonon self-energies and phase transitions in a prototype discotic liquid crystal AB - Specific Raman active vibrations in the discotic liquid crystal nematogen hexakis(6-hexyloxy)triphenylene are shown to be sensitive to either the isotropic<-->columnar or the columnar<-->solid phase transitions. Changes in frequency and/or intensity of vibrations with specific symmetries demonstrate that the method can be used not only to monitor the phase transitions themselves with a microscopic and noninvasive optical technique, but also to gain physical information on the origin of the molecular interactions that produce them. PMID- 11031584 TI - Quasicritical behavior of dielectric permittivity in the isotropic phase of smectogenic n-cyanobiphenyls AB - Results are presented of temperature and pressure studies of static dielectric permittivity (epsilon) and the nonlinear dielectric effect (NDE) in the isotropic phase of smectogenic n-cyanobiphenyls: 9CB, 10CB, and 12CB (4-cyano-4'-n alkylbiphenyl, n = 9, 10, and 12). For the mentioned properties, pretransitional effects can be well portrayed by applying the relations used for the isotropic phase of nematogens, where evidence of the quasicritical, fluidlike behavior with exponents alpha approximately 0.5 and gamma = 1 exists. This kind of behavior one can also observe on approaching the isotropic-smectic A transition. NDE studies in 10CB and 12CB made it possible to determine the pressure evolution of the discontinuity (delta T) of the I-SmA transition. It was found that pressure first decreases the discontinuity of the transition, and that next a gradual rise appears. This behavior is unlike the one observed for the isotropic-nematic transition, where only an increase of delta T with rising pressure was observed. PMID- 11031585 TI - Phase transitions from the isotropic liquid to liquid crystalline mesophases studied by linear and nonlinear static dielectric permittivity AB - Results of studies of static dielectric permittivity (epsilon) and nonlinear dielectric effect (NDE) in the isotropic phase of 4-n-4'-isothiocyanatobiphenyl (nBT) homologous series from n = 2 to n = 10 exhibiting the isotropic-smectic E (I-SmE) transition, are presented. They are compared with results of similar studies in 4-cyano-4-n-alkylbiphenyls (nCB) from n = 4 to n = 12. In this homologous series isotropic-nematic (I-N) and isotropic-smectic-A (I-Sm-A) transitions take place. Despite significant differences between N, Sm-A, and Sm-E phases the same pretransitional behavior of epsilon and NDE in the isotropic phase, described by critical exponents gamma = 1 and alpha = 0.5, was found. It has been shown that when the length of the alkyl chain of a compound increases the discontinuity of the transition drops in nBT and rises in nCB. The influence of pressure on the discontinuity is also discussed. PMID- 11031586 TI - Pseudo-Casimir effect in nematic liquid crystals in frustrating geometries AB - We study theoretically the fluctuation-induced structural force in nematic systems frustrated by external fields. We focus on the uniform director structure in the hybrid-aligned film characterized by opposing surface fields and in the Freedericksz cell where frustration arises from competing bulk and surface fields. We find that frustration gives rise to several interesting features of the interaction, including a crossover from attraction at small distances to repulsion at large distances. The fluctuation-induced interaction is enhanced substantially by frustration, the enhancement being progressively stronger on approaching the transition from uniform to distorted structure. At the structural transition the interaction diverges and we show that the pretransitional singularity is universal. PMID- 11031587 TI - Helix inversion in the chiral nematic and isotropic phases of a liquid crystal AB - Measurements of the chirality (2 pi/pitch) in the chiral nematic phase and of a structural constant proportional to the chirality in the isotropic liquid for a system in which a helix inversion line crosses the chiral nematic to isotropic phase transition line are reported. While the chirality shows a strong temperature dependence in the chiral nematic phase, it loses all temperature dependence in the isotropic phase. In addition, the chirality in the isotropic phase is proportional to the chirality in the chiral nematic phase at the phase transition, and may in fact be continuous across the transition. While molecular field and phenomenological theories can explain the strong temperature dependence in the chiral nematic phase, including the helix inversion, these theories predict a strong discontinuity in the chirality at the phase transition that is not supported by experiment. So while a theory that includes short range molecular correlations is called for to understand the behavior of the chirality across the phase transition, theoretical attempts to explain the chirality of a phase from a microscopic level must account for the strong role played by long range orientational order. PMID- 11031588 TI - Effect of photoisomerization of azobenzene dopants on the flexoelectric properties of short-pitch cholesteric liquid crystals AB - The flexoelectric properties of short-pitch cholesteric mixtures doped with three different azobenzenes, 4,4'-dihexyloxyazobenzene (4,4'-azo), 3,3'-dihexyloxy-2,2' dimethylazobenzene (3,3'-azol), and 3,3'-dihexanoyloxy-2,2'-dimethylazobenzene (3,3'-azo2), respectively, were studied upon illumination with uv light. Their effective flexoelectric coefficients were derived from the flexoelectro-optic response of the mixtures aligned in uniform lying helix texture. Considering the fact that the pitch of the mixtures became shorter upon uv illumination, an increase of their effective flexoelectric coefficients was found to take place due to the photoisomerization of the dyes. The largest change was found for the coefficient of the guest-host mixture containing 4,4'-azo dye, most probably due to the bent shape of the dye cis-isomer. This observation is in good agreement with our previous studies on the influence of the molecular shape on the liquid crystal flexoelectric properties and it suggests a possible way for enhancement of the amplitude of flexoelectro-optic response in cholesterics by using liquid crystal materials with pronounced molecular shape dissymmetry. PMID- 11031589 TI - Local field effects and the field-dependent dielectric response of polymer dispersed liquid crystals AB - An anisotropic version of the Maxwell Garnett approximation is applied for studying the dielectric properties of polymer dispersed liquid crystals containing bipolar liquid crystal droplets. This approach provides an explicit link between the droplet orientation distribution and the macroscopic response of the material. The electrostatic energy of the droplets is balanced with a strong anchoring elastic energy term for different initial orientation distributions. For aligned droplets we find a switching process whose sharpness depends on the initial orientation and a concentration dependent threshold field. For a planar distribution we find sharp transitions with a hysteresis loop whose width depends on the droplet concentration. For a random distribution the droplet reorientation is more gradual. The theory is also applied to the negligible elastic energy limit, recently observed at temperatures near the nematic-isotropic phase transition, where the droplets consist of bipolar nematic cores coated by isotropic liquid shells. This structural change within the droplets causes a considerable modification of the electro-optical properties. The Maxwell Garnett approach is used to calculate the dielectric response of this structure and reproduces all the main features of the experimental results. PMID- 11031590 TI - Dilution of nematic surface potentials: statics AB - The weak anchoring of a nematic liquid crystal is further illuminated by studying the consequences of finite-range surface torques. It is shown that the actual decay law of a diluted surface potential has little influence on either the equilibrium profile and the saturation field, provided that the range of the potential is small compared to the surface extrapolation length. This appears as a possible way to extend the conventional Rapini-Papoular model without altering its predictions away from the boundary. The net advantage of a dilution model is to better understand surface phenomena, which indeed pertain to a thin boundary layer. PMID- 11031591 TI - Crystalline transitions in free-standing films of 4-n-heptyloxybenzylidene-4-n heptylaniline AB - Free-standing films of 4-n-heptyloxybenzylidene-4-n-heptylaniline are known to exhibit multiple smectic-I surface layers in the presence of a smectic-A interior. The phase transitions leading to the crystallization of these films have been studied using electron diffraction. Our data are consistent with the scenario of the smectic-I surface layers first developing a crystal-B outermost layer before transforming entirely into the crystal-B phase, to be followed by the freezing of the smectic-A interior. The adjacent crystal-B and smectic-I layers show evidence of orientational epitaxy. PMID- 11031593 TI - Effect of chainlike aggregates on dynamical properties of magnetic liquids AB - The effect of chain-shaped aggregates on the dynamic properties of magnetic liquids is studied under the assumption that the chains can be modeled as straight and rigid. On the basis of microscopical analysis the macroscopical expression for average stress tensor is obtained. The effective viscosity and viscoelastic characteristics are estimated. PMID- 11031592 TI - Inversion in the change of the refractive index and memory effect near the nematic-isotropic phase transition in a lyotropic liquid crystal. AB - This work demonstrates the occurrence of dn/dT inversion from negative to positive near the nematic-isotropic phase transition in a lyotropic liquid crystal. It is suggested that this effect can be attributed to a sudden increase of the electronic polarizability due to a change in the micelle shape near this phase transition. Formation of a long lasting lenslike element within the sample when it is irradiated at moderately high laser powers is also reported. This permanent lens is erasable by increasing the temperature above the nematic isotropic transition temperature. PMID- 11031594 TI - Diffusion-limited aggregation: a revised mean-field approach AB - We propose a revision of the classic mean-field approach of diffusion-limited aggregation (DLA) model originally introduced by Witten and Sander [Phys. Rev. Lett. 47, 1400 (1981)]. The derived nonlinear mean-field equations providing lattice anisotropy are used to model diffusional growth on square lattice in linear and circular source geometries. The overall cluster shapes obtained from the mean-field calculations are found to satisfy the known scaling behavior experimentally observed for DLA simulations. PMID- 11031595 TI - Adsorption-desorption model and its application to vibrated granular materials. AB - We investigate both analytically and by numerical simulation the kinetics of a microscopic model of hard rods adsorbing on a linear substrate, a model that is relevant for compaction of granular materials. The computer simulations use an event-driven algorithm that is particularly efficient at very long times. For a small, but finite desorption rate, the system reaches an equilibrium state very slowly, and the long-time kinetics display three successive regimes: an algebraic one where the density varies as 1/t, a logarithmic one where the density varies as 1/ln(t), followed by a terminal exponential approach. The characteristic relaxation time of the final regime, though incorrectly predicted by mean field arguments, can be obtained with a systematic gap-distribution approach. The density fluctuations at equilibrium are also investigated, and the associated time-dependent correlation function exhibits a power law regime followed by a final exponential decay. Finally, we show that denser particle packings can be obtained by varying the desorption rate during the process. PMID- 11031596 TI - Tip-splitting instabilities in the channel Saffman-Taylor flow of constant viscosity elastic fluids AB - Boger fluids are used to study viscous fingering growth in viscoelastic fluids in channel Hele-Shaw flow. We have found that the viscous finger growing in the Boger fluid is unstable to tip splitting at high velocities, in a regime where a Newtonian viscous finger is stable. No fracturelike instabilities were observed. We show that the viscoelastic normal stress differences arising in shear and extensional flow reach very high values at shear and extensional rates comparable to those achieved at the tip of the finger at the onset of tip splitting, and the fluid becomes highly anisotropic. The viscoelastic stress could affect the dynamics of the finger and induce the tip-splitting instability. PMID- 11031597 TI - Colloidal particles in emulsions AB - We propose a statistical mechanical model for colloidal particles suspended in an emulsion of liquid droplets. The particles are modeled as hard spheres. The interaction between droplets is also hard, but the particles are able to penetrate the droplets. A swelling of droplets is taken into account to ensure material conservation of the droplet liquid. Hence the presence of the colloids generates droplet polydispersity. Using computer simulation and liquid state theory, we find that the relative polydispersity exhibits nonmonotonic behavior as a function of the particle packing fraction and can be traced back to hard sphere bulk density fluctuations. PMID- 11031598 TI - Internal structure of dense electrodeposits AB - We report experimental investigations of the structure of dense patterns obtained during electrochemical deposition of copper in thin cells. The deposit correlation function reveals the periodic structuration of the patterns but shows that the primary spacing is not steady during the growth and that moreover it is not simply related to the diffusion length. Another measurable quantity is the occupancy ratio of the fingers in the cell. Its variation as a function of the experimental parameters is interpreted from specific properties of electrochemical growth. The results are discussed with respect to the well-known behavior of cellular solidification fronts. PMID- 11031599 TI - A two-time-scale, two-temperature scenario for nonlinear rheology AB - We investigate a general scenario for "glassy" or "jammed" systems driven by an external, nonconservative force, analogous to a shear force in a fluid. In this scenario, the drive results in the suppression of the usual aging process, and the correlation and response functions become time translation invariant. The relaxation time and the response functions are then dependent on the intensity of the drive and on temperature. We investigate this dependence within the framework of a dynamical closure approximation that becomes exact for disordered, fully connected models. The relaxation time is shown to be a decreasing function of the drive ("shear thinning" effect). The correlation functions below the glass transition temperature (Tc) display a two-time-scale relaxation pattern, similar to that observed at equilibrium slightly above Tc. We also study the violation of the fluctuation-dissipation relationship in the driven system. This violation is very reminiscent of the one that takes place in a system aging below Tc at zero drive. It involves, in particular the appearance of a two-temperature regime, in the sense of an effective fluctuation-dissipation temperature [L. F. Cugliandolo, J. Kurchan, and L. Peliti, Phys. Rev. E 55, 3898 (1997)]. Although our results are, in principle, limited to the closure relations that hold for mean-field models, we argue that a number of the salient features are not inherent to the approximation scheme, and may be tested in experiments and simulations. PMID- 11031600 TI - Replica-exchange molecular dynamics simulation for supercooled liquids AB - We investigate to what extend the replica-exchange Monte Carlo method is able to equilibrate a simple liquid in its supercooled state. We find that this method does indeed allow us to generate accurately the canonical distribution function even at low temperatures and that its efficiency is about 10-100 times higher than the usual canonical molecular dynamics simulation. PMID- 11031601 TI - Slow viscous flows in micropolar fluids AB - A systematic calculation of micropolar fluid flows around a sphere and a cylinder is presented. The explicit velocity fields and the drag forces exerted by the fluid flow in both two and three dimensions are obtained. The solution of a steady micropolar fluid flow inside the cylinder is also obtained and is identical to the form observed in an experiment on granular vibrating beds. PMID- 11031602 TI - Independent ion migration in suspensions of strongly interacting charged colloidal spheres AB - We report on systematic measurements of the low-frequency conductivity sigma in aqueous suspensions of highly charged colloidal spheres. Sample preparation in a closed tubing system results in precisely controlled number densities of 10(16) m 3 < or = n < or = 10(19) m-3 (packing fractions of 10(-7) < or = phi < or = 10( 2)) and electrolyte concentrations of 10 < or = c < or = 10(-3) mol l-1. Due to long-range Coulomb repulsion, some of the systems show a pronounced fluid or crystalline order. Under deionized conditions we find sigma to depend linearly on the packing fraction with no detectable influence of the phase transitions. Further, at constant packing fraction sigma increases sublinearly with the increasing number of dissociable surface groups N. As a function of c the conductivity shows pronounced differences depending on the kind of electrolyte used. We propose a simple yet powerful model based on the independent migration of all species present and the additivity of the respective conductivity contributions. It takes account of small ion macro-ion interactions in terms of an effectively transported charge. The model successfully describes our qualitatively complex experimental observations. It further facilitates quantitative estimates of sigma over a wide range of particle and experimental parameters. PMID- 11031603 TI - Occurrence of coexisting dendrite morphologies: immiscible fluid displacement in an anisotropic radial Hele-Shaw cell under a high flow rate regime AB - Viscous fingering morphologies during the displacement of a high viscosity fluid by a low viscosity immiscible fluid in a radial fourfold anisotropic Hele-Shaw cell are examined. By using the kerosene-glycerin system for which the mu/T ratio (mu being the relative viscosity and T the interfacial tension between the fluids) is about ten times higher than that for the commonly used air-glycerin system, we have been able to access the hitherto unexplored Nca greater than approximately 1 regime (capillary number Nca = U mu/T, U being the advancing fingertip velocity). Within the anisotropy-dominated regime, and when flow rates are significantly high (capillary number well beyond Nca = 1), a new phase is seen to evolve wherein the dendrites grow simultaneously along the channels and along the directions making an angle of 45 degrees with the channels, both being kinetically driven. This new phase resembles the one observed in a miscible fluid system at all flow rates of the displacing fluid. [A. G. Banpurkar et al., Phys. Rev. E 59, 2188 (1999)]. PMID- 11031604 TI - Pattern selection induced by electroconvection in the electrodeposition of iron AB - The morphology of iron electrodeposit is shown to relate closely to the pH of the electrolyte solution. Macroscopically, depending on the strength of the interbranch convection, which is associated with the concentration of H3O+ in the electrolyte, the deposit morphology varies from treelike pattern to meshlike pattern and dense-branching morphology. Microscopically the deposit is ramified and dense-branching at lower concentration of H3O+, while it becomes relatively smooth and stringy at higher H3O+ concentration. The symmetry of the convective vortices on the two sides of the growing tip is observed to decide the growth behavior of the tip. We suggest that H3O+ influences the pattern formation and pattern selection in the electrodeposition of iron from FeSO4 solution by either initiating interbranch convection or changing the effective interfacial energy of the deposit and the electrolyte. PMID- 11031605 TI - Semiflexible polymer on an anisotropic Bethe lattice AB - The mean-square end-to-end distance of an N-step polymer on a Bethe lattice is calculated. We consider semiflexible polymers placed on isotropic and anisotropic lattices. The distance on the Cayley tree is defined by embedding the tree on a sufficiently high-dimensional Euclidean space, considering that every bend of the polymer defines a direction orthogonal to all the previous ones. In the isotropic case, the result obtained for the mean-square end-to-end distance turns out to be identical to the one obtained for ideal chains without immediate returns on an hypercubic lattice with the same coordination number of the Bethe lattice. For the general case, we obtain asymptotic behavior in both the semiflexible and almost rigid limits. PMID- 11031606 TI - Wave number of maximal growth in viscous magnetic fluids of arbitrary depth AB - An analytical method within the frame of linear stability theory is presented for the normal field instability in magnetic fluids. It allows us to calculate the maximal growth rate and the corresponding wave number for any combination of thickness and viscosity of the fluid. Applying this method to magnetic fluids of finite depth, these results are quantitatively compared to the wave number of the transient pattern observed experimentally after a jumplike increase of the field. The wave number grows linearly with increasing induction where the theoretical and the experimental data agree well. Thereby, a long-standing controversy about the behavior of the wave number above the critical magnetic field is tackled. PMID- 11031607 TI - Internal dynamics and elasticity of confined entropic gels AB - The internal dynamics and elasticity of colloidal gels formed via depletion in confined nearly hard-sphere mixtures are studied via particle-tracking measurements of the self part of the van Hove correlation function, which is found to decay exponentially in space and exhibit a scaling in terms of its first moment. The second moment, or mean-square particle displacement, exhibits a stretched-exponential decay to a plateau and provides a measure of the weak elastic modulus of the gel. A Fourier analysis of the real-space probability distribution yields the intermediate scattering function, which at fixed wave vector exhibits a stretched exponential decay to a nonzero plateau, reminiscent of dynamic behavior observed in a broad class of disordered systems. PMID- 11031608 TI - Tightness of random knotting. AB - Long polymers in solution frequently adopt knotted configurations. To understand the physical properties of knotted polymers, it is important to find out whether the knots formed at thermodynamic equilibrium are spread over the whole polymer chain or rather are localized as tight knots. We present here a method to analyze the knottedness of short linear portions of simulated random chains. Using this method, we observe that knot-determining domains are usually very tight, so that, for example, the preferred size of the trefoil-determining portions of knotted polymer chains corresponds to just seven freely jointed segments. PMID- 11031609 TI - Elasticity of Poissonian fiber networks AB - An effective-medium model is introduced for the elasticity of two-dimensional random fiber networks. These networks are commonly used as basic models of heterogeneous fibrous structures such as paper. Using the exact Poissonian statistics to describe the microscopic geometry of the network, the tensile modulus can be expressed by a single-parameter function. This parameter depends on the network density and fiber dimensions, which relate the macroscopic modulus to the relative importance of axial and bending deformations of the fibers. The model agrees well with simulation results and experimental findings. We also discuss the possible generalizations of the model. PMID- 11031610 TI - Mach cone shocks in a two-dimensional Yukawa solid using a complex plasma AB - Mach cones were studied experimentally in a two-dimensional Yukawa solid consisting of charged micrometer particles suspended as a layer in a plasma. These cones were V-shaped shocks produced spontaneously by a supersonic particle moving below the main two-dimensional particle layer. The cones had a double structure. The first cone was compressional and particles moved forward, and it was followed by a second cone, which was rarefactional, where particles moved backward. Over the limited range of speed V attained by the supersonic particles in this experiment, the angle mu of the cone was found to obey the Mach cone rule sin mu = c/V, where c is the medium's sound speed. The cones caused only elastic deformations in the crystal lattice, except in a narrow track behind the cone's vertex. The wings of the cones can be analyzed as linear shocks in two dimensions. Using spatially resolved measurements of the particle number density and velocity and applying the Hugoniot relations for shocks in two dimensions, we found that the pressure inside the first Mach cone was greater than in the undisturbed medium by a factor of 1.3-1.6. The cone angle was also used to measure the charge in this experiment. PMID- 11031611 TI - Velocity distribution in granular gases of viscoelastic particles AB - The velocity distribution in a homogeneously cooling granular gas has been studied in the viscoelastic regime, when the restitution coefficient of colliding particles depends on the impact velocity. We show that for viscoelastic particles a simple scaling hypothesis is violated, i.e., that the time dependence of the velocity distribution does not scale with the mean square velocity as in the case of particles interacting via a constant restitution coefficient. The deviation from the Maxwellian distribution does not depend on time monotonically. For the case of small dissipation we detected two regimes of evolution of the velocity distribution function: Starting from the initial Maxwellian distribution, the deviation first increases with time on a collision time scale saturating at some maximal value; then it decays to zero on a much larger time scale which corresponds to the temperature relaxation. For larger values of the dissipation parameter there appears an additional intermediate relaxation regime. Analytical calculations for small dissipation agree well with the results of a numerical analysis. PMID- 11031612 TI - Wavelength scaling of spiral patterns formed by granular media underneath a rotating fluid AB - A spiral pattern formed by granular media underneath a rotating fluid is discussed. Results from a cellular-automaton model are compared to experimental data, and are found to reproduce experimentally observed scalings. A theoretical argument predicting these scalings on the basis of the existence of a critical threshold condition is advanced. It is suggested that the pattern is probably not associated with a hitherto unknown flow instability, as has been speculated previously. It appears that the pattern constitutes some rotating analog to sand ripples in nonrotating systems. PMID- 11031613 TI - Symmetric heaping in grains: a phenomenological model AB - Heap formation of granular materials in a vertical vibrating bed is studied by a simple model using the profile of the heap as the dynamic variable. Vibration increases the local height, but is counterbalanced by the nonlinear coupling, which tends to suppress the growth of the height. The steady state heap can be solved in closed form in terms of Jacobian elliptic functions. Phenomena such as heap formation and downward and upward heaps can be reproduced. Our results agree with the experimentally observed change of downward to upward steady heaps as the vibration strength is increased. Predictions from the model compare favorably with experimental results on heap profiles and heaping angles. PMID- 11031614 TI - Patterns in thin vibrated granular layers: interfaces, hexagons, and superoscillons AB - A theoretical and experimental study of patterns in vibrated granular layers is presented. An order parameter model based on the parametric Ginzburg-Landau equation is used to describe strongly nonlinear excitations including hexagons, interfaces between flat antiphase domains, and new localized objects, superoscillons. The experiments confirm the existence of superoscillons and bound states of superoscillons and interfaces. On the basis of the order parameter model we predict analytically and confirm experimentally that additional subharmonic driving results in the controlled motion of interfaces. PMID- 11031615 TI - Enhancement of epidemic spread by noise and stochastic resonance in spatial network models with viral dynamics. AB - We extend a previous dynamical viral network model to include stochastic effects. The dynamical equations for the viral and immune effector densities within a host population of size n are bilinear, and the noise is white, additive, and Gaussian. The individuals are connected with an n x n transmission matrix, with terms which decay exponentially with distance. In a single individual, for the range of noise parameters considered, it is found that increasing the amplitude of the noise tends to decrease the maximum mean virion level, and slightly accelerate its attainment. Two different spatial dynamical models are employed to ascertain the effects of environmental stochasticity on viral spread. In the first model transmission is unrestricted and there is no threshold within individuals. This model has the advantage that it can be analyzed using a Fokker Planck approach. The noise is found both to synchronize and uniformize the trajectories of the viral levels across the population of infected individuals, and thus to promote the epidemic spread of the virus. Quantitative measures of the speed of spread and overall amplitude of the epidemic are obtained as functions of the noise and virulence parameters. The mean amplitude increases steadily without threshold effects for a fixed value of the virulence as the noise amplitude sigma is increased, and there is no evidence of a stochastic resonance. However, the speed of transmission, both with respect to its mean and variance, undergoes rapid increases as sigma changes by relatively small amounts. In the second, more realistic, model, there is a threshold for infection and an upper limit to the transmission rate. There may be no spread of infection at all in the absence of noise. With increasing noise level and a low threshold, the mean maximum virion level grows quickly and shows a broad-based stochastic resonance effect. When the threshold within individuals is increased, the mean population virion level increases only slowly as sigma increases, until a critical value is reached at which the mean infection level suddenly increases. Similar results are obtained when the parameters of the model are also randomized across the population. We conclude with a discussion and a description of a diffusion approximation for a model in which stochasticity arises through random contacts rather than fluctuation in ambient virion levels. PMID- 11031616 TI - Statistical analysis of genealogical trees for polygamic species. AB - Repetitions within a given genealogical tree provide some information about the degree of consanguinity of a population. They can be analyzed with techniques usually employed in statistical physics when dealing with fixed point transformations. In particular, we show that the tree features strongly depend on the fractions of males and females in the population, and also on the offspring probability distribution. We check different possibilities, some of them relevant to human groups, and compare them with simulations. PMID- 11031617 TI - Species independence of mutual information in coding and noncoding DNA. AB - We explore if there exist universal statistical patterns that are different in coding and noncoding DNA and can be found in all living organisms, regardless of their phylogenetic origin. We find that (i) the mutual information function [symbol: see text] has a significantly different functional form in coding and noncoding DNA. We further find that (ii) the probability distributions of the average mutual information [symbol: see text] are significantly different in coding and noncoding DNA, while (iii) they are almost the same for organisms of all taxonomic classes. Surprisingly, we find that [symbol: see text] is capable of predicting coding regions as accurately as organism-specific coding measures. PMID- 11031618 TI - Exact equations and scaling relations for f0 avalanche in the Bak-Sneppen evolution model. AB - An infinite hierarchy of exact equations is derived for the newly observed f0 avalanche in the Bak-Sneppen model. By solving the first-order exact equation, we find that the critical exponent gamma, governing the divergence of the average avalanche size, is exactly 1 (for all dimensions), which has been confirmed by extensive simulations. Solution of the gap equation yields another universal result rho = 1 (rho is the exponent of relaxation to attractor). Scaling relations are established among the critical exponents (gamma, tau, D, sigma, and nu) for the f0 avalanche. PMID- 11031619 TI - Critical swelling in single phospholipid bilayers. AB - We approach the controversial anomalous swelling problem in membrane systems using small angle neutron scattering to measure relative changes in the bilayer thickness of unilamellar vesicles of dimyristoylphosphatidylcholine lipid bilayers in the vicinity of the main transition. These measurements conclusively demonstrate that at least half of the anomalous swelling previously observed in multilamellar vesicles of this system can be accounted for by the critical thickening of the bilayer itself, in contrast to conclusions drawn from several recent studies. PMID- 11031620 TI - Base opening in RNA and DNA duplexes: implication for RNA stability. AB - The energetics of a low-energy single base opening in several RNA duplex crystal structures has been calculated and compared to DNA duplexes. Base opening in RNA appears to have an overall preference towards the major groove, similar to results previously reported for B-DNA. Movement of each of the adenine, uracil, and cytosine bases into the minor groove is blocked by a high-energy barrier due to severe close contact with neighboring bases. Guanine bases are able to open towards both grooves because of the unique orientation of the base that avoids steric clash along the opening pathway. RNA bases are found to have a substantially smaller major groove opening extent than that of their B-DNA counterparts. A comparison with base opening behavior of A-DNA duplexes suggests that this difference results from helix constraint associated with A-form backbone conformation. The reduced opening extent correlates with the RNA duplex stability and is consistent with observed slower imino proton exchange rates in RNA duplexes. PMID- 11031621 TI - Microrheometry underestimates the values of the viscoelastic moduli in measurements on F-actin solutions compared to macrorheometry. AB - We present a systematic comparison of microrheological and macrorheological measurements of the viscoelastic storage and loss moduli, G'(f) and G"(f), respectively, of solutions of the semiflexible biopolymer F-actin. Using magnetic tweezers microrheometry and rotating disk macrorheometry, we show that microscopic values for G'(f) and G"(f) are significantly smaller than macroscopic results over the frequency range f = 0.004-4 Hz, whereas the qualitative shape of the spectra is similar. These findings confirm recent theoretical predictions [A. C. Maggs, Phys. Rev. E 57, 2091 (1998)]. The discrepancy affects not only absolute values of G'(f) and G"(f): although microscopic and macroscopic plateau regime are found in the same frequency range, the two methods yield different values for the entanglement time which determines the high-frequency end of the plateau. By investigating F-actin solutions of different mean filament lengths, we show that microscopic and macroscopic G'(f) and G"(f) converge, if the probe particle used in microrheometry becomes large compared to the length of actin filaments. PMID- 11031622 TI - Noise-induced memory in extended excitable systems. AB - We describe a form of memory exhibited by extended excitable systems driven by stochastic fluctuations. Under such conditions, the system self-organizes into a state characterized by power-law correlations, thus retaining long-term memory of previous states. The exponents are robust and model independent. We discuss implications of these results for the functioning of cortical neurons as well as for networks of neurons. PMID- 11031623 TI - Mean field theory for asymmetric neural networks. AB - The computation of mean firing rates and correlations is intractable for large neural networks. For symmetric networks one can derive mean field approximations using the Taylor series expansion of the free energy as proposed by Plefka. In asymmetric networks, the concept of free energy is absent. Therefore, it is not immediately obvious how to extend this method to asymmetric networks. In this paper we extend Plefka's approach to asymmetric networks and in fact to arbitrary probability distributions. The method is based on an information geometric argument. The method is illustrated for asymmetric neural networks with sequential dynamics. We compare our approximate analytical results with Monte Carlo simulations for a network of 100 neurons. It is shown that the quality of the approximation for asymmetric networks is as good as for symmetric networks. PMID- 11031624 TI - Time evolution of the Partridge-Barton model. AB - The time evolution of the Partridge-Barton model in the presence of the pleiotropic constraint and deleterious somatic mutations is exactly solved for arbitrary fecundity in the context of a matricial formalism. Analytical expressions for the time dependence of the mean survival probabilities are derived. Using the fact that the asymptotic behavior for large time t is controlled by the largest matrix eigenvalue, we obtain the steady state values for the mean survival probabilities and the Malthusian growth exponent. The mean age of the population exhibits a t-1 power law decayment. Some Monte Carlo simulations were also performed and they corroborated our theoretical results. PMID- 11031625 TI - Clarification of the ripple phase of lecithin bilayers using fully hydrated, aligned samples. AB - Aligned samples of lipid bilayers have been fully hydrated from water vapor in a different type of x-ray chamber. Our use of aligned samples resolves issues concerning the ripple phase that were ambiguous from previous powder studies. In particular, our x-ray diffraction data conclusively demonstrate that, on cooling from the L alpha to the P beta' phase, both chiral and racemic samples of dipalmitoyl phosphatidylcholine (DPPC) exhibit phase coexistence of long and short ripples with a ripple wavelength ratio lambda L/lambda S approximately 1.8. Moreover, the long ripple always forms an orthorhombic unit cell (gamma L = 90 degrees), strongly supporting the possibility that these ripples are symmetric. In contrast, gamma S for short ripples was consistently different from 90 degrees, implying asymmetric ripples. We continue to find no evidence that chirality affects the structure of rippled bilayers. The relative thermodynamic stability of the two types of ripples was investigated and a qualitative free energy diagram is given in which the long ripple phase is metastable. Finally, we suggest a kinetic mechanism, involving loss of water, that promotes formation of the metastable long ripple phase for special thermal protocols. PMID- 11031626 TI - Epidemics and percolation in small-world networks. AB - We study some simple models of disease transmission on small-world networks, in which either the probability of infection by a disease or the probability of its transmission is varied, or both. The resulting models display epidemic behavior when the infection or transmission probability rises above the threshold for site or bond percolation on the network, and we give exact solutions for the position of this threshold in a variety of cases. We confirm our analytic results by numerical simulation. PMID- 11031627 TI - Enhanced coding in a cochlear-implant model using additive noise: aperiodic stochastic resonance with tuning. AB - Analog electrical stimulation of the cochlear nerve (the nerve of hearing) by a cochlear implant is an effective method of providing functional hearing to profoundly deaf people. Recent physiological and computational experiments have shown that analog cochlear implants are unlikely to convey certain speech cues by the temporal pattern of evoked nerve discharges. However, these experiments have also shown that the optimal addition of noise to cochlear implant signals can enhance the temporal representation of speech cues [R. P. Morse and E. F. Evans, Nature Medicine 2, 928 (1996)]. We present a simple model to explain this enhancement of temporal representation. Our model derives from a rate equation for the mean threshold-crossing rate of an infinite set of parallel discriminators (level-crossing detectors); a system that well describes the time coding of information by a set of nerve fibers. Our results show that the optimal transfer of information occurs when the threshold level of each discriminator is equal to the root-mean-square noise level. The optimal transfer of information by a cochlear implant is therefore expected to occur when the internal root-mean square noise level of each stimulated fiber is approximately equal to the nerve threshold. When interpreted within the framework of aperiodic stochastic resonance, our results indicate therefore that for an infinite array of discriminators, a tuning of the noise is still necessary for optimal performance. This is in contrast to previous results [Collins, Chow, and Imhoff, Nature 376, 236 (1995); Chialvo, Longtin, and Muller-Gerking, Phys. Rev. E 55, 1798 (1997)] on arrays of FitzHugh-Nagumo neurons. PMID- 11031628 TI - Soft-x-ray laser scheme in a plasma created by optical-field-induced ionization of nitrogen AB - An x-ray laser scheme based on the recombination of a fully stripped nitrogen plasma is presented. Plasma is assumed to be created by the optical-field ionization of a nitrogen gas jet of 10(19) cm-3 atomic density by an ultrashort (60 fs), high-intensity (3 x 10(19) W/cm2) Ti:sapphire laser. Results of two dimensional particle-in-cell simulations, modeling laser-plasma interaction, parametric heating, and ponderomotive effects are presented. Hydrodynamic and kinetics calculations are performed and predict important local gain for H-like nitrogen transitions at 25 and 134 A, following fast collisional recombination for specific plasma conditions. PMID- 11031629 TI - Temperature and impurity transport studies of heated tokamak plasmas by means of a collisional-radiative model of x-ray emission from Mo30+ to Mo39+ AB - This work presents and interprets, by means of detailed atomic calculations, observations of L-shell (n = 3-->n = 2) transitions in highly ionized molybdenum, the main intrinsic heavy impurity in the Frascati tokamak upgrade plasmas. These hot plasmas were obtained by additional electron cyclotron resonance heating (ECRH), at the frequency of 140 Ghz, during the current ramp-up phase of the discharge. Injecting 400 kW on axis and 800 kW slightly off axis, the peak central electron temperature reached 8.0 and 7.0 keV, respectively, for a time much longer than the ionization equilibrium time of the molybdenum ions. X-ray emissions from rarely observed high charge states, Mo30+ to Mo39+, have been studied with moderate spectral resolution (lambda/delta lambda approximately 150) and a time resolution of 5 ms. A sophisticated collisional-radiative model for the study of molybdenum ions in plasmas with electron temperature in the range 4 20 keV is presented. The sensitivity of the x-ray emission to the temperature and to impurity transport processes is discussed. This model has been then used to investigate two different plasma scenarios. In the first regime the ECRH heating occurs on axis during the current ramp up phase, when the magnetic shear is evolving from negative to zero up to the half radius. The spectrum is well reproduced with the molybdenum ions in coronal equilibrium and with a central impurity peaking. In the second regime, at the beginning of the current flat top when magnetic shear is monotonic and sawtoothing activity is appearing, the lowest charge states (Mo33+ to Mo30+), populated off axis, are affected by anomalous transport and the total molybdenum profile is found to be flat up to the half radius. We conclude with the presentation of "synthetic spectra" computed for even higher temperature plasmas that are expected in future experiments with higher ECRH power input. PMID- 11031630 TI - Nonlinear density wave theory for the spiral structure of galaxies. AB - The theory of nonlinear waves for plasmas has been applied to the analysis of the density wave theory of galaxies which are many-body systems of gravity. A nonlinear Schrodinger equation has been derived by applying the reductive perturbation method on the fluid equations that describe the behavior of infinitesimally thin disk galaxies. Their spiral arms are characterized by a soliton and explained as a pattern of a propagating nonlinear density wave. PMID- 11031631 TI - Electron density separations in nonideal plasmas: structure of Thomas-Fermi-like bound states and the Mott transition AB - Starting from a general free energy functional, the chemical picture for nonideal plasmas arises from a certain separation of the electron density. Defining a bound state and its electrostatically screening plasma environment as a subsystem, a simple theory for plasma-correspondent bound state structure can be formulated. This provides the possibility to derive adjustable parameters for the interactions in the chemical picture. The Mott transition corresponds to the violation of the normalization condition due to plasma influences. PMID- 11031632 TI - Explanations for the observed increase in fast electron penetration in laser shock compressed materials AB - We analyze recent experimental results on the increase of fast electron penetration in shock compressed plastic [Phys. Rev. Lett. 81, 1003 (1998)]. It is explained by a combination of stopping power and electric field effects, which appear to be important even at laser intensities as low as 10(16) W cm-2. An important conclusion is that fast electron induced heating must be taken into account, changing the properties of the material in which the fast electrons propagate. In insulators this leads to a rapid insulator to conductor phase transition. PMID- 11031633 TI - Hydrogen dissociation in a H2-N2 pulsed dc glow discharge AB - The absolute concentration of hydrogen atoms is measured in the positve column of a pulsed dc discharge by two-photon absorption laser-induced fluorescence in H2 and H2-N2 gas mixtures at constant pressure and current. The discharge pulse duration is varied from 10 microseconds to 1 msec. For shorter pulse durations of 10 and 100 microseconds, the H-atom signal decreased monotonically with H2 concentration; for longer pulse durations of > or = 500 microseconds, the fractional dissociation of the H2 is enhanced with increasing N2 concentration. The change in H-atom production from direct electron impact dissociation of H2 at short times compared to that from multiquantum vibrational energy transfer induced dissociation of H2 at long times is determined from temporally resolved H atom concentration measurements. PMID- 11031634 TI - Smith-Purcell radiation emission in aligned nanoparticles AB - Smith-Purcell (SP) radiation produced by interaction of fast electron beams running parallel to strings of nanoparticles is investigated. Results for the radiation emission probability and electron energy loss spectra using finite and infinite strings of Al and silica spheres are presented. Both of these quantities are obtained by solving Maxwell's equations exactly using a multipole expansion approach. The response of the spheres is described in terms of their local frequency-dependent dielectric functions. In silica, the emission probability is seen to coincide with the energy loss probability within the gap region, where the solid cannot absorb any energy. Large emission rates are predicted for Al, suggesting its possible application in tunable soft uv light generation. The dependence of the emission on the size of the spheres, the string period, and the electron energy is discussed in detail. Finite size effects are also studied for strings of 1-15 Al spheres. PMID- 11031635 TI - Production of halo particles by excitation of collective modes in high-intensity charged particle beams AB - This paper examines analytically and numerically the effects of self-consistent collective oscillations excited in a high-intensity charged particle beam on the motion of a test particle in the beam core. Even under ideal conditions, assuming a constant transverse focusing force (smooth focusing approximation), and perturbations about a uniform-density, constant radius beam, it is found that collective mode excitations, in combination with the applied focusing force and the equilibrium test fields, can eject particles from the beam core to large radii. Test particle orbits are calculated for collective oscillations with n = 1 and 2 radial mode structure, and an estimate is obtained for the range of initial conditions for which particles will be expelled from the beam interior. Resonances for meridional particles are found to be unimportant, while a class of particles with nonzero angular momentum are found to participate in resonant behavior. Once expelled from the beam, numerical solutions of the orbit equations indicate that Kolmogorov-Arnold-Moser curves, phase space spanning integrals of motion, confine particles within 1.5 times the beam radius for moderately low mode amplitudes, but are successively destabilized for higher amplitudes. PMID- 11031636 TI - Theory of Cerenkov and transition radiation from layered structures AB - A scattering theoretical approach is used to describe the generation of both transition and Cerenkov radiation due to the passage of fast electrons through layered, in general absorptive, dielectrics. It leads to a considerable improvement over the coherent summation method usually employed. Reflection of the produced radiation from the layer interfaces is now properly taken into account and absorption of radiation, if present, is automatically taken care of. The usual restriction that the energy of the produced photons must be small relative to the initial electron energy is lifted. In contrast to existing theories the production of Cerenkov radiation, if it takes place, is included as well. Our expressions for radiation production feature the eigenmodes of the Helmholtz equation for the dielectric and we discuss how the latter can be obtained by means of a transfer matrix formalism. A numerical evaluation shows that under the appropriate conditions our results are in agreement with those from the coherent summation formalism. Finally, we present numerical results that give an impression of the relative yields for transition and Cerenkov radiation. PMID- 11031637 TI - Ultrashort free-electron laser pulse AB - Three-dimensional characteristics of short free-electron laser pulses are analyzed. When the optical pulse length is short, the growth rate and optical guiding will vary among the Fourier components comprising the pulse. Matched beam solutions of the wave equation, including diffraction and nonparaxial effects, are discussed. In certain limits a front to back asymmetry develops along the pulse as well as a frequency spread across it. In these limits the asymmetry and the frequency spread are relatively small unless the number of optical cycles in the pulse approaches unity. PMID- 11031638 TI - Manifestation of intrinsic defects in optical properties of self-organized opal photonic crystals AB - Self-organized synthetic opals possessing a face centered cubic (fcc) lattice are promising for fabrication of a three-dimensional photonic crystal with a full photonic band gap in the visible. The fundamental limiting factor of this method is the large concentration of lattice defects and, especially, planar stacking faults, which are intrinsic to self-assembling growth of colloidal crystal. We have studied the influence of various types of defects on photonic band structure of synthetic opals by means of optical transmission, reflection and diffraction along different crystallographic directions. We found that in carefully chosen samples the stacking probability alpha can be as high as 0.8-0.9 revealing the strong preference of fcc packing sequence over the hexagonal close-packed (hcp). It is shown that scattering on plane stacking faults located perpendicular to the direction of growth results in a strong anisotropy of diffraction pattern as well as in appearance of a pronounced doublet structure in transmission and reflection spectra taken from the directions other than the direction of growth. This doublet is a direct manifestation of the coexistence of two crystallographic phases--pure fcc and strongly faulted. As a result the inhomogeneously broadened stop-bands overlap over a considerable amount of phase space. The latter, however, does not mean the depletion of the photonic density of states since large disordering results in filling of the partial gaps with both localized and extended states. PMID- 11031639 TI - Soliton formation from a pulse passing the zero-dispersion point in a nonlinear Schrodinger equation AB - We consider in detail the self-trapping of a soliton from a wave pulse that passes from a defocusing region into a focusing one in a spatially inhomogeneous nonlinear waveguide, described by a nonlinear Schrodinger equation in which the dispersion coefficient changes its sign from normal to anomalous. The model has direct applications to dispersion-decreasing nonlinear optical fibers, and to natural waveguides for internal waves in the ocean. It is found that, depending on the (conserved) energy and (nonconserved) "mass" of the initial pulse, four qualitatively different outcomes of the pulse transformation are possible: decay into radiation; self-trapping into a single soliton; formation of a breather; and formation of a pair of counterpropagating solitons. A corresponding chart is drawn on a parametric plane, which demonstrates some unexpected features. In particular, it is found that any kind of soliton(s) (including the breather and counterpropagating pair) eventually decays into pure radiation with an increase of energy, the initial "mass" being kept constant. It is also noteworthy that a virtually direct transition from a single soliton into a pair of symmetric counterpropagating ones seems possible. An explanation for these features is proposed. In two cases when analytical approximations apply, viz., a simple perturbation theory for broad initial pulses and the variational approximation for narrow ones, comparison with direct simulations shows reasonable agreement. PMID- 11031640 TI - Density of states for a dielectric superlattice. II. TM polarization AB - We present an analysis of the band structure, the equifrequency surfaces, and the density of states (DOS) for the transverse magnetic (TM) polarization mode of the dielectric superlattice, modeled by means of Dirac-delta functions. This complements a recent article [Phys. Rev E 59, 3624 (1999)] that analyzes the case of transverse electric (TE) polarization. Unfortunately, for this simple model, there is no manifestation of the Brewster effect in the band structure for the TM modes. For large values of the frequency or the grating strength, the equifrequency surfaces essentially degenerate into a set of concentric, hollow, and narrow cylinders centered on the superlattice axis. The DOS is enhanced relative to free space for any frequency and it exhibits discontinuities in the slope at the band edges. These results are relevant to the spontaneous emission by an atom or to dipole radiation in one-dimensional periodic structures. The differences between TE and TM modes are discussed. We take the opportunity to correct an error in the DOS calculation for TE polarization in the article referred above. PMID- 11031641 TI - Compression of femtosecond light pulses by one-dimensional photonic crystals with two-component relaxing nonlinearity AB - The compression of ultrashort light pulses in a one-dimensional nonlinear photonic crystal is investigated for light frequencies lying outside the forbidden gap of a periodic structure taking into account the delayed nonlinear response of a medium. It is shown that the relatively slow-responding defocusing nonlinearity can suppress the distortion of Bragg solitons caused by the relaxation of the fast self-focusing component and improve the compression efficiency. Stable optical quasisoliton propagation of light pulses and their compression are numerically confirmed for both band gap composite materials and photonic crystals with alternating layers of different types of nonlinearity. PMID- 11031642 TI - Effect of frequency detunings and finite relaxation rates on laser localized structures AB - We study, analytically and numerically, the effect of frequency detunings and relaxation processes in laser media on stability and bifurcations of dissipative optical localized structures (DOLS's) in a transversely one-dimensional laser with a saturable absorber. The approximate envelope equation, with an intensity dependent effective coefficient of the diffusion, is derived. Andronov-Hopf bifurcations resulting from frequency detuning and leading to oscillatory DOLS's are analyzed numerically. A numerical and analytical study of bifurcations of transversely motionless DOLS's in a laser with finite relaxation rates of amplifying and absorbing media is performed. New types of DOLS's are found, including those moving with a large transverse velocity and those moving with a periodically oscillating transverse velocity. Hysteresis between different types of DOLS's is demonstrated. PMID- 11031643 TI - Pulse-driven switching in one-dimensional nonlinear photonic band gap materials: a numerical study AB - We numerically examine the time-dependent properties of nonlinear bistable multilayer structures for constant wave illumination. We find that our system exhibits both steady-state and self-pulsing solutions. In the steady-state regime, we examine the dynamics of driving the system between different transmission states by injecting pulses, and we find optimal pulse parameters. We repeat this work for the case of a linear periodic system with a nonlinear impurity layer. PMID- 11031644 TI - Stability analysis for extended models of gap solitary waves AB - A numerical linear stability analysis has been carried out for stationary spatially localized solutions of several systems of coupled nonlinear partial differential equations (PDE's) with two and more complex variables. These coupled PDE's have recently been discussed in the literature, mostly in the context of physical systems with a frequency gap in the dispersion relation of their linear excitations, and they are extensions of the Mills-Trullinger gap soliton model. Translational and oscillatory instabilities are identified, and their associated growth rates are computed as functions of certain parameters characterizing the solitary waves. PMID- 11031645 TI - Perturbative study of classical Ablowitz-Ladik type soliton dynamics in relation to energy transport in alpha-helical proteins. AB - Classical Ablowitz-Ladik type soliton dynamics from three closely related classical nonlinear equations is studied using a perturbative method. Model nonintegrable equations are derived by assuming nearest neighbor hopping of an exciton(vibron) in the presence of a full exciton(vibron)-phonon interaction in soft molecular chains in general and spines of alpha-helices in particular. In all cases, both trapped and moving solitons are found implying activation energy barrier for propagating solitons. Analysis further shows that staggered and nearly staggered trapped solitons will have a negative effective mass. In some models the exciton(vibron)-phonon coupling affects the hopping. For these models, when the conservation of probability is taken into account, only propagating solitons with a broad profile are found to be acceptable solutions. Of course, for the soliton to be a physically meaningful entity, total nonlinear coupling strength should exceed a critical value. On the basis of the result, a plausible modification in the mechanism for biological energy transport involving conformational change in alpha-helix is proposed. Future directions of the work are also mentioned. PMID- 11031646 TI - Nonlinear Schrodinger flow in a periodic potential AB - We report a study of solutions of the defocusing nonlinear Schrodinger equation in a spatially periodic potential. The ground-state solution and the steady flows of the system are studied analytically. Above a critical current, a steady state no longer exists and time-dependent solutions are generated, which are numerically simulated and described. PMID- 11031647 TI - Growth and decay of discrete nonlinear Schrodinger breathers interacting with internal modes or standing-wave phonons AB - We investigate the long-time evolution of weakly perturbed single-site breathers (localized stationary states) in the discrete nonlinear Schrodinger equation. The perturbations we consider correspond to time-periodic solutions of the linearized equations around the breather, and can be either (i) spatially localized or (ii) spatially extended. For case (i), which corresponds to the excitation of an internal mode of the breather, we find that the nonlinear interaction between the breather and its internal mode always leads to a slow growth of the breather amplitude and frequency. In case (ii), corresponding to interaction between the breather and a standing-wave phonon, the breather will grow provided that the wave vector of the phonon is such that the generation of radiating higher harmonics at the breather is possible. In other cases, breather decay is observed. This condition yields a limit value for the breather frequency above which no further growth is possible. We also discuss another mechanism for breather growth and destruction which becomes important when the amplitude of the perturbation is non-negligible, and which originates from the oscillatory instabilities of the nonlinear standing-wave phonons. PMID- 11031648 TI - Kink-breather solution in the weakly discrete Frenkel-Kontorova model AB - The discrete Frenkel-Kontorova model, having the sine-Gordon equation as the continuous analog, was investigated numerically at a small degree of discreteness. Interaction between a kink and a breather in a discrete system was compared with the exact three-soliton solution to the continuous sine-Gordon equation. Nontrivial effects of discreteness were found numerically. One is that a kink and a breather in the discrete system are mutually attractive quasiparticles, so they can be regarded as a three-soliton oscillatory system. The other is the energy exchange between a kink and a breather when their collision takes place in a vicinity of a separatrix solution to the continuous sine-Gordon equation. We have estimated numerically the kink-breather binding energy EB and the maximum possible exchange energy EE for different breather frequencies omega. The results suggest that there is a threshold breather frequency for the "spontaneous" breaking up of the three-soliton oscillatory system into a kink and a breather moving in opposite directions. PMID- 11031649 TI - Instability of multiple pulses in coupled nonlinear Schrodinger equations AB - An analytical technique for constructing multiple pulses is presented in this article; with it we uncover a homoclinic bifurcation through which multihumped solitary waves can be generated in systems of coupled nonlinear Schrodinger equations. A method is then developed to determine the (in)stability of multiple pulses produced by this mechanism. The analysis is applied to two models that describe optical phenomena in dispersive quadratic and Kerr media, respectively. It sheds considerable light upon the characteristics that predispose multiple pulses arising in this class of systems to be unstable. PMID- 11031650 TI - Multisoliton complexes on a background AB - We obtain solutions of M coupled nonlinear Schrodinger equations that describe multisoliton complexes (MCs) on a background. We present explicit multiparameter families of solutions and numerical simulations, demonstrating specific features of MCs and their collisions. It is shown, in particular, that a MC on a background can have a complicated intensity profile due to a nonlinear superposition of pairs of bright and dark single solitons. PMID- 11031651 TI - Coupling of near-grazing microwave photons to surface plasmon polaritons via a dielectric grating AB - A dielectric grating on top of a planar metal substrate is shown to couple near grazing microwave photons to surface plasmon polaritons (SPPs). It is shown that when the grating grooves are oriented such that they are parallel to the plane of incidence (phi = 90 degrees), coupling to SPPs with both s- and p-polarized photons is possible at three different energies. It is demonstrated that one mode is coupled via p-polarized radiation and the other two modes are both coupled via s-polarized radiation. A multilayer, multishape differential grating theory allows the identities of each of the modes to be confirmed by modeling the electromagnetic fields above the metal substrate. In addition, a comparison between the experimentally derived reflectivity scans and the theoretical model is made. PMID- 11031652 TI - Interferometers and decoherence matrices AB - It is shown that the Lorentz group is the natural language for two-beam interferometers if there are no decoherence effects. This aspect of the interferometer can be translated into six-parameter representations of the Lorentz group, as in the case of polarization optics where there are two orthogonal components of one light beam. It is shown that there are groups of transformations which leave the coherency or density matrix invariant, and this symmetry property is formulated within the framework of Wigner's little groups. An additional mathematical apparatus is needed for the transition from a pure state to an impure state. Decoherence matrices are constructed for this process, and their properties are studied in detail. Experimental tests of this symmetry property are possible. PMID- 11031653 TI - Wave scattering from self-affine surfaces AB - Electromagnetic wave scattering from a perfectly reflecting self-affine surface is considered. Within the framework of the Kirchhoff approximation, we show that the scattering cross section can be exactly written as a function of the scattering angle via a centered symmetric Levy distribution for the general roughness amplitude, Hurst exponent and wavelength of the incident wave. Our prediction is supported by direct numerical simulations. PMID- 11031655 TI - Monte Carlo renormalization-group analysis of the lattice phi 4 model in D = 3,4 AB - We present a simple, sophisticated method to capture renormalization-group flow in Monte Carlo simulation, which provides important information of critical phenomena. We applied the method to the D = 3,4 lattice phi 4 model and obtained a renormalization flow diagram that well reproduces theoretically predicted behavior of the continuum phi 4 model. We also show that the method can be easily applied to much more complicated models, such as frustrated spin models. PMID- 11031654 TI - Localization of light in disordered dielectrics: an approach based on spectral statistics AB - We study numerically the fluctuation properties of the eigenvalues of the scalar wave equation in two dimensions for strong disorder. This equation mimicks properties of light in dielectrics. With increasing disorder, we find a transition from diffusive to localized behavior, in complete analogy to the case of Schrodinger waves (electrons). At low frequencies, we observe a suppression of disorder. This effect is caused by the wave number dependence of the disorder term in the wave equation, and has no analog in the case of electrons. PMID- 11031656 TI - Criticality and crossover in accessible regimes AB - The near-critical behavior of (d = 3)-dimensional Ising-model ferromagnets or simple lattice gases with equivalent first, second, and third nearest-neighbor interactions is studied through Monte Carlo simulations using histogram reweighting techniques and comparisons with series expansions. By carefully analyzing numerical data from relatively small finite systems using scaling and extrapolation methods, it is demonstrated that one can reliably estimate critical exponents, critical temperatures, and universal amplitude ratios, thereby distinguishing convincingly between different "nearby" universality classes and revealing systematic crossover effects. This study is preparatory to extending similar techniques to study criticality in continuum fluid models lacking symmetries, with Coulomb interactions, etc. PMID- 11031657 TI - Generalized variational principle for the time-dependent Hartree-Fock equations for a Slater determinant AB - The time-dependent Hartree-Fock equations are derived from a variational principle for the general N-body action of a Slater determinant of single electron orbitals. The variational principle generalizes commonly used variational treatments based on reduced two-body actions. The self-consistent field equations are found to contain time-dependent corrections to the standard mean-field interactions. Their physical significance is discussed and a time dependent phase shift to the Slater determinant is obtained that properly accounts for the total interaction energy in the mean-field approach. PMID- 11031658 TI - Library design in combinatorial chemistry by Monte Carlo methods. AB - Strategies for searching the space of variables in combinatorial chemistry experiments are presented, and a random energy model of combinatorial chemistry experiments is introduced. The search strategies, derived by analogy with the computer modeling technique of Monte Carlo, effectively search the variable space even in combinatorial chemistry experiments of only modest size. Efficient implementations of the library design and redesign strategies are feasible with current experimental capabilities. PMID- 11031659 TI - Nonsquare transfer-matrix technique applied to the simulation of electronic diffraction by a three-dimensional circular aperture AB - The transfer-matrix methodology is frequently used to deal with elastic scattering problems that require a solution of the Schrodinger or homogeneous Maxwell equations in the continuous part of their spectra. Until now, this technique was limited to representations associated with square transfer matrices. This paper extends the transfer-matrix methodology to enable consideration of general representations associated with nonsquare matrices. The theory is illustrated by the diffraction of a field-emitted electronic beam by a three-dimensional circular aperture. The application focuses on the dependence of the long-range angular spread on the aperture radius, by highlighting the effects of the field-emission tip shape and dimensions. PMID- 11031660 TI - Path-integral Monte Carlo simulations without the sign problem: multilevel blocking approach for effective actions AB - The multilevel blocking algorithm recently proposed as a possible solution to the sign problem in path-integral Monte Carlo simulations has been extended to systems with long-ranged interactions along the Trotter direction. As an application, results for the real-time quantum dynamics of the spin-boson model are presented. PMID- 11031661 TI - Solution of the quantum fluid dynamical equations with radial basis function interpolation AB - The paper proposes a numerical technique within the Lagrangian description for propagating the quantum fluid dynamical (QFD) equations in terms of the Madelung field variables R and S, which are connected to the wave function via the transformation psi = exp(R + iS)/[symbol: see text]. The technique rests on the QFD equations depending only on the form, not the magnitude, of the probability density rho = magnitude of psi 2 and on the structure of R = [symbol: see text]/2 ln rho generally being simpler and smoother than rho. The spatially smooth functions R and S are especially suitable for multivariate radial basis function interpolation to enable the implementation of a robust numerical scheme. Examples of two-dimensional model systems show that the method rivals, in both efficiency and accuracy, the split-operator and Chebychev expansion methods. The results on a three-dimensional model system indicates that the present method is superior to the existing ones, especially, for its low storage requirement and its uniform accuracy. The advantage of the new algorithm is expected to increase for higher dimensional systems to provide a practical computational tool. PMID- 11031663 TI - Model for correlations in stock markets AB - We propose a group model for correlations in stock markets. In the group model the markets are composed of several groups, within which the stock price fluctuations are correlated. The spectral properties of empirical correlation matrices reported recently are well understood from the model. It provides the connection between the spectral properties of the empirical correlation matrix and the structure of correlations in stock markets. PMID- 11031662 TI - Viability of competing field theories for the driven lattice gas AB - It has recently been suggested that the driven lattice gas should be described by an alternate field theory in the limit of infinite drive. We review the original and the alternate field theory, invoking several well-documented key features of the microscopics. Since the alternate field theory fails to reproduce these characteristics, we argue that it cannot serve as a viable description of the driven lattice gas. Recent results, for the critical exponents associated with this theory, are reanalyzed and shown to be incorrect. PMID- 11031664 TI - 1/f alpha fluctuations in a ricepile model AB - The temporal fluctuation of the average slope of a ricepile model is investigated. It is found that the power spectrum S(f) scales as 1/f alpha with alpha approximately 1.3 when grains of rice are added only to one end of the pile. If grains are randomly added to the pile, the power spectrum exhibits 1/f2 behavior. The profile fluctuations of the pile under different driving mechanisms are also discussed. PMID- 11031665 TI - Noise-aided control of chaotic dynamics in a logistic map AB - Controlling chaos involves employing small perturbations to a control parameter, resulting in the stabilization of the system (naturally chaotic) on one of the infinite unstable periodic orbits embedded in the chaotic attractor. In this Brief Report we study the constructive role of external noise in increasing the efficiency of controlling chaos. Using a logistic map as an example, control of chaotic dynamics is achieved using a linear delayed-feedback strategy. Working in the subthreshold regime of control (where the value of control constant is less than the minimum value required to stabilize the period-1 target state), system dynamics in the presence of superimposed noise (system plus control plus noise) exhibit a resonance effect. Furthermore it is observed that the time required to reach the target state decreases appreciably in the presence of an optimum level of noise. PMID- 11031666 TI - Multiple short-term memories in coupled weakly nonlinear map lattices AB - We investigate short-time memory storage in coupled map lattices with a periodic external input. In the case of linear coupled maps, the transient length necessary to achieve permanent memory is studied. We present numerical evidence that coupled weakly nonlinear maps are able to store multiple short-time memories, and use this fact to encode symbols in a matrix of pixels, using suitable control laws. PMID- 11031667 TI - Momentum distributions for the quantum delta-kicked rotor with decoherence AB - We report on the momentum distribution line shapes for the quantum delta-kicked rotor in the presence of environment induced decoherence. Experimental and numerical results are presented. In the experiment ultracold cesium atoms are subjected to a pulsed standing wave of near resonant light. Spontaneous scattering of photons destroys dynamical localization. For the scattering rates used in our experiment the momentum distribution shapes remain essentially exponential. PMID- 11031668 TI - Critical signal strength for effective decoding in diode laser chaotic optical communications AB - Synchronized external-cavity diode lasers are used for chaotic optical encryption and decryption. It is shown that effective decoding requires the signal strength to exceed a certain value determined by the precise operating conditions. PMID- 11031669 TI - Long periodic waves on an even beach AB - High-order Boussinesq-type equations for long waves over an even slope are derived and investigated. Potential and surface elevation for periodic wave motion are expanded in Fourier series up to the third harmonic inclusively. Coefficients of this series are expressed as polynomials of Bessel functions. PMID- 11031670 TI - Generalized Fourier law for heat flow in a fluid with a strong, nonuniform strain rate AB - We derive the leading terms of a generalized Fourier law for heat conduction in fluids under strong, nonuniform shear by expanding the heat flux vector as a Taylor series about the equilibrium state in powers of the temperature gradient, the velocity gradient (the first spatial derivative of the streaming velocity or the strain rate tensor), and, in an extension of previous work, the second spatial derivative of the streaming velocity (a third rank tensor). This results in a general macroscopic constitutive equation, independent of any microscopic model, and valid for all flow geometries. Assuming that the fluid is isotropic at equilibrium, we find a term representing heat flow due to a gradient in the square of the strain rate. This shows that it is possible for a nonuniform velocity gradient to generate a heat flow in the absence of a temperature gradient. We also find terms corresponding to heat flow parallel to the streamlines that are not present in uniform shear flow. PMID- 11031671 TI - Radial fluctuations and nonisotropic disclinations in nematic liquid crystals AB - The aim of this paper is to study the role of the radial symmetry in the profile of anisotropic disclinations of the nematic liquid crystals. It will be shown that when radial fluctuations are allowed a macroscopic term appears. This term preserves the known topology of these disclinations but changes their angular structure and the distribution of elastic energy. Furthermore, it is shown that one of the isotropiclike disclinations predicted by the usual approach is forbidden by the radial fluctuations. PMID- 11031672 TI - Force between two spherical inclusions in a nonlinear host medium AB - In an attempt to investigate the effect of nonlinear characteristics on particle interactions, a self-consistent mean-field theory in combination with a multiple image method has been employed to compute the interparticle force. Taking the nonlinearity of the host medium into account, the interparticle force exhibits a nonmonotonic behavior as the applied electric field is increased. We show that the interparticle force increases initially at low fields, goes through a maximum at an optimal electric field, then decreases with increasing field, and vanishes at a critical electric field at which the effective dielectric contrast between the host and the inclusions becomes zero. The influence of a larger volume fraction of inclusions on the interparticle force is also investigated. PMID- 11031673 TI - Finite resolution effects in the analysis of the scaling behavior of rough surfaces AB - We investigate the influence of finite spatial resolution in the analysis of the scaling behavior of rough surfaces. We analyze such an effect for two usual measurement methods: the local width and the height-height correlation function. We show that while the correlation function is insensitive to finite resolution effects for practical purposes, the local width presents correction terms to the scaling law, leading to an effective value of the local roughness exponent smaller than the theoretically expected. We also show that a functional scaling relation can only be properly formulated in terms of the height-height correlation function. PMID- 11031674 TI - Fluid-fluid transitions of hard spheres with a very-short-range attraction AB - Hard spheres with an attraction of range a tenth to a hundredth of the sphere diameter are constrained to remain fluid even at densities when monodisperse particles at equilibrium would have crystallized, in order to compare with experimental systems that remain fluid. They are found to have a fluid-fluid transition at high density. As the range of the attraction tends to zero, the density at the critical point tends toward the random-close-packing density of hard spheres. PMID- 11031675 TI - Reinterpretation of a Kardar-Parisi-Zhang equation-based classification AB - The velocity versus tilt behavior, on which the Kardar-Parisi-Zhang (KPZ) equation-based quenched Edward-Wilkinson-directed percolation depinning (QEW-DPD) classification scheme of models of rough interface growth in a medium with quenched disorder is based, is reinterpreted. The consideration of the screen grid of pixels in computer simulation interface propagation allows an explanation of tilt-velocity behavior without assuming vanishing or divergence in the depinning transition of the KPZ parameter lambda which is found to have a unique, measurable value in DPD. Random field Ising model-like velocity-tilt behavior in the QEW class is shown to either correspond to zero lambda. The possibility of obtaining low velocity v(m) behavior from the KPZ method raises some interesting questions about the depinning transition. PMID- 11031676 TI - Correlated patterns in nonmonotonic graded-response perceptrons AB - The optimal capacity of graded-response perceptrons storing biased and spatially correlated patterns with nonmonotonic input-output relations is studied. It is shown that only the structure of the output patterns is important for the overall performance of the perceptrons. PMID- 11031677 TI - [Decision making concerning very old patients with deteriorating health living at home]. AB - A woman aged 87 and three men aged 87, 86 and 83, respectively, lived at home or in an old-people's home, in poor physical condition due to old age and diseases such as COPD and heart failure. One man was demented, the others did not wish hospital admission in case of further deterioration. When the condition worsened due to infections and fractures, the GP to a limited extent performed diagnostics and treatment. The four patients died, three of them after admission and intensive treatment because the restricted policy had not been communicated clearly. In decision-making about the management of geriatric patients, it is important that the GP knows the patient's wishes, correctly assesses his physical condition and prognosis and communicates well with the patient's next of kin and other care-givers. PMID- 11031678 TI - [Roaming through methodology. XXIV. Problems in generalizing from randomized intervention studies with selected patient populations]. AB - The generalizability of intervention research is dependent of the degree of success in including patients who represent the disease category for which the intervention might be considered. To be relevant for medical practice, estimates of intervention effects need to be quantitatively generalizable. This requires specification of the characteristics of the intervention (e.g. dosage and timing) and of the disease outcome, and of socalled effect-modifying factors: disease severity, co-interventions and the characteristics of the patients which influence the magnitude of the intervention effect. Literature remains less clear about the necessity to specify these effect-modifying factors. If effect modification is seriously considered, the investigator must decide either to design the trial in such a way that the subgroup-specific effects may be estimated, or to restrict the study to one homogeneous stratum of patients. Despite many theoretical reasons and some empirical examples of substantial effect modification by characteristics of patients, systematic comparison of randomised (selected populations) and non-randomised studies (less selection) fail to show many differences. This suggests that differences between subgroups are not very frequent and that trial results, provided they are adequately specified, generally do have a high degree of generalizability. PMID- 11031679 TI - [Summary of the guideline 'Sentinel node biopsy in breast cancer.' Dutch Work Group 'Sentinel Node Biopsy for Breast Cancer']. AB - The team Sentinel Node Biopsy Breast Cancer issued guidelines for the performance of the sentinel node procedure. The team took into account the problems resulting from the introduction of the technique. The eligible group consists mainly of women with demonstrated breast cancer with a diameter of < or = 2 cm without palpable axillary nodes. Contraindications include presence of more than one tumour, pathological nodes in the ipsilateral axilla and previous treatment of breast or axilla. After injection of a radioactive substance, the sentinel node is detected during the operation using a gamma probe. The tracer is injected 2 to 24 hours before the operation in or near the tumour. Shortly before the incision, patent blue is injected as well, improving the detectability of the sentinel node. One or several sentinel nodes are examined histologically. The pathology report meets minimal requirements. In the planning and introduction of the technique in a hospital a locally responsible radiation expert should be involved. The procedure should be performed a number of times and evaluated with reference to a histopathological result before it is decided to draw clinical conclusions from the procedure. For the necessary evaluations of the procedure central registration is an absolute condition. PMID- 11031680 TI - [Diagnostic image (4)(Bronchoalveolar lavage fluid)]. AB - Two apparently healthy male volunteers, 28 and 32 years old, were examined before participation in a drug trial. Bronchial fluid was obtained. The lavage fluid from the second volunteer who had smoked 20 cigarettes a day for the past 16 years showed marked brownish discolouration, as compared to the transparent fluid of the non smoking first volunteer. PMID- 11031682 TI - [Severe obstipation due to eating unshelled sunflower seeds]. AB - In two boys aged 13 and 6 years respectively obstipation and a paradoxical diarrhoea developed four days after consumption of unshelled sunflower seeds. The cause was impaction of the unshelled sunflower seeds in the rectum. Both patients required general anaesthesia to remove the sunflower seeds via proctoscopy. It is known that eating unshelled sunflower seeds can cause impaction of the shells in the rectum (causing a bezoar). Removing the seeds under general anaesthesia is practically always necessary. Considering the serious symptoms and the invasive consequences which are not without risks, parents and children should be warned about the danger of eating unshelled sunflower seeds. PMID- 11031681 TI - [Aortocoronary bypass surgery; at least 6 months follow-up required for assessment of postoperative course]. AB - OBJECTIVE: To determine how the mortality and morbidity after aortocoronary bypass surgery during the hospital stay correspond with those in the first 6 months after the operation. DESIGN: Prospective and retrospective. METHOD: Data of 563 patients undergoing isolated myocardial revascularization in 1998 in the Academic Hospital Nijmegen, the Netherlands, were collected on mortality and morbidity 180 days after the operation. The Parsonnet score was used to assess the operative risk of the patients. RESULTS: The patient group consisted of 422 males and 141 females, with a mean age of 64.3 years. The median Parsonnet score was 6 (range: 0-50). The mean hospital stay was 7.9 days (SD: 9.4). The hospital mortality was 3.9% (22/563). The follow-up was 100%. During the follow-up 11 patients died, 10 from cardiac causes. Six months postoperatively the mortality was 5.9% (22/563). The Kaplan-Meier curve of the survival probability rate showed a persistent decrease, mainly in the higher risk patients. Non-fatal cardiovascular events were registered in 9.6% of the surviving patients (51/530). CONCLUSION: The hospital mortality and morbidity after aortocoronary bypass surgery were considerably lower than the total postoperative mortality and morbidity during the first 6 months after the operation. For assessment of the postoperative course, systematic follow-up over 6 months is necessary. PMID- 11031683 TI - ['Another breakthrough': general news reports raise false hopes]. AB - Recently a breakthrough in research was reported in the Dutch press. The results were considered of major importance for the treatment of patients with Parkinson's disease, motor neuron disease and Alzheimer's disease. Thereafter societies of patients and neurologists were overloaded with questions. They had to explain again and again that false expectations had been aroused. The road from basic research to a proven beneficial therapy is long. Promising results are at many phases of the development of a treatment at risk of disappearing. Reports from the laboratory on new treatments are always too early. In order to prevent false expectations, submissions to the press ought to be based on firm evidence as is usual with publications in professional journals, editors of the press should scrutinize the claims made by researchers, and, in addition, the general public should be educated on the diagnostic process and the assessment of treatment efficacy. PMID- 11031684 TI - [Travel experiences in Central and Eastern Europe: Romania: the general's kisses]. AB - The public face of Rumania is characterized by extreme contrasts between very poor and very rich people. The state is near bankruptcy, industry is slow in recovering from many years of madcap rule: politics, tourism and agriculture are in chaos, doctors are seriously underpaid and good health care is only available to the newly rich. World Bank loans have been largely spent on sophisticated hospital instrumentation, but the primary health care barely manages to survive. The capital has seriously suffered from Ceausescu's megalomania and many historic cities show the remnants of past glory. The country sometimes resembles a disorganized beehive. Rumanians try to steer a middle course between bouts of self-pity and their intense wish to become full members of the European Union. They are not averse to play-acting and one should not be surprised to receive a cordial accolade from a medical professor who chairs government committee meetings in full general's attire. PMID- 11031685 TI - [Human papillomavirus and etiology of cervical cancer: concept of carcinogenesis]. PMID- 11031686 TI - American Paraplegia Society 46th annual conference. Abstracts. PMID- 11031687 TI - International Society for the Study of the Origin of Life (ISSOL) meeting. July 11-16, 1999, San Diego, California, USA. Abstracts. PMID- 11031688 TI - [Ultrasonography 2000. 24th Three-country meeting of the Swiss, German, and Austrian Societies for Ultrasound in Medicine. 7-9 September 2000, Vienna. Abstracts]. PMID- 11031689 TI - Abortive rabies virus central nervous infection is controlled by T lymphocyte local recruitment and induction of apoptosis. AB - Nonfatal paralysis, induced by the attenuated Pasteur strain of rabies virus, is characterised by local and irreversible flaccid paralysis of the inoculated limbs. We characterised the spread and localisation of virus in the CNS of infected mice, determined the nature of cell injury and examined the role of the immune response. Data indicate that infection of BALB/c mice induced paralysis in 60% of infected mice, the others recovering without sequelae. In both groups of mice, virus was detected in restricted sub-populations of neurons from the brain and spinal cord, and intensity of the neuropathology correlated with levels of rabies RNA and apoptotic infected neurons. However, apoptosis of neurons and paralysis were not due to a direct deleterious effect of the virus, but induced by a T-dependent immune response, as evidenced by their absence in nude mice. Paralysed and asymptomatic mice developed a similar rabies virus-specific IgG2a antibody response, thus excluding the role of any modification of the humoral immune response. In contrast, three events were critically associated with the development of neurological symptoms: the amount of virus in the CNS, the level of apoptosis in both infected neurons and uninfected surrounding cells and the progressive parenchymal infiltration of CD4+ and CD8+ T cells at the site of infection. These data suggest that during nonfatal rabies infection, the levels of viral replication and primary degeneration of infected neurons by apoptosis could be responsible for the infiltration of T lymphocytes capable of inducing secondary degeneration of neural cells. PMID- 11031690 TI - Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetics approach. AB - The rabies virus glycoprotein (G) gene of the highly neuroinvasive and neurotropic strains SHBRV-18, CVS-N2c, and CVS-B2c was introduced into the non neuroinvasive and less neurotropic SN-10 strain to provide further insight into the role of G in the pathogenesis of rabies. Phenotypic analyses of the recombinant viruses revealed, as expected, that the neurotropism of a particular rabies virus strain was a function of its G. Nevertheless, the pathogenicity of the recombinant viruses was, in every case, markedly lower than that of the wild type viruses suggesting that while the G dictates neurotropism, other viral attributes are also important in pathogenesis. The low pathogenicity of the recombinant viruses is at least in part due to a strong increase in transcription activity. On the other hand, the production of infectious virus by the R-SHB18 recombinant virus-infected cells was significantly delayed by comparison with SHBRV-18 wild-type virus infected-cells. Replacement of the R-SHB18 G cytoplasmic domain, transmembrane domain, and stem region with its SN-10 G counterparts neither results in a significant increase in budding efficiency nor an increase in pathogenicity. These results suggest that an optimal match of the cytoplasmic domain of G with the matrix protein may not be sufficient for maximal virus budding efficiency, which is evidently a major factor of virus pathogenicity. Our studies indicate that to maintain pathogenicity, the interactions between various structural elements of rabies virus must be highly conserved and the expression of viral proteins, in particular the G protein, must be strictly controlled. PMID- 11031691 TI - Interactions between HIV-infected monocyte-derived macrophages and human brain microvascular endothelial cells result in increased expression of CC chemokines. AB - The presence of perivascular monocytic infiltration is a major hallmark of HIV-1 associated dementia. Since CC chemokines are chemoattractant cytokines that are able to attract T cells and monocytes/macrophages to sites of inflammation, and since infiltrating monocytes/macrophages remain in close contact with the brain endothelium, we investigated whether interactions between HIV-1-infected macrophages and brain endothelium result in an altered chemokine production. We found an increased mRNA expression of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1 alpha and MIP-1 beta, and RANTES by macrophages after HIV-1 infection. Interactions between HIV-infected macrophages and brain microvascular endothelial cells resulted in an additional upregulation of chemokine mRNA expression, during cell-cell contact as well as in a trans-well system. Since IL-1 beta can function as a modulator of chemokine expression we investigated if interleukin-1 beta could be involved in the regulation of chemokine induction. Coculturing of HIV-infected macrophages and endothelial cells resulted in immune-activation as indicated by increased mRNA expression of IL-1 beta. Subsequently, addition of a neutralizing antibody against IL-1 beta resulted in altered chemokine expression by macrophages, but not by endothelial cells. Thus, IL-1 beta appears to play a major role in the regulation of chemokines during cellular interactions in HIV-associated dementia, but other factors may also be involved. PMID- 11031692 TI - Cerebrospinal fluid viral load is related to cortical atrophy and not to intracerebral calcifications in children with symptomatic HIV disease. AB - The relationships between viral load in plasma and cerebrospinal fluid (CSF) and computed tomography (CT) brain scan abnormalities were studied in 39 children between 0.5 and 13 years of age with symptomatic HIV-1 disease. Quantitative RNA PCR was used to determine HIV-1 RNA levels and a semiquantitative analog rating technique was used to evaluate non-contrast CT brain scans. CSF HIV-1 RNA copy number correlated significantly with CT brain scan ratings for severity of cortical atrophy (r = 0.36; P < 0.05) but not with ratings of intracerebral calcifications (r = -12; NS). The difference between these two correlations was significant (P < 0.05). Plasma HIV-1 RNA copy number did not correlate significantly with any CT brain scan ratings or with CSF viral load (r = 0.05; NS). Severity of cortical atrophy appeared to reflect the level of viral load in the CSF, supporting the notion that active HIV-1 replication in the CNS is at least in part responsible for such brain abnormalities in children. The lack of correlation of intracerebral calcifications with other CT brain scan abnormalities as well as with CSF viral load suggests that this lesion is relatively independent and may reflect a different neuropathologic process. PMID- 11031693 TI - Analysis of the transcriptional control region in progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease caused by the human polyomavirus JCV. The hypervariable noncoding transcriptional control region (TCR) largely regulates replication of JCV in glial cells. Two distinct types of the TCR can be distinguished. Type II is derived from the archetype sequence. All type I TCRs, including the prototypical Mad-1 isolate contain a 23 bp deletion at nucleotide position 36. In a prospective study, TCR DNA could be amplified and sequenced in 16/29 (55%) suspect cases of PML from the cerebrospinal fluid (CSF) and in 14/28 (50%) urine samples. Sequencing of the CSF TCR identified Mad-1 like sequences in 5/17 (29.5%) instances and a type II TCR in 12/17 (70.5%) of cases. Of 14 urine TCRs, 12 (86%) displayed the archetype sequence, while two showed complex rearrangements. In all type II TCR sequences, the tst-1/oct-6 binding sites present in regions C and E of Mad-1 were missing. In 11/12 type II TCR sequences the pentanucleotide repeat in region A showed a G to T substitution of one nucleotide at position 36 relative to the Mad-1 TCR. All type II TCRs contained an Sp1 binding site at the beginning of region B. Of the 12 TCR type II sequences, 10 (83%) were of the 'D-retaining' pattern. In eight of these (80%) additional juxtapositioned nuclear factor 1, glial factor 1 and/or AP 1 binding motifs were created by duplications and/or insertions in region D. These findings indicate that type II TCRs are frequently present in PML and suggest to use TCR type II constructs for in vitro and in vivo studies of the evaluation of the functional role of DNA binding motifs. PMID- 11031695 TI - Chronic parvovirus B-19 meningoencephalitis with additional detection of Epstein Barr virus DNA in the cerebrospinal fluid of an immunocompetent patient. AB - Parvovirus B19 DNA was detected by polymerase chain reaction in the brain biopsy specimen from a 67-year-old immunocompetent woman with severe chronic lymphocytic meningoencephalitis. In addition to parvovirus B19, Epstein-Barr virus DNA was identified in the CSF. Genomic material from Epstein-Barr virus was absent in the brain tissue. Clinical symptoms and CSF pleocytosis improved under long-term corticosteroid-treatment. The aetio-pathogenetic role of parvovirus B19 and the possible meaning of the additionally detected Epstein-Barr virus DNA are discussed. PMID- 11031694 TI - A single tube PCR assay for simultaneous amplification of HSV-1/-2, VZV, CMV, HHV 6A/-6B, and EBV DNAs in cerebrospinal fluid from patients with virus-related neurological diseases. AB - Cerebrospinal fluid (CSF) specimens from 27 patients with encephalitis, meningitis, and other neurological diseases were studied for the presence of herpes simplex virus types 1 and 2 (HSV-1/-2), varicella-zoster virus (VZV), cytomegalovirus (CMV), human herpesviruses 6A and 6B (HHV-6A/-6B) and Epstein Barr virus (EBV) DNA using the polymerase chain reaction (PCR) method. The DNAs were amplified using two sets of consensus primer pairs in a single tube, bringing simultaneous amplification of the herpesviruses. The PCR products were analyzed by agarose gel electrophoresis, and Southern blot hybridization with virus-type specific probes, thus allowing discrimination between the different types of herpesviruses to be made. Each virus-specific probe was highly specific for identifying the PCR product. Thirty CSF specimens from 13 patients with encephalitis and 10 specimens from 10 patients with meningitis, respectively, were examined using this method. Eight patients with encephalitis and six with meningitis were positive for different herpesviruses, including patients with coinfections (HSV-1/-2 and VZV, VZV and CMV). Among four CSF specimens from four patients with other neurological disorders, dual amplification of CMV and EBV was present. Since identification of the types of herpesviruses in this system requires a very small amount of CSF, and is completed with one PCR, it is useful for routine diagnosis of herpesvirus infections in diagnostic laboratories. The viruses responsible for central nervous system infection are easily detected with various coinfection and serial patterns of herpesviruses, by this consensus primer-based PCR method. This may give an insight into the relationship between virus-related neurological diseases (VRNDS) and herpesvirus infections. PMID- 11031696 TI - HELLP syndrome. AB - HELLP syndrome is a serious, life-threatening form of pre-eclampsia with a typical laboratory triad. The incidence of the disease is reported as being 0.17 0.85% of all live births. There has been, to date, neither reliable early recognition nor effective prevention of HELLP syndrome. As a result of endothelial dysfunction, activation of intravascular coagulation occurs with fibrin deposition in the capillaries and consecutive microcirculation disorders. The disease manifests itself on average between 32-34 weeks' gestation. HELLP syndrome will occur postpartum in up to 30% of the cases. The clinical cardinal symptom of the disease is right upper quadrant pain or epigastric pain accompanied with nausea, vomiting and malaise. In 20% of the cases with HELLP syndrome there is no hypertension and 5-15% of the pregnant patients present a low level of proteinuria or none at all. The early recognition of hemolysis is most sensitively managed by the determination of the serum haptoglobin. The increase of the aspartate transaminase (AST) and the alanine transaminase (ALT) often precedes a decrease in platelets. The course of HELLP syndrome is incalculable. It is universally agreed that a pregnancy from 32-34 weeks should be immediately delivered. Before 32-34 weeks, expectant management is generally possible in a perinatal center. The frequency for a repeated hypertensive disease in pregnancy ranges from 27% to 48%. PMID- 11031697 TI - Perinatal brain injury. AB - Perinatal brain damage in the mature fetus is usually brought about by severe intrauterine asphyxia following an acute reduction of the uterine or umbilical circulation. The areas most heavily affected are the parasagittal region of the cerebral cortex and the basal ganglia. The fetus reacts to a severe lack of oxygen with activation of the sympathetic-adrenergic nervous system and a redistribution of cardiac output in favor of the central organs (brain, heart and adrenals). If the asphyxic insult persists, the fetus is unable to maintain circulatory centralization, and the cardiac output and extent of cerebral perfusion fall. Owing to the acute reduction in oxygen supply, oxidative phosphorylation in the brain comes to a standstill. The Na+/K+ pump at the cell membrane has no more energy to maintain the ionic gradients. In the absence of a membrane potential, large amounts of calcium ions flow through the voltage dependent ion channels, down an extreme extra-/intracellular concentration gradient, into the cell. Current research suggests that the excessive increase in levels of intracellular calcium, so-called calcium overload, leads to cell damage through the activation of proteases, lipases and endonucleases. During ischemia, besides the influx of calcium ions into the cells via voltage-dependent calcium channels, more calcium enters the cells through glutamate-regulated ion channels. Glutamate, an excitatory neurotransmitter, is released from presynaptic vesicles during ischemia following anoxic cell depolarization. The acute lack of cellular energy arising during ischemia induces almost complete inhibition of cerebral protein biosynthesis. Once the ischemic period is over, protein biosynthesis returns to preischemic levels in non-vulnerable regions of the brain, while in more vulnerable areas it remains inhibited. The inhibition of protein synthesis, therefore, appears to be an early indicator of subsequent neuronal cell death. A second wave of neuronal cell damage occurs during the reperfusion phase. This cell damage is thought to be caused by the postischemic release of oxygen radicals, synthesis of nitric oxide (NO), inflammatory reactions and an imbalance between the excitatory and inhibitory neurotransmitter systems. Part of the secondary neuronal cell damage may be caused by induction of a kind of cellular suicide programme known as apoptosis. Interestingly, there is increasing evidence from recent clinical studies that perinatal brain damage is closely associated with ascending intrauterine infection before or during birth. However, a major part of this damage is likely to be of hypoxic-ischemic nature due to LPS-induced effects on fetal cerebral circulation. Knowledge of these pathophysiological mechanisms has enabled scientists to develop new therapeutic strategies with successful results in animal experiments. The potential of such therapies is discussed here, particularly the promising effects of intravenous administration of magnesium or postischemic induction of cerebral hypothermia. PMID- 11031698 TI - An overview of the possible relationship between antenatal pharmacologic magnesium and cerebral palsy. AB - Magnesium sulfate is currently being used in obstetric practice for either eclamptic seizure prophylaxis or for tocolysis, in some countries. Evidence for its use in preeclampsia is credible, whereas the evidence for its use as a tocolytic is limited, if not absent. Of interest, the findings of two epidemiologic studies have suggested a third possible use for antenatal pharmacologic magnesium sulfate, namely, as a neuroprotectant against the later development of cerebral palsy in the newborn. In support of this hypothesis are laboratory data, much of which have to do with the modulation of cellular membrane receptors. Unfortunately, during the Magnesium and Neurologic Endpoints Trial (MagNET), while attempting to confirm the neuroprotective effect of magnesium sulfate, the occurrence of excess total pediatric mortality in those children exposed to magnesium led to early termination of the trial. Nonetheless, despite the alarming findings in MagNET, it is conceivable that exposures to doses of magnesium sulfate less than those often used for aggressive tocolysis may be neuroprotective without being lethal. Other randomized controlled trials now underway may answer this important question. PMID- 11031699 TI - A corticotropin releasing hormone receptor antagonist does not delay parturition in rats. AB - OBJECTIVE: To determine the effects of CP-154, 526, a corticotropin releasing hormone (CRH) receptor antagonist, on the length of normal rat gestation. STUDY DESIGN: Twenty-four timed-pregnant Sprague-Dawley rats were purchased for this study. The drug and placebo were administered to the animals using an osmotic pump surgically inserted in the dorsal subcutaneous space. Six animals received 6 mg/kg/day of the drug, six animals received 12 mg/kg/day of the drug and twelve animals received the placebo. The gestational period, weight of each pup and number of pups in each litter were recorded and compared in the drug group versus placebo group. RESULTS: No difference was noted in the gestational period of the drug and placebo rats. The mean weight of pups in both the drug and placebo groups was 6.18 g. The number of pups per litter were similar in the drug and placebo groups. CONCLUSION: Antagonism of CRH receptors in rats has no effect on the length of gestational period, pup weight or number of pups per litter. Further studies are needed to define the role of CRH and its antagonism in primate pregnancy, as has been done in sheep. PMID- 11031700 TI - Clinical usefulness of pulse oximetry in the fetus with non-reassuring heart rate pattern? AB - The objective of this study was the evaluation of intrapartum pulse oximetry as an indicator of fetal distress and the condition of the newborn during clinical routine surveillance in an University Perinatal Center. Between 1998 and 1999 pulse oximetry (SpO2) was used additionally to routine fetal monitoring by electronic fetal heart rate tracing (CTG) and fetal blood sampling (FBA) in 128 cases with nonreassuring heart rate pattern. Cut off values were FIGO Score < 8 for the heart rate pattern and for fetal blood sampling during labor results of < 7.25 (preacidosis). The condition of the newborn was defined by the APGAR score with the cut off < 7 at 1 minute, while the biochemical status was evaluated by means of arterial blood sampling of the umbilical artery directly after birth using a pH of < 7.20 to verify acidosis. Predictive values of critically low SpO2 values (< 30%) for at least 10 minutes as well as corresponding sensitivities and specificities were calculated together with 95% confidence intervals to identify fetal distress or a depressed condition of the newborns. Of 128 fetuses included in this study 66 (52%) were born spontaneously, 23 (18%) were born by operative vaginal delivery and 39 (31%) by means of cesarean section. The high rate of cesarean section was due to cephalopelvic disproportion in 29 cases. Fetal outcome was evaluated with a clinical score: mean APGAR score value 8.5 SD +/- 1. The mean value of the pH in the umbilical artery was 7.23 +/- 0.04. During a SpO2 monitoring period of 18,381 minutes we analyzed a contact time of 63%. Comparing SpO2 values of < 30% with preacidosis in the fetal blood sampling, we found a positive predictive value of merely 0.17 (95% CI: 0.00-0.64). Of 9 preacidotic cases during delivery only 1 was indicated by a saturation value below 30% (sensitivity 0.11, 95% CI: 0.00-0.48). The specificity and negative predictive value were calculated as 0.83 (95% CI: 0.65-0.94) and 0.76 (95% CI: 0.58-0.89) respectively. Of eleven cases with acidosis in the blood of the umbilical cord artery, pH < 7.20, only 2 were indicated by a SpO2 values below 30%. Which is equivalent to a sensitivity of 0.18 (95% CI: 0.03-0.52). Results of a receiver operator curve analysis showed no substantial deviation from the diagonal. The area under the curve was 0.62, the 95% CI (0.47-0.76) indicating no significant discrimination. Three of 49 fetuses with SpO2 recording during the last 10 minutes were born in clinical depressed status (APGAR < 7). None was indicated by a SpO2 value below 30%. CONCLUSION: Fetal distress and impaired condition of the newborn are not identified or predicted during routine application of SpO2 monitoring in the fetus during labor with adequate safety. PMID- 11031701 TI - Responses to vibroacoustic stimulation in a fetus with an encephalocele compared to responses of normal fetuses. AB - BACKGROUND: Observation of fetal movement and fetal heart rate (FHR) responses to repeated vibroacoustic stimulation (VAS) might be useful as a measure to assess fetal well-being and to assess the integrity of the fetal central nervous system (CNS). We observed the movement and FHR responses to repeated VAS of a term fetus with a serious brain anomaly as compared to responses of normal term fetuses. SUBJECTS, METHODS, RESULTS: In 37 normal term fetuses and in a term fetus with an encephalocele we studied movement and FHR response to repeated VAS. All normal fetuses responded within 1 s after stimulation with general body movement and FHR acceleration. At 36 gestational weeks, no movement or FHR responses were seen in the fetus with an encephalocele. Repetition of the test in this fetus after one week still showed no response to repeated VAS. CONCLUSION: Normal fetuses showed movement and FHR responses to external stimulation. The fetus with an encephalocele did not respond to repeated VAS with a movement or FHR acceleration. Case studies in fetuses with structural anomalies of the CNS are needed to gain insight into the spectrum of possible responses to VAS. PMID- 11031702 TI - Neonatal sepsis of vertical transmission: an epidemiological study from the "Grupo de Hospitales Castrillo". AB - A prospective multicenter study was designed to assess the epidemiology of neonatal sepsis of vertical transmission in Spain. The study was carried out by the "Grupo de Hospitales Castrillo" that included the neonatal services of 19 tertiary care (reference) hospitals and 9 secondary care hospitals. Prospective data from infants with culture-proved neonatal sepsis, clinical sepsis and bacteremia were recorded for 1995 to 1997. In a total of 203,288 neonates, proven sepsis was diagnosed in 515 (rate of 2.5 per 1000 live births), clinical sepsis in 724 (rate of 3.6 per 1000 live births), and bacteremia of vertical transmission in 155 (rate of 0.76 per 1000 live births). Very low birth weight (VLBW) infants (< or = 1500 g) showed a significantly higher incidence of confirmed sepsis (26.5 per 1000 live births) and clinical sepsis (32.4 per 1000 live births) than infants weighing > 1500 g. Streptococcus agalactiae was the most frequent causative pathogen in cases of proven sepsis (51%) and bacteremia (33%), but Escherichia coli was the most frequently recovered organism in the VLBW group. The mortality rate of proven sepsis was significantly higher than that of clinical sepsis (8.7% versus 4.3%) (P < 0.01). In the VLBW cohort, there were no significant differences in the mortality rate between proven sepsis and clinical sepsis. In conclusion, clinical sepsis was the most frequent diagnosis, probably related to intrapartum chemoprophylaxis. Streptococcus agalactiae was the most frequent causative pathogen of culture-positive sepsis and bacteremia, whereas E. coli was the most significant in VLBW infants. PMID- 11031703 TI - Why are cesarean delivery rates so high in diabetic pregnancies? AB - AIMS: The purpose of this study was to examine factors relevant to mode of delivery in term pregnancies complicated by gestational and pre-gestational diabetes. METHODS: A retrospective chart review of term (> or = 37 weeks) singleton pregnancies complicated by Class A2 through Class R pregnancies which delivered from 1991-1997 was performed. Exclusion criteria were prior cesarean delivery, non-vertex presentation, fetal structural defects, or any contraindications to vaginal delivery. Maternal and fetal factors relevant to mode of delivery were examined and compared. Stepwise logistic regression analysis was performed to examine factors predictive of delivery mode. RESULTS: A total of 148 patients met study criteria. Induction rates were 60.9% for gestational and 79.8% for pre-gestational diabetics. The overall cesarean delivery rate by Diabetes Class for A2, B, C, D-F pregnancies was 20.3%, 40%, 37%, and 57.1% respectively. In Class A2 pregnancies no factor was associated with cesarean delivery and only nulliparity (p = 0.03) was associated in Class B F pregnancies. CONCLUSIONS: These results suggest that physician factors may play an important role in the risk for cesarean delivery in our diabetic population. PMID- 11031704 TI - Uterine artery pseudo-aneurysm: diagnosis and therapy during pregnancy. AB - Arterial aneurysms and pseudo-aneurysms are a rare but recognized cause of obstetric hemorrhage. Diagnosis during pregnancy, prior to rupture, is exceptional. We report the first case of diagnosis and treatment of an uterine artery pseudo-aneurysm during pregnancy. PMID- 11031705 TI - Minimally invasive endoscopy in the treatment of preterm premature rupture of membranes by application of fibrin sealant. AB - We report only the 3rd case of closure of amniorrhexis following genetic amniocentesis. Our technique is the first to use endoscopic visualization of the rupture site and apply maternal platelets and fibrinogen/thrombin (Hemaseel Haemacure Corp Sarasota F1). The patient underwent repair at 20.6 weeks, 26 days after spontaneous rupture of membranes post-amniocentesis. At the time of the procedure the amniotic fluid index was 1 cm. Patient was delivered at 32.3 weeks secondary to complications of diabetes and severe preeclampsia. The neonate had APGARS of 7 at 1 min and 8 at 5 min and was discharged home on Day 21 of life. PMID- 11031706 TI - Carl Ludwig Award 2000: experimental glomerulonephritis: the use of genetically deficient mice to elucidate the role of leukocytes and leukocyte adhesion receptors. PMID- 11031707 TI - Hans U. Zollinger Award 2000: problems from the clinic, answers from the laboratory: mutual interactions that lead to rationales for an optimized and individualized immunosuppressive regimen. PMID- 11031708 TI - Franz Volhard Award 2000: angiotensin II signalling in the podocyte. PMID- 11031709 TI - Signalling pathways of nitric oxide. PMID- 11031710 TI - Megalin/GP330 and pathogenetic concepts of membranous glomerulopathy (MGN). PMID- 11031711 TI - Proteinuria and progression of chronic renal disease. PMID- 11031712 TI - Apoptosis in resolution of inflammation. PMID- 11031713 TI - Co-stimulatory signals during recognition of allo-antigens. PMID- 11031714 TI - Vasopeptidase inhibition as a new concept in antihypertensive therapy. PMID- 11031715 TI - New parameters in kidney biopsy diagnostic-morphometry. PMID- 11031716 TI - Molecular pathophysiology of renal calcium handling. PMID- 11031717 TI - Genetic variation of the renin system--effects on blood pressure and the kidney. PMID- 11031718 TI - Organ donation and justice in organ allocation. PMID- 11031719 TI - New developments in clinical immunosuppression. AB - Over the last few years, the armamentarium of immunosuppressants has been enriched by a variety of new drugs, that have helped to further reduce the incidence of acute rejection episodes after kidney transplantation. Nevertheless the search for new compounds drugs continues, until tolerance, the ultimate goal of transplantation medicine can be achieved. Meanwhile we should appreciate that neither all drugs nor all patients are equal and tailor the immunosuppression to the patients' needs. To define patient groups who benefit most from different immunosuppressive protocols especially in regard to long term allograft and patient survival will warrant further prospective controlled trials. PMID- 11031720 TI - Recent advances in the understanding of water metabolism in heart failure (involvement of aquaporin). PMID- 11031721 TI - Role of matrix metalloproteinases in mesangial proliferative glomerulonephritis. PMID- 11031722 TI - Smoking--a risk factor for progression of renal disease. PMID- 11031723 TI - Therapy of patients with IgA-nephropathy: a critical appraisal. PMID- 11031724 TI - The role of thrombospondin in renal disease. PMID- 11031725 TI - Determination of malondialdehyde in biological samples by solid-phase extraction and high-performance liquid chromatography. AB - An analytical procedure for determination of malondialdehyde in tissue homogenates and blood serum was developed. A reaction with 2,4 dinitrophenylhydrazine is used followed by cleaning up of the derivative by solid phase extraction. The samples were analyzed by isocratic high-performance liquid chromatography (HPLC) using a narrow-bore HPLC-column. A good separation of the 1 pyrazole peak from that of 2,4-dinitrophenylhydrazine was observed. A high linear dependence was established by the concentration of 1-pyrazole in the range of 10 5000 ng/ml. The detection limit of the method applied for tissue homogenates and blood serum was approximately 10 ng/ml or lower, and RSD of the method was 9% (n = 8). The peak of 1-pyrazole for these samples was well separated from the other matrix peaks. Experiments carried out evaluated that the solid-phase extraction might be an effective step of the sample preparation, significantly increasing the selectivity of the analysis and the life-time of the column. The method seems to be applicable for determination of malondialdehyde in different biological samples. PMID- 11031726 TI - In vitro antioxidant activity of piperine. AB - Oxygen radical injury and lipid peroxidation have been suggested as major causes of atherosclerosis, cancer, liver disease and the aging process. Piperine, having an antiinflammatory effect, has been demonstrated in in vitro experiments to protect against oxidative damage by inhibiting or quenching free radicals and reactive oxygen species and hydroxyl radicals. The effect on lipid peroxidation was also examined and IC50 values were calculated. Piperine was found to act as a hydroxyl radical scavenger at low concentrations, but at higher concentrations, it activated the fenton reaction resulting in increased generation of hydroxyl radicals. Whereas it acts as a powerful superoxide scavenger and IC50 is 1.82 mM, a 52% inhibition of lipid peroxidation was observed at a dose of 1400 microM with an IC50 of 1.23 mM. The results depict that piperine possesses direct antioxidant activity against various free radicals. This study also opens newer views on the potential efficacy of piperine in protecting tissues from peroxidative damage. PMID- 11031727 TI - The influence of recombinant human granulocyte colony-stimulating factor on granulopoiesis in mice recovering from cyclophosphamide treatment. AB - Recombinant human granulocyte colony-stimulating factor (rhG-CSF) was evaluated for its effects on granulopoiesis recovery in mice pretreated with cyclophosphamide. The cytotoxic agent was administered on days 0 and 7. rhG-CSF therapy (injected from days 8 to 28) accelerated the recovery and maintained an increased level of peripheral blood neutrophils. Marrow granuloid precursors depression, due the second dose of cyclophosphamide, was prevented by rhG-CSF, but this hemopoietic growth factor was unable to increase these cells in a similar way to that observed in normal mice. This suggests a decreased granuloid marrow proliferation capacity in cyclophosphamide treated mice. In contrast, in the spleen, rhG-CSF highly increased granuloid precursors. However, the contribution of spleen to granuloid recovery was scarce. Mice receiving rhG-CSF after cyclophosphamide demonstrated a biphasic recovery pattern in bone marrow GM CFU population. A first rebound of GM-CFUs was followed by a nadir and then a second recovery where GM-CFU progenitor cells were significantly increased was observed. On the other hand, rhG-CSF therapy highly increased the spleen GM-CFU numbers at days 24 to 28. Although rhG-CSF increased splenic granulopoiesis, this result shows that the bulk of granulopoiesis recovery due to rhG-CSF therapy in cyclophosphamide pretreated mice occurred in the marrow. PMID- 11031728 TI - Enhancement of doxorubicin activity in multidrug-resistant cells by mefloquine. AB - We studied the effect of the antimalarial drug mefloquine on the resistance of K562 cells to doxorubicin. Mefloquine synergistically potentiated the cytotoxicity of doxorubicin for doxorubicin-resistant K562 cells (K562/DXR) at a concentration of 0.5-3 microM, but had hardly any synergistic effect in the parental cell line (K562) at the same concentration. Mefloquine was more potent than verapamil, a known modulator of multidrug-resistance. Since doxorubicin resistance in these cells is associated with the expression of high levels of P glycoprotein, we evaluated the effect of mefloquine and of P-glycoprotein activity in cytofluorographic efflux experiments with the fluorescent dye rhodamine 123. Our results indicate that mefloquine inhibits the P-glycoprotein pump-efflux activity in a dose-related manner. Moreover, mefloquine reduces the expression of the immunoreactive P-glycoprotein in K562/DXR cells as evaluated by cytofluorimetric assay. Taken together, the results indicate that mefloquine reverses the multidrug-resistance phenotype through direct interaction with P glycoprotein. PMID- 11031729 TI - Efficacy of fenfluramine and dexfenfluramine in the treatment of obesity: a meta analysis. AB - A meta-analysis was conducted to estimate the difference of weight loss among patients treated with placebo and with fenfluramine or dexfenfluramine after 1, 2, 3, 6, and 12 months of treatment. Placebo-controlled, double-blind, randomized clinical trials, whose results were presented as weight loss by the placebo group and the drug-treated patient group, were selected for the analysis. For the pooled estimations, the method of the weighted means by the inverse of the variance was used. The association between the difference of means and several predictive variables was studied by means of weighted linear regression. Patients treated with fenfluramine or dexfenfluramine achieved a higher weight loss than those receiving placebo in all the periods studied. The greatest efficacy was observed after 3 months of treatment. Beyond this time, there is a decline in the effectiveness. Based on the efficacy data, treatments longer than 3 months would not be justified. PMID- 11031730 TI - Cyclooxygenase isoenzymes and newer therapeutic potential for selective COX-2 inhibitors. AB - Cyclooxygenase (COX), also known as prostaglandin G/H synthase, is a membrane bound enzyme responsible for the oxidation of arachidonic acid to prostaglandins that was first identified over 20 years ago. In the past decade, however, more progress has been made in understanding the role of cyclooxygenase enzymes in various pathophysiological conditions. Two cyclooxygenase isoforms have been identified and are referred to as COX-1 and COX-2. COX-1 enzyme is constitutively expressed and regulates a number of housekeeping functions such as vascular hemostasis and gastroprotection, whereas COX-2 is inducible (i.e., sites of inflammation) by number of mediators such as growth factors, cytokines and endotoxins. Nonsteroidal antiinflammatory drugs (NSAIDs) produce their therapeutic effects through inhibition of COX, the enzyme that makes prostaglandins. Nonselective inhibition of COX isoenzyme leads to not only beneficial therapeutic effects but also a number of detrimental effects. Beneficial effects are due to inhibition of COX-2 and detrimental effects are due to inhibition of physiological COX-1. The present review discusses the biology as well as the role of these COX isoenzymes in various pathophysiological conditions. PMID- 11031731 TI - Nicotine transdermal systems: pharmaceutical and clinical aspects. AB - Smoking is creating a burden on the healthcare budgets all over the world. Management of nicotine dependence is difficult due to the so-called pleasurable effects experienced by smokers. Among the various treatment approaches, nicotine replacement therapy, where nicotine from cigarettes is replaced with nicotine from a relatively safer delivery vehicle, has been successful. Transdermal delivery of nicotine is an effective type of nicotine replacement therapy due to inherent pharmacokinetic advantages over other routes of delivery. Several nicotine transdermal delivery systems are available to suit the varying needs of smokers and have been found to be clinically patient friendly and well tolerated. This review focuses on the various clinical and pharmaceutical aspects of nicotine with special emphasis on transdermal delivery systems. PMID- 11031733 TI - Motivation and child maltreatment. Introduction PMID- 11031732 TI - Theophylline for the treatment of bronchial asthma: present status. AB - Current Japanese asthma guidelines recommend theophylline as an additional regimen to inhaled beta-agonists (on demand use) and inhaled corticosteroids (beclomethason dipropionate 200-1200 micrograms/day or fluticason propionate 200 800 micrograms/day) for patients whose asthma is not controlled completely. Similar recommendations are made in the International Consensus Report on Diagnosis and Treatment of Asthma. Recent advances in the understanding of its actions, especially regarding the antiinflammatory and immunomodulatory effects of low doses of theophylline, encourage its appropriate use in asthma treatment. Furthermore, asthma and chronic obstructive pulmonary disease (COPD) are considerably common diseases, and the economic benefits associated with the use of a relatively inexpensive, once or twice daily oral drug (intravenous infusion, if necessary) could be of importance. But, because of its narrow therapeutic range, dosage must be individualized in order to optimize the treatment based on the measurement of theophylline concentration in serum. Theophylline is an old medication with new forms. When used effectively and safely, it can have significant clinical benefits for patients. This paper intends to review the effectiveness of theophylline--historically and contemporarily. PMID- 11031734 TI - Motivation and mechanisms in the "cycle of violence". PMID- 11031735 TI - Social information processing and child physical abuse: theory and research. PMID- 11031736 TI - Developmental processes in maltreated children. PMID- 11031737 TI - Child abuse prevention: new directions and challenges. PMID- 11031738 TI - Balancing research and treatment in child maltreatment: the quest for good data and practical service. PMID- 11031739 TI - Understanding the dynamics of child maltreatment: child harm, family healing, and public policy (discussant's commentary). PMID- 11031740 TI - [Molecular mechanisms of drug transport]. AB - Drugs ingested into the body are transported through the plasma membrane several times. Although hydrophobic low molecules may penetrate the cell membrane according to simple diffusion, most drugs need carrier proteins named transporters for their trans-membrane transport. Drug transporters are classified into 5 groups by their difference in molecular structures, substrate specificities and transport mechanisms. The five groups of drug transports are as follows: (1) organic ion transporter superfamily, (2) ATP-dependent transporter superfamily, (3) peptide transporter family, (4) organic anion transporting polypeptide family originated from liver and (5) amino acid-polyamine-choline transporter superfamily. Among the 5 groups, there are seen several common characteristics such as structures having 12-transmembrane spanning domains, multispecificity in substrate recognition, different tissue distribution and functional regulation by phosphorylation. Multiple substrate specificities of drug transporters are tentatively understood by their evolution, in which at earlier stages these molecules might be essential to absorb endogenously necessary nutrients, and thereafter structural homologs like drugs might be recognized by these transporters. Several topics remain to be elucidated such as functional significance of drug transporters in pharmacokinetics including drug/drug interaction, polymorphism of drug transporter genes, and molecular mechanisms of substrate recognition by drug transporters. PMID- 11031741 TI - [Pharmacology of the glutamate receptor]. AB - The history of pharmacological examinations of glutamate receptor agonists such as kainic acid, quisqualic acid, acromelic acid, L-CCG-I, DCG-IV and L-F2CCG-I was described. Kainic acid is one of the most potent excitants in the mammalian central neurons, and its powerful excitatory actions gave rise to the excitotoxic concept that glutamate destroys neurons by excessive activation of excitatory receptors. Single systemic administration of acromelic acid, a kainate analog, caused behavioral and pathological effects quite different from those seen after systemic administration of kainate in the rat, demonstrating that the distribution of neuron damage caused by various excitatory amino acids is not always identical even if their receptors are in the same pharmacological category. 2-(Carboxycyclopropyl)glycine (CCG) is a conformationally restricted analogue of glutamate. CCG and its derivatives demonstrate unique neuropharmacological actions; for example, L-CCG-I and DCG-IV (a carboxylated derivative of L-CCG-I) relatively preferentially activate group II mGluRs. Prolonged infusion of very small amounts of DCG-IV showed a bell-shaped dose response relationship with regard to protection against kainate-induced neurotoxicity. Low concentrations of L-glutamate neither affected spinal reflexes nor the resting membrane potentials of motoneurons, but preferentially potentiated the depression of monosynaptic excitation caused by L-F2CCG-I. Following L-F2 CCG-I treatment, L-glutamate decreased the monosynaptic spinal reflexes in a concentration-dependent manner, indicating a 'priming' effect of L F2CCG-I. Thus, pharmacological actions of mGluR agonists are of great interest and remain to be clarified. PMID- 11031742 TI - [Nuclear receptor-mediated signaling pathway]. AB - Nuclear receptors for steroid/thyroid hormones, vitamins A and D, and fat-soluble ligands form a gene superfamily of ligand-inducible transaction factors, which plays important roles in a wide spectrum of biological events by regulating the expression of a set of target genes. DNA-bound nuclear receptors control transcription in a ligand-binding dependent way in cooperation with a multiprotein complex containing RNA polymerase II and a series of auxillary factors, TFIIA, B, D, E, F and H. During the process of ligand-induced transactivation by nuclear receptors, nuclear cofactors interacting with AF-1 and AF-2 seem to be involved. Several transcriptional co-activators and co-repressors forming coactivator complexes have been recently identified, and their function is discussed. PMID- 11031743 TI - [Repair and formation of cartilage and bone: are they possible?]. AB - Osteoporosis and osteoarthritis cause serious problems to the mobility, and therefore to the quality of life, of aged people. This is due to the fact that both diseases result from irreversible loss of quality or function of the tissues; i.e., bone and articular cartilage. Recent progress in molecular biology and developmental biology, however, have contributed a great deal to our understanding of the mechanisms regulating growth and differentiation of osteoblasts and chondrocytes, the cells responsible for producing bone and cartilage, respectively. In vivo studies using animals including humans also shed light on the mechanism underlying these biological events as well as diseases and provide ideas for possible interventions for treating these disease states. This review will deal with these new developments in the field and discuss possibility to develop new potential strategies for treating patients with those disorders. PMID- 11031744 TI - [A new approach to pharmacogenomics]. AB - The medicine in the 21st century will be so called "evidence based medicine" or "personalized medicine," based on the principle of "right drug to right patient." Pharmacogenomics covers the entire spectrum of genes that determines drug behavior and sensitivity, and we anticipate it will bring major impact on the healthcare system as well as the drug discovery process in the near future. Three waves of genomic impact are predicted to arise as follows: The first wave will hit on existing drugs and late-phase development candidates within the next 2-3 years, aiming to minimize the risks in clinical trials (adverse events, resistance, etc.). The wave will then affect the candidate selection process in the early pre-development stage, and finally the disease gene finding to target discovery process. The driving force will be technologies such as SNPs database, differential gene expression (DGE) analysis, proteomics, serial analysis of gene expression (SAGE) and bioinformatics. This new approach of genomic discovery (so called "integrated approach") requires knowledge on how to implement and integrate new valuable technologies from an early stage of the discovery process. The implication of SNPs, high throughput proteomics and application of structural genomics will be the key issues in the pharmacogenomics era. PMID- 11031745 TI - [Gene therapy--its present status and future direction]. AB - Remarkable progress in molecular biology has succeeded in developing gene therapy that is applicable in patients with a poor prognosis. At the Annual Meeting of Japanese Pharmacological Society this spring, the symposium entitled "Gene therapy: its present status and future direction" was held and this produced fruitful results. The symposium covered the following five topics, introduction to gene therapy, ischemic diseases, intractable heart failure, metastatic long cancer, ADA deficiency, which were presented by Drs. Y. Kaneda, Osaka Univ.; M. Aoki, Osaka Univ.; T. Kawada, Niigata Univ.; T. Fujiwara, Okayama Univ. and Y. Sakiyama, Hokkaido Univ., respectively. PMID- 11031746 TI - [Proposals for innovative pharmacology education from the perspective of several fields--in response to rapidly changing times]. AB - The Japanese Pharmacological Society has not given a symposium to discuss the topic of overall pharmacology education since 1994 (the 67th Annual Meeting). Thereafter, we have been witnessing great changes in the environment surrounding pharmacology, where scientific advances in life science have made boundaries between the various life sciences ambiguous and have brought the following questions before us: what is pharmacology and what should it be? Considering the situation, the Committee of Education of the Society organized a symposium on pharmacology education at the 73rd Annual Meeting in 2000, which will discuss current problems encountered in pharmacology education today and what pharmacology education should be hereafter. The symposists consisted of five people with backgrounds in basic medicine (biochemistry), clinical medicine (internal medicine), hospital pharmacy, pharmaceutical industry and administration (the Ministry of Health and Welfare). They unanimously emphasized that education of pharmacology should be reformed profoundly so as to contribute to a greater degree to professional medical services and new drug development as well as to basic science and meet the needs of the new era. We hope this symposium will trigger a further discussion on pharmacology education for the future. PMID- 11031748 TI - [Peripheral tissue microdialysis technique in unrestrained, conscious animals]. AB - The microdialysis procedure had been developed in the past 2 to 3 decades to determine levels of drug and endogenous compounds in several organs under physiological conditions. Advantages of microdialysis include: minimal stress on the experimental animals; studies may be done in unrestrained, conscious animals, and multiple determination can be made without concern for excess blood loss from small animals; and measurements can be made of drug and/or metabolites at multiple sites in the animal. By employing the microdialysis technique, I developed the method of multiple sampling for a long term period from an animal under the freely moving condition. In addition, I developed a novel microdialysis probe that was applied to several peripheral tissues and/or organs. In this paper, I will describe the fundamental procedure for peripheral tissues and/or organs such as the jugular vein and liver, using subcutaneous and ocular microdialysis sampling in unrestrained, conscious animals. PMID- 11031747 TI - [Pharmacological properties of nateglinide, rapid-onset/short-duration insulinotropic agent, in the treatment of type 2 diabetes]. AB - An early defect in Type 2 diabetes is the loss of acute insulin release after food intake, which causes prolonged elevation of postprandial glucose levels. Suppressing postprandial hyperglycemia is considered to be very important for preventing diabetic complications. Sulfonylureas are well-known insulin secretagogues and have been widely used in the treatment of Type 2 diabetes. These agents, however, do not appear to be able to ameliorate impairment of the first phase of insulin secretion and postprandial hyperglycemia. Nateglinide, which is a derivative of D-phenylalanine, is a non-sulfonylurea insulin secretagogue. Although the in vitro insulin-releasing effect of nateglinide is similar to that of sulfonylureas, its hypoglycemic effect is more rapid and short lasting. The in vivo unique pharmacodynamic profile of nateglinide is likely to result from its rapid absorption and elimination. This novel antidiabetic agent has made it possible to compensate for the impaired first phase insulin response and thus suppresses postprandial hyperglycemia. In clinical trials, nateglinide reduced prandial glucose excursion and improved early phase of insulin release dose-dependently after 12 weeks treatment. Nateglinide is a highly physiologic mealtime glucose regulator, which rapidly increases insulin secretion when taken before meals, mimicking early-phase insulin release lost in patients with Type 2 diabetes. PMID- 11031749 TI - [High-throughput techniques for analyzing SNPs and application of SNPs to pharmacogenomics]. AB - Progress in the human genome project can contribute to molecular diagnosis of various diseases and development of novel treatments. The next logical step for human genetics is the exploration and elucidation of genes involved in differential pharmacological response. An understanding of the role that genes have in pharmacological response is the cornerstone of personalized medicine. The pharmacogenomics approach is necessary and useful for this purpose. This article introduces these concepts to provide a context for the use of SNPs in pharmacogenomics and high-throughput techniques to analyze them. PMID- 11031750 TI - The flea genus Sigmactenus (Siphonaptera: Leptopsyllidae): three new taxa from Sulawesi, updated identification key, and distribution map for all known species and subspecies. AB - One new species and two new subspecies of fleas are described. These are S. sulawesiensis n. sp. from North and Central Sulawesi, S. alticola pilosus n. ssp. from Central Sulawesi, and S. alticola crassinavis n. ssp. from North Sulawesi. All three of these new taxa are ectoparasites of native, endemic murine rodents. Two of the new taxa, S. sulawesiensis and S. alticola crassinavis, coexist on the same mountain, Gunung Moajat, in North Sulawesi. The related S. alticola alticola, which becomes the nominate subspecies, parasitises the murine rodent Maxomys alticola in northern Borneo (Sabah) and it is hypothesized that Sigmactenus first colonized Sulawesi as an ectoparasite of ancestral Maxomys, or perhaps Rattus, as these murines dispersed from southeast Asia to Sulawesi; 15 endemic murine rodent species belonging to these two genera are known to currently inhabit Sulawesi. An identification key and distribution map are included for all known species and subspecies of Sigmactenus. In addition to the three new taxa and S. a. alticola, these include: S. celebensis from South Sulawesi, S. timorensis from Timor, S. toxopeusi from New Guinea, and S. werneri from the Philippines (Mindanao and Negros). PMID- 11031751 TI - [Cryptosporidium and wildlife: a risk for humans?]. AB - The present review underlines the knowledge of Cryptosporidium, especially its biodiversity and transmission. The presence of the parasite in different mammal host species is discussed with real, potential risk of transmission to humans. The potential role of insects in mechanical transmission of the parasite is evaluated by experimental protocols. The cost of cryptosporidiosis at health and economic levels are mentioned, which emphasises the importance of detection and identification of the parasite in the environment and in wild mammal species, using specific molecular tools. Potential measures to be accomplished in order to fight off cryptosporidiosis are also noted. PMID- 11031752 TI - A karyological study of the spirurid nematode Mastophorus muris (Nematoda: Spirocercidae). AB - The karyotype of Mastophorus muris (Gmelin, 1790) comprises four pairs of small autosomal chromosomes and two larger sex X chromosomes in females or one X chromosome in males (2n = 8 + XX/XO). All pairs of chromosomes possess rather uniform morphology without distinct primary or secondary constrictions. No heterochromatin bands were found by C-banding analysis. The absolute chromosome length ranges from 4.02 to 2.24 microns. The mean total length of the haploid complement is 14.34 microns. The course of gametogenesis represents a typical pattern common in the order Spirurida. The recently available karyotypes of spirurid nematodes have been reviewed. PMID- 11031753 TI - Spatial and temporal variations of the chromosomal inversion polymorphism of Anopheles funestus in Senegal. AB - The polymorphism of paracentric inversions of An. funestus polytene chromosomes was studied along a transect in Senegal in order to assess their variations at the spatial and temporal level. There was an increase in the degree of chromosomal polymorphism from the West to South-East. At the geographical level the variations in inversion frequencies were highly significant whatever the chromosomal arm considered. However, the variations in the chromosomal inversion frequencies did not change significantly over either seasons or years, except for inversion 3b in the village of Dielmo. Such geographical variability within a relatively limited area, associated to temporal stability, suggest a restricted gene flow between the populations studied, probably due to discontinuities in the An. funestus distribution and to its bioecology. PMID- 11031754 TI - Ultrastructural localisation of carbohydrates in four myxosporean parasites. AB - The existence and localisation of carbohydrates in four myxosporean parasites was investigated at transmission electron microscope, using lectin histochemistry techniques. The Myxosporea studied showed different lectin binding patterns. N acetyl-glucosamine or its polymers were predominant in the valves of Leptotheca sp. and were also present in Sphaerospora dicentrarchi and Polysporoplasma sparis ones. Mannose and/or glucose terminals were mainly detected in S. dicentrarchi and P. sparis valves. Polar capsule walls were intensively recognised by Con-A is S. dicentrarchi and with medium intensity in P. sparis and Leptotheca sp. The polar filament was scarcely labelled except in Leptotheca sp. with BS-I. The sporoplasms of the studied parasites were stained with all the lectins tested with each myxosporea, except those of Leptotheca sp. with BS-I. Some structures of the developmental stages of Zschokkella mugilis and Leptotheca sp. were stained with BS-I. The possible role of these carbohydrate terminals found in the studied Myxosporea is discussed. PMID- 11031755 TI - Fine structure of the development of Sarcocystis singaporensis in Python reticulatus from macrogamont to sporulated oocyst stage. AB - Three, 4-month old reticulated pythons (Python reticulatus) hatched from eggs laid by a newly caught female from Singapore Island, were fed on muscles of Sarcocystis singaporensis-infected Rattus rattus caught in Singapore. Snakes were sacrificed five, six and eight days later. The infected tissues were studied by transmission electron microscope. The present communication summarizes findings on macrogamont and oocyst stages. In the premature stages, rough endoplasmic reticulum consolidate into a large rectangular array; the electron-dense wall forming-like bodies reveal a laminar structure. Macrogamont parasitophorous vacuoles became filled with granular matrix and electron-dense strands, which later on consolidate into a coat around the fertilized zygote. The oocyst wall is constructed from several formed membranes combined with deposited substance. All development to the sporulated oocyst stage occurs in the mucosal epithelium. PMID- 11031756 TI - Sarcocystis stenodactylicolubris n. sp., a new sarcosporidian coccidium with a snake-gecko heteroxenous life cycle. AB - Oocysts/sporocysts of Sarcocystis sp. measuring 9.7 (9-10) x 7.6 (7-8) microns were found in the intestinal contents of the Dahl's whip snake Coluber najadum. Of wide spectrum of experimentally inoculated hosts, only species of the family Gekkonidae--Ptyodactylus guttatus and Stenodactylus grandiceps--were found to be susceptible intermediate hosts. Transparent, barely visible sarcocysts found in tail, limbs and tongue striated muscles of the geckoes were 175-200 microns x 35 50 microns in size at 78 DPI. Ultrastructurally, the primary cyst wall was characteristic by spine-like villar protrusions up to 800 nm long, 200-250 nm in diameter at their base, tapering to thinner apex. Protrusions appear typically lobular or irregular in the cross-sections. Back-transmission from P. guttatus to Coluber rogersi leaded to oocysts/sporocysts excretion since 38 days post infection. Based on sarcocyst morphology and experimental data, Sarcocystis stenodactylicolubris is apparently a new species. Based on obtained and already published results, Sarcosporidia parasitising colubrid snakes as definitive hosts are suggested to be family specific on the level of their intermediate host. PMID- 11031757 TI - Henneguya mbakaouensis sp. nov., Myxobolus nounensis sp. nov. and M. hydrocyni Kostoingue & Toguebaye, 1994, Myxosporea (Myxozoa) parasites of Centropomidae, Cichlidae and Characidae (Teleosts) of the Sanaga basin in Cameroon (Central Africa). AB - The study of 102 teleost freshwater fishes of Sanaga basin in Cameroon revealed the presence of three myxosporean species, among which two were new. Host fishes were of three families: Centropomidae, Cichlidae and Characidae. New species were identified as Henneguya mbakaouensis sp. nov., a gill parasite of Lates niloticus and Myxobolus nounensis sp. nov. found in the kidney and spleen of Sarotherodon galilaeus and Tilapia mariae. Myxobolus hydrocyni Kostoingue & Toguebaye, 1994, previously described in Chad, was also found in Cameroon; complementary informations were given on that parasite which seemed to be specific to its host. PMID- 11031758 TI - [Trichosomoides nasalis Biocca & Aurizi, 1961 and T. spratti n. sp. (Nematoda: Trichinelloidea), from the nasal cavity of murids]. AB - The nematode parasite of the nasal mucosa of rodents, Trichosomoides nasalis, is common in the murid Arvicanthis niloticus, in Senegal. The morphological study of this material allowed us to augment the original description, made with specimens recovered in Italy from Rattus norvegicus. This analysis shows that specimens from Rattus fuscipes in Australia, which had been identified to this species, represent a distinct species (smaller, with an average of five intrauterine males per female instead of one), which we name T. spratti n. sp. PMID- 11031759 TI - Efficacy of oxfendazole for the treatment of giardiosis in dogs. Experiments in dog breeding kennels. AB - Giardiosis is one of the most frequent parasites of dogs and cats. Since several years, the treatment is based on the use of metronidazole. A coproscopic study in four dog kennels was conducted to demonstrate, at a significant level, the efficacy of oxfendazole (Dolthene, Merial). At the posology of 11.3 mg/kg each day during three days (D1, D2 and D3), no dogs eliminated Giardia cysts and all dogs are clinically cured. The importance of hygienic measures is underlined. In kennels 1 and 2 where hygienic conditions were poor, dogs reexcreted cysts again after treatment. In kennels where the boxes were disinfected, no dogs, treated with 22.6 or 11.3 mg/kg, reexcreted Giardia cysts. PMID- 11031760 TI - Optimum conditions for growth in liquid medium of Oscillatoria formosa Bory used as the principal food in laboratory culture of intermediate hosts for schistosomosis and fasciolosis. AB - The rearing of snails, intermediate hosts of Schistosoma haematobium, S. intercalatum, S. bovis and Fasciola hepatica is the first step to maintain the life cycle of these parasites in laboratory in order to have biological material for the different studies, namely on the systematic biology and immunodiagnostic of schistosomosis and fasciolosis. According to the traditional method, the alga Oscillatoria formosa Bory (Cyanobacteria), principal food source for the snails, was cultivated in soil extract (Sampaio Xavier et al., 1968). However, it was sometimes very difficult to find the proper soil extract and the material was also contaminated by protozoa and fungi. In our work, using a new medium having as a base the Mineral Medium II (modified from Hughes et al., 1958) we found that O. formosa had a better growth response than in the soil extract medium. Snails fed on O. formosa reached three times the size of others at the same age, and they also reached sex maturity earlier, having more egg-masses per snail and, in addition, the rate of survival as well as the number of generations per year under laboratory conditions significantly increased. This culture was also easier to perform, and the axenic conditions easier to maintain. PMID- 11031761 TI - Cloning and expression of isocitrate lyase from human round worm Strongyloides stercoralis. AB - A full length cDNA (1463 bp) encoding isocitrate lyase (EC 4.1.3.1) of Strongyloides stercoralis is described. The nucleotide sequence of this insert identified a cDNA coding for the isocitrate lyase. The conceptually translated amino acid sequence of the open reading frame for S. stercoralis isocitrate lyase encodes a 450 amino acid residue protein with an apparent molecular weight of 50 kDa and a predicted pl of 6.39. The sequence is 69% A/T, reflecting a characteristic A/T codon bias of S. stercoralis. The amino acid sequence of S. stercoralis isocitrate lyase is compared with bifunctional glyoxylate cycle protein of Caenorhabditis elegans and isocitrate lyases from Chlamydomonas reinhardtii and Myxococcus xanthus. The full length cDNA of S. stercoralis was expressed in pRSET vector and bacteriophage T7 promoter based expression system. S. stercoralis lyase recombinant protein, purified via immobilized metal affinity chromatography, showed a molecular mass of 50 kDa on polyacrylamide gels. The role of isocitrate lyase in the glyoxylate cycle and energy metabolism of S. stercoralis is also discussed. PMID- 11031762 TI - Detection of acetylcholinesterase activity and gamma-aminobutyric acid binding sites in Dicrocoelium dendriticum. AB - In the present study we report the presence of acetylcholinesterase activity and gamma-aminobutyric acid binding sites in crude extracts of Dicrocoelium dendriticum. This indirectly demonstrates the presence of acetylcholine and GABA. The presence of these neurotransmitters could indicate the existence of two systems implicated in the neurotransmission of the Digenea. PMID- 11031763 TI - [Spatial distribution and bloodmeal preferences of Glossina palpalis palpalis in the forest focus of Zoukougbeu (Ivory Coast). Epidemiological consequences]. AB - In the sleeping sickness focus of Zoukougbeu (Cote d'Ivoire), in the cropping areas which are favourable for disease transmission, more than a quarter of the flies collected were found to have fed on domestic pigs. The sites where Glossina palpalis palpalis was caught fed on these animals were concordant with the sites where the patients were present. These results might indicate that in Zoukougbeu, but perhaps also in other sleeping sickness foci, the pig could play an active role in disease transmission, allowing the parasite to spread widely via the tsetse. PMID- 11031764 TI - Prophylactic and therapeutic application of antimicrobial agents in the oral cavity. AB - Antimicrobial substances are used for many prophylactic and therapeutic reasons in the oral cavity. They are used as mouthrinses, for the irrigation of specific oral locations, in form of gels, or varnishes. Different antimicrobial agents are available and some preparations are complex compositions of these compounds. Although it has been proven that several agents are suited to achieve a considerable reduction of oral microorganisms many questions concerning the clinical benefit are yet to be answered. This report summarises the evidence of the efficacy of oral antiseptics (especially chlorhexidine, pvp-iodine, and Listerine) depending on different indications, which is the basis for clinical and practical recommendations. PMID- 11031765 TI - L-ascorbic acid--clinical use, toxicity, properties, methods of determination and application in chemical analysis. PMID- 11031766 TI - [Potential antiallergics. 3. Synthesis and transformations of 1,4-dihydro-4-oxo (1)benzothieno(3,2-b)pyridine-2-carboxylic acid esters]. AB - The title compounds 2a, b are obtained by reaction of potassium 3-amino benzo[b]thiophene-2-carboxylate (1) with acetylenedicarboxylic acid esters in acetic acid. The substance 2a gives the carboxylic acid 2c by saponification, reaction with phosphoryl chloride affords the 4-chloropyridine 3. The carbinol 4, received by boranate reduction, was dehydrogenated with activated manganese dioxide to yield the carbaldehyde 5. Compound 2a reacts with methyliodide to give a separable mixture of the O- and N-alkylated products 6a and 7a, while by reaction of 2b with ethyliodide only the 4-ethoxypyridine 6b is formed. The carboxylic acids 6c, 7b are obtained by alkaline hydrolysis of the esters 6a, 7a. The carbinols 8, 9, formed by reduction of the esters 6a, 7a, are oxidized to give the carbaldehydes 10, 11. The 1H-tetrazoles 17, 18 are synthesized from the aldehydes 10, 11 via the aldoximes 13, 14 and the nitriles 15, 16. The aldehydes 10, 11 react with the beta-aminocrotonic acid esters 19 in acetic acid to yield the 1,4-dihydropyridines (DHP) 20, 21, which are dehydrogenated to form the pyridines (Py) 22, 23. The half-wave potentials E1/2 of the redox system DHP/Py is determined by difference pulse voltammetry using nifedipine as reference substance. PMID- 11031767 TI - [Synthesis and transformations of ethyl 1,4-dihydro-4-oxo(1)benzofuro(3,2 b)pyridine-3-carboxylic acid esters: new antibacterial agents]. AB - The title compound 7 was synthesized from potassium 3-amino-[1]benzofuran-2 carboxylate (1) by Gould-Jacobs-reaction. The pyridone 7 reacted with ethyl iodide by N- and O-alkylation to give 9 and 10, while methyl iodide only yielded the N-methylpyridone 11. The 4-chloropyridine 15 was obtained by heating 7 in phosphoryl chloride. Alkaline saponification of the esters 7, 9-11 and 15 afforded the carboxylic acids 8, 12-14 and 16. The carbaldehydes 19 and 22 were prepared from the ethylesters 10 and 15 by boranate reduction to the carbinoles 17 and 20 followed by dehydrogenation with activated manganese dioxide. The aldehyde 22 reacted with beta-aminocrotonic acid esters to yield the 1,4 dihydropyridines (DHP) 23. The pyridines 24 were formed by chemical or electrochemical dehydrogenation of the DHP 23. The tetrazole 27 was accessible from the aldehyde 22 via the aldoxime 25 and the nitrile 26. The pyridone 7 reacted with tosylisocyanate to yield the 4-tosylaminopyridine 28, which after alkylation to form 29 followed by detosylation to give 30 and subsequent alkaline hydrolysis produced the 4-aminonicotinic acid 31. The investigation The N ethylpyridone-3-carboxylic acid 12 showed antibacterial activity comparable to the reference substance nalidixic acid against Pseudomonas aeruginosa, Escherichia coli and Bacillus megaterium. PMID- 11031768 TI - In vitro recovery of triamcinolone acetonide in microdialysis. AB - The purpose of this study was to assess the factors affecting the calibration of the microdialysis probe for the in vitro recovery of triamcinolone acetonide (TA). Recoveries of TA were determined in microdialysis, retrodialysis, and no net flux methods. Experiments were performed at room temperature or 37 degrees C while the reservoir medium was either stirred or unstirred. The effect of the viscosity of the medium on the recovery was studied using methylcellulose gel spiked with TA. Recovery was also calculated by the no-net-flux method in Ringer's solution and in plasma. Stirring the medium increased the recovery of TA by 30%. The recovery was higher at 37 degrees C under stirred or unstirred conditions and was same in either direction of dialysis. Increasing viscosity of the reservoir medium decreased the recovery (55% in Ringer's solution to 14% in 20% methylcellulose gel). Recovery from spiked plasma under stirred conditions was only 15% and this shift which was also seen in no-net-flux method was accounted for by the protein binding. Binding of TA, determined by ultrafiltration, was 20% in 5% gel and 81% in plasma. The recovery determined by the no-net-flux method was similar to the retrodialysis result. Stirring, temperature, viscosity and protein binding in the reservoir medium affected the in vitro recovery of TA. PMID- 11031769 TI - Synthesis of iron-crosslinked hydroxamated alginic acid and its in vitro evaluation as a potential matrix material for oral sustained-release beads. AB - Hydroxamated alginic acid (HAA) was prepared. The generated semisynthetic polymer was employed in the formation of drug-loaded, iron(III)-crosslinked polymeric beads. Infrared spectroscopy was employed to prove the crosslinking. The degree of hydroxamate substitution was determined by acid-base back titration, and it was found to be ca. 20%. The produced beads were evaluated in vitro as orally administered sustained release drug delivery system. Theophylline, paracetamol, and chlorpheniramine maleate were used as model drugs. The generated beads proved to be successful in prolonging drug release, particularly in the case of theophylline. Iron leaching from the generated beads was minimal (< 0.5% of the entire complexed iron), and took place during passage through the simulated gastric fluid. PMID- 11031770 TI - Preparation and evaluation of bromocryptine mesylate-polydimethylsiloxane matrices. AB - The aims of the present study were to characterize the compatibility of silicone polymers with bromocryptine mesylate and excipients and to investigate the in vitro release characteristics of the drug from polydimethylsiloxane matrices. Silicon elastomers, MDX-4-4210 and A-2186, were chosen as polymer materials. Compatibility studies of polymers with drug and various liquid and solid excipients such as propylene glycol, polyethylene glycol, glycerol, sorbitan monolaurat, polysorbate 20, polysorbate 80, polyvinylpyrrolidone, citric acid, lactose, sodium chloride and low-molecular-weight gelatin were carried out. After the macroscopic examination of the excipient-polymer formulations, sorbitan monolaurat, propylene glycol, lactose, sodium chloride, citric acid and low molecular weight gelatin were chosen for investigation of the effect of these materials on drug release. Cylinder-shaped drug polymer matrices were prepared for the drug release studies. The best release profile was obtained with the formulation containing MDX-4-4210, 10% of propylene glycol and a kneading mixture of drug: low-molecular-weight gelatine in a ratio of 1:3. PMID- 11031771 TI - Influence of some DNA-alkylating drugs on thermal stability, acid and osmotic resistance of the membrane of whole human erythrocytes and their ghosts. AB - Human erythrocytes and their resealed ghosts were alkylated under identical conditions using three groups of alkylating antitumor agents: mustards, triazenes and chloroethyl nitrosoureas. Osmotic fragility, acid resistance and thermal stability of membranes were changed only in alkylated ghosts in proportion to the concentration of the alkylating agent. All the alkylating agents decreased acid resistance in ghosts. The clinically used drugs sarcolysine, dacarbazine and lomustine all decreased osmotic fragility and thermal stability of ghost membranes depending on their lipophilicity. DM-COOH did not decrease osmotic fragility and thermal stability of ghost membranes, while NEM increased thermal stability of membranes. The preliminary but not subsequent treatment of ghosts with DM-COOH fully abolished the alkylation-induced thermal labilization of ghost membrane proteins while NEM had a partial effect only. The present study gives direct evidence that alkylating agents, having a high therapeutic activity against malignant growth, bind covalently to proteins of cellular membranes. PMID- 11031772 TI - Influence of sexual steroids on cell functions of PMNL in the gingival sulcus. AB - Clinical experience confirms the influence of sexual steroids on the periodont under several clinical conditions. The mechanisms of the noticed effects are not all completely understood. In this paper, phagocytes from gingival crevice fluid of 39 patients with different forms of periodontitis and 18 healthy persons without periodontal disease were examined. Phagocytic activity was assessed in vitro. Simultaneously to phagocytic examination, 17 beta-estradiol or dienogest were added to the samples in different concentrations, in order to see whether a difference existed between phagocytosis in the presence or absence of sexual steroids. Phagocytosis was significantly reduced in patients with periodontitis. It was found that phagocytosis was raised significantly by 13% in the group with periodontal disease under the influence of 17 beta-estradiol. The administration of dienogest did not change the phagocytosis capacity significantly. In the healthy group, neither addition of 17 beta-estradiol nor addition of dienogest caused any difference. PMID- 11031773 TI - Antinociceptive activity of a hydroalcoholic extract obtained from aerial parts of Sebastiania schottiana (Euphorbiaceae). AB - This study analyzed the antinociceptive effects of a hydroalcoholic extract obtained from the aerial parts of Sebastiania schottiana, a Brazilian medicinal plant used to treat various painful diseases. For this purpose, the writhing test, capsaicin and formalin induced-pain in mice were used. The results showed that the hydroalcoholic extract exhibited considerable antinociception in all the models studied, being more potent than aspirin. PMID- 11031774 TI - Coumarin compounds in Ammi majus L. callus cultures. AB - Callus cultures of Ammi majus L. (bishop's weed), Apiaceae, were maintained on variants of Linsmaier-Skoog's (L-S) medium differing in the content of the phytohormones, alpha-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) (0.1-10.0 mg/l). The increments of callus tissue fresh weight showed considerable differences (1.4 bis 4.4-fold) during 4-week subcultures on the media tested. HPLC analysis revealed the presence of six compounds in the extracts of callus tissues, which are known metabolites in plants growing under natural conditions: the linear furanocoumarins psoralen, bergapten, xanthotoxin, isopimpinellin, imperatorin and their precursor umbelliferone. The total contents of coumarin compounds under examination showed marked differences dependent on the phytohormone concentrations in the medium. They ranged from 40.95 to 871.05 mg/100 g of dry weight. Imperatorin was the metabolite dominating among the furanocoumarins in calli from almost all tested variants of L-S medium (maximum content of 169.27 mg/100 g). Callus tissue cultured on one of the variants (0.1 mg/l NAA, 0.1 mg/l BAP) yielded in high content of xanthotoxin (145.33 mg/100 g). On the majority of media, umbelliferone was also accumulated at high quantities (maximum content of 536.29 mg/100 g). The medium containing 0.1 mg/l NAA and 0.1 mg/l BAP was the best for the accumulation of analysed coumarins. This medium favoured the formation of embryogenic callus. Xanthotoxin also dominated quantitatively among the furanocoumarins in the tested vegetative organs and fruits of the plant grown under natural conditions (leaves: 26.10 mg, roots: 5.55 mg, fruits: 3010.41 mg/100 g). Maximum contents of this metabolite in in vitro culture were many times higher than those found in vegetative plant organs but manifold lower than in fruits. On the other hand, maximum contents of imperatorin obtained in in vitro culture were many times higher in comparison with those detected in vegetative organs and fruits (leaves: 14.10 mg, roots: 3.30 mg, fruits: 94.70 mg/100 g of dry weight.). The A. majus L. callus culture, established in the course of the present experiments, can be considered a valuable model for studies of the biosynthesis of coumarin compounds, and a potential source of the psoralen 8-alkoxy derivatives imperatorin and xanthotoxin and their precursor umbelliferone. PMID- 11031775 TI - Phytochemical study and cytotoxic activity of alkaloids from Uvaria chamae P. Beauv. AB - Phytochemical study of leaves of Uvaria chamae resulted in the isolation for the first time for the genus Uvaria of benzylisoquinoline alkaloids (+)-armepavine (1) and racem. O,O-dimethylcoclaurine (2). The aporphines nornantenine (3), nantenine (4) and corydine (7) are new for the species. The alkaloids were found to express cytotoxic activity against L 929 transformed cells. The highest activity was shown by 1, 3, and 5. At a concentration corresponding to their IC50 against L929 cells, they were nontoxic against mouse thymocytes. PMID- 11031776 TI - Monodesmosidic saponins from Herniaria hirsuta. AB - Two new monodesmosidic saponins, herniaria saponins E and F, were isolated from the aerial parts of Herniaria hirsuta. On the basis of chemical and spectral evidence, their structures were established to be 2-O-acetyl medicagenic acid 28 O-beta-D-xylopyranosyl- (1-->4)-alpha-L-rhamnopyranosyl(1-->2)-[beta-D glucopyranosyl(1-->6)]-be ta- D-glucopyranoside (herniaria saponin E, 1) and medicagenic acid 28-O-beta-D-xylopyranosyl(1-->4)-alpha-L- rhamnopyranosyl(1-->2) [alpha-L-rhamnopyranosyl-(1-->4)-beta-D- glucopyranosyl-(1-->6)]-beta-D glucopyranoside (herniaria saponin F, compound 2). PMID- 11031777 TI - Two novel flavans from Cyperus conglomeratus. AB - Separation of the extract of the underground tubers of Cyperus conglomeratus Rottb. (family Cyperaceae) afforded, in addition to known compounds, two novel flavans, which were identified, by one and two dimensional NMR, MS and IR spectra, as 5-hydroxy-7,3',5'-trimethoxyflavan and 5,7-dihydroxy-3',5'-dimethoxy 6-prenylflavan. PMID- 11031778 TI - Diterpenoid alkaloids from the aerial parts of Aconitum anthora L. AB - Isolation and identification of six diterpenoid alkaloids, from Aconitum anthora L. are described. All proton detected 2D NMR techniques have been used for unambiguous 1H and 13C chemical shift assignments of guan-fu-base-Y (6). This is the first investigation of the diterpenoid alkaloids of this plant species. PMID- 11031779 TI - Separation of biflavonoids from Rheedia gardneriana using chitin-Fe complex as stationary phase. PMID- 11031780 TI - Resorcylidene aminoguanidine improves the pathologically reduced fluidity of erythrocyte membranes in diabetes mellitus. PMID- 11031781 TI - Effect of liquid extract from fresh Abutilon indicum leaves and Allium cepa bulbs on paracetamol and carbontetrachloride induced hepatotoxicity. PMID- 11031782 TI - Aframodial, a labdane diterpene showing selective in vitro antileukemic activity. PMID- 11031783 TI - Analgetic properties of glycosyl diclofenac derivatives. PMID- 11031784 TI - Prestimulation of monocytes by the cytokines GM-CSF or IL-2 enhances the antibody dependent cellular cytotoxicity of monoclonal antibody 17-1A. AB - Previously, we have shown that the cytokines IFN-alpha, IFN-gamma and IL-2 significantly enhance the antibody dependent cellular cytotoxicity (ADCC) exerted by the monoclonal antibody (mAb) 17-1A which recognizes the tumor associated antigen EpCAM. ADCC was assessed by a new flow cytometric cytotoxicity assay using the PKH2 labeled colorectal tumor cell line HT29 as target cells and peripheral blood mononuclear cells as effectors. Monocytes are assumed to be one of the major effectors for ADCC. However, isolated monocytes have a rather low ADCC capacity while addition of CD4+ lymphocytes optimizes ADCC. Since such an interaction between immune cells may act through cytokines we investigated whether a seven-day-prestimulation of monocytes by the cytokines M-CSF, GM-CSF, IFN-gamma, IFN-alpha and IL-2 enhances ADCC. Thereafter, we added for three days IL-2 and IFN-alpha with or without the mAb 17-1A for terminal activation of monocytes. Interestingly, GM-CSF which was ineffective in terminal activation, significantly enhanced ADCC of monocytes when it was used for prestimulation. Similar results were obtained with IL-2. IFN-gamma and M-CSF were also active but less than GM-CSF. Astonishingly, IFN-gamma and IFN-alpha prestimulation of monocytes suppressed the enhancement of ADCC exerted by GM-CSF and IL-2, respectively. Our experiments suggest that the timing of cytokine application is critical for the induction of optimal ADCC. Subcutaneous pretreatment with GM-CSF or IL-2 followed by the combination of IL-2/IFN-alpha/17-1A should be evaluated in a phase I clinical trial in patients with colorectal cancer. PMID- 11031785 TI - Fecal bacterial activity in symptomatic carbohydrate malabsorption: effect on the fecal short-chain fatty acid ratio. AB - BACKGROUND: It is still not clear why only some patients with carbohydrate malabsorption experience symptoms. In a previous study on healthy fructose malabsorbers an increased degradation of fructose in anaerobic fecal cultures from symptomatic malabsorbers was found, indicating increased bacterial activity. In the present study, the same investigation was repeated in patients with nonspecific abdominal complaints and fructose malabsorption. Moreover fecal short chain fatty acids (SCFA), products of colonic bacterial fermentation of carbohydrates were measured. PATIENTS AND METHODS: A standard quantity of fructose (500 mg) was added to anaerobic fecal cultures from 25 patients (nine men, 16 women; median of age 53 years, range 36-69 years). The fructose degradation rate was assessed using photometry, and interpreted as representing bacterial activity in the colon. In 14 of the patients, SCFA levels were also measured using chromatography on a capillary column. RESULTS: 10 of the 25 patients had a history of symptoms after ingesting fructose-containing foods, and also showed symptoms during the test; 6 patients had symptoms either in their history or during the test; and the remaining 9 were free of symptoms. There were no differences in the H2 increase. The fructose degradation rate was higher in symptomatic malabsorbers (255 mg vs. 217 mg), but the difference was not significant. However, there was a strong inverse correlation between this bacterial activity and the acetate level, with r = -0.822 (P = 0.000) and r = 0.868 (P = 0.000) in the rank correlation. The correlation for propionate was r = 0.479 (P = 0.083), and that for butyrate was r = 0.599 (P = 0.024). CONCLUSIONS: This study failed to confirm a significant correlation between fecal bacterial activity and the occurrence of symptoms in patients with fructose malabsorption. However, the interesting correlation with the SCFA raises questions regarding possible connections between colonic bacteria, carbohydrate malabsorption, and the beneficial effect of this pattern of SCFA in several colonic diseases. PMID- 11031786 TI - Comparative analysis of the Helicobacter pylori status in patients with gastric MALT-type lymphoma and their respective spouses. AB - Helicobacter pylori is of major importance for the development and progression of gastric MALT-type lymphoma. In case of localized low grade lymphoma successfully treated by Helicobacter pylori eradication reinfection by the bacterium may involve the risk of lymphoma reoccurrence. AIMS: To assess the frequency and type of Helicobacter pylori infection among patients with gastric MALT-type lymphoma and their respective spouses as potential sources of person to person spread of the infection. SUBJECTS: 35 patients with gastric MALT-type lymphoma and their cohabiting spouses. METHODS: We investigated serum IgG titers in response to the infection by Helicobacter pylori and to its virulence factors CagA and VacA by enzyme linked immunosorbent assay (ELISA) and by Western blot. RESULTS: Seropositivity of Helicobacter pylori, Cag A and Vac A was found to be 100/89/69% and 97/86/66% in patients and respective partners, respectively. The seroprevalence rates of the latter group by far exceed those of the German population. CONCLUSION: These data provide evidence for a high Helicobacter pylori infection rate in both patients with gastric MALT-type lymphoma and their respective spouses. Considering the latter as a potential source of reinfection with the risk of lymphoma reoccurrence for the successfully treated patient with gastric MALT-type lymphoma careful follow-up seems highly reasonable to decide on the necessity of a future test-and-treat strategy of this population. PMID- 11031788 TI - Gastric outlet obstruction and pulmonary infiltrate in a patient with Crohn's disease: successful treatment by Billroth-II-resection. AB - We present a 28-year-old women with a 3 yr history of duodenal ulcers. Following four treatment attempts to eradicate helicobacter pylori she was admitted because of gastric outlet obstruction and a weight loss of 20 kg within the last two years. Endoscopy and x-ray showed a circular inflammatory stenosis of the proximal duodenum extending over 8 cm. Additionally, chest x-ray showed a circumscript infiltrate in the third segment of the right lung. Mycobacterial infection could be excluded. Ileocolonoscopy and small intestinal follow-through beyond the duodenum were unremarkable, and Zollinger-Ellison-syndrome was ruled out. Bronchopulmonary histology showed intramucosal epithelioid-cell granulomas and bronchiolitis obliterans. Because the patient did not improve under conservative therapy a Billroth-II-resection was carried out. Histologically the resected specimen showed Crohn-like lesions. Postoperatively, severe peripheral arthritis was treated by steroids over 6 weeks. At follow-up the patient regained 20 kg and was free of symptoms without any medication. The pulmonary infiltrate had subsided almost completely. In summary, this extremely rare coincidence of isolated stenosing duodenal Crohn's disease and pulmonary involvement was successfully treated by Billroth-II-resection. This course of disease is compatible with the hypothesis that Crohn's disease may be maintained by antigens derived from ingested food. PMID- 11031787 TI - [Gastrointestinal hemorrhage needing blood transfusion as the first manifestation of small bowel carcinoid tumor]. AB - Carcinoid tumors arise from enterochromaffin or enterochromaffin-like cells that are present in the gastrointestinal tract, ovaries, and lungs. Over 90% of carcinoids originate in the gastrointestinal tract with the most common sites in order of frequency being the appendix, terminal ileum, rectum, and the remainder of the colon. Gastroduodenal and pancreatic carcinoids are infrequent. Carcinoid syndrome is associated with small intestine carcinoids in about 40%. Common symptoms include intermittent intestinal obstruction with crampy abdominal pain and vomiting, and weight loss. Upper gastrointestinal bleeding with melaena or hematochezia is a relatively rare early symptom of patients with small intestine carcinoid tumors. We report on a 69-year-old man, treated with acenocoumarol for previous thromboembolic complications of hereditary protein S deficiency. He was admitted to hospital because of an acute episode of hematochezia followed by melaena. Endoscopic evaluation of esophagus, stomach, duodenum and colonoscopy revealed no apparent source of bleeding. Selective angiographic evaluation of mesenterial arteries showed pathologic vasculature approximately in mid jejunum. Laparotomy revealed bleeding from a small submucosal malignant carcinoid tumor in small intestine and multiple large metastases within mesenteric tissue. Segmental resection of small intestine and exstirpation of the metastatic masses was performed. Postoperative period was uneventful. Cytotoxic chemotherapy in this adjuvant setting has not been recommended. Small intestinal carcinoid tumor has to be considered as a rare cause of gastrointestinal bleeding with melaena or hematochezia. Nevertheless, bleeding is a relatively rare early symptom of patients with small intestine carcinoid tumor. PMID- 11031790 TI - Modulation of gastrointestinal inflammation by chimeric proteins in experimental models. AB - Conventional drug therapy in patients with chronic gastrointestinal inflammation is clinically effective in the majority of patients. However, in a relevant group of patients with highly active disease refractory to conventional drugs and for patients with severe side effects new therapeutic strategies are necessary. An advanced understanding of the immune mechanisms underlying chronic diseases resulted in the possibility to use chimeric proteins, in which the variable domains of an immunoglobulin are replaced by extracellular domains of cell surface molecules or cytokines for specific immunomodulation. The immunomodulating effects of chimeric proteins such as CTLA-4-IgG, interleukin-10 IgG, IL-2-IgG or tumour necrosis factor (TNF)-receptor IgG have been proven beneficial in a variety of in vitro and in vivo models of chronic gastrointestinal inflammation and autoimmune diseases. It thus seems likely that genetically engineered fusion proteins targeting specific elements of the immune response may become an essential element in new clinical treatment protocols. PMID- 11031789 TI - [Malacoplakia-like reaction in association with colorectal adenocarcinoma]. AB - 17 cases of malacoplakia in association with an adenocarcinoma of the colon or rectum are known. In this report we describe a new case of this rare combination. An 83-year-old comatose female patient was admitted in an emergency situation because of an incipient ileus. A carcinoma had been known for months, but the patient had previously rejected any diagnostic or therapeutic measures. A tumor (6 cm in diameter) in the upper rectum was removed. This tumor had caused an obstruction of the lumen and reached the serosa. Histologically, an adenocarcinoma with suppurations in the stroma was staged pT3, G2, pN0 (clinically pT4 because of the perforated colonic wall). PAS- and von-Kossa positive Michaelis-Gutmann-bodies were found in macrophages. Ultrastructurally, they turned out to be circular shifted early stage calcifications. Any malacoplakia-like lesion in suspicious areas of the colonic wall should be followed up by further diagnostic measures, since it may indicate the presence of an adenocarcinoma. PMID- 11031791 TI - [Transforming growth factor beta as a protective factor in autoimmune pancreatitis]. PMID- 11031792 TI - [TIPS: value in the treatment of therapy refractory ascites]. PMID- 11031793 TI - [34th Scientific meeting of the German-speaking Association of Mycology. Berlin, Germany, September 14-16, 2000. Abstracts]. PMID- 11031794 TI - [XXX National Congress of the Spanish Society of Nephrology. 8-11 October 2000. Abstracts]. PMID- 11031795 TI - 9th International Conference on Behcet's Disease. May 27-29, 2000, Seoul, Korea. Abstracts. PMID- 11031796 TI - Essential medical facts for mental health practitioners. AB - New information about the life cycle of HIV, new HIV-specific laboratory tests, and newer antiretroviral medications have transformed the management of HIV illness. Knowledge about these changes will help mental health providers better understand the latest medical issues affecting their HIV-infected patients, which will assist them in providing better care. PMID- 11031797 TI - Psychosocial issues in antiretroviral treatment. AB - The advent of effective treatments for HIV has begun a new era in the worldwide HIV epidemic. Many new political, social, economic, medical, and psychological issues arise in the struggle to contain this epidemic. Mental health providers must understand the context in which people with HIV find themselves making decisions about their health care and the future directions of their lives. PMID- 11031798 TI - Diagnosis and treatment of HIV-associated neurocognitive disorders. AB - Although significant strides have been made in recent years in treating HIV disease with new antiretroviral medications, the management of neurocognitive disorders continues to remain a challenge. This chapter provides an overview of the current epidemiology, neuropathogenesis, clinical features, diagnosis, and treatment of the central nervous system complications of HIV infection. PMID- 11031799 TI - Psychotherapeutic strategies for coping with HIV infection. AB - Psychotherapy promotes adaptation to the medical, neuropsychiatric, and psychosocial challenges faced by people with HIV infection. This chapter describes a range of individual and group psychotherapy techniques with demonstrated efficacy in this population. PMID- 11031800 TI - Some further thoughts on adjustment, coping, and bereavement. AB - Adjusting to and coping with HIV-AIDS is a difficult task. This chapter describes some of the issues surrounding being diagnosed HIV positive, living with HIV, and progressing to AIDS. It also explores bereavement reactions that relatives and friends can experience in dealing with illness and death due to HIV. PMID- 11031801 TI - Mood and anxiety disorders. AB - This chapter describes two of the most frequent psychiatric conditions occurring in people with HIV and AIDS, namely, anxiety disorders and depression, and describes the commonly occurring features, clinical correlates, and treatment considerations for these disorders. PMID- 11031802 TI - Substance abuse and HIV infection. AB - This chapter reviews the associations between substance use, comorbid psychiatric disorders, and HIV risk behaviors; the prevalence of substance use disorders among HIV-positive individuals in treatment settings; the medical, psychiatric, and substance abuse treatment of individuals with substance use disorders and HIV infection; and finally, HIV risk reduction among substance abusers. PMID- 11031803 TI - Severe mental illness and HIV-AIDS. AB - This chapter reviews the high rates of HIV infection and the risky sex and drug use practices of psychiatric patients, examines the role in HIV risk of psychiatric symptoms and of the contexts in which patients live, provides guidelines for taking sexually transmitted disease and risk histories from patients, presents recommendations for making HIV-testing decisions with patients, and describes the range of HIV-related services available to psychiatric outpatients. PMID- 11031804 TI - Preventing HIV infection through sexual-behavior change. AB - Research on HIV prevention during the past decade has identified numerous models, at levels ranging from individual interventions to public policies capable of affecting an entire population. None of these approaches should be considered sufficient in itself because, put simply, behavior is difficult to modify. This truism is especially applicable when the behavior is multiply determined and serves many needs, as is the case with sexual behavior. Therefore each of these approaches should be seen as one part of a comprehensive HIV prevention program. Such efforts need to target individuals, small groups, communities, schools, and social policies. Adoption of theoretically guided and empirically validated approaches promise the most success. PMID- 11031805 TI - Adherence to treatment. AB - Since the advent of highly active antiretroviral therapy, the issue of strict adherence has become increasingly important. This chapter examines how the mental health provider can employ a multimodal approach to promoting patient adherence, which increases the chances of success. PMID- 11031806 TI - Legal and ethical issues relevant to HIV-positive psychiatric patients. AB - Three major legal and ethical issues central to the AIDS epidemic--capacity to consent to testing, degree of dangerousness, and duty to warn third parties--are examined, as they influence the management of psychiatric patients. PMID- 11031807 TI - Provider issues in HIV care. AB - This chapter examines the impact of HIV infection and its associated biopsychosocial complications on provider-patient relationships and addresses the implications for health care teams and organizations. Several interventions are suggested that can relieve the complex stresses that health care providers are currently experiencing as they care for HIV and AIDS patients. PMID- 11031808 TI - An integrated program for comprehensive HIV care. AB - HIV-AIDS medical care systems that do not have full integration of mental health, alcohol, and substance use services--including the key components of prophylaxis, medication adherence, and outpatient symptom management--risk less-than-optimal outcomes. This chapter provides a model for integrated treatment. PMID- 11031809 TI - A program for families and children affected by HIV. AB - HIV affects not only the individual who contracts the disease but also members of that person's family. The Family Center specializes in providing services to help parents and children deal effectively with the demands of the illness and to ensure that the children are cared for after the parent's death. PMID- 11031810 TI - A practitioner's view from the front lines. AB - One practitioner's experience mirrors the changing face of the AIDS epidemic. PMID- 11031811 TI - [Recently advanced research on allergic conjunctivitis and atopic dermatitis]. PMID- 11031812 TI - [Subretinal surgery for choroidal neovascularization]. AB - BACKGROUND: Ten years have passed since subretinal surgery for choroidal neovascularization (CNV) was developed. Many reports regarding the visual outcome after subretinal surgery have been published, which have revealed the role of the subretinal surgery, including long-term visual outcome and surgical complications. METHOD: Reports associated with subretinal surgery published in the past decade were used to clarify the indication for surgery, visual outcome, and surgical complications. RESULTS: The visual prognosis for patients with surgically excited CNV is variable, depending on the underlying disease. Patients with widespread defects of Bruch's membrane and retinal pigment epithelium (RPE), such as age-related macular degeneration, are probably not good candidates for surgery from the standpoint of recovering good visual acuity. On the other hand, patients with focal abnormalities in Bruch's membrane and the RPE, such as presumed ocular histoplasmosis or idiopathic CNV, may be good candidates for surgery. However, it has not been proved that subretinal surgery is superior to photocoagulation or observation. CONCLUSIONS: For the time being, we have to select patients for subretinal surgery on the basis of published reports. The Submacular Surgery Trial now being held in the United States will evaluate the role of subretinal surgery in the management of patients with choroidal neovascularization. PMID- 11031813 TI - [Sialic acid in normal human tears]. AB - PURPOSE: We measured the concentration of sialic acid, the terminal component of mucin, in normal human tears. SUBJECTS AND METHODS: Tear samples were collected from 31 healthy volunteers(50.7 years old on average) using micropipett after instillation of saline. We investigated the correlation of concentration between glycoprotein and sialic acid, the difference between these in the right eye and left eye, and the reproducibility of the sampling procedure. RESULTS: There was significant correlation of concentration between glycoprotein and sialic acid(right eye: r = 0.989, p < 0.0001; left eye: r = 0.988, p < 0.0001). There was no significant difference between these in the right and left eye. Also, the reproducibility was considered acceptable in three measurements of sialic acid with the present procedure. The concentration of sialic acid in normal diluted tears was 37.1 micrograms/ml on average. CONCLUSION: Our data indicates that concentration of sialic acid reflects that of glycoprotein and this method of measurement is applicable to the analysis of disorders with mucin deficiency. PMID- 11031814 TI - [Influence of cataract on scanning laser polarimetry]. AB - PURPOSE: To evaluate the influence of cataract on scanning laser polarimetry. METHODS: Nerve fiber layer thickness (NFLT) around the optic nerve head was measured before and after surgery in 43 eyes of 43 patients with cataract. Phacoemulsification-aspiration and intraocular lens implantation was performed in all eyes. RESULTS: The mean NFLT increased 13.41% (p < 0.0001) postoperatively. Positive correlation(rs = 0.56, p = 0.001) was observed between NFLT measurements and cataract degree, using Lens Opacities Classification System III. Significant positive correlation(rs = 0.493, p = 0.004) was observed between NFLT measurements and posterior subcapsular cataract degree. CONCLUSION: Scanning laser polarimetry may underestimate NFLT in individuals with dense cataract. PMID- 11031815 TI - [Pattern visual evoked potentials visual acuity--the evaluation of visual acuity of less than 0.1]. AB - PURPOSE: We previously developed a new method for estimating objective visual acuity by means of pattern visual evoked potentials (PVEP). In this study, this method was applied to the evaluation of visual acuity of 0.05 and 0.01. METHODS: Ten subjects with visual acuity of 0.05 and ten normal subjects with visual acuity of 0.01 were examined. The visual acuity of the subjects was decreased step by step by means of plus spherical lenses of increasing power. In the PVEP study, the stimulus consisted of black and white checkerboards with a visual angle of 8 degrees (central stimulus) and a global field with a 5 degrees central circular scotoma (peripheral stimulus), contrast level of 15%, and a frequency of 0.7 Hz. One hundred PVEP responses were averaged per session. We reported that there was a linear relation between log subjective visual acuity of 0.1-1.0 and log check size of the patterns with PVEP. In reference to that, the check sizes of the patterns were estimated at 66' with visual acuity of 0.05, and 107' with visual acuity of 0.01. Each visual acuity was then used with other check sizes of three patterns. RESULTS: With central stimulus the P100 components of O1, O2, and Oz were recorded with 66' and 82' patterns at visual acuity of 0.05(central stimulus). The P100 components of O1, O2, and Oz were 107', 137' and 161' at visual acuity of 0.01(central stimulus). There was no P100 component at visual acuity of 0.05 and 0.01(peripheral stimulus). CONCLUSION: This method of PVEP is a useful tool as an objective estimation of visual acuity less than 0.1 and is presumed to stimulate preferentially the X retinal ganglion cell to parvocellular pathway. PMID- 11031816 TI - [Immunological and clinical features of vernal keratoconjunctivitis between persons with and without systemic atopic dermatitis]. AB - PURPOSE: The aim of this study was to evaluate whether there exists immunological or clinical difference between patients with vernal keratoconjunctivitis(VKC), which is characterized by conjunctival proliferative changes belonging to the serious spectrum of allergic conjunctival diseases, complicated by atopic dermatitis(AD) and those without AD. MATERIALS AND METHODS: The study population consisted of 52 patients with VKC (33 patients with AD, 19 without AD). Clinical severity was determined on the basis of clinical scores of conjunctival, corneal, and limbal lesions. The serum level of total IgE, the proportion of eosinophils in peripheral blood, antigen specific IgE antibodies (radioallergo sorbent test, RAST), and tear levels of interleukin (IL)-4 and IL-5 were measured. RESULTS: The serum level of total IgE and the number of positive RAST antigens were significantly higher in VKC patients with AD than in those without AD. No significant difference was observed in the proportion of peripheral blood eosinophils, distribution of clinical types of VKC, or clinical severity between VKC patients with and without AD. Although tear IL-4 level in VKC patients with AD was significantly higher than in those without AD, no difference was observed in tear level of IL-5 between these groups. CONCLUSIONS: These results indicate that clinical features are similar or identical in VKC regardless of the presence of AD. However, it is suggested that there may be a different immunological background between VKC patients with AD and those without AD. PMID- 11031818 TI - [Geriatric home health care]. PMID- 11031817 TI - [Siblings with age-related macular degeneration in a pedigree]. AB - BACKGROUND: The pathogenesis of age-related macular degeneration remains unknown. Environmental and genetic factors have been shown to be associated with in this disease. We report siblings with age-related macular degeneration. CASES: The proband (the third of 6 siblings) was found to have retinal pigment epithelial abnormalities and drusen of the right macula, and serous retinal detachment, drusen, and choroidal neovascularization of the left macula. Examination of the eldest siblings showed choroidal neovascularization, subretinal hemorrhage, and serous retinal detachment of the right macula. The left eye of the proband and the right eye of the eldest siblings were treated with laser photocoagulation. The fourth sibling was examined, and findings included retinal pigment epithelial abnormalities of both maculas and drusen of the right eye. The sixth sibling had drusen of the left macula. CONCLUSION: We report a family in which two siblings had age-related macular degeneration and two other siblings had abnormalities of the maculas. PMID- 11031819 TI - [The biology of menopause]. PMID- 11031820 TI - [Long-term care insurance in Germany--similarity and problems in Japan and Germany]. PMID- 11031821 TI - [Elder care systems and practices in the U.S.A]. PMID- 11031822 TI - [Overview and future forecast of public long-term care insurance--from a critical point of view]. PMID- 11031823 TI - [Health services facility for the elderly and long-term care insurance in Japan]. PMID- 11031824 TI - [Differences between the first and second appraisals in a new social insurance system covering long-term care service--in relation to the presence of dementia]. PMID- 11031825 TI - [The long-term care insurance and medical care for the elderly--the perspectives for the insurance's implementation]. PMID- 11031827 TI - [Neuropsychological comparison between corticobasal degeneration and progressive supranuclear palsy]. AB - We conducted a neuropsychological comparison among cases with corticobasal degeneration (CBD; n = 8), those with progressive supranuclear palsy (PSP; n = 5) and healthy control subjects (n = 12) using an extensive neuropsychological battery assessing memory and executive functions. There were no significant differences among three groups for age, education, scores on the Mini-Mental State Examination and Zung's self-rating depression scale. Both patient groups showed retrieval impairment without recognition difficulties, and a dysexecutive syndrome. Along with those similarities, we observed some differences between CBD and PSP patients. Memory impairments in CBD patients were more marked than PSP patients in Rey's complex figure test, while they were less prominent in Rey's auditory verbal learning test. Perseverative errors of Nelson in Wisconsin card sorting test (Keio version) were more marked in CBD patients than in PSP patients. These two diseases showed memory and executive dysfunctions probably due to subcortico-frontal dysfunction. Some neuropsychological differences may help to distinguish CBD clinically from PSP. PMID- 11031826 TI - [Plasma cardiac natriuretic peptide as a biological marker of recurrence of atrial fibrillation in elderly people]. AB - We designed this study to evaluate the relationship between plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) levels and recurrence of atrial fibrillation (AF) after direct current cardioversion (DC) and the differences with aging. Fifty patients with mild congestive heart failure (CHF) undergoing elective DC of AF were included in this study (New York Heart Association (NYHA) functional class II: n = 42, III = 8). Patients who failed to show restoration of sinus rhythm or those with mitral valve stenosis were excluded. Before successful DC, we measured plasma levels of ANP and BNP and evaluated left atrial dimension (LAD), left ventricular end-diastolic dimension (LVDd), and left ventricular ejection fraction (EF) by echocardiography. Twenty one patients had recurrence of AF within 2 months after DC (average 9.05 days). We followed up the other 29 patients for 580.5 days. By Cox stepwise multivariate analysis, history of AF (p = 0.007), low plasma levels of ANP (p = 0.003), and high plasma levels of BNP (p = 0.0003) were found to be independent predictors of recurrent AF. High plasma BNP levels indicating ventricular dysfunction and low plasma ANP levels may be due to atrial histological change such as fibrosis. In these patients, plasma ratios of ANP and BNP (ANP/BNP) less than 0.43 were predictive factors for AF recurrence (sensitivity 70%, specificity 62%), especially in patients who were older than 70 years (sensitivity 100%, specificity 80%). Relatively low plasma ANP level compared to BNP is an independent risk factor of AF recurrence in patients with CHF, especially in elderly patients, suggesting that plasma cardiac natriuretic peptides are important biochemical markers of AF recurrence in elderly patients with CHF. PMID- 11031828 TI - [Behavioral competence among community dwelling older people with disability in basic activities of daily living]. AB - This study observed the status of independence in behavioral competence among older people who have any disability in basic activities of daily living (BADL) living in a rural community in Japan. Study participants (N = 76) who were regarded as bedridden were surveyed by means of a questionnaire in July to August 1996. The independence variables were age, sex, BADL status, hearing impairment, visual impairment, history of stroke, and cognitive impairment. The dependent variable was each item of the Tokyo Metropolitan Institute of Gerontology (TMIG) Index of Competence, which is a multidimensional 13-item index of behavioral competence. Percentages of subjects who were independent in each item of the TMIG Index of Competence varied from 1% to 36%. Multiple logistic regression analyses revealed that BADL status was independently associated with independence in using a telephone, being interested in news stories or programs dealing with health, being called on for advice, and initiating conversations with young people, after adjustment for age, sex, hearing impairment, visual impairment, history of stroke, and cognitive impairment. These findings suggest that programs for preventing decline in behavioral competence of older people with BADL disability might be important as well as physical therapy for them. PMID- 11031829 TI - [Influential factors on health-related quality of life after cerebral vascular disease]. AB - A mail survey was conducted to elucidate the influential factors on heath-related quality of life (HRQOL) after cerebral vascular disease. Questionnaires for clinicians and their patients were mailed to 2,587 hospitals with more than 100 beds, which have at least one of the following departments: neurosurgery, neurology, psychiatry or rehabilitation. Each mailing contained a request to the clinician and questionnaires for 5 cases. 378 effective questionnaires could be collected, meaning the collection rate was 2.9%. The questions for the physicians concerned diagnosis (cerebral infarction or hemorrhage), duration of illness, activities of daily living(ADL), manifestation of paralysis and psychiatric symptoms and so forth. The questionnaire for the patients was composed of items from the EuroQol clinical version (EuroQol). Geriatric Depression Scale short form (GDS) and inquiries concerning family living with the patients, their housekeeping and so on. A visual analogue scale (VAS) concerning health state of the EuroQol was used as a measure of HRQOL. Coefficients of determination between VAS and other inquiries were calculated by regression analysis or ANOVA, revealing that "anxiety/depression", "GDS" and 16 other items were statistically significant on VAS (p < 0.05). General linear model (GLM) analysis using VAS as a criterion variable and these 18 items as predictor variables showed that "sleep disturbance" and GDS score were most influential on VAS according to the F value of the type 3 sum of squares. "Health state today compared to that during the past one year", "shopping as housekeeping", "ADL" and "pain/discomfort" also have some influence on VAS. In conclusion, sleep disturbance and depression had the most deleterious effect on HRQOL. PMID- 11031830 TI - [A very elderly autopsy case of cecal cancer with pulmonary lymphangitis carcinomatosa]. AB - A 91-year-old man was admitted with colliquative diarrhea, anorexia and weight loss. He had a history of healed tuberculosis, hypertension and atherosclerotic abdominal aortic aneurysms. On admission, shortness of breath without cough, exertional dyspnea, and ascites were also noticed. His chest X-ray and CT showed almost normal findings in the lung fields except for calcified old pleurisy. Since laboratory tests revealed thrombocytopenia, low fibrinogen, and increased CA19-9. DIC induced by an unknown cancer was diagnosed. He died on the eighth day due to progressive respiratory failure which did not respond to oxygen therapy. Autopsy revealed that he had a poorly differentiated adenocarcinoma in the cecum complicated with pulmonary lymphangitis carcinomatosa. Lymphangitis should be considered in the case of unexplained progressive respiratory failure developing in patient with cancer, even in the absence of X-ray findings. PMID- 11031831 TI - [A case of percutaneous and transpapillary placements of expandable metallic stents in a patient with cholangiocarcinoma at the hilum of the liver]. AB - A 76-year-old woman was admitted with obstructive jaundice. US and MR cholangiopancreatography (MRCP) revealed an inoperative cholangiocarcinoma, 3 cm in diameter at the hilum of the liver, the obstruction of the hepatic duct bifurcation and the separation of bilateral hepatic bile ducts. Percutaneous transhepatic biliary drainage (PTBD) was performed from bilateral hepatic bile ducts. The right PTBD tube was spontaneously extubated. We could not succeed in performing internal biliary drainage across the hilar malignant stricture from a left hepatic bile duct, because of bad angulation. Transpapillary insertion into the common bile duct (CBD) was extremely difficult due to the collapse of the CBD. Endoscopic sphincterotomy (EST) after precutting method was performed. Although we performed the ballooned dilatation of malignant stricture and the insertion of a self-expandable metallic stent (EMS) into a right hepatic bile duct transpapillary. After dilatation of the hilar malignant stricture by the initial EMS, we inserted a guidewire into the CBD through the wire mesh of a stent from the left PTBD tube. We could insert the second EMS from a left hepatic bile duct to the CBD transhepatically, using a dilator and a dilating balloon. Finally, we performed the ballooned dilatation from bilateral hepatic bile ducts to the CBD transpapillary. She was discharged after bilateral internal biliary drainages, successfully. PMID- 11031832 TI - [Comparative analysis of quantifying methods in FDG-PET scans of the brain]. AB - The comparative analysis of three tracer kinetic methods most frequently applied for the quantization of the results of FDG-PET-brain scans was performed. The data of measurements on five healthy patients were evaluated by the most general method developed by Phelps, the Patlak-procedure and the SUV- (standard uptake value) method. It was demonstrated by the aid of correlation analysis that the applicability of the results of the more simple methods to estimate glucose metabolic rate (GMR) as calculated by the Phelps-method depended on the kind of the selected region of the brain. It was shown that the most considerable distortion occurred in the case of the same anatomical regions of the brain with both simplified methods. These regions were located either in the white matter or in the vicinity of larger size blood vessels or they were elements of the base of the skull [gyrus rectus (l. u.), pons, capsula interna (l. u.), cerebellum (l. u.), corpus callosum]. The distorted estimation is explained by the fact that the simpler models neglect dephosphorylation of the FDG-6P, and they also disregard the contribution of the intravascular activity to the tissue radioactivity as determined by the relatively low resolution PET measurement. The correlation coefficient between the GMR as calculated by the Phelps-method and glucose consumption data by the investigated simpler methods had very low values for regions located in the white matter, eventual close to blood vessels or being elements of the base of the skull. PMID- 11031833 TI - [Low birth weight, adrenal and sex hormones and their correlation with carbohydrate metabolism and cardiovascular physiology, investigated in young adulthood] . AB - It is known that the prevalence of cardiovascular diseases, hypertension, noninsulin dependent diabetes mellitus and dyslipidemia in the late adulthood are in connection with intrauterine retardation, characterized by low birth weight. One possible explanation of this phenomenon is the abnormality of hypothalamus hypophysis-adrenal cortex axis due to the accelerated growth. The authors investigated the steroid levels of young adults; whom birth weight were under 2500 g, and examined the relationship between hormone levels and some parameters of glucose metabolism and cardiovascular system. 75 subjects (43 female and 32 male patients, mean age: 19.6 and 19.8 years, respectively; range 18-22 ys) with low birth weight and without any sign of chronic disease, and 30 healthy, age matched controls with normal birth weight were investigated. The basal serum cortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate (DHEAS), androstenedione (AD), 17-hydroxyprogesterone (17OHP), estradiol (OE), sex-hormone binding globulin (SHBG), FSH, LH and insulin levels were determined. Moreover, oral glucose tolerance test with 75 g glucose (OGTT), impedance cardiography as well as ambulatory blood pressure monitoring were done by all subjects. In both sexes in subjects with low birth weight the mean serum cortisol level was significantly higher, than in the normal controls. In female patients the serum DHEA, DHEAS, AD, and 17OHP levels were significantly higher than in the controls. Moreover, among these females a relationship was found between the elevations of adrenal and gonadal steroids and hyperinsulinemia, characterized by increased insulin response during OGTT. In male subjects a significant correlation was found between serum cortisol levels and systolic blood pressure and heart rate. In females there was a positive relationship between serum DHEA and heart rate. Summarized, the basic abnormality in patients with low birth weight seems to be a relative hypercortisolism, and in females because of hyperinsulinemia exists a mild hyperandrogenism as well. The hypercortisolism may cause cardiovascular abnormalities in males directly, while in females indirectly through the hyperinsulinemia and hyperandrogenism. These subtle abnormalities can be detected when no clinical signs present themselves, in young adulthood, giving the opportunity of taking preventive actions. PMID- 11031834 TI - [Balloon dilatation in the management of gastric outlet stenoses: long-term results]. AB - The authors introduced dilatation of postoperative gastric outlet stenoses in 1981 and later in peptic, corrosive and postvagotomic gastric outlet stenoses. They performed gastric outlet stenoses dilatation with balloon catheter 153 times on 66 patients between September 1981-1999. On three of 8 malignant cases--who had dilatation because of recidive inoperable tumor--metal prosthesis implantation had been performed and others underwent surgery. On 58 patients who suffered from benign stenoses 144 balloon catheter dilatation had been done. After dilatation during the follow up time at 39/58 cases (67.2%) there was no need for operation. The stenoses of anastomosis after antrectomy and pyloric preserving pancreatoduodenectomy at 15 cases had been cured by dilatation. At pyloric and bulbus stenoses two third of the patients, 14 from 21 (67%) and at the cases of corrosive gastric outlet stenoses 6 of 17 (35%) had been treated successfully by dilatation. At postvagotomic functional stenoses 80% of the patients were treated successfully by 23 mm diameter balloon catheter. The dilatation has no long-term effect in malignant stenoses so it was used just for diagnosis or in rare inoperable cases for the preparation of prosthesis implantation. After dilatation therapy they detected one case of artery hemorrhage and two cases of perforation. The balloon catheter dilatation of the gastric outlet stenoses is a very important diagnostic and endoscopic therapeutic method which--depending on the cause--makes operation avoidable in 66% (35-100) of the benign stenoses. PMID- 11031835 TI - [The role of bacteriology in the treatment of tuberculosis; the Hungarian program]. AB - A positive result of the bacteriological examination of Mycobacterium tuberculosis is signal to start treatment. The response to antituberculosis chemotherapy in patients with positive bacteriology is best evaluated by repeated examinations. The recommendation of the Hungarian National Tuberculosis Programme for the initial intensive phase of chemotherapy of previously untreated patients is based on the frequency of primoresistance determined by the National Reference Laboratory. The treatment of multidrug resistant tuberculosis can be supported by the result of previous susceptibility test and/or by the outcome of multicenter validated Bactec 460 susceptibility test of second-line antimicrobial drugs. The presence of a mutation within a gen is predictive of rifampicin resistance of M. tuberculosis. The results are available within 48 h. 92.4% of rifampicin resistant and nearly 100% of pyrazinamid resistant isolates in the Reference Laboratory were resistant to isoniazid. On the other hand streptomycin resistant strains were susceptible to amikacin in a rate of 38:1. Some rifampicin resistant strains are susceptible to rifabutin. The Reference Laboratory observed among the first that some ofloxacin resistant strains are susceptible to ciprofloxacin. It was the first which demonstrated effectivity of amikacin in the treatment of pulmonary tuberculosis. It was drawn attention to the importance of the PAS infusion therapy of multidrug resistant tuberculosis cases by observing bacteriological conversion. PMID- 11031836 TI - [Memory of Ignac Semmelweis in Italy at the end of the 19th century]. PMID- 11031837 TI - [The magic snake. Facts from a symbol's history]. PMID- 11031838 TI - [Some novel therapies for multiple sclerosis. 1950]. PMID- 11031839 TI - [Should consensus techniques be included with qualitative investigation techniques?]. PMID- 11031840 TI - [Fascination with Popper in contemporary epidemiology]. PMID- 11031841 TI - [Popper and the problem of induction in epidemiology]. AB - In this article we are discussing a few of the contributions by the Austro British philosopher Karl R. Popper, one of our most influential contemporary thinkers, whose epistemological and socio-political theories have also penetrated the sphere of epidemiology. We are focusing mainly on the so-called problem of induction. We sustain, in line with Popper, that the scientific method does not use inductive reasoning, but rather hypothetical-deductive reasoning. Although the movement from the data evaluating a hypothesis to a conclusion on the latter goes from the specific to the general, that is, in an inductive direction, the induction does not exist as a reasoning process or inference. That is, there is no method that enables us to infer or to verify hypotheses or theories (we cannot explore all of the possible situations to see whether the theory stands up), or even to render them very probable. Besides, scientists look for highly informative theories, not highly probable ones. What we actually do is to propose a hypothesis as a tentative solution to a problem, to confront the prediction deduced from the hypothesis with actual experience, and evaluate whether the hypothesis is rejected or not by the facts. As theories cannot be verified, we can only accept them if they withstand an attempt to reject them. Consequently, the test of a theory consists of criticism or a serious attempt at falsification, that is, the elimination of error within a theory, in order to reject it if it is false. The objective is, thus, the search for true theories. For this purpose, the scientific method uses a systematic set of methodological (not logical) rules, that is, decisions. These methodological rules or principles can be summed up in two: [symbol: see text]be inventive and critical!, that is, propose bold hypotheses and subject them to severe tests of experience. Logic plays its role mainly by allowing us to deduce from a hypothesis the predictions to be confronted with the facts or evidence. This is applicable both to statistical inference as well as to causal inference. We argue that the criteria of causality used in epidemiology are none other than rules of the method designed for the same purpose: they are concerned with eliminating or reducing errors (chance, bias...) on testing a causal hypothesis. Consequently, the so-called ausal inference, the step from evidence to causal theory, is not a logical inductive or probabilistic process but rather a decision based on the evaluation of a causal hypothesis thanks to methodological rules such as the criteria of causality. We believe that the interest of the debate between the Popperian and the inductivist epidemiologists is not merely a matter of words, as, if we are aware that we do not operate inductively, that we cannot establish firmly hypotheses, not even affirm them probabilistically, we will presumably adopt a humbler attitude and look more for the errors in our theories than for their facile examples of confirmation. PMID- 11031842 TI - [Comparison between several standard populations for age-adjusting]. AB - BACKGROUND: To analyse the effect the choice of reference population has on different indicators derived from mortality rates age-adjusting. METHODS: The mortality rates for different causes of death in Spain from 1971 to 1992 were adjusted by using four reference populations: Spanish populations from 1971 and 1992, the Standard European population and the standard world population. The results obtained with the four populations were compared by applying three indicators: difference between the rates of 1992 and 1971, ratio between the rates of 1992 and 1971 and annual percentage change between 1971 and 1992. RESULTS: In the majority of the causes of death studied, including the total, ischemic heart disease and almost all of the tumours, the ratio between the rates and the percentage of annual change are similar regardless of the standard population used. However, the difference in rates is very noticeable in relation to the reference population. In the infectious diseases and testis cancer, the opposite occurs: the different in rates is steadfast while the ratio between the rates varies with the standard population. Finally, the mortality due to pneumonia, Parkinson's disease and encephalic cancer show changes all three indicators used. CONCLUSIONS: An analysis of the specific rates by age must be made before proceeding with their adjustment. This analysis will enable us to ascertain whether the adjustment by age is correct and which indicator (difference, ratio or percentage change) will be appropriate for making comparisons. PMID- 11031843 TI - [Use of non-conventional amphotericin B at the San Carlos Clinical Hospital]. AB - BACKGROUND: Amphotericin B is the treatment of choice for systemic fungal infections, however, its clinical usefulness is limited by its toxicity. The lipid formulations appear to be equally effective and safer, but are more costly. The increase in the consumption of, and expenditure on these formulas led us to undertake a study in order to identify their profile of use (quantitative and qualitative) and to assess the financial repercussions when used inappropriately. METHODS: A set of rules were developed for the use of amphotericin B, and the quality of the prescription of non-conventional amphotericin B (amphotericin B notC) was evaluated retrospectively together with the financial repercussions of its inappropriate use. RESULTS: In 54% of the treatments studied, a poor selection of amphotericin B was made; in 3.5%, the use of amphotericin B was not indicated. The excess expenditure derived from the inappropriate use amounted to 42 million pesetas, 35% of the total expenditure on medicines; the expenditure due to unnecessary prescription was 1,720,327 pesetas. CONCLUSIONS: The retrospective evaluation has shown that there is a high percentage of treatments that do not conform with the recommendations contained in the prescription rules. The holding of information sessions would assist in achieving a more efficient selection of the amphotericin B notC; this would improve prescription quality, which might also deliver significant financial savings. PMID- 11031844 TI - [Prevalence of tuberculosis infection prevalence in a provincial prison]. AB - BACKGROUND: Tuberculosis in prisons is a serious public health problem. In those penitentiaries with a high prevalence of co-infection by M. tuberculosis and HIV infection, the treatment of the tubercular infection can be an important tool for prevention and control. Ascertaining the prevalence of tubercular infection, the access and adherence to the treatment of tubercular infection in a prison population without a history of treatment of tubercular disease or infection could be of great interest in order to assess the efficacy of this measure. METHODS: A cross study was made of the population confined at 28 June 1999 in a Penitentiary with a programme of prevention and control of tuberculosis, based on the directly observed therapy (DOT) of the patients, on early diagnosis and the treatment of the tubercular infection. The treatment of the tubercular infection was prescribed following individualised counselling, reaching an agreement with the patient as to the modality of treatment (DOT, daily or twice-weekly check up). The clinical histories of the patients were reviewed in order to determine the prevalence of patients infected by M. tuberculosis, of those who had completed treatment and of those who were still undergoing treatment of tubercular infection. The Eidus-Hamilton test was performed in the case of patients under treatment for tubercular infection in order to evaluate compliance. RESULTS: Of the 219 inmates, 127 (58%) presented criteria of tubercular infection. Treatment of the tubercular infection was indicated in 113 people of whom 29 refused (25.7%). Of the 84 patients who initiated treatment for their tubercular infection, 22 (26.2%) abandoned it, 39 (46.4%) had finalized the treatment, and 23 (27.4%) were still undergoing treatment. Of the latter, 95.7% showed positive to the Eidus-Hamilton test. 48% of those infected by M. tuberculosis, either had finalized or were undergoing the treatment against tubercular infection correctly. CONCLUSION: The prevalence of tubercular infection observed in this group is very high. Acceptable levels of access and adherence to the treatment of tubercular infection have been observed, which translates into a high level of treatment coverage of this infection in the population studied. PMID- 11031846 TI - [Situation of the epidemic strain C:2b:P1.2,5 of Neisseria meningitidis after A+C vaccination]. AB - Following the use of an extensive immunisation campaign targeting the population between ages 2 and 19 in the majority of the Autonomous Communities (Regions), for the purpose of controlling the "outbreak or epidemic wave" caused by serogroup C in 1996-97, there was great interest in ascertaining the situation of the well-known epidemic strain C:2b:P1.2.5, which had been identified as the cause of the changes brought about in the epidemiological profile of meningococcal disease in Spain, as this would enable us to analyse some of the possible future tendencies of this disease in our country. An analysis of the situation in the first 10 months of 1999 reveals that we have reverted to figures similar to those which, both insofar as the number of cases as well as the frequency of serogroups, were observed in Spain at the beginning of the decade of the nineties, although the epidemic strain C:2b:P1.2.5 represents 56% of the total cases of serogroup C. The strains of serotype 2a, very frequent in serogroup C in European countries, continue to represent a minority in Spain. Finally, it is of interest to mention the appearance of antigen variants which could be the result of processes of intra-specific genetic recombination and which would presumably have been selected in terms of their evolutive advantages. PMID- 11031845 TI - [Meningococcal disease. The situation in Spain in the 1998-1999 period]. AB - BACKGROUND: The change in the epidemiological pattern undergone by meningococcal disease in a large part of the country in the 1996-1997 season and the decision to intervene by means of a mass vaccination campaign in the age group between 18 months and 19 years of age, using a bivalent polysaccharide vaccine, justified to enhance the epidemiological surveillance of this disease. METHODS: Rates and other indicators of incidence and mortality were calculated for the 1998-1999 campaign by serogroup and age, according to the data notified to the Compulsory Disease Reporting System. RESULTS: The overall incidence was higher than the previous season, although, the rate ratio did not indicate a significantly greater risk. A statistically significant increase in the rate of incidence caused by serogroup B was registered, and for the second consecutive year, the serogroup C rate dropped, although not significantly. There was an increase in the number of deaths. The fatality rate due to serogroup C increased in the group between 1 and 4 years of age. The incidence remained high and with a clear seasonal pattern in the Communities that did not vaccinate. In the remainder, the incidence dropped and the seasonal pattern disappeared. CONCLUSIONS: Two years after the campaign, the predominance of serogroup B cases is generalised in all of the Autonomous Communities, except in the three where the immunisation was not performed. In the latter, in the course of the last two years, serogroup C has predominated. In the Communities where the vaccination campaign was carried out, there is an increase in the incidence, although not significantly, among children under 4 years of age. PMID- 11031847 TI - [Results of a study on the under-detection of meningococcus in vaccinated subjects in Galicia]. AB - BACKGROUND: On the basis of active surveillance and the monitoring of Meningococcal Disease (MD) following the vaccination campaign carried out in Galicia, it was observed that the proportion of isolations of the serogroups responsible for the disease among individuals suspected of Meningococcal Disease (SMD) who had been vaccinated was lower than among unvaccinated individuals. In view of this situation, a study was made in order to determine whether in the origin of those SMDs that were not isolated, we would find N. Meningitidis serogroup C, and to quantify the significance of the sub-detection of same. METHODS: For this purpose, and during the period under study (from the 26th week of 1997 to the 14th week of 1999), blood and cephalorachidian fluid samples were taken from the SMDs without isolation for their study with C protein reagent for type and serogroup. The analysis of the samples was performed by the microbiology laboratory of the Clinical Hospital of Santiago de Compostela. RESULTS: Of the 120 cases notified during the period under study, 65 were analysed by C protein reagent (38 vaccinated and 27 unvaccinated), with a positive reading for N. meningitidis in 65% (42 samples) 74% in vaccinated individuals and 52% in unvaccinated. By estimating, on the basis of the cases studied, the results for the total, and excluding the C protein reagent negative cases, we find that, for serogroup C, in only 27% of the cases occurring in vaccinated individuals was it possible to isolate it, in comparison with 80% in the case of unvaccinated subjects (p < 0.0001). These percentages are, in the case of serogroup B, 59% and 71%, respectively, a difference which is not statistically significant. CONCLUSIONS: The vaccine brought about an true sub-detection of serogroup C meningococci in the vaccinated cases. PMID- 11031848 TI - [Meningococcal disease in Spain, 1990-1997. Change in its epidemiological pattern]. AB - BACKGROUND: In Spain, the presentation of meningococcal disease in the 1980s was closely linked to the predominance of N. Meningitidis serogroup B. This situation changed at the beginning of the 1990s, observing a growing trend of serogroup C strains in the isolation procedures carried out in some areas of the country, together with an increase in the incidence of the disease starting in the 1995 1996 season. The purpose of this study is to characterise the epidemiological pattern of meningococcal disease in Spain during the 1990-97 season and, in particular, where the disease was caused by the phenotype C:2b:P1.2.5 in this period of etiological change. METHODS: Retrospective study of the cases of meningococcal disease in Spain, with the exception of the Autonomous Communities of Andalusia, Cantabria, Madrid, Basque Country and Valencia, between 1990-1997. The epidemiological data were obtained from the Compulsory Disease Reporting system, through individualised notification of cases, and the microbiological data from the Meningococcus Reference Laboratory of the National Microbiology Centre. RESULTS: The incidence of meningococcal disease in the period studied, 1990-1997, was 3.81 x 10 - 6 person-years, increasing by 0.1851 cases per 100,000 inhabitants per year. Starting in 1995, the incidence caused by serogroup C practically tripled with respect to the preceding period, with a rate of incidence in 1997 of 2 cases per 100,000 inhabitants. This increase was to a large extent due to the emergence of the phenotype C:2b:P1.2,5, which in that same year registered an incidence of 1 case per 100,000 inhabitants. The increase in incidence affected all age groups, but was particularly significant in the 5 19 year-old group (annual rate of growth: 13.3%; P < 0.001). It was also found that there was an increase in the number of outbreaks reported and cases associated to them. The overall mortality rate was 7.7% (95% confidence interval: 7.0-8.4), and the mortality associated with the phenotype C:2b:P1.2.5 was significantly higher than that of the serogroup B chosen as reference (odds ratio: 1.69; 95% CI: 1.05-2.71), following adjustment for age, sex, clinical form and year. CONCLUSIONS: The pattern of meningococcal disease in the territory studied, during 1990-1997, was characterised by an increase in the incidence of serogroup C, particularly the emerging phenotype C:2b:P1.2,5. Together with this increase, a displacement of the incidence to higher age groups was observed, together with a tendency towards temporospatial aggregations of cases and an increase in the mortality associated with the new phenotype. This pattern is characteristic of epidemic situations of meningococcal disease caused by serogroup C. PMID- 11031850 TI - [Evaluation of the vaccination campaign in Cantabria 2 years of epidemiological evolution]. AB - The results of the meningococcal vaccination campaign carried out in Cantabria in February-March 1997 are presented. In the short term, the campaign reduced the rate of incidence in the risk group from 21.33/100,000 in the previous 12 months, to zero in the 12 months following the campaign. In the medium term (2 and a half years later), the rate of incidence was maintained 80% lower in the risk group and 77.3% lower in the population overall. The vaccinal efficacy in the third year following the campaign remained above 91% for all age groups, although some biases commented upon in the study have meant that this efficacy has been underestimated. Also, the vaccinal efficacy is much higher than the percentages of seroprotection measured by the rate of bactericidal antibodies, particularly in the younger age groups. The clinical cases in vaccinated individuals experienced a benign clinical course. PMID- 11031849 TI - [Evolution of meningococcal disease in the community of Madrid. Effectiveness of antimeningococcal A+C vaccination]. AB - BACKGROUND: In 1997 (between 22 September and 14 November) an A + C meningococcal mass vaccination campaign was carried out in Madrid, targeting the age group of from 18 months to 19 years of age, in the face of an increase in the number of cases of meningococcal disease caused by serogroup C occurring in the 1996-97 season. This study forms a part of the impact assessment of that campaign. METHODS: The evolution of the meningococcal disease, by means of the comparison of rates of incidence; and the efficacy of the vaccination campaign was determined after one year (1997-98 season) and after two years (1997-98 and 1998 99 seasons) of monitoring. The vaccine efficacy has been calculated as [1 (Incidence rate in vaccinated/Incidence rate in unvaccinated)]* 100. RESULTS: A significant drop was registered in the incidence of serogroup C meningococcal disease on comparing the 1997-98 and 1998-99 seasons with the epidemic season (1996-97). The vaccine efficacy after two years of monitoring subsequent to the vaccination campaign was 76.9% for the global population between 18 months and 19 years of age and 88.5% in the group of vaccinated individuals between 15 and 19 years of age. CONCLUSIONS: The vaccine efficacy obtained is compatible with that described in the relevant literature. The significant reduction in the incidence of meningococcal disease caused by serogroup C was due to the vaccine efficacy obtained. PMID- 11031851 TI - [Surveys on the rates of healthy carriers of Neisseria meningitidis and charaterization of circulating strains]. AB - BACKGROUND: During the epidemic season of 1996-97, the increase in the incidence of Meningococcal Disease caused by N. Meningitidis serogroup C led to the initiation of vaccination programmes and to the performance of a number of studies, among which, two were designed to ascertain the prevalence of healthy carriers and are presented here. METHODS: Healthy carriers in the population at large: cross survey performed between March and May 1998, among residents in Extremadura. We took two age groups: 0-19 years of age, vaccinated (n = 1,140) and 20 years of age and over (n = 1,193), unvaccinated. Healthy carriers in a military barracks: cross survey on a military base in May 1998 (n = 619), taking two groups: < 20 years of age (n = 453) vaccinated, and 20 years of age and over (n = 166) unvaccinated. RESULTS: Healthy carrier in the population at large. The percentage of carriers for all of the serogroups was: 0-19 years of age 7.98, 20 years of age and older 3.32, total 4.60, for serogroup B: 4.43, 2.25 and 2.84; for C: 0.04, 0.10 and 0.08 respectively. The differences between the two age groups in serogroup B and the total are statistically significant, as well as the greater prevalence of B over C. The subtype of the three samples with isolation of serogroup C was: one strain NT:P1.6, another strain 2b:P1.2, and one that could not be sub-typed. A strain that could not be classified into a serogroup was sub-typed as 2b:P1.5. Healthy carriers in a military barracks. The percentage of carriers for all of the serogroups was: < 20 years 8.43, 20 years of age and over 6.84, total 7.27, for serogroup B: 4.82, 3.75 and 4.04; and for C: 0.60, 0.00 and 0.16 respectively. The higher prevalence obtained in military subjects in comparison with the population at large revealed statistically significant differences. Also, the prevalence of serogroup B was significantly higher than C. The only isolated C strain was sub-typed as: p1.2,5. CONCLUSIONS: The prevalence of healthy carriers of N. meningitidis in the population at large was 4.60%, being higher in the lower age group and the barracks. Very low prevalence of serogroup C. Impossible to conclude that the strain N. meningitidis serogroup C was circulating during the 1997/98 season. PMID- 11031852 TI - [Survey of carriers of Neisseria meningitidis in the health area of Gran Canary]. AB - BACKGROUND: A) To ascertain the rate of carriers and the Types of Neisseria Meningitidis circulating in the population resident in the health jurisdiction of Gran Canaria. B) to ascertain the pattern of distribution of such carriers. METHODS: A descriptive transversal design was made, with a random sampling in multiple stages and by conglomerates. A minimum sample size was determined at 707 individuals for an expected prevalence of 8.6%, with a rate of reliability of 95.6% and a precision of 0.02. Assuming that 15% if the individuals would not be willing to co-operate, the sample size was increased to 831 individuals, distributed in each conglomerate in proportion to the existing population. This size was distributed in turn into four groups by age and sex, in proportion to their significance in each basic health care zone selected at random. The individuals in the sample were identified from among those who attended the blood extraction units, and after they had passed the criteria of exclusion, their co operation was requested as volunteers in the study. If they accepted, a questionnaire was filled out with a number of variables of epidemiological interest and a pharyngeal smear was taken. Since the Primary Care units were selected on a simple random basis, and the same method was used to select the individuals within the units, the estimate of the prevalence was made by means of an unbiased estimator. RESULTS: A total of 828 samples were obtained, that is, 99.6% of the number foreseen. With the exception of three, all of the individuals selected participated voluntarily in the study, a circumstance rendering it highly representative. All of the strains obtained corresponded to N. Meningitidis Serogroup B, except for one identified as N. Meningitidis Serogroup C Sero/Subtype 4:P1.2,5. The strains of N. Meningitidis serogroup B identified corresponded to 25 different sero-subtypes. The prevalence determined after having studied the sample was 6.45%, the variance = 0.0275 and the standard error = 1.66. We can conclude with a 95% degree of reliability that the prevalence of carriers of N. Meningitidis in the Gran Canaria health jurisdiction is estimated to be between 3.2% and 9.7%. CONCLUSIONS: A clear predominance of N. Meningitidis serogroup B strains among carriers has been verified. There are no statistically significant differences in the prevalence observed among the different age groups nor between sexes. PMID- 11031853 TI - [Effectiveness and duration of the immunity of the vaccine against serogroup A+C meningococcus]. AB - BACKGROUND: In 1995 the Community of Madrid detected an increase in the number of cases of meningococcal disease caused by serogroup C. In 1997 a mass vaccination campaign was carried out in relation to the population between 18 months and 19 years of age. The purpose of this study is to ascertain the immune response produced by the vaccine and its relationship to the age of the subjects. METHODS: A sample group of 1,003 children vaccinated during the campaign was selected. A blood sample was extracted prior to vaccination and after one, six (only a under 5 years old) and twelve months had transpired. In order to assess the immune response, the levels of bactericidal and total antibodies were measured. RESULTS: The prevalence of seroconversion measured by bactericidal antibodies is 89.6%. The response is low in children under 3 (34.8%), increases with age and, from 7 years on, surpasses the 90% mark. After 6 months, the prevalence of protective levels in children under 5 years of age drops noticeably (31.3%). After one year, the prevalence drops significantly, particularly in children under 7 years of age. The proportion of individuals with total antibody response after one month is over 90%, and remains high after one year in all of the age groups (97.5%). CONCLUSIONS: The response measured by means of total antibodies contradicts the clinical response to the vaccination and the measurement by means of bactericidal antibodies underestimates the protection if it is compared with the results of vaccinal efficacy, for which reason, we need to search for biological indicators that would correlate adequately with the clinical response following immunisation. PMID- 11031855 TI - [Results of a serologic study after vaccination against serogroup C Neisseria meningitidis in children]. AB - BACKGROUND: An increase in the incidence of meningococcal disease in the Region of Murcia, Spain, during the 1996/97 season led the General Directorate for Health to carry out a Vaccination Campaign against the serogroup C meningococcus in September-October 1997. The purpose of this study was to ascertain the percentage of children under 5 years of age who revealed post-vaccinal seroconversion one month following vaccination, and the percentage who retained immunity after one year. METHOD: The monitoring of 296 children between 18 and 59 months of age. The serological determinations were performed prior to vaccination, one month following vaccination and after one year. The antibody titres were determined according to a "bactericidal test" in the Center for Disease Control. RESULTS: Of the 296 children studied, only 11 (3.7%) revealed bactericidal antibody titres prior to vaccination. After one month, of the children who did not show antibodies prior to vaccination, 167 (63.7%) seroconverted. A statistically significant linear tendency (p < 0.001) of increase in the percentage of seroconversion in relation to the vaccination age was observed. One year after vaccination, from among the children seroconverted one month following vaccination, only 6 (4.3%) revealed bactericidal antibodies. CONCLUSION: The percentage of seroconversion in children under 5 years of age, following administration of the capsular polysaccharide C vaccine, was seen to clearly increase with the age of the subjects. The seroprotection acquired in vaccinated children declined quickly during the year following vaccination. PMID- 11031854 TI - [Levels of bactericidal antibodies against meningococcus C after vaccination in 2 to 6-year-old children in Andalusia]. AB - BACKGROUND: In 1997, 18.5% of the cases of Meningococcal Disease caused b serogroup C in Andalusia were children between 2 and 4 years of age; ages where the initial immune response and the duration of the capsular A + C meningococcal polysaccharide vaccine is less than in older age groups. Research was designed in order to measure the immune response produced by this vaccine in children from 2 to 6 years of age and to compare it with the natural immunity present in unvaccinated children. METHODS: I. Dual monitoring study: a) groups of children vaccinated previously and control groups, b) groups of children who were going to be vaccinated, for pre and post-vaccination (1, 6 and 12 months) analysis and a control group. II. The bactericidal activity was measured according to the standardised protocol of the CDC with regard to the strain of N. meningitidis C 11. The sera with bactericidal activity (TAB) > 1:8 were considered to be protective. RESULTS: 1 and 2 months following vaccination, the proportion of TAB > 1:8 was significantly higher than that of the control group (65.6% and 73% in comparison to 2.2% and 12%). In the pre-vaccine and post-vaccine (after 6, 7, 12 and 13 months) verification, no significant difference between vaccinated individuals and controls was observed. CONCLUSIONS: The differences between vaccinated and unvaccinated individuals 1 and 2 months following vaccination indicate seroconversion in the vaccinated individuals. For the age group of between 2 to 6 years of age, the bactericidal activity acquired decline quickly, as, after 6 months, differences between this group and the control group are no longer observed. PMID- 11031856 TI - [What is your diagnosis? Localized, ulcerated plaque rupture of the descending thoracic aorta with intramural hematoma]. PMID- 11031857 TI - [Cholesterol determination in ambulatory general practice within the scope of the Brugg/AG quality circle]. AB - Findings of studies designated by the acronyms 45, WOSCOP, LIPID and CARE have provided data that led physicians to rethink their "cholesterol testing behavior." Twelve physicians participated in a study conducted in cooperation with the quality circle of the Brugg region in Aargau. Each doctor collected data from the files of 100 patients and filled in a questionnaire. Cardiovascular risk factors and arteriosclerotic secondary illnesses were compiled along with demographic data. Moreover, information was gathered on whether the patient's cholesterol levels had been checked within the last five years. In total, 1183 questionnaires were evaluated, comprising 691 women and 492 men with an average age of 48.6 years. Cholesterol levels had been determined within the last five years in 61.2%. The individual testing behavior of the physicians varied. One third tested all three blood lipid values (total cholesterol, HDL and triglyzerides) with an equivalent frequency. By contrast, one-third primarily only determined total cholesterol. The last one-third mostly tested total cholesterol and triglyzeride levels. The frequency of serum lipid tests increased proportionally to the number of risk factors. Cholesterol levels were tested less frequently (approx. 55%) in smokers and patients who never practiced sports than in patients with other risk factors (approx. 85%). The testing rate increased proportionally to the increasing number of arteriosclerotic secondary illnesses, but did not reach the one-hundred percent mark until 3 secondary illnesses were involved. Dietary counseling was the primary therapeutic intervention; medication was prescribed with a much rarer frequency. Although 75% of the patients with arteriosclerotic secondary illnesses had measurably higher cholesterol levels (> 5.2 mm/l), approx. one-third of the patients with coronary heart disease received therapy with cholesterol-lowering drugs as compared to only 16% of the patients with peripheral occlusive arterial disease. The conclusions that we can draw from these results for clinical practice are: if cholesterol levels are to be determined in order to evaluate a patient's cardiovascular risk, HDL cholesterol levels should also be tested since the ratio of these two values is a good predictor. To date, triglyzeride levels have not been identified as relevant risk factors for vascular events. Physicians evaluated the other risk factors variably, in particular with regard to obesity. For clinical practice, the potential risk factor of lack of exercise should be defined more accurately. In patients with clinically manifest arteriosclerotic secondary illnesses, serum lipids should be tested routinely and appropriate therapy induced when required. PMID- 11031858 TI - [Dementia--a genetic disease?]. PMID- 11031859 TI - [Human fascioliasis symptoms. A case report with review of the literature]. AB - We present the case of a 17 years old man with fasciola hepatica infection. After initial therapy with Praziquantel and Metronidazole abdominal pain as well as eosinophilia persisted. The diagnosis was made by ERC and examination of the bile, where typical ovulas were found. An analysis of faeces and the serology (ELISA, IFAT) were not diagnosticated. After papillotomy, extraction of the parasites and therapy with Triclabendazole the patient became asymptomatic, the serology turned to negative and the haematology was normalised. PMID- 11031860 TI - [Jugular lymph node metastasis of papillary thyroid gland carcinoma 10 years later]. AB - We present a patient with occult papillary thyroid carcinoma, in whom a single nodal metastasis had been excised. Ten years later a nodal recurrence was diagnosed by ultrasound and fine needle aspiration cytology. The diagnosis of recurrence was erroneous, and we discuss the differential diagnostic problems, the natural course, and the adequate treatment of occult papillary microcarcinoma. PMID- 11031861 TI - [Miserere. Incarcerated femoral hernia]. PMID- 11031862 TI - [Family anamnesis as the key to diagnosis.1. Multi-infarct syndrome in atrial secundum-type septal defect. 2. Thrombophilia of uncertain etiology. Homocysteine in the upper normal range]. PMID- 11031863 TI - [Differentiated approaches to the treatment of nonstabilized primary open-angle glaucoma with normalized intraocular pressure considering its pathogenic features]. AB - Clinical efficiency of dedystrophic treatments for nonstabilized primary open angle glaucoma (POAG) in the presence of normalized intraocular pressure is compared in 168 patients (246 eyes). In one group of patients ischemic angiopathy and hyperreactivity of optic vessel adrenoreceptors associated with hypokinetic central hemodynamics and constitutional metabolic status of the organism was corrected by cinnarisin and riboxin. Patients with predominating congestive angiopathy symptoms, hyper- or eukinetic circulation and "slow" acetylation were treated by pantothenic acid, endotelon, and hyperbaric oxygenation. In both groups epithalamine, eiconol, and magnetic laser therapy were used, if indicated. This helped stabilize the process in 91% patients with initial POAG, in 87.5% with well-developed condition vs. 66.1% and 38.2% patients treated by traditional therapy (period of observation 3 years). PMID- 11031864 TI - [Densitometric analysis of crystalline lens nucleus in the choice of strategy of surgical treatment of cataracts]. AB - Computer analysis of TV images of the eye was used for quantitative evaluation of optic density of the lens nucleus in patients with cataracts in need of surgical treatment. Indications to cataract phacoemulsification were determined using classification of the degree of the lens nucleus maturity. This classification is based on the maximal values of optic density of the anterior third of the nucleus, obtained by the lens densitometry, coefficient of light absorption by the lens nucleus, and tentative subjective classification of the lens nucleus maturity by a color scale. In initial cataract (first degree) the mean maximal optic density of the anterior third of the nucleus is 55.8 +/- 5.7 arb. units and coefficient of the lens nucleus light absorption is 0.93, in nuclear brown cataract (fourth degree) these values are 158.7 +/- 24.4 arb units and 1.81, respectively. Cataract with brown enlarged nucleus involving the cortical layers of the lens and high optic density of these layers, with lens nucleus light absorption coefficient more than 2.5 is a contraindication to phacoemulsification. PMID- 11031865 TI - [Changes in corneal topography after photorefractive keratectomy in myopia and their elimination]. AB - Topographic disorders in the cornea, regression of the refraction effect, and subepithelial corneal opacities are characteristic complications of photorefraction keratectomy. Disorders in corneal topography are early haze, asymmetrical haze, and optic zone decentration. Disorders in topography lead to decrease in visual acuity, numerous optic effects (monocular doubling, aureola, blurred image, etc.). Methods for these complications control by means of transepithelial ablation, which is carried out with due consideration for the data of topographic examinations under pseudofluorescence control, normalizes corneal topography in the majority of cases, notably increases visual acuity, decreases doubling, etc. No side effects were observed after transepithelial phototherapeutic keratectomy for a period of up to 12 months. PMID- 11031866 TI - [Combination of photorefractive and phototherapeutic keratectomy in the treatment of keratoconus]. AB - Twenty patients (28 eyes) with primary keratoconus of stages I-II (Amsler) were operated using photorefraction keratectomy (PRK) supplemented by phototherapeutic keratectomy (PRK). Mean UCVA before surgery was 0.04 +/- 0.004, BCVA 0.68 +/- 0.03. Eximer laser EC-5000(NIDEK) was used in the treatment. PRK included spherical, cylindrical ablations, or both. The diameter of ablation zone in PRK was 6 mm. in PTK the diameter of ablation zone was 8 mm, transition zone 9 mm. In PTK the ablation zone was decentered towards the cone apex (according to corneal computer topography). The method is patented in the Russian Federation. Visual acuity without correction notably improved in all 20 patients and its mean value after the operation was 0.73 +/- 0.03, 68% eye having visual acuity of 1.0. In one female patient keratoconus progressed 6 months after the operation. Haze was observed in 1 patient as a result of delayed reepithelialization. No progress of keratoconus was observed in other patients (period of observation 17.3 +/- 1.5 months). Hence, the new PRK-PTK method is an effective method for treating primary keratoconus of the first-second stage according to Amsler; the method arrests the progress of the condition in 96% cases. PMID- 11031867 TI - [Functional results of aphakia correction by different methods]. AB - Functional results of aphakia correction by different methods (eyeglasses, contact and intraocular lenses) are compared. The difference of this study from previous is that it was carried out in a group of patients to whom these methods were prescribed in succession. The rightfulness of such an approach to evaluation of functional results of various methods for aphakia correction is dictated by the fact that the norm is a relative parameter for many ophthalmic ergonometric studies, and thus the investigations need to be standardized (e.g., investigations in the same patients). Comparison of the efficiencies of different methods demonstrated that the best results are attained with intraocular lenses. PMID- 11031868 TI - [Mechanisms of sensory function recovery in partial optic nerve atrophy before and after treatment with ampli-pulse phoresis]. AB - Eighty-three patients (119 eyes) with partial optic nerve atrophy of different origin were treated by amplipulse phoresis and observed for 1 year. Based on the data of modern methods of examination of visual functions (changes in visual evoked potentials and achromatic static campimetry), the authors suggest that several mechanisms of visual function improvement underlie the therapeutic effect of amplipulse phoresis. These mechanisms are as follows: activation of the intact part of nerve fibers with restoration of myelin membranes, restoration of viability of nerve fibers in a state of parabiosis; a probable mechanism of visual acuity improvement is modification of the conduction of visual route on channels, but this hypothesis needs further investigations. PMID- 11031869 TI - [Effects of monochromatic light on ocular hydrodynamics]. AB - Effects of green and red optic filters on ocular hydrodynamics was studied. The results indicate that monochromatic green light causes a decrease of the true intraocular pressure, improves intraocular fluid discharge, and decreases Becker's coefficient. Green filters (ZSP) can be used as an additional method for conservative treatment of glaucoma patients. PMID- 11031870 TI - [Metabolic state of the other eye in post-traumatic uveitis]. AB - Biochemical analysis of the paired eye lacrimal fluid lipid peroxides (diene conjugates, Schiff's bases, TBA-active products), antioxidant enzymes (catalase, superoxide dismutase), dehydrogenases (lactate and malate dehydrogenases), and lactic and pyruvic acids was carried out during the early and remote posttraumatic period in 468 patients after perforating injuries to the eye. Signs of activation of lipid peroxidation, antioxidants, dehydrogenases, and of glycolysis intensification were detected. These changes corresponded to the results of analysis of lacrimal fluid of the injured eye and depended on the severity of injury, time elapsed after it, and severity and course of posttraumatic uveitis. The author claims that the detected metabolic shifts in the lacrimal fluid of paired eye are involved in the pathogenesis of posttraumatic uveitis. PMID- 11031871 TI - [Regularities of changes in parameters of humoral immunity in patients with eye diseases]. AB - Changes in the levels of natural antibodies in the lacrimal fluid, saliva, and blood immunoglobulin concentrations were studied in patients with ocular injuries, inflammatory (keratitis, scleritis, uveitis) and noninflammatory diseases of the eyes. These values can be used as additional parameters for the diagnosis and prediction of the treatment efficiency. The immune system functions as a universal system with different compartments, and therefore mathematical relationships between these parameters were studied. Positive linear regression and correlation relationships were detected between -log2 antibody titers in the serum and lacrimal fluid, antibody titers in the saliva and serum, and negative relationships between antibody titers in the lacrimal fluid and IgA concentration and between lacrimal antibody titers and IgA level. PMID- 11031872 TI - [Study of ocular hemodynamics and general somatic status of patients with primary open-angle glaucoma]. AB - Ocular hemodynamics and general somatic status were studied in 50 patients with primary open-angle glaucoma. Complex ophthalmological and therapeutic examinations were carried out, macro- and microcirculatory hemodynamics of the eye was examined, case histories and therapeutic status of patients were analyzed with due regard for previous local hypotensive and general vascular therapy. Revised recommendations for glaucoma patients aimed at correcting their life style include diet, regular vascular and metabolic therapy, normalization of orbital circulation in order to improve ocular hemodynamics and increase the tolerance of the optic nerve to intraocular pressure fluctuations. PMID- 11031873 TI - [Optic disk drusen]. AB - Optic disk drusen (ODD) is a much more prevalent condition than it is usually considered. It is important to recognize ODD in order to escape serious neurosurgical interventions and rule out the development of optic neuropathy leading to grave visual dysfunction. Three cases with ODD are described. The authors conclude that it is an acquired disease, which can be bi- and unilateral. Initial diagnosis "suspected congestive optic disk of both eyes" was erroneous in all cases and it was replaced by "pseudocongestion" because of ODD. A characteristic symptom of ODD is narrowing of visual field borders and presence of scotomas in the lower half of central visual field. Since ODD are a manifestation of progressive optic neuropathy leading to disorders in the central visual field, dedystrophic and (if necessary) ophthalmo-hypotensive therapy is to be carried out. PMID- 11031874 TI - [Irrigation system "Meturakol" in the treatment of dystrophic processes of the posterior eye segment]. AB - An original method for treating degenerative processes in the eye by means of Meturakol irrigation system is described. Studies were performed in 95 patients aged 60-70 years. Use of Meturakol irrigation system ruled out pain associated with frequent administration of drugs, ensured high concentrations of drugs in the posterior segment of the eye, and improved the efficiency of drug therapy in patients with degenerative diseases of the eyes. Good remote results are due to potent drug supply to ocular tissues and to the use of hemostatic sponge Meturakol, containing methyluracil. The method is offered for wide clinical use. PMID- 11031875 TI - [Characteristics of correction of myopia by photorefractive keratectomy in patients with presbyopia]. AB - Photorefraction keratectomy was performed with a Nidek EC-5000 excimer laser in 29 patients (58 eyes) with slight and medium myopia aged 40-60 years. After 12 months the mean visual acuity in the leading eye without correction was 0.82 +/- 0.12 in patients aged 40-50 (group 1) and 0.76 +/- 0.11 in those aged 51-60 years (group 2). The mean refraction of the leading eye in group 1 was -0.97 +/- 0.02 diopters and in group 2 0.72 +/- 0.04 diopters. In group 1 the resultant undercorrection was -1.6 +/- 0.4 diopters (107% of planned undercorrection) and in group 2 -1.8 +/- 0.4 diopters (72% of planned undercorrection). The proposed method effectively corrects slight and medium myopia in patients of presbyopia age. 82% patients were satisfied with the results of treatment: 67% of them did not need eyeglasses for reading and 54% could drive a car without eyeglasses. PMID- 11031876 TI - [Physiological mechanisms of the etiology of visual fatigue during work involving visual stress]. AB - Physiological parameters of vision were studied in three professional groups (a total of 1204 subjects): microscope operators, subjects working with magnifying glasses, and computer users. General and specific features of visual system fatigue formation were identified. Because of complete (in microscope operators) or partial (in subjects working with magnifying glasses and display users) "deprivation" of accommodation, these subjects develop early presbyopia (at the age of 30-35 years). In microscope operators long strain of accommodation system leads to professional myopia, while display users develop pseudomyopia. The highest overstrain is observed after 4 years of work in microscope operators, after 5 years in magnifying glass users, and after 6 years in computer users. PMID- 11031877 TI - [Laser scanning ophthalmoscopes: prospects of their use in ophthalmology]. PMID- 11031878 TI - [Clinical aspects, diagnosis and treatment of cyclotropia and torsion diplopia]. PMID- 11031879 TI - [X-linked juvenile retinoschisis]. PMID- 11031880 TI - [World initiative: elimination of avoidable blindness in the world]. PMID- 11031882 TI - [Hypopituitarism]. PMID- 11031881 TI - [Hypersecretion of growth hormone]. PMID- 11031883 TI - [Pituitary apoplexy]. PMID- 11031884 TI - [Syndrome of inappropriate secretion of antidiuretic hormone(SIADH)]. PMID- 11031885 TI - [Neuromuscular disorders associated with hyperthyroidism]. PMID- 11031886 TI - [Neuromuscular disorders associated with hypothyroidism]. PMID- 11031887 TI - [Neurological manifestation of hyperparathyroidism]. PMID- 11031888 TI - [Neurological manifestation of hypoparathyroidism]. PMID- 11031889 TI - [Pseudohypoparathyroidism]. PMID- 11031891 TI - [Adrenocortical insufficiency, adrenal crisis]. PMID- 11031890 TI - [Neurological symptoms associated with hypercortisolism]. PMID- 11031892 TI - [Primary aldosteronism]. PMID- 11031893 TI - [Pancreatic endocrine tumor]. PMID- 11031894 TI - [Multiple endocrine neoplasia syndrome(MEN)]. PMID- 11031895 TI - [Diabetic peripheral neuropathy]. PMID- 11031896 TI - [Diabetic autonomic neuropathy]. PMID- 11031897 TI - [Diabetic amyotrophy]. PMID- 11031898 TI - [Pseudotabes diabetica]. PMID- 11031899 TI - [Diabetic coma]. PMID- 11031900 TI - [Thiamine deficiency]. PMID- 11031901 TI - [Pyridoxine deficiency and dependency]. PMID- 11031902 TI - [Niacin deficiency disease(pellagra)]. PMID- 11031903 TI - [Neurology of vitamin B12 deficiency]. PMID- 11031904 TI - [Folate deficiency and congenital folate malabsorption]. PMID- 11031905 TI - [Vitamin A deficiency and neurological findings]. PMID- 11031906 TI - [Vitamin D deficiency]. PMID- 11031907 TI - [Vitamin E deficiency]. PMID- 11031908 TI - [Vitamin K deficiency]. PMID- 11031909 TI - [Pyridoxine toxicity]. PMID- 11031910 TI - [Vitamin A toxicity and neurological findings]. PMID- 11031911 TI - [Vitamin D toxicity]. PMID- 11031912 TI - [Hepatic encephalopathy [acute type, chronic/recurrent type and mixed/terminal coma type]]. PMID- 11031913 TI - [Portal-systemic encephalopathy or shunt encephalopathy]. PMID- 11031914 TI - [Hepatic myelopathy, hepatic neuropathy]. PMID- 11031915 TI - [Chronic acquired(non-Wilsonian) hepatocerebral degeneration(CAHD)]. PMID- 11031916 TI - [Dialysis encephalopathy, dialysis dementia]. PMID- 11031917 TI - [Dialysis dysequilibrium syndrome(DDS)]. PMID- 11031918 TI - [Syndrome related to chronic dialysis]. PMID- 11031919 TI - [Carpal tunnel syndrome in hemodialysis patients]. PMID- 11031920 TI - [Renal tubular acidosis]. PMID- 11031922 TI - [Hypercapnic coma(CO2 narcosis)]. PMID- 11031921 TI - [Neurologic disorders in renal transplantation]. PMID- 11031923 TI - [Hypoxic encephalopathy]. PMID- 11031924 TI - [Pickwickian syndrome and sleep apnea syndrome]. PMID- 11031925 TI - [Hyperventilation syndrome]. PMID- 11031926 TI - [Neurological manifestations in patients with hematopoietic stem cell disease]. PMID- 11031927 TI - [Abnormality of red blood cells]. PMID- 11031929 TI - [Abnormality of lymphocytes]. PMID- 11031928 TI - [Granulocytic disorder(except malignant tumor)]. PMID- 11031930 TI - [Immunodeficiency syndromes]. PMID- 11031931 TI - [Neurological manifestations of platelet disorders]. PMID- 11031932 TI - [Hemorrhagic diathesis due to coagulative and fibrinolytic abnormalities]. PMID- 11031933 TI - [Thrombogenic diseases]. PMID- 11031934 TI - [Neurologic syndrome due to hematologic malignancy]. PMID- 11031935 TI - [Pathological serum proteins(M-components)]. PMID- 11031936 TI - [The syndrome of the sea-blue histiocyte]. PMID- 11031937 TI - [Neurological disorders of light chain deposition disease]. PMID- 11031938 TI - [The symptoms and pathogenesis of chronic GVHD]. PMID- 11031939 TI - [Neurological manifestations in patients with hyperviscosity syndrome]. PMID- 11031940 TI - [Thrombotic microangiopathy in hematopoietic stem cell transplantation]. PMID- 11031941 TI - [Cryoglobulinemia]. PMID- 11031942 TI - [Systemic lupus erythematosus]. PMID- 11031943 TI - [Rheumatoid arthritis, malignant rheumatoid arthritis]. PMID- 11031944 TI - [Systemic scleroderma]. PMID- 11031945 TI - [Polymyositis and dermatomyositis]. PMID- 11031946 TI - [Rheumatic fever]. PMID- 11031947 TI - [Neural disorders in mixed connective tissue disease(MCTD) and overlap syndrome]. PMID- 11031948 TI - [Sjogren's syndrome: involvement of peripheral and central nervous system]. PMID- 11031949 TI - [Polyarteritis nodosa and microscopic polyangiitis]. PMID- 11031950 TI - [Allergic granulomatous angitis(Churg-Strauss syndrome)]. PMID- 11031951 TI - [Polyangiitis overlap syndrome]. PMID- 11031952 TI - [Wegener's granulomatosis]. PMID- 11031953 TI - [Temporal arteritis]. PMID- 11031954 TI - [Takayasu arteritis(Takayasu disease, pulseless disease)]. PMID- 11031955 TI - [Henoch-Schonlein syndrome]. PMID- 11031956 TI - [Cutaneous leukocytoclastic angiitis(hypersensitivity vasculitis)]. PMID- 11031957 TI - [Kawasaki disease]. PMID- 11031958 TI - [Granulomatous angiitis of the central nervous system(GANS), isolated CNS vasculitis]. PMID- 11031959 TI - [Thromboangitis obliterans(TAO), Buerger disease]. PMID- 11031960 TI - [Behcet's disease]. PMID- 11031961 TI - [Cogan syndrome]. PMID- 11031963 TI - [Paraneoplastic cerebellar degeneration]. PMID- 11031962 TI - [Paraneoplastic limbic encephalitis]. PMID- 11031964 TI - [Paraneoplastic brainstem encephalitis]. PMID- 11031965 TI - [Paraneoplastic opsoclonus myoclonus syndrome]. PMID- 11031966 TI - [Paraneoplastic retinal degeneration, cancer-associated retinopathy]. PMID- 11031967 TI - [Paraneoplastic necrotizing myelopathy]. PMID- 11031968 TI - [Paraneoplastic motor neuron disease]. PMID- 11031969 TI - [Stiff-man(person) syndrome]. PMID- 11031970 TI - [Paraneoplastic sensory neuropathy]. PMID- 11031971 TI - [Paraneoplastic Crow-Fukase syndrome]. PMID- 11031972 TI - [Paraneoplastic subacute motor neuropathy]. PMID- 11031973 TI - [Paraneoplastic dermatomyositis]. PMID- 11031974 TI - [Lambert-Eaton myasthenic syndrome(LEMS)]. PMID- 11031975 TI - [Inherited neurological diseases due to carbohydrate metabolic defect]. PMID- 11031976 TI - [Hyperphenylalaninemia]. PMID- 11031977 TI - [Hyperprolinemia]. PMID- 11031978 TI - [Disorders of the urea cycle]. PMID- 11031979 TI - [Maple syrup urine disease]. PMID- 11031981 TI - [Carnosinemia]. PMID- 11031982 TI - [Organic acidemia, inborn errors of metabolism]. PMID- 11031980 TI - [Homocystinuria]. PMID- 11031983 TI - [Inborn errors of purine metabolism]. PMID- 11031984 TI - [Inborn errors of pyrimidine metabolism]. PMID- 11031985 TI - [Disorder of DNA repair]. PMID- 11031986 TI - [Inborn errors of lipoprotei and lipid metabolism]. PMID- 11031987 TI - [Porphyrias]. PMID- 11031988 TI - [Inborn errors of Metal metabolism]. PMID- 11031989 TI - [Vitamin dependency syndrome]. PMID- 11031990 TI - [Inborn errors of bilirubin metabolism]. PMID- 11031992 TI - [Hurler syndrome(MPS IH), Scheie syndrome(MPS IS)]. PMID- 11031991 TI - [Peroxisomal disorders]. PMID- 11031993 TI - [Hunter syndrome]. PMID- 11031994 TI - [Sanfilippo syndrome]. PMID- 11031995 TI - [Morquio syndrome, beta-galactosidase deficiency]. PMID- 11031996 TI - [Maroteaux-Lamy syndrome]. PMID- 11031997 TI - [Sly disease]. PMID- 11031998 TI - [Sphingolipid activator protein deficiency]. PMID- 11031999 TI - [I-cell disease and pseudo-Hurler polydystrophy]. PMID- 11032000 TI - [Niemann-Pick disease type C]. PMID- 11032001 TI - [Sialic acid storage disease]. PMID- 11032002 TI - [Cystinosis]. PMID- 11032003 TI - [Neuronal ceroid lipofuscinosis (NCL, Batten's disease)]. PMID- 11032004 TI - [Lowe syndrome(oculocerebrorenal syndrome of Lowe; OCRL)]. PMID- 11032005 TI - [Lysosomal glycogen storage disease without acid maltase deficiency(Danon disease)]. PMID- 11032006 TI - [CDG syndrome(congenital disorders of glycosylation)]. PMID- 11032007 TI - [Mitochondrial disease]. PMID- 11032008 TI - [Ehlers-Danlos syndrome]. PMID- 11032009 TI - [Osteochondrodysplasia]. PMID- 11032010 TI - [Marfan syndrome]. PMID- 11032011 TI - [Hereditary amyloidosis]. PMID- 11032012 TI - [Norrie disease]. PMID- 11032013 TI - [Inborn errors of catecholamine metabolism]. PMID- 11032014 TI - [Alexander's disease]. PMID- 11032015 TI - [Pelizaeus-Merzbacher disease(PMD)/spastic paraplegia 2(SPG2)]. PMID- 11032016 TI - [Sudanophilic leukodystrophy]. PMID- 11032017 TI - The c-Myc-interacting adaptor protein Bin1 activates a caspase-independent cell death program. AB - Cell death processes are progressively inactivated during malignant development, in part by loss of tumor suppressors that can promote cell death. The Bin1 gene encodes a nucleocytosolic adaptor protein with tumor suppressor properties, initially identified through its ability to interact with and inhibit malignant transformation by c-Myc and other oncogenes. Bin1 is frequently missing or functionally inactivated in breast and prostate cancers and in melanoma. In this study, we show that Bin1 engages a caspase-independent cell death process similar to type II apoptosis, characterized by cell shrinkage, substratum detachment, vacuolated cytoplasm, and DNA degradation. Cell death induction was relieved by mutation of the BAR domain, a putative effector domain, or by a missplicing event that occurs in melanoma and inactivates suppressor activity. Cells in all phases of the cell cycle were susceptible to death and p53 and Rb were dispensable. Notably, Bin1 did not activate caspases and the broad spectrum caspase inhibitor ZVAD.fmk did not block cell death. Consistent with the lack of caspase involvement, dying cells lacked nucleosomal DNA cleavage and nuclear lamina degradation. Moreover, neither Bcl-2 or dominant inhibition of the Fas pathway had any effect. In previous work, we showed that Bin1 could not suppress cell transformation by SV40 large T antigen. Consistent with this finding, we observed that T antigen suppressed the death program engaged by Bin1. This observation was interesting in light of emerging evidence that T antigen has roles in cell immortalization and human cell transformation beyond Rb and p53 inactivation. In support of a link to c-Myc-induced death processes, AEBSF, a serine protease inhibitor that inhibits apoptosis by c-Myc, potently suppressed DNA degradation by Bin1. Our findings suggest that the tumor suppressor activity of Bin1 reflects engagement of a unique cell death program. We propose that loss of Bin1 may promote malignancy by blunting death penalties associated with oncogene activation. PMID- 11032018 TI - Rit, a non-lipid-modified Ras-related protein, transforms NIH3T3 cells without activating the ERK, JNK, p38 MAPK or PI3K/Akt pathways. AB - The biological functions of Rit (Ras-like protein in tissues) and Rin (Ras-like protein in neurons), members of a novel branch of Ras-related GTP-binding proteins that are approximately 50% identical to Ras, have not been characterized. Therefore, we assessed their activity in growth control, transformation and signaling. NIH cells stably expressing a constitutively activated mutant of Rit [Rit(79L)] (analogous to the oncogenic mutant H-Ras(61L)) demonstrated strong growth transformation, proliferating rapidly in low serum and forming colonies in soft agar and tumors in nude mice. Although Rit(79L) alone did not promote morphologically transformed foci, it cooperated with both Raf and Rho A to form Rac/Rho-like foci. Rin [Rin(78L)] cooperated only with Raf. Rit(79L) but not Rin(78L) stimulated transcription from luciferase reporter constructs regulated by SRF, NF-kappaB, Elk-1 and Jun. However, neither activated ERK, JNK or p38, or PI3-K/Akt kinases in immune complex kinase assays. Interestingly, although Rit lacks any known recognition signal for C-terminal lipidation, Rit-transformed cell growth and survival in low serum is dependent on a farnesylated protein, as treatment with farnesyltransferase inhibitors caused apoptosis. Rin cooperated with Raf in focus assays but did not otherwise function in these assays, perhaps due to a lack of appropriate effector pathways in NIH3T3 fibroblasts for this neural-specific Ras family member. In summary, although Rit shares most core effector domain residues with Ras, our results suggest that Rit uses novel effector pathways to regulate proliferation and transformation. PMID- 11032020 TI - Identification and characterization of JunD missense mutants that lack menin binding. AB - Menin, the product of the MEN1 tumor suppressor gene, binds to the AP1 transcription factor JunD and represses JunD transcriptional activity. The effects of human or mouse JunD missense mutations upon menin interaction were studied by random and alanine scanning mutagenesis of the menin binding region of JunD (amino acids 1-70). JunD mutant proteins were tested for menin binding in a reverse yeast two-hybrid assay, and for transcriptional regulation by menin in AP1-reporter assays. Random mutagenesis identified two different mutations that disrupted menin interaction at mouse JunD amino acid 42 (G42E and G42R). Mutation G42A generated by alanine scanning did not affect menin binding, likely reflecting the conserved nature of this amino acid substitution. Furthermore, by size exclusion chromatography menin co-migrated with wild type JunD but not with the JunD mutant tested (G42E). Alanine scanning mutagenesis of residues 30-55 revealed two different amino acids, P41 and P44, of mouse JunD that were critical for interaction with menin. Mouse JunD missense mutants P41A, G42R, G42E and P44A failed to bind menin and also escaped menin's control over their transcriptional activity. At lower amounts of transfected menin, the transcriptional effect of menin on the mutants P41A, G42R and G42E was changed from repression to activation, similar to that with c-jun. In conclusion, a small N-terminal region of JunD mediates a key difference between JunD and c-jun, and a component of this difference is dependent on JunD binding to menin. PMID- 11032019 TI - Human prolactin (hPRL) antagonists inhibit hPRL-activated signaling pathways involved in breast cancer cell proliferation. AB - The involvement of human prolactin (hPRL) in breast cancer has been recently reconsidered based on its autocrine/paracrine proliferative effect described in human mammary tumor epithelial cells. Therefore, there is growing interest in the development of potent hPRL antagonists that may inhibit this effect. We previously designed hPRL analogs displaying antagonistic properties in a human transcriptional bioassay. We now report that the most potent of those analogs, G129R-hPRL, antagonizes all hPRL-induced effects analysed in various breast cancer cell lines, including cell proliferation. The analog per se lacks intrinsic agonistic activity on PRL receptor-activated signaling cascades, cell proliferation and apoptosis, indicating that its mode of action only occurs through competitive inhibition of hPRL. We provide some molecular basis of this antagonistic effect by demonstrating that G129R-hPRL competitively inhibits hPRL activation of the JAK-STAT and MAPK pathways, two signaling cascades involved in the mitogenic effect of hPRL in mammary epithelial cells. This competitive inhibition persists for at least 48 h, as evidenced by long term analysis of STAT5b activation or of progression through cell cycle. These results are the first demonstration at the molecular level that hPRL antagonists interfering with receptor dimerization disrupt signaling events in breast cancer cells, which prevents hPRL-induced cell proliferation. PMID- 11032021 TI - Caspase-dependent apoptosis by ectopic expression of E2F-4. AB - E2F is a family of transcription factors which regulates cell cycle and apoptosis of mammalian cells. E2F-1-3 localize in the nucleus, and preferentially bind pRb, while E2F-4 and 5 have no nuclear localization signal and preferentially bind p107/p130. E2F-6 suppresses the transcriptional activity of other E2F proteins. DP-1 and 2 are heterodimeric partners of each E2F protein. Using tetracycline responsive promoters, here we compared the effects of ectopic expression of E2F 1, DP-1 and E2F-4 on cell cycle progression and apoptosis in Chinese hamster cell lines. We found that E2F-4, as well as DP-1 and E2F-1, induced growth arrest and caspase-dependent apoptosis. E2F-4 did not have a marked effect on cell cycle progression, while E2F-1 induced DNA synthesis of resting cells and DP-1 arrested cells in G1. Ectopic expression of E2F-4 did not activate E2F-dependent transcription. Our results suggest that expression of E2F-4 at elevated levels induces growth arrest and apoptosis of mammalian cells through a mechanism distinct from E2F-1 and DP-1. PMID- 11032023 TI - Kinetic analysis of Sp1-mediated transcriptional activation of the human DNA polymerase beta promoter. AB - In the present studies, we have examined the effect of Sp1 on the activation of the human DNA polymerase beta (beta-pol), a TATA-less promoter. A HeLa cell nuclear extract (NE) based in vitro runoff transcription system of core beta-pol promoter human DNA (pbetaP8) three-step kinetic model of transcription initiation were used to describe the kinetic effect of Sp1. The results showed that distal Sp1-binding sites in the core beta-pol promoter are important for transcriptional activation of the pbetaP8 promoter. A detailed kinetic analysis showed that Sp1 stimulates the activity of the pbetaP8 promoter through distal Sp1-binding sites by increasing the amount of recruitment, instead of stimulating the apparent rate of RPc assembly (k1). There was no significant effect of Sp1 on the apparent rate of open complex (RPo) formation (k2) or on the apparent rate of promoter clearance (k3) of the pbetaP8 promoter. These studies define the kinetic mechanisms by which Sp1 may regulate the rate of transcript formation of the pbetaP8 promoter, and these results may have implications for Sp1 regulation of TATA-less promoters. PMID- 11032022 TI - Genomic structure, chromosomal localization and identification of mutations in the xeroderma pigmentosum variant (XPV) gene. AB - The xeroderma pigmentosum variant (XP-V) is one of the most common forms of this cancer-prone syndrome. XP groups A through G are characterized by defective nucleotide excision repair, whereas the XP-V phenotype is proficient in this pathway. The XPV gene encodes DNA polymerase eta, which catalyzes an accurate translesion synthesis, indicating that the XPV gene contributes tumor suppression in normal individuals. Here we describe the genomic structure and chromosomal localization of the XPV gene, which includes 11 exons covering the entire coding sequence, lacks a TATA sequence in the upstream region of the transcription initiation, and is located at the chromosome band 6p21.1-6p12. Analyses of patient-derived XP-V cell lines strongly suggested that three of four cell lines carried homozygous mutations in the XPV gene. The fourth cell line, XP1RO, carried heterozygous point mutations in the XPV gene, one of which was located at the splice acceptor site of exon 2, resulting in the omission of exon 2 from the mature mRNA. These findings provide a basis for diagnosis and therapy of XP-V patients. PMID- 11032024 TI - Characterization of a novel quiescence responsive element downregulated by v-Src in the promoter of the neuroretina specific QR1 gene. AB - The neuroretina is a functional unit of the central nervous system which arises through successive steps of division, growth arrest and differentiation of neuroectodermal precursors. Postmitotic quail neuroretina (QNR) cells are conditionally induced to divide upon infection with temperature sensitive mutants of Rous sarcoma virus (RSV), since QNR cell division can be arrested by either inactivating p60v-Src at the nonpermissive temperature (41 degrees C) or by serum deprivation at 37 degrees C. We are studying the transcriptional control of QR1, a neuroretina specific gene, whose expression is down-regulated in proliferating cells at 37 degrees C and is fully restored when these cells are made quiescent. We previously showed that this quiescence specific upregulation implicates a promoter region named A box, which binds Maf transcription factors. We report the identification of the C box, a second promoter sequence that activates QR1 transcription in non dividing cells. This sequence is able to form two DNA protein complexes, one of which (C4) is predominantly detected in growth arrested NR cells. We identified the DNA binding site for C4 and described mutations that abolish both C4 binding and promoter activity in quiescent cells. Moreover, we show that a multimerized C box is able to stimulate a heterologous promoter in non dividing cells and constitutes, therefore, a novel quiescence responsive enhancer. Finally, we report that QR1 transcriptional response to cell quiescence requires cooperation between the C box and A box. PMID- 11032025 TI - Reversal of an antiestrogen-mediated cell cycle arrest of MCF-7 cells by viral tumor antigens requires the retinoblastoma protein-binding domain. AB - Proliferation of MCF-7 cells is estrogen dependent and antiestrogen sensitive. In the absence of estrogens or presence of antiestrogens MCF-7 cells arrest in the G1 phase of the cell cycle, and this arrest is associated with an accumulation of the active, hypophosphorylated form of the retinoblastoma protein (pRb). Because active pRb negatively regulates passage from G1 to S phase, this suggests that pRb is a crucial target of estrogen action, and that its inactivation might lead to antiestrogen resistance. We tested this hypothesis by expressing viral tumor antigens (T antigens), which bind and inactivate pRb, in MCF-7 cells, and determining the effects on cell proliferation in the presence of antiestrogens. The results of these experiments demonstrate that T antigen expression confers antiestrogen resistance to MCF-7 cells. Using a panel of mutant T antigens, we further demonstrate that the pRb-binding, but not the p53 binding domain is required to confer antiestrogen resistance. Thus, pRb is an important target of estrogen action, and its inactivation can contribute to the development of antiestrogen resistance. PMID- 11032026 TI - Downmodulation of caveolin-1 expression in human ovarian carcinoma is directly related to alpha-folate receptor overexpression. AB - Caveolin (cav-1) and the GPI-anchored alpha-folate receptor (alphaFR) are membrane proteins both found associated to caveolar structures. Several studies in tumor cells independently reported cav-1 downregulation and alphaFR overexpression. Here we analysed the expression of the two molecules in normal and tumor ovarian samples derived from fresh specimens and from cultured cell lines. Whereas normal ovary surface epithelial cells displayed only cav-1 expression, ovarian tumor surgical samples and cell lines (COR, IGROV1, OVCAR3 and OVCA432) displayed high alphaFR and low-level or no cav-1 expression, except those cell lines (SKOV3 and SW626) with the lower alphaFR expression. SKOV3, but not two alphaFR-negative non-ovarian cell lines, exhibited down-regulation of cav 1 expression following stable alphaFR cDNA transfection. Conversely, cav-1 transfection in IGROV1 cells led to downregulated alphaFR expression, together with formation of caveolar structures and reduction of growth capability. Moreover, cav-1 expression was induced in IGROV1 cells by transfection with intracellular anti-alphaFR antibodies to downmodulate alphaFR expression. In cav 1 transfected cells, transcriptional activity of the alphaFR-specific promoter P1 was reduced by 70% and an additional specific DNA-protein complex was identified by gel-shift assay, indicating that cav-1 expression influences alphaFR gene transcription. Together these results support the notion that alphaFR and cav-1 protein expression is reciprocally regulated in ovary cancer cells. PMID- 11032027 TI - Differential regulation of human RecQ family helicases in cell transformation and cell cycle. AB - Three human RecQ DNA helicases, WRN, BLM and RTS, are involved in the genetic disorders associated with genomic instability and a high incidence of cancer. RecQL1 and RecQL5 also belong to the human RecQ helicase family, but their correlation with genetic disorders, if any, is unknown. We report here that in human B cells transformed by Epstein-Barr virus (EBV), human fibroblasts and umbilical endothelial cells transformed by simian virus 40, the expression of WRN, BLM, RTS and RecQL1 was sharply up-regulated. In B cells this expression was stimulated within 5-40 h by the tumor promoting agent phorbol myristic acetate (PMA). Interestingly, RecQL5beta, an alternative splicing product of RecQL5 with a nuclear localization signal, is expressed in resting B cells without significant modulation of its synthesis by EBV or PMA, suggesting it has a role in resting cells. We also roughly determined the number of copies per cell for the five RecQ helicase in B cells. In addition, levels of the different RecQ helicases are modulated in different ways during the cell cycle of actively proliferating fibroblasts and umbilical endothelial cells. Our results support the view that the levels of WRN, BLM, RTS and RecQL1 are differentially up regulated to guarantee genomic stability in cells that are transformed or actively proliferating. PMID- 11032028 TI - Identification of an RNA element that confers post-transcriptional repression of connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene: similarities to retroviral RNA-protein interactions. AB - The repressive effect of the 3'-untranslated region (3'-UTR) in human connective tissue growth factor/ hypertrophic chondrocyte specific 24 (ctgf/hcs24) mRNA on gene expression had been demonstrated in our previous study. Here, we identified a minimal RNA element in the 3'-UTR, which acts as a cis-acting element of structure-anchored repression (CAESAR). Deletion analyses of the 3'-UTR led us to minimize the element of 84 bases at the junction of the coding region and the 3' UTR. The minimized RNA segment is predicted, and actually capable of forming a stable secondary structure in vitro. Mutational analyses disclosed a significant relationship between the predicted structure and repressive effect. The utility of CAESAR as a post-transcriptional regulatory element was represented by the fact that steady-state mRNA levels were not affected by CAESAR linked in cis, while protein levels from such a chimeric gene were markedly reduced. Of note, the CAESAR sequence exerted no effect, when it was placed upstream of the promoter. Finally, RNA gel electromobility-shift analyses demonstrated a nuclear factor that interacts with the folded CAESAR. Taken together, it was uncovered that CAESAR of ctgf is a novel post-transcriptional structured RNA regulatory element, probably acting through direct interactions with a nuclear factor as observed in retroviral RNA elements with certain proteins. PMID- 11032029 TI - Additive effect between NF-kappaB subunits and p53 protein for transcriptional activation of human p53 promoter. AB - The tumor suppressor p53 plays a pivotal role in the cellular response to DNA damage as it controls DNA repair, cell cycle arrest and apoptosis. We studied the autoregulation of human p53 gene transcription in colon cancer cell lines. Wild type p53 has been shown to autoregulate its own transcription either positively or negatively and probably in a cell-type-specific manner. Indeed, a p53 binding site has been described in the human and murine p53 promoters, but a direct binding of wild-type p53 protein to this site has never been reported. In this study, we demonstrated a transactivation of human p53 promoter by wild-type p53 in human colon cancer cells. We identified in the human p53 promoter a novel potential p53-responsive element that binds wild-type p53. Moreover, wild-type p53 protein transactivated a reporter plasmid containing a luciferase gene driven by a minimal promoter harboring this p53 binding site. Finally, as the p53 promoter contains an NF-kappaB binding site, we demonstrated an additive effect when NF-kappaB subunits and p53 protein combined to transactivate the human p53 promoter. PMID- 11032030 TI - The JAK-inhibitor, JAB/SOCS-1 selectively inhibits cytokine-induced, but not v Src induced JAK-STAT activation. AB - Recently, constitutive activation of JAK kinases (JAKs) and/or signal transducers and activators of transcription (STATs) has been reported in growing numbers of human cancer cells as well as oncogene-transformed cells. JAB/SOCS-1 has been shown to be an intrinsic JAK tyrosine kinase inhibitor and to suppress the cytokine-dependent JAK-STAT pathway. In this report, we investigated the effect of ectopic expression of JAB on v-Src-induced JAK-STAT activation. Forced expression of JAB in v-Src-transformed NIH3T3 cells neither suppressed phosphorylation of STAT3 and JAK1/JAK2 nor blocked STAT3-reporter gene activation. Colony forming assay also showed that JAB did not suppress v-Src induced transformation of NIH3T3 cells, while dominant negative STAT3 suppressed it. In contrast, JAB could downregulate phosphorylation of STAT1 and STAT3 induced by interferon gamma (IFNgamma) and interleukin-6 (IL-6) plus soluble IL6 receptor (sIL-6R), respectively. Furthermore, in vitro kinase assay indicated that JAB suppressed hyperactivation of JAK1/JAK2 and JAK1 induced by IFNgamma and IL-6 plus sIL-6R respectively, but not v-Src-induced basal JAK1/JAK2 activity. Nevertheless, both JAK1/JAK2 activated by v-Src and that activated by IL-6 plus sIL-6R could similarly bind JAB. These results clearly demonstrate that JAB distinguishes cytokine-induced JAK-STAT signaling from v-Src-induced one and can not suppress the transformation with v-Src. PMID- 11032032 TI - Spike-driven synaptic plasticity: theory, simulation, VLSI implementation. AB - We present a model for spike-driven dynamics of a plastic synapse, suited for aVLSI implementation. The synaptic device behaves as a capacitor on short timescales and preserves the memory of two stable states (efficacies) on long timescales. The transitions (LTP/LTD) are stochastic because both the number and the distribution of neural spikes in any finite (stimulation) interval fluctuate, even at fixed pre- and postsynaptic spike rates. The dynamics of the single synapse is studied analytically by extending the solution to a classic problem in queuing theory (Takacs process). The model of the synapse is implemented in aVLSI and consists of only 18 transistors. It is also directly simulated. The simulations indicate that LTP/LTD probabilities versus rates are robust to fluctuations of the electronic parameters in a wide range of rates. The solutions for these probabilities are in very good agreement with both the simulations and measurements. Moreover, the probabilities are readily manipulable by variations of the chip's parameters, even in ranges where they are very small. The tests of the electronic device cover the range from spontaneous activity (3-4 Hz) to stimulus-driven rates (50 Hz). Low transition probabilities can be maintained in all ranges, even though the intrinsic time constants of the device are short (approximately 100 ms). Synaptic transitions are triggered by elevated presynaptic rates: for low presynaptic rates, there are essentially no transitions. The synaptic device can preserve its memory for years in the absence of stimulation. Stochasticity of learning is a result of the variability of interspike intervals; noise is a feature of the distributed dynamics of the network. The fact that the synapse is binary on long timescales solves the stability problem of synaptic efficacies in the absence of stimulation. Yet stochastic learning theory ensures that it does not affect the collective behavior of the network, if the transition probabilities are low and LTP is balanced against LTD. PMID- 11032031 TI - Characterization of a novel ETS gene, TELB, encoding a protein structurally and functionally related to TEL. AB - The TEL/ETV6 gene is located at 12p13 and is frequently involved in chromosomal translocations in human malignancies usually resulting in the expression of fusion proteins between the amino terminal part of TEL, and either unrelated transcription factors or protein tyrosine kinases. We report here a novel gene named TELB which is located on human chromosomal band 6p21 and encodes a protein highly related to TEL. TELB is widely expressed in different tissues and, similarly to TEL encodes a sequence-specific transcriptional repressor. PMID- 11032034 TI - Representation of concept lattices by bidirectional associative memories. AB - This article presents a concept interpretation of patterns for bidirectional associative memory (BAM) and a representation of hierarchical structures of concepts (concept lattices) by BAMs. The constructive representation theorem provides a storing rule for a training set that allows a concept interpretation. Examples demonstrating the theorems are presented. PMID- 11032033 TI - Modeling alternation to synchrony with inhibitory coupling: a neuromorphic VLSI approach. AB - We developed an analog very large-scale integrated system of two mutually inhibitory silicon neurons that display several different stable oscillations. For example, oscillations can be synchronous with weak inhibitory coupling and alternating with relatively strong inhibitory coupling. All oscillations observed experimentally were predicted by bifurcation analysis of a corresponding mathematical model. The synchronous oscillations do not require special synaptic properties and are apparently robust enough to survive the variability and constraints inherent in this physical system. In biological experiments with oscillatory neuronal networks, blockade of inhibitory synaptic coupling can sometimes lead to synchronous oscillations. An example of this phenomenon is the transition from alternating to synchronous bursting in the swimming central pattern generator of lamprey when synaptic inhibition is blocked by strychnine. Our results suggest a simple explanation for the observed oscillatory transitions in the lamprey central pattern generator network: that inhibitory connectivity alone is sufficient to produce the observed transition. PMID- 11032036 TI - Efficient event-driven simulation of large networks of spiking neurons and dynamical synapses. AB - A simulation procedure is described for making feasible large-scale simulations of recurrent neural networks of spiking neurons and plastic synapses. The procedure is applicable if the dynamic variables of both neurons and synapses evolve deterministically between any two successive spikes. Spikes introduce jumps in these variables, and since spike trains are typically noisy, spikes introduce stochasticity into both dynamics. Since all events in the simulation are guided by the arrival of spikes, at neurons or synapses, we name this procedure event-driven. The procedure is described in detail, and its logic and performance are compared with conventional (synchronous) simulations. The main impact of the new approach is a drastic reduction of the computational load incurred upon introduction of dynamic synaptic efficacies, which vary organically as a function of the activities of the pre- and postsynaptic neurons. In fact, the computational load per neuron in the presence of the synaptic dynamics grows linearly with the number of neurons and is only about 6% more than the load with fixed synapses. Even the latter is handled quite efficiently by the algorithm. We illustrate the operation of the algorithm in a specific case with integrate-and fire neurons and specific spike-driven synaptic dynamics. Both dynamical elements have been found to be naturally implementable in VLSI. This network is simulated to show the effects on the synaptic structure of the presentation of stimuli, as well as the stability of the generated matrix to the neural activity it induces. PMID- 11032035 TI - A silicon implementation of the fly's optomotor control system. AB - Flies are capable of stabilizing their body during free flight by using visual motion information to estimate self-rotation. We have built a hardware model of this optomotor control system in a standard CMOS VLSI process. The result is a small, low-power chip that receives input directly from the real world through on board photoreceptors and generates motor commands in real time. The chip was tested under closed-loop conditions typically used for insect studies. The silicon system exhibited stable control sufficiently analogous to the biological system to allow for quantitative comparisons. PMID- 11032037 TI - Clustering irregular shapes using high-order neurons. AB - This article introduces a method for clustering irregularly shaped data arrangements using high-order neurons. Complex analytical shapes are modeled by replacing the classic synaptic weight of the neuron by high-order tensors in homogeneous coordinates. In the first- and second-order cases, this neuron corresponds to a classic neuron and to an ellipsoidalmetric neuron. We show how high-order shapes can be formulated to follow the maximum-correlation activation principle and permit simple local Hebbian learning. We also demonstrate decomposition of spatial arrangements of data clusters, including very close and partially overlapping clusters, which are difficult to distinguish using classic neurons. Superior results are obtained for the Iris data. PMID- 11032038 TI - Learning chaotic attractors by neural networks. AB - An algorithm is introduced that trains a neural network to identify chaotic dynamics from a single measured time series. During training, the algorithm learns to short-term predict the time series. At the same time a criterion, developed by Diks, van Zwet, Takens, and de Goede (1996) is monitored that tests the hypothesis that the reconstructed attractors of model-generated and measured data are the same. Training is stopped when the prediction error is low and the model passes this test. Two other features of the algorithm are (1) the way the state of the system, consisting of delays from the time series, has its dimension reduced by weighted principal component analysis data reduction, and (2) the user adjustable prediction horizon obtained by "error propagation"-partially propagating prediction errors to the next time step. The algorithm is first applied to data from an experimental-driven chaotic pendulum, of which two of the three state variables are known. This is a comprehensive example that shows how well the Diks test can distinguish between slightly different attractors. Second, the algorithm is applied to the same problem, but now one of the two known state variables is ignored. Finally, we present a model for the laser data from the Santa Fe time-series competition (set A). It is the first model for these data that is not only useful for short-term predictions but also generates time series with similar chaotic characteristics as the measured data. PMID- 11032039 TI - Generalized discriminant analysis using a kernel approach. AB - We present a new method that we call generalized discriminant analysis (GDA) to deal with nonlinear discriminant analysis using kernel function operator. The underlying theory is close to the support vector machines (SVM) insofar as the GDA method provides a mapping of the input vectors into high-dimensional feature space. In the transformed space, linear properties make it easy to extend and generalize the classical linear discriminant analysis (LDA) to nonlinear discriminant analysis. The formulation is expressed as an eigenvalue problem resolution. Using a different kernel, one can cover a wide class of nonlinearities. For both simulated data and alternate kernels, we give classification results, as well as the shape of the decision function. The results are confirmed using real data to perform seed classification. PMID- 11032040 TI - Generalization and selection of examples in feedforward neural networks. AB - In this work, we study how the selection of examples affects the learning procedure in a boolean neural network and its relationship with the complexity of the function under study and its architecture. We analyze the generalization capacity for different target functions with particular architectures through an analytical calculation of the minimum number of examples needed to obtain full generalization (i.e., zero generalization error). The analysis of the training sets associated with such parameter leads us to propose a general architecture independent criterion for selection of training examples. The criterion was checked through numerical simulations for various particular target functions with particular architectures, as well as for random target functions in a nonoverlapping receptive field perceptron. In all cases, the selection sampling criterion lead to an improvement in the generalization capacity compared with a pure random sampling. We also show that for the parity problem, one of the most used problems for testing learning algorithms, only the use of the whole set of examples ensures global learning in a depth two architecture. We show that this difficulty can be overcome by considering a tree-structured network of depth 2log2(N)-1. PMID- 11032041 TI - Stationary and integrated autoregressive neural network processes. AB - We consider autoregressive neural network (AR-NN) processes driven by additive noise and demonstrate that the characteristic roots of the shortcuts-the standard conditions from linear time-series analysis-determine the stochastic behavior of the overall AR-NN process. If all the characteristic roots are outside the unit circle, then the process is ergodic and stationary. If at least one characteristic root lies inside the unit circle, then the process is transient. AR-NN processes with characteristic roots lying on the unit circle exhibit either ergodic, random walk, or transient behavior. We also analyze the class of integrated AR-NN (ARI-NN) processes and show that a standardized ARI-NN process "converges" to a Wiener process. Finally, least-squares estimation (training) of the stationary models and testing for nonstationarity is discussed. The estimators are shown to be consistent, and expressions on the limiting distributions are given. PMID- 11032042 TI - Learning to forget: continual prediction with LSTM. AB - Long short-term memory (LSTM; Hochreiter & Schmidhuber, 1997) can solve numerous tasks not solvable by previous learning algorithms for recurrent neural networks (RNNs). We identify a weakness of LSTM networks processing continual input streams that are not a priori segmented into subsequences with explicitly marked ends at which the network's internal state could be reset. Without resets, the state may grow indefinitely and eventually cause the network to break down. Our remedy is a novel, adaptive "forget gate" that enables an LSTM cell to learn to reset itself at appropriate times, thus releasing internal resources. We review illustrative benchmark problems on which standard LSTM outperforms other RNN algorithms. All algorithms (including LSTM) fail to solve continual versions of these problems. LSTM with forget gates, however, easily solves them, and in an elegant way. PMID- 11032043 TI - Alexander V. Koranyi, who first coined the term "Renal Insufficiency" and characterized it functionally. PMID- 11032044 TI - Apoptosis induction and inhibition of cellular proliferation by angiotensin II: possible implication and perspectives. AB - The renin-angiotensin system plays a pivotal role in the regulation of fluid, electrolyte metabolism and blood pressure. Molecular cloning and pharmacological studies have defined two major classes of Angiotensin II (Ang II) receptors, designated AT1 and AT2. Recently, it has been well recognized that Ang II, beside its classical physiological actions, is a profibrogenic peptide and displays characteristics of a growth factor. The emerging picture suggests that angiotensin receptor subtypes exert opposing features in many aspects of their biological function, most importantly in cellular growth and proliferation. Accordingly, the proliferative and/or growth-promoting effects of Ang II are thought to be mediated by AT1 receptor, whereas the AT2 receptor subtype may have growth-inhibitory properties. The novel finding that Ang II is able to induce apoptosis by AT2 receptors in diverse cell types is of great scientific interest, as recent studies revealed a role for apoptosis as a deliberate form of cell death in the pathogenesis of various cardiovascular diseases such as heart failure and vascular remodeling. Furthermore apoptotic cell death might occur during the development of progressive glomerulosclerosis. It is tempting to speculate that autocrine-paracrine vasoactive substances such as Ang II might regulate these apoptotic processes during pathogenic conditions. PMID- 11032045 TI - Influence of a single treatment with vitamin E or K (hormonal imprinting) of neonatal rats on the sexual behavior of adults. AB - The effect of a single neonatal treatment (imprinting) with vitamin E or vitamin K1 on the sexual activity of three-month old rats, was studied. In female animals vitamin E treatment significantly lowered the Meyerson index and lordosis quotient, among males there were significantly more inactive animals and no multiple ejaculations could be observed. Vitamin K1 treatment caused only slight changes in the same direction, in both sexes. Considering also earlier results concerning vitamin A and D neonatal treatments (alterations in receptor binding capacity, sex hormone levels and sexual behavior), and receptorial changes caused by neonatal vitamin E and K1 treatments, the present experiment also calls attention to the lifelong effects of perinatal treatment with lipid soluble vitamins. PMID- 11032046 TI - Isoluminant monochromatic and polychromatic visual backgrounds alter response characteristics of cat visual neurons after stimulation with stationary and moving light bars. AB - The role of colour vision in night-active cats has not been elucidated completely hitherto. In order to assess the colour sensitivity in cat cortical neurons we used large isoluminant computer-generated monochromatic and polychromatic background stimuli which were superimposed on moving and stationary (on/off) light bars. Background stimuli were moved at different speeds either inphase or antiphase. The modulatory effect of the visual noise on the neuronal bar was the primary objective of the study. The maximum PSTH peaks of some 40% of the neurons tested was influenced by both moving and stationary bars. About 2 thirds of maximum peak-sensitive cells showed also altered direction selectivity. Latencies and field widths, on the other hand, turned out to be rather stable. The retino cortical conduction time was not influenced either. In conclusion, a large portion of cat cortical visual neurons is remarkably sensitive to the spectral composition of the visual noise process surrounding the stimulating light bar. PMID- 11032047 TI - Acute 4-aminopyridine seizures increase the regional cerebral blood flow in the thalamus and neocortex, but not in the entire allocortex of the mouse brain. AB - Systemic injections of 4-aminopyridine precipitate epileptiform generalized seizures characterized mainly by shivering of the body, tail movements and tonic clonic convulsions in rats and mice. However, only few details are known as concerns which brain regions are possibly affected and stimulated by the compound. The aim of the present study was to investigate the changes in regional cerebral blood flow in mice by using the lipophilic compound technetium-99m hexamethyl-propyleneamineoxime (99mTc-HMPAO). Whilst the uptake of 99mTc-HMPAO was increased significantly in the neocortex and thalamus following the induction of acute 4-aminopyridine seizures, no such changes were observed in the allocortex of the mice. The increases in uptake in the neocortex and thalamus were completely prevented by carbamazepine (which abolished the symptoms of the seizure, too). The primary involvement of the neocortex and thalamus points to the importance of thalamocortical circuits in the precipitation and maintenance of experimental 4-aminopyridine convulsions. PMID- 11032048 TI - Synergistic action of rutin and orthovanadate on nitric oxide release from mouse macrophage cells. AB - The cooperative action of sodium orthovanadate (a putative protein phosphotyrosine phosphatase inhibitor) and rutin (an effective superoxide scavenger) on the nitric oxide (NO) production of J774A.1 mouse macrophage cells has been investigated. Orthovanadate alone caused a mild but significant increase in NO production of the cells at its highest concentration used (500 microM). Orthovanadate and rutin together caused a significant increase in the nitrite level of the supematants of the J774A.1 cells after a 24-hour incubation period, in a concentration dependent manner. The optimal doses for orthovanadate and rutin were 50 microM and 100 microM, respectively. This cooperative action of rutin and orthovanadate was totally inhibitable by catalase, reduced glutathion, N-acetylcystein, cycloheximide, pyrrolidine dithiocarbamate (a putative NF-kappaB inhibitor), genistein and tyrphostin-AG126 (two protein tyrosine-kinase inhibitors). Superoxide dismutase had no inhibitory effect. Orthovanadate and rutin (only together) could induce the oxidation of 2',7'-dichlorofluorescein diacetate, a marker of hydrogen peroxide. This effect was inhibitable by reduced glutathion, a hydrogen peroxide specific scavenger. These findings suggest, that orthovanadate can induce the production of NO by J774A.1 macrophages not only by inhibition of protein tyrosine-phosphatases, but, using it with rutin, by increasing the level of hydrogen peroxide in the cells. PMID- 11032049 TI - Risk assessment of neurotoxic pesticides. AB - In order to establish safe exposure levels for toxic chemicals, risk assessment guidelines have been developed. A compilation is given by the author on the elements of risk assessment of hazardous neurotoxic pesticides, using data obtained from human epidemiological studies, from animal experiments, from the international literature and from the author's own experiments as well. Well controlled laboratory studies of neurotoxicity have the potential to provide adequate exposure and effect data for accurate hazard identification. Animal models of neurotoxicity as highly sensitive behavioral and neurophysiological methods as a function of doses, provide data for human low dose extrapolation by using mathematical models. This procedure might be the basis for reducing risk ("risk management"), therefore some examples are given, how to handle properly neurotoxic pesticides with different- high or low-risk. PMID- 11032050 TI - A chloroform extract obtained from a decoction of Ficus carica leaves improves the cholesterolaemic status of rats with streptozotocin-induced diabetes. AB - The antidiabetic effects of Ficus carica leaf extracts have been reported previously. From the aqueous decoction of fig leaves, after treatment with HCI, centrifuging, treatment with sodium hydroxide (NaOH) and extraction with chloroform (CHCl3), the administration of the organic phase rats with streptozotocin-induced diabetes led to a decline in the levels of total cholesterol and an decrease in the total cholesterol/HDL cholesterol ratio (with respect to the control group), together with a reduction of the hyperglycaemia. PMID- 11032052 TI - Use of isoptin for counteracting the effect of methol on cardiac activity in experimental animals. AB - The purpose of the present study is to use isoptin in order to lower intracellular Ca2+ concentration for counteracting the effect of methol on cardiac activity in acute intoxication of the experimental animals. Adult male Wistar rats were used - 8 animals/lot anaesthesized with urethan. ECG activity was recorded every 15 minutes, for 360 minutes. Administration (oral) of methol in a dose of 310 mg/kg/bw. (DL100) produced sinusal bradycardia, which became more marked later on. Isoptin administration in a dose of 5 mg/kg/bw. in rats with 310 mg/kg/bw. methol decreased the heart rate, the lowest value occurring 120 minutes after administration, followed by a slow increase until the end of the recording. When isoptin was given in a dose of 2.5 mg/kg/bw. to rats with 310 mg/kg/bw. methol, the decrease of the heart rate was much slower than before, for a period of 255 minutes, followed by a slight increase until the end of the recording. Isoptin counteracts the bradycardia - producing effect of methol in rats during the study time (6 hours), and the effect after this period of time requires further investigations. PMID- 11032051 TI - The effects of the initial arterial tone on the pressure responses to phenylephrine. AB - The effect of the elevated arterial tone on pressure responses to stimulation of arterial alpha-adrenoreceptors by phenylephrine hydrochloride was studied in anesthetized Wistar rats. Different levels of the arterial tone and, hence of the mean arterial pressure, were established by means of angiotensin II infusion in the range from 101 to 160 mmHg. An elevation of the arterial tone led to a significant reduction of the arterial pressure and peripheral resistance rise produced by phenylephrine. The degree of relative reduction of the increase in the diastolic pressure exceeded 1.3 times that in the systolic pressure. The shifts of cardiac outputs remained unchanged. After cessation of angiotensin II infusion the restoration of the arterial pressure took place almost till the initial level. At this time the pressure effects of phenylephrine were tended to recovery. It is suggested that the elevated arterial tone attenuates the systemic pressure response to stimulation of arterial alpha-adrenoreceptors by a vascular mechanism based on a transmural pressure changes evoked by the constriction of the arterial vessels. PMID- 11032053 TI - The influence of intracerebroventricular administration of (+/-) propranolol and (+/-) verapamil on experimental myocardial ischemia and necrosis in rats. AB - In albino rats, infarctoid myocardial lesions were produced by intraperitoneal (i.p.) administration of isoproterenol (75 mg/kg, during 3 days). In other groups, the descending anterior left coronary artery was ligated. In both experimental settings, the intracerebroventricular (i.c.v.) administration of (+/ ) propranolol (100-200-300 microg/animal/day, during 7 days) or (+/-) verapamil (40-80-160 microg/animal/day, during 7 days) afforded a significant protection (with the exception of the lowest dose) on the investigated parameters: arrhythmias, ischemic zone (in coronary ligated rats), lactate dehydrogenase and aspartate aminotransferase activity of the serum, focal necrosis (in isoproterenol treated rats). This protective activity is lower than that afforded by i.p. administered (+/-) propranolol (5 mg/kg, during seven days) or (+/-) verapamil (5 mg/kg, during seven days). From these data it may be concluded that (+/-) propranolol and (+/-) verapamil have a protective action on the experimental myocardial ischemia and necrosis in rats, not only when the drugs come in direct contact with the heart, but also acting upon the central nervous system. PMID- 11032054 TI - Effects of ICI 118.551, a selective beta-2 adrenergic blocking agent on the guinea pig cardiac excitability and ventricular fibrillation threshold. AB - In isolated guinea pig perfused hearts ICI 118.551, a selective beta 2 adrenoceptor antagonist, induced transient ventricular extrasystoles. Following the termination of the perfusion, a very significant increase of both the ventricular fibrillation threshold and the refractory periods were measured. In guanethidine pretreated hearts, ICI 118.551 failed to induce premature beats. At the same time the fibrillation threshold and refractory periods exhibited a very significant decrease. The perfusion of equimolecular concentration of metoprolol, a beta-1-adrenoceptor antagonist, and (+) propranolol, a quinidine-like compound, induced, in most experimental settings, similar results as ICI 118.551. Thus, besides its beta-2-adrenoceptor antagonist properties, ICI 118.551 presented other pharmacological actions. PMID- 11032055 TI - Depigmentation of the normally pigmented patches in universal vitiligo patients by cryotherapy. AB - BACKGROUND: Complete depigmentation may occur, albeit rarely, in patients with universal vitiligo. They usually have one or more pigmented patches that remain normal, most frequently over the malar area of the face and dorsal aspects of forearms. Total repigmentation may not occur in these patients, even after receiving 150 session of PUVA therapy alone or in conjunction with other medical or surgical procedures, and there is possibility of recurrence after cessation of therapy. These patients are usually more interested in depigmentation rather than trying for repigmentation. OBJECTIVE: Because of the relatively high sensitivity of melanocytes to cryotherapy and the possibility of isomorphic phenomenon in vitiligo patients, removal of the remaining normally pigmented patches was attempted in patients with universal vitiligo using cryotherapy. METHOD: Five volunteer patients with universal vitiligo were chosen, all of whom wished to have their pigmented patches removed. In all five patients a small area was tested first in order to assure the patients of what the procedure involved and to show its likely result. All pigmented areas were then treated to 1-3 sessions of cryotherapy using a closed contact CO2 cryogun, with 4-6-week intervals. RESULTS: Complete and permanent depigmentation was achieved in all five patients with excellent cosmetic results and no complications or scarring. CONCLUSION: Cryotherapy is a cost effective, non-complicating, easily available procedure which can be used for depigmentation of normally pigmented patches in patients with universal vitiligo. PMID- 11032056 TI - Patient attitudes to topical antipsoriatic treatment with calcipotriol and dithranol. AB - OBJECTIVE: Assessment of patient preference for antipsoriatic treatment with calcipotriol ointment or short-contact dithranol cream. METHODS: Two hundred and fifty-eight psoriatic patients treated with calcipotriol (n = 138) or dithranol (n = 120) for up to 3 months, assessed the acceptability of treatment, overall satisfaction with treatment, their treatment preference using the 'willingness to pay' principle and selected their treatment of choice. RESULTS: Overall satisfaction with calcipotriol was significantly better (72.7%, dithranol 60.3%; odds ratio 1.75, 95% CI 1.03, 2.99: P = 0.04). Patients considered calcipotriol a more acceptable treatment than dithranol in its appearance, smell, non-irritancy, method and ease of application and lack of staining. Dithranol was considered less sticky than calcipotriol. Patients were 'willing to pay' a mean of pound sterling 12.16 monthly for calcipotriol and pound sterling 10.66 monthly for dithranol. 'Willingness to pay' did not correlate well with overall treatment satisfaction and was not correlated with household income. Calcipotriol was the preferred treatment of choice (calcipotriol 63%, dithranol 24%). CONCLUSION: Patients with psoriasis prefer treatment with calcipotriol ointment over short contact dithranol cream. PMID- 11032057 TI - Second intention healing after Mohs micrographic surgery. AB - BACKGROUND: Although second intention healing has been used for many years in Mohs micrographic surgery, it has lost popularity. This is due to the long process and functional alteration of important anatomical units if certain wound healing factors are not carefully taken into consideration. OBJECTIVE: To review indications, contraindications, complications, advantages, disadvantages, as well as basic concepts of the wound healing process, wound care and antibiotic prophylaxis in the management of open wounds after Mohs surgery. METHODS: A short but comprehensive review of studies published in the literature dealing with second intention healing as a reconstruction alternative option in certain situations. CONCLUSIONS: Second intention healing is a simple and cost-effective method for reconstruction after Mohs micrographic surgery in well-defined cases. It allows adequate tumoral control with good to excellent functional and cosmetic results. PMID- 11032058 TI - The role of HLA antigens in the development of paracoccidioidomycosis. AB - BACKGROUND: Paracoccidioidomycosis is a systemic granulomatous disease that involves primarily the lungs and may disseminate to other organs and systems. It is caused by Paracoccidioides brasiliensis, a fungus that exhibits reversible thermal dimorphism and whose natural habitat is presently unknown. There are two main clinical forms: the acute (subacute) juvenile form and the chronic adult form. The former runs a more rapid course and is more severe than the latter. This mycosis is found throughout Latin America. Brazil accounts for 80% of reported cases. Presumably P. brasiliensis thrives in humid and hot places, especially near forests or farms. The infection is endemic in certain areas, especially in Brazil, Colombia and Venezuela, where nearly 100% of the population show cutaneous paracoccidioidina positive skin tests, indicating previous contact with the fungus, although a small percentage show clinical manifestations of the disease. METHODS: We compared the expression of HLA class I antigens in a healthy group (control) and in a group of patients with paracoccidioidomycosis (chronic adult form) using the Terasaki lymphocytotoxicity test modified by Amos for HLA antigen analysis. AIMS: To discover indications of whether or not individual susceptibility to P. brasiliensis might depend on some specific immunological defect. RESULTS: There is no evidence of association between a specific HLA antigen and paracoccidioidomycosis in the subjects studied. Further investigations are recommended. PMID- 11032059 TI - Bath psoralen-ultraviolet A therapy in atopic eczema. AB - AIMS: To evaluate the efficacy of bath psoralen-ultraviolet A (PUVA) therapy in severe cases of atopic dermatitis (AD) in adults using the extended Six Area, Six Sign Atopic Dermatitis (SASSAD) score. METHODS: Thirty-five adult subjects with severe AD underwent bath PUVA therapy for a maximum of 30 sessions ranging from one to three times a week. The only other treatment allowed during the study was topical application of hydrocortisone. RESULTS: There were a total of six drop outs, three due to aggravation of symptoms. After the maximum 30 sessions the remaining 29 subjects showed 82.1% improvement in the severity of lesions, 75.2% reduction in extension of lesions, 74% improvement in itching and 79% improvement in night-time rest. Patient evaluations gave an overall score of 8.8 on a scale of 0-10. PMID- 11032060 TI - Chronic urticaria: diagnostic recommendations. AB - A group of European recognized dermatologists - the European Working Group on the Diagnosis of Chronic Urticaria - met on 12 March 1999 to discuss best practice for assessing and diagnosing patients with chronic urticaria. These are their recommendations. The recommendations will be valuable for the majority of cases, but are not exhaustive and may not include every possible precipitating factor. PMID- 11032061 TI - Aetiological factors in vulvar dermatitis. AB - AIM: To determine aetiological factors in vulvar dermatitis. BACKGROUND: Dermatitis of the vulva is a common clinical diagnosis in the specialized vulva clinic, although the exact prevalence is unknown. Aetiological factors are poorly understood in particular the incidence of atopy and seborrhoeic dermatitis. Other factors implicated are iron deficiency and exogenous causes such as contact irritants and allergens. METHODS: During a 1-year period, 38 patients with an initial clinical diagnosis of vulvar dermatitis were questioned, examined and investigated with full blood count, serum ferritin, bacterial swab where indicated and punch biopsy. After three patients had been excluded with other diagnoses, prick testing and patch testing were carried out. RESULTS: Of the 35 patients with vulvar dermatitis, 34 (97%) had either an atopic background and/or seborrhoeic dermatitis. Iron deficiency occurred in seven patients (20%) and relevant positive patch tests in nine patients (26%). Twenty patients had biopsies and all had confirmatory histology. CONCLUSION: This study suggests that the main aetiological factor in vulvar dermatitis is an endogenous predisposition. Iron deficiency may also contribute, while contact with irritants and allergens may explain the localization and frequently complicates and exacerbates the picture. Secondary sensitization to multiple substances is common. PMID- 11032062 TI - Treatment of recalcitrant ulcers in pyoderma gangrenosum with mycophenolate mofetil and autologous keratinocyte transplantation on a hyaluronic acid matrix. AB - Pyoderma gangrenosum sometimes takes a recalcitrant course that is unresponsive to standard immuno-suppression with corticosteroids and/or cyclosporin A. In these cases improvement of painful ulcerations is a therapeutic challenge. We report a 17-year-old boy with severe pyoderma gangrenosum treated successfully with mycophenolate mofetil and autologous keratinocyte transplantation using an esterified hyluronic acid delivery system. PMID- 11032063 TI - Generalized morphoea in a patient with Felty's syndrome. AB - We describe a 54-year-old woman with long-standing rheumatoid arthritis complicated by Felty's syndrome and lung involvement who developed generalized morphoea with no features of systemic sclerosis. To our knowledge this is the first description of such a case. PMID- 11032064 TI - Amiodarone pigmentation, eye and thyroid alterations. AB - Amiodarone is an effective cardiac antiarytmic drug. Long-term, high dose use of the drug is associated with skin discolouration, corneal deposition and alterations in thyroid hormone levels. We present the case of a 61-year-old woman suffering from the combination of these three side-effects. PMID- 11032065 TI - Granulomatous mycosis fungoides. Report of two cases and review of the literature. AB - BACKGROUND: Granulomatous mycosis fungoides is an extremely rare type of cutaneous T-cell lymphoma. Two cases are described and checked for clinical, histological and therapeutic differences to other variants of mycosis fungoides. CASE REPORTS: Case 1: a 52-year-old patient with livid plaques covering the whole integument. Histological findings showed granulomas with multinuclear giant cells in addition to a malignant lymphohistiocytic infiltrate (monodonal T-cell receptor (TCR)-gamma rearrangement). Despite various chemotherapeutic regimens, progression to tumour stage was observed. Case 2: an 88-year-old man with plaques and ulcerating tumours on the trunk and head. Histological findings showed malignant T-lymphocyte infiltrate (monoclonal TCR-gamma rearrangement) and granulomas with multinuclear foreign-body giant cells. Complete regression of all lesions was achieved using both local psoralen-ultraviolet A and electron radiotherapy. CONCLUSIONS: The diagnosis of a granulomatous mycosis fungoides depends exclusively on the histological demonstration of granulomas. Distinct clinical characteristics are not present. Apart from granuloma formation, no other noticeable histological features are evident. The presence of granulomas in mycosis fungoides does not have prognostic implications, as cases with aggressive, but also with a prolonged course have been described. PMID- 11032066 TI - Erythema multiforme along Blaschko's lines. AB - A case of erythema multiforme along Blaschko's lines is reported in a 20-year-old female suffering from recurrent herpes labialis. Histological examination was compatible with the clinical features. Spontaneous resolution followed in 4 weeks without therapy. PMID- 11032068 TI - Darier's disease complicated by Kaposi's varicelliform eruption due to herpes simplex virus. AB - Darier's disease is an autosomal dominant dermatosis. It may rarely be complicated by localized or widespread cutaneous viral infections. We describe a case of Kaposi's varicelliform eruption due to herpes simplex virus type 1 presenting in a patient with Darier's disease. PMID- 11032067 TI - Florid cutaneous and mucosal papillomatosis with acanthosis nigricans revealing a primary lung cancer. AB - This is the report of an 80-year-old patient with diffuse brownish hyperpigmentation and velvety thickening of the skin with onset 1 year before. Warty lesions on his limbs were present as well as papillomatous and verrucous lesions on his lips, mouth and eyelid conjunctivae with hyperkeratosis of the nipples. Biopsies, performed at different sites, showed histological pictures consistent with a diagnosis of acanthosis nigricans (AN) with florid cutaneous and mucosal papillomatosis. This type of AN is frequently associated with internal malignancy. In our patient serum levels of tissue polypeptide antigen, carcinoembryonic antigen, cytokeratin fragment and squamous cell carcinoma antigen were high and chest computed tomography scan indicated a large tumour infiltrating the right lung and extending to the mediastinum. Cytological examination of bronchial drainage revealed the presence of neoplastic cells, non small cell type carcinoma. The most frequent cancer associated with malignant AN is gastric adenocarcinoma. Lung tumour has rarely been reported with AN. Malignant AN is sometimes associated with other cutaneous and mucosal warty lesions, as in our patient. These various skin and mucosal lesions are the expression of a systemic epithelial disorder and may help clinicians to suspect a malignant form of AN. PMID- 11032069 TI - Nail changes in Langerhans cell histiocytosis. AB - Nail changes in Langerhans cell histiocytosis are distinctly uncommon. Paronychial erythema, swelling and subungual pustules of the fingernails and toenails were cardinal, and were supported by diffuse as well as dense collections of mononuclear Langerhans cells evidenced by microscopic investigation. Oral administration of co-trimoxazole (800 mg sulphamethoxazole + 160 mg trimethoprim) every 12 h, 50 mg/d cyclophosphamide and 80 mg/d predinisolone were the mainstay of treatment, supported by scalp tar shampoo and local betamethasone lotion application. PMID- 11032070 TI - Isolated granulocytic sarcoma of the skin in an elderly patient: good response to treatment with local radiotherapy and low-dose methotrexate. AB - Granulocytic sarcoma of the skin is frequently associated with haematological diseases and is rarely isolated. The disease generally develops into acute systemic myeloid leukaemia and is associated with a poor prognosis. We report an elderly patient presenting isolated granulocytic sarcoma of the skin who showed a very good response to treatment with local radiotherapy and low-dose methotrexate. PMID- 11032071 TI - Photosensitivity associated with treatment with triflusal. AB - BACKGROUND: Triflusal is a fluorinated aspirin derivative with antiplatelet properties, which is used in Spain for the management and prevention of thromboembolic disease. CASE REPORT: A 91-year-old female developed a systemic photosensitivity reaction 15 days after beginning triflusal preventive treatment (300 mg/12 h) for prior transient ischaemic attack. Photobiological study showed an abnormal response to light in areas exposed to ultraviolet B and A radiation, with a photopatch test positive to both triflusal and its main metabolite. These observations suggested a causal relation between triflusal and the clinical findings, as described in previous reports. CONCLUSIONS: The few cases reported to date and the clinicopathological features of this case suggested an immunological response as the most likely cause of the reaction. PMID- 11032073 TI - Comparative chemical evaluation of derivatives of hyaluronic acid. PMID- 11032072 TI - Fish skin-induced dermatitis. PMID- 11032074 TI - Endoscopic examination in Kaposi's sarcoma. PMID- 11032075 TI - Metastatic breast carcinoma: diffuse involvement of the skin. PMID- 11032076 TI - Dermatomyositis-like eruption following hydroxyurea therapy. PMID- 11032077 TI - Generalized Trichophyton rubrum infection in a subject with pemphigus vulgaris. PMID- 11032078 TI - Squamous cell carcinoma in chronic trophic ulcers of leprosy patients. PMID- 11032079 TI - Echographic test and cyclosporin therapy in psoriatic arthritis. PMID- 11032080 TI - Acute appendicitis and isotretinoin. A coincidence? PMID- 11032081 TI - Internal organ lesions in acrosclerosis patients: interleukin-2 and soluble receptor for interleukin-2 plasma levels. PMID- 11032082 TI - Where are we now and where do we go next in terms of the scientific basis for regulation on bioavailability and bioequivalence? FDA Biopharmaceutics Coordinating Committee. PMID- 11032083 TI - The basis for individual bioequivalence. FDA Population and Individual Bioequivalence Working Group. PMID- 11032084 TI - Dissolution and in vivo evidence of differences in reference products: impact on development of generic drugs. AB - The WHO List of International Comparator Pharmaceutical Products (CPP) For Equivalence Assessment of Interchangeable Multi-Source (Generic) Products will address an important issue in developing new generic drugs because it will identify the 'correct' reference product. This list will reduce unnecessary clinical studies in jurisdictions requiring new generics to be compared with brand products sold locally. Eventually, by employing the CPP, there will be a world-wide standard for brand and generic drugs, assuring the same level of quality internationally. The strategy of a single global reference is meritorious, but there are several hurdles to overcome. Most important is that the same brand may differ in dissolution and/or bioavailability in various jurisdictions, including some drugs with a narrow therapeutic index like phenytoin. Several examples are provided in this manuscript. This issue of regional differences has relevance, not only to the WHO list, but also to the matter of how safety and efficacy was established for that product in the first place. Normally, phase III clinical studies are conducted on a product manufactured in a single site, set to one standard. If the product differs in bioavailability in different jurisdictions, one is left with the question: 'which product has remained true to the original formulation?' Alternatively, if safety and efficacy is maintained with all formulations, then one is faced with the question: 'are the criteria currently employed for bioequivalence unnecessarily restrictive?' PMID- 11032085 TI - Guidances related to bioavailability and bioequivalence: European industry perspective. AB - The investigations of bioavailability and bioequivalence can be classified according to three separate areas of information. Firstly, estimation of bioavailability judged on a drug substance's in vivo characteristics taking into account solubility, polymorphism, stability (especially under the conditions of the GI tract), gut wall permeability and first pass metabolism. Secondly, evaluation of formulation properties including dissolution profile in the GI tract and its contribution to exposure variability with respect to the desired absorption characteristics. Finally, maintaining quality during the market phase with respect to equivalence to the clinical trial formulations. While in the first two areas, the range of the estimated mean values and the intra- and inter subject variabilities contain the desired information for proper medical decisions, in the third area the mean values and their confidence limits describe the quality with regard to the formulations of proven efficacy. Guidelines should clearly distinguish between the different areas in their recommendations regarding the intended information, e.g. mean values and/or ranges and confidence intervals. New approaches of granting limited waivers for BE studies (e.g. Biopharmaceutical Classification System (BCS)) should be expanded to consideration of pharmacokinetic properties of drugs (e.g. gastrointestinal metabolism, evidence for an absorption window, magnitude of first-pass effect, half-life) as already partly implemented in the German waiver concept, and further (scientifically) validated to achieve world-wide harmonisation (e.g. via ICH). PMID- 11032086 TI - Japanese guidance on bioavailability and bioequivalence. AB - The guideline of bioequivalence tests for generic drugs in our country was recently revised (1), and guidelines for formulation change and different strengths were published in February 2000. This paper describes those guidelines briefly. PMID- 11032087 TI - Differences in reference products: dissolution and in vivo evidence. PMID- 11032088 TI - Current status of the international comparator (reference) product system. AB - Since 1993, WHO's Division of Drug Management and Policies has been working with drug regulatory authorities from some 20 countries, the pharmaceutical industry- including both research-based as well as generic industry--and academia to set up requirements and specifications for multisource (generic) pharmaceutical products (1). One important area has been the development of an international list of comparator (reference) products for equivalence testing. This has become increasingly necessary as the manufacture, regulatory approvals and use of the generic pharmaceutical products have increased world-wide. In order to save time and resources in product development and to avoid duplication of unnecessary bioavailability testing in humans, it has been suggested that an internationally agreed comparator product system would be helpful. Moreover, any exposure of a human being to a pharmaceutical product in equivalence testing may carry a health risk and may consequently be considered unethical. PMID- 11032089 TI - Equivalence concepts in clinical trials. AB - According to the recent ICH E9 Guidance Statistical Principles for Clinical Trials, efficacy is most convincingly established by demonstrating superiority to placebo, by showing superiority to an active control treatment or by demonstrating a dose-response relationship (so-called 'superiority' trials). For serious illnesses, a placebo-controlled trial may be considered unethical if a therapeutic treatment exists which has proven efficacious in relevant superiority trial(s). In that case, the scientifically sound use of an active treatment as a control should be considered. Active control trials designed to show that the efficacy of an investigational product is not relevantly worse than that of the active comparator are called 'non-inferiority' trials (1). After having confirmed non-inferiority, superiority of the alternative test treatment over the reference treatment can additionally be tested without the need to adjust the significance level (2). In contrast to cross-over bioequivalence trials based on pharmacokinetic endpoints such as AUC and Cmax, therapeutic equivalence and non inferiority trials are based on clinical end-points. Therefore, they are often conducted as parallel group comparisons. It is important to note that the conclusion of equivalence or non-inferiority is based on the inclusion of the appropriate confidence interval in the equivalence acceptance range, and that it cannot be derived from a non-significant test result of the inappropriate null hypothesis of no treatment difference. PMID- 11032090 TI - Biopharmaceutical characterisation of herbal medicinal products: are in vivo studies necessary? AB - Herbal medicinal products have to meet comparable standards concerning the assessment of efficacy, safety and (bio)pharmaceutical quality as chemically defined synthetic drugs. However, these requirements are not fulfilled for many herbal products so far, particularly regarding in vitro dissolution and in vivo bioavailability. The necessity of in vivo studies for a biopharmaceutical characterisation of the products depends on the solubility/permeability properties of the active drug ingredient as well as dissolution behaviour of the dosage form. Also, in the case of herbal medicinal products, a waiver of in vivo BA/BE studies is recommended as long as the active ingredient is highly soluble according to the Biopharmaceutics Classification System and dissolution of the dosage form takes place rapidly (>85%/20 min) in physiological buffer systems (pH 1-8). PMID- 11032091 TI - Pharmacometrics: modelling and simulation tools to improve decision making in clinical drug development. AB - There is broad recognition within the pharmaceutical industry that the drug development process, especially the clinical part of it, needs considerable improvement to cope with rapid changes in research and health care environments. Modelling and simulation are mathematically founded techniques that have been used extensively and for a long time in other areas than the pharmaceutical industry (e.g. automobile, aerospace) to design and develop products more efficiently. Both modelling and simulation rely on the use of (mathematical and statistical) models which are essentially simplified descriptions of complex systems under investigation. It has been proposed to integrate pharmacokinetic (PK) and pharmacodynamic (PD) principles into drug development to make it more rational and efficient. There is evidence from a survey on 18 development projects that a PK/PD guided approach can contribute to streamline the drug development process. This approach extensively relies on PK/PD models describing the relationships among dose, concentration (and more generally exposure), and responses such as surrogate markers, efficacy measures, adverse events. Well documented empirical and physiologically based PK/PD models are becoming available more and more, and there are ongoing efforts to integrate models for disease progression and patient behavior (e.g. compliance) as well. Other types of models which are becoming increasingly important are population PK/PD models which, in addition to the characterization of PK and PD, involve relationships between covariates (i.e. patient characteristics such as age, body weight) and PK/PD parameters. Population models allow to assess and to quantify potential sources of variability in exposure and response in the target population, even under sparse sampling conditions. As will be shown for an anticancer agent, implications of significant covariate effects can be evaluated by computer simulations using the population PK/PD model. Stochastic simulation is widely used as a tool for evaluation of statistical methodology including for example the evaluation of performance of measures for bioequivalence assessment. Recently, it was suggested to expand the use of simulations in support of clinical drug development for predicting outcomes of planned trials. The methodological basis for this approach is provided by (population) PK/PD models together with random sampling techniques. Models for disease progression and behavioral features like compliance, drop-out rates, adverse event dependent dose reductions, etc. have to be added to population PK/PD models in order to mimic the real situation. It will be shown that computer simulation helps to evaluate consequences of design features on safety and efficacy assessment of the drug, enabling identification of statistically valid and practically realisable study designs. For both modelling and simulation a guidance on 'best practices' is currently worked out by a panel of experts comprising representatives from academia, regulatory bodies and industry, thereby providing a necessary condition that model-based analysis and simulation will further contribute to streamlining pharmaceutical drug development processes. PMID- 11032092 TI - Cluster of HIV-infected adolescents and young adults--Mississippi, 1999. AB - From February through June 1999, seven human immunodeficiency virus (HIV) infected young persons were identified in a small town in rural Mississippi. Two persons were identified through routine voluntary HIV testing during sexually transmitted disease (STD) evaluations, and five were identified subsequently through contact investigation by the local health department. Contact investigation identified sex partners and social contacts (i.e., persons who shared social surroundings) and defined a social network of 122 sex and social contacts. Seven (9%) of 78 persons tested from the social network were HIV infected. Within the social network, a sexual contact network of 44 persons (the seven HIV-infected persons and their sex partners) was identified. The Mississippi State Department of Health asked CDC to join the investigation to describe further the cluster and help direct prevention efforts. This report summarizes the investigation of this cluster and underscores the need for HIV prevention and treatment in rural areas. PMID- 11032093 TI - Nutritional assessment of adolescent refugees--Nepal, 1999. AB - During 1990-1993, 83,000 ethnic Nepalese fled from Bhutan to refugee camps in southeast Nepal after new citizenship policies were enacted by the Bhutanese government. Although annual nutrition surveys of children aged <5 years had been conducted by international agencies, no anthropometric assessment of adolescents had been performed since the refugees arrived in 1990. After withdrawal of a fortified cereal from their rations, the number of reported cases of angular stomatitis (AS) (i.e., thinning and/or fissuring at the angles of the mouth, a sign of possible vitamin deficiency) increased six-fold during December 1998 March 1999 (from 5.5 to 35.6 cases per 1000 refugees) (Santa Tamang, MD, Save the Children Fund, United Kingdom, personal communication, 1999). The highest rates of AS were found among children and adolescents. In October 1999, CDC was invited by the World Food Programme and the United Nations High Commissioner for Refugees to assess the health status of adolescent refugees. This report summarizes the investigation, which indicated a high prevalence of low body mass index (BMI), anemia, low vitamin A status, and signs of micronutrient deficiencies among adolescent refugees. PMID- 11032094 TI - Progress toward poliomyelitis eradication--Ethiopia, 1997-August 2000. AB - In 1988, the World Health Assembly resolved to eradicate poliomyelitis globally by 2000 (1). Following the signing of the Yaounde Declaration on Polio Eradication in Africa in 1996, Ethiopia joined global efforts toward polio eradication (2). Since then, Ethiopia has accelerated implementation of polio eradication strategies. This report summarizes progress toward polio eradication in Ethiopia during 1997-August 2000 and highlights the remaining challenges toward achieving the goal. PMID- 11032095 TI - The increasing impact of arthritis on public health. AB - Age is the most powerful risk factor for osteoarthritis (OA) in the United States. It is estimated that 68% of individuals older than 55 years have radiographic evidence of OA. The US is growing older--the over-65 age group represented only 4% of the population in 1900, but accounted for 12.4% in 1988, and is projected to account for 22% by the year 2030. As the age of our population has increased, so has the prevalence of arthritis. About 43 million individuals (1 in 6) have arthritis, and most are older than 45 years. By the year 2020, 59.4 million persons in the US will be affected by arthritis, thus increasing chronic disability and costs by more than 25%. The annual cost to society in medical care and lost wages is currently estimated to be $65 billion, and is projected to escalate to $95 billion by the year 2000. Physicians who provide care for the increased number of patients with arthritis in the 21st century must be aware of improved therapeutic modalities to reduce arthritis related disabilities, hospitalizations, and complications related to therapy, to minimize the risk of adverse drug reactions, and to preserve function. PMID- 11032096 TI - From prostaglandin replacement to specific COX-2 inhibition: a critical appraisal. AB - Nonsteroidal antiinflammatory drugs (NSAID) are among the most widely prescribed medications worldwide. While these agents are efficacious for the relief of both pain and the signs and symptoms of inflammation, the use of traditional NSAID is associated with a high prevalence of toxic events. It is estimated that 2-4% of traditional NSAID users will have a serious adverse gastrointestinal (GI) event per year, with a point prevalence of GI ulcers at any given time of 10-30%. The mortality rate associated with these NSAID is one of the highest attributed to the use of any agent. Prostaglandin replacement with misoprostol has been shown to significantly reduce the incidence of serious GI toxicity related to the use of conventional NSAID. All conventional NSAID inhibit both cyclooxygenase-1 (COX 1) and COX-2 enzymes. As COX-1 is constitutively expressed in many tissues and contributes to the maintenance of the normal physiologic state in the GI tract, the kidney, and platelets, an agent that specifically inhibits COX-2, an inducible enzyme associated with the inflammatory response, would be highly desirable. Specific COX-2 inhibition would have the advantage of improving the signs and symptoms of inflammation, while avoiding the possibly significant toxicity of traditional NSAID. This article describes the biology of eicosanoids and the effects of NSAID, the toxicity profile of conventional NSAID, and the rationale for the use of gastroprotective agents or agents that specifically inhibit the COX-2 isozyme. PMID- 11032097 TI - Defining COX-2 inhibitors. AB - Arachidonic acid metabolism is governed by 2 isoforms of cyclooxygenase (COX), the constitutively expressed COX-1 and the inducible COX-2. Antiinflammatory, analgesic, and antipyretic effects of nonsteroidal antiinflammatory drugs (NSAID) are explained by the capacity of these agents to inhibit COX-2, whereas the serious gastrointestinal side effects are caused by inhibitors of COX-1. The first of a new class of COX inhibitors, the COX-2 specific inhibitors, has just been approved for the treatment of osteoarthritis (OA) and rheumatoid arthritis (RA). As the clinical outcomes of specific COX-2 inhibitors are considerably different than those of NSAID, it is essential for the clinician to understand the basis of classification of those new, effective, and safer therapeutic agents for the treatment of OA and RA. PMID- 11032098 TI - Antiinflammatory and analgesic efficacy of COX-2 specific inhibition: from investigational trials to clinical experience. AB - Research strongly indicates that increased expression of the isoenzyme cyclooxygenase-2 (COX-2) is responsible for elevated production of prostaglandins in inflamed joint tissues and is involved in the mediation of pain. In contrast, COX-1 is a constitutively produced isoenzyme that is involved in the synthesis of eicosanoids that have important homeostatic functions, for example, in the gastric mucosa and platelets. This new knowledge led to the development of drugs that are highly specific inhibitors of COX-2 while not inhibiting COX-1 at maximally efficacious dosages. The first COX-2 specific agent approved for clinical use in the United States was celecoxib. Large multicenter trials have shown that celecoxib at dosages of 100 mg BID and 200 mg BID is as effective as naproxen 500 mg BID in patients with osteoarthritis of the knee or hip. Another large multicenter trial also demonstrated that celecoxib 200 mg BTD and 400 mg BID is as effective as naproxen 500 mg BID in patients with rheumatoid arthritis (RA). A comparative trial showed that celecoxib 200 mg BID is as effective as diclofenac SR 75 mg BID in patients with RA. The potential of COX-2 specific inhibitors to provide antiinflammatory and analgesic efficacy equivalent to that of conventional nonsteroidal antiinflammatory drugs without the adverse gastrointestinal mucosal and platelet effects associated with nonspecific COX inhibitors promises to revolutionize the clinical care of arthritis patients. PMID- 11032099 TI - Significant upper gastrointestinal events associated with conventional NSAID versus celecoxib. AB - Despite their substantial clinical benefits in the management of rheumatoid arthritis, osteoarthritis, pain, and other musculoskeletal complaints, conventional nonsteroidal antiinflammatory drugs (NSAID) are associated with significant toxicities that can frequently limit their use. The most common and noteworthy adverse effects of NSAID are gastrointestinal (GI), and range from dyspeptic symptoms to ulcers and serious ulcer complications. The upper GI toxicities associated with the use of conventional NSAID led to the search for medications that were as clinically effective as these agents, but with a significantly improved GI safety profile. It is now known that the constitutively expressed isoenzyme cyclooxygenase (COX)-1 catalyzes the synthesis of prostanoids that help to regulate normal physiologic processes, including GI mucosa protection, whereas the inducible isoenzyme COX-2 leads to the generation of prostaglandins that mediate inflammation, pain, and fever. This knowledge has led to the development of new compounds that, at therapeutic concentrations, inhibit COX-2 without affecting COX-1. The first COX-2 targeted agent approved by the US Food and Drug Administration (FDA) was celecoxib. This article reviews the risks of GI complications associated with conventional NSAID use and compares these risks with that of the new COX-2 specific inhibitor celecoxib. PMID- 11032100 TI - Football, or soccer, as it is called in North America, is the most popular sport worldwide. PMID- 11032101 TI - Football injuries and physical symptoms. A review of the literature. AB - Football is one of the most popular sports worldwide. The frequency of football injuries is estimated to be approximately 10 to 35 per 1000 playing hours. The majority of injuries occur in the lower extremities, mainly in the knees and ankles; the number of head injuries is probably underestimated. The average cost for medical treatment per football injury is estimated to be $150 (U.S. dollars). Considering the number of active football players worldwide, the socioeconomic and financial consequences of injury are of such a proportion that a prevention program to reduce the incidence of injuries is urgently required. For this reason, an analysis of intrinsic (person-related) and extrinsic (environment related) risk factors was undertaken based on a review of the current literature. It was concluded that the epidemiologic information regarding the sports medicine aspects of football injuries is inconsistent and far from complete because of the employment of heterogeneous methods, various definitions of injury, and different characteristics of the assessed teams. The aim of this study was to analyze the literature on the incidence of injuries and symptoms in football players, as well as to identify risk factors for injury and to demonstrate possibilities for injury prevention. PMID- 11032102 TI - The influence of psychological factors on sports injuries. Review of the literature. AB - The influence of psychological factors on sports injuries has been demonstrated in numerous empirical studies. Almost all investigations have been based on stress theory or a personality-profile approach. Although the majority of studies have employed different methods, the results are in general agreement that "life events" can influence the risk of injury in athletes. In this context, social support appears to have a buffering effect. According to existing results, the influence of stress-coping strategies is somewhat questionable. From the numerous psychological attributes that have been investigated in relation to sports injuries, only competitive anxiety has been shown to be associated with injury occurrence. A personality profile typical of the "injury-prone" athlete does not exist. However, several studies have shown a certain readiness to take risks (lack of caution, adventurous spirit) on the part of injured athletes. In this review, the current knowledge regarding the relationship between psychological factors and sports injuries is presented and a stress theory model is developed. PMID- 11032103 TI - Medical history and physical findings in football players of different ages and skill levels. AB - The physical and physiologic demands of football on its participants become more pronounced as the level of competition increases. The aim of this study was to compare the medical history and physical findings in players from different levels of play as well as to analyze the relationship between pathologic findings in the joints of the lower extremities and the medical history. Five hundred eighty-eight football players from eight age and skill levels were investigated. The medical histories of the players were obtained by questionnaire. A physical examination conducted by specially trained physicians incorporated anthropometric and body fat measurements and examination of the spine and the hip, knee, foot, and great toe joints. On average, the players reported 6.6 (SD, 8.8) previous injuries. At the time of the examination, 136 players (24%) still felt the effects of a previous injury. Almost one-quarter of the players (134, 23%) had a pathologic finding in either the right or left knee, and even more players (162, 28%) had a pathologic anterior drawer sign either in the right or left ankle. A correlation was found between the location of the pathologic findings and the preferred leg for playing football. We recommend that further research should address the prevalence of pathologic findings and complaints in football players as well as the secondary structural changes that may occur as the result of playing football. PMID- 11032104 TI - Psychological and sport-specific characteristics of football players. AB - It is hypothesized that players of different levels of play might differ not only in their football skills but also in their way of playing football and with respect to psychological factors such as concentration, reaction time, or competitive anxiety. The psychological characteristics of a player might influence his way of playing football (in particular with respect to fair play) and also his risk of injury. A group of 588 football players were studied by questionnaire; additionally, reaction time tests were performed. Psychological characteristics were assessed by three established self-evaluation questionnaires: the Athletic Coping Skills Inventory, the State Competitive Anxiety Test, and the State-Trait-Anger-Expression-Inventory. Football-specific characteristics that were investigated included playing experience and positions played, style of play, number of training hours and games, as well as aspects of fair play. Reaction time was tested twice: without the influence of physical exercise and immediately after a 12-minute run. A significant reduction in reaction time was observed after physical exercise. In high-level players, the reaction time immediately after the 12-minute run was significantly shorter than it was in low-level players. The questionnaire answers given regarding fair play clearly indicated that fair play is not paid sufficient respect. The relationship between psychological characteristics and attitudes toward fair play was analyzed and discussed. PMID- 11032105 TI - Assessment and evaluation of football performance. AB - The most important variables for measuring performance in team sports such as football are physical condition and technical and tactical performance. However, because of the complexity of the game of football it is difficult to ascertain the relative importance of each of these variables. The aim of the present study was to develop a standardized test battery to evaluate physical performance in football players. The F-MARC test battery was designed to closely relate to the football player's normal activity and comprised a functional, structured training session of approximately 2.5 hours. It included a "quality rating" of the warm-up procedure, tests of flexibility, football skills, power, speed, and endurance. The players finished with a cool-down. A total of 588 football players underwent the F-MARC test battery. Mean values for performance on each test are presented for groups of differing age and skill levels. The test battery proved to be a feasible instrument to assess both physical performance and football skills. This study supports the proposal by Balsom (1994) that analysis of an individual player's physical profile, in relation to mean values for a similar age group and skill level, might be of assistance to the coach in objectively evaluating the effects of a specific training program. It may also be of use to the physician and physical therapist responsible for monitoring progress during rehabilitation after football injuries. PMID- 11032106 TI - Influence of definition and data collection on the incidence of injuries in football. AB - Studies on the incidence of football injuries vary in the definition of injury, study design, methods of data collection, and observation periods. The aim of this study was to review the different methodologies applied in the evaluation of football injuries as well as to analyze the influence of data collection methods on the incidence of football injuries. In this study, injury data obtained weekly by a physician during 1 year of follow-up in 264 football players were compared with the results of retrospective questionnaires completed by the players at the end of the observation period. In the retrospective questionnaire, the incidence of injuries as well as of complaints was significantly lower than that found in the weekly follow-up examinations. Approximately every third moderate injury and less than 10% of the mild injuries were remembered retrospectively. The shorter the period of symptoms and the longer ago the injury occurred, the more frequently it was forgotten. However, even severe injuries, such as fractures, were not reported in the retrospective investigation. Based on the review of the literature and the data presented, recommendations in relation to study design, the definition of injury, and the calculation of incidence are proposed. PMID- 11032107 TI - Incidence of football injuries in youth players. Comparison of players from two European regions. AB - Several authors have investigated the frequency of football injuries in youth players. However, the results of these studies are inconsistent because of the different age groups investigated and the different methods applied. The aim of the present study was to compare the incidence and characteristics of football injuries in youth players of two European regions. A total of 444 youth players from the Czech Republic and the Alsace region of France and Germany were followed weekly for 1 year. In 311 players (70%), complete weekly follow-ups over the 1 year period were available. The comparison of injury data revealed no substantial differences between players from the Alsace region and the Czech Republic in injury incidence per 1000 hours of exposure, degree of injury severity, or the circumstances in which the injuries occurred. However, players from the Czech Republic spent more time in training and playing football than did players from the Alsace region, and in the Czech Republic a higher proportion of injuries was caused by foul play. With only a few exceptions, the statistics were similar in the amount of football played as well as in the incidence of injury between different age and skill levels in both European regions. PMID- 11032108 TI - Incidence of football injuries and complaints in different age groups and skill level groups. AB - In this study, the incidence of football injuries and complaints as related to different age groups and skill levels was studied over the period of 1 year. All injuries and complaints as well as the amount of time players spent in training and games were recorded. All injured players were examined weekly by physicians, and all injuries were assessed according to the International Classification of Diseases (ICD-10), which describes them in terms of injury type and location, the treatment required, and the duration of subsequent performance limitations. A total of 264 players of different age groups and skill levels was observed for 1 year. Five hundred fifty-eight injuries were documented. Two hundred sixteen players had one or more injuries. Only 48 players (18%) had no injury. The average number of injuries per player per year was 2.1. Injuries were classified as mild (52%), moderate (33%), or severe (15%). Almost 50% of all injuries were contact injuries; half of all the contact injuries were associated with foul play. The majority of injuries were strains and sprains involving the ankle, knee, and lumbar spine. Nearly all players (91%) suffered from complaints related to football. Only 23 players reported no injuries and no complaints. Prevention programs, fair play, and continuing education in techniques and skills may reduce the incidence of injuries over time. PMID- 11032109 TI - Severe injuries in football players. Influencing factors. AB - The aims of this prospective study were to analyze factors related to the occurrence of severe football injuries in players of different ages (14 to 42 years) and different skill levels (local teams to first league teams). In the Czech Republic, 398 players were followed up for 1 year, during which time they sustained 686 injuries. Of these, 113 (16.5%) were severe injuries. Ninety-seven severe injuries (86%) were able to be documented in detail. Trauma was the cause of 81.5% of the injuries and overuse was the cause of 18.5%. Joint sprains predominated (30%), followed by fractures (16%), muscle strains (15%), ligament ruptures (12%), meniscal tears and contusions (8%), and other injuries. Injuries to the knee were most prevalent (29%), followed by injuries to the ankle (19%) and spine (9%). More injuries occurred during games (59%) than in practice. Twenty-four percent of the injured players had suffered a previous injury of the same body part. Forty-six percent of injuries were caused by contact and 54% involved no body contact. Thirty-one percent of severe injuries were caused by foul play. From these results and the analysis of injuries in specific body parts, the following factors were determined to influence the occurrence of severe injuries: 1) personal factors (intrinsic): age of player, previous injuries, joint instability, abnormality of the spine, poor physical condition, poor football skills, or inadequate treatment and rehabilitation of injuries; 2) environmental factors (extrinsic): subjective exercise overload during practices and games, amount and quality of training, playing field conditions, equipment (wearing of shin guards and taping) and violations of existing rules (foul play). PMID- 11032110 TI - Risk factor analysis for injuries in football players. Possibilities for a prevention program. AB - Review of the literature shows that information concerning risk factors for football injuries is incomplete and partly contradictory. The aim of this study was to analyze the influence of medical history, physical findings, football skills, and football performance, as well as psychosocial characteristics on the occurrence and severity of football injuries. The prospective outline of the study was as follows: after a baseline examination was performed to ascertain possible predictors of injury, all players were followed up weekly for 1 year to register subsequent injuries and complaints. Two hundred sixty-four of 398 players (67%) had complete weekly follow-ups over 1 year. A majority of the players (N = 216; 82%) were injured during the observation period. In comparing injured and uninjured players, several differences were observed. To create a multidimensional predictor score for football injuries, 17 risk factors were selected. These risk factors covered a wide spectrum, such as previous injuries, acute complaints, inadequate rehabilitation, poor health awareness, high life event stress, playing characteristics, poor reaction time, poor endurance, and insufficient preparation for games. By summing up the individual risk factors, a predictive sum was calculated for each player. The more risk factors present at the baseline examination, the higher the probability of that player incurring an injury in the ensuing year. Using two risk factors as the cut-off score, more than 80% of the players were correctly classified as to whether they went on to incur an injury. Based on these findings, knowledge from the literature, and practical experience, possibilities for a prevention program are suggested. PMID- 11032111 TI - Assessing water quality impacts and cleanup effectiveness in streams dominated by episodic mercury discharges. AB - Accurate pollutant mass budgets are needed for identifying contaminant sources and establishing cleanup goals. We monitored mercury discharges from an abandoned mine site in northern California with the objectives of: (1) estimating the mass loading of mercury from the site; (2) evaluating the factors that control the mercury discharges; (3) assessing the significance of peak flows in transporting contaminants; and (4) developing methods for measuring the effectiveness of cleanup efforts. We sampled water downstream from the mine site over a wide range of streamflows. Mercury concentrations varied over 2000-fold, from 485 to 1040000 ng/l, grossly exceeding the regulatory water quality objective of 12 ng/l at all times. Particulate mercury represented over 99.97% of the total mercury, and mercury concentrations were closely correlated to suspended sediment concentrations (r = 0.98). Thus, we can use suspended sediment concentrations as a proxy for mercury concentrations, and calculate a continuous record of mercury flux from continuous monitoring of streamflow (using a small flume) and turbidity (using an optical backscatter sensor). Mercury fluxes inferred in this way are consistent with fluxes estimated from field samples. In January and February of 1998, our small abandoned mine site released approximately 82 kg of mercury to downstream waters. Most of the mercury was released during brief intense rainstorms. For example, in one 200-min period we recorded 3.4 cm of rain, a 2.6 fold increase in streamflow (460-1120 l/s), and an 82-fold increase in mercury flux (1.2-99 g/min). Over 75% of the total mercury flux during this 2-month period occurred in less than 10% of the total time. In systems such as this one, where contaminant transport is highly episodic, sampling programs that miss the high-flow episodes may greatly underestimate the actual water quality threat. In addition, changes in pollutant fluxes or concentrations in receiving waters may not reflect changes in pollutant sources (such as an environmental cleanup) if the stochastic forcing (e.g. intense rainstorms) varies through time. We propose that water quality trends can be more accurately measured by changes in the relationship between contaminant flux and stochastic driving factors, as expressed by contaminant rating curves. PMID- 11032112 TI - Do concepts about catchment cycling of methylmercury and mercury in boreal catchments stand the test of time? Six years of atmospheric inputs and runoff export at Svartberget, northern Sweden. AB - Previous studies at the Svartberget catchment in northern Sweden have identified potential terrestrial sources of methylmercury (MeHg) and total mercury (THg) in runoff as well as processes controlling MeHg/Hg transfers from soil to runoff water. This paper considers whether the concepts based on a few seasons of observations are consistent with catchment budgets of MeHg/THg over half a decade. Inter-annual and seasonal variations in the input and output fluxes of THg/MeHg, in open field wet deposition (OF), litterfall (LF) and runoff water are evaluated together with more recently measured concentrations that include the throughfall water (TF) data. The input and output flux data of THg and MeHg from the Svartberget catchment are also compared to those from the Gardsjon Catchment. The average annual MeHg input fluxes in OF, TF and LF are 0.08, 0.17 and 0.3 g km(-2) year(-1), respectively. The comparable inputs for THg are 7, 15 and 17 g km(-2) year(-1). Thus, LF is as important as TF for THg inputs, while LF is twice as important as TF for MeHg inputs. The annual output flux of MeHg varied between 0.05 and 0.14 g km(-2) year(-1). The annual output flux of THg varied between 1 and 3.4 g km(-2) year(-1). The large inter-annual variations in catchment output did not follow the smaller variations in atmospheric input. This suggests that changes in climate can effect terrestrial outputs of THg/MeHg to surface water more than atmospheric deposition. These data do not contradict the earlier findings that it is the hydrological and biogeochemical processes in the riparian zone that have a central role in determining the amount of MeHg reaching surface waters from forested catchments. The juxtaposition of major flow paths and organic-rich soils in the riparian zone may create the locations of most importance for net MeHg production, runoff export and a larger pool of MeHg. PMID- 11032113 TI - Distribution, speciation, and transport of mercury in stream-sediment, stream water, and fish collected near abandoned mercury mines in southwestern Alaska, USA. AB - Concentrations of total Hg, Hg (II), and methylmercury were measured in stream sediment, stream-water, and fish collected downstream from abandoned mercury mines in southwestern Alaska to evaluate environmental effects to surrounding ecosystems. These mines are found in a broad belt covering several tens of thousands of square kilometers, primarily in the Kuskokwim River basin. Mercury ore is dominantly cinnabar (HgS), but elemental mercury (Hg degrees) is present in ore at one mine and near retorts and in streams at several mine sites. Approximately 1400 t of mercury have been produced from the region, which is approximately 99% of all mercury produced from Alaska. These mines are not presently operating because of low prices and low demand for mercury. Stream sediment samples collected downstream from the mines contain as much as 5500 microg/g Hg. Such high Hg concentrations are related to the abundance of cinnabar, which is highly resistant to physical and chemical weathering, and is visible in streams below mine sites. Although total Hg concentrations in the stream-sediment samples collected near mines are high, Hg speciation data indicate that concentrations of Hg (II) are generally less than 5%, and methylmercury concentrations are less than 1% of the total Hg. Stream waters below the mines are neutral to slightly alkaline (pH 6.8-8.4), which is a result of the insolubility of cinnabar and the lack of acid-generating minerals such as pyrite in the deposits. Unfiltered stream-water samples collected below the mines generally contain 500-2500 ng/l Hg; whereas, corresponding stream-water samples filtered through a 0.45-microm membrane contain less than 50 ng/l Hg. These stream-water results indicate that most of the Hg transported downstream from the mines is as finely-suspended material rather than dissolved Hg. Mercury speciation data show that concentrations of Hg (II) and methylmercury in stream water samples are typically less than 22 ng/l, and generally less than 5% of the total Hg. Muscle samples of fish collected downstream from mines contain as much as 620 ng/g Hg (wet wt.), of which 90-100% is methylmercury. Although these Hg concentrations are several times higher than that in fish collected from regional baseline sites, the concentration of Hg in fish is below the 1000 ng/g action level for edible fish established by the US Food and Drug Administration (FDA). Salmon contain less than 100 ng/g Hg, which are among the lowest Hg contents observed for fish in the study, and well below the FDA action level. PMID- 11032114 TI - Mercury in the River Nura and its floodplain, Central Kazakhstan: I. River sediments and water. AB - The River Nura in Central Kazakhstan has been heavily polluted by mercury originating from an acetaldehyde plant. Mercury in the riverbed is mainly associated with power station fly ash, forming a new type of technogenic deposit. A systematic survey of the bed was carried out to establish the location, extent and nature of the contaminated sediments, and to evaluate the potential for sediment transport. The bed sediments were found to contain very high concentrations of mercury, particularly in the first 15 km downstream of the source of the pollution. Average total mercury concentrations in this section of the river are typically between 150 and 240 mg/kg, falling rapidly with increasing distance downstream. The estimated total volume of silts in the riverbed between Temirtau, the origin of the pollution, and Intumak Reservoir, located 75 km downstream, has been calculated as 463500 m3, containing an estimated 9.4 tonnes mercury. Forty-six percent of the total volume of contaminated silts containing almost 95% of the mercury are located in the upper 25 km of the river, however. The data clearly support the hypothesis that large quantities of polluted sediment are not transported long distances downstream but are removed from the aquatic environment in times of flood and deposited on the low-lying lands adjacent to the river. This process, however, does not stop mercury moving further downstream in the water column. PMID- 11032115 TI - Mercury in the River Nura and its floodplain, Central Kazakhstan: II. Floodplain soils and riverbank silt deposits. AB - A unique and serious case of mercury pollution has occurred in the River Nura and its floodplain in Central Kazakhstan, where mercury-rich wastewater from an acetaldehyde plant was discharged largely without treatment for several decades. In the river, the mercury became associated with millions of tonnes of power station fly ash, forming a new type of deposit known as 'technogenic silt'. During spring floods these highly contaminated silts are transported downstream and are dispersed over the floodplain, leading to widespread contamination of the land. A detailed survey of the floodplain was carried out to investigate the extent of pollution and to assess the need for remediation. Total mercury concentrations in the topsoils of the floodplain ranged from near background levels to over 100 mg/kg. Mercury concentrations in river bank deposits were found to range from a mean of 73.3 mg/kg Hg in the most contaminated section of the river to a mean of 13.4 mg/kg Hg at a distance of 70 km downstream. Concentrations were lower than corresponding concentrations in the riverbed within the first 25 km from the source of the pollution, but thereafter they were significantly higher. The results show that over the past 30-40 years a large proportion of the contaminated sediments from the river was deposited on the 70 km of banks and in the floodplain below the pollution source. Topsoils of the floodplain and silt deposits located on or close to the river banks contain an estimated 53 t and 65 t of mercury respectively, with an additional 62 t in a small natural swamp which was formerly used as a waste disposal area. The contamination is serious but relatively localized, with > 70% of the total amount of mercury in topsoils and > 90% of mercury in river bank deposits located within 25 km from the source. PMID- 11032116 TI - Mercury mine drainage and processes that control its environmental impact. AB - Mine drainage from mercury mines in the California Coast Range mercury mineral belt is an environmental concern because of its acidity and high sulfate, mercury, and methylmercury concentrations. Two types of mercury deposits are present in the mineral belt, silica-carbonate and hot-spring type. Mine drainage is associated with both deposit types but more commonly with the silica-carbonate type because of the extensive underground workings present at these mines. Mercury ores consisting primarily of cinnabar were processed in rotary furnaces and retorts and elemental mercury recovered from condensing systems. During the roasting process mercury phases more soluble than cinnabar are formed and concentrated in the mine tailings, commonly termed calcines. Differences in mineralogy and trace metal geochemistry between the two deposit types are reflected in mine drainage composition. Silica-carbonate type deposits have higher iron sulfide content than hot-spring type deposits and mine drainage from these deposits may have extreme acidity and very high concentrations of iron and sulfate. Mercury and methylmercury concentrations in mine drainage are relatively low at the point of discharge from mine workings. The concentration of both mercury species increases significantly in mine drainage that flows through and reacts with calcines. The soluble mercury phases in the calcines are dissolved and sulfate is added such that methylation of mercury by sulfate reducing bacteria is enhanced in calcines that are saturated with mine drainage. Where mercury mine drainage enters and first mixes with stream water, the addition of high concentrations of mercury and sulfate generates a favorable environment for methylation of mercury. Mixing of oxygenated stream water with mine drainage causes oxidation of dissolved iron(II) and precipitation of iron oxyhydroxide that accumulates in the streambed. Both mercury and methylmercury are strongly adsorbed onto iron oxyhydroxide over the pH range of 3.2-7.1 in streams impacted by mine drainage. The dissolved fraction of both mercury species is depleted and concentrated in iron oxyhydroxide such that the amount of iron oxyhydroxide in the water column reflects the concentration of mercury species. In streams impacted by mine drainage, mercury and methylmercury are transported and adsorbed onto particulate phases. During periods of low stream flow, fine-grained iron hydroxide sediment accumulates in the bed load of the stream and adsorbs mercury and methylmercury such that both forms of mercury become highly enriched in the iron oxyhydroxide sediment. During high-flow events, mercury- and methylmercury enriched iron hydroxide sediment is transported into larger aquatic systems producing a high flux of bioavailable mercury. PMID- 11032117 TI - Mercury distribution in waters and fishes of the upper Madeira rivers and mercury exposure in riparian Amazonian populations. AB - In this paper, the results of mercury concentrations in two abiotic compartments (river water and suspended particles) and two biotic compartments (fish and human hair) from the upper Madeira rivers of the Bolivian Amazon basin are presented. Because of the local hydrological regimes and a high deposition rate in the plain, due to the presence of a subsidence zone at the bottom of the Andean piedmont, in the dry season, the highest mercury concentrations and fluxes were not found in rivers where mining activities took place (2.25-6.99 ng l(-1); and 1.07-8.67 mg Hg d(-1) km(-2)), but at the outlet of the Andean basins exploited for their alluvial gold (7.22-8.22 ng l(-1); and 9.47-9.52 mg Hg d(-1) km(-2)). The total mercury concentrations measured in surface waters of the upper Beni basin varied during the dry season, from 2.24 to 2.57 ng l(-1) in the glacial waters of the Zongo river, to 7.00 ng l(-1) in the Madeira River at Porto Velho and 9.49-10.86 ng l(-1) at its confluence with the Amazon. The results obtained from fish indicate, on one hand, that 86% of the piscivorous fishes collected in the Beni river were contaminated, and, on the other hand, their high mercury concentrations could exceed by almost four times the WHO (1976) safety limit. In the Beni River, the mercury concentrations found in omnivorous and mud-feeding fish ranged from 0.02 to 0.19 microg g(-1) (wet wt.), and in piscivorous fish, from 0.33 to 2.30 microg Hg g(-1) (wet wt.). The mercury accumulated by carnivorous fishes was mainly present in its organic form; methylmercury represented 73-98% of the total mercury analysed. Eighty persons were studied in the entire Bolivian Amazonian basin. Unlike the gold miners, who are more affected by tropical diseases, such as malaria and yellow fever, the indigenous people living on the banks of the Beni river, present elevated levels of mercury (9.81 microg g(-1) on average). We observed an increase in contamination in young children still being breast-fed, confirming that hair mercury concentration in babies was significantly affected by maternal mercury contamination during pregnancy. These results show that the major health impacts caused by mercury affect people who are not working directly in gold mining activities but who have a regular fish diet. PMID- 11032118 TI - Elevated mercury concentrations in soils, sediments, water, and fish of the Madeira River basin, Brazilian Amazon: a function of natural enrichments? AB - Previous site-specific investigations have found that mercury concentrations in water, sediments, and biota of the Brazilian Amazon are elevated above global averages, and that these concentrations are a direct result of widespread mercury amalgamation mining operations conducted by non-organized prospectors. In order to assess the regional impacts of Hg contamination from these non-organized gold mining activities, water, sediments, and fish were systematically collected in 1997 along a 900-km reach of the Madeira River. The sampling program extended from the Amazon River upstream to Porto Velho, the site of historic and ongoing mercury amalgamation mining. Mercury concentrations were found to be elevated above global averages in all sampled media. However, the geochemical data suggest that the high mercury levels are due largely to natural sources and natural biogeochemical processes, and that the impacts of anthropogenically released mercury from mine sites is relatively localized. PMID- 11032120 TI - Three-dimensional modelling of mercury cycling in the Gulf of Trieste. AB - The Gulf of Trieste (Northern Adriatic) is subject to mercury pollution from a former mercury mine in Idrija, located along a river which transports mercury contaminated sediments into the Gulf. Concentrations in suspended and bottom sediments are up to two orders of magnitude higher than in the central and southern Adriatic. Extensive research has been carried out on measurements and modelling of the transport and fate of mercury in the Gulf. Two- and three dimensional models have been developed to include the influence of the significant advective transport due to currents. Wind, thermohaline forcing, and the Soca river momentum are the most important forcing factors. A two-dimensional model simulated the transport of non-methylated and methylated mercury in dissolved, particulate and plankton fractions. Mercury processes included the input of atmospheric mercury, sedimentation, reduction, methylation and demethylation. The model simulations gave basically what were proper trends of the phenomena; quantitatively the measured and computed results are mainly within a factor of three. To simulate the non-uniform distribution of parameters over the depth, an existing three-dimensional (3D) hydrodynamic and transport dispersion (TD) model, PCFLOW3D, was adapted and applied. As it was found that most mercury transport is related to suspended sediment particles, a new 3D sediment transport module was also developed and included in the model. Three cases are presented: one describing the simulation of TD of dissolved total mercury; another the simulation of the TD of particulate mercury in the Gulf during a river flood; and the third simulating sediment transport in the Gulf during a period of strong ENE wind. Comparison with measurements was only partly possible, but mainly the computed and measured results were within a factor of two and proper trends of the phenomena were obtained by the simulations. The combination of modelling and measurements has resulted in some interesting conclusions about the phenomenon of the transport and fate of mercury in a coastal sea. PMID- 11032119 TI - Relationship of mercury with aluminum, iron and manganese oxy-hydroxides in sediments from the Alto Pantanal, Brazil. AB - Sediments from nine floodplain lakes in Pantanal were analyzed for a large-scale (300 km) survey of mercury (Hg) load in sediments and soils of the Alto Pantanal and to study the relationship between Hg and reactive aluminum, iron, and manganese oxy-hydroxides. The results were compared with the Hg content in river and stream sediments from the Pocone gold mining area, where Hg has been extensively used and still is in use. The results indicate that the Hg concentrations were elevated in river sediment close to the mining area in Bento Gomes river basin (average in the < 74-microm fraction 88.9 ng Hg g(-1) dry wt.; interquartile range 50.3-119.5), but there was no clear indication that the local Hg emissions have contaminated the remote floodplain lakes, where concentrations were surprisingly low (average in the < 74-microm fraction 33.2 ng Hg g(-1) dry wt. sediment; interquartile range 18.4-46.8), in particular when considering geochemical characteristics of the sediment. The sediment from the floodplain lakes contained less Hg-tot and more reactive iron oxy-hydroxides than soils from the Tapajos area in the Amazon basin. This resulted in a mass ratio between Hg and amorphous oxy-hydroxides of only 5 x 10(-6) for Hg-tot/Fe-oxa (interquartile range 3-7 x 10(-6). PMID- 11032121 TI - Potential effects of forest fire and storm flow on total mercury and methylmercury in sediments of an arid-lands reservoir. AB - A study was conducted from July 1995 to June 1996 to examine the spatial and temporal changes of mercury concentrations in sediments of an arid-lands reservoir. Prior to the first sample collection in July, a forest fire burned 2930 ha of mixed conifer and ponderosa pine in the watershed of Caballo Reservoir in south-central New Mexico. The fire was eventually extinguished by summer rains and storm runoff resulting in the mobilization and transport of charred vegetative material into an intermittent tributary (Palomas Creek) that drains the watershed into Caballo Reservoir. Concentrations of total mercury (THg), monomethlymercury (MMHg), and total organic carbon (TOC) in surficial sediments revealed fire, followed by storm runoff, enhanced the transport of mercury and organic matter to the reservoir. Concentrations of THg in sediments increased from 7.5 etag/g in July to 46.1 etag/g by November 1995 at one site (Palomas) nearest the outflow of Palomas Creek. No other spatial or temporal trends were observed for THg at other sites throughout the remainder of the study. Concentrations of MMHg in sediments at the Palomas site increased from 0.428 etag/g in July to 12.46 etag/g by October 1995 compared to concentrations in sediments at the remaining sites which ranged from 0.11 to 1.50 etag/g throughout the study. The ratio of MMHg to THg (a gross index of methylation activity) was greatest in sediments from the Palomas site (5.4-33.8%) compared to the remaining sites (0.01-3.60%). The ratio was mirrored by elevated TOC in sediments at the Palomas site (2.5-11.8%) that remained elevated throughout the study. Fire and subsequent late-summer rains may have had a twofold effect on mercury concentrations in Caballo Reservoir. The storm-driven runoff following the forest fire carried mercury complexed to organic matter which resulted in elevated levels of mercury as well as providing a carbon source for microbial methylation processes in sediment. PMID- 11032122 TI - Export of mercury downstream from reservoirs. AB - Environmental effects monitoring at the La Grande hydroelectric complex (Quebec, Canada) revealed important increases in mercury levels in fish caught downstream from reservoirs. A study was carried out in 1997 immediately below the Caniapiscau Reservoir to identify by which components mercury is transported downstream from reservoirs and to assess the amount of mercury being exported. Stomach contents of lake whitefish (Coregonus clupeaformis) captured immediately below the Caniapiscau Reservoir were examined to determine which components transfer mercury from lower trophic levels to fish. The analyses of water samples and drifting organisms collected below the reservoir indicate that the dissolved fraction (< 0.45 microm) and the suspended particulate matter (0.45-50 microm) are the major components by which methylmercury is transferred downstream of reservoirs, accounting for 64 and 33%, respectively, of the total amounts exported. Drifting organisms such as plant debris, benthic invertebrates, fish, phytoplankton and zooplankton are much less important pathways for mercury export because of their very low biomass per water volume coming out of the generating station, as opposed to the high biomass of suspended particulate matter. However, zooplankton is the major component by which methylmercury is directly transferred to non-piscivorous fish downstream. PMID- 11032123 TI - Temporal and spatial influences of flooding on dissolved mercury in boreal reservoirs. AB - Dissolved total mercury and methylmercury concentrations ([HgT]D and [MeHg]D, respectively) in natural lakes and reservoirs of varying age (2 months to 69 years) were monitored between 1993 and 1997, during different times of the year and at several sites within the same reservoir. Both HgT-D and MeHgD in flooded environments were significantly greater (Student's t-test, P < 0.001) than those determined for neighbouring lakes, with average concentrations of 2.35+/-0.85 ng( 1) (n = 79) vs. 1.51+/-0.43 ng l(-1) (n = 70) for HgT-D and 0.28+/-0.10 ng Hg l( 1) (n = 87) vs. 0.05+/-0.025 ng Hg l(-1) (n = 30) for MeHgD. Furthermore, the proportion of HgT-D in the methylated form was, on average, nearly four times greater in the reservoir sites as compared with those of the lakes (12% vs. 3.6%). Simple linear regressions used to compare [MeHg]D with nine water quality parameters suggest that factors controlling the presence of MeHgD are not the same in all flooded environments. Overall, our results demonstrate that [MeHg]D are higher in reservoirs as compared with natural lakes and that up to 18 years after reservoir construction [MeHg]D remain elevated as compared with background levels. PMID- 11032124 TI - Discharge of methylmercury-enriched hypolimnetic water from a stratified reservoir. AB - A study conducted from July 1995 to June 1996 examining spatial and temporal distribution of mercury (Hg) at the Caballo Reservoir, New Mexico, revealed that the highest levels of methylmercury (MMHg) occurred in both the inlet and the Rio Grande upstream of the reservoir. As a result, a second study was designed to identify possible sources of the elevated levels of MMHg, and to determine if water discharged from the Elephant Butte Reservoir upstream could be a primary source. In July 1996, as anoxia began to develop in the hypolimnion of the Elephant Butte Reservoir, surface water MMHg concentrations were below the MDL of 0.018 ng/l while water discharged into the tailrace was 0.149 ng/l MMHg. By September 1996, when the anoxic hypolimnion spanned 60% of the total reservoir depth, surface water MMHg was still below the MDL, while discharge water had increased to 1.144 ng/l MMHg. Following reservoir turnover in November 1996, surface water increased to 0.264 ng/l MMHg while discharge water decreased to 0.420 ng/l MMHg. By January 1997, MMHg in the tailrace decreased to pre stratification levels, and both surface water and discharge water reached similar MMHg levels until the onset of summer stratification in July 1997. This trend was repeated the following year when MMHg concentrations in the tailrace increased from 0.190 ng/l in August 1997 to 1.240 ng/l in September 1997. In addition, vertical profile sampling of the reservoir from August 1997 to September 1997 showed a buildup of MMHg in the anoxic hypolimnion which coincided with increasing levels of MMHg discharged into the tailrace. During the course of this study the single largest contribution of MMHg to the river below the reservoir was from water released through the dam during the fall months of the year. PMID- 11032125 TI - Tests of the fidelity of lake sediment core records of mercury deposition to known histories of mercury contamination. AB - There has been recent controversy over the discrimination between natural and anthropogenic loadings of mercury to lakes. Sediment core profiles have been interpreted as evidence that inputs to lakes have increased. Some investigators have argued, however, that mercury may be sufficiently mobile in sediments to generate profiles that are misinterpreted as historical records. This argument can be tested where the histories of inputs of mercury are known independently from other kinds of information. We have such cases in Canadian lakes and we have been able to assemble sediment core records for comparison with known source histories. Three cases are represented by Clay Lake in Ontario where the source was a chlor-alkali plant with a known history of mercury discharges, Giauque Lake in the Northwest Territories where mercury was used at a gold mine to extract gold from ore, and Stuart Lake in British Columbia where a mercury mine operated for a known period at Pinchi Lake, the lake immediately upstream from Stuart Lake. In these cases lake sediment cores were dated using lead-210 and cesium-137 and then slices were analysed for mercury. The histories of mercury deposition derived from the cores agreed well with the known histories of inputs. PMID- 11032126 TI - Some potential errors in the measurement of mercury gas exchange at the soil surface using a dynamic flux chamber. AB - A series of controlled environment experiments were conducted to examine the use of a dynamic flux chamber to measure soil emission and absorption of total gaseous mercury (TGM). Uncertainty about the appropriate airflow rates through the chamber and chamber exposure to ambient wind are shown to be major sources of potential error. Soil surface mercury flux measurements over a range of chamber airflow rates showed a positive linear relationship between flux rates and airflow rate through the chamber. Mercury flux measurements using the chamber in an environmental wind tunnel showed that exposure of the system to ambient winds decreased the measured flux rates by 40% at a wind speed of 1.0 m s(-1) and 90% at a wind speed of 2 m s(-1). Wind tunnel measurements also showed that the chamber footprint was limited to the area of soil inside the chamber and there is little uncertainty of the footprint size in dry soil. PMID- 11032128 TI - Mercury fluxes in a natural forested Amazonian catchment (Serra do Navio, Amapa State, Brazil). AB - Mercury (Hg total) fluxes were calculated for rainwater, throughfall and stream water in a small catchment located in the northeastern region of the Brazilian Amazon (Serra do Navio, Amapa State), whose upper part is covered by a natural rainforest and lower part was altered due to deforestation and activities related to manganese mining. The catchment area is 200 km from the nearest gold mining (garimpo). Minimum and maximum Hg concentrations were measured monthly from October 1996 to September 1997 and were 3.5-23.4 ng l(-1) for rainwater, 16.5 82.7 ng l(-1) for throughfall (March-August 1997) and 1.2-6.1 and 4.2-18.8 ng l( 1) for stream water, in natural and disturbed areas, respectively. In the natural area, the inputs were 18.2 microg m 2 year(-1) in rainwater and 72 microg m(-2) year(-1) in throughfall. This enrichment was attributed to dry deposition. The stream output of 2.9 microg m(-2) year(-1) indicates that Hg is being recycled within the forest as other chemical species or is being retained by the soil system, as confirmed by the cumulative Hg burden in the 0-10 cm surface layer, which was 36480 microg m(-2). When the disturbed area of the catchment was included, the stream output was 9.3 microg m(-2), clearly indicating the impact of the deforestation of the lower part of the basin on the release of mercury. The Hg burden in the disturbed area was 7560 microg m(-2) for the 0-10 cm surface layer. PMID- 11032127 TI - Some local environmental effects on mercury emission and absorption at a soil surface. AB - The purpose of this study was to quantify effects of some local environmental variables on the soil surface exchanges of total gaseous mercury (TGM), under controlled conditions. A dynamic flux chamber with a Tekran mercury analyzer was used to quantify TGM emissions from, and absorption by, a clean, fine sandy loam soil with very low mercury content inside of a building and in a low TGM environment, outside. Simultaneous measurements of mercury flux, air and soil temperatures, ozone concentration, humidity, soil moisture and solar radiation were made. Controlled applications of water were made to change soil H2O content and measure the response of mercury flux. Air-soil exchanges were highly dependent on soil temperature (r2 = 0.78) and the mercury concentration gradient between the TGM in the soil pores and the ambient TGM above the soil surface (r2 = 0.98 for absorption and r2 = 0.408 for emissions). Correlations with air temperature and ozone levels are explained by the relationships of these variables with soil temperature. No detectable correlation was found with solar radiation or humidity. Wet soil maintained higher rates of soil TGM emission and decreased soil absorption. Emissions increased with increasing soil H2O, peaked at approximately field capacity, and then decreased slightly until saturation. PMID- 11032129 TI - Pools and fluxes of mercury and methylmercury in two forested catchments in Germany. AB - Mercury (Hg) and methylmercury (CH3Hg+) are global pollutants, but little information is available on rates of atmospheric input, distribution and mobility in soils and catchments of central Europe. The objectives of this study were to investigate input and output fluxes of these compounds in a deciduous and a coniferous catchment in NE Bavaria (Germany), and to estimate pools and mobility of total Hg (Hgtot) and CH3Hg+ at the catchment scale. Bulk precipitation, throughfall, litterfall and runoff were collected biweekly from April 1998 to April 1999. Several soil profiles were sampled to estimate pools of Hg compounds in the catchments. In both catchments highest contents of Hgtot were found in the Oa layer of the forest floor (up to 500 ng g(-1)) and the soil storage of Hgtot calculated for a soil depth of 60 cm was approximately 890 g ha(-1) in the coniferous and 190 g ha(-1) in the deciduous catchment. Highest contents of CH3Hg+ in upland soils were observed in the Oi layer of the forest floor, and soil storage of CH3Hg+ was 4.35 g ha(-1) in the coniferous and 0.59 g ha(-1) in the deciduous catchment. The annual total deposition of Hgtot (total deposition not measured directly but calculated from throughfall and litterfall) was 541 mg ha(-1) year(-1) in the coniferous and 618 mg ha(-1) year(-1) in the deciduous catchment. Total deposition rates of CH3Hg+ were 3.5 and 2.6 mg ha(-1) year(-1). The contribution of litterfall to the total deposition of Hgtot and CH3Hg+ was 55% in the deciduous catchment. In the coniferous catchment, the contribution of litterfall to total deposition was only 29% for Hgtot, but 55% for CH3Hg+. By far the largest proportion of the deposited CH3Hg+ and Hgtot remained in the catchments (85% in the coniferous, 95% in the deciduous). As compared to remote Swedish catchments, deposition and output via runoff of Hgtot, were higher, but deposition and output of CH3Hg+ were lower in our catchments. In contrast to other studies, the annual budget revealed no differences in the mobility between the two species at the catchment scale. However, temporal patterns of the runoff fluxes and converse gradients of CH3Hg+ and Hgtot contents in the forest floor indicated differences in mobility on shorter time scales. PMID- 11032130 TI - Sensitivity analysis of mercury human exposure. AB - We present a comprehensive analysis of the sensitivity of mercury (Hg) human exposure to environmental variables using a multimedia model of the fate and transport of Hg in the environment. The results of the analysis show that the Hg dose is most sensitive to the lake pH, the burial rate of Hg adsorbed to sediments, and the chemical speciation of Hg emissions to the atmosphere. The lake pH has a strong non-linear effect on the methylation rate and bioaccumulation of Hg in fish. The burial of sediments is a major pathway for removing Hg from the lake cycling. The speciation of Hg emissions is important because Hg(II) is deposited much more rapidly than Hg(0). These results highlight the importance of key variables that should be investigated through well-designed field programs, so that we can minimize the overall uncertainties associated with the modeling of mercury fate and transport. PMID- 11032131 TI - Steady-state transfer and depletion kinetics of mercury from amalgam fillings. AB - In 29 volunteers with a low amalgam load, the number of amalgam-covered tooth surfaces and the occlusal area of the fillings were determined. Before and at select times after removal of all amalgams, concentrations of total mercury were measured by cold-vapor atomic absorption in plasma and erythrocytes as well as in urine together with the excretion rate. Absorbed daily doses were estimated from intraoral Hg emission by two separate methods. The transfer of Hg from the fillings via the oral cavity and blood to urinary excretion was evaluated according to the most representative combination of parameters. This consisted of occlusal area (1), absorbed dose (2), Hg concentration in plasma (3) and urinary excretion (4). Pairwise correlation coefficients were 0.49 for parameters 1 vs. 2, and 0.75 each for parameters 2 vs. 3 and 3 vs. 4. Within 9 days after removal of the fillings, a transient increase in Hg levels was observed in plasma only; in the group without a rubber dam, concentrations increased significantly above pre-removal values at days 1 and 3, whereas they decreased significantly below pre-removal values at day 30 in the rubber-dam group and at day 100 in both groups. Excretion rates decreased significantly at day 100 in the protected group. Peak plasma-Hg was 0.6 ng/ml on average at day 1 and decreased with halftimes of 3 and 43 days in subjects protected by a rubber dam. The results indicated that concentrations of total mercury in plasma responded rapidly to changes in the amalgam status and reflected the actual absorption most reliably. Notably, plasma-Hg levels were sensitive enough to detect a transient attenuation of the additional exposure after using a rubber dam during the removal of only a few fillings. However, being small in magnitude and lasting 100 days at best, the rubber-dam effect had minor toxicological relevance. PMID- 11032132 TI - Methylmercury and the health of indigenous peoples: a risk management challenge for physical and social sciences and for public health policy. AB - Methylmercury in aquatic ecosystems and bio-accumulated in aquatic biota, especially fish, is a major public health concern internationally. Precautionary efforts are currently underway internationally to reduce the anthropogenic release of mercury, which in turn, over time, will reduce human exposure. However, at the present time, it is important to address the issue of management of the risks of exposure as they exist now. Of particular concern are the impacts of methylmercury on indigenous populations which depend on fish as a subsistence food source, both in remote areas of developed countries, such as Canada, and in developing countries such as Brazil. Research into these impacts over the past two or three decades has shown that, other than in very severe pollution situations such as occurred in Minamata, Japan, the direct impacts on human health are difficult to prove. On the other hand, the indirect negative effects of methylmercury on health, mediated through the disruption of lifestyle and eating patterns and the associated socio-cultural and socio-economic consequences among the affected native populations, have, in many cases, been significant. These social factors have raised serious challenges in determining practical public health policies on the issue. Policy development relating to environmental contaminants has been presented, with the problem of assessing the role of the various factors which contribute to the impact on health as a result of socio cultural disruption. These factors include changes in diet and lifestyle due to methylmercury in the environment and its real or perceived risk. The standard physical sciences risk assessment process, based on the lowest observed adverse effects level (LOAEL) or no observed adverse effects level (NOAEL) used in defining health policies may be seen as over-simplistic theoretical extrapolations when viewed in the context of the concerns of the social sciences. Both approaches, however, have relevance to health policies that address the risks posed by environmental methylmercury. Therefore, the standard physical sciences approach of the past three decades now needs to be linked with the social sciences approach, with its focus on the indirect impacts of exposure to methylmercury, to provide a comprehensive approach to public health policy development. With this objective in mind, this paper reviews methylmercury related data from both physical and social sciences. It attempts to draw on the findings in both disciplines to provide suggestions for an integrated approach in policy development relating to human health and human exposure to methylmercury, especially among indigenous peoples in remote areas and in developing countries. An integrated approach such as this may help to limit adverse health effects in the indigenous communities affected. PMID- 11032133 TI - Health assessment for mercury exposure among schoolchildren residing near a gold processing and refining plant in Apokon, Tagum, Davao del Norte, Philippines. AB - Artisanal gold-mining activities in the Philippines have proliferated since the early 1980s. Presently, environmental and health monitoring conducted by several governmental agencies is limited to the determination of total mercury only. Previous studies undertaken focused mainly on the exposure of adults and workers to mercury during mining/processing operations. However, in one area in Mindanao, mined ores are brought down and processed in the lowlands where residential communities are exposed to environmental pollutants resulting from gold processing/refining operations. The area of study is Apokon, Tagum, Davao del Norte, which has 29 gold processing and refining plants. Health complaints among schoolchildren in Apokon Elementary School were received by the Department of Health and were attributed to the mercury pollution in the environment. As part of a collaboration with the Health Department, UP-National Poisons Control and Information Service, the National Institute for Minamata Disease (NIMD), Japan, provided technical assistance in the analytical determination of mercury in biological and environmental samples. Elevated mercury concentrations were noted in some of the river systems up to 15 km from the mining areas. Environmental quality monitoring showed T-Hg sediment levels ranged from 0.553 to 66.471 microg/g dry wt. while water samples from river systems exhibited mercury levels from 72.8 to 78.4 ng/ml. Twenty-seven sediment samples from river systems near mining operations and seven water samples were also brought to the Institute for analysis. Fish samples collected showed levels ranging from 1.07 to 438.8 ng/g for total mercury and 0.71-377.18 ng/g for methylmercury. Methylmercury content in fish is predominant. All water and sediment samples collected from three sampling sites have elevated T-Hg level while three fish species have elevated T Hg and methylmercury levels (WHO/CDC, 1994). Blood and hair samples from 162 schoolchildren aged 5-17 years were collected and analyzed at the NIMD for mercury analysis. Analytical procedures used in the NIMD for mercury testing were applied. Laboratory results showed that total mercury hair samples ranged from 0.278 to 20.393 microg/g while methylmercury hair results were from 0.191 to 18.469 microg/g. Methylmercury in hair showed levels from 45.96 to 99.81%. Total blood mercury levels ranged from 0.757 to 56.88 microg/l while Me-Hg blood levels ranged from 1.36 to 46.73 microg/l. It was determined that 10 children had elevated T-Hg blood levels while one child had high total and methylmercury levels in hair. A summary of physical examination results showed that the predominant findings include under-height, gingival discoloration, adenopathy, underweight and dermatologic abnormalities among children examined. PMID- 11032134 TI - Human exposure to mercury due to small scale gold mining in northern Tanzania. AB - In northern Tanzania large numbers of small scale miners use mercury in the gold extraction process causing contamination of the environment and risks to human health. Human exposure to Hg was assessed in populations in and around small scale gold mining camps by means of human hair and urine surveys. We also determined Hg concentration in fish in aquatic bodies close to these camps. Urinary Hg testing in three communities showed that 36% of the gold miners working with amalgam exceeded the WHO guideline concentration of 50 microg Hg/g creatinine. Data from a hair survey of fishermen and farmers confirm that at present, the fish-eating population close to the southern tip of Lake Victoria is at low risk with regard to Hg exposure. Concentrations in fish were low and > 90% of the hair samples from the fish-eating population were below 2 microg/g T-Hg. Highest Hg concentrations in fish caught along the southern shores of Lake Victoria and in rivers draining from gold processing sites were detected in lungfish species (Protopterus aethiopicus), and lowest Hg concentrations in tilapia (Oreochromis niloticus and Tilapia zilii). PMID- 11032136 TI - Assessing the contribution of natural sources to regional atmospheric mercury budgets AB - Naturally mercury-enriched substrate is a long-lived source of mercury to the global atmospheric mercury cycle. Field flux chambers, laboratory gas exchange chambers and micrometeorological methods may be applied to estimate emissions from these sources. However, field chamber experimental design may affect the magnitude of the fluxes measured, and the laboratory chamber only provides a minimum estimate of flux. Many factors, such as mercury concentration and speciation in substrate, light, precipitation, and temperature, influence the emission of mercury from the substrate. Mercury concentration in the substrate is a dominant factor controlling emissions and may be used to predict emissions from regions of mercury enrichment. Mercury fluxes measured from three areas of natural enrichment and three areas with low levels of mercury enrichment are 1-5 orders of magnitude greater than the value applied to global belts of natural enrichment. Preliminary scaling of emissions from one of these areas and for western North America indicates that mercury enriched areas may be significant sources of mercury to the atmosphere, and that their contribution to regional and global atmospheric budgets needs to be reassessed. PMID- 11032135 TI - Some results on Hg content in hair in different populations in Albania. AB - The mercury content in hair, for exposed dental workers and unexposed groups of people living in Albania was studied. The influence of factors such as age, sex, body weight, fish consumption in the diet, number of dental amalgam fillings and time exposed to mercury, was determined. The mean value of mercury content in hair for the subjects under study (0.705 microg/g) was found to be lower than the value referred from the World Health Organization (WHO) for people who do not consume fish with a high methylmercury content (1-2 microg/g). A significant positive correlation of Hg content with the number of dental amalgam fillings, time under exposure and frequency of fish consumption in the diet was found. These correlations became stronger when exposed and unexposed groups of people were evaluated separately. PMID- 11032137 TI - Assessment of atmospheric mercury emissions in Finland AB - This paper is part of the study of atmospheric emissions of heavy metals conducted by the Finnish Environment Institute in collaboration with the Technical Research Centre of Finland (VTT) under the umbrella of the Finnish Ministry of the Environment. The scope of our study is limited solely to anthropogenic mercury that is emitted directly to the atmosphere. This article addresses emission factors and trends of atmospheric mercury emissions during the 1990s and is based mainly on the database of the Finnish Environmental Administration. In addition, data based on the measurements taken by the VTT regarding emission factors have been used to estimate emissions of mercury from the incineration of waste. The study indicates that the total emission of mercury has decreased from 1140 kg in 1990 to 620 kg in 1997, while industrial and energy production have been on the increase simultaneously. The 45% emission reduction is due to improved gas cleaning equipment, process changes, automation, the installation of flue gas desulfurization process in coal-fired power plants and strict pollution control laws. In the past, some authors have estimated a higher mercury emission in Finland. In this study, it is also observed that there are no big changes in the quality of raw materials. Estimated emission factors can be of great help to management for estimating mercury emissions and also its risk assessment. PMID- 11032138 TI - Influence of the Sostanj coal-fired thermal power plant on mercury and methyl mercury concentrations in Lake Velenje, Slovenia AB - Lake Velenje is located in one of the most polluted regions in Slovenia, the Salek Valley. The major source of pollution in the valley is the coal-fired thermal power plant in Sostanj (STPP, capacity 775 MW). It has five separate units. All units have electrostatic precipitators for fly ash removal. Unit 4 also has installed a wet flue gas desulfurisation system (FGD system). Total mercury (THg) concentrations were measured in lignite, slag and ash samples from the STPP. In flue gas, different mercury species (THg, MeHg, Hg2+, Hg0) were determined separately for unit 4 and unit 5 which use different flue gas cleaning technology. Mercury and methyl mercury (MeHg) concentrations were also measured in lake water at different depths, in inflow water, outflow water, rain, snow and lake sediments in order to establish the influence of the power plant on the lake. Most mercury emitted from the power plant is in the elemental form. The ratio between oxidised and elemental Hg depends on the flue gas cleaning technology. Mass balance calculations have been performed for the STPP. The results show that the major sources of mercury in Lake Velenje are wet deposition and lake inflows. Total and MeHg concentrations in the water column are very low and can be compared to other non-contaminated freshwater lakes in the world. PMID- 11032139 TI - Mercury emissions from a coal-fired power plant in Japan AB - The emissions study for mercury was conducted at a 700 MW coal-fired plant for the combustion of three types of coal with mercury concentrations of 0.0063, 0.0367 and 0.065 mg/kg. The power plant is equipped with a cold-side electrostatic precipitator and wet type flue gas desulfurization system. During full load operation of the boilers, samples of the input and output streams such as coal, coal ash, ESP ash and post-ESP particulates and flue gas were collected. The Hg concentrations in solid were measured by cold-vapor atomic absorption spectrometry (AAS) after appropriate preparation and acid digestion. Gaseous Hg was collected using a mixed solution of potassium permanganate and sulfuric acid and the Hg concentrations in the samples were measured using cold-vapor AAS. The results were used to examine: (1) overall mass balances; (2) relative distribution in the power plant; (3) equilibrium of Hg species using MALT-2 calculation program; and (4) Hg concentrations in stack emissions. The mass balances estimated in this study were 100, 138 and 89%, respectively, for the coals. Total Hg concentrations in stack gas were 1.113, 0.422 and 0.712 microg(m3N), respectively, for the coals. More than 99.5% of the Hg in the stack emissions were in gaseous form and the proportion in particulate form was extremely low. The relative distribution of Hg in ESP, FGD and Stack ranged from 8.3 to 55.2%, 13.3 to 69.2% and 12.2% to 44.4%, respectively. The results indicated that factors other than the Hg concentration of coals and efficiency of pollution control devices might affect Hg emissions from coal-fired plant. The calculated equilibrium of the distribution of Hg species using the MALT2 program suggest that it is necessary to consider condensation mechanism to interpret the affect of Hg species on the variations of the removal efficiencies of Hg in the ESP. PMID- 11032140 TI - Mercury contamination associated with small-scale gold mining in Tanzania and Zimbabwe. AB - Mercury contamination associated with small-scale gold mining and processing represents a major environmental and human health concern in Eastern and Southern Africa. Approximately 200,000-300,000 persons are involved in small-scale gold mining activities in Tanzania and > 200,000 persons in Zimbabwe. Mercury (Hg) is used mainly for the processing of primary gold quartz veins and supergene gold mineralizations. Gravimetric material flow analyses show that 70-80% of the Hg is lost to the atmosphere during processing, 20-30% are lost to tailings, soils, stream sediments and water. For every 1 g Au produced, 1.2-1.5 g Hg are lost to the environment. Cumulatively, the anthropogenic Hg released annually into the atmosphere is approximately 3-4 t in the whole Lake Victoria Goldfields of Tanzania and > 3 t in Zimbabwe. Tailings are local 'hot spots' with high concentrations of As, Pb, Cu and Hg. Lateral and vertical dispersion of Hg lost to soils and stream sediments is very limited (laterally < 260 m, vertically < 20 cm). Dispersion of mercury from tailings is low because Hg is transported largely in the elemental, metallic form. In addition, Fe-oxide rich laterites and swamps appear to be natural barriers for the dispersion of metals in soils and streams. Ground and surface water quality data indicate very low dispersion rates during the dry season. PMID- 11032141 TI - Volcanoes as emission sources of atmospheric mercury in the Mediterranean basin AB - Emissions from volcanoes, fumaroles and solfataras as well as contributions from widespread geological anomalies could represent an important source of mercury released to the atmosphere in the Mediterranean basin. Volcanoes located in this area (Etna, Stromboli and Vulcano) are the most active in Europe; therefore, it is extremely important to know their mercury contributions to the regional atmospheric budget. Two main methods are used for the evaluation of volcanic mercury flux: a direct determination of the flux (by measuring in the plume) and an indirect one derived from the determination of the Hg/SO2 (or Hg/S) ratio value, as SO2 emissions are constantly monitored by volcanologists. An attempt to estimate mercury flux from the Vulcano volcano and to establish the Hg/S ratio value has been made along three field campaigns carried out in October 1998, in February and May 1999 sampling several fumaroles. Traditional sampling methods were used to collect both total Hg and S. The average Hg/S ratio value resulted to be 1.2 x 10(-7). From the Hg/S value we derived the Hg/SO2 value, and by assuming that all the volcanoes located in this area have the same Hg/SO2 ratio, mercury emissions from Vulcano and Stromboli were estimated to be in the range 1.3-5.5 kg/year and 7.3-76.6 kg/year respectively, while for Etna mercury flux ranged from 61.8 to 536.5 kg/year. Data reported in literature appear to be overestimated (Fitzgerald WF. Mercury emission from volcanos. In: 4th International conference on mercury as a global pollutant, August 4-8 1996, Hamburg, Germany), volcanic mercury emission does not constitute the main natural source of the metal. PMID- 11032142 TI - Air/water exchange of mercury in the Everglades I: the behavior of dissolved gaseous mercury in the Everglades Nutrient Removal Project AB - From 1996 to 1998 we determined dissolved gaseous mercury (DGM) in waters of the Everglades Nutrient Removal Project (ENR), a constructed wetlands. The concentrations of DGM measured in these waters (mean 7.3 +/- 9.5 pg l(-1)) are among the lowest reported in the literature, and suggest a system often near or slightly above equilibrium with Hg in ambient air. DGM exhibited both seasonal and diel trends, peaking at midday and during the summer. A simple box budget model of DGM in waters of the Everglades was developed using an interactive spreadsheet based on a mass balance among light-induced reduction of HgII (production of DGM), Hg0 oxidation (removal), and Hg0 evasion in a box (water column) consisting of a surface region with sunlight available and a lower dark region. The modeling results suggest high sensitivity of hourly DGM concentrations to DGM production rates and initial DGM levels. The sensitivity to Hg oxidation is lower than the sensitivity to DGM production. The model performance demonstrates successful simulations of a variety of DGM trends in the Everglades. In particular, it clearly demonstrates how it is possible to measure comparable rates of evasion over several Everglades sites with different DGM concentrations. PMID- 11032143 TI - Air/water exchange of mercury in the Everglades II: measuring and modeling evasion of mercury from surface waters in the Everglades Nutrient Removal Project AB - During the period 1996-1998, and covering each of the four climatic seasons, we measured evasion of Hg using flux chambers over waters of the Everglades Nutrient Removal Project (ENR), a constructed wetlands. In situ water evasion fluxes of total gaseous mercury (primarily Hg0) measured at seven Everglades sites typically show low evasion rates (< 3 ng m(-2) h(-1)), which are smaller on average than those measured in northern systems. Evasion exhibits a strong diel cycle, with the highest values during midday, and evasion rates are correlated with dissolved gaseous mercury (DGM), solar radiation, wind speed, and water temperature. A thin film model was adapted and applied to estimate evasion fluxes for these Everglades waters based on our measurements of DGM. The results suggest that the typical thin film model underestimates evasion for the Everglades waters, at least on a short time scale. PMID- 11032144 TI - Modeling assessment of transport and deposition patterns of anthropogenic mercury air emissions in the United States and Canada AB - In December 1997, the U.S. Environmental Protection Agency submitted its mercury study report to Congress, which included a modeling assessment of the long-range transport and deposition of mercury from various residential and industrial sources within the United States, based on numerical simulations of elemental mercury gas, divalent mercury gas and particulate mercury, using a special version of the Regional Lagrangian Model of Air Pollution (RELMAP). Observations of the deposition of total precipitated mercury at several locations within the U.S. were compared to the modeling results in order to evaluate the accuracy of the RELMAP simulations. However, the lack of Canadian mercury emissions data in the RELMAP modeling limited the usefulness of this model evaluation at locations near the Canadian border. An inventory of Canadian mercury emissions obtained from Environment Canada has now been added to the RELMAP mercury modeling, and the previous simulations and evaluation of modeled wet deposition have been repeated. The results indicate that emissions of mercury from Canada, as represented by the new inventory, do not significantly impact simulated wet deposition over the United States, nor the general results of the previous model evaluation. Analyses of the simulated transport and deposition patterns of mercury emissions from both the United States and Canada are presented, along with the simulated fraction of total mercury wet deposition, separately attributable to sources within each nation. PMID- 11032145 TI - On artificial dilution of point source mercury emissions in a regional atmospheric model AB - Previously, we developed and applied a regional atmospheric mercury model to a domain covering most of North America at a horizontal grid resolution of 100 km. The implication of using this coarse resolution is that point sources of mercury emissions are instantaneously spread over a grid volume of horizontal dimensions 100 x 100 km2 and a vertical dimension equal to the depth of the grid cell where the point source emissions are released. Since point sources comprise a significant majority of a regional mercury emissions inventory, it is important to understand what effect this artificial dilution may have on calculated mercury concentrations and deposition fluxes. To understand this effect, we conducted model simulations using a finer grid, embedded within the original coarse grid, over a sub-domain that includes over 50% of the largest mercury point sources in the north-eastern United States. The horizontal resolution of the fine grid is 20 km, i.e. it is five times smaller than that of the coarse grid. We compared short term (daily) and long-term (annual) averaged mercury concentrations, and deposition (wet and dry) fluxes on the coarse and fine grids. As expected, the effect of grid resolution is more clearly seen in close proximity to point sources than at remote locations. For short-term averages near major point sources, the peak concentrations and dry deposition fluxes of mercury from the fine grid are almost a factor of two greater than the corresponding estimates from the coarse grid. At remote locations, however, the concentrations and dry deposition peaks estimated by the two model grid resolutions are more comparable. For total wet deposition of mercury, the distinction between the fine and the coarse grid model results is less significant, regardless of the location. This could be due to the redistribution of precipitation fields or the effect of mercury aqueous chemistry. The effect of grid resolution is more important when model estimates are averaged over short time periods, e.g. daily, as opposed to over long periods, e.g. seasonally and annually. PMID- 11032146 TI - A sensitivity analysis on the atmospheric transformation and deposition of mercury in north-eastern USA AB - This paper presents the results of a sensitivity analysis on the factors that affect dry and wet deposition of atmospheric mercury (Hg), using a regional scale air quality model. Simulations were conducted for the north-eastern USA during a summer week and a winter week in 1997. Simulation results for the summer week and the winter week in general showed similar responses to changes in emission, environmental conditions, and alternative chemical mechanisms. Reduction of the ambient concentrations of soot or ozone was shown to reduce the wet deposition of Hg. When averaged over the summer and the winter week, the total deposition to the simulation domain would be reduced by 26% by reducing Hg emission from anthropogenic sources within the domain by 50%. For individual grids, however, only locations near local sources obtained noticeable reductions in ambient concentration and wet deposition due to the influence of re-emission from the natural surfaces and regional/global scale transport. The reduction in deposition would reach 36% if all Hg(II) emitted from anthropogenic sources were attached to particles. The total deposition was predicted to decrease by 22% when the gas phase Hg(II)-Hg(p) partitioning was included in the model. Only small changes in total deposition were observed by including the gas-phase ozone-Hg(0), reaction and the aqueous phase chlorine-Hg(0), reaction, and by lowering ambient concentrations of Hg(II) and Hg(p) at the upper lateral boundaries. During the summer week, Hg(II) deposition contributed 40% or more to the total deposition. The contribution increased to 70% in the winter week. PMID- 11032147 TI - Temporal trends in gaseous mercury evasion from the Mediterranean seawaters. AB - Mercury evasion from seawaters is considered to be one of the main natural sources of mercury released to the atmosphere. The temporal evolution of this mechanism is related to biotic and abiotic processes that produce mercury in its elemental form and as DGM. The efficiency of these processes depends upon the intensity of the solar radiation, the ambient temperature of the air parcel above the seawater, and the water temperature. In the Mediterranean region, the magnitude of these mechanisms are particularly significant, due to favorable climate conditions and to the presence of large cinnabar deposits that cross the whole region; all these synergic factors yield significant evasional fluxes of mercury from the surface water during most of the annual period. In this work, mercury fluxes were measured by using a floating flux chamber connected to an atomic absorption analyzer. Photosynthetic active radiation (PAR) and UV components of the solar radiation were measured using the same system adopted in the EC 'ELDONet project'. The measurements of the mercury evasional fluxes were carried out at three sites of the northern Tyrrhenian Sea during 1998. Two sites were located at unpolluted and polluted coastal areas, and the third was an offshore site. The evasional flux showed a typical daily trend, highest at midday when the ambient temperature and solar radiation were at the maximum, and lowest, near to zero, during the night. Besides the day-night behavior, a seasonal trend was also observed, with minimum values during the winter period (0.7-2.0 ng/m2 h) and maximum values during the summer (10-13 ng/m2 h). PMID- 11032148 TI - Total gaseous mercury in Quebec (Canada) in 1998 AB - The main objectives of this paper were to report total gaseous mercury (TGM) concentrations at four stations along the St. Lawrence River in 1998 and to give some insights regarding their fate. The TGM network stretches from latitudes 45 degrees N to 50 degrees N. TGM and ozone concentrations (used as complementary pollutant) were presented. The median TGM concentrations throughout the network varied from 1.62 to 1.79 ng/m3. Sites close to Montreal had larger TGM concentrations. Montreal surroundings appeared to be a source of TGM. The TGM concentrations were maximum in wintertime and minimum in summertime throughout the sites. However, the diurnal variations were site specific. St. Anicet and L'Assomption were more variable in terms of large concentrations, whereas Mingan and Villeroy were more variable in terms of low concentrations. Ozone and TGM concentrations were positively correlated at some sites (Villeroy, Mingan and St. Anicet) whereas they were not at L'Assomption (closest site to Montreal). Local TGM and ozone precursor sources would affect the correlation. PMID- 11032149 TI - Gas-phase mercury in the atmosphere over the southern Baltic Sea coast AB - Results of atmospheric total gaseous mercury (TGM) measurements performed at two Baltic Sea coastal stations, Peninsula Hel (Poland) and Preila (Lithuania), from June 16 to August 11, 1997, are presented. High time-resolution data were obtained by using automated atomic absorption mercury vapor analyzers (Model Gardis-1A). Analysis of TGM concentration data (directional distribution, correlation with meteorological parameters, diurnal variability) detected the Baltic sea, in particular its southern part and Gulf of Gdansk, as the main gaseous mercury source for the region during the summer months. The source seemed to be activated by solar radiation, air temperature, and, probably, wind. PMID- 11032150 TI - Elemental gas phase atmospheric mercury as it interacts with the ambient aerosol and its subsequent speciation and deposition AB - A model describing the interaction of gas phase elemental atmospheric mercury, H(g)(o), with the ambient aerosol has been linked to hybrid single-particle Lagrangian integrated trajectory (HYSPLIT_4) a hybrid dispersion and deposition model. The integrated modelling system allows H(g)(o) emitted from an anthropogenic source in the presence(g) of the local aerosol to be followed and calculates the deposition fluxes to terrestrial and water receptors resulting from the interaction between the H(g)(o) and the aerosol. The model includes a source-based parameterized description of the ambient aerosol, a gas-phase diffusion and adsorption model for the interaction of H(g)(o) and the ambient particulate matter under conditions of low relative humidity, and an aqueous phase Hg chemistry model to allow for complete or partial deliquescence of the particles under conditions of high humidity. The model is, thus, able not only to calculate deposition fluxes, but also the speciation of the Hg compounds deposited; this is of great importance in the modelling of the atmospheric Hg cycle due to the difference in the properties of elemental and oxidized Hg. PMID- 11032151 TI - Atmospheric mercury deposition in Guizhou, China AB - The atmospheric deposition of mercury including total gas phase mercury (TGM), gas phase divalent mercury (Hg2+), and gas phase monomethyl mercury (MMHg) was investigated. Samples were collected from central cities and nature reserve areas across the province. Techniques and equipment including moss bags, KCl-coated denuder and GARDIS-1A portable mercury vapor analyzer were employed for sample collection and analysis. The reliability of techniques and analytical methods used for the project was evaluated. The deposition of atmospheric mercury was reported in species in this important mercury polluted province for the first time. The spatial and temporal variability of mercury emissions were investigated and reported. Contribution of coal mercury emissions to the atmosphere mercury deposition of the province was also investigated. PMID- 11032152 TI - Background levels of atmospheric mercury in Kagoshima City, and influence of mercury emission from Sakurajima Volcano, Southern Kyushu, Japan AB - Vapor phase mercury concentration was determined daily for 1 year (Jan. 1996-Jan. 1997) in order to present the levels of atmospheric mercury in Kagoshima City and to estimate the influence of mercury emission from Sakurajima Volcano, southern Kyushu, Japan. The atmospheric mercury was collected on a porous gold collector at Kagoshima University and was determined by cold vapor atomic absorption spectrometry; Kagoshima University of Kagoshima City is located approximately 11 km west of Sakurajima Volcano. The mercury concentration obtained was in the range 1.2-52.5 ng m(-3) (mean 10.8 ng m(-3), n = 169). The atmospheric concentration varied from season to season; the concentration was high in summer and lower in winter. A linear relation was obtained by plotting ln[Hg/ng m(-3)] vs. 1/T for the north, south and west winds with correlation coefficients of 0.76, -0.79 and -0.83, respectively, but no such dependency was found for the east wind (r = -0.035). When the wind is blowing from the east, Kagoshima City is on the leeward side of the volcano. The impact of the fumarolic activity of the volcano on ambient air in the city was evident in the disappearance of temperature dependency with the appearance of the east wind. Atmospheric mercury concentration except for the east wind was considered to be background levels of Kagoshima City. As background levels, 8.1 +/- 5.3 ng m(-3), 14.8 +/- 7.9 ng m( 3), 13.9 +/- 11.7 ng m(-3) and 4.4 +/- 1.6 ng m(-3) (mean +/- S.D.) were obtained for spring, summer, autumn and winter, respectively. PMID- 11032153 TI - Gestational age and growth rate of fetal mass are inversely associated with systolic blood pressure in young adults: an epidemiologic study of 165,136 Swedish men aged 18 years. AB - The objective of this study was to identify specific components of fetal growth that may underlie the observed association between birth weight and later blood pressure. A record linkage was made between the Swedish Medical Birth Registry, the Military Conscription Register, and censuses. For 165,136 men born in Sweden between 1973 and 1976 and conscripted from 1990 to 1996, systolic blood pressure was measured at age 18 years. Systolic pressure was independently inversely associated with birth weight for gestational age and with gestational age itself but not with birth length for gestational age. The difference in systolic pressure between the top and the bottom quintiles of birth weight for gestational age was -1.61 mmHg (95% confidence interval: -1.82 to -1.40) after adjustment for birth length for gestational age, height, and weight. The change in systolic pressure was -0.25 mmHg (-0.29 to -0.22) for a 1-week increase in gestational age. How far the inverse association of systolic pressure with length of gestation represents an independent effect of maturation is unclear. These findings help to refine the fetal origins hypothesis and provide further criteria against which potential biological mechanisms that link circumstances in utero to later blood pressure can be assessed. PMID- 11032154 TI - Invited commentary: association between restricted fetal growth and adult chronic disease: is it causal? Is it important? PMID- 11032155 TI - Poor agreement between self-reported birth weight and birth weight from original records in adult women. AB - Data from an ongoing prospective population study of women in Goteborg, Sweden, were used to assess agreement between self-reported birth weight and birth weight obtained from original delivery records of women aged 44-60 years. Of the eligible population with traced delivery records (n = 693), only 28% (n = 192) could report their own birth weight. Spearman correlation between self-reported birth weight and birth weight from original records was r = 0.76. However, a difference plot, with limits of agreement at -1,028 to 1,038 g (95% confidence limits: lower limit, -1,157 to -901 g, upper limit, 910 to 1,166 g) revealed poor agreement between methods. Of the self-reported birth weights, 53% were in error by 250 g or more, and 31% were positively or negatively discordant by 500 g or more. Application in an analysis of cardiovascular risk factors in adulthood found conflicting results between self-reported and recorded birth weights. Low reporting rate, poor reporting accuracy, and misleading findings in application led to the conclusion that self-reported birth weights from middle-aged women would not be a satisfactory replacement for birth weights from original records. PMID- 11032156 TI - Association of educational achievement with pulsatile arterial diameter change of the common carotid artery: the Atherosclerosis Risk in Communities (ARIC) Study, 1987-1992. AB - Education is strongly inversely associated with common carotid artery intima media thickness in the Atherosclerosis Risk in Communities (ARIC) Study. The authors extended the ARIC study of preclinical atherosclerosis by evaluating the cross-sectional association of education with common carotid artery elasticity. This study included 10,091 Black and White men and women aged 45-64 years who were free of clinical coronary heart disease and stroke/transient ischemic attack. Arterial elasticity was assessed by pulsatile arterial diameter change (PADC), derived from phase-locked echo-tracking. The smaller the PADC, the stiffer the artery. Education was categorized into grade school, high school without graduation, high school with graduation, vocational school, some college, and graduate/professional school. PADC was directly associated with educational attainment. The mean PADCs, adjusted for age, height, diastolic diameter, systolic blood pressure, pulse pressure (linear and squared), ethnicity, gender, and smoking status, in successively higher education strata were 402 (standard error (SE) 5), 403 (SE 4), 407 (SE 3), 413 (SE 4), 416 (SE 2), and 417 (SE 4) microm (p = 0.007). To the authors' knowledge, this is the first time such an association has been reported. If arterial dilation impairment precedes arterial wall thickening in the atherosclerotic process, as recent studies on endothelial dysfunction suggest, these results indicate that low socioeconomic status may be associated with early arterial pathophysiologic changes-an effect that appears to be mediated by established cardiovascular disease risk factors. PMID- 11032157 TI - Factors predictive of ischemic heart disease mortality in foundry workers exposed to carbon monoxide. AB - The potential predictors of ischemic heart disease mortality were studied for 931 male foundry workers in Finland who participated in a health examination in 1973. These workers were followed up to 1993 through registers and by using a questionnaire. In 1973, the systolic and diastolic blood pressures of workers exposed to carbon monoxide (CO) were slightly higher than those of unexposed workers. The prevalence of angina pectoris showed a clear dose-response relation to CO exposure. Electrocardiogram (ECG) findings indicating past myocardial infarction or suggesting coronary artery disease as a function of smoking and/or CO exposure were not evident. In the 1987 follow-up, the rate ratio for ischemic heart disease mortality was estimated as 4.4 for CO-exposed smokers compared with unexposed nonsmokers. Ischemic heart disease mortality in 1973-1993 was analyzed by using the Cox proportional hazards model. The statistically significant predictors were age, pathologic ECG findings in 1973, regular CO exposure, and abundant alcohol drinking. Of the ECG findings, changes in Q or QS and ST-J or ST waves and in ventricular extrasystoles were statistically significant. The risk of mortality from ischemic heart disease was increased by working in iron foundries, by hypertension, and by smoking. PMID- 11032158 TI - Visual impairment and risk of hip fracture. AB - As part of a case-control study, the Auckland Hip Fracture Study (1991-1994), the authors examined associations between impaired vision and risk of hip fracture. Subjects (911 cases and 910 controls aged 60 years or older) completed a questionnaire and had vision measurements taken, including measurements of visual acuity and stereopsis (depth perception). Binocular visual acuity worse than 20/60 was statistically significantly associated with increased risk of hip fracture after adjustment for age, sex, proxy response, hours of activity per week, and height (odds ratio (OR) = 1.5; 95% confidence interval (CI): 1.1, 2.0), as was having poor vision (less than 20/100) in both eyes (OR = 2.4; 95% CI: 1.0, 6.1). Having no depth perception was associated with increased risk (OR = 6.0 95% CI: 3.2, 11.1), as were categories of decreasing stereopsis (trend p = 0.0001), self-reported poor vision (OR = 1.4; 95% CI: 1.0, 1.9), not wearing glasses at the time of the fall (OR = 1.2; 95% CI: 1.0, 1.6), and increasing time since the last eye examination (trend p = 0.03). The population attributable risk of hip fracture due to poor visual acuity or stereopsis was 40%. Visual factors are important fall-related factors which influence risk of hip fracture. Risk of hip fracture may be decreased by correcting refractive error, improving stereopsis, and administering regular eye examinations. PMID- 11032159 TI - Increased risk of agricultural injury among African-American farm workers from Alabama and Mississippi. AB - Research on the epidemiology of agriculture-related injuries has largely ignored African-Americans and farm workers. This cohort study is the first to estimate injury rates and to evaluate prospectively risk factors for agriculture-related injuries and compare them among African-American and Caucasian farmers and African-American farm workers. A total of 1,246 subjects (685 Caucasian owners, 321 African-American owners, and 240 African-American workers) from Alabama and Mississippi were selected from Agricultural Statistics Services databases and other sources and were enrolled between January 1994 and June 1996. Baseline data included detailed demographic, farm and farming, and behavioral information. From January 1994 to April 1998, subjects were contacted biannually to ascertain the occurrence of an agriculture-related injury. Injury rates were 2.9 times (95% confidence interval (CI): 2.0, 4.3) higher for African-American farm workers compared with Caucasian and African-American owners. Part-time farming (relative risk (RR) = 2.0, 95% CI: 1.3, 2.5), prior agricultural injury (RR = 1.5, 95% CI: 1.0, 2.1), and farm machinery in fair/poor condition (RR = 1.8, 95% CI: 1.2, 2.7) were also independently associated with injury rates. The results demonstrate the increased frequency of agricultural injury among farm workers and identify a number of possible ways of reducing them. PMID- 11032160 TI - Do common mental disorders increase cigarette smoking? Results from five waves of a population-based panel cohort study. AB - A prospective panel cohort design was used to investigate whether mental disorders common in the general population increase the likelihood of increased cigarette smoking at 12 months follow-up. By 1995, the last year for which data were available, a random sample of 12,057 persons aged 16-75 years residing in private households in Great Britain had been recruited. At each of five annual waves, the main exposure, past mental disorder, was derived from assessments of psychiatric morbidity as measured by the General Health Questionnaire-12. Increased cigarette smoking was derived from observations of number of cigarettes smoked and was defined by an increase of five or more per day relative to the previous calendar year. After logistic regression analysis, persons with a common mental disorder were about 30% more likely to have increased their cigarette smoking over the previous year (odds ratio = 1.29, 95% confidence interval: 1.16, 1.43). The estimated effect in the youngest (16-21 years) and oldest (51-75 years) age groups was higher than that in the middle (31-50 years) age group (odds ratios = 1.50, 1.57, and 1.12, respectively; test for interaction, chi2 = 6.8 (3 df), p = 0.078). These findings indirectly support the hypothesis that common mental disorders may have an enduring effect of increasing cigarette smoking a year later. PMID- 11032161 TI - Family history, maternal smoking, and clubfoot: an indication of a gene environment interaction. AB - Although epidemiologic studies of some birth defects have suggested a gene smoking interaction, the possibility of this interaction in clubfoot has not been examined. The authors analyzed risk factors among 346 infants with isolated clubfoot and 3,029 infants without defects from the Atlanta Birth Defects Case Control Study. All infants were born during 1968-1980, and mothers were interviewed in 1982-1983. The authors examined the family history-smoking interaction as an indication of a gene-environment interaction. They defined "smoking" as smoking any time during the first 3 months of pregnancy and "family history" as having a first-degree relative with clubfoot. Conditional logistic regression (matching variables: race, birth hospital, and birth period) was used to obtain effect estimates. The adjusted odds ratios were 1.34 (95% confidence interval (CI): 1.04, 1.72) for smoking only, 6.52 (95% CI: 2.95, 14.41) for family history only, and 20.30 (95% CI: 7.90, 52.17) for a joint exposure of smoking and family history. The effect estimate for the joint exposure was higher than would be expected under either an additive or a multiplicative model of interaction and showed a statistically significant departure from additivity. This study confirms the importance of familial factors and smoking in the etiology of clubfoot and identifies a potentially important interaction. PMID- 11032162 TI - Strong decline in herpes simplex virus antibodies over time among young homosexual men is associated with changing sexual behavior. AB - The objective of this study was to evaluate whether the change in sexual behavior among homosexual men observed after the start of the acquired immunodeficiency syndrome epidemic resulted in a change in herpes simplex virus (HSV) seroprevalence in this group over time. In a cross-sectional study, the prevalence of herpesvirus types 1 (HSV1) and 2 (HSV2) was determined at study entry in 1984-1985 and 1995-1997 among 532 young (aged < or = 30 years) homosexual men participating in the Amsterdam Cohort Studies on HIV/AIDS. Risk factors for the presence of HSV antibodies, including human immunodeficiency virus infection, were evaluated, and their influence on HSV prevalence over time was assessed. A strong decrease in HSV1 and HSV2 seroprevalence, from 80.6% to 59.0% and from 51.3% to 19.0%, respectively, was observed between the two time periods. This decrease was not markedly influenced by various demographic and socioeconomic factors. After data were controlled for several markers of sexual activity (such as number of sex partners, human immunodeficiency virus infection, and past episode(s) of gonorrhea), it appeared that the decline in HSV seroprevalence was explained by a concurrent decrease in the presence of these markers. The authors conclude that among young homosexual men in this study, the strong decrease in HSV seroprevalence was associated with a concurrent shift in sexual behavior. Furthermore, these data suggest an increasing sexual component in HSV1 transmission over time. PMID- 11032163 TI - Influenza vaccination and the risk of primary cardiac arrest. AB - Influenza epidemics are associated with an excess of mortality not only from respiratory diseases but also from other causes, and cardiovascular mortality increases abruptly during influenza epidemics, with little evidence of a lag period. In a population-based case-control study, the authors examined whether influenza vaccination was associated with a reduced risk of out-of-hospital primary cardiac arrest (PCA), a major contributor to cardiovascular mortality in the community. Cases of PCA (n = 342) without prior heart disease or life threatening comorbidity that occurred in King County, Washington, were identified from paramedic incident reports from October 1988 to July 1994. Demographically similar controls (n = 549) were identified from the community by using random digit dialing. Spouses of subjects were interviewed to assess treatment with influenza vaccine during the previous year and other risk factors. After adjustment for demographic, clinical, and behavioral risk factors, influenza vaccination was associated with a reduced risk of PCA (odds ratio = 0.51, 95 percent confidence interval: 0.33, 0.79). The authors suggest that while the association of influenza vaccination with a reduced risk of PCA is consistent with cohort studies of influenza vaccination and total mortality, further studies are needed to determine whether the observed association reflects protection or selection. PMID- 11032164 TI - Assessment of physical activity with a single global question in a large, multiethnic sample of midlife women. AB - This study compared responses from 13,621 African-American, Chinese, Hispanic, Japanese, and White women to a single, global physical activity question. Respondents aged 40-55 years were randomly selected from seven geographic locations in the United States for the 1996-1997 cross-sectional survey of the Study of Women's Health Across the Nation, a longitudinal, observational study of the menopause transition. Respondents rated their activity level as much less, less, the same as, more, or much more than other women their age. Physical activity rating varied minimally by race/ethnicity. The proportions of women who rated themselves much less active and much more active ranged from 3.1% for Whites to 4.8% for Japanese and from 13.6% for Japanese to 16.4% for African Americans, respectively. Multiple logistic regression models, stratified by race/ethnicity, showed independent associations between a low level of activity and higher body mass index, poor health, functional impairment, perceived stress, difficulty sleeping, and not being employed. A high level of activity was associated with excellent health, single marital status, higher education, lower body mass index, and older age. These findings suggest that a comparative rating of physical activity may rank women by activity level within a specific racial/ethnic group but may not capture differences across racial/ethnic groups. PMID- 11032165 TI - Re: "Problems due to small samples and sparse data in conditional logistic regression analysis". PMID- 11032166 TI - Re: "Asymptotic bias and efficiency in case-control studies of candidate genes and gene-environment interactions: basic family designs". PMID- 11032167 TI - Re: "Cigarette smoking and suicide: a prospective study of 300,000 male active duty army soldiers". PMID- 11032168 TI - Signal transduction mediated by Bid, a pro-death Bcl-2 family proteins, connects the death receptor and mitochondria apoptosis pathways. AB - Two major apoptosis pathways have been defined in mammalian cells, the Fas/TNF-R1 death receptor pathway and the mitochondria pathway. The Bcl-2 family proteins consist of both anti-apoptosis and pro-apoptosis members that regulate apoptosis, mainly by controlling the release of cytochrome c and other mitochondrial apoptotic events. However, death signals mediated by Fas/TNF-R1 receptors can usually activate caspases directly, bypassing the need for mitochondria and escaping the regulation by Bcl-2 family proteins. Bid is a novel pro-apoptosis Bcl-2 family protein that is activated by caspase 8 in response to Fas/TNF-R1 death receptor signals. Activated Bid is translocated to mitochondria and induces cytochrome c release, which in turn activates downstream caspases. Such a connection between the two apoptosis pathways could be important for induction of apoptosis in certain types of cells and responsible for the pathogenesis of a number of human diseases. PMID- 11032169 TI - The IAP family: endogenous caspase inhibitors with multiple biological activities. AB - IAPs (inhibitors of apoptosis) are a family of proteins containing one or more characteristic BIR domains. These proteins have multiple biological activities that include binding and inhibiting caspases, regulating cell cycle progression, and modulating receptor-mediated signal transduction. Our recent studies found the IAP family members XIAP and c-IAP1 are ubiquitinated and degraded in proteasomes in response to apoptotic stimuli in T cells, and their degradation appears to be important for T cells to commit to death. In addition to three BIR domains, each of these IAPs also contains a RING finger domain. We found this region confers ubiquitin protease ligase (E3) activity to IAPs, and is responsible for the auto-ubiquitination and degradation of IAPs after an apoptotic stimulus. Given the fact that IAPs can bind a variety of proteins, such as caspases and TRAFs, it will be of interest to characterize potential substrates of the E3 activity of IAPs and the effects of ubiquitination by IAPs on signal transduction, cell cycle, and apoptosis. PMID- 11032170 TI - Activation-induced cell death in B lymphocytes. AB - Upon encountering the antigen (Ag), the immune system can either develop a specific immune response or enter a specific state of unresponsiveness, tolerance. The response of B cells to their specific Ag can be activation and proliferation, leading to the immune response, or anergy and activation-induced cell death (AICD), leading to tolerance. AICD in B lymphocytes is a highly regulated event initiated by crosslinking of the B cell receptor (BCR). BCR engagement initiates several signaling events such as activation of PLCgamma, Ras, and PI3K, which generally speaking, lead to survival. However, in the absence of survival signals (CD40 or IL-4R engagement), BCR crosslinking can also promote apoptotic signal transduction pathways such as activation of effector caspases, expression of pro-apoptotic genes, and inhibition of pro-survival genes. The complex interplay between survival and death signals determines the B cell fate and, consequently, the immune response. PMID- 11032171 TI - Genetic regulation of programmed cell death in Drosophila. AB - Programmed cell death plays an important role in maintaining homeostasis during animal development, and has been conserved in animals as different as nematodes and humans. Recent studies of Drosophila have provided valuable information toward our understanding of genetic regulation of death. Different signals trigger the novel death regulators rpr, hid, and grim, that utilize the evolutionarily conserved iap and ark genes to modulate caspase function. Subsequent removal of dying cells also appears to be accomplished by conserved mechanisms. The similarity between Drosophila and human in cell death signaling pathways illustrate the promise of fruit flies as a model system to elucidate the mechanisms underlying regulation of programmed cell death. PMID- 11032172 TI - DNA fragmentation in apoptosis. AB - Cleavage of chromosomal DNA into oligonucleosomal size fragments is an integral part of apoptosis. Elegant biochemical work identified the DNA fragmentation factor (DFF) as a major apoptotic endonuclease for DNA fragmentation in vitro. Genetic studies in mice support the importance of DFF in DNA fragmentation and possibly in apoptosis in vivo. Recent work also suggests the existence of additional endonucleases for DNA degradation. Understanding the roles of individual endonucleases in apoptosis, and how they might coordinate to degrade DNA in different tissues during normal development and homeostasis, as well as in various diseased states, will be a major research focus in the near future. PMID- 11032173 TI - Effect of Bcl-2 and caspase-3 on calcium distribution in apoptosis of HL-60 cells. AB - Apoptosis manifests in two major execution programs downstream of the death signal: the caspase pathway and organelle dysfunction. An important antiapoptosis factor, Bcl-2 protein, contributes in caspase pathway of apoptosis. Calcium, an important intracellular signal element in cells, is also observed to have changes during apoptosis, which maybe affected by Bcl-2 protein. We have previously reported that in Harringtonine (HT) induced apoptosis of HL-60 cells, there's a change of intracellular calcium distribution, moving from cytoplast especially Golgi's apparatus to nucleus and accumulating there with the highest concentration. We report here that caspase-3 becomes activated in HT-induced apoptosis of HL-60 cells, which can be inhibited by overexpression of Bcl-2 protein. No sign of apoptosis or intracellular calcium movement from Golgi's apparatus to nucleus in HL-60 cells overexpressing Bcl-2 or treated with Ac-DEVD CHO, a specific inhibitor of caspase-3. The results indicate that activated caspase-3 can promote the movement of intracellular calcium from Golgi's apparatus to nucleus, and the process is inhibited by Ac-DEVD-CHO (inhibitor of caspase-3), and that Bcl-2 can inhibit the movement and accumulation of intracellular calcium in nucleus through its inhibition on caspase-3. Calcium relocalization in apoptosis seems to be irreversible, which is different from the intracellular calcium changes caused by growth factor. PMID- 11032174 TI - Nuclear apoptosis induced by isolated mitochondria. AB - We isolated and purified mitochondria from mouse livers and spinach leaves. When added into egg extracts of Xenopus laevis, they caused nuclei of mouse liver to undergo apoptotic changes. Chromatin condensation, margination and DNA ladder were observed. After incubating isolated mitochondria in some hypotonic solutions, and centrifuging these mixtures at high speed, we got mitochondrial supernatants. It was found that in the absence of cytosolic factor, the supernatant alone was able to induce apoptotic changes in nuclei. The effective components were partly of protein. DNA fragmentation was partly inhibited by caspase inhibitors AC-DEVD-CHO and AC-YVAD-CHO. Meanwhile, caspase inhibitors fully blocked chromatin condensation. Primary characterization of the nuclear endonuclease(s) induced by mitochondrial supernatants was also conducted. It was found that this endonuclease is different from endonuclease G, cytochrome c induced nuclease, or Ca2+-activated endonuclease. PMID- 11032175 TI - Expression of Bcl-2 inhibited Fas-mediated apoptosis in human hepatocellular carcinoma BEL-7404 cells. AB - Apoptosis plays an important role in embryonic development, tissue remodeling, immune regulation and tumor regression. Two groups of molecules (Bcl-2 family and "Death factor" family) are involved in regulating apoptosis. In order to know about the effect of Bcl-2 on apoptosis induced by Fas, a typical member of "Death factor" family, the transfection experiments with expression vectors pcDNA3-fl and pcDNA3-bcl-2 were performed in BEL-7404 cells, a human hepatocellular carcinoma cell line which expresses endogenous Fas, but not FasL and Bcl-2. The data showed that the expression of FasL in pcDNA3-fl transfected hepatoma cells obviously induced the apoptosis of the cells. However, the overexpression of Bcl 2 in pcDNA3-bcl-2 transfected 7404/b-16 cells counteracted pcDNA3-fl transient transfection mediated apoptosis. Further study by co-transfection experiments indicated that Bid but not Bax (both were pro-apoptotic proteins of Bcl-2 family) blocked the inhibitory effect of Bcl-2 on Fas-mediated apoptosis. These results suggested that Fas-mediated apoptosis in human hepatoma cells is possibly regulated by Bcl-2 family proteins via mitochondria pathway. PMID- 11032176 TI - Implications of peroxisome proliferator-activated receptors (PPARS) in development, cell life status and disease. AB - The past several years have seen an increasing interest in the peroxisome proliferator-activated receptors (PPARs). These transcriptional factors belong to the superfamily of the steroid/thyroid/retinoid receptors. They are activated by fatty acids or their metabolites as well as by different xenobiotic peroxisome proliferators. These receptors are expressed in both the embryo and the adult organism. They have been implicated in cell proliferation, differentiation and apoptosis. In this review, we will attempt to point out some of the more salient features of this expression pattern during development and the different steps of cell life. The current understanding of how PPARs are involved in some human diseases will also be described. PMID- 11032177 TI - Bone morphogenetic proteins (BMPs) induce epithelial differentiation of NT2D1 human embryonal carcinoma cells. AB - Human embryonal carcinoma (EC) cells represent the stem cells of testicular germ cell tumours (TGCTs) and are morphologically, antigenically and functionally related to the stem cells of early mammalian embryos. Despite the large capacity for differentiation displayed by TGCT stem cells, little is known of the factors controlling their developmental potency. We have analyzed the differentiation elicited in NT2D1 human embryonal carcinoma (EC) cells by Bone Morphogenetic Proteins (BMPs) and compared it with that elicited by retinoic acid (RA). We have found that while RA induced expression of neuronal, endodermal and epithelial markers in NT2D1 human EC cells, treatment with BMPs resulted in a predominantly epithelial phenotype. We also provide evidence to suggest that at least some of the effects elicited by RA in human EC cells might be mediated through RA-induced expression of BMP-7. Thus BMPs may play an important role in specifying the type of differentiation arising from human multipotent stem cells. The manipulation of BMP signalling in human embryonic multipotent stem cells may therefore prove a useful approach in attempts to generate specific differentiated cell types in vitro, and loss of the malignant and/or transformed phenotype. PMID- 11032178 TI - Effect of inducers and inhibitors on the expression of bcs genes involved in cypris larval attachment and metamorphosis of the barnacles Balanus amphitrite. AB - We examined the expression of six barnacle cypris larva-specific gene (bcs) cDNAs (bcs-1, -2, -3, -4,- 5, and -6), the bcs genes, by using Northern blot analysis under various conditions that induced or inhibited cypris larval attachment and metamorphosis. Inducers of larval attachment and metamorphosis, such as a neurotransmitter, tended to increase the expression of bcs mRNAs. All inhibitors of larval attachment and metamorphosis, such as G protein-coupled receptor agonists/antagonists, inhibitors of tyrosine kinase-linked receptors and inhibitors of their signal transduction, suppressed the expression of bcs-6 mRNA alone, but affected differentially other bcs genes. These results strongly suggest that the bcs-6 product plays a key role in triggering the attachment and metamorphosis of cypris larvae into juvenile barnacles. The roles of four late bcs genes (bcs-3,-4, -5 and -6) are discussed. PMID- 11032179 TI - Stage-dependent responses of the developing lung to retinoic acid signaling. AB - Morphological analysis of vitamin A-deficient rat fetuses and of retinoic acid receptor (RAR and RXR) mutant mice have demonstrated that retinoic acid (RA) is essential for lung development. To gainfurther insight into RA signaling pathways during primary lung budformation and lung branching, we have investigated the effects of RA and of a pan-RAR antagonist in cultures of whole embryos and lung explants. Treatment of E8.0 embryos with the pan-RAR antagonist inhibits the formation of the primitive respiratory system. On the other hand, treatment of E11.75 and E12.5 lung explants with RA inhibits branching morphogenesis, whereas treatment with the pan-RAR antagonist at the same developmental stages stimulates formation of distal buds. The inhibitory effect of RA on branching is strongly decreased in RARbeta null lungs, while enhancement of budding by the pan-RAR antagonist is not affected by an RARgamma null mutation. Additionally, cellular retinol binding protein one (CRBPI) null lungs are more sensitive than wild type lungs to the pan-RAR antagonist-induced stimulation of branching. These data indicate that retinoid signaling is indispensable for the formation of primary lung buds and the oesophagotracheal septum from the primitive foregut. They also suggest that at the pseudoglandular stage, RA signaling through RARbeta, but not RARgamma, inhibits distal bud formation thereby promoting the formation of conducting airways. Moreover, the level of CRBPI in the pseudoglandular lung appears to participate in the control of branching morphogenesis. PMID- 11032181 TI - Heat shock factor 2 is activated during mouse heart development. AB - Two members of the heat shock transcription factor family, HSF1 and HSF2, have been identified as activators of mammalian heat shock gene expression. HSF1 acts as a classical stress-responsive factor, whereas HSF2 might play a role in embryogenesis, since it is active during pre- and post-implantation periods up to 15.5 days of mouse embryonic development. In this study, we analyzed HSF1 and HSF2 expression and activation during mouse heart formation. Our results show an abundant expression of HSF1 throughout heart development. In contrast, expression of the alternatively spliced HSF2-alpha and HSF2-beta, and an additional higher molecular weight isoform is strongly upregulated in the developing mouse heart at E11.5-12.5, a stage after which tubular heart has looped and chambers formed, and the myocardial walls are maturating and the valves differentiating. At the same developmental stage, HSF2 DNA-binding activity is transiently induced, whereas the weak HSE-binding activity, which is detected throughout heart development, consists primarily of HSF1. Interestingly, heat shock gene expression shows no temporal or spatial correlation with HSF2 expression and activation. Taken together, our results indicate that HSF2 activation is associated with specific stages of heart formation but is not involved in the regulation of inducible heat shock gene expression. PMID- 11032180 TI - Retinoid signalling acts during the gastrula stages to promote primary neurogenesis. AB - Retinoid signalling has been manipulated at different developmental stages to identify a critical period in the gastrula embryo for retinoid-dependent primary neurone formation. The expression of retinoid receptor RARalpha2 in the posterior neuroectoderm of the gastrula embryo is therefore consistent with a role in primary neurogenesis. In addition we show that the expression of neurogenin-1 and XDelta-1, two genes that contribute to the determination of primary neurone cell fate in the gastrula embryo, respond to retinoid signalling. These results indicate that retinoid signalling is required for an early step in the process of primary neurogenesis. When retinoid signalling is increased, the number of primary neurones increases, but the phenotype is not the same as the neurogenic phenotype that follows the overexpression of a dominant negative form of XDelta 1. Whereas increased retinoid signalling expands the width of primary neurone stripes, dominant negative XDelta-1 increases the density of primary neurones within the stripes. When retinoid signalling is increased and the primary neurone stripes expand, the expression domain of a floorplate marker contracts. Conversely, when retinoid signalling is inhibited, the expression patterns of floorplate markers widen. These results indicate that retinoid signalling acts at an early stage in primary neural development when the fates of different regions of the neuroectoderm are being determined. PMID- 11032182 TI - Expression of the cardiac actin gene in axolotl embryos. AB - Axolotis are an important model system for studying heart development. Patterning of the somitic mesoderm occurs in axolotis in a manner that is much more similar to the pattern observed in higher vertebrates than in Xenopus. For these reasons we cloned the axolotl cardiac actin gene, since this gene is expressed during the development of both somitic and cardiac muscle in other vertebrates. In this paper we characterize its expression. Expression of cardiac actin RNA is switched on during gastrula stages and appears in the somitic mesoderm when it is formed; expression is later activated in the embryonic heart. In adults the gene is expressed only in the heart. The results demonstrate that the clone encoding cardiac actin provides a useful marker for studying development of both skeletal and cardiac muscle development in axolotls. PMID- 11032183 TI - Induction of segmentation in polyps of Aurelia aurita (Scyphozoa, Cnidaria) into medusae and formation of mirror-image medusa anlagen. AB - Polyps of Aurelia aurita can transform into several medusae (jellyfish) in a process of sequential subdivision. During this transformation, two processes take place which are well known to play a key role in the formation of various higher metazoa: segmentation and metamorphosis. In order to compare these processes in bilaterians and cnidarians we studied the control and the kinetics of these processes in Aurelia aurita. Segmentation and metamorphosis visibly start at the polyp's head and proceed down the body column but do not reach the basal disc. The small piece of polyp which remains will develop into a new polyp. The commitment to the medusa stage moves down the body column and precedes the visible onset of segmentation by about one day. Segmentation and metamorphosis can start at the cut surface of transversely cut body columns, leading to a mirror-image pattern of sequentially developing medusae. PMID- 11032184 TI - A cell type-specific effect of calcium on pattern formation and differentiation in dictyostelium discoideum. AB - Spatial gradients of sequestered and free cellular calcium (Ca2+) exist in the slug of Dictyostelium discoideum (Maeda and Maeda, 1973; Tirlapur et al., 1991; Azhar et al., 1995; Cubitt et al., 1995). When we vary intracellular Ca2+ with the help of calcium buffers and the ionophore Br-A23187, there are striking effects on slug morphology, patterning and cell differentiation. In the presence of a calcium ionophore, high external Ca2+ levels lead to an increase of intracellular sequestered and free Ca2+, the formation of long slugs, a decrease in the fraction of genetically defined prespore cells and 'stalky' fruiting bodies. Conversely, a lowering of external Ca2+ levels results in a decrease of intracellular Ca2+, the formation of short slugs, an increase in the prespore fraction and 'spory' fruiting bodies. We infer that Ca2+ plays a significant morphogenetic role in D. discoideum development, by selectively promoting the prestalk pathway relative to the prespore pathway. PMID- 11032185 TI - Comparative approach to developmental analysis: the case of the dalyellid flatworm, Gieysztoria superba. AB - Dalyellida represents a taxon of small rhabdocoel flatworms that occur in freshwater habitats all over the world. Combining histology and electron microscopy we have analyzed the embryonic development of a new dalyellid species, Gieysztoria superba, in order to obtain more comparative data pertaining to morphogenesis and organogenesis in flatworms. We have used a morphological staging system that we recently introduced for another rhabdocoel, Mesostoma lingua (Younossi-Hartenstein et al., 2000). Our data show that in many fundamental respects, such as the irregular cleavage, mesenchymal embryonic primordium, and lack of gastrulation movements, Gieysztoria is highly similar to Mesostoma. During cleavage (stages 1 and 2) the embryo is located in the center of the egg where it is surrounded by a layer of yolk cells. Cleavage leads up to a solid, disc shaped cell cluster. During stage 3, the embryo migrates to the ventral side of the egg and acquires bilateral symmetry. Stages 4/5 sees the emergence of the first organ primordia, the brain, epidermis and pharynx. A peculiar invagination of the epidermal layer pushes the embryo back into the center of the yolk ("embryonic invagination"). Organogenesis takes place during stages 5 and 6 while the embryo is invaginated. A junctional complex, consisting initially of small septate junctions, followed later by a more apically located zonula adherens, is formed in all epithelial tissues, including epidermis, protonephridia, and pharynx. During late stages (6-8), Gieysztoria embryos evert back to the surface where the epidermal primordium expands and grows around the yolk to close dorsally. During this phase of development cytodifferentiation of the different organ systems takes place. Stage 7 is characterized by the appearance of eye pigmentation, brain condensation and spindle shaped myocytes. Stage 8 describes the fully dorsally closed and differentiated embryo. In comparison to other rhabdocoels, including Mesostoma, Gieysztoria exhibits a precocious differentiation of an intestinal epithelium and male genital apparatus. In Mesostoma, these structures are formed post hatching. PMID- 11032186 TI - Overexpression of S-adenosylmethionine decarboxylase (SAMDC) activates the maternal program of apoptosis shortly after MBT in Xenopus embryos. AB - Overexpression of S-adenosylmethionine decarboxylase (SAMDC) mRNA in 1- and 2 cell stage Xenopus embryos induces cell autonomous dissociation at the late blastula stage and developmental arrest at the early gastrula stage. The induction of cell dissociation took place "punctually" at the late blastula stage in the SAMDC-overexpressing cells, irrespective of the stage of the microinjection of SAMDC mRNA. When we examined the cells undergoing the dissociation, we found that they were TUNEL-positive and contained fragmented nuclei with condensed chromatin and fragmented DNA. Furthermore, by injecting Xenopus Bcl-2 mRNA together with SAMDC mRNA, we showed that SAMDC-overexpressing embryos are rescued completely by Bcl-2 and becometadpoles. These results indicatethat cell dissociation induced by SAMDC overexpression is due to apoptotic cell death. Since the level of S-adenosylmethionine (SAM) is greatly reduced in SAMDC-overexpressing embryos and this induces inhibition of protein synthesis accompanied by the inhibition of DNA and RNA syntheses, we conclude that deficiency in SAM induced by SAMDC overexpression activates the maternal program of apoptosis in Xenopus embryos at the late blastula stage, but not before. We propose that this mechanism serves as a surveillance mechanism to check and eliminate cells physiologically damaged during the cleavage stage. PMID- 11032187 TI - Vaccines across the life span. Teaching Immunization for Medical Education (TIME) Project. PMID- 11032188 TI - Haemophilus influenzae type B disease, vaccines, and care of exposed individuals. AB - Before effective vaccines became available, approximately 1 in every 200 children aged younger than 5 years had invasive Haemophilus influenzae type b (Hib) disease. Hib was the most common cause of bacterial meningitis and other invasive bacterial diseases in this age group. Rapid diagnosis and treatment are essential for Hib meningitis, because the mortality rate is 2% to 5%, even with antibiotic treatment--usually a third-generation cephalosporin, such as cefotaxime or ceftriaxone. Because of the use of Hib vaccines, the incidence of invasive H. influenzae disease in children younger than 5 years old declined by 97% between 1987 and 1997. Recent data indicate that the conjugate Hib vaccines given in infancy can be used interchangeably. PMID- 11032189 TI - Prevention of influenza by expanded ages for routine vaccination. AB - Influenza viruses are highly contagious and are transmitted from person to person, usually by the airborne route. Persons in semiclosed or crowded environments, such as students and residents of nursing homes, are at high risk for exposure. Fatality rates are highest in persons who have chronic medical conditions such as chronic obstructive lung disease and diabetes mellitus, particularly if they are elderly. When there is a good match between the vaccine and the circulating viruses, influenza vaccine has been shown to prevent illness in approximately 70% to 90% of healthy persons younger than 65 years. Despite the availability of an effective vaccine, it is underused. The Advisory Committee on Immunization Practices (ACIP) and the American Academy of Family Physicians (AAFP) now recommend that all persons aged 50 years and older receive annual influenza vaccination because of the suffering from influenza and the cost effectiveness of vaccination. Reasons for lowering the recommended age for routine vaccination from 65 years to 50 years include reductions in office visits, hospitalizations, time taken off work, and costs. Persons younger than 50 years who have medical conditions that place them at risk for complications should also be vaccinated. If a vaccine shortage occurs, which may happen in the Fall 2000, then priority would be given to the elderly and those with high-risk conditions. PMID- 11032190 TI - Child vaccination, part 1: routine vaccines. AB - Despite the success of the national childhood vaccination program in the United States in decreasing mortality due to vaccine-preventable diseases, vaccination rates remain suboptimal. Contributing factors include the failure to appreciate the hazards of vaccine-preventable diseases, concerns about adverse reactions associated with vaccine administration, and missed opportunities to administer vaccines. The 2 major types of indications for vaccinating children are age and presence of a medical condition that increases the risk of a vaccine-preventable disease. Hepatitis B virus (HBV) infection becomes chronic in 90% of those infected as infants, and 25% of those so infected will die of related chronic liver disease as adults. Routine infant vaccination against hepatitis B has been recommended since 1991. Approximately 69% of infants who develop pertussis require hospitalization. Acellular pertussis vaccines have been licensed for use in infancy. Starting in 2000, the all-inactivated poliovirus vaccine (IPV) schedule is recommended. IPV should eliminate vaccine-associated paralytic poliomyelitis. Pneumococcal conjugate vaccine was licensed in 2000 for routine use on a schedule of 2, 4, 6, and 12 to 15 months. The first dose of measles mumps-rubella vaccine is now recommended at age 12 to 15 months, simultaneous with varicella vaccine administration. PMID- 11032191 TI - Child vaccination, part 2: childhood vaccination procedures. AB - In 1996, the Advisory Committee on Immunization Practices (ACIP), the American Academy of Family Physicians (AAFP), the American Academy of Pediatrics (AAP), and the American Medical Association recommended a well-child office visit at age 11 to 12 years to check vaccination status. Vaccination status should be assessed for varicella, hepatitis B, the second dose of measles-mumps-rubella (MMR) vaccine, and tetanus-diptheria (Td) toxoid if not given in the past 5 years. Adolescent patients should be screened for high-risk conditions indicating the need for influenza, pneumococcal, or hepatitis A vaccines. The Accelerated Immunization Schedule and Minimal Interval Table should be consulted for children who are behind schedule. PMID- 11032192 TI - Adult vaccination, part 1: vaccines indicated by age. Teaching Immunization for Medical Education (TIME) Project. AB - The morbidity and mortality of vaccine-preventable diseases among adults are high, particularly among populations at high risk because of underlying medical conditions. Influenza vaccination is recommended annually, optimally during campaigns held between October and mid-November, for all persons 50 years and older and for younger persons with high-risk conditions. Because of production delays, influenza vaccination campaigns are delayed until November of this year. Pneumococcal polysaccharide vaccination is recommended for healthy persons 65 years and older and younger persons with high-risk conditions. A 3-dose series of adult tetanus and diphtheria toxoids (Td) is recommended for those who have not had a primary series or whose vaccination history is uncertain. Adults who have completed the primary vaccination series should receive a booster dose of Td vaccine every 10 years. Specific strategies for improving the rate of these vaccinations have been developed for medical offices and clinics, hospitals, and other health care institutions, and other settings where there is high risk of vaccine-preventable disease. PMID- 11032193 TI - Adult vaccination, part 2: vaccines for persons at high risk. Teaching Immunization for Medical Education (TIME) Project. AB - The morbidity and mortality from vaccine-preventable diseases are high among adults with underlying medical conditions. Influenza vaccination is recommended annually, optimally between October and mid-November, for all persons 50 years of age and older and those with cardiac disease with potential for altered hemodynamics, diabetes mellitus, immunocompromising conditions, pulmonary disease, or renal disease. This season, because of production delays, influenza vaccination campaigns are planned for November. Pneumococcal polysaccharide vaccination is recommended for all persons 65 years and older and for those with alcoholism, asplenia, cardiac disease, cirrhosis, diabetes mellitus, immunocompromising conditions, pulmonary disease, or chronic renal disease. Indications for hepatitis B vaccination include chronic renal disease and hemodialysis, as well as employment in health care or employment as a mortician or public safety officer. It is also recommended for homosexual men, those who have multiple sex partners or a sexually transmitted disease, and injection drug users. PMID- 11032194 TI - How should we determine the best treatment for patients with asymptomatic carotid stenosis? PMID- 11032195 TI - How safe and effective are nonsteroidal anti-inflammatory drugs (NSAIDs) in the treatment of acute or chronic nonspecific low back pain (LBP)? PMID- 11032196 TI - Is it always necessary to suture all lacerations after a vaginal delivery? PMID- 11032198 TI - How accurate are the history and physical examination in diagnosing carpal tunnel syndrome (CTS)? PMID- 11032197 TI - Are high-dose inhaled steroids effective for chronic obstructive pulmonary disease (COPD)? PMID- 11032199 TI - Does delayed pushing reduce difficult deliveries for nulliparous women with epidural analgesia? PMID- 11032200 TI - Is oral zolmitriptan efficacious in the acute treatment of cluster headache? PMID- 11032201 TI - Training in back care to improve outcome and patient satisfaction. Teaching old docs new tricks. AB - BACKGROUND: We examined clinical outcomes and patient perceptions of back care given by physicians before and after an intensive course of training in back care and limited manual therapy techniques. METHODS: From a prospective observational cohort study of low back pain involving 208 physicians (115 primary care) and their patients and a subsequent clinical trial of treatment of low back pain given by 31 physicians specially trained in manual therapy and enhanced back care, outcome data from the patients of 13 physicians participating in both studies were compared. In the observational study, the 13 physicians cared for 120 patients. In the manual therapy trial (191 patients) a control group of 94 patients received enhanced back care and an intervention group of 97 patients received enhanced back care plus manual therapy. Pearson's chi-square comparisons and linear and Cox proportional hazard modeling were used to examine effects of variables and recovery time. RESULTS: Characteristics of the 13 physicians' patients in the cohort group and the manual therapy trial showed some differences in income, workers' compensation, previous employment, and baseline dysfunction. Both control and intervention patients in the manual therapy trial showed more rapid improvement in functional status over time and greater satisfaction with their care than those in the previous cohort study. However, there was no difference between the studies in patient-reported time to return to performing usual daily activities. CONCLUSIONS: A structured clinical approach to low back care may bring modestly improved clinical outcomes and patient satisfaction. PMID- 11032202 TI - Understanding and treating low back pain in family practice. PMID- 11032203 TI - The impact of patient-centered care on outcomes. AB - BACKGROUND: We designed this observational cohort study to assess the association between patient-centered communication in primary care visits and subsequent health and medical care utilization. METHODS: We selected 39 family physicians at random, and 315 of their patients participated. Office visits were audiotaped and scored for patient-centered communication. In addition, patients were asked for their perceptions of the patient-centeredness of the visit. The outcomes were: (1) patients' health, assessed by a visual analogue scale on symptom discomfort and concern; (2) self-report of health, using the Medical Outcomes Study Short Form-36; and (3) medical care utilization variables of diagnostic tests, referrals, and visits to the family physician, assessed by chart review. The 2 measures of patient-centeredness were correlated with the outcomes of visits, adjusting for the clustering of patients by physician and controlling for confounding variables. RESULTS: Patient-centered communication was correlated with the patients' perceptions of finding common ground. In addition, positive perceptions (both the total score and the subscore on finding common ground) were associated with better recovery from their discomfort and concern, better emotional health 2 months later, and fewer diagnostic tests and referrals. CONCLUSIONS: Patient-centered communication influences patients' health through perceptions that their visit was patient centered, and especially through perceptions that common ground was achieved with the physician. Patient-centered practice improved health status and increased the efficiency of care by reducing diagnostic tests and referrals. PMID- 11032204 TI - The science of patient-centered care. PMID- 11032206 TI - Physicians, pharmaceutical representatives, and patients: who really benefits? PMID- 11032205 TI - The value of pharmaceutical representative visits and medication samples in community-based family practices. AB - BACKGROUND: Interactions between the pharmaceutical industry and physicians have been discussed in numerous publications; however, most articles are limited to surveys and self-report data and often focus on academic or training contexts. We describe the role of pharmaceutical representatives and the use of samples in community-based family practices, using data obtained by directly observing clinical encounters. METHODS: We collected detailed descriptive field notes of the direct observations of 53 primary care clinicians and 1588 patient encounters in 18 purposefully selected Nebraska family practices. We used a comparative case study design, that used depth interviews of clinicians and office staff, and included details of the interactions with pharmaceutical representatives and the use of samples in clinical encounters. RESULTS: Individual providers and practices displayed noticeable variation in their approaches to drug representatives and samples. We found formal strategies and policies in a minority of practices. Generally there was little structure in the organization and distribution of sample medications at the office level, and detailed patient education regarding these drugs was rarely observed in patient encounters. Nevertheless, samples were used in almost 20% of observed encounters, at times as starter dosages, but often as complete courses of treatment. The benefits derived from contact with the pharmaceutical industry varied substantially, but most often included free medication samples, meals, and patient education materials. CONCLUSIONS: Clinicians have a complex symbiosis with the pharmaceutical industry and need to critically evaluate their handling of samples and their contact with pharmaceutical representatives to optimize this relationship and ensure quality patient care. Clinics with specific policies for interactions with drug companies appear to derive more satisfaction from their encounters. PMID- 11032207 TI - Lifetime patterns of contraception and their relationship to unintended pregnancies. AB - BACKGROUND: For the past 30 years many effective methods of contraception have been available, yet unintended pregnancy rates still range from 30% to 50% in many populations. We examined patterns of women's contraceptive use throughout their lives and relate them to unintended pregnancy. METHODS: A total of 396 women aged 18 to 50 years chosen by convenience sampling from a family practice residency office were interviewed in a cross-sectional study about their history of using and changing contraception, and whether they believed they became pregnant while using a method of contraception. We analyzed the data for correlations and significance using chi-square and Student t tests. RESULTS: Most women had used both condoms and oral contraceptive pills, and tried an average of 3.54 methods during a lifetime. Two patterns of women's use of contraception emerged that describe 75% of the women. One third of the women--those who indicated a pattern of following their first method with a less effective method- are significantly more likely to have an unintended pregnancy while using contraception (odds ratio=1.4). The other group (50% of the entire sample) used increasingly effective methods and were less likely to have an unintended pregnancy. CONCLUSIONS: Pregnancy is an inherent natural consequence of sexual intercourse, even when using very effective contraceptive methods. By asking a few questions about a woman's history of contraceptive use, physicians may be able to determine those who are more likely to be at risk for an unintended pregnancy. PMID- 11032208 TI - Using ferritin levels to determine iron-deficiency anemia in pregnancy. AB - BACKGROUND: Standard obstetrical practice has included iron therapy for patients with anemia without requiring the determination of iron deficiency. However, the proportion of pregnant women with anemia who have such a deficiency may be relatively modest. We instituted a practice protocol using serum ferritin levels to determine the proportion of women undergoing prenatal care who had both anemia and iron deficiency. METHODS: We evaluated consecutive women entering prenatal care. Those with anemia (hemoglobin level <11 mg/dL) underwent testing for serum ferritin level and other hematologic variables. RESULTS: A total of 182 patients entered prenatal care. Hemoglobin data were available for 173 (95%). Thirty-eight (22%) had anemia (hemoglobin level <11 mg/dL); 1 of those women was excluded from the study. Using a ferritin level of 12 mg per dL as the cutoff for iron deficiency, 54% (20) of the 37 remaining patients with anemia had an iron deficiency, and 46% (17) had anemia not related to such a deficiency. Use of hematologic indices provided on complete blood count were not useful in predicting iron deficiency based on serum ferritin levels. CONCLUSIONS: In our population of prenatal patients with anemia, only approximately half had an iron deficiency. Diagnostic and therapeutic approaches to screening for anemia in pregnancy should be reconsidered and further evaluated. PMID- 11032209 TI - The relationship between volume status, hydration, and radiographic findings in the diagnosis of community-acquired pneumonia. AB - BACKGROUND: Many clinicians believe the radiographic expression of community acquired pneumonia (CAP) is affected by the fluid volume status of the patient. However, there are very few data to support or refute this concept. With this study we began to examine the relationship between admission fluid volume status and the radiographic expression of CAP. METHODS: Using a retrospective chart review, we examined 376 consecutive inpatient encounters with the diagnosis of pneumonia at discharge from a community teaching hospital. Patients were evaluated by age, sex, admission serum sodium, blood urea nitrogen (BUN) level, creatinine, and fluid administered in the first 48 hours of treatment. We classified these patients as either showing radiographic progression (P) or no radiographic progression (NP) by comparison of admission and follow-up radiographs. RESULTS: A total of 125 patient encounters satisfied inclusion criteria for the study. Using the Student t test we noted a statistically significant difference between the P and NP groups for BUN level (P=.02), volume of fluid administered during the first 48 hours (P=.04), and marginally for age (P=.05). The P group had higher BUN levels (mean=34 vs 24), more 48-hour fluid intake (mean=5824 mL vs 4764 mL), and younger age (mean=59 years vs 66 years) than the NP group. Logistic regression poorly predicted which patients would have worsening infiltrate on the second radiograph. CONCLUSIONS: Elevated admission BUN level and higher fluid volume administered in the first 48 hours of admission were associated with worsening radiographic findings of pneumonia after hydration. Prospective studies are needed for confirmation of our results. PMID- 11032210 TI - Bisphosphonates in the prevention and treatment of glucocorticoid-induced osteoporosis. AB - OBJECTIVE: To summarize the literature concerning the use of bisphosphonates in the prevention and treatment of corticosteroid-induced osteoporosis and make recommendations concerning the proper use of these agents. SEARCH STRATEGIES: We conducted a literature search to identify studies in the English language concerning the use of bisphosphonates in the prevention or treatment of corticosteroid-induced osteoporosis using the MEDLINE, CURRENT CONTENTS, and HEALTHSTAR electronic databases, bibliographies of selected citations, and recent meeting abstracts. SELECTION CRITERIA: We included randomized controlled trials evaluating the use of oral bisphosphonates in adults by central dual X-ray absorptiometry. DATA COLLECTION AND ANALYSIS: We assessed the methodologic quality of the trials using the Jadad criteria. Data were collected concerning bone mineral density (BMD) changes in multiple areas, fracture rates, safety, and tolerability. MAIN RESULTS: Bisphosphonates generally increased BMD at the lumbar spine. Data were less clear concerning changes in the femoral area. Little information exists about the ability of bisphosphonates to reduce fracture risk in patients with corticosteroid-induced osteoporosis. Postmenopausal women seemed to receive the most benefit. CONCLUSIONS: Bisphosphonates significantly increased BMD in patients at risk for corticosteroid-induced bone loss. However, there is a sparsity of data concerning the ability of these agents to affect the clinically important outcome of fracture rate reduction, especially among premenopausal women in whom fractures are rare within the first year or 2 of exposure to corticosteroids. Long-term studies powered to detect fracture risk reduction are needed as well as comparative trials with bisphosphonates and other agents. PMID- 11032211 TI - Is exposure to HMG-CoA reductase inhibitors (statins), fibrates, or other lipid lowering drugs associated with reduced risk of bone fracture in older patients? PMID- 11032212 TI - The effectiveness of screening mammography. PMID- 11032213 TI - Hydrocele puncture during vasectomy. PMID- 11032214 TI - A rock and a hard place. PMID- 11032215 TI - It really bothers me. PMID- 11032216 TI - Influence of resistance training volume and periodization on physiological and performance adaptations in collegiate women tennis players. AB - Few data exist on the long-term adaptations to heavy resistance training in women. The purpose of this investigation was to examine the effect of volume of resistance exercise on the development of physical performance abilities in competitive, collegiate women tennis players. Twenty-four tennis players were matched for tennis ability and randomly placed into one of three groups: a no resistance exercise control group, a periodized multiple-set resistance training group, or a single-set circuit resistance training group. No significant changes in body mass were observed in any of the groups throughout the entire training period. However, significant increases in fat-free mass and decreases in percent body fat were observed in the periodized training group after 4, 6, and 9 months of training. A significant increase in power output was observed after 9 months of training in the periodized training group only. One-repetition maximum strength for the bench press, free-weight shoulder press, and leg press increased significantly after 4, 6, and 9 months of training in the periodized training group, whereas the single-set circuit group increased only after 4 months of training. Significant increases in serve velocity were observed after 4 and 9 months of training in the periodized training group, whereas no significant changes were observed in the single-set circuit group. These data demonstrate that sport-specific resistance training using a periodized multiple-set training method is superior to low-volume single-set resistance exercise protocols in the development of physical abilities in competitive, collegiate women tennis players. PMID- 11032217 TI - Long-term prognosis of patients with achilles tendinopathy. An observational 8 year follow-up study. AB - To determine the long-term outcome of patients treated nonoperatively for acute or subchronic (duration of the symptoms before initiation of the treatment less than 6 months) Achilles tendinopathy, we performed a follow-up analysis on 83 of 107 patients an average 8 +/- 2 (SD) years after the initial contact. The analysis included a questionnaire, clinical examination, performance tests, muscle strength measurement, and ultrasonographic examination. Twenty-four of the 83 patients (29%) had to be operated on during the follow-up period. Seventy patients (84%) had full recovery of their activity level, and at 8 years' follow up 78 patients (94%) were asymptomatic or had only mild pain with strenuous exercise. However, a clear side-to-side difference between the involved and the uninvolved sides was observed on the performance test, clinical examination, and ultrasonography. Also, 34 patients (41%) started to suffer from overuse symptoms in the initially uninvolved Achilles tendon. The results of our 8-year follow-up showed that the long-term prognosis of patients with acute-to-subchronic Achilles tendinopathy is favorable as determined by subjective and functional assessments. In the clinical and ultrasonographic examinations, mild-to-moderate changes were observed rather frequently in both the involved and initially uninvolved Achilles tendons, but the occurrence of these changes was not clearly related to the patients' symptoms. PMID- 11032218 TI - Epidemiology of concussion in collegiate and high school football players. AB - Despite evolutionary changes in protective equipment, head injury remains common in football. We investigated concussion in football and associated epidemiologic issues such as 1) incidence of injury, 2) common signs and symptoms, and 3) patterns in making return-to-play decisions. We received 242 of 392 surveys (62%) that were sent to high school and collegiate certified athletic trainers at the beginning of three football seasons. Of the 17,549 football players represented, 888 (5.1%) sustained at least one concussion, and 131 (14.7% of the 888) sustained a second injury during the same season. The greatest incidence of concussion was found at the high school (5.6%) and collegiate division III (5.5%) levels, suggesting that there is an association between level of play and the proportion of players injured. Players who sustained one concussion in a season were three times more likely to sustain a second concussion in the same season compared with uninjured players. Contact with artificial turf appears to be associated with a more serious concussion than contact with natural grass. Only 8.9% of all injuries involved loss of consciousness, while 86% involved a headache. Overall, 30.8% of all players sustaining a concussion returned to participation on the same day of injury. PMID- 11032219 TI - Primary anterior cruciate ligament reconstruction using the contralateral autogenous patellar tendon. AB - We studied patients who underwent primary anterior cruciate ligament reconstruction using either the contralateral (N = 434) or ipsilateral (N = 228) autogenous patellar tendon graft to determine the difference between groups for the return of range of motion, quadriceps muscle strength, and return to sports. The contralateral group had statistically significantly more flexion than the ipsilateral group at 1 week and 2 weeks postoperatively. The contralateral group had statistically significantly greater quadriceps muscle strength in the reconstructed knee at 1, 2, and 4 months postoperatively and in the donor knee at 1 and 2 months postoperatively. Mean KT-1000 arthrometer results were 1.9 +/- 1.3 mm for the contralateral group and 2.2 +/- 1.1 mm for the ipsilateral group. The mean time to return to sports at full capability in a competitive subgroup was 4.1 months for contralateral patients and 5.5 months for ipsilateral patients. Overall, 49% of patients in the contralateral group and 12% of patients in the ipsilateral group returned to their preinjury levels of activity by 4 months postoperatively. Our results indicate that the contralateral patellar tendon can be used to restore range of motion and strength sooner than an ipsilateral patellar tendon graft. Patients can also have a faster return to full capability in sports without compromising ultimate stability. PMID- 11032220 TI - Avoidance of soccer injuries with preseason conditioning. AB - The effect of a preseason conditioning program was studied to evaluate its influence on the occurrence and severity of soccer injuries. Three hundred female soccer players (ages 14 to 18 years) were studied over a 1-year period. Forty-two of these players participated in a 7-week training program before the start of the season. The type, mechanism, and severity of the injury, when the injury occurred, the number of games or practices missed, and type of shoe worn were recorded. All injuries occurred in the lower extremities, with 61.2% occurring at the knee and ankle. Student's t-test evaluations revealed that the trained group experienced a significantly lower incidence of injury than the untrained group (P = 0.0085). Although not statistically significant, the trained group also had a lower percentage (2.4%) of anterior cruciate ligament injuries compared with the untrained group (3.1%). These results suggest that this type of conditioning has a significant influence on lowering the incidence of injury in female adolescent soccer players. PMID- 11032221 TI - Clinical outcome of isolated subcortical trabecular fractures (bone bruise) detected on magnetic resonance imaging in knees. AB - Isolated subcortical trabecular bone injury (bone bruise) has rarely been described. Our purpose is to report a series of patients who had a history of traumatic injury, knee effusion, normal radiographs, and initial equivocal physical examination for ligament and meniscal integrity, and who were found to have isolated injury of the trabecular bone on magnetic resonance imaging. We evaluated demographic data, physical examination findings, radiographs, magnetic resonance imaging, and clinical outcome for 23 patients. Follow-up data included time to return to preinjury activity level, International Knee Documentation Committee activity level rating before and after injury, and postinjury Lysholm scores. All magnetic resonance imaging scans were negative for associated grade III meniscal lesions and ligament injury. Time to return to preinjury activity level was under 7 months in 96% of the patients. Postinjury International Knee Documentation Committee rating was unchanged in 91% of patients. Postinjury Lysholm score was 90 or more in 91% of patients. We propose that the recognition of these injuries is important because magnetic resonance imaging can distinguish them from meniscal or ligament injury requiring surgical intervention (arthroscopy). If detected on magnetic resonance imaging as an isolated injury, surgical arthroscopy is unnecessary since these patients can be expected to recover well in the short term with restricted weightbearing and initial activity modification. PMID- 11032222 TI - Quantification of posterior capsule tightness and motion loss in patients with shoulder impingement. AB - The relationship between posterior capsule tightness and dysfunction has long been recognized clinically but has not been biometrically quantified. The purpose of this study was to quantify changes in range of motion and posterior capsule tightness in patients with dominant or nondominant shoulder impingement. Measurements of posterior capsule tightness and external and internal rotation range of motion were made in 31 patients with shoulder impingement and in 33 controls without shoulder abnormality. Patients with impingement in the nondominant arm had increased posterior capsule tightness and decreased internal and external rotation range of motion compared with controls. Patients with impingement in their dominant arm had increased posterior capsule tightness and reduced internal rotation range of motion but no significant loss of external rotation range of motion compared with controls. Posterior capsule tightness in impingement patients showed a significant correlation with loss of internal rotation range of motion. Patients with shoulder impingement in their nondominant arm had a more global loss of range of motion compared with patients having impingement in their dominant arm. We believe we have described a valid clinical measurement for identifying posterior capsule tightness in patients with shoulder impingement. PMID- 11032223 TI - The effect of circumferential taping on flexor tendon pulley failure in rock climbers. AB - The purpose of this study was to determine whether circumferential taping of the base of the finger increases the A2 pulley's load to failure in a model simulating a rock climber's grip. Nine pairs of fresh-frozen cadaveric hands, 20 to 47 years of age, were rigidly mounted in a specialized jig that maintained the finger in the climber's "crimp" position. Two of the four fingers of each hand were reinforced over the A2 pulley with three wraps of cloth adhesive tape. The flexor digitorum profundus and superficialis tendons were distracted until pulley or tendon failure. Overall, A2 pulley strength was greater in male specimens than in female specimens, and the A2 pulley of the small finger was the weakest tested. The A2 pulley failed simultaneously with the A3 and A4 pulleys in 55% of the tests. In the remaining trials, a single pulley failed initially followed by the remainder of the sheath. Of the 72 fingers studied, complete data were available for comparison of 22 pairs of fingers. No statistically significant difference in load to A2 pulley failure was noted between the taped and untaped finger pairs. Based on our findings we do not support taping the base of the fingers as a prophylactic measure against flexor tendon sheath injury in the climbing athlete. PMID- 11032224 TI - Chronic calf pain in athletes due to sural nerve entrapment. A report of 18 cases. AB - We retrospectively analyzed the charts of 13 athletes (18 limbs) who had sural nerve entrapment localized in the passage of the nerve through the superficial sural aponeurosis. There were 11 men and 2 women (average age, 43 years; range, 31 to 59). All patients reported chronic calf pain that was exacerbated during physical exertion. Delay to diagnosis averaged 9 months (range, 5 to 24). Tenderness in the calf was identified along the course of the sural nerve in all cases. In 10 patients (15 limbs) electrodiagnostic testing before surgery was positive. After failure of nonoperative treatment, surgery was conducted under local anesthesia. Neurolysis was performed by incising the superficial sural aponeurosis and the fibrous band in it through which the nerve passes. The results of the operation were evaluated in terms of residual symptoms, ability to return to the former sport, and degree of patient satisfaction. A final follow-up examination was performed an average of 14 months (range, 6 to 30) after the operation. The final result was excellent in 9 limbs (2 bilateral), good in 8 limbs (2 bilateral), and fair in 1 case. The differential diagnosis of sural nerve entrapment in athletes is discussed. Increase in sural muscle mass or development of local fibrous scar tissue compromised the sural nerve in its course through the unyielding and inextensible superficial sural aponeurosis. PMID- 11032225 TI - Arthroscopic treatment of lateral meniscal cysts using an outside-in technique. AB - This paper describes 19 cases of lateral meniscal cysts treated arthroscopically using an outside-in technique. In all patients, a horizontal or radial lesion (or both) of the meniscus was present. After the meniscal lesion was arthroscopically removed, the cyst was decompressed both from inside and percutaneously from outside with a motorized instrument introduced through a transmeniscal approach. The follow-up ranged from 2 to 5 years with an average of 3.3 years. On the basis of the evaluation scale developed by Cerullo et al. (1991), the results were rated excellent or good in 17 patients and fair in 2. No patient had cyst recurrence. The computed tomography follow-up examination, done in 10 of the 19 patients after an average of 3 years, showed a good remodeling of the meniscus. One of the two patients with a fair result had hypermobility of the posterior horn of the lateral meniscus due to the resection of the meniscal tissue overlying the popliteus tendon, whereas the other had pain and patellar crepitation owing to a preexisting patellofemoral abnormality. PMID- 11032226 TI - Open versus closed kinetic chain exercises for patellofemoral pain. A prospective, randomized study. AB - The goal of this prospective study was to evaluate the efficacy of open versus closed kinetic chain exercises in the nonoperative management of patellofemoral pain. Sixty patients were randomized into a 5-week program that consisted of only closed kinetic chain exercises or only open kinetic chain exercises. Muscle characteristics, subjective symptoms, and functional performance were evaluated in this study at the time of the initial physical examination, at the end of the treatment period, and 3 months later. Both groups experienced a statistically significant decrease in pain and an increase in functional performance. This study shows that both open and closed kinetic chain exercise programs lead to an improved subjective and clinical outcome in patients with anterior knee pain. The few significantly better functional results for some of the tested parameters in the closed kinetic chain group suggest that this type of treatment is a little more effective than the open kinetic chain program in the treatment of these patients. PMID- 11032227 TI - The influence of reverse-thread screw femoral fixation on laxity measurements after anterior cruciate ligament reconstruction with hamstring tendon. AB - In arthroscopically assisted anterior cruciate ligament reconstruction using hamstring tendon graft, the graft rotates slightly as the femoral screw is inserted. Its final position tends to be in the anterior half of the tunnel in right knees, resulting in clinical laxity. To perform identical procedures on left and right knees, a reverse-thread screw was designed for femoral fixation in right knees. We prospectively studied 80 patients undergoing right-knee anterior cruciate ligament reconstruction with hamstring tendon autograft. Thirty-six patients underwent reconstruction with a standard screw and 44 underwent reconstruction with a reverse-thread screw. The same technique, performed by the same surgeon, was used on all patients. At 12 months' follow-up, the average side to-side differences on arthrometry testing were 2.00 mm for the standard screw group and 0.95 mm for the reverse-thread screw group using a manual maximum test, and 1.66 mm and 1.00 mm, respectively, using the 20-pound test. Both differences were statistically significant. Of the standard group, 23% had a manual maximum difference of 3 mm or more, compared with 8% of the reverse-thread group. A significant difference was found between these two groups for Lachman test (77% with grade 0 for the standard group compared with 92% for the reverse group) but pivot shift and Lysholm knee score were not significantly different. The use of a reverse-thread screw for femoral fixation in right-knee anterior cruciate ligament reconstructions in men significantly decreased laxity at 12 months after surgery compared with standard screw fixation. PMID- 11032228 TI - Change in meniscal strain with anterior cruciate ligament injury and after reconstruction. AB - Meniscal injury has been well documented in association with injury to the anterior cruciate ligament. The purpose of this study was to evaluate the effect of anterior cruciate ligament transection and reconstruction on meniscal strain. Four differential variable reluctance transducer strain gauges were placed in the medial and lateral menisci of nine cadaveric knees. Each specimen was mounted to a six-degree-of-freedom knee testing device. Testing was conducted with the knee fully extended and at 45 degrees and 90 degrees of flexion, both with and without applied axial load. At each angle of flexion, an anterior and posterior tibial load was applied. Next, the anterior cruciate ligament was transected and the testing sequence was repeated. Finally, the ligament was reconstructed using a central one-third patellar tendon graft and the testing sequence was repeated. The results demonstrated statistically significant increases in meniscal strain in ligament-transected knees compared with intact specimens. A reduction in meniscal strain to a level similar to that detected in the ligament-intact knees was observed after anterior cruciate ligament reconstruction. These results have important clinical implications regarding the potentially deleterious effect of the anterior cruciate ligament-deficient knee on meniscal strain and the potential benefit of anterior cruciate ligament reconstruction. PMID- 11032229 TI - Interference screw fixation strength of a quadrupled hamstring tendon graft is directly related to bone mineral density and insertion torque. AB - The purpose of this study was to determine whether bone mineral density of the host bone, measured using conventional dual photon absorptiometry techniques, and insertion torque can predict part of the ultimate failure strength of interference screw fixation of quadrupled hamstring tendon grafts. The semitendinosus and gracilis tendons were harvested from 10 human cadaveric knees, mean age 66.5 years (range, 53 to 81). The bone tunnel was sized within 0.5 mm of the graft. The graft was fixed with a biodegradable screw (7 x 25 mm for the femur, and 9 x 25 mm for the tibia) directly against the tendon and at the joint surfaces. Tibial fixation and femoral fixation were tested to failure using a materials testing system. Bone mineral density was measured in the metaphyseal region of the tibia and femur. The results of multiple regression analyses showed that both insertion torque and bone mineral density were related to the maximum load the graft withstood. These two variables explained 77.1% of the maximum load observed. We concluded that bone mineral density measurements of the host bone site are an important determinant of postoperative graft strength and thus have an important, but previously unrecognized, clinical role in establishing individual postsurgery rehabilitation protocols. Insertion torque in this study was a useful predictor of graft fixation strength. PMID- 11032230 TI - The effect of monopolar radiofrequency treatment pattern on joint capsular healing. In vitro and in vivo studies using an ovine model. AB - The purpose of this study was to compare joint capsular healing after two delivery patterns of monopolar radiofrequency energy: 1) uniform treatment of the joint capsule (paintbrush pattern) and 2) multiple single linear passes (grid pattern). First, an in vitro study was performed to compare the percent shrinkage of these two treatment patterns using the femoropatellar joints (stifles) of six sheep. Monopolar radiofrequency energy (settings, 70 degrees C/15W) was applied to the lateral joint capsule; the treated area was approximately 10 x 10 mm. There was no significant difference in shrinkage between the grid (27% +/- 8.7%) and paintbrush (29% +/- 7.9%) patterns. In the in vivo study, stifles of 24 sheep were randomly assigned to the paintbrush or the grid pattern groups and treatment was performed arthroscopically. Sheep were sacrificed immediately after surgery, or at 2, 6, or 12 weeks after surgery. At 6 weeks after surgery, confocal microscopy demonstrated that treated areas had almost completely repaired in the grid group; some nonviable areas were still present in the paintbrush group. Mechanical testing at 6 weeks indicated that joint capsule in the grid group had better mechanical properties than capsule in the paintbrush group. This study revealed that radiofrequency treatment of joint capsule in a grid pattern allowed faster healing than tissue treated in a paintbrush pattern. PMID- 11032231 TI - Radiofrequency energy-induced heating of bovine articular cartilage using a bipolar radiofrequency electrode. AB - Radiofrequency energy is used for thermal-assisted chondroplasty to treat grade II and III chondromalacia with the lowest possible energy setting that achieves the desired result. The purpose of this in vitro study was to determine the temperature changes associated with the use of radiofrequency energy delivered at different settings to bovine articular cartilage using a bipolar electrode. Cartilage samples were placed in a temperature-controlled (37 degrees C) saline bath for the delivery of radiofrequency energy. A fluoroptic thermometry probe was positioned to record the temperatures at the electrode-tissue interface. The electrode was activated for 2 seconds at settings of V2-120, V2-60, V2-40, and V2 20 in two modes: ablation and desiccation. Additionally, the cartilage samples were visually inspected to determine changes in appearance. The highest average temperatures were as follows: ablation mode, 78.5 degrees C (V2-120), 62.6 degrees C (V2-60), 58.1 degrees C (V2-40), and 54.1 degrees C (V2-20); desiccation mode, 71.8 degrees C (V2-120), 61.4 degrees C (V2-60), 57.7 degrees C (V2-40), and 53.3 degrees C (V2-20). There were statistically significant increases in temperatures associated with each of the respective settings. There were no substantial visual changes produced by the V2-20 settings, while the other settings produced a gradation of effects. These data provide information to help guide the use of a bipolar radiofrequency electrode and electrosurgical system for thermal-assisted chondroplasty. PMID- 11032232 TI - Immunohistochemical analysis for neural markers of the lateral retinaculum in patients with isolated symptomatic patellofemoral malalignment. A neuroanatomic basis for anterior knee pain in the active young patient. AB - We evaluated 13 lateral retinacula excised at the time of Insall proximal realignments or isolated lateral retinacular releases performed in patients with isolated symptomatic patellofemoral malalignment recalcitrant to nonoperative treatment. Evaluation was performed by means of conventional histologic and immunohistochemical analysis for neural markers (S-100 protein, neurofilament protein, substance P, and neural growth factor). The observations reported here provide a neuroanatomic basis for anterior knee pain syndrome in active young patients with isolated symptomatic patellofemoral malalignment and support the clinical observation that the lateral retinaculum may have a key role in the origin of this pain as a result of increased neural growth factor production, which induces proliferation of nociceptive axons, mainly in a perivascular location. PMID- 11032233 TI - Refracture of proximal fifth metatarsal (Jones) fracture after intramedullary screw fixation in athletes. AB - This study details six instances of refracture of clinically and radiographically healed fractures of the base of the fifth metatarsal after intramedullary screw fixation. Four professional football players, one college basketball player, and one recreational athlete underwent intramedullary screw fixation of fifth metatarsal fractures. The athletes were released to full activities an average of 8.5 weeks (range, 5.5 to 12) after fixation, when healing was clinically and radiographically documented. Three football players developed refracture within 1 day of return to full activity. The other three athletes refractured at 2.5, 4, and 4.5 months after return to activity. Two football players underwent repeat fixation with larger screws and returned to play in the same season. The college basketball player underwent bone grafting and returned to play in subsequent seasons. The other three athletes underwent nonoperative management and healed uneventfully over 6 to 8 weeks. On the basis of this series, we recommend that 1) screw fixation using a large-diameter screw should be given careful consideration for patients with large body mass for whom early return to activity is important; 2) functional bracing, shoe modification, or an orthosis should be considered for return to play; 3) if refracture occurs, exchange to a larger screw may allow return to play in the same season; and 4) alternative imaging should be considered to help document complete healing. PMID- 11032234 TI - Repair of a bucket-handle tear of a complete discoid lateral meniscal incarcerated in the posterolateral compartment. PMID- 11032235 TI - Hypothenar hammer syndrome in a golf player. A case report. PMID- 11032236 TI - Ulnar periostitis due to stress in hockey players. PMID- 11032237 TI - Management of unicompartmental arthritis in the anterior cruciate ligament deficient knee. AB - There exists a group of patients who are difficult to manage because they have both anterior knee instability secondary to anterior cruciate ligament deficiency and unilateral degenerative joint disease. A large majority of these patients report a history of previous meniscal injury or meniscectomy after knee trauma at a relatively young age. Active patients who report symptomatic knee instability or pain associated with athletics or activities of daily living after conservative treatment may be indicated for surgery. Current endoscopic methods of anterior cruciate ligament reconstruction result in low patient morbidity, the elimination of anterior knee instability, and a timely return of function. Osteotomies about the knee joint are an effective means of treating unicompartmental knee arthrosis. Long-term studies have demonstrated that knee osteotomy is a good surgical option for patients with unicompartmental arthritis who are considered too young for total knee arthroplasty. We describe a comprehensive treatment approach to the patient with anterior cruciate ligament deficiency and isolated medial or lateral osteoarthritis. An assessment of pain symptoms, instability symptoms, and lower extremity alignment is used to formulate a treatment plan. PMID- 11032238 TI - Graft fixation in cruciate ligament reconstruction. AB - Cruciate ligament reconstruction has progressed dramatically in the last 20 years. Anatomic placement of ligament substitutes has fostered rehabilitation efforts that stress immediate and full range of motion, immediate weightbearing, neuromuscular strength and coordination, and early return to athletic competition (3 months). This has placed extreme importance on secure graft fixation at the time of ligament reconstruction. Current ligament substitutes require a bony or soft tissue component to be fixed within a bone tunnel or on the periosteum at a distance from the normal ligament attachment site. Fixation devices have progressed from metal to biodegradable and from far to near-normal native ligament attachment sites. Ideally, the biomechanical properties of the entire graft construct would approach those of the native ligament and facilitate biologic incorporation of the graft. Fixation should be done at the normal anatomic attachment site of the native ligament (aperture fixation) and, over time, allow the biologic return of the histologic transition zone from ligament to fibrocartilage, to calcified fibrocartilage, to bone. The purpose of this article is to review current fixation devices and techniques in cruciate ligament surgery. PMID- 11032239 TI - Chromatin remodeling and T helper subset differentiation. AB - The T helper subsets Th1 and Th2 regulate specific types of immune responses by producing distinct sets of cytokines. Differentiation of the T helper subsets from their common precursors, naive CD4+ T cells, is induced by antigen stimulation and controlled by various other conditions. Of these conditions, the contributions of the cytokine environment have been the best characterized. The presence of interleukin-4 (IL-4) directs the differentiation towards Th2 cells, whereas IL-12 induces Thl differentiation. The Th2 signature cytokine genes encoding IL-4, IL-13, and IL-5 are clustered, and noncoding regions such as the intergenic region of the IL-4 and IL-13 genes are highly conserved from mice to humans. Alteration of the chromatin structure of this Th2 cytokine cluster region is detected as nuclease-accessible regions specific to Th2 cells. Activation of STAT6 promotes Th2 differentiation by inducing Th2-specific transcription factors, including GATA3. Expression of GATA3 induces Th2 differentiation and enhances the Th2 cell-specific chromatin accessibility, indicating that GATA3 is a key protein that regulates differentiation through chromatin remodeling. T helper subset differentiation provides a good system to study gene expression regulation at the chromatin level. PMID- 11032240 TI - Hereditary degenerative retinopathies: optimism for somatic gene therapy. AB - Retinitis pigmentosa comprises a large and exceptionally heterogeneous group of hereditary disorders of the retina. As a result of an extensive investigation around the world, primary genetic lesions have been described in many genes. Some of these genes encode enzymes that are involved in the signal transduction pathway. On the basis of in vitro functional assays and standard transgenic and knock-out experiments, it has been proposed that normal cell functions are disrupted because of an abnormal protein-folding and metabolic errors or structural defects in the membrane. This ultimately leads to a gene-mediated cell death known as apoptosis. Various gene transfer approaches using mouse models further suggest that the degeneration can be rescued to some extent. Although many questions remain to be answered, investigations during the last 10 years have enormously increased our understanding of this exceptionally heterogeneous disorder and give hope for an effective gene therapy and a possible cure. PMID- 11032241 TI - The presence of a glycosyl phosphatidylinositol-anchored alpha-mannosidase in boar sperm. AB - alpha-Mannosidase and beta-galactosidase were released from boar sperm into the medium by treatment with calcium ionophore A23187 or by 0.2% Brij-35/2% acetic acid. About half as much alpha-mannosidase activity as that in the acid extract was recovered by digestion with phosphatidylinositol-specific phospholipase C (PI PLC), whereas the liberation rate of beta-galactosidase treated with PI-PLC was low. These results suggest that some alpha-mannosidase is anchored in the plasma membrane of the acrosomal region by attachment to the lipid phosphatidylinositol and that beta-galactosidase is localized mainly in the acrosome or integrated in the plasma membrane by a spanning stretch of hydrophobic peptides. beta Galactosidase, which is present as an oligomers in the acid extract of sperm, dissociated into monomers under weakly alkaline conditions; under acidic conditions, the monomers associated again. No pH-sensitive association dissociation of alpha-mannosidase was observed. PMID- 11032242 TI - Effect of high concentrations of sucrose on the enzymatic activity of alpha chymotrypsin. AB - The effect of the low-molecular-mass natural reagents in high concentrations is important for investigating enzymatic reactions in near "in vivo" conditions and for optimisation of biotechnology processes. A model system, including p nitrophenyl acetate as substrate and alpha-chymotrypsin as proteolytic enzyme, has been used to study the effect of high concentrations of sucrose, both influencing the viscosity of the reaction medium and acting as a nucleophilic effector (activator) on the enzymatic reaction. A kinetic scheme at high concentrations of nucleophilic effectors (sucrose) has been proposed. PMID- 11032243 TI - Age-related changes of liver antioxidant enzymes and 8-hydroxy-2'-deoxyguanosine during fetal-neonate transition and early rat development. AB - We have studied the pro-antioxidant status of the rat liver on the last day of gestation and at 1, 15, and 30 days of extrauterine life. Representative variables, such as activities of superoxide dismutase (SOD), catalase, and glutathione peroxidase and concentrations of reduced glutathione and 8-hydroxy-2' deoxyguanosine, were determined in liver to assess the degree of birth-associated oxidative stress during the fetal-neonatal transition and early development of the rat. Percentages by which liver Cu/ZnSOD activity increased over the basal value of the fetal liver were 54%, 95%, and 127% at neonatal days 1, 15, and 30, respectively. There was a lack of induction in the development profile of MnSOD. Catalase activity was clearly and progressively induced with time from the fetal state up to the neonatal age of 1 month. Glutathione peroxidase activity and glutathione content showed a tendency to decline during the first day after birth, though they increased to significantly higher values on days 15 and 30. However, the amount of rat liver 8-hydroxy-2'-deoxyguanosine did not increase. These results suggest that the induced antioxidant activities may be responsible for maintaining DNA stability during the perinatal development of the rat liver. PMID- 11032244 TI - Regulation of cardiac Jak-2 in animal models of insulin resistance. AB - Insulin induces phosphorylation and activation of JAK2 tyrosine, as well as its association with STAT1 and SHP2 in insulin-sensitive tissues of intact rats, thus demonstrating a new pathway in transduction of insulin signals. We investigated this pathway in hearts of rats in three situations of insulin resistance: 72 h of fasting, chronic treatment with dexamethasone, and acute treatment with epinephrine. The acute treatment with epinephrine showed no difference in insulin induced JAK2 tyrosine phosphorylation or JAK2/STAT1 and JAK2/SHP2 association in comparison with the control. In fasted rats the JAK2 protein concentration decreased, accompanied by a decrease in the stoichiometry of the phosphorylation to 70%, an increase in association of JAK2/STAT1 to 160%, and a decrease in JAK2/SHP2 association to 85%. In the dexamethasone-treated group, the JAK2 protein concentrations increased but the stoichiometry of its phosphorylation decreased to 20%, whereas the JAK2/STAT1 and JAK2/SHP2 associations changed by 70% and 170%, respectively. In fasting and dexamethasone-treated rats, therefore, insulin-induced JAK2 tyrosine phosphorylation decreases, and the JAK2 protein expression is differentially regulated such that the insulin-induced JAK2 association with SHP2 and STAT1 shows opposite interactions with the kinase. PMID- 11032245 TI - The protein composition of the hepatocyte nuclear matrix is differentiation-stage specific. AB - The protein composition of hepatocyte nuclear matrices was examined in rats from the 16th day of gestation to 75 days after birth (adult). An overall increase in size of the nuclear matrix was accompanied by quantitative and qualitative changes in its protein content. Quantitative changes of the major proteins of the peripheral lamina surrounding the isolated nuclear matrix were detected. By Western analysis we established that in pre- and postnatal nuclear matrices the relative concentrations of lamin C were greater than lamin A. After birth, the relative concentrations of both lamins progressively increased. In the adult nuclear matrix, the concentration of lamin A was greater than lamin C. In contrast, the relative concentrations of lamin B remained unchanged throughout development and growth. The relative concentrations of two nuclear matrix associated regulatory proteins studied changed with development and growth: transcription factor C/EBPalpha isoforms, which were detected during the gestation period, increased notably after the first postnatal day, attaining a maximum at the adult stage; the high concentrations of the proliferating cell nuclear antigen (PCNA) perceptibly decreased after the 21st prenatal day. Changes in the composition of the nuclear matrix protein suggest that this structure coordinates nuclear functioning during cell differentiation. PMID- 11032246 TI - Differential effects of tri-n-butylstannyl benzoates on induction of apoptosis in K562 and MCF-7 cells. AB - Tributyltin compounds have been shown to induce apoptosis by causing extracellular Ca2+ influx and generating reactive oxygen species (ROS). Several organotin compounds were reported to have differential cytotoxicity on various human cell lines depending on the length of the alkyl chain. In this report, the cytotoxic effects of three tri-n-butylstannyl (halo)benzoate compounds-tri-n butylstannyl benzoate (TBSB), tri-n-butylstannyl-2,6-difluorobenzoate (TBSDFB) and tri-n-butylstannyl-2-iodobenzoate (TBSIB)-were studied on lymphocytic cells of human leukemic K562 lineage and epithelial cells of human breast cancer MCF-7 cells lineage. K562 cells were found to be more sensitive to these compounds than MCF-7 cells. Although the induction of apoptosis by the above compounds in K562 cells resulted from the extracellular Ca2+ influx and the generation of ROS, the initial amount of extracellular Ca2+ influx was greater in TBSB-treated K562 cells than the cells treated with either TBSDFB or TBSIB. Similarly, DNA fragmentation by endonucleases was observed as an early event in TBSB-treated K562 cells, which might be correlated with the initially greater extracellular Ca2+ influx. In contrast, MCF-7 cells were found to undergo apoptosis mainly because of the generation of ROS. The present results suggest that the differential effects of tributyltin compounds on induction of apoptosis in K562 and MCF-7 cells are largely attributable to the extent of extracellular Ca2+ influx. PMID- 11032247 TI - The nitrogen source-dependent 126-kDa protein from Synechococcus PCC 7942 plasmalemma: a trimer of the NrtA nitrate-binding protein. AB - Expression of a 126-kDa protein in the plasmalemma (cytoplasmic membrane) from Synechococcus PCC 7942 is dependent on the nitrogen source. Polyclonal antibody raised against the NrtA protein reacted with the 126-kDa protein. Two peptide sequences from the 126-kDa protein were retrieved in NrtA. FPLC purification of plasmalemma solubilised in Triton X-100 gave a fraction consisting mainly of a 126-kDa component (72.6%), as shown by sedimentation velocity. Equilibrium sedimentation of the same fraction gave evidence of the existence of an oligomeric structure (Ka = 2 x 10(3) and 2.9 subunits). Thus, the 126-kDa protein is considered as a trimeric arrangement of the 45-kDa protein in the plasmalemma. PMID- 11032248 TI - Maesaquinone: a novel inhibitor of plant mitochondrial respiratory enzymes that react with ubiquinone. AB - The effect of maesaquinone, 2-(14-nonadecenyl)-3,6-dihydroxy-5-methyl-1,4 benzoquinone, on plant mitochondrial respiration has been investigated. In mitochondria isolated from thermogenic Arum maculatum spadices, this compound inhibits both cytochrome and alternative pathway activities. Kinetic analyses reveal that this inhibition is the result of potent effects of maesaquinone on the alternative oxidase (ID50 < 0.3 microM) and complex III (ID50 < 5 microM). Succinate dehydrogenase and external NADH dehydrogenase are also inhibited, albeit to a lesser extent (approximately 30% at 1 microM). These data suggest that maesaquinone specifically affects the interaction of the respective enzymes with ubiquinone. PMID- 11032250 TI - Selection of peptide ligands that bind to acid fibroblast growth factor. AB - Acid fibroblast growth factor (aFGF) binds to its cell-surface receptors in a heparin-dependent manner. In an attempt to define the aFGF recognition site on fibroblast growth factor receptor 1 (FGFR1), we developed a screening strategy for identifying FGF ligands that bind to the receptor-binding region of FGF. To retain the natural conformation of aFGF during screening, we used biotinylated heparin to immobilize aFGF on a streptavidin-coated dish. A 15-mer phage display peptide library was then screened in the dish and a group of related peptide sequences was identified. These peptide sequences contain two conserved motifs, SSG and VPS, corresponding to two protein sequences of the immunoglobulin-like (Ig-like) domain II of FGFR1 at amino acids 180-182 and 221-223 (CPSSG VPSDKGNYTC). Further experiments demonstrate that the phage displaying these sequences can specifically bind to aFGF and that the synthesized peptide corresponding in sequence can inhibit mitogenic activity of aFGF. These sequences may thus constitute part of the aFGF-binding region on FGFR1, and the synthesized peptide has the potential to become a therapeutic agent as an aFGF antagonist. PMID- 11032249 TI - Mechanism of free radical production in exhaustive exercise in humans and rats; role of xanthine oxidase and protection by allopurinol. AB - Exhaustive exercise generates free radicals. However, the source of this oxidative damage remains controversial. The aim of this paper was to study further the mechanism of exercise-induced production of free radicals. Testing the hypothesis that xanthine oxidase contributes to the production of free radicals during exercise, we found not only that exercise caused an increase in blood xanthine oxidase activity in rats but also that inhibiting xanthine oxidase with allopurinol prevented exercise-induced oxidation of glutathione in both rats and in humans. Furthermore, inhibiting xanthine oxidase prevented the increases in the plasma activity of cytosolic enzymes (lactate dehydrogenase, aspartate aminotransferase, and creatine kinase) seen after exhaustive exercise. Our results provide evidence that xanthine oxidase is responsible for the free radical production and tissue damage during exhaustive exercise. These findings also suggest that mitochondria play a minor role as a source of free radicals during exhaustive physical exercise. PMID- 11032251 TI - Transport of mitochondria during axonogenesis. AB - The cellular mechanisms involved in axonogenesis are still unclear. In the present work we found that formation of neurites in cultured neonatal dorsal root ganglion neurons co-incided with the redistribution of highly charged mitochondria. Radially distributed in subplasmalemmal space 3 h after plating, highly charged mitochondria formed clusters in the hillocks of predominant neurites during the next 2448 h and then redistributed into the axons. These results provide evidence that accumulation of a critical mass of charged mitochondria at the site of the future axonal hillock may represent the slow initiation stage of axonogenesis, followed by a fast growth phase. PMID- 11032252 TI - Safety, efficacy, and durability of carotid artery stenting for restenosis following carotid endarterectomy: a multicenter study. AB - PURPOSE: To present the results of a multicenter registry established to collect data on carotid stent procedures in patients with restenosis following carotid endarterectomy. METHODS: The procedural details, outcomes, and late follow-up results were collected from 14 centers in the United States. Thirty-day and late stroke and death rates were analyzed. RESULTS: Three hundred and thirty-eight patients (201 men; 71 +/- 8 years) underwent carotid stenting in 358 arteries. The average duration from carotid endarterectomy was 5.5 +/- 7.3 years. Sixty-one percent of the patients were asymptomatic. The overall 30-day stroke and death rate was 3.7%. The minor stroke rate was 1.7% (6/358), and the major nonfatal stroke rate was 0.8% (3/358). The fatal stroke rate was 0.3% (1/358), and the nonstroke-related death rate was 0.9% (3/338). There was 1 (0.3%) fatal and 1 (0.3%) nonfatal stroke during the follow-up period. The overall 3-year rate of freedom from all fatal and nonfatal strokes was 96% +/- 1% (+/- SE). CONCLUSIONS: Carotid artery stenting can be performed in patients with restenosis following carotid endarterectomy with 30-day complication rates comparable to those of most published studies on repeat carotid endarterectomy. Results of late follow-up suggest that this technique is durable and efficacious. PMID- 11032253 TI - Endovascular repair of an extracranial internal carotid artery aneurysm complicated by heparin-induced thrombocytopenia and thrombosis. AB - PURPOSE: To report the endovascular treatment of a symptomatic extracranial internal carotid artery (ICA) aneurysm that was complicated by heparin-induced thrombocytopenia and thrombosis. METHODS AND RESULTS: After undergoing a coronary artery bypass graft procedure, a patient was diagnosed with a symptomatic, 3.5-cm ICA aneurysm by computed tomography and angiography. Via a semiclosed access, an Enduring vascular graft was inserted under controlled back bleeding from the ICA. The patient was recovering uneventfully when routine duplex scanning on the fifth postoperative day suggested multiple thrombi within the graft, which was confirmed by arteriography. Thrombectomy and local fibrinolysis were performed; however, the graft occluded the next day without causing neurological symptoms. Heparin-induced thrombocytopenia was diagnosed by enzyme-linked immunosorbent assay. CONCLUSIONS: Endovascular repair of high cervical extracranial ICA aneurysms is feasible, and protection against intracerebral embolization can be achieved using a semiclosed technique with controlled back bleeding from the ICA during endograft deployment. However, multiple thrombi or thrombotic occlusion during the postoperative period, particularly in a patient already sensitized to heparin, should direct attention toward possible heparin-induced thrombocytopenia. PMID- 11032254 TI - Endovascular AAA repair attenuates the inflammatory and renal responses associated with conventional surgery. AB - PURPOSE: To quantify the inflammatory and renal parameters in comparative cohorts of patients undergoing surgical or endovascular repair of abdominal aortic aneurysms (AAAs). METHODS: Forty-three patients (41 men; ages 58-81 years) underwent endovascular or conventional aneurysm surgery according to aortic morphology. All patients received a standard general anesthetic and had 12 serial blood and urine samples collected during the perioperative period. Samples underwent analysis for the cytokines interleukin (IL) 1beta tumor necrosis factor alpha (TNF-alpha), and IL-6. White cell and platelet activation were estimated indirectly by measuring sL-selectin and 11-dehydrothromboxane B2, respectively. The urinary albumin:creatinine ratio (ACR) and N-acetyl-beta-D-glucosaminidase (NAG) activity were estimated to assess renal injury. Fibrinogen and fibrinogen degradation products were calculated to assess activation of the clotting cascade. RESULTS: Twenty-three patients underwent endovascular AAA repair and 20 had conventional surgery. Concentrations of IL-6 (p < 0.002) and TNF-alpha (p < 0.0004) were significantly higher in the conventional group. The ACR (p < 0.002) and urinary NAGs (p < 0.0009) were also significantly higher in this group, suggesting greater renal injury. Platelet activity was significantly greater in the endovascular group (p < 0.01), perhaps indicating thrombus organization within the aneurysm sac. CONCLUSIONS: These data suggest that the inflammatory response associated with conventional aneurysm repair is largely obviated by endovascular techniques. This may potentially translate to a lower incidence of multiple organ failure after endovascular surgery. PMID- 11032255 TI - Quality of life before and after endovascular and open repair of asymptomatic AAAs: a prospective study. AB - PURPOSE: To assess how patients perceive health-related quality of life (HRQOL) after endovascular and open abdominal aortic aneurysm (AAA) repair. METHODS: Forty-two consecutive patients (33 men; mean age 74 years, range 46-81) were assessed prospectively before and after elective endovascular (n = 21) and open (n = 21) AAA repair. Aneurysm morphology dictated the type of repair. The two patient groups were similar regarding age, gender, comorbidities, and cardiopulmonary function. Data concerning surgical trauma were compiled. The Nottingham Health Profile (NHP) score was used to assess the perceived HRQOL (criteria: pain, mobility, sleep, emotion, energy, and isolation) preoperatively and at 5, 30, and 90 days postoperatively. Specific treatment perception questions were added. RESULTS: One patient from each group died, leaving 40 patients to complete the study. Two patients with open repair and 1 patient with endovascular repair were unfit to answer the questionnaire on day 5. The HRQOL improved at 3 months compared with the preoperative values (p < 0.05). No significant difference was found at any time between the open and endovascular groups regarding the NHP score, although the operative time, blood loss, analgesic use, and hospital stay were significantly in favor of endovascular repair. Reinterventions were required in 5 patients with endovascular and 2 patients with open repair. CONCLUSIONS: In general, 3 months after AAA repair, the perceived HRQOL seems better than before treatment. Perceived HRQOL is similar after endovascular and open AAA repair despite greater surgical trauma in open surgery. This may reflect the higher number of reinterventions following endovascular repair but also difficulties in defining HRQOL. PMID- 11032256 TI - Three-dimensional intravascular ultrasound assessment of abdominal aortic aneurysm necks. AB - PURPOSE: To document the accuracy of an automated analysis system for measuring lumen diameter and neck lengths of abdominal aortic aneurysms (AAAs) from intravascular ultrasound (IVUS) images and to describe additional features associated with 3-dimensional (3D) IVUS imaging. METHODS: Twenty-two aortic aneurysms were studied with IVUS. Lumen diameters obtained using the automated analysis system were compared with manual measurements from axial IVUS scans, as were neck lengths obtained using automated analysis versus those measured with the aid of a displacement sensing device. Automated analyses were repeated by a second observer. Agreement was expressed as the coefficient of variation (CV). RESULTS: Twenty proximal aortic, 6 distal aortic, and 3 iliac necks were available for analysis. Comparison between automated analysis and manual measurements for lumen diameter revealed a difference of 0.45 +/- 0.42 mm (mean +/- SD, Pearson's r = 0.99, p < 0.001, CV = 2.1%) and a difference of 0.05 +/- 0.12 cm (r = 0.99, p = 0.04, CV = 4.1%) for neck length. Interobserver difference for lumen diameter was 0.13 +/- 0.66 mm (r = 0.99, p < 0.001, CV = 3.4%) and 0.05 +/- 0.11 cm for length measurements (r = 0.99, p = 0.02, CV = 3.5%). The 3D IVUS imaging facilitated the identification of neck configuration. CONCLUSIONS: Automated analysis of IVUS images allows accurate measurement of the lumen diameter of proximal and distal AAA necks and gives length measurements comparable to those of manual analysis. Longitudinal display of IVUS images aids in the elucidation of neck anatomy. PMID- 11032257 TI - A simple technique to improve the accuracy of proximal AAA endograft deployment. AB - PURPOSE: To report a technique for overcoming the positioning errors caused by angulation and rotation of the proximal aortic neck when anteroposterior fluoroscopic imaging is used during endograft deployment. TECHNIQUE: Aortic neck angulation and rotation were measured preoperatively using spiral computed tomographic angiography in sagittal and axial projections. Before proximal graft deployment, the proximal end of the endograft was centered in the field of view, and the position of the C-arm was adjusted to the aortic neck angulation. Using this technique, optimal positioning of the endograft relative to the true position of the renal arteries can be achieved. CONCLUSIONS: C-arm angulation and rotation is helpful in facilitating perfect positioning for an optimal seal between the endograft and the infrarenal aortic neck. PMID- 11032258 TI - Popliteal artery aneurysms: is endovascular reconstruction durable? AB - PURPOSE: To describe an endovascular method of performing femoropopliteal in situ saphenous vein (SV) bypass and popliteal artery aneurysm (PAA) embolization. METHODS: Twenty-two patients underwent PAA operations. Twelve patients had conventional SV bypasses with PAA proximal and distal ligation, whereas 10 underwent PAA embolization and an endovascular in situ SV bypass (EISB). The endovascular procedure was performed using an angioscopically guided side branch coil occlusion system. The PAAs were coil embolized under fluoroscopic surveillance. RESULTS: No deaths or wound complications occurred in the EISB group. The mean hospital length of stay (LOS) was 2.1 days. Seven EISB procedures were performed through 2 incisions, whereas 3 operations required an additional incision. One graft occluded at 3 months. All PAAs remained occluded by color flow ultrasonography at follow-up ranging from 4 to 23 months (mean 13.6); cumulative primary patency was 89%. In the conventional bypass group, no deaths occurred, but 3 (25%) patients had wound complications. The mean LOS was 6.2 days, and 1 graft occluded, giving an 86% cumulative primary patency at 42 months. CONCLUSIONS: This minimally invasive technique obviates an extensive incision to harvest the SV and ligate the PAA proximally and distally. If long term endovascular bypass graft patency and PAA occlusion rates prove to be similar to open operative results, the benefits of reduced wound complications, decreased hospital LOS, and increased health care savings support further investigation of this endovascular approach for the treatment of PAA. PMID- 11032259 TI - Acute renal failure after redo thoracoabdominal aortic aneurysm repair in a patient with a solitary kidney: successful percutaneous treatment. AB - PURPOSE: To report the successful percutaneous treatment of renal artery stenosis that precipitated renal failure following surgical repair of a thoracoabdominal aortic aneurysm (TAAA). METHODS AND RESULTS: A 70-year-old woman with a solitary kidney became anuric 2 hours after urgent repair of a symptomatic true aneurysm of the Carrel patch from an 8-year-old TAAA repair. After medical treatment failed, aortography was performed, identifying complete occlusion of the solitary renal artery. Balloon dilation and implantation of a Palmaz stent restored renal perfusion and improved function. At 6-month follow-up, she was normotensive and her creatinine within normal limits. CONCLUSIONS: Renal artery stenosis or occlusion is a treatable cause of acute renal failure after TAAA repair. Percutaneous treatment options are likely to be better tolerated than surgical revascularization in this patient population. PMID- 11032260 TI - Suprarenal mycotic aneurysm exclusion using a stent with a partial autologous covering. AB - PURPOSE: To report a combined endovascular and open technique to manage a suprarenal mycotic aortic aneurysm using a stent-graft partially covered with a section of autologous artery. METHODS AND RESULTS: A 50-year-old was hospitalized for staphylococcal septicemia and severe back pain. A previously diagnosed 3-cm abdominal aortic aneurysm was found to have expanded 2 cm in 3 weeks. Aortography documented some periaortic thickening and 2 mycotic aneurysms, one posterior at the level of the superior mesenteric artery and the second at the aortic bifurcation. After intensive antibiotic therapy, an endovascular approach to exclude the suprarenal mycotic aneurysm was undertaken in tandem with surgical excision of the infrarenal aneurysm. The harvested right common iliac artery was used to partially cover a Palmaz stent, which was deployed under direct vision just above the renal artery ostia so that the covered portion of the stent excluded the aneurysm. A right axillofemoral bypass with a femorofemoral bypass completed the revascularization. Postoperatively, the patient developed renal failure, ischemic colitis necessitating a left hemicolectomy, and paraplegia. Although the patient is paralyzed, the aneurysm remains excluded with patent visceral vessels at 12 months following surgery. No organisms were grown from excised aortic tissue, and no signs of recurrent infection have been seen. CONCLUSIONS: Stent-graft repair may be able to lessen the invasiveness and reduce the morbidity associated with treatment of mycotic aortic aneurysms. PMID- 11032261 TI - Endovascular repair of perisplanchnic abdominal aortic aneurysm with visceral vessel transposition. AB - PURPOSE: To report a combined endoluminal and open surgical approach for a suprarenal abdominal aortic aneurysm (AAA) with coexistent splanchnic vessel stenoses. METHODS AND RESULTS: A 64-year-old man presented with an aneurysm of the proximal abdominal aorta and severe stenoses of the celiac axis and superior mesenteric artery (SMA). An initial 2-stage plan to stent the visceral vessel stenoses and exclude the aneurysm with a fenestrated stent-graft failed when the celiac lesion could not be crossed. The approach was changed to restore visceral perfusion with a bifurcated left iliosplenic and ilio-SMA bypass graft. Exclusion of the aneurysm was achieved with a custom-made suprarenal aortic tube stent graft (Ivancev-Malmo) system. The patient is free of symptoms at 22 months, and there was no aneurysm visible on the 14-month CT scan. CONCLUSIONS: Hybrid techniques are an alternative treatment for complex perivisceral aortic aneurysms when total endovascular reconstruction is not possible. PMID- 11032262 TI - Endovascular management of rare sciatic artery aneurysm. AB - PURPOSE: To present a rare case of complete persistent sciatic artery aneurysm successfully treated by coil embolization and to suggest a new classification for this anomaly that encompasses both clinical and pathological factors. METHODS AND RESULTS: A 77-year-old female presented with sudden onset of sharp, throbbing left thigh and foot pain. A nontender 10.0- x 7.0-cm pulsatile mass was found over the inferolateral aspect of the left buttock on examination in the right lateral decubitus position. Angiography demonstrated a 6.9-cm sciatic artery aneurysm with the distal sciatic artery completely thrombosed. Before planned bypass grafting, the aneurysm and distal segment of the internal iliac artery were successfully embolized with coils through a left percutaneous approach. The patient's symptoms resolved with no deterioration in limb perfusion, and further revascularization was unnecessary. Postembolization imaging showed complete occlusion of the aneurysm, which has continued for 19 months. CONCLUSIONS: The application of endovascular techniques and surgical revascularization procedures provides numerous management options for the treatment of an aneurysmal persistent sciatic artery. This case demonstrates that clinical evaluation between staged procedures may obviate the need for further intervention. PMID- 11032263 TI - Acute interventions for ischemic stroke: present status and future directions. PMID- 11032264 TI - Prevention of side branch endoleaks with thrombogenic sponge: one-year follow-up. PMID- 11032265 TI - Syringe pump assemblies and the natural history of clinical technology. PMID- 11032266 TI - Cumulation and flexibility with infusions of neuromuscular blocking drugs. PMID- 11032267 TI - Cumulation characteristics of cisatracurium and rocuronium during continuous infusion. AB - PURPOSE: The dissimilar pharmacokinetic properties of cisatracurium (CIS) and rocuronium (ROC) predict different potential for drug cumulation when these drugs are administered by continuous infusion. A study was therefore undertaken to compare cumulation potential of CIS and ROC during surgical procedures of relatively long duration (2-4 hr). METHODS: Sufentanil/propofol-N2O anesthesia was administered to 40 ASA I and II adults. In a double-blind protocol, patients were randomly allocated to receive a continuous i.v. infusion of either CIS or ROC, titrated in progressive increments or decrements as required to achieve and maintain 95 +/- 5% depression of the T1 response of the adductor pollicis muscle, using a Datex NMT-100 Relaxograph EMG monitor applied at the wrist. At the end of surgery, 60 microg x kg(-1) neostigmine plus 15 microg x kg(-1) atropine were administered for reversal. RESULTS: The duration of infusion was 104 +/- 33 min in group CIS and 110 +/- 23 min in group ROC (P=NS). In both groups, a progressive decrease in potency-adjusted infusion rates was observed after 30 min, then stabilized beyond 60 min. When allowing for an initial period of stabilization, mean potency-adjusted infusion requirements were: CIS 0.81 +/- 0.02 microg x kg(-1) x min(-1) and ROC 5.58 +/- 1.94 microg x kg(-1) x min(-1). There were no differences between groups at any time with regard to potency adjusted infusion requirements necessary to maintain 90-99% block (P=NS). However, drug costs/hr for maintenance of neuromuscular block were less with CIS ($3.57 +/- 0.09) than with ROC ($6.03 +/- 0.27), P < 0.001. CONCLUSION: When adjusted to equipotency, infusion requirements of CIS and ROC vary at similar rates during general anesthesia. Despite pharmacokinetic differences, neither drug demonstrates cumulation for infusion lasting up to 3.5 hr. PMID- 11032268 TI - Train-of-four fade and neuromuscular block in rats: a comparison between pancuronium, vecuronium, and rocuronium. AB - PURPOSE: To clarify the relationship between neuromuscular block and train-of four fade and to investigate the causes of these drug-dependent differences, we compared the neuromuscular block and TOF fade after pancuronium, vecuronium and rocuronium. METHODS: In 24 anesthetized rats, the sciatic nerve was stimulated, and the twitch of left tibialis anterior muscle was recorded. After T1 (first twitch response) was kept constant at 95% block by administration of pancuronium, vecuronium, or rocuronium (n=8, in each), the TOF fade was measured when T1 block was decreased to 40% and 20%. In addition, using 24 phrenic nerve-diaphragm preparations, the fade was measured when the T1 block increased to 20% and 40% by titrating of either one of the three drugs (n=8, in each). RESULTS: In in vivo experiments, the fade produced by pancuronium was greater than that by vecuronium or rocuronium when T1 block was at 40% (81 +/- 9 vs 63 +/- 15 and 63 +/- 6%, respectively) and at 20% (66 +/- 13 vs 34 +/- 17 and 40 +/- 6%, respectively). In contrast, in in vitro experiments, the differences did not reach significant levels among the three drugs either at 20% (32 +/- 19 vs 33 +/- 10 and 32 +/- 17%) or 40% of block (62 +/- 29 vs 65 +/- 14 and 55 +/- 14%). CONCLUSIONS: For vecuronium and rocuronium, the results were similar in vivo and in vitro. For pancuronium, fade was greater in vivo. These results suggest that different neuromuscular blocking agent have different relationships between the fade and the block. In vitro results might not be the same as in vivo, possibly due to pharmacokinetic differences. PMID- 11032269 TI - Comparison of maternal satisfaction between epidural and spinal anesthesia for elective Cesarean section. AB - PURPOSE: Epidural anesthesia was a commonly used technique for elective Cesarean section. Recently, because of the availability of non-cutting spinal needles, many institutions have changed from epidural to spinal anesthesia. The purpose of this study was to compare maternal satisfaction between epidural and spinal anesthesia for elective Cesarean section with a new satisfaction tool. METHODS: We studied healthy parturients in a randomized, double-blinded pilot study in which patients were assigned to receive either epidural (n = 13) or spinal (n = 14) anesthesia for elective Cesarean section. Two and 24 hr postoperatively, patients completed a validated 22-point maternal satisfaction questionnaire and a 10-cm visual analog score (VAS) for satisfaction. Maternal satisfaction scores were compared between groups. RESULTS: There was no difference in demographics, complications or technical failures between groups. Mean satisfaction scores on the questionnaire (0-154) at two and 24 hr were 130.23 +/- 11.36 and 129.54 +/- 16.70 for the epidural group and 116.92 +/- 18.47 and 115.92 +/- 15.71 for the spinal group (P = 0.04 and P = 0.03 respectively). No difference in VAS scores was noted. The presence of minor side effects including pruritus contributed to the lower satisfaction in the spinal group at 24 hr. CONCLUSION: This pilot study demonstrated higher maternal satisfaction with epidural than with spinal anesthesia for elective Cesarean section. This may be related to the increased side effects caused by neuraxial morphine. The satisfaction questionnaire was able to elucidate differences not detected with a global VAS for satisfaction. Further study with a larger patient population is required to confirm these data. PMID- 11032270 TI - Clonidine prolongs the effect of ropivacaine for axillary brachial plexus blockade. AB - PURPOSE: To evaluate the effect of adding clonidine to ropivacaine, for axillary brachial plexus blockade, on the onset and duration of sensory and motor block and duration of analgesia. METHODS: In a prospective randomised double blind placebo controlled study axillary brachial plexus blockade was performed in 50 patients using 40 ml ropivacaine 0.75%. Group (A) had 150 microg clonidine and Group (B) 1 ml normal saline added to the local anesthetic. Sensory function was tested using pinprick (sharp sensation, blunt sensation or no sensation) and temperature with an ice cube compared with the opposite arm, (cold/not cold). Motor function was assessed using a modified Bromage scale. Postoperative analgesia was standardised. Onset and duration of sensory and motor blockade, duration of analgesia, postoperative pain score, and analgesic requirement were compared. RESULTS: The clonidine patients showed an increase in duration of sensory loss from 489 min to 628 min with a mean difference of 138 min (95% confidence interval of 90 to 187 min), motor blockade from 552 min to 721 min with a mean difference of 170 min (95% confidence interval of 117 to 222 min), and analgesia from 587 min to 828 min with mean difference of 241 min (95% confidence interval of 188 to 294 min). There was no difference in onset time. No side effects were noted. CONCLUSION: The addition of 150 microg of clonidine to ropivacaine, for brachial plexus blockade, prolongs motor and sensory block and analgesia, without an increased incidence of side effects. PMID- 11032271 TI - Intraoperative loading attenuates nausea and vomiting of tramadol patient controlled analgesia. AB - PURPOSE: To evaluate the adverse effect profile of tramadol by patient-controlled analgesia (PCA) with administration of the loading dose either intraoperatively or postoperatively. METHODS: Sixty adult patients scheduled for elective abdominal surgery were enrolled into this prospective, randomized, double blind study. The patients were anesthetized in a similar manner. At the beginning of wound closure, the patients were randomly allocated to receive 5 mg x kg(-1) tramadol (Group 1) or normal saline (Group 2). In the post-anesthesia care unit (PACU), when patients in either group complained of pain, 30 mg x ml(-1) tramadol i.v. were given every three minutes until visual analogue scale (VAS) 3, followed by tramadol PCA with bolus dose of 30 mg and five minute lockout interval. Pain control and adverse effect assessments were done in the PACU and every six hours for 48 hr post drug by an independent observer. RESULTS: The loading dose was 290 +/- 45 mg in Group 1 and 315 +/- 148 mg in Group 2. In PACU, more nausea/vomiting both in terms of incidence (13/30, 43% vs 2/30, 6.6%, P < 0.05) and severity (nausea/vomiting score 2.5 +/- 2.0 vs 0.2 +/- 0.6, P < 0.05) was observed in patients with postoperative loading than in those with intraoperative loading of tramadol. CONCLUSION: Administering the loading dose of tramadol during surgery decreases the nausea/vomiting associated with high dose of tramadol and improves the quality of tramadol PCA in the relief of postoperative pain. PMID- 11032272 TI - Inadequate preanesthesia equipment checks in a simulator. AB - PURPOSE: To assess how completely anesthesiologists check their machinery and equipment before use, and to determine what influence seniority, age and type of practice may have on checking practices. METHODS: One hundred and twenty anesthesiologists were videotaped during a simulated anesthesia session. Each participant was scored by an assessor according to the number of items checked prior to the induction of anesthesia. A checklist of 20 items derived from well publicized, international standards was used. RESULTS: Participants were grouped according to their type of practice. Overall, mean scores were low. The ideal score was 20. There were no differences among university anesthesiologists (mean score 10.1, standard deviation 4.3), community anesthesiologists (7.5 +/- 4.3) and anesthesia residents (9.0 +/- 3.8). Each of these groups scored, on average, better than medical students (3.6 +/- 3.7) (P < 0.05). Neither age (r = 0.15, P > 0.1) nor number of years in practice (r = -0.18, P > 0.1) correlated with score. CONCLUSIONS: Our study suggests that the equipment-checking practices of anesthesiologists require considerable improvement when compared with national and international standards. Possible reasons for this are discussed and some remedial suggestions are made. PMID- 11032273 TI - "Deep-forehead" temperature correlates well with blood temperature. AB - PURPOSE: To evaluate the accuracy and precision of "deep-forehead" temperature with rectal, esophageal, and tympanic membrane temperatures, compared with blood temperature. METHODS: We studied 41 ASA physical status 1 or 2 patients undergoing abdominal and thoracic surgery scheduled to require at least three hours. "Deep-forehead" temperature was measured using a Coretemp thermometer (Terumo, Tokyo, Japan). Blood temperature was measured with a thermistor of a pulmonary artery. Rectal, tympanic membrane, and distal esophageal temperatures were measured with thermocouples. All temperatures were recorded at 20 min intervals after the induction of anesthesia. We considered blood temperature as the reference value. Temperatures at the other four sites were compared with blood temperature using correlation, regression, and Bland and Altman analyses. We determined accuracy (mean difference between reference and test temperatures) and precision (standard deviation of the difference) of 0.5 degrees C to be clinically acceptable. RESULTS: "Deep-forehead" temperature correlated well with blood temperature as well as other temperatures, the determination coefficients (r2) being 0.85 in each case. The bias for the "deep-forehead" temperature was 0.0 degrees C, which was the same as tympanic membrane temperature and was smaller than rectal and esophageal temperatures. The standard deviation of the differences for the "deep-forehead" temperature was 0.3 degrees C, which was the same as rectal temperature. CONCLUSIONS: We have demonstrated that the "deep forehead" temperature has excellent accuracy and clinically sufficient precision as well as other three core temperatures, compared with blood temperature. PMID- 11032274 TI - Occupational exposure to nitrous oxide and desflurane during ear-nose-throat surgery. AB - PURPOSE: To determine occupational exposure of the anesthesiologist and surgeon to nitrous oxide and desflurane during general anesthesia for ear-nose-throat (ENT) surgery in children and adults. METHODS: An observational clinical trial was performed in ten children (C) and ten adults (A). Tracheas were intubated, in adults, with cuffed tubes and in children with uncuffed tubes. The operating room was equipped with modern air conditioning and waste anesthetic gas scavengers. Gas samples were obtained during the operative procedure every 90 sec from the breathing zone of subjects. Time-weighted averages (TWA) over the time of exposure were calculated for nitrous oxide and desflurane. RESULTS: Nitrous oxide TWAs for anesthesiologists were 0.41 +/- 0.23 ppm (A) and 1.20 +/- 0.32 ppm (C, P < 0.0001), and 2.24 +/- 1.93 ppm (A) and 5.30 +/- 0.60 ppm (C, P = 0.0001) for the surgeon who worked close to the patient's airway and thus had higher exposure (P < 0.05 [A], P < 0.0001 [C]). With regard to desflurane, the anesthesiologists' TWAs were 0.02 +/- 0.03 ppm for both adults and children. The surgeon was exposed to 0.21 +/- 0.24 ppm desflurane (A) and 0.30 +/- 0.14 ppm (C, P: n.s.). Although the surgeon's exposure was greater (P < 0.05 [A], P < 0.0001 [C]), the threshold limits of 25 ppm for nitrous oxide and 2 ppm for desflurane recommended by the National Institute of Occupational Safety and Health were not exceeded. CONCLUSIONS: Under modern air conditioning, occupational exposure to inhalational anesthetics is low, and inhalational anesthesia is safe from the standpoint of modern workplace laws and health-care regulations. PMID- 11032275 TI - Intubating LMA guided awake fibreoptic intubation in severe maxillo-facial injury. AB - PURPOSE: To describe control of the airway in a patient with severe maxillo facial injury using intubating laryngeal mask guided awake fibreoptic intubation. CLINICAL FEATURES: A 110 kg man presented with a severe facial injury due to a horse kick. Local examination showed a large transverse gaping laceration just beneath the nose, exposing the lower nasal cavities through to the posterior nasopharynx. It was planned to perform a surgical tracheostomy before surgical fixation of the maxilla and nasal bones. Considering the patient's weight, short neck and difficulty in lying flat, surgical tracheostomy under general anesthesia was considered the best option. Under topical anesthesia, an intubating laryngeal mask was introduced followed by a fibreoptic bronchscope. The endotracheal tube was threaded over the bronchoscope and airway control achieved. CONCLUSIONS: The use of an intubating laryngeal mask avoids the need for manipulation of the head and neck and acts as a 'shield' for the fibreoptic bronchoscope from surrounding blood in patients with maxillo-facial injury. The use of the fibreoptic bronchoscope ensures intubation under vision. PMID- 11032276 TI - Reversal of paraparesis after thoracic aneurysm repair by cerebrospinal fluid drainage. AB - PURPOSE: To describe a case of reversal of delayed paraparesis, after an elective type I thoracoabdominal aortic aneurysm (TAAA) repair, via cerebrospinal fluid (CSF) drainage. CLINICAL FEATURES: A 75-yr-old woman developed paraparesis 13 hr after type I TAAA repair. The patient had been given combined regional and general anesthesia. There was no cerebrospinal fluid drain inserted at the time of surgery. The patient was hemodynamically stable throughout the procedure and was transported to the intensive care unit with trachea intubated and lungs ventilated. She demonstrated some initial lower limb paraparesis but had good recovery of limb function three hours after cessation of the epidural infusion. However, five hours and forty-five minutes after stopping the epidural, she was again paraparetic. Peripheral nerve injury, prolonged effects of epidural local anesthetic, and epidural hematoma were ruled out as precipitating factors. Cord ischemia was considered possible and a CSF catheter was inserted. Immediate improvement was seen upon catheter insertion and commencement of drainage, beginning with movement in the left toes and foot. Drainage was performed when the CFS pressure became > 15 mmHg. Motor function in the lower limbs continued to improve with each drainage extending to complete recovery after 40 hr. She was discharged home 11 days after surgery with no neurological deficit. CONCLUSION: Drainage of CSF was useful in treating a case of post-TAAA neurologic deficit. PMID- 11032277 TI - Intracerebral hemorrhage and Moyamoya disease in pregnancy. AB - PURPOSE: To present a case of Moyamoya disease with intracranial hemorrhage complicating pregnancy. CLINICAL FEATURES: A 36-yr-old parturient at 34 wk gestation presented with left hemiparesis, headache, nausea and vomiting. Subsequent deterioration in level of consciousness and the development of a dilated right pupil necessitated immediate intubation. Urgent non-contrast CT scan revealed a large right intracerebral hematoma with transtentorial herniation. The patient underwent simultaneous emergency Cesarean section and craniotomy. A postoperative angiogram revealed findings consistent with Moyamoya disease. The neonate survived but the patient developed severe cerebral edema and died eleven days postoperatively. CONCLUSION: Adult patients with Moyamoya disease often present with intracranial hemorrhage which poses unique anesthetic challenges. We report a case of intracerebral hemorrhage during pregnancy, which is known to be associated with high morbidity and mortality. The anesthetic techniques are reviewed and discussed. PMID- 11032278 TI - Leprosy and the anesthesiologist. AB - PURPOSE: To discuss particular aspects of leprosy (complications treatment, special population) that have implications for anesthetic management in leprous patients scheduled for surgery. SOURCE: MEDLINE and manual searches of relevant literature. Multiple MEDLINE searches (from 1966 onwards) were done, using LEPROSY as a common Medical Subject Heading (MeSH). Other headings used were: anesthesia, surgery, cardiovascular system, respiratory system, eye, skin, nervous system, genitalia, pathology, psychology and pregnancy. A large number of references were retrieved, but only 18 of them were relevant to our topic. Others were obtained by manual search and cross referencing. PRINCIPAL FINDINGS: Leprosy, especially lepromatous leprosy, is a systemic disease, affecting many organs and systems of the body, e.g., cardiovascular (cardiac dysautonomia), respiratory (impaired cough response, nasal obstruction), hepatobiliary (hepatitis), renal (nephritis), ocular (anesthesia), hematological (reduced red, white and platelet count) and osseous systems (bone resorption). CONCLUSION: Investigation of the systems likely to be affected by leprosy (e.g., complete hemogram, liver, lung and kidney function tests, Valsalva response, assessment of ocular anesthesia) should form part of a preanesthetic check up in patients with leprosy. PMID- 11032279 TI - 5-HT3 receptor antagonists vs traditional agents for the prophylaxis of postoperative nausea and vomiting. AB - PURPOSE: Numerous antiemetics have been studied for the prevention of postoperative nausea and vomiting (PONV) including traditional agents (metoclopramide, perphenazine, prochlorperazine, cyclizine and droperidol) and the 5-HT3 receptor antagonists (ondansetron, dolasetron, granisetron and tropisetron). The results have been divergent and inconsistent. The purpose of this quantitative systematic review was to evaluate the effectiveness of 5HT3 receptor antagonists compared to traditional antiemetics for the prevention of PONV METHODS: A systematic search of the English language literature using computerized MEDLINE, EMBASE, and Pre-MEDLINE databases from 1966 to October 1999 and a manual search of references from retrieved articles were performed. Individual efficacy and adverse effect data was extracted from each of the studies according to a predefined protocol. The summary odds ratios were calculated using the Dersimonian and Laird method under a random effects model. RESULTS: A total of 41 trials met our pre-defined inclusion criteria and were included in our analysis. Results in the 32 studies examining PONV indicated a 46% reduction in the odds of PONV in the 5-HT3-treated group (0.54 [95% CI 0.42 0.71], P < 0.001). Evaluation of PONV by traditional antiemetic agent demonstrated a 39% reduction compared with droperidol (0.61 [95% CI 0.42-0.89], P < 0.001) and a 56% reduction compared with metoclopramide (0.44 [95% CI 0.31 0.62], P < 0.001). Results in the 34 studies examining vomiting indicated a 38% reduction in the odds of vomiting in the 5-HT3-treated group (0.62 [95% CI 0.48 0.81], P < 0.001). CONCLUSIONS: The 5-HT3 receptor antagonists are superior to traditional antiemetic agents for the prevention of PONV and vomiting. The reduction in the odds of PONV and vomiting is significant in the overall analysis and the subgroup analyses comparing 5-HT3 receptor antagonists with droperidol and metoclopramide. PMID- 11032280 TI - Ketamine suppresses endotoxin-induced NF-kappaB expression. AB - PURPOSE: Ketamine reduces endotoxin-induced production of proinflammatory cytokines, including tumour necrosis factor-alpha (TNF), in several types of inflammatory cells, including monocytes and macrophages. Transcription of the genes that encode production of these proinflammatory cytokines is regulated by nuclear factor-kappa B (NF-kappaB). Cytoplasmic B protein is activated by endotoxin (LPS) as well as by TNF, allowing B protein to migrate into the cell nucleus to activate gene transcription for these inflammatory mediators. Because NF-kappaB is likely involved in brain injury and inflammatory neurodegenerative disease, such as multiple sclerosis, we examined whether ketamine inhibits LPS induced activation of NF-kappaB in human glioma cells in vitro and intact mouse brain cells in vivo. METHODS: Endotoxin-induced NF-kappaB expression in both the human glioma cells in vitro and the intact mouse brain cells in vivo was determined by electrophoretic mobility shift assays (EMSA) of nuclear extracts and measurement of NF-kappaB expression by densitometry. Endotoxin was injected intracerebroventricularly in vivo and intact brain was harvested. Klenow fragment labeling was used to identify NF-kappaB protein for both the in vivo and vitro experiments. RESULTS: Endotoxin treatment increased NF-kappaB expression (P < 0.05) both in vivo and vitro compared with control (untreated) cells. Ketamine suppressed endotoxin-induced neuronal NF-kappaB activation in a dose-dependent manner (P < 0.05, except for the 10(-5) M concentration in vitro) both in vivo and vitro. CONCLUSION: Ketamine inhibits endotoxin-induced NF-kappaB expression in brain cells in vivo and vitro and it is suggested that this may have implications in the neuroprotective effects of ketamine reported by other investigators. PMID- 11032281 TI - Propofol prevents lipid peroxidation following transient forebrain ischemia in gerbils. AB - PURPOSE: To ascertain whether propofol prevents lipid peroxidation on delayed neuronal death induced by transient forebrain ischemia in the hippocampal CA1 subfield in gerbils. METHODS: Forty gerbils were randomly assigned to five groups: Group I, control, sham operation treated with physiological saline solution (PSS); Group II, ischemia/reperfusion treated with PSS; Group III, ischemia/reperfusion treated with 50 mg x kg(-1) propofol; Group IV, ischemia/reperfusion treated with 100 mg x kg(-1) propofol; Group V, ischemia/reperfusion treated with 150 mg x kg(-1) propofol. Transient forebrain ischemia was induced by occluding the bilateral common carotid arteries for four minutes under N2O/O2/halothane anesthesia after propofol or PSS. Five days later, the cerebrum was removed and each forebrain was cut into two including the hippocampus. Lipid peroxidation was determined using the production of malondialdehyde (MDA), and histopathological changes in the hippocampal CA1 subfield were examined. RESULTS: In group II, the pyramidal cells were atrophic and pycnotic; vacuolation and structural disruption of the radial striated zone was observed. In the other four groups, these changes were not observed. Degenerative ratios of pyramidal cells were: Group I: 4.9 +/- 2.3, Group II: 94.1 +/- 4.5 (P < 0.01), Group III: 12.5 +/- 5.7, Group IV: 11.0 +/- 4.6, Group V: 9.6 +/- 4.9%. Production of MDA was: Group I: 83 +/- 22, Group II: 198 +/- 25 (P < 0.01), Group III: 153 +/- 39, Group IV: 113 +/- 34, Group V: 106 +/- 27 nmol x g( 1) wet tissue. CONCLUSION: Propofol attenuated delayed neuronal death by preventing lipid peroxidation induced by transient forebrain ischemia in the hippocampal CA1 subfield in gerbils. PMID- 11032282 TI - Syringe size and flow rate affect drug delivery from syringe pumps. AB - PURPOSE: To evaluate the effect of syringe size and infusion rate on drug delivery after vertical displacement of syringe pumps. METHODS: Four syringes (10 ml, 20 ml, 30 ml, 50 ml) were studied at three infusion speeds (2 ml x hr(-1), 1 ml x hr(-1) and 0.5 ml x hr(-1)). Fluid delivery was measured gravimetrically using an electronic balance. Aspiration volume (amount of fluid retracted into the syringe-infusion line assembly) and zero-drug delivery time (time between the lowering of the syringe pump and reattainment of the initial weight) were determined after lowering the syringe pump 130 cm or 50 cm. Then, infusion bolus was measured after elevating the pump to its initial position. Syringe compliance was calculated from the occlusion release bolus at an occlusion pressure of 300 mmHg. RESULTS: Zero-drug delivery time, aspiration volume and infusion bolus increased with syringe size (P < 0.0001). At 0.5 ml x hr(-1), a zero-drug delivery time of 29.7 +/- 0.6 min was recorded for the 50 ml syringe and 5.5 +/- 0.15 min for the 10 ml syringe. As infusion rates increased, zero-drug delivery times decreased following a 1/x ratio. Zero-drug delivery time highly correlated with calculated syringe compliance (Pearson's r = 0.97, P < 0.001), which was closely related to syringe plunger area (R2 = 0.93, P < 0.001). CONCLUSIONS: Vertical displacement of syringe pumps causes flow irregularities which correlate directly with syringe size and indirectly with infusion rate. Extreme vertical maneuvering of syringe pumps should be avoided. Infusion rates > or = 1 ml x hr( 1) and low-compliant, small syringes are recommended when highly concentrated drugs are administered. PMID- 11032283 TI - Laureates of the history of anesthesia, 2000. PMID- 11032284 TI - Canadian Anesthesiologists' Society Gold Medal. Dr. Tom McCaughey. PMID- 11032286 TI - Computerized EEG monitoring of anesthetic depth: Quo Vadis? PMID- 11032285 TI - Canadian Anesthesiologists' Society Gold Medal. Dr. Graham McMorland. PMID- 11032287 TI - Herbals--beware bleeding. PMID- 11032288 TI - Unusual cause of volotrauma. PMID- 11032289 TI - Thin end of the wedge. PMID- 11032290 TI - An unusual cause of difficult spinal anesthesia. PMID- 11032291 TI - Tinnitus and insomnia. AB - PURPOSE: To investigate the effects of insomnia on tinnitus severity and to determine how this relationship may evolve with the passage of time. MATERIALS AND METHODS: Questionnaires were mailed to patients before their initial appointment at the Oregon Health Sciences University Tinnitus Clinic between 1994 and 1997. These questionnaires requested information pertaining to insomnia, tinnitus severity, and loudness. During their initial appointment, patients received counseling, education, and reassurance about tinnitus; audiometric and tinnitus evaluations; and treatment recommendations. Follow-up questionnaires were mailed to 350 patients 1 to 4 years (mean = 2.3 yr) after their initial appointment at the clinic. RESULTS: One hundred seventy-four patients (130 men, 44 women; mean age 55.9 yr) returned follow-up questionnaires. Although many of these patients improved in both sleep interference and tinnitus severity, a significant number (43) reported on the follow-up questionnaire that they continued to have difficulty sleeping. Reported loudness and severity of tinnitus were significantly greater for this group than for groups of patients who reported that they never or only sometimes have difficulty sleeping. The relationship between sleep disturbance and tinnitus severity became more pronounced with the passage of time. CONCLUSIONS: Insomnia is associated with greater perceived loudness and severity of tinnitus. These findings underscore the importance of identification and successful treatment of insomnia for patients with tinnitus. PMID- 11032292 TI - Anterior commissure carcinoma: I-histopathologic study. AB - OBJECTIVES: To study the behavior of anterior commissure carcinoma regarding its tendency for cartilage invasion. METHODS: Histopathological examination of the region of the anterior commissure (AC) was done with whole organ section in 30 randomly selected specimens. Serial sections were prepared and examined histopathologically for evidence of microscopic cartilage invasion in the region of the AC. RESULTS: Microscopic involvement of the thyroid cartilage was detected in the 30 sections studied. CONCLUSION: Whether recurrence was de novo or initiated by residual malignant cells, it is mandatory to excise the anterior portion of the thyroid cartilage with the tumor-bearing mucosa. PMID- 11032293 TI - Computed tomography vs. magnetic resonance imaging of acute bacterial sinusitis: a rabbit model. AB - PURPOSE: Computed topography (CT) and magnetic resonance imaging (MRI) are important, both clinically and in a research setting, in assessing bacterial sinusitis (BS). The use of CT scanning to evaluate sinus opacification in a reversible model of rabbit acute sinusitis has been reported. MRI offers the potential for better visualization of soft tissue and fluid changes within the paranasal sinuses. MRI has potential as a research tool in animal models of sinusitis. This article compares the use of CT and MRI in measuring maxillary sinus opacification in rabbits during experimental, reversible BS. MATERIALS AND METHODS: In 2 independent trials, New Zealand White rabbits were imaged for baseline anatomy, and BS was generated by sinus inoculation with Staphylococcus aureus. Serial imaging was performed as a measure of the progression and resolution of BS during the trials. Two experienced, independent reviewers then scored each CT and MRI for percent opacification of the maxillary sinus. These scores were analyzed to assess the degree of agreement between the reviewers. RESULTS: The correlation coefficients for CT and MRI were 0.6816 and 0.3584, respectively. The Z-statistic comparing these correlation coefficients was significant (P < .0001), indicating that CT is a more precise measure of reversible BS in this rabbit model. Differences in mean scan time and cost per scan were also significantly different (P < .0001), with CT being both quicker and less expensive. CONCLUSIONS: Greater interobserver consistency of scan interpretation, with less time and cost, make CT the preferred tool for measuring BS in this rabbit model. Attributes of MRI such as better resolution of fluid tissue interfaces and custom surface coil design for visualization of specific anatomic structures are discussed as they may increase the effectiveness of MRI as an imaging modality in future sinusitis research. PMID- 11032294 TI - Concurrent chemotherapy and "concomitant boost" radiotherapy for unresectable head and neck cancer. AB - PURPOSE: For patients with advanced head and neck cancer, various combined chemoradiotherapy regimens have been used to improve local control. This study was carried out to assess the outcome of concomitant chemotherapy with a "concomitant boost" radiotherapy in the treatment of advanced unresectable head and neck cancer patients. MATERIALS AND METHODS: Forty-eight patients were treated with combined chemoradiotherapy between the years of 1990 and 1995. Cisplatinum (100 mg/m2) was given intravenously during week 1 and week 5. A "concomitant boost" external beam radiotherapy approach was used with twice-daily treatment delivered during the last 2 weeks. A total of 70 Gy was delivered over 6 weeks. Median follow-up was 23.5 months (2-79 months). RESULTS: Thirty-one (65%) and 17 (35%) patients achieved complete and partial response, respectively. Median survival in complete responders has not been reached. Overall survival at 2 years, 3 years, and 5 years were 58.7%, 52.8%, and 42.4%, respectively. Median overall survival was 38.8 months. Acute confluent mucositis (Radiation Therapy Oncology [RTOG] grade 3) developed in 50% of patients, but there was no severe long-term treatment-related toxicity. CONCLUSION: This combined chemoradiotherapy approach is safe and efficacious for advanced unresectable head and neck cancer. Treatment-related toxicity was acceptable with 50% of patients developing acute confluent mucositis. There was no severe long-term treatment-related toxicity. PMID- 11032295 TI - Morbidity after midline mandibulotomy and radiation therapy. AB - PURPOSE: To assess the morbidity of mandibulotomy in patients treated for neoplasms of the oropharynx and oral cavity, and to determine if postoperative radiation therapy to the mandibulotomy site carries an increased risk of complications. PATIENTS AND METHODS: The medical charts of 30 patients treated between 1992 and 1996 undergoing midline mandibulotomy for tumors of the oral cavity (7 patients) and oropharynx (23 patients) were retrospectively reviewed. Three patients presented with recurrent disease, 1 of whom was previously irradiated. Twenty-five patients received postoperative radiation after mandibulotomy to a median dose of 60 Gy to the primary tumor bed, whereas 5 patients were treated with surgery alone. The patients were separated into those whose mandibulotomy site was within the radiation treatment field (n = 9), and those whose site was shielded (n = 10). Median follow-up was 27.8 months (range 5 81 months). End points included significant pain involving the mandibulotomy site, trismus, malocclusion, wound infection, osteoradionecrosis, and time to oral intake. RESULTS: There were no postoperative deaths. Minor wound infection or breakdown occurred in 4/30 patients (13%). All of these resolved with local care and parenteral antibiotics. More serious complications involving the mandibulotomy occurred in 2 patients (7%). One patient had chronic wound drainage at the mandibular osteotomy site, which healed after plate removal. Another patient developed osteoradionecrosis. No patient developed trismus or malocclusion. With a median follow-up of 27.8 months, 4 patients have recurred locally. The complication rate was 11% for patients whose mandibulotomy site was irradiated, and 30% for those whose site was shielded. CONCLUSION: Mandibulotomy can be safely performed in patients who are likely to require postoperative external radiation. PMID- 11032296 TI - The omohyoid sling syndrome. AB - BACKGROUND: Patients with the omohyoid syndrome present with a transient lower neck swelling during swallowing. The mechanism of functional derangement has been hypothesized previously but never proven. Our aim is to show how this can be demonstrated real-time. METHOD AND PATIENTS: Five patients were studied. Serial photography, clinical videography, real-time ultrasonography, and simulated cine mode computed tomography were used. Findings were compared with the normal side. The literature was reviewed. RESULTS: Very little literature experience is available. The swelling appears when the throat ascends and subsides with its descent in unison with omohyoid prominence. The sternomastoid is passively tented up by an abnormal underlying omohyoid. This latter appears to have lost its restriction to bowstring by the retaining deep cervical fascia. CONCLUSION: The described approach allowed for definite proof of the pathogenetic mechanism, provided a means of diagnosis, and suggested further insight into the management of these patients. PMID- 11032297 TI - A new role for computed tomography in the diagnosis and treatment of pyriform sinus fistula. AB - Neck abscess located in or around the thyroid gland should raise the suspicion of acute suppurative thyroiditis, pyriform sinus fistula, a 3rd or 4th branchial cleft anomaly. Differentiating between these entities on a clinical basis is difficult. After treating the initial infection, computed tomography, barium esophagography, ultrasound, and endoscopy can be used in search of the fistulous tract that can be associated with a pyriform sinus fistula or a 3rd or 4th branchial anomaly. We present a case of a pyriform sinus fistula involving the use of oral contrast, combined with computed tomography, to delineate the tract and its surrounding structures. This method, in combination with endoscopy, aided in the removal of this branchial anomaly. PMID- 11032298 TI - Structural airway anomalies in patients with DiGeorge syndrome: a current review. AB - DiGeorge Syndrome is a genetic disorder characterized by either absence or hypoplasia of the thymus and the parathyroid glands. Patients with this syndrome also have a high incidence of cardiovascular malformations and facial dysmorphism. Structural airway anomalies have also been described, albeit infrequently. Tracheoesophageal fistula, short trachea with reduced numbers of tracheal rings, abnormal thyroid cartilage, laryngomalacia, tracheomalacia, and bronchomalacia have been recognized in these patients. We review all previously reported patients with DiGeorge syndrome and lower airway anomalies. In addition, we present 2 patients with DiGeorge syndrome who were each found to have an aberrant right tracheal bronchus. Structural airway anomalies can be a cause of morbidity and mortality in patients with DiGeorge syndrome. Prompt, thorough evaluation of the upper and lower airway in these patients is essential. PMID- 11032299 TI - Temporomandibular joint cyst as a preauricular mass. PMID- 11032300 TI - Tracheoesophageal fistula secondary to disc battery ingestion. PMID- 11032301 TI - Spontaneous subcutaneous temporal emphysema. AB - The presence of air in the temporal regions is an uncommon presentation of spontaneous pneumomediastinum. As terminal alveoli rupture, air dissects along bronchovascular shafts. Usually, air travels either in a superior or inferior direction. Thus, extensive spontaneous subcutaneous emphysema accompanied by both cervical and retroperitoneal emphysema is rarely encountered. We present an unusual case of spontaneous pneumomediastinum, pneumoretroperitoneum, and cervical and facial emphysema presenting as bilateral painless temporal swelling. To our knowledge, this association has not been reported. Treatment involves observation for potentially life-threatening sequelae. PMID- 11032302 TI - Earpick injury of the stapes. PMID- 11032303 TI - Management of impaired vocal fold movement during sleep in a patient with Shy Drager syndrome. AB - A 46-year-old woman with Shy-Drager syndrome is presented. She has impaired vocal fold abduction during sleep, but has no laryngeal dysfunction while she is awake. In order to reduce laryngeal obstruction during sleep, she initially underwent laterofixation of 1 vocal fold (Ejnell's method) with little lasting success because of accidental slipping of the ligature. Later, she successively underwent arytenoidectomy with the use of CO2 laser. Her noctural breathing improved markedly after arytenoidectomy. PMID- 11032304 TI - Potpourri aspiration presenting as tension pneumothorax. AB - Foreign body aspiration in children is a relatively common occurrence, with peanuts, seeds, or other food particles representing the most common items. Because radiological findings such as mediastinal shift, postobstructive emphysema, and pneumonia are notoriously inconsistent, diagnosis hinges on an accurate history, which may be correlated by physical examination and radiography. We present the case of a 2-year-old girl with delayed treatment of a bronchial foreign body who presented with tension pneumothorax before endoscopy. After chest tube removal, her pneumothorax recurred, thereby bringing about the question of bronchial erosion. Furthermore, an uncommonly reported aspirated object, household potpourri, was encountered. PMID- 11032305 TI - DNA methylation in ciliates: implications in differentiation processes. AB - Much experimental evidence on the role of DNA methylation in gene expression has been reported. Here we review reports on DNA methylation in ciliated protozoa, emphasizing its implications in cell differentiation processes. Both types of methylated bases (adenine and cytosine) can be found in macronuclear DNA. The division cycle and conjugation have been studied with regard to adenine methylation, and several different functions have been assigned to the methylation changes detected in these processes. Cytosine methylation changes were analyzed during stomatogenesis of Paramecium and encystment of Colpoda inflata. A comparative analysis with other similar microbial eukaryotic differentiation processes is carried out. PMID- 11032306 TI - Molecular tools for breeding basidiomycetes. AB - The industrial production of edible basidiomycetes is increasing every year as a response to the increasing public demand of them because of their nutritional properties. About a dozen of fungal species can be currently produced for food with sound industrial and economic bases. Notwithstanding, this production is threatened by biotic and abiotic factors that make it necessary to improve the fungal strains currently used in industry. Breeding of edible basidiomycetes, however, has been mainly empirical and slow since the genetic tools useful in the selection of the new genetic material to be introduced in the commercial strains have not been developed for these fungi as it was for other organisms. In this review we will discuss the main genetic factors that should be considered to develop breeding approaches and tools for higher basidiomycetes. These factors are (i) the genetic system controlling fungal mating; (ii) the genomic structure and organisation of these fungi; and (iii) the identification of genes involved in the control of quantitative traits. We will discuss the weight of these factors using the oyster mushroom Pleurotus ostreatus as a model organism for most of the edible fungi cultivated industrially. PMID- 11032307 TI - Streptomycetes: a new model to study cell death. AB - Colonies of streptomycetes are now viewed as multicellular entities containing morphologically and biochemically differentiated cell types which have specific functions and precise spatial relationships to one another. Like multicellular organisms, colony development in streptomycetes is also maintained by a tight balance between cell proliferation and cell death processes. This review describes the current state of knowledge concerning cell death in streptomycetes. PMID- 11032308 TI - Pulsed-field gel electrophoresis analysis of virus assemblages present in a hypersaline environment. AB - A method for analyzing virus assemblages in aquatic environments was developed and used for studying the highest-salinity ponds (from 13.4 to 35% salinity) from a multi-pond solar saltern in Alicante, Spain. The protocol consisted of a series of concentration and purification steps including tangential flow filtration and ultracentrifugation, followed by the preparation of total viral nucleic acids that were subsequently separated by pulsed-field gel electrophoresis. For every sample analyzed, a characteristic DNA pattern was obtained, whose complexity was related to viral diversity. The comparison of our results with a similar analysis carried out with marine virus assemblages shows that, as expected, the viral diversity corresponding to the analyzed hypersaline environment is considerably lower than that of a marine environment. PMID- 11032309 TI - Phylogenetic position of parabasalid symbionts from the termite Calotermes flavicollis based on small subunit rRNA sequences. AB - Small subunit rDNA genes were amplified by polymerase chain reaction using specific primers from mixed-population DNA obtained from the whole hindgut of the termite Calotermes flavicollis. Comparative sequence analysis of the clones revealed two kinds of sequences that were both from parabasalid symbionts. In a molecular tree inferred by distance, parsimony and likelihood methods, and including 27 parabasalid sequences retrieved from the data bases, the sequences of the group II (clones Cf5 and Cf6) were closely related to the Devescovinidae/Calonymphidae species and thus were assigned to the Devescovinidae Foaina. The sequence of the group I (clone Cf1) emerged within the Trichomonadinae and strongly clustered with Tetratrichomonas gallinarum. On the basis of morphological data, the Monocercomonadidae Hexamastix termitis might be the most likely origin of this sequence. PMID- 11032310 TI - Effect of the fungicide benomyl on spore germination and hyphal length of the arbuscular mycorrhizal fungus Glomus mosseae. AB - The fungicide benomyl inhibited spore germination and hyphal length of the arbuscular mycorrhizal fungus Glomus mosseae when applied at doses of 21.25 microg/ml (agronomic dose), 10.62 microg/ml and 10 microg/ml. G. mosseae was able to germinate in the presence of 2.12 microg/ml of benomyl, and the percentage of spore germination was unaffected by dosis of 0.1, 0.01 and 0.001 microg/ml of the fungicide. However, all doses of fungicide tested in this study decreased the hyphal length. When ungerminated G. mosseae spores previously exposed to benomyl were transferred to water-agar medium without benomyl, the maximum germination was 16%. Small spores of G. mosseae were more resistant to benomyl than the larger ones. Our results show some of the factors which can explain the variability of the effect of benomyl on arbuscular mycorrhizal fungi. PMID- 11032311 TI - Binding of soluble glycoproteins from sugarcane juice to cells of Acetobacter diazotrophicus. AB - Sugarcane produces two different pools of glycoproteins containing a heterofructan as glycidic moiety, tentatively defined as high-molecular mass (HMMG) and mid-molecular mass (MMMG) glycoproteins. Both kinds of glycoproteins can be recovered in sugarcane juice. Fluorescein-labelled glycoproteins are able to bind to Acetobacter diazotrophicus cells, a natural endophyte of sugarcane. This property implies the aggregation of bacterial cells in liquid culture after addition of HMMG or MMMG. Anionic glycoproteins seem to be responsible for the binding activity whereas cationic fraction is not retained on the surface ofA. diazotrophicus. Bound HMMG is competitively desorbed by sucrose whereas MMMG is desorbed by glucosamine or fructose. On this basis, a hypothesis about the discriminatory ability of sugarcane to choose the compatible endophyte from several possible ones is proposed. PMID- 11032312 TI - Colorimetric in situ hybridization (CISH) with digoxigenin-labeled oligonucleotide probes in autofluorescent hyphomycetes. AB - We used digoxigenin-labeled probes for in situ hybridization of hyphomycetes to replace the commonly used fluorescent proof of probe binding by a colorimetric reaction. The resulting blue-purple, intracellular precipitate could be easily detected by light microscopy, and thus presented a promising method to overcome autofluorescence of fungal material and substratum. Optimal cell fixation and permeabilization procedures, as well as hybridization conditions were developed on the example of two different hyphomycetes: Phialophora sp. and Cartapip, a colorless mutant of Ophiostoma piliferum (Agra Sol). PMID- 11032313 TI - Sulfite-reducing clostridia in the sediment of a high mountain lake (Laguna Grande, Gredos, Spain) as indicators of fecal pollution. AB - We studied the vertical distribution of sulfite-reducing clostridia in the sediment of a Spanish high-mountain lagoon (Laguna Grande de Gredos, central Spain), with optimal sediment characteristics (temperature < 20 degrees C) to maintain spores without growing. This allowed us to assess the original numbers of sulfite-reducing clostridia endospores settled, without postdepositional growing. Sulfite-reducing clostridia are normal inhabitants of the intestinal microbiota of humans and other mammals. These microorganisms may form endospores, which allow the bacteria to survive in almost any habitat, either terrestrial or aquatic, waiting for favorable conditions for growth. Sulfite-reducing clostridia could be suitable indicators of past human pollution because they have a great longevity in natural habitats and they cannot multiply at temperatures below 20 degrees C or in the presence of O2. We found a great increase in the numbers of clostridia (expressed as colony-forming units per gram [CFU/g] of dry weight of sediment) since the 1970s, which reflects the rise of human pressure caused by the practice of outdoor activities. Clostridia CFU/g rose dramatically after the faulty operation of the depuration system of a mountain refuge built close to the lagoon. We compared the vertical distribution of clostridia CFU/g from Laguna Grande sediments with those from a neighbor lagoon (Laguna Cimera), which showed less tourist pressure and no direct disposal of sewage. Finally, we agree with the usefulness of the numbers of sulfite-reducing clostridia as indicators of past pollution. PMID- 11032314 TI - The corollary of scientific research: "Publish or be damned". PMID- 11032315 TI - Effective medium approximation and deposition of colloidal particles in fibrous and granular media AB - Laminar flow of fluids through fibrous and granular media and deposition of colloidal particles from a liquid suspension are two fundamental phenomena encountered in many industrial applications. An Effective Medium Approximation (EMA) is used to determine the fluid flow permeability and particle capture efficiency of random arrays of cylindrical and spherical collectors. The EMA assumes a model system in which a packing element (a single fiber in the fibrous medium and a single sphere in the granular medium) is surrounded by a fluid envelope and an effective-medium beyond the envelope. It integrates the important features of both the cell models and Brinkman's model. The Stokes equation and Brinkman equation are solved for the fluid envelope and effective medium regions, respectively, to obtain the permeability and close-to-surface velocity field around the collectors. The convective diffusion equation is then solved to determine the particle deposition rate. The analytical expressions for the permeability and particle deposition rate are derived for all possible cases of random packing of uniform and non-uniform cylinders and spheres. Effects of various system properties and operating conditions on deposition of colloidal particles are investigated. The physical or chemical conditions include the properties which affect the magnitude of double layer interaction: the electrolyte concentration and surface potentials, and the property which affects the van der Waals interaction: the Hamaker constant. It was found that the effects of the above properties is much more significant when the surface interactions play more important roles in the particle deposition process, or when the height of the total interaction energy barrier is higher than 5 kBT. Particle deposition becomes virtually impossible when the height of the repulsive energy barrier increases beyond 20 kBT. PMID- 11032316 TI - Sex chromosome differentiation revealed by genomic in-situ hybridization. AB - In this work, genomic in-situ hybridization (GISH) was used to study the sex chromosome molecular differentiation on chromosomes of male and female individuals of the isopod crustacean Asellus aquaticus. As a composite hybridization probe, we contemporaneously used male and female whole genomic DNA differently labelled in the presence of an excess of unlabelled DNA of the female homogametic sex. The karyotype of A. aquaticius normally displays eight homomorphic chromosome pairs, but a heteromorphic sex chromosome pair is present in about a quarter of the males of a natural population previously identified by us. GISH did not reveal any sex chromosome molecular differentiation on the male and female homomorphic sex chromosome pair, and the karyotypes of these individuals were equally labelled by the male- and female-derived probe, while the heteromorphic Y chromosome showed a differentially labelled region only with the male-derived probe. This region evidently contains male-specific sequences but, because no similar hybridized region is observed on the male homomorphic chromosome pair, they are probably not important for sex determination but represent a molecular differentiation acquired from the Y chromosome. PMID- 11032317 TI - Molecular cloning and characterization of the Fugu rubripes MEST/COPG2 imprinting cluster and chromosomal localization in Fugu and Tetraodon nigroviridis. AB - We isolated Fugu genomic clones using the human MEST (Mesoderm-Specific Transcript) cDNA as probe. Sequence analysis revealed the presence of MEST and three additional genes which show homology to plant DNBP (DNA-Binding Protein), vertebrate COPG2 (Coat Protein Gamma 2), as well as to human and mouse UCN (Urocortin). Structures of Fugu and human MEST, COPG2 and UCN genes are very similar. Since MEST and COPG2 are neighboring genes on human chromosome 7q32, we can conclude that we identified their orthologs and that linkage of these genes is evolutionarily conserved in vertebrates. Unlike human MEST which underlies isoform-specific imprinting and is methylated in a parent-of-origin-specific fashion, the CpG island of the Fugu ortholog is completely methylated. The translation start of Fugu MEST is identical to the non-imprinted human isoform which is in good agreement with the assumption that genomic imprinting is restricted to mammals. Comparative mapping of these genes by fluorescence in-situ hybridization to metaphase chromosomes of Fugu rubripes and Tetraodon nigroviridis showed clear signals on one of the smallest acrocentric chromosomal pairs, which in Fugu, can be easily classified by its unique triangular shape. PMID- 11032318 TI - Structure and evolution of supernumerary chromosomes in the Pacific giant salamander, Dicamptodon tenebrosus. AB - We examined the genetic make-up and plausible origins of the supernumerary (B) chromosomes of the Pacific giant salamander, Dicamptodon tenebrosus, from the Pacific Northwest of North America. These salamanders have variable numbers of B chromosomes, from 0 to 10 per individual. Salamanders from the most southerly and northerly regions of the species' range have lower average numbers of B chromosomes than salamanders in the middle of the range. To assess how the supernumerary chromosomes originated in D. tenebrosus, B chromosome DNA was isolated by microdissection and amplified by degenerate oligonucleotide-primed PCR. The B chromosome DNA hybridized similarly to genomic DNA from individuals of D. tenebrosus and the related species D. copei and D. ensatus, thus demonstrating that the supernumerary chromosomes were derived from the normal chromosome complement. Unique hybridization bands in both D. copei and D. tenebrosus suggest that the shared sequences have evolved independently. PMID- 11032319 TI - Spatial arrangement of genes, centromeres and chromosomes in human blood cell nuclei and its changes during the cell cycle, differentiation and after irradiation. AB - Higher-order compartments of nuclear chromatin have been defined according to the replication timing, transcriptional activity, and information content (Ferreira et al. 1997, Sadoni et al. 1999). The results presented in this work contribute to this model of nuclear organization. Using different human blood cells, nuclear positioning of genes, centromeres, and whole chromosomes was investigated. Genes are located mostly in the interior of cell nuclei; centromeres are located near the nuclear periphery in agreement with the definition of the higher-order compartments. Genetic loci are found in specific subregions of cell nuclei which form distinct layers at defined centre-of-nucleus to locus distances. Inside these layers, the genetic loci are distributed randomly. Some chromosomes are polarized with genes located in the inner parts of the nucleus and centromere located on the nuclear periphery; polar organization was not found for some other chromosomes. The internal structure of the higher-order compartments as well as the polar and non-polar organization of chromosomes are basically conserved in different cell types and at various stages of the cell cycle. Some features of the nuclear structure are conserved even in differentiated cells and during cellular repair after irradiation, although shifted positioning of genetic loci was systematically observed during these processes. PMID- 11032320 TI - Gametocidal factor-induced structural rearrangements in rye chromosomes added to common wheat. AB - The gametocidal factor on the Aegilops cylindrica chromosome 2Cc was used to induce and analyze the nature of chromosomal rearrangements in rye chromosomes added to wheat. For this purpose we isolated plants disomic for a given rye chromosome and monosomic for 2Cc and analyzed their progenies cytologically. Rearranged rye chromosomes were identified in 7% of the progenies and consisted of rye deficiencies (4.6%), wheat rye dicentric and rye ring chromosomes (1.8%), and terminal translocations (0.6%). The dicentric and ring chromosomes initiated breakage-fusion-bridge cycles (BFB) that ceased within a few weeks after germination as the result of chromosome healing. Of 56 rye deficiencies identified, after backcrossing and selfing, only 33 were recovered in either homozygous or heterozygous condition covering all rye chromosomes except 7R. The low recovery rate is probably caused by the presence of multiple rearrangements induced in the wheat genome that resulted in poor plant vigor and seed set, low transmission, and an underestimation of the frequency of wheat rye dicentric chromosomes. Genomic in-situ hybridization (GISH) analysis of the 33 recovered rye deficiencies revealed that 30 resulted from a single break in one chromosome arm followed by the loss of the segment distal to the breakpoint. Only three had a wheat segment attached distal to the breakpoint. Although some of the Gc induced rye rearrangements were derived from BFB cycles, all of the recovered rye rearrangements were simple in structure. The healing of the broken chromosome ends was achieved either by the de-novo addition of telomeric repeats leading to deficiencies and telocentric chromosomes or by the fusion with other broken ends in the form of stable monocentric terminal translocation chromosomes. PMID- 11032321 TI - Common methylation characteristics of sex chromosomes in somatic and germ cells from mouse, lemur and human. AB - DNA methylation of sex chromosomes was analysed using anti-5-methylcytosine antibodies on metaphase chromosomes of somatic cells from three species: human, lemur and mouse. Germ cells were also studied in male mouse. In female cells (human and mouse), the late replicating X was always the less methylated chromosome. Compared with autosomes, the methylation of both X chromosomes was always lower in fibroblasts than in lymphocytes and the difference was always greater in mouse than in human. In human, mouse and lemur male cells, the labelling of the unique X chromosome was quite similar to that of the early replicating X from female cells. Except for the heterochromatic region of the human Y chromosome, strongly methylated, the overall methylation of the Y chromosome was low. In mouse testicular cells, a variety of DNA methylation patterns was observed according to the cell type and the state of differentiation. Finally, the only structures of sex chromosomes which remain methylated in all conditions correspond to their pseudoautosomal regions. PMID- 11032322 TI - Comparative analysis of DNA methylation in tobacco heterochromatic sequences. AB - Cytosine methylation levels and susceptibility to drug-induced hypomethylation have been studied in several Nicotiana tabacum (tobacco) DNA repetitive sequences. It has been shown using HapII, MspI, BamHI and Sau3AI methylation sensitive restriction enzymes that the degree of 5'-mCmCG-3' methylation varied significantly between different repeats. There were almost saturation levels of 5 methylcytosine at the inner (3') cytosine position and variable degrees of methylation at the outer (5') cytosine at the enzyme recognition sites. The non transcribed high copy satellite sequences (HRS60, GRS) displayed significant heterogeneity in methylation of their basic units while middle repetitive sequences (R8.1, GRD5, 5S rDNA) were more uniformly modified at both cytosine residues. Dihydroxypropyladenine (DHPA) treatment, which is thought to reduce DNA methyltransferase activity by increasing S-adenosylhomocysteine levels, resulted in extensive demethylation of the outer cytosine in all repeats, and the partial hypomethylation of cytosines at the inner positions in less densely methylated repeats such as HRS60 and GRS. The results suggest that hypomethylation of 5' mCmCG-3' sites with DHPA is a gradual non-random process proceeding in the direction mCmCG-->CmCG-->CCG. The 18S-5.8S-25S rDNA was remarkably hypomethylated relative to the 5S rDNA at all restriction sites studied. Fluorescence in-situ hybridization showed that DNA decondensation within and between the 18S-5.8S-25S and 5S rDNA loci was variable in different nuclei. All nuclei had condensed and decondensed sequence. The chromatin of 18S-5.8S-25S rDNA was more readily digested with micrococcal nuclease than the 5S rDNA suggesting that the overall levels of decondensation were higher for 18S-5.8S-25S rDNA. Variable decondensation patterns within and between loci were also observed for GRS and HRS60. Cytosine methylation of the tobacco repeats is discussed with respect to transcription, overall levels of condensation and overall structure. PMID- 11032323 TI - Neocentric activity of rye 5RL chromosome in wheat. AB - The neocentric activity of a constriction located on the long arm of rye 5R chromosome (5RL) was analysed. It is not observed in normal rye but it is unusually stretched in bivalents involving 5RL telosomes in wheat-ditelosomic 5RL addition lines. In 20% of metaphase I cells, the 5RL bivalent presents the centromeres oriented to one pole and the constrictions oriented towards the opposite pole with a strong tension. In 5% of the cells, the constriction was able to orient the bivalent to the poles without tension in the centromeres. Sister chromatid cohesion, which is one of the distinct features of centromeric function, is persistent at the constriction in delayed 5RL chromosomes at anaphase I. Neither the elongation of the constriction nor the neocentric activity was observed at second meiotic division or mitosis. FISH studies showed that the 5RL constriction lacked detectable quantities of two repetitive DNA sequences, CCS1 and the 180-bp knob repeat, present at cereal centromeres and neocentromeres, respectively. We propose that, under special conditions, such as the wheat background, the normally non-centromeric DNA present at this region of 5RL acquires a specific chromatin structure, differentiated as an elongated constriction, which is able to function as a centromere. PMID- 11032324 TI - Spatial separation of parental genomes during mouse interspecific (Mus musculus x M. spretus) spermiogenesis. PMID- 11032325 TI - Human COP9 subunit 8 homolog gene SGN8 Map position 2q37. PMID- 11032326 TI - Human NRG3 gene Map position 10q22-q23. PMID- 11032327 TI - Human SS13 gene Map position 17q25.3. PMID- 11032328 TI - Human ALY/BEF gene Map position 17q25.3. PMID- 11032329 TI - Isolated and contiguous glycerol kinase gene disorders: a review. AB - Glycerol kinase deficiency (GKD) is an X-linked recessive disorder. There are two types. an isolated form and a complex form. We review the clinical, biochemical and molecular genetic features of GKD. The clinical and biochemical phenotype of isolated GKD may vary from a life-threatening childhood metabolic crisis to asymptomatic adult 'pseudohypertriglyceridaemia', resulting from hyperglycerolaemia. To date 38 patients from 24 families with isolated GKD have been reported. At least 7 of these patients had a metabolic crisis during a catabolic condition. The complex GKD is an Xp21 contiguous gene syndrome involving the glycerol kinase locus together with the adrenal hypoplasia congenita (AHC) or Duchenne muscular dystrophy (DMD) loci or both. Clinical features of a patient with complex GKD depend on the loci that are involved. Approximately 100 patients from 78 families with a complex GKD have been reported. Seventeen patients with complex GKD (AHC-GKD-DMD or AHC-GKD) died in the neonatal period or early childhood because of unrecognized or inappropriate management of adrenal dysfunction. Since the outcome of the crisis in GKD is highly dependent on the physicians' knowledge of the disease, we devised an algorithmic approach to the diagnosis. From molecular genetic investigations of isolated GKD, 7 missense mutations, 2 splice site mutations, I nonsense mutation, 1 Alu Sx insertion and 2 small deletions were reported for isolated GKD in 13 unrelated families. In 4 families consisting of more than one patient with the same biochemical and genetic defect, the phenotypic variability of the isolated GKD was remarkable. The clinical variability in isolated GKD cannot be explained by biochemical or by molecular heterogeneity. Isolated GKD patients showed a tendency towards hypoglycaemia with hyperketonaemia; whether the clinical symptoms of GKD are caused by dysfunction of gluconeogenesis and/or ketolysis needs to be investigated further. PMID- 11032330 TI - Behaviour and school achievement in patients with early and continuously treated phenylketonuria. AB - Thirty patients with early and continuously treated phenylketonuria (PKU) between 8 and 20 years of age were compared with 30 controls, matched individually for age, sex, and educational level of both parents, on behaviour rating scales for parents and teachers as well as a school achievement scale. PKU patients, as a group, demonstrated more problems in task-oriented behaviour and average academic performance than did matched controls. Interestingly, whereas male PKU patients were rated significantly lower on introversion by their teachers, female patients were rated significantly higher on introversion and lower on extraversion than matched controls. This sex difference was also reflected in the relationship between measures of dietary control and the behaviour clusters, suggesting that male and female patients respond differently to elevated Phe levels or the stress associated with PKU. The teacher rating on average academic performance of the PKU patients was associated with recent level of dietary control, which suggests that it might be improved by more strict adherence to the diet. In addition, academic performance correlated negatively with the behaviour cluster negative task orientation. Further studies are recommended to obtain a more complete evaluation of this relationship and to replicate the current findings on larger samples. Over the years a number of studies have examined behaviour and school achievement in patients with early treated phenylketonuria (PKU; McKusick 261600). In general, these studies have found that despite early treatment with a phenylalanine (Phe)-restricted diet, PKU patients demonstrate more behavioural and school problems than do healthy controls. The behaviour problems include both internalizing symptoms (e.g. solitary, unresponsive, anxious, depressed mood: Pietz et al 1997; Smith et al 1988; Weglage et al 1992) and externalizing symptoms (e.g. hyperactive, talkative, impulsive, restless: Hendrikx et al 1994; Kalverboer et al 1994; Realmuto et al 1986; Smith et al 1988), but not antisocial or socially negative symptoms (e.g. lying, teasing, disobedience: Kalverboer et al 1994; Pietz et al 1997; Smith et al 1988). With respect to school achievement, studies have shown that patients with early treated PKU more often repeat classes or need special tutoring (Berry et al 1979; Brunner et al 1983; Koch et al 1987; Rey et al 1996; Verkerk 1995), have to work harder than healthy controls to achieve the same results (Weglage et al 1993), or have specific deficits in arithmetic achievement scores (Azen et al 1991; Berry et al 1979; Fishler et al 1987; Koch et al 1987; Weglage et al 1993). Nevertheless, many questions regarding the behavioural and school problems of patients with early treated PKU remain unanswered. For instance, the relationship between behavioural and school problems on the one hand and levels of dietary control on the other is still relatively unclear. The few studies that examined this relationship, have focused primarily on children in primary school (Azen et al 1991; Koch et al 1987; Smith et al 1988). Furthermore, although several psychological studies have shown that the pattern of behavioural problems varies by sex (see Prior et al 1999a for a discussion), so far very few studies have examined this issue in PKU patients and results are contradictory (Kalverboer et al 1994; Pietz et al 1997; Smith et al 1988; Weglage et al 1992). In addition, so far no study has actually examined whether there is a relationship between the behavioural problems and school difficulties of PKU patients, even though this relationship has been well documented in the psychological literature (Prior et al 1999b; Richards et al 1995). The aim of the present study is therefore to examine these issues in patients with early and continuously treated PKU over a wide age range and in relation to dietary control. More specifically, school achievement as well as social and task-oriented behaviour (at home PMID- 11032331 TI - Clinical significance of brain phenylalanine concentration assessed by in vivo proton magnetic resonance spectroscopy in phenylketonuria. AB - Recent studies using in vivo proton magnetic resonance spectroscopy (1H MRS) have suggested that plasma phenylalanine (Phe) may not be a reliable indicator of brain Phe level in subjects with phenylketonuria (PKU). Interindividual variation in cerebral Phe can contribute to the phenotypic variability of the disease. We report the results of the direct assessment of brain Phe by 1H MRS in 10 off-diet PKU patients (aged 15.5-30.5 years), 4 detected and treated early, 6 late. In a single patient, brain Phe was evaluated before and 15 days after diet discontinuation. FLAIR MRI and 1H MRS were performed in the same setting by a 1.5 T clinical MR scanner. MR images were scored according to the extent of the lobar white-matter hyperintensity. Brain 1H MRS Phe signal (resonating at 7.36 ppm) was evaluated as a ratio to the creatine+phosphocreatine signal. Brain Phe was correlated with clinical, biochemical and MRI findings. Results were as follows. (1) An abnormal concentration of brain Phe was detected in all 10 PKU subjects (ranging from 0.030 to 0.074), associated with a wide interindividual variability of concurrent plasma Phe (ranging from 724 to 2800 micromol/L). (2) In late detected subjects, brain Phe concentration correlated with clinical phenotype better than did plasma Phe. The discrepancy between brain and plasma Phe was relevant from a clinical point of view in two cases: in one, a late-detected patient with normal mental development, a high level of plasma Phe was associated with a relatively low concentration of brain Phe; in the other, a late-detected subject with severe neurological impairment, a very high level of brain Phe was associated with plasma Phe compatible with the diagnosis of mild PKU. (3) White matter alterations were detected in all patients. FLAIR MRI sequences disclosed an involvement of optic chiasma and tracts in 7 subjects. No correlation was found between white-matter alterations and concurrent brain Phe concentrations. (4) In the only case assessed under different intake of Phe, the relevant increase of brain Phe paralleled the concurrent increase of plasma Phe, showing that 1H MRS can be a useful tool in evaluating the individual vulnerability of PKU patients to different values of plasma Phe. PMID- 11032332 TI - Acylcarnitines in plasma and blood spots of patients with long-chain 3 hydroxyacyl-coenzyme A dehydrogenase defiency. AB - The acylcarnitines in plasma and blood spots of 23 patients with proven deficiency of long-chain 3-hydroxyacylcoenzyme A dehydrogenase were reviewed. Long-chain 3-hydroxyacylcarnitines of C14:1, C14, C16 and C18:1 chain length, and long-chain acylcarnitines of C12, C14:1, C14, C16, C18:2 and C18:1 chain length were elevated. Acetylcarnitine was decreased. In plasma, elevation of hydroxy C18:1 acylcarnitine over the 95th centile of controls, in combination with an elevation of two of the three acylcarnitines C14, C14:1 and hydroxy-C16, identified over 85% of patients with high specificity (less than 0.1% false positive rate). High endogenous levels of long-chain acylcarnitines in normal erythrocytes reduced the diagnostic specificity in blood spots compared with plasma samples. The results were also diagnostic in asymptomatic patients, and were not influenced by genotype. Treatment with diet low in fat and high in medium-chain triglyceride decreased all disease-specific acylcarnitines, often to normal, suggesting that this assay is useful in treatment monitoring. PMID- 11032333 TI - Quantitative analysis of glucose-6-phosphate translocase gene expression in various human tissues and haematopoietic progenitor cells. AB - We investigated the quantitative expression of the human glucose-6-phosphate translocase gene (G6PT1) and its splicing variants in human tissues. The G6PT1 gene was strongly expressed in liver, kidney and haematopoietic progenitor cells, which might explain major clinical symptoms such as hepatomegaly, nephromegaly and neutropenia in glycogen storage diseases type Ib. Reverse transcriptase mediated PCR amplification of G6PT1 cDNA revealed several splicing variants in tissue-specific manners. The brain-specific isoform, which has an additional 22 amino acids between exons 6 and 8, was also identified in heart and skeletal muscle. A new splicing variant, although less prominent in quantity and lacking polypeptide loops corresponding to exons 2 and 3, may have a distinct substrate affinity or specificity in leukocytes and haematopoietic progenitors. In conclusion, the G6PT1 gene was expressed in various tissues, and alternative splicing variants exist in tissue-specific manners. PMID- 11032334 TI - Isolation and characterization of the normal canine beta-galactosidase gene and its mutation in a dog model of GM1-gangliosidosis. AB - The acid beta-galactosidase cDNA of Portuguese Water dogs was isolated and sequenced. The entire coding region of the gene consists of 2004 nucleotides encoding a protein of 668 amino acids. Its encoding sequence indicates approximately 86.5% identity at the nucleotide level and about 81% identity at the amino acid level with the encoding region of the human acid beta galactosidase gene. The deduced amino acid sequence contains a 24-amino-acid putative signal sequence, six possible glycosylation sites, and seven cysteine residues. A homozygous recessive mutation, causing canine GM1-gangliosidosis, was identified at nucleotide G200-->A in exon 2 resulting in an Arg60-->His (mutation R60H) amino acid substitution. The mutation creates a new restriction enzyme site for Pml1. Genotyping 115 dog samples for this acid beta-galactosidase gene alteration readily distinguished affected homozygous recessives (n=5), heterozygous carriers (n=50) and normal homozygotes (n=60). DNA mutation analysis provided a method more specific than enzyme assay of beta-galactosidase for determination of carriers. PMID- 11032335 TI - Lovastatin does not correct the accumulation of very long-chain fatty acids in tissues of adrenoleukodystrophy protein-deficient mice. AB - Lovastatin, an inhibitor of 3-hydroxy-3-methylglutarylcoenzyme A reductase, normalizes the very long-chain fatty acids (VLCFA) concentrations in fibroblasts and plasma from patients with X-linked adrenoleukodystrophy (X-ALD). The effects of lovastatin on the accumulation of VLCFA in tissues of adrenoleukodystrophy protein (ALDP)-deficient mice were assessed. ALDP-deficient mice were fed chow with 0.01-0.1% lovastatin for 4-8 weeks. The VLCFA concentrations in the plasma, brain, spinal cord, liver and kidneys were measured. Treatment with 0.1% lovastatin significantly reduced body weight and total cholesterol in the plasma of ALDP-deficient mice. Treatment with lovastatin, however, did not correct the accumulation of VLCFA in the plasma or tissues, including the brain and spinal cord. Lovastatin does not affect the accumulation of VLCFA in ALDP-deficient tissues in mice. PMID- 11032336 TI - Rolipram does not normalize very long-chain fatty acid levels in adrenoleukodystrophy protein-deficient fibroblasts and mice. AB - In its severe form, X-linked adrenoleukodystrophy (X-ALD) is a lethal neurodegenerative disorder with inflammatory demyelination, in which defective peroxisomal beta-oxidation causes accumulation of very long-chain fatty acids (VLCFA) in tissues and plasma, in particular in the nervous system and adrenal glands. Recently, several drugs have been reported to reduce VLCFA in cultured human fibroblasts of X-ALD patients, and therefore to be potential candidates for novel therapeutic treatments in X-ALD. Among the most promising of these substances is the antidepressant rolipram, because of favourable adverse event profile in clinical studies and its additionally reported anti-inflammatory action. To further elucidate the effects of rolipram on peroxisomal beta oxidation and VLCFA accumulation, we administered rolipram orally in the diet to ALD protein-deficient mice and ALD protein-deficient cultured human and mouse fibroblasts and assayed the accumulation of VLCFA. In contrast to the previously reported reduction of VLCFA, our data did not demonstrate a decrease in VLCFA content either in vivo or in vitro. NMR spectroscopic analysis verified the structural integrity and purity of the rolipram used here, thus excluding inauthenticity as a reason for the discrepancy. We therefore suggest that rolipram should be excluded from the current list of potential therapeutic agents for X-ALD. PMID- 11032337 TI - Biochemical and molecular basis for mitochondrial cardiomyopathy in neonates and children. AB - Defects in myocardial bioenergetics have been reported in patients with cardiomyopathy but their molecular basis and role in pathophysiology remain unclear. We sought to establish a molecular basis for cardiac mitochondrial respiratory enzyme abnormalities frequently present (75%) in a group of 16 children (including 2 neonates) with end-stage cardiomyopathy. Decreased specific activity levels were found in complexes I, III, IV and V but not in II, the only complex that is entirely nuclear encoded. Sequence analysis of cardiac mtDNA revealed 4 patients harbouring heteroplasmic mtDNA mutations in cytb, tRNAArg, and ND5 at highly conserved positions. These mutations were present neither in controls nor in patients without enzymatic defect. In addition, 4 patients exhibited marked reduction in cardiac mtDNA levels. The basis for respiratory enzyme abnormalities can be explained in a subset of our patients as a result of either pathogenic mtDNA mutation or depletion. Patients harbouring both DNA and enzymatic defects fulfil rigorous criteria defining mitochondrial cardiomyopathy. PMID- 11032338 TI - False positive fructose loading: a pitfall in the diagnosis of fructose-1,6 bisphosphatase deficiency. PMID- 11032339 TI - Cerebral infarction and pancreatitis: possible complications of patients with 3 hydroxy-3-methylglutaryl-CoA lyase deficiency. PMID- 11032340 TI - Smith-Lemli-Opitz syndrome with extremely low plasma cholesterol. PMID- 11032341 TI - Myosin heavy chain composition of the human lateral pterygoid and digastric muscles in young adults and elderly. AB - The myosin heavy chain (MyHC) content in different parts of, two jaw opening muscle, the human lateral pterygoid and the digastric muscles of five young adult and five elderly subjects (mean age 22 and 73 years, respectively) was determined, using gel electrophoresis and immunohistochemical methods. The lateral pterygoid of both young and elderly contained predominantly slow MyHC, and fast A MyHC was the major fast isoform. In contrast, the digastric was composed of slow, fast A and fast X MyHCs in about equal proportions in both age groups. About half of the lateral pterygoid fibres contained mixtures of slow and fast MyHCs, often together with alpha-cardiac MyHC. In the digastric, co existence of slow and fast MyHCs was rare, and alpha-cardiac MyHC was lacking. On the other hand, co-expression of fast A and fast X MyHCs was found more often in the digastric than in the lateral pterygoid. In both age groups about half of the digastric IIB fibres contained solely fast X MyHC. In the lateral pterygoid, type IIB fibres with pure fast X MyHC was found in only one subject. The lateral pterygoid in elderly showed a significant amount of fibres with solely fast A MyHC, which were occasionally found in young adults. In the digastric, no significant differences were found between young and elderly, although the muscles of elderly contained lower mean value of slow MyHC, as compared to that of young muscles. It is concluded that the lateral pterygoid and the digastric muscles differ not only in the MyHC composition but also in modifications of the MyHC phenotypes during aging, suggesting that they have separate roles in jaw opening function. PMID- 11032343 TI - Binding of filamin isoforms to myofibrils. AB - Two filamin isoforms were purified from bovine tissues and characterized. Muscle filamin and nonmuscle filamin had different SDS gel mobilities, proteolytic digestion patterns, myofibrillar binding distributions and myofibril binding affinities. The muscle specific filamin had an apparent molecular weight of 265 kDa and bound primarily to the Z-lines of myofibrils but also to the I-bands near the Z-lines. The nonmuscle specific filamin had an apparent molecular weight of 275 kDa and bound exclusively to the Z-lines of myofibrils. The filamin myofibril binding was studied quantitatively. Plotting bound fraction (mg filamin/mg myofibril) vs. equilibrium concentration of free filamin yielded a biphasic binding curve. The first hyperbolic binding phase described the binding of filamin to myofibrils but the second phase appeared to be nonspecific due to filamin aggregation. The muscle filamin had a significantly lower (P < 0.05) apparent binding affinity to myofibrils than nonmuscle filamin. However, the muscle filamin showed a significantly higher (P < 0.05) saturation value for myofibrils than nonmuscle filamin. The binding of phosphorylated filamin to myofibrils was significantly lower (P < 0.05) than the corresponding native proteins for both filamin isoforms. PMID- 11032342 TI - Length-dependent effects of osmotic compression on skinned rabbit psoas muscle fibers. AB - The goal of this study was to characterize the interrelationship between sarcomere length and interfilament spacing in the control of Ca2+ sensitivity in skinned rabbit psoas muscle fibers. Measurements were made at sarcomere lengths 2.0, 2.7 and 3.4 microm. At 2.7 microm the fiber width was reduced by 17% relative to that at 2.0 microm and the pCa50 for force development was increased by approximately 0.3 pCa units. In the presence of 5% Dextran T-500 the fiber width at sarcomere length 2.0 microm was also decreased by 17% and the Ca2+ sensitivity was increased to the same value as at 2.7 microm. In contrast, at sarcomere length 2.7 microm the addition of as much as 10% Dextran T-500 had no effect on Ca2+ sensitivity. At sarcomere length 3.4 microm there was an additional 7% compression and the Ca2+ sensitivity was increased slightly (approximately 0.1 pCa units) relative to that at 2.7 microm. However at 3.4 microm the addition of 5% Dextran T-500 caused the Ca2+ sensitivity to decrease to the level seen at 2.0 microm. Given that the skinning process causes a swelling of the filament lattice it is evident that the relationship between sarcomere length and Ca2+ sensitivity observed in skinned fibers may not always be applicable to intact fibers. These data are consistent with measurements of Ca2+ in intact fibers which indicate that there might be a decline in Ca2+ sensitivity at long sarcomere lengths. PMID- 11032344 TI - A thermodynamic muscle model and a chemical basis for A.V. Hill's muscle equation. AB - Direct measurements of a relationship between force and actin-myosin biochemistry in muscle suggest that molecular forces in active muscle rapidly equilibrate among. not within, individual myosin crossbridges [Baker et al. (1999) Biophys J 77: 2657 2664]. This observation suggests a thermodynamic model of muscle contraction in which muscle, not an individual myosin crossbridge, is treated as a near-equilibrium system. The general approach can be applied to any ensemble of molecular motors that undergo a physicochemical step against a constant external potential. In this paper we apply the model to a simple two-state crossbridge scheme like that proposed by A.F. Huxley (1957) [Prog Biophys 7: 255 317], and we immediately obtain A.V. Hill's muscle equation. We show that this equation accurately describes steady-state muscle mechanics, biochemistry and energetics. This thermodynamic model provides a novel description of force-dependent actin myosin kinetics in muscle and provides precise chemical expressions for myosin cooperativity, myosinduty ratios, the number of working strokes per ATP hydrolyzed, muscle efficiency. and energy transfer. PMID- 11032346 TI - Actomyosin interactions in a novel single muscle fiber in vitro motility assay. AB - A novel in vitro motility assay has been developed to study the actomyosin interaction, in which the molecular motor protein myosin has been extracted and immobilized directly from 2-4 mm single rat skeletal muscle fiber segments. This method study was carried out to investigate: (1) the amount of myofibrillar proteins extracted from the fiber segment; (2) the effects of temperature on the speed at which actin is propelled by fast and slow myosin; and (3) the effects of myosin isoform expression on motility speed. Approximately 80% of the myosin and myosin-associated proteins were extracted from the fiber segments. while no significant extraction was shown of the thin filament proteins. Fluorescently labeled actin filaments moved with constant speed in a bi-directional motion over the high-density myosin region in the experimental chamber, and motility speed was highly dependent on the myosin heavy chain (MyHC) isoform extracted. At 25 degrees C, significant (P < 0.001) differences in motility speed were obtained between type I (1.31 +/- 0.23 microm/s, n = 11) and IIxb (5.81 +/- 0.35 microm/s, n = 6), or llb (6.07 +/- 0.33 microm/s, n = 8) MyHC isoforms. The motility speed and maximum velocity of unloaded shortening (V0) in single fibers were well correlated, indicating that filament speed is a good molecular analogue to contractile speed at the fiber level. The effects of temperature on filament motility speed were analyzed from 10 to 35 degrees C. The Q10 values, calculated in the 10-25 degrees C temperature range, differed between slow (4.20) and fast (2.38) myosin. In conclusion, this in vitro motility assay offers a unique possibility to compare the regulatory and modulatory influence of myosin isoforms and thin filament proteins on shortening velocity, at the cellular and molecular level in the same muscle fiber. PMID- 11032345 TI - Human skeletal myosin heavy chain genes are tightly linked in the order embryonic IIa-IId/x-ILb-perinatal-extraocular. AB - Myosin heavy chain (MyHC) is the major contractile protein of muscle. We report the first complete cosmid cloning and definitive physical map of the tandemly linked human skeletal MyHC genes at 17p13.1. The map provides new information on the order, size, and relative spacing of the genes. and it resolves uncertainties about the two fastest twitch isoforms. The physical order of the genes is demonstrated to contrast with the temporal order of their developmental expression. Furthermore, nucleotide sequence comparisons allow an approximation of the relative timing of five ancestral duplications that created distinct genes for the six isoforms. A firm foundation is provided for molecular analysis in patients with suspected primary skeletal myosinopathies and for detailed modelling of the hypervariable surface loops which dictate myosin's kinetic properties. PMID- 11032347 TI - The maximal velocity of vascular smooth muscle shortening is independent of the expression of calponin. AB - In smooth muscle, the phosphorylation/dephosphorylation of the 20-kDa regulatory light chain of myosin (MLC20) is known to regulate actomyosin interaction and force. However, a thin filament based regulatory system for actomyosin interaction has been suggested to exist in parallel to MLC20 phosphorylation. Calponin is a thin filament associated protein that in vitro inhibits actomyosin interaction, and has been suggested to reduce maximal shortening velocity (vmax). Using antibodies to h1- and h2-calponin, we demonstrated that calponin was present in smooth muscle from Sprague Dawley (SD) rats, while calponin was not detectable in the smooth muscle from Wistar Kyoto (WKY) rats. vmax determined from the force vs. velocity relationship at maximal Ca2+ activation was not different for either the aorta or the portal vein of SD vs. WKY rats. These results suggest that physiological levels of calponin do not contribute to a thin filament-based secondary regulation to inhibit smooth muscle contraction. PMID- 11032348 TI - Functional diversity between orthologous myosins with minimal sequence diversity. AB - To define the structural differences that are responsible for the functional diversity between orthologous sarcomeric myosins, we compared the rat and human beta/slow myosins. Functional comparison showed that rat beta/slow myosin has higher ATPase activity and moves actin filaments at higher speed in in vitro motility assay than human beta/slow myosin. Sequence analysis shows that the loop regions at the junctions of the 25 and 50 kDa domains (loop 1) and the 50 and 20 kDa domains (loop 2), which have been implicated in determining functional diversity of myosin heavy chains, are essentially identical in the two orthologs. There are only 14 non-conservative substitutions in the two myosin heavy chains, three of which are located in the secondary actin-binding loop and flanking regions and others correspond to residues so far not assigned a functional role, including two residues in the proximal S2 domain. Interestingly, in some of these positions the rat beta/slow myosin heavy chain has the same residues found in human cardiac alpha myosin, a fast-type myosin, and fast skeletal myosins. These observations indicate that functional and structural analysis of myosin orthologs with limited sequence diversity can provide useful clues to identify amino acid residues involved in modulating myosin function. PMID- 11032350 TI - Higher target haemoglobin level and early anaemia treatment: different or complementary concepts? AB - There is little doubt that epoetin is a highly effective treatment for renal anaemia. However, it has been used primarily to treat dialysis patients, in whom there is good evidence that it induces significant improvements in cardiac function, exercise capacity, and quality of life. Unfortunately, none of these three major parameters is completely normalized. There are three possible reasons for this: (i) the anaemia is not fully corrected, (ii) too much damage has already occurred by the time the patient starts dialysis, and (iii) other contributory factors may be playing a part. Although the effects of completely correcting renal anaemia have been examined in various studies, the results have not been as positive as expected. It therefore seems appropriate to consider a new strategy in which epoetin therapy is initiated at an earlier stage in the course of the disease, e.g. at a haemoglobin concentration of 11 g/dl or less. It is possible, for example, that earlier treatment of anaemia could prevent many cardiac problems and other morbidities in renal patients. In addition, if epoetin therapy is started in patients who have not been exposed to long-term chronic anaemia, fewer complications may be encountered when reversing the anaemia. Higher target haemoglobin concentrations may also be appropriate in these patients. It would certainly be inappropriate, however, to extrapolate the data on normalization of haemoglobin in dialysis patients to the pre-dialysis population. It is therefore necessary to re-examine the issue of optimal target haemoglobin concentration in pre-dialysis patients. One of the challenges in the new millennium must be to better understand the consequences of initiating treatment of anaemia earlier in the course of chronic renal failure. PMID- 11032349 TI - Cross-bridge movement and stiffness during the rise of tension in skeletal muscle -a theoretical analysis. AB - Predictions for the time courses of cross-bridge attachment. N(t), stiffness, S(t), and force, T(t), during the tetanus rise were analysed for a special class of cross-bridge models where cross-bridges initially attach in a non stereospecific weak-binding state, AW. This state is in rapid equilibrium (equilibrium constant K) with detached states and the force generating transition (rate constant F+) is delayed. One model (model IA) which assumed step-function rise of activation at onset of tetanus, gave a poor fit to the experimental data (judged by root mean square error, RMSe approximately 0.038) but the experimentally observed lead of N(t) over T(t) was reproduced qualitatively. An activation mechanism where K increased towards its maximum value according to an exponential function (Model IB) improved the fit considerably (RMSe approximately 0.013). However, the activation time constant (r = 30 ms) derived in the fit was too high to reflect Ca2+ binding to troponin. In a further developed model (model II) both Ca2+ -binding to troponin and cross-bridge attachment were assumed to be required for full activation. This more complex model gave a good fit to the experimental data (RMSe approximately 0.013) with a realistic time constant for Ca2+ binding to troponin (9 ms). In both model IB and model II the best fit was obtained with F+ approximately 40 s(-1). An extended version of model IB, with distributed cross-bridge attachment and a series elastic element, gave a fit of similar quality (RMSe approximately 0.009) as obtained with model IB and model II and with a similar value of F+. The results support the view that weakly bound cross-bridges (state AW) may account for the lead of cross-bridge movement over force during tension rise. It is also shown that, if the stiffness of the myofilaments is non-linear (stiffness increasing with tension) the experimentally observed lead of S(t) over T(t) may, to a significant degree, be attributed to cross-bridges in the state AW. PMID- 11032351 TI - Consequences of late referral on patient outcomes. AB - There is growing awareness of a need not only to identify patients with chronic renal failure (CRF) at an earlier stage in the disease process, but also to initiate treatment strategies earlier, in order to delay both progression of CRF and co-morbid diseases and to define the optimal time required to prepare CRF patients for renal replacement therapy (RRT). These three strategies are linked, and rely on appropriate identification of patients at risk of renal disease. The challenge currently facing nephrologists is both how to minimize the consequences of late referral and how to improve the timeliness of referral. Published studies support the notion that outcomes are poor in patients who access specialized nephrology care late in the course of their renal disease (just prior to the need for dialysis). A National Institute of Health consensus publication recommends early referral to a multidisciplinary renal care team, and the recent Canadian Society of Nephrology guidelines recommend that at least 12 months are needed prior to initiation of dialysis for adequate medical and psychological preparation for RRT. Despite these recommendations, a substantial proportion (20 50%) of patients starts dialysis without prior exposure to nephrologists. Limited data exist on current referral patterns to nephrologists. Diabetes and/or hypertension cause renal disease in up to 40% of patients requiring dialysis. These patients are presumably being monitored by internists, endocrinologists or cardiologists, and many referrals come from these physicians; other patients may be referred by general practitioners. Data regarding disease status at the time of referral are also limited. Substantial cardiovascular disease and risk factors are evident at the time of referral. Most of the literature describes data for those starting dialysis (i.e. late referral) rather than a broader spectrum of all patients with renal insufficiency referred to nephrologists. Reasons for late referral include insensitivity of current screening tools. Serum creatinine is well known to be an inaccurate marker of renal dysfunction, and too insensitive to identify patients with very early stages of disease, thus contributing to the prevalence of late referrals. Physician and patient attitudes are other barriers to early referral and need to be studied more fully. The consequences of late referrals include increased morbidity, mortality, and resource utilization. There is also an impact on patients' quality of life and missed opportunities for pre emptive transplantation. Late referral also limits therapeutic options, and these limitations have consequences on long-term outcomes once patients are on dialysis. It is clear that late referral of patients with CRF obviates the opportunity for significant delay of disease progression and institution of proactive strategies to reduce the overall burden of illness in the population. There is ample evidence that strategies to delay progression of renal disease are effective, as are strategies to reduce cardiovascular disease. Anaemia and a fall in haemoglobin concentration have been associated with left ventricular hypertrophy and with growth of the left ventricle. A combined approach is necessary for best nephrological clinical practice, with a clear definition of early renal insufficiency; this will involve the development of tools to permit early identification of patients with early renal insufficiency, and the implementation of strategies to optimize treatments aimed at both delaying progression and preparing patients for RRT. PMID- 11032352 TI - Pathophysiology of anaemia: focus on the heart and blood vessels. AB - The amount of oxygen delivered to an organ depends on three factors: blood flow and its distribution; the oxygen-carrying capacity of the blood, i.e. haemoglobin concentration; and oxygen extraction. Non-haemodynamic and haemodynamic mechanisms operate to compensate for anaemia. Non-haemodynamic mechanisms include increased erythropoietin production to stimulate erythropoiesis, and increased oxygen extraction (displacement of the haemoglobin oxygen dissociation curve). This decreased affinity of oxygen for haemoglobin is mediated by increased 2,3 diphosphoglycerate concentrations. Increased cardiac output is the main haemodynamic factor, mediated by lower afterload, increased preload, and positive inotropic and chronotropic effects. Decreased afterload is due to vasodilatation and reduced vascular resistance as a consequence of lower blood viscosity, hypoxia-induced vasodilatation, and enhanced nitric oxide activity. Vasodilatation also involves recruitment of microvessels and, in the case of chronic anaemia, stimulation of angiogenesis. With decreased afterload, the venous return (preload) and left ventricular (LV) filling increase, leading to increased LV end-diastolic volume and maintenance of a high stroke volume and high stroke work. High stroke work is also due to enhanced LV contractility attributed to increased concentrations of catecholamines and non-catecholamine inotropic factors. In addition, heart rate is increased in anaemia, due to hypoxia-stimulated chemoreceptors and increased sympathetic activity. In the long term, these haemodynamic alterations lead to gradual development of cardiac enlargement and LV hypertrophy (LVH). The LVH is eccentric, characterized by increased LV internal dimensions and a normal ratio of wall thickness to cavity diameter, as occurs in other forms of volume overload. When anaemia-related LVH develops in an otherwise 'healthy' humoral environment, the lesions are reversible and the type of LVH is primarily physiological and is not associated with impaired diastolic function. In the absence of underlying cardiovascular disorders, severe anaemia (Haemoglobin concentration < 4-5 g/dl) leads to congestive heart failure. In the presence of heart disease, especially coronary artery disease, anaemia intensifies angina and contributes to a high incidence of cardiovascular complications. In end-stage renal disease (ESRD), LVH is influenced by many other factors, leading to intense interstitial fibrosis, to alterations in diastolic function, and usually to poor reversibility. The chronic increase in cardiac output contributes to arterial remodelling of central elastic arteries such as the aorta and common carotid artery. This remodelling consists principally of arterial enlargement and compensatory arterial intima--media thickening. In ESRD, these geometric changes are accompanied by arterial stiffening. The principal consequences of arterial alterations are increased systolic pressure and high inertia due to higher blood mass in the dilated arterial system. These alterations contribute to the development of LVH and abnormal coronary perfusion. PMID- 11032353 TI - Effects of anaemia on cardiovascular status. AB - Cardiomyopathy is a common, heterogeneous and important cause of cardiac morbidity and mortality in uraemic patients. The risks of ischaemic heart disease, cardiac failure, and death increase progressively from lowest risk in patients with concentric left-ventricular hypertrophy, to medium risk in patients with left-ventricular dilatation but intact systolic function, to highest risk in patients with systolic dysfunction. Anaemia and hypertension are the reversible risk factors most consistently linked with the development of cardiomyopathy in these patients. Longitudinal data show that anaemia predisposes individuals to initial left ventricular dilatation, with compensatory hypertrophy, which may progress to systolic dysfunction. This process typically begins at glomerular filtration rates between 25 and 50 ml/min, and haemoglobin concentrations that are even slightly below normal are associated with progressive cardiac enlargement. Several observational studies have suggested that the correction of anaemia may reduce mortality and hospitalization rates in dialysis patients. The available evidence supports maintaining haemoglobin concentrations to greater than 11 g/dl. Whether a haemoglobin threshold exists above which no further benefit is seen remains controversial, partially because recent randomized controlled trials have intervened relatively late in the anaemia cardiomyopathy cardiac failure death continuum. One large randomized controlled trial showed no benefit from normalizing the haemoglobin concentration in haemodialysis patients with well-established cardiac disease; however, these patients had been exposed to anaemia for long periods of time and were at the extreme end of the cardiorenal disease spectrum. Other researchers have demonstrated a protective effect of normalizing the haemoglobin concentration in patients with asymptomatic, and hence presumably early, cardiomyopathy. The psychological benefits and improvements in exercise tolerance and quality of life resulting from normalization of the haemoglobin concentration are becoming clearer. However, conclusive evidence of the cardiovascular benefits of earlier, more aggressive treatment of renal anaemia as well as of the exact target haemoglobin concentration at which risk begins to develop is still lacking. The results of ongoing trials should help to clarify both of these issues within the next 5 years. PMID- 11032354 TI - Quality of life benefits of early anaemia treatment. AB - In evaluating outcomes in end-stage renal disease (ESRD), quality of life has become as important as morbidity and mortality. Various instruments are available to analyse patients' perceptions of the physical, psychological and social domains of health. Non-specific instruments, such as the Sickness Impact Profile, the Karnofsky Scale, and the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36), have been widely used in evaluating quality of life in various chronic diseases including ESRD. The Kidney Disease Quality of Life (KDQOL) questionnaire and other scales have also have been developed specifically for ESRD patients. Several studies have demonstrated a significant improvement in quality of life after initiation of epoetin treatment in both dialysis patients and those with early renal failure. Quality-of-life scores show a strong positive correlation with haemoglobin concentration. Other factors associated with better quality of life are higher socio-economic level and level of education. However, older age, comorbidity, diabetes, female sex, and unemployment have a negative influence on quality of life. In patients not yet on dialysis, quality of life deteriorates as the glomerular filtration rate decreases. The later the patient is referred to a nephrologist, the worse the quality of life. Recent studies show that quality of life is a prognostic factor for survival. Early and effective treatment of anaemia in ESRD patients is essential in maintaining quality of life both before and after initiation of dialysis. PMID- 11032355 TI - Benefits of anaemia treatment on cognitive function. AB - The role of anaemia in the pathogenesis of abnormal cognitive function in haemodialysis patients is only partly understood. Several contemporary studies have assessed cognitive function in haemodialysis patients using neuropsychological and neurophysiological tests, comparing the results obtained over a wide range of haematocrits before and after treatment with epoetin. Of the neurophysiological tests, which measure electrical impulses from the brain in the presence or absence of specific stimuli, the cognitive event-related potential (ERP) has been particularly useful in evaluating this issue. Changes in amplitude (increase) and latency (decrease) of a specific wave form (P300) seen in the ERP correlate with improved cognitive function in haemodialysis patients following initiation of dialysis therapy. Similar effects are also evident when haematocrit is increased. Marsh et al. (Kidney Int 1991; 39: 155 163) examined the effect of increasing haematocrit with epoetin therapy on the P300 amplitude and latency, and noted that after increasing haematocrit to 23.7 36% over 12 months with epoetin, P300 amplitude increased (P<0.025), and several neuropsychological test scores were significantly improved. Grimm et al. (Kidney Int 1990; 38: 480 486) found a significant decrease in P300 latency and statistically nonsignificant improvements in P300 amplitude when haematocrit was increased to 22.7 30.6%. Improvement in the electroencephalogram (EEG) and P300 latency has also been reported by others, including Sagales et al. (Kidney Int 1993; 44: 1109-1115). The effect of complete reversal of anaemia (haematocrits 31.6 42.9%) on cognitive function was recently revealed by Pickett et al. (Am J Kidney Dis 1999; 33: 1122 1130), who found improvement in several cognitive ERP manoeuvres and a significant decrease in EEG slowing (P< 0.02) at higher haematocrits. Taken together, these various studies suggest that graded increments in haematocrit following epoetin therapy have a progressive effect on improving cognitive function in haemodialysis patients. PMID- 11032356 TI - Management of early renal anaemia: diagnostic work-up, iron therapy, epoetin therapy. AB - Effective management of early anaemia in the course of chronic renal insufficiency requires the following: (i) implementing an efficient diagnostic strategy to exclude common contributing factors; (ii) initiating epoetin therapy for the majority of patients; for and (iii) ensuring adequate iron supply erythropoiesis. Diagnostic inquiry is warranted whenever the haemoglobin concentration is below the normal range adjusted for age and gender. The most efficient diagnostic approach is to assume erythropoietin deficiency, exclude iron deficiency, and pursue further diagnostic tests only when red-cell indices are abnormal or when leukopenia or thrombocytopenia are also present. Macrocytosis should prompt an inquiry into alcoholism, B12 deficiency, or folate deficiency. Microcytosis suggests iron deficiency or thalassaemia. Associated cytopenias raise the possibility of alcohol toxicity, pernicious anaemia, malignancy, or myelodysplastic syndrome. Epoetin therapy is warranted whenever the haemoglobin concentration has fallen below 10.0 g/dl. To initiate therapy prior to dialysis, epoetin should be administered at an average dose of 100 IU/kg/week (80-120 IU/kg/week, 50-150 IU/kg/ week) by subcutaneous injection. Haemoglobin concentration should be monitored every 2 weeks and the epoetin dose adjusted by increments or decrements of 25% to maintain a rate of rise of haemoglobin concentration of 0.2-0.6 g/dl (0.3 0.6 g/dl/week, 0.2-0.5 g/dl/week). When the target range is achieved, the dose of epoetin should be continually adjusted to maintain a stable haemoglobin concentration. Transferrin saturation and ferritin concentration should be monitored monthly, and sufficient iron provided to maintain transferrin saturation above 20%. The lower the haemoglobin concentration, the greater the likelihood that future intravenous iron will be required. Oral iron supplements should be avoided, since they are costly, ineffective, and troublesome to patients. Finally, a blunted therapeutic response to epoetin therapy provides important diagnostic information and gnostic inquiry. PMID- 11032357 TI - Early epoetin treatment in patients with renal insufficiency. AB - Historically, epoetin has been used to treat anaemia in patients already receiving renal replacement therapy. For many years, however, the results of early animal experiments raised considerable concern among nephrologists that disease progression would be accelerated if epoetin therapy were initiated in the predialysis phase of renal failure. In retrospect, it has become clear that the results of these early animal experiments were confounded by a concomitant and uncontrolled rise in blood pressure. In subsequent studies in rat models, antihypertensive treatment effectively prevented the adverse effect on disease progression. In addition, the results of several small observational studies and one large controlled study suggest that the glomerular filtration rate is not adversely affected in pre-dialysis patients treated with epoetin as long as blood pressure is well controlled. There are several observations, though not definitive, which suggest that disease progression may even be slower when anaemia is reversed. The benefits of early anaemia treatment with epoetin include increased exercise capacity and improved quality of life, cognitive function, and sexual function. Anaemia has also been identified as an important aetiological factor in the development of left ventricular hypertrophy. Whether pre-emptive treatment of anaemia is indicated in all pre-dialysis patients, or at least in those who develop progressive left ventricular hypertrophy, is currently under investigation. PMID- 11032358 TI - Potential use of isolated glomeruli and cultured mesangial cells as in vitro models to assess nephrotoxicity. AB - The purpose of this short review is to present the potential of using isolated glomeruli and cultured mesangial cells as two different in vitro models to assess the glomerular effect of molecules with nephrotoxic properties. The advantage of using isolated renal glomeruli is that they conserve the architecture of this anatomical region of the kidney; moreover, they are free of any vascular, nervous or humoral influences derived from other regions of the kidney. Mesangial cells are perivascular pericytes located within the central portion of the glomerular tuft between capillary loops. Mesangial cells have a variety of functions including synthesis and assembly of the mesangial matrix, endocytosis and processing of plasma macromolecules, and control of glomerular hemodynamics, mainly the ultrafiltration coefficient Kf, via mesangial cell contraction or release of vasoactive hormones. Most authors agree that mesangial cells play a major role in glomerular contraction, filtration surface area, and Kf regulation. One of the major effects of toxicants on glomerular structures is contraction. We can assess quantitatively the degree of toxicant-induced mesangial cell contraction or glomerular contraction by measuring the changes in planar cell surface area or apparent glomerular cross-sectional area after exposition to the toxicant. These in vitro models can also reveal glomerular effects of xenobiotics that are difficult or impossible to observe in vivo. In addition, these studies permit a fundamental examination of the mechanism of action of xenobiotics on glomerular cells, including the possibility that at least a part of their effects are mediated by local mediators released by glomerular cells. We review the effects and the mechanisms of action of several toxicants such as gentamicin, cyclosporin, cisplatin, and cadmium on isolated glomeruli or cultured mesangial cells. As such in vitro results confirm in vivo renal hemodynamic changes caused by toxicants, we conclude that these models are fruitful tools for the study of renal toxicity. These in vitro systems might also serve as a predictive tool in the evaluation of drugs inducing changes in glomerular filtration rate and as a way to propose protective agents against these dramatic hemodynamic effects. PMID- 11032359 TI - U373-MG response to interleukin-1beta-induced oxidative stress. AB - Oxidative stress has been involved in various neurological disorders and, in the central nervous system, astrocytes represent the cell type that contributes to neuroprotection via glutathione (GSH) metabolism, GSH-metabolizing enzymes like gamma-glutamyltransferase (GGT), and apoE secretion. In this study, using IL 1beta, a proinflammatory and prooxidant cytokine that is increased in numerous pathological situations, cells of astrocytoma cell line U373-MG were exposed to an oxidative stress, leading to c-Jun and c-Fos activation. IL-1beta decreased both GGT activity and intracellular GSH content and increased apoE secretion, initiating astroglial response to injury. We observed that antioxidants inhibit IL-1beta effects on c-Jun and c-Fos proteins, GGT activity and the GSH pool but not on apoE secretion. Our results allow us to conclude that neurological disorders associated with an IL-1beta-induced oxidative stress could be, at least experimentally, reversible in the presence of one antioxidant, N-acetylcysteine. PMID- 11032360 TI - Dehydroepiandrosterone and melatonin prevent Bacillus anthracis lethal toxin induced TNF production in macrophages. AB - The lethal toxin of Bacillus anthracis, which is composed of two separate proteinaceous exotoxins, namely protective antigen and lethal factor, is central to the pathogenesis of anthrax. Low levels of this toxin are known to induce release of cytokines such as tumor necrosis factor alpha (TNF-alpha). In the present study we investigated the effect of dehydroepiandrosterone (DHEA), melatonin (MLT), or DHEA + MLT on production of lethal toxin-induced TNF-alpha in mouse peritoneal macrophages. We found that treatment with DHEA significantly inhibited the TNF-alpha production caused by anthrax lethal toxin. Exposure of MLT to anthrax lethal toxin-treated macrophages also decreased the release of TNF alpha to the extracellular medium as compared to the control. However, combined use of DHEA and MLT also inhibited TNF-alpha release, but not more than single therapies. These results suggest that DHEA and MLT may have a therapeutic role in reducing the increased cytokine production induced by anthrax lethal toxin. PMID- 11032361 TI - Adaptive response is differently induced depending on the sensitivity to radiation-induced cell death in mouse epidermal cells. AB - We investigated the relationship between induction of radio-adaptive response and cell death in mouse normal and neoplastic epidermal cells. Mouse normal primary keratinocytes (PK), cancer-prone cells [v-rasHa-transfected mouse keratinocytes (ras-PK), and line 308 cells (mouse skin papilloma cells which have activated rasHa gene with A-to-T transversion at codon 61) were primed with a low dose of gamma-rays (0.01 Gy), and were challenged with a high dose (4 Gy) after a 4 or 7 h interval. The induction of cell death in PK was 2-10 times higher and was also more rapid in PK than in ras-PK or 308 cells. Low-dose pretreatment with a 4 h interval decreased cell death, and this adaptive response was prominent in PK, whereas it was less obvious in the cases of ras-PK and 308 cells. The response of each protein kinase C (PKC) isozymes to high-dose radiation, especially PKCalpha, PKCdelta, PKCepsilon, and PKCeta, were different between the normal and ras oncogene-activated neoplastic keratinocytes; translocation of these isozymes to membrane occurred more rapidly in normal than in neoplastic cells. Furthermore, low-dose pretreatment did not induce the translocation of PKCdelta in PK significantly more than in ras-PK and 308. Thus, the difference in the induction of radio-adaptive responses between mouse normal and neoplastic epidermal cells reflects difference in the rapidity of cell death, and responsiveness of PKC may affect this adaptive response. PMID- 11032363 TI - Effects of extracts from sporoderm-broken spores of Ganoderma lucidum on HeLa cells. AB - The effects of extracts from Ganoderma lucidum spores on the growth of human cervix uteri tumor HeLa cells as well as on the cell cycle and intracellular calcium level were investigated. Alcohol extracts were prepared from sporoderm broken and sporoderm-nonbroken spores (termed extract I and extract II) of G. lucidum. Extract I was then subjected to silica gel chromatography to obtain extract III. Cytotoxicity was examined by means of trypan blue exclusion and MTT tests. It was found that extract I and extract III, but not extract II strongly inhibited the growth of HeLa cells, and that extract III was more effective than extract I. Moreover, extract III was shown to be capable of blocking the cell cycle at the transition from G1 to S phase and inducing a marked decrease of intracellular calcium level, determined by flow cytometry and the specific fluorescent calcium probe Fura-2, respectively. These results imply that (1) the breaking of G. lucidum spores improves the release of cytotoxic activity and (2) the effective extract might influence the cell cycle and cellular signal transduction by altering the calcium transport system. PMID- 11032362 TI - Induction of apoptosis in normal cultured rat hepatocytes and in Hep3B, a human hepatoma cell line. AB - The in vitro occurrence of apoptosis in hepatic cells has not been well characterized because it depends on apoptosis inducing-agents and culture conditions. Furthermore, for a given hepatic cell and the same agent, discrepant results have been reported depending on the technique used to evaluate the proportion of apoptotic cells. In this study, we compared the effects of several apoptosis-inducing agents - transforming growth factor beta1 (TGF-beta1), retinoic acid (RA), okadaic acid (OA), and cycloheximide (CY) - on two types of hepatic cells, the human hepatoma cell line Hep3B and normal rat hepatocytes, maintained either plated for 24 to 48 h or in suspension for 20 h. Chromatin condensation and/or nucleus fragmentation were investigated morphologically by DAPI staining. DNA fragmentation was investigated biochemically by agarose gel electrophoresis and poly(ADP-ribose) polymerase (PARP) cleavage was studied by western blot. Apoptotic cells were quantified either by counting cells on UV microscopy after DAPI staining or by flow cytometry. Nuclear changes, the ladder pattern on DNA electrophoresis and PARP cleavage were observed in plated cells, hepatoma cells and normal rat hepatocytes, with all inducers but especially with OA. Semiquantification confirmed that OA was a strong inducer in plated cells under the present conditions, since about 14% and 30% of Hep3B cells (with DAPI staining and flow cytometry, respectively) were apoptotic after 48 h treatment, while, with the other inducers, apoptosis was weaker and discrepancies were also observed between the two counting methods (TGF-beta1; 4% and 12%; RA, 7% and 12%; CY, 4% and 16%, with DAPI staining and flow cytometry, respectively). OA induced a moderate apoptosis in cultured hepatocytes (13% with DAPI staining), while TGF beta1, RA and CY were found to be weakly apoptotic (respectively 4% for the first two and 6% for the last ) after 48 h. In contrast, in suspension cells, apoptosis was observed neither in Hep3B cells nor in normal hepatocytes, whatever the apoptotic inducer and whatever the techniques used to detect apoptosis. In conclusion, our results show that induction of apoptosis in hepatic cells depends not only on the apoptosis-inducing agent but also on the culture conditions. PMID- 11032364 TI - Transformation of the tropane alkaloid-producing medicinal plant Hyoscyamus muticus by particle bombardment. AB - We report an efficient whole plant transformation system for Hyoscyamus muticus, an important medicinal plant of the Solanaceous family. We developed a system using a plasmid carrying the nptII and gusA genes, which was delivered into leaf explants by particle bombardment. Ten percent of bombarded leaf explants formed kanamycin-resistant callus, from which putative transgenic plants were recovered. The nptII gene conferring kanamycin resistance was found to be incorporated into the genome of all transgenic plants screened. Over 50% of the kanamycin resistant plants showed strong expression of the non-selected gusA gene. The majority of transgenic plants reached maturity, could be self pollinated, and produced fertile seed. A simple and efficient whole plant transformation system for this medicinal plant is an important step in furthering our understanding of tropane alkaloid production in plants. PMID- 11032365 TI - Adaptation of Spodoptera exigua (Lepidoptera: Noctuidae) to barley trypsin inhibitor BTI-CMe expressed in transgenic tobacco. AB - Nicotiana tabacum plants were transformed with the cDNA of barley trypsin inhibitor BTI-CMe under the control of the 35S CaMV promoter. Although the transgene was expressed and the protein was active in the homozygous lines selected, growth of Spodoptera exigua (Lepidoptera: Noctuidae) larvae reared on transgenic plants was not affected. The protease activity in larval midgut extracts after 2 days feeding on transformed tobacco leaves from the highest expressing plant showed a reduction of 25% in the trypsin-like activity compared to that from insects fed on non-transformed controls. The susceptibility of digestive serine-proteases to inhibition by BTI-CMe was confirmed by activity staining gels. This decrease was compensated with a significant induction of leucine aminopeptidase-like and carboxipeptidase A-like activities, while chymotrypsin-, elastase-, and carboxipeptidase B-like proteases were not affected. PMID- 11032366 TI - Direct transformation and plant regeneration of the haploid liverwort Marchantia polymorpha L. AB - Thalli of the haploid liverwort Marchantial polymorpha were successfully used for direct particle bombardment with plasmid pMT, which carries a hygromycin phosphotransferase gene (hpt) controlled by the CaMV 35S promoter and the NOS polyadenylation region. Hygromycin-resistant cell masses arose from the thallus surface and developed directly into hygromycin-resistant thalli. Southern blot analyses indicated that these thalli carried at least 1-4 copies of the hpt gene, which were stably transmitted to their asexual thallus progenies via gemma propagation for three generations. This transformation and direct plant regeneration protocol is expected to be a valuable tool for the molecular analysis of this lower land plant. PMID- 11032367 TI - A murine monoclonal antibody produced in transgenic plants with plant-specific glycans is not immunogenic in mice. AB - Previous studies have shown that the production of recombinant antibodies in plants is highly efficient and presents numerous therapeutic applications. It is, however, known that plant glycoproteins display different glycosylation patterns to those exhibited by mammalian glycoproteins. Thus, it is important to know if these plant recombinant antibodies could induce undesirable immune responses in mammals; and to date no report has documented the potential immunogenicity of parenterally administered plant recombinant antibodies in animals. In order to answer this question, mice were immunised subcutaneously with a recombinant mouse monoclonal antibody produced in tobacco plants, together with alum as adjuvant. Two control groups were immunised in the same way with either the original murine monoclonal antibody or horseradish peroxidase (a plant glycoprotein). Analyses by direct immunoassay, competition immunoassay and real-time surface plasmon resonance, showed undetectable levels of antibody directed against both the protein and the glycan part of the plant recombinant antibody. These results have a direct relevance for the application of plant recombinant proteins as therapeutic agents and vaccines in humans. PMID- 11032368 TI - Ribozyme-mediated resistance to rice dwarf virus and the transgene silencing in the progeny of transgenic rice plants. AB - A hammerhead ribozyme (Rz) with long hybridizing arms targeting the mRNA of rice dwarf virus (RDV) segment 5 and a mutated nonfunctional ribozyme (mRz) were constructed. As predicted, Rz transcribed in vitro cleaved the target mRNA of RDV segment 5 into two fragments of 138 and 238 nucleotides in length. The Rz and mRz genes were each placed under the control of the CaMV 35S promoter and used to transform Japonica rice variety 'Tongling No. 1' via Agrobacterium tumefaciens. A total of 32 independent lines containing Rz or mRz was obtained as demonstrated by Southern blot analysis. Challenge inoculation with RDV viruliferous leafhoppers (Nephotettix cincticeps) showed that T1 plants containing the Rz transgene displayed high resistance or delayed and attenuated viral symptoms. In contrast, transgenic lines expressing mRz showed severe symptoms similar to the control plants transformed with the vector alone. These results suggest that Rz confers RDV resistance in transgenic rice. Genomic DNA PCR analysis confirmed that all of the examined T6 progeny plants contained the Rz transgene. However, accumulation of the Rz transcripts was detectable by RT-PCR only in the plants that were resistant to RDV. This suggested that loss of RDV resistance in progeny plants containing the Rz transgene may result from silencing of the Rz transgene. PMID- 11032369 TI - An in vivo model of hyperacute rejection: characterization and evaluation of the effect of transgenic human complement inhibitors. AB - Hyperacute rejection (HAR) occurring after transplantation within phylogenetically distant species is a severe reaction triggered by preexisting xenoreactive antibodies and complement activation, leading to the destruction of the donor organ. Expression of human complement inhibitors in transgenic pig organs prolongs the survival of xenograft in experimental models. Moreover, the extent of protection from hyperacute rejection is dependent on the level and site of expression of the transgenic molecules and, probably, on the combination of different molecules. In this regard a small animal model to test the efficacy of expression vectors and different human molecules could be very advantageous. A murine model developed in our laboratory was characterized by measurement of several parameters characteristic of HAR in the livers of control and transgenic mice expressing transgenic human DAF (CD55) or MCP (CD46) at the end of 2 h of perfusion with human plasma and after I day. The parameters studied were heamatological values of hepatic functions (GOT and GPT), induction of pro inflammatory molecules and histopathological evaluation. Cytokines (IL-1alpha, IL 1beta, IL-6) induction and exposure of P-selectin on the endothelial cell surface, was only observed in control animals after 2 h of perfusion, as an early event. GOT and GPT values increase dramatically after 2 h perfusion and 1 day after the treatment according to the histopathological observation of liver damage. On the contrary, the livers of hDAF or hMCP transgenic mice, under the same treatment were significantly protected although the extent of this protection is dependent on the level of expression of transgenic human molecules. PMID- 11032370 TI - Production of biologically active human granulocyte colony stimulating factor in the milk of transgenic goat. AB - We have developed a transgenic female goat harboring goat beta-casein promoter/human granulocyte colony stimulating factor (G-CSF) fusion gene by microinjection into fertilized one-cell goat zygotes. Human G-CSF was produced at levels of up to 50 microg/ml in transgenic goat milk. Its biological activity was equivalent to recombinant human G-CSF expressed from Chinese hamster ovary (CHO) cell when assayed using in vitro HL-60 cell proliferation. Human G-CSF from transgenic goat milk increased the total number of white blood cells in C57BL/6N mice with leucopenia induced by cyclophosphamide (CPA). The secreted human G-CSF was glycosylated although the degree of O-glycosylation was lower compared to CHO cell-derived human G-CSF. PMID- 11032371 TI - Transformation of rice with the Arabidopsis floral regulator LEAFY causes early heading. AB - Onset of flowering, or heading date, is an important agronomic trait of cereal crops such as rice and early-heading varieties are required for certain regions in which rice is cultivated. Since the floral control gene LEAFY from Arabidopsis can dramatically accelerate flowering in dictoyledonous plants, the usefulness of LEAFY for manipulating heading date in rice has been tested. Constitutive expression of LEAFY from the cauliflower mosaic virus 35S promoter caused early flowering in transgenic rice, with a heading date that was 26-34 days earlier than that of wild-type plants. Early flowering was accompanied by a small yield penalty and some panicle abnormality. These observations suggest that floral regulatory genes from Arabidopsis are useful tools for heading date improvement in cereal crops. PMID- 11032372 TI - A fast and efficient method for transiently transfecting ES cells: application to the development of systems for conditional gene expression. AB - Classically, mouse embryonic stem (ES) cells are transfected by electroporation, a method that requires a large number of cells. Here we describe a protocol using a liposome based transfection agent that is a very simple, rapid and cost effective way of transiently transfecting very low numbers of ES cells. We found this method very useful in screening a large number of ES clones when working with inducible expression systems in which at least two elements are required for regulated expression of the gene of interest. After stable transfection of the first component, clones can be easily and rapidly screened for expression of the gene of interest by transiently transfecting the second component of the system using this protocol. PMID- 11032373 TI - lacZ and ubiquitously expressed genes: should divorce be pronounced? PMID- 11032374 TI - Variegation associated with lacZ in transgenic animals: a warning note. PMID- 11032375 TI - Mission: not impossible? Candidate gene studies in child psychiatric disorders. PMID- 11032376 TI - Association study on the DUSP6 gene, an affective disorder candidate gene on 12q23, performed by using fluorescence resonance energy transfer-based melting curve analysis on the LightCycler. AB - We introduced a new genotyping method, fluorescence resonance energy transfer based melting curve analysis on the LightCycler, for the analysis of the gene, DUSP6 (dual specificity MAP kinase phosphatase 6), in affective disorder patients. The DUSP6 gene is located on chromosome 12q22-23, which overlaps one of the reported bipolar disorder susceptibility loci. Because of its role in intracellular signalling pathways, the gene may be involved in the pathogenesis of affective disorders not only on the basis of its position but also of its function. We performed association analysis using a T>G polymorphism that gives rise to a missense mutation (Leu114Val). No evidence for a significant disease causing effect was found in Japanese unipolars (n = 132) and bipolars (n = 122), when compared with controls (n = 299). More importantly, this study demonstrates that melting curve analysis on the LightCycler is an accurate, rapid and robust method for discriminating genotypes from biallelic markers. This strategy has the potential for use in high throughput scanning for and genotyping of single nucleotide polymorphisms (SNPs). PMID- 11032377 TI - News about disturbances of neuronal migration bring views to bipolar disorder. PMID- 11032378 TI - No genetic linkage or association between a functional promoter polymorphism in the monoamine oxidase-A gene and panic disorder. PMID- 11032379 TI - Is there an evolutionary mismatch between the normal physiology of the human dopaminergic system and current environmental conditions in industrialized countries? AB - A large body of evidence has recently defined a field theory known as 'evolutionary mismatch', which derives its attributes largely from the fact that current environmental conditions are completely different from those in which the human central nervous system evolved. Current views on the evolutionary mismatch theory lack, however, any attempts to define which brain areas or neuronal circuits should be mostly involved in coding such misevolved traits and to what extent our neurobiological knowledge can be applied to the topographical localization of a specific psychopathology. In this respect the mesocorticolimbic dopaminergic circuits have long been misconceptualized as simple reward or reinforcement systems. Instead, they motivate and coordinate the functions of the higher brain areas that mediate planning and foresight and direct finalized movement in both animals and humans. These systems make animals intensely interested in exploring the world around them, but by the same means they also make them susceptible to the environmental stimuli that have been sought and consumed. It is has been speculated that the cortical dopamine targets that developed most recently in phylogeny are of particular functional value, and that the mesocorticolimbic dopaminergic system is involved in more complex integrative functions than previously assumed. In the present paper I will argue that some mental disorders may have their deep roots in the evolutionary mismatch between the normal physiology of the mesocorticolimbic dopaminergic system and the current environmental conditions in affluent societies. PMID- 11032380 TI - Leptin suppresses semi-starvation induced hyperactivity in rats: implications for anorexia nervosa. AB - Semi-starvation induced hyperactivity (SIH) occurs in rodents upon caloric restriction. We hypothesized that SIH is triggered by the decline in leptin secretion associated with food restriction. To test this hypothesis, rats, which had established a stable level of activity, were treated with leptin or vehicle via implanted minipumps concomitantly to initiation of food restriction for 7 days. In a second experiment treatment was initiated after SIH had already set in. In contrast to the vehicle-treated rats, which increased their baseline activity level by 300%, the development of SIH was suppressed by leptin. Furthermore, leptin was able to stop SIH, after it had set in. These results underscore the assumed major role of leptin in the adaptation to semi-starvation. Because SIH has been viewed as a model for anorexia nervosa, we also assessed subjective ratings of motor restlessness in 30 patients with this eating disorder in the emaciated state associated with hypoleptinemia and after increments in leptin secretion brought upon by therapeutically induced weight gain. Hypoleptinemic patients ranked their motor restlessness higher than upon attainment of their maximal leptin level during inpatient treatment. Thus, hypoleptinemia might also contribute to the hyperactivity frequently associated with anorexia nervosa. PMID- 11032381 TI - Anxiety in healthy humans is associated with orbital frontal chemistry. AB - The present study examines relationships between regional brain chemistry (as identified by localized in vivo three-dimensional single-voxel proton magnetic resonance spectroscopy (1H-MRS) and anxiety (as measured by the State-Trait Anxiety Inventory) in 16 healthy subjects. The relative concentrations of N Acetyl aspartate, choline, glutamate, glutamine, gamma-aminobutyric acid, inositol, glucose and lactate were measured relative to creatine within six 8-cm3 brain voxels localized to: thalamus, cingulate, insula, sensorimotor, dorsolateral prefrontal, and orbital frontal cortices (OFC) in the left hemisphere. Analysis of variance, across brain regions, chemicals, and high and low anxiety groups, showed a relationship between anxiety and chemical composition of OFC, with high anxiety subjects demonstrating 32% increase in overall chemical concentrations within OFC, as compared to the lower anxiety group (F= 60.8, P < 10(-7)). Other brain regions, including cingulate, showed no detectable anxiety dependence. The combination of the state and trait anxiety was highly correlated with the concentration of OFC chemicals (r2 = 0.98), and N Acetyl aspartate in OFC was identified as the strongest chemical marker for anxiety (changed by 43.2% between the two anxiety groups, F = 21.5, P = 0.000005). The results provide direct evidence that the OFC chemistry is associated with anxiety in healthy humans. The method can be used as a neuroimaging/behavioral tool for documentation of OFC chemistry changes in relation to anxiety per se and anxiety disorders. The presented relationship between regional brain chemistry and anxiety reflects the functional/behavioral state of the brain, pointing to possible mechanisms of the neurobiology of anxiety. PMID- 11032382 TI - Sequence and genomic organization of the human G-protein Golfalpha gene (GNAL) on chromosome 18p11, a susceptibility region for bipolar disorder and schizophrenia. AB - The sequence and genomic organization of the human Golfalpha (GNAL) gene were determined. The human GNAL gene was found to contain 12 coding exons, and it spans over 80 kb on chromosome 18p11. 5' RACE analysis suggested an additional transcription initiation start site. Sequence analysis of the putative promoter region revealed conserved binding sites for several transcription factors. Sequence analysis of the 3'-untranslated region revealed the presence of two Alu sequences and two polyadenylation signals. 3' RACE analysis confirmed the functionality of the most downstream poly-a signal. The human GNAL was found to be expressed as a single transcript of about 5.9 kb in the brain. One highly informative dinucleotide repeat was found in intron 5. Additionally, a processed pseudogene for asparagine synthetase was found about 6 kb upstream of the GNAL gene. Knowledge of the sequence and structure of the human GNAL gene provides essential information for further analysis of the GNAL locus at chromosome 18p11 which has been linked to bipolar disorder and schizophrenia. PMID- 11032383 TI - Gene identification using exon amplification on human chromosome 18q21: implications for bipolar disorder. AB - We previously reported linkage between bipolar disorder and a region on human chromosome (HC) 18q21. To identify genes in this region, exon trapping was performed on cosmids isolated from an HC18-specific cosmid library (LL18NC02) using 47 sequence tagged site (STS) markers from 18q21 as hybridization probes. A total of 285 unique sequences (exons) were obtained from 850 sequenced clones. Homology searching of the databases using NCBI's BLAST algorithms revealed that 31 exons have identity to known genes and/or ESTs, seven are identical to regions of finished genomic sequences in the 18q21 region, 20 have significant similarity (>30% sequence identity) to genes from human and/or other species, 19 were repetitive sequences, and 208 sequences (72%) are novel. Seventy per cent of the trapped sequences were predicted to be derived from genes using library screening and RT-PCR analyses. This represents an initial stage in characterizing genes in a susceptibility region for further study in bipolar disorder or other diseases that map to this region. PMID- 11032384 TI - Increased cerebrospinal fluid levels of neurotrophin 3 (NT-3) in elderly patients with major depression. AB - Neurotrophin 3 (NT-3) is a member of the neurotrophin gene family which supports the survival of specific neurons. NT-3 was shown to prevent the death of adult central noradrenergic neurons in vivo, a neuronal population which is associated with the pathophysiology of major depression. We quantitated CSF levels of NT-3 in elderly patients with major depression (DE) and compared them to patients with Alzheimer's disease (AD), and mentally healthy control subjects (CTR). CSF levels of NT-3 were markedly and significantly elevated in the DE group, as compared to either the AD or the CTR group (P < 0.01, and P < 0.001, respectively). In terms of diagnostic accuracy, measurement of NT-3 levels in DE resulted in 73.9% sensitivity, and 89.7% specificity. Increased CSF levels of NT-3 may indicate a disturbance of the central noradrenergic system in patients with DE. NT-3 may constitute a biochemical candidate marker for clinical diagnosis and for the evaluation of therapeutic strategies in DE. PMID- 11032385 TI - Elevated hypothalamic/midbrain serotonin (monoamine) transporter availability in depressive drug-naive children and adolescents. AB - Cumulative data suggest depression in adulthood being connected to reduced availability of brain serotonin while the role of dopamine remains less specific. Prospective studies have shown a continuity of depressive episodes from childhood to adulthood, combined with poor social function and excess mortality. The object of this study was to examine whether alterations in brain serotonin and/or dopamine transporter levels are already present in depressive children and adolescents. We examined 41 drug-naive patients (aged 7-17) by single photon emission tomography (SPET) using iodine-123-labelled 23-carbomethoxy 3P3(iodophenyl) tropane [123I]beta-CIT as a tracer for monoamine transporters. In addition to the ordinary clinical examination, the patients were given a structured interview and information was gathered from teachers and parents with questionnaires. The diagnoses were established by consensus evaluation between three child psychiatrists. To test the serotonin hypothesis and the dopamine hypothesis regarding depression in children and adolescents, the series was divided into groups with depression present (31) and no depression present (10). In this study, the depressive child and adolescent patients had significantly higher serotonin transporter availability (P < 0.02) in the hypothalamic/midbrain area. Age did not correlate to the hypothalamic/midbrain serotonin transporter binding ratio. No significant difference in dopamine transporter availability in striatum was found between the depressive and the nondepressive children and adolescents. PMID- 11032386 TI - A family-based and case-control association study of the dopamine D4 receptor gene and dopamine transporter gene in attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is a highly heritable psychiatric condition of early childhood onset characterised by marked inattention, hyperactivity and impulsiveness. Molecular genetic investigations of ADHD have found positive associations with the 480-bp allele of a VNTR situated in the 3' untranslated region of DAT1 and allele 7 of a VNTR in exon 3 of DRD4. A number of independent studies have attempted to replicate these findings but the results have been inconsistent. We used both family-based and case control approaches to examine these polymorphisms in a sample of 137 children diagnosed with ICD-10, DSM-IV or DSM-III-R ADHD. We found no evidence of association with the DAT1 polymorphism, despite a sample size that has up to 80% power to detect a previously reported effect size. We observed a significant increase in the DRD4 7 repeat allele amongst ADHD probands (21.7%) and their parents (18.9% in mothers, 22.3% in fathers), compared to ethnically matched controls (12.8%). However TDT analysis showed no preferential transmission of allele 7 to ADHD probands. PMID- 11032387 TI - Evidence for linkage of a tandem duplication polymorphism upstream of the dopamine D4 receptor gene (DRD4) with attention deficit hyperactivity disorder (ADHD). AB - Attention deficit hyperactivity disorder (ADHD) is a common childhood-onset neurodevelopmental disorder. Evidence from twin, adoption, and family studies provide support for a genetic contribution to the etiology of ADHD. Several candidate gene studies have identified an association between a 7-repeat variant in exon 3 of the dopamine 4 receptor gene (DRD4) and ADHD. However, in spite of the positive reports finding association of the exon 3 VNTR with ADHD, several other polymorphisms within DRD4 have been identified that conceivably could contribute to risk for ADHD. Recently, another common polymorphism of the DRD4 gene has been described involving a 120-bp repeat element upstream of the 5' transcription initiation site. In this report, we describe results of analysis of the DRD4 120-bp repeat promoter polymorphism in a sample of 371 children with ADHD and their parents, using the transmission disequilibrium test (TDT). Results showed a significant preferential transmission of the 240-bp (long) allele with ADHD. Exploratory analyses of the Inattentive phenotypic subtype of ADHD strengthened the evidence for linkage. These data add further support for the role of DRD4 variants conferring increased risk for ADHD, and imply that additional studies of DRD4 and other related genes are needed. PMID- 11032388 TI - Evidence for the serotonin HTR2A receptor gene as a susceptibility factor in attention deficit hyperactivity disorder (ADHD). AB - A recent study demonstrated that treatment of hyperactive mice with psychostimulants and serotonergic agents produced a calming effect that was dependent on serotonergic neurotransmission and was not associated with any changes in extracellular dopamine levels. The complex interaction between the serotonergic and dopaminergic neurotransmitter systems suggests that a balance between the two systems may be necessary for mediating hyperactive behaviour. Defects in serotonin system genes, therefore, may disrupt normal brain serotonin function causing an imbalance between these neurotransmitter systems leading to the development of attention deficit hyperactivity disorder (ADHD). Using the transmission disequilibrium test (TDT), the current study assesses for linkage disequilibrium between polymorphisms in the serotonin HTR2A receptor gene and ADHD. One hundred and fifteen families with a total of 143 children diagnosed with ADHD (DSM-IV) were genotyped for the His452 Tyr and the T102C polymorphisms in the serotonin HTR2A receptor gene. TDT analysis revealed a preferential transmission of the 452Tyr allele to the affected offspring (P = 0.03), suggesting linkage disequilibrium of this polymorphism with ADHD. This may open a new door in ADHD molecular genetics research, expanding the existing view of a catecholaminergic hypothesis to include a serotonergic hypothesis and should help elucidate the complex interplay among the neurotransmitter systems in the etiology of ADHD. PMID- 11032389 TI - Association of a functional polymorphism of the serotonin transporter gene with anxiety-related temperament and behavior problems in children: a longitudinal study from infancy to the mid-teens. AB - A polymorphism in the promoter region of the serotonin transporter gene (5 HTTLPR) has been associated with anxiety-related personality traits in adults. Initial studies showed that the short allele was associated with higher neuroticism, anxiety and harm avoidance. However, most attempts to replicate these findings have been negative. Because the association of candidate polymorphisms with behavioral traits may vary with stage of development, we investigated the association using participants in a longitudinal study of childhood temperament. DNA was available for 660 children who had been assessed for temperament from 4-8 months to 15-16 years, and for behaviour problems from 3 4 years to 15-16 years. No significant associations were found at most ages. However, at ages 13-14 years and 15-16 years, the long/long genotype was associated with higher anxiety. These findings do not support an association of the short allele with anxiety-related traits in early life. PMID- 11032390 TI - Attention-deficit hyperactivity disorder and the gene for the dopamine D5 receptor. AB - A recent study has suggested a possible association of a polymorphism near the dopamine D5 receptor gene (DRD5) and attention-deficit hyperactivity disorder. The polymorphism studied was a (CA)n repeat located in the cosmid containing the D5 receptor gene2 and the allele that was reported to be associated with attention-deficit hyperactivity disorder (ADHD) was the 148-bp allele. In this study we sought to replicate this finding by testing for biased transmission of the alleles at this same polymorphism in a sample of 92 families with an ADHD proband. We did not observe significant evidence for biased transmission of the 148-bp allele, however we did observe biased transmission of two other alleles, the 136-bp allele and the 146-bp allele. For these two alleles the bias was for these two alleles not to be transmitted to the ADHD children. The number of informative transmissions for these two alleles was small, therefore it would be premature to make any conclusions from our study concerning the role of DRD5 in ADHD. PMID- 11032391 TI - The high affinity neurotensin receptor gene (NTSR1): comparative sequencing and association studies in schizophrenia. AB - Neurotensin and its high affinity receptor (NTSR1) localise within dopaminergic neurones in the mesocortical, mesolimbic and nigrostriatal systems and it is now clear that neurotensin can selectively modulate dopaminergic neurotransmission. This has led to the hypothesis that altered neurotensin function contributes to the pathogenesis of schizophrenia and other psychoses. This hypothesis has been supported circumstantially by a number of lines of evidence. (1) Central administration of neurotensin produces effects similar to those produced by the peripheral administration of atypical antipsychotics. (2) Observations of low levels of neurotensin in the CSF of schizophrenics. (3) Reduced numbers of neurotensin receptors in the brains of schizophrenics. Given the above link between neurotensin and dopamine, and the evidence implicating altered neurotensin function in psychosis, we have postulated that DNA sequence variation in neurotensin or its receptors might be associated with schizophrenia. In keeping with this hypothesis, an association has recently been reported between schizophrenia and the gene encoding the neurotensin high affinity receptor (NTSR1). However, caution is required because the associated marker, a tetranucleotide repeat, is located 3 kb away from the 3' end of the gene and there is no evidence that it is functional. Therefore, as a follow-up to our earlier work on neurotensin, we have now sought to test the hypothesis that DNA sequence variants that alter the structure or expression of the NTSR1 gene (VAPSEs) are associated with schizophrenia. However, while we found 14 novel sequence variants in 28 probands with psychosis, none resulted in an amino acid change, and neither direct nor indirect association studies suggested these are involved in susceptibility to schizophrenia. PMID- 11032392 TI - Brain derived neurotrophic factor (BDNF) gene variants association with age at onset and therapeutic response in schizophrenia. AB - Schizophrenia is a heterogeneous disease involving genetic and environmental factors. The frequency of structural brain abnormalities or physical anomalies supports a neurodevelopmental etiology, especially in early onset schizophrenia. Brain-Derived-Neurotrophic-Factor (BDNF) is involved in the neurodevelopment of dopaminergic (DA)-related systems and interacts with the meso-limbic DA systems, involved in the therapeutic response to antipsychotic drugs and substance abuse. In addition, BDNF promotes and maintains dopamine D3 receptor (DRD3) expression. In a French Caucasian population, we found no statistical difference in allele or genotype distribution of the BDNF gene dinucleotide repeat polymorphism (166-174 bp) between the whole group of schizophrenic patients and controls. By contrast, an excess of the 172-176 bp alleles was found in patients with late onset, in neuroleptic-responding patients and in non-substance-abusing patients. BDNF gene variants thus appear to be associated with developmental features of schizophrenia. In addition, this association with good treatment responding was independent from the association found with the DRD3 Ball gene polymorphism in the same population. These results suggest an independent contribution of each gene to a treatment-sensitive form of schizophrenia. PMID- 11032393 TI - Analysis of the disease risk locus DXS1047 polymorphism in Brazilian Alzheimer patients. AB - Alzheimer's disease (AD) is a disorder characterized by a progressive deterioration in memory and other cognitive functions. Four genes associated with early onset AD have been identified but familial AD is rare. The majority of late onset AD (LOAD) is caused by a complex inheritance with several genes interacting with environmental factors. The epsilon4 allele of the apolipoprotein E (APOE) gene has been reported worldwide as a risk factor associated with LOAD. The short variant of a polymorphism in the transcriptional region of the serotonin transporter gene (5-HTTLPR) was analyzed in several psychiatric conditions and found to be more frequently associated with European and Brazilian LOAD patients. Recently, allelic associations with LOAD were reported for five other loci, the most significant for one X-linked 202-bp allele, at the DXS1047 locus. We have analyzed this locus in Brazilian LOAD patients and observed that the 202-bp allele was not significantly more frequent among patients. In contrast, two other alleles (200 bp and 208 bp) were less frequent among AD male patients than in controls, confirming the importance of replicating association studies in different populations. PMID- 11032394 TI - Macrophage migration inhibitory factor (MIF): its essential role in the immune system and cell growth. AB - Macrophage migration inhibitory factor (MIF) functions as a pleiotropic protein, participating in inflammatory and immune responses. MIF was originally discovered as a lymphokine involved in delayed hypersensitivity and various macrophage functions, including phagocytosis, spreading, and tumoricidal activity. Recently, MIF was reevaluated as a proinflammatory cytokine and pituitary-derived hormone potentiating endotoxemia. This protein is ubiquitously expressed in various organs, such as the brain and kidney. Among cytokines, MIF is unique in terms of its abundant expression and storage within the cytoplasm and, further, for its counteraction against glucocorticoids. MIF has unexpectedly been found to convert D-dopachrome, an enantiomer of naturally occurring L-dopachrome, to 5,6 dihydroxyindole. However, its physiologic significance remains to be elucidated. It was demonstrated that anti-MIF antibodies effectively suppress tumor growth and tumor-associated angiogenesis, suggesting that MIF is involved not only in inflammatory and immune responses but also in tumor cell growth. At present, MIF cannot be clearly categorized as either a cytokine, hormone, or enzyme. This review presents the latest findings on the role of MIF in the immune system and in cell growth, with regard to tumorigenesis and wound repair, and discusses its potential functions in various pathophysiologic states. PMID- 11032395 TI - IFN-alpha1a gene is the major variant in the North American population. AB - Thirteen interferon (IFN)-alpha functional genes have been reported. A number of these genes have allelic members (variants). In the case of IFN-alpha1, two variants, IFN-alpha1a and IFN-alpha1b, are known. The variants differ from each other by one base change in the coding region, leading to a single change in amino acid sequence and the presence of a restriction site. We have developed oligonucleotide primers for amplification of IFN-alpha1 gene(s) using polymerase chain reaction (PCR). Genomic DNA, obtained from over 23,000 normal healthy individuals and from four human cell lines, were used as templates in PCR to amplify the IFN-alpha1 gene sequences. The resulting PCR products were analyzed by restriction endonuclease digestion and DNA sequencing to identify the presence of variant sequences. The results show that IFN-alpha1a is predominant in the genomic DNA of the population examined. Among the cell lines studied, IFN-alpha1a is the only variant found in U-937 and Namalwa cells, whereas KG-1 cells have only IFN-alpha1b, and EB-3 cells have both IFN-alpha1a and IFN-alpha1b in the genome. PMID- 11032396 TI - Increased stability of macrophage migration inhibitory factor (MIF) in DU-145 prostate cancer cells. AB - Macrophage migration inhibitory factor (MIF) has been localized to the glandular epithelium of the prostate and stimulates the in vitro growth of prostate epithelial cells. [35S]Methionine labeling of MIF protein was used to determine if prostate cells synthesize and secrete this cytokine. The results demonstrated that the DU-145 prostate cancer cells secrete about twice the amount of a more stable protein compared with normal prostate epithelial cells. To investigate if differences in MIF mRNA levels account for the differences in MIF protein secreted by these cells, mRNA stability was analyzed by [3H]uridine incorporation. Following a 12-h pulse, DU-145 cells were found to contain four times the amount of [3H]uridine-labeled MIF mRNA, and this message exhibited a longer half-life than the message found in normal cells (33 h and 19 h, respectively). Nuclear run-on experiments confirmed that the MIF gene is transcribed at a greater rate (1.8-fold) in the DU-145 prostate cancer cells. This study documents, for the first time, that human prostate epithelial cells synthesize and secrete this cytokine. These results indicate that the increased levels of MIF found in prostate cancer cells is likely due to the increased protein and mRNA stability as exhibited by DU-145 cells. PMID- 11032397 TI - Canine interleukin-13: molecular cloning of full-length cDNA and expression of biologically active recombinant protein. AB - Interleukin-13 (IL-13) regulates immune responses mediated by type 2 T helper lymphocytes (Th2) in the human and mouse. To study the function of this cytokine in the dog, we have isolated a cDNA that encodes the full-length canine IL-13 (CaIL-13) precursor polypeptide of 131 amino acids. CaIL-13 shares significant homology with the IL-13 amino acid sequences of cattle (54.1%), mouse (39.6%), and rat (36.6%) but shares the highest identity with human IL-13 (HuIL-13) (61.8%). The predicted CaIL-13 mature polypeptide of 111 residues was expressed in bacteria, and recombinant CaIL-13 (rCaIL-13) was isolated from inclusion bodies and refolded. rCaIL-13 stimulated the proliferation of TF-1 cells, which are derived from human erythroleukemia cells and respond to IL-13 as well as to a number of other human and murine cytokines. CaIL-13 mRNA was readily detectable by reverse transcriptase-polymerase chain reaction (RT-PCR) in cells from lymph nodes and peripheral blood. The gene sequence and biologically active recombinant protein for CaIL-13 will be useful reagents to determine the role of IL-13 in the regulation of canine immune responses. PMID- 11032398 TI - Interaction of retinoic acid and interferon in renal cancer cell lines. AB - Retinoic acid (RA) can potentiate the antitumor effect of interferons (IFN) in a variety of tumor types, including renal cell carcinoma (RCC). The mechanisms by which RA and IFN increase the antitumor effects in RCC are unknown. We used growth assays and mobility shift assays to examine the effects of combining 13 cis-retinoic acid (CRA) and IFN-alpha (plus IFN-gamma) on proliferation and on the expression of the IFN-specific transcription factor IFN-stimulated gene factor 3 (ISGF3) in RCC cell lines. Combining CRA and IFN-alpha resulted in a significant increase in growth inhibition in four cell lines compared with IFN alpha or CRA alone. Binding of nuclear extracts from RCC cells to an IFN stimulated response element (ISRE) oligonucleotide probe following incubation with IFN-alpha was not increased by CRA but was significantly increased by pretreatment by IFN-gamma in a time-dependent fashion. Proliferation assays showed that sequential addition of IFN-gamma and IFN-alpha significantly increased growth inhibition. IFN-alpha but not IFN-gamma or CRA increased the cellular levels Stat2 and p48 but not Statl. IFN-gamma pretreatment enhanced the upregulation of p48 levels by IFN-alpha. Combining RA and IFN results in additive growth inhibition on RCC cell lines. This increase in growth inhibition is not mediated by increased ISGF3 expression. PMID- 11032399 TI - Cytokine message and protein expression during lung granuloma formation and resolution induced by the mycobacterial cord factor trehalose-6,6'-dimycolate. AB - Trehalose-6,6'-dimycolate (TDM), or cord factor, is a mycobacterial cell wall component that induces granuloma formation and proinflammatory cytokine production in vivo and in vitro. The purpose of this work was to better understand the mechanisms by which TDM promotes lung granuloma formation. This was accomplished by characterizing cytokine mRNA expression during TDM-induced alveolitis culminating in cohesive granuloma development. A single intravenous injection of TDM given to C57BL/6 mice produced lung granulomas that peaked in number 5 days after challenge and were nearly resolved by 14 days. mRNA in whole lung preparations was quantitated by bioluminescent RT-PCR. Tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and IL-6 were significantly elevated during granuloma development and decreased during granuloma resolution. There were no detectable changes in mRNA for interferon-y (IFN-y), IL-2, IL-4, IL 5, IL-10, and IL-12(p40). The level of TNF-alpha protein extracted from lung minces highly correlated with morphologic indices of granulomatous inflammation, indicating that it may be an important modulator of the inflammatory intensity induced by TDM. TDM may interact specifically with macrophages in vivo, as evidenced by induction of TNF-alpha, IL-1beta, and IL-6, but not IFN-gamma, protein in bone marrow-derived macrophages from C57BL/6 mice. TDM may therefore play an important role early in macrophage activation during the host granulomatous response to mycobacteria. PMID- 11032400 TI - Interferon coordinately inhibits the disruption of PML-positive ND10 and immediate-early gene expression by herpes simplex virus. AB - Interferons (IFNs) are important components of the innate immune response, limiting herpes simplex virus (HSV) infection. In recombinant HSV-infected cells, IFN inhibited expression of beta-galactosidase from the immediate-early gene, ICP4, promoter. The extent of inhibition was dependent on IFN dose, IFN type, cell type, and multiplicity of infection (moi). IFN inhibited gene transcription, leading to a complete block in ICP4 promoter-driven gene expression in 90% of cells. The same IFN treatments resulted in an increase in the size and number of nuclear domain 10 (ND10) structures that stained positive by immunofluorescence for the promyelocytic leukemia (PML) protein. In cultures infected at low moi with a recombinant HSV producing ICP4 as a fusion protein with green fluorescence protein, the appearance of green fluorescence in the nucleus coincided with loss of PML-positive ND10 in the same nucleus, even in the rare ICP4-expressing IFN treated cells. IFN-dependent inhibition was nearly complete when the immediate early promoter was in the viral genome but was minimal when the promoter was stably integrated into the cellular genome. These data reveal that IFN can completely block viral gene expression in infected cells and that enhancement of the ND10 structure, which is the site of initiation of HSV replication, correlates with the block in viral gene expression. PMID- 11032401 TI - A classification for the interferon-tau. AB - An attempt has been made to provide a rational organization for the many interferon-tau (IFN-tau) sequences entered in GenBank based on phylogenetic analysis and common amino acid substitutions, which might form the basis for a universal nomenclature scheme. Over the 13 years since these genes were first discovered, large numbers of cDNA and gene sequences have been reported, and there is reason to suspect that representatives of all the major ovine and bovine forms have now been described. The data are consistent with the presence of many genes and also allelic variants in sheep and cattle analogous to what has been observed for the IFN-alpha in the human. Future variants should be easily accommodated into the scheme outlined here. A flexible system of nomenclature, based on that used for HuIFN, is needed to provide a common base for comparison between research done in different laboratories and to assign relative biologic potencies to these molecules. PMID- 11032402 TI - IFN-gamma facilitates release of class II-loaded intracellular pools in trophoblast cells: a novel property independent of protein synthesis. AB - Interferon-gamma (IFN-gamma) is an abortion-inducing factor, yet its effects in such a reaction are subject to various levels of regulation. The trophoblast cell line TROPHO-1 can be induced by IFN-gamma to express mRNA and surface class II major histocompatibility complex (MHC) proteins after 8 and 48 h of stimulation, respectively. Untreated cells, however, show an intracellular accumulation of class II antigens earlier (6 h), indicating the existence of MHC pools in the cystosol independent of any induction. On addition of IFN-y, immunofluorescence, subcellular fractionation, and ELISA experiments showed that class II antigen activity detected in the endosomal compartments of the cells could be measured in the culture supernatants. These soluble class II proteins, when isolated and purified using magnetic bead isolation techniques and tested in SDS-PAGE gel and Western blot experiments, had a molecular weight of 70 kDa. Administration of these molecules to pregnant mice as culture supernatants increased the abortion rate and decreased maternal hematocrit levels, effects that could be immunoabsorbed by anti-I-A(d) monoclonal antibodies (mAb). These results indicate that although surface class II molecules are not expressed on trophoblast cells, they accumulate in endosomal compartments and can be released from the cells on addition of IFN-gamma. This new IFN-gamma property, to mobilize intracellular pools of class II MHC antigens in trophoblast cells independent of de novo protein synthesis and induce their release to the extracellular matrix, is a mechanism that appears to be involved in the fetal rejection process, facilitating priming of the maternal organism against the fetal allograft. PMID- 11032403 TI - Early clearance of circulating hepatitis C virus enhanced by induction therapy with twice-a-day intravenous injection of IFN-beta. AB - To improve the long-term efficacy of interferon (IFN) for treatment of chronic hepatitis C virus (HCV) infection, we proposed induction therapy with twice-a-day IFN-beta injection. This study was intended to clarify the antiviral mechanism. Thirty patients were randomly assigned to two groups: group A (twice-a-day therapy) received 3 MU IFN-beta intravenously (i.v.) twice a day for 2 weeks; group B (once-a-day therapy) received 6 MU of IFN-beta daily. HCV RNA, IFN-beta, alanine aminotransferase (ALT), 2'5'-oligoadenylate synthetase (2'5'-AS) activity, and beta2-microglobulin in serum were compared between the two groups during the first 2 weeks of IFN therapy. The clearance rate of serum HCV RNA in group A (86.7%) was significantly higher than that in group B (13.3%) at day 3 (p = 0.0006). No accumulation of IFN-beta was shown in serum throughout the therapy. The ratio (day 3/day 1) of 2'5'-AS activity was significantly higher in group A. Multivariate analysis indicated twice-a-day IFN-beta injection therapy led to significantly early clearance of circulating HCV. Twice-a-day IFN-beta injection therapy could induce biologically enhanced antiviral activities and be an efficient induction therapy for eradication of HCV. PMID- 11032404 TI - Molecular cloning, expression, and characterization of amorpha-4,11-diene synthase, a key enzyme of artemisinin biosynthesis in Artemisia annua L. AB - In plants, sesquiterpenes of different structural types are biosynthesized from the isoprenoid intermediate farnesyl diphosphate. The initial reaction of the biosynthesis is catalyzed by sesquiterpene cyclases (synthases). In Artemisia annua L. (annual wormwood), a number of such sesquiterpene cyclases are active. We have isolated a cDNA clone encoding one of these, amorpha-4,11-diene synthase, a putative key enzyme of artemisinin biosynthesis. This clone contains a 1641-bp open reading frame coding for 546 amino acids (63.9 kDa), a 12-bp 5'-untranslated end, and a 427-bp 3'-untranslated sequence. The deduced amino acid sequence is 32 to 51% identical with the sequence of other known sesquiterpene cyclases from angiosperms. When expressed in Escherichia coli, the recombinant enzyme catalyzed the formation of both olefinic (97.5%) and oxygenated (2.5%) sesquiterpenes from farnesyl diphosphate. GC-MS analysis identified the olefins as (E)-beta-farnesene (0.8%), amorpha-4,11diene (91.2%), amorpha-4,7(11)-diene (3.7%), gamma-humulene (1.0%), beta-sesquiphellandrene (0.5%), and an unknown olefin (0.2%) and the oxygenated sesquiterpenes as amorpha-4-en-11-ol (0.2%) (tentatively), amorpha-4 en-7-ol (2.1%), and alpha-bisabolol (0.3%) (tentatively). Using geranyl diphosphate as substrate, amorpha-4,11-diene synthase did not produce any monoterpenes. The recombinant enzyme has a broad pH optimum between 7.5 and 9.0 and the Km values for farnesyl diphosphate, Mg2+, and Mn2+ are 0.9, 70, and 13 microM, respectively, at pH 7.5. A putative reaction mechanism for amorpha-4,11 diene synthase is suggested. PMID- 11032405 TI - Structure of the amino terminus of a gap junction protein. AB - Charged amino acid residues in the amino terminus of gap junction forming proteins (connexins) form part, if not all, of the transjunctional voltage sensor of gap junction channels and play a fundamental role in ion permeation. Results from studies of the voltage dependence of N-terminal mutants predict that residues 1-10 of Group I connexins lie within the channel pore and that the N terminus forms the channel vestibule by the creation of a turn initiated by the conserved G12 residue. Here we report that intercellular channels containing mutations of G12 in Cx32 to residues that are likely to interfere with flexibility of this locus (G12S, G12Y, and G12V) do not express junctional currents, whereas a connexin containing a proline residue at G12 (Cx32G12P), which is expected to maintain a structure similar to that of the G12 locus, forms nearly wild-type channels. We have solved the structure of an N-terminal peptide of Cx26 (MDWGTLQSILGGVNK) using 1H 2D NMR. The peptide contains two structured domains connected by a flexible hinge (domain-hinge-domain motif) that would allow the placement of the amino terminus within the channel pore. Residues 1-10 adopt a helical conformation and line the channel entrance while residues 12-15 form an open turn. Overall, there is good agreement between the structural and dynamic features of the N-terminal peptide provided by NMR and the functional studies of the voltage dependence of channels formed by wild-type and N-terminal mutations. PMID- 11032406 TI - Characterization of Cyp2d22, a novel cytochrome P450 expressed in mouse mammary cells. AB - Endogenous steroids and numerous environmental agents have potent effects on mammary development and carcinogenesis. Locally produced cytochrome P450 enzymes that modify such molecules are therefore likely to be important regulators of these processes. Here we describe the characterization of a novel mouse gene, termed Cyp2d22, that is highly expressed in the mammary tumor derived cell line RIII/Prl. Cyp2d22 is expressed at intermediate levels in the weakly tumorigenic cell line RIII/MG, whereas expression is low or absent in all normal mouse mammary epithelial cell lines tested and three C3H mammary tumor derived cell lines. Immunoblot analysis of mouse tissues with highly specific antisera indicates that 2D22 protein levels are most abundant in liver, while intermediate levels of expression are seen in adrenal, ovary, and mammary gland. Immunohistochemical staining of liver sections with these antisera demonstrates that 2D22 is most abundant in the first layer or two of parenchymal cells surrounding the central vein, with virtually no expression detected in periportal cells. Interestingly, sequence similarity and functional data suggest that Cyp2d22 may be the mouse ortholog of human CYP2D6. These observations support the hypothesis that 2D22 mediates a distinct, biologically significant activity in relation to other mouse 2D family members. PMID- 11032407 TI - Injurious mechanical compression of bovine articular cartilage induces chondrocyte apoptosis. AB - A bovine cartilage explant system was used to evaluate the effects of injurious compression on chondrocyte apoptosis and matrix biochemical and biomechanical properties within intact cartilage. Disks of newborn bovine articular cartilage were compressed in vitro to various peak stress levels and chondrocyte apoptotic cell death, tissue biomechanical properties, tissue swelling, glycosaminoglycan loss, and nitrite levels were quantified. Chondrocyte apoptosis occurred at peak stresses as low as 4.5 MPa and increased with peak stress in a dose-dependent manner. This increase in apoptosis was maximal by 24 h after the termination of the loading protocol. At high peak stresses (>20 MPa), greater than 50% of cells apoptosed. When measured in uniaxial confined compression, the equilibrium and dynamic stiffness of explants decreased with the severity of injurious load, although this trend was not significant until 24-MPa peak stress. In contrast, the equilibrium and dynamic stiffness measured in radially unconfined compression decreased significantly after injurious stresses of 12 and 7 MPa, respectively. Together, these results suggested that injurious compression caused a degradation of the collagen fibril network in the 7- to 12-MPa range. Consistent with this hypothesis, injurious compression caused a dose-dependent increase in tissue swelling, significant by 13-MPa peak stress. Glycosaminoglycans were also released from the cartilage in a dose-dependent manner, significant by 6- to 13 MPa peak stress. Nitrite levels were significantly increased above controls at 20 MPa peak stress. Together, these data suggest that injurious compression can stimulate cell death as well as a range of biomechanical and biochemical alterations to the matrix and, possibly, chondrocyte nitric oxide expression. Interestingly, chondrocyte programmed cell death appears to take place at stresses lower than those required to stimulate cartilage matrix degradation and biomechanical changes. While chondrocyte apoptosis may therefore be one of the earliest responses to tissue injury, it is currently unclear whether this initial cellular response subsequently drives cartilage matrix degradation and changes in the biomechanical properties of the tissue. PMID- 11032408 TI - Structural and functional characterization of neuwiedase, a nonhemorrhagic fibrin(ogen)olytic metalloprotease from Bothrops neuwiedi snake venom. AB - A fibrino(geno)lytic nonhemorrhagic metalloprotease (neuwiedase) was purified from Bothrops neuwiedi snake venom by a single chromatographic step procedure on a CM-Sepharose column. Neuwiedase represented 4.5% (w/w) of the crude desiccated venom, with an approximate Mr of 20,000 and pI 5.9. As regards the amino acid composition, neuwiedase showed similarities with other metalloproteases, with high proportions of Asx, Glx, Leu, and Ser. Atomic absorption spectroscopy showed that one mole of Zn2+ and one mole of Ca2+ were present per mole of protein. The cDNA encoding neuwiedase was isolated by RT-PCR from venom gland RNA, using oligonucleotides based on the partially determined amino-acid sequences of this metalloprotease. The full sequence contained approximately 594 bp, which codified the 198 amino acid residues with an estimated molecular weight of 22,375. Comparison of the nucleotide and amino acid sequences of neuwiedase with those of other snake venom metalloproteases showed a high level of sequential similarity. Neuwiedase has two highly conserved characteristics sequences H142E143XXH146XXG149XXH152 and C164I165M166. The three-dimensional structure of neuwiedase was modeled based on the crystal structure of Crotalus adamanteus Adamalysin II. This model revealed that the zinc binding site region showed a high structural similarity with other metalloproteases. The proteolyitc specificity, using the Bbeta-chain of oxidized insulin as substrate, was shown to be directed to the Ala14-Leu15 and Tyr16-Leu17 peptide bonds which were preferentially hydrolyzed. Neuwiedase is a Aalpha,Bbeta fibrinogenase. Its activity upon the Aalpha chain of fibrinogen was detected within 15 min of incubation. The optimal temperature and pH for the degradation of both Aalpha and Bbeta chains were 37 degrees C and 7.4-8.0, respectively. This activity was inhibited by EDTA and 1,10-phenantroline. Neuwiedase also showed proteolytic activity upon fibrin and some components of the extracellular matrix. However, it did not show TAME esterase activity and was not able to inhibit platelet aggregation. PMID- 11032409 TI - Isoaspartate formation and neurodegeneration in Alzheimer's disease. AB - We reviewed here that protein isomerization is enhanced in amyloid-beta peptides (Abeta) and paired helical filaments (PHFs) purified from Alzheimer's disease (AD) brains. Biochemical analyses revealed that Abeta purified from senile plaques and vascular amyloid are isomerized at Asp-1 and Asp-7. A specific antibody recognizing isoAsp-23 of Abeta further suggested the isomerization of Abeta at Asp-23 in vascular amyloid as well as in the core of senile plaques. Biochemical analyses of purified PHFs also revealed that heterogeneous molecular weight tau contains L-isoaspartate at Asp-193, Asn-381, and Asp-387, indicating a modification, other than phosphorylation, that differentiates between normal tau and PHF tau. Since protein isomerization as L-isoaspartate causes structural changes and functional inactivation, or enhances the aggregation process, this modification is proposed as one of the progression factors in AD. Protein L isoaspartyl methyltransferase (PIMT) is suggested to play a role in the repair of isomerized proteins containing L-isoaspartate. We show here that PIMT is upregulated in neurodegenerative neurons and colocalizes in neurofibrillary tangles (NFTs) in AD. Taken together with the enhanced protein isomerization in AD brains, it is implicated that the upregulated PIMT may associate with increased protein isomerization in AD. We also reviewed studies on PIMT-deficient mice that confirmed that PIMT plays a physiological role in the repair of isomerized proteins containing L-isoaspartate. The knockout study also suggested that the brain of PIMT-deficient mice manifested neurodegenerative changes concomitant with accumulation of L-isoaspartate. We discuss the pathological implications of protein isomerization in the neurodegeneration found in model mice and AD. PMID- 11032410 TI - Modification of cysteine residues by N-ethylmaleimide inhibits annexin II tetramer mediated liposome aggregation. AB - A role of cysteine residues in annexin II tetramer (AIIt)'s function was investigated using the sulfhydryl reagent N-ethylmaleimide (NEM). Incubation of AIIt with NEM resulted in a dose-dependent inhibition of AIIt-mediated liposome aggregation and loss of sulfhydryl groups of AIIt. The concentration effecting 50% inhibition was 0.18 mM. The inhibition was observed in all Ca2+ concentrations tested (1-1000 microM). NEM had no effects on liposome aggregation mediated by other annexins (I, III, and IV), indicating that the inhibitory effect caused by NEM modification is specific to AIIt. The NEM-treated AIIt still can bind to liposomes. However, once AIIt was bound to membrane, the cysteine residues were protected from NEM modification. Our results suggest that cysteine residues are critical for AIIt-mediated liposome aggregation. PMID- 11032411 TI - New evidence for the multiplicity of ubiquinone- and inhibitor-binding sites in the mitochondrial complex I. AB - Determination of the number of ubiquinone- and inhibitor-binding sites in the mitochondrial complex I (NADH:ubiquinone oxidoreductase) is a controversial question with a direct implication for elaborating a suitable model to explain the bioenergetic mechanism of this complicated enzyme. We have used combinations of both selective inhibitors and common ubiquinone-like substrates to demonstrate the multiplicity of the reaction centers in the complex I in contrast with competition studies that have suggested the existence of a unique binding site for ubiquinone. Our results provide new evidence for the existence of at least two freely exchangeable ubiquinone-binding sites with different specificity for substrates, as well as for a different kinetic interaction of inhibitors with the enzyme. PMID- 11032412 TI - Free-radical-induced inactivation of lysozyme and carbonyl residue generation in protein are not necessarily associated. AB - The 2,2'-azobis(2-amidinopropane) (AAPH)-induced inactivation and oxidative modification of lysozyme, as determined by the loss of tryptophan-associated fluorescence (TAF) and the increase in dinitrophenylhydrazine-reactive carbonyl groups (CO), were studied in the absence and in the presence of antioxidants. AAPH induced a progressive inactivation of the enzyme and a parallel decrease of its TAF. Both changes were closely correlated (R2 = 0.97); however, the inactivation was only partially associated with an increase in CO. The latter reached maximal values at times half those needed to attain maximal losses in both lysozyme activity and TAF. A stoichiometric comparison reveals that whereas over 74% of the enzyme molecules had lost their activity, only 5% exhibited an increment in CO. CO formation was affected differentially by boldine and trolox. Both antioxidants fully protected against the early inactivation and loss of TAF; however, the increase in CO was completely unaffected by trolox. Exposure of lysozyme to Fe3+/ascorbate induced no loss of activity or TAF, but it led to an accumulation of CO similar to that induced by AAPH. Results indicate that CO formation and lysozyme inactivation are two mechanistically dissociable events and that changes in the former parameter can perfectly occur in the absence of changes in the latter. PMID- 11032413 TI - Effects of trans-resveratrol on copper-dependent hydroxyl-radical formation and DNA damage: evidence for hydroxyl-radical scavenging and a novel, glutathione sparing mechanism of action. AB - Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural product occurring in grapes and various other plants with medicinal properties. The phenolic antioxidant has been identified as a potential cancer chemopreventative agent and its presence in red wine has been suggested to be linked to the low incidence of heart disease in some regions of France. Recently, however, resveratrol was reported to promote DNA fragmentation in the presence of copper ions (K. Fukuhara and N. Miyata, 1998, Bioorg. Med. Chem. Lett. 8, 3187-3192), prompting us to investigate this phenomenon in mechanistic detail. By acting as a reducing agent, resveratrol was found to promote hydroxyl-radical (*OH) formation by DNA-bound Cu(H) ions. However, in the presence of either ascorbic acid or glutathione (i.e., under more physiological conditions), the phenolic lost this property and behaved as an antioxidant. In the ascorbate system, resveratrol had no effect on the rate of *OH formation, but protected DNA from damage by acting as a radical scavenging antioxidant. In contrast, in the glutathione system, resveratrol inhibited *OH formation via a novel mechanism involving the inhibition of glutathione disulfide formation. We have concluded, therefore, that the DNA damaging properties of resveratrol, identified recently by Fukuhara and Miyata, will be of no significance under physiological conditions. To the contrary, we have demonstrated that the phenolic behaves as a powerful antioxidant, both via classical, hydroxyl-radical scavenging and via a novel, glutathione-sparing mechanism. PMID- 11032414 TI - Functional implications of the beta-helical protein fold: differences in chemical and thermal stabilities of Erwinia chrysanthemi EC16 pectate lyases B, C, and E. AB - Colonization of plant tissue by the phytopathogen Erwinia chrysanthemi EC16 is aided by the activities of the pectate lyase isozymes (PLs), which depolymerize the polygalacturonic acid component (PGA) of plant cell walls. The bacterium secretes four pectate lyases (PLa, PLb, PLc, and PLe), two of which, PLc and PLe, have been shown to fold into a similar domain motif, the beta-helix. To understand the rationale behind the evolution and retention of these isoforms, the susceptibilities of pectate lyases B, C, and E to chemical and thermal denaturation and the resulting enzymatic inactivation were examined. With guanidine hydrochloride used as a denaturant, all three pectate lyases denatured with transition midpoint guanidine hydrochloride concentrations (Cm) of 1.3, 1.1, and 1.8 M for PLb, PLc, and PLe, respectively. Lyase activity decreased in direct response to loss of secondary structure in all enzymes. Pectate lyases B and C demonstrated increased enzymatic activity at temperatures above 30 degrees C, with maximal activity observed at 40 degrees C for PLb and 35 degrees C for PLc. Transition midpoints (Tm) as measured by circular dichroism were at 46.9 degrees C for PLb and 44.3 degrees C for PLc, indicating detectable conformational changes accompanying thermal inactivation. Decreased enzymatic activity of PLe was observed at all temperatures above 30 degrees C, and the enzyme was found to possess a Tm at 38.9 degrees C. The data demonstrate structural differences among these enzymes that may be the basis for different enzymatic efficiencies under the potential array of environmental conditions experienced by the bacterium. These differences, in turn, may play a part in the retention of these isozymes as virulence factors, allowing the successful colonization of susceptible plant hosts. PMID- 11032415 TI - Expression of cholesterol 7alpha-hydroxylase restores bile acid synthesis in McArdle RH7777 cells. AB - Bile acid synthesis involves several enzymes and occurs only in liver cells. The first and rate-determining step is catalyzed by cholesterol 7alpha-hydroxylase (cyp7a). McArdle RH7777 hepatoma cells do not synthesize bile acids and do not express the cyp7a gene. A synthetic cyp7a gene was stably expressed in this cell line to determine if restoration of cyp7a activity is sufficient to reconstitute the bile acid synthetic pathway. The transfected cells contained the recombinant cyp7a mRNA and the corresponding protein. Microsomes from recombinant cells converted cholesterol into 7alpha-hydroxycholesterol, indicating that the recombinant enzyme was active. Radiolabeled bile acids, originated from exogenously supplied radiolabeled cholesterol, were detected in the culture medium of recombinant cells. Thus, expression of cyp7a is sufficient in restoring bile acid synthesis in McArdle RH7777 cells. The results also show that the additional complement of enzymatic activities required to convert cholesterol into bile acids has remained active in this cell line. PMID- 11032416 TI - Thermodynamics of p53 binding to hdm2(1-126): effects of phosphorylation and p53 peptide length. AB - Upon exposure to DNA-damaging agents, the p53 tumor suppressor protein is stabilized and activated, leading to cell cycle arrest, DNA repair, or apoptosis. One of the major factors that regulates the level and the transcriptional activity of p53 is the hdm2 oncoprotein. hdm2 binds to the N-terminal transactivation domain of p53 to block the transcriptional activity of p53 directly. hdm2 also functions as the E3 ligase that ubiquitinates p53 for proteasome degradation. Fluorescence anisotropy was employed to measure directly the binding of hdm2(1-126) to a p53 N-terminal peptide labeled with Oregon Green (an analogue of fluorescein). Phosphorylation of Ser15 and Ser2O did not affect the binding of the p53 peptide to hdm2. Thrl8 phosphorylation, on the other hand, reduced the binding by at least 20-fold. This suggests that phosphorylation of Thr18 could be a regulatory mechanism that disrupts the hdm2-p53 complex, thus activating p53 in response to DNA damage. The effect of p53 peptide length on binding to hdm2 was also measured quantitatively. Interestingly, p53(18-26) exhibits 10-fold higher affinity to hdm2 than do longer peptides (20- or 35-mer). This result may reflect a strong entropic barrier to binding for the longer peptides. PMID- 11032417 TI - Differential induction of sesquiterpene metabolism in tobacco cell suspension cultures by methyl jasmonate and fungal elicitor. AB - Jasmonates are well documented for their ability to modulate the expression of plant genes and to influence specific aspects of disease/pest resistance traits. We and others have been studying the synthesis of sesquiterpene phytoalexins in elicitor/pathogen-challenged plants and have sought to determine if methyl jasmonate (MeJA) could substitute for fungal elicitors in the induction of capsidiol accumulation by tobacco cell cultures. The current results demonstrate that MeJA does in fact induce phytoalexin accumulation, but with a much more delayed induction time course than elicitor. While elicitor treatment induced strong but transient changes in key enzymes of sesquiterpene biosynthesis, sesquiterpene cyclase, and aristolochene/deoxy-capsidiol hydroxylase, MeJA did not. Instead, MeJA caused a protracted induction of cyclase activity and only a low level of hydroxylase activity. MeJA induced the expression of at least two sesquiterpene cyclase genes, including one that had not been observed previously in elicitor-induced mRNA populations. Only a small portion of the total sesquiterpene cyclase mRNA induced by MeJA was associated with polysomal RNA, suggesting that the MeJA treatment imposed both transcriptional and posttranscriptional regulation in tobacco cells. These results are not consistent with MeJA playing a role in orchestrating defense responses in elicitor-treated tobacco cells, but do provide evidence that MeJA induces a subset of genes coding for the biosynthesis of sesquiterpene phytoalexins. PMID- 11032418 TI - Stimulated nonspecific transport of phospholipids results in elevated external appearance of phosphatidylserine in ras-transformed fibroblasts. AB - The content of phosphatidylserine (PS) was found to be increased three times in the plasma membrane outer leaflet of ras-transformed fibroblasts compared to their nontransformed counterparts. In an attempt to determine the mechanisms responsible for the enhanced external appearance of PS, we investigated the activities of aminophospholipid translocase and the nonspecific lipid scramblase. Both transport systems could separately or in combination contribute to PS accumulation in the extracellular leaflet. Aminophospholipid transfer was assessed by measuring the rate of NBD-PS internalization, and scramblase activity was estimated from the internalization of NBD-PC. The results showed that the aminophospholipid transport was inhibited and the nonspecific transport was stimulated in ras-transformed cells. To assess which of these two transport systems was related to elevation of PS external appearance, each of them was submitted to reversible alterations and the content of PS was measured simultaneously. Aminophospholipid translocase activity was inhibited by pyridyldithioethylamine treatment and reversed by reduction with dithiothreitol. Scramblase activity was modulated by a calcium repletion-depletion procedure. Calcium depletion was performed by cell incubation with BAPTA-AM and EGTA as Ca2+ intracellular and extracellular chelators. Restoration of the intracellular Ca2+ was achieved by cell incubation with Ca2+ and Ca2+-ionophore A23187. The results showed that the changes in PS outer appearance did not correlate with the uptake of NBD-PS but were closely related to NBD-PC internalization, suggesting that the nonspecific bidirectional lipid transfer was the major transport system translocating PS to the outer leaflet in ras-transformed cells. PMID- 11032420 TI - Novel nonpeptidic inhibitors of peptide deformylase. AB - A novel series of nonpeptidic compounds structurally related to the known anticholesteremic thyropropic acid were found to inhibit Escherichia coli peptide deformylase (PDF), with IC50 values in the low-micromolar range. Kinetic analysis of [4-(4-hydroxyphenoxy)-3,5-diiodophenyl]acetic acid reveals competitive inhibition, with a Ki value of 0.66 +/- 0.007 microM. A structure-activity relationship study demonstrates that the carboxylate is required for activity, while the distal phenolic function can be methylated without significant effect. Either decreasing the number of iodine atoms on the molecule to one or increasing the number of iodine atoms to four results in the loss of an order of magnitude in potency. These compounds are the first nonpeptidic inhibitors disclosed and represent a template from which better inhibitors might be designed. PMID- 11032419 TI - Regulation of DNA binding activity of the ligand-activated aryl hydrocarbon receptor by tyrosine phosphorylation. AB - Aryl hydrocarbon receptor (AhR), a member of the bHLH-PAS family, is a ligand activated transcription factor which plays an important role in normal liver development and in mediating the toxicity of polycyclic and halogenated aromatic hydrocarbon pollutants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin. Phosphorylation is known to regulate the transformation process of unliganded AhR into functionally active AhR/ARNT heterodimer that has high affinity for dioxin responsive elements (DRE) and transactivation activity. Here, we report that DRE binding activity of the AhR is regulated by phosphorylation on the AhR/ARNT complex itself. Studies with specific protein phosphatases indicated that tyrosine phosphorylation is involved in this modulation. In addition, the AhR is phosphorylated at tyrosine residue(s) as determined by anti-phosphotyrosine immunoblot analysis. These results suggest that tyrosine phosphorylation on the AhR is required for its DNA binding activity and may provide mammalian cells with another layer of control mechanism that allows cell type specific and developmental stage specific induction of the AhR target genes. PMID- 11032421 TI - Redox regulation of yeast flavin-containing monooxygenase. AB - The flavin-dependent monooxygenase from yeast (yFMO) oxidizes biological thiols such as cysteine, cysteamine, and glutathione. The enzyme makes a major contribution to the pools of oxidized thiols that, together with reduced glutathione from glutathione reductase, create the optimum cellular redox environment. We show that the activity of yFMO, as a soluble enzyme or in association with the ER membrane of microsomal fractions, is correlated with the redox potential. The enzyme is active under conditions normally found in the cytoplasm, but is inhibited as GSSG accumulates to give a redox potential similar to that found in the lumen of the ER. Site-directed mutations show that Cys 353 and Cys 339 participate in the redox regulation. Cys 353 is the principal residue in the redox-sensitive switch. We hypothesize that it may initiate formation of a mixed disulfide that is partially inhibitory to yFMO. The mixed disulfide may exchange with Cys 339 to form an intramolecular disulfide bond that is fully inhibitory. PMID- 11032422 TI - Regulation of the procine 1,25-dihydroxyvitamin D3-24-hydroxylase (CYP24) by 1,25 dihydroxyvitamin D3 and parathyroid hormone in AOK-B50 cells. AB - The 24-hydroxylase is the enzyme responsible for the first step in the catabolism of 1,25-dihydroxyvitamin D3, the active form of vitamin D. This enzyme was shown to be upregulated by 1,25-dihydroxyvitamin D3 itself and downregulated by parathyroid hormone (PTH). Upregulation of 24-hydroxylase by 1,25 dihydroxyvitamin D3 has been characterized; however, the mechanism by which PTH acts to downregulate 24-hydroxylase expression remains unknown. Here we report the cloning of the porcine 24-hydroxylase, and show that 1,25-dihydroxyvitamin D3 stimulated 24-hydroxylase mRNA and activity are repressed by PTH in AOK-B50 cells, a porcine kidney proximal tubule cell line with stably transfected opossum PTH receptors. Forskolin mimicked the effects of PTH consistent with in vivo data, and suppression by PTH was not due to changes in VDR levels. The first 1400 bp of the 24-hydroxylase promoter were not able to mediate the effects of PTH on a reporter gene. In view of the above findings we concluded that AOK-B50 cells are a suitable model for further studying the mechanism of action of PTH on 24 hydroxylase mRNA. PMID- 11032423 TI - Ethanol enhances the stimulatory effects of lysophosphatidic acid on DNA synthesis but not cell proliferation in human and mouse fibroblasts. AB - Lysophosphatidic acid (LPA), a constituent of serum, is a positive regulator of cell growth, while ethanol (EtOH) has been shown to exert both inhibitory and stimulatory effects on mitogenesis. In this work, we examined possible interactions between the effects of EtOH and LPA on DNA synthesis, cell proliferation, activating phosphorylation of p44/p42 mitogen-activated protein kinases (MAPK), and p70 S6 kinase (p70 S6K) activity. In fibroblasts derived from human or mouse embryo or the skin of healthy human subjects, LPA (1-20 microM) and EtOH (40-80 mM) synergistically stimulated DNA synthesis in a zinc-dependent manner. Nevertheless, EtOH did not modify the stimulatory effect of LPA on the proliferation of human embryonal fibroblasts. In the presence of zinc, EtOH did not affect LPA-induced activating phosphorylation of p42/p44 MAPKs, although an inhibitor of MAPK kinase inhibited the combined effects of LPA and EtOH on DNA synthesis. In contrast, in the presence of zinc, EtOH enhanced the stimulatory effect of LPA on p70 S6K activity. The results indicate that in human fibroblasts, in the presence of zinc, EtOH enhances the stimulatory effects of LPA on DNA synthesis, but not on cell proliferation, by a mechanism probably involving activation of p70 S6K. PMID- 11032424 TI - Subchronic and developmental toxicity studies of n-butyl propionate vapor in rats. AB - Two inhalation studies were conducted to evaluate the possible subchronic and developmental toxic effects of n-butyl propionate. In the subchronic study, Sprague-Dawley rats (15/sex/group) were exposed to 0, 250, 750, or 1500 ppm vapor for 6 h/d, 5 d/wk for 13 wk. Five of the rats per sex per group were held after the final exposure for an 8-wk recovery period. Standard parameters of subchronic toxicity were measured throughout the study, and at the end of exposure and recovery periods, necropsies were performed, organs weighed, and tissues processed for microscopic examination. Exposure did not produce marked treatment related deaths or adversely affect clinical signs, hematology, clinical chemistries, organ weights, or the histology of major visceral organs. The only systemic toxic effects were significant decreases in body weight, body weight gain, and feed consumption that occurred in 1500 ppm group rats. Morphologic changes were limited to the nasal cavity as evidenced by a concentration-related increased incidence and severity of olfactory epithelium degeneration in rats of the 750 and 1500 ppm groups. These degenerative microscopic alterations were primarily confined to the olfactory epithelium within the dorsal portion of the medial meatus, with lesser involvement of the olfactory mucosae overlying the tips of some of the adjacent ethmoturbinates. Both the systemic and nasal cavity effects appeared reversible after exposure ceased. In the developmental toxicity study, pregnant Sprague-Dawley rats (24/group) were exposed to 0, 500, 1000, or 2000 ppm vapor for 6 h/d on gestation d 6-15 and sacrificed on gestation d 20. All treatment-group dams exhibited significant reductions in body weight, body weight gain, and feed consumption. Gestational parameters were equivalent across all groups and there were no treatment-related developmental or teratogenic effects. The no-observed-adverse effects levels (NOAELs) determined for nbutyl propionate were 250 ppm for subchronic toxicity (based on the olfactory epithelium degeneration) and 22000 ppm for developmental toxicity (no developmental effects at top dose tested). Under the conditions of this study, a NOAEL was not determined for maternal toxicity. PMID- 11032425 TI - Two-generation reproduction study by dosing with glutaraldehyde in the drinking water of CD rats. AB - Adult male and female CD rats (F0) were dosed with glutaraldehyde (GA; CAS number 111-30-8) in drinking water at concentrations of 0 (controls), 50, 250, or 1000 ppm for a 10-wk prebreed period and through mating, gestation, and lactation. Resultant F1 offspring, selected to be parents of the next generation, were continued on the same regime from prebreed through lactation. Twenty-eight parental animals per sex per generation for each dose group were evaluated for clinical signs, body weight (absolute and gain), and water and food consumption. The offspring were evaluated for survival and body weight to weaning. Necropsy and light microscopic examination of removed tissues were conducted in all F0 and F1 parents and in 10 offspring/sex/group/generation. Average daily consumptions of GA (as mean +/- SD) for the low, intermediate, and high concentrations were respectively 4.25 +/- 0.87, 17.50 +/- 4.16, and 69.07 +/- 14.58 mg/kg/d for F0 parental males, and 6.68 +/- 0.78, 28.28 +/- 4.09, and 98.37 +/- 11.71 mg/kg/d for F0 parental females. The corresponding values for the F1 parents were 4.53 +/ 1.02, 21.95 +/- 4.88, and 71.08 +/- 16.21 mg/kg/d for males and 6.72 +/- 0.84, 29.57 +/- 5.41, and 99.56 +/- 16.72 mg/kg/d for females. There were no effects on parental fertility and mating performance or on pup viability and litter size in any generation. No apparent treatment-related histopathology was seen in parents or offspring. Parental body weights and body weight gains were significantly reduced at 1000 ppm at a few isolated time periods, particularly during prebreed. Food consumption was significantly reduced at 1000 ppm for F0 and F1 parents during the prebreed and gestation periods, and at 250 ppm for F0 males during prebreed and gestation and F1 females during gestation and lactation. Water consumption by the F0 and F1 parents of the 250 at 1000 ppm groups was reduced throughout the prebreed period. At 1000 ppm, average litter weights were reduced over lactation d 21-28 for the F1 and F2 offspring. The no-observed-effect level (NOEL) for adult toxicity was 50 ppm and for offspring 250 ppm. There were no indications of reproductive toxicity, and the NOEL for this study was therefore > 1000 ppm. PMID- 11032426 TI - Pharmacokinetic disposition and tissue distribution of bisphenol A in rats after intravenous administration. AB - This study examined the dose-linearity pharmacokinetics of bisphenol A, a U.S. Environmental Protection Agency (EPA) classified endocrine disruptor, in rats following iv administration. Upon iv injection of 0.2, 0.5, 1, or 2 mg/kg, serum levels of bisphenol A declined biexponentially, with mean initial distribution and elimination half-life ranges of 4-8.2 min and 38.6-62.2 min, respectively. There were no significant alterations in the systemic clearance rate (mean range 90.1-123.6 ml/min/kg) and the steady-state volume of distribution (mean range 4.6 6.0 L/kg) as a function of the administered dose. In addition, the area under the serum concentration-time curve linearly rose as the dose was increased. In a second study, bisphenol A was given by simultaneous iv bolus injection plus infusion to steady state, and levels were measured in serum and various organs. When expressed in concentration terms (e.g., amount accumulated per gram organ weight), bisphenol A was found predominantly in the lung, followed by kidneys, thyroid, stomach, heart, spleen, testes, liver, and brain. Ratios of the organ to serum bisphenol A concentrations exceeded unity for all the organs examined (ratio range 2.0-5.8) except for brain (ratio 0.75). Given the high systemic clearance and short elimination half-life, bisphenol A is unlikely to accumulate significantly in the rat. PMID- 11032427 TI - Contaminated Department of Energy facilities and ecosystems: weighing the ecological risks. AB - With the ending of the Cold War, the United States and other industrialized nations are faced with the monumental task of cleaning up hazardous waste sites that were created over a 50-year period. While many factors enter into the cleanup decisions of managers of hazardous waste facilities, such as at Department of Energy sites, risk assessment plays a central role in evaluating the hazards to ecosystems and humans. Often risk from hazardous facilities is determined by identifying the risks to people, or immediate ecosystems, without regard to the size or magnitude of the surrounding ecological system or of the receptors within that system. Identifying the types of systems for contaminated systems would add immeasurably to evaluating the risks from those hazards. Using the Department of Energy (DOE) facilities as examples, it is suggested that there are three major types of systems that should be considered: (1) contaminated buildings and other facilities, (2) buffer zones that immediately surround contaminated facilities, and (3) relatively pristine or uncontaminated ecosystems that surround or are adjacent to the contaminated facilities. The relative ecological risks from remediation and restoration on these three different types of systems vary markedly, in terms of immediate effects, long-term effects, and the potential for complete recovery. Remediation on most systems containing predominately buildings and other facilities entails very little ecological risk, except for disruption of nearby ecosystems from construction and transport of materials offsite. Remediation on buffer zones, however, might cause considerable ecological damage if these systems have already undergone partial recovery, are large, and will experience physical disruption during remediation. Remediation that impacts relatively pristine or uncontaminated sites can cause major disruptions to ecosystems well beyond the value, in terms of reduced human health risk, of the restoration itself, and recovery may never be complete. Although this analysis does not consider the risks to workers from remediation, such risks should be added to the potential harm to functioning ecosystems from remediation at nearby contaminated sites. The effects of remediation on all three types of systems should be balanced when making remediation decisions. The newly proposed integrator operable unit, which surrounds the contaminated sites (operable units), and includes all other areas, provides an additional framework for considering risk, remediation, and long-term stewardship on Department of Energy lands. PMID- 11032428 TI - Commentary on plasma levels of homovanillic acid (phVA) under influence of mental stress. PMID- 11032429 TI - Schizophrenia and plasma homovanillic acid in response to mental stress: methodological considerations. PMID- 11032430 TI - Advances in End-Stage Renal Diseases 2000. Proceedings of an international conference on dialysis II. January 13-14, 2000, Tarpon Springs, Florida, USA. PMID- 11032431 TI - Tamoxifen for the prevention of breast cancer. Relationship of trial design to value of data for patient? PMID- 11032432 TI - Carcinoembryonic antigen elevation in agnogenic myeloid metaplasia. PMID- 11032433 TI - Endophthalmitis following globe perforation with a hypodermic needle. PMID- 11032434 TI - Intranasal passage of dacryoliths. PMID- 11032435 TI - Treatment of recurrent giant iris cyst with intracyst administration of mitomycin C. PMID- 11032436 TI - Scleritis as a presenting feature of Takayasu's disease. PMID- 11032437 TI - Acute visual loss by an Onodi cell. PMID- 11032438 TI - Spontaneous arteriovenous malformation of the orbit. PMID- 11032439 TI - Perceptual distortion around homonymous scotomas is not restricted to defects located in the right hemifield. PMID- 11032440 TI - Spontaneous closure of a macular hole secondary to an accidental laser injury. PMID- 11032442 TI - Unusual presentation of diffuse melanoma of the iris. PMID- 11032441 TI - Failure of transpupillary thermotherapy (TTT) for choroidal melanoma: two cases with histopathological correlation. PMID- 11032443 TI - Promoter mutations of myocilin gene in Japanese patients with open angle glaucoma including normal tension glaucoma. PMID- 11032444 TI - Keratoconus associated with chromosome 13 ring abnormality. PMID- 11032445 TI - Vitamin A deficiency in coeliac disease. PMID- 11032446 TI - Choroidal neovascular membrane presenting as a complication of pre-eclampsia in a patient with the antiphospholipid syndrome. PMID- 11032447 TI - Usefulness of three dimensional ultrasonography for invisible fundus. PMID- 11032448 TI - Late complications after pterygium excision with high dose mitomycin C instillation. PMID- 11032450 TI - SCAR 2000. Proceedings of the 17th Symposium for Computer Applications in Radiology. Philadelphia, Pennsylvania, USA. June 3-6, 2000. PMID- 11032449 TI - Department of Health and Human Services. Recombinant DNA Advisory Committee. National Institutes of Health. Minutes of Meeting. June 14, 1999. PMID- 11032451 TI - [Public health of the twenty-first century--future-based environmental action]. PMID- 11032453 TI - First Symposium of the International College of Advanced Longevity Medicine, Reno, Nevada, october 11-13, 1998. PMID- 11032452 TI - Eat to live, not live to eat. AB - Most of the prevailing chronic diseases in the world have an important nutritional component by directly causing a specific disease, enhancing the risk through phenomena of promotion, exerting a beneficial effect in decreasing risk, or preventing the disease. International studies in geographic pathology have shown that a given disease may have vastly different incidence and mortality as a function of residence. Laboratory research in animal models can reproduce fairly accurately what is learned through international research and provide the basis for examining relevant hypotheses and, more importantly, proposed mechanisms of action. Validation of these approaches can be the basis for public-health recommendations and health-promotion activities. Through such techniques, it has been found that regular intake of foods with saturated fats such as meat and certain dairy products raise the risk of coronary heart disease. The total mixed fat intake is associated with a higher incidence of the nutritionally linked cancers, specifically cancer of the postmenopausal breast, distal colon, prostate, pancreas. ovary, and endometrium. The associated genotoxic carcinogens for several of these cancers are heterocyclic amines, which also play a role in heart-disease causation, and these are produced during the broiling and frying of creatinine-containing foods such as meats. Monounsaturated oils such as olive or canola oil are low-risk fats as shown in animal models and through the observation that the incidence of specific diseases is lower in the Mediterranean region, where such oils are customarily used. High salt intake is associated with high blood pressure and with stomach cancer, especially with inadequate intake of potassium from fruits and vegetables and of calcium from certain vegetables and low-fat dairy products. Vegetables, fruits, and soy products are rich in antioxidants that are essential to lower disease risk stemming from reactive oxygen systems in the body. Green and black teas are excellent sources of antioxidants of a polyphenol nature. as is cocoa and some chocolates. Nutritional lifestyles that offer the possibility of a healthy long life can be adopted by most populations in the world. PMID- 11032454 TI - How to provide nutritional support. PMID- 11032455 TI - Underwater and hyperbaric medicine. Abstracts from the literature. PMID- 11032456 TI - Synchrotron radiation in cellular and molecular biology, Part II. Foreword. PMID- 11032457 TI - Health risks from exposure to cadmium in soil. PMID- 11032458 TI - Non-neoplastic mortality of European workers who produce man made mineral fibres. PMID- 11032459 TI - How it was demonstrated that transport of oxygen still requires myoglobin. PMID- 11032460 TI - A direct-method ab initio phasing of a protein, pseudoazurin, at 1.55 A resolution. AB - The direct-method program MULTAN88 has been applied to solve a known protein, pseudoazurin, space group P6(5), unit-cell parameters a = b = 50.0 (1), c = 98.5 (3) A, with 917 protein atoms, a Cu atom and 93 solvent water molecules in the asymmetric unit (>6,000 non-H atoms in the unit cell) and data at 1.55 A resolution. One of several trials with sets of initially random phases yielded phase estimates for 1,000 reflections with a mean phase error of 68.3 degrees that was recognized as the best available solution by the figure of merit being used. Phase extension to 2,000 largest Es showed a distorted tetrahedral geometry around the Cu site. Density modification was applied to improve the phases and the quality of the maps, starting with phases calculated from the atomic positions indicated by the first map. PMID- 11032461 TI - Calreticulin comes of age. PMID- 11032462 TI - Interleukin-1alpha antisense oligomer suppresses hepatocyte growth. PMID- 11032463 TI - Urological Research Society annual scientific meeting. 7 January 2000, London, United Kingdom. Abstracts. PMID- 11032465 TI - Host cell-dependent replication of HIV-1 mutants with deletions in gp41 cytoplasmic tail region is independent of the function of Vif. PMID- 11032464 TI - Amisulpride has a superior benefit/risk profile to haloperidol in schizophrenia: results of a multicentre, double-blind study (the Amisulpride Study Group). AB - In a multicentre, double-blind, flexible-dose study, 199 patients with paranoid schizophrenia or schizophreniform disorders received haloperidol (10-30 mg/d) or amisulpride (400-1200 mg/d) for four months. More patients in the haloperidol group withdrew prematurely (44% vs 26%; P = 0.0077) due to a higher incidence of adverse events. Amisulpride was at least as effective as haloperidol in reducing the Brief Psychiatric Rating Scale (BPRS) total score (-27.3 vs -21.9) (non inferiority test; P < 0.001). The PANSS positive score improved to a similar extent in both groups whilst improvement in the PANSS negative score was significantly greater with amisulpride (-10.5 vs -7.2; P = 0.01). The percentage of responders on the Clinical Global Impression scale was also significantly greater with amisulpride (71% vs 47%; P < 0.001). Both the Quality of Life Scale (QLS) and the Functional Status Questionnaire (FSQ) improved to a significantly greater extent under amisulpride. Haloperidol was associated with a greater incidence in extrapyramidal symptoms and with a greater increase in the Simpson Angus score than was seen with amisulpride (0.32 vs 0.02; P < 0.001). In conclusion, amisulpride is globally superior to haloperidol in the treatment of acute exacerbations of schizophrenia and significantly improves patients' quality of life and social adjustment. PMID- 11032466 TI - Critical Care Issues in Liver Transplantation. Conference proceedings. November 5, 1999. Dallas, Texas, USA. PMID- 11032467 TI - [Laparoscopic treatment of endometriomas: cystectomy or suppression? Against laparoscopic cystectomy]. PMID- 11032468 TI - The generalizability of antiphospholipid testing? PMID- 11032469 TI - Relationship of embryo cryopreservation to cost-effectiveness of ART. PMID- 11032470 TI - The coalescent in an exponentially growing metapopulation and its application to Arabidopsis thaliana. PMID- 11032471 TI - Cemented versus uncemented Thompson's prostheses: a functional outcome study. PMID- 11032472 TI - Cemented versus uncemented Thompson's prostheses: a functional outcome study. PMID- 11032475 TI - Amyloglucosidase hydrolysis of high-pressure and thermally gelatinized corn and wheat starches. AB - The study of glucose production using amyloglucosidase as a biocatalyst was carried out using high-pressure and thermally gelatinized corn and wheat starches. For corn starch, the measured initial rate of glucose production obtained from thermal gelatinization is faster than that obtained from the two high-pressure treatments, but the equilibrium yield of glucose was found to be similar for the three treatments. High-pressure treatments of wheat starch significantly improve the equilibrium yield of glucose compared with those obtained from the thermally gelatinized wheat starch. This difference has been related to the formation of amylose-lipid complexes during heating and could also explain previous physicochemical differences observed between high-pressure and thermally gelatinized starch. PMID- 11032473 TI - The effect of stents on mucosal wound healing. PMID- 11032476 TI - Interactions of different carrageenan isoforms and flour components in breadmaking. AB - The aim of this study was to compare the effects of carrageenans with different sulfate contents on bread volume and dough rheological properties. Results showed that only lambda carrageenan, the most sulfated isoform, produced a significant increase in bread volume. In contrast, the different carrageenans induced a negative effect on the cookie factor. Alveographic and farinographic analyses indicated that dough rheological properties were differentially modified depending on whether lambda carrageenan was added to flour and then hydrated or vice versa. Analysis of the interaction between lambda carrageenan and flour components by infrared spectroscopy and SDS-PAGE indicated that a pool of low molecular weight hydrophobic gluten proteins interact with carrageenan. This interaction drastically changes their physicochemical properties since carrageenan-gluten protein complexes show a hydrophilic behavior. In addition, the results indicate that carrageenan sulfate groups and probably the amino groups of glutamines present in the primary structure of gluten proteins are involved in the interaction. PMID- 11032477 TI - The relationship between high-molecular-weight glutenin subunit composition and the quality of Japanese hexaploid wheat lines. AB - To reveal the high-molecular-weight (1-1MW) glutenin subunit composition, the seed storage proteins of 40 Japanese wheat (Triticum aestivum) lines were fractionated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis to determine their HMW glutenin subunit composition. These were identified by comparison of subunit mobility with that previously found in hexaploid wheat. Twelve different, major glutenin HMW subunits were identified. Each line contained three to five subunits, and 11 different glutenin subunit patterns were observed for 11 alleles in Japanese lines. The Glu-1 quality scores were not particularly high for most of the Japanese wheats in the southern part of Japan (Kyushu district). However, the Glu-1 quality scores of several wheat lines in the Hokkaido area (north Japan) were high. South Japanese wheat lines showed specialty allelic variation in the glutenin HMW 145 kfla subunit, different from those in non-Japanese hexaploid wheats. PMID- 11032478 TI - Content of the flavonols myricetin, quercetin, and kaempferol in finnish berry wines. AB - The amounts of myricetin, quercetin, and kaempferol were analyzed in 16 red and 2 white berry and grape wines after acid hydrolysis using an RP-HPLC method with diode array detection. The red berry wines analyzed were made mainly from black currant, crowberry, and bog whortleberry, i.e., berries rich in flavonols. The red grape wines were made mainly from Cabernet Sauvignon or Merlot grapes in several countries. The white wines studied were gooseberry and white currant wines and Chardonnay and Riesling wines. The amount of myricetin ranged from 3.8 to 22.6 mg L(-1) in red berry wines and from 0 to 14.6 mg L(-1) in red grape wines. The amount of quercetin was from 2.2 to 24.3 mg L(-1) red berry wines and from <1.2 to 19.4 mg L(-1) in red grape wines. Low levels of kaempferol were found in all red berry wines and in 9 red grape wines. The total concentration of these flavonols was from 6 to 46 mg L(-1) (mean 20 mg L(-1)) in red berry wines and from 4 to 31 mg L(-1) (mean 15 mg L(-1)) in red grape wines. Small amounts of quercetin were found in white currant and gooseberry wines, whereas no flavonols were detected in white grape wines. These results demonstrate that the contents of flavonols in red PMID- 11032479 TI - Isolation and characterization of novel stilbene derivatives from Riesling wine. AB - Besides the already known stilbenes trans-resveratrol as well as isomeric piceids seven novel stilbene derivatives have been isolated from a commercial Riesling wine. The newly identified compounds included the monostilbene 2,4,6 trihydroxyphenanthrene-2-O-glucoside, as well as two isomeric resveratrol-2-C glucosides. In addition, four dimeric stilbenes, i.e., cis- and trans-epsilon viniferin diglucoside as well as pallidol glucoside and pallidol diglucoside, have also been obtained for the first time from Riesling wine. PMID- 11032480 TI - Carotenoid composition in the fruits of Asparagus officinalis. AB - The carotenoid pigments of the ripe and unripe fruits of Asparagus officinalis were investigated by means of an HPLC technique. Capsanthin, capsorubin, capsanthin 5,6-epoxide, antheraxanthin, violaxanthin, neoxanthin, mutatoxanthin epimers, zeaxanthin, lutein, beta-cryptoxanthin, beta-carotene, and some cis isomers were found. Carotenoids with 3,5,6-trihydroxy and 3,6-epoxy beta-end groups could not be deleted. PMID- 11032481 TI - Content of phenolic acids and ferulic acid dehydrodimers in 17 rye (Secale cereale L.) varieties. AB - The contents of pnenolic acids and ferulic acid dehydrodimers were quantified by HPLC analysis after alkaline hydrolysis in kernels of 17 rye (Secale cereale L.) varieties grown in one location in Denmark during 1997 and 1998. Significant variations (P < 0.05) with regard to the concentration of the analyzed components were observed among the different rye varieties and also between different harvest years. However, the content of phenolic acids in the analyzed rye varieties was narrow compared to cereals such as wheat and barley. The concentration of ferulic acid, the most abundant phenolic acid ranged from 900 to 1170 microgram g(-1) dry matter. The content in sinapic acid ranged from 70 to 140 microgram g(-1) dry matter, p-coumaric acid ranged from 40 to 70 microgram g( 1) dry matter, and caffeic, p-hydroxybenzoic, protocatechuic, and vanillic acids were all detected in concentrations less than 20 microgram g(-1) dry matter. The most abundant ferulic acid dehydrodimer 8-O-4 -DiFA was quantified in concentrations from 130 to 200 microgram g(-1) dry matter followed by 8,5 -DiFA benzofuran form (50-100 microgram g(-1) dry matter), 5,5 -DiFA (40-70 microgram g(-1) dry matter), and 8,5 -DiFA (20-40 microgram g(-1) dry matter). PMID- 11032482 TI - Use of the MS-sensor to discriminate between different dosages of garlic flavoring in tomato sauce. AB - A method has been developed to discriminate between different dosages of garlic flavoring in tomato sauce with the help of a mass spectrometry based sensory system. Four fragment ions m/z 73, 81, 114, and 120 were selected as "sensor array" during direct injection of the sample headspace into the mass spectrometer. Tomato sauces blended with different types of flavoring could be discriminated, and concentration gradients could be monitored. Fragment ions were chosen after volatile components had been analyzed and identified by SPME-GC/MS and HS-GC/MS (full scan). HS-GC/MS profiles of m/z 73, 81, 114, and 120 were recorded in the selected ion monitoring mode. PMID- 11032483 TI - Effect of high-pressure treatment on lipoxygenase activity. AB - Solutions of commercial soybean lipoxygenase (100 microgram/ML in 0.2 M citrate phosphate and 0.2 M Tris buffer were subjected to pressures of 0.1, 200, 400, and 600 MPa for 20 mm. The enzyme was stable at atmospheric pressure (0.1 MPa) over a wide pH range (5-9). In citrate phosphate buffer, the enzyme had maximum stability over the pH range 58 in untreated samples and after treatment at 200 MPa, but with increasing pressure, the pH stability range become narrower and centered around pH 78. The enzyme was more sensitive to acid than alkali, and at pH 9, it lost virtually all activity after pressurization at 600 MPa for 20 mm in both buffers. The activity of the crude enzyme extracted from tomatoes treated at 200 and 300 MPa for 10 mm was not significantly different from that of the untreated tomatoes, while a pressure of 400 MPa for 10 mm caused a significant decrease in activity and treatment at 600 MPa led to complete and irreversible activity loss. Compared to unpressurized tomatoes, treatment at 600 MPa gave significantly reduced levels of hexanal, cis-3-hexenal, and trans-2-hexenal, which are important contributors to "fresh" tomato flavor, and this was attributed to the inactivation of lipoxygenase. PMID- 11032484 TI - Soybean vegetable protein (Tofu) preserved with high pressure. AB - Tofu is a soybean vegetable protein that Asians have long consumed; its intake is increasing in other countries. Tofu was purchased at a local shop. The tofu samples were already preserved in plastic bags subjected to vacuum and storage (5 C). Tofu samples were subjected to high pressure (HP) of 400 MPa at 5 C for 5, 30, and 45 mm. Microbial analysis, sensorial evaluation, and structure were determined. HP treatment in tofu reduces the microbial population. Most of the microorganisms found in the initial population belonged to the Enterobacteriaceae family, bacteria Gram-negative (no Enterobacteriaceae), and bacteria Gram positive. Alter HP treatment, Hafnia alvei and Bacillus cereus were found. After HP treatment, tofu is a pasteurized product, which is safer in terms of secondary pathogenic microbial contamination. Sensorial test results revealed that treated tofu was acceptable to consumers. The micrographs on the cryofracture observed with a cryoscanning electron microscope revealed a more compact structure after pressure compared with that of untreated samples, but the aggregates in the treated samples were more disperse. PMID- 11032485 TI - Study of the temperature effect on the formation of wheat gluten network: influence on mechanical properties and protein solubility. AB - Modifications of mechanical properties of wheat dough during thermal treatments depend mainly on the capacity of wheat gluten proteins to establish intra- and intermolecular interactions when subjected to high-temperature processing. The present study investigates the effect of thermal treatments on the mechanical properties and protein solubility of wheat gluten-based network. The increase in treatment temperatures (from 80 to 135 C) induces an increase in mechanical resistance of the gluten network (tensile strength increases from 0.26 to 2.04 MPa) and a decrease in deformability (elongation decreases from 468 to 236%). The increase in temperature (from 80 to 135 C) also induces a very strong reduction of protein solubility in 2% SDS (from 68 to 0%) that could be correlated to the mechanical changes observed. It was concluded that the modifications of the wheat gluten network properties seem to depend mainly on the temperature level, as temperatures >108-116 C allow activation of thermosetting reactions. PMID- 11032486 TI - Influence of domestic processing and storage on flavonol contents in berries. AB - Effects of domestic processing and storage on the flavonols quercetin, myricetin, and kaempferol in five berries were studied using an optimized RP-HPLC method with UV and diode array detection after an acid hydrolysis of the corresponding glycosides. In fresh berries, the total content of flavonols was highest in lingonberry (169 mg/kg) and black currant (157 mg/kg), intermediate in bilberry (41 mg/kg) and strawberry (17 mg/kg), and lowest in red raspberry (9.5 mg/kg). Cooking strawberries with sugar to make jam resulted in minor losses (quercetin 15%, kaempferol 18%). During cooking of bilberries with water and sugar to make soup, 40% of quercetin was lost. Traditional preservation of crushed lingonberries in their own juice caused a considerable (40%) loss of quercetin. Only 15% of quercetin and 30% of myricetin present in unprocessed berries were retained in juices made by common domestic methods (steam-extracted black currant juice, unpasteurized lingonberry juice). Cold-pressing was superior to steam extraction in extracting flavonols from black currants. During 9 months of storage at 20 C, quercetin content decreased markedly (40%) in bilberries and lingonberries, but not in black currants or red raspberries. Myricetin and kaempferol were more susceptible than quercetin to losses during storage. PMID- 11032487 TI - Selective enzyme-mediated extraction of capsaicinoids and caratenoids from chili guajillo puya (Capsicum annum L.) using ethanol as solvent. AB - The selective extraction of capsaicinoids and carotenoids from chili guajillo "puya" flour was studied. When ethanol was used as solvent, 80% of capsaicinoids and 73% of carotenoids were extracted, representing an interesting alternative for the substitution of hexane in industrial processes. Additionally, when the flour was pretreated with enzymes that break the cell wall and then dried, extraction in ethanol increased to 11 and 7% for carotenoid and capsaicinoid, respectively. A selective two-stage extraction process after the treatment with enzymes is proposed. The first step uses 30% (v/v) ethanol and releases up to 60% of the initial capsaicinoids, and the second extraction step with industrial ethanol permits the recovery of 83% of carotenoids present in the flour. PMID- 11032488 TI - Platelet SHT and biopolar depressed patients. PMID- 11032489 TI - High opioid costs in Argentina: an availability barrier that can be overcome. PMID- 11032490 TI - Gabapentin, an adjuvant treatment for neuropathic pain in a cancer hospital. PMID- 11032491 TI - [Cause for chronic kidney failure and selection of therapeutic methods]. PMID- 11032492 TI - [Mechanism for the progression of chronic kidney failure]. PMID- 11032493 TI - [Disease stages and physiopathology of chronic kidney failure]. PMID- 11032494 TI - [Treatment of hypertension]. PMID- 11032495 TI - [Chemotherapy to arrest the progression of chronic kidney failure]. PMID- 11032496 TI - [Diet therapy for chronic kidney failure]. PMID- 11032497 TI - [Treatment of diabetic nephropathy]. PMID- 11032498 TI - [Current status of dialysis therapy]. PMID- 11032500 TI - [Home dialysis]. PMID- 11032499 TI - [Introduction of dialysis therapy to patients with chronic renal failure]. PMID- 11032501 TI - [Complications suffered by dialysis patients. 1. Circulatory diseases]. PMID- 11032502 TI - [Complications suffered by dialysis patients. 2. Infections and malignant tumors]. PMID- 11032503 TI - [Complications suffered by dialysis patients. 3. Ca and P metabolism disorders of bone]. PMID- 11032504 TI - [Complications suffered by dialysis patients. 4. Hematologic problems]. PMID- 11032505 TI - [Complications suffered by dialysis patients. 5. Dialysis-induced amyloidosis]. PMID- 11032506 TI - [Problems of aged dialysis patients]. PMID- 11032507 TI - [Problems of diabetic patients under dialysis therapy]. PMID- 11032508 TI - [Practice of kidney transplantation and its complications]. PMID- 11032509 TI - [Therapy of chronic kidney failure--from the conservative stage to transplantation. Discussion]. PMID- 11032511 TI - Parameters of Care. American Academy of Periodontology. PMID- 11032510 TI - [Functional regeneration of pancreatic beta-cells]. PMID- 11032512 TI - The content of rural health research. . PMID- 11032513 TI - JAMA patient page. Depression. PMID- 11032514 TI - Effect of eliminating compensation for pain and suffering on the outcome of insurance claims. PMID- 11032515 TI - Effect of eliminating compensation for pain and suffering on the outcome of insurance claims. PMID- 11032516 TI - Effect of eliminating compensation for pain and suffering on the outcome of insurance claims. PMID- 11032517 TI - Effect of eliminating compensation for pain and suffering on the outcome of insurance claims. PMID- 11032518 TI - Nonaccredited medical education in the United States. PMID- 11032519 TI - Nonaccredited medical education in the United States. PMID- 11032520 TI - Nonaccredited medical education in the United States. PMID- 11032521 TI - Nonaccredited medical education in the United States. PMID- 11032522 TI - Nonaccredited medical education in the United States. PMID- 11032523 TI - Nonaccredited medical education in the United States. PMID- 11032524 TI - Nonaccredited medical education in the United States. PMID- 11032525 TI - Nonaccredited medical education in the United States. PMID- 11032526 TI - Lamivudine in the treatment of acute hepatitis B. PMID- 11032527 TI - Clonal identity of nodular lymphocyte-predominant Hodgkin's disease and T-cell rich B-cell lymphoma. PMID- 11032528 TI - Unapproved and off-label use of drugs in a children's hospital. PMID- 11032529 TI - Airing differences in pediatric nebulizer therapy. PMID- 11032530 TI - Parasitic helminths from genomes to vaccines II: Edinburgh, July 1999. PMID- 11032531 TI - Genetics and nursing: planning for the future. PMID- 11032532 TI - Drug treatment for depression in patients inside rehabilitation wards. PMID- 11032533 TI - Medical urology. PMID- 11032534 TI - Publishing case reports--a powerful tool for academic motivation. PMID- 11032535 TI - Hyperthyroidism in an elderly patient. PMID- 11032536 TI - [The Batista operation: fiction or reality]. PMID- 11032537 TI - [Sequential atrioventricular stimulation in obstructive myocardial hypertrophy. What patients benefit?]. PMID- 11032538 TI - [Criss-cross heart with discordant atrioventricular connection and arterial ventricle]. PMID- 11032539 TI - [Dalteparin in the acute phase of unstable angina and non-Q-wave infarction]. PMID- 11032540 TI - [Safety and economic efficiency of platelet IIb/IIIa receptor blockers in acute coronary syndromes without ST-segment elevation]. PMID- 11032542 TI - [Evidence based medicine]. PMID- 11032541 TI - [The PRIAMHO study and the usefulness of hospital-based registries in acute myocardial infarction]. PMID- 11032543 TI - EEG arousals: scoring rules and examples: a preliminary report from the Sleep Disorders Atlas Task Force of the American Sleep Disorders Association. PMID- 11032544 TI - Induction by soluble factors of organized axial structures in chick epiblasts. AB - Inductive action of soluble factors was tested on isolated chick epiblasts. An assay was developed wherein conditioned medium derived from the Xenopus XTC cell line induced the formation of a full-length notochord and rows of bilaterally symmetric somites. Basic fibroblast growth factor, epidermal growth factor, retinoic acid, and transforming growth factor type B1 and B2 were not capable of inducing axial structures. Thus, soluble factors can elicit the development of polarity stored in the epiblast and behave as true morphogens since they can induce the formation of the organized complex structures that constitute the embryonic axis. PMID- 11032546 TI - NIH. Imaging institute picks up momentum. AB - Last week a House panel approved the creation of the National Institute of Biomedical Imaging and Bioengineering (NIBIE) despite pleas from top National Institutes of Health officials that such a move will balkanize research. The bipartisan proposal, which is also backed by Democratic presidential candidate Al Gore, may have enough momentum to sail through Congress by the time it adjourns early next month. Even if it falls short, NIBIE reflects the growing desire of segments of the biomedical community to have their own address on the Bethesda campus. PMID- 11032545 TI - Viral escape and the failure of cellular immune responses. PMID- 11032547 TI - Virology. Canine virus blamed in Caspian seal deaths. AB - Canine distemper virus has been identified as the most likely cause of a die-off of thousands of seals in the Caspian Sea earlier this year. Although the findings by two independent research groups allay fears of a threat to humans, they heighten concerns about the survival of the imperiled species. PMID- 11032548 TI - Biophysics. For certain shrimp, life's a snap. AB - On page 2114 of this issue, physicists report that a collapsing bubble outside the claw of the snapping shrimp Alpheus heterochaelis causes its characteristic clack. According to this new study, A. heterochaelis clamps its claw so rapidly that a water jet gushing from the claw first loses and then gains pressure, causing an air bubble in the jet to swell and collapse with a pronounced "snap!" The imploding bubble generates shock waves that stun nearby prey and ward off other shrimp, who have learned to keep their distance. PMID- 11032549 TI - Medical ethics. Moratorium urged on germ line gene therapy. PMID- 11032550 TI - Scientific community. Soft money's hard realities. AB - "Second-class citizen" is how researchers on soft money, who have to raise their salaries from grants, describe their position--even those who like their jobs. It can be fraught with financial insecurity, disrespect, and poor facilities--as well as some advantages, such as freedom from administrative duties. Those who have done it say that success requires a strong will, an accommodating department, friends in high places, and money in the bank as a cushion--not to mention emotional security and a tough skin. PMID- 11032551 TI - Molecular biology. New way found to study closely related proteins. AB - Rather than designing specific inhibitors for closely related proteins, researchers are remodeling the proteins to make them uniquely susceptible to inhibition. As described in the 21 September issue of Nature, the technique involves enlarging the active site of an enzyme so that it can bind an inhibitor that won't fit into the active sites of related--but unaltered--enzymes. Researchers can then insert the gene that encodes the modified enzyme into cells or living animals and turn off that enzyme by feeding them the inhibitor--without affecting other, very similar, enzymes. The technique may have some advantages over other approaches to studying the functions of individual proteins, such as mutating or knocking out the genes that encode them, which may disrupt embryonic development, producing abnormal animals or no animals at all. PMID- 11032552 TI - Ecological Society of America meeting. Global warming, insects take the stage at Snowbird. AB - Despite the turmoil of not-so-distant forest fires and United Airlines troubles that threw off travel schedules, some 2600 ecologists made their way to this sun soaked canyon last month for the Ecological Society of America's 85th annual meeting. Topics ranged from ancient droughts to photosynthesis beneath snow and how trees resist insects. PMID- 11032553 TI - China's new forest policy. PMID- 11032554 TI - Disaster management. U.S. policies pertaining to weather and climate extremes. AB - Climate and weather extremes--floods, drought, storms, severe weather--now have an impact on all levels of government and on the insurance industry. U.S. national policy has moved from structural approaches (building dams, levees, and irrigation ditches) to societal approaches: moving people out of hazardous areas, seeking improvements in building codes, and encouraging the use of crop and flood insurance. Federal relief assistance policies have been called into question, and renewed emphasis on personal responsibility is part of future policy. PMID- 11032555 TI - Climate change. Managing forests after Kyoto. AB - The Kyoto protocol aims to reduce carbon emissions into the atmosphere. Part of the strategy is the active management of terrestrial carbon sinks, principally through afforestation and reforestation. In their Perspective, Schulze et al. argue that the preservation of old-growth forests may have a larger positive effect on the carbon cycle than promotion of regrowth. PMID- 11032556 TI - Biochemistry. Ubiquitination--more than two to tango. AB - The ubiquitin pathway in the cell is an elegant system for targeting unwanted proteins for degradation. Three enzymes, E1, E2, and E3, are responsible for attaching the ubiquitin tag to proteins destined to be chopped up. In their Perspective, Joazeiro and Hunter discuss new structural findings that reveal the part played by an E3 called c-Cbl in this ubiquitinating process. PMID- 11032557 TI - Aging, chromatin, and food restriction--connecting the dots. AB - Model organisms such as yeast have proved exceptionally valuable for revealing new information about the molecular pathways involved in the aging of cells. In her Perspective, Campisi comments on new work showing that caloric restriction increases longevity in yeast by activating the SIR2 gene, which alters the compactness of chromatin and thus regulates gene expression (Lin et al.). PMID- 11032558 TI - Biomedicine. Staying slim with insulin in mind. AB - Striking the delicate balance between energy intake in the form of food and energy expenditure in the form of metabolic activity keeps the body extremely busy. As Schwartz explains in his enlightening Perspective, the finding that insulin signals the brain to promote weight loss (Bruning et al.) flies in the face of the notion that insulin is involved solely in glucose storage, its conversion to fat, and weight gain. PMID- 11032559 TI - [What will internal medicine look like in the next century in the Czech Republic]. PMID- 11032560 TI - [The more specialists, the worse the results? Comment on the article by Dr. P. Widimsky "What will internal medicine look like in developed countries in the 21st century?". PMID- 11032561 TI - [The concept of internal medicine as a profession]. PMID- 11032562 TI - What is a safe vaccine? PMID- 11032563 TI - Evaluation of the potency of BIKEN inactivated Japanese Encephalitis vaccine and DNA vaccines in an intracerebral Japanese Encephalitis virus challenge. PMID- 11032564 TI - [Prof. I. I. Rogozin (1900-1973)]. PMID- 11032565 TI - [The 70th anniversary of the Nizhegorod Research Institute of Epidemiology and Microbiology of the Ministry of Public Health of the Russian Federation]. PMID- 11032566 TI - Nonalcoholic fatty liver disease is associated with hyperlipidemia and obesity. PMID- 11032567 TI - Basic biomedical science and the destruction of the pathophysiologic bridge from bench to bedside. PMID- 11032568 TI - Is screening mammography effective in elderly women. PMID- 11032569 TI - Nephrotic-range proteinuria in patients with renovascular disease. PMID- 11032570 TI - Primary amyloidosis presenting as dilated veins. PMID- 11032571 TI - Neisseria sicca meningitis in a woman with nascent pernicious anemia. PMID- 11032572 TI - Hepatitis C virus infection manifesting as tubulointerstitial nephritis, cardiomyopathy, and hepatitis. PMID- 11032573 TI - TKY101: a highly polymorphic equine dinucleotide repeat locus. PMID- 11032574 TI - [Bibliography of the scientific publications of the Pomeranian Medical Academy in Szczecin for 1997]. PMID- 11032575 TI - [Medical Society of Paris Hospitals]. PMID- 11032576 TI - Cystic fibrosis related low bone density. PMID- 11032577 TI - Non-familial short stature. PMID- 11032578 TI - Guidelines for the ethical conduct of medical research involving children. PMID- 11032579 TI - Guidelines for the ethical conduct of medical research involving children. PMID- 11032580 TI - Physical treatment of fever. PMID- 11032581 TI - The role of lumbar puncture in meningococcal disease. PMID- 11032582 TI - A short stay observation unit improves care in the paediatric emergency care setting. PMID- 11032583 TI - Health care refugees. PMID- 11032584 TI - Gynecologic surveillance of women on tamoxifen: first do no harm. PMID- 11032585 TI - Effect of adjuvant tamoxifen on the endometrium in women with breast cancer: a prospective study using office endometrial biopsy. AB - PURPOSE: To determine the frequency of developing abnormal pathologic changes in the endometria of tamoxifen-treated women. To characterize the type of pathologic changes involved. PATIENTS AND METHODS: Between October 1991 and September 1998, 159 patients initiating tamoxifen therapy for breast cancer confined to the breast and axillary lymph nodes were entered in a prospective study. In this study, office endometrial biopsies (EMBs) were obtained during the initiation of tamoxifen and at 6-month intervals for a 2-year period. Three subsequent annual EMBs were recorded for each patient, amounting to a 5-year surveillance. RESULTS: One hundred fifty-nine patients with a median age of 50 years were entered onto study. Patients were assessable if EMBs were performed at least 1 year after the initiation of tamoxifen treatment. Nine patients (5. 7%) were considered protocol violations. The remaining 111 assessable patients underwent a total of 635 EMBs (mean, 5.8 EMBs), with a median surveillance time of 36 months. Eighty-two (12.9%) of the 635 biopsies revealed tissue insufficient for diagnosis. Fourteen patients (12.6%) underwent dilation and curettage (D&C) for an abnormal EMB, persistent bleeding, or for evaluation of adnexal masses at the time of laparoscopy. Findings at D&C included complex hyperplasia (n = 1), abnormal histiocytes (n = 1), simple hyperplasia (n = 2), polyps (n = 4), endocervical polyp (n = 1), and decidualization (n = 2). Three D&Cs were negative. Three patients have undergone hysterectomy. CONCLUSION: EMB was used to monitor the endometrium in the majority (95%) of breast cancer patients on tamoxifen in this trial, but the utility of routine EMB for screening in tamoxifen-treated women seems limited. PMID- 11032586 TI - Effects of adjuvant tamoxifen on the endometrium in postmenopausal women with breast cancer: a prospective long-term study using transvaginal ultrasound. AB - PURPOSE: To study the value of transvaginal ultrasound (TVS) in endometrial screening of postmenopausal breast cancer patients treated with tamoxifen. PATIENTS AND METHODS: In 247 tamoxifen-treated (20 to 30 mg/d for >/= 2 years) women and 98 controls, the endometrium was prospectively followed-up by means of TVS every 6 months for up to 5 years. Patients with homogeneous endometrium of more than 10-mm thickness were then scanned repeatedly every 3 months. RESULTS: The mean endometrial thickness was 3.5 +/- 1.1 mm before treatment and increased to a maximum of 9. 2 +/- 5.1 mm after 3 years of tamoxifen application (P: <.0001), which was significantly (P: <.0001) thicker compared with controls. Fifty-two asymptomatic patients with thickened or morphologically suspect endometrium underwent hysteroscopy and dilatation and curettage (D&C), resulting in four uterine perforations. Histopathologically, atrophy was found in 38 patients (73.1%), polyps in nine, hyperplasia in four, and endometrial cancer in one case. In 20 screened patients who reported vaginal bleeding, five atrophies (25%), five polyps, four hyperplasias, and two endometrial cancers were found. Before hysteroscopy and D&C were performed, 36 (69.2%) of 52 asymptomatic and four (20%) of 20 symptomatic patients were scanned by repeated TVS over 2 to 30 months. Invasive diagnostic procedures were significantly (P: <.05) more frequent in younger and obese patients. In the controls, one asymptomatic polyp and one symptomatic hyperplasia were found. CONCLUSION: In tamoxifen-treated patients, TVS offered a high false-positive rate, even with a cutoff value of 10 mm for endometrial thickness and repeated TVS scans. Increased iatrogenic morbidity and only one asymptomatic endometrial carcinoma do not warrant endometrial screening by TVS in tamoxifen-treated patients. PMID- 11032587 TI - HER-2/neu and p53 expression versus tamoxifen resistance in estrogen receptor positive, node-positive breast cancer. AB - PURPOSE: An association between the overexpression of proto-oncogene HER-2/neu and resistance to tamoxifen in estrogen receptor (ER)-positive primary and metastatic breast cancer has been suggested. We examine a possible interaction between HER-2/neu or p53 expression and tamoxifen effectiveness in patients with ER-positive, node-positive disease treated with cyclophosphamide, doxorubicin, and fluorouracil in a large adjuvant chemotherapy trial (Cancer and Leukemia Group B [CALGB] 8541). Tamoxifen assignment was not randomized-physician discretion was used for premenopausal and postmenopausal women. Trial protocol then specified assignment to postmenopausal women with ER-positive tumors, although not all took tamoxifen. PATIENTS AND METHODS: CALGB 8541 assessed HER 2/neu expression in patients with ER-positive disease by immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) and amplification by differential polymerase chain reaction (PCR). IHC assessed expression of p53. Univariate and multivariate proportional hazards models assessed tamoxifen-HER 2/neu status interactions and tamoxifen-p53 status interactions. RESULTS: HER 2/neu status was available for 651 patients with ER-positive disease; 650, 608, and 353 patients were assessed by IHC, PCR, and FISH, respectively. Approximately one half received tamoxifen. Reduction in risk of disease recurrence or death resulting from tamoxifen was approximately 37% (32% with overexpression and 39% with normal expression of HER-2/neu; n = 155 by IHC). The tamoxifen-HER-2/neu status interaction was not significant in multivariate analysis of all three HER 2/neu assessment methods. Tamoxifen-p53 interaction did not significantly predict outcome. CONCLUSION: Disease-free and overall survival benefit of tamoxifen in patients with ER-positive, node-positive breast cancer does not depend on HER 2/neu or p53 status. Our data suggest that neither HER-2/neu nor p53 expression should be used to determine assignment of tamoxifen. PMID- 11032588 TI - Sentinel lymph node biopsy is accurate after neoadjuvant chemotherapy for breast cancer. AB - PURPOSE: Sentinel lymph node (SLN) biopsy has proved to be an accurate method for detecting nodal micrometastases in previously untreated patients with early-stage breast cancer. We investigated the accuracy of this technique for patients with more advanced breast cancer after neoadjuvant chemotherapy. PATIENTS AND METHODS: Patients with stage II or III breast cancer who had undergone doxorubicin-based neoadjuvant chemotherapy before breast surgery were eligible. Intraoperative lymphatic mapping was performed with peritumoral injections of blue dye alone or in combination with technetium-labeled sulfur colloid. All patients were offered axillary lymph node dissection. Negative sentinel and axillary nodes were subjected to additional processing with serial step sectioning and immunohistochemical staining with an anticytokeratin antibody to detect micrometastases. RESULTS: Fifty-one patients underwent SLN biopsy after neoadjuvant chemotherapy from 1994 to 1999. The SLN identification rate improved from 64.7% to 94.1%. Twenty-two (51.2%) of the 43 successfully mapped patients had positive SLNs, and in 10 of those 22 patients (45.5%), the SLN was the only positive node. Three patients had false-negative SLN biopsy; that is, the sentinel node was negative, but at least one nonsentinel node contained metastases. Additional processing revealed occult micrometastases in four patients (three in sentinel nodes and one in a nonsentinel node). CONCLUSION: SLN biopsy is accurate after neoadjuvant chemotherapy. The SLN identification improved with experience. False-negative findings occurred at a low rate throughout the series. This technique is a potential way to guide the axillary treatment of patients who are clinically node negative after neoadjuvant chemotherapy. PMID- 11032589 TI - Safety and efficacy results of a randomized trial comparing adjuvant toremifene and tamoxifen in postmenopausal patients with node-positive breast cancer. Finnish Breast Cancer Group. AB - PURPOSE: In this multicenter trial, toremifene 40 mg/d was compared with tamoxifen 20 mg/d, both given orally for 3 years to postmenopausal, axillary node positive women after breast surgery. PATIENTS AND METHODS: The first 899 patients (toremifene, n = 459; tamoxifen, n = 440) of the total of 1,480 patients accrued to the trial were included in this scheduled safety analysis. The mean follow-up time was 3.4 years. RESULTS: The two treatment groups were well balanced with respect to patient and disease characteristics. The subjective side-effect profile was similar in both treatment groups. Slightly more vascular complications (deep vein thromboses, cerebrovascular events, and pulmonary embolisms) were seen among tamoxifen-treated patients (5.9%) as compared with toremifene-treated patients (3.5%) (P: =.11), whereas bone fractures (P: =.09) and vaginal leukorrhea (P: =.05) were more common in the toremifene group. The number of subsequent second cancers was similar. The breast cancer recurrence rate was 23.1% (n = 106) in the toremifene group and 26.1% (n = 115) in the tamoxifen group (P: =.31). When only patients with estrogen receptor (ER) positive cancer were considered (n = 556), the risk for breast cancer recurrence was nonsignificantly lower among the toremifene-treated women, with a hazards ratio of 0.74 (90% confidence interval, 0.52 to 1.04; P: =.14). The mean time to breast cancer recurrence and overall survival were similar in both groups. CONCLUSION: The side-effect profile of toremifene resembles that of tamoxifen. The efficacy of toremifene seems to be no less than that of tamoxifen. The trend for fewer breast cancer recurrences in the ER-positive subgroup is encouraging, but a longer follow-up is needed to confirm this. PMID- 11032590 TI - Breast imaging with positron emission tomography and fluorine-18 fluorodeoxyglucose: use and limitations. AB - PURPOSE: To evaluate the diagnostic value of positron emission tomography (PET) using fluorine-18 fluorodeoxyglucose (FDG) for the diagnosis of primary breast cancer. PATIENTS AND METHODS: Preoperatively, 144 patients with masses suggestive of breast cancer underwent PET imaging of the breast. To identify breast cancer by increased metabolic activity, parametric FDG-PET images were analyzed for increased tracer uptake applying conventional image reading (CIR) and sensitive image reading (SIR). One hundred eighty-five breast tumors were evaluated by histology, revealing 132 breast carcinomas and 53 benign masses. RESULTS: Breast carcinomas were identified with an overall sensitivity of 64.4% (CIR) and 80.3% (SIR). The increase in sensitivity (SIR) resulted in a noticeable decrease in specificity, from 94.3% (CIR) to 75.5% (SIR). At stage pT1, only 30 (68.2%) of 44 breast carcinomas were detected, compared with 57 (91.9%) of 62 at stage pT2. A higher percentage of invasive lobular carcinomas were false-negative (65.2%) compared with invasive ductal carcinomas (23.7%). Nevertheless, positive PET scans provided a high positive-predictive value (96.6%) for breast cancer. CONCLUSION: Partial volume effects and varying metabolic activity (dependent on tumor type) seem to represent the most significant limitations for the routine diagnostic application of PET. The number of invasive procedures is therefore unlikely to be significantly reduced by PET imaging in patients presenting with abnormal mammography. However, the high positive-predictive value, resulting from the increased metabolic activity of malignant tissue, may be used with carefully selected subsets of patients as well as to determine the extent of disease or to assess therapy response. PMID- 11032591 TI - Routine chest roentgenography is unnecessary in the work-up of stage I and II breast cancer. AB - PURPOSE: Clinical practice guidelines of many professional societies call for routine staging chest x-rays (SCXR) for all patients with invasive cancer. Given the estimated 157,000 patients annually for whom this recommendation pertains, this screening examination represents a considerable health care expenditure. If it were shown that SCXR rarely changed the management of low-risk subsets of this population, it might be possible to selectively omit this practice from the care of these patients with substantial resultant cost savings. PATIENTS AND METHODS: All patients with clinical stage I and II breast cancer presenting to the Baystate Medical Center from 1989 through 1997 were identified through the Tumor Registry. Their hospital records were reviewed for clinical presentation and documentation of SCXR. RESULTS: One thousand four hundred ninety-four patients were identified with clinical stage I and II disease. SCXR were available for review on 1,003 patients. Only one asymptomatic patient was upstaged to stage IV based on a SCXR. Two patients with primary lung tumors were also identified. These data demonstrate an asymptomatic pulmonary metastasis detection rate of 0. 099% (95% confidence interval, 0.0% to 0.6%). The total charges of SCXR for this group approached $180,000. CONCLUSION: These data demonstrate the low diagnostic yield and high cost of routine SCXR in the management of asymptomatic patients with clinical stage I and stage II breast cancer. Because other studies have shown that SCXR changes neither quality of life nor overall survival, SCXR should be limited to symptomatic patients in whom metastatic disease is suspected. PMID- 11032592 TI - Tamoxifen adjuvant treatment duration in early breast cancer: initial results of a randomized study comparing short-term treatment with long-term treatment. Federation Nationale des Centres de Lutte Contre le Cancer Breast Group. AB - PURPOSE: In 1986, The Federation Nationale desCentres de Lutte Contre le Cancer Breast Group initiated a multicenter randomized trial to assess the usefulness of long-term adjuvant tamoxifen treatment. Short-term adjuvant tamoxifen treatment was to be compared with life long adjuvant tamoxifen treatment. PATIENTS AND METHODS: Patients who were disease-free after 2 to 3 years of adjuvant tamoxifen treatment were eligible for the trial. From September 1986 to May 1995, 3,793 patients were randomized from France, Belgium, and Argentina. A total of 1,882 patients stopped tamoxifen (short-term group), and 1,911 patients were to continue tamoxifen for life (long-term group) at the same dose as previously prescribed. The protocol was modified in February 1997, limiting tamoxifen treatment to 10 years after randomization, thus giving a comparison between a 2- to 3-year treatment and a 12- to 13-year treatment. To date, the median duration of tamoxifen treatment is 30 months in the short-term group, and 70 months in the long-term group. RESULTS: Overall, longer tamoxifen treatment induced a 23% reduction in relapse rates, leading to a 7-year disease-free survival rate of 78%, compared with 72% in the shorter-treatment group. In contrast, overall survival did not differ between the two groups, with a 79% overall survival rate in both groups. This improvement in disease-free survival could be observed in node-positive patients (P: =.001); however, it was not found in node-negative patients. Prolonged tamoxifen treatment corresponded to a significant increase in disease-free survival in estrogen receptor-positive patients (P: =.03) as well as in estrogen receptor-negative patients (P: =.05). Furthermore, longer treatment reduced contralateral breast cancers and did not increase the number of endometrial cancers. CONCLUSION: Although no survival advantage was noted, patients did benefit from longer tamoxifen treatment over 3 years and had significantly better disease-free survival compared with patients who stopped hormonal treatment. Long-term follow-up is needed to assess these results. Most patients in the long-term group are still receiving treatment. Comparison of results as time passes will enable conclusions to be made on the value of long term treatment over 5 years compared with 2 to 3 years. PMID- 11032593 TI - Homoharringtonine and low-dose cytarabine in the management of late chronic-phase chronic myelogenous leukemia. AB - PURPOSE: : To evaluate the efficacy and toxicity profiles of a combination regimen of homoharringtonine (HHT) and low-dose cytarabine (ara-C) in patients with Philadelphia chromosome (Ph)-positive chronic myelogenous leukemia (CML) who had experienced treatment failure with interferon alfa (IFNalpha) therapy. PATIENTS AND METHODS: One hundred five patients were treated: 100 in chronic phase (15 with cytogenetic clonal evolution) and five in accelerated phase. Their median age was 52 years; all had been treated unsuccessfully with IFNalpha; 94% were in late chronic phase; 43% had been exposed to ara-C and 11% had been exposed to HHT. Patients received HHT 2.5 mg/m(2) by continuous infusion daily for 5 days and ara-C 15mg/m(2) daily in two subcutaneous injections for 5 days every 4 weeks. The outcome of the 100 patients in chronic phase was compared with a previous study group of 73 patients treated with HHT alone. RESULTS: Overall, the complete hematologic response (CHR) rate in chronic phase was 72%; the cytogenetic response rate was 32% (major response, 15%; complete response, 5%). Toxicities were acceptable, mostly related to moderate diarrhea (3%), headaches (3%), cardiovascular events (3%),and myelosuppression-associated complications (3% to 14%). With a median follow-up period of 25 months, the estimated 4-year survival rate was 55%. Response rates were identical with HHT plus ara-C versus HHT alone, but the survival was significantly longer with the combination after accounting for differences in the study groups and by multivariate analysis. CONCLUSION: The combination regimen of HHT and ara-C is effective and safe in patients with CML who have experienced treatment failure with IFNalpha and needs to be investigated together with IFNalpha as part of front-line CML therapy. The addition of ara-C did not improve the response rates but may have improved survival, perhaps through suppression of clones related to disease transformation. PMID- 11032594 TI - Phase I trial of carmustine plus O6-benzylguanine for patients with recurrent or progressive malignant glioma. AB - PURPOSE: The major mechanism of resistance to alkylnitrosourea therapy involves the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT), which removes chloroethylation or methylation damage from the O(6) position of guanine. O(6)-benzylguanine (O(6)-BG) is an AGT substrate that inhibits AGT by suicide inactivation. We conducted a phase I trial of carmustine (BCNU) plus O(6)-BG to define the toxicity and maximum-tolerated dose (MTD) of BCNU in conjunction with the preadministration of O(6)-BG with recurrent or progressive malignant glioma. PATIENTS AND METHODS: Patients were treated with O(6)-BG at a dose of 100 mg/m(2) followed 1 hour later by BCNU. Cohorts of three to six patients were treated with escalating doses of BCNU, and patients were observed for at least 6 weeks before being considered assessable for toxicity. Plasma samples were collected and analyzed for O(6)-BG, 8-oxo-O(6)-BG, and 8-oxoguanine concentration. RESULTS: Twenty-three patients were treated (22 with glioblastoma multiforme and one with anaplastic astrocytoma). Four dose levels of BCNU (13.5, 27, 40, and 55 mg/m(2)) were evaluated, with the highest dose level being complicated by grade 3 or 4 thrombocytopenia and neutropenia. O(6)-BG rapidly disappeared from plasma (elimination half-life = 0. 54 +/- 0.14 hours) and was converted to a longer lived metabolite, 8-oxo-O(6)-BG (elimination half-life = 5.6 +/- 2.7 hours) and further to 8-oxoguanine. There was no detectable O(6)-BG 5 hours after the start of the O(6)-BG infusion; however, 8-oxo-O(6)-BG and 8-oxoguanine concentrations were detected 25 hours after O(6)-BG infusion. The mean area under the concentration-time curve (AUC) of 8-oxo-O(6)-BG was 17.5 times greater than the mean AUC for O(6)-BG. CONCLUSION: These results indicate that the MTD of BCNU when given in combination with O(6)-BG at a dose of 100 mg/m(2) is 40 mg/m(2) administered at 6-week intervals. This study provides the foundation for a phase II trial of O(6)-BG plus BCNU in nitrosourea-resistant malignant glioma. PMID- 11032595 TI - Phase I study of preoperative oral uracil and tegafur plus leucovorin and radiation therapy in rectal cancer. AB - PURPOSE: Preoperative combined-modality therapy for rectal cancer may allow for sphincter preservation, while decreasing recurrence rates and improving the overall prognosis. Oral chemotherapy with uracil and tegafur (UFT) plus leucovorin (LV) may reduce costs and complications associated with protracted infusions of fluorouracil. Our goal was to evaluate the safety of UFT plus LV combined with preoperative radiation and determine the maximum-tolerated dose (MTD) and dose-limiting toxicity (DLT) of UFT plus LV in this setting. PATIENTS AND METHODS: Patients with tumor-node-metastasis stage II or III rectal cancer received escalating doses of UFT (starting at 250mg/m(2)/d, with 50-mg/m(2)/d increments between consecutive cohorts) and fixed doses of LV (90 mg/d). The UFT and LV combination was given 5 days per week concurrently with a 5-week course of preoperative radiation totaling 45 Gy (1.8 Gy/fraction). Surgery was performed 4 to 6 weeks after radiation and was followed by four 35-day cycles of fixed doses of UFT and LV (28 days of therapy each cycle). RESULTS: Fifteen patients were treated, and 13 received the full preoperative chemotherapy. All planned radiation was delivered successfully. The MTD of UFT with radiation was 350 mg/m(2)/d with 90 mg/d of LV. Diarrhea was the DLT. Sphincter-preserving surgery was performed in 12 of 14 patients. One patient had progressive disease before surgery. Pathologic evaluation of 14 resected specimens showed a complete response in three cases. CONCLUSION: Preoperative chemoradiation with oral UFT plus LV is feasible and well tolerated and should be further investigated. PMID- 11032596 TI - Phase I trial of 6-hour infusion of glufosfamide, a new alkylating agent with potentially enhanced selectivity for tumors that overexpress transmembrane glucose transporters: a study of the European Organization for Research and Treatment of Cancer Early Clinical Studies Group. AB - PURPOSE: To determine the maximum-tolerated dose (MTD), the principal toxicities, and the pharmacokinetics of 6-hour infusion of glufosfamide (beta-D glucosylisophosphoramide mustard; D-19575), a novel alkylating agent with the potential to target the glucose transporter system. PATIENTS AND METHODS: Twenty one patients (10 women and 11 men; median age, 56 years) with refractory solid tumors were treated with doses ranging from 800 to 6,000 mg/m(2). Glufosfamide was administered every 3 weeks as a two-step (fast/slow) intravenous infusion over a 6-hour period. All patients underwent pharmacokinetic sampling at the first course. RESULTS: The MTD was 6,000 mg/m(2). At this dose, two of six patients developed a reversible, dose-limiting renal tubular acidosis and a slight increase in serum creatinine the week after the second and third courses of treatment, respectively, whereas three of six patients experienced short-lived grade 4 neutropenia/leukopenia. Other side effects were generally mild. Pharmacokinetics indicated linearity of area under the time-versus-concentration curve against dose over the dose range studied and a short elimination half-life. There was clear evidence of antitumor activity, with a long-lasting complete response of an advanced pancreatic adenocarcinoma and minor tumor shrinkage of two refractory colon carcinomas and one heavily pretreated breast cancer. CONCLUSION: The principal toxicity of 6-hour infusion of glufosfamide is reversible renal tubular acidosis, the MTD is 6,000 mg/m(2), and the recommended phase II dose is 4, 500 mg/m(2). Close monitoring of serum potassium and creatinine levels is suggested for patients receiving glufosfamide for early detection of possible renal toxicity. Evidence of antitumor activity in resistant carcinomas warrants further clinical exploration of glufosfamide in phase II studies. PMID- 11032597 TI - Phase I and pharmacokinetic study of docetaxel and irinotecan in patients with advanced solid tumors. AB - PURPOSE: We conducted a phase I and pharmacokinetic study of docetaxel in combination with irinotecan to determine the dose-limiting toxicity (DLT), the maximum-tolerated dose (MTD), and the dose at which at least 50% of the patients experienced a DLT during the first cycle, and to evaluate the safety and pharmacokinetic profiles in patients with advanced solid tumors. PATIENTS AND METHODS: Patients with only one prior chemotherapy treatment (without taxanes or topoisomerase I inhibitors) for advanced disease were included in the study. Docetaxel was administered as a 1-hour IV infusion after premedication with corticosteroids followed immediately by irinotecan as a 90-minute IV infusion, every 3 weeks. No hematologic growth factors were allowed. RESULTS: Forty patients were entered through the following seven dose levels (docetaxel/irinotecan): 40/140 mg/m(2), 50/175 mg/m(2), 60/210 mg/m(2), 60/250 mg/m(2), 60/275 mg/m(2), 60/300 mg/m(2), and 70/250 mg/m(2). Two hundred cycles were administered. Two MTDs were determined, 70/250 mg/m(2) and 60/300 mg/m(2); the DLTs were febrile neutropenia and diarrhea. Neutropenia was the main hematologic toxicity, with 85% of patients experiencing grade 4 neutropenia. Grade 3/4 nonhematologic toxicities in patients included late diarrhea (7.5%), asthenia (15.0%), febrile neutropenia (22.5%), infection (7.5%), and nausea (5.0%). Pharmacokinetics of both docetaxel and irinotecan were not modified with the administration schedule of this study. CONCLUSION: The recommended dose of docetaxel in combination with irinotecan is 60/275 mg/m(2), respectively. At this dose level, the safety profile is manageable. The activity of this combination should be evaluated in phase II studies in different tumor types. PMID- 11032598 TI - Phase I chemotherapy study of biochemical modulation of folinic acid and fluorouracil by gemcitabine in patients with solid tumor malignancies. AB - PURPOSE: This phase I biochemical modulation study evaluated the maximum tolerated dose (MTD), toxicity, and effectiveness of the combination of folinic acid (FA)/fluorouracil (5-FU) followed by escalated dose levels of gemcitabine (FFG) in patients with advanced solid tumors. PATIENTS AND METHODS: Patients were refractory to primary treatment and/or without effective treatment options. Twenty-eight patients received an intravenous (IV) infusion of FA 100 mg/m(2) over 1 hour and a 5-FU 450 mg/m(2) IV bolus in the middle of the FA infusion. After the FA infusion, gemcitabine was administered at a steady rate of infusion of 10 mg/m(2)/min over initially 30 minutes and with increases of an additional 15 minutes at each given level. One cycle consisted of six weekly treatments followed by a 2-week rest. RESULTS: The MTD of gemcitabine was established at 900 mg/m(2) given over 90 minutes. Eight patients of 21 with metastatic colorectal cancer achieved responses (one complete response; seven partial responses), for a response rate of 38%. Responses were seen across the gemcitabine doses of 300 to 900 mg/m(2). One patient had prior treatment with FA/5-FU for advanced disease. Patients with colorectal carcinoma had a median survival of 18 months, and the patient with lung carcinoma has been alive for 24+ months. CONCLUSION: The combination chemotherapy of FFG was well tolerated and may benefit patients with advanced colorectal carcinoma. A phase II evaluation in this patient population is in progress. PMID- 11032599 TI - 2000 update of recommendations for the use of hematopoietic colony-stimulating factors: evidence-based, clinical practice guidelines. American Society of Clinical Oncology Growth Factors Expert Panel. PMID- 11032600 TI - 2000 update of American Society of Clinical Oncology colorectal cancer surveillance guidelines. PMID- 11032601 TI - Errata PMID- 11032602 TI - Progressive multifocal leucoencephalopathy: progress in the AIDS era. PMID- 11032603 TI - Visual rating and volumetry of the medial temporal lobe on magnetic resonance imaging in dementia. PMID- 11032604 TI - Aneurysmal haemorrhage: partialling out the effect of the bleed. PMID- 11032606 TI - Henry Head (1861-1940). PMID- 11032605 TI - Psychological impact of genetic testing for Huntington's disease: an update of the literature. AB - Genetic testing has been available for Huntington's disease for longer than any other adult onset genetic disorder. The discovery of the genetic mutation causing Huntington's disease made possible the use of predictive testing to identify currently unaffected carriers. Concerns have been raised that predictive testing may lead to an increase in deaths by suicide among identified carriers, and these concerns set in motion research to assess the psychological impact of predictive testing for Huntington's disease. This review article provides an overview of the literature and draws implications for clinical practice. About 10%-20% of people at risk request testing when approached by registries or testing centres. Most of the evidence suggests that non-carriers and carriers differ significantly in terms of short term, but not long term, general psychological distress. Adjustment to results was found to depend more on psychological adjustment before testing than the testing result itself. Although risk factors for psychological sequelae have been identified, few adverse events have been described and no obvious contraindications for testing people at risk have been identified. The psychological impact of testing may depend on whether testing was based on linkage analysis or mutation detection. Cohorts enrolled in mutation detection programmes have higher levels of depression before and after testing, compared with people who sought genetic testing when linkage analysis was available. There is evidence that people who choose to be tested are psychologically selected for a favourable response to testing. The impact of testing on people in settings where less intensive counselling protocols and eligibility criteria are used is unknown, and genetic testing is therefore best offered as part of comprehensive specialist counselling. PMID- 11032607 TI - Paradox of a better test for Huntington's disease. AB - OBJECTIVES: To describe the consequences of the identification of the Huntington's disease (HD) mutation on predictive and prenatal testing. METHODS: A retrospective study was performed considering the test applicants, procedures, and results before and after the identification of the mutation. 1032 people at risk for Huntington's disease in The Netherlands were included, of whom 741 applied for the predictive test in the period 1987 to 1997 in Leiden at the Department of Clinical Genetics, and after 1994, also in the other seven clinical genetics departments in The Netherlands. Uptake, sociodemographic variables, and test results, taken before and after the mutation was identified, are described. RESULTS: The uptake of the predictive test in the period studied was 24% and for the prenatal test 2%. No differences were noted in numbers and sociodemographic data between the period before and after the mutation was identified. After an initial increase in test applicants, a decrease was seen after 1995. After 1993 a significant increase of 25% at risk test applicants and a significant decrease of prenatal exclusion tests was noticed. Only 7% asked for reassessment by mutation analysis. New problems arose after the identification of the mutation, such as the option of reassessing the risk obtained by linkage analysis, direct mutation testing of 25% at risk persons with a parent who does not wish to know, new choices regarding reproduction, and new uncertainties for carriers of intermediate and reduced penetrance alleles and for their offspring and relatives. CONCLUSIONS: Although predictive testing has become reliable and available for every person at risk since the mutation has been identified, the uptake of predictive and prenatal tests fell short of expectation, no change in sociodemographic variables was seen, and a decrease in number of applicants was noted. Furthermore, new uncertainties, psychological problems, and questions arose. PMID- 11032608 TI - Health related quality of life in Parkinson's disease: a prospective longitudinal study. AB - OBJECTIVES: To examine the change over time in health related quality of life (HRQL) in a community based cohort of patients with Parkinson's disease. METHODS: One hundred and eleven patients were evaluated for HRQL in 1993 and then again in a follow up study 4 years later. The patients included in the study in 1993 were derived from a prevalence study of patients with Parkinson's disease in the county of Rogaland, Norway. The HRQL was measured by the Nottingham health profile (NHP). At both evaluations clinical and demographic variables were determined during semistructured interviews and by clinical examinations by a neurologist. RESULTS: During the 4 year follow up period there was a significant increase in NHP scores, reflecting a decreased HRQL, in the dimensions of physical mobility, emotional reactions, pain, and social isolation. In the same time period mean total NHP score increased from 120.0 (SD 102.6) to 176.0 (SD 119.4) (p<0.01). There were no clinical or demographic factors found in 1993 that identified patients at higher risk for developing decreased HRQL. Increased UPDRS score (unified Parkinson's disease rating scale) and Hoehn and Yahr stage during the 4 year study period correlated with increased NHP scores. Even though there was no increase in depressive symptoms or self reported insomnia, these symptoms, together with lower Schwab and England score, were the most important factors for a poor HRQL in 1997. CONCLUSIONS: Parkinson's disease has a substantial impact on HRQL. Despite modern care, we found a significantly increased distress during the 4 year follow up period. Increased parkinsonism, measured by UPDRS and Hoehn and Yahr stage, correlated with increased stress, not only in the dimension of physical mobility, but also in the areas of pain, social isolation, and emotional reactions. In addition to the clinical examination, HRQL scoring provides valuable information on the total health burden of Parkinson's disease in both cross sectional and longitudinal evaluations, and contributes to a more comprehensive picture of the total disease impact. PMID- 11032609 TI - Systematic review of acute levodopa and apomorphine challenge tests in the diagnosis of idiopathic Parkinson's disease. AB - OBJECTIVES: To perform a systematic review of studies examining the diagnostic accuracy of acute challenge tests with levodopa and/or apomorphine in parkinsonian syndromes to assess their value in the diagnosis of idiopathic Parkinson's disease. METHODS: A literature search including Medline and the Cochrane Library was performed for studies published in any language comparing acute levodopa and/or apomorphine response with chronic levodopa therapy in parkinsonian syndromes. Abstracted sensitivity and specificity data were summarised using variance weighting and conditional logistic regression for studies comparing two challenge tests. RESULTS: Thirteen studies were located: four examining de novo patients and nine examining patients with well established idiopathic Parkinson's disease and non-parkinsonian conditions. Despite the significant heterogeneity in the methodologies employed, the comparable results suggest that this had little effect on the accuracy of the tests. The sensitivity for the diagnosis of established idiopathic Parkinson's disease was: apomorphine 0.86 (95% confidence interval (95% CI) 0.78-0.94), acute levodopa 0.75 (95% CI 0.64-0.85), and chronic levodopa therapy 0.91 (95% CI 0.85-0.99). The specificity for the diagnosis of established idiopathic Parkinson's disease was: apomorphine 0.85 (95% CI 0.74-0.96), acute levodopa 0.87 (95% CI 0. 77-0.97), and chronic levodopa therapy 0.77 (95% CI 0.61-0.93). The number of patients positive for each test divided by the number with clinically diagnosed de novo disease was: apomorphine 0.63 (95% CI 0. 56-0.70), acute levodopa 0.69 (95% CI 0.59-0.80), and chronic levodopa therapy 0.76 (95% CI 0.70-0.82). CONCLUSIONS: The accuracy of the acute levodopa and apomorphine challenge tests is similar to, but not superior than, that of chronic levodopa therapy in the diagnosis of idiopathic Parkinson's disease. As most patients will be given chronic dopamimetic therapy, these tests add nothing while causing significant adverse events and additional cost. PMID- 11032610 TI - Prognosis of stroke in the south of Greece: 1 year mortality, functional outcome and its determinants: the Arcadia Stroke Registry. AB - OBJECTIVES: For Greece, data on incidence of stroke, type of stroke, and prognosis of stroke is limited. Recently, results on incidence of stroke were published. Here 1 year mortality, functional outcome after a first ever stroke, and determinants of the prognosis are described. METHODS: A population based registry was established in the Arcadia area, located in eastern central Peloponessos in southern Greece. Between 1 November 1993 and 31 October 1995, 555 patients with a first ever stroke were identified using information from death certificates, hospital records, public health centres and general practitioners. Extensive information on cardiovascular risk factors and stroke characteristics was obtained. After 1 year a modified Rankin score was determined in all surviving patients. RESULTS: After 1 year of follow up, 204 (36.8%) patients died. The probability of survival 1 year after stroke was higher for cerebral infarction than for intracerebral haemorrhage; 67.8% (95% confidence interval (95% CI) 64-72) and 46.4% (35-57), respectively. Of the survivors, 68.9% had either no symptoms or symptoms that would not interfere with their capacity to look after themselves (Rankin score 0 to 2). Increasing age and low Glasgow coma scale score were the most powerful predictors of death within 1 year (p<0.01), whereas increasing age, atrial fibrillation, and low Glasgow coma scale score were the most important predictors of functional outcome 1 year after a stroke (p<0.01). CONCLUSIONS: One year mortality from stroke in Greece is similar to that of other industrialised countries. The most important factors that affect the prognosis of a patient with a first ever stroke are increasing age, stroke severity, and atrial fibrillation. PMID- 11032611 TI - Incidence of intracerebral and subarachnoid haemorrhage in southern Sweden. AB - OBJECTIVES: Spontaneous intracranial haemorrhage-that is, mainly subarachnoid haemorrhage (SAH) and primary intracerebral haemorrhage (PICH)-constitutes an important part of all strokes. As previous epidemiological studies have demonstrated highly variable incidence rates, we conducted a large prospective investigation of all haemorrhagic strokes during a 1 year period. METHODS: Twelve hospitals serving a defined population of 1.14 million in southern Sweden registered all cases with spontaneous intracranial haemorrhage, including those found dead outside hospitals, during 1996. All patients were examined with CT of the brain or underwent necropsy. Incidence rates adjusted to the Swedish population for age and sex, as well as location of haematoma and prevalence of risk factors were calculated. RESULTS: A total of 106 patients with SAH and 341 patients with PICH were identified. The annual incidence/100 000 was 10.0 (6.4 for men and 13.5 for women) for SAH and 28.4 (32.2 for men and 24.7 for women) for PICH when adjusted to the Swedish population. Subarachnoid haemorrhage affected twice as many women as men. The incidence of both types of haemorrhage increased with advancing age, but in particular, this was the case for supratentorial PICH. Lobar haematomas were the most common (51.6%) type of PICH. Among patients with PICH, 37% had hypertension, 41% other vascular disease, and 12% were on oral anticoagulation. Among patients with SAH, 28% had hypertension and 18% vascular disease before the haemorrhage but no one was on treatment with oral anticoagulation. CONCLUSIONS: The incidence of PICH was high, especially for the older age groups. PICH was, on average, three times as common as SAH. The study underscores the importance of PICH and SAH as significant stroke subgroups. PMID- 11032612 TI - Cognitive impairments after surgical repair of ruptured and unruptured aneurysms. AB - OBJECTIVES: To determine the frequency and severity of neuropsychological impairments associated with aneurysmal subarachnoid haemorrhage, and associated with repair of intracerebral aneurysms. METHODS: Two groups of patients who underwent repair of intracerebral aneurysms were studied: patients with unruptured aneurysms (n=20) and patients with ruptured aneurysms (n=27). All patients were administered a battery of standardised neuropsychological tests about 3 months after surgery. A subset of 12 patients with unruptured aneurysms were administered the battery both before and after elective repair of the aneurysm(s). A subset of six patients with ruptured aneurysms were given the test at both 3 months and 1 year after surgery. RESULTS: As previously reported for patients with ruptured aneurysms, patients with both ruptured and unruptured aneurysms performed, as a group, significantly below published norms on many of the neuropsychological tests after surgery. However, there were significant differences between preoperative and postoperative performance in the unruptured aneurysm group only on a few tests: measures of word fluency, verbal recall, and frontal lobe function. Performance of patients with ruptured aneurysms was significantly below that of patients with unruptured aneurysms only on a few tests of verbal and visual memory. In addition, group differences compared with published norms reflected severely impaired performance by a minority of patients, rather than moderately impaired performance in a majority of patients. CONCLUSIONS: Although patients who undergo repair of ruptured aneurysms perform, as a group, below published norms on many neuropsychological tests, significant impairments are seen in a minority of patients. Some of the impairments are associated with subarachnoid haemorrhage, whereas others (found in patients who underwent repair of unruptured aneurysms) are due to general effects of neurosurgery and perioperative management. Finally, some of the postoperative deficits are merely a reflection of premorbid weaknesses. PMID- 11032613 TI - Ultrasonic evaluation of pathological brain perfusion in acute stroke using second harmonic imaging. AB - OBJECTIVE: To evaluate the use of transient response second harmonic imaging (HI) by means of ultrasound to assess abnormalities of cerebral echo contrast agent enhancement in patients with acute stroke. METHODS: The study comprised 25 patients with acute onset of hemispheric stroke (<24 h) with sufficient insonation conditions and 14 control subjects without cerebrovascular disease. All stroke patients had HI, extracranial and transcranial colour coded duplex examinations of the arteries supplying the brain, and clinical examinations (European stroke scale) performed in the acute phase, on day 2, and within 1 week. Acute CT was repeated within 1 week and facultatively accompanied by angiography. Examinations using HI were performed in an axial diencephalic plane of section using the transtemporal acoustic bone window. After bolus application of galactose based microbubbles, 61 ultrasound images with a cardiac cycling triggering frequency of once every 2 seconds were recorded and evaluated off line. Focal perfusion deficit was identified if no contrast enhancement was visualised in a circumscribed region of interest and insufficient temporal bone window was excluded. In cases of reappearance of contrast enhancement reperfusion was assessed. RESULTS: Adequate cerebral contrast enhancement could be seen in 21 subjects. In seven, a large hemispheric deficit of contrast enhancement affecting the entire middle cerebral artery (MCA) territory was detectable; the lentiform nucleus was affected in three subjects. Assessment of cerebral contrast abnormalities was possible in two patients with superficial MCA infarctions but in none of the patients with lacunar ischaemias. None of the control persons had focal deficits of cerebral echo contrast enhancement. In all patients with complete MCA infarction and striatocapsular infarction, presumed ischaemic areas in HI examinations correlated with final CT findings. Overall sensitivity and specifity of HI examinations for predicting size and localisation of the infarction were 75 and 100%, respectively. During follow up, reappearance of contrast enhancement was determined in three patients, in two patients circulatory arrest due to malignant brain oedema with missing contrast enhancement in the entire cerebral hemisphere could be seen. Extent of contrast enhancement deficits significantly correlated with the clinical status on admission and after 1 week (p<0.01). CONCLUSIONS: Second harmonic imaging is the first ultrasonic technique that enables visualisation of pathological cerebral echo contrast enhancement. Because this method identifies deficits of focal contrast enhancement in patients with acute stroke and allows estimation of the final infarct size and clinical prognosis, it may help to select and monitor patients for invasive therapies. PMID- 11032614 TI - Comparison of the pattern of atrophy of the corpus callosum in frontotemporal dementia, progressive supranuclear palsy, and Alzheimer's disease. AB - OBJECTIVES: The loss of the neurons in layer 3, one of the groups of cortical neurons most vulnerable in various degenerative brain diseases, results in axonal degeneration leading to atrophy of the corpus callosum. Previous studies showed callosal atrophy in three degenerative dementias: frontotemporal dementia (FTD), progressive supranuclear palsy (PSP), and Alzheimer's disease (AD). However, it is unclear whether a characteristic pattern of atrophy is present in each. The objective of this study was to investigate whether the pattern of the callosal atrophy was different among patients with FTD, PSP, or early onset AD. METHODS: Eleven patients with FTD, nine patients with PSP, 16 patients with early onset AD, and 23 normal controls, all age and sex matched, were studied using MRI. The ratios of midsagittal corpus callosum areas to the midline internal skull surface area on T1 weighted images were analyzed. The corpus callosum was divided into quarters: the anterior, middle-anterior, middle-posterior, and posterior portions. RESULTS: Compared with controls, all three patient groups had significantly decreased total callosal/skull area ratio. An analysis of covariance adjusted for the total callosal area/skull area ratio showed that the anterior quarter callosal/skull area ratio in FTD, the middle-anterior quarter area ratio in PSP, and the posterior quarter area ratio in AD were significantly smaller than those in the other three groups. CONCLUSION: Although atrophy of the corpus callosum is not specific to any degenerative dementia, the patterns of the atrophy are different among patients with FTD, PSP, or early onset AD. Differential patterns of callosal atrophy might reflect characteristic patterns of neocortical involvement in each degenerative dementia. PMID- 11032615 TI - Visual rating and volumetry of the medial temporal lobe on magnetic resonance imaging in dementia: a comparative study. AB - OBJECTIVES: It has been shown that atrophy of medial temporal lobe structures such as the hippocampus and entorhinal cortex shown on MRI may distinguish patients with Alzheimer's disease from healthy controls. However, the diagnostic value of visual inspection and volumetry of medial temporal lobe atrophy (MTA) on MRI in a clinical setting is insufficiently known. METHODS: Medial temporal lobe atrophy in 143 patients was visually rated from hard copies, using a 0-4 rating scale and a comparison was made with the volumes (cm(3)) of the medial temporal lobe as estimated with volumetry, using a stereological method. All patients were recruited in an unselected way in a clinical setting in the centre for memory impairments at the Huddinge University Hospital. Patients with Alzheimer's disease (n=41), patients with other dementias (vascular dementia, frontotemporal dementia, and unspecified dementia; n=36) as well as non-demented subjects (n=66) were included. Medial temporal atrophy and volumetry were evaluated as a diagnostic tool by performing logistic regression analysis including age, sex, and mini mental state examination (MMSE) score and calculating the sensitivity and specificity and percentage correct classification. RESULTS: Visual and volumetric analysis yielded statistically significant differences between patients with Alzheimer's disease and non-demented subjects, as well as between those with other dementias and non-demented subjects. Combining MMSE scores and visually rated MTA ratings yielded a sensitivity of 95% for Alzheimer's disease, 85% for other dementias. Non-demented subjects were identified with a specificity of 96%. Volumetry did not have an added value over the MMSE score alone. CONCLUSIONS: Visual rating of MTA is a clinically useful method for differentiating Alzheimer's disease from controls and is both quicker and more accurate than volumetry. PMID- 11032616 TI - Effects of topiramate on cognitive function. AB - OBJECTIVE: To explore the impact of topiramate on tests of intellect and other cognitive processes. METHODS: This was a retrospective study. The neuropsychological test scores of 18 patients obtained before and after the introduction of treatment with topiramate (median dose 300 mg) were compared with changes in test performance of 18 patients who had undergone repeat neuropsychological assessments at the same time intervals. Complaints of cognitive decline precipitated referral for reassessment in five cases in the topiramate treated group. The groups were matched for age and intellectual level at the time of the first assessment. Patients were assessed using the WAIS-R, tests of verbal and non-verbal memory, language, and perceptual processing. A subgroup of patients underwent a brief reassessment after the withdrawal or substantial reduction of topiramate. RESULTS: Repeat assessments in those taking topiramate were associated with a significant deterioration in many domains, which were not seen in the comparison group. The greatest changes were for verbal IQ, verbal fluency, and verbal learning (p<0. 001). Improvements in verbal fluency (p<0.05), verbal learning (p<0. 01), and digit span (p<0.001) were recorded in those patients who had topiramate withdrawn or reduced. CONCLUSIONS: In our patient group topiramate had a negative impact on cognition which was consistent with subjective complaints of patients. Tests requiring verbal processing seemed especially sensitive to the drug. A decline in verbal intellect (VIQ), a measure which has been considered by some to be insensitive to antiepileptic drug effects, was particularly striking. Caution is warranted in the interpretation of the findings due to methodological limitations of the study design. Further investigation of mediating factors such as serum concentrations, comedication, and other potential risk factors, however, is needed to enable appropriate targeting of treatment with this effective antiepileptic agent. PMID- 11032617 TI - William Stewart Halsted (1852-1922). PMID- 11032618 TI - Idiopathic ocular neuromyotonia: a neurovascular compression syndrome? AB - Ocular neuromyotonia in the muscles innervated by the right oculomotor nerve was diagnosed in a patient without a history of radiation therapy. Electromyography of the levator palpebrae showed continuous motor unit activity. Brain MRI disclosed a close contact between the right third cranial nerve and a basilar artery dolichoectasia. The patient partly benefited from carbamazepine therapy. This unique finding suggests that neurovascular compression syndrome could be an hitherto unrecognised cause of ocular neuromyotonia. PMID- 11032619 TI - Misidentification syndromes related to face specific area in the fusiform gyrus. AB - The "delusional misidentification syndromes" are a group of uncommon and varied disorders in which, in typical form, the patient thinks that a particular familiar person is someone else or a certain familiar place is a duplicate. Although first identified and considered a memory disorder by Pick, evidence in support of this has been difficult to identify. They have been most often seen in various psychotic and organic brain diseases but lesions have been generally diffuse although the right temporal lobe has been implicated. A patient was investigated who abruptly developed a disorder wherein she misidentified her husband as her deceased sister and claimed that her home was a duplicate of her real home that were typical of Fregoli syndrome and Pick's reduplicative paramnesia, respectively. A discrete area of brain damage, probably ischaemic, in this patient was seen on MRI in the anterior part of the right fusiform gyrus and a smaller area in the nearby anterior middle and inferior temporal gyri with associated parahippocampal and hippocampal atrophy. A high order nervous system function that is devoted to the identification of faces is located in the adjacent midportion of the fusiform gyrus and a similar locus for environmental scenes, termed the parahippocampal place area, is present in the bordering parahippocampal gyrus. The misidentification phenomena in this case can be explained by disruption of the connections of these highly specialised areas with the most anterior inferior and medial part of the right temporal lobe where long term memory and mechanisms for the retrieval of information that are required for the visual recognition of faces and scenes are stored. PMID- 11032620 TI - Characteristics of falling in patients with stroke. AB - OBJECTIVE: To identify the risk factors for falling after stroke, to establish the relation with lesion localisation, and to evaluate the incidence of falling. METHODS: The falling history and the mood of 293 patients with stroke were investigated by a standard questionnaire. Other information (time since stroke, risk factors, and CT) about patients was obtained from their hospital records. RESULTS: Increasing age, depression, and heart disease were significant risk factors for falling (heart disease had a negative influence). A right hemispheric infarct was significantly more common among the falling group. CONCLUSION: This study suggests clues for possible modifications of the management of patients with stroke during the recovery period. PMID- 11032621 TI - Description of a simple test for CADASIL disease and determination of mutation frequencies in sporadic ischaemic stroke and dementia patients. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare inherited adult onset disease characterised most commonly by cerebral ischaemic events and dementia. It is caused by mutations in the Notch3 gene with most clustering in exons 3 and 4. Whether these mutations have any influence on common sporadic ischaemic stroke or dementia cases has not been investigated, partly hampered by the lack of a readily usable genetic test. An easy to use diagnostic array for CADASIL was designed using various restriction endonucleases for the known mutations in exons 3 and 4 and novel mismatch primers were designed where no such enzymes existed. This array was used to identify the allele frequencies of CADASIL mutations and polymorphisms in selected disease cohorts. Seventy patients with radiologically established sporadic ischaemic stroke and 77 patients from a specialist young dementia clinic were recruited. One hundred and seventeen age and sex matched asymptomatic controls were also identified. The diagnostic array was found to work well. None of the 14 known mutations and three previously identified polymorphisms (C474A, A587G, and C594A) in exons 3 and 4 were present in 140 stroke, 110 dementia, or 234 control chromosomes. Molecular variant C381T occurred with a higher frequency of 0.13, whereas G684A occurred with a lower frequency (0.09) than previously reported, although there were no statistical differences between selected cohorts. In conclusion, a readily usable genetic test for CADASIL has been devised that was used to determine allele frequencies in well characterised cohorts of sporadic stroke and dementia patients. The data suggest that despite the clinical resemblance, CADASIL is not a common masquerading cause of stroke or dementia. The test will enable units locally to rapidly screen patients with suspected CADASIL. PMID- 11032622 TI - Unusual muscle pathology in McLeod syndrome. AB - Muscle pathology in McLeod syndrome is usually mild; patchy necrotic or regenerating fibres, occasional internal nuclei, and the absence of an inflammatory cell infiltrate are the usual findings. We report on a 29 year old man presenting with chronic fatiguability and excessive sweating in whom an open quadriceps muscle biopsy demonstrated grouped necrotic fibres accompanied by striking patchy mononuclear cell infiltrates. The diagnosis of McLeod syndrome was made on the basis of red blood cell acanthocytosis, raised serum creatine kinase, and weak expression of Kell blood group antigens. The quadriceps muscle infiltrate consisted principally of histologically typical macrophages. These cells had prominent nucleoli, displayed numerous mitoses, and were strongly CD68+. A small population of typical CD3+, CD43+ lymphocytes was also present. In addition, a small population of large atypical CD3+ cells was noted. Immunoperoxidase stains for CD20, CD30, CD79a, and CD56 were negative. Immunocytochemical studies for the common muscular dystrophies were normal. The muscle biopsy findings highlight a potential for confusion of this condition with idiopathic polymyositis. The expanding range of muscle pathology reported in McLeod syndrome, to which this case adds, may reflect variable involvement of the XK gene on chromosome Xp21, or of the adjacent loci of Duchenne muscular dystrophy and chronic granulomatous disease. PMID- 11032623 TI - Effect of prolonged neck muscle vibration on lateral head tilt in severe spasmodic torticollis. AB - Short term vibration of the dorsal neck muscles (10-35 s) is known to induce involuntary movements of the head in patients with spasmodic torticollis. To investigate whether neck muscle vibration might serve as a therapeutic tool when applied for a longer time interval, we compared a vibration interval of 5 seconds with a 15 minute interval in a patient with spasmodic torticollis with an extreme head tilt to the right shoulder. Head position was recorded with a two camera optoelectronic motion analyzer in six different test conditions. Vibration regularly induced a rapid change of head position that was markedly closer to a normal, upright posture. After 5 seconds of vibration, head position very quickly returned to the initial position within seconds. During the 15 minute interval, head position remained elevated. After terminating vibration in this condition, the corrected head position remained stable at first and then decreased slowly within minutes to the initial tilted position. CONCLUSIONS: (1) In this patient, muscle vibration was the specific sensory input that induced lengthening of the dystonic neck muscles. Neither haptic stimulation nor transcutaneous electrical stimulation had more than a marginal effect. (2) The marked difference in the change of head position after short and prolonged stimulation supports the hypothesis that spasmodic torticollis might result from a disturbance of the central processing of the afferent input conveying head position information-at least in those patients who are sensitive to sensory stimulation in the neck region. (3) Long term neck muscle vibration may provide a convenient non-invasive method for treating spasmodic torticollis at the central level by influencing the neural control of head on trunk position. PMID- 11032624 TI - Alzheimer's disease and frontotemporal dementia are differentiated by discriminant analysis applied to (99m)Tc HmPAO SPECT data. AB - OBJECTIVE: Alzheimer's disease (AD) and frontotemporal dementia (FTD) are the most frequent neurodegenerative cognitive disorders. However, FTD remains poorly recognised clinically. The use of (99m)HmPAO-single photon emission computed tomography (SPECT) has been demonstrated in the differentiation of AD and FTD. Nethertheless, there are very few comparative studies designed to assess its precise value in this differential diagnosis. The aim was to determine a simple decision rule, deduced from statistical analysis, which, if applied to regions of interest (ROIs) and mini mental state examination (MMSE), could improve the predictive value of SPECT in differential diagnosis between AD and FTD. METHODS: Forty patients, 20 with probable AD and 20 with probable FTD were included. All patients underwent brain SPECT imaging, after an intravenous injection of (99m)Tc HmPAO-(555mBq). For each patient, 20 ROIs were determined on the Fleishig's slice and their activity was normalised to the mean cerebellar activity. Bivariate analysis (Wilcoxon rank tests) and multivariate analysis (stepwise discriminant analysis) were performed to determine the subgroup of variables able to give the highest predictive value for this differential diagnosis. A simple decision rule was built from a predictive score derived by factorial discriminant analysis. RESULTS: As previously described, the fixation defect was found in frontal regions of interest (ROIs) in FTD and in the left temporoparietal-occipital ROIs in AD. Among the 21 variables, five were finally selected: right median frontal, left lateral frontal, left tempoparietal, left temporoparietal-occipital areas, and MMSE. One hundred per cent of patients with FTD were correctly classified by the decision rule (20/20 patients) and 90% of patients with AD (18/20). CONCLUSION: AD and FTD are differentiated by SPECT. Automatic classification based on a decision rule deduced from factorial discriminant analysis could enhance its performance. PMID- 11032626 TI - Does APO epsilon4 correlate with MRI changes in Alzheimer's disease? AB - OBJECTIVE: To assess the relation between APO E genotype and MRI white matter changes in Alzheimer's disease. The APO epsilon4 allele is correlated with amyloid angiopathy and other neuropathologies in Alzheimer's disease and could be associated with white matter changes. If so, there should be a dose effect. METHODS: 104 patients with probable Alzheimer's disease (NINCDS-ADRDA criteria) in this Alzheimer's Disease Research Centre were studied. Patients received MRI and APO E genotyping by standardised protocols. Axial MRI was scored (modified Schelten's scale) for the presence and degree of white matter changes and atrophy in several regions by a neuroradiologist blinded to genotype. Total white matter and total atrophy scores were also generated. Data analysis included Pearson's correlation for regional and total imaging scores and analysis of variance (ANOVA) (or Kruskal-Wallis) and chi(2) for demographic and disease related variables. RESULTS: 30 patients had no epsilon4, 53 patients were heterozygous, and 21 patients were homozygous. The three groups did not differ in sex distribution, age of onset, age at MRI, MMSE, clinical dementia rating, or modified Hachinski ischaemia scores. There were no significant correlations between total or regional white matter scores and APO E genotype (Pearson correlation). CONCLUSIONS: No correlation between total or regional white matter scores and APO E genotype was found. The pathogenesis of white matter changes in Alzheimer's disease may be independent of APO E genotype. PMID- 11032627 TI - Carotid brainstem reflex myoclonus after hypoxic brain damage. AB - A patient comatose after acute anoxia developed bilaterally synchronous, periodic myoclonic jerks most prominently in the bilateral upper limbs. Although the myoclonus seemed to occur spontaneously, electrophysiological studies showed that the myoclonic jerks correlated in timing and size with arterial pulses, and was suppressed by massage over the carotid sinus. It is proposed that the present myoclonus is a variant of brainstem reflex myoclonus in which arterial pulses served as intrinsic trigger stimuli via the carotid sinus and the medullary reticular formation. PMID- 11032625 TI - Altered glycosylation of acetylcholinesterase in lumbar cerebrospinal fluid of patients with Alzheimer's disease. AB - As clinical diagnosis of Alzheimer's disease is only 80%-90% accurate, there is a need to identify biochemical markers of Alzheimer's disease. Previous studies have shown an abnormality in the glycosylation of acetylcholinesterase (AChE) in the CSF collected postmortem from patients with Alzheimer's disease. This abnormality was very specific for Alzheimer's disease, as it was not detected in other illnesses causing dementia. We report here that the glycosylation of AChE is also altered in lumbar CSF collected antemortem. The altered glycosylation was due to increased concentrations of a minor AChE isoform that does not bind to concanavalin A (Con A). Glycosylation of AChE may eventually be of diagnostic value, especially when used in combination with other CSF markers. PMID- 11032628 TI - Atypical form of non-Langerhans histiocytosis with disseminated brain and leptomeningeal lesions. AB - An 18 year old girl presented with acute visual loss. T2 weighted brain MRI showed areas of hyperintensities in the thalamic nuclei, internal capsule, lentiform nuclei, the subarachnoidal spaces, and a retrobulbar infiltration. Analysis of CSF showed numerous foamy histiocytes without malignant cells, raised protein, and depressed glucose concentration. Biopsy of the right thalamus demonstrated aggregates of histiocytes with immunohistological and ultrastructural characteristics of non-Langerhans cell histiocytosis. The patient improved with chemotherapy and corticosteroids. After 3 months of treatment, CSF analysis showed no more histiocytes. Cytological examination of CSF can be helpful for the management of patients with extensive histiocytic infiltration. PMID- 11032629 TI - Contralateral hearing loss as an effect of venous congestion at the ipsilateral inferior colliculus after microvascular decompression: report of a case. AB - Contralateral hearing loss after surgical procedures within the cerebellopontine angle is rarely seen and its pathophysiological background is not yet understood. A patient with contralateral hearing loss after microvascular decompression for trigeminal neuralgia is described. Ipsilateral brainstem auditory potential (BAEP) monitoring and facial nerve EMG did not show major abnormalities. During otherwise uneventful and successful surgery a branch of the petrosal vein was sacrificed to widen the access to the trigeminal root exit zone. On the third postoperative day the patient complained about contralateral hearing loss, which was verified by audiometry. Contralateral BAEPs showed low amplitudes and delayed interpeak latencies. Brain CT was normal. Brain MRI on the 8th postoperative day disclosed abnormal signals within the ipsilateral inferior colliculus. Intravenous heparinisation was performed and hearing slowly recovered over a 3 month period. Results from this patient offer a pathophysiological mechanism for contralateral hearing loss after cerebellopontine angle surgery, illustrate the importance of venous drainage preservation, gives evidence about the generation of BAEP components within the contralateral brainstem, and stresses the importance of intraoperative BAEP monitoring. PMID- 11032630 TI - Bilateral stenting of symptomatic and asymptomatic internal carotid artery stenosis due to fibromuscular dysplasia. AB - Stent grafting of internal carotid artery (ICA) stenoses due to fibromuscular dysplasia has been rarely and only unilaterally carried so far. Bilateral carotid stent grafting of ICA stenoses due to fibromuscular dysplasia has not been reported previously. In a 37 year old woman with recurrent right hemispheric transitory ischaemic attacks, a non-disabling minor stroke, and recurrent right amaurosis fugax despite antithrombotic therapy, cerebral angiography disclosed a long segment narrowing, distal, high grade (95%) stenosis of the right ICA and a long narrowing, distal high grade (70%) stenosis of the left ICA. Morphological features of both stenoses were indicative of fibromuscular dysplasia. The right sided stenosis was stented with a PTFE-HEMOBAHN endoprosthesis; this was followed by a brief, postprocedural left sided hemiparesis. The left sided ICA stenosis was successfully stented by the same procedure. Nine months later, both stents were still patent and the patient was symptom free. Bilateral carotid stenting may remain an alternative to endarterectomy in bilateral ICA stenosis due to fibromuscular dysplasia when ischaemic events persist despite full antithrombotic therapy. PMID- 11032631 TI - Lead poisoning from complementary and alternative medicine in multiple sclerosis. AB - A patient with multiple sclerosis is described who was treated for neurological symptoms thought to be a progression of his disease but subsequently found to be caused by lead poisoning secondary to the use of alternative medicine. His clinical signs improved with oral chelation therapy. Neurologists should consider asking about the use of complementary and alternative medicine before simply attributing symptoms and signs to exacerbation of multiple sclerosis. PMID- 11032632 TI - Spinal cord schistosomiasis. PMID- 11032633 TI - The rostrocaudal gradient for somatosensory perception in the human postcentral gyrus. PMID- 11032634 TI - Crossed face apraxia. PMID- 11032635 TI - A case of Bickerstaff's brainstem encephalitis mimicking tetanus. PMID- 11032636 TI - Lysozyme in ventriculitis: a marker for diagnosis and disease progression. PMID- 11032637 TI - Clinicoanatomical correlates of a Fou rire prodromique in a pontine infarction. PMID- 11032638 TI - Proximal median mononeuropathy associated with an anomalous deep course through the brachialis muscle. PMID- 11032639 TI - Documented growth of a temporal arachnoid cyst. PMID- 11032640 TI - Pituitary apoplexy presenting as massive subarachnoid hemorrhage. PMID- 11032641 TI - Early detection of non-compliance in Wilson's disease by consecutive copper determination in cerebrospinal fluid. PMID- 11032642 TI - Systemic effects of intranasal steroids: an endocrinologist's perspective. AB - Intranasal steroids (INSs) are established as first-line treatment for allergic rhinitis. Extensive use of INSs with few reported adverse events supports the safety of these medications. Nevertheless, the prescription of more potent INSs for consistent and more prolonged use to younger and older patients, often in combination with inhaled corticosteroids, justifies the careful examination of their potential adverse systemic effects. Systemic bioavailability of INSs, by way of nasal and intestinal absorption, can be substantial; but current INSs vary significantly in their degree of first-pass hepatic inactivation and clearance from the body of the swallowed drug. For safety studies of INSs, distinguishing detectable physiologic perturbations from important adverse events is aided by an understanding of normal endocrine physiology and the methods used to test these systems. A review of available information indicates that (1) sensitive tests can measure the effects of INSs on biologic feedback systems, but they do not accurately predict clinically relevant adverse effects; (2) the primary factors that influence the relationship between therapeutic and adverse systemic effects of INSs are dosing frequency and efficiency of hepatic inactivation of swallowed drug; (3) INS treatment in recommended doses does not cause clinically significant hypothalamic-pituitary-adrenal axis suppression; (4) growth suppression can occur with twice-daily administration of certain INSs but does not appear to occur with once-daily dosing or with agents with more complete first-pass hepatic inactivation; (5) harmful effects of INSs on bone metabolism have not yet been adequately studied but would not be expected with the use of an INS dose and dosing frequency that do not suppress basal hypothalamic-pituitary adrenal axis function or growth; and (6) these conclusions apply to INS treatment alone and in recommended doses-the risk of adverse effects in individual patients who are treated with INSs is increased by excessive dosing or concomitant inhaled corticosteroid or other topical corticosteroid therapy. PMID- 11032643 TI - Increased signalling effort when survival prospects decrease: male-male competition ensures honesty. AB - For signalling to be honest the handicap principle claims that signals must impose fitness costs so that only the best individuals can afford the most exaggerated signals. The cost of signalling in terms of reduced survival decreases, however, towards the end of an individual's lifetime, which can induce an increase in signalling effort as a terminal effort. I show for the three spined stickleback, Gasterosteus aculeatus, that a decrease in survival prospects through impaired condition leads to an increase in red nuptial coloration that makes the signal less reliable as an indicator of male parental ability. Males in poor condition with a large signal sometimes cannibalized all the eggs they received, probably to start a new breeding cycle with a higher energy reserve. However, the inclusion of socially imposed costs of signalling through male-male competition during courtship increased the honesty of the signal, as some males in poor condition and of poor parental ability decreased their signal expression. Some cheaters still occurred, but the signalling system was honest on average. This implies that socially imposed costs are important in the maintenance of honest sexual signalling. Dishonesty may occur under favourable conditions when the cost of signalling is reduced. This emphasizes the importance of considering the environmental conditions experienced by individuals when investigating the evolution and maintenance of honest sexual signals. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032644 TI - Reproductive stage and history affect the phonotactic preferences of female midwife toads, Alytes muletensis. AB - We investigated whether the reproductive stage or reproductive history of females affects their preferences for male calls that differ in frequency. We repeatedly tested gravid female midwife toads prior to and immediately after mating, and during ovulation. When females were ovulating they were much more consistent in their choice of call than when they were gravid but not ovulating. When females had recently mated they still showed positive phonotaxis, but did not reliably discriminate between alternative calls. This study is the first to test gravid female anurans repeatedly at different stages in their reproductive cycle (ovulating or not; pre- and postmating). It highlights the need to treat with caution the results from population-based preference studies in which individuals are tested only once, and in which variation in reproductive state is not taken into account. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032645 TI - Male provisioning is negatively correlated with attempted extrapair copulation frequency in the stitchbird (or hihi). AB - In species with biparental care of offspring and high levels of extrapair parentage, paired males may suffer reduced fitness by investing in offspring that are not their own. In such instances, males are predicted to evolve some form of discrimination between kin and nonkin and vary investment accordingly. One simple form of discrimination is to follow behavioural cues to paternity. We investigated paternal contribution to chick rearing in a socially monogamous population of stitchbirds, Notiomystis cincta, with high levels of extrapair copulation (EPC) and extrapair paternity. The relative contribution of the male to chick provisioning was negatively correlated with attempted EPC frequency. Because the majority of EPC attempts are forced and conspicuous in stitchbirds they are an obvious cue for males to use. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032646 TI - Food choice behaviour may promote habitat specificity in mixed populations of clonal and sexual Potamopyrgus antipodarum. AB - Genetic polymorphism along an environmental gradient may be maintained if disruptive selection on habitat-specific traits leads to a correlated response in traits that reduce gene flow between habitats. We studied a short-distance cline in a population of freshwater snails Potamopyrgus antipodarum in which sexual and clonal snails coexist. Sexuals and clones show a life history cline by depth: snails reproduce at a smaller size in shallower habitats. Clones are also structured genetically across habitats and seem not to mix, even though habitats are within the dispersal distance of the snails and the opportunity for gene flow via migration must be considerable. Because habitat preference may promote divergence in both clones and sexuals along the depth gradient, we investigated whether snails show habitat-specific food choice behaviour that could reduce migration. We tested the food choice behaviour of the snails by exposing them simultaneously to food from their home and adjacent habitats. Both juvenile and adult snails from the shallow shore bank and a mid-water macrophyte habitat preferentially grazed on the vegetation of their original habitats. We suggest that the observed genetic and life history cline may be maintained by food choice behaviour that may promote a partial barrier to gene flow between the habitats. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032647 TI - Delayed benefits of paternal care in the burying beetle Nicrophorus vespilloides. AB - Burying beetles, Nicrophorus spp., inter the carcasses of small vertebrates as a food source for their offspring. Females can bury a carcass and rear a brood on it alone, but are frequently assisted by a male whose presence reduces the risk of the carcass being taken over by other beetles. However, the male often stays for longer than the carcass is vulnerable to take-over, and he cares for the brood without conferring any further benefits on it. In a laboratory experiment using N. vespilloides, we found that, in the absence of competitors, male assistance conferred no advantages on the brood for which he was caring, but significantly increased the subsequent reproductive success of his mate, in terms of the mass and rate of development of a second brood, reared alone. We suggest that this is due to a reduced parental effort of assisted females, who spent less time feeding offspring and more time resting than unassisted females whilst rearing their first broods. In the field, a female is unlikely to pair with the same male for consecutive broods, so we discuss the possible benefits a male may accrue from increasing his mate's reproductive success. We also discuss the relevance of these results to our understanding of the evolution of biparental care in birds. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032648 TI - Magnetic information calibrates celestial cues during migration. AB - Migratory birds use celestial and geomagnetic directional information to orient on their way between breeding and wintering areas. Cue-conflict experiments involving these two orientation cue systems have shown that directional information can be transferred from one system to the other by calibration. We designed experiments with four species of North American songbirds to: (1) examine whether these species calibrate orientation information from one system to the other; and (2) determine whether there are species-specific differences in calibration. Migratory orientation was recorded with two different techniques, cage tests and free-flight release tests, during autumn migration. Cage tests at dusk in the local geomagnetic field revealed species-specific differences: red eyed vireo, Vireo olivaceus, and northern waterthrush, Seiurus noveboracensis, selected seasonally appropriate southerly directions whereas indigo bunting, Passerina cyanea, and grey catbird, Dumetella carolinensis, oriented towards the sunset direction. When tested in deflected magnetic fields, vireos and waterthrushes responded by shifting their orientation according to the deflection of the magnetic field, but buntings and catbirds failed to show any response to the treatment. In release tests, all four species showed that they had recalibrated their star compass on the basis of the magnetic field they had just experienced in the cage tests. Since release tests were done in the local geomagnetic field it seems clear that once the migratory direction is determined, most likely during the twilight period, the birds use their recalibrated star compass for orientation at departure. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032649 TI - Coding in the song of the wren: importance of rhythmicity, syntax and element structure. AB - Communication between territorial songbirds usually involves a transfer of encoded information over long distances. We would expect coding and decoding strategies to be adaptive given the constraints imposed by the habitat. We used playback to examine some song parameters important for information transfer in the wren, Troglodytes troglodytes. Six stimuli were tested with various modifications to song rhythmicity, song composition, element structure, syntax and overall song spectra. Song features encoding information essential for eliciting a territorial response seemed to be embedded in the fine structure of song elements, that is, their variations in frequency and amplitude over time. To function, this strategy must be flexible enough to accommodate the deleterious effects of habitat-induced degradation. All stimuli composed of original song elements elicited responses regardless of their other alterations. Rhythmicity seemed not to be essential for eliciting territorial behaviour. However, alterations in song rhythmicity, syntax and spectra affected territorial responses, suggesting that these nonessential song parameters do contribute to the options for discrimination. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032650 TI - Mate fidelity and breeding site tenacity in a monogamous sandpiper, the black turnstone. AB - We examined the relationship between mate fidelity and breeding site tenacity during a 5-year study of the black turnstone, Arenaria melanocephala, a socially monogamous sandpiper breeding in subArctic Alaska. We tested the predictions of several hypotheses regarding the incidence of divorce and the benefits of fidelity to mate and breeding site. Interannual return rates to the breeding grounds (88% for males, 79% for females) were among the highest yet recorded for any scolopacid sandpiper, and 88% of returning birds nested on their previous year's territory. The annual divorce rate was only 11%, and mate fidelity was significantly linked to fidelity to territory but independent of sex and year. Males arrived in spring significantly earlier than their mates and interannual fidelity was influenced by the relative timing of arrival of pair members. Reunited pairs had significantly higher fledging success than new pairs formed after death or divorce. The incidence of divorce was unrelated to reproductive success the previous year, although birds nested significantly further away after failure than after a successful nesting attempt. Sightings of marked individuals suggested that members of pairs do not winter together, and breeding site tenacity provides a mechanism through which pair members can reunite. We reject the 'incompatibility' hypothesis for divorce in turnstones, and our data contradict predictions of the 'better option' hypothesis. Alternatively, we propose the 'bet-hedging' hypothesis to explain the occurrence of divorce, which transpires when an individual pairs with a new mate to avoid the cost of waiting for a previous mate to return. Such costs can include remaining unmated, if the former mate has died, or experiencing lower reproductive success because of delayed breeding. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032651 TI - The effect of social dominance on fattening and food-caching behaviour in Carolina chickadees, Poecile carolinensis. AB - Subordinates often have to wait for dominants to obtain food. As a result, their foraging success should be less predictable and they should therefore maintain a higher level of energy reserves compared with dominants. A corollary of this prediction is that subordinates should gain mass earlier in the day and maintain higher mass than dominants. We tested these predictions with captive Carolina chickadees. In two different experiments (one where birds were given ad libitum access to food and the other with food access limited to 60 min/day), we formed social flocks of two previously unfamiliar birds and compared their energy management (body fat and food caches) while they were in the flock with energy management when housed alone. Results from both experiments failed to support the predictions. Of all the parameters of body mass and food caching we measured only the following results were significant: (1) On the ad libitum food schedule, both subordinates and dominants accumulated more mass over the day when in a flock compared with when they were solitary, and there were no differences in mass gain between dominants and subordinates. (2) When analysed separately, dominants showed a higher evening mass in the flock compared with the solitary condition, a trend that runs opposite to the prediction. Our results suggest that when in favourable foraging conditions, social interactions might cause dominant and subordinate birds to accumulate more energy reserves as a result of competition. On the other hand, if food supply is limited, both dominants and subordinates may be forced to maintain similar fat reserves as an insurance against increased risk of starvation. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032652 TI - Behavioural isolation between two closely related Hawaiian Drosophila species: the role of courtship. AB - Behavioural isolation plays a critical role in several recent models of speciation. A detailed understanding of the process of speciation requires analysis of taxonomic groups that have not completed reproductive isolation. We studied D. silvestris and D. heteroneura because they are still in the process of divergence: behavioural isolation between them is incomplete, and neither postzygotic nor ecological isolation has been detected. Behavioural isolation is due to the failure of courtships between male D. silvestris and female D. heteroneura: there is no postzygotic isolation from either parental species. The F1 hybrids are as successful in courtship with parental individuals as same species pairs, which suggests that the hybrids resemble male D. heteroneura or female D. silvestris in some behaviour patterns that are crucial to mating success. We searched for this crucial resemblance by examining courtship between F1 hybrids and the parental adults. We found that successful F1 males are somewhat more similar than unsuccessful F1 males to D. heteroneura males, but nevertheless they were intermediate between males of the two species. We also found that in both species the presence of female wings is necessary for courtship to proceed to copulation. These results reinforce an earlier report that behavioural isolation between these species is largely attributable to the decision as to whether to court at all, rather than to the details of courtship. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032653 TI - Wasp predation and wasp-induced hatching of red-eyed treefrog eggs. AB - Eggs often suffer high levels of predation and, compared with older animals, embryos have few options available for antipredator defence. None the less, hatchlings can escape from many predators to which eggs are vulnerable. I studied early hatching as an antipredator defence of red-eyed treefrog embryos, Agalychnis callidryas, in response to egg predation by social wasps (Polybia rejecta). Red-eyed treefrogs attach their eggs to vegetation overhanging water, where they are exposed to arboreal and aerial predators. Wasps attacked half the egg clutches and killed almost a quarter of the eggs I monitored at a natural breeding site in Panama. Hatching tadpoles fall into the water, where they face aquatic predators. As predicted from improved survival of older hatchlings with aquatic predators, most undisturbed eggs hatched relatively late. However, many younger embryos directly attacked by wasps hatched immediately. Embryos attacked by wasps hatched as much as a third younger than the peak undisturbed hatching age, and most hatching embryos escaped. Thus hatching is an effective defence against wasp predation, and plasticity in hatching stage allows embryos to balance risks from stage-specific egg and larval predators. Wasp-induced hatching is behaviourally similar to the snake-induced hatching previously described in A. callidryas, but occurs in fewer eggs at a time, congruent with the scale of the risk. Individual embryos hatch in response to wasps, which take single eggs, whereas whole clutches hatch in response to snakes, which consume entire clutches. Embryos of A. callidryas thus respond appropriately to graded variation in mortality risks. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032654 TI - Lack of preference for low-predation-risk habitats in larval damselflies explained by costs of intraspecific interactions. AB - Many studies indicate prey organisms select microhabitats with high structural complexity as a way of reducing risk of predation. We used laboratory experiments to show that damselfly larvae, Ischnura verticalis, suffer higher predation rates from pumpkinseed sunfish in low-density vegetation. However, larvae do not preferentially occupy microhabitats with high vegetation density in either the presence or absence of sunfish; when given a choice, the number of larvae per stem of vegetation was equal across all densities of vegetation. That larvae do not congregate in dense vegetation may reflect costs of aggressive interactions. Results from laboratory experiments indicated larval interactions increase conspicuous behaviours (most notably swimming) and consequently increase fish predation. A subsequent experiment indicated that frequency of larval interactions increases with increased vegetation density when number of larvae/stem is constant. Thus, larval microhabitat selection may reflect a trade off between reduced risk of predation in areas of high vegetation density, caused by reduced fish foraging ability, and increased aggressive larval interactions, due to decreased proximity of larvae. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032655 TI - Capuchins do cooperate: the advantage of an intuitive task. AB - We used a cooperative pulling task to examine proximate aspects of cooperation in captive brown capuchin monkeys, Cebus apella. Specifically, our goal was to determine whether capuchins can learn the contingency between their partner's participation in a task and its successful completion. We examined whether the monkeys visually monitored their partners and adjusted pulling behaviour according to their partner's presence. Results on five same-sex pairs of adults indicate that (1) elimination of visual contact between partners significantly decreased success, (2) subjects glanced at their partners significantly more in cooperative tests than in control tests in which no partner-assistance was needed, and (3) they pulled at significantly higher rates when their partner was present rather than absent. Therefore, in contrast to a previous report by Chalmeau et al. (1997, Animal Behaviour, 54, 1215-1225), cooperating capuchins do seem able to take the role of their partner into account. However, the type of task used may be an important factor affecting the level of coordination achieved. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032656 TI - Subtidal food thieves: interactions of four invertebrate kleptoparasites with the sea star Leptasterias polaris. AB - We used time-lapse underwater video systems in the subtidal zone of the Mingan Islands, eastern Canada, to examine behavioural interactions of four kleptoparasites with the sea star Leptasterias polaris when it was feeding on infaunal (buried) clams. Departures of L. polaris from its prey coincided with interactions with kleptoparasites in 10 out of 10 filmed feeding bouts on the large clam Spisula polynyma, compared with only four out of 10 of filmed bouts on the smaller clam Mya truncata. The sea star's abandoning of the prey was most often caused by interactions with another sea star, Asterias vulgaris, a potential predator. The whelk (a carnivorous snail), Buccinum undatum, and the crabs Hyas araneus and Cancer irroratus, also kleptoparasitized L. polaris, especially when A. vulgaris was present. Comparisons of feeding bouts in the presence and absence of kleptoparasites showed that at least 10.4% of the prey mass captured by L. polaris was lost to kleptoparasites. Simultaneous current meter data showed that the movement of A. vulgaris and B. undatum to sites where L. polaris was feeding on S. polynyma was upstream, suggesting chemodetection of food odours. Crabs in the vicinity may also use visual cues. Leptasterias polaris is the only carnivore that can efficiently extract large clams from the sediments and therefore its foraging may supply a substantial part of the diet of kleptoparasites. Copyright 2000 The Association for the Study of Animal Behaviour. PMID- 11032657 TI - Component processes in task switching. AB - Participants switched between two randomly ordered, two-choice reaction-time (RT) tasks, where an instructional cue preceded the target stimulus and indicated which task to execute. Task-switching cost dissipated passively while the participants waited for the instructional cue in order to know which task to execute (during the Response-Cue Interval). Switching cost was sharply reduced, but not abolished, when the participants actively prepared for the task switch in response to the instructional cue (during the Cue-Target Interval). The preparation for a task switch has shown not to be a by-product of general preparation by phasic alertness or predicting target onset. It is suggested that task-switching cost has at least three components reflecting (1) the passive dissipation of the previous task set, (2) the preparation of the new task set, and (3) a residual component. PMID- 11032658 TI - Expectancy, attention, and time. AB - Seven experiments examine the influence of contextual timing manipulations on prospective time judgments. Subjects judged durations of standard vs comparison time intervals in the context of a preceding induction (context) sequence. In some experiments, the rate of the induction sequence was systematically manipulated relative to the range of to-be-judged standard time intervals; in others, the induction sequence was omitted. Time judgments were strongly influenced by the rate of an induction sequence with best performance occurring when the standard time interval ended as expected, given context rate. An expectancy profile, in the form of an inverted U, indicated that time estimation accuracy declined systematically as a standard interval differed from a context rate. A similar expectancy profile emerged when the context rate was based on a harmonic subdivision (one-half) of an expected standard interval. Results are discussed in terms of various stimulus-based models of prospective time judgments, including those which appeal to attentional periodicities and entrainment. PMID- 11032660 TI - Cellular origins of ultraviolet radiation-induced corneal tumours in the grey, short-tailed South American opossum (Monodelphis domestica). AB - Corneal tumours were induced in almost 100% of grey, short-tailed South American opossums (Monodelphis domestica) exposed three times weekly to ultraviolet radiation (UVR) for periods of a year or more. Five tumours, representing the morphological spectrum of UVR-induced corneal tumours (two fibrosarcomas, one malignant fibrous histiocytoma, one putative haemangiosarcoma, and one squamous cell carcinoma overlying a sarcoma), were assayed immunohistochemically for reactivity with antibodies against the intermediate filaments vimentin, smooth muscle actin (alpha isoform), muscle-specific actins (alpha and gamma isoforms), desmin and cytokeratin, and with antibodies against the vascular endothelial marker von Willebrand factor. The squamous cell carcinoma was cytokeratin positive. Other tumours were cytokeratin-negative and vimentin-positive. Three tumours had scattered individual cells and groups of cells immunoreactive with antibodies against smooth muscle actin and muscle-specific actins; two tumours (a fibrosarcoma and the malignant fibrous histiocytoma) had small numbers of desmin positive cells. The putative haemangiosarcoma contained two populations of neoplastic cells, von Willebrand factor-positive vascular endothelial cells and smooth muscle actin-positive spindle cells. It was concluded (1) that UVR-induced corneal tumours may be composed of cells derived from resident epithelial cells, immigrant vascular endothelial cells, or fibroblast-like cells of unknown origin, and (2) that such tumours may contain more than one neoplastic cell type. PMID- 11032659 TI - Distribution of cells labelled by a monoclonal antibody (A3) against a cloned cell line derived from a rat malignant fibrous histiocytoma. AB - To pursue the histogenesis of malignant fibrous histiocytoma (MFH), of which the cell of origin is still debated, a monoclonal antibody (A3) was produced against a rat MFH-derived cloned cell line (MT-8). Antigen recognized by A3 was around 80 kDa in molecular weight and was seen on the cytoplasmic membrane of MT-8 cells by immunoelectron microscopy. A3 reacted specifically with MT-8 cells, with another rat MFH-derived cell line (MT-9) and with their induced tumours in syngeneic rats, but not with other rat tumours such as fibrosarcoma, histiocytic sarcoma, malignant meningioma, uterine leiomyosarcoma, endometrial stromal sarcoma, mononuclear cell leukaemia and malignant schwannoma. These findings indicate that A3 has a high specificity for rat MFH cells. In fetuses on gestation days 15, 18 and 20 and in postnatal rats aged 1, 4 and 8 days, A3 reacted with primitive mesenchymal cells in visceral organs and around arteries and bronchi, as well as in the lamina propria of intestinal mucosa, renal interstitium, meninges and perineurium. There were no A3-positive connective tissue cells in organs or other sites in adult rats more than 10 weeks old. It is therefore likely that MFH cells share antigens with primitive mesenchymal cells, which may be multipotent for mesenchymal differentiation. The present study suggests that MFH consists of a population of primitive, undifferentiated mesenchymal cells. A3 also immunolabelled endothelial cells of arteries, venules and pulmonary capillaries in fetal, postnatal and adult rats; vascular endothelial cells in chemically induced hepatic and renal lesions also reacted strongly with A3. However, the significance of endothelial immunoreactivity with A3 remains to be elucidated. PMID- 11032661 TI - Immunophenotypical characterization of the lymphocyte infiltrate in caprine pulmonary tuberculosis. AB - A study was made of the immunophenotype of the lymphocytes associated with natural caprine pulmonary tuberculosis at four stages of the disease. Regardless of the stage, CD4(+)and CD8(+)T lymphocytes predominated in lung and mediastinal lymph node lesions, but gamma/delta T and B (IgM(+)) cells were seen only rarely. At the primary complex stage, CD4(+)cells outnumbered CD8(+)cells. At the stage of generalized tuberculosis, however, and still more at the post-primary stage, CD8(+)cells outnumbered CD4(+)cells. At the final stage (tuberculous pneumonia), CD4(+)and CD8(+)cells were present in low but approximately equal numbers. PMID- 11032662 TI - Effect of selenium deficiency on the development of central nervous system lesions in murine listeriosis. AB - The effect of selenium (Se) deficiency, produced by feeding a Se-deficient diet, on the development of central nervous system (CNS) lesions was studied in mice infected with Listeria monocytogenes, administered in drinking water for 1 or 7 days in a daily dose of 10(9)organisms, or for 7 days in a daily dose of 10(7). Se-deficient mice differed from Se-normal controls in developing CNS lesions significantly more frequently. Moreover, regardless of Se status, mice receiving repeated doses of 10(9)organisms differed from those receiving a single 10(9)dose in showing CNS lesions at least twice as often. The majority of animals with CNS lesions showed an inflammatory pattern of rhombencephalitis (17/24), while only two of 24 showed choroiditis-ventriculitis-meningitis; five of 24 animals showed both inflammatory patterns. Listeria monocytogenes antigen was identified within the areas of inflammation by an immunoperoxidase technique. Neuritis of the trigeminal nerve was present in eight animals. The relative lack of pathological changes in the liver and spleen validates this murine model for the study of CNS listeriosis. PMID- 11032663 TI - Spontaneous thyroid-containing teratoma associated with impaired development in the African clawed frog, Xenopus laevis. AB - Teratomas are rare in amphibians and the neoplasm described here, which had a significant thyroid carcinoma component, is the first tumour of this type to be reported in Xenopus laevis. The thyroid component contained moderately to well differentiated acinar glands showing much hyperplasia, dysplasia, and reduced and distorted colloid reservoirs. Cartilaginous, neural, muscular, mesenchymal and gut-like epithelial components were also observed in this ventral mediastinal neoplasm, indicating aberrant proliferation from all three germ layers. This teratoma was only one abnormality in a complex of developmental changes, followed for 28 months, which appeared in a single generation of sibling 2-week-old Xenopus larvae. Two hundred larvae produced by an apparently normal adult pair initially showed ocular defects, including microphthalmia, anophthalmia and tumours projecting near the eyes. During further development up to 28 months, mediastinal tumours developed in nine frogs; these tumours were associated with reduced growth, the frogs reaching only 13-20% of normal weight, and greatly enhanced ventral pigmentation. PMID- 11032664 TI - Demonstration of equine herpesvirus-1 gene expression in the placental trophoblasts of naturally aborted equine fetuses. AB - Equine herpesvirus-1 (EHV-1) infection was demonstrated in the lung tissue of seven aborted fetuses by immunohistochemical labelling and polymerase chain reaction. The placentas of the fetuses were also examined by non-isotopic in-situ hybridization for the EHV-1 glycoprotein B (gB) gene. Positive hybridization signals were observed in the cytoplasm of trophoblasts, especially in microcotyledons, of all seven placentas, and in villous epithelium of the allantochorion of six placentas. Despite the presence of EHV-1 RNA, EHV-1 antigens were not detected in placentas by immunohistochemical examination. The present study represents the first in-vivo demonstration of the EHV-1 gene in equine trophoblasts. The findings suggest direct cell-to-cell spread of EHV-1 from endometrial cells to trophoblasts. PMID- 11032666 TI - Apoptosis in rabbit haemorrhagic disease. AB - Rabbit haemorrhagic disease virus (RHDV) causes an acute hepatitis and disseminated intravascular coagulation. Six rabbits were inoculated experimentally with RHDV to investigate any potential relationship between infection and apoptosis in the liver. Two rabbits were killed at 12 h post inoculation (PI) and two at 24 h PI. The remaining two rabbits died at 30 h and 31 h PI. Immunohistochemical labelling for RHDV antigen-positive cells, TUNEL assay for apoptotic cells, and DNA analysis were performed on samples of liver. The four rabbits that died or were killed 24-31 h PI had acute hepatitis with infiltration of heterophils and necrotic hepatocytes. RHDV antigen-positive cells and apoptotic cells appeared in the centriacinar areas at 12 h PI; subsequently they spread to periacinar areas and increased in number, but the viral antigen positive cells outnumbered apoptotic cells. At 24-31 h PI, few apoptotic cells were recognized in the areas infiltrated with lymphocytes and heterophils. The results suggested an association between RHDV infection and apoptosis of hepatocytes. PMID- 11032665 TI - Immunohistochemical characterization of calf pneumonia produced by the combined endobronchial administration of bovine herpesvirus 1 and Pasteurella haemolytica. AB - Ten calves ("group 4") were inoculated endobronchially with Pasteurella haemolytica 4 days after inoculation with bovine herpesvirus 1 (BHV-1). Four calves (group 3) were similarly inoculated with P. haemolytica alone, and three (group 2) with BHV-1 alone. All group 4 animals showed severe respiratory signs and had bilateral lobar pneumonia; one died 6 days after inoculation with P. haemolytica. Two types of pneumonic lesion were observed. One was characterized by interlobular and interstitial lymphatic thrombosis, fibrinous pleuritis and coagulative necrosis, and the other by necrotizing bronchiolitis with intranuclear inclusion bodies. The former type of lesion was associated with the presence of P. haemolytica antigen and the latter with the presence of BHV-1 antigen. The weight of infection of BHV-1 and P. haemolytica in bronchoalveolar (BAL) fluid was clearly reflected in the immunohistochemical demonstration of the corresponding antigens in BAL fluid cells. Group 4 calves differed from the calves of groups 1-3 in showing 10-1530 times more endotoxin in BAL fluid. These findings suggested that BHV-1 infection partly destroyed the clearance mechanisms of the respiratory tract epithelium and exacerbated the subsequent P. haemolytica infection. PMID- 11032668 TI - Excretory plugs from the choroid plexus in the cerebrospinal fluid of dogs with neurological disease: possible role in the formation of corpora arenacea. AB - Cytological specimens of cerebrospinal fluid (CSF) from 13 (6.5%) of 200 dogs with various neurological diseases contained excretory plugs (EPs) originating from choroid plexus cells (ChPCs). The presence of EPs was not related to age, breed or sex, or to any particular neurological disease. EPs, which ranged from 3 to 25 microm in diameter, had a sharp outline and contained granular material. Many such bodies were also found within ChPCs of dogs without neurological disease, which were examined to determine the origin of EPs. The results suggest that EPs found in the CSF are excretory products originating in a small population of ChPCs, and that when they reach a certain size they are eliminated into the CSF. It is possible that they serve as cores around which corpora arenacea ("brain sand") are subsequently formed. PMID- 11032667 TI - Expression of vascular endothelial growth factor in basal cell tumours and in squamous cell carcinomas of canine skin. AB - The expression of vascular endothelial growth factor (VEGF) was evaluated immunohistochemically in 20 basal cell tumours (BCTs) and 15 squamous cell carcinomas (SCCs) of canine skin. VEGF was identified in all the SCCs and was particularly striking in those occurring on the toe. On the other hand, VEGF was absent in the great majority of BCTs, occurring only in those of the solid type. The results suggest that presence of VEGF is a useful additional criterion for evaluating malignancy and growth potential in tumours of these types. PMID- 11032669 TI - A comparative study of pseudorabies virus (PRV) strains with defects in thymidine kinase and glycoprotein genes. AB - In the course of two experiments, an examination was made of the virulence and neuroinvasiveness for pigs of two pseudorabies virus mutants (strain 6C2TK(-), with a defect in thymidine kinase (TK) function; and strain 6C2TK(-), gI(-)/gE( ), with defects in TK and glycoproteins I and E) and of the wild-type parent strain (86/27V). At various times after intranasal inoculation, pigs were killed and samples of tonsil, lung and different levels of the trigeminal and olfactory nervous pathways were examined by methods that included viral isolation, polymerase chain reaction assay and immunohistochemistry. Both mutant viruses were of reduced virulence, as indicated by no more than moderate clinical signs and lesions, and only sporadic isolation of virus; moreover, unlike the wild-type parent strain, the mutant viruses were not reactivated from the latent state by corticosteroid treatment. In addition, migration of the mutant strains to the central nervous system (olfactory and trigeminal nervous pathways) was reduced as compared with that of the wild-type strain. Thus, mutations in the genes encoding the TK enzyme and the gI/gE complex were associated with reduced virulence, reduced replication in peripheral target tissues, and reduced migration to the olfactory and trigeminal pathways. PMID- 11032670 TI - Expression of beta1 integrin in relation to histological features in normal and neoplastic canine testicles. AB - The expression of the beta1 common chain of the VLA integrin subfamily was evaluated immunohistochemically in a series of five normal and 30 neoplastic canine testicles. The tumours, consisting of seminomas or Sertoli cell tumours, were classified according to WHO criteria as intraductal without signs of invasion, intraductal with signs of invasion, or diffuse. Expression of beta1 integrin decreased progressively from intraductal tumours without signs of invasion, to the diffuse type, in which immunolabelling was generally absent. In a few cases of diffuse neoplasia, groups of neoplastic cells exhibited strong positivity that was not restricted to the basal pole of the cell membrane. These results suggest that the expression of beta1 integrin was related to the histological tumour type, possibly reflecting a specific requirement for a reduction in integrin by neoplastic cells with infiltrative and metastatic potential. PMID- 11032672 TI - Response of air sac mesothelium to expansion of extracellular fluid volume in Gallus domesticus. AB - In this study, scanning electron microscopy was used to examine the effects of hydrothorax on the morphology of the air sac visceral mesothelium of Gallus domesticus. Anaesthetized chickens were subjected to acute hydrodynamic pulmonary oedema induced by expansion of the extracellular fluid volume with an infusion of Ringer's solution equal to 6.5% of body weight. Tissue samples from the visceral surface of the abdominal air sacs near their ostia were obtained and fixed after death induced by anaesthetic overdose. These were compared with similar samples from control "non-volume-loaded" birds. The air sac visceral mesothelium of the volume-loaded animals presented an increased density of bulbous or swollen microvilli. These deformations were similar to changes reported in the visceral pleura of mammals subjected to hydrothorax, suggesting a commonality with regard to the role of these mesothelia in liquid clearance during pulmonary oedema. PMID- 11032671 TI - Chlamydophila abortus (Chlamydia psittaci serotype 1) clearance is associated with the early recruitment of neutrophils and CD8(+)T cells in a mouse model. AB - The immune mechanisms in response to Chlamydophila abortus (Chlamydia psittaci serotype 1) infection were studied in C57BL/6 and CBA mice. The infection was monitored and the following aspects of the immune response were evaluated: the nature of the leucocyte infiltrate in the liver, the percentages of polymorphonuclear neutrophils (PMNs), macrophages and lymphocytes in the spleen, and the concentrations of cytokines in serum. In addition, the serum concentrations of IgG1 and IgG2a were determined. Both mouse strains showed a Th1 like immune response, with high concentrations of IFN-gamma and minimal levels of IL-4; however, C57 mice differed from CBA mice in showing milder clinical signs and earlier resolution of infection. The greater ability of C57 mice than CBA mice to eliminate chlamydophilae was related to the establishment of an earlier innate immunity, based on a more pronounced PMN response, and on a greater presence of CD8(+)T cells. PMID- 11032673 TI - Immunohistochemical detection of virulence-associated Rhodococcus equi antigens in pulmonary and intestinal lesions in horses. AB - Two horses with Rhodococcus equi infection were examined post mortem by an immunohistochemical method (peroxidase-antiperoxidase; PAP) with a monoclonal antibody (Mab 10G5) to the 15-17 kDa antigen of R. equi. One of the horses was also examined bacteriologically, R. equi being isolated in culture. Immunolabelling with this Mab was marked and widespread. On the other hand, the immunohistochemical reactivity of infected macrophages with a polyclonal antibody specific for lysozyme was slight. Thus, Mab 10G5 would appear to be a useful diagnostic reagent in R. equi infection, with or without cultural confirmation. PMID- 11032674 TI - An outbreak of pulmonary mycosis in respiratory burst-deficient (gp91(phox-/ ))Mice with concurrent acidophilic macrophage pneumonia. AB - An outbreak of pulmonary abscessation due to the common environmental fungus Paecilomyces variotii occurred in a colony of gp91(phox-/-)mice, which lack functional NADPH oxidase, the enzyme complex responsible for generating the respiratory burst in phagocytes. Parenchyma surrounding the abscesses showed intense acidophilic macrophage pneumonia, and similar but much milder lesions were present in each of 24 gp91(phox-/-)mice free from other pulmonary lesions. These findings indicate a high prevalence of acidophilic macrophage pneumonia in gp91(phox-/-)mice, or at least in this particular stock. The presence of this lesion may complicate studies in which phagocyte-dependent pulmonary defence plays a significant role. PMID- 11032675 TI - Systemic infection by Pasteurella canis biotype 1 in newborn puppies. AB - Pasteurella canis biotype 1, usually associated with the oral cavity of dogs and cats, or with human wound infections following dog bites, was isolated from newborn puppies with a fatal systemic infection. The identity of P. canis was confirmed by arbitrarily primed polymerase chain reaction and the organism was susceptible to all the penicillins, cephalosporins, tetracyclines and fluoroquinolones tested and to most of the aminoglycosides tested. This represents the first report of systemic pasteurellosis caused by P. canis in dogs. PMID- 11032676 TI - Pathological findings in two fin whales (Balaenoptera physalus) with evidence of morbillivirus infection. AB - Two immature female fin whales stranded on the Belgian and French coastlines, were examined post mortem. The main gross findings were massive parasitic infestation, associated with a large thrombus in one whale, and severe emaciation. Microscopical investigations revealed multinucleated syncytia with large intranuclear inclusion bodies in various tissues, and positive immunolabelling for morbillivirus antigens. Other evidence of morbillivirus infection was provided by the demonstration of specific viral structures in syncytia and in cell cultures, and the detection of neutralizing antibodies to canine distemper virus. To the authors>> knowledge, this is the first firm report of morbillivirus infection in baleen whales. PMID- 11032678 TI - Diffuse coronary arteriodysplasia in a Holstein-Friesian cow. AB - The coronary arteries of a 6-year-old Holstein-Friesian cow showed dysplastic lesions characterized by the following vascular changes: irregular, aneurysmal dilation; intimal fibrosis associated with myointimal cells; aberrant internal elastic lamina; oedema and mast cell infiltrates; and variation in the thickness of the media, including tortuosity with irregular dilation of the lumen of the artery and a plexiform structure formed from the right coronary artery. PMID- 11032677 TI - Antioxidant status in a squirrel monkey (Saimiri sciureus) with chronic pancreatitis and degenerative myopathy. AB - A squirrel monkey (Saimiri sciureus) presented with wasting, vomiting and diarrhoea. Haematology revealed elevation of creatinine phosphokinase, lactic dehydrogenase, alanine aminotransferase, amylase and lipase, together with azotaemia and hypoalbuminaemia. Prominent findings were chronic pancreatitis with acinar and ductal plugs, granulomatous and necrotizing peripancreatic steatitis, degenerative myopathy, testicular atrophy, candidiasis and bacterial necrotizing glossitis. Antioxidant analyses revealed low concentrations of serum vitamin E (and apparently A), hepatic selenium and hair zinc. Pancreatitis may have caused malabsorption and maldigestion, associated with deficiency of multiple antioxidants. PMID- 11032679 TI - Multiple endocrine neoplasia in a dog. AB - Multiple endocrine neoplasms, including an insulinoma, bilateral adrenocortical adenocarcinomas and an aortic paraganglioma, were diagnosed after euthanasia in a 12-year-old spayed female dog of mixed breed with a history of progressive anorexia, vomiting, diarrhoea, weight loss, polyuria and polydipsia, regenerative anaemia and hypoglycaemia. The clinical, gross pathological, microscopical, immunohistochemical and ultrastructural findings were consistent with the human syndrome of multiple endocrine neoplasia. PMID- 11032680 TI - Renal microcystic tubular lesions in two 1Year-old dogs - an early sign of hereditary renal cystadenocarcinoma? AB - As a part of a study of early renal changes in renal cystadenocarcinoma (RC), a 5 year-old German shepherd dog and two 1-year-old German shepherd mixed-breed dogs were examined. All three animals had bilateral, microscopic renal cysts, and the 5-year-old dog also had RC. Microscopical examination showed papillary hyperplastic tubular epithelial cells lining the inner wall of the renal cysts in all dogs. These cells showed strong reactivity with a monoclonal antibody against a broad-spectrum type of cytokeratin. The dam of the young dogs had suffered from autosomal dominant inherited RC and nodular dermatofibrosis (ND) syndrome. It is likely that the microscopic renal cystic lesions seen in the young dogs represented an early renal change in the RC/ND syndrome. This suggests that the diagnosis of RC can be made on suspected carriers by microscopical examination of renal biopsies as early as 1 year of age, i.e., before the dogs are used for breeding. PMID- 11032681 TI - Papillary meningioma in the cerebellum of a cat. AB - An unusual papillary meningioma in a cat with progressive cerebellar ataxia is described. Computed tomography revealed the presence of a primary tumour. Microscopically, the tumour was seen to arise from the cerebellar pia mater, forming perivascular structures composed of a single layer of cuboidal to columnar epithelial cells. By immunolabelling, the neoplastic cells appeared positive for cytokeratin, vimentin and S-100 protein. Ultrastructural examination, together with the immunocytochemical findings, indicated the mesenchymal and epithelial nature of the tumour. PMID- 11032684 TI - Preface. PMID- 11032682 TI - Desquamative form of cryptogenic fibrosing alveolitis in a cat. AB - Bronchopulmonary disease is not uncommon in cats, many cases falling into the categories of chronic bronchitis and the "feline asthma syndrome". We report a case of chronic bronchopulmonary disease in an adult cat, which was initially diagnosed as chronic bronchitis. Failure to respond to appropriate therapy led to euthanasia. At necropsy, the lungs exhibited multifocal areas of consolidation, especially at the periphery of the diaphragmatic lobes. Histopathological examination revealed a striking variability of lesions, with interstitial fibrosis and intra-alveolar accumulations of macrophages in addition to alveolar epithelialization and smooth muscle hyperplasia. These changes were consistent with those described for the desquamative form of cryptogenic fibrosing alveolitis in human beings. PMID- 11032685 TI - There is a non-linear relationship between mortality and blood pressure. PMID- 11032686 TI - Blood pressure and long-term coronary heart disease mortality in the Seven Countries study: implications for clinical practice and public health. PMID- 11032687 TI - Percutaneous balloon valvuloplasty - the first line treatment for mitral stenosis and restenosis. PMID- 11032688 TI - Homograft - the optimal aortic valve substitute? PMID- 11032689 TI - White coat effect. Innocuous or adverse phenomenon? PMID- 11032690 TI - Vascular endothelial growth factor and coronary artery surgery: implications for improved endothelial protection! PMID- 11032691 TI - Percutaneous mitral balloon commissurotomy: a useful and necessary treatment for the western population. PMID- 11032692 TI - Hypertensive heart disease. A complex syndrome or a hypertensive 'cardiomyopathy'? PMID- 11032693 TI - Usefulness and limitations of dobutamine-atropine stress echocardiography for the diagnosis of coronary artery disease in patients with left bundle branch block. A multicentre study. AB - BACKGROUND: Patients with left bundle branch block exhibit abnormal septal motion which may limit the interpretation of stress echocardiograms. This study sought to assess the diagnostic value of dobutamine-atropine stress echocardiography in left bundle branch block patients. METHODS AND RESULTS: Sixty-four left bundle branch block patients (mean age 59 years, 24 men) with suspected coronary artery disease underwent dobutamine-atropine stress echocardiography and coronary arteriography. Myocardial ischaemia was defined as new or worsening wall thickening abnormalities. Coronary artery disease was quantitatively defined as a diameter stenosis >/=50% in a major epicardial artery. Rest septal motion was normal (apart from the early systolic septal notch) in 34 patients (53%) and abnormal in 30 patients (47%). Rest septal thickening was normal in 32 patients (50%) and abnormal in 32 patients (50%). All seven patients with a QRS duration >/=160 ms and an abnormal QRS axis had abnormal rest septal motion and thickening. Inter-observer agreement for ischaemia was 88%. In all but one patient disagreement was in the septum. For the anterior and posterior circulation, respectively, sensitivity was 60% (9/15) and 67% (8/12), specificity was 94% (46/49) and 98% (51/52), and accuracy was 86% (55/64) and 92% (59/64). Sensitivity for the anterior circulation tended to be better in patients with normal rest septal thickening (83% vs 44%). CONCLUSIONS: Dobutamine-atropine stress echocardiography has excellent diagnostic specificity in left bundle branch block patients with suspected coronary artery disease. In patients with abnormal rest septal thickening, however, dobutamine-atropine stress echocardiography may lack good sensitivity for detection of coronary artery disease in the anterior circulation. Left bundle branch block patients who potentially most benefit from dobutamine-atropine stress echocardiography may initially be selected by their resting electrocardiogram. PMID- 11032694 TI - Assessment of coronary calcification by electron-beam computed tomography in symptomatic patients with normal, abnormal or equivocal exercise stress test. AB - AIMS: Exercise stress testing is often used as the initial non-invasive diagnostic test in symptomatic patients with suspected obstructive coronary artery disease. Positive standard ECG criteria are quite specific for obstructive coronary artery disease, but there may be a substantial number of false negative tests, including patients with severe coronary artery disease. Also, exercise stress tests frequently yield equivocal results. Instead of detecting the functional consequences, electron-beam computed tomography visualized atherosclerotic plaque disease directly, but its relationship to functional testing has not been clearly delineated. It was the aim of the current study to examine electron-beam computed tomography for the identification of obstructive coronary artery disease in patients with a normal, abnormal, or equivocal exercise stress test. METHODS AND RESULTS: Symptomatic patients referred for coronary angiography were prospectively included in a consecutive manner if they had no prior diagnosis of coronary artery disease and an unremarkable resting ECG. All patients underwent both exercise stress test and electron-beam computed tomography on the day before coronary angiography. Standard protocols and ECG criteria to diagnose inducible ischaemia were used for the exercise stress test. The electron-beam computed tomography-derived total calcium score was computed according to standard Agatston criteria. Of the 323 patients (mean age, 55+/-11 years; 77% male), 179 (55%) had obstructive coronary artery disease, defined angiographically as luminal diameter narrowing >/=50%. A normal exercise stress test was documented in 105 patients (32.5%), an abnormal exercise stress test ('diagnostic for ischaemia') in 113 (35%), and an equivocal exercise stress test ('inadequate exercise or non-diagnostic ECG-changes') in 105 (32.5%). Multivariate analysis indicated that exercise stress test and electron-beam computed tomography yielded independent information for predicting obstructive coronary artery disease. Sensitivity, specificity and overall accuracy of the exercise stress test were 71%, 75% and 73%, respectively, if equivocal tests were not included, and 50%, 84% and 65% if they were included. Irrespective of the cutpoint regarded as 'positive', the overall accuracy of the electron-beam computed tomography-derived calcium score remained approximately 80% in patients with a normal, abnormal or equivocal exercise stress test. In patients with an equivocal and - to a lesser degree - with a normal exercise stress test, electron beam computed tomography was able to significantly improve classification regarding obstructive coronary artery disease. Electron-beam computed tomography added no incremental predictive value in patients with an abnormal exercise stress test. CONCLUSION: In patients who are judged to have an intermediate post test probability of disease after exercise stress test, electron-beam computed tomography scanning may be a meaningful strategy for further stratification regarding the likelihood of obstructive coronary artery disease. PMID- 11032695 TI - Immediate and mid-term results of repeat percutaneous mitral commissurotomy for restenosis following earlier percutaneous mitral commissurotomy. AB - AIMS: This study assessed the results of repeat percutaneous mitral commissurotomy for mitral restenosis following a first procedure. METHODS AND RESULTS: Repeat balloon commissurotomy was performed in 53 patients who had symptomatic restenosis a mean of 6+/-2 years (2-11) after a successful first procedure; seven patients had mildly calcified valves. All patients had restenosis with a fusion of both commissures as assessed by echocardiography. A double-balloon was used in one case and the Inoue technique in 52. Complications were stroke in one patient and severe mitral regurgitation (Sellers grade 3) in two. Valve area increased from 1.03+/-0.22 to 1.82+/-0.21 cm(2)(P<0.0001) as assessed by planimetry. Good immediate results, defined as valve area >/=1.5 cm(2)with no regurgitation >2/4, were obtained in 48 patients (91%). The 5-year survival rate without operation and in NYHA class I or II was 69+/-11% in the whole population, and 76+/-11% in the 48 patients who had had good immediate results. CONCLUSION: This study suggests that repeat balloon commissurotomy is a valid treatment for symptomatic restenosis after a first successful procedure. It gives good results in patients selected on the basis of favourable characteristics and the echocardiographic analysis of the mechanism of restenosis. PMID- 11032696 TI - Inoue balloon mitral valvuloplasty: long-term clinical and echocardiographic follow-up of a predominantly unfavourable population. AB - AIMS: To assess long-term outcome in a typical Western population of predominantly unfavourable patients undergoing Inoue balloon mitral valvuloplasty. Outcome amongst patients has only been undertaken in the medium term. Long-term echocardiographic data in particular are scarce. METHODS: Inoue mitral valvuloplasty was attempted in 106 patients. There were six technical failures; the procedure was therefore completed in 100 patients, who underwent annual clinical and echocardiographic follow-up. RESULTS: Patients were aged 63.5+/-10. 3 years. 82% were female. Unfavourable characteristics included age >65 (52%), NYHA class III or IV (87%), >/=1 significant co-morbidity (63%), atrial fibrillation (82%), previous surgical commissurotomy (25%) and echocardiographic score >8 (59%, mean 8.9+/-2.1). Mitral valve area increased from 0.98+/-0.23 to 1.54+/-0.31 cm(2). There were three major complications. Post procedure, symptoms improved in 88% of patients. Haemodynamic success (mitral valve area increase >50%, final mitral valve area >1.5 cm(2), mitral regurgitation 50% gain in mitral valve area, mitral valve area <1.5cm (2)) was 98%, 92% and 75% at 1, 3 and 6 years. Pre-procedural predictors of event-free survival were male sex, absence of co-morbidities, lower echocardiographic score and smaller left atrial diameter. CONCLUSIONS: In a Western population with predominantly unfavourable characteristics for mitral valvuloplasty, long-term outcome post-procedure is reasonable. A moderate increase in mitral valve area can be achieved at low procedural risk, and the subsequent rate of restenosis is low. Nonetheless, 6 years after the procedure, half of the patients will have required further intervention or died. For fitter patients willing to accept significant operative risk, mitral valve replacement remains a valuable alternative. PMID- 11032697 TI - Long-term results after aortic valve replacement in patients with congestive heart failure. Homografts vs prosthetic valves. AB - OBJECTIVES: The aim of this study was to assess the influence of valve substitute (homograft vs prosthetic valve) on the long-term survival and late valve-related complication rates following aortic valve replacement in patients with aortic valve disease and congestive heart failure. BACKGROUND: The effect of choice of valve substitute on outcome after aortic valve replacement in patients with pre operative heart failure is unknown. The superior haemodynamic profile of homografts may be of particular benefit. METHODS: We retrospectively analysed pre operative, operative and follow-up data on 518 adults in functional classes III and IV, who, over the 25 years 1969-1993, had their initial aortic valve replacement at Harefield hospital. Follow-up conducted during 1996 to April 1997 and totalling 4439 patient-years was 96.1% complete. Using multivariate analysis, independent risk factors for different complications and mortality were defined. RESULTS: Overall 5-, 10- and 20-year survival was 80+/-2%, 62+/-2% and 30+/-3%, respectively, with no significant difference between valve types. However, homografts (n=381) independently reduced the rate of serious complications and cardiac death, whereas mechanical valves were an independent adverse risk factor for late mortality. The rates of anticoagulant-related bleeding and thromboembolism were increased by mechanical valves, whereas primary tissue failure was the main complication of homografts. CONCLUSIONS: Long-term outcome of homograft aortic valve replacement in patients with congestive heart failure is acceptable, with a reduced rate of serious complications and cardiac death. Further improvements would be expected if the rate of primary tissue failure could be reduced. PMID- 11032698 TI - Vascular endothelial growth factor release following coronary artery bypass surgery: extracorporeal circulation versus 'beating heart' surgery. AB - AIMS: The aim of this study was to examine the circulating levels of vascular endothelial growth factor, following coronary artery bypass graft surgery performed using both standard cardiopulmonary bypass or the 'octopus technique' on the beating heart. BACKGROUND: Vascular endothelial growth factor has a number of effects that are beneficial in the setting of coronary artery bypass graft surgery including cardioprotection, potent angiogenic activity and amelioration of intimal hyperplasia. Hypoxia is a powerful stimulator of vascular endothelial growth factor expression yet the ability of ischaemia, occurring during coronary artery bypass graft surgery, to induce vascular endothelial growth factor production is unknown. METHODS AND RESULTS: Serum vascular endothelial growth factor levels were determined in patients undergoing coronary artery bypass graft surgery with standard cardiopulmonary bypass (CPB-CABG group; n=20), with off pump coronary artery bypass; (OP-CABG; n=12) and in patients undergoing non cardiac major surgery (n=6). The effect of hypoxia on vascular endothelial growth factor release by neonatal rat cardiac myocytes in vitro was studied. In the CPB CABG group vascular endothelial growth factor levels were significantly increased to 78.5+/-39.3 and 110.5+/-16.3 pg. microl(-1)8 and 24 h post-operatively, declining to 14.9+/-9.9 pg. microl(-1)by 48 h to pre-operative values (14.4+/-8.6 pg. microl(-1)). Significantly higher vascular endothelial growth factor levels were also present in the OP-CABG group 3, 6 and 24 h post-operatively (levels 136. 6+/-29.3, 143+/-26.12 pg. microl(-1)and 93.5+/-20.1 pg. microl(-1), respectively). However, non-cardiac major surgery did not result in elevated vascular endothelial growth factor levels post-operatively (46.36+/-9.76 vs pre surgery levels of 26.84+/-6.1 pg. microl(-1)). Either 15 min or 3 h of hypoxia stimulated vascular endothelial growth factor release from neonatal rat cardiac myocytes in vitro. Twenty-four and 48 h post hypoxia, levels of vascular endothelial growth factor were significantly elevated by approximately 17.5- and 48.5-fold respectively. CONCLUSIONS: These data demonstrate myocardial ischaemia secondary to CPB-CABG and OP-CABG to be a potent stimulator of vascular endothelial growth factor production, which may have implications for graft endothelialization and cardiovascular haemodynamics post-operatively. PMID- 11032700 TI - ESC news and appointments PMID- 11032699 TI - White coat effect, blood pressure and mortality in men: prospective cohort study. AB - BACKGROUND: Because long-term follow-up studies, which also included normotensive controls, have been lacking, the clinical significance of 'white coat' effect and of 'white coat' hypertension has remained controversial. METHODS AND RESULTS: Twenty-one-year prospective data was gathered in 536 men with cardiovascular risk factors at baseline. Blood pressure was measured both by a nurse and by a physician and 'white coat effect' was defined as the difference between the two measurements (physician minus nurse). In addition, four blood pressure groups were categorized: normotensive (n=259), white coat hypertensive (n=18), mildly hypertensive (n=150) and persistently hypertensive (n=109). Comparison of these groups at baseline showed that men with white coat hypertension had higher levels of metabolic risk factors. Sixty-eight men died during follow-up. The men with a white coat effect >30 mmHg (n=37) had significantly higher mortality than other men (relative risk 2.2, 95% confidence interval 1.1-4.2). Mortality was significantly higher in the white coat hypertensive group (33.3%) than in the normotensive group (9.5%, P=0.0005 between groups). Relative risk adjusted for baseline risk factors in the white coat hypertensive group was 3.3 (1.2-7.6) compared with the normotensive group. The development of drug-treated hypertension was also more common (27.8% vs 13.4% in the normotensive group, P<0.0001 between groups). CONCLUSION: The results suggest that white coat hypertension or a large white coat effect is not an innocent phenomenon. It tends to co-exist with metabolic risk factors and predicts total and cardiovascular mortality during long-term follow-up. PMID- 11032701 TI - Differences in the effects of 20 K- and 22 K-hGH on water retention in rats. AB - Antidiuretic actions induced by two growth hormone (GH) isoforms (20 K- and 22 K hGH; 0.2 and 2.0 mg/kg) were evaluated in rats, as fluid retention may cause oedema, one of the adverse effects of GH. Both GH isoforms (2.0 mg/kg) suppressed urine excretion in hypophysectomized rats (P< 0.01), but only the 22 K-hGH isoform (2.0 mg/kg) suppressed urine excretion in intact rats (P< 0.01). In addition, prolactin (PRL) suppressed urine excretion in intact rats (P< 0.05). In conclusion, 20 K-hGH has less potency in causing urine retention than 22 K-hGH and since 20 K-hGH is missing 15 amino acids found in 22 K-hGH, these amino acids may be important for the antidiuretic action of GH. Since prolactin suppressed urine excretion, a part of the antidiuretic action of GH may be related to PRL-R activation. PMID- 11032702 TI - The pharmacokinetics, pharmacodynamics, safety and tolerability of a single dose of NN703, a novel orally active growth hormone secretagogue in healthy male volunteers. AB - The aim of the present study was to investigate the pharmacodynamics, pharmacokinetics, safety and tolerability of a single dose of NN703 (tabimorelin), a growth hormone secretagogue in healthy male subjects. The study design was double blind, randomized and placebo-controlled, with eight escalating dose levels (0.05-12 mg/kg bodyweight (BW)). NN703 was well tolerated by the subjects. The GH area under the curve (AUC) (0-24 h) was significantly higher when compared to placebo for the three highest dose levels (3.0 mg/kg: P = 0.027, 6.0 mg/kg: P = 0.0023, 12 mg/kg: P< 0.0001), and for GH maximal concentration C(max)the four highest dose levels were also significantly higher when compared to placebo (1.5 mg/kg: P = 0.04, 3.0 mg/kg: P = 0.0143, 6.0 mg/kg: P = 0.0053, 12 mg/kg: P = 0.0007). Furthermore, there was a significant increase in IGF-1 levels when compared to placebo for the 6.0 and 12.0 mg/kg BW dose levels (P< 0. 0001). Statistical analysis comparing the AUC (0-24 h) of the NN703 (four highest dose levels) and placebo-treated groups showed no significant increases following NN703 for ACTH, LH, FSH, TSH, prolactin, and cortisol, however, subtle individual changes were noted in ACTH, cortisol and prolactin at doses above 3.0 mg/kg. In conclusion, NN703 is a promising potential candidate for treatment of GH deficiency/insufficiency. PMID- 11032703 TI - Comparison of the somatogenic action of 20 kDa- and 22 kDa-human growth hormones in spontaneous dwarf rats. AB - The somatogenic action of the 20 kilodalton human growth hormone (20 K) was studied using the spontaneous dwarf rat (SDR), which has an isolated GH deficiency. Saline or 2.5 microg, 10 microg, or 100 microg/rat/day of recombinant 20 K or 22 K was administered to prepubertal male and female SDRs for 10 days. Their body weights, serum IGF-I, glucose and insulin were measured, and their body composition was determined. Body weights and serum IGF-I increased dose dependently in both the 20 K- and 22 K-treated groups. There was no significant difference in body weights and serum IGF-I between the 20 K- and 22 K-treated groups except at the dose of 100 microg/rat, in which the IGF-I concentrations were higher in the 22 K-treated male SDRs (P< 0.05: 20 K vs 22 K). Blood glucose was not significantly different between the Spague-Dawley (SD) normal rats and the SDR control groups; however, serum insulin levels of the SDR were higher than those of the SD control group (P< 0.05). Additionally, there was a tendency for serum insulin and glucose levels to increase following 22 K treatment, but the differences were not significant. The percentage of body fat decreased with hGH treatment in both groups (P< 0.01: GH 10, 100 microg/rat group vs SDR control group), however, no significant differences were observed in body composition between the 20 K and 22 K treatment groups. In summary, the 20 K-hGH showed almost the same somatogenic activity as the 22 K-hGH in prepubertal male and female SDRs. PMID- 11032704 TI - Reversible dimerization of 20 kilodalton human growth hormone (hGH). AB - A noncovalent dimer of the 22 kilodalton human growth hormone (22 K-hGH) is known to have diminished somatogenic activity compared with monomeric 22 K-hGH. In the present study, we examined the biological activity and physicochemical behaviour of a noncovalent dimer of the 20 kilodalton human growth hormone (20 K-hGH), an isoform of 22 K-hGH. Analysis of the equilibrium between monomeric and associated forms revealed that the associated 20 K-hGH was present in the dimeric form in aqueous solution. The kinetics of dimerization in rat plasma followed the theory of dissociation-association equilibrium, and more than 99% of 20 K-hGH molecules existed as a monomer in the equilibrium state at the physiological hGH concentration. Analysis of the pharmacokinetics showed that the ratio of the administrated dimer in rat circulation decreased from 43% to less than 4% in 2 h. A preparation of noncovalent dimeric 20 K-hGH had essentially the same degree of biological potency as that of a monomer in both in vitro and in vivo bioassays. In conclusion, dimerization of 20 K-hGH is reversible both in vitro and in vivo and a noncovalent dimer can function as a pharmaceutically active component of a 20 K-hGH preparation, in contrast to a 22 K-hGH preparation. PMID- 11032705 TI - A purified bovine serum albumin preparation contains an insulin-like growth factor (IGF) binding protein-3 fragment that forms ternary complexes selectively with IGF-II and the acid-labile subunit. AB - Among the six insulin-like growth factor binding proteins (IGFBP), only IGFBP-3 and IGFBP-5 form ternary complexes with IGFs and the acid-labile sunbunit (ALS). In a commercial, highly-purified BSA preparation, ternary complex formation was detected using radio-labeled IGF-II and human serum-derived ALS, with precipitation by ALS antiserum. In contrast, no complexes with radio-labeled IGF I were detected under the same conditions. Size-fractionation of the BSA on Superose-12 showed the peak of ternary complex forming activity at approximately 30 kDa. To purify the active factor, a solution of the BSA was pumped onto a [Gly(1)]IGF-II affinity column, and eluted fractions were lyophilized and applied to a C18 HPLC column. The eluted fractions showing ternary complex forming activity maintained a preference for IGF-II in forming ternary complexes and a slight preference in forming binary complexes with IGF-II rather than IGF-I. By silver staining after non-reducing SDS-PAGE, the peak activity in the HPLC-eluted fractions appeared as 30 kDa and 21-24 kDa bands. Amino-terminal sequencing of this peak activity revealed bovine IGFBP-3. These results demonstrate that amino terminal proteolyzed bovine IGFBP-3 is present in a highly purified BSA preparation. In contrast to intact human IGFBP-3 and IGFBP-5, this form of bovine IGFBP-3 forms ternary complexes preferentially with IGF-II rather than IGF-I. PMID- 11032706 TI - Effect of IGFBP-derived peptides on incorporation of(35)SO(4)into proteoglycans. AB - 18 amino acid peptides from the C-terminal region of IGFBP-3, -5 (P3, P5), increased the incorporation of(35)SO(4)into proteoglycans in endothelial cells with greater stimulation in large vessel than microvessel cells. The homologous region of IGFBP-6 (P6) also stimulated sulfate uptake, but less potently than P3 and P5. P6 variants were synthesized with one or two amino acids changed to the basic amino acid in the equivalent position of P3. The P6 variants with one additional basic amino acid behaved similarly to P6. The P6 mutant with two altered amino acids was equipotent to P3. P3F, a scrambled version of P3 was less effective than P3. P3, P5, P6, P3F and all P6 variants all stimulated glucose uptake, which occurred only in microvessel cells. P1, P2, P4, and equimolar intact IGFBP-3 stimulated neither glucose uptake nor sulfate incorporation. Thus, C-terminal basic portions of IGFBP-3, -5 and -6 alter two specific functions of endothelial cells with sufficient differences to suggest mediation by distinct mechanisms. PMID- 11032707 TI - Changes in lipoprotein (a) levels measured by six kit methods during growth hormone treatment of growth hormone deficient adults. PMID- 11032708 TI - Changes in lipoprotein (a) levels measured by six kit methods during growth hormone treatment of growth hormone deficient adults PMID- 11032709 TI - The newborn skin PMID- 11032710 TI - The epidermal barrier. AB - A mature epidermis is an effective barrier which prevents dehydration from the loss of body water, poisoning from the absorption of noxious substances, and systemic infection from invading surface microorganisms. The epidermal barrier resides within the most superficial layer of the skin, the stratum corneum. In utero the fetus has no need for a skin barrier, so the stratum corneum does not start to develop until around 24 weeks' gestation. After 24 weeks there is a steady increase in the number of epidermal cell layers and in epidermal thickness, although it is not until around 34 weeks' gestation that a well defined stratum corneum has completely developed. A weak epidermal barrier is, therefore, present is very preterm infants (<30 weeks' gestation) during the first 2-3 weeks of life and if the skin is damaged by trauma or disease. PMID- 11032711 TI - Clinical consequences of an immature barrier. AB - The epidermal barrier is well developed in the term infant. It is poorly developed in the most immature infants, leading to three major effects: high transepidermal water loss (TEWL), percutaneous absorption, and trauma. A high TEWL leads to poor temperature control and difficulty in fluid balance: it can and should be reduced by manipulating the ambient humidity or by covering the skin. Topical antiseptics should be used sparingly and with care to prevent toxic damage from absorption. The chances of trauma can be reduced by careful choice of monitoring probes and adhesives. Since the epidermal barrier develops rapidly in the postnatal period, these effects only cause problems in the first week or two of life. PMID- 11032712 TI - Adhesives and emollients in the preterm infant. AB - This chapter focuses on recent advances in preterm infant skin care related to skin adhesion and skin emolliency. Different types of adhesives and hydrating agents are reviewed. Clinical applications are best guided by understanding the biology of epidermal barrier development. The role of xeric stress in accelerating formation of the stratum corneum is discussed along with the effects of occlusive agents and emollients on wound healing and epidermal barrier repair. The principles of skin moisturization are introduced. The concept is advanced that programmatic changes in skin adhesion and water handling occur during the normal ontogeny of superficial biofilms (sebum, sweat, acid mantle). PMID- 11032713 TI - The dermis. AB - All the dermal structures are less well developed in the newborn than in the older infant or child, but there are few important consequences of this. Sweating in response to a thermal stimulus occurs at birth in the term infant and can be detected in most preterm infants from 2 weeks of age. It is poorly developed though. Emotional (palmar/plantar) sweating is present from birth in term infants only. Skin blood flow can be regulated in term and preterm infants, and is often measured indirectly as a temperature gradient. Such a gradient is temperature as well as illness dependent which limits its use as a clinical tool. Sensory nerve endings are readily stimulated in the most immature infants. Finally, damage to the skin in the newborn period commonly leads to scarring, although this usually improves with time. PMID- 11032714 TI - Common skin conditions in the newborn. AB - Common or relatively common skin conditions which may be seen in the first month of life are considered. These include benign transient lesions, developmental and pigmentary abnormalities, vascular disorders and infections. PMID- 11032715 TI - Genetic skin disorders. AB - Neonatologists do not require a detailed knowledge of all genetic skin disorders but need to recognize one if they see it. The unique accessibility of the skin makes it possible to observe the physical signs and deduce the child's immediate needs from first principles. The morphological classification given here will help the nondermatologist establish a clinical diagnosis. Tremendous advances over the last 10 years in understanding the molecular basis of skin disease make it possible, in many cases, to confirm the diagnosis and to counsel the family accurately. PMID- 11032716 TI - Management of severe blistering disorders. AB - Sufferers of the genetically determined blistering skin condition epidermolysis bullosa require specialized management throughout life. This management must begin shortly after birth in order to minimize additional skin damage, some of which may lead to permanent disability. Even the gentle handling practised routinely in neonatal units will result in painful blistering and loss of skin in the severely affected infant. Procedures such as cannulation, screening and monitoring must be modified if unnecessary trauma is to be avoided. PMID- 11032717 TI - Management of large birthmarks. AB - Birthmarks are commonly seen in neonates. Most are insignificant incidental findings and straightforward in their diagnosis and management. Others are of more serious concern and may require intervention for medical or cosmetic reasons. They may also be an indication of abnormalities in other organ systems. This chapter will focus on the diagnosis and management of large vascular and pigmented birthmarks. A multidisciplinary team approach involving paediatricians, dermatologists, plastic surgeons and other specialists is often required in the management of these children. PMID- 11032718 TI - Disruption of aldose reductase gene (Akr1b1) causes defect in urinary concentrating ability and divalent cation homeostasis. AB - Aldose reductase (AKR1B1) is the first enzyme in the polyol pathway through which glucose is converted to sorbitol, and has been implicated in the etiology of diabetic complications. However, its physiological role is still not well understood. In the kidney, AKR1B1 is quite abundant in the collecting tubule cells and thought to provide protection against hypertonic environment. We report here that the mice lacking AKR1B1 showed hypercalciuria, hypercalcemia, hypermagnesemia, and reduced ability to concentrate urine, suggesting a new physiological role of AKR1B1 in divalent cation homeostasis. PMID- 11032719 TI - Thyroid oxidase (THOX2) gene expression in the rat thyroid cell line FRTL-5. AB - A cDNA encoding an NADPH oxidase flavoprotein was isolated from the rat thyroid gland. The predicted 1517-residue polypeptide was 82.5% identical to the human THOX2/DUOX2 and 74% similar to THOX1/DUOX1. Rat THOX2 lacks a stretch of 30 residues, corresponding to one exon in the human gene sequence. THOX2 mRNA was found to be expressed in cultured FRTL-5 cells. The level of THOX2 mRNA was increased by cAMP in these cells and it was decreased in the thyroids of rats treated with the antithyroid drug methimazole, unlike the TPO and NIS mRNAs. Since it was found in the intestine, duodenum, and colon, in addition to thyroid, we suggest that it be called LNOX, the new family of long homologs of NOX flavoproteins rather than THOX and/or DUOX. PMID- 11032720 TI - Enhancement of TNF-alpha-induced apoptosis by immobilized arginine-glycine aspartate: involvement of a tyrosine kinase-dependent, MAP kinase-independent mechanism. AB - Extracellular matrix facilitates anchorage-dependent cell survival via interaction of its arginine-glycine-aspartate (RGD) motif with integrins. In this report, we describe an unexpected, apoptosis-promoting the effect of immobilized RGD (iRGD) on tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis. Mesangial cells cultured on RGD-coated plates showed enhanced susceptibility to TNF-alpha-induced apoptosis. iRGD alone did not affect cell survival. In contrast, iRGD did not facilitate but inhibited apoptosis induced by H(2)O(2). Mitogen-activated protein (MAP) kinases and tyrosine kinases are important mediators for the RGD-integrin signaling. Pretreatment with MAP kinase kinase inhibitor PD098059, c-Jun N-terminal kinase (JNK)-c-Jun/AP-1 inhibitor curcumin or p38 MAP kinase inhibitor SB203580 did not attenuate the apoptosis-promoting effect of iRGD. Consistently, transfection with dominant-negative mutants of extracellular signal-regulated kinases, JNK or p38 MAP kinase did not inhibit the effect of iRGD. In contrast, protein tyrosine kinase inhibitors, genistein, and herbimycin A, abrogated the apoptosis-promoting effect of iRGD. Of note, TNF alpha-induced apoptosis on uncoated plates was not attenuated by tyrosine kinase inhibitors. These data provide the first evidence that iRGD accelerates certain apoptosis. We identified that the effect was mediated by the tyrosine kinase dependent, MAP kinase-independent mechanism. PMID- 11032721 TI - Regulation of the expression of the sea urchin mitochondrial D-loop binding protein during early development. AB - The Paracentrotus lividus mitochondrial D-loop binding protein (mtDBP) is a DNA binding protein which is involved in the regulation of sea urchin mtDNA transcription. Immunoblots of Heparin Sepharose-bound proteins at selected early developmental stages, as well as electrophoretic mobility shift assay, show that mtDBP is present in the egg at a concentration of about 1 x 10(6) molecules/egg. Its level increases after fertilization of about twofold, remaining substantially unchanged between 16-h blastula stage and early pluteus stage and declines thereafter. The content of mtDBP mRNA, determined by RNase protection experiments, increases about sevenfold at the 16-h blastula stage compared to the egg. A considerable decrease occurs at the 40-h pluteus stage, which precedes that of the protein. These results suggest that the expression of mtDBP is regulated at transcriptional level up to blastula stage, while other factors, in addition to the level of the RNA, may control the content of this protein in the following stages of embryogenesis. PMID- 11032722 TI - Gene responses in bean leaves induced by herbivory and by herbivore-induced volatiles. AB - Plant-plant interactions via herbivory-induced leaf volatiles could result in the induction of defense responses against aggressive biotic agents in plants. In this study, cDNA microarray technology showed comprehensive gene activation in lima bean leaves that were exposed to volatiles released from the neighboring leaves infested with spider mites. The infestation with spider mites and the herbivory-induced volatiles enhanced 97 and 227 gene spots on the microarray tip printed with 2032 lima bean cDNA, respectively. These genes are related to such broad functions as responses to pathogenesis, wounding, hormones, ethylene biosynthesis, flavonoid biosynthesis, (post) transcriptional modifications, translations, chaperones, secondary signaling messengers, membrane transports, protein/peptide degradations, and photosynthesis. We therefore conclude that herbivorous damage and herbivory-induced volatiles elicit comprehensive and drastic changes of metabolisms in leaves. PMID- 11032723 TI - Reconstitution of caspase-8 sensitizes JB6 cells to TRAIL. AB - TRAIL induces apoptosis in various tumor cells. We report here that caspase-8 is required in TRAIL-induced cell death. Western blot analyses and enzyme assays showed that exposing Jurkat cells to TRAIL resulted in activation of caspases-8 followed by caspase-3 and -9. Acetyl-IETD-fluoromethylketone, a caspase-8 inhibitor, potently suppressed TRAIL-induced cell death compared to acetyl-DEVD fluoromethylketone and acetyl-LEHD-fluoromethylketone, inhibitors of caspase-3 and caspase-9, respectively. JB6 cells, a caspase-8-deficient Jurkat variant, were completely resistant to TRAIL. However, reconstitution with a caspase-8, but not with caspase-2 or -3, sensitized JB6 cells to subsequent exposure to TRAIL. These results are indicative of the crucial function of caspase-8 in TRAIL induced apoptosis in Jurkat cells. PMID- 11032724 TI - Augmentation of signal transducer and activation of transcription (STAT)6 and STAT3 expression in stimulated B and T cells. AB - Signal transducer and activation of transcription (STAT)6 has a central role in the signal transduction of interleukin (IL)-4 and IL-13. It has recently been revealed that STAT3 is also involved. STAT6 and STAT3 are expressed ubiquitously; however, it remains unknown how STAT6 and STAT3 expression is regulated. In this study, we found that STAT6 expression was augmented at the transcription level in B and T cells stimulated with anti-IgM antibody and anti-CD40 antibody or PMA and ionomycin, respectively, and that STAT3 expression was similarly augmented in the stimulated B cells. The stimulated B and T cells showed enhancement of STAT6 activation and CD23 expression induced by IL-4 and IL-13. Augmentation of STAT6 and STAT3 would be a mechanism of the amplification of the IL-4 and IL-13 signals in stimulated B and T cells. PMID- 11032725 TI - Identification of ubiquitin as an immunophilin. AB - We have identified an 8.4 kDa minor immunophilin from calf thymus and Jurkat T cells as ubiquitin. It binds tacrolimus and sirolimus with K(d)'s of 0.8 and 0.08 nM, respectively. The binding of this protein to cyclosporin A is negligible. Binding of tacrolimus to two commercial sources of ubiquitin, a bovine product, and a recombinant human ubiquitin, was also demonstrated after HPLC purification of the purchased preparations. PMID- 11032726 TI - E-73, an acetoxyl analogue of cycloheximide, blocks the tumor necrosis factor induced NF-kappaB signaling pathway. AB - Proinflammatory cytokines such as tumor necrosis factor (TNF) and interleukin (IL)-1 activate the NF-kappaB signaling pathway which induces the expression of a variety of genes such as the encoding intercellular adhesion molecule (ICAM)-1. We have found that E-73, an acetoxyl analogue of cycloheximide, specifically blocks TNF-induced ICAM-1 expression even at concentrations unable to affect protein synthesis. By contrast, cycloheximide inhibited both TNF- and IL-1 induced ICAM-1 expression primarily due to the blockage of protein synthesis. The nuclear translocation of NF-kappaB as well as the IkappaB degradation induced by TNF, but not by IL-1, was significantly prevented by E-73. These observations suggest that E-73 blocks the TNF-induced NF-kappaB signaling pathway upstream of IkappaB degradation. PMID- 11032727 TI - gamma-tocopherol partially protects insulin-secreting cells against functional inhibition by nitric oxide. AB - Preceding the onset of type 1 diabetes mellitus, pancreatic islets are infiltrated by macrophages secreting interleukin-1beta (IL-1beta) which induces beta-cell apoptosis and exerts inhibitory actions on islet beta-cell insulin secretion. IL-1beta seems to act chiefly through induction of nitric oxide (NO) synthesis. Hence, IL-1beta and NO have been implicated as key effector molecules in type 1 diabetes mellitus. In this paper, the influence of endogenously produced and exogenously delivered NO on the regulation of cell proliferation, cell viability and discrete parts of the stimulus-secretion coupling in insulin secreting RINm5F cells was investigated. Because vitamin E may delay diabetes onset in animal models, we also investigated whether tocopherols may protect beta cells from the suppressive actions of IL-1 and NO in vitro. To this end, the impact of NO on insulin secretory responses to activation of phospholipase C (by carbamylcholine), protein kinase C (by phorbol ester), adenylyl cyclase (by forskolin), and Ca(2+) influx through voltage-activated Ca(2+) channels (by K(+) induced depolarization) was monitored in culture after treatment with IL-1beta or by co-incubation with the NO donor spermine-NONOate. It was found that cell proliferation, viability, insulin production and the stimulation of insulin release evoked by carbamylcholine and phorbol ester were impeded by IL-1beta or spermine-NONOate, whereas the hormone output by the other secretagogues was not altered by NO. Pretreatment with gamma-tocopherol (but not alpha-tocopherol) afforded a partial protection against the inhibitory effects of NO, whereas specifically inhibiting inducible NO synthase with N-nitro-L-arginine completely reversed the IL-1beta effects. In contrast, inhibiting guanylyl cyclase with ODQ (1H-[1,2, 4]oxadiazolo[4,3-alpha]-quinoxaline-1-one) or blocking low voltage activated Ca(2+) channels with NiCl(2) failed to influence the actions of NO. In conclusion, our data show that NO inhibits growth and insulin secretion in RINm5F cells, and that gamma-tocopherol may partially prevent this. The results suggest that phospholipase C or protein kinase C may be targeted by NO. In contrast, cGMP or low voltage-activated Ca(2+) channels appear not to mediate the toxicity of NO in these cells. These adverse effects of NO on the beta-cell, and the protection by gamma-tocopherol, may be of importance for the development of the impaired insulin secretion characterizing type 1 diabetes mellitus, and offer possibilities for intervention in this process. PMID- 11032728 TI - Neural BC1 RNA associates with pur alpha, a single-stranded DNA and RNA binding protein, which is involved in the transcription of the BC1 RNA gene. AB - BC1 RNA is preferentially expressed in neural cells by RNA polymerase III (Pol III) and forms ribonucleoprotein particles (RNP) in the somatodendritic domain of neurons. Our previous studies have suggested that, in the nucleus, BC1 RNA forms an RNP containing a nuclear protein(s) that participates in the transcription of the BC1 RNA gene. In this study, we have shown that newly synthesized BC1 RNA in purified brain nuclear extracts is immunoprecipitated by an antibody against Pur alpha. Pur alpha is a protein that binds single-stranded DNA and RNA and is known to regulate transcription of Pol II system. Although BC1 RNA is transcribed by Pol III, the BC1 RNA gene has two putative Pur alpha binding sites, which Pur alpha specifically recognizes. Point mutations within these sites reduced transcriptional activity in vitro. Furthermore, transcription was inhibited by depletion of Pur alpha from the nuclear extracts, either by the coexistence of its binding region of BC1 RNA or by the antibody that was able to precipitate the nuclear BC1 RNP. These observations suggest that BC1 RNA associates with Pur alpha which is involved in the transcription of the BC1 RNA gene. PMID- 11032729 TI - Tissue distribution of constitutive proteasomes, immunoproteasomes, and PA28 in rats. AB - We investigated the expression of standard proteasomes, immunoproteasomes, and their regulators, PA28, and PA700, in rat tissues. Immunoproteasomes (with subunits LMP2, LMP7, and MECL1) were abundant in the spleen but almost absent in the brain. In contrast, standard proteasomes (with X, Y, and Z) were highly expressed in the brain but not in the spleen. Both proteasome types were present in the lung and the liver. PA700 subunits (p112, S5a, and p45) were found in all tissues. PA28alpha, PA28beta, and PA28gamma were also expressed in all tissues, except for the brain which contained very little PA28beta. The results did not depend on rat sex or age. The cleavage specificity for peptide substrates differed greatly between brain and spleen proteasomes. Hybrid proteasomes, containing both PA28alphabeta and PA700, were not present in the brain but in all other tissues examined. PMID- 11032730 TI - FGF-20, a novel neurotrophic factor, preferentially expressed in the substantia nigra pars compacta of rat brain. AB - We have isolated cDNA encoding a novel FGF (212 amino acids) from rat brain. Because this is the 20th documented member of the FGF family, we tentatively term it FGF-20. Among FGF family members, FGF-20 is most similar to FGF-9 and FGF-16 (70 and 62% amino acid identity, respectively). Human FGF-20 gene was found in the human genomic sequence mapped to the 8p21.3-p22 region. Human FGF-20 is highly identical to rat FGF-20 (95% amino acid identity). FGF-20 mRNA was preferentially expressed in rat brain among the adult major tissues examined. The localization of FGF-20 mRNA in rat brain was also examined by in situ hybridization. FGF-20 mRNA was preferentially expressed in the substantia nigra pars compacta. To examine the biological activity of FGF-20, recombinant rat FGF 20 was produced by insect cells infected with recombinant baculovirus containing rat FGF-20 cDNA. Recombinant rat FGF-20 enhanced the survival of midbrain dopaminergic neurons. The present results indicate that FGF-20 is a novel neurotrophic factor preferentially expressed in the substantia nigra pars compacta of rat brain. PMID- 11032731 TI - The role of atypical and conventional PKC in dehydroepiandrosterone-induced glucose uptake and dexamethasone-induced insulin resistance. AB - We have reported that both dehydroepiandrosterone (DHEA) and dexamethasone (Dexa) directly activate PKC. In this study, we investigated the effects of these hormones on conventional PKC (cPKC) and atypical PKC (aPKC). DHEA and Dexa directly activated PKCbeta and PKCzeta to the same degree. In rat adipocytes, DHEA and Dexa activated endogenous immunoprecitable PKCzeta to 246 and 164%, respectively, from basal level (100%). In adipocytes, 5 min treatment with DHEA increased phosphatidylinositol 3-kinase (PI 3-kinase) activity in immunoprecipitate with anti-phosphotyrotyrosine antibody to 235%. Preincubation with wortmannin, myristoylated PKCzeta pseudosubstrate, but not with Go6976, abolished DHEA-induced 2-deoxyglucose (DOG) uptake. cPKC inhibitors prevented Dexa-induced insulin resistance. Moreover, DHEA and Dexa increased DOG uptake to 330 and 220%, respectively, in adipocytes overexpressed with wild-type PKCzeta, but not in those overexpressed with dominant negative. These results indicate that DHEA and Dexa activate both cPKC and aPKC, and Dexa-induced cPKC activation may lead to insulin resistance. In contrast, DHEA may mimic or enhance insulin action via PI 3-kinase and aPKC. PMID- 11032732 TI - Advanced glycation end products-induced gene expression of scavenger receptors in cultured human monocyte-derived macrophages. AB - We investigated the effects of advanced glycation end products (AGEs) on the expression of oxidized low-density lipoprotein (OxLDL) receptors in human monocyte-derived macrophages and THP-1 cells treated with PMA. Both RT-PCR procedure and Northern blot analysis revealed that AGEs induced not only the gene expression of two major OxLDL receptors, macrophage scavenger receptor (MSR) class A and CD36, but also MSR-B I and lectin-like oxidized low-density lipoprotein receptor 1. Also, as a result of gel shift assay, AGEs increased transcriptional activities of AP-1, NF-kappaB, and peroxisome proliferator activated receptor gamma. These findings indicate that AGEs-induced enhancement of these transcriptional activities might be involved in increased levels of mRNA for some of OxLDL receptors in THP-1-cells treated with PMA. The upregulated surface expression of these receptors on macrophage membranes was closely associated with increased uptake of modified LDL, and culminated in enhanced foam cell transformation. Thus, AGEs may be involved in the cause of variable levels of foam cell formation via the increased numbers of OxLDL receptors in accelerated atherosclerotic lesions of individuals with diabetes. PMID- 11032733 TI - Evaluation of the energy of red blood cell agglutination by monoclonal antibodies. AB - The knowledge of the energy involved in cell-cell interactions has significant implications in biological and medical sciences. Red blood cells (RBC) become mutually adhered when specific (agglutinins) or nonspecific macromolecules bind adjacent cells in an irreversible or reversible form. Flow chamber technique with digital image processing was successfully applied to determine the partial separation, by shear stress, of two RBC agglutinated face to face (doublet) by specific monoclonal antibodies (MAb) and also to evaluate the shear stress required to attain this separation. The work done in separating adhered cells is then easily calculated and serves to evaluate the antibody affinity. In this work, this technique was applied to evaluate the affinity of a monoclonal antibody specific to the blood group A antigen. The specific disaggregation energy gamma (i.e., the work done by the shear stress) required to disrupt a unit of adhered membrane areas between agglutinated cells was calculated. On play back of the image analysis, measurements were taken to determine the force applied to the doublet and the relative separation between both RBC. Values of gamma(d) (from 2.70 to 4.61 x 10(-9) N/cm) were found to be proportional to the density (D = 27 to 55 molecules/microm(2)) of MAb molecules bound on the RBC membrane. PMID- 11032734 TI - Formic acid dissolves aggregates of an N-terminal huntingtin fragment containing an expanded polyglutamine tract: applying to quantification of protein components of the aggregates. AB - Huntington's disease (HD) is caused by an expansion of the CAG repeat that encodes polyglutamine in huntingtin. Transient expression of an N-terminal huntingtin fragment containing an expanded polyglutamine tract induced formation of protein aggregates in cultured cells. The turnover of protein components in such aggregates has been difficult to study because of their insolubility in aqueous solutions. Here we describe a method of solubilizing the aggregates and quantifying their protein components. Insoluble pellets were collected from COS7 cells expressing an N-terminal huntingtin fragment containing an expanded polyglutamine tract and subjected to treatment with various detergent, acid, and alkaline reagents. Treatment with 100% formic acid at 37 degrees C for 30 min induced essentially complete dissociation of the aggregates to monomer. We used this solubilization technique to quantify huntingtin fusion protein in the aggregates formed in transient expression experiments. The frequency of aggregate formation increased when the proteasome inhibitor beta-lactone was added to culture media. However, the total amount of accumulated huntingtin fusion protein did not differ between cells cultured with or without beta-lactone. These results suggest that other protein components which are degraded by the proteasome, in addition to huntingtin, might be related to the dynamics of polyglutamine protein aggregates. PMID- 11032735 TI - The role of cysteine residues in tellurite resistance mediated by the TehAB determinant. AB - TehATehB is a tellurite (TeO(2-)(3)) resistance determinant found on the Escherichia coli chromosome. Normally silent, it specifies a minimal inhibitory concentration (MIC) of 2 microg K(2)TeO(3)/ml unless upregulated or present on a multicopy plasmid which results in an MIC of 128 microg/ml. Both TehA and TehB have three cysteine residues. Oligonucleotide site-directed mutagenesis was carried out to systematically replace all six cysteine residues by alaninies. The results showed that cysteine residues in both TehA and TehB play a role in tellurite resistance: A single cysteine change had no effect, however increasing combinations of two or three cysteine substitutions demonstrated strong phenotypic effects with minimal inhibitory concentrations ranging from 16-64 microg K(2)TeO(3)/ml. A cysteine-free mutant in which all six cysteine residues were replaced by alanines maintained a MIC of 16 microg/ml. Further investigations on the role of cysteines in resistance were studied using thiol reactive reagents on the soluble subunit TehB. These studies confirmed that TehB is a dimer and undergoes a conformational change with tellurite and S-adenosyl-l methionine binding. Studies using native and SDS denaturing PAGE under reducing and oxidizing conditions suggested that a cysteine in TehB is involved in binding tellurite. PMID- 11032736 TI - cDNA representational difference analysis of human neutrophils stimulated by GM CSF. AB - Neutrophils are the first cell type to migrate out of the vascular space and into the inflammatory site during an acute inflammation. However, in chronic inflammatory diseases, such as chronic obstructive pulmonary disease (COPD), a lack of clearance of neutrophils, imbalance between inflammatory mediators produced by neutrophils and their natural inhibitors make these cells a potential cause of tissue destruction in lung disease. Neutrophilic inflammation is generally characterised by high levels of local expression of activating cytokines (e.g., GM-CSF). Only a few studies have been published so far that have investigated the expression of genes preferentially expressed in activated neutrophils. The isolation of such genes, however, can lead to a better understanding of inflammatory disease and the identification of potential novel therapeutic targets or markers of the disease. We performed representational difference analysis of cDNA, a sensitive PCR-based subtractive enrichment procedure, and isolated 12 genes, 1 EST clone, and 3 sequences not represented in the public databases. Differential expression for 9 of these clones was confirmed by Northern hybridisation. Of the above nine transcripts three were chosen and shown to be up-regulated in neutrophils cocultured with stimulated primary human bronchial epithelial cells using a semiquantitative RT-PCR approach. Among the known genes identified were HM-74, CIS1, Cathepsin C, alpha-enolase, CD44, and the gene Translocation Three Four (TTF), most of them previously not known to be involved in GM-CSF induced neutrophil activation. Along with its tissue and cellular distribution we also derived the complete cDNA sequence and genomic structure of CIS1 using an in silico approach. In addition, we also report the initial characterisation of a novel gene, P1-89 that is primarily expressed in granulocytes and is up-regulated in activated cells. Our results identify several important genes associated with neutrophil activation and can lead to a better understanding of the molecular mechanisms of neutrophilic inflammations. PMID- 11032737 TI - Impaired angiogenic balance and suppression of tumorigenicity in HeLa cells chronically exposed to interferon-alpha. AB - We have previously reported that IFNalpha-chronic treatment for 41 days induced a partial phenotype reversion on HeLa cells along with a down-regulation of HPV18 mRNA levels. However, tumorigenicity of these cells in nude mice was unchanged. Interestingly, after 1 year of IFNalpha-chronic exposition, HeLa cells failed to induce s.c. tumors when injected into nude mice. In such experimental conditions both HPV18 DNA integration pattern and viral DNA copy number present in HeLa cells remained intact in the nontumorigenic phenotype cells. As result of the treatment with IFNalpha, HeLa cells rendered more resistant to lysis mediated by activated natural killer cells in vitro. Furthermore, IFNalpha-chronic treatment was able to induce VEGF and decrease bFGF mRNA expression, suggesting a potential effect on the angiogenic behavior of these tumoral cells. Thus, long-term treatment of HeLa cells with IFNalpha can accomplish a reversion of the malignant phenotype by a sequential multistep mechanism, in which the antiangiogenic effect of IFNalpha could be one of the contributing events. PMID- 11032738 TI - Glibenclamide acts as an inhibitor of acyl-CoA:cholesterol acyltransferase enzyme. AB - Sulfonylureas are used in the treatment of non-insulin-dependent diabetes mellitus. Little is known, however, about their effects on cholesterol metabolism. We tested in the present study the effects of glibenclamide (GB) on cholesterol esterification (CE) in macrophage-derived cells. GB inhibited intracellular accumulation of CE induced by acetylated LDL or oxidized LDL in J774 cells, but no such effect on total cholesterol, suggesting that the target of GB was acyl-CoA:cholesterol acyltransferase (ACAT). In the cell-free reconstitution ACAT assay, GB inhibited the ACAT activity with an IC(50) value of 20 microM. Furthermore, GB effectively inhibited the ACAT activity of PMA stimulated THP-1 cells to the undifferentiated level of THP-1. In the whole-cell ACAT assay using CHO cells overexpressed with ACAT-1 or ACAT-2, GB inhibited the activity of both isozymes with similar potency. Our in vitro data suggest that sulfonylurea could be a potential seed for a new generation of ACAT inhibitors. PMID- 11032739 TI - Differential expression of lipoprotein lipase gene in tissues of the rat model with visceral obesity and postprandial hyperlipidemia. AB - Postprandial hyperlipidemia is frequently accompanied with intra-abdominal visceral accumulation in human subjects. We have found that the decreased lipoprotein lipase (LPL) mass and activity is negatively associated with the amount of visceral fat accumulation. Here, we studied the postprandial hyperlipidemia using the OLETF rat, a model with visceral obesity, in order to clarify the molecular mechanism causing postprandial hyperlipidemia accompanied with visceral obesity. At the same age of 32 weeks, the OLETF rats showed obviously higher plasma leptin, total cholesterol, triglyceride, and HDL cholesterol levels than the control LETO rats, although the plasma glucose level was not significantly different. Fat-loading test revealed the delayed metabolism of exogenous fat in the OLETF rats compared to the LETO rats, similar to human subjects with visceral obesity. In the obese rats, plasma levels of LPL mass and activities were 60 and 49% of control rats. The expression of LPL gene was decreased in subcutaneous adipose tissues and skeletal muscle of OLETF rats to 40 and 52% compared to those of LETO rats. In OLETF rats, plasma tumor necrosis factor-alpha (TNF-alpha) and insulin levels were increased to 2.0- and 2.3-folds compared to those in control rats. Furthermore, plasma insulin and TNF-alpha levels in OLETF rats were negatively correlated with the expression levels of LPL gene in subcutaneous fat and muscle. These results indicate that decreased LPL mass and activity in the animal model with visceral obesity is possibly caused by decreased expression of LPL gene in tissues mediated by the increased levels of insulin and TNF-alpha. The different expression of LPL gene in tissues associated with the increased levels of insulin and TNF-alpha possibly elucidate the underlying mechanisms involving the postprandial hyperlipidemia observed in visceral obesity. PMID- 11032740 TI - Immunochemical demonstration of a novel beta-subunit isoform of X, K-ATPase in human skeletal muscle. AB - Recently we have identified mRNA encoding a hitherto unknown mammalian X,K-ATPase beta-subunit expressed predominantly in muscle tissue (Pestov, N. B. et al. (1999) FEBS Lett. 456, 243-248). Here we demonstrate the existence of the predicted protein, designated as beta(m) (beta(muscle)), in human adult skeletal muscle membranes using immunoblotting with beta(m)-specific antibodies generated against recombinant polypeptide formed by extramembrane beta(m) domains. The electrophoretic mobility of beta(m) was shown to be abnormally low due to the presence of Glu-rich sequences. In contrast to mature forms of other known X,K ATPase beta-subunits, carbohydrate moiety of beta(m) is sensitive to endoglycosidase H and appears to be composed of short high-mannose or hybrid N glycans. This finding argues in favor of an intracellular location of beta(m) in human skeletal muscle. PMID- 11032741 TI - Protein translocation within chloroplast is similar in Euglena and higher plants. AB - It is currently thought that chloroplasts of higher plants were derived from endosymbiont oxygenic photosynthetic bacteria (primary endosymbiosis), while Euglena, a photosynthetic protista, gained chloroplasts by secondary endosymbiosis (i.e., incorporation of a photosynthetic eukaryote into heterotrophic eukaryotic host). To examine if the protein transport inside chloroplasts is similar between these organisms, we carried out heterologous protein import experiments with Euglena precursor proteins and spinach chloroplasts. The precursor of a 30-kDa subunit of the oxygen-evolving complex (OEC30) from the thylakoid lumen of Euglena chloroplasts contained the N-terminal signal, stroma targeting, and thylakoid transfer domains. Truncated preOEC30s lacking the N-terminal domain were post-translationally imported into spinach chloroplasts, transported into the thylakoid lumen, and processed to a mature protein. These results showed that protein translocations within chloroplasts in Euglena and higher plants are similar and supported the hypothesis that Euglena chloroplasts are derived from the ancestral Chlorophyta. PMID- 11032742 TI - Thioredoxin, a singlet oxygen quencher and hydroxyl radical scavenger: redox independent functions. AB - Thioredoxin is a ubiquitous small protein known to protect cells and tissues against oxidative stress. However, its exact antioxidant nature has not been elucidated. In this report, we present evidence that human thioredoxin is a powerful singlet oxygen quencher and hydroxyl radical scavenger. Human thioredoxin at 3 microM caused 50% inhibition of TEMP-(1)O(2) (TEMPO) adduct formation in a photolysis EPR study. In contrast, Escherichia coli thioredoxin caused 50% inhibition of TEMPO formation at 80 microM. Both E. coli thioredoxin and human thioredoxin inhibited (*)OH dependent DMPO-OH formation as demonstrated by EPR spectrometry. The quenching of (1)O(2) or scavenging of (*)OH was not dependent upon the redox state of thioredoxin. Using a human thioredoxin in which the structural cysteines were mutated to alanine, Trx-C3A, we show that structural cysteines that do not take part in the catalytic functions of the protein are also important for its reactive oxygen scavenging properties. In addition, using a quadruple mutant Trx-C4A, where one of the catalytic cysteines, C35 was mutated to alanine in addition to the mutated structural cysteines, we demonstrated that catalytic cysteines are also required for the scavenging action of thioredoxin. Identification of thioredoxin as a (1)O(2) quencher and (*)OH scavenger may be of significant importance in explaining various redox-related antioxidant functions of thioredoxin. PMID- 11032743 TI - Kinetics of expression of connective tissue growth factor gene during liver regeneration after partial hepatectomy and D-galactosamine-induced liver injury in rats. AB - Connective tissue growth factor (CTGF) is up-regulated by TGF-beta1 during wound healing. The present study examined the expression of CTGF during regeneration after 70% partial hepatectomy (PH) or d-galactosamine (GalN)-injured liver in rats. CTGF, TGF-beta1, and type I collagen mRNAs were semiquantified by a ribonuclease protection assay. After PH, TGF-beta1 and type I collagen were increased at 2-6 h and at 12-48 h. CTGF increased at 6 h and returned to the control level thereafter. The ribonuclease protection assay of cultured hepatic stellate cells (HSC) and in situ hybridization suggest that the cells express CTGF along sinusoid might be HSCs. After GalN administration, CTGF increased at 2 96 h with a shoulder peak at 6-12 h followed by a main peak at 24 h. TGF-beta1 and type I collagen were up-regulated with kinetics similar to those of CTGF. The different kinetics between PH and GalN regenerations indicate that regulation of CTGF in the two processes is different. Higher TGF-beta1 expression after inflammatory/necrotic process in the GalN regeneration may caused the prolonged CTGF expression. PMID- 11032744 TI - Phosphatidylinositol 3-kinase signaling to Akt mediates survival in isolated canine islets of Langerhans. AB - The isolation of islet cells from the pancreas by enzymatic digestion causes many of these cells to undergo apoptosis. The aim of this work was to investigate the role of phosphatidylinositol 3-kinase (PI3-K)/Akt signaling in mediating the survival of isolated islets. Insulin-like growth factor-1 (IGF-I) was examined as a potential culture media supplement that could rescue isolated islets from their apoptotic fate. Western blot analysis demonstrated that Akt phosphorylation peaks 20 h after routine islet isolation. PI3-K inhibition with wortmannin abolished both basal and IGF-I-mediated Akt phosphorylation. IGF-I did not increase survival of isolated islets under normal conditions but it did have a protective effect against cytokine (TNF-alpha, IL-1beta, INF-gamma)-mediated cell death. The protective effect of IGF-I against cytokine-stimulated apoptosis was blocked by wortmannin. In addition, inhibition of basal levels of PI3-K activity caused a 31% decrease in islet survival, as shown by MTT assay. These results demonstrate that the PI3-K/Akt pathway mediates survival of isolated islets of Langerhans. PMID- 11032745 TI - Cellular uptake and in situ binding of a peptide agonist for calmodulin. AB - We have used the method of inverted hydropathy to develop peptides that interact with EF hands of calmodulin (CaM). Previously we have shown these peptides specifically interact with their desired target in a productive manner, in that they activated CaM in the absence of Ca(2+). Therefore, we sought to determine whether these peptides would enter cells, remain intact, and interact with CaM in the interior of the cell. Using several techniques we have demonstrated cellular uptake, stability, and an intracellular interaction with CaM with fluorescein labeled and radiolabeled peptides in Jurkat T cells. The results suggest that these peptides may be useful in the study and the manipulation of Ca(2+)-mediated pathways in cells. PMID- 11032746 TI - Protective effect of melatonin against homocysteine-induced vasoconstriction of human umbilical artery. AB - Hyperhomocysteinemia is a major and independent risk factor for vascular disease. Oxidative stress is a possible mechanism for homocysteine (Hcy)-induced endothelial dysfunction. Herein, we evaluated the antioxidant property of melatonin (MLT) in relation to the vasoconstrictive effect of Hcy on the human umbilical artery. In an initial experiment in a cell-free system, a micromolar concentration of iron was found to catalyze oxygen-dependent oxidation of Hcy. MLT (10 or 100 microM) did not affect oxygen-dependent oxidation of Hcy. Next, smooth muscle contraction induced by prostaglandin F(2alpha) (10 microM) was measured in arterial strips. Hcy (10 to 500 microM) increased this vascular tension in a concentration-dependent manner (P < 0.0001). Addition of Fe(2+) (10 microM) significantly potentiated the Hcy effect. Removal of endothelium (P < 0.05), pretreatment with a nitric oxide (NO) synthesis inhibitor (l-N(G) monomethylarginine, 200 microM, P < 0.001), or pretreatment with a hydroxyl radical ((*)OH) scavenger (mannitol, 10 mM, P < 0.001) significantly attenuated contraction potentiated by Hcy plus Fe(2+). At a much lower concentration than mannitol, MLT (1 to 100 microM) significantly reduced the contractile effect of Hcy and Fe(2+) in a concentration-dependent manner. Hcy plus Fe(2+) significantly impaired calcium ionophore A 23187-induced relaxation (P < 0.0001), while MLT restored this relaxation in a concentration-dependent manner. These findings suggest that Hcy potentiates vascular tension in human umbilical artery, possibly by suppressing bioavailable NO. MLT protects against the vasoconstrictive effect of Hcy, most likely by scavenging (*)OH arising from Hcy autooxidation. PMID- 11032747 TI - Differential gene expression in apoptosis: identification of ribosomal protein S29 as an apoptotic inducer. AB - To identify genes that are specifically involved in apoptosis, poly(A)(+) RNAs were isolated from untreated control rat thymocytes and from adriamycin-induced apoptotic thymocytes. Directionally cloned cDNA libraries were then constructed in UNIZAP-XR vectors followed by biotin-based subtractive hybridization. Three clones were confirmed to be differentially expressed by dot blotting. Sequence analysis revealed homology to two genes previously identified, whereas one clone was novel and did not have homology to any known sequence. One clone was identical to the ribosomal protein S29, and the other was homologous to L8 ribosomal protein. Northern blot analysis revealed a marked increase in the expression of mRNA encoding ribosomal protein S29 in the apoptotic thymocytes compared to the controls. Transfection studies revealed that enhanced S29 expression resulted in increased apoptosis in rat thymocytes and HeLa cells as assessed by various morphological and biochemical characteristics, including cell shrinkage, chromatin condensation, membrane blebbing, formation of apoptotic bodies, TUNEL, FACS, and internucleosomal DNA fragmentation. This was accompanied by upregulation of p53, Caspase 3, and bax, whereas bcl-2 was downregulated as revealed by Western blotting. The current findings provide the first hint of a role for ribosomal protein S29 in the apoptotic process. PMID- 11032748 TI - Escape from apoptosis after prolonged serum deprivation is associated with the regulation of the mitochondrial death pathway by Bcl-x(l). AB - Bcl-x(l) and Bax play important roles in the regulation of apoptosis. This study investigated the involvement of the mitochondrial death pathway and the role of Bcl-x(l) and Bax in the escape from apoptosis after prolonged serum deprivation in Madin-Darby canine kidney (MDCK) cells. Low level apoptosis and basal activity of the mitochondrial death pathway were detectable in normal cell growth. In serum deprivation, mitosis was partially suppressed, and the mitochondrial activity was stimulated. The level of apoptosis continuously rose over 48 h. This rise was concomitant with the increasing presence of cytochrome c in cytosol. However, both apoptosis and cytosolic cytochrome c fell dramatically at 72 h. Elevation of whole cell Bcl-x(l) and redistribution of Bcl-x(l) protein from cytosol to the membrane at 48 h and 72 h was observed. Redistribution of Bax protein from the membrane to cytosol occurred at 24 h, and remained steady to 72 h. Bax/Bcl-x(l) coimmunoprecipitation by anti-Bax antibody showed reduced Bax/Bcl x(l) interaction at the membrane at 72 h, but not at 24 or 48 h. These results suggest that apoptosis upon serum withdrawal results from the leakage of cytochrome c to cytosol. Amelioration of the leakage of cytochrome c and apoptosis requires not only the increase of Bcl-x(l)/Bax ratio, but also the release of Bcl-x(l) from Bax at the membrane. PMID- 11032749 TI - Identification of a novel fibroblast growth factor, FGF-23, preferentially expressed in the ventrolateral thalamic nucleus of the brain. AB - We isolated mouse cDNA encoding a novel FGF (251 amino acids). As this is the 23rd documented FGF, we termed it FGF-23. FGF-23 has a hydrophobic amino terminus ( approximately 24 amino acids), which is a typical signal sequence. As expected, recombinant mouse FGF-23 was efficiently secreted by High Five insect cell infected recombinant baculovirus containing the cDNA, indicating that FGF-23 is a secreted protein. We also isolated human cDNA encoding FGF-23 (251 amino acids), which is highly identical ( approximately 72% amino acid identity) to mouse FGF 23. Of human FGF family members, FGF-23 is most similar to FGF-21 and FGF-19 ( approximately 24% and approximately 22% amino acid identities, respectively). Human FGF-23 gene was localized on the chromosome 12p13 and found to be tandem linked (within 5.5 kb) to human FGF-6 gene. The expression of FGF-23 mRNA in mouse adult tissues was examined by real-time quantitative polymerase chain reaction. FGF-23 mRNA was mainly expressed in the brain and thymus at low levels. The localization of FGF-23 mRNA in the brain was examined by in situ hybridization. FGF-23 mRNA in the brain was found to be preferentially expressed in the ventrolateral thalamic nucleus. Therefore, FGF-23 is expected a unique FGF that plays roles in the function of the ventrolateral thalamic nucleus. PMID- 11032750 TI - Cytotoxic activity of recombinant bFGF-rViscumin fusion proteins. AB - A fusion protein (bFGF-rMLA), containing the mitogen basic fibroblast growth factor (bFGF) and the cytotoxic component of rViscumin (recombinant mistletoe lectin), the enzymatic A-chain (rMLA), was expressed in Escherichia coli, purified, and functionally characterized. bFGF-rMLA is cytotoxic for mouse B16 melanoma cells expressing the FGF receptor with an IC(50) value of approximately 1 nM. rMLA shows no significant effect on the viability of the B16 cells up to a concentration of 141 nM. Additionally, bFGF-rMLA was associated with the rViscumin B-chain (rMLB) in an in vitro folding procedure. The IC(50) value of bFGF-rMLA/rMLB to B16 cells in the presence of lactose-to block rMLB lectin activity-was 134 pM. Thus, it was possible to enhance the efficacy of a rViscumin A-chain mitotoxin through addition of rMLB. We conclude that rViscumin fusion proteins may be generally applicable for the receptor-specific inactivation of target cells and point out their potential in drug development. PMID- 11032751 TI - Epicatechin and catechin are O-methylated and glucuronidated in the small intestine. AB - There is considerable interest in the bioavailability of polyphenols and their bioactivity in vivo. We have studied the absorption and metabolism of catechin and epicatechin in the small intestine and the comparative transfer across the jejunum and ileum. Perfusion of isolated jejunum with the flavanols resulted in glucuronidation ( approximately 45%), O-methylation: 3'-O-Methyl- and 4'-O-methyl ( approximately 30%), and O-methyl-glucuronidation ( approximately 20% of total flavanols identified) during transfer across the enterocytes to the serosal side. This demonstrates the activity of catechol-O-methyl transferases in the metabolism of flavanols and suggests that these metabolites and conjugates are likely to enter the portal vein. In contrast, in the case of the ileum, the majority of the flavanols appeared on the serosal side unmetabolised and the total percentage of flavanols transferred was higher than that in the jejunum ( approximately fivefold). PMID- 11032752 TI - The serine/threonine kinase HIPK2 interacts with TRADD, but not with CD95 or TNF R1 in 293T cells. AB - HIPK2 has been described as a homoedomain-interacting protein kinase with a nuclear localization. Here we describe that HIPK2 can also associate with TRADD, a protein that interacts with tumor necrosis factor receptor type 1 (TNF-R1). Under the conditions where HIPK2/TRADD association was found, no direct interaction of HIPK2 with CD95, TNF-R1, FADD or caspase-8 could be detected. Therefore, HIPK2 may play a role in TNF-R1 mediated signaling. PMID- 11032753 TI - The CUG-binding protein binds specifically to UG dinucleotide repeats in a yeast three-hybrid system. AB - The CUG-binding protein (CUG-BP) has been reported to be involved in the pathogenesis of myotonic dystrophy (DM) through binding to a CUG trinucleotide repeat located in the 3' untranslated region (3'UTR) of the DM protein kinase (DMPK) gene. We found that CUG-BP associates with long CUG trinucleotide repeats ((CUG)(11)(CUG)(12)), but not with short repeats ((CUG)(12)) in a yeast three hybrid system. On the other hand, CUG-BP+LYLQ, an alternatively spliced isoform of CUG-BP, does not associate with CUG trinucleotide repeats regardless of the repeat length. In addition to these findings, we found that CUG-BP and CUG BP+LYLQ strongly and specifically associate with UG dinucleotide repeats. Deletion analyse of CUG-BP revealed that the absence of the first or third RNA binding domain (RBD I and RBD III, respectively) does not affect the interaction between CUG-BP and UG dinucleotide repeats. Loss of the second RNA-binding domain (RBD II) decreases the affinity of CUG-BP for UG dinucleotide repeats by about 40%. Unexpectedly, deletion of the linker domain most severely reduces the interaction, although this region does not contain a known RNA-binding motif. Our results suggest the possibility that both CUG-BP and CUG-BP+LYLQ associate with UG repeat-containing mRNAs and regulate such metabolic properties as mRNA localization, stability, and translation, and provide new insights into the pathogenesis of DM. PMID- 11032754 TI - Cellular responses to sodium butyrate exhibit the dominance of one parental phenotype in somatic cell hybrids. AB - The glycoprotein hormone alpha-subunit (GPHalpha) gene is inducible by sodium butyrate (NaBtr) in nontrophoblastic tumor cell lines such as HeLa (cervical carcinoma) but not in trophoblastic tumor cell lines such as JEG-3 (choriocarcinoma). The studies summarized in this report examined the ability of NaBtr to induce GPHalpha expression in somatic cell hybrids between HeLa SR3(hyg) and JEG-3(neo). The hybrid cells, pooled clones resistant to both hygromycin B and G418 sulfate, have been named JELA and were indistinguishable from the SR3 parent with regard to induction of the GPHalpha gene. The effects of NaBtr on cell proliferation were also similar in HeLa and JELA but different from those in JEG-3. The GPHalpha gene could be induced by NaBtr in the JEG-3 parent only when they were simultaneously treated with cycloheximide (CHX). The ability of NaBtr to induce GPHalpha in CHX-treated JEG-3 cells occurred concomitantly with a change in the electrophoretic mobility of enhancer binding proteins as determined in gel shift assays. The DNA-protein complexes generated between a trophoblast specific element (TSE) and nuclear proteins in HeLa SR3 and JELA migrated significantly more slowly than the complex generated by JEG-3 nuclear proteins. However, when nuclear extracts were prepared from CHX-treated JEG-3 cells, the complex generated with the TSE oligonucleotide migrated more slowly than the complex from untreated JEG-3 cells and coincident with the complexes produced with nuclear extracts from HeLa SR3 and JELA cells. Together, these data demonstrate that inducibility of the GPHalpha gene by NaBtr in JELA cell hybrids resembles that of the HeLa SR3 parent and that its inducibility in the JEG-3 parent parallels the status of an enhancer binding protein (TSEB) as judged from changes in electrophoretic mobility. The results are consistent with a model in which the status of TSEB has a profound influence on the gene's response to NaBtr. PMID- 11032755 TI - Normal breast epithelial cells induce apoptosis of MCF-7 breast cancer cells through a p53-mediated pathway. AB - Cancer development depends not only on the nature of the tumor cells themselves but also on the regulatory effects of various normal cells. The present study was performed to better understand the mechanism by which normal breast epithelial cells (NBEC) can control the growth of MCF-7 breast cancer cells. When MCF-7 cells were treated with NBEC conditioned medium, cell growth was inhibited in a concentration-dependent manner. This inhibition was due to an induction of apoptosis without any change in cell cycle progression. The induction of apoptosis was correlated with increased levels of p53, p21(waf1) and decreased levels of bcl-2. Transient transfections of MCF-7 cells with two p53 cDNA constructs demonstrated the induction of apoptosis was mediated by endogenous p53. Taken together, our results indicate that NBEC inhibit the growth of MCF-7 breast cancer cells by inducing apoptosis in them via endogenous p53. PMID- 11032756 TI - Expression and purification of recombinant human SPARC produced by baculovirus. AB - SPARC (secreted protein acidic and rich in cysteine/osteonectin/BM-40), a matrix associated protein, disrupts cell adhesion and inhibits the proliferation of many cultured cells. We report the expression of recombinant human protein (rhSPARC) in a baculovirus expression system. This procedure routinely yields approximately 1 mg of purified protein per 500 ml of culture supernate. rhSPARC produced by insect cells migrates at the appropriate molecular weight under reducing and nonreducing conditions. The rhSPARC purified from insect cell media appeared structurally similar to SPARC purified from mammalian tissue culture by the criterion of circular dichroism. In addition, a series of anti-SPARC and anti SPARC peptide antibodies recognized insect cell rhSPARC. We also show that rhSPARC produced in this system is glycosylated and is biologically active, as assessed by inhibition of endothelial cell proliferation and induction of collagen I mRNA in mesangial cells. Significant amounts of rhSPARC can now be generated in the absence of contaminating mammalian proteins for structure/function assays of SPARC activities. PMID- 11032757 TI - Downregulation of cyclin D1 alters cdk 4- and cdk 2-specific phosphorylation of retinoblastoma protein. AB - Progression of cells through the G1 phase of the cell cycle requires the assembly and activation of specific cyclin:cyclin-dependent kinase (cdk) complexes in a tightly regulated, sequential fashion. To more clearly define the temporal events leading to the G1/S transition, sequential changes in the expression of cyclin E and cdks 2, 4, and 6, as well as the phosphorylation of the retinoblastoma protein (pRb), were assayed in RA28 cells, a variant of human colon cancer RKO cells which were modified by transfection of an ecdysone-inducible antisense (AS) CD1 expression system. Induction of cyclin D1 antisense mRNA by the ecdysteroid, ponasterone A, resulted in a 55% decrease in cyclin D1 mRNA and a 58% decrease in CD1 protein levels. There was a 2.4-fold decrease in the ratio of hyperphosphorylated pRb (ppRb) to hypophosphorylated pRb, as well as a 60-75% decrease in cdk 2- and cdk 4-specific phosphorylated pRb proteins. Of interest, cyclin E-dependent phosphorylation (cdk2) decreased 2.5-fold at 3 h despite only a 30% decrease in cyclin E protein level. Levels of cdk 2, cdk 4, and cdk 6 decreased 40-70%, while levels of cyclin A and B were unaffected by induction of CD1 antisense. Induction of a CD1 antisense gene in a human colon cancer cell line resulted in rapid, concomitant changes in CD1 mRNA and protein, cyclin E, cdk2, cdk4, and cdk6, as well as the ratio of ppRb to pRb. In this system, growth regulatory events are tightly regulated and the perturbed expression of a single protein, CD1, rapidly alters expression of multiple regulatory proteins involved in the G1/S transition phase of cell cycle progression. PMID- 11032758 TI - Novel inhibitors of advanced glycation endproducts (part II). AB - Enhanced formation and accumulation of advanced glycation endproducts (AGEs), have been implicated as a major pathogenesis process leading to diabetic complications, normal aging, atherosclerosis, and Alzheimer's Disease. Several potential drug candidates as AGE inhibitors have been reported recently. The aim of this study was to develop classes of novel inhibitors of glycation, AGE formation, and AGE-crosslinking and to investigate their effects through in vitro chemical and immunochemical assays. A total of 92 compounds were designed and synthesized. The first 63 compounds were reported before. Nearly half of the 29 novel inhibitors reported here are benzoic acid derivatives and related molecules, and found to be potent inhibitors of multistage glycation, AGE formation, and AGE-protein crosslinking. All 29 compounds show some degrees of inhibitory activities as detected by the four assay methods, 9 compounds demonstrated high percent inhibition (PI) in all tests, 30 to 40 times stronger than aminoguanidine. PMID- 11032759 TI - Repression of transcription by HoxC11 upon phorbol ester stimulation. AB - Hox genes encode transcription factors with a conserved DNA-binding domain and exhibit similar DNA-binding preferences. The in vivo specificity required for their biological function is brought about by combinatorial interactions with other factors. Such interactions also modulate their activation state. Here we show that HoxC11 can either activate or repress transcription in a signal specific manner. We report the isolation of HoxC11 in a yeast one-hybrid screen for factors binding to a phorbol-ester, 12-O-tetradecanoylphorbol-13-acetate (TPA) response element (VLTRE), which is also a target for TPA-induced binding of Rel factors in gel-shift experiments. Although we detect no binding of in vitro translated HoxC11 to the TPA response element in EMSA, overexpression of HoxC11 in the HepG2 cell line leads to a complete block of TPA-induced transcription from a VLTRE-luciferase reporter. There is, however, no repression of the basal levels. The repression is furthermore not dependent on homeo-domain DNA binding. Our data suggest an interaction of HoxC11 with the basal-transcription machinery. We propose that HoxC11 is capable of mediating transcriptional activation or repression in a signal-specific manner and that its activation of the DNA target sequence in yeast might reflect in vivo recruitment to the promoter complex. PMID- 11032760 TI - Expression of intracellular calcium channels and pumps after partial hepatectomy in rat. AB - Ca(2+) signals regulate many cellular functions, including proliferation. They are governed by the inositol 1,4,5-trisphosphate receptor (IP(3)R), the only intracellular hepatic Ca(2+) channel and by the endoplasmic reticulum Ca(2+) pumps, SERCA. To characterise their role in regeneration, expression of their isoforms was studied after 2/3 hepatectomy by real-time quantitative PCR, Western blot and binding studies. We found an early increase in the expression of the IP(3)R isoform 1 which contrasted with the decrease of the expression of the IP(3)R isoforms 2 and 3 and of SERCA3. This results in a transient switch between IP(3)R isoforms 1 and 2, IP(3)R isoform 1 becoming predominant before the first round of mitosis. Binding studies detected a 30% diminution of the IP(3)R population at 24 h. In conclusion, the Ca(2+) signalling machinery is regulated, after hepatectomy, by changes in expression of the IP(3)R and SERCA isoforms to adapt Ca(2+) signals to the regenerative state. PMID- 11032761 TI - Advanced glycation end products induce blood-retinal barrier dysfunction in normoglycemic rats. AB - Advanced glycation end products (AGEs) have been implicated in the progressive vascular dysfunction which occurs during diabetic retinopathy. In the current study we have examined the role of these adducts in blood-retinal barrier (BRB) breakdown and investigated expression of the vasopermeabilizing agent vascular endothelial growth factor (VEGF) in the retina. When normoglycemic rats were injected with AGE-modified albumin daily for up to 10 days there was widespread leakage of FITC-dextran and serum albumin from the retinal vasculature when compared to control animals treated with nonmodified albumin. Ultrastructural examination of the vasculature revealed areas of attenuation of the retinal vascular endothelium and increased vesicular organelles only in the AGE-exposed rats. Quantitative RT-PCR and in situ hybridization demonstrated a significant increase in retinal VEGF mRNA expression (P < 0.05). These results suggest that AGEs can initiate BRB dysfunction in nondiabetic rats and a concomitant increase in retinal VEGF expression. These findings may have implications for the role of AGEs in the pathogenesis of diabetic retinopathy. PMID- 11032762 TI - p53 codon 72 polymorphism as a risk factor in the development of breast cancer. AB - The p53 gene is polymorphic at amino acid 72 of the protein that it encodes. It has been reported that patients with the arginine form have a higher risk of developing other forms of cancer than those with the proline form. The purpose of this study was to examine whether p53 Arg at the polymorphic position 72 could represent a risk factor for women with breast lesions. The study population included 56 biopsies from patients with breast lesions. Also, 61 normal blood samples were used as controls. There was a difference in the distribution of p53 genotypes between breast cancer lesions and the normal samples. The allele frequency of p53 Arg/Arg was much higher than that of the normal samples (61% versus 20%). Based on the findings of this study, it is suggested that p53 Arg homozygosity could represent a risk factor for the tumorigenesis of the breast. PMID- 11032763 TI - Effects of arsenic, cadmium, chromium, and lead on gene expression regulated by a battery of 13 different promoters in recombinant HepG2 cells. AB - Toxic metals occur naturally at low concentrations throughout the environment, but are found in higher concentrations at many of the hazardous waste sites on the EPA Superfund list. As part of the Agency for Toxic Substances and Disease Registry (ATSDR) mandate to evaluate the toxicity of metals and mixtures, we chose four of the high-priority metal pollutants from ATSDR's HAZDAT list, including arsenic, cadmium, chromium, and lead, to test in a commercially developed assay system, CAT-Tox(L) (Xenometrix). This assay employs a battery of recombinant HepG2 cell lines to test the transcriptional activation capacity of xenobiotics in any of 13 different signal transduction pathways. Our specific aims were to identify metal-responsive promoters and determine whether the pattern of gene expression changed with a mixture of metals. Humic acid was used in all assays as a carrier to help solubilize the metals and, in all cases, the cells were exposed to the humic acid-metal mixture for 48 h. Humic acid alone, at 50-100 microM, showed moderate activation of the XRE promoter, but little other notable activity. As(V), at doses of 50-250 microM, produced a complex profile of activity showing significant dose-dependent induction of the hMTIIA, GST Ya, HSP70, FOS, XRE, NFkappaBRE, GADD153, p53RE, and CRE promoters. Pb(II) showed dose-related induction of the GST Ya, XRE, hMTIIA, GRP78, and CYP IA1 promoters at doses in the range of 12-100 microM. Cd(II), at 1.25-15 microM, yielded significant dose-dependent induction of hMTIIA, XRE, CYP IA1, GST Ya, HSP70, NFkappaBRE, and FOS. Whereas Cr(III) yielded small, though significant inductions of the CRE, FOS, GADD153, and XRE promoters only at the highest dose (750 microM), Cr(VI) produced significant dose-related inductions of the p53RE, FOS, NFkappaBRE, XRE, GADD45, HSP70, and CRE promoters at much lower doses, in the range of 5-10 microM. Assays testing serial dilutions of a mixture comprising 7.5 microM Cd(II), 750 microM Cr(III), and 100 microM Pb(II) (the combination of metals most frequently found at National Priority List sites) showed significant dose-dependent induction of the hMTIIA promoter, but failed to show dose-related induction of any other promoter and showed no evidence of synergistic activation of gene expression by the metals in this mixture. Our results thus show metal activation of gene expression through several previously unreported signal transduction pathways, including As(V) induction of GST Ya, FOS, XRE, NFkBRE, GADD153, p53RE, and CRE; Pb(II) induction of GST Ya, XRE, Cyp IA1, and GADD153; Cd(II) induction of NFkBRE, Cyp IA1, XRE, and GST Ya; and Cr(VI) induction of p53RE, XRE, GADD45, HSP70, and CRE promoters, and thus suggest new insights into the biochemical mechanisms of toxicity and carcinogenicity of metals. It is also an important finding that no evidence of synergistic activity was detected with the mixture of Cd(II), Cr(III), and Pb(II) tested in these assays. PMID- 11032764 TI - Effects of 17beta-estradiol and testosterone on hepatic mRNA/protein levels and catalytic activities of CYP2M1, CYP2K1, and CYP3A27 in rainbow trout (Oncorhynchus mykiss). AB - There is growing concern that exposure to chemicals in the environment can disrupt the endocrine systems of wildlife and humans, causing reproductive problems or other adverse effects. The expression of many cytochrome P450s (CYPs) is under hormonal control, hence, levels of these enzymes can be affected by exposure to endocrine-disrupting chemicals. Previous research has reported that treatment of fish and other animals with the estrogenic and androgenic hormones 17beta-estradiol (E2) and testosterone (T) alters the P450 content or enzyme activities in the treated animals. However, the results of many of these studies are either incomplete or in disagreement and in most cases the effect on specific P450 forms has not been determined. Therefore, to better understand the effects of gonadal hormones on the expression of P450s and their associated enzyme activities, it was of interest to undertake a comprehensive investigation of the transcriptional and translational expression of three constitutive hepatic P450s in the rainbow trout (Oncorhynchus mykiss) following hormone exposure. Accordingly, juvenile trout were injected intraperitoneally with propylene glycol vehicle and the most active estrogenic and androgenic hormones E2 (3 mg/kg) or T (3 mg/kg) on days 1, 4, 7, 13, and 15 and euthanized on day 19. After treatment with E2, hepatic microsomes showed significantly lower levels (percentage of control) in total P450 contents (52%), lauric acid hydroxylase (32%), and 6beta progesterone hydroxylase activities (27%), [(3)H]aflatoxin-DNA binding (31%), and the protein levels of individual cytochrome P450s (CYPs) LMC1 (CYP2M1), LMC2, (CYP2K1), and LMC5 (CYP3A27) (average for three isoforms a reduction to 29% of control values) with only minor differences between sexes. Treatment with T had either no effect or resulted in small increases in total P450 in males (42%), in lauric acid hydroxylase in females (24%), and in 6beta-progesterone hydroxylase activity in males (21%). Biological variabilities among fish were high and a polymorphic or new LMC2-like form was detected at about 52 kDa in some liver microsomal samples after exposure of fish to either hormone. Female liver RNAs were analyzed through Northern blots and an average decrease of 94% in CYP2 M1, CYP2K1, and CYP3A27 mRNA levels occurred in the E2-treated trout. In livers from T-treated trout, the changes of mRNA levels of CYP2M1 and CYP3A27 were negligible, but CYP2K1 mRNA level decreased by about 60%. Additional CYP2K1 cDNA hybridizable mRNAs were seen in some fish as faint bands at about 2.8 kb for both hormone treatments. Results of this study, therefore, indicated that E2 down regulated while T produced small but variable effects on the hepatic mRNA/protein levels of CYP2K1, CYP2M1, and CYP3A27 in juvenile rainbow trout. This study, therefore, suggests that exposure of fish and other wildlife to environmental endocrine disruptors, especially estrogen mimics, can adversely affect a number of physiological processes through mechanisms involving altered levels of expression of specific P450 isozymes. PMID- 11032765 TI - Organophosphorus compound-induced apoptosis in SH-SY5Y human neuroblastoma cells. AB - Organophosphorus (OP) compounds have been shown to be cytotoxic to SH-SY5Y human neuroblastoma cell cultures. The mechanisms involved in OP compound-induced cell death (apoptosis versus necrosis) were assessed morphologically by looking at nuclear fragmentation and budding using the fluorescent stain Hoechst 33342 (10 microgram/ml). Hoechst staining revealed significant paraoxon (1 mM), parathion (1 mM), phenyl saligenin phosphate (PSP, 10 and 100 microM), tri-ortho-tolyl phosphate (TOTP, 100 microM and 1 mM), and triphenyl phosphite (TPPi, 1 mM) induced time-dependent increases in traditional apoptosis (p < 0.05). In many cells, PSP and TOTP (1 mM) also induced nuclear condensation with little fragmentation or budding. Pretreatment with cyclosporin A (500 nM, 30 h) decreased apoptosis following 1 mM parathion and TOTP exposures. Apoptotic nuclear changes were verified by DNA gel electrophoresis. Activation of caspase 3, a cysteine aspartate protease, was also monitored. OP compounds induced significant time-dependent increases in caspase-3 activation following paraoxon (1 mM), parathion (100 microM, 1 mM), PSP (10 microM, 100 microM, 1 mM), TOTP (100 microM, 1 mM), and TPPi (1 mM) exposure (p < 0.05). Pretreatment with cyclosporin A (500 nM, 30 h) significantly decreased caspase-3 activation during extended incubations with paraoxon, parathion, and TPPi (p < 0.05). In addition, pretreatment with the caspase-3 inhibitor Ac-DEVD-CHO and the caspase-8 inhibitor Ac-IETD-CHO (25 microM, 8 h) significantly decreased caspase-3 activation following exposure to 1 mM PSP and parathion (p < 0.05). Pretreatment with the serine protease inhibitor phenylmethyl sulfonyl fluoride (PMSF; 1 mM, 8 h) also significantly decreased caspase activation following 1 mM PSP and TOTP exposures (p < 0.05). Alteration of OP compound-induced nuclear fragmentation or caspase-3 activation by pretreatment with cyclosporin A, Ac-IETD-CHO, or PMSF suggested that OP compound-induced cytotoxicity may be modulated through multiple sites, including mitochondrial permeability pores, receptor-mediated caspase pathways, or serine proteases. PMID- 11032766 TI - Short-term treatment with alcohols causes hepatic steatosis and enhances acetaminophen hepatotoxicity in Cyp2e1(-/-) mice. AB - CYP2E1 has been reported to have an essential role in alcohol-mediated increases in hepatic steatosis and acetaminophen hepatotoxicity. We found that pretreatment of Cyp2e1(-/-) mice with ethanol plus isopentanol, the predominant alcohols in alcoholic beverages, for 7 days resulted in micro- and macrovesicular steatosis in the livers of all mice, as well as a dramatic increase in acetaminophen hepatotoxicity. In Cyp2e1(-/-) mice administered up to 600 mg acetaminophen/kg alone and euthanized 7 h later, there was no increase in serum levels of ALT. In Cyp2e1(-/-) mice pretreated with ethanol and isopentanol, subsequent exposure to 400 or 600 mg acetaminophen/kg resulted in centrilobular necrosis in all mice with maximal elevation in serum levels of ALT. Acetaminophen-mediated liver damage was similar in males and females. Hepatic microsomal levels of APAP activation in untreated females were similar to those in males treated with the alcohols. However, the females, like the males, required pretreatment with the alcohols in order to increase APAP hepatotoxicity. These findings suggest that, in the Cyp2e1(-/-) mice, the alcohol-mediated increase in acetaminophen hepatotoxicity involves the contribution of other factors, in addition to induction of CYP(s) that activate acetaminophen. Alternatively, CYP-mediated activation of acetaminophen measured in vitro may not reflect the actual activity in vivo. Our findings that a 7-day treatment with ethanol and isopentanol causes extensive hepatic steatosis and increases acetaminophen hepatotoxicity in Cyp2e( /-) mice indicate that CYP2E1 is not essential for either response. PMID- 11032767 TI - Effects of captopril on interleukin-6, leukotriene B(4), and oxidative stress markers in serum and inflammatory exudate of arthritic rats: evidence of antiinflammatory activity. AB - We previously demonstrated that captopril (CP) exhibited a high ability to inhibit enzymatically generated leukotrienes, particularly LTB(4), from stimulated intact human neutrophils. This finding together with the immunosuppressive effect of CP have proposed a possible antiinflammatory activity for the drug. Thus, the present study was conducted to investigate the effect of CP on immunologically mediated chronic inflammation; two models were chosen, namely, Freund's adjuvant arthritis and mixed-type hypersensitivity in rat. The effect of CP was assessed on the basis of physical parameter (paw edema) and biochemical markers in blood and inflammatory exudate. CP was given daily during the course of inflammation development. It was administered ip at three doses, viz. 1, 10, and 100 mg/kg. The results claimed that CP succeeded in suppressing edema evolution in hind paws of Freund's arthritic animals, during all phases of the disease. During the chronic phase of inflammation, in either model, CP reduced the elevated serum and exudate (local) LTB(4) and IL-6 levels. The effect on LTB(4) was more pronounced in the exudate and tended to be dose-related. The antiarthritic effect of CP was also accompanied by augmentation of serum level of protein thiols, with reduction or normalization of elevated systemic and/or local levels of lipid peroxide, superoxide dismutase, and glutathione. It could be concluded that long-term treatment with CP confers a good antiinflammatory activity against arthritis in rat, leading to improvement of the oxidative stress induced by the arthritic insult. The reparative effect of the drug could be mediated via reduction of LTB(4) and IL-6. PMID- 11032768 TI - Diesel exhaust, carbon black, and silica particles display distinct Th1/Th2 modulating activity. AB - Certain particulate air pollutants may play an important role in the increasing prevalence of respiratory allergy by stimulating T helper 2 cell (Th2)-mediated immune responses to common antigens. The study described here examined different particles, diesel exhaust particles (DEP), carbon black particles (CBP), and silica particles (SIP) for their immunomodulating capacity in both primary and secondary immune responses in female BALB/C mice. The primary response was studied after subcutaneous injection of 1 mg of particle together with 10 microgram of reporter antigen TNP-OVA (2,4,6-trinitrophenyl coupled to ovalbumin) into the hind paw. Interferon-gamma (IFN-gamma) and interleukin 4 (IL-4) production was assessed in the popliteal lymph node (PLN) at Day 2 and Day 5 after injection by flow cytometry and ELISA. The number of IL-4-containing CD4(+) T cells increased between Day 2 and Day 5 in DEP- and CBP-exposed mice, in contrast to SIP-treated animals. IL-4 production by cultured PLN cells was also significantly increased for DEP- and CBP-treated animals. The secondary response was studied in different organs after an intranasal challenge with TNP-OVA (50 microgram), which was given 4 weeks after the initial subcutaneous injection. Five days after challenge the number of antibody-forming cells (AFCs) was assessed in peribronchial lymph nodes (PBLN), spleen, bone marrow, and PLN, and antibody levels were determined in weekly obtained blood samples. It appeared that all particles acted as adjuvant, but the different particles stimulated distinct types of immune responses to TNP-OVA. DEP-treated animals show high IgG1 and IgE levels in serum and high IgG1 and IgE-forming AFC numbers in PBLN, bone marrow, and spleen. CBP-treated animals show even higher IgG1 and IgE levels and AFC numbers, and in addition display IgG2a production. SIP-injected animals display predominantly IgG2a responses. It is concluded that DEP are able to skew the immune response toward the T helper 2 (Th2) side, whereas SIP stimulate a Th1 response and CBP have a mixed activity, stimulating both Th1 and Th2 responses in this model. PMID- 11032769 TI - Effect of ozone on diesel exhaust particle toxicity in rat lung. AB - Ambient particulate matter (PM) concentrations have been associated with mortality and morbidity. Diesel exhaust particles (DEP) are present in ambient urban air PM. Coexisting with DEP (and PM) is ozone (O(3)), which has the potential to react with some components of DEP. Some reports have shown increased lung injury in rats coexposed to PM and O(3), but it is unclear whether this increased injury was due to direct interaction between the pollutants or via other mechanisms. To examine whether O(3) can directly react with and affect PM bioactivity, we exposed DEP to O(3) in a cell-free in vitro system and then examined the bioactivity of the resultant DEP in a rat model of lung injury. Standard Reference Material 2975 (diesel exhaust PM) was initially exposed to 0.1 ppm O(3) for 48 h and then instilled intratracheally in Sprague-Dawley rats. Rat lung inflammation and injury was examined 24 h after instillation by lung lavage. The DEP exposed to 0.1 ppm O(3) was more potent in increasing neutrophilia, lavage total protein, and LDH activity compared to unexposed DEP. The increased DEP activity induced by the O(3) exposure was not attributable to alteration by air that was also present during the O(3) exposure. Exposure of DEP to a higher O(3) concentration (1.0 ppm) led to a decreased bioactivity of the particles. In contrast, carbon black particles, low in organic content relative to DEP, did not exhibit an increase in any of the bioactivities examined after exposure to 0.1 ppm O(3). DEP incorporated O(3) (labeled with (18)O) in a linear fashion. These data suggest that ambient concentrations of O(3) can increase the biological potency of DEP. The ozonized DEP may play a role in the induction of lung responses by ambient PM. PMID- 11032770 TI - Predicted chemical warfare agent VX toxicity to uniformed soldier using parathion in vitro human skin exposure and absorption. AB - Chemical warfare agents (CWA) are easily and inexpensively produced and are a significant threat to military forces and the public. Most well-known CWAs are organophosphorus compounds, a number or which are used as pesticides, including parathion. This study determined the in vitro percutaneous absorption of parathion as a CWA simulant through naked human skin and uniformed skin (dry and sweated). Parathion percentage dose absorbed through naked skin (1.78 +/- 0. 41) was greater than dry uniformed skin (0.29 +/- 0.17; p = 0.000) and sweated uniformed skin (0.65 +/- 0.16; p = 0.000). Sweated and dry uniformed skin absorption were also different (p = 0.007). These relative dry and sweated uniformed skin absorptions were then applied to VX skin permeability for naked skin (head, neck, arms, and hands) and the remaining uniformed skin over the various regions of the human body. Risk assessment shows VX 50% lethality within the first hour for a soldier wearing a sweated uniform. By 8 h postexposure to naked skin plus trunk area predicted lethality for both dry and sweated uniform, and, at 96 h postexposure, all body regions individually exposed would produce lethality. Military uniform and public clothing provide some immediate protection but absorption through cloth and skin does occur. Immediate safety response to skin and clothing is required. PMID- 11032771 TI - Regulation of gamma-glutamate-cysteine ligase expression by oxidative stress in the mouse preimplantation embryo. AB - The present study examined expression of gamma-glutamate-cysteine ligase (GLCL; also known as gamma-glutamylcysteine synthetase), the rate-limiting enzyme for de novo synthesis of glutathione, in the preimplantation mouse embryo. Previous studies indicated that the cleavage stage embryo is unable to synthesize glutathione de novo. It is hypothesized that GLCL mRNA and protein are not normally expressed in the cleavage stage embryo, but either glutathione depletion or oxidation may induce their expression. In untreated embryos, RT-PCR and Western blotting revealed GLCL heavy subunit (GLCL-H) mRNA and protein only at the blastocyst stage of development. Furthermore, while diethyl maleate (DEM) exposure to deplete cellular glutathione did not induce expression of GLCL-H, exposure to tertiary-butyl hydroperoxide (tBH), an oxidizing agent, resulted in significant upregulation of GLCL-H expression in two-cell embryos. Neither treatment affected expression in blastocysts. Finally, HPLC analysis confirmed that tBH-treated embryos experienced oxidative stress, as indicated by an increase in the ratio of oxidized to reduced glutathione. This oxidative stress induced de novo glutathione synthesis in the cleavage stage embryo, as demonstrated by the subsequent recovery of reduced glutathione levels following DEM-induced depletion. In the absence of tBH treatment, however, cleavage stage embryos could not recover GSH after DEM-mediated depletion. This study demonstrates that the preimplantation embryo has the capacity to upregulate glutathione synthesis in response to oxidative stress but not GSH depletion. These results suggest that, while the preimplantation embryo is well adapted to dealing with oxidative stress, it may be poorly protected from GSH-depleting toxicants. PMID- 11032772 TI - Relative contributions of affinity and intrinsic efficacy to aryl hydrocarbon receptor ligand potency. AB - Models of receptor action are valuable for describing properties of ligand receptor interactions and thereby contribute to mechanism-based risk assessment of receptor-mediated toxic effects. In order to build such a model for the aryl hydrocarbon receptor (AHR), binding affinities and CYP1A induction potencies were measured in PLHC-1 cells and were used to determine intrinsic efficacies for 10 halogenated aromatic hydrocarbons (HAH): 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7, 8-tetrachlorodibenzofuran (TCDF), and eight polychlorinated biphenyls (PCB). TCDD, TCDF, and non-ortho-substituted PCBs 77, 81, 126, and 169 behaved as full agonists and displayed high-intrinsic efficacy. In contrast, the mono- and di-ortho-substituted PCBs bound to the AHR but displayed lower or no intrinsic efficacy. PCB 156 was a full agonist, but with an intrinsic efficacy 10 to 50-fold lower than non-ortho-substituted PCBs. PCB 118 was a very weak partial agonist. PCBs 105 and 128 were shown to be competitive antagonists in this system. The model was then used to predict CYP1A induction by binary mixtures. These predictions were tested with binary mixtures of PCB 126, 128, or 156 with TCDD. Both PCB 156 (a low-intrinsic efficacy agonist) and PCB 128 (a competitive antagonist) inhibited the response to TCDD, while the response to TCDD and PCB126 was additive. These data support the following conclusions: 1) only 1-2% of the receptors in the cell need be occupied to achieve 50% of maximal CYP1A induction by one of the high-intrinsic efficacy agonists, demonstrating the existence of "spare" receptors in this system; 2) the insensitivity of fish to ortho-substituted PCBs is due to both reduced affinity and reduced intrinsic efficacy compared to non-ortho-substituted PCBs; 3) PCB congeners exhibit distinct structure-affinity and structure-efficacy relationships. Separation of AHR ligand action into the properties of affinity and intrinsic efficacy allows for improved prediction of the behavior of complex mixtures of ligands, as well as mechanistic comparisons across species and toxic endpoints. PMID- 11032773 TI - Toxicity of cobalt-containing dusts. PMID- 11032774 TI - Reply PMID- 11032775 TI - DBP exerts its antiandrogenic activity by indirectly interfering with androgen signaling pathways. PMID- 11032776 TI - Reply PMID- 11032777 TI - A comparison of fluorescent Ca2+ indicator properties and their use in measuring elementary and global Ca2+ signals. AB - Quantifying the magnitude of Ca2+ signals from changes in the emission of fluorescent indicators relies on assumptions about the indicator behaviour in situ. Factors such as osmolarity, pH, ionic strength and protein environment can affect indicator properties making it advantageous to calibrate indicators within the required cellular or subcellular environment. Selecting Ca2+ indicators appropriate for a particular application depends upon several considerations including Ca2+ binding affinity, dynamic range and ease of loading. These factors are usually best determined empirically. This study describes the in-situ calibration of a number of frequently used fluorescent Ca2+ indicators (Fluo-3, Fluo-4, Calcium Green-1, Calcium Orange, Oregon Green 488 BAPTA-1 and Fura-Red) and their use in reporting low- and high-amplitude Ca2+ signals in HeLa cells. All Ca2+ indicators exhibited lower in-situ Ca2+ binding affinities than suggested by previously published in-vitro determinations. Furthermore, for some of the indicators, there were significant differences in the apparent Ca2+ binding affinities between nuclear and cytoplasmic compartments. Variation between indicators was also found in their dynamic ranges, compartmentalization, leakage and photostability. Overall, Fluo-3 proved to be the generally most applicable Ca2+ indicator, since it displayed a large dynamic range, low compartmentalization and an appropriate apparent Ca2+ binding affinity. However, it was more susceptible to photobleaching than many of the other Ca2+ indicators. PMID- 11032778 TI - Interference of heavy metal cations with fluorescent Ca2+ probes does not affect Ca2+ measurements in living cells. AB - In studies about the effects of heavy metals on intracellular Ca2+, the use of fluorescent probes is debated, as metal cations are known to affect the probe signal. In this study, spectrofluorimetric experiments in free solution, using Fluo-3 and Fura-2, showed that Zn2+ and Cd2+ enhanced the probe signal, Cu2+ quenched it, and Hg2+ had no effect. Addition of GSH prevented most of these effects, suggesting the occurrence of a similar protective role in living cells. Digital imaging of living mussel haemocytes loaded with Fura-2/AM or Fluo-3/AM showed that Hg2+, Cu2+ and Cd2+ induced a rise in probe fluorescence, whereas up to 200 microM Zn2+ had no effect. In particular, Cd2+ produced the strongest probe signal rise in free solution, but the lowest fluorescence increase in cells. Probe calibration yielded [Ca2+]i values characteristic of resting levels in control and Zn2+-exposed cells, and, as expected, indicated Ca2+ homeostasis impairment in cells exposed to Cd2+, Cu2+ and Hg2+. Our results show that Ca2+ probe responses to heavy metals in living cells are completely different from those obtained in free solution, indicating that fluorescent probes can be a suitable tool to record the effects of heavy metals on [Ca2+]i. PMID- 11032779 TI - The spatial relationship between Ca2+ channels and Ca2+-activated channels and the function of Ca2+-buffering in avian sensory neurons. AB - In order to learn about the endogenous Ca2+-buffering in the cytoplasm of chick dorsal root ganglion (DRG) neurons and the distance separating the ryanodine receptor Ca2+ release channels (RyRs) from the plasma membrane, we monitored the amplitude and time course of Ca2+-activated Cl- currents (I(ClCa)) in protocols that manipulated Ca2+-buffering. I(ClCa)was activated by Ca2+ influx via voltage gated Ca2+ channels or by Ca2+ release via RyRs activated by 10 mM caffeine. I(ClCa)was measured in neurons at 20 degrees C and 35 degrees C using the amphotericin perforated patch technique that preserves endogenous Ca2+-buffering, or at 20 degrees C in neurons dialyzed with pipette solutions designed to replace the endogenous Ca2+ buffers. The amplitude of I(ClCa)activated by Ca2+ influx or Ca2+ at 20 degrees C was similar in the amphotericin neurons and neurons dialyzed with an 'unbuffered' pipette solution containing 10 mM citrate and 3 mM ATP as the only Ca2+ binding molecules. Thus, endogenous mobile Ca2+ buffers are relatively unimportant in chick DRG neurons. Warming the neurons from 20 degrees C to 35 degrees C increased the amplitude and the rate of deactivation of I(ClCa)consistent with an increased rate of Ca2+ buffering by fixed endogenous Ca2+-buffers. Dialysis with 2 mM EGTA/0.1 microM free Ca2+ reduced the amplitude and increased the rate of deactivation of I(ClCa)activated by Ca2+ influx and abolished I(ClCa)activated by Ca2+ release. Dialysis with 2 mM BAPTA/0.1 microM free Ca2+ abolished I(ClCa)activated by Ca2+ influx or release. Dialysis with 42 mM HEEDTA/0.5 microM free Ca2+ caused the persistent activation of I(ClCa). Calculations using a Ca2+-diffusion model suggest that the voltage-gated Ca2+ channels and the Ca2+-activated Cl- channels are separated by 50-400 nm and that the RyRs are more than 600 nm from the plasma membrane. PMID- 11032780 TI - A novel barium-sensitive calcium influx into rat astrocytes at low external potassium. AB - Cultured rat cerebellar astrocytes, loaded with the Ca2+-sensitive fluorescent dyes Fura-2 or Fluo-3, responded with cytoplasmic Ca2+ transients, when the external K+ concentration was reduced from 5 mM to below 1 mM. Ca2+ transients were generated after changing to a saline containing 0.2 mM K+ in 82% of the cells (n =303) with a delay of up to 4 min. Cultured rat cortical neurones, which responded in high-K+ saline (50 mM) with Ca2+ transients, showed no Ca2+ responses in low K+ (n =22). In acute rat hippocampal brain slices, presumed glial cells responded with Ca2+ transients in low K+ similar to astrocytes in culture (88%, n =17). The Ca2+ transients were observed both in somatic and dendritic regions of cultured astrocytes, as examined with confocal laser scanning microscopy. Patch-clamped astrocytes hyperpolarized in 0.2 mM K+ from an average resting potential of -65 +/- 4 mV to -98 +/- 20 mV (n =15). The Ca2+ transients in low K+ were suppressed in Ca2+-free saline, buffered with 0.5 mM EGTA, but not after depletion of intracellular Ca2+ stores by thapsigargin, cyclopiazonic acid or by Ruthenium Red, indicating that they were due to Ca2+ influx into the cells, and not caused by intracellular Ca2+ release. The addition of different divalent cations revealed that Ba2+, but not Ni2+, Cd2+, Sr2+ or Mg2+, reversibly blocked the Ca2+ transients in low K+. There was a significant reduction of the Ca2+ responses at micromolar Ba2+ concentrations (Ki = 3.8 microM). The application of different K+ channel blockers, tetraethylammonium, dequalinium, tolbutamide, clotrimazole, or quinidine had no effect on the Ca2+ responses. Removal of external Na+, or intracellular acidification by the addition of 40 mM propionate to the saline, had also no influence on the generation of the Ca2+ transients. The results suggest that reducing the external K+ concentration elicits a Ca2+ influx into rat astrocytes which is highly sensitive to Ba2+. It is discussed that this Ca2+ influx might occur through K+ inward rectifier channels, which become Ca2+-permeable when the extracellular K+ concentration decreases to 1 mM or below. PMID- 11032781 TI - Anion channels modulate store-operated calcium influx in human microglia. AB - Recent work from this laboratory has demonstrated that purinergic-mediated depolarization of human microglia inhibited a store-operated pathway for entry of Ca2+. We have used Fura-2 spectrofluorometry to investigate the effects on store operated Ca2+ influx induced by replacement of NaCl with Na-gluconate in extracellular solutions. Three separate procedures were used to activate store operated channels. Platelet activating factor (PAF) was used to generate a sustained influx of Ca2+ in standard physiological saline solution (PSS). The magnitude of this response was depressed by 70% after replacement of PSS with low Cl- PSS. A second procedure used ATP, initially applied in Ca2+-free PSS solution to deplete intracellular stores. The subsequent perfusion of PSS solution containing Ca2+ resulted in a large and sustained entry of Ca2+, which was inhibited by 75% with low Cl- PSS. The SERCA inhibitor cyclopiazonic acid (CPA) was used to directly deplete stores in zero-Ca2+ PSS. Following the introduction of PSS containing Ca2+, a maintained stores-operated influx of Ca2+ was evident which was inhibited by 77% in the presence of the low Cl- PSS. Ca2+ influx was linearly reduced with cell depolarization in elevated K+ (7.5 to 35 mM) suggesting that changes in external Cl- were manifest as altered electrical driving force for Ca2+ entry. However, 50 mM external KCl effectively eliminated divalent entry which may indicate inactivation of this pathway with high magnitudes of depolarization. Patch clamp studies showed low Cl-PSS to cause depolarizing shifts in both holding currents and reversal potentials of currents activated with voltage ramps. The results demonstrate that Cl- channels play an important role in regulating store-operated entry of Ca2+ in human microglia. PMID- 11032782 TI - The effects of low concentrations of caffeine on spontaneous Ca release in isolated rat ventricular myocytes. AB - We have investigated the effects on spontaneous SR Ca release of modulating the sarcoplasmic reticulum ryanodine receptor (RyR) with low (<0.5 mM) concentrations of caffeine. Experiments were performed on isolated rat ventricular myocytes. Intracellular Ca concentration was measured with Indo-1 or Fluo-3 in voltage clamped cells. Spontaneous Ca release was produced by elevating external Ca to 5 mM. Caffeine application increased the frequency of spontaneous release. Both the magnitude of the spontaneous Ca transients and the integral of the resulting Na Ca exchange current were decreased by caffeine. The combination of increased frequency of spontaneous release and decreased Ca efflux per event meant that the Ca efflux per unit time was unaffected by low concentrations of caffeine. The SR Ca content was reduced by caffeine. The extra Ca efflux calculated from the Na-Ca exchange current integrals occurring during the initial burst of spontaneous activity on application of caffeine accounted for this reduction of SR Ca content. In contrast to these maintained effects on spontaneous release, caffeine had only transient effects on stimulated Ca release produced by depolarizing pulses. We conclude that stimulation of the RyR results in spontaneous release at SR Ca contents lower than those at which release would normally occur. Therefore, the balance between normal and spontaneous Ca release can be shifted by modulation of the RyR. This will have important consequences for arrhythmogenesis due to spontaneous Ca release. PMID- 11032783 TI - The Finland-United States investigation of non-insulin-dependent diabetes mellitus genetics (FUSION) study. I. An autosomal genome scan for genes that predispose to type 2 diabetes. AB - We performed a genome scan at an average resolution of 8 cM in 719 Finnish sib pairs with type 2 diabetes. Our strongest results are for chromosome 20, where we observe a weighted maximum LOD score (MLS) of 2.15 at map position 69.5 cM from pter and secondary weighted LOD-score peaks of 2.04 at 56.5 cM and 1.99 at 17.5 cM. Our next largest MLS is for chromosome 11 (MLS = 1.75 at 84.0 cM), followed by chromosomes 2 (MLS = 0.87 at 5.5 cM), 10 (MLS = 0.77 at 75.0 cM), and 6 (MLS = 0.61 at 112.5 cM), all under an additive model. When we condition on chromosome 2 at 8.5 cM, the MLS for chromosome 20 increases to 5.50 at 69.0 cM (P=.0014). An ordered-subsets analysis based on families with high or low diabetes-related quantitative traits yielded results that support the possible existence of disease-predisposing genes on chromosomes 6 and 10. Genomewide linkage disequilibrium analysis using microsatellite marker data revealed strong evidence of association for D22S423 (P=.00007). Further analyses are being carried out to confirm and to refine the location of these putative diabetes-predisposing genes. PMID- 11032784 TI - The Finland-United States investigation of non-insulin-dependent diabetes mellitus genetics (FUSION) study. II. An autosomal genome scan for diabetes related quantitative-trait loci. AB - Type 2 diabetes mellitus is a complex disorder encompassing multiple metabolic defects. We report results from an autosomal genome scan for type 2 diabetes related quantitative traits in 580 Finnish families ascertained for an affected sibling pair and analyzed by the variance components-based quantitative-trait locus (QTL) linkage approach. We analyzed diabetic and nondiabetic subjects separately, because of the possible impact of disease on the traits of interest. In diabetic individuals, our strongest results were observed on chromosomes 3 (fasting C-peptide/glucose: maximum LOD score [MLS] = 3.13 at 53.0 cM) and 13 (body-mass index: MLS = 3.28 at 5.0 cM). In nondiabetic individuals, the strongest results were observed on chromosomes 10 (acute insulin response: MLS = 3.11 at 21.0 cM), 13 (2-h insulin: MLS = 2.86 at 65.5 cM), and 17 (fasting insulin/glucose ratio: MLS = 3.20 at 9.0 cM). In several cases, there was evidence for overlapping signals between diabetic and nondiabetic individuals; therefore we performed joint analyses. In these joint analyses, we observed strong signals for chromosomes 3 (body-mass index: MLS = 3.43 at 59.5 cM), 17 (empirical insulin-resistance index: MLS = 3.61 at 0.0 cM), and 19 (empirical insulin-resistance index: MLS = 2.80 at 74.5 cM). Integrating genome-scan results from the companion article by Ghosh et al., we identify several regions that may harbor susceptibility genes for type 2 diabetes in the Finnish population. PMID- 11032786 TI - Improved inference of relationship for pairs of individuals. AB - Linkage analyses of genetic diseases and quantitative traits generally are performed using family data. These studies assume the relationships between individuals within families are known correctly. Misclassification of relationships can lead to reduced or inappropriately increased evidence for linkage. Boehnke and Cox (1997) presented a likelihood-based method to infer the most likely relationship of a pair of putative sibs. Here, we modify this method to consider all possible pairs of individuals in the sample, to test for additional relationships, to allow explicitly for genotyping error, and to include X-linked data. Using autosomal genome scan data, our method has excellent power to differentiate monozygotic twins, full sibs, parent-offspring pairs, second-degree (2 degrees ) relatives, first cousins, and unrelated pairs but is unable to distinguish accurately among the 2 degrees relationships of half sibs, avuncular pairs, and grandparent-grandchild pairs. Inclusion of X-linked data improves our ability to distinguish certain types of 2 degrees relationships. Our method also models genotyping error successfully, to judge by the recovery of MZ twins and parent-offspring pairs that are otherwise misclassified when error exists. We have included these extensions in the latest version of our computer program RELPAIR and have applied the program to data from the Finland-United States Investigation of Non-Insulin-Dependent Diabetes Mellitus (FUSION) study. PMID- 11032785 TI - Linkage disequilibrium analysis of biallelic DNA markers, human quantitative trait loci, and threshold-defined case and control subjects. AB - Linkage disequilibrium (LD) mapping has been applied to many simple, monogenic, overtly Mendelian human traits, with great success. However, extensions and applications of LD mapping approaches to more complex human quantitative traits have not been straightforward. In this article, we consider the analysis of biallelic DNA marker loci and human quantitative trait loci in settings that involve sampling individuals from opposite ends of the trait distribution. The purpose of this sampling strategy is to enrich samples for individuals likely to possess (and not possess) trait-influencing alleles. Simple statistical models for detecting LD between a trait-influencing allele and neighboring marker alleles are derived that make use of this sampling scheme. The power of the proposed method is investigated analytically for some hypothetical gene-effect scenarios. Our studies indicate that LD mapping of loci influencing human quantitative trait variation should be possible in certain settings. Finally, we consider possible extensions of the proposed methods, as well as areas for further consideration and improvement. PMID- 11032787 TI - Performance of Markov chain-Monte Carlo approaches for mapping genes in oligogenic models with an unknown number of loci. AB - Markov chain-Monte Carlo (MCMC) techniques for multipoint mapping of quantitative trait loci have been developed on nuclear-family and extended-pedigree data. These methods are based on repeated sampling-peeling and gene dropping of genotype vectors and random sampling of each of the model parameters from their full conditional distributions, given phenotypes, markers, and other model parameters. We further refine such approaches by improving the efficiency of the marker haplotype-updating algorithm and by adopting a new proposal for adding loci. Incorporating these refinements, we have performed an extensive simulation study on simulated nuclear-family data, varying the number of trait loci, family size, displacement, and other segregation parameters. Our simulation studies show that our MCMC algorithm identifies the locations of the true trait loci and estimates their segregation parameters well-provided that the total number of sibship pairs in the pedigree data is reasonably large, heritability of each individual trait locus is not too low, and the loci are not too close together. Our MCMC algorithm was shown to be significantly more efficient than LOKI (Heath 1997) in our simulation study using nuclear-family data. PMID- 11032788 TI - Tracing European founder lineages in the Near Eastern mtDNA pool. AB - Founder analysis is a method for analysis of nonrecombining DNA sequence data, with the aim of identification and dating of migrations into new territory. The method picks out founder sequence types in potential source populations and dates lineage clusters deriving from them in the settlement zone of interest. Here, using mtDNA, we apply the approach to the colonization of Europe, to estimate the proportion of modern lineages whose ancestors arrived during each major phase of settlement. To estimate the Palaeolithic and Neolithic contributions to European mtDNA diversity more accurately than was previously achievable, we have now extended the Near Eastern, European, and northern-Caucasus databases to 1,234, 2, 804, and 208 samples, respectively. Both back-migration into the source population and recurrent mutation in the source and derived populations represent major obstacles to this approach. We have developed phylogenetic criteria to take account of both these factors, and we suggest a way to account for multiple dispersals of common sequence types. We conclude that (i) there has been substantial back-migration into the Near East, (ii) the majority of extant mtDNA lineages entered Europe in several waves during the Upper Palaeolithic, (iii) there was a founder effect or bottleneck associated with the Last Glacial Maximum, 20,000 years ago, from which derives the largest fraction of surviving lineages, and (iv) the immigrant Neolithic component is likely to comprise less than one-quarter of the mtDNA pool of modern Europeans. PMID- 11032789 TI - Autosomal, mtDNA, and Y-chromosome diversity in Amerinds: pre- and post-Columbian patterns of gene flow in South America. AB - To evaluate sex-specific differences in gene flow between Native American populations from South America and between those populations and recent immigrants to the New World, we examined the genetic diversity at uni- and biparental genetic markers of five Native American populations from Colombia and in published surveys from native South Americans. The Colombian populations were typed for five polymorphisms in mtDNA, five restriction sites in the beta-globin gene cluster, the DQA1 gene, and nine autosomal microsatellites. Elsewhere, we published results for seven Y-chromosome microsatellites in the same populations. Autosomal polymorphisms showed a mean G(ST) of 6.8%, in agreement with extensive classical marker studies of South American populations. MtDNA and Y-chromosome markers resulted in G(ST) values of 0.18 and 0.165, respectively. When only Y chromosomes of confirmed Amerind origin were used in the calculations (as defined by the presence of allele T at locus DYS199), G(ST) increased to 0.22. G(ST) values calculated from published data for other South American natives were 0.3 and 0.29 for mtDNA and Amerind Y chromosomes, respectively. The concordance of these estimates does not support an important difference in migration rates between the sexes throughout the history of South Amerinds. Admixture analysis of the Colombian populations suggests an asymmetric pattern of mating involving mostly immigrant men and native women. PMID- 11032791 TI - On a randomization procedure. PMID- 11032790 TI - Strong Amerind/white sex bias and a possible Sephardic contribution among the founders of a population in northwest Colombia. AB - Historical and genetic evidences suggest that the recently founded population of Antioquia (Colombia) is potentially useful for the genetic mapping of complex traits. This population was established in the 16th-17th centuries through the admixture of Amerinds, Europeans, and Africans and grew in relative isolation until the late 19th century. To examine the origin of the founders of Antioquia, we typed 11 markers on the nonrecombining portion of the Y chromosome and four markers on mtDNA in a sample of individuals with confirmed Antioquian ancestry. The polymorphisms on the Y chromosome (five biallelic markers and six microsatellites) allow an approximation to the origin of founder men, and those on mtDNA identify the four major founder Native American lineages. These data indicate that approximately 94% of the Y chromosomes are European, 5% are African, and 1% are Amerind. Y-chromosome data are consistent with an origin of founders predominantly in southern Spain but also suggest that a fraction came from northern Iberia and that some possibly had a Sephardic origin. In stark contrast with the Y-chromosome, approximately 90% of the mtDNA gene pool of Antioquia is Amerind, with the frequency of the four Amerind founder lineages being closest to Native Americans currently living in the area. These results indicate a highly asymmetric pattern of mating in early Antioquia, involving mostly immigrant men and local native women. The discordance of our data with blood-group estimates of admixture suggests that the number of founder men was larger than that of women. PMID- 11032793 TI - The promise and pitfalls of telomere region-specific probes. PMID- 11032794 TI - X-ray crystal structure of rabbit N-acetylglucosaminyltransferase I: catalytic mechanism and a new protein superfamily. AB - N:-acetylglucosaminyltransferase I (GnT I) serves as the gateway from oligomannose to hybrid and complex N:-glycans and plays a critical role in mammalian development and possibly all metazoans. We have determined the X-ray crystal structure of the catalytic fragment of GnT I in the absence and presence of bound UDP-GlcNAc/Mn(2+) at 1.5 and 1.8 A resolution, respectively. The structures identify residues critical for substrate binding and catalysis and provide evidence for similarity, at the mechanistic level, to the deglycosylation step of retaining beta-glycosidases. The structuring of a 13 residue loop, resulting from UDP-GlcNAc/Mn(2+) binding, provides an explanation for the ordered sequential 'Bi Bi' kinetics shown by GnT I. Analysis reveals a domain shared with Bacillus subtilis glycosyltransferase SpsA, bovine beta-1,4-galactosyl transferase 1 and Escherichia coli N:-acetylglucosamine-1-phosphate uridyltransferase. The low sequence identity, conserved fold and related functional features shown by this domain define a superfamily whose members probably share a common ancestor. Sequence analysis and protein threading show that the domain is represented in proteins from several glycosyltransferase families. PMID- 11032795 TI - Crystal structure of Streptococcus pneumoniae acyl carrier protein synthase: an essential enzyme in bacterial fatty acid biosynthesis. AB - Acyl carrier protein synthase (AcpS) catalyzes the formation of holo-ACP, which mediates the essential transfer of acyl fatty acid intermediates during the biosynthesis of fatty acids and lipids in the cell. Thus, AcpS plays an important role in bacterial fatty acid and lipid biosynthesis, making it an attractive target for therapeutic intervention. We have determined, for the first time, the crystal structure of the Streptococcus pneumoniae AcpS and AcpS complexed with 3'5'-ADP, a product of AcpS, at 2.0 and 1.9 A resolution, respectively. The crystal structure reveals an alpha/beta fold and shows that AcpS assembles as a tightly packed functional trimer, with a non-crystallographic pseudo-symmetric 3 fold axis, which contains three active sites at the interface between protomers. Only two active sites are occupied by the ligand molecules. Although there is virtually no sequence similarity between the S.pneumoniae AcpS and the Bacillus subtilis Sfp transferase, a striking structural similarity between both enzymes was observed. These data provide a starting point for structure-based drug design efforts towards the identification of AcpS inhibitors with potent antibacterial activity. PMID- 11032796 TI - Structure of the GAF domain, a ubiquitous signaling motif and a new class of cyclic GMP receptor. AB - GAF domains are ubiquitous motifs present in cyclic GMP (cGMP)-regulated cyclic nucleotide phosphodiesterases, certain adenylyl cyclases, the bacterial transcription factor FhlA, and hundreds of other signaling and sensory proteins from all three kingdoms of life. The crystal structure of the Saccharomyces cerevisiae YKG9 protein was determined at 1.9 A resolution. The structure revealed a fold that resembles the PAS domain, another ubiquitous signaling and sensory transducer. YKG9 does not bind cGMP, but the isolated first GAF domain of phosphodiesterase 5 binds with K:(d) = 650 nM. The cGMP binding site of the phosphodiesterase GAF domain was identified by homology modeling and site directed mutagenesis, and consists of conserved Arg, Asn, Lys and Asp residues. The structural and binding studies taken together show that the cGMP binding GAF domains form a new class of cyclic nucleotide receptors distinct from the regulatory domains of cyclic nucleotide-regulated protein kinases and ion channels. PMID- 11032797 TI - Crystal structure of the cell division protein FtsA from Thermotoga maritima. AB - Bacterial cell division requires formation of a septal ring. A key step in septum formation is polymerization of FtsZ. FtsA directly interacts with FtsZ and probably targets other proteins to the septum. We have solved the crystal structure of FtsA from Thermotoga maritima in the apo and ATP-bound form. FtsA consists of two domains with the nucleotide-binding site in the interdomain cleft. Both domains have a common core that is also found in the actin family of proteins. Structurally, FtsA is most homologous to actin and heat-shock cognate protein (Hsc70). An important difference between FtsA and the actin family of proteins is the insertion of a subdomain in FtsA. Movement of this subdomain partially encloses a groove, which could bind the C-terminus of FtsZ. FtsZ is the bacterial homologue of tubulin, and the FtsZ ring is functionally similar to the contractile ring in dividing eukaryotic cells. Elucidation of the crystal structure of FtsA shows that another bacterial protein involved in cytokinesis is structurally related to a eukaryotic cytoskeletal protein involved in cytokinesis. PMID- 11032798 TI - Structural comparison of dimeric Eg5, Neurospora kinesin (Nkin) and Ncd head-Nkin neck chimera with conventional kinesin. AB - Cryo-electron microscopy and 3D image reconstruction of microtubules saturated with kinesin dimers has shown one head bound to tubulin, the other free. The free head of rat kinesin sits on the top right of the bound head (with the microtubule oriented plus-end upwards) in the presence of 5'-adenylylimido-diphosphate (AMPPNP) and on the top left in nucleotide-free solutions. To understand the relevance of this movement, we investigated other dimeric plus-end-directed motors: Neurospora kinesin (Nkin); Eg5, a slow non-processive kinesin; and a chimera of Ncd heads attached to Nkin necks. In the AMPPNP (ATP-like) state, all dimers have the free head to the top right. In the absence of nucleotide, the free head of an Nkin dimer appears to occupy alternative positions to either side of the bound head. Despite having the Nkin neck, the free head of the chimera was only seen to the top right of the bound head. Eg5 also has the free head mostly to the top right. We suggest that processive movement may require kinesins to move their heads in alternative ways. PMID- 11032799 TI - In vivo covalent cross-linking of cellular actin by the Vibrio cholerae RTX toxin. AB - Enteric pathogens often export toxins that elicit diarrhea as a part of the etiology of disease, including toxins that affect cytoskeletal structure. Recently, we discovered that the intestinal pathogen Vibrio cholerae elicits rounding of epithelial cells that is dependent upon a gene we designated rtxA. Here we investigate the association of rtxA with the cell-rounding effect. We find that V. cholerae exports a large toxin, RTX (repeats-in-toxin) toxin, to culture supernatant fluids and that this toxin is responsible for cell rounding. Furthermore, we find that cell rounding is not due to necrosis, suggesting that RTX toxin is not a typical member of the RTX family of pore-forming toxins. Rather, RTX toxin causes depolymerization of actin stress fibers and covalent cross-linking of cellular actin into dimers, trimers and higher multimers. This RTX toxin-specific cross-linking occurs in cells previously rounded with cytochalasin D, indicating that G-actin is the toxin target. Although several models explain our observations, our simultaneous detection of actin cross linking and depolymerization points toward a novel mechanism of action for RTX toxin, distinguishing it from all other known toxins. PMID- 11032800 TI - Post-translational hydroxylation at the N-terminus of the prion protein reveals presence of PPII structure in vivo. AB - The transmissible spongiform encephalopathies are characterized by conversion of a host protein, PrP(C) (cellular prion protein), to a protease-resistant isoform, PrP(Sc) (prion protein scrapie isoform). The importance of the highly flexible, N terminal region of PrP has recently become more widely appreciated, particularly the biological activities associated with its metal ion-binding domain and its potential to form a poly(L-proline) II (PPII) helix. Circular dichroism spectroscopy of an N-terminal peptide, PrP(37-53), showed that the PPII helix is formed in aqueous buffer; as it also contains an Xaa-Pro-Gly consensus sequence, it may act as a substrate for the collagen-modifying enzyme prolyl 4-hydroxylase. Direct evidence for this modification was obtained by mass spectrometry and Edman sequencing in recombinant mouse PrP secreted from stably transfected Chinese hamster ovary cells. Almost complete conversion of proline to 4-hydroxyproline occurs specifically at residue Pro44 of this murine protein; the same hydroxylated residue was detected, at lower levels, in PrP(Sc) from the brains of scrapie-infected mice. Cation binding and/or post-translational hydroxylation of this region of PrP may regulate its role in the physiology and pathobiology of the cell. PMID- 11032802 TI - A novel regulatory role of glucose transporter of Escherichia coli: membrane sequestration of a global repressor Mlc. AB - External glucose stimulates transcription of several genes including ptsG encoding IICB(Glc), a membrane component of the phosphotransferase system (PTS), by relieving the negative regulation of a global repressor Mlc in Escherichia coli. We investigate here how glucose modulates Mlc action. The Mlc-mediated repression is eliminated by a ptsI mutation, while Mlc is constitutively active in a ptsG mutant. We show that IICB(Glc)-FLAG interacts physically with Mlc in crude extracts prepared from cells in which IICB(Glc) is supposed to exist as the non-phosphorylated form. The IICB(Glc)-Mlc interaction is no longer observed when IICB(Glc) is phosphorylated. Exogenously added purified Mlc binds to purified IICB(Glc)-FLAG. We also demonstrate that Mlc is associated with membrane when IICB(Glc) is dephosphorylated while it is in the cytoplasm when IICB(Glc) is phosphorylated or absent. We conclude that IICB(Glc) regulates the cellular localization of Mlc, depending on its phosphorylation state, which is determined by the availability of external glucose. Thus, glucose induces the transcription of Mlc-regulated promoters by sequestering Mlc to the membrane through dephosphorylation of IICB(Glc). PMID- 11032801 TI - Targeting of the pro-apoptotic VDAC-like porin (PorB) of Neisseria gonorrhoeae to mitochondria of infected cells. AB - Infection of cell cultures with Neisseria gonorrhoeae results in apoptosis that is mediated by the PorB porin. During the infection process porin translocates from the outer bacterial membrane into host cell membranes where its channel activity is regulated by nucleotide binding and voltage-dependent gating, features that are shared by the mitochondrial voltage-dependent anion channel (VDAC). Here we show that porin is selectively and efficiently transported to mitochondria of infected cells. Prevention of porin translocation also blocked the induction of apoptosis. Mitochondria of cells treated with porin both in vitro and in vivo were depleted of cytochrome c and underwent permeability transition. Overexpression of Bcl-2 blocked porin-induced apoptosis. The release of cytochrome c occurred independently of active caspases but was completely prevented by Bcl-2. Our data suggest that the Neisseria porin can, like its eukaryotic homologue, function at the mitochondrial checkpoint to mediate apoptosis. PMID- 11032803 TI - Signal transduction between a membrane-bound transporter, PtsG, and a soluble transcription factor, Mlc, of Escherichia coli. AB - The global regulator Mlc controls several genes implicated in sugar utilization systems, notably the phosphotransferase system (PTS) genes, ptsG, manXYZ and ptsHI, as well as the malT activator. No specific low molecular weight inducer has been identified that can inactivate Mlc, but its activity appeared to be modulated by transport of glucose via Enzyme IICB(Glc) (PtsG). Here we demonstrate that inactivation of Mlc is achieved by sequestration of Mlc to membranes containing dephosphorylated Enzyme IICB(Glc). We show that Mlc binds specifically to membrane fractions which carry PtsG and that excess Mlc can inhibit Enzyme IICB(Glc) phosphorylation by the general PTS proteins and also Enzyme IICB(Glc)-mediated phosphorylation of alpha-methylglucoside. Binding of Mlc to Enzyme IICB(Glc) in vitro required the IIB domain and the IIC-B junction region. Moreover, we show that these same regions are sufficient for Mlc regulation in vivo, via cross-dephosphorylation of IIB(Glc) during transport of other PTS sugars. The control of Mlc activity by sequestration to a transport protein represents a novel form of signal transduction in gene regulation. PMID- 11032804 TI - The F-box protein Skp2 is a ubiquitylation target of a Cul1-based core ubiquitin ligase complex: evidence for a role of Cul1 in the suppression of Skp2 expression in quiescent fibroblasts. AB - The ubiquitin protein ligase SCF(Skp2) is composed of Skp1, Cul1, Roc1/Rbx1 and the F-box protein Skp2, the substrate-recognition subunit. Levels of Skp2 decrease as cells exit the cell cycle and increase as cells re-enter the cycle. Ectopic expression of Skp2 in quiescent fibroblasts causes mitogen-independent S phase entry. Hence, mechanisms must exist for limiting Skp2 protein expression during the G(0)/G(1) phases. Here we show that Skp2 is degraded by the proteasome in G(0)/G(1) and is stabilized when cells re-enter the cell cycle. Rapid degradation of Skp2 in quiescent cells depends on Skp2 sequences that contribute to Cul1 binding and interference with endogenous Cul1 function in serum-deprived cells induces Skp2 expression. Furthermore, recombinant Cul1-Roc1/Rbx1-Skp1 complexes can catalyse Skp2 ubiquitylation in vitro. These results suggest that degradation of Skp2 in G(0)/G(1) is mediated, at least in part, by an autocatalytic mechanism involving a Skp2-bound Cul1-based core ubiquitin ligase and imply a role for this mechanism in the suppression of SCF(Skp2) ubiquitin protein ligase function during the G(0)/G(1) phases of the cell cycle. PMID- 11032805 TI - A role for Ebi in neuronal cell cycle control. AB - Mutations in ebi were isolated as enhancers of an over-proliferation phenotype generated by elevated E2F/DP activity in the Drosophila eye. ebi alleles also strongly suppress a phenotype caused by the cyclin-dependent kinase inhibitor p21, restoring S phases in the second mitotic wave of the developing eye disk. ebi mutant embryos display ectopic S phases within the peripheral nervous system and central nervous system at a time in development when neuronal precursor cells would normally begin to differentiate. Consistent with this, we find that ebi mutants have a reduced capacity to undergo neuronal differentiation, that Ebi physically interacts with Sina and phyllopod, and that Ebi promotes Ttk88 degradation in vitro and in S2 cells. Ectopic expression of Ttk88 inhibited differentiation in embryos and eye discs; however, this block to differentiation was insufficient to promote S phase entry in either of the situations where ebi mutations gave this effect. We conclude that Ebi has two distinct functions; it promotes the degradation of a repressor of neuronal differentiation (Ttk88), and has a second independent function that limits S phase entry. PMID- 11032806 TI - MEKK2 gene disruption causes loss of cytokine production in response to IgE and c Kit ligand stimulation of ES cell-derived mast cells. AB - Ligation of the high-affinity IgE receptor (FcepsilonRI) or of c-Kit stimulates cytokine production in mast cells. We show that MEK kinase 2 (MEKK2), a MAPK kinase kinase (MAP3K) that regulates the JNK and ERK5 pathways, is required for cytokine production in embryonic stem (ES) cell-derived mast cells (ESMC). Targeted disruption of the MEKK2 or MEKK1 gene was used to abolish expression of the respective kinases in ESMC. Transcription of specific cytokines in response to IgE or c-Kit ligand was markedly reduced in MEKK2(-/-) ESMC relative to wild type ESMC. Cytokine production in MEKK1(-/-) ESMC was similar to that of wild type ESMC, demonstrating the specificity of MEKK2 in signaling cytokine gene regulation. MEKK2(-/-) ESMC also lost receptor-mediated stimulation of JNK. In contrast, JNK activation in response to UV irradiation was normal, showing that MEKK2 is required for receptor signaling but not for cellular stress responses. MEKK2 is the first MAP3K shown to be required for mast cell tyrosine kinase receptor signaling controlling cytokine gene expression. PMID- 11032807 TI - Ephrin-A5 modulates cell adhesion and morphology in an integrin-dependent manner. AB - The ephrins are membrane-tethered ligands for the Eph receptor tyrosine kinases, which play important roles in patterning of the nervous and vascular systems. It is now clear that ephrins are more than just ligands and can also act as signalling-competent receptors, participating in bidirectional signalling. We have recently shown that ephrin-A5 signals within caveola-like domains of the plasma membrane upon engagement with its cognate Eph receptor, leading to increased adhesion of the cells to fibronectin. Here we show that ephrin-A5 controls sequential biological events that are consistent with its role in neuronal guidance. Activation of ephrin-A5 induces an initial change in cell adhesion followed by changes in cell morphology. Both effects are dependent on the activation of beta1 integrin involving members of the Src family of protein tyrosine kinases. The prolonged activation of ERK-1 and ERK-2 is required for the change in cell morphology. Our work suggests a new role for class A ephrins in specifying the affinity of the cells towards various extracellular substrates by regulating integrin function. PMID- 11032808 TI - Steroid-induced androgen receptor-oestradiol receptor beta-Src complex triggers prostate cancer cell proliferation. AB - Treatment of human prostate carcinoma-derived LNCaP cells with androgen or oestradiol triggers simultaneous association of androgen receptor and oestradiol receptor beta with Src, activates the Src/Raf-1/Erk-2 pathway and stimulates cell proliferation. Surprisingly, either androgen or oestradiol action on each of these steps is inhibited by both anti-androgens and anti-oestrogens. Similar findings for oestradiol receptor alpha were observed in MCF-7 or T47D cells stimulated by either oestradiol or androgens. Microinjection of LNCaP, MCF-7 and T47D cells with SrcK(-) abolishes steroid-stimulated S-phase entry. Data from transfected Cos cells confirm and extend the findings from these cells. Hormone stimulated Src interaction with the androgen receptor and oestradiol receptor alpha or beta is detected using glutathione S:-transferase fusion constructs. Src SH2 interacts with phosphotyrosine 537 of oestradiol receptor alpha and the Src SH3 domain with a proline-rich stretch of the androgen receptor. The role of this phosphotyrosine is stressed by its requirement for association of oestradiol receptor alpha with Src and consequent activation of Src in intact Cos cells. PMID- 11032809 TI - MAPK/ERK signaling in activated T cells inhibits CD95/Fas-mediated apoptosis downstream of DISC assembly. AB - When T cells are activated, the expression of the CD95 ligand is elevated, with the purpose of inducing apoptosis in target cells and to later eliminate the activated T cells. We have shown previously that mitogen-activated protein kinase (MAPK or ERK) signaling suppresses CD95-mediated apoptosis in different cellular systems. In this study we examined whether MAPK signaling controls the persistence and CD95-mediated termination of an immune response in activated T cells. Our results show that activation of Jurkat T cells through the T cell receptor immediately suppresses CD95-mediated apoptosis, and that this suppression is mediated by MAPK activation. During the phase of elevated MAPK activity, the activation of caspase-8 and Bid is inhibited, whereas the assembly of a functional death-inducing signaling complex (DISC) is not affected. These results explain the resistance to CD95 responses observed during the early phase of T cell activation and suggest that MAPK-activation deflects DISC signaling from activating caspase-8 and Bid. The physiological relevance of the results was confirmed in activated primary peripheral T cells, in which inhibition of MAPK signaling markedly sensitized the cells to CD95-mediated apoptosis. PMID- 11032810 TI - Activation of B-Raf kinase requires phosphorylation of the conserved residues Thr598 and Ser601. AB - The Raf kinase family serves as a central intermediate to relay signals from Ras to ERK. The precise molecular mechanism for Raf activation is still not fully understood. Here we report that phosphorylation of Thr598 and Ser601, which lie between kinase subdomains VII and VIII, is essential for B-Raf activation by Ras. Substitution of these residues by alanine (B-RafAA) abolished Ras-induced B-Raf activation without altering the association of B-Raf with other signaling proteins. Phosphopeptide mapping and immunoblotting with phospho-specific antibodies confirmed that Thr598 and Ser601 are in vivo phosphorylation sites induced by Ras. Furthermore, replacement of these two sites by acidic residues (B RafED) renders B-Raf constitutively active. Con sistent with these data, B-RafAA and B-RafED exhibited diminished and enhanced ability, respectively, to stimulate ERK activation and Elk-dependent transcription. Moreover, functional studies revealed that B-RafED was able to promote NIH 3T3 cell transformation and PC12 cell differentiation. Since Thr598 and Ser601 are conserved in all Raf family members from Caenorhabditis elegans to mammals, we propose that phosphorylation of these two residues may be a general mechanism for Raf activation. PMID- 11032811 TI - Interaction of the type Ialpha PIPkinase with phospholipase D: a role for the local generation of phosphatidylinositol 4, 5-bisphosphate in the regulation of PLD2 activity. AB - Phosphoinositides are localized in various intracellular compartments and can regulate a number of intracellular functions, such as cytoskeletal dynamics and membrane trafficking. Phospholipase Ds (PLDs) are regulated enzymes that hydrolyse phosphatidylcholine (PtdCho) to generate the putative second messenger phosphatidic acid (PtdOH). In vitro, PLDs have an absolute requirement for higher phosphorylated inositides, such as phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)]. Whether this lipid is able to regulate the activity of PLD in vivo is contentious. To examine this hypothesis we studied the relationship between PLD and an enzyme critical for the intracellular synthesis of PtdIns(4,5)P(2): phosphatidylinositol 4-phosphate 5-kinase alpha (Type Ialpha PIPkinase). We find that both PLD1 and PLD2 interact with the Type Ialpha PIPkinase and that PLD2 activity in vivo can be regulated solely by the expression of this lipid kinase. Moreover, PLD2 is able to recruit the Type Ialpha PIPkinase to its intracellular location. We show that the physiological requirement of PLD enzymes for PtdIns(4,5)P(2) is critical and that PLD2 activity can be regulated solely by the levels of this key intracellular lipid. PMID- 11032812 TI - Direct association of ligand-binding and pore domains in homo- and heterotetrameric inositol 1,4,5-trisphosphate receptors. AB - Inositol 1,4,5-trisphosphate receptors (IP(3)Rs) are a family of intracellular Ca(2+) channels that exist as homo- or heterotetramers. In order to determine whether the N-terminal ligand-binding domain is in close physical proximity to the C-terminal pore domain, we prepared microsomal membranes from COS-7 cells expressing recombinant type I and type III IP(3)R isoforms. Trypsin digestion followed by cross-linking and co-immunoprecipitation of peptide fragments suggested an inter-subunit N- and C-terminal interaction in both homo- and heterotetramers. This observation was further supported by the ability of in vitro translated C-terminal peptides to interact specifically with an N-terminal fusion protein. Using a (45)Ca(2+) flux assay, we provide functional evidence that the ligand-binding domain of one subunit can gate the pore domain of an adjacent subunit. We conclude that common structural motifs are shared between the type I and type III IP(3)Rs and propose that the gating mechanism of IP(3)R Ca(2+) channels involves the association of the N-terminus of one subunit with the C-terminus of an adjacent subunit in both homo- and heterotetrameric complexes. PMID- 11032814 TI - The recruitment of RNA polymerase I on rDNA is mediated by the interaction of the A43 subunit with Rrn3. AB - RNA polymerase I (Pol I) is dedicated to transcription of the large ribosomal DNA (rDNA). The mechanism of Pol I recruitment onto rDNA promoters is poorly understood. Here we present evidence that subunit A43 of Pol I interacts with transcription factor Rrn3: conditional mutations in A43 were found to disrupt the transcriptionally competent Pol I-Rrn3 complex, the two proteins formed a stable complex when co-expressed in Escherichia coli, overexpression of Rrn3 suppressed the mutant phenotype, and A43 and Rrn3 mutants showed synthetic lethality. Consistently, immunoelectron microscopy data showed that A43 and Rrn3 co-localize within the Pol I-Rrn3 complex. Rrn3 has several protein partners: a two-hybrid screen identified the C-terminus of subunit Rrn6 of the core factor as a Rrn3 contact, an interaction supported in vitro by affinity chromatography. Our results suggest that Rrn3 plays a central role in Pol I recruitment to rDNA promoters by bridging the enzyme to the core factor. The existence of mammalian orthologues of A43 and Rrn3 suggests evolutionary conservation of the molecular mechanisms underlying rDNA transcription in eukaryotes. PMID- 11032813 TI - Mammalian achaete-scute and atonal homologs regulate neuronal versus glial fate determination in the central nervous system. AB - Whereas vertebrate achaete-scute complex (as-c) and atonal (ato) homologs are required for neurogenesis, their neuronal determination activities in the central nervous system (CNS) are not yet supported by loss-of-function studies, probably because of genetic redundancy. Here, to address this problem, we generated mice double mutant for the as-c homolog Mash1 and the ato homolog Math3. Whereas in Mash1 or Math3 single mutants neurogenesis is only weakly affected, in the double mutants tectal neurons, two longitudinal columns of hindbrain neurons and retinal bipolar cells were missing and, instead, those cells that normally differentiate into neurons adopted the glial fate. These results indicated that Mash1 and Math3 direct neuronal versus glial fate determination in the CNS and raised the possibility that downregulation of these bHLH genes is one of the mechanisms to initiate gliogenesis. PMID- 11032815 TI - Glycogen synthase kinase-3 enhances nuclear export of a Dictyostelium STAT protein. AB - Extracellular cAMP stimulates the rapid tyrosine phosphorylation and nuclear translocation of the DICTYOSTELIUM: STAT protein Dd-STATa. Here we show that it also induces serine phosphorylation by GskA, a homologue of glycogen synthase kinase-3 (GSK-3). Tyrosine phosphorylation occurs within 10 s of stimulation, whereas serine phosphorylation takes 5 min, matching the kinetics observed for the cAMP regulation of GskA. Phosphorylation by GskA enhances nuclear export of Dd-STATa. The phosphorylated region, however, is not itself a nuclear export signal and we identify a region elsewhere in the protein that mediates nuclear export. These results suggest a biphasic regulation of Dd-STATa, in which extracellular cAMP initially directs nuclear import and then, via GskA, promotes its subsequent export. It also raises the possibility of an analogous regulation of STAT nuclear export in higher eukaryotes. PMID- 11032816 TI - Human DNA ligase I efficiently seals nicks in nucleosomes. AB - The access to DNA within nucleosomes is greatly restricted for most enzymes and trans-acting factors that bind DNA. We report here that human DNA ligase I, which carries out the final step of Okazaki fragment processing and of many DNA repair pathways, can access DNA that is wrapped about the surface of a nucleosome in vitro and carry out its enzymatic function with high efficiency. In addition, we find that ligase activity is not affected by the binding of linker histone (H1) but is greatly influenced by the disposition of the core histone tail domains. These results suggest that the window of opportunity for human DNA ligase I may extend well beyond the first stages of chromatin reassembly after DNA replication or repair. PMID- 11032817 TI - Importin-11, a nuclear import receptor for the ubiquitin-conjugating enzyme, UbcM2. AB - Importins are members of a family of transport receptors (karyopherins) that mediate the nucleocytoplasmic transport of protein and RNA cargoes. We identified importin-11 as a potential new human member of this family, on the basis of limited similarity to the Saccharomyces cerevisiae protein, Lph2p, and cloned the complete open reading frame. Importin-11 interacts with the Ran GTPase, and constitutively shuttles between the nuclear and cytoplasmic compartments. A yeast dihybrid screen identified UbcM2, an E2-type ubiquitin-conjugating enzyme, as a binding partner and potential transport cargo for importin-11. Importin-11 and UbcM2 interact directly, and the complex is disassembled by Ran:GTP but not by Ran:GDP. UbcM2 is constitutively nuclear and shuttles between the nuclear and cytoplasmic compartments. Nuclear import of UbcM2 requires Ran and importin-11, and is inhibited by wheatgerm agglutinin, energy depletion or dominant interfering mutants of Ran and importin-beta. These data establish importin-11 as a new member of the karyopherin family of transport receptors, and identify UbcM2 as a nuclear member of the E2 ubiquitin-conjugating enzyme family. PMID- 11032818 TI - She2p, a novel RNA-binding protein tethers ASH1 mRNA to the Myo4p myosin motor via She3p. AB - RNA localization is a widespread mechanism to achieve localized protein synthesis. In budding yeast, localization of ASH1 mRNA controls daughter cell specific accumulation of the transcriptional regulator Ash1p, which determines mating type switching. ASH1 mRNA localization depends on four independently acting sequences ('zipcodes') within the mRNA. In addition, the class V myosin Myo4p and a set of She proteins with as yet unknown function are essential for ASH1 localization. Here we show that She2p is a novel RNA-binding protein that binds specifically to ASH1 mRNA in vivo and to ASH1 RNA zip codes in vitro. She2p can interact with She3 protein via She3p's C-terminus and becomes localized to the daughter cell tip upon ASH1 expression. The N-terminal coiled-coil domain of She3p is required to form an RNA-independent complex with the heavy chain of the myosin motor protein Myo4p. She2p and She3p are the first examples of adapters for tethering a localized mRNA to the motor protein and might serve as prototypes for RNA-motor protein adapters. PMID- 11032819 TI - Processing of polycistronic guide RNAs is associated with RNA editing complexes in Trypanosoma brucei. AB - In kinetoplastid mitochondrial mRNA editing, post-transcriptional insertion or deletion of uridines is templated by guide RNAs (gRNAs). Pre-mRNAs are encoded by maxicircles, while gRNAs are encoded by both maxicircles and minicircles. We have investigated minicircle transcription and the processing of gRNAs in Trypanosoma brucei. We find that minicircles are transcribed polycistronically and that transcripts are accurately processed by an approximately 19S complex. This gRNA processing activity co-purifies with RNA editing complexes, and both remain associated in 19S complexes. Furthermore, we show that RNA editing complexes associate preferentially with a polycistronic gRNA over non-processed RNAs. We propose that the approximately 19S complexes initially described as RNA editing complex I are gRNA processing complexes that cleave polycistronic gRNA transcripts into monocistrons. PMID- 11032820 TI - Essential role of NAT1/p97/DAP5 in embryonic differentiation and the retinoic acid pathway. AB - NAT1/p97/DAP5 is a newly identified protein that shares homology with the translation initiation factor eIF4G. Studies in vitro and in transfected cells indicated that NAT1 might suppress global translation, thereby repressing cellular proliferation. Here we studied the functions of NAT1 in vivo by disrupting its gene in mice. NAT1(-/-) embryos died during gastrulation, indicating a crucial role for NAT1 in embryogenesis. Undifferentiated NAT1(-/-) embryonic stem cells were normal in morphology, proliferation, global translation and gene expression profile. However, NAT1(-/-) cells exhibited an impaired ability to differentiate: they were resistant to differentiation induced by retinoic acid, and teratomas derived from them consisted of undifferentiated and poorly differentiated tissues. The expression of retinoic acid-responsive genes, such as the cell-cycle inhibitor p21(WAF1), was selectively impaired in NAT1(-/-) cells. Transcription from synthetic retinoic acid-responsive elements was also impaired. These data demonstrated that this translation initiation factor homolog controls specific gene expression pathways required for cellular differentiation. PMID- 11032821 TI - Repair of chromosomal abasic sites in vivo involves at least three different repair pathways. AB - We introduced multiple abasic sites (AP sites) in the chromosome of repair deficient mutants of Escherichia coli, in vivo, by expressing engineered variants of uracil-DNA glycosylase that remove either thymine or cytosine. After introduction of AP sites, deficiencies in base excision repair (BER) or recombination were associated with strongly enhanced cytotoxicity and elevated mutation frequencies, selected as base substitutions giving rifampicin resistance. In these strains, increased fractions of transversions and untargeted mutations were observed. In a recA mutant, deficient in both recombination and translesion DNA synthesis (TLS), multiple AP sites resulted in rapid cell death. Preferential incorporation of dAMP opposite a chromosomal AP site ('A rule') required UmuC. Furthermore, we observed an 'A rule-like' pattern of spontaneous mutations that was also UmuC dependent. The mutation patterns indicate that UmuC is involved in untargeted mutations as well. In a UmuC-deficient background, a preference for dGMP was observed. Spontaneous mutation spectra were generally strongly dependent upon the repair background. In conclusion, BER, recombination and TLS all contribute to the handling of chromosomal AP sites in E.coli in vivo. PMID- 11032822 TI - Role of ERCC1 in removal of long non-homologous tails during targeted homologous recombination. AB - The XpF/Ercc1 structure-specific endonuclease performs the 5' incision in nucleotide excision repair and is the apparent mammalian counterpart of the Rad1/Rad10 endonuclease from Saccharomyces cerevisiae. In yeast, Rad1/Rad10 endonuclease also functions in mitotic recombination. To determine whether XpF/Ercc1 endonuclease has a similar role in mitotic recombination, we targeted the APRT locus in Chinese hamster ovary ERCC1(+) and ERCC1(-) cell lines with insertion vectors having long or short terminal non-homologies flanking each side of a double-strand break. No substantial differences were evident in overall recombination frequencies, in contrast to results from targeting experiments in yeast. However, profound differences were observed in types of APRT(+) recombinants recovered from ERCC1(-) cells using targeting vectors with long terminal non-homologies-almost complete ablation of gap repair and single reciprocal exchange events, and generation of a new class of aberrant insertion/deletion recombinants absent in ERCC1(+) cells. These results represent the first demonstration of a requirement for ERCC1 in targeted homologous recombination in mammalian cells, specifically in removal of long non-homologous tails from invading homologous strands. PMID- 11032823 TI - Species-specific double-strand break repair and genome evolution in plants. AB - Even closely related eukaryotic species may differ drastically in genome size. While insertion of retroelements represents a major source of genome enlargement, the mechanism mediating species- specific deletions is fairly obscure. We analyzed the formation of deletions during double-strand break (DSB) repair in Arabidopsis thaliana and tobacco, two dicotyledonous plant species differing >20 fold in genome size. DSBs were induced by the rare cutting restriction endonuclease I-SCE:I and deletions were identified by loss of function of a negative selectable marker gene containing an I-SCE:I site. Whereas the partial use of micro-homologies in junction formation was similar in both species, in tobacco 40% of the deletions were accompanied by insertions. No insertions could be detected in Arabidopsis , where larger deletions were more frequent, indicating a putative inverse correlation between genome size and the average length of deletions. Such a correlation has been postulated before by a theoretical study on the evolution of related insect genomes and our study now identifies a possible molecular cause for the phenomenon, indicating that species specific differences in DSB repair might indeed influence genome evolution. PMID- 11032824 TI - A dynamically tuned double-stranded RNA binding mechanism for the activation of antiviral kinase PKR. AB - A key step in the activation of interferon-inducible antiviral kinase PKR involves differential binding of viral double-stranded RNA (dsRNA) to its two structurally similar N-terminal dsRNA binding motifs, dsRBM1 and dsRBM2. We show here, using NMR spectroscopy, that dsRBM1 with higher RNA binding activity exhibits significant motional flexibility on a millisecond timescale as compared with dsRBM2 with lower RNA binding activity. We further show that dsRBM2, but not dsRBM1, specifically interacts with the C-terminal kinase domain. These results suggest a dynamically tuned dsRNA binding mechanism for PKR activation, where motionally more flexible dsRBM1 anchors to dsRNA, thereby inducing a cooperative RNA binding for dsRBM2 to expose the kinase domain. PMID- 11032825 TI - Compartmentalization of phage phi29 DNA replication: interaction between the primer terminal protein and the membrane-associated protein p1. AB - The bacteriophage phi29 replication protein p1 (85 amino acids) is membrane associated in Bacillus subtilis-infected cells. The C-terminal 52 amino acid residues of p1 are sufficient for assembly into protofilament sheet structures. Using chemical cross-linking experiments, we demonstrate here that p1DeltaC43, a C-terminally truncated p1 protein that neither associates with membranes in vivo nor self-interacts in vitro, can interact with the primer terminal protein (TP) in vitro. Like protein p1, plasmid-encoded protein p1DeltaC43 reduces the rate of phi29 DNA replication in vivo in a dosage-dependent manner. We also show that truncated p1 proteins that retain the N-terminal 42 amino acids, when present in excess, interfere with the in vitro formation of the TP.dAMP initiation complex in a reaction that depends on the efficient formation of a primer TP-phi29 DNA polymerase heterodimer. This interference is suppressed by increasing the concentration of either primer TP or phi29 DNA polymerase. We propose a model for initiation of in vivo phi29 DNA replication in which the viral replisome attaches to a membrane-associated p1-based structure. PMID- 11032826 TI - Inhibition of the transforming growth factor beta 1 signaling pathway by the AML1/ETO leukemia-associated fusion protein. AB - The t(8;21) translocation, found in adult acute myelogenous leukemia, results in the formation of an AML1/ETO chimeric transcription factor. AML1/ETO expression leads to alterations in hematopoietic progenitor cell differentiation, although its role in leukemic transformation is not clear. The N-terminal portion of AML1, which is retained in AML1/ETO, contains a region of homology to the FAST proteins, which cooperate with Smads to regulate transforming growth factor beta1 (TGF-beta1) target genes. We have demonstrated the physical association of Smad proteins with AML1 and AML1/ETO by immunoprecipitation and have mapped the region of interaction to the runt homology domain in these AML1 proteins. Using confocal microscopy, we demonstrated that AML1, and ETO and/or AML1/ETO, colocalize with Smads in the nucleus of t(8;21)-positive Kasumi-1 cells, in the presence but not the absence of TGF-beta1. Using transient transfection assays and a reporter gene construct that contains both Smad and AML1 consensus binding sequences, we demonstrated that overexpression of AML1B cooperates with TGF-beta1 in stimulating reporter gene activity, whereas AML1/ETO represses basal promoter activity and blocks the response to TGF-beta1. Considering the critical role of TGF-beta1 in the growth and differentiation of hematopoietic cells, interference with TGF-beta1 signaling by AML1/ETO may contribute to leukemogenesis. PMID- 11032827 TI - Peroxisomal degradation of trans-unsaturated fatty acids in the yeast Saccharomyces cerevisiae. AB - Degradation of trans-unsaturated fatty acids was studied in the yeast Saccharomyces cerevisiae. Propagation of yeast cells on trans-9 elaidic acid medium resulted in transcriptional up-regulation of the SPS19 gene, whose promoter contains an oleate response element. This up-regulation depended on the Pip2p-Oaf1p transcription factor and was accompanied by induction of import competent peroxisomes. Utilization of trans fatty acids as a single carbon and energy source was evaluated by monitoring the formation of clear zones around cell growth on turbid media containing fatty acids dispersed with Tween 80. For metabolizing odd-numbered trans double bonds, cells required the beta-oxidation auxiliary enzyme Delta(3)-Delta(2)-enoyl-CoA isomerase Eci1p. Metabolism of the corresponding even-numbered double bonds proceeded in the absence of Sps19p (2,4 dienoyl-CoA reductase) and Dci1p (Delta(3,5)-Delta(2,4)-dienoyl-CoA isomerase). trans-2,trans-4-Dienoyl-CoAs could enter beta-oxidation directly via Fox2p (2 enoyl-CoA hydratase 2 and d-specific 3-hydroxyacyl-CoA dehydrogenase) without the involvement of Sps19p, whereas trans-2,cis-4-dienoyl-CoAs could not. This reductase-independent metabolism of trans-2,trans-4-dienoyl-CoAs resembled the situation postulated for mammalian mitochondria in which oleic acid is degraded through a di-isomerase-dependent pathway. In this hypothetical process, trans 2,trans-4-dienoyl-CoA metabolites are generated by Delta(3)-Delta(2)-enoyl-CoA isomerase and Delta(3,5)-Delta(2,4)-dienoyl-CoA isomerase and are degraded by 2 enoyl-CoA hydratase 1 in the absence of 2,4-dienoyl-CoA reductase. Growth of a yeast fox2sps19Delta mutant in which Fox2p was exchanged with rat peroxisomal multifunctional enzyme type 1 on trans-9,trans-12 linolelaidic acid medium gave credence to this theory. We propose an amendment to the current scheme of the carbon flux through beta-oxidation taking into account the dispensability of beta oxidation auxiliary enzymes for metabolizing trans double bonds at even-numbered positions. PMID- 11032828 TI - p130/E2F4 binds to and represses the cdc2 promoter in response to p53. AB - p53 represses the transcription of cdc2 and cyclin B1, causing loss of Cdc2 activity and G(2) arrest. Here we show that the region -22 to -2 of the cdc2 promoter called the R box is required for repression by p53 but not for basal promoter activity. The R box confers p53-dependent repression on heterologous promoters and binds to p130/E2F4 in response to overexpression of p53. R box dependent repression requires p21/waf1, and overexpression of p21/waf1 also represses the cdc2 promoter. These observations suggest that p53 represses the cdc2 promoter by inducing p21/waf1, which inhibits cyclin-dependent kinase activity, enhancing the binding of p130 and E2F4, which together bind to and repress the cdc2 promoter. PMID- 11032829 TI - Post-transcriptional regulation of the gli1 oncogene by the expression of alternative 5' untranslated regions. AB - The oncogene GLI1 is involved in the formation of basal cell carcinoma and other tumor types as a result of the aberrant signaling of the Sonic hedgehog-Patched pathway. In this study, we have identified alternative GLI1 transcripts that differ in their 5' untranslated regions (UTRs) and are generated by exon skipping. These are denoted alpha-UTR, beta-UTR, and gamma-UTR according to the number of noncoding exons possessed (three, two, and one, respectively). The alpha- and beta-UTR forms represent the major Gli1 transcripts expressed in mouse tissues, whereas the gamma-UTR is present at relatively low levels but is markedly induced in mouse skin treated with 12-O-tetradecanoylphorbol 13-acetate. Transcripts corresponding to the murine beta and gamma forms were identified in human tissues, but significantly, only the gamma-UTR form was present in basal cell carcinomas and in proliferating cultures of a keratinocyte cell line. Flow cytometry analysis determined that the gamma-UTR variant expresses a heterologous reporter gene 14-23-fold higher than the alpha-UTR and 5-13-fold higher than the beta-UTR in a variety of cell types. Because expression of the gamma-UTR variant correlates with proliferation, consistent with a role for GLI1 in growth promotion, up-regulation of GLI1 expression through skipping of 5' noncoding exons may be an important tumorigenic mechanism. PMID- 11032830 TI - Sterol-modulated glycolipid sorting occurs in niemann-pick C1 late endosomes. AB - The Niemann-Pick C1 (NPC1) protein and endocytosed low density lipoprotein (LDL) derived cholesterol were shown to enrich separate subsets of vesicles containing lysosomal associated membrane protein 2. Localization of Rab7 in the NPC1 containing vesicles and enrichment of lysosomal hydrolases in the cholesterol containing vesicles confirmed that these organelles were late endosomes and lysosomes, respectively. Lysobisphosphatidic acid, a lipid marker of the late endosomal pathway, was found in the cholesterol-enriched lysosomes. Recruitment of NPC1 to Rab7 compartments was stimulated by cellular uptake of cholesterol. The NPC1 compartment was shown to be enriched in glycolipids, and internalization of GalNAcbeta1-4[NeuAcalpha2-3]Galbeta1-4Glcbeta1-1'-ceramide (G(M2)) into endocytic vesicles depends on the presence of NPC1 protein. The glycolipid profiles of the NPC1 compartment could be modulated by LDL uptake and accumulation of lysosomal cholesterol. Expression in cells of biologically active NPC1 protein fused to green fluorescent protein revealed rapidly moving and flexible tubular extensions emanating from the NPC1-containing vesicles. We conclude that the NPC1 compartment is a dynamic, sterol-modulated sorting organelle involved in the trafficking of plasma membrane-derived glycolipids as well as plasma membrane and endocytosed LDL cholesterol. PMID- 11032831 TI - Neoplastic transformation and tumorigenesis associated with sam68 protein deficiency in cultured murine fibroblasts. AB - Sam68 is a multimeric 68-kDa RNA-binding nuclear protein of unknown function that interacts with, and is tyrosine-phosphorylated by, the oncogenic protein Src during mitosis. Random homozygous knock-out (RHKO) is a retroviral-based antisense RNA strategy that can identify chromosomal genes whose functional disablement leads to reversible tumorigenic capabilities. Here we report that RHKO-induced Sam68 deficiency results in neoplastic transformation of murine NIH3T3 fibroblasts. Whereas simple haploinsufficiency of Sam68 produced by insertion mutagenesis in a single chromosomal allele did not detectably affect cell growth, reduction of Sam68 protein to <25% of the wild type level was associated with anchorage-independent growth, defective contact inhibition, and the ability to form metastatic tumors in nude mice. These properties were reversed by cessation of RHKO inactivation. Our findings, which indicate that the Sam68 protein level can prominently affect cell proliferation, implicate Sam68 function in tumorigenesis. Consistent with these results is evidence that cells undergoing mitosis show a dramatic reduction in the level of Sam68 protein. PMID- 11032832 TI - Dynamin II regulates hormone secretion in neuroendocrine cells. AB - The dynamin family of GTP-binding proteins has been implicated as playing an important role in endocytosis. In Drosophila shibire, mutations of the single dynamin gene cause blockade of endocytosis and neurotransmitter release, manifest as temperature-sensitive neuromuscular paralysis. Mammals express three dynamin genes: the neural specific dynamin I, ubiquitous dynamin II, and predominantly testicular dynamin III. Mutations of dynamin I result in a blockade of synaptic vesicle recycling and receptor-mediated endocytosis. Here, we show that dynamin II plays a key role in controlling constitutive and regulated hormone secretion from mouse pituitary corticotrope (AtT20) cells. Dynamin II is preferentially localized to the Golgi apparatus where it interacts with G-protein betagamma subunit and regulates secretory vesicle release. The presence of dynamin II at the Golgi apparatus and its interaction with the betagamma subunit are mediated by the pleckstrin homology domain of the GTPase. Overexpression of the pleckstrin homology domain, or a dynamin II mutant lacking the C-terminal SH3-binding domain, induces translocation of endogenous dynamin II from the Golgi apparatus to the plasma membrane and transformation of dynamin II from activity in the secretory pathway to receptor-mediated endocytosis. Thus, dynamin II regulates secretory vesicle formation from the Golgi apparatus and hormone release from mammalian neuroendocrine cells. PMID- 11032833 TI - Activation of hepatocyte growth factor (HGF) by endogenous HGF activator is required for metanephric kidney morphogenesis in vitro. AB - The interaction of hepatocyte growth factor (HGF) with c-Met has been implicated in morphogenesis of the kidney, lung, mammary gland, liver, placenta, and limb bud. HGF is secreted as an inactive zymogen and must be cleaved by a serine protease to initiate Met signaling. We show here that a serine protease specific for HGF, HGF activator (HGFA), is expressed and activated by the ureteric bud of the developing kidney in vivo and in vitro. Inhibition of HGFA activity with serine protease inhibitors reduced ureteric bud branching and inhibited glomerulogenesis and nephrogenesis. Activated HGF rescued developing kidneys from the effects of inhibitors. HGFA was localized around the tips of the ureteric bud in developing kidneys, while HGF was expressed diffusely throughout the mesenchyme. These data show that expression of HGF is not sufficient for development, but that its activation is also required. The localization of HGFA to the ureteric bud and the mesenchyme immediately adjacent to it suggests that HGFA creates a gradient of HGF activity in the developing kidney. The creation and shape of gradients of activated HGF by the localized secretion of HGF activators could play an important role in pattern formation by HGF responsive tissues. PMID- 11032834 TI - Cathepsin B-like cysteine proteases confer intestinal cysteine protease activity in Haemonchus contortus. AB - Cathepsin B-like cysteine protease genes (cbls) constitute large multigene families in parasitic and nonparasitic nematodes. Although expressed in the intestine of some nematodes, the biological and biochemical functions of the CBL proteins remain unresolved. Di- and tetra-oligopeptides were used as fluorogenic substrates and irreversible/competitive inhibitors to establish CBL functions in the intestine of the parasitic nematode Haemonchus contortus. Cysteine protease activity was detected against diverse substrates including the cathepsin B/L substrate FR, the caspase 1 substrate YVAD, the cathepsin B substrate RR, but not the CED-3 (caspase 3) substrate DEVD. The pH at which maximum activity was detected varied according to substrate and ranged from pH 5.0 to 7.0. Individual CBLs were affinity isolated using FA and YVAD substrates. pH influenced CBL affinity isolation in a substrate-specific manner that paralleled pH effects on individual substrates. N-terminal sequencing identified two isolated CBLs as H. contortus GCP-7 (33 kDa) and AC-4 (37 kDa). N termini of each began at a position consistent with proregion cleavage and protease activation. Isolation of the GCP 7 band by each peptide was preferentially inhibited when competed with a diazomethane-conjugated inhibitor, Z-FA-CHN(2), demonstrating one functional difference among CBLs and among inhibitors. Substrate-based histological analysis placed CBLs on the intestinal microvilli. Data indicate that CBLs are responsible for cysteine protease activity described from H. contortus intestine. Results also support a role of CBLs in nutrient digestion. PMID- 11032835 TI - A novel superoxide-producing NAD(P)H oxidase in kidney. AB - During phagocytosis, gp91(phox), the catalytic subunit of the phagocyte NADPH oxidase, becomes activated to produce superoxide, a precursor of microbicidal oxidants. Currently increasing evidence suggests that nonphagocytic cells contain similar superoxide-producing oxidases, which are proposed to play crucial roles in various events such as cell proliferation and oxygen sensing for erythropoiesis. Here we describe the cloning of human cDNA that encodes a novel NAD(P)H oxidase, designated NOX4. The NOX4 protein of 578 amino acids exhibits 39% identity to gp91(phox) with special conservation in membrane-spanning regions and binding sites for heme, FAD, and NAD(P)H, indicative of its function as a superoxide-producing NAD(P)H oxidase. The membrane fraction of kidney-derived human embryonic kidney (HEK) 293 cells, expressing NOX4, exhibits NADH- and NADPH dependent superoxide-producing activities, both of which are inhibited by diphenylene iodonium, an agent known to block oxygen sensing, and decreased in cells expressing antisense NOX4 mRNA. The human NOX4 gene, comprising 18 exons, is located on chromosome 11q14.2-q21, and its expression is almost exclusively restricted to adult and fetal kidneys. In human renal cortex, high amounts of the NOX4 protein are present in distal tubular cells, which reside near erythropoietin-producing cells. In addition, overexpression of NOX4 in cultured cells leads to increased superoxide production and decreased rate of growth. The present findings thus suggest that the novel NAD(P)H oxidase NOX4 may serve as an oxygen sensor and/or a regulator of cell growth in kidney. PMID- 11032836 TI - The molecular mechanism of lead inhibition of human porphobilinogen synthase. AB - Human porphobilinogen synthase (PBGS) is a main target in lead poisoning. Human PBGS purifies with eight Zn(II) per homo-octamer; four ZnA have predominantly nonsulfur ligands, and four ZnB have predominantly sulfur ligands. Only four Zn(II) are required for activity. To better elucidate the roles of Zn(II) and Pb(II), we produced human PBGS mutants that are designed to lack either the ZnA or ZnB sites. These proteins, MinusZnA (H131A, C223A) and MinusZnB (C122A, C124A, C132A), each become purified with four Zn(II) per octamer, thus confirming an asymmetry in the human PBGS structure. MinusZnA is fully active, whereas MinusZnB is far less active, verifying an important catalytic role for ZnB and the removed cysteine residues. Kinetic properties of the mutants and wild type proteins are described. Comparison of Pb(II) inhibition of the mutants shows that ligands to both ZnA and ZnB interact with Pb(II). The ZnB ligands preferentially interact with Pb(II). At least one ZnA ligand is responsible for the slow tight binding behavior of Pb(II). The data support a novel model where a high affinity lead site is a hybrid of the ZnA and ZnB sites. We propose that the lone electron pair of Pb(II) precludes Pb(II) to function in PBGS catalysis. PMID- 11032837 TI - Molecular identification and characterization of novel human and mouse concentrative Na+-nucleoside cotransporter proteins (hCNT3 and mCNT3) broadly selective for purine and pyrimidine nucleosides (system cib). AB - The human concentrative (Na(+)-linked) plasma membrane transport proteins hCNT1 and hCNT2 are selective for pyrimidine nucleosides (system cit) and purine nucleosides (system cif), respectively. Both have homologs in other mammalian species and belong to a gene family (CNT) that also includes hfCNT, a newly identified broad specificity pyrimidine and purine Na(+)-nucleoside symporter (system cib) from the ancient marine vertebrate, the Pacific hagfish (Eptatretus stouti). We now report the cDNA cloning and characterization of cib homologs of hfCNT from human mammary gland, differentiated human myeloid HL-60 cells, and mouse liver. The 691- and 703-residue human and mouse proteins, designated hCNT3 and mCNT3, respectively, were 79% identical in amino acid sequence and contained 13 putative transmembrane helices. hCNT3 was 48, 47, and 57% identical to hCNT1, hCNT2, and hfCNT, respectively. When produced in Xenopus oocytes, both proteins exhibited Na(+)-dependent cib-type functional activities. hCNT3 was electrogenic, and a sigmoidal dependence of uridine influx on Na(+) concentration indicated a Na(+):uridine coupling ratio of at least 2:1 for both hCNT3 and mCNT3 (cf 1:1 for hCNT1/2). Phorbol myristate acetate-induced differentiation of HL-60 cells led to the parallel appearance of cib-type activity and hCNT3 mRNA. Tissues containing hCNT3 transcripts included pancreas, bone marrow, trachea, mammary gland, liver, prostate, and regions of intestine, brain, and heart. The hCNT3 gene mapped to chromosome 9q22.2 and included an upstream phorbol myristate acetate response element. PMID- 11032838 TI - The proteasome regulates receptor-mediated endocytosis of interleukin-2. AB - Recent studies have increasingly implicated the proteasome in the regulation of cell surface receptors. In the present study, we investigated the role of the proteasome for ligand-dependent endocytosis and degradation of the interleukin-2 (IL-2)-interleukin-2 receptor (IL-2R) complex. Proteasome inhibitors impaired internalization of IL-2.IL-2R and prevented the lysosomal degradation of this cytokine. Based on time-course studies, proteasome activity is primarily required after initial endocytosis of the IL-2.IL-2R. Proteasome function was also necessary for the lysosomal degradation of IL-2 internalized by IL-2R that were comprised of cytoplasmic tailless beta- or gamma c-subunits, suggesting that the target protein for the proteasome is independent of either the cytoplasmic tail of the IL-2R beta- or gamma c-subunits and their associated signaling components. Therefore, a functional proteasome is required for optimal endocytosis of the IL 2R/ligand complex and is essential for the subsequent lysosomal degradation of IL 2, possibly by regulating trafficking to the lysosome. PMID- 11032839 TI - The structure of the chloroplast F1-ATPase at 3.2 A resolution. AB - The structure of the F(1)-ATPase from spinach chloroplasts was determined to 3.2 A resolution by molecular replacement based on the homologous structure of the bovine mitochondrial enzyme. The crystallized complex contains four different subunits in a stoichiometry of alpha(3)beta(3)gammaepsilon. Subunit delta was removed before crystallization to improve the diffraction of the crystals. The overall structure of the noncatalytic alpha-subunits and the catalytic beta subunits is highly similar to those of the mitochondrial and thermophilic subunits. However, in the crystal structure of the chloroplast enzyme, all alpha- and beta-subunits adopt a closed conformation and appear to contain no bound adenine nucleotides. The superimposed crystallographic symmetry in the space group R32 impaired an exact tracing of the gamma- and epsilon-subunits in the complex. However, clear electron density was present at the core of the alpha(3)beta(3)-subcomplex, which probably represents the C-terminal domain of the gamma-subunit. The structure of the spinach chloroplast F(1) has a potential binding site for the phytotoxin, tentoxin, at the alphabeta-interface near betaAsp(83) and an insertion from betaGly(56)-Asn(60) in the N-terminal beta barrel domain probably increases the thermal stability of the complex. The structure probably represents an inactive latent state of the ATPase, which is unique to chloroplast and cyanobacterial enzymes. PMID- 11032840 TI - Tumor necrosis factor-alpha (TNF) stimulates RANKL-induced osteoclastogenesis via coupling of TNF type 1 receptor and RANK signaling pathways. AB - Tumor necrosis factor-alpha (TNF) and the ligand for receptor activator of NF kappaB (RANKL) are abundant in sites of inflammatory bone erosion. Because these cytokines are potent osteoclastogenic factors and because their signaling pathways are considerably overlapping, we postulated that under pro-inflammatory conditions RANKL and TNF might synergistically orchestrate enhanced osteoclastogenesis via cooperative mechanisms. We found TNF, via TNF type 1 receptor (TNFr1), prompts robust osteoclastogenesis by osteoclast precursors pretreated with RANKL, and deletion of TNFr1 abrogates this response. Enhanced osteoclastogenesis is associated with high expression of otherwise TNF and RANKL induced mediators, including c-Src, TRAF2, TRAF6, and MEKK-1, levels of which were notably reduced in TNFr1 knockouts. Recruitment of TRAFs and MEKK1 leads to activation of downstream pathways, primarily I kappa B/NF-kappa B, ERKs, and cJun/AP-1. Consistent with impaired osteoclastogenesis and reduced expression of TRAFs and MEKK1, we found that phosphorylation and activation of I kappa B, NF kappa B, ERKs, and cJun/AP-1 are severely reduced in RANKL-treated TNFr1-null osteoclast precursors compared with wild type counterparts. Finally, we found that TNF and RANKL synergistically up-regulate RANK expression in wild type precursors, whereas basal and stimulated levels of RANK are significantly lower in TNFr1 knockout cells. Our data suggest that exuberant TNF-induced osteoclastogensis is the result of coupling between RANK and TNFr1 and is dependent upon signals transmitted by the latter receptor. PMID- 11032841 TI - Mechanistic implications of mutations to the active site lysine of porphobilinogen synthase. AB - Porphobilinogen synthase (PBGS) is a homo-octameric protein that catalyzes the complex asymmetric condensation of two molecules of 5-aminolevulinic acid (ALA). The only characterized intermediate in the PBGS-catalyzed reaction is a Schiff base that forms between the first ALA that binds and a conserved lysine, which in Escherichia coli PBGS is Lys-246 and in human PBGS is Lys-252. In this study, E. coli PBGS mutants K246H, K246M, K246W, K246N, and K246G and human PBGS mutant K252G were characterized. Alterations to this lysine result in a disabled but not totally inactive protein suggesting an alternate mechanism in which proximity and orientation are major catalytic devices. (13)C NMR studies of [3,5 (13)C]porphobilinogen bound at the active sites of the E. coli PBGS and the mutants show only minor chemical shift differences, i.e. environmental alterations. Mammalian PBGS is established to have four functional active sites, whereas the crystal structure of E. coli PBGS shows eight spatially distinct and structurally equivalent subunits. Biochemical data for E. coli PBGS have been interpreted to support both four and eight active sites. A unifying hypothesis is that formation of the Schiff base between this lysine and ALA triggers a conformational change that results in asymmetry. Product binding studies with wild-type E. coli PBGS and K246G demonstrate that both bind porphobilinogen at four per octamer although the latter cannot form the Schiff base from substrate. Thus, formation of the lysine to ALA Schiff base is not required to initiate the asymmetry that results in half-site reactivity. PMID- 11032842 TI - Nuclear translocation of the N-terminal prodomain of interleukin-16. AB - Interleukin-16 (IL-16) is a pleiotropic cytokine that functions as a chemoattractant factor, a modulator of T cell activation, and an inhibitor of human immunodeficiency virus (HIV) replication. These diverse functions are exclusively attributed to the secreted C-terminal peptide of 121 amino acids (mature IL-16), which is cleaved from the precursor protein (pro-IL-16) by caspase-3. Human pro-IL-16 is comprised of 631 amino acids with three PDZ domains, one of which is present in secreted mature IL-16. No cellular localization or biologic functions have been ascribed to the unusually large and highly conserved N-terminal prodomain formed as a result of proteolytic release of the third PDZ domain of pro-IL-16. Here we show that the N-terminal prodomain of pro-IL-16 translocates into the nucleus following cleavage of the C-terminal segment. The nuclear localization signal of pro-IL-16 consists of a classical bipartite nuclear targeting motif. We also show that the nuclear targeting of the IL-16 prodomain induces a G(0)/G(1) arrest in the cell cycle. Taken together, the high degree of conservation of the prodomain among species, the presence of two PDZ motifs, and the nuclear localization and subsequent inhibitory effect on cell cycle progression suggest that pro-IL-16 is cleaved into two functional proteins, a C-terminal-secreted cytokine and an N-terminal product, which affects the cell cycle. PMID- 11032843 TI - A novel synthetic acyclic lipid A-like agonist activates cells via the lipopolysaccharide/toll-like receptor 4 signaling pathway. AB - ER-112022 is a novel acyclic synthetic lipid A analog that contains six symmetrically organized fatty acids on a noncarbohydrate backbone. Chinese hamster ovary (CHO)-K1 fibroblasts and U373 human astrocytoma cells do not respond to lipopolysaccharide (LPS) in the absence of CD14. In contrast, exposure to ER-112022 effectively induced activation of CHO and U373 cells under serum free conditions. Expression of CD14 was not necessary for cells to respond to ER 112022, although the presence of soluble CD14 enhanced the sensitivity of the response. Several lines of evidence suggested that ER-112022 stimulates cells via the LPS signal transduction pathway. First, the diglucosamine-based LPS antagonists E5564 and E5531 blocked ER-112022-induced stimulation of CHO-K1, U373, and RAW264.7 cells. Second, ER-112022 was unable to activate C3H/HeJ mouse peritoneal macrophages, containing a mutation in Toll-like receptor (TLR) 4, as well as HEK293 cells, an epithelial cell line that does not express TLR4. Third, ER-112022 activated NF-kappaB in HEK293 cells transfected with TLR4/MD-2. Finally, tumor necrosis factor release from primary human monocytes exposed to ER 112022 was blocked by TLR4 antibodies but not by TLR2 antibodies. Our results suggest that ER-112022 and the family of lipid A-like LPS antagonists can functionally associate with TLR4 in the absence of CD14. Synthetic molecules like ER-112022 may prove to be valuable tools to characterize elements in the LPS receptor complex, as well as to activate or inhibit the TLR4 signaling pathway for therapeutic purposes. PMID- 11032844 TI - The human tissue inhibitor of metalloproteinases (TIMP)-1 gene contains repressive elements within the promoter and intron 1. AB - Expression of the TIMP-1 (tissue inhibitor of metalloproteinases-1) gene is tightly controlled during embryonic development and in the adult animal. Previous studies have focused on elements within the gene promoter which activate transcription of the gene. Here, we identify two regions of the gene which repress transcription: An element upstream of the basal gene promoter at -1718/ 1458, represses expression of a reporter gene by approximately 50%; addition of the first intron to any promoter-reporter construct also strongly represses gene expression. The TIMP-1 gene has a short first exon which is transcribed but not translated, with the translation start site located in exon 2. Deletion analysis through intron 1 reveals a number of potential regions which might mediate its effect. Protein binding studies and mutational analyses reveal that a repressive element at +684/+748 binds Sp1, Sp3, and an unidentified Ets-related factor to suppress transcription. PMID- 11032845 TI - Nitric oxide-inducible expression of heme oxygenase-1 in human cells. Translation independent stabilization of the mRNA and evidence for direct action of nitric oxide. AB - Expression of heme oxygenase-1 (HO-1) in mammalian cells contributes to resistance to various types of free radical damage. Nitric oxide (NO) induces HO 1 in many cell types, but the specific contribution of transcriptional or post transcriptional effects to this induction have remained unresolved. Here we show that the extent of HO-1 mRNA expression in IMR-90 and HeLa cells depends on the rate of NO delivery, and that the induction occurs more slowly in HeLa than in human fibroblast (IMR-90) cells. We used a specific NO scavenger (2-(4 carboxylphenyl)-4,4,5,5-tetramethylimidazolin-1-oxyl 3-oxide) that completely prevented the inducible expression of HO-1 by NO, pointing to direct signaling action of NO in this induction. By inhibiting transcription during the NO exposure, we have confirmed that NO treatment activates a mechanism that stabilizes HO-1 mRNA. The increase in the HO-1 mRNA half-life in IMR-90 cells was directly correlated with increasing rates of NO release. We also show here that the stabilization of the HO-1 message does not require de novo protein synthesis. Collectively, these results show that stabilization of HO-1 mRNA can be finely tuned to the NO exposure, and that the effect in human fibroblasts is mediated by a pre-existing protein. PMID- 11032846 TI - Mutations in the interglobular domain of aggrecan alter matrix metalloproteinase and aggrecanase cleavage patterns. Evidence that matrix metalloproteinase cleavage interferes with aggrecanase activity. AB - We have expressed G1-G2 mutants with amino acid changes at the DIPEN(341) downward arrow(342)FFGVG and ITEGE(373) downward arrow(374)ARGSV cleavage sites, in order to investigate the relationship between matrix metalloproteinase (MMP) and aggrecanase activities in the interglobular domain (IGD) of aggrecan. The mutation DIPEN(341) to DIGSA(341) partially blocked cleavage by MMP-13 and MMP-8 at the MMP site, while the mutation (342)FFGVG to (342)GTRVG completely blocked cleavage at this site by MMP-1, -2, -3, -7, -8, -9, -13, -14. Each of the MMP cleavage site mutants, including a four-amino acid deletion mutant lacking residues ENFF(343), were efficiently cleaved by aggrecanase, suggesting that the primary sequence at the MMP site had no effect on aggrecanase activity in the IGD. The mutation (374)ARGSV to (374)NVYSV completely blocked cleavage at the aggrecanase site by aggrecanase, MMP-8 and atrolysin C but had no effect on the ability of MMP-8 and MMP-13 to cleave at the Asn(341) downward arrowPhe bond. Susceptibility to atrolysin C cleavage at the MMP site was conferred in the DIGSA(341) mutant but absent in the wild-type, (342)GTRVG, (374)NVYSV, and deletion mutants. To further explore the relationship between MMP and aggrecanase activities, sequential digest experiments were done in which MMP degradation products were subsequently digested with aggrecanase and vice versa. Aggrecanase derived G1 domains with ITEGE(373) C termini were viable substrates for MMPs; however, MMP-derived G2 fragments were resistant to cleavage by aggrecanase. A 10 mer peptide FVDIPENFFG, which is a substrate analogue for the MMP cleavage site, inhibited aggrecanase cleavage at the Glu(373) downward arrowAla bond. This study demonstrates that MMPs and aggrecanase have unique substrate recognition in the IGD of aggrecan and suggests that sequences at the C terminus of the DIPEN(341) G1 domain may be important for regulating aggrecanase cleavage. PMID- 11032847 TI - Comparison of the two murine terminal deoxynucleo- tidyltransferase isoforms. A 20-amino acid insertion in the highly conserved carboxyl-terminal region modifies the thermosensitivity but not the catalytic activity PMID- 11032849 TI - Series introduction: emerging clinical applications of nucleic acids PMID- 11032850 TI - Escort aptamers: a delivery service for diagnosis and therapy. PMID- 11032851 TI - Developing aptamers into therapeutics. PMID- 11032852 TI - Oral tolerance, an active immunologic process mediated by multiple mechanisms. PMID- 11032853 TI - A new role of lipid receptors in vascular and cardiac morphogenesis. PMID- 11032854 TI - Osteocyte and osteoblast apoptosis and excessive bone deposition accompany failure of collagenase cleavage of collagen. AB - Mice carrying a targeted mutation (r) in Col1a1, encoding a collagenase-resistant form of type I collagen, have altered skeletal remodeling. In hematoxylin and eosin-stained paraffin sections, we detect empty lacunae in osteocytes in calvariae from Col1a1(r/r) mice at age 2 weeks, increasing through age 10-12 months. Empty lacunae appear to result from osteocyte apoptosis, since staining of osteocytes/periosteal osteoblasts with terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling is increased in Col1a1(r/r) relative to wild-type bones. Osteocyte perilacunar matrices stained with Ab that recognizes collagenase collagen alpha1(I) chain cleavage ends in wild-type but not Col1a1(r/r) calvariae. Increased calvarial periosteal and tibial/femoral endosteal bone deposition was found in Col1a1(r/r) mice from ages 3-12 months. Calcein labeling of calvarial surfaces was increased in Col1a1(r/r) relative to wild-type mice. Daily injections of synthetic parathyroid hormone for 30 days increased calcein surface labeling in wild-type but caused no further increase in the already high calcein staining of Col1a1(r/r) bones. Thus, failure of collagenase cleavage of type I collagen in Col1a1(r/r) mice is associated with osteocyte/osteoblast death but increases bone deposition in a manner that mimics the parathyroid hormone induced bone surface activation seen in wild-type mice. PMID- 11032855 TI - Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation. AB - Sphingolipid signaling pathways have been implicated in many critical cellular events. Sphingosine-1-phosphate (SPP), a sphingolipid metabolite found in high concentrations in platelets and blood, stimulates members of the endothelial differentiation gene (Edg) family of G protein-coupled receptors and triggers diverse effects, including cell growth, survival, migration, and morphogenesis. To determine the in vivo functions of the SPP/Edg signaling pathway, we disrupted the Edg1 gene in mice. Edg1(-/-) mice exhibited embryonic hemorrhage leading to intrauterine death between E12.5 and E14.5. Vasculogenesis and angiogenesis appeared normal in the mutant embryos. However, vascular maturation was incomplete due to a deficiency of vascular smooth muscle cells/pericytes. We also show that Edg-1 mediates an SPP-induced migration response that is defective in mutant cells due to an inability to activate the small GTPase, Rac. Our data reveal Edg-1 to be the first G protein-coupled receptor required for blood vessel formation and show that sphingolipid signaling is essential during mammalian development. PMID- 11032856 TI - Pax3 is required for enteric ganglia formation and functions with Sox10 to modulate expression of c-ret. AB - Hirschsprung disease and Waardenburg syndrome are human genetic diseases characterized by distinct neural crest defects. Patients with Hirschsprung disease suffer from gastrointestinal motility disorders, whereas Waardenburg syndrome consists of defective melanocyte function, deafness, and craniofacial abnormalities. Mutations responsible for Hirschsprung disease and Waardenburg syndrome have been identified, and some patients have been described with characteristics of both disorders. Here, we demonstrate that PAX3, which is often mutated in Waardenburg syndrome, is required for normal enteric ganglia formation. Pax3 can bind to and activate expression of the c-RET gene, which is often mutated in Hirschsprung disease. Pax3 functions with Sox10 to activate transcription of c-RET, and SOX10 mutations result in Waardenburg-Hirschsprung syndrome. Thus, Pax3, Sox10, and c-Ret are components of a neural crest development pathway, and interruption of this pathway at various stages results in neural crest-related human genetic syndromes. PMID- 11032857 TI - Dehydration activates an NF-kappaB-driven, COX2-dependent survival mechanism in renal medullary interstitial cells. AB - Renal prostaglandin (PG) synthesis is mediated by cyclooxygenase-1 and -2 (COX1 and COX2). After dehydration, the maintenance of normal renal function becomes particularly dependent upon PG synthesis. The present studies were designed to examine the potential link between medullary COX1 and COX2 expression in hypertonic stress. In response to water deprivation, COX2, but not COX1, mRNA levels increase significantly in the renal medulla, specifically in renal medullary interstitial cells (RMICs). Water deprivation also increases renal NF kappaB-driven reporter expression in transgenic mice. NF-kappaB activity and COX2 expression could be induced in cultured RMICs with hypertonic sodium chloride and mannitol, but not urea. RMIC COX2 expression was also induced by driving NF kappaB activation with a constitutively active IkappaB kinase alpha (IKKalpha). Conversely, introduction of a dominant-negative IkappaB mutant reduced COX2 expression after hypertonicity or IKKalpha induction. RMICs failed to survive hypertonicity when COX2 was downregulated using a COX2-selective antisense or blocked with the selective nonsteroidal anti-inflammatory drug (NSAID) SC58236, reagents that did not affect cell survival in isotonic media. In rabbits treated with SC58236, water deprivation induced apoptosis of medullary interstitial cells in the renal papilla. These results demonstrate that water deprivation and hypertonicity activate NF-kappaB. The consequent increase in COX2 expression favors RMIC survival in hypertonic conditions. Inhibition of RMIC COX2 could contribute to NSAID-induced papillary injury. PMID- 11032858 TI - Cytosolic phospholipase A(2) regulates golgi structure and modulates intracellular trafficking of membrane proteins. AB - The Golgi complex and the trans-Golgi network are critical cellular organelles involved in the endocytic and biosynthetic pathways of protein trafficking. Lipids have been implicated in the regulation of membrane-protein trafficking, vesicular fusion, and targeting. We have explored the role of cytosolic group IV phospholipase A(2) (cPLA(2)) in membrane-protein trafficking in kidney epithelial cells. Adenoviral expression of cPLA(2) in LLC-PK(1) kidney epithelial cells prevents constitutive trafficking to the plasma membrane of an aquaporin 2-green fluorescent protein chimera, with retention of the protein in the rough endoplasmic reticulum. Plasma membrane Na(+)-K(+)-ATPase alpha-subunit localization is markedly reduced in cells expressing cPLA(2), whereas the trafficking of a Cl(-)/HCO(3)(-) anion exchanger to the plasma membrane is not altered in these cells. Expression of cPLA(2) results in dispersion of giantin and beta-COP from their normal, condensed Golgi localization, and in marked disruption of the Golgi cisternae. cPLA(2) is present in Golgi fractions from noninfected LLC-PK(1) cells and rat kidney cortex. The distribution of tubulin and actin was not altered by cPLA(2), indicating that the microtubule and actin cytoskeleton remain intact. Total cellular protein synthesis is unaffected by the increase in cPLA(2) activity. Thus cPLA(2) plays an important role in determining Golgi architecture and selective control of constitutive membrane-protein trafficking in renal epithelial cells. PMID- 11032859 TI - CD44 as a receptor for colonization of the pharynx by group A Streptococcus. AB - The pharynx is the primary reservoir for strains of group A STREPTOCOCCUS: (GAS) associated both with pharyngitis (streptococcal sore throat) and with invasive or "flesh-eating" soft tissue infections. We now report that CD44, a hyaluronic acid binding protein that mediates human cell-cell- and cell-extracellular matrix binding interactions, functions as a receptor for GAS colonization of the pharynx in vivo. We found that attachment of GAS to murine epithelial keratinocytes was mediated by binding of the GAS hyaluronic acid capsular polysaccharide to CD44. In studies of transgenic mice with a selective defect in epithelial expression of CD44, GAS adherence to CD44-deficient keratinocytes in vitro was reduced compared with adherence to keratinocytes expressing normal levels of CD44. After intranasal inoculation, GAS colonized the oropharynx of wild-type mice but failed to colonize transgenic mice deficient in CD44 expression. GAS colonization of wild-type mice could be blocked by coadministration of mAb to CD44 or by pretreatment of the animals with exogenous hyaluronic acid. These results provide evidence that CD44 serves as a receptor for GAS colonization of the pharynx and support the potential efficacy of disrupting the interaction between the GAS hyaluronic acid capsule and CD44 as a novel approach to preventing pharyngeal infection. PMID- 11032860 TI - CD4 T cell-mediated cardiac allograft rejection requires donor but not host MHC class II. AB - Numerous studies indicate that CD4 T cells are required for acute cardiac allograft rejection. However, the precise role for CD4 T cells in this response has remained ambiguous owing to the multipotential properties of this T-cell subpopulation. In the current study, we demonstrate the capacity of CD4 T cells to serve as direct effector cells of cardiac allograft rejection. We show that CD4 T cells are both necessary and sufficient for acute graft rejection, as indicated by adoptive transfer experiments in immune-deficient SCID and rag1(-/-) recipients. We have analyzed the contribution of direct (donor MHC class II restricted) and indirect (host MHC class II restricted) antigen recognition in CD4-mediated rejection. Acute CD4 T cell-mediated rejection required MHC class II expression by the allograft, indicating the importance of direct graft recognition. In contrast, reciprocal experiments indicate that CD4 T cells can acutely reject allogeneic cardiac allografts established in rag1(-/-) hosts that were also MHC class II deficient. This latter result indicates that indirect presentation of donor antigens by host MHC class II is not required for acute CD4 mediated rejection. Taken together, these results indicate that CD4 T cells can serve as effector cells for primary acute cardiac allograft rejection, predominantly via direct donor antigen recognition and independent of indirect reactivity. PMID- 11032861 TI - Akt1/PKB upregulation leads to vascular smooth muscle cell hypertrophy and polyploidization. AB - Vascular smooth muscle cells (VSMCs) at capacitance arteries of hypertensive individuals and animals undergo marked age- and blood pressure-dependent polyploidization and hypertrophy. We show here that VSMCs at capacitance arteries of rat models of hypertension display high levels of Akt1/PKB protein and activity. Gene transfer of Akt1 to VSMCs isolated from a normotensive rat strain was sufficient to abrogate the activity of the mitotic spindle cell-cycle checkpoint, promoting polyploidization and hypertrophy. Furthermore, the hypertrophic agent angiotensin II induced VSMC polyploidization in an Akt1 dependent manner. These results demonstrate that Akt1 regulates ploidy levels in VSMCs and contributes to vascular smooth muscle polyploidization and hypertrophy during hypertension. PMID- 11032862 TI - Cholecystokinin-B/gastrin receptors enhance wound healing in the rat gastric mucosa. AB - Although physiological functions of the CCK-B/gastrin receptor are well explored, little is known about its role during healing. Here, we evaluated the role of this receptor in the rat oxyntic mucosa following the introduction of a cryoulcer. In this model, we located and quantified CCK-B/gastrin receptors by reverse transcriptase PCR and receptor autoradiography. Rats with cryoulcers were treated with placebo, omeprazole, the CCK-B/gastrin receptor antagonist YF-476, omeprazole plus YF-476, gastrin-17, and gastrin 17 plus YF-476. During wound healing, CCK-B/gastrin receptors were specifically expressed and localized to the regenerative mucosal ulcer margin. This high expression was limited in time, and the pattern of expression of CCK-B/gastrin receptors correlated closely with the proliferative activity of the regenerative mucosa. Functionally, omeprazole and gastrin-17 caused profound hypergastrinemia, increased cell proliferation in the mucosal ulcer margin and accelerated the late ulcer healing phase. These effects were completely reversed by cotherapy with YF-476. These in vivo and vitro data suggest that CCK-B/gastrin receptors in regenerative rat gastric oxyntic mucosa enhance trophic effects during wound healing. PMID- 11032863 TI - T-cell activation and receptor downmodulation precede deletion induced by mucosally administered antigen. AB - The fate of antigen-specific T cells was characterized in myelin basic protein (MBP) T-cell receptor (TCR) transgenic (Tg) mice after oral administration of MBP. Peripheral Th cells are immediately activated in vivo, as indicated by upregulation of CD69 and increased cytokine responses (Th1 and Th2). Concurrently, surface TCR expression diminishes and internal TCR levels increase. When challenged for experimental autoimmune encephalomyelitis during TCR downmodulation, Tg mice are protected from disease. To characterize Th cells at later times after antigen feeding, it was necessary to prevent thymic release of naive Tg cells. Therefore, adult Tg mice were thymectomized before treatment. TCR expression returns in thymectomized Tg mice 3 days after MBP feeding and then ultimately declines in conjunction with MBP-specific proliferation and cytokine responses (Th1-type and Th2-type). The decline correlates with an increase in apoptosis. Collectively, these results demonstrate that a high dose of fed antigen induces early T-cell activation and TCR downmodulation, followed by an intermediate stage of anergy and subsequent deletion. PMID- 11032865 TI - Chronic immune activation associated with intestinal helminth infections results in impaired signal transduction and anergy. AB - Helminthic parasites cause widespread, persistent infections in humans. The immigration of Ethiopians to Israel (a group denoted here by "Eth."), many of them infested with helminths and in a chronic immune-activation state, enabled us to investigate the effects of such immune activation on immune responses. We studied the immune profile and immune functions of 190 Eth. and Israeli non-Eth. (Isr.) highly, partially, or non-immune-activated individuals. Immune cells from highly immune-activated individuals were defective in several signaling responses, all of which were restored gradually following anti-helminthic treatment. These cells showed poor transmembrane signaling, as seen by the phosphorylation of various tyrosine kinases and of the MAPK kinases, ERK1/2 and p38; deficient degradation of phosphorylated IkappaBalpha; increased expression of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), which appears to block proliferative responses in these cells; decreased beta-chemokine secretion by CD8(+) cells after stimulation; and reduced proliferation to recall antigen stimulation. Highly immune-activated individuals also showed decreased delayed type skin hypersensitivity responses to recall antigen before deworming. These findings support the notion that chronic helminthic infections cause persistent immune activation that results in hyporesponsiveness and anergy. Such impaired immune functions may diminish the capacity of these individuals to cope with infections and to generate cellular protective immunity after vaccination. PMID- 11032866 TI - The single-stranded DNA- and RNA-binding proteins pur alpha and pur beta link BC1 RNA to microtubules through binding to the dendrite-targeting RNA motifs. AB - Neural BC1 RNA is distributed in neuronal dendrites as RNA-protein complexes (BC1 RNPs) containing Translin. In this study, we demonstrated that the single stranded DNA- and RNA-binding protein pur alpha and its isoform, pur beta, which have been implicated in control of DNA replication and transcription, linked BC1 RNA to microtubules (MTs). The binding site was within the 5' proximal region of BC1 RNA containing putative dendrite-targeting RNA motifs rich in G and U residues, suggesting that in the cytoplasm of neurons, these nuclear factors are involved in the BC1 RNA transport along dendritic MTs. The pur proteins were not components of BC1 RNP but appeared to associate with MTs in brain cells. Therefore, it is suggested that they may transiently interact with the RNP during transport. In this respect, the interaction of pur proteins with BC1 RNA could be regulated by the Translin present within the RNP, because the binding mode of these two classes of proteins (pur proteins and Translin) to the dendrite targeting RNA motifs was mutually exclusive. As the motifs are well conserved in microtubule-associated protein 2a/b mRNA as well, the pur proteins may also play a role(s) in the dendritic transport of a subset of mRNAs. PMID- 11032864 TI - Differential coreceptor expression allows for independent evolution of non syncytium-inducing and syncytium-inducing HIV-1. AB - We demonstrated previously that CD45RA(+) CD4(+) T cells are infected primarily by syncytium-inducing (SI) HIV-1 variants, whereas CD45RO(+) CD4(+) T cells harbor both non-SI (NSI) and SI HIV-1 variants. Here, we studied evolution of tropism for CD45RA(+) and CD45RO(+) CD4(+) cells, coreceptor usage, and molecular phylogeny of coexisting NSI and SI HIV-1 clones that were isolated from four patients in the period spanning SI conversion. NSI variants were CCR5-restricted and could be isolated throughout infection from CD45RO(+) CD4(+) cells. SI variants seemed to evolve in CD45RO(+) CD4(+) cells, but, in time, SI HIV-1 infection of CD45RA(+) CD4(+) cells equaled infection of CD45RO(+) CD4(+) cells. In parallel with this shift, SI HIV-1 variants first used both coreceptors CCR5 and CXCR4, but eventually lost the ability to use CCR5. Phylogenetically, NSI and SI HIV-1 populations diverged over time. We observed a differential expression of HIV-1 coreceptors within CD45RA(+) and CD45RO(+) cells, which allowed us to isolate virus from purified CCR5(+) CXCR4(-) and CCR5(-) CXCR4(+) CD4(+) cells. The CCR5(+) subset was exclusively infected by CCR5-dependent HIV-1 clones, whereas SI clones were preferentially isolated from the CXCR4(+) subset. The differential expression of HIV-1 coreceptors provides distinct cellular niches for NSI and SI HIV-1, contributing to their coexistence and independent evolutionary pathways. PMID- 11032867 TI - Novel CNS glutamate receptor subunit genes of Drosophila melanogaster. AB - We report the identification and characterization of two genes from Drosophila melanogaster that encode novel ionotropic glutamate receptor proteins, named DGluR-IB and DNMDAR-II, and that are located on chromosome 3L, region 67AB, and the X chromosome, position 2B, respectively. The DGluR-IB full-length cDNA was isolated from Drosophila embryonic and head libraries. The encoded protein of 1,095 amino acids displays high sequence identity (73%) to DGluR-IA. The DNMDAR II gene was identified by sequence-homology searches in databases. The deduced protein shows moderate sequence identity (29-31%) to the mouse NMDAR2A-D receptor subunits. Whole-mount in situ hybridization on embryos revealed DGluR-IB and DNMDAR-II transcripts in the CNS. Immunofluorescence analysis of the adult fly brain indicates that the DGluR-IB protein is expressed in neurons implicated in the regulation of the circadian clock. PMID- 11032868 TI - Characterization of GAR-2, a novel G protein-linked acetylcholine receptor from Caenorhabditis elegans. AB - We have previously identified two G protein-linked acetylcholine receptors (GARs), GAR-1 and GAR-3, in the nematode Caenorhabditis elegans. Whereas GAR-3 is a homologue of muscarinic acetylcholine receptors (mAChRs), GAR-1 is similar to but pharmacologically distinct from mAChRs. In the current work we isolated a new type of GAR using C. elegans genome sequence information. This receptor, named GAR-2, consists of 614 amino acid residues and has seven putative transmembrane domains. Database searches indicate that GAR-2 is most similar to GAR-1 and closely related to GAR-3/mAChRs. The overall amino acid sequence identities to GAR-1 and GAR-3 are approximately 32 and approximately 23%, respectively. When GAR-2 was coexpressed with the G protein-activated inwardly rectifying K(+) (GIRK1) channel in Xenopus oocytes, acetylcholine was able to evoke the GIRK current in a dose-dependent fashion. Oxotremorine, a classical muscarinic agonist, had little effect on the receptor, indicating that GAR-2 is pharmacologically different from mAChRs but rather similar to GAR-1. GAR-2 differs from GAR-1, however, in that it showed virtually no response to muscarinic antagonists such as atropine, scopolamine, and pirenzepine. Expression studies using green fluorescent protein reporter gene fusion revealed that GAR-2 is expressed in a subset of C. elegans neurons, distinct from those expressing GAR-1. Together with our previous reports, this study demonstrates that diverse types of GARs are present in C. elegans. PMID- 11032869 TI - Mice with markedly reduced PACAP (PAC(1)) receptor expression by targeted deletion of the signal peptide. AB - In an attempt to study the pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 (PAC(1)) receptor (PAC(1)R) function in vivo and to produce a mouse model with altered expression of PAC(1)R, we have used gene targeting in embryonic stem cells to disrupt exon 2 of the PAC(1)R gene, which contains the ATG translation start site and the signal peptide. Un-expectedly, active transcription of PAC(1)R mRNA was detected in the mutant mice; however, exon 1 was spliced to exon 3 (skipping exon 2), and (125)I-PACAP27 binding in brain was greatly reduced. PAC(1)R exon 2(-/-) mice were viable, fertile, and morphologically and histologically indistinguishable from their wild-type counterparts. We next examined the ligand binding and cell surface expression of the mutant receptor lacking the signal peptide in transfected COS-7 cells. (125)I PACAP27 binding of the mutant receptor was approximately one-tenth of that in the wild-type receptor. Although the wild-type receptor was expressed abundantly in both the plasma membrane and the cytoplasm around the nucleus, the mutant receptor was expressed in the plasma membrane with a markedly reduced level. Digestion of the membranes with endoglycosidase F greatly reduced the size of the wild-type receptor but only slightly reduced that of the mutant receptor. These results demonstrate that the signal peptide is required for efficient cell surface expression and N-linked glycosylation of the PAC(1)R. However, the mutant receptors still functionally coupled to adenylate cyclase in COS-7 cells, suggesting the presence of sufficient spare receptors such that the mutant receptors are capable of activating the second messenger system. We suggest that the mutant mice with markedly reduced PAC(1)R expression can serve as a useful animal model or cell culture system for further studies in PAC(1)R function. PMID- 11032870 TI - Analysis of three Ptx2 splice variants on transcriptional activity and differential expression pattern in the brain. AB - Three different transcripts of the homeodomain gene termed pituitary homeobox (Ptx) 2 (Pitx2/Brx/Rieg/Solurshin/Arp) were cloned from different species encoding proteins belonging to the paired-like family of homeodomain proteins. Ptx2a (324 amino acids), Ptx2b (271 amino acids), and Ptx2c (318 amino acids) share the C terminus, including the homeodomain, and have different N termini. Here we report the comparative analysis of all three different Ptx2 splice variants for their transcriptional activity and their expression pattern in the adult rat brain. Ptx2 is able to trans-activate via different model promoters in different cell lines. A mild difference in trans-activating potential is observed among the splice variants, but the underlying mechanism is at present unknown. It is surprising that all Ptx2 transcripts displayed an identical expression pattern in the brain. This markedly restricted pattern is limited to the following brain areas: the anterior and intermediate lobes of the pituitary gland, the subthalamic nucleus, the posterior hypothalamic nucleus, the mammillary bodies, the red nucleus, and the deep gray layer of the superior colliculus. The data presented suggest that all variants of Ptx2 are involved in the development and regulation of distinct neuronal cell groups and the pituitary gland. PMID- 11032871 TI - Induction of adrenomedullin during hypoxia in cultured human glioblastoma cells. AB - Adrenomedullin is a potent vasodilator peptide originally isolated from pheochromocytoma. Adrenomedullin is produced by various types of cells including neurons and astrocytes. To explore possible pathophysiological roles of adrenomedullin in hypoxic brain, we studied the effects of hypoxia on the expression of adrenomedullin in T98G human glioblastoma cells by radioimmunoassay and northern blot analysis. Expression levels of adrenomedullin mRNA and immunoreactive adrenomedullin levels in the culture medium were increased by hypoxia about six- and about threefold, respectively. Treatment with cobalt chloride increased expression levels of adrenomedullin mRNA about threefold and immunoreactive adrenomedullin levels in the culture medium about threefold in T98G cells. Using actinomycin D, we showed that hypoxia did not cause the stabilization of the adrenomedullin mRNA, suggesting that the increased adrenomedullin mRNA levels in response to hypoxia are caused mainly by increased transcription. Treatment with cycloheximide caused increases in adrenomedullin mRNA levels in both normoxic and hypoxic states, raising the possibility that some protein(s) may act as a suppressor of adrenomedullin gene expression in T98G cells. These findings indicate that adrenomedullin is highly induced during hypoxia in T98G glioblastoma cells and suggest that increased expression of adrenomedullin during hypoxia may be important in the defense against hypoxia or ischemia in the brain. PMID- 11032872 TI - Messenger RNAs located in myelin sheath assembly sites. AB - The targeting of mRNAs to specific subcellular locations is believed to facilitate the rapid and selective incorporation of their protein products into complexes that may include membrane organelles. In oligodendrocytes, mRNAs that encode myelin basic protein (MBP) and select myelin-associated oligodendrocytic basic proteins (MOBPs) locate in myelin sheath assembly sites (MSAS). To identify additional mRNAs located in MSAS, we used a combination of subcellular fractionation and suppression subtractive hybridization. More than 50% of the 1,080 cDNAs that were analyzed were derived from MBP or MOBP mRNAs, confirming that the method selected mRNAs enriched in MSAS. Of 90 other cDNAs identified, most represent one or more mRNAs enriched in rat brain myelin. Five cDNAs, which encode known proteins, were characterized for mRNA size(s), enrichment in myelin, and tissue and developmental expression patterns. Two of these, peptidylarginine deiminase and ferritin heavy chain, have recognized roles in myelination. The corresponding mRNAs were of different sizes than the previously identified mRNA, and they had tissue and development expression patterns that were indistinguishable from those of MBP mRNA. Three other cDNAs recognize mRNAs whose proteins (SH3p13, KIF1A, and dynein light intermediate chain) are involved in membrane biogenesis. Although enriched in myelin, the tissue and developmental distribution patterns of these mRNAs differed from those of MBP mRNA. Six other cDNAs, which did not share significant sequence homology to known mRNAs, were also examined. The corresponding mRNAs were highly enriched in myelin, and four had tissue and developmental distribution patterns indistinguishable from those of MBP mRNA. These studies demonstrate that MSAS contain a diverse population of mRNAs, whose locally synthesized proteins are placed to contribute to myelin sheath assembly and maintenance. Characterization of these mRNAs and proteins will help provide a comprehensive picture of myelin sheath assembly. PMID- 11032873 TI - Intracellular pH on translocation of protein kinase C isozymes in rat pinealocytes. AB - In rat pinealocytes, cytoplasmic alkalization causes protein kinase C (PKC) translocation, but the isozyme involved is not known. In this study, we investigated the effect of cytoplasmic alkalization on membrane-associated PKCalpha, delta, epsilon, and zeta, four isozymes present in the rat pineal gland. Treatment with NH(4)Cl, which had no effect on PKCzeta, caused a sustained increase in membrane-associated PKCalpha, delta, and epsilon that lasted for at least 60 min. The effect of NH(4)Cl on PKCalpha, delta, and epsilon was reduced by sodium propionate, an agent that counteracts the effect of NH(4)Cl on intracellular pH. Both sodium propionate and 5-(N,N-hexamethylene)amiloride (HMA), two treatments that abolished the effect of norepinephrine on cytoplasmic alkalization, also reduced norepinephrine-mediated increases in membrane associated PKCalpha, delta, and epsilon. In contrast, these two treatments did not have an effect on the increase in membrane-associated PKC isozymes caused by 4beta-phorbol 12-myristate 13-acetate (PMA), an active phorbol ester, even though HMA was effective in abolishing PMA-mediated increases in intracellular pH. These results, apart from demonstrating that cytoplasmic alkalization by itself can cause translocation of PKCalpha, delta, and epsilon in rat pinealocytes, also indicate that the norepinephrine-stimulated cytoplasmic alkalization plays an important role in transducing signals from the adrenergic receptor to selective PKC isozymes. However, PKC translocation stimulated directly by PMA does not appear to be sensitive to changes in intracellular pH. PMID- 11032874 TI - Degradation of the type I inositol 1,4,5-trisphosphate receptor by caspase-3 in SH-SY5Y neuroblastoma cells undergoing apoptosis. AB - The type I inositol 1,4,5-trisphosphate (IP(3)) receptor is selectively down regulated in several neurodegenerative diseases, including Alzheimer's disease, Huntington's chorea, and ischemia, all conditions in which apoptotic neuronal loss occurs. In the present study, we used a neuronal cell line, human neuroblastoma SH-SY5Y cells, to investigate whether the levels of IP(3) receptor are changed during apoptosis in these cells. Following induction of apoptosis by staurosporine, the immunoreactivity of the type I IP(3) receptor in microsome preparations from SH-SY5Y cells was reduced within 2 h, with a further reduction during subsequent hours. Immunoblot analyses, using antibodies to poly(ADP ribose) polymerase and spectrin breakdown products, revealed proteolysis of these caspase-3 substrates within 3 h, confirming that IP(3) receptor cleavage is an early consequence of apoptosis. In vitro incubation of SH-SY5Y microsomes or immunopurified IP(3) receptor from rat cerebellum with recombinant caspase-3 led to generation of immunoreactive breakdown products similar to those observed in intact cells, suggesting that the type I IP(3) receptor is a potential substrate for caspase-3. Preincubation of the neuroblastoma cells with the caspase-3 inhibitor Z-Asp-Glu-Val-Asp-fluoromethyl ketone prevented IP(3) receptor degradation. These results show that the type I IP(3) receptor is a substrate for caspase-3 in neuronal cells and indicate that apoptotic down-regulation of IP(3) receptor levels may contribute to the pathology of neurodegenerative conditions. PMID- 11032875 TI - Nerve tissue-specific (GLUD2) and housekeeping (GLUD1) human glutamate dehydrogenases are regulated by distinct allosteric mechanisms: implications for biologic function. AB - Human glutamate dehydrogenase (GDH), an enzyme central to the metabolism of glutamate, is known to exist in housekeeping and nerve tissue-specific isoforms encoded by the GLUD1 and GLUD2 genes, respectively. As there is evidence that GDH function in vivo is regulated, and that regulatory mutations of human GDH are associated with metabolic abnormalities, we sought here to characterize further the functional properties of the two human isoenzymes. Each was obtained in recombinant form by expressing the corresponding cDNAs in Sf9 cells and studied with respect to its regulation by endogenous allosteric effectors, such as purine nucleotides and branched chain amino acids. Results showed that L-leucine, at 1.0 mM:, enhanced the activity of the nerve tissue-specific (GLUD2-derived) enzyme by approximately 1,600% and that of the GLUD1-derived GDH by approximately 75%. Concentrations of L-leucine similar to those present in human tissues ( approximately 0.1 mM:) had little effect on either isoenzyme. However, the presence of ADP (10-50 microM:) sensitized the two isoenzymes to L-leucine, permitting substantial enzyme activation at physiologically relevant concentrations of this amino acid. Nonactivated GLUD1 GDH was markedly inhibited by GTP (IC(50) = 0.20 microM:), whereas nonactivated GLUD2 GDH was totally insensitive to this compound (IC(50) > 5,000 microM:). In contrast, GLUD2 GDH activated by ADP and/or L-leucine was amenable to this inhibition, although at substantially higher GTP concentrations than the GLUD1 enzyme. ADP and L-leucine, acting synergistically, modified the cooperativity curves of the two isoenzymes. Kinetic studies revealed significant differences in the K:(m) values obtained for alpha-ketoglutarate and glutamate for the GLUD1- and the GLUD2-derived GDH, with the allosteric activators differentially altering these values. Hence, the activity of the two human GDH is regulated by distinct allosteric mechanisms, and these findings may have implications for the biologic functions of these isoenzymes. PMID- 11032876 TI - Cyclic AMP-induced neuronal differentiation via activation of p38 mitogen activated protein kinase. AB - The p38 mitogen-activated protein kinase (MAPK) pathway mediates cellular responses to inflammatory cytokines and environmental stress, but recent studies have indicated that p38 MAPK may be involved in a more widespread set of cellular functions. Here we show that activation of the cyclic AMP (cAMP) pathway induces a rapid, dose-dependent phosphorylation and activation of p38 MAPK and that combined stimulation with forskolin and growth factors results in additive stimulation of p38 MAPK. Forskolin-stimulated neurite out-growth in rat pheochromocytoma PC12 cells was inhibited by the p38 MAPK inhibitor SB203580. With the combination of forskolin and nerve growth factor, neurite outgrowth was additively increased, and this effect was also inhibited by SB203580. Finally, transfection of p38AGF, which exhibits a mutated activation loop, inhibited cAMP mediated neuronal differentiation. The results indicate that p38 MAPK is a downstream target of the cAMP signaling pathway and that p38 MAPK plays a key role in neuronal differentiation induced by cAMP and growth factors by integration of signals from both pathways. PMID- 11032877 TI - Induction of neuronal apoptosis by thiol oxidation: putative role of intracellular zinc release. AB - The membrane-permeant oxidizing agent 2,2'-dithiodipyridine (DTDP) can induce Zn(2+) release from metalloproteins in cell-free systems. Here, we report that brief exposure to DTDP triggers apoptotic cell death in cultured neurons, detected by the presence of both DNA laddering and asymmetric chromatin formation. Neuronal death was blocked by increased extracellular potassium levels, by tetraethylammonium, and by the broad-spectrum cysteine protease inhibitor butoxy-carbonyl-aspartate-fluoromethylketone. N,N,N', N'-Tetrakis-(2 pyridylmethyl)ethylenediamine (TPEN) and other cell-permeant metal chelators also effectively blocked DTDP-induced toxicity in neurons. Cell death, however, was not abolished by the NMDA receptor blocker MK-801, by the intracellular calcium release antagonist dantrolene, or by high concentrations of ryanodine. DTDP generated increases in fluorescence signals in cultured neurons loaded with the zinc-selective dye Newport Green. The fluorescence signals following DTDP treatment also increased in fura-2- and magfura-2-loaded neurons. These responses were completely reversed by TPEN, consistent with a DTDP-mediated increase in intracellular free Zn(2+) concentrations. Our studies suggest that under conditions of oxidative stress, Zn(2+) released from intracellular stores may contribute to the initiation of neuronal apoptosis. PMID- 11032878 TI - The cellular prion protein colocalizes with the dystroglycan complex in the brain. AB - The function of PrP(C), the cellular prion protein (PrP), is still unknown. Like other glycophosphatidylinositol-anchored proteins, PrP resides on Triton insoluble, cholesterol-rich membranous microdomains, termed rafts. We have recently shown that the activity and subcellular localization of the neuronal isoform of nitric oxide synthase (nNOS) are impaired in adult PrP(0/0) mice as well as in scrapie-infected mice. In this study, we sought to determine whether PrP and nNOS are part of the same functional complex and, if so, to identify additional components of such a complex. To this aim, we looked for proteins that coimmunoprecipitated with PrP in the presence of detergents either that completely dissociate rafts, to identify stronger interactions, or that preserve the raft structure, to identify weaker interactions. Using this detergent dependent immunoprecipitation protocol we found that PrP interacts strongly with dystroglycan, a transmembrane protein that is the core of the dystrophin glycoprotein complex (DGC). Additional results suggest that PrP also interacts with additional members of the DGC, including nNOS. PrP coprecipitated only with established presynaptic proteins, consistent with recent findings suggesting that PrP is a presynaptic protein. PMID- 11032879 TI - Characterization of binding sites for chemokines MCP-1 and MIP-1alpha on human brain microvessels. AB - The presence of binding sites for the beta chemokines monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1alpha (MIP-1alpha) has recently been identified on human brain microvessels. We extend these findings in this report to reveal that such sites exemplify characteristics of the recognized major receptors for MCP-1 and MIP-1alpha: CCR2, and CCR1 and CCR5, respectively. Specifically, labeled MCP-1 binding to isolated brain microvessels was inhibited by unlabeled MCP-1 and MCP-3, the latter another CCR2 ligand, but not by MIP 1alpha. Inhibition of labeled MIP-1alpha binding was achieved with unlabeled MIP 1alpha and RANTES, the latter a beta chemokine that binds to both CCR1 and CCR5, but not by MCP-1. Labeled MIP-1alpha binding was also antagonized by unlabeled MCP-3, which is also recognized by CCR1, and MIP-1beta, which is a ligand for CCR5. Labeled MCP-1 and MIP-1alpha were further observed to be internalized within the endothelial cells of brain microvessels, following their binding to the microvascular surface at 37 degrees C. Additionally, exposure of microvessels to unlabeled MCP-1 or MIP-1alpha was accompanied by the initial loss and subsequent recovery of surface binding sites for these chemokines, which occurred on a time scale consistent with ligand-induced endocytosis and recycling. These collective features bear striking similarity to those that characterize interactions of MCP-1 and MIP-1alpha with their receptors on leukocytes and underscore the concept of cognate chemokine receptors on brain microvascular endothelium. PMID- 11032880 TI - Blood-brain barrier is involved in the efflux transport of a neuroactive steroid, dehydroepiandrosterone sulfate, via organic anion transporting polypeptide 2. AB - We have investigated the transport characteristics of dehydroepiandrosterone sulfate (DHEAS), a neuroactive steroid, at the blood-brain barrier (BBB) in a series of functional in vivo and in vitro studies. The apparent BBB efflux rate constant of [(3)H]DHEAS evaluated by the brain efflux index method was 2.68 x 10( 2) min(-1). DHEAS efflux transport was a saturable process with a Michaelis constant (K:(m)) of 32.6 microM: Significant amounts of [(3)H]DHEAS were determined in the jugular venous plasma by HPLC, providing direct evidence that most of the DHEAS is transported in intact form from brain to the circulating blood across the BBB. This efflux transport of [(3)H]DHEAS was significantly inhibited by common rat organic anion-transporting polypeptide (oatp) substrates such as taurocholate, cholate, sulfobromophthalein, and estrone-3-sulfate. Moreover, the apparent efflux clearance of [(3)H]DHEAS across the BBB (118 microl/min-g of brain) was 10.4-fold greater than its influx clearance estimated by the in situ brain perfusion technique (11.4 microl/min-g of brain), suggesting that DHEAS is predominantly transported from the brain to blood across the BBB. In cellular uptake studies using a conditionally immortalized mouse brain capillary endothelial cell line (TM-BBB4), [(3)H]DHEAS uptake by TM-BBB4 cells exhibited a concentration dependence with a K:(m) of 34.4 microM: and was significantly inhibited by the oatp2-specific substrate digoxin. Conversely, [(3)H]digoxin uptake by TM-BBB4 cells was significantly inhibited by DHEAS. Moreover, the net uptake of [(3)H]DHEAS at 30 min was significantly increased under ATP-depleted conditions, suggesting that an energy-dependent efflux process may also be involved in TM-BBB4. RT-PCR and sequence analysis suggest that an oatp2 is expressed in TM-BBB4 cells. In conclusion, DHEAS efflux transport takes place across the BBB, and studies involving in vitro DHEAS uptake and RT-PCR suggest that there is oatp2-mediated DHEAS transport at the BBB. PMID- 11032881 TI - Polyamine uptake in cultured astrocytes: characterization and modulation by protein kinases. AB - The properties and regulation of the polyamine transport system in brain are still poorly understood. The present study shows, for the first time, the existence of a polyamine transport system in cerebellar astrocytes and suggests that polyamine uptake is mediated by a single and saturable high-affinity transport system for putrescine, spermine, and spermidine (K:(m) = 3.2, 1.2, and 1.8 microM:, respectively). Although substitution of NaCl by choline chloride produced a decrease in the putrescine, spermine, and spermidine uptake, it seems that polyamine transport in cerebellar astrocytes is not mediated by an Na(+) cotransport as in the presence of Na(+) and cholinium, polyamine uptake was much lower than when measured in a sucrose-based medium. On the other hand, ouabain, gramicidin (a Na(+) ionophore), and ionomycin (a Ca(2+) ionophore) produced a strong inhibition of polyamine uptake, suggesting that membrane potential could have an important role in the functioning of the astroglial polyamine uptake system. Moreover, protein kinase C inhibition produced an enhancement of polyamine uptake, whereas stimulation of protein kinase C with phorbol esters inhibited polyamine uptake. Alternatively, the tyrosine kinase inhibitor genistein caused a marked reduction in the uptake. No effects on polyamine uptake were observed with inhibitors and activators of cyclic AMP-dependent protein kinase or when Ca(2+)/calmodulin-dependent protein kinase II was inhibited with KN-62. These results suggest that the polyamine uptake system in cerebellar astrocytes could be modulated by protein kinase C and tyrosine kinase activities. PMID- 11032882 TI - Progressive segregation of unmyelinated axons in peripheral nerves, myelin alterations in the CNS, and cyst formation in the kidneys of myelin and lymphocyte protein-overexpressing mice. AB - Myelin and lymphocyte protein (MAL) is a putative tetraspan proteolipid that is highly expressed by Schwann cells and oligodendrocytes as a component of compact myelin. Outside of the nervous system, MAL is found in apical membranes of epithelial cells, mainly in the kidney and stomach. Because MAL is associated with glycosphingolipids, it is thought to be involved in the organization, transport, and maintenance of glycosphingolipid-enriched membrane microdomains. In this report, we describe the generation and analysis of transgenic mice with increased MAL gene dosage. Immunohistochemical analysis revealed that the localization of MAL overexpression in the transgenic animals corresponded closely to the MAL expression pattern observed in wildtype animals, indicating correct spatial regulation of the transgene. Phenotypically, MAL overexpression led to progressive dissociation of unmyelinated axons from bundles in the PNS, a tendency to hypomyelination and aberrant myelin formation in the CNS, and the formation of large cysts in the tubular region of the kidney. Thus, increased expression of MAL appears to be deleterious to membranous structures in the affected tissues, indicating a requirement for tight control of endogenous MAL expression in Schwann cells, oligodendrocytes, and kidney epithelial cells. PMID- 11032883 TI - Transcriptional regulation of 2',3'-cyclic nucleotide 3'-phosphodiesterase gene expression by cyclic AMP in C6 cells. AB - It was recently shown that the two transcripts encoding the isoforms of 2',3' cyclic nucleotide 3'-phosphodiesterase (CNP1 and CNP2) are differentially regulated during the process of oligodendrocyte maturation. In oligodendrocyte precursors, only CNP2 mRNA is present, whereas in differentiating oligodendrocytes, both CNP1 and CNP2 mRNAs are expressed. This pattern of CNP expression is likely due to stage-specific transcriptional regulation of the two CNP promoters during the process of oligodendrocyte differentiation. Here, we report the influence of increased intracellular cyclic AMP (cAMP) levels on the transcription of both CNP1 and CNP2 mRNAs in rat C6 glioma cells. We found that the transcription of CNP1 mRNA was significantly increased in comparison with that of CNP2 mRNA in cells treated with cAMP analogues to elevate intracellular cAMP levels. This up-regulation of CNP1 expression (a) is due to an increase of transcription, (b) requires de novo protein synthesis, and (c) requires the activity of protein kinase A. These results are physiologically significant and support the idea that a cAMP-mediated pathway is part of the molecular mechanisms regulating the expression of CNP1 in oligodendrocytes. The regulation of CNP1 promoter activity by cAMP was then investigated in stably transfected C6 cell lines containing various deletions of the CNP promoter directing the bacterial chloramphenicol acetyltransferase gene. We showed that the sequence between nucleotides -126 and -102 was essential for the cAMP-dependent induction of CNP1 expression. Gel retardation analysis showed that two protein-DNA complexes are formed between this sequence and nuclear factors from C6 cells treated or not treated with cAMP. This suggests that the induction of CNP1 mRNA transcription is not mediated by changes in binding of nuclear factors that interact directly with the -126/-102 sequence. Sequence analysis of this region revealed the presence of a putative activator protein-2 (AP-2) binding site. It is interesting that mutagenesis of this region resulted in a significant reduction in transcriptional responses to cAMP, implying a possible role for the AP-2 factor in the expression of CNP1. In addition, we have shown that putative binding sites for activator protein-4 and nuclear factor-1 adjacent to the AP-2 site are required for efficient induction of CNP1 expression by cAMP. Taken together, our results show that the cAMP-dependent accumulation of CNP1 mRNA appears to depend on the synergistic interaction of several regulatory elements. PMID- 11032884 TI - Impaired mitochondrial function results in increased tissue transglutaminase activity in situ. AB - Tissue transglutaminase (tTG) is a transamidating enzyme that is elevated in Huntington's disease (HD) brain and may be involved in the etiology of the disease. Further, there is evidence of impaired mitochondrial function in HD. Therefore, in this study, we examined the effects of mitochondrial dysfunction on the transamidating activity of tTG. Neuroblastoma SH-SY5Y cells stably overexpressing human tTG or mutated inactive tTG were treated with 3 nitropropionic acid (3-NP), an irreversible inhibitor of succinate dehydrogenase. 3-NP treatment of tTG-expressing cells resulted in a significant increase of TG activity in situ. In vitro measurements demonstrated that 3-NP had no direct effect on tTG activity. However, 3-NP treatment resulted in a significant decrease of the levels of GTP and ATP, two potent inhibitors of the transamidating activity of tTG. No significant changes in the intracellular levels of calcium were observed in 3-NP-treated cells. Treatment with 3-NP in combination with antioxidants significantly reduced the 3-NP-induced increase in in situ TG activity, demonstrating that oxidative stress is a contributing factor to the increase of TG activity. This study demonstrates for the first time that impairment of mitochondrial function significantly increases TG activity in situ, a finding that may have important relevance to the etiology of HD. PMID- 11032885 TI - Thyroid hormone-induced morphological differentiation and maturation of astrocytes are mediated through the beta-adrenergic receptor. AB - The molecular mechanisms associated with thyroid hormone (TH)-induced maturation of astrocytes have been studied using primary cultures. We have previously demonstrated that unlike normal astrocyte cultures, hypothyroid cultures fail to differentiate from flat polygonal cells with epithelioid morphology into mature process-bearing cells with stellate morphology. Addition of TH to the hypothyroid cells reverses the effect, and astrocytes transform into stellate cells. The beta adrenergic receptor (beta-AR) agonist isoproterenol (ISP) has a similar effect, whereas simultaneous addition of the beta-adrenergic antagonist propranolol blocks the differentiation induced by TH or ISP. Addition of TH or ISP to hypothyroid cultures is also associated with a decrease in the level of filamentous cytoskeletal (F(i)) actin and an increase in the level of actin mRNA. Although addition of propranolol inhibited the decline in the level of F(i) actin in the TH- or ISP-supplemented cells as well as the induction of actin mRNA by TH, it partially inhibited the ISP-induced actin mRNA in these cultures. The hormone-induced maturation appears to be selectively regulated through the beta(2)-AR. The overall results indicate that the beta-adrenergic system plays an obligatory role in promoting TH-induced differentiation and maturation of astrocytes and in regulating the hormone-induced expression of actin and its intracellular organization in a way conducive to the morphological differentiation of the cells. PMID- 11032886 TI - Glycosphingolipid composition of a new immortalized human cerebromicrovascular endothelial cell line. AB - Previous studies have demonstrated the involvement of glycosphingolipid (GSL) antigens in the pathogenesis of immune-mediated neurological disorders such as peripheral neuropathies and multiple sclerosis. To study the role of the blood brain barrier (BBB) in these disorders, we used a new human cerebromicrovascular endothelial cell (HCEC) line that has been immortalized through transfection with the plasmid pSV3-neo encoding for the SV40 large T-antigen and the neomycin gene. The immortalized HCEC (SV-HCEC) exhibited accelerated proliferation rates but maintained phenotypic properties of early-passage control cells. Therefore, this human cell line may serve as a useful in vitro model for studying the properties of the human BBB. We first investigated the GSL composition of cultured SV-HCECs. The major gangliosides were GM3 (62% of total gangliosides), GM2 (18%), GM1 (3%), and GD1a (15%). The major neutral GSLs were glucosylceramide (15% of the total neutral glycolipids), lactosylceramide (36%), globotriaosylceramide (3%), and globoside (43%). Trace amounts of paragloboside, lactosaminyl paragloboside, and sulfoglucuronyl paragloboside could also be detected by TLC-immunostaining. These results provide the basis for further investigations of the expression of these cell surface antigens in cultured SV-HCECs on activation with inflammatory cytokines such as interleukin-1beta, tumor necrosis factor-alpha, and interferon gamma, which have been implicated as playing an important role in the pathogenesis of many nervous system disorders. PMID- 11032887 TI - Reversible inactivation of superoxide-sensitive aconitase in Abeta1-42-treated neuronal cell lines. AB - The activity of the superoxide-sensitive enzyme aconitase was monitored to evaluate the generation of superoxide in neuronal cell lines treated with beta amyloid (Abeta) peptide 1-42. Treatment of differentiated and undifferentiated rat PC12 and human neuroblastoma SK-N-SH cells with soluble Abeta1-42 (Abeta derived diffusible ligands) or fibrillar Abeta1-42 caused a 35% reversible inactivation of aconitase, which preceded loss of viability and was correlated with altered cellular function. Aconitase was reactivated upon incubation of cellular extracts with iron and sulfur, suggesting that Abeta causes the release of iron from 4Fe-4S clusters. Abeta neurotoxicity was partially blocked by the iron chelator deferoxamine. These data suggest that increased superoxide generation and the release of iron from 4Fe-4S clusters are early events in Abeta1-42 neurotoxicity. PMID- 11032888 TI - Evidence for a G protein-coupled gamma-hydroxybutyric acid receptor. AB - gamma-Hydroxybutyric acid (GHB) is a naturally occurring metabolite of GABA that has been postulated to exert ubiquitous neuropharmacological effects through GABA(B) receptor (GABA(B)R)-mediated mechanisms. The alternative hypothesis that GHB acts via a GHB-specific, G protein-coupled presynaptic receptor that is different from the GABA(B)R was tested. The effect of GHB on regional and subcellular brain adenylyl cyclase in adult and developing rats was determined and compared with that of the GABA(B)R agonist (-)-baclofen. Also, using guanosine 5'-O:-(3-[(35)S]thiotriphosphate) ([(35)S]GTPgammaS) binding and low K:(m) GTPase activity as markers the effects of GHB and (-)-baclofen on G protein activity in the brain were determined. Neither GHB nor baclofen had an effect on basal cyclic AMP (cAMP) levels. GHB significantly decreased forskolin-stimulated cAMP levels by 40-50% in cortex and hippocampus but not thalamus or cerebellum, whereas (-)-baclofen had an effect throughout the brain. The effect of GHB on adenylyl cyclase was observed in presynaptic and not postsynaptic subcellular tissue preparations, but the effect of baclofen was observed in both subcellular preparations. The GHB-induced alteration in forskolin-induced cAMP formation was blocked by a specific GHB antagonist but not a specific GABA(B)R antagonist. The (-)-baclofen-induced alteration in forskolin-induced cAMP formation was blocked by a specific GABA(B)R antagonist but not a specific GHB antagonist. The negative coupling of GHB to adenylyl cyclase appeared at postnatal day 21, a developmental time point that is concordant with the developmental appearance of [(3)H]GHB binding in cerebral cortex, but the effects of (-)-baclofen were present by postnatal day 14. GHB and baclofen both stimulated [(35)S]GTPgammaS binding and low-K:(m) GTPase activity by 40-50%. The GHB-induced effect was blocked by GHB antagonists but not by GABA(B)R antagonists and was seen only in cortex and hippocampus. The (-)-baclofen-induced effect was blocked by GABA(B)R antagonists but not by GHB antagonists and was observed throughout the brain. These data support the hypothesis that GHB induces a G protein-mediated decrease in adenylyl cyclase via a GHB-specific G protein-coupled presynaptic receptor that is different from the GABA(B)R. PMID- 11032889 TI - Involvement of alpha7 nicotinic acetylcholine receptors in activation of tyrosine hydroxylase and dopamine beta-hydroxylase gene expression in PC12 cells. AB - Nicotine treatment increases intracellular free Ca(2+) concentration [Ca(2+)](i), stimulates catecholamine release, and elevates gene expression for the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DBH). However, the type of nicotinic acetylcholine receptors (nAChRs) mediating these events is unclear. The nAChR receptor antagonists alpha bungarotoxin (alphaBTX) and methyllycaconitine greatly reduced the nicotine triggered initial transient rise in [Ca(2+)](i) and prevented the second prolonged elevation of [Ca(2+)](i), suggesting the involvement of alpha7 nAChRs. Two specific alpha7 nicotinic agonists, 3-(2,4-dimethoxybenzilidene)anabaseine (DMXB) and E, E-3-(cinnamylidene)anabaseine (3-CA), were found to elicit a small, delayed increase in [Ca(2+)](i) with kinetics and magnitude similar to the second elevation observed with nicotine. This increase was inhibited by the inositol trisphosphate receptor antagonist xestospongin C. Exposure to 3-CA or DMXB for 6 or 24 h elevated TH and DBH mRNA levels two- to fourfold over control levels. These agonists were more effective than nicotine alone in increasing TH and DBH gene expression and significantly elevated [Ca(2+)](i) for up to 6 h. The increase in [Ca(2+)](i) or the elevation in TH mRNA by 3-CA was completely inhibited by alphaBTX. This study, for the first time, implicates stimulation of alpha7 nAChRs in the activation of TH and DBH gene expression. PMID- 11032890 TI - Tyrosine phosphorylation of the alpha subunit of transducin and its association with Src in photoreceptor rod outer segments. AB - Recent evidence indicates that tyrosine phosphorylation may play important roles in retinal photoreceptor rod outer segments (ROS). We investigated the tyrosine phosphorylation of endogenous proteins in isolated bovine ROS. Several proteins with apparent molecular masses of 31, 39, 60, 83, 90, 97, 120, 140, and 180 kDa were tyrosine-phosphorylated in ROS incubated with Mg(2+), ATP, and orthovanadate. Several tyrosine kinase inhibitors significantly inhibited tyrosine phosphorylation of these proteins in ROS. The 39- and 60-kDa tyrosine phosphorylated proteins were identified as the alpha subunit of the G protein transducin (Talpha) and the tyrosine kinase Src, respectively. The presence of Src and tyrosine kinase activity in bovine ROS was confirmed by their cofractionation with rhodopsin and Talpha on continuous sucrose gradients. Several tyrosine-phosphorylated proteins, including Src, coimmunoprecipitated with Talpha. The association of Src with Talpha was detected in the absence of tyrosine phosphorylation, but was enhanced with increased tyrosine phosphorylation of ROS. Moreover, tyrosine kinase activity also associated with Talpha was sevenfold higher under tyrosine-phosphorylating conditions. The recovery of transducin by hypotonic GTP extraction from tyrosine-phosphorylated ROS was significantly less than that from nonphosphorylated ROS. We localized the site on Talpha phosphorylated by Src to the amino-terminal half by limited tryptic digests, and further mapped it by ion trap mass spectrometry to Tyr(142) in the helical domain of Talpha. Talpha was also tyrosine-phosphorylated in vivo in rat retina, but this phosphorylation was not affected by light. PMID- 11032891 TI - Interleukin-1beta induces cyclooxygenase-2 and prostaglandin E(2) synthesis in human neuroblastoma cells: involvement of p38 mitogen-activated protein kinase and nuclear factor-kappaB. AB - Prostaglandins (PGs), which are generated by the enzymatic activity of cyclooxygenase (COX)-1 and -2, modulate several functions in the CNS such as the generation of fever, the sleep/wake cycle, and the perception of pain. Moreover, the neuronal induction of COX-2 has been linked to neuroinflammatory aspects of Alzheimer's disease (AD). The regulation of COX expression in neuronal cells is only partly understood and has been mainly linked to synaptic activity. In pathophysiological situations, however, cytokines may be potent stimulators of neuronal COX expression. Here we show that interleukin (IL)-1beta induces COX-2 mRNA and protein synthesis and the release of PGE(2) in the human neuroblastoma cell line SK-N-SH. We further demonstrate that both a free radical scavenger and an inhibitor of p38 mitogen-activated protein kinase (MAPK) reduce IL-1beta induced synthesis of COX-2. IL-1beta induces p38 MAPK phosphorylation and activation of the nuclear factor-kappaB independently from each other. Our data suggest that IL-1beta-induced COX-2 expression in SK-N-SH cells is regulated by different mechanisms, presumably involving mRNA transcription and mRNA stability. The ability of p38 MAPK to augment COX-2 expression in human neuroblastoma cells, as shown here, suggests that p38 MAPK may be involved in neuronal expression of COX-2 in AD. PMID- 11032892 TI - Nitric oxide-sensitive guanylyl cyclase activity inhibition through cyclic GMP dependent dephosphorylation. AB - The soluble form of guanylyl cyclase (sGC) plays a pivotal role in the transduction of inter- and intracellular signals conveyed by nitric oxide. Here, a feedback inhibitory mechanism triggered by cyclic guanosine-3',5'-monophosphate (cGMP)-dependent protein kinase (PKG) activation is described. Preincubation of chromaffin cells with C-type natriuretic peptide, which increased cGMP levels and activated PKG, or with cGMP-permeant analogue (which also activates PKG), in the presence of a broad-spectrum phosphodiesterase inhibitor, resulted in a decrease in subsequent sodium nitroprusside (SNP)-dependent cGMP elevations. This inhibitory effect was mimicked by activating a protein phosphatase and counteracted by the selective PKG inhibitor KT-5823 and by different protein phosphatase inhibitors. Immunoprecipitation of sGC from cells submitted to different treatments followed by immunodetection with antiphosphoserine antibodies (clone 4A9) showed changes in phosphorylation levels of the beta subunit of sGC, and these changes correlated well with differences in SNP elicited cGMP accumulations. Pretreatment of cells with several PKG inhibitors or protein phosphatase inhibitors produced an enhancement of SNP-stimulated cGMP rises without changing the SNP concentration required to produce half-maximal or maximal responses. Taken together, these results indicate that the catalytic activity of sGC is closely coupled to the phosphorylation state of its beta subunit and that the tonic activity of PKG or its stimulation regulates sGC activity through dephosphorylation of the beta subunit. PMID- 11032893 TI - Regulation of NMDA receptor subunits and nitric oxide synthase expression during cocaine withdrawal. AB - The present study characterized the effects of withdrawal from cocaine on the expression of NMDA receptor subunits (NR1, NR2B) and neuronal nitric oxide synthase. FosB induction was measured to confirm that repeated cocaine exposure influenced protein expression, as previously reported. Administration of cocaine followed by 24 h, 72 h, or 14 days of withdrawal resulted in alterations of NR1 and NR2B subunits and neuronal nitric oxide synthase expression as measured by immunohistochemical labeling of rat brain sections. Optical density analyses revealed significant up-regulation of NR1 in the ventral tegmental area at 72 h and 14 days of withdrawal. Structure-specific and withdrawal time-dependent alterations in NR2B expression were also found. After 24 h of withdrawal, cocaine induced decreases in NR2B expression were observed in the nucleus accumbens shell, whereas increases in NR2B expression were found in medial cortical areas. Two weeks of withdrawal from cocaine caused an approximately 50% increase in NR2B subunit expression in regions of the cortex, neostriatum, and nucleus accumbens. In contrast, cocaine-induced up-regulation of neuronal nitric oxide synthase was transient and evident in cortical areas only at 24 h after the last drug injection. The results suggest that region-specific changes in interactions among proteins associated with the NMDA receptor complex may underlie neuronal adaptations following repeated cocaine administration. PMID- 11032894 TI - Some metabotropic glutamate receptor ligands reduce kynurenate synthesis in rats by intracellular inhibition of kynurenine aminotransferase II. AB - Some metabotropic glutamate receptor (mGluR) ligands, such as quisqualate, L-(+) 2-amino-4-phosphonobutyric acid (L-AP4), 4-carboxy-3-hydroxyphenylglycine (4C3HPG), and L-serine-O:-phosphate (L-SOP), reduced the formation of the endogenous excitatory amino acid receptor antagonist kynurenate in brain and liver slices. The use of novel, subtype-selective mGluR agonists and antagonists excluded a role for any known mGluR subtype in this effect. The reduction of kynurenate formation was no longer observed when slices were incubated with the active mGluR ligands in the absence of extracellular Na(+). trans-Pyrrolidine-2,4 dicarboxylate (trans-PDC), a broad-spectrum ligand of Na(+)-dependent glutamate transporters, was also able to reduce kynurenate formation. Quisqualate, 4C3HPG, L-AP4, and L-SOP did not further reduce kynurenate formation in the presence of trans-PDC, suggesting that the two classes of drugs may share the same mechanism of action. Hence, we hypothesized that the active mGluR ligands are transported inside the cell and act intracellularly to reduce kynurenate synthesis. We examined this possibility by assessing the direct effect of mGluR ligands on the activity of kynurenine aminotransferases (KATs) I and II, the enzymes that transaminate kynurenine to kynurenate. In brain tissue homogenates, KAT II (but not KAT I) activity was inhibited by quisqualate, 4C3HPG, L-AP4, L-SOP, and trans PDC. Drugs that were unable to reduce kynurenate formation in tissue slices were inactive. We conclude that some mGluR ligands act intracellularly, inhibiting KAT II activity and therefore reducing kynurenate formation. This effect should be taken into consideration when novel mGluR ligands are developed for the treatment of neurological and psychiatric diseases. PMID- 11032895 TI - Cocaine and methamphetamine differentially affect opioid peptide mRNA expression in the striatum. AB - In general, administration of methamphetamine and cocaine alters preprodynorphin and preproenkephalin mRNA levels in striatum. However, no study has directly compared the effects of these stimulants on opioid peptides in striatum. This study used in situ hybridization to compare directly the effects of cocaine and methamphetamine on preprodynorphin and preproenkephalin mRNAs in distinct striatal regions. Male Sprague-Dawley rats received a single administration of 15 mg/kg methamphetamine or 30 mg/kg cocaine and were killed 30 min or 3 h later. Methamphetamine and cocaine differentially affected preprodynorphin mRNA in striatum after 3 h. Densitometric analysis of film autoradiograms revealed that cocaine, but not methamphetamine, significantly increased preprodynorphin. This effect was seen throughout rostral striatum and dorsally in caudal striatum. However, specific analysis of "patches" in which preprodynorphin expression is high revealed a significantly greater effect of methamphetamine versus cocaine. In contrast, both cocaine and methamphetamine had similar effects on preproenkephalin mRNA, decreasing levels after 30 min in rostral striatum and in the core of nucleus accumbens. These data suggest that methamphetamine and cocaine have distinct postsynaptic consequences on striatal neurons. PMID- 11032896 TI - Coexpression of several types of metabotropic nucleotide receptors in single cerebellar astrocytes. AB - We have examined the expression of mRNA for several P2Y nucleotide receptors by northern blot analysis in purified type 1 cerebellar astrocyte cultures. These results suggest that different P2Y subtypes could be responsible for ATP metabotropic calcium responses in single type 1 astrocytes. To identify these subtypes we have studied the pharmacological profile of ATP calcium responses using fura-2 microfluorimetry. All tested astrocytes responded to ATP and UTP stimulations evoking similar calcium transients. Most astrocytes also responded to 2-methylthioATP and ADP challenges. The agonist potency order was 2 methylthioATP > ADP > ATP = UTP. Cross-desensitization experiments carried out with ATP, UTP, and 2-methylthioATP showed that 2-methylthioATP and UTP interact with different receptors, P2Y(1) and P2Y(2) or P2Y(4). In a subpopulation of type 1 astrocytes, ATP prestimulation did not block UTP responses, and UDP elicited clear intracellular Ca(2+) concentration responses at very low concentrations. 2 MethylthioATP and UTP calcium responses exhibited different sensitivity to pertussis toxin and different inhibition patterns in response to P2 antagonists. The P2Y(1)-specific antagonist N:(6)-methyl-2'-deoxyadenosine 3', 5'-bisphosphate (MRS 2179) specifically blocked the 2-methylthio-ATP responses. We can conclude that all single astrocytes coexpressed at least two types of P2Y metabotropic receptors: P2Y(1) and either P2Y(2) or P2Y(4) receptors. Moreover, 30-40% of astrocytes also coexpressed specific pyrimidine receptors of the P2Y(6) subtype, highly selective for UDP coupled to pertussis-toxin insensitive G protein. PMID- 11032897 TI - Changes in the cannabinoid receptor binding, G protein coupling, and cyclic AMP cascade in the CNS of rats tolerant to and dependent on the synthetic cannabinoid compound CP55,940. AB - Chronic exposure to CP55,940 produced a significant down-regulation of cannabinoid receptors in the striatum, cortex, hippocampus, and cerebellum of rat brain. At 24 h after SR141716-precipitated withdrawal, we observed a tendency to return to basal levels in the striatum and cortex, whereas the specific binding remained lower in the hippocampus and cerebellum. When we surveyed cannabinoid receptor-activated G proteins, in chronic CP55,940-treated rats the guanosine 5' O:-(3-[(35)S]thiotriphosphate) ([(35)S]GTPgammaS) binding assay revealed a decrease of activated G proteins in the striatum, cortex, and hippocampus, whereas no significant changes were seen in the cerebellum. At 24 h after the SR141716-precipitated withdrawal, [(35)S]GTPgammaS binding increased compared with that of rats chronically exposed to CP55,940, attaining the control level except for cerebellum, where we observed a trend to overcome the control amounts. Concerning the cyclic AMP (cAMP) cascade, which represents the major intracellular signaling pathway activated by cannabinoid receptors, in the cerebral areas from rats chronically exposed to CP55,940 we found alteration in neither cAMP levels nor protein kinase A activity. In the brain regions taken from CP55, 940-withdrawn rats, we only observed a significant up-regulation in the cerebellum. Our findings suggest that receptor desensitization and down regulation are strictly involved in the development of cannabinoid tolerance, whereas alterations in the cAMP cascade in the cerebellum could be relevant in the mediation of the motor component of cannabinoid abstinence. PMID- 11032898 TI - Electrically evoked release of [(3)H]noradrenaline from mouse cultured sympathetic neurons: release-modulating heteroreceptors. AB - Cultured neurons from the thoracolumbar sympathetic chain of newborn mice are known to possess release-inhibiting alpha(2)-autoreceptors. The present study was carried out in a search for release-modulating heteroreceptors on these neurons. Primary cultures were preincubated with [(3)H]noradrenaline and then superfused and stimulated by single pulses, trains of 8 pulses at 100 Hz, or trains of 36 pulses at 3 Hz. The cholinergic agonist carbachol reduced the evoked overflow of tritium. Experiments with antagonists indicated that the inhibition was mediated by M(2) muscarinic receptors. The cannabinoid agonist WIN 55,212-2 reduced the evoked overflow of tritium through CB(1) receptors. Prostaglandin E(2), sulprostone, and somatostatin also caused presynaptic inhibition. The inhibitory effects of carbachol, WIN 55,212-2, prostaglandin E(2), and somatostatin were abolished (at the highest concentration of WIN 55, 212-2 almost abolished) by pretreatment of the cultures with pertussis toxin (250 ng/ml). Several drugs, including the beta(2)-adrenoceptor agonist salbutamol, opioid receptor agonists, neuropeptide Y, angiotensin II, and bradykinin, failed to change the evoked overflow of tritium. These results demonstrate a distinct pattern of presynaptic inhibitory heteroreceptors, all coupled to pertussis toxin-sensitive G proteins. The lack of operation of several presynaptic receptors known to exist in adult mice in situ may be due to the age of the (newborn) donor animals or to the culture conditions. PMID- 11032899 TI - Age-dependent differential responses of monoaminergic systems to high doses of methamphetamine. AB - Abuse of methamphetamine (METH) by adolescents is a major public health issue in the U.S.A. Because of the neurotoxic potential of METH, we examined the response of CNS monoaminergic systems in young (adolescent) animals [postnatal day (PND) 40] to high-dose treatments (10 mg/kg, four injections, 2-h intervals) of this drug and contrasted these effects to those seen in older (young adult) rats (PND 90). Consistent with previous reports, we observed that PND 40 animals did not manifest the long-term (7-day) deficits in extrapyramidal dopamine (DA) parameters observed in PND 90 rats. In contrast, METH-induced rapid (1-h) reduction in the activity of striatal DA transporters occurred in both age groups. In addition, both persistent (7-day) and rapid (1-h) deficits in serotonergic systems (measured as reductions in tryptophan hydroxylase activity) were observed in PND 40 and 90 rats. Age-related differences in METH-induced hyperthermia did not appear to be a principal cause for our observations; however, age-dependent pharmacokinetics of this drug might have contributed to the differential METH monoaminergic responses by PND 40 and 90 animals. PMID- 11032900 TI - RGS7 is palmitoylated and exists as biochemically distinct forms. AB - Regulator of G protein signaling (RGS) proteins are GTPase-activating proteins that modulate neurotransmitter and G protein signaling. RGS7 and its binding partners Galpha and Gbeta5 are enriched in brain, but biochemical mechanisms governing RGS7/Galpha/Gbeta5 interactions and membrane association are poorly defined. We report that RGS7 exists as one cytosolic and three biochemically distinct membrane-bound fractions (salt-extractable, detergent-extractable, and detergent-insensitive) in brain. To define factors that determine RGS7 membrane attachment, we examined the biochemical properties of recombinant RGS7 and Gbeta5 synthesized in Spodoptera frugiperda insect cells. We have found that membrane bound but not cytosolic RGS7 is covalently modified by the fatty acid palmitate. Gbeta5 is not palmitoylated. Both unmodified (cytosolic) and palmitoylated (membrane-derived) forms of RGS7, when complexed with Gbeta5, are equally effective stimulators of Galpha(o) GTPase activity, suggesting that palmitoylation does not prevent RGS7/Galpha(o) interactions. The isolated core RGS domain of RGS7 selectively binds activated Galpha(i/o) in brain extracts and is an effective stimulator of both Galpha(o) and Galpha(i1) GTPase activities in vitro. In contrast, the RGS7/Gbeta5 complex selectively interacts with Galpha(o) only, suggesting that features outside the RGS domain and/or Gbeta5 association dictate RGS7-Galpha interactions. These findings define previously unrecognized biochemical properties of RGS7, including the first demonstration that RGS7 is palmitoylated. PMID- 11032901 TI - 5-HT(1B) receptor-mediated regulation of serotonin clearance in rat hippocampus in vivo. AB - The 5-hydroxytryptamine (5-HT; serotonin) transporter (5-HTT) is important in terminating serotonergic neurotransmission and is a primary target for many psychotherapeutic drugs. Study of the regulation of 5-HTT activity is therefore important in understanding the control of serotonergic neurotransmission. Using high-speed chronoamperometry, we have demonstrated that local application of 5 HT(1B) antagonists into the CA3 region of the hippocampus prolongs the clearance of 5-HT from extracellular fluid (ECF). In the present study, we demonstrate that the 5-HT(1B) antagonist cyanopindolol does not produce this effect by increasing release of endogenous 5-HT or by directly binding to the 5-HTT. Dose-response studies showed that the potency of cyanopindolol to inhibit clearance of 5-HT was equivalent to that of the selective 5-HT reuptake inhibitor fluvoxamine. Local application of the 5-HT(1A) antagonist WAY 100635 did not alter 5-HT clearance, suggesting that the effect of cyanopindolol to prolong clearance is not via a mechanism involving 5-HT(1A) receptors. Finally, the effect of low doses of cyanopindolol and fluvoxamine to inhibit clearance of 5-HT from ECF was additive. These data are consistent with the hypothesis that activation of terminal 5 HT(1B) autoreceptors increases 5-HTT activity. PMID- 11032902 TI - Selective adrenergic/cyclic AMP-dependent switch-off of proteasomal proteolysis alone switches on neural signal transduction: an example from the pineal gland. AB - The molecular processes underlying neural transmission are central issues in neurobiology. Here we describe a novel mechanism through which noradrenaline (NA) activates its target cells, using the mammalian pineal organ as a model. In this neuroendocrine transducer, NA stimulates arylalkylamine N:-acetyltransferase (AANAT; EC 2.3.1. 87), the key enzyme regulating the nocturnal melatonin production. In rodents, AANAT protein accumulates as a result of enhanced transcription, but in primates and ungulates, the AANAT mRNA level fluctuates only marginally, indicating that other mechanisms regulate AANAT protein and activity. These were investigated in cultured bovine pinealocytes. AANAT mRNA was readily detectable in unstimulated pinealocytes, and levels did not change following NA treatment. In contrast, NA increased AANAT protein levels in parallel with AANAT activity, apparently through a cyclic AMP-mediated mechanism. Immunocytochemistry revealed that the changes in AANAT protein levels occurred in virtually all pinealocytes. Inhibition of AANAT degradation by proteasomal proteolysis alone was found to switch-on enzyme activity by increasing AANAT protein levels five- to 10-fold. Accordingly, under unstimulated conditions AANAT protein is continually synthesized and immediately destroyed by proteasomal proteolysis. NA appears to act via cyclic AMP to protect AANAT from proteolytic destruction, resulting in accumulation of the protein. These findings show that tightly regulated control of proteasomal proteolysis of a specific protein alone can play a pivotal role in neural regulation. PMID- 11032903 TI - Coordinated expression of beta-amyloid precursor protein and the putative beta secretase BACE and alpha-secretase ADAM10 in mouse and human brain. AB - To define the enzymes involved in the etiology of Alzheimer's disease, we compared in mouse and human brain the mRNA levels and cellular localization of the ubiquitous beta-amyloid precursor protein (beta-APP) with those of the putative alpha-secretases ADAM10 and ADAM17 and the beta-secretases BACE and BACE2. In situ hybridization performed in mice during prenatal and postnatal development and in adulthood revealed the coexpression of beta-APP, BACE, and ADAM10. The patterns of BACE2 and ADAM17 only partially overlapped with that of beta-APP. beta-APP, BACE, and ADAM10 mRNAs have also been detected by northern blot in human brain cortex of normal subjects and in Alzheimer's disease subjects. In situ hybridization performed using combined biotin- and radiolabeled riboprobes provided evidence for the coexpression of beta-APP with BACE and ADAM10 in human cortical neurons. Our data provide cytochemical evidence supporting the role of ADAM10 and BACE as authentic alpha- and beta-secretases. PMID- 11032904 TI - The role of reactive nitrogen species in secondary spinal cord injury: formation of nitric oxide, peroxynitrite, and nitrated protein. AB - To determine whether reactive nitrogen species contribute to secondary damage in CNS injury, the time courses of nitric oxide, peroxynitrite, and nitrotyrosine production were measured following impact injury to the rat spinal cord. The concentration of nitric oxide measured by a nitric oxide-selective electrode dramatically increased immediately following injury and then quickly declined. Nitro-L-arginine reduced nitric oxide production. The extracellular concentration of peroxynitrite, measured by perfusing tyrosine through a microdialysis fiber into the cord and quantifying nitrotyrosine in the microdialysates, significantly increased after injury to 3.5 times the basal level, and superoxide dismutase and nitro-L-arginine completely blocked peroxynitrite production. Tyrosine nitration examined immunohistochemically significantly increased at 12 and 24 h postinjury, but not in sham-control sections. Mn(III) tetrakis(4-benzoic acid)-porphyrin (a novel cell-permeable superoxide dismutase mimetic) and nitro-L-arginine significantly reduced the numbers of nitrotyrosine-positive cells. Protein-bound nitrotyrosine was significantly higher in the injured tissue than in the sham operated controls. These results demonstrate that traumatic injury increases nitric oxide and peroxynitrite production, thereby nitrating tyrosine, including protein-bound tyrosine. Together with our previous report that trauma increases superoxide, our results suggest that reactive nitrogen species cause secondary damage by nitrating protein through the pathway superoxide + nitric oxide peroxynitrite protein nitration. PMID- 11032905 TI - Reduced binding of protein phosphatase 2A to tau protein with frontotemporal dementia and parkinsonism linked to chromosome 17 mutations. AB - Coding region and intronic mutations in the tau gene cause frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). We have previously reported that ABalphaC, a major form of protein phosphatase 2A (PP2A) in brain, binds tightly to tau protein in vitro and is a major tau phosphatase in vivo. Using in vitro assays, we show here that the FTDP-17 mutations G272V, DeltaK280, P301L, P301S, S305N, V337M, G389R, and R406W inhibit by approximately 20-95% the binding of recombinant three-repeat and four-repeat tau isoforms to the ABalphaC holoenzyme and the AC core enzyme of PP2A. Reduction in binding was maximal for tau proteins with the G272V, DeltaK280, and V337M mutations. We also show that tau protein can be specifically coimmunoprecipitated with endogenous PP2A from both rat brain and transfected cell extracts. It is significant that, by using similar coimmunoprecipitation assays, we show that all FTDP-17 mutations tested, including the N279K mutation, alter the ability of tau to associate with cellular PP2A. Taken together, these results indicate that FTDP-17 mutations induce a significant decrease in the binding affinity of tau for PP2A in vivo. We propose that altered protein-protein interactions between PP2A and tau may contribute to FTDP-17 pathogenesis. PMID- 11032906 TI - Mice transgenic for the Huntington's disease mutation are resistant to chronic 3 nitropropionic acid-induced striatal toxicity. AB - Neuronal loss in Huntington's disease (HD) is seen first in the neostriatum. It has been suggested that impaired metabolism underlies this degeneration, as striatal vulnerability to excitotoxicity is increased by metabolic compromise. At 12 weeks of age, a transgenic mouse carrying the HD mutation (R6/2 line) has been shown to have an increased vulnerability to the mitochondrial toxin 3 nitropropionic acid (3-NP). However, in contrast, younger R6/2 mice appear to be less vulnerable than wild-type (WT) mice to the excitotoxins kainic acid and quinolinic acid (QA). In this study, we examine the possibility that the sensitivity of R6/2 mice to 3-NP might be age dependent. We treated young, symptomatic R6/2 mice with 3-NP and found that despite their progressive neurological phenotype, they were not more susceptible to 3-NP intoxication than their WT littermates. Further, fewer R6/2 than WT mice developed striatal lesions. We suggest that compensatory mechanisms exist in the R6/2 mouse brain that protect it against the toxic effect of the transgene and coincidentally protect against exogenous toxins such as 3-NP, QA, and kainic acid. The existence of similar compensatory mechanisms may explain why, in humans, HD is a late-onset disorder, despite early expression of the genetic mutation. PMID- 11032907 TI - Tissue plasminogen activator requires plasminogen to modulate amyloid-beta neurotoxicity and deposition. AB - Tissue plasminogen (plgn) activator (tPA) modulates neuronal death in models of stroke, excitotoxicity, and oxidative stress. Amyloid-beta (Abeta) appears central to Alzheimer's disease and is neurotoxic to neurons in vitro. Here, we evaluate tPA effects on Abeta toxicity. We report that tPA alone had no effect on Abeta toxicity. However, in combination with plgn, tPA reduced Abeta toxicity in a robust fashion. Moreover, the combined tPA and plgn treatment markedly inhibited Abeta accumulation. The addition of phenylmethylsulfonyl fluoride, a serine protease inhibitor, to a sample of tPA, plgn, and Abeta resulted in a marked reduction of Abeta degradation. We interpret the actions of tPA and plgn within the context of the ability of plasmin to degrade Abeta. PMID- 11032908 TI - Oxidative stress following traumatic brain injury in rats: quantitation of biomarkers and detection of free radical intermediates. AB - Oxidative stress may contribute to many pathophysiologic changes that occur after traumatic brain injury. In the current study, contemporary methods of detecting oxidative stress were used in a rodent model of traumatic brain injury. The level of the stable product derived from peroxidation of arachidonyl residues in phospholipids, 8-epi-prostaglandin F(2alpha), was increased at 6 and 24 h after traumatic brain injury. Furthermore, relative amounts of fluorescent end products of lipid peroxidation in brain extracts were increased at 6 and 24 h after trauma compared with sham-operated controls. The total antioxidant reserves of brain homogenates and water-soluble antioxidant reserves as well as tissue concentrations of ascorbate, GSH, and protein sulfhydryls were reduced after traumatic brain injury. A selective inhibitor of cyclooxygenase-2, SC 58125, prevented depletion of ascorbate and thiols, the two major water-soluble antioxidants in traumatized brain. Electron paramagnetic resonance (EPR) spectroscopy of rat cortex homogenates failed to detect any radical adducts with a spin trap, 5,5-dimethyl-1-pyrroline N:-oxide, but did detect ascorbate radical signals. The ascorbate radical EPR signals increased in brain homogenates derived from traumatized brain samples compared with sham-operated controls. These results along with detailed model experiments in vitro indicate that ascorbate is a major antioxidant in brain and that the EPR assay of ascorbate radicals may be used to monitor production of free radicals in brain tissue after traumatic brain injury. PMID- 11032909 TI - Effect of chronic psychogenic stress exposure on enkephalin neuronal activity and expression in the rat hypothalamic paraventricular nucleus. AB - This study tested the hypothesis that the activation pattern of enkephalinergic (ENKergic) neurons within the paraventricular nucleus of the hypothalamus (PVH) in response to psychogenic stress is identical whether in response to repeated exposure to the same stress (homotypic; immobilization) or to a novel stress (heterotypic; air jet puff). Rats were assigned to either acute or chronic immobilization stress paradigms (90 min/day for 1 or 10 days, respectively). The chronic group was then subjected to an additional 90-min session of either heterotypic or homotypic stress. A single 90-min stress session (immobilization or air jet) increased PVH-ENK heteronuclear (hn) RNA expression. In chronically stressed rats, exposure to an additional stress session (whether homotypic or heterotypic) continued to stimulate ENK hnRNA expression. Acute immobilization caused a marked increase in the numbers of Fos-immunoreactive and Fos-ENK double labeled cells in the dorsal and ventral medial parvicellular, and lateral parvicellular subdivisions of the PVH. Chronic immobilization caused an attenuated Fos response ( approximately 66%) to subsequent immobilization. In contrast, chronic immobilization did not impair ENKergic neuron activation within the PVH following homotypic or heterotypic stress. These results indicate that within the PVH, chronic psychogenic stress markedly attenuates the Fos response, whereas ENKergic neurons resist habituation, principally within the ventral neuroendocrine portion of the nucleus. This suggests an increase in ENK effect during chronic stress exposure. Homotypic (immobilization) and heterotypic (air jet) psychogenic stressors produce similar responses, including Fos, ENK-Fos, and ENK hnRNA, within each subdivision of the PVH, suggesting similar processing for painless neurogenic stimuli. PMID- 11032910 TI - Mitochondrial carbonic anhydrase in the nervous system: expression in neuronal and glial cells. AB - Carbonic anhydrase (CA) V is a mitochondrial enzyme that has been reported in several tissues of the gastrointestinal tract. In liver, it participates in ureagenesis and gluconeogenesis by providing bicarbonate ions for two other mitochondrial enzymes: carbamyl phosphate synthetase I and pyruvate carboxylase. This study presents evidence of immunohistochemical localization of CA V in the rodent nervous tissue. Polyclonal rabbit antisera against a polypeptide of 17 C terminal amino acids of rat CA V and against purified recombinant mouse isozyme were used in western blotting and immunoperoxidase stainings. Immunohistochemistry showed that CA V is expressed in astrocytes and neurons but not in oligodendrocytes, which are rich in CA II, or capillary endothelial cells, which express CA IV on their plasma face. The specificity of the immunohistochemical results was confirmed by western blotting, which identified a major 30-kDa polypeptide band of CA V in mouse cerebral cortex, hippocampus, cerebellum, spinal cord, and sciatic nerve. The expression of CA V in astrocytes and neurons suggests that this isozyme has a cell-specific, physiological role in the nervous system. In astrocytes, CA V may play an important role in gluconeogenesis by providing bicarbonate ions for the pyruvate carboxylase. The neuronal CA V could be involved in the regulation of the intramitochondrial calcium level, thus contributing to the stability of the intracellular calcium concentration. CA V may also participate in bicarbonate ion-induced GABA responses by regulating the bicarbonate homeostasis in neurons, and its inhibition could be the basis of some neurotropic effects of carbonic anhydrase inhibitors. PMID- 11032911 TI - Rat alpha-synuclein interacts with Tat binding protein 1, a component of the 26S proteasomal complex. AB - The alpha-synuclein gene, which encodes a brain presynaptic nerve terminal protein of unknown function, is linked to familial early-onset Parkinson's disease (PD). The finding that alpha-synuclein forms the major fibrillary component of Lewy bodies in brains of PD patients suggests that the two point mutations in alpha-synuclein (Ala(53)Thr, Ala(30)Pro) may promote the aggregation of alpha-synuclein into filaments. To address the role of alpha-synuclein in neurodegenerative diseases, we performed a yeast two-hybrid screen of a rat adult brain cDNA library using rat alpha-synuclein 2 (alphaSYN2). Here we report that alphaSYN2 interacts specifically with Tat binding protein 1, a subunit of the 700 kDa proteasome activator (PA700), the regulatory complex of the 26S proteasome and of the modulator complex, which enhances PA700 activation of the proteasome. PMID- 11032912 TI - Combination therapy with SCH58500 (p53 adenovirus) and cyclophosphamide in preclinical cancer models. AB - SCH58500 (ACN53) is a replication-deficient, recombinant adenovirus which expresses human p53 tumor suppressor. In preclinical models, SCH58500 has therapeutic efficacy against a wide range of human tumor types containing nonfunctional p53 and it has enhanced activity in combination with many chemotherapeutic drugs. However, the anti-tumor efficacy of SCH58500 combined with the DNA-damaging chemotherapeutic cyclophosphamide has not been previously reported. Cyclophosphamide did not enhance the activity of SCH58500 in three out of four human tumor xenograft models studied. Furthermore, combination therapy with SCH58500 and cyclophosphamide was not any better than single drug treatment in transgenic H-ras mice and in FVB mice bearing syngeneic MidT2-1 tumors. This is in sharp contrast to previous combination studies in these models where SCH58500 had enhanced efficacy when given with the farnesyl protein transferase inhibitor SCH66336, paclitaxel, cisplatin, cisplatin/paclitaxel, or doxorubicin. Further evaluation of this combination is required before it can be recommended for clinical trials in cancer patients. PMID- 11032913 TI - Survival of patients with cutaneous T-cell lymphoma after treatment with extracorporeal photochemotherapy. AB - Few studies have assessed the long-term outcome of patients with cutaneous T-cell lymphoma (CTCL) treated with extracorporeal photochemotherapy (ECP). Our objective was to assess the efficacy, safety, and survival of a cohort of patients with refractory CTCL in stages Ib to III who were treated with ECP. A retrospective study was performed. Twenty patients (19 male, 1 female) aged 38 to 87 years with CTCL of the mycosis fungoides type (n=17) and primary cutaneous Ki 1 lymphoma (n=2) were treated twice a month. Sixteen had an adjunctive treatment with interferon alpha (IFN alpha) s.c. 3 times a week in the maximal tolerable dosage (i.e. up to 21x106). A complete response was achieved in 10 patients, a partial response in three and a stable disease in seven patients (response rate 65.0%). The overall survival was 29.4+/-16.0 months, the event-free survival was 26.2+/-12.4 months, and the progression-free survival was 23. 4+/-12.2 months. Four patients died of causes unrelated to CTCL and two patient died of CTCL. Median survival time was 26 months. No severe side effects were noted. ECP is a safe alternative therapy for CTCL. In particular when combined with IFN alpha it can induce long-term remissions. PMID- 11032914 TI - A rapid immunoassay predicts BRCA1 and BRCA2 mutations in buccal cells. AB - We developed a rapid, non-invasive, and inexpensive, assay capable of identifying BRCA1 and BRCA2 (BRCA) mutations in buccal cells. To determine the predictive value of this immunoassay, a double blind study of 13 high risk individuals was conducted by two independent teams. As greater than 90% of BRCA mutations result in protein truncations, a diminished anti-carboxy immunoreactivity relative to anti-amino immunoreactivity was scored as predictive for mutation. Comparison to BRCA DNA analysis was undertaken. The positive and negative predictive values were 90% and 100% respectively (p<0.02), suggesting great promise as an inexpensive and rapid screen for BRCA mutations. PMID- 11032915 TI - Differential expression of matrix metalloprotease-7 in each component of uterine carcinosarcoma. AB - The aim of this study was to investigate the expression of matrix metalloprotease 7 (MMP-7) in each component of uterine carcinosarcoma. Surgical specimens of primary tumors of uterine carcinosarcomas were obtained from 13 patients. The carcinomatous component consisted of adenocarcinoma with or without squamous differentiation. The sarcomatous component consisted of spindle cell sarcoma, chondrosarcoma and rhabdomyosarcoma, either alone or in combination. The immunohistochemical expression of MMP-7 protein was examined using the avidin biotin peroxidase complex technique employing the anti-MMP-7 monoclonal antibody. Expression of MMP-7 protein was detected in 9 (69.2%) of 13 adenocarcinoma components, while no staining was observed in any of the sarcomatous components examined. There was a statistically significant difference of the positive rate for MMP-7 between epithelial components and sarcomatous components of uterine carcinosarcoma (p<0.01). In some cases, MMP-7 was abundantly expressed at the invasive front of adenocarcinomas. It is concluded that MMP-7 protein is differentially expressed between the carcinomatous component and the sarcomatous component of uterine carcinosarcoma. Each component of carcinosarcoma may have its own potential for invasion and metastasis. MMP-7 may contribute to the invasive nature or growth capacity of the carcinomatous component of uterine carcinosarcoma, while it may not have a relation to that of the sarcomatous components. PMID- 11032916 TI - Induction of apoptosis by Acanthopanax senticosus HARMS and its component, sesamin in human stomach cancer KATO III cells. AB - Antitumor effect of the stem bark of Acanthopanax senticosus HARMS (ASH) from Hokkaido (Japanese name: Ezoukogi) on human stomach cancer KATO III cells was investigated. The extract of the stem bark of ASH prepared with hot water was dissolved in distilled water and used for the assay of antitumor effect on the KATO III cells. The exposure of KATO III cells to ASH led to both growth inhibition and induction of apoptosis. Morphological change showing apoptotic bodies was observed in the cells treated with ASH. The fragmentation by ASH of DNA to oligonucleosomal-sized fragments that are characteristics of apoptosis was observed to be concentration- and time-dependent. We have investigated which component in ASH is effective on the induction of apoptosis. Among chlorogenic acid, syringaresinol di-o-beta-D glucoside, syringin, and sesamin, components of the n-butanol extract prepared from ASH, sesamin suppressed the growth and induced apoptosis in the cells. These findings suggest that growth inhibition by ASH results from the apoptosis induced by sesamin, a component of ASH. PMID- 11032917 TI - Cyclooxygenase-2 immunoreactivity in depressed-type colorectal adenomas and cancers. AB - In order to clarify the relationship of the cyclooxygenase-2 immunoreactivity and the progression of depressed-type colorectal adenomas and cancers, we examined cyclooxygenase-2 expression in depressed-type colorectal adenomas and cancers. Immunohistochemical staining was performed by the streptavidin-biotin method. Among 100 adenomas, depressed-type were found in 15, cyclooxygenase-2 expression was not detected in depressed-types, and lesion size in depressed-types did not differ from that in others. Among 100 submucosal invasive cancers, depressed-type were found in 14, cyclooxygenase-2 expression was not detected in depressed types, and lesion size in tumor with depressed-type did not differ from that in others. Among 100 cancers invading the muscularis propria, depressed-types in 15, cyclooxygenase-2 expression was not detected in depressed-types; these were significantly smaller lesions than those in others (p<0.05). Our results suggest that depressed-type cancers do not result from cyclooxygenase-2 expression. PMID- 11032918 TI - Induction of apoptosis by gallic acid in human stomach cancer KATO III and colon adenocarcinoma COLO 205 cell lines. AB - Antitumor effects of gallic acid on human stomach cancer KATO III cells and human colon adenocarcinoma COLO 205 cells were investigated. The exposures of KATO III and COLO 205 cells to gallic acid led to both growth inhibition and induction of apoptosis. Morphological changes showing apoptotic bodies were observed in both the cell lines treated with gallic acid. The fragmentations by gallic acid of DNA to oligonucleosomal-sized fragments, that are characteristics of apoptosis, were observed to be concentration- and time-dependent. These findings suggest that growth inhibitions by gallic acid of KATO III cells and COLO 205 cells result from the apoptosis induced by gallic acid. Thus, gallic acid might be a candidate drug for digestive gut cancer treatment to overcome the resistance to chemotherapeutic drugs. PMID- 11032919 TI - Increased expression levels of p53 correlate with good response to cisplatin based chemotherapy in non-small cell lung cancer. AB - In order to determine whether expression of the tumor suppressor gene p53 in non small cell lung cancer (NSCLC) correlates with chemotherapeutic response, resected tumors from 18 patients with recurrent lung cancer who had undergone complete resection and received chemotherapy after the initial tumor recurrence were subjected to p53 immunostaining. Histological examination of the resected tumors revealed 11 adenocarcinomas, 6 squamous cell carcinomas and one adenosquamous cell carcinoma. Group 1 was 50% (n=9) p53 immunopositive. All patients received cisplatin-based chemotherapy after recurrence. No patient in group 1 achieved response to chemotherapy whereas 2 and 3 in group 2 achieved complete and partial responses, respectively. The chemotherapy response rate of group 2 (56%) was significantly higher than that of group 1 (0%, p=0.009). The times to reoccurrence after tumor resection of group 2 was significantly better than that of group 1 (log-rank p=0.019, Wilcoxon p=0.042), and survival after chemotherapy of group 2 was also significantly better than that of group 1 (log-rank p=0.023, Wilcoxon p=0.034). It is suggested that high p53 expression levels in tumors correlate with both good response to cisplatin-based chemotherapy and good survival of patients with advanced NSCLC. PMID- 11032920 TI - c-erbB-2 protein expression and clinicopathologic features in colorectal cancer. AB - Previous reports showed that breast and gastric cancers overexpressing c-erbB-2 protein have a greater metastatic potential and worse prognosis than tumors in which this protein is not overexpressed. The present study was undertaken to examine the significance of c-erbB-2 protein expression as a prognostic factor in colorectal cancer. Protein expression was examined immunohistologically in colorectal cancer tissue from 149 patients without distant metastasis, from 38 patients with liver metastasis, and from 18 patients with lung metastasis. The c erbB-2 protein-positive rate was significantly higher in cases with lymphatic vessel invasion in the primary tumor, but it did not correlate with lymph node metastases. Expression of c-erbB-2 did not correlate with any other histologic feature (histologic type, depth of tumor invasion, venous vessel invasion, or the clinical stage). The positive rate in the primary lesion was significantly higher in cases with liver metastasis than in cases without liver metastasis, the positive rate was significantly higher in the hepatic than in the primary lesions. The expression of c-erbB-2 protein in colorectal cancer tissue correlates closely with liver metastasis but not with lymphatic or lung metastases. PMID- 11032921 TI - Bleomycin, actinomycin-D, and cisplatin treatment of ovarian germ-cell malignancies contributes to reducing adverse drug reactions. AB - Sixteen ovarian germ-cell malignancy (OGCM) patients between 1983 and 1999 were randomly divided into the BEP group (n=6; bleomycin, etoposide and cisplatin) and the BAP group (n=10; bleomycin, actinomycin-D and cisplatin). The patients were evaluated for adverse drug reactions (ADRs) based on severity-grading of the National Cancer Institute Common Toxicity Criteria. The ADRs in the BAP group were milder than in the BEP group, as seen in regard to alopecia (p=0.0126), low hemoglobin (p=0.0147), and decreased neutrophil count (p=0.0197). The five-year survival rate was 87.5% in the BAP group and 83.3% in the BEP group, and the difference was not significant (p=0.5954). BAP therapy is likely to be more beneficial for OGCM patients than BEP therapy, because ADRs are reduced with no difference in outcome. PMID- 11032922 TI - A preoperative protocol for the prevention of infection in children with tunnelled right atrial catheters. AB - The use of central venous lines has come to be widely accepted by children with cancer and their families. However, attendant infection is a cause of considerable morbidity. Coagulase-negative staphylococci, the predominant aerobic species on the skin, are now the commonest cause of catheter-related bacteremia. We introduced a protocol to reduce the colonization of the skin at the catheter insertion site. Antiseptic skin scrubs, with 4% chlorhexidine gluconate, were performed on the neck and anterior chest the night before and again on the morning of the surgical procedure. A single dose of cephalothin (or vancomycin for penicillin-allergic patients) was administered IV immediately before the operation. Compared to the 12 month period prior to initiation of this protocol, the rate of infections (occurring within 30 days of catheter placement) in the 3.5 year period of intervention dropped from 8 to 4.9 per 1,000 catheter days. The proportion of infections that were staphylococcal was reduced from 93 to 63% and the proportion of non-ports removed within 30 days of placement fell from 45 to 0%. Despite these changes, the major contribution to improved infection control appeared to be the use of an increased proportion of ports (a rise from <10 to almost 60%). PMID- 11032923 TI - Kinetics of plasma platinum in a hemodialysis patient receiving repeated doses of cisplatin. AB - The pharmacokinetics of cisplatin were studied in a 46-year-old hemodialysis patient treated for uterine cancer with weekly cisplatin. Hemodialysis was performed immediately before, one-hour after, and two-hours after cisplatin administration for two consecutive cycles each. The concentrations of total and free platinum were measured. The decay curve of both total platinum and free platinum showed a biphasic pattern characterized by an alpha-phase and a beta phase, the same as in non-hemodialysis patients. There were no differences between the individual cycles in the peak platinum concentration or the decay pattern. The plasma platinum concentration decreased to below detection limit 3 months after the final dose. PMID- 11032924 TI - Breast-conservation treatment for patients with ductal carcinoma in situ. AB - Fourteen cases with symptomatic ductal carcinoma in situ (DCIS) were treated with breast-conservation treatment intensified with endocrine therapy. Nine of 14 patients with palpable mass had tumor detected on mammography. CT, ultrasonography, and MRI were able to detect linear and/or spotty lesion or enhancement suggesting DCIS. Whereas these findings were not particular to DCIS, the combination of these modalities would be useful in deciding the extent of resection for DCIS. There was no patient selection for breast-conservation treatment in our department. All patients received tangential and boost radiation, and were treated with endocrine therapy using anti-estrogen drugs. The reason that nine cases had close margins (<5 mm) might be on account of the treatment including lumpectomy with 1 cm of surgical margin. In spite of their margin status, no local or systemic failure was experienced, and the cosmetic results of most patients were rated as excellent or good. Therefore, our breast conservation treatment intensified with systemic therapy is thought to be adequate for patients with symptomatic DCIS. Six of eight cases who received preoperative treatment containing endocrine therapy with or without CAF chemotherapy showed a decrease in tumor size. Preoperative treatment may effect the microinvasion and/or breast tissue surrounding a DCIS tumor. PMID- 11032925 TI - Indoor air pollution and pulmonary adenocarcinoma among females: a case-control study in Shenyang, China. AB - Factors that affect the risk of lung adenocarcinoma among females were investigated in Shenyang, China, using a population-based case-control study design. A total of 72 new cases, ages 35-69, diagnosed with incident, primary pulmonary adenocarnoma, were collected between April 1991 and December 1995, and were 1:1 age-matched with healthy females randomly selected from the general population. A questionnaire covering demographics, diet/nutritional preferences and cooking habits, living conditions, family history of cancer, sources of indoor/outdoor/occupational pollution, exposure to ETS from spousal smoking, workplace exposure, and exposure during childhood, history of menstruation and pregnancy, was given to each subject in a structured in-person interview given by trained field workers. Univariate analysis was performed on the data collected. The results showed that cooking fumes, family history of lung cancer, economic status, and number of live births and intake of vitamin E were risk factors significantly associated with adenocarcinoma of the lung. In particular, exposure to different levels of cooking fumes, an indoor air pollutant, increased the odds ratio of lung adenocarcinoma by 1.33, 7.33 and 1.67, respectively (trend p=0.006). Another important risk factor was family history of lung cancer, which gave an OR of 7.65 (95% CI, 0.90-169.84). Intake of beta-carotene from vegetables and fruit offered protection against lung adenocarcinoma, giving an OR of 0.28 (95% CI, 0.12-0.69). These results were confirmed by multivariable logistic regression analysis. PMID- 11032926 TI - The role of 18F-fluoro-2-deoxy-D-glucose positron emission tomography (18F-FDG PET) in the diagnosis of recurrence and lymph node metastasis of cervical cancer. AB - The usefulness of positron emission tomography with 2-[18F]-fluoro-2-deoxy-D glucose (FDG-PET) in diagnosing recurrence and lymph node metastasis of uterine cervical cancer was evaluated as a preliminary study. The FDG uptake of 13 cervical cancer patients, including 4 patients with recurrence, was evaluated. The FDG uptake of the cervices of 5 patients who underwent PET studies for non cervical tumors were used as control. In 2 patients who had undergone radical hysterectomy, we compared the diagnosis of lymph node metastasis based on FDG-PET and magnetic resonance imaging. The FDG uptake in the 9 primary cervical cancers was greater than that in normal cervices (p=0.025). Examination of the 4 recurrent tumors revealed a higher FDG uptake than that in normal cervices (p=0.0022). Metastatic lesions in lymph nodes showed an accumulation of FDG on the PET scan. In conclusion, FDG-PET may be an effective diagnostic tool for detecting recurrence and lymph node metastasis of uterine cervical cancers. PMID- 11032927 TI - Antitumor effects of fotemustine and busulfan against a human neuroblastoma xenograft. AB - We examined whether or not fotemustine, a new nitrosourea derivative, and busulfan, an agent already clinically used, are effective against human neuroblastoma, using a human neuroblastoma xenograft model designated TNB9. The maximum inhibition rate (MIR) of fotemustine against the TNB9 model was 44.6% with a total dose of fotemustine of 75 mg/kg, indicating that fotemustine is not effective against TNB9. The MIR of busulfan against the same model was 26.7% when a total dose of 135 mg/kg was administered orally to nude mice. Busulfan was also suspended in carboxy-methylcellulose, and was administered intraperitoneally. The MIRs were 19.4% and 36. 4% when busulfan was administered intraperitoneally at a total dose of 40 mg/kg and 60 mg/kg, respectively. The total doses of 40 mg/kg and 60 mg/kg did not show any adverse effects on mice, but were found to be ineffective against TNB9, indicating that busulfan might not be an effective chemotherapeutic agent against human neuroblastoma. PMID- 11032928 TI - Evaluation of the UBC test in the urine of healthy individuals, patients with benign disorders and urinary bladder cancer. AB - Using the UBC test, the specificity, sensitivity and prognostic information were evaluated in patients with recently diagnosed transitional cell carcinoma (TCC) and in a control group consisting of apparently healthy individuals and individuals with benign disorders. Frozen urine samples from the 485 individuals in the control group and 100 newly diagnosed TCC patients were analyzed with the UBC test, specific for epitopes on cytokeratin fragments released from the urothelial cells. All the samples were analyzed and corrected for creatinine. No significant concentration difference was found between males and females (p=0.65) and there was no age dependent relation. The median concentration for the entire control group was estimated at 3.7 microg/g and the 95th percentile was calculated at 53.0 microg/g. The apparently healthy individuals in the control group had a median value of 3.4 microg/g with a 95th percentile of 24.3 microg/g. An increased frequency of elevated UBC concentrations was found in some benign disorders e.g., anemia, thyroid disorders, diabetes mellitus, hyperlipemia, urosepsis and cystitis. Patients with superficial tumors exhibited a 66% sensitivity (at 95% specificity), and the UBC concentrations did not differ statistically (p=0.16) from those patients with muscle invasive lesions with a 52% sensitivity. When the UBC concentrations were related to histopathological grade, a significant concentration difference (p<0.004) was found between low grade tumors (sensitivity 41%) and high grade tumors (sensitivity 72%). Survival analysis showed that patient with muscle invasive tumors, high-grade tumors and high UBC concentrations have a significantly reduced survival (five-year survival was estimated to 30%, 35% and 30% respectively) compared to patients with superficial tumors, low-grade tumors or low UBC concentrations (five-year survival, 60%, 85% and 75% respectively). The UBC test showed good accuracy and repeatability. Clinically the test could assist in tumor grading and the detection of recurrent disease, which in turn could assist in treatment selection for the individual patient and possibly improve prognosis. PMID- 11032929 TI - The results of radiation therapy for glottic carcinoma: prognostic significance of tumor size in laryngoscopic findings. AB - Seventy-five patients with carcinoma of the true vocal cord were treated with curative intent by radiotherapy at Kure National Hospital between 1977 and 1997. All tumors were classified according to stage (T1, T2 and T3), degree of differentiation (well-differentiated, moderately differentiated, poorly differentiated and unknown), and size (very small, small and large). All patients were treated using a cobalt-60 unit to a total dose that ranged from 56 Gy to 64 Gy (mean, 60 Gy). The mean treatment time was 44 days (range, 38-50 days). The local control and ultimate local control rates at 5 years were 75.8% and 91.4% for all patients, respectively. Univariate analysis showed that tumor size (p=0.0350) and stage T (p=0.0416) affected local control. In multivariate analysis, tumor size was the only significant factor that affected local control (p=0.0012). The most significant factor that affected local control was tumor size in laryngoscopic findings. Tumor size was inversely related to local control. PMID- 11032930 TI - Role of regenerative mucosa in primary gastric cancer treated by chemotherapy. AB - We radiographically evaluated the response of primary lesions to chemotherapy in 101 patients with inoperable advanced gastric cancer. The median survival time in the 50 responders (49.5%) was 379 days, as compared with only 210 days in the 51 non-responders (50.1%) (p<0. 001). Morphologic changes of the gastric mucosa were assessed endoscopically. Among the changes observed, we focused on regenerative mucosa and its relation to response and survival. Regenerative mucosa was seen in 56 (55%) of the 101 patients. The regenerative mucosa was examined endoscopically and classified into four patterns: flat, radial, granular and cobble-stone. Biopsy specimens were taken from the regenerative mucosa and examined histopathologically. The response to chemotherapy was found to be closely associated with the incidence of regenerative mucosa. In particular, a flat or radial pattern significantly correlated with a positive response to treatment (p=0.004) and was associated with significantly longer survival than granular or cobblestone pattern or no regenerative mucosa (p<0.001). Flat or radial pattern was also associated with low tumor-positive rate of biopsy specimens. We conclude that endoscopic evaluation of regenerative mucosa in patients with inoperable advanced gastric cancer who receive chemotherapy can provide useful information in the assessment of response and prognosis. PMID- 11032931 TI - Adenosine receptor antagonism causes inhibition of angiogenic activity of human ovarian cancer cells. AB - Angiogenesis, new blood vessels development, is an important process involved in ovarian cancer growth and metastasis. Adenosine is a known mediator of angiogenesis in hypoxic tissues. Theobromine, adenosine receptor antagonist, exerts antiangiogenic properties in many types of tumors in Balb/c mice cutaneous angiogenesis assay. The aim of the present study was to evaluate the mechanism of its action. We showed that theobromine inhibits angiogenic activity of ovarian cancer cells as well as CD45 positive lymphocytes isolated from peritoneal ascitic fluid of ovarian cancer patients. Using synthetic adenosine receptor antagonists: 8-phenyl-theophylline and 8-cyclopentyl-1,3 dipropylxantine we established that antiangiogenic properties of theobromine are dependent on its interaction with A2 adenosine receptor. Our observations were confirmed in full suspensions of ascitic cells as well as in isolated cancer cells and CD45 lymphocytes. We postulate that A2 receptor antagonism may diminish angiogenesis induced by hypoxia in different cancer tissues and may find a place in future cancer therapy. PMID- 11032932 TI - Bright loop appearance; a characteristic ultrasonography sign of early hepatocellular carcinoma. AB - Ultrasonography (US) and computed tomography (CT) are the most effective screening methodologies for hepatocellular carcinoma (HCC). In our US screening, 20% of small HCC nodules less than 20 mm in diameter were detected as hyperechoic tumors. Among these hyperechoic HCC nodules, we have often observed (BL) which is defined as hypoechoic nodules in the hyperechoic tumor. In this study, we report that the BL is a sign of dedifferentiation of early stage of HCC with fatty change by US. From 1994 to 1998, we performed tumor targeting needle biopsy in 938 hepatic nodular lesions. Among them, 284 nodules <20 mm in diameter, histologically diagnosed as HCC, were studied. BL is defined as a hyperechoic tumor containing a hypoechoic nodule >4 mm in diameter by US. Among 284 nodules, well, moderately and poorly differentiated HCC were 183 (64.4%), 100 (35.2%) and 1 (0.4%), respectively. On US, hypoechoic, isoechoic, and hyperechoic nodules were 188 (66.2%), 32 (11.3%) and 64 (22.5%), respectively. Forty-seven nodules of 64 hyperechoic HCC nodules <20 mm in diameter, 47 nodules (73.4%) showed fatty changes. Of 64 hyperechoic HCC nodules, we recognized 22 nodules (34.4%) as BL. The proportion of BL type hyperechoic nodules increased with the tumor size. Two hyperechoic nodules followed by US changed to BL with tumor enlargement. Histologic examination of a resected HCC with BL showed that hyperechoic HCC nodule represented well-differentiated HCC with fatty change and inner hypoechoic lesion represented moderately differentiated HCC without fatty change. In US screening for HCC, BL was often observed in HCC nodules from 11 to 20 mm in diameter. Histologic examination revealed that BL of HCC on US was associated with tumor progression and indicated dedifferentiation showing moderately differentiated HCC in well-differentiated HCC with fatty change. PMID- 11032934 TI - Bolus and infusional 5-fluorouracil combined with cisplatin in advanced gastric cancer. AB - 5-Fluorouracil (5-FU) is the main drug used in the treatment of advanced gastric cancer. Combination chemotherapy is not always superior to 5-FU alone, especially when biomodulators are also administered. In an attempt to exploit all the cytotoxic mechanisms of 5-FU, we carried out a pilot study with a double route of administration of 5-FU (intravenous bolus and continuous infusion) and a multiple modulation of 5-FU by methotrexate (MTX), 6S-leucovorin (6S-LV) and cisplatin (CDDP). A group of 30 patients affected by advanced gastric cancer was treated with MTX 50 mg/m2 and 5-FU 400 mg/m2 as an i.v. bolus on day 1, followed by a 5 day i. v. continuous infusion of 5-FU 600 mg/m2/day and 6S-LV 100 mg/m2/day; on day 3 CDDP 100 mg/m2 was also administered. The regimen was repeated every 4 weeks. Six partial responses (20+/-14. 3%), 12 stable diseases (40+/-17.5%) and 12 progression (40+/-17.5%) were observed in an intent-to-treat analysis. Median survival was 7 months. All responding patients had performance status 0-1. Grade 3-4 toxicity was mainly gastrointestinal, but grade 3-4 anemia and leucopenia were also recorded. The schedule has low activity. The use of different modulators and way of administration of 5-FU does not provide advantages in advanced gastric cancer. PMID- 11032933 TI - Unrecognized hepatic steatosis and non-alcoholic steatohepatitis in adjuvant tamoxifen for breast cancer patients. AB - Adjuvant tamoxifen has become the treatment of choice against estrogen receptor positive breast cancer. Adverse effects are rarely observed and since symptoms of hepatic steatosis, non-alcoholic steatohepatitis and cirrhosis are usually negligible, such effects are not well characterized despite large cohort studies of adjuvant tamoxifen. This issue remains to be systematically studied. The present study consisted of 136 breast cancer patients treated with or without tamoxifen. Patients had laboratory tests once each month and underwent abdominal computed tomography (CT) annually for 5 years. The extent of hepatic steatosis was assessed by CT as the liver/spleen ratio. While receiving adjuvant tamoxifen, 40 of 105 patients developed hepatic steatosis (liver/spleen ratio <0.9) without obvious changes in body mass index. Twenty-one had a liver spleen ratio of <0.5, whereas none of the 31 patients treated without tamoxifen had a ratio <0.9 or <0.5 (p<0.0001 and p<0.0001, respectively). Hepatic steatosis was recognized in 35 of the 40 patients within the first 2 years of receiving adjuvant tamoxifen and 21 of the 40 had increased transaminase levels. Liver biopsy revealed NASH in 6 of 7 patients among the 21 with a liver/spleen ratio of <0.5. A subset of individuals given adjuvant tamoxifen developed progressive hepatic steatosis without significant changes in the body mass index. We suggest a liver/spleen ratio of <0.5 as a criterion upon which liver biopsy should be recommended since NASH frequently occurred in such patients. PMID- 11032935 TI - Pleural effusion and pulmonary injury as an unusual complication to chemotherapy in non-small cell lung cancer patients. AB - We report the appearance of pleural effusion, or pulmonary failure after chemotherapy, followed by tumor reduction, in a small number of patients. Five hundred and fifty-four patients with lung cancer have undergone chemotherapy at our Institute during the last ten years. Three patients with non-small cell lung cancer (NSCLC), with locally advanced disease, exhibited an unusual consequence following cytotoxic drug treatment. Two patients with NSCLC had pleural effusion which improved within 2-3 weeks, together with tumor reduction, which allowed the continuation of treatment. One patient had pulmonary failure with pleural effusion and recovered within two weeks. Two of the three patients had positive cytology for cancer cells in the fluid. All three patients achieved partial remission with no repetition of the complication. The patients' recovery, response to treatment and the tumor reduction suggest that this complication was not due to disease progression. PMID- 11032936 TI - Prognostic value of numerical aberrations of chromosome 17 in differentiated gastric cancer: evaluation by multivariate regression analysis. AB - Using in situ hybridization with a chromosome-specific DNA probe, we investigated the numerical aberrations of chromosome 17 in differentiated gastric cancers. In 75 cancers that invaded the submucosal or deeper layer, the frequency of an increased hybridized signal (hyperdiploid nuclei) on chromosome 17 was significantly associated with lymph node metastasis. Multivariate analysis showed that the classification of numerical aberration (disomy and polysomy) based on the percentage of hyperdiploid nuclei was an independent prognostic factor for the postoperative survival of 58 patients with invasive gastric cancer treated with radical gastrectomy. Thus, numerical aberrations of chromosome 17 may be a useful prognostic indicator of differentiated gastric cancer. PMID- 11032937 TI - Antagonistic effect of EGF against TGF beta1 on transformed phenotype and tropomyosin expression of human lung carcinoma A549 cells. AB - TGF beta1 suppressed the colony forming ability of human lung carcinoma cell line, A549 cells in a dose-dependent manner. Accompanying these changes, the levels of the expression of high-molecular weight tropomyosin (TM) isoforms, especially TM1 and TM2, were significantly increased in the TGF beta1-treated A549 cells in a dose-dependent manner. When A549 cells were treated with TGF beta1 (2.5 pM) and EGF (1 nM) simultaneously, anchorage-independent growth of A549 cells was restored. In these cells, the expression of high-molecular weight TM decreased. The results suggest that the levels of the expression of high molecular weight TM may be involved in antagonistic effects of EGF and TGF beta1 on the anchorage-independent growth of A549 cells. PMID- 11032938 TI - Evaluation of inflammatory cytokines in malignant and benign pleural effusions. AB - We measured the levels of inflammatory cytokines interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-alpha) in pleural effusions and serum in 65 consecutive patients: 32 with malignant pleural effusion (MPE) (group A), and 33 with inflammatory benign pleural effusion (BPE) (group B). Serum levels of 15 healthy individuals served as control. Concentrations of IL-1alpha were higher in serum compared to pleural fluid in both groups (47.1+/-33.9 vs. 25.9+/-1.7 fmol/ml, p<0.001, in group A; and 39.9+/ 30.9 vs. 25.4+/-16.3 fmol/ml, p<0.02, in group B). Similarly, concentrations of IL-1beta and IL-2 were significantly higher in serum compared to pleural fluid in both groups. In contrast, IL-6, IL-8 and TNF-alpha were found at high concentration in MPE in comparison to serum IL-6: 171.8+/-60.4 vs. 7. 2+/-7 fmol/ml (p<0.001), IL-8: 1175.15+/-2385.6 vs. 285.2+/-187.2 pg/ml (p<0.05), TNF alpha: 204.9+/-82.9 vs. 79.4+/-31.9 fmol/ml (p<0. 001). Similarly, pleural concentrations of IL-6, IL-8 and TNF-alpha were higher in BPE patients in comparison to serum IL-6: 124.3+/-56. 2 vs. 8.6+/-6.4 fmol/ml (p<0.001) IL-8: 2109.2+/-4121.5 vs. 291. 6+/-197.9 pg/ml (p<0.02), TNF-alpha: 183.8+/-28.2 vs. 86.2+/-23.9 fmol/ml (p<0.001). These data suggest that IL-6, IL-8 and TNF-alpha might be secreted locally at the site of active disease both in benign and malignant pleural effusions. PMID- 11032939 TI - Serum concentration of pyridinoline cross-linked carboxyterminal telopeptide of type I collagen in patients with metastatic breast cancer. AB - The serum concentration of pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) was examined in 83 patients with metastatic breast cancer. ICTP levels were significantly higher in patients with bone metastases than in those without bone metastasis. In patients with bone metastasis, significantly higher ICTP levels were observed in those with multiple lesions than in those with a solitary lesion and these levels reflected therapeutic response. Sequential monitoring of ICTP revealed that this elevation was correlated with disease progression. Combined with imaging studies, monitoring of ICTP appears to offer additional information for detection of bone metastasis and evaluation of therapeutic response to bone metastasis. PMID- 11032940 TI - Structure, expression and mutational analysis of the hBRAG gene on 10q in the frequently deleted region in human endometrial cancer. AB - We previously reported that chromosome arm 10q is one of the target regions of allelic loss in human endometrial cancer. To identify the gene in this region responsible for endometrial cancer, we further characterized this region and localized the hBRAG gene. The function of hBRAG has not yet been fully studied, and there is the possibility that this gene works as a tumor suppressor. This gene consist of 7 exons and 6 introns encoding 503 amino acids; all the introns start with GT and end with AG in agreement with the GT-AG rule. Expression of this gene was studied by Northern hybridization and suppressed expression was observed in one (SK-UT-1B) of the six endometrial cancer cell lines. Mutation analysis in 38 primary EC tissues and six EC cell lines disclosed no genetic alterations. The genomic structure of hBRAG elucidated in this study should contribute to the future analysis of the hBRAG gene. PMID- 11032941 TI - Long-term survival after immunochemotherapy for juvenile colon cancer with peritoneal dissemination: a case report. AB - A 20 year-old man was hospitalized with an abdominal mass and abdominal distension. Investigations resulted in a diagnosis of ileus caused by advanced colon cancer with peritoneal dissemination to the pouch of Douglas. Palliative surgery was performed to relieve bowel obstruction and debulk the tumor. Histopathological examination showed that the tumor was a mucinous adenocarcinoma invading the serosa without lymph node metastasis. Ascites collected during the operation was diagnosed as class V. Administration of PSK (3.0 g/day) and UFT (600 mg/day) as adjuvant immunochemotherapy was started postoperatively to achieve tumor dormancy. He has been followed as an outpatient for 2.5 years with no ascites or abdominal symptoms. PMID- 11032942 TI - Papillary carcinoma in thyroglossal duct remnants: presentation of four cases and decision procedure for prophylactic thyroid gland dissection. AB - Papillary carcinoma in thyroglossal duct remnants is a rare and usually unexpected finding. It is controversial whether or not prophylactic thyroid gland dissection is necessary in such circumstances. We present our experience of four cases. Based on this, a consideration of published risk factors, and evaluation of the likelihood of a primary versus metastatic origin of the malignancy, we present a therapeutic decision procedure. When the thyroid is normal, the patient presents low-risk factors for thyroid cancer, and there is evidence that the malignancy is primary, removal of all thyroglossal duct remnants by the Sistrunk procedure is sufficient. PMID- 11032943 TI - In vitro MRS and HPLC studies on human renal cell carcinomas. AB - Samples from ten patients with renal cell carcinomas (RCC) were examined by 1H MRS and high performance liquid chromatography (HPLC) measurements. Twenty samples, ten from the tumor and ten from the surrounding healthy renal region were obtained. MRS yielded information on renal osmolytes which can be considered markers of physiological kidney function. The marked decrease of these osmolytes is a hallmark of cancer. Moreover, HPLC measurements disclosed the amino acid pattern of both healthy and neoplastic tissues. The Glu and Ea content had statistical significance. GSH was present in tumor tissues only. PMID- 11032944 TI - Intestinal intussusception in adults, treat it like a cancer. AB - Intussusception in adult is a rare condition. Pathologic process is identifiable in 90% of cases. Emergency clinical diagnosis is generally difficult and CT scan typical features are of use. Surgery is always necessary to treat bowel necrosis and to perform pathologic examination. Diagnosis of cancer must be in mind and lymphadenectomy must be done. We present two recent cases of intussusception in adults due to adenomatosis polyposis and Meckel's diverticulum. PMID- 11032945 TI - The time of tumor cell division and death depends on the site of growth. AB - We show here, for the first time, in two very different murine tumors, a mammary one (ectoderm) and a lung one (endoderm), that: tumors have day/night differences of spontaneous apoptosis additional to the well-known circadian rhythm of mitosis. The times of maximal and minimal mitosis and apoptosis changed for a tumor cell line when growing in different organs (as metastasis) or anatomical sites. Both tumor lines, have identical circadian curves when growing in a specific organ or anatomical site. The peaks of apoptosis match with the valleys of mitosis and vice versa. PMID- 11032946 TI - L-dopa-resistant parkinsonism syndrome following cerebral radiation therapy for neoplasm. AB - A bradykinetic form of parkinsonism, unresponsive to levo-dopa therapy developed in four patients two to eight weeks after completion of external beam irradiation (39.2 Gy to 59.4 Gy) of their intracranial neoplasm. In the absence of other causative factors, we relate the movement disorder to radiation-induced changes within the basal ganglia. At post-mortem examination one patient had putamenal gliosis and thickened vessels with loss of nigral neurons. PMID- 11032947 TI - Microsatellite alterations and K-ras, TGFbetaRII, IGFRII and bax mutations in sporadic cancers of the gastrointestinal tract. AB - DNA was analyzed from 57 sporadic gastrointestinal tumors (34 pancreatic cancers, 23 colon tumors) and cognate normal tissues to verify whether mutations at coding sequences were associated with microsatellite instability (MSI). Genomic instability was present in 41% (14/34) of pancreatic samples and in 26% (6/23) of colon cancers previously tested by six microsatellite markers. The tumors included 37 cases showing no MSI; 15 cases with MSI at only 1 locus and 5 cases with MSI at 2 or more loci. All the samples were screened for mutations in genes containing repeated tracts in their coding sequences (TGFbetaRII, IGFRII and bax) and in codon 12 of the K-ras oncogene. Furthermore, loss of heterozygosity (LOH) at NM23.H1 locus was tested, 17/34 (50%) pancreatic tumors and 6/23 (26%) colon cancers showed mutations in codon 12 of K-ras; allelic loss of NM23. H1 locus was found in 6/18 (33%) informative colon tumors and in no pancreatic cancers. The TGFbetaRII, IGFRII and bax genes were altered in 3 (13%), 1 (4%) and 3 (13%) out of 23 colon tumors respectively, but no mutation was detected in pancreatic cancers. Mutations in the repeated nucleotide stretches within the coding sequences of TGFbetaRII, IGFRII and bax genes were found only in colon tumors with a high unstable phenotype (more than 3 microsatellite loci altered). PMID- 11032948 TI - Chemotherapeutic evaluation of Celecoxib, a cyclooxygenase-2 inhibitor, in a rat mammary tumor model. AB - Epidemiological and experimental studies have shown that non-steroidal anti inflammatory drugs (NSAIDs) reduce the relative risk of human cancer, including breast cancer. Recently, research studies in our laboratories have shown that the selective cyclooxygenase-2 (COX-2) blocker, Celecoxib, given daily in the diet, significantly inhibited the induction of rat mammary tumors by 7, 12 dimethylbenz(a)anthracene (DMBA). These studies were extended to evaluate Celecoxib for its effectiveness as an antineoplastic agent in this rat mammary tumor model. We examined the growth inhibitory effects of Celecoxib, given daily in the diet, on the volume and the number of established mammary tumors, vis-a vis the cancer load (CL). Tumors continued to grow actively in control rats fed chow diet only. In contrast, the Celecoxib-supplemented diet (1500 mg/kg diet) significantly decreased the size of the mammary tumors in rats over the 6 week treatment period, resulting in an average reduction in tumor volume of approximately 32%, relative to the baseline volume (p<0.04). At the end of the 6 week treatment period, average tumor volume was 1.45 cm3 and 0.13 cm3 in the control and Celecoxib treated rats respectively. Tumor regression occurred in 90% of the rats. In addition, new tumors continued to emerge in the control group, in contrast to their significantly decreasing numbers in the Celecoxib treated group over the same time period (p<0.05). These results indicate that Celecoxib has significant antineoplastic activity, in addition to its anticarcinogenic effects. PMID- 11032949 TI - Immunohistochemical study of fatty acid synthase in ovarian neoplasms. AB - In order to evaluate the prognostic significance of fatty acid synthase (FAS), the key enzyme in fatty acid biosynthesis, in ovarian benign and malignant tumors 98 patients with ovarian masses were immunohistochemically evaluated. Histologic features of the lesions and expression for FAS were statistically associated with clinical data and disease-free survival over a 5-year follow-up period. The mean age of patients was 65 years. TNM surgical stages were I in 31.8%, II in 28.2% and III in 40% of cases. The 5-year disease-free survival of the 85 malignant common epithelial ovarian tumors was 47.1% while 35.3 of patients died with active disease. Seventy-eight (79.6%) out of 98 cases expressed FAS in the neoplastic epithelial cells and 42 (42.9%) in the non-neoplastic stromal cells. Statistical analysis revealed FAS expression and stage of the disease to be significantly correlated to recurrence and survival rates only in the malignant common epithelial tumors. Moreover, multivariate analysis showed FAS expression (p=0.05) and tumor stage (p=0.03) to be independent prognostic predictors. FAS is a reliable predictor of recurrence and disease-free survival in common epithelial ovarian tumors along with neoplastic stage. Clinical data, other histologic subtypes and grading were not statistically significant. PMID- 11032950 TI - Orofacial viral infections in the immunocompromised host. AB - Orofacial viral infections are common in immunocompromised patients. Herpes simplex virus (HSV) infections are the most common. Varicella-Zoster virus (VZV) infections are less common, but usually more severe. Epstein-Barr virus (EBV) may produce ulcers, lymphoproliferative syndromes or oral hairy leukoplakia (HL). Human herpes virus 6 (HHV 6) may be the etiology of recurrent aphthous stomatitis, while human herpes virus 7 (HHV 7) has an unclear role. Human herpes virus 8 (HHV 8) is the etiologic agent of Kaposi sarcoma. Human papilloma viruses (HPVs) cause warts, papillomas and epithelial hyperplasia. Molluscum contagiosum is caused by a poxvirus. All herpes viruses (HSV, VZV, CMV, EBV, HHV 6, HHV 7, HHV 8) and many HPVs are associated with neoplasias. Research is ongoing to clarify the role of other viruses in the development of infections and lesions in the orofacial area. PMID- 11032951 TI - Cathepsin B levels in urine from bladder cancer patients. AB - There is accumulating evidence that cysteine proteinase activity plays an important role in cancer cell invasion and metastasis. Previously we demonstrated that cathepsin B (CB) plasma activity is increased in patients with transitional bladder cancer (TCC). In this work we have attempted to determine whether urine CB protein levels could be used as tumor marker in bladder cancer patients. Urine CB levels were evaluated employing a dot blot method, in 30 patients with TCC, 21 patients successfully treated from TCC without evidence of disease at the moment of urine collection (NED) and in 30 healthy volunteers. The median value (Md) of the control group was 3.8 microg CB/ml. Significantly higher urine CB values (Md: 5.9 microg/ml) were found in the TCC group. A high CB value was also found in the NED group (5.0 microg/ml). Urine CB values over the 5.2 microg/ml (cut-off point) were observed in 63% of TCC patients, 48% of NED and 8% of the control group. Only 4% NED patients had CB values over 13.0 microg/ml while 33% of TCC patients surpassed this value. Thus, urine CB might be a potential marker for transitional bladder cancer diagnosis. PMID- 11032952 TI - Melatonin and colon carcinogenesis. IV. Effect of melatonin on proliferative activity and expression of apoptosis-related proteins in the spleen of rats exposed to 1,2-dimethylhydrazine. AB - The suppression of 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis by melatonin was previously demonstrated. The objective of the present work was to evaluate histologically and immunohistochemically the splenic immune response to the induced cancer and to melatonin. Spleens from rats, either untreated, injected with DMH, fed with melatonin or treated with both carcinogen and melatonin, were studied. The exposure to the carcinogen and the consequential carcinogenesis resulted in splenic changes that reflected the insufficiency of the immune response, as manifested in significant reduction of the white pulp and the simultaneous expansion of the red pulp. The effects of melatonin on most splenic components were inverse to those of DMH. The anti-carcinogenic properties of melatonin were evidenced from the reversal of the inhibitory effects of DMH, especially when the densities of lymphocytes in different parts of the spleen were compared. The combined treatment of the rats with DMH and melatonin resulted in the expansion of the splenic zones by 106% to 125%, compared to those from DMH treated rats, and the numbers of CD8+ lymphocytes and Fas-positive cells increased sharply. Therefore we conclude that anti-carcinogenic effects of melatonin are related to activation of several elements of the host's lymphatic system. PMID- 11032953 TI - Mini-review: computational structure-based design of inhibitors that target protein surfaces. AB - Finding drugs that inhibit protein-protein interactions is usually difficult. While computer-aided design is used widely to facilitate the drug discovery process for protein targets with well-defined binding pockets, its application to the design of inhibitors targeting a protein surface is very limited. In this mini-review we address two aspects of this issue: firstly, we overview the current state of design methodology for inhibitors specifically targeting protein surfaces, and secondly, we briefly outline recent advances in computational methods for structure-based drug design. These methods are closely related to protein docking and protein recognition, the difference being that in ligand design, ligands are built on a fragment-by-fragment basis. A novel scheme of computational combinatorial ligand design developed for the design of inhibitors that interfere with protein-protein interaction is described in detail. Current applications and limitations of this methodology, as well as its future prospects, are discussed. PMID- 11032954 TI - Molecular descriptors in chemoinformatics, computational combinatorial chemistry, and virtual screening. AB - Many contemporary applications in computer-aided drug discovery and chemoinformatics depend on representations of molecules by descriptors that capture their structural characteristics and properties. Such applications include, among others, diversity analysis, library design, and virtual screening. Hundreds of molecular descriptors have been reported in the literature, ranging from simple bulk properties to elaborate three-dimensional formulations and complex molecular fingerprints, which sometimes consist of thousands of bit positions. Knowledge-based selection of descriptors that are suitable for specific applications is an important task in chemoinformatics research. If descriptors are to be selected on rational grounds, rather than guesses or chemical intuition, detailed evaluation of their performance is required. A number of studies have been reported that investigate the performance of molecular descriptors in specific applications and/or introduce novel types of descriptors. Progress made in this area is reviewed here in the context of other computational developments in combinatorial chemistry and compound screening. PMID- 11032955 TI - Targeting of phage display vectors to mammalian cells. AB - Phage display libraries offer a strategy to isolate peptide ligands to target proteins and to define potential interaction sites between proteins. Recent studies have indicated a novel utility for phage display in that bacteriophage engineered to express peptide ligands to specific cell surface receptors are internalized by mammalian cells. Thus, reporter genes such as green fluorescent protein and lacZ harbored in the phage genome can be delivered to mammalian cells using targeting peptides displayed on the surface of phage. There is also the possibility to generate novel types of peptide libraries expressed intracellularly using a phage capable of inducing expression of its coding genes in human cells. PMID- 11032956 TI - High throughput mutation screening by automated capillary electrophoresis. AB - Molecular diagnosis of complex inherited disorders, population screening of genetic diseases, studies of the genetic basis of variable drug response (pharmacogenetics) as well as discovery and investigation of new drug targets (pharmacogenomics) involve screening for mutations in multiple DNA samples. Furthermore, the development of a third generation of the human genome map, based on single nucleotide polymorphisms (SNPs), requires screening for allelic variants through all of the three billion basepairs in the human genome. Thus, the need for high throughput mutation screening methods is great and is rapidly increasing. Traditional methods for mutation screening often involve slab-gel electrophoresis analyses which are laborious and difficult to automate. However, recent developments in capillary electrophoresis systems for DNA fragment analysis have made fully automated mutation screening possible and have dramatically increased the possible sample throughput. This review describes the recent advances in capillary electrophoresis of DNA and summarize the various methods for mutation screening based on this technique. PMID- 11032957 TI - High throughput and global approaches to gene expression. AB - In the past several years, a new set of technologies based on whole genome analysis have revolutionized the study of gene expression. These microarray or "gene chip" technologies, which arose out of the development of large-scale sequencing approaches, are now coming into increasing use, generating a far greater volume of data than the data representing the sequences themselves. This review focuses on the current state of development of these technologies, and the available approaches to manage and analyze the information they generate. The applicability of this technology to general problems in biomedicine is also discussed. PMID- 11032958 TI - Forecasting roles of combinatorial chemistry in the age of genomically derived drug discovery targets. AB - Genomics has caused an explosion in the number of potential therapeutic targets with varying degrees of validated pathophysiology. Among the first applications of combinatorial chemistry in genomics-driven drug discovery is the search for surrogate ligands or substrates. In the event that no surrogate is found for molecular assays, more exotic functional screens in whole cells or model organisms are used. Protein-protein interaction mapping by yeast and mammalian two-hybrid systems dominates empirical functional genomics, and this will lead to a bias for screening projects targeting this type of interaction. Drug discovery for protein-protein interactions has a poor track record, and this will challenge prevailing views on the design of combinatorial libraries. Genomics based on structural homology will yield many putative kinases, receptors, enzymes, transporter proteins, ion channels and GPCRs. Most of these projects will require new surrogate agonists, ligands or substrates, and then pharmaceutically useful agonists or antagonists will need to be found. Again, combinatorial chemistry might be essential to these studies. Given the need to screen hundreds of targets at great risk of irrelevance to pathophysiology, combined with the challenge of finding surrogate or natural ligands for these new targets, there is an urgent need for efficiency. Different groups are addressing these concerns by developing biologically-driven combinatorial libraries in order to achieve a higher density of bioactivity. Early efforts in this regard will be described. PMID- 11032959 TI - Fluorescence polarization is a useful technology for reagent reduction in assay miniaturization. AB - The use of fluorescence polarization (FP) has increased significantly in the development of sensitive and robust assays for high throughput screening of chemical compound libraries during the past few years. In this study, we show that FP is a useful assay miniaturization technology for reagent reduction during high throughput screening. We developed and optimized several FP assays for binding to estrogen receptor alpha and two protein kinases with an assay volume of 100 microl. Without any re-optimization, a consistent signal window was maintained in 384- or 1536-well format when the assay volume varied from 2.5-100 microl at constant concentrations of all assay components. In contrast, the signal window decreased with decreasing assay volume at constant reagent concentration in the protein kinase C scintillation proximity assay (SPA) and prompt fluorescence assay. In addition, the effect of evaporation on the signal window was minimal for the FP assays. Our study suggests that FP is superior to SPA and prompt fluorescence in terms of reagent reduction in the miniaturized assay format. PMID- 11032960 TI - Structural approaches to explain the selectivity of COX-2 inhibitors: is there a common pharmacophore? AB - The identification and characterisation of the isoenzyme cyclooxygenase 2 (COX-2) stimulated investigations to develop efficient non-steroidal anti-inflammatory drugs with reduced side effects compared to standard NSAIDs. This review will focus on the structural features needed to achieve COX-2 selectivity. Five structural classes can be identified together with a class bearing little or no resemblance to one another in their molecular structure. The most interesting point is the very distinct structure/activity relationship. On the one hand only minor modifications to a particular compound induce a drastic change in its COX selectivity and on the other hand the structural prerequisites in terms of molecular shape, lipophilicity, electron density, flexibility, polarity and H bonding dynamics allow a wide range of diversity. PMID- 11032961 TI - COX-2 in brain and spinal cord implications for therapeutic use. AB - Following the discovery of inducible COX-2 in arthritic joint fluid and immunocompetent cells a revolution in the field of antiinflammatory treatment was expected. The detection of a constitutive COX-2 in the kidney, in stomach and central nervous system destroyed this hypotheses. Further experiments in animal models were done to elucidate the role of the constitutive COX-2 in different physiological and pathophysiological states. In central nervous system was shown that the constitutive COX-2 is the predominant isoform of cyclooxygenases in brain and spinal cord and is highly regulated by different mediators. After experimental induction of peripheral inflammation a significant induction of COX 2 gene, protein expression and synthesis of prostaglandins in the spinal cord was detected. It was concluded that COX-2 is strongly involved in pain mediation processing in the spinal cord. The detection of COX-2 in the brain endothelial cells and its role in fever led to new insights of development and time course of temperature elevation. Probably, the use of selective COX-2 inhibitors decreases fever more effective than classical antipyretics. Furthermore, newer results show a role of COX-2 in differentiation and maturation processes in brain. These findings implicate new ways for the treatment of Alzheimer's disease and other degenerative brain disorders. Clinical and experimental results with selective COX-2 inhibitors show a better safety profile than non-selective COX inhibitors. The clinical use after drug registration will be decide on the further role of this new class of drugs in analgesic/antiinflammatory therapy and on new fields of clinical use. PMID- 11032962 TI - Cyclooxygenase-2 inhibition and side-effects of non-steroidal anti-inflammatory drugs in the gastrointestinal tract. AB - Inhibition of prostaglandin biosynthesis via inhibition of the fatty acid cyclooxygenase (COX) is the mechanism of action of non-steroidal anti inflammatory drugs (NSAIDs). This results in an inhibition of the inflammatory and pain-producing activities of prostaglandins at a site of tissue injury but also in inhibition of prostaglandin production in the gastrointestinal tract (GI) and platelets, i.e. sites where endogenous prostaglandins are possibly involved in control of physiological functions. The discovery of two COX isoenzymes, COX-1 and COX-2, and the detection of their separate function and regulation, has initiated the search for new and putatively more selective inhibitors of prostaglandin biosynthesis. Specifically, selective inhibitors of COX-2 were developed in order to improve the anti-inflammatory and analgetic specificity and potency of the compounds and to reduce side-effects in the GI tract. Available experimental and clinical data of selective COX-2 inhibitors, including flosulide, celecoxib or rofecoxib, suggest improved gastric tolerance as compared to conventional, non-selective NSAIDs. However, experimental evidence suggests that both, the analgetic and anti-inflammatory action of COX-inhibitors, might also require inhibition of COX-1. COX-2-selective compounds at anti-inflammatory doses might have other side-effects, and for example reduce vascular prostacyclin production. Evidence is accumulating that COX-2 might not only be considered as a putatively detrimental enzyme but rather a highly regulated enzyme that also contributes to tissue protection and is even constitutively expressed in healthy human stomach mucosa. This paper reviews some of these newer aspects of COX-2 selective inhibitors in clinical use and discusses their possible benefits and risks. PMID- 11032963 TI - COX-2 inhibition, apoptosis, and chemoprevention by nonsteroidal anti inflammatory drugs. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) have as their common mechanism the inhibition of cyclooxygenase (COX) enzymes, of which two isoforms (COX-1 and COX 2) exist. The effect of NSAIDs on chemoprevention and tumor regression has been shown in animal models, epidemiologic studies, and in treatment of patients. The exact biochemical and cellular mechanisms underlying each of these phenomena is only partially understood. Processes that have been recently implicated as being important include the inhibition of tumor cell growth, prevention of angiogenesis, and induction of apoptosis in neoplastic cells. PMID- 11032964 TI - COX-2 selectivity and inflammatory processes. AB - Increasing amounts of experimental and clinical data support the role of selective cyclooxygenase (COX)-2 inhibition in anti-inflammatory processes and the involvement of COX-1 inhibition in the side effects associated with non steroidal anti-inflammatory drug use. This review will focus on the differences in the structure of the COX-1 and COX-2 molecules, particularly the active site and how they are bound by various NSAIDs to achieve COX-2 selectivity. This COX-2 selectivity will then be characterized in pharmacological assays in vitro and in animal models in vivo. Finally, clinical information available for this new class of selective inhibitors will be discussed. PMID- 11032965 TI - Role of cyclooxygenases in angiogenesis. AB - Angiogenesis is the process by which new blood vessels are formed. This process supports normal physiology as well as contributes to progression of disease. Progressive rheumatoid arthritis and growth of tumors are two pathologies to which angiogenesis contributes. In arthritis, we know that prostaglandins (PGs) and the enzyme cyclooxygenase-2, which catalyses prostaglandin production, are inflammatory mediators. These mediators are involved in rheumatoid arthritis and cancer-induced angiogenic processes. We discuss, herein, recent findings on the expression of cyclooxygenases in both rheumatoid arthritis and human cancer, and the links between COX-2, PGs, and angiogenesis. We also propose a model for the possible mechanistic interaction of the various cell types involved in angiogenesis. PMID- 11032967 TI - Recent developments of rebeccamycin analogues as topoisomerase I inhibitors and antitumor agents. AB - DNA topoisomerases are essential for the survival of prokaryotic and eukaryotic organisms. Topoisomerases inhibitors, due to their capacity to induce DNA breaking, can exhibit interesting antitumor properties. While there are many potent antitumor agents which target topoisomerase II, relatively few families of specific topoisomerase I inhibitors have been identified. The present review describes a new family of topoisomerase I inhibitors, analogues of the bacterial metabolite rebeccamycin. These compounds possess an indolocarbazole chromophore onto which is attached a sugar residue. Important structure-activity relationships studies in this series have helped to understand the role of the carbohydrate moiety which is absolutely necessary for topoisomerase I poisoning, the influence of the stereochemistry (alpha or beta) of its linkage to indole, the influence of the functionalities and substitutions on the sugar moiety and on the aromatic framework have been investigated. In addition to their action on DNA, rebeccamycin analogues may inhibit the SR kinase activity of topoisomerase I and therefore constitute a unique family of topoisomerase I poisons quite different from the well known camptothecins. PMID- 11032966 TI - A survey of calpain inhibitors. AB - Calpain is unique among the cysteine protease family of enzymes in that it combines thiol protease activity with calmodulin-like activity. Despite its wide spread distribution the exact physiological function(s) of calpain is yet to be deciphered. The enzyme is however, implicated in a number of pathophysiological conditions. Due to the potential of calpain as a therapeutic target a number of inhibitors have been described for the enzyme. In this article we have grouped calpain inhibitors into those derived from natural sources, and those derived from chemical synthesis. Additionally, an overview of functional groups that have been used as warheads of calpain inhibitors is presented along with a discussion of the structure activity relationship studies of the address region of peptidyl calpain inhibitors. Recent work in this area has led to a better understanding of the structural requirements for tight binding of inhibitors to the active site of calpain. A discussion of peptidomimetic calpain inhibitors, nonpeptide calpain inhibitors, and selectivity of some calpain inhibitors are also presented. The recent disclosure of the crystal structure of a nonpeptide calpain inhibitor bound to a hydrophobic pocket on the calcium-binding domain of calpain has opened the door to future development of potent cell permeable nonpeptide calpain inhibitors. PMID- 11032968 TI - Inhibitors of cyclin-dependent kinases as anti-cancer therapeutics. AB - Initiation, progression, and completion of the cell cycle are regulated by various cyclin-dependent kinases (CDKs), which are thus critical for cell growth. Tumour development is closely associated with genetic alteration and deregulation of CDKs and their regulators, suggesting that inhibitors of CDKs may be useful anti-cancer therapeutics. Indeed, early results suggest that transformed and normal cells differ in their requirement for e.g. cyclin/CDK2 and that it may be possible to develop novel antineoplastic agents devoid of the general host toxicity observed with conventional cystostatic drugs. Numerous active-site inhibitors of CDKs have been studied; the main limitation with these ATP antagonists is kinase specificity for CDKs. However, screening of compound collections, as well as rational design based on enzyme-ligand complex crystal structures, are now yielding pre-clinical candidates, particularly certain purine and flavonoid analogues, with impressive potency and selectivity. Natural CDK inhibitors (CKIs), e.g. the tumour suppressor gene products p16(INK4), p21(WAF1), and p27(KIP1), form the starting point for the design of mechanism-based CDK inhibitors. A number of these small proteins have been dissected and inhibitory lead peptides amenable to peptidomimetic development have been identified. Conversion of these peptides into pharmaceutically useful molecules is greatly aided by the recent elucidation of CKI/CDK crystal and solution structures. Additional interaction sites on CDKs being exploited for the purposes of inhibitor design include: phosphorylation/dephosphorylation sites, macromolecular substrate binding site, CKS regulatory subunit binding sites, cyclin-binding site, cellular localisation domain, and destruction box. Finally, progress has recently been made in the application of antisense technology in order to target CDK activity. PMID- 11032969 TI - Crisis periods and apoptotic commitment: death imprints? AB - Crisis periods in development are critical periods of cell death that have long been suggested as "epigenetic crises" which are central to normal and abnormal embryogenesis. Under in-vitro culture conditions, there are similar crisis periods or Hayflick limits of culture senescence. Epigenetic modulations from CpG methylation coupled to DNA replication provide an alternate timing mechanism to the telomere/telomerase biological clock. Physiological cell death in both development and in in-vitro isolates is primarily apoptotic. Arguments of divergent apoptotic death commitments as caspase dependent and independent pathways seem to suggest that there is no possibility of a global life and death signal. However recent reports implicating CpG specific cleavage in apoptosis implies that the powerful imprint mark that silences genes, protects genes from nuclease restriction, and modulates chromatin conformations, could provide a common commitment pathway of convergence in the death cascade. If the imprint mark were central to the apoptotic commitment, then apoptosis is Lamarckian not Darwinian. PMID- 11032970 TI - Chemical and pharmacological aspects of heteroaryl-nitrones. AB - Radical induced oxidative damage is extremely harmful to tissues and organs due to molecular modifications brought to polyunsaturated membrane lipids, proteins and nucleic acids. Oxidative stress is believed to be one of the pathophysiological mechanisms that operate in neurodegenerative disorders such as cerebral ischemias, amyotrophic lateral sclerosis, Parkinson s and Alzheimer s diseases. Nitrones oppose oxidative challenges by virtue of their ability to trap very rapidly oxygen or carbon centered radicals thus generating nitroxide radical species which are more stable and biochemically less harmful than the original radical. However the operational mechanism of nitrones might also go beyond direct scavenging of radicals. The chemical and pharmacological properties of nitrones depend strongly on the connectivity as well as on the type and position of the substituents in the compound's architecture. Heteroaryl-nitrones are known, but except for a few cases (for example pyridyl-nitrones) no particular attention has been given to this class of molecules. The following review is a survey of the literature reports on this subject from 1980 to 1999. The structures were classified according to the heterocyclic substituent on the nitrone double bond, and documented pharmaceutical features were emphasized. Whenever possible heteroaromatic and related aromatic nitrones were compared. PMID- 11032971 TI - Adenosine receptor ligands-recent developments part I. Agonists. AB - Developments in the field of adenosine receptor (AR) agonists of the past years are presented and discussed. Four different AR subtypes, A(1), A(2A), A(2B), and A(3), have been cloned from different species including the human receptors. Recombinant ARs expressed in permanent mammalian cell lines have found wide application in the screening of new ligands. Considerable differences are observed among data from different laboratories, using recombinant receptors for the assays. Reevaluation of compounds at all four receptor subtypes has shown that agonists that were believed to be selective for either A(1) or A(2A) ARs may be potent A(3) agonists and thus, non-selective. Potent and selective agonists for two of the AR subtypes, A(1) and A(3), have been developed. Truly selective A(2A) AR agonists, however, are presently not available. Potent or selective A(2B) agonists are still lacking. Since the treatment with AR agonists may lead to fast desensitization of the receptors, partial agonists, and indirect AR agonists, such as adenosine kinase inhibitors, or allosteric enhancers of adenosine binding, are being developed as site- and event-specific agents. PMID- 11032972 TI - Antitumour metallocenes: structure-activity studies and interactions with biomolecules. AB - The metallocene dihalides are a relatively new class of small, hydrophobic organometallic anticancer agents that exhibit antitumour properties against numerous cell lines including leukemias P388 and L1210, colon 38 and Lewis lung carcinomas, B16 melanoma, solid and fluid Ehrlich ascites tumours and several human colon and lung carcinomas transplanted into athymic mice. Titanocene dichloride 1 has been the most widely studied metallocene and the drug is currently in phase II clinical trials. Formation of metallocene-DNA complexes has been implicated in the mechanism of antitumour properties of the metallocenes, as both titanocene dichloride 1 and vanadocene dichloride 2 inhibit DNA and RNA synthesis, and titanium and vanadium accumulate in nucleic acid-rich regions of tumour cells. However, in contrast to the well characterized platinum-based anticancer drugs, the active species responsible for antitumour activity in vivo has not been identified and the mechanism whereby irreparable DNA damage and/or structural modification of DNA or other cellular targets occurs is poorly understood. This review will focus on recent studies that have been carried out in order to identify the biologically active species and more fully understand the molecular level mechanism of action of the metallocene dihalides. Studies with nucleotides, oligonucleotides, DNA and proteins including topoisomerases, protein kinase C and transferrin have provided important insight into potential cellular transport mechanisms and the interaction of metallocenes with biomolecular targets. New structure activity studies including the design of hydrolytically stable metallocenes and the preparation of highly water soluble amino acid analogues have not led to improved anticancer activity of titanocene dichloride 1. The vastly different chemical and hydrolytic stability of each of the metallocenes points to a unique mechanism of action of each metallocene in vivo. PMID- 11032973 TI - The analysis and crystallographic characterization of 20-hydroxyecdysone. AB - 20-Hydroxyecdysone (20E) is an insect molting hormone that is also widely spread in various plants. Many chromatographic methods can be used to identify and/or determine 20E content in samples of biological origin and various spectroscopic methods serve to identify its structural elements. We have utilized X-ray crystallography to reveal the stereostructures of 20E. Our data demonstrates that 20E exists in two different crystalline forms that are both orthorhombic modifications. One form is homo-molecular, with a limited freedom of internal rotation of the side chain around the C23-C24 bond and the other, which is a clathrate formed with methanol and water, which minimize the conformational freedom of the side chain. PMID- 11032974 TI - Modulation of amphetamine-induced striatal dopamine release by ketamine in humans: implications for schizophrenia. AB - BACKGROUND: Recent brain imaging studies have indicated that schizophrenia is associated with increased amphetamine-induced dopamine release in the striatum. It has long been hypothesized that dysregulation of subcortical dopamine systems in schizophrenia might result from a failure of the prefrontal cortex (PFC) to adequately control subcortical dopaminergic function. The activity of midbrain dopaminergic neurons is regulated, in part, by glutamatergic projections from the PFC acting via glutamatergic N-methyl-D-aspartate (NMDA) receptors. The goal of this study was to test the hypothesis that a pharmacologically induced disruption of NMDA transmission leads to an increase in amphetamine-induced dopamine release in humans. METHODS: In eight healthy volunteers, we compared striatal amphetamine induced (0.25 mg/kg) dopamine release under control conditions and under sustained disruption of NMDA transmission induced by infusion of the noncompetitive NMDA antagonist ketamine (0.2 mg/kg intravenous bolus followed by 0.4 mg/kg/hour intravenous infusion for 4 hours). Amphetamine-induced dopamine release was determined with single photon emission computed tomography, as the reduction in the binding potential (BP) of the radiolabeled D(2) receptor antagonist [(123)I]IBZM. RESULTS: Ketamine significantly enhanced the amphetamine induced decrease in [(123)I]IBZM BP, from -5.5% +/- 3.5% under control conditions to -12. 8% +/- 8.8% under ketamine pretreatment (repeated-measures analysis of variance, p =.023). CONCLUSIONS: The increase in amphetamine-induced dopamine release induced by ketamine (greater than twofold) was comparable in magnitude to the exaggerated response seen in patients with schizophrenia. These data are consistent with the hypothesis that the alteration of dopamine release revealed by amphetamine challenge in schizophrenia results from a disruption of glutamatergic neuronal systems regulating dopaminergic cell activity. PMID- 11032975 TI - Cortical bcl-2 protein expression and apoptotic regulation in schizophrenia. AB - BACKGROUND: The etiology of schizophrenia remains unknown; however, a role for apoptosis has been hypothesized. Bcl-2 is a potent inhibitor of apoptosis and also exerts neurotrophic activity in the central nervous system (CNS). Bcl-2 expression is increased in the CNS of several neurodegenerative disorders. Given that schizophrenia has certain features of a limited neurodegenerative disorder, it was hypothesized that cortical Bcl-2 expression is increased in schizophrenia. METHODS: Postmortem temporal cortex was obtained from the Stanley Foundation Neuropathology Consortium with matched control, schizophrenic, bipolar, and depressed subjects. Bcl-2 protein was measured by enzyme-linked immunoassay (ELISA) and Western blot. Primary analysis was limited to schizophrenia versus control subjects. RESULTS: The ELISA demonstrated 25% less Bcl-2 protein in schizophrenia (p =.046), supported by Western blot results. A secondary analysis of schizophrenic and bipolar subjects revealed twofold higher mean Bcl-2 in antipsychotic-treated versus neuroleptic-naive subjects. CONCLUSIONS: Contrary to our hypothesis, cortical Bcl-2 was reduced in schizophrenia. This supports the notion that schizophrenia is not a classic neurodegenerative disorder; however, less Bcl-2 protein may signal neuronal vulnerability to proapoptotic stimuli and to neuronal atrophy. Also, the association between neuroleptic exposure and higher Bcl-2 levels could underlie the favorable long-term outcomes of patients who receive maintenance antipsychotic treatment. PMID- 11032976 TI - Abnormalities in the thalamus and prefrontal cortex during episodic object recognition in schizophrenia. AB - BACKGROUND: Many patients with schizophrenia demonstrate memory deficits. We studied patterns of brain activity during episodic recognition of new and previously seen three-dimensional objects. METHODS: We used (15)O positron emission tomography to study regional cerebral blood flow in eight normal subjects and nine patients with schizophrenia during a visual object recognition task. RESULTS: In comparison with control subjects, patients with schizophrenia showed less regional cerebral blood flow increases in the pulvinar region of the right thalamus and the right prefrontal cortex during the recognition of new objects and significantly greater left prefrontal cortex regional cerebral blood flow increases during the recognition of previously seen objects. Patients with schizophrenia exhibited alarm rates to new objects similar to those of control subjects, but significantly lower recognition rates for previously seen objects. CONCLUSIONS: Schizophrenia is associated with attenuated right thalamic and right prefrontal activation during the recognition of novel visual stimuli and with increased left prefrontal cortical activation during impaired episodic recognition of previously seen visual stimuli. This study provides further evidence for abnormal thalamic and prefrontal cortex function in schizophrenia. PMID- 11032977 TI - Chronic valproate treatment increases expression of endoplasmic reticulum stress proteins in the rat cerebral cortex and hippocampus. AB - BACKGROUND: Sodium valproate is a highly effective treatment for bipolar disorder, but its mechanism of action remains poorly understood. We recently found with differential display polymerase chain reaction that valproate regulates the expression of the endoplasmic reticulum stress protein GRP78 in the rat cerebral cortex. In our study, we investigated the effect of this drug on the other members of the endoplasmic reticulum stress protein family, GRP94 and calreticulin, and we studied the brain regional distribution of GRP78, GRP94, and calreticulin. METHODS: Immunohistochemistry was used to measure protein levels of GRP78, GRP94, and calreticulin after treatment with sodium valproate (300 mg/kg, intraperitoneal) in specific rat brain regions. RESULTS: We report here that chronic treatment with valproate also increased expression of other members of the endoplasmic reticulum stress protein family, such as GRP94 and calreticulin. The brain regional distribution of these changes was similar for all three proteins, with marked increase detected in the frontal cortex, parietal cortex, and CA1 region of the hippocampus. CONCLUSIONS: Because GRP78, GRP94, and calreticulin possess molecular chaperone activity and bind Ca(2+) in the endoplasmic reticulum, the pharmacologic action of valproate may involve one or more of these processes. PMID- 11032978 TI - Associated disturbances in calcium homeostasis and G protein-mediated cAMP signaling in bipolar I disorder. AB - BACKGROUND: Evidence of extensive cross-talk between calcium (Ca(2+))- and cAMP mediated signaling systems suggests that previously reported abnormalities in Ca(2+) homeostasis in bipolar I (BP-I) patients may be linked to disturbances in the function of G proteins that mediate cAMP signaling. METHODS: To test this hypothesis, the beta-adrenergic agonist, isoproterenol, and the G protein activator, sodium fluoride (NaF), were used to stimulate cAMP production in B lymphoblasts from healthy and BP-I subjects phenotyped on basal intracellular calcium concentration ([Ca(2+)](B)). cAMP was measured by radioimmunoassay and [Ca(2+)](B) by ratiometric fluorometry with fura-2. RESULTS: Isoproterenol- (10 microM) stimulated cAMP formation was lower in intact B lymphoblasts from BP-I patients with high [Ca(2+)](B) (>/= 2 SD above the mean concentration of healthy subjects) compared with patients having normal B lymphoblast [Ca(2+)](B) and with healthy subjects. Although basal and NaF-stimulated cAMP production was greater in B lymphoblast membranes from male BP-I patients with high versus normal [Ca(2+)](B), there were no differences in the percent stimulation. This suggests the differences in NaF response resulted from higher basal adenylyl cyclase activity. CONCLUSIONS: These findings suggest that trait-dependent disturbances in processes regulating beta-adrenergic receptor sensitivity and G protein mediated cAMP signaling occur in conjunction with altered Ca(2+) homeostasis in those BP-I patients with high B lymphoblast [Ca(2+)](B). PMID- 11032979 TI - Neuropsychologic impairments in bipolar and unipolar mood disorders on the CANTAB neurocognitive battery. AB - BACKGROUND: Cognitive deficits associated with mood disorders, especially bipolar disorder, have been the focus of limited systematic investigation. METHODS: We tested 35 bipolar (21 in depressed state and 14 in mixed or manic state) and 58 nonbipolar depressed consecutively admitted young adult inpatients and 51 matched healthy individuals on the Cambridge Neuropsychological Test Automated Battery, a computerized neurocognitive battery. RESULTS: The mixed/manic bipolar patients demonstrated robust deficits in episodic and working memory, spatial attention, and problem solving. In contrast, depressed bipolar and nonbipolar patients demonstrated impairments only in episodic memory. CONCLUSIONS: Neuropsychologic findings with the Cambridge Neuropsychological Test Automated Battery indicate widely distributed deficits in cognitive domains subserved by temporal, parietal, and frontostriatal systems in bipolar patients during mixed/manic states of illness. Significant deficits in bipolar and nonbipolar depressed patients were restricted to episodic memory, suggesting a more selective dysfunction in mesial temporal lobe function during episodes of depression. These findings highlight the different cognitive profiles of mania and depression, demonstrate similar patterns of neuropsychologic deficits in bipolar and nonbipolar depression, and point to a need for further research investigating the characteristics, causes, course, and treatment of severe cognitive deficits associated with mixed/manic phases of bipolar disorder. PMID- 11032980 TI - Proton magnetic resonance spectroscopy of human basal ganglia: response to cocaine administration. AB - BACKGROUND: Proton magnetic resonance spectroscopy was used to determine the effects of intravenous cocaine or placebo administration on human basal ganglia water and metabolite resonances. METHODS: Long echo time, proton magnetic resonance spectra of water and intracellular metabolites were continuously acquired from an 8-cm(3) voxel centered on the left caudate and putamen nuclei before, during, and after the intravenous administration of cocaine or a placebo in a double-blind manner. RESULTS: Cocaine, at both 0.2 and 0.4 mg/kg, did not alter the peak area for water. Cocaine at 0.2 mg/kg induced small and reversible increases in choline-containing compounds and N-acetylaspartate peak areas. Cocaine at 0.4 mg/kg induced larger and more sustained increases in choline containing compounds and N-acetylaspartate peak areas. No changes in either water or metabolite resonances were noted following placebo administration. CONCLUSIONS: These increases in choline-containing compounds and N acetylaspartate peak areas may reflect increases in metabolite T2 relaxation times secondary to osmotic stress and/or increased phospholipid signaling within the basal ganglia following cocaine administration. This is the first report of acute, drug-induced changes in the intensity of human brain proton magnetic resonance spectroscopy resonance areas. PMID- 11032981 TI - Basal plasma dehydroepiandrosterone sulfate level: a possible predictor for response to electroconvulsive therapy in depressed psychotic inpatients. AB - BACKGROUND: Dehydroepiandrosterone (DHEA) and its sulfate derivative DHEAS are neuroactive steroids. In the brain, they interact with gamma-aminobutyric acid (GABA(A)) receptors, which are involved in the regulation of anxiety and mood. The relevance of circulatory neurosteroids to psychiatric disorders and biological treatment is unknown. METHODS: Basal plasma levels of cortisol, DHEA, and DHEAS and the DHEAS-DHEA ratio were determined in 17 psychiatric inpatients before and after six electroconvulsive (ECT) therapy sessions, and all changes were statistically analyzed. For baseline values, 25 healthy individuals served as control subjects. Severity of depression and psychosis in the patients was measured with the Hamilton Depression Rating Scale (HDRS) and the Brief Psychiatric Rating Scale, respectively. RESULTS: Both basal and post-ECT levels of cortisol, DHEA, and DHEAS were significantly higher in the patients than in the control subjects. DHEAS levels in responding patients were higher at completion of treatment than at baseline. Patients defined as ECT nonresponders (change in HDRS < 30% from before treatments) exhibited elevated basal DHEAS levels compared with ECT responders. CONCLUSIONS: Markedly elevated basal DHEAS levels (mean + 2 SD of control value) are associated with resistance to ECT and may serve as a potential predictive marker of nonresponsiveness to ECT in depressed patients. PMID- 11032982 TI - Lateralized P3 deficit in schizotypal personality disorder. AB - BACKGROUND: Reduced, left-lateralized P3 amplitude has been reported in several studies focusing on electrophysiologic function in schizophrenia. Also, several lines of evidence suggest a similarity between schizophrenia and schizotypal personality disorder (SPD). This study was undertaken to determine the replicability of our previous finding of a left-lateralized P3 amplitude deficit in SPD. METHODS: We recorded event-related potentials in 21 SPD and 18 normal control subjects in an auditory "oddball" P3 paradigm. RESULTS: In the SPD subjects, but not in the control subjects, there was lower P3 amplitude at T3 compared with T4. CONCLUSIONS: These results are similar to the ones in our previous work and further support the presence of a left-lateralized P3 deficit in SPD. PMID- 11032983 TI - No evidence for a parent-of-origin effect detected in the pattern of inheritance of schizophrenia. AB - BACKGROUND: Schizophrenia is a complex genetic disorder with no clear pattern of inheritance. Epigenetic modification of genes may thus play a role in its transmission. METHODS: In our study, 439 families with at least two ill siblings with schizophrenia (208 with unilineal transmission) were examined for evidence of a parent-of-origin effect (e.g., evidence of parental imprinting on the familial transmission of schizophrenia). RESULTS: No significant difference in the prevalence of maternal compared with paternal transmission was found. In addition, affected male subjects did not differ from affected female subjects in the proportion of their offspring diagnosed with schizophrenia. CONCLUSIONS: Although the transmission of schizophrenia may be influenced by epigenetic events, our study fails to find evidence that one epigenetic mechanism, a parent of-origin imprinting effect, determines whether an individual expresses the illness. PMID- 11032984 TI - Expression and conditioned inhibition of fear-potentiated startle after stimulation and blockade of AMPA/Kainate and GABA(A) receptors in the dorsal periaqueductal gray. AB - Previous work showed that the dorsal periaqueductal gray is involved in the inhibition of fear-potentiated startle. The present study investigated the effects of blockade and stimulation of Kainate/AMPA and GABA(A) receptors within the dorsal periaqueductal gray on expression and conditioned inhibition of fear potentiated startle. Blockade of the Kainate/AMPA receptors enhanced whereas stimulation of the Kainate/AMPA receptors decreased expression of fear potentiated startle. These effects do not reflect conditioned inhibition since this modulation was not changed by injections of Kainate/AMPA receptor agonists or antagonists into the dorsal periaqueductal gray. Stimulation and blockade of GABA(A) receptors within the dorsal periaqueductal gray neither affected expression of fear-potentiated startle nor its conditioned inhibition. The present results together with findings from the literature indicate that glutamate in the dorsal periaqueductal gray is a critical substrate for the expression and modulation of fear-related behaviours. PMID- 11032985 TI - Autoradiographic localization of [3H]nociceptin binding sites in the rat brain. AB - The binding sites of nociceptin (also named orphanin FQ), the endogenous ligand of ORL1 (opiate receptor like 1), were localized in rat brain, using an autoradiographic procedure. High levels of binding were observed in the cingulate, retrosplenial, perirhinal, insular and occipital cortex, anterior and posteromedial cortical amygdaloid nuclei, basolateral amygdaloid nucleus, amygdaloid complex, posterior hippocampus, dorsal endopiriform, central medial thalamic, paraventricular, rhomboid thalamic, suprachiasmatic, ventromedial hypothalamic nuclei, mammillary complex, superficial gray layer of the superior colliculus, locus coeruleus, dorsal raphe nucleus. More moderate labelling was observed in the prefrontal, fronto-parietal, temporal, piriform cortex, dentate gyrus, anterior olfactory nucleus, olfactory tubercle, shell of nucleus accumbens, claustrum, lateral septum, laterodorsal thalamic, medial habenular, subthalamic, reuniens thalamic nuclei, subiculum, periaqueductal grey matter and pons. A lower binding site density was observed in the anterior and medial hippocampus, olfactory bulb, caudate putamen, the core of the nucleus accumbens, medial septum, ventrolateral, ventroposterolateral and mediodorsal thalamic nuclei, lateral and medial geniculate nuclei, hypothalamic area, substantia nigra, ventral tegmentum area and interpedoncular nucleus. A moderate and similar labelling was found in the dorsal and ventral horn of the spinal cord. No labelling was apparent in the corpus callosum. Thus, it appears that the ORL1 receptor is particularly abundant in the cerebral cortex, limbic system of the rat brain and some areas involved in pain perception. PMID- 11032986 TI - Antihistamine terfenadine potentiates NMDA receptor-mediated calcium influx, oxygen radical formation, and neuronal death. AB - We previously reported that the histamine H1 receptor antagonist terfenadine enhances the excitotoxic response to N-methyl-D-aspartate (NMDA) receptor agonists in cerebellar neurons. Here we investigated whether this unexpected action of terfenadine relates to its antihistamine activity, and which specific events in the signal cascade coupled to NMDA receptors are affected by terfenadine. Low concentrations of NMDA (100 microM) or glutamate (15 microM) that were only slightly (<20%) toxic when added alone, caused extensive cell death in cultures pre-exposed to terfenadine (5 microM) for 5 h. Terfenadine potentiation of NMDA receptor response was mimicked by other H1 antagonists, including chlorpheniramine (25 microM), oxatomide (20 microM), and triprolidine (50 microM), was prevented by histamine (1 mM), and did not require RNA synthesis. Terfenadine increased NMDA-mediated intracellular calcium and cGMP synthesis by approximately 2.4 and 4 fold respectively. NMDA receptor-induced cell death in terfenadine-treated neurons was associated with a massive production of hydrogen peroxides, and was significantly inhibited by the application of either (+)-alpha-tocopherol (200 microM) or the endogenous antioxidant melatonin (200 microM) 15 min before or up to 30 min after receptor stimulation. This operational time window suggests that an enduring production of reactive oxygen species is critical for terfenadine-induced NMDA receptor mediated neurodegeneration, and strengthens the importance of antioxidants for the treatment of excitotoxic injury. Our results also provide direct evidence for antihistamine drugs enhancing the transduction signaling activated by NMDA receptors in cerebellar neurons. PMID- 11032987 TI - Changes of GABA(A) receptor binding and subunit mRNA level in rat brain by infusion of subtoxic dose of MK-801. AB - In the present study, we have investigated the effects of prolonged inhibition of NMDA receptor by infusion of subtoxic dose of MK-801 to examine the modulation of GABA(A) receptor binding and GABA(A) receptor subunit mRNA level in rat brain. It has been reported that NMDA-selective glutamate receptor stimulation alters GABA(A) receptor pharmacology in cerebellar granule neurons in vitro by altering the levels of selective subunit. However, we have investigated the effect of NMDA antagonist, MK-801, on GABA(A) receptor binding characteristics in discrete brain regions by using autoradiographic and in situ hybridization techniques. The GABA(A) receptor bindings were analyzed by quantitative autoradiography using [3H]muscimol, [3H]flunitrazepam, and [35S]TBPS in rat brain slices. Rats were infused with MK-801 (1 pmol/10 microl per h, i.c.v.) for 7 days, through pre implanted cannula by osmotic minipumps (Alzet, model 2 ML). The levels of [3H]muscimol binding were highly elevated in almost all of brain regions including cortex, caudate putamen, thalamus, hippocampus, and cerebellum. However, the [3H]flunitrazepam binding and [35S]TBPS binding were increased only in specific regions; the former level was increased in parts of the cortex, thalamus, and hippocampus, while the latter binding sites were only slightly elevated in parts of thalamus. The levels of beta2-subunit were elevated in the frontal cortex, thalamus, hippocampus, brainstem, and cerebellar granule layers while the levels of beta3-subunit were significantly decreased in the cortex, hippocampus, and cerebellar granule layers in MK-801-infused rats. The levels of alpha6- and delta-subunits, which are highly localized in the cerebellum, were increased in the cerebellar granule layer after MK-801 treatment. These results show that the prolonged suppression of NMDA receptor function by MK-801-infusion strongly elevates [3H]muscimol binding throughout the brain, increases regional [3H]flunitrazepam and [35S]TBPS binding, and alters GABA(A) receptor subunit mRNA levels in different directions. The chronic MK-801 treatment has differential effect on various GABA(A) receptor subunits, which suggests involvement of differential regulatory mechanisms in interaction of NMDA receptor with the GABA receptors. PMID- 11032988 TI - Distinctive amygdala kindled seizures differentially affect neurobehavioral recovery and lesion-induced basic fibroblast growth factor (bFGF) expression. AB - The differing effects of partial seizures on neurobehavioral recovery following anteromedial cortex (AMC) injury in rats have previously been reported. Specifically, convulsive Stage 1 seizures evoked ipsilateral to the lesion during the 6-day post-lesion critical period delayed recovery, while non-convulsive Stage 0 seizures were neutral. The present study was designed to elaborate on that research by examining several potential mechanisms for the seizure associated difference observed in functional outcome. Anesthetized rats sustained unilateral AMC lesions followed by implantation of a stimulating electrode in the amygdala ipsilateral (Expt. 1) or contralateral (Expt. 2) to the lesion. Beginning 48 h after surgery, animals were kindled to evoke Stage 0 or Stage 1 seizure activity during the critical period. Kindling trials and afterdischarge (AD) were controlled to ascertain their role in functional outcome. Recovery from somatosensory deficits was assessed over a two-month period. The results revealed that (i) Stage 0 seizures did not impact recovery regardless of whether initiated ipsilateral or contralateral to the lesion, (ii) Stage 1 seizures prevented recovery only when initiated in the ipsilateral hemisphere during the post-lesion critical period, and (iii) the detrimental effect of Stage 1 seizures appears to be independent of the number of kindling trials provided and cumulative AD. Thus, to determine why Stage 1 seizures evoked in the hemisphere ipsilateral to the lesion impeded recovery, a separate group of animals (Expt. 3) were kindled accordingly and processed for c-Fos and basic fibroblast growth factor (bFGF) immunohistochemistry. It was hypothesized that Stage 1 seizures evoked in the injured hemisphere prevent recovery by blocking lesion-induced bFGF expression in structures shown to be important for recovery after cortex lesions (e.g., striatum). The results confirmed our hypothesis and suggest that the seizure associated inhibition of lesion-induced bFGF may alter the growth factor-mediated plasticity necessary for functional recovery. PMID- 11032989 TI - Differential modulation of auditory thalamocortical and intracortical synaptic transmission by cholinergic agonist. AB - To investigate synaptic mechanisms underlying information processing in auditory cortex, we examined cholinergic modulation of synaptic transmission in a novel slice preparation containing thalamocortical and intracortical inputs to mouse auditory cortex. Extracellular and intracellular recordings were made in cortical layer IV while alternately stimulating thalamocortical afferents (via medial geniculate or downstream subcortical stimulation) and intracortical afferents. Either subcortical or intracortical stimulation elicited a fast, 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX)-sensitive, monosynaptic EPSP followed by long duration, polysynaptic activity. The cholinergic agonist carbachol suppressed each of the synaptic potentials to different degrees. At low concentrations (5 microM) carbachol strongly reduced (>60%) the polysynaptic slow potentials for both pathways but did not affect the monosynaptic fast potentials. At higher doses (10-50 microM), carbachol also reduced the fast potentials, but reduced the intracortically-elicited fast potential significantly more than the thalamocortically-elicited fast potential, which at times was actually enhanced. Atropine (0.5 microM) blocked the effects of carbachol, indicating muscarinic receptor involvement. We conclude that muscarinic modulation can strongly suppress intracortical synaptic activity while exerting less suppression, or actually enhancing, thalamocortical inputs. Such differential actions imply that auditory information processing may favor sensory information relayed through the thalamus over ongoing cortical activity during periods of increased acetylcholine (ACh) release. PMID- 11032990 TI - Inhibitory effect of cyclic guanosine 3',5'-monophosphate (cGMP) on the afferent resting activity in the cephalopod statocyst. AB - The effects of exo- and endogenous cGMP on the resting activity (RA) of afferent crista fibers were studied in isolated preparations of the statocysts of the cuttlefish Sepia officinalis and the squid Sepioteuthis lessoniana. Bath application of the membrane-permeable cGMP analogs 8-bromo-cGMP (B-cGMP) and N(2),2'-o-dibutyryl 3', 5'-cyclic guanosine monophosphate (dB-cGMP), and of the selective inhibitor of cGMP-phosphodiesterase zaprinast (ZAP), caused an inhibition of RA. The inhibitory effects of B-cGMP and dB-cGMP remained when the preparation was pre-treated with: (i) the guanylate cyclase inhibitors 1H [1,2,4]oxadiazolo[4,3, -a]quinoxalin-1-one (ODQ) or cystamine (CYS); (ii) the adenylate cyclase inhibitors nicotinic acid (NIC-A), 2',3'dideoxyadenosine (DDA), or MDL-12330A (MDL); (iii) the guanylate cyclase inhibitor methylene blue (M-BLU) and the adenylate cyclase inhibitor MDL combined; or (iv) the nitric oxide (NO) synthase inhibitors N(G)-nitric-L-arginine methyl ester HCl (L-NAME) or N(G) nitro-L-arginine (L-NOARG). These data indicate that cGMP, as an intracellular messenger, has a tonic inhibitory effect on the RA of afferent crista fibers in cephalopod statocysts. PMID- 11032991 TI - Spatial distribution of semicircular canal nerve evoked monosynaptic response components in frog vestibular nuclei. AB - Most second-order vestibular neurons receive a canal-specific monosynaptic excitation, although the central projections of semicircular canal afferents overlap extensively. This remarkable canal specificity prompted us to study the spatial organization of evoked field potentials following selective stimulation of individual canal nerves. Electrically evoked responses in the vestibular nuclei were mapped systematically in vitro. Constructed activation maps were superimposed on a cytoarchitectonically defined anatomical map. The spatial activation maps for pre- and postsynaptic response components evoked by stimulation of a given canal nerve were similar. Activation maps for monosynaptic inputs from different canals tended to show a differential distribution of their peak amplitudes, although the overlap was considerable. Anterior vertical canal signals peaked in the superior vestibular nucleus, posterior vertical canal signals peaked in the descending and in the dorsal part of the lateral vestibular nucleus, whereas horizontal canal signals peaked in the descending and in the ventral part of the lateral vestibular nucleus. A similar, differential but overlapping, spatial organization of the canal inputs was described also for other vertebrates, suggesting a crude but rather conservative topographical organization of semicircular canal nerve projections within the vestibular nuclei. Differences in the precision of topological representations between vestibular and other sensory modalities are discussed. PMID- 11032992 TI - Chronic lithium and sodium valproate both decrease the concentration of myo inositol and increase the concentration of inositol monophosphates in rat brain. AB - One of the mechanisms underlying lithium's efficacy as a mood stabilizer in bipolar disorder has been proposed to be via its effects on the phosphoinositol cycle (PI-cycle), where it is an inhibitor of the enzyme converting inositol monophosphates to myo-inositol. In contrast, sodium valproate, another commonly used mood stabilizer, appears to have no direct effects on this enzyme and was thus believed to have a different mechanism of action. In the present study, high resolution nuclear magnetic resonance (NMR) spectroscopy was used to study the chronic effects of both lithium and sodium valproate on the concentrations of myo inositol and inositol monophosphates in rat brain. As predicted, lithium-treated rats exhibited a significant increase in the concentration of inositol monophosphates and a significant decrease in myo-inositol concentration compared to saline-treated controls. However, unexpectedly, sodium valproate administration produced exactly the same results as lithium administration. These novel findings suggest that both lithium and sodium valproate may share a common mechanism of action in the treatment of bipolar disorder via actions on the PI cycle. PMID- 11032993 TI - Estrogen receptors and aromatase activity in the hypothalamus of the female frog, Rana esculenta. Fluctuations throughout the reproductive cycle. AB - It is well known that certain actions of androgen are mediated through in situ aromatization to estrogen in neural target tissues. This study was undertaken to investigate androgen utilization in the hypothalamus of the female frog, Rana esculenta, through a quantification of estrogen receptors and aromatase activity during the reproductive cycle. 3H-estradiol-binding molecules were present in both the cytosol and the nuclear extract of the hypothalamus. These molecules bound specifically 3H-estradiol with high affinity (Kd 10(-10) M) and low capacity (cytosol: 1.2+/-0.4 fmol/mg protein; nuclear extract: 7.9+/-0.6 fmol/mg protein). Aromatase activity was detected in the microsomal fraction of the hypothalamus using a sensitive in vitro radiometric assay. Both aromatase activity and nuclear estrogen receptor binding fluctuated in synchrony throughout the reproductive cycle. Western blot analysis of aromatase protein revealed one immunoreactive band with a molecular weight of approximately 56 kDa. In contrast to aromatase enzyme activity, the relative levels of aromatase protein changed little during the reproductive cycle suggesting that post-translational mechanisms may be involved in regulating estrogen synthesis in the frog brain. A possible role for estrogens in the modulation of the reproductive behavior in this species is suggested. PMID- 11032994 TI - Improgan, a cimetidine analog, induces morphine-like antinociception in opioid receptor-knockout mice. AB - Improgan is an analog of the H(2) antagonist cimetidine that does not act on known histamine receptors, but induces highly effective analgesia in rodents following intracerebroventricular (icv) administration. Since the mechanism of action of this compound remains unknown, improgan analgesia was characterized presently with the tail immersion nociceptive test in mutant mice lacking either the mu (exon 1 of MOR-1), delta (exon 2 of DOR-1) or kappa (exon 3 of KOR-1) opioid receptor. Improgan (30 microg, icv) induced reversible, maximal analgesia in both sexes of all three genotypes (+/+, +/- and -/-) of MOR-1 mutant mice 10 and 20 min after administration, whereas morphine analgesia was reduced (+/-) or abolished (-/-) in these subjects. In DOR-1 mutant mice, improgan was equally effective in all three genotypes, despite the reduction (+/-) or complete loss ( /-) of delta opioid receptor (3H-[D-Pen(2), D-Pen(5)]enkephalin, DPDPE) binding. Similarly, improgan analgesia was equivalent in all three genotypes of KOR-1 mutant mice, whereas kappa-mediated analgesia (U50,488) and kappa opioid (3H U69,593) binding were abolished in the homozygous (-/-) mice. These studies demonstrate that improgan analgesia does not require intact MOR-1, DOR-1, or KOR 1 genes, and support the hypothesis that improgan-like analgesics act in the CNS by non-opioid mechanisms. PMID- 11032995 TI - Influence of estrous cycle on vaginocervical sensitivity: a fos immunohistochemical study of lumbosacral spinal cord. AB - Expression of c-fos in L(5)-S(1) spinal segments in response to mechanical vaginocervical stimulation was investigated in both cycling and ovariectomized females. The aim of this paper was to verify the influence of estrous cycle on females genital tract sensitivity using immunodetection of a neural activity endogenous marker. The results indicate that lumbosacral spinal Fos-labeling was highly increased in vaginocervical stimulated rats relative to control, and labeled neurons were present more intensively in the dorsal horn in comparison to other spinal areas. Significant differences in Fos-labeling were observed according to the estrous cycle stage at which the stimulation was applied. In estrous females, the response was greater than that obtained at diestrous and much greater than the response of proestrous females. The spinal Fos-labeling of ovariectomized females is equivalent to that of diestrous females. These results give evidence that the vaginocervical induced expression of c-fos is modulated by cyclic changes in circulating sex hormones, whereas results observed in ovariectomized females indicate the likely involvement of other mechanisms independent of ovarian hormones. PMID- 11032996 TI - Specific subnuclei of the nucleus tractus solitarius play a role in determining the duration of inspiration in the rat. AB - Our previous data obtained in the cat suggest that the neurons of the ventrolateral subnucleus of the tractus solitarius (vlNTS) act as an inspiratory off-switch and terminate the inspiratory phase of the respiratory cycle (Berger et al., Eur. J. Pharmacol. 277 (1995) 195-208; Gillis et al., Neurosci. Abstr. 23 (1997) 725). The purpose of the present study was to determine whether inhibition of the region of the vlNTS of the rat using drugs that hyperpolarize, disfacilitate or block both axonal conduction and action potential generation would alter the inspiratory phase of the respiratory cycle. Experiments were conducted in anesthetized, vagotomized and spontaneously breathing rats while monitoring diaphragmatic electromyogram activity. Vagus nerves were sectioned in order to rule out prolongation of inspiration evoked by microinjection of agents into the vlNTS which block excitatory drive from lung afferent inputs. Bilateral microinjection of the inhibitory amino acid gamma-aminobutyric acid (GABA) 25 nmol/45 nl produced an immediate prolongation of inspiratory duration (484+/-18 to 1291+/-84 ms) and an apneustic pattern of breathing. Other effects observed were a significant shortening of expiratory duration (778+/-36 to 432+/-38 ms), rise in blood pressure (83+/-4 to 108+/-6 mmHg) and a small but significant increase in heart rate (439+/-17 to 452+/-18 beats/min). Bilateral microinjection of the ionotropic glutamate receptor antagonist kynurenic acid (1 nmol) and the Na(+) channel blocker tetrodotoxin (10 pmol) into the region of the vlNTS consistently produced a similar prolongation of inspiratory duration and an apneustic pattern of breathing. These results support the hypothesis that neurons in the region of the vlNTS promote the transition from inspiration to expiration and function as part of the 'Inspiratory Off Switch'. PMID- 11032997 TI - Serotonergic mechanisms of the lateral parabrachial nucleus on DOCA-induced sodium intake. AB - It has been shown that the serotonergic mechanisms of the lateral parabrachial nucleus (LPBN) inhibit NaCl intake in different models of angiotensin II (ANG II) dependent NaCl intake in rats. However, there is no information about the involvement of LPBN serotonergic mechanisms on NaCl intake in a model of NaCl intake not dependent on ANG II like deoxycorticosterone (DOCA)-induced NaCl intake. Therefore, in this study we investigated the effects of bilateral injections of serotonergic agonist and antagonist into the LPBN on DOCA-induced 1.8% NaCl intake in rats. Male Holtzman rats were treated with s.c. DOCA (10 mg/rat each every 3 days). After a period of training, in which the rats had access to 1.8% NaCl during 2 h for several days, the rats were implanted with stainless steel cannulas bilaterally into the LPBN. Bilateral injections of the serotonergic receptor antagonist methysergide (4 microg/0.2 microl each site) in the LPBN increased 1.8% NaCl intake (32.2+/-3.9 versus vehicle: 15.0+/-1.6 ml/2 h, n=10) and water intake (12.5+/-3.5 versus vehicle: 3.2+/-1.0 ml/2 h). Injections of the serotonergic 5HT(2A/2C) receptor agonist DOI (5 microg/0,2 microl each site) in the LPBN reduced 1.8% NaCl intake (6.8+/-1.7 versus saline: 12.4+/-1. 9 ml/2 h, n=10) and water intake (2.2+/-0.8 versus saline: 4.4+/-1.0 ml/2 h). Besides the previously demonstrated importance for the control of ANG II dependent water and NaCl intake, the data show that the serotonergic inhibitory mechanisms of the LPBN are also involved in the control of DOCA-induced NaCl intake. PMID- 11032998 TI - Alteration of mitochondrial calcium homeostasis by ammonia-induced activation of NMDA receptors in rat brain in vivo. AB - The aim of the present work was to assess the effects of activation of NMDA receptors in rat brain in vivo on calcium homeostasis in isolated non-synaptic brain mitochondria. We have shown recently that acute intoxication with large doses of ammonia leads to activation of NMDA receptors in rat brain in vivo. In the present work we injected rats with ammonium acetate to activate NMDA receptors in vivo and isolated non-synaptic mitochondria to assess calcium homeostasis. We also tested whether blocking NMDA receptors with MK-801 prevents effects on calcium homeostasis induced by ammonium injection. It is shown that activation of NMDA receptors in rat brain in vivo leads to a rapid increase in intramitochondrial calcium content followed by a reduction in the calcium capacity and calcium uptake rate in rat brain mitochondria. Activation of NMDA receptors resulted in increased spontaneous calcium efflux from rat brain mitochondria and in a strong inhibition of Na-induced and tert-butylhydroperoxide induced calcium efflux. All these effects were prevented by previous blocking of NMDA receptors by injection of MK-801. Cyclosporin A did not affect any of the above parameters, indicating that the mitochondrial permeability transition pore does not play a role in calcium efflux under any of the conditions studied. The results reported indicate that ammonia-induced activation of NMDA receptors in rat brain in vivo alters mitochondrial calcium homeostasis at several different steps. PMID- 11033000 TI - The effects of potassium on the rat middle cerebral artery. AB - After traumatic brain injury, extracellular K(+) in brain can dramatically increase. We studied the effects of increased K(+) on the isolated pressurized rat middle cerebral artery (MCA). MCAs (200-250 microm OD) were isolated, cannulated with glass micropipettes, and pressurized. K(+) was increased in the extraluminal bath using three paradigms: (1) isotonic K(+) (K(iso)) where increases in K(+) were offset by decreases in Na(+), (2) hypertonic K(+) (K(hyper)) where K(+) was increased without a concomitant adjustment of Na(+), and (3) K(suc), a solution using K(iso) but with the addition of sucrose to obtain a hypertonic solution. Increases in K(+) in the extraluminal bath produced significant dilations (approximately 20%) at 21 mM K(+) in all three groups (K(iso), K(hyper), and K(suc)). With the K(hyper) and K(suc) groups, the magnitude of the dilation diminished with further increases in K(+). L-NAME (10( 5) M), an inhibitor of nitric oxide synthase, had no effect on the response of the K(hyper) and K(suc) groups at 21 mM but significantly enhanced constrictions of the MCAs above 40 mM K(+) compared to the control. The K(iso) group was not affected by L-NAME at any K(+) concentration and showed profound constrictions above 40 mM K(+). We conclude that changes in the K(+) concentration and osmolality of the extracellular fluid may have profound effects on the cerebral vasculature. PMID- 11032999 TI - Regulation of alpha(2A)-adrenoceptor expression by chronic stress in neurons of the brain stem. AB - Alpha(2)-Adrenoceptors are supposed to be important regulatory elements in responses to stress. Previous receptor binding studies in male tree shrews have shown that chronic psychosocial stress down-regulates binding sites for alpha(2) adrenergic ligands in several brain stem nuclei. The aim of the present study was to quantify effects of chronic subordination stress on expression of the alpha(2) adrenoceptor subtype A gene in identified neurons of the brain stem. We partially cloned the alpha(2A)-adrenoceptor cDNA of the tree shrew (1.22 kb) and localized receptor RNA expression in brain stem neurons by in situ hybridization using a 35S-labeled cRNA probe (1.06 kb). To identify neurons expressing receptor mRNA, brain sections were first immunocytochemically stained with antibodies against tyrosine hydroxylase, phenylethanolamine-N-methyltransferase, or glutamate, and then processed for in situ hybridization. Furthermore, expression of receptor specific RNA was quantified in single neurons of animals which had been psychosocially stressed during 4 weeks and in unstressed controls. We found strong in situ hybridization in the noradrenergic neurons of the locus coeruleus, but only weak labeling of A2 neurons in the solitary tract nucleus and no labeling of A1 neurons in the caudal ventrolateral medulla. Adrenergic neurons in the solitary tract nucleus (group C2) did not express the alpha(2A)-adrenoceptor, and C1 neurons in the rostral ventrolateral medulla showed only a minor labeling by the in situ probe. In contrast, large glutamatergic neurons in the lateral reticular nucleus were strongly labeled by the probe. Chronic psychosocial stress reduced alpha(2A)-adrenoceptor RNA expression in locus coeruleus neurons ( 24.0%), in solitary tract neurons (-31.0%), and in neurons of the lateral reticular nucleus (-18.8%). These findings show that stress not only decreases the expression of the alpha(2A)-adrenergic autoreceptor in the locus coeruleus but also of alpha(2A)-heteroreceptors in glutamatergic neurons. PMID- 11033001 TI - Persistent and delayed behavioral changes after nicotine treatment in adolescent rats. AB - Despite the increasing use of tobacco by adolescents, few animal studies have addressed the neurobehavioral consequences of nicotine exposure during this period. We administered nicotine to adolescent rats via continuous infusion on postnatal days (PN) 30 through 47.5, using a dosage regimen that maintains plasma levels similar to those found in smokers or in users of the transdermal nicotine patch. Behavior in a novel open field and learning a passive avoidance task were assessed during nicotine treatment and for 2 weeks post-treatment. On PN44, during nicotine exposure, female rats showed decreased grooming, an effect not seen in males; this effect is opposite to the effects of nicotine in adult rats. Two weeks after cessation of nicotine administration, females showed deficits in locomotor activity and rearing, whereas males again were unaffected; the behavioral deficits appeared at the same age at which gender-selective brain cell damage emerges. In contrast, nicotine exposure enhanced passive avoidance, with the effect intensifying and persisting throughout the post-treatment period. These results reinforce the concept that developmental vulnerability to nicotine extends into adolescence, with patterns of drug effects different from those in earlier or later periods. The correlation of neurochemical with behavioral effects strengthens the connection between adolescent nicotine exposure and persistent functional changes that may influence drug habituation, learning and memory. PMID- 11033002 TI - Time dependency of the action of nitric oxide in lipopolysaccharide-interferon gamma-induced neuronal cell death in murine primary neuron-glia co-cultures. AB - We investigated the time-dependency of the action of nitric oxide (NO) on glia mediated neuronal cell death. Cortical neuron-glia co-cultures were treated with lipopolysaccharide and interferon gamma (LPS/IFNgamma). The production of NO was first detectable 9 h after the exposure to LPS/IFNgamma and increased for up to 48 h. A significant neuronal cell death was observed 36-48 h after treatment with LPS/IFNgamma. The NO generated at the initial stage of NO synthesis (about 12 h) following exposure to LPS/IFNgamma was found to be critical for LPS/IFNgamma induced neurotoxicity. Furthermore, the rate of NO production at the initial stage of NO synthesis was correlated linearly with the extent of neuronal cell death. These findings suggest that the maximal rate of NO synthesis, instead of the accumulated NO(2)(-) level, is a sensitive index for predicting endotoxin induced cytotoxicity. PMID- 11033003 TI - Stimulation of the MLR inhibits the discharge of dorsal horn neurons responsive to muscular contraction. AB - We found that electrical stimulation of the mesencephalic locomotor region (MLR) inhibited the discharge of deep dorsal horn neurons receiving group III afferent input from the triceps surae muscles. In contrast, contraction of these muscles induced by electrical stimulation of the tibial nerve activated these dorsal horn neurons. Our findings show that descending central motor commands can inhibit dorsal horn interneurons receiving input from group III afferents during exercise. PMID- 11033004 TI - The role of activity blockade on glutamate receptor subunit expression in the spinal cord. AB - Spinal cord injury (SCI) impairs descending glutamatergic neurotransmission reaching ventral motor neurons (VMN). Previously we reported up-regulation of NMDA receptor subunits NR1 and NR2A mRNAs in VMN caudal to the lesion site 24 h after SCI. The absence of effect rostral to the injury site suggested that injury induced loss of descending activity might be involved. To test this hypothesis, we blocked axonal conduction by focal injection of tetrodotoxin into the spinal cord. We found increased NR1 and NR2A mRNAs in VMN similar in extent to that seen after SCI. Thus, the increase in NMDA subunit mRNAs may be 'inactivity dependent' and associated with reorganization of the spinal cord in response to loss of descending innervation. PMID- 11033005 TI - Does fucose or piracetam modify the effect of hypoxia preconditioning against pentylenetetrazol-induced seizures? AB - To clarify the question whether the duration of hypoxia exposure has an influence on the point in time or the strength of hypoxic preconditioning, hypoxia exposure of rats lasting 1 and 8 h was tested regarding the modification of susceptibility to acute pentylenetetrazol-induced seizures. Following the short-lasting (1 h) hypoxia, the maximum level of preconditioning action was observed 7 days after hypoxia, whereas the longer-lasting hypoxia (8 h) produced the maximum level of protection 14 days after hypoxia. We investigated the influence of fucose and piracetam on the effect of hypoxia preconditioning by the application of the substances 20 min before the beginning of hypoxia exposure. Fucose did not modify the result of hypoxia preconditioning. But after the treatment with piracetam, the preconditioning effect was prevented following hypoxia lasting 1 and 8 h. We suggest that the radical scavenger properties of piracetam are responsible for the absence of protection against pentylenetetrazol-evoked seizures. PMID- 11033006 TI - A di-synaptic projection from the lateral cerebellar nucleus to the laterodorsal part of the striatum via the central lateral nucleus of the thalamus in the rat. AB - We have examined a cerebello-thalamo-striatal pathway from the lateral cerebellar nucleus (LCN) to the laterodorsal part of the striatum (LDS) through the central lateral nucleus (CL) using light and electron microscopy through the employment of a combination of anterograde and retrograde tracing techniques. Biotinylated dextran amine (BDA) was injected into the unilateral LCN, and used as an anterograde tracer. Cholera toxin B subunit (CTb), used for light microscopy, and wheat germ agglutinin-horseradish peroxidase (WGA-HRP), used for electron microscopy, were injected into the contralateral LDS as retrograde tracers. Light microscopic analysis showed a good overlap of the distribution of BDA-labeled axon terminals and CTb-labeled neurons in the middle third of the CL in both dorsoventral and rostrocaudal axes on the LDS injection side. Electron microscopy confirmed the presence of direct synaptic contacts between BDA-labeled terminals and WGA-HRP-labeled dendrites in the CL. PMID- 11033007 TI - Tetanus-induced asynchronous GABA release in cultured hippocampal neurons. AB - Asynchronous GABA release was studied in cultured hippocampal neurons using paired whole-cell recordings. Tetanization of the presynaptic GABAergic neuron was accompanied by a train of IPSCs which showed tetanic depression. Asynchronous IPSCs (asIPSCs) also developed during the train and continued for 1.85+/-0.3 s after the stimulation. The threshold frequency for evoking asIPSCs was 10 Hz, while maximal asynchronous activity was achieved at 40 Hz. Perfusion with EGTA-AM blocked asIPSCs. The elevation of [Ca(2+)](i) that accompanies presynaptic action potential firing triggers asynchronous release of GABA vesicles, thereby counteracting tetanic depression of synchronous IPSCs. PMID- 11033008 TI - Enhancement of ethanol reward by dopamine D3 receptor blockade. AB - This experiment examined the influence of U-99194A, a dopamine D3 receptor antagonist, on ethanol's rewarding effect in a place conditioning paradigm. Swiss Webster mice received six pairings of a tactile stimulus with ethanol (2 g/kg, i.p.), U-99194A (20 mg/kg, i. p.) with ethanol, or U-99194A alone. A different stimulus was paired with saline. During conditioning, ethanol or ethanol/U-99194A produced similar increases in locomotor activity. U-99194A alone produced modest increases in activity on some trials. As expected, the 2 g/kg ethanol dose produced a nonsignificant trend towards conditioned place preference. However, U 99194A enhanced the acquisition of ethanol preference, whereas U-99194A alone did not produce either place preference or aversion. The results are consistent with the notion that dopamine D3 systems are important in the response to ethanol and further suggest that D3 receptor blockade increases ethanol reward. PMID- 11033009 TI - A melatonin agonist facilitates circadian resynchronization in old hamsters after abrupt shifts in the light-dark cycle. AB - Age-related changes in the mammalian circadian system may be associated with a decline in circulating melatonin levels. Using 'jet lag' paradigms involving abrupt shifts in the light-dark cycle, we showed that a melatonin agonist, S 20098, accelerated by approximately 25% resynchronization of the circadian activity rhythm in old hamsters to the new light-dark cycle. It suggests the usefulness of melatonin-related compounds to treat circadian disorders associated with aging. PMID- 11033010 TI - L-type calcium channels modulate glutamate-driven bursting activity in the nucleus accumbens in vivo. AB - The majority of adult nucleus accumbens medium spiny neurons exhibit a bistable membrane potential that fluctuates between a relatively hyperpolarized (Down) state (average=-76 mV) and a less hyperpolarized (Up) state (average=-60 mV) near firing threshold. During in vivo extracellular recordings from nucleus accumbens neurons, we used microiontophoresis to apply glutamate and selected neurons that fired in bursting patterns reflecting a subthreshold bistable membrane potential. The average frequency of bursts events was 0.85 Hz. The average burst duration was 392+/-3.5 ms, with an average of 13.4 spikes and an average spike frequency of 30.6+/-3.1 Hz per burst. To determine the involvement of the L-type calcium channel in the bursting pattern, we applied the benzothiazepine L-type calcium channel blocker, diltiazem. Diltiazem rapidly (<2 min) and reversibly decreased the burst duration by 29% and the frequency of spikes within a burst by 30% without changing the overall burst event frequency. The results provide the first in vivo electrophysiological evidence implicating an L-type calcium channel that modulates glutamate-induced burst firing of nucleus accumbens neurons. PMID- 11033011 TI - Changes in estrogen-alpha receptor immunoreactivity during the estrous cycle in lactating dairy cattle. AB - Estradiol is one of the most important hormones in the regulation of estrous behavior, which is at a very low level of expression in the modern dairy cow. In the present study the neuroanatomical distribution of estrogen receptors of the alpha-subtype (ER-alpha) in the bovine hypothalamic area is determined with immunocytochemical methods, at various stages of the estrous cycle. During the luteal phase of the cycle, ER-alpha immunoreactive cells were found in most of the nuclei that are known to express ER-alpha immunoreactivity in other species, like the Bed nucleus of the Stria terminalis, Medial preoptic area, Ventromedial hypothalamus and Arcuate nucleus. During estrus and metestrus, however, no ER alpha immunoreactive cells could be detected in those areas, except for a few in the caudal Arcuate nucleus. The results from the present study indicate that there is a coherent regulation and timing of physiological and behavioral events around ovulation, in which estradiol and its receptor play a key role. PMID- 11033012 TI - Transporter reversal as a mechanism of glutamate release from the ischemic rat cerebral cortex: studies with DL-threo-beta-benzyloxyaspartate. PMID- 11033013 TI - Single-shot plasmid DNA intrasplenic immunization for the production of monoclonal antibodies. Persistent expression of DNA. AB - Monoclonal antibodies (Mc. Abs.) were generated against a 18-kDa protein from Brucella abortus 48 h and 25 days after a single intrasplenic injection of a DNA plasmid containing the expression vector for the protein. Hybridomas were also obtained from spleens injected 3, 5, and 10 days before fusion. Somatic cell fusion of spleen cells from mice, injected with the plasmid DNA, in saline, with the NS-0 myeloma cell line resulted in Mc. Abs of the IgG and IgM Isotypes. IgG antibodies were of the IgG2b and IgG1 subtype. Hybridoma tissue culture supernatants were strongly positive by ELISA at dilutions of up to 1/1200 and produced intense specific bands in immunoblotting. All these antibodies recognized the native recombinant protein (the screening antigen) and some of them also recognized the heat-denatured recombinant 18-kDa protein. When compared to standard procedures of immunization, as well as to intramuscular or gene gun DNA immunizations, this technique results in very early, time saving, strong Mc Abs. It is common knowledge that in order to generate specific hybridomas; spleen cells from immunized animals have to be fused no later than 5 days after the last boost. The fact that through single-shot intrasplenic immunization (SSI) specific hybridomas are generated 25 days after one single injection indicates that the gene coding the p18 protein is being expressed in the spleen for at least 20 days. We propose that plasmid DNA intrasplenic immunization can be a helpful tool for the production of specific hybridomas. This route of immunization could also be helpful in the further understanding of early events of the immune response to genetic immunization by naked DNA injection. PMID- 11033014 TI - IgG antibody levels to meningococcal porins in patient sera: comparison of immunoblotting and ELISA measurements. AB - IgG antibody levels to the meningococcal PorA and PorB proteins in 56 acute and convalescent phase sera from 25 patients with meningococcal disease were compared by immunoblotting and ELISA. Heat-treated outer membrane vesicles from strain 44/76 (B:15:P1.7, 16) served as antigens for immunoblotting, whereas purified P1.7,16 PorA and P15 PorB from the same strain were used as antigens in the ELISA. In the blotting method, IgG binding to the porins was determined by digital scanning of the immunoreactive bands and calculated relative to the PorA binding of a reference serum on each blot. The coefficient of variation for the reference serum was 21.6% (a total of 144 strips) with smaller variations for each day's experiments. Blotting of all 56 sera at the standard 1/200 dilution measured anti-PorA and anti-PorB levels that correlated with those obtained by ELISA (Spearman rank-order correlation coefficient r(s)=0.48; P<0.001). At this dilution, the anti-PorA (r(s)=0.52; P<0. 004) and anti-PorB (r(s)=0.60; P<0.001) levels of the convalescent phase sera (n=29) corresponded with the ELISA measurements, whereas no correlation was found with the results for the acute phase sera, which mostly had low ELISA antibody levels (<2 microg/ml IgG). A corresponding blot analysis of convalescent sera from the seven patients, who had received the 44/76 outer membrane vesicle vaccine, demonstrated a high correlation coefficient for the anti-PorA levels (r(s)=0.95; P<0.001) vs. the ELISA results. No such correlation was observed for the PorB response in these sera, being nine-fold higher than the PorA response, because of a prozone effect on the blots at the standard dilution. However, blotting at a higher serum dilution (1/2000) resulted in anti-PorB levels that also correlated strongly (r(s)=0.93 P<0.001) with the ELISA measurements. PMID- 11033015 TI - Critical residues of epitopes recognized by several anti-p53 monoclonal antibodies correspond to key residues of p53 involved in interactions with the mdm2 protein. AB - The aim of this work was to study the reactivity of antibodies directed against the N-terminus of p53 protein. First, we analysed the cross-reactivity of anti p53 antibodies from human, mouse and rabbit sera with peptides derived from human, mouse and Xenopus p53. Next, we characterized more precisely a series of monoclonal antibodies directed against the N-terminal part of p53 and produced by immunizing mice with either full length human or Xenopus p53. For each of these mAbs we localized the epitope recognized on human p53 by the Spot method of multiple peptide synthesis, defined critical residues on p53 involved in the interaction by alanine scanning replacement experiments and determined kinetic parameters using real-time interaction analysis. These antibodies could be divided into two groups according to their epitopic and kinetic characteristics and their cross-reactivity with murine p53. Our results indicate that critical residues involved in the interaction of some of these mAbs with p53 correspond to key residues on p53 involved in its interaction with the mdm2 protein. These antibodies could, therefore, represent powerful tools for the study of p53 regulation. PMID- 11033016 TI - Flow cytometric analysis of intracellular IFN-gamma, IL-4 and IL-10 in CD3(+)4(+) T-cells from rat spleen. AB - The application of multi-parameter flow cytometry for the assessment of T-cell and cytokine functioning has been used by several groups for studying human and mouse samples, although little has been reported for the rat. Here we report the optimisation of immunofluorescent staining for cell surface and intracellular antigens using three-colour flow cytometric analysis to measure the frequency of rat CD3(+)4(+) T-cells that produce IFN-gamma, IL-4 and IL-10. In vitro stimulation of IFN-gamma production required incubation of splenocytes with PMA and ionomycin in the presence of the protein transport inhibitor brefeldin A for 6 h. Three stimulation protocols for IL-4 and IL-10 production were evaluated. In vitro priming of splenic T-cells with antibodies against CD3 and CD28 and recombinant cytokines (IL-2 and IL-4) for 5 days followed by restimulation with PMA and ionomycin was required to stimulate cells to produce either IL-4 or IL 10. Brefeldin A was found to be a more suitable protein transport inhibitor than monensin. This method will be useful for analysing the nature of individual rat cytokine-producing cells in a variety of experimental model systems. PMID- 11033017 TI - Long PCR detection of the C4A null allele in B8-C4AQ0-C4B1-DR3. AB - The genes coding for the two components of complement 4 (C4), C4A and C4B, are located within the major histocompatibility complex (MHC) on the short arm of chromosome 6. Several studies have shown that deficiency of C4A is associated with systemic lupus erythematosus (SLE), rheumatoid arthritis and scleroderma. A large deletion covering most of the C4A gene and the 21-hydroxylase-A (21-OHA) pseudogene found on the extended haplotype B8-C4AQ0-C4B1-DR3 is estimated to account for approximately two-thirds of C4A deficiency in Caucasian SLE patients. Detection of this C4A null allele has been technically difficult due to the high degree of homology between C4A and C4B, with protein analysis and restriction fragment length polymorphism (RFLP) analysis using Southern blotting being the only approaches available. In this study, a long PCR strategy was used to rapidly genotype for the C4A deletion through specific primer design. The methodology makes use of the unique sequence of the G11 gene upstream of C4A and the sequence of a 6.4 kb retrotransposon, the human endogenous retrovirus HERV-K(C4), which is present in intron 9 of C4A but absent in the case of the deletion. PMID- 11033018 TI - Colorimetric determination of inhibition of hematopoietic progenitor cells in soft agar. AB - In vitro colony forming unit (CFU) assays have been used to measure the effects of compounds that regulate the growth of hematopoietic progenitor cells. These assays are time consuming and subjective and are therefore not amenable to high throughput of large numbers of compounds. Here we have shown that the traditional murine bone marrow CFU assay can be modified into a robust non-subjective colorimetric assay format. 3-[4,5-Dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) was added after colony formation in an agar based 96-well plate culture system. Optical density correlated with increasing cell input concentrations in the presence of growth factor. The linearity of this response was equivalent to the standard CFU assay. Several hematopoietic inhibitors were tested in both assays. Effects on colony number and size were compared to optical density. Compounds that reduced colony numbers with little effect on colony size had identical IC(50) values in both the colorimetric assay and CFU assay. The IC(50) values of compounds that also decreased colony size did not correlate in the two assays. These results demonstrate the utility of the colorimetric assay to rapidly screen for compounds that specifically inhibit hematopoietic progenitor cell colony formation in vitro. PMID- 11033019 TI - Identification of HLA-A*03, A*11 and B*07-restricted melanoma-associated peptides that are immunogenic in vivo by vaccine-induced immune response (VIIR) analysis. AB - With the discovery of increasing numbers of tumor antigens, there is a need to rapidly determine whether these antigens and the individual peptides they express are able to stimulate immune responses in vivo and thus, can be used to construct cancer vaccines. In this study we used the method of vaccine-induced immune response (VIIR) analysis to identify multiple immunogenic peptide epitopes derived from several melanoma associated antigens and presented by HLA-A*03, A*11 and B*07. Thirty-one patients with melanoma were immunized to a polyvalent vaccine containing multiple antigens, including MAGE-3, Melan A/MART-1, gp100 and tyrosinase. Their peripheral blood was tested for peptide-specific, vaccine induced CD8+ T cell responses before and after immunization using an enzyme linked immune spot (ELISPOT) assay with panels of peptides restricted by these three alleles. The peptides were selected for immunogenic potential based on their strong binding affinity in vitro to HLA-A*03, A*11 or B*07. Overall, 60% of the 20 peptides studied were recognized by at least one patient and 50% of the patients showed a vaccine-induced CD8+ T cell response to at least one peptide that matched their HLA specificity. We conclude that VIIR analysis is an effective strategy to directly identify immunogenic peptides that are good candidates for vaccine construction. PMID- 11033020 TI - Isolation and serological analyses of fungal melanins. AB - Melanins are notoriously difficult to work with because of their unique physical and chemical properties. The study of melanins is hampered by the scarcity of melanin-specific reagents and serological techniques. In this study we describe modifications to the standard method for the isolation of melanins from in vitro melanized fungal cells and detail the optimization of serological techniques for the study of melanin compounds. The isolation procedure involves the digestion of melanized cells with a combination of proteolytic and glycolytic enzymes, denaturant, organic extractions, and boiling in 6.0 M HCl. Elemental quantitative analyses suggest that this procedure does not significantly affect the relative elemental composition of melanins. For the serological assays, our goal was to achieve a homogenous distribution of melanin particles on a solid support to maximize their recognition by melanin-binding antibodies. The results from enzyme linked immunosorbent assays (ELISAs) demonstrate that melanins, in general, disperse more efficiently on, and adhere better to, medium-binding polystyrene surfaces, especially in the presence of trace amounts of salt. Blocking the melanin-coated ELISA plates with the commercially available SuperBlock((R)) Blocking Buffer for 4 h was more efficient at reducing non-specific binding of a negative control monoclonal antibody (mAb) compared to blocking with 2% bovine serum albumin (BSA) and 5% milk. Increasing the ionic strength of the antibody solutions reduced binding to the melanins, indicating that binding is in part mediated by electrostatic interactions. These conditions were also applied to immunofluorescence (IF) analyses of melanins, and the results were consistent with those obtained by ELISA. PMID- 11033021 TI - Quantitation of antigen-reactive T cells in peripheral blood by IFNgamma-ELISPOT assay and chromium-release assay: a four-centre comparative trial. AB - The ELISPOT assay is increasingly being used for the monitoring of the induction of antigen-reactive T cells in cancer vaccination trials. In order to evaluate the reliability of T cell frequency analysis with the ELISPOT assay, a comparative study was performed in four European laboratories. Six samples from healthy subjects were analyzed for the frequency of influenza-reactive CD8+ T cells in peripheral blood mononuclear cells (PBMC) by IFNgamma-ELISPOT assay. In addition, one laboratory determined cytotoxic T cell precursor (CTL) frequencies in these samples by limiting dilution chromium-release assay (LDA), and three laboratories performed a variant of the LDA, the multiple microculture assay (MMA). Consistent frequencies of influenza peptide-reactive T cells were obtained with the ELISPOT assay in all four laboratories. The numbers detected by ELISPOT assay correlated closely with those determined by LDA. In contrast, the frequencies obtained with the MMA differed considerably and showed little correlation with the other two assays. This study shows that it is possible to use the ELISPOT assay to determine with reliability antigen-reactive T cells in a multicenter setting. We suggest that this assay may be suitable for monitoring cancer vaccine trials. PMID- 11033022 TI - A novel high-throughput method for accurate, rapid, and economical measurement of multiple type 1 diabetes autoantibodies. AB - Prediction of Type 1 diabetes for study of preventive therapies requires screening the general population, where 85% of new cases occur. Even with HLA based prescreening, nearly 20% of all children will need multiple serum autoantibody testings. High-throughput, economical, and accurate methods are therefore essential. We have developed such a radiobinding method, using 96-well microtiter plates and a novel immune complex capture method via membrane-bound Protein A. Each microtiter plate contained a standard negative control serum, and low-, medium-, and high-level positive control sera. All sera were evaluated in triplicate. This readily allowed quality control criteria both for triplicates of individual sera and for each 96-well plate. Inter-assay coefficients of variation (CVs) were all 0 could also be calculated instantly. PMID- 11033027 TI - Quantitative assessment of bleomycin-induced poly(ADP-ribosyl)ation in human lymphocytes by immunofluorescence and image analysis. AB - Poly(ADP-ribose) polymerase (PARP) is a nuclear enzyme that is catalytically activated by DNA strand interruptions. It catalyses the covalent modification of proteins with ADP-ribose polymers, using NAD(+) as precursor. Here, we have studied the DNA damage-induced formation of poly(ADP-ribose) in intact human peripheral blood lymphocytes (PBL) by in-situ immunofluorescence detection. The response of PBL to bleomycin (BLM), which is known to induce DNA single and double strand breaks, was investigated with regard to polymer formation. For this purpose, a quantitative approach was developed to assess more accurately the immunostaining of polymer formation by computerised image analysis. As an application of this new method, we have determined the polymer formation following BLM treatment in quiescent human PBL versus mitogen activated cells. Quiescent human PBL showed a similar basal immunostaining for the polymer compared to phytohemagglutinin (PHA)-activated cells, expressed as relative mean pixel intensity (RMPI) (1.3+/-0.8 and 2.2+/-0.9, respectively; P<0.3). After BLM treatment, there was a clear-cut enhancement of polymer immunostaining, with PHA activated cells showing significantly higher RMPI than non-activated cells (9.2+/ 1.4 and 4.2+/-1.0, respectively; P<0.005). As expected, in the presence of the ADP-ribosylation inhibitor 3-aminobenzamide (3-AB), the RMPI of immunostained polymer was decreased in both quiescent and PHA-activated PBL to 1.2+/-0.7 and 1.5+/-0.9, respectively. Our findings reveal (i) that mitogen-stimulated, intact lymphocytes show enhanced polymer formation following BLM treatment, and (ii) that our new quantitative immunofluorescence assay coupled with computerised image analysis is reliable and sensitive enough to detect changes in polymer formation rate. PMID- 11033026 TI - Apoptosis: a method for evaluating the cryopreservation of whole blood and peripheral blood mononuclear cells. AB - We sought to compare the effect of cryopreservation and storage at -30 degrees C, -70 degrees C and -150 degrees C of human whole blood versus matched peripheral blood mononuclear cell (PBMC) samples using apoptosis as an indicator of cell fitness. Following 10 weeks of storage the samples were thawed and assessed for viability (trypan blue exclusion), levels of apoptosis (using the nuclear stain bis-benzimide) and cell function (ability to be transformed by Epstein-Barr virus, EBV). When comparing storage temperatures, the levels of apoptosis in whole blood and PBMC samples stored at -30 degrees C were significantly higher than the values for samples stored at -70 degrees C or -150 degrees C (P<0.004). Whole blood samples stored at -150 degrees C had significantly less apoptosis than those stored at -70 degrees C (P<0.03). A comparison of the cell preparations showed that at all three storage temperatures there was significant sample deterioration (viability, apoptosis, and function) in whole blood relative to PBMC samples. This study indicates that careful consideration should be given to storage conditions and that apoptosis can be used as a sensitive measure of cell fitness following cryopreservation. PMID- 11033028 TI - A rapid, quantitative method for measuring leukocyte adhesion to normal and balloon-injured arteries in vitro. AB - Many of the currently available techniques for quantifying leukocyte adhesion require monolayers of cells and are therefore unsuitable for use in ex vivo arterial tissue. Here we describe a rapid method to measure adhesion of leukocytes to intact artery strips and to determine the effect of artery injury on adhesiveness of leukocytes with and without activation. Leukocytes were isolated from rabbit blood, labelled with 51Cr, and added to the luminal face of the left and right subclavian arteries derived from the same animal. In some experiments the endothelium was removed before addition of leukocytes and in another series of experiments the artery was injured by inflating a balloon catheter within the lumen in vitro before leukocyte addition. After washing, the adhesion of labelled leukocytes was quantified by gamma counting. To determine localization of the leukocytes, some arteries were fixed in situ and examined microscopically, with confirmation of leukocyte identification by enzyme cytochemistry. The adhesion of leukocytes increased progressively during 60 min and was inhibited by reducing the temperature to 4 degrees C. Adhesion was increased by the nitric oxide synthase inhibitor L-NAME. Stretching the artery wall in vitro using a balloon catheter increased leukocyte adhesion within 1 h after injury. In contrast, this did not occur following simple arterial denudation. Histological examination of stained en face preparations and transverse sections of the subclavian arteries revealed loosely adherent granulocytic leukocytes on the endothelial surface. This technique is straightforward and allows accurate and rapid measurement of autologous leukocyte adhesion to normal and pathologically altered arteries ex vivo. PMID- 11033029 TI - Development of highly sensitive enzyme-linked immunosorbent assays for hepatocyte growth factor/scatter factor (HGF/SF): determination of HGF/SF in serum and urine from normal human subjects. AB - A sandwich enzyme-linked immunosorbent assay (ELISA) using rabbit anti-hepatocyte growth factor (HGF) IgG for human HGF, also known as the scatter factor, has previously been developed for determining increases in serum HGF levels in various liver diseases. The sensitivity limit of the ELISA is, however, approximately 0.2 ng/ml sample, and HGF concentrations in about 50% of normal subjects are not accurately measurable by this method, because the mean level of HGF in normal serum is close to the sensitivity limit. In the present study, chicken Fab' from egg yolk anti-HGF immunoglobulin Y and rabbit Fab' from rabbit anti-HGF IgG were conjugated with beta-D-galactosidase. With these conjugates as the second antibodies, we developed two sandwich ELISAs for human HGF and found that the sensitivities were about 20 pg/ml with the former conjugate and 2 pg/ml with the latter. The HGF concentration in sera from 138 normal subjects determined by the ELISA with the rabbit conjugate was 244+/-65 (SD) pg/ml serum, and it correlated very well with the number of leukocytes. Moreover, the ELISA with the rabbit conjugate permitted the determination of HGF levels in urine from normal subjects without first concentrating the sample. The determination of HGF in various biological fluids other than blood and urine by these ELISAs may aid the diagnosis and prognosis of various diseases. PMID- 11033030 TI - Isolation of antibodies which neutralise the activity of early pregnancy factor. AB - Early pregnancy factor (EPF) is a secreted protein with growth regulatory and immunomodulatory properties. It functions as an autocrine growth factor for tumour cells and as an autocrine or paracrine growth factor for regenerating normal cells. Anti-EPF antibodies have demonstrable anti-tumour activity and, as a result, hybridomas which produce such antibodies are unstable. In this study, the phage display antibody techniques have been investigated as a means of producing recombinant anti-EPF antibodies. Mice were immunised with synthetic peptides which correspond to the N or C terminal regions of EPF, and their splenic tissue was used to make combinatorial antibody libraries. The Fab repertoire was displayed on the surface of phage and panned over recombinant EPF. Reactive Fabs were identified by ELISA and their binding was characterised by BIAcore analysis and functional studies. Three libraries with a size of greater than 5x10(7)cfu were constructed and a total of 26 unique Fabs with specific reactivity against EPF were identified. Three Fabs were purified and of these one demonstrated strong EPF neutralising activity, one had intermediate activity and the other was not neutralising. Phage display has provided the means of circumventing the problems of anti-EPF hybridoma development and has resulted in the production of antibodies with potential applications in the diagnosis of pregnancy and the diagnosis and therapy of cancer. PMID- 11033031 TI - Site-specific and directional gene replacement mediated by Cre recombinase. AB - A novel method for the site-specific introduction of genes into eukaryotic cells using the prokaryotic Cre-LoxP recombination system is presented. Cre recombinase catalyzes recombination between two LoxP sites or between two mutant LoxP 511 sites. However, recombination is not catalyzed between a LoxP and a LoxP 511 site. We now demonstrate that it is possible to catalyze accurate exchange between two DNA segments each flanked by a LoxP and a LoxP 511 site. In the example presented, expression of the Cre recombinase resulted in the replacement of a murine IgA constant region gene with a LoxP site at the 5' end and a LoxP 511 site at the 3' end by a human IgA constant region gene flanked by the same wild type and mutant LoxP sites. This method provides a novel approach for the site-specific substitution of specific genes. PMID- 11033033 TI - Isolation of endothelial cells from murine tissue. AB - The isolation and long-term culture of murine endothelial cells (ECs) has often proven a difficult task. In this paper we describe a quick, efficient protocol for the isolation of microvascular endothelial cells from murine tissues. Murine lung or heart are mechanically minced and enzymatically digested with collagenase and trypsin. The single cell suspension obtained is then incubated with an anti CD31 antibody, anti-CD105 antibody and with biotinylated isolectin B-4. Pure EC populations are finally obtained by magnetic bead separation using rat anti-mouse Ig- and streptavidin-conjugated microbeads. EC cultures are subsequently expanded and characterised. The surface molecule expression by the primary cultures of murine EC obtained from lung and heart tissue is analysed and compared to that of a murine endothelioma and of primary cultures of murine renal tubular epithelial cells. The phenotype and morphology of these cultures remain stable over 10-15 passages in culture, and no overgrowth of contaminating cells of non-endothelial origin is observed at any stage. PMID- 11033032 TI - Laboratory protocols for the identification of Th cell epitopes on self-antigens in mice with systemic autoimmune diseases. AB - T cells play a critical role in both the immunological and clinical manifestations of systemic autoimmune diseases such as systemic lupus erythematosus (SLE). Although in normal mice multiple T cell epitopes have been characterized in several self-proteins, there is little information on the fine specificity of autoreactive T cells in lupus model mice and humans. In SLE-prone mice and humans, the only Th cell epitopes identified at the molecular level in self-antigens concern histones and nucleosomes, and the 70-kD U1-snRNP protein. T cell characterization in certain autoimmune mice such as MRL lpr/lpr and NZB/NZW mice has been largely impaired by their hyporesponsiveness in response to mitogen and minimal IL-2 secretion. In addition, MRL lpr/lpr mice also develop lymphadenopathy characterized by the progressive accumulation of functionally immature CD4(-) CD8(-) T cells. It is therefore important to optimize the methods used to measure T cell proliferation and cytokine production ex vivo in order to identify minimal activation in the presence of appropriate antigen. The protocol described in this article has been used for identifying in young MRL lpr/lpr and NZB/NZW mice a CD4(+) T cell epitope in the murine 70-kD U1-RNP protein. PMID- 11033035 TI - Changes and additions to the editorial board PMID- 11033034 TI - Human immunoglobulin variable region gene analysis by single cell RT-PCR. AB - This protocol describes application of single cell reverse transcription polymerase chain reaction (RT-PCR) to the study of human immunoglobulin V region usage. The procedure begins with separation of peripheral blood mononuclear cells (PBMC) from human blood. The PBMC are stained with the B cell selective marker, anti-CD19. Stained B cells are sorted by flow cytometry and deposited, consecutively, one cell into each of an array of tubes. cDNA for one or more antibody variable regions (VH and/or VL) is synthesized with a primer (or primers) complementary to sequence(s) within the constant region (Cmu, Cgamma, Ckappa and/or Clambda). The cDNA is used as template for PCR amplification with gene or gene family specific primers. A second PCR is then performed with two nested primers to increase both the specificity and quantity of V region PCR products. The purified PCR products are sequenced directly and aligned to V region germline database and the Genbank database. Single cell RT-PCR is a fast and convenient way to analyze V region gene expression. It avoids the bias that may be introduced into V region cDNA library construction by the presence of highly variable levels of mRNA in different cells. The PCR products are obtained in quantities that can be cloned into bacterial expression vectors for production of recombinant V region protein domains. PMID- 11033036 TI - Penetrating keratoplasty and glaucoma. AB - Glaucoma remains the leading cause of blindness after penetrating keratoplasty. Post-keratoplasty glaucoma was originally described in 1969, and its management is still controversial. Recent developments in management include newer classes of drugs, surgical procedures, such as trabeculectomy with mitomycin-C, implantation of glaucoma drainage devices, and cyclodestructive procedures with Nd: YAG and diode lasers. However, the risk of graft failure continues to be high with all surgical procedures. PMID- 11033037 TI - Common errors in the use of magnetic resonance imaging for neuro-ophthalmic diagnosis. AB - The optimal utilization of magnetic resonance imaging in neuro-ophthalmic diagnosis is limited by errors in prescribing and interpreting scans. In a review of case material, we discovered four common prescriptive errors: 1) failure to apply a dedicated study, 2) inappropriate use of a dedicated study, 3) omission of intravenous contrast, and 4) omission of specialized sequences. The four common interpretive errors were the following: 1) failure to detect the lesion because of misleading clinical information, 2) rejection of a clinical diagnosis because an expected imaging abnormality was absent, 3) assumption that a striking imaging abnormality accounted for the clinical abnormality, and 4) failure to consider the lack of clinical specificity of imaging abnormalities. Many of these errors could be avoided by improved communication between clinicians and radiologists. PMID- 11033038 TI - The role of oxidative stress in the pathogenesis of age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of blind registration in the developed world, and yet its pathogenesis remains poorly understood. Oxidative stress, which refers to cellular damage caused by reactive oxygen intermediates (ROI), has been implicated in many disease processes, especially age-related disorders. ROIs include free radicals, hydrogen peroxide, and singlet oxygen, and they are often the byproducts of oxygen metabolism. The retina is particularly susceptible to oxidative stress because of its high consumption of oxygen, its high proportion of polyunsaturated fatty acids, and its exposure to visible light. In vitro studies have consistently shown that photochemical retinal injury is attributable to oxidative stress and that the antioxidant vitamins A, C, and E protect against this type of injury. Furthermore, there is strong evidence suggesting that lipofuscin is derived, at least in part, from oxidatively damaged photoreceptor outer segments and that it is itself a photoreactive substance. However, the relationships between dietary and serum levels of the antioxidant vitamins and age-related macular disease are less clear, although a protective effect of high plasma concentrations of alpha tocopherol has been convincingly demonstrated. Macular pigment is also believed to limit retinal oxidative damage by absorbing incoming blue light and/or quenching ROIs. Many putative risk-factors for AMD have been linked to a lack of macular pigment, including female gender, lens density, tobacco use, light iris color, and reduced visual sensitivity. Moreover, the Eye Disease Case-Control Study found that high plasma levels of lutein and zeaxanthin were associated with reduced risk of neovascular AMD. The concept that AMD can be attributed to cumulative oxidative stress is enticing, but remains unproven. With a view to reducing oxidative damage, the effect of nutritional antioxidant supplements on the onset and natural course of age-related macular disease is currently being evaluated. PMID- 11033039 TI - Pseudo-pseudotumor. AB - The initial diagnostic impression in a patient with a "hot orbit" may evolve as more laboratory and radiographic information becomes available. We present a patient in whom nearly the entire differential diagnosis of painful orbitopathy was considered before the correct diagnosis was made, after the patient had improved and all paraclinical data, including magnetic resonance imaging, were available. PMID- 11033040 TI - Theoretical limits to visual performance. AB - Wavefront sensors and scanning laser technology are enabling the correction of the aberrations of the eye. The effects of aberrations on visual performance are reviewed, and the theoretical limit of visual performance is predicted to understand the ultimate endpoint of these new technologies. A schematic eye model that incorporates diffraction, chromatic aberration, photopic response, the Stiles-Crawford effect, and pupil size is ray-traced to determine its limiting optical properties. These properties are compared to the detection requirements of the retina and brain to determine the theoretical limit of foveal vision. The theoretical limits on foveal vision are found to be between 20/12 and 20/5, depending on pupil diameter. It is concluded that emerging refractive surgery technologies may provide substantial increases in visual performance. PMID- 11033041 TI - A bruital headache and double vision. AB - A 44-year-old woman presented with a painful abducens nerve palsy in the left eye. Examination revealed a white, quiet eye and an orbital bruit without proptosis. Magnetic resonance imaging demonstrated abnormal ipsilateral dural based enhancement. Angiography confirmed a posterior draining carotid-cavernous fistula. Symptoms resolved spontaneously in approximately 8 months. The classification and treatment options for carotid-cavernous fistula are discussed. PMID- 11033042 TI - Really painful double vision. AB - A 67-year-old woman with a history of chronic headache and recent removal of two squamous cell lesions from her forehead presented with left facial pain and diplopia. A diagnosis of Tolosa-Hunt syndrome was made based on clinical presentation and imaging studies. When the patient did not respond to steroids, further studies were done, including biopsy, which revealed perineural spread of squamous cell carcinoma. PMID- 11033043 TI - Cataract development after trabeculectomy with mitomycin C: a 1-year study, by L. 'Daugeliene, T. Yamamoto, and Y. Kitazawa. Jpn J Ophthalmol 44:52-7, 2000. PMID- 11033044 TI - Neuronal correlates of amblyopia in the visual cortex of macaque monkeys with experimental strabismus and anisometropia, by L. Kiorpes, D. Kiper, L. O'Keefe, J. Cavanaugh, and A. Movshon. J neuroscience 18:6411-24, 1998. PMID- 11033045 TI - Environmental and heritable factors in the causation of cancer: analyses of cohorts of twins from Sweden, Denmark, and Finland, by P. Lichtenstein, N.V. Holm, P.K. Verkasalo, A. Iliadou, J. Kaprio, M. Koskenvuo, E. Pukkala, A. Skytthe, and K. Hemminki. N Engl J Med 343:78-84, 2000. PMID- 11033046 TI - "Numerous new regulatory peptides" PMID- 11033047 TI - Singular contributions of fish neuroendocrinology to mammalian regulatory peptide research. AB - During the past 20 years, several bioactive peptides have been identified in teleost fishes that subsequently have been shown to play important regulatory roles in mammalian physiology. The urophysis, corpuscles of Stannius and Brockmann body are anatomical structures particular to fish that have no obvious counterpart in mammals. Extracts and/or cDNA libraries prepared from these tissues have been used to identify for the first time urotensin II (U-II), urotensin-I (U-I), stanniocalcin and glucagon-like peptide-1 (GLP-1). Although U II and U-I were originally regarded as exclusively the products of the teleost urophysis, the peptides have a wide phylogenetic distribution across the vertebrate lineage, including mammals. U-II is localized to motor neurones in the human spinal cord and is a potent vasoconstrictor that may be implicated in the pathogenesis of heart failure. The human ortholog of urotensin-I is urocortin which is synthesized in selected regions of the brain and is the endogenous ligand for the CRF type 2 receptor. Urocortin is believed to important in mediating the effects of stress on appetite. Stanniocalcin is involved in maintaining calcium and phosphate homeostasis in teleost fish. An ortholog of stanniocalcin has a widespread distribution in mammalian tissues and is postulated to regulate renal phosphate excretion and to protect neurons against damage during cerebral ischemia. The biological actions and therapeutic potential of GLP-1 in humans are now fully appreciated but the peptide was first identified as a domain in a preproglucagon cDNA prepared from anglerfish Brockmann bodies. In contrast to mammalian preproglucagons, GLP-1 is present in anglerfish preproglucagon as the bioactive, truncated sequence [corresponding to human GLP 1(7-37)] rather than the inactive, N-terminally extended form [corresponding to GLP-1(1-37)]. Failure to appreciate the significance of this fact retarded progress in the field for several years. PMID- 11033048 TI - Helicobacter pylori infection and gastrin and cyclooxygenase expression in gastric and colorectal malignancies. AB - Helicobacter pylori, infecting more than 50% of the world population, results in gastritis, usually located in the antral portion of the stomach, accompanied by hypergastrinemia, the key factor in gastric and colorectal carcinogenesis. Excessive mucosal cell proliferation for many years may eventually result in gastric atrophy, cell mutation and transformation of gastric mucosal cells into gastrin-producing cells, which also express gastrin receptors serving to stimulate cell proliferation and tumor growth. These processes may be completed by the expression of cyclooxygenase-2 (COX-2) as an inflammation enzyme to release excessive amounts of PGE(2), leading to further proliferation, reduction in apoptosis, angiogenesis and tumor growth. H. pylori eradication results in complete regression of MALT lymphoma and subsequent normalisation of excessive gastrin release and COX-2 expression. Reduction of gastrin by active immunisation (gastrimmune), blocking of gastrin receptors with specific blockers and suppression of COX-2 might be helpful in inhibiting tumor growth and invasion. PMID- 11033049 TI - Pathways of inflammation and cell death in the lung: modulation by vasoactive intestinal peptide. AB - The pathogenesis of tissue injury in disease is a complex process that is only partially understood. We have investigated different models of acute lung injury, representing the clinical entity known as the acute respiratory distress syndrome, and tested their possible modulation by the neuropeptide vasoactive intestinal peptide (VIP). Three major mechanisms of injury appear to be involved in many of these models as common denominators: (1) activation of nuclear transcriptions factor NFkappaB; (2) apoptotic cell death; and (3) excitotoxic phenomena, due to activation of N-methyl-D-aspartate glutamate receptors. These pathogenetic mechanisms and pathways are logical targets of therapeutic intervention. Protection by VIP against lung injury, and against related forms of injury/cell death of neuronal cells and heart muscle, is attributable, in large measure, to the ability of VIP to suppress these mechanisms, and to additional anti-inflammatory and anti-oxidant actions. Finally, a hypothesis is presented for survival-promoting pathways that can be augmented by VIP and the related pituitary adenylyl cyclase-activating peptide. PMID- 11033050 TI - Gastric endocrine cells: types, function and growth. AB - The history of gastric endocrine cells identification and functional characterization is briefly outlined. An up to date classification of such cells is given. Present status of histopathological, histochemical, ultrastructural and molecular investigations on gastric endocrine hyperplasia and neoplasia is summarized and briefly discussed. PMID- 11033051 TI - Gastrin as a growth factor in the gastrointestinal tract. AB - The peptide hormone gastrin, released from antral G cells, is known to stimulate the synthesis and release of histamine from ECL cells in the oxyntic mucosa via CCK-2 receptors. The mobilized histamine induces acid secretion by binding to the H(2) receptors located on parietal cells. Recent studies suggest that gastrin, in both its fully amidated and less processed forms (progastrin and glycine-extended gastrin), is also a growth factor for the gastrointestinal tract. In this article, we review the recent evidence (including those from the transgenic and knockout mice) for the trophic targets of both the amidated and less processed forms of gastrin in the gastrointestinal tract, pancreas and liver. It has been established that the major trophic effect of amidated gastrin is for the oxyntic mucosa of stomach, where it causes increased proliferation of gastric stem cells and ECL cells, resulting in increased parietal and ECL cell mass. There is insufficient evidence to support that amidated gastrin is a trophic factor for the rest of gastrointestinal tract, exocrine pancreas and liver. On the other hand, the major trophic target of the less processed gastrin (e.g. glycine extended gastrin) appears to be the colonic mucosa. There is no evidence to suggest that it is trophic for the stomach. It remains to be examined whether the rest of gastrointestinal tract, pancreas and liver are the trophic targets by glycine-extended gastrin and progastrin. PMID- 11033052 TI - Gut hormones as pharmaceuticals. From enteroglucagon to GLP-1 and GLP-2. PMID- 11033053 TI - Principles of tachykininergic co-transmission in the peripheral and enteric nervous system. AB - The tachykinins substance P (SP) and neurokinin A (NKA) are synthesized and released from nerves in the peripheral and enteric nervous system (PNS and ENS). They act as nonadrenergic noncholinergic (NANC) excitatory transmitters in mammalian airways, and the genitourinary and gastrointestinal tract. At the postjunctional level, both NK(1) (SP-preferring) and NK(2) (NKA-preferring) receptors are often co-expressed by target cells innervated by TKergic nerves. Thus an issue of duplication seems to exists with regard to peripheral tachykininergic co-transmission, the duplication involving both messengers (the peptides) and effectors (the receptors). By using receptor selective antagonists it has been possible to dissect the relative contribution of different receptors to TKergic co-transmission: the available results indicate that multiple arrangements exist involving both summation, cooperation and specialization of different messengers/effectors in producing the overall response. PMID- 11033054 TI - The multiple actions of angiotensin II in atherosclerosis. AB - Angiotensin II (Ang II), the effector peptide of the renin-angiotensin system, has been implied in the pathogenesis of atherosclerosis on various levels. There is abundant experimental evidence that pharmacological antagonism of Ang II formation by angiotensin converting enzyme inhibition or blockade of the cellular effects of Ang II by angiotensin type 1 receptor blockade inhibits formation and progression of atherosclerotic lesions. Angiotensin promotes generation of oxidative stress in the vasculature, which appears to be a key mediator of Ang II induced endothelial dysfunction, endothelial cell apoptosis, and lipoprotein peroxidation. Ang II also induces cellular adhesion molecules, chemotactic and proinflammatory cytokines, all of which participate in the induction of an inflammatory response in the vessel wall. In addition, Ang II triggers responses in vascular smooth muscle cells that lead to proliferation, migration, and a phenotypic modulation resulting in production of growth factors and extracellular matrix. While all of these effects contribute to neointima formation and development of atherosclerotic lesions, Ang II may also be involved in acute complications of atherosclerosis by promoting plaque rupture and a hyperthrombotic state. Accordingly, Ang II appears to have a central role in the pathophysiology of atherosclerosis. PMID- 11033055 TI - Cholecystokinin and panic disorder--three unsettled questions. AB - The serendipitously discovered panicogenic effect of the cholecystokinin fragment, the C-terminal tetrapeptide amide (CCK-4), has suggested that the widespread network of CCK neurons and corresponding CCK-B receptors in the brain are in some way involved in pathogenesis panic disorders in man. Two decades of research have now established that exogenous CCK-4 in a reproducible, dose dependent and sensitive manner indeed evokes panic attacks in both healthy subjects and at even lower doses in anxiety patients. But several questions about the molecular mechanisms by which endogenous CCK peptides may precipitate panic attacks remain to be answered. This review focuses on three immediate questions. (1) Does endogenous CCK-4 exist? (2) Is the panicogenic effect mediated only through CCK-B receptors? (3) Are measurements of CCK peptides in cerebrospinal fluid of use in elucidating the pathogenesis and/or diagnosis? This review concludes that the answers to these questions may further the understanding of panic disorder substantially, and hence contribute to improved diagnosis and therapy of the disease. PMID- 11033056 TI - The neurobiology of urocortin. AB - Urocortin (UCN) is a recently isolated 40 amino acid-containing neuropeptide that is the second endogenous mammalian ligand for the corticotropin-releasing factor (CRF) receptors. While UCN and CRF both display a similar high affinity for the CRF(1) receptor, the affinity of UCN for the CRF(2) receptor is more than 10-fold higher than that of rat/human CRF. UCN mRNA expression is highest in the Edinger Westphal nucleus and lateral superior olive, with the most prominent terminal fields found in the lateral septum. Because of the higher relative affinity of UCN for the CRF(2) receptor and the corresponding neuroanatomical distribution of the highest density of UCN expression and innervation to brain regions preferentially expressing the CRF(2) receptor subtype, it has been hypothesized that UCN is the preferred endogenous ligand for the CRF(2) receptor. Following central administration, UCN has been demonstrated to produce behavioral and physiological effects that are qualitatively similar to CRF. Quantitatively, however, UCN appears to be a more potent suppressor of ingestive behavior (food and water intake) and a less potent inducer of anxiogenic behavior than CRF. PMID- 11033057 TI - Pathway specific expression of neuropeptides and autonomic control of the vasculature. AB - In this article, we review the immunohistochemical evidence for the pathway specific expression of co-existing neuropeptides in autonomic vasomotor neurons, and examine the functional significance of these expression patterns for the autonomic regulation of the vasculature. Most final motor neurons in autonomic vasomotor pathways contain neuropeptides in addition to non-peptide co transmitters such as catecholamines, acetylcholine and nitric oxide. Neuropeptides also occur in preganglionic vasomotor neurons. The precise combinations of neuropeptides expressed by neurons in vasomotor pathways vary with species, vascular bed, and the level within the vascular bed. This applies to both vasoconstrictor and vasodilator pathways. There is a similar degree of variation in the expression of neuropeptide receptors in the vasculature. Consequently, the contributions of different peptides to autonomic vasomotor control are closely matched to the functional requirements of specific vascular beds. This arrangement allows for a high degree of precision in vascular control in normal conditions and has the potential for considerable plasticity under pathophysiological conditions. PMID- 11033058 TI - Bayliss and Starling and the nascence of endocrinology. PMID- 11033059 TI - Neurotensin: peptide for the next millennium. AB - Neurotensin is an endogenous tridecapeptide neurotransmitter (pGlu-Leu-Tyr-Glu Asn-Lys-Pro-Arg-Arg-Pro-Try-Ile-Leu-OH) that was discovered by Carraway and Leeman in bovine hypothalami in the early 1970s. Since then this peptide has been the subject of a multitude of articles detailing discoveries related to its activity, receptors, localization, synthesis, and interactions with other systems. This review article does not intend to summarize again all the history of this fascinating peptide and its receptors, since this has been done quite well by others. The reader will be directed to these other reviews, where appropriate. Instead, this review attempts to provide a summary of current knowledge about neurotensin, why it is an important peptide to study, and where the field is heading. Special emphasis is placed on the behavioral studies, particularly with reference to agonists, antagonists, and antisense studies, as well as, the interaction of neurotensin with other neurotransmitters. PMID- 11033060 TI - Cytotoxicity due to corrosion of ear piercing studs. AB - It is well known that allergic and/or inflammatory reactions can be elicited from the use of gold-coated studs, particularly the type used for piercing ears, since they are left in contact with body fluids until the puncture heals. Inasmuch as gold is known as a non-toxic element, other elements of the substrate material may be responsible for some allergies. Therefore, characteristics of the coating, such as defects that expose the substrate to the human skin or body fluids, play an important role in the development of skin sensitization. In this study, the cytotoxicity of commercial studs used for ear piercing and laboratory-made studs was determined in a culture of mammalian cells. The corrosion performance of the studs was investigated by means of weight loss measurements and electrochemical impedance spectroscopy. The elements that leached out into the medium were also analysed by instrumental neutron activation analysis. Further, the surfaces of the studs were examined by scanning electron microscopy and analysed by energy dispersive spectroscopy to identify defects and reaction products on the surface, both before and after their exposure to the culture medium. The stud which showed lower corrosion performance resulted in higher cytotoxicity. Ti showed no cytotoxicity and high corrosion resistance, proving to be a potential material for the manufacture of ear piercing studs. PMID- 11033061 TI - Effects of model inducers on thyroxine UDP-glucuronosyl-transferase activity in vitro in rat and mouse hepatocyte cultures. AB - Thyroxine (T(4))-UDP-glucuronosyltransferase (UGT) activity was measured directly in cultured male Sprague-Dawley rat and OF-1 mouse hepatocyte monolayers. The activity of T(4)-UGT (pmol/min/g liver) in vitro in hepatocyte cultures was, after 24 hr in culture, equivalent to that previously measured in vivo in rat and mouse liver microsomes (Viollon-Abadie et al., 1999). A progressive decline in T(4)-UGT activity occurred over time in both rat and mouse hepatocyte cultures. Treatment of cultures with various model inducers such as phenobarbital (PB), beta-naphthoflavone (NF) and clofibric acid (CLO) induced a strong increase in T(4)-UGT activity in rat hepatocyte monolayers. In addition, and as expected from available in vivo data, treatment of rat hepatocyte cultures with NF also increased p-nitrophenol (PNP)-UGT activity and treatment with PB or CLO increased bilirubin (Bili)-UGT activity. In contrast, T(4)-UGT activity in mouse hepatocyte monolayers was not affected by the treatments, neither were PNP- and Bili- UGT activities. These in vitro data confirm our previous in vivo observations that these inducers increase rat but not mouse liver T(4)-UGT activities (Viollon Abadie et al., 1999). The present study thus demonstrates that hepatocyte monolayers are appropriated for the evaluation and inter-species comparison of the effects of xenobiotics on T(4)-UGT activities. PMID- 11033062 TI - Red cell haemolysis test as an in vitro approach for the assessment of toxicity of karanja oil. AB - The karanja tree grows in parts of India and Australia. The oil from seed kernels was found to be toxic to animals. The annual potential availability of the oil is around 135,000 tons in India. In order to use it for beneficial purposes, it is necessary to detoxify the oil. In the present study, the oil was assessed for toxicity by the red cell haemolysis test and estimating the LDH in the supernatant. The non-lipid constituents were isolated from raw oil by aqueous methanol extraction. The raw oil and the non-lipid fraction were found to haemolyse the red cells with release of LDH, whereas the extracted oil did not show such a manifestation. There was a good correlation between haemolytic activity and LDH released from cells. These findings were further confirmed with in vivo studies where the raw and extracted karanja oils showed 100% and nil mortality in rats dosed orally at 10 and 20 ml/kg body weight, respectively. This haemolysis test can be used as an in vitro method to predict toxicity and to monitor the detoxification of the oils prior to use in in vivo studies for toxicological evaluation. The fatty acid composition of the raw and extracted karanja oils showed no difference. PMID- 11033063 TI - In vitro human T cell sensitization to haptens by monocyte-derived dendritic cells. AB - We previously reported that in vitro primary sensitization of hapten-specific T cells by cultured human epidermal Langerhans cells (LC) provides an alternative approach to discriminate strong contact sensitizers from irritants (Krasteva et al., 1996; Moulon et al., 1993). However, this LC-based immunoassay was limited by the availability of human skin samples. In the present study, we used monocyte derived dendritic cells (DC) to analyse the autologous proliferative T cell response to several allergens. Monocytes were purified from the peripheral blood of healthy donors and cultured for 6-8 days in the presence of GM/CSF and IL-4 and then for 2 days in the presence of GM/CSF and TNFalpha. The resulting cells exhibited the phenotype of mature DC, as assessed by the strong expression of HLA DR, CD80, CD83 and CD86 antigens. We showed that trinitrophenyl (TNP)-treated mature DC induced a significant T cell proliferative response in all experiments, while fluorescein isothiocyanate (FITC) gave positive results in about half of them. The prohaptens eugenol and isoeugenol induced significant proliferation in one out of eight and in four out of 12 experiments, respectively. Interestingly, in 16 assays T cells never proliferated in the presence of sodium lauryl sulfate (SLS)-treated DC. Thus, this in vitro model allows discrimination between strong contact sensitizers and irritants. It might be very useful, therefore, for restriction of animal experimentation. PMID- 11033064 TI - Cryopreservation of precision-cut rat liver slices using a computer-controlled freezer. AB - Precision-cut liver slices are frequently used to study hepatic toxicity and metabolism of xenobiotics in vitro. Successful cryopreservation techniques will enhance an efficient and economic use of scarcely available (human) liver tissue. For primary hepatocytes, slow freezing has been accepted as the best approach towards successful cryopreservation. For slices, however, no agreement exists on the optimal way of cryopreservation and both slow and fast freezing techniques have been reported. The aim of the present study was to determine the applicability of a computer-controlled slow freezing technique for the cryopreservation of (rat) liver slices. Thus far, this technique has not been described in detail. Our studies confirmed that slow freezing was most successful in the cryopreservation of primary rat hepatocytes. Based on this observation, the slow freezing technique was applied to the cryopreservation of rat liver slices. Directly after thawing, slice viability was between 60 and 100% of fresh values, depending on the parameter determined. However, after additional culturing, slice viability was reduced. This decrease in slice viability was more pronounced in comparison to primary hepatocytes. In conclusion, the slow freezing technique was confirmed to be a successful approach for the cryopreservation of primary rat hepatocytes, and was found to be of limited use for the cryopreservation of rat liver slices. PMID- 11033065 TI - Temperature-dependent vitellogenin-mRNA expression in primary cultures of rainbow trout (Oncorhynchus mykiss) hepatocytes at 14 and 18 degrees C. AB - In order to study the influence of temperature on vitellogenin gene and estrogen receptor gene expression in primary hepatocytes from rainbow trout (Oncorhynchus mykiss), cells were exposed to 17beta-estradiol, bisphenol-A and nonylphenol for 48 and 96 hr. Induction of vitellogenin-mRNA expression was detected in a non radioactive dot blot/RNAse protection assay and by RT-PCR. In the dot blot/RNAse protection assay, the estrogenic potentials of bisphenol-A and nonylphenol were about 10(4)- to 10(5)-fold and 10(5)-fold lower than that of 17beta-estradiol, respectively. The relative estrogenic potential did not show any difference between 14 and 18 degrees C. In contrast, at 18 degrees C, RT-PCR analysis revealed increased amounts of vitellogenin- and estrogen receptor-mRNA after 12 and 24 hr of exposure to 17beta-estradiol, if compared to 14 degrees C. Owing to increased vitellogenin gene expression at 18 degrees C, the sensitivity of primary hepatocytes to 17beta-estradiol and bisphenol-A could be increased. PMID- 11033066 TI - The expression of surface markers on dendritic cells as indicators for the sensitizing potential of chemicals. AB - Novel approaches to testing of skin sensitizing chemicals have made use of immature dendritic cells (DCs) cultured from different hematopoietic progenitors. These cells resemble Langerhans cells (LCs), which are the most potent antigen presenting cells in the skin. Former research has focused on the phenotypic and functional changes of LCs after application of skin sensitizers. But it has proven difficult to isolate sufficient numbers of LCs from skin. This disadvantage is overcome by cultures of immature DCs providing high numbers of reactive cells. The aim of the present investigation was to test the response of DC cultures established from different blood donors to known sensitizers, an irritant and a vehicle. The sensitizers NiSO(4), dinitrochlorobenzene (DNCB), 2,4,6 trinitrobenzene sulfonic acid (TNBS), alpha-hexylcinnamaldehyde (Cinn) and eugenol (Eu) induced the up-regulation of the co-stimulatory molecule CD86, of intercellular adhesion molecule CD54 and of the HLA-DR antigen. The irritant sodium dodecyl sulfate (SDS) and the vehicle dimethyl sulfoxide (DMSO) had no effect. A high rate of responders within blood donors was found for NiSO(4), TNBS, Cinn and Eu, while DNCB was less effective. The augmentation of surface marker expression in dendritic cells obtained from peripheral human blood seems to be a promising readout in prescreening for strong and moderate sensitizers. This test could thus help to reduce animal numbers for in vivo testing. PMID- 11033067 TI - The quantitive chlorioallantoic membrane test using trypan blue stain to predict the eye irritancy of liquid scintillation cocktails. AB - The chorioallantoic membrane-trypan blue staining assay (CAM-TBS) is used to evaluate the potential ocular irritation caused by liquid scintillation cocktails constituted by complex mixtures, including surfactants and other potential irritants. The harmful effect of these substances is determined by the amount of trypan blue adsorbed by the CAM. In the membrane previously treated with the scintillation liquids this amount was concentration dependent only in certain cases, irrespective of the water solubility of the mixtures. In general, it showed a high correlation (r=0.986) with the scores obtained in the Draize eye irritation test. In the present study, only two cocktails presented values of trypan blue adsorption higher than those recorded for their eye irritation in vivo, irrespective of the solvent nature. Unlike the classical HET-CAM procedure, this assay is objective and allows the evaluation of opaque and coloured substances without interfering in the determination of irritancy. Despite these advantages, the method is not suitable for complex mixtures of products that induce ocular irritation in small quantities. PMID- 11033068 TI - Heavy metal cytotoxicity studied by electron probe X-ray microanalysis of cultured rat hepatocytes. AB - Cytotoxicity of the heavy metals gold, mercury, thallium and lead was studied by measuring the intracellular element distribution of cultured rat hepatocytes by energy dispersive electron probe X-ray microanalysis of freeze-dried cryosections in a scanning transmission electron microscope. Exposure of the cells to aqueous solutions containing heavy metal ions in concentrations reaching a critical concentration caused increase of intracellular sodium and chloride content accompanied or followed by decrease of intracellular potassium content. Thus, the intracellular potassium/sodium ratio drastically decreased from control values of approximately 10 to values below 1 before changes of cell morphology became visible. In experiments with gold or mercury the decrease of the potassium/sodium ratio was preceded by transient cytoplasmic increase of sulfur and phosphorus. Heavy metal concentrations exceeding the critical concentration also caused an increase of cytoplasmic calcium concentration and finally decay of the cell structure. Cytotoxicity of heavy metals was found to increase in the order Pb, Au, Tl, Hg. Cytotoxic effects by Au, Tl or Hg in moderate concentrations were reduced by simultaneous addition of Zn or Pb to the culture medium. The results obtained prove electron probe X-ray microanalysis of cryosections as a sensitive probe of cell viability. PMID- 11033069 TI - Induction of drug metabolism by nuclear receptor CAR: molecular mechanisms and implications for drug research. AB - Recent findings indicate that induction of drug metabolism is regulated by activation of specific members of the nuclear receptor gene family. This minireview deals with the mechanisms by which phenobarbital and phenobarbital type chemicals induce cytochrome P450 and other genes, and summarises the knowledge on the role of the constitutively active receptor CAR in the induction process. The potential implications of CAR-mediated induction for drug research and possible uses of CAR are also discussed. PMID- 11033070 TI - Multidrug resistance (MDR) in cancer. Mechanisms, reversal using modulators of MDR and the role of MDR modulators in influencing the pharmacokinetics of anticancer drugs. AB - In recent years, there has been an increased understanding of P-glycoprotein (P GP)-mediated pharmacokinetic interactions. In addition, its role in modifying the bioavailability of orally administered drugs via induction or inhibition has been also been demonstrated in various studies. This overview presents a background on some of the commonly documented mechanisms of multidrug resistance (MDR), reversal using modulators of MDR, followed by a discussion on the functional aspects of P-GP in the context of the pharmacokinetic interactions when multiple agents are coadministered. While adverse pharmacokinetic interactions have been documented with first and second generation MDR modulators, certain newer agents of the third generation class of compounds have been less susceptible in eliciting pharmacokinetic interactions. Although the review focuses on P-GP and the pharmacology of MDR reversal using MDR modulators, relevance of these drug transport proteins in the context of pharmacokinetic implications (drug absorption, distribution, clearance, and interactions) will also be discussed. PMID- 11033071 TI - Synthesis and pharmacological evaluation of novel heterotricyclic acylhydrazone derivatives, designed as PAF antagonists. AB - This paper describes the synthesis and the antiplatelet properties of new heterotricyclic N-acylhydrazone derivatives (7a-e), structurally analogous to known hetrazepinic PAF antagonists, exploring molecular hybridization as a tool for molecular designing. The synthetic route employed to access compounds (7a-e) used, as starting material, the previously described methyl 3-hydroxy-8-methyl-6 phenyl-6H-pyrazolo[3,4-b]thieno[2, 3-d]pyridine-2-carboxylate derivative. The results from inhibitory effects of these novel acylhydrazone derivatives (7a-e) upon PAF-induced platelet aggregation, indicated that all compounds present a significant antithrombotic profile. PMID- 11033072 TI - Effects of specific bile acids on c-fos messenger RNA levels in human colon carcinoma Caco-2 cells. AB - Bile acids may play a role in the pathogenesis of intestinal inflammation by activating the signalling pathways that control cell proliferation, among other cell systems. We investigated the action of different bile acids, particularly chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA), on steady-state and transcriptional regulation of the protooncogene c-fos, involved in the regulation of cell proliferation and differentiation, in colon carcinoma Caco-2 cells. Specific bile acids had a stimulatory effect of on the expression of c-fos mRNA. This proved to be concentration- and time-dependent and may be partly due to an increase in the rate of transcription of the corresponding gene rather than to any change in the stability of mRNA. In Caco-2 cells exposed to 250 microM CDCA for 1 h a maximal increase of c-fos mRNA ( approximately 2.5-fold induction over the control) was observed; deoxycholic acid (DCA; 250 microM) and lithocholic acid (LCA; 250 microM) were less effective (approximately 2-fold induction over the control). UDCA and cholic acid (CA) did not modify c-fos gene expression in this cell line. Finally, we investigated the role of protein kinase C (PKC) in transcriptional regulation of the c-fos gene by bile acids. Although induction of c-fos by 12-O-tetradecanoyl 13-acetate (10 nM), a potent PKC activator, was completely antagonised by bis-indolyl-maleimide I (1 microM); only about 40% of the bile acid-mediated rise in c-fos mRNA was blocked. Thus it appears that PKC, as well as other signalling pathways, is involved in CDCA-, DCA- and LCA-induced c-fos gene expression. PMID- 11033073 TI - Characterization of insulin-resistance: role of receptor alteration in insulin dependent diabetes mellitus, essential hypertension and cardiac hypertrophy. AB - Insulin-resistance is associated with a number of disease states such as diabetes, syndrome X, and hypertension. These situations may be coupled to insulin-resistance through the insulin signaling system as a common pathway. The purpose of this study was to investigate the receptor binding alterations in streptozotocin-induced diabetic rats, spontaneously hypertensive rats and aortocaval shunted rats (eccentric cardiac hypertrophy). A physical model describing a 1:1 stoichiometry of ligand binding with its receptor is proposed describing reversible binding of [(125)I]insulin or [(125)I]IGF-1 at the microvascular endothelial as well as with the cardiac myocytes after CHAPS treatment. Analysis of the collected effluents are curve-fitted with a conservation equation and a first-order Bessel function which allowed the calculation of the forward binding constants (k(n)), the reversible constants (k( n)), the dissociation constants (k(d)) and the residency time constants (tau). The results showed that streptozotocin-induced diabetic rats showed insulin resistance through alterations in the kinetics of insulin receptor binding. The normotensive controls of the spontaneously hypertension rats (SHR) carry themselves insulin-resistant receptors whose binding to insulin worsens in the hypertensive SHR. Negative cooperativity between insulin-like growth factor IGF-1 and insulin receptors could be a causative factor predisposing for insulin resistance in the aortocaval shunted rats to insulin resistance. The defects may be occurring at the receptor level in insulin-dependent diabetes mellitus, Wistar Kyoto rats and spontaneously hypertensive rats. In conclusion, alterations in the kinetics of insulin binding to its receptor seem to play a central role for the initiation of insulin-resistance during the various pathophysiological states. PMID- 11033074 TI - Impact of particle density and initial volume on mathematical compression models. AB - In the calculation of the coefficients of compression models for powders either the initial volume or the particle density is introduced as a normalising factor. The influence of these normalising factors is, however, widely different on coefficients derived from the Kawakita, Walker and Heckel equations. The problems are illustrated by investigations on compaction profiles of 17 materials with different molecular structures and particle densities. It is shown that the particle density of materials with covalent bonds in the Heckel model acts as a key parameter with a dominating influence on the derived apparent yield pressures. Substances with ionic bonds or macromolecular materials with amorphous structures do not show this dependency on the particle density. The influence of density is demonstrated by performing non-linear regression on the Heckel equation where the optimal particle density is estimated. The a parameter in the Kawakita equation is not influenced to a greater degree by variation in the initial volume. PMID- 11033075 TI - Region-dependent disappearance of vinblastine in rat small intestine and characterization of its P-glycoprotein-mediated efflux system. AB - This study was aimed to characterize the absorption behavior of vinblastine (VLB), a well-known substrate of P-glycoprotein (P-gp), from rat small intestine, especially focusing on the regional-dependence of its efflux mediated by P-gp. VLB disappeared from duodenal and ileal loops of male Wistar rats fairly rapidly (30-60% in 30 min). In contrast, its disappearance from the jejunal loop was almost negligible and in some rats >100% of the jejunal dose was recovered. The radioactivity derived from [3H]VLB, which was absorbed from duodenum and ileum, was detected in the jejunal region. The jejunal appearance of radioactivity was increased when unlabeled VLB was present in the region in advance. The basolateral-to-apical transport of [3H]VLB across Caco-2 cell monolayers was greater when unlabeled VLB was added to the apical medium than when VLB-free buffer was applied to the apical side. When verapamil or cyclosporin A, potent modulators of P-gp, was added to the apical medium together with unlabeled VLB, enhanced basolateral-to-apical transport of [3H]VLB was disappeared. It is suggested that VLB absorption is strongly restricted by P-gp, especially in the jejunal region of the rat small intestine, and that the secretory transport via intestinal P-gp may be subject to trans-stimulation. Moreover, intravenously administered methylprednisolone and intramuscularly administered progesterone significantly enhanced the absorption of VLB, suggesting that parenterally administered P-gp modulators could influence the intestinal absorption of P-gp substrates. PMID- 11033076 TI - 4-toluenesulfonylureido derivatives of amines, amino acids and dipeptides: a novel class of potential antitumor agents. AB - The screening of a series of arylsulfonylureido derivatives of amines (such as histamine, or dopamine), aliphatic/aromatic amino acids (such as Gly, beta-Ala, Val, Lys, Arg, Phe, Tyr, DOPA, etc.) and dipeptides (such as GlyGly, beta-AlaHis) led to the identification of three derivatives that possess tumor growth inhibitory properties against several leukemia, non-small cell lung, ovarian, melanoma, colon, CNS, renal, and breast cancer cell lines in vitro. The new derivatives were prepared by reaction of 4-toluenesulfonyl isocyanate with (protected) amines, amino acids or dipeptides. The mechanism of antitumor action with these new derivatives is not known at the moment but it may imply uncoupling of mitochondria, as for the structurally related diarylsulfonylurea sulofenur, an investigational anticancer agent. PMID- 11033077 TI - Bioadhesive potential of gliadin nanoparticulate systems. AB - The objective of this work was to prepare, characterise and evaluate the adhesive potential of gliadin nanoparticulate carriers. Firstly, lectin-nanoparticle conjugates were obtained by the carbodiimide (CDI) covalent binding of Dolichos biflorus lectin (DBA) to the surface of gliadin nanoparticles (NP) containing carbazole (as a model lipophilic drug). The DBA binding efficiency was favoured in mild acidic conditions. Similarly, a CDI concentration of about 0.63 mg/mg nanoparticles, acting during at least 1 h, provided binding efficiencies of about 50% bulk lectin. Under optimised experimental conditions, the DBA conjugates showed a size of around 500 nm and the amount of loaded carbazole and the DBA content were calculated to be around 15 and 23.5 microg/mg, respectively. The bioadhesive activity of NP and DBA conjugates was determined in samples of small and large rat intestinal mucosa. The amount of adsorbed NP was calculated to be around 8 and 4 g/m(2) in the small and large intestine, respectively. This high capacity to interact with the mucosa may be explained by gliadin composition. In fact, gliadin is rich in neutral and lipophilic residues. Neutral amino acids can promote hydrogen bonding interactions with the mucosa, while the lipophilic components can interact with the biological tissue by hydrophobic interactions. The bioadhesive activity of DBA conjugates was calculated to be about 2 g/m(2) in the small intestine and greater than 4 g/m(2) in the caecum and distal colon. These degrees of interaction were always significantly higher than those obtained with controls. Finally, DBA did not provide the specificity for interaction with Peyer's patches. In summary, gliadin nanoparticles show a high capacity of non specific interaction with the intestine, whereas DBA binding to the surface of these carriers provided a greater specificity for colonic mucosa. PMID- 11033078 TI - Transdermal delivery of levosimendan. AB - The aim of this study was to determine if transdermal penetration of levosimendan, a novel positive inotropic drug, could be enhanced and controlled by formulation modifications. Penetration of levosimendan across human epidermis in vitro was determined using abdominal excised skin and diffusion cells. Predicted steady-state plasma concentrations of levosimendan were estimated using permeabilities and pharmacokinetic parameters of levosimendan. For penetration enhancement we used different pH values, co-solvents, cyclodextrins, surfactants, penetration enhancers, liposomes, and iontophoresis. Sodium lauryl sulfate, ethanol, oleic acid, and soya phosphatidylcholine or their combinations clearly increased levosimendan permeation across the skin in vitro. Iontophoresis was also an efficient method to increase transdermal permeation of levosimendan. A hydrophilic co-solvent/penetration enhancer is needed to achieve better permeability of levosimendan across the skin. In conclusion, transdermal delivery of levosimendan can be significantly increased by formulation modification. Based on kinetic calculations, therapeutic plasma concentrations may be achievable transdermally. PMID- 11033079 TI - Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC7806: an integrated peptide-polyketide synthetase system. AB - BACKGROUND: Blooms of toxic cyanobacteria (blue-green algae) have become increasingly common in the surface waters of the world. Of the known toxins produced by cyanobacteria, the microcystins are the most significant threat to human and animal health. These cyclic peptides are potent inhibitors of eukaryotic protein phosphatases type 1 and 2A. Synthesized nonribosomally, the microcystins contain a number of unusual amino acid residues including the beta amino polyketide moiety Adda (3-amino-9-methoxy-2,6, 8-trimethyl-10-phenyl-4,6 decadienoic acid). We have characterized the microcystin biosynthetic gene cluster from Microcystis aeruginosa PCC7806. RESULTS: A cluster spanning 55 kb, composed of 10 bidirectionally transcribed open reading frames arranged in two putative operons (mcyA-C and mcyD-J), has been correlated with microcystin formation by gene disruption and mutant analysis. Of the 48 sequential catalytic reactions involved in microcystin synthesis, 45 have been assigned to catalytic domains within six large multienzyme synthases/synthetases (McyA-E, G), which incorporate the precursors phenylacetate, malonyl-CoA, S-adenosyl-L-methionine, glutamate, serine, alanine, leucine, D-methyl-isoaspartate, and arginine. The additional four monofunctional proteins are putatively involved in O-methylation (McyJ), epimerization (McyF), dehydration (McyI), and localization (McyH). The unusual polyketide amino acid Adda is formed by transamination of a polyketide precursor as enzyme-bound intermediate, and not released during the process. CONCLUSIONS: This report is the first complete description of the biosynthesis pathway of a complex cyanobacterial metabolite. The enzymatic organization of the microcystin assembly represents an integrated polyketide-peptide biosynthetic pathway with a number of unusual structural and enzymatic features. These include the integrated synthesis of a beta-amino-pentaketide precursor and the formation of beta- and gamma-carboxyl-peptide bonds, respectively. Other features of this complex system also observed in diverse related biosynthetic clusters are integrated C- and N-methyltransferases, an integrated aminotransferase, and an associated O-methyltransferase and a racemase acting on acidic amino acids. PMID- 11033080 TI - Aminoacyl-SNACs as small-molecule substrates for the condensation domains of nonribosomal peptide synthetases. AB - BACKGROUND: Nonribosomal peptide synthetases (NRPSs) are large multidomain proteins that catalyze the formation of a wide range of biologically active natural products. These megasynthetases contain condensation (C) domains that catalyze peptide bond formation and chain elongation. The natural substrates for C domains are biosynthetic intermediates that are covalently tethered to thiolation (T) domains within the synthetase by thioester linkages. Characterizing C domain substrate specificity is important for the engineered biosynthesis of new compounds. RESULTS: We synthesized a series of aminoacyl-N acetylcysteamine thioesters (aminoacyl-SNACs) and show that they are small molecule substrates for NRPS C domains. Comparison of rates of peptide bond formation catalyzed by the C domain from enterobactin synthetase with various aminoacyl-SNACs as downstream (acceptor) substrates revealed high selectivity for the natural substrate analog L-Ser-SNAC. Comparing L- and D-Phe-SNACs as upstream (donor) substrates for the first C domain from tyrocidine synthetase revealed clear D- versus L-selectivity. CONCLUSIONS: Aminoacyl-SNACs are substrates for NRPS C domains and are useful for characterizing the substrate specificity of C domain-catalyzed peptide bond formation. PMID- 11033081 TI - Potent active site-directed inhibition of steroid sulphatase by tricyclic coumarin-based sulphamates. AB - BACKGROUND: There is now abundant evidence that inhibition of steroid sulphatase alone or in conjunction with inhibition of aromatase may enhance the response of postmenopausal patients with hormone-dependent breast cancer to this type of endocrine therapy. Additionally, sulphatase inhibition has been proposed to be of potential therapeutic benefit in the immune system and for neuro-degenerative diseases. After the finding that our first highly potent active site-directed steroid sulphatase inhibitor, oestrone-3-O-sulphamate (EMATE), was highly oestrogenic, we proposed non-steroidal coumarin sulphamates such as 4 methylcoumarin-7-O-sulphamate (COUMATE) as alternative non-steroidal steroid sulphatase inhibitors. In this work, we describe how tricyclic coumarin-based sulphamates have been developed which are even more potent than COUMATE, are non oestrogenic and orally active. We also discuss potential mechanisms of action. RESULTS: 4-Ethyl- (4), 4-(n-propyl)- (6), 3-ethyl-4-methyl- (8), 4-methyl-3-(n propyl)coumarin-7-O-sulphamate (11); the tricyclic derivatives 665COUMATE (13), 666COUMATE (15), 667COUMATE (17), 668COUMATE (20) and the tricyclic oxepin sulphamate (22) were synthesised. In a placental microsome preparation, all of these analogues were found to be more active than COUMATE in the inhibition of oestrone sulphatase, with the most potent inhibitor being 667COUMATE which has an IC(50) of 8 nM, some 3-fold lower than that for EMATE (25 nM). In addition, 667COUMATE was also found to inhibit DHEA-sulphatase some 25-fold more potently than EMATE in a placental microsome preparation. Like EMATE, 667COUMATE acts in a time- and concentration-dependent manner, suggesting that it is an active site directed inhibitor. However, in contrast to EMATE, 667COUMATE has the important advantage of not being oestrogenic. In addition, we propose several diverse mechanisms of action for this active site-directed steroid sulphatase inhibitor in the light of recent publications on the crystal structures of human arylsulphatases A and B and the catalytic site topology for the hydrolysis of a sulphate ester. CONCLUSIONS: A highly potent non-steroidal, non-oestrogenic and irreversible steroid sulphatase inhibitor has been developed. Several mechanisms of action for an active site-directed steroid sulphatase inhibitor are proposed. With 667COUMATE now in pre-clinical development for clinical trial, this should allow the biological and/or clinical significance of steroid sulphatase inhibitors in the treatment of postmenopausal women with hormone-dependent breast cancer and other therapeutic indications to be fully evaluated. PMID- 11033082 TI - Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. AB - BACKGROUND: Glycogen synthase kinase-3 (GSK-3) is a serine/threonine protein kinase, the activity of which is inhibited by a variety of extracellular stimuli including insulin, growth factors, cell specification factors and cell adhesion. Consequently, inhibition of GSK-3 activity has been proposed to play a role in the regulation of numerous signalling pathways that elicit pleiotropic cellular responses. This report describes the identification and characterisation of potent and selective small molecule inhibitors of GSK-3. RESULTS: SB-216763 and SB-415286 are structurally distinct maleimides that inhibit GSK-3alpha in vitro, with K(i)s of 9 nM and 31 nM respectively, in an ATP competitive manner. These compounds inhibited GSK-3beta with similar potency. However, neither compound significantly inhibited any member of a panel of 24 other protein kinases. Furthermore, treatment of cells with either compound stimulated responses characteristic of extracellular stimuli that are known to inhibit GSK-3 activity. Thus, SB-216763 and SB-415286 stimulated glycogen synthesis in human liver cells and induced expression of a beta-catenin-LEF/TCF regulated reporter gene in HEK293 cells. In both cases, compound treatment was demonstrated to inhibit cellular GSK-3 activity as assessed by activation of glycogen synthase, which is a direct target of this kinase. CONCLUSIONS: SB-216763 and SB-415286 are novel, potent and selective cell permeable inhibitors of GSK-3. Therefore, these compounds represent valuable pharmacological tools with which the role of GSK-3 in cellular signalling can be further elucidated. Furthermore, development of similar compounds may be of use therapeutically in disease states associated with elevated GSK-3 activity such as non-insulin dependent diabetes mellitus and neurodegenerative disease. PMID- 11033083 TI - FR900482 class of anti-tumor drugs cross-links oncoprotein HMG I/Y to DNA in vivo. AB - BACKGROUND: Overexpression of the high-mobility group, HMG I/Y, family of chromatin oncoproteins has been implicated as a clinical diagnostic marker for both neoplastic cellular transformation and increased metastatic potential of several human cancers. These minor groove DNA-binding oncoproteins are thus an attractive target for anti-tumor chemotherapy. FR900482 represents a new class of anti-tumor agents that bind to the minor groove of DNA and exhibit greatly reduced host toxicity compared to the structurally related mitomycin C class of anti-tumor drugs. We report covalent cross-linking of DNA to HMG I/Y by FR900482 in vivo which represents the first example of a covalent DNA-drug-protein cross link with a minor groove-binding oncoprotein and a potential novel mechanism through which these compounds exert their anti-tumor activity. RESULTS: Using a modified chromatin immunoprecipitation procedure, fragments of DNA that have been covalently cross-linked by FR900482 to HMG I/Y proteins in vivo were polymerase chain reaction-amplified, isolated and characterized. The nuclear samples from control cells were devoid of DNA fragments whereas the nuclear samples from cells treated with FR900482 contained DNA fragments which were cross-linked by the drug to the minor groove-binding HMG I/Y proteins in vivo. Additional control experiments established that the drug also cross-linked other non-oncogenic minor groove-binding proteins (HMG-1 and HMG-2) but did not cross-link major groove binding proteins (Elf-1 and NFkappaB) in vivo. Our results are the first demonstration that FR900482 cross-links a number of minor groove-binding proteins in vivo and suggests that the cross-linking of the HMG I/Y oncoproteins may participate in the mode of efficacy as a chemotherapeutic agent. CONCLUSIONS: We have illustrated that the FR class of anti-tumor antibiotics, represented in this study by FR900482, is able to produce covalent cross-links between the HMG I/Y oncoproteins and DNA in vivo. The ability of this class of compounds to cross link the HMG I/Y proteins in the minor groove of DNA represents the first demonstration of drug-induced cross-linking of a specific cancer-related protein to DNA in living cells. We have also demonstrated that FR900482 cross-links other minor groove-binding proteins (HMG-1 and HMG-2 in the present study) in vivo; however, since HMG I/Y is the only minor groove-binding oncoprotein presently known, it is possible that these non-histone chromatin proteins are among the important in vivo targets of this family of drugs. These compounds have already been assessed as representing a compelling clinical replacement for mitomycin C due to their greatly reduced host toxicity and superior DNA interstrand cross linking efficacy. The capacity of FR900482 to cross-link the HMG I/Y oncoprotein with nuclear DNA in vivo potentially represents a significant elucidation of the anti-tumor efficacy of this family of anticancer agents. PMID- 11033085 TI - Peptidyl transferase: ancient and exiguous. AB - The finding that the universal ribosomal peptidyl transferase is an RNA enzyme casts new light on its ancient origins, on the use of transition state analogues for ribozymes, and on the role of selection-amplification in studies of molecular evolution. PMID- 11033084 TI - Lipoic acid-derived amphiphiles for redox-controlled DNA delivery. AB - BACKGROUND: Intracellular release of free DNA from the vector complex is one of the critical steps limiting the efficiency of non-viral gene delivery. The complex should be stable enough to prevent DNA degradation but it should be destabilized inside the cell to allow DNA release and transcription. Destabilization and degradation of synthetic vectors is also required to reduce their cytotoxicity and augment the life-time of transfected cells. RESULTS: Here we describe new cationic amphiphiles made from the natural pro-vitamin, lipoic acid, that reversibly binds and releases DNA, depending on the redox state of the lipoate moieties. In the oxidized state these amphiphiles condense DNA into homogeneous spherical particles, which, upon reduction, swell into DNA toroids with subsequent release of free DNA. Complex reduction and DNA release can be induced by various thiols as well as enzymatically, by thioredoxin reductase. Transfection with amphiphile-DNA complexes in vitro shows a several fold increase of transgene expression compared with DOTAP, and can be further augmented by attachment of the nucleus-targeting peptide to the amphiphile. The increase of transfection efficiency results from GSH- and NAD(P)H-dependent complex reduction and release of free DNA inside the cells. CONCLUSIONS: The present work demonstrates the principle of a redox-controlled gene delivery system that uses the reversibility of thiol-disulfide exchange reaction. Our data suggest that the efficiency of synthetic vectors can be augmented by their controlled destabilization inside the cells. Being formed from the natural non-toxic compound lipoic acid, these cationic amphiphiles provide a new promising class of synthetic vectors for gene delivery. PMID- 11033086 TI - Life after worms: Lynx Therapeutics, Inc. PMID- 11033087 TI - Differential antagonism of endomorphin-1 and endomorphin-2 spinal antinociception by naloxonazine and 3-methoxynaltrexone. AB - To determine the role of spinal mu-opioid receptor subtypes in antinociception induced by intrathecal (i.t.) injection of endomorphin-1 and -2, we assessed the effects of beta-funaltrexamine (a selective mu-opioid receptor antagonist) naloxonazine (a selective antagonist at the mu(1)-opioid receptor) and a novel receptor antagonist (3-methoxynaltrexone) using the paw-withdrawal test. Antinociception of i.t. endomorphins and [D-Ala(2), MePhe(4), Gly(ol)(5)]enkephalin (DAMGO) was completely reversed by pretreatment with beta funaltrexamine (40 mg/kg s.c.). Pretreatment with a variety of doses of i.t. or s.c. naloxonazine 24 h before testing antagonized the antinociception of endomorphin-1, -2 and DAMGO. Judging from the ID(50) values of naloxonazine, the antinociceptive effect of endomorphin-2 was more sensitive to naloxonazine than that of endomorphin-1 or DAMGO. The selective morphine-6beta-glucuronide antagonist, 3-methoxynaltrexone, which blocked endomorphin-2-induced antinociception at each dose (0.25 mg/kg s.c. or 2.5 ng i.t.) that was inactive against DAMGO, did not affect endomorphin-1-induced antinociception but shifted the dose-response curve of endomorphin-2 3-fold to the right. These findings may be interpreted as indicative of the existence of a novel mu-opioid receptor subtype in spinal sites, where antinociception of morphine-6beta-glucuronide and endomorphin-2 are antagonized by 3-methoxynaltrexone. The present results suggest that endomorphin-1 and endomorphin-2 may produce antinociception through different subtypes of mu-opioid receptor. PMID- 11033088 TI - Temporal changes in neuronal dropout following inductions of lithium/pilocarpine seizures in the rat. AB - Estimates of neuronal dropout for approximately 100 structures as defined by Paxinos-Watson were completed for brains of male Wistar albino rats between 1 and 50 days after status epilepticus was evoked by a single systemic injection of lithium and pilocarpine. Sample estimates of neuronal loss were strongly correlated with direct measures of cell density. The most extensive immediate damage occurred within the substantia nigra reticulata, CA1 field of the hippocampus, the piriform cortex and the reuniens and paratenial nuclei of the thalamus. Neuronal dropout continued in many other structures over a 50-day period. Structures that showed the greatest 2-deoxyglucose (2-DG) uptake during discrete seizures and waxing and waning seizures within the early stages of status epilepticus but the least 2-DG uptake at the time of late continuous spiking and fast spiking with pauses [Neuroscience 64 (1995) 1057, 1075] exhibited the most neuronal dropout. Relationships between the delay of injection of acepromazine (which facilitated survival) and the amount of damage suggested that the source of the process that results in permanent brain damage may originate within the region of the piriform cortices and its subcortices. PMID- 11033089 TI - The domain of brain beta-spectrin responsible for synaptic vesicle association is essential for synaptic transmission. AB - We have examined the interaction between synapsin I, the major phosphoprotein on the membrane of small synaptic vesicles, and brain spectrin. Using recombinant peptides we have localized the synapsin I attachment site upon the beta-spectrin isoform betaSpIISigmaI to a region of 25 amino acids, residues 211 through 235. This segment is adjacent to the actin binding domain and is within the region of the betaSpIISigmaI that we previously predicted as a candidate synapsin I binding domain based upon sequence homology. We used differential centrifugation techniques to quantitatively assess the interaction of spectrin with synaptic vesicles. Using this assay, high affinity saturable binding of recombinant betaSpIISigmaI proteins was observed with synaptic vesicles. Binding was only observed when the 25 amino acid synapsin I binding site was included on the recombinant peptides. Further, we demonstrate that antibodies directed against 15 amino acids of the synapsin I binding domain specifically blocked synaptic transmission in cultured hippocampal neurons. Thus, the synapsin I attachment site on betaSpIISigmaI spectrin comprises a approximately 25 amino acid segment of the molecule and interaction of these two proteins is an essential step for the process of neurotransmission. PMID- 11033090 TI - Cholinergic modulation of neocortical long-term potentiation in the awake, freely moving rat. AB - The neocortex has proven resistant to LTP induction using standard in vitro and acute, in vivo preparations. Because the neocortex is widely thought to be involved in long-term information storage, this resistance raises questions about the validity of LTP as a memory model. Recently, we have shown that the neocortex of freely moving rats reliably supports LTP, provided that the stimulation is spaced and repeated over days. The following experiments were designed to evaluate the neuromodulatory role played by cholinergic systems in the induction of LTP in this preparation. Chronically implanted rats received either low- or high-intensity LTP-inducing tetani in combination with the administration of either a cholinergic agonist or antagonist injected systemically. Potentiation was evidenced as amplitude changes in both early and late components of the evoked field potential, the former including population spikes. The cholinergic agonist facilitated LTP induction in the late component of both high- and low intensity groups. The cholinergic antagonist blocked LTP induction in the early component of the high-intensity group. The possibility that there are component specific modulatory effects of cholinergic agents on the induction of neocortical LTP is discussed. PMID- 11033091 TI - Improved bioavailability to the brain of glycosylated Met-enkephalin analogs. AB - The blood-brain barrier prevents the entry of many potentially therapeutic peptide drugs to the brain. Glycosylation has shown potential as a methodology for improving delivery to the CNS. Previous studies have shown improved bioavailability and improved centrally mediated analgesia of glycosylated opioids. In this study we investigate the effect of glycosylation on the cyclic opioid peptide [D-Cys(2,5),Ser(6),Gly(7)] enkephalin. The peptide was glycosylated on the Ser(6) via an O-linkage with various sugar moieties and alignments. The peptides were then investigated for receptor binding, physiochemical attributes, in situ brain uptake in female Sprague-Dawley rats and antinociception in male ICR mice. Glycosylation resulted in a slight decrease in affinity to the delta-opioid receptor, and mixed effect on binding to the mu opioid receptor. There was a significant decrease in lipophilicity resulting from glycosylation and a slight reduction in binding to bovine serum albumin. In situ perfusion showed that brain uptake was improved by up to 98% for several of the glycosylated peptides, and the nociceptive profiles of the peptides, in general, followed the rank order of peptide entry to the brain with up to a 39-fold increase in A.U.C. PMID- 11033092 TI - Organization of efferent neurons in area 19: the projection to extrastriate area 21a. AB - The organization of efferent neurons in area 19 of the cat was examined by bulk injections of retrograde tracers, WGA-HRP and CTX-Au, into extrastriate area 21a. In one case, the cortex was cut coronally and retrogradely labeled cells in area 19 were present in columnar register throughout layers 2 to 6, with the majority of labeled cells in layers 2/3. The number of columns per tissue section ranged from 0 to 4 and had a centre-to-centre spacing ranging from 0.6 to 0.9 mm. A few lightly labeled cells were found between the columns. In six other cases, the visual cortex was flattened, and cut in the tangential plane to reveal a pattern of irregular, widely spaced bands that were elongated in the mediolateral direction with a mean centre-to-centre spacing of 2.6 mm. The density of labeled cells within these bands fluctuated such that dense aggregates of cells were found, on average, at 0.9 mm intervals along the bands. This tangential heterogeneity in density, along with the patchy columnar staining witnessed in the coronal plane, suggests that the widely spaced efferent projection bands may have a patchy substructure with a spacing of approximately 1 mm. The pattern of efferent projection bands and its substructure in area 19 is reminiscent of the stripe-like organization of V2 found in primates. PMID- 11033093 TI - Differential transport of rat and human interleukin-1alpha across the blood-brain barrier and blood-testis barrier in rats. AB - Human interleukin-1alpha is transported across the murine blood-brain barrier (BBB) and blood-testis barrier (BTB) by a saturable transport system. Differences in the biological activity and binding of human IL-1 in mouse and rat brain raise the possibility of species differences in the transport of IL-1 across the BBB and BTB. We measured the transport of recombinant human 125I-IL-1alpha (I-huIL 1alpha) and rat 125I-IL-1alpha (I-ratIL-1alpha) across the rat BBB and BTB after intravenous injection using a sensitive in vivo technique and film autoradiography. I-ratIL-1alpha was found to cross the rat BBB and rat BTB at rates comparable to those reported previously for murine IL-1alpha in mice. Passage across the BBB was inhibited by the addition of unlabeled rat IL-1alpha, demonstrating saturable transport. In contrast, I-huIL-1alpha entered the brain of the rat much more slowly, and its entry was not inhibited by the addition of unlabeled human IL-1alpha. These results show that the rat interleukin-1 transporter, unlike the murine transporter, does not transport human IL-1alpha. This difference highlights the importance of species specificity in IL-1alpha transport and may partly explain the different physiological responses to exogenous human IL-1alpha among rodent species. PMID- 11033094 TI - Corticofugal inhibition compresses all types of rate-intensity functions of inferior collicular neurons in the big brown bat. AB - Recent studies have shown that the auditory corticofugal system modulates and improves signal processing in the frequency, time and spatial domains. In this study, we examine corticofugal modulation of rate-intensity functions of inferior collicular (IC) neurons of the big brown bat, Eptesicus fuscus, by electrical stimulation in the primary auditory cortex (AC). Cortical electrical stimulation compressed all types of rate-intensity functions so as to increase the slope but decrease the dynamic range of IC neurons. Cortical electrical stimulation also shifts the responsive intensity of IC neurons to higher levels. These data indicate that corticofugal modulation also improves subcortical signal processing in intensity domain. The implication of these findings to bat echolocation is discussed. PMID- 11033095 TI - Effects of nigrostriatal dopamine denervation on ionotropic glutamate receptors in rat caudate-putamen. AB - Changes in ionotropic glutamate NMDA, AMPA and KA receptor binding in rat caudate putamen were examined by quantitative in vitro receptor autoradiography 5 weeks after lesioning nigrostriatal dopaminergic projections. In this animal model of Parkinson's disease, density of binding in caudate-putamen increased at KA, but not NMDA or AMPA receptors. The findings indicate that nigrostriatal dopamine denervation can selectively enhance KA receptor levels in rat basal ganglia, suggest that KA receptors contribute to the pathophysiology of Parkinson's disease, and may suggest innovative treatments. PMID- 11033096 TI - Differential schwann cell migration in adult and old mice: an in vitro study. AB - The influence of aging on Schwann cell (SC) proliferation, migration and viability was studied in vitro. SCs were cultured in Ham F-10 medium enriched with 20% fetal calf serum (FCS), 40% FCS or collagen I gel plus 20% FCS. The migration of adult mice derived SCs was stimulated with FCS and collagen. With aging, SC migration, multiplication and viability decreased, indicating that ideal culturing conditions should be adjusted. PMID- 11033097 TI - Synaptic depression and neuronal loss in transiently acidic hippocampal slice cultures. AB - Acidosis is a rapid and inevitable event accompanying cerebral ischemia or trauma. We used hippocampal slice cultures to examine an immediate effect of acidosis, synaptic depression; and a delayed effect, neuronal loss. Exposure to low bicarbonate artificial cerebral spinal fluid (aCSF), pH 6.70 for 30 min at 32 degrees C, acidified intracellular pH from 7.31+/-0.12 to 6.53+/-0.08. Accompanying intracellular acidosis was a depression of synaptic responses. Both effects rapidly reversed after treatment with normal aCSF pH 7.35. Death analysis after acidosis treatment revealed no delayed neuronal loss. Increasing the duration of the acidosis to 60 min, however, induced irreversible synaptic depression and delayed neuronal loss. Increasing acidosis temperature to 37 degrees C acidified intracellular pH and depressed synaptic responses. Delayed neuronal loss was also observed. Acidosis using lactate aCSF, pH 6. 70 for 30 min at 32 degrees C acidified intracellular pH from 7. 19+/-0.13 to 6.43+/-0.07 and depressed synaptic responses. After reperfusion with lactate containing aCSF pH 7.35, intracellular pH recovered yet synaptic responses remained depressed and delayed neuronal loss was observed. This suggested that, for a 30-min treatment at 32 degrees C, lactate acidosis was neurotoxic while low bicarbonate acidosis was not. Increasing the duration or temperature of low bicarbonate acidosis induced neuronal loss. These data provide additional evidence that acidosis contributes to the neurotoxicity during stroke and trauma. PMID- 11033098 TI - Hypothermia as an adjunctive treatment for severe bacterial meningitis. AB - Brain injury due to bacterial meningitis results in a high mortality rate and significant neurologic sequelae in survivors. The objective of this study was to determine if the application of moderate hypothermia shortly after the administration of antibiotics would attenuate the inflammatory response and increase in intracranial pressure that occurs in meningitis. For this study we used a rabbit model of severe Group B streptococcal meningitis. The first component of this study evaluated the effects of hypothermia on blood-brain barrier function and markers of inflammation in meningitic animals. The second part of the study evaluated the effects of hypothermia on intracranial pressure, cerebral perfusion pressure and brain edema. This study demonstrates that the use of hypothermia preserves CSF/serum glucose ratio, decreases CSF protein and nitric oxide and attenuates myeloperoxidase activity in brain tissue. In the second part of this study we show a decrease in intracranial pressure, an improvement in cerebral perfusion pressure and a decrease in cerebral edema in hypothermic meningitic animals. We conclude that in the treatment of severe bacterial meningitis, the application of moderate hypothermia initiated shortly after antibiotic therapy improves short-term physiologic measures associated with brain injury. PMID- 11033099 TI - Infusion of 3alpha,5alpha-THP to the pontine reticular formation attenuates PTZ induced seizures. AB - Whether progesterone (P(4)) and its metabolite, 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP) have anti-seizure effects through actions in the pontine reticular formation (PRF) was investigated. Concentrations of P(4) and 3alpha, 5alpha-THP in the PRF were greater in proestrous and hormone-primed rats, that are typically more resistant to seizure-induction, than diestrous and males rats. Ovx, Long-Evans rats with unilateral microinjections into the PRF of 3alpha,5alpha-THP (5 microg/0.2 microl), but not P(4) (11 microg/0.2 microl) or vehicle (beta-cyclodextrin), had a greater latency and lower incidence of tonic clonic seizures induced by pentylenetetrazol (PTZ; 70 mg/kg, IP) administration. Infusions that missed the PRF were not effective. These data suggest 3alpha,5alpha-THP has anti-seizure effects in part through actions in the PRF. PMID- 11033100 TI - Reviewer's recommendations predict impact: why you have to send us your very best work. PMID- 11033101 TI - Cardiac work and efficiency. PMID- 11033102 TI - Cardiac gap junctions: good or bad? PMID- 11033103 TI - Decrease of delayed rectifier currents in the subacute phase of infarction. PMID- 11033104 TI - Calcium signalling in endothelial cells. AB - Vascular endothelial cells are ubiquitous for their presence in each and every vessel and unique for their multifunctional nature. A large number of endothelial functions depend to various extents on changes in intracellular Ca(2+) concentration. Reviewed are endothelial Ca(2+) stores, Ca(2+) channels, and in out-in Ca(2+) signalling events, from ligand-binding on the plasma membrane into depletion of intracellular Ca(2+) stores and therefrom out to transplasmalemmal Ca(2+) entry that is of prime importance for many endothelial functions. Special emphasis is placed on mechanisms regulating store-operated Ca(2+) entry including a Ca(2+) influx factor, the vesicle secretion-like model, the conformational coupling model, the membrane potential, cytochrome P450, protein tyrosine kinase, myosin light chain kinase and nitric oxide. PMID- 11033105 TI - Pathophysiology and treatment of haemodynamic instability in acute pulmonary embolism: the pivotal role of pulmonary vasoconstriction. AB - Acute massive pulmonary embolism has a high mortality rate. Fatal haemodynamic deterioration is caused by an acute increase in pulmonary vascular resistance. Traditionally, the degree of mechanical obstruction of the pulmonary vasculature by the embolic thrombus is considered to be the major determinant of this increase in right ventricular afterload. However, there is evidence to suggest that another factor plays an important role, since there is a marked discrepancy between the haemodynamic manifestations of acute pulmonary embolism and the degree of mechanical obstruction. Historic studies indicate that this discrepancy is largely explained by pulmonary vasoconstriction caused by vasoactive mediators, released mainly by activated platelets. Thromboxane-A(2) and serotonin are probably the two most important pulmonary vasoconstrictors in this context. Antagonising their effects dramatically increases tolerance to experimental pulmonary embolism in animals. In humans, this concept should eventually find its way into clinical practice. In the future, acute massive pulmonary embolism could be treated with antagonists to pulmonary vasoconstrictors, or with direct pulmonary vasodilators. PMID- 11033106 TI - Delayed rectifier K currents have reduced amplitudes and altered kinetics in myocytes from infarcted canine ventricle. AB - OBJECTIVE: The rapid (I(Kr)) and slow (I(Ks)) components of delayed rectifier currents play an important role in determining the cardiac action potential configuration. Abnormalities in their function may contribute to arrhythmogenesis under pathological conditions. We studied the effects of myocardial infarction on I(Kr) and I(Ks) in canine ventricular myocytes and their molecular basis. METHODS: Infarct zone myocytes (IZs) were isolated from a thin layer of surviving epicardium overlying an infarct 5 days after a total occlusion of the left anterior descending (LAD) coronary artery. Normal myocytes (NZs) were isolated from the corresponding region of control hearts for comparison. Currents were recorded under the whole-cell patch clamp conditions. RESULTS: Both I(Kr) and I(Ks) current densities were reduced in IZs versus NZs. Kinetic analysis further suggests an acceleration of I(Kr) activation and I(Ks) deactivation. RNase protection assays were used to quantify the mRNA levels of I(Kr) and I(Ks) channel subunits (dERG, dIsK and dKvLQT1) in tissue immediately adjacent to the region where myocytes were isolated. mRNA levels of all three subunits were reduced 2 days after LAD occlusion (by 48+/-9%, 68+/-5%, and 45+/-4% for dERG, dIsK and dKvLQT1, respectively, n=8 each). By day 5, the dKvLQT1 message returned to control while those of dERG and dIsK remained reduced (by 52+/-7% and 76+/-6%, respectively). CONCLUSIONS: The decrease in I(Kr) and I(Ks) amplitudes and changes in their kinetics in infarcted tissue might be due to a decrease in functional channels and/or changes in their subunit composition. Heterogeneous changes in I(Kr) and I(Ks) in infarcted hearts may impact on the effects of varying heart rate or neurohumoral modulation on repolarization. PMID- 11033107 TI - Comparative effects of azimilide and ambasilide on the human ether-a-go-go related gene (HERG) potassium channel. AB - OBJECTIVE: To evaluate the effects of azimilide and ambasilide on the biophysical properties of the human-ether-a-go-go-related (HERG) channel. METHODS: HERG was stably transfected into Chinese hamster ovary (CHO-K1) cells and currents were measured using a whole cell, voltage-clamp technique. RESULTS: Azimilide had a 'dual effect', inhibiting current at voltage steps above -40 mV and augmenting current at -40 and -50 mV. Tail current inhibition following a step to +30 mV did not vary with temperature (IC(50) 610 nM at 22 degrees C and 560 nM at 37 degrees C). The agonist effect at -50 mV was concentration-dependent and correlated with a hyperpolarizing shift in the V(1/2) of activation (r=0.98, P<0.05). Time constants of inactivation were faster and there was a -10 mV shift in the V(1/2) of steady state inactivation suggestive of open and inactivated state binding. By comparison, ambasilide inhibited HERG channels with lower potency (IC(50) 3.6 microM), in a voltage- and time-dependent but frequency-independent manner (0.03 1 Hz). Ambasilide had no effect on activation or inactivation gating but prolonged both fast and slow components of deactivation consistent with unbinding from the open state. The net effect of both drugs was similar during a voltage ramp which simulated a cardiac action potential. CONCLUSIONS: Inhibition of HERG channels by azimilide and ambasilide exhibits a similar time and voltage dependence. While both exhibit affinity for the open state, azimilide also binds to inactivated channels. PMID- 11033108 TI - The role of L-type calcium current in the generation of repolarization-induced contraction in cardiac myocytes. AB - OBJECTIVE: Early experiments into the arrhythmogenic transient inward current frequently showed apparent coupling of this current to repolarization from a depolarizing voltage clamp step. Calcium transients have subsequently been shown to couple to such repolarization and are the result of calcium release from the sarcoplasmic reticulum. We have investigated whether this phenomenon is due to calcium entry via non-inactivated calcium channels or to voltage-activated SR release. METHODS: Voltage clamp steps were imposed on isolated guinea pig and rabbit cardiac myocytes. Calcium release was monitored by tracking cell contraction. L-type calcium current at the moment of repolarization was manipulated by the rapid application of 2 mM cadmium or 10 mM calcium. RESULTS: Repolarization-induced contraction was abolished by the rapid application of 2 mM cadmium immediately prior to repolarization, and was augmented by the rapid change of extracellular calcium concentration from 2 mM to 10 mM immediately prior to repolarization. There is no evidence of coupling of drive train-induced aftercontractions to repolarization from the final action potential of the drive train and 2 mM cadmium does not alter the appearance or timing of these aftercontractions. Simulation of phase 1 repolarization in the mammalian cardiac action potential decreases rather than increases twitch amplitude. CONCLUSION: Repolarization-induced contraction results from calcium entry through non inactivated calcium channels, not from voltage-activated release. It plays no physiological role in contributing to the stimulated twitch and no pathological role in generating drive train-induced aftercontractions. PMID- 11033109 TI - Cell-to-cell interaction prevents cell death in cultured neonatal rat ventricular myocytes. AB - OBJECTIVES: Loss of cardiac cells and the anatomical or functional remodeling of intercellular coupling occur under several pathological conditions. We have assessed the significance of intercellular coupling for cell death. METHODS AND RESULTS: Ventricular cells obtained from 1 day old Wistar rats were cultured. Apoptosis was detected by nick-end labeling. Cells were plated at low and high cell density (3x10(4)/ml and 12x10(4)/ml, respectively). Cultured myocytes died spontaneously by apoptosis in a time dependent manner. The increase of the apoptotic cell population in a culture with high cell density on day 4 (1+/-1.2%, n=4) was significantly lower than that in a culture with low cell density (20+/ 5.5%, n=4). The progression of apoptosis in the culture of low cell density was prevented in part after application of the medium extract from the culture of high cell density; the apoptotic cell population on day 6 decreased from 57+/ 8.0% (n=4) to 36+/-3.8% (n=4). Treatment of the cultured myocytes at high cell density with antisense oligonucleotide for connexin43 (Cx43) for 24 h on day 2 resulted in a significant decrease in Cx43 expression as judged by Western blot, dye transfer and immunocytochemistry using mouse monoclonal antibody for Cx43. In association with the down-regulation of Cx43, the progress of apoptosis was accelerated; the apoptotic cell population on day 5 in the antisense-treated cultures (27+/-5.7%, n=4) was significantly higher than the sense-treated cultures (5+/-1.1%, n=4). The effect of Cx43 antisense treatment to promote apoptosis was not reversed by application of high cell-density culture medium. CONCLUSIONS: These findings suggest that cell-cell communication through gap junction formation and some humoral factors play important roles in the survival of cultured myocytes. PMID- 11033110 TI - Myocyte contractile function is intact in the post-infarct remodeled rat heart despite molecular alterations. AB - OBJECTIVE: To investigate the cellular mechanisms underlying global and regional LV dysfunction in the post-infarct (MI) remodeled rat hearts. METHODS: LV remodeling and function were quantified by echocardiography, morphometry, in vivo hemodynamics, and isolated perfused heart studies in 6 weeks post-MI and sham operated rats. LV myocytes from sham and MI hearts were used for morphometric and functional studies. Myocyte contractile function and intracellular calcium kinetics were measured at different stimulation frequencies (0.2-2 Hz), temperatures (30 and 37 degrees C), and external viscous load (1, 15, 200 and 300 centipoise). Myocyte apoptosis was measured by DNA laddering; BCL-2, BAX, Na(+) Ca(2+) exchanger, and SERCA-2 proteins by western blot; and brain natriuretic peptide (BNP), SERCA-2 mRNA by RT-PCR. RESULTS: MI hearts were remodeled (Echo LV diameter 7.3+/-0.38 vs. 5.9+/-0.16 mm, P<0.03), and showed global (Echo % fractional shortening 30+/-2.4 vs. 58+/-3, P<0.001), and regional contractile dysfunction of non-infarcted myocardium (Echo % systolic posterior wall thickening 36+/-2 vs. 57+/-1.7, P<0.001). In vivo hemodynamic and isolated heart function studies confirmed depressed LV systolic and diastolic function and increased volumes. Whereas, myocytes isolated from infarcted hearts were remodeled (40% longer and 10% wider), their contractile function and calcium kinetics under basal conditions and at high stimulation frequency, temperature and viscous load were similar to sham myocytes. The mRNA for BNP was increased whereas that for SERCA-2 decreased, but the SERCA-2 protein was normal. Despite myocyte hypertrophy, ventricular septal thickness was reduced in infarcted hearts (2.2+/-0.1 vs. 2. 6+/-0.07 mm, P<0.01), and showed increased apoptosis. CONCLUSIONS: Myocytes from remote non-infarcted myocardium of the remodeled hearts have normal contractile function, despite structural remodeling and altered gene expression. Non-myocyte factors may be more important in genesis of contractile dysfunction in the remodeled heart, for up to 6 weeks after MI. PMID- 11033111 TI - Myofibroblasts in reperfused myocardial infarcts express the embryonic form of smooth muscle myosin heavy chain (SMemb). AB - OBJECTIVE: The purpose of this study is to examine the cellular content of healing myocardial infarcts and study the phenotypic characteristics of fibroblasts during scar formation utilizing a canine model of coronary occlusion and reperfusion. METHODS: Ischemia/Reperfusion experiments were performed in dogs undergoing 1 h of coronary occlusion followed by reperfusion intervals ranging from 5 h to 28 days. Fibrotic and control areas were studied using immunohistochemistry. RESULTS: The healing ischemic and reperfused myocardium demonstrated significant proliferative activity peaking after 3 to 7 days of reperfusion, predominantly in myofibroblasts. The numbers of proliferating cells decreased during the maturation phase of the scar (PCNA index: 13.7+/-2.25% at 5 days vs. 4.8+/-1.1% at 28 days; P<0.05, n=5). During the proliferative phase of healing (3-7 days) alpha-smooth muscle actin (alpha-SMAc) expression was markedly increased in the fibrotic areas. alpha-SMAc predominantly localized in myofibroblasts which were vimentin positive, smooth muscle myosin, calponin and desmin negative. We examined expression of smooth muscle myosin heavy chain isoforms in myofibroblasts infiltrating the healing areas and found a marked induction of the embryonal isoform of myosin heavy chain (SMemb) in alpha-SMAc positive spindle shaped cells in the border of the scar. Myofibroblasts did not express SM2, a marker for mature smooth muscle cells. In contrast myocardial arterioles were positive for SM2, but did not express SMemb. CONCLUSIONS: Healing myocardial infarcts undergo rapid changes in their content of myofibroblasts. During the proliferative phase fibroblasts undergo phenotypic changes leading to expression of contractile proteins such as alpha-SMAc, and production of SMemb, a marker for dedifferentiated smooth muscle cells. Expression of embryonic isoforms indicates dedifferentiation and allows the myofibroblast pool to serve as a versatile cell population, assuming different phenotypes depending on the physiological needs. PMID- 11033112 TI - Cellular FLIP is expressed in cardiomyocytes and down-regulated in TUNEL-positive grafted cardiac tissues. AB - OBJECTIVE: c-FLIP is a natural homologue of caspase 8, and may antagonize activation of death pathways mediated by FADD. c-FLIP is highly expressed in the heart, and a recent report suggests that c-FLIP may protect against certain types of myocyte death. The present study was designed to define the expression patterns of c-FLIP in the heart. METHODS: The expression pattern of c-FLIP in end stage human hearts, and rat cardiomyocyte grafting models was analyzed by in situ hybridization, immunohistochemistry and TUNEL assay. In addition, to determine whether Fas-dependent pathway is active in cardiomyocytes in vitro, we examined whether activated monocytes can kill neonatal cardiomyocytes in a co-culture system. RESULTS: c-FLIP mRNA and protein were abundantly expressed in normal cardiomyocytes from failing human heart. In animal models, c-FLIP protein was absent in TUNEL-positive grafted cardiomyocytes. Double staining demonstrated that c-FLIP-positive cells rarely had fragmented DNA, while TUNEL-positive cells rarely contained c-FLIP. Finally, activated monocytes induced death of neonatal rat cardiomyocytes via the Fas/FasL system. CONCLUSIONS: Loss of c-FLIP expression correlates with cardiomyocyte cell death. We hypothesize that diminished c-FLIP expression may predispose cardiomyocytes to apoptotic death. PMID- 11033113 TI - Effects of aging on the work output and efficiency of rat papillary muscle. AB - OBJECTIVES: This study aimed to investigate the effect of aging on the work output and efficiency of rat papillary muscle. METHODS: The mechanical and energetic properties of left ventricular papillary muscle preparations isolated from 6-, 15-, and 27- to 32-month-old Sprague-Dawley rats were measured in myothermic experiments at 27 degrees C at a stimulus frequency of 0.167 Hz. RESULTS: We found that the basal metabolism measured in quiescent papillary muscles was significantly reduced in the 27- to 32-month-old group (4.9 mWg(-1) compared to 7.7 and 7.0 mWg(-1) in the 6- and 15-month groups). In isotonic experiments, the work output (at a range of afterloads) was significantly depressed for the 27- to 32-month group being only 52% of the work output of the 6-month group. This outcome was due to a decrease in both the extent of muscle shortening only, 66% of 6- and 15-month data, and in the maximum force developed. The reduced work was accompanied by a parallel decrease in energy consumption (enthalpy) and hence, the net mechanical efficiency (work/active enthalpyx100%) was not altered. A force-length- area (FLA) analysis was applied to the isotonic data and an energy: FLA regression line was obtained for each preparation. We found that there were no significant differences in either the intercept or slope of the energy: FLA relation with age. Contractile efficiency (39+/-3%) in the 27- to 32-month group was not significantly different to that found in the 6-month (43+/-4%) or 15-month (40+/-3% group). CONCLUSION: There are no changes in the mechanical performance or efficiency of cardiac muscle from young (6-month-old) or adult (15-month-old) rats but in the aged and senescent rats (27-32-month-old) there is a pronounced decline in stress development and shortening ability leading to a fall in work output. Mechanical and contractile efficiency however remain unchanged in old age and the data resembles that obtained in pressure overload hypertrophy. PMID- 11033114 TI - Regional electrophysiological effects of left ventricular hypertrophy in isolated rabbit hearts under normal and ischaemic conditions. AB - OBJECTIVES: Left ventricular hypertrophy (LVH) has been reported to produce differential electrophysiological effects in isolated epicardial and endocardial cells. This study aimed to examine regional electrophysiological effects of LVH in normal and ischaemic conditions in the whole heart. METHODS: LVH was secondary to perinephritis-induced hypertension. Monophasic action potential duration (MAPD(90)), effective refractory period (ERP) and conduction delay were measured in paced, isolated working rabbit hearts either at one right ventricular and two left ventricular sites (apical and basal epicardium) or at three left ventricular sites (apical and basal epicardium, apical endocardium). The hearts were subjected to 30 min of regional ischaemia and 15 min of reperfusion. RESULTS: In non-ischaemic conditions, LVH produced uniform prolongation of MAPD(90) and ERP in the left ventricular epicardium, but not in the endocardium. After coronary artery occlusion, LVH significantly increased ischaemia-induced transepicardial dispersion of repolarisation, but not refractoriness. LVH did not affect arrhythmogenesis in either non-ischaemic or ischaemic conditions. CONCLUSIONS: Differential effects of LVH on epicardial and endocardial electrophysiological parameters are also observed in the whole heart. In addition, the sensitivity of hypertrophied myocardium to ischaemia is increased and leads to an increase in ischaemia-induced dispersion of repolarisation. However, neither dispersion of refractoriness nor arrhythmogenesis are affected by LVH in non-ischaemic or ischaemic conditions in this experimental model. PMID- 11033115 TI - Effects of lipids on the functional and metabolic recovery from global myocardial stunning in isolated rabbit hearts. AB - OBJECTIVES: High concentrations of free fatty acids may increase myocardial ischaemic damage. However, the administration of lipid emulsions during reperfusion improves the functional recovery of stunned myocardium. From this apparent controversy we hypothesise that the effect of lipids is related to the time of its administration: we compared the effects of pre- and post-ischaemic administration of Intralipid((R)) on stunned myocardium. We also examined the role of fatty acids and phospholipids, respectively, in the effect of lipid emulsions on stunned myocardium. METHODS: Myocardial stunning was produced by 15 min of ischaemia and 90 min of reperfusion in isolated blood perfused rabbit hearts. Intralipid((R)) was administered either prior to ischaemia or during reperfusion. Left ventricular pressure (LVP) and its first derivative (LVdP/dt) were measured to assess functional recovery. High energy phosphates were measured with HPLC. The effects of linoleic acid, phosphatidylcholine and their combination were also studied. RESULTS: Only when Intralipid((R)) was administered during reperfusion, it improved recovery from contractile function and increased high energy phosphate content in globally stunned myocardium. Both linoleic acid and phosphatidylcholine significantly improved myocardial function in stunned myocardium. CONCLUSIONS: The effect of lipids on the contractile performance and metabolic state of stunned myocardium depends mainly on the timing of its administration with regard to the ischaemia/reperfusion event. Both free fatty acids and phospholipids contribute to the beneficial effect of lipid emulsions on functional recovery of stunned myocardium. PMID- 11033116 TI - DNA-RNA chimeric hammerhead ribozyme to transforming growth factor-beta(1) mRNA inhibits the exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats. AB - OBJECTIVE: The purpose of this study was to develop DNA-RNA chimeric hammerhead ribozyme against transforming growth factor-beta(1) (TGF-beta(1)) mRNA as a gene therapy agent for arterial proliferative diseases. METHODS: A 38-base hammerhead ribozyme against rat TGF-beta(1) mRNA, to produce cleavage at the GUC sequence at nucleotide 825 according to the secondary structure of rat TGF-beta(1) mRNA was designed. To enhance its stability, we synthesized a DNA-RNA chimeric ribozyme with two phosphorothioate linkages at the 3'-terminal. We also synthesized a mismatch ribozyme with single base change in the catalytic loop region as a control. These ribozymes were delivered into rat vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats by lipofectin-mediated transfection, and their biological effects were investigated. RESULTS: According to in vitro cleavage studies, the synthetic ribozyme can cleave the synthetic substrate RNA into two RNA fragments. Chimeric ribozyme significantly inhibited DNA synthesis in VSMC from SHR but not in cells from WKY rats. Mismatch ribozyme showed only a little effect on growth of VSMC from SHR. Chimeric ribozyme significantly inhibited proliferation of VSMC from SHR; in contrast, the proliferation of VSMC from WKY rats was significantly increased by this chimeric ribozyme. Mismatch ribozyme did not affect proliferation of VSMC from either rat strain. Chimeric hammerhead ribozyme to rat TGF-beta(1) dose-dependently inhibited TGF-beta(1) mRNA expression detected by reverse transcription and polymerase chain reaction analysis in VSMC from both rat strains. Chimeric hammerhead ribozyme to rat TGF-beta(1) also dose dependently inhibited TGF-beta(1) protein production detected by Western blot analysis. CONCLUSIONS: The present results demonstrated that our designed DNA-RNA chimeric hammerhead ribozyme to TGF-beta(1) mRNA might be a useful gene therapy agent for hypertensive vascular diseases. PMID- 11033117 TI - Growth factor-induced phosphoinositide 3-OH kinase/Akt phosphorylation in smooth muscle cells: induction of cell proliferation and inhibition of cell death. AB - OBJECTIVE: The signaling pathways mediating proliferation and apoptosis in vascular smooth muscle cells (VSMC) are not well established. It has previously been shown that activation of the phosphoinositide 3-OH kinase (PI3K)/Akt pathway or the ERK 1/2 pathway can mediate anti-apoptotic function in different cell types. This study determined the specific contribution of the PI3K/Akt and ERK pathway in the regulation of apoptosis and proliferation of VSMC. METHODS AND RESULTS: Incubation of rat VSMC with FCS, insulin or IGF-1 time-dependently stimulated the phosphorylation of Akt, however FCS but not insulin or IGF-1 activated the MAP-kinase ERK 1/2. Moreover, insulin inhibited H(2)O(2)-induced apoptosis via the Akt pathway as demonstrated by pharmacological inhibition of the PI3K or overexpression of a dominant negative Akt mutant. In contrast, FCS inhibited H(2)O(2)-induced apoptosis via the Akt and also the ERK pathway. FCS, but not insulin or IGF-1 induced VSMC proliferation, suggesting that Akt activation is necessary but not sufficient for VSMC proliferation. FCS-induced proliferation of VSMC was only mediated via the Akt pathway and not the ERK pathway. CONCLUSIONS: These results define a link between cell proliferation and programmed cell death in VSMC via the same signal transduction pathway, namely activation of the serine/threonine kinase Akt, which may have significant implication for the development of vascular diseases or remodeling. PMID- 11033118 TI - Blockade of endothelin receptors markedly reduces atherosclerosis in LDL receptor deficient mice: role of endothelin in macrophage foam cell formation. AB - OBJECTIVE: We evaluated the direct effects of long-term blockade of ET(A) and ET(B) receptors using a mixed endothelin (ET) receptor antagonist, LU224332, in the low density lipoprotein receptor (LDL-R) knockout mouse model of atherosclerosis. METHODS: Four groups of LDL-R deficient mice were studied: control mice fed normal chow (group I); mice fed a high cholesterol (HC, 1.25%) diet alone (group II), HC fed animals treated with LU224332 (group III); and mice fed normal chow treated with the LU compound (group IV). All treatments were continued for 8 weeks at which time the animals were sacrificed and the aortae were removed and stained with oil red O. Atherosclerotic area (AA) was determined by quantitative morphometry and normalized relative to total aortic area (TA). RESULTS: Cholesterol feeding resulted in a marked increased in total plasma cholesterol ( approximately 15 fold) and widespread aortic atherosclerosis (AA/TA: group I: 0.013+/-0.007; group II: 0.33+/-0. 11; P<0.001). Atherosclerotic lesions were characterized by immunohistochemistry as consisting mainly of macrophages which also showed high levels of ET-1 expression. Treatment with ET antagonist significantly reduced the development of atherosclerosis (AA/TA: group III: 0.19+/-0.07, P<0.01 vs. group II), without altering plasma cholesterol levels and blood pressure. The direct effect of LU224332 on macrophage activation and foam-cell formation was determined in vitro using a human macrophage cell line, THP-1. Treatment of the THP-1 cells with LU224332 significantly reduced cholesterol ester and triacylglycerol accumulation and foam-cell formation on exposure to oxidized LDL (P<0.01 and P<0.05, respectively). CONCLUSION: We conclude that a nonselective ET receptor antagonist substantially inhibited the development of atherosclerosis in a genetic model of hyperlipidemia, possibly by inhibiting macrophage foam-cell formation, suggesting a role for these agents in the treatment and prevention of atherosclerotic vascular disease. PMID- 11033119 TI - Temporal regulation of endothelial ET-1 and eNOS expression in intact human conduit vessels exposed to different intraluminal pressure levels at physiological shear stress. AB - OBJECTIVE: By using a computerized vascular perfusion model, we investigated temporal effects of sub-acute pressure elevation on vasomotor behavior and expression of endothelin-1 (ET-1) and endothelial nitric oxide synthase (eNOS) in intact human conduit vessels. METHODS: Paired umbilical veins were perfused during 1.5, 3 and 6 h under high/low intraluminal pressure (40/20 mmHg) and at identical shear stress level of 10 dyn/cm(2). ET-1 and eNOS gene and protein expression was quantified with real-time reverse-transcribed polymerase chain reaction and quantitative immunohistochemistry, respectively. RESULTS: Pressure induced differential temporal regulation patterns of ET-1 and eNOS gene expression. During the high pressure condition, eNOS mRNA was upregulated after 3 h and leveled off after 6 h of perfusion, while ET-1 mRNA was elevated after 6 h perfusion. Immunohistochemistry verified synchronal changes at the protein level. Significant vasodilation was observed after 3 h in the high-pressure system. CONCLUSION: Thus, subacute pressure elevation exerts differential effects on the endothelial eNOS/ET-1 expression, which dynamically regulate the vasomotor tone. PMID- 11033120 TI - Effects of alertness on three-dimensional eye movements. AB - PURPOSE: To investigate effects of alertness on three-dimensional (3D) eye movements. METHODS: During drowsy and alert periods, 3D eye movements were recorded with dual search coils in three normal rhesus monkeys, and in two of these monkeys after placement of bilateral kainic acid lesions of the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF). RESULTS: When the three monkeys were in the normal state, the average plane thickness (standard deviation of all rotation vectors from the regressed plane) during drowsy periods (range, 1.2-1.9 degrees ) increased significantly compared with the plane thickness during alert periods (range, 0.7-0.9 degrees ) in the light (P <.05). The plane thickness increased significantly in the monkey with asymmetric bilateral riMLF-lesions (P <.05); however, the increase was not significant in the monkey with more symmetric bilateral riMLF-lesions. After the bilateral riMLF lesions, the plane thickness also increased with drowsiness in both monkeys (P <.02), as it had in the normal state. CONCLUSION: The implementation of Listing's law is independent of the presence of vertical-torsional burst neurons in the riMLF. The increase in thickness of Listing's plane during drowsy periods is not due to an imprecise signal from saccadic burst neurons to the 3D velocity-to position integrator. It is possible that this integrator itself controls Listing's law, depending on the state of alertness. PMID- 11033121 TI - Genome analysis with restriction endonucleases recognizing 4- or 5-base pair sequences of adenovirus type. AB - PURPOSE: To detect the genetic changes in adenovirus type 4 (Ad4) by DNA restriction endonuclease analysis during 1993-1995, when no epidemic outbreak of conjunctivitis occurred. METHODS: We evaluated 16 Ad4 strains from patients with acute viral conjunctivitis at four eye clinics in Sapporo, northern Japan. Seven strains were obtained during the period from December 1993 through March 1994 (first period). Nine strains were obtained during the period from March through May 1995 (second period). These strains were analyzed using DNA restriction endonucleases, TaqI and HinfI. RESULTS: The seven strains obtained during the first period showed identical DNA digestion patterns. The nine strains obtained during the second period showed two DNA digestion patterns using TaqI: five strains showed the same digestion pattern as that seen in the strains obtained during the first period, and four showed a different pattern. The genetic changes in Ad4 during 1993-1995 were less frequent than those reported previously during 1985-1989. CONCLUSION: It is suggested that the decrease in the incidence of infection in 1993-1995 was related to the decrease in the incidence of mutation in the Ad4 DNA. PMID- 11033122 TI - Prevalence of Chlamydia trachomatis pooled serotypes BDE and FGK in children with chronic follicular conjunctivitis. AB - PURPOSE: To evaluate type-specific antichlamydial antibody IgG in children with chronic follicular conjunctivitis. METHODS: A total of 90 serum samples from juvenile patients with chronic follicular conjunctivitis were collected in the Southeastern Anatolian region of Turkey where trachoma is still endemic. These samples were investigated regarding Chlamydia trachomatis pooled serotype specific IgG by using the micro-immunofluorescence test. A titer of 1/32 or higher was considered positive. RESULTS: Specific IgG seropositivity to Chlamydia trachomatis titer was found in 33 (36.1%) of the 90 subjects. A higher titer was observed frequently in the serotypes pooled in BDE (21 subjects), CHIJ (10 subjects), and FGK (2 subjects), respectively. CONCLUSIONS: The presence of antichlamydial antibodies in blood should always be interpreted in accordance with the history of the patient, the clinical picture, and the course of the disease. In the diagnosis of Chlamydia trachomatis infections in a patient with chronic follicular conjunctivitis, not only genus-specific antibodies but also the presence of the subspeciespecific antibodies should be investigated. PMID- 11033123 TI - Glucose transporter 1 expression in corneal wound repair under high serum glucose level. AB - PURPOSE: To determine glucose transporter (GLUT) 1 mRNA and protein expression during corneal epithelial wound healing in diabetic rat. METHODS: Diabetes mellitus was induced by intraperitoneal injection of streptozotocin. At 10 days after injection, unilateral 3-mm epithelial debridement was carried out in the central cornea. At 2, 4, 6, and 24 hours after wounding, whole corneal epithelium was collected and GLUT1 protein and mRNA levels were determined by Western blotting and reverse transcription-polymerase chain reaction, respectively. Sugar content in collected samples was measured by the Anthrone reaction. Normal rats were used as controls. RESULTS: Glucose transporter 1 protein and mRNA levels in unwounded cornea were similarly low in the diabetic and control groups. Healing of corneal wounds was slower in diabetic rats than in controls. After wounding, GLUT1 mRNA and protein expression in both groups were similarly enhanced compared to unwounded epithelium. Sugar content at all time points did not show significant alteration in any group, although in diabetic rats it was significantly higher than in controls throughout the time course. CONCLUSION: Glucose transporter 1 expression in diabetic rat cornea showed little difference from that in normal rat cornea, suggesting minimal influence of GLUT1 on the delayed healing of diabetic corneal wounds. PMID- 11033124 TI - The effect of sodium hyaluronate on the expression of gelatinases in rabbit corneal epithelial wound healing. AB - PURPOSE: We investigated the effect of sodium hyaluronate (Na-HA) on the expression of gelatinases in a rabbit model with corneal epithelial defects. METHODS: Topical administration of Na-HA or phosphate-buffered saline (PBS) was carried out in the experimental eyes after surgical removal of the corneal epithelium. At 0, 6, 24, 48, and 72 hours after wounding, mRNA expression of 72 kDa type gelatinase (MMP-2), 92 kDa type gelatinase (MMP-9), and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) were analyzed by reverse transcription polymerase chain reaction in those corneas. In addition, gelatinolytic activities were investigated using gelatin zymography. RESULTS: The levels of constitutive expression of MMP-2 and TIMP-1 mRNA persisted, whereas MMP-9 mRNA in the PBS treated side was expressed temporarily after surgical removal. In the Na-HA treated side, at 6 hours after wounding, a much higher expression of MMP-9 mRNA was reproducibly observed compared with that in the PBS-treated side. In zymography, the levels of gelatinolytic activity corresponding to proMMP-9 were significantly higher in the Na-HA-treated side than in the PBS-treated side at 6 hours after wounding. CONCLUSIONS: These results suggest a novel participation of Na-HA in the expression of MMP-9 in rabbit corneal epithelial wound healing. PMID- 11033125 TI - Immunohistochemical study of localization of extracellular matrix after holmium YAG laser irradiation in rat cornea. AB - PURPOSE: To better understand the corneal responses to holmium YAG (Ho:YAG) laser irradiation, we used immunofluorescent microscopy to examine changes in the localization of the extracellular matrix components, which play important roles in the maintenance of corneal morphology and functions. METHODS: Rats were irradiated with a Ho:YAG laser. On days 1, 3, and 7 after irradiation, the eyes were enucleated and frozen. Cryosections were made with a cryostat and were stained with antibodies against type I collagen, fibronectin, type IV collagen, or laminin for immunohistochemical study. RESULTS: One day after Ho:YAG laser irradiation, contraction of the stromal collagen fibrils was observed. Keratocytes could not be observed at the irradiated stromal region on day 1 after irradiation. One week later, however, keratocytes returned to the irradiated area. Although the stromal collagen fibrils had contracted, they were stained by an antibody against type I collagen. Dense fluorescence for fibronectin was observed at the margin of the stromal acellular zone. Both laminin and type IV collagen were observed at the basement membrane under the corneal epithelium, regardless of whether or not the corneas had been irradiated. CONCLUSION: These results suggest that Ho:YAG laser irradiation might be useful for the collagen contraction of stroma, without causing serious damage to the corneal epithelium and the basement membrane. PMID- 11033126 TI - Effect of decreased retinal illumination on frequency doubling technology. AB - PURPOSE: To investigate the effect of changes in retinal illumination on Frequency Doubling Technology (FDT). METHODS: Five eyes, of 5 adults who were free from identifiable ocular pathology, were examined using the Snellen chart and the Pelli-Robson chart, conventional automated perimetry, and the full threshold N-30 program of FDT. Each test was performed with and without a 0.9, 1.5, or 2.4 log unit neutral-density (ND) filter placed before the eye. Furthermore, the influence of pupil diameter on FDT test results was compared after treatment with pilocarpine or cyclopentolate with the influence of ND filters. RESULTS: All tests showed a decrease in sensitivity with decreasing retinal illumination. Frequency Doubling Technology showed an especially pronounced and significant decrease in sensitivity. The maximum mean threshold difference in FDT results with ND filter was 31.2 dB while that with the Humphrey Field Analyzer and the Pelli-Robson chart were 13.3 dB and 0.66 log contrast, respectively. The mydriatic state of the pupil increased the sensitivity of FDT and the miotic state decreased it to about the same extent as the the 0.9 ND filter. CONCLUSION: The change in retinal illumination has more impact on FDT than on spatial contrast tests and conventional automated perimetry. It is important to take this into account in evaluating FDT results. PMID- 11033127 TI - Role of Pseudomonas aeruginosa culture filtrates in the association, invasion, and cytotoxicity against cloned cells from murine corneal epithelium and KB cells. AB - PURPOSE: To clarify the effect of Pseudomonas aeruginosa culture filtrates on the association with, invasion into, and cytotoxicity against cloned cells from murine corneal epithelial cells and KB cells. METHODS: Simian virus 40 transformed murine corneal epithelial (MCE) cells were established. Murine corneal epithelial cells and KB cells were infected with a protease-positive strain, IID1117 (Pa IID1117), and a protease-negative strain, IID1130 (Pa IID1130) of P. aeruginosa, and then tested for association and invasion of Pa IID1117. The cytotoxicity test was performed by incubating the cells with culture filtrate. RESULTS: Association of Pa IID1117 with KB cells pretreated with Pa IID1130 was significantly promoted. After pretreatment with culture filtrate, invasion was more effective into MCE cells than into KB cells. When infecting bacteria (Pa IID1117) were pretreated with protease inhibitor, invasion of the bacteria into MCE cells and KB cells clearly decreased. The cellular damage induced by the culture filtrate of Pa IID1130 was greater than the damage by that of Pa IID1117. CONCLUSION: These results suggest that association of P. aeruginosa with MCE cells and KB cells was influenced by the culture filtrates other than proteases, and that invasion of P. aeruginosa into MCE cells and KB cells was promoted by protease. PMID- 11033128 TI - Immunohistochemical detection of retinal cones in monkey retina: light and electron microscopic study. AB - PURPOSE: Retinal cones have wave-length-specific visual pigments. To identify subclasses of cones, opsin-specific antibodies were previously established for the immunohistochemical studies of frozen sections. In this study, we produced retinal cone antibodies and examined (1) the specificity of these antibodies with Western blot analysis, (2) the application of these antibodies to paraffin embedded monkey retinal sections, and (3) the use of these antibodies in light and electron microscopic immunohistochemical analyses of the localization of retinal cones. METHODS: The N-terminal peptide of blue opsin, and the C-terminal peptide of green/red opsin were used as immunogens in New Zealand White rabbits. Immunohistochemical staining was performed using the ABC method and immunogold method. As antigen retrieval treatment, paraffin-embedded cynomolgus monkey retinas were subjected to enzyme and microwave treatment. RESULTS: Both anti red/green and anti-blue cone opsin antibodies detected 40 kDa native cone opsins in crude retina extract. The red/green and blue cone opsin immunostaining after trypsin treatment revealed that a positive signal was observed in the cone outer segment. Immunogold labeling also showed that gold particles were concentrated on the cone outer segment. CONCLUSION: In the antigen retrieval method, trypsin treatment is the appropriate method to obtain optimal staining of paraffin sections of retina. Using this method, retinal cone in conserved old paraffin sections can be identified immunohistochemically. PMID- 11033129 TI - Combined medical and surgical treatment of severe vernal keratoconjunctivitis. AB - PURPOSE: We report the efficacy of an alternative method of treatment for vernal keratoconjunctivitis (VKC) that consists of excision of the palpebral conjunctiva followed by supratarsal injection of corticosteroid and five times daily topical application of 0.05% cyclosporine A (CsA) and cromolyn sodium. METHODS: We evaluated 10 patients with severe treat-resistant VKC with corneal complications. The patients were evaluated for symptoms and for signs, including conjunctival changes, corneal limbal infiltrates, vascularization, reduction of epitheliopathy, meibomitis, visual acuity, intraocular pressure, and pathologic evaluation, before and after treatment. RESULTS: All patients showed marked improvement after 2 weeks of treatment. The symptoms (P <.01), signs (P <.02), and the visual acuity of all patients (P <.01) had significantly improved following treatment. Histological examination showed significant inflammatory cell decreases 4 weeks after surgery (P <. 05). CONCLUSION: Surgery plus topical drug therapy may be useful in treating patients with very severe VKC. PMID- 11033130 TI - The characteristics of static visual fields in children with psychogenic visual disturbances. AB - PURPOSE: It is well-known that patients with psychogenic visual disturbances (PVD) exhibit characteristic kinetic visual fields. Even when the kinetic fields are normalized, the static fields of PVD children frequently remain abnormal. To verify this finding, we performed static perimetry on those children whose kinetic fields were initially normal or which normalized during the follow-up period, and compared the results with those of children with psychosomatic disorders (PSD) and normal children. METHODS: We examined 9 PVD children (17 eyes), 16 PSD children (32 eyes), and 16 normal children (16 eyes). Program 30-2 or 24-2 of the Humphrey Field Analyzer was used in the examinations on all subjects. RESULTS: The average mean deviation (MD) of the PVD group was significantly lower than that of the other groups (P <. 01). False negative errors and short-term fluctuations were significantly higher in the PVD group than in the other groups (P <. 05). CONCLUSION: Although PVD and PSD children possess a similar underlying psychological dysfunction, their performances in visual field testing proved to be quite different. In the PVD group, even when kinetic fields were normal, functional visual field loss in the static fields was common and had characteristic response properties. PMID- 11033131 TI - A case of noninflammatory corneal edema following anterior-posterior radial keratotomy (Sato's operation) successfully treated by topical corticosteroid. AB - BACKGROUND: Corticosteroids seem to affect the functions of corneal endothelial cells directly in addition to their anti-inflammatory effect. CASE: A 69-year-old man presented with blurred vision in his left eye and decreased visual acuity in both eyes. He had received anterior-posterior radial keratotomy for myopia in both eyes 40 years earlier. OBSERVATIONS: Both eyes showed mild corneal stromal edema in the pupil area. We diagnosed the condition as corneal edema secondary to cell loss and dysfunction of corneal endothelium after surgery. One month later, the stromal edema was further advanced in the left eye. Corneal thickness had increased from the initial value of 619 microm to 631 microm, and corrected visual acuity had decreased from 0.5 to 0.06. There were no signs suggesting inflammatory reaction. The left eye was then treated with topical corticosteroid eyedrops and subconjunctival corticosteroid injection. Two months later, corneal thickness had decreased to 546 microm and visual acuity had improved to 0.5. CONCLUSIONS: This case shows that topical corticosteroids may be effective in treating corneal edema by producing an anti-inflammatory effect and possibly activating the endothelium. PMID- 11033132 TI - Mitomycin C primary trabeculectomy with releasable sutures in primary glaucoma. AB - PURPOSE: To evaluate the effects of mitomycin C and a releasable suture technique on outcomes of primary trabeculectomy in primary glaucoma patients. METHODS: A prospective analysis of patients who underwent primary trabeculectomy with a mitomycin C concentration of 0.2 mg/mL for 2 minutes. For closing the scleral flap, releasable sutures were used in 18 patients (17 eyes), Group 1, or permanent sutures in 18 patients (20 eyes), Group 2. Clinical outcome factors including postoperative intraocular pressure (IOP), visual acuity, and incidence of complications were determined. RESULTS: The mean follow-up periods were 8.1 +/ 1.3 months in Group 1 and 8.3 +/- 1.3 months in Group 2. The postoperative reduction in IOP was highly significant (P <.0001) at all time intervals in both groups. In all measurement of IOP before the second week, mean IOP in Group 2 was found significantly lower than the mean IOP in Group 1 (P =.01). No statistically significant differences were found between the groups at later mean IOP measurements. At the last visit, the complete success rate was 88.8% in Group 1 and 85.0% in Group 2. No serious complications such as hypotonous maculopathy were observed in any patient. CONCLUSION: Primary trabeculectomy with mitomycin C in eyes with primary glaucoma showed effective IOP pressure reduction. There were no cases of serious complications. In the early postoperative period IOP was controlled better in the releasable suture group. PMID- 11033133 TI - Long-term outcome after radiation therapy for subfoveal choroidal neovascularization associated with age-related macular degeneration. AB - PURPOSE: The purpose of this study was to investigate the long-term effect of low dose radiation therapy on subfoveal choroidal neovascularization associated with age-related macular degeneration. METHODS: The clinical course and visual outcome were compared retrospectively among two treated groups and a control group; 15 patients (15 eyes) received 10 Gy, another 15 patients (15 eyes) received 20 Gy. The control group consisted of 15 patients (15 eyes) without treatment. All patients were followed up for at least 18 months, and most were followed up for 3 years. The macula was irradiated with either 10 Gy in 5 fractions or with 20 Gy in 10 fractions after computed tomography (CT) simulation enabled real-time treatment planning from multiple CT slices. RESULTS: During the 3 years of follow up, the lesions became better in 5 eyes, unchanged in 1, and worse in 9 with 10 Gy radiation; better in 7 eyes, unchanged in 1, and worse in 7 eyes with 20 Gy; and better in 1 eye and worse in 14 with no treatment. The difference between the groups treated with radiation and the control was statistically significant (P <.05). Visual acuity was also significantly better in the group receiving 20 Gy than in the control group up to 2 years after radiation (P <.01). CONCLUSION: Radiation may extend the period of good visual function substantially by reducing subfoveal choroidal neovascularization activity. PMID- 11033134 TI - Two-year follow-up study comparing primary vitrectomy with scleral buckling for macula-off rhegmatogenous retinal detachment. AB - PURPOSE: To compare the anatomic and visual outcomes achieved by scleral buckling and primary vitrectomy for the repair of macula-off rhegmatogenous retinal detachment. METHODS: The records were reviewed for a consecutive series of 167 patients (167 eyes) who were initially treated with scleral buckling or pars plana vitrectomy for primary macula-off retinal detachment. Patients were treated between January 1993 and December 1996. After adjustments for preoperative characteristics, data from 102 cases (55 scleral buckle cases and 47 primary vitrectomy cases) were used for the final comparison. There had been a minimum follow-up period of 24 months. RESULTS: No significant differences in single procedure reattachment incidence (91%), final success incidence (100%) and incidence of postoperative proliferative vitreoretinopathy development (4%) were observed between the two treatment groups. Preoperative visual acuity, preoperative intraocular pressure, and duration of macular detachment were the three best predictors of postoperative visual recovery in both groups. Favorable overall visual recovery was obtained postoperatively, with no significant differences between the two groups throughout the follow-up period. However, in the eyes with poor preoperative visual acuity (<0.1), ocular hypotony (intraocular pressure <7 mm Hg), or prolonged macular detachment (more than 7 days), visual recovery in the primary vitrectomy group was significantly better (P <.05) than in the scleral buckle group from the first postoperative month. CONCLUSION: Both procedures achieved favorable anatomic and visual outcomes in the majority of patients with primary macula-off retinal detachment. Primary vitrectomy may be more effective than scleral buckling for achieving early visual rehabilitation in cases complicated by poor preoperative vision, ocular hypotony, and prolonged macular detachment. PMID- 11033135 TI - Optic neuritis in herpes zoster ophthalmicus. AB - BACKGROUND: Optic neuritis in herpes zoster ophthalmicus (HZO) has been reported rarely. We report two cases of HZO optic neuritis with detailed magnetic resonance imaging study and treatment responses. CASES: One patient presented with anterior optic nerve involvement, and the second presented with retrobulbar optic neuritis. Contrast enhanced T(1)-weighted images were obtained in these 2 patients. Intravenous acyclovir and oral prednisolone were given simultaneously. OBSERVATIONS: Magnetic resonance imaging revealed peripheral enhancement of the optic nerve sheath complex on T(1)-weighted scan. Both patients recovered their vision within 3 months following the start of treatment. CONCLUSIONS: Magnetic resonance imaging is helpful for the diagnosis of HZO optic neuritis. Systemic acyclovir and steroid are effective in the treatment of HZO optic neuritis. PMID- 11033136 TI - Myectomy of lateral rectus muscle for third nerve palsy. AB - PURPOSE: To introduce myectomy of the lateral rectus muscle for correcting exotropia in patients with third nerve palsy. METHODS: The lateral rectus muscle of the paretic eye was myectomized without suturing it to the globe. This was combined with a medial rectus muscle resection and a contralateral lateral rectus muscle recession. Magnetic resonance imaging was performed to observe the re attachment of the lateral rectus muscle to the globe. RESULTS: The patient was able to fuse in the primary position without any noticeable limitation in abduction. Magnetic resonance imaging showed that the lateral rectus muscle was attached to the globe through fibrous tissue. CONCLUSION: Myectomy of the lateral rectus muscle is an effective and simple procedure to accomplish a super-maximal weakening effect of abduction in patients with complete third nerve palsy. PMID- 11033137 TI - Acute conjunctivitis associated with biofilm formation on a punctal plug. AB - BACKGROUND: Punctal plugs are used for the treatment of tear-deficient type dry eye. We recently examined a case of acute conjunctivitis associated with bacterial biofilm formation on a punctal plug. CASE: A 63-year-old woman diagnosed as having tear-deficient type dry eye came to our hospital with a complaint of soreness in her right eye. Punctal plugs had been inserted into this eye 5 1/2 months previously. On the day of her visit, she presented with acute conjunctivitis. OBSERVATIONS: In biomicroscopical examination, the top of the punctal plug was seen to be covered with a whitish soft material. Microbiological analysis performed on a part of this material was positive for Staphylococcus haemolyticus and Candida tropicalis. Scanning electron microscopy of the removed punctal plug revealed widespread bacterial colonization embedded within an extensive extracellular matrix. Treatment consisted of the replacement of the plug, and administration of a combination of antibacterial eyedrops and preservative-free artificial solution. As a result, the acute conjunctivitis cleared up within 1 month. CONCLUSIONS: This case suggests that a punctal plug poses a potential risk of causing the formation of bacterial biofilm. In such a case, replacement of the plug and/or removal of the accumulated materials should be considered. PMID- 11033138 TI - Acute retinal necrosis following contralateral herpes zoster ophthalmicus. AB - BACKGROUND: A case report of contralateral acute retinal necrosis (ARN) following herpes zoster ophthalmicus. CASE: A 61-year-old male patient developed iridocyclitis and well-demarcated creamy-white retinal lesions at the nasal periphery in the right eye 1 month after herpes zoster ophthalmicus in the left eye. The patient had undergone surgery for primary lung cancer, and had subsequent intracranial metastasis of the tumor. OBSERVATIONS: The clinical diagnosis of ARN was supported by polymerase chain reaction investigation of the aqueous humor resulting in positive for varicella-zoster virus. Retinal lesions disappeared after systemic treatment with acyclovir, corticosteroids, and acetylsalicylate. No retinal detachment developed. CONCLUSIONS: We propose a careful ophthalmic follow-up for herpes zoster ophthalmicus patients because of the possibility of acute retinal necrosis developing in the contralateral eye. PMID- 11033139 TI - Amiodarone-related optic neuropathy. AB - BACKGROUND: To evaluate a case of atypical optic neuropathy that presented with blurred vision following the use of an antiarrythmic agent. CASE: Record of the patient was reviewed to determine the etiology of his optic neuropathy. OBSERVATIONS: Ophthalmological examination revealed unilateral optic disc edema with relatively well-preserved visual acuity. In routine tests, results of complete blood count, erythrocyte sedimentation rate, liver and kidney function tests, chest x-ray, Goldmann visual field examination, and brain computed tomography scan were normal. Orbital ultrasonography revealed optic disc edema with prominent optic nerve head and without orbital pathology. CONCLUSIONS: Systemic history and drug intake should be investigated in every patient with optic disc edema. Discontinuation of the medication can prevent further optic nerve damage or involvement of the other eye. PMID- 11033140 TI - Identification of a Free Non-tryptophan Fluorophore in Water-soluble Fraction of Human Brunescent Cataractous Lens Nucleus. AB - Purpose: We previously reported that a unique free fluorophore (Fl-Glc), presumably a beta-glucoside, is particularly abundant in human brunescent cataractous lens nuclei. Our preliminary experiments indicated that incubation of low-molecular weight (MW) fraction of non-brunescent lens nuclei causes an increase in a particular fluorophore (Fl-X). This study was undertaken to compare the Fl-Glc with the Fl-X and subsequently to identify the Fl-X.Methods: Experiment (1) The purified Fl-X and its beta-glucosidase digest (aglycon) were compared with the Fl-Glc and its aglycon, respectively, by high-performance liquid chromatography (HPLC). Experiment (2) i) The Fl-X and its aglycon were analysed by liquid chromatography/mass spectrometry (LC/MS). ii) Authentic xanthurenic acid was analysed by HPLC and LC/MS.Results: Experiment (1) The retention times of the Fl-X and the Fl-Glc exactly coincided. The fluorescence peaks of both disappeared after beta-glucosidase treatment. Experiment (2) i) LC/MS results suggested that the MWs of the Fl-X and its aglycon were 367 and 205, respectively. ii) HPLC and LC/MS results for xanthurenic acid (MW = 205) were exactly the same as those for the aglycon of the Fl-X.Conclusions: The Fl Glc and the Fl-X are identifical, and the Fl-X (= Fl-Glc) is a glucoside of xanthurenic acid. PMID- 11033141 TI - The Effect of Intraocular Lidocaine in White Rabbit Eyes. AB - Purpose: Recently, intraocular lidocaine anesthesia has been used in cataract surgery. We studied the toxicity of intraocular unpreserved lidocaine for corneal endothelial cell and retina using Japanese white rabbits.Methods: The rabbits were divided into two groups. One group was injected intracamerally and the other was injected intravitreally with 0.2 ml of unpreserved lidocaine of 0%, 0.02%, 0.2%, or 2% concentration. The number of corneal endothelial cells was measured 1 week after the injection. After measurements, the rabbit corneas were studied histologically. The retina was examined by electroretinogram prior to initial injection through 1 week after the injection.Results: There was no significant change in number of corneal endothelial cells after injection of 0.2% lidocaine. However, histological abnormality was seen in corneal endothelial cells after 2% lidocaine injection. There was also significant change in electroretinogram with 2% lidocaine injection. No histological abnormality was seen in the retina 1 week after the injection.Conclusion: The rabbit cornea and retina manifested no serious changes after the injection of lidocaine at less than 0.2% concentration functionally and histologically. PMID- 11033142 TI - Immunohistochemical Study of Apoptosis of Lens Epithelial Cells in Human and Diabetic Rat Cataracts. AB - Purpose: An immunohistochemical evalution of lens epithelial cell apoptosis.Methods: We performed terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays on capsulotomy specimens (68 eyes in 53 patients) from patients who had undergone cataract surgery and an epithelium of diabetic cataracts in rats (144 eyes in 72 rats). The animal model of diabetic cataracts was prepared by injection of streptozotocin in three-week old rats. The rats were also examined using the proliferating cell nuclear antigen (PCNA) immunohistochemical staining method.Results: Although some TUNEL-positive cells were detected in capsulotomy specimens, we recognized little correlation between its distribution and morphological classification of cataracts. In the animal model of diabetic cataracts, TUNEL-positive cells were seen around the region where epithelial cells had accumulated. In the accumulated region, PCNA labeled cells undergoing DNA synthesis were also detected.Conclusion: These results suggest the possibility that apoptosis occurs in human lens epithelial cells and apoptosis and proliferation may be induced by factors such as hyperglycemia. PMID- 11033143 TI - Effect of Carteolol Hydrochloride on Ocular Blood Flow Dynamics in Normal Human Eyes. AB - Purpose: The effect of topical 2% carteolol hydrochloride on the ocular blood flow dynamics in normal human eyes was studied.Materials and Methods: Ten healthy volunteers were studied using color Doppler imaging and scanning laser Doppler flowmetry. In the first experiments, one eye received 30 ul of 2% carteolol hydrochloride twice daily for 7 days and the fellow eye a placebo in a randomized masked manner. The blood flow velocity of the central retinal artery, the ophthalmic artery, and the short posterior ciliary artery, the blood flow volume of the peripapillary retina (PPR), intraocular pressure (IOP), ocular perfusion pressure (OPP) and pulse rate (PR) were measured before treatment, 2 hours after the initial instillation, and after the last instillation on the 7th day. In the control experiments, 30 ul of placebo was instilled in both eyes, and the above parameters were measured according to the same time schedule as in the first experiments.Results: In the carteolol-treated eyes, the systolic maximum (p = 0.012) and diastolic minimum (p = 0.019) blood flow velocities of the ophthalmic artery, and the blood flow volume of the superior (p = 0.003), central (p = 0.001) and inferior (p = 0.000) PPR showed significant increases in the first experiments compared with the control experiments. IOP and OPP in both eyes showed significant reduction in the first experiments compared with the control study.Conclusion: These results indicated that topical carteolol hydrochroride increases the blood flow of the intraorbital microcirculation and PPR. PMID- 11033144 TI - The Rayleigh Color Matches in Idiopathic Macular Holes Treated by Vitrectomy. AB - Purpose: The Rayleigh color matches were measured to investigate the optical density in the cone photopigment of the central retina in patients with macular holes that had closed after vitrectomy.Methods: The Rayleigh equation was measured with the IF-2 anomaloscope in 7 patients with macular holes in one eye, that had closed after vitrectomy and gained an improvement in visual acuity of 0.5 or better. Reductions in the pigment optical density of cone photoreceptors of the opposite eyes were estimated with Rayleigh equations obtained in this experiment.Result: The Rayleigh equations of the affected eyes were shifted toward protanomalous setting compared with that of the opposite eyes. The optical densities of cone photoreceptors of the affected eyes were lower than those in the opposite eyes, even in patients with visual acuity of 1.0 or better.Conclusion: These findings suggest that the recovery of the visual acuity may precede that of the optical density in cone photopigment of central retina. The Rayleigh equation is more sensitive than visual acuity for evaluating visual function after vitrectomy for macular holes. PMID- 11033145 TI - Multifocal Electroretinogram in Central Serous Chorioretinopathy. AB - Purpose: An assessment of retinal function topographically in the affected eyes with central serous chorioretinopathy (CSC).Participants and Method: We recorded multifocal electroretimogram (mERG) in 5 patients with unilateral CSC, and analyzed the topographical properties in the central visual field (rings #1 + 2).Results: mERG amplitudes in the CSC eyes were significantly reduced compared with the fellow eyes, for P 1-N 1 (58.5 +/- 23.5%, p < 0.05) and for P 1-N 2 (47.5 +/- 15.0%, p < 0.05). mERG latencies in the CSC eyes were significantly increased compared with those in the fellow eyes, for N 1 (by 9.0 +/- 8.1%, p < 0.05) and for P 1 (by 8.4 +/- 7.0%, p < 0.05). mERG in the CSC eyes gradually recovered in the follow-up period. However, the responses did not recover to normal value during follow-up, even when the subretinal fluid disappeared ophthalmoscopically.Conclusions: These results show that a topographical analysis of the mERG is useful for clinical observation of CSC. PMID- 11033146 TI - The Difference in Saccadic Parameters among Several Visually Guided Tasks Saccadic Reaction Time. AB - Purpose: The saccadic reaction time (SRT) was measured as a saccadic movement parameter using visually guided tasks.Subjects and Method: The SRT was measured for nine healthy adult subjects aged from 28 to 49 years. The visually guided tasks for saccadic movement induction were simultaneous, gap, overlap, delayed memory, and anti-SM tasks.Results: The median of the SRT (msec) for each task in the actual measurements was as follows: 216 for the simultaneous task, 180 for the gap task, 240 for the overlap task, 234 for the delayed-memory task, and 292 for the anti-SM task. The SRT distribution of each subject exhibited two peaks for the gap task but mostly one peak for each of the other tasks. The first peak, observed in the SRT distribution for the gap task was an eysress saccade.Conclusion: The SRT for the gap task was the shortest and the SRT got longer in the order of the simultaneous task, and the overlap task, and the anti SM task. Some subjects easily showed an express saccade in the gap task, but others did not. PMID- 11033147 TI - Vitrectomy for the Treatment of Expulsive Hemorrhage. AB - Purpose: An evaluation of surgical outcome of vitrectomy in the treatment of expulsive hemorrhage associated with intraocular surgery.Methods: We reviewed 12 eyes from 12 patients with expulsive hemorrhage, occurring after or during cataract extraction (4 eyes), phacoemulsification (2 eyes), glaucoma filtering surgery (4 eyes), or vitrectomy (2 eyes). Mean follow-up period was 21 months.Results: The retina was reattached in 6 eyes (50%) after the initial surgery and ultimately in 9 eyes (75%). Three eyes, which failed to achieve retinal reattachment, resulted in phthisis bulbi. Final visual acuity was 0.1 or better in 4 eyes and 0.01 to 0.09 in 4 eyes. The incidence of expulsive hemorrhage was 0% for cataract surgery, 0.57% for trabeculectomy, and 0.09% for vitrectomy at Toho University Sakura Hospital.Conclusion: In the treatment of expulsive hemorrhage, vitrectomy is an effective surgical procedure to improve the visual function. PMID- 11033148 TI - Histopathological Features of the Trabecular Meshwork in a Case of Presumed Bilateral Chandler Syndrome. AB - Background: We describe a patient who had no-table decrease in the number of corneal endothelial cells in both eyes and developed open angle glaucoma without evident iris atrophy and peripheral anterior synechia.Case: A 74-year-old woman. The trabeculectomized angle tissue and iris tissue of her left eye were observed under light and transmission electron microscopy. From Schwalbe's line to the anterior chamber angle, degenerated endothelium-like cells were observed and overgrowth of layers of collagen fiber with varied cycles and basement membrane like material were noticed. In addition, the morphology of intra-trabecular space was atrophic and occlusive with marked degeneration and exfoliation of endothelial cells in the trabecular space. On the side of the anterior chamber, degenerated endothelium-like cells were observed, the morphology of which was considered to result from abnormal metabolic function in corneal endothelium. Overgrowth of basement membrane-like material onto iridic tissue was not observed.Conclusion: Although slit lamp examination revealed no obvious abnormality, the result of histological examination suggested that this was a case of Chandler's syndrome in a broad sense or was in the early stage of this disease. We also discussed differential diagnosis from other diseases. PMID- 11033149 TI - Twenty-three Cases of Primary Cornea Guttata. AB - Purpose: An evaluation of the clinical aspects of patients with primary cornea guttata and the morphology of their corneal endothelial cells.Methods: Twenty three patients who visited Kanazawa Medical University Hospital or related hospitals in Ishikawa or Fukui prefectures and were diagnosed as having primary cornea guttata by slit lamp microscopy and contact specular microscopy underwent an analysis of corneal endothelial cells as well as their clinical status.Results: In 23 cases, the number of men and young people under 30 years of age was low, with 4 and 2 cases respectively. There was no visual impairment due to cornea guttata. In the specular microscopic findings, the size and density of dark areas varied according to each case and there was asymmetry in some cases. The parameters of the endothelial cells, comprising mean cell area, hexagonality, and the coefficient of variation, were almost within normal limits except for one eye of a 79-year-old man who was speculated to be at an early stage of Fuch's endothelial corneal dystrophy. Three cases had received cataract surgery, but showed no statistical changes in endothelial morphology before or after the surgery.Conclusion: Cornea guttata is thought to be a disease which is not uncommon in Japanese people and which does not bring about visual impairment or abnormalities in corneal endothelial cell parameters except for a small number of patients. Such cases with remarkably decreased cell densities should be considered to be at an early stage of Fuch's dystrophy. PMID- 11033150 TI - A Case of Reversed Ophthalmic Artery Flow without Occlusion of the Internal Carotid Artery. AB - Background: We report a case of reversed ophthalmic artery (OA) flow showing neither occlusion of the internal carotid artery (ICA) nor ophthalmic symptoms.Case: An 82-year-old man had transient blindness in both eyes. After left carotid endarterectomy, ophthalmic symptoms improved in the right and left eyes. Afterwards, he had a relapse in the left eye.Results: There was no unusual impression in the ophthalmic examinations, but color Doppler imaging showed reduced flow in the left OA, reversal of flow direction in the right OA. The central retinal artery (CRA) and short posterior ciliary arteries (SPCA) were not detected in the left eye but were detected in the right eye. Digital subtraction angiography demonstrated that the right OA was not contrasted with the right ICA, despite there being no stenosis or occlusion of the right ICA.Conclusions: We suggest that carotid surgery may affect the ocular circulation of the opposite side, and reversed OA flow as a collateral pathway from the external carotid artery may occur in the absence of carotid artery stenosis. Good circulation of the CRA and SPCA may preclude manifestation of ophthalmic symptoms even if the OA flow is reversed. PMID- 11033151 TI - Optic Disc Blood Flow Measured by Scanning Laser-Doppler Flowmetry Using a New Analysis Program. AB - Purpose: Using a new analysis program for scanning laser-Doppler flowmetry (SLDF) by a Heidelberg retina flowmeter (HRF), we studied the relation between flow and visual field or disc morphology.Subjects and Methods: In 42 eyes of 21 patients with normal tension glaucoma (NTG) the mean-flow of the HRF blood flow parameters at the disc rim was measured and analyzed by a new analysis program for perfusion maps (the SLDF analysis tool), to minimize the influence of large vessels or/and artifacts caused by small eye movements. We investigated whether difference of the mean-flow between a pair of eyes had any relation to differences between a pair of eyes in visual field indices and those in disc morphological measurements of the Heidelberg retina tomograph.Results: We found statistically significant correlations between the mean-flow and optic disc parameters (Disk Area, Cup Area, Height Variation Contour, Cup Volume, Rim Volume, Mean RNFL Thickness). We found no statistically significant correlations between the mean-flow and visual field parameters (mean deviation, corrected pattern standard deviation).Conclusion: The results suggested that eyes with less flow in the optic disc rim have more advanced glaucomatous morphological changes. PMID- 11033152 TI - Thickness of Retinal Nerve Fiber Layer Decreases after Vitreous Surgery for Idiopathic Macular Hole. AB - Purpose: The alteration of the retinal nerve fiber thickness (NFLT) before and after vitreous surgery for a macular hole in cases with or without visual field defects was investigated.Methods: The NFLT of 23 eyes with idiopathic macular hole was measured with scanning laser polarimeter up to 12 months after surgery. The NFLT was divided into superior, inferior, nasal, and temporal quadrants. The mean NFLT of each quadrant was analyzed statistically and compared to the results of visual field tests.Results: In all cases, the NFLT decreased significantly up to 3 months after surgery, but increased gradually up to 12 months. In cases with visual field loss, the thickness of the nerve fiber layer that corresponds to the visual field defect diminished strongly, in contrast with other quadrants. Even in cases without visual field loss, the NFLT decreased significantly a month after surgery, especially in the nasal and inferior quadrants.Conclusion: This study demonstrates that the thickness of retinal nerve fiber layer decreases at least one month after surgery, even in cases without visual field defect. This may suggest the existence of a subclinical complication of vitreous surgery. PMID- 11033153 TI - Electrophysiological Evaluation of the Protective Effect of Dimethylthiourea against Retinal Photic Injury. AB - Purpose: The Protective Effect of Dimethylthiourea (DMTU) against photic injury of the retina was evaluated by electroretinogram (ERG).Methods: In the DMTU treated group, 250, 500, or 750 mg/kg DMTU was administered intraperitoneally to albino rabbits at 24 hours and immediately before starting light exposure to the eye. In the control group, physiological saline was injected intraperitoneally instead of DMTU. Preservation rates of ERG a-, b- and c-wave amplitudes were defined as the percentages of the post-photic injury values to the pre-photic injury ones, and were compared between the control and the DMTU-treated groups.Results: In 750 mg/kg DMTU-treated group, the preservation rate of the a wave was significantly higher than that in the control group 24 hours after the photic injury. While the preservation rate of the c-wave was remarkably low in the control group 24 hours after the photic injury, it was significantly higher in the 500 and 750 mg/kg DMTU-treated groups. Furthermore, better preservation rates of the c-wave were noted with higher doses of DMTU.Conclusion: These results suggest that DMTU protects against photic injury of the retinal pigment epithelium and photoreceptors. PMID- 11033154 TI - Clinical Results of Selective Laser Trabeculoplasty. AB - Purpose: Selective laser trabeculoplasty (SLT) is a new technique aimed to developed to impact pigmented trabecular cells selectively. Compared with ordinary argon laser trabeculoplasty, it is expected to have fewer complications with more efficacy for open-angle glaucoma. In this study we performed SLT on 17 eyes of 10 patients with primary open-angle glaucoma and 1 eye with capsular glaucoma.Methods: Follow-up period was up to 10 months. Average energy irrachieted was 28.14 mJ (0.47 mJ x 59 spots) against pigmented trabecular band over the half circumference of anterior chamber angle.Results: Preoperative mean intraocular pressure (IOP) was 22.8 mmHg and postoperative mean IOP was decreased significantly to 8.6, 17.3, and 16.1 mmHg at 1, 3, and 6 months after treatment, respectively. The average maximum IOP reduction was 8.8 (3 approximately 1 8) mmHg after SLT. Among 11 eyes showing transient IOP elevation, 6 eyes had an elevation of more than 6 mmHg. No remarkable postoperative complications were noted.Conclusion: SLT is a safe and effective modality for the treatment of open angle glaucoma such as primary open-angle glaucoma (POAG) and capsular glaucoma. PMID- 11033155 TI - Sinusotomy with Intraoperative Mitomycin C Administration in Selected Eyes with Advanced Glaucomatous Damage. AB - Purpose: The surgical outcome of sinusotomy with intra-operative mitomycin C administration (0.04%, 3 minutes) in a total of 17 selected glaucoma eyes from 14 patients with advanced glaucomatous damage was examined. Selection criteria were (1) an eye with visual field defect threatening the central fixation, while the central visual acuity of the contralateral eye was already impaired; and/or (2) an aphakic or pseudophakic eye with advanced glaucomatous damage in a patient of advanced age.Materials and Methods: Patient's age, preoperative mean deviation (Humphrey 30-2 program) and intraocular pressure (IOP) averaged 63.5 +/- 11.1 (mean +/- standard deviation) years, -18.6 +/- 6.7 (mean +/- standard deviation) dB, and 20.7 +/- 4.1 (mean +/- standard deviation) mmHg, respectively.Results: No postoperative hypotony (< 5 mmHg) or deterioration of visual acuity equal to or more than 2 lines of Ishihara Visual Acuity Chart were encountered and the mean postoperative IOP ranged between 11 and 13 mmHg up to 18 months postoperatively with less medication than preoperatively. The success was defined as IOP /=7 cm, or the patient had been operated. Hypothyroidism was less common if less than a half of the thyroid bed was irradiated. CONCLUSION: The detection of hypothyroidism clinically is difficult, and the rate of hypothyroidism warrants routine assessment of thyroid function after irradiation of laryngeal cancer. PMID- 11033193 TI - Multivariate analysis of pulmonary fibrosis after electron beam irradiation for postmastectomy chest wall and regional lymphatics: evidence for non-dosimetric factors. AB - BACKGROUND AND PURPOSE: To evaluate the factors associated with pulmonary fibrosis after postmastectomy electron beam irradiation of chest wall and regional lymphatics in patients with breast cancer. MATERIALS AND METHODS: From July 1987 through July 1994, 109 women with stage II and III breast cancer receiving modified radical mastectomies were managed by postoperative electron beam irradiation. Doses of 46 to 50.4 Gy were delivered to the chest wall covered with bolus, internal mammary nodes, supraclavicular nodes and axillary lymph nodes via 12 or 15 MeV single portal electron beam. Seventeen patients received additional 10-16 Gy surgical scar boost via 9 MeV electron beam. Comparison of pre-treatment and post-treatment chest X-ray films were used to monitor the development of pulmonary fibrosis. RESULTS: Only Grade 1 radiation-induced late pulmonary toxicity was noted in 33 patients (29%). Twenty-six patients (24%) developed pulmonary fibrosis under unbolused chest wall. Lung fibrosis under bolused chest wall was noted in 11 patients (10%). Statistical difference (P<0.01) was noted between the incidence of fibrosis in these two sites. In multivariate analysis of lung fibrosis under unbolus-covered chest wall, the independent prognostic factors are low body mass index (BMI) (P<0.01), tamoxifen taking (P=0.03), and no treatment interruption (P=0.03). No independent factor was associated with lung fibrosis under bolus-covered chest wall in multivariate analysis. CONCLUSIONS: In the analysis of pulmonary fibrosis induced by unbolused electron beam, BMI rather than body weight and body height is a strong prognostic factor. Tamoxifen and short overall time can predispose the development of lung fibrosis. PMID- 11033195 TI - Estimates of the theoretical maximum daily intake of erythorbic acid, gallates, butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) in Italy: a stepwise approach. AB - The three recent EU directives which fixed maximum permitted levels (MPL) for food additives for all member states also include the general obligation to establish national systems for monitoring the intake of these substances in order to evaluate their use safety. In this work, we considered additives with primary antioxidant technological function for which an acceptable daily intake (ADI) was established by the Scientific Committee for Food (SCF): gallates, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and erythorbic acid. The potential intake of these additives in Italy was estimated by means of a hierarchical approach using, step by step, more refined methods. The likelihood of the current ADI to be exceeded was very low for erythorbic acid, BHA and gallates. On the other hand, the theoretical maximum daily intake (TMDI) of BHT was above the current ADI. The three food categories found to be main potential sources of BHT were "pastry, cake and biscuits", "chewing gums" and "vegetables oils and margarine"; they overall contributed 74% of the TMDI. Actual use of BHT in these food categories is discussed, together with other aspects such as losses of this substance in the technological process and percentage of ingestion in the case of chewing gums. PMID- 11033196 TI - Inhibition of 1,2-dimethylhydrazine-induced oxidative DNA damage in rat colon mucosa by black tea complex polyphenols. AB - The effect of black tea polyphenols on 1,2-dimethylhydrazine (DMH)-induced oxidative DNA damage in rat colon mucosa has been investigated. Fischer 344 rats were treated orally with thearubigin (TR) or theafulvin (TFu) for 10 days (40 mg/kg), injected ip with DMH (20 mg/kg) or saline and sacrificed 24 hr after DMH administration. The levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) were measured in colonic mucosa DNA and expressed as a ratio relative to 2'-deoxyguanosine (2dG). Control rat mucosa had 8-OHdG values of 1.12 +/- 0.14/10(5) dG (mean +/- SEM, n=11), whereas DMH-treated rats significantly higher values (1.52 +/- 0.14/10(5) dG, n=26, P<0.05). Pretreatment of rats with TR had significantly inhibited DMH-induced oxidative DNA damage 0.99 +/- 0.09/10(5) dG, n=10, P<0.05) and a similar, although less marked, effect was observed with TFu (1.15 +/- 0.19/10(5), n=9, P=0.06). These findings confirm that DMH causes oxidative DNA damage in the colon mucosa of rats and demonstrate that this effect is prevented by the consumption of complex polyphenols from black tea. PMID- 11033197 TI - Inhibition of tert-butyl hydroperoxide-induced cell membrane bleb formation by alpha-tocopherol and glutathione. AB - Multiple bleb formation on cell membrane is common during cell death. The effects of alpha-tocopherol and glutathione (GSH) on tert-butyl hydroperoxide (TBH) induced membrane changes in rat hepatocytes were studied. Over 60 min of exposure, TBH (0.5-2.0 mM) caused a dose-dependent membrane blebbing. Cells pretreated with buthionine sulfoximine, a GSH synthesis inhibitor, had significantly greater blebbing and lactate dehydrogenase (LDH) leakage under 0.5 mM TBH treatment as compared to cells without pretreatment. However, the protective effect of GSH disappeared when the TBH concentration was increased to 2.0 mM. In the presence of alpha-tocopheryl succinate (TS) pretreatment, it was noted that bleb formation, expressed as the percentage of cells bearing blebs, the average bleb size, or the onset of blebbing, was partially suppressed even when TBH concentration was 2.0 mM. TBH-induced thiobarbituric acid reactive substances and LDH leakage were completely abolished by TS pretreatment. Accompanying bleb formation, membrane-insoluble actin was noted to decrease by immunoblot assay. The decrease in actin was also suppressed by TS. These results indicated that intracellular GSH and alpha-tocopherol status are important to the TBH-induced cell membrane abnormality. Furthermore, TS plays a defensive role against blebbing when GSH is exhausted by TBH. PMID- 11033198 TI - Toxicity assessment of fumonisins using the brine shrimp (Artemia salina) bioassay. AB - The Fusarium mycotoxins fumonisin B(1) (FB(1)) (1) and B(2) (FB(2)) (2), their hydrolysed analogues HFB(1) (3) and HFB(2) (4) and the recently discovered fumonisin derivatives N-palmitoyl-HFB(1) (5) and N-carboxymethyl-FB(1) (6) were compared for their toxicity in a short term bioassay using brine shrimp (Artemia salina). The brine shrimp were hatched in artificial sea water and exposed to the fumonisins in microwell plates with a mortality endpoint after 48 hours. LC(50) values were calculated after Probit transformation of the resulting data. Of the substances tested, fumonisin B(1) emerged to be the most toxic whereas its N carboxymethyl analogue was 100-fold less effective. The hydrolysed fumonisins showed a four- to sixfold reduced toxicity compared to FB(1). N-Palmitoyl-HFB(1) had a higher LC(50) value than its precursor HFB(1). The brine shrimp assay proved to be a convenient and rapid system for toxicity assessment of this group of mycotoxins. PMID- 11033199 TI - Developmental toxicity of Spinosad administered by gavage to CD rats and New Zealand white rabbits. AB - The insecticide Spinosad was administered by gavage to pregnant CD(R) rats at 0, 10, 50 or 200 mg/kg/day on gestation days (gd) 6-15 and to New Zealand White rabbits at 0, 2.5, 10 or 50 mg/kg/day on gd-7-19. Rats and rabbits were monitored for clinical signs of toxicity and body weight gains. At gd-21 (rats) or gd-28 (rabbits), maternal organ weights, reproductive parameters, fetal body weights, and fetal external, visceral and skeletal structures were evaluated. Rats given 200 mg/kg/day exhibited a 4% lower body weight on gd-12 and decreased body weight gains on gd-6-16 relative to controls. There was no maternal toxicity at 10 or 50 mg/kg/day, and no developmental toxicity in rats at any dose level. Rabbits given 50 mg/kg/day exhibited decreased feed consumption, reduced fecal output, body weight loss during the initial dosing period (gd-7-10) and a non-statistically significant decrease (31%) in body weight gain during the dosing period (gd-7 20). Two litters aborted due to maternal inanition. There were no maternal effects at lower doses, and no signs of developmental toxicity at any dose. Thus, the maternal no-observed-effect levels (NOEL) were 50 and 10 mg/kg/day in rats and rabbits, respectively, and the embryonal/fetal NOELs were 200 mg/kg/day in rats and 50 mg/kg/day in rabbits. PMID- 11033201 TI - Foreword PMID- 11033200 TI - Modulatory effects of a tannin fraction isolated from Terminalia arjuna on the genotoxicity of mutagens in Salmonella typhimurium. AB - A fraction isolated from Terminalia arjuna was studied for its antimutagenic effect against 4-nitro-o-phenylenediamine (NPD) in TA98, sodium azide in TA100 and 2-aminofluorene (2AF, S9-dependent), a promutagen, in both TA98 and TA 100 tester strains of Salmonella typhimurium using the Ames assay. The fraction inhibited the mutagenicity of 2AF very significantly in both strains while the revertant colonies induced by NPD and sodium azide were reduced moderately. 1H NMR, 13C-NMR, IR and UV-spectroscopic data of the fraction revealed it to be tannin in nature. PMID- 11033202 TI - Adaptive plasticity in vestibular influences on cardiovascular control. AB - Data collected in both human subjects and animal models indicate that the vestibular system influences the control of blood pressure. In animals, peripheral vestibular lesions diminish the capacity to rapidly and accurately make cardiovascular adjustments to changes in posture. Thus, one role of vestibulo-cardiovascular influences is to elicit changes in blood distribution in the body so that stable blood pressure is maintained during movement. However, deficits in correcting blood pressure following vestibular lesions diminish over time, and are less severe when non-labyrinthine sensory cues regarding body position in space are provided. These observations show that pathways that mediate vestibulo-sympathetic reflexes can be subject to plastic changes. This review considers the adaptive plasticity in cardiovascular responses elicited by the central vestibular system. Recent data indicate that the posterior cerebellar vermis may play an important role in adaptation of these responses, such that ablation of the posterior vermis impairs recovery of orthostatic tolerance following subsequent vestibular lesions. Furthermore, recent experiments suggest that non-labyrinthine inputs to the central vestibular system may be important in controlling blood pressure during movement, particularly following vestibular dysfunction. A number of sensory inputs appear to be integrated to produce cardiovascular adjustments during changes in posture. Although loss of any one of these inputs does not induce lability in blood pressure, it is likely that maximal blood pressure stability is achieved by the integration of a variety of sensory cues signaling body position in space. PMID- 11033203 TI - Patterning of sympathetic nerve activity in response to vestibular stimulation. AB - Growing evidence suggests a role for the vestibular system in regulation of autonomic outflow during postural adjustments. In the present paper we review evidence for the patterning of sympathetic nerve activity elicited by vestibular stimulation. In response to electrical activation of vestibular afferents, firing of sympathetic nerves located throughout the body is altered. However, activity of the renal nerve is most sensitive to vestibular inputs. In contrast, high intensity simultaneous activation of cutaneous and muscle inputs elicits equivalent changes in firing of the renal, superior mesenteric and lumbar colonic nerves. Responses of muscle vasoconstrictor (MVC) efferents to vestibular stimulation are either inhibitory (Type I) or are comprised of a combination of excitation and inhibition (Type II). Interestingly, single MVC units located in the hindlimb exhibited predominantly Type I responses while those located in the forelimb and face exhibited Type II responses. Furthermore, brachial and femoral arterial blood flows were dissociated in response to vestibular stimulation, such that brachial vascular resistance increased while femoral resistance decreased. These studies demonstrate that vestibulosympathetic reflexes are patterned according to both the anatomical location and innervation target of a particular sympathetic nerve, and can lead to distinct changes in local blood flow. PMID- 11033204 TI - Effects of caloric stimulation on respiratory frequency and heart rate and blood pressure variability. AB - Heart rate variability (HRV), blood pressure variability (BPV) and respiratory frequency were measured by power spectrum techniques in six normal humans (25-34 years old) and one labyrinthine-defective patient (33 years old) during cold (30 degrees ) vestibular caloric stimulation. Caloric stimuli were delivered intermittently for 2 min, under two different breathing conditions: (1) spontaneous breathing and (2) breathing paced with a metronome (0.25 Hz). During the spontaneous breathing condition, in the normal subjects, the caloric stimuli induced a significant increase in the absolute magnitude of the power spectrum density of the high frequency component (0.15-0. 40 Hz) of HRV and the total bandwidth (0.04-0.4 Hz) of mean BPV. These responses were related to a shift in the weighted average of the respiration frequency on the respiration spectrum, from a median value of 0.27 Hz (range, 0.17-0.29 Hz) during baseline to 0.31 Hz (0. 26-0.31 Hz) following caloric stimulation. This change was not observed in the labyrinthine-defective patient, who had weighted averages of 0.37 Hz and 0.34 Hz, respectively. No significant changes in the normalised units of the low frequency component (0. 04-0.15 Hz) or the high frequency component (0.04-0.4 Hz) of HRV and BPV were observed. During the paced breathing condition, no consistent effect on HRV or BPV was evident. For both breathing conditions, the proportions of HRV and BPV power linearly independent from respiration did not show any caloric-induced change. This study shows that caloric vestibular stimulation produces changes in HRV and BPV by modifying the respiratory pattern. PMID- 11033205 TI - Transient cardio-respiratory responses to visually induced tilt illusions. AB - Although the orthostatic cardio-respiratory response is primarily mediated by the baroreflex, studies have shown that vestibular cues also contribute in both humans and animals. We have demonstrated a visually mediated response to illusory tilt in some human subjects. Blood pressure, heart and respiration rate, and lung volume were monitored in 16 supine human subjects during two types of visual stimulation, and compared with responses to real passive whole body tilt from supine to head 80 degrees upright. Visual tilt stimuli consisted of either a static scene from an overhead mirror or constant velocity scene motion along different body axes generated by an ultra-wide dome projection system. Visual vertical cues were initially aligned with the longitudinal body axis. Subjective tilt and self-motion were reported verbally. Although significant changes in cardio-respiratory parameters to illusory tilts could not be demonstrated for the entire group, several subjects showed significant transient decreases in mean blood pressure resembling their initial response to passive head-up tilt. Changes in pulse pressure and a slight elevation in heart rate were noted. These transient responses are consistent with the hypothesis that visual-vestibular input contributes to the initial cardiovascular adjustment to a change in posture in humans. On average the static scene elicited perceived tilt without rotation. Dome scene pitch and yaw elicited perceived tilt and rotation, and dome roll motion elicited perceived rotation without tilt. A significant correlation between the magnitude of physiological and subjective reports could not be demonstrated. PMID- 11033206 TI - Autonomic nervous system activity associated with postural disturbances in patients with perilymphatic fistula: sympathetic or vagal origin? AB - The study focused on patients suffering from perilymphatic fistula (PLF), whether they had undergone surgery or not. Vestibular disturbances can be harmless but are associated with varying symptoms, demonstrating disorders within the autonomic nervous system (ANS). The aim was to test whether the orthosympathetic is involved as the vagal part is often suspected of eliciting a feeling of sickness. Non-invasive and uninterrupted recording of ANS activity represents an objectivation technique to evidence such disturbances. Electrodermal activity, thermovascular variables, instantaneous cardiac rate and blood pressure were recorded. Discomfort was triggered experimentally by applying various stimulations successively to the intact ear then to the PLF (or operated) side. Twelve subjects took part in the experiment. Two types of ANS activity were distinguished: (1) phasic responses during stimulation and (2) tonic evolution thereafter. Results show strong activation in orthosympathetic variables when the PLF side was stimulated. No further significant difference between the two sides was to be observed following surgery. After stimulation, a sudden increase in skin resistance was observed, associated with slight bradycardia. No vagal signs having been evidenced, actual nausea may result from brief inhibition of sympathetic activation resulting, in turn from primary over-activation of this system. PMID- 11033207 TI - Neck afferent involvement in cardiovascular control during movement. AB - It is well established that labyrinth and neck afferent information contributes to the regulation of somatomotor function during movement and changes in posture. There is also convincing evidence that the vestibular system participates in the modulation of sympathetic outflow and cardiovascular function during changes in posture, presumably to prevent orthostatic hypotension. However, the labyrinth organs do not provide any signals concerning body movements with respect to the head. In contrast, the neck receptors, particularly muscle spindles, are well located and suited to provide information about changes in body position with respect to the head and vestibular signals. Studies in the cat suggest that neck afferent information may modulate the vestibulosympathetic reflex responses to head-neck movements. There is some evidence in the cat to suggest involvement of low threshold mechanoreceptors. However, human studies do not indicate that low threshold mechanoreceptors in the neck modulate cardiovascular responses. The human studies are consistent with the studies in the cat in that they demonstrate the importance of otolith activation in mediating cardiovascular and sympathetic responses to changes in posture. This paper briefly reviews the current experimental evidence concerning the involvement of neck afferent information in the modulation of cardiovascular control during movement and changes in posture. PMID- 11033208 TI - Spinal modulation of the muscle pressor reflex by nitric oxide and acetylcholine. AB - Static contraction of skeletal muscle activates the sympathetic nervous system, which in turn increases cardiovascular function. These changes are mediated, in part, by a reflex arising from the contracting muscle. This reflex is termed the exercise pressor reflex or, more simply, the muscle pressor reflex (MPR). Over the past few years, studies have been performed investigating the sensory processing that occurs in the dorsal horn of the spinal cord as it pertains to the MPR. Several putative neurotransmitters and receptors have been implicated in mediating the MPR at the level of the dorsal horn. In addition, several receptor systems have been shown to modulate the MPR at the dorsal horn. We have recently performed studies investigating the potential modulatory role of dorsal horn nitric oxide (NO) and acetylcholine (ACH) on the MPR. Along these lines, our experiments suggest that NO enhances the excitability of dorsal horn cells receiving input from muscle afferent neurons, while ACH decreases the MPR when its concentration in the dorsal horn is elevated. The purpose of this manuscript is to review recently published findings from our laboratory and apply this information in an effort to better understand the integration of sensory input that occurs in the dorsal horn as it pertains to cardiovascular regulation. This review is also designed to stimulate questions as to how these two neurochemicals exert their actions and whether or not they represent or can represent important physiological mechanisms involved in regulating the dorsal horn integration of the MPR. PMID- 11033209 TI - Somatosympathetic reflex in a working heart-brainstem preparation of the rat. AB - The purpose of the present study was to examine the cardiorespiratory responses (CR) evoked by a somatosympathetic reflex (SSR) in the working heart-brainstem preparation (WHBP). Sprague-Dawley rats (75-100 g) were anesthetized with halothane, bisected sub-diaphramatically and decerebrated pre-collicularly (n = 15). The preparation was transferred to a recording chamber and perfused via the thoracic aorta with Ringer's solution containing an oncotic agent (Ficoll, 1.25%). SSR was activated by electrical stimulation (5 s) of the brachial nerve (0.5-40 Hz, 1-20 V, 0.1 ms) or the forelimb (0.5-40 Hz, 5-60 V, 2 ms). Stimulation at 40 Hz significantly increased heart rate (HR, 366 +/- 10 to 374 +/ 9 beats/min), systemic perfusion pressure (PP, 83 +/- 5 to 89 +/- 6 mmHg) and phrenic nerve discharge (PND, 0.4 +/- 0.1 to 1.4 +/- 0.3 Hz). Ganglionic blockade with hexamethonium (300 microM) eliminated the tachycardia and pressor response but did not alter the tachypnea to forelimb stimulation (n = 3). Transection of the brachial nerve plexus abolished the increase in PP and PND (n = 4). This indicates that a neural reflex mediated these responses. Spinal transection (C1 C2) completely abolished all responses indicating that they were mediated via a supraspinal pathway (n = 2). Based upon these findings, we conclude that activation of somatosensory afferent fibers in the WHBP evokes a programmed pattern of autonomic responses altering the activity-state of both the cardiovascular and respiratory systems. The WHBP provides a unique opportunity to investigate the medullary circuits and neuronal mechanisms that may be involved in coupling cardiorespiratory and somatomotor activity during locomotion/exercise. PMID- 11033210 TI - Cardiovascular responses in anticipation of changes in posture and locomotion. AB - Measurements were made in 29 adult baboons that were housed in social groups, allowing the occurrence of the full range of species-specific behavioral interactions. The cardiovascular variables measured included blood pressure, heart rate, renal blood flow, lower limb blood flow, and occasionally mesenteric blood flow. The data were telemetered from backpacks worn by the animals and were recorded in analogue form on a polygraph, digitally on a computer and were also recorded on the audio channels of videotape being made of the behavior and social interactions of the baboons. The video and the computer recordings were synchronized by a timing system that made it possible to relate the cardiovascular responses to the behavioral responses. A numerically based behavioral code was developed that allowed the categorization of the totality of the behavior, including postural and locomotor changes. Comparisons between baseline cardiovascular values and those occurring 1 s before the initiation of a movement or posture change gave no evidence of anticipatory cardiovascular responses unless the movement was associated with behavior that included emotional content. Hypothalamic perifornical lesions reduced or eliminated these anticipatory changes. PMID- 11033211 TI - Hypothalamus, hypertension, and exercise. AB - The hypothalamus is a well-known autonomic regulatory region of the brain involved in integrating several behaviors as well as cardiorespiratory activity. Our laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses associated with exercise. In addition, other findings from this laboratory and others have implicated alterations in this same brain region in spontaneously hypertensive rats as contributing factors of the elevated levels of arterial pressure in hypertension. Several studies have revealed a gamma-amino-butyric acid (GABAergic) deficiency in the caudal hypothalamus of spontaneously hypertensive rats that contributes to the tonic disinhibition and overactivity of this pressor region. Because chronic exercise is able to increase cardiovascular health in the hypertensive rat, we hypothesized that exercise induced caudal hypothalamic plasticity partially underlies the beneficial effects of physical activity. In this review we discuss initial findings from this lab that support this hypothesis. Our experiments demonstrate that chronic exercise alters gene expression and neuronal activity in the caudal hypothalamus of the spontaneously hypertensive rat. These findings describe a potential mechanism by which chronic exercise lowers blood pressure in the hypertensive individual. PMID- 11033212 TI - Cortical control of somato-cardiovascular integration: neuroanatomical studies. AB - This paper will discuss experiments dedicated to the exploration of pathways linking the sensorimotor cortex (SMC) and the main bulbar nuclei involved in cardiovascular control: the nucleus tractus solitarius (NTS), the dorsal nucleus of the vagus (DMV) and the rostral ventrolateral medulla (RVLM). Results obtained through neurofunctional and neuroanatomical methods are presented in order to bring new answers to relevant points concerning somato-cardiovascular integration: firstly to show the ability of the SMC to influence neurons in bulbar cardiovascular nuclei, and secondly to identify pathways that transmit such influences. The neurofunctional approach, based on the identification of Fos like immunoreactive neurons, indicated that the SMC has functional connections with cardiovascular bulbar nuclei. The neuroanatomical approach, which employed retrograde and anterograde axonal tracing methods, provided evidence of direct projections from the SMC to NTS/DMV and RVLM. Furthermore, experiments showed clearly that corticospinal neurons sent collaterals to bulbar cardiovascular nuclei, especially to the RVLM. Direct cortical projections to the NTS/DMV and the RVLM provide the anatomical basis for cortical influences on the baroreceptor reflex and sympathetic vasomotor mechanisms for blood pressure control, and support the hypothesis of cortical commands coupling somatic and cardiovascular outputs for action. PMID- 11033213 TI - Central circuits mediating patterned autonomic activity during active vs. passive emotional coping. AB - Animals, including humans, react with distinct emotional coping strategies to different sets of environmental demands. These strategies include the capacity to affect appropriate responses to "escapable" or "inescapable" stressors. Active emotional coping strategies--fight or flight--are particularly adaptive if the stress is escapable. On the other hand, passive emotional coping strategies quiescence, immobility, decreased responsiveness to the environment-are useful when the stress is inescapable. Passive strategies contribute also to facilitating recovery and healing once the stressful event is over. Active vs. passive emotional coping strategies are characterised further by distinct patterns of autonomic change. Active strategies are associated with sympathoexcitation (hypertension, tachycardia), whereas passive strategies are associated with sympathoinhibitory patterns (hypotension, bradycardia). Distinct neural substrates mediating active vs. passive emotional coping have been identified within the longitudinal neuronal columns of the midbrain periaqueductal gray region (PAG). The PAG offers then a potentially useful point of entry for delineating neural circuits mediating the different forms of emotional coping and their associated patterns of autonomic activity. As one example, recent studies of the connections of orbital and medial prefrontal cortical (PFC) fields with specific PAG longitudinal neuronal columns are reviewed. Findings of discrete orbital and medial PFC projections to different PAG columns, and related PFC and PAG columnar connections with specific subregions of the hypothalamus, suggest that distinct but parallel circuits mediate the behavioural strategies and patterns of autonomic activity characteristic of emotional "engagement with" or "disengagement from" the external environment. PMID- 11033214 TI - Changes in autonomic and EEG patterns induced by hypnotic imagination of aversive stimuli in man. AB - Autonomic and electroencephalographic (EEG) responses to aversive stimuli presented by means of hypnotic suggestion have been studied in man.Healthy volunteers with simple phobia were screened for susceptibility to hypnosis. The experimental paradigm included periods of rest during which the hypnotized subjects were asked to produce an emotionally neutral mental image and periods of emotional activation in which they were asked to image a phobic object. Heart rate (HR), respiratory frequency (RF) and EEG were processed to obtain the HR related indexes of sympatho-vagal balance and the EEG spectral components. The results showed a significant increase in HR and RF with a shift of the sympatho vagal indexes towards a sympathetic predominance during the hypnotic emotional activation. EEG activity showed a significant increase in the gamma band with a left fronto-central prevalence. There was also a less pronounced increase in the beta band. In conclusion, by means of hypnosis, autonomic and behavioral responses to fear-like stimuli can be induced in man in a reproducible and controlled manner. Such a paradigm could be applied in human neuroimaging studies to identify central nervous structures that modulate stress and fear-related reactions. PMID- 11033215 TI - Cerebral vasoconstriction precedes orthostatic intolerance after parabolic flight. AB - The effects of brief but repeated bouts of micro- and hypergravity on cerebrovascular responses to head-up tilt (HUT) were examined in 13 individuals after (compared to before) parabolic flight. Middle cerebral artery mean flow velocity (MCA MFV; transcranial Doppler ultrasound), eye level blood pressure (BP) and end tidal CO(2) (P(ET)CO(2)) were measured while supine and during 80 degrees HUT for 30 min or until presyncope. In the postflight tests subjects were classified as being orthostatically tolerant (OT) (n = 7) or intolerant (OI) (n = 6). BP was diminished with HUT in the OT group in both tests (p < 0.05) whereas postflight BP was not different from supine in the OI group. Postflight compared to preflight, the reduction in P(ET)CO(2) with HUT (p < 0.05) increased in both groups, although significantly so only in the OI group (p < 0.05). The OI group also had a significant decrease in supine MCA MFV postflight (p < 0.05) that was unaccompanied by a change in supine P(ET)CO(2). The decrease in MCA MFV that occurred during HUT in both groups preflight (p < 0.05) was accentuated only in the OI group postflight, particularly during the final 30 s of HUT (p < 0.05). However, this accentuated decrease in MCA MFV was not correlated to the greater decrease in P(ET)CO(2) during the same period (R = 0.20, p = 0.42). Although cerebral vascular resistance (CVR) also increased in the OI group during the last 30 s of HUT postflight (p < 0.05), the dynamic autoregulatory gain was not simultaneously changed. Therefore, we conclude that in the OI individuals, parabolic flight was associated with cerebral hypoperfusion following a paradoxical augmentation of CVR by a mechanism that was not related to changes in autoregulation nor strictly to changes in P(ET)CO(2). PMID- 11033216 TI - A role for adenosine in modulating cardio-respiratory responses: a mini-review. AB - Evidence is provided that purine nucleotides play a major role in modulating cardio-respiratory control through actions at specific sites in the central nervous system (CNS). The accumulation of adenosine in the extracellular space of the nucleus tractus solitarius (NTS) on hypothalamic defence area stimulation is described, and is shown to result from the extracellular breakdown of neurally released adenosine triphosphate (ATP). The actions of adenosine in modulating synaptic actions in the NTS are discussed in relation to the potential presynaptic action of adenosine in modulating transmitter release. Actions of adenosine and ATP at other sites in the medulla concerned with cardio-respiratory control are discussed. The inter-relationships between ATP and adenosine are mentioned. PMID- 11033217 TI - Visualization of sleep influences on cerebellar and brainstem cardiac and respiratory control mechanisms. AB - Cerebellar and vestibular structures exert substantial influences on breathing and cardiovascular activity, particularly under conditions of extreme challenges. Influences from these structures, as well as from the ventral medullary surface, are greatly modified during sleep states. Vestibular lesions abolish the pronounced phasic autonomic variation found in the rapid eye movement sleep state, and spontaneous ventral medullary surface activity, as assessed by optical procedures, is greatly diminished in that state. Neural responses from the ventral medullary surface to hypotensive challenges are enhanced and appear "undampened" during the rapid eye movement sleep state. Functional magnetic resonance imaging reveals activation to blood pressure challenges in widespread brain areas of humans, and especially in cerebellar sites, such as the fastigial nucleus. A subset of victims of sudden infant death syndrome, a sleep-related disorder, appear to succumb from cardiovascular failure of a shock-like nature, and often show neurotransmitter receptor deficiencies in the ventral medullary surface, caudal midline raphe hypotensive regions, and the inferior olive, a major afferent relay to the cerebellum. Afferent and efferent vestibular/cerebellar structures, or sites within the cerebellum may mediate failure mechanisms in sudden infant death syndrome and a number of other sleep disordered breathing and cardiovascular syndromes. PMID- 11033228 TI - The effect of gender, sexual maturation and xenobiotic treatment on the formation of hydroxymethyl metabolites from 7, 12-dimethylbenz[a]anthracene in rat liver microsomes. AB - The effects of age, gender, strain, phenobarbital (PB) treatment and pituitary influence on the regioselective metabolism of 7, 12-dimethylbenz[a]anthracene to hydroxmethyl metabolites were investigated. Studies used hepatic microsomal membranes from immature and mature Long Evans (LE) rats and adult Hooded Lister (HL) animals. Hydroxymethyl metabolites were resolved by both normal and reverse phase HPLC with on-line diode array detection. The CYP isoform(s) responsible for oxidation at the 12 methyl position exhibited no gender or developmental regulation and the rate of formation was not altered following hypophysectomy. PB treatment of adult rats caused a significant increase in the rate of formation of both male and female animals (29 and 41-fold, respectively) suggesting a major contribution from a PB-inducible isoform, such as CYP2B. The rate of formation of 7OHMe12MBA exhibited no gender dependency in immature animals but was 2-fold greater than that observed for 12OHMe7MBA suggesting that steric hindrance resulting from the adjacent 1,2 benzyl ring favours substrate oxidation at the 7 methyl position. Male predominant formation of 7OHMe12MBA was apparent following sexual maturation of the LE rats and was significantly reduced upon hypophysectomy suggesting the involvement of a male-specific GH dependent isoform e.g. CYP2C11. PMID- 11033229 TI - Interactions between benzo[a]pyrene and UVA light affecting ATP levels, cytoskeletal organization, and resistance to trypsinization. AB - Polycyclic aromatic hydrocarbons affect cells in many ways, including covalent modifications of DNA, participation in redox cycling, and alterations in cellular signaling pathways. Similarly, exposure to ultraviolet (UV) light may modify DNA, generate reactive oxygen species, and alter signaling. Because environmental conditions may interact to affect cellular functions, we investigated the combined effects of benzo[a]pyrene (BaP) and UV light in a cell line in which BaP induced alterations in Ca(2+) homeostasis have previously been shown. Exposure of MCF-10A cells to BaP (18 h) followed by a brief (5 min) exposure to UVA resulted in resistance to trypsinization of cells grown on type I collagen (Vitrogen). This effect was not seen following treatment with BaP or UVA alone nor with benzo(e)pyrene (BeP)+UVA. BaP+UVA light also caused actin filaments to reorganize from typical stress fibers to substrate-associated aggregates of actin and caused depletion of cellular adenosine triphosphate (ATP). The effects of BaP+UVA on adhesion and actin aggregate formation were partially prevented by treatment with reduced glutathione. Depletion of cellular ATP affected resistance to trypsinization and actin organization in a similar manner. Thus, these studies suggest a redox-sensitive interaction between BaP+UVA light to deplete cellular ATP levels, resulting in resistance to trypsinization and actin filament reorganization in MCF-10A cells. PMID- 11033230 TI - Polycyclic aromatic hydrocarbon coated onto Fe(2)O(3) particles: assessment of cellular membrane damage and antioxidant system disruption in human epithelial lung cells (L132) in culture. AB - The aim of this study was to investigate the oxidative effects of Fe(2)O(3), benzo(a)pyrene (B(a)P) and pyrene, alone or in association (B(a)P or pyrene coated onto Fe(2)O(3) particles), in normal human embryonic lung epithelial cells (L132) in culture. We evaluated: (i) membrane integrity, through fatty acid release (stearic acid, oleic acid, linoleic and linolenic acids, homolinolenic acid, arachidonic acid) and malondialdehyde (MDA) production; and (ii) antioxidant status, through enzymatic and non-enzymatic antioxidant defenses (superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione status, beta-carotene). Fe(2)O(3) did not induce any change in L132 cells. In pyrene-treated cells, SOD induction (P<0. 05), glutathione oxidation (P<0.05) and beta-carotene consumption (P<0.001) may counteract free radicals (FR)-induced damage. However, in B(a)P-incubated cells, SOD inactivation (P<0.05), GR increases (P<0.05), glutathione oxidation (P<0.05) and beta-carotene decreases (P<0.001) showed high disruption of antioxidants, thereby allowing FR induced damage (i.e. arachidonic acid release, P<0.01; MDA production, P<0.01). Our main finding was that both associations caused higher FR-induced damage (i.e. MDA production, P<0.001; SOD inactivation, P<0.01) than either chemical alone. Several mechanisms could account for this result: enhanced uptake of Fe(2)O(3) particles and/or greater availability of polycyclic aromatic hydrocarbons (PAHs). We hypothesized also that Fe(2)O(3) and polycyclic aromatic hydrocarbons are more deleterious by virtue of their associations being able to produce higher oxidative effects than either chemical alone. PMID- 11033231 TI - Evidence that kynurenine pathway metabolites mediate hyperbaric oxygen-induced convulsions. AB - Metabolism of tryptophan (TRP) through the kynurenine (KYN) pathway in brain, liver, and kidney produces intermediates including the neuroactive agonist quinolinic acid (QA) and the antagonists kynurenic acid (KA) and anthranilic acid (AA) for N-methyl D-aspartate (NMDA) receptors in the central nervous system. We hypothesized that elevated concentrations of QA, KA, or AA can moderate the convulsions that are observed during exposure of rats to hyperbaric oxygen (HBO). We found that i.p. administration of TRP or KYN (both of which cross the blood brain barrier) had no effect on HBO-induced seizures. However, AA (administered i.p.) or gavage administration of the KYN pathway blocking drug Ro 61-8048, both of which enter the brain from the circulatory system, affect the time to first convulsion and/or coma during HBO in a manner consistent with a modulatory role for seizure activity. PMID- 11033232 TI - Effect of aluminum on delta-aminolevulinic acid dehydratase from mouse blood. AB - The objective of this study was to investigate aluminum deposition in whole blood and plasma of mice and the activity of blood delta-aminolevulinic acid dehydratase (ALA-D) after in vitro and in vivo exposure to this element. In vitro experiments showed activation and inhibition of the enzyme activity when 0.01-5.0 mM of aluminum sulphate were used (IC(50): 1.31 mM). Treatment with citrate and aluminum plus citrate increased ALA-D activity in vivo and the increase in enzyme activity was parallel to the increase in aluminum content in blood and plasma. These results show that aluminum has a distinct effect on ALA-D activity: first, at relatively lower concentrations it activated, and at high concentration it inhibited, blood ALA-D in vitro; second, it activated the enzyme when administered to drinking water. One important toxicological finding of the present report is that the apparent irrelevant addition of citrate to the drinking water significantly increased the level of aluminum in blood and plasma. Thus, in order to predict more accurately the extent of human exposure to aluminum it would be advantageous to consider the level of citrate ingestion and not exclusively the aluminum level in water or food. PMID- 11033233 TI - N-acetyl L-cysteine attenuates oxidant-mediated toxicity induced by chrysotile fibers. AB - Chrysotile, an important commercial variety of asbestos, is known to cause oxidative stress by enhancing production of hydrogen peroxide (H(2)O(2)) and thiobarbituric acid reactive substances (TBARS), depleting glutathione (GSH) and altering levels of GSH redox system enzymes. N-acetyl L-cysteine (NAC), a compound that increases GSH levels, protects cells against chrysotile toxicity. In the present study, rats were exposed intratracheally to a single dose (5 mg/rat) of chrysotile. This was followed by a daily dose of NAC 50 mg/kg. b. wt., i.p. At 1, 4, 8 and 16 days post chrysotile exposure lung lavage fluid was collected to determine H(2)O(2) generation, TBARS production, GSH level and its redox system enzymes activities. A significant decrease in H(2)O(2) and TBARS, an increase in GSH content and its redox system enzymes was observed in chrysotile+NAC animals in comparison to chrysotile-exposed animals. In this preliminary study it appears that NAC may be protecting cells against oxidative damage. This protection may be due to its ability to maintain intracellular GSH/oxidative scavenging capability. PMID- 11033234 TI - Effects of arsenite on central monoamines and plasmatic levels of adrenocorticotropic hormone (ACTH) in mice. AB - We studied the effects of chronic arsenic exposure on brain monoamines and plasma levels of adrenocorticotropic hormone (ACTH) of mice. After weaning, mice received arsenic (0, 20, 40, 60 or 100 ppm) in drinking water over a period of 9 weeks. Monoamine content was quantified in different brain regions, arsenic was quantified in brain tissue and ACTH levels in plasma. Brain arsenic concentrations up to 200 ng/g showed a significant correlation with exposure levels and produced slight modifications in regional monoamine levels. ACTH plasma levels were significantly associated with norepinephrine (NE) concentrations in the medulla and pons, but not with hypothalamic NE levels. ACTH levels were significantly higher in the group exposed to 20 ppm. Dopamine showed significant dose-related decreases in the hypothalamus. These results show that chronic sodium arsenite exposure produces changes in central monoamines, which are not associated on a dose-dependent basis with major alterations in plasma ACTH. PMID- 11033236 TI - Effects of 2-ethylhexanoic acid on the production of reactive oxygen species in human polymorphonuclear leukocytes in vitro. AB - 2-Ethylhexanoic acid (2-EHA), is an industrial chemical and a toxic biotransformation product of the plasticizer di(2-ethylhexyl)phthalate. Its immunological effects are unknown. 2-EHA resembles structurally C18 fatty acids, which are known activators of respiratory burst in human polymorphonuclear leukocytes (PMNL). Therefore, we exposed PMNL to 2-EHA in vitro and measured the production of reactive oxygen species (ROS) and explored the associated cellular mechanisms. 2-EHA (10-2000 microM) inhibited dose-dependently formyl-methionyl leucyl-phenylalanine (FMLP)-induced respiratory burst in PMNL. Moreover, 2-EHA decreased oxidative burst evoked by the protein kinase C (PKC) activators, phorbol myristate acetate (PMA) and dioctanoyl-s,n-glycerol (DIC(8)). 2-EHA affected neither the levels of free intracellular calcium nor inhibited PKC. The results indicate that 2-EHA inhibits activation of PMNL to produce ROS, i.e. has an immunosuppressive effect in vitro. The site of action in the PKC is after activation of this enzyme. PMID- 11033235 TI - Induction of cytochrome P450 1A1 in human hepatoma HepG2 cells by 6 nitrochrysene. AB - The present study has determined the effects of 6-nitrochrysene (6-NC) on human cytochrome P450-dependent monooxygenases in human hepatoma HepG2 cells. Treatment of HepG2 cells with 6-NC increased the activities of microsomal benzo[a]pyrene hydroxylase, 7-ethoxycoumarin and 7-ethoxyresorufin O-deethylases, cytosolic glutathione S-transferase and N-acetyltransferase, and S9 metabolic activation of 6-NC in the Ames mutagenicity test. Immunoblot and RNA blot analyses revealed that 6-NC induced CYP1A1 protein and mRNA levels in the hepatoma cells. Nuclear transcription assay demonstrated that 6-NC increased the transcription rate of CYP1A1 gene in HepG2 cells. Treatment of human lung carcinoma NCI-H322 cells with 6-NC increased benzo[a]pyrene hydroxylase activity and CYP1A1 protein and mRNA levels. These results demonstrate that 6-NC is an inducer of human CYP1A1 and the induction occurs at a transcriptional level in HepG2 cells. The ability of 6-NC to induce liver and lung CYP1A1 may be an important factor to consider in assessing 6-NC metabolism and toxicity in humans. PMID- 11033237 TI - Gossypol-induced DNA breaks in rat lymphocytes are secondary to cytotoxicity. AB - Gossypol, a male antifertility and potential anticancer agent, was found to induce DNA strand breaks in rat lymphocytes. DNA breaks were measured with the single-cell gel electrophoresis (SCGE) or 'comet' assay. A significant increase in DNA breaks was observed after 1 h incubation at concentrations of 2 microg/ml or greater. The inclusion of 10% fetal bovine serum in the media reduced the toxicity of gossypol, and DNA breaks were only observed at a concentration of 80 microg/ml. However, the increase in DNA strand breaks, for incubations with and without serum, only occurred when cell viability was reduced to less than 70%. Examination of cell morphology and DNA fragmentation at incubations up to 5 h yielded no evidence that DNA strand breaks were occurring due to apoptosis. We conclude that gossypol is not primarily genotoxic in this cell type, and that the DNA breaks observed arose secondary to cytotoxicity. PMID- 11033238 TI - Kupffer cells mediate increased anoxic hepatocellular killing from hyperosmolarity by an oxygen- and prostaglandin-independent mechanism. AB - The purpose of this study was to evaluate the roles of Kupffer cells, prostaglandin biosynthesis, and glycolytic metabolism in accelerated anoxic cell killing by hyperosmolar stress. Isolated rat livers were perfused with anoxic normosmolar Krebs-Heinseleit bicarbonate buffer (KHB) or anoxic hyperosmolar KHB (+40 mM NaCl). Hyperosmolar KHB accelerated LDH release during anoxia in livers from both fed and fasted rats by as much as 3.7-fold. GdCl(3) pretreatment to inactivate Kupffer cells substantially delayed anoxic LDH release during normosmolar perfusions and blocked entirely the hyperosmolarity-induced acceleration of LDH release. Cyclooxygenase inhibition with indomethacin failed to alter LDH release during anoxia in hyperosmolar KHB. Neither GdCl(3) nor hyperosmolarity changed glycolytic flux during hypoxia, and hyperosmolarity did not change basal oxygen uptake. We conclude that accelerated cell killing in hyperosmolar buffer is a Kupffer cell-dependent event that is independent of oxygen-requiring prostaglandin synthesis, changes of glycolytic flux, and activation of cellular ATP demand. Another as yet unidentified Kupffer cell product appears to mediate the effect of hyperosmolarity of anoxic hepatocellular injury. PMID- 11033240 TI - Pharmacokinetics and metabolism of ethylenediamine in the swiss webster mouse following oral or intravenous dosing. AB - Male Swiss Webster mice were given an intravenous dose of 50 mg/kg, or an oral gavage dose of 5, 50 or 500 mg/kg [1, 2-(14)C]-ethylenediamine dihydrochloride, and its fate was followed for 48 h. Ethylenediamine (EDA) was readily absorbed from the gut (bioavailability, 87% measured at 50 mg/kg). Absorption was rapid as the EDA concentration in plasma reached a maximum at about 1 h after dosing. 14C EDA-derived radioactivity was distributed throughout the body, with the liver and kidney attaining the highest concentration among the major organs. Urine was the major route of excretion, accounting for over half of the dose. About 4-13 and 8% of the dose was eliminated in the feces and as expired CO(2), respectively. Excretion was quite rapid, with over 70% of the applied dose eliminated within 24 h. The principal metabolite in the urine was N-acetylethylenediamine. There was some indication that the metabolism of EDA in the mouse might be saturated at 500 mg/kg, as the percentage of N-acetylethylenediamine excreted in the urine decreased markedly, with a concomitant shift to a higher proportion of unchanged EDA, when compared with the lower dosages. PMID- 11033239 TI - Prenatal exposure model simulating CO inhalation in human cigarette smokers: sphingomyelin alterations in the rat sciatic nerve. AB - Prenatal exposure to low concentrations of carbon monoxide (CO, 150 ppm) causes long-term alterations in sphingomyelin (SM) homeostasis in peripheral nervous system, but not brain of male rat offspring. In particular, unlike sphinganine (intermediate of complex sphingolipid biosynthesis de novo), the concentrations of sphingosine (intermediate of complex sphingolipid turnover) were increased by 2.35-fold in the sciatic nerve of CO-exposed offspring with respect to controls (P<0.05, overall one-way ANOVA). These subtle alterations were not accompanied by changes in motor activity (F=0.25, df=1/10, n.s., overall one-way-ANOVA). The results suggest that the SM homeostasis in the sciatic nerve is particularly susceptible to prenatal CO exposure resulting in maternal carboxyhaemoglobin (HbCO) levels equivalent to those found in human cigarette smokers. PMID- 11033241 TI - Styrene inhibits monoamine oxidase A, but not monoamine oxidase B in monkey brain mitochondria. AB - The effects of styrene on mitochondrial monoamine oxidase (MAO) activity in rat and monkey brains were compared in vitro. After preincubation at 25 degrees C for 20 min with 1 mM styrene monomer MAO-A activity in monkey brain was inhibited potently using 5-HT (for MAO-A substrate), but MAO-B activity in monkey brain and platelets were slightly inhibited using beta-PEA (for MAO-B substrate). Styrene monomer also competitively inhibited MAO-A activity in a dose-dependent manner. MAO-A in monkey brain was inhibited by styrene in ascending order of potency: styrene trimer>styrene dimer>styrene monomer. In contrast styrene monomer slightly inhibited both MAO-A and MAO-B activities in rat brain mitochondria. In the present study styrene monomer potently inhibits MAO-A activity, but not MAO-B activity, in monkey brain mitochondria in vitro. These results indicate the inhibiting action of styrene differs depending on animal species and MAO isoforms. PMID- 11033242 TI - Evolution of hearing in vertebrates: the inner ears and processing. AB - This paper considers aspects of the evolution of the vertebrate auditory system from an 'ichthyocentric' perspective. It is argued that all vertebrate auditory systems are required to do certain basic tasks including acoustic feature discrimination, sound source localization, frequency analysis, and auditory scene analysis, among others. These sorts of capabilities arose very early in the evolution of the vertebrates and have been modified by selection in different species. In some cases the same structures have been involved in detection and analysis throughout the vertebrates, while in other cases the mechanism by which the same type of analysis takes place may have changed. PMID- 11033243 TI - Effects on cochlear responses of activation of descending pathways from the inferior colliculus. AB - The inferior colliculus (IC) has been shown anatomically to make direct descending connections with medial olivocochlear (MOC) neurones in the auditory brainstem. The MOC neurones project to the outer hair cells in the cochlea and inhibit cochlear neural output. This study investigated the effect of IC stimulation on cochlear output in both guinea pigs and rats in order to determine the functional significance of the IC-to-olivocochlear system projection. Stimulation of the central nucleus and the external cortex of the IC in paralysed guinea pigs, both contra- and ipsilaterally to the test cochlea, resulted in a small increase of the cochlear microphonic amplitude and a small decrease of the compound action potential (CAP) amplitude, the latter equivalent to a 3-6 dB change in acoustic input. Effects on the CAP were maximal in the frequency range 6-10 kHz. These effects were consistent with partial activation of the MOC system. In unparalysed rats, stimulation of the inferior colliculus evoked a large, prolonged suppression ranging from 5-12 dB in the amplitude of distortion product otoacoustic emissions (2f(1)-f(2); DPOAE), as reported previously by Scates et al. (1999). However, this suppression was decreased to only 0-3 dB when the animals were paralysed, suggesting that the larger suppression in the unparalysed state was the consequence of either a general masking effect caused by animal movement, or activation of middle ear muscles by the inferior colliculus stimulation. The results indicate a small but significant excitatory effect of the inferior colliculus on the medial olivocochlear system under conditions of anaesthesia and paralysis. PMID- 11033244 TI - An auditory negative after-image as a human model of tinnitus. AB - The Zwicker tone (ZT) is an auditory after-image, i.e. a tonal sensation that occurs following the presentation of notched noise. In the present study, the hypothesis that neural lateral inhibition is involved in the generation of this auditory illusion was investigated in humans through differences in perceptual detection thresholds measured following broadband noise, notched noise, and low pass noise stimulation. The detection thresholds were measured using probe tones at several frequencies, within as well as outside the suppressed frequency range of the notched noise, and below as well as above the corner frequency of the low pass noise. Thresholds measured after broadband noise using a sequence of four 130-ms probe tones (with a 130-ms inter-burst interval) proved to be significantly smaller that those measured using the same probe tones after notched noise at frequencies falling within the notch, but larger for frequencies on the outer edges of the noise. Thresholds measured following low-pass noise using the same sequence of probe tones were found to be smaller at frequencies slightly above the corner, but larger at lower, neighboring frequencies. This pattern of results is consistent with the hypothesis that the changes in auditory sensitivity induced by stimuli containing sharp spectral contrasts reflect lateral inhibition processes in the auditory system. The potential implications of these findings for the understanding of the mechanisms underlying the generation of auditory illusions like the ZT or tinnitus are discussed. PMID- 11033245 TI - Developmental changes in physiological properties in the rat's dorsal nucleus of the lateral lemniscus. AB - To better understand the development of the dorsal nucleus of the lateral lemniscus (DNLL), intrinsic membrane properties and synaptic responses of DNLL neurons in brain slice preparations were examined. Intracellular recordings were taken from DNLL neurons of rat pups at postnatal days 4-8 (early group), 10-12 (intermediate group) and 16-18 (late group). In response to positive current injection, neurons in the early group displayed firing with lower frequency and a longer action potential duration in comparison to the intermediate and late groups. The action potential amplitude of DNLL neurons increased during development. Postsynaptic potentials (PSPs), with excitatory predominance, were elicited by electrical stimulation of the lateral lemniscus and commissure of Probst throughout the three age groups. Neurons showed a longer latency and rise time of the PSPs in the early group in comparison with those in the intermediate and late groups. These results suggest that the early DNLL neurons display physiological characteristics associated with immature neurons, while the other two groups tend to illustrate mature-like neuronal properties. Furthermore, it seems that the neurons at day 10-12 are in a transitional period of development, which coincides with the onset of hearing. PMID- 11033246 TI - Ionic and potential changes of the endolymphatic sac induced by endolymph volume changes. AB - The endolymphatic sac (ES) is believed to be the locus for endolymph volume regulation in the inner ear. It has recently been shown that induced endolymph volume changes in the cochlea result in anatomical changes in the ES, suggesting that function of the sac varies according to endolymph volume status. In the present study we have recorded luminal concentrations of K(+) and Na(+) from the ES and the endolymphatic sac potential (ESP) during cochlear endolymph volume changes. ES recordings were made by an extradural approach, thereby preserving normal cerebrospinal fluid resting pressure. Cochlear endolymph volume changes were generated by performing injections or withdrawals through a pipette inserted into endolymph by a round window approach. The pre-treatment concentrations of K(+) and Na(+) in the ES were found to be 8.4 mM (S.D. 3.3, n=8) and 128. 6 mM (S.D. 18.4, n=10) respectively, and the mean ESP was 14.4 mV (S. D. 5.2, n=18). Endolymphatic injections were found to produce a sustained increase in the K(+) content of the ES by an average of 19. 9 mM and to decrease Na(+) by 30.7 mM measured 50 min after the start of injection. The time for K(+) increase to occur was found to correlate with the injected volume, with larger injected volumes producing a more rapid increase. Endolymphatic withdrawals were found to induce a slow decline in endolymphatic K(+) by an average of 3.4 mM measured at 50 min after withdrawal, although no significant change of Na(+) was detected. Volume induced ESP changes were highly variable. Injections produced a small increase in the mean ESP and withdrawals produced a small decrease but neither change was statistically significant and some animals showed potential changes in the opposite direction. These data show that a change in cochlear endolymph volume status results in a physiologic response of the ES which is sustained for a considerable period. If the ES plays a part in the restoration of normal endolymph volume, this process appears to proceed slowly, based on the prolonged time courses of ionic changes observed. PMID- 11033247 TI - Amplification in the apical turn of the cochlea with negative feedback. AB - The apical turn of the anesthetized guinea pig cochlea was opened to examine the basilar membrane optically through the intact Reissner's membrane. Vibrations of the outer Hensen's cell and the basilar membrane (BM) adjacent to and about 130 microm below the level of the Hensen's cell were measured. Outer Hensen's cell vibration at the characteristic frequency was up to 900 times higher compared to the BM amplitude. After sacrifice BM vibration increased while Hensen's cell vibration decreased. The magnitude and sequence of change after sacrifice can best be explained by the presence of negative feedback between reticular lamina and BM. In other experiments using ototoxic drugs that damage outer hair cells, similar changes in Hensen's cell and BM vibration were observed. These results show that the apical turn behavior is different from that observed by other investigators in the basal turn. The potential benefits of the negative feedback are discussed. The presence of negative feedback would explain the linearity at the fundamental frequency observed in the apical turn of cochlea. PMID- 11033248 TI - Model calculations of time dependent responses to binaural stimuli in the dorsal nucleus of the lateral lemniscus. AB - In a previous paper (Reed and Blum, 1999), we examined the connectional hypotheses put forward by Markovitz and Pollak (1994) to explain the steady-state behavior of cells in the dorsal nucleus of the lateral lemniscus (DNLL). We found that the steady-state outputs of the four major binaural types of cells found in the DNLL (EI, EI/F, EE/I, and EE/FI) could be accounted for by known connectional patterns using only one or two cells per nucleus and quite simple hypotheses on cell behavior. In this study, we examine the time course of DNLL outputs in response to constant, ongoing, monaural or binaural sounds of various intensities. The model auditory nerve fibers ramp up linearly (usually in 2 ms) to full firing and the anteroventral cochlear nucleus cells have primary-like discharge patterns. Fixed time delays of 1 ms at each synapse are included; other time delays are employed when necessary to understand and reproduce specific features of the experimental data. We find that the connectional patterns utilized in our previous study can account for the rich variety of temporal response patterns found experimentally in the DNLL. Our main findings are: (1) all of the four major binaural types of cells can arise from modifications of the basic connectional pattern that produces EI cells; (2) both excitation and inhibition from the ipsilateral lateral superior olive (LSO) are required to understand DNLL responses; (3) pauser behavior can arise either from time delayed inhibition from a DNLL interneuron or by projection from the LSO; (4) two different mechanisms can account for the ipsilaterally evoked onset response; (5) to explain completely the temporal discharge pattern and binaural interactions of EE/FI cells, a projection from the contralateral DNLL via the commissure of Probst is necessary. PMID- 11033249 TI - Effects of deafferentation on the electrophysiology of ventral cochlear nucleus neurons. AB - When cochlear pathology impairs the afferent innervation of the ventral cochlear nucleus (VCN), electrical responses of the auditory brainstem are altered and changes in cell and synaptic morphology are observed. However, the impact of deafferentation on the electrical properties of cells in the VCN is unknown. We examined the electrical properties of single neurons in the anterior and posterior VCN following bilateral cochlear removal in young rats. In control animals, two populations of cells were distinguished: those with a linear subthreshold current-voltage relationship and repetitive firing of action potentials with regular interspike intervals (type I), and those with rectifying subthreshold current-voltage relationships and phasic firing of 1-3 action potentials (type II). Measures of action potential shape further distinguished these two groups. Two weeks following cochlear removal, both electrical response patterns were still seen. Type I cells showed a higher input resistance. Deafferented single-spiking type II cells were slightly more depolarized, had smaller action potentials, smaller afterhyperpolarizations and shorter membrane time constants, whereas multiple-spiking type II cells were apparently unaffected. These changes in the electrical properties of VCN neurons following cochlear injury may adversely affect central processing of sounds presented acoustically or electrically by prostheses. PMID- 11033250 TI - Comparison of frequency discrimination thresholds for complex and single tones in chinchillas. AB - Frequency discrimination thresholds were measured from five chinchillas for harmonic tone complexes having a fundamental frequency of 250 Hz. Stimuli consisted of the fundamental frequency and the second through 10th harmonics with individual components added in either cosine phase or random phase. In general, thresholds were independent of overall level for sound levels between 47 and 77 dB SPL, and there was no difference in thresholds observed between cosine-phase tone complexes and random-phase tone complexes. Discrimination thresholds were also obtained for a single 250-Hz tone for comparison with complex tone thresholds. Similar to data reported in human subjects, thresholds in chinchillas for tone complexes were lower than thresholds obtained using a single tone, although chinchillas required a larger frequency difference than human listeners. The results suggest that the mechanisms of frequency discrimination of complex tones are similar between chinchillas and human listeners. PMID- 11033251 TI - The neuronal response to electrical constant-amplitude pulse train stimulation: evoked compound action potential recordings. AB - The purpose of this study was to gain a greater understanding of the electrically evoked compound action potential (EAP) responses to pulse train stimulation. Analysis of EAP amplitude responses suggested that an alternating pattern varied depending upon stimulus level, interpulse interval (IPI), stimulus waveform, and stimulus polarity. Stimulus level-dependent recovery was seen in the cat and the guinea pig: higher stimulus level tended to provide faster recovery. Both polarity-dependent recovery and polarity-dependent adaptation were observed in the cat and these stimulus polarity effects were less consistent in the guinea pig. The polarity-dependent recovery effect supports the hypothesis that anodal and cathodal stimuli excite different sites along auditory nerve fibers. Amplitude differences between the response to the second pulse and the steady state response at the same IPI are significantly greater for anodal stimuli than for cathodal stimuli in all cats. These data suggest that there is a cumulative refractory effect in the auditory nerve of cats, especially in response to anodal stimuli. PMID- 11033252 TI - The neuronal response to electrical constant-amplitude pulse train stimulation: additive Gaussian noise. AB - Experimental results from humans and animals show that electrically evoked compound action potential (EAP) responses to constant-amplitude pulse train stimulation can demonstrate an alternating pattern, due to the combined effects of highly synchronized responses to electrical stimulation and refractory effects (Wilson et al., 1994). One way to improve signal representation is to reduce the level of across-fiber synchrony and hence, the level of the amplitude alternation. To accomplish this goal, we have examined EAP responses in the presence of Gaussian noise added to the pulse train stimulus. Addition of Gaussian noise at a level approximately -30 dB relative to EAP threshold to the pulse trains decreased the amount of alternation, indicating that stochastic resonance may be induced in the auditory nerve. The use of some type of conditioning stimulus such as Gaussian noise may provide a more 'normal' neural response pattern. PMID- 11033253 TI - Reduction of noise-induced hearing loss using L-NAC and salicylate in the chinchilla. AB - The effects of a combination of two antioxidant compounds were studied in a chinchilla model of noise-induced hearing loss. After obtaining baseline hearing thresholds using inferior colliculus evoked potentials, chinchillas were exposed for 6 h to octave band noise centered at 4 kHz (105 dB SPL). Post-noise thresholds were obtained 1 h after the noise exposure, and then animals received either saline or salicylate and N-L-acetylcysteine combination. Another group received antioxidant treatment 1 h prior to noise. Hearing was tested at 1, 2 and 3 weeks post-noise. Subsequently, the cochleae were harvested, and cytocochleograms were prepared. There was a 20-40 dB SPL threshold shift at 3 weeks for tested controls. Permanent threshold shifts (PTS) were significantly reduced (P<0.05) to approximately 10 dB for the pre-treatment group at week 3. The PTS for the post-treatment group at week 3 was similar to the pre-treatment group at 1 and 2 kHz (0-10 dB) but was intermediate between the control and pre treatment groups at 4 and 8 kHz (23 dB). Animals pre-treated with antioxidant had a significant reduction in hair cell loss but those post-treated with antioxidant had no protection from hair cell loss. These findings demonstrate the feasibility of reduction of noise-induced hearing loss using clinically available antioxidant compounds. PMID- 11033254 TI - Gastric proton pump is expressed in the inner ear and choroid plexus of the rat. AB - Inner ear fluids and cerebrospinal fluid show remarkably stable ionic concentrations, particularly that of K(+) and H(+), but the mechanisms which control the homeostasis of these media are not well understood. We investigated a possible role of the gastric H, K-ATPase (gH,K-ATPase) pump in this control since this pump is known to be expressed in other tissues than gastric parietal cells. Here, we show by reverse transcription-polymerase chain reaction that the rat gH,K-ATPase alpha- and beta-subunits are expressed in the inner ear (lateral wall, organ of Corti and spiral ganglion cells), while only the alpha-subunit is expressed in the choroid plexus (CP). The presence of the alpha-subunit in the inner ear and CP was confirmed by immunoblotting. Immunohistochemistry localized this protein in the intermediate cells of the stria vascularis, in the spiral ligament and the spiral ganglion. gH,K-ATPase could be involved in the maintenance of H(+) and K(+) equilibria in cerebrospinal and labyrinthine fluids. PMID- 11033255 TI - The responses of single units in the ventral cochlear nucleus of the guinea pig to damped and ramped sinusoids. AB - Human listeners hear an asymmetry in the perception of damped and ramped sinusoids; the partial loudness of the envelope component is greater than the partial loudness of the carrier component for damped sinusoids. Here we show that an asymmetry also occurs in the physiological responses of most units in the ventral cochlear nucleus to these same sounds. The activity elicited by damped sinusoids is mainly restricted to the beginning of each envelope period, which is not the case for ramped sinusoids. This can be quantified by computing the ratio of the tallest bin of the modulation period histogram to the total number of spikes (the peak-to-total ratio, p/t). Damped sinusoids produce a higher p/t than ramped sinusoids, which demonstrates physiological temporal asymmetry. It is also the case that ramped sinusoids typically elicit more spikes than damped sinusoids. The physiological asymmetry occurs where the perceptual asymmetry is present. It is maximal at modulation half-lives of 4 and 16 ms, greatly reduced at 1 ms and absent at 64 ms. Different unit types exhibit differing degrees of temporal asymmetry. Onset units produce the greatest p/t asymmetry, primary-like units produce the least asymmetry and chopper units are in-between. With regard to total spike count, the maximal asymmetry occurs with chopper units. If primary like units are assumed to reflect the activity in primary auditory nerve fibres, then there is enhancement of temporal asymmetry in the ventral cochlear nucleus by both onset and chopper units. PMID- 11033256 TI - Morphine inhibits an alpha9-acetylcholine nicotinic receptor-mediated response by a mechanism which does not involve opioid receptors. AB - Nicotinic acetylcholine (nACh) receptors are known to be targets for modulation by a number of substances, including the opiates. It is known that acetylcholine (ACh) coexists with opioid peptides in cochlear efferent neurons, and such a colocalization has been proposed for the vestibular system. In the present study we test the hypothesis that morphine, an opioid receptor agonist with a broad spectrum of selectivity, modulates alpha9nACh receptor-mediated responses in frog vestibular hair cells. Morphine dose-dependently and reversibly inhibited ACh induced currents as recorded by the perforated patch-clamp method. In the presence of morphine the ACh dose-response curve was shifted to the right in a parallel fashion, suggesting a competitive interaction. However, naloxone did not antagonize the inhibition produced by morphine. To test the hypothesis that morphine could interact with the alpha9nACh receptor without the involvement of opioid receptors, experiments were performed using Xenopus laevis oocytes injected with the alpha9nACh receptor cRNA. The currents activated by ACh in Xenopus oocytes, a system that lacks opioid receptors, were also dose-dependently inhibited by morphine. We conclude that morphine inhibits the alpha9nACh receptor mediated response in hair cells and Xenopus oocytes through a mechanism which does not involve opioid receptors but may be a direct block of the alpha9nACh receptor. PMID- 11033257 TI - Neuronal death, not axonal degeneration, results in significant gliosis within the cochlear nucleus of adult chickens. AB - Injury to the central nervous system initiates a series of events that leads to neuronal cell death and glial activation. Astrocytes respond to damage and disease by becoming hyperplastic and hypertrophied. This 'reactive gliosis' is also accompanied by the upregulation of the intermediate filament protein glial fibrillary acidic protein, the release of growth factors and the formation of the glial scar. However, the signaling cascades which regulate these events, and the molecular mechanisms that give rise to this diverse response, have not been fully elucidated. For example, the role played by degenerating neurons vs. degenerating axons in the activation of astrocytes remains to be determined. To investigate the influence of neuronal cell death vs. axonal degeneration on gliosis, the current study examines the astrocyte response to cochlea removal in two different breeds of adult chickens, one of which exhibits neuronal cell death within the brainstem nucleus magnocellularis (NM) following the lesion and one which does not. Our results indicate that degeneration of NM neurons leads to large increases in both glial proliferation and hypertrophy, while eighth nerve degeneration without NM cell death results in very small increases in glial proliferation. PMID- 11033258 TI - Acute changes in cochlear potentials due to cisplatin. AB - The purpose of this study was to investigate how the hair cells and stria vascularis are affected at the onset of cisplatin ototoxicity. The effects on the endocochlear potential (EP) and the cochlear microphonics (CM) were observed simultaneously in two groups of adult chinchillas receiving as follows: (1) 5 microl of cisplatin (1 mg/ml) in normal saline, and (2) 5 microl of normal saline on the round window. The EP and the CM were recorded for 12-14 h after cisplatin application, and morphological changes were assessed using scanning electron microscopy. Both the EP and the CM amplitude demonstrated a profound reduction, and a very strong correlation was observed between these two values during this time period. Although the reduction of the EP and the CM was observed by 12-14 h, only very slight degeneration of outer hair cells was seen at that time. These data suggested that a reduction of the EP which was caused by the alteration of the stria vascularis might be primarily responsible for very early changes in cochlear function after topical cisplatin application, while later changes were the direct result of hair cell damage. PMID- 11033259 TI - Modelling convergent input onto interaural-delay-sensitive inferior colliculus neurones. AB - Convergent input from cells in the medial superior olive (MSO) and lateral superior olive (LSO) onto a single inferior colliculus (IC) cell explains many findings that are not compatible with a simple coincidence detector mechanism. Here this explanation is tested using a physiologically accurate computer model of the binaural pathway in which the input to the IC cell is either from two MSO cells or a MSO and a LSO cell. Auditory nerve (AN) spike trains are formed by a stochastic hair cell model following a basilar membrane simulation using a gammatone filter. In subsequent cells input spikes cause post-synaptic potentials (PSPs) which are summed causing the cell to fire when the sum crosses a threshold. The individual cells are matched to the physiology by varying the number of inputs, the magnitude and duration of the PSPs and the firing threshold. Non-linear best-phase-versus-frequency functions arise if the two IC inputs have different best frequencies and different characteristic delays. One input can be selectively suppressed by turning on an additional tone at the worst phase of that input. Non-zero characteristic phases arise if the characteristic frequencies of the AN fibres feeding into a single superior olive cell are mismatched. PMID- 11033260 TI - Sodium pentobarbital abolishes bursting spontaneous activity of dorsal cochlear nucleus in rat brain slices. AB - There is evidence that pentobarbital, a commonly used anesthetic, can affect neuronal activity, but its effects on particular neurons of the dorsal cochlear nucleus (DCN) are not well known. Bursting (complex spiking) spontaneous activity has been observed in the DCN in brain slice preparations and in recordings from unanesthetized decerebrate animals, but seldom in experiments with anesthetized animals. This study investigated the effects of pentobarbital on spontaneous activity in the DCN in brain slices. Most extracellularly recorded bursting neurons decreased firing rates and reversibly changed their firing to simple spiking with irregular intervals during pentobarbital. Some reversibly stopped firing after the change to an irregular pattern. Most neurons with regular spontaneous activity (simple spiking) showed decreased firing rates and more irregular intervals during pentobarbital. The results also suggest some involvement of gamma-aminobutyric acid type A receptors in the pentobarbital effects. PMID- 11033261 TI - Chronic microstimulation in the feline ventral cochlear nucleus: physiologic and histologic effects. AB - This study was conducted to help to establish the feasibility of a multi-channel auditory prosthesis based on microstimulation within the human ventral cochlear nucleus, and to define the range of stimulus parameters that can be used safely with such a device. We chronically implanted activated iridium microelectrodes into the feline ventral cochlear nucleus and, beginning 80-250 days after implantation, they were pulsed for 7 h/day, on up to 21 successive days. The stimulus was charge-balanced pulses whose amplitude was modulated by a simulated human voice. The pulse rate (250 Hz/electrode) and the maximum pulse amplitude were selected as those that are likely to provide a patient with useful auditory percepts. The changes in neuronal responses during the multi-day stimulation regimens were partitioned into long-lasting, stimulation-induced depression of neuronal excitability (SIDNE), and short-acting neuronal refractivity (SANR). Both SIDNE and SANR were quantified from the changes in the growth functions of the evoked potentials recorded in the inferior colliculus. All of the stimulation regimens that we tested induced measurable SIDNE and SANR. The combined effect of SIDNE and the superimposed SANR is to depress the neuronal response near threshold, and thereby, to depress the population response over the entire amplitude range of the stimulus pulses. SIDNE and SANR may cause the greatest degradation of the performance of a clinical device at the low end of the amplitude range, and this may represent an inherent limitation of this type of spatially localized, high-rate neuronal stimulation. We determined sets of stimulus parameters which preserved most of the dynamic range of the neuronal response, when using either long (150 micros/phase) or short (40 micros/phase) stimulus pulses. Increasing the amplitude of the stimulus was relatively ineffective as a means of increasing the dynamic range of neuronal response, since the greater stimulus amplitude induced more SIDNE. All of the pulsed and unpulsed electrode sites were examined histologically, and no neuronal changes attributable to the stimulation were detected. There was some aggregation of glial cells immediately adjacent to some of the electrodes that were pulsed with the short-duration pulses, and at the highest current densities. PMID- 11033262 TI - Vulnerability to noise-induced hearing loss in 'middle-aged' and young adult mice: a dose-response approach in CBA, C57BL, and BALB inbred strains. AB - Vulnerability of the cochlea to noise-induced permanent threshold shifts (NIPTS) was examined in young adult (1-2 months) and 'middle-aged' (5-7 months) CBA/CaJ, C57BL/6J, and BALB/cJ inbred mice. For each age and strain, a dose-response paradigm was applied, whereby groups of up to 12 animals were exposed to intense broadband noise (110 dB SPL) for varying durations. Exposure durations reliably associated with <10% and >90% probability of a criterion amount of NIPTS (determined 2 weeks post-exposure) were identified, and the minimum NIPTS exposure and the slope of the dose-response relation were then derived by numerical modeling. For all three strains, young adult mice were more susceptible to NIPTS than older adults; That is, a shorter exposure was able to cause NIPTS in the younger mice. Strain comparisons revealed that C57 mice were more susceptible than CBAs in the older age group only. At both ages examined, however, BALB mice were most susceptible to NIPTS. When animals with a similar amount of NIPTS were compared, outer hair cell loss in the cochlear base was more widespread in the younger animals. BALB mice appear particularly susceptible to noise-induced outer hair cell loss throughout life. Our data suggest that the mechanism or site of noise injury differs between young adults and older adults, and may depend on genetic background. The finding that both BALB and C57 mice, which show pronounced age-related hearing loss, are also especially vulnerable to noise supports the notion that genes associated with age-related hearing loss often act by rendering the cochlea susceptible to insults. PMID- 11033263 TI - Measurement of stapedius contraction during vocalization effort in patients after laryngectomy or tracheostomy. AB - The contraction of the stapedius muscle during the effort of vocalization was examined by measurement of acoustic compliance in subjects who had undergone laryngectomy or tracheostomy. No significant level of persistent compliance change was recorded in any of the subjects, indicating the absence of effective contraction of the stapedius during the vocalization effort. In two subjects who use an electrolarynx, although no significant compliance change resulted from the simple vocalization effort without the electrolarynx (no actual vocalized sound), a remarkable level of persistent compliance change was observed during the vocalization using the electrolarynx. These results seem to indicate that a simple vocalization effort without actual voice cannot elicit effective contraction of the middle ear muscles, and that sound generation during vocalization is essential for effective contraction of the middle ear muscles during vocalization, at least in some human subjects. PMID- 11033264 TI - The prevalence of Helicobacter pylori: are emergency medicine residents at risk?. AB - Helicobacter pylori is strongly associated with peptic ulcer disease and gastric cancer. It is a cause of chronic active gastritis. Our hypothesis was that Emergency Physicians might be at increased risk for infection due to their exposure to gastric contents from procedures performed in the Emergency Department. Our goal was to test a group of Emergency Physicians for H. pylori infection. In this cross sectional study, we tested a group of volunteers from an Emergency Medicine residency program for H. pylori infection by serum analysis for IgG. A matched control group was also tested. Forty-five subjects were enrolled; one subject's sample was lost. Three of 44 subjects (6.8%) were positive for H. pylori antibodies. Seven of 44 controls (15.9%) tested positive. Thus, Emergency Physicians do not have increased H. pylori infection rates and may be at less risk than the general population. Routine testing is not recommended. PMID- 11033265 TI - Bedside measurement of D-dimer in the identification of bacteremia in the emergency department. AB - The objective of this pilot study was to determine the clinical utility of the SimpliRed D-dimer bedside assay to identify patients with bacteremia in a university hospital Emergency Department. We tested 265 patients and compared blood culture results with a novel D-dimer semiquantitative whole blood assay. Bacteremia was confirmed in 25/262 patients. Sensitivity of D-dimer assay was 66.7% for Gram-positive bacteremia and 61.5% for Gram-negative bacteremia with negative predictive value of 98% for Gram-positive and 96% for Gram-negative bacteremia patients. Measurement of D-dimer appears to be of value in identifying patients at low risk for bacteremia and can be accomplished rapidly using a whole blood semiquantitative bedside assay. Although increases in D-dimer are not detected in all patients subsequently documented to have bacteremia on a single sampling, the results of this and other earlier studies suggest assay of D-dimer is useful in rapid differentiation of patients with bacteremia from those who have no bacteremia using blood culture positivity as the standard for bacteremia. PMID- 11033267 TI - Exposure to HIV: medical management and legal implications. AB - The management of exposures to HIV, whether occurring in the occupational or non occupational setting, involves balancing the risk and inconvenience of antiviral therapy that lacks strong evidence of efficacy against the benefit of it possibly preventing a potentially fatal disease. Clinicians increasingly will be responsible for managing Health Care Workers (HCWs) and other persons exposed to HIV. This will require making a clinical assessment of the risk of HIV transmission and making recommendations for postexposure prophylaxis (PEP) according to the CDC, and other guidelines. Management also requires applying the elements of the doctrine of informed consent, considering involuntary testing of source patients, and reporting of exposure incidents. Proper management will protect all those involved: the exposed person will be protected from unacceptable risks either of contracting HIV, should PEP be indicated, or harm caused by PEP when it is not; the source patient will be protected from unconstitutional invasions of privacy. Finally, the clinician will be protected from claims of malpractice as a result of management of an HIV exposure. PMID- 11033266 TI - Management of patients with suspected deep vein thrombosis in the emergency department: combining use of a clinical diagnosis model with D-dimer testing. AB - The management of patients presenting to hospital Emergency Departments with suspected deep vein thrombosis is problematic since urgent diagnostic imaging is at times unavailable. We evaluated the accuracy of a rapidly available D-dimer test and the potential of combining D-dimer testing with an explicit clinical model to improve the management of patients with suspected deep vein thrombosis. Two hundred and fourteen patients with suspected deep vein thrombosis presenting to the Emergency Departments of two tertiary care institutions were enrolled in this prospective cohort study. Patients were evaluated by an Emergency Physician who determined the pre-test probability for deep vein thrombosis to be either low, moderate, or high using an explicit clinical model. Patients were managed according to their pre-test probability category by specific algorithms that in all cases included venous ultrasound imaging within 24 h and a 90-day follow-up for the development of thromboembolic complications. Patients also underwent fingerstick SimpliRED(R) whole blood agglutination D-dimer testing; however, D dimer results did not influence subsequent patient management. D-dimer had a sensitivity of 82.5% and a specificity of 84.9% for the diagnosis of deep vein thrombosis. The observed negative predictive value of D-dimer was 96.9% (95% CI, 93.0% to 99.1%) overall, and 100% (95% CI, 96.3% to 100%) in low probability patients, 94.1% (95% CI, 83.8% to 98.8%) in moderate probability patients, and 86.7% (95% CI, 59.4% to 98.3%) in high probability patients. SimpliRED(R) D-dimer has a high negative predictive value and may be useful in excluding the diagnosis in patients at low pre-test probability for deep vein thrombosis. PMID- 11033268 TI - Retention of urethrovesical foreign bodies: case report and literature review. AB - Insertion of foreign bodies into the urethram to obtain sexual pleasure may be complicated by their passage into the urinary bladder along with an inability for the patient to recover the foreign body. We present such a case, review the relevant literature, and discuss Emergency Department (ED) diagnosis and management. PMID- 11033269 TI - Chronic paronychia, osteomyelitis, and paravertebral abscess in a child with blastomycosis. AB - Blastomycosis is an unusual fungal infection in children. It is often a chronic infection characterized by granulomatous and suppurative lesions. Clinical manifestations include either pulmonary findings or disseminated disease. Disseminated blastomycosis usually begins with a lung infection that spreads to the skin, bones, and central nervous system. This is a case report of a child with chronic blastomycosis presenting with chronic paronychia, fever, cough, malaise, and back pain. The child underwent surgical drainage of a paravertebral abscess and administration of intravenous amphotericin B. He was discharged in good condition on oral therapy with ketoconazole. The literature on blastomycosis, with particular emphasis on clinical presentations and management, is reviewed. When the history and physical examination suggest a chronic granulomatous or disseminated disease, such as tuberculosis, the physician must include blastomycosis in the differential. PMID- 11033270 TI - Chronic ectopic pregnancy--two cases of acute rupture despite negative beta hCG. AB - We present the cases of two women with chronic ectopic pregnancies who presented with acute tubal rupture and hemoperitoneum despite negative beta-human chorionic gonadotropin (beta hCG) pregnancy tests. The appropriate screening use of beta hCG assays to ascertain pregnancy status, the mechanisms by which ectopic pregnancy may be seen with negative beta hCG tests, and the limitations of various assays are discussed. One patient, not initially believed to be pregnant, underwent computed tomography (CT) scan. Experience with use of CT scan in ectopic pregnancy diagnosis is limited; our case illustrates some of the possible CT scan findings. These cases illustrate the potential for ectopic pregnancy to rupture with low, if not undetectable beta hCG hormone levels, and consequently why it is not recommended to rely on quantitative beta hCG levels to guide the decision to proceed with ultrasound imaging. PMID- 11033271 TI - Retrospective analysis of ingestions of iron containing products in the united states: are there differences between chewable vitamins and adult preparations? AB - Iron is the one of the leading causes of pediatric poisoning deaths in the United States. Most cases of serious iron overdose reported in the medical literature have resulted from adult formulations of iron. To begin evaluating the possibility that differences in toxicity exist between iron preparations, we performed a retrospective evaluation of all exposures to pediatric and adult iron products reported to the American Association of Poison Control Centers' (AAPCC) Toxic Exposure Surveillance System (TESS) from 1983 to 1998. We attempted to determine the incidence of fatal iron poisoning for each group. A total of 195,780 ingestions of children's vitamins containing iron were reported to the TESS between 1983 and 1998 with no resulting fatalities. During the same twelve year study period, 147,079 exposures to adult forms of iron were reported with 60 fatalities (p < 0.0001). A prospective study is required to assess whether differences may exist in the toxicity of these two iron preparations. PMID- 11033272 TI - Rapid sequence intubation in the field versus hospital in trauma patients. AB - We conducted a retrospective review of all adult trauma patients who underwent prehospital field rapid sequence intubation (RSI) by aeromedical crews from 1988 through 1995 and compared them to all trauma patients who arrived by ground transportation and underwent RSI in the trauma suite from 1992 through 1995 at a University hospital. Of the 47 field RSI patients, 46 (97.9%) were successfully intubated, whereas 263 of the 267 (98.5%) hospital RSI patients were successfully intubated. There were no statistical differences in success rates, number of attempts, or immediate intubation events in the procedure between the two groups. There were no differences in delayed events with the exception of pneumonia, which occurred more frequently in the field RSI group (28% vs. 6%, respectively). We performed a subgroup analysis on isolated head injury patients to evaluate outcome. There was no difference in total hospital days, length of ICU stay, mortality or final disposition in the two head injury groups. Though this study is limited by small sample size, we conclude that field RSI is equally successful and safe as hospital RSI. PMID- 11033273 TI - Salter-Harris I fractures of the distal radius misdiagnosed as wrist sprain. AB - Over a period of 5 years, 65 children under 12 years of age were diagnosed as having "wrist sprain" at the American University of Beirut Medical Center, Emergency Department. Thirty-eight children were reevaluated; 33 patients (86.8%) were diagnosed as having a Salter-Harris I fracture of the distal radius that necessitated further treatment to relieve symptoms and allow early wrist function. The diagnosis of "wrist sprain" is often inadequate in establishing a proper treatment regimen. Rather, by taking a careful history, performing an exacting examination, and utilizing appropriate diagnostic aids, an accurate diagnosis of wrist injury can be established. PMID- 11033274 TI - Altered mental status in alcoholism. PMID- 11033275 TI - The eyes see only what the mind knows. PMID- 11033276 TI - Total lung collapse. PMID- 11033277 TI - Technical improvisation in emergency medicine. PMID- 11033278 TI - The quest for the cure of all chronic illnesses: a wake-up call from the President of the United States. PMID- 11033280 TI - Ten questions that work. PMID- 11033279 TI - The compensatory pause: declining emergency physician income. PMID- 11033281 TI - Long-term follow-up of vitamin B(6)-responsive West syndrome. AB - We performed a clinical and electroencephalographic follow-up study on 25 patients with West syndrome that was responsive to vitamin B(6) (eight cryptogenic patients and 17 symptomatic patients) who were older than 3 years at the last follow-up. All cryptogenic patients and 13 symptomatic patients were seizure free at the last follow-up. All cryptogenic patients and seven symptomatic patients had intelligent quotient or developmental quotient scores of 75 or higher. The recurrence of clinical seizures was always associated with increases in epileptic discharges. We could successfully discontinue pyridoxal phosphate administration in four cryptogenic and four symptomatic patients who were 1 year, 8 months to 24 years old. PMID- 11033282 TI - CAT/CLAMS: its use in detecting early childhood cognitive impairment. AB - The Cognitive Adaptive Test/Clinical Linguistic and Auditory Milestone Scale (CAT/CLAMS), a neurodevelopmental tool for the cognitive assessment of infants and toddlers, correlates well with the Bayley Scales of Infant Development. In 1993 the Bayley Scales were revised and the second edition published (BSID-II). This study was designed to determine how well the CAT/CLAMS correlates with the BSID-II and its utility in identifying mild and severe cognitive impairment. Sixty-eight infants and toddlers (age range = 14-48 months), referred for suspected developmental delays, were administered the CAT/CLAMS and BSID-II and the results compared. The correlation between the two instruments was strong (r = 0.89, P<0.0001). The CAT/CLAMS was sensitive (81%) and specific (85%) for detecting overall cognitive impairment (BSID-II less than 70) and was even more sensitive (100%) and specific (96%) in detecting severe cognitive impairment (BSID-II less than 50). The physician using the CAT/CLAMS formulated a clinical impression of cognitive impairment that was sensitive (95%) and specific (84%) compared with formal psychologic testing. The CAT/CLAMS correlates well with the BSID-II. It is useful for detecting and quantifying mild and severe cognitive impairment. It permits the physician to formulate an accurate clinical impression of cognitive impairment consistent with possible mental retardation. PMID- 11033283 TI - Regional specificity of localized cortical lesions in West syndrome. AB - West syndrome, although classified as a generalized epilepsy, is associated with localized cerebral lesions in some cases. However, similar localized cortical abnormalities usually can result in partial epilepsy, instead of West syndrome. We performed this study to determine the additional factors that result in West syndrome instead of partial epilepsy in patients with localized cerebral lesions. We reviewed the pathologic features, topographic localization, and side of unilaterally defined cerebral lesions in relation to the seizure types in 39 epileptic patients, including five patients who presented with West syndrome. The lesions of all five patients with West syndrome involved the temporal or occipital lobes (or both). Among the nine with an occipital lesion, four had West syndrome. In the 19 with a temporal lesion, three had West syndrome. However, in the 16 patients with a frontal lesion, none exhibited epileptic spasms. In four of the five with West syndrome the lesions were on the right side; 23 had lesions on the right, 16 had them on the left. Temporo-occipital lesions and lesions on the right were related to the genesis of West syndrome, which would be in close correlation with normal brain maturation. PMID- 11033285 TI - Melatonin for the treatment of handicapped children with severe sleep disorders. AB - Sleep disorders are common in children with mental retardation and neurologic disorders. Melatonin, a recently developed natural compound, has been used successfully in sleep disorders. I report my experience with melatonin in an open, prospective trial to treat circadian rhythm sleep disorder in handicapped children. The sleep disorder had been present for at least 6 months and had not responded to at least one hypnotic drug. The therapeutic response was recorded according to the average number of hours asleep per 24 hours, average number of awakening per night, average number of nights with delayed sleep onset, and average number of nights with early morning arousals. Ten consecutive children (four males, six females; age range = 1-11 years, mean 5.4) were included. Nine children had documented mental retardation that was severe in six (67%). Most had epilepsy and visual impairment (70%). All children were monitored for 4-12 months (mean 7.5 months) after the initiation of 3-mg bedtime melatonin. Most (80%) had a dramatic response to melatonin. No side effects were reported. Melatonin is a well-tolerated, safe, relatively inexpensive, and effective drug, with minimal side effects, for the treatment of severe circadian rhythm sleep disorder in handicapped children. Wider use of this drug is recommended. PMID- 11033284 TI - Melatonin improves sleep-wake patterns in psychomotor retarded children. AB - Five children with severe psychomotor retardation (mean age 8.2+/-3.6 years) and irregular sleep-wake patterns underwent 1 week of wrist actigraphic monitoring before and after treatment with 3 mg melatonin. Three underwent multiple measurements of urinary sulfatoxymelatonin levels. Urine sulfatoxymelatonin levels were abnormally low, without any significant day/night differences. Melatonin treatment increased nighttime sleep from 5.9+/-0.8 to 7.3+/-0.5 hours (paired t test, P<0.01) and sleep efficiency from 69.3%+/-6.2% to 88.3%+/-2.3% (P<0.01). Daytime sleep decreased from 3.2+/- 1.2 to 1.7+/-1.2 hours (P<0.05). Thus, no change in 24-hour total sleep time (9.1+/-1.5 vs. 9.0+/-1.6 hours) occurred. Administration of 3 mg melatonin to five severely psychomotor retarded children resulted in a significant improvement in their sleep-wake patterns. PMID- 11033286 TI - West syndrome in tuberous sclerosis complex. AB - West syndrome occurs commonly in children with tuberous sclerosis complex and is associated with a grave prognosis for cognitive and seizure outcomes. We sought to determine the epilepsy outcome of children with tuberous sclerosis complex and West syndrome and whether EEG, MRI, or steroid therapy duration were different in those whose epilepsy improved compared with those with intractable seizures. Seventeen patients with tuberous sclerosis complex and West syndrome were identified. For each patient, two sets of clinical evaluations, EEG and MRI data, and treatment information separated by at least 12 months were obtained. The patients were divided into two seizure outcome groups. EEG, MRI, and treatment data were compared between the groups. The intellectual deficiency was either severe (76%) or moderate (24%). Seizure control improved in 10 and worsened in seven, without mortality (follow-up range = 12-216 months). No significant differences in EEG background, MRI findings, or steroid treatment duration were evident between the groups. The difference in EEG-sleep approached statistical significance (P = 0.06). Our findings did not confirm reports of high mortality and poor epilepsy outcome in intellectually deficient children with West syndrome and tuberous sclerosis complex. EEG sleep was the best indicator of seizure control and approached statistical significance. The duration of steroid therapy had no influence on seizure control. PMID- 11033287 TI - Grading of cerebral autoregulation in preterm and term neonates. AB - The response of cerebral blood flow velocity to a single spontaneous transient rise in blood pressure was studied to grade the cerebral autoregulatory response of newborns. Blood pressure was measured continuously through an umbilical or peripheral arterial catheter; continuous flow velocity recordings were taken from the middle cerebral artery using continuous wave Doppler ultrasound. From a cohort of 62 healthy term and preterm neonates, 325 transients in mean arterial blood pressure and mean cerebral blood flow velocity were identified for analysis using a foot-seeking algorithm. An initial classification of active or impaired autoregulation was given to each transient using a self-clustering technique. The grading of the transients was studied by examining the slope of the return of the cerebral blood flow velocity to baseline. Negative slopes indicate a normal autoregulation; slopes of 0 or greater indicate an absence of autoregulation. This classification was in agreement with the self-clustering method (Cohen's kappa = 0.94, P<0.0001). The relationship between the autoregulatory response assessed by the grading method and gestational age, postnatal age, and PCO(2) was examined using linear regression analysis. A significant relationship with gestational age (P = 0.002) but not PCO(2) (P = 0.06) or postnatal age (P = 0.14) was evident. PMID- 11033288 TI - Topographical features of the sensory-evoked responses in malformed brains. AB - To reveal the functional organization of the somatosensory area in the dysgenetic cortex, somatosensory-evoked potentials were examined in seven patients with congenital brain anomalies diagnosed by magnetic resonance imaging, including six patients in whom multichannel recordings over the scalp were used. In four patients with polymicrogyria/pachygyria and two with lissencephaly, the early cortical responses, frontal P20 and parietal N20, were absent in the cortex contralateral to the stimulated side. The first cortical response was a positive wave that appeared predominantly over the centroparietal area in five patients, and in the frontal area in the other patient with polymicrogyria/pachygyria. These findings suggest that the differentiated somatosensory function is distributed normally in the centroparietal cortex in most cases of widespread cortical dysplasia. However, the absence of P20/N20 may indicate a hypoplastic central sulcus or functionally undifferentiated subdivision of the somatosensory cortex in these patients. The absence of cortical responses in the patient with holoprosencephaly may correspond with growth failure of the thalamocortical afferent projections in this disorder. PMID- 11033289 TI - Intracerebral aneurysms in human immunodeficiency virus infection: case report and literature review. AB - We describe a child with human immunodeficiency virus infection who presented with a large subarachnoid hemorrhage. She had multiple saccular and fusiform aneurysms in the proximal cerebral arterial circulation and no evidence of bacterial or fungal infection. The arteriopathy coincided with a high human immunodeficiency virus RNA load. Human immunodeficiency virus may cause cerebral arteriopathy with potentially life-threatening complications. PMID- 11033290 TI - Visual field plasticity in a female with right occipital cortical dysplasia. AB - Brain plasticity refers to its ability to recover after damage. Visual field plasticity is not well recognized. We report a 12-year-old female who first presented with recurrent seizures and was subsequently found to have a large, right occipital cortical dysplasia on magnetic resonance imaging. Her visual field by Goldmann perimetry was totally normal. Visual-evoked potential studies revealed the left hemifield P100 response was detected maximally at the right temporal and parietal regions. A weak but reproducible right hemifield P100 response was located at the right medial skull base. Functional magnetic resonance imaging with flashlight stimulation revealed cerebral activity mainly at the right posterior temporal and parietal lobes and left occipital lobe. These studies suggested that the left hemifield function was located at the right posterior temporal and parietal lobes. The left occipital lobe may also have been reorganized, with a P100 vector pointing out from its inferiomedial base. We reviewed other related reported cases. We believe that visual-evoked potential studies and visual functional magnetic resonance imaging should be performed more liberally for recognition of visual field plasticity. PMID- 11033291 TI - Static magnetic field therapy for pain in the abdomen and genitals. AB - Two adolescents with debilitating, medication-resistant, chronic pain of the low back and abdomen with intermittent pain of the genitalia were diagnosed with intervertebral disk disease at spinal cord levels that correlated with their signs. Both patients had undergone multiple evaluations by physicians of different specialties and both underwent appendectomy without relief of their pain. The history of the onset of pain was important in determining the affected levels. The pain of both individuals was mimicked and localized by percussion of the vertebral spines at the level of disk protrusion. This maneuver and careful review of the history were important in making the correct diagnosis in each case. In both patients, treatment with novel magnetic devices provided rapid relief that was sustained for more than 2 years. These cases highlight the need for careful evaluation and correct diagnosis of abdominal and genital pain in young patients to avoid costly and unnecessary medical intervention and the stigma of painful debility. PMID- 11033292 TI - A case of atypical absence seizures induced by leuprolide acetate. AB - We report a case of a 13-year-old female with atypical absence seizures induced by prolonged administration of long-acting leuprolide acetate (LA). This patient had brain involvement resulting from chemotherapy and radiotherapy for a medulloblastoma. At 13 years of age, administration of long-acting LA was started. After the third dose of long-acting LA, atypical absence seizures appeared. After discontinuing long-acting LA, the seizures stopped without administration of any antiepileptic drugs. However, 2 years, 6 months later, the same seizures again appeared. On the basis of the findings of endocrinologic investigations and the reported data of pharmacokinetics of LA, we speculate that her seizures were induced by LA and that the seizures were associated with the presence of brain damage in the patient. Care should therefore be taken when using long-acting LA or other gonadotropin-releasing hormone analogues for pediatric patients with diffuse brain damage. PMID- 11033293 TI - Reversible deafness caused by biotinidase deficiency. AB - We present a child with complete biotinidase deficiency who developed bilateral sensorineural deafness without a response to a maximal stimulus of 90 dB in brainstem acoustic-evoked response. After treatment with 20 mg biotin daily, a repeated brainstem acoustic-evoked response demonstrated an improved hearing threshold of 65 dB, and the child began to talk. The case is a rare example of reversible hearing loss caused by to biotinidase deficiency and highlights the need for immediate replacement therapy once the diagnosis is established. PMID- 11033294 TI - Prolonged generalized epileptic seizures triggered by breath-holding spells. AB - We report a 3-year-old female with anoxic-epileptic seizures. Beginning at 11 months of age, she had repeated breath-holding spells with transition into generalized tonic-clonic seizures or status epilepticus. Interictal electroencephalography exhibited no abnormalities. A multidisciplinary diagnostic approach revealed a severely disturbed mother-daughter relationship that was the trigger of the breath-holding spells. Psychotherapy for the mother and daughter led to cessation of the breath-holding spells and, consequently, of the anoxic epileptic seizures. Her further development was largely normal. We discuss the etiology and treatment of anoxic-epileptic seizures. This case is the first reported case of anoxic epileptic seizures that responded to psychologic rather than antiepileptic treatment. We advocate an initial psychologic assessment to help determine the appropriate treatment in children with recurrent anoxic epileptic seizures. PMID- 11033295 TI - Successful zonisamide treatment for infants with hypsarrhythmia. AB - We determined that zonisamide was effective in three epileptic infants with hypsarrhythmia. Two patients had the electroencephalographic pattern of hypsarrhythmia associated with developmental delay but lacked the characteristic tonic spasms. A third patient exhibited the typical triad of infantile spasms but had a poor general condition requiring controlled ventilation. We therefore chose zonisamide instead of adrenocorticotropin for these patients. Our data suggest that zonisamide may be a useful treatment in patients falling short of the diagnostic triad of infantile spasms or complicated by a poor overall medical condition. PMID- 11033296 TI - Contribution of three-dimensional culture to cancer research. PMID- 11033297 TI - Spheroids and cell survival. AB - In contrast to most traditional cell culture systems, spheroids represent a unique opportunity to recapitulate aspects of cell homeostasis and as such better reflect in vivo tumor biology. This review highlights recent spheroid-based studies which have defined a role for intercellular adhesion in dictating cellular fate. Critical roles for adhesion molecules, such as the integrin and cadherin families, have been established. In addition, it appears that components of the signaling pathways that control cellular proliferation may also influence the cellular decision to live or die. Further complexity arises from the action of soluble factors, like epidermal growth factor, and three-dimensional culture systems are providing insights into how integration of these multiple signals is achieved. Given that cellular adhesion may serve as a regulator of apoptosis, and in particular that correct receptor engagement can activate signaling pathways promoting cell survival, additional implications for our understanding of tumor biology are discussed. PMID- 11033298 TI - Apoptosis, cell adhesion and the extracellular matrix in the three-dimensional growth of multicellular tumor spheroids. AB - In the last few years, it has become increasingly apparent that cell survival and death, especially apoptosis, strongly depend on cell adhesion and the extracellular matrix. In addition, it has also become clear that the use of three dimensional multicellular tumor spheroids, which mimick more closely solid tumors in vivo, are a realistic experimental model to investigate many aspects of tumor biology. In the present review, after a general overview of the current knowledge regarding apoptosis, cell adhesion and the extracellular matrix, the results obtained utilizing multicellular tumor spheroids in these types of studies are discussed. The main conclusion that may be drawn from a synthesis of the literature on these topics is that investigations with multicellular tumor spheroids yield much useful information that is sometimes in contradiction to that obtained with monolayer cultures, but is closer to that derived from in vivo studies. Consequently, the authors encourage that these three-dimensional systems be used in many studies in which cell death and adhesion are being examined. PMID- 11033300 TI - Spheroids: relation between tumour and endothelial cells. AB - It has recently been established that the microenvironment plays a major role in many physiological and pathological events. Indeed cell-cell and cell extracellular matrix contacts are necessary for much cellular function such as differentiation, proliferation, cell death, apoptosis and angiogenesis. For growth, proliferating tumour cells need to be fed by nutrients and oxygen brought by new vessels. In this context, scientists seek a new model that allows for the investigation of both angiogenesis and the influence of the microenvironment on this phenomenon. The purpose of this paper is to review the literature on the relation between tumour and endothelial cells grown as spheroids, a technique that allows us to study in three-dimensions the influence of cell contact on this growth. For the purpose of clarification, this review has recategorised the different studies on spheroids into three classes: (1) spheroids grown in vitro and then reimplanted in animals to follow endothelial cell infiltration; (2) spheroids grown in vitro and then cultured on endothelial cell monolayers; (3) tumours grown in vitro such as organotypic culture. This review attempts to demonstrate that spheroid cell cultures are useful for studying the relation between tumour and endothelial cells and to analyse physiological phenomena such as wound healing, extravasation and intravasation. PMID- 11033299 TI - Serum signaling factors and spheroids. AB - Normal epithelial cells grow as a sheet-like structure. Upon malignant transformation, epithelial cells grow as multicell aggregates. Adopted in tissue culture, most tumor cells revert to adherent monolayer. In tissue culture, as early as 1958, anchorage-independent multicellular spheroid cancer cells have been shown to revert to adherent monolayer in response to extracellular serum signaling factors. Such serum signaling factors have not yet been characterized. Recent studies reveal that the conversion of adherent monolayer to multicellular spheroids is also mediated by serum signaling factors such as carcinoembryonal antigen, interferon-gamma, insulin-like growth factor-II, heregulin beta1 and plasmin. The reports provide a new approach to investigate the regulatory system of tumor cell growth pattern as well as the effect of the change in growth pattern on various cellular functions. PMID- 11033301 TI - Autologous spheroid culture: a screening tool for human brain tumour invasion. AB - Spheroids are three-dimensional cell aggregates expressing histotypic organisation in vitro comparable to tissue continuity in vivo. They can be prepared from normal tissue and from tumour fragments. In the experiments presented here, dermal human spheroids and brain tumour spheroids are prepared from the same patient. The dermal tissue originates from the border of the incision wound made to effect a stereotactic brain tumour biopsy. The tumour originates from a fragment of the collected stereotactic biopsy. The dermal fragment and the brain biopsy are explanted in vitro to form confluent monolayers. At confluency, the dermal cells are transferred into small Erlenmeyer flasks and rotated at 37 degrees C for 1-2 days and rotation mediated spheroids are formed. Small flaps of the tumour monolayer are placed on a semisolid non adhesive substrate, reorganise and form agar overlay spheroids. After spheroid formation, a dermal spheroid is confronted with a brain tumour derived spheroid. The confronting pair, after adhering to each other, present an invasion model in vitro. The dermal spheroid functions as the autologous host for the brain tumour spheroid. Putative invasive cells present in the reaggregated brain spheroid will invade the dermal spheroid and destroy it. If no invasive cells are present in the tumour derived spheroid no morphologic changes will be seen in the dermal spheroid; 24 tested brain biopsy spheroids demonstrated a clear correlation between malignancy in situ and invasiveness in vitro. So it can be concluded that the autologous confrontation of brain tumour derived spheroids with dermal spheroids derived from the patient has a predictive value concerning malignant evolution and mimics the situation of the tumour in situ. PMID- 11033302 TI - Tumor biology and experimental therapeutics. AB - Recent research using multicellular tumor spheroids has resulted in new insights in the regulation of invasion and metastasis, angiogenesis and cell cycle kinetics. The onset and expansion of central necrosis in tumor spheroids has been characterized to be a complex interaction of several mechanisms; in a number of cases, necrosis is not a consequence of hypoxia or anoxia, but emerges as secondary necrosis following an accumulation of apoptosis in spheroids. Recent therapeutically oriented studies have been directed towards novel hypoxic markers, targeted therapy, multicellular-mediated drug resistance, and heavy ion irradiation of spheroids. Research efforts should be enhanced mainly in the fields of tumor tissue modeling by heterotypic three-dimensional (3D) cultures and of apoptotic versus necrotic cell death. Based on the fundamental differences between monolayer and 3D cultures, spheroids should become mandatory test systems in therapeutic screening programs. PMID- 11033303 TI - The multilayered postconfluent cell culture as a model for drug screening. AB - New drug development requires simple in vitro models that resemble the in vivo situation more in order to select active drugs against solid tumours and to decrease the use of experimental animals. In this paper, we review the characteristics and scope of a relatively simple cell-culture system with a three dimensional organisation pattern - the multilayered postconfluent cell culture model. Solid tumour cell lines from diverse origins when grown in V-bottomed microtiter plates reach confluence in 3-5 days and then start to form multilayers. The initial exponential growth of the culture is followed by a plateau phase when cells reach confluence. This produces changes in the morphology of the cells. For some cell lines, it is possible to observe cell differentiation. A substantial advantage of the system is the use of the sulforodamine B (SRB) assay to determine relative cell growth or viability, which allows semiautomation of the experiments. Several experiments were performed to assess the differences and similarities between cells cultured as monolayers and multilayers, and eventually, compared with the results for solid tumours and some other models such as spheroids. Cell-cycle analysis for multilayers showed a lower S-phase arrest, which is accompanied by a decrease in the expression of cell-cycle-related proteins and a decrease in cellular nucleotide pools. Gene and protein expression of topoisomerase I, topoisomerase II and thymidylate synthase expression were lower for multilayers, but no substantial changes were observed for the expression of DT-diaphorase. P53 expression increased. Multilayer cultures present distinctive properties for drug transport across the membrane, drug accumulation and retention. In fact, the transport of antifolates across the membrane, accumulation of topotecan and gemcitabine-triphosphate are reduced in multilayers when compared with monolayers, which may be related to a decrease in drug penetration to the inner regions of the multilayers. Alteration of these pharmacodynamic parameters is directly related to a decrease in drug activity. The most powerful application of multilayers is in the assessment of cytotoxicity. Solid tumour cell lines from different origins have been treated with several conventional and investigational anticancer drugs. The data show that multilayers are more resistant to the drugs than the corresponding monolayers, but there are substantial differences between the drugs depending on culture conditions, e.g. the difference was rather small for a drug such as cisplatin, miltefosine and EO9, a drug, which is activated under hypoxic conditions. Gemcitabine was active against ovarian cancer but not against colon cancer, resembling the in vivo situation. This observation was not evident with monolayer experiments. Another interesting application is the possibility to perform drug combination studies. The combination of gemcitabine and cisplatin proved to produce selective cell kill in H322 cells (non-small cell lung cancer cell line). Neither of the drugs was independently able to produce similar effects. In summary, multilayer cultures are relatively simple three-dimensional systems to study the effect of microenvironmental conditions on anticancer drug activity. The model might serve as a base for a more rigorous secondary in vitro screening. PMID- 11033304 TI - Effect of therapeutic macromolecules in spheroids. AB - Recent advances in biotechnology have allowed the production of new types of macromolecular therapeutic agents (antibodies, immunotoxins, cytokines, extracellular matrix molecule (ECM) proteins, vectors) that may eventually find broad clinical applications in the treatment of human tumors and other diseases. The model of the Multicellular Tumor Spheroids (MTS) represents a valuable tool to test the therapeutic potential of these new pharmacologic agents in a 3-D context. Specific questions pertaining to the behaviour in a 3-D setting of some of the macromolecules under evaluation for in vivo applications can also be addressed in the MTS model (e.g. 'binding site barrier', role of cell-cell and cell-ECM interactions). This paper reviews the most significant contributions regarding the delivery of macromolecules to MTS, the penetration and therapeutic effects of antibodies, radiolabelled antibodies, immunotoxins and other macromolecular compounds. PMID- 11033305 TI - Spheroids in radiobiology and photodynamic therapy. AB - Spheroids are tridimensional aggregates of tumor cells coming from one or several cell clones. This model, which mimics the micro-tumors structure and some of their properties, shows oxygen, pH and nutrient gradients inducing a necrotic area in the center of the spheroid. Analysis of spheroids, cultured under static or stirred conditions, can be performed on whole spheroids or dissociated spheroids. The spheroids sensitivity to ionizing radiation and photodynamic therapy can be altered by oxygen status, damage repair, intercellular commmunications and apoptosis induction, as in experimental tumor models. In radiobiology, the similarity of radiation response between spheroids and tumor xenograft bearing mice makes the spheroids to be a good alternative model to in vivo irradiation studies. In photodynamic therapy, spheroids lead to a better understanding of the own tumor response without interactions with vascular system. Finally, despite the quality of spheroid model, only the use of new technology for analysis of spheroid populations will help to increase their experimental use, particularly in preclinical oncology. PMID- 11033306 TI - Multicellular resistance: a paradigm for clinical resistance? AB - Research on resistance to cancer treatment was mainly focused for 20 years on multidrug resistance (MDR). No useful method of reversing MDR, suitable for clinical use, has yet emerged from this large quantity of work. The reason could be an inadequate evaluation of the target. When grown in spheroids, cancer cells exhibit a phenomenon known as 'multicellular resistance' (MCR). Tumours in patients seem to present the same characteristics. The mechanisms underlying MCR can be classified into two forms: contact resistance and resistance inherent in the spheroid structure. Mechanisms of MCR include: inhibition of apoptosis, high proportion of quiescent cells, modulation of protein expression (including topoisomerases and repair enzymes), potential permeability problems, presence of a hypoxic and necrotic centre and other possible mechanisms that remain to be discovered. A new therapeutic class of drugs is required to overcome MCR. Compounds, which are able to disrupt communication and binding between tumour cells and their microenvironment, seem to be able to circumvent MCR. Interesting results are obtained in vitro and in vivo in mice with specific antibodies or peptides recognised by cell binding proteins. Interestingly, these compounds also appear to be able to inhibit metastasis. Hyaluronidase has already been used with anticancer drugs in patients and was shown to increase drug potency. The explanation given is that it improves drug penetration into spheroids. We now hypothesise that hyaluronidase, in fact, decreases MCR and thus could be the first member of a new therapeutic class. PMID- 11033307 TI - Abstracts from the 7th Biennial Meeting of the International Society for Applied Cardiovascular Biology. March 9-11, 2000. Tucson, Arizona, USA. PMID- 11033308 TI - Preface (Festschrift david De wied) PMID- 11033309 TI - David and Goliath - the slingshot that started the neuropeptide revolution. AB - This review in honor of David de Wied summarizes the work done in my laboratory that first indicated that adrenocorticotropic hormone (ACTH) has a direct effect on the neuromuscular system. Cold stress or ACTH and its related peptides alpha melanocyte-stimulating hormone (alpha-MSH ) and beta-lipotropin improve the electromechanical characteristics of adrenalectomized and hypophysectomized rats. ACTH-(1-39) accelerates the return of motor and sensory function and improves the morphological characteristics of the motor endplate after peripheral nerve crush. The non-corticotropic fragments ACTH-(4-10), alpha-MSH, the ACTH-(4-9) analogue Organon 2766 (Org 2766) or the ACTH-(4-10) analogue Biomeasure 22015 (BIM 22015) improve electrophysiological and morphological parameters of the regenerating neuromuscular system. ACTH-(4-10) immunoreactivity, present in ventral horn motor neurons in low levels, is decreased ipsilaterally following ipsilateral nerve crush but increases both ipsilaterally and contralaterally if injured animals are treated with ACTH-(4-10) indicating a neuroprotective action. Similarly, Org 2766 appears to have a protective action in the brain following nigrostriatal lesions. In developmental studies, perinatal exposure to ACTH peptides improves the structure of the neuromuscular junction, accelerates the maturation of electromechanical properties and enhances nerve-muscle integration and nerve regeneration. Perinatal exposure to these peptides decreases adult male sexual behavior, a change correlated with increased serotinergic input within the medial preoptic area. Similar changes occur in female rats and appear to be long lasting. In tissue culture studies, both Org 2766 and BIM 22015 promote neurite outgrowth in the absence of nerve growth factor, indicating a neurotrophic role for these peptides. PMID- 11033310 TI - Melanocortins and the brain: from effects via receptors to drug targets. AB - The lack of specific receptors (and antagonists) has hampered the research on the neural mechanism of action of adrenocorticotropic hormone (ACTH)- and melanocyte stimulating hormone (MSH)-like peptides. Yet the original observations in the 1970s already pointed to cAMP as a possible mediator of ACTH/MSH effects in neurons. The cloning of melanocortin receptors since 1992, the identification of at least two subtypes (melanocortin MC(3) and MC(4) receptors) that are present in neural tissue and the development of selective and potent agonists as well as antagonists have markedly furthered the position of melanocortins as important neuropeptides. In this paper we discuss the role of especially the receptor subtype melanocortin MC(4) in various behaviors including grooming behavior and feeding behavior and consider new insights in the interaction between the opioid and the melanocortin system at the level of the spinal cord (i.e. pain perception). Finally, based on new data obtained in molecular pharmacological studies on brain melanocortin receptors, we suggest a general concept for selective receptor-ligand interaction: ligand residues outside the peptide core sequence may direct the conformation of the residues in the ligand core-sequence that interact directly with the receptor-binding pocket and thereby determine selectivity. PMID- 11033311 TI - Role of melanocortins in the central control of feeding. AB - The injection of a melanocortin peptide or of melanocortin peptide analogues into the cerebrospinal fluid or into the ventromedial hypothalamus in nanomolar or subnanomolar doses induces a long-lasting inhibition of food intake. The effect keeps significant for up to 9 h and has been observed in all animal species so far tested, the most susceptible being the rabbit. The anorectic effect of these peptides is a primary one, not secondary to the shift towards other components of the complex melanocortin-induced behavioral syndrome, in particular grooming. The site of action is in the brain, and the effect is not adrenal-mediated because it is fully exhibited also by adrenalectomized animals. It is a very strong effect, because the degree of feeding inhibition is not reduced in conditions of hunger, either induced by 24 h starvation, or by insulin-induced hypoglycemia, or by stimulation of gamma-aminobutyric acid (GABA), noradrenergic or opioid systems. The microstructural analysis of feeding behavior suggests that melanocortins act as satiety-inducing agents, because they do not significantly modify the latencies to start eating, but shorten the latencies to stop eating. The mechanism of action involves the activation of melanocortin MC(4) receptors, because selective melanocortin MC(4) receptor antagonists inhibit the anorectic effect of melanocortins, while inducing per se a strong stimulation of food intake and a significant increase in body weight. Melanocortins seem to play an important role in stress-induced anorexia, because such condition, in rats, is significantly attenuated by the blockage of melanocortin MC(4) receptors; such a role is not secondary to an increased release of corticotropin-releasing factor (CRF), because, on the other hand, the CRF-induced anorexia is not affected at all by the blockage of melanocortin MC(4) receptors. The physiological meaning of the feeding inhibitory effect of melanocortins, and, by consequence, the physiological role of melanocortins in the complex machinery responsible for body weight homeostasis, is testified by the hyperphagia/obesity syndromes caused by mutations in the pro-opiomelanocortin (POMC) gene, or in the melanocortin MC(4) receptor gene, or in the agouti locus. Finally, recent evidences suggest that melanocortins could be involved in mediating the effects of leptin, and in controlling the expression of neuropeptide Y (NPY). PMID- 11033312 TI - Postnatal treatment with ACTH-(4-9) analog ORG 2766 attenuates N-methyl-D aspartate-induced excitotoxicity in rat nucleus basalis in adulthood. AB - It has been reported that the ACTH-(4-9) analog H-Met(O(2))-Glu-His-Phe-D-Lys-Phe OH (ORG 2766) administered in adulthood has trophic effects on neuronal tissue and when given postnatally, it can induce long-lasting changes in brain development. In the present study, we investigated whether early postnatal treatment with ORG 2766 affects adult neuronal vulnerability, i.e. the sensitivity of cholinergic neurons against excitotoxic damage. Wistar rat pups received injections of ORG 2766 or saline on postnatal days 1, 3 and 5 and were then left undisturbed until adulthood. At the age of 6 months, the animals were subjected to unilateral lesion of magnocellular basal nucleus by infusion of high dose of N-methyl-D-aspartate (NMDA). The effects of the excitotoxic insult were studied 28 hours and 12 days after the lesion by measuring both the acute cholinergic and glial responses, and the final outcome of the degeneration process. Twenty eight hours after NMDA infusion, postnatally ACTH-(4-9)-treated animals showed stronger suppression of choline-acetyltransferase immunoreactivity and increased reaction of glial fibrillary acidic protein -immunopositive astrocytes in the lesioned nucleus compared to control animals. However, 12 days post-surgery, the NMDA-induced loss of cholinergic neurons, as well as the decrease of their acetylcholinesterase -positive fibre projections in the cortex, were less in ACTH-(4-9) animals. Our data indicate that the early developmental effects of ACTH-(4-9) influence intrinsic neuroprotective mechanisms and reactivity of neuronal and glial cells, thereby resulting in a facilitated rescuing mechanism following excitotoxic injury. PMID- 11033313 TI - Manipulating neuropeptidergic pathways in humans: a novel approach to neuropharmacology? AB - Given the tremendous number of neuropeptides, which are synthesized in the central nervous system, the brain can be viewed as one of the most prominent endocrine organs. Elucidation of the functions of these peptides is hampered by the facts that after intravenous administration access to brain receptors is prevented or impaired by the blood-brain barrier. Here, we provide evidence that intranasal administration can be a way to circumvent the blood-brain barrier. Selected experiments will be reported indicating that peptides after intranasal administration in humans can specifically alter a great variety of brain functions. For vasopressin, we demonstrated improving effects of long-term intranasal treatment on sleep in elderly people. Insulin showed improving effects of short-term memory functions. For adrenocorticotropin/melanocyte stimulating hormone, ACTH/MSH-(4-10), a twofold action was isolated: The melanocortin fragment diminished selective attention and, with subchronic administration, reduced body fat. These results could provide the basis for developing a new, specific, and "soft" neuropharmacology. PMID- 11033314 TI - Genetic pathways in the developmental specification of hypothalamic neuropeptide and midbrain catecholamine systems. AB - The neuropeptide concept concerns the diverse and broad physiological functions of neuropeptides in behavioral adaptation. Neuropeptides like vasopressin and corticotropin-releasing hormone can coordinate multiple brain functions due to the anatomical organization of the neurons producing them. The cell bodies are focally positioned in the hypothalamus and send long-reaching efferents to limbic and brainstem areas. Likewise, midbrain dopamine systems coordinate emotional behaviors and movement control by specific connectivity of neurons in the midbrain to limbic and striatal centers, respectively. The fundament of the functions of these signalling molecules is laid out during development when transmitter identity and connectivity are specified. This is a highly controlled process involving multiple transcription factors and growth factors acting together in genetic pathways. Here, the genetic pathways enrolling in developing vasopressin, corticotropin-releasing hormone, and midbrain dopamine neurons are discussed. PMID- 11033315 TI - Neuromodulation of memory in the hippocampus by vasopressin. AB - The involvement of [Arg(8)]vasopressin in memory processes was analyzed in the hippocampal structure, since we have reported that this is one of the main central target structures of the vasopressin-enhancing effect on memory. This structure is functionally differentiated along its dorsoventral axis, and the expression of the vasopressinergic system is dependent upon whether the dorsal or ventral part of the hippocampus is involved. For this reason, the effect of vasopressin injected into hippocampus was evaluated on the basis of the site of injection. We have shown, using a Go-No Go visual discrimination task with mice that both parts of the hippocampus are involved in the effect of endogenous or exogenous vasopressin, but with higher sensitivity for the ventral part. Based on the expression of Fos protein following intracerebroventricular injection of vasopressin in unconditioned or conditioned mice, we confirmed the greater involvement of the ventral hippocampus in the enhancing effect of vasopressin on memory processes. The effect of the peptide seems specific, since only a few of the hippocampal cells that expressed Fos protein in the unconditioned mice did so in the conditioned mice (cells in the dentate gyrus and the CA3 hippocampal field). Moreover, we have shown that in the ventral hippocampus, vasopressin generates different behavioral effects whether treatment is performed at the beginning or in the middle of the learning process, suggesting that the mnemonic context is an important factor for understanding the effect of vasopressin on memory in the ventral hippocampus. PMID- 11033316 TI - Spatial, cellular and temporal basis of vasopressin potentiation of norepinephrine-induced cAMP formation. AB - This study investigated the spatial distribution of vasopressin V(1) and beta(1) adrenoceptors within hippocampal subfields and lamina in an attempt to localize the site(s) of interaction between these two receptor systems. In addition, the cell types, neuronal and glial, in which the vasopressin-induced neuromodulation occurs, were identified. Lastly, the temporal constraints of the potentiation induced by vasopressin were investigated. Results of these analyses demonstrated multiple sites within the hippocampus where the interaction between vasopressin and norephinephrine could occur. Moreover, vasopressin-induced potentiation of adrenergic stimulated cyclase occurred in both hippocampal neurons and glia whereas it did not occur in undifferentiated neurons. Analysis of the temporal constraints of vasopressin-induced potentiation revealed that pre-activation of the vasopressin V(1) receptor for 1 min yielded greater potentiation than simultaneous exposure to vasopressin and norepinephrine. These data provide insights into the spatial and temporal characteristics for the interaction between the vasopressin receptor and adrenoceptor systems and provide a cellular and biochemical rationale for the behavioral findings of Kovacs and De Wied. PMID- 11033317 TI - Endogenous opioids and reward. AB - The discovery of endogenous opioids has markedly influenced the research on the biology of addiction and reward brain processes. Evidence has been presented that these brain substances modulate brain stimulation reward, self-administration of different drugs of abuse, sexual behaviour and social behaviour. There appears to be two different domains in which endogenous opioids, present in separate and distinct brain regions, are involved. One is related to the modulation of incentive motivational processes and the other to the performance of certain behaviours. It is concluded that endogenous opioids may play a role in the vulnerability to certain diseases, such as addiction and autism, but also when the disease is present, such as alcoholism. PMID- 11033318 TI - The role of atrial natriuretic peptide in alcohol withdrawal: a peripheral indicator and central modulator? AB - Changes in fluid and electrolyte homeostasis may accompany and are likely to modify the clinical symptoms of alcohol-withdrawal reactions. It was of obvious theoretical and practical interest therefore to investigate the changes in the secretion of hormones, which regulate the fluid and electrolyte homeostasis (atrial natriuretic peptide, aldosterone and plasma renin activity) during alcohol withdrawal in chronic alcoholic patients. In a phase of severe withdrawal, there were increased plasma renin activity and aldosterone levels observed. In a phase of partial recovery, on the other hand, the elevated plasma renin activity and aldosterone levels were back to the normal range. In 60% of the patients, delirium tremens was gradually developing during the observation period. In these patients, an elevated level of atrial natriuretic peptide was observed at the time of hospital admission, i.e. days before the actual onset of delirium tremens. It is concluded that the disturbed volume homeostasis and the consequently altered plasma atrial natriuretic peptide secretion might be associated with, and therefore used as an indicator of the onset of delirium tremens. To study the role of central nervous atrial natriuretic peptide, mice were rendered tolerant to and dependent on alcohol with an alcohol-liquid diet for 14 days. Five hours after withdrawal from alcohol, withdrawal hyperexcitability symptoms were analyzed. Intracerebroventricular (i.c.v.) injection of atrial natriuretic peptide attenuated, whereas that of an antiserum against atrial natriuretic peptide intensified the severity of handling-induced convulsions. N-methyl-D-aspartate induced behavioral seizures in a dose-dependent manner, whose effect was more intensive during the alcohol-withdrawal period than in alcohol-naive animals. I.c.v. injections of atrial natriuretic peptide dose dependently inhibited, whereas that of antiserum against atrial natriuretic peptide potentiated the seizure-inducing effect of N-methyl-D-aspartate in alcohol-dependent mice. Although tentatively, it is concluded that peripheral secretion of atrial natriuretic peptide may be an indicator, whereas central nervous atrial natriuretic peptide a neuropeptide modulator of alcohol-withdrawal symptomatology. PMID- 11033319 TI - Excitatory non-adrenergic-non-cholinergic neuropeptides: key players in asthma. AB - Professor David de Wied first introduced the term 'neuropeptides' at the end of 1971. Later peptide hormones and their fragments, endogenous opioid (morphine like) peptides and a large number of other biogenic peptides became classified as neuropeptides. All of these peptides are united by a number of common features including their origin (nervous system and peptide-secreting cells found in various organs such as skin, gut, lungs), biosynthesis, secretion, metabolism, and enormous effectiveness. Neuropeptides are biologically active at extremely low concentrations. The past decade, neuropeptide research has revealed that neuropeptides also participate strongly in immune reactions. The neuro-immune concept has opened up a whole new research area. In the last 20 years, significant advances have been made in investigations of the interaction between immune and nervous systems in chronic inflammatory diseases such as asthma. The goal of this review is to bring together the functional relevance of excitatory non-adrenergic-non-cholinergic (NANC) nerves and the interaction with the immune system in asthma. PMID- 11033320 TI - The "low-dose" concept and the paradoxical effects of prolactin on grooming and sexual behavior. AB - The effects of prolactin on animal behavior include the stimulation of novelty induced grooming in rats. This effect has been demonstrated in hyperprolactinaemic animals bearing pituitary homografts under the kidney capsule or after intracerebroventricular (i.c.v.) administration of prolactin. Since plasma prolactin levels in hyperprolactinaemic rats are similar to those of animals injected with low doses of rat prolactin, we studied the effects of this hormone injected subcutaneously (s.c.) in a dose range of 5-50 microg/kg. Novelty induced grooming was enhanced only in rats injected with 5 or 10 microg/kg rat prolactin, whereas no effect was observed after the s.c. injection of the higher dose. The sexual behavior of male rats is also affected by prolactin. Male rats with normal mating activity showed enhanced sexual behavior when injected s.c. with rat prolactin (5, 10 or 50 microg/kg). In animals with poor sexual performance or in impotent rats, prolactin (5 or 10 microg/kg, but not 50 microg/kg) restored the full pattern of sexual behavior. An increased lordosis quotient was also observed in ovariectomized rats treated with prolactin 5 or 10 microg/kg. These results suggest that, besides the duration of hyperprolactinaemia, the effective level of plasma prolactin is important for the expression of the behavioral effects of this hormone. PMID- 11033321 TI - The glucocorticoid hormone: from pedestal to dust and back. AB - A potential injury to the hippocampus has been postulated by the "glucocorticoid cascade hypothesis" as deriving from the life-long exposure to the stress glucocorticoid hormone. This hypothesis has been extensively resorted to in the search of a physio-pathological basis of the cognitive and behavioural impairments of old age, as well as for assigning to the hormone a not-irrelevant pathogenic role in brain degenerative diseases. Here I discuss the experimental evidences that have credited to stress a killing-licence, and pose, on the contrary, that the modest degrees of hypercortisolemia present in the above conditions could be interpreted as a beneficial occurrence. PMID- 11033322 TI - Influence of psychological variables on the activity of the hypothalamic pituitary-adrenal axis. AB - Psychobiology is the discipline that attempts to integrate the impact of environmental and psychological variables on biological systems. This paper focuses on the psychobiology of the hypothalamic-pituitary-adrenal (HPA) axis and illustrates several processes that influence the response of the HPA axis. The interaction of the developing rodent or primate with their primary care giver has permanent long-term effects on the HPA axis. Manipulations that alter maternal behavior during critical periods of development permanently modify the HPA axis. The HPA axis can be programmed to be hypo-responsive or hyper-responsive as a function of time and length of maternal separation. In the adult organism, the HPA response to stress is highly dependent on specific psychological factors such as control, predictability, and feedback. In primates, social variables have been shown to diminish or exacerbate the HPA stress response. During the post-natal period of development, the mother appears to actively inhibit the pups' HPA axis. Different aspects of maternal behavior regulate different components of the HPA system. PMID- 11033323 TI - Stress-induced expression of co-localized neuropeptides in hypothalamic and amygdaloid neurons. AB - This short review summarizes the effect of various stressful stimuli on the expression of neuropeptides which co-localize in corticotrophin releasing hormone (CRH)-synthesizing neurons in the hypothalamic paraventricular nucleus, as well as in oxytocin and vasopressin neurons in the supraoptic nucleus. Stress-induced changes failed to act on CRH neurons in the central amygdaloid nucleus but formalin-evoked pain enhanced galanin mRNA expression in the medial subdivision of this nucleus. Changes in the expression of enkephalin, galanin, dynorphin and cholecystokinin mRNA in response to restraint and formalin-induced pain are documented in hypothalamic and amygdaloid nuclei by in situ hybridization histochemical technique. PMID- 11033324 TI - On the development of psychoneuroimmunology. AB - Psychoneuroimmunology, the study of interactions among behavioral, neural and endocrine, and immune processes, coalesced as an interdisciplinary field of study in the late 1970s. Some of the early research that was critical in establishing neuroanatomical, neurochemical and neuroendocrine pathways and functional relationships between the brain and the immune system is outlined here. These and subsequent studies have led to the general acknowledgment that the nervous and immune systems are components of an integrated system of adaptive processes, and that immunoregulatory processes can no longer be studied as the independent activity of an autonomous immune system. This paradigm shift in the study of immunoregulatory processes and the elaboration of the mechanisms underlying behaviorally induced alterations of immune function promise a better understanding and a new appreciation of the multi-determined etiology of pathophysiological states. PMID- 11033325 TI - Non-linear effects in the retention of an avoidance task induced by anabolic steroids. AB - The results of the study reported in Brain Research in 1995 by Isaacson et al. [Isaacson, R.L., Varner, J.A., Baars, J.-M., de Wied, D., 1995. The effects of pregnenolone sulfate and ethylestrenol on retention of a passive avoidance task. Brain Res. 689, 79-84] have been re-examined with special emphasis placed on the distributions of latencies found in the passive avoidance task using rats. This study used two retention tests, one 24 h after training the other at 48 h after training. In the first experiment in that study a range of doses of two anabolic steroids, pregnenolone sulfate and ethylestrenol, were given s.c. just after the footshock training trial. In experiment 2 a similar range of doses of both steroids was given to the rats 1 h before the first retention test. Placing emphasis on the distributions rather than measures of central tendencies revealed that, in contrast to the vehicle treated animals, the anabolic steroid treated animals exhibited bimodal distributions of response latencies. These differences between control and hormone treated animals were observed in both experiments. The new information was interpreted in terms of non-linear dynamics including some aspects of Chaos theory. PMID- 11033326 TI - Stress in the brain. AB - Part I (first section) reports about research in the period 1964-1976, when the seminal observations were made on which today's concept of corticosteroid action on the brain is based. These key observations concern the discovery of nuclear corticosterone receptors in the limbic brain that mediate control over neuronal circuits underlying hypothalamic-pituitary-adrenal activity and behavioural adaptation. Part II (second section) covers the period of 1977-1989. It is about some aspects of the neuropeptide concept, the implementation of micro neurochemistry using the "Palkovits punch", and the application of in vitro autoradiography. Vasopressin and oxytocin receptors were identified and their implication in behaviour was examined using the song control of the canary bird as a model system. Two distinct nuclear receptor types for corticosteroids were identified: mineralocorticoid receptors (MR) and glucocorticoid receptors (GR) which mediate in a coordinate manner the steroid control of hypothalamus pituitary-adrenal activity and behaviour. Part III (third section) is from 1990 up to 2000. Focus is on the balance of MR- and GR-mediated actions in control of homeostasis as a determinant of health and disease. MR operates in pro-active mode to prevent homeostatic disturbance, while additional GR activation promotes in reactive fashion recovery after stress. An imbalance in MR and GR underlies behavioural deficits and neuroendocrine disturbances increasing vulnerability for stress-related brain disorders. The complete hippocampal genome is screened for corticosteroid responsive genes, which are potential targets for drugs promoting restorative capacity still present in the diseased brain. PMID- 11033327 TI - The role of CRF receptor subtypes in stress-induced behavioural responses. AB - The actions of corticotropin-releasing factor (CRF) and CRF-related peptides in the brain and periphery are mediated through multiple receptors. Two CRF receptor subtypes that differ markedly in their pharmacological profiles and anatomical distribution have been identified and characterized. Important advances have been made in understanding CRF and its actions through the development of specific CRF receptor antagonists, application of antisense oligonucleotides, and the production of transgenic mice lacking functional CRF(1) receptors. This chapter describes recent findings with respect to CRF-related peptides and CRF receptors and their role in stress-induced behaviours. PMID- 11033328 TI - Modulatory actions of steroid hormones and neuropeptides on electrical activity in brain. AB - Electrophysiological studies over the past decades have shown that many compounds in addition to 'classical' neurotransmitters affect electrical activity in the brain. These compounds include neuropeptides synthesized in brain as well as compounds which are released from peripheral sources and subsequently enter the brain compartment, such as corticosteroid hormones from the adrenal gland. In the present review, this principle is illustrated by describing the effects of two substances, i.e. vasopressin and corticosterone. Neuropeptides and corticosteroid hormones add at least two essential aspects to information processing in the brain. First, they both act conditional, i.e. they modulate the actions of 'classical' neurotransmitters, rather than changing basal neuronal activity by themselves. Second, the time-frame in which modulation of electrical properties takes place differs from that generally seen with 'classical' neurotransmitters. Neuropeptides modulate electrical activity over a period of minutes, while effects of corticosteroid hormones usually become apparent after at least an hour but then last for hours. In this way, neuropeptides and steroid hormones expand the repertoire of responses through which the brain reacts to environmental challenges. PMID- 11033329 TI - Long-lasting stress sensitisation. AB - Stressful experiences in humans can result in a spectrum of long-term changes in behavioural, autonomic and hormonal responsivity. An extreme form of such alterations is found in patients with post-traumatic stress disorder (PTSD). A number of animal models has been developed in which intense stressful experiences (shocks, social confrontations) result in longterm altered responsivity of behavioural, autonomic and hormonal responses to aversive challenges which mimic many of the changes seen in PTSD. These models of stress-induced sensitisation are beginning to generate a better understanding of the vulnerability factors, time-course and underlying neuronal substrates of the long-term disturbances experienced by humans as a result of stressful life events. PMID- 11033330 TI - Role of corticotropin-releasing factor, vasopressin and the autonomic nervous system in learning and memory. AB - Learning and memory are essential requirements for every living organism in order to cope with environmental demands, which enables it to adapt to changes in the conditions of life. Research on the effects of hormones on memory has focused on hormones such as adrenocorticotropic hormone (ACTH), glucocorticoids, vasopressin, oxytocin, epinephrine, corticotropin-releasing factor (CRF) that are released into the blood and brain following arousing or stressful experiences. Most of the information have been derived from studies on conditioned behavior, in particular, avoidance behavior in rats. In these tasks, an aversive situation was used as a stimulus for learning. Aversive stimuli are associated with the release of stress hormones and neuropeptides. Many factors play a role in different aspects of learning and memory processes. Neuropeptides not only affect attention, motivation, concentration and arousal or vigilance, but also anxiety and fear. In this way, they participate in learning and memory processes. Furthermore, neuropeptides such as CRF and vasopressin modulate the release of stress hormones such as epinephrine. In turn, systemic catecholamines enhance memory consolidation. CRF and vasopressin are colocalized in neurons from the nucleus paraventricularis, which project to nuclei in the brainstem involved in autonomic regulation. The objective of this paper is to discuss the role of CRF, vasopressin, and the autonomic nervous system (ANS) in learning and memory processes. Both CRF and vasopressin have effects in the same direction on behavior, learning and memory processes and stress responses (release of catecholamines and ACTH). These neuropeptides may act synergistically or in a concerted action aimed to learn to adapt to environmental demands. PMID- 11033331 TI - The brain mineralocorticoid receptor: greedy for ligand, mysterious in function. AB - Glucocorticoids exert their regulatory effects on the hypothalamic-pituitary adrenocortical axis via two types of corticosteroid receptors: the glucocorticoid receptor and the mineralocorticoid receptor. Whereas the glucocorticoid receptor has a broad distribution in the brain, highest levels of mineralocorticoid receptor are found in the hippocampus. Based on the differential occupancy profile by endogenous glucocorticoids, glucocorticoid receptors are thought to mediate negative feedback signals of elevated glucocorticoid levels, whereas mineralocorticoid receptors control the inhibitory tone of the hippocampus on hypothalamic-pituitary-adrenocortical axis activity. Dysfunction of mineralocorticoid receptors and glucocorticoid receptors are thought to be implicated in stress-related psychiatric diseases such as major depression. Because of its intriguing features, we focus in this review on the mineralocorticoid receptor and provide data which reveal novel aspects of the pharmacology and physiology of mineralocorticoid receptors. Newly obtained results are presented, which help to solve the paradox of why dexamethasone binds with high affinity to mineralocorticoid receptors in vitro, yet binds poorly in vivo. Until recently, mineralocorticoid receptor protein and mRNA levels could only be routinely studied with in vitro cytosol binding assays, in vitro and in vivo receptor autoradiography, Northern blot analysis, and in situ hybridization. These methods are unfortunately hampered by several flaws, such as the necessity of adrenalectomy, no or poor neuroanatomical resolution, the fact that mRNA does not provide the same information as protein, or combinations of these factors. We present immunohistochemical data on mineralocorticoid receptors in the brain obtained by using commercially available antibodies, which alleviate many of these shortcomings. Furthermore, an in vivo microdialysis method is presented which allows the assessment of free corticosterone levels in the brain, which is critical for the study of the pharmacological basis of mineralocorticoid receptor (and glucocorticoid receptor) function. Finally, a novel aspect of the regulation of mineralocorticoid receptors is described which provides evidence that this receptor system is dynamically regulated. In conjunction with previously reported effects of antidepressants, these results have initiated a new concept on the cause of the hypothalamic-pituitary-adrenocortical axis disturbances often seen in stress-related psychiatric disorders such as major depression. PMID- 11033332 TI - Sexual dimorphism in immune function: the role of prenatal exposure to androgens and estrogens. AB - Perinatal exposure to androgens permanently transforms certain tissues, e.g., the brain, the genitalia, etc. This process involves both masculinization and defeminization. Immune function also is transformed by early steroid exposure; however, it is not yet known whether the response capabilities of the immunocytes themselves are directly modified or whether they are responding to signals from other masculinized tissues, e.g., the brain. Most evidence points to a direct effect since androgen and estrogen receptors are present in developing immunocytes. Both androgens and estrogens have a role in regulating adult immunity including Th1/Th2 balance. Adult susceptibility to autoimmune and other diseases is also related to steroid exposure. How immune cells respond to gonadal steroids in adulthood may depend on the pattern of androgenic and estrogenic stimulation during early development. PMID- 11033333 TI - Stages of avoidance strategy formation in gerbils are correlated with dopaminergic transmission activity. AB - This detailed analysis of behavior is aimed at the differentiation of the components of information processing during associative conditioning. In gerbils, the influences of various acquired non-avoidance strategies as pre-experience were studied during the learning of a standard avoidance task in the same shuttle box. Identical cue stimuli, frequency-modulated tones as conditioned stimuli and electric footshocks as unconditioned stimuli, were used in various behavioral tasks. In addition to common parameters such as avoidance performance and reaction times, behavioral events such as the attention response and the orienting response were quantified. Thereby, components of shuttle-box learning such as signal detection and signal evaluation were found to be affected by pre experience-dependent dynamics. Using a microdialysis technique during avoidance learning in the shuttle-box, we found that only strategy formation was correlated with high dopamine levels in medial prefrontal cortex. The increase in dopamine in medial prefrontal cortex may be an indicator of the involvement of working memory principles in signal evaluation stages of conditioning. PMID- 11033334 TI - Platelets as a peripheral district where to study pathogenetic mechanisms of alzheimer disease: the case of amyloid precursor protein. AB - Alzheimer disease is a progressive neurodegenerative disease, characterised by a progressive cognitive and memory decline. From a neuropathological point of view, Alzheimer disease is defined by the presence of characteristic lesions, i.e. mature senile plaques, neurofibrillary tangles (NFTs) and amyloid angiopathy. In particular, accumulation of the amyloid beta-peptide in the brain parenchyma and vasculature is an invariant event in the pathogenesis of both sporadic and familial Alzheimer cases. Amyloid beta-peptide originates from a larger precursor, the amyloid precursor protein (APP) ubiquitously expressed. Among the different peripheral cells expressing APP forms, platelets are particularly interesting since they show concentrations of its isoforms equivalent to those found in brain. Moreover, a number of laboratories independently described alterations in APP metabolism/concentration in platelets of Alzheimer patients when compared to control subjects matched for demographic characteristics. These observations defined the frame of our work aimed to investigate if a correlation between levels of platelet APP forms and Alzheimer disease could be detected. We have reported that patients affected by Alzheimer disease show a differential level of platelet APP forms. This observation has several implications: APP processing abnormalities, believed to be a very early change in Alzheimer disease in neuronal compartment, do occur in extraneuronal tissues, such as platelets, thus, suggesting that Alzheimer disease is a systemic disorder; further, our data strongly indicate that a differential level of platelet APP isoforms can be considered a potential peripheral marker of Alzheimer disease allowing for discrimination between Alzheimer and other types of dementia. PMID- 11033335 TI - Motor neuron disease in vitro: the use of cultured motor neurons to study amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease in which motor neurons in the nervous system die. The cause is unknown, and no effective treatment exists. Mutations in the gene for superoxide dismutase found in a subpopulation have led to an animal model, but research with these mice has not produced complete insight into the disease mechanism. Studies with isolated motor neurons may produce important information. This review discusses approaches to culture motor neurons - single cells, cocultured with other cells, and in intact preparations, such as the spinal or cortical slice. Motor neurons in monoculture are suitable for acute but not for chronic studies, whereas cocultures and slices survive up to months and are used for chronic studies. Results with toxic substances believed to play a role in the disease, such as oxidants and glutamate, and of studies where the energy status of the cells is manipulated, are presented. PMID- 11033336 TI - Bone marrow: a possible alternative source of cells in the adult nervous system. AB - There is increasing evidence that stem cell populations can undergo a transition between mesodermal and neural ectodermal cell fates. Bone marrow-derived cells have been shown to be extremely versatile: they can become brain and liver cells and muscle, while other mesodermal derived cells have been shown to migrate into the brain and differentiate into neurons. Moreover, under the appropriate conditions, neural stem cells can differentiate into hematopoietic and muscle cell fates. It is now well established that newly differentiated cell types are continuously generated from immature stem cells throughout development and in adult mammals, including humans. This review addresses the contribution that bone marrow-derived stem cells may play during neurogenesis. We transplanted male bone marrow into female recipients to track and characterize the Y chromosome containing cells in the CNS (central nervous system) of mice. PMID- 11033337 TI - The dopamine D(4) receptor: one decade of research. AB - Dopamine is an important neurotransmitter involved in motor control, endocrine function, reward, cognition and emotion. Dopamine receptors belong to the superfamily of G protein-coupled receptors and play a crucial role in mediating the diverse effects of dopamine in the central nervous system (CNS). The dopaminergic system is implicated in disorders such as Parkinson's disease and addiction, and is the major target for antipsychotic medication in the treatment of schizophrenia. Molecular cloning studies a decade ago revealed the existence of five different dopamine receptor subtypes in mammalian species. While the presence of the abundantly expressed dopamine D(1) and D(2) receptors was predicted from biochemical and pharmacological work, the cloning of the less abundant dopamine D(3), D(4) and D(5) receptors was not anticipated. The identification of these novel dopamine receptor family members posed a challenge with respect to determining their precise physiological roles and identifying their potential as therapeutic targets for dopamine-related disorders. This review is focused on the accomplishments of one decade of research on the dopamine D(4) receptor. New insights into the biochemistry of the dopamine D(4) receptor include the discovery that this G protein-coupled receptor can directly interact with SH3 domains. At the physiological level, converging evidence from transgenic mouse work and human genetic studies suggests that this receptor has a role in exploratory behavior and as a genetic susceptibility factor for attention deficit hyperactivity disorder. PMID- 11033338 TI - Structure and function of the dopamine transporter. AB - The dopamine transporter mediates uptake of dopamine into neurons and is a major target for various pharmacologically active drugs and environmental toxins. Since its cloning, much information has been obtained regarding its structure and function. Binding domains for dopamine and various blocking drugs including cocaine are likely formed by interactions with multiple amino acid residues, some of which are separate in the primary structure but lie close together in the still unknown tertiary structure. Chimera and site-directed mutagenesis studies suggest the involvement of both overlapping and separate domains in the interaction with substrates and blockers, whereas recent findings with sulfhydryl reagents selectively targeting cysteine residues support a role for conformational changes in the binding of blockers such as cocaine. The dopamine transporter can also operate in reverse, i.e. in an efflux mode, and recent mutagenesis experiments show different structural requirements for inward and outward transport. Strong evidence for dopamine transporter domains selectively influencing binding of dopamine or cocaine analogs has not yet emerged, although the development of a cocaine antagonist at the level of the transporter remains a possibility. PMID- 11033339 TI - Psychoneurendocrinology: a science of the past or a new pathway for the future? AB - Psychoneuroendocrinology is a branch of neuroscience that developed in the beginning of the last century, which investigates the possibility of a cause effect link between endocrinopathies and mental disorders - with these studies ending in negative results. Psychoneuroendocrinology was then used as a methodological approach for the investigation of neurotransmitter function, on the basis of the observation that neurotransmitters regulate neurohormone and peripheral hormone secretions. Data were obtained for hypothalamic noradrenergic, serotoninergic, dopaminergic, gabaergic and acetylcholinergic functions, which could not be automatically extended to higher brain centers whose impairments might be etiopathogenetically involved in the development of mental disorders. Future studies should focus on new methodological approaches to brain biochemistry, on investigation of genetic, molecular biology, brain imaging, psychoneuroimmunoendocrinology, neuropeptide and neurosteroid secretion in relation to brain endocrine function in mental diseases. PMID- 11033340 TI - Antidepressants: past, present and future. AB - Since the discovery of first antidepressants in mid-1950's, the field has been intensively studied. Several new classes of compounds emerged and several hypotheses on the mechanism of their action were proposed. The novel antidepressants are either selective and reversible monoamine oxidase inhibitors, (e.g., moclobemide), or selective serotonin reuptake inhibitors (e.g., citalopram or paroxetine), or serotonin and noradrenaline reuptake inhibitors (e.g. , venlafaxine). Recently neuropeptides (e.g., thyrotropin-releasing hormone,TRH) or antagonists of neuropeptide receptors (e.g., tachykinin NK(1) receptor) undergo clinical tests. Several hypotheses proposed the predominant involvement of one or few neurotransmitter receptors in the mechanism of antidepressant action, but it is now assumed that several distinct receptor mechanisms' trigger different but converging intracellular signal cascades that activate transcription factors, which, in turn, promote the expression of genes encoding for proteins, that play a crucial role in restoring of neuronal functions involved in mood regulation. PMID- 11033341 TI - The role of dopamine in the mechanism of action of antidepressant drugs. AB - The present paper reviews evidence on the effect of antidepressant treatments on dopamine transmission. Chronic treatment with antidepressant drugs potentiates the behavioural stimulant responses elicited by the stimulation of dopamine receptors, including reward-related behaviours. Moreover, antidepressants affect dopamine release in several brain areas. The reviewed literature is discussed in terms of the possible mechanisms underlying antidepressant-induced supersensitivity to dopamine-mediated behavioural responses, and of the possible implications for the therapeutic effect of these drugs. It is concluded that the potentiation of dopaminergic neurotransmission induced by chronic antidepressant treatments might contribute to their therapeutic effect. PMID- 11033342 TI - Stress in schizophrenia: an integrative view. AB - Stress and the development of a (schizophrenic) psychosis are inextricably related. The process by which stress actually affects psychosis is far less clear. The hypothalamic-pituitary-adrenal system, and in particular the release of corticosteroids, has been attributed an essential role. However, schizophrenia is a disorder in which many functions are distorted. Dysfunctions can be found in behavior, cognition, coping, physiology, pituitary-adrenal and immune functioning. In this short paper, these functions are discussed as to how they contribute to the way stress is appraised and processed. Schizophrenic patients are impaired in their biological response to stress by showing a blunted cortisol response to psychosocial stress. It is hypothesized that this reflects rather cognitive dysfunction, based on biological dysfunctions in those brain structures that are responsible for these processes, i.e. the prefrontal cortex and the limbic system. Considering the blunted cortisol response as a maladaptive stress response, its consequences are commented on with an emphasis on the immune system. Finally, the role of neuroleptics, and in particular the atypical ones, is discussed for their beneficial effect, beyond their fear-and anxiety-reducing properties, in restoring some of the cognitive dysfunctions schizophrenic patients display. By doing so, they may improve perception of the environment, enhance adjustment and thus a proper stress response. Integration of these processes in stress research described, may provide new vistas of the stress concept in schizophrenia. PMID- 11033343 TI - Dysregulation of the neural cell adhesion molecule and neuropsychiatric disorders. AB - Cell adhesion molecule proteins play a diverse role in neural development, signal transduction, structural linkages to extracellular and intracellular proteins, synaptic stabilization, neurogenesis, and learning. Three basic mRNA isoforms and potent posttranslational modifications differentially regulate these neurobiological properties of the neural cell adhesion molecule (N-CAM). Abnormal concentrations of N-CAM 105-115 kDa (cN-CAM), N-CAM variable alternative spliced exon (VASE), and N-CAM secreted exon (SEC) are related to schizophrenia and bipolar neuropsychiatric disorders. These N-CAM isoforms provide potential mechanisms for expression of multiple neurobiological alterations between controls and individuals with schizophrenia or bipolar illness. Multiple processes can trigger the dysregulation of N-CAM isoforms. Differences in neuropil volume, neuronal diameter, gray matter thickness, and ventricular size can be related to N-CAM neurobiological properties in neuropsychiatric disorders. Potential test of the N-CAM dysregulation hypothesis of neuropsychiatric disorder is whether ongoing dysregulation of N-CAM would cause cognitive impairments, increased lateral ventricle volume, and decreased hippocampal volume observed in schizophrenia and to a lesser extent in bipolar disorder. An indirect test of this theory conducted in animal experiments lend support to this N-CAM hypothesis. N-CAM dysregulation is consistent with a synaptic abnormality that could underlie the disconnection between brain regions consistent with neuroimaging reports. Synapse stability and plasticity may be part of the molecular neuropathology of these disorders. PMID- 11033344 TI - Noradrenaline systems in the hypothalamus, amygdala and locus coeruleus are involved in the provocation of anxiety: basic studies. AB - A variety of stressful events, including emotional stress, cause a marked increase in noradrenaline release in several brain regions, and especially in the hypothalamus, amygdala and locus coeruleus, in the rat brain. These findings suggest that an increased noradrenaline release could be closely related to the provocation of negative emotions such as anxiety and/or fear. In order to confirm this hypothesis, we carried out several studies. Diazepam, a typical benzodiazepine anxiolytic, significantly attenuated not only the immobilization stress-induced increase in noradrenaline release in the three rat brain regions but also the emotional changes of these animals, and these effects were antagonized by flumazenil, a benzodiazepine antagonist. Naloxone and opioid agents, such as morphine, beta-endorphin and [Met(5)]-enkephalin, significantly enhanced and attenuated the stress-induced increase in noradrenaline release in these regions and the stress-induced emotional change, respectively. Two stressful events which predominantly involve emotional factors, i.e., psychological stress and conditioned fear, caused significant increases in noradrenaline release selectively in these three brain regions and these increases were also significantly attenuated by pretreatment with diazepam in a flumazenil reversible manner. Yohimbine, an alpha(2)-adrenoceptor antagonist which caused a marked increase in noradrenaline release in the several brain regions, had an anxiolytic action in the two behavioral tests involving anxiety, i.e., the conditioned defensive burying test and the modified forced swim test. beta-Carbolines, which possess anxiogenic properties, significantly increased noradrenaline release in the hypothalamus, amygdala and locus coeruleus. Taken together, these findings suggest that the increased release of noradrenaline in the hypothalamus, amygdala and locus coeruleus is, in part, involved in the provocation of anxiety and/or fear in animals exposed to stress, and that the attenuation of this increase by benzodiazepine anxiolytics acting via the benzodiazepine receptor/GABAA receptor/chloride ionophore supramolecular complex may be the basic mechanism of action of these anxiolytic drugs. PMID- 11033345 TI - ATP-driven rotation of the gamma subunit in F(1)-ATPase. AB - We present a mechanism for F(1)-ATPase in which hydrolysis of MgATP in the high affinity catalytic site at the alpha/beta interface drives rotation of the gamma subunit via conformational changes in the alpha subunit. During hydrolysis, transition state formation and separation of P(i) from MgADP causes movement of portions of alpha, transmitted via two Arg residues which are hydrogen-bonded to the gamma-phosphate of MgATP, alphaArg376 and betaArg182; the latter is also hydrogen-bonded to interfacial alpha residues between alpha346 and alpha349. Changes in alpha conformation then push on gamma, resulting in rotation. Supporting evidence from the literature and from new data is discussed. PMID- 11033346 TI - Regulation of APP cleavage by alpha-, beta- and gamma-secretases. AB - Proteolytic cleavage of the amyloid protein from the amyloid protein precursor (APP) by APP secretases is a key event in Alzheimer's disease (AD) pathogenesis. alpha-Secretases cleave APP within the amyloid sequences, whereas beta- and gamma secretases cleave on the N- and C-terminal ends respectively. The transmembrane aspartyl protease BACE has been identified as beta-secretase and several proteases (ADAM-10, TACE, PC7) may be alpha-secretases. A number of studies have suggested that presenilins could be gamma-secretases, although this remains to be demonstrated conclusively. Inhibition of beta- and gamma-secretase, or stimulation of alpha-secretase, is a rational strategy for therapeutic intervention in AD. PMID- 11033347 TI - The membrane-bound transferrin homologue melanotransferrin: roles other than iron transport? AB - Melanotransferrin (MTf) is a membrane-bound transferrin (Tf) homologue that is found at high levels in melanoma cells. MTf has many characteristics in common with serum Tf and previous studies have shown that it can bind Fe. This has led to speculation that MTf may be involved in Fe transport. Because Fe is required for a variety of metabolic reactions including ATP and DNA synthesis, MTf could play a role in proliferation. However, recently it has been shown that MTf plays very little role in Fe uptake by melanoma cells, and unlike other Fe transport molecules (e.g. the transferrin receptor), its expression is not controlled by Fe. In the present review the function of MTf is discussed in relation to data suggesting other roles apart from Fe uptake. PMID- 11033348 TI - Single mutation at the intersubunit interface confers extra efficiency to Cu,Zn superoxide dismutase. AB - The Val28-->Gly single mutant at the subunit interface of Cu,Zn superoxide dismutase from Photobacterium leiognathi displays a k(cat)/K(M) value of 1.7x10(10) M(-1) s(-1), twice that of the native enzyme. Analysis of the three dimensional structure indicates that the active site Cu,Zn center is not perturbed, slight structural deviations being only localized in proximity of the mutation site. The enzyme-substrate association rate, calculated by Brownian dynamics simulation, is identical for both enzymes, indicating that the higher catalytic efficiency of the Val28-->Gly mutant is not due to a more favorable electrostatic potential distribution. This result demonstrates the occurrence of an intramolecular communication between the mutation site and the catalytic center, about 18 A away and indicates a new strategy to encode extra efficiency within other members of this enzymatic family. PMID- 11033349 TI - Investigation of the active site of Escherichia coli Cu,Zn superoxide dismutase reveals the absence of the copper-coordinated water molecule. is the water molecule really necessary for the enzymatic mechanism? AB - The active site of the Cu,Zn superoxide dismutase from Escherichia coli in the oxidized Cu(II) state has been studied by nuclear magnetic relaxation dispersion (NMRD), optical and nuclear magnetic resonance spectroscopy. The orientation of some metal ligands is different with respect to all the other Cu,Zn superoxide dismutases. Moreover, NMRD measurements demonstrate the lack of a copper coordinated water molecule. In spite of these differences the enzymatic activity is still high. Azide also binds copper with normal affinity and induces modifications in the active site comparable to those previously observed in the eukaryotic enzymes. Our results suggest that, in this enzyme, the copper coordinated water molecule appears not necessary for the enzymatic reaction. A role for the copper-coordinated water molecule is discussed in the light of recent crystallographic studies. PMID- 11033350 TI - Recombinant C1b domain of PKCdelta triggers meiotic maturation upon microinjection in Xenopus laevis oocytes. AB - The C1 domains are 50 amino acid sequences present in protein kinase C (PKC) isozymes that are responsible for binding of phorbol esters and the lipid second messenger diacylglycerol (DAG). We found that bacterially expressed C1b domain of PKCdelta induces germinal vesicle breakdown (GVBD) when microinjected into Xenopus laevis oocytes. Injection of the C1b domain of PKCdelta significantly enhanced insulin- but not progesterone-induced maturation. Interestingly, the PKCdelta C1b domain markedly synergized with normal Ras protein to induce oocyte maturation when both proteins were co-injected in oocytes. Our results demonstrate that the purified C1b domain of PKCdelta is sufficient to promote meiotic maturation of X. laevis oocytes probably through activation of components of the insulin/Ras signaling pathway. PMID- 11033351 TI - Export of a misprocessed GPI-anchored protein from the endoplasmic reticulum in vitro in an ATP- and cytosol-dependent manner. AB - Strict quality control mechanisms within the mammalian endoplasmic reticulum act to prevent misfolded and unprocessed proteins from entering post-endoplasmic reticulum (ER) compartments. Following translocation into the ER lumen via the Sec61p translocon, nascent polypeptide chains fold and are modified in an environment that contains numerous chaperones and other folding mediators. Recently it has emerged that polypeptides failing to acquire the native state are re-exported from the ER to the cytosol for ultimate degradation by the proteasome ubiquitin system, apparently mediated again via Sec61p. Substrates for this degradation pathway include proteins destined to become glycosyl phosphatidylinositol (GPI)-anchored, but which fail to be processed and retain the C-terminal GPI signal peptide. In order to characterise this process we have used a model GPI-anchored mutant protein, prepro mini human placental alkaline phosphatase (PLAP) W179, which cannot be processed efficiently on account of being a poor substrate for the transamidase which cleaves the GPI signal peptide and adds the GPI anchor in a coupled reaction. In vitro transcription, translation and translocation into canine pancreatic microsomes resulted in ER targeting signal sequence cleavage and formation of prominiPLAP in the ER lumen. We were able to show that prominiPLAPW179 could be exported from the microsomes in a time-dependent manner and that release requires both ATP and cytosol. Export was not supported by GTP, indicating a biochemical distinction from glycopeptide export which we showed recently requires GTP hydrolysis. The process was not affected by redox, unlike several other GPI-anchored model proteins. These data demonstrate that misprocessed proteins can be exported in vitro from mammalian microsomes, facilitating identification of factors involved in this process. PMID- 11033352 TI - Microtubule binding of the drosophila DMAP-85 protein is regulated by phosphorylation in vitro. AB - The phosphorylation of microtubule-associated proteins (MAPs) is thought to be a key factor in the regulation of microtubule (MT) stability. Previously we isolated DMAP-85, a Drosophila MAP shown to be associated with stable MTs. In this work we show that DMAP-85 phosphorylated in cell-free early embryo extracts is released from MTs. MPM-2 antibodies recognize the phosphorylated protein. In vitro, DMAP-85 can be phosphorylated by the mitotic kinase Polo affecting its binding to MTs and creating MPM-2 epitopes on the protein. The results suggest that phosphorylation of DMAP-85 might affect its MT stabilizing activity during early mitotic cycles. PMID- 11033353 TI - In synergy with various cis-acting elements, plant insterstitial telomere motifs regulate gene expression in Arabidopsis root meristems. AB - The telo-box, an interstitial telomere motif, was shown to regulate gene expression in root meristems, in synergy with a cis-acting element involved in the activation of expression of plant eEF1A genes, encoding the translation elongation factor EF1A, and of several ribosomal protein genes. We demonstrate here that the telo-box is also required for transcription activation by two other cis elements present within the promoter of genes encoding the acidic ribosomal protein rp40 and the proliferating cell nuclear antigen respectively. The control of gene expression by telo-boxes during cell cycle progression in Arabidopsis root meristems is discussed. A parallel is drawn with the function of telomeric sequences in Saccharomyces cerevisiae. PMID- 11033354 TI - NADPH:protochlorophyllide oxidoreductase from Synechocystis: overexpression, purification and preliminary characterisation. AB - NADPH:protochlorophyllide oxidoreductase (POR) catalyses the light-dependent reduction of protochlorophyllide to chlorophyllide, a key regulatory reaction in the chlorophyll biosynthetic pathway. POR from the cyanobacterium Synechocystis has been overproduced in Escherichia coli with a hexahistidine tag at the N terminus. This enzyme (His(6)-POR) has been purified to homogeneity and a preliminary characterisation of its kinetic and substrate binding properties is presented. Chemical modification experiments have been used to demonstrate inhibition of POR activity by the thiol-specific reagent N-ethyl maleimide. Substrate protection experiments reveal that the modified Cys residues are involved in either substrate binding or catalysis. PMID- 11033355 TI - Overlap between the ligand recognition properties of the anandamide transporter and the VR1 vanilloid receptor: inhibitors of anandamide uptake with negligible capsaicin-like activity. AB - Some synthetic agonists of the VR1 vanilloid (capsaicin) receptor also inhibit the facilitated transport into cells of the endogenous cannabinoid anandamide (arachidonoylethanolamide, AEA). Here we tested several AEA derivatives containing various derivatized phenyl groups or different alkyl chains as either inhibitors of the AEA membrane transporter (AMT) in intact cells or functional agonists of the VR1 vanilloid receptor in HEK cells transfected with the human VR1. We found that four known AMT inhibitors, AM404, arvanil, olvanil and linvanil, activate VR1 receptors at concentrations 400-10000-fold lower than those necessary to inhibit the AMT. However, we also found three novel AEA derivatives, named VDM11, VDM12 and VDM13, which inhibit the AMT as potently as AM404 but exhibit little or no agonist activity at hVR1. These compounds are weak inhibitors of AEA enzymatic hydrolysis and poor CB(1)/CB(2) receptor ligands. We show for the first time that, despite the overlap between the chemical moieties of AMT inhibitors and VR1 agonists, selective inhibitors of AEA uptake that do not activate VR1 (e.g. VDM11) can be developed. PMID- 11033356 TI - Regulation of tumour necrosis factor alpha mRNA stability by the mitogen activated protein kinase p38 signalling cascade. AB - The translation of tumour necrosis factor alpha (TNFalpha) mRNA is regulated by the stress-activated protein kinase p38, which also controls the stability of several pro-inflammatory mRNAs. The regulation of TNFalpha gene expression in a mouse macrophage cell line RAW264.7 was re-examined using an inhibitor of stress activated protein kinases. Stimulation of these cells with bacterial lipopolysaccharide resulted in stabilisation of TNFalpha mRNA, which was reversed by specific inhibition of p38. An adenosine/uridine-rich element from the TNFalpha 3' untranslated region conferred p38-sensitive decay in a tetracycline regulated mRNA stability assay. Therefore the p38 pathway also controls TNFalpha mRNA turnover. PMID- 11033357 TI - Increased synthesis and decreased stability of mitochondrial translation products in yeast as a result of loss of mitochondrial (NAD(+))-dependent isocitrate dehydrogenase. AB - We have previously demonstrated that the yeast Krebs cycle enzyme NAD(+) dependent isocitrate dehydrogenase (Idh) binds specifically and with high affinity to the 5'-untranslated leader sequences of mitochondrial mRNAs in vitro and have proposed a role for the enzyme in the regulation of mitochondrial translation [Elzinga, S.D.J. et al. (2000) Curr. Genet., in press]. Although our studies initially failed to reveal any consistent correlation between idh disruption and mitochondrial translational activity, it is now apparent that compensatory extragenic suppressor mutations readily accumulate in idh disruption strains thereby masking mutant behaviour. Now, pulse-chase protein labelling of isolated mitochondria from an Idh disruption mutant lacking suppressor mutations reveals a strong (2-3-fold) increase in the synthesis of mitochondrial translation products. Strikingly, the newly synthesised proteins are more short lived than in mitochondria from wild-type cells, their degradation occurring with a 2-3-fold reduced half-life. Enhanced degradation of translation products is also a feature of yeast mutants in which tethering/docking of mitochondrial mRNAs is disturbed. We therefore suggest that binding of Idh to mitochondrial mRNAs may suppress inappropriate translation of mitochondrial mRNAs. PMID- 11033358 TI - Abscisic acid plasmalemma perception triggers a calcium influx essential for RAB18 gene expression in Arabidopsis thaliana suspension cells. AB - Pretreatment of Arabidopsis thaliana suspension cells with impermeant calcium chelator EGTA inhibited the ABA-induced RAB18 gene expression. However, extracellular calcium alone, up to 10 mM, did not trigger RAB18 expression. Spectrofluorimetric extracellular Ca(2+) measurement with Fluo-3 showed a fast, within 1 min, Ca(2+) influx associated with outer plasmalemma ABA perception. In the presence of the Ca(2+) blockers Cd(2+) and Ni(2+), RAB18 expression was suppressed. Pimozide and fluspirilene inhibited Ca(2+) influx and ABA-induced RAB18 expression. Thus we demonstrated the involvement of specific Ca(2+) influx in the ABA signaling sequence leading to RAB18 expression. PMID- 11033359 TI - The DNA replication-related element (DRE)-DRE-binding factor (DREF) system may be involved in the expression of the Drosophila melanogaster TBP gene. AB - The TATA box binding protein (TBP) is a general transcription factor required for initiation by all three eukaryotic RNA polymerases. Previously, we found that the promoter region of the Drosophila melanogaster TBP gene contains three sequences similar to the DNA replication-related element (DRE) (5'-TATCGATA). In the present study, we found that the DRE-like sequences are also present in the promoter of the Drosophila virilis TBP gene, suggesting a role for these sequences in TBP expression. Band mobility shift assays revealed that oligonucleotides containing sequences similar to the DRE of D. melanogaster TBP gene promoter form specific complexes with a factor in a Kc cell nuclear extract and with recombinant DRE-binding factor (DREF). Furthermore, these complexes were either supershifted or diminished by monoclonal antibodies to DREF. Transient luciferase assays demonstrated that induction of mutations in two DRE-related sequences at positions -223 and -63 resulted in an extensive reduction of promoter activity. Thus, the DRE-DREF system appears to be involved in the expression of the D. melanogaster TBP gene. PMID- 11033360 TI - Differential modulation of nitric oxide production by curcumin in host macrophages and NK cells. AB - Curcumin, the yellow pigment from Curcuma longa, has been shown to possess tumoricidal activity. We have earlier reported the induction of apoptosis in AK 5, rat histiocytic cells by curcumin leading to the inhibition of tumor growth in vivo. In this study we have observed differential activation status in host macrophages and NK cells induced by curcumin during the spontaneous regression of subcutaneously transplanted AK-5 tumors. Closer scrutiny of the cytokine profile and nitric oxide (NO) production by immune cells showed an initial downregulation of Th1 cytokine response and NO production by macrophages, and their upregulation in NK cells, which picked-up upon prolonged treatment with curcumin, culminating in a stronger tumoricidal effect. These studies suggest that the host macrophages and NK cells play an important modulatory role in the remission of AK-5 tumor. PMID- 11033361 TI - Variation in barnacle recruitment over small scales: larval predation by adults and maintenance of community pattern. AB - Over small spatial scales, variation in the density of settlers of benthic sessile species is the result of interactions among larval behavior, local hydrodynamic conditions, and the physical, chemical and biological characteristics of the benthic habitat. It has been shown repeatedly that adult benthic filter-feeders can consume larvae of their own and other species, but their effects on the distribution and abundance of recruits have rarely been demonstrated under natural conditions in the field, particularly on hard substrata. Here we experimentally quantified the effect of the large intertidal barnacle, Semibalanus cariosus (Pallas), on the density of recruits of three common barnacle species. The experiments were conducted at the peak of the barnacle recruitment season over three successive years, on the west coast of San Juan Island, Washington. A persistent and well documented community pattern in the mid intertidal zone of the study site is a sparse bed of adult S. cariosus with bare rock spaces essentially devoid of small barnacles among the large individuals. Field experiments consisted of small areas from which either all adult S. cariosus were killed leaving the shells attached to the rock, or live adult barnacles were left intact. Our results showed that over small spatial scales of a few to tens of centimeters, the large barnacle S. cariosus can interfere and significantly reduce net settlement and recruitment of conspecific as well as other barnacle species. Between 65 and 100% reduction in settlement could be attributed to larval predation by adults, as implied by barnacle settlement patterns on different treatments and by the presence of nauplius larvae in cirri and stomach contents of S. cariosus. The negative effect on barnacle settlement was consistent between years of relatively low barnacle recruitment, which appears to be the most common situation at the study site, but it disappeared on a year of unusually high recruitment, when settling larvae seem to have swamped the filtration ability of adult S. cariosus. The different barnacle species displayed contrasting settlement patterns on bare rock and on the lateral shells of the large barnacles, which appear to be a result of differences in larval behavior. Comparisons against the relative availability of these substrata in the experimental plots suggested that larvae of different species sample the benthic microhabitat in very different ways. PMID- 11033362 TI - The role of mounds in promoting water-exchange in the egg-tending burrows of monogamous goby, Valenciennea longipinnis (Lay et Bennett). AB - Valenciennea longipinnis spawns monogamously in a burrow. After spawning, the paired female constructs a conspicuous mound on the burrow by carrying and piling up substratum-derived materials while the male tends eggs in the burrow until hatching occurs. In this study, the mounds of V. longipinnis were tested in the field to confirm their function of promoting water-exchange in the burrow, and their ecological role was examined in relation to egg care by the male. The mound of V.longipinnis promoted water-exchange in the burrow, contributing to the provision of external oxygenated sea water into the burrow. Therefore, dissolved oxygen (DO) concentrations in the burrow with a mound were significantly higher than those without a mound. Although male egg-tending behaviour (e.g., fanning) may also promote water-exchange in the burrow, the water-exchange appeared to depend mainly on the hydrodynamic effect. Removals of the mound and paired female on the day of spawning led to high rates of egg-desertion by males. Since the frequency and time of fanning increase with a decrease of DO concentration in the burrow, the egg-desertion may result from an increased parental cost to males due to the decrease of water-exchange without a mound. This was supported by the fact that the DO concentrations on the day after mound removal were significantly lower in the egg-deserted burrows (measured before desertions) than in burrows not deserted by the male. Moreover, removals of paired females only also led to higher desertion rates. After removal of the female, the mound gradually collapsed by wave action and other factors, and the surface of the mound was covered with planktonic materials. Such a mound of poor quality may provide little water-exchange, which may lead to the egg-desertion by males. These results indicated that mound maintenance by females during the egg-tending period has an important role in the success of parental care by males. PMID- 11033363 TI - Mortality, behavior and the effects of predators on the intertidal distribution of littorinid gastropods. AB - Predators can affect the vertical distribution of mobile intertidal invertebrates in two ways: they can (1) cause greater mortality of prey at certain intertidal levels, and (2) induce prey to seek safer intertidal areas. In this study, we investigate whether low-intertidal and subtidal predators affect the intertidal distribution of two congeneric species of small herbivorous gastropods of northeastern Pacific shores, Littorina sitkana Philippi 1846, and L. scutulata Gould 1849. In particular, we tested the hypothesis that predators affect the distribution of these snails by inducing them to seek higher and safer intertidal areas. On a wave-sheltered shore in Barkley Sound, British Columbia, L. sitkana and L. scutulata were both killed by predatory crabs (e.g., Cancer productus) more frequently when tethered near the lower limit of their intertidal distribution ( approximately 1 m) than when tethered where they were most common ( approximately 2.5 m), suggesting that high mortality rates are partly responsible for the lower-limit of these snails' intertidal distribution. However, two field mark-recapture experiments indicated that the snails' behavioral response to predation risk also influences their distribution. In the first experiment, snails from the 2.5-m level (low risk) transplanted to the 1.0 m level (high risk) displayed a strong and consistent tendency to move shoreward, especially L. sitkana, some traveling 10-15 m in 2-3 days to regain their original level. These shoreward movements were especially precise in the northern part of the study area, where predation rates on tethered snails were greatest. Furthermore, larger more vulnerable snails were more strongly oriented shoreward than smaller individuals, indicating that antipredator behavior might also contribute to intertidal size gradients in these species. In the second mark recapture experiment, we manipulated predation risk using small cages and found that snails exposed to the odors of C. productus crabs foraging on conspecific and heterospecific snails displayed more precise (L. sitkana and L. scutulata) and longer (L. sitkana) shoreward movements than snails held in control conditions. These results provide the first experimental evidence that antipredator behavior may contribute to the intertidal distribution patterns of littorinids. PMID- 11033364 TI - Effects of temperature and salinity on nitrogenous excretion by Litopenaeus vannamei juveniles. AB - Excretion rates of ammonia-N, nitrite-N, nitrate-N, and dissolved organic nitrogen (DON) for juvenile Litopenaeus vannamei (3.85+/-0.83 g) were quantified in response to nine different combinations of temperature (24, 28, and 32 degrees C) and salinity (10, 25, and 40 ppt) under laboratory conditions. Results indicated that L. vannamei is ammonotelic, with ammonia-N accounting for 61.9 84.3% of total nitrogen (TN) excretion. There were significant effects of temperature and salinity, but no significant interaction between them, on ammonia N excretion rate (R(AN)). R(AN) increased with increasing temperature, over the interval 24-32 degrees C. R(AN) was lower at 25 ppt than at 10 and 40 ppt, at all temperatures. DON excretion rate (R(DON)) was not significantly influenced by either temperature or salinity; the overall mean R(DON) was about 5.24 ug-N g -1 h -1. However, the percentages of DON in TN (P(DON)) varied from 15.4 to 36.4% under the various temperature-salinity combinations. P(DON) at 28 and 32 degrees C was significantly lower than at 24 degrees C, and P(DON) at 10 ppt was significantly lower than at 25 and 40 ppt. Only very small amounts of nitrogen were excreted by L. vannamei as nitrite-N and nitrate-N. PMID- 11033365 TI - Postingestive selection in the sea scallop, Placopecten magellanicus (Gmelin): the role of particle size and density. AB - Suspension-feeding bivalves can influence the energy value of their food supply by particle selection at various stages from particle clearance to production of feces. Previous workers have found that some bivalve species (Mercenaria mercenaria, Mytilus edulis) are capable of postingestive selection within the stomach. Few studies, however, have attempted to isolate the factors that influence postingestive selection. In this study, we examined the ability of the sea scallop Placopecten magellanicus to select particles within the stomach on the basis of physical properties. Scallops were presented with a mixture of three sizes of beads (5, 10 and 20 um) or with a mixture of beads of different densities (1.05 g ml(-1) and 2.5 g ml(-1)). We demonstrate that P. magellanicus can distinguish between particles of different sizes and densities, retaining larger particles (20 um) longer than smaller ones (5 um) and lighter particles longer than denser ones. This ability to reject small, dense particles may benefit the scallop by reducing the amount of energy expended attempting to digest poor quality particles such as silt. This paper presents the first quantitative analysis of the effect of particle size and density on particle processing within intact bivalves. PMID- 11033366 TI - Growth and tissue composition as a function of feeding history in juvenile cephalopods. AB - We present the results of a series of experiments that examined the effect of feeding history on the growth and tissue composition of juveniles of two tropical cephalopods; the squid Sepioteuthis lessoniana and the cuttlefish Sepia elliptica. Juveniles were reared in individual containers for between 35 and 42 days at different ration levels, three ration levels for the squid and two levels for the cuttlefish. Although differences in ration were sufficient to cause different growth rates, both in body length and mass, the effects on tissue composition were less definitive. Sepioteuthis juveniles on the highest rations had higher concentrations of water, but no difference in lipid, carbohydrate or protein when compared with their lower ration siblings. In the case of juvenile cuttlefish no difference in tissue composition was detected between the two ration levels. RNA:protein ratios were also determined for the juveniles to provide an estimate of instantaneous growth. A significant correlation was found between body size and RNA:protein ratio in the squid; those juveniles that ate more had higher RNA:protein ratios than lower ration individuals. Significantly, the juvenile cuttlefish showed no relationship between growth rate and RNA:protein ratios, which means that we are unable to use this measure to estimate the growth rates of wild individuals. In conclusion, ration level did affect growth rates and food availability is an important factor in modifying growth rates of wild individuals. However, we could not find, at the individual level, an index or measure that could be used to explain the variability of observed differences in growth rates as a function of nutritional history. PMID- 11033367 TI - Benthic predator-prey interactions: evidence that adult Monoporeia affinis (Amphipoda) eat postlarval Macoma balthica (Bivalvia). AB - Predation by adults of the amphipod Monoporeia affinis on the plantigrade postlarval stage of the bivalve Macoma balthica was studied in the laboratory. We confirmed that M. affinis consumes small M. balthica. Amphipods offered 14C labelled postlarvae took up the radioactive tracer, while those presented Rhodamine B-stained postlarvae acquired gut contents fluorescing strongly in orange, whereas control amphipods did not. Both labelling methods proved convenient to use in laboratory experiments, and are particularly useful when organisms lack structures that can be easily identified after being ingested, or when cross-over reactions may bias the results of immunoassays. The results reported here support the conclusion from earlier studies that predation by M. affinis on M. balthica can affect population dynamics of M. balthica and is likely to be an important structuring factor in the low-diversity benthic macrofauna community of the Baltic Sea. PMID- 11033368 TI - Effect of periodical starvation on the life history of Brachionus plicatilis O.F. Muller (Rotifera): a possible strategy for population stability. AB - To estimate the changes in the life history of the rotifer Brachionus plicatilis O.F. Muller under starvation, we carried out an individual culture and determined the effects of periodical food deprivation on its asexual reproductive characteristics such as lifespan, reproductive period, age at first egg and offspring production, and lifetime fecundity (total number of offspring produced in her lifetime). Rotifers were fed for 1-3 h daily, and were then starved until the next day. Control animals were fed throughout their lifespan. Starved rotifers matured and produced their first offspring at an older age than the control animals. The periodical starvation resulted in a decrease in the lifetime fecundity to less than half that of the non-starved control. The reproductive period and lifespan were 2-3 times longer in the starved animals than in the control animals. The negative relationship between lifespan and lifetime fecundity is interpreted as a trade-off in an alternative life-history strategy of rotifers under starved conditions. The great decrease in fecundity and extension of lifespan enables rotifers to compensate to keep the population in equilibrium. PMID- 11033369 TI - Why put new wine in old bottles: the need for a biopsychosocial approach to the assessment, treatment, and understanding of unexplained and explained symptoms in medicine. PMID- 11033370 TI - The role of personality in the development and perpetuation of chronic fatigue syndrome. AB - OBJECTIVES: Qualitative evidence suggests that personality may have special relevance to the predisposition, precipitation and perpetuation of chronic fatigue syndrome (CFS). This study compares three dimensions of personality - perfectionism, self-esteem, and emotional control in the personality profiles of CFS patients (N=44) and a control group (N=44) without a history of CFS, matched for age and gender. METHODS: Participants were assessed on the MPS [Frost RO, Marten P, Lahart C, Rosenblate R. The dimensions of perfectionism. Cognit Ther Res 1990;14:449-468.]; the Rosenberg Self-Esteem Scale [Rosenberg M. Society and the Adolescent Self-image. Princeton, NJ: Princeton Univ Press, 1965.]; the Courtauld Emotional Scale [Watson M, Greer S. Development of a questionnaire measure of emotional control. J Psychosom Res 1983;27:299-305.] and the Marlowe Crowne Social Desirability Scale [Crowne DP, Marlowe D. A new scale of social desirability independent of psychopathology. J Consult Psychol 1960;24:349-354.]. RESULTS: Analyses revealed that the CFS group reported higher levels than the control group on the Total Perfectionism score and Doubts about Actions and the Concern over Mistakes subscales. Furthermore, the CFS group also reported lower self-esteem than the control group. No difference between the two groups was found on the dimensions of emotional control and social desirability response bias. CONCLUSION: A developmental model of CFS, which considers the predisposing, precipitating, and perpetuating factors that may account for the course of the disorder irrespective of etiology, is proposed. In the context of the results, recommendations for practice and future research are discussed. PMID- 11033372 TI - Mood state as a predictor of neuropsychological deficits following cardiac surgery. AB - OBJECTIVES: mood disorders and neuropsychological deficits are both commonly reported occurrences after cardiac surgery. We examined the relationship between mood state and postoperative cognitive deficits in this population. METHODS: assessments of neuropsychological functions and mood state (depression, anxiety, stress scales; DASS) were performed preoperatively and postoperatively on 147 patients undergoing cardiac surgery. RESULTS: the incidence of preoperative depression, anxiety, and stress symptomatology was 16%, 27%, and 16%, respectively. The incidence of postoperative anxiety symptomatology significantly increased to 45% (p<0.001), while the incidence of depression and stress symptomatology remained stable (19% and 15%, respectively; ns). Changes in mood state did not influence changes in neuropsychological performance. Preoperative mood was a strong predictor of postoperative mood, and was related to postoperative deficits on measures of attention and memory. CONCLUSIONS: an assessment of preoperative mood is critical in identifying patients at risk of postoperative mood disorders and neuropsychological deficits. Measures assessing somatic manifestations of anxiety may not be suitable for a surgical population. PMID- 11033371 TI - Stressors, personality traits, and coping of Gulf War veterans with chronic fatigue. AB - OBJECTIVES: preliminary surveys of Persian Gulf veterans revealed a significant prevalence of self-reported symptoms consistent with chronic fatigue syndrome (CFS). The purpose of this study was to compare self-reported life stressors, combat, and chemical exposures, personality and coping between Gulf War veterans with CFS and healthy veterans. METHODS: following a complete physical, psychiatric, and neuropsychological evaluation, 45 healthy veterans, 35 veterans with CFS and co-morbid psychiatric disorder, and 23 veterans with CFS and no co morbid psychiatric disorder completed questionnaires assessing war and non-war related life stressors, self-reports of environmental exposure (e.g. oil well fires, pesticides), personality, and coping. RESULTS: measures of personality, self-reported combat and chemical exposures, and negative coping strategies significantly differentiated healthy veterans from those with CFS. CONCLUSION: a biopsychosocial model of veterans' illness was supported by the fact that personality, negative coping strategies, life stress after the war, and environmental exposures during the war were significant predictors of veterans' current physical function. PMID- 11033373 TI - Coping strategies in twins with chronic fatigue and chronic fatigue syndrome. AB - OBJECTIVES: Individuals with chronic fatigue and chronic fatigue syndrome (CFS) face debilitating symptoms as well as stressful life situations that may result from their condition. The goal of this study was to examine the coping strategies used by fatigue-discordant twin pairs. METHODS: We utilized a co-twin design to assess how twin pairs discordant for chronic fatigue and CFS cope with stress. All twin pairs were administered the Revised Ways of Coping Checklist. RESULTS: Overall, the pattern of coping strategies was similar for fatigued and non fatigued twins. However, twins with chronic fatigue or CFS utilized more avoidance strategies than their non-fatigued counterparts; those with chronic fatigue also used more avoidance relative to problem-focused coping than their co twins. CONCLUSIONS: These results suggest that while fatigue-discordant twins generally exhibit similar behavior patterns in order to cope with stress, there may be an association between fatigue and avoidance coping. Future research should focus on the role of avoidance and its relationship to fatiguing illnesses. PMID- 11033374 TI - Athens Insomnia Scale: validation of an instrument based on ICD-10 criteria. AB - OBJECTIVES: To describe and validate the Athens Insomnia Scale (AIS). METHODS: The AIS is a self-assessment psychometric instrument designed for quantifying sleep difficulty based on the ICD-10 criteria. It consists of eight items: the first five pertain to sleep induction, awakenings during the night, final awakening, total sleep duration, and sleep quality; while the last three refer to well-being, functioning capacity, and sleepiness during the day. Either the entire eight-item scale (AIS-8) or the brief five-item version (AIS-5), which contains only the first five items, can be utilized. The validation of the AIS was based on its administration to 299 subjects: 105 primary insomniacs, 144 psychiatric patients and 50 non-patient controls. RESULTS: Regarding internal consistency, for both versions of the scale, the Cronbach's alpha was around 0. 90 and the mean item-total correlation coefficient was about 0.70. Moreover, in the factor analysis, the scale emerged as a sole component. The test-retest reliability correlation coefficient was found almost 0.90 at a 1-week interval. As far as external validity is concerned, the correlations of the AIS-8 and AIS-5 with the Sleep Problems Scale were 0.90 and 0.85, respectively. CONCLUSION: The high measures of consistency, reliability, and validity of the AIS make it an invaluable tool in sleep research and clinical practice. PMID- 11033375 TI - The relationship of alexithymia characteristics to dreaming. AB - OBJECTIVE: In two studies, we tested two hypotheses about the relationship of alexithymia to dreaming; that dreams of alexithymic people are barren and rarely recalled, and that the dreams are unregulated and nightmarish. METHODS: Study 1 was a retrospective survey of dreaming among several hundred young adults, and Study 2 was a 1-week, prospective diary study of 153 young adults in which recall, content, and length of dreams were assessed. RESULTS: Across both studies, the externally oriented thinking (EOT) facet of the Toronto Alexithymia Scale (TAS)-20 correlated with different dream characteristics than the difficulty identifying feelings (DIF) and difficulty describing feelings (DDF) facets, even after statistically controlling for the other facets. Greater EOT was related to an increased frequency of nights without dream recall, having shorter dreams, having dreams rated as boring and lacking vividness, and not believing in the importance of dreams. In contrast, greater DIF or DDF was related to an increased frequency of nights with disturbing dreams, and having dreams rated as bizarre and aggressive. CONCLUSION: We find support for both hypotheses, but different facets of the multidimensional alexithymia construct account for the two types of dream reports. PMID- 11033376 TI - Mediators of stress effects in inflammatory bowel disease: not the usual suspects. AB - Empirical efforts to prove or disprove an association between stress and the course of inflammatory bowel disease (IBD) have had inconsistent results. Direct study of mediators of stress-related physiological processes may clarify this important area. METHODS: candidate mediators were selected based on evidence that they have a role in the pathophysiology of IBD. Medline searches for original articles addressing each putative mediator and psychological stress were conducted. Articles were reviewed with goals of synthesis and hypothesis generation. RESULTS: there is evidence that substance P (SP), vasoactive intestinal protein (VIP), tumour necrosis factor alpha (TNFalpha), oxidant molecules, endogenous glucocorticoids and heat shock proteins (HSPs) are involved in the stress response. DISCUSSION: two principles emerge which should inform efforts to investigate stress in IBD. First, stress effects are regulated by highly interdependent systems. Second, the effects of mediators are highly specific to the location of their activity, and so, investigations in IBD are likely to require direct investigation of inflamed and unaffected gut tissue. PMID- 11033377 TI - Construct validation of the hospital anxiety and depression scale with clinical populations. AB - OBJECTIVE: the hospital anxiety and depression scale (HADS) attempts to measure anxiety and depression without confounding by somatic symptoms of physical disorder, and is widely used for this purpose. This paper addresses three questions about the validity of the HADS concerning its independence of physical symptoms, the extent to which its items robustly measure the identified constructs with varying clinical populations and situations, and its capacity to differentiate anxiety and depression. METHODS: data from patients with breast disease, myocardial infarction (MI), and stroke were examined using factor analytic and psychometric analyses. RESULTS: using exploratory factor analysis in patients with breast disease, 13 of the 14 HADS items fell on a psychological factor and loadings on the psychological factor were higher than loadings on the somatic factor for all items. The HADS showed high levels of internal consistency and there was little evidence that removing items would improve it. Confirmatory factor analyses (CFA) in MI and stroke groups confirmed the separation of anxiety and depression. Analyses indicated items, which were performing poorly for these clinical groups. CONCLUSIONS: there was support for the validity of the HADS for all three questions. However, there were some evidences of individual items performing poorly. Given the ease of administration and the acceptability of this measure to ill or weak respondents, the HADS continues to perform satisfactorily. PMID- 11033378 TI - Panic disorder and idiopathic cardiomyopathy. AB - OBJECTIVE: This study investigated whether, among a population of cardiac patients, there is a preferential association between idiopathic cardiomyopathy (CMP) and Panic Disorder (PD). METHODS: A total of 93 patients with cardiac failure, 50 of them with CMP, 43 with other cardiac diseases, underwent a standard psychiatric examination using the MINI neuropsychiatric interview. RESULTS: While half of the subjects met the criteria for a psychiatric disorder, PD was no more prevalent in the CMP group compared to other patients. CONCLUSION: This study confirms the high prevalence of psychiatric pathology, particularly anxiety, mood, and alcohol-related disorders, in patients with cardiac failure. However, previous findings on the specific association between CMP and PD could not be replicated. PMID- 11033379 TI - Psychiatric disorders in patients with blepharospasm - a reactive pattern? AB - OBJECTIVE: The purpose of this study was to assess the life-time prevalence of all major psychiatric disorders in patients suffering from blepharospasm. METHOD: A total of 31 consecutive patients with blepharospasm attending the Department of Neurology were interviewed at the Department of Psychiatry at the University of Vienna. Patients had been submitted to standard neurological diagnostic procedures, psychiatric diagnoses were made with the help of the SCID, functional impairment was assessed by the General Assessment of Functioning Scale (GAF). RESULTS: A current or life-time psychiatric diagnosis was made for 22 patients (71%). The most frequent disorders were depressive disorders, mainly major depression (five patients, 16.1%), secondary dysthymia (six patients, 19.3%), and recurrent major depression (five patients, 16.1%). A diagnosis of simple phobia was made for seven patients (22.5%), for obsessive-compulsive disorder in three patients (9.6%). The mean GAF score of our sample was 63.1%. CONCLUSION: In contrast to previously published results, we did not find a high rate of a single specific disorder or patterns in our study sample, though by the inclusion of life-time diagnostic criteria, the majority of patients fulfilled criteria for at least one diagnosis. This might indicate the considerable negative impact of blepharospasm on the patients' lives. PMID- 11033380 TI - Is sleep-disordered breathing an independent risk factor for hypertension in the general population (13,057 subjects)? AB - OBJECTIVES Sleep-disordered breathing has been hypothesized to have a close relationship with hypertension but previous studies have reported mixed results. This is an important health issue that requires further clarification because of the potential impact on the prevention and control of hypertension. METHODS: The relationship between hypertension and three forms of sleep-disordered breathing (chronic snoring, breathing pauses and obstructive sleep apnea syndrome (OSAS)) was assessed using representative samples of the non-institutionalized population of the UK, Germany and Italy (159 million inhabitants). The samples were comprised of 13,057 individuals aged 15-100 years who were interviewed about their sleeping habits and their sleep symptoms over the telephone using the Sleep EVAL system. RESULTS: OSAS was found in 1.9% (95% CI: 1.2% to 2.3%) of the UK sample, 1.8% (95% CI: 1.4% to 2.2%) of the German sample and 1.1% (95% CI: 0.8% to 1.4%) of the Italian sample. OSAS was an independent risk factor (odds ratio (OR): 9.7) for hypertension after controlling for possible confounding effects of age, gender, obesity, smoking, alcohol consumption, life stress, and, heart and renal disease. CONCLUSIONS: Results from three of the most populated countries in Western Europe indicate that OSAS is an independent risk factor for hypertension. Snoring and breathing pauses during sleep appeared to be non-significant predictive factors. PMID- 11033381 TI - beta(2)-Adrenoceptors and ventricular fibrillation. AB - beta-Adrenoceptor antagonists significantly reduce the incidence of sudden cardiac death in patients with contractile dysfunction. Contractile dysfunction is associated with a decline in beta(1)-adrenoceptors, no change in the number of beta(2)-adrenoceptors, and an increased responsiveness to beta(2)-adrenoceptor stimulation. Selective beta(2)-adrenoceptor blockade prevents ventricular fibrillation in a canine model of sudden cardiac death. Cardiac beta(2) adrenoceptor stimulation increases L-type Ca(2+) currents, but unlike beta(1) adrenoceptor stimulation, it fails to elicit phospholamban phosphorylation. Restoration of resting diastolic [Ca(2+)] following beta(2)-adrenoceptor-mediated increases in Ca(2+) influx is more dependent on Na(+)/Ca(2+) exchange, which generates an arrhythmogenic transient inward current that can trigger ventricular fibrillation. PMID- 11033382 TI - Mechanisms involved in the hemodynamic alterations in congestive heart failure as a basis for a rational pharmacological treatment. AB - Congestive heart failure is a complex syndrome and one of the major cardiological problems of our time. It is characterized by an important neurohumoral activation to compensate for the reduction of cardiac output and blood pressure, that worsens the prognosis with time. The aim of the treatment is focused on how to improve the quality of life and how to prolong survival. Usually, treatment, either symptomatic or directed to control the neuroendocrine compensatory changes, is necessary. The drugs currently used are angiotensin-converting enzyme inhibitors, diuretics, digoxin, and beta-adrenoceptor agonists. In addition, new drugs, such as angiotensin II receptor antagonists, beta-adrenoceptor antagonists, ibopamine, Ca(2+) antagonists, neutral endopeptidase inhibitors, vasopressin antagonists, Ca(2+)-sensitizers with cyclic AMP-dependent or independent mechanisms, and endothelin antagonists, are also being used. PMID- 11033383 TI - The dose-response relationship in phase I clinical trials and beyond: use, meaning, and assessment. AB - Knowledge of the relationships among dose, drug concentration in blood, and clinical response is important for the safe and effective use of drugs in individual patients. Recently, pharmacokinetic-pharmacodynamic modeling has been taking an increasingly important place in clinical pharmacology because of its role in the determination of the optimal dosage of a new drug. Its primary objective is also to identify the characterization and prediction of the time course of drug effects under physiological and pathological conditions. Dose response studies are useful in Phase I for assessing drug tolerance and safety, and invaluable in Phase II for characterizing drug efficacy. Apart from the confirmation of efficacy, the acquired information may help to investigate the shape and location of the dose-response curve, the choice of an appropriate therapeutic starting dose, the identification of optimal strategies for individual dose adjustments, and the determination of a maximal dose beyond which additional benefit is unlikely to be obtained. Recent development of pharmacodynamic models such as the mechanism-based indirect effect model may permit the identification of the physiological component of drug action that is affected by disease, other medications, gender, and other variables. Assessment of dose response should be an integral component of drug development, with studies designed to assess dose response an inherent part of establishing the safety and efficacy of the drug. Drug development can be enhanced with a good understanding of dose-response characteristics and ultimately the benefit/risk ratio of a drug. PMID- 11033384 TI - Behavioural effects of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor antagonists and their relevance to substance abuse. AB - This review presents some of the work that has been carried out to investigate the behavioural effects of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-receptor antagonists in animal models of substance abuse. Many of the studies have been conducted in light of current ideas that emphasise the analogous role of glutamatergic mechanisms in synaptic plasticity and long-term behavioural adaptation to drugs. Experiments on behavioural sensitisation indicate that whereas N-methyl-D-aspartate receptors are involved in induction, AMPA-receptors may mediate expression of the established response. In this regard, an important factor may be the degree of drug-environment conditioning. Thus, studies of the effects of AMPA-receptor antagonists on conditioned behaviours are reviewed here. Relatively few studies on the effects of AMPA receptor antagonists on primary reinforcement from self-administered drugs and the subjective effects of drugs have been carried out, but a profile that contrasts with that of the N-methyl-D-aspartate antagonists appears to be emerging. Studies of withdrawal from opioids suggest that whilst AMPA-receptor antagonists may not be able to prevent tolerance or dependence from developing, they may ameliorate both the physical and emotional consequences of withdrawal. Overall, the AMPA-receptor antagonists may represent a promising new approach for treating the consequences of drug abuse. However, as results are often complicated by the use of the less-selective compounds, it will be important to use better tools in future studies. PMID- 11033386 TI - Erratum PMID- 11033385 TI - Kallikrein and kinin receptor genes. AB - Recent evidence increasingly supports the view that kinins exercise an important regulatory control in inflammation and in the growth and proliferation of cancer cells. The induction of tissue kallikrein (TK) gene results in either increased or new expression of this protease, resulting in an increased capacity to form kinins. The cellular actions of kinins are initiated and controlled by kinin B1 and B2 receptors. This review collates in detail current knowledge on the molecular profile and status of TK (hKLK1, hKLK2, and hKLK3) and the kinin B1 and B2 receptor genes. The development of TK inhibitors, as well as kinin receptor antagonists, for use in immune-modulated disorders and in tumours may provide a new generation of drugs of therapeutic value. PMID- 11033387 TI - Conquering capsule opacification. PMID- 11033388 TI - Scleral fixation technique using 2 corneal tunnels for a dislocated intraocular lens. AB - Several techniques are used to reposition dislocated intraocular lenses (IOLs). Most place a suture loop around the end of the haptic. However, in cases of a dislocated capsular bag containing the IOL, a dislocated IOL with a large haptic, or a miotic pupil, it is not easy to see the haptic ends to place the suture loop. We describe a scleral fixation technique that uses 2 corneal tunnels. A double-armed 10-0 polypropylene suture loop can be introduced through 1 corneal tunnel and placed around any accessible part of the haptic with the help of a bent 26 gauge needle. This modified technique is an easy and effective way to reposition the IOL. PMID- 11033389 TI - Early clinical experience using custom excimer laser ablations to treat irregular astigmatism. AB - PURPOSE: To assess the viability of custom excimer laser ablations for treating irregular astigmatism. SETTING: Single-center prospective study of a new custom ablation technique. METHODS: Twelve patients received 15 custom ablations for irregular astigmatism resulting from keratoconus, penetrating keratoplasty for keratoconus, prior decentered laser in situ keratomileusis, or incisional refractive surgery. Follow-up ranged from 6 weeks to 14 months. Initially, the laser beam was manually decentered; later, the Contoured Ablation Patterns (CAP) method (VISX, Inc.) was used to automatically decenter the ablation over the corneal elevation. RESULTS: Results are presented in a case-by-case fashion. In the manual decentration group, the uncorrected visual acuity (UCVA) was 20/50 or better in 9 of 11 eyes (81.8%) and 20/40 or better in 7 eyes (63.6%). Surgery resolved or decreased visual symptoms when present. The best corrected visual acuity (BCVA) was maintained or improved in all eyes. Persistent .05). CONCLUSIONS: Psychophysical assessments may be a more sensitive indicator of decreases in visual performance following excimer laser refractive surgery. The attempted level of correction of spherical equivalent and astigmatism may adversely affect early scotopic visual performance. Decreases in visual performance are temporary, return to normal by 12 months, and remain stable 24 months following PARK. PMID- 11033394 TI - Correlation between intraocular pressure and corneal stromal thickness after laser in situ keratomileusis. AB - PURPOSE: To assess the relationship between intraocular pressure (IOP) and corneal stromal thickness following laser in situ keratomileusis (LASIK) for myopia or myopic astigmatism. SETTING: Private clinic in Turkey. METHODS: The database of patients who had LASIK was retrospectively reviewed. Preoperative and postoperative IOP and corneal thickness were measured in both eyes of 353 patients by noncontact tonometry and ultrasonic pachymetry, respectively. The correlation between changes in IOP and corneal thickness was evaluated by regression analysis. RESULTS: Laser in situ keratomileusis was associated with a mean decrease in pachymetry of 46.7 microm +/- 28.9 (SD) and a mean decrease in IOP of 2.8 +/- 2.1 mm Hg. There was a significant correlation between the decreases in IOP and pachymetry (P <.001). CONCLUSION: The decrease in IOP after LASIK was related to the decrease in corneal stromal thickness, although there may be other contributing factors. PMID- 11033393 TI - Clinical characteristics of lamellar channel deposits after implantation of intacs. AB - PURPOSE: To report the frequency, location, and severity of biomicroscopically evident intrastromal deposits that may accumulate in the lamellar channel after implantation of Intacs corneal ring segments. SETTING: United States Food and Drug Administration multicenter clinical trial conducted at 10 sites. METHODS: Two-year follow-up data from a phase III clinical trial (N = 359) were reviewed. Examiners at 10 study sites described the location and severity (density) of deposits, grading severity on a standardized scale from absent (0) to severe (4). At 24 months, the incidence of deposits was 74% overall and 61%, 73%, and 89% with the 0.25, 0.30, and 0.35 mm Intacs, respectively (P <.001). The incidence increased sharply at first and more slowly after 6 months. RESULTS: At 12 months, deposits were located along the inner curvature of the segments in 47% of patients, along the outer curvature in 5%, along both inner and outer curvatures in 38%, and anterior to the segments in 10%. The severity of the deposits increased with segment thickness (P <.001 at 6, 12, and 24 months). CONCLUSIONS: Lamellar channel deposits occurred frequently after Intacs implantation. The incidence and density of deposits increased with segment thickness and duration of implantation. The presence of this material did not result in alteration of the optical performance of Intacs or anatomical or physiological corneal deterioration. PMID- 11033395 TI - Long-term results of multizone photorefractive keratectomy for myopia of -6.0 to 10.0 diopters. AB - PURPOSE: To evaluate the 4 year refractive outcome of multizone photorefractive keratectomy (PRK) in eyes with high myopia. SETTING: ++SSK Okmeydani Education Hospital, Eye Clinic, Turkiye Hospital, Okmeydani, Istanbul. METHODS: Three ablation zones were used in 92 eyes of 48 patients whose refractive errors were between -6.0 and -10.0 diopters (D) (mean spherical equivalent -7.42 D +/- 1.25 [SD]). The zones were between 4.5 and 6.0 mm based on the thickness of the cornea and the refractive correction. After the epithelium healed, dexamethasone was applied 4 times a day during the first postoperative week and then fluorometholone was applied 4 times a day for a minimum of 4 weeks. If hyperopia was found post-PRK, the steroid dose was gradually tapered. The patients were examined 1 and 3 days postoperatively, 1, 2, and 4 weeks, every 3 months for the first year, and then every 6 months. RESULTS: All patients were overcorrected in the first postoperative week. At 2 and 3 weeks, the mean manifest refraction was closer to emmetropia. At 6 months, the refraction was stable. The mean spherical equivalent was -0.10 D at the end of the first year, and stabilization continued for 4 years. After the third month, the haze regressed gradually without requiring treatment. In 1 patient, herpes simplex keratitis developed and healed in a short time with topical antiviral therapy. Nineteen eyes regressed more than -1.0 D, 4 eyes were overcorrected, 4 eyes had central islands (at 6 months), and 2 eyes were undercorrected. Two eyes were retreated for regression; 1 eye was retreated for undercorrection and 1 eye, for central island. An uncorrected visual acuity of 20/40 or better was achieved in 79.2% of eyes, and 73.9% were within +/-1. 0 D of the intended correction. CONCLUSION: ++Photorefractive keratectomy was effective in treating high myopia between -6.0 and -10.0 D. The induced refractive changes stabilized between 6 and 9 months. In most patients, no significant regression was found after this period. PMID- 11033396 TI - Choroidal neovascularization in myopic eyes after photorefractive keratectomy. AB - PURPOSE: To evaluate the incidence, characteristics, and results of treatment of choroidal neovascularization (CNV) in myopic eyes corrected by photorefractive keratectomy (PRK). SETTING: University Miguel Hernandez, Instituto Oftalmologico de Alicante, Alicante, Spain. METHODS: This study was of the incidence of CNV in 5936 consecutive eyes that had PRK for the correction of myopia. Mean follow-up was 38.5 months +/- 17.4 (SD). RESULTS: Extrafoveal CNV developed in the right eye of a 44-year-old woman 26 months after PRK for the correction of -12.00 diopters (D) of myopia. The follow-up after PRK was 38 months. Best corrected visual acuity (BCVA) before PRK was 20/40 (spherical equivalent [SE] -12.00 D). After PRK, BCVA was 20/32 SE -1.75 D). The CNV was treated by direct argon-green laser photocoagulation and did not recur in the subsequent 12 months). After CNV treatment, BCVA was 20/32 (SE -2.25 D). CONCLUSION: The incidence of CNV after PRK for myopia was low. Choroidal neovascularization is a possible complication in myopic eyes, and the risk exists before PRK. After PRK, the risk of CNV in myopic patients did not increase. PMID- 11033397 TI - Digital overlay technique for documenting toric intraocular lens axis orientation. AB - PURPOSE: To describe a digital overlay technique for documenting toric intraocular lens (IOL) axis alignment. SETTING: Jules Stein Eye Institute and the Department of Ophthalmology, University of California at Los Angeles School of Medicine, Los Angeles, California, USA. METHODS: Digital overlay imaging was used to evaluate the alignment of Staar toric IOLs in 4 eyes of 3 patients who had regular corneal astigmatism at the time of cataract surgery. Lens axes were determined by computerized analysis of digitally scanned retroillumination photographs. A stock digital image of a Staar AA4203TF toric IOL was superimposed on corneal topography images to document IOL alignment with the steep corneal meridian. RESULTS: Digital overlay images demonstrated that 3 IOLs in the sample group were within 5 degrees and 1 was within 20 degrees of the intended axis at the time of the final postoperative examination. Slitlamp lens axis estimations were 3 to 18 degrees different from that determined by this technique. CONCLUSION: Digital overlay imaging of correctly oriented toric IOLs on computerized corneal topography maps represents an intuitive, accurate, and visually appealing method of documenting toric IOL axis alignment. PMID- 11033398 TI - Corneal thickness and visual acuity after phacoemulsification with 3 viscoelastic materials. AB - PURPOSE: To determine whether differences exist in visual acuity and corneal thickness after phacoemulsification using various viscoelastic substances. SETTING: Phillips Eye Institute, Minneapolis, Minnesota, USA. METHODS: Fifty patients having routine phacoemulsification cataract extraction and intraocular lens implantation by a single surgeon using the same technique were randomly assigned to receive intraoperative administration of 1 of 3 viscoelastic substances: Amvisc Plus(R) (sodium hyaluronate 1.6%), OcuCoat(R) (hydroxypropyl methylcellulose 2%), or Viscoat(R) (chondroitin sulfate 4%-sodium hyaluronate 3%). Visual acuity and corneal thickness on the first postoperative day were compared between groups. RESULTS: Patients with a best corrected visual acuity (BCVA) of 20/40 or better 1 day postoperatively had significantly thinner corneas (596 microm) than those with a BCVA worse than 20/40 (639 microm). There were no differences in postoperative BCVA or percentage increase in central corneal pachymetry among the 3 viscoelastic groups. CONCLUSION: Viscoat, Amvisc Plus, and OcuCoat were comparable in their ability to produce clear corneas and good vision after routine phacoemulsification. PMID- 11033399 TI - Fourier analysis of irregular astigmatism after implantation of 3 types of intraocular lenses. AB - PURPOSE: To evaluate irregular astigmatism after silicone, acrylic, and poly(methyl methacrylate) (PMMA) intraocular lens (IOL) implantation using Fourier analysis of videokeratography data. METHODS: Two hundred forty eyes having phacoemulsification and IOL implantation were randomly assigned to 1 of 3 groups: 3.5 mm incision and silicone IOL, 4.1 mm incision and acrylic IOL, or 6.5 mm incision and PMMA IOL. All eyes had videokeratographic examinations preoperatively and 2, 4, and 10 days and 1 and 3 months postoperatively. The dioptric data of the central cornea were decomposed into spherical equivalent, regular astigmatism, and irregular astigmatism (decentration and higher-order irregularity) components using Fourier analysis. RESULTS: Regular astigmatism in the PMMA group was greater than in the silicone and acrylic groups. Decentration in all 3 groups increased significantly postoperatively but virtually returned to preoperative levels by 10 days. No significant difference was observed among the 3 groups. Higher-order irregularity significantly increased after surgery in all 3 groups. The higher-order irregularity in the PMMA group persisted for up to 1 month, while that in the silicone and acrylic groups returned to preoperative levels by 4 days, resulting in significant differences between these groups 10 days and 1 month after IOL implantation. CONCLUSION: Irregular astigmatism, both the decentration and higher-order irregularity components, increased significantly after 3 types of scleral tunnel incisions for silicone, acrylic, or PMMA IOL implantation but returned to preoperative levels soon after surgery except for the higher order irregularity after PMMA IOL implantation. PMID- 11033400 TI - Foldable silicone versus poly(methyl methacrylate) intraocular lenses in combined phacoemulsification and trabeculectomy. AB - PURPOSE: To compare the outcome of phacotrabeculectomy with implantation of poly(methyl methacrylate) (PMMA) or foldable silicone intraocular lenses (IOLs). METHODS: Thirty patients were randomized to receive a 5.5 mm PMMA IOL through a 5.0 mm incision or a foldable silicone IOL (Allergan SI-30) through a 3.2 mm incision. Visual acuity, intraocular pressure (IOP), bleb survival, inflammation, endothelial cell changes, and complications were examined at intervals up to 6 months. RESULTS: There was no difference between the 2 groups in final visual outcome, final IOP control, bleb survival, and endothelial cell changes. Two months after surgery, there was significantly more inflammation in the silicone IOL group than in the PMMA group (P <. 05). The silicone group had a significantly higher combined complication rate including iris capture, choroidal effusion, and epiretinal membrane formation (P <.05). CONCLUSIONS: Foldable silicone IOLs were comparable to conventional PMMA lenses in visual outcome, IOP control, bleb formation, and endothelial changes. However, some silicone lenses are associated with an increased risk of recurrence of inflammation and a higher final complication rate in combined cataract and filtration surgery. PMID- 11033401 TI - Manual multi-phacofragmentation through a 3.2 mm clear corneal incision. AB - PURPOSE: To evaluate the postoperative astigmatism and complications after manual multi-phacofragmentation (MPF) cataract surgery through a 3.2 mm clear corneal incision (CCI). SETTING: Department of Ophthalmology, Hospital General de Segovia, Segovia, Spain. METHODS: In a retrospective study, 50 eyes of 50 patients had cataract surgery by MPF. Patients were divided into 3 groups according to type of cataract. This technique uses a racquet-shaped nucleotome, a spatula, and 2 manipulators, right and left, for nuclear fragments. With these instruments, the nucleus is fragmented into multiple small pieces that are extracted from the anterior chamber. Astigmatism was evaluated before surgery and 7 days and 1 and 3 months after surgery by Gaussian distribution (P <.05). Intraoperative and postoperative complications were recorded. RESULTS: Mean postoperative astigmatism was 0.77 diopters (D) +/- 0. 29 (SD) at 7 days, 0.47 +/ 0.22 D at 1 month, and 0.21 +/- 0.22 D at 3 months. During surgery, 2 cases (4%) of slight transient intracameral bleeding in the anterior chamber occurred. After surgery, 5 eyes (10%) had corneal edema, 2 (4%) had iritis, and 3 (6%) had ocular hypertension. CONCLUSION: Using MPF, soft and hard nuclei can be removed through a 3.2 mm CCI. The astigmatic results were favorable and the complications few. PMID- 11033402 TI - Visual-evoked potentials: assessment of retrobulbar and peribulbar anesthesia. AB - PURPOSE: To assess the effects of retrobulbar and peribulbar anesthesia on nerve function as detected by visual-evoked potentials (VEPs). SETTING: University hospital in southern Brazil. METHODS: In a prospective study, 7 patients had peribulbar anesthesia and 9 had retrobulbar anesthesia for extracapsular cataract extraction. Visual-evoked potentials with pattern reversal and flash stimulation were performed at least 1 month before and 1 month after surgery. Study participants did not have ocular pathology other than cataract. The Lens Classification System III was used to grade the opacities before surgery. RESULTS: No significant difference was found between preoperative and postoperative evaluations in VEP flash and pattern-reversal amplitude and latency in either group (P >.05). Postoperative amplitude and latency was not significantly different between the peribulbar and retrobulbar groups. Two cases in the peribulbar group had altered wave morphology without clinical manifestation postoperatively. All patients had a final best spectacle-corrected visual acuity of 20/20. CONCLUSION: Block anesthetic procedures were safely used in cataract surgery, with no clinical sequelae to the optic nerve. PMID- 11033403 TI - Secondary glaucoma associated with crystalline lens subluxation. AB - PURPOSE: To elucidate the clinical characteristics of secondary glaucoma associated with subluxation of the crystalline lens. SETTING: Department of Ophthalmology and Visual Sciences, Kyoto University Graduate School of Medicine, Kyoto, and Department of Ophthalmology, Tenri Hospital, Nara, Japan. METHODS: This retrospective study comprised 14 eyes of 13 patients with uncontrolled intraocular pressure (IOP) and lens subluxation. The subluxated lens was extracted through surgery. RESULTS: Angle closure caused by the subluxated lens was complicated in 3 eyes. In the remaining 11 eyes, uncontrolled IOP elevation was found despite the presence of deep anterior chambers and wide open angles. A mean of 14.1 months +/- 13.7 (SD) after cataract surgery, IOP was well controlled (lower than 21 mm Hg) in all 14 eyes. Mean IOP was 15.4 +/- 2.2 mm Hg at the final examination. Complications included transient vitreous hemorrhage in 5 eyes, choroidal detachment in 2 eyes, and retinal tears in 1 eye. CONCLUSION: Lens extraction surgery was effective in controlling IOP in eyes with secondary glaucoma associated with lens subluxation. PMID- 11033404 TI - Effect of mitomycin-C on posterior capsule opacification in rabbit eyes. AB - PURPOSE: To determine whether mitomycin-C can inhibit posterior capsule opacification (PCO) without causing ocular toxicity. SETTING: Yonsei Institute of Vision Research, Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Korea. METHODS: Mitomycin-C dissolved in sodium hyaluronate (0.2 cc of 0.2 mg/mL) was injected into the empty capsular bag for 3 minutes after endocapsular phacoemulsification in rabbit eyes. Three months after surgery, the obstruction rate of visible light caused by PCO was measured using an optical power meter. RESULTS: The mean obstruction rate of visible light was 81.0% +/- 8.3% (SD) in the control group in which sodium hyaluronate without mitomycin-C was used, 30.5% +/- 10.1% in the group in which mitomycin-C was dissolved in sodium hyaluronate, and 71.9% +/- 6.8% in the group in which mitomycin-C was dissolved in a balanced salt solution. Statistically significant differences were found among all 3 groups. CONCLUSIONS: Our results suggest that the application of mitomycin-C dissolved in sodium hyaluronate effectively reduces PCO in rabbit eyes. PMID- 11033405 TI - Preventing lens epithelial cell migration using intraocular lenses with sharp rectangular edges. AB - PURPOSE: To compare the preventive effect on posterior capsule opacification (PCO) of an acrylic intraocular lens (IOL) and a silicone IOL, both with sharp rectangular optic edges. SETTING: Jinshikai Medical Foundation, Nishi Eye Hospital, Osaka, Japan. METHODS: After phacoemulsification, an acrylic IOL (AcrySof(R)) was implanted in 1 eye and a silicone IOL (CeeOn 911) in the contralateral eye of 7 rabbits, one of which was omitted from evaluation. RESULTS: The Miyake-Apple view and histopathological findings 3 weeks after surgery revealed that the lens capsule wrapped tightly around the optic edges, conforming to a distinctly sharp bend, with both IOL types in 5 rabbits. Migrating lens epithelial cells (LECs) were inhibited at the site. The AcrySof IOL had a better capsular bend effect in the histological sections, but there was no apparent difference in PCO development between the 2 IOLs. CONCLUSIONS: The CeeOn 911 silicone IOL, with sharp rectangular optic edges, showed an effect on preventing PCO that was similar to that of the AcrySof IOL. When a discontinuous capsular bend is created by an IOL with sharp optic edges, the bend may induce contact inhibition of migrating LECs regardless of the lens' material composition. PMID- 11033406 TI - Effect of surgical technique on in vitro posterior capsule opacification. AB - PURPOSE: To compare the effect of different cataract extraction surgical techniques on residual lens epithelial cell (LEC) density and cell regrowth rates using an in vitro model of posterior capsule opacification (PCO). SETTING: Comparative Ophthalmology Research Laboratories, North Carolina State University, Raleigh, North Carolina, USA. METHODS: Lens capsule explants were prepared from freshly enucleated canine globes after extracapsular cataract extraction (ECCE), phacoemulsification, or phacoemulsification followed by capsule vacuuming. Initial cell density on the capsule and cell proliferation were determined by phase contrast microscopy. The effects of the surgical technique on time to confluent growth of the cells across the posterior lens capsule were determined. RESULTS: Residual cell density on the remaining anterior capsule immediately after lens removal was 31.6% +/- 19.3%, 16.1% +/- 8.9%, and 7.7% +/- 5.7% in the ECCE, phacoemulsification, and phacoemulsification/capsule-vacuuming groups, respectively. Time to confluence (range 5.0 to 6.3 days) was not significantly different among the 3 groups when the lens capsules were cultured in serum supplemented media. The confluence rate was significantly longer (by approximately 5 to 7 days) in the phacoemulsification/capsule-vacuuming group than in the other 2 groups when the capsules were cultured in serum-free media. CONCLUSIONS: Phacoemulsification with and without anterior and equatorial capsular vacuuming led to less initial LEC density in the capsular bag than ECCE. However, because cell proliferation rates among the 3 groups were only marginally affected, near 100% removal of LEC at the time of cataract extraction may be necessary to prevent PCO. PMID- 11033407 TI - Persisting diplopia after bilateral laser in situ keratomileusis. AB - We present a case of persisting diplopia after bilateral laser in situ keratomileusis in a patient with high anisometropia and amblyopia. Treatment of this complication by ocular-muscle surgery was not possible because of missing fusion. We suggest that, especially in cases with anisometropia, a complete examination of binocularity be performed before refractive surgery. PMID- 11033408 TI - Posterior capsule tear with plate-haptic silicone intraocular lens dislocation. AB - We report a case of posterior capsule rupture and posterior dislocation of an intraocular lens (IOL) caused by the tip of a silicone plate-haptic lens. The dislocation occurred during IOL implantation after uneventful phacoemulsification in a patient with a small pupil. Discussed are the possible reasons for the complication and measures to prevent it. PMID- 11033409 TI - Pit defects on the anterior surface of hydrophilic foldable intraocular lenses. AB - A series of pits varying in depth and size appeared on the anterior surface of hydrogel intraocular lenses (IOLs) soon after they were implanted. In all cases, the pits increased in size but not in number during a 1 year follow-up. The origin of the pit defects is unknown and does not seem related to inflammatory or lens epithelial cell activity. Hydration of the IOL could be a cause even if a manufacturing defect were present. This IOL surface alteration did not seem to impair visual function or cause visual disturbances. PMID- 11033410 TI - Overexpression of CuZnSOD in coronary vascular cells attenuates myocardial ischemia/reperfusion injury. AB - Superoxide dismutase scavenges oxygen radicals, which have been implicated in ischemia/reperfusion (I/R) injury in the heart. Our experiments were designed to study the effect of a moderate increase of copper/zinc superoxide dismutase (CuZnSOD) on myocardial I/R injury in TgN(SOD1)3Cje transgenic mice. A species of 0.8 kb human CuZnSOD mRNA was expressed, and a 273% increase in CuZnSOD activity was detected in the hearts of transgenic mice with no changes in the activities of other antioxidant enzymes. Furthermore, immunoblot analysis revealed no changes in the levels of HSP-70 or HSP-25 levels. Immunocytochemical study indicated that there was increased labeling of CuZnSOD in the cytosolic fractions of both endothelial cells and smooth muscle cells, but not in the myocytes of the hearts from transgenic mice. When these hearts were perfused as Langendorff preparations for 45 min after 35 min of global ischemia, the functional recovery of the hearts, expressed as heart rate x LVDP, was 48 +/- 3% in the transgenic hearts as compared to 30 +/- 5% in the nontransgenic hearts (p <.05). The improved cardiac function was accompanied by a significant reduction in lactate dehydrogenase release from the transgenic hearts. Our results demonstrate that overexpression of CuZnSOD in coronary vascular cells renders the heart more resistant to I/R injury. PMID- 11033411 TI - The level of typical biomarker of oxidative stress 8-hydroxy-2'-deoxyguanosine is higher in uterine myomas than in control tissues and correlates with the size of the tumor. AB - The results of this work show a higher level of 8-hydroxy-2'-deoxyguanosine (8-OH dG), a typical biomarker of oxidative stress, in uterine myoma tissues than in their respective tumor-free tissues. The level of this modified base was elevated in uterine tissues of premenopausal women when compared with postmenopausal ones. We have also found the correlation between the size of the tumor and the amount of 8-OH-dG. These results suggest that estrogen-produced 8-OH-dG may be one of the factors responsible for the formation of the myoma, and it may contribute to malignant transformation of myoma cells. PMID- 11033412 TI - Garlic ameliorates gentamicin nephrotoxicity: relation to antioxidant enzymes. AB - Reactive oxygen species are involved in gentamicin (GM) nephrotoxicity, and garlic is effective in preventing or ameliorating oxidative stress. Therefore, the effect of garlic on GM nephrotoxicity was investigated in this work. Four groups of rats were studied: (i) fed normal diet (CT), (ii) treated with GM (GM), (iii) fed 2% garlic diet (GA), and (iv) treated with GM and 2% garlic diet (GM + GA). Rats were placed in metabolic cages and GM nephrotoxicity was induced by injections of GM (75 mg/kg every 12 h) for 6 d. Lipoperoxidation and enzyme determinations were made in renal cortex on day 7. GM nephrotoxicity was made evident on day 7 by (i) tubular histological damage, (ii) enhanced BUN and urinary excretion of N-acetyl-beta-D-glucosaminidase, and (iii) decreased creatinine clearance. These alterations were prevented or ameliorated in GM + GA group. The rise in lipoperoxidation and the decrease in Mn-SOD and glutathione peroxidase (GPx) activities observed in the GM group, were prevented in the GM + GA group. Cu, Zn-SOD activity and Mn-SOD and Cu,Zn-SOD content did not change. CAT activity and content decreased in the GM, GA, and GM + GA groups. CAT mRNA levels decreased in the GM group. The protective effect of garlic is associated with the prevention of the decrease of Mn-SOD and GPx activities and with the rise of lipoperoxidation in renal cortex. PMID- 11033413 TI - Effects of oxidative stress on the expression of antioxidative defense enzymes in spontaneously hypertensive rat hearts. AB - Little is known concerning the effect of oxidative stress on the expression of antioxidative enzymes in the decompensated cardiac hypertrophy of spontaneously hypertensive rats (SHR), considered as a model of dilative cardiomyopathy in man. Superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) were characterized in isolated perfused hearts of 18 month old SHR and the age-matched normotensive control Wistar-Kyoto (WKY) rats, before and after 30 min infusion of 25 microM H(2)O(2). After infusion of H(2)O(2), aortic flow decreased in WKY from 26.2 +/- 2.2 to 16.0 +/- 0.8 ml/min (p <.05) but not in SHR (18.2 +/- 1.9 vs. 20.7 +/- 2.2 ml/min). This protection was related to the higher myocardial activities of GPx, MnSOD and CuZnSOD in SHR, compared with those of the WKY group. Although total SOD activity in the SHR fell after H(2)O(2) exposure (to 1.81 +/- 0.13 from 3.56 +/- 0.49 U/mg of protein), catalase activity increased (to 2.46 +/- 0.34 from 1.56 +/- 0.29 k min(-1)mg(-1)protein), compared with the pre-infusion period (p <.05 in each case). In additional studies, hearts were subjected to 30 min of global ischemia followed by 30 min of reperfusion. The results obtained in ischemic/reperfused hearts show the same changes in enzyme activities measured as it was observed in H(2)O(2) perfused hearts, indicating that oxidative stress is independent of the way it was induced. The higher catalase activity derived from elevated mRNA synthesis. The antioxidative system in dilative cardiomyopathic hearts of SHR is induced, probably due to episodes of oxidative stress, during the process of decompensation. This conditioning of the antioxidative potential may help overcome acute stress situations caused by reactive oxygen species in the failing myocardium. PMID- 11033414 TI - Peroxidation of arachidonate containing plasmenyl glycerophosphocholine: facile oxidation of esterified arachidonate at carbon-5. AB - Oxidation of 1-O-hexadec-1'-enyl-arachidonoyl glycerophosphocholine (16:0p/20:4 GPC) by hydroxyl radical generated from Cu(II)/H(2)O(2) was found to yield major products corresponding to free carboxylic acids of 5-hydroxyeicosatetraenoic acid and several 5, 12-dihydroxyeicosatetraenoic acid. These products were characterized by electrospray tandem mass spectrometry based upon characteristic product ion spectra, as well as HPLC retention time. Several products were found to be biologically active in terms of elevating neutrophil intracellular calcium ion concentration. When mixed micelles of 16:0p/20:4-GPC were treated with Cu(II)/H(2)O(2), oxidation of the arachidonate esterified to the plasmalogen glycerophosphocholine lipid resulted in the most abundant products oxidized at carbon-5 of esterified arachidonate, but free carboxylic acid products were not formed. The mechanism of formation of these oxidized products is suggested to involve a cooperation between the sn-1 vinyl ether substituent and the arachidonoyl substituent at sn-2 of the glycerophospholipid to direct oxidation of the arachidonate ester at carbon-5. Since arachidonic acid is found in high abundance within most plasmalogen glycerophospholipids, the susceptibility of plasmalogens to free radical oxidation likely involves concomitant oxidation of the arachidonyl radyl group esterified at the sn-2 position. PMID- 11033415 TI - The nitroxide tempol induces oxidative stress, p21(WAF1/CIP1), and cell death in HL60 cells. AB - The antiproliferative effect of Tempol, a stable nitroxide free radical, was investigated on the p53-negative human leukemia cell line HL60. A concentration- and time-dependent inhibition of cell growth was observed that appears to be due to induction of apoptosis. Involvement of oxidative stress is indicated by a concentration-dependent increase in intracellular peroxides and a parallel decrease in total cellular glutathione; in addition, increased survival rates were observed in cells simultaneously treated with Tempol and the antioxidant N acetylcysteine. Tempol did not affect the relative levels of Bax and Bcl2, whereas p21(WAF1/CIP1) was enhanced in a concentration- and time-dependent fashion; this effect was partially inhibited by N-acetylcysteine, was maintained for up to 8 h after Tempol removal, and seemed to depend on continuing protein synthesis. The increase in p21(WAF1/CIP1) was accompanied by a parallel accumulation of cells in the G(1) phase of the cycle and by a decrease in the 110 kDa form of pRb. Our results suggest that p53-independent induction of p21(WAF1/CIP1) mediates the antiproliferative effect of Tempol; on the basis of this observation, the nitroxide could be proposed as an useful adjunct to the treatment of p53-deficient tumors, which are often refractory to standard chemotherapy. PMID- 11033416 TI - Metabolism of lipid peroxidation product, 4-hydroxynonenal (HNE) in rat erythrocytes: role of aldose reductase. AB - Lipid peroxidation represents a significant source of erythrocyte dysfunction and aging. Because the toxicity of lipid peroxidation appears to be in part due to aldehydic end products, we examined, in rat erythrocytes, the metabolism of 4 hydroxy-trans-2-nonenal (HNE), one of the most abundant and toxic lipid-derived aldehydes. Packed erythrocytes, 0.1 ml, completely metabolized 20 nmoles of HNE in 20 min. The glutathione conjugate of HNE and 4-hydroxynonanoic acid (HNA) represented 70 and 25% of the total metabolism, respectively. Approximately 70% of the metabolites were extruded to the medium. Upon electrospray ionization mass spectrometry, the glutathione conjugate resolved into two distinct species corresponding to glutathionyl HNE (GS-HNE) and glutathionyl 1,4-dihydroxynonene (GS-DHN). The concentration of GS-DHN formed was twice that of GS-HNE. Inhibition of aldose reductase by sorbinil and tolrestat led to a selective decrease in the formation of GS-DHN, although the extent of HNE glutathiolation was unaffected. Inhibitors of aldehyde or alcohol dehydrogenase, i.e., cyanamide and 4-methyl pyrazole, had no effect on the formation of HNA and GS-DHN, indicating that these enzymes are not significant participants in the erythrocyte HNE metabolism. Thus, oxidation to HNA, conjugation with glutathione, and further reduction of the conjugate by aldose reductase appear to be the major pathways of HNE metabolism in erythrocytes. These pathways may be critical determinants of erythrocyte toxicity due to lipid peroxidation-derived aldehydes. PMID- 11033417 TI - Increased oxidative damage to DNA in ALS patients. AB - Although the cause of amyotrophic lateral sclerosis (ALS) is unknown, substantial evidence indicates that oxidative toxicity is associated with neuronal death in this disease. We examined levels of a well-established marker of oxidative damage to DNA, 8-hydroxy-2'-deoxyguanosine (8OH2'dG) in plasma, urine, and cerebrospinal fluid (CSF) at a single time point from subjects with ALS, other neurological diseases, or no known disorders. We also measured the rate of change of 8OH2'dG levels in plasma and urine from ALS and in urine from control subjects over 9 months and examined the relationship to disease severity. In each fluid, 8OH2'dG levels were significantly elevated in the ALS group as compared to control subjects. In all subjects, the plasma and CSF 8OH2'dG levels increased with age, providing further evidence for a role of oxidative damage in normal aging. Plasma and urine 8OH2'dG levels increased significantly with time in the ALS group only. The rate of increase in urine 8OH2'dG levels with time was significantly correlated with disease severity. These findings are consistent with the hypothesis that oxidative pathology accompanies the neurodegenerative process in ALS and suggest that 8OH2'dG may provide a useful tool for monitoring therapeutic interventions in this disease. PMID- 11033418 TI - The origin of red cell fluorescence caused by hydrogen peroxide treatment. AB - Fluorescence in red cells following hydrogen peroxide treatment has been attributed to lipid peroxidation of the membrane. The putative relationship between lipid peroxidation and fluorescence was questioned by the finding that BHT and alpha-tocopherol, which are thought to inhibit lipid peroxidation, do not inhibit the fluorescence detected by flow cytometry. Furthermore, lipid peroxidation induced in red cells by the Fe(III)-ADP-ascorbate system did not produce fluorescence. These results require an alternative explanation for the hydrogen peroxide-induced fluorescence. A role for reduced hemoglobin is indicated by the inhibition of fluorescence by pretreatment of cells with CO that binds strongly to ferrohemoglobin and nitrite that oxidizes ferrohemoglobin. Our earlier studies have shown the formation of fluorescent heme degradation products during the reaction of purified hemoglobin with hydrogen peroxide, which was also inhibited by CO and nitrite pretreatment. The fluorescence produced in red cells after the addition of hydrogen peroxide can, therefore, be attributed to fluorescent heme degradation products. PMID- 11033419 TI - Levels of DNA damage are unaltered in mice overexpressing human catalase in nuclei. AB - Two types of transgenic mice were generated to evaluate the role of hydrogen peroxide in the formation of nuclear DNA damage. One set of lines overexpresses wild-type human catalase cDNA, which is localized to peroxisomes. The other set overexpresses a human catalase construct that is targeted to the nucleus. Expression of the wild-type human catalase transgene was found in liver, kidney, skeletal muscle, heart, spleen, and brain with muscle and heart exhibiting the highest levels. Animals containing the nuclear-targeted construct had a similar pattern of expression with the highest levels in muscle and heart, but with lower levels in liver and spleen. In these animals, immunofluorescence detected catalase present in the nuclei of kidney, muscle, heart, and brain. Both types of transgenic animals had significant increases of catalase activities compared to littermate controls in most tissues examined. Despite enhanced activities of catalase, and its presence in the nucleus, there were no changes in levels of 8OHdG, a marker of oxidative damage to DNA. Nor were there differences in mutant frequencies at a Lac Z reporter transgene. This result suggests that in vivo levels of H(2)O(2) may not generate 8OHdG or other types of DNA damage. Alternatively, antioxidant defenses may be optimized such that additional catalase is unable to further protect nuclear DNA against oxidative damage. PMID- 11033420 TI - Suppression of NF-kappaB activation and cytokine production by N-acetylcysteine in pancreatic acinar cells. AB - Reactive oxygen species (ROS), generated by infiltrating neutrophils, are considered as an important regulator in the pathogenesis and development of pancreatitis. A hallmark of the inflammatory response is the induction of cytokine gene expression, which may be regulated by oxidant-sensitive transcription factor, nuclear factor-kappaB (NF-kappaB). Present study aims to investigate whether neutrophils primed by 4beta-phorbol 12beta-myristate 13alpha acetate (PMA) affect the productions of H(2)O(2) and lipid peroxide (LPO), NF kappaB activation and cytokine production in pancreatic acinar cells, and whether these alterations were inhibited by N-acetylcysteine (NAC) and superoxide dismutase (SOD). Neutrophils generated ROS by stimulation with PMA, which was inhibited by NAC and SOD. In acinar cells, PMA-primed neutrophils increased the productions of H(2)O(2), LPO, and cytokines both time and dose dependently. PMA primed neutrophils resulted in the activation of two species of NF-kappaB dimers (a p50/p65 heterodimer and a p50 homodimer) in acinar cells. Both NAC and SOD inhibited neutrophil-induced, oxidant-mediated alterations in acinar cells. In conclusion, ROS, generated by neutrophils, activates NF-kappaB, resulting in upregulation of inflammatory cytokines in acinar cells. Antioxidants such as NAC might be useful antiinflammatory agents by inhibiting oxidant-mediated activation of NF-kappaB and decreasing cytokine production. PMID- 11033422 TI - Frontiers in biopharmacy PMID- 11033421 TI - Hydrogen peroxide inhibits activation, not activity, of cellular caspase-3 in vivo. AB - Oxidants such as H(2)O(2) can induce a low level of apoptosis at low concentrations but at higher concentrations cause necrosis. Higher concentrations of H(2)O(2) also inhibit the induction of apoptosis by chemotherapy drugs. One theory is that, at higher concentrations, H(2)O(2) causes direct oxidative inactivation of caspase-3 activity, thus preventing the apoptotic pathway from being used. We find that treatment of recombinant caspase-3 with H(2)O(2) can partially reduce its enzymatic activity: However, the following findings show that this does not occur in the cell. (1) The inhibition by H(2)O(2) of VP-16 induced apoptosis and cellular caspase-3 activity can be overcome by adding inhibitors of poly(ADP-ribose) polymerase (PARP) at sub-stoichiometric concentrations. (2) Delayed addition of H(2)O(2) to VP-16-treated cells prevents additional caspase induction but does not inhibit the caspase activity that has already been generated. (3) H(2)O(2) is a poor inhibitor of caspase-3 activity in cell lysates. (4) Addition of H(2)O(2) to cells inhibits activation of caspase-9, which is required for activation of caspase-3. We conclude that inhibition of caspase-3 activity in the cell occurs indirectly at a step located upstream of caspase-3 activation. H(2)O(2) acts in part by inducing DNA strand breaks and activating PARP, thus depleting the cells of ATP. When this pathway is blocked, even high concentrations of H(2)O(2) can induce caspase-9 and -3 activation and cause apoptosis. PMID- 11033423 TI - Molecules as complex adaptative systems: constrained molecular properties and their biochemical significance. AB - The first part of the paper describes molecular structure by considering form (geometrical structure), function (observable properties resulting from interaction with a probe) and fluctuation (dynamics). The fluctuation of form and function generates a number of molecular states, whose ensemble delineates a property space. This concept is central when examining the mutual interactions of a chemical compound with its molecular environment. Because of these interactions, a chemical compound and its molecular environment may form a complex system in its own right, which exhibits emergent properties (e.g., solubility and lipophilicity) that are non-existent (and meaningless) at the level of description of isolated molecules, and which are part of the property space of the compound. The emergence of these properties is accompanied by constraints on the property space of the constituents, and especially of the chemical compound. Some of these constraints are well known, e.g., conformational shifts as seen in induced fit and chameleonic behaviour. Such mutual adaptability between a compound and its environment is a fundamental but insufficiently recognized phenomenon in biochemistry, since it amplifies molecular complementarity and hence molecular recognition. In the second part of this paper, the results of an exploratory study are reported which show that contraints on the property space of constituents also occur when molecules are covalently incorporated into larger compounds. Using the GRID/VolSurf softwares, we bring evidence that when some amino acids become residues in peptides, they experience a modest increase in their polarity and a marked increase in their hydrophobicity (as assessed by their polarity field and hydrophobicity field, respectively). The biological and pharmacological implications of constraints on solutes, ligands and monomers could inspire new directions of research. PMID- 11033424 TI - Rationale and applications of lipids as prodrug carriers. AB - Lipidic prodrugs, also called drug-lipid conjugates, have the drug covalently bound to a lipid moiety, such as a fatty acid, a diglyceride or a phosphoglyceride. Drug-lipid conjugates have been prepared in order to take advantage of the metabolic pathways of lipid biochemistry, allowing organs to be targeted or delivery problems to be overcome. Endogenous proteins taking up fatty acids from the blood stream can be targeted to deliver the drug to the heart or liver. For glycerides, the major advantage is the modification of the pharmacokinetic behavior of the drug. In this case, one or two fatty acids of a triglyceride are replaced by a carboxylic drug. Lipid conjugates exhibit some physico-chemical and absorption characteristics similar to those of natural lipids. Non-steroidal, anti-inflammatory drugs such as acetylsalicylic acid, indomethacin, naproxen and ibuprofen were linked covalently to glycerides to reduce their ulcerogenicity. Mimicking the absorption process of dietary fats, lipid conjugates have also been used to target the lymphatic route (e.g., L-Dopa, melphalan, chlorambucil and GABA). Based on their lipophilicity and resemblance to lipids in biological membranes, lipid conjugates of phenytoin were prepared to increase intestinal absorption, whereas glycerides or modified glycerides of L Dopa, glycine, GABA, thiorphan and N-benzyloxycarbonylglycine were designed to promote brain penetration. In phospholipid conjugates, antiviral and antineoplasic nucleosides were attached to the phosphate moiety. After presenting the biochemical pathways of lipids, the review discusses the advantages and drawbacks of lipidic prodrugs, keeping in mind the potential pharmacological activity of the fatty acid itself. PMID- 11033425 TI - VolSurf: a new tool for the pharmacokinetic optimization of lead compounds. AB - A method for the modeling and prediction of pharmacokinetic properties based on computed molecular interaction fields and multivariate statistics has been investigated in different experimental datasets. The program VolSurf was used to correlate 3D molecular structures with physico-chemical and pharmacokinetic properties. In membrane partitioning, VolSurf produced a two-component model explaining 94% of the total variation with a predictive q(2) of 0.90. This result was achieved without conformational sampling and without any quantum-chemical calculation. For the prediction of blood-brain barrier penetration the VolSurf model was able to predict the BBB profile for most of the drugs in the external prediction set. In Caco-2 and MDCK permeation experiments, VolSurf was used with success to establish statistical models and to predict the behaviour of new compounds. The method thus appears as a valuable new property filter in virtual screening and as a novel tool in optimizing the pharmacokinetic profile of pharmaceutically relevant compounds. PMID- 11033426 TI - Membrane transporters. AB - Carrier-mediated drug transport is relatively unexplored in comparison with passive transcellular and paracellular drug transport. Yet, there is a host of transporter proteins that can be targeted for improving epithelial drug absorption. Generally, these are transport mechanisms for amino acids, dipeptides, monosaccharides, monocarboxylic acids, organic cations, phosphates, nucleosides, and water-soluble vitamins. Among them, the dipeptide transporter mechanism has received the most attention. Dipeptide transporters are H(+) coupled, energy-dependent transporters that are known to play an essential role in the oral absorption of beta-lactam antibiotics, angiotensin-converting enzyme (ACE) inhibitors, renin inhibitors, and an anti-tumor drug, bestatin. Moreover, several investigators have demonstrated the utility of the dipeptide transporter as a platform for improving the oral bioavailability of drugs such as zidovudine and acyclovir through dipeptide prodrug derivatization. Thus far, at least four proton-coupled peptide transporters have been cloned. The first one cloned was PepT1 from the rabbit small intestine. The focus of this presentation will be structure-function, intracellular trafficking, and regulation of PepT1. Disease, dietary, and possible excipient influences on PepT1 function will also be discussed. PMID- 11033427 TI - Cell cultures as tools in biopharmacy. AB - A survey is given on a few selected cell culture models that are used for transport studies. They are characterised for growth, transcellular electrical resistance and cytoarchitecture. The importance of standardisation in view of their use as transport models is documented. Their potential for studies on passive permeation and P-glycoprotein-mediated transport is explored and related to published data. Transport studies are presented that were performed in a two chamber set-up, the Costar "vertical diffusion system". A series of non homologous compounds showed similar permeability data (P(app)) in the different cell cultures. The origin of the cell type had no remarkable influence on passive transcellular permeation. MDCK cells, an epithelial cell line of canine kidney origin, are perfectly suited to screen for passive permeation. They have low expression of transporter proteins and low metabolic activity. In general, they probably represent the best-known epithelial cell line with respect to genetics as well as lipid and protein composition. MDCK cells are easy to handle. Transport experiments can be done between 7 and 14 days after seeding, when the stationary growth phase is reached. To screen for P-glycoprotein substrates, efflux and uptake studies were performed with mdr1-transfected MDCK cells (MDR1 MDCK) in a one-chamber system in the presence or absence of verapamil or cyclosporin A as inhibitor. Evidence is presented why the transfected cells, which express large amounts of P-glycoprotein, are not suitable for two-chamber transport studies. PMID- 11033428 TI - Rapid assessment of drug metabolism in the drug discovery process. AB - For a few years, in vitro models have been used as part of high-throughput screening (HTS) programs to characterize metabolic stability, drug permeability and drug solubility. This has allowed the rapid selection of lead candidates based not only on pharmacological endpoints but also on biopharmaceutical specifications. What has now become clear is that the huge amount of data produced to sort series of compounds has a limited predictive value when used to predict human pharmacokinetic parameters. More complex in vitro tests together with some simple in vivo tests used as validation steps have been developed in order to provide absolute data that may be used as a complement to lead selection providing reliable predictions not only of human pharmacokinetics but also of potential drug-drug interactions. These models may be used as part of selective drug screening (SDS) programs. Further advances in analytical and in vitro techniques will see some of these models shifting from SDS to HTS programs putting the emphasis on the use of expert systems and physiologically based pharmacokinetic models (PBPK) to provide meaningful endpoint data. PMID- 11033429 TI - In vitro-in vivo correlations for lipophilic, poorly water-soluble drugs. AB - Although several routes of administration can be considered for new drug entities, the most popular remains the oral route. To predict the in vivo performance of a drug after oral administration from in vivo data, it is essential that the limiting factor to absorption can be modelled in vitro. In the case of BCS class II drugs dissolution is rate-limiting to absorption, so the use of biorelevant dissolution tests can be used to predict differences in bioavailability among different formulations and dosing conditions. To achieve an a priori correlation, the composition, volume and hydrodynamics of the contents in the gastrointestinal lumen following administration of the dosage form must be accurately simulated. Four media have been chosen/developed to model composition of the gastric and intestinal contents before and after meal intake. These are SGF, milk, FASSIF and FeSSIF, which model fasted and fed state conditions in the stomach and small intestine respectively. Using these media, excellent correlations have been obtained with the following poorly soluble drugs: danazol, ketoconazole, atovaquone and troglitazone. In all cases, fed vs. fasted state effects can be predicted from dissolution data and, where several formulations were available for testing, dissolution tests could also be used to determine which would have the best in vivo performance. PMID- 11033430 TI - Drug targeting. AB - The main problems currently associated with systemic drug administration are: even biodistribution of pharmaceuticals throughout the body; the lack of drug specific affinity toward a pathological site; the necessity of a large total dose of a drug to achieve high local concentration; non-specific toxicity and other adverse side-effects due to high drug doses. Drug targeting, i.e. predominant drug accumulation in the target zone independently on the method and route of drug administration, may resolve many of these problems. Currently, the principal schemes of drug targeting include direct application of a drug into the affected zone, passive drug targeting (spontaneous drug accumulation in the areas with leaky vasculature, or Enhanced Permeability and Retention-EPR-effect), 'physical' targeting (based on abnormal pH value and/or temperature in the pathological zone), magnetic targeting (or targeting of a drug immobilized on paramagnetic materials under the action of an external magnetic field), and targeting using a specific 'vector' molecules (ligands having an increased affinity toward the area of interest). The last approach provides the widest opportunities. Such pharmaceutical carriers as soluble polymers, microcapsules, microparticles, cells, cell ghosts, liposomes, and micelles have been successfully used for targeted drug delivery in vivo. Though the direct conjugation of a drug molecule with a targeted moiety is also possible (immunotoxin), the use of microreservoir type systems provides clear advantages, such as high loading capacity, possibility to control size and permeability of drug carrier systems and use relatively small number of vector molecules to deliver substantial quantities of a drug to the target. The practical use of the listed systems and approaches for the delivery of therapeutic and diagnostic agents will be considered. PMID- 11033431 TI - Lipid formulations for oral administration of drugs: non-emulsifying, self emulsifying and 'self-microemulsifying' drug delivery systems. AB - 'Lipid' formulations for oral administration of drugs generally consist of a drug dissolved in a blend of two or more excipients, which may be triglyceride oils, partial glycerides, surfactants or co-surfactants. The primary mechanism of action which leads to improved bioavailability is usually avoidance, or partial avoidance, of the slow dissolution process which limits the bioavailability of hydrophobic drugs from solid dosage forms. Ideally the formulation allows the drug to remain in a dissolved state throughout its transit through the gastrointestinal tract. The availability of the drug for absorption can be enhanced by presentation of the drug as a solubilizate within a colloidal dispersion. This objective can be achieved by formulation of the drug in a self emulsifying system or alternatively by taking advantage of the natural process of triglyceride digestion. In practice 'lipid' formulations range from pure oils, at one extreme, to blends which contain a substantial proportion of hydrophilic surfactants or cosolvents. Knowledge of the efficiency of emulsification of these formulations, the nature of the colloidal system formed by dispersion, their susceptibility to digestion, and the subsequent fate of the drug is desirable for formulation. Yet the literature on this subject is limited, so this article represents part review and part commentary on current status of lipid formulations. A simple classification system for lipid formulations, based on the polarity of the blend and reviewed here, will help comparison of data between laboratories. Priorities for future work are discussed. More data is needed on the solubility of drugs in various types of formulations, and in particular, on the relationship between the physical chemistry of the drug and its fate, subsequent to dilution and digestion of the formulation in the lumen of the gastrointestinal tract. The mechanisms of action and practical uses of each type of lipid formulation are discussed. PMID- 11033432 TI - New methods for screening and drug characterization. Foreword. PMID- 11033433 TI - The measurement of efficacy in the drug discovery agonist selection process. PMID- 11033434 TI - Use of a cell-based, lawn format assay to rapidly screen a 442,368 bead-based peptide library. AB - A cell-based, lawn format assay utilizing an in situ photocleavage method has been developed that allows the rapid examination of large bead-based compound libraries as discrete molecules. The format uses frog melanophore cells in a contiguous, adherent, confluent layer in small petri dishes covered with a 0.5-1 mm layer of agarose containing 130 micron diameter TentaGel beads at a density of 2-20 beads/mm2. Employing this technique a 9-mer, 442,368-member peptide library (designed around the 13 amino acid alpha-MSH peptide sequence) made up of 12 separate pools of 36,864 peptides/pool was assayed. Initially, a fraction (approximately 10%) of each pool was scanned (approximately 3700 beads from each pool) in 60-mm petri dishes to identify the most active pools. Upon direct photocleavage of the beads with UV light (365 nm), each petri dish was photographed over a 60-min period with a CCD camera to record changes in light intensity as an index of melanosome dispersion. Active beads were those that were surrounded by a localized decrease in light transmittance indicating melanosome dispersed cells. Upon examination with a dissecting microscope, single beads centrally located to a circular array of dispersed cells were identified and removed from the agarose and sequenced by Edman degradation to determine the peptide sequence. Re-synthesized peptides were re-examined against alpha-MSH receptor to confirm and quantify the activity. Several 9-mer peptides were identified with potencies similar to the natural 13-mer peptide. This method allows for the rapid screening of large bead-based photo-cleavable peptide libraries with the advantage that each compound is screened as a discrete molecule in a well-less format. PMID- 11033435 TI - Constitutive receptor systems for drug discovery. AB - This paper discusses the use of constitutively active G-protein-coupled receptor systems for drug discovery. Specifically, the ternary complex model is used to define the two major theoretical advantages of constitutive receptor screening namely, the ability to detect antagonists as well as agonists directly and the fact that constitutive systems are more sensitive to agonists. In experimental studies, transient transfection of Chinese hamster ovary cyclic AMP response element (CRE) luciferase reporter cells with cDNA for human parathyroid hormone receptor, glucagon receptor, and glucagon-like peptide (GLP-1) receptor showed cDNA concentration-dependent constitutive activity with parathyroid hormone (PTH 1) and glucagon. In contrast, no constitutive activity was observed for GLP-1 receptor, yet responses to GLP-1 indicated that receptor expression had taken place. In another functional system, Xenopus laevi melanophores transfected with cDNA for human calcitonin receptor showed constitutive activity. Nine ligands for the calcitonin receptor either increased or decreased constitutive activity in this assay. The sensitivity of the system to human calcitonin increased with increasing constitutive activity. These data indicate that, for those receptors which naturally produce constitutive activity, screening in this mode could be advantageous over other methods. PMID- 11033436 TI - Use of recursive partitioning in the sequential screening of G-protein-coupled receptors. AB - High-throughput screening (HTS) is changing as more compounds and better assay techniques become available. HTS is also generating a large amount of data. There is a need to rationalize the HTS process, because, in some cases, the screening of all available compounds is not economically feasible. In addition to the selection of promising compounds, there is a need to learn from the data that we collect. In this paper, we use a data-mining method, recursive partitioning, to help uncover and understand structure-activity relations and to help biology and chemistry experts make better decisions on which compounds to screen next and better characterize. The sequential-screening process is presented and the results of applying that process to 14 G-protein-coupled receptor assays are reported. PMID- 11033437 TI - Pharmacological characterization of receptor-activity-modifying proteins (RAMPs) and the human calcitonin receptor. AB - Receptor-activity-modifying proteins (RAMPs) are a family of single transmembrane domain proteins shown to be important for the transport and ligand specificity of the calcitonin gene-related peptide (CGRP) receptor. In this report, we describe the analysis of pharmacological properties of the human calcitonin receptor (hCTR) coexpressed with different RAMPs with the use of the Xenopus laevis melanophore expression system. We show that coexpression of RAMP3 with human calcitonin receptor changed the relative potency of hCTR to human calcitonin (hCAL) and rat amylin. RAMP1 and RAMP2, in contrast, had little effect on the change of hCTR potency to hCAL or rat amylin. When coexpressed with RAMP3, hCTR reversed the relative potency by a 3.5-fold loss in sensitivity to hCAL and a 19 fold increase in sensitivity to rat amylin. AC66, an inverse agonist, produced apparent simple competitive antagonism of hCAL and rat amylin, as indicated by linear Schild regressions. The potency of AC66 was changed in the blockade of rat amylin but not hCAL responses with RAMP3 coexpression. The mean pK(B) for AC66 to hCAL was 9.4 +/- 0.3 without RAMP3 and 9.45 +/- 0.07 with RAMP3. For the antagonism of AC66 to rat amylin, the pK(B) was 9.25 +/- 0.15 without RAMP3 and 8.2 +/- 0.35 with RAMP3. The finding suggests that RAMP3 might modify the active states of calcitonin receptor in such a way as to create a new receptor phenotype that is "amylin-like." Irrespective of the physiological association of the new receptor species, the finding that a coexpressed membrane protein can completely change agonist and antagonist affinities for a receptor raises implications for screening in recombinant receptor systems. PMID- 11033438 TI - A combination assay for simultaneous assessment of multiple signaling pathways. AB - We have developed an assay in which modulation of two or more signaling pathways can be assessed concurrently by combining reporter gene systems with fluorescent probe technology. The validation of this method was achieved by indirect analysis of adenylyl cyclase activation with the use of a cyclic AMP response element (CRE)-luciferase reporter system in combination with the measurement of calcium mobilization by Calcium Green-1 AM fluorescence on a fluorescent imaging plate reader. To demonstrate the utility of the method in studying the pharmacology of receptors that couple to more than one G protein, Chinese hamster ovary (CHO) cells, which stably expressed both the CRE-luciferase reporter gene and the human pituitary adenylyl cyclase-activating peptide (PACAP) receptor, were treated with PACAP 1-27 and 1-38. Calcium mobilization and the induction of adenylyl cyclase activity in response to each concentration of peptide were assessed in individuals wells. This assay may also be used to screen for ligands of two or more unrelated receptors simultaneously without compromising the assessment of either signaling pathway. To illustrate this point, Rat-1 fibroblasts, which expressed human alpha1A receptors, were cocultured with CRE-luciferase CHO cells, which expressed human GLP-1 receptors. Calcium mobilization elicited by phenylephrine agonism of the alpha1A receptor was assessed in the same assay as GLP-1-induced activation of adenylyl cyclase. The pEC(50) for each agonist was similar to that observed when the cell lines were not cocultured. The number of different receptors that can be screened per well is limited only by the ability to distinguish different reporter gene signals and fluorescent indicators. PMID- 11033439 TI - Development of scintillation-proximity assays for alpha adrenoceptors. AB - Binding assays have long been used to determine compound affinity and selectivity for various seven-transmembrane receptors. Over time, the degree of complexity has significantly reduced, whereas the throughput of the various assays has greatly increased. In this article, we detail the development of a filter-binding assay and a scintillation-proximity assay (SPA) designed to quantify a compound's affinity for the three alpha1-adrenoceptor subtypes, alpha1A, alpha1B, and alpha1D. The various components of the assays such as ease of assay performance, robustness, cost, and generation of radioactive waste are compared and contrasted. On the basis of the results, the SPA offers many advantages of high throughput assay formats over the traditional filter-binding assay. To follow up on the success of the alpha1-adrenoceptor SPA, SPAs for the three alpha2 adrenoceptors were developed and are detailed in this article. Affinity data generated for a select number of alpha2 compounds agree with reported literature values. These assays, like those for alpha1 subtypes, are very amenable to high throughput screening campaigns. In conclusion, scintillation-proximity assays offer significant advantages over filter-binding assays. PMID- 11033440 TI - Discrimination among osteoblasts? Parathyroid hormone analog may reveal site specific differences in mice. PMID- 11033441 TI - Comparison of bone formation responses to parathyroid hormone(1-34), (1-31), and (2-34) in mice. AB - In this study we used a mouse model system to compare the in vivo effects of parathyroid hormone(1-34) [PTH(1-34)] with that of PTH(1-31) or PTH(2-34) analogs. Daily subcutaneous administration of PTH(1-34) for 15 days caused a dose dependent increase in the serum osteocalcin level and bone extract alkaline phosphatase activity, markers of bone formation. PTH(2-34) was much less potent, whereas PTH(1-31) was equipotent in stimulating bone formation parameters in mice. PTH(1-34) caused significant increases in serum calcium (after 4 h) and tartrate-resistant acid phosphatase activity in bone extract (after 4 h), whereas PTH(2-34) and PTH(1-31) were less potent. Because PTH(1-31) caused a smaller increase in bone resorption parameters compared to PTH(1-34), despite similar effects on bone formation parameters, we evaluated the long-term anabolic effects of PTH(1-31) and PTH(1-34) in mice. Weekly evaluations of serum osteocalcin levels demonstrated that daily injections of PTH(1-34) and PTH(1-31) at 80 microg/kg body weight increased serum osteocalcin levels within 1 week of the start of treatment, which were maintained during the entire 22 week treatment. Assessment of bone density at the end of the treatment period with peripheral quantitated computed tomography (pQCT) revealed that PTH(1-34) caused a significantly greater increase in femoral bone density compared to PTH(1-31) at the middiaphysis (18% vs. 9% over vehicle control; p < 0.001). Both PTH(1-34) and PTH(1-31) increased periosteal circumference compared to vehicle (p < 0.01) without a significant difference between the two treatments. In contrast, PTH(1 34) caused a significantly greater reduction in endosteal circumference than PTH(1-31) (p < 0.001). Both analogs significantly increased maximum load and area of moment of inertia over the vehicle group. In conclusion, our findings suggest that PTH(1-34) and PTH(1-31) may exhibit different anabolic effects at the periosteum vs. endosteum in the long bones of mice. PMID- 11033442 TI - Direct stimulation of osteoclastic bone resorption by bone morphogenetic protein (BMP)-2 and expression of BMP receptors in mature osteoclasts. AB - Bone morphogenetic proteins (BMPs) play an important role in various kinds of pattern formation and organogenesis during vertebrate development. In the skeleton, BMPs induce the differentiation of cells of chondrocytic and osteoblastic cell lineage and enhance their function. However, the action of BMPs on osteoclastic bone resorption, a process essential for pathophysiological bone development and regeneration, is still controversial. In this study, we examine the direct effect of BMPs on osteoclastic bone-resorbing activity in a culture of highly purified rabbit mature osteoclasts. BMP-2 caused a dose- and time dependent increase in bone resorption pits excavated by the isolated osteoclasts. BMP-4 also stimulated osteoclastic bone resorption. The increase in osteoclastic bone resorption induced by BMP-2 was abolished by the simultaneous addition of follistatin, a BMP/activin binding protein that negates their biological activity. Just as it increased bone resorption, BMP-2 also elevated the messenger RNA expressions of cathepsin K and carbonic anhydrase II, which are key enzymes for the degradation of organic and inorganic bone matrices, respectively. Type IA and II BMP receptors (BMPRs), and their downstream signal transduction molecules, Smad1 and Smad5, were expressed in isolated osteoclasts as well as in osteoblastic cells, whereas type IB BMPR was undetectable. BMPs directly stimulate mature osteoclast function probably mediated by BMPR-IA and BMPR-II and their downstream molecules expressed in osteoclasts. The results presented here expand our understanding of the multifunctional roles of BMPs in bone development. PMID- 11033443 TI - Structural and cellular changes during bone growth in healthy children. AB - Normal postnatal bone growth is essential for the health of adults as well as children but has never been studied histologically in human subjects. Accordingly, we analyzed iliac bone histomorphometric data from 58 healthy white subjects, aged 1.5-23 years, 33 females and 25 males, of whom 48 had undergone double tetracycline labeling. The results were compared with similar data from 109 healthy white women, aged 20-76 years, including both young adult reference ranges and regressions on age. There was a significant increase with age in core width, with corresponding increases in both cortical width and cancellous width. In cancellous bone there were increases in bone volume and trabecular thickness, but not trabecular number, wall thickness, interstitial thickness, and inferred erosion depth. Mineral apposition rates declined on the periosteal envelope and on all subdivisions of the endosteal envelope. Because of the concomitant increase in wall thickness, active osteoblast lifespan increased substantially. Bone formation rate was almost eight times higher on the outer than on the inner periosteum, and more than four times higher on the inner than on the outer endocortical surface. On the cancellous surface, bone formation rate and activation frequency declined in accordance with a fifth order polynomial that matched previously published biochemical indices of bone turnover. The analysis suggested the following conclusions: (1) Between 2 and 20 years the ilium grows in width by periosteal apposition (3.8 mm) and endocortical resorption (3.2 mm) on the outer cortex, and net periosteal resorption (0.4 mm) and net endocortical formation (1.0 mm) on the inner cortex. (2) Cortical width increases from 0.52 mm at age 2 years to 1.14 mm by age 20 years. To attain adult values there must be further endocortical apposition of 0.25 mm by age 30 years, at a time when cancellous bone mass is declining. (3) Lateral modeling drift of the outer cortex enlarges the marrow cavity; the new trabeculae filling this space arise from unresorbed cortical bone and represent cortical cancelization; (4) Lateral modeling drift of the inner cortex encroaches on the marrow cavity; some trabeculae are incorporated into the expanding cortex by compaction. (5) The net addition of 37 microm of new bone on each side of a trabecular plate results from a <5% difference between wall thickness and erosion depth and between bone formation and bone resorption rates; these small differences on the same surface are characteristic of bone remodeling. (6) Because the amount of bone added by each cycle of remodeling is so small, the rate of bone remodeling during growth must be high to accomplish the necessary trabecular hypertrophy. PMID- 11033444 TI - Quantitative histomorphometric analysis of the human growth plate from birth to adolescence. AB - Longitudinal bone growth occurs via the transformation of growth plate cartilage into bone through a series of cell and matrix changes, termed endochondral ossification. In this study, we characterize the development of trabecular bone from growth plate cartilage in the human rib from birth to adolescence. The height of the proliferative and hypertrophic zones within the growth plate and the primary bone spongiosa decreased with increasing age, with the greatest change observed in the first year of postnatal life. Within these zones, an internal rearrangement of tissue structure occurred. The matrix volume fraction (either cartilage or bone) increased with age in each of the zones. A concomitant increase in cartilage septae thickness and bone trabecular thickness was observed. A decrease in cartilage septae number was seen in the proliferative zone and a decrease in bone trabeculae number was also observed in the primary spongiosa. However, no difference in cartilage septae number was noted in the hypertrophic zone, the region at which cartilage is transformed into bone. Together the proliferative and hypertrophic regions of the growth plate and the bone primary spongiosa appear to constitute the active growth region, with concomitant changes observed that result in longitudinal growth. In contrast, bone mineral volume in the secondary spongiosa was stable over the ages examined; however, trabecular architecture underwent consolidation as trabecular number decreased and trabecular thickness increased. The integration of the structural transformation from cartilage to bone is crucial in achieving the dual purposes of longitudinal growth and peak bone mass. The structure developed during childhood will have an important bearing on the response to bone-altering disease in later life. PMID- 11033445 TI - Expression of P2 receptors in bone and cultured bone cells. AB - Extracellular nucleotides acting through P2 receptors elicit a wide range of responses in many cell types. There is increasing evidence that adenosine triphosphate (ATP) may function as an important local messenger in bone and cartilage. In this study, we used immunocytochemistry, employing novel polyclonal antibodies against P2X(1-7) receptors, and in situ hybridization, using oligonucleotide probes corresponding to P2X(2,4) and P2Y(2,4) messenger RNAs (mRNAs), to localize P2 receptors on undecalcified bone sections and on cultured osteoblasts and osteoclasts. We provide the first direct evidence that the P2X(2) receptor subtype is expressed on osteoclasts, osteoblasts, and chondrocytes. We also obtained evidence for the expression of P2X(5) and P2Y(2) receptors on osteoblasts and chondrocytes, and for P2X(4) and P2X(7) receptors on osteoclasts. Our results confirm earlier reports of P2Y(2) and P2X(4) expression in human osteoclastoma and rabbit osteoclasts, respectively, and are consistent with ATP responses observed on bone cells using electrophysiological techniques. Our novel finding that P2X(2) is expressed by osteoclasts is of particular interest. P2X(2) is the only P2 receptor subtype that requires extracellular acidification to show its full sensitivity to ATP, and our recent functional studies have shown that the stimulatory action of ATP on resorption pit formation by mature osteoclasts is amplified greatly at low pH. These findings point to fundamental new mechanisms for the local modulation of bone resorption. PMID- 11033446 TI - Daily transient decreases in plasma parathyroid hormone levels induced by the calcimimetic NPS R-568 slows the rate of bone loss but does not increase bone mass in ovariectomized rats. AB - Daily parathyroid hormone (PTH) injections that transiently increase plasma PTH levels within the physiological range increase bone mass in osteopenic, ovariectomized (ovx) rats. This study tested the hypothesis that repeated transient decreases in plasma PTH levels from normal, induced by the daily oral administration of the calcimimetic NPS R-568, would induce an anabolic effect in bone of ovx rats with established osteopenia and/or prevent the rapid bone loss that occurs following ovx. In the reversal study, NPS R-568 was administered orally (10 or 100 micromol/kg) for 30 days to 14-month-old retired breeder rats that were ovx 5 months earlier. NPS R-568 treatment did not increase bone formation rate (BFR) or cancellous bone area (B.Ar) in the proximal tibial metaphysis, or bone mineral density (BMD), at any femoral site. In the prevention study, 3-month-old virgin rats were ovx and given NPS R-568 for the following 28 days. The 10 micromol/kg dose prevented the increase in osteoclast number and 42% of the loss of B.Ar, without affecting the elevated osteoblast populations or BFR. Surprisingly, the 100 micromol/kg dose had fewer protective effects, despite preventing the increase in BFR in both cancellous and cortical bone. Detailed analysis of cancellous bone showed that tendency for a dose-related protection of true cancellous bone occurred, but, while the 10 micromol/kg dose prevented 88% of the loss of calcified cartilage seen in control ovx rats, the 100 micromol/kg dose increased that loss by a further 31%. The mechanism underlying these disparate effects of NPS R-568 on calcified cartilage accumulation in the tibial metaphysis is unclear, but may be related to the different effects that the two doses have on plasma Ca(2+) levels. In conclusion, transient increases in PTH levels above basal, and not simple oscillations in hormone levels below normal, appear necessary for the anabolic properties of endogenous PTH to be manifested in the bones of osteopenic ovx rats. PMID- 11033447 TI - Mapping quantitative trait loci for serum insulin-like growth factor-1 levels in mice. AB - Serum insulin-like growth factor-1 (IGF-1) and femoral bone mineral density (BMD) differ between two inbred strains of mice, C3H/HeJ (C3H) and C57BL/6J (B6), by approximately 30% and 50%, respectively. Similarly, skeletal IGF-1 content, bone formation, mineral apposition, and marrow stromal cell numbers are higher in C3H than in B6 mice. Because IGF-1 and several bone parameters cosegregate, we hypothesize that the serum IGF-1 phenotype has a strong heritable component and that genetic determinants for serum IGF-1 are involved in the regulation of bone mass. We intercrossed (B6 x C3H)F1 hybrids and analyzed 682 F2 female offspring at 4 months of age for serum IGF-1 by radioimmunoassay and femoral BMD by peripheral quantitative computerized tomography (pQCT). Genomic DNA was assayed by polymerase chain reaction (PCR) to determine alleles for 114 Mit markers inherited in F2 mice at average distances of 14 centimorgans (cM) along each chromosome (Chr). Serum IGF-1 levels in the F2 progeny were relatively normal in distribution, but showed a greater range than either progenitor, indicating that serum IGF-1 level is a polygenic trait with an estimated heritability of 52%. Serum IGF-1 correlated with femoral length (r = 0.266, p < 0.0001) and femoral BMD (r = 0.267, p < 0.0001). Whole genome scans for main effects associated with serum IGF-1 levels revealed three significant QTLs (in order of significance) on mouse Chrs 6, 15, and 10. The QTL on Chr 6 showed a significant reduction in IGF 1 associated with increasing C3H allele number, whereas the Chr 15 and Chr 10 loci showed additive effects with increasing C3H allele number. A genome-wide search for interacting marker pairs identified a significant interaction between the Chr 6 QTL and a locus on Chr 11. This interactive effect suggested that when the Chr 11 locus was homozygous for C3H, there was no effect of the Chr 6 locus on serum IGF-1; however, the combination of C3H alleles on Chr 6 with B6 alleles on Chr 11 was associated with reduced serum IGF-1 concentrations. To test this in vivo, we tested congenic mice carrying the Chr 6 QTL region from C3H on a B6 background (B6.C3H-6). Both serum IGF-1 and femoral BMD were significantly lower in female congenic than progenitor B6 mice. In summary, we identified three major QTLs on mouse Chrs 6, 10, and 15, and noted a major locus-locus interaction between Chrs 6 and 11. We named these QTLs IGF-1 serum levels (Igf1sl1 to Igf1sl4). Functional isolation of the Igf1sl1 QTL on Chr 6 for IGF-1 in B6.C3H-6 congenic mice demonstrated effects on both the IGF-1 and BMD phenotypes. The genetic determinants of these Igf1sl QTLs will provide much insight into the regulation of IGF-1 and the subsequent acquisition of peak bone mass. PMID- 11033448 TI - Development and evaluation of C-telopeptide enzyme-linked immunoassay for measurement of bone resorption in mouse serum. AB - The mouse is increasingly being used as an animal model for the study of skeletal phenotypes in humans, mainly because of the ease of genetic manipulation. Biochemical markers of bone metabolism provide a valuable parameter for the assessment of skeletal metabolism. In the mouse model, assays for bone formation have been available for a long time; however, little is known about bone resorption markers. The present study describes the development of a serum C telopeptide enzyme-linked immunoassay (ELISA), which measures degradation products of type I collagen that are generated by osteoclastic bone resorption. The C-telopeptide ELISA uses affinity-purified antibodies generated against human sequence DFSFLPQPPQEKAHDGGR. The epitope involves an amino acid sequence, which is identical in the mouse and human C-terminal peptide of type I collagen (alpha1 chain). Sensitivity of the ELISA used was <0.1 ng/mL. The average intra- (n = 10) and interassay (n = 8) coefficient of variation for two controls was <12%. The average dilution and spike recovery rates were 98% and 97%, respectively. Application of the ELISA to measure C-telopeptide in 3-4-week postovariectomized (ovx) C57BL/6J (B6) mice (n = 9 or 10) showed a 45% higher C-telopeptide concentration than the sham-operated mice. Treatment of ovx mice with estradiol (400 microg/kg body weight) or alendronate (1.0 mg/kg body weight) resulted in a 20%-50% decrease in C-telopeptide levels compared to the vehicle-treated ovx group. In addition, B6 mice fed a calcium-deficient diet (0.01% calcium) showed a 50% higher C-telopeptide concentration compared to the B6 mice receiving a normal diet (0.6% calcium). In conclusion, the C-telopeptide ELISA exhibited acceptable analytical performance and sufficient discriminatory power to show expected directional changes in the rate of bone resorption following ovariectomy, ovx plus estradiol or alendronate treatment, and administration of a calcium deficient diet. Therefore, the ELISA developed in this study could be used for measuring bone resorption in the mouse model. PMID- 11033449 TI - Effects of spaceflight and simulated weightlessness on longitudinal bone growth. AB - Indirect measurements have suggested that spaceflight impairs bone elongation in rats. To test this possibility, our laboratory measured, by the fluorochrome labeling technique, bone elongation that occurred during a spaceflight experiment. The longitudinal growth rate (LGR) in the tibia of rats in spaceflight experiments (Physiological Space Experiments 1, 3, and 4 and Physiological-Anatomical Rodent Experiment 3) and in two models of skeletal unloading (hind-limb elevation and unilateral sciatic neurotomy) were calculated. The effects of an 11 day spaceflight on gene expression of cartilage matrix proteins in rat growth plates were also determined by northern analysis and are reported for the first time in this study. Measurements of longitudinal growth indicate that skeletal unloading generally did not affect LGR, regardless of age, strain, gender, duration of unloading, or method of unloading. There was, however, one exception with 34% suppression in LGR detected in slow-growing, ovariectomized rats skeletally unloaded for 8 days by hind-limb elevation. This detection of reduced LGR by hind-limb elevation is consistent with changes in steady-state mRNA levels for type II collagen (-33%) and for aggrecan (-53%) that were detected in rats unloaded by an 11 day spaceflight. The changes detected in gene expression raise concern that spaceflight may result in changes in the composition of extracellular matrix, which could have a negative impact on conversion of growth-plate cartilage into normal cancellous bone by endochondral ossification. PMID- 11033450 TI - A physical, chemical, and mechanical study of lumbar vertebrae from normal, ovariectomized, and nandrolone decanoate-treated cynomolgus monkeys (Macaca fascicularis). AB - Ovariectomized cynomolgus monkeys have previously been investigated as a nonhuman primate model of postmenopausal osteoporosis (Jerome et al., Bone Miner 9:527 540; 1994). In the present study, Fourier transform infrared microspectroscopy (FTIRM) was used to verify that differences in bone mineral quality and quantity in the vertebrae of mature intact (INT) and ovariectomized (ovx) monkeys were analogous to those seen in osteoporotic and nondiseased human bones. FTIRM spectra were acquired from 15 trabeculae per vertebra from three ovx and three INT adult monkeys (mean age 8 years). These spectra were compared with those of both trabecular and previously reported osteonal bone obtained from 3 "normal" and 11 postmenopausal osteoporotic human subjects. While variations in the mineral:matrix ratio (mineral content), carbonate:phosphate ratio, and crystallinity are typical for trabecular bone from iliac crests of normal human subjects, the values of these parameters were relatively static for trabecular bone from postmenopausal osteoporotic human subjects. In general, trabecular bone from postmenopausal osteoporotic human subjects exhibited decreased mineral content (1.0 +/- 0.5 vs. 2.9 +/- 0.6), increased crystallinity, and increased carbonate:phosphate relative to controls. Similarly, trabecular bone from ovariectomized monkeys exhibited lower mineral content (5.8 +/- 0.2) compared with the INT group (6.2 +/- 0.2; p 60 days) have been described in the literature. The pleurisy is associated with pleural and peripheral eosinophilia. Spontaneous regression a few days after withdrawal and radiological cure a few months later is the rule. The precise mechanism of this drug-induced pleural reaction remains unknown. PMID- 11033535 TI - [Wegener's disease mimicking acute infectious pleurisy]. AB - A 75-year-old woman followed for Wegener's disease was hospitalized for pleural effusion. The clinical presentation and cytology of the pleural fluid suggested the diagnosis of purulent pleurisy in this immunodepressed patient. The final diagnosis was a pleural involvement of Wegener's disease, a rarely described localization, as proven by the typical pathology findings. PMID- 11033536 TI - [A mediastinal opacity in a patient with cirrhosis]. AB - Radiological mediastinal shadows have numerous causes. Paraesophageal varices might be revealed by such radiological findings. We described herein the case of a patient with radiological mediastinal opacity occurring after sclerotherapy that was finally related to esophageal varices. PMID- 11033538 TI - [Did you really read this issue? [In Process Citation] PMID- 11033537 TI - [Patient information: guidelines for physicians. ANAES - March 2000. Agence Nationale d'Accreditation et d'Evaluation en Sante]. PMID- 11033539 TI - Founder's Award, Society for Biomaterials. Sixth World Biomaterials Congress 2000, Kamuela, HI,May 15-20, 2000. Really smart bioconjugates of smart polymers and receptor proteins. AB - Over the past 18 years we have been deeply involved with the synthesis and applications of stimuli-responsive polymer systems, especially polymer biomolecule conjugates. This article summarizes our work with one of these conjugate systems, specifically polymer-protein conjugates. We include conjugates prepared by random polymer conjugation to lysine amino groups, and also those prepared by site-specific conjugation of the polymer to specific amino acid sites that are genetically engineered into the known amino acid sequence of the protein. We describe the preparation and properties of thermally sensitive random conjugates to enzymes and several affinity recognition proteins. We have also prepared site-specific conjugates to streptavidin with temperature-sensitive polymers, pH-sensitive polymers, and light-sensitive polymers. The preparation of these conjugates and their many fascinating applications are reviewed in this article. PMID- 11033540 TI - In vitro effects of zirconia and alumina particles on human blood monocyte derived macrophages: X-ray microanalysis and flow cytometric studies. AB - The cytocompatibility of two particulate bioceramics, zirconia and alumina, was studied using human blood monocytes driven to differentiate into mature macrophages with granulocyte macrophage-colony-stimulating factor. Changes in individual cell elemental composition, particularly sodium and potassium content, were assessed by X-ray microanalysis of ultrathin freeze-dried sections. Phagocytosis and respiratory burst of macrophages exposed to biomaterial for 7 days were analyzed under flow cytometry using uptake of fluorescent latex beads and 2'7'-dichlorofluorescien diacetate oxidation, respectively. Zirconia and alumina particles were found to decrease the intracellular potassium/sodium ratio (an index of cell vitality) significantly (p<.01) in 7-day-cultured macrophages compared to control cells cultured out of material. Phagocytosis of both ceramic particles by macrophages was followed by a concomitant decrease in cell phagocytic ability (27%) and a marked altered oxidative metabolism (>2 times reduced by zirconia and >5 times reduced by alumina). The present study clearly demonstrates that reduction of the phagocytic capacity of macrophages associated with altered oxidative metabolism caused by biomaterial particles is characterized by changes in intracellular elemental content. Thus, investigation of cellular homeostasis by electron probe microanalysis together with analysis of functional changes may improve estimation of biomaterial cytocompatibility. PMID- 11033541 TI - The effect of peptide surface density on mineralization of a matrix deposited by osteogenic cells. AB - The density of Arg-Gly-Asp-containing peptides covalently grafted to solid materials has been shown to affect adhesion, spreading, and focal contact formation. The objective of this study was to examine the effect of ligand density on mineralization of the extracellular matrix deposited by osteoblasts. In particular, RGD-modified quartz surfaces with ligand densities varying over two orders (0.01-3.6 pmol/cm(2)) of magnitude were prepared to assess the long term function of osteoblasts on peptide-derivatized surfaces. After 3 weeks in culture, surfaces modified with a 15 amino acid peptide (Ac-Cys-Gly-Gly-Asn-Gly Glu-Pro-Arg-Gly-Asp-Thr-Tyr-Arg-Ala-Tyr-NH(2) ) at a density > or =0.62 pmol/cm(2) significantly (p<0.05) enhanced mineralization compared with a RGD surface density of 0.01 pmol/cm(2), RGE surfaces, or clean surfaces adsorbed with serum proteins. These results suggest that regulation of the surface density of adhesive ligands on biomaterial surfaces is a critical determinant in a strategy to alter the degree of extracellular matrix maturation in contact with solid surfaces (e.g., implants). Further studies are required to elucidate the intracellular signal transduction pathways that mediate long-term matrix mineralization through the initial engagement of these adhesive ligands. PMID- 11033542 TI - Gel-derived materials of a CaO-P(2)O(5)-SiO(2) system modified by boron, sodium, magnesium, aluminum, and fluorine compounds. AB - Bioactive glass-ceramic materials of the CaO-P(2)O(5)-SiO(2) system modified by adding boron, magnesium, sodium, fluorine, and aluminum were obtained using the sol-gel method. Gel-derived materials were produced in the pellet form obtained by compression of powders as well as in coatings on glass slides. The materials obtained were examined in vitro with regard to the ability of calcium phosphate layer to form on the material surface as the result of contact with simulated body fluid (SBF). SBF pH changes and calcium solubility in this solution were determined and scanning electron microscopy, energy-dispersive X-ray analysis, and infrared spectroscopy studies were conducted before and after contact of the materials with SBF. The gels modified by aluminum were amorphous, whereas the sodium and fluorine additives promoted the bulk crystallization of gel-derived materials. The ability of calcium phosphates to crystallize on the surface of gel derived materials depended only slightly on the types of additives applied, and the character of this dependence was different from that observed in melted glasses. Moreover, to estimate the biocompatibility of gel-derived coatings, we examined the proliferation, collagen synthesis, adhesion, and morphology of fibroblasts (NRK cells) cultured in the presence of gel-derived materials. The results of these experiments showed that none of the tested materials significantly reduced any cell function. PMID- 11033543 TI - Messenger ribonucleic acid expression of 16 matrix metalloproteinases in bone implant interface tissues of loose artificial hip joints. AB - Matrix metalloproteinases (MMPs) have been reported to be the major factors responsible for aseptic loosening of artificial hip joints. So far, messenger ribonucleic acid (mRNA) expression patterns of seven MMPs have been reported, but that of many other MMPs which have been newly discovered or recently considered to be responsible for prosthetic loosening is still unknown. In this study, mRNA expression pattern of 16 different types of MMPs were analyzed to evaluate which MMPs were locally produced and contributed to prosthetic loosening. Synovium-like interface tissues between bone and prosthesis were collected from 18 cases of aseptic loose artificial hip joint at revision surgery. Six cases of normal synovium were used as controls. Total RNA was extracted by single-step acid guanidinium-thiocyanate-phenol-chloroform procedure. mRNA expression of MMPs was analyzed by semiquantitative reverse transcription-polymerase chain reaction. Based on local expression pattern of MMPs at the mRNA level, aseptic loose artificial hip joint was characterized by elevated expression of MMP-1, MMP-9, MMP-10, MMP-12, and MMP-13; moderate expression of MMP-2, MMP-7, MMP-8, MMP-11, membrane type (MT)1-MMP (MMP-14), MT2-MMP (MMP-15), MT3-MMP (MMP-16), MT4-MMP (MMP-17), and MMP-19; lower expression of MMP-3; and little significance of MMP 20. The MMPs detected in this study can potentially degrade almost all components of the periprosthetic extracellular matrix. Thus, many MMP type enzymes possibly contribute to prosthetic loosening and osteolysis through pathologic extracellular matrix degradation and connective tissue/bone remodeling around prostheses. PMID- 11033544 TI - Enhancement of the in vivo osteogenic potential of marrow/hydroxyapatite composites by bovine bone morphogenetic protein. AB - A composite of marrow mesenchymal stem cells and porous hydroxyapatite (HA) has in vivo osteogenic potential. To investigate factors enhancing the osteogenic potential of marrow/HA composites, we prepared a bone morphogenetic protein (BMP) fraction from the 4M guanidine extract of bovine bone by heparin-sepharose affinity chromatography. Marrow/HA composites or composites containing marrow mesenchymal stem cells, BMP, and HA (marrow/BMP/HA composites) were implanted subcutaneously in 7-week-old male Fischer rats. BMP/HA composites and HA alone were also implanted. The implants were harvested after 2, 4, or 8 weeks and were prepared for histological and biochemical studies. Histological examination showed obvious de novo bone formation together with active osteoblasts at 2 weeks, as well as more extensive bone formation at 4 and 8 weeks in many pores of the marrow/BMP/HA composites. The marrow/HA composites did not induce bone formation at 2 weeks, but there was moderate bone formation at 4 weeks. At 2 weeks, only marrow/BMP/HA composites resulted in intensive osteogenic activity, judging from alkaline phosphatase and osteocalcin expression at both the protein and gene levels. These results indicate that the combination of marrow mesenchymal stem cells, porous HA, and BMP synergistically enhances osteogenic potential, and may provide a rational basis for their clinical application, although further in vivo experiment is needed. PMID- 11033545 TI - Fibrous tissue ingrowth and attachment to porous tantalum. AB - This study determined the soft tissue attachment strength and extent of ingrowth to a porous tantalum biomaterial. Eight dorsal subcutaneous implants (in two dogs) were evaluated at 4, 8, and 16 weeks. Upon retrieval, all implants were surrounded completely by adherent soft tissue. Implants were harvested with a tissue flap on the cutaneous aspect and peel tested in a servo-hydraulic tensile test machine at a rate of 5 mm/min. Following testing, implants were dehydrated in a solution of basic fuschin, defatted, embedded in methylmethacrylate, and processed for thin-section histology. At 4, 8, and 16 weeks, the attachment strength to porous tantalum was 61, 71, and 89 g/mm respectively. Histologic analysis showed complete tissue ingrowth throughout the porous tantalum implant. Blood vessels were visible at the interface of and within the porous tantalum material. Tissue maturity and vascularity increased with time. The tissue attachment strength to porous tantalum was three- to six-fold greater than was reported in a similar study with porous beads. This study demonstrated that porous tantalum permits rapid ingrowth of vascularized soft tissue, and attains soft tissue attachment strengths greater than with porous beads. PMID- 11033546 TI - Peripheral nerve regeneration using bioresorbable macroporous polylactide scaffolds. AB - The ability of DRG-derived neurons to survive and attach onto macroporous polylactide (PLA) foams was assessed in vitro. The foams were fabricated using a thermally induced polymer-solvent phase separation. Two types of pore structures, namely oriented or interconnected pores, can be produced, depending on the mechanism of phase separation, which in turn can be predicted by the thermodynamics of the polymer-solvent pair. Coating of the porous foams with polyvinylalcohol (PVA) considerably improved the wettability of the foams and allowed for cell culture. The in vitro biocompatibility of the PVA-coated supports was demonstrated by measuring cell viability and neuritogenesis. Microscopic observations of the cells seeded onto the polymer foams showed that the interconnected pore networks were more favorable to cell attachment than the anisotropic ones. The capacity of highly oriented foams to support in vivo peripheral nerve regeneration was studied in rats. A sciatic nerve gap of 5-mm length was bridged with a polymer implant showing macrotubes of 100 microm diameter. At 4 weeks postoperatively, the polymer implant was still present. It was well integrated and had restored an anatomic continuity. An abundant cell migration was observed at the outer surface of the polymer implant, but not within the macrotubes. This dense cellular microenvironment was found to be favorable for axogenesis. PMID- 11033548 TI - A mechanistic study of the antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus. AB - To investigate the mechanism of inhibition of silver ions on microorganisms, two strains of bacteria, namely Gram-negative Escherichia coli (E. coli) and Gram positive Staphylococcus aureus (S. aureus), were treated with AgNO(3) and studied using combined electron microscopy and X-ray microanalysis. Similar morphological changes occurred in both E. coli and S. aureus cells after Ag(+) treatment. The cytoplasm membrane detached from the cell wall. A remarkable electron-light region appeared in the center of the cells, which contained condensed deoxyribonucleic acid (DNA) molecules. There are many small electron-dense granules either surrounding the cell wall or depositing inside the cells. The existence of elements of silver and sulfur in the electron-dense granules and cytoplasm detected by X-ray microanalysis suggested the antibacterial mechanism of silver: DNA lost its replication ability and the protein became inactivated after Ag(+) treatment. The slighter morphological changes of S. aureus compared with E. coli recommended a defense system of S. aureus against the inhibitory effects of Ag(+) ions. PMID- 11033547 TI - The effect of alkali- and heat-treated titanium and apatite-formed titanium on osteoblastic differentiation of bone marrow cells. AB - This study was based on the hypothesis that osteogenesis is enhanced by growth of osteogenic cells on an apatitic surface. To test this hypothesis, the behavior of rat bone marrow cells on these surfaces was examined: commercially pure titanium (Cp Ti), alkali- and heat-treated titanium (AH Ti), and AH Ti incubated in a simulated body fluid to deposit crystalline hydroxyapatite on the surface (Ap Ti). The alkaline phosphatase (ALP) activity of the cells cultured on Ap Ti was significantly higher at day 7 and day 14 than the ALP activity observed for the other titanium surfaces. At day 14, the ALP activity on AH Ti was significantly increased compared with the ALP activity on Cp Ti. The amount of DNA per well increased nearly in parallel for each titanium. However, northern blot analysis at day 14 revealed that expression of osteocalcin and alpha1(I) collagen mRNA was higher in the cells cultured on Ap Ti than the cells cultured on AH Ti. The cells cultured on Cp Ti showed the lowest mRNA levels. After 7 days of cell-free culture in medium supplemented with 15% serum, X-ray photoelectron spectroscopy (XPS), and thin-film X-ray diffraction (TF-XRD) analysis showed that calcium phosphate had been deposited on the AH Ti (resulting in an increase in thickness with time). No phosphate was detected on the Cp Ti, even after day 14. This study indicates that Ap Ti provides the most favorable conditions for differentiation of bone marrow cells, and, at a later stage, AH Ti also provides favorable conditions, perhaps because of the formation of a surface layer of calcium phosphate. This potential for apatite formation may play an important role in osteoblastic differentiation. PMID- 11033549 TI - Type I atelocollagen grafting onto ozone-treated polyurethane films: cell attachment, proliferation, and collagen synthesis. AB - An approach is presented for the graft copolymerization of type I atelocollagen onto the surface of polyurethane (PU) films treated with ozone. Through inducing oxidization to modify PU surface by ozone, peroxide groups are easily generated on the surface. Those peroxides are broken by redox-polymerization, and provide active species which initiate graft polymerization by reacting with amines in the collagen molecules. The ozone oxidation time and voltage could readily control the amount of peroxide production. The surface density of generated peroxides on PU surface was determined by iodide method. The maximum concentration of peroxide was about 10.20 x 10(-8)mol/cm(2) when ozone oxidation was performed at 60 V for 30 min. After the reaction of PU by ozone oxidation, type I atelocollagen was graft-copolymerized onto the PU film. All the physical measurements on the collagen-grafted surface indicated that the PU surface was effectively covered with type I atelocollagen. The interaction of the collagen-grafted PU surface with fibroblasts could be greatly enhanced by the surface graft polymerization with type I atelocollagen. Attachment and proliferation of fibroblasts on the grafted type I atelocollagen were significantly enhanced, and it is assumed that the atelocollagen matrix supported the initial attachment and growth of cells. In the early stage of proliferation, collagen synthesis in fibroblasts was not activated and remained at a relatively low level due to the grafted type I atelocollagen, increasing only with fibroblast differentiation. PMID- 11033550 TI - Mechanical, thermal, and chemical analyses of the binary system Au-Ti in the development of a dental alloy. AB - Investigations in the binary system gold-titanium (Au-Ti) were performed in regard to the development of a universal dental alloy suited for inlays, for the conventional crown and bridge technique, as well as for the ceramic-fused-to metal technique. With nine alloys with increasing Ti content from 0-10 atom %, microstructure, mechanical properties, thermal properties, and corrosion were determined in the as-cast state and after simulation of ceramic firing. The microstructure shows an increasing formation of a second phase with increasing Ti content, crystallizing dendritically, which, according to the binary phase diagram must be the intermetallic phase TiAu(4). The results of the measurements reveal that with increasing Ti content, hardness, Young's modulus, proof stress, and tensile strength strongly increase in the range of 2-6 atom % Ti and with higher Ti content remain constant. Elongation after fracture decreases with a Ti content above 2 atom %. The coefficient of thermal expansion decreases linearly with increasing Ti content. Thermal distortion decreases exponentially with increasing Ti content. The corrosion test showed an increasing release of Ti with increasing Ti content. Lowest corrosion was observed in the simulated state with intact oxide layer. Appropriate properties for dental application were found in the range of 6.5 at% Ti. PMID- 11033551 TI - Characterization of bioactive glass-reinforced HAP-polymer composites. AB - The effect of bioactive glass on the mechanical properties of hydroxyapatite-Ca polyacrylate composites was studied. Powder mixtures of tetracalcium phosphate (TetCP), poly(acrylic-co-itaconic) and bioactive glass (up to 50% by weight) were hot pressed for 30 min at 300 degrees C and 40 kpsi. Tensile strengths, elastic moduli, and microstructures of the composites produced were investigated. Results showed the mechanical properties of these composites were enhanced by the addition of bioactive glass. The highest values of tensile strength and elastic modulus were achieved with the addition of 10% bioactive glass. Composites were immersed in SBF for up to 10 days, then in 1.5 simulated body fluid (SBF) for a week. The changes in the concentrations of Ca, P, and Si ions of these solutions were measured. The microstructures of these composites after SBF immersion were also evaluated. Concentrations of Ca, P, and Si increased with the time of immersion in SBF owing to the formation of an apatite layer on their surfaces as found by SEM with energy-dispersive spectroscopy attachment. PMID- 11033552 TI - Chemico-physical properties of hyaluronan-based sponges. AB - The aim of this study was to obtain information on the chemico-physical and surface properties of the hyaluronan total benzylic ester sponges to evaluate their stability, surface "cleanliness" and handling for the applications in the tissue engineering. The thermal analysis, the characterization of surface chemical composition and the swelling test were performed on these materials. Moreover, the morphological changes, the rheological behavior, and the molecular weight loss in function of the time were monitored when the sponges were incubated in cell culture medium. The results showed that the sponges were thermally stable up to 220 degrees C and the surface composition was different from that of the bulk for C-O contribution. No contaminants were detected. In culture medium, the samples swelled assuming the rheological properties of biopolymer gel. When the sponges were in contact with the culture medium, their molecular weight remained stable for the first day and a loss of 11% and 31% was recorded for samples removed from culture medium after 3 and 7 days, respectively. With the scanning electron microscopy analysis, the spongy structure appeared with open interconnecting pores. The micrographs related to the samples after incubation in culture medium showed that the degradation was evident on the surface after 1 day. The deterioration of the pore walls and the presence of craters increased with time and, after 3 days, the phenomena were present also in the section. PMID- 11033553 TI - Semi-interpenetrating polymer networks composed of biocompatible phospholipid polymer and segmented polyurethane. AB - 2-Methacryloyloxyethyl phosphorylcholine (MPC) polymers, which have excellent biocompatibility, have been receiving increasing attention in biomedical and bioengineering fields; however, the mechanical strength of the hydrated MPC polymers is not sufficient for use in these fields as a bulk material. Therefore, we hypothesized that a novel material might be realized by reinforcing the MPC polymer network with segmented polyurethane (SPU). Semi-interpenetrating polymer networks (IPNs) composed of crosslinked MPC polymer and SPU were prepared. The mechanical properties of the IPN membrane were significantly improved compared with those of the MPC polymer membrane. Three-dimensional polymer networks of the MPC polymer in the IPNs were observed after solvent extraction of SPU. An X-ray photoelectron spectrum analysis revealed that the MPC units were exposed on the IPN surface. When the IPN was alternately soaked in water and ethanol, the swelling ratio was found to be completely reversible and no disintegration of the network structure was observed. The permeation coefficient of 1, 4-di(2 hydroxyethoxy)benzene through the IPN membrane was 1.11 x 10(-7) cm(-2)s(-1). The amount of adsorbed protein and the number of adherent platelets on the IPN membrane were effectively reduced compared with those on SPU. We concluded that IPNs composed of the MPC polymer and SPU are a new bulk biomaterial, which possesses both blood compatibility and good mechanical properties. PMID- 11033554 TI - Prevention of infection with tobramycin-containing bone cement or systemic cefazolin in an animal model. AB - We investigated in an animal model the efficacy of tobramycin-containing bone cement and systemic cefazolin for infection prophylaxis. In 18 female rabbits, the femoral cavity was inoculated with Staphylococcus aureus before injection of bone cement. The first group of six rabbits received tobramycin-containing Simplex-P bone cement. Two other groups of six rabbits received plain Simplex-P bone cement. Preoperatively, in one of the two latter groups cefazolin was administered intravenously. The other group served as untreated controls. The rabbits were monitored for clinical signs of infection. At 7 days' follow-up, the femora were harvested and cultures from the bone adjacent to the cement plug were quantified. Cultures from the rabbits which received antibiotic prophylaxis (either cefazolin systemically or tobramycin-containing bone cement) were all negative. In contrast, all rabbits in the untreated control group had positive cultures. These rabbits also had other signs of infection such as an elevated erythrocyte sedimentation rate and loss of body weight. Culture results were confirmed by the absence of bacterial DNA in the polymerase chain reaction hybridization assay. In conclusion, we found that both tobramycin-containing bone cement and systemic cefazolin are effective in preventing implant bed infection in rabbits up to 7 days after contamination with S. aureus. PMID- 11033555 TI - Beta-1 integrin expression by human nasal chondrocytes in microcarrier spinner culture. AB - Beta-1 integrin plays a major role in cell attachment and is believed to be involved in mediating the interactions of chondrocytes with their environment. We previously reported that articular chondrocytes propagated in microcarrier spinner culture proliferated and reexpressed their chondrocytic protein. The goal of the present study was to investigate the expression of beta-1 integrin by chondrocytes growing on the surface of microcarriers. Nasal chondrocytes (4 x 10(3)/cm(2)) were seeded on microcarriers and incubated at 37 degrees C, 5% CO(2), 60 rpm. Expression of chondrocyte markers and beta-1 integrin was determined using reverse transcriptase-polymerase chain reaction and immunocytochemical analyses. De novo synthesis of sulfate-containing proteoglycans was studied using 35SO(4) incorporation techniques. Like articular chondrocytes propagated in microcarrier spinner culture, nasal chondrocytes expressed high levels of collagen type II mRNA, whereas collagen type I mRNA levels were low. Aggrecan mRNA was detectable and levels of de novo 35SO(4) incorporation were high. Chondrocytes immunostained intensely for collagen type II and keratan sulfate but did not stain for collagen type I. beta-1 integrin mRNA levels were high, and the protein was immunolocalized to regions of cell-to cell or cell-to-microcarrier contact. The fact the chondrocytes expressed high levels of beta-1 integrin raises the possibility that this integrin molecule has a role in the maintenance of the chondrocytic phenotype. PMID- 11033556 TI - Integrin-mediated signaling regulates AP-1 transcription factors and proliferation in osteoblasts. AB - Since osteoblast proliferation is critical for bone development, the effect of bone extracellular matrix (ECM) proteins on osteoblast signaling and proliferation in serum-free medium was investigated. Proliferation was highest in primary rat calvarial osteoblasts cells grown on fibronectin but less on type I collagen; osteonectin and poly-L-lysine did not support early proliferation. Fibronectin and type I collagen binding requires integrins, whereas cell adhesion to osteonectin or poly-L-lysine does not involve integrins. Therefore, the role of integrins in osteoblast signaling, leading to the induction of AP-1 transcription factors (c-fos and c-jun) which are important in cell proliferation, was studied. c-fos and c-jun message levels were increased at 60 min in osteoblasts plated onto fibronectin or collagen, but not in cells on osteonectin or poly-L-lysine. Protein synthesis was not required for c-fos mRNA expression; however, kinase activity was necessary for c-fos induction. In cells plated onto fibronectin, c-fos mRNA levels were controlled by protein kinase C and phosphotyrosine kinase signaling pathways. In contrast, c-fos levels in collagen-adhering cells may involve protein kinase A. The signaling pathway involving the phosphorylation of focal adhesion kinase and mitogen-activated kinases was also shown to be transiently increased in osteoblasts on fibronectin and type I collagen, but not in cells on poly-L-lysine. These results demonstrate that osteoblast binding to the extracellular matrix through integrins induces c fos and c-jun, and that both fibronectin and collagen affect these AP-1 transcription factors through protein kinase-sensitive pathways. Thus, osteoblast proliferation is modulated differentially by specific ECM components. PMID- 11033557 TI - Integrin-mediated signaling in osteoblasts on titanium implant materials. AB - The intracellular signaling pathway for osteoblast adhesion to the orthopedic implant material Ti6Al4V (TIV) was investigated and compared to integrin-mediated adhesion to extracellular matrix proteins. Primary osteoblasts from fetal rat calvaria were plated onto TIV, fibronectin (FN), and poly-L-lysine (PLL) and the levels of focal adhesion kinase (FAK), mitogen-activated protein kinase (MAPK), and AP-1 transcription factors, c-fos and c-jun, were compared by Western and Northern blots. Cells on all substrates showed maximum FAK phosphorylation within 60 min and then a decrease at 2 and 24 h. However, the subsequent signal transduction pathway differed on PLL compared to TIV and FN. MAPK was phosphorylated similarly in osteoblasts attached to FN and TIV, whereas cells on PLL demonstrated no MAPK phosphorylation. On TIV and FN, c-fos and c-jun mRNA levels were maximal within 1 h and then plateaued or declined by 2 h. On PLL, they increased at 2 h. Within 1 h, c-fos protein was stimulated in cells attached to TIV and FN and decreased in cells on PLL. c-jun protein increased on all substrates compared to unplated cells. Cytoskeletal changes visualized by phalloidin fluorescence microscopy at 4 h of culture were delayed on TIV compared to FN. In addition, approximately 50% fewer cells adhered to TIV compared to FN or PLL. By 24 h, a well-spread cytoskeleton with focal adhesion sites was apparent on TIV and FN, but cells on PLL were rounded with minimal cell spreading. During 6 days of culture, cells on FN and TIV proliferated, whereas the number of cells on PLL remained the same or decreased, depending on the initial plating density. We conclude that osteoblast adhesion to TIV implants is similar to osteoblast adhesion to FN and leads to osteoblast proliferation. These data provide evidence for the biocompatibility of TIV at a molecular level. PMID- 11033558 TI - The role of cell density in the survival of cultured cerebellar granule neurons. AB - The dependence for survival of cerebellar granule neurons on the cell density was examined both experimentally and theoretically. The results of batch experiments revealed that the cell survival index (CSI) was inappreciable, if cell density was below a critical level. If cell density exceeded this critical value, CSI increased with the increase in cell density. In addition, CSI was significantly increased by using a conditioned medium from the dense cultures. This suggests that not only cell density promotes survival of neurons, but also an increased concentration of growth factors produced by neurons has a direct effect on the survival of the neurons. A quantitative model describing the distribution of the growth factor at different cell densities was proposed to investigate the role of cell density in the survival of the neurons. We showed the existence of a critical level for cell density, and good agreement in the improvement of CSI was found between the theoretical prediction and the experimental result. Finally, the average concentration of growth factor necessary for cell survival based on our model was in a reasonable range compared to the practice of the addition of neurotrophic factors to the medium of cultured cerebellar granule neurons. PMID- 11033559 TI - Prophylaxis of implant-related staphylococcal infections using tobramycin containing bone cement. AB - In a rabbit model, premixed tobramycin-containing bone cement was studied for its efficacy to prevent infections with two frequently encountered staphylococcal species in arthroplasty surgery. After intramedullary inoculation with staphylococci, either standard or premixed tobramycin-containing Simplex-P bone cement was injected in the right femur of 120 rabbits. Development of infection was examined by culture of femoral bone after 7 or 28 days. Loss of body weight and elevated erythrocyte sedimentation rate in the control rabbits inoculated with Staphylococcus aureus were seen in the first postoperative week, returning to normal in 28 days. Inoculation with Staphylococcus epidermidis resulted only in a low-grade infection. All rabbits receiving premixed tobramycin-containing bone cement were free of signs of infection, and all their cultures were negative. Culture yield from Staphylococcus aureus controls increased with time and inoculum dose. Staphylococcus epidermidis controls needed higher inoculum doses to establish an infection, while culture yield decreased in time. These differences in mode of prosthesis-related infection are explained by differences in virulence factors. PMID- 11033561 TI - Plasma-based sterilization: effect on surface and bulk properties and hydrolytic stability of reprocessed polyurethane electrophysiology catheters. AB - Plasma-based sterilization is a promising alternative to ethylene oxide (EO) for reprocessing of electrophysiology catheters. To assess its safety in terms of material damage, modifications of surface and bulk properties as well as hydrolytic stability of sterilized catheters were evaluated. Polyurethane (PU) single-use electrophysiology catheters were subjected to one, five, and ten sterilization cycles by Sterrad-100S and Plazlyte, as well as by pure EO for comparison. Surface analysis techniques (ATR-FTIR, XPS, DCA) showed oxidation limited to the near-surface layer induced by both plasma-based sterilizers, whereas EO induced slight but deeper alkylation. Using bulk analysis techniques (RP-HPLC, SEC), oligomer alteration was observed after all three sterilization techniques, without modification of molecular weights. Hydrolytic stability of catheters was slightly changed by plasma-based sterilization, with a small increase in released oligomers. Finally, although Plazlyte and Sterrad are both plasma-based techniques, they induced different impacts on catheters, such as the degradation of an additive with Sterrad, and a clear difference in coloration with Plazlyte. PMID- 11033560 TI - Effects of amelogenin on the transforming surface microstructures of Bioglass in a calcifying solution. AB - Topographies of a bioactive glass (45S5 type Bioglass(R)) during 0-4 h of immersion in a supersaturated calcifying solution (SCS) and the SCS containing recombinant porcine amelogenin rP172 (SCS(rP172)) were observed by atomic force microscopy. Other techniques including X-ray diffraction, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, and transmission electron microscopy were used for some complementary microstructural investigations. The smooth Bioglass surface changed to be very rough after 0.5 h of SCS immersion because of glass network dissolution. Spherical silica-gel particles with diameters of 150-300 nm consisting of substructures of 20-60 nm across had formed on the sample surfaces after 1 h of SCS immersion. The chemisorption of amorphous calcium phosphate and crystallization of nanophase apatite were seen to occur epitaxially on the silica-gel structures during 1-4 h of SCS immersion. During the first 0.5 h of SCS(rP172) immersion, more than 95% of rP172 protein in solution was adsorbed onto the sample surfaces and generated spherical assemblies of 10-60 nm diameters. During 0.5-4 h of SCS(rP172) immersion, the protein assemblies of rP172 remarkably induced the formation of orientated silica-gel plates (approximately 100-nm wide and 50-nm thick) and subsequently of long and thin apatite needle crystals. The recombinant amelogenin rP172-modulated apatite crystals resembled those formed in the early stage of tooth enamel biomineralization, suggesting the functional roles of amelogenins during the oriented growth of enamel crystallites and a great potential for amelogenins in applications designed to fabricate enamel-like calcium phosphate biomaterials. PMID- 11033562 TI - Effect of varying physical properties of porous, surface modified bioactive glass 45S5 on osteoblast proliferation and maturation. AB - The objective of this study was to determine the effect of porous bioactive glass (45S5) substrate characteristics on the expression and maintenance of the osteoblastic phenotype. We cultured ROS 17/2. 8 cells on substrates with different pore size and porosity for periods up to 14 days and analyzed the characteristics of the cells and extracellular matrix. Results of the study show that the glass substrates supported the proliferation and growth of osteoblast like cells. Although the morphologies of the cells differed on the various substrates, their shape and the extent of membrane ruffling suggested that they maintained high levels of metabolic activity. Cells on all substrates expressed high levels of alkaline phosphatase activity and produced extracellular matrices that mineralized to form nonstoichiometric, carbonated, calcium-deficient apatites. An important finding was that at a given porosity of 44%, the pore size neither directed nor modulated the in vitro expression of the osteoblastic phenotype. In contrast, porosity did affect cellular function. We noted that at an average pore size of 92 microm, as the porosity increased from 35 to 59%, osteoblast activity was reduced. As designed in this experiment, an increase in the porosity led to a corresponding increase in total surface area of the specimens. With increasing porosity and surface area, glass reactions in the media may persist for longer durations at higher intensities, thereby affecting local media composition. As such, we suggest that extensive conditioning treatments before cell seeding can reduce this effect. Our results also revealed that the expression of the osteoblastic phenotype is enhanced by the ongoing glass dissolution. The reaction pathway at the origin of this effect still needs to be elucidated. Taken together, the findings support the overall hypothesis that in vitro cell activity can be controlled by a careful selection of substrate properties. PMID- 11033563 TI - Resistance to sliding of orthodontic appliances in the dry and wet states: influence of archwire alloy, interbracket distance, and bracket engagement. AB - Having established dimensional and mechanical characteristics, the resistances to sliding (RS) were measured in vitro for various archwires against stainless steel brackets. Using stainless steel ligatures, a constant normal force (300g) was maintained while second-order angulation (straight theta) was varied from -12 degrees to +12 degrees. Using miniature bearings to simulate contiguous teeth, five experiments each were run in the dry or wet states with human saliva at 34 degrees C as a function of four archwire alloys, five interbracket distances, and two bracket engagements. Outcomes were objectively analyzed to establish when theta=0, and the relative contact angles ( theta(r)) were replotted. Critical contact angles (theta(c)) that were determined via experimentation were in good agreement with theory. Slopes and y-intercepts were tabulated from linear regression equations of RS against theta plots in both the passive (theta < or = theta (c)) and active ( theta > or = theta(c)) configurations, for which theta = theta(c) identified the boundary between classical friction and binding phenomena. Stiffer archwires and shorter interbracket distances exacerbated binding, whereas, once corrected for differing bracket engagement, RS was independent of slot dimension. Unlike earlier results in the passive configuration, in the active configuration couples comprised of titanium alloys (NiTi and (beta-Ti) had higher RS values in the wet versus the dry state. For those archwire alloys evaluated, two empirical expressions were adduced that comprise the binding component, the yield strength or elastic limit, and the beam length, which implicitly represent the stiffness, flexibility, and interbracket distance. PMID- 11033564 TI - Whisker-reinforced dental core buildup composites: effect of filler level on mechanical properties. AB - The strength and toughness of dental core buildup composites in large stress bearing restorations need to be improved to reduce the incidence of fracture due to stresses from chewing and clenching. The aims of the present study were to develop novel core buildup composites reinforced with ceramic whiskers, to examine the effect of filler level, and to investigate the reinforcement mechanisms. Silica particles were fused onto the whiskers to facilitate silanization and to roughen the whisker surface for improved retention in the matrix. Filler level was varied from 0 to 70%. Flexural strength, compressive strength, and fracture toughness of the composites were measured. A nano indentation system was used to measure elastic modulus and hardness. Scanning electron microscopy (SEM) was used to examine the fracture surfaces of specimens. Whisker filler level had significant effects on composite properties. The flexural strength in MPa (mean +/- SD; n = 6) increased from (95+/-15) for the unfilled resin to (193+/- 8) for the composite with 50% filler level, then slightly decreased to (176+/-12) at 70% filler level. The compressive strength increased from (149+/-33) for the unfilled resin to (282+/-48) at 10% filler level, and remained equivalent from 10 to 70% filler level. Both the modulus and hardness increased monotonically with filler level. In conclusion, silica particle-fused ceramic single-crystalline whiskers significantly reinforced dental core buildup composites. The reinforcement mechanisms appeared to be crack deflection and bridging by the whiskers. Whisker filler level had significant effects on the flexural strength, compressive strength, elastic modulus, and hardness of composites. PMID- 11033565 TI - Calcium level-responsive in-vitro zinc release from zinc containing tricalcium phosphate (ZnTCP). AB - The in vitro Zn release from tricalcium phosphate containing Zn (ZnTCP; 0.63, 6.17, and 12.05 Zn w/w%) was investigated. The rates of release from ZnTCP powders were measured in 25 mL of simulated body fluid (SBF) containing 10 mg/100mL Ca (SBF/H), 5 mg/100mL Ca (SBF/L), or no Ca (SBF/-) at pH 7.25, 37.0+/ 0.1 degrees C. The release from 6 and 12% ZnTCP was initially very fast. The rate of release from ZnTCP decreased as the concentration of Ca in the dissolution media increased, but increased as concentration of Zn in TCP increased. The dissolution kinetics of ZnTCP followed the Hixon-Crowell equation at the initial stage of dissolution, and the initial dissolution rate constant (IDR) was calculated by the least-squares method. The effect of Ca concentration on percent IDR of ZnTCP suggested that Zn release from 0.6% ZnTCP was significantly high compared to that from 6 or 12% ZnTCP. The relationship between the amount of Ca precipitated and Zn release of various ZnTCP samples suggested that the release from 0.6% ZnTCP was significantly different compared to that from 6 and 12% ZnTCP, consistent with the data for percent Zn IDR. X-ray diffraction data suggested that 0.6% ZnTCP contained 5% hydroxyapatite, a low solubility material, which acted as seed crystal during the dissolution test. PMID- 11033566 TI - Initial events at the bioactive glass surface in contact with protein-containing solutions. AB - Upon implantation, bioactive glass undergoes a series of reactions that leads to the formation of a calcium phosphate-rich layer. Most in vitro studies of the changes that occur on the surface of bioactive glass have employed the use of buffer solutions with compositions reflecting the ionic composition of interstitial fluid. Although these studies have documented the physical and chemical changes associated with bioactive glass immersed in aqueous media, they do not reveal the effect of serum proteins and cells that are present at the implantation site. In the present study, we document, using atomic force microscopy (AFM) and Rutherford backscattering spectrometry (RBS), significant differences in the reaction layer composition, thickness, morphology, and kinetics of formation arising from the presence of serum proteins. The data reveal that the uniform and rapid adsorption of serum proteins on the surface may serve to protect the surface from further direct interaction with the aqueous media, slowing down the transformation reactions. This finding is in agreement with previous studies that have shown that the presence of serum proteins significantly delays the formation of hydroxyapatite at the surface of bioactive glass. These data also support the hypothesis that initial reaction layers in vivo interact with cells in order to produce the tissue-bioactive glass interface typically observed on ex vivo specimens. PMID- 11033567 TI - Micelles of poly(ethylene oxide)-block-poly(N-alkyl stearate L-aspartamide): synthetic analogues of lipoproteins for drug delivery. AB - Stearic acid esters of poly(ethylene oxide)-block-poly(hydroxyethyl L aspartamide) and poly(ethylene oxide)-block-poly(hydroxyhexyl L-aspartamide) have been synthesized from poly(ethylene oxide)-block-poly(beta-benzyl L-aspartate) by polymer-analogous reactions and self-assembled into a micelle. Transmission electron microscopy and fluorescent probe studies reveal that the micelle mimics structural features of serum lipoproteins: it is nanoscopic, spherical, and has a supramolecular core-shell architecture, where the core is rich in fatty acid esters. As a result, the polymeric micelles effectively solubilize amphotericin B, a key drug for systemic mycoses. Serum lipoproteins solubilize many hydrophobic drugs as a biological transport system besides amphotericin B. A synthetic polymeric analogue may achieve the same aim, but with the ease of structural modification, safety, and stability. PMID- 11033568 TI - Preparation of a PEG-grafted phospholipid Langmuir-Blodgett monolayer for blood compatible material. AB - In order to develop a versatile model for blood-compatible materials, we studied morphology and platelet adhesion of a dipalmitoyl-phosphatidylcholine (DPPC)/dipalmitoyl-phosphatidylethanolamine-polyethylene glycol (PEG lipid) mixed monolayer. This monolayer, which mimics the cell membrane structure, consists of two heterogeneous layers, that is, a PEG layer lying on top of a phospholipid monolayer. The DPPC/PEG lipid mixed monolayer was prepared using the Langmuir Blodgett (LB) Technique. The monolayer was transferred onto a silanized glass substrate by the down-stroke mode, at a surface pressure of 25 mN/m. The transfer efficiency achieved unity at all times. The morphologies of PEG chains on the phospholipid monolayer in water, in a dried state, and in a hydrated state were evaluated using Pi-A isotherm, ellipsometry, and atomic force microscopy (AFM), respectively. When the concentration of PEG lipid was below 1 mol %, the PEG chains could cover the DPPC surface completely in water, but not in the dried state. On the other hand, the PEG chains could cover the phospholipid surface completely in a dried state, as well as in water, when the PEG lipid concentration was above 3 mol %. These PEG chains, showing a brush-type conformation in water, were highly packed and had a bulky structure at the surface in the dried state as well as in the hydrated state. A bulky and extended PEG layer, above 3 mol % concentration, was greatly effective in the prevention of platelet adhesion. PMID- 11033570 TI - Preface. Bioequilavence measures. PMID- 11033569 TI - Effects of geometry of hydroxyapatite as a cell substratum in BMP-induced ectopic bone formation. PMID- 11033571 TI - A practical approach for evaluating population and individual bioequivalence. AB - Pharmacokinetic measurements provided by subjects to each of two formulations of a drug have a joint distribution that can be characterized by parameters reflecting scale and correlation as well as location. The bioavailability of the formulations can be expressed in terms of the means of the marginal distributions, their means and variances, or the marginal means and variances and the joint correlation. These expressions correspond, respectively, to 'average', 'population', and 'individual' bioequivalence when the joint distribution of the measurements is bivariate normal. Current proposals for assessing the degree of bioequivalence of two formulations are based on statistics that are composites of variance components and squares of expected mean differences from a mixed linear model. There are technical and practical issues associated with these proposals, particularly that they require more complicated designs than the familiar 2x2 cross-over. This paper describes an alternative approach that can be applied with standard 2x2 cross-over designs, and that provides evaluations of population and individual bioequivalence that should be adequate for all practical clinical purposes. The approach is based on easily computed correlation and regression coefficients whose statistical properties under normality are well known and for which non-parametric and robust alternatives exist when normality cannot be assumed. The approach yields conclusions consistent with those obtained by the current proposals when applied to data sets supplied by the FDA. In the cases where the conclusions do not match, the new approach appears to be more consistent with the data. PMID- 11033572 TI - The bootstrap procedure in individual bioequivalence. AB - A bootstrap-type hypothesis test procedure for assessing individual (or population) bioequivalence between two drug formulations is suggested in a draft guidance from the United States Food and Drug Administration (FDA). The purpose of this article is to study the unknown properties of this test procedure and propose some improved test procedures. We find that: the FDA's bootstrap computation is not correct; the power of the FDA's test can be very low; the use of the REML method suggested in the draft guidance does not have any advantage over the use of simpler methods such as the moment method; and the method of sample size determination in the draft guidance is inappropriate. We study the size and power of different bootstrap test procedures and suggest a method for sample size determination. It is our hope that this article will draw some attention to further research in this area, and eventually a satisfactory statistical method can be implemented for assessing individual (or population) bioequivalence. PMID- 11033573 TI - On a reasonable disaggregate criterion of population bioequivalence admitting of resampling-free testing procedures. AB - The aggregate criterion of population bioequivalence (PBE) recommended in the new FDA guidance for in vivo bioavailability studies is criticized for using the same distributional parameter as a reference for scaling the squared distance between the averages and the difference in variability. In order to circumvent this and other difficulties inherent in the approach to PBE having been recommended thus far, we introduce a disaggregate criterion instead which requires of population bioequivalent drug formulations that both the difference in means scaled by the pooled intrasubject standard deviation be sufficiently small in absolute value, and the total variability be not substantially increased under the test as compared to the reference formulation. For testing the first of the statistical hypotheses associated with this combined criterion, an exact optimal procedure based on the ordinary two-sample t-statistic is presented. As a test for equivalence with respect to total variability, a modification of Liu and Chow's one-sided test procedure for the assessment of intrasubject variability is recommended. Both subtests are combined by means of the intersection-union principle explained here in general terms, to form an overall test for disaggregate PBE maintaining any specified significance level. The power of the combined procedure is investigated by means of Monte Carlo simulation. PMID- 11033574 TI - The U.S. draft guidance regarding population and individual bioequivalence approaches: comments by a research-based pharmaceutical company. AB - Generally, the motivation for switching from average bioequivalence to population and/or individual bio-equivalence is well recognized in the light of certain limitations of the concept of average bioequivalence. However, this switch still results in unresolved issues which should be addressed before the regulatory guidance is finalized. PMID- 11033575 TI - Numerical methods for the evaluation of individual bioequivalence criteria. AB - The evaluation of individual bioequivalence (IBE) by bootstrap resampling using common statistical software, for example SAS, is extremely time consuming. In this article, an estimation procedure that can be implemented in a high level language with the same degree of accuracy as SAS is described. The necessary parameter estimating equations under both least square (LSE) and restricted maximum likelihood (REML) methods are given. The algorithms used to numerically compute these values are outlined and tested, in FORTRAN, on several simulated data sets and shown to reproduce SAS results with at least 10(-3) precision. More importantly, the REML bootstrap algorithm reduces the time taken in SAS by a factor of 20. Secondary results indicate that LSE and REML parameter estimates are similar for mild unbalancedness. PROC MIXED, with unstructured (UN) and compound symmetry heterogeneous (CSH) variance structures give the same results except when the subject-by-treatment interaction variance, sigma(2)(D), is 0 in which case CSH significantly overestimates sigma(2)(D) and underestimates the within-treatment variances. It is concluded that bootstrap evaluation of IBE is efficiently done using either the LSE or REML algorithm in FORTRAN. PMID- 11033576 TI - Definition of individual bioequivalence: occasion-to-occasion versus mean switchability. AB - Two moment-based scaled definitions of individual bioequivalence are discussed. Based on a mixed effects linear model, their evaluations respectively lead to an unweighted (theta(11)) and a parametric (theta(15)) metric. The two metrics are estimated with respect to study design and two estimation methods. Results show that the two IBE metrics perform equivalently in the fully replicated design. In the semi-replicated design, the definition of theta(11) may not be valid while the evaluation of theta(15) results in a reduction of the weights in the mean difference and switchability components of the metric. Percentage rejection rates in the latter design indicate that theta(11) is more conservative than theta(15). This is because there is an increase of about 15 per cent in the producer risk in theta(11) relative to theta(15) compared to a 7 per cent increase in the consumer risk in theta(15) relative to theta(11). A further disadvantage of the design is that there is a 33 per cent loss in the subject-by-treatment variance efficiency which is reflected in a similar amount of decreased sensitivity to departures from perfect bioequivalence even when more subjects are used to equalize the number of exposure occasions in the two designs. It is concluded that a mean switchability criterion may be more appropriate from an interpretability perspective, the bootstrap resampling method used to evaluate individual bioequivalence based on theta(11) may need to be bias-corrected and that the semi replicated design should be used cautiously. PMID- 11033577 TI - Case studies, practical issues and observations on population and individual bioequivalence. AB - The FDA has proposed replacing the 1992 average bioequivalence (ABE) with population and individual bioequivalence (PBE and IBE). This has led to considerable public discussion between regulatory, academic and industry experts. At the heart of the discussion has been the relatively modest amount of available data to examine the behaviour of the PBE and IBE criteria. A retrospective analysis of 22 data sets from 15 replicate cross-over bioequivalence studies has been conducted (n=12-74). AUC and C(max) parameters from these studies were analysed using ABE, PBE and IBE methods. Of the 22 data sets for AUC, 19 pass ABE, all pass PBE and 20 pass IBE. Of the three data sets that failed ABE, all passed PBE and one passed IBE. The results for C(max) are more variable. Of the 16 data sets where ABE is demonstrated, one data set failed both PBE and IBE. Of the six data sets that failed ABE, two passed both PBE and IBE, three passed PBE but not IBE and one failed all three criteria. There were five data sets that passed ABE and PBE but not IBE. Additional practical issues involving the behaviour of the new criteria and its expected impact on sample size for highly variable drug products will be presented. The characterization of key parameters and their interrelationships will also be discussed with particular emphasis on the subject by formulation term in the IBE criteria. It is concluded that more studies and simulations are desirable before full-scale implementation of PBE and IBE criteria. PMID- 11033578 TI - An individual bioequivalence criterion: regulatory considerations. AB - Over the years, concerns have been raised regarding the appropriateness of using the average bioequivalence approach for evaluation of comparability between formulations. In lieu of average bioequivalence, scientists from academia, industry and regulatory agencies have spent considerable effort and time in exploring the concepts of population and individual bioequivalence, and developing the statistical methods to assess the bioavailability metrics using these approaches. Recently, the Food and Drug Administration (FDA) has published a preliminary draft guidance entitled 'In vivo bioequivalence studies based on population and individual bioequivalence approaches'. The concept of prescribability and switchability underscores the difference between the population and individual bioequivalence approaches. The most important consideration for individual bioequivalence, the focus of this paper, rests on the assurance that products deemed bioequivalent can be used interchangeably in the target population (switchability). In addition to the comparison of averages, the individual bioequivalence approach compares within-subject variabilities and assesses subject-by-formulation interaction. The proposed criterion represents substantial departure from the current practice and thus has resulted in extensive public discussion. In contrast to the current average bioequivalence procedure, the proposed individual bioequivalence approach offers flexible equivalence criteria based on the individual therapeutic window and variability of the reference drug product. The proposed criterion rewards manufacture of less variable drug products, allows scaling criteria for highly variable/narrow therapeutic range drugs, and promotes the use of subjects from the general population in bioequivalence studies. The FDA is currently considering various approaches for resolution of issues raised from the public debate on the subject by-formulation interaction term, statistical methods and resource implications. PMID- 11033579 TI - Connections between average and individual bioequivalence. AB - As pointed out by Anderson and Hauck and others, average bioequivalence does not guarantee that a patient, being on a drug, may switch to a different formulation. The consideration of individual bioequivalence was therefore suggested. We investigate the relationship between these concepts and show how the standard test for average bioequivalence is related to the assessment of individual bioequivalence. This leads to a guide of when this test becomes inappropriate. PMID- 11033580 TI - An alternative index for assessing profile similarity in bioequivalence trials. AB - In a typical bioequivalence trial, summary measures of the plasma concentration versus time profile are used to compare two formulations of a drug product. Commonly used measures include area under the curve (AUC), maximum plasma concentration (C(max)) and time to maximum concentration (T(max)). Equivalence of these summary measures, in general, does not guarantee equivalence of the entire profile. Rescigno and Chinchilli and Elswick propose indices which measure profile similarity, but can be overly sensitive to unimportant differences and are not easily interpreted pharmacologically. We propose an alternative index based on smoothing the relative difference between bioavailability profiles. This provides a method for assessing bioequivalence over the entire profile which has a familiar interpretation and can be tuned to provide a compromise between the insensitivity to pattern differences of summary measures and the oversensitivity of pointwise comparisons. PMID- 11033581 TI - Properties of the estimated variance component for subject-by-formulation interaction in studies of individual bioequivalence. AB - Characteristics of the variance component for the subject-by-formulation interaction (sigma(2)(D)), estimated in simulated studies of individual bioequivalence and in three- and four-period cross-over trials reported by the FDA, were compared. sigma(2)(D) was estimated by (i) restricted maximum likelihood (REML) and (ii) the method of moments (MM). Variation of the variance component, estimated by both procedures (s(2)(D)) and for both the simulated and FDA data, increased with rising intra-individual variation. Consequently, a constant level of s(2)(D) (such as 0.0225 suggested by the FDA) may not be regarded as a basis for demonstrating substantial interactions. Features of the FDA and simulated parameters were similar. The results suggested that the FDA data were compatible with assuming sigma(D)=0.05 or perhaps 0.00. Therefore, there is no foundation for concerns about public health. Both simulations and calculations demonstrated that s(2)(D) estimated by MM was unbiased and its variance was proportional to sigma(4)(WF) when sigma(2)(D)=0. PMID- 11033582 TI - Some statistical considerations on the FDA draft guidance for individual bioequivalence. AB - In December of 1997, the FDA proposed a draft guidance for future in vivo bioequivalence studies. The guidance suggested specific criteria for new drug sponsors to show individual bioequivalence (IBE). The criteria use a mixed scaling aggregate strategy. It has been generally accepted that, under some particular situations, the proposed criteria would result in a relaxation of the current bioequivalence standard set by the average bioequivalence (ABE) criterion. Here we study the magnitude of this relaxation under three scenarios: when the conditions for an ABE investigation are met; when the drugs are highly variable, and when the experiments are poorly conducted. The magnitude of relaxation we report here may be surprisingly large to many. For example, when a drug is highly variable (with the intrasubject coefficient of variation reaching 40 per cent), the allowable limit for the ratio of the formulation means could reach 55-180 per cent in an IBE investigation. In comparison, the usual allowable limit in an ABE investigation is 80-125 per cent. Our investigation raises doubts on whether the implied standard of the new proposed IBE criteria would adequately ensure switchability in highly variable drugs. PMID- 11033583 TI - A small sample confidence interval approach to assess individual bioequivalence. AB - The concept of interchangeable pharmaceutical products has been examined in great detail in the literature. Anderson and Hauck proposed a statistical random coefficient model to study 'switchability', and coined the phrase 'individual bioequivalence' which they defined with a probability-based inequality. Since that paper there has been considerable work and discussion. The Food and Drug Administration has recommended the introduction of individual bioequivalence (IBE) and population bioequivalence (PBE) methods in a draft guidance document. The proposal in the draft guidance includes criteria for IBE and PBE and recommends the use of non-parametric bootstrap 95 per cent upper confidence intervals for the conclusion of either IBE or PBE. However, this method requires intensive computations. We have developed an alternative confidence interval procedure to assess IBE by the FDA recommended criteria. This method utilizes Howe's approximation I to a Cornish-Fisher expansion. Our proposed method is applicable to balanced or unbalanced data in a broad class of extended cross-over designs, and can be easily programmed using readily available software. PMID- 11033584 TI - Cost-effectiveness of colonoscopy in screening for colorectal cancer. AB - BACKGROUND: Fecal occult blood testing, flexible sigmoidoscopy, and colonoscopy are used to screen patients for colorectal cancer. OBJECTIVE: To compare the cost effectiveness of fecal occult blood testing, flexible sigmoidoscopy, and colonoscopy. DESIGN: The cost-effectiveness of the three screening strategies was compared by using computer models of a Markov process. In the model, a hypothetical population of 100 000 persons 50 years of age undergoes annual fecal occult blood testing, sigmoidoscopy every 5 years, or colonoscopy every 10 years. Positive results on fecal occult blood testing or adenomatous polyps found during sigmoidoscopy are worked up by using colonoscopy. After polypectomy, colonoscopy is repeated every 3 years until no polyps are found. DATA SOURCES: Transition rates were estimated from U.S. vital statistics and cancer statistics and from published data on the sensitivity, specificity, and efficacy of various screening techniques. Costs of screening and cancer care were estimated from Medicare reimbursement data. TARGET POPULATION: Persons 50 years of age in the general population. TIME HORIZON: The study population was followed annually until death. PERSPECTIVE: Third-party payer. OUTCOME MEASURE: Incremental cost-effectiveness ratio. RESULTS OF BASE-CASE ANALYSIS: Compared with colonoscopy, annual screening with fecal occult blood testing costs less but saves fewer life-years. A screening strategy based on flexible sigmoidoscopy every 5 or 10 years is less cost-effective than the other two screening methods. RESULTS OF SENSITIVITY ANALYSIS: Screening with fecal occult blood testing is more sensitive to changes in compliance rates, and it becomes easily dominated by colonoscopy under most conditions assuming less than perfect compliance. Other assumptions about the sensitivity and specificity of fecal occult blood testing, screening frequency, efficacy of colonoscopy in preventing cancer, and polyp incidence have a lesser influence on the differences in cost-effectiveness between colonoscopy and fecal occult blood testing. CONCLUSIONS: Colonoscopy represents a cost-effective means of screening for colorectal cancer because it reduces mortality at relatively low incremental costs. Low compliance rates render colonoscopy every 10 years the most cost-effective primary screening strategy for colorectal cancer. PMID- 11033585 TI - Smoking is related to albuminuria and abnormal renal function in nondiabetic persons. AB - BACKGROUND: Smoking induces albuminuria and accelerates progression to renal failure in persons with diabetes, but little is known about the relation between smoking and renal function in nondiabetic persons. OBJECTIVE: To investigate whether smoking is related to albuminuria and abnormal renal function in nondiabetic persons. DESIGN: Cross-sectional study. SETTING: Groningen, The Netherlands. PARTICIPANTS: 7476 participants in the PREVEND (Prevention of REnal and Vascular ENd stage Disease) Study. MEASUREMENTS: Microalbuminuria and high normal albuminuria were defined as urinary albumin excretion of 30 to 300 mg/24 h and 15 to 30 mg/24 h, respectively. Elevated or decreased glomerular filtration rate (GFR) was defined as a creatinine clearance that exceeded or was less than two times the standard deviation of the mean value in nondiabetic, nonsmoking participants who had an albumin excretion of 0 to 15 mg/24 h, adjusted for age and sex. RESULTS: Current smokers had a higher median albumin excretion than nonsmokers and were more likely to have microalbuminuria and high normal albuminuria with elevated or decreased GFR. After adjustment for several potential confounding factors, persons who smoked 20 or fewer cigarettes/d and persons who smoked more than 20 cigarettes/d, respectively, showed a dose dependent association between smoking and high normal albuminuria (relative risk, 1.33 [95% CI, 1.10 to 1.61] and 1.98 [CI, 1.49 to 2.64]), microalbuminuria (relative risk, 1.92 [CI, 1.54 to 2.39] and 2.15 [CI, 1.52 to 3.03]), elevated GFR (relative risk, 1. 82 [CI, 1.31 to 2.53] and 1.84 [CI, 1.12 to 3.02]), and decreased GFR (relative risk, 1.53 [CI, 1.04 to 2.24] and 1.83 [CI, 1.05 to 3. 20]), respectively. Quitting smoking was associated only with microalbuminuria. CONCLUSIONS: Smoking is associated with albuminuria and abnormal renal function. However, these associations are less pronounced or absent in former smokers. PMID- 11033586 TI - Prevalence of type 2 diabetes mellitus among persons with hepatitis C virus infection in the United States. AB - BACKGROUND: Hepatitis C virus (HCV) infection may contribute to the development of diabetes mellitus. This relationship has not been investigated at the population level, and its biological mechanism remains unknown. OBJECTIVE: To examine the prevalence of type 2 diabetes among persons with HCV infection in a representative sample of the general adult population of the United States. DESIGN: Cross-sectional national survey. SETTING: The Third National Health and Nutrition Examination Survey, 1988-1994. PARTICIPANTS: 9841 persons older than 20 years of age for whom data on HCV infection and diabetes were complete. MEASUREMENTS: The presence of diabetes was ascertained by using American Diabetes Association guidelines based on fasting plasma glucose measurement and medication history. Presence of HCV infection was assessed by testing for serum HCV-specific antibodies (anti-HCV). RESULTS: Of the 9841 persons evaluated, 8.4% had type 2 diabetes and 2.1% were anti-HCV positive. Type 2 diabetes occurred more often in persons who were older, were nonwhite, had a high body mass index, and had low socioeconomic status. Type 2 diabetes was less common in persons who acknowledged previous illicit drug use. After adjustment for these factors, persons 40 years of age or older with HCV infection were more than three times more likely than those without HCV infection to have type 2 diabetes (adjusted odds ratio, 3.77 [95% CI, 1.80 to 7.87]). None of the 19 persons with type 1 diabetes were anti HCV positive. CONCLUSION: In the United States, type 2 diabetes occurs more often in persons with HCV infection who are older than 40 years of age. PMID- 11033587 TI - Long-term outcomes of scleroderma renal crisis. AB - BACKGROUND: Although scleroderma renal crisis, a complication of systemic sclerosis, can be treated with angiotensin-converting enzyme (ACE) inhibitors, its long-term outcomes are not known. OBJECTIVE: To determine outcomes, natural history, and risk factors in patients with systemic sclerosis and scleroderma renal crisis. DESIGN: Prospective observational cohort study. SETTING: University program specializing in scleroderma. PATIENTS: 145 patients with scleroderma renal crisis who received ACE inhibitors and 662 patients with scleroderma who did not have renal crisis. MEASUREMENTS: Among patients with renal crisis, the four outcomes studied were no dialysis, temporary dialysis, permanent dialysis, and early death. Demographic, clinical, and laboratory data were compared to identify risk factors for specific outcomes. Follow-up was 5 to 10 years. RESULTS: 61% of patients with renal crisis had good outcomes (55 received no dialysis, and 34 received temporary dialysis); only 4 of these (4%) progressed to chronic renal failure and permanent dialysis. More than half of the patients who initially required dialysis could discontinue it 3 to 18 months later. Survival of patients in the good outcome group was similar to that of patients with diffuse scleroderma who did not have renal crisis. Some patients (39%) had bad outcomes (permanent dialysis or early death). CONCLUSIONS: Renal crisis can be effectively managed when hypertension is aggressively controlled with ACE inhibitors. Patients should continue taking ACE inhibitors even after beginning dialysis in hopes of discontinuing dialysis. PMID- 11033589 TI - Making the Rounds. PMID- 11033588 TI - Diagnosis and management of infections involving implantable electrophysiologic cardiac devices. AB - BACKGROUND: Optimal treatment of infections related to implantable electrophysiologic cardiac devices is poorly defined. OBJECTIVE: To describe the clinical presentation, treatment, and outcome of patients with such infections. DESIGN: Retrospective case series. SETTING: The Cleveland Clinic Foundation, Cleveland, Ohio. PATIENTS: 123 patients with infections involving implantable cardiac electrophysiologic devices. MEASUREMENTS: Demographic characteristics, clinical manifestations, time to diagnosis, management, and outcome. RESULTS: 87 patients with permanent pacemakers and 36 patients with implantable cardioverter defibrillators had infections. The most common signs and symptoms were pocket erythema and local pain. The most common pathogens were coagulase-negative staphylococci (68%) and Staphylococcus aureus (23%). In 117 patients (95%), all equipment was extracted and antibiotic therapy lasted a median of 28 days. Operative mortality was zero. Follow-up showed crude mortality and relapse rates of 8% and 3%, respectively. CONCLUSION: For infections related to implantable electrophysiologic devices, complete device removal and antimicrobial therapy allow timely, successful reimplantation at a remote anatomic site without substantial risk for operative mortality or recurrent infection. PMID- 11033590 TI - Better access to information about clinical trials. AB - Access to information about clinical trials is important to researchers, health care professionals, and patients. Many have argued for the establishment of clinical trials registries, citing their substantial benefits. Although some registries do exist, it has been difficult to create comprehensive, easily accessible systems. This paper briefly reviews existing registries, discusses the challenges in building registries, and reviews some of their benefits. The paper concludes with a description of a new, extensive Web-based registry called ClinicalTrials.gov (http://clinicaltrials. gov/), which was developed at the National Institutes of Health (NIH) by the National Library of Medicine as a result of recent legislation calling for a comprehensive, publicly accessible registry of clinical trials. The first version of the system became available in late February 2000 and contains information about approximately 5000 trials. The first release contains primarily NIH-sponsored trials, and new trials are regularly added to the system. Subsequent versions will contain information about trials sponsored by other federal agencies and by the private sector. The system was developed in accordance with basic informatics principles, including adherence to standards, usability considerations, and iterative testing and evaluation. PMID- 11033591 TI - Update in oncology. PMID- 11033592 TI - Cachexia. AB - Cachexia represents the clinical consequence of a chronic, systemic inflammatory response, and its manifestations differ considerably from those of starvation. Although cachexia is classically associated with chronic infections and malignant conditions, some of its elements have been identified in a wide variety of chronic diseases and in aging persons. Cachexia has repeatedly been associated with adverse clinical outcomes. The changes seen in cachexia are multidimensional and highly coordinated. Most obvious is a redistribution of the body's protein content, with preferential depletion of skeletal muscle and an increase in the synthesis of proteins involved in the response to tissue injury-the so-called acute-phase response. The physiologic, metabolic, and behavioral changes of cachexia are tightly regulated by cytokines, which signal the synthesis of acute phase proteins as well as changes in intermediary metabolism that provide substrate and energy. The metabolic adaptations, notably the increase in the rate of protein degradation, limit the ability of hypercaloric feeding to reverse the depletion of lean mass. Recent studies have demonstrated the ability of anabolic and anticatabolic agents to mitigate the loss of skeletal muscle and to improve clinical outcomes in selected circumstances. Preclinical initiatives target the cytokine regulation of protein metabolism. It should be stressed that metabolic manipulation in cachexia could have positive or negative clinical effects, which must be distinguished through appropriate clinical trials. PMID- 11033593 TI - Osteoarthritis: new insights. Part 1: the disease and its risk factors. AB - Osteoarthritis is the most common form of arthritis, affecting millions of people in the United States. It is a complex disease whose etiology bridges biomechanics and biochemistry. Evidence is growing for the role of systemic factors (such as genetics, dietary intake, estrogen use, and bone density) and of local biomechanical factors (such as muscle weakness, obesity, and joint laxity). These risk factors are particularly important in weight-bearing joints, and modifying them may present opportunities for prevention of osteoarthritis-related pain and disability. Major advances in management to reduce pain and disability are yielding a panoply of available treatments ranging from nutriceuticals to chondrocyte transplantation, new oral anti-inflammatory medications, and health education. This article is part 1 of a two-part summary of a National Institutes of Health conference. The conference brought together experts on osteoarthritis from diverse backgrounds and provided a multidisciplinary and comprehensive summary of recent advances in the prevention of osteoarthritis onset, progression, and disability. Part 1 focuses on a new understanding of what osteoarthritis is and on risk factors that predispose to disease occurrence. It concludes with a discussion of the impact of osteoarthritis on disability. PMID- 11033594 TI - Prevention and treatment of colorectal cancer: pay now or pay later. PMID- 11033595 TI - An association between hepatitis C virus infection and type 2 diabetes mellitus: what is the connection? PMID- 11033596 TI - Magic in the Web of it. PMID- 11033597 TI - Winter Desire. PMID- 11033598 TI - To Nowhere. PMID- 11033599 TI - Drug effects on driving performance. PMID- 11033600 TI - Drug effects on driving performance. PMID- 11033601 TI - Drug effects on driving performance. PMID- 11033602 TI - Drug Effects on Driving Performance. PMID- 11033603 TI - Rosiglitazone toxicity. PMID- 11033605 TI - Correction: Walnuts and Serum Lipid Profile. PMID- 11033604 TI - Over-the-counter availability of H2-receptor antagonists and gastric cancer. PMID- 11033606 TI - Correction: Calcific Constrictive Pericarditis. PMID- 11033608 TI - Therapy of Digestive Disorders: A Companion to Sleisenger and Fortran's Gastroenterological and Liver Disease. PMID- 11033607 TI - Hormone Therapy and the Brain: A Clinical Perspective on the Role of Estrogen. PMID- 11033609 TI - Vaccines today. PMID- 11033610 TI - HIV testing during pregnancy. Building the case for voluntary testing. PMID- 11033611 TI - FDA proposes new rules regarding trans fatty acids. PMID- 11033612 TI - Update on breast and cervical cancers. PMID- 11033613 TI - Exploring dental hygiene and perinatal outcomes. Oral health implications for pregnancy and early childhood. PMID- 11033614 TI - Netiquette. Maintaining confidentiality and privacy on discussion lists. PMID- 11033615 TI - Making mistakes. Prevention is key to error-free health care. PMID- 11033616 TI - Beyond the law. One nurse's reflections on conflicts and consequences in mandated reporting. PMID- 11033617 TI - Solution-focused therapy. Is it useful for nurses in the workplace? PMID- 11033618 TI - A new millennium in nursing care. AWHONN prepares for 2000 and beyond. PMID- 11033619 TI - Learning from those we serve. PMID- 11033620 TI - Consultant nurse role may divide the profession. PMID- 11033621 TI - Dr Shipman: how could it have been prevented? PMID- 11033622 TI - Case 20: improper seclusion. Improperly securing patients in their rooms or in a toilet. PMID- 11033623 TI - Blood transfusion: crucial steps in maintaining safe practice. AB - The transfusion of blood and blood products remains a highly effective and potentially life-saving treatment for many patients. However, blood is a living tissue and its transfusion, from one individual to another, is not without risk. Significant among these risks is the potential for human error and the subsequent transfusion of the incorrect blood component. Alarmingly, the number of errors would appear to be increasing, with the second annual report of the Serious Hazards of Transfusion (SHOT) Steering Group reporting a 36% increase in these cases (SHOT, 1999). Reversing this trend can only be achieved by adopting all inclusive and rigorous blood transfusion procedures. The registered nurse is central to such processes and therefore plays a crucial role in the safe administration of blood and blood products. This article considers the nurse's responsibilities and offers evidence-based guidelines for practice. PMID- 11033624 TI - Avoiding problems: evidence-based selection of medical gloves. AB - Allergenic contact reaction to glove proteins and residual chemicals increased from 1980 onwards; circumstantial and technical evidence indicates that change from reusable to disposable latex gloves was a causative factor. Several protein fractions are potentially allergenic; if low protein gloves contain a dominant protein fraction, people with sensitivity may still have an adverse reaction. Use of chlorinated powder-free gloves reduces risk of allergenic reaction; other powder-free gloves may still pose a contact risk to patients. Adverse comparative tests of vinyl vs latex medical gloves may, in some cases, have used non-medical gloves without assessment of initial quality or involved an unrealistic test method; these have discouraged use of a non-allergenic alternative to latex gloves. Vinyl medical glove properties are reviewed. Nitrile examination gloves offer better protection than latex types when handling lipid-soluble substances and chemicals. Properties of medical gloves made from other materials are described. PMID- 11033625 TI - Confidentiality. 13: The notification of infectious diseases. AB - Jenny Rose was a paediatric community nurse who regularly visited a child with a chronic lung condition who was being nursed at home. On one visit she noticed that the child's mother, Jane, appeared to be very pale and thin and was told that the mother had a severe gastric disorder with diarrhoea. From the description of the illness, Jenny thought that Jane might be suffering from typhoid. Jane worked as a cook in a restaurant, was unwilling to seek medical advice and intended going to work that night. Jenny was concerned that Jane could have a serious notifiable infectious disease and therefore be a danger to customers in the restaurant. Jane insisted that Jenny should keep the information confidential. Where does Jenny stand? PMID- 11033626 TI - A patient temperature audit within a theatre recovery unit. AB - Hypothermia in the postoperative recovery phase can be a source of potential danger for patients emerging from anaesthesia. In spite of the vast amount of literature dealing with principles of care in the recovery room, information regarding the incidence of hypothermia appear very limited. This article presents a patient temperature audit to assess whether patients were being discharged from a theatre recovery unit hypothermic. In addition, patient temperatures were recorded both pre- and postoperatively, as well as on discharge from recovery, to monitor the incidence of hypothermia throughout the theatre suite. The results show that patients were being discharged with core body temperatures ranging from 34.8 to 38 degrees C with no incidence of hypothermia recorded when the operating room temperature was above 23 degrees C. In general, the coldest theatres were orthopaedic, with this group of patients showing the most incidence of hypothermia. This article aims to demonstrate the use of clinical audit to investigate and evaluate current practice. PMID- 11033627 TI - Gerontological nursing. 3: Valuing nursing homes and valuing staff. AB - This, the third of five articles considering the future of gerontological nursing, focuses on practice within nursing homes. The important contribution of such environments is highlighted and it is argued that work within long-term care settings must be recognized and valued if the quality of care provided is to improve. PMID- 11033628 TI - Web watch. PMID- 11033629 TI - Infection control: reducing the psychological effects of isolation. AB - Although previous studies have shown that some patients, when nursed in isolation for infection control purposes, experience negative emotional effects, little research has been published which suggests what healthcare professionals can do to prevent or reduce these effects. A study was therefore carried out involving 21 patients being nursed in isolation who completed questionnaires. Five patients with negative experiences of isolation were then interviewed about the reasons for feeling the way they did and what they felt could be done to reduce or prevent negative emotions. Recommendations made from the research include the provision of written and individualized information, improved communication from medical staff, provision of a common room and better facilities to relieve boredom. Further large scale research is needed to investigate strategies to improve the isolation experience. PMID- 11033630 TI - Clinical supervision in mental health nursing courses. AB - The Department of Health (DoH, 1994) advocated the introduction of clinical supervision into mental health nursing practice and suggested that student nurses be prepared in what to expect from this process. The ENB (1995) supported this recommendation but has offered no guidelines on how it is meant to be implemented. This article reports on an educational initiative in which group supervision was implemented within one cohort of preregistration mental health nursing students. The students reported a number of perceived benefits: a greater understanding of the purpose and benefits of clinical supervision; skill development; the opportunity to reflect on practice; and the reduction of stress. PMID- 11033631 TI - Partnership is the key to nursing's future. PMID- 11033632 TI - The amputation of healthy limbs is not an option. PMID- 11033633 TI - Florence Nightingale must remain as nursing's icon. PMID- 11033634 TI - Case 21: unsafe environment. Caring for elderly patients in a substandard environment. PMID- 11033635 TI - Malnutrition and pressure ulcers: nutritional assessment tools. AB - Good nutrition is essential throughout life and often patients have a poor understanding of what a balanced diet comprises. Nurses need to have a good comprehension of malnutrition. Where malnutrition is evident it can be resolved by working with the patient to encourage a healthy diet. One way to improve patient nutrition is by a nutrition audit of patient food. Numerous nutritional assessment tools exist which help to identify a patient's nutritional status. No nationally standardized nutritional assessment tool exists and there is a danger of assessment methods becoming as proliferate as pressure ulcer risk assessment tools. A solution to this problem may be to use a pressure ulcer risk assessment tool with a nutritional risk assessment tool such as the Burton Score. Pressure ulcers and malnutrition have been demonstrated to be strongly correlated with each other. Malnutrition is not just present in hospitals but also in nursing homes. A solution to this problem is to use nutritional tools to highlight nutritional deficiency in a patient's diet not only in hospitals but also in the community and in nursing homes. PMID- 11033636 TI - Gerontological nursing. 4: Age-related hearing explored. AB - In the previous three articles in this series (Vol 9(1): 39-42; Vol 9(2): 103-6; Vol 9(3): 157-60), nurses have been challenged to reconsider their approach towards the care of older people. To facilitate this it is helpful to provide specific examples of how expert practice might be achieved. Using one of the most prevalent but neglected problems of old age, hearing disability, the authors describe how nurses can contribute to the health of older people and maximize the therapeutic component within gerontological nursing. PMID- 11033637 TI - Systematic approach to community risk assessment and management. AB - A move from institutional to community health care means that health service staff are increasingly requested to visit patients in their own homes. This undertaking is not without risk, particularly where the patient or the locality is unknown. There are no nationally available guidelines for formally assessing potential risk to a health worker before the home visit. A protocol for risk assessment and a safety schedule before making a home visit was therefore developed and is described in this article. The difficulties in carrying out a comprehensive risk assessment are outlined. Several ways in which the assessment of risk before home visits could be made more effective are suggested. PMID- 11033638 TI - Attendance for self-harm in a West Midlands hospital A&E department. AB - This article focuses on the issue of self-harm. It reviews the literature and details specifically a comparison of data collected in the accident and emergency (A&E) department of a hospital in the West Midlands in 1990 and in 1997. Both the research literature and the data collected at the hospital suggest that there is a rise in the numbers of people self-harming and presenting to A&E departments. Any changes in rates of self-harm or the demographic characteristics of the patient population are likely to have important implications for clinical services. Additionally, the links between self-harm and suicide are well documented (Hawton and Fagg, 1988; Morgan and Williams, 1994) and there is an urgent need to ensure quality service for this high-risk group. An effective assessment of the area is difficult. The limited number of comprehensive studies prohibits definitive epidemiological data. Furthermore, confusion over terminology hinders clear analysis and comparison of that which is collected. PMID- 11033639 TI - How nurses can build trusting relationships with clients. PMID- 11033640 TI - Web Watch. PMID- 11033641 TI - Antibiotic resistance: a growing and multifaceted problem. AB - Antibiotic resistance is an increasing problem worldwide that is exacerbated by the overuse and misuse of antibiotics. Patients, pharmaceutical marketing, and the use of antibiotics in veterinary medicine and animal husbandry are important factors to consider in the emergence of resistance. Infection control measures to prevent the spread of antibiotic-resistant bacteria are compromised by poor compliance to basic measures such as handwashing and standards of environmental cleanliness. Wider epidemiological factors such as global travel and complacency towards public health must also be considered. This article aims to improve understanding of antibiotic resistance and suggests ways in which nurses can contribute towards the strategy to address the problem. PMID- 11033643 TI - The Verge Videometer wound measurement package. AB - Wound measurement can be made as simple or as complex as time allows, but there is no doubt that wound measurement is required in some degree as part of the patient documentation process. To help assess the progress of wound healing, BES Rehab Ltd is distributing the Verge Videometer (VeV), a wound tracking system which can be used for precise wound measurement, tracking and documentation. The VeV is suitable for use in hospitals, outpatient clinics and in the community as well as being useful for those conducting research into wound progression. PMID- 11033642 TI - Safe use of medical gases in the clinical setting: practical tips. AB - For a variety of reasons, nurses, midwives and health visitors may have to be involved in administration of many types of medical gases. In order to provide safe and effective care the nurse must be aware of the implications of his/her practice. This article discusses and describes the many types of medical gases available in clinical practice. The article focuses on four commonly used gases: oxygen, air, nitrous oxide and entonox. Practical tips are offered and the focus is made on the safe and effective use of the gases. Details regarding the storage and administration of medical gases are outlined. Finally, some of the potential dangers associated with the gases are discussed. PMID- 11033644 TI - A nursing career in the Army. PMID- 11033645 TI - Nursing needs defining: new initiative launched. PMID- 11033646 TI - The way forward. PMID- 11033647 TI - Naturopathy, Nightingale, and nature cure: a convergence of interests. AB - Naturopathy is making a strong resurgence in many Western countries, and its philosophical and theoretical basis has much in common with nursing theories of health. This article will give an overview of naturopathy, its origins in Hippocratic medicine and nature cure, and its re-emergence during the Victorian era in the context of a renewal of interest in health. The question is posed, 'To what extent did the nature cure movement of last century influence the thinking of Florence Nightingale?' An examination of her writings supports the hypothesis that nature cure was a significant influence on Nightingale's understanding of health and healing. Contemporary nursing theory is also congruent with naturopathic principles in significant ways, and a convergence between the two disciplines is emerging. PMID- 11033648 TI - Healthcare: integrated in the fabric of life. AB - Healing as a primary function of life touches all aspects of an individual as well as of the collective, be that a community or our society as a whole. Living for the last II years in an intentional community, the Findhorn Community in north-east Scotland, the author has the privilege to explore the relevance of healthcare for individuals and as a community service. Healthcare is much more than medical care. Mostly it is a basic aspect of daily life and relies on self responsibility, self-care and appropriate resources/opportunities. Should medical care become necessary a wide choice of treatment approaches offered to the patient facilitates patient empowerment and involvement. The author, therefore, advocates the integration of orthodox and complementary medicine. PMID- 11033649 TI - CAM research and development. AB - The growth in complementary medicine and its integration into conventional medicine will certainly require the development of a well defined research strategy within the UK. This paper summarizes discussions that occurred under the umbrella of the Foundation for Integrated Medicine, involving both conventional and complementary medical practitioners. It is the first UK-based consensus document that defines a research strategy for complementary and alternative medicine and looks specifically at problems such as priority setting, research methodology, research capacity and support, potential funding streams and possible routes for the dissemination of CAM research. It concludes that for CAM to have an established future within UK healthcare, it must be subject to rigorous scrutiny. The methods employed should be appropriate to the task and should embrace a wide spectrum, both quantitative, qualitative and incorporating projects that involve good practice and audit. PMID- 11033650 TI - Effect of cutaneous stimulation on pain reduction in emergency department patients. AB - CONTEXT: The problem of unrelieved pain in Emergency Department (ED) patients as well as contraindications to analgesics and narcotics due to masking of symptoms during the diagnostic period, presents itself as a unique opportunity for ED nurses to utilize non-pharmacological and non-invasive interventions such as cutaneous stimulation (CS) to relieve pain. OBJECTIVE: To evaluate the effectiveness of a specific protocol of CS, developed by the researchers, in reducing pain levels in ED patients. Another objective was to determine the effect of CS on blood pressure and heart rate. Potential factors that could influence the dependent variables such as age, gender, educational level, location of pain, and site of CS were tested. DESIGN: A one group pre-test post test experimental design measured variables before and after intervention in all subjects. SAMPLE: After being screened for inclusion, 50 patients (38 adults, 12 children) were admitted to the study and were treated with CS to relieve pain. RESULTS: Following CS, subjects reported significantly reduced pain, and demonstrated reduced heart rate, and blood pressure readings. The location of pain significancy influenced heart rate and diastolic blood pressure but not pain level. The most effective site of CS was contralateral to the pain. Age, gender and educational level had no significant affect. CONCLUSIONS: The results of this study provide empirical evidence that CS effectively reduces pain, heart rate, and blood pressure in ED patients. The intervention of CS has solid utilization potential and could be easily incorporated into standard ED procedure. Further research is needed to identify threats to internal validity especially that of the caring presence of the nurse. PMID- 11033651 TI - The use of aromatherapy in intrapartum midwifery practice an observational study. AB - The authors report the process and results of an evaluation of a midwifery aromatherapy service for mothers in labour: This study of 8058 mothers in childbirth, is the largest research initiative in the use of aromatherapy within a health-care setting. The study involved a wide range of participants, from mothers who experienced a low risk, spontaneous labour and birth, to those whose labour was induced, and those who had vaginal operative delivery and Caesarean section. The study-took place over a period of 8 years, which enabled a more challenging test of the effect of aromatherapy on intrapartum midwifery practice and outcomes. In the study a total of 10 essential oils were used, plus a carrier oil, which were administered to the participants via skin absorption and inhalation. The study found little direct evidence that the practice of aromatherapy per se reduces the need for pain relief during labour, or the incidence of operative delivery. But a key finding of this study suggests that two essential oils, clary sage and chamomile are effective in alleviating pain. The evidence from this study suggests that aromatherapy can be effective in reducing maternal anxiety, fear and/or pain during labour. The use of aromatherapy appeared to facilitate a further reduction in the use of systemic opioids in the study centre, from 6% in 1990 to 0.4% in 1997 (per woman). Aromatherapy is an inexpensive care option. In 1997 when 1592 mothers used aromatherapy, the total cost was 769.17 Pounds. The study reports a minimal incidence of associated symptoms. Out of 8058 mothers, 1% (100) recorded an associated symptom. These were mild in nature. The successful model of integrated practice that this aromatherapy study presents, offers a useful example for other units to consider. PMID- 11033652 TI - Healing relationships with nature. AB - Our health is intimately connected to the health of our environment. The contemporary world view which sees a radical distinction between humans as subjects and world as object can obscure our recognition of how much we rely on nature for health and survival. Indigenous traditions and contemporary scholars remind us that we live in a universe in which all things are connected, and in which nature continues to offer its gifts in co-creative partnership for the health and wellbeing of all. Living in awareness of our relationship with nature enables us to open more to the experience of nature's nurturing. Many complementary therapies derive from ancient practices that involve nature in healing partnership. As nurses and midwives we must learn to expand our boundaries to encompass the many ways that nature partners with us for healing. PMID- 11033653 TI - Complementary medicine: knowledge, use and attitudes of doctors. AB - The recent growth in interest in the use of complementary medicine by the general public has been well reported. Health professionals, particularly nurses and midwives, are incorporating the use of complementary therapies into their practice. The medical profession is involved in the political processes affecting legislation governing complementary medicine and it is therefore important to determine the views of doctors when considering the use of complementary therapies by nurses within the health-care system. A critical review of published research studies which consider the knowledge, use and attitudes of doctors to complementary medicine is presented. These studies suggest doctors' interest in complementary medicine but they also raise a number of concerns including lack of evidence to demonstrate effectiveness, possible harmful effects, inadequate knowledge of doctors and lack of statutory regulation for most therapies. Recommendations of recent reports are outlined which attempt to address these and other issues. PMID- 11033654 TI - Biocidal Products Directive 98/8/EC. PMID- 11033655 TI - Policy development. AB - The implementation of certain therapies within clinical practice is dependent on nurses taking the initiative and expanding their scope of practice. In incorporating therapies within nursing practice it is hoped that a more holistic approach to care can be achieved. PMID- 11033656 TI - Cost, quality, and satisfaction with hospice after-hours care. AB - Three hundred seventy-three patients enrolled in a health maintenance organization's hospice program were given two different models of after-hours nursing care, one using nurse employees and the other using contract nurses. Statistically significant differences were found between the two groups in cost, in quality of nursing care, and in patient/family satisfaction. Cost of hospital days for the organization was less when patients received after-hours care from nurse employees of the organization, and quality of documentation was better. More patients who received care from nurse employees were satisfied with nurses' arrival time and understanding of problems as well as with the service, information, courtesy, follow-up, and respect which patients received during their hospice stay. PMID- 11033657 TI - The role of primary caregiver denial in inpatient placement during home hospice care. AB - This study conducted with a sample of 68 home hospice patients revealed that primary caregivers in denial of the patient's terminality were more likely to place hospice patients in inpatient treatment. Moreover, patients placed in inpatient settings were more likely to die there, rather than at home as planned. These findings suggest an impact of primary caregiver denial upon patient self determination, and indicate the importance of addressing denial in counseling with primary caregivers of terminally ill patients. PMID- 11033659 TI - Nurses' work in a hospice and in an oncological unit in Sweden. AB - This paper presents the results of a work-sampling study aimed at describing and comparing the activities of nurses at a free-standing in-patient hospice and a hospital oncological unit. Data suggests that although patient care at both sites is structured by organizational routines, the content and quantity of the nurses' physical and emotional labor differed markedly. Hospice nurses spend significantly more of their working time with patients and/or relatives (37%) than the oncological nurses (21%). When they are with patients the nurses in both settings are most often performing a physical activity such as helping patients with their daily activities in the hospice, and helping patients with needs in relation to investigations and treatment in the oncological unit. PMID- 11033658 TI - The evolution of volunteerism and professional staff within hospice care in North Carolina. AB - Although the pursuit of Medicare certification has been one of the most controversial events in hospice history no study has examined its impact using data from a defined population of hospices before, during, and after federal legislation was enacted. This paper revisits the debate over the changing role of volunteers using such a longitudinal data source. Over time, as the patient population grew, aggregate levels of volunteers and professional staff increased. However, the ratios of professional staff and volunteers to patients reveal that regardless of certification status, hospices retained more professional staff per patient and fewer volunteers per patient over time. These data suggest hospices, particularly certified organizations, have transitioned from voluntary organizations to professionally staffed organizations with a strong volunteer component. The most important issue for future research is whether the observed changes have affected the quality of hospice care. PMID- 11033660 TI - Matching bereavement services to level of need. AB - The establishment of hospices in the U.S. has resulted in an increase in bereavement services available to clients experiencing loss. Many of these services, such as educational and peer support groups, are provided without screening or assessment of risk for complicated bereavement in the individuals who utilize them. While acceptance of uncomplicated bereavement is important, assessment for complicated bereavement is also important to ensure that appropriate services are offered to those at risk for problematic adjustment. This article reviews the literature on risk criteria for complicated bereavement and proposes a method both for assessing risk and matching service to level of need, based on these criteria. PMID- 11033661 TI - Where has all the compassion gone? PMID- 11033662 TI - Ophthalmic support staff salaries: skills, certification, practice setting are all factors. PMID- 11033663 TI - Improved technology makes autorefractors a fundamental part of today's busy office. PMID- 11033664 TI - Ocular risks from sunlight exposure. AB - Relinquishing the desire to acquire a tan will be difficult for many people. To many individuals, a tan is a symbol of health, fashion and attractiveness. Even when informed about the risks of sunlight exposure, many people ignore the advice to change their behaviors and decrease their risks. Adoption of safe sunlight precautions depends on individual preference, age, work requirements, gender, and perception of personal risk. Many people selectively adhere to certain protective behaviors which do not conflict with social norms. Reinforcement of how sunlight can damage the eyes and skin needs to be continually provided so that public attitudes can adapt to all the necessary appropriate behaviors. Ophthalmic health care professionals have an important part in public education. They can customize their patient's personal protective requirements to match the type of risks uncovered in the clinical work up. The ocular and social history should include the patient's sunlight exposure risk profile and document pre-existing sunlight damage to the eyes and facial areas. All follow-up visits should reinforce sunlight exposure information and document compliance or non-compliance. Only in this way can the eye care delivery system support preventative measures to decrease ocular injuries from sunlight radiation. PMID- 11033665 TI - Quality and cost effectiveness in health care: a unique approach. PMID- 11033666 TI - Pharmacologic protocol varies for LASIK. PMID- 11033667 TI - Hyperopic refractive surgery solutions for the new millennium. PMID- 11033668 TI - Is seeing believing? Visual hallucinations in age-related macular degeneration and Charles Bonnet. PMID- 11033669 TI - Disposable and planned replacement lenses for the new millennium. PMID- 11033670 TI - Measuring the outcomes of school nursing practice: showing that school nurses do make a difference. PMID- 11033671 TI - Screening adolescents with mental retardation for depression. AB - This study investigated the reliability and validity of the Children's Depression Inventory (CDI) using 27 adolescents with mild mental retardation attending schools in a large metropolitan area. Cronbach's alpha was 0.79 and the test/retest correlation was 0.90 at 10 days to 2 weeks, indicating adequate reliability. Scores on the self-report CDI were compared with scores on an observer-completed inventory of psychopathology filled out by teachers and other staff, the Reiss Scales for Children's Dual Diagnosis. For female students, statistically significant negative associations were found between scores on the CDI and scores on the Reiss Scales and its depression subscale. Screening adolescents with mental retardation for depression should be an important part of Individualized Educational Plans for school nurses. School staff report should not be the only method of screening for depression in adolescents with mild mental retardation. School nurses may find the CDI useful as a self-report screening tool for depression. PMID- 11033672 TI - A coordinated school health program approach to adolescent obesity. AB - Obesity has become one of this country's most significant nutritional diseases. The prevalence of childhood and adolescent obesity has steadily increased over the past 20 years. The risk of developing health problems increases as the obese child becomes an obese adolescent and adult. The 1997 Youth Risk Behavior Survey reports adolescents participate in behaviors that put them at risk for obesity. The recognition of these unhealthy behaviors among our youth has led to the need for early intervention. The Coordinated School Health Program is a mechanism to address adolescent obesity at the school-age level. This program includes an organized set of policies, procedures, and activities intended to protect and promote the health and well-being of students and staff. The eight components of a Coordinated School Health Program (CSHP) include school health services, a healthy school environment, comprehensive school health education, counseling and guidance, physical education, food service, worksite health promotion, and the integration of school and community. This paper describes integration of prevention and treatment strategies for adolescent obesity for each of the eight components of a CSHP. PMID- 11033673 TI - Empowering students on medication for asthma to be active participants in their care: an exploratory study. AB - This article is part of a larger study of young students with asthma. It focuses on the observations of school nurses and comments from parents and students pertinent to school nursing practice. The study shed light on the educational needs of 9- to 11-year-old students with asthma and identified some areas where the school nurse could easily enhance the health management of these students. Responses from students and parents about asthma provided insight into the daily challenges faced by students with asthma. Students with asthma may or may not fully understand the seriousness of their chronic condition or how to recognize and treat symptoms. The purpose of this pilot study was to identify strategies school nurses might use to determine the knowledge level of students and to determine the effects of asthma education programs. School nurses who participated in the study were provided with a comprehensive package containing current asthma information and an education program to update knowledge and skills. These school nurses also received Open Airways for Schools, an American Lung Association education program for students in grades three to five. The study was a two-group exploratory, quasi-experimental design. Questionnaires were prepared for students, school nurses, and parents to collect information on knowledge and perceptions about asthma. Students who participated in the educational program were reported by their school nurses to have increased knowledge and management skills related to their disease. PMID- 11033674 TI - A World Wide Web-based health resource. Survey of Missouri school nurses to determine priority health information resources for SchoolhealthLink. AB - Two hundred ninety-two school nurses in Missouri participated in a mailed survey to aid in the design of SchoolhealthLink, a World Wide Web (WWW)-based information service for Missouri school nurses and children. The nurses identified specific health information resources likely to benefit school nurses and school children and prioritized these resources. The school nurses assigned high priority to 11 types of health information resources: individualized health care plans, emergency care plans, communicable disease control plans, acute illness, injuries, communicable diseases, hotline numbers, medications in schools, immunization protocols and standards, community-based health care resources, and Department of Elementary and Secondary Education regulations that affect school nurse practice. The four most common health problems school nurses identified were asthma, attention deficit disorder, diabetes, and head lice. SchoolhealthLink will provide a one-stop WWW-based resource for school nurses and school children. PMID- 11033675 TI - Creating a living document: the National Association of School Nurses Strategic Plan 1998-2001 mid-course evaluation. AB - This is the third in a series of three articles devoted to the development, implementation, and evaluation of the National Association of School Nurses 1998 2001 Strategic Plan and how it relates to the practice of school nursing. Information regarding the extent to which each of the seven goals in the Strategic Plan has been met, and the future efforts in developing the 2001-2004 Strategic Plan will be reviewed. Areas of focus of the new NASN leaders and the priorities of the NASN management and Board of Directors will shape the future goals of the organization. The new definition of the practice of school nursing, the NASN Mission Statement, and the current Strategic Plan goals will drive the development of the next Strategic Plan. NASN leaders have a firm grasp on the future direction of the organization and are available and responsive to assist individual members and state affiliates in managing change and empowering school nurses to meet the challenges of the future. PMID- 11033676 TI - Identifying high school students' health behaviors: development of a survey. AB - A health survey developed to profile high school students for the purpose of assisting in program planning emphasized everyday health behaviors instead of focusing on high-risk behaviors (e.g., drinking, drugs, safety, or sexuality). The questionnaire was a voluntary 20-item survey that could be completed during a 15-minute homeroom period. A total of 1,401 students completed the questionnaire. Findings were shared with students, teachers, staff, and parents. Teachers and staff members were given a prepared response sheet that addressed each of the concerns identified. They were encouraged to have discussions about these behaviors with students during the homeroom period, as time and interest allowed. Response to the survey has been very positive, and a number of changes have been incorporated into the school health program as a result of the findings. One of these changes is a teen clinic that opened in April 1999. The findings also proved very useful in facilitating awareness of student behaviors and assisted in identifying ways the school nurse could assist students in improving their health and well-being. PMID- 11033677 TI - Nurse practitioner student prescriptive patterns. AB - As employment of nurse practitioners (NPs) increases in health care systems, there is a need to have current data on their prescribing practices and patterns, and to implement a system for updating such data. This study reports prescriptive data based upon 10,421 primary care visits conducted by 55 family NP students over a 15-month period in 1997 and 1998. Numbers of over-the-counter drugs taken regularly, prescription drugs currently prescribed and prescription drugs prescribed or refilled at the visit were recorded in addition to types of drugs, compliance issues, diagnoses rendered and sociodemographic information. Individual student data were aggregated and analyzed using Epi Info (Epidemiology Program Office of the Centers for Disease Control) and SPSS-PC. Results identified that: 1) the majority of patient visits involved the prescription of 1 2 drugs (88%); 2) major compliance issues included financial concerns, knowledge deficits, and complexity/demands of treatment; 3) commonly rendered diagnoses at drug visits for chronic conditions were hypertension and diabetes; for acute conditions, otitis sinusitis and upper respiratory infections; 4) anti-microbial agents, drugs used for relief of pain, and cardiovascular drugs account for 60% of drug mentions; and 5) the numbers of drugs prescribed or refilled at visits were similar by type of preceptor, except fewer single drugs were prescribed or refilled at visits supervised by nurse preceptors. Findings are discussed relative to deepening the understanding of advanced practice nursing education and the prescribing practices of NP students and their preceptors. PMID- 11033678 TI - Recognizing young people at risk for sudden cardiac death in preparticipation sports physicals. PMID- 11033679 TI - A study of the effectiveness of a primary care community clinic. PMID- 11033680 TI - Breast cancer genetics and the role of tamoxifen in prevention. PMID- 11033681 TI - Nurse practitioner, nurse midwife and physician assistant attitudes and care practices related to persons with HIV/AIDS. AB - Although multiple studies of nurses' attitudes toward people living with HIV/AIDS (PLWAs) can be found in the literature, little is known about the attitudes, beliefs and practices of nurse practitioners (NPs), certified nurse midwives (CNMs), and physician assistants (PAs). A survey including a 21-item AIDS Attitude Scale measuring the constructs of Avoidance and Empathy was sent to 1,291 NPs, CNMs and PAs in Louisiana, Arkansas and Mississippi to describe their attitudes and care practices related to PLWAs. Respondents who were more comfortable treating PLWAs had significantly lower avoidance scores and significantly higher empathy scores than respondents with lower comfort levels in providing care. Greater than 80% of respondents indicated that they would provide health care to HIV-infected individuals. Respondents who referred HIV/AIDS patients for all care did so primarily due to lack of experience with HIV and the availability of more experienced providers. Avoidance and empathy scores were not found to be significantly associated with referral for care. This study suggests that this group of providers has relatively low avoidance and high empathy toward PLWAs and is willing to care for HIV-infected individuals. PMID- 11033682 TI - Understanding nurse practitioner preparation. PMID- 11033683 TI - Care of the perioperative patient: a new dimension for advanced practice nurses. PMID- 11033684 TI - Cytolytic vaginosis: an overlooked cause of cyclic vaginal itching and burning. PMID- 11033685 TI - Improving self-care in adults with asthma using peak expiratory flow rate home monitoring. AB - Asthma management is based on step therapy incorporated into an individualized patient treatment plan. Medication selection is based on differing degrees of asthma severity. With proper assessment of the patient and a severity level incorporating the patient's needs, a clinician can create a credit card treatment plan for each patient. The assessment should include both PEFR and symptom monitoring as a means of incorporating the CDC's severity guidelines and treatment options into the credit card plan. Evaluation of technique, review of home monitoring outcomes, and reinforcement during clinic visits is likely to be helpful for those patients who do home monitoring. Note, however, that not all patients should be treated using this self-management approach. Asthma associated with comorbidities may be a reason to manage patients more closely either by clinic visit or telephone. Asthma in both older and pregnant patients presents issues of drug safety (Evans, Brown, & Morain, 1997). The common comorbidities of chronic obstructive pulmonary disease, sinusitis, GERD, cardiovascular disease and diabetes present unique issues of difficulty of diagnosis and drug safety. By following individualized asthma management plans, patients should be able to achieve prevention or reduction of chronic symptoms. They should also notice an improvement in physical activity, the reduction or elimination of exacerbations and improved overall satisfaction. PMID- 11033686 TI - Periodontal disease: diagnosis and management. AB - Approximately 50% of all adults in the United States have some form of gingivitis, and 80% have experienced some degree of periodontitis. Despite the use of fluoride, a critical element in the prevention of periodontal disease, many children and adults still suffer needlessly from this "silent" destroyer of the gum tissue and its underlying bone structures. This article highlights the prevalence, pathophysiology, classifications, health risk factors and clinical management of clients with periodontal diseases. Regardless of the health care setting, the advanced practice nurse plays a critical role in the prevention, early diagnosis and intervention of periodontal disease. PMID- 11033687 TI - The case for combined education for the NP and CNS. PMID- 11033688 TI - Cultural health care or child abuse? The Southeast Asian practice of cao gio. AB - Although the immigration of Southeast Asians more than doubled over the past 20 years, much misunderstanding still persists regarding traditional, culture-bound health practices used by Vietnamese, Cambodian and Laotian families. The research presented in this paper explains one such traditional practice, that of cao gio. Cao gio, a dermabrasion therapy, is used extensively by many Southeast Asian women to treat a variety of illnesses. Health care providers do not appreciate this practice, however, to the extent that it is claimed to be a form of child abuse in some of the literature reviewed. A better understanding of the practice by those involved in the primary care of Southeast Asian families is not only necessary, but essential to avoid the mislabeling and misinterpretation of cultural, traditional health practices as child abuse. Lack of awareness of Southeast Asian cultural practices is quite significant, even though the United States experienced three major waves of Southeast Asian immigration during the past 20 years (Takeuchi, & Young, 1994). Over the past decade, the number of Southeast Asian immigrants and refugees to the United States has increased 108% or by more than 1 million individuals (Fong, & Mokuau, 1994; Frank-Stromborg, 1991; Ja, & Aoki, 1993; Tran, 1991; U.S. Census Bureau, 1998). Yet, despite this, Southeast Asian individuals are poorly understood, and the literature reveals little about what health, illness and treatment constitute for these different cultures (Boston, 1992; Buchwald, Panwala, & Hooton, 1992). The ethnic groups that comprise Southeast Asians, formerly known as the Indochinese, include individuals from Vietnam, Cambodia and Laos (Catanzaro, & Moser, 1982; Chung, & Kagawa-Singer, 1993). The majority of these individuals are refugees who fled their countries due to political persecution (Toole, & Waldman, 1993). This paper describes the findings of a phenomenological research study completed among 19 Southeast Asian women regarding beliefs about health and health practices. The women discussed the practice of cao gio (phonetically pronounced as gow yaw) and its importance, effectiveness and significance to their cultures. The discussion relates the study findings to the debate and issues surrounding child abuse, the consequences of misreporting, as well as the consequences of cultural ignorance in the provision of health care to those of different cultural practices and beliefs, particularly those of Southeast Asian ethnicity. PMID- 11033689 TI - Elder care: a positive approach to caregiving. AB - The primary focus of caregiving research over the past two decades has centered on the negative aspects of the caregiving process. The purpose of this paper is to show that there are positive aspects to caregiving and the importance of refocusing attention to this area. Lack of information regarding the positive dimensions of caregiving alters perceptions of the caregiving experience and limits the ability to enhance theory related to caregiver adaptation. Understanding positive aspects of caregiving may help clinicians and practitioners work more effectively with the family caregiver. PMID- 11033690 TI - Polycystic ovary syndrome. AB - Polycystic ovary syndrome is a complex endocrine disorder of hypothalamic pituitary dysfunction that presents with anovulation, hirsutism and infertility. Women with PCOS have increased risk for developing NIDDM, dyslipidemia and premature cardiovascular disease. Because of its vague presentation and potential for numerous complications, PCOS should be evaluated carefully. Unfortunately, there is no cure for PCOS, and the treatment has numerous limitations. Weight loss in an obese patient may greatly improve the patient's response to treatment and should be encouraged. PMID- 11033691 TI - Medical errors: medical-surgical nurses speak. PMID- 11033692 TI - Syndrome of inappropriate antidiuretic hormone secretion: recognition and management. AB - Older persons are at greater risk for developing hyponatremia. Renal system changes which occur with aging cause alterations in the functional balance of fluid and electrolytes, and sodium regulation is not as effective. In one form of hyponatremia seen among the elderly, total body sodium content is normal, but water is retained because of increased antidiuretic hormone (ADH) secretion. This condition, called the syndrome of inappropriate antidiuretic hormone secretion (SIADH), has many causes including chronic illnesses and various medications. The physiology behind SIADH, its causes, diagnostics, and various treatment modalities are explored. PMID- 11033693 TI - The use of telemetric oximetry in the clinical setting. AB - The reliability and ease of use of a combined functions-cardiac telemetry and telemetric pulse oximetry system was evaluated in a four phase investigation. This technology was assessed favorably for use in the care of patients on continuous epidural and patient-controlled analgesia. PMID- 11033694 TI - Promoting outcomes for patients with spinal cord impairments and ostomies. AB - Patients with spinal cord impairments (SCI) and ostomies have unique needs which must be addressed by medical-surgical nurses. By considering the anatomy and physiology of SCI, functional ability, cognitive limitations, and a myriad of other factors, nurses can positively influence clinical and social outcomes. PMID- 11033695 TI - An alternative to sutures. AB - Cyanoacrylate tissue adhesives are used in numerous settings to close wounds without the use of sutures. The product and its use for closing chest tube wounds is discussed. PMID- 11033696 TI - Pharmacologic prevention and treatment of osteoporosis in women. PMID- 11033697 TI - Reimbursement issues in advanced practice nursing: an overview. AB - In this era of rapid change and restructuring of the health care delivery system, APNs are faced with many new opportunities and challenges. This implies that APNs should know the federal and state laws, rules, and regulations that clearly define their scope of practice and prescriptive authority. They should familiarize themselves with Medicare and Medicaid payment policies, and the state laws related to the private insurance practices. By learning the basic language of third-party payers, APNs will find themselves in a better position of understanding the process that ensures reimbursement for the services rendered. Thus, APNs should keep up with the new demand for their expertise by having close ties with new knowledge and new technologies. In addition, APNs should have the skills, credentials, and the knowledge of computer technology for communication, distance learning, diagnosis, and treatment. The ultimate goal is to provide high quality and cost-effective care. PMID- 11033698 TI - Retention of nurses: an organizational priority. PMID- 11033699 TI - Nursing's hazards to health. PMID- 11033700 TI - The efficacy of carotid endarterectomy: a vascular surgery perspective reducing hospital stay. AB - Contemporary vascular management results in significant reduction of stroke risk. Carotid endarterectomy is both safe and effective, often with a 1 or 2-day hospital stay. State-of-the-art nursing practice is key to the efficacy of patient management. PMID- 11033701 TI - Measurement of gastric residual volume: state of the science. AB - This literature review substantiates that there is confusion regarding what constitutes excessive gastric residual volume in adult tube-fed patients. Only one research study regarding gastric residuals was located; it indicated that 200 ml for a nasogastric tube and 100 ml for a gastrostomy tube should cause concern. The implications of these findings for adult-health nurses are discussed. PMID- 11033702 TI - Surviving managed care: the effect on job satisfaction in hospital-based nursing. AB - Major changes brought about by managed care have redefined the nursing profession. Current trends such as case management, downsizing, restructuring of the workforce, and changes in the patient profile have had numerous effects, particularly on job satisfaction among hospital-based nurses. Strategies to improve job satisfaction during this era of increased managed care penetration include enhanced communication mechanisms, support from hospital administration, implementation of care models that promote professional nursing practice, adequate staffing, and competitive salaries and benefits. PMID- 11033703 TI - An analysis of the epidemiology of latex allergy: implications for primary prevention. AB - The prevalence of latex allergy is escalating due to increased latex glove use following the institution of universal precautions. The purpose of this article is to review research pertaining to latex allergy, summarize national guidelines relating to latex allergy, and discuss the implications for nursing practice, education, and public policy. PMID- 11033704 TI - Suffering of critical care nurses with end-of-life decisions. AB - End-of-life situations occur in critical care units daily, exposing nurses to these situations on a regular basis. These critical care nurses experience emotions similar to the emotions identified as suffering. There is limited information regarding how nurses deal with these emotions or the long-term consequences of continued, unrelieved suffering. This study has implications for all nurses who care for seriously and critically ill adults. PMID- 11033705 TI - Vaccination: refuting the refusals. PMID- 11033706 TI - The effects of stress on oncology staff. PMID- 11033707 TI - The validity of a sore throat score in family practice. AB - BACKGROUND: Reducing the number of antibiotic prescriptions given for common respiratory infections has been recommended as a way to limit bacterial resistance. This study assessed the validity of a previously published clinical score for the management of infections of the upper respiratory tract accompanied by sore throat. The study also examined the potential impact of this clinical score on the prescribing of antibiotics in community-based family practice. METHODS: A total of 97 family physicians in 49 Ontario communities assessed 621 children and adults with a new infection of the upper respiratory tract accompanied by sore throat and recorded their prescribing decisions. A throat swab was obtained for culture. The sensitivity and specificity of the score approach in this population were compared with previously published results for patients seen at an academic family medicine centre. In addition, physicians' prescribing practices and their recommendations for obtaining throat swabs were compared with score-based recommendations. RESULTS: Of the 621 cases of new upper respiratory tract infection and sore throat, information about prescriptions given was available for only 619; physicians prescribed antibiotics in 173 (27.9%) of these cases. Of the 173 prescriptions, 109 (63.0%) were given to patients with culture-negative results for group A Streptococcus. Using the score to determine management would have reduced prescriptions to culture-negative patients by 63.7% and overall antibiotic prescriptions by 52.3% (both p < 0.01). Culturing of throat samples would have been reduced by 35.8% (p < 0.01). There was no statistically significant difference in the sensitivity or specificity of the score approach between this community-based population (sensitivity 85.0%, specificity 92.1%) and an academic family medicine centre (sensitivity 83.1%, specificity 94.3%). INTERPRETATION: An explicit clinical score approach to the management of patients presenting with an upper respiratory tract infection and sore throat is valid in community-based family practice and could substantially reduce the unnecessary prescribing of antibiotics for these conditions. PMID- 11033708 TI - Investigation and treatment of osteoporosis in patients with fragility fractures. AB - BACKGROUND: Many patients who have undiagnosed osteoporosis and a recent fragility fracture present to fracture clinics in Canadian hospitals, where the focus of management is on fracture care. The rate of diagnosis and treatment of osteoporosis in this patient group is unknown. METHODS: Patients who presented with fractures at sites consistent with fragility-type fractures were identified through a retrospective chart review of fracture clinic visits in 3 Ontario community hospitals in selected weeks in February and November 1996 and August and May 1997. These patients were contacted by mail and telephone follow-up to obtain consent to participate in a telephone interview. Patients were excluded if the index fracture had been traumatic, if they were younger than 18 years, or if they had medical conditions known to be associated with secondary bone loss. Eligible patients were questioned about their history of prior fractures, diagnosis of osteoporosis, and investigation and treatment of osteoporosis before or after the index fracture. RESULTS: Among 2694 fracture clinic visits, we identified 228 patients (8.4%) with fragility-type fractures. Of the 228, 128 (56.1%) were contacted and agreed to participate in an interview about 1 year from the date of the index fracture. Of the 128 patients, 108 (83 postmenopausal and 13 premenopausal women and 12 men) were confirmed as eligible. Of the 108, 43 had experienced 53 fractures in addition to the index fracture in the preceding 10 years, of which 71% were of the fragility type. At interview, only 20 (18.5%) (all postmenopausal women) of the 108 patients reported that they had received a diagnosis of osteoporosis. Of the 20, 90% and 45% respectively had been advised to take calcium and vitamin D supplements; 8 (40%) were receiving hormone replacement therapy (HRT), and 8 (40%) were taking bisphosphonates. Of the 88 patients who had not received a diagnosis of osteoporosis, 4 (4.5%) were receiving HRT, none was taking bisphosphonates, and less than 20% had been advised to take supplemental calcium or vitamin D. INTERPRETATION: In a representative sample of patients at urban fracture clinics, less than 20% who presented with a fragility-type fracture had undergone investigation and adequate treatment of osteoporosis at 1-year follow-up. Since previous fracture significantly increases the risk for future fracture, this clearly is a deficiency in management. Through improved identification and treatment of patients with osteoporosis-related fractures who present to fracture clinics, there is a significant opportunity to reduce the rates of illness and death associated with this condition. PMID- 11033710 TI - Prevention of perinatal transmission of HIV infection. PMID- 11033709 TI - Expanding the epidemiologic profile: risk factors for active tuberculosis in people immigrating to Ontario. AB - BACKGROUND: Many people immigrating to Canada come from countries with a high burden of tuberculosis. The aim of this study was to develop a detailed epidemiologic profile of foreign-born people with tuberculosis living in Ontario. METHODS: In this population-based case-control study, cases of tuberculosis diagnosed in 1994-1995 were identified from the database of the Ontario Reportable Disease Information Service and were considered eligible for analysis if a record of landing (receipt of permission to establish residence in Canada) from the period 1986-1995 was found in the Citizenship and Immigration Canada (CIC) database, if the person was at least 11 years of age at the time their visa was issued, and if the person had not been diagnosed with tuberculosis before becoming legally landed in Canada. Control subjects, who met the same criteria as the case subjects but who did not have tuberculosis in 1994-1995, were identified from a CIC database for landed immigrants. RESULTS: A total of 1341 cases of tuberculosis in foreign-born people were reported in Ontario in 1994-1995. A record of landing was found in CIC databases for 1099 of these people, 224 of whom were not legally landed at the time of diagnosis. In total, 602 cases met the inclusion criteria. The 2 strongest determinants of risk among those who had become landed within the preceding 10 years were referral for medical surveillance by immigration officials (odds ratio [OR] 3.8, 95% confidence interval [CI] 2.6-6.0) and world region of origin (Somalia [OR 67.7, 95% CI 31.3 154.9], Vietnam [OR 25.0, 95% CI 12.5-50.0], the Philippines [OR 11.9, 95% CI 6.0 23.3], other sub-Saharan African countries [OR 11.6, 95% CI 5.7-23.2], India [OR 9.7, 95% CI 4.9-18.9], China [OR 6.1, 95% CI 3.1-12.1], other Asian countries [OR 4.7, 95% CI 2.4-9.1], the Middle East [OR 4.1, 95% CI 2.0-8.3], Latin America [OR 1.9, 95% CI 0.9-3.8), and the former socialist countries of Europe [OR 1.8, 95% CI 0.8-3.8]; the reference category was countries with established market economies). Low socioeconomic status was an independent risk factor. INTERPRETATION: The risk of tuberculosis in groups of people migrating to Ontario is highly variable and is influenced by several factors. Successful population based tuberculosis prevention strategies will need to accommodate this variability. PMID- 11033711 TI - Bioethics for clinicians: continuing the series. PMID- 11033712 TI - Why Sackett's analysis of randomized controlled trials fails, but needn't. PMID- 11033713 TI - Equipoise, a term whose time (if it ever came) has surely gone. PMID- 11033714 TI - Evidence-based medicine: a commentary on common criticisms. AB - Discussions about evidence-based medicine engender both negative and positive reactions from clinicians and academics. Ways to achieve evidence-based practice are reviewed here and the most common criticisms described. The latter can be classified as "limitations universal to the practice of medicine," "limitations unique to evidence-based medicine" and "misperceptions of evidence-based medicine." Potential solutions to the true limitations of evidence-based medicine are discussed and areas for future work highlighted. PMID- 11033715 TI - Bioethics for clinicians: 18. Aboriginal cultures. AB - Although philosophies and practices analogous to bioethics exist in Aboriginal cultures, the terms and categorical distinctions of "ethics" and "bioethics" do not generally exist. In this article we address ethical values appropriate to Aboriginal patients, rather than a preconceived "Aboriginal bioethic." Aboriginal beliefs are rooted in the context of oral history and culture. For Aboriginal people, decision-making is best understood as a process and not as the correct interpretation of a unified code. Aboriginal cultures differ from religious and cultural groups that draw on Scripture and textual foundations for their ethical beliefs and practices. Aboriginal ethical values generally emphasize holism, pluralism, autonomy, community- or family-based decision-making, and the maintenance of quality of life rather than the exclusive pursuit of a cure. Most Aboriginal belief systems also emphasize achieving balance and wellness within the domains of human life (mental, physical, emotional and spiritual). Although these bioethical tenets are important to understand and apply, examining specific applications in detail is not as useful as developing a more generalized understanding of how to approach ethical decision-making with Aboriginal people. Aboriginal ethical decisions are often situational and highly dependent on the values of the individual within the context of his or her family and community. PMID- 11033716 TI - Environment and health: 3. Ozone depletion and ultraviolet radiation. PMID- 11033717 TI - Setting priorities for waiting lists: defining our terms. Steering Committee of the Western Canada Waiting List Project. AB - In order to develop standardized measures to assess patients' relative priority for services for which there are waiting lists, it is essential that key terms be clearly defined. We propose that severity be defined as the degree or extent of suffering, limits to activities or risk of death; that urgency be defined as severity combined with considerations of the expected benefit and the natural history of the condition; that need be considered equivalent to urgency; and that priority be defined in terms of urgency (or need) with or without consideration of social factors. PMID- 11033718 TI - Patenting life: genetically altered mice an invention, court declares. PMID- 11033719 TI - MDs worried about access to care, CMA survey indicates. PMID- 11033720 TI - An interactive approach to treating prostate cancer. PMID- 11033721 TI - Desperate government begs Yellowknife's MDs to stay. PMID- 11033722 TI - West Nile fever heads north. PMID- 11033723 TI - New influenza drugs zanamivir (Relenza) and oseltamivir (Tamiflu): unexpected serious reactions. PMID- 11033724 TI - Intravenous Rh(o) [D] immune globulin [human] (WinRho SDF): suspected hemolytic/renal adverse reactions. PMID- 11033725 TI - Should nasal-paranasal-oronasopharyngeal lymphomas in children be classified differently from the other localizations? PMID- 11033726 TI - Thrombocytopenia with absent radii (TAR syndrome): new advances in the mechanism of thrombocytopenia. PMID- 11033727 TI - Department of pediatric hematology and oncology, University of Catania, Italy. PMID- 11033728 TI - Late cardiac damage of anthracycline therapy for acute lymphoblastic leukemia in childhood. AB - Long-term leukemia survivors (46) underwent cardiac evaluation, including physical examination, ECG, exercise testing, and echocardiography. They were 2-17 years old at diagnosis and 5-23 years old after treatment. Thirty-four survivors received anthracyclines (AC) (mean 203 mg/m2), 12 of them had also alkylating agents (AA) and 12 had no AC. Exercise tolerance was bellow predicted values in 21 (48%) survivors and 21 survivors had ECG abnormalities, which were more frequent in those treated with AC. Concomitant AC with AA was correlated with prolonged isovolumic relaxation time (IVRT) and influenced significantly the volume of left atrium (p = .02). Sixteen (52%) survivors had IVRT > or = 90 ms. There were no significant differences in other parameters of diastolic or systolic function. Despite the lack of clinical symptoms in the survivors treated with lower doses of AC, subtile abnormalities in myocardial function were found, mainly manifest as abnormal diastolic function. Prolonged IVRT may be a sensitive indicator for early detection of AC cardiotoxicity. PMID- 11033729 TI - Prevention of childhood leukemia by BCG vaccination in newborns? A population based case-control study in Lower Saxony, Germany. AB - Data from a case-control study in Lower Saxony, Germany, were used to assess whether the risk for childhood cancer may be reduced by bacille Calmette-Guerin (BCG) vaccination in the neonatal period. There were 420 newly diagnosed childhood cancer cases from the German cancer registry and 613 controls eligible for this study. A mailed questionnaire was completed during a telephone interview with parents. Details on the perinatal history were abstracted from the birth charts by nurses blinded to the children's case-control status. Complete information was available for 259 cases and for 323 controls. A total of 85% of the controls had been BCG vaccinated in the newborn period. The adjusted odds ratios for BCG vaccination were 0.90 (95% confidence interval; 0.51-1.61) for leukemia and 0.61 (95% confidence interval; 0.25-1.50) for other cancers. Based on these data the probability of a 50% or more reduction of more reduction of the cancer risk by BCG vaccination in the newborn period is small. The statistical power of this study, however, was not high enough to rule out a smaller, still relevant reduction in cancer risk. PMID- 11033730 TI - Immune thrombocytopenic purpura in childhood in Norway: a prospective, population based registration. AB - A prospective, population-based registration of children with immune thrombocytopenic purpura (ITP) was performed in Norway in 1996 and 1997. Ninety two cases were identified, indicating an incidence of 5.3 per 100,000 children under 15 years. The sex ratio (female/male) was 1.2/1. Fifty-six percent presented with cutaneous signs only. The lowest platelet count was < 20 x 10(9)/L in 91%. In spite of mild bleeding symptoms, medical treatment was given in 68%, in most cases (57/63) with intravenous immunoglobulin. A total of 41/44 patients with platelet counts of < or = 5 x 10(9)/L were treated, regardless of whether they had mucous bleedings or not. Eighteen percent had platelet counts < 150 x 10(9)/L at 6 months, and 9% at 12 months following diagnosis. One patient with therapy-resistant chronic ITP died 16 months after diagnosis from an anesthesia complication related to profound epistaxis. This study shows a relatively high incidence. As in other studies, there was a tendency to treat platelet counts rather than bleeding symptoms. PMID- 11033731 TI - Short-term follow-up of the nutritional status of children undergoing autologous peripheral blood stem cell transplantation. AB - A prospective longitudinal study was conducted to analyze the evolution of the nutritional status of 34 children (12 girls and 22 boys), aged 1.5-15.8 years (median age 9.06), undergoing autologous peripheral blood stem cell transplantation (PBSCT). The nutritional status was evaluated at baseline, days +1 and +7, discharge, and day +30 by dietary or parenteral intake, anthropometric and laboratory measurements, and nitrogen balance. At baseline, changes in anthropometric (53%) and biochemical measurements (83%) are frequent but mild. The mean caloric intake was normal. Children with normal values for the anthropometric parameters all had an intake > 80% (p < .01). No correlation was found between the anthropometric and biochemical parameters. During transplantation, significant changes (p < .001) were found for energy intake, albumin, transferrin, and nitrogen balance. Fibronectin, prealbumin, and retinol binding protein showed only a few changes. All but prealbumin recovered on day +30. No correlation was found between the nutritional status and toxicity or infection in children undergoing autologus PBSCT. The changes in the nutritional status observed at the start of transplantation correlated with the nutrional intake. Anthropometric and biochemical changes are complementary. The results may be ascribable to the fact that the patients in this series had mild malnutrition. PMID- 11033732 TI - Association between the change in nutritional status in response to tube feeding and the occurrence of infections in children with a solid tumor. AB - In 32 children with a solid tumor, the association between the change in weight for height, in response to 4 weeks of tube feeding during the intensive phase of treatment, and the occurrence of leukopenia, leukopenic infections, and nonleukopenic infections in a period thereafter (4-10 weeks) was studied. Factors possibly influencing the change in weight for height during the first 4 weeks of tube feeding were also assessed. A statistically significant negative correlation (rho = -0.59; p < .001) was found between the change in z-score of weight for height in response to the first 4 weeks of tube feeding, and the occurrence of nonleukopenic infections between 4 and 10 weeks. A reduced occurrence of nonleukopenic infections resulted in a significant reduction of the number of days of infection-related hospital admission (rho = .45; p = .009), which, besides providing advantages for the patient, also had economical benefits. The change in weight for height in response to tube feeding was mainly influenced by the incidence of therapy-induced vomiting (r = -.45; p = .02) and by the amount of energy provided by tube feeding (r = .47; p = .007). Based on these findings, it is recommended that naso-gastric tube feeding be used in children with a solid tumor during the early intensive phase of treatment, and that one should aim for a considerable increase in weight for height during the first 4 weeks of administration, since this has been shown to reduce the number of nonleukopenic infections in a subsequent period. The increase in weight for height may be improved by providing an optimal antiemetic protocol, which will increase energy uptake, and an energy-enriched formula, which will increase energy intake. PMID- 11033733 TI - Is AML1/ETO gene expression a good prognostic factor in pediatric acute myeloblastic leukemia? AB - To assess the clinical significance of AML1/ETO gene detected by nested reverse transcriptase polymerase chain reaction, the outcome of 7 patients with acute myeloblastic leukemia between 3 and 14 years of age were presented. All patients had complete remission (CR) at the end of induction (AML-MRC 10 protocol) and 4 underwent unpurged autologous, 2 allogeneic (from matched siblings) non-T-cell depleted bone marrow transplantations (BMT) in first CR. One patient died due to allogeneic BMT-related complications, and 4 patients relapsed at 13, 17, 18, and 26 months. Only one patient achieved second CR. All relapsed patients died between 18 and 36 months with resistant disease (n = 3) or infection during salvage chemotherapy (n = 1). Two patients who had autologous BMT are alive and disease free at 44 and 50 months. Although statistical significance could not be shown, event-free survival and overall survival rates of AML1/ETO-positive patients (28.57 and 28.57%, respectively) at 3.5 years were even lower than those of AML1/ETO-negative patients. The results confirm some previous reports that AML1/ETO gene in children and adolescents is not a favorable prognostic factor. PMID- 11033734 TI - Incidence of seizures in pediatric cancer patients treated with imipenem/cilastatin. AB - Proconvulsive tendency of imipenem/cilastatin is one of its well-known side effects. Most studies report incidence rates ranging from 1.5 to 3%. There is no study on the incidence rate among children with systemic malignancies. Eighty-two patients with various malignancies who received imipenem/cilastatin 143 times for neutropenic fever between March 1994 and October 1999 in Department of Pediatric Oncology, Gazi University, were identified. Three of these patients had convulsions attributed to imipenem/cilastatin; 3.6% of the patients had seizure, or 2% of imipenem/cilastatin administrations was followed by a seizure attack. PMID- 11033735 TI - Papillary adenocarcinoma of the thyroid in a thyroglossal duct cyst. AB - A case report is presented of an 8-year-old boy who underwent resection of a thyroglossal duct cyst to illustrate a rare, but significant, complication of a common clinical problem. Pathological examination revealed that it contained a papillary adenocarcinoma of the thyroid, presumably arising from ectopic glandular tissue in the cyst. Thyroglossal duct cysts are a common cause of midline neck masses in children. Occult thyroid carcinoma is a rare co-morbid finding. It infrequently leads to death, but thyroglossal duct cysts may also contain the only functioning, albeit ectopic, thyroid tissue. Patients with clinical thyroglossal duct cysts should be carefully evaluated preoperatively for the presence of tumor and other functioning thyroid tissue prior to excision of the cyst. PMID- 11033736 TI - Nasopharyngeal carcinoma in a boy with fragile X syndrome. AB - The authors report on what they believe to be the first case of nasopharyngeal carcinoma in a patient with fragile X syndrome. In the literature, a few cases of fragile X syndrome associated with malignancies are described. Although this association seems more than coincidental, the molecular correlation between this syndrome and cancer is yet unclear. PMID- 11033737 TI - Application of nitric oxide for a case of veno-occlusive disease after peripheral blood stem cell transplantation. AB - A 5-year-old girl at high risk for acute lymphoblastic leukemia was treated with high-dose chemotherapy and autologous peripheral blood stem cell transplantation (PBSCT). However, her condition was complicated by veno-occlusive disease of the liver (VOD) after PBSCT. For treatment of VOD, transdermal isosorbide tape was applied as a nitric oxide (NO) donor. The signs of VOD improved immediately after NO treatment was initiated, and the patient showed no side effects from the transdermal isosorbide tape. PMID- 11033738 TI - Effects of food supplementation and education on folate stores. PMID- 11033739 TI - Does your clinical investigator understand the consequences of non-compliance? AB - The increasing number and complexity of clinical trials, coupled with the recent increase in attention and enforcement from the U.S. Food and Drug Administration, arguably makes it more important than ever before that clinical investigators understand the regulatory requirements with which they must comply. This article outlines some of these requirements, common areas of non-compliance and FDA's enforcement options in dealing with such non-compliance. Finally, the article provides some suggestions for attaining and managing investigator compliance. PMID- 11033740 TI - A comparison of the U.S. EPA FIFRA GLP standards with the U.S. FDA GLP standards for nonclinical laboratory studies. AB - In the early 1980's both the Environmental Protection Agency and the Food and Drug Administration were receiving studies on the products that they regulate that were unscientifically sound, some even being fraudulent. Studies were being submitted that had not been done under sound scientific practice; data were missing; necessary documentation to reconstruct the study, also known as an audit trail, was not in place, it was this evidence that lead to the Good Laboratory Practice Standards (GLPs) being codified and becoming regulation. The GLPs were meant to assure that studies submitted to the agencies for the registration of products for which they were responsible would protect the environment, and, the safety and health of the public. PMID- 11033741 TI - A comparison of the U.S. FIFRA GLP standards with the Japan MAFF GLP standards. AB - Side-by-side comparisons are an effective way to compare different GLP standards. These comparisons allow easy elucidation of differences in the regulations. A side-by-side comparison of the EPA, GLPs and the IMAFF GLPs was constructed and evaluated. The major differences are reported. PMID- 11033744 TI - [Pediatric Sleep Medicine 2000. Vienna, 24-26 February 2000. Abstracts]. PMID- 11033742 TI - Confronting actual practice with practice guidelines in oncology. AB - Improving health care quality requires the availability of data to identify and eliminate unnecessary variations in the care process. Variations can be caused by an ineffective implementation of research findings or by obstacles to the translation of research into clinical practice. The analysis of current patterns of care by the use of routine data from electronic patient records or clinical registries may help highlight these deficiencies in actual care. The growing infrastructure of information technologies and the knowledge about clinically relevant variations of routine practice may help us understand the mechanisms that are impeding the translation of research into practice. There is a need to scrutinize these variations of practice and the barriers to guideline implementation. We think that an understanding and open discussion of such reasons may help, to continuously improve the quality of patient care. This process facilitates efforts and strategies to implement evidence-based medicine in the daily routine. PMID- 11033743 TI - Findings and actions by the IR-4 Project in response to a bushel of EPA inspections. AB - The IR-4 Project is dedicated to all aspects of minor crop pest management, in cooperation between USDA, universities, and other state or Federal entities. Human safety decisions made by EPA are based upon field raw data, analytical reports, and scientific review and interpretation. Quality Assurance personnel, an independent body of internal inspectors and auditors, assist both regulatory inspector and on-site field personnel during EPA site inspections. This article contains information gleaned from a large number of university and other test site inspections, which were conducted by EPA to fulfill requirements of Good Laboratory Practice regulations, 40 CFR 160. PMID- 11033745 TI - [20th Yugoslav Surgical Congress. Zlatibor, 17-21 May 1998. Abstracts]. PMID- 11033746 TI - Promoting mental health after childbirth: a controlled trial of primary prevention of postnatal depression. AB - OBJECTIVES: To investigate the effectiveness of a psychosocial intervention for the prevention of postnatal depression. DESIGN: A controlled trial. METHOD: Women expecting their first or second child and designated as 'more vulnerable' by the Leverton Questionnaire (LQ) or Crown Crisp Experiential Index (CCEI) were allocated to a preventive intervention (N = 47) or control group (N = 52) by expected date of delivery to provide groups expecting their babies around the same time. Women were assessed at 3 months postnatal. An additional group of women designated as 'less vulnerable' (N = 88) were assessed to confirm the validity of the LQ as a vulnerability measure. RESULTS: Questionnaire measures of mood in first-time mothers invited to the Preparation for Parenthood groups revealed significantly more positive mood than in the group receiving routine care. The median Edinburgh Postnatal Depression Scale (EPDS) score for those invited was 3, compared to 8 for those not invited (p < .005). The diagnosis of depression using the Present State Examination revealed differences for both groups, though it reached statistical significance only with the first-time mothers. Only 19% of the 'more vulnerable' invited first-time mothers were 'borderline' or 'cases' at any time in the first 3 months postnatally compared to 39% of those not invited. The Surviving Parenthood groups for second-time mothers were not successful. CONCLUSION: Some depressions following childbirth can be prevented by brief interventions that can be incorporated with existing systems of antenatal classes and postnatal support groups. PMID- 11033747 TI - Attitudes to emotional expression and personality in predicting post-traumatic stress disorder. AB - OBJECTIVES: To test hypotheses derived from a suggestion of Williams (1989) that negative attitudes towards emotional expression act as a predisposing or maintaining factor for post-traumatic stress reactions following a traumatic event. DESIGN: The study employed a prospective design in which attitudes to emotional expression, the 'Big Five' personality factors (Costa & McCrae, 1992a) and initial symptoms and injury severity within 1 week of a road traffic accident were used to predict the development of post-traumatic stress disorder 6 weeks post-accident. METHOD: Sixty victims of road traffic accidents randomly selected from attenders at a large A&E department were assessed by questionnaire and interview. Measures comprised a 4-item scale relating to emotional expression, standardized scales for intrusion and avoidance features of traumatic experiences, and for anxiety and depression and the NEO-FFI Five Factor Personality Inventory. Forty-five of these participants responded to a postal questionnaire follow-up. In this survey the battery was repeated and also included a self-report diagnostic measure of post-traumatic stress disorder (PTSD). RESULTS: The percentage of the sample meeting DSM-IV diagnostic criteria for PTSD at 6 weeks post-trauma was 30.8%. A small but significant relationship was found for negative attitudes to emotional expression at 1 week to predict intrusive symptoms and diagnosis at 6 weeks, over and above the independent relationships of initial symptoms, initial injury severity, personality and coping. The emotional expression measure was largely stable between the two points of measurement. More negative attitudes to emotional expression were related to less openness, extraversion and agreeableness personality domains. CONCLUSIONS: Some support for the hypotheses was found in relation to the development of PTSD and for the status of attitudes to emotion as a stable trait related to personality factors. The potential importance of attitudes to emotional expression in therapy and other work is discussed. PMID- 11033748 TI - Self-discrepancies and causal attributions: studies of hypothesized relationships. AB - The self-concept and causal attributions are both centrally implicated in psychological disorders including depression and paranoia. In two investigations of the dynamic relationships between causal attributions and self representations, non-patient participants completed questionnaires derived from Higgins' (1987) Self-Discrepancy Theory before and after completing a measure of causal attribution. In Study 1, consistent with cognitive models of depression, external attributions for negative events were associated with reductions in self actual:self-ideal discrepancies. Study 2 revealed significantly different effects on self-discrepancies of three types of causal attributions. Internal attributions led to increased self-actual:self-ideal discrepancies as well as increased discrepancies between self-perceptions and the believed views of others about the self (self-actual:other-actual discrepancies). External situational attributions led to no changes in either self-actual:self-ideal or self actual:other-actual discrepancies. External personal attributions led to no changes in self-actual:self-ideal discrepancies but increased self-actual:other actual discrepancies. These findings point to the value of distinguishing between different kinds of external attributions. They show that self-representations and causal attributions are closely coupled cognitive domains. The results also suggest that paranoid ideation might be specifically associated with external personal attributions for negative events. PMID- 11033749 TI - Pre-sleep cognitive activity: a comparison of sleep-onset insomniacs and good sleepers. AB - OBJECTIVE: Pre-sleep cognitive activity has been implicated in the maintenance of sleep-onset insomnia. The present study aimed to investigate the focus of attention, content and characteristics of cognition during the pre-sleep period. METHOD: A semi-structured clinician-administered interview designed to index the content of pre-sleep cognition was completed by individuals diagnosed with sleep onset insomnia (N = 30) and good sleepers (N = 30). RESULT: The pre-sleep cognitive activity of insomniacs could be distinguished from that of good sleepers by being more focused on worries, problems and noises in the environment, and less focused on 'nothing in particular'. In terms of content, the insomnia group were more likely to think about not sleeping or about something that had happened during the day. Insomniacs experienced their pre sleep cognitive activity as more occupying, less intentional, for a longer duration, and as causing more difficulty with sleep onset compared to good sleepers. Pre-sleep imagery was reported at similar rates across diagnosis, but was more distressing and more likely to be associated with strong physical sensations for the insomniac group compared with the good sleeper group. CONCLUSION: The present study provides a comprehensive investigation of pre-sleep cognitive activity and raises a number of areas for future research including monitoring of bodily sensations, imagery, problem-solving and non-active strategies in facilitating sleep onset. PMID- 11033750 TI - Compliance and psychological reactance in schizophrenia. AB - OBJECTIVES: Non-compliance with neuroleptic medication in schizophrenia is a major cause of relapse. A number of sociodemographic variables, and illness, attitudinal and treatment variables, have been demonstrated to be associated with non-compliance. The present study examined a range of these variables and their predictive value in determining past and current compliance. METHODS: Thirty-nine patients suffering from schizophrenia and three patients suffering from schizoaffective disorder completed a series of questionnaires assessing psychological reactance, insight, subjective response to medication, perceived threat to freedom of choice, and degree of current and past compliance. Logistic regression analyses were performed to determine which factors best predicted past and current compliance. RESULTS: Psychological reactance and age were found to be the best predictors of past compliance, with an interaction between reactance and perceived threat to freedom of choice posed by treatment provision also making a significant contribution. Past compliance behaviour and subjective response to medication predicted current compliance most significantly. CONCLUSIONS: Reactance is an important predictor of compliance history especially when patients perceive treatment to be a threat to freedom of choice. Subjective response to neuroleptics is most important in predicting current compliance. Implications for intervention are discussed. PMID- 11033751 TI - The National Adult Reading Test as a measure of premorbid IQ in schizophrenia. AB - OBJECTIVES: To investigate the validity of the NART as an estimate of premorbid IQ in schizophrenia. DESIGN: A within-in participants, follow-back design was adopted. METHODS: A sample of adults with schizophrenia who had presented to psychiatric services and had a measure of IQ routinely taken during childhood were traced and subject to follow-up WAIS-R and NART IQ assessment (N = 24). Measures of current IQ and NART estimated premorbid IQ were compared with the measure of IQ taken 'premorbidly', i.e. in childhood. RESULTS: There were no significant differences between childhood and adult measures of IQ. However there were significant differences between these two indices and NART estimated IQ, particularly where IQ deviated from general population means. The Vocabulary subtest of the WAIS-R performed better as an estimate of both premorbid and current IQ in the sample. CONCLUSION: Use of a word-reading test such as the NART to predict past levels of intellectual function should proceed with caution, particularly where IQ does not fall in the 'average' category. Use of more than one index of prior level of function is recommended. PMID- 11033752 TI - Sexual maturation and control issues among sexually abused and non-abused anorexia patients. AB - OBJECTIVES: To assess the relative salience of the maintenance of control and the avoidance of sexual maturation as sources of motivation for maintaining pathological eating behaviours among sexually abused anorexic patients. DESIGN/METHODS: A two-factor mixed experimental design was employed. Three independent groups (sexually abused anorexics (N = 12); non-abused anorexics (N = 9); non-anorexic/non-abused controls (N = 12)) constituted the between-subjects factor. Allocation to abuse/non-abuse group was dependent upon replies to a questionnaire-based measure of unwanted sexual experience. The within-subjects factor comprised three conditions in which words of various colours were presented to participants for colour-naming (Stroop, 1935). The conditions were represented by lists of neutral words, sexual maturation words, and control related words. Two trials were carried out in each condition and mean response times were measured. RESULTS: Within-group analyses revealed that interference was greater from sexual maturation words than from control-related words among the sexually-abused anorexics, but of equal magnitude among non-abused counterparts. Between-groups analyses found that abused patients experienced marginally greater cognitive interference from sexual maturation words than the non-abused patients. CONCLUSION: Theoretically, support is offered for elaborated schematic models of cognitive processing. Clinically, treatment interventions may need to pay particular attention to issues of sexual maturation among sexually abused anorexic patients. PMID- 11033753 TI - Screening for memory problems in multiple sclerosis. AB - OBJECTIVES: To assess the Brief, Repeatable Battery of Neuropsychological Tests (BRB-N) as a screening measure for memory problems in people with multiple sclerosis (MS). DESIGN: Sensitivity and specificity values were calculated and Receiver Operating Characteristic (ROC) curves plotted. METHODS: Patients were recruited from the regional MS Management Clinic. The BRB-N and WMS-R were administered. RESULTS: Using the Symbol Digit Modalities Test (SDMT) and 10/36 Spatial Recall Test (10/36 SRT) Total Immediate recall, the BRB-N had a sensitivity of 93% and specificity of 48%. CONCLUSION: The BRB-N is sensitive at detecting memory impairments, although its specificity is low. PMID- 11033754 TI - Sex-role traits and self-monitoring as dimensions of control: women with bulimia nervosa vs. controls. AB - OBJECTIVE: The study investigated sex-role traits and self-monitoring styles as two dimensions of self-control amongst women with bulimia nervosa and normal controls. DESIGN AND METHODS: The participants were 30 women with bulimia nervosa and 40 normal controls who completed questionnaires, which assessed bulimic symptoms, sex-role traits, and Synder's Self-Monitoring Scale. RESULTS: Women with bulimia nervosa were found to identify more strongly than controls with negative feminine traits and Other Directedness, a style of self-monitoring that focuses on pleasing others. CONCLUSIONS: These findings reinforce the notion that women with bulimia nervosa lack assertiveness skills and that the strategies they have developed in their interactions with others can be described as a Negative Yielding style of self-control. PMID- 11033755 TI - Natural history of primary hyperparathyroidism. AB - Primary hyperparathyroidism has evolved into a disorder that is largely asymptomatic. Nevertheless, there is ample evidence of target organ effects even in asymptomatic patients. Recent data suggest that the disease is stable in most asymptomatic patients. Little change is observed in biochemical parameters or bone mineral density over time. A subgroup of asymptomatic patients shows biochemical evidence of disease progression, although, in the author's series, no overt clinical complications developed. Surgical cure is associated with biochemical normalization and increased bone density. Several important questions remain: What are the neuropsychiatric and cardiovascular features of the disease? Are these features progressive over time, and do they regress with cure? What implications do any cardiovascular manifestations have on mortality? Is there an increase in fractures associated with mild asymptomatic disease? Is there an increase in fractures at more cortical sites, with a decrease in vertebral fractures in affected postmenopausal women? Although ongoing targeted research should answer some of these questions, a large multicenter trial is necessary to provide the data needed concerning the natural history of primary hyperparathyroidism. PMID- 11033756 TI - Primary hyperparathyroidism. When to observe and when to operate. AB - The decision to recommend parathyroid surgery in patients with symptomatic primary hyperparathyroidism is clear. The decision to recommend parathyroid surgery in patients with asymptomatic primary hyperparathyroidism is clearer than it was 10 years ago. In addition to the NIH guidelines, the author believes that patients who have vertebral osteopenia, patients who have vitamin D deficiency, and patients who are in their perimenopausal years should undergo surgery. Although surgery may be an acceptable option even for patients who do not meet these guidelines, many patients can be safely managed without surgery. For these patients, monitoring is essential because a small number will show progression of disease over time. PMID- 11033757 TI - Surgical approaches to primary hyperparathyroidism. AB - Primary hyperparathyroidism, once thought to be a rare disease entity, is now a common problem. It can be diagnosed with nearly 100% accuracy. Surgical therapy is the only definitive cure for this disease, and normocalcemia is achieved in 95% of patients at initial operation when performed by an experienced surgeon. Even when the operation is initially unsuccessful, most of the patients with persistent disease can subsequently be cured. Although some clinicians have proposed that asymptomatic patients can be medically managed, the cost of such treatment, problems with patient compliance with long-term follow-up, the increased risk of premature death associated with primary hyperparathyroidism, and the low morbidity of operation support a liberal policy for exploration in most patients. The authors believe that nonoperative therapy should be limited to older patients with multiple comorbid conditions and minimal hypercalcemia and clinical manifestations. PMID- 11033758 TI - Familial hypocalciuric hypercalcemia and other disorders with resistance to extracellular calcium. AB - Cloning of the CaR has increased understanding of the normal control of mineral ion homeostasis and has clarified the pathophysiology of PTH-dependent hypercalcemia. Cloning of the CaR has enabled identification of FHH and NSHPT as inherited conditions with generalized resistance to Ca2+o, which is caused in many cases by inactivating mutations in the CaR gene. In most kindreds with FHH, there is resetting of Ca2+o to a mildly elevated level that does not require an increase in the circulating level of PTH above the normal range to maintain it. FHH is not accompanied by the usual symptoms, signs, and complications of hypercalcemia. The kidney participates in the genesis of the hypercalcemia in FHH by avidly reabsorbing Ca2+; consequently, there is no increased risk of forming urinary calculi in most cases. Generally, there is no compelling rationale for attempting to lower the level of Ca2+o in these patients to a nominal normal level. In contrast, in primary hyperparathyroidism, the Ca2+o resistance is limited to the pathologic parathyroid glands, and the rest of the body suffers the consequences of high circulating levels of calcium, PTH, or both. In this condition, removal of the offending parathyroid glands is often the treatment of choice. Parathyroidectomy may also be appropriate in disorders with generalized resistance to Ca2+o owing to inactivating CaR mutations in the following special circumstances: in selected families with FHH in which there is unusually severe hypercalcemia, frankly elevated PTH levels, or atypical features such as hypercalciuria; in cases of NSHPT with severe hypercalcemia and hyperparathyroidism; and in the occasional mild case of homozygous FHH owing to CaR mutations that confer mild-to-moderate resistance to Ca2+o that escapes clinical detection in the neonatal period. As discussed elsewhere in this issue, selective calcimimetic CaR activators are being tested in clinical trials, which potentiate the activation of the CaR by Ca2+o, thereby resetting the elevated set point for Ca2+o-regulated PTH release in primary and secondary hyperparathyroidism toward normal. It is hoped that these agents may become an effective medical therapy for the acquired Ca2+o resistance in primary and secondary hyperparathyroidism and perhaps for that present in the unusual cases of FHH and NSHPT, resetting the "calciostat" downward and thereby reducing Ca2+o and PTH toward normal. PMID- 11033759 TI - Medical approaches to primary hyperparathyroidism. AB - Medical therapy is useful in cases of acute primary hyperparathyroidism, patients with recurrent disease, and parathyroid carcinoma. Among the therapeutic agents that have been employed, oral phosphate, bisphosphonates, and estrogens have been successful. The newly described calcimimetic agents directly block secretion of parathyroid hormone from the glands and offer an important new approach to medical therapy of primary hyperparathyroidism. PMID- 11033760 TI - Multiple endocrine neoplasia type 1. AB - Combined clinical and laboratory investigations of MEN-1 have resulted in an increased understanding of this disorder, which may be inherited as an autosomal dominant condition. Defining the features of each disease manifestation in MEN-1 has improved patient management and treatment and has facilitated a screening protocol. Application of the techniques of molecular biology has enabled the identification of the gene causing MEN-1 and the detection of mutations in patients. The protein encoded by the MEN1 gene has been shown to be involved in the regulation of JunD-mediated transcription, but much still remains to be elucidated. Recent advances permit the identification of mutant MEN1 gene carriers who are at a high risk for this disorder and who require regular and biochemical screening to detect the development of endocrine tumors. PMID- 11033761 TI - Pseudohypoparathyroidism. New insights into an old disease. AB - The GNAS1 gene (chromosome 20q13.3) encodes the alpha subunit of the stimulatory G protein (Gs alpha) and at least three additional, alternatively spliced transcripts, XL alpha s, NESP55, and the antisense transcript AS. Gs alpha transcripts seem to be derived exclusively, at least in the renal cortex, from the maternal allele. XL alpha s and AS are transcribed only from the paternal allele, and NESP55 is transcribed only from the maternal allele. Numerous GNAS1 mutations have been identified in PHP-Ia and pPHP. Patients with either disorder show skeletal and developmental defects now referred to as AHO. Owing to paternal imprinting, that is, inactivation of the paternal allele, which may be tissue- or cell-specific, resistance toward PTH and, often, other hormones is only observed in patients with PHP-Ia. Patients with PHP-Ib show PTH-resistant hypocalcemia and hyperphosphatemia but no AHO. The abnormal regulation of mineral ion homeostasis is paternally imprinted, such as in PHP-Ia/pPHP kindreds, Gs alpha activity/protein is normal in fibroblasts and blood cells, and no GNAS1 mutations have been identified. Recent linkage studies have mapped the genetic defect responsible for PHP-Ib to chromosome 20q13.3, making it likely that mutations in distinct regions of the GNAS1 gene are the cause of at least three different forms of PHP. PMID- 11033762 TI - Disorders of phosphate metabolism. AB - Correct identification of the disorders of hypophosphatemia and hyperphosphatemia is important for determining therapy. Further research will provide insights into normal phosphate homeostasis, a complex and fascinating process. PMID- 11033763 TI - Vitamin D deficiency and disorders of vitamin D metabolism. AB - The disorders of vitamin D metabolism are inherited metabolic abnormalities involving mutations of the vitamin D receptor or enzymes involved in the metabolism of vitamin D to its biologically active form 1,25-dihydroxyvitamin D. Although these mutations are rare, studies in affected patients and animal models have helped to identify critical actions of vitamin D and the mechanism by which it exerts its effects. Vitamin D deficiency, however, is an increasingly recognized problem among the elderly and in the general population. Screening for vitamin D deficiency only in those patients with known risk factors will result in a large proportion of unrecognized affected patients. PMID- 11033764 TI - The parathyroid hormone-related protein. AB - Many physiologic roles of PTHrP are emerging. The protein functions locally in diverse tissues, often regulating the entry of cells into a differentiation pathway or acting as an epithelial signal in epithelial-mesenchymal interactions. To carry out these functions, PTHrP uses the receptor it shares with PTH or one of several PTHrP receptors that have evolved to recognize selectively the PTH like region of PTHrP or other domains. Thus, PTHrP is a polyhormone. An exquisite selectivity barrier allows PTHrP to carry out its local tissue functions at the same time PTH uses their shared receptor to regulate systemic calcium homeostasis. This barrier is breached under pathologic circumstances, such as when malignant tumors secrete enough PTHrP into blood to cause PTH-like effects, including hypercalcemia. Powerful genetic models that have been developed in the past 7 years promise to give continuing insights into the physiology and pathophysiology of PTHrP. PMID- 11033765 TI - Regulation of the rat BB1 RNA during normal rat lung development. AB - BB1 was recently cloned from the WI-38 human fetal lung cell line. Human BB1 (hBB1) is expressed by multiple tissues, including lung. Because inhibition of BB1 translation using antisense oligodeoxynucleotides resulted in prevention of G1 traversal in cultured cells, we hypothesized that BB1 gene expression would be regulated during lung development with greater expression during periods of active lung growth. To gain insight into the expression of BB1 during lung development, a rat BB1 (rBB1) homologue was cloned and used in Northern hybridization analyses and in situ hybridization histochemistry (ISHH). Northern hybridization analyses of fetal and postnatal rat lung demonstrate that rBB1 RNA abundance is relatively low on fetal days E17 through E19, with a small peak of expression occurring on fetal day E20, then increases at birth with peak expression in adult lung. ISHH correlates with the Northern hybridization data and reveals rBB1 RNA expression throughout lung from E17 to E21 in both epithelium and mesenchyme. In postnatal lung, more intense expression of BB1 was observed than in fetal lung, localizing BB1 transcripts to proximal and distal airways and mesenchymal cells surrounding airways. Proliferating cell nuclear antigen (PCNA) was identified in lung sections adjacent to those used for ISHH and it was found that BB1 expression was present in PCNA-positive cells; however, BB1 expression was not limited to PCNA-positive cells in either the fetal or postnatal periods. This was most apparent in adult (60-day) rat lung where essentially no PCNA-positive cells were detected, but intense BB1 expression was detected in airway epithelium and surrounding mesenchyme. These studies demonstrate developmental regulation of BB1 during lung development. The findings are consistent with BB1 action in cell growth-related processes of fetal and early postnatal lung; however, the distribution of BB1 expression in relation to PCNA localization suggests that BB1 participates in cellular functions in addition to cell proliferation. PMID- 11033766 TI - Expression of functional beta 2-adrenergic receptors in a rat airway epithelial cell line (SPOC1) and cell density-dependent induction by glucocorticoids. AB - The beta 2-adrenergic receptor (beta 2AR) signal transduction system regulates many key functions of airway epithelium. In this study, we have pharmacologically characterized the beta 2AR and determined the impact of glucocorticoids on beta 2AR gene transcription in SPOC1 cells, a continuous cell line derived from the tracheal epithelium of rats. [125I]Cyanoiodopindolol assays demonstrated that binding to SPOC1 cell membranes was saturable (Bmax = 62.6 +/- 6 fmol/mg protein) and of high affinity (Kd = 6.3 +/- 0.8 pM). From competition experiments, the rank order of potency of agonists (isoproterenol > epinephrine >> norepinephrine) and the high affinity (Ki = 0.37 +/- 0.05 nM) of the beta 2-selective antagonist ICI 118,551 suggested the predominance of the beta 2AR subtype. Two isoforms of the alpha subunit of Gs (45 and 52 kDa) were identified by Western blot analysis. Isoproterenol-stimulated cyclic AMP levels increased in a dose-dependent manner, confirming that SPOC1 cell beta 2ARs are functionally coupled to adenylyl cyclase. The effect of glucocorticoids on beta 2AR expression was assessed in radioligand and transient transfection assays. Dexamethasone treatment of SPOC1 cells increased both beta 2AR protein and beta 2AR-luciferase fusion gene expression 1.6- to 3.1-fold, with the greatest increase demonstrated in cells cultured at low density compared to cells grown at high density. PMID- 11033767 TI - Does lung retention of inhaled particles depend on their geometric diameter? AB - Experiments with a bolus technique suggest that retained fractions in the airways are dependent on the geometric diameter of the particles. This view has been adopted by the International Commission on Radiological Protection (ICRP) in its new human respiratory tract model (HRTM). The aim of the present study was to test this view by the use of an inhalation technique, in which particles with an aerodynamic diameter of about 6 microns are inhaled extremely (0.05 l/s) and as a result, the particles are deposited mainly in small ciliated airways. Nine healthy subjects inhaled on one occasion monodisperse 111In-labelled polystyrene particles (geometric diameter 6.05 microns, aerodynamic diameter 6.2 microns) and on another occasion monodisperse 111In-labelled Teflon particles (geometric diameter 4.47 microns, aerodynamic diameter 6.5 microns). Both particles were inhaled at 0.045 L/s and radioactivity in the body was measured after 0, 24, 48, and 72 hours as well as after 1, 2, and, for some subjects, also 3 weeks. The retention in the lungs at 24 hours was slightly lower for the Teflon particles (47%) than for the polystyrene particles (51%). From earlier experimental data with different particle sizes as well as from predictions with theoretical lung models, this difference is reasonably explained by the somewhat larger aerodynamic diameter of the Teflon particles. Clearance of the 2 particle types between 1 day and 2 weeks was similar within each individual as well as in the whole group. The differences between the clearance of 4.5 microns and 6 microns geometric diameter particles observed in the present experiment are significantly different (P < .01) from the differences seen in earlier shallow bolus experiments as well as from the differences for such particles calculated with the HRTM, i.e., our experiment does not support the hypothesis that the fraction retained after 1 day is dependent on the geometric diameter in the size range studied. PMID- 11033768 TI - Mucosal IL-12 is more effective than systemic IL-12 in augmenting IFN-gamma expression and inhibiting allergic lung eosinophilia in murine lungs. AB - The relative efficacy of mucosal (intratracheal) and systemic (intraperitoneal) delivery of interleukin (IL)-12 was evaluated in a mouse model of allergic lung eosinophilia. Mucosal administration of IL-12 achieved 100- to 600-fold higher bronchoalveolar lavage (BAL) levels of IL-12, but 2- to 10-fold lower serum levels compared to systemic administration. Whereas both mucosal and systemic IL 12 inhibited BAL eosinophil recruitment at high doses (100-1000 ng), only mucosal IL-12 was effective at low doses (1-10 ng). Mucosal, but not systemic, administration of 1000 ng of IL-12 increased interferon (IFN)-gamma expression in BAL cells. In a model of ongoing eosinophilic inflammation, when mucosal or systemic IL-12 doses were initiated prior to peak eosinophilia, further eosinophil recruitment was inhibited. However, when IL-12 treatment was initiated after peak eosinophil recruitment occurred, recovery from eosinophilic inflammation was not facilitated. Our findings are the first to demonstrate that locally administered IL-12 inhibits eosinophil recruitment at 100-fold lower doses than systemic IL-12. The most likely mechanism of this enhanced inhibitory activity is a sustained increase in lung levels of IL-12 that augments IFN-gamma production from BAL cells. We suggest that future studies should evaluate the efficacy of low doses of nebulized IL-12 in inhibiting eosinophilic lung inflammation in asthma. PMID- 11033769 TI - Mucosal IL-12 inhibits airway reactivity to methacholine and respiratory failure in murine asthma. AB - The worldwide incidence, prevalence, and fatality rates from asthma are increasing despite currently available therapeutic modalities. Systemic administration of interleukin (IL)-12 has been shown to inhibit airway reactivity in murine models of asthma, but the required dosage is high and may be toxic. This study tested the hypothesis that IL-12 administered directly into the lungs is more effective in inhibiting airway reactivity than systemically administered IL-12, allowing lower doses to be used. A low dose (10 ng) of IL-12 was delivered either intratracheally (mucosal delivery) or intraperitoneally (systemic delivery) at the time of ragweed (RW) challenge in mice sensitized to RW. Basal airway resistance and airway reactivity to methacholine were measured 3 days after RW challenge. Compared to phosphate-buffered saline (PBS) challenge of RW sensitized mice, RW challenge increased basal resistance and the slope of the methacholine dose-response curve. Methacholine challenge of RW-challenged mice also induced premature respiratory failure (respiratory rate < 150/min, tidal volume < 0.15 mL) in some animals. Administration of mucosal or systemic IL-12 at the time of RW challenge decreased basal airway resistance. However, only mucosal IL-12 decreased airway reactivity and inhibited respiratory failure during methacholine challenge. These findings indicate that mucosal delivery of a low dose of IL-12 is more effective than systemic IL-12 in inhibiting airway reactivity and respiratory failure in a mouse model of asthma. PMID- 11033770 TI - [The role of genetic instability in cell aging]. AB - The current research literature on the mechanisms responsible for maintenance of the genomic stability and their role in cell senescence both in vivo and in vitro is reviewed. Various types of age-dependent genomic destabilization in senescent cells are considered. Genetic instability of senescent cells is assumed to be associated with the life span and aging of an entire organism. PMID- 11033771 TI - [Cryptic heterokaryons in strains of Saccharomyces cerevisiae. Conditions for detecting a cryptic nucleus]. AB - A phenomenon discovered earlier, cryptic heterokaryosis in Saccharomyces yeast, has been further investigated. A phenotypically silent nucleus in a yeast cell may resume its expression after fusion with another parental cell. The resulting hybrid is capable of sporulation. By the growth of a cytoductant with an expressing nucleus from one parent and a silent nucleus from the other parent on suitable selective media, the silent nucleus can be activated. The presence of deletion or insertion mutations in several genes in YPH strains allows nuclei of the YPH type to be traced not only genetically but also by blotting. PMID- 11033772 TI - [RAD29 and RAD31--new genes from Saccharomyces cerevisiae yeasts, participating in control of DNA repair. II. Clarification of possible functions of these genes]. AB - Possible functions of previously described genes RAD29 and RAD31 involved in DNA repair were determined by analyzing the interaction between these genes and mutations in the genes of the three basic epistatic groups: RAD3 (nucleotide excision repair), RAD6 (error-prone mutagenic repair system), RAD52 (recombination repair pathway), and also the apn1 mutation that blocks the synthesis of major AP endonuclease (base excision repair). The results obtained in these studies and the estimation of the capability for excision repair of lesions induced by 8-metoxipsoralen and subsequent exposure to long-wavelength UV light in mutants for these genes led to the assumption that the RAD29 and RAD31 genes are involved in yeast DNA repair control. PMID- 11033773 TI - [Oncovirus-induced permanent genetic instability in Drosophila melanogaster]. AB - Mutant alleles of a system of genetic instability induced by oncoviral DNAs were shown to demonstrate an unstable manifestation 500 generations after their emergence. A cytogenetic analysis of oncovirus-induced unstable lines has revealed numerous chromosome rearrangements. For the Lobe alleles of this system, a specific chromosome rearrangement, Df(2L) = 35C-36B, was found on the left arm of chromosome 2. We used recessive lethal mutations involving DNA rearrangements in a successful construction of cross systems for "explosive" instability. PMID- 11033774 TI - [Evaluation of genetic variability of the Kamchatka crab Paralithodes camtschaticus (Tilesius) using allozyme markers]. AB - Interspecific genetic variation in populations of red king crab Paralithodes camtschaticus Tilesius (Litholidae, Decapoda: Crustacea) was examined using allozyme markers. The activity of 57 enzymes and the general protein presumably encoded by 92 loci was detected. The level of allozyme variability was low: the expected heterozygosity and the proportion of polymorphic loci were respectively 0.027 +/- 0.008 and 6.5%. This level of heterozygosity is three times lower than the average value for 122 crustacean species (0.082 +/- 0.007). Although genetic variants were found at 22 loci, their frequencies were generally low: only in loci 6-Pgd, Alp-1, and Pep-1 did the frequencies of the most common alleles not exceed 0.9. All polymorphic loci except one had two alleles; the exception was 6 Pgd, which had three alleles. The possible reasons for the low level of allozyme variability in red king crab are discussed. PMID- 11033775 TI - [Genetic characteristics of the wing mutation Odd(22) and its interaction with alleles of the Delta locus in Drosophila virilis]. AB - The dominant sex-linked semilethal mutation Odd22 was isolated from progeny of a dysgenic cross of Drosophila virilis lines. Flies homozygous, heterozygous, and hemizygous for Odd22 displayed multiple wing defects, including enlargements and gaps on the veins; irregularly thickened, branched, shortened, or completely reduced veins; and cuts on the wing margin. The most remarkable feature of the Odd22 expression was a combination of both an increase and a reduction of the wing vein material simultaneously present in the same wing, which is commonly associated with suppression and hyperfunction, respectively, of genes of the Notch (N) signaling system. Phenotypic analysis revealed the interaction of Odd22 with alleles of the Delta (Dl) locus, which codes for the ligand of the NOTCH receptor. Based on these data, Odd22 was assumed to directly or indirectly affect the activity of the genes involved in Dl-N signaling. PMID- 11033776 TI - [Genotype-specific changes of certain functional indicators of immunocompetent cells in male laboratory mice under conditions of pheromone stress]. AB - Mature laboratory male mice CBA, C57BL/6, BALB/c, and hybrids CBAB6F1 were exposed for 2 or 24 h to vapors of 2,5-dimethylpyrazine (DMP), which is a pheromone of the house mouse. This caused changes in the content of noradrenaline within the nerve fibers of both nasal mucosa and vascular testis tunic and an inhibition of leukocyte migration in the peripheral blood. Intrastrain distinctions were also revealed in the level of spontaneous leukocyte migration and intensity of the response to the DMP. The mechanisms underlying these effects and their possible adaptive significance are discussed. PMID- 11033777 TI - [Effect of four doses of the Su(UR)ES gene on intercalary heterochromatin in Drosophila melanogaster]. AB - Polytene chromosomes of salivary glands of various Drosophila melanogaster strains containing two doses of the normal Su(UR)ES allele have a constant set of intercalary heterochromatin (IHC) sites. Their DNA is underreplicated, which leads to breaks and ectopic contacts emerging at a certain rate. Almost no underreplication, breaks, or ectopic conjugation are present in mutants lacking the normal Su(UR)ES gene product. It could be expected that an increase in the number of the Su(UR)ES+ gene doses would, in turn, drastically increase ectopic conjugation and breakage. To test this hypothesis, a strain of D. melanogaster was obtained with two additional doses of Su(UR)ES+ introduced into its genome. The flies with four gene doses exhibited a considerable increase in ectopic conjugation: both the proportion of regions participating in conjugation and the number of chromosomes with numerous contact nodes were increased. As a result, chromosomes that were straight and well-stretched in homozygotes for the mutation in Su(UR)ES became twisted and wound and contained many loops or nodes. Many chromosomes were wound too tightly for cytological analysis. Four doses of Su(UR)ES+ considerably increased the number of weak "points." For example, the 2R chromosome has only 3 weak points in strains with two doses of Su(UR)ES+ and as many as 22 weak points in the strain with four doses. In the transgenic strain, the frequency of breaks in previously known weak points increased, and new breaks appeared in 19 additional sites. All new break points appeared in the regions that were earlier described as regions of late replication in the S phase. PMID- 11033778 TI - [Effect of fusococcin on the genome of common wheat]. AB - The effect of fusicoccin on the common wheat genome (cv. Mironovskaya 808) during early phases of seed germination was studied. It was shown that fusicoccin: (1) increased the number of cells with one or two nucleoli and decreased the proportion of cells containing three and four nucleoli; (2) enhanced the total volume of nucleoli per nucleus at a concentration of 0.68 mg/l; (3) did not activate additional rRNA genes on chromosomes 1B and 6B; (4) did not activate the latent nucleolus organizer regions on chromosomes of genome B; (5) induced associations of nucleolus organizer regions on B-genome chromosomes; (6) significantly enhanced mitotic activity in apical meristem of seedling root tips and promoted the first peak of mitoses; the effect disappeared after 44 h of seed imbibition; (7) in contrast to gibberellin, did not synchronize cell division; (8) did not cause chromosome and chromatid aberrations and increase the frequency of sister chromatid exchanges at physiologically active concentrations. Presumable mechanisms of fusicoccin-induced activation of rRNA genes are discussed in relation to the identification of the fusicoccin receptor belonging to the GF14 proteins with an intracellular binding site, to the detection of endogenous fusicoccin in plants in vivo, and to fusicoccin-induced activation of gene transcription. PMID- 11033779 TI - [RAPD-PCR analysis of the variability of spring common wheat cultivar genomes and their androclinal double haploid form]. AB - Based on RAPD-PCR analysis with 15 primers including those homologous to particular loci, the level of genetic polymorphism in the collection of spring common wheat genotypes and their androclinal double haploid (ADH) lines was determined. Intraspecific polymorphism of the original wheat forms was 20%. By the absence of polymorphism in the molecular patterns of ADH lines and original forms, it was shown that, under the conditions used, no genetic changes in the genomic DNA of spring common wheat in anther culture occurred. Thus, the technology of direct in vitro androgenesis does not lead to genome rearrangements and may be used for rapid production of pure lines of such a complex allopolyploid as spring common wheat. PMID- 11033780 TI - [Absence of heterosis in hybrid corn. Description of the effect]. AB - Three unrelated homozygous maize lines, TS11, P22, and ST156, that produced hybrids in which heterosis was either absent or insignificant were identified. These hybrids were phenotypically similar to self-pollinated homozygous lines. Reciprocal crosses showed that the absence of heterosis is controlled by nuclear genes and is not associated with the cytoplasm of inbred lines. Analysis of F2 plants demonstrated that lines TS11, P22, and ST156 contained allelic genes determining the absence of heterosis in hybrid plants. Crosses of lines TS11, P22, and ST156 with a common selection line 092 generated hybrids with normal heterosis. It was concluded that heterozygosity or homozygosity of particular genes in lines TS11, P22, and ST156 play a pivotal role in the manifestation or the absence of hybrid vigor in hybrids. PMID- 11033781 TI - [Change in the spectra of peroxidases in regenerants of Stachys sieboldii (Miq.) as a result of hormonal and mutagenic effects]. AB - Nodular segments of the stem and stolons of aseptic Stachys sieboldii (Miq.) plants were cultured in Gamborg's B5 medium at a low (1-2 mg/l) or high (10 mg/l) concentration of cytokinins (6-BAP, kinetin) and produced many adventitious buds and shoots without callus formation. Several plants were regenerated from nodular segments incubated at various concentrations of 5-bromodeoxiuridine (5-BdU) for three weeks. All regenerants were morphologically normal. Peroxidase electrophoretic patterns were analyzed to determine whether somaclonal variation is associated with S. sieboldii micropropagation. The frequency of changed forms was higher with the high 6-BAP concentration and with mutagenic treatment. Changed forms were not detected in plants grown at a low cytokinin concentration, which can be used in S. sieboldii micropropagation. PMID- 11033782 TI - [Population-genetic structure of Dolly varden (Salvelinus malma Walbaum) from Southeastern Sakhalin and Southern Kuril islands]. AB - The genetic structure of Dolly Varden (Salvelinus malma Walbaum) populations from six rivers of southeastern Sakhalin and six rivers of the Kuril Islands was examined using electrophoretic, analysis of 22 enzyme systems encoded by 45 loci. In all populations, mean heterozygosities (0.044-0.105), mean numbers of alleles per locus (1.15-1.52), and proportions of polymorphic loci (18.2-39.4%) were estimated for all loci that had clear resolution. Highly significant heterogeneity of allele frequencies was detected among populations of both Sakhalin and the Kuril islands. By means of cluster analysis and multidimentional scaling, all populations were divided into the Sakhalin and Kuril groups. Within the groups, no association of genetic similarity with geographic distance between populations was observed. Although the average genetic diversity was similar in these regions, the relative interpopulation diversity was almost three times higher in the Kuril (17.6%) than in the Sakhalin (6.5%) groups. In all, the proportion of genetic diversity between regions (8.4%) was somewhat lower than the proportion of mean interpopulation diversity (11.3%). In small populations isolated from the sea, genetic variation was lower than in nonisolated populations. Allozyme differences between the northern and the southern forms of Dolly Varden are discussed. PMID- 11033783 TI - [Phylogeographic carrion, hooded and jungle crows (Aves, Corvidae) from data on partial sequencing of the mitochondrial DNA cytochrome B gene]. AB - Distribution of mitochondrial DNA cytochrome b gene haplotypes in two crow species was examined by means of sequencing of the 336-bp gene fragment. The topology of the NJ and UPGMA trees showed that the carrion crow range was split into two parts due to the presence of significantly diverged ancestral lineage localized in the southeastern part of the range. The carrion crow populations, inhabiting a territory ranging from France to northern Sakhalin, along with interspersed hooded crow populations and hybrid Siberian populations, shared a common haplotype. The border between two carrion crow lineages revealed is located in central Sakhalin. The subdivision of two weakly differentiated lineages within the jungle crown range, also observed within this territory, coincided with the subspecies division of this species. The estimated genetic distances indicate the isolation of the subgenus Coloeus. These data also suggest the convergent similarity between the chough Pyrrhocorax pyrrhocorax and the Corvus genus, as well as the conspecificity of Corvus corone corone and C. c. cornix. PMID- 11033785 TI - [Genogeographic analysis of a subdivided population. II. Geography of random inbreeding (from frequency of surnames in Adygs)]. AB - An important characteristic of the genetic structure of populations, random inbreeding (interpopulation variation), was evaluated on the basis of quasi genetic markers (surnames). The following methodological issues are considered: estimation of random inbreeding using the coefficient of isonymy fr in a subdivided population; a comparison of inbreeding levels calculated on the basis of surname frequencies using fr and Wright's FST; a comparison of inbreeding estimates obtained on the basis of surnames and genetic markers; inbreeding variation in populations of the same hierarchical rank; and planning of genetic studies of a subdivided population. The population of Adygs (an indigenous ethnic group of Northern Caucasus) was examined as a model subdivided population. The population system of Adygs is hierarchical. Parameters of random inbreeding were examined at each level of the system "ethnic group==>tribe==>geographic group of auls==>aul." Frequencies of surnames were collected subtotally. Data on frequencies of 1340 surnames in 61 auls representing all Adyg tribes were analyzed. In total, 60,000 people were examined. The inbreeding estimates obtained on the basis of Wright's FST and the coefficient of isonymy fr virtually coincided: for Adygs in general, FST x 10(2) = 2.13 and fr x 10(2) = 2.09. At the same time, the inbreeding level exhibited marked differences among tribes: in Shapsugs, these differences were an order of magnitude higher than in Kabardins (fr x 10(2) = 2.53 and 0.25, respectively). The inbreeding estimates for auls differed by two orders of magnitudes: fr x 10(2) = 0.07 and fr x 10(2) = 7.88. An analysis of ten auls yielded fully coinciding inbreeding estimates based on quasi genetic (fr x 10(2) = 0.60) and classical (FST x 10(2) = 0.69) gene markers. Computer maps of surname distributions in Adygs (1340 maps) were constructed for the first time ever. Based on these maps, the map of random inbreeding in the Adyg population was obtained. PMID- 11033784 TI - [Modeling heterochromatin regions in transgenic mice]. AB - Transgenic mice carrying bovine satellite DNA IV were obtained. The size of the transgene integrated into the mouse genome was approximately 390 kb (about 100 transgene copies) as determined by a semiquantitative PCR. Restriction analysis with isoschizomeric restrictases HpaII and MspI, showed that the alien DNA was methylated. In the genome of a transgenic founder male, two integration sites for satellite DNA IV were revealed by in situ hybridization and in situ PCR. These sites are situated on two different chromosomes: in pericentromeric heterochromatin and within a chromosomal arm. In transgenic mice, de novo formation of heterochromatin regions (C-block and the CMA3 disk within the centromeric heterochromatin of another chromosome) was revealed by C-banding and staining with chromomycin A3. This formation is not characteristic of mice, because their chromosomes normally contain no interstitial C-blocks or sequences intensely stained by chromomycin A3. PMID- 11033786 TI - [Effects of human immunodeficiency virus type 1 nef and tat genes on rat PC12 pheochromocytoma cells]. AB - The regulatory genes nef and tat of the human immunodeficiency virus type 1 (HIV 1) were transferred into the rat pheochromocytoma cells (line PC12) under the control of the eukaryotic promoters. Proliferative activity of the PC12 cells transfected with the tat HIV-1 gene was substantially increased as compared to the control. Conversely, the nef gene introduced into the cultivated PC12 cell caused inhibition of their proliferative activity and formation of cell agglomerates resembling in morphology the multinuclear syncytial cells. Thus, our results suggest that the tat gene activates proliferation of the cultivated PC12 cells, whereas the nef gene inhibits proliferation of the same cells. We have obtained for the first time a direct indication for the possible role of the nef gene in formation of multinuclear T-lymphocyte and macrophage syncytium in HIV-1 infected patients. The HIV-1 nef and tat genes had no significant effect on the neuronal differentiation of the PC12 cells induced by the nerve growth factor (NGF). PMID- 11033787 TI - [Analysis of the spectra of mutational damage of the 21-hydroxylase gene in patients with adreno-genital syndrome]. AB - The spectrum of mutations in the steroid 21-hydroxylase gene (CYP21B) and the frequency of 11 mutations among 66 patients with different forms of congenital adrenal hyperplasia (CAH) were analyzed by means of PCR amplification. Each of the CAH forms was characterized by specific spectrum of diagnostically important mutations. The salt-losing (SL) form of the disease was most frequently associated with gene deletion (39%) and the 668-13C-G mutation in the second intron (23.5%), whereas the majority of simple virilizing (SV) CAH cases were associated with the 1172N mutation in exon 4 (22%), gene deletion (16.5%), and the 668-13C-G mutation (16.5%). Mutations in the steroid 21-hydroxylase gene were detected in 70% of the chromosomes from the patients with the SL and SV forms of CAH, and only in 1.3% of the chromosomes from the patients with the nonclassic (NC) form. A total of 78 mutant chromosomes from the NC CAH patients were examined, and only one case of a gene deletion in the heterozygous state was revealed. In the individuals examined, the V281L and P30L mutations described in the NC CAH patients from other populations were not detected. This result can be explained either by the fact that NC CAH cases in Russia are associated with other major mutations, or by difficult clinical diagnosis questionable CAH cases. PMID- 11033789 TI - [Genetically dispersed effective population size determined by mitochondrial genes]. AB - A formula for variance effective population size (Ne) for analysis of mitochondrial genes is deduced and discussed. Only the female part of the population is taken into account; hence, Ne is, in the given case, the effective number of females. Ne = m2(N - 1)/sigma 2, where N is the number of females, sigma 2 is the variance of the reproductive contributions of individual females (measured as the number of their daughters that are part of the next generation), and m is the mean number of daughters per female. PMID- 11033788 TI - [Polymorphism of microsatellite loci DYS19, DYS393, and frequency of the T-C transition of the RBF5 locus on the Y-chromosome in inhabitants of the Volga-Ural region]. AB - Polymorphism of the DYS19 and DYS393 microsatellite loci and T-C transition at the RBF5 locus of the Y chromosome were analyzed in Volga-Ural populations of Bashkirs, Tatars, Chuvashes, Maris, Mordovians, Udmurts, and Komis. For the DYS19 locus, statistically significant differences were observed between Trans-Ural and Northeastern Bashkirs; between Trans-Ural Bashkirs and Tatars; and between Udmurts and other populations of the Volga-Ural region, excluding Trans-Ural Bashkirs. The DYS393 locus allele frequency distribution patterns were similar in all populations studied. The highest and the lowest frequencies of T-C transition at the RBF5 locus was detected in Udmurts (0.68) and in Mordovians (0.09), respectively. Association of C-alleles with the DYS19/DYS393 microsatellite haplotypes was investigated. The major haplotypes specific to the Turkic- and Finno-Ugric populations were revealed. PMID- 11033790 TI - [Molecular-genetic analysis of VNTR polymorphism in alleles of the phenylalanine hydroxylase gene in inhabitants of the Volga-Urals region]. AB - Molecular genetic analysis of the VNTR alleles at the phenylalanine hydroxylase (PAH) gene was carried out in seven Volga-Ural ethnic groups (Bashkirs, Tatars, Chuvashes, Maris, Mordovians, Udmurts, and Komis). The PCR fragments revealed included alleles of 380, 440, 470, 500, 530, 560, and 650 bp, containing 3, 5, 6, 7, 8, 9, and 12 repeat copies, respectively. Substantial heterogeneity of the populations in respect to the distribution and frequency of the VNTR alleles and genotypes was demonstrated. The indices of observed and theoretical heterozygosity of the PAH VNTR alleles were calculated. The mean heterozygosity index was 70.02%. This high index value along with the established differentiation of the populations in respect to the frequency distribution of the VNTR alleles and PAH genotypes permitted the conclusion that the given polymorphic locus can serve as a highly informative marker for examination of the genetic structure of the Volga-Ural populations. PMID- 11033792 TI - [13th Annual meeting of the Retinology Society. 23-24 June 2000, Freiburg. Abstracts]. PMID- 11033791 TI - [Study of the phenotypic occurrence of ura gene inactivation in Bacillus subtilis]. AB - After inactivation of the ypaA gene in Bacillus subtilis, the phenotypic pattern obtained showed that this gene controls a system for active flavin transport and, possibly, riboflavin excretion under the conditions of constitutive synthesis. PMID- 11033793 TI - [In Process Citation] PMID- 11033794 TI - [Mts1 protein and control of tumor metastasis]. PMID- 11033795 TI - [Mixed culture of eukaryotic cells: cytotoxic processes]. PMID- 11033796 TI - [Molecular genetic mechanisms for regulating the tissue-specific expression of the estS gene in drosophila]. PMID- 11033797 TI - [Different mechanisms of regulating telomere length]. PMID- 11033798 TI - [Conserved and variable nature of eukaryotic ribosomal DNA]. PMID- 11033799 TI - [Paracrine regulation of the c-met proto-oncogene transcript by stromal cells in BMP tumor cells with different metastatic potential]. PMID- 11033800 TI - [Millimolar concentration of methylmethanesulfonate increases the level of transcription with the CMV promoter by more than 100 times]. PMID- 11033801 TI - [A high constitutive level of NF-kappa B is necessary for viability of murine adenocarcinoma cells--possible role of p53]. PMID- 11033802 TI - [Molecular characteristics of two new homologs of human TAFII-30 in Drosophila melanogaster]. PMID- 11033803 TI - [Study of e(y)1/TAFII40 gene in Drosophila melanogaster in vivo]. PMID- 11033804 TI - [Comparison of the structure and embryonal expression of the Delta locus in Drosophila virilis and Drosophila melanogaster]. PMID- 11033805 TI - [Amplification of DNA occurs between the core sequence of the replication of origin ori+ of the ts plasmid integrated in the chromosome, and its analog in the genome of the bacterial cell]. PMID- 11033806 TI - [Overexpression of the apc10+ gene in the fission yeast Schizosaccharomyces pombe can suppress temperature sensitivity of the nuc2-663 mutant,but not its sterility]. PMID- 11033807 TI - [Molecular mechanisms of cytolytic processes induced by human LAK cells]. PMID- 11033808 TI - [Regulatory sequences of DNA, located in segments hypersensitive to DNAase I in the 5-terminal region of chicken alpha-globin genes]. PMID- 11033809 TI - [Cloning and characteristics of RAPD-markers from the genome of the parasitic nematodes Trichinella spiralis and Trichinella pseudospiralis]. PMID- 11033810 TI - [DNA fingerprinting of parthenogenetic families of Lacerta-species lizards: detection of genetically unstable loci]. PMID- 11033811 TI - [Tritium planigraphy as an instrument for studying the spatial structure of proteins and their complexes]. PMID- 11033812 TI - [Antirestriction]. PMID- 11033813 TI - [Nucleotide substitution C1167T in the catalase gene and position of nearby polymorphic markers DS11S907 and D11S2008 are connected with development of diabetes mellitus type 2]. PMID- 11033814 TI - [Study of local correlations in bacterial genes for use in frames of hidden Markov models]. PMID- 11033815 TI - [Polymorphic minisatellite of ecNOS4a/4b in the endothelial NO-synthase gene amd cardiovascular diseases]. PMID- 11033816 TI - [Mechanism of oligonucleotide hybridization with the 3'-terminal region of yeast tRNA(Phe)]. PMID- 11033817 TI - [Dissociation of proteins from the 40S subunit of human ribosomes in a lithium chloride gradient]. PMID- 11033818 TI - [An analog of mRNA--a derivative of pUUUGUU with an arylazide group at the guanosine residue, crosslinks with nucleotides A-1823 and A-1824 of rRNA within the human 80S ribosome]. PMID- 11033819 TI - [Mutation of a glutamine residue in the universal tripeptide GGQ in human eRF1 termination factor does not cause complete loss of its activity]. PMID- 11033820 TI - [Catalytic effectiveness of subunits of a dimeric molecule of aminotransferase in solution and in crystalline form]. PMID- 11033822 TI - [In Process Citation] PMID- 11033821 TI - [Vesicular transport gene bet1 is differentially transcribed in transformed cells with various metastatic potential]. PMID- 11033823 TI - Optimal use of the newer antifungal agents. PMID- 11033824 TI - Update on adjuvant chemotherapy for early breast cancer. AB - Adjuvant chemotherapy represents a significant advance in the management of early stage breast cancer and, as such, has saved many lives. Worldwide, adjuvant chemotherapy has benefitted all groups tested, including pre- and postmenopausal women, those with node-negative and node-positive disease, and those with estrogen-receptor (ER)-positive and ER-negative disease. However, the significant number of women who relapse despite adjuvant therapy provides the impetus to develop more efficacious regimens. Results from large randomized clinical trials, which will mature over the next few years, will clarify the potential benefits of the taxanes in the adjuvant setting, provide answers as to the efficacy of a dose dense approach, and define a role, if any, for high-dose chemotherapy. A shift toward targeted therapies has also begun, with the incorporation of trastuzumab (Herceptin) into the adjuvant setting. Minimizing the long-term toxicity of adjuvant therapy for the large number of women who survive their disease is paramount. This article highlights the need to develop predictive factors to help tailor individual therapy. PMID- 11033825 TI - New drug delivery system may offer clinical advantages in prostate cancer treatment. PMID- 11033826 TI - Longer time to tumor progression with exemestane vs tamoxifen in advanced breast cancer. PMID- 11033827 TI - Camptosar plus cisplatin increase survival in small-cell lung cancer study. PMID- 11033828 TI - Pharmacology and clinical status of capecitabine. AB - Capecitabine (Xeloda) is a new, orally administered, enzyme-activated fluoropyrimidine carbamate designed to generate high levels of fluorouracil (5 FU) in tumor cells. Selective tumor activation of 5'-deoxy-5-fluorouridine, the last enzymatic step of a three-enzyme process, is catalyzed by thymidine phosphorylase, a tumor-associated angiogenic growth factor. Since levels of thymidine phosphorylase are often higher in tumors than in surrounding normal tissues, this stepwise process provides tumor selectivity and potentially decreases toxicity to normal tissues. Preclinical studies show that capecitabine has significant activity against a variety of tumor types when used as monotherapy and in combination with other chemotherapeutic agents. Capecitabine is currently approved for the treatment of patients with metastatic breast cancer resistant to paclitaxel (Taxol) and in whom further anthracycline therapy is contraindicated. It has demonstrated efficacy in patients with advanced colorectal carcinoma, and studies are ongoing in other tumor types. PMID- 11033829 TI - Clinical trials referral resource. Current clinical trials administering the antiangiogenesis agent SU5416. PMID- 11033830 TI - Recent developments in chemoradiotherapy for locally advanced cancer of the cervix. AB - Patients with locally advanced cervical cancer comprise a significant proportion of the total population with cervical cancer, particularly in developing countries. The inability to control pelvic tumors is still a significant concern. Although neoadjuvant chemotherapy is associated with a high response rate, data from randomized trials clearly do not support the use of neoadjuvant chemotherapy prior to definitive irradiation. However, the results of concurrent cisplatin (Platinol)-based chemotherapy and radiotherapy are highly promising for locally advanced cancer of the cervix and should be considered as a treatment option. To decrease the risk of distant metastasis and improve survival, more effective drugs or drug combinations need to be developed. PMID- 11033831 TI - Cancer, unproven therapies, and magic. AB - Commonly used by cancer patients, unproven therapies are treatments that the practitioner claims can alter the disease process although there is no proof to support the claim. The reasons for the popularity of unproven therapies fall into two categories--practical considerations and fundamental mechanisms. Research has implicated the following practical factors: a pragmatic search for relief of symptoms, expression of a philosophical view, a need to reestablish a sense of control in life, and dissatisfaction with conventional medicine. Fundamental mechanisms include traditional magic, the heroic individual, and a delusional pattern of thinking. Underpinning and generating these factors is the fear of death. Particularly in patients with cancer, this is not only a fear of nonexistence, but of loneliness, the unknown, pain, loss of control, and emptiness. The popularity of unproven therapies poses a challenge to the medical system at large, and oncologists, psycho-oncologists, and palliative-care physicians, in particular. The essence of the challenge is to understand the reasons for the use of unproven therapies, to analyze our own behavior, and conclude what if anything our response should be. Unproven therapies (as with magic, a sense of heroism, and delusional thinking) fulfill the function of resolving the inexplicable and the psychologically painful--i.e., relieving the anxiety associated with cancer. PMID- 11033832 TI - Use of bisphosphonates in the treatment of prostate cancer. PMID- 11033833 TI - rhIL-11 for the prevention of dose-limiting chemotherapy-induced thrombocytopenia. PMID- 11033834 TI - Preclinical pharmacologic basis for clinical use of rhIL-11 as an effective platelet-support agent. AB - Preclinical studies have shown that rhIL-11, also known as oprelvekin (Neumega), stimulates early and later stages of megakaryocytopoiesis (including proliferation and differentiation of megakaryocyte precursors and maturation of megakaryocytes), to produce an increase in peripheral platelet count. Because of these effects, rhIL-11 must be administered to patients with cancer sufficiently in advance of the platelet nadir (within 6 to 24 hours postchemotherapy) to allow adequate time for megakaryocyte maturation and platelet formation. Therefore, the maximum platelet response coincides with the time when the platelet nadir would normally be experienced. In myelosuppressed, nonhuman primates, optimal platelet response occurred following 14 days of treatment at a dose equivalent to the 50 microgram/kg daily dose recommended in humans; lower doses and shorter durations were less effective. These data support the current dosing recommendation in humans, which states that rhIL-11 dosing continues until platelet recovery to > or = 50,000/microL has been achieved for 2 consecutive days or for a total of 10 to 21 days in each cycle. The nonhematopoietic effects of rhIL-11 include a renal effect, resulting in plasma-volume expansion, as well as potential beneficial clinical effects in damaged or inflamed intestinal mucosa, including potential mitigation of mucositis and a rationale for future studies in inflammatory bowel disease. PMID- 11033835 TI - Dose reductions and delays: limitations of myelosuppressive chemotherapy. AB - Thrombocytopenia occurs at various grades of severity in patients with nonmyeloid malignancies undergoing chemotherapy with myelosuppressive agents. Frequently, it is the major dose-limiting hematologic toxicity, especially in the treatment of potentially curable malignancies such as leukemia, lymphomas, and pediatric cancers. This is becoming increasingly important given the recent trend toward the use of dose-intensive combination chemotherapy regimens facilitated by supportive hematopoietic colony-stimulating factors to prevent chemotherapy induced febrile neutropenia. The standard preventive measure against chemotherapy induced depression of platelets in subsequent treatment cycles has been dose reduction and/or dose delay. However, follow-up data from studies in various populations of patients with cancer suggest a correlation between delivery of lower than intended doses and poor outcomes, including reduced disease-free periods and overall survival. Other consequences of thrombocytopenia include the need for platelet transfusions and subsequent exposure to the risk of numerous complications, including bacterial and viral infections; febrile, nonhemolytic transfusion reactions; and transfusion-induced immunosuppression. Furthermore, a large proportion of multitransfused patients become refractory to subsequent infusions. Refractoriness to platelet transfusions is quickly becoming more prominent. The availability of a platelet growth factor--recombinant human interleukin-11(rhIL-11, also known as oprelvekin [Neumega])--provides an effective means of preventing chemotherapy-induced thrombocytopenia and accelerating platelet recovery, thereby facilitating the administration of full doses of chemotherapy during subsequent cycles and avoiding the need for rescue with platelet transfusions. PMID- 11033836 TI - Clinical efficacy of rhIL-11. AB - Placebo-controlled clinical trials of recombinant human interleukin-11 (rhIL-11, also known as oprelvekin [Neumega]) in patients with nonmyeloid malignancies have demonstrated significant efficacy in preventing postchemotherapy platelet nadirs of < or = 20,000/microL, and reducing the need for platelet transfusions while continuing chemotherapy without dose reductions. The likelihood of requiring a platelet transfusion in rhIL-11-treated patients receiving chemotherapy is approximately 40% lower than the risk for untreated patients. Treatment with rhIL 11 appears to accelerate earlier recovery to platelet counts of 20,000/microL, 50,000/microL, and 100,000/microL, suggesting that patients treated with rhIL-11 are more likely to be able to receive their next chemotherapy cycle in a timely fashion. rhIL-11 shows sustained efficacy over multiple cycles of full-dose chemotherapy. Activity has also been demonstrated in pediatric patients with solid tumors. The use of rhIL-11 in combination with granulocyte colony stimulating factor (G-CSF, filgrastim [Neupogen]) may also produce a synergistic hematopoietic effect, resulting in earlier neutrophil recovery. The recommended adult dose regimen for rhIL-11 is 50 micrograms/kg administered subcutaneously once daily beginning 6 to 24 hours after the administration of chemotherapy until a postnadir platelet count of > or = 50,000/microL is reached. The recommended pediatric dose of rhIL-11 is 75 micrograms/kg subcutaneously, once daily beginning 6 to 24 hours after the administration of chemotherapy until a postnadir platelet count of > or = 50,000/microL is reached. The administration of rhIL-11 for greater than 21 days has not been studied and therefore is not recommended. PMID- 11033837 TI - Tolerability and side-effect profile of rhIL-11. AB - Safety data from two randomized phase II and one abbreviated phase III placebo controlled, double-blind clinical studies in adult patients with nonmyeloid malignancies indicate that recombinant human interleukin-11 (rhIL-11, also known as oprelvekin [Neumega]) has an acceptable toxicity profile as therapy for the mitigation of chemotherapy-induced thrombocytopenia. Preliminary data also indicate that rhIL-11 is well tolerated by pediatric patients with similar types of cancers. Adverse events associated with rhIL-11 are generally mild or moderate, reversible with drug discontinuation, and easily managed. Many of the common adverse events of rhIL-11--including edema, dyspnea, pleural effusions, conjunctival injection, and in some patients, atrial arrhythmia--occur in association with fluid retention. However, these adverse events can be medically managed and need not limit the use of rhIL-11, particularly if ameliorative measures, such as salt restriction and occasional prophylaxis with a potassium sparing diuretic to minimize peripheral edema, have been instituted along with close monitoring of fluid and electrolyte status. Such measures are suggested for any patient treated with a diuretic, especially patients with cancer who are receiving multiple medications that complicate overall care. Administration of sequential cycles of rhIL-11 treatment does not appear to result in an increased incidence of adverse events or bone marrow exhaustion. rhIL-11 does not appear to interact adversely with concomitantly administered chemotherapeutic agents or agents commonly used for supportive care, including granulocyte colony stimulating factor (G-CSF, filgrastim [Neu-pogen]). PMID- 11033838 TI - Facts and controversies in the classification of idiopathic interstitial pneumonias. AB - Idiopathic interstitial pneumonias are defined from the pathological point of view as non granulomatous intralobular inflammatory and fibrotic processes involving the alveolar walls. More than thirty years ago Liebow and Carrington pioneered the notion that morphological characteristics could be used with benefit in separating the different entities found in this group, which present with typical, but not pathognomonic clinical features. In the mid-1980s some entities, including giant cell interstitial pneumonia (GIP) and lymphocytic interstitial pneumonia (LIP), were removed from this group and considered as peculiar forms. In the early 90s the concept of cellular or nonspecific interstitial pneumonia was reconsidered, leading to an in depth revision of various types of interstitial pneumonia of unknown etiology. The histological pattern observed in patients with idiopathic pulmonary fibrosis is now referred to as usual interstitial pneumonia (UIP). Other entities that have been revised during the last ten years are desquamative interstitial pneumonia/alveolar macrophage pneumonia (DIP/AMP), respiratory bronchiolitis-interstitial lung disease (RB-ILD), acute interstitial pneumonia (AIP), cryptogenic organizing pneumonia (COP) and nonspecific interstitial pneumonia (NSIP). This paper provides a detailed description of pulmonary disorders which have been included in the new classification systems of idiopathic interstitial pneumonias. In the second part of the paper we will discuss several doubts and controversies that this new classification schemes leave unresolved. PMID- 11033839 TI - Structural basis for airflow limitation in chronic obstructive pulmonary disease. AB - The airflow limitation that characterises chronic obstructive pulmonary disease (COPD) has two main components: an increased resistance, which is due to airway obstruction, and a loss of the elastic recoil pressure of the lung, which is due to parenchymal destruction. Although it has long been known that the major site of increased resistance in COPD is the peripheral airways, recent studies have shown that central airways are involved in the disease as well. The purpose of this review is to describe the major structural and cellular changes present in peripheral airways, central airways and lung parenchyma of patients with COPD, and to underline the possible mechanisms contributing to airflow limitation in these subjects. PMID- 11033840 TI - Antigen presentation in the lung: dendritic cells and macrophages. AB - Antigen presentation is a required prime event before T-cell activation can occur. Cells which constitutively express major histocompatibility antigen class I or II are responsible for presenting antigens. These are essentially alveolar macrophages (AM) residing mostly in the air spaces, and dendritic cells (DC), which create a tight surveillance network just below the epithelial cells of the airways and in the loose connective tissue around the vessels or in the pleura. AM are poor antigen presenting cells compared to DC. AM when encountering foreign particles or organisms may, however, influence the degree of activity or maturation of neighbouring DC, by releasing cytokines. Thus, we will describe how the innate immune processes may influence specific immunity and perhaps Th1 and Th2 differentiation. Following the description of the differences in phenotype and functions of AM and DC, we will provide data showing that in some pathological conditions, such as sarcoidosis, AM can acquire some specificities of DC. PMID- 11033841 TI - Proliferative response of peripheral blood mononuclear cells and levels of antibody to recombinant protein from Propionibacterium acnes DNA expression library in Japanese patients with sarcoidosis. AB - BACKGROUND AND AIM OF THE WORK: The causes of sarcoidosis are unknown. Propionibacterium acnes has been isolated from sarcoid lesions, and many genomes of P. acnes or P. granulosum have been detected in all biopsy samples tested from Japanese patients with sarcoidosis. We searched for protein antigens from propionibacteria that caused immune responses in patients with sarcoidosis but not in subjects without sarcoidosis. METHODS: A lambda gt11 genomic DNA expression library of P. acnes was screened with sera from patients with sarcoidosis. Antibodies to a recombinant protein from the insert recovered by the screening were measured in serum and bronchoalveolar lavage (BAL) fluid from patients with or without sarcoidosis by an immunofluorescence-based method. Peripheral blood mononuclear cells from patients with and without sarcoidosis were used to examine the lymphoproliferative response to the protein. RESULTS: Of 180,000 plaques screened, two clones coded for an identical recombinant protein, termed RP35, were recognized by sera. RP35 was the C-terminal region of P. acnes trigger factor. RP35 caused sarcoidosis specific proliferation of the mononuclear cells from 9 (18%) of the 50 patients with sarcoidosis; in a similar way, purified protein derived from Mycobacterium tuberculosis evoked specific responses in 8 (38%) of 21 patients with tuberculosis. Serum levels of IgG and IgA antibodies to RP35 were high in patients with sarcoidosis and other lung diseases. In BAL fluid levels IgG or IgA antibodies were high in 7 (18%) and 15 (39%), respectively, of 38 patients with sarcoidosis, and in 2 (3%) and 2 (3%), respectively, of 63 patients with other lung diseases. CONCLUSIONS: The RP35 protein from P. acnes causes a cellular immune response in some patients with sarcoidosis but not in subjects without sarcoidosis. PMID- 11033842 TI - Vitamin D receptor gene polymorphism and calcium metabolism in sarcoidosis patients. AB - BACKGROUND: Hypercalcemia has been recognized as an important complication of sarcoidosis, caused by overproduction of the active form of vitamin D, 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) at sites of granulomatous reactions. Polymorphism of the vitamin D receptor (VDR) gene has recently been shown to be related to bone mineral density, and also associated with hyperparathyroidism and risk of granulomatous disease. In light of the possible impact on hypercalcemia of sarcoidosis, an investigation of calcium metabolism and polymorphism of the VDR gene in sarcoidosis patients was carried out. METHODS: Genotypes were determined using the polymerase chain reaction and restriction fragment length polymorphism. Maximum calcium, 1,25(OH)2D3, and intact PTH levels were also determined. RESULTS: Depressed PTH levels were found in sarcoidosis patients, especially in those with the bb genotype, but there was no difference in 1,25(OH)2D3 levels among the VDR genotypes, and this polymorphism also had no association with onset of hypercalcemia. CONCLUSION: From these results, we speculate that although the VDR gene polymorphism may affect the serum PTH level, it is not a risk factor for hypercalcemia in sarcoidosis. PMID- 11033843 TI - The value of interleukin-12 as an activity marker of pulmonary sarcoidosis. AB - BACKGROUND: Sarcoidosis is characterized by hyperactivity of T-helper lymphocytes and recent studies showed that they were mainly Th1 cells. IL-12 is a major cytokine inducing Th1 differentiation of naive T cells. This study was performed to test whether IL-12 can be a marker for disease activity and possibly a prognosis in sarcoidosis. METHODS: IL-12 levels of BALF (BALF-IL-12) and conditioned medium of alveolar macrophages (AM) were measured by ELISA in 36 patients with pulmonary sarcoidosis (14 males and 22 females, mean age: 39.6 +/- 11.0 years) and eleven normal controls. Clinically, 16 patients had active sarcoidosis and 20 had an inactive disease. RESULTS: BALF-IL-12 of sarcoidosis patients (41.3 +/- 43.9 pg/ml) was significantly higher than that of normal controls (2.5 +/- 0.4 pg/ml) (p < 0.001). The patients with active disease (71.3 +/- 54.3 pg/ml) had a higher BAL-IL-12 level than those with inactive disease (17.3 +/- 13.8 pg/ml) (p = 0.0001). It had a significant correlation with the number of T4 cells (p = 0.0001), total cell number, number and percentage of lymphocytes (p = 0.0001) and AM (p = 0.001) in BALF. It was also significantly correlated with soluble ICAM-1 levels in serum (p = 0.0001) and BALF (p = 0.002), and ICAM-1 expression of AM (p = 0.001). Furthermore the patients whose condition worsened without therapy had a significantly higher initial BALF-IL-12 level than the patients whose condition improved spontaneously. The AM of sarcoidosis secreted significantly more IL-12 (133 +/- 177 pg/ml) than AM of controls (68.3 +/- 43.7 pg/ml) (p = 0.038). CONCLUSION: Our data suggest that the BALF-IL-12 level can be used as a marker of the activity of pulmonary sarcoidosis and possibly prognosis. PMID- 11033844 TI - Ketoconazole for the treatment of refractory hypercalcemic sarcoidosis. AB - Hypercalcemia is a recognised complication of sarcoidosis due to excess 1,25 hydroxyvitamin D3 production by macrophages. Systemic corticosteroids inhibit 1,25-hydroxyvitamin D3 production, but long term therapy is often required to maintain normocalcemia. Ketoconazole is an imidazole antifungal that inhibits macrophage 1 alpha-hydroxylation of 25-hydroxyvitamin D3 and has been used in paraneoplastic hypercalcemia. We report a case series of four patients with relative contraindications to corticosteroids in whom treatment with ketoconazole allowed cessation or reduction in the maintenance dose of corticosteroids. We conclude that ketoconazole should be considered as an alternative therapy for hypercalcemic sarcoidosis when corticosteroids are relatively contraindicated. PMID- 11033845 TI - One year follow-up of lung clearance of 99mTc-diethylene triamine penta-acetic acid and disease activity in sarcoidosis. AB - BACKGROUND AND AIM OF THE WORK: In interstitial lung disease lung clearance of 99mTc-diethylenetriaminepentaacetic acid (DTPA) reflects alterations in the alveolar capillary barrier. Our objective was to describe changes in lung clearance during one year in sarcoidosis, and to relate clearance to other data of lung function and disease activity. METHODS: Twenty-three newly diagnosed patients were studied with respect to lung DTPA clearance, spirometry, 67Ga scintigraphy and serum angiotensin converting enzyme (SACE). Lung mechanics and arterial PO2 at rest and exercise were studied in patients with radiological parenchymal changes. Six of these patients were prescribed peroral steroids. RESULTS: At inclusion lung DTPA clearance measured over 30 minutes was 53 +/- 16 minutes and at follow-up 59 +/- 20 minutes (p > 0.05). The number of pathological clearance curves at inclusion was 10 and at follow-up 6. In treated patients lung DTPA clearance improved as did lung 67Ga score and SACE. However, lung clearance did not correlate significantly with changes in lung function or other activity parameters. CONCLUSIONS: Lung DTPA clearance reflects lung function from an aspect different from that of other methods. It seems sensitive to pathology reversed by steroids and may, in some cases, help in the evaluation of disease activity. PMID- 11033846 TI - Eosinophilic pleural effusion due to mesalamine. Report of a rare occurrence. AB - Mesalamine-induced lung toxicity has often been described. We report on a case of a patient who underwent mesalamine treatment, though in the absence of established criteria required for diagnosing Crohn's disease (CD) or ulcerative colitis (UC). He developed an adverse respiratory reaction to the drug, thus definitely proving its lung damaging capacity. The clinical presentation included eosinophilic pleural effusion, a feature never previously described in association with mesalamine intake. PMID- 11033847 TI - Diffuse panbronchiolitis observed in a white man in Turkey. AB - Diffuse panbronchiolitis (DPB) is a distinct clinicopathological entity which is characterized by chronic recurrent sinopulmonary infection and inflammation. It is prevalent in the Far East, but extremely rare in western countries. In this report, we described the first Turkish case of DPB and also evaluated demographic and diagnostic characteristics of 18 patients with DPB, previously reported from non-Asian countries. We believe that DPB patients described from these countries may also increase if this disease is taken into consideration in differential diagnosis of sinopulmonary diseases. Another consideration is that, since the treatment and prognosis of DPB are different from other obstructive pulmonary diseases, it should not be mistaken for others. PMID- 11033848 TI - Sarcoid-like pulmonary lesions during asbestosis. A case report. PMID- 11033849 TI - [Cytogenetic disorders iin patients with chronic hepatitis B and C]. AB - The chromosomal apparatus of peripheral blood lymphocytes was studied in 19 patients with chronic hepatitis B (CHB) and 17 patients with chronic hepatitis C (CHC). Significant changes in cytogenetic indices were revealed in the both groups. Incidence of aberrant cells in CHC exceeded significantly that in CHB (15.3 +/- 1.5% vs 11.1 +/- 0.9% respectively; P < 0.05). Frequencies of aberration per cell and chromatid fragments were also significantly higher in CHC compared with both CHB and practically normal donor groups. PMID- 11033850 TI - [A molecular genetic analysis of the mutations in the exons of the CFTR gene in cystic fibrosis patients in Ukraine]. AB - The results of DNA analysis of major mutations in CFTR gene in 260 families from Ukraine with high risk of cystic fibrosis are presented. The perspectives of molecular diagnosis as a main tool for cystic fibrosis prevention are discussed. PMID- 11033851 TI - [The distribution of radiation-induced breaks in the chromosomes of irradiated subjects]. AB - Distribution of radiation-induced breakpoints in chromosomes and its bands in persons recovered from acute radiation sickness and personnel from Chernobyl NPP were investigated using G-banding staining. The frequency of damaged bands and breakpoints in groups exposed to radiation was significantly higher as compared with the control group. It was shown that in exposed to radiation persons damage depends on its length. Most frequently damaged bands in the observed groups were determined. The G-negative bands and telomeres of chromosomes were more sensitive to radiation. PMID- 11033852 TI - [A comparative analysis of plant hereditary variability during mutagenic treatment of the generative cells and seeds of corn]. AB - The effect of mutagenic factors on maize generative cells during different gametogenesis stages, as well as on mature pollen and gametes causes an essential raise in frequency, widening in visible mutation spectra and seems to be more efficient than on dry seeds. Original mutations on qualitative and quantitative characters have been obtained. PMID- 11033853 TI - [The manifestation of the determinative growth trait in the shoots of fodder beet seed plants]. AB - The peculiarities of manifestation of character "determination growth of shoots" in materials of lines of mangel beet was studied. The several types of flowering shoots with various phenotypes were picked out. Recessive nature of the character and its inheritance were established. PMID- 11033854 TI - [The genetic polymorphism of the dioeciousness trait in the fodder beet]. AB - The data on the study of genetics of the monofloret-cluster trait in fodder beet were presented. Three alleles of monofloret mc4, mc5 and mc6 were selected and described. Available monofloret sources of fertile fodder beet was tested on allelism. PMID- 11033855 TI - [The detection of nonallelic to known genes of resistance to Tilletia caries (DC) Tul. in wheat strains from interspecific hybridization (Triticum aestivum x Aegilops cylindrica)]. AB - It was established by hybridological analysis that winter bread wheat lines 1/74 91, 3/36-91, 5/55-91 possess single dominant gene of resistance to bunt (Tilletia caries (DC) Tul.), but lines 8/2-91, 5/43-91, 4/11-91 and 8/16-91 have two independent dominant genes for this character. These genes originated from Aegilops cylindrica are not identical to Bt1-Bt17 genes and are unknown to date. The lines were obtained from crosses between winter bread wheat variety Odeskaya polukarlikovaya and Aegilops cylindrica. PMID- 11033857 TI - [The morphogenetic characteristics of the gametes and the physiological features of animal reproduction in relation to their hydridization]. AB - Data of analysis of morphogenetic character of spermii in 100 species of mammals and some hybrid forms, anatomical and physiological peculiarities of reproduction of animals of various taxonomic groups, reproduction and biological suitability of generative elements of distant hybrids reflect their biological interrelation in the zoological series. It is conditioned by genetical, physiological and biochemical factors and reveals itself in different effectiveness of hybridisation processes which are determined by genealogical proximity of initial species. PMID- 11033856 TI - [The mapping of the barley genome by RAPD analysis using double haploid strains]. AB - A map of the barley genome consisting of 107 loci was constructed using double haploid lines. 33 RAPD loci marks all barley chromosomes. The total length of the obtained map is 1047 centiMorgans (cM) with a 9.8 cM average distance between markers. 7 linkage groups was identified as certain chromosomes. The distribution of the markers was analyzed, comparison with other published maps was made. Some statistical considerations are presented. Alternative strategies of gene mapping are considered. PMID- 11033858 TI - [Variability of the level of thermoresistance as an indicator of the functional status of a cell]. AB - Effect of acclimating temperature on the thermoresistance of isolated ciliated gill epithelium of Anodonta anatina was studied. One strip of gills from each mussel was used to determine the initial level of thermoresistance (survival time at 40 degrees C). Other strips were kept at 24 degrees C until their death, and their thermoresistance and fluorescence after Ethidium bromide staining were determined at different time intervals. After a short exposure at 24 degrees C the mean level of thermoresistance of the epithelium increased. The individual shift in the value and direction of cell thermoresistance in different mussels negatively correlated with its initial level. It resulted in narrowing the extent of individual variability of the level of thermoresistance. The prolongation of exposure at 24 degrees C decreased the mean level of thermoresistance, disturbed the negative correlation, and increased the variability of thermoresistance level. The value of cell fluorescence remained constant within 48 h. The increase in fluorescence was observed 12 h after reducing the mean thermoresistance level. Hence, the dynamics of variability in the thermoresistance level is a most sensitive indication of the cell functional state. PMID- 11033859 TI - [Somatic polyploidy in neurons from gastropod mollusks. I. Morphological characteristics of ganglia and neurons in the CNS of the snail Succinea lauta]. AB - A study was made of general anatomy and histological organization of the central nervous system in Succinea lauta, a gastropod pulmonate snail. Neurons grow by means of polyploidy during the postnatal succineid ontogeny. In the adult individuals diameters of perikaria of large neurons increase 2.5 to 5 times, in comparison with young individuals. As a whole, in adult snails sizes of ganglian cells vary from 3 to 380 mkm. Most of giant neurons are in parietal, abdominal and pallial ganglia. The chromatin structure has been described in neurons of young and adult snails. The endomitotic mechanism of polyploidization in the succineid ganglioneurons is proposed. PMID- 11033860 TI - [Morphofunctional study of the organization of the peripheral parenchyma of Acoela Oxyposthia praedator]. AB - The ultrastructure of undifferentiated cells in the peripheral parenchyma of Oxyposthia praedator was studied, along with the ways of their differentiation. The type I cells (3.5-4.0 microns in diameter) undergo mitotic division, while the type II cells (9 microns in diameter) produce specialized cells of the parenchyma. At the beginning of secretory cell differentiation one cistern of the rough endoplasmic reticulum (RER) is formed by the outer membrane of the nuclear envelope, the formation of other cisternae follows. The Golgi complex is formed simultaneously. The differentiated secretory cells are characterized by the abundance of RER cisternae and Golgi complexes. In the course of differentiation of other cell types RER cisternae are formed by several portions of the nuclear envelope. The Golgi complex appears in cells 12-14 microns long. The differentiation of digestive cells is characterized by autophagy. Autophagosomes are formed by RER cisternae. The consecutive stages of autophagosome formation are described. Using a cytochemical reaction revealing acid phosphatase the process of digestion of the autophagosome content was followed. PMID- 11033861 TI - [Features of the development of homo- and heterotopic allotransplants of rat embryonal neocortex]. AB - Mechanisms of regulation of cell division in the developing neocortex are largely unknown. The aim of the present study was to investigate the influence of a microenvironment on the fetal neocortex histogenesis. The fetal neocortex from 15 day old Wistar rat embryo was grafted into the neocortex, crushed sciatic nerve and anterior chamber of eye of adult rats. A comparative study of graft development was carried out on 1, 3, 7, 10, 30 days using histological (Nissl stain, hematoxylin-eosin) and immunohistochemical (monoclonal antibody to proliferating cell nuclear antigen, and to glial fibrillary acidic protein) methods. Grafted neuroepithelial cells proliferated in grafts that developed in the neocortex and the anterior chamber of eye for 7 days, and in the sciatic nerve for 10 days. In all grafts differentiating neuroblasts, young neurons and mature neurons were observed 7, 10 and 30 days later, respectively. In 10 days, transplants in the nerve have a glial capsule, in contrast to other sites of grafting. The capsule consists of ependymocytes with microvilli and cilia 30 days later. These cells are GFAP-positive. Our results indicate epigenetic influence on the development of neuroepithelial precursors. The microenvironment of the peripheral nerve is suggested to promote glyogenesis in developing grafts. Afferent inputs do not influence the proliferative potency of brain cell precursors. PMID- 11033862 TI - [Effect of "vilon" on cirrhotically changed rat liver. Liver regeneration, and status of glycogen-forming function of hepatocytes]. AB - Effects of a dipeptide preparation "Vilon" on rehabilitation of functional activity of hepatocytes and regeneration of the cirrhotically altered rat liver were studied. The liver cirrhosis was produced by poisoning of rats for 4 months with carbon tetrachloride (CCl4). On the end of the poisoning with CCl4, one group of animals was not submitted to any further actions, whereas animals of the other group were injected "Vilon" (1.7 micrograms/kg) daily for 5 days. On smears of isolated hepatocytes, contents of total glycogen (TG), and its labile and stable fractions (LF and SF) were determined in addition to cell ploidy levels and the total protein content. In liver homogenates, activities of glucose-6 phosphatase (G6P), glycogen synthase (GS), and glycogen phosphorylase (GP) were measured. In 2 weeks after the drug application, G6P activity being reduced in cirrhosis 1.2 times, elevated under effect of "Vilon". In non-treated rats the contents of TG and its fractions and of G6P activity remained at the level characteristic of the cirrhotic liver prior to "Vilon" administration. In both groups of rats, GP and GS activities in the cirrhotically altered liver did not differ from their control values throughout the entire experiment. "Vilon" has been shown to exert a weak stimulating effect on regeneration of the cirrotically altered rat liver: in hepatocytes of the second group of rats the total protein content and ploidy levels were higher than those in the first group by 4.7 and 11.5%, respectively. PMID- 11033863 TI - [Dependence of the kinetics of forming phases of a two-phase polymeric system on actin state]. AB - New analytical possibilities of biological macromolecule partition in a two-phase polymer system (dextran-500/polyethyleneglycol-6000) were investigated. The technical principles were based on the fact that optical density of the system changes during phase formation. Various actin forms were investigated. Unlike G actin, F-actin increases the speed of phase formation. Different actin isoforms exert different influence on this process. The kinetics of the partition system into phases makes it possible to figure out the picture of native G-actin damage. The two-phase polymer system has been shown to be capable of supporting the native properties of actin for not less than 6 days at 20 degrees C. An assumption is made that two-phase systems can be used for imitating the intracellular medium in studies on actin functional characteristics. PMID- 11033864 TI - [Effect of prenatal emotional-pain stress on the status of interphase chromatin in neurons of the rat developing brain with various excitation of the nervous system]. AB - A short-term emotional-painful stress, experienced by pregnant rat females differing in threshold of excitability of their nervous system, was used to assess the state of interphase condensed chromatin and C-heterochromatin of neuron nuclei in developing brains of 16-17 day old embryos. To reveal relationships between the genetically determined excitability of rats and the state of interphase chromatin in their neuron nuclei a computer information system has been used that enabled us to classify the neuronal nuclei according to their specific DNA image cytometry features. The results indicate an obvious relationship between excitability of the nervous system and structural-functional state of the neuronal interphase chromatin. PMID- 11033865 TI - [Interaction of superhelical DNA with the nonhistone protein HMG1]. AB - The interaction between nonhistone chromosomal protein HMG1 and plasmid DNA was studied by optical and hydrodynamical methods. The recombinant protein HMG1 produced by yeast Pichia pastoris strain was used. We have shown that according to the CD spectra local conformational changes in DNA helix occur in the region of DNA-protein interaction. These changes are most significant at r < 3 (w/w). Both gel-shift assay and ultracentrifugation, as well as CD data, indicate that protein-protein interactions between HMG1 molecules play a major role in the formation of DNA-protein complexes. It is suggested that the protein C-terminus may affect HMG1-DNA binding not only by a direct interaction with DNA helix, but also by protein-protein interactions. PMID- 11033866 TI - [Effect of mycoplasma contamination of a human cervical carcinoma cell line M HeLa clone 11 on karyotypic variability]. AB - The karyotypic variability has been investigated for an immortalized human epithelioid cervix carcinoma cell line M HeLa clone 11, cultivated for 15-60 days after contamination with Acholeplasma laidlawii A, strain PG-8, and for 30 days after contamination with Mycoplasma arginini R-16. The character of cell distribution for chromosome number changes in contaminated cells significantly, as compared to the control. So, the frequency of cells with the modal number of chromosomes being equal to 50 decreases significantly, and the range of variability in the number of chromosomes increases. With the prolongation of the term of cultivation in control variants up to 60 days the character of cell distribution for chromosomal number changes, as compared to the preceding terms (15 and 30 days), which is expressed in the extended range of variability in the chromosomal number at the expense of decreased frequency of cells with submodal number of chromosomes equal to 49. But the degree of these changes is significantly smaller than in contaminated variants. The frequency of polyploid cells did not differ in all investigated variants. The number of chromosomal aberrations in cultures contaminated with A. laidlawii (for 15-60 days) and M. arginini (for 30 days) does not differ from that in the corresponding controls. The absence of dicentrics (telomeric association) at a long-term contamination of the human epithelioid cervix carcinoma cell line M HeLa clone 11 having marker chromosomes in karyotype and a comparison of these results with the earlier obtained data on other "marker" and "markerless" cell lines seems to confirm the point of view that dicentrics appear a characteristic feature of karyotypic variability of "markerless" cell lines, mainly with a long-term contamination in different conditions. PMID- 11033867 TI - [Stereospecific anomalies in the structure of DNA from chromosomes V, XII, XIII, XIV, XV and XVI of saccharomycetes yeasts]. AB - Using an original computer program we analysed complete nucleotide sequences of chromosomes V, XII, XIII, XIV, XV, and XVI in yeast. The full catalogue of 5 highest stereospecific anomalies and of stereospecific anomalies for 5 genes with highest CAI in each chromosome has been presented. Trains of different stereospecific anomalies, possibly showing areas of cooperative binding of different regulatory and structural proteins to DNA, are documented (see: Soidla, Lukina, 1998, 1999). Together with confirming the earlier noticed association between stereospecific anomalies and genes with high expression level (coding mostly proteins of translation machinery of the cell and glycolytic enzymes), here we can also notice an obviously high incidence of transcription apparatus genes being located at (or near) largest stereospecific anomalies. Intracellular transport genes and genes whose products function in the cell nucleus may also be often associated with largest stereospecific anomalies. PMID- 11033868 TI - [Cytogenetic effects of separate and combined effect of thorium-232 and potassium nitrates on Tradescantia (clone 02)]. AB - Under both separate and combined influence of 232nd-nitrate and potassium nitrate a study was made of the yield of somatic mutation frequencies, the level of morphologically abnormal cells and the loss of reproduction integrity in the stamen hairs of Tradescantia (clone 02). All investigated concentrations of 232nd nitrate and potassium nitrate exerted statistically significant genotoxic effects and increased the level of morphological cell abnormalities in the stamen hairs of Tradescantia. Also, 0.36 mg/l 232nd and 0.51 mg/l KNO3 concentrations cause a statistically significant loss of reproduction integrity in the stamen hair. The combined action of 232nd-nitrate and potassium nitrate in all investigated concentrations and parameters causes a statistically significant effect. PMID- 11033869 TI - Feline leukemia virus and feline immunodeficiency virus. AB - Ophthalmic manifestations of FeLV or FIV infection can occur in all ocular tissues and may be manifestations of direct viral effects or secondary to viral related malignant transformation. Additionally, the manifestations of common feline ophthalmic pathogens may be more severe and poorly responsive to therapy because of the immunosuppressive effects of FeLV or FIV infection. Prompt diagnosis of underlying viral infection in cats with ophthalmic disease is paramount for accurate diagnosis and prognosis and is required for appropriate therapeutic decision making. PMID- 11033870 TI - Feline infectious peritonitis. AB - Feline infectious peritonitis is a noncurable viral disease affecting cats worldwide. Recent evidence suggests that the FIPV has evolved as a deletion mutation of FECV. Immune complex deposition and vasculitis with pyogranulomatous lesions are the hallmark of FIP. The only definitive antemortem diagnostic test for FIP is histopathologic examination of tissue. Ocular manifestations occur commonly with noneffusive FIP. The most common clinical sign is a bilateral granulomatous anterior uveitis often accompanied by chorioretinitis. Treatment of ocular FIP is symptomatic, and the mainstay of palliative therapy is topical or systemic corticosteroids or both. PMID- 11033871 TI - Feline herpesvirus. AB - Feline herpesvirus infection is extremely common and may lead to recurring ocular disease in the adult cat. Recognition of the history and clinical signs that are consistent with FHV-1 infection is critical because diagnostic tests may be negative. Although a variety of treatment options are available, no one therapy is successful in every cat, and a small percentage of cats respond poorly to any treatment. PMID- 11033872 TI - Feline chlamydia and calicivirus infections. AB - Feline conjunctivitis is common and often presents a clinical challenge to the veterinarian. Chlamydia psittaci is an important pathogen and should always be considered when evaluating cats with conjunctivitis. FCV is an infrequent cause of conjunctivitis and only causes the disease in conjunction with other clinical signs of this infection, such as oral mucosal ulcers and upper respiratory tract disease. PMID- 11033873 TI - Feline systemic fungal infections. AB - Systemic fungal diseases are important diagnostic considerations in all sick cats, particularly in cats with ocular symptoms. The most common ocular manifestation of these diseases is posterior uveitis (choroiditis); however, anterior uveitis is sometimes present and is usually secondary to the inflammation in the posterior segment. Occasionally, adnexal diseases such as blepharitis, inflammation of the nictitating membrane, and ocular discharge may be present in cats with systemic mycoses. The prognosis for cats with systemic fungal diseases has changed with the advent of the triazole antifungal drugs. In the past, the prognosis was guarded to poor for survival of the cat. Today, with prolonged antifungal therapy, many cats recover completely from their disease. The prognosis for return of vision for eyes affected with systemic fungal disease is still guarded. Often, even if the infection is controlled systemically, the retina is severely damaged and may remain nonfunctional. PMID- 11033874 TI - Toxoplasmosis. AB - Toxoplasmosis in dogs and cats can cause chorioretinitis, anterior uveitis, or both. Ocular lesions are a common manifestation of generalized toxoplasmosis. The prevalence of toxoplasmosis as a cause of idiopathic anterior uveitis in cats is not clear, although there is a significant association between exposure to T. gondii and feline anterior uveitis. The pathogenesis of ocular toxoplasmosis may be different in humans and cats, and the anterior uveitis may represent a type of immune-mediated inflammation. A diagnosis is made by observing compatible clinical findings and obtaining supportive findings on serologic tests. Despite improved diagnostic techniques, including determination of IgM class antibodies and PCR testing, definitive diagnosis of ocular toxoplasmosis remains a challenge. Topical anti-inflammatory therapy should be used in cats with anterior uveitis, a positive serum titer, and no concurrent systemic signs. Systemic clindamycin should be given to cats with ocular and systemic signs and to cats with suggestive serology and idiopathic anterior uveitis that fails to respond to topical therapy alone. PMID- 11033875 TI - Canine systemic fungal infections. AB - The principal route of infection for the disseminated fungal diseases discussed in this article is inhalation. In some cases, direct wound contamination and ingestion may also have an important role in the pathogenesis of the disease, especially in histoplasmosis. Another common theme of these diseases is the response of the immune system. If the inoculum is small and the animal is not immunocompromised, the infection may be limited to the respiratory tract and may resolve with few or no clinical signs. Dogs are usually presented to the veterinarian when the fungus has disseminated throughout the body via the circulatory or lymphatic systems, thus causing clinical signs secondary to specific organ infection. Draining skin tracts and lymphadenopathy occur in several of the diseases. The ocular location that is frequently affected is the choroid, where the organisms cause cell-mediated chorioretinitis. Early detection of these changes is important for saving vision and for diagnosing the systemic nature of the disease. Treatment is often effective, especially early in the disease, although it is expensive and long-term, with many animals needing over a year of treatment. Sometimes the treatment must continue lifelong. Ocular disease may not respond to treatment even when respiratory and other organ system clinical signs are rapidly improving. This isolation of the eye is similar to that of the CNS and requires regular monitoring of ocular disease, especially in the fundus, to ensure that systemic drugs are penetrating into the eye. Once the disease progresses to the anterior segment, the ocular prognosis worsens. Better penetration of the blood-retinal and blood-aqueous barriers may be achieved with fluconazole when compared with the other antifungal drugs. Secondary inflammatory ocular disease must also be monitored and treated appropriately to prevent scarring, which may cause vision loss or glaucoma. PMID- 11033876 TI - Canine protothecosis. AB - Canine protothecosis remains a difficult condition to manage. The paucity of clinical cases hinders the development of successful treatment strategies. The clinical signs associated with the disease are nonspecific, and the course is so insidious that, by the time a definitive diagnosis is reached, the organism has often disseminated throughout the body. At this point, the condition is beyond treatment, and death occurs owing to failure of any number of organ systems, including the gastrointestinal, cardiovascular, renal, and central nervous systems. It is of some encouragement that the few patients that have undergone aggressive early treatment have survived longer than patients presenting late in the disease course. Nevertheless, the outlook for any dog with protothecosis is grave, and it remains to be determined whether early diagnosis can truly provide a better long-term prognosis. By including protothecosis as a consideration for dogs initially brought in with a history of chronic diarrhea or acute blindness and with a subsequent finding of exudative retinal separation, early diagnosis is possible. This recognition potentially affords the opportunity for an immune status work-up and intervention with increasingly better treatment options. PMID- 11033877 TI - Canine systemic bacterial infections. AB - Although systemic bacterial diseases are an uncommon cause of uveitis in dogs, they should be included in the differential diagnoses for uveitis. A work-up for uveitis should include tests for B. canis and B. burgdorferi. If an aqueous centesis is performed, Leptospira titers or PCR can be performed on the aqueous humor and the serum. Better documentation of the role of Leptospira and B. burgdorferi in uveitis in the dog is needed. Any suspected cases should be worked up thoroughly. If the dog does prove to be positive for the organism, the case should be submitted to a peer-reviewed journal. PMID- 11033878 TI - Canine viral infections. AB - The ophthalmic effects of viral infection are varied. With the added possibility for pathologic effects of attenuated vaccine viruses, the diagnosis of viral diseases can be a challenge. In many cases, ocular manifestations can provide added support to a presumptive diagnosis of viral disease, thereby underscoring the benefit of thorough ophthalmic examination of any animal with nonspecific signs of illness. PMID- 11033879 TI - Canine rickettsial infections. AB - Dogs that live in tick-infested areas are at risk for contracting rickettsial infections. Clinical signs associated with ehrlichiosis or Rocky Mountain spotted fever may be dramatic or mild. Clinicians must consider the possibility of rickettsial diseases to request laboratory tests that will permit a proper diagnosis. Specific antimicrobial therapy usually brings about clinical improvement, although some dogs may not be cleared of rickettsial organisms, even with prolonged treatment. A small percentage of dogs die of rickettsial infections, either in the acute stage or owing to chronic bone marrow suppression and generalized debilitation. Ocular lesions are an important clinical sign in canine rickettsial infections and may aid the clinician in making a diagnosis and monitoring response to therapy. PMID- 11033880 TI - Canine and feline primary ocular bacterial infections. AB - Bacterial infections are frequently associated with diseases of the eyelids, cornea, and conjunctiva. Animals sustaining KCS commonly have bacterial infections of the external eye owing to a lack of antimicrobial properties present in the normal tearfilm. Infection can occur in the nasolacrimal duct or lacrimal sac, which is referred to as dacryocystitis. Severe corneal ulcers are frequently infected with bacteria, especially Pseudomonas sp. Three new topical ophthalmic antibiotics have recently become commercially available: ciprofloxacin, norfloxacin, and ofloxacin. PMID- 11033881 TI - [Realities and hopes of gene therapy]. AB - The work represents an introduction article of editors of special issue of the magazine devoted to gene therapy and therapeutics. The main results of clinical gene therapy in the past decade are critically considered in connection with a changes of paradigms of the field. They are: 1) change of the main target of genetic therapy--correction of defects in chromosomes--onto expression and/or output of target genes for gene therapy; 2) transfer from gene transplantation to cell transplantation; 3) tendency for the use of safe/non-viral vectors instead of viral ones.; and 4) conflict of interests in gene therapy. Outlooks in the field are discussed. PMID- 11033882 TI - [Retroviral vectors in gene therapy]. AB - This review describes the systems of retroviral transfer and expression of the genes that are widely applied in basic biological research and in gene therapy. Unique features of retroviruses providing a background for construction of retroviral vectors and the methods to use these vectors are discussed. PMID- 11033883 TI - [Nonviral gene transfer in vivo in gene therapy]. AB - Recent findings connected with in vivo use of artificial macromolecular complexes (genosomes) for functional gene transfer and delivery are discussed in the paper. Non-viral methods are the most safe for the purpose of human gene delivery. The cationic liposomes containing cholesterol are the most suitable for this purpose, because they possess high biodegradability and stability in blood stream. The DNA liposome complexes should: (i) contain DNA in the condition at most protected from environmental influence, (ii) be rather homogeneous and of small size (40-80 nm). Injections of complexes into blood are the most effective; in a respect of organospecifity may be achieved by appropriate ligand selection. It is the most perspectively to increase the expression level by combining liposomes with viral peptides. PMID- 11033884 TI - [Target delivery of functional genes in gene therapy using carbohydrate containing vectors]. AB - Various aspects of use of specific interactions with a participation of carbohydrate and oligosaccharide ligands to increase an efficiency of gene transfer into eukaryotic cells (including in vivo experiments) are considered in details. Data on addressed gene delivery with applying carbohydrate-containing ligands (such as asialoglycoproteins and galactosides) are discussed in the paper. Results on the usage of glycoside ligands, containing lactose, mannose, glucose residues, for receptor-mediated gene transfer, are analysed. Special attention is paid to application of chitosans for functional gene transfer into eukaryotic cells, which is considered by authors as a case of receptor-mediated gene transfer. It is notice that neo-oligosaccharide vectors, recognizing surface lectins, represent very perspective type of gene delivery systems. PMID- 11033885 TI - [Listeria monocytogenes: a dangerous pathogen used as a vector for the new generation of vaccines]. AB - Listeria monocytogenes (LM) has become a major pathogen of human foodborne illnesses eliciting meningitis, peritonitis, and abortions with a mortality rate of about 30%. During the course of the disease, LM infects a variety of tissues and cell types due to its capacity to induce its own phagocytosis even into non phagocytic cells. For over 35 years LM continues to serve as a model to define general paradigms of immunology In this review we focus on the clinical characteristics of listeriosis, on the risk factors involved in the pathogenesis, and discuss the currently accepted approaches to prophylaxis and treatment. We report on novel strategies in vaccine development based on the LM-dependent delivering machinery for immune recognition and induction of immunological memory against desired antigens. PMID- 11033886 TI - [Gene therapy of hereditary diseases]. AB - In the review the main advantages in development of the approaches to gene therapy of hereditary diseases are presented. Now more than 1000 genes of hereditary diseases are mapped and some hundreds are cloned which is prerequisite for gene therapy. The transfer of the recombinant gene into the cell and the subsequent expression of the transgene product are the rate-limiting steps for successful gene therapy. A variety of methods, including the use of physical methods, modified viruses and synthetic vectors, are currently being used in experiments and clinical trials. Since the approval and initiation of the first human gene therapy trial to treat ADA deficiency, there have been several dozen approved gene therapy trials but clear clinical result was stated for ADA deficiency only. Cystic Fibrosis, CF was among several hereditary diseases which were considered as a target for gene therapy. Experiments on development of recombinant gene constructions, gene delivery by adenovirus vectors and liposomes as well as by other constructions into epithelial lung cells, gene expression and on the safety of gene therapy procedures were relatively successful. Phase 1 gene therapy clinical trials of CF showed that some unaccounted physiological peculiarities of lung tissue of the patients diminished effectiveness of gene transfer, longevity of CFTR gene expression and in some cases unexpected immunological complications arises during clinical trials. Now an intensive attempt to overcome these problems in gene therapy of CF are undertaken. PMID- 11033887 TI - [Gene therapy of monogenic hereditary diseases. Duchenne myodystrophy]. AB - The paper highlights the new trends in gene therapy research area and clinical trials. It should be noted that the majority of firms involved in development of the scientific approaches to gene therapy are concentrated in the United States. The investments of the given companies on development and research of new genetic constructs also delivery systems make hundred millions dollars. The greatest part (more than 80%) of gene therapy clinical trials projects are also connected with the US research departments; the majority of them is related to tumor therapy. The advantages and drawbacks of the main methods of nucleic acids delivery to the cells are considered; diseases that are attempted to be treated using gene therapy methods are listed. A special attention of the review is devoted to the modern stand in research on cell and Duchenne muscular dystrophy (DMD) gene therapy, also brief description of basic results achieved in the authors laboratory is given. Basic original results of transfection of mdx mice (DMD biological models) with dystrophin cDNA delivered by gene gun, cationic liposomes, synthetic microspheres, viral olygopeptides and lactoferrine are summarized. PMID- 11033888 TI - [Prospects for gene therapy of cardiovascular diseases]. AB - The therapeutic potential of gene therapy in cardiovascular disease such as post angioplasty restenosis, myocardial ischaemia and severe peripheral artery disease ischemia are considered. PMID- 11033889 TI - [Gene and gene cellular therapy and neurodegenerative diseases]. AB - The modern methods of the treatment of the neurodegenerative diseases are considered. Neurodegenerative diseases originate due to the degeneration of the neuronal cells of central nervous system that leads to imbalance of the neurotransmitter synthesis and, as a consequence, movement disorders and mental disabilities. Traditional methods of pharmacotherapy and neurosurgery give short term effect. Since 1980 neurotransplantation was developed as a new technology for the treatment of the neurodegenerative diseases. This approach represents a case of cell therapy being used for transplantation of the human fetal material. Cell transplantation compensates the local deficiency of the neurotransmitter level by substitution of degenerated neurons of patient's brain (e.g. dopaminergic neurons). Gene-cell conjunction of cell therapy with modification of genome of transplanted cells is the most perspective approach to increase an efficiency of neurotransplantation. Short description of gene therapy approaches and a search for optimal gene-cell protocols for therapy of neurodegenerative diseases are presented in this paper. PMID- 11033890 TI - [Gene therapy of chronic infections of the urogenital system using cytotoxic peptides]. AB - Gene therapy of chronic infectious diseases of urogenital tract represents a new perspective field in the modern biological and medical sciences. In the review discuss one of the new directions in gene therapy of urogenital infections caused by Mycoplasma: inhibition of mycoplasmal infection after administration of recombinant plasmid vectors, expressed the genes of cytotoxic peptides. PMID- 11033891 TI - [Treatment of lumbar nerve root syndrome]. PMID- 11033892 TI - [Evidence-based medicine]. PMID- 11033893 TI - [Orthopedics, orthopedic surgery, trauma surgery--the circle is closing?!]. PMID- 11033894 TI - [The patient as expert--introduction to evidence-based orthopedics]. AB - The surgical act is based on the laws of causality from Newton and Galilei and it is determined by the principle of cause and effect. Therefore, the healing process must be interpreted as a turning back of pathogenesis or the linear chain of casualities. Scientific knowledge of this century demonstrates that biological healing processes are connected with the laws of cybernetics and the principles of semiotics. There are functional relations between the level of the organ (impairments) of the individual (disabilities) and of the society (handicaps). This International Classification of Impairments, Disabilities and Handicaps (ICIDH) serves as the key for the management of chronic diseases. An independent and a separate classification of the severity in each level is necessary to identify the consequences of the disease to the patient. Surgical interventions occur on the organ level, benefit and evidence are reflected especially on the individual level (gain of abilities). The assessment and the integration of the so-called "sensory impairment", influenced by the biographical events of the patient and the evaluation of the psychosocial resistance are important factors to recognize unfavourable conformity between the degree of impairment and the degree of disability. With this classification, a more patient-oriented discussion of the indications regarding operative procedures can be realised. The MARA model (mean age-related ability) serves as a pragmatic basis for the description of the benefits of carried out and omitted interventions as changes of abilities by using the MARA curve as an ethical guideline. This model, which is on ICIDH, the hierarchy of needs and the salutogenesis (semiotics, cybernetics), facilitate the introduction of evidence-based surgery. It helps to estimate the several predictive values and correlation factors influencing the manifestation of the disease. In this way astonishing results in evidence can be expected. Finally, many misunderstandings in health care discussions are explained by the fact that the differences between pathology and illness are not clearly interpreted. PMID- 11033895 TI - [I. Cultural adaptation, practicability and reliability evaluation of the Musculoskeletal Functional Assessment Questionnaire]. AB - INTRODUCTION: The patient-based evaluation of outcome is gaining increased importance. The aim of the study was to translate and culturally adapt the short musculoskeletal function assessment questionnaire (SMFA) into German and to evaluate the practicability, metric properties, and distribution characteristics of the German version (SMFA-D). METHODS: The SMFA is a questionnaire on the functional status of the musculoskeletal system of patients consisting of a function index (34 items) and bother index (12 items) with a 5-point response format. The SMFA was independently translated by orthopaedic-trained, qualified German- and English-speaking translators. A consensus committee review and selection of an optimally comprehensible German version was performed. Patients checked this version for comprehensibility and time to complete the questionnaire. 40 fluently German-speaking patients with primary osteoarthritis of the knee completed the SMFA-D twice at a one week interval. The test-retest reliability and internal consistency were evaluated. The values are standardized on an scale from 0-100. High values indicate a poor function. RESULTS: The SMFA-D is comprehensible and can be completed within 20 minutes in the mean. There were no skewness and no floor or ceiling effects. The mean function index was 47 (SD 17, range 16-80). The mean bother index was 44 (SD 19, range 5-75). Test-retest reliability were 0.88 (function index) and 0.71 (bother index), respectively. Internal consistency (Cronbach's alpha) was 0.92 (function index) and 0.88 (bother index), respectively. CONCLUSIONS: The SMFA-D is a practical and reliable questionnaire for patient-based evaluation of functional problems of the musculoskeletal system. PMID- 11033896 TI - [II. Validity and sensitivity to change of the Musculoskeletal Functional Assessment Questionnaire in primary gonarthrosis and total endoprosthetic joint replacement]. AB - INTRODUCTION: Scoring systems used so far in total knee arthroplasty are limited by their non-fulfillment of basic test criteria. The aim of the study was to demonstrate the validity and responsiveness of a German version of the new short musculoskeletal function assessment questionnaire (SMFA-D) in patients with primary osteoarthritis and total knee arthroplasty. METHODS: 66 patients with a tricompartmental cemented PFC-Sigma total knee arthroplasty completed the SMFA-D and the WOMAC questionnaire preoperatively and at 12 to 16 weeks follow-up. Preoperatively, physicians' rating of function of the leg, patients' self selected walking speed, pain and arthritis severity score were registered for demonstration of criterion validity. Construct validity was evaluated with the WOMAC. Discriminant validity was assessed by comparing patients with or without previous surgery at the knee, use of pain medication and use of walking aids. The function and bother indexes of the SMFA-D were correlated with these parameters. Standardized response means were calculated. RESULTS: The function index correlated with physicians' rating (r = 0.51), walking speed (r = 0.61), pain (r = 0.36) and the arthritis severity index (r = 0.36). The bother index correlated with pain (r = 0.37) and the arthritis severity index (r = 0.25). The function and bother index correlated with the WOMAC (r = 0.77) and (r = 0.81), respectively. Patient groups with or without walking aids (p = 0.02) and with or without pain medication (p = 0.001) differed in the function index. The bother index of patients with or without pain medication (p < 0.001) and with or without walking aids (p < 0.006) differed. Function and bother index improved from 46 (SD 17) to 34 (SD 19, p < 0.001) and 43 (SD 18) to 33 (SD 22 p < 0.001), respectively. The standardized response mean was 0.86 for the function index and 0.53 for the bother index. CONCLUSIONS: The SMFA-D questionnaire is valid and responsive in patients with primary osteoarthritis of the knee and total knee arthroplasty. It measures function and bother from the perspective of these patients. PMID- 11033897 TI - [Results of cement-free St. Nabor hip prosthesis implantation and clinical and radiological results after 8 to 10 years]. AB - PURPOSE OF THE STUDY: The aim of the retrospective study was to analyse the medium-term results of the cementless hip prosthesis St. Nabor. METHOD: 46 patients with complete postoperative and radiographic documentation were available for clinical review. The average follow-up was eight years and five months (range from eight years, one month to ten years). The average age at the last follow-up was 60 years. Ten patients showed severe coxarthritis and 32 patients a dysplasia of the acetabulum before surgery. Two patients suffered from a severe osteonecrosis of the femoral head and two patients had fractures of the acetabulum or femoral neck, respectively. RESULTS: The medium-term follow-up for the acetabular component showed a survival rate of 97.8%. The radiographic data visualized one loosening as a result of a low grade infection, 18 months after surgery. There was no migration of the acetabular component according to Sutherland et al. and no protrusion was found. None of the acetabular components showed radiolucent lines or was loose according to radiographic criteria. In comparison to the first post-operative radiograph, 26 of the hips were found with a significant reduction of lacunae, twelve of them showed a complete absence at last follow-up. The overall Harris hip score averaged 92 points (range 58 to 100 points) for the entire group. 41 (92%) of all hips were rated as excellent or good. Two hips (4%) of all hips were rated as fair and two hips (4%) were rated as poor. CONCLUSION: The high rate of good and satisfying results in medium-term follow-up proves this St. Nabor acetabular component system to be an excellent implant in total hip arthroplasty for younger patients with or without dysplasia. PMID- 11033898 TI - [Mannheim concept of preoperative autologous blood donation and perioperative autologous blood transfusion in hip endoprosthesis surgery]. AB - PURPOSE: To investigate the efficiency of preoperative autologous deposit and intra- and postoperative cell salvage (CS) to reduce homologous transfusion in hip arthroplasty and revision hip arthroplasty. METHODS: In a retrospective study, the data of 1402 patients scheduled for hip arthroplasty and for revision hip arthroplasty were analysed. RESULTS: 767 women and 635 men, age 62.9 +/- 9.8 years (x +/- s) were included in the study. 1270 were scheduled for hip arthroplasty, 132 for revision hip arthroplasty. Of the autologous donors, 51 patients predeposited four units, 1020 patients three, 204 patients two, 39 patients one unit. 88 patients who had not enrolled in the autologous donation program but received CS served as a control group. Blood loss in autologous donors amounted to 1620 (220-5620) ml in hip arthroplasty and 2830 (950-7910) ml in revision arthroplasty. CS was employed in part of the cases in arthroplasty and in all revision operations. 470 (0-2200) ml and 705 (0-2200) were retransfused. In hip arthroplasty 227 of 1182 patients (19.2%) received homologous blood. Homologous transfusion in patients with coxarthrosis due to acetabular protrusio, avascular necrosis of the femoral head and hip dysplasia showed a trend to higher values. Patients who had donated 3 units and received CS required homologous transfusion in 12.4% of the cases. CS reduced the homolgous transfusion rate significantly if the preoperative hemoglobin concentration was < or = 12 g/dl. A group of autologous donors receiving CS was matched with patients receiving CS only. 12 of 76 (15.8%) vs. 45 of 76 (59.2%) required homologous transfusion. In revision hip arthroplasty 58 of 132 patients (43.9%) required homologous blood. CONCLUSIONS: Preoperative deposit reduces homologous transfusion requirements considerably in hip surgery. Under the conditions studied CS should be employed in hip arthroplasty in addition to preoperative deposit if the preoperative hemoglobin concentration falls below 12 g/dl. In revision arthroplasty, 4 or more autologous units should be predeposited and CS should be used regularly. PMID- 11033899 TI - [Endoprosthetic management of patients with hip arthrodesis]. AB - PURPOSE: Purpose of this retrospective study was to investigate the long-term results of total hip replacement in patients with hip arthrodesis. METHODS: Between 1976 and 1986 conversion of a earlier performed arthrodesis of the hip to total hip replacement was carried out in a total of 20 patients. 15 of these have been followed up postoperatively for an average of 15.7 years. The main preoperative symptoms were pain affecting the lumbar spine, the ipsilateral knee and/or both hip joints. In two cases, the arthrodesis has fractured. RESULTS: With the exception of one female patient, all the patients expressed satisfaction with the postoperative results, pain having been alleviated or eliminated. The difference in leg length presenting preoperatively in 12 patients was partly or completely equalized in ten cases. Specific physiotherapy over a period of several months resulted in an appreciable increase in muscular power in all patients. Postoperatively ten patients had a slight, five a pronounced limp, while 11 patients needed a walking aid. Recurrent dislocations occurred in one patient. We observed one aseptic loosening of the cup after 10 years. CONCLUSION: The results of the study show that conversion of surgically ankylosed hip joints to total hip replacement makes good sense. Provided that the indication is established with care, this major surgical procedure represents a suitable method, not only of improving the patient's quality of life, but also, and in particular, of at least appreciably improving the symptoms developing after hip arthrodesis. There seems no increased rate of aseptic implant loosening. PMID- 11033900 TI - [Hip joint operation in routine management--complications and their effect on indications-specific symptoms]. AB - AIM: The study was conducted in co-operation with a German sickness fund to identify determinants of disease-specific health outcomes after hip surgery in routine health care. METHOD: In September 1997 all beneficiaries (age 40-75 yrs.), who were hospitalized for "osteoarthrosis" (ICD 9-715/820), were sent a disease-specific survey instrument on average 5.2 months (T1) after discharge. Survey content focused, among others, on pre- and postoperative symptoms and impairment (Lequesne index), postoperative complications, comorbidity (Katz Index) and health-related quality of life (SF-36). The response rate at T1 was 67.8%. Only patients undergoing hip surgery were sent a second survey instrument 17.2 months (T2) after discharge. After two mailings, data from 293 patients were available for analysis. Descriptive and multivariate analysis (GSK Model) were performed. RESULTS: Patients (57.6% male) were on average 61 yrs. of age and 61.2% reported no comorbidity. 88.4% received total hip replacement. A third of patients reported at least one complication. Univariate, a substantial (and highly significant) improvement was found for the Lequesne Index over time: (recalled) preoperative: 14.2 points; postoperative T1: 5.6 pts.; postoperative T2: 4.4 pts. This result is confirmed by multivariate analyses (estimated values: pre = 13.8, T1 = 6.9; T2 = 5.7), although it is modified by an interaction effect between the variables "Lequesne index" and "complication". The Lequesne Index in patients with complications is estimated preoperative 12.9, at T1 = 7.3 and T2 = 6.2. The respective estimated values for patients without complications are: preoperative 14.7, T1 = 6.5, T2 = 5.2. CONCLUSION: Patients with postoperative complications obtain lower scores on the Lequesne Index (higher burden of disease) before hip surgery. Their postoperative progress is slower in the short term (-5.6 pts. vs. -8.2 pts) and in the medium term (-6.7 pts. vs. -9.5 pts). Complications after hip surgery have a lasting negative effect on disease specific health outcomes. PMID- 11033901 TI - [Significance of subsequent hip unloading after surgical intertrochanteric osteotomy of idiopathic femur head necrosis]. AB - PURPOSE: To judge the influence of consequent non-weight-bearing on the postoperative results, we reviewed 28 patients in whom we performed intertrochanteric osteotomies for idiopathic osteonecrosis of the femoral head during the years 1996 and 1997. Postoperatively 12 patients were supplied with a walking device for non-weight-bearing which was used for 1 year. 16 patients were mobilized by using 2 crutches for 15 kp partial weight-bearing until the osteotomy healed up and were then allowed to bear full weight. METHOD: For clinical evaluation of the early results after about 18 month the Merle d'Aubigne hip score was used. Furthermore we analyzed pelvic radiographs for assessing the stage of the necrosis (Ficat staging) and the necrotic angle. RESULTS: Overall 82% good and excellent results were achieved. Due to a longer period of non weight-bearing the results were again clearly improved (92% versus 75% of patients with partial weight-bearing). It was noticed that there were discrepancies between good clinical results and the less satisfying radiographic findings. CONCLUSIONS: Postoperative non-weight-bearing for about 1 year with the use of a walking device by patients with idiopathic osteonecrosis of the femoral head has clearly a positive effect on clinical results. PMID- 11033902 TI - [Significance of calcium deposit elimination in tendinosis calcarea of the shoulder]. AB - AIM: To evaluate the effect of complete elimination of the calcific deposit on the clinical outcome in patients with a recalcitrant calcifying tendinitis of the shoulder. PATIENTS AND METHODS: 50 patients with a chronic calcifying tendinitis of the shoulder (Type Gartner I or II) underwent a single application of extracorporeal shock waves (3000 impulses, energy flux density 0.60 mJ/mm2). After one year 45 patients were evaluated radiologically and clinically. RESULTS: Radiologically a complete absorption of the calcific deposit was observed in 21 patients (47%). In 9 patients there was no radiological change. The radiological success was closely related to the radiomorphology of the deposit. 18 out of 27 good or excellent results were accompanied by a complete disintegration of the deposit. We calculated a significant dependency between complete disintegration of the deposit and a good/excellent clinical result (p = 0.0002). Patients with a completely disintegrated deposit had significantly better scores than those with a partial disintegration of the calcium (p = 0.02), and than those with no radiomorphological changes (p = 0.0003). Spearman's correlation coefficient was 0.6. CONCLUSION: Elimination of the deposit is paramount for a good clinical outcome and should be the main goal for any treatment. PMID- 11033903 TI - [Traumatic rupture of the rotator cuff in an adolescent--case report]. AB - A traumatic rotator cuff tear is a very rare condition, especially in the adolescent. We report about a 14-year-old boy who fell onto the right arm while riding his bicycle. Immediately after the accident he described some kind of click in his right shoulder. he was referred to our institution 3 days after the trauma. The MRI showed a huge defect of the labrumligament complex including the rotator interval. Intraoperatively, we found a complete disruption of the subscapularis and supraspinatus tendons as well as of the anterior stabilizers. We performed a modified Bankart repair and a reconstruction of the ruptured rotator cuff. The patient returned to his former sports as cyclist 3 months after surgery. This case is of clinical relevance because of the traumatic disruption of an intact rotator cuff in a young patient. Not the bone but rather the tendons and ligaments were the weakest link in this situation. In cases of primary traumatic shoulder dislocation, we have to look for bony and soft-tissue damage including the rotator cuff. With the present high standards in ultrasound and MR imaging we have to keep these techniques in mind in order not to overlook this type of accompanying injury. PMID- 11033904 TI - [3-dimensional image analysis of the shoulder joint--a new method for characterizing parameters of shoulder joint function]. AB - OBJECTIVE: An exact assessment of shoulder movement is of special importance both in the diagnosis of and in the therapy for different shoulder diseases. Therefore, we developed a feasible method for the analysis of shoulder movement. METHODS: On the basis of an ELITE system and 6 skin markers (marker positions. acromion, humerus, olecranon, proc. styloideus ulnae, cervical and thoracic spine), movement analysis was performed during continuous abduction of the arm over 15 s. With the help of a purpose made software we determined the exact angle of abduction and, in addition, the acceleration (+aAC) and the deceleration ( aAC) of the acromion. We evaluated 12 normal subjects (10 male, 2 female, average age 29 yrs) without shoulder conditions and 8 patients (6 male, 2 female, average age 46 yrs.) with unilateral impingement syndrome stage II according to Neer. RESULTS: In addition to a significantly diminished abduction ability in patients with impingement syndrome, our results also revealed significantly decreased acceleration values for the acromion in impingement patients. In contrast, deceleration values for the acromion were not altered in patients with impingement syndrome. CONCLUSION: The presented method allows exact measurements of shoulder movement. In addition, measurements of acromion acceleration and deceleration seem to offer two parameters for the assessment of shoulder function in pathological conditions. Further investigations are required to prove the advantages and limitations of this method. PMID- 11033905 TI - [Surgical peri- and postoperative plexus anesthesia in subacromial decompression of the shoulder]. AB - AIM: The aim of the present investigation was to check the peri- and postoperative plexus anesthesia on early functional outcome and postoperative analgetic consumption during subacromialen decompression until the date of discharge. METHODS: 50 patients in the period 01.01.1997 to 01.10.1998 treated by an arthroscopic or open subacromial decompression of the shoulder were registered. Pre- and postoperative functional movement and postoperative analgetic consumption within the first 12 days after the operation were compared according to the procedure used. RESULTS: The peri- and postoperative plexus anesthesia had a positive influence to early functional outcome as well as on the analgetic consumption. The results are shown in the group of arthroscopic (18) surgery and in the group of the patients operated in an open (32) technique. The patients with plexus anesthesia showed a better functional movement of the shoulder directly postoperatively and a lower consumption analgetic drags. At the time of discharge we found no significant differences between the subjects in both groups. CONCLUSION: The investigation shows that peri- and postoperative plexus anesthesia has no positive influence on the early functional outcome after subacromial decompression of the shoulder. PMID- 11033906 TI - [Scanning stereographic surface measurement in idiopathic scoliosis after VDS (ventral derotation spondylodesis)]. AB - INTRODUCTION: So far only radiometric and clinical methods have been available for the evaluation of results after anterior scoliosis surgery. Rasterstereography has proved to be a reliable method for three-dimensional surface measurement of conservatively treated idiopathic scoliosis patients. Therefore, patients treated operatively with anterior instrumentation were examined using rasterstereography to determine the three-dimensional correction of the spinal deformity. The aim was to measure back shape deformity, in particular derotation, and thus cosmetic improvements. METHODS: 31 patients with idiopathic thoracic, thoracolumbar and lumbar scoliosis (Cobb angle 57.2 degrees) were examined with raster stereography preoperatively, postoperatively and after follow-up (25.2 months) in a standardized standing posture. Standing radiographs were compared with raster stereography. RESULTS: The mean Cobb angle was reduced from 57.2 degrees to 17.2 degrees, the rasterstereographic maximal surface rotation from 16.5 degrees to 10.8 degrees, and the vertebral rotation according to Perdriolle from 29.2 degrees to 16.7 degrees. During follow-up the Cobb angle increased to 20.8 degrees, and surface rotation to 11.3 degrees. Vertebral rotation remained constant. Lordosis and kyphosis angles changed only slightly. CONCLUSION: Rasterstereography is a suitable tool for analyzing the three dimensional correction of spinal deformities after anterior scoliosis surgery. In particular, the cosmetic improvement is clearly demonstrated. The measurement of surface rotation allows objective quantification of the obtained derotation. PMID- 11033907 TI - [Early clinical screening in psoriatic arthropathy]. AB - AIM: Because of doubts in early clinical assessment and full documentation of all various cases of psoriatic arthritis outside of rheumatologic centres a clinical study in a dermatologic department of a rehabilitation clinic was performed. METHODS: 148 patients between 1996 and 1999 were interviewed and examined clinically. 27 (18.2%) of them had osseous joint processes proved by X-ray (severe cases). 90 (60.8%) patients had no radiologic changes but a positive history of joint pain (mild cases). 31 (21%) patients had none of the problems (Control group). Blood specimens were taken in order to reveal inflammations (ESR, CRP, RF, ASL, uric acid, and leucocytes). Exclusion criteria were other arthritides, multiple osteoarthritis and consolidated fractures of joints. The groups were compared together. RESULTS: Whereas palpable joint crepitations did not turn out as a reliable specific sign, pressure pain on acromioclavicular joints and MTP joints, impingement signs of the shoulders, deformities of toes and flexor tendon swellings of the hand were usable tools in diagnosing a severe psoriatic arthritis in this study. The ESR, ASL, CRP, and uric acid were elevated significantly more often in severe than in mild cases. Moreover, the time period between the first onset of skin lesions and joint pain was shorter in severe cases than in mild cases (9.86 vs 13.8 years). CONCLUSION: The diagnostic relevance of criteria, for example of the well-known European Spondylarthropathy Group (ESSG), may be improved by new early screening criteria. PMID- 11033908 TI - [Psychogenic joint contractures exemplified by somatoform hysteria]. AB - We would like to report about a 25-year-old female patient with a psychogenic knee contracture as part of a hysterical personality disorder. After having had multiple surgical procedures including 5 knee arthroscopies in 5 different orthopedic departments, we found a nearly full range of movement during an examination under general anaesthesia. Psychoanalytical investigations finally revealed a surprising, past history of the patient and her family. In addition we highlight the relevant literature. PMID- 11033910 TI - Current concepts in diagnosis and treatment of posterior cruciate ligament injury. AB - Injuries to the posterior cruciate ligament (PCL) have traditionally been reported to be rare but in the last decade, this ligament has captured the attention of orthopaedic surgeons world-wide. While new or improved surgical techniques involving the PCL have achieved attractive results, accurate evaluation of the PCL-deficient knee still remains a challenge. Despite the greater use of new, more sensitive clinical testing, stress radiography and MRI, correct diagnosis and staging of the PCL tear is still difficult. The objective of this paper is to provide an up-to-date review regarding diagnosis and treatment of the PCL tear. PMID- 11033909 TI - [Comment on "Warning about carbon fiber pins"]. PMID- 11033911 TI - [The syndrome of external ligament sprain in the horse]. AB - The treatment of acute complete (grade III) tears of the lateral ligaments of the ankle has generated much controversy in the medical literature. Functional treatment has become the standard treatment as it has been shown that there is no significant difference in long term results whatever the treatment (operative repair and cast, cast alone, or early controlled mobilization). Functional treatment includes only a short period of protection by tape bandage or brace and allows early weight-bearing. Major trauma with avulsion of bone and severe ligamentous damage on both medial and lateral sides of the ankle is however an indication for surgical treatment in the acute phase. Secondary operative reconstruction can be performed in case of persistent instability and laxity of the ankle. Secondary anatomic repair as proposed by Brodstrom, Duquennoy et al. and Karlsson et al. has a high rate of success and avoids the potential morbidity of harvesting partially or totally the peroneus brevis or other tendon grafts. Evaluation of the injured ankle has improved and in selected patients ultrasonography, arthrography, magnetic resonance imaging or bone scintigraphy may be useful for further evaluation of the injury. The frequency of associated injuries has probably been underestimated. Although ankle sprain is often thought of as an injury involving only the lateral ankle ligaments, there are varied and multiple components to the common sprained ankle. This condition would perhaps more appropriately be designated as the sprained ankle syndrome. PMID- 11033912 TI - Spinal instrumentation for primary pyogenic infection report of 31 patients. AB - The role of spinal instrumentation in the presence of infection is still controversial. Radical debridements of infected vertebrae and disc material and bone grafting usually leaves the spine unstable without some surgical stabilisation. We reviewed 31 cases of primary pyogenic spinal infection treated by radical debridement, bone grafting and posterior (30) or anterior (1) spinal instrumentation. The indication for surgery was the failure of conservative treatment (8), progressive neurological deficit (19) or the lack of diagnosis (3). The clinical, laboratory and radiological parameters were assessed pre and postoperatively. The mean period of follow-up was 3.8 years (1-12 years). The neurological deficit was progressive in 19 patients, following surgery all these patients were improved. The neurological deficit was established in one patient; following surgery, his neurological deficit did not improve. The infection was eradicated in all our patients. The following complications were encountered: (1) three patients developed deep wound infection, which responded to repeated debridement; (2) one death resulted from nosocomial septicaemia, (3) reoperation was carried out on one patient for implant failure and on another for a dislodged anterior bone graft. We conclude that spinal instrumentation may be indicated when after radical debridement of infected vertebrae and disc material and bone grafting the stability of the spine is still compromised. According to the location of the infection and the availability of suitable implants, anterior or posterior instrumentation may be necessary. With appropriate antimicrobial agents, the outcome has been satisfactory in our patients. PMID- 11033913 TI - The Boytchev procedure for recurrent anterior dislocation of the shoulder. A controversial technique. AB - In the Boytchev technique the conjoined tendons of the coracobrachialis and short head of the biceps, together with the tendon of the pectoralis minor and the detached tip of the coracoid process are rerouted behind the subscapularis muscle, and reattached to the coracoid process with a screw. During a 10-year period 37 patients with recurrent anterior dislocation underwent a Boytchev procedure. Twenty-six patients (with 27 shoulders) were available to follow-up. The results of this retrospective study were disappointing, as the overall redislocation rate was 44%. The opinions of other authors were found to be extremely divergent. PMID- 11033914 TI - The Sauve-Kapandji procedure for posttraumatic wrist disorders: further experience. AB - A prospective survey was conducted to evaluate the outcome of the Sauve-Kapandji procedure for posttraumatic wrist disorders. Eighty four patients were treated, all with posttraumatic disorders of the distal radioulnar joint, 73 as an isolated procedure, 11 in combination with another wrist procedure. There was significant pain decrease and high patient satisfaction (74%). The range of motion increased in the flexion/extension arc from 109 degrees to 124 degrees (p = 0.006) and, in those with limited forearm rotation, from 71 degrees to 134 degrees (p = 0.006). According to the Mayo Clinic wrist score, we obtained 20 excellent, 34 good, 18 fair and 12 poor results. Complications were rare. PMID- 11033915 TI - Ethnic factors in Perthes disease: a retrospective study among white and Asian population living in the same environment. AB - The aim of this study was to find out about the incidence of Perthes disease in different ethnic groups living in similar environments. Among the child population of Bradford, between 1991-1997, we identified from the file notes twenty-five cases of Perthes disease among the white European population (with an incidence of 5:54500 for the year 1997) and one case of Perthes disease among Asians (with an incidence of 0:16100 for the year 1997). There were two positive family histories of Perthes disease and only a case of bilateral Perthes disease. There was no significant difference in the incidence of Perthes disease among the rural and urban population. We conclude that under similar environmental circumstances the genetic factor plays an important role in the predisposition to Perthes disease. PMID- 11033916 TI - Assessment of hip rotation after gait improvement surgery in cerebral palsy. AB - Sixteen children with diplegic type of cerebral palsy and spastic internal rotation gait were evaluated using gait analysis before and an average of 3 years after multiple soft tissue surgery. Significant correction of the internal rotation gait was observed after multi-level soft tissue surgery which included medial hamstring lengthening in all cases. Our results suggest that for children with spastic internal rotation gait, multi-level soft tissue surgery effectively corrects the dynamic internal rotation gait in the absence of fixed bony rotational deformities. PMID- 11033917 TI - One-stage bilateral total hip arthroplasty a simultaneous procedure in 79 patients. AB - The authors retrospectively studied 79 patients who had undergone simultaneous bilateral total hip arthroplasty during the period 1982 to 1994. Forty one patients were examined clinically and radiographically at least 5 years postsurgery. The procedure was associated with few early postoperative complications and so far excellent results at 7.5 years with regard to patient satisfaction, Hip Functional Index and survival of the prostheses. It is concluded, that in selected patients with bilateral hip disease necessitating bilateral hip replacement, the bilateral operation may be advantageously carried out in one session. PMID- 11033918 TI - Treatment of osteoarthritis of the knee with valgus deformity by means of varus osteotomy. AB - Nineteen cases of osteoarthritis of the knee with valgus deformity in 17 patients were treated by femoral supracondylar varus osteotomy (17 cases) or by high tibial varus osteotomy (2 cases) over the last 15 years. Fixation was performed using a 95 degrees AO blade-plate in 13 of the femoral osteotomies and a straight plate in the other four. The mean follow-up time was 6.5 years. The valgus deformity was idiopathic in 14 cases, secondary to rheumatoid arthritis in 2 cases and to tibial valgus in one case. The Hospital for Special Surgery (HSS) score was used to evaluate the clinical results: nearly 75% were excellent or good. The causes related to poor results are analyzed regarding indication and surgical technique. It appears that varus osteotomy is an effective procedure for the treatment of osteoarthritis of the knee with valgus deformity, above all in order to alleviate pain, although the operation requires precision, and correct selection of patients is of prime importance. PMID- 11033919 TI - Ilizarov treatment of tibial nonunions results in 16 cases. AB - Treatment with the Ilizarov technique was performed in 16 patients with complex tibial nonunions. Two years post treatment the functional stage and patient satisfaction were recorded. There were 4 hypertrophic, 3 atrophic and 9 infected nonunions. Eleven patients had segmental bone loss. Fifteen nonunions united, and limb length discrepancy was reduced within 1.5 cm of the contralateral leg. Average time in the frame was 182 days. Fifteen of the 16 patients were satisfied with the treatment. One patient demanded an amputation after 3 months of treatment, despite good signs of healing. There were no refractures or recurrent infections. In conclusion the Ilizarov technique for complex nonunions has a high rate of success in achieving union and eradicating infection, bone loss and malalignment. The treatment is demanding both to the surgeon and to the patient, but we strongly recommend the Ilizarov treatment for tibial nonunion, especially in cases with chronic infection and severe bone loss. PMID- 11033920 TI - Chondrosarcoma of the distal phalanx of a toe. A case report. AB - Secondary malignant transformation of a solitary enchondroma into a chondrosarcoma is extremely rare. We report a case of such transformation in anenchondroma of the distal phalanx of a toe. PMID- 11033921 TI - Metallosis mimicking osteomyelitis from a forearm plate retained for 50 years. AB - A forearm plate retained for 50 years was removed because an acute inflammation mimicked chronic infection. The radiographs were very confusing. During the operation metallosis was obvious. Despite the general opinion that forearm plates should be kept in place, this probably should not apply to old "historical" hardware of uncertain chemical composition. PMID- 11033922 TI - Accidental subcutaneous injection of elemental mercury. A case report. AB - Subcutaneous injection of elemental mercury is a very rare situation. The authors report the case of a 31-year-old man who accidentally injected an unknown quantity of metallic mercury into his left forearm. Several surgical procedures were required to reduce the blood and urinary levels of mercury. However, the patient never developed clinical signs of chronic poisoning. This observation confirms the lower risk of acute or chronic poisoning in subcutaneous injection of mercury and the need for early excision of contaminated tissue. PMID- 11033923 TI - Stress fracture of the inferior and superior pubic ramus in a man with anorexia nervosa and hypogonadism. AB - The authors present the case of a 23-year-old athlete with a bifocal stress fracture of the pubis. Bifocal cases are rather rare, therefore an insufficiency fracture was suspected. Further investigation showed osteopenia due to secondary hypogonadotropic hypogonadism caused by anorexia nervosa. This case illustrates the need for awareness of underlying metabolic disorders in very rare cases of stress fractures in athletes. PMID- 11033924 TI - Realignment of tibial deformity in Paget's disease. A case report. AB - Paget's disease of bone is a relatively common condition which usually is asymptomatic. Furthermore, there is limited experience in surgical procedures, especially in realignment of severe skeletal deformities. Likewise, there are often relapses in tibial deformities treated with plates. We report a case of a severe tibial deformity treated with multiple osteotomies and stabilized with an unreamed medullary nail. PMID- 11033925 TI - Bilateral fractured clavicles--a pair of cases. AB - The authors present two cases of bilateral fractured clavicles, both resulting from direct trauma to both shoulders. In both cases, the fractures healed uneventfully with non-operative treatment. PMID- 11033926 TI - Grandparents as parents: research findings and policy recommendations. AB - This article presents an overview of research on grandparenthood in the latter decades of the twentieth century. Theories contributing to understanding of the grandparenting role are discussed, and significant factors affecting the grandparenting experience--including sex, age, retirement status, race, and ethnicity--are reviewed. The special case of grandparents raising grandchildren is explored through a review of demographics, outcomes for children in grandparent foster care, and the impact of raising grandchildren on grandparents. Interventions supporting custodial grandparents and the grandchildren in their care are examined. Drawing on the findings and implications of this overview, recommendations for policy, clinical practice, professional education, and future research are offered. PMID- 11033927 TI - Interparental conflict and adolescent depressed mood: the role of family functioning. AB - Family functioning was investigated as a mediator between interparental conflict and adolescent depressed mood among adolescents living in two-parent and divorced families. Data were collected three times, with one year intervals. At the initial interview, adolescents were, on the average, 15.5 years old. Two types of interparental conflict were assessed: interparental conflict involving arguments about the adolescent, and arguments focused on the parents' behaviors. The results confirmed that family functioning mediated the effects of parent issue/interparental conflict, but not adolescent issue/interparental conflict. This was most evident for girls than boys. Implications of the findings for research and practice are discussed. PMID- 11033928 TI - Perceptions of control in children with externalizing and mixed behavior disorders. AB - Examined the differences in the perception of control in 67 school-age children with externalizing and children with both externalizing and internalizing behavior problems. The results indicated that children with externalizing behavior and mixed behavior could be differentiated by their perception of control. Specifically, children with externalizing behavior endorsed a significantly stronger unknown locus of control than children in the mixed behavior group. Findings suggest that when behavior groups are clearly defined, neither external nor internal locus of control is the dominant style. Implications for the findings are discussed. PMID- 11033929 TI - A parental report on the long-term consequences for children of abduction by the other parent. AB - Little is known about the long-term impact on the child of abduction by a parent. Children who had been kidnapped by one of their parents and hidden for an average of 2.7 years have been followed for a decade through contact with the parent who recovered them. This study reports on the most recent series of interviews. It was hypothesized that children who were now late teens and young adults would, on the whole, be progressing satisfactorily into young adulthood and that relationships with their parents would be non-problematic. According to 32 parental reports gleaned both from a telephone interview with open-ended and closed-ended questions and from a brief mailed questionnaire with closed-ended questions, a significant minority of the children continue to suffer emotionally and may be having more physical ailments than their peers. Relations with recovering parents were non-problematic. PMID- 11033930 TI - Parental assessment of behavior in Chinese adoptees during early childhood. AB - The Parent Rating Scale of the Behavior Assessment Scale for Children (PRS-BASC) was used to examine the behavior of 45 Chinese adoptees. In all but one case, results from the 9 BASC-PRS scales ranged in the average, or normal, range. Thus, as a group, no deviations from normal behavior were revealed. However, the variability of ratings for several scales--Hyperactivity, Aggression, Conduct Problems, and Attention Problems--indicated a greater potential for at-risk behavior. Age of adoption from China was not a significant influence on parents' perceptions of adoptees' behavior. However, older adoptees were more likely to be rated hyperactive or aggressive than younger children, while younger adoptees were more likely to exhibit withdrawal. PMID- 11033931 TI - Men in nursing: the importance of gender diversity. PMID- 11033932 TI - Education. A nurse is a nurse is.... PMID- 11033933 TI - International affairs. Reflections. PMID- 11033934 TI - Legal and ethical issues. Ethical considerations in international nursing. PMID- 11033935 TI - Problem-based learning for nursing: integrating lessons from other disciplines with nursing experiences. AB - Problem-based learning (PBL) is a philosophy and teaching method that structures knowledge in clinical contexts, strengthens motivation to learn, develops clinical reasoning skills, and enhances self-directed and life-long learning. During the past 30 years, PBL has been implemented in schools of medicine worldwide, including 100 of the 126 schools in the United States. This article reviews the international PBL literature and focuses on the key aspects of this paradigm in educating health professionals: recent history and worldwide diffusion, basic characteristics and rationale, typical case presentation, student and tutor roles, and data-based outcomes. Conclusions from selected meta analytic studies of assessment are described and discussed. The potential of PBL for nursing education is examined. Key implementation issues are identified, including the challenge to persuade faculty to move from efficient teaching to effective learning. PMID- 11033936 TI - Legal aspects of grading and student progression. AB - In today's litigious environment, nurse educators must understand the legal issues involved in determining grades and deciding academic progression. Although the courts have consistently ruled that educators are the most qualified people to evaluate what constitutes academic success, the potential remains for students to legally challenge their grades. This article reviews legal cases and shows the change in the student-institution relationship from one of a child-surrogate parent (in loco parentis) to one in which students' rights and institutional responsibility are balanced. The courts continue to support the concept that educators are uniquely qualified to make determinations concerning grading and progression and defer to decisions made by the academic community. However, a series of legal decisions based on due process and contract law now protects students from arbitrary and capricious decisions made without a formal grievance process. The implications for nurse educators and administrators are far-reaching and include such issues as specific policies within syllabi, divisional policies, institutional policies, and adherence to formal grievance procedures. PMID- 11033937 TI - Implementing Web-based instruction in a school of nursing: implications for faculty and students. AB - The World Wide Web, a rich educational resource that can promote and enhance student learning, is increasingly being used in nursing programs to deliver course content. When implementing Web-based instruction into the curriculum, it is essential to consider carefully the implications for faculty and student development needs so that the technology can be efficiently and effectively used to support student learning. Teaching a Web-based course is a new experience for many faculty and requires a reconceptualization of the faculty role. Faculty issues to consider when implementing Web-based instruction include instructional design concerns, faculty-student interactions, time and technology management skills, and student outcome evaluation. Students, especially those who have a preference for faculty-directed classroom learning, also will find student role challenges in Web-based learning. Time and technology management skills, student faculty interactions, and becoming more self-directed in their learning are student role development needs. This article describes the strategies used by one school of nursing to meet faculty and student development needs when the RN-BSN completion program was redesigned for Web-based instruction. PMID- 11033938 TI - Characteristics of nursing doctoral programs in the United States. AB - The expansive growth of doctoral programs in the United States has necessitated a need to determine demographic attributes of the academic program, characteristics of the students, admission criteria, and curriculum patterns. Seventy-eight doctoral programs were sent a researcher-designed survey and 48 (62 per cent) responded. Results show an increasing shift away from clinical doctorates to the research doctorate, although few differences exist within the research focus of doctoral nursing programs. Consistent with previous reports in the literature, curricula are fairly standard and few differences were noted based on a number of criteria including geographic location, type of degree granted, Carnegie classification of the university, or by the length of time the program has been in existence. Discussion of these findings is included. PMID- 11033939 TI - Development and implementation of a semester program in Mexico for senior nursing students. AB - The College of Nursing (CON), Michigan State University (MSU), in collaboration with the School of Nursing and Obstetrics, University of Guanajuato, Celaya, Mexico, developed a semester-long study-abroad program for senior MSU nursing students offered for the first time in the fall of 1998. The program provides intensive Spanish language classes and allows students to take required nursing courses in Mexico with a substantial amount of course content provided by Mexican faculty without an MSU CON faculty member on site at all times. Students receive a broad perspective of nursing and health care in Mexico, and develop an appreciation for its language and culture as well. This program represents an innovative approach to the development and implementation of a study-abroad program in nursing. PMID- 11033940 TI - Emphasis courses: preparing baccalaureate students for transition to the workforce. AB - In response to identified needs for stronger preparation of baccalaureate graduates to enter the workforce, the University of Maryland School of Nursing created, implemented, and evaluated the use of Clinical Emphasis Seminars and Practicum. A sequence of 3 courses was ultimately developed for students to take during the final year of their upper-division nursing curriculum. The courses, totaling 5 credits, allow students to develop their knowledge and experience in a focus area within 1 specific type of clinical nursing. These courses follow up completion of required foundational clinical courses. Emphasis areas include all clinical areas offered within the nursing curriculum such as cardiovascular, critical care, and community health. Courses begin and end with 1-credit seminars and include one 3-credit clinical Practicum. In the latter course, students work with a clinical preceptor, following the preceptor's work hours for a minimum of 9 hours a week over the 15-week semester. Advantages of the Emphasis Courses are many: students develop an in-depth knowledge and clinical skills in a selected area of clinical practice. Potential employers have an opportunity to observe students in a preemployment setting and to evaluate the fit of skills and personal attributes to the specific setting. Students are provided with "value added" knowledge and experience. The emphasis course model offers advantages to students transitioning into the workforce, to employers, and, ultimately, to clients served. PMID- 11033941 TI - [Diagnostic and surgical treatment of chronic abdominal aneurysm rupture]. AB - Examination and treatment of 8 patients with the abdominal aorta aneurysm (AAA) chronic rupture (CHR) was done. The interval between the aneurysm rupture occurrence and of the operation performance had constituted 3.6 months. The disease was diagnosed basing on the anamnesis data, on the results of clinical, invasive and noninvasive methods of investigation. The authors suggest necessary to operate all the patients with the AAA CHR. PMID- 11033942 TI - [The volume and consistency of surgical intervention for the affected several extracranial vessels]. AB - The results of the cerebral hemodynamics investigation and of operative treatment of 94 patients with affection of several extracranial vessels were presented. It was established that along with the total arterial lumen narrowing stage increase the pronounced cerebral tissue hypoperfusion occurs. Therefore the reconstruction of several arteries is necessary. In patients with low cerebral perfusional reserve it is expedient to proceed with revascularization in stages. PMID- 11033943 TI - [Contemporary approach to the treatment of complicated diabetic angiopathy of the lower extremities]. AB - The experience of treatment of 278 patients with diabetes mellitus, complicated by the lower extremities diabetic angiopathy, was summarized. The authors had proposed the complex of treatment, including combined application of the high- and low-energy laser irradiation and catheterization of the shin arteries branches and the anterior abdominal wall superficial arteries for an adequate introduction of medicinal agents towards the affection focus. Application of the method proposed had promoted the results of treatment improvement, had permitted to proceed with economical amputation of extremity and to escape the reoperations performance. PMID- 11033944 TI - [Persisting viral infection, serum antibodies and chromosome instability in myasthenia]. AB - There were examined 94 patients, aged from 22 to 68 years, with myasthenia, in whom morphological changes of thymic gland were revealed. In 89% of observations there was established the infectioning by cytomegalovirus, in 10%--hepatitis B, in 9%--hepatitis C. In 5% of patients there were observed coexistence of cytomegaloviral infection and hepatitis. In all the patients the trustworthy increase of the chromosomal aberrations frequency in comparison with spontaneous level of mutation was revealed. High diagnostic validity of chain reaction with polymerase in the viruses persistence determination was noted. The results of investigation of serous immunoglobulins to the virus neutralizing antibodies ratio may be applied for the indications elaboration in the lymphosorption and immunocorrection application in patients with myasthenia more than 4 years in course. PMID- 11033945 TI - [The pleural effusion syndrome in patient with tumor of thymus]. AB - Five observations of the thymic gland tumor and concurrent exudate pleuritis were presented. In one patient the pleuritis was total and in the others--restricted one, in two of the patients exudate was diagnosed during performance of operation. In two observations in exudate there were revealed tumoral cells. In two patients tumor turned out to be inoperable one, the rest of the patients were operated on successfully. PMID- 11033946 TI - [The state of regional hemodynamics after performance of operations of the portogastroesophageal blood flow disconnection in liver cirrhosis]. AB - There were examined 112 patients, to whom the various kinds of separating operations were done. After the gastroesophageal junction suturing performance the even venous pressure raising in all parts of the stomach was noted. After performance of the original intervention, directed on the hemorrhage from varicose gastroesophageal veins elimination, the pressure in the high gastric portion had lowered and hyperdynamical state of blood circulation in this zone had normalized. The operation performance for separation of gastroesophageal venous system from the v. porta system had promoted the pressure lowering in upper portion of stomach veins and in v. gastroepiploica. After performance of the operations, directed on separation of portoesophageal blood flow, the pressure and volumetric speed of the blood flow in v. porta had raised. PMID- 11033947 TI - [Laparoscopic interventions on choledochus: 5 year experience]. AB - Complex examination of 69 patients with complicated cholelithic disease was performed. The elaborated in the clinic algorithm for preoperative examination of the patients was applied to reveal the main bile ducts pathology. Indications for the laparoscopic interventions on choledochus performance were substantiated. PMID- 11033948 TI - [Optimization of complex treatment of purulent cholangitis]. AB - The results of treatment of 53 patients with purulent cholangitis were analyzed. In complex of treatment there were applied hemocarboperfusion and nondirect electrochemical oxygenation of the blood. It had promoted the lowering of the endogenic intoxication severity and the oxygen-transporting system of blood improvement. PMID- 11033949 TI - [Surgical treatment of patients with damage and cicatricial stricture of biliary ducts]. AB - In 109 patients 106 reconstructive and 3 restorational operations were performed for iatrogenic damage and cicatricial structure of biliary ducts. Formation of biliobiliary anastomosis is effective in the treatment of the stricture restricted in length. The preference was given for hepaticoduodenostomy performance on the changeable transhepatic drain (in 60.4% of observations) or for the elaborated in the clinic method of operation application when high stricture was present or when the broad anastomosis formation was impossible. Application of optimal algorithm for surgical tactics had permitted to lower the mortality rate for 5-year period from 8.2 to 6.8%. PMID- 11033950 TI - [Considering advantages of dynamical omentopancreatostomy in the treatment of necrotic pancreatitis]. AB - While application of dynamical omentopancreatopexy for the treatment of necrotic pancreatitis there were noted the purulent and visceral complications frequency lowering from 3.34 in every patient--while the closed interventions usage down to 2.02 and of mortality--from 57.2 down to 33% accordingly. PMID- 11033951 TI - [The restoration of intestinal continuity after the performance of the Miculicz's operation]. AB - The method of double-barrelled (split) colostomy closure after performance of Miculicz operation, including extraperitoneal formation of the invagination duplicatural anastomosis, was proposed. After application of the proposed method in 21 patients the complications occurred in 2, including 1 case of the anastomotic sutures insufficiency. PMID- 11033952 TI - [Surgical strategies in concurrent closed thoracoabdominal trauma]. AB - The experience of surgical treatment of 393 injured persons with closed thoracoabdominal trauma (CTAT) in an acute period was summarized. The CTAT structure, peculiarities of diagnosis, surgical tactics, complications were regarded and the injured persons deaths causes were analyzed. Operative intervention was performed in 202 (51.4%) injured persons. Mortality was 35.1%. PMID- 11033953 TI - [Relaparotomy: clinical, strategic and organizational aspects]. AB - The results of relaparotomy (RL) performance in 118 patients were analyzed. Mortality was 16.1%. In 50.8% of patients RL was done with delay. There were proposed for timely diagnosis of postoperative complications to use the "alarm" symptoms group and during RL performance--to follow definite technical methods and rules. The RL performance necessity in 62.7% patients was caused by surgical technique failures and nonrational method of the first operation choice. PMID- 11033954 TI - [Efficiency of surgical treatment of patients with pulmonary tuberculosis and diabetes mellitus in conditions of the mycobacterial tuberculosis resistant to pharmacotherapy]. AB - The peculiarities of surgical treatment of 32 patients with destructive pulmonary tuberculosis and diabetes mellitus, in whom the Mycobacterium tuberculosis resistance to chemopreparations was revealed, were enlighted. The measures of optimal preoperative preparation and postoperative patients management were substantiated. Postoperative complications occurred in 62.5% of patients. Clinical effect was achieved in 78.12% of patients. PMID- 11033955 TI - [Differences in differential diagnosis between acute mesenterial lymphadenitis and appendicitis in children]. AB - The thermal streams intensity (THSI) from definite points of anterior abdominal wall in children with an acute mesenterial lymphadenitis (AML) and an acute appendicitis (AA) were determined. It was established, that in right inguinal region THSI in AA was in 4.06 times higher than in AML, in paraumbilical region- THSI was in 2.78 times, in the left subcostal region--in 1.76 times higher in AML than in AA. In the right subcostal region THSI was higher in an AA. Differential algorithm for AML and AA was determined. In children with an AML the circulating immune complexes level in the blood plasma was by 26.4% higher than such in children with an AA, presenting possible additional differential-diagnostical criterion. PMID- 11033956 TI - [Operative treatment of subcutaneous rupture of achilles tendon]. AB - The indications for the tendon suture and autoplasty application for subcutaneous tendo calcaneus (Achilles) rupture were established as well as possibilities of the local anesthesy application during performance of operative intervention. The enzymotherapy performance after the operation had permitted to reduce the complications frequency. PMID- 11033957 TI - [Detoxication therapy in children: individualized approach to the application of the hemodiafiltration method]. AB - The toxicity investigation technology was elaborated, permitting to individualize the indications for application of the hemodiafiltration method in children. There was proposed the method of the blood serum separation on the compounding fractions with subsequent determination of cytolytical activity (toxicity) of every substance with respect to proper leukocytes. PMID- 11033958 TI - [Conversion in laparoscopic cholecystectomy: failure, timely strategies and decisions or pre-planned change of strategies]. PMID- 11033959 TI - [The role of a students' club in the formation of the future surgeon]. PMID- 11033960 TI - [Pathogenesis of acute biliary pancreatitis]. PMID- 11033961 TI - [Lymphogenic complications in renal transplantation]. PMID- 11033962 TI - [The late diagnosis and surgical treatment of the cardiac and pericardium injuries]. PMID- 11033963 TI - [Sigmoid colon perforation during colonoscopy]. PMID- 11033964 TI - [Formation of interintestinal anastomosis with temporary two-barrel intestinal stoma]. PMID- 11033965 TI - [The performance of aortocoronary shunting in a child aged 9 month]. PMID- 11033966 TI - [The mitral valve prosthesis combined with "labyrinth" approach method]. PMID- 11033967 TI - [Congenital fistula between the coronary and pulmonary arteries]. PMID- 11033968 TI - [The application of video-assisted thoracoscopy in diagnosis and treatment of the thoracic cavity organ diseases]. PMID- 11033969 TI - [Combined thoracoabdominal injury]. PMID- 11033970 TI - [The penetrating injury of thorax and neck]. PMID- 11033971 TI - [The diagnostic experience of the small intestine tumors]. PMID- 11033972 TI - [Coexistence of perforative duodenal ulcer and incarcerated umbilical hernia]. PMID- 11033973 TI - [Traumatic rupture of two spleens and a liver in a child]. PMID- 11033974 TI - [Observation of successful treatment of granulomatous hepatitis in a child]. PMID- 11033975 TI - Glycemic status and soluble tumor necrosis factor receptor levels in relation to plasma leptin concentrations among normal weight and overweight US men. AB - OBJECTIVE: Leptin, an adipocyte-derived protein product of the obesity (ob) gene, is a multifunctional polypeptide associated with the development of obesity related disorders in humans. There is considerable inter-individual variation in plasma leptin even among subjects with comparable obesity levels, which suggests that factors other than adipose mass may be involved in the regulation of leptin expression and/or production. The purpose of this study was to evaluate the potential role of glycemic status and adipose-derived cytokines in regulating plasma leptin levels among normal and overweight men. DESIGN: Cross-sectional study. SUBJECTS AND MEASUREMENTS: We measured plasma leptin, insulin, c-peptide and plasma soluble tumor necrosis factor receptor (sTNF-R) concentrations in 178 men. The subjects were selected from the Health Professionals Follow-up Study (HPFS), and aged 47-64 y in 1994, were free of cardiovascular disease, diabetes mellitus, malignant neoplasms, and had provided a fasting blood sample and a detailed lifestyle questionnaire. RESULTS: Men in the highest quintile of plasma leptin (mean = 12.7 ng/ml) weighed more, were less physically active and had higher circulating insulin, c-peptide, sTNF-R1 and sTNF-R2 concentrations than men in the lowest quintile (mean = 2.8 ng/ml). We found a significant correlation between plasma insulin, c-peptide, glycosylated hemoglobin (HbA1c), and sTNF-R1 on leptin concentrations (with Spearman correlation coefficients ranging from 0.17 to 0.48 and all P < 0.05). Only HbA1c and sTNF-R1 were independently and positively associated with plasma leptin after further adjusting for body mass index and other metabolic parameters of interest. Interestingly, these observed associations were limited to men with a BMI > or = 25 kg/m2. CONCLUSION: Our results suggest that glucose homeostasis and the activity of the TNF system may modulate leptin secretion and production among overweight men. Glucose homeostasis and TNF-alpha is important in metabolic disorders related to hyperleptinemia. PMID- 11033976 TI - Profile of body composition of school children (6-18y) in Bahrain. AB - OBJECTIVE: To study the body composition of Bahraini school children. DESIGN: Cross-sectional national school survey. SUBJECTS: School children aged 6-18y. The sample comprised 818 boys and 775 girls. MEASUREMENTS: Weight, height, arm circumference and skinfold thicknesses were measured. Sum of skinfold thickness, body mass index (BMI), mid arm circumference, percentage body fat, fat weight and lean body weight were then calculated to determine body composition. RESULTS: A significant difference in the sum of skinfold thickness was observed between boys and girls. The girls have almost 50% extra skinfold thickness than boys at all ages. Similar trends were found in BMI, except at ages 9 and 18y. The percentage of body fat was higher in girls than boys. The mean BMI for Bahraini girls aged 13y and above exceeds that of their American counterparts, indicating a trend towards fat accumulation in the Bahraini girls. CONCLUSION: The data obtained are useful for monitoring obesity in school children in Bahrain as well as being usable as reference data for similar countries in the region. The high proportion of body fat among Bahraini school children, especially girls, urges an intervention program to prevent and control obesity in this age group. PMID- 11033977 TI - Lack of association between lipaemia and central adiposity in subjects with an atherogenic lipoprotein phenotype (ALP). AB - OBJECTIVE: To investigate the associations between indices of adiposity and cardiovascular risk factors in individuals with an atherogenic lipoprotein phenotype (ALP). SUBJECTS: Fifty-five men, aged 34-69 y, body mass index (BMI) 22 35 kg/m2, with an ALP lipid profile (triglycerides (TG) 1.5-4.0 mmol/l, HDL<1.1 mmol/l; %LDL-3>40% total LDL). DESIGN: Each participant provided a fasting blood sample and underwent an 8 h postprandial assessment and had anthropometric measurements taken. OUTCOME MEASURES: BMI, waist circumference (W), waist-to-hip ratio (W/H), sum of skinfolds (SSK), fasting and postprandial concentrations of glucose, insulin and plasma lipids, post-heparin lipase activity, and apoE genotype. RESULTS: The expected positive associations between BMI, W and SSK and fasting and postprandial insulin were observed (r=0.42-0.65). Little association between glucose responses and any measures of adiposity was evident. Unexpectedly, there were no positive associations between measures of central adiposity (W and W/H) and fasting and postprandial TG responses, with a trend towards negative associations in this study group (TG AUC vs W, r=-0.23, P=0.097; TG IAUC vs W/H, r=-0.26, P=0.068). Subgroup analysis indicated that lack of a positive association between central adiposity and postprandial TG values was more evident in those with one E4 allele (r=-0.42, P=0.077) relative to non-E4 carriers (r=-0.16, P=0.430). The expected positive associations between insulin and TG responses were not observed (r=-0.03 to -0.36). CONCLUSION: In this ALP group the expected positive association between TG responses and a centralized distribution of body fat was not observed, particularly in individuals with an apoE4 genotype. Our findings are not in line with the view that there is a clear causal relationship between insulin resistance and the lipid abnormalities associated with ALP. PMID- 11033978 TI - Can anyone successfully control their weight? Findings of a three year community based study of men and women. AB - This study examined the prevalence, distribution and correlates of successful weight loss and successful weight maintenance over three years in a community based sample of 854 subjects aged 20-45 at baseline. More than half (53.7%) of the participants in the study gained weight within the first twelve months, only one in four (24.5%) successfully avoided weight gain over three years, and less than one in twenty (4.6%) lost and maintained weight successfully. The findings underscore the importance of current public health efforts to prevent weight gain, and suggest that without much greater efforts to promote and support weight control the prevalence of obesity will continue to rise. PMID- 11033979 TI - Validity of self-reported weight and height in the French GAZEL cohort. AB - OBJECTIVE: To examine the validity of self-reported weight and height and the resulting body mass index (BMI), and to explore the associations between demographic, socioeconomic, and health-related factors on the one hand and bias in self-reported weight and height on the other, in order to determine the groups most likely to exhibit bias. DESIGN: Prospective cohort study. SUBJECTS: 7350 middle-aged subjects, 5445 men and 1905 women, from the GAZEL cohort, who have been followed up since 1989 and work at the French national company Electricite De France-Gaz De France (EDF-GDF) in various occupations. MEASUREMENTS: Self reported weight and height were based on information from yearly mailed questionnaires, and measured weight and height, used here as true values, were provided by occupational physicians from 1994 to 1997. Sex, age, marital status, education, occupation, history of ischemic heart disease, and treatment for cardiovascular risk factors were obtained from the mailed questionnaires or from data provided by the Company's personnel and medical departments. RESULTS: Strong correlations were found between measured and self-reported values, but self reported weight and height displayed significant systematic errors. Weight was significantly underestimated for men (0.54 kg) and for women (0.85 kg), and height overestimated for men (0.38 cm) and women (0.40 cm). These biases led to significant underestimations of BMI (0.29 and 0.44 kg/m2 for men and women respectively). Consequently, the prevalence of overweight, defined as BMI > 26.9 kg/m2 for women and BMI > 27.2 kg/m2 for men, was also underestimated, by 13% for men and 17% for women. The five factors associated with bias in self-reported weight and height were: overweight status, end-digit preference, age, educational level and occupation. CONCLUSION: These findings suggest that self-reported weight and height should be treated with caution, because of biases leading to misclassification for overweight and obesity, especially in certain segments of the population. PMID- 11033980 TI - Critical evaluation of energy intake using the Goldberg cut-off for energy intake:basal metabolic rate. A practical guide to its calculation, use and limitations. AB - OBJECTIVES: To re-state the principles underlying the Goldberg cut-off for identifying under-reporters of energy intake, re-examine the physiological principles and update the values to be substituted into the equation for calculating the cut-off, and to examine its use and limitations. RESULTS: New values are suggested for each element of the Goldberg equation. The physical activity level (PAL) for comparison with energy intake:basal metabolic rate (EI:BMR) should be selected to reflect the population under study; the PAL value of 1.55 x BMR is not necessarily the value of choice. The suggested value for average within-subject variation in energy intake is 23% (unchanged), but other sources of variation are increased in the light of new data. For within-subject variation in measured and estimated BMR, 4% and 8.5% respectively are suggested (previously 2.5% and 8%), and for total between-subject variation in PAL, the suggested value is 15% (previously 12.5%). The effect of these changes is to widen the confidence limits and reduce the sensitivity of the cut-off. CONCLUSIONS: The Goldberg cut-off can be used to evaluate the mean population bias in reported energy intake, but information on the activity or lifestyle of the population is needed to choose a suitable PAL energy requirement for comparison. Sensitivity for identifying under-reporters at the individual level is limited. In epidemiological studies information on home, leisure and occupational activity is essential in order to assign subjects to low, medium or high PAL levels before calculating the cut-offs. In small studies, it is desirable to measure energy expenditure, or to calculate individual energy requirements, and to compare energy intake directly with energy expenditure. PMID- 11033981 TI - Inhibitory effects of chondroitin sulfate prepared from salmon nasal cartilage on fat storage in mice fed a high-fat diet. AB - OBJECTIVE: Chondroitin sulfate is an acidic polymer consisting of repeating D glucuronic acid and D-N-acetylgalactosamine units, and the N-acetylgalactosamine is substituted with the sulfate at either the 4' or 6' position, with approximately one sulfate being present per disaccharide unit. The present study assessed the effects of chondroitin sulfate on the activity of pancreatic lipase and lipid uptake into brush border membrane vesicles of the rat small intestine in vitro, and on the degree of fat storage induced in mice by the oral administration of a high-fat diet for 8 weeks. DESIGN AND MEASUREMENTS: Experiments were carried out to clarify whether or not chondroitin sulfate inhibited pancreatic lipase activity in assay systems using triolein emulsified with phosphatidylcholine or gum arabic. In addition, the effects of chondroitin sulfate on lipid absorption by brush border membrane vesicles were examined. Moreover, mice were fed a high-fat diet and treated with chondroitin sulfate for 8 weeks. RESULTS: Chondroitin sulfate dose-dependently inhibited the pancreatic lipase activity in an assay system using triolein emulsified with phosphatidylcholine. In addition, chondroitin sulfate inhibited the palmitic acid uptake into the brush border membrane vesicles of the rat jejunum. Chondroitin sulfate caused the reduction of body weight and parametrial adipose tissue weight, and prevention of fatty liver and hyperlipidemia in mice fed a high-fat diet. CONCLUSION: The reduction of fat storage and the antihyperlipidemic action of chondroitin sulfate might be due to the inhibition of small intestinal absorption of dietary fat through the inhibition of pancreatic lipase activity and fatty acid uptake through brush border membrane. PMID- 11033982 TI - Leptin plasma concentrations are dependent on body fat distribution in obese patients. AB - AIM: To evaluate whether fat distribution plays a role in determining serum leptin concentrations. PATIENTS AND METHODS: One-hundred and forty-seven obese patients, 77 males and 70 females, aged 45.1 +/- 13.2 y (mean +/- s.d.; range 21 73 y), with body mass index (BMI) ranging from 30 to 55 kg/m2 (mean 42.3 +/- 5.9). Ultrasound assessment of the thickness of subcutaneous and preperitoneal fat was carried out and calculation of their ratio as abdominal fat index (AFI), waist-hip ratio (WHR), body composition by bioelectrical impedance to evaluate the percentage of fat mass (FM%) and total amount of fat (FMKg) were also determined. Plasma leptin was measured by radio immuno assay (RIA). RESULTS: In the whole group of patients, serum leptin concentrations were 37.2 +/- 18.4 ng/ml (range 6-101.3 ng/ml); in spite of BMI values not being significantly different, women had leptin values significantly higher (47.4 +/- 17.4 ng/ml) (P < 0.01) than males (28.1 +/- 15.1 ng/ml), also after correction for fat mass. The mean thickness of abdominal subcutaneous fat was 33.7 +/- 12.9 mm and it was significantly (P < 0.001) higher in female (40.9 +/- 10.6 mm) than in male (27.1 +/- 11.2 mm) patients; preperitoneal thickness was 22.9 +/- 7.1 mm, with significantly (P < 0.05) higher values in males (24.2 +/- 6.8 mm) than in females (21.7 +/- 7.3 mm). Accordingly, AFI (in all patients 0.84 +/- 0.6) was significantly higher in males (1.09 +/- 0.6) than in females (0.56 +/- 0.2). In the overall population, leptin concentrations were directly and significantly related to subcutaneous but not preperitoneal fat; they showed a strong inverse relationship with AFI and WHR. When the results were evaluated dividing the patients according to gender, subcutaneous fat thickness showed a stronger association with leptin levels in males than in females, whereas no association was found with preperitoneal fat thickness. Leptin and AFI values were significantly related only in men. WHR values were not correlated with leptin concentrations in either sex. When fat mass was added to the model, subcutaneous fat thickness, AFI and WHR remained independently associated with leptin concentrations. Age and diabetes did not influence these measures. CONCLUSIONS: Fat distribution contributes to the variability in serum leptin in obese patients. In particular, subcutaneous abdominal fat is a determinant of leptin concentration, also independently of the amount of fat mass, whereas the contribution of preperitoneal visceral fat is not significant. PMID- 11033983 TI - Indirect estimates of body composition are useful for groups but unreliable in individuals. AB - OBJECTIVE: To assess the usefulness of the body mass index (BMI) in identifying individuals classified as overweight or obese based on estimates of body fat percentage (BF%) obtained by the deuterium dilution (BF%DD) method. In addition, to assess the accuracy of bioelectrical impedance analysis (BIA) and skinfold thickness (SFT) measurements in the estimation of body composition of Australians at the individual and group level. DESIGN: Cross-sectional study. SUBJECTS: One hundred and seventeen healthy Australian volunteers of European descent, comprising of 51 males and 66 females, ranging in age from 19 to 77 y. MEASUREMENTS: BMI was calculated from body weight and height. Fat-free mass (FFM) was estimated from measures of total body water (TBW) using deuterium dilution (FFM(DD)), SFT using the equations of Durnin and Womersley (Br J Nutr 1974; 32: 77-97) (FFM(SFT)), and BIA using the equations of Lukaski et al (J Appl Physiol 1986; 60: 1327-1332) (FFM(Lu)), Segal et al (Am J Clin Nutr 1988; 47: 7-14) (FFM(Se)) and Heitmann (Eur J Clin Nutr 1990; 44: 831-837) (FFM(He)). Estimates of fat mass (FM) were calculated as the difference between body weight and FFM, while BF% was calculated by expressing FM as a percentage of body weight. RESULTS: BMI had poor sensitivity and positive predictive value in identifying individuals as being overweight/obese as classified by BF%DD. Furthermore, estimates of FFM (and hence FM) from BIA or SFT could not be used interchangeably with DD, without the risk of considerable error at the individual level. At the group level errors were relatively smaller, though statistically significant. While FFM(SFT) could be corrected by the addition of the bias (1.2 kg in males and 0.8 kg in females), no simple correction was possible with BIA estimates of FFM for any of the equations used. However, an accurate prediction of FFM(DD) was possible from the combination of FFM(He), biceps SFT and mid-arm circumference in both males and females. The bias of this prediction was small (<0.15 kg), statistically non-significant in both sexes, and unrelated to the mean FFM obtained by the two methods. The revision of Heitmann's estimate of FFM using anthropometric variables described in this study had the best sensitivity (79%), specificity (96%) and positive predictive value (92%) in identifying overweight/obese individuals in comparison to the other equations tested. CONCLUSION: BMI was a poor surrogate for body fatness in both males and females. The currently recommended equations for the prediction of body composition from SFT and BIA provided inaccurate estimates of FFM both at the individual and group level as compared to estimates from DD. However, Heitmann's equations, when combined with measures of the biceps SFT and mid-arm circumference, provided better estimates of FFM both at the individual and group level. PMID- 11033984 TI - Body composition and resting energy expenditure in humans: role of fat, fat-free mass and extracellular fluid. AB - OBJECTIVE: The objective of this study was to determine whether there are independent effects of extracellular fluid volume (ECF) and fat mass (FM) on resting energy expenditure (REE) relative to fat-free mass (FFM) in adult men and women. METHODS: Multiple linear regression analysis was used to relate REE, as determined by indirect calorimetry, to FFM and FM (measured using dual energy X ray absorptiometry) and ECF (measured using bromide space and/or the radiosulfate washout space) in 153 women and 100 men with varying amounts of body fat. RESULTS: REE correlated significantly with FFM and FM in women (r=0.65 and r=0.63, both P<0.001) and men (r=0.62 and r=0.48, both P<0.001, FFM and FM, respectively). In a multiple linear regression analysis FFM, FM and age significantly contributed to the ability to predict REE in both genders. The models that were derived were not significantly different between women and men. In women the contribution to REE from FM was easier to detect when FM was greater. Adjustment of FFM for ECF did not improve the relationship between FFM and REE. CONCLUSIONS: FFM, FM and age are significant, independent predictors of REE in both men and women. Adjustment of FFM for ECF does not improve the ability of FFM to predict REE, which suggests that ECF is a highly integrated component of FFM in healthy adults. Expressing REE relative to FFM alone will introduce errors when lean and obese populations are compared. PMID- 11033985 TI - Endogenous fat oxidation during medium chain versus long chain triglyceride feeding in healthy women. AB - OBJECTIVE: To compare the effect of medium chain triglycerides (MCT) vs long chain triglycerides (LCT) feeding on exogenous and endogenous oxidation of long chain saturated fatty acids (LCSFA) in women. SUBJECTS: Twelve healthy female subjects (age 19-26 y, body mass index (BMI) 17.5-28.6 kg/m2) DESIGN AND MEASUREMENTS: In a randomized cross-over design, subjects were fed weight maintenance diets providing 15%, 45% and 40% of energy as protein, carbohydrate and fat, respectively, with 80% of this fat comprising either a combination of butter and coconut oil (MCT) or beef tallow (LCT). Following 6 days of feeding, subjects were given daily oral doses of 1-(13)C labelled-myristic, -palmitic and stearic acids for 8 days. Expired 13CO2 was used as an index of LCSFA oxidation with CO2 production assessed by respiratory gas exchange. RESULTS: No difference in exogenous LCSFA oxidation was observed as a function of diet on day 7. On day 14, greater combined cumulative fractional LCSFA oxidation (16.9 +/- 2.5%/5.5 h vs 9.1 +/- 1.2%/5.5 h, P < 0.007), net LCSFA oxidation (2956 +/- 413 mg/5.5 h vs 1669 +/- 224 mg/5.5 h, P < 0.01), and percentage dietary LCSFA contribution to total fat oxidation (16.3 +/- 2.3%/5.5 h vs 9.5 +/- 1.5%/5.5 h; P < 0.01) were observed in women fed the MCT vs LCT diet. With the MCT diet, but not the LCT diet, combined cumulative fractional LCSFA oxidation (P < 0.03), net LCSFA oxidation (P < 0.03), and percentage dietary LCSFA contribution to total fat oxidation (P < 0.02) were increased at day 14 as compared to day 7. Day 14 results indicated increased endogenous LCSFA oxidation during MCT feeding. CONCLUSION: The capacity of MCT to increase endogenous oxidation of LCSFA suggests a role for MCT in body weight control over the long term. PMID- 11033986 TI - Effects of food attributes on hunger and food intake. AB - OBJECTIVE: To explore the relative importance of a food's macronutrient composition, energy value, energy density, fiber content, weight, volume, sensory properties and rheology on hunger and food intake. DESIGN: Preloads of peanuts, peanut butter (rheology control), almonds (tree nut), chestnuts (macronutrient control), chocolate (sensory control), rice cakes (volume control), pickles (weight control) and no load (time control) were consumed by subjects in random order at weekly intervals and hunger was assessed over the subsequent 180 min. Free-feeding energy and macronutrient intake were monitored 24 h before and following preload ingestion. SUBJECTS: Twelve male and 12 female healthy, normal weight (12-28% body fat), adults (mean (s.d.) age 22 +/- 2.5 y) with low dietary restraint. RESULTS: Hunger ratings following consumption of the 2092 kJ (500 kcal) preloads of peanuts, peanut butter, almonds, chestnuts and chocolate were significantly lower than the low energy preloads or no preload condition, but with the exception of peanut butter, did not vary from each other. The rate of hunger recovery was consistent across all preloads so the overall impact of each food on hunger was determined by the initial drop it evoked. Total energy, but not macronutrient, compensation was observed with all preloads. Consequently, the fatty acid profile of the total diet reflected the composition of the preloads. CONCLUSIONS: Energy content may be the primary determinant of a food's impact on hunger. Because macronutrient compensation is weak, a dietary supplement or substitute may influence the daily dietary nutrient profile. PMID- 11033987 TI - A five-year trend of increasing obesity among elementary schoolchildren in multiethnic, low-income, inner-city neighborhoods in Montreal, Canada. AB - OBJECTIVE: To describe the prevalence of overweight and obesity over 5 y among inner-city elementary schoolchildren aged 10-12 y in multiethnic, low-income neighborhoods in Montreal, Canada. DESIGN: Height and weight of all students in grades 4-6 in 16 control schools participating in an evaluation of the impact of a school-based heart health promotion program, were measured each May/June from 1993 to 1997 in cross-sectional classroom-based school surveys. RESULTS: The prevalence of overweight (> or = 85th age- and gender-specific percentile for body mass index (BMI) from NHANES I and II) was 35.9% in 1993; it increased by approximately 1.3% per year. The prevalence of obesity (> or = 95th age- and gender-specific percentile for BMI from NHANES I and II) was 15.9% in 1993; it increased by approximately 1.0% per year. In multivariate models predicting overweight and obesity, the odds ratios for year adjusted for age, sex, and family ethnic origin were 1.08 (95% confidence interval (CI), 1.04-1.12), and 1.09 (95%CI, 1.04-1.15), respectively. CONCLUSION: There were significant secular trends of increasing overweight and obesity among young inner-city schoolchildren from the early to late 1990s. Preventive intervention is clearly indicated because childhood obesity tracks to adulthood and because obesity is associated with substantial morbidity, mortality and health care costs. PMID- 11033988 TI - Relation of leptin and insulin to adiposity-associated elevations in sympathetic activity with age in humans. AB - OBJECTIVE: To determine whether plasma leptin and insulin concentrations are related to adiposity-associated elevations in muscle sympathetic nerve activity (MSNA) with age in healthy adult humans. DESIGN: Cross-sectional investigation of young and older adult men. SUBJECTS: Thirty healthy adult men, 16 young (25+/-1 y, mean+/-s.e.) and 14 older (61+/-1 y). MEASUREMENTS/RESULTS: The older men had higher (P<0.05) levels of body mass, BMI, total fat mass and truncal fat mass (dual energy X-ray absorptiometry) than the young men. MSNA burst frequency (microneurography) was approximately 75% higher in the older men (P<0.001). Plasma leptin concentrations were approximately 150% higher (P<0.01), whereas plasma insulin concentrations were approximately 70% higher (P<0.05) in the older subjects. MSNA was related to both total (r=0.51, P<0.01) and truncal (r=0.56, P<0.01) fat mass. Plasma leptin concentrations were related to total and truncal fat mass (both r=0.83, P<0.001), and to MSNA (r=0.49, P<0.01). Plasma insulin concentrations were related to MSNA (r=0.38, P<0.05). We used partial correlation analyses to assess whether leptin and/or insulin are potential contributors to the relation between body fat and MSNA. Adjusting for the effects of plasma leptin, but not insulin, concentrations eliminated the significant relations between MSNA and total and truncal fat mass. CONCLUSION: Our results: (1) demonstrate a positive relation between MSNA and plasma leptin concentrations in young and older healthy men; and (2) support the concept that circulating leptin concentrations may act as a humoral signal contributing to adiposity-associated elevations in MSNA with age in adult humans. PMID- 11033989 TI - Body mass index in a US national sample of Asian Americans: effects of nativity, years since immigration and socioeconomic status. AB - OBJECTIVE: To examine body mass index (BMI) and the proportion overweight and obese among adults age 18-59 in the six largest Asian American ethnic groups (Chinese, Filipino, Asian Indian, Japanese, Korean, Vietnamese), and investigate whether BMI varies by nativity (foreign-vs native-born), years in US, or socioeconomic status. DESIGN: Cross-sectional interview data were pooled from the 1992-1995 National Health Interview Survey (NHIS). SUBJECTS: 254,153 persons aged 18-59 included in the 1992-1995 NHIS. Sample sizes range from 816 to 1940 for each of six Asian American ethnic groups. MEASUREMENTS: Self-reported height and weight used to calculate BMI and classify individuals as overweight (BMI > or = 25 kg/m2) or obese (BMI > or = 30 kg/m2), age, sex, years in the US, household income and household size. RESULTS: For men, the percentage overweight ranges from 17% of Vietnamese to 42% of Japanese, while the total male population is 57% overweight. For women, the percentage overweight ranges from 9% of Vietnamese and Chinese to 25% of Asian Indians, while the total female population is 38% overweight. The percentage of Asian Americans classified as obese is very low. Adjusted for age and ethnicity, the odds ratio for obese is 3.5 for women and 4.0 for men for US-vs foreign-born. Among the foreign-born, more years in the US is associated with higher risk of being overweight or obese. The association between household income for women is similar for US-born Asian Americans and Whites and Blacks, but is much weaker for foreign-born Asian Americans. CONCLUSIONS: While these data find low proportions of Asian Americans overweight at present, they also imply the proportion will increase with more US-born Asian Americans and longer duration in the US. PMID- 11033990 TI - Pro12Ala mutation in the peroxisome proliferator-activated receptor gamma2 (PPARgamma2) and severe obesity: a case-control study. AB - OBJECTIVE: To explore the association of the Pro12Ala mutation in the peroxisome proliferator-activated receptor gamma2 with severe obesity and the features of the metabolic syndrome in a population-based sample of Caucasians. PARTICIPANTS AND METHODS: The study is based on a case-control design: 95 non-diabetic severely obese (body mass index, BMI > 35 kg/m2) cases and 280 normal weight (BMI < 25 kg/m2), age- and sex-matched controls selected from the same population were studied. Height, weight, waist circumference, as well as blood pressure were measured according to a standard protocol. BMI at age 25 y was calculated on the basis of current height and reported weight at age 25 y Biochemical measurements included fasting glucose, triglycerides, high-density lipoprotein cholesterol and insulin. DNA analysis was conducted by PCR and gel electrophoresis. RESULTS: Age and gender distribution were similar in obese and normal weight participants. The percentage of people with the Pro12Ala mutation was not significantly different in obese or normal weight participants (20% and 15%, respectively; P = 0.32). Conversely, in obese participants with obesity starting in early adulthood (ie with BMI at age 25 above 26.9kg/m2 which represents the median of the whole obese group), the Pro12Ala mutation was observed significantly more frequently than in the normal weight controls (29% vs 15%; chi square = 4.5, P < 0.05; odds ratio 2.4; 95% CI 1.03-5.36). No association of the Pro12Ala variant with any of the component of the metabolic syndrome measured in the study was observed in either obese, juvenile obese or normal weight participants. CONCLUSIONS: Results of this study indicate that the Pro12Ala mutation does not play a major role as a determinant of severe obesity and/or features of the metabolic syndrome in the general population. However, this mutation may be of greater importance as a contributor to early onset obesity. PMID- 11033991 TI - Estimation of body fat in Caucasian and African-American girls: total-body electrical conductivity methodology versus a four-component model. AB - BACKGROUND: Obesity has been increasing dramatically in recent years among children, particularly African-American girls. Total-body electrical conductivity (TOBEC) is a simple way to measure body fat with minimal risk. OBJECTIVE: This study compared the agreement between the percentage of fat mass (%FM) predicted using two TOBEC equations with %FM measured by a four-component model in 73 Caucasian and 41 African-American girls. DESIGN AND MEASUREMENTS: %FM predicted using the TOBEC equations was compared with %FM from the four-component model based on measurements of body density, body water and bone mineral content. RESULTS: Analyses by linear regression analysis and by the Bland and Altman methods comparison procedure showed that the equation using the square root of the TOBEC zero-order Fourier coefficient and the subject's height yielded more accurate and more reproducible %FM, regardless of race, than the TOBEC linear equation, which was based on the zero-, first- and second-order Fourier coefficients. The Bland and Altman comparison further revealed that the accuracy and limits of agreement of the TOBEC linear equation were related to body fatness among the Caucasian girls. The relationship, however, disappeared when prepubescent girls and a girl with low %FM were excluded from the analysis. CONCLUSIONS: The TOBEC square root equation with adjustment for body geometry and length is recommended for use in adolescent girls, as it yielded better agreement with the criterion method. However, further validation of the TOBEC instrumentation for estimating body fat in prepubescent children and children with low body fat is warranted. PMID- 11033992 TI - Moderate-intensity regular exercise decreases serum tumor necrosis factor-alpha and HbA1c levels in healthy women. AB - OBJECTIVE: To evaluate the effects of moderate-intensity regular exercise on serum levels of tumor necrosis factor-alpha (TNF-alpha) and glucose and lipid metabolism parameters. DESIGN: Longitudinal intervention study of a 5 month exercise training program (30-45 min/day, 4-5 days/week). SUBJECTS: Forty-one healthy Japanese women aged 41-69 y at baseline; 27 participants in the exercise program. MEASUREMENTS: Body mass index (BMI), waist-to-hip ratio (WHR), percentage body fat, and fasting levels for serum TNF-alpha, serum soluble TNF receptor p55 (TNF-RI) and TNF receptor p75 (TNF-RII), serum lipids, HbA1c, and serum insulin before and after exercise. RESULTS: In overweight to obese subjects, serum levels of TNF-alpha, TNF-RI and TNF-RII were significantly higher than those in lean subjects. There were significant correlations between log serum TNF-alpha and BMI, percentage body fat, WHR, HbA1c and log insulin. TNF-RI was significantly correlated with BMI, percentage body fat, WHR and HbA1c. TNF RII was also associated with BMI, percentage body fat and WHR. However, the correlation between TNF-RII and HbA1c did not reach statistical significance. Neither TNF-RI nor TNF-RII was correlated with log insulin. In contrast, TNF alpha, TNF-RI and TNF-RII were negatively correlated with HDL cholesterol. Regular exercise decreased BMI, percentage body fat, HbA1c, serum TNF-alpha, TNF RI and TNF-RII and increased HDL cholesterol levels. In addition, exercise induced change in serum TNF-alpha was independently correlated with changes in HbA1c and serum insulin, after being adjusted for the change in fat-free mass. CONCLUSION: Changes in serum TNF-alpha that occur with exercise may play an important role in improving glucose metabolism parameters. PMID- 11033993 TI - High concentration of leptin stimulates myeloid differentiation from human bone marrow CD34+ progenitors: potential involvement in leukocytosis of obese subjects. AB - OBJECTIVE AND DESIGN: As well as its involvement in control of adipose mass and body energy balance, several reports suggest a link between leptin and hemopoiesis. To test its putative role in human hemopoiesis, we developed a homologous system, ie recombinant human leptin treatment of purified CD34+ progenitors from adult human bone marrow. RESULTS: Leptin (50-100 ng/ml) significantly stimulated the appearance of granulocyte-macrophage colonies in the presence or absence of erythropoietin. The concentration of leptin required for this effect was rather high but within the range of plasma leptin levels observed in obesity. Two results further support the hypothesis that leptin may be involved in the leukocytosis associated with obesity: (i) leptin concentrations in bone marrow and plasma of subjects studied were highly correlated; (ii) leptin and leukocyte count were correlated only in obese subjects. Paracrine effects of locally released leptin from bone marrow adipocytes could also be involved in the regulation of hemopoiesis, a hypothesis supported by marrow immunocytochemistry revealing the close association of CD34+ cells with adipocytes and by previous demonstration that leptin is secreted at a high level by these cells. CONCLUSION: These results indicate that leptin acts on human multilineage CD34+ cells and that high plasma leptin levels associated with obesity could participate in the differentiation of granulocytes from hemopoietic progenitors. PMID- 11033994 TI - Body weight characteristics of subjects on asthma medication. AB - OBJECTIVE: Weight gain is a frequently documented side effect after long-term anti-inflammatory treatment with systemic corticosteroid drugs in patients with asthma. In recent years new types of inhaled corticosteroids have been introduced, which act locally and are more rapidly bio-transformed. Even such corticosteroids may have a detectable, clinically relevant systemic side effect on weight. The aim of this study is to investigate if there is any relationship between body weight and asthma medication. DESIGN: The relationship between asthma medication and body weight was analysed in two combined randomized samples of the adult Swedish population 16-60 y of age (n = 17,912). Multivariate logistic regression analyses were carried out to obtain estimates for (1) body mass index (BMI) indicating 'obesity' (BMI > 29.9 kg/m2) in men and women controlling for self-reported asthma medication, and (2) self-reported asthma medication controlling for BMI. In both cases we furthermore controlled for interview period, age, Swedish region, smoking habits, physical activities and level of education. RESULTS: We found no significantly higher odds for obesity in men (OR = 1.21 (0.55-2.64) or women (OR = 1.97 (0.89-4.38) on asthma medication compared to men and women with pharmacologically untreated asthma even after adjustment for smoking habits, physical activities, level of education and other related co-variables. However, we found significant positive associations between obesity and interview period, age and former smoking and inverse significant relationships with the degree of physical activity. We also found significantly higher adjusted odds for asthma, indicated by self-reported asthma medication, in women (OR = 2.74 (1.91-3.91)) but not in men (OR = 1.57 (0.96-2.56)) with BMI indicating 'obesity'. CONCLUSION: There is no strong evidence to suggest that modern pharmacological asthma treatment may contribute much to the development of obesity in either men or women on asthma medication. Adjustment for smoking habits, physical activities, level of education and other related co-variables have minor effects on these relationships. Obesity may still be an independent risk factor for asthma since we observed significantly higher odds for self reported asthma medication in women and an almost significant relationship in men even after control for BMI and other related co-variables. PMID- 11033995 TI - Effects of weight variability on cardiovascular risk factors; a study of nonsmoking Japanese male office workers. AB - OBJECTIVE: To determine the effects of weight variability on cardiovascular risk factors. DESIGN: Longitudinal study (followed from 1990 to 1998). SUBJECTS: Five hundred and eighty nonsmoking Japanese male office workers aged 25-49 y. MEASUREMENTS: Baseline levels and slopes of seven selected cardiovascular risk factors (systolic and diastolic blood pressure, total and high-density lipoprotein cholesterol, triglyceride, uric acid and hemoglobin A1c) and body mass index (BMI) variables (baseline BMI, BMI-slope, the root mean square error (BMI-RMSE), and the BMI-slope x BMI-RMSE interaction). RESULTS: From the multiple regression analyses, the slopes of the six cardiovascular risk factors, except hemoglobin A1c, were most strongly related to the baseline level of each cardiovascular risk factor and BMI-slope. Neither BMI-RMSE nor the interaction of BMI-RMSE with BMI-slope was related to these cardiovascular risk factor slopes. As for hemoglobin A1c, BMI-slope and the interaction of BMI-RMSE with BMI-slope were not significantly related to hemoglobin A1c slope but baseline BMI and BMI RMSE were. Furthermore, hemoglobin A1c slope values were significantly higher among those who cycled at >4.0 kg than among those who did not. CONCLUSION: The only cardiovascular risk factor associated with BMI variability was hemoglobin A1c. Weight variability had less or little impact on cardiovascular risk factors compared with BMI-slope and baseline BMI. PMID- 11033996 TI - Transjugular intrahepatic portosystemic shunts in patients with hepatocellular carcinoma. PMID- 11033997 TI - Nutrition and enterocutaneous fistulas. AB - Enterocutaneous fistulas (ECFs) are a complex topic in terms of classification. ECF-related morbidity and mortality can be high due to fluid loss and electrolyte imbalance, sepsis, and malnutrition. Most prognostic factors influencing the outcome of ECF are now well-known. ECF treatment is complex; and, based on various situations, it can be surgical or conservative/ medical. Depending on fistula site and nutritional status, clinicians have to decide whether total parenteral or enteral nutrition should be established. In cases where total parenteral nutrition alone for 7 days has failed to influence the high output fistulas, overall data support the use of adjuvant drug, somatostatin, or its synthetic analogue, octreotide. Somatostatin 250 microg/d and octreotide 300-600 microg/d have been tried along with total parenteral nutrition to decrease the healing time of ECFs and to reduce the number of complications. PMID- 11033998 TI - Establishing a clinical research practice. AB - A clinical research practice can provide numerous benefits to a clinical gastroenterology practice. These benefits include new learning opportunities, enhanced community image, increased referral base, and improved practice revenues for the practitioner, as well as new treatment opportunities for the patient. A deliberate, business-like approach with appropriate personnel and space is necessary for a successful clinical research practice. All personnel should have a thorough understanding of good clinical practices. Avenues for obtaining a research grant are available, even for novice investigators, if the site is patient and committed. The purpose of this review is to explore the positive as well as the negative aspects of starting a clinical research practice, the requirements for getting the site started, finding the first study grant, and tasks to be anticipated during the conduct of a study. PMID- 11033999 TI - Correlation of endoscopy and histology in the gastroesophageal mucosa in children: are routine biopsies justified? AB - Guidelines for obtaining biopsies during endoscopy in children are needed. The endoscopic evaluation may be considered deficient on many occasions if not accompanied by a histopathologic evaluation. A retrospective review of our endoscopic records and biopsies was undertaken to determine the correlation of the visualized endoscopic appearance and the histopathologic findings in the upper gastrointestinal (GI) tract in children. Over a 1-year period, 204 patients, all of whom had esophageal biopsies and 59 of whom had gastric biopsies as well, were evaluated by an upper GI endoscopy. Endoscopic findings included erythema, granularity, abnormal vascular pattern, friability, erosions, plaques, ulceration, and strictures. Histologic evaluation of biopsies was undertaken by one pathologist according to the presence of and type of cellular infiltrate and cellular morphologic abnormalities in the mucosa and submucosa where available. In this study, the correlation of endoscopic appearance with histology was rather limited in both the esophagus and the gastric mucosa. Low specificity and sensitivity of endoscopy in both locations (41% and 81% for the esophagus; and 43% and 86% for the gastric mucosa, respectively) illustrated the discrepancy. The overall accuracy of endoscopic evaluation in matching the histologic diagnosis was not more than two out of three (63.8%). No single endoscopic finding had a reliable correlation with histologic diagnosis but some had higher predictive value than others. Of the multiple indications for endoscopy in children, recurrent abdominal pain had the least diagnostic yield. Endoscopic appearance correlates poorly with histologic diagnosis in the gastroesophageal mucosa in children. Regardless of the appearance of the mucosa, routine biopsy during upper GI endoscopy in children should be encouraged. PMID- 11034000 TI - Density of Helicobacter pylori infection evaluated semiquantitatively in gastric cancer. AB - Helicobacter pylori infection may play a role in the development of gastric cancer; however, a quantitative evaluation of the density of H. pylori infection has not been reported previously in relation to the histologic stage and type of cancer. This study was designed to compare the density of H. pylori infection to the histologic stage and type of gastric cancer. Between March 1996 and March 1998, surgical resection of primary lesion was performed in 50 patients with gastric cancer (39 men and 11 women with a mean age of 67 years) at our institution. Using immunohistochemical stains, the density of H. pylori infection was evaluated semiquantitatively at cancer site as well as noncancerous mucosa adjacent to cancer. This density was compared with the histologic stage and the type of gastric cancer. The severity of the mucosal atrophy was evaluated using the updated Sydney System. The prevalences and density of H. pylori infection decreased in proportion to advances in the cancer stage and the mucosal atrophy. In early cancer of the intestinal- and diffuse-type, the prevalence of H. pylori in adjacent sites was almost 90% and was significantly higher (p < 0.01) than that seen in the advanced cancer lesions. In the intestinal-type early cancer, the prevalence and density of infection was higher (p < 0.05) in the adjacent mucosa than in the cancer site, whereas in the diffuse-type early cancer, H. pylori was found in all cases at the cancer site and the adjacent site. In advanced cancer, the prevalence of H. pylori was about 40% in the adjacent site and about 10% in the cancer site in both histologic types. These figures were significantly lower (p < 0.01) than in the early cancers. The prevalence and density of infection did not differ in the intestinal- and diffuse-type gastric cancers, but did decrease with more advanced cancer stages. The changes in local environment of the advanced cancer may not be conducive to the survival of H. pylori. Thus, the prevalence of H. pylori may be affected by the histologic stage rather than the histologic type of gastric cancer, and the organism may play a similar role, but through different pathways, in the pathogenesis of both types of cancer. PMID- 11034001 TI - Rifabutin-based triple therapy after failure of Helicobacter pylori eradication treatment: preliminary experience. AB - Despite continuous improvement of Helicobacter pylori (Hp) eradication therapy, new treatment regimens are necessary if established first-line treatments fail. In the present pilot study, a recently described rifabutin-based triple therapy was evaluated after preceding failure of triple therapy. Rifabutin (150 mg), amoxicillin (1 g), and lansoprazole (30 mg) were administered twice daily for 1 week to 25 patients infected with Hp who had previously failed to respond to eradication treatment and/or who had developed resistance to macrolides and nitroimidazoles. Four patients were lost to follow-up. Eradication rate of rifabutin-based triple therapy was 86% (18/21; per protocol) and 72% (18/25; intention-to-treat). Side effects were minimal. It is concluded that this new drug combination is an effective therapy for Hp strains resistant to clarithromycin or metronidazole; however, rifabutin-based treatment regimens for Hp eradication should be restricted to patients infected with resistant strains. PMID- 11034002 TI - Paraduodenal hernia: a treatable cause of upper gastrointestinal tract symptoms. AB - Paraduodenal hernia (PDH) is an unusual condition that is caused by congenital intestinal malrotation. Noncatastrophic presenting symptoms and their responses to surgery have not been well-characterized. Barium upper gastrointestinal (UGI) series and small bowel follow-up x-rays, performed from December 1995 to September 1996, were sequentially reviewed by one radiologist (J.M.) to identify patients with small bowel series compatible with a PDH. Case histories were reviewed for symptomatic presentation, associated evaluation, and treatment. Based on the 294 UGIs and small bowel follow-throughs performed during this 10 month period, 6 cases were suspected to have a PDH. A right PDH was confirmed in the three patients who underwent surgical exploration (prevalence 1%). Preoperative patient symptoms included nausea, bilious vomiting, and right upper quadrant pain. Repair of the hernia defect resulted in complete resolution of chronic symptoms. Preoperative upper endoscopy, performed in three patients, was not helpful in identifying the disorder. Preoperative computerized tomography obtained in two patients was diagnostic for a right PDH. One symptomatic patient with vomiting and gastric stasis did not have surgery because of a terminal illness. The remaining two patients had no symptoms attributable to PDH. Patients with PDH frequently have chronic UGI symptoms. An upper endoscopy cannot be used to exclude this entity. After surgery, UGI symptoms from PDH are likely to resolve. PMID- 11034003 TI - Factors affecting gallstone recurrence after successful extracorporeal shock wave lithotripsy. AB - Ninety-six patients treated successively for symptomatic cholelithiasis with extracorporeal shock wave lithotripsy (ESWL) and oral bile acid therapy consisting of ursodeoxycholic acid in daily dosages of 600 mg were prospectively followed for gallstone recurrence for a median of 13 months. Ultrasonography was performed to detect stone recurrence at 3, 6, and 12 months, and then yearly after the termination of therapy. Recurrent stones were found in 17 patients (18%). The cumulative probability of gallstone recurrence was 15.8% at 12 months, 26.1% at 24 months, and 30.7% at 36 months. The probability of stone recurrence over the entire period of observation was not dependent on stone number, whereas the median interval to detection of recurrence was significantly shorter in the patients with multiple stones (2 months) than in those with solitary stones (8 months) (p < 0.05). The rate of impaired gallbladder contractility was higher in patients with recurrence (8/15, 53.3%) when compared with those with no recurrence (15/72, 20.8%) (p < 0.01). Neither age, gender, or stone characteristics predicted stone recurrence. Only one patient with a recurrence reported biliary pain. Of the 15 patients with recurrent stones who opted for further nonsurgical treatment, complete stone disappearance was achieved in 10. Impaired gallbladder function may predict gallstone recurrence after ESWL. PMID- 11034004 TI - Randomized comparative study of indocyanine green and India ink for colonic tattooing: an animal survival study. AB - Indocyanine green has been reported previously as a useful agent for colonic tattooing, but its durability, ease of use, cost, and safety have not been compared with India ink. Eight pigs were randomized to colonic tattooing using sterile indocyanine green or India ink injected through tandem filters. The animals were recovered and killed after 2 weeks and were examined. All animals survived the study period with no complications. We identified 17 of 29 (59%) indocyanine green tattoos and 48 of 52 (92%) India ink tattoos at autopsy (p = 0.0003). Histologic reactions were mild and were distributed similarly in each group and the intensity of the tattoos diminished similarly in both groups. Although indocyanine green was slightly more expensive, it was easier to handle compared with India ink. Both indocyanine green and India ink tattoos cause only mild histologic reaction in the swine colon when examined 2 weeks after injection. India ink is superior to indocyanine green, when used for colonic tattooing. PMID- 11034005 TI - Antiphospholipid antibodies are related to portal vein thrombosis in patients with liver cirrhosis. AB - The pathogenesis of portal vein thrombosis (PVT) in cirrhotic liver patients is not known. PVT has been related to liver dysfunction, neoplasm, hemodynamic factors, and hypercoagulability states. PVT has been reported in patients with antiphospholipid syndrome without liver cirrhosis. Our aim was to find the role of antiphospholipid antibodies (APAs) and coagulation inhibitors in PVT in patients with liver cirrhosis. We present a case-controlled study, matched by age, liver function, and etiology, to discover the role of APAs and anticoagulant protein activity in PVT in cirrhotic patients. We studied 30 cirrhotic patients: 6 of 10 (60%) patients with PVT were APA-positive, whereas only 2 of 20 (10%) in the cirrhotic control group were APA-positive (p < 0.005). Low serum levels of protein C, protein S antithrombin III, and plasminogen were found in cirrhotic patients; and, no differences were found between patients with and without PVT. Significantly lower protein S and antithrombin III levels were found in patients with Child-Pugh class C. Therefore, APAs were related to PVT in cirrhotic patients; but, a lower concentration of coagulation inhibitors was associated with liver dysfunction alone. PMID- 11034006 TI - Granular cells as a marker of early amiodarone hepatotoxicity. AB - Hepatotoxicity due to chronic amiodarone (AD) use is well described. However, hepatitis occurring after acute administration of AD has only occasionally been reported and the pathologic findings in the liver in this condition have not been well characterized. We describe an idiosyncratic reaction, in a 40-year-old man after 6 weeks of oral AD therapy, consisting of acute hepatitis, which resolved after withdrawal of the drug. The liver biopsy showed clusters of cells with granular cytoplasm. These cells were characterized as macrophages, and phospholipid membranous inclusions were demonstrated ultrastructurally in the granular cells and in the hepatocytes. Pathologists and clinicians should be aware of this subtle histologic finding when looking for evidence to support AD hepatotoxicity. PMID- 11034007 TI - Thrombotic thrombocytopenic purpura in acute pancreatitis. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare syndrome of unknown cause with an estimated incidence of one case per million. The disease is characterized by a pentad of symptoms: thrombocytopenia, microangiopathic hemolytic anemia, neurologic changes, renal dysfunction, and fever. It causes thrombosis in the microvasculature of several organs, producing diverse manifestations. Acute pancreatitis (AP) is a well-described consequence of TTP. We report a patient who developed TTP after presenting with AP, suggesting pancreatitis to be the cause, rather than a consequence, of TTP. PMID- 11034008 TI - Preoperative diagnosis of ileal lipoma by endoscopic ultrasonography probe. AB - The occurrence of tumor in the small intestine is relatively rare. It has been demonstrated that lipoma of the ileum is a cause of intussusception. We report a 59-year-old man admitted to our hospital for lower abdominal pain. Diagnosis of intussusception was made by abdominal x-ray and ultrasonography. Enema contrast studies revealed ileocolic intussusception. Colonoscopy revealed a tumor with an submucosal tumor (SMT)-like head and coil-spring appearance in the ascending colon. Endoscopic ultrasonography (EUS) revealed a hyperechoic submucosal lesion with features compatible with lipoma. Subsequently, this was confirmed histopathologically after resection. To our knowledge, this is the first report of preoperative diagnosis of ileal lipoma by EUS. PMID- 11034009 TI - Chemotherapy for hepatocellular carcinoma with portal hypertension due to tumor thrombus. AB - A case of hepatocellular carcinoma (HCC) complicated by tumor thrombosis of the main trunk is presented. Four courses of hepatic arterial infusion therapy, via a subcutaneously implanted injection port, were performed using cisplatin (10 mg for 1 hour on days 1-5) and 5-fluorouracil (250 mg for 5 hours on days 1-5). After four courses of the chemotherapy, marked reduction in size of HCC and the tumor markers were noted. The esophageal varices and ascites were improved after the chemotherapy with a recanalization of the left branch of the portal vein. The patient was doing well with a survival period of 28 months after the chemotherapy. These encouraging results suggested that the present therapy, based on the biochemical modulation, was a useful option for advanced HCC with portal hypertension due to tumor thrombosis of the main portal vein. PMID- 11034010 TI - Cetirizine-induce cholestasis. AB - Cetirizine, a human metabolite of hydroxyzine, is a selective H1-receptor antagonist currently approved for the treatment of seasonal allergic rhinitis, perennial allergic rhinitis, and chronic urticaria. In U.S. clinical trials, transient reversible hepatic transaminase elevations were observed in <2% of patients during cetirizine therapy. We report a case of cetirizine-induced cholestasis in a 28-year-old man with no previous hepatobiliary disease after a 2 year period of taking cetirizine on a daily basis. The treatment of this patient included the use of ursodeoxycholic acid, as well as hydroxyzine, for symptomatic relief of pruritus. In light of the patient's clinical and biochemical improvement while using hydroxyzine, it appears that the hepatic metabolism of hydroxyzine to metabolites, including cetirizine, is not involved in the pathogenesis of this particular case of drug-induced hepatotoxicity. Cetirizine should be considered as a potential cause of drug-induced cholestasis. PMID- 11034011 TI - Angioedema of the small bowel due to an angiotensin-converting enzyme inhibitor. AB - We describe a case of a 72-year-old woman who presented with two episodes of abdominal pain, vomiting, and diarrhea. Abdominal computed tomographic scans done during each episode demonstrated edema of the small bowel. Review of the patient's history revealed that she had been started on a treatment of lisinopril for hypertension 1 month before the first episode and had her prescribed dose increased 24 hours before each presentation. Angiotensin-converting enzyme (ACE) inhibitor-associated angioedema was suspected and the medication was discontinued. The patient has remained symptom-free while not taking the ACE inhibitor for 1 year. Review of the literature reveals only nine similar cases. All cases, including ours, occurred in women. Angioedema of the small bowel associated with ACE inhibitors is rare and often is not recognized before surgical exploration. Angioedema of the gastrointestinal tract should be considered in symptomatic patients taking ACE inhibitors. PMID- 11034012 TI - A case of hemangioma accompanied by inflammatory pseudotumor of the spleen. AB - Both hemangioma and inflammatory pseudotumor (IPT) of the spleen are rare benign mass lesions. Moreover, a splenic hemangioma accompanied by IPT is extremely rare. A 61-year-old woman who suffered from liver cirrhosis had a splenic cavernous hemangioma surrounded by granuloma. The literature on IPT of the spleen has described several possibilities of its causes; however, it is still unknown. This case was accompanied by portal hypertension due to liver cirrhosis, which may cause microrupture of hemangioma resulting in an IPT. PMID- 11034013 TI - Long-term observation after transjugular intrahepatic portosystemic stent-shunt in two patients with hepatocellular carcinoma. AB - Two patients with hepatocellular carcinoma (HCC) were treated with transjugular intrahepatic portosystemic stent-shunt (TIPS) and followed for 22 and 58 months thereafter. HCC was well controlled by transcatheter arterial chemoembolization. Hepatic failure or metastasis, especially in the lung, was not observed in the long-term observation. TIPS seems to be useful even in patients with HCC, provided HCC is controlled. PMID- 11034014 TI - Amphibian respiration and olfaction and their relationships: from Robert Townson (1794) to the present. AB - The present review examines the developments in the elucidation of the mechanisms of amphibian respiration and olfaction. Research in these two areas has largely proceeded along independent lines, despite the fact that ventilation of the nasobuccopharyngeal cavity is a basic element in both functions. The English naturalist Robert Townson demonstrated, in the 1790s, that amphibians, contrary to general belief, ventilated the lungs by a pressure-pump mechanism. Frogs and other amphibians respire by alternatively dilating and contracting the buccopharyngeal cavity. During dilatation, with the mouth and glottis closed, air is sucked in through the open nostrils to fill the cavity. During contraction of the throat, with nostrils closed and glottis open, the air in the buccopharyngeal cavity is pressed into the lungs. During expiration, the glottis and nostrils open and air is expelled from the lungs 'by their own contraction from a state of distention'. Herholdt (1801), a Danish army surgeon, independently described the buccal pressure-pump mechanism in frogs, his experiments being confirmed by the commissioners of the Societe Philomatique in Paris. Haro (1842) reintroduced a suction mechanism for amphibian respiration, which Panizza (1845) refuted: excision of the tympanic membranes prevented lung inflation, the air in the buccopharyngeal cavity leaving through the tympanum holes. Closure of the holes with the fingers restored lung inflation. The importance of cutaneous respiration in frogs and other amphibians was discovered by Spallanzani (1803), who found that frogs might survive excision of the lungs and that the amounts of exhaled carbon dioxide were small compared with those eliminated through the skin. Edwards (1824) confirmed and extended Spallanzani's findings, and Regnault & Reiset (1849) attempted to establish the relative importance of skin and lungs as respiratory organs in frogs. The problem was solved by Krogh (1904a) who measured respiration through the skin and lungs separately and simultaneously. Krogh (1904a) confirmed that carbon dioxide was chiefly eliminated through the skin, correlated with its high diffusion rate in water and tissue, whereas the pattern of oxygen uptake varied seasonally, the pulmonary uptake being lower than the cutaneous during autumn and winter, but substantially higher during the breeding period. Dolk & Postma (1927) confirmed this respiratory pattern. More recently, Hutchison and coworkers have examined the relative role of pulmonary and cutaneous gas exchange in a large number of amphibians, equipped with head masks for the separate measurement of the lung respiration in normally ventilating animals (Vinegar & Hutchison, 1965; Guimond & Hutchison, 1968; Hutchison, Whitford & Kohl, 1968; Whitford & Hutchison, 1963, 1965, 1966). As early as 1758, Rosel von Rosenhof suggested that the lungs of frogs in water functioned as hydrostatic organs that permitted the animal to float at the surface or rest on the bottom of the pond. The suggestion was inspired by observations made in the second half of the seventeenth century by members of the Royal Academy of Sciences in Paris. The French anatomists demonstrated that a tortoise, presumably the European freshwater turtle Emys orbicularis, could regulate its buoyancy by changing the volume of the lungs, to descend passively or ascend in the water. The hydrostatic function of the lungs has been repeatedly rediscovered, by Emery (1869) in the frog, by Marcacci (1895) in frogs, toads and salamanders, by Whipple (1906b) in a newt, by Willem (1920, 1931) in frogs and Xenopus laevis, by Speer 1942) in several anurans and urodeles, and finally by de Jongh (1972) in Xenopus laevis. In the second half of the nineteenth century a number of important papers appeared which confirmed and extended Townson's (1794) and Panizza's (1845) analysis of the normal respiratory movements in frogs. (ABSTRACT TRUNCATED) PMID- 11034015 TI - Trophic-dynamic considerations in relating species diversity to ecosystem resilience. AB - Complexity in the networks of interactions among and between the living and abiotic components forming ecosystems confounds the ability of ecologists to predict the economic consequences of perturbations such as species deletions in nature. Such uncertainty hampers prudent decision making about where and when to invest most intensively in species conservation programmes. Demystifying ecosystem responses to biodiversity alterations may be best achieved through the study of the interactions allowing biotic communities to compensate internally for population changes in terms of contributing to ecosystem function, or their intrinsic functional redundancy. Because individual organisms are the biologically discrete working components of ecosystems and because environmental changes are perceived at the scale of the individual, a mechanistic understanding of functional redundancy will hinge upon understanding how individuals' behaviours influence population dynamics in the complex community setting. Here, I use analytical and graphical modelling to construct a conceptual framework for predicting the conditions under which varying degrees of interspecific functional redundancy can be found in dynamic ecosystems. The framework is founded on principles related to food web successional theory, which provides some evolutionary insights for mechanistically linking functional roles of discrete, interacting organisms with the dynamics of ecosystems because energy is the currency both for ecological fitness and for food web commerce. Net productivity is considered the most contextually relevant ecosystem process variable because of its socioeconomic significance and because it ultimately subsumes all biological processes and interactions. Redundancy relative to productivity is suggested to manifest most directly as compensatory niche shifts among adaptive foragers in exploitation ecosystems, facilitating coexistence and enhancing ecosystem recovery after disturbances which alter species' relative abundances, such as extinctions. The framework further explicates how resource scarcity and environmental stochasticity may constitute 'ecosystem legacies' influencing the emergence of redundancy by shaping the background conditions for foraging behaviour evolution and, consequently, the prevalence of compensatory interactions. Because it generates experimentally testable predictions for a priori hypothesis testing about when and where varying degrees of functional redundancy are likely to be found in food webs, the framework may be useful for advancing toward the reliable knowledge of biodiversity and ecosystem function relations necessary for prudent prioritization of conservation programmes. The theory presented here introduces explanation of how increasing diversity can have a negative influence on ecosystem sustainability by altering the environment for biotic interactions and thereby changing functional compensability among biota- under particular conditions. PMID- 11034016 TI - The evolution of fitness in life-history theory. AB - Theory concerning the evolution of life history (the schedule of reproduction and survival) focuses on describing the life history which maximises fitness. Although there is an intuitive link between life history and fitness, there are in fact several measures of the 'black box' concept of fitness. There has been a debate in the bio-mathematical literature on the predictive difference between the two most commonly used measures; intrinsic rate of increase r and net reproductive ratio R0. Although both measures aim to describe fitness, models using one of the measures may predict the opposite of similar models using the other measure, which is clearly undesirable. Here, I review the evolution of these fitness measures over the last four decades, the predictive differences between these measures and the resulting shift of the fitness concept. I focus in particular on some recent developments, which have solved the dilemma of predictive differences between these fitness measures by explicitly acknowledging the game-theoretical nature of life-history evolution. PMID- 11034017 TI - Origin and evolution of primate social organisation: a reconstruction. AB - The evolution and origin of primate social organisation has attracted the attention of many researchers, and a solitary pattern, believed to be present in most nocturnal prosimians, has been generally considered as the most primitive system. Nocturnal prosimians are in fact mostly seen alone during their nightly activities and therefore termed 'solitary foragers', but that does not mean that they are not social. Moreover, designating their social organisation as 'solitary', implies that their way of life is uniform in all species. It has, however, emerged over the last decades that all of them exhibit not only some kind of social network but also that those networks differ among species. There is a need to classify these social networks in the same manner as with group living (gregarious) animals if we wish to link up the different forms of primate social organisation with ecological, morphological or phylogenetic variables. In this review, we establish a basic classification based on spatial relations and sociality in order to describe and cope properly with the social organisation patterns of the different species of nocturnal prosimians and other mammals that do not forage in cohesive groups. In attempting to trace the ancestral pattern of primate social organisation, the Malagasy mouse and dwarf lemurs and the Afro Asian bushbabies and lorises are of special interest because they are thought to approach the ancestral conditions most closely. These species have generally been believed to exhibit a dispersed harem system as their pattern of social organisation ('dispersed' means that individuals forage solitarily but exhibit a social network). Therefore, the ancestral pattern of primate social organisation was inferred to be a dispersed harem. In fact, new field data on cheirogaleids combined with a review of patterns of social organisation in strepsirhines (lemurs, bushbabies and lorises) revealed that they exhibit either dispersed multi-male systems or dispersed monogamy rather than a dispersed harem system. Therefore, the concept of a dispersed harem system as the ancestral condition of primate social organisation can no longer be supported. In combination with data on social organisation patterns in 'primitive' placentals and marsupials, and in monotremes, it is in fact most probable that promiscuity is the ancestral pattern for mammalian social organisation. Subsequently, a dispersed multi-male system derived from promiscuity should be regarded as the ancestral condition for primates. We further suggest that the gregarious patterns of social organisation in Aotus and Avahi, and the dispersed form in Tarsius evolved from the gregarious patterns of diurnal primates rather than from the dispersed nocturnal type. It is consequently proposed that, in addition to Aotus and Tarsius, Avahi is also secondarily nocturnal. PMID- 11034018 TI - Genes and plant cell walls: a difficult relationship. AB - Chemical information, carried by genes, is one of several types of information important for the functioning of cells and organisms. While genes govern the two dimensional flow of information, the cell walls are at the basis of a structural, three-dimensional framework of plant form and growth. Recent data show the walls to be a cellular 'organelle' undergoing dynamic changes in response to a plethora of stimuli. In this review, an integrated approach, rooted in the organismal perspective, is taken to consider the role of cell walls in the biology of plants. First, the complexity of molecular and biochemical events leading to the biosynthesis of wall components is described within the framework of its spatial cellular organisation, and the major regulatory check-points are characterised. Second, cell walls form a structural and functional continuum within the whole plant and thus could be defined in relation to the protoplasts that produce them and in relation to the plant itself. Model systems of suspension-cultured cells are used to reveal the existence of a bidirectional exchange of information between the protoplast and its walls. The 'plasticity' of plant cell reactions, seen in defence responses or in changes in wall composition, to e.g. stress, plant growth regulators or chemical agents as well as the role of cell walls and/or wall components in somatic embryogenesis are also discussed. Third, being a continuum within the plant body, the walls fulfil vital functions in plant growth and development. The examples characterised include the determination of cellular polarity and the plane of cell division, cytokinesis, and the role of plasmodesmata in cell-to-cell communication and the formation of functional symplastic domains. Fourth, the exocellular control of morphogenetic processes is described and the potential of cell walls as determinants or reservoirs of positional information is indicated. Particular emphasis is put on the (bio)chemical signals coming through or derived from cell walls as well as the mechanical properties of the walls. Based on those data, the 'plant body' concept is formulated. The plant is thus treated as a unit filled with intertwining networks: (1) symplastic, (2) the endomembrane system and (3) cytoskeletal, with cell walls providing an architectural scaffolding and communication ports formed within (4) the cytoskeleton-plasma membrane-cell wall continuum. PMID- 11034019 TI - Honors of the Association presented to Samuel Noordhoff, M.D. PMID- 11034020 TI - Distinguished Service Award presented to James A. Lehman, M.D. PMID- 11034021 TI - Craniofacial growth in unoperated craniofacial malformations. AB - OBJECTIVE: To review current information on craniofacial growth of unoperated patients with craniofacial malformations and stimulate the study of abnormal craniofacial growth through conventional and newer methods. This will lead to improved diagnosis and the understanding of the etiopathogenesis of craniofacial malformations. It is expected that this knowledge will also assist clinicians in planning treatment strategies to better manage these challenging conditions. PMID- 11034022 TI - The craniofacial dysostosis syndromes: current surgical thinking and future directions. AB - Craniofacial dysostosis is the term applied to familial forms of craniosynostosis in which the sutural involvement generally includes the cranial vault, cranial base, and midfacial skeletal structures. The syndromic forms of craniofacial dysostosis were initially described by Carpenter, Apert, Crouzon, Saethre and Chotzen, Pfeiffer, and others. In addition to the dysmorphic cranial features, affected individuals may have profound alterations in facial skeletal development. Surgical reconstruction requires thoughtfully sequenced and staged procedures with consideration for the individual's specific malformations, craniofacial growth patterns, and psychosocial needs. Management of the craniofacial dysostosis syndromes is surgical, but the indications and the timing, type, and effectiveness of each stage of reconstruction have not been well evaluated and remains as much an art as a science. This article reviews the specific characteristic clinical features of the craniofacial dysostosis syndromes and presents current philosophy and rationale for the staging of reconstruction. PMID- 11034023 TI - Treacher Collins syndrome: current evaluation, treatment, and future directions. AB - OBJECTIVE: Treacher Collins syndrome (TCS) is an inherited disorder in which there are general bilateral symmetric anomalies of the structures within the first and second branchial arches. In general, there is complete penetrance and variable expressivity of the trait. The craniofacial rehabilitation of a child with TCS is tailored to the extent of the deformities involved: the orbitozygomatic region, the maxillomandibular region, the nose, facial soft tissues, and external and middle ear structures. CONCLUSION: This article reviews the range of clinical features and specific dysmorphology observed in TCS. Functional and aesthetic objectives are discussed, and a comprehensive staged reconstructive approach is outlined, which may be used as a roadmap for treatment planning. PMID- 11034024 TI - Standardized facial photography of cleft patients: just fit the grid? AB - OBJECTIVE: The objective results of craniofacial surgery are usually evaluated by comparing pre- and postoperative photographs. To allow for optimal comparison, these photographs should be of constant and reproducible lay-out and quality. For this, various techniques to standardize photography have been suggested. DESIGN, SETTING, AND PATIENTS: A review of the literature on standardization of medical photography is presented. The advantages and disadvantages of standardizing techniques are evaluated. The difficulty to achieve standardization in cleft patients is illustrated using clinical photographs. Our personal protocol is presented as it tries to optimize comparability of photographs. RESULTS: Use of a grid or cephalostat will not lead to comparable photographs in cleft patients. Personal protocols also have shortcomings, but they offer reproducible and comparable results. A personal effort of the photographer is essential to adjust any allegedly standardizing protocol to the individual patient. CONCLUSION: Photographs are only as good as the photographer. True standardization of facial photographs is a utopia in cleft patients. PMID- 11034025 TI - Abnormal brain morphology in patients with isolated cleft lip, cleft palate, or both: a preliminary analysis. AB - OBJECTIVE: The aim of this study was to determine whether adult men with cleft lip and palate (CLP) have aberrant cerebral morphology. DESIGN: Brain morphology of 14 adult men with isolated CLP were analyzed and compared with 14 healthy controls matched for sex, age, and parental socioeconomic status. SETTING: The research took place at a large, tertiary care hospital, with participation on an outpatient basis. PARTICIPANTS: The 14 males with CLP were recruited from a large cleft lip and palate registry, while their 14 matched controls were selected from a registry of healthy volunteers collected via the Mental Health Clinical Research Center. RESULTS: The males with CLP have significantly smaller cerebellar size (p = .04), significantly larger frontal lobes (p = .02), and significantly smaller temporal and occipital lobes (p = .02; p = .009, respectively). No significant difference in gray/white matter ratios or laterality were found. CONCLUSIONS: Adult males with CLP have a significantly different pattern of brain morphology, compared with healthy controls, which is most likely due to aberrant cerebral development. This study highlights the complex interaction and interdependence of craniofacial and cerebral development. PMID- 11034026 TI - Dental health indices and caries associated microflora in children with unilateral cleft lip and palate. AB - OBJECTIVE: To investigate the dental health and caries related microflora of children with unilateral cleft lip and palate. STUDY GROUP: Sixty children with unilateral cleft lip and palate and matched controls. OUTCOME MEASURES: The decayed, missing, and filled teeth and surfaces in both the deciduous and permanent dentitions. The presence of developmental defects and plaque and gingivitis scores were also recorded. Plaque was collected from 25 of the children and their matched controls from three different sites, which were (1) the first approximal site distal to the cleft, (2) a contralateral anterior site, and (3) a remote site. It was cultured for Streptococcus mutans and lactobacilli. Plaque was collected from two sites in the matched controls. RESULTS: There was no significant difference in the caries, plaque, and gingivitis scores between the children with cleft palate and the controls. A greater number of enamel opacities were recorded in the control group, and there was a higher prevalence of enamel discoloration in the children with cleft lip and palate. There was no significant difference in the proportion of S. mutans or lactobacilli at the cleft site, compared with the unaffected site in the study group, although there was an anterior-posterior gradient in the proportion of S. mutans. There was no significant association between the stagnation area at the cleft site and the bacteria associated with dental caries. PMID- 11034027 TI - Dental arch relationships in Yorkshire children with unilateral cleft lip and palate. AB - OBJECTIVE: To assess the dental arch relationships of children with a complete unilateral cleft lip and palate (UCLP), born consecutively between 1983 and 1987, who had undergone primary cleft repair in the West Yorkshire region of the United Kingdom. The treatment outcome of this UCLP sample was then compared with the results of a previously published intercenter European study. DESIGN: A retrospective study with standardized record collection and blind assessment. SETTING: Regional dental hospital providing secondary and tertiary health care to the local population. PATIENTS: 35 UCLP patients whose primary repair had been performed within West Yorkshire. INTERVENTIONS: Dental casts obtained for each subject within 12 months of their 10th birthday. MAIN OUTCOME MEASURES: The West Yorkshire models were randomly mixed with an anonymous sample of 115 UCLP cases from other cleft centers in the U.K. The dental arch relationships of the 10-year old models were assessed by applying the Goslon Yardstick. RESULTS: One-third of the West Yorkshire cleft sample were rated as belonging to Goslon group 2 (good result), another third to group 3 (fair result), and the remaining third to groups 4 and 5 (poor/very poor result). Numerically, the results were of a slightly higher standard than that previously published for other U.K. and European cleft centers but were inferior to the treatment outcome of two European centers. No statistically significant differences were found between the outcome results of the West Yorkshire group and the other cleft centers. CONCLUSIONS: These results provide useful baseline data against which progress in achieving improved treatment outcome for West Yorkshire UCLP children can be determined by future research. PMID- 11034028 TI - Nasalance score variation in normal adult Japanese speakers of Mid-West Japanese dialect. AB - OBJECTIVE: The aims of this study were to examine nasalance score variation for normal adult Japanese speakers of Mid-West dialect and the gender difference in average mean nasalance score. DESIGN: Nasalance scores were obtained using a nasometer model 6200. The sample stimulus "Kitsutsuki passage," constructed of four sentences containing no Japanese nasal sounds, was used three times by each subject. PARTICIPANTS: One hundred normal adult speakers (50 women and 50 men) of Japanese served as subjects. The subjects ranged in age from 19 to 35 years of age (24.0 +/- 3.2). MAIN OUTCOME MEASURE: A mean nasalance score as well as an overall average nasalance value across speakers was calculated for each subject. The average mean nasalance scores between men and women were compared. RESULTS: The average mean nasalance score for the normal Japanese speakers was 9.1% (+/- 3.9). There was no statistically significant sex difference (p < .01). Average mean scores of 9.8% (+/- 3.5) and 8.3% (+/- 4.0) were obtained for the female and male speakers, respectively. CONCLUSIONS: The results provide important information concerning criteria to evaluate hypernasal speech due to velopharyngeal inadequacy of Japanese speakers with cleft palate using the nasometer. PMID- 11034029 TI - Pressure-flow characteristics of /m/ and /p/ production in speakers without cleft palate: developmental findings. AB - OBJECTIVE: The purpose of this study was to describe the pressure-flow characteristics of a large sample of speakers without cleft palate ranging in age from early childhood to young adulthood. METHOD: Speakers consisted of 223 children, teens, and adults without cleft palate categorized into five age groups: 6 through 8 years, 9 through 10, 11 through 12, 13 through 16, and 18 through 37 years. Speakers produced the syllables /mi/, /pi/, and /p/?/, the word "hamper," and the sentence "Peep into the hamper." The pressure-flow method was used to determine oral air pressure, nasal airflow, and estimates of velopharyngeal (VP) orifice size associated with /ml and /p/ production. Descriptive statistics were computed for each age group and speech sample. Analysis of variance (ANOVA) procedures were used to determine the effects of age, sex, and production level (word versus sentence) on the aerodynamic variables. RESULTS: ANOVA procedures indicated significant main effects (p < .01) of age on most of the aerodynamic variables during production of /m/ and /p/. No significant main effects or interactions involving sex were found for any variable. Regardless of age, approximately 95% to 99% of the speakers exhibited airtight VP closure during /p/ at syllable level, depending upon the selected nasal airflow criterion. ANOVA procedures also indicated significant main effects of production level (word versus sentence) on each of the aerodynamic variables during the /mp/ sequence. These effects appeared to be related to speaking rate. CONCLUSIONS: The study suggests that speakers without cleft palate exhibit essentially complete VP closure during production of oral pressure consonants in isolated syllables, and developmental aspects of speech aerodynamics be considered during pressure-flow testing. PMID- 11034030 TI - Effect of placement of a speech appliance on levator veli palatini muscle activity during speech. AB - OBJECTIVE: The purposes of this electromyographic study were to examine whether levator veli palatini muscle activity during speech can be changed with placement of a speech appliance and to clarify whether or not the change is related to the type of speech appliance used. DESIGN: Electromyography (EMG) was performed during production of speech samples in two conditions: with placement and with removal of a speech appliance. Speech samples were the vowel /omega/ and consonant-vowel (omega) syllables, including nasal, plosive, fricative, and affricate consonants. SETTING: Division for Oral-Facial Disorders, Faculty of Dentistry, Osaka University Dental Hospital, Japan. PARTICIPANTS: Subjects were six patients with repaired cleft palate and velopharyngeal inadequacy (VPI); three routinely wore a palatal lift prosthesis to correct borderline VPI, and the other three wore a speech appliance hybrid that consisted of a pharyngeal bulb and a palatal lift. INTERVENTIONS: Each patient was asked to produce each speech sample in both conditions of placement and removal of their respective appliances. MAIN OUTCOME MEASURES: Smoothed EMG signals of the levator veli palatini muscle were recorded with and without the appliance during five repetitions of each speech sample. The average value of peak smoothed levator EMG was compared between placement and removal conditions for all subjects and between the two subject groups in each experimental condition. RESULTS: With a speech appliance in place, the range of levator activity was distributed in a portion lower than that in the removed condition for subjects in both appliance groups. In addition, the intervals were smaller in the placement condition than in the removed condition. CONCLUSION: A speech appliance may prevent hypernasality, nasal emission of air, or both associated with VPI because of mechanical obturation of the velopharynx and an alteration in velopharyngeal function. PMID- 11034031 TI - Velopharyngeal closure pattern exerted by contraction of the pharyngeal constrictor muscle in dogs. AB - OBJECTIVE: This study was performed to investigate aspects of velopharyngeal closure demonstrated by movements of the parapharyngeal wall under various physiological conditions and especially the stimulation of several intrapneumonal receptors. MATERIALS AND METHODS: Fifteen mongrel dogs were anesthetized and a nasopharyngeal fiberscope was inserted from the oronasal fistula at the center of the hard palate, with the tip fixed around the choana. Mechanical and chemical stimulation was applied, and ongoing velopharyngeal movements were observed. In each case examined, the muscle nerve located under the bilateral levator veli palatini (LVP) was severed. RESULTS: Two different closure patterns were recognized between rebreathing and deglutition. Chemical stimulation of the pulmonary irritant receptor and type-J receptor induced a sphincteric tight closure of the velopharynx but only a weak inward movement of the lateral pharyngeal wall at lung inflation. CONCLUSION: The movements of the parapharyngeal wall, mainly represented by the pharyngeal constrictor muscle's contraction, are facilitated as a sphincteric velopharyngeal closure under reflexive movements such as deglutition or gagging. PMID- 11034032 TI - Analysis of scar tissue distribution on rat palates: a laser Doppler flowmetric study. AB - OBJECTIVE: The aim of this study was to evaluate the effectiveness of laser Doppler flowmetry (LDF) to discriminate postoperative scar tissue distribution on the palate. METHODS: Nineteen male Wistar rats at the 20th postnatal day were divided into experimental and control groups. In the experimental group, lateral palatal mucoperiosteum was excised to form scar tissue on the palate. At the 11th postnatal week, changes in the palatal blood flow were recorded with LDF in both groups by occluding exposed common carotid arteries. Perfusion values of nonoccluded (stable) and occluded states and the ratio of occluded to stable states were compared for scar tissue and normal tissue areas, and also for the normal tissue areas. After the LDF measurements, India ink-injected specimens and tissue sections were prepared for histological observations. RESULTS AND CONCLUSIONS: Scar tissue areas showed lower perfusion values both in the stable and occluded states, reflecting a lower vascular density in the scar tissue. The ratio of the occluded to stable states was higher in the scar tissue than in the normal tissue areas. In normal tissue areas, perfusion values of both the stable and occluded states appeared to vary, but the ratio did not vary among the areas. In the LDF study, the ratio of the occluded to stable states was considered to be the better parameter for discriminating scar tissue from normal tissue. PMID- 11034033 TI - Euclidean distance matrix analysis of surgical changes in prepubertal craniofacial microsomia patients treated with an inverted L osteotomy. AB - OBJECTIVES: Correction of craniofacial microsomia (CFM) presents several challenges concerning the modality of surgical intervention. The aim of this study was to assess early and late surgical outcome, by undertaking Euclidean distance matrix analysis (EDMA) of CFM patients exhibiting an unilateral mandibular deformity that was surgically corrected by an inverted L osteotomy and autogenous bone graft. DESIGN: Longitudinal study. Preoperative, approximately =1 year postoperative and approximately 3-year postoperative assessments of 14 consecutive children (mean age 9 years) with CFM. Posteroanterior cephalographs were scanned and five homologous mandibular landmarks were digitized in triplicate (< 1% digitization error). Average mandibular geometries, scaled to an equivalent size, were generated using a generalized rotational fit program (Procrustes superimposition) and subjected to EDMA. RESULTS: The mean pre- and both postoperative mandibular configurations differed statistically (p < .01). Early postoperative improvements in mandibular form were noted; increases in length arising in the treated mandibular body (approximately =19%) and ramus (approximately =13%). Comparing early and late postoperative configurations, a decrease of approximately =22% in the late postoperative mandibular body length was evident, but the ramus maintained steady vertical growth (approximately =7%). Comparing the preoperative and late postoperative configurations, the decrease observed in the mandibular body on the treated side was reduced to approximately =8% while the ramus maintained good growth (approximately =20%) on that side. CONCLUSION: Mandibular morphology is improved significantly in CFM patients surgically treated by an inverted L osteotomy, but relapse in the mandibular body is evident after approximately =3 years. Nevertheless, ramus growth proceeds well after the surgical reconstruction. PMID- 11034034 TI - Lipoma of the cleft soft palate: a case report of a rare congenital anomaly. AB - OBJECTIVE: Congenital tumors of the oral cavity are extremely uncommon. The teratoid tumors (epiganthi, dermoid, and hairy polyps) account for almost all of the reported cases and are not infrequently associated with cleft of the soft palate. Of the remaining tumors in the pediatric cleft palate population, infantile lipoma of the oral cavity is exceedingly rare. A case of a congenital lipoma associated with a cleft of the soft palate is presented. The review of the literature, the description of the lesion, the diagnosis, and the management of this finding are outlined. PMID- 11034035 TI - The effect of hamulus fracture on the outcome of palatoplasty: a preliminary report of a prospective, alternating study. AB - OBJECTIVE: To determine whether, in performing palatoplasty, fracture of the pterygoid hamulus is beneficial, detrimental, or neutral with respect to intraoperative and perioperative complications, hearing outcome, and speech outcome. DESIGN: Prospective, alternating. SETTING: Institutional, tertiary cleft palate center, Chang Gung Memorial Hospital, Taipei, Taiwan. PARTICIPANTS: A total of 173 patients enrolled in the study, of whom 161 had charts available for analysis. INTERVENTIONS: During the performance of palatoplasty, 85 patients received hamulus fracture and 76 patients did not. All palatoplasties were performed by the same surgeon. MAIN OUTCOME MEASURES: (1) Surgical outcomes, including patient demographic data, palatoplasty type and duration, blood loss, incidences of oronasal fistulae, temporary mucosal dehiscence, and postoperative bleeding; (2) otolaryngological outcomes, including hearing results as judged by auditory brainstem response testing, myringotomy tube data describing rates of tube extrusion, and culture results from sampled effusions; and (3) preliminary speech outcomes as described by judgments of overall velopharyngeal function from perceptual speech samples. RESULTS: No statistically significant differences in any of the measured surgical, otolaryngological, or preliminary speech outcomes were found between the groups who did and did not receive hamulus fracture. CONCLUSIONS: On the basis of these results, we are unable to advocate the performance of hamulus fracture as an operative maneuver during the performance of primary palatoplasty. The historical rationale and theoretical advantage of this maneuver have not been demonstrated here nor have any detrimental effects of the maneuver been measured. PMID- 11034036 TI - Major hematological diseases associated with cleft lip and palate. AB - OBJECTIVE: Cleft lip and palate is among the most common congenital anomalies. Its association with major blood disorders has rarely been reported. The purpose of this study was to report two patients who had major blood diseases associated with cleft lip and palate. PATIENTS AND RESULTS: From June 1995 to December 1997, there were 2700 patients with cleft lip, cleft palate, or both who received treatment at Chang Gung Memorial Hospital. Two of them were found to have major hematological disorders. In both cases, the disorder was detected by preoperative blood cell counts and white cell differentiation. Case 1 was a 21-year-old woman patient with repaired right cleft lip. She was admitted for alveolar bone grafting and closure of oronasal fistula. Abnormal presentation of blast cells was found, and subsequent bone marrow study confirmed acute lymphocytic leukemia. Case 2 was a 26-year-old man with left secondary cleft lip nasal deformity scheduled to receive staged reconstructive operations. An elevated platelet count was found and subsequently confirmed to represent essential thrombocytosis. In both cases, reconstructive operations for the cleft-related deformities were performed. CONCLUSIONS: Association of major hematological disorders and cleft lip, palate, or both is rare and is reported herein. PMID- 11034037 TI - An economic analysis of a randomized, controlled, multicenter study of clinical pharmacist interventions for high-risk veterans: the IMPROVE study. Impact of Managed Pharmaceutical Care Resource Utilization and Outcomes in Veterans Affairs Medical Centers. AB - STUDY OBJECTIVE: To determine if clinical pharmacists could affect economic resource use and humanistic outcomes in an ambulatory, high-risk population. DESIGN: Prospective, randomized, controlled study. SETTING: Nine Veterans Affairs medical centers. PATIENTS: Patients who were at high risk for medication-related problems. INTERVENTION: Patients were randomized to usual medical care with input from a clinical pharmacist (intervention group) or just usual medical care (control group). MEASUREMENTS AND MAIN RESULTS: Of 1,054 patients enrolled, 523 were randomized to the intervention group and 531 to the control group. The number of clinic visits increased in the intervention group (p=0.003), but there was no difference in clinic costs. Mean increases in total health care costs were $1,020 for the intervention group and $1,313 for the control group (p=0.06). CONCLUSION: Including the cost of pharmacist interventions, overall health care expenditures were similar for patients randomized to see a clinical pharmacist versus usual medical care. PMID- 11034038 TI - Randomized, placebo-controlled trial of oral phytonadione for excessive anticoagulation. AB - STUDY OBJECTIVE: To compare the efficacy of managing excessive anticoagulation in the absence of bleeding by either omitting warfarin therapy alone or administering oral phytonadione in addition to omitting warfarin therapy. DESIGN: Randomized, double-blind, placebo-controlled study. SETTING: Clinical pharmacy anticoagulation service in a group model health maintenance organization. SUBJECTS: Thirty nonbleeding patients with international normalized ratios (INRs) ranging from 6.0-10.0. INTERVENTIONS: Patients were randomized to receive either a single oral dose of phytonadione 2.5 mg or placebo. Both groups omitted warfarin doses until the INR became less than or equal to 4.0. MEASUREMENTS AND RESULTS: The mean calculated time to reach an INR of 4.0 was significantly greater in the placebo than the phytonadione group (2.6 vs 1.4 days, p=0.006). Overcorrection of anticoagulation was significantly more common in patients receiving phytonadione. Overt warfarin resistance was not observed in either group after reinitiating warfarin therapy. No major bleeding or thromboembolic complications occurred, and minor bleeding episodes were similar in both groups. CONCLUSION: The addition of oral phytonadione 2.5 mg reduced the time to achieve an INR of 4.0 by approximately 1 day compared with omitting warfarin therapy alone. Adverse events did not differ between the two groups. Both strategies were effective in managing asymptomatic patients with INRs of 6.0-10.0. Oral phytonadione may be most appropriate for patients at high risk for bleeding in whom the benefit of prompt INR reduction would outweigh the thromboembolic risk associated with INR overcorrection. PMID- 11034039 TI - Health-related quality of life for adults with cystic fibrosis: a regression approach to assessing the impact of recombinant human DNase. AB - STUDY OBJECTIVES: To examine the general health-related quality of life (HRQOL) of adults with cystic fibrosis (CF) using available generic instruments, and to determine important predictors of change in HRQOL, including therapy with recombinant human Dnase (rhDNase). DESIGN: Prospective, observational study. PATIENTS: Fifty-nine adults with CE. INTERVENTION: The Medical Outcomes Study Short-Form 36 (SF-36) and the EuroQol (EQ-5D) were mailed to patients; follow-up surveys were mailed 1 year later. MEASUREMENTS AND MAIN RESULTS: Multivariate regression analyses assessed the relationships between clinical and demographic variables and change in health status. The 39 respondents reported lower HRQOL scores compared with general population norms for generic instruments. Regression analyses indicated that age (p=0.010), percentage predicted forced expiratory volume in 1 second (p=0.005), hospital admission (p=0.023), and rhDNase therapy (p=0.026) were positively associated 1998 EQ-5D index scores, whereas body mass index was negatively associated (p=0.025) with 1998 EQ-5D index scores after controlling for baseline 1997 scores. The size of the regression coefficients indicated that only 1997 scores and hospital admission in the year were quantitatively importantly associated with 1998 scores. Baseline HRQOL assessments showed that adults with CF reported important decrements in health status and functioning. An interesting relationship between functional status and total household income was observed. CONCLUSION: Although a statistically significant association between rhDNase and 1-year change in HRQOL was found on multivariate regression analysis, the magnitude of this relationship may not be clinically important. PMID- 11034040 TI - Evaluation of the half-life of intravenous human cytomegalovirus immune globulin in patients receiving partially mismatched related donor bone marrow transplantation. AB - STUDY OBJECTIVE: To evaluate the pharmacokinetics and use of intravenous human cytomegalovirus immune globulin (CytoGam) in allogeneic bone marrow transplantation (BMT). DESIGN: Prospective, nonrandomized, nonblinded, single center study. SETTING: University teaching hospital. PATIENTS: Five consecutive patients with hematologic malignancies receiving partially mismatched related donor BMT with a uniform conditioning regimen including total body irradiation and chemotherapy. INTERVENTION: Serum immunoglobulin and cytomegalovirus (CMV) titers were measured before and 24 hours after the first CytoGam infusion on day 6 during the conditioning regimen. MEASUREMENTS AND MAIN RESULTS: These levels were measured every 5 days, and a second dose was administered when the CMV titer returned to 25-50% of the 24-hour level. The half-life of CytoGam was approximately 7 days. CONCLUSION: We believe this is the first report of CytoGam's half-life in allogeneic BMT. The information may prove vital in a future study in which the agent's potential beneficial effects can be maximized. PMID- 11034041 TI - Outcomes of weight-based heparin dosing based on literature guidelines and institution individualization. AB - STUDY OBJECTIVE: To determine whether unfractionated heparin is optimally dosed using published weight-based guidelines. DESIGN: Six-month, prospective study. SETTING: University hospital. PATIENTS: Ninety-six patients in the weight-based unfractionated heparin-dosing group 1 (WBHD1; 37 men; mean age 66.9 +/- 15.1 years; mean weight 80.1 +/- 20.6 kg) and 68 patients in the WBHD2 (25 men; mean age 68.2 +/- 15.6 years; mean weight 82.0 +/- 19.6 kg). INTERVENTIONS: The WBHD1 received a 100-U/kg intravenous bolus followed by an 18-U/kg/hour continuous intravenous infusion. After 3 months, the protocol was modified, and the WBHD2 received a 90-U/kg bolus followed by a 16-U/kg/hour continuous infusion for 3 months. MEASUREMENTS AND MAIN RESULTS: Activated partial thromboplastin times (aPTTs), frequency of bleeding episodes that required blood transfusions, and the number of recurrent thromboembolic events were collected from both groups after 3 months on the study. In the WBHD1, 24 hours after starting heparin, 38.5% of patients had therapeutic aPTTs, and at 48 hours, 54.3% were therapeutic. In the WBHD2, 42.6% and 51.4% of patients had therapeutic aPTTs at 24 and 48 hours, respectively. There was no statistical difference between the WBHD1 and WBHD2 in the percentage of patients with therapeutic aPTTs. CONCLUSIONS: Weight-based heparin dosing resulted in low percentages of patients with therapeutic aPTTs. The use of weight alone to dose heparin may not be adequate to optimize therapy. PMID- 11034042 TI - Hemodynamic and cardiovascular effects of nitric oxide modulation in the therapy of septic shock. AB - Nitric oxide synthase (NOS) of the inducible subtype (iNOS) plays a pivotal role in vasodilation associated with sepsis. Various biochemical pathways are involved, revealing targets for inhibiting the consequence of iNOS activation. Interactions of transcription factors, inducers, cofactors, and regulators of iNOS are important in understanding the development of iNOS inhibitors. Inhibition through L-arginine analogs, depletion of arginine, inhibition of cofactors, modulating gene transcription, and scavenging nitric oxide have been studied. Human studies were conducted only with nonselective L-arginine analogs. Reduction of mortality from sepsis was not reported. It is anticipated that iNOS specific compounds will be clinically useful. The focus of future human trials will be on these agents. Although ideal therapy for treating vasodilation from sepsis is not available, research into the pathophysiology of NOS in sepsis clarified the complexities surrounding this therapeutic dilemma. PMID- 11034043 TI - Clinical and economic effects of pharmacy services in geriatric ambulatory clinic. AB - Pharmacy services were introduced in an established multidisciplinary geriatric ambulatory clinic. The pharmacist collaborated with primary care providers to optimize patients' drug regimens. Over 8 months there were 250 patient visits to the clinic. Traditional medical care was provided at 144 (57.6%) of these visits and traditional medical care plus pharmacist evaluation was provided at 106 (42.4%). The pharmacist identified 220 potential and actual drug-related problems. Acceptance of pharmacist-recommended changes in drug therapy was 98.6%. A mean reduction of 3.4 agents/patient was achieved in the intervention group (p<0.0001). Clinical outcomes of changes in drug therapy were neutral or positive in 99.5% of cases. Pharmacy services resulted in net savings of $7,788 annually. PMID- 11034044 TI - Pharmacokinetics of gemcitabine and 2',2'-difluorodeoxyuridine in a patient with ascites. AB - Gemcitabine (dFdC) is a prodrug that undergoes metabolism by cytidine deaminase to form an inactive metabolite, 2',2'-difluorodeoxyuridine (dFdU). The pharmacokinetics of dFdC and dFdU have been studied; however, their disposition has never been evaluated in a patient with ascites. A patient with pancreatic cancer and malignant ascites was treated with dFdC 1,500 mg/m2 over 150 minutes weekly for 3 weeks, repeated every 4 weeks. Serial plasma and ascites samples were obtained on weeks 1 and 2 of cycle 2. High-pressure liquid chromatography was used to quantify dFdC and dFdU in plasma and ascites. The systemic dispositions of dFdC and dFdU were similar to those reported in patients without ascites. The concentration of dFdC in ascites approached 1 mg/ml. Ascitic fluid did not serve as a depot for dFdC, and the agent's concentration in ascites approached that at which its phosphorylation is saturated. PMID- 11034045 TI - Phytonadione therapy in a multiple-drug overdose involving warfarin. AB - We cared for a patient who ingested an unknown amount of acetaminophen with zopiclone and warfarin. The only liver function test that was abnormal was an increased international normalized ratio (INR), which remained elevated despite treatment with subcutaneous phytonadione and a prolonged infusion of N acetylcysteine. An interaction between acetaminophen and warfarin may have decreased the hepatic metabolism of warfarin. The patient received numerous antibiotics that may have contributed to the increased INR. The prolonged elevation of INR also may have been due to infrequent administration of phytonadione. PMID- 11034046 TI - Intravenous ribavirin and hyperammonemia in an immunocompromised patient infected with adenovirus. AB - Ribavirin is a synthetic guanosine analog with activity against DNA and RNA viruses. It was studied in human trials, and no marked adverse effect was reported beyond the potential for teratogenicity and reversible mild anemia. An 8 year-old girl received a multivisceral transplant and developed adenoviral pneumonia. She was treated with intravenous ribavirin and became hyperammonemic. Discontinuation of ribavirin led to a decrease in ammonia levels. This pattern was repeated when the drug was restarted and discontinued. We hypothesize that in a toxic environment the interaction of ribavirin with hepatocellular mitochondrial enzymes may lead to hyperammonemia. PMID- 11034047 TI - Palmar-plantar erythrodysesthesia syndrome associated with liposomal daunorubicin. AB - Daunorubicin and doxorubicin are anthracyclines that have efficacy against malignancies such as breast cancer, lung cancer, lymphoma, and leukemia. Their adverse effects are similar. The most serious is cardiotoxicity, which often limits the total cumulative dose that can be administered. Introduction of a liposomal formulation for both agents allows tumor selectivity by accumulating the drug in tumor tissue, thus increasing the tolerated dose. Liposomal doxorubicin is commonly associated with palmar-plantar erythrodysesthesia syndrome (PPES), although no reports of PPES were found in the literature related to liposomal daunorubicin (L-DNR). Two patients developed PPES while receiving high-dose L-DNR. The symptoms were self-limiting and resolved within a few weeks. PMID- 11034048 TI - Anagrelide-induced cardiomyopathy. AB - Anagrelide, a selective thrombocytopenic agent, is administered to treat a variety of hematologic disorders. Despite limited clinical experience with this drug, serious cardiovascular events, including congestive heart failure, have been reported. The proposed mechanism of cardiotoxicity is attributed to inhibition of phosphodiesterase, resulting in positive inotropic activity and vasodilation. A 48-year-old woman with polycythemia vera developed cardiotoxicity manifested by congestive heart failure and palpitations. It was suspected to be temporally related to titrating dosages of anagrelide. PMID- 11034049 TI - Clinical pharmacy: looking 20 years back... looking 20 years forward. PMID- 11034050 TI - Methods to assess the economic outcomes of clinical pharmacy services. AB - Studies that demonstrate the clinical and economic burden of drug misadventures have been published. The economic burden of drug misadventures likely exceeds $100 billion annually in the United States alone. Clinical pharmacy services (CPS) have been shown to reduce negative clinical outcomes and costs of drug therapy. Pharmacy practitioners and pharmacy managers need to demonstrate the impact of CPS at the local level and present the value to financial decision makers to gain support for implementation, continuation, and/or expansion of those services. The basic model of an economic evaluation includes measurement of both costs and consequences of a service and of an alternative for comparison. Strategies for economic assessments include the generalization from previously published literature, use of modeling techniques, and measurement of actual costs and consequences of an existing service. Guidelines for conducting an economic assessment are widely available. Pharmacy practitioners and managers should use these methods of assessing economic outcomes of CPS and, at the same time, develop relationships and skills to effectively communicate the value of those services so as to ensure long-term success. PMID- 11034051 TI - Methods to assess the humanistic outcomes of clinical pharmacy services. AB - This paper describes selected scientific principles involved in the design of patient self-administered questionnaires to assess functioning and overall well being. It gives several examples of successful uses of patient surveys and suggests possibilities for the use of the results by pharmacists in clinical care. The patients' point of view about their health status is important for the decision-making about treatment choices and as a monitoring parameter for effectiveness or toxicities. Pharmacists should approach the creation of new questionnaires or the results from untested questionnaires with caution and respect. The use of results from proven and reliable individual patient surveys will benefit from the evolution of computer adaptive testing. However, the immediate use of individual health status surveys is not discounted, provided that they accompany clinical data in decision processes. PMID- 11034052 TI - Technology for patient care assessment, documentation, and education. AB - Computer technology plays a big role in health care systems and information management. As technology continues to evolve, its role will become even more prevalent and important. Among the trends seen in technology products are better integration and compatibility of systems and accessories, enhanced performance and speed, smaller size and portability, improved user-friendliness, and affordability. All of these trends favor increased use in the health care environment. Selection of a product requires consideration of the desired goals and needs. An assessment should be made of existing technologies, particularly if interfacing will be required. Furthermore, most products have a life of only 10 18 months due to the rapid development of newer and better technologies. PMID- 11034053 TI - Clinical pharmacy in inpatient care. AB - The aim of this paper is to describe how interventions by clinical pharmacists can provide quality pharmaceutical care to inpatients. Evidence-based pharmacotherapy is necessary for the practice of clinical pharmacy. The use of published data, patient-specific data, and data generated by conducted research are discussed with respect to unit dose computer systems, nutrition teams, and pharmacokinetic services, based on the experience of an European hospital pharmacy. Incorporating evidence into clinical pharmacy activities results in added value to these services. The implementation of evidence-based pharmacotherapy is not an easy task for three reasons: a lack of scientific evidence in many medical areas, a lack of authority of pharmacists in Europe to take an active part in decision-making for drug prescribing, and reluctance of some physicians to accept pharmacists within the health care team. Also, efforts are required to change education programs at the universities in Europe to better prepare clinical pharmacists in pharmacotherapy and pharmaceutical care. PMID- 11034054 TI - Clinical pharmacy in disease-specific clinics. AB - The literature that describes and evaluates clinical pharmacy services in disease specific clinics is briefly reviewed. Various evaluation techniques are discussed, highlighting examples of sites using these techniques in published studies. The evaluation of clinical pharmacy services has evolved from examining a single parameter such as disease outcome to examining the broader perspective of the value of clinical pharmacists in the health care system, taking into account structure and process as well as outcomes. Assessment tools should consider all of these aspects to satisfactorily evaluate the value of clinical pharmacists in clinics. PMID- 11034055 TI - Clinical pharmacy in primary care and community pharmacy. AB - The role of the community pharmacist involved in primary care has been undergoing change. In the late 1970s and early 1980s, developments in computerized medication surveillance within the Netherlands enabled pharmacists to react to prescriptions and detect inappropriate pharmacotherapy in community pharmacy sites. This activity became more clinically or patient-oriented in the late 1980s. In the early 1990s, pharmaceutical care was introduced in community pharmacy practice, and emphasis was given to providing patient-centered care and documenting cognitive services. The key features of pharmaceutical care provided in the primary care setting are described based on a review of the literature and on experiences in the Netherlands. Patient outcomes have yet to be shown to be improved by community pharmacy practice; methods for measuring performance are still lacking. Methods to evaluate the extent of integration of community pharmacy services into the clinical team are also lacking but are needed in order to define the future role of community pharmacists in the primary care setting. Integrated care needs to be developed in the Netherlands in order to present the next phase in the process of the "pharmaceutical evolution." PMID- 11034056 TI - Curricular integration to enhance educational outcomes. AB - Integration of a pharmacy curriculum can enhance learning by structuring experiences so that students apply information and skills from a variety of domains-pharmaceutical sciences, clinical sciences, applied therapeutics, and practice experience-to solve problems. Successful curricular integration is accomplished by applying principles of instructional systems design and abilities based learning. Faculty from different disciplines must work together to focus on desired student performance, to view the curriculum as a system, and to assess students' abilities rather than their information recall. These approaches, while enhancing opportunities for effective learning and faculty collaboration, are difficult to design and implement. This paper discusses applying instructional design strategies to curricular integration, viewing the curriculum as a system, applying abilities-based learning strategies for curricular sequencing, and assessing student performance. PMID- 11034057 TI - Outcomes and experiential education. AB - Outcomes-based assessment in education involves continuous use of assessment measures to provide feedback about the efficacy of the curricular structure, content, and teaching methods. This process is initiated by establishing educational outcome statements for the Doctor of Pharmacy curriculum, selecting assessment methods that most appropriately measure the educational outcomes, and establishing a learning environment that is congruent with both the outcomes and assessment methods. To ensure a successful continuous outcomes-based assessment process, a systematic assessment plan should be prepared that focuses the process by identifying only the most essential hypotheses, uses a practical yet appropriate methodology, ensures efficient data collection, includes data analyses that link the educational outcomes to the learning environment, and promotes timely development and implementation of an action plan. An overview of outcomes-based education and the use of outcomes-based practice experiences in pharmacy education is presented. PMID- 11034058 TI - Effective teaching and learning strategies. AB - Over the past decade, leaders in pharmacy education and the profession in the United States focused on responding to the role of the pharmacist in the rapidly changing health care environment. This response included moving to the Doctor of Pharmacy as the entry-level degree in the profession. Less understood and often overlooked are pharmacy's responses to equally dramatic changes in the national higher education environment. One trend in higher education is the use of active teaching and learning methodologies, many of which are based on cognitive science research on human learning. The primary purpose of this paper is to focus on teaching and learning by describing the philosophy and policy changes of the new American Council on Pharmaceutical Education accreditation standards advocating active teaching and learning strategies, by discussing the implications for pharmacy faculty teaching roles and responsibilities, and by providing examples of active teaching and learning strategies and relevant literature embedded in the standards. PMID- 11034059 TI - Formulary management. AB - The hospital formulary is not only a list of drugs, but also a policy that involves several complex processes and activities. Formularies are developed based on cost-effectiveness evaluation, but maintaining cost-effectiveness requires educational strategies. The clinical pharmacist plays an active role in formulary processes. These processes are discussed in this paper. There is a need for studies on the cost-effectiveness of clinical pharmacy services in support of the hospital formulary. PMID- 11034060 TI - Target drug programs and medication use evaluation. AB - Target drug programs and medication use evaluations are activities that are undertaken to improve the correct use of drugs. These programs should focus on inappropriate drug use, drug use problems, optimizing use of drugs, and improving the level of patient care. To monitor the effects of the programs, several types of outcomes have been evaluated, such as economic and financial, clinical quality, quality of life, patient satisfaction, and collaborative practice. The methodology to classify and monitor drug use incorporates the classification system developed by the World Health Organization, which takes into account each drug's anatomic, therapeutic, and chemical classification. In order to avoid focusing only on drugs and drug costs in these programs, and to allow for monitoring the impact of the programs on clinical practice, linking drug data to patient data is stressed. Target drug programs improve the appropriate use of drugs, and by doing so, contribute to safe and rational use of drugs in society. PMID- 11034062 TI - How resource allocation decisions are made in the health care market. AB - This paper describes how economists view resource allocation decisions in health care markets. The basic economic decisions that must be made in any economic system and the resource allocation decisions in a perfectly competitive market are described. An idealized market can achieve an efficient allocation of resources and is contrasted with a more realistic description of the numerous ways in which health care markets depart from the perfectly competitive ideal. The implications of these departures for health care policy are discussed, along with key controversies concerning reliance upon markets for resource allocation in health care. In particular, the failure of competitive markets to achieve what many consider an equitable distribution of health care is emphasized. The paper concludes with some practical observations on how pharmacists can use the increasing emphasis on economic efficiency to the advantage of their profession. PMID- 11034061 TI - Assessing the value of pharmacists' health-systemwide services: clinical pathways and treatment guidelines. AB - Practice guidelines and clinical pathways are increasingly being used as tools to enhance the quality of health care services and to manage costs better. This article reviews the role of guidelines and clinical pathways in health care as defined within the broader concept of practice policies. The factors that increase the effectiveness of practice policies are examined. These include the origin of development, dissemination technique, and implementation strategy. Policies that are internally developed and implemented with concurrent reminder systems are the most effective. Clinical pathways fit these criteria and are therefore highly effective policy types. The roles that pharmacists within health systems can undertake in policy development are described. These include writing the policy document, providing expert review, providing education, and most important, facilitating the desired outcomes by implementing pharmacy services that promote compliance with the guidelines. Examples of pharmacy-based guideline and pathway implementation from the Henry Ford Health System are described for inpatient anticoagulation, outpatient preferred drug formulary policy, and outpatient lipid therapy management. PMID- 11034063 TI - How to develop and implement a program to continuously demonstrate the value of clinical pharmacy services. AB - This paper summarizes key elements that support the success of clinical pharmacy services to continuously improve the quality of drug therapy. Five drivers identify a successful model for continuously demonstrating the value of clinical pharmacy services: knowing the organizational culture; providing leadership; recruiting pharmacy sponsors; showing tenacity; and acting with management courage. Difficulties encountered when communicating the value of clinical pharmacy services within a health care organization usually arise because of failure to include one of these drivers. PMID- 11034064 TI - MYB oncogene amplification in hereditary BRCA1 breast cancer. AB - Comparative genomic hybridization analysis has demonstrated that breast tumors from BRCA1 and BRCA2 germ-line mutation carriers contain a large number of chromosomal copy number gains and losses. A high regional copy number gain at 6q22-q24 was observed in one BRCA1 tumor, and fluorescence in situ hybridization analysis indicated a strong amplification of the MYB oncogene (15 copies of MYB compared with 1 copy of chromosome 6 centromere). Fluorescence in situ hybridization analysis revealed amplification of MYB in 5 (29%) of 17 BRCA1 breast tumors, whereas none of 8 BRCA2 tumors and 13 breast cancer cell lines, and only 2 of 100 sporadic breast tumors exhibited altered MYB copy numbers. Gene amplification resulted in mRNA overexpression as determined by Northern blot and cDNA microarray analysis, and protein overexpression by immunohistochemical staining. We conclude that MYB amplification is infrequent in sporadic breast cancer but common in breast tumors from BRCA1 mutation carriers, suggesting a role of this cell cycle regulator and transcription factor in the progression of some BRCA1 tumors. However, we cannot rule out the significance of other genes in the 6q22-q24 amplicon. PMID- 11034065 TI - BRCA1 promoter region hypermethylation in ovarian carcinoma: a population-based study. AB - There is a clear association between germ-line BRCA1 mutations and inherited ovarian cancer; however, the association between BRCA1 mutations and sporadic ovarian cancer remains ambiguous. The frequency of BRCA1 promoter hypermethylation as an epigenetic means of BRCA1 inactivation was determined for a large, population-based cohort of ovarian cancer patients. BRCA1 promoter hypermethylation was determined by methylation-specific restriction digestion of tumor DNA, followed by Southern blot analysis and confirmed by methylation specific PCR. BRCA1 promoter hypermethylation was observed in 12 of 98 ovarian tumors. BRCA1 methylation status of the primary tumor was conserved in six recurrent tumors after interim chemotherapy. None of the 12 tumors with BRCA1 promoter hypermethylation demonstrated BRCA1 protein expression by immunohistochemistry. BRCA1 methylation was only seen in ovarian cancer patients without a family history suggestive of a breast/ ovarian cancer syndrome. Therefore, the 12 BRCA1 methylated tumors represented 15% (12 of 81) of the sporadic cancers analyzed in this study. Although the clinical significance of BRCA1 promoter hypermethylation is yet to be determined, promoter hypermethylation may be an alternative to mutation in causing the inactivation of the BRCA1 tumor suppressor gene in sporadic ovarian cancer. PMID- 11034066 TI - Blockade of nuclear factor-kappaB signaling inhibits angiogenesis and tumorigenicity of human ovarian cancer cells by suppressing expression of vascular endothelial growth factor and interleukin 8. AB - We determined whether blockade of nuclear factor (NF)-kappaB/relA activity in human ovarian cancer cells can suppress angiogenesis and growth in an orthotopic nude mouse model. The human ovarian cancer cells SKOV3ip.1 and HEY-A8 were transfected with a mutated IkappaBalpha (IkappaBalphaM), ie., resistant to phosphorylation and degradation, and hence blocks NF-kappaB activity. NF-kappaB signaling blockade significantly inhibited in vitro and in vivo expression of two major proangiogenic molecules, vascular endothelial growth factor and interleukin 8, in cultured cells and in cells implanted into the peritoneal cavity of nude mice. The decreased expression of vascular endothelial growth factor and interleukin 8 directly correlated with decreased tumorigenicity, decreased vascularization of lesions, decreased formation of malignant ascites, and prolonged survival of mice. These findings suggest that inhibition of NF kappaB/relA activity in ovarian cancer cells can suppress angiogenesis and progressive growth. PMID- 11034067 TI - Multiple genes at 17q23 undergo amplification and overexpression in breast cancer. AB - Studies by comparative genomic hybridization imply that amplification of the chromosomal region 17q22-q24 is common in breast cancer. Here, amplification and expression levels of six known genes located at 17q23 were examined in breast cancer cell lines. Four of them (RAD51C, S6K, PAT1, and TBX2) were found to be highly amplified and overexpressed. To investigate the involvement of these genes in vivo, fluorescence in situ hybridization analysis of a tissue microarray containing 372 primary breast cancers was used. S6K, PAT1, and TBX2 were coamplified in about 10% of tumors, whereas RADS1C amplification was seen in only 3% of tumors. Expression analysis in 12 primary tumors showed that RAD51C and S6K were consistently expressed in all cases in which they were amplified and also in some tumors without amplification. These data suggest that 17q23 amplification results in simultaneous up-regulation of several genes, whose increased biological activity may jointly contribute to the more aggressive clinical course observed in patients with 17q23-amplified tumors. PMID- 11034068 TI - Suppression of metastasis by thymidine phosphorylase inhibitor. AB - We developed a novel inhibitor of thymidine phosphorylase (TP), 5-chloro-6-[1-(2 iminopyrrolidinyl) methyl] uracil hydrochloride (TPI), that is about 1000-fold more active than 6-amino-5-chlorouracil, one of the most potent TP inhibitors. TPI inhibited the high chemotactic motility and basement membrane invasion of KB/TP cells, a TP-positive clone transfected with Rous sarcoma virus (RSV)/TP, to the levels seen in KB/CV cells, a control clone transfected with RSV. In nude mice, oral administration of TPI suppressed not only macroscopic liver metastases of highly metastatic KB/TP cells but also the level of human beta-globin as a molecular marker of micrometastases in the livers of the mice. These findings demonstrate that TP plays a key role in the invasiveness and metastasis of TP expressing solid tumors and suggest that TPI might be a novel antimetastatic agent for blood-borne metastasis. PMID- 11034069 TI - Paclitaxel induces release of cytochrome c from mitochondria isolated from human neuroblastoma cells'. AB - Paclitaxel is an antimicrotubule agent that induces mitotic block and apoptosis. We show for the first time that paclitaxel acts directly or mitochondria isolated from human cancer cells. In isolated yeast mito chondria, paclitaxel (15 microM) induced an 18% increase in the respiration rate, with no concomitant release of cytochrome c. In isolated neuroblas toma mitochondria, paclitaxel (10-100 microM) induced a 27-72% release o cytochrome c. Release was prevented by cyclosporin A, suggesting the involvement of the permeability transition pore. Doxorubicin did no induce cytochrome c release, whereas vinorelbine, another antimicrotu bule agent, did. Thus, antimicrotubule agents can directly affect mito chondria to induce apoptosis. PMID- 11034070 TI - Bcl-2 antisense oligodeoxynucleotide therapy of Epstein-Barr virus-associated lymphoproliferative disease in severe combined immunodeficient mice. AB - Bcl-2 is upregulated by Epstein-Barr virus (EBV) in immortalized lymphoblastoid (LCL) B cells and is expressed in the majority of EBV-associated posttransplant lymphoproliferative disorders (PTLDs). Given the antiapoptotic function and chemoprotective effects of Bcl-2, it represents a rational target for modulation using antisense oligodeoxynucleotides in Bcl-2-expressing, EBV-associated lymphoproliferative disorders. Using a fully phosphorothioated oligodeoxynucleotide targeted to the first six codons of Bcl-2, we examined the effects of Bcl-2 antisense both in vitro in LCLs and in vivo in the human/severe combined immunodeficient chimeric model of EBV-associated lymphoproliferative disorders. In vitro treatment of LCLs with Bcl-2 antisense in the presence of cationic lipid was associated with decreased expression of Bcl-2 protein, inhibition of proliferation, and stimulation of apoptotic cell death; these effects were sequence-dependent. Furthermore, treatment of LCL-bearing severe combined immunodeficient mice with Bcl-2 antisense but not control oligodeoxynucleotides completely prevented or significantly delayed the development of fatal EBV-positive lymphoproliferative disease in vivo. These studies demonstrate that Bcl-2 antisense oligodeoxynucleotides mediate sequence dependent antitumor effects in EBV-associated B-cell lymphoproliferations both in vitro and in vivo. These findings suggest that Bcl-2 antisense therapy may represent a novel antitumor treatment strategy for EBV-associated PTLDs and other Bel-2-expressing, EBV-positive malignancies. PMID- 11034071 TI - Tumor-specific transgene expression from the human telomerase reverse transcriptase promoter enables targeting of the therapeutic effects of the Bax gene to cancers. AB - Human telomerase reverse transcriptase (hTERT) is the catalytic subunit of telomerase, which is highly active in immortalized cells and >85% of human cancers but is quiescent in most normal somatic cells. To test the feasibility of using the hTERT promoter to induce tumor-specific transgene expression in cancer gene therapy, we constructed an adenoviral vector expressing a LacZ reporter gene driven by the hTERT core promoter and evaluated its activity in vitro and in vivo. The hTERT promoter could drive high-level expression of LacZ in tumor cells but not in normal cells and normal mouse tissues. Using a binary adenoviral system that can induce Bax gene expression, we showed that induction of the Bax gene expression via the hTERT promoter elicited tumor-specific apoptosis in vitro, suppressed tumor growth in nude mice, and prevented the toxicity of the Bax gene in vitro and in vivo. Thus, the hTERT promoter is apparently a strong and tumor-selective promoter with potential application in targeted cancer gene therapy. PMID- 11034072 TI - High frequency of chromosome 3p deletion in histologically normal nasopharyngeal epithelia from southern Chinese. AB - We have examined the presence of loss of heterozygosity (LOH) on chromosome 3p in histologically normal nasopharyngeal epithelia (NP), dysplastic lesions, and carcinoma of the nasopharynx from different ethnic and geographic regions. Microdissected normal NP from noncancerous individuals and nasopharyngeal carcinoma (NPC) samples from both the high-risk group (southern Chinese in Hong Kong) and two low-risk groups for NPC (central/northern Chinese in Anhui/Beijing and Caucasians in Toronto) were included. All NPC samples showed high incidence of 3p deletion (81-100%). High frequencies of LOH on 3p were also detected in normal NP (73.9%) and dysplastic lesions (75%) from the southern Chinese. Significant lower frequency of LOH on 3p was noted in normal NP from the low-risk groups (20%) than those from high-risk groups (P = 0.0003). The presence of such genetic alterations in the histologically normal NP and dysplastic lesions suggests that it is an early event in tumor development. The higher frequency of 3p LOH found in normal NP from southern Chinese compared with those from low-risk groups may be related to the distinct cancer incidence among these populations. PMID- 11034073 TI - 17q23 amplifications in breast cancer involve the PAT1, RAD51C, PS6K, and SIGma1B genes. AB - Amplification of the 17q23 region occurs frequently in breast tumors. To characterize the structure of 17q23 amplicons and to identify oncogene targets associated with this alteration, we performed a copy number analysis of 87 17q23 localized expressed sequence tags in seven breast cancer cell lines. Three major regions of amplification were detected in the MCF7 and BT474 cell lines. Amplification of at least one of four known genes (PAT1, PS6K, RAD51C, and SIGMA1B) was detected in the cell lines and in 28% of 94 breast tumors. In most cases, these four genes were overexpressed when amplified, but there was a particularly good association between amplification of the SIGMA1B gene and elevated expression in tumors, which suggested a possible role for this gene in tumor progression. Our data show that this region contains at least four independent targets of amplification, which suggests that there is considerable variability in the structure of the 17q23 amplicon. PMID- 11034074 TI - Progesterone regulates human telomerase reverse transcriptase gene expression via activation of mitogen-activated protein kinase signaling pathway. AB - Emerging evidence indicates that sex steroid hormones regulate telomerase in target tissues. We have reported that estrogen activates telomerase through transactivation of the telomerase catalytic subunit, human telomerase reverse transcriptase (hTERT). Progesterone usually antagonizes estrogen action in reproductive organs, but the effect on telomerase remains unclear. In this study, we examine the effects of progesterone on the gene expression of hTERT in breast and endometrial cancer cell lines expressing progesterone receptor. Progesterone significantly induced hTERT mRNA expression within 3 h after exposure. This transient effect peaked at 12 h and then decreased. In contrast, exposure to progesterone for > 48 h antagonized estrogen effects and inhibited the estrogen induced activation of hTERT expression; the cyclin-dependent kinase inhibitor p21/Waf1/Cip1 plays an integral role in this inhibition. Thus, progesterone exerts diverse effects on hTERT mRNA expression in a time-dependent manner. We also found that the mitogen-activated protein kinase signaling pathway mediates both the short-term and long-term effects of progesterone on hTERT gene expression. These findings support the notion that hTERT gene is a target of both estrogen and progesterone. PMID- 11034075 TI - An approach to analysis of large-scale correlations between genome changes and clinical endpoints in ovarian cancer. AB - This report describes analyses of associations of genome copy number abnormalities in ovarian cancers with clinical features using genome-wide graphical and analytical procedures. These studies show that tumor grade is a better indicator of the extent of genomic progression than stage, that loss of chromosome 4 occurs preferentially in high-grade tumors, and that gains of 3q26 qter, 8q24-qter, and 20q13-qter occur frequently in low-grade and low-stage tumors and thus may be early events in ovarian cancer development. In addition, loss of chromosome 16q24 and a total number of independent genome copy number aberrations >7 are associated with reduced survival duration. The association of loss of 16q24 (D16S3026) with decreased survival duration was confirmed by quantitative PCR. Regions that frequently are abnormal and associated with altered survival duration are strong candidates for higher resolution analysis and gene discovery and may be useful markers for prediction of clinical outcome. PMID- 11034077 TI - The phosphatidylinositol 3-kinase/AKT signal transduction pathway plays a critical role in the expression of p21WAF1/CIP1/SDI1 induced by cisplatin and paclitaxel. AB - The cyclin-dependent kinase inhibitor p21WAF1/CIP1/SD11 (p21) plays a crucial role in DNA repair, cell differentiation, and apoptosis through regulation of the cell cycle. A2780 human ovarian carcinoma cells, which are sensitive to cisplatin and paclitaxel, express wild-type p53 and exhibit a p53-mediated increase in p21 in response to the chemotherapeutic agents. Here, we demonstrate that phosphatidylinositol 3-kinase (PI3K) and its downstream targets serine/threonine kinases AKT1 and AKT2 (AKT), are required for the full induction of p21 in A2780 cells treated with cisplatin or paclitaxel. Inactivation of the PI3K/AKT signal transduction pathway either by its specific inhibitor LY294002 or by expression of dominant negative AKT inhibited p21 expression but had no inhibitory effect on the expression of the proapoptotic protein BAX by cisplatin and paclitaxel treatment. In addition, overexpression of wild-type or constitutively active AKT in A2780 cells sustained the regulation of p21 induction or increased the level of p21 expression, respectively. Experiments with additional ovarian carcinoma cell lines revealed that PI3K is involved in the expression of p21 induced by cisplatin or paclitaxel in OVCAR-10 cells, which have wild-type p53, but not in OVCAR-5 cells, which lack functional p53. These data indicate that the PI3K/AKT signal transduction pathway mediates p21 expression and suggest that this pathway contributes to cell cycle regulation promoted by p53 in response to drug-induced stress. However, inactivation of PI3K/AKT signaling did not result in significant alteration of the drug sensitivity of A2780 cells, suggesting that the cell death induced by cisplatin or paclitaxel proceeds independently of cell protective effects of PI3K and AKT. PMID- 11034076 TI - Regulation of gli activity by all-trans retinoic acid in mouse keratinocytes. AB - Sonic hedgehog (Shh) signaling is essential for many normal developmental processes. The Shh signal is interpreted by the Gli transcription factors. Elevated Gli-1 expression has been associated with several neoplasms, including basal cell carcinoma. All-trans retinoic acid (RA) has strong effects on epidermal growth and differentiation and has been used for the treatment of various epithelial disorders. In this report, we show that RA can inhibit Gli activity in immortalized murine keratinocytes in a RA receptor-specific manner. This inhibition may occur, at least in part, through sequestration of the transcriptional coactivator cyclic AMP-responsive element-binding protein-binding protein and suggests a novel effect of retinoid excess on Shh signaling. PMID- 11034078 TI - Tamoxifen-induced enhancement of calcium signaling in glioma and MCF-7 breast cancer cells. AB - The antiestrogen tamoxifen is commonly used to treat breast cancer, but it also has therapeutic activity in several other types of cancer. Many of these tumors, including malignant gliomas, are estrogen receptor negative. Nonetheless, high concentrations of tamoxifen can directly reduce cell proliferation in some of these tumors and induce apoptosis. In this study, the role of tamoxifen in calcium signaling and calcium-induced cell death was studied in both malignant glioma cell lines and MCF-7 breast cancer cells. Tamoxifen potently increased the spatial expansion of calcium waves by 30-150% while significantly enhancing and prolonging agonist-induced calcium elevations. Furthermore, tamoxifen pretreatment accelerated calcium ionophore-induced death by more than 20 min, suggesting that tamoxifen lowered cellular resistance to calcium loads. In contrast to its potentiating of calcium signaling in tumors, tamoxifen had no significant effect on calcium signaling in cultures of primary astrocytes from either human or rat brain. This study demonstrates the existence of calcium signaling in breast cancer and glioma cells and identifies tamoxifen as a potential modulator of tumor-associated calcium signaling. PMID- 11034079 TI - The collection of the motile population of cells from a living tumor. AB - In this study, we report that needles containing chemoattractants can be used to collect the subpopulation of motile and chemotactic tumor cells from a primary tumor in a live rat as a pure population suitable for further analysis. The most efficient cell collection requires the presence of chemotactic cytokines, such as epidermal growth factor and serum components, and occurs with 15-fold higher efficiency in metastatic tumors compared with nonmetastatic tumors. Although tumor cells of the nonmetastatic tumors show a motility response to serum, they were not collected with high efficiency into needles in vivo in response to serum, indicating that additional factors besides motility are required to explain differences in cell collection efficiencies between metastatic and nonmetastatic tumors. The results reported here indicate that needles filled with growth factors and matrigel, when inserted into the primary tumor, can faithfully mimic the environment that supports invasion and intravasation in vivo. Furthermore, the results indicate that the same cell behaviors that contribute to chemotaxis in vitro also contribute to invasion in vivo. PMID- 11034080 TI - Measurement of DNA copy number at microsatellite loci using quantitative PCR analysis. AB - This report describes the development and validation of quantitative microsatellite analysis (QuMA) for rapid measurement of relative DNA sequence copy number. In QuMA, the copy number of a test locus relative to a pooled reference is assessed using quantitative, real-time PCR amplification of loci carrying simple sequence repeats. Use of simple sequence repeats is advantageous because of the large numbers that are mapped precisely. In addition, all markers are informative because QuMA does not require that they be polymorphic. The utility of QuMA is demonstrated in assessment of the extent of deletions of chromosome 2 in leukemias arising in radiation-sensitive inbred SJL mice and in analysis of the association of increased copy number of the putative oncogene ZNF217 with reduced survival duration in ovarian cancer patients. PMID- 11034081 TI - Endostatin inhibits endothelial and tumor cellular invasion by blocking the activation and catalytic activity of matrix metalloproteinase. AB - Here we report the inhibition of cellular invasion by a recombinant mouse endostatin and the possible mechanism of the inhibition. Endostatin significantly reduced endothelial as well as tumor cellular invasion into the reconstituted basement membrane in vitro. Gelatin zymographic analysis revealed that the activation of promatrix metalloproteinase-2 (proMMP-2) that was secreted from endothelial cells was blocked upon endostatin treatment. Studies with recombinant MMPs confirmed that endostatin inhibited proMMP-2 activation, mediated by both membrane-type 1 MMP and 4-aminophenylmercuric acetate. Furthermore, enzymatic assays using a peptide substrate demonstrated that endostatin inhibited the catalytic activities of both MMP-2 and membrane-type 1 MMP. Finally, coimmunoprecipitation experiments revealed that endostatin formed a stable complex with proMMP-2. These novel findings would, at least in part, explain the mechanism of the potent antiangiogenic and antitumor activities of endostatin. PMID- 11034082 TI - Ovarian tumors in rats induced by chronic 2,3,7,8-tetrachlorodibenzo-p-dioxin treatment. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a multispecies reproductive toxicant, and it has been recently classified by IARC as a known human carcinogen. Here, we report that TCDD promotes the development of ovarian tumors in an initiation-promotion model in female Sprague Dawley rats. Rats were initiated with diethylnitrosamine (DEN) or vehicle at 70 days of age. Starting 2 or 18 weeks after initiation, rats were exposed biweekly to TCDD at a daily average dose of 125 ng/kg/day for 14, 30, or 60 weeks continuously or for 30 weeks plus withdrawal periods of 16 or 30 weeks. Fifteen of 76 (20%) rats initiated with DEN and promoted with TCDD for various lengths of time developed ovarian sex cord-stromal tumors of Sertoli cell type, whereas no ovarian tumors developed in 86 rats used as vehicle controls or that received DEN alone or TCDD alone. The highest tumor incidence occurred in 6 of 14 rats (43%) after 60 weeks of continuous TCDD after DEN initiation. One of six rats developed a tumor by 30 weeks of exposure. Because most effects of TCDD can be attributed to its activation of the aryl hydrocarbon receptor (AhR), the presence and localization of AhR was determined in the rat ovary and in the ovarian tumors by reverse transcription-PCR, immunohistochemistry, and in situ hybridization. AhR was localized to oocytes, granulosa and thecal cells of growing follicles, surface epithelial cells, and epithelial cells lining single tubules in ovaries from adult control Sprague Dawley rats. Neoplastic cells in the ovarian tumors were also positive for both AhR message and protein. These results indicate that the ability of TCDD to cause ovarian tumors is dependent on initiation, length of promotion, and age of the animal when exposed and evaluated. The tumor type induced by TCDD in this experimental system is the same histological subtype as that reported from an early study of youngsters exposed during an industrial accident in Seveso, Italy. PMID- 11034083 TI - Reconstitution of endogenous interferon a by recombinant interferon in hairy cell leukemia. AB - Recombinant human IFN alpha (rhIFN-alpha) plays an important role in the treatment of hairy cell leukemia (HCL). However, the mechanisms leading to its beneficial effect are not completely clarified, and there is no information on IFN-alpha gene expression in this disease. Therefore, we investigated the pattern of IFN-alpha gene expression and protein production in HCL and their potential regulation by rhIFN-alpha. Blood samples from 10 patients with HCL and 8 healthy donors (HD) were investigated. Expression of IFN-alpha mRNA was assessed by reverse transcription-PCR analysis in peripheral blood mononuclear cells (PBMCs) under basal conditions and on induction with rhIFN-alpha and polyionosinic polycytidylic acid [poly(I.C)]. IFN-alpha concentrations in plasma and culture supernatants were measured by immunoassays, and intracellular IFN-alpha was evaluated by fluorescence-activated cell sorting analysis. Results showed that, in contrast to blood samples from HDs, freshly isolated PBMCs from un treated HCL patients did not express IFN-alpha mRNA, whereas IFN-alpha transcripts were found in patients who were under rhIFN-alpha therapy Plasma of untreated patients contained no, or extremely low levels of IFN-alpha as compared with plasma of treated patients and HDs. Ex vivo treatment of PBMCs with rhIFN-alpha or poly(I.C) resulted in a remarkable up-regulation of IFN-alpha at the mRNA and protein level. In HCL, however the amounts of IFN-alpha protein remained less than in HD. Inhibition of IFN-alpha transcription was found after exposure of PBMCs to serum fron untreated patients. Finally, a reduced capacity to produce IFN-alpha was found within B- cell, T-cell, and monocyte compartments in HCL patients which could be enhanced by rhIFN-alpha. The results demonstrate the ability, of rhIFN-alpha to up-regulate the expression of IFN-alpha gene and protein production and suggest that priming the production of endogenous IFN alpha is a critical step in the mechanism of action of rhIFN-alpha in HCL. PMID- 11034084 TI - Polyadenylate polymerase enzymatic activity in mammary tumor cytosols: A new independent prognostic marker in primary breast cancer. AB - Polyadenylate polymerase (PAP) is one of the enzymes involved in the formation of the polyadenylate tail of the 3' end of mRNA. High levels of PAP activity were associated with rapidly proliferating cells. Here we evaluate the prognostic value of PAP activity in breast cancer patients. PAP specific activity values were measured by a highly sensitive assay in the tumor cytosols of 228 women with primary breast cancer. The median follow-up period was 58 months. PAP specific activity values ranged from 2.1-39.4 units/mg protein in the breast tumor cytosols, and the activity was correlated with the level of expression of the antigen. An optimal cutoff value of 5.5 units/mg extracted protein was first defined by statistical analysis. PAP status was then compared with other established prognostic factors in terms of relapse-free survival (RFS) and overall survival (OS). PAP activity levels had a tendency to increase with tumor node-metastasis (TNM) stage and were higher in node-positive patients. Evaluation of the prognostic value of PAP was performed using univariate and multivariate analyses. Univariate analysis showed that PAP-positive patients had a less favorable prognosis for both RFS (relative risk (RR) = 2.35; P < 0.001] and OS (RR = 3.15; P < 0.001). PAP significantly added to the prognostic power for RFS (RR = 2.51; P = 0.0012) and OS (RR = 4.21; P < 0.001) in multivariate analysis, whereas patient age, tumor size, and nodal and ER status remained independent factors for predicting survival. When only node-negative patients were examined, PAP was found to be an independent factor for predicting RFS (RR = 3.68; P = 0.0032) and OS (RR = 4.81; P < 0.001). PAP did not appear to have a prognostic significance for node-positive patients. PAP is a new prognostic factor for early recurrence and death in breast cancer patients. Our results suggest that PAP may be used as an independent unfavorable prognostic factor in node-negative breast cancer patients because there were no significant associations between PAP and the other prognostic indicators evaluated in this group of patients. PMID- 11034085 TI - Effects of dietary folate on intestinal tumorigenesis in the apcMin mouse. AB - Dietary folate appears to be inversely related to colorectal cancer risk. This study investigated the effects of dietary intervention with folate or the development of intestinal polyps in Min (Apc +/-) mice. Weanling Mil mice were fed diets containing 0, 2 (basal requirement), 8, or 20 mg folate/kg diet. At 3 and 6 months of dietary intervention, 50% of the mice from each group were sacrificed, and the small intestine and colon were analyzed for polyps and aberrant crypt foci (ACF). Serum folate concentrations accurately reflected dietary folate levels (P < 0.001). At 3 months no significant difference in the average number of total small intestinal polyps was observed among the four groups. However, increasing dietary folate levels significantly reduced the number of ileal, but not duodenal or jejunal, polyps in a dose-dependent manner (P-trend = 0.001); folate supplementation at 20 mg/kg diet was associated with a 68-78% reduction in the number of ileal polyps compared with the other three diets (P < 0.007). The number of ileal polyps was inversely correlated with serum folate concentrations (P = 0.03). At 3 months, increasing dietary folate levels significantly decreased the number of colonic ACF in a dose-dependent manner (P = 0.05); the control and two folate supplemented diets significantly reduced the number of colonic ACF by 75 100% compared with the folate-deficient diet (P < 0.04). The number of colonic ACF was inversely correlated with serum folate concentration (P = 0.05). No significant difference in the number of colonic adenoma was observed among the four groups at 3 months. At 6 months, no significant differences in the average number of total small intestinal, duodenal, and jejunal polyps, colonic adenomas, and colonic ACF wer observed among the four groups. However, the folate-deficient diet had 62-76% lower number of ileal polyps compared with the control and two folate-supplemented diets (P < 0.003). Serum folate concentrations, but not dietary folate levels, were directly correlated with the number of ilea polyps (P = 0.006). These data suggest that dietary folate supplementation suppresses the development of ileal polyps and colonic ACF in this model However, at later time points, folate supplementation appears to have an opposite effect on ileal polyps. These data generally support the role of folate in intestinal tumorigenesis suggested in epidemiological studies and chemical carcinogen animal models. Notwithstanding the limitations associated with this model, these data suggest that the optimal timing and dose of folate intervention need to be determined for safe and effective folate chemoprevention. PMID- 11034086 TI - Differential effects of estrone and estrone-3-O-sulfamate derivatives on mitotic. Arrest, apoptosis, and microtubule assembly in human breast cancer cells. AB - There is considerable interest in the potential use of estrogen derivatives for the treatment and prevention of breast cancer. We demonstrated previously that the sulfamoylated estrone derivative 2-methoxyestrone-3-O-sulfamate (2-MeOEMATE) induced G2-M cell cycle arrest and modest levels of apoptosis in breast cancer cells in vitro, whereas the parent estrone derivative, 2-methoxyestrone, did not. 2-MeOEMATE also induced breast tumor regression in vivo in intact rats. To further explore the significance of sulfamoylation on the anticancer activity of estrone derivatives and to elucidate their mechanism of action, we synthesized two additional agents, 2-ethylestrone and 2-ethylestrone-3-O-sulfamate (2EtEMATE). 2-MeOEMATE and 2-EtEMATE inhibited the growth of a panel of estrogen receptor-negative and -positive breast cancer cell lines in vitro, induced mitotic arrest and apoptosis, and suppressed the long-term clonogenic potential of MCF7 and CAL51 breast cancer cells. In each assay, the sulfamoylated estrone derivatives were >10-fold more potent than their parent compounds. The sulfamoylated estrone derivatives were also significantly more potent inhibitors of cell growth than the previously studied endogenous estradiol metabolite 2 methoxyestradiol. 2-MeOEMATE and 2-EtEMATE functioned as antimicrotubule agents and inhibited the ability of paclitaxel to promote tubulin assembly in vitro. Like other antimicrotubule agents, the sulfamoylated estrone derivatives induced BCL-2 and BCL-XL phosphorylation and increased p53 expression. 2-MeOEMATE and 2 EtEMATE are novel antimicrotubule agents that have potent anticancer activity in breast cancer cells in vitro and may be beneficial as anticancer agents in vivo. PMID- 11034087 TI - Inhibition of phosphatidylinositide 3-kinase enhances gemcitabine-induced apoptosis in human pancreatic cancer cells. AB - Human pancreatic adenocarcinoma cell lines PK1 and PK8 are resistant to the clinically relevant chemotherapy agent gemcitabine. Cell cycle analysis demonstrated an accumulation of cells in the early S phase during treatment with 20 microM gemcitabine, consistent with its mode of action as a DNA chain terminator. However, apoptosis was evident in only a small percentage of cells. Similar to pancreatic cancers in the clinic, PK1 and PK8 cells carry constitutively active Ki-Ras and overexpress multiple receptor tyrosine kinases. Both genetic abnormalities may potentially up-regulate the activity of the phosphatidylinositide 3-kinase (P13K)-protein kinase B (PKB)/Akt cell survival pathway. The current study examined the relevance of this pathway in the modulation of drug resistance in PK1 and PK8 cells. After exposure to 20 microM gemcitabine for 48 h and in the continuous presence of the drug, treatment with the P13K inhibitors wortmannin (50-200 nM) and LY294002 (15-120 microM) for 4 h substantially enhanced apoptosis in a concentration-dependent manner as compared with treatment with gemcitabine alone, as determined by the loss of mitochondrial membrane potential and the increase in propidium iodide uptake using flow cytometry. Furthermore, Western blotting showed that the reduction of phosphorylated PKB/Akt levels correlated with the enhancement of gemcitabine induced apoptosis, suggesting that the PI3K-PKB/Akt pathway plays a significant role in mediating drug resistance in human pancreatic cancer cells. PI3K inhibitors may have therapeutic potential when combined with gemcitabine in the treatment of pancreatic cancers. PMID- 11034088 TI - Adenoviral vectors targeted to CD40 enhance the efficacy of dendritic cell-based vaccination against human papillomavirus 16-induced tumor cells in a murine model. AB - Dendritic cells (DCs) represent a unique junction from which to initiate antigen specific immunity. One of the most challenging obstacles for DC-based immunotherapy has been the means by which to convey tumor antigen-encoding genes to DCs. In this study, we show that adenoviral (or adenovirus, Ad) vectors targeted to CD40 by means of bispecific antibodies can enhance gene transfer to murine DCs. Moreover, we illustrate that this vector initiates phenotypic changes characteristic of DC maturation. To explore the in vivo potential of this strategy, we coupled this targeting approach with an Ad vector carrying the gene for a tumor antigen. In particular, the human papillomavirus (HPV) E7 antigen represents an attractive target for antigen-specific immunity of cervical cancer. Relative to DCs infected by untargeted Ad, DCs infected by AdE7 targeted to the receptor CD40 enhanced protection against HPV-16-induced tumor cells in a murine model. We have further established that this protection was both antigen specific and CD8+ T-cell dependent. Illustrating that Ad-modified DCs may be used in repeated vaccination, we report that preimmunization of animals with Ad infected DCs prior to E7 vaccination only moderately reduced vaccine efficacy. Finally, we have observed that CD40-targeted AdE7 can initiate partial therapeutic immunity in mice bearing established tumors. These findings suggest that gene-based vaccination of DCs with tumor antigens can elicit productive antitumoral immunity and that enhancements in gene transfer efficacy and/or DC maturation may facilitate this process. PMID- 11034089 TI - Effect of O6-benzylguanine on alkylating agent-induced toxicity and mutagenicity. In Chinese hamster ovary cells expressing wild-type and mutant O6-alkylguanine DNA alkyltransferases. AB - The DNA repair protein O6-alkylguanine-DNA alkyltransferase (AGT) has been shown to protect cells from the toxic and mutagenic effect of alkylating agents by removing lesions from the O6 position of guanine. O6-Benzylguanine (BG) is a potent inactivator of AGT, resulting in an increase in the sensitivity of cells to the toxic effects of chemotherapeutic alkylating agents. Chinese hamster ovary (CHO) cells and CHO cells transfected with wild-type AGT (CHOWTAGT) and a mutant AGT [P138 M/V139I/P140K (CHOMIK)] known to be resistant to BG were treated with BG and various alkylating agents. BG treatment alone dramatically decreased AGT activity in CHOWTAGT cells but resulted in no depletion in AGT activity in CHOMIK cells. In the absence of AGT, these cells are highly sensitive to the toxic and mutagenic effects of temozolomide and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), and no further sensitization occurs in the presence of BG. In contrast, CHOWTAGT cells are resistant to temozolomide and BCNU, and treatment with BG resulted in a significantly higher cell killing and mutation frequency. CHOMIK cells were completely resistant to temozolomide or BCNU in the presence and absence of BG. Both cell killing and mutation frequency of 4 hydroperoxycyclophosphamide (4-HC) in CHO, CHOWTAGT, and CHOMIK cells were increased in the presence of BG. 4-HC generates two active metabolites, phosphoramide mustard (PM) and acrolein. BG had no effect on 4hydroperoxydidechlorocyclophosphamide (which generates acrolein and a nonalkylating form of PM) in CHO cells and CHOMIK cells, but enhancement of toxicity was observed with PM in both these cell lines. Therefore, we attribute the enhancement to the PM metabolite of 4-HC. Our results demonstrate that wild type AGT plays an important role in protecting against the toxic and mutagenic effect of O6 alkylating agents and that a mutant AGT resistant to inactivation by BG effectively prevents BG-enhanced toxicity and mutagenicity induced by these agents. Expression of the AGT protein contributes to resistance of 4-HC. BG also enhances the toxicity of 4-HC and PM by a mechanism that may not involve the AGT repair protein. PMID- 11034090 TI - Comparison of different busulfan analogues for depletion of hematopoietic stem cells and promotion of donor-type chimerism in murine bone marrow transplant recipients. AB - Busulfan (1,4-butanediol dimethanesulfonate, BU) is relatively unique among other standard chemotherapy compounds in its ability to deplete noncycling primitive stem cells in the host and consequently to allow for high levels of long-term, donor-type engraftment after bone marrow transplantation (BMT). Such a property explains why this drug can be used as an alternative to total body irradiation in preparative regimes for BMT. However, as with radiation, BU conditioning is still troubled by severe toxicities that limit its applications to suboptimal drug doses. These problems stress the need for other BMT-conditioning drugs that are better tolerated and more selectively targeted toward normal and malignant hematopoietic stem cells. We have therefore compared the effects of various novel dimethanesulfonate compounds (related to BU) in terms of their toxicity to different stem cell subsets in vivo and in vitro and their ability to provide for long-term donor bone marrow engraftment using the congenic glucose-6-phosphate isomerase type 1 marker. Introduction of a benzene or cyclohexane ring in some of these drugs affords rigidity to the molecule and restricts the spatial positioning of the alkylating groups. Among 25 different compounds thus far tested at single doses, PL63 [cis-1,2-(2-hydroxyethyl) cyclohexane dimethanesulfonate] proved to be the most effective in providing for hematopoietic engraftment. The transisomer of the same compound gave significantly less engraftment and was comparable with the effects of dimethylbusulfan and Hepsulfam. The engraftment data correlated well with the depletion of different bone marrow stem cell subsets in the host as measured using the cobblestone area forming cell assay. The extent of stem cell depletion could not be explained on the basis of the distance and orientation of the two alkylating groups. Pharmacokinetic data, however, indicate that there is a correlation between biological activity and plasma levels reached. The diverse cytotoxic effects shown by these novel analogues of BU have provided a basis for relating biological activity with pharmacokinetic properties rather than with structural properties such as distance and orientation of the two alkylating groups. The identification of highly active compounds such as PL63 offers an opportunity for further developing other closely related drugs for potential application in clinical BMT conditioning therapy. PMID- 11034091 TI - Activation of retinoic acid receptor alpha is sufficient for full induction of retinoid responses in SK-BR-3 and T47D human breast cancer cells. AB - Retinoid signaling via retinoic acid (RA) and retinoid X receptors (RARs and RXRs) regulates mammary epithelial cell growth and differentiation. Loss of RAR beta might represent an early event during breast carcinogenesis. Higher differentiated, estrogen-dependent, estrogen receptor (ER)-positive (ER+) mammary carcinoma cells have been found to contain relatively high levels of RAR-alpha and to be responsive to retinoids, whereas most undifferentiated, estrogen independent, ER-negative (ER-) cells are characterized by low RAR-alpha expression and by retinoid resistance. In contrast, RAR-gamma is detectable at equal levels in both ER+ and ER- cells. In the present investigation, we directly examined the relative contribution of the distinct retinoid receptors to the retinoid response of breast cancer cells by comparing the effects of low concentrations of specific retinoids, which selectively activate individual receptor subtypes, on growth, cell cycle distribution, apoptosis, and on the autoregulation of RAR-alpha and RAR-gamma in ER- SK-BR-3 and ER+ T47D breast cancer cells. In vitro growth activity was determined by using a colorimetric cell viability assay and analysis of cell cycle distribution, and apoptosis was performed by flow cytometry of propidium iodide-stained or fluorescent Annexin V labeled cells, respectively, whereas expression of RAR-alpha and RAR-gamma was determined by Northern blotting. Both cell lines are retinoid sensitive and express high amounts of RAR-alpha, RAR-gamma, and RXR-alpha. RAR-alpha-selective compounds (AM80 and AM580) inhibit cell growth, induce G1 arrest, stimulate apoptosis, and up-regulate RAR-alpha and RAR-gamma mRNA as efficiently as RAR/RXR pan-reactive (9-cis RA) and RAR-pan-reactive retinoids (all-trans RA, TTNPB). Remarkably, an RAR-alpha antagonist (Ro 41-5253) not only blocks the RAR-alpha selective agonists but also the pan-reactive compounds. In contrast, RAR-13 selective (CD417), RAR-gamma-selective (CD437/AHPN), and RXR-alpha-selective (Ro 25-7386) retinoids exert no effects on the examined parameters. Thus, our results support the idea that RAR-alpha is the crucial receptor mediating the biological effects during retinoid signaling in both ER- SK-BR-3 and ER+ T47D human breast cancer cells. PMID- 11034092 TI - Experimental cancer cachexia: the role of host-derived cytokines interleukin (IL) 6, IL-12, interferon-gamma, and tumor necrosis factor alpha evaluated in gene knockout, tumor-bearing mice on C57 Bl background and eicosanoid-dependent cachexia. AB - MCG 101 tumors were implanted sc. on wild-type C57 Bl and gene knockout mice to evaluate the role of host-produced cytokines [interleukin (IL)-6, IL-12, IFNgamma, tumor necrosis factor (TNF) receptor 1, and TNF receptor 2] to explain local tumor growth, anorexia, and carcass weight loss in a well-defined model with experimental cachexia. Indomethacin was provided in the drinking water to explore interactions between host and tumor-derived prostaglandins and proinflammatory cytokines for tumor growth. Wild-type tumor-bearing mice developed cachexia because of rapid tumor growth, which were both attenuated in IL-6 gene knockouts. Similar findings were observed after provision of anti-IL-6 to wild-type tumor-bearing mice. Alterations in food intake were not directly related to systemic IL-6 but rather secondarily to IL-6-dependent tumor growth. The absence of host-derived IL-12, IFN-gamma, or the TNF receptor 1 or receptor 2 gene did not attenuate tumor growth or improve subsequent cachexia. Thus, carcass weight loss was not improved by the omission of host cytokine (TNF-alpha, IL-12, or IFN-gamma) except for IL-6. Systemic indomethacin provision decreased plasma prostaglandin E2 in five of six groups of gene knockout tumor-bearing mice, which was associated with improved carcass weight in these groups. Indomethacin seemed to improve food intake to a similar extent in both wild-type and gene knockouts, which agree with the speculation that eicosanoids are more important to explain anorexia than host cytokines. Our results demonstrate that host- and tumor derived cytokines and prostaglandins interact with tumor growth and promote cachexia in a more complex fashion than usually presented based on previous information in studies on either anti-cytokine experiments in vivo or on gene knockouts with respect to a "single cytokine model." Overall, host cytokines were quantitatively less important than tumor-derived cytokines to explain net tumor growth, which indirectly explains subsequent cachexia and anorexia. PMID- 11034093 TI - Down-Regulation of prostate-specific antigen expression by ligands for peroxisome proliferator-activated receptor gamma in human prostate cancer. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily. Recent studies found that ligand-activated PPARgamma regulated differentiation and clonal growth of several types of cancer cells, including prostate cancer, suggesting that PPARgamma could be a tumor suppressor. Troglitazone was a widely used antidiabetic drug that activates PPARgamma. Recently, we reported that this agent had antiprostate cancer effects in vitro and in vivo. In this study, we administered troglitazone for over 1.5 years to an individual with occult recurrent prostate cancer. Using the prostate specific antigen (PSA) levels as a surrogate marker of the disease, the oral administration of troglitazone (600-800 mg/day) reduced the increase velocity of PSA levels, suggesting clinical efficacy of troglitazone in prostate cancer. PSA promoter/ enhancer reporter assays showed that the PPARgamma ligands troglitazone (10(-5) M), pioglitazone (10(-5) M), or 15-deoxy-delta12,14-prostaglandin J2 (10( 5) M) down-regulated androgen-stimulated reporter gene activity in LNCaP cells, a prostate cancer cell line. The PSA promoter contains androgen receptor response elements (AREs). Reporter gene studies showed that troglitazone inhibited androgen activation of the AREs in the PSA regulatory region. Consistent with inhibition of gene expression, 2 days of incubation of LNCaP with troglitazone dramatically suppressed PSA protein expression without suppressing AR expression, suggesting that troglitazone inhibited ARE activation by a mechanism other than down-regulation of expression of the AR. Taken together, ligands of PPARgamma may be a useful therapeutic approach for the treatment of prostate cancer and may be acting, in part, by inhibiting transactivation of androgen-responsive genes. PMID- 11034094 TI - Tk, a new colon tumor-associated antigen resulting from altered O-glycosylation. AB - Erythrocyte polyagglutination antigens T and Tn are truncated O-glycan chains that are also carcinoma-associated antigens. We investigated whether Tk polyagglutination antigen could similarly be a carcinoma-associated marker and a target of immunotherapy. Monoclonal antibody LM389 was raised against Tk erythrocytes and tested by immunohistochemistry. LM389 strongly reacted with 48% human colorectal carcinomas. Labeling of normal tissues was visible on epithelial cells, mainly digestive, but was confined at a supranuclear level. Expression of the antigen on cloned human carcinoma cells correlated with sialosyl-Tn expression. O-Sialoglycoprotein endopeptidase treatment revealed that on carcinomas and cell lines, the epitope was present on O-glycans. Antibody specificity was determined using synthetic carbohydrates. Direct binding and inhibition studies indicated that LM389 best ligands were terminated by two branched N-acetylglucosamine units. Screening of murine cellular cell lines with LM389 allowed development of an experimental model with Tk-positive and -negative cells in syngeneic BDIX rats. Vaccination of rats with Tk erythrocytes provided a protection against growth of rat Tk-positive, but not of Tk-negative, tumor cells in association with the development of antibodies. Taken together, the results indicate that Tk polyagglutination antigen is a new colorectal carcinoma associated antigen, absent from the normal cell surface, resulting from alteration of O-glycans biosynthesis and with potential as a target of immunotherapy. PMID- 11034095 TI - High steady-state levels of p53 are not a prerequisite for tumor eradication by wild-type p53-specific cytotoxic T lymphocytes. AB - CTLs specific to p53 were previously shown to efficiently eradicate p53 overexpressing tumor cells in vitro as well as in vivo. In this report, we demonstrate that these CTLs can also eliminate tumors that display moderate or even low levels of p53. Neither high steady-state levels of p53 nor elevated p53 synthesis is a prerequisite for recognition of tumors by p53-specific CTLs. Instead, our data show that a high p53 turnover rate is an important factor in determining the sensitivity of tumor cells to p53-specific CTLs. Our data suggest that p53 turnover is related to the MHC class I-restricted presentation of p53 derived epitopes at the tumor cell surface and indicate that CTL-mediated immunotherapy that targets p53 can be applied to a wider range of tumors than has thus far been anticipated. PMID- 11034096 TI - Therapeutic efficacy of OX-40 receptor antibody depends on tumor immunogenicity and anatomic site of tumor growth. AB - The OX-40 receptor (OX-40R) is a cell surface glycoprotein of the tumor necrosis factor receptor family that is expressed primarily on activated CD4 T cells. Engagement of OX-40R by the OX-40 ligand (OX-40L) is known to costimulate the production of cytokines by activated T lymphocytes and to rescue effector T cells from activation-induced cell death. It was previously reported that in vivo ligation of OX-40R by administration of OX-40L:immunoglobulin fusion protein or OX-40R monoclonal antibody (mAb) resulted in a significant prolongation of survival of tumor-bearing mice in four histologically distinct solid tumors. In this study, we demonstrate that the therapeutic efficacy of OX-40R mAb was influenced by the tumor burden, the intrinsic immunogenicity of the tumor as well as by the histological site of tumor growth. Whereas subdermal and intracranial growth of weakly immunogenic MCA 203 and MCA 205 sarcomas and GL261 glioma were susceptible to the mAb treatment, established pulmonary MCA 205 metastases were refractory to the same regimen of treatment. Furthermore, the mAb administration had no impact on the growth of the poorly immunogenic B16/D5 mela noma. Tumor regression mediated by OX-40R mAb was dependent on the participation of both CD4 and CD8 T cells and as a result of tumor rejection, a long-term tumor-specific immunity was established. Analysis of tumor-infiltrating T cells revealed the presence of a far greater number of OX-40R+ T cells of both CD4 and CD8 phenotypes in the intracranial immunogenic GL261 glioma than that in the poorly immunogenic B16/D5 melanoma. These results suggest that ligation of OX-40R on activated T cells in situ in the tumor may provide a necessary costimulatory signal to augment immune responses leading to tumor regression and immunological memory. PMID- 11034097 TI - Prostate stem cell antigen is a promising candidate for immunotherapy of advanced prostate cancer. AB - Immunotherapy of prostate cancer (CaP) may be a promising novel treatment option for the management of advanced CaP. However, the lack of suitable tumor antigens remains a major obstacle for the rational design of vaccines. To characterize potential CaP antigens, we determined the mRNA expression of the prostate specific genes C1, C2, C5, PAGE-1, and prostate stem cell antigen (PSCA) in hormone-refractory CaP, benign prostatic hyperplasia, CaP cell lines, and CaP specimens. Among these gene products, only expression of PSCA appears to be retained in the majority of advanced CaP samples, as shown by reverse transcription-PCR analyses. Peptide fragments of PSCA presented in the context of major histocompatibility molecules could serve as recognition targets for CD8 T cells, provided these lymphocytes were not clonally deleted or peripherally tolerized. Our goal was to determine whether the human T-cell repertoire could recognize PSCA-derived peptide epitopes in the context of a common class I allele, HLA-A0201. Of nine peptides that, according to HLA-A0201 binding motifs, were candidate ligands of A0201 class I molecules, three peptides were able to stabilize HLA-A0201 molecules on the cell surface. One of the latter peptides, encompassing amino acid residues 14-22, was capable of generating a PSCA-specific T-cell response in a human lymphocyte culture from a patient with metastatic CaP. PSCA-specific CTLs recognized peptide-pulsed targets as well as three prostate carcinoma lines in cytotoxicity assays, indicating that this peptide could be endogenously processed. In conclusion, our findings establish PSCA as a potential target for antigen-specific, T cell-based immunotherapy of prostate carcinoma. PMID- 11034098 TI - A putative oncogenic role for MPP11 in head and neck squamous cell cancer. AB - Genetic alterations of chromosome 7 are common in human cancer. Furthermore, previous studies have supported the presence of a gene important in a broad range of cancers at 7q22-31.1. There is evidence that supports an oncogenic function for this putative gene, as well as evidence that supports a tumor suppressive role. In this study, we used a cross-species candidate gene approach in combination with physical mapping to identify MPP11 as a candidate for the putative cancer-related activity at 7q22-31.1. We then analyzed primary head and neck squamous cell tumors (HNSCCs) for loss of heterozygosity/allelic imbalance (LOH/AI) at the MPP11 genomic locus. Thirty-eight percent of tumors examined displayed LOH/AI involving the MPP11 genomic locus. Mutation analysis of MPP11 in the latter samples did not identify any inactivating mutations. However, immunohistochemical staining of primary tumor sections and Western blot analysis of HNSCC cell lines revealed a tumor-specific high level of expression of MPP11p. Fluorescence in situ hybridization analysis done on the cell lines identified increased chromosome 7 copy number with a concomitant increase in MPP11 copy number. These results suggest an oncogenic role for MPP11 in HNSCC. PMID- 11034099 TI - Survey of genetic alterations in gastrinomas. AB - Gastrinomas are rare gastrin-secreting endocrine tumors that usually arise in the duodenum or pancreas and, if untreated, can cause severe peptic ulcers or metastatic disease. Although most tumors are sporadic they are especially common in patients with multiple endocrine neoplasia type 1 (MEN1), and most studies of these tumors have focused on the role of the MEN1 gene. Although the gene is commonly altered in sporadic tumors, this finding is not universal, and it is highly likely that other genetic defects play a significant role. In the present study, an in-depth analysis of the DNA of eight tumors was carried out in an effort to localize these areas. The experiments consisted of an analysis of 400 microsatellite marker loci distributed evenly throughout the human genome, and the results were confirmed with comparative genomic hybridization. Whereas deletions encompassing the MEN1 gene were seen in two tumors, the most striking result was multiple large rearrangements on chromosome 1 in two of the tumors with hepatic metastases. In several instances, an individual tumor had abnormalities of every informative maker on a given chromosome, presumably as a result of aneuploidy affecting that chromosome. Such defects were only seen in the four large or aggressive tumors, and the total number of chromosomes affected in a tumor ranged from 1 to a high of 13 in a patient who had an unusually aggressive tumor This tumor also showed microsatellite instability, and this is the first report of such a defect in gastrinomas. This study implicates chromosome 1 defects, aneuploidy, and perhaps mismatch repair defects as importan features of gastrinomas; deletions involving the MEN1 gene were con firmed, but the rest of the genome was free of large deletions or amplifications. PMID- 11034100 TI - Defects in methylthioadenosine phosphorylase are associated with but not responsible for methionine-dependent tumor cell growth. AB - A large proportion of human tumor-derived cell lines and primary tumor cells show methionine-dependent growth. This phenomenon refers to the ability of cells to grow in media containing methionine and the inability of cells to grow in media supplemented with methionine's precursor, homocysteine (Hcy). Methionine can be formed by two different pathways, the recycling pathway and the salvage pathway. To discover the basis for methionine-dependent growth, we have analyzed 12 tumor cell lines and 2 non-tumor-derived cell lines for defects in two key genes in different methionine synthetic pathways. We found little evidence that defects in methionine synthase expression or mutations in the MS gene are correlated with methionine-dependent growth. However, we did find a correlation between methionine-dependent growth and defects in expression of methylthioadenosine phosphorylase (MTAP), a key enzyme in the salvage pathway. Three of the four cell lines lacking detectable MTAP protein were unable to grow in Hcy-containing media, whereas all six of the MTAP-positive cell lines tested showed strong growth. However, when we introduced MTAP cDNA into MTAP-deficient MCF-7 cells, the resulting cell line was still defective in growth on Hcy, although it could now grow on the salvage pathway precursor methylthioadenosine. These findings indicate that salvage pathway defects are not causally related to methionine dependent growth. PMID- 11034101 TI - BRCA1 and BRCA2 are necessary for the transcription-coupled repair of the oxidative 8-oxoguanine lesion in human cells. AB - The breast and ovarian cancer susceptibility genes, BRCA1 and BRCA2, are likely to participate in DNA lesion processing. Oxidative lesions, such as 8-oxoguanine, occur in DNA after endogenous or exogenous oxidative stress. We show that deficiency for either BRCA1 or BRCA2 in human cancer cells leads to a block of the RNA polymerase II transcription machinery at the 8-oxoguanine site and impairs the transcription-coupled repair of the lesion, leading to a high mutation rate. Expression of wild-type BRCA1 from a recombinant adenovirus fully complements the repair defect in BRCA1-deficient cells. These results represent the first demonstration of the essential contribution of BRCA1 and BRCA2 gene products in the repair of the 8-oxoguanine oxidative damage specifically located on the transcribed strand in human cells. This suggests that cells from individuals predisposed to breast and/or ovarian cancer may undergo a high rate of mutations because of the deficiency of this damage repair pathway after oxidative stress. PMID- 11034102 TI - Screening of the Men1 gene and discovery of germ-line and somatic mutations in apparently sporadic parathyroid tumors. AB - Hyperparathyroidism is the first manifestation in a majority of multiple endocrine neoplasia (MEN1) patients. To discriminate between sporadic and hereditary parathyroid tumors and characterize MEN1 somatic mutations, we examined MEN1 gene mutations in patients who had undergone surgery for sporadic parathyroid tumors. DNA was extracted from fresh frozen parathyroid tumor specimens from 112 patients as well as from peripheral blood leukocytes from 64 of the 112 patients. Sequence analysis was performed to examine exons 2-10 of the MEN1 gene for mutations. Loss of heterozygosity (LOH) was also examined by an analysis of codon 418 and 541, which lie within a polymorphic region of MEN1. Somatic MEN1 mutations were found in 25 of the 112 patients (22%). Two patients had two point mutations (508del33 and Y341X and 363insT and 1767delT, respectively). A total of 27 mutations were characterized, 20 of which have not been reported previously. There were 7 nonsense mutations, 10 frameshift mutations, 2 splice site deletions, 5 missense mutations, and 3 in-frame mutations. Nineteen mutations (70%) predicted truncation of the menin protein. Germ-line MEN1 mutations were found in 3 of 64 patients (5%) who had no family history of endocrine tumors associated with MEN1, and these patients were identified as MEN1 gene probands. LOH at the MEN1 locus was detected in three parathyroid tumors showing germ-line mutation. LOH was significantly frequent in parathyroid tumors with somatic MEN1 mutations (15 of 22 tumors, 68%) but not in those without germ-line or somatic MEN1 mutations (14 of 51 tumors, 28%; P = 0.0011). Our findings suggest that alterations of both alleles of the MEN1 gene may be associated not only with endocrine tumors of affected MEN1 patients but also with sporadic parathyroid tumors. Germ-line MEN1 gene analysis can distinguish heritable from nonheritable parathyroid tumors, and MEN1 gene evaluation of patients with apparently sporadic parathyroid tumor is recommended before parathyroid surgery. PMID- 11034103 TI - Activation of peroxisome proliferator-activated receptor gamma by troglitazone inhibits cell growth through the increase of p27KiP1 in human. Pancreatic carcinoma cells. AB - In the present study, we examine whether human pancreatic carcinoma cells express peroxisome proliferator-activated receptor gamma (PPARgamma) and the effect of PPARgamma activation by its selective ligand on cellular growth in pancreatic cancer cells. Immunohistochemical study of resected human pancreata using a polyclonal PPARgamma antibody revealed that PPARgamma protein expression in the nuclei of carcinoma cells was observed in 9 of 10 pancreatic adenocarcinomas. In contrast, normal pancreatic duct epithelial cells in the samples expressed no PPARgamma. Reverse transcription-PCR and Northern blot analysis demonstrated that all four tested human pancreatic cancer cell lines, PK-1, PK-8, PK-9, and MIA Paca-2, expressed PPARgamma mRNA. Luciferase assay in PK-1 cells showed that troglitazone, a selective ligand for PPARgamma, transactivated the transcription of a peroxisome proliferator response element-driven promoter in a dose-dependent fashion. Troglitazone inhibited the growth of all four pancreatic carcinoma cell lines in a dose-dependent manner. Cell cycle analysis by flow cytometry demonstrated that troglitazone induced G1 arrest in PK-1 cells. To examine the role of cyclin-dependent kinase inhibitors in the G1 arrest by troglitazone, we determined p27KiP1, p21CiP1/Waf1, or p18Ink4c protein expression by Western blot analysis in troglitazone-treated PK-1 cells. Troglitazone increased p27Kip1 but not p21Cip1/Waf1 or p18Inkc protein levels in time- and dose-dependent manners. To clarify the functional importance of p27Kip1 in the cell growth inhibition by troglitazone. All these results suggest that PPARgamma could be considered as a possible target molecule for treatment in human pancreatic carcinomas. PMID- 11034104 TI - Anti-Vascular endothelial growth factor treatment augments tumor radiation response under normoxic or hypoxic conditions. AB - Recent studies in experimental animals have shown that combining antiangiogenic therapy with radiation can enhance tumor response. Whether this enhancement is mainly attributable to angiogenesis inhibition, endothelial cell radiosensitivity, tumor cell apoptosis, or a decrease in the number of hypoxic cells (improved oxygenation) is not known. We designed this study to discern the role of tumor oxygenation. We chose an anti-vascular endothelial growth factor (anti-VEGF) monoclonal antibody (mAb) which has a known target, human VEGF. We also measured interstitial fluid pressure (IFP) to test the hypothesis that the decreased vascular permeability induced by the anti-VEGF mAb can lower IFP. The effect of anti-VEGF mAb on vascular density, partial oxygen tension (pO2), and apoptosis was also measured. Athymic NCr/Sed nu/nu mice bearing 6-mm xenograft of the human glioblastoma multiforme (U87), or colon adenocarcinoma (LS174T) were treated with anti-VEGF mAb injected i.p. on alternate days for a total of six injections at a dosage of 100 microg/injection/mouse. For combined anti-VEGF and radiation, single radiation doses were given under normal blood flow (20 and 30 Gy) or clamped hypoxic conditions (30 and 40 Gy) 24 h after the sixth injection of mAb. The inhibition of the growth of U87 and LS174T tumors by the anti-VEGF mAb was associated with a significant reduction in tumor vascular density and a relatively small increase in the number of apoptotic cells. Compared with size matched controls, IFP decreased by 74% in LS174T, and 73% in U87 in mice treated with anti-VEGF mAb. After antibody treatment PO2 increased significantly in U87, but did not change in LS174T tumors. Combined treatment induced in U87 tumors a tumor-growth delay (TGD) which was greater than additive; in LS174T except for the 40-Gy hypoxic group, the effect was only additive. In both U87 and LS174T the TGD induced by the antibody was independent of oxygen levels in the tumor at the time of radiation. The fact that the increase in TGD occurred under both normoxic and hypoxic conditions suggests that anti-VEGF mAb treatment can compensate for the resistance to radiation induced by hypoxia. PMID- 11034105 TI - The caM kinase, Pnck, is spatially and temporally regulated during murine mammary gland development and may identify an epithelial cell subtype involved in breast cancer. AB - While screening for protein kinases expressed in the murine mammary gland, we identified previously a Ca2+/calmodulin-dependent kinase, Pnck, that is most closely related to CaMKI. In this report, we show that Pnck is temporally regulated during murine mammary development with highest levels of expression observed late in pregnancy, concomitant with the decreased cellular proliferation and terminal differentiation of the mammary epithelium. Consistent with this finding, Pnck is up-regulated in confluent mammary epithelial cells and is down regulated as serum-starved cells are stimulated to reenter the cell cycle. In the mammary gland, Pnck is expressed in an epithelial-specific and markedly heterogeneous manner, suggesting that the expression of this kinase may be restricted to a particular mammary epithelial cell type. Potentially related to its heterogeneous in vivo expression pattern, Pnck expression is oncogene associated in murine epithelial cell lines derived from mammary tumors arising in different transgenic mouse models of breast cancer; cell lines derived from mammary tumors initiated by c-myc or int-2/Fgf3 express Pnck, whereas cell lines initiated by neu or H-ras do not. In an analogous manner, expression of the human homologue of Pnck is restricted to a subset of human breast cancer cell lines. Moreover, PNCK was found to be highly overexpressed in a subset of human primary human breast cancers compared with benign mammary tissue. Together, our data suggest that Pnck may play a role in mammary development, and that expression of this kinase may be restricted to a mammary epithelial cell type that is transformed in a subset of human breast cancers. PMID- 11034107 TI - Expression of BARF1 gene encoded by Epstein-Barr virus in nasopharyngeal carcinoma biopsies. AB - We reported previously that the EBV BARF1 open reading frame encodes a Mr 31,000 33,000 protein (p31) with potential transforming and oncogenic properties. This gene was found capable of transforming both: (a) the rodent fibroblast lines Balbc/3T3 and NIH3T3 into cells producing aggressive tumors in newborn rats; and (b) the human EBV-negative B-cell line Louckes into cells leading to small tumors, which disappeared 3 weeks after injection. Our recent study showed that BARF1 ORF expression may confer the property of immortalization to primary kidney epithelial cells (M. X. Wei et al., Oncogene, 14: 3073-3081, 1997). Because this suggested that BARF1 could be involved in epithelial malignancy, we investigated its transcriptional and translational expressions in Algerian nasopharyngeal carcinoma (NPC) biopsies by reverse transcription-PCR and immunoblotting using rabbit polyclonal antisera prepared against two synthetic peptides corresponding to distinct, predicted epitopes of the BARF1 protein (NGGVMKEKD, amino acids 172 180, and GKNDKEE, amino acids 203-209). The BARF1 ORF was found to be transcribed and translated in >85% of our NPC biopsies, with high p31 protein level detected in several NPC patient biopsies as well as in NPC-derived xenografts. Our observation of BARF1 expression in a large proportion of NPC epithelial cells suggests that this EBV gene might play an important role in the malignant transformation of human epithelial cells in vivo. PMID- 11034106 TI - Expression of a novel factor, com1, is regulated by 1,25-dihydroxyvitamin D3 in breast cancer cells. AB - Tumor cells and their surrounding microenvironment produce a variety of factors that promote tumor growth and metastasis. We recently identified a nuclear factor, termed com1, that is up-regulated in human breast carcinoma cells on formation of experimental metastatic tumors and is assumed to act as a growth promoting factor in breast cancer. 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] is a potent inhibitor of growth in breast cancer both in vitro and in vivo. We compared the growth-regulatory mechanisms of nontumorigenic and estrogen dependent MCF-7 cells with those of the tumorigenic and tamoxifen-resistant subline MCF7/ LCC2 in the presence of 1,25(OH)2D3. Proliferation of MCF7/LCC2 cells, which revealed constitutive com1 expression, was inhibited by 1,25(OH)2D3 (10(-7) M). This was strongly associated with cell cycle arrest in G1 phase, consistent with accumulation of the hypophosphorylated form of the retinoblastoma protein as well as the induction of the cyclin-dependent kinase inhibitor p21. These cell cycle events were preceded by a transient up-regulation (5-8-fold) of com1 mRNA. Furthermore, clonal growth of the MCF7/LCC2 cells was also inhibited by 1,25(OH)2D3 (10(-7) M), and when the com1-negative MCF-7 cells were stably transfected with com1, the resulting MCF7/com1 cells showed a significant decrease in colony formation. These results seem to indicate that rather than promoting growth, com1 may participate in the regulatory pathway involved in cellular growth inhibition when recruited by inhibitory signals. PMID- 11034108 TI - Differences in the immunogenicity of latent membrane protein 1 (LMP1) encoded by Epstein-Barr virus genomes derived from LMP1-positive and -negative nasopharyngeal carcinoma. AB - We have previously shown that an EBV-encoded latent membrane protein 1 (LMP1) gene derived from a nude mouse-propagated nasopharyngeal carcinoma (NPC) tumor and expressed in nonimmunogenic murine mammary carcinoma S6C cells failed to convey immunogenicity (rejectability) in syngeneic mice, whereas the corresponding B-cell derived LMP1 gene made the mice highly immunogenic. This raised the question of whether LMPL-expressing NPCs have been selected for low immunogenicity at the viral gene expression level. If so, LMP1-negative tumors that carry highly methylated LMP1 regulatory sequences may not have been exposed to a similar immunoselection. In the present study, we have compared LMP1 genes derived from two LMP1-positive NPCs and two LMP1-negative NPCs. All four genes were expressed in S6C cells in parallel with the previously tested isolates from a B-cell (B95-8)-derived and a nude mouse-propagated NPC (Cao)-derived gene. As in the previous study, we have found that the B-cell-derived LMP1 isolate was highly immunogenic. LMP1-positive tumor-derived isolates were poorly immunogenic, whereas the isolates from the LMP1-negative NPC tumor had intermediate immunogenicity. Sequence data revealed that LMP1 genes from LMP1-expressing NPC had 16 amino acid substitutions, whereas LMP1 from non-LMP1-expressing NPC had only 9 amino acid changes in the coding region. Three of the changes were at shared sites, but with different modifications. The fact that the gene from non LMP1-expressing NPC mutated at a low frequency but was more immunogenic than the LMP1 gene derived from LMP1-expressing NPC, which was highly mutated but less immunogenic, favors the idea that LMP1-positive tumors escape immunosurveillance in immunocompetent hosts by either a selective down-regulation of LMP1 expression, methylation in the LMP1 promoter sequence, or mutation of LMP1 in LMP1-expressing samples. PMID- 11034109 TI - Correlation between nuclear melatonin receptor expression and enhanced cytokine production in human lymphocytic and monocytic cell lines. AB - The report shows that melatonin enhances IL-2 and IL-6 production by two human lymphocytic (Jurkat) and monocytic (U937) cell lines via a nuclear receptor mediated mechanism. Jurkat cells express nuclear (RZRalpha, RORalpha1 and RORalpha2) and membrane (mt1) melatonin receptors, and melatonin binds to Jurkat nuclei and membranes with the same affinity described for human peripheral blood mononuclear cells (PBMCs). Melatonin enhances IL-2 production by Jurkat cells activated by either phytohemagglutinin (PHA) or phorbol myristate acetate (PMA). PHA activation of Jurkat cells does not change the profile of melatonin receptor expression; on the contrary, PMA activation negatively regulates the mtl receptor. In the absence of the membrane receptor, melatonin still activates IL-2 production. U937 cells express only the mtl receptor. Although melatonin binds to both U937 nuclei and membranes, CGP 52608, a ligand of the nuclear receptor for melatonin, does not inhibit melatonin binding to U937 nuclei, suggesting that a protein other than the RZR/RORalpha receptor was involved in the process. In U937 cells, melatonin did not modify basal production of IL-6 or when activated by PMA plus LPS (lipopolysaccharide), a treatment that downregulates the expression of the mtl receptor. However, in U937 cells activated with IFN-gamma, which induces the expression of the RORgamma1 and RORalpha2 nuclear receptors and represses the expression of the mt1 receptor, melatonin can activate IL-6 production. These results show that the expression of nuclear melatonin receptor is sufficient for melatonin to activate cytokine production in human lymphocytic and monocytic cell lines. PMID- 11034110 TI - Low urinary 6-sulphatoxymelatonin levels in patients with coronary artery disease. AB - A decrease in nocturnal serum melatonin levels was reported in patients with clinically uncharacterized coronary artery disease. To assess whether there was a correlation between melatonin production and disease stage, we measured the nocturnal urinary excretion of 6-sulphatoxymelatonin (an index of blood melatonin concentration) in patients with chronic stable or unstable coronary disease and in a group of age-matched controls. Three groups of individuals were studied: a) 24 healthy subjects (mean age: 63 +/- 13 yr); b) 32 patients with chronic, stable, coronary disease (62 +/- 11 yr); and c) 27 patients with unstable angina (62 +/- 12 yr). For 6-sulphatoxymelatonin measurement, urine was collected from 18:00 to 06:00 hr, within 48 hr of hospitalization in the case of unstable angina. 6-Sulphatoxymelatonin was measured by a specific radioimmunoassay. Urinary 6-sulphatoxymelatonin excretion was significantly lower in unstable angina patients than in healthy subjects or in patients with stable angina. 6 Sulphatoxymelatonin correlated negatively with age in healthy subjects, but not in coronary patients. 6-Sulphatoxymelatonin excretion in patients treated with beta-adrenoceptor blockers did not differ significantly from coronary patients not receiving beta-blockers. The results indicate that patients with coronary disease have a low melatonin production rate, with greater decreases in those with higher risk of cardiac infarction and/or death. PMID- 11034111 TI - Melatonin protects against stress-induced gastric lesions by scavenging the hydroxyl radical. AB - The antiulcer effect of melatonin on gastric lesions caused by restraint-cold stress or by indomethacin (IMN) was studied with the intent of determining the mechanism of action of the indole. Melatonin dose-dependently prevents both stress and IMN-induced gastric damage with around 90% inhibition at a dose of 60 mg per kg BW. When compared with already-marketed antiulcer drugs, such as ranitidine and omeprazole, melatonin was found to be more effective than ranitidine but less effective than omeprazole in preventing stress ulcer. When compared with other antioxidants, melatonin was more potent than glutathione and essentially equipotent to alpha-tocopherol in blocking stress-induced ulcer. As stress-induced gastric lesions are mainly caused by oxidative damage due to hydroxyl radicals (*OH), the effect of melatonin in scavenging the *OH generated during stress conditions, as well as in an in vitro model system, was studied. The results indicate that melatonin at the dose of 60 mg per kg BW caused an 88% reduction of endogenous *OH during stress. Melatonin was also highly effective in scavenging *OH generated in vitro by a Cu2+-ascorbate system. In this case, melatonin at 100 microM reduced *OH by 80%. Melatonin was also found to be a more potent radical scavenger than benzoate, a known *OH scavenger. The results indicate that melatonin prevents stress-induced gastric lesions by scavenging the endogenous *OH. As it also protects against IMN-induced gastric damage, it probably also offers gastroprotection by maintaining endogenous prostaglandin levels. PMID- 11034112 TI - Different circulatory response to melatonin in postmenopausal women without and with hormone replacement therapy. AB - In young men and women, melatonin influences vascular reactivity and reduces blood pressure and norepinephrine levels. Herein, we investigated whether these effects are conserved in postmenopausal women without and with hormone replacement therapy (HRT). Oral melatonin (1 mg) or placebo was randomly and in double blind fashion administered to 18 untreated and 13 postmenopausal women who were treated continuously with transdermal estradiol (50 microg/day) plus cyclic medroxyprogesterone acetate (5 mg/day x 12 days every 28 days). Internal carotid artery pulsatility index (PI), an index of downstream resistance to blood flow, blood pressure and catecholamine levels were evaluated. In untreated postmenopausal women, melatonin was ineffective, while in HRT-treated women, studied during the only estrogenic phase, melatonin reduced, within 90 min, systolic (-8.1 +/- 9.9 mmHg; P = 0.054), diastolic (-5.0 +/- 7.0 mmHg; P = 0.049) and mean (- 6.0 +/- 6.6 mmHg; P = 0.037) blood pressure. Norepinephrine (-50.1 +/ 66.7 pg/mL; P = 0.019), but not epinephrine levels, were also significantly reduced. Similarly, resistance to blood flow in the internal carotid artery, as evaluated by the PI, decreased (-0.190 +/- 0.15; P = 0.0006) in a way that was linearly related to pre-existing PI values (r2 = 0.5; P = 0.0059). These data show that the circulatory response to melatonin is conserved in postmenopausal women on HRT but not in untreated postmenopausal women. Possible physiological and pharmacological implications of these data on the cardiovascular risk of postmenopausal women can be envisioned. PMID- 11034113 TI - Attenuation by melatonin of human umbilical arterial vasoconstriction induced by lysophosphatidylcholine. AB - We evaluated the antioxidant property of melatonin as related to the vasospastic effect of lysophosphatidylcholine (LPC), a component of oxidized lipoprotein, on the human umbilical artery. Helical sections of umbilical arteries were obtained from healthy pregnant women who were delivered between 37 and 39 wk of gestation. Changes in maximal tension induced by KCl were measured in arterial sections having intact endothelium. Sections were treated with LPC alone (15 or 30 microM), or were pretreated either with a hydrogen peroxide (H2O2) scavenger (catalase, 1,200 U/mL), a hydroxyl radical scavenger (mannitol, 30 mM), a nitric oxide (NO) synthesis inhibitor (L-N(G)-monomethyl arginine, LNMA, 2 x 10(-4) M) or melatonin (1 or 10 microM). The effect of LPC (30 microM) on the vasorelaxation induced by 5-hydroxytryptamine (5-HT) was also determined, with or without melatonin pretreatment (10 microM). LPC potentiated vascular tension in a concentration-dependent manner. Pretreatment with LNMA significantly suppressed this vasospastic effect of LPC. Pretreatment with catalase or mannitol significantly reduced the vasospastic effect of LPC. Melatonin significantly lessened the vasospastic effect of LPC in a concentration-dependent manner. Pretreatment with LPC significantly inhibited the relaxation induced by 5-HT. Treatment with melatonin prior to LPC exposure significantly restored the relaxation induced by 5-HT. Results suggest that LPC potentiates vascular tension in human umbilical artery, perhaps by suppressing the endothelial synthesis of NO. Melatonin significantly suppressed the vasospastic effect of LPC. This agent probably scavenges the hydroxyl radicals arising from LPC. PMID- 11034114 TI - Melatonin effects on intercellular junctional communication in MCF-7 human breast cancer cells. AB - Melatonin exerts a direct antiproliferative effect on estrogen-responsive MCF-7 cells in culture. Recently, the importance of the anti-invasive actions of melatonin as a part of the oncostatic action of this indolamine has been reported. Gap junctional intercellular communication is known to be involved in controlling cell proliferation and differentiation, and a decrease in intercellular junctional communication has been described in highly invasive mammary cancer cells. Because melatonin at physiological doses (1 nM) shifts MCF 7 cells to a lower invasive status, we postulate that melatonin could modulate the levels of gap junctional intercellular communication in these tumor cells. To test our hypothesis, we studied gap junctional intercellular communication in MCF 7 human breast cancer cells previously (7-8 days) treated, or not, with melatonin (10 microM or 1 nM). Using the scrape-loading assay dye-transfer technique to introduce 0.05% Lucifer yellow into cells, we measured the ability of the tumor cells to transfer dye to adjacent cells. Rhodamine dextran (0.05%) was used as a control dye to verify that dye-transfer occurs through intercellular junctions. The presence of melatonin (10 microM or 1 nM) in the culture medium significantly increased (P < 0.01) the transfer of the dye to adjacent cells through gap junctions. This increase was greater at 10 microM melatonin, and averaged scan profiles of cells treated with melatonin 10 microM showed a statistically significant increase (P < 0.01) in the integrated optical density values, and a broadening of the densitometric scan. These findings suggest that melatonin could exert its antitumor action, at least in part, by increasing regulatory signals that are passed between adjacent epithelial cells through intercellular junctions. PMID- 11034115 TI - Potential involvement of mt1 receptor and attenuated sex steroid-induced calcium influx in the direct anti-proliferative action of melatonin on androgen responsive LNCaP human prostate cancer cells. AB - Melatonin, a pineal secretory product, has been shown to exert a direct anti proliferative action on the androgen-sensitive LNCaP prostate cancer cell line through hitherto undefined mechanisms. In this communication, expression of mt1 melatonin receptor protein in human prostate cancer tissues and LNCaP cells was demonstrated by immunohisto(cyto)chemistry and western blotting, hence supporting the use of LNCaP cell line as a model for the study of melatonin signaling in prostate cancer cell growth. Using 3H-thymidine incorporation assay, LNCaP cell proliferation was inhibited by 2-iodomelatonin, a high-affinity melatonin receptor agonist. Furthermore, melatonin inhibited 3H-thymidine incorporation into LNCaP cells and attenuated 5alpha-dihydrotestosterone (DHT) or 17beta estradiol (E2)-induced stimulation of LNCaP cell proliferation at physiological and pharmacological concentrations. Similar concentration-dependent inhibition of sex steroid-induced stimulation of thymidine incorporation into LNCaP cells by 2 iodomelatonin was also observed. Interestingly, attenuation of sex steroid stimulated calcium influx into LNCaP cells by pharmacological concentrations of melatonin was recorded, whereas 2-iodomelatonin had no effect on cytosolic calcium changes induced by sex steroids. In addition, proliferative and cytosolic calcium changes were associated with inhibition of total prostate-specific antigen (PSA) production by LNCaP cells at high physiological and pharmacological concentrations of melatonin. Our data suggest that activated mt1 receptor and attenuated sex steroid-induced calcium influx are two important mechanisms mediating the direct anti-proliferative action of melatonin on androgen responsive human prostate cancer cells. PMID- 11034116 TI - Nitrosation of melatonin by nitric oxide and peroxynitrite. AB - Peroxynitrite (ONOO-) is an endogenous molecule, formed by rapid coupling between *NO and O2*-. ONOO- is known to be a strong oxidant of thiols and metalloorganic compounds and also a nitrating agent of aromatic compounds such as tyrosine. However, its chemistry is not yet well elucidated under physiological conditions. Melatonin, which is an indole-amine produced by the pineal gland and other organs, has antioxidant properties. We show that melatonin reacts with ONOO- in phosphate-buffered solutions. We provide evidence of nitrosation and oxidation at the pyrrole nitrogen leading to 1-nitrosomelatonin and 1-hydroxymelatonin, these being the major reactions in aqueous phosphate-buffered solutions besides other aromatic hydroxylations and nitration. 4-Nitromelatonin is formed, but in small amounts. The kinetics of all transformations were strictly dependent on ONOO- decay, whereas yields varied with pH and the presence of CO2. The N-oxidation became competitive with nitrosation at pH 7.4, in medium containing a sufficient amount of CO2. A proposed mechanism involves the transient formation of melatonyl radical and ONOO* radical derived from ONOO- decay. PMID- 11034117 TI - Treatment of dentine hypersensitivity by lasers: a review. AB - BACKGROUND, AIMS: Since the development of the ruby laser by Maiman in 1960, a variety of papers on potential applications for lasers in dentistry have been published. The purpose of this paper is to summarise laser applications for the treatment of dentine hypersensitivity. The effects of laser on pulp tissue and problems on laser treatment are also reviewed. This article reviews the role of lasers for the treatment of dentine hypersensitivity since 1985, summarises many research reports from the last decade, and surmises what the future may hold for lasers in this treatment. METHOD: To date, 4 kinds of lasers have been used for the treatment of dentine hypersensitivity, and the effectiveness ranged from 5.2 to 100%, which was dependent on the laser type and parameters used. The mechanism involved in laser treatment of dentine hypersensitivity are relatively unknown. RESULTS: These require clarification to result in safely effective treatment optimization. In general, the efficiency for the treatment of dentine hypersensitivity using lasers is higher than other methods, but in severe cases, it is less effective. CONCLUSION: It is necessary to consider the severity of dentine hypersensitivity before laser use. PMID- 11034118 TI - Microbial composition of supra- and subgingival plaque in subjects with adult periodontitis. AB - BACKGROUND, AIMS: The purpose of the present study was to compare and relate the microbial composition of supra and subgingival plaque in 23 adult periodontitis subjects (mean age 51 +/- 14 years). METHODS: A total of 1,170 samples of supra and subgingival plaque were collected from the mesial aspect of every tooth (up to 28 supra and 28 subgingival samples) from each subject and evaluated for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. The counts (levels), % DNA probe count (proportion) and % of sites colonized (prevalence) of each species in supra and separately in subgingival plaque were computed for each subject. Significance of differences between supra and subgingival plaque for each species was sought using the Wilcoxon signed ranks test and adjusted for multiple comparisons. RESULTS: All 40 taxa were detected in both supra and subgingival plaque. Actinomyces species were the most prevalent taxa in both habitats. 75 to 100% of supra and 62 to 100% of subgingival sites were colonized by at least one of the 5 Actinomyces species. Supragingival samples exhibited significantly higher counts of Actinomyces naeslundii genospecies 1, Actinomyces israelii, Actinomyces odontolyticus, Neisseria mucosa, Streptococcus gordonii, Capnocytophaga ochracea and Capnocytophaga sputigena when compared with mean counts in subgingival samples taken from the same tooth surfaces. Subgingival plaque samples presented significantly higher counts of Prevotella nigrescens, Prevotella intermedia, Bacteroides forsythus and Porphykromonas gingivalis. Subgingival samples exhibited a significantly higher proportion of "red" and "orange complex" species, while supragingival plaque exhibited higher proportions of "green" and "purple" complex species as well as Actinomyces species. Suspected periodontal pathogens could be detected in supragingival plaque from sites where subgingival samples were negative for the same species. CONCLUSIONS: The data indicate that supragingival plaque can harbor putative periodontal pathogens, suggesting a possible role of this environment as a reservoir of such species for the spread or reinfection of subgingival sites. PMID- 11034119 TI - Effect of triclosan on interferon-gamma production and major histocompatibility complex class II expression in human gingival fibroblasts. AB - BACKGROUND, AIMS: The effect of triclosan (2,4,4'-trichloro-2'-hydroxyl-diphenyl ether) on the production of interferon-gamma (IFN-gamma) and the expression of major histocompatibility complex (MHC) class II antigen was studied in human gingival fibroblasts isolated from 4 individuals. METHODS/RESULTS: All cell lines demonstrated high IFN-gamma production in 24-h cultures of human gingival fibroblasts stimulated by phytohemagglutinin (PHA) (5 microg/ml). Human gingival fibroblasts showed a high expression of MHC class II when stimulated with 500 and 1,000 pg/ml rIFN-gamma in 7-day cultures. Treatment of the cells with triclosan (0.5 microg/ml) reduced both IFN-gamma production and MHC class II expression in human gingival fibroblast cultures. Similar inhibitory effects on IFN-gamma production and MHC class II expression were observed when the anti-inflammatory agent dexamethazone (1 microM) was used. CONCLUSION: The present study further supports the view that the agent has an anti-inflammatory effect in addition to its antibacterial capacity. PMID- 11034120 TI - Interleukin-1 and IL-1 receptor antagonist in gingival crevicular fluid. AB - BACKGROUND/AIMS: This study aimed to investigate the cytokine IL-1beta and its receptor antagonist IL-1ra in gingival crevicular fluid (GCF), in patients with adult periodontitis. METHOD: A total of 40 GCF samples were harvested from 10 subjects with moderate to severe adult periodontitis and 10 healthy controls. Subjects were selected from both genders, with all the upper anterior teeth present, and with no relevant systemic illness, pregnancy or recent medication. All subjects were non-smokers and had not received any periodontal therapy within the preceding 3 months. Deep bleeding sites, deep non-bleeding sites and healthy sites were investigated in relation to upper anterior teeth. Clinical measurements were recorded for each site, after obtaining a GCF sample. IL-1beta and IL-1ra were quantified using new commercially available ELISA kits (Quantikine), and could be detected in all samples. RESULTS: The mean concentration for IL-1beta was 0.11 (SD 0.14) pg/microl for bleeding periodontitis sites, 0.04 (0.05) pg/microl for non-bleeding periodontitis sites, and 0.01 (0.03) pg/microl for healthy sites (p<0.001). In contrast, the mean concentration for IL-1ra was 6.99 (9.78) pg/microl for healthy sites, 0.59 (0.44) pg/microl for non-bleeding periodontitis sites, and 0.44 (0.36) pg/microl for bleeding periodontitis sites (p<0.001, except for comparisons between bleeding and non-bleeding periodontitis sites, p>0.05). For healthy sites, a strong inverse relationship was found between IL-1beta and IL-1ra levels in GCE. CONCLUSIONS: The results suggest a strong relationship between the severity of adult periodontitis and the increasing GCF levels of IL-1beta and decreasing levels of IL-1ra. PMID- 11034121 TI - Comparison of a double-textured prototype manual toothbrush with 3 branded products. A professional brushing study. AB - BACKGROUND: The consensus has been that there is no one superior design of manual toothbrush for plaque removal, despite, in some cases, encouraging findings from laboratory studies. The user appears the major variable and may mask differences in brush efficacy. AIMS: The aim of this study was to compare 2 new double filament texture brushes with 3 established brands for plaque removal, under standardised conditions of professional brushing. METHOD: The study was a blind, randomised crossover design, balanced for residual effects. A panel of 15 subjects suspended tooth cleaning for 3 days. On day 4, plaque was scored from 6 sites per tooth before and after a 2-min professional toothbrushing. A washout period of at least 3 days was allowed between study periods. RESULTS: Overall plaque removal was 50% with no significant differences between brushes, a 3% absolute difference in average total mouth plaque separating brushes. However, at upper- and mid-buccal sites, a not significant 8% and a significant 9% difference, respectively, in plaque removal were recorded in favour of one of the prototype brushes, and at the mid-lingual site, there was a non-significant 10% difference in favour of one branded brush. Other sites were cleaned similarly by all brushes except for reduced plaque removal from buccal compared to lingual surfaces and interproximal compared to mid-surface sites. Highly significant subject differences in plaque removal were noted which may be relevant to inherent anatomical difficulties in tooth cleaning for some individuals. Period effects were not significant, supporting the consistency in brushing by the professional brusher. CONCLUSION: The method appeared capable of detecting small benefits of brush design. However, the benefits reported must be taken within the context of an overall lack of difference between brushes. The method could be used to set and record a minimum level of efficacy for toothbrushes. PMID- 11034122 TI - The use of professional brushing to compare 3 toothbrushes for plaque removal from individuals with gingival recession. AB - BACKGROUND: Toothbrush filament configuration now varies considerably, usually with the intention of improving plaque removal overall or from specific sites. AIMS: The aim of the study was to compare 2 branded toothbrushes with a brush designed with a convex filament head arrangement to improve plaque removal from teeth with buccal gingival recession. METHODS: The study was a randomised, 3 treatment, double-blind crossover design balanced for residual effects and involving 18 healthy volunteers. On day 1, subjects suspended toothcleaning and returned to the clinic on day 4. Plaque was scored by area and index before and after 2 min of toothbrushing with the allocated brush. Brushing was performed by a hygienist allocating a timed 15 s to each of the 8 buccal and lingual quadrants. 3 days was allowed between treatment periods. RESULTS: Plaque accumulation differed considerably by site as did plaque removal. Overall plaque removal was 40%. Analysis showed significant subject differences but, no significant period or treatment differences. CONCLUSIONS: Consistent with other reports, it must be concluded that brush design is not a major variable in plaque removal. However using this "robot"-like model, the dentition of some individuals and specific dental surfaces are inherently more difficult to clean than others. PMID- 11034123 TI - Effects of hormonal contraceptives on the periodontium, in a population of rural Sri-Lankan women. AB - AIMS: Women using hormonal contraceptives can be considered to be a 'risk group' for periodontal disease, due to prolonged, sustained serum levels of oestrogens and progesterone. This investigation aims to study the effects of hormonal contraceptives on periodontal tissues. METHODS: 32 women using hormonal contraceptives for less than 2 years, 17 for 2-4 years and a matched control group of 39 non-users were selected for the study. They were clinically examined for plaque levels (plaque index: PLI), gingival condition (gingival index: GI) and loss of periodontal attachment (LA). RESULTS: Contraceptive users of less than 2 years and 2-4 years duration (n=32, n= 17 respectively) and non-users (n=39) had similar oral hygiene levels; yet the contraceptive users had a significantly higher level of gingival inflammation, compared to the non-users (p<0.001; 1-way ANOVA). Usage of hormonal contraceptives for 2-4 years (n= 17) caused a significantly higher LA (p<0.001) compared to that of controls (n=39). CONCLUSIONS: Usage of contraceptive preparations containing oestrogen and progesterone resulted in hormonal changes similar to those seen in pregnancy, associated with increased prevalence of gingivitis. There was significantly higher LA with prolonged usage of hormonal contraceptives, compared with controls. PMID- 11034124 TI - Cytokine, elastase and oxygen radical release by Fusobacterium nucleatum activated leukocytes: a possible pathogenic factor in periodontitis. AB - Periodontitis is characterised by tissue destruction caused by reactive oxygen species (ROS) and proteolytic enzymes, which are released by the interaction between bacteria and phagocytes. We estimated the ability of Fusobacterium species to induce release of tissue destructive and proinflammatory mediators from in vitro activated peripheral leukocytes. ROS was measured with the nitroblue tetrazolium (NBT) method, elastase with a specific chromogenic substrate and cytokines, including interleukin 1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), and interleukin 8 (IL-8) with a sandwich ELISA method. Various clinical isolates of unopsonized Fusobacterium species stimulated the neutrophils to an increased NBT- reduction. IL-1beta, TNFalpha, IL-8 and elastase were released in significantly higher levels from neutrophils stimulated by Fusobacterium species. In conclusion, unopsonized Fusobacterium species can induce increased production of oxygen radicals, cytokines and elastase from leukocytes activated in vitro. PMID- 11034125 TI - Detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in dental plaque samples from children 2 to 12 years of age. AB - The purpose of this study was to detect the presence of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in plaque samples from 104 children, collected from their toothbrushes using a polymerase chain reaction (PCR). The age range of all subjects was 2-12 years. 21, 73 and 10 children with healthy gingiva, gingivitis and periodontitis respectively were selected. Plaque samples were collected from all erupted teeth sites using a sterile toothbrush. The mean concentration of DNA recovered from brushing plaque samples was approximately 660 microg/ml, which was sufficient for performing a PCR-based survey. Both A. actinomycetemcomitans and P. gingivalis were detected in the primary and mixed dentition. The prevalence of A. actinomycetemcomitans in healthy subjects was 4.8%, and those with gingivitis and periodontitis was 6.8% and 20.0% respectively, while the prevalence of P. gingivalis was 4.8% in healthy subjects, and 9.6% and 20.0% in those with gingivitis and periodontitis, respectively. Our survey, using a toothbrush, indicated that A. actinomycetemcomitans and P. gingivalis are rarely present in oral cavities of healthy children. PMID- 11034126 TI - Antibody responses against Porphyromonas gingivalis infection in patients with early-onset periodontitis. AB - BACKGROUND, AIMS: The aim of this study was to evaluate antibody responses against Porphyromonas gingivalis (P. gingivalis) infection in early-onset periodontitis (EOP) patients to elucidate further the host-parasite interactions in the pathogenesis of EOP. METHOD: 16 P. gingivalis-infected EOP and 20 adult periodontitis (AP) patients, and 18 periodontally healthy subjects (HS) participated in this study. Serum immunoglobulin G (IgG) antibody levels and avidities against extracted P. gingivalis whole cells were measured. The components of P. gingivalis outer membrane antigens (OMA) reacting to patients' sera were analysed from the molecular weights by Western blotting. Serum antibody levels against P. gingivalis lipopolysaccharide (LPS) were also measured. The ability of the patients' sera to block interleukin-1beta (IL-1beta) production by human mononuclear cells in response to P. gingivalis LPS was examined. RESULTS: Antibody levels were positively correlated with antibody avidities in both EOP and AP patients (r=0.91, r=0.72, p<0.0005, respectively), while not significantly so in HS (r=0.09). There was variability in the antigen recognition of P. gingivalis OMA in EOP and AP patients. Smear and 53-kDa protein were more frequently recognized by sera of EOP and AP patients rather than that of HS (p<0.05). The smear was partly diminished by absorption with P. gingivalis LPS, indicating the smear antigen was partly composed of LPS. There was high correlation between antibody levels against P. gingivalis whole-cell extracts and LPS in EOP and AP patients (r=0.81, p=0.0002, r=0.87, p<0.0001, respectively), while not significant in HS (r=0.22). The sera of EOP and AP patients with high IgG titre to P. gingivalis LPS blocked IL-1beta production more effectively than that of the patients with low IgG titre to P. gingivalis LPS. CONCLUSIONS: These results indicate that EOP patients' antibody response against P. gingivalis infection does not differ significantly from that of AP patients. The person-to person heterogeneous antibody production against P. gingivalis LPS could contribute to our understanding of the relationship between the defensive ability of EOP patients and their chronic infection with this pathogen. PMID- 11034127 TI - Associations of periodontal disease with femoral bone mineral density and estrogen replacement therapy: cross-sectional evaluation of US adults from NHANES III. AB - The objectives of this study were to evaluate the possible association of periodontal disease with (1) femoral bone mineral density (BMD), and (2) estrogen replacement therapy in a large sample of US adults (N= 11,655). The mean clinical attachment loss (CAL) per person was the main outcome variable. Based on the total BMD of the proximal femur and using the WHO diagnostic criteria, subjects were classified as having osteoporosis, osteopenia, or normal BMD. After adjusting for confounders, females with high calculus scores and low BMD had significantly more CAL than females with normal BMD and similar calculus scores (p<0.0001). No association was observed among women with low and intermediate levels of calculus. The greater CAL present among women with low BMD was associated with gingival recession. Patterns of findings were similar but equivocal among men, of whom only 66 were osteoporotic. After adjustment for possible confounders, postmenopausal women who reported having used estrogen replacement therapy presented significantly less mean CAL than those who never used estrogen. These findings indicate that in the presence of high calculus scores, females with osteoporosis are at increased risk for attachment loss and that this risk may be attenuated by the use of estrogen replacement therapy. PMID- 11034128 TI - Periodontal disease status during pregnancy and 3 months post-partum, in a rural population of Sri-Lankan women. AB - OBJECTIVES: The aim of this investigation was to study the effects of pregnancy on the periodontium, in a rural population of Sri-Lankan women. METHODS: The study group consisted of 47 pregnant women and 47 non-pregnant women who served as matched controls. All subjects were examined for plaque (plaque index: PLI), gingival condition (gingival index: GI) and loss of periodontal attachment (LA) levels, 4 x during the study, at 3-monthly intervals. RESULTS: Despite similar scores for plaque levels in both pregnant and non-pregnant women, the GI of pregnant women was significantly increased, during the 1st and 2nd trimesters compared to the controls (p<0.01, 2-way ANOVA). During the 3rd trimester, GI was further increased (p<0.001), but dropped at 3 months post-partum. Values for LA did not show significant differences from that of controls, during any of the stages of pregnancy. CONCLUSIONS: The results of this study show that pregnancy had an effect only on the gingivae and not on periodontal attachment levels. The effects of oestrogen and progesterone could give rise to a more florid response to the irritant effects of plaque, resulting in severe gingivitis. PMID- 11034129 TI - Are learning approaches and thinking styles related? A study in two Chinese populations. AB - This article presents the results of an investigation of the construct validity of J. B. Biggs's (1987) theory of learning approaches and of R. J. Sternberg's (1988) theory of thinking styles in two Chinese populations. The study is also an examination of the nature of the relations between the two theories. University students from Hong Kong (n = 854) and from Nanjing, mainland China (n = 215), completed the Study Process Questionnaire (J. B. Biggs, 1992) and the Thinking Styles Inventory (R. J. Sternberg & R. K. Wagner, 1992). Results indicated that both inventories were reliable and valid for assessing the constructs underlying their respective theories among both Hong Kong and Nanjing university students. Results also showed that the learning approaches and thinking styles are related in the hypothesized ways: The surface approach was hypothesized to be positively and significantly correlated with styles associated with less complexity, and negatively and significantly correlated with the legislative, judicial, liberal, and hierarchical styles. The deep approach was hypothesized to be positively and significantly correlated with styles associated with more complexity, and negatively and significantly correlated with the executive, conservative, local, and monarchic styles. Implications of these relations are discussed. PMID- 11034130 TI - Psychosocial predictors of acculturative stress in Mexican immigrants. AB - Psychosocial predictors of acculturative stress were examined in a sample of adult Mexican immigrants in Los Angeles. Bivariate and multivariate analyses revealed that family dysfunction, geographical separation from family, nonpositive expectations for the future, and low income levels were significantly related to elevated levels of acculturative stress. The findings suggest that family closeness, hopefulness for the future, and financial resources may provide a buffer against acculturative stressors experienced by migrating individuals. The findings highlight the importance of using culturally relevant clinical methods when assessing and treating immigrants and acculturating individuals. PMID- 11034131 TI - Psychosocial concomitants of loneliness among students of Cape Verde and Portugal. AB - This research is an examination of the relationship between loneliness and a number of psychosocial variables (e.g., affective state, cultural issues, gender, age) among adolescents and young adults from Cape Verde and Portugal. Two studies are presented. The participants in the first study were 285 adolescents from Cape Verde and 202 from Portugal, and in the second study there were 134 college students from Cape Verde and 112 from Portugal. The following instruments were administered to all the participants in both studies: The Revised UCLA Loneliness Scale (D. Russell, L. Peplau, & C. Cutrona, 1980), the Neuroticism Scale (J. Barros, 1999), the Optimism Scale (J. Barros, 1998), the Social Anxiety subscale (A. Fenigstein, M. Scheier, & A. Buss, 1975), and the Satisfaction With Life Scale (E. Diener, R. Emmons, R. Larsen, & S. Griffin, 1985). No ethnic or gender differences were found for loneliness between the two groups. For both groups, the most prominent predictors of loneliness were neuroticism and dissatisfaction with life. However, as expected, the adolescent group recorded higher scores for loneliness than did the college students. PMID- 11034132 TI - Perceptions of women's power as a function of position within an organization. AB - In the present study the authors investigated how people perceive women as a function of position within an organization. Introductory psychology students (N = 456) were given 1 of 6 vignettes depicting people in various occupations, along with an adjective checklist adapted from the Bem Sex Role Inventory (S. L. Bem, 1974). The students indicated on a Likert-type scale the adjectives they believed described the character in the paragraph they received. They were also given a power style scale by T. R. Hinkin and C. A. Schriesheim (1989) and were asked to rate the vignette characters' ability to use each different style (e.g., reward, expertise, referent, legitimate, coercive). The students rated women in high positions as more masculine than men and women in low positions and as masculine as men in high positions. There was little difference in power styles as a function of either the sex or the position level of the vignette character. PMID- 11034133 TI - Relationship satisfaction and dating experience: factors affecting future time orientation in relationships with the opposite sex. AB - This study was conducted to examine the effects of relationship satisfaction and dating experience on future time orientation in relationships with the opposite sex. Data from a sample of 226 undergraduate students from Middle East Technical University indicated that individuals who were highly satisfied in their relationships with the opposite sex tended to seek temporary relationships more and tended to be less future focused in their relationships than those who were less satisfied. Results also indicated that present dating experience did not show any significant effect on future time orientation; however, previous dating experience did show an effect in the interaction with relationship satisfaction and different factors of future time orientation. Implications of these results are discussed. PMID- 11034134 TI - Perceived overqualification and dimensions of job satisfaction: a longitudinal analysis. AB - The present study is an investigation of the effects of perceived overqualification on dimensions of job satisfaction. The data for this study came from a two-wave panel study of members of a midwestern American Postal Workers Union local. Job satisfaction was operationalized with 4 subscales from the Job Descriptive Index (P. Smith, L. Kendall, & C. Hulin, 1969). The following three hypotheses were tested: (a) Perceived overqualification will be negatively related to facets of job satisfaction; (b) there is stability in the test-retest correlations of facets of job satisfaction; and (c) the effects of perceived overqualification on facets of job satisfaction will not change from Time 1 to Time 2 because of adaptation. The cross-sectional results supported the hypotheses and suggested that perceived overqualification has a negative effect on job satisfaction. However, the relationships varied by dimension of perceived overqualification and dimension of job satisfaction. Future researchers of overqualification and dimensions of job satisfaction should consider relative deprivation as a source of work-related deprivations. PMID- 11034135 TI - Rethinking communication apprehension: a Myers-Briggs perspective. AB - This study is an examination of relationships between Myers-Briggs personality type preferences, based on Jungian theory, and communication apprehension. Results showed that participants who preferred introversion or sensing reported significantly higher levels of communication apprehension in general and across the group, dyadic, meeting, and public contexts than did participants who preferred extraversion or intuition. In addition, participants who preferred feeling reported higher levels of communication anxiety in the public context than those who preferred thinking. Findings support the assumption that communication apprehension is biologically based, suggest that the Myers-Briggs type preference framework offers an alternative way of understanding communication apprehension, and point out the need for new approaches to understanding the phenomenon of communication apprehension. PMID- 11034136 TI - Comparison of counseling and clinical psychology graduates on the examination for professional practice in psychology. AB - Doctoral recipients in clinical psychology scored higher than doctoral recipients in counseling psychology on the national licensing exam, the Examination for Professional Practice in Psychology. The largest discrepancies between the 2 groups were on Subtest I (diagnosis), Subtest II (intervention), and Subtest III (research). The differences were less large for Subtest IV (professional/ethical/legal issues) and for Subtest V (social application). The authors recommend that both counseling and clinical psychology graduate students take more course work in the department of the other discipline. PMID- 11034137 TI - Comparative aspects of invertebrate neuropeptides. AB - 1. We searched for bioactive peptides, most of which were considered to be neuropeptides, in various animals of several phyla. These peptides were compared with each other and with peptides identified by many other investigators. Consequently, we found that structures of neuropeptides are generally conserved in each phylum. 2. We also found some exceptional interesting aspects. First, there are a number of peptide groups whose members are distributed among several phyla. Second, there are many structural similarities between molluscan and annelidan peptides as if molluscs and annelids were the animals in a phylum. Third, certain toxic peptides of invertebrates are closely related to vertebrate neuropeptides. 3. In addition to the above phylogenetic aspects, we found some other interesting aspects. A wide structural variety of members of a peptide group is generally found in invertebrate species. Invertebrate muscles seem to be generally regulated not only by some or several classical non-peptidic neuromediators but also by various peptidic neuromediators. Peptides containing a D-amino acid residue are not rare. PMID- 11034138 TI - Mytilus inhibitory peptide (MIP) induces a Na+-activated K+-current in snail neurons. AB - Two microelectrode voltage-clamp and single-channel recordings were performed on D-cluster neurons of snail right parietal ganglion in order to study the properties of MIP-activated potassium current. It was found that the octapeptide member of the MIP-family, ASHIPRFVa elicits an outward current, which possesses all the properties characteristic for the hexapeptide(s) inward membrane response. The main component of the peptide elicited response is highly [K+]o dependent, however the response was attenuated in Na-free extracellular saline. The peptide elicited response was mimicked by raising the [Na+]i by pressure injection of Na+ into the cell. Single channel recordings indicated that MIP induced outward K-current is Na-dependent. The probability to find a channel in open state increases with increasing intracellular Na+-concentration. Excised inside-out patches obtained from D-neurons contained I(K(Na)) channels could be activated by exposure of the cytoplasmic face of the patch membrane to 40 mM Na+, and 40 mM Li+, as well. The single channel current amplitude at -60 mV is 15 pA and the single channel conductance is 212 pS between -80 and 0 mV. It was concluded that MIP's activate a novel type of K+-current in the snail neurons. This current is the Na-activated K+-current. The single channel properties of the MIP activated channel is in concert with I(K(Na)) data obtained on different vertebrate and invertebrate preparations. PMID- 11034139 TI - The presence and specificity of crustacean cardioactive peptide (CCAP) immunoreactivity in gastropod neurons. AB - CCAP-like immunoreactivity was detected in central neurons with small and medium diameters in both Helix and Lymnaea CNS. The intensity of immunoreactivity showed seasonal changes with a maximum intensity during spring. The overwhelming majority of nerve cell bodies exhibiting CCAP immunoreactivity is located in the cerebral and parietal ganglia of both Helix and Lymnaea. The neurons of pleural and buccal ganglia were devoid of CCAP-immunoreactivity. Following preabsorbtion of CCAP antibody in 1:15000 dilution with 10(-3) M CCAP or CCAP-related peptide (Helix -CCAP), immunoreactivity could not be observed in neurons, demonstrating the specificity of the antibody to CCAP-related molecules in both Helix and Lymnaea. PMID- 11034140 TI - Comparative pharmacology of feeding in molluscs. AB - 1. This paper reviews the role of transmitters in identified neurons of gastropod molluscs in generating and modulating fictive feeding. 2. In Lymnaea and Helisoma the 3 phase rhythm is generated by sets of interneurons which use acetylcholine for the N1 (protraction) phase, glutamate for the N2 (rasp) phase interneurons. The N3 interneurons are likely to use several different transmitters, of which one is octopamine. 3. In all the species examined, serotonin (5-HT) is released from giant cerebral cells. Other amines, including dopamine and octopamine, are present in the buccal ganglia and all these amines activate or enhance feeding. 4. Nitric oxide (NO), mostly originating from sensory processes, can also activate fictive feeding, but (at least in Lymnaea) may also be released centrally from buccal (B2) and cerebral neurons (CGC). 5. The central pattern generator for feeding is also modulated by peptides including APGWamide, SCP(B) and FMRFamide. 6. There is increasing evidence that most of these transmitters/modulators act on feeding neurons through second messenger systems- allowing them to act as longer-lasting neuromodulators of the feeding network. 7. Many of the transmitters are used in similar ways by each of the gastropods examined so far, so that their function in the CNS seems to have been conserved through evolution. PMID- 11034141 TI - The octopamine-containing buccal neurons are a new group of feeding interneurons in the pond snail Lymnaea stagnalis. AB - In the pond snail, Lymnaea stagnalis, the paired buccal ganglia contain 3 octopamine-immunoreactive neurons, which have previously been shown to be part of the feeding network. All 3 OC cells are electrically coupled together and interact with all the known buccal feeding motoneurons, as well as with all the modulatory and central pattern generating interneurons in the buccal ganglia. N1 (protraction) phase neurons: Motoneurons firing in this phase of the feeding cycle receive either single excitatory (depolarising) synaptic inputs (B1, B6 neurons) or a biphasic response (hyperpolarisation followed by depolarisation) (B5, B7 motoneurons). Protraction phase feeding interneurons (SO, N1L, NIM) also receive this biphasic synaptic input after OC stimulation. All of protraction phase interneurons inhibit the OC neurons. N2 (retraction) phase neurons: These motoneurons (B2, B3, B9, B10) and N2 interneurons are hyperpolarised by OC stimulation. N2 interneurons have a variable (probably polysynaptic) effect on the activity of the OC neurons. N3 (swallowing) phase: OC neurons are strongly electrically coupled to both N3 phase (B4, B4cluster, B8) motoneurons and to the N3p interneurons. In case of the interneuronal connection (OC<->N3) the electrical synapse is supplemented by reciprocal chemical inhibition. However, the synaptic connections formed by the OC neurons or N3p interneurons to the other members of the feeding network are not identical. CGC: The cerebral, serotonergic CGC neurons excite the OC cells, but the OC neurons have no effect on the CGC activity. In addition to direct synaptic effects, the OC neurons also evoke long-lasting changes in the activity of feeding neurons. In a silent preparation, OC stimulation may start the feeding pattern, but when fictive feeding is already occurring, OC stimulation decreases the rate of the fictive feeding. Our results suggest that the octopaminergic OC neurons form a sub population of N3 phase feeding interneurons, different from the previously identified N3p and N3t interneurons. The long-lasting effects of OC neurons suggest that they straddle the boundary between central pattern generator and modulatory neurons. PMID- 11034142 TI - The possible roles of the monoaminergic system in the feeding of the snail Helix pomatia. AB - The possible role of serotonin and dopamine in the feeding of Helix pomatia was studied applying immunocytochemical, biochemical, and behavioral techniques as well as bioassay experiments. Immunocytochemistry showed that dopamine-containing (thyrosin-hydroxylase-immunoreactive) neuronal elements of the crop and the gizzard belong to the intrinsic part, whereas serotonin-containing (serotonin immunoreactive) neuronal elements belong to the extrinsic part of the gastrointestinal nervous system. Bioassay studies on the spontaneous contractions of the crop and the gizzard showed that dopamine affected only the longitudinal muscle contractions by increasing both the tonus and contractility, whereas serotonin was effective on both the longitudinal and circular muscle contractions. Serotonin increased the tonus and contractility of longitudinal muscles in the crop but decreased them in the gizzard. Serotonin decreased the tonus and contractility of the circular muscles in the crop but increased them in the gizzard. Serotonin effects on the circular muscle of the gizzard were concentration dependent between a range of 10(-5) M-3 x 10(-5) M. HPLC measurements of monoamines in starved and satiated animals showed that the concentration of both dopamine and serotonin significantly decreased in both the CNS and different parts of the gastrointestinal tract of satiated animals, suggesting a significant monoamine liberation during feeding. The injection of monoamines (10(-3) and 10(-2) M) into the body cavity of starved animals showed that only dopamine was able to induce feeding whereas serotonin increased the general activity of the animals suggesting that the initiation of feeding is rather dopamine than serotonin dependent. PMID- 11034143 TI - Locomotor rhythms in the pond snail Lymnaea stagnalis: site of origin and neurotransmitter requirements. AB - 1. We have found that, in preparations of isolated CNS of the pond snail Lymnaea stagnalis, both serotonin (5HT) and dopamine (DA), as well as their respective precursors, 5HTP and DOPA, are effective in producing fictive intense (muscular) locomotion. 2. Phase-coupled to each of the above pedal rhythms are numerous identifiable pedal neurons including the respiratory interneuron RPeD1, thus suggesting interaction between networks responsible for locomotion and air breathing. 3. The novel DA/DOPA-dependent motor rhythm resembles the 5HT/5HTP dependent one in terms of activity of identifiable pedal neurons, being however considerably slower than the latter. 4. The results of transection experiments suggest that each of the rhythms is generated by a paired CPG lying entirely within the pedal ganglia. PMID- 11034144 TI - Morphology and physiology of pleural-to-buccal neurons coordinating defensive retraction with feeding arrest in the pond snail Lymnaea stagnalis. AB - 1. We have studied morphology, physiology and chemistry of a bilateral pair of pleural-to-buccal projecting neurons (PlB cells) of the pond snail Lymnaea stagnalis. Intracellular dye fills revealed axon arborization within neuropiles of ipsilateral pedal and cerebral ganglia, as well as in both buccal ganglia. Terminal axons of the left and right PlBs showed close proximity within the buccal commissure. 2. The left and right PlB neurons have been found electrotonically coupled and, sometimes, generating synchronous spikes. 3. The results show that two PlB cells operate as a single unit, and that paired buccal networks responsible for feeding rhythm are treated by the PlBs as a single target. PMID- 11034145 TI - Seasonal plasticity of synaptic connections between identified neurones in Lymnaea. AB - Here we investigate the synaptic connectivity of the giant dopamine containing neurone (RPeDI) of Lymnaea stagnalis during the winter months, in wild and laboratory bred animals. RPeD1 is one of the three neurones forming the respiratory central pattern generator (CPG) in Lymnaea and initiates ventilation under normal circumstances. Many of the follower cells of RPeD1 are ventilatory motor neurones. The connections of RPeD1 to its follower cells were investigated using standard intracellular recording techniques and dopamine was applied to the follower cells using a puffer pipette. During February and early March, RPeD1 was functionally disconnected from its follower cells, but connections reappeared towards the end of March. Most functionally disconnected cells failed to respond to applied dopamine, consistent with the hypothesis that there is down regulation of dopamine receptors in the follower cells of RPeD1 in the winter months. Behaviourally, Lymnaea that survive the winter, are not active at this time and do not indulge in lung ventilation, but stay quiescent. Thus functional disconnection of neurones from the CPG may be either a cause or a consequence of this change in behaviour. PMID- 11034146 TI - Modulation in firing pattern and oscillation in nerve cells of Lymnaea during network reconstruction. AB - The modulation and reconstruction of the cardio-respiratory neural circuit of Lymnaea stagnalis L. was compared to that of Helix ponatia L. where the input variation and signal molecules were found to have primary importance in network reorganization. From the cardio-respiratory circuit only neurons connected by afferent or efferent pathways to the peripheral chemosensory organ, the osphradium, were used. It was shown that, the general principles of the network reorganization is similar in the two species. The firing pattern of the neurons altered in Lymnaea depending on the input activation or presence of signal molecules in the vicinity of the neurons. The responses of the neurons to the same sensory information, originating from osphradium varied depending on their firing patterns. On central neurones the generation of phasic pattern and/or oscillation was an indicator of network disintegration leading to insensibility to the osphradial sensory inputs. Co-application of signal molecules (5HT, DA, GABA with opioid peptides) to the neurons caused a phasic firing pattern and/or oscillation leading to disintegration of one network and activation of another one. The effect of mu-opioid peptides on GABA-induced and voltage activated ion currents were shown to be the cellular target in reconstruction of neural networks in Lymnaea. The neural network reconstruction in vertebrate brain evoked by signal molecules can be compared to that observed in the identified network of Lymnaea stagnalis making this latter a useful model in further studies, too. PMID- 11034147 TI - Analysis and modulation of spike trains and oscillations in identified neural network of Lymnaea stagnalis L. AB - Identified neurons and members of functionally characterized clusters of the central nervous system of Lymnaea stagnalis L. were studied. Long-term spike trains (10-100 min) were collected using current clamp method. Firing patterns were analyzed by several mathematical tools e.g.: spike density function (SDF), interspike interval (ISI), Fourier-transform. Both the spike trains and oscillation of firing were modulated by 5HT (2 x 10(-5) M) and mu-opioid peptides (10(-5) M). Co-application of 5HT (2 x 10(-5) M) and DAGO (10(-5) M) turned the firing of the neurons (RPeD1 and A cells) opposite to the running pattern and eliminated the 0.3 Hz oscillation causing a new slow periodicity (0.1-0.05 Hz). PMID- 11034148 TI - Nonlinear activity of identified Lymnaea neurons. AB - Two long-lasting discharges of action potentials were recorded from a buccal cell of the pond snail, respectively, before and after superfusing the preparation with low-calcium solution. The corresponding sequences of interspike intervals were then analysed by the nonlinear prediction methods. The results yield evidence of a small but clear nonlinearity only in the second of analysed tachograms. This finding is evaluated and discussed. PMID- 11034149 TI - Membrane transduction pathway in the neuronal control of protein secretion by the albumen gland in Helisoma (Mollusca). AB - The albumen gland in Helisoma secretes a perivitelline fluid which surrounds each egg and is made up of several 66 kDa protein subunits and polysaccharide complexes. Forskolin, an adenylate cyclase activator, stimulated the secretion and release of the perivitelline fluid. An acidic extract of the central nervous system increased the intracellular concentration of cAMP in the albumen gland and this results in the release of the 66 kDa molecule and other proteins. Digestion of the brain extract with proteases abolished this activity, suggesting that the factor is a peptide. Cyclic AMP analogues and [BMX also stimulated the protein secretion in dose-dependent manner. Forskolin when added with the brain factor had an additive response. SQ22536, a non-competitive inhibitor of adenylate cyclase, inhibited brain extract dependent adenylate cyclase activity whereas aluminum fluoride, a G protein activator, was found to stimulate adenylate cyclase. Dopamine also stimulates protein secretion by the albumen gland and through the application of various agonists and antagonists of dopamine, it was established that the neurotransmitter acts via D1-like receptors by stimulating adenylate cyclase. PMID- 11034150 TI - Changes in molluscan neurosecretory cells during reproductive cessation: cause or effect? AB - The pond snail Lymnaea stagnalis has a maximum life span of about 22 months. At the age of about 250 days animals start to decrease egg laying activity and at about 500 days most animals ceased egg laying activity. At the age of cessation of egg laying the neurosecretory caudodorsal cells (CDCs) which control egg laying in Lymnaea exhibit reduced branching patterns. At this stage the cells still exhibit their physiological properties. CDCs still contain biologically active peptides and in the isolated CNS they still exhibit an afterdischarge upon electrical stimulation. Probably in the intact animal cessation of egg laying occurs because the CDCs are not activated anymore by natural egg laying inducing stimuli. In very old animals CDCs exhibit signs of degeneration indicating that cell death occur. After an extended period of no egg laying of Lymnaea physiological changes occur in the CDCs. CDCs from animals after an extended period of no egg laying failed to exhibit an afterdischarge. In such CDCs chemical and electrical coupling among the CDCs are reduced. Morphologically reduced CDCs predominantly fail to exhibit an afterdischarge. However, there are minimally branched CDCs that still could give an afterdischarge. Probably morphological reduction is not the only factor that defines afterdischarge failure. At present we suggest the following sequence of changes. 1. Morphological reduction of CDC branching patterns. 2. Cessation of egg laying. 3. Physiological changes in the CDCs resulting in afterdischarge failure. 4. Further morphological and physiological deterioration of CDCs. PMID- 11034151 TI - Pre- and postsynaptic effects of eugenol and related compounds on Helix pomatia L. neurons. AB - We showed how eugenol blocks the synaptic transmission and gave a possible interpretation how it inhibits the excitation-contraction coupling that several authors described previously. Eugenol acts both in the pre- and postsynaptic side of the neurons. It blocks the Ca2+-currents, decreases the membrane potential of the neurons, increases the inward resistance and decreases the GABA, ACh and glutamate evoked excitatory responses in submillimolar concentration. PMID- 11034152 TI - Amphetamine elicited potential changes in vertebrate and invertebrate central neurons. AB - The effects of amphetamine on potential changes in both vertebrate and invertebrate central neurons and factors affecting the potential changes were tested. The animals studied included mice, newborn rat and African snail. Seizure was elicited after lethal doses of d-amphetamine (75 mg/kg, i.p.) administration in mice. Repetitive firing of the action potentials were elicited after d amphetamine (1-30 microM) administration in thin thalamic brain slices of newborn rat. Bursting firing of action potentials in the giant African central RP4 neuron were also elicited after d-amphetamine or l-amphetamine (0.27 mM) administration. The amphetamine elicited bursting firing of action potentials was not blocked even after high concentrations of d-tubocurarine, atropine, haloperidol, hexamethonium administration. Therefore, the amphetamine elicited potential changes may not be directly related to the activation of the receptors of the neuron. The bursting firing of action potentials elicited by amphetamine occurred 20-30 min after amphetamine administration extracellularly, even after high concentrations of d-amphetamine administration (0.27, 1 mM). However, the bursting firing of potentials occurred immediately if amphetamine was administrated intracellularly at lower concentration. Extracellular application of ruthenium red, the calcium antagonist, abolished the amphetamine elicited bursting firing of action potentials. If intracellular injection of EGTA, a calcium ion chelator, or injection with high concentrations of magnesium, the bursting firing of potentials were immediately abolished. These results suggested that the active site of amphetamine may be inside of the neuron and the calcium ion in the neuron played an important role on the bursting of potentials. In two electrode voltage clamped RP4 neuron, amphetamine, at 0.27 mM, decreased the total inward and steady outward currents of the RP4 neuron. d-Amphetamine also decreased the calcium, Ia and the steady-state outward currents of the RP4 neuron. Besides, amphetamine elicited a negative slope resistance (NSR) if membrane potential was in the range of -50 to -10 mV. The NSR was decreased in cobalt substituted calcium free and sodium free solution. The effects of secondary messengers on the amphetamine elicited potential changes were tested. The bursting firing of action potentials elicited by amphetamine in central snail neurons decreased following extracellular application of H8 (N-(2-methyl-amino) ethyl-3-isoquinoline sulphonamide dihydrochloride), a specific protein kinase A inhibitor and anisomycin, a protein synthesis inhibitor. However, the bursting firing of action potentials were not affected after extracellular application of H7 (1,(5-isoquinolinesulphonyl)-2-methylpiperasine dihydrochloride), a specific protein kinase C (PKC) inhibitor, or intracellular application of GDPbetaS, a G protein inhibitor. The oscillation of membrane potential of the bursting activity was blocked after intracellular injection of 3'-deoxyadenosine, an adenylyl cyclase inhibitor. These results suggested that the bursting firing of action potentials elicited by d-amphetamine in snail neuron may be associated with the cyclic AMP second messenger system; on the other hand, it may not be associated with the G protein and protein kinase C activity. It is concluded that amphetamine elicited potential changes in both vertebrate and invertebrate central neurons. The changes are closely related to the ionic currents and second messengers of the neurons. PMID- 11034153 TI - Invertebrates in neurotoxicology. AB - Due to the relative simplicity of their nervous system, invertebrate animals were widely used in the past decades for studying the processes of excitability at membrane level, as well as the mechanisms of neuronal events and interneuronal communication. Parallel with investigating basic questions of neurobiology, lower animals have also been the object of toxicological studies, because simple invertebrate preparations with well-known physiological, biochemical and pharmacological characteristics proved to be excellent models for testing the action of natural and synthetic compounds important to human pharmaceutical research as well as in searching suitable chemicals for pest control. In the last ten-fifteen years with the growing interest towards environmental protection, a new field was opened for the application of invertebrates, namely, testing and monitoring the presence and harmful effects of anthropogenic toxic substances. Invertebrates are used today both as passive and as active biomonitors to detect and evaluate the level of pollution in a given ecosystem, and to study the effects and mechanisms of action of pollutants. Invertebrate nervous systems are suitable objects in clarifying the mechanisms of action of toxic chemicals at various levels of the neural regulation. Toxic influences can be reflected in behavioural alterations, by the modification of the function of different organs as well as the neural regulation, presented by examples on mussels and snails. In case of neurotoxicity, the targets of action are the elements of the nervous system. Alterations can occur in the permeability (ion channels) of the neuronal membrane influencing excitability, potential generation and propagation of nerve impulse, in the transmitter system (synthesis, release, elimination and binding to the receptors), in the interneuronal and neuroeffector connections responsible for co-ordinated and adequate responses to the internal and external challenges. For the future, it can be predicted that neurotoxicological research with new compounds cannot be effective without using invertebrate preparations, since, due to animal protection and restrictions in animal experimentation, the permission of the use of vertebrates is much more limited in most of the developed countries, and this trend is certainly to be expanded. PMID- 11034154 TI - Bioaccumulation and toxicity of aluminium in the pond snail at neutral pH. AB - The low solubility of aluminium (Al) at neutral pH means that it largely exists as colloidal particulates in aquatic systems. However, the pond snail Lymnaea stagnalis accumulates significant amounts of Al following exposure to water containing added Al (up to 500 microg l(-1)) at pH 7. This is accompanied by depression of behavioural activity (locomotion, feeding) which subsequently recovers, suggesting tolerance to the metal. The presence of silica ameliorates behavioural toxicity of Al, but does not prevent uptake of the metal. In vitro studies using the isolated central nervous system demonstrate toxicity at the cellular level. Extracellular application of Al (100 microM) led to membrane depolarisation, bursts of action potentials and action potential broadening. The chemical form in which Al is applied influences the extent of bioaccumulation and toxicity. Detailed knowledge of its solution chemistry is therefore essential. PMID- 11034155 TI - Time-dependent actions of aluminates on membrane and action potentials of snail neurons. AB - Aluminum (Al) is one of the elements, which is frequently subjected to experiments, however, the neurological observations with it are rather conflicting. The cause of this controversiality is not known but relates to some human disorders such as Alzheimer's disease and others as well. We studied the time-dependent actions of AlCl3 base solutions on resting membrane potential (Em), input resistance (Rin) and spike shape in giant neurons of the snail Helix pomatia L. at pH 7.7 and room temperature (22-24 degrees C) by use of intracellular technique. We reported significant differences in the effectiveness of the various Al solutions depending on the time of storage before use in the experiments (0, 2 and 6 days at room temperature). Freshly prepared and applied Al solutions caused a significant and dose-dependent depolarization with a concomitant decrease of Rin and the amplitude of the action potentials, but the 6 days solutions induced a hyperpolarization. Ouabain (0.1 mM) antagonized the hyperpolarization. The pH (7.1 or 7.7) and the time of the storage in combination also modified the direct membrane effects. Our experiments show that Al can induce differential membrane effects depending on the presence of various aluminum compounds. Namely, the predominate aluminum-monomer at pH 7.7 the Al(OH)4- might cause depolarization but the polynuclear aluminum complexes after polymerization of the monomers could hyperpolarize the neuronal membrane. We suppose, that the time-dependent equilibrium of various aluminum complexes plays a role in generating controversies in this field and emphasize again the importance of standardization of the experimental protocol. PMID- 11034156 TI - Heavy metals and neuroimmunomodulation in Mytilus edulis. AB - Immunocytes of mussels are the chief immune defense in these organisms. When an immunocyte becomes activated there is a conspicuous change in its morphology (i.e., from round to amoeboid) that can be quantified using image analytical tools. Active immunocytes will typically show larger perimeters and areas and a smaller shape factor. Immunocytes exposed to heavy metals become inactive (Cd, Hg and Pb) thus with smaller perimeters (e.g., Pb2+ 2 ppm: P = 69.72 micron) and areas (e.g., Pb2+ 2 ppm: A = 270 micron2) and larger shape factors (Pb2 2 ppm: SF = 0.65) than the unexposed control cells (alpha = 0.05). Xenobiotics may also interfere with neuroimmunomodulation processes such as nitric oxide (NO) release. The release of NO is catalyzed by a calcium dependent constitutive nitric oxide synthase (cNOS). Presently, we are exploring the effects of heavy metals and other pollutants on cNOS activity, measured as real time NO release, in immunocytes and pedal ganglia from M. edulis. Preliminary results suggest that immunocytes exposed to Pb2+ (5 ppm) cause NO release and does not seem to inhibit further NO release in the presence of morphine. The possible implications of NO mediated Pb2+ neurotoxicity are also explored. PMID- 11034157 TI - Evidence of opiates and opioid neuropeptides and their immune effects in parasitic invertebrates representing three different phyla: Schistosoma mansoni, Theromyzon tessulatum, Trichinella spiralis. AB - Studies done in our laboratories have demonstrated that the parasitic trematode, Schistosoma mansoni is capable of producing several proopiomelanocortin (POMC) peptides including beta-endorphin, adrenocorticotropin (ACTH), melanocyte stimulating hormone (alphaMSH) and enkephalin as well as morphine. Some of these opioids have been demonstrated to be immunosuppressive and may play an important part in immune evasion by these parasites. The parasitic nematode Trichinella spiralis also produces immune suppressive substances in vitro as well as causes immune suppression in its encysted stage in vivo. We recently have demonstrated the presence of morphine in both infected mice and in the nematode by HPLC and RIA. In a recent study of the leech Theromyzon tessulatum, we demonstrated the presence of proopiomelanocortin (POMC) and its derived peptides, ACTH and alphaMSH, in the immune tissues. The peptide was cloned and extensively purified by HPGPC and reversed-phase HPLC, and then sequenced. The 25.4 kDa protein was purified by gel permeation chromatography, anti-ACTH-affinity column separation followed by reversed-phase HPLC. Its amino acid determination was performed by Edman degradation, enzymatic treatments and electrospray mass spectrometry. The structure of the leech POMC-like precursor and its derived peptides demonstrates considerable amino acid sequence similarity with mammalian POMC. Taken together, these studies demonstrate that opiates and opioid neuropetides are present in invertebrates and their immunoregulatory actions have been conserved during evolution. The role of opiates and opioid peptides in immune and behavior modification of hosts is also discussed. PMID- 11034158 TI - The functional role of octopaminergic neurons in insect motor behavior. AB - The role of efferent, octopaminergic dorsal unpaired median (DUM) neurons in insects is examined by recording from them during motor behaviour. This population of neuromodulatory neurons is divided into sub-populations which are specifically activated or inhibited during ongoing motor behavior. These neurons are always activated in parallel to the respective motor circuits, and in addition to their modulatory effects on synaptic transmission may also cause metabolic changes in their target tissues. PMID- 11034159 TI - Tyramine injections reduce locust viability. AB - In the locust nervous system, tyramine is the direct precursor for octopamine synthesis and, as an octopamine analogue, it can activate octopamine receptors. Furthermore, the identification of specific tyramine receptors in Locusta migratoria and Drosophila melanogaster suggests that it is an important transmitter or modulator candidate. In this paper, we report that repeated tyramine injections reduced the viability of last instar larvae of Locusta and Schistocerca. In addition, a retardation of the last ecdysis was observed as a sublethal effect of the repeated tyramine treatment. Moreover, egg deposition by adult females was also retarded and/or drastically reduced. These effects show similarity to sublethal effects described for certain "insecticidal" octopamine receptor agonists, such as formamidines and phenyliminoimidazolidines. Since certain formamidine compounds were also shown to be agonists for the cloned tyramine receptors, it cannot be excluded that some lethal or sublethal consequences of tyramine administration are the result of an interaction with specific tyramine receptors. PMID- 11034160 TI - Acetylcholine, GABA, glutamate and NO as putative transmitters indicated by immunocytochemistry in the olfactory mushroom body system of the insect brain. AB - The distribution of glutamate, GABA and ChAT and of NADPH-diaphorase was immunocytochemically and histochemically investigated in the mushroom bodies of the cricket (Gryllus bimaculatus) and of the fruitfly (Drosophila melanogaster). Glutamate and NO are considered as putative transmitters of mushroom body Kenyon cell types. In the input area (calyces) of the mushroom bodies of Drosophila, the majority of olfactory projection neurons is stained with antibodies against ChAT. In addition, small GABA-immunoreactive presynaptic fibres of extrinsic neurons occur intermingled with the ChAT-immunoreactive elements in the calyces, and occupy distinct compartments in the stalk and lobes. Complex synaptic connectivity of putatively cholinergic and GABAergic extrinsic neurons and of Keyon cell dendrites within the calycal glomeruli of mushroom bodies is discussed. PMID- 11034161 TI - Social aggressiveness of female and subordinate male crickets is released by opiate receptor antagonist. AB - 1. In the cricket Gryllus bimaculatus, effects of opiate receptor antagonist naloxone, 9 or 30 microg per animal, on aggressive behavior were investigated. 2. Naloxone had no significant impact on aggression of isolated and dominant males. In contrast, the drug caused a dramatic release of social aggression in female and subordinate male crickets. 3. The results suggest that activity of the opioid system contributes to suppress aggression in subordinate males, as well as in females, during social contacts. PMID- 11034162 TI - Potencies of naturally-occurring AKH/RPCH peptides in Locusta migratoria in the acetate uptake assay in vitro and comparison with their potencies in the lipid mobilisation assay in vivo. AB - The biological potencies of a number of naturally-occurring octa- and decapeptides of the large AKH/RPCH family of peptides were determined in Locusta migratoria using the lipid-mobilising assay in vivo and the acetate uptake assay in vitro. The most potent of the newly-tested peptides in the in vitro assay, Phl CC, differs from the endogenous major locust peptide, Lom-AKH-I, only by an exchange of serine versus threonine at position 10. However, the most active peptide in the in vitro assay remains Lom-AKH-III. At the other extreme is the peptide Mem-CC which contains a tyrosine residue at position 4 rather than the more typical phenylalanine. This peptide is over 20,000 times less potent than Lom-AKH-III in the in vitro assay, and also results in an unusual dose-response curve in the in vivo assay. Only a few peptides are approximately equipotent in both assays, but mostly the bioanalogues have a higher potency in vitro. The majority of them are 2- to 10-fold more potent in vitro, but Ani-AKH and Lom-AKH III are 19- and 48-fold more potent. The results are discussed in relation to either the actions of proteases or of possible preferential binding of different receptors involved in the different assays. PMID- 11034164 TI - Distribution of Eisenia tetradecapeptide immunoreactive neurons in the nervous system of earthworms. AB - A detailed mapping of Eisenia-tetradecapeptide-immunoreactive neurons in the central and peripheral nervous system combined with quantitative morphological measurements was performed in Eisenia fetida and Lumbricus terrestris. In Eisenia, most labelled neurons were observed in the ganglia of the ventral cord (20.38% of the total cell number of the ganglion) and 15.67% immunoreactive cells occurred in the brain, while 6% of the neurons could be shown in the subesophageal ganglion. In the case of Lumbricus, most immunoreactive cells were found in the subesophageal ganglion (16.17%) and in the ventral ganglia (12.54%). The brain contained 122 ETP-immunoreactive cells (5.6%). The size of the immunoreactive cells varied between 35-75 microm. A small number of Eisenia tetradecapeptide immunoreactive fibres were seen to leave the ventral ganglia via segmental nerves, and labelled processes could also be observed in the stomatogastric system and the body wall. Labelled axon branches originating from the segmental nerves formed an immunoreactive plexus both between the circular and longitudinal muscle layer and on the inner surface of the longitudinal muscle layer. This inner plexus was especially rich in the setal sac. Among the superficial epithelial cells the body wall contained a significant number of immunoreactive cells. Only a few Eisenia-tetradecapeptide immunoreactive neurons and fibres occurred in the stomatogastric ganglia. In the enteric plexus the number of immunoreactive neurons and fibres decreased along the cranio-caudal axis of the alimentary tract. Eisenia-tetradecapeptide immunoreactive cells were also present among the epithelial cells in the alimentary canal. Some of these cells resembled sensory neurons in the foregut, while others showed typical secretory cell morphology in the midgut and hindgut. PMID- 11034163 TI - Physiological and pharmacological studies on nematodes. AB - Classical transmitters and neuroactive peptides act as transmitters or modulators within the central and peripheral nervous systems of nematodes, for example Ascaris suum and Caenorhabditis elegans. Acetylcholine (ACh) and gamma aminobutyric acid (GABA) are respectively the excitatory and inhibitory transmitters onto somatic body wall muscle while 5-hydroxytrypamine (5-HT) is the excitatory transmitter onto pharyngeal muscle. 5-HT also reduces ACh-induced contractions of somatic muscle and this action of 5-HT is mediated through activation of adenylate cyclase while that on pharyngeal muscle is mediated through inositol phosphate activation. Glutamate, dopamine and octopamine also have transmitter roles in nematodes. Neuroactive peptides of the RFamide family can excite somatic muscle, for example, AF-1 (KNEFIRFamide), AF-2 (KHEYLRFamide), AF-3 (AVPGVLRFamide) and AF-4 (GDVPGVLRFamide) or inhibit and relax this muscle, for example, PF-1 (SDPNFLRFamide), PF-2 (SADPNFLRFamide) and PF-4 (KPNlRFamide). In addition PF-3 (AF-8) (KSAYMRFamide) has a biphasic action on pharyngeal muscle, excitation followed by inhibition while AF-1 only inhibits this muscle. The peptide effects can be either pre- or postsynaptic or both and are likely to be mediated through second messenger systems. In addition these peptides modulate the action of classical transmitters, particularly ACh. PMID- 11034165 TI - Reorganization of peptidergic systems during brain regeneration in Eisenia fetida (Oligochaeta, Annelida). AB - After the extirpation of the brain reorganization of the peptidergic (FMRFamide, neuropeptide Y, proctolin) systems was studied in the newly forming cerebral ganglion of the annelid Eisenia fetida. During regeneration, all immunoreactive fibres appear on the 1st-2nd postoperative day. At the beginning of regeneration, immunoreactive neurons and fibres form a mixed structure in the wound tissue. On the 3rd postoperative day, FMRFamide positive and neuropeptide Y-immunoreactive, while on the 7th postoperative day proctolin-immunoreactive neurons appear in the loose wound tissue. From the 25th postoperative day a capsule gradually develops around it. The neurons of the preganglion move to the surface of the newly appearing preganglion. The number of these cells gradually increase, and by the 72th-80th postoperative days the localization and number of peptide immunoreactive neurons is similar to that in the intact one. The neurons of all examined peptidergic systems may originate from the neuroblasts, situated on the inner and outer surface of the intact ganglia (e.g. suboesophageal and ventral cord ganglia). In addition FMRFamide and proctolin immunoreactive neurons may take their derive by mitotic proliferation from the pharyngeal neurons, too. PMID- 11034166 TI - The central nervous system, its cellular organisation and development, in the tadpole larva of the ascidian Ciona intestinalis. AB - From its numerical composition, the central nervous system (CNS) of the ascidian larva is one of the simplest known nervous systems having a chordate plan. Fewer than 350 cells together constitute a caudal nerve cord, an interposed visceral ganglion containing motor circuits for swimming and, rostrally, an expanded sensory vesicle containing major sensory and interneuron regions of the CNS. Some cells are ependymal, with ciliated surfaces lining the neural canal, while others are clearly either sensory receptors or motoneurons, but most are distinguishable only on cytological grounds. Although reassignments between categories are still being made, there is evidence for determinancy of total cell number. We have made three-dimensional cell maps either from serial semithin sections, or from confocal image stacks of whole-mounted embryos and larvae stained with nuclear markers. Comparisons between the maps of neural tubes in embryos of successive ages, that is, between cells in one map and their progeny in older maps, enable us to follow the line of mitotic descent through successive maps, at least for the caudal neural tube. Details are clear for the lateral cell rows in the neural tube, at least until the latter contains approximately 320 cells, and somewhat for the dorsal cell row, but the ventral row is more complex. In the hatched larva, serial-EM reconstructions of the visceral ganglion reveal two ventrolateral fibre bundles at the caudalmost end, each of 10-12 axons. These tracts include at least five pairs of presumed motor axons running into the caudal nerve cord. Two pairs of axons decussate. Complementing this vertebrate feature in the CNS of the larval form of Ciona, we confirm that synapses form upon the somata and dendrites of its neurons, and that its motor tracts are ventral. PMID- 11034167 TI - Sensory organs of adult Amphilina foliacea (Amphilinida). AB - The distribution and morphological diversity of the sensory structures on the body surface, proboscis and caudal cavity of adult Amphilina foliacea have been investigated. Fifteen different types of receptors are described. Along with nonciliated and uniciated receptors bi- and multiciliated receptors have been found for the first time. The zonal distribution of the sensory structures and their coincidence within the same areas of the body surface have been revealed. The concentration of sensory structures at the posterior end may indirectly confirm a hypothesis of the unavailability of developed attachment disk in ancestors of amphilinids. PMID- 11034169 TI - Strategy for framework 6 PMID- 11034168 TI - The ultrastructure of glia-like cells in lateral nerve cords of adult Amphilina foliacea (Amphilinida). AB - The ultrastructure of main lateral nerve cords (MCs) of adult A. foliacea was studied. By examination of the serial sections it has been found that some glia like cells are located on a periphery of MC. The processes of glia-like cells surround MCs and penetrate into the cord and surround the group of adjacent axons and pairs of neurones. There is a fine extracellular matrix between processes of glia-like cells. The numerous tight junctions occur between processes. The difference between the perykaryon's cytoplasm of glia-like cells in anterior, posterior and central part of MCs was found. PMID- 11034170 TI - Pigs, society and opacity. PMID- 11034171 TI - US considers moves to relieve morale crisis at energy labs PMID- 11034172 TI - Novartis axes UK transplant centre. PMID- 11034174 TI - Reactor refit ignites debate on Japan's fusion strategy PMID- 11034173 TI - Researchers fight for access to Native American skeleton. PMID- 11034175 TI - US lab animals may win in lawsuit. PMID- 11034176 TI - Genomics initiative to decipher 10,000 protein structures. PMID- 11034177 TI - Age wins as Tokyo revamps retirement rules. PMID- 11034178 TI - Frozen body offers chance to travel back in time. PMID- 11034179 TI - Experts question precautionary approach. PMID- 11034180 TI - French lab seeks recipe for success PMID- 11034181 TI - Neuroscience. The sandman's secrets. PMID- 11034182 TI - Sloan Digital Sky Survey. A hundred million points of light PMID- 11034183 TI - Legal confusion over 'cloning' risks throwing baby out with bathwater. PMID- 11034185 TI - Loss of taxonomists is a threat to pest control. PMID- 11034184 TI - 'Benign neglect' of inner city led to TB epidemic. PMID- 11034186 TI - Research, innovation and politics. PMID- 11034187 TI - Magic beans. PMID- 11034188 TI - Programmable matter PMID- 11034189 TI - Uncertainty in climate change PMID- 11034191 TI - Meteorites. The long trip to Earth PMID- 11034190 TI - The logic of human learning. PMID- 11034192 TI - Immunology. Memory needs no reminders. PMID- 11034193 TI - Resolving a methane mystery. PMID- 11034194 TI - Optical clocks coming of age PMID- 11034196 TI - Ti3SiC2 has negligible thermopower PMID- 11034195 TI - Colour-enhancing protein in blue petals. PMID- 11034197 TI - Asymmetric toes aid underwater swimming. PMID- 11034198 TI - Wheat mutation rate after Chernobyl. PMID- 11034199 TI - Energy constraints on carnivore diet. PMID- 11034200 TI - Phytochromes and light signal perception by plants--an emerging synthesis. AB - For plants, the sensing of light in the environment is as important as vision is for animals. Fluctuations in light can be crucial to competition and survival. One way plants sense light is through the phytochromes, a small family of diverse photochromic protein photoreceptors whose origins have been traced to the photosynthetic prokaryotes. During their evolution, the phytochromes have acquired sophisticated mechanisms to monitor light. Recent advances in understanding the molecular mechanisms of phytochromes and their significance to evolutionary biology make possible an interim synthesis of this rapidly advancing branch of photobiology. PMID- 11034202 TI - Structural determinants of water permeation through aquaporin-1. AB - Human red cell AQP1 is the first functionally defined member of the aquaporin family of membrane water channels. Here we describe an atomic model of AQP1 at 3.8A resolution from electron crystallographic data. Multiple highly conserved amino-acid residues stabilize the novel fold of AQP1. The aqueous pathway is lined with conserved hydrophobic residues that permit rapid water transport, whereas the water selectivity is due to a constriction of the pore diameter to about 3 A over a span of one residue. The atomic model provides a possible molecular explanation to a longstanding puzzle in physiology-how membranes can be freely permeable to water but impermeable to protons. PMID- 11034201 TI - Id2 is a retinoblastoma protein target and mediates signalling by Myc oncoproteins. AB - In mammalian cells, Id proteins coordinate proliferation and differentiation. Id2 is a dominant-negative antagonist of basic helix-loop-helix transcription factors and proteins of the retinoblastoma (Rb) family. Here we show that Id2-Rb double knockout embryos survive to term with minimal or no defects in neurogenesis and haematopoiesis, but they die at birth from severe reduction of muscle tissue. In neuroblastoma, an embryonal tumour derived from the neural crest, Id2 is overexpressed in cells carrying extra copies of the N-myc gene. In these cells, Id2 is in molar excess of the active form of Rb. The overexpression of Id2 results from transcriptional activation by oncoproteins of the Myc family. Cell cycle progression induced by Myc oncoproteins requires inactivation of Rb by Id2. Thus, a dual connection links Id2 and Rb: during normal cell-cycle, Rb prohibits the action of Id2 on its natural targets, but oncogenic activation of the Myc-Id2 transcriptional pathway overrides the tumour-suppressor function of Rb. PMID- 11034203 TI - Efficient delivery of meteorites to the Earth from a wide range of asteroid parent bodies AB - Almost all meteorites come from asteroids, but identifying their specific parent bodies, and modelling their transport to the Earth, has proved to be difficult. The usual model of delivery through orbital resonances with the major planets has recently been shown to deplete the supply of meteorites much too rapidly to explain either the observed flux at the Earth, or the length of time the meteorites have spent in space (as measured by cosmic-ray exposure ages). Independently, it has been found that a force arising from anisotropically emitted thermal radiation from asteroidal fragments (the 'Yarkovsky effect') influences the fragments' orbits in important ways. Here we report the results of a detailed model for the transport of meteorites to the Earth, which includes the Yarkovsky effect and collisional evolution of the asteroidal fragments. We find that the Yarkovsky effect significantly increases the efficiency of the delivery of meteorites to the Earth, while at the same time allowing a much wider range of asteroids to contribute to the flux of meteorites. Our model also reproduces the observed distribution of cosmic-ray exposure ages of stony meteorites. PMID- 11034204 TI - Trapping and emission of photons by a single defect in a photonic bandgap structure AB - By introducing artificial defects and/or light-emitters into photonic bandgap structures, it should be possible to manipulate photons. For example, it has been predicted that strong localization (or trapping) of photons should occur in structures with single defects, and that the propagation of photons should be controllable using arrays of defects. But there has been little experimental progress in this regard, with the exception of a laser based on a single-defect photonic crystal. Here we demonstrate photon trapping by a single defect that has been created artificially inside a two-dimensional photonic bandgap structure. Photons propagating through a linear waveguide are trapped by the defect, which then emits them to free space. We envisage that this phenomenon may be used in ultra-small optical devices whose function is to selectively drop (or add) photons with various energies from (or to) optical communication traffic. More generally, our work should facilitate the development of all-optical circuits incorporating photonic bandgap waveguides and resonators. PMID- 11034205 TI - Colloidal ordering from phase separation in a liquid-crystalline continuous phase AB - Some binary mixtures exist as a single phase at high temperatures and as two phases at lower temperatures; rapid cooling therefore induces phase separation that proceeds through the initial formation of small particles and subsequent growth and coarsening. In solid and liquid media, this process leads to growing particles with a range of sizes, which eventually separate to form a macroscopically distinct phase. Such behaviour is of particular interest in systems composed of an isotropic fluid and a liquid crystal, where the random distribution of liquid-crystal droplets in an isotropic polymer matrix may give rise to interesting electro-optical properties. Here we report that a binary mixture consisting of an isotropic fluid and a liquid crystal forming the continuous phase does not fully separate into two phases, but self-organizes into highly ordered arrays of monodisperse colloidal droplet chains. We find that the size and spatial organization of the droplets are controlled by the orientational elasticity of the liquid-crystal phase and the defects caused by droplets exceeding a critical size. We expect that our approach to forming monodisperse, spatially ordered droplets in liquid crystals will allow the controlled design of ordered composites that may have useful rheological and optical properties. PMID- 11034206 TI - Patterning of polymer-supported metal films by microcutting AB - The ability to micropattern materials is of great importance for manufacturing advanced electronic, optical and mechanical devices ranging from displays to biosensors. For this purpose a variety of methods have been developed, including X-ray, electron-beam and photo-lithography, microcontact printing, embossing, micromouldings and cold welding. But these techniques are often of restricted applicability, involve a multitude of elaborate and cumbersome processing steps, or require aggressive chemistry. Here we describe a simple and versatile way to create well resolved metallic structures on polymer substrates, which is based on solid-state embossing of metal-coated polymer films. Ductility of both the metal layer and the polymer substrate permits the metal to be cut into surprisingly regular, micrometre-size structures. We illustrate the method by preparing patterned electrically conducting structures, highly efficient infrared polarizers and polarization-dependent colour filters. PMID- 11034207 TI - Quantifying the uncertainty in forecasts of anthropogenic climate change AB - Forecasts of climate change are inevitably uncertain. It is therefore essential to quantify the risk of significant departures from the predicted response to a given emission scenario. Previous analyses of this risk have been based either on expert opinion, perturbation analysis of simplified climate models or the comparison of predictions from general circulation models. Recent observed changes that appear to be attributable to human influence provide a powerful constraint on the uncertainties in multi-decadal forecasts. Here we assess the range of warming rates over the coming 50 years that are consistent with the observed near-surface temperature record as well as with the overall patterns of response predicted by several general circulation models. We expect global mean temperatures in the decade 2036-46 to be 1-2.5 K warmer than in pre-industrial times under a 'business as usual' emission scenario. This range is relatively robust to errors in the models' climate sensitivity, rate of oceanic heat uptake or global response to sulphate aerosols as long as these errors are persistent over time. Substantial changes in the current balance of greenhouse warming and sulphate aerosol cooling would, however, increase the uncertainty. Unlike 50-year warming rates, the final equilibrium warming after the atmospheric composition stabilizes remains very uncertain, despite the evidence provided by the emerging signal. PMID- 11034208 TI - Pressure-induced changes in the compression mechanism of aluminous perovskite in the Earth's mantle AB - Although aluminium is the fifth most abundant element in the Earth's mantle, its effect on the physical properties of perovskite, the main mineral phase in the lower mantle, has largely been ignored. It is becoming clear, however, that many properties of MgSiO3 perovskites are remarkably sensitive to small amounts of aluminium. In particular, perovskite with only 5 wt% Al2O3 has a bulk modulus 10% lower than that of the pure magnesian end-member. The increased compressibility may be due to the high concentrations of oxygen vacancies required to balance the charge of the aluminium; if so, this would have important consequences for the mantle, as aluminous perovskites could be weaker, have lower seismic velocities and be hosts for water. To test whether oxygen vacancies exist in aluminous perovskites, I have calculated the compressibility of end-member defect-bearing perovskites using ab initio methods. The results show that perovskites with oxygen vacancies do have significantly greater compressibilities than those without such vacancies. But the results also suggest that oxygen vacancies become unfavourable at high pressures, in which case only the physical properties of the shallow lower mantle would be affected by aluminium-with the deeper mantle retaining properties similar to those of aluminium-free perovskite. PMID- 11034209 TI - A marine microbial consortium apparently mediating anaerobic oxidation of methane. AB - A large fraction of globally produced methane is converted to CO2 by anaerobic oxidation in marine sediments. Strong geochemical evidence for net methane consumption in anoxic sediments is based on methane profiles, radiotracer experiments and stable carbon isotope data. But the elusive microorganisms mediating this reaction have not yet been isolated, and the pathway of anaerobic oxidation of methane is insufficiently understood. Recent data suggest that certain archaea reverse the process of methanogenesis by interaction with sulphate-reducing bacteria. Here we provide microscopic evidence for a structured consortium of archaea and sulphate-reducing bacteria, which we identified by fluorescence in situ hybridization using specific 16S rRNA-targeted oligonucleotide probes. In this example of a structured archaeal-bacterial symbiosis, the archaea grow in dense aggregates of about 100 cells and are surrounded by sulphate-reducing bacteria. These aggregates were abundant in gas hydrate-rich sediments with extremely high rates of methane-based sulphate reduction, and apparently mediate anaerobic oxidation of methane. PMID- 11034210 TI - Modern freshwater microbialite analogues for ancient dendritic reef structures. AB - Microbialites are organosedimentary structures that can be constructed by a variety of metabolically distinct taxa. Consequently, microbialite structures abound in the fossil record, although the exact nature of the biogeochemical processes that produced them is often unknown. One such class of ancient calcareous structures, Epiphyton and Girvanella, appear in great abundance during the Early Cambrian. Together with Archeocyathids, stromatolites and thrombolites, they formed major Cambrian reef belts. To a large extent, Middle to Late Cambrian reefs are similar to Precambrian reefs, with the exception that the latter, including terminal Proterozoic reefs, do not contain Epiphyton or Girvanella. Here we report the discovery in Pavilion Lake, British Columbia, Canada, of a distinctive assemblage of freshwater calcite microbialites, some of which display microstructures similar to the fabrics displayed by Epiphyton and Girvanella. The morphologies of the modern microbialites vary with depth, and dendritic microstructures of the deep water (> 30 m) mounds indicate that they may be modern analogues for the ancient calcareous structures. These microbialites thus provide an opportunity to study the biogeochemical interactions that produce fabrics similar to those of some enigmatic Early Cambrian reef structures. PMID- 11034211 TI - Minimization of Boolean complexity in human concept learning. AB - One of the unsolved problems in the field of human concept learning concerns the factors that determine the subjective difficulty of concepts: why are some concepts psychologically simple and easy to learn, while others seem difficult, complex or incoherent? This question was much studied in the 1960s but was never answered, and more recent characterizations of concepts as prototypes rather than logical rules leave it unsolved. Here I investigate this question in the domain of Boolean concepts (categories defined by logical rules). A series of experiments measured the subjective difficulty of a wide range of logical varieties of concepts (41 mathematically distinct types in six families--a far wider range than has been tested previously). The data reveal a surprisingly simple empirical 'law': the subjective difficulty of a concept is directly proportional to its Boolean complexity (the length of the shortest logically equivalent propositional formula)--that is, to its logical incompressibility. PMID- 11034212 TI - Skin abnormalities generated by temporally controlled RXRalpha mutations in mouse epidermis. AB - Nuclear receptors for retinoids (RARs) and vitamin D (VDR), and for some other ligands (TRs, PPARs and LXRs), maybe critical in the development and homeostasis of mammalian epidermis. It is believed that these receptors form heterodimers with retinoid X receptors (RXRs) to act as transcriptional regulators. However, most genetic approaches aimed at establishing their physiological functions in the skin have been inconclusive owing either to pleiotropic effects and redundancies between receptor isotypes in gene knockouts, or to equivocal interpretation of dominant-negative mutant studies in transgenic mice. Moreover, knockout of RXRalpha, the main skin RXR isotype, is lethal in utero before skin formation. Here we have resolved these problems by developing an efficient technique to create spatiotemporally controlled somatic mutations in the mouse. We used tamoxifen-inducible Cre-ER(T) recombinases to ablate RXRalpha selectively in adult mouse keratinocytes. We show that RXRalpha has key roles in hair cycling, probably through RXR/VDR heterodimers, and in epidermal keratinocyte proliferation and differentiation. PMID- 11034213 TI - Memory B-cell persistence is independent of persisting immunizing antigen. AB - Immunological memory in the antibody system is generated in T-cell-dependent responses and carried by long-lived memory B cells that recognize antigen by high affinity antibodies. But it remains controversial whether these B cells represent true 'memory' cells (that is, their maintenance is independent of the immunizing antigen), or whether they are a product of a chronic immune response driven by the immunizing antigen, which can be retained in the organism for extended time periods on the surface of specialized antigen-presenting cells (follicular dendritic cells). Cell transfer experiments provided evidence in favour of a role of the immunizing antigen; however, analysis of memory cells in intact animals, which showed that these cells are mostly resting and can persist in the absence of detectable T-cell help or follicular dendritic cells, argued against it. Here we show, by using a genetic switch mediated by Cre recombinase, that memory B cells switching their antibody specificity away from the immunizing antigen are indeed maintained in the animal over long periods of time, similar to cells retaining their original antigen-binding specificity. PMID- 11034214 TI - A common E2F-1 and p73 pathway mediates cell death induced by TCR activation. AB - Strong stimulation of the T-cell receptor (TCR) on cycling peripheral T cells causes their apoptosis by a process called TCR-activation-induced cell death (TCR AICD). TCR-AICD occurs from a late G1 phase cell-cycle check point independently of the 'tumour suppressor' protein p53. Disruption of the gene for the E2F-1 transcription factor, an inducer of apoptosis, causes significant increases in T cell number and splenomegaly. Here we show that T cells undergoing TCR-AICD induce the p53-related gene p73, another mediator of apoptosis, which is hypermethylated in lymphomas. Introducing a dominant-negative E2F-1 protein or a dominant-negative p73 protein into T cells protects them from TCR-mediated apoptosis, whereas dominant-negative E2F-2, E2F-4 or p53 does not. Furthermore, E2F-1-null or p73-null primary T cells do not undergo TCR-mediated apoptosis either. We conclude that TCR-AICD occurs from a late G1 cell-cycle checkpoint that is dependent on both E2F-1 and p73 activities. These observations indicate that, unlike p53, p73 serves to integrate receptor-mediated apoptotic stimuli. PMID- 11034215 TI - Role for the p53 homologue p73 in E2F-1-induced apoptosis. AB - The transcription factor E2F-1 induces both cell-cycle progression and, in certain settings, apoptosis. E2F-1 uses both p53-dependent and p53-independent pathways to kill cells. The p53-dependent pathway involves the induction by E2F-1 of the human tumour-suppressor protein p14ARF, which neutralizes HDM2 (human homologue of MDM2) and thereby stabilizes the p53 protein. Here we show that E2F 1 induces the transcription of the p53 homologue p73. Disruption of p73 function inhibited E2F-1-induced apoptosis in p53-defective tumour cells and in p53-/- mouse embryo fibroblasts. We conclude that activation of p73 provides a means for E2F-1 to induce death in the absence of p53. PMID- 11034216 TI - S-RNase uptake by compatible pollen tubes in gametophytic self-incompatibility. AB - Many flowering plants avoid inbreeding through a genetic mechanism termed self incompatibility. An extremely polymorphic S-locus controls the gametophytic self incompatibility system that causes pollen rejection (that is, active arrest of pollen tube growth inside the style) when an S-allele carried by haploid pollen matches one of the S-alleles present in the diploid style. The only known product of the S-locus is an S-RNase expressed in the mature style. The pollen component to this cell-cell recognition system is unknown and current models propose that it either acts as a gatekeeper allowing only its cognate S-RNase to enter the pollen tube, or as an inhibitor of non-cognate S-RNases. In the latter case, all S-RNases are presumed to enter pollen tubes; thus, the two models make diametrically opposed predictions concerning the entry of S-RNases into compatible pollen. Here we use immunocytochemical labelling of pollen tubes growing in styles to show accumulation of an S-RNase in the cytoplasm of all pollen-tube haplotypes, thus providing experimental support for the inhibitor model. PMID- 11034217 TI - The large-scale organization of metabolic networks. AB - In a cell or microorganism, the processes that generate mass, energy, information transfer and cell-fate specification are seamlessly integrated through a complex network of cellular constituents and reactions. However, despite the key role of these networks in sustaining cellular functions, their large-scale structure is essentially unknown. Here we present a systematic comparative mathematical analysis of the metabolic networks of 43 organisms representing all three domains of life. We show that, despite significant variation in their individual constituents and pathways, these metabolic networks have the same topological scaling properties and show striking similarities to the inherent organization of complex non-biological systems. This may indicate that metabolic organization is not only identical for all living organisms, but also complies with the design principles of robust and error-tolerant scale-free networks, and may represent a common blueprint for the large-scale organization of interactions among all cellular constituents. PMID- 11034218 TI - Western researchers reap rich rewards in eastern institutions. PMID- 11034219 TI - Effects of isokinetic strength training on concentric and eccentric torque development in the ankle dorsiflexors of older adults. AB - This study quantifies concentric and eccentric dorsiflexor muscle torque at various movement velocities and determines the neuromuscular effects from short term practice of repeated maximal voluntary contractions using an isokinetic resisted exercise program in older adults. Twenty-eight subjects (76.3+/-4.6 years) trained concentric and eccentric dorsiflexion 3 days per week for 2 weeks at 30 degrees, 90 degrees, and 180 degrees/s through 40 degrees of dorsiflexion range of motion. Peak torque, rate of torque development, surface electromyography, and strength curves at each velocity were compared before and after training. Increases were found for concentric (27%) and eccentric (20%) dorsiflexor peak torque and concentric rate of torque production (20%-53%) across all movement velocities (p < .01). Training also significantly increased dorsiflexor concentric (64%) and eccentric (55%) surface electromyography; taken together, this is evidence of apparent adaptation of neural factors in older adults with short-term training. These findings support that eccentric and concentric strength training enhances the control and production of ankle muscle dorsiflexor torque in older adults, with implications for improving functional mobility of the ankle joint. PMID- 11034220 TI - Motor slowing and parkinsonian signs in aging rhesus monkeys mirror human aging. AB - Motor slowing is a universal feature of human aging, and parkinsonian signs are commonly expressed in human senescence. In the present study, age-associated declines in motor functions in 31 female rhesus monkeys were quantified by activity monitors and an automated test panel, and the incidence of parkinsonian signs was scored using a movement dysfunction assessment scale. Activity levels in middle-aged monkeys (12-17 years old) were less than half that of young animals (5-8 years old) and were further depressed in aged monkeys (21-27 years old). Movement dysfunction scores increased significantly with increasing age. Two or more parkinsonian signs were exhibited by 20% of the middle-aged monkeys and 36% of the aged monkeys. Slowing performance times on fine-motor hand tasks correlated significantly with increasing age. Motor learning was seen in all age groups, but improved faster in the young monkeys. The data suggest that aging rhesus monkeys provide an appropriate model to analyze the biological processes leading to motor slowing and the expression of parkinsonian signs in human senescence. PMID- 11034221 TI - Quadriceps muscle function in relation to habitual physical activity and VO2max in men and women aged more than 65 years. AB - The relationship of quadriceps maximal muscle power (Pmax), corresponding optimal shortening velocity (v(opt)), and relative fatigability (Pmax%D) to maximal oxygen uptake (VO2max) and habitual physical activity (PA) was examined in healthy community-dwelling subjects (29 women and 25 men) aged more than 65 years old. PA was evaluated by a questionnaire and expressed using two activity indices: mean habitual daily energy expenditure (MHDEE) and the daily energy expenditure corresponding to leisure time sports activities (Sports Activity). In women, Pmax correlated positively with VO2max (r = .56) and with Sports Activity (rho = .41). Both Sports Activity and Pmax were significant independent predictors of VO2max and accounted for 62% of variance in VO2max. In men, v(opt) was significantly negatively related to MHDEE (r = -.59) and to Sports Activity (rho = -.40). Neither in women nor in men was Pmax%D correlated with VO2max or PA indices. The different relationship of Pmax and v(opt) with VO2max and PA indices suggests that habitual PA may be sufficient in active older women, but not in men, to positively influence quadriceps muscle function. These gender differences may suggest different approaches in exercise programming for elderly women and men. PMID- 11034222 TI - Stimulation of prostaglandin E2 and interleukin-1beta production from old rat periodontal ligament cells subjected to mechanical stress. AB - Although the severity of periodontal disease is known to be affected by the age of the host, the pathological role of aging in periodontal disease, especially that attributable to trauma from occlusion, has not been well characterized. Prostaglandin (PG)E2 and interleukin (IL)-1beta are key mediators involved in periodontal diseases, potent stimulators of bone resorption, and are produced by human periodontal ligament (PDL) cells in response to mechanical stress. To investigate age-related changes in the biosynthetic capacity of PGE2 and IL-1beta in PDL cells, we examined the effects of in vivo aging with mechanical tension on PGE2 and IL-1beta expression by rat PDL cells. PDL cells obtained from the incisors of 6-week (young) and 60-week (old) rats were cultured on flexible bottomed culture plates. The cells were deformed by causing a 9% or 18% increase in surface area at 6 cycles per minute for 1 to 5 days. We found an approximately twofold increase in PGE2 and IL-1beta production by old PDL cells subjected to mechanical tension compared with that by young cells, although the constitutive levels were similar in both. The expression of cyclooxygenase (COX)-2 and IL 1beta mRNA (messenger ribonucleic acid) was enhanced by mechanical tension as determined by use of reverse transcription-polymerase chain reaction (RT-PCR), whereas COX-1 and IL-1beta-converting enzyme mRNA remained unchanged. It is possible that the large amount of PGE2 and IL-1beta produced by PDL cells from an aged host in response to mechanical force may be positively related to the acceleration of alveolar bone resorption. PMID- 11034223 TI - Effect of long-term dietary antioxidant supplementation on influenza virus infection. AB - This study compared the effect of vitamin E on the course of influenza infection with that of other antioxidants. (In a previous study we showed that short-term vitamin E supplementation significantly decreased pulmonary viral titer in influenza-infected old mice). Eighteen-month-old C57BL/6NCrlBR mice were fed one of the following semisynthetic diets for 6 months: control, vitamin E supplemented, glutathione supplemented, vitamin E and glutathione supplemented, melatonin supplemented, or strawberry extract supplemented. After influenza virus challenge, mice fed vitamin E-supplemented diet had significantly lower pulmonary viral titers compared to those fed the control diet (10(2.6) vs 10(4.0), p < .05) and were able to maintain their body weight after infection (1.8+/-0.9 g weight loss/5 days postinfection in vitamin E group vs 6.8+/-1.4 g weight loss/5 days postinfection in control group, p < .05). Other antioxidants did not have a significant effect on viral titer or weight loss. There was a significant inverse correlation of weight loss with food intake (r = -.96, p < .01), indicating that the observed weight changes were mainly due to decreased food intake. Pulmonary interleukin (IL)-6, IL-1beta, and tumor necrosis factor (TNF)-alpha levels increased significantly postinfection. The vitamin E group had lower lung IL-6 and TNF-alpha levels following infection compared to the control group. In addition, there was a significant positive correlation between weight loss and lung IL-6 (r = .77, p < .01) and TNF-alpha (r = .68, p < .01) levels. Because IL 6 and TNF-alpha have been shown to contribute to the anorexic effect of infectious agents, the prevention of weight loss by vitamin E might be due to its reduced production of IL-6 and TNF-alpha following infection. Thus, among the antioxidants tested, only vitamin E was effective in reducing pulmonary viral titers and preventing an influenza-mediated decrease in food intake and weight loss. Other dietary antioxidant supplementations that reduced one or more measures of oxidative stress (4-hydroxynonenal, malondialdehyde, and hydrogen peroxide) did not have an effect on viral titer, which suggests that, in addition to its antioxidant activity, other mechanisms might be involved in vitamin E's beneficial effect on lowering viral titer and preventing weight loss. PMID- 11034224 TI - Use of muscle functional magnetic resonance imaging with older individuals. AB - Muscle functional magnetic resonance imaging (mfMRI) has been widely used to study muscle recruitment in exercise in young healthy subjects, but has not been validated or used with older subjects. This study validates and demonstrates the use of mfMRI in older subjects. Subjects consisted of apparently healthy sedentary younger (n = 7) and older (n = 6) women. Proton transverse relaxation (T2)-weighted MRI scans were obtained of the quadriceps femoris at rest and immediately following three bouts of knee extension exercise (50%, 75%, and 100% of untrained 5 x 10 repetition maximum [RM]). Older subjects performed knee extension training for 12 weeks and repeated the MRI scan protocol using the same absolute loads. Training induced a 13% increase in 1 RM and a 25% increase in 5 x 10 RM. Older subjects had higher resting T2 values compared with younger subjects; however, the T2 response to exercise (slope) was similar among groups (young = 0.063+/-0.003, older untrained = 0.055+/-0.011, older trained = 0.053+/ 0.008; p > .05). In all cases, T2 increased linearly with load. Trained older subjects showed a lower T2 response when lifting the same absolute load compared with before training, which is consistent with results previously obtained from young subjects. In the older population, mfMRI is appropriate for use and offers benefits over other technologies. PMID- 11034225 TI - The medication list--a portrait of a patient's health. PMID- 11034226 TI - One-year follow-up of medication management capacity in highly functioning older adults. AB - BACKGROUND: We tested the hypothesis that impairment in the ability to take medication independently predicts early functional decline. METHODS: A 12-month, prospective cohort study was performed at two continuing-care retirement facilities using the Drug Regimen Unassisted Grading Scale (DRUGS). This geriatric screening tool utilizes a stepwise progression of four tasks: (i) identification, (ii) access. (iii) dosage, and (iv) timing. RESULTS: Forty-seven (86%) of the eligible participants completed the 12-month follow-up assessment; three were transferred to skilled nursing facilities. The mean age at study entry was 84.2+/-5.1 years; 72% of the participants were women, and 68% were college educated. At 12 months there was a decline in the Mini-Mental State Examination (MMSE) score (p = .029), an increase in the timed "Up and Go" test (p = .023), and a decline in the DRUGS score (p .029). Nine (18%) of the participants resided in assisted- versus independent-living situations compared with three participants (5%) at study entry (p = .031). Both 12-month DRUGS score and 12 month self-reported medication management capacity were associated with 12-month MMSE (p = .0001 and p = .019, respectively). Baseline DRUGS score was associated with 12-month MMSE and Geriatric Depression Scale scores (p = .0002 and p = .002, respectively). Both baseline DRUGS score and self-reported medication management capacity were also associated with residence in assisted-living communities at 6 months (p = .029 and p = .040, respectively). MMSE was not associated with any of the clinical outcomes. PMID- 11034227 TI - Polypharmacy and hospitalization among older home care patients. AB - BACKGROUND: One of the major goals of home care is the prevention of hospitalization. The objective of this study was to examine the relation between medication use (number, type, and inappropriateness) and hospitalization among home care patients older than 65 years. METHODS: A retrospective chart review of 833 discharged older home care patients was performed. These patients were consecutive discharges from a single home care agency who either (a) returned to independent self-care or care of the family (S/F Care group) or (b) were admitted to the hospital (Hospitalized group). Medication assessment within these two groups included total number of medications (prescription and nonprescription); degree of polypharmacy (percentage of patients taking 5 or more, 7 or more, and 10 or more medications); and prevalence for different types of medications, including different types of inappropriate medications. Inappropriate medications were designated according to a list that was previously developed through a modified Delphi consensus technique by a panel of 13 experts in geriatric pharmacology and has been utilized in other studies. Student's t test was used for continuous variables and chi-square test was used for categorical variables to evaluate for differences between the S/F Care group and the Hospitalized group (p <.05). For comparisons of types of medications, p < .01 was used for significant differences, because of the high number of comparisons made. RESULTS: Of 833 discharges, 644 (77.3%) returned to self-care or care of the Family (S/F Care group) and 189 (22.7%) were hospitalized. The Hospitalized group, compared with the S/F Care group, was taking a higher number of medications (mean +/- SD: 6.6+/-3.9 vs 5.7+/-3.4, p = .004), and had a higher percentage of patients taking 7 or more medications (46% vs 26%, p = .002) and 10 or more medications (21% vs 10%, p = .005), but not 5 or more medications. Only three types of medications were more commonly used among patients in the Hospitalized group than among patients in the S/F Care group: clonidine (4.2% vs 1.1%, p = .004); mineral supplements (23.8% vs 14.8%, p = .003); and metoclopramide (5.8% vs 2.0%, p = .006). The Hospitalized group had a lower percentage of patients taking inappropriate medications than did the S/F Care group (20% vs 27%, p = .040), but none of the types of inappropriate medications was used more often in either group. CONCLUSIONS: This study shows a relationship between high levels of polypharmacy and hospitalization. Although it cannot be determined from this study whether a higher number of medications was an indicator of sicker patients at risk for hospitalization, or whether a higher number of medications might have directly led to hospitalization, polypharmacy should still be considered a marker for older home care patients for whom prevention of hospitalization is the goal. PMID- 11034228 TI - Geriatrics grand rounds: Eve's rib, or a revisionist view of osteoporosis in men. AB - It is widely accepted that estrogen withdrawal following menopause predisposes women to accelerated bone loss and increased risk of developing osteoporosis. Although osteoporosis is a significant public health problem for aging men as well as women, the cause of osteoporosis in men remains largely unknown. A substantial number of men with osteoporosis present with bone loss secondary to conditions associated with reduced gonadal steroid hormone levels. Although hypogonadism is related to bone loss in men, and androgen levels decline with age in men, it is not at all clear that reduced androgen levels are related to bone loss in older men. What, then, is the role of gonadal steroids in osteoporosis in men? This review focuses on recent research--including clinical investigations of men with genetic disorders of estrogen action, basic biomedical studies of estrogen receptor "knockout" mice, and population-based comparisons of bone density with gonadal steroids in older men--leading to the surprising conclusion that estrogen plays a vital role in maintenance of bone in men as well as in women. Possible mechanisms whereby reduced estrogen levels might result in bone loss in both sexes are also reviewed, as are potential therapeutic implications of a role for estrogen in osteoporosis in men. PMID- 11034229 TI - Improved functional outcomes following exercise rehabilitation in patients with intermittent claudication. AB - BACKGROUND: The purposes of this study were to identify predictors of increased claudication distances following exercise rehabilitation in peripheral arterial occlusive disease (PAOD) patients with intermittent claudication and determine whether improved claudication distances translated into increased free-living daily physical activity in the community setting. METHODS: Sixty-three patients were recruited (age, 68+/-1 years, mean +/- standard error). Patients were characterized on treadmill claudication distances, walking economy, peripheral circulation, cardiopulmonary function, self-perceived ambulatory function, body composition, baseline comorbidities, and free-living daily physical activity before and after a 6-month treadmill exercise program. RESULTS: Exercise rehabilitation increased distance to onset of claudication pain by 115% (178+/-22 m to 383+/-34 m; p < .001) and distance to maximal claudication pain by 65% (389+/-29 m to 641+/-34 m; p < .001). The increased distance to onset of pain was independently related to a 27% increase in calf blood flow (r = .42, p < .001) and to baseline age (r = -.26, p < .05), and the increased distance to maximal pain was predicted by a 10% increase in peak oxygen uptake (r = .41, p < .001) and by a 10% improvement in walking economy (r = -.34, p < .05). Free-living daily physical activity increased 31% (337+/-29 kcal/day to 443+/-37 kcal/day; p < .001) and was related to the increases in treadmill distances to onset (r = .24, p < .05) and to maximal pain (r = .45, p < .001). CONCLUSIONS: Increased claudication distances following exercise rehabilitation are mediated through improvements in peripheral circulation, walking economy, and cardiopulmonary function, with younger patients having the greatest absolute ambulatory gains. Furthermore, improved symptomatology translated into enhanced community-based ambulation. PMID- 11034230 TI - The use of a simulated environment (easy street) to retrain independent living skills in elderly persons: a randomized controlled trial. AB - BACKGROUND: Older adults who receive training for functional skills in contextually appropriate environments may show greater functional improvement than persons trained in a traditional environment. Functionally limited older adults receiving training in contextually appropriate environments (simulated home and community settings) may show greater improvement in activities of daily living (ADL) than persons trained in a traditional manner. METHODS: Eighty-eight patients from a day hospital, aged 65 years or older, were randomized to either receive rehabilitation in a simulated environment (Easy Street) or in a gymnasium setting. Rehabilitation focused on retraining functional skills in a contextually appropriate environment (Easy Street) or in a traditional setting (gymnasium) using motor learning principles for a period of 16 weeks. Outcome measures included the Structured Assessment of Instrumental Living Skills (SAILS), a performance measure with criterion and timed components; a self-report health status questionnaire, the Short Form-36 (SF-36); and the patient-orientated goal directed Canadian Occupational Performance Measure (COPM). RESULTS: There were no group differences on any of the outcome measures: SAILS (p = .3); the SF-36 physical (p = .83) and mental (p = .51); and the COPM performance scale (p = .94) and satisfaction scale (p = .40). CONCLUSIONS: Although we have not excluded benefits of contextually appropriate rehabilitation environments with different intervention approaches, at different stages of rehabilitation or with patients at higher functional levels, our results suggest the appropriateness of a moratorium on these expensive interventions pending demonstration of clear positive effects determined from further study. PMID- 11034231 TI - Worldwide incidence of hip fracture in elderly women: relation to consumption of animal and vegetable foods. AB - BACKGROUND: Hip fracture, a major health problem in elderly persons, varies in incidence among the populations of different countries and is directly related to animal protein intake, a finding that suggests that bone integrity is compromised by endogenous acid production consequent to the metabolism of animal proteins. If that is so, vegetable foods might provide a countervailing effect, because they are a rich source of base (bicarbonate) in the form of metabolizable organic anions, which can neutralize protein-derived acid and supply substrate (carbonate) for bone formation. METHODS: We analyzed reported hip fracture incidence (HFI) data among countries (N = 33) in women aged 50 years and older, in relation to corresponding country-specific data on per capita consumption of vegetable and animal foods as reported by the United Nations Food and Agriculture Organization. RESULTS: HFI varied directly with total (r = +.67, p < .001) and animal (r = +.82, p < .001) protein intake and inversely with vegetable protein intake (r = .37, p < .04). The countries in the lowest tertile of HFI (n = 11) had the lowest animal protein consumption, and invariably, vegetable protein (VP) consumption exceeded the country's corresponding intake of animal protein (AP): VP/AP > 1.0. By contrast, among the countries in the highest tertile of HFI, animal protein intake exceeded vegetable protein intake in nearly every case (10 of 11 countries). Among all countries, HFI correlated inversely and exponentially with the ratio of vegetable/animal protein intake (r = -.84, p < .001) and accounted for 70% of the total variation in HFI. Adjusted for total protein intake, vegetable food consumption was an independent negative predictor of HFI. All findings were similar for the subset of 23 countries whose populations are predominantly Caucasian. CONCLUSION: The findings suggest that the critical determinant of hip fracture risk in relation to the acid-base effects of diet is the net load of acid in the diet, when the intake of both acid and base precursors is considered. Moderation of animal food consumption and an increased ratio of vegetable/animal food consumption may confer a protective effect. PMID- 11034232 TI - Limiting comorbid conditions and breast cancer stage at diagnosis. AB - BACKGROUND: Contrasting hypotheses exist regarding the relationship between comorbidity and breast cancer stage at diagnosis. One suggestion is that disabling comorbid conditions would result in a later stage diagnosis of breast cancer because such conditions would limit mobility and thus access to medical care. This article examines this hypothesis by building a comorbidity summary measure of functionally limiting comorbid conditions and by testing the effectiveness of this measure in predicting the stage at which breast cancer is diagnosed. METHODS: Cases with newly diagnosed breast cancer were identified through the population-based Metropolitan Detroit Cancer Surveillance System, a participant of the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute. Of 1191 eligible cases, 1011 (85%) were interviewed 2 4 months following diagnosis. The analyses for this study were limited to 731 cases for which there were complete data on all variables. Five individual comorbid conditions that predicted functional limitation were combined into a comorbidity summary measure: arthritis, eye conditions, gastrointestinal conditions, kidney conditions, and respiratory conditions. Breast cancer stage was categorized in relation to whether women had local or advanced (regional or remote) disease. RESULTS: Women with two or more of these five functionally limiting conditions were about half as likely as those with none of these conditions to receive an advanced stage diagnosis of breast cancer (odds ratio [OR] = 0.49, 95% confidence interval [CI] 0.28-0.86, p = .01). CONCLUSIONS: These findings do not support the suggestion that the presence of disabling comorbid conditions results in later stage breast cancer. The five conditions summarized by this measure, although functionally limiting, may also require greater medical monitoring due to associated symptoms and/or treatment requirements and thus lead to increased opportunities for cancer screening. PMID- 11034233 TI - Variations in the care of elderly persons with diabetes among endocrinologists, general internists, and geriatricians. AB - BACKGROUND: The American Diabetes Association (ADA) clinical practice recommendations have been widely promoted, but they lack a geriatric-specific approach to care. We aimed to determine the style of care that endocrinologists, general internists, and geriatricians provided to their elderly patients with diabetes and to what extent these medical professionals adhered to the ADA standards. METHODS: We performed a retrospective cohort study of a stratified sample of 531 diabetic patients aged 65 years and older from the endocrinology, general internal medicine, and geriatrics clinics of an urban academic medical center. RESULTS: Patients of geriatricians were older, had higher comorbidity, and were more likely to be demented. The average number of diabetic complications was similar across the specialties, although patients of endocrinologists had higher prevalence of neuropathy and retinopathy compared with patients of geriatricians. Endocrinologists were more likely to use insulin, multiple types of insulin, and combined oral hypoglycemic and insulin therapies. Most patients had hemoglobin A1c measured, and average values were similarly high across specialties at 8.6%. Blood pressures were above 130/85 mm Hg in 85% of the patients. All specialties rarely measured urine microalbumin; geriatricians seldom performed fractionated cholesterol tests, and ophthalmology visits occurred in only half of the patients. CONCLUSION: Endocrinologists had the most aggressive, complex diabetes treatment regimens, although geriatricians had older patients with more dementia and lower prevalence of microvascular complications. Average hemoglobin A1c levels and blood pressures were higher than recommended among patients of all three specialties. Screening for diabetic complications and hyperlipidemia was lower than advised. PMID- 11034234 TI - Determinants of body composition in postmenopausal women. AB - BACKGROUND: Little is known about the effects of different levels of long-term physical activity on total body and regional fat and whether hormone replacement therapy interacts with physical activity level to affect body composition in postmenopausal women. METHODS: We determined the associations between different levels of habitual physical activity, hormone replacement therapy (HRT), and total and regional body composition in postmenopausal women. Twenty sedentary, 20 active nonathletic, and 23 endurance-trained women (approximately half on HRT) had total and regional body composition assessed by dual-energy x-ray absorptiometry. The athletes and active nonathletic women had been active for the same number of years and the same number of hours per week. RESULTS: The athletes and sedentary women weighed the same, but the active nonathletic groups on and not on HRT weighed 3-12 kg more (p < .05). Athletes had less trunk, arm, leg, and total body fat than sedentary and active nonathletic women (p < .05). Women on HRT tended to have lower total body (p = .07), but not regional, fat values. Linear regression analyses indicated that VO2max in ml/kg/min was the major independent determinant of total and regional body fat accounting for 52% to 70% of their variances. Athletes had greater caloric and carbohydrate intake than their less active peers, but all groups had similar protein, fat, saturated fat, monounsaturated fat, and polyunsaturated fat intakes. CONCLUSIONS: Intense training, but not low- to moderate-intensity physical activity, is associated with markedly lower levels of total and regional body fat in postmenopausal women. HRT has less of an effect on body composition than intense exercise training in postmenopausal women. PMID- 11034235 TI - Vitamin and mineral supplement use by older rural adults. AB - BACKGROUND: Vitamin and mineral supplement products are widely consumed by older adults. This study describes supplement product use in a multiethnic rural population, relates supplement usage to dietary nutrient intake, and determines predictors of supplement usage. METHODS: Data are from a population-based sample of 130 community-dwelling adults aged 70 years and older in two rural North Carolina counties. The sample was 34% African American, 36% European American, and 30% Native American. Interviewer-administered semiquantitative food frequency questionnaires were used to obtain data on usual diet and supplement use. In-home interviews allowed verification of supplement composition. Intakes from diet and supplement products were examined for vitamins A, E, B6, C, folate, iron, zinc, and calcium. RESULTS: Of those who participated in the study, 47% reported using one or more supplement products. African Americans were significantly less likely to take supplements than Native Americans or European Americans. Based on dietary intakes, 65% of the participants were deficient (<2/3 recommended dietary allowance [RDA]) for at least one nutrient. The use of supplement products for the eight nutrients investigated was not related to dietary nutrient deficiency. For all nutrients investigated, except iron and calcium, a greater proportion of those without dietary deficiency took a supplement product than those with deficiency. Using logistic regression, ethnicity (European American and Native American), and gender (women) were significant predictors of supplement use. CONCLUSIONS: These findings suggest that although both dietary deficiencies of vitamins and minerals and supplement use are relatively high in this population, there is no association between supplement use and deficient dietary intakes for the eight nutrients examined. Health care providers should be aware that nutritional counseling and guidance on appropriate supplement usage is needed in this population. PMID- 11034236 TI - High urinary catecholamine excretion predicts mortality and functional decline in high-functioning, community-dwelling older persons: MacArthur Studies of Successful Aging. AB - PURPOSE: Catecholamine release is a marker of stress, and high plasma norepinephrine levels have been associated with increased mortality. The predictive value of high urinary catecholamine excretion for functional decline and mortality in healthier older persons has not been determined. SUBJECTS AND METHODS: We used data from the MacArthur Studies of Successful Aging to determine the effects of high urinary catecholamine excretion on 3- and 7-year mortality and functional decline. In 1988, 765 high-functioning older subjects provided complete overnight urine samples for norepinephrine and epinephrine, and 199 of these provided repeat samples in 1991. Subjects who were in the top tertile of urinary norepinephrine or epinephrine excretion in 1988 were considered high excreters; those in the top tertile in both 1988 and 1991 were considered sustained high excreters. We used bivariate and multivariate analysis to examine the relations between high catecholamine excretion and mortality and Rosow Breslau functional decline in 1991 and 1995. RESULTS: In multivariate analyses, subjects with high baseline urinary excretion of epinephrine, norepinephrine, or either catecholamine were at higher risk for mortality and functional decline at 3 and 7 years, although the magnitude of risk (adjusted odds-ratios ranged from 1.1 to 3.1) varied depending upon specific catecholamine and outcome measure. Subjects who had sustained high urinary norepinephrine excretion were also at increased risk for 4-year mortality or functional decline. CONCLUSIONS: High urinary catecholamine excretion in high-functioning, community-dwelling older persons likely reflects subclinical sympathetic stimulation and is a marker of increased risk for functional decline and mortality. PMID- 11034237 TI - Axillary metastases with DCIS: is the glass half empty or half full? PMID- 11034238 TI - Increasing the number of patients undergoing resection of colorectal liver metastases. PMID- 11034239 TI - Sentinel lymph node biopsy: is it indicated in patients with high-risk ductal carcinoma-in-situ and ductal carcinoma-in-situ with microinvasion? AB - BACKGROUND: Axillary lymph node status is the strongest prognostic indicator of survival for women with breast cancer. The purpose of this study was to determine the incidence of sentinel node metastases in patients with high-risk ductal carcinoma-in-situ (DCIS) and DCIS with microinvasion (DCISM). METHODS: From November 1997 to November 1999, all patients who underwent sentinel node biopsy for high-risk DCIS (n = 76) or DCISM (n = 31) were enrolled prospectively in our database. Patients with DCIS were considered high risk and were selected for sentinel lymph node biopsy if there was concern that an invasive component would be identified in the specimen obtained during the definitive surgery. Patients underwent intraoperative mapping that used both blue dye and radionuclide. Excised sentinel nodes were serially sectioned and were examined by hematoxylin and eosin and by immunohistochemistry. RESULTS: Of 76 patients with high-risk DCIS, 9 (12%) had positive sentinel nodes; 7 of 9 patients were positive for micrometastases only. Of 31 patients with DCISM, 3 (10%) had positive sentinel nodes. 2 of 3 were positive for micrometastases only. Six of nine patients with DCIS and three of three with DCISM and positive sentinel nodes had completion axillary dissection; one patient with DCIS had an additional positive node detected by conventional histological analysis. CONCLUSIONS: This study documents a high incidence of lymph node micrometastases as detected by sentinel node biopsy in patients with high-risk DCIS and DCISM. Although the biological significance of breast cancer micrometastases remains unclear at this time, these findings suggest that sentinel node biopsy should be considered in patients with high-risk DCIS and DCISM. PMID- 11034240 TI - Survival after resection of multiple hepatic colorectal metastases. AB - BACKGROUND: Hepatic resection is potentially curative in selected patients with colorectal metastases. It is a widely held practice that multiple colorectal hepatic metastases are not resected, although outcome after removal of four or more metastases is not well defined. METHODS: Patients with four or more colorectal hepatic metastases who submitted to resection were identified from a prospective database. Number of metastases was determined by serial sectioning of the gross specimen at the time of resection. Demographic data, tumor characteristics, complications, and survival were analyzed. RESULTS: From August 1985 to September 1998, 155 patients with four or more metastatic tumors (range 4 20) underwent potentially curative resection by extended hepatectomy (39%), lobectomy (42%), or multiple segmental resections (19%). Operative morbidity and mortality were 26% and 1%, respectively. Actuarial 5-year survival was 23% for the entire group (median = 32 months) and there were 12 actual 5-year survivors. On multivariate analysis, only number of hepatic tumors (P = .005) and the presence of a positive margin (P = .003) were independent predictors of poor survival. CONCLUSIONS: Hepatic resection in patients with four or more colorectal metastases can achieve long-term survival although the results are less favorable as the number of tumors increases. Number of hepatic metastases alone should not be used as a sole contraindication to resection, but it is clear that the majority of patients will not be cured after resection of multiple lesions. PMID- 11034241 TI - Is routine intraoperative frozen-section examination of sentinel lymph nodes in breast cancer worthwhile? AB - BACKGROUND: Routine intraoperative frozen section (FS) of sentinel lymph nodes (SLN) can detect metastatic disease, allowing immediate axillary dissection and avoiding the need for reoperation. Routine FS is also costly, increases operative time, and is subject to false-negative results. We examined the benefit of routine intraoperative FS among the first 1000 patients at Memorial Sloan Kettering Cancer Center who had SLN biopsy for breast cancer. METHODS: We performed SLN biopsy with intraoperative FS in 890 consecutive breast cancer patients, none of whom had a back-up axillary dissection planned in advance. Serial sections and immunohistochemical staining for cytokeratins were performed on all SLN that proved negative on FS. The sensitivity of FS was determined as a function of (1) tumor size and (2) volume of metastatic disease in the SLN, and the benefit of FS was defined as the avoidance of a reoperative axillary dissection. RESULTS: The sensitivity of FS ranged from 40% for patients with Tla to 76% for patients with T2 cancers. The volume of SLN metastasis was highly correlated with tumor size, and FS was far more effective in detecting macrometastatic disease (sensitivity 92%) than micrometastases (sensitivity 17%). The benefit of FS in avoiding reoperative axillary dissection ranged from 4% for Tla (6 of 143) to 38% for T2 (45 of 119) cancers. CONCLUSIONS: In breast cancer patients having SLN biopsy, the failure of routine intraoperative FS is largely the failure to detect micrometastatic disease. The benefit of routine intraoperative FS increases with tumor size. Routine FS may not be indicated in patients with the smallest invasive cancers. PMID- 11034242 TI - Ductal carcinoma-in-situ: long-term results of breast-conserving therapy. AB - BACKGROUND: The role of breast-conserving therapy (BCT) in the management of ductal carcinoma-in-situ (DCIS) is controversial because of reported high recurrence rates. We reviewed our experience to determine whether the rate and pattern of locoregional recurrence after BCT were similar in patients with DCIS and patients with early-stage (T1) invasive breast tumors and whether local recurrence affected survival. METHODS: Between 1973 and 1994, 87 patients with DCIS alone, 22 patients with DCIS with microinvasion (DCIS-M), and 646 patients with invasive breast cancer 2 cm or smaller in diameter were treated with BCT (wide local excision with radiotherapy) at The University of Texas M. D. Anderson Cancer Center. Survival was calculated by the Kaplan-Meier method. The median follow-up times were 11 years for patients with DCIS alone, 12 years for patients with DCIS-M, and 8 years for patients with invasive breast cancer. RESULTS: Eleven (13%) of 87 patients with DCIS and 5 (23%) of 22 patients with DCIS-M had developed locoregional recurrences at follow-up. Two patients with DCIS with locoregional recurrence died of breast cancer. Of the 646 patients with invasive breast cancer, 56 (9%) had a locoregional recurrence, and 16 (2%) died of breast cancer. The median time to locoregional recurrence was significantly longer in patients with DCIS or DCIS-M (9-10 years) than patients with invasive tumors (5 years). CONCLUSIONS: DCIS is a favorable disease with an excellent long-term survival. The locoregional recurrence rate in patients with DCIS treated with BCT is similar to that in patients with early-stage invasive breast cancer treated with BCT, but time to locoregional recurrence is significantly longer in patients with DCIS. In patients with DCIS treated with BCT, intense surveillance for locoregional recurrence needs to be maintained for the patient's lifetime. PMID- 11034243 TI - Postexcision mammography is indicated after resection of ductal carcinoma-in-situ of the breast. AB - BACKGROUND: The adequacy of excision of ductal carcinoma-in-situ (DCIS) usually is confirmed with specimen mammography and histopathological assessment of specimen margins. Postexcision mammography of the involved breast is used at some centers. The objective of this study was to evaluate the impact of postexcision mammography in DCIS. METHODS: We conducted a retrospective chart review of all patients treated for DCIS at our institution from 1995 to 1998. RESULTS: Sixty seven patients had postexcision mammography performed. Residual microcalcifications were identified in 16 patients (24%). Further surgery was precluded by precise mammographic-pathological correlation by using sliced specimen mammography in two patients. Twelve patients had repeat wide excision, and two patients underwent mastectomy. Residual DCIS was identified at re excision in 9 of 14 patients (64%). The margin status of the initial resection was negative in three of nine patients (33%) and positive or unknown in six of nine patients (67%). CONCLUSIONS: Postexcision mammography is a valuable technique that complements specimen mammography and histopathological margin assessment in confirming that an adequate excision of DCIS has been performed. Postexcision mammography should be performed in all patients with DCIS associated with mammographic calcifications who are treated with breast-conserving therapy. PMID- 11034244 TI - Total gastrectomy with dissection of lymph nodes along the splenic artery: a pancreas-preserving method. AB - BACKGROUND: In Japan, the tail and body of the pancreas are generally removed for dissection of lymph nodes along the splenic artery. A new pancreas-preserving method was developed to decrease the postoperative complications due to pancreatic resection. PATIENTS AND METHODS: Between 1981 and 1989, 110 patients were registered in a randomized controlled trial, which included total gastrectomy plus dissection of lymph node along the splenic artery, either with (55 patients: Group A) or without (55: Group B) pancreas tail resection. In Group B, the splenic artery and spleen were removed and the pancreas was preserved. There were no significant differences between the two groups in terms of sex, age, location, microscopic classification, or disease stage. The postoperative complications and survival rates were compared between the two groups. RESULTS: The average number of dissected nodes along the splenic artery was 4.6 and 4.1 for Groups A and B, respectively. The amounts of blood loss during the operation were 994 ml and 904 ml for groups A and B, respectively. Anastomosis failure and/or pancreatic fistula occurred in nine patients in Group A (16%) and seven in Group B (13%). One year after the operation, a glucose tolerance test showed diabetes in 6% and impaired glucose tolerance in 33% of patients in group A, while these findings were normal in group B. The 5-year survival rates were 80% and 76.7% for groups A and B, respectively. CONCLUSIONS: The pancreas-preserving method described here was superior to the more common pancreas resecting method with regard to surgical risk and postoperative glucose tolerance. PMID- 11034246 TI - Prevention of local recurrence after surgical debulking of nodal and subcutaneous melanoma deposits by hypofractionated radiation. AB - BACKGROUND: Local recurrence (LR) after surgical debulking of nodal or subcutaneous melanoma deposits defeats the purpose of operation and may worsen prognosis if the procedure was performed for stage III disease. To decrease LR rates in this setting, we extended the previously described role of hypofractionated radiation for melanoma deposits of the neck to all situations where the patient was felt to be at high risk for postoperative relapse after resection of bulky disease. METHODS: Hypofractionated external beam radiation was administered in 6-Gy doses for 5 fractions (total dose 30 Gy, given over a median of 15 elapsed days) to 42 resected melanoma deposit sites in 41 patients. RESULTS: Stages of the 41 patients at the time of treatment were: 22 stage III and 19 stage IV. All patients had complete gross resection of disease at the radiation site before radiation. Mean time between operation and initiation of radiation was 4 weeks. The 42 sites of treatment included 27 neck, 9 axilla, 3 groin, and 3 subcutaneous deposits. There were no treatment-related deaths; side effects were minimal and self-limited. Transient erythema, desquamation, fibrosis, telangiectasias, and mucositis, parotiditis, and xerostomia (for head and neck radiation) were reported, but no patient required interruption of therapy for these events. Of the 42 treated sites, only 2 recurred in the treatment field (one neck, one axilla) during the mean follow-up time of 22.4 months, for a treatment failure rate of 4.8%. This represents improved local control compared with patients treated with surgery alone at our institution and with published recurrence rates. CONCLUSIONS: The addition of hypofractionated radiation therapy after resection of nodal and subcutaneous melanoma deposits at a variety of sites is a rapid and well-tolerated method of providing excellent local control. PMID- 11034245 TI - A prospective randomized trial comparing intravenous 5-fluorouracil and oral doxifluridine as postoperative adjuvant treatment for advanced rectal cancer. AB - BACKGROUND: Postoperative adjuvant chemoradiation treatment after curative resection for rectal cancer was needed to reduce recurrence and improve a survival rate. Intravenous 5-fluorouracil (5-FU) and leucovorin has been a mainstay of chemotherapy, but oral 5-FU derivatives have been shown a comparable antitumor activity. Intravenous 5-FU and oral doxifluridine were compared with respect to therapeutic efficacy, drug toxicity, and quality of life. METHODS: A total of 166 patients were randomized to receive intravenous 5-FU (450 mg/m2/day) or oral doxifluridine (900 mg/m2/day) in combination with leucovorin (20 mg/m2/day) for depth of invasion, nodal status, metastasis (TNM) stage II and III patients between October 1997 and February 1999. Consecutive daily intravenous infusion for 5 days per every month for a total of 12 cycles (IV arm, n = 74) and oral doxifluridine daily for 3 weeks and 1 week rest for a total of 12 cycles (oral arm, n = 92). Drug toxicity and quality of life were observed. Quality of life was scored according to 22 daily activity items (good, > or =71; fair, < 70; poor, < 52). RESULTS: There was no difference of sex between two groups (IV arm: male/female = 45/29, oral arm: male/female = 59/33). The mean age was 52.3 vs. 59.5, respectively. There was also no difference of TNM stage distribution and type of operation between groups (P>.05). Mean numbers of chemotherapy cycles were 6.5+/-3.7 (IV arm) vs. 7.2+/-4.3 (oral arm), respectively. The rate of recurrence was 9/74 (12.1%) in the IV arm and 6/92 (6.5 %) in the oral arm, respectively (P = .937). Local recurrence was 2/74 (stage III; 2.7%) in the IV arm and 1/92 (stage II; 1.1%) in the oral arm, respectively. Systemic recurrence was 7/74 (stage III; 9.4%) in the IV arm and 5/92 (stage III; 5.4%) in the oral arm, respectively. The most common site of systemic recurrence was the liver. Toxicity profile was as follows: leukopenia (30/74 vs. 17/92) and alopecia (21/74 vs. 13/92) were statistically more common in the IV arm. Diarrhea was more common in the oral arm. Poor quality of life score between two groups was observed at 1 month (23.9% vs. 13%) and 2 months (15.8% vs. 3.7%) after chemotherapy. Good quality of life score was observed at 1 month (19.5% vs. 49%) and 2 months (47% vs. 72%), respectively (P<.05). CONCLUSIONS: Oral doxifluridine with leucovorin shows a comparable therapeutic efficacy to intravenous 5-FU regimen with high quality of life as postoperative adjuvant therapy. The oral regimen also can be safely given with appropriate toxicity and tolerability. PMID- 11034247 TI - Taurolidine inhibits tumor cell growth in vitro and in vivo. AB - BACKGROUND: Taurolidine, a derivative of the amino acid taurine, exhibits antiendotoxin, antibacterial, and antiadherence activity. We hypothesized that Taurolidine may inhibit tumor cell growth, both in an in vitro and in vivo setting. Our aim was to examine the effect of Taurolidine on the growth of a rat metastatic colorectal tumor cell line (DHD/K12/TRb) in vitro and in vivo. METHODS: In the in vitro experiments, DHD/K12/TRb cells were incubated with 5, 10, 15, 25, microg/ml of Taurolidine. Cells incubated in culture medium alone were used as controls. Cell proliferation, cell viability, cell death, and cell apoptosis were measured using commercially available techniques. In the in vivo experiment, BD IX rats were randomized into two groups (n = 10/group). Group A (control) underwent laparotomy and instillation of DHD/K12/TRb tumor cells intraperitoneally followed by phosphate buffered saline (PBS). Group B received Taurolidine (100 mg/kg) instead of PBS. Animals were killed after 24 days and tumor burden assessed by counting the number of tumor nodules in the peritoneal cavity. RESULTS: Incubation of the tumor cells with Taurolidine resulted in a 4 fold decrease in proliferation rates (25+/-4% vs. 100+/-28% for controls) and a 4 fold increase in cell necrosis as demonstrated by the increase in LDH release (403+/-28% vs. 100+/-26% for controls), at a Taurolidine concentration of 25 microg/ml. A dose-dependent decrease in cell viability was also observed. In the in vivo study, local Taurolidine administration resulted in significant decreases in tumor burden (3+/-1 nodules in Group B animals vs. 649+/-101 nodules in Group A animals). CONCLUSIONS: Taurolidine inhibits the growth of a rat metastatic colorectal tumor cell line in vitro and in vivo and thus may have potential in the prevention of peritoneal metastases. PMID- 11034248 TI - Analysis of the genetic alterations in a case of juvenile multiple colon carcinoma with hypogammaglobulinemia. AB - BACKGROUND: We have previously reported the clinical characterization of a case of juvenile multiple colorectal carcinoma with hypogammaglobulinemia. Several recent studies have determined that agammaglobulinemia was caused by the loss of Bruton's tyrosine kinase (Btk) function. However, any genetic alterations associated with carcinoma formation in individuals with this immunodeficient disease have not been reported. METHODS: DNA from eight carcinoma tissues and nine adenoma tissues from this reported case were examined for mutations in p53 by single strand conformation polymorphism analysis, K-ras by mutant allele specific analysis, and replication error or loss of heterozygosity of the TP53 locus on chromosome #17. RESULTS: We found that p53 and K-ras were mutated in the carcinoma tissues. However, each tumor showed unequal and diverse results. CONCLUSIONS: The progression of individual tumor was not due to a common genetic event caused directly under the influence of the primary disease at the genetic level. PMID- 11034249 TI - Dermatofibrosarcoma protuberans of the head and neck. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) of the head and neck is a rare, locally infiltrative, low-grade sarcoma. This study defines the clinical behavior of DFSP, evaluates the role of frozen section analysis, and identifies factors that predict local control. METHODS: Hospital records and pathological slides were reviewed for 33 patients with pathologically confirmed head and neck DFSP treated at Memorial Sloan-Kettering Cancer Center between 1964 and 1999. Factors were analyzed by using Fisher's exact or chi2 tests. RESULTS: For 21 primary and 12 recurrent patients, median age and tumor size at presentation was 39 years and 2.0 cm, respectively. Thirty-two (97%) patients were alive at a median follow-up of 82 months. Three patients recurred locally, all with smaller than 2-cm resection margins. Deep tumors were more likely to have a margin positive resection than superficial lesions (P = .03). Gross margin 2 cm or more was a significant predictor of a negative histological margin (P<.001). There was a trend toward improved recurrence-free survival for tumors treated with wide (> or =2 cm) margin resection (P = .059). Accuracy, sensitivity, specificity, and false negative rates of frozen section were 80%, 43%, 100%, and 57%, respectively. CONCLUSIONS: Wide margin resection of head and neck DFSP predicts negative histological margins and impacts favorably on local recurrence-free survival. Frozen section analysis does not assess resection margins accurately. PMID- 11034250 TI - Gastrointestinal stromal tumors: current diagnosis, biologic behavior, and management. AB - Gastrointestinal stromal tumors (GIST) are rare tumors of the gastrointestinal (GI) tract that arise from primitive mesenchymal cells. GISTs occur throughout the GI tract but are usually located in the stomach and small intestine. The majority of GISTs are immunohistochemically positive for c-kit protein (CD 117) and CD34. GISTs express a heterogeneous clinical course not easily predicted by standard pathological means. The most important prognostic factors are size > 5 cm, tumor necrosis, infiltration and metastasis to other sites, mitotic count > 1 5 per 10 high-powered fields, and most recently, mutation in the c-kit gene. Surgical resection remains the mainstay of treatment, as chemotherapy and radiation are ineffective. Long-term follow-up is imperative, as recurrence rates are high. PMID- 11034251 TI - Surgical therapy of metastatic melanoma. PMID- 11034252 TI - Insurance policies for prophylactic mastectomy: to cover or not to cover? PMID- 11034253 TI - Frequency and clinical outcome of potentially harmful drug metabolic interactions in patients hospitalized on internal and pulmonary medicine wards: focus on warfarin and cisapride. AB - Drug metabolic interactions present potential risks in patient care, but their frequency and relative importance as a clinical problem remains unclear. To assess the frequency and clinical outcome of potentially harmful drug metabolic interactions in hospitalized patients, the authors performed a survey of the medication data of patients treated on internal and pulmonary medicine wards in a university hospital. The database was searched for concomitantly administered drug pairs that would, according to Hansten and Horn's drug interaction database, carry a high risk for a clinically harmful metabolic drug interaction. Coadministrations involving warfarin or cisapride were subjected to further analysis regarding clinical outcome. A total of 142 patients were exposed to 150 interactions with potentially harmful clinical outcome, resulting in a frequency of 0.9% (95% CI 0.7% to 1.0%). Inhibition of warfarin metabolism by metronidazole produced significant overanticoagulation as evidenced by elevated international normalized ratio values, whereas inducers (rifampicin and phenobarbital) of warfarin metabolism significantly reduced the efficacy of warfarin. One case of minor bleeding and one case of clavicular vein thrombosis were detected as possible consequences of disturbed anticoagulation. The coadministration of cisapride and erythromycin significantly prolonged the corrected QT (QTc) interval and was associated with clinical symptoms of cardiac arrhythmias. Coadministration of cisapride with fluconazole or miconazole was not associated with prolongation of the QTc interval or cardiac sequelae. Evaluations of patient materials are needed to assess the clinical relevance of metabolic drug interactions. PMID- 11034254 TI - Genetic polymorphism of CYP2D6 and CYP2C19 metabolism determined by phenotyping Israeli ethnic groups. AB - Genetic polymorphism of the cytochrome P450 isoenzymes CYP2D6 and CYP2C19 was determined by phenotyping four ethnic groups of the Israeli population. The groups consisted of Ethiopian subjects, Yemenite subjects, and Russian subjects representing first-generation new immigrants and an Israeli Arab group. Dextromethorphan was used as the probe for CYP2D6 activity and mephenytoin was used for CYP2C19 activity. The two drugs were administered simultaneously and urine samples were collected over a period of 8 hours. The CYP2D6 phenotype was determined from the ratio of dextromethorphan conversion to dextrorphan and the CYP2C19 phenotype from the ratio of S-mephenytoin and R-mephenytoin. The used liquid chromatographic method was able to completely separate dextrorphan and dextromethorphan. Fluorescence detection allowed dextromethorphan quantification at 1 ng/mL. Mephenytoin enantiomers were completely separated in high-performance liquid chromatography and the respective fractions were collected and analyzed using a gas chromatography/mass spectrometry system with selective ion monitoring. The prevalence of poor metabolizer phenotype of dextromethorphan (CYP2D6) in the Yemenite (0%) and Ethiopian groups (0%) was significantly different from the prevalence in the Russian (17%) and Israeli Arab (9%) groups. A significant difference was also found in the distribution of the metabolic ratio of the extensive metabolizer phenotype between the Ethiopian group and the Russian and Yemenite groups. No significant difference was found in the prevalence of poor mephenytoin metabolizer phenotype (CYP2C19) between the Yemenite (8%), Ethiopian (6%), Russian (9%), and Israeli Arab (8%) groups. No difference was observed in the distribution of metabolic ratio within the extensive metabolizer phenotype subgroups of the four ethnic groups. PMID- 11034255 TI - Saliva and serum concentration of lamotrigine in patients with epilepsy. AB - The authors examined the interindividual correlation between saliva and serum concentrations of lamotrigine (LTG) and the relationship between LTG concentration in saliva and the free LTG concentration in serum in 40 patients with epilepsy, aged 16 to 73 years, receiving stable doses of LTG and comedication. Saliva was collected before and after stimulation of salivary flow. The free LTG fraction was determined by equilibrium dialysis and ultrafiltration. Drug analysis was performed by high-performance liquid chromatography. The correlation between LTG daily dose and serum concentration was weak but significant (r = 0.47). There was high correlation between LTG concentration in serum and unstimulated (r = 0.85) or stimulated (r = 0.94) saliva, and between total LTG concentration in serum and the free LTG fraction as determined by ultrafiltration (r = 0.95) and equilibrium dialysis (r = 0.93). Lamotrigine concentration in stimulated saliva was significantly correlated to the free LTG fraction. Protein binding of LTG calculated from concentration in stimulated saliva, as determined by ultrafiltration and equilibrium dialysis, was 51.8% +/- 13.03%, 68.05% +/- 7.59%, and 58.72% +/- 7.68% (mean +/- standard deviation) respectively. The differences between the three methods were significant. The authors conclude that saliva sampling may be a useful alternative to blood tests for monitoring LTG treatment. PMID- 11034256 TI - Vancomycin pharmacokinetics and Bayesian estimation in pediatric patients. AB - The vancomycin pharmacokinetic profile was characterized in six pediatric patients and the potential of nonlinear mixed effects modeling and Bayesian forecasting for vancomycin monitoring was explored using NONMEM V (1.1). Based on steady state serial vancomycin concentrations, the estimates of mean t1/2, Vd, and Cl derived by the Sawchuk and Zaske method (1) were 3.52 hours, 0.57 L/kg, and 0.12 L/h per kg, respectively. NONMEM analysis demonstrated that a weight adjusted two-compartment model described individual patients' data better than a comparable one-compartment model. The two-compartment estimates of mean t1/2alpha, t1/2beta, Vss, and Cl were 0.80 hour, 5.63 hours, 0.63 L/kg, and 0.11 L/h per kg, respectively. The relatively long mean t1/2alpha suggests that peak vancomycin concentrations measured earlier than 4 hours postdose do not reflect postdistributional serum concentrations. NONMEM population modeling revealed that a weight-adjusted two-compartment model provided a better fit than a comparable one-compartment model. The resulting population parameters and variances were fixed in NONMEM to obtain Bayesian predictions of individual vancomycin serum concentrations. Bayesian estimation with either a single midinterval or trough sample has the potential to provide accurate and precise predictions of vancomycin concentrations. This should be evaluated using a vancomycin population pharmacokinetic model based on a larger sample of pediatric patients. PMID- 11034257 TI - Limited sampling model for the estimation of pharmacokinetic parameters in children. AB - A limited sampling model (LSM) is proposed for the first-time assessment of pharmacokinetic parameters (area under the concentration-time curve (AUC), Cmax, and T1/2) in children after a single oral dose of drug. Three drugs were evaluated in this study. The LSM was developed for each drug from the data of 10 healthy adult volunteers. The relationship at selected time points between plasma concentration and the AUC or Cmax was evaluated by multiple linear regression. The multiple linear regression that gave the best correlation coefficient (r) for 3 sampling times versus AUC or Cmax was chosen as the LSM. Pharmacokinetic parameters generated using sparse sampling (3 blood samples) were compared with pharmacokinetic parameters generated using extensive sampling (>7 blood samples). The results indicated that a limited sampling model can be developed from adult data to estimate pharmacokinetic parameters in children with fair degree of accuracy. PMID- 11034258 TI - Pharmacokinetics and safety of single oral doses of sirolimus (rapamycin) in healthy male volunteers. AB - A phase I study was conducted to determine the pharmacokinetics, safety, and tolerability of sirolimus, a new immunosuppressive drug, in 45 healthy men between 19 and 36 years of age. Nine subjects in each group were randomly assigned to receive single oral doses of either sirolimus (n = 6) or placebo (n = 3) in group I (0.3 mg/m2), group II (1 mg/m2), group III (3 mg/m2), group IV (5 mg/m2) and group V (8 mg/m2). No serious adverse events occurred during the study. Twenty-eight of the 45 volunteers (62%) reported an adverse event; 19 of 30 (63%) were in the sirolimus group and 9 of 15 (60%) were in the placebo group (ns). Asthenia was the most common adverse event, occurring in 7 of 30 (23%) in the sirolimus group compared with 6 of 15 (40%) in the placebo group (ns). Absorption occurred within 1 hour in all volunteers. Whole blood peak concentration and area under the concentration-time curve increased proportionally with dose. Mean (+/- SD) whole blood terminal disposition half life (t1/2), apparent oral dose clearance (Cl/F), and volume of distribution (Vss/F) were 82 +/- 12 hours, 278 +/- 117 mL/h x kg and 23 +/- 10 L/kg, respectively. Distribution of sirolimus into formed blood elements was extensive, with a mean whole blood-to-plasma ratio of 36. Single oral doses of sirolimus (0.3 to 8 mg/m2) solution were well tolerated in healthy male volunteers. PMID- 11034259 TI - Therapeutic drug monitoring in Turkey: experiences from Istanbul. AB - Therapeutic drug monitoring (TDM) has assumed an important place in patient management in the last few decades. In this study, serum drug levels determined in 7759 specimens sent to the Department of Pharmacology and Clinical Pharmacology in 1994 and 1998 for TDM were retrospectively evaluated. Monitored drugs were carbamazepine, valproate, phenytoin, phenobarbital, digoxin, theophylline, and salicylate. The comparison of the results obtained for the relevant 2 years showed that there was a remarkable increase in the number of requests for TDM per year and in the rate of serum drug levels that were within therapeutic range. Serum antiepileptic drug level monitoring accounted for a major part of the data. Overall data suggest that the use of TDM in antiepileptic drugs is improving; conversely, digoxin and theophylline are still not being properly monitored. In this study, the results are discussed in the light of rational TDM criteria. PMID- 11034260 TI - Evaluation of limited sampling strategies for estimation of 12-hour mycophenolic acid area under the plasma concentration-time curve in adult renal transplant patients. AB - Mycophenolate mofetil, the oral prodrug of mycophenolic acid, is indicated as immunosuppressive therapy after renal transplantation. To aid in the investigation of pharmacokinetic-pharmacodynamic relationships of mycophenolic acid in the clinical setting, limited blood sampling strategies have been proposed, and models from these developed, for the estimation of mycophenolic acid area under the concentration-time curve (AUC). In the current study, the authors investigated the predictive performance of six published models to estimate AUC. A total of 49 profiles from 25 renal transplant patients were used to test each model's performance against a full 14 time-point AUC. A wide range of agreement was found when predicted AUCs were compared with full AUCs using linear regression analysis (range: r2 = 0.499 to 0.836). Model 1, which uses 4 time-points over 6 hours, was found to be superior to all other models. The range of time-points used in this model takes into account patients with variable absorption. This model should be further tested on data sets from other centers. The relatively poor performance of the other models may be caused by their inability to describe the peak concentration in these patients. Caution is warranted when using limited sampling strategies on patients whose absorption of mycophenolic acid is altered, compared with those of the pharmacokinetic profiles from which the model was developed. PMID- 11034261 TI - Population pharmacokinetic and pharmacodynamic analysis of ribavirin in patients with chronic hepatitis C. AB - The population pharmacokinetics of ribavirin were assessed in patients with chronic hepatitis C virus (HCV) infection treated with interferon alpha-2b and ribavirin in four clinical efficacy studies. The authors collected 3450 ribavirin serum concentrations from 1105 patients at different treatment weeks for inclusion in the analysis. Population factors included gender, age, body weight, serum creatinine, creatinine clearance, and previous interferon treatment history. Ribavirin apparent clearance (CL/F) was calculated from individual patients' daily doses divided by concentration values, and the influence of these factors was assessed by multiple regression. Body weight, gender, age, and serum creatinine affected CL/F. Population mean CL/F estimates were 17.9 L/h (female) and 21.5 L/h (male) assuming an age of 40 years and body weight of 70 kg. Ribavirin apparent clearance increased as a function of body weight and decreased at ages greater than 40 years. Serum creatinine had little influence on CL/F, which may reflect the relatively normal renal function of these patients. Total CL/F variability was approximately 28%. The four covariates in the model explained 27% of this variability, and were thus of limited clinical significance because of the substantial residual variability not accounted for by the model. In assessing the relationship between pharmacokinetics and pharmacodynamics, the week 4 hemoglobin nadir value was negatively associated with week 4 ribavirin concentrations. The percentage of reduction from baseline was positively associated with ribavirin concentrations, although these data were highly variable. Loss of HCV-RNA at 24 weeks after completion of treatment was considered a response to interferon and ribavirin treatment in a logistic regression analysis of clinical and pharmacokinetic variables and treatment response in the interferon-naive patients. Hepatitis C virus genotype, pretreatment HCV-RNA titer, duration of treatment period, week 4 ribavirin concentration, and patient age affected the likelihood of response. Higher ribavirin concentrations at treatment week 4 were associated with a higher response rate. Variables that have predictive value for treatment outcome in patients treated with interferon and ribavirin are similar to those previously reported for interferon monotherapy. PMID- 11034262 TI - Single dose pharmacokinetics of syrup of ipecac. AB - Syrup of ipecac (SI) has been used medicinally since the 1500s; however, little is known about the pharmacokinetics in humans of SI's active ingredients, emetine and cephaeline. The objective of this study was to evaluate the rate of absorption and the rate of elimination of emetine and cephaeline. Ten healthy, adult, human volunteers between 18 and 45 years of age who were of ideal body weight (body mass index 20-25) completed this study. After an overnight fast, 30 mL SI were ingested. Blood samples were collected 30, 45, 60, 90, 120, 150, and 180 minutes post-ingestion and urine was collected throughout the study period. Plasma and urine concentrations of emetine and cephaeline were measured by reverse-phase HPLC with fluorescence detection. In virtually all subjects, emetine and cephaeline were detected within 5-10 minutes of dosing with the time to maximum concentration being approximately 20 minutes. The mean areas under the concentration-time curve (AUC) for both emetine and cephaeline were similar; however, the ratio of mean cephaeline maximum concentration (Cmax) to emetine Cmax was approximately 1.5. Four of the ten subjects exhibited a type of concentration-time profile in which the levels of cephaeline were substantially higher than those of emetine and the levels of cephaeline were substantially higher than noted for the other six subjects. In these remaining six subjects, the levels of emetine and cephaeline were lower than 10 ng/mL at all time-points. An initial elimination phase was noted in some subjects but not in others. Individuals in whom an initial elimination phase was not observed also exhibited low levels of both alkaloids as compared with the other subjects suggestive of a slower distribution phase. Less than 0. 15% of the administered emetine and cephaeline was recovered in the urine at 3 hours. No relationship between vomiting episodes and peak concentrations of emetine or cephaeline was found. Administration of SI results in rapid appearance and disappearance of emetine and cephaeline in plasma becoming almost undetectable at 3 hours. Very little of either alkaloid is eliminated in the urine within this time period, suggesting extensive distribution. The length of time that an administered dose of SI can result in the detection of emetine and/or cephaeline in the urine has not been determined; future studies in humans are required. PMID- 11034263 TI - Stereoselective pharmacokinetic analysis of valnoctamide, a CNS-active chiral amide analogue of valproic acid, in dogs, rats, and mice. AB - The purpose of this study was to evaluate the stereoselective pharmacokinetics of valnoctamide (VCD) in dogs, rats, and mice; which are the most common animal models for pharmacokinetic, pharmacologic, and toxicologic evaluation; and to compare it with previously published human data. Racemic VCD (mixture of four stereoisomers) was administered intravenously to six mongrel dogs and to rats (five rats per time-point), and intraperitoneally to mice (five mice per time point). Plasma concentrations of the individual stereoisomers were measured by a stereospecific gas chromatography assay. In dogs, (2S,3R)-VCD had a larger clearance (0.33 L/h x kg) and a larger volume of distribution (0.79 L/kg) than its two diastereomers (0.24-0.25 L/h x kg and 0.65 L/kg, respectively). A tendency toward slightly higher clearance and volume of distribution values for (2S,3R)-VCD was observed in rats and mice as well. Consequently, in all three animal species the half-life (t1/2) of (2S,3R)-VCD was not different from the t1/2 of the other three VCD stereoisomers. The stereoselective pharmacokinetics of VCD as observed in dogs, rats, and mice is in line with the stereoselectivity previously observed in healthy subjects and epileptic patients. PMID- 11034264 TI - Digoxin, flecainide, and amiodarone transfer across the placenta and the effects of an elevated umbilical venous pressure on the transfer rate. AB - Clinical observations suggest that flecainide might pass the placenta more easily than digoxin, and that its transfer is less disturbed in case of hydrops fetalis than that of digoxin. The purpose of the study was to compare the materno-fetal transplacental transfer of digoxin, flecainide, and amiodarone, another antiarrhythmic agent used in the treatment of fetal tachyarrhythmia, and to assess the effect of an elevated umbilical venous pressure (UVP) on the transfer rate. Isolated lobules of 16 human placentas were dually perfused after spontaneous delivery or caesarean section. The transplacental transfer (area under the curve in the maternal compartment [maternal AUC], area under the curve in the fetal compartment [fetal AUC], kinetic parameters) of digoxin, flecainide, and amiodarone was calculated after these drugs were added to the maternal circuit. In five experiments, the effect of increased UVP on the transplacental transfer rate was assessed by elevating the UVP by 10 cm H2O. Flecainide efflux out of the maternal compartment was significantly greater than that of digoxin (maternal AUC 57.4% +/- 5.1 %/min vs 73.9% +/- 1.5%/min), whereas the flecainide influx into the fetal circulation was smaller (fetal AUC 9.3% +/- 4.1%/min vs 11.5% +/- 2.0%/min). Only in 50% of the experiments were the smallest amounts of amiodarone detectable in the fetal compartment. An elevation of the UVP reduced the influx of digoxin and flecainide into the fetal compartment (fetal AUC) from 11.5% +/- 2.0%/min to 7.4% +/- 1.9%/min and from 9.3% +/- 4.1% to 4.7% +/- 1.4%/min, respectively. Materno-fetal transplacental transfer of digoxin, flecainide, and amiodarone decreases in this sequence. Fetal cardiac insufficiency accompanied by an elevation of the UVP might reduce the transplacental transfer of these drugs, although no significant difference could be found between the reduction of transfer of digoxin and flecainide. PMID- 11034265 TI - High-performance liquid chromatographic method for the determination of colistin in serum. AB - Inhalation therapy of colistin is widespread in patients with cystic fibrosis. To date, no pharmacokinetic data of colistin after inhalation are available. To optimize the inhalation therapy, pharmacokinetic data of colistin are necessary. In this study, the authors describe a chromatographic analysis for measurement of colistin concentrations in serum. After protein precipitation, the colistin sample is treated with orthophthalaldehyde for derivatization. The sum of the peak areas of the two main components of colistin (polymyxin E1 and E2) were used for quantitation. The performance of the analytical method was assessed by determining the lower limit of quantitation, the selectivity of the method, the intra-assay variation, the reproducibility, the interassay variation, and the accuracy. The lower limit of quantitation was 28 microg/L. Ceftazidime, aztreonam, piperacilline, or tobramycin showed no interference with the colistin assay. In a pilot study, the authors found a trough value of approximately 10 microg/L and peak values of approximately 100 microg/L after inhalation of 160 mg colistin in serum samples of a representative patient. These values show that the method can be used to design further experiments. The applicability of the method was also tested on urine and sputum samples. Colistin was detectable but further validation experiments are required to confirm the usefulness of the method in these biologic matrices. To the authors' knowledge this is the first study in which serum concentrations are described after inhalation of colistin in patients with cystic fibrosis. PMID- 11034266 TI - Accuracy of cyclosporin measurements made in capillary blood samples obtained by skin puncture. AB - International consensus guidelines suggest that cyclosporin should be measured in whole blood. In some instances it may be advantageous to collect capillary blood, by a finger or ear prick method. However, drug concentrations in skin-puncture blood may not necessarily correlate with those measured in venous blood. This study compared cyclosporin concentrations in blood collected from the fingertip or earlobe with blood collected by standard venipuncture. Patient preference for each of the blood collection methods was also assessed. Specimens were obtained from organ transplant patients receiving cyclosporin, using each of the three methods: venipuncture, finger prick, and earlobe prick. The samples were assayed using a specific radioimmunoassay and the results were compared. In the 102 sets of samples collected, the mean difference (+/- standard deviation) in cyclosporin concentration between finger prick and venipuncture and ear prick and venipuncture was 2.6% (+/- 9.5%) and 2.7% (+/- 12.1%), respectively, while the comparable median (IQR) differences were 1.9% (-3.4% to +6.6%) and -1.1% (-2.8% to +7.2%), respectively. A high degree of correlation was observed between finger prick and venipuncture or ear prick and venipuncture or ear prick and finger prick (r2 > 0.86). Of the three methods of blood collection, finger prick was the patients' preferred method (P < 0.01). These data suggest that capillary blood collected by skin puncture is suitable for use in cyclosporin blood monitoring and acceptable to patients. PMID- 11034267 TI - Comparison of the pharmacokinetics and in vivo bioaffinity of DigiTAb versus Digibind. AB - The in vivo digoxin binding affinity and normal pharmacokinetic values of digoxin immune Fab are unknown. Healthy subjects (n = 16) were randomized to one of the two digoxin-immune Fab products, DigiTAb or Digibind, to compare the in vivo digoxin binding affinity and pharmacokinetic disposition. Each subject received 1 mg of intravenous digoxin infused during 5 minutes followed 2 hours later by 76 mg of either DigiTAb or Digibind. Both Fab products reduced free digoxin serum concentrations to below assay detection with equal ability. Consequently, total digoxin serum concentrations increased approximately 10-fold. Peak total digoxin serum concentrations post-Fab dosing were similar to the pre-Fab peak digoxin concentration for both Fab products (45 +/- 14 and 44 +/- 11 for DigiTAb, pre and post, respectively) 50 +/- 17 and 41 +/- 9 for Digibind, pre and post, respectively) indicating in vivo equimolar binding affinity. While bioaffinity for digoxin was equal between groups, total digoxin area under the curve (AUC) and digoxin-immune Fab AUC were lower in the DigiTAb group compared with the Digibind group. Hence, systemic total digoxin and Fab clearance were greater in the DigiTAb-treated group. In conclusion, equimolar doses of both DigiTAb and Digibind completely bind digoxin in vivo. The ability of digoxinimmune Fab to bind to digoxin is not affected by the systemic disposition of the Fab product. PMID- 11034268 TI - Simultaneous quantification of tacrolimus and sirolimus, in human blood, by high performance liquid chromatography-tandem mass spectrometry. AB - In this paper the authors present a validated method for the simultaneous analysis of tacrolimus and sirolimus in human blood by high-performance liquid chromatography-electrospray tandem mass spectrometry. Blood samples (500 microL) were prepared by C18 solid-phase extraction. Mass spectrometric detection was by selected reaction monitoring. The assay was linear for both compounds over the range 0.25-100 microg/L (r2 > 0.996, n = 7). At the limit of quantification (0.25 microg/L), for both sirolimus and tacrolimus, the interday imprecision was < 3% and the analytical recovery was between 97.0% and 102%, respectively. The interbatch and intrabatch coefficients of variation of the method for both analytes, at the three quality control concentrations (0.5, 20, and 80 microg/L), were < 16% and < 10%, respectively. The analytical recovery, at the three control concentrations, ranged from 99.2% to 104% of the nominal concentration. The mean absolute recovery (+/- standard deviation) of tacrolimus, sirolimus, and internal standard was 82 +/- 7%, 89 +/- 12%, and 77 +/- 8%, respectively (n = 12). In conclusion, the method presented can be used for simultaneous determination of tacrolimus and sirolimus and will aid in pharmacokinetic studies and therapeutic drug monitoring of these drugs. Furthermore, this method has economic benefits in the clinical setting where these drugs are coadministered. PMID- 11034269 TI - Determination of ifosfamide, 2- and 3-dechloroethyifosfamide using gas chromatography with nitrogen-phosphorus or mass spectrometry detection. AB - A comparison was made between methods for determining ifosfamide (IF), 2- (2DCE) and 3-dechloroethylifosfamide (3DCE) using gas chromatography with nitrogen phosphorus detection (GC-NPD) versus positive ion electron-impact ion-trap mass spectrometry (GC-MS2). Sample pretreatment involved liquid-liquid extraction with ethyl acetate after adding trofosfamide as internal standard and alkalinization. The GC-NPD was linear, specific, and sensitive for all analytes in the range of 0.0500-100 microg/mL with lower limits of quantification (LLQ) of 0.0500 microg/mL using a 50-microgL plasma sample. The GC-MS2 was linear, specific, and sensitive for IF, 2DCE, and 3DCE in the ranges of 0.250-100, 0.500-25.0, and 0.500-25.0 microg/mL, respectively, with LLQs of 0.250, 0.500, and 0.500 microg/mL. The ranges of accuracy, within-day precision, and between-day precision for analysis of all compounds with GC-NPD did not exceed 93.3% to 105.4%, 8.0% and 9.8%, respectively. The ranges of accuracy, within-day precision, and between-day precision for analysis of all compounds with GC-MS2 did not exceed 86.5% to 99.0%, 9.0% and 12.7%, respectively. In conclusion, GC NPD proved to be superior to GC-MS2 in sensitivity, detection range, accuracy, and precisions. Therefore GC-NPD is the method of choice for fast un-derivatized determination of IF, 2DCE, and 3DCE in human plasma, and it can readily be used for clinical pharmacokinetic studies and routine monitoring of IF-treated patients in a hospital setting. PMID- 11034270 TI - Optimized procedure for lamotrigine analysis in serum by high-performance liquid chromatography without interferences from other frequently coadministered anticonvulsants. AB - The authors have developed a simple isocratic high-pressure liquid chromatographic (HPLC) assay for the simultaneous determination of lamotrigine and other frequently coadministered antiepileptic drugs in serum samples. Lamotrigine extraction was performed on a reversed-phase Oasis HBL preparation column. The eluates containing butalbital as internal standard were separated with a 7-microm Chromsystems C18 250 x 4.0 mm I.D. reversed-phase column at a temperature of 40 degrees C using a mobile phase consisting of pH 3.8 phosphate acetonitrile buffer (55:45, v/v), at a flow rate of 0.8 mL/min. Ultraviolet detection was carried out at 210 nm. Measurement of the peak:height ratio allowed quantitative determination of the samples. The method was linear over a concentration range of 0.2 to 20 microg/mL for lamotrigine. Recovery was >90%. Within-day and between-day coefficients of variation ranged from 1.8% to 6.7%. The mean lamotrigine concentration was 8.01 +/- 5.63 microg/mL. After studying sera from 130 patients treated with lamotrigine the authors confirmed that associated antiepileptic therapy affected the serum lamotrigine levels, which were significantly higher in patients under valproic acid treatment. PMID- 11034271 TI - Rapid and simple determination of indinavir in serum, urine, and cerebrospinal fluid using high-performance liquid chromatography. AB - A method for analysis of indinavir in serum, cerebrospinal fluid, and urine was developed. The method is based on liquid-liquid extraction followed by high performance liquid chromatography with UV detection. The method has a shorter analysis time than previously published methods, and it is sensitive enough to measure levels in all three fluids under routine clinical conditions. The method is linear up to 32 micromol/L, the limit of detection is 0.01 micromol/L, and recovery of the method is 86%. The interassay coefficient of variation at 2.0 micromol/L was 2.8%, and no internal standard is needed. Over 700 clinical samples have been analyzed by this method, and concomitant antiviral drugs do not interfere with the assay. Paroxetin and dipyridamol are the only two compounds encountered to elute with retention times similar to that of indinavir. Examples of chromatograms and a pharmacokinetic curve are given. The method is well suited for routine therapeutic drug monitoring as well as for pharmacokinetic studies for research purposes. PMID- 11034272 TI - Cocaine and cocaethylene binding to normal human brain and Alzheimer disease brain. AB - The binding of cocaine and cocaethylene to homogenates of both normal whole brain and whole brain with Alzheimer disease patients (Alzheimer brain) was investigated in vitro using equilibrium dialysis of the unlabelled drugs at 4 degrees C. Two binders of cocaine were characterized in normal brain (binder 1: Ka, 5.73 x 10(3) L/mol; Bo, 7.44 x 10(-5) mol/L) (binder 2: Ka, 1.54 x 10(3) L/mol; Bo, 2.50 x 10(-4) mol/L) and in Alzheimer brain (binder 1: Ka, 3.08 x 10(2) L/mol; Bo, 6.66 x 10(-4) mol/L) (binder 2: Ka, 8.74 x 10(1) L/mol; Bo, 4.30 x 10(-3) mol/L). For cocaethylene three binders were noted in normal brain (binder 1: Ka, 3.23 x 10(3) L/mol; Bo, 1.22 x 10(-4) mol/L) (binder 2: Ka, 3.10 x 10(3) L/mol; Bo, 2.01 x 10(-4) mol/L) (binder 3: Ka, 1.63 x 10(3) L/mol; Bo, 3.59 x 10(-4) mol/L) and two binders in Alzheimer brain (binder 1: Ka, 5.18 x 10(3) L/mol; Bo, 3.06 x 10(-5) mol/L) (binder 2: Ka, 3.36 x 10(3) L/mol; Bo, 7.75 x 10( 5) mol/L). The binding of cocaine to normal brain was much stronger than to Alzheimer brain (ten- to 100-fold), whereas the binding of cocaethylene was similar in normal and Alzheimer brain. Cocaine and cocaethylene binding to human brain was compared with cocaine and cocaethylene binding to other human tissues previously studied by this laboratory. PMID- 11034273 TI - Fluconazole-induced convulsions at serum trough concentrations of approximately 80 microg/mL. AB - On the basis of two case reports it is suggested that serious convulsions may occur in patients treated with flucoazole when serum trough concentrations exceed 80 microg/mL. The authors recommend monitoring fluconazole concentrations during high-dose therapy in patients with poor kidney function. PMID- 11034274 TI - Switching antiretroviral therapy from an enzyme-inducing drug. PMID- 11034275 TI - Drug interaction between mycophenolate mofetil and tacrolimus detectable within therapeutic mycophenolic acid monitoring in renal transplant patients. PMID- 11034276 TI - Treatment options for extended-spectrum beta-lactamase-producers. AB - A review of antibiotic options for the treatment of infections caused by extended spectrum beta-lactamase-producing isolates is presented. The use of the third generation cephalosporin, cefotaxime, for infections caused by isolates producing ceftazidimase-type extended-spectrum beta-lactamases is controversial, despite in vitro susceptibility to the antibiotic in many instances. The fourth-generation cephalosporin, cefipime, although active against most extended-spectrum beta lactamases, is reported to show a marked inoculum effect. The cephamycins, such as cefoxitin. are generally effective against Enterobacteriaceae producing TEM- and SHV-derived extended-spectrum beta-lactamases, but Klebsella pneumoniae strains are prone to cephamycin resistance as a result of porin loss. The use of beta-lactamase inhibitor combinations is variable. Sulbactam is less effective than clavulanate for the inhibition of SHV-derived extended-spectrum beta lactamases and a marked inoculum effect has been noted, while the efficacy of tazobactam against SHV-derived extended-spectrum beta-lactamase producers is controversial. Furthermore, extended-spectrum beta-lactamases are often encoded by multi-resistant plasmids carrying genes conferring resistance to aminoglycosides, chloramphenicol, sulfonamides, trimethoprim and other antimicrobials, severely limiting even alternative therapies. Extensive susceptibility testing before the institution of antibiotic therapy is thus vital. PMID- 11034277 TI - A novel degradative pathway of 2-nitrobenzoate via 3-hydroxyanthranilate in Pseudomonas fluorescens strain KU-7. AB - A bacterial strain KU-7, identified as a Pseudomonas fluorescens by 16S rDNA sequencing, was one of the 12 new isolates that are able to grow on 2 nitrobenzoate as a sole source of carbon, nitrogen, and energy. Resting cells of KU-7 were found to accumulate ammonia in the medium indicating that degradation of 2-NBA proceeds through a reductive route. Metabolite analyses by thin layer chromatography and high pressure liquid chromatography indicated that 3 hydroxyanthranilate is an intermediate of 2-nitrobenzoate metabolism in KU-7 cells. This offers an alternative route to 2-nitrobenzoate metabolism since anthranilate (2-aminobenzoate) or catechol were detected as intermediates in other bacteria. Crude extracts of KU-7 cells converted 2-nitrobenzoate to 3 hydroxyanthranilate with oxidation of 2 mol of NADPH. Ring cleavage of 3 hydroxyanthranilate produced a transient yellow product, identified as 2-amino-3 carboxymuconic 6-semialdehyde, that has a maximum absorbance at 360 nm. The initial enzymes of the 2-nitrobenzoate degradation pathway were found to be inducible since succinate-grown cells produced very low enzyme activities. A pathway for 2-nitrobenzoate degradation in KU-7 was proposed. PMID- 11034278 TI - Binding of catalase by Gardnerella vaginalis. AB - Previous work has demonstrated that Gardnerella vaginalis can utilize catalase as a sole source of iron. In this study, the interaction between G. vaginalis cells and catalase was investigated. G. vaginalis cells were shown to bind digoxigenin (DIG)-labeled catalase using a solid phase dot blot assay. An increase in catalase binding was observed from cells grown under iron-restrictive conditions. Western blot analysis of G. vaginalis proteins resulted in the detection of a putative catalase-binding protein with an estimated molecular mass of 128 kDa. The 128-kDa catalase-binding protein was not detected from intact G. vaginalis cells treated with trypsin prior to Western blot analysis suggesting this protein may be surface-exposed. PMID- 11034279 TI - Characterization of the relA/spoT gene from Bacillus stearothermophilus. AB - By use of degenerate primers, we amplified a fragment of a relA/spoT homologous gene from Bacillus stearothermophilus. Chromosomal walking enabled us to sequence the entire gene and its flanking regions. The primary sequence of the gene product is 78% identical to the RelA/SpoT homologue of Bacillus subtilis and both gene loci share a similar genetic organization. The B. stearothermophilus rel gene was analyzed in vivo by heterologous expression in the B. subtilis relA deletion strain TW30, and is shown to complement the growth defects of TW30. The recombinant RelBst protein was detected by Western immunoanalysis, and synthesizes guanosine-3'-diphosphate-5'-(tri)diphosphate ((p)ppGpp) after amino acid stress and carbon starvation. These in vivo data, the genetic organization, and the primary structure compared to other RelA/SpoT homologues provide circumstantial evidence that the identified gene encodes the only (p)ppGpp synthetase in B. stearothermophilus presumed to serve also as (p)ppGpp hydrolase. PMID- 11034280 TI - Molybdate-dependent expression of dimethylsulfoxide reductase in Rhodobacter capsulatus. AB - Expression of the dimethylsulfoxide respiratory (dor) operon of Rhodobacter is regulated by oxygen, light intensity and availability of substrate. Since dimethylsulfoxide reductase contains a pterin molybdenum cofactor, the role of molybdate in the regulation of dor operon expression was investigated. In this report we show that the molybdate-responsive transcriptional regulator, MopB, and molybdate are essential for maximal dimethylsulfoxide reductase activity and expression of a dorA::lacZ transcriptional fusion in Rhodobacter capsulatus. In contrast, mop genes are not required for the expression of the periplasmic nitrate reductase or xanthine dehydrogenase in R. capsulatus under conditions of molybdenum sufficiency. This is the first report demonstrating a clear functional difference between the ModE homologues MopB and MopA in this bacterium. The results suggest that MopA is primarily involved in the regulation of nitrogen fixation gene expression in response to molybdate while MopB has a role in nitrogen fixation and dimethylsulfoxide respiration. PMID- 11034281 TI - Expression of a cDNA encoding a Toxoplasma gondii protein belonging to the heat shock 90 family and analysis of its antigenicity. AB - A cDNA clone (Tgzy85d11.r1) obtained from the Toxoplasma Expressed Sequence Tag project was chosen due to its homology with proteins of the heat shock 90 family. The cDNA encodes 137 amino acids of the C-terminal portion of the Toxoplasma Hsp90 protein (TgHsp90). Serum samples obtained from orally infected BALB/c and C57BL/6 mice showed reactivity against a recombinant TgHsp90 (rTgHsp90) after 8 weeks postinfection. Isotype analysis showed an anti-rTgHsp90 IgG2a/IgG3 response in infected BALB/c and anti-rTgHsp90 IgG1/IgG2a/IgG2b response in infected C57BL/6 mice. Serum samples from individuals chronically and putative acutely infected with T. gondii showed a similar anti-rTgHsp90 IgG response. Our work identifies TgHsp90 as a novel parasite antigen that seems to elicit a higher relation of anti-TgHsp90/anti-T. gondii IgGs during chronic infection in comparison with the acute stage. PMID- 11034282 TI - Characterization of strain HY99, a novel microorganism capable of aerobic and anaerobic degradation of aniline. AB - We have characterized a novel microorganism, strain HY99, which is capable of aerobic and anaerobic degradation of aniline. Strain HY99 was found to aerobically metabolize aniline via catechol and 2-hydroxymuconic semialdehyde intermediates, and to transform aniline via p-aminobenzoate in anaerobic environments. Physiological and biochemical tests revealed that strain HY99 was most similar to Delftia acidovorans, but unlike D. acidovorans, strain HY99 was able to metabolize aniline under anaerobic conditions linked with nitrate reduction. Phylogenetic analysis based on 16S rDNA sequencing also revealed that strain HY99 was closely related to D. acidovorans, with 96% overall similarity. PMID- 11034283 TI - Cloning, expression and biochemical characterisation of a unique thermostable pullulan-hydrolysing enzyme from the hyperthermophilic archaeon Thermococcus aggregans. AB - The gene for a new type of pullulan hydrolase from the hyperthermophilic archaeon Thermococcus aggregans was cloned and expressed in Escherichia coli. The 2181-bp open reading frame encodes a protein of 727 amino acids. A hypothetical membrane linker region was found to be cleaved during processing in E. coli. The recombinant enzyme was purified 70-fold by heat treatment, affinity and anion exchange chromatography. Optimal activity was detected at 95 degrees C at a broad pH range from 3.5 to 8.5 with an optimum at pH 6.5. More than 35% of enzymatic activity was detected even at 120 degrees C. The enzyme was stable at 90 degrees C for several hours and exhibited a half-life of 2.5 h at 100 degrees C. Unlike all pullulan-hydrolysing enzymes described to date, the enzyme is able to attack alpha-1,6- as well as alpha-1,4-glycosidic linkages in pullulan leading to the formation of a mixture of maltotriose, panose, maltose and glucose. The enzyme is also able to degrade starch, amylose and amylopectin forming maltotriose and maltose as main products. PMID- 11034284 TI - Allelic variation in a peptide-inducible two-component system of Streptococcus pneumoniae. AB - The peptide SpiP of Streptococcus pneumoniae regulates the induction of a complex signal transduction system spiR1spiR2spiH. Distinct alleles of spiP and the receptor histidine protein kinase gene spiH were recognized in different pneumococcal clones. The spi system in strain KNR7/87 is adjacent to a bacteriocin gene cluster encoding putative double glycine-type bacteriocins, immunity proteins, and translocator proteins. A direct repeat element upstream of the spiR1 promoter and another three potential transcription start sites within the bacteriocin cluster indicate that SpiP functions as an inducing peptide for bacteriocin synthesis in S. pneumoniae. PMID- 11034285 TI - Novel sucrose transposons from plant strains of Lactococcus lactis. AB - Lactococcus lactis strains isolated from vegetable products transferred the ability to ferment sucrose in conjugation experiments with the recipient strain L. lactis MG1614. Nisin production and sucrose fermentation were transferred together from two strains, but transfer also occurred from several other strains which did not produce nisin. Pulsed-field gel electrophoresis analysis showed that all transconjugants had acquired large chromosomal insertions at two main sites. Nisin sucrose transconjugants had gained inserts of 70 kb, while those that fermented sucrose without nisin production contained inserts of between 50 and 110 kb. Transconjugants from one donor had acquired a separate insertion of 55 kb which correlated with enhanced bacteriophage resistance, but contained neither nisin nor sucrose fermentation genes. PMID- 11034286 TI - Physiological analysis on novel coculture of Monascus sp. J101 with Saccharomyces cerevisiae. AB - During the fermentation process of Monascus 3101, coculture with Saccharomyces cerevisiae culture filtrate stimulated the formation of reproductive spores, which subsequently resulted in accelerated Monascus cell reproduction and proliferation. Protein kinase C activity was also detected. Chitinase (EC 3.2.1.14), a 120-kDa secretory protein, was purified from the S. cerevisiae culture filtrate as the effector. Monascus cells cocultured with a S. cerevisiae culture filtrate contained approximately four times more total lipids (mainly linoleic and oleic acid) than Monascus cells without coculture. Addition of exogenous fatty acids only contributed to an increase in cell mass. There was no effect on spore formation or pigment production. There were significant changes in patterns and amounts of expressed proteins in cocultured Monascus cells compared to control cells with no coculture. PMID- 11034287 TI - Characterization of the chemotaxis fli Y and che A genes in Bacillus cereus. AB - This paper describes the first identification of chemotaxis genes in Bacillus cereus. We sequenced and studied the genomic organization and the expression of the cheA and fliY genes in two different B. cereus strains, ATCC 14579 and ATCC 10987. While cheA encodes a highly conserved protein acting as the main regulator of the chemotactic response in flagellated eubacteria, fliY, which has been previously described only in B. subtilis, is one of the three genes encoding proteins of the flagellar switch complex. Although the sequences and relative position of cheA and fliY were found to be identical in the two B. cereus strains analyzed, the restriction fragment containing both genes was located differently on the physical maps of B. cereus ATCC 14579 and ATCC 10987. Evidence is shown that the genomic organization and the expression of fliY and cheA in B. cereus differ significantly from that described for B. subtilis, which is considered a model microorganism for chemotaxis in gram-positive bacteria. PMID- 11034288 TI - Impact of synthetic pyrethroid-sheep dip on the indigenous microflora of animal slurries. AB - The chemical constituents of sheep dip in the UK are currently changing from organophosphate-based to synthetic pyrethroid-based insecticides. As a result, changes are also being made to the methods of disposal of these chemicals in the environment, such that pyrethroid sheep dips must now be diluted in animal slurry or water. To date, there is a lack of quantitative information on the impact of the insecticide on the indigenous microflora of animal slurries. This paper investigated the impact of Bayticol (synthetic pyrethroid sheep dip) over a range of concentrations on selected populations of bacteria within animal slurry. It was found that, with increasing pesticide concentration, there was up to a four orders of magnitude increase in the numbers of faecal coliforms and pathogens, such as putative Salmonella spp. These findings have implications for the disposal of sheep dip-amended animal slurries to land from several aspects: (i) the longevity of putative pathogens in the field may require re-evaluation of the time required before the return of grazing livestock to a slurry-amended field; (ii) the potential for the transfer of pathogenic bacteria and faecal coliforms into human and animal foodchains, and (iii) the increased potential for faecal coliforms being washed into streams, rivers and coastal bathing waters. PMID- 11034289 TI - Ion channel formation by N-terminal domain: a common feature of OprFs of Pseudomonas and OmpA of Escherichia coli. AB - The proteolytic fragments of OprFs of Pseudomonas aeruginosa and Pseudomonas fluorescens were identified, respectively, as the first 175 and 177 amino acids from the N-terminal domain. They induced ion channels after reincorporation into planar lipid bilayers (85 and 75 pS, respectively, in 1 M NaCl). A similar conductance value (72 pS) was found for the eight beta-strand OmpA N-terminal domain (OmpA171) of Escherichia coli. We conclude that the N-terminal domain of OprFs is sufficient to induce ion channels and the comparison with OmpA171, provides strong evidence of the existence of an eight-stranded beta-barrel in the N-terminal domain of OprFs. PMID- 11034290 TI - Changes in the virulence of Mycobacterium avium after passage through embryonated hens' eggs. AB - Eight-day-old embryonated hen's eggs were used as a model to study Mycobacterium avium virulence. Strains isolated from human patients caused 20-90% mortality when eggs were infected by injection of bacterial suspensions into the amniotic sac. Virulence of examined strains subsequently decreased with passage through eggs to between 0 and 40% mortality in four passages. Virulence of the egg attenuated strains could be restored by passage through human peripheral blood mononuclear cells. The site of infection in the egg was usually the mesodermal layer of the chorioallantoic membrane. A few small granulomas containing acid fast bacteria were seen in the liver, but not in other organs. Death of chicken embryos may have resulted from destruction of the mesodermal layer of the chorioallantoic membrane with consequent respiratory failure. PBMCs infected with less virulent egg-passaged strains of M. avium produced higher levels of tumor necrosis factor-alpha than did peripheral blood mononuclear cells infected with more virulent nonpassaged strains. PMID- 11034292 TI - Involvement of naphthalene dioxygenase in indole-3-acetic acid biosynthesis by Pseudomonas putida. AB - Two variants of plant growth-promoting strain Pseudomonas putida BS1380 harboring the naphthalene degradative plasmid pBS2 and the recombinant plasmid pNAU64 that contains the genes encoding for naphthalene dioxygenase were constructed by conjugation. The ability of this strain to produce phytohormone indole-3-acetic acid from different carbon sources was studied. Indole-3-acetic acid synthesis by these transconjugants was 15-30 times as much in contrast to a wild-type strain with glucose as the sole carbon source. No difference was observed in other carbon or nitrogen sources. It is suggested that naphthalene dioxygenase is involved in the conversion of indole-3-pyruvic acid to indole-3-acetic acid. PMID- 11034293 TI - Protoplast preparation and regeneration from spores of the biocontrol fungus Pseudozyma flocculosa. AB - This paper describes a specific protocol for yielding and regenerating protoplasts from spores of the recently described powdery mildew biocontrol agent Pseudozyma flocculosa. With this protocol, a large quantity of protoplasts was obtained from beta-mercaptoethanol-pretreated spores in 3-day-old cultures of P. flocculosa grown in YMPD. Enzymatic digestion was optimal with 0.5% Novozym 234 and 5% Glucanex prepared in 0.6 M KCl in 0.1 M citrate buffer. All liberated protoplasts fluoresced in the presence of fluorescein diacetate indicating that viability was nearly 100%. The regeneration rate was equally outstanding reaching 75% when 0.8 M sucrose was used as osmotic stabilizer in the regeneration medium. This protocol will find useful applications in genetic studies of this poorly characterized and understood biocontrol agent. PMID- 11034291 TI - Detection and quantification of tetracyclines by whole cell biosensors. AB - Three different mini-Tn5 plasmids, containing a tetracycline-inducible promoter, Ptet and a regulatory gene, tetR, in operon fusions with a reporter gene system (lacZYA, luxCDABE or gfp), were constructed. These biosensor constructs responded to low levels of tetracyclines by producing beta-galactosidase, light or green fluorescent protein. They did so in a quantitative manner, thus enabling the quantification of tetracyclines in the immediate surroundings of the biosensor organism. All three constructs were transferred successfully to different gram negative bacteria by conjugation. An Escherichia coli strain containing the Ptet lac construct was used to determine oxytetracycline in milk as a demonstration of the application of these biosensors. PMID- 11034294 TI - Prevalence of HEp-2 cell-adherent Escherichia coli and characterisation of enteroaggregative E. coli and chain-like adherent E. coli isolated from children with and without diarrhoea, in Londrina, Brazil. AB - A total of 919 Escherichia coli isolates from 125 children with diarrhoea (cases) and 98 controls were assayed for adherence to HEp-2 cells. Localised adherence was found only in isolates from cases. Diffuse, aggregative (AA), chain-like adherence (CLA) and variants of the AA pattern were found in both cases and controls. The AA isolates were tested for gene sequences associated with enteroaggregative E. coli (EAEC). Only 25% of the isolates hybridised with the EAEC probe, and the aafA, astA and pet gene sequences were found in 7.9%, 44.7% and 7.9% of the isolates, respectively. The aggA gene was not found, although 7.9% were positive for aggC. The CLA isolates reacted with the EAEC probe (55.6%), and the aggC, astA and pet gene sequences were found in 66.7%, 33.3% and 11.1%, respectively. The aggR (55.6%), aspU (55.6%), shf (33.3%) and she (22.2%) genes were also found in CLA isolates. PMID- 11034295 TI - Novel arbekacin- and amikacin-modifying enzyme of methicillin-resistant Staphylococcus aureus. AB - An aminoglycoside-modifying enzyme in arbekacin-resistant methicillin-resistant Staphylococcus aureus (MRSA), exhibiting 4'''-N-acetylation, was examined. Although the MRSA strain with AAC(4''') had no AAC(6')-APH(2") activity, a DNA fragment of the AAC(6')-APH(2") gene was amplified by PCR and the purified N terminal 30-amino acid sequence of this AAC(4''') was identical to AAC(6') APH(2"). Direct DNA sequencing of this 'silent' AAC(6')-APH(2") gene revealed a single point mutation leading to a substitution of Gly for Asp80, through which the secondary structure is affected. A change in protein conformation could lead to a cleavage and a change of the enzymatic activity. We propose a new aminoglycoside-resistance mediated by AAC(4''') is caused by a mutation-modified AAC(6')-APH(2"). PMID- 11034296 TI - Production kinetics of acidophilin 801, a bacteriocin produced by Lactobacillus acidophilus IBB 801. AB - Lactobacillus acidophilus IBB 801 produces a small bacteriocin, designated acidophilin 801. Studying the relationship between growth and bacteriocin biosynthesis revealed primary metabolite kinetics of bacteriocin production with a peak activity at the end of the exponential growth phase followed by a decrease during the stationary phase. Both microbial growth and bacteriocin production was inhibited by lactic acid. Whereas volumetric bacteriocin production (activity units (AU) ml(-1)) was favoured under pH-controlled conditions, bacteriocin titres rapidly decreased because of strong adsorption of the bacteriocin molecules to the producing cells under less acidic conditions. PMID- 11034297 TI - Recombinant BCG approach for development of vaccines: cloning and expression of immunodominant antigens of M. tuberculosis. AB - In spite of major advances in our understanding of the biology and immunology of tuberculosis, the incidence of the disease has not reduced in most parts of the world. In an attempt to improve the protective efficacy of Mycobacterium bovis bacille Calmette-Guerin (BCG), we have developed a generic vector system, pSD5, for expression of genes at varying levels in mycobacteria. In this study, we have cloned and overexpressed three immunodominant secretory antigens of M. tuberculosis, 85A, 85B and 85C, belonging to the antigen 85 complex. All the genes were cloned under the control of a battery of mycobacterial promoters of varying strength. The expression was analysed in the fast-growing strain M. smegmatis and the slow-growing vaccine strain M. bovis BCG. The recombinant BCG constructs were able to express the antigens at high levels and the majority of the expressed antigens was secreted into the medium. These results show that by using this strategy the recombinant BCG approach can be successfully used for the development of candidate vaccines against infections associated with mycobacteria as well as other pathogens. PMID- 11034298 TI - Neither the A- nor B-repeat regions of the fibrinogen-binding protein of Streptococcus equi subsp. equi are essential for fibrinogen binding. AB - The major cell wall-associated protein (FgBP) of Streptococcus equi subsp. equi possesses two internal blocks of repeated sequence (A and B) and binds horse fibrinogen (Fg) avidly through residues located in the N-terminal half of the molecule. In the present study, we investigated the roles of the two repeats blocks in Fg binding through construction of recombinant FgBP proteins containing defined internal deletions of sequence. Ligand binding experiments clearly showed that neither repeat is essential for Fg binding. However, residues within the B repeats seem to play a major role in the aberrant mobility observed for FgBP following sodium dodecyl sulfate polyacrylamide gel electrophoresis. PMID- 11034299 TI - Interaction of Listeria monocytogenes with the intestinal epithelium. AB - Listeria monocytogenes is a food-borne pathogen that must cross the intestinal epithelial barrier to reach its target organs. We have investigated the importance of M cells in translocation using an experimental mouse model and a novel, recently described in vitro coculture system that mimics the follicle associated epithelium (FAE). Our data demonstrate that L. monocytogenes does not require, nor specifically use, M cells of the FAE to cross the gut. We also show that bacterial translocation is rapid and L. monocytogenes can attach very efficiently to exposed basal lamina of the small intestine indicating an important role for extracellular matrix proteins. PMID- 11034300 TI - Tools for discovery of inhibitors of the 1-deoxy-D-xylulose 5-phosphate (DXP) synthase and DXP reductoisomerase: an approach with enzymes from the pathogenic bacterium Pseudomonas aeruginosa. AB - Two Pseudomonas aeruginosa genes encoding the enzymes 1-deoxy-D-xylulose 5 phosphate (DXP) synthase and DXP reductoisomerase, both involved in the mevalonate-independent biosynthesis of isoprenoids, have been expressed as recombinant enzymes in Escherichia coli. The purified P. aeruginosa DXP reductoisomerase was inhibited by submicromolar concentrations of the antibiotics fosmidomycin and FR-900098 in a well established method. A novel and convenient spectrophotometric assay was developed to determine activity and inhibition of P. aeruginosa DXP synthase. Fluoropyruvate is described as a first inhibitor of DXP synthase. PMID- 11034301 TI - A Lactobacillus helveticus plasmid detects restriction fragment length polymorphism in different bacterial species. AB - A small cryptic Lactobacillus helveticus plasmid, pLBL4, was able to reveal restriction fragment length polymorphism in different bacterial species including Lactobacillus species, Bacillus species, and Escherichia coli when used as a DNA probe. The observed polymorphism was a result of the combined hybridization of several microsatellite sequences. The 6-bp sequence (TTGTTT) was repeated 12 times, seven of which were concentrated within the region between 1791 and 1997 bp of the plasmid sequence. The polymorphic patterns generated with pLBL4 differed from those obtained with M13 DNA in the larger number of bands observed. The results presented here open the possibility of using pLBL4 as a new broad spectrum polymorphic DNA probe for fingerprint analysis. PMID- 11034302 TI - Molecular characterisation of emergent multiresistant Salmonella enterica serotype [4,5,12:i:-] organisms causing human salmonellosis. AB - Salmonella multidrug-resistant clinical organisms identified as serotype [4,5,12:i:-] were typed using selected genetic procedures and compared with typhimurium organisms collected in the same Spanish region. Results showed a low genetic heterogeneity among [4,5,12:i:-] organisms, which generated identical ribotypes and similar but not identical XbaI PFGE, RAPD, and plasmid profiles. Multidrug resistance could be eliminated by curing and seems to be mediated by 140-kb (spvC+) and 120-kb (spvC-) non-self-transferable plasmids. The [4,5,12:i: ] organisms fall into a single genetic lineage, which emerged in 1997 and presents a different degree of genetic relationship with typhimurium lineages. PMID- 11034303 TI - Pigment-protein architecture in the light-harvesting antenna complexes of purple bacteria: does the crystal structure reflect the native pigment-protein arrangement? AB - Structural analysis of crystallized peripheral (LH2) and core antenna complexes (LH1) of purple bacteria has revealed circular aggregates of high rotational symmetry (C8, C9 and C16, respectively). Quantum-chemical calculations indicate that in particular the waterwheel-like arrangements of pigments should show characteristic structure-sensitive spectroscopic behavior in the near infrared absorption region. Laser-spectroscopic data obtained with non-crystallized, isolated LH2 of Rhodospirillum molischianum are in line with a highly symmetric (C8) circular aggregate, but deviations have been found for LH2 of Rhodobacter sphaeroides and Rhodopseudomonas acidophila. For both the latter, C-shaped incomplete circular aggregates (as seen only recently in electron micrographs of crystallized LH1-reaction center complexes) may be a suitable preliminary model. PMID- 11034304 TI - Characteristics of super alphaA-crystallin, a product of in vitro exon shuffling. AB - alphaA-Crystallin, a small heat shock protein with chaperone-like activity, forms dynamic multimeric complexes. Recently we described the spontaneous generation of a mutant protein (super alphaA-crystallin) by exon duplication arisen via exon shuffling confirming a classic hypothesis by Gilbert [Nature 271 (1978) 501]. Comparison of super alphaA-crystallin, which is viable in a mouse skeletal muscle cell line, with normal alphaA-crystallin shows that it has diminished thermostability, increased exposure of hydrophobic patches, a larger complex size and lost its chaperone activity. However, super alphaA-crystallin subunits exchange as readily between complexes as does normal alphaA-crystallin. These data indicate that chaperone-like activity may vanish independent of subunit hydrophobicity and exchangeability. PMID- 11034305 TI - Nuclear factor Y activates the human xanthine oxidoreductase gene promoter. AB - To study the regulation of the human xanthine oxidoreductase (XOR) gene, we cloned 1.9 kb of the promoter region. In reporter gene assays, a construct encompassing nucleotides between 142 to +42 conferred maximal basal activity of the XOR promoter in 293T cells, in comparison with shorter (-92 to +42) or longer (up to -1937 to +42) constructs. The promoter activity was low in NIH-3T3 cells. The most active construct contained a putative CCAAT motif at -119 to -123. Electrophoretic mobility shift assays showed that this sequence binds the ubiquitous nuclear factor Y (NF-Y). Mutation of the CCAAT motif (CTGAT) abolished the NF-Y binding and considerably reduced the promoter activity. Our data suggest an important functional role for NF-Y in the transcriptional activation of the human XOR gene. PMID- 11034306 TI - Ca2+-reversible inhibition of the mitochondrial megachannel by ubiquinone analogues. AB - Ubiquinone 0 and decylubiquinone have been reported to inhibit the mitochondrial permeability transition pore (PTP) [Fontaine, E., Ichas, F. and Bernardi, P. (1998) J. Biol. Chem. 273, 25734-257401, offering a new clue to its molecular composition. In patch-clamp experiments on rat liver mitochondria we have observed that these compounds also inhibit the previously described mitochondrial megachannel (MMC), confirming its identification as the PTP. Inhibition can be reversed by increasing [Ca2+], in analogy to the behavior observed with several other disparate PTP/MMC inhibitors. To rationalize the ability of Ca2+ to overcome inhibition by various quite different compounds we propose that it acts via the phospholipid bilayer. PMID- 11034307 TI - Differential gene expression in mesothelioma. AB - To investigate the molecular events controlling malignant transformation of human pleural cells, we compared constitutive gene expression of mesothelioma cells to that of pleural cells. Using cDNA microarray and high-density filter array, we assessed expression levels of > 6500 genes. Most of the highly expressed transcripts were common to both cell lines and included genes associated with stress response and DNA repair, outcomes consistent with the radio- and chemo resistance of mesothelioma. Interestingly, of the fewer than 300 genes that differed between cell lines, most functioned in (i) macromolecule stability, (ii) cell adhesion and recognition, (iii) cell migration (invasiveness), and (iv) extended cell division. Expression levels of several of these genes were confirmed by RT-PCR and could be useful as diagnostic markers of human mesothelioma. PMID- 11034308 TI - Activin A increases Pax4 gene expression in pancreatic beta cell lines. AB - Activin A, a member of the TGFbeta superfamily, has many physiological and developmental functions. In pancreatic beta cell cultures, activin promotes cell differentiation and insulin production. The author has found activin increases gene expression of the PAX4, one of the major transcription factors determining pancreatic beta cell differentiation. This effect was mediated, at least in part, by the type IB activin receptor (ALK4). Moreover, the activity of human insulin promoter-reporter system was controlled by PAX4 and its isoform PAX4 delta(G239 P251) in a unique fashion; positively by low concentrations, and negatively by high concentrations of these proteins. And the repression activities were different between these proteins. These findings confirm the importance of activin signal transduction in pancreatic beta cell development and function. PMID- 11034309 TI - Observation of DNA-polymer condensate formation in real time at a molecular level. AB - Dynamic real time assembly of toroidal and rod-like DNA condensates has been visualised using atomic force microscopy. Imaging has been conducted in an aqueous environment allowing the visualisation of hydrated, pegylated-polymer DNA condensates undergoing dynamic structural movement and conformational change. A major hurdle in the field of gene delivery is cellular transfection and the subsequent transfer of condensed genetic material to the cell nucleus. An increased understanding of the process of DNA condensation will aid the development and optimisation of gene delivery vectors. PMID- 11034310 TI - Expression of Mona (monocytic adapter) in myeloid progenitor cells results in increased and prolonged MAP kinase activation upon macrophage colony-stimulating factor stimulation. AB - Mona is an SH3 and SH2 domain-containing adapter molecule that is induced during monocytic differentiation. Here we have first shown that M-CSFR is the major Mona partner in M-CSF signaling, the interaction being mediated through tyrosine 697 of the receptor. Next we asked whether Mona expression would alter the Ras/MAP kinase pathway since Mona is a likely competitor of Grb2 for binding to M-CSFR. We found that M-CSF induced late and massive phosphorylation of ERK molecules in Mona-expressing myeloid cells compared to non-expressing cells. These results suggest that Mona expression might modify M-CSF signaling during monocytic differentiation. PMID- 11034311 TI - A novel type of FKBP in the secretory pathway of Neurospora crassa. AB - FKBPs define a subfamily of peptidyl-prolyl cis/trans isomerases (PPlases). PPlases are known to play roles in cellular protein folding, protein interactions and signal transduction. Here we describe NcFKBP22 from Neurospora crassa, a novel type of FKBP. NcFKBP22 is synthesized as a precursor protein with a cleavable signal sequence. In addition to a typical FKBP domain in the amino terminal part mature NcFKBP22 contains a novel second domain which is unique amongst all known FKBPs. The amino acid composition of this carboxyterminal domain is highly biased. Secondary structure predictions suggest that this domain may form an amphipathic alpha-helix. The carboxy-terminus of NcFKBP22 is -HNEL, a potential endoplasmic reticulum (ER) retention signal, suggesting that NcFKBP22 is a resident protein of the ER. PMID- 11034312 TI - The role of chromophore in the lipid-protein interactions in bacteriorhodopsin phosphatidylcholine vesicles. AB - By fluorescence and phase properties of a 1-acyl-2-[8-(2-anthroyl)-octanoyl]-sn glycero-3-phosphocholine probe, the influence of the chromophore on the phase transition of bacteriorhodopsin-lipid vesicles was investigated. It was observed that removal of the chromophore led to the down-shifting of the phase transition temperatures. The temperatures corresponding to the beginning and ending of the gel-liquid phase transition were also influenced. This demonstrated that the liquid phase is reached more easily when the chromophore is bleached. The results indicate that removal of the chromophore alters the protein-lipid interactions. It is suggested that this alteration might be related to the change in the lipid molecular packing. PMID- 11034313 TI - 31P-CP-MAS NMR studies on TPP+ bound to the ion-coupled multidrug transport protein EmrE. AB - The binding of tetraphenylphosphonium (TPP+) to EmrE, a membrane-bound, 110 residue Escherichia coli multidrug transport protein, has been observed by 31P cross-polarisation-magic-angle spinning nuclear magnetic resonance spectroscopy (CP-MAS NMR). EmrE has been reconstituted into dimyristoyl phosphatidylcholine bilayers. CP-MAS could selectively distinguish binding of TPP+ to EmrE in the fluid membrane. A population of bound ligand appears shifted 4 ppm to lower frequency compared to free ligand in solution, which suggests a rather direct and specific type of interaction of the ligand with the protein. This is also supported by the observed restricted motion of the bound ligand. The observation of another weakly bound substrate population arises from ligand binding to negatively charged residues in the protein loop regions. PMID- 11034314 TI - IL-10 attenuates IFN-alpha-activated STAT1 in the liver: involvement of SOCS2 and SOCS3. AB - Interleukin-10 (IL-10) has been used in the treatment of viral hepatitis in interferon-alpha (IFN-alpha) non-responders while patients who have high levels of IL-10 are poorly responsive to IFN-alpha. The mechanism underlying such controversial functions of IL-10 remains unknown. Here we demonstrated that injection of IL-10 into mice attenuated IFN-alpha-induced signal transducer and activator transcription factor (STAT)1 tyrosine phosphorylation in the liver. Reverse transcriptase-polymerase chain reaction assay demonstrated that mouse liver expressed high levels of IL-10 receptor 2 (IL-10R2) but low levels of IL 10R1. Injection of IL-10 into mice activated STAT3 but not STAT1 tyrosine phosphorylation and induced suppressor of cytokine signal 2 (SOCS2), SOCS3, and cytokine-inducible SH2 protein (CIS) mRNA expression in the liver. Furthermore, overexpression of SOCS2 or SOCS3 inhibited IFN-alpha-induced reporter activity in hepatic cells. These findings suggest that IL-10 inhibits IFN-alpha-activated STAT1 in the liver, at least in part, by inducing SOCS2, SOCS3, and CIS expression, which may be responsible for the resistance of IFN-alpha therapy in patients who have high levels of IL-10 and recommends that IL-10 treatment for viral hepatitis should be cautious. PMID- 11034315 TI - M-type KCNQ2-KCNQ3 potassium channels are modulated by the KCNE2 subunit. AB - KCNQ2 and KCNQ3 subunits belong to the six transmembrane domain K+ channel family and loss of function mutations are associated with benign familial neonatal convulsions. KCNE2 (MirP1) is a single transmembrane domain subunit first described to be a modulator of the HERG potassium channel in the heart. Here, we show that KCNE2 is present in brain, in areas which also express KCNQ2 and KCNQ3 channels. We demonstrate that KCNE2 associates with KCNQ2 and/or KCNQ3 subunits. In transiently transfected COS cells, KCNE2 expression produces an acceleration of deactivation kinetics of KCNQ2 and of the KCNQ2-KCNQ3 complex. Effects of two previously identified arrhythmogenic mutations of KCNE2 have also been analyzed. PMID- 11034316 TI - Membrane-type 6 matrix metalloproteinase (MT6-MMP, MMP-25) is the second glycosyl phosphatidyl inositol (GPI)-anchored MMP. AB - A recently identified membrane-type 6 matrix metalloproteinase (MT6-MMP) has a hydrophobic stretch of 24 amino acids at the C-terminus. This hydrophobicity pattern is similar to glycosyl-phosphatidyl inositol (GPI)-anchored MMP, MT4-MMP, and other GPI-anchored proteins. Thus, we tested the possibility that MT6-MMP was also a GPI-anchored proteinase. Our results showed that MT6-MMP as well as MT4 MMP were labeled with [3H]ethanolamine indicating the presence of a GPI unit with incorporated label. In addition, phosphatidyl inositol-specific phospholipase C treatment released MT6-MMP from the surface of transfected cells. These results strongly indicate that MT6-MMP is a GPI-anchored protein. Since two members of MT MMPs are now assigned as GPI-anchored proteinase, MT-MMPs can be subgrouped into GPI type and transmembrane type. PMID- 11034317 TI - LEAP-1, a novel highly disulfide-bonded human peptide, exhibits antimicrobial activity. AB - We report the isolation and characterization of a novel human peptide with antimicrobial activity, termed LEAP-1 (liver-expressed antimicrobial peptide). Using a mass spectrometric assay detecting cysteine-rich peptides, a 25-residue peptide containing four disulfide bonds was identified in human blood ultrafiltrate. LEAP-1 expression was predominantly detected in the liver, and, to a much lower extent, in the heart. In radial diffusion assays, Gram-positive Bacillus megaterium, Bacillus subtilis, Micrococcus luteus, Staphylococcus carnosus, and Gram-negative Neisseria cinerea as well as the yeast Saccharomyces cerevisiae dose-dependently exhibited sensitivity upon treatment with synthetic LEAP-1. The discovery of LEAP-1 extends the known families of mammalian peptides with antimicrobial activity by its novel disulfide motif and distinct expression pattern. PMID- 11034318 TI - Extremely efficient cleavage of eIF4G by picornaviral proteinases L and 2A in vitro. AB - Certain picornaviruses encode proteinases which cleave the translation initiation factor eIF4G, a member of the eIF4F complex which recruits mRNA to the 40S ribosomal subunit during initiation of protein synthesis in eukaryotes. We have compared the efficiency of eIF4G cleavage in rabbit reticulocyte lysates during translation of mRNAs encoding the foot-and-mouth disease virus leader proteinase (Lpro) or the human rhinovirus 2Apro. Under standard translation conditions, Lpro cleaved 50% of eIF4G within 4 min after initiation of protein synthesis, whereas 2Apro required 15 min. At these times, the molar ratios of proteinase to eIF4G were 1:130 for Lpro and 1:12 for 2Apro, indicating a much more efficient in vitro cleavage than previously observed. The molar ratios are similar to those observed during viral infection in vivo. PMID- 11034320 TI - Iron(II) induces changes in the conformation of mammalian mitochondrial DNA resulting in a reduction of its transcriptional rate. AB - Living isolated mitochondria incubated with iron(II) show a major alteration in mitochondrial DNA (mtDNA) conformational forms as assessed by Southern blot analysis of undigested mtDNA. In the presence of iron(II), form I is transformed into form III in a dose-dependent manner. This alteration in mtDNA conformation shows a strong correlation with a decrease in the mtDNA transcription rate (r=0.965, P < 0.002), suggesting that iron(II) load results in double-strand breaks and unwinding of mtDNA, which, in turn, is unable to maintain its normal transcriptional rate. PMID- 11034319 TI - A delta opioid receptor lacking the third cytoplasmic loop is generated by atypical mRNA processing in human malignomas. AB - delta Opioid receptors were identified in human melanomas by RT-PCR and radioligand binding. In all tumors an additional PCR amplificate was detected in which 144 bp within the third exon were deleted. This fragment corresponded to the third cytoplasmic domain of the receptor protein. The short variant resulted from atypical mRNA processing. There were no common splice recognition sequences around the deleted fragment; instead its excision resembled the removal of a transposon. The deletion was not detected in normal human melanocytes nor in human or rat brain. However, it was present in a human neuroblastoma cell line (SH-SY5Y). Thus, it appears that the occurrence of the short delta opioid receptor is correlated to malignancy. PMID- 11034321 TI - Cleavage of lamin-like proteins in in vivo and in vitro apoptosis of tobacco protoplasts induced by heat shock. AB - Apoptosis in heat shock-treated tobacco protoplasts was evidenced by DNA fragmentation, flow cytometric analysis and activation of caspase 3-like protease. Furthermore, an in vitro apoptosis system was established which reproduced the apoptotic events. Western blotting analysis using an antibody against lamin A and C showed that in both in vivo and in vitro systems lamin-like proteins were cleaved into a 35-kDa fragment, and that lamin-like protein degradation precedes DNA fragmentation. Moreover, we found a 22.8-fold increase in caspase 6-like activity in cytosol of heat-treated protoplasts as compared with the control. PMID- 11034322 TI - Evidence showing that the two-chain form of vitronectin is produced in the liver by a selective furin cleavage. AB - The adhesive protein vitronectin (75 kDa) occurs in human blood fluid in a one chain (Vn75) or a two-chain form (Vn65-10), and is produced by a specific cleavage (at Arg379-Ala380), by a proteinase not identified hitherto. These two forms were shown to be functionally different and therefore, this cleavage may have a regulatory significance in vivo. Here, we report the use of a tailored one chain recombinant Vn, a specific protein kinase A phosphorylation at Ser378, and sequence analysis to show: (1) that none of the proteinases originating from blood, previously thought to be the endogenous proteinase (plasmin, thrombin, tPA, and uPA), is indeed the in vivo convertase; and (2) that furin, a serine endoproteinase residing in the secretory pathway of hepatocytes, where Vn is synthesized, specifically cleaves Vn at the endogenous cleavage site. Consequently, we propose that the Vn75 to Vn65-10 conversion takes place in the liver (not in blood) and is carried out by furin. PMID- 11034323 TI - Propensities for the formation of individual disulfide bonds in hen lysozyme and in the size and stability of disulfide-associated submolecular structures. AB - Hen lysozyme single-disulfide variants were constructed to characterize the structures associated with the formation of individual native disulfide bonds. Circular dichroism spectra and the effective concentration of protein thiol groups showed that the propensity for structure formation was relatively high for Cys-6-Cys-127 and Cys-30-Cys-115 disulfides. The urea concentration dependence of individual effective concentrations showed that the apparent sizes of the structures were 14-50% of the whole molecule. The intrinsic stability of each submolecular structure in a reduced form of protein, obtained by subtracting the entropic contribution of cross-linking, was highest for Cys-64-Cys-80 and lowest for Cys-76-Cys-94 disulfide bonds. PMID- 11034324 TI - Effect of dimethyl adipimidate on K+ transport and shape change in red blood cells from sickle cell patients. AB - Dimethyl adipimidate (DMA) reduces K+ loss from, and dehydration of, red cells containing haemoglobin S (HbS cells). Three membrane transporters may contribute to these processes: the deoxygenation-induced cation-selective channel (Psickle), the Ca2+-activated K+ channel (or Gardos channel) and the K+-CI- cotransporter (KCC). We show that DMA inhibited all three pathways in deoxygenated HbS cells. The Gardos channel could be activated following Ca2+ loading. Considerable KCC activity was present in oxygenated HbS cells, showing a selective action of DMA on the transporter in deoxygenated cells. Inhibition of sickling correlated strongly with that of Psickle and moderately with that of KCC activity. We conclude that DMA does not inhibit the K+ pathways directly, but acts mainly by preventing HbS polymerisation and sickling. These findings are relevant to the development of novel chemotherapeutic agents for amelioration of sickle cell disease. PMID- 11034325 TI - Regulation of exocytosis in chromaffin cells by phosducin-like protein, a protein interacting with G protein betagamma subunits. AB - Phosducin and related proteins have been identified as ubiquitous regulators of signalling mediated by betagamma subunits of trimeric G proteins. To explore a role for phosducin in regulated exocytosis, we have examined the distribution and putative function of phosducin-like protein (PhLP) in adrenal medullary chromaffin cells. The full-length cDNA encoding the short splice variant of PhLP (PhLPs) was cloned from cultured chromaffin cells. Native PhLPs was found associated with plasma membranes and detected in the subplasmalemmal area of resting chromaffin cells by confocal immunofluorescence analysis. Stimulation with secretagogues triggered a massive redistribution of PhLPs into the cytoplasm. When microinjected into individual chromaffin cells, recombinant PhLPs inhibited catecholamine secretion evoked by a depolarizing concentration of K+ without affecting calcium mobilization. Thus, PhLPs may participate directly in the regulation of calcium-evoked exocytosis. PMID- 11034326 TI - Tenascin-C induced stimulation of chondrogenesis is dependent on the presence of the C-terminal fibrinogen-like globular domain. AB - The relationship between structure of tenascin-C (Tn-C), a multi-domain extracellular matrix protein, and its stimulation of chondrogenesis was examined using recombinant Tn-C isoforms (full length or with specific domains deleted) as substrata for undifferentiated chicken mesenchymal cells. Of the Tn-C variants tested, only Tn-C lacking the fibrinogen-like domain or Tn-C comprised solely of fibrinogen-like domains failed to stimulate chondrogenesis. The ability of variants to stimulate chondrogenesis was not dependent on their ability to support adhesion or stimulate proliferation. These results demonstrate that the fibrinogen-like domain of Tn-C is necessary but not sufficient for induction of chondrogenesis. PMID- 11034327 TI - Aspirin induces apoptosis through mitochondrial cytochrome c release. AB - Aspirin and other non-steroidal anti-inflammatory drugs induce apoptosis in many cell types. Although the involvement of caspases has been demonstrated, the mechanism leading to caspase activation remains unknown. We have studied the role of the mitochondrial pathway in aspirin-induced apoptosis. The apoptotic effect of aspirin was analyzed in different cell lines (Jurkat, MOLT-4, Raji and HL-60) showing induction of mitochondrial cytochrome c release and caspases 9, 3 and 8 processing. Furthermore, early aspirin-induced cytochrome c release was not affected by the caspase inhibitor Z-VAD x fmk and preceded loss of mitochondrial membrane potential. Therefore, aspirin-induced apoptosis involves caspase activation through cytochrome c release. PMID- 11034328 TI - Involvement of apoptosis-inducing factor during dolichyl monophosphate-induced apoptosis in U937 cells. AB - Dolichyl monophosphate (Dol-P) has been found to induce apoptosis in human leukemia U937 cells. During this apoptotic execution, the increase of plasma membrane fluidity (5-20 min), caspase-3-like protease activation (2-4 h), chromatin condensation and DNA ladder formation (3-4 h) were observed successively. Here, we report that reduction in mitochondrial transmembrane potential and translocation of apoptosis-inducing factor (AIF) are early events (1-3 h) in the apoptotic process induced by Dol-P in U937 cells. The AIF was concentrated around nuclei and partly translocated to the nuclei, which was confirmed by immunocytochemistry using specific anti-AIF antibody. Both caspase-8 and caspase-3 inhibitors blocked only DNA fragmentation but not mitochondrial processes, AIF migration and chromatin condensation. These results indicate that mitochondrial changes are an early step in the apoptosis induced by Dol-P and AIF is one of the important factors which induce chromatin condensation in nuclei. PMID- 11034329 TI - N-terminal and core-domain random mutations in human topoisomerase II alpha conferring bisdioxopiperazine resistance. AB - Random mutagenesis of human topoisomerase II alpha cDNA followed by functional expression in yeast cells lacking endogenous topoisomerase II activity in the presence of ICRF-187, identified five functional mutations conferring cellular bisdioxopiperazine resistance. The mutations L169F, G551S, P592L, D645N, and T996L confer > 37, 37, 18, 14, and 19 fold resistance towards ICRF-187 in a 24 h clonogenic assay, respectively. Purified recombinant L169F protein is highly resistant towards catalytic inhibition by ICRF-187 in vitro while G551S, D645N, and T996L proteins are not. This demonstrates that cellular bisdioxopiperazine resistance can result from at least two classes of mutations in topoisomerase II; one class renders the protein non-responsive to bisdioxopiperazine compounds, while an other class does not appear to affect the catalytic sensitivity towards these drugs. In addition, our results indicate that different protein domains are involved in mediating the effect of bisdioxopiperazine compounds. PMID- 11034330 TI - In vitro nuclear reconstitution could be induced in a plant cell-free system. AB - A cell-free system derived from carrot cell cytosol extract has been developed for reassembling nuclear structure around the added demembranated sperm chromatin of Xenopus. Morphological evidence suggests that reassembled nuclei display the typical characteristics of normal eukaryotic nuclei, such as double-layered nuclear membrane and nuclear pores. Micrococcal nuclease treatment indicates that remodeling of the demembranated sperm chromatin has occurred and the structure of nucleosome is formed during nuclear reconstitution. These data indicate that the nuclear reconstitution can be induced in cell-free systems from plants, and the self-assembly of the nucleus is ubiquitous in both animal and plant cells. PMID- 11034331 TI - Iron-coproporphyrin III is a natural cofactor in bacterioferritin from the anaerobic bacterium Desulfovibrio desulfuricans. AB - A bacterioferritin was recently isolated from the anaerobic sulphate-reducing bacterium Desulfivibrio desulfuricans ATCC 27774 [Romao et al. (2000) Biochemistry 39, 6841-6849]. Although its properties are in general similar to those of the other bacterioferritins, it contains a haem quite distinct from the haem B, found in bacterioferritins from aerobic organisms. Using visible and NMR spectroscopies, as well as mass spectrometry analysis, the haem is now unambiguously identified as iron-coproporphyrin III, the first example of such a prosthetic group in a biological system. This unexpected finding is discussed in the framework of haem biosynthetic pathways in anaerobes and particularly in sulphate-reducing bacteria. PMID- 11034332 TI - Binding to 14-3-3 proteins is not sufficient to inhibit nitrate reductase in spinach leaves. AB - To assess the role of 14-3-3 proteins in the magnesium-dependent inhibition of nitrate reductase (NR) we tested the effect of magnesium on NR binding to 14-3-3s by coimmunoprecipitation and gel filtration. The stability of the 14-3-3 complex of NR was, unlike its activity, unaffected by magnesium. We therefore conclude that binding to 14-3-3s per se does not inhibit NR. Magnesium inhibited 14-3-3 bound NR much more strongly than 14-3-3-free NR. 14-3-3s possibly reinforce NR inhibition by magnesium. PMID- 11034333 TI - Inteins invading mycobacterial RecA proteins. AB - Five new inteins were discovered in a survey of 39 mycobacterial strains that was undertaken to clarify the role of RecA inteins in mycobacteria. They are all inserted at the RecA-b site of the recA gene of Mycobacterium chitae, 4. fallax, M. gastri, M. shimodei and M. thermoresistibile and belong to the MleRecA allelic family. Sequence analysis showed that although only M. tuberculosis harbours an intein at the RecA-a site the sequence of the RecA-b site is well conserved between species. Furthermore, the presence of inteins does not correlate with specific characteristics of the species such as pathogenicity or growth rate. PMID- 11034334 TI - Assembly of the CD8alpha/p56(lck) protein complex in stably expressing rat epithelial cells. AB - We have previously characterized the biogenesis of the human CD8alpha protein expressed in rat epithelial cells. We now describe the biosynthesis, post translational maturation and hetero-oligomeric assembly of the human CD8alpha/p56(lck) protein complex in stable transfectants obtained from the same cell line. There were no differences in the myristilation of p56(lck), or in the dimerization, O-glycosylation and transport to the plasma membrane of CD8alpha, between cells expressing either one or both proteins. In the doubly expressing cells, dimeric forms of CD8alpha established hetero-oligomeric complexes with p56(lck), as revealed by co-immunoprecipitation assays performed with anti CD8alpha antibody. Moreover, p56(lck) bound in these hetero-oligomeric complexes was endowed with auto- and hetero-phosphorylating activity. The present study shows that: (1) the newly synthesized p56(lck) binds rapidly to CD8alpha and most of the p56(lck) is bound to CD8alpha at steady state; (2) CD8alpha/p56(lck) protein complexes are formed at internal membranes as well as at the plasma membrane; and (3) about 50% of complexed p56(lck) reaches the cell surface. PMID- 11034336 TI - Both native conformers of rabbit muscle adenylate kinase are active. AB - There are two forms of rabbit muscle adenylate kinase (AK) with different 8 anilino-1-naphthalenesulfonic acid (ANS) binding properties in equilibrium solution. One form (about 70%, denoted N1) binds rapidly with ANS, whereas the other (about 30%, denoted N2) does not. Furthermore, native forms of AK should adopt different conformations for binding with substrates and products, which should be pre-existing for performing its catalytic function. The present experiments demonstrate both forms of AK distinguished by ANS probe are active. The activity of N2 is about 0.8 fold higher than N1 and shows higher susceptibility to proteolysis by trypsin. This means that the native state of AK might be an ensemble of kinetically attainable conformers and the energy landscapes of AK folding should be rugged with more than one local minimum. PMID- 11034335 TI - Efficient phage display of polypeptides fused to the carboxy-terminus of the M13 gene-3 minor coat protein. AB - We report that, contrary to common belief, polypeptides fused to the carboxy terminus of the M13 gene-3 minor coat protein are functionally displayed on the phage surface. In a phagemid display system, carboxy-terminal fusion through optimized linker sequences resulted in display levels comparable to those achieved with conventional amino-terminal fusions. These findings are of considerable importance to phage display technology because they enable investigations not suited to amino-terminal display, including the study of protein-protein interactions requiring free carboxy-termini, functional cDNA cloning efforts, and the display of intracellular proteins. PMID- 11034337 TI - Colocalization of leukotriene C synthase and microsomal glutathione S-transferase elucidated by indirect immunofluorescence analysis. AB - We have previously shown that the two membrane bound enzymes leukotriene C synthase and microsomal glutathione S-transferase interact in vitro and in vivo. Rat basophilic leukemia cells and murine mastocytoma cells, two well-known sources of leukotriene C synthase, both expressed microsomal glutathione S transferase as determined by Western blot analyses. Several human tissues were found to contain both leukotriene C synthase and microsomal glutathione S transferase mRNA. These data suggest that the interaction may be physiologically important. To study this further, expression vectors encoding the two enzymes were cotransfected into mammalian cells and the subcellular localization of the enzymes was determined by indirect immunofluorescence using confocal laser scanning microscopy. The results showed that leukotriene C synthase and microsomal glutathione S-transferase were both localized on the nuclear envelope and adjacent parts of the endoplasmic reticulum. Image overlay demonstrated virtually identical localization. We also observed that coexpression substantially reduced the catalytic activity of each enzyme suggesting that a mechanism involving protein-protein interaction may contribute to the regulation of LTC4 production. PMID- 11034338 TI - Bi-substrate analogue ligands for affinity chromatography of protein kinases. AB - Novel affinity ligands, consisting of ATP-resembling part coupled with specificity determining peptide fragment, were proposed for purification of protein kinases. Following this approach affinity sorbents based on two closely similar ligands AdoC-Aoc-Arg4-Lys and AdoC-Aoc-Arg4-NH(CH2)6NH2, where AdoC stands for adenosine-5'-carboxylic acid and Aoc for amino-octanoic acid, were synthesized and tested for purification of recombinant protein kinase A catalytic subunit directly from crude cell extract. Elution of the enzyme with MgATP as well as L-arginine yielded homogeneous protein kinase A preparation in a single purification step. Also protein kinase A from pig heart homogenate was selectively isolated using MgATP as eluting agent. Protein kinase with acidic specificity determinant (CK2) as well as other proteins possessing nucleotide binding site (L-type pyruvate kinase) or sites for wide variety of different ligands (bovine serum albumin) did not bind to the column, pointing to high selectivity of the bi-functional binding mode of the affinity ligand. PMID- 11034339 TI - C2PA, a new protein expressed during mouse spermatogenesis. AB - C2PA is a novel protein that contains a C2 membrane binding domain, a PDZ protein/protein interaction domain, and an ATP/GTP binding domain. C2PA is expressed during embryogenesis from 8.5 days post-coitum (dpc) until birth. After birth, C2PA expression is mainly observed in the post-natal and adult testis. During spermatogenesis, C2PA transcripts are specifically observed in the spermatocytes, whereas spermatogonia and spermatids are negative. Taken together, these results suggest that C2PA might be involved in cell signaling pathways occurring during spermatogenesis. PMID- 11034340 TI - A chloroplastic RNA-binding protein is a new member of the PPR family. AB - P67, a new protein binding to a specific RNA probe, was purified from radish seedlings [Echeverria, M. and Lahmy, S. (1995) Nucleic Acids Res. 23, 4963-4970]. Amino acid sequence information obtained from P67 microsequencing allowed the isolation of genes encoding P67 in radish and Airabidopsis thaliana. Immunolocalisation experiments in transfected protoplasts demonstrated that this protein is addressed to the chloroplast. The RNA-binding activity of recombinant P67 was found to be similar to that of the native protein. A significant similarity with the maize protein CRP1 [Fisk, D.G., Walker, M.B. and Barkan, A. (1999) EMBO J. 18, 2621-2630] suggests that P67 belongs to the PPR family and could be involved in chloroplast RNA processing. PMID- 11034341 TI - Cells overexpressing fructose-2,6-bisphosphatase showed enhanced pentose phosphate pathway flux and resistance to oxidative stress. AB - Changes in the content of fructose-2,6-bisphosphate, a modulator of glycolytic flux, also affect other metabolic fluxes such as the non-oxidative pentose phosphate pathway. Since this is the main source of precursors for biosynthesis in proliferating cells, PFK-2/FBPase-2 has been proposed as a potential target for neoplastic treatments. Here we provide evidence that cells with a low content of fructose-2,6-bisphosphate have a lower energy status than controls, but they are also less sensitive to oxidative stress. This feature is related to the activation of the oxidative branch of the pentose phosphate pathway and the increased production of NADPH. PMID- 11034342 TI - The bulk of UCP3 expressed in yeast cells is incompetent for a nucleotide regulated H+ transport. AB - The impact of uncoupling protein (UCP) 1, UCP3 and UCP3s expressed in yeast on oxidative phosphorylation, membrane potential and H+ transport is determined. Intracellular ATP synthesis is inhibited by UCP3, much more than by UCP1, while similar levels of UCP3 and UCP1 exist in the mitochondrial fractions. Measurements of membrane potential and H+ efflux in isolated mitochondria show that, different from UCP1, with UCP3 and UCP3s there is a priori a preponderant uncoupling not inhibited by GDP. The results are interpreted to show that UCP3 and UCP3s in yeast mitochondria are in a deranged state causing uncontrolled uncoupling, which does not represent their physiological function. PMID- 11034343 TI - A peroxidase homologue and novel plastocyanin located by proteomics to the Arabidopsis chloroplast thylakoid lumen. AB - A study by two-dimensional electrophoresis showed that the soluble, lumenal fraction of Arabidopsis thaliana thylakoids can be resolved into 300 protein spots. After subtraction of low-intensity spots and accounting for low-level stromal contamination, the number of more abundant, lumenal proteins was estimated to be between 30 and 60. Two of these proteins have been identified: a novel plastocyanin that also was the predominant component of the total plastocyanin pool, and a putative ascorbate peroxidase. Import studies showed that these proteins are routed to the thylakoid lumen by the Sec- and delta pH dependent translocation pathways, respectively. In addition, novel isoforms of PsbO and PsbQ were identified. PMID- 11034344 TI - Distinct isoforms of ADPglucose pyrophosphatase and ADPglucose pyrophosphorylase occur in the suspension-cultured cells of sycamore (Acer pseudoplatanus L.). AB - The intracellular localizations of ADPglucose pyrophosphatase (AGPPase) and ADPglucose pyrophosphorylase (AGPase) have been studied using protoplasts prepared from suspension-cultured cells of sycamore (Acer pseudoplatanus L.). Subcellular fractionation studies revealed that all the AGPPase present in the protoplasts is associated with amyloplasts, whereas more than 60% of AGPase is in the extraplastidial compartment. Immunoblots of amyloplast- and extraplastid enriched extracts further confirmed that AGPase is located mainly outside the amyloplast. Experiments carried out to identify possible different isoforms of AGPPase in the amyloplast revealed the presence of soluble and starch granule bound isoforms. We thus propose that ADPglucose levels linked to starch biosynthesis in sycamore cells are controlled by enzymatic reactions catalyzing the synthesis and breakdown of ADPglucose, which take place both inside and outside the amyloplast. PMID- 11034345 TI - Ulip6, a novel unc-33 and dihydropyrimidinase related protein highly expressed in developing rat brain. AB - Here, we report the identification of Ulip6, a novel unc-33 and dihydropyrimidinase related protein that belongs to the Ulip/CRMP protein family. Ulip6 was found in a yeast two-hybrid screen using the neuronal glycine transporter GlyT2 as bait. The rat and human Ulip6 sequences are highly homologous and most closely related to the liver enzyme dihydropyrimidinase (Ulip5). Northern and Western analysis of rat tissues revealed that the distribution of the Ulip6 mRNA and protein resembles those of brain-type Ulip proteins. Like Ulip1-4, Ulip6 is highly expressed in embryonic and early postnatal brain and spinal cord. These findings are consistent with Ulip6 having a function in neuronal differentiation and/or axon growth. PMID- 11034346 TI - The yeast exchange assay, a new complementary method to screen for Dbl-like protein specificity: identification of a novel RhoA exchange factor. AB - The target Rho GTPases of many guanine nucleotide exchange factors (GEFs) of the Dbl family remain to be identified. Here we report a new method: the yeast exchange assay (YEA), a rapid qualitative test to perform a wide range screen for GEF specificity. In this assay based on the two-hybrid system, a wild type GTPase binds to its effector only after activation by a specific GEF. We validated the YEA by activating GTPases by previously reported GEFs. We further established that a novel GEF, GEF337, activates RhoA in the YEA. GEF337 promoted nucleotide exchange on RhoA in vitro and promoted F-actin stress fiber assembly in fibroblasts, characteristic of RhoA activation. PMID- 11034347 TI - An unexpectedly large working stroke from chymotryptic fragments of myosin II. AB - Recent structural evidence indicates that the light chain domain of the myosin head (LCD) bends on the motor domain (MD) to move actin. Structural models usually assume that the actin-MD interface remains static and the possibility that part of the myosin working stroke might be produced by rotation about the acto-myosin interface has been neglected. We have used an optical trap to measure the movement produced by proteolytically shortened single rabbit skeletal muscle myosin heads (S-1(A1) and S-1(A2)). The working stroke produced by these shortened heads was more than that which the MD-LCD bend mechanism predicts from the full-length (papain) S-1's working stroke obtained under similar conditions. This result indicates that part of the working stroke may be caused by motor action at the actin-MD interface. PMID- 11034348 TI - Myosin light chain kinase binding to actin filaments. AB - Smooth muscle myosin light chain kinase (MLCK) plays important roles in contractile-motile processes of a variety of cells. Three DFRxxL motifs at the kinase N-terminus (residues 2-63) are critical for high-affinity binding to actin containing filaments [Smith et al. (1999) J. Biol. Chem. 274, 29433-29438]. A GST fusion protein containing residues 1-75 of MLCK (GST75-MLCK) bound maximally to both smooth muscle myofilaments and F-actin at 0.28 and 0.31 mol GST75-MLCK/mol actin with respective K(D) values of 0.1 microM and 0.8 microM. High-affinity binding of MLCK to actin-containing filaments may be due to each DFRxxL motif binding to one actin monomer in filaments. PMID- 11034349 TI - The phytotoxic lichen metabolite, usnic acid, is a potent inhibitor of plant p hydroxyphenylpyruvate dioxygenase. AB - The lichen secondary metabolite usnic acid exists as a (-) and a (+) enantiomer, indicating a alpha or beta projection of the methyl group at position 9b, respectively. (-)-Usnic caused a dose-dependent bleaching of the cotyledonary tissues associated with a decrease of both chlorophylls and carotenoids in treated plants whereas no bleaching was observed with the (+) enantiomer. (-) Usnic acid inhibited protophorphyrinogen oxidase activity (I50 = 3 microM), but did not lead to protoporphyrin IX accumulation. Bleaching appears to be caused by irreversible inhibition of the enzyme 4-hydroxyphenylpyruvate dioxygenase by (-) usnic acid (apparent IC50 = 50 nM). PMID- 11034350 TI - The influence of glycerol and chloroplast lipids on the spectral shifts of pigments associated with NADPH: protochlorophyllide oxidoreductase from Avena sativa L. AB - Dark-grown angiosperm seedlings lack chlorophylls, but accumulate protochlorophyllide a complexed with the light-dependent enzyme NADPH:protochlorophyllide oxidoreductase. Previous investigators correlated spectral heterogeneity of in vivo protochlorophyllide forms and a shift of chlorophyllide forms from 680 to 672 nm (Shibata shift) occurring after irradiation, with intact membrane structures which are destroyed by solubilization. We demonstrate here that the various protochlorophyllide forms and the Shibata shift which disappear upon solubilization are restored if the reconstituted complex is treated with plastid lipids and 80% (w/v) glycerol. We hypothesize that the lipids can form a cubic phase and that this is the precondition in vitro and in vivo for the observed spectral properties before and after irradiation. PMID- 11034351 TI - Evolutionarily conserved Bok proteins in the Bcl-2 family. PMID- 11034352 TI - Lipopolysaccharide induces physical proximity between CD14 and toll-like receptor 4 (TLR4) prior to nuclear translocation of NF-kappa B. AB - CD14, a GPI-linked protein, plays a pivotal role in LPS-mediated signaling by potentiating leukocyte adherence, activation, and cytokine production. Recent studies have identified the Toll-like receptor 4 (TLR4) as a membrane cofactor in LPS-mediated transmembrane signaling in cytokine induction, although the mechanism responsible for this cooperation is unknown. Using fluorescence resonance energy transfer (RET) techniques, we demonstrate that LPS triggers a physical association between CD14 and TLR4. Because LPS stimulation upregulates CD14 and TLR4 expression, it was necessary to control for the possibility that these newly expressed molecules were associated with one another independent of LPS stimulation. Although the calcium ionophore A23187 increased the expression of CD14 and TLR4, they did not exhibit energy transfer. However, following A23187 treatment, LPS promoted physical proximity between CD14 and TLR4. Therefore, we suggest that a close interaction between CD14 and TLR4 participates in LPS signaling, leading to nuclear translocation of NF-kappaB. PMID- 11034353 TI - NK cell inhibitory receptors prevent tyrosine phosphorylation of the activation receptor 2B4 (CD244). AB - 2B4 is an NK cell activation receptor that can provide a co-stimulatory signal to other activation receptors and whose mode of signal transduction is still unknown. We show that cross-linking of 2B4 on NK cells results in its rapid tyrosine phosphorylation, implying that this initial step in 2B4 signaling does not require coligation of other receptors. Ligation of 2B4 in the context of an NK cell-target cell interaction leads to 2B4 tyrosine phosphorylation, target cell lysis, and IFN-gamma release. Coligation of 2B4 with the inhibitory receptors killer cell Ig-like receptor (KIR)2DL1 or CD94/NKG2 completely blocks NK cell activation. The rapid tyrosine phosphorylation of 2B4 observed upon contact of NK cells with sensitive target cells is abrogated when KIR2DL1 or CD94/NKG2 are engaged by their cognate MHC class I ligand on resistant target cells. These results demonstrate that NK inhibitory receptors can interfere with a step as proximal as phosphorylation of an activation receptor. PMID- 11034354 TI - Defective NK cell activation in X-linked lymphoproliferative disease. AB - X-linked lymphoproliferative disease (XLP) is characterized by a selective immune deficiency to EBV. The molecular basis of XLP has been attributed to mutations of signaling lymphocytic activation molecule-associated protein, an intracellular molecule known to associate with the lymphocyte-activating surface receptors SLAM and 2B4. We have identified a single nucleotide mutation in SLAM-associated protein that affects the NK cell function of males carrying the mutated gene. In contrast to normal controls, both NK and lymphokine-activated killer cell cytotoxicity was significantly reduced in two XLP patients. In addition to decreased baseline cytotoxicity, ligation of 2B4 significantly augmented NK lytic function in normal controls but failed to enhance the cytotoxicity of NK cells from XLP patients. These findings suggest that association of SAP with 2B4 is necessary for optimal NK/lymphokine-activated killer cytotoxicity and imply that alterations in SAP/2B4 signaling contribute to the immune dysfunction observed in XLP. PMID- 11034355 TI - NK-mediated elimination of mutant lymphocytes that have lost expression of MHC class I molecules. AB - Mutant cells generated in vivo can be eliminated when mutated gene products are presented as altered MHC/peptide complexes and recognized by T cells. Diminished expression of MHC/peptide complexes enables mutant cells to escape recognition by T cells. In the present study, we tested the hypothesis that mutant lymphocytes lacking expression of MHC class I molecules are eliminated by autologous NK cells. In H-2b/k F1 mice, the frequency of H-2Kb-negative T cells was higher than that of H-2Kk-negative T cells. The frequency of H-2K-deficient T cells increased transiently after total body irradiation. During recovery from irradiation, H-2Kk negative T cells disappeared more rapidly than H-2Kb-negative T cells. The disappearance of H-2K-deficient T cells was inhibited by administration of Ab against asialo-GM1. H-2Kk-negative T cells showed higher sensitivity to autologous NK cells in vitro than H-2Kb/k heterozygous or H-2Kb-negative T cells. Adding syngeneic NK cells to in vitro cultures prevented emergence of mutant cells lacking H-2Kk expression but had little effect on the emergence of mutant cells lacking H-2Kb expression. Results in the H-2b/k F1 strain correspond with the sensitivity of parental H-2-homozygous cells in models of marrow graft rejection. In H-2b/d F1 mice, there was no significant difference between the frequencies of H-2Kb-negative and H-2Kd-negative T cells, although the frequencies of mutant cells were different after radiation exposure among the strains examined. H-2b/d F1 mice also showed rapid disappearance of the mutant T cells after irradiation, and administration of Ab against asialo-GM1 inhibited the disappearance of H-2K-deficient T cells in H-2b/d F1 mice. Our results provide direct evidence that autologous NK cells eliminate mutant cell populations that have lost expression of self-MHC class I molecules. PMID- 11034356 TI - Ligation of the WC1 receptor induces gamma delta T cell growth arrest through fumonisin B1-sensitive increases in cellular ceramide. AB - Ceramide is a powerful regulator of cell fate, inducing either apoptosis or growth arrest. We have previously shown that an Ab to the gammadelta T cell specific orphan receptor, WC1, is able to induce growth arrest in proliferating IL-2-dependent gammadelta T cells. We now show that this WC1-mediated growth arrest is associated with an increase in cellular ceramide, in the absence of any measurable changes in acidic/neutral sphingomyelinase activity. Moreover, cell permeable analogues of ceramide also mimicked WC1-induced growth arrest along with an associated decrease in pocket protein expression and phosphorylation status. An important role for ceramide in WC1-induced growth arrest was confirmed by demonstrating that the specific ceramide synthase inhibitor fumonisin B1 blocked WC1-induced growth arrest and the associated molecular effects on the pocket proteins. Finally, we observed constitutive expression of both antiapoptotic factors bcl-2 and bcl-X, the former having increased expression upon WC1 stimulation. It is therefore proposed that ligation of WC1 leads to an accumulation in cellular ceramide through activation of ceramide synthase. This in turn results in a decreased overall expression of the pocket proteins pRb and p107, their hypophosphorylation, and an eventual growth arrest of the gammadelta T cell. To our knowledge, these results demonstrate for the first time that cell surface receptor-mediated ceramide synthase activation can affect cell fate through increases in cellular ceramide and provide further evidence that the orphan receptor WC1 regulates gammadelta T cell biology through a novel signaling pathway. PMID- 11034357 TI - Distinct requirements for IFNs and STAT1 in NK cell function. AB - NK cell functions were examined in mice with a targeted mutation of the STAT1 gene, an essential mediator of IFN signaling. Mice deficient in STAT1 displayed impaired basal NK cytolytic activity in vitro and were unable to reject transplanted tumors in vivo, despite the presence of normal numbers of NK cells. IL-12 enhanced NK-mediated cytolysis, but poly(I:C) did not, and a similar phenotype occurred in mice lacking IFNalpha receptors. Molecules involved in activation and lytic function of NK cells (granzyme A, granzyme B, perforin, DAP10, and DAP12) were expressed at comparable levels in both wild-type and STAT1(-/-) mice, and serine esterase activity necessary for CTL function was normal, showing that the lytic machinery was intact. NK cells with normal cytolytic activity could be derived from STAT1(-/-) bone marrow progenitors in response to IL-15 in vitro, and enhanced NK lytic activity and normal levels of IFN-gamma were produced in response to IL-12 treatment in vivo. Despite these normal responses to cytokines, STAT1(-/-) mice could not reject the NK-sensitive tumor RMA-S, even following IL-12 treatment in vivo. Whereas in vitro NK cytolysis was also reduced in mice lacking both type I and type II IFN receptors, these mice resisted tumor challenge. These results demonstrate that both IFN alpha and IFN-gamma are required to maintain NK cell function and define a STAT1 dependent but partially IFN-independent pathway required for NK-mediated antitumor activity. PMID- 11034358 TI - Differential requirement of ZAP-70 for CD2-mediated activation pathways of mature human T cells. AB - This study addresses the role of the tyrosine kinase ZAP-70 in CD2-mediated T cell activation. Patients lacking ZAP-70 have few mature CD8+ T cells and high numbers of CD4+ T cells that are nonfunctional upon TCR triggering. Such a patient with a homozygous deletion in the zap-70 gene that resulted in the complete absence of ZAP-70 protein expression has been identified. Expression of the tyrosine kinases Lck, Fyn, and Syk was normal. The patient's T cells were activated with two different pairs of mitogenic mAbs. CD2-induced phosphorylation of the zeta-chain and influx of Ca2+ was defective in the ZAP-70-deficient T cells, whereas CD2-induced phosphorylation of several other proteins, including Syk, was not affected. CD2-induced proliferation as well as production of TNF alpha and IFN-gamma was abrogated in ZAP-70-deficient T cells, whereas PMA plus ionomycin induced normal activation of these cells. Together, this study shows that CD2-activation triggers ZAP-70-dependent and -independent pathways. Deletion of ZAP-70 affected CD2- and CD3-mediated proliferation and cytokine production in a similar way, suggesting that one of the different CD2 pathways converges with a CD3 pathway at or upstream of the activation of ZAP-70. PMID- 11034359 TI - Monocyte-derived CD1a+ and CD1a- dendritic cell subsets differ in their cytokine production profiles, susceptibilities to transfection, and capacities to direct Th cell differentiation. AB - We describe a phenotypically and functionally novel monocyte-derived dendritic cell (DC) subset, designated mDC2, that lacks IL-12 synthesis, produces high levels of IL-10, and directs differentiation of Th0/Th2 cells. Like conventional monocyte-derived DC, designated mDC1, mDC2 expressed high levels of CD11c, CD40, CD80, CD86, and MHC class II molecules. However, in contrast to mDC1, mDC2 lacked expression of CD1a, suggesting an association between cytokine production profile and CD1a expression in DC. mDC2 could be matured into CD83+ DC cells in the presence of anti-CD40 mAbs and LPS plus IFN-gamma, but they remained CD1a- and lacked IL-12 production even upon maturation. The lack of IL-12 and CD1a expression by mDC2 did not affect their APC capacity, because mDC2 stimulated MLR to a similar degree as mDC1. However, while mDC1 strongly favored Th1 differentiation, mDC2 directed differentiation of Th0/Th2 cells when cocultured with purified human peripheral blood T cells, further indicating functional differences between mDC1 and mDC2. Interestingly, the transfection efficiency of mDC2 with plasmid DNA vectors was significantly higher than that of mDC1, and therefore mDC2 may provide improved means to manipulate Ag-specific T cell responses after transfection ex vivo. Taken together, these data indicate that peripheral blood monocytes have the capacity to differentiate into DC subsets with different cytokine production profiles, which is associated with altered capacity to direct Th cell differentiation. PMID- 11034360 TI - Prolonged exposure to lipopolysaccharide inhibits macrophage 5-lipoxygenase metabolism via induction of nitric oxide synthesis. AB - LPS from bacteria can result in the development of sepsis syndrome and acute lung injury. Although acute exposure to endotoxin primes leukocytes for enhanced synthesis of leukotrienes (LT), little is known about the effect of chronic exposure. Therefore, we determined the effect of prolonged LPS treatment on 5 lipoxygenase (5-LO) metabolism of arachidonic acid in alveolar macrophages (AM) and in peripheral blood monocytes. Pretreatment of AM with LPS caused time- and dose-dependent suppression of LT synthetic capacity. LPS pretreatment failed to inhibit arachidonic acid (AA) release. The fact that LPS inhibited LT synthesis from endogenous AA more than from exogenous AA suggested an effect on 5-LO activating protein (FLAP). In addition, an inhibitory effect of LPS treatment on AM 5-LO activity was suggested by cell-free 5-LO enzyme assay. No effect on the expression of either 5-LO or FLAP proteins was observed. New protein synthesis was necessary for LPS-induced reduction of 5-LO metabolism in AM, and immunoblotting demonstrated marked induction of NO synthase (NOS). Inhibition by LPS was reproduced by an NO donor and was abrogated by inhibitors of constitutive and inducible NOS. Compared with AM, peripheral blood monocytes exhibited no suppression by LPS of 5-LO metabolism and no induction of inducible NOS. We conclude that prolonged exposure to LPS impairs AM 5-LO metabolism by NO-mediated suppression of both 5-LO and FLAP function. Because LT contribute to antimicrobial defense, this down-regulation of 5-LO metabolism may contribute to the increased susceptibility to pneumonia in patients following sepsis. PMID- 11034361 TI - Role of tolerogen conformation in induction of oral tolerance in experimental autoimmune myasthenia gravis. AB - We recently demonstrated that oral or nasal administration of recombinant fragments of the acetylcholine receptor (AChR) prevents the induction of experimental autoimmune myasthenia gravis (EAMG) and suppresses ongoing EAMG in rats. We have now studied the role of spatial conformation of these recombinant fragments in determining their tolerogenicity. Two fragments corresponding to the extracellular domain of the human AChR alpha-subunit and differing in conformation were tested: Halpha1-205 expressed with no fusion partner and Halpha1-210 fused to thioredoxin (Trx), and designated Trx-Halpha1-210. The conformational similarity of the fragments to intact AChR was assessed by their reactivity with alpha-bungarotoxin and with anti-AChR mAbs, specific for conformation-dependent epitopes. Oral administration of the more native fragment, Trx-Halpha1-210, at the acute phase of disease led to exacerbation of EAMG, accompanied by an elevation of AChR-specific humoral and cellular reactivity, increased levels of Th1-type cytokines (IL-2, IL-12), decreased levels of Th2 (IL 10)- or Th3 (TGF-beta)-type cytokines, and higher expression of costimulatory factors (CD28, CTLA4, B7-1, B7-2, CD40L, and CD40). On the other hand, oral administration of the less native fragments Halpha1-205 or denatured Trx-Halpha1 210 suppressed ongoing EAMG and led to opposite changes in the immunological parameters. It thus seems that native conformation of AChR-derived fragments renders them immunogenic and immunopathogenic and therefore not suitable for treatment of myasthenia gravis. Conformation of tolerogens should therefore be given careful attention when considering oral tolerance for treatment of autoimmune diseases. PMID- 11034362 TI - Mutant mouse lysozyme carrying a minimal T cell epitope of hen egg lysozyme evokes high autoantibody response. AB - Self proteins including foreign T cell epitope induce autoantibodies. We evaluated the relationship between the size of foreign Ag introduced into self protein and the magnitude of autoantibody production. Mouse lysozyme (ML) was used as a model self protein, and we prepared three different ML derivatives carrying T cell epitope of hen egg white lysozyme (HEL) 107-116, i.e, heterodimer of ML and HEL (ML-HEL), chimeric lysozyme that has residue 1-82 of ML and residue 83-130 of HEL in its sequence (chiMH), and mutant ML that has triple mutations rendering the most potent T cell epitope of HEL (sequence 107-116). Immunization of BALB/c mice with these three ML derivatives induced anti-ML autoantibody responses, whereas native ML induced no detectable response. In particular, mutML generated a 10(4) times higher autoantibody titer than did ML-HEL. Anti-HEL107 116 T cell-priming activities were almost similar among the ML derivatives. The heterodimerization of mutant ML and HEL led to significant reduction of the autoantibody response, whereas the mixture did not. These results show that size of the nonself region in modified self Ag has an important role in determining the magnitude of the autoantibody response, and that decrease in the foreign region in a modified self protein may cause high-titered autoantibody response. PMID- 11034364 TI - Conventional, naive CD4+ T cells provide an initial source of IL-4 during Th2 differentiation. AB - IL-4 is known to promote the differentiation of CD4+ T cells into IL-4-secreting Th2 cells. However, the cellular source of the early burst of IL-4 that drives Th2 responses in vivo has not been conclusively identified. Mice deficient for the IL-4 receptor alpha-chain (IL-4Ralpha-/-) retain the capacity to secrete IL-4 and can be used to identify those cell types that produce IL-4 without a requirement for prior IL-4-mediated stimulation. To address whether naive, conventional CD4+ T cells may act as initial producers of IL-4 in Ag-specific responses, we crossed the BALB/c IL-4Ralpha-/-mice to DO11.10/scid TCR transgenic mice. Lymph node cells from wild-type and IL-4Ralpha-/- DO11.10/scid mice secreted approximately 50 pg of IL-4 per10(6) cells within 48 h after peptide stimulation. This small amount of IL-4 was sufficient to cause the differentiation of wild-type CD4+ T cells into Th2 cells, particularly if IFN gamma and IL-12 were neutralized during the priming cultures. CD4+ cells from the IL-4Ralpha-/- mice gave rise to a minor proportion (approximately 2%) of IL-4 producing cells upon stimulation in the presence of anti-IFN-gamma and anti-IL 12. These data show that conventional, naive CD4+ T cells may be considered as initial sources of IL-4 and, in the absence of IFN-gamma and IL-12, this IL-4 can induce Th2 polarization. PMID- 11034363 TI - CD4 help-independent induction of cytotoxic CD8 cells to allogeneic P815 tumor cells is absolutely dependent on costimulation. AB - Mice made transgenic (Tg) for a rat anti-mouse CD4 Ab (GK mice) represent a novel CD4-deficient model. They not only lack canonical CD4 cells in the periphery, but also lack the residual aberrant Th cells that are found in CD4-/- mice and MHC class II-/- mice. To analyze the role of CD4 help and costimulation for CTL induction against alloantigens, we have assessed the surface and functional phenotype of CD8 cells in vivo (e.g., clearance of allogeneic P815 cells) and in vitro. In our CD4-deficient GK mice, CTL responses to allogeneic P815 cells were induced, albeit delayed, and were sufficient to eliminate P815 cells. Induction of CTL and elimination of allogeneic P815 cells were inhibited both in the presence and absence of CD4 cells by temporary CD40 ligand blockade. This indicated that direct interaction of CD40/CD40L between APCs and CD8 cells may be an accessory signal in CTL induction (as well as the indirect pathway via APC/CD4 interaction). Furthermore, whereas in CTLA4Ig single Tg mice P815 cells were rejected promptly, in the double Tg GK/CTLA4Ig mice CTL were not induced and allogeneic P815 cells were not rejected. These findings suggest that CD40/CD40L is involved in both CD4-dependent and CD4-independent pathways, and that B7/CD28 is pivotal in the CD4-independent pathway of CTL induction against allogeneic P815 cells. PMID- 11034365 TI - IL-10, but not IL-4, suppresses infection-stimulated bone resorption in vivo. AB - Periapical bone resorption occurs following infection of the dental pulp and is mediated mainly by IL-1alpha in the murine model. The production and activity of IL-1alpha is modulated by a network of regulatory cytokines, including those produced by Th1 (pro-inflammatory) and Th2 (anti-inflammatory) subset T cells. This study was designed to assess the functional role of the Th2-type cytokines IL-4 and IL-10 in infection-stimulated bone resorption in vivo. The dental pulps of the first molars were exposed and infected with a mixture of four common endodontic pathogens, and bone destruction was determined by micro-computed tomography at sacrifice on day 21. The results demonstrate that IL-10(-/-) mice had significantly greater infection-stimulated bone resorption in vivo compared with wild-type mice (p < 0.001), whereas IL-4(-/-) exhibited no increased resorption. IL-10(-/-) had markedly elevated IL-1alpha production within periapical inflammatory tissues (>10-fold) compared with wild type (p < 0.01), whereas IL-4(-/-) exhibited decreased IL-1alpha production (p < 0.05). IL-10 also suppressed IL-1alpha production by macrophages in a dose-dependent fashion in vitro, whereas IL-4 had weak and variable effects. We conclude that IL-10, but not IL-4, is an important endogenous suppressor of infection-stimulated bone resorption in vivo, likely acting via inhibition of IL-1alpha expression. PMID- 11034366 TI - Effects of a hexameric deoxyriboguanosine run conjugation into CpG oligodeoxynucleotides on their immunostimulatory potentials. AB - CpG oligodeoxynucleotides (ODNs) are promising immunomodulatory agents for treating human diseases and vaccine development. Phosphodiester CpG ODNs were demonstrated to have poor immunostimulatory potentials for cytokine production. However, the conjugation of consecutive deoxyriboguanosine residues, called a dG run, at the 3' terminus of phosphodiester CpG ODNs significantly enhanced TNF alpha and IL-12 production from mouse splenic dendritic cells (DCs). The optimal induction of cytokine production was achieved by the addition of a hexameric dG (dG6) run. In contrast, the existence of a dG6 run either at the 5' terminus of phosphodiester CpG ODNs or at the 3' terminus of phosphorothioate CpG ODNs diminished CpG-mediated cytokine induction, suggesting that the effects of a dG run depend on its location and the chemical property of the ODN backbone, respectively. In addition, we provided the evidence that the conjugation of a dG6 run caused the structural transformation of CpG ODNs, which facilitates their targeting into mouse APCs such as splenic DCs, B cells, and peritoneal macrophages with a scavenger receptor type A ligand specificity. Among primary APCs, DCs were the most potent for CpG ODN-mediated IL-12 production. Furthermore, we demonstrated that the conjugation of a dG6 run into the 3' terminus of phosphodiester CpG ODNs was crucial for their ability to generate Th1 immunity in vivo. Thus, the conjugation of a dG6 run into phosphodiester CpG ODNs would be an alternative way to optimize their immunostimulatory potentials in vitro and in vivo. PMID- 11034367 TI - Cellular interactions involved in Th cell memory. AB - The cellular interactions involved in maintaining CD4+ T cell memory have hitherto not been identified. In this report, we have investigated the roles played by B cells and dendritic cells (DCs) in this process. We show that long lasting Th cell memory depends on the presence of B cells, but that direct Ag presentation by B cells is not required. Instead, Ag presentation by DCs is critical for the survival of memory Th cells. DCs presenting specific Ag can be detected in animals long after immunization. These findings support a model in which B cells provide an environment in which Ags may be trapped and retained. This Ag is periodically presented to memory CD4+ T cells by DCs, providing an essential survival signal. PMID- 11034368 TI - Bacterial lipopolysaccharide, TNF-alpha, and calcium ionophore under serum-free conditions promote rapid dendritic cell-like differentiation in CD14+ monocytes through distinct pathways that activate NK-kappa B. AB - To facilitate the study of signaling pathways involved in myeloid dendritic cell (DC) differentiation, we have developed a serum-free culture system in which human CD14+ peripheral blood monocytes differentiate rapidly in response to bacterial LPS, TNF-alpha, or calcium ionophore (CI). Within 48-96 h, depending on the inducing agent, the cells acquire many immunophenotypical, morphological, functional, and molecular properties of DC. However, there are significant differences in the signaling pathways used by these agents, because 1) LPS induced, but not CI-induced, DC differentiation required TNF-alpha production; and 2) cyclosporin A inhibited differentiation induced by CI, but not that induced by LPS. Nevertheless, all three inducing agents activated members of the NF-kappaB family of transcription factors, including RelB, suggesting that despite differences in upstream elements, the signaling pathways all involve NF kappaB. In this report we also demonstrate and offer an explanation for two observed forms of the RelB protein and show that RelB can be induced in myeloid cells, either directly or indirectly, through a calcium-dependent and cyclosporin A-sensitive pathway. PMID- 11034369 TI - Cross-presentation of tumor antigens to effector T cells is sufficient to mediate effective immunotherapy of established intracranial tumors. AB - The systemic adoptive transfer of tumor-sensitized T cells, activated ex vivo, can eliminate established intracranial tumors. Regression of MHC class II negative MCA 205 fibrosarcomas occurs optimally following adoptive transfer of both CD4 and CD8 tumor-sensitized T cells, indicating an important function for tumor-infiltrating APC. Here, we demonstrate that during an effector response, indirect presentation of tumor Ags to transferred T cells is sufficient to mediate intracranial tumor regression. BALB/c --> CB6F1 (H-2bxd) bone marrow chimeras were challenged with the MCA 205 fibrosarcoma (H-2b). The tumor grew progressively in the H-2b-tolerant chimeras and stimulated an immune response in tumor-draining lymph nodes. Tumor-sensitized lymph node T cells were activated ex vivo with anti-CD3 and IL-2, then adoptively transferred to sublethally irradiated BALB/c or C57BL/6 recipients bearing established intracranial MCA 205 tumors. The transferred T cells eradicated MCA 205 tumors in BALB/c recipients and demonstrated tumor specificity, but had no therapeutic efficacy in the C57BL/6 recipients. These data establish that tumor-associated host cell constituents provide sufficient Ag presentation to drive effector T cell function in the complete absence of direct tumor recognition. This effector mechanism has an evident capacity to remain operative in circumstances of immune escape, where the tumor does not express the relevant MHC molecules, and may have importance even at times when direct CTL recognition also remains operative. PMID- 11034370 TI - Cytotoxic T cells specifically induce Fas on target cells, thereby facilitating exocytosis-independent induction of apoptosis. AB - Cytotoxic T (Tc) cells deficient in perforin lyse Fas-negative targets after lengthy incubation periods. This process is independent of granzymes, and killing occurs via the Fas pathway for the following reasons. Interaction of perforin deficient Tc cells with Fas-negative targets leads to an up-regulation of Fas that is dependent on Ag recognition, de novo synthesis, and transport of proteins to the target cell surface. Treatment of effectors with brefeldin A, but not with the exocytosis inhibitor concanamycin, inhibited this process. Lysis of targets is inhibited by anti-Fas Abs, soluble mouse Fas-Fc, and the caspase-cascade inhibitor, crm-A. Targets from Fas-mutant lpr mice are refractory to lysis, and Tc cells from mice deficient in Fas- and perforin-mediated lysis do not lyse Fas negative targets. The possible relevance of this exocytosis-independent cytolytic process in the regulation of T cell activity and control of pathogens is discussed. PMID- 11034371 TI - CD8+ T cells rapidly acquire NK1.1 and NK cell-associated molecules upon stimulation in vitro and in vivo. AB - NKT cells express both NK cell-associated markers and TCR. Classically, these NK1.1+TCRalphabeta+ cells have been described as being either CD4+CD8- or CD4-CD8 . Most NKT cells interact with the nonclassical MHC class I molecule CD1 through a largely invariant Valpha14-Jalpha281 TCR chain in conjunction with either a Vbeta2, -7, or -8 TCR chain. In the present study, we describe the presence of significant numbers of NK1.1+TCRalphabeta+ cells within lymphokine-activated killer cell cultures from wild-type C57BL/6, CD1d1-/-, and Jalpha281-/- mice that lack classical NKT cells. Unlike classical NKT cells, 50-60% of these NK1.1+TCRalphabeta+ cells express CD8 and have a diverse TCR Vbeta repertoire. Purified NK1.1-CD8alpha+ T cells from the spleens of B6 mice, upon stimulation with IL-2, IL-4, or IL-15 in vitro, rapidly acquire surface expression of NK1.1. Many NK1.1+CD8+ T cells had also acquired expression of Ly-49 receptors and other NK cell-associated molecules. The acquisition of NK1.1 expression on CD8+ T cells was a particular property of the IL-2Rbeta+ subpopulation of the CD8+ T cells. Efficient NK1.1 expression on CD8+ T cells required Lck but not Fyn. The induction of NK1.1 on CD8+ T cells was not just an in vitro phenomenon as we observed a 5-fold increase of NK1.1+CD8+ T cells in the lungs of influenza virus infected mice. These data suggest that CD8+ T cells can acquire NK1.1 and other NK cell-associated molecules upon appropriate stimulation in vitro and in vivo. PMID- 11034372 TI - A profound deficiency in thymic progenitor cells in mice lacking Jak3. AB - Humans and mice with genetic deficiencies that lead to loss of signaling through common gamma-chain (gammac)-containing cytokine receptors have severe defects in B and T lymphocytes. In humans, these deficiencies lead to a complete absence of T cells, whereas in mice, small thymuses give rise to normal numbers of peripheral T cells. We have examined the first wave of developing T cells in Jak3 /-, IL-7-/-, and IL-7Ralpha-/- fetal mice, and have found a near absence of thymic progenitor cells. This deficiency is highlighted by the complete inability of Jak3-/- progenitor cells to reconstitute T cell development in the presence of competing wild-type cells. These data clearly demonstrate a strong common basis for the T cell deficiencies in mice and humans lacking gammac/Jak3 signaling pathways. PMID- 11034373 TI - Differential survival of naive CD4 and CD8 T cells. AB - In this paper we compare survival characteristics of transgenic and polyclonal CD4 and CD8 T cells. Transgenic CD4 T cells have an intrinsically lower capacity for survival, reflected in their gradual disappearance in thymectomized hosts, their increased sensitivity to apoptosis in vitro, and fewer divisions during homeostatic proliferation upon transfer into syngeneic lymphopenic hosts compared with CD8 T cells. Homeostatic proliferation, however, does not generally result in phenotypic conversion of activation markers unless cognate or cross-reactive Ag is present. T cells from the A18 TCR transgenic strain normally selected into the CD4 lineage are fragile as CD4 T cells, yet display the typical robust survival pattern of CD8 T cells when diverted into the CD8 lineage in a CD4 deficient host. Polyclonal CD4 and CD8 T cells also show distinctive patterns of survival, emphasizing that survival signals are relayed differently in the two lymphocyte subpopulations. However, expression levels of Bcl-2 in either transgenic or polyclonal naive CD4 and CD8 T cells are similar, excluding a role for this molecule as a key factor in differential survival of CD4 vs CD8 T cells. PMID- 11034374 TI - Development of dendritic epidermal T cells with a skewed diversity of gamma delta TCRs in V delta 1-deficient mice. AB - One of the most intriguing features of gammadelta T cells that reside in murine epithelia is the association of a specific Vgamma/Vdelta usage with each epithelial tissue. Dendritic epidermal T cells (DETCs) in the murine epidermis, are predominantly derived from the "first wave" Vgamma5+ fetal thymocytes and overwhelmingly express the canonical Vgamma5/Vdelta1-TCRs lacking junctional diversity. Targeted disruption of the Vdelta1 gene resulted in a markedly impaired development of Vgamma5+ fetal thymocytes as precursors of DETCs; however, gammadeltaTCR+ DETCs with a typical dendritic morphology were observed in Vdelta1-/- mice and their cell densities in the epidermis were slightly lower than those in Vdelta1+/- epidermis. Moreover, the Vdelta1-deficient DETCs were functionally competent in their ability to up-regulate cytokines and keratinocyte growth factor-expression in response to keratinocytes. Vgamma5+ DETCs were predominant in the Vdelta1-/- epidermis, though Vgamma5- gammadeltaTCR+ DETCs were also detected. The Vgamma5+ DETCs showed a typical dendritic shape, gammadeltaTCR(high), and age-associated expansion in epidermis as observed in conventional DETCs of normal mice, whereas the Vgamma5- gammadeltaTCR+ DETCs showed a less dendritic shape, gammadeltaTCR(low), and no expansion in the epidermis, consistent with their immaturity. These results suggest that optimal DETC development does not require a particular Vgamma/Vdelta-chain usage but requires expression of a limited diversity of gammadeltaTCRs, which allow DETC precursors to mature and expand within the epidermal microenvironment. PMID- 11034375 TI - Thymocyte maturation: selection for in-frame TCR alpha-chain rearrangement is followed by selection for shorter TCR beta-chain complementarity-determining region 3. AB - Thymocyte maturation consists of a number of stages, the goal of which is the production of functioning T cells that respond to foreign antigenic peptides using their clonotypic receptors. Selection of a productively rearranged TCR beta chain is the first stage in the process and occurs at the double-negative to double-positive (DP) transition. Later maturation stages are based on changes in markers such as CD5, CD69, or IL-7R. A stage in which a-chains are selected has also been identified using beta-chain transgenic mice. Here we identify two additional selection stages in human thymocytes based on characteristics of the TCR. alpha selection is measured directly by identification of in-frame rearrangements and is associated with the appearance of CD3 on the DP thymocyte surface. The next stage has not yet been described and involves selection of thymocytes that express shorter TCR beta-chain complementarity-determining region 3 (CDR3). This stage is associated with the acquisition of high levels of CDR3 by DP cells and the transition to SP thymocytes. The extent of CDR3 length selection observed is a function of the TCR V and J genes. We propose that CDR3 length selection is based on recognition of the MHC. Thus, there exist limitations on the allowable length of that portion of the TCR most intimately in contact with MHC and peptide. This may be a physical representation of positive selection. PMID- 11034376 TI - Effects of a constitutively active form of calcineurin on T cell activation and thymic selection. AB - Calcineurin is a calcium/calmodulin-dependent phosphatase whose activity is required for the induction of T cell lymphokine production and proliferation. Although its specific role in T cell development is less well defined, studies with the immunosuppressive drugs cyclosporin A and FK-506 suggest that it is involved in both positive and negative selection of immature thymocytes. To more completely characterize a role for calcineurin in T cell development in vivo, we have generated transgenic mice that express an activated form of this enzyme in thymocytes and peripheral T cells. We find that the transgene causes a block in early thymic development, resulting in a reduction in the steady-state number of CD4 and CD8 double positives, but not on the number of mature T cells. We also find that thymocytes and mature T cells expressing this transgene are more sensitive to signals through their TCR. In thymocytes this sensitivity difference is manifested as an increase in positive selection, although negative selection seems to remain unaffected. Therefore, these studies confirm and extend past reports that suggested a role for calcineurin in thymic development and selection. PMID- 11034378 TI - The mechanism of unresponsiveness to circulating tumor antigen MUC1 is a block in intracellular sorting and processing by dendritic cells. AB - Immunity to tumor Ags in patients is typically weak and not therapeutic. We have identified a new mechanism by which potentially immunogenic glycoprotein tumor Ags, such as MUC1, fail to stimulate strong immune responses. MUC1 is a heavily glycosylated membrane protein that is also present in soluble form in sera and ascites of cancer patients. We show that this soluble protein is readily taken up by dendritic cells (DC), but is not transported to late endosomes or MHC class II compartments for processing and binding to class II MHC. MUC1 uptake is mediated by the mannose receptor, and the protein is then retained long term in early endosomes without degradation. Long-term retention of MUC1 does not interfere with the ability of DC to process and present other Ags. We also demonstrate inhibited processing of another important glycoprotein tumor Ag, HER-2/neu. This may, therefore, be a frequent obstacle to presentation of tumor Ags and an important consideration in the design of cancer vaccines. It should be possible to overcome this obstacle by providing DC with a form of tumor Ag that can be better processed. For MUC1 we show that a 140-aa-long synthetic peptide is very efficiently processed by DC. PMID- 11034377 TI - Galectin-1 induces partial TCR zeta-chain phosphorylation and antagonizes processive TCR signal transduction. AB - Galectin-1 is an endogenous lectin with known T cell immunoregulatory activity, though the molecular basis by which galectin-1 influences Ag specific T cell responses has not been elucidated. Here, we characterize the ability of galectin 1 to modulate TCR signals and responses by T cells with well defined hierarchies of threshold requirements for signaling distinct functional responses. We demonstrate that galectin-1 antagonizes TCR responses known to require costimulation and processive protein tyrosine phosphorylation, such as IL-2 production, but is permissive for TCR responses that only require partial TCR signals, such as IFN-gamma production, CD69 up-regulation, and apoptosis. Galectin-1 binding alone or together with Ag stimulation induces partial phosphorylation of TCR-zeta and the generation of inhibitory pp21zeta. Galectin-1 antagonizes Ag induced signals and TCR/costimulator dependent lipid raft clustering at the TCR contact site. We propose that galectin-1 functions as a T cell "counterstimulator" to limit required protein segregation and lipid raft reorganization at the TCR contact site and, thus, processive and sustained TCR signal transduction. These findings support the concept that TCR antagonism can arise from the generation of an inhibitory pp21zeta-based TCR signaling complex. Moreover, they demonstrate that TCR antagonism can result from T cell interactions with a ligand other than peptide/MHC. PMID- 11034379 TI - The CD85/LIR-1/ILT2 inhibitory receptor is expressed by all human T lymphocytes and down-regulates their functions. AB - The inhibitory molecule CD85/LIR-1/ILT2 has been detected previously on the surface of a small proportion of T lymphocytes. In this study, evidence is provided that, although only a fraction of CD3+ cells are stained by mAb specific for CD85/LIR-1/ILT2 on their surface, this inhibitory receptor is present in the cytoplasm of all T lymphocytes, and that it is detectable on the surface of all T cell clones by the M402 mAb. Biochemical analyses further demonstrate that CD85/LIR-1/ILT2 is present in all T clones analyzed, and that the protein is tyrosine-phosphorylated. Expression of mRNA coding for CD85/LIR-1/ILT2 has been assessed by RT-PCR. Notably, in the NKL cell line and in one T cell clone, amplification of the messenger required 30 cycles only, whereas, in other T cell clones, an amplification product was detected by increasing the number of cycles. CD85/LIR-1/ILT2 inhibits CD3/TCR-mediated activation in both CD4+ and CD8+ clones, and it down-regulates Ag recognition by CD8+ cells in a clonally distributed fashion. Addition of anti-ILT2 HP-F1 mAb in the cytolytic assay enhances target cell lysis mediated by Ag-specific CTL. This could be due to interference of the mAb with receptor/ligand interactions. In contrast, HP-F1 mAb cross-linking triggers inhibitory signals that reduce cytotoxicity. CD85/LIR 1/ILT2 also controls responses to recall Ags and, in low responders, its engagement sharply increases T cell proliferation. The inhibitory function of the molecule is also confirmed by its ability to reduce CD3/TCR-induced intracellular Ca2+ mobilization. PMID- 11034380 TI - CD1d on myeloid dendritic cells stimulates cytokine secretion from and cytolytic activity of V alpha 24J alpha Q T cells: a feedback mechanism for immune regulation. AB - The precise immunologic functions of CD1d-restricted, CD161+ AV24AJ18 (Valpha24JalphaQ) T cells are not well defined, although production of IL-4 has been suggested as important for priming Th2 responses. However, activation of human Valpha24JalphaQ T cell clones by anti-CD3 resulted in the secretion of multiple cytokines notably important for the recruitment and differentiation of myeloid dendritic cells. Specific activation of Valpha24JalphaQ T cells was CD1d restricted. Expression of CD1d was found on monocyte-derived dendritic cells in vitro, and immunohistochemical staining directly revealed CD1d preferentially expressed on dendritic cells in the paracortical T cell zones of lymph nodes. Moreover, myeloid dendritic cells both activated Valpha24JalphaQ T cells and were susceptible to lysis by these same regulatory T cells. Because myeloid dendritic cells are a major source of IL-12 and control Th1 cell differentiation, their elimination by lysis is a mechanism for limiting the generation of Th1 cells and thus regulating Th1/Th2 responses. PMID- 11034381 TI - A monoclonal antibody against the 66-kDa protein expressed in mouse spleen and thymus inhibits Ly-6A.2-dependent cell-cell adhesion. AB - The Ly-6 locus encodes several cell surface proteins of 10-12 kDa. Some members of this multigene family may function in cell signaling and/or cell adhesion processes. T lymphocytes overexpressing Ly-6A.2 (one member of the Ly-6 gene family) protein homotypically aggregate when cultured in vitro. Further analysis of this homotypic aggregation suggests that Ly-6A.2 participates in cell-cell adhesion. These observations indicated the presence of a Ly-6 ligand(s) on the surface of lymphoid cells. In this study we report generation of a hamster mAb, 9AB2, that blocks Ly-6A.2-dependent cell-cell adhesion. The 9AB2 Ab recognizes a 66-kDa glycoprotein with unique tissue expression. The 9AB2 mAb does not bind Ly 6A.2, but coimmunoprecipitates Ly-6A.2 molecule. Moreover, 9AB2 Ag-expressing thymocytes specifically bind to Chinese hamster ovary cells overexpressing Ly 6A.2 protein, and this binding is specifically blocked by 9AB2 and anti-Ly-6A.2 Abs. These results suggest that the 66-kDa protein recognized by 9AB2 mAb is the putative ligand for Ly-6A.2. PMID- 11034382 TI - Plasmid vaccine expressing granulocyte-macrophage colony-stimulating factor attracts infiltrates including immature dendritic cells into injected muscles. AB - Plasmid-encoded GM-CSF (pGM-CSF) is an adjuvant for genetic vaccines; however, little is known about how pGM-CSF enhances immunogenicity. We now report that pGM CSF injected into mouse muscle leads to a local infiltration of potential APCs. Infiltrates reached maximal size on days 3 to 5 after injection and appeared in several large discrete clusters within the muscle. Immunohistological studies in muscle sections from mice injected with pGM-CSF showed staining of cells with the macrophage markers CD11b, Mac-3, IA(d)/E(d) and to the granulocyte marker GR-1 from day 1 through day 14. Cells staining with the dendritic cell marker CD11c were detected only on days 3 to 5. Muscles injected with control plasmids did not stain for CD11c but did stain for CD11b, Mac-3, IA(d)/E(d), and GR-1. No staining was observed with the APC activation markers, B7.1 or CD40, or with markers for T or B cells. These findings are consistent with the infiltrating cells in the pGM CSF-injected muscles being a mixture of neutrophils, macrophages, and immature dendritic cells and suggest that the i.m. APCs may be enhancing immune responses to coinjected plasmid Ags. This hypothesis is supported by data showing that 1) separation of injections with pGM-CSF and Ag-expressing plasmid into different sites did not enhance immune responses and 2) immune enhancement was associated with the presence of CD11c+ cells in the infiltrates. Thus, pGM-CSF enhancement may depend on APC recruitment to the i.m. site of injection. PMID- 11034383 TI - Type 1 IFN maintains the survival of anergic CD4+ T cells. AB - Anergic T cells have immunoregulatory activity and can survive for extended periods in vivo. It is unclear how anergic T cells escape from deletion, because both anergy and apoptosis can occur after TCR ligation. Stimulation of human CD4+ T cell clones reactive to influenza hemagglutinin peptides can occur in the absence of APCs when MHC class II-expressing, activated T cells present peptide to each other. This T:T peptide presentation can induce CD95-mediated apoptosis, while the cells that do not die are anergic. We found that the death after peptide or anti-CD3 treatment of a panel of CD4+ T cell clones is blocked by IFN beta secreted by fibroblasts and also by IFN-alpha. This increases cell recovery after stimulation, which is not due to T cell proliferation. This mechanism for apoptosis inhibition rapidly stops protein kinase C-delta translocation from the cytoplasm to the nucleus, which is an early event in the death process. A central observation was that CD4+ T cells that are rescued from apoptosis after T:T presentation of peptide by IFN-alphabeta remain profoundly anergic to rechallenge with Ag-pulsed APCs. However, anergized cells retain the ability to respond to IL 2, showing that they are nonresponsive but functional. The prevention of peptide induced apoptosis in activated T cells by IFN-alphabeta is a novel mechanism that may enable the survival and maintenance of anergic T cell populations after TCR engagement. This has important implications for the persistence of anergic T cells with the potential for immunoregulatory function in vivo. PMID- 11034384 TI - The role of the DAP12 signal in mouse myeloid differentiation. AB - DAP12 is a recently cloned, immunoreceptor tyrosine-based activation motif bearing transmembrane adapter molecule that is associated with the NK-activating receptors. Previous reports showed that the DAP12 message could be detected not only in NK cells but also in granulocytes, monocytes, dendritic cells, and macrophages. In this study we found a significant level of DAP12 protein expression in macrophage-related cell lines and organs. Additionally, we observed increased expression of DAP12 after LPS-induced differentiation of M1 cells into macrophages. To examine the role of DAP12 in the myeloid cell lineage, we established M1 FLAG-DAP12 transfectants (FDAP-M1) and demonstrated the marked morphological changes in FDAP-M1 cells caused by signaling through DAP12. Cell surface phenotypic analysis showed up-regulation of macrophage markers CD11b, 2.4G2, and adhesion molecule B7-2. Additionally, after stimulation through DAP12, phosphorylated FLAG -DAP12 could be immunoprecipitated using anti-phosphotyrosine mAbs. Collectively, these findings indicate that direct DAP12 signaling has an important role in macrophage differentiation. PMID- 11034385 TI - Dendritic cells capture killed tumor cells and present their antigens to elicit tumor-specific immune responses. AB - Due to their capacity to induce primary immune responses, dendritic cells (DC) are attractive vectors for immunotherapy of cancer. Yet the targeting of tumor Ags to DC remains a challenge. Here we show that immature human monocyte-derived DC capture various killed tumor cells, including Jurkat T cell lymphoma, malignant melanoma, and prostate carcinoma. DC loaded with killed tumor cells induce MHC class I- and class II-restricted proliferation of autologous CD8+ and CD4+ T cells, demonstrating cross-presentation of tumor cell-derived Ags. Furthermore, tumor-loaded DC elicit expansion of CTL with cytotoxic activity against the tumor cells used for immunization. CTL elicited by DC loaded with the PC3 prostate carcinoma cell bodies kill another prostate carcinoma cell line, DU145, suggesting recognition of shared Ags. Finally, CTL elicited by DC loaded with killed LNCap prostate carcinoma cells, which express prostate specific Ag (PSA), are able to kill PSA peptide-pulsed T2 cells. This demonstrates that induced CTL activity is not only due to alloantigens, and that alloantigens do not prevent the activation of T cells specific for tumor-associated Ags. This approach opens the possibility of using allogeneic tumor cells as a source of tumor Ag for antitumor therapies. PMID- 11034386 TI - Activation of toll-like receptor 2 on human dendritic cells triggers induction of IL-12, but not IL-10. AB - Mammalian Toll-like receptors (TLRs) are required for cell activation by bacterial lipoproteins (bLP) and LPS. Stimulation of monocytes with bLP and LPS results in a TLR-dependent induction of immunomodulatory genes leading to the production of pro-inflammatory cytokines. In this paper, we compared the expression and response of TLRs on monocytes and dendritic cells (DC). TLR2, but not TLR4, was detected on peripheral blood monocytes and DC, in lymphoid tissue CD1alpha+ DC as well as on in vitro monocyte-derived DC. Upon stimulation with bLP or LPS, monocytes produced IL-12 and IL-10 at similar levels, whereas monocyte-derived DC produced comparable levels of IL-12, but little IL-10. Greater than 90% of the bLP-induced production of IL-12 was blocked by anti-TLR2 mAb. Thus, DC express TLR2 and activation of this receptor by bLP provides an innate mechanism by which microbial pathogens preferentially activate cell mediated immunity. PMID- 11034387 TI - Direct tumor lysis by NK cells uses a Ras-independent mitogen-activated protein kinase signal pathway. AB - Destruction of tumor cells is a key function of lymphocytes, but the molecular processes driving it are unclear. Analysis of signal molecules indicated that mitogen-activated protein kinase (MAPK)/extracellular regulated kinase 2 critically controlled lytic function in human NK cells. We now have evidence to indicate that target ligation triggers a Ras-independent MAPK pathway that is required for lysis of the ligated tumor cell. Target engagement caused NK cells to rapidly activate MAPK within 5 min, and PD098059 effectively blocked both MAPK activation and tumoricidal function in NK cells. Target engagement also rapidly activated Ras, detected as active Ras-GTP bound to GST-Raf-RBD, a GST fusion protein linked to the Raf protein fragment containing the Ras-GTP binding domain. However, Ras inactivation by pharmacological disruption with the farnesyl transferase inhibitor, FTI-277, had no adverse effect on the ability of NK cells to lyse tumor cells or to express MAPK activation upon target conjugation. Notably, MAPK inactivation with PD098059, but not Ras inactivation with FTI-277, could interfere with perforin and granzyme B polarization within NK cells toward the contacted target cell. Using vaccinia delivery of N17 Ras into NK cells, we demonstrated that IL-2 activated a Ras-dependent MAPK pathway, while target ligation used a Ras-independent MAPK pathway to trigger lysis in NK cells. PMID- 11034388 TI - CD28 utilizes Vav-1 to enhance TCR-proximal signaling and NF-AT activation. AB - The mechanism through which CD28 costimulation potentiates TCR-driven gene expression is still not clearly defined. Vav-1, an exchange factor for Rho GTPases thought to regulate, mainly through Rac-1, various signaling components leading to cytokine gene expression, is tyrosine phosphorylated upon CD28 engagement. Here, we provide evidence for a key role of Vav-1 in CD28-mediated signaling. Overexpression of Vav-1 in Jurkat cells in combination with CD28 ligation strongly reduced the concentration of staphylococcus enterotoxin E/MHC required for TCR-induced NF-AT activation. Surprisingly, upon Vav-1 overexpression CD28 ligation sufficed to activate NF-AT in the absence of TCR engagement. This effect was not mediated by overexpression of ZAP-70 nor of SLP 76 but necessitated the intracellular tail of CD28, the intactness of the TCR proximal signaling cascade, the Src-homology domain 2 (SH2) domain of Vav-1, and SLP-76 phosphorylation, an event which was favored by Vav-1 itself. Cells overexpressing Vav-1 formed lamellipodia and microspikes reminiscent of Rac-1 and Cdc42 activation, respectively, for which the SH2 domain of Vav-1 was dispensable. Together, these data suggest that CD28 engagement activates Vav-1 to boost TCR signals through a synergistic cooperation between Vav-1 and SLP-76 and probably via cortical actin changes to facilitate the organization of a signaling zone. PMID- 11034389 TI - Conformational epitopes on the diabetes autoantigen GAD65 identified by peptide phage display and molecular modeling. AB - The major diabetes autoantigen, glutamic acid decarboxylase (GAD65), contains a region of sequence similarity, including six identical residues PEVKEK, to the P2C protein of coxsackie B virus, suggesting that cross-reactivity between coxsackie B virus and GAD65 can initiate autoimmune diabetes. We used the human islet cell mAbs MICA3 and MICA4 to identify the Ab epitopes of GAD65 by screening phage-displayed random peptide libraries. The identified peptide sequences could be mapped to a homology model of the pyridoxal phosphate (PLP) binding domain of GAD65. For MICA3, a surface loop containing the sequence PEVKEK and two adjacent exposed helixes were identified in the PLP binding domain as well as a region of the C terminus of GAD65 that has previously been identified as critical for MICA3 binding. To confirm that the loop containing the PEVKEK sequence contributes to the MICA3 epitope, this loop was deleted by mutagenesis. This reduced binding of MICA3 by 70%. Peptide sequences selected using MICA4 were rich in basic or hydroxyl-containing amino acids, and the surface of the GAD65 PLP-binding domain surrounding Lys358, which is known to be critical for MICA4 binding, was likewise rich in these amino acids. Also, the two phage most reactive with MICA4 encoded the motif VALxG, and the reverse of this sequence, LAV, was located in this same region. Thus, we have defined the MICA3 and MICA4 epitopes on GAD65 using the combination of phage display, molecular modeling, and mutagenesis and have provided compelling evidence for the involvement of the PEVKEK loop in the MICA3 epitope. PMID- 11034390 TI - Structure-guided identification of C3d residues essential for its binding to complement receptor 2 (CD21). AB - A vital role for complement in adaptive humoral immunity is now beyond dispute. The crucial interaction is that between B cell and follicular dendritic cell resident complement receptor 2 (CR2, CD21) and its Ag-associated ligands iC3b and C3dg, where the latter have been deposited as a result of classical pathway activation. Despite the obvious importance of this interaction, the location of a CR2 binding site within C3d, a proteolytic limit fragment of C3dg retaining CR2 binding activity, has not been firmly established. The recently determined x-ray structure of human C3d suggested a candidate site that was remote from the site of covalent attachment to Ag and consisted of an acidic residue-lined depression, which accordingly displays a significant electronegative surface potential. These attributes were consistent with the known ionic strength dependence of the CR2 C3d interaction and with the fact that a significant electropositive surface was apparent in a modeled structure of the C3d-binding domains of CR2. Therefore, we have performed an alanine scan of all of the residues within and immediately adjacent to the acidic pocket in C3d. By testing the mutant iC3b molecules for their ability to bind CR2, we have identified two separate clusters of residues on opposite sides of the acidic pocket, specifically E37/E39 and E160/D163/I164/E166, as being important CR2-contacting residues in C3d. Within the second cluster even single mutations cause near total loss of CR2 binding activity. Consistent with the proposed oppositely charged nature of the interface, we have also found that removal of a positive charge immediately adjacent to the acidic pocket (mutant K162A) results in a 2-fold enhancement in CR2 binding activity. PMID- 11034391 TI - A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5: molecular, immunological, and structural characterization of the epitope-containing domain. AB - Almost 90% of grass pollen-allergic patients are sensitized against group 5 grass pollen allergens. We isolated a monoclonal human IgE Fab out of a combinatorial library prepared from lymphocytes of a grass pollen-allergic patient and studied its interaction with group 5 allergens. The IgE Fab cross-reacted with group 5A isoallergens from several grass and corn species. By allergen gene fragmentation we mapped the binding site of the IgE Fab to a 11.2-kDa N-terminal fragment of the major timothy grass pollen allergen Phl p 5A. The IgE Fab-defined Phl p 5A fragment was expressed in Escherichia coli and purified to homogeneity. Circular dichroism analysis revealed that the rPhl p 5A domain, as well as complete rPhl p 5A, assumed a folded conformation consisting predominantly of an alpha helical secondary structure, and exhibited a remarkable refolding capacity. It reacted with serum IgE from 76% of grass pollen-allergic patients and revealed an extremely high allergenic activity in basophil histamine release as well as skin test experiments. Thus, the rPhl p 5A domain represents an important allergen domain containing several IgE epitopes in a configuration optimal for efficient effector cell activation. We suggest the rPhl p 5A fragment and the corresponding IgE Fab as paradigmatic tools to explore the structural requirements for highly efficient effector cell activation and, perhaps later, for the development of generally applicable allergen-specific therapy strategies. PMID- 11034393 TI - Functional analysis of ligand-binding and signal transduction domains of CD69 and CD23 C-type lectin leukocyte receptors. AB - CD69 and CD23 are leukocyte receptors with distinctive pattern of cell expression and functional features that belong to different C-type lectin receptor subfamilies. To assess the functional equivalence of different domains of these structurally related proteins, a series of CD69/CD23 chimeras exchanging the carbohydrate recognition domain, the neck region, and the transmembrane and cytoplasmic domains were generated. Biochemical analysis revealed the importance of the neck region (Cys68) in the dimerization of CD69. Functional analysis of these chimeras in RBL-2H3 mast cells and Jurkat T cell lines showed the interchangeability of structural domains of both proteins regarding Ca2+ fluxes, serotonin release, and TNF-alpha synthesis. The type of the signal transduced mainly relied on the cytoplasmic domain and was independent of receptor oligomerization. The cytoplasmic domain of CD69 transduced a Ca2+-mediated signaling that was dependent on the extracellular uptake of Ca2+. Furthermore, a significant production of TNF-alpha was induced through the cytoplasmic domain of CD69 in RBL-2H3 cells, which was additive to that promoted via FcepsilonRI, thus suggesting a role for CD69 in the late phase of reactions mediated by mast cells. Our results provide new important data on the functional equivalence of homologous domains of these two leukocyte receptors. PMID- 11034392 TI - Phosphatidylinositol 3-kinase and NF-kappa B/Rel are at the divergence of CD40 mediated proliferation and survival pathways. AB - CD40 receptor ligation evokes several crucial outcomes for the fate of an activated B cell, including proliferation and survival. Although multiple signaling molecules in the CD40 pathways have been identified, their specific roles in regulating proliferation and maintaining cell viability are still obscure. In this report, we demonstrate that the activation of both phosphatidylinositol 3-kinase (PI-3K) and NF-kappaB/Rel transcription factors is crucial for CD40-mediated proliferation. Furthermore, our data indicate that PI 3K is indispensable for CD40-mediated NF-kappaB/Rel activation. This is achieved via activation of AKT and the degradation of IkappaBalpha. Furthermore, we show that PI-3K activity is necessary for the degradation of cyclin-dependent kinase inhibitor p27kip. Therefore, both of these events comprise the mechanism by which PI-3K controls cell proliferation. In contrast to the absolute requirement of PI 3K and NF-kappaB/Rel for proliferation, these signaling molecules are only partially responsible for CD40-mediated survival, as blocking of PI-3K activity did not lead to apoptosis of anti-CD40-treated cells. However, the PI-3K/NF kappaB pathway is still required for CD40-induced Bcl-X gene expression. Taken together, our data indicate that multiple survival pathways are triggered via this receptor, whereas NF-kappaB/Rel and PI-3K are crucial for CD40-induced proliferation. PMID- 11034394 TI - Alanine scanning mutants of rat proinsulin I show functional diversity of anti insulin monoclonal antibodies. AB - In contrast to autoantibodies that are functionally silenced or deleted, IgG Abs that react with autologous insulin routinely follow hormone administration and arise spontaneously in autoimmune (type I) diabetes mellitus. To understand Ab interactions with autologous insulin, rat proinsulin I and 32 alanine substituted analogues were expressed as fusion proteins and used to examine 16 anti-insulin mAb in ELISA. The results identify several amino acid residues that contribute to binding by a large majority (>75%) of mAb, although no single residue is uniformly required for binding by all mAb. Replacements at charged or polar residues on the insulin surface including A4 (Asp), A5 (Gln), A9 (Ser) A12 (Ser), A17 (Gln), A18 (Asn), B13 (Glu), and B21 (Glu) consistently decreased mAb binding. Single alanine substitutions at positions A16 (Leu), A11 (Cys), B8 (Gly), and B15 (Leu) that are predicted to alter the core structure or chain folding vary widely in their impact on Ab binding. mAb that bind insulin preferentially on solid phase (i.e., ELISA) are highly sensitive to replacement of single residues, and substitutions that alter conformation abolish binding. In contrast, high affinity mAb that bind insulin in solution are relatively insensitive to substitutions at single residues, and they maintain binding to all mutants, including those with disrupted conformation. For such high affinity mAb, replacement of long hydrophobic side chains can augment binding, suggesting mAb interactions with insulin include an induced fit. Thus, the ability of insulin to function as a "molten globule" may contribute to the diversity and autoreactivity of the anti-insulin repertoire. PMID- 11034395 TI - Signal joint formation is also impaired in DNA-dependent protein kinase catalytic subunit knockout cells. AB - The effort to elucidate the mechanism of V(D)J recombination has given rise to a dispute as to whether DNA-dependent protein kinase catalytic subunit (DNA-PKcs) contributes to signal joint formation (sjf). Observations reported to date are confusing. Analyses using DNA-PKcs-deficient cells could not conclude the requirement of DNA-PKcs for sjf, because sjf can be formed by end-joining activities which are diverse among cells other than those participating in V(D)J recombination. Here, we observed V(D)J recombination in DNA-PKcs knockout cells and showed that both signal and coding joint formation were clearly impaired in the cells. Subsequently, to directly demonstrate the requirement of DNA-PKcs for sjf, we introduced full-length cDNA of DNA-PKcs into the knockout cells. Furthermore, several mutant DNA-PKcs cDNA constructs designed from mutant cell lines (irs-20, V3, murine scid, and SX9) were also introduced into the cells to obtain further evidence indicating the involvement of DNA-PKcs in sjf. We found as a result that the full-length cDNA complemented the aberrant sjf and that the mutant cDNAs constructs also partially complemented it. Lastly, we looked at whether the kinase activity of DNA-PKcs is necessary for sjf and, as a result, demonstrated a close relationship between them. Our observations clearly indicate that the DNA-PKcs controls not only coding joint formation but also the sjf in V(D)J recombination through its kinase activity. PMID- 11034396 TI - Restriction in V kappa gene use and antigen selection in anti-myeloperoxidase response in mice. AB - Anti-neutrophil cytoplasmic Abs, directed primarily toward myeloperoxidase (MPO) and proteinase 3, are detected in the majority of patients with distinct forms of small vessel vasculitides and pauci-immune necrotizing glomerulonephritis. However, the origin of these autoantibodies remains unknown. We studied the V region gene use in murine anti-MPO Abs derived from Spontaneous Crescentic Glomerulonephritis/Kinjoh mice. A total of 13 anti-MPO-producing hybridomas were generated from four unimmunized mice. Ten of the 13 hybridomas (corresponding to 3 of 4 clones) expressed Vkappa1C but differed in their use of VH genes. The remaining three hybridomas expressed a Vkappa5 gene. Anti-MPO hybridomas from individual mice were derived from single clones as deduced by sequence similarity and splice-site identity. We found a statistically significant bias of amino acid replacement mutations to the complementarity-determining regions (CDR) in the Vkappa1C-expressing hybridomas. Intriguingly, all 10 Vkappa1C hybridomas share a lysine to glutamate mutation in the CDR1. To determine the effects of somatic V gene mutations on binding to MPO, we generated an anti-MPO Ab with an unmutated Vkappa1C L chain and compared its ability to bind MPO with its mutated counterpart. The mutated hybridoma-derived Ab has a 4.75-fold higher avidity for MPO than the unmutated Ab. These results suggest that: 1) the L chain plays a dominant role in determining Ab specificity to MPO, 2) the anti-MPO Ab response is oligoclonal, consistent with Ag selection, and 3) MPO is a driving Ag in the murine anti-MPO Ab response. PMID- 11034397 TI - Mechanism of IL-4-mediated up-regulation of the polymeric Ig receptor: role of STAT6 in cell type-specific delayed transcriptional response. AB - The polymeric IgR (pIgR) mediates transport of dimeric IgA and pentameric IgM across mucosal epithelia, thereby generating secretory Abs. Its expression is up regulated at the transcriptional level by IL-4 in HT-29 cells. In this study, we demonstrate that IL-4 mediates up-regulation of human pIgR through a 554-bp IL-4 responsive enhancer in intron 1. Mutation of a binding site for STAT-6 within this region abolished IL-4-induced enhancement, while an adjacent putative C/EBP site was dispensable. IL-4 treatment induced binding of STAT6 to the intronic STAT6 site, but cooperation with nearby upstream and downstream DNA elements was required for IL-4 responsiveness. Furthermore, IL-4-mediated increased transcription of the pIgR-derived enhancer, like the endogenous pIgR gene, required de novo protein synthesis. Interestingly, a conditionally active form of STAT6 sufficed to activate a pIgR-derived enhancer in HT-29 cells, but not in Cos 1 cells, suggesting a requirement for cell type-specific factors. Thus, STAT6 activation mediates a delayed transcriptional enhancement of pIgR by induction of a de novo synthesized protein that cooperates with STAT6 itself bound to its cognate DNA element in intron 1. This mechanism may represent a general strategy for how pleiotropic cytokines elicit cell type-specific transcriptional responses. PMID- 11034399 TI - Allergy-associated polymorphisms of the Fc epsilon RI beta subunit do not impact its two amplification functions. AB - Two variants of the beta-chain of the high affinity IgE receptor Fc epsilon RI, I181L-V183L and E237G, have been found associated with allergy. We have previously shown that the beta-chain plays at least two distinct amplifier functions. It amplifies Fc epsilon RI surface expression and signaling, resulting in an estimated 12- to 30-fold amplification of downstream events. To test the hypothesis that the I181L-V183L and E237G beta variants may be functionally relevant and could directly contribute to an allergic phenotype, we have evaluated the functional impact of the beta variants on the two amplifier functions of beta. We found that these variants have no direct effect on the beta amplifier functions. However, the possibility remains that these variants are in linkage disequilibrium with other more relevant polymorphisms or are affecting unknown beta-chain functions. PMID- 11034398 TI - Identification of a novel cytokine response element in the human IFN regulatory factor-1 gene promoter. AB - The present study investigates the regulatory mechanisms involved in the cooperation between IFN-gamma and TNF-alpha to promote transcription from IFN regulatory factor-1 (IRF-1). A transient transfection analysis revealed that the region between -218 and -144, where +1 is the transcription start site, as well as previously reported downstream elements, ppkappaB and IFN-gamma activation site/kappaB, were required for the optimal response to the two cytokines. A subsequent DNase I footprint analysis showed that the region between -171 and 144 was inducibly protected with stimulation by TNF-alpha, and this protection was significantly enhanced with the combination of IFN-gamma and TNF-alpha. In an EMSA with the protected region as a probe, a TNF-alpha-inducible complex (C1) and an IFN-gamma-inducible complex (C2), but no synergy-specific DNA-protein complexes, were recognized. The C1 complex consisted of a pre-existing factor (p65/p50), whereas the C2 complex consisted of a newly synthesized IRF-1-related factor. A methylation interference assay revealed the critical G residues (from 167 to -151) for the DNA-protein complex formation specific to the cytokine response, and within this region the novel kappaB sequence, the promoter distal kappaB (pdkappaB) element (5'-GGGGAAG TAC-3'), was identified. Because the base substitutions over the pdkappaB region (from -171 to -144) affected not only the TNF-alpha-response but also that of IFN-gamma, this region might contribute to the cooperative action of the NF-kappaB subunits with the IRF-1-related factor. Finally, we demonstrated that none of the cis-acting elements, ppkappaB, pdkappaB, or IFN-gamma activation site/kappaB, is dispensable for the optimal synergism in response to IFN-gamma and TNF-alpha. PMID- 11034400 TI - Group B Streptococcus induces apoptosis in macrophages. AB - Group B Streptococcus (GBS) is a pathogen that has developed some strategies to resist host immune defenses. Because phagocytic killing is an important pathogenetic mechanism for bacteria, we investigated whether GBS induces apoptosis in murine macrophages. GBS type III strain COH31 r/s (GBS-III) first causes a defect in cell membrane permeability, then at 24 h, apoptosis. Apoptosis was confirmed by several techniques based on morphological changes and DNA fragmentation. Cytochalasin D does not affect apoptosis, suggesting that GBS-III needs not be within the macrophage cytoplasm to promote apoptosis. Inhibition of host protein synthesis prevents apoptosis, whereas inhibition of caspase-1 or -3, does not. Therefore, GBS can trigger an apoptotic pathway independent of caspase 1 and -3, but dependent on protein synthesis. Inhibition of apoptosis by EGTA and PMA, and enhancement of apoptosis by calphostin C and GF109203X suggests that an increase in the cytosolic calcium level and protein kinase C activity status are important in GBS-induced apoptosis. Neither alteration of plasma membrane permeability nor apoptosis were induced by GBS grown in conditions impeding hemolysin expression or when we used dipalmitoylphosphatidylcholine, which inhibited GBS beta-hemolytic activity, suggesting that GBS beta-hemolysin could be involved in apoptosis. beta-Hemolysin, by causing membrane permeability defects, could allow calcium influx, which initiates macrophage apoptosis. GBS also induces apoptosis in human monocytes but not in tumor lines demonstrating the specificity of its activity. This study suggests that induction of macrophage apoptosis by GBS is a novel strategy to overcome host immune defenses. PMID- 11034401 TI - Distinct effects of surfactant protein A or D deficiency during bacterial infection on the lung. AB - Mice lacking surfactant protein (SP)-A (SP-A-/-) or SP-D (SP-D-/-) and wild-type mice were infected with group B streptococcus or Haemophilus influenzae by intratracheal instillation. Although decreased killing of group B streptococcus and H. influenzae was observed in SP-A-/- mice but not in SP-D-/- mice, deficiency of either SP-A or SP-D was associated with increased inflammation and inflammatory cell recruitment in the lung after infection. Deficient uptake of bacteria by alveolar macrophages was observed in both SP-A- and SP-D-deficient mice. Isolated alveolar macrophages from SP-A-/- mice generated significantly less, whereas those from SP-D-/- mice generated significantly greater superoxide and hydrogen peroxide compared with wild-type alveolar macrophages. In SP-D-/- mice, bacterial killing was associated with increased lung inflammation, increased oxidant production, and decreased macrophage phagocytosis. In contrast, in the absence of SP-A, bacterial killing was decreased and associated with increased lung inflammation, decreased oxidant production, and decreased macrophage phagocytosis. Increased oxidant production likely contributes to effective bacterial killing in the lungs of SP-D-/- mice. The collectins, SP-A and SP-D, play distinct roles during bacterial infection of the lung. PMID- 11034402 TI - Role of cystic fibrosis transmembrane conductance regulator in pulmonary clearance of Pseudomonas aeruginosa in vivo. AB - Cystic fibrosis (CF)2 is a fatal genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) that is commonly associated with chronic pulmonary infections with mucoid Pseudomonas aeruginosa (PA). To test the hypothesis that CFTR plays a direct role in PA adhesion and clearance, we have used mouse lines expressing varying levels of human (h) or mouse (m) CFTR. A subacute intratracheal dose of 3 x 10(6) bacteria was cleared with similar kinetics in control wild-type (WT) and transgenic mice overexpressing hCFTR in the lung from the surfactant protein C (SP-C) promoter (SP-C-hCFTR+/-). In a second series of experiments, the clearance of an acute intratracheal dose of 1.5 x 10(7) PA bacteria was also similar in WT, hemizygous SP-C-hCFTR+/-, and bitransgenic gut-corrected FABP-hCFTR+/+-mCFTR-/-, the latter lacking expression of mCFTR in the lung. However, a small but significant decrease in bacterial killing was observed in lungs of homozygote SP-C-hCFTR+/+ mice. Lung pathology in both WT and SP-C-hCFTR+/+ mice was marked by neutrophilic inflammation and bacterial invasion of perivascular and subepithelial compartments. Bacteria were associated primarily with leukocytes and were not associated with alveolar type II or bronchiolar epithelial cells, the cellular sites of SP-C-hCFTR+/+ transgene expression. The results indicate that there is no direct correlation between levels of CFTR expression and bacterial clearance or association of bacteria with epithelial cells in vivo. PMID- 11034403 TI - Exaggerated human monocyte IL-10 concomitant to minimal TNF-alpha induction by heat-shock protein 27 (Hsp27) suggests Hsp27 is primarily an antiinflammatory stimulus. AB - Unlike more well-studied large heat shock proteins (hsp) that induce both T cell antiinflammatory (IL-10, IL-4) and macrophage proinflammatory (TNF-alpha, IL-15, IL-12) cytokines, hsp27, a small hsp, has been primarily identified as a substrate of mitogen-activated protein kinase-activated protein kinase-2 involved in the p38 signaling pathway and activated during monocyte IL-10 production. Hsp27 can also act as an endogenous protein circulating in the serum of breast cancer patients and a protein whose induction correlates to protection from LPS shock. However, the cytokine-stimulating properties of hsp27 have been unexplored. In this study, exogenous hsp27 is demonstrated for the first time as a potent activator of human monocyte IL-10 production, but only a modest inducer of TNF-alpha. Although exogenous hsp27 stimulation activated all three monocyte mitogen-activated protein kinase pathways (extracellular signal-related kinase (ERK) 1/2, c-Jun N-terminal kinase, and p38), only p38 activation was sustained and required for hsp27 induction of monocyte IL-10, while both ERK 1/2 and p38 activation were required for induction of TNF-alpha when using the p38 inhibitor SB203580 or the ERK inhibitor PD98059. Hsp27's transient activation of the c-Jun N-terminal kinase pathway, which can down-regulate IL-10, may contribute to its potent IL-10 induction. Hsp27's ERK 1/2 activation was also less sustained than activation by stimuli like LPS, possibly contributing to its modest TNF-alpha induction. The failure of either PD98059 or anti-TNF-alpha Ab to substantially inhibit IL-10 induction implied that hsp27 induces IL-10 via activation of p38 signaling independently of TNF-alpha activation and may be predominantly an antiinflammatory monokine stimulus. PMID- 11034404 TI - IL-1 beta attenuates IFN-alpha beta-induced antiviral activity and STAT1 activation in the liver: involvement of proteasome-dependent pathway. AB - IFN-alphabeta is the only established treatment for viral hepatitis; however, more than 60% of patients are poorly responsive. Because viral hepatitis is associated with inflammation, we hypothesized that inflammation may attenuate the efficacy of IFN therapy. To test this hypothesis, the effect of IL-1beta, one of the major proinflammatory cytokines, on IFN signaling pathway in the liver was examined. Administration of IL-1beta in vivo attenuated IFN-alphabeta-induced STAT1 tyrosine phosphorylation in the liver but not in the spleen. The inhibitory action of IL-1beta in vivo was not affected by depleting hepatic Kupffer cells, suggesting that IL-1beta may directly target IFN-alphabeta signaling in hepatocytes. Indeed, pretreatment of human hepatocellular carcinoma HepG2 cells with IL-1beta suppressed IFN-alphabeta-induced antiviral activity and antiviral protein MxA mRNA expression. Furthermore, IL-1beta attenuated IFN-alphabeta induced STAT1 binding and tyrosine phosphorylation without affecting the level of STAT1 protein. This inhibitory effect can be reversed by pretreatment with either proteasome inhibitors or transfection of dominant negative NF-kappaB inducing kinase mutants. Taken together, these findings suggest that IL-1beta attenuates IFN-alphabeta-induced STAT1 activation by a proteasome-dependent mechanism. In view of high levels of IL-1beta in the serum or within the liver of patients with chronic liver diseases, attenuation of IFN-alphabeta signaling in the liver by IL 1beta could be one of the mechanisms underlying the resistance to IFN therapy in chronic hepatitis C, and IL-1beta could be a potential therapeutic target for improving the efficacy of IFN therapy. PMID- 11034405 TI - Granzyme activity in the inflamed lung is not controlled by endogenous serine proteinase inhibitors. AB - Numerous lung diseases, such as hypersensitivity pneumonitis (HP), are characterized by the presence of activated alveolar CTL and NK cells. Since these cells produce granzymes, granzyme A and B levels in bronchoalveolar lavage (BAL) fluids from 14 normal subjects and 12 patients with HP were measured by ELISA. Median (range) BAL granzyme A and B levels were 4 (0-37) and 0 (0-6) pg/ml in normal subjects. BAL granzyme levels were significantly higher in HP patients, being at 74 (0-1,889) and 10 (0-78) pg/ml for granzymes A and B, respectively. In vitro, neither of the three main serine protease inhibitors of the lung, namely alpha1-antitrypsin, secretory leukocyte protease inhibitor, and elafin, showed any effect on granzyme A or B activity. In addition, granzyme A was shown to be fully active in BAL fluids. Hence, these data show that granzyme activity may be poorly controlled by protease inhibitors in inflamed tissues. Thus, granzymes could contribute to tissue remodeling and inflammation characterizing HP. PMID- 11034406 TI - Impaired IFN-gamma production in IFN regulatory factor-1 knockout mice during endotoxemia is secondary to a loss of both IL-12 and IL-12 receptor expression. AB - Mice with a targeted mutation in the gene that encodes the transcription factor IFN regulatory factor-1 (IRF-1) were used to assess the contribution of IRF-1 to IL-12-dependent and IL-12-independent pathways of IFN-gamma production. In response to LPS, IRF. 1-/- mice produced less IL-12 p40, IL-12 p35, and IFN-gamma mRNA in the liver than IRF-1+/+ mice. While pulmonary IFN-gamma mRNA levels were also mitigated in IRF-1-/- mice, pulmonary IL-12 p40 and IL-12 p35 mRNA were not dysregulated. Circulating IL-12 p70 and IFN-gamma levels were profoundly attenuated in LPS-challenged IRF-1-/- mice. Further analysis revealed a major deficiency in hepatic IL-12Rbeta1 and IL-12Rbeta2 mRNA expression as well as pulmonary IL-12Rbeta1 mRNA expression in LPS-challenged IRF-1-/- mice. In vitro, IFN-gamma up-regulated IL-12Rbeta1 mRNA in macrophages from IRF-1+/+, but not IRF 1-/-, mice. IFN-gamma-induced IL-12Rbeta2 mRNA expression was also diminished in macrophages from IRF-1-/- mice. In contrast to IRF-1+/+ mice, administration of exogenous IL-12 to IRF-1-/- mice resulted in reduced serum IFN-gamma and hepatic and pulmonary IFN-gamma mRNA, demonstrating that loss of IL-12R results in diminished IL-12 responsiveness. While LPS-challenged IRF-1-/- mice also had reduced IL-15 mRNA levels, serum IL-18 responses were intact. Finally, induction of IRF-1 mRNA by LPS in livers of IFN-gamma knockout mice were markedly attenuated, suggesting a feedback amplification loop. These studies indicate that IRF-1 deficiency disrupts both IL-12-dependent and -independent pathways of IFN gamma production and that IRF-1 is a critical transcription factor involved in the regulation of not only IL-12, but also IL-12R. PMID- 11034407 TI - Nitric oxide-dependent ribosomal RNA cleavage is associated with inhibition of ribosomal peptidyl transferase activity in ANA-1 murine macrophages. AB - NO can regulate specific cellular functions by altering transcriptional programs and protein reactivity. With respect to global cellular processes, NO has also been demonstrated to inhibit total protein synthesis and cell proliferation. The underlying mechanisms are unknown. In a system of ANA-1 murine macrophages, iNOS expression and NO production were induced by exposure to endotoxin (LPS). In selected instances, cells were exposed to an exogenous NO donor, S-nitroso-N acetylpenicillamine or a substrate inhibitor of NO synthesis. Cellular exposure to NO, from both endogenous and exogenous sources, was associated with a significant time-dependent decrease in total protein synthesis and cell proliferation. Gene transcription was unaltered. In parallel with decreased protein synthesis, cells exhibited a distinctive cleavage pattern of 28S and 18S rRNA that were the result of two distinct cuts in both 28S and 18S rRNA. Total levels of intact 28S rRNA, 18S rRNA, and the composite 60S ribosome were significantly decreased in the setting of cell exposure to NO. Finally, 60S ribosome-associated peptidyl transferase activity, a key enzyme for peptide chain elongation, was also significantly decreased. Our data suggest that NO-mediated cleavage of 28S and 18S rRNA results in decreased 60S ribosome associated peptidyl transferase activity and inhibition of total protein synthesis. PMID- 11034408 TI - Cyclooxygenase-2 expression in macrophages: modulation by protein kinase C-alpha. AB - Cyclooxygenase-2 (COX-2) is an inducible enzyme responsible for high levels of PG production during inflammation and immune responses. Previous studies with pharmacological inhibitors suggested a role for protein kinase C (PKC) in PG production possibly by regulating COX-2 expression. In this study, we addressed the role of PKC-alpha in the modulation of COX-2 expression and PGE2 synthesis by the overexpressing of a dominant-negative (DN) mutant of this isoenzyme in the mouse macrophage cell line RAW 264.7. We investigated the effect of various stimuli on COX-2 expression, namely, LPS, IFN-gamma, and the intracellular parasite Leishmania donovani. Whereas LPS-induced COX-2 mRNA and protein expression were down-regulated in DN PKC-alpha-overexpressing clones, IFN-gamma induced COX-2 expression was up-regulated in DN PKC-alpha-overexpressing clones with respect to normal RAW 264.7 cells. Measurements of PGE2 levels revealed a strong correlation between PGE2 secretion and IFN-gamma-induced COX-2 mRNA and protein levels in DN PKC-alpha-overexpressing clones. Taken together, these results suggest a role for PKC-alpha in the modulation of LPS- and IFN-gamma induced COX-2 expression, as well as in IFN-gamma-induced PGE2 secretion. PMID- 11034409 TI - Capsaicin inhibits platelet-activating factor-induced cytosolic Ca2+ rise and superoxide production. AB - Platelet-activating factor (PAF) is an important participant in the inflammatory process. We studied the regulation of PAF activity by capsaicin in human promyelocytic leukemia HL-60 cells. Capsaicin inhibited PAF-induced superoxide production in a concentration-dependent manner. In addition to PAF, the fMLP- and extracellular ATP-induced superoxide productions were inhibited by capsaicin, whereas PMA-induced superoxide production was not affected. In the PAF-stimulated cytosolic Ca2+ increase, capsaicin inhibited in particular the sustained portion of the raised Ca2+ level without attenuation of the peak height. In the absence of extracellular Ca2+, the PAF-induced Ca2+ elevation was not inhibited by capsaicin because capsaicin only inhibited the Ca2+ influx from the extracellular space. In addition, capsaicin did not affect PAF-induced inositol 1,4,5 trisphosphate production, suggesting that phospholipase C activation by PAF is not affected by capsaicin. Store-operated Ca2+ entry (SOCE) induced by thapsigargin was inhibited by capsaicin in a concentration-dependent manner. This capsaicin effect was also observed on thapsigargin-induced Ba2+ and Mn2+ influx. Furthermore, capsaicin's inhibitory effect on the thapsigargin-induced Ca2+ rise overlapped with that of SK&F96365, an inhibitor of SOCE. Both capsaicin and SK&F96365 also inhibited PAF-induced cytosolic superoxide generation in HL-60 cells differentiated by all-trans-retinoic acid. Our data suggest that capsaicin exerts its anti-inflammatory effect by inhibiting SOCE elicited via PLC activation, which occurs upon PAF activation and results in the subsequent superoxide production. PMID- 11034410 TI - Cooperation between decay-accelerating factor and membrane cofactor protein in protecting cells from autologous complement attack. AB - Decay-accelerating factor (DAF or CD55) and membrane cofactor protein (MCP or CD46) function intrinsically in the membranes of self cells to prevent activation of autologous complement on their surfaces. How these two regulatory proteins cooperate on self-cell surfaces to inhibit autologous complement attack is unknown. In this study, a GPI-anchored form of MCP was generated. The ability of this recombinant protein and that of naturally GPI-anchored DAF to incorporate into cell membranes then was exploited to examine the combined functions of DAF and MCP in regulating complement intermediates assembled from purified alternative pathway components on rabbit erythrocytes. Quantitative studies with complement-coated rabbit erythrocyte intermediates constituted with each protein individually or the two proteins together demonstrated that DAF and MCP synergize the actions of each other in preventing C3b deposition on the cell surface. Further analyses showed that MCP's ability to catalyze the factor I-mediated cleavage of cell-bound C3b is inhibited in the presence of factors B and D and is restored when DAF is incorporated into the cells. Thus, the activities of DAF and MCP, when present together, are greater than the sum of the two proteins individually, and DAF is required for MCP to catalyze the cleavage of cell-bound C3b in the presence of excess factors B and D. These data are relevant to xenotransplantation, pharmacological inhibition of complement in inflammatory diseases, and evasion of tumor cells from humoral immune responses. PMID- 11034411 TI - Redundant and segregated functions of granule-associated heparin-binding group II subfamily of secretory phospholipases A2 in the regulation of degranulation and prostaglandin D2 synthesis in mast cells. AB - We herein demonstrate that mast cells express all known members of the group II subfamily of secretory phospholipase A2 (sPLA2) isozymes, and those having heparin affinity markedly enhance the exocytotic response. Rat mastocytoma RBL 2H3 cells transfected with heparin-binding (sPLA2-IIA, -V, and -IID), but not heparin-nonbinding (sPLA2-IIC), enzymes released more granule-associated markers (beta-hexosaminidase and histamine) than mock- or cytosolic PLA2alpha (cPLA2alpha)-transfected cells after stimulation with IgE and Ag. Site-directed mutagenesis of sPLA2-IIA and -V revealed that both the catalytic and heparin binding domains are essential for this function. Confocal laser and electron microscopic analyses revealed that sPLA2-IIA, which was stored in secretory granules in unstimulated cells, accumulated on the membranous sites where fusion between the plasma membrane and granule membranes occurred in activated cells. These results suggest that the heparin-binding sPLA2s bind to the perigranular membranes through their heparin-binding domain, and lysophospholipids produced in situ by their enzymatic action may facilitate the ongoing membrane fusion. In contrast to the redundant role of sPLA2-IIA, -IID, and -V in the regulation of degranulation, only sPLA2-V had the ability to markedly augment IgE/Ag-stimulated immediate PGD2 production, which reached a level comparable to that elicited by cPLA2alpha. The latter observation reveals an unexplored functional segregation among the three related isozymes expressed in the same cell population. PMID- 11034412 TI - Macrophage inflammatory protein-2 and KC induce chemokine production by mouse astrocytes. AB - Astrocytes are specialized cells of the CNS that are implicated in the pathogenesis of multiple sclerosis and experimental allergic encephalomyelitis. In acute and relapsing-remitting experimental allergic encephalomyelitis, the neutrophil chemoattractant CXC chemokines macrophage-inflammatory protein (MIP)-2 and KC are associated with reactive astrocytes in the parenchyma. In vitro treatment of primary astrocyte cultures with nanomolar concentrations of MIP-2 or KC markedly up-regulated expression of the monocyte/T cell chemoattractants monocyte chemoattractant protein-1, inflammatory protein-10, and RANTES by a mechanism that includes stabilization of mRNA. Production of TNF-alpha and IL-6 transcripts were also noted, as was autocrine induction of MIP-2 and KC message. In addition, low levels of MIP-1alpha and MIP-1beta were induced following treatment with MIP-2 or KC. These effects are specific to astrocytes as MIP-2 treatment of microglial cells failed to elicit chemokine production. The astrocyte chemokine receptor for MIP-2 has 2.5 nM affinity for ligand. Astrocytes from CXCR2-deficient mice still respond to KC and MIP-2, indicating the presence of an alternative or novel high affinity receptor for these ligands. We propose that this KC/MIP-2 chemokine cascade may contribute to the persistence of mononuclear cell infiltration in demyelinating autoimmune diseases. PMID- 11034413 TI - Studies on a mechanism by which cytosolic phospholipase A2 regulates the expression and function of type IIA secretory phospholipase A2. AB - Although it has been proposed that arachidonate release by several secretory phospholipase A2 (sPLA2) isozymes is modulated by cytosolic PLA2 (cPLA2), the cellular component(s) that intermediates between these two signaling PLA2s remains unknown. Here we provide evidence that 12- or 15-lipoxygenase (12/15 LOX), which lies downstream of cPLA2, plays a pivotal role in cytokine-induced gene expression and function of sPLA2-IIA. The sPLA2-IIA expression and associated PGE2 generation induced by cytokines in rat fibroblastic 3Y1 cells were markedly attenuated by antioxidants that possess 12/15-LOX inhibitory activity. 3Y1 cells expressed 12/15-LOX endogenously, and forcible overexpression of 12/15-LOX in these cells greatly enhanced cytokine-induced expression of sPLA2 IIA, with a concomitant increase in delayed PG generation. Moreover, studies using 293 cells stably transfected with sPLA2-IIA revealed that stimulus dependent hydrolysis of membrane phospholipids by sPLA2-IIA was enhanced by overexpression of 12/15-LOX. These results indicate that the product(s) generated by the cPLA2-12/15-LOX pathway following cell activation may play two roles: enhancement of sPLA2-IIA gene expression and membrane sensitization that leads to accelerated sPLA2-IIA-mediated hydrolysis. PMID- 11034414 TI - GM-CSF regulates bleomycin-induced pulmonary fibrosis via a prostaglandin dependent mechanism. AB - To characterize the role of GM-CSF in pulmonary fibrosis, we have studied bleomycin-induced fibrosis in wild-type mice vs mice with a targeted deletion of the GM-CSF gene (GM-CSF-/- mice). Without GM-CSF, pulmonary fibrosis was worse both histologically and quantitatively. These changes were not related to enhanced recruitment of inflammatory cells because wild-type and GM-CSF-/- mice recruited equivalent numbers of cells to the lung following bleomycin. Interestingly, recruitment of eosinophils was absent in GM-CSF-/- mice. We investigated whether the enhanced fibrotic response in GM-CSF-/- animals was due to a deficiency in an endogenous down-regulator of fibrogenesis. Analysis of whole lung homogenates from saline- or bleomycin-treated mice revealed that GM CSF-/- animals had reduced levels of PGE2. Additionally, alveolar macrophages were harvested from wild-type and GM-CSF-/- mice that had been exposed to bleomycin. Although bleomycin treatment impaired the ability of alveolar macrophages from wild-type mice to synthesize PGE2, alveolar macrophages from GM CSF-/- mice exhibited a significantly greater defect in PGE2 synthesis than did wild-type cells. Exogenous addition of GM-CSF to alveolar macrophages reversed the PGE2 synthesis defect in vitro. Administration of the PG synthesis inhibitor, indomethacin, to wild-type mice during the fibrogenic phase postbleomycin worsened the severity of fibrosis, implying a causal role for PGE2 deficiency in the evolution of the fibrotic lesion. These data demonstrate that GM-CSF deficiency results in enhanced fibrogenesis in bleomycin-induced pulmonary fibrosis and indicate that one mechanism for this effect is impaired production of the potent antifibrotic eicosanoid, PGE2. PMID- 11034415 TI - Eosinophilopoiesis in a murine model of allergic airway eosinophilia: involvement of bone marrow IL-5 and IL-5 receptor alpha. AB - The airway inflammation in asthma is dominated by eosinophils. The aim of this study was to elucidate the contribution of newly produced eosinophils in airway allergic inflammation and to determine mechanisms of any enhanced eosinophilopoiesis. OVA-sensitized BALB/c mice were repeatedly exposed to allergen via airway route. Newly produced cells were identified using a thymidine analog, 5-bromo-2'-deoxyuridine, which is incorporated into DNA during mitosis. Identification of IL-5-producing cells in the bone marrow was performed using FACS. Bone marrow CD3+ cells were enriched to evaluate IL-5-protein release in vitro. Anti-IL-5-treatment (TRFK-5) was given either systemically or directly to the airways. IL-5R-bearing cells were localized by immunocytochemistry. Repeated airway allergen exposure caused prominent airway eosinophilia after three to five exposures, and increased the number of immature eosinophils in the bone marrow. Up to 78% of bronchoalveolar lavage (BAL) granulocytes were 5-bromo-2' deoxyuridine positive. After three allergen exposures, both CD3+ and non-CD3 cells acquired from the bone marrow expressed and released IL-5-protein. Anti-IL 5 given i.p. inhibited both bone marrow and airway eosinophilia. Intranasal administration of anti-IL-5 also reduced BAL eosinophilia, partly via local effects in the airways. Bone marrow cells, but not BAL eosinophils, displayed stainable amounts of the IL-5R alpha-chain. We conclude that the bone marrow is activated by airway allergen exposure, and that newly produced eosinophils contribute to a substantial degree to the airway eosinophilia induced by allergen. Airway allergen exposure increases the number of cells expressing IL-5 protein in the bone marrow. The bone marrow, as well as the lung, are possible targets for anti-IL-5-treatment. PMID- 11034416 TI - Endogenous and exogenous IL-6 inhibit aeroallergen-induced Th2 inflammation. AB - Chronic Th2-dominated inflammation and exaggerated IL-6 production are characteristic features of the asthmatic airway. To understand the processes that are responsible for the chronicity of this response and the role(s) of IL-6 in the regulation of airway Th2 inflammation, we compared the responses induced by OVA in sensitized wild-type mice, IL-6 deficient (-/-) mice, and transgenic mice in which IL-6 was overexpressed in the airway (CC10-IL-6 mice). When compared with wild-type mice, IL-6-/- mice manifest exaggerated inflammation and eosinophilia, increased levels of IL-4, IL-5, and IL-13 protein and mRNA, exaggerated levels of eotaxin, JE/monocyte chemoattractant protein-1, macrophage inflammatory protein-1alpha and -2, and mRNA, increased bronchoalveolar lavage (BAL) TGF-beta1, and exaggerated airway responses to aerosolized methacholine. In contrast, CC10-IL-6 mice, on both C57BL/6 and BALB/c backgrounds, manifest diminished inflammation and eosinophilia, decreased levels of IL-4, IL-5, and IL 13 protein and mRNA, and decreased levels of bronchoalveolar lavage TGF-beta1. IL 6 also decreased the expression of endothelial VCAM-1 and airway responsiveness to methacholine in these animals. These alterations in the IL-6-/- and CC10-IL-6 mice were not associated with significant decreases or increases in the levels of IFN-gamma, respectively. These studies demonstrate that endogenous and exogenous IL-6 inhibit aeroallergen-induced Th2 inflammation and that this inhibition is not mediated by regulatory effects of IFN-gamma. IL-6 may be an important anti inflammatory, counterregulatory, and healing cytokine in the airway. PMID- 11034417 TI - Experimentally induced recruitment of plasmacytoid (CD123high) dendritic cells in human nasal allergy. AB - Recent evidence suggests that the previously enigmatic cell type designated plasmacytoid monocytes can function as dendritic cells and contribute substantially to both innate and adaptive immunity. This cell type has previously been described only in bone marrow, blood, and organized lymphoid tissue, but not at effector sites with direct Ag exposure such as the mucosae. Plasmacytoid dendritic cells (P-DCs) matured in vitro can induce T cells to produce allergy promoting Th2 cytokines; therefore, their possible occurrence in nasal mucosa during experimentally elicited allergic rhinitis was examined. Patients with silent nasal allergy were challenged topically with relevant allergen daily for 7 days. Biopsy specimens as well as blood samples were obtained before and during such provocation, and P-DCs were identified by their high expression of CD123 (IL 3R alpha-chain), together with CD45RA. Our results showed that P-DCs were present in low and variable numbers in normal nasal mucosa but increased dramatically during the allergic reaction. This accumulation concurred with the expression of the L-selectin ligand peripheral lymph node addressin on the mucosal vascular endothelium. The latter observation was particularly interesting in view of the high levels of L-selectin on circulating P-DC precursors and of previous reports suggesting that these cells can enter organized lymphoid tissue via high endothelial venules (which express peripheral lymph node addressin constitutively). Together, our findings suggested that P-DCs are involved in the triggering of airway allergy and that they are directed to allergic lesions by adhesion molecules that normally mediate leukocyte extravasation in organized lymphoid tissue. PMID- 11034418 TI - Eosinophils maintain their capacity to signal and release eosinophil cationic protein upon repetitive stimulation with the same agonist. AB - Eosinophils contain in their granules eosinophil cationic protein (ECP) and other basic proteins that have been implicated in immunity to parasites and pathophysiology of chronic allergic responses. In a model of eosinophil degranulation, we show that eosinophils release ECP upon short-term GM-CSF priming and stimulation with either platelet-activating factor (PAF) or the anaphylatoxin C5a, but not eotaxin. Restimulation with the same agonist (PAF or C5a) was unsuccessful as assessed by monitoring intracellular calcium concentration and ECP release. In contrast, upon an intermediate washing step, eosinophils rapidly transduced PAF and C5a signals followed by significant ECP releases. Ligand-binding studies demonstrated that only a proportion of PAF receptors is internalized upon cell stimulation and that washing of the cells removes the agonist from the cell surface. Upon repetitive stimulation, eosinophils with less than 50% of the original ECP content were obtained. Such eosinophils did not increase cellular ECP levels even in the presence of the eosinophil survival factor GM-CSF in overnight cultures. In vivo studies revealed that eosinophils always express detectable amounts of ECP under chronic inflammatory conditions. In conclusion, we have shown that eosinophils maintain their capacity to degranulate upon repetitive stimulation with the same agonist as long as the receptor is not occupied from a previous stimulation. The cellular content of ECP appears to be a no limiting factor in the case of repetitive stimulation, implying that mature eosinophils may not require a significant ECP resynthesis. PMID- 11034419 TI - Overexpression of CD1d by keratinocytes in psoriasis and CD1d-dependent IFN-gamma production by NK-T cells. AB - The MHC class I-like protein CD1d is a nonpolymorphic molecule that plays a central role in development and activation of a subset of T cells that coexpress receptors used by NK cells (NK-T cells). Recently, T cells bearing NK receptors were identified in acute and chronic lesions of psoriasis. To determine whether NK-T cells could interact with epidermal cells, we examined the pattern of expression of CD1d in normal skin, psoriasis, and related skin disorders, using a panel of CD1d-specific mAbs. CD1d was expressed by keratinocytes in normal skin, although expression was at a relatively low level and was generally confined to upper level keratinocytes immediately beneath the lipid-rich stratum corneum. In contrast, there was overexpression of CD1d in chronic, active psoriatic plaques. CD1d could be rapidly induced on keratinocytes in normal skin by physical trauma that disrupted barrier function or by application of a potent contact-sensitizing agent. Keratinocytes displayed enhanced CD1d following exposure to IFN-gamma. Combining CD1d-positive keratinocytes with human NK-T cell clones resulted in clustering of NK-T cells, and while no significant proliferation ensued, NK-T cells became activated to produce large amounts of IFN-gamma. We conclude that CD1d can be expressed in a functionally active form by keratinocytes and is up regulated in psoriasis and other inflammatory dermatoses. The ability of IFN gamma to enhance keratinocyte CD1d expression and the subsequent ability of CD1d positive keratinocytes to activate NK-T cells to produce IFN-gamma, could provide a mechanism that contributes to the pathogenesis of psoriasis and other skin disorders. PMID- 11034421 TI - CD4+ T lymphocytes with constitutive CD40 ligand in preautoimmune (NZB x NZW)F1 lupus-prone mice: phenotype and possible role in autoreactivity. AB - Lupus disease is marked by B lymphocyte hyperactivity and the production of Abs to dsDNA. The production of these anti-dsDNA Abs is T lymphocyte dependent. However, it is not clear how CD4+ T lymphocytes provide help for B lymphocytes to produce IgG anti-dsDNA Abs. One possible mechanism is suggested by studies showing that human patients with systemic lupus erythematosus and lupus mice have increased numbers of CD40 ligand (CD40L)+ T and B lymphocytes. The results described in this study reveal that young, clinically healthy lupus-prone New Zealand Black x New Zealand White F1 (BWF1) mice have naive CD4+ T cells with preformed CD40L. These cells contribute to a brisk response to immunization and to the production of anti-dsDNA Abs. In vitro experiments revealed that CD4+ T cells with preformed CD40L could, upon stimulation, provide antiapoptotic signals for B cells but could not induce proliferation or reduce activation threshold. These results suggest that the direct target cells for the effect of T cells with preformed CD40L in lupus may not be B lymphocytes. PMID- 11034420 TI - Peptide dose, MHC affinity, and target self-antigen expression are critical for effective immunotherapy of nonobese diabetic mouse prediabetes. AB - Cross-reactive T cells that recognize both Tep69 (dominant nonobese diabetic (NOD) T cell epitope in ICA69 (islet cell autoantigen of 69 kDa)) and ABBOS (dominant NOD T cell epitope in BSA) are routinely generated during human and NOD mouse prediabetes. Here we analyzed how systemic administration of these mimicry peptides affects progressive autoimmunity in adoptively transferred and cyclophosphamide-accelerated NOD mouse diabetes. These models were chosen to approximate mid to late stage prediabetes, the typical status of probands in human intervention trials. Unexpectedly, high dose (100 microg) i.v. ABBOS prevented, while Tep69 exacerbated, disease in both study models. Peptide effects required cognate recognition of endogenous self-Ag, because both treatments were ineffective in ICA69null NOD congenic mice adoptively transferred with wild-type, diabetic splenocytes. The affinity of ABBOS for NOD I-A(g7) was orders of magnitude higher than that of Tep69. This explained 1) the expansion of the mimicry T cell pool following i.v. Tep69, 2) the long-term unresponsiveness of these cells after i.v. ABBOS, and 3) precipitation of the disease after low dose i.v. ABBOS. Disease precipitation and prevention in mid to late stage prediabetes are thus governed by affinity profiles and doses of therapeutic peptides. ABBOS or ABBOS analogues with even higher MHC affinity may be candidates for experimental intervention strategies in human prediabetes, but the dose translation from NOD mice to humans requires caution. PMID- 11034422 TI - B lymphoblastoid cell lines as efficient APC to elicit CD8+ T cell responses against a cytomegalovirus antigen. AB - Potent and readily accessible APC are critical for development of immunotherapy protocols to treat viral disease and cancer. We have shown that B lymphoblastoid cell lines (BLCL) that stably express CMV phosphoprotein 65 (BLCLpp65), as a result of retroviral transduction, can be used to generate ex vivo CTL cultures that possess cytotoxicity against CMV and EBV. In this report, we demonstrate that the EBV-specific cytotoxicity in the BLCLpp65-primed culture had a spectrum of EBV-Ag recognition similar to that of the BLCL-primed counterpart, suggesting that retroviral transduction and expression of the CMV Ag would not compromise the Ag-presenting capacity of BLCL. In addition, BLCLpp65 appeared to present multiple natural pp65 epitopes, because pp65-specific CTL, which recognized different CMV clinical isolates, were generated in BLCLpp65-primed cultures from individuals with various HLA backgrounds. Consistent with a polyclonal expansion of virus-specific CTL, T cell lines established from the BLCLpp65-primed CTL cultures expressed different TCR-Vbeta Although most of the virus-specific T cell isolates were CD8+, EBV-specific CD4+ lines were also established from BLCLpp65 primed cultures. Western blot analysis revealed that the CD8+ lines, but not the CD4+ line, expressed granzyme B, consistent with features of classic CTL. Thus, our results suggested that BLCL stably expressing a foreign Ag might be used as a practical APC to elicit CD8+ T cell responses. PMID- 11034423 TI - Expression of a human coxsackie/adenovirus receptor transgene permits adenovirus infection of primary lymphocytes. AB - Replication-defective adenoviruses are effective vehicles for gene transfer, both for the repair of defective genes and for studies of gene function in primary cells. Many cell types, including lymphocytes, are refractory to adenovirus infection because they lack the Coxsackie/adenovirus receptor (CAR) needed for virus attachment. To extend the advantages of adenovirus-mediated gene transfer to primary lymphoid populations and other cell types lacking endogenous CAR, we produced a mouse that expresses human (h) CAR as a transgene under control of a murine MHC class I promoter. hCAR protein is expressed on T and B lymphocytes from a variety of organs (spleen, lymph node, bone marrow, thymus, and peritoneum). These lymphocytes are susceptible to adenovirus infection, as demonstrated by reporter green fluorescent protein gene expression, with the fraction of expressing cells as high as 70%. Some lymphocyte subpopulations required stimulation subsequent to adenovirus infection for reporter expression. This activation requirement is a restriction imposed by the promoter used in the adenovirus construct. In subpopulations requiring activation, the elongation factor 1 promoter was far superior to a hCMV promoter for directing green fluorescent protein production. We also find that hCAR mRNA is produced in nonlymphoid tissues from all founder lines, including tissues that do not express endogenous murine CAR, suggesting the opportunity for effecting gene delivery to and testing gene function in a wide variety of primary cell types previously resistant to gene transfer. PMID- 11034424 TI - Impairment of STAT activation by IL-12 in a patient with atypical mycobacterial and staphylococcal infections. AB - IL-12 plays a pivotal role in the stimulation of immune responses against intracellular infections. This role is manifested in the increased susceptibility to atypical mycobacterial and salmonella infections among individuals whose lymphocytes lack expression of IL-12Rbeta1. Here, we report on a patient with Mycobacterium avium infection, recurrent Staphylococcus aureus sinusitis, and multiple adverse drug reactions whose T cells were unable to produce IFN-gamma or proliferate in response to IL-12 despite the expression of wild-type IL-12Rbeta1 and IL-12Rbeta2. The defect in these functional responses to IL-12 was selective, as cytolytic activity induced by IL-12 was intact, and lymphocytes were responsive to stimulation by IL-2. An examination of cytokine signaling revealed that STAT4 and extracellular regulated kinase 1 (ERK1) activation by IL-12 was intact, whereas the activation of STAT1, -3, and -5 by IL-12 was lost. This impairment of STAT activation was specific for IL-12, as STAT activation by IL-2, IL-15, and IFN-gamma was unaffected. These findings demonstrate that the activation of STAT4 alone is not sufficient for IL-12-induced IFN-gamma production and proliferation and suggest that other STATs play a role in these responses to IL-12. While the etiology of the impaired IL-12 signaling in this patient has not yet been elucidated, the absence of mutations in IL-12Rbeta1 or IL-12Rbeta2 and the preservation of STAT4 activation raise the possibility that there may be a mutation in an as yet undiscovered component of the IL-12 signaling complex that is normally required for the recruitment and activation of STAT1, -3, and -5. PMID- 11034425 TI - Spontaneous production of C-C chemokines by individuals infected with human T lymphotropic virus type II (HTLV-II) alone and HTLV-II/HIV-1 coinfected individuals. AB - To investigate the immunological features of human T lymphotropic virus type II (HTLV-II) infection and specific mechanisms whereby HTLV-II might influence the progression of HIV-1 disease in coinfected individuals, we have analyzed the production of the C-C chemokines RANTES and macrophage inflammatory proteins 1alpha and 1alpha (MIP-1alpha and MIP-1beta) by PBMCs from HTLV-II-infected and HTLV-II/HIV-1-coinfected individuals. We observed spontaneous production of significant levels of MIP-1alpha and -1beta and, to a lesser extent, RANTES, from individuals infected with HTLV-II alone or with concomitant HIV-1 infection. Spontaneous C-C chemokine production was not observed in PBMCs from uninfected or HIV-1-infected individuals. Although HTLV-II is known to preferentially infect CD8+ lymphocytes in vivo, we observed that whereas RANTES was produced exclusively by the CD8+-enriched fraction, MIP-1alpha and -1beta were produced by both the CD8+-enriched and CD8+-depleted fractions of HTLV-II-infected PBMCs. RT PCR demonstrated active expression of the HTLV-II regulatory protein Tax in the infected CD8+ T lymphocyte population, and it was further shown that Tax transactivates the promoters of MIP-1beta and RANTES. Therefore, it appears that HTLV-II stimulates the production of C-C chemokines both directly at a transcriptional level via the viral transactivator Tax and also indirectly. Although the HTLV-II-infected individuals in this study are all virtually asymptomatic, they certainly display an abnormal immune phenotype. Moreover, our findings suggest that HTLV-II, via chemokine production, would be expected to alter the progression of HIV-1 infection in coinfected individuals. PMID- 11034426 TI - Generation of tumor-reactive CTL against the tumor-associated antigen HER2 using retrovirally transduced dendritic cells derived from CD34+ hemopoietic progenitor cells. AB - Ag-specific CD8+ CTL are crucial for effective tumor rejection. Attempts to treat human malignancies by adoptive transfer of tumor-reactive CTL have been limited due to the difficulty of generating and expanding autologous CTL with defined Ag specificity. The current study examined whether human CTL can be generated against the tumor-associated Ag HER2 using autologous dendritic cells (DC) that had been genetically engineered to express HER2. DC progenitors were expanded by culturing CD34+ hemopoietic progenitor cells in the presence of the designer cytokine HyperIL-6. Proliferating precursor cells were infected by a retroviral vector encoding the HER2 Ag and further differentiated into CD83+ DC expressing high levels of MHC, adhesion, and costimulatory molecules. Retroviral transduction of DC resulted in the expression of the HER2 molecule with a transduction efficiency of 15%. HER2-transduced DC correctly processed and presented the Ag, because HLA-A*0201-positive DC served as targets for CTL recognizing the HLA-A*0201-binding immunodominant peptide HER2(369-377). HER2 transduced DC were used as professional APCs for stimulating autologous T lymphocytes. Following repetitive stimulation, a HER2-specific, HLA-A*0201 restricted CTL line was generated that was capable of lysing HLA-A*0201-matched tumor cells overexpressing HER2. A CD8+ T cell clone could be generated that displayed the same specificity pattern as the parenteral CTL line. The ability to generate and expand HER2-specific, MHC class I-restricted CTL clones using HER2 transduced autologous DC in vitro facilitates the development of adoptive T cell transfer for patients with HER2-overexpressing tumors without the requirement of defining immunogenic peptides. PMID- 11034427 TI - Maximal oxygen uptake and cardiorespiratory response to maximal 400-m free swimming, running and cycling tests in competitive swimmers. AB - BACKGROUND: This study compared the cardiorespiratory response of trained swimmers to 400-m unimpeded front crawl swimming (SW), treadmill running (TR) and ergometer cycling (EC) maximal exercise tests, and evaluated the validity and specificity of a method to measure maximal aerobic power in swimming. METHODS: Two series of experiments were conducted. In series A (n=15), comparisons were made between VO2peak and other cardiorespiratory variables in three maximal tests: after 400-m SW, and during incremental TR and EC. In series B, VO2 peak and related variables were measured after SW and during EC (n=33). RESULTS: No significant differences were observed between VO2peak and VE in the three modes of exercise, although SW values tended to be higher. After SW, maximal ventilatory response was characterized by higher tidal volumes (VT) and lower respiratory rates (fR) as compared with TR and EC. The highest heart rate values (fH) were also observed in TR, followed by EC and SW. In series B, no significant differences were observed either in peak VO2 or VE, but fH was also lower in SW. CONCLUSIONS: A maximal 400-m unimpeded freestyle SW test yields essentially equal or nonsignificantly higher peak VO2 and VE values than during maximal TR or EC tests in trained swimmers. The specific maximal cardiorespiratory response to the SW test is characterized by higher VT, lower fR, and lower fH. Breath-by-breath measurements during the immediate recovery after a 400-m voluntary maximal swim is proposed as a valid and specific test for directly measuring maximal metabolic parameters and evaluating specific maximal aerobic power in swimming. PMID- 11034428 TI - Time limit and time at VO2max' during a continuous and an intermittent run. AB - BACKGROUND: The purpose of this study was to verify, by track field tests, whether sub-elite runners (n=15) could (i) reach their VO2max while running at v50%delta, i.e. midway between the speed associated with lactate threshold (vLAT) and that associated with maximal aerobic power (vVO2max), and (ii) if an intermittent exercise provokes a maximal and/or supra maximal oxygen consumption longer than a continuous one. METHODS: Within three days, subjects underwent a multistage incremental test during which their vVO2max and vLAT were determined; they then performed two additional testing sessions, where continuous and intermittent running exercises at v50%delta were performed up to exhaustion. Subject's gas exchange and heart rate were continuously recorded by means of a telemetric apparatus. Blood samples were taken from fingertip and analysed for blood lactate concentration. RESULTS: In the continuous and the intermittent tests peak VO2 exceeded VO2max values, as determined during the incremental test. However in the intermittent exercise, peak VO2, time to exhaustion and time at VO2max reached significantly higher values, while blood lactate accumulation showed significantly lower values than in the continuous one. CONCLUSIONS: The v50%delta is sufficient to stimulate VO2max in both intermittent and continuous running. The intermittent exercise results better than the continuous one in increasing maximal aerobic power, allowing longer time at VO2max and obtaining higher peak VO2 with lower lactate accumulation. PMID- 11034429 TI - Physiological profile and predictors of cycling performance in ultra-endurance triathletes. AB - BACKGROUND: To report physiological profiles, and investigate the relationship between selected physiological variables and cycling performance in ultra endurance triathletes. METHODS: PARTICIPANTS: ten male (mean+/-SD, age; 32+/-5 years) ultra-endurance triathletes participated in the study. Physiological profiles were compared with 10 male age-matched control subjects. MEASURES: left ventricular structure (wall thickness [LVPWd], internal diameter [LVIDd], and mass [LVM]) and function (diastolic filling, fractional shortening, and stroke volume [SV]) were assessed using standard M-Mode, 2D, and Doppler echocardiography. Maximal and sub-maximal exercise gas exchange responses were measured on-line during a maximal ramping cycle-ergometer exercise test. RESULTS: Ultra-endurance triathletes demonstrated significantly larger LVPWd, LVIDd, LVM, SV, VO2max anaerobic threshold (AT), and power to body-mass ratio compared with controls. Cycling performance for both Ironman and half Ironman were significantly correlated with LVPWd, LVM, and SV. LVIDd was significantly correlated Ironman cycle time alone. Oxygen consumption (VO2) at AT, percentage of VO2max at AT, and peak power to bodymass ratio were significantly correlated to bike finish time in the half Ironman, but not Ironman. CONCLUSIONS: The correlation between cycling performance, LVM and SV suggests that the more conditioned athletes may be better able to maintain a high cardiac output during prolonged cycling. Sub-maximal gas exchange responses are predictors of cycling performance for the half-Ironman but not the Ironman. These results suggest that other factors including the longer duration swim prior to the cycling component, may impact upon cycle performance. PMID- 11034430 TI - Cross transfer effects of muscular endurance during training and detraining. AB - BACKGROUND: To clarify 1) how the cross-transfer effect, obtained in a contralateral untrained forearm through a 4-week ipsilateral endurance training regimen, is changed during detraining; and 2) how blood flow to the untrained limb is related to the transfer effect of muscular endurance during training and detraining periods. METHODS: Training regimen: hand-grip training by means of a hand-ergometer with a work-load of 1/3 of the maximum handgrip strength 5 times a week for 4 weeks. Blood flow: a mercury-in-rubber strain-gauge for venous occlusion plethysmography. MEASURES: 1) maximal number of contractions to determine the muscular endurance; 2) reactive hyperaemic blood flow response (RHBF3) to determine whether maximal vasodilatory capacity would be changed in both the forearms post-training and detraining; and 3) maximal work-related blood flow. RESULTS: We found significant increments both in the muscle endurance and the maximal work-related blood flow not only in the trained (+125%, +30%) but also in the untrained (+40%, +19%) forearms. During detraining, we found decreases both in the muscle endurance and the maximal work-related blood flow ( 22%, p<0.01; -16%, p=0.053) of the trained forearm. However, in the untrained arm (-3%, NS) the cross-transfer effect of muscular endurance remained unchanged despite a drop in the maximal work-related blood flow (-17%, p<0.05). The RHBF3 did not change in either of the forearms during the whole periods. CONCLUSIONS: These findings suggest that the maintenance of the cross transfer effect of muscle endurance during detraining cannot be explained on the basis of changes in forearm blood flow. PMID- 11034431 TI - Metabolic cost of free weight circuit weight training. AB - BACKGROUND: Free weight circuit weight training (CWT) classes are popular group exercise classes designed to improve aerobic capacity, body composition, and muscular strength and endurance. The purpose of this investigation was to determine the training intensity and caloric expenditure associated with free weight CWT. METHODS: Twelve males and eighteen females (age 25.1+/-6.6 years) participated in a Bruce treadmill test to measure VO2max (47.9+/-10.6 ml/kg min). Subjects subsequently performed a learning trial, exercising to a 14 minute video taped free weight CWT sequence which included squats and upper body exercises performed consecutively. All subjects then completed two randomly assigned video exercise bouts with light resistance (LR = 1.4 kg for males and females) and moderate resistance (MR = 5.9 kg for females and 10.5 kg for males), loads recommended by instructors for sedentary and fit individuals, respectively. RESULTS: Statistical analysis by RM ANOVA (p<0.0036) revealed significant increases in absolute and relative VO2, HR, and energy expenditure at MR as compared to LR for males and females. Mean VO2 and caloric expenditure values at MR were 15.7+/-2.3 ml/kg min and 6.21+/-1.01 kcal/min for males and 13.5+/-1.4 ml/kg min and 4.04+/-1.45 kcal/min for females. Associated HR responses were 129.5+/-18.4 and 119.2+/-19.4 bpm for males and females, respectively. The training stimulus was <32% VO2max, significantly below ACSM recommendations (50% VO2max) for improving cardiovascular fitness; HR criteria (60% HRmax), however were met. Free weight CWT performed with loads < or =10.5 kg may not provide a sufficient cardiovascular training stimulus. CONCLUSIONS: HR should not be used to assess exercise intensity in these classes. PMID- 11034432 TI - Effects of specific warm-up at various intensities on energy metabolism during subsequent exercise. AB - BACKGROUND: To investigate the effects of specific warm-up at various intensities on energy metabolism during subsequent intense exercise. METHODS: EXPERIMENTAL DESIGN: specific warm-up was consisted of 3 sets of wrist flexions for 5 min, with each set followed by a 3-min rest. The intensity of specific warm-up was set at 20%, 30% or 40% of maximal voluntary contraction (MVC). The subjects then performed a set of wrist flexions at 60% MVC for 4 min as the criterion exercise. For the control experiment, criterion exercise was done without specific warm-up. PARTICIPANTS: Five healthy volunteers. MEASUREMENTS: using phosphorus-31 magnetic resonance spectroscopy, spectra were obtained from the wrist flexor muscles to determine the ratio of inorganic phosphate to phosphocreatine (Pi/PCr) and intracellular pH. RESULTS: The Pi/PCr during the criterion exercise after specific warm-up at any intensity was not significantly different from that without specific warm-up. The intracellular pH during the criterion exercise after specific warm-up at 30% or 40% MVC was significantly higher than that without specific warm-up. CONCLUSIONS: These results indicate that mild warm-up exercise could inhibit the development of intracellular acidosis during subsequent intense exercise. PMID- 11034433 TI - Physical, hematological, and exercise response of collegiate female equestrian athletes. AB - BACKGROUND: Physiological testing is extensively used to assess current physical status, target strength/deficiencies, and determine predisposition to injury in athletes. No studies exist regarding these issues on equestrian athletes. The purpose of this study was to quantify the physical, hematological, and exercise response of female equestrian athletes in order to provided greater insight into the health fitness of this unique competitor, and to compare results to other better studied sport athletes. METHODS: Following written informed consent, physiological assessments were performed on 24 collegiate female equestrian athletes (23.6+/-1.8 yrs; ht = 161.8+/-5.0 cm; wt = 64.9+/-9.3 kg) to quantify aerobic power (VO2max, VEmax, Timemax), anaerobic power (peak power, total work output, fatigue index), body composition (%BF, LBM), muscular strength (curl-ups, reverse sit-ups, pushups, handgrip strength), blood chemistries, and coronary risk profile. RESULTS: Data indicated that mean (+/-SD) LBM (49.0+/-4.5 kg) and BMI (24.8+/-1.7 wt/ht2) fell within reported athletic norms for females. Percent body fat (24.5+/-6.0%), however, was above average. Mean VO2max (33.9+/-4.5 ml/kg/min), treadmill time (10:06+/-:36 min:sec), and VEmax (90.3+/-16.0 l/min) were lower than predicted values. Observed peak power (5.3+/-0.9 w/kg), total work output (315.9+/-48.1 j/kg), fatigue index (66.9+/-7.9%), and handgrip strength (27.8+/-6.6 kg) were also lower than established norms for young females. Mean curl-up, reverse sit-up, and pushup responses (56.5+/-15.8, 36.6+/ 13.2, 32.1+/-10.6, reps/min, respectively) were considered average to above average for this age group. Mean resting blood chemistry values were within normal ranges. When analyzing the blood pressure, body composition, and serum lipid data, results indicate an average to low risk for coronary heart disease in this population. Wide variation in lipid profiles in this group as compared to athletes in other sports is a concern. CONCLUSIONS: Although working with equine poses a constant danger, the lack of adequate physical conditioning of the equestrian may be a contributing factor in the growing number of injuries. When compared to female athletes in other sports, exercise performance was found to be lower. Equestrian athletes need to supplement conventional sport activity with traditional aerobic and anaerobic training regimens. PMID- 11034434 TI - Changes in cortisol and testosterone levels and T/C ratio during an endurance competition and recovery. AB - BACKGROUND: To describe the evolution of cortisol and testosterone levels and testosterone/cortisol (T/C) ratio in long-distance runners during a relay competition and during the three days following the competition. METHODS: Two teams of four relayers (one male, one female) took part into this six-hour relay race. Hormonal control during the race was made possible thanks to saliva sampling during rest periods at each relay. The runners were subelite runners, (VO2max = 67.0 ml x min(-1) x kg(-1) in males and 56.8 ml x min(-1) x kg(-1) in females). RESULTS: During the race, cortisol levels reached approximately 1.5 fold basal levels. These levels remained high till late evening, (higher than morning values, when normal resting levels are 4 to 6-folds lower). Surprisingly, wakening levels during the following days were lower than resting levels. Testosterone did not vary in females; then, male values only are reported. During the race they decreased gradually and remained low till night. During the following three days, testosterone levels were higher than resting day levels. The T/C ratio amplifies these variations: low during the race till retiring, (currently associated with a catabolic tendency) and reversely high during the following three days (associated with a high anabolic tendency). CONCLUSIONS: As expected, a catabolic tendency occurs during a long distance run (increase in cortisol level followed by a drop in testosterone level). More surprising is the high anabolic tendency noted during the recovery period (low cortisol and high testosterone levels). PMID- 11034435 TI - Cardiorespiratory and metabolic responses during straight and bent knee cycling. AB - BACKGROUND: This study examined the influence of knee angle on the cardiorespiratory system loading during submaximal and maximal stationary cycle ergometry. METHODS: Experimental design and participants: eighteen untrained women (age: 21+/-1.88 years, weight: 57+/-5.75 kg, height: 165+/-5.03 cm, values are mean+/-SD) volunteered as subjects and underwent two-cycle ergometer incremental (Jaeger ER900) tests: 1) straight knee (180 degrees), 2) bent knee (140 degrees). MEASURES: oxygen uptake (VO2), ventilation (VE) and respiratory exchange ratio (RER) were measured continuously during each test using an open circuit spirometry and blood lactate concentration was determined by means of an enzymatic method. RESULTS: Comparing cycling with "straight knee" to cycling with "bent knee" at 50 W, heart rate (HR), V(E) and VO2 were significantly higher (10.6%, 12.5%, 17.8%). At 100 W, blood lactate was significantly lower (10.8%) while VO2 and RER was higher (5.5%, 7.1%). During maximal exercise, the total exercise time was significantly longer (11.2%) and VE, VO2 and HR were significantly higher during cycling with "straight knee" compared to cycling with "bent knee". No significant difference in peak lactate was evident between the two sitting positions. CONCLUSIONS: The results of this study indicate that cycling with bent knee requires lower oxygen uptake while pedaling with straight knee is the only way to reach VO2max during cycle testing, since the cardiorespiratory system is fully taxed. PMID- 11034436 TI - Echocardiographic findings in professional league soccer players. Effect of the position of the players on the echocardiographic parameters. AB - BACKGROUND: The purpose of the study was to compare the echocardiographic parameters of soccer players to healthy controls and to assess the effect of the position of the soccer players on the echocardiographic findings. METHODS: M Mode 2D and Doppler echocardiographic examination were carried out on 83 professional league soccer players and 52 healthy controls. RESULTS: Soccer players had increased interventricular septum (1.14+/-0.13 cm vs 0.99+/-0.17 cm, p<0.001) left ventricular posterior wall (1.08+/-0.16 cm vs 0.91+/-0.13 cm, p<0.001) thickness, increased left ventricular diastolic diameter (5.24+/-0.40 cm vs 4.88+/-0.43 cm, p<0.001), volume (133+/-23 ml vs 113+/-22 ml, p<0.001) and increased left ventricular mass index (142+/-28 g/m2 vs 103+/-23 g/m2, p<0.001) compared to controls. Soccer players had greater mitral E wave to A wave ratio (2.08+/-0.53 vs 1.65+/-0.43, p<0.001) compared to controls. Soccer players were subgrouped according to their position in the play as goal keepers, defensive, midfielder, and offensive players. Comparison among defensive, midfield and offensive players revealed subtle differences between defensive and midfield players. Right ventricular dimensions were higher in goal keepers compared to midfielders due to the greater body surface area of the goal keepers (2.8+/-0.4 cm vs 2.4+/-0.5 cm, p<0.05). Left ventricle end diastolic dimension were similar between groups but when these dimensions were corrected for the greater height of the defensive players the difference between midfields and defensive players became significant (3.05+/-0.18 vs 2.89+/-0.22, p=0.05). On Doppler flow parameters the ratio of peak E wave to A wave velocity (2.27+/-0.55 vs 1.84+/ 0.36, p<0.05) was increased in midfield players compared to defensive players. CONCLUSIONS: It was concluded that soccer players had greater left ventricular wall thickness, volume and mass compared to controls. But the effect of the position of the players on the measured cardiac dimensions were minimal. These subtle findings were explained by the leveling effect of the playing system and training, and lack of physical training during the previous month. PMID- 11034437 TI - Comparison of exertional indices following moderate training in collegiate athletes. AB - BACKGROUND: To better understand the inflammatory response to two divergent training modalities: running and weight training. METHODS: EXPERIMENTAL DESIGN: prospective, repeat measures, observational study of subjects over the course of 8-10 weeks of training. Data analysis was consonant with this design and included a repeat measures ANOVA with multiple posthoc comparisons. The level of significance was set at 0.05. SETTING: collegiate athletic training facilities. PATIENTS: twenty-two subjects comprised the study group. These included both male and female collegiate athletes, ages 17-22 years of age with a mean height of 178.6 cm, a mean weight of 77.5 kg and a mean lean body mass of 67.2 kg. Exclusion criteria consisted of hemophilia or chronic conditions requiring anti inflammatory medication. INTERVENTIONS: training for runners (n=11) consisted of multiple 5-km runs throughout the course of a 60-75 minute workout. For a comparable time period, weight trainers participated in multiple joint, total body strengthening workouts. MEASURES: four exertional indices were examined. These included serum levels of creatine kinase, lactate, Interleukin-6, and C reactive protein. Assessments of these markers were made immediately before and after each training session. Three sessions were monitored over an 8-10 week period. RESULTS: The data show that moderate exertion is accompanied by modest increases in these markers. Mean serum values did not exceed 125 U/L for serum creatine kinase, 5 mM/I for lactates, and 2.6 pg/ml for IL-6. C-reactive protein was sporadically detectable in weight lifters only. These increases were detected immediately postexercise. Each training modality produced a unique profile for creatine kinase and IL-6, suggestive of a training effect. CONCLUSIONS: Overall, the levels for all exertional markers reported herein are well below those reported for exhaustive exercise. As consistent exhaustive efforts can catalyze overtraining syndromes, periodic assessments of these exertional indices could help to document the appropriateness of the training stimulus. PMID- 11034438 TI - Investigation of anthropometric and work-rate profiles of elite South American international soccer players. AB - BACKGROUND: The aim of the current investigation was to determine the movement profiles of elite South American soccer players during international competition and examine the relationship between anthropometric profile and work-rate variables. METHODS: Seventeen full-time professional soccer players were filmed while competing for their countries. Anthropometric profiles were obtained for eleven of these players. Six full-time professional players from the English Premier League were also filmed for comparative purposes. RESULTS: The South American international players covered significantly less (p<0.05) total distance during match-play than English Premier League players (International, 8638+/-1158 m; English Premier League, 10104+/-703 m). The total distance covered during the second half of the game was significantly reduced (p<0.05) compared to the first half distance for both groups of players (mean+/-SD first half 4605+/-625 m; mean+/-SD second half 4415+/-634 m). The data for both groups of players were combined to evaluate positional differences in the workrate profile. Midfield players covered a significantly greater (p<0.05) distance than forward players (midfield, 9826+/-1031 m; forwards, 7736+/-929 m) and defenders covered a greater (p<0.05) distance jogging backwards than forward players (defenders, 276+/-155 m; forwards, 68+/-25 m). Forwards sprinted a greater distance (p<0.05) than defensive players (defenders, 231+/-142 m; forwards, 557+/-142 m). Mean somatotype was a balanced mesomorph (2-5 1/2-2). Body mass and muscle mass was related to the total distance covered (r=0.43, r=0.53, p<0.05). CONCLUSIONS: Based on these data, it seems that an individual's work-rate profile is dependent upon the type of competition and the playing position. Relationships between anthropometric profile and work-rate are complex due to the interaction between the variables that determine work-rate. PMID- 11034439 TI - Adiposity does not hinder the fitness response to exercise training in obese women. AB - BACKGROUND: This study examined how sedentary obese (OB) and normal weight (NW) women respond to exercise training; and if fitness levels of the OB and NW are comparable, in spite of differences in adiposity. METHODS: Sedentary OB (n=46, 48.5+/-1.5 yrs, BMI=35.9+/-0.8; mean+/-SEM) and NW (n=43, 48.3+/-1.7 yrs, BMI=21.7+/-0.2) women participated in a 14-week fitness program. RESULTS: There were no group differences in exercise adherence. No changes for either group were found for body weight or composition. Both groups improved similarly in aerobic fitness (VO2max), muscular strength (grip strength), muscular endurance (modified push-up), and flexibility (sit and reach). However, since norms for fitness are generally expressed relative to body weight (e.g. VO2max, ml x kg(-1) x min(-1); grip strength ratios), the OB women continued to be classified as unfit after exercise training, and categorized below the NW women in spite of having absolute scores for VO2max (OB=2.1+/-0.1, NW=1.8+/-0.1 l x min(-1)) and grip strength (OB=65.1+/-1.5, NW= 58.6+/-1.5 kg) that were higher than the NW women. CONCLUSIONS: The data suggest that OB women respond in a similar manner as NW women to exercise; that weight loss is not necessary for improved fitness; and that the OB are not less fit than the NW, but that low fitness scores for the OB are simply a reflection of the absolute scores being deflated because they are proportioned to body weight. PMID- 11034440 TI - Body composition measurement in highly trained male athletes. A comparison of three methods. AB - BACKGROUND: The purpose of the present study was to evaluate the differences in the body composition estimate of highly trained male athletes by three different methods: dual-energy X-ray absorptiometry (DXA), bioelectrical impedance analysis (BIA) and skinfold thickness measurement (SFT). METHODS: The study occurred during a non-intensive training period. The measurements were performed at the Human Physiology laboratory. PARTICIPANTS: forty-three male athletes (19 waterpolo, 9 judo, 15 karate) who exercised regularly at least three hours per day, six days per week. MEASUREMENTS: DXA, BIA and SFT measurements of percentage fat mass (%FM) and the amount of fat-free mass (FFM) were performed. STATISTICAL ANALYSES: the different %FM estimates by the three methods were compared using a one-way ANOVA, with posthoc Bonferroni test when values were significantly different. The same test was used to compare FFM values. The coefficient of variation (CV%=100xSD/mean) was calculated for each methods. Methodological differences among the various methods were analysed with the method of Bland Altman. RESULTS: ANOVA test showed significant differences among the three methods. In particular, DXA significantly (p<0.001) overestimated %FM (12.4+/ 4.1%) and underestimate FFM (67.0+/-6.9 kg) respect to SFT (7.8+/-0.9 % and 71.0+/-8.2 kg) and BIA (9.9+/-1.4% and 69.4+/-7.9 kg). Also Bland-Altman comparison among the estimates indicated wide differences between methods. CONCLUSIONS: DXA provides different body composition estimates than those derived from skinfold thickness measurement and BIA, so that the methods should not be used interchangeably. PMID- 11034441 TI - Detection of HCV and GBV-CHGV RNA in peripheral blood mononuclear cells of patients with chronic type C hepatitis. AB - Hepatitis C virus (HCV) and hepatitis G virus (HGV) viraemia were investigated by RT-PCR protocols in peripheral blood mononuclear cells (PBMC) of 22 patients with chronic type C hepatitis. Samplings were at basal and 4-8 months after a 12 month period of treatment with interferon-alpha. A plus strand of HCV in PBMC was detected in 8 of 21 patients (38%) (p <0.05; chi2 test) with a lack of response to therapy; a minus strand was detected in 10% of chronic type C hepatitis and 25% of the patients harboured HCV RNA in PBMC. The association with a response was nearly significant (p <0.1; chi2 test). GBV-C/HGV RNA was detected in the serum of 9 of 21 (43%) patients and in PBMC of 20% of the patients viraemic. Genomic sequences of GBVC/HGV in PBMC were found, but further investigation is needed to assess the findings reported for HCV. PMID- 11034442 TI - Endotoxins stimulate generation of superoxide radicals and lipid peroxidation in blood platelets. AB - Lipopolysaccharide (LPS, endotoxin) is an important structural constituent of the membrane of gram-negative bacteria with a wide range of biological effects. It can activate blood platelets. The purpose of present study was to determine the direct effect of endotoxins from Proteus mirabilis, differing significantly in their composition, on the generation of superoxide radicals and thiobarbituric acid reactive substances (TBARS) in blood platelets. Superoxide radicals were measured by means of superoxide dismutase-inhibitable reduction of cytochrome C. The TBARS determination (malonyldialdehyde) was used as a marker of endogenous arachidonate metabolism and thromboxane A2 synthesis. Results demonstrate that three endotoxins (LPS S1959, LPS R110, LPS R45) after 2 min of action, even at the lowest concentration (0.03 microg/10(8) platelets) stimulated the generation of TBARS and release of superoxide radicals. All LPS contain lipid A as a component but differ in their chemical composition in the polysaccharide part. It is suggested that the observed effects of LPS on blood platelets are attributable to their lipid A portion. PMID- 11034443 TI - Homogeneity of Trypanosoma evansi isolates from domestic and sylvatic mammals from the Pantanal of Mato Grosso. AB - 'Mal de Cadeiras' is a disease which causes great mortality in horses in the Pantanal Matogrossense region, Brazil. The agent of this disease is Trypanosoma evansi, a kinetoplastid flagellate which belongs to the Trypanosomatidae family, classified into the Salivarian section. Transmission occurs mechanically by haematophagous Diptera, mainly by Stomoxys sp. and Tabanus sp. and vampire bats. Outbreaks of Mal de Cadeiras in horses result in economic losses, thus limiting their use in cattle raising. Ten isolates of T. evansi recently derived from coati (Nasua nasua, Carnivora, Procyonidae), horses and dogs were compared, using schizodeme analyses from DNA digested by the restriction enzyme Hin fl. The results showed similar electrophoretic profiles for all isolates from wherever the host came. Homogeneity of isolates from domestic and sylvatic animals suggested two hypotheses: (1) the parasites circulated in only one transmission cycle;, and (2) independent cycles were not established in sufficient time to modify the molecular profiles of the isolates. PMID- 11034444 TI - The minimum inhibitory concentration of oral antibacterial agents against cariogenic organisms. AB - The minimum inhibitory concentrations (MIC) of eight common dental antibacterial agents against three genera of bacteria which have been implicated in dentine caries, namely streptococci, lactobacilli and actinomycetes were investigated. The ultimate aim was to determine the most appropriate antibacterial agent which could be added to dental restorative materials for filling cavities where there was residual dentine caries. The antibacterial agents tested were chlorhexidine diacetate, chlorhexidine dihydrochloride, chlorhexidine gluconate, benzalkonium chloride, cetrimide, cetylpyridinium chloride, thymol and sodium hypochlorite. Thymol and sodium hypochlorite did not inhibit microbial growth at any of the concentrations tested. For the active antibacterial agents tested the MIC values against lactobacilli and streptococci were 0.25 microg/ml to 8.0 microg/ml and for actinomycetes 0.125 to 8.0 microg/ml. These results illustrate the wide spectrum of sensitivity of caries associated bacteria against dental antibacterial agents. From the MIC values alone, it is difficult to recommend which of the active antibacterial agents would be most effective in eliminating cariogenic organisms. PMID- 11034445 TI - Distribution of diaminopropane, putrescine and cadaverine in Haemophilus and Actinobacillus. AB - Cellular levels of diaminopropane, putrescine and cadaverine, and decarboxylase activities to produce these diamines in six species (16 strains) of Haemophilus and four species (5 strains) of Actinobacillus belonging to the family Pasteurellaceae of the gamma subclass of the class Proteobacteria, were determined by high performance liquid chromatography (HPLC). Diaminopropane was ubiquitously distributed within all Haemophilus and Actinobacillus species, and L 2,4-diaminobutyric acid decarboxylase activity was detected in them. Putrescine and ornithine decarboxylase activity were found in H. aphrophilus, H. parainfluenzae and H. influenzae (type a, b, d, e and f except for type c) but not detected in H. aegyptius, H. parahaemolyticus, H. ducreyi and Actinobacillus species. Cadaverine occurred in H. aphrophilus, H. aegyptius, H. influenzae, H. parainfluenzae, A. actinomycetemcomitans, A. equuli and A. lignieresii, whereas their lysine decarboxylase activity was scarcely detected. Cadaverine was not found in H. parahaemolyticus, H. ducreyi and A. suis. The diamine profile serves as a phenotypic marker for the chemotaxonomic classification of the family Pasteurellaceae. PMID- 11034446 TI - The effect of azidothymidine on germ tube formation in Candida albicans. AB - Candida albicans have a marked propensity to cause infections in AIDS patients. A virulent trait of C. albicans is the yeast-hypha transition (Y-->H) which is influenced, in vitro and in vivo, by several factors. Since azidothymidine (AZT) is used in HIV-positive patients, the effect, in vitro, of different concentrations of AZT on C. albicans Y-->H transition was evaluated. C. albicans isolated from HIV-negative and HIV-positive patients were used and strains of C. tropicalis isolated from HIV-positive patients were also tested. AZT concentrations from 0.01 microg/ml to 10 microg/ml did not have any influence on the Y-->H transition, whereas 100 microg/ml AZT significantly inhibited the germ tube formation. AZT did not influence the formation of pseudohyphae in C. tropicalis. It is suggested that C. albicans infection observed in HIV-positive patients was not influenced by AZT therapy, because at currently used dosages, the Y-->H transition was not expected to increase. PMID- 11034447 TI - Anti-hepatitic C virus antibodies hidden in circulating antibody/antigen aggregates in HCV-RNA positive patients. AB - The aim of this study was to determine whether antibodies to HCV can be hidden in immunocomplex aggregates in anti-hepatitis C virus (HCV) negative, HCV-RNA positive patients and whether their presence could be related to HCV viral load or HCV genotype. Sera (23 in toto) from patients with elevated alanine aminotransferase (ALT) levels and negative for anti-HCV but positive for HCV-RNA and the immunocomplex aggregates (precipitate with PEG 6000 and glycine 1 M) were studied. The sera were treated using a rapid, simple new ELISA which disrupted the immunocomplex aggregates. Sera from ten patients were tested anti-HCV positive after immunocomplex disruption. No correlation with age, sex, ALT level, viral load or HCV genotype was observed. In some patients anti-HCV antibodies were hidden in circulating antibody/antigen complexes which could be dissociated with a simple, inexpensive and rapid protocol; therefore it can provide a valuable addition to the diagnosis of HCV infection and it may prevent some cases of post-transfusion hepatitis. PMID- 11034448 TI - Hepatitis C virus and Epstein-Barr virus: dual infection or atypical antibody response. AB - Positive serological reactions against hepatitis C virus (HCV) appeared in the course of Epstein-Barr virus (EBV) infectious mononucleosis. In 429 consecutive patients with high levels of transaminases, 28 patients with EBV primary infection were found. The presence of anti-HCV antibodies and HCV RNA was studied in these subjects. In seven patients anti-HCV antibodies (C33 and C22c RIBA bands) were detected, but all were polymerase chain reaction (PCR) negative. These results may have been due solely to a HCV infection or were an atypical response to HCV in the course of infectious mononucleosis. PMID- 11034449 TI - 'Treat the patient, not the EEG'? PMID- 11034450 TI - Acquired aphasia in children after surgical resection of left-thalamic tumors. AB - Five children (three males, two females; four right-, one left-handed; age range 6 to 14 years) who developed aphasia after gross-total excision of left predominantly thalamic tumors are reported. Three patients had Broca aphasia, one had mixed transcortical aphasia, and one patient had conduction aphasia. In the months after surgery, three children improved while receiving radiation and/or chemotherapy, although none recovered completely. Two patients with malignant tumors developed worsening aphasia when the tumor recurred, and later died. Two of three patients tested had visuospatial difficulties in addition to language deficits. Attention and executive functioning were affected in three of three patients tested. Memory, verbal and/or visual functioning, were affected in four of four patients tested. Both patients who were tested showed transient right hemineglect. Two of two patients tested were probably apraxic. The wide range of deficits in these children highlights the importance of the thalamus and other subcortical structures in developing cognition. PMID- 11034451 TI - Early posttraumatic seizures in non-accidental head injury: relation to outcome. AB - To document the characteristics of early posttraumatic seizures (EPTS) in non accidental head injury (NAHI), and examine their relation with outcome, a retrospective study was carried out. All children with NAHI admitted to the Royal Hospital for Sick Children, Edinburgh, since 1981 were identified. The characteristics of EPTS, EEG, and outcome were noted. Forty-four cases were identified. The average age of children at presentation was 5.9 months. Thirty two of these children had EPTS. The median length of follow-up was 3 years. The mortality rate was six in 44 (14%). The neurodevelopmental outcome correlated significantly with the presence and severity of EPTS (Tau=0.317,p=0.017). Of survivors, 22% developed late posttraumatic epilepsy; the outcome in those with epilepsy was significantly worse than those without (p<0.0001). It was concluded that the severity of the primary brain injury dictates the severity of the EPTS and neurodevelopmental status at follow-up. PMID- 11034452 TI - Benign partial epilepsy of childhood: a longitudinal neuropsychological and EEG study of cognitive function. AB - The study combined prospective neuropsychological and EEG results of 22 children presenting with typical benign partial epilepsy with rolandic spikes (n=19) and occipital spikes (n=3). The aims were to assess the types of cognitive problems which may be encountered in this population, to evaluate the course of cognitive and learning capacities during the active phase of epilepsy, and to see if there was a correlation with paroxysmal activity on the EEG. Average age at entry in the study was 8.4 years and each child was seen two to four times over a period of 1 to 3 years. EEGs showed persistent spike foci in most cases that worsened in three cases, but there were no continuous spike-waves during sleep. No child had persistent stagnation, marked fluctuations, or a regression in cognitive abilities. Of 22 children, 21 had average IQ (>80). Eight children had school difficulties requiring special adjustment. No single cognitive profile was identified. Four children had delayed language development and eight children had transient weak scores in one isolated domain (verbal, visuospatial, memory) which improved or normalized during the course of the study with concomitant EEG improvement or normalization. In two of the three children with aggravation of the paroxysmal EEG activity, clinical changes were documented. A proportion of children with typical benign partial epilepsy with rolandic spikes showed mild, varied, and transient cognitive difficulties during the course of their epilepsy, and in most cases this probably had a direct relation with the paroxysmal EEG activity. PMID- 11034453 TI - Maternal expressed emotion and treatment compliance of children with epilepsy. AB - This study investigated the association between family relationships and compliance in the treatment of childhood epilepsy. It was a prospective study of 21 families with a child who had epilepsy attending Central Middlesex Hospital, London. There were 13 boys and eight girls, with a mean age of 12.0 years (SD 2.9) at initial assessment. Mothers were interviewed for assessment of expressed emotion as a measure of parent-child relationships. Assessment of the mothers' adjustment using the General Health Questionnaire (Goldberg 1978), and psychological adjustment of the children using the Rutter Scales (Rutter et al. 1970a), were completed by mothers and teachers. Reassessment was 3 to 4 years after initial contact, including a paediatric case-note review to assess clinic attendance and overall treatment compliance. Significantly more of the group who had a good level of compliance had recovered from epilepsy at follow-up. Good treatment compliance was found to be associated with less maternal hostility and criticism. Children and mothers in the good compliance group had fewer psychiatric symptoms. Poor treatment compliance and the associated psychological disturbances suggest that assertive paediatric and psychosocial intervention may be needed for some children with epilepsy. PMID- 11034454 TI - Preliminary validation of an observational pain checklist for persons with cognitive impairments and inability to communicate verbally. AB - To obtain a preliminary validation of the Non-Communicating Children's Pain Checklist for individuals with an inability to communicate verbally, 32 caregivers of individuals with cognitive impairments aged 3 to 44 years retrospectively completed the Non-Communicating Children's Pain Checklist and rated item usefulness (0 to 10). In the second part of the study 33 caregivers completed the Checklist after two painful (e.g. burns, falls, surgery), one distressful (e.g. unwanted grooming, feared animal/noise), and one calm event (e.g. watching television). Checklist scores did not correlate with the age, sex, or physical limitations of individuals with an inability to communicate verbally. In the first part of the study Cronbach's alpha was 0.66; all mean usefulness ratings exceeded 5 out of 10. In the second part of the study after four items were removed, Cronbach's alpha was 0.79. Checklist scores during pain correlated with numerical ratings of pain intensity did not differ between the two pain events, and differed significantly from calm scores. Differences in Checklist scores during pain and distress were found for two subscales. The Checklist exhibits internal consistency, and preliminary evidence suggests it can detect pain and is reliable over time. PMID- 11034455 TI - Non-invasive technique for assessment and management planning of oral-pharyngeal dysphagia in children with cerebral palsy. AB - The non-invasive Exeter Dysphagia Assessment Technique (EDAT) was evaluated as a method of assessing the aetiology of dysphagia in children with cerebral palsy (CP). Data were collected from a group of 20 typically developing children (nine girls, 11 boys; age range 7 to 14 years) for comparison with 125 dysphagic children with CP (81 boys, 44 girls; age range 1 to 18 years). The swallowing mechanism has been separated into physiological phases: anticipatory, delivery, oral transit, and oral-pharyngeal. Normal or abnormal function in each phase was recorded and the common causes of any impaired phase were considered, starting with generalized possibilities before focusing on specific parts of swallowing physiology. Data from 125 dysphagic children with CP show marked differences from the data for the typically developing children. Interpreting individual results was valuable in assisting the assessment team to formulate management strategies; two examples are presented. The technique appears to provide a cost-effective, non-invasive, and valuable clinical tool. PMID- 11034456 TI - Changes in the incidence of childhood autism and other autistic spectrum disorders in preschool children from two areas of the West Midlands, UK. AB - The incidence of childhood autism and other autistic spectrum disorders (ASDs) in preschool children was determined for two areas of the West Midlands between 1991 and 1996. Children diagnosed before the age of 5 years and residing within the study areas at diagnosis were detected from the records of four child development centres. The incidence rate per 10,000 children per year for the combined areas was 8.3 for all children with ASDs, 3.5 for classical childhood autism (CA), and 4.8 for other ASDs. Rates were similar in both areas, despite differences in social deprivation and proportions of ethnic minorities. While rates for classical CA increased by 18% per year, a much larger increase (55% per year) was seen for 'other ASDs', suggesting that clinicians are becoming increasingly able and/or willing to diagnose ASDs in preschool children. PMID- 11034457 TI - 3-phosphoglycerate dehydrogenase deficiency in a patient with West syndrome. AB - 3-phosphoglycerate dehydrogenase deficiency is a severe but treatable disorder of serine synthesis, first described in 1996 (Jaeken et al. 1996a). The patient presented with West syndrome, severe psychomotor delay, failure to thrive, microcephaly, atypical ocular movements, and pyramidal signs. Treatment with oral L-serine abolished seizures and improved psychomotor development, hyperexcitability, head growth, cortical and subcortical hypotrophy, and hypomyelination of the brain on MRI scans. 3-phosphoglycerate dehydrogenase deficiency is a treatable congenital error that probably leads to West syndrome. PMID- 11034458 TI - Evidence of the effects of intrathecal baclofen for spastic and dystonic cerebral palsy. AACPDM Treatment Outcomes Committee Review Panel. PMID- 11034459 TI - Input and outcome, numerators and denominators. PMID- 11034460 TI - Melatonin as a sedation substitute for diagnostic procedures. PMID- 11034461 TI - Presentation and outcome of sacral agenesis: 20 years' outcome. PMID- 11034462 TI - Surgical management of liver injuries in adults--current indications and pitfalls of operative and non-operative policies: a review. PMID- 11034463 TI - Primary tuberculosis of the breast in Qatar: ten year experience and review of the literature. AB - OBJECTIVE: To present our experience of isolated tuberculosis of the breast, 1988 98. DESIGN: Retrospective study of case notes and radiographs. SETTING: Hamad General Hospital, Qatar. SUBJECTS: 13 multiparous women with tuberculosis of the breast. MAIN OUTCOME MEASURES: Objective confirmation of diagnosis and adequacy of treatment. RESULTS: We found an overall incidence of histologically confirmed tuberculosis to be 0.4%/year. All patients presented with a lump, 2 had nipple discharge and one had a palpable axillary node on the same side. 7 were treated by excision biopsy, 3 by incision and drainage, and 3 had fine needle aspiration (FNA) as their only procedure. All diagnoses were confirmed histologically. 2 patients developed recurrences during treatment but these were successfully treated. CONCLUSION: Although the incidence of the disease is low, the diagnosis should be suspected in young multiparous women with a breast lump in whom malignancy has been excluded. The minimum of surgical intervention (incision or excision biopsy) together with antituberculous drugs seems to be the most successful treatment. PMID- 11034464 TI - Role of intraoperative ultrasound of the liver with improved preoperative hepatic imaging. AB - OBJECTIVE: To find out whether intraoperative ultrasound (IOUS) of the liver should still be considered a routine procedure for patients having hepatic resections as their preoperative investigation has become so much better with the standard use of triphasic spiral computed tomograms (CT). DESIGN: Retrospective study. SETTING: University medical centre, Utrecht, The Netherlands. PATIENTS: 26 consecutive patients undergoing liver resection. MAIN OUTCOME MEASURES: How much additional information was given by IOUS, and its influence on operative strategy; how IOUS facilitated the hepatic resection. RESULTS: IOUS revealed additional information over triphasic CT in 13/26 patients, which led to a change in the surgical procedure in 4 (15%). For all 4 patients the change in surgical approach proved to be correct as judged by histology and clinical outcome. In 12 patients IOUS facilitated the resection by providing helpful three-dimensional anatomical insight. CONCLUSION: Despite recent improvements in preoperative investigation, IOUS should still be recommended as a routine procedure in patients having hepatic resections. In addition to aiding the ultimate decision about which part to resect, it also gives insight into the hepatic anatomy. PMID- 11034465 TI - Management of residual cavity after partial cystectomy for hepatic hydatidosis: comparison of omentoplasty with external drainage. AB - OBJECTIVE: To investigate the role of two approaches to the operative treatment of hepatic hydatid cysts. DESIGN: Prospective study. SETTING: University and teaching hospital, Turkey. SUBJECTS: 108 patients with single uncomplicated hydatid cysts who were operated on in two clinics between 1990 and 1995. INTERVENTION: Introflexion and omentoplasty or external drainage after partial cystectomy in single uncomplicated hydatid cysts more than 5 cm in size. MAIN OUTCOME MEASURES: Morbidity, mortality, and hospital stay. RESULTS: The median hospital stay after introflexion and omentoplasty was 8 days (range 3-15), which was significantly shorter than that after external drainage (12 days, range 7 20). There were 2/35 postoperative complications in the former group compared with 17/73 in the drainage group (p = 0.03). There was one death after introflexion and omentoplasty. CONCLUSION: Introflexion and omentoplasty after partial cystectomy for a single uncomplicated hydatid cyst caused significantly fewer complications than external drainage, and patients left hospital sooner. PMID- 11034466 TI - Systemic inflammatory response in acute cholangitis and after subsequent treatment. AB - OBJECTIVE: To measure the concentrations of endotoxin and inflammatory mediators during an attack of acute cholangitis and see what effect endoscopic treatment had on these mediators. DESIGN: Prospective study. SETTING: University teaching hospital The Netherlands. SUBJECTS: Ten patients with acute cholangitis. INTERVENTIONS: Measurements were made during the attack and 1 week after endoscopic treatment. MAIN OUTCOME MEASURE: Changes in clinical variables, and severity of biliary obstruction. Concentrations of endotoxin, cytokines, and endotoxin binding proteins, in plasma. RESULTS: The causes of cholangitis were obstructed endoprosthesis (n = 4) and stones (n = 6). The median bilirubin concentration during the attack was 70.0 micromol/L (range 14-156) and 14.5 micromol/L (range 9-80) after treatment (p < 0.05). Median (range) plasma endotoxin concentrations were 3.6 pg/ml (3.2-107) and 3.6 (2.4-5), respectively. Concentrations of cytokines were high during the acute attack and significantly lower after treatment: median tumour necrosis factor (TNF) fell from 44.6 pg/ml (range 1.2-403) to 7.3 (0-53); soluble TNF receptor p55 from 4.9 ng/ml (2.7-13.8) to 3.6 (1.4-8.2) and TNF receptor p75 from 11.6 ng/ml (7.1-40.6) to 8.1 (2.9 31.3); interleukin 6 (IL-6) fell from 690 pg/ml (34.1-4594) to 8.2 (0-39.3), IL-8 from 226.2 pg/ml (31.6-712.7) to 21.4 (4.2-63.5) and IL-10 from 33.4 pg/ml (2.7 5605) to 4.7 (0-16.7) (p < 0.03). Values for lipopolysaccharide binding protein and soluble CD14 also fell significantly (p < 0.01) from 86.5 (43.4-200) to 21.5 (11.3-37.5) and from 200 (59-200) to 47.8 (0.47-200), respectively. The concentration of bactericidal permeability increasing protein did not change significantly, being 7.1 (2-18.9) during the acute attack and 4.6 (0.8-17.7) a week later. CONCLUSION: There is a considerable systemic inflammatory response during cholangitis, which is dramatically reduced one week after endoscopic treatment. PMID- 11034467 TI - Pooling of prognostic studies in cancer of the pancreatic head and periampullary region: the Triple-P study. Triple-P study group. AB - OBJECTIVE: Development of a prognostic tool for patients with unresectable pancreatic cancer to distinguish between with low or high probabilities of survival 3 to 9 months after diagnosis. DESIGN: Data about individual patients from five studies were pooled. A multivariate proportional hazards model with time-dependent covariates was developed, including age, sex, and metastases. An extended model was developed on a subset of patients, including weight loss, pain, and jaundice at diagnosis. SETTING: Multicentre study, The Netherlands, Norway, USA, UK, and Canada. SUBJECTS: 1020 patients with unresectable pancreatic cancer. MAIN OUTCOME MEASURES: Prediction of prognosis. RESULTS: Patients with metastases, pain, or weight loss at diagnosis had a significantly poorer prognosis than the others. Older men had a worse prognosis than younger men, while older women had a better prognosis than younger ones. Patients with jaundice had a relatively good prognosis. Differences in survival among the studies were incorporated in a prognostic score chart. CONCLUSION: The prognostic score chart can be used to select patients with relatively low expectation of survival for endoscopic palliation, and patients with relatively high expectation for surgical palliation. PMID- 11034468 TI - Medical or surgical treatment for chronic gastrooesophageal reflux? A systematic review of published evidence of effectiveness. AB - OBJECTIVE: To compare the effectiveness of medical (antacids, histamine antagonists and proton pump inhibitors) and surgical (fundoplication) treatment of chronic GORD. SUBJECTS: Patients with objective (endoscopic or pH) evidence of chronic reflux reported in 6 randomised trials and 3 cohort studies, 1966-1999. METHODS: Systematic review of comparative studies identified from electronic searches, citations, manual searches of journals, and correspondence with authors and experts. MAIN OUTCOME MEASURES: Improvements in prevalence or severity of symptoms, oesophagitis, pH reflux duration, lower oesophageal sphincter pressure, patients' satisfaction, and side-effects. RESULTS: Improved outcomes were more common after surgical than medical treatment with significant differences in objective outcomes in 5/6 randomised trials and in 2/3 cohort studies. Subjective outcomes (symptoms and patients' satisfaction) were also more common among surgical patients in all but one study that assessed them. Odds ratios for improvement with surgical rather than medical treatment ranged from 1.2 to 200, and numbers needed to treat ranged from 1.2 to 58, where these could be calculated. Studies were too heterogeneous for meta-analysis. CONCLUSIONS: In trials of chronic severe GORD, surgery is consistently more effective than medical treatment in relieving symptoms and objective oesophagitis, although omeprazole can give similar symptom relief with adjustment of the dose. PMID- 11034469 TI - Melatonin modulates mesenteric blood flow and TNFalpha concentrations after lipopolysaccharide challenge. AB - OBJECTIVE: To investigate the effect of various doses of melatonin on reduction in mesenteric blood flow (MBF) and increase in tumour necrosis factor alpha (TNFalpha) concentration caused by injection of lipopolysaccharide (LPS). DESIGN: University Hospital, Turkey. SETTING: Open experimental study. ANIMALS: 59 Swiss albino mice. INTERVENTIONS: Animals were injected with melatonin solvent or 1, 10, 100, or 500 mg/kg melatonin. Ten minutes later control animals were injected with saline, and the experimental group with LPS. MAIN OUTCOME MEASURES: Mesenteric blood flow and serum TNFalpha concentration. RESULTS: In control animals, 100 and 500 mg/kg melatonin reduced MBF. LPS reduced MBF in solvent, 1, and 10 mg/kg melatonin groups. The concentration of TNFalpha was considerably increased in the mice given LPS. Melatonin reduced this response significantly. CONCLUSION: In high doses melatonin directly reduces MBF. It has no protective effect on the LPS-induced decrease in MBF. In lower doses it blocks, but at higher doses reduces, LPS-induced TNFalpha production. PMID- 11034470 TI - Influence of glutathione metabolising enzymes in rats with gram-negative sepsis. AB - OBJECTIVE: To evaluate the role of glutathione metabolising enzymes in rats with Gram-negative sepsis. SETTING: University hospital, Sweden. ANIMALS: 61 male Sprague Dawley rats. INTERVENTIONS: Animals were divided into two groups, one of which was given tert-butyl-4-hydroxyanisole (BHA), a powerful inducer of glutathione metabolising enzymes. A gelatine capsule containing bacteria and adjuvant substances was placed in the abdomen. In one group it contained only adjuvant substances, and the controls underwent sham laparotomy. MAIN OUTCOME MEASURES: Activities of glutathione metabolising enzymes, and histological effect on pulmonary tissue. RESULTS: Activities of glutathione metabolising enzymes were reduced in lung tissue after the induction of sepsis. Pre-treatment with BHA increased enzyme activity and reduced the histological changes. CONCLUSION: Glutathione metabolising enzymes may have a role in sepsis, and pre-treatment with BHA seems to prevent histological changes in pulmonary tissue in septic rats. PMID- 11034472 TI - Coeliac disease diagnosed after pancreaticoduodenectomy. PMID- 11034471 TI - Single doses of FK506 and OKT3 reduce severity in early experimental acute pancreatitis. AB - OBJECTIVE: To find out if two immunomodulatory drugs used in organ transplantation (FK506 (tacrolimus) and OKT3 (Orthoclone) would reduce early inflammatory complications in experimental acute pancreatitis. DESIGN: Laboratory study. SETTING: University hospital, Germany. ANIMALS: 36 Balb/c mice. INTERVENTIONS: Pancreatitis induced by 7 intraperitoneal injections of cerulein 50 microg/kg at hourly intervals followed by FK506 0.32 mg/kg, OKT3 0.6 mg/kg, or 0.9% sodium chloride (controls) (n = 12 in each group). 12 hours after induction of pancreatitis the animals were killed. MAIN OUTCOME MEASURES: Serum amylase activity and interleukin-6 (IL-6) concentrations; histological damage to pancreas and lungs, apoptotic cells in pancreas; and myeloperoxidase activity in lungs. RESULTS: No animal died during the experiment. At 12h serum amylase activity and IL-6 concentrations were increased in all 3 groups, but highest in the OKT3 group. The pancreatic histological score, apoptosis, and inflammatory infiltration were lower in the two experimental groups than controls, but the degree of vacuolisation of acinar cells was similar. Packed cell volume was higher in the control than the experimental groups, and pulmonary damage and myeloperoxidase activity were less in the experimental groups than the controls. CONCLUSION: Single therapeutic doses of FK506 and OKT3 reduced the early severity of pancreatitis, pulmonary damage, and haemoconcentration in mice. Single doses of FK506 or OKT3 may therefore be effective in preventing the early complications of pancreatitis. PMID- 11034473 TI - Videolaparoscopic removal of a foreign body from the liver. PMID- 11034474 TI - The place of colostomy after late detection of a colonic injury and faecal peritonitis. PMID- 11034475 TI - Lateral internal sphincterotomy with haemorrhoidectomy for the treatment of prolapsed haemorrhoids. PMID- 11034476 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 11034477 TI - Notification that new names and new combinations have appeared in volume 50, part 4, of the IJSEM. PMID- 11034478 TI - Classification of aster yellows-group phytoplasmas based on combined analyses of rRNA and tuf gene sequences. AB - Seventy phytoplasma isolates, including 10 previously characterized reference strains, of the aster yellows group were examined by RFLP analysis of PCR amplified rDNA and RFLP and sequence analysis of the tuf gene. On the basis of rDNA restriction profiles, seven previously proposed 16S rDNA subgroups (16SrI-A, -B, -C, -D, -E, -F and -K) were recognized in the material examined. In addition, three new subgroups that differ in the RFLP profiles were identified and designated 16SrI-L, 16SrI-M and 16SrI-N. Of the two types of rDNA sequences used, an 1800 bp fragment comprising the entire 16S rRNA gene and the 16S-23S rDNA spacer region proved more suitable for AY-group phytoplasma differentiation than a 1240 bp fragment of the 16S rRNA gene. Many differences in the rDNA profiles between the subgroups could be explained by sequence heterogeneity of the two phytoplasmal rRNA operons. The subgroups delineated by RFLP analysis of a 940 bp tuf gene fragment are consistent with subgroups defined on the basis of rDNA sequences. However, subgroups 16SrI-D, -L and -M showed the same tuf gene restriction profiles as subgroup 16SrI-B. This result was confirmed by sequence analysis in which these subgroups differed slightly in their tuf gene sequence, when compared with members of subgroup 16SrI-B. On the basis of combined analyses of rDNA and tuf gene sequences and in view of pathological aspects, the taxonomic distinction of AY-subgroups 16SrI-A, -B, -C, -D, -E, -F, -K and -N appears to be substantial. PMID- 11034479 TI - Phylogeny of Bacillus sphaericus-like organisms. AB - The mesophilic round-spored bacteria embrace four species, namely Bacillus sphaericus, Bacillus fusiformis, Bacillus silvestris and Bacillus pasteurii. Although not displayed by all strains, mosquito pathogenicity is a noteworthy characteristic of B. sphaericus sensu lato. Phylogenetic analysis based on 16S rDNA sequences from 58 strains identified as B. sphaericus was used to examine the genetic heterogeneity of the taxon. Results from sequence analysis were compared with whole-cell fatty acid profiles and other phenotypic determinations. The B. sphaericus-like strains segregated into seven distinct clusters in a phylogenetic tree generated from 16S sequences. One cluster represented B. sphaericus and another B. fusiformis. A third cluster containing all of the pathogenic strains was closely related to, or was possibly part of, the B. fusiformis group. The remaining four groups were distinct and represented unnamed taxa that were more closely related to B. sphaericus and B. fusiformis than to the psychrophilic round-spored species, Bacillus globisporus and Bacillus psychrophilus. Groups based on phenotypic analysis corresponded to the 16S rDNA phylogenetic clusters. Data showed that B. sphaericus was genetically and phenotypically a highly heterogeneous taxon including at least seven genetically distinct taxa. The pathogenic strains were members of a distinct group and not of the species B. sphaericus sensu stricto. This heterogeneity partially accounts for the apparent variability of mosquito pathogenicity among B. sphaericus strains. PMID- 11034480 TI - Methanocalculus pumilus sp. nov., a heavy-metal-tolerant methanogen isolated from a waste-disposal site. AB - A mesophilic hydrogenotrophic methanogen, strain MHT-1T, was isolated from the leachate of a sea-based site for solid waste disposal (the port of Osaka, Japan). Strain MHT-1T was found to be an irregular coccus and was able to use H2/CO2 and formate as energy sources. Acetate was required for growth. The optimum temperature and pH for growth were 35 degrees C and 6.5-7.5, respectively. Strain MHT-1T was resistant to high concentrations of several heavy metals such as CdCl2 and CuSO4. The G+C content of the DNA was 51.9 mol%. Analysis of the 16S rRNA gene revealed that the isolate was a member of the genus Methanocalculus but distinct from its nearest neighbour, Methanocalculus halotolerans, there being a sequence similarity of 98.9%. DNA-DNA hybridization analysis revealed 51% relatedness with the DNA of M. halotolerans strain SEBR 4845T. The optimum NaCl concentration was 1.0%, whereas the optimum in M. halotolerans was 5.0%. A new species, Methanocalculus pumilus, is proposed for strain MHT-1T. The type strain is MHT-1T (= DSM 12632T = JCM 10627T). PMID- 11034481 TI - Subtercola boreus gen. nov., sp. nov. and Subtercola frigoramans sp. nov., two new psychrophilic actinobacteria isolated from boreal groundwater. AB - Psychrophilic actinobacterial isolates from permanently cold groundwater in Finland were characterized using a polyphasic approach. Growth on agar plates was observed at temperatures down to -2 degrees C, with an optimum at 15-17 degrees C, but no growth was observed at 30 degrees C. The peptidoglycan type was B2y and the characteristic diamino acid was diaminobutyric acid. The cell wall sugars of strain K265T were rhamnose, ribose, xylose and mannose and those of strain K300T were glucose, rhamnose and xylose. The polar lipids included phosphatidylglycerol, diphosphatidylglycerol, one unknown phospholipid and two glycolipids. The main whole-cell fatty acids were 12-methyltetradecanoic acid, 14 methylpentadecanoic acid and 14-methylhexadecanoic acid. Large amounts of anteiso 1,1-dimethoxy-pentadecane and also iso-1,1-dimethoxyhexadecane were present as diagnostic markers. The predominant menaquinones were MK-9 and MK-10. The G+C content of the DNA of strains K265T and K300T was 64.4 and 67.8 mol%, respectively. Comparison of 16S rRNA gene sequences revealed that strains K265T and K300T represent a new lineage among the type-B-peptidoglycan actinomycetes. The closest relatives were Clavibacter michiganensis, Frigoribacterium faeni and Rathayibacter rathayi. On the basis of 16S rDNA sequence, G+C content and chemotaxonomical and physiological characteristics, K265T and K300T clearly represent a new genus. The genus Subtercola gen. nov. is described, together with two species, namely Subtercola boreus sp. nov. (type strain K300T = DSM 13056T = CCUG 43135T) and Subtercola frigoramans sp. nov (type strain K265T = DSM 13057T = CCUG 43136T). PMID- 11034482 TI - Aerobic endospore-forming bacteria from geothermal environments in northern Victoria Land, Antarctica, and Candlemas Island, South Sandwich archipelago, with the proposal of Bacillus fumarioli sp. nov. AB - Aerobic endospore-forming bacteria were isolated from soils taken from active fumaroles on Mount Rittmann and Mount Melbourne in northern Victoria Land, Antarctica, and from active and inactive fumaroles on Candlemas Island, South Sandwich archipelago. The Mt Rittmann and Mt Melbourne soils yielded a dominant, moderately thermophilic and acidophilic, aerobic endospore-former growing at pH 5.5 and 50 degrees C, and further strains of the same organism were isolated from a cold, dead fumarole at Clinker Gulch, Candlemas Island. Amplified rDNA restriction analysis, SDS-PAGE and routine phenotypic tests show that the Candlemas Island isolates are not distinguishable from the Mt Rittmann strains, although the two sites are 5600 km apart, and 16S rDNA sequence comparisons and DNA relatedness data support the proposal of a new species, Bacillus fumarioli, the type strain of which is LMG 17489T. PMID- 11034483 TI - Atopobacter phocae gen. nov., sp. nov., a novel bacterium isolated from common seals. AB - Two strains of a Gram-positive, catalase-negative, facultatively anaerobic, rod shaped bacterium isolated from common seals were characterized using phenotypic and molecular taxonomic methods. The two strains closely resembled each other based on their biochemical characteristics, and PAGE analysis of whole-cell protein patterns confirmed their close phenotypic affinity. 16S rRNA gene sequencing showed that the two strains were genetically highly related (99.8% sequence similarity) and that they constitute a new line of descent within the lactic acid group of bacteria. The nearest phylogenetic neighbours of the unknown bacterium were Granulicatella spp., with related taxa such as enterococci, carnobacteria, Desemzia incerta, Lactosphaera pasteurii, Melissococcus plutonius, tetragenococci and vagococci more distantly related. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from seals be classified in a new genus as Atopobacter phocae gen. nov., sp. nov. The type strain of Atopobacter phocae is CCUG 42358T (= CIP 106392T). PMID- 11034484 TI - Phylogenetic analysis of gram-positive bacteria based on grpE, encoded by the dnaK operon. AB - The dnaK operon in gram-positive bacteria includes grpE, dnaJ and, in some members, hrcA as well. Both DnaK and DnaJ have been utilized for constructing phylogenetic relationships among various organisms. Multiple copies exist for dnaK and dnaJ genes in some bacterial genera, as opposed to a single gene copy for grpE and for hrcA, according to the currently available data. Here, we present a partial protein-based phylogenetic tree for gram-positive bacteria, derived by using the amino acid sequence identity of GrpE; the results are compared with the phylogenetic trees generated from 5S rRNA, 16S rRNA, dnaK and dnaJ sequences. Our results indicate three main groupings: two are within low-G+C DNA gram-positive bacteria comprising Bacillus species and Staphylococcus aureus on the one hand and Streptococcus species/Lactococcus lactis/Enterococcus faecalis/Lactobacillus sakei on the other hand; the Mycobacterium species and Streptomyces coelicolor, belonging to the high-G+C DNA gram-positive bacteria, form the third cluster. This hierarchical arrangement is in close agreement with that obtained with 16S rRNA and DnaK sequences but not DnaJ-based phylogeny. PMID- 11034485 TI - Emendation of genus Collinsella and proposal of Collinsella stercoris sp. nov. and Collinsella intestinalis sp. nov. AB - Collinsella aerofaciens-like strains isolated from human faeces were characterized by biochemical tests, cell wall murein analysis and 16S rDNA analysis. The results indicated that these strains are phylogenetically a member of the family Coriobacteriaceae and close to the genus Collinsella. Their phenotypic characters resembled those of Collinsella aerofaciens. Determination of DNA-DNA relatedness showed that these strains could be divided into two groups (groups 1 and 2). Collinsella aerofaciens and both new groups have A4-type cell wall murein. Based on their phenotypic and phylogenetic characters, two new species of the genus Collinsella are proposed for the isolated strains: Collinsella stercoris for group 1 and Collinsella intestinalis for group 2. Species-specific PCR primer sets for these two species were also constructed. Using these primer sets, Collinsella stercoris and Collinsella intestinalis can be identified easily and rapidly. PMID- 11034486 TI - The rickettsial outer-membrane protein A and B genes of Rickettsia australis, the most divergent rickettsia of the spotted fever group. AB - The genes for rickettsial outer-membrane protein A (rOmpA), a distinguishing feature of spotted fever group (SFG) rickettsiae, and rOmpB, a genus-specific protein, were identified and sequenced in Rickettsia australis. The amino acid sequences of domains I, III and IV of the R. australis rOmpA share close homology with those of rOmpA of other SFG rickettsiae, but the repeat region (domain II) is dramatically different from that of other known SFG rOmpA. R. australis rOmpB is more similar to rOmpB of other SFG rickettsiae than to that of typhus group rickettsiae. PMID- 11034487 TI - 'Flexispira rappini' strains represent at least 10 Helicobacter taxa. AB - 'Flexispira rappini' is a provisional name given to gram-negative, microaerophilic, motile, spindle-shaped micro-organisms with spiral periplasmic fibres and bipolar tufts of sheathed flagella. Several investigators, including Kirkbride, Romero, and Archer isolated strains possessing this morphology. Previously, the phylogenetic position of three 'Flexispira rappini' strains was determined by 165 rRNA sequencing, which indicated that flexispira were members of the genus Helicobacter. As more organisms with 'F. rappini' morphology were isolated, it became apparent that there were multiple Helicobacter taxa with this distinctive morphology. The purpose of this study was to examine a collection of 36 'F. rappini' strains from diverse habitats by using 165 rRNA sequence analysis. The strains fell into 10 taxa, each possibly representing a novel Helicobacter species. Two of these flexispira taxa were previously named, by us, Helicobacter bilis and Helicobacter trogontum. Currently, none of the flexispira taxa contains enough phenotypically and genotypically characterized strains to be formally named 'Helicobacter rappinii'. PMID- 11034489 TI - Proposal for the reclassification of Thiobacillus novellus as Starkeya novella gen. nov., comb. nov., in the alpha-subclass of the Proteobacteria. AB - Thiobacillus novellus is a facultatively chemolithoautotrophic and methylotrophic, Gram-negative, rod-shaped sulfur bacterium, shown by 16S rRNA gene sequence analysis to be a member of the alpha-2 subclass of the Proteobacteria. As such, it must be excluded from the genus Thiobacillus, whose species are members of the beta-Proteobacteria. It closest phylogenetic neighbour appears to be Ancylobacter, from which it is distinct morphologically and in some physiological characteristics. It is distinct physiologically and biochemically in a number of diagnostic features from Paracoccus versutus, in the alpha-3 subclass of the Proteobacteria and does not appear to be sufficiently closely related to any other genus of the alpha-Proteobacteria to be reassigned to a known genus. The new genus and species name Starkeya novella is proposed for T. novellus. The type strain is ATCC 8093T (= NCIMB 10456T = NCIMB 9113T = DSM 506T = IAM 12100T = IFO 12443T = CCM 1077T). PMID- 11034488 TI - Lactobacillus kimchii sp. nov., a new species from kimchi. AB - A bacteriocin-producing lactic acid bacterium, which was isolated from the Korean fermented-vegetable food kimchi, was subjected to a polyphasic taxonomic study using phenotypic characterization and phylogenetic and genetic methods. This organism (MT-1077T) has phenotypic properties that are consistent with the description characterizing the genus Lactobacillus. Phylogenetic analysis based on 16S rDNA sequences showed clearly that strain MT-1077T is a member of the genus Lactobacillus. The closest phylogenetic relatives are Lactobacillus alimentarius KCTC 3593T and Lactobacillus farciminis LMG 9200T, with levels of 16S rDNA similarity of 98.4 and 98.2%, respectively. Levels of 16S rDNA similarity between strain MT-1077T and other Lactobacillus species were less than 93.0%. Differences in some phenotypic characteristics and DNA-DNA relatedness data indicated that strain MT-1077T should be distinguished from L. alimentarius KCTC 3593T and L. farciminis LMG 9200T. On the basis of the data presented, it is proposed that strain MT-1077T should be placed in the genus Lactobacillus as a new species, Lactobacillus kimchii sp. nov. The type strain of the new species is strain MT-1077T (= KCTC 8903PT = JCM 10707T). PMID- 11034490 TI - Lactobacillus arizonensis sp. nov., isolated from jojoba meal. AB - Five strains of simmondsin-degrading, lactic-acid-producing bacteria were isolated from fermented jojoba meal. These isolates were facultatively anaerobic, gram-positive, non-motile, non-spore-forming, homofermentative, rod-shaped organisms. They grew singly and in short chains, produced lactic acid but no gas from glucose, and did not exhibit catalase activity. Growth occurred at 15 and 45 degrees C. All strains fermented cellobiose, D-fructose, D-galactose, D-glucose, lactose, maltose, D-mannitol, D-mannose, melibiose, D-ribose, salicin, D sorbitol, sucrose and trehalose. Some strains fermented L-(-)-arabinose and L rhamnose. D-Xylose was not fermented and starch was not hydrolysed. The mean G+C content of the DNA was 48 mol%. Phylogenetic analyses of 16S rDNA established that the isolates were members of the genus Lactobacillus. DNA reassociation of 45% or less was obtained between the new isolates and the reference strains of species with G+C contents of about 48 mol%. The isolates were differentiated from other homofermentative Lactobacillus spp. on the basis of 16S rDNA sequence divergence, DNA relatedness, stereoisomerism of the lactic acid produced, growth temperature and carbohydrate fermentation. The data support the conclusion that these organisms represent strains of a new species, for which the name Lactobacillus arizonensis is proposed. The type strain of L. arizonensis is NRRL B-14768T (= DSM 13273T). PMID- 11034491 TI - Mycobacterium kubicae sp. nov., a slowly growing, scotochromogenic Mycobacterium. AB - A previously uncharacterized, slowly growing, scotochromogenic Mycobacterium species was detected by HPLC analysis of the cell-wall-bound mycolic acids. The mycolic acid pattern standard was shown to be a late-eluting, contiguous peak cluster occurring at approximately 8-9 min. The mycolic acid pattern was noted to be most similar in number of peaks and range of elution to that reported previously for Mycobacterium asiaticum. However, the relative distribution of peaks within the elution range demonstrated a pattern with prominent peaks that started to emerge later than the characteristic M. asiaticum pattern. Standard biochemical identification test results were similar to those of the photochromogenic species M. asiaticum. Comparative 16S rRNA gene sequence analysis confirmed the genetic uniqueness of the strains and demonstrated the unclassified mycobacteria to be in a unique, intermediate position between slow and rapid growers in the phylogenetic tree of Mycobacterium. The name Mycobacterium kubicae sp. nov. is proposed for this taxon. The type strain is CDC 941078T (= ATCC 700732T = CIP 106428T). PMID- 11034492 TI - Mycobacterium elephantis sp. nov., a rapidly growing non-chromogenic Mycobacterium isolated from an elephant. AB - A strain isolated from a lung abscess in an elephant that died from chronic respiratory disease was found to have properties consistent with its classification in the genus Mycobacterium. An almost complete sequence of the 165 rDNA of the strain was determined following the cloning and sequencing of the amplified gene. The sequence was aligned with those available on mycobacteria and phylogenetic trees inferred by using three tree-making algorithms. The organism, which formed a distinct phyletic line within the evolutionary radiation occupied by rapidly growing mycobacteria, was readily distinguished from members of validly described species of rapidly growing mycobacteria on the basis of its mycolic acid pattern and by a number of other phenotypic features, notably its ability to grow at higher temperatures. The type strain is Mycobacterium elephantis DSM 44368T. PMID- 11034493 TI - Janibacter terrae sp. nov., a bacterium isolated from soil around a wastewater treatment plant. AB - A bacterial strain, CS12T, which was isolated from soil around a wastewater treatment plant, was subjected to a polyphasic taxonomic study using phenotypic characterizations and genetic methods. The cell wall of strain CS12T contains meso-diaminopimelic acid as the diamino acid but no arabinose and galactose. The predominant menaquinone is MK-8(H4). Mycolic acids are absent. Strain CS12T has a cellular fatty acid profile containing saturated, unsaturated, branched and 10 methyl fatty acids. The major fatty acids are iso-C16:0, C18:1 omega9c and anteiso-C17:0, The G+C content is 69 mol%. A phylogenetic tree based on 16S rDNA sequences showed that strain CS12T forms an evolutionary lineage within the radiation enclosing the members of the family Intrasporangiaceae and, in particular, a coherent cluster with Janibacter limosus DSM 11140T. The level of 16S rDNA similarity between strain CS12T and J. limosus DSM 11140T is 98.7%. The phenotypic characteristics and DNA-DNA relatedness data indicate that strain CS12T should be distinguished from J. limosus DSM 11140T. Therefore, on the basis of the data presented, a new species of the genus Janibacter, Janibacter terrae, is proposed. The type strain of the new species is strain CS12T (= KCCM 80001T = JCM 10705T). PMID- 11034494 TI - Symbiobacterium thermophilum gen. nov., sp. nov., a symbiotic thermophile that depends on co-culture with a Bacillus strain for growth. AB - A Gram-negative and tryptophanase-positive thermophile, whose growth is dependent on co-culture with an associating Bacillus strain, had been reported and tentatively named Symbiobacterium thermophilum strain T(T). Axenic culture of strain T(T) was recently established by dialysing cultures with the supporting bacterial strains or adding their culture broth. Phylogenetic analysis of strain T(T), based on the 16S rDNA sequence, was conducted for the validation of S. thermophilum. The sequence of strain T(T) was located at the outermost position in the high-G+C Gram-positive group distinctly isolated from any other branches hitherto known. Ten sequences identical to that of strain T(T), and one sequence closely related to it, were identified for the first time from soil and compost samples. The outer membrane of strain T(T) had a three-layered structure, outside the cytoplasmic membrane, which is similar to the S-layer in the cells of members of the Bacillaceae. Chemical analysis of the cells revealed that menaquinone-6 is a major component of the quinone system. According to these results, along with several previous observations (i.e. a G+C DNA content of 65 mol% and the identification of iso-C15:0 and iso-C17:0 acids as major cellular fatty acids), the new taxon Symbiobacterium thermophilum gen. nov., sp. nov. is proposed. The type strain is S. thermophilum strain T(T) (= IAM 14863T). PMID- 11034495 TI - Prochlorococcus marinus Chisholm et al. 1992 subsp. pastoris subsp. nov. strain PCC 9511, the first axenic chlorophyll a2/b2-containing cyanobacterium (Oxyphotobacteria). AB - The formal description of Prochlorococcus marinus Chisholm et al. 1992, 299 was based on the non-axenic nomenclatural type, strain CCMP 1375T. The purification and properties of the axenic strain PCC 9511, derived from the same primary culture (SARG) as the type species, are reported here. Prochlorococcus PCC 9511 differs from the latter in possessing horseshoe-shaped thylakoids, exhibiting a low chlorophyll b2 content and lacking phycoerythrin, but shares these phenotypic properties with Prochlorococcus strain CCMP 1378. This relationship was confirmed by 16S rRNA sequence analyses, which clearly demonstrated that the axenic isolate is not co-identic with the nomenclatural type. Strain PCC 9511 has a low mean DNA base composition (32 mol% G+C) and harbours the smallest genome of all known oxyphotobacteria (genome complexity 1.3 GDa = 2 Mbp). Urea and ammonia are the preferred sources of nitrogen for growth, whereas nitrate is not utilized. Several different organic phosphorus compounds efficiently replace phosphate in the culture medium, indicative of ecto-phosphohydrolase activity. In order to distinguish strain PCC 9511 from the nomenclatural type, a new subspecies is proposed, Prochlorococcus marinus Chisholm et al. 1992 subsp. pastoris subsp. nov. PMID- 11034496 TI - Methylarcula marina gen. nov., sp. nov. and Methylarcula terricola sp. nov.: novel aerobic, moderately halophilic, facultatively methylotrophic bacteria from coastal saline environments. AB - A new genus, Methylarcula, with two new species, Methylarcula marina and Methylarcula terricola, are proposed for strains h1T and h37T of moderately halophilic facultatively methylotrophic bacteria isolated from the coastal saline habitats. These methylobacteria are aerobic, Gram-negative, asporogenous, non motile, colourless rods that multiply by binary fission. Their cellular fatty acids profiles consist primarily of straight-chain unsaturated (C18:1; 70-80%), saturated (C18:0; 14-16%) and cyclopropane (C19:0; 5-6%) acids. The major ubiquinone is Q-10. The dominant phospholipids are phosphatidylethanolamine and phosphatidylcholine. Both strains could use methylamine, some sugars and organic acids as carbon and energy sources. They grew well under optimal conditions (29 35 degrees C, pH 7.5-8.5, 0.5-1.0 M NaCl) and accumulated intracellularly poly beta-hydroxybutyrate and the compatible solute ectoine. The ectoine pool was found to increase upon increasing the external NaCl concentration and accounted for 18% of the dry cellular weight. Both strains oxidized methylamine by the N methylglutamate (N-MG) pathway enzymes (gamma-glutamylmethylamide synthetase/lyase and N-MG synthetase/lyase) to formaldehyde and assimilated it via the icl- serine pathway. The DNA G+C content was 60-4 mol% for Methylarcula marina h1T and 57.1 mol% for Methylarcula terricola h37T. The DNA-DNA hybridization value between strains hl and h37 was 25-30%, although they had a low level of DNA relatedness (5-7%) with the type strains of the serine pathway methylobacteria belonging to the genera Methylobacterium, Aminobacter, Methylorhabdus and Methylopila. A comparative 16S rDNA sequence-based phylogenetic analysis placed the two species of Methylarcula into a separate branch of the alpha-3 subclass of the Proteobacteria. The type strains of the new species are Methylarcula marina h1T (= VKM B-2159T) and Methylarcula terricola h37T (= VKM B-2160T). PMID- 11034497 TI - Description of Cellulophaga algicola sp. nov., isolated from the surfaces of Antarctic algae, and reclassification of Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989 as Cellulophaga uliginosa comb. nov. AB - A group of strains with potent extracellular enzymic activity were isolated from the surfaces of the chain-forming sea-ice diatom Melosira and from an unidentified macrophyte collected from the Eastern Antarctic coastal zone. 16S rDNA sequence analysis indicated that the strains belonged to the genus Cellulophaga and showed greatest similarity to the species Cellulophaga baltica (sequence similarity 97%). Phenotypic characteristics, DNA base composition and DNA-DNA hybridization values clearly separate the Antarctic strains from Cellulophaga baltica and other Cellulophaga species. Thus, the strains form a distinct and novel species and have the proposed name Cellulophaga algicola sp. nov. (type strain IC166T = ACAM 630T). In addition, it was recognized that the species Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989, a species phylogenetically remote from the type species of the genus Cytophaga, possessed 16S rDNA sequences and phenotypic and chemotaxonomic traits similar to those of other Cellulophaga species. Thus, it was proposed that the species Cytophaga uliginosa be renamed as Cellulophaga uliginosa comb. nov. PMID- 11034498 TI - Classification of Proteus vulgaris biogroup 3 with recognition of Proteus hauseri sp. nov., nom. rev. and unnamed Proteus genomospecies 4, 5 and 6. AB - Strains traditionally identified as Proteus vulgaris formed three biogroups. Biogroup 1, characterized by negative reactions for indole production, salicin fermentation and aesculin hydrolysis, is now known as Proteus penneri. Biogroup 2, characterized by positive reactions for indole, salicin and aesculin, was shown by DNA hybridization (hydroxyapatite method) to be a genetic species separate from biogroup 1 and from biogroup 3 which is positive for indole production and negative for salicin and aesculin. In this study, 52 strains were examined, of which 36 strains were Proteus vulgaris biogroup 3, which included the current type strain of the species P. vulgaris (ATCC 29905T), and compared to seven strains of Proteus vulgaris biogroup 2 and nine type strains of other species in the genera Proteus, Providencia and Morganella. By DNA hybridization, these 36 strains were separated into four distinct groups, designated as Proteus genomospecies 3, 4, 5 and 6. DNAs within each separate Proteus genomospecies were 74-99% related to each other in 60 degrees C hybridization reactions with < or = 4.5% divergence between related sequences. Proteus genomospecies 3 contained the former P. vulgaris type strain and one other strain and was negative in reactions for salicin fermentation, aesculin hydrolysis and deoxyribonuclease, unlike the reactions associated with strains considered as typical P. vulgaris which are positive in reactions for salicin, aesculin and DNase. Genomospecies 3 can be distinguished from Proteus genomospecies 4, 5 and 6 because it is negative for Jordan's tartrate. Proteus genomospecies 4, containing five strains, was differentiated from Proteus penneri, genomospecies 3 and 6 and most, but not all, strains of genomospecies 5, by its ability to ferment L-rhamnose. Proteus genomospecies 5 and 6, containing 18 and 11 strains, respectively, could not be separated from each other by traditional biochemical tests, by carbon source utilization tests or SDS-PAGE of whole-cell proteins. In an earlier publication, a request was made to the Judicial Commission that the former type strain of P. vulgaris (ATCC 13315) be replaced by P. vulgaris biogroup 2 strain ATCC 29905T, a strain considered more biochemically typical of P. vulgaris strains. This would have the effect of assigning the name P. vulgaris to P. vulgaris biogroup 2. Since this request has been acceded to, the name Proteus hauseri is herein proposed for Proteus vulgaris genomospecies 3. Its type strain is ATCC 700826T. Proteus genomospecies 4, 5 and 6 will remain unnamed until better phenotypic differentiation can be accomplished. All Proteus genomospecies were similar in their antimicrobial susceptibility patterns. Nineteen strains were isolated from urine, four from faeces, two from wounds, nine from other human sources and two from animals. PMID- 11034499 TI - Systematic relationships and cospeciation of bacterial endosymbionts and their carpenter ant host species: proposal of the new taxon Candidatus Blochmannia gen. nov. AB - The systematic relationships of intracellular bacteria of 13 Camponotus species (carpenter ants) from America and Europe were compared to those of their hosts. Phylogenetic trees of the bacteria and the ants were based on 16S rDNA (rrs) gene sequences and mitochondrial cytochrome oxidase subunit I (COI) gene sequences, respectively. The bacterial endosymbionts of Camponotus spp. form a distinct lineage in the y-subclass of the Proteobacteria. The taxa most closely related to these bacteria are endosymbionts of aphids and the tsetse fly. The bacterial and host phylogenies deduced from the sequence data show a high degree of congruence, providing significant evidence for cospeciation of the bacteria and the ants and a maternal transmission route of the symbionts. The cloned rrs genes of the endosymbionts contain putative intervening sequences (IVSs) with a much lower G+C content than the mean of the respective rrs genes. By in situ hybridization specific 16S rDNA oligonucleotide probes verified the presence of the bacteria within tissues of three of the eukaryotic hosts. It is proposed that the endosymbionts of these three carpenter ants be assigned to a new taxon 'Candidatus Blochmannia gen. nov.' with the symbionts of the individual ants being species named according to their host, 'Candidatus Blochmannia floridanus sp. nov.', 'Candidatus Blochmannia herculeanus sp. nov.' and 'Candidatus Blochmannia rufipes sp. nov.'. PMID- 11034500 TI - Sinorhizobium meliloti associated with Medicago sativa and Melilotus spp. in arid saline soils in Xinjiang, China. AB - Of 42 rhizobial isolates from Medicago sativa and Melilotus spp. growing in arid saline fields in Xinjiang, China, 40 were identified as Sinorhizobium meliloti by a polyphasic approach. However, diverse groups were obtained from these isolates in numerical taxonomy and SDS-PAGE of proteins. They could grow at pH 10.5 and were tolerant to 2.5-4.0% (w/v) NaCl. PMID- 11034501 TI - High-resolution phylogenetic analysis of NO2--oxidizing Nitrobacter species using the rrs-rrl IGS sequence and rrl genes. AB - A high-resolution phylogenetic analysis of Nitrobacter strains and their neighbours was made using the rrs-rrl intergenic spacer sequence and the hypervariable part of the rrl gene. The phylogenetic tree obtained was consistent with that which was obtained previously but was much more discriminating, permitting the design of genus-specific primers. PMID- 11034503 TI - DNA-DNA relatedness among Thermoactinomyces species: Thermoactinomyces candidus as a synonym of Thermoactinomyces vulgaris and Thermoactinomyces thalpophilus as a synonym of Thermoactinomyces sacchari. AB - DNA-DNA relatedness of all validly described Thermoactinomyces species was determined to infer the genetic relationships between them. The levels of DNA-DNA relatedness among the type strains of Thermoactinomyces species ranged from 2-5 to 92.8%. Based on DNA relatedness data, the type strains of Thermoactinomyces intermedius, Thermoactinomyces putidus, Thermoactinomyces dichotomicus and Thermoactinomyces peptonophilus were considered to be distinct species of the genus Thermoactinomyces. However, the relationship between the type strains of Thermoactinomyces vulgaris and Thermoactinomyces candidus and the relationship between the type strains of Thermoactinomyces sacchari and Thermoactinomyces thalpophilus were reevaluated from levels of DNA-DNA relatedness. The independent DNA relatedness values between Thermoactinomyces vulgaris KCTC 9076T and Thermoactinomyces candidus KCTC 9557T were 90.8 and 92.8%. Thermoactinomyces thalpophilus KCTC 9789T and Thermoactinomyces sacchari KCTC 9790T exhibited independent values of 85.6 and 87.3%. Accordingly, on the basis of DNA-DNA relatedness data together with 16S rDNA sequence data determined recently, it is proposed that Thermoactinomyces candidus should be considered as a synonym of Thermoactinomyces vulgaris and Thermoactinomyces thalpophilus be considered as a synonym of Thermoactinomyces sacchari. PMID- 11034502 TI - Janibacter brevis sp. nov., a new trichloroethylene-degrading bacterium isolated from polluted environments. AB - Two strains that were strongly able to degrade trichloroethylene were isolated from contaminated environmental samples. The isolates are non-motile, aerobic, non-spore-forming, gram-positive coccoid to short rods with cell-wall peptidoglycan containing meso-diaminopimelic acid. The major type of menaquinone is MK-8(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol. Mycolic acids were absent. The G+C content of the DNA was 72 mol%. Furthermore, comparison of 16S rDNA sequences shows that the isolates clearly belong to the genus Janibacter. The isolates differ from Janibacter limosus at the species level, as shown by the value for DNA-DNA hybridization. It is therefore proposed that the strains be assigned to the genus Janibacter as Janibacter brevis sp. nov. PMID- 11034504 TI - Nocardiopsis kunsanensis sp. nov., a moderately halophilic actinomycete isolated from a saltern. AB - A moderately halophilic actinomycete, designated HA-9T, was isolated from a saltern in Kunsan, Republic of Korea, and was the subject of polyphasic identification. Analysis of 16S rDNA indicated that the isolate belonged to the genus Nocardiopsis, but differed genetically from other Nocardiopsis species. Strain HA-9T contained meso-diaminopimelic acid, no diagnostic sugars, hexa- or octa-hydrogenated menaquinones with 10 isoprene units, straight-chain saturated or monounsaturated, iso-, anteiso-, 10-methyl branched fatty acids with 13-18 carbons and type III phospholipids. All of these characters consistently assign the isolate to the genus Nocardiopsis. All of the validly described Nocardiopsis species, including moderately halophilic Nocardiopsis halophila, can be differentiated from the saltern isolate using morphological and physiological traits. On the basis of polyphasic evidence, the name Nocardiopsis kunsanensis sp. nov. is proposed for strain HA-9T (= KCTC 9831T), which is designated the type strain. PMID- 11034505 TI - Leuconostoc kimchii sp. nov., a new species from kimchi. AB - A Gram-positive, catalase-negative, facultatively anaerobic, coccus-shaped bacterium, designated IH25T, was isolated from kimchi, a traditional Korean vegetable product. Phylogenetic analysis based on almost complete 16S rDNA sequences placed the isolate in a monophyletic clade corresponding to the genus Leuconostoc. All validly described species in the genus Leuconostoc, with the exception of Leuconostoc fallax, showed high sequence identity of over 97%. The 16S rDNA sequence of strain IH25T showed the highest homology to those of Leuconostoc gelidum DSM 5578T (98.9%) and Leuconostoc citreum KCTC 3526T (98.3 %). However, DNA-DNA hybridization experiments indicated that the organism represents a novel genomic species in the genus, since the previously known leuconostocs share DNA homology with strain IH25T of less than 70%. In this work, it is proposed that isolate IH25T be classified in the genus Leuconostoc as Leuconostoc kimchii sp. nov. The type strain of Leuconostoc kimchii is IH25T (= KCTC 2386T = IMSNU 11154T). PMID- 11034506 TI - Molecular genotyping of human Ureaplasma species based on multiple-banded antigen (MBA) gene sequences. AB - Ureaplasma urealyticum has been divided into 14 serovars. Recently, subdivision of U. urealyticum into two species has been proposed: U. parvum (previously U. urealyticum parvo biovar), comprising four serovars (1, 3, 6, 14) and U. urealyticum (previously U. urealyticum T-960 biovar), 10 serovars (2, 4, 5, 7 13). The multiple-banded antigen (MBA) genes of these species contain both species and serovar/subtype specific sequences. Based on whole sequences of the 5'-ends of MBA genes of U. parvum serovars and partial sequences of the 5'-ends of MBA genes of U. urealyticum serovars, we previously divided each of these species into three MBA genotypes. To further elucidate the relationships between serovars, we sequenced the whole 5'-ends of MBA genes of all 10 U. urealyticum serovars and partial repetitive regions of these genes from all serovars of U. parvum and U. urealyticum. For the first time, all four serovars of U. parvum were clearly differentiated from each other. In addition, the 10 serovars of U. urealyticum were divided into five MBA genotypes, as follows: MBA genotype A comprises serovars 2, 5, 8; MBA genotype B, serovar 10 only; MBA genotype C, serovars 4, 12, 13; MBA genotype D, serovar 9 only; and MBA genotype E comprises serovars 7 and 11. There were no sequence differences between members within each MBA genotype. Further work is required to identify other genes or other regions of the MBA genes that may be used to differentiate U. urealyticum serovars within MBA genotypes A, C and E. A better understanding of the molecular basis of serotype differentiation will help to improve subtyping methods for use in studies of the pathogenesis and epidemiology of these organisms. PMID- 11034507 TI - Three new species in the Saccharomyces sensu stricto complex: Saccharomyces cariocanus, Saccharomyces kudriavzevii and Saccharomyces mikatae. AB - On the basis of genetic analysis, molecular karyotyping and sequence analyses of the 18S rRNA and internal transcribed spacer (ITS) region, three new Saccharomyces species are described, Saccharomyces cariocanus (with type strain NCYC 2890T), Saccharomyces kudriavzevii (with type strain NCYC 2889T) and Saccharomyces mikatae (with type strain NCYC 2888T). Genetic and molecular analyses did not confirm the previously observed conspecificity of Saccharomyces paradoxus and S. cariocanus. The latter species exhibits postzygotic isolation from representative strains from all known geographical populations of S. paradoxus: European, Far-East Asian, North American and Hawaiian. PMID- 11034508 TI - Phylogeny of the bodonid flagellates (Kinetoplastida) based on small-subunit rRNA gene sequences. AB - The phylogeny of kinetoplastid flagellates was investigated by determining the sequences of the small-subunit (18S) rRNA from Bodo designis, Bodo saltans K, Bodo saltans P, Bodo sorokini, Bodo sp. (cf. uncinatus), Cruzella marina, Cryptobia helicis, Dimastigella mimosa and Parabodo nitrophilus and analysing these data together with several previously obtained sequences. The root of the kinetoplastid tree was tentatively determined to be attached to the branch of B. designis and/or Cruzella marina. Within this topology, the suborder Trypanosomatina appears as a late-emerging monophyletic group, while the suborder Bodonina is paraphyletic. Within the bodonid subtree, the branches of parasitic organisms were intermingled with free-living ones, implying multiple transitions to parasitism. The tree indicates that the genera Cryptobia and Bodo are artificial taxa. In addition, the separation of the fish cryptobias and Trypanoplasma borreli as different genera was not supported. PMID- 11034509 TI - Effects of caudal block on cortisol and prolactin responses to postoperative pain in children. AB - We have performed a randomized controlled study in 60 children (ASA 1.8 month to 10 yrs) undergoing lower abdominal and genitourinary surgery, to assess the effects of caudal anaesthesia on plasma cortisol and prolactin concentrations during early postoperative period. After induction of anaesthesia by inhalation or intravenously, thirty children received a standardized general anaesthetic (control group) while the remaining children received caudal anaesthetics with 0.25% bupivacaine in addition to a similar general anaesthetic (caudal group). Blood samples for cortisol and prolactin were taken after induction and 1 hour after surgery. Postoperative analgesia was assessed by modified pain objective scale and side effects were noted. Children in the caudal group had significantly smaller plasma cortisol and prolactin concentrations at 1 hr postoperatively, compared with those in the control group. Plasma cortisol concentration after induction was higher than after one hour of surgery in the caudal group. These results were correlated with pain scores. No serious side effects were recorded. In conclusion, caudal anaesthesia attenuated the postoperative cortisol and prolactin responses to surgery and pain in children. PMID- 11034510 TI - Decrease of cholestasis under "continuous extracorporeal stool transport (CEST)" in prematures and neonates with stomas. AB - According to recent studies, the percentage of neonates suffering a combination of very low birth weight (VLBW) and intestinal malformation or diseases that require surgery has increased. As opposed to mature newborn, split stoma is the therapy of choice in VLBW infants. These neonates and patients with other causes for a congenital or acquired short-bowel syndrome usually require parenteral nutrition and have a high risk of a cholestasis. Until 1993, stoma patients were treated with a special diet in our department, but depending of the stoma site often required parenteral nutrition, while the non-used distal bowel was irrigated with an isotonic electrolyte solution. With this regime, up to 38.5% of the treated patients (10126) were found to have a hyperbilirubinemia (> 3 mg/dl) after the 21st day of life. 1994, we started to use a device that continuously transfers the intestinal chylus from the proximal into the aboral bowel via a pump like a continuous extracorporeal stool transport (CEST). In this study, we evaluated the risk of cholestasis by comparison of the prospectively investigated group (CEST-group 1994-1998) of 33 infants with the results of the retrospectively studied group of 26 patients operated on between 1987 and 1993. We found a significantly lower risk of cholestasis in the CEST-Group (12.9% versus 38.5%; p<0.04). Only 3% of our patients with CEST (one patient) required permanent parenteral nutrition, compared to 50% of the conventionally treated patients. PMID- 11034511 TI - Sodium pertechnetate scintigraphy in detection of Meckel's diverticulum: is it usable? AB - OBJECTIVE: To evaluate the results of 99mTc-Na-pertechnetate scintigraphy in children presenting with symptoms suspicious of Meckel's diverticulum (MD). METHOD: Retrospective study. A total of 55 99mTc-Na-pertechnetate scintigraphies in 53 patients were compared with the results from surgery and other diagnostic procedures and available section reports during the period from 1 Jan. 1981 to 1 Jan. 1996. RESULTS: There were four positive scintigraphies. Three patients underwent a laparotomy and an MD was found. The fourth patient was not operated on and no rebleeding was seen after an observation period of 4 years. A total of 51 scintigraphies were negative. In this group two cases with an MD were found at a later laparotomy for other reasons. We found a sensitivity of 60% on 99mTc-Na pertechnetate scintigraphy in the detection of MD, a specificity and accuracy of 98% and 87%, respectively. CONCLUSION: The 99mTc-Na-pertechnetate scintigraphy has only minor diagnostic value in diagnosing patients suspected of having an MD. Scintigraphic examination should be replaced by diagnostic laparoscopy, which has been reported to be safe and effective and offers the possibility of laparoscopic resection of the MD. PMID- 11034512 TI - An alternative way to cover ventral penile skin defect in Mathieu technique. AB - Mathieu technique is used satisfactorily in distal penile hypospadias without chordee or with minimal chordee. After using this technique, a large defect may sometimes appear on the ventral surface of the penis. To cover the defect, a few techniques, including preputial island flap, Byar's flap and Ombredanne-Nesbit's flap, are used. We describe a new flap to cover the defect more cosmetically. Twenty-four patients ranging in age from one year to 14 years (median age 6.5 years) were operated on. In the patients for whom the defect could not be covered primarily, a longitudinal incision was made along the midline through the penile shaft skin from penile radix up to the border of preputial skin. The relaxed penile skin, which was incised on the dorsal surface, could be approached and sutured easily on the ventral surface without stretching. The new defect that developed on the dorsal surface was closed with the prepuce matching the defect. Nine patients, two with chordee and seven without chordee, underwent this technique. The cosmetic and functional results were excellent in all patients and none of the patient's parents complained about the cosmetic aspect. Only one fistula complication, which healed spontaneously, developed on the 20th postoperative day. Considering these results, we may conclude that excellent cosmetic results can be accomplished by the use of this flap technique. PMID- 11034513 TI - Color Doppler sonography in the diagnosis of acute scrotum in boys. AB - Evaluation of acute painful scrotum is difficult and imaging studies have been reported to be unreliable in detecting testicular torsion. In order to assess the value of color Doppler sonography in acute scrotal disease, the authors reviewed 65 consecutive boys, ranging in age from 1 to 16 years. The study demonstrated absent or diminished flow signals in 11 patients. All of them had a surgical intervention, and in 8 of them, a testicular torsion was confirmed. 54 boys had a symmetrical or increased flow signal on the affected side. Just 5 boys among this group underwent surgical exploration. On follow up, none of the 54 patients with good flow signals proved to have a testicular torsion. Sonography and color Doppler sonography helped to differentiate epididymitis and torsion of a testicular appendage as a basis for further investigations and correct conservative therapy. In detecting a testicular torsion, color Doppler sonography yielded a positive predictive value of 73%, a sensitivity of 100% and a negative predictive value of 100%. We therefore conclude, that Doppler sonography can reliably rule out testicular torsion so that routine scrotal exploration in cases of acute scrotum is no longer necessary. By reducing the number of emergency operations and hospitalization days, color Doppler sonography can cut down the total cost of managing acute painful scrotum in boys. PMID- 11034514 TI - Characteristics of penetrating abdominal firearm injuries in children. AB - Penetrating injuries of the abdomen which occur primarily from gunshots and stabbing become increasingly prevalent in children. Our purpose is to assess the presentation, management and outcome of children with penetrating abdominal firearm injuries (PAFI) and to evaluate the value of Injury Severity Score (ISS) for predicting injury severity and eventual morbidity and mortality in children with PAFI. Between 1985 and 1997, 69 children < or = 15 years of age with PAFI were hospitalized at our institution. Their hospital records were reviewed retrospectively. Of the 69 children with PAFI, 53 were boys and 16 were girls. The mean age was 9.6. The causes of PAFI were explosives (44%), gunshot (39%) and shotgun injuries (17%). The most common organ injured was the small bowel followed by colon and liver respectively. Postoperative complications were seen in 12 children. The mean value of ISS was 16.8; 41 children had an ISS value of > or = 16. The value of ISS was significantly higher in children with complication than in those without complication. The children with ISS value > or = 16 were hospitalized significantly longer than those with < or = 15 of ISS. Particularly shotgun injuries were commonly associated with severe morbidity. ISS can be useful for predicting the severity of injury and the occurrence of morbidity. PMID- 11034515 TI - Congenital anomalies in mice induced by etretinate. AB - We report on the experimental induction of anorectal malformations (ARM) and other internal and external malformations in mouse fetuses induced by maternal administration of etretinate, a long-acting vitamin A analogue. The teratogen was administered to pregnant mice between the 7th and 10th gestational days (E7 and E10). The mice of the control group were given pure sesame oil on E9. We examined survival rates, crown-rump length, and the incidence of internal and external malformations, with particular attention to ARM, in each group. All mice in the E8 group exhibited rectovesical fistula, hydronephrosis, and spina bifida. All males and females in the E9 group given 60 mg/kg of etretinate developed rectoprostatic urethral fistula and rectocloacal fistula, respectively. The E10 group, given 60 mg/kg of etretinate displayed cleft palate (63.6%), forelimb malformation (68.2%), and a short club-shaped tail (100%). The fetuses had more severe types of ARM when etretinate had been administered on an earlier gestational day. The E9 group is a useful model for anorectal malformation, whereas the E8 group is a model for hydronephrosis and spina bifida. PMID- 11034516 TI - Congenital cervical teratoma in neonates. Case report and review. AB - Cervical teratomas are uncommon neoplasms. Although these lesions are histologically benign they are usually large and may cause airway obstruction. Cervical teratomas are usually diagnosed at birth. In-utero diagnosis is possible by prenatal ultrasound which assists in planning early airway management and surgical intervention. Mortality is significant but prognosis is good with airway control and complete surgical excision. However, pressure injury of contiguous structures can limit resectability and adversely affect outcome. Malignant cervical teratoma with metastasis has been reported mostly arising in adults with poor outcome. We present nine cases of neonatal cervical teratoma identified at two institutions between 1984 and 1996. One patient died before surgical intervention. All others underwent resection. There was one intraoperative death and one postoperative death. The remaining six patients did well postoperatively with no significant sequelae with 3 to 14 years follow-up. PMID- 11034517 TI - Choledochocele--a rare cause of necrotising pancreatitis in childhood. AB - Recurring complaints of unknown origin in the upper abdomen during childhood are not frequent. Choledochoceles, or type III choledochal cysts, are rare congenital abnormalities of unclear aetiology. They are rarely considered in the differential diagnosis of upper abdominal symptoms in childhood. Only 130 cases of choledochoceles have been reported in world literature to date. Only 20% of these patients were children or adolescents. The symptoms do not specifically suggest a choledochocele, and are usually attributed to other disorders of the upper intestinal tract, frequently resulting in a delayed diagnosis. We present the case of a 10-year old girl in whom a congenital choledochocele was diagnosed by gastro-intestinal studies and endoscopic retrograde cholangiopancreatography (ERCP) following a 2-years history of recurrent pancreatitis. Various types of choledochoceles are presented in the literature, classified according to morphology and histology. Most authors consider them to be a form of choledochal cyst. Their embryonic development is related to that of congenital duodenal diverticula. Although congenital intraluminal duodenal dilatations are of known oncogenic significance, only two cases of adenocarcinoma arising from a choledochocele have been described to date. Established treatment of large choledochoceles in childhood is transduodenal marsupialization. PMID- 11034518 TI - Laparoscopic removal of a persistent Mullerian duct in a male: case report. AB - The authors refer to the use of the laparoscopic approach in Persistent Mullerian Duct Syndrome, starting from a case of "male vagina" with an anomaly inserted left ductus deferens inside the mullerian persistent duct. This patient was operated on using the laparoscopic technique. PMID- 11034519 TI - Combination of hypospadias and maldescended testis as cardinal symptoms in gonosomal chromosome aberrations. AB - Intersexual genitals or distinct hypospadias in combination with maldescended testis can be caused by endocrinological as well as chromosomal abnormalities. Even in early childhood such clinical findings require specific diagnostic procedures and subsequent treatment which is often invasive but has special importance as regards the early diagnosis of gonadal tumors. We present a child with cryptorchidism on the right, inguinal testis on the left and penoscrotal hypospadias. Cytogenetic analyses revealed a mosaic karyotype 45, X/46, X, idic (Yp) with unequal distribution of the mosaic in different tissues. In consequence of this chromosomal aberration the patient had mixed gonadal dysgenesis which is associated with an increased risk of tumor development in the aberrant gonads. The principles of pediatric, urological, cytogenetic and endocrinological diagnostics and the mode of data collection in the presented case are described and discussed. Furthermore, a protocol for preventive screening is presented, which combines urological and endocrinological investigations in males with malformations of the genito-urinary tract to minimize the risk of tumor development in the aberrant gonads. PMID- 11034520 TI - Laparoscopic cecostomy for anterior ectopic anus with constipation: a new and technical proposal. AB - A female patient of 26 years of age with faecal incontinence is presented. The anal opening was anteriorly located just posterior to the vagina. This had resulted in overflow faecal incontinence due to severe chronic constipation. NMR showed a normal musculature of the pelvic floor and sphincter. Since the patient did not agree to surgery, laparoscopic cecostomy was performed for the first time for antegrade rinsing. PMID- 11034521 TI - Laparoscopic performance of gastrostomy: a simple, safe, and minimal invasive technique. PMID- 11034522 TI - Sexually transmitted diseases among female prisoners in Brazil: prevalence and risk factors. AB - BACKGROUND: Sexually transmitted diseases (STDs) have become an important medical problem in prisons. GOAL: To determine the prevalence of and risk factors for STDs among female inmates in a Brazilian prison. STUDY DESIGN: All female prisoners at the Espirito Santo State Prison were offered enrollment in this cross-sectional study. An interview exploring demographics, criminal charges, and risk behavior was conducted. Blood and genital specimens were collected for STD testing. RESULTS: Of 122 eligible women, 121 (99%) agreed to participate. Prevalence rates were: HIV 9.9%, human T-cell lymphotrophic virus type I 4.1%, hepatitis B virus 7.4%, hepatitis C virus 19%, syphilis 16%, gonorrhea 7.6%, chlamydial infection 11%, human papillomavirus-related cytologic changes 9.3%, trichomoniasis 30%, and bacterial vaginosis 15%. Previous or current drug abuse (54%), injection drug use (11%), and blood transfusion (16%) were associated with at least one STD. Condom use was infrequent. CONCLUSION: The prevalence of STDs and of behaviors leading to ongoing transmission are high among female inmates in Vitoria, Brazil, and demonstrate the potential importance of prevention activities targeting this population. PMID- 11034523 TI - "Shopping" for sexually transmitted disease treatment: focus group discussions among lay persons in rural and urban Zambia. AB - BACKGROUND: In Zambia, persons use different types of services when seeking treatment for sexually transmitted diseases (STDs). GOAL: To gain insight into the rationale behind the selection of treatment sources by investigating perceptions of STDs and by identifying STD treatment sources used. STUDY DESIGN: Focus group discussions were held with 57 men and 44 women in one urban and one rural area in Zambia between May 1997 and June 1997. The focus group discussions were audiotaped and analyzed qualitatively. RESULTS: Participants combined traditional and modern treatment. The main reason given was that even when biomedical medicine was effective, STDs could only be totally cured if the patient was cleansed by traditional herbs. Factors influencing health-seeking behavior were the person's perception, how the diagnosis was determined, type and cost of treatment, demand for sexual partners, and attitudes of health workers, parents, and the church. CONCLUSION: Barriers and enabling factors for the use of treatment sources were identified. Interventions to improve services and cooperation between the health sectors need to consider these factors. PMID- 11034524 TI - Disappointing performance of literature-derived selective screening criteria for asymptomatic Chlamydia trachomatis infection in an inner-city population. AB - BACKGROUND: In an inner-city population with a low prevalence of Chlamydia trachomatis infection, selective screening may be indicated to increase the efficiency of screening. GOAL: To evaluate the performance of sets of selective screening criteria for asymptomatic Chlamydia trachomatis infection in an inner city population. The criteria were derived from reports of studies carried out in various settings. STUDY DESIGN: A total of 5714 women age 15 to 40 years living in Amsterdam were invited for a screening based on home-obtained urine specimens. Criteria identified from the literature were applied to the screening population. A calculated area under the receiver-operator characteristic curve (AUC) of greater than 0.75 was considered a good measure of diagnostic accuracy. RESULTS: Of the four sets of criteria, selection based on the following determinants showed the highest diagnostic accuracy: younger than 25 years, being unmarried, number of partners during the previous 6 months, Surinam or Antillean origin (black), and vaginal douching (AUC, 0.67; 95% CI, 0.65-0.69). Selection based on age alone showed an AUC of 0.57 (95% CI, 0.55-0.69). CONCLUSION: The performance of selective screening criteria for asymptomatic C trachomatis infection in an inner-city population in Amsterdam was insufficient to recommend its implementation in practice. PMID- 11034525 TI - Effectiveness and cost-benefit of enhancements to a syphilis screening and treatment program at a county jail. AB - BACKGROUND: High rates of syphilis are found in inmates of county jails. Treatment of this infected transient population necessitated the development of a rapid protocol. GOAL: To evaluate a rapid screening and treatment protocol for syphilis in a county jail. STUDY DESIGN: Over a 2-year period 18,442 inmates were screened for syphilis with a nontreponemal test and record search for treatment history. Confirmatory test results were reviewed following treatment. Cost was defined as deflated marginal outlays. Benefit was calculated as the discounted expected cost of treatment of congenital, late, and neurosyphilis. RESULTS: The sensitivity, specificity, and positive predictive value of the protocol were 99.6%, 80.8%, and 79.3%, respectively. Of 257 confirmed cases, 183 were offered treatment in jail. The percentage of short-term inmates treated increased following implementation. The cost-benefit ratio was 9.14:1. CONCLUSIONS: The protocol was highly effective in patient identification and treatment delivery, and cost-effective as well. PMID- 11034526 TI - Cost-effectiveness of screening programs for Chlamydia trachomatis: a population based dynamic approach. AB - BACKGROUND: Models commonly used for the economic assessment of chlamydial screening programs do not consider population effects. GOAL: To develop a novel dynamic approach for the economic evaluation of chlamydial prevention measures and to determine the cost-effectiveness of a general practitioner-based screening program. STUDY DESIGN: The dynamic approach was used to estimate the cost effectiveness of a screening program for the first 10 years of screening in The Netherlands. Screening involved a ligase chain reaction test on urine followed by standardized therapies and partner referral. Eligible persons were sexually active, 15 to 24 years, visited a general practitioner, and had no symptoms of sexually transmitted diseases. The heterosexual model population, which consisted of persons 15 to 64 years, had a total chlamydial prevalence of 4.1% before screening. Screening effects on chlamydial incidence were computed by using a population-based stochastic simulation model. Incidence data were connected with a decision analysis model to determine the health effects of the program. The net costs of the program were calculated (investment costs minus averted costs, in 1997 US$) from a societal perspective and expressed per major outcome averted (symptomatic pelvic inflammatory disease, chronic pelvic pain, ectopic pregnancy, infertility, neonatal pneumonia). RESULTS: Unlike the standard static approach, our model adequately addresses dynamic processes such as chlamydial transmission and the impact of screening programs on chlamydial incidence in the population. During the first 10 years of screening, the investigated program yields savings of US $492 or US $1,086 per major outcome averted, excluding or including indirect costs, respectively. These results depend on chlamydial prevalence and partner referral. CONCLUSIONS: The cost-effectiveness of chlamydial screening programs seems best to be determined by using dynamic modeling on a population basis. In addition to preventing negative health outcomes, the investigated screening program may save costs. PMID- 11034527 TI - Clinical utility of measuring white blood cells on vaginal wet mount and endocervical gram stain for the prediction of chlamydial and gonococcal infections. AB - BACKGROUND: White blood cells on endocervical Gram stain and vaginal wet mount are frequently used to predict chlamydial and gonococcal infections. Previous studies provide conflicting evidence for the clinical utility of these tests. GOAL: To evaluate the clinical utility of measuring white blood cells on vaginal wet mount and endocervical Gram stain for the prediction of chlamydial infection and gonorrhea. STUDY DESIGN: Women undergoing pelvic examinations at 10 county health department family planning and sexually transmitted disease clinics were tested for chlamydial infection by ligase chain reaction assay (n = 4550) and for gonorrhea by culture (n = 4402). Vaginal wet mount and endocervical Gram stains were performed in county laboratories at the time of examination. RESULTS: The prevalences of chlamydial infection and gonorrhea were 8.8% and 3.2%, respectively. For detection of chlamydial or gonococcal infection, the likelihood ratio was 2.85 (95% CI, 2.10-3.87) for > 30 white blood cells on vaginal wet mount and 2.91 (95% CI, 2.07-4.09) for > 30 white blood cells on endocervical Gram stain. Similar results were seen for individual diagnoses either of chlamydial infection or of gonorrhea. CONCLUSION: Vaginal wet mount and endocervical Gram stain white blood cells are useful for the presumptive diagnosis of chlamydial infection or gonorrhea only in settings with a relatively high prevalence of infection or when other predictors can increase the likelihood of infection. PMID- 11034528 TI - Hepatitis B prevalence in young women living in low-income areas: the population based San Francisco Bay area's Young Women's Survey. AB - BACKGROUND: The risk of sexually transmitted hepatitis B virus infection is proportionally higher for young adults and women. Low socioeconomic groups have high rates of hepatitis B infection with no identified source of transmission. The prevalence and correlates of transmission of hepatitis B virus among young women of low socioeconomic status have not been well documented. GOAL: To determine the population-based prevalence and correlates of sexually acquired hepatitis B virus infection in young low-income women in the San Francisco Bay Area. STUDY DESIGN: A three-county, door-to-door serosurvey of hepatitis B virus core antibody among young women living in low-income areas was conducted from April 1996 to January 1998. Multivariate analysis of sexually acquired hepatitis B virus infection excluded participants of Asian or Pacific Islander ethnicity or with a history of intravenous drug use or transfusion. RESULTS: The prevalence of sexually acquired hepatitis B virus infection was 6.4% (95% CI, 4.7%-8.1%). Correlates of infection were black race (odds ratio, 3.9; 95% CI, 1.2-11.9 compared with white race) and herpes simplex virus type 2 infection (odds ratio, 2.0; 95% CI, 1.0-3.9). CONCLUSIONS: Young black women have a higher risk of sexually acquired hepatitis B virus infection. Herpes simplex virus type 2 infection may predispose to hepatitis B virus infection and/or be a marker for lifetime sexual risk behavior. PMID- 11034529 TI - Soliciting sex on the Internet: what are the risks for sexually transmitted diseases and HIV? AB - BACKGROUND: Strategies to meet sex partners have been augmented by the Internet. This medium is an environment of potential risk for acquiring or transmitting sexually transmitted disease (STD). GOAL: To document how the Internet is used to find sex partners and what risks such activity poses for STD infection. STUDY DESIGN: Participant observations of 175 chat rooms targeting men who have sex with men (MSM), heterosexuals, and couples seeking sex partners. RESULTS: Findings indicate evidence of past meetings (9% of MSM-room observations, 15% of couple-room observations) and solicitation of sex (9% of heterosexual-room observations, 17% of MSM-room observations, 36% of couple-room observations) by members of these groups. Safer sex or risk-reduction behaviors were not frequently mentioned, but were sometimes acknowledged through solicitation of drug-free and disease-free partners. CONCLUSIONS: Because people can use the Internet to solicit sex partners, it is a risk environment for sexually transmitted diseases. The Internet offers fast and efficient encounters resulting in sexual contact, which may translate into more efficient disease transmission. However, the Internet also offers many possibilities for innovative technologic approaches to promote STD and HIV prevention. PMID- 11034530 TI - Hereditary breast cancer: high risk genes, genetic testing and clinical implications. AB - About one in eight to ten women living in Western countries will develop breast cancer during her lifetime and between 5-10% of these cases result from an inherited susceptibility to the disease. Within the past few years, a number of genes associated with a high risk of breast cancer have been identified, including BRCA1, BRCA2, TP53, PTEN, MLH1, MSH2, and STK11. The identification of these genes, together with the rapid advances in molecular genetic analyses, should improve the diagnosis and therapy of breast cancer. This article reviews the genetic basis of hereditary breast cancer, in particular the contribution of BRCA1 and BRCA2 and discusses the clinical application of this new molecular knowledge with regard to molecular testing, surveillance and prevention in women with a hereditary predisposition to breast cancer. PMID- 11034531 TI - Evaluation of an automated hemostasis testing analyzer, the Trombolyzer Combi. AB - We evaluated the Trombolyzer Combi (Behnk Elektronik, Norderstedt, Germany), an automated hemostasis analyzer, in a clinical setting. Determination of prothrombin time (PT), activated partial prothrombin time (APTT), fibrinogen (FIB) and antithrombin (AT) were performed using Organon Teknika reagents. Determination of PT, APTT and FIB on a KC4 (Amelung, Germany) using Dade reagent (Dade Behring, The Netherlands) and determination of AT on a Hitachi 912 using Chromogenix reagent (Nodia, The Netherlands) were used as reference methods. Within-run and total precision of the tests were determined by measuring pooled plasma samples at various levels in duplicate twice daily for twenty days. For all tests the within-run and total precision of the Trombolyzer Combi was comparable or superior to the reference methods. Methods comparison was performed with 100 patient samples for PT, APTT and FIB and with 50 patient samples for AT. The correlation coefficients between the Trombolyzer Combi values and the results from the reference methods were between 0.87 and 0.98. No effect of hemolysis on the determination of the studied parameters was detected. However, bilirubinemia above 260 micromol/L and triglycerides above 9 mmol/L resulted in erroneous test results. In conclusion, it is shown that the Trombolyzer Combi performs equivalently or better than the reference methods and can be used as a state-of the-art hemostasis analyzer in a clinical laboratory. PMID- 11034532 TI - Long term follow up of the utility of troponin T to assess cardiac risk in stable chronic hemodialysis patients. AB - Thirty long-term, stable hemodialysis patients were followed 24 months to identify any predictable relationship between elevated serum cTnT values and the diagnosis of coronary artery disease and/or the occurrence of a cardiac death. Patients with a baseline cTnT value of >0.1 microg/L were at high risk for life threatening cardiac events during the 2 years follow-up. With regard to predicting a cardiac event, cTnT has a specificity of 93.75% and sensitivity of 81.8% compared to cTnI whose specificity was 87.5% but sensitivity of between 9.1 and 18.2%. CK-MB was the most specific at 100% but had a low sensitivity of 9.1%. The hemodialysis process, while causing an increase in the serum levels of all the markers studied except CK, the increase only proved significant for cTnT. The only markers whose stratification remained consistent over the 2 years where cTnT and CK-MB, for all others a gain or lose was registered. Baseline stratification using cTnT with a cut-off value of >0.1 microg/L offers opportunities to select at risk hemodialysis patients for corrective cardiovascular intervention. PMID- 11034533 TI - Determination of cefditoren (ME 1206) in the plasma of elderly patients with multiple diseases using high-performance liquid chromatography. AB - In the present study we developed a new sensitive and selective HPLC-assay for the determination of cefditoren (ME 1206) in the plasma. The column switching technique was used to regenerate the pre-column and the analytical column separately. This method is suitable for pharmacokinetic studies and for drug monitoring. Thus, we measured plasma samples of 15 elderly patients (age range: 67-88 years), receiving cefditoren-pivoxil therapy because of infections of the respiratory tract. In addition, these patients were treated with several co medications because of further diseases. With our HPLC-method no interfering peaks could be detected at the retention time of cefditoren (10.67 min) or of the internal standard (16.62 min). The lowest plasma concentration of cefditoren to be quantified was 25 ng/ml (R.S.D.= 15.5%, n = 5). PMID- 11034534 TI - Split sample analysis of serum folate levels after 18 days in frozen storage. AB - Reliable measurement of folate is becoming increasingly important as links between dietary folate intake, the use of vitamins containing folic acid, and health outcomes such as birth defects and cardiovascular disease are identified. This study was undertaken to formally assess whether the quantity of folate in serum declines after the serum is frozen and stored. Blood samples from 83 pregnant women were tested for serum folate shortly after collection and again after 18 days of storage at -20 degrees C. A shift from higher to lower serum folate categories was observed after 18 days of storage. For the first test, 40.9 % of the samples were > or = 20 microg/L compared with 19.3 % of the test results on second test. For the 75 samples in the quantifiable range (< 40 microg/L), a mean decrease of 5.0 microg/L (+/- 0.5) of serum folate was observed (p < 0.0001). When compared to serum samples stored in a non frost-free freezer at -20 degrees C or -70 degrees C, serum stored in a frost-free freezer at -20 degrees C for even a short period of time may be relatively unstable and sensitive to minor temperature fluctuations associated with the freeze-thaw cycles. PMID- 11034535 TI - Clinical role of fecal antigen determination in the diagnosis of Helicobacter pylori infection. PMID- 11034536 TI - The importance of antioxidant enzyme and total antioxidant status of patients with acute myocardial infarction on thrombolytic therapy. AB - It is believed that intensive production of free radicals occurs immediately after reperfusion, and that it explains a series of adverse effects of reperfusion. The aim of the study was to establish the importance of the antioxidative enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX) and the total antioxidant status (TAS) in patients with acute myocardial infarction according to success or failure of reperfusion. Enzyme activities in erythrocyte hemolysate and concentration of TAS in plasma were determined by Randox kits in 50 patients with acute myocardial infarction. No significant differences in TAS between the mean values against either clinical status or the 10 time intervals were recorded. The SOD and GPX values were significantly influenced by the presence or absence of reperfusion (p<0.05). Monitoring of SOD and GPX at ten time points: before administration of streptokinase, 1, 3, 6, 12, 18, 24 hours, 2, 3 and 5 days after administration of streptokinase revealed the lowest SOD levels up to one hour before administration of streptokinase, supporting the fact that the maximum production of free radicals is achieved at that time. GPX values differed for 24 hours according to whether reperfusion was achieved or not. The obtained results suggest that free radical production is increased after reperfusion, and monitoring of antioxidant enzymes may help to evaluate the success of thrombolytic therapy. PMID- 11034537 TI - Therapeutic drug monitoring on COBAS INTEGRA 400--evaluation results. AB - The COBAS INTEGRA 400 (Roche Diagnostics GmbH) is a random access analyzer with a consolidated test menue for routine clinical chemistry, specific proteins, drugs of abuse screening and therapeutic drug monitoring (TDM) and different measuring technologies. It was the aim of the present study to evaluate the suitability of this instrument as dedicated analyzer for TDM. Eight assays based on three different technologies were included: Acetaminophen (enzymatic method), Amikacin/ Phenytoin/ Free Phenytoin/ Lidocaine (fluorescence polarization immunoassays; FPIA), Digitoxin/Digoxin (kinetic interaction of microparticles in solution; KIMS). The study comprised the determination of imprecision according to NCCLS EP T protocol, method comparison and linearity studies. The assays were compared with the corresponding methods on AxSYM or TDx analyzers (Abbott Laboratories). For Acetaminophen and Amikacin COBAS INTEGRA 700 was used as additional comparison instrument. The results are summarized in a table (table 6). Precision results are well acceptable with within-run CVs < 5% and total CVs < 6% except for Digitoxin and Digoxin which show a somewhat higher imprecision at low concentrations. Results obtained for Acetaminophen and Amikacin on COBAS INTEGRA 400 and 700 show excellent agreement. A good comparability is also found between COBAS INTEGRA 400 and AxSYM or TDx methods with slight systematic deviations for Acetaminophen, Amikacin and Free Phenytoin. The lower correlation coefficient for the digoxin method comparison can be attributed to two discrepant samples. Linearity throughout the range studied which covered > 80% of the measuring range was confirmed for the five assays tested (Digitoxin, Digoxin, Lidocaine, Free Phenytoin, Phenytoin) based on the acceptance criteria of +/- 10% deviation of the measured values from the theoretical values. Based on the analytical performance of the TDM tests studied it can be concluded that the COBAS INTEGRA 400 is very well suited for routine TDM analysis. PMID- 11034538 TI - Involvement of platelet-derived growth factor in disease: development of specific antagonists. AB - Platelet-derived growth factor (PDGF) is a family of dimeric isoforms that stimulates, e.g., growth, chemotaxis and cell shape changes of various connective tissue cell types and certain other cells. The cellular effects of PDGF isoforms are exerted through binding to two structurally related tyrosine kinase receptors. Ligand binding induces receptor dimerization and autophosphorylation. This enables a number of SH2 domain containing signal transduction molecules to bind to the receptors, thereby initiating various signaling pathways. PDGF isoforms have important roles during the embryonic development, particularly in the formation of connective tissue in various organs. In the adult, PDGF stimulates wound healing. Overactivity of PDGF has been implicated in certain disorders, including fibrotic conditions, atherosclerosis, and malignancies. Different kinds of PDGF antagonists are currently being developed and evaluated in different animal disease models, as well as in clinical trials. PMID- 11034539 TI - Tumor suppression activity of adenovirus E1a protein: anoikis and the epithelial phenotype. AB - Adenovirus E1a proteins reverse-transform diverse human tumor cells in culture. This has stimulated interest in the arenas of clinical and basic cancer research. Clinically, cancer gene therapy trials on E1a are in progress, and drug discovery strategies based on E1a are being considered. Biologically, the effect of E1a is unique in that it overrides most or all oncogenic signaling pathways to yield nontumorigenic cells. Apparently, this is a consequence of the ability of E1a to reprogram transcription in tumor cells so as to produce an epithelial phenotype that is refractory to oncogenic growth stimulation. The molecular basis for this effect is emerging. PMID- 11034540 TI - Comparative analysis of the transforming mechanisms of Epstein-Barr virus, Kaposi's sarcoma-associated herpesvirus, and Herpesvirus saimiri. AB - Members of the gamma herpesvirus family include the lymphocryptoviruses (gamma-1 herpesviruses) and the rhadinoviruses (gamma-2 herpesviruses). Gammaherpesvirinae uniformly establish long-term, latent, reactivatable infection of lymphocytes, and several members of the gamma herpesviruses are associated with lymphoproliferative diseases. Epstein-Barr virus is a lymphocryptovirus, whereas Kaposi sarcoma-associated herpesvirus and Herpesvirus saimiri are members of the rhadinovirus family. Genes encoded by these viruses are involved in a diverse array of cellular signaling pathways. This review attempts to cover our understanding of how viral proteins deregulate cellular signaling pathways that ultimately contribute to the conversion of normal cells to cancerous cells. PMID- 11034541 TI - Genetic predisposition and somatic diversification in tumor development and progression. AB - Studies on human cancer predisposition syndromes have contributed significantly to our understanding on tumor initiation and progression. Work performed on hereditary colon cancer has been particularly fruitful. Much of the molecular background of the various intestinal polyposis syndromes, such as familial adenomatous polyposis (FAP), juvenile polyposis, and Peutz-Jeghers syndrome, has been revealed, pinpointing several key cancer-associated genes. Studies on hereditary nonpolyposis colorectal cancer (HNPCC) have revealed a novel mechanism of tumorigenesis; genomic instability caused by defective DNA mismatch repair (MMR). Understanding the molecular background of these diseases helps us to understand tumor initiation in the affected individuals. Relatively little is known about the details of tumor progression in hereditary and sporadic neoplasia. Certain additional gene mutations can be associated with advancing stages of the disease, but the pace and tempo of the process have remained obscure. A high mutation rate in MMR-deficient tumors has provided a new approach in the analysis of human tumor dynamics. Microsatellite (MS) sequences are frequently mutated in MMR deficient tumors. The high mutation rate allows the use of microsatellite mutations as a tool for analyzing the past patterns of tumor progression. This approach is similar to the use of MS mutations in studying human evolution and migrations. Such tumor studies have revealed progression pathways that differ from the classic adenoma-cancer sequence. The reasons why and how molecular clocks may reveal something new about a well-studied problem are discussed. PMID- 11034542 TI - Primary effusion lymphoma: a liquid phase lymphoma of fluid-filled body cavities. AB - Primary effusion lymphoma (PEL) is a B-cell neoplasm characterized by infection of the tumor clone by human herpesvirus type-8/Kaposi's sarcoma-associated herpesvirus (HHV-8/KSHV) and by liquid growth in fluid-filled body spaces. During its entire clinical course, the lymphoma tends to remain localized to the serous body cavities with no formation of solid tumor masses. The epidemiology of PEL points to a close link with underlying immunodeficiency of the host, as most cases develop in individuals severely immunocompromised because of preexisting acquired immunodeficiency syndrome. The histogenesis and pathogenesis of PEL have been clarified to a sizeable extent by intensive investigations performed since the disease recognition in 1995. PEL is composed of postgerminal center B cells, which bridge immunoblastic and anaplastic features and typically display a non-B, non-T phenotype consistent with late stages of B-cell differentiation. HHV-8/KSHV is thought to play a major role in PEL pathogenesis via expression of several viral latent genes, which have the potential to affect B-cell growth. Other factors involved in PEL pathogenesis include deregulation of cytokine and growth factor autocrine loops, molecular alterations of the tumor DNA, cell cycle abnormalities, stimulation and selection by antigen, and infection by Epstein Barr virus, which occurs in 70% of PEL cases. In the years since the disease discovery, the distinctiveness of the biological and clinicopathological features of PEL has prompted its recognition as an independent lymphoma category by the World Health Organization classification system of hematologic neoplasms. PMID- 11034543 TI - Dimensions of antigen recognition and levels of immunological specificity. AB - Although recognition and specificity are among the most fundamental concepts in immunology, there is a common tendency to equate these notions with the fit, especially in terms of molecular shape, between interacting molecules. Even in the case of monovalent recognition, there are factors that contribute to the energetics of the interaction that are not readily accounted for by detailed structural analysis of the interacting (epitopic and paratopic) molecular surfaces. Consequently, recognition involves more than just the three spatial dimensions and time. Factors such as solute-solvent interactions, molecular crowding, and confinement, not directly related to the details of the intermolecular interface, can play crucial roles in determining both intrinsic affinity and differential intrinsic affinity. Furthermore, stating that a given structural subunit (e.g., amino acid) is recognized in a given noncovalent interaction does not clarify whether the structural subunit in question participates in the interaction through van der Waals contact, contribution to intrinsic affinity, or differential contribution to relative intrinsic affinities for two or more different ligands. Additional factors become relevant in considering the specificity exhibited in multivalent interactions, cell activation, and activation of the whole immune system. Therefore, specificity as defined for a monovalent binding event can diverge from specificity as it is defined for higher-order interactions. A corollary of this conclusion is that the composition of epitopes and paratopes, defined in terms of the structural elements for which substitutions have an effect on the specificity-defining measurement, can differ in different contexts despite complete conservation of the structures that physically make direct contact. An analysis of specificity at the organismal level suggests that the immune system does not recognize or respond to substances that correspond precisely to either nonself substances or to dangerous substances. An alternative notion for the molecular origins of immunological discrimination does not require that there be any single reason for immune responsiveness. This concept of what the immune system recognizes and responds to derives from the recognition that the ultimate function of the immune system is to contribute to survival and reproductive success through any available means. PMID- 11034544 TI - Topoisomerase I-mediated DNA damage. AB - Topoisomerase I is a ubiquitous and essential enzyme in multicellular organisms. It is involved in multiple DNA transactions including DNA replication, transcription, chromosome condensation and decondensation, and probably DNA recombination. Besides its activity of DNA relaxation necessary to eliminate torsional stresses associated with these processes, topoisomerase I may have other functions related to its interaction with other cellular proteins. Topoisomerase I is the target of the novel anticancer drugs, the camptothecins. Recently a broad range of physiological and environmentally-induced DNA modifications have also been shown to poison topoisomerases. This review summarizes the various factors that enhance or suppress top1 cleavage complexes and discusses the significance of such effects. We also review the different mechanisms that have been proposed for the repair of topoisomerase I-mediated DNA lesions. PMID- 11034545 TI - Evidence that cytosine residues within 5'-CCTGG-3' pentanucleotides can be methylated in human DNA independently of the methylating system that modifies 5' CG-3' dinucleotides. AB - In contrast to the complex sequence specificities of the prokaryotic DNA methylating systems, the mammalian machinery identified thus far methylates cytosine residues within the context of a 5'-CG-3' dinucleotide. To explore the possibility that cytosine residues that do not precede guanine may be independently methylated in mammalian DNA, we have examined a region of the human myogenic gene, Myf-3, which is not targeted by the methylating system that methylates 5'-CG-3' dinucleotides. Our investigations have revealed cytosine methylation within the 5'-CCTGG-3' pentanucleotides specified by the 0.8-kb Myf-3 probe. We have also found that in DNA from neoplastic cells, in which 5'-CG-3' dinucleotides within Myf-3 become abnormally hypermethylated, cytosine residues within 5'-CCTGG-3' pentanucleotides are not methylated. Moreover, methylation of 5'-CCTGG-3' pentanucleotides was not detected within the closely related Myf-4 gene, which is normally 5'-CG-3' hypermethylated. These findings indicate the existence of a system that methylates 5'-CCTGG-3' pentanucleotides independently of the system that methylates cytosine residues within 5'-CG-3' dinucleotides. It is possible that the 5'-CCTGG-3' methylating system influences the fate of foreign integrated DNA. PMID- 11034546 TI - Cell fate decisions in a human retinal precursor cell line: basic fibroblast growth factor- and transforming growth factor-alpha-mediated differentiation. AB - The purpose of this study was to determine if immortalized human retinal precursor cells could serve as a model to investigate cues that modulate cell fate and differentiation. We investigated the effects of a variety of growth factors broadly but specifically tested the effects of basic fibroblast growth factor (bFGF) and transforming growth factor (TGF)a in retinal cell differentiation and commitment. To determine the role of exogenously added growth factors in a human retinal precursor cell line (KGLDMSM), established from a first-trimester retina, cells were adapted to grow in a defined medium and exposed to a variety of trophic factors (epidermal growth factor [EGF], neuron growth factor [NGF], TGFalpha, TGFbeta, acidic FGF, and bFGF). Dose-response curves were developed to arrive at optimal concentrations. The neurotrophic potential of growth factors was determined by 3H-thymidine incorporation and bromodeoxyuridine (BrdU) labeling. The identity of the emerging neuronal phenotypes were determined by phase-contrast microscopy, immunolabeling for the neuron-specific antigens neurofilament protein (NF) and neuron-specific enolases (NSE), and photoreceptor-specific antigens (Rho1D4, 7G6) using immunocytochemistry and Western blot analysis. To identify some of the early response genes (c-fos, c-myc) expressed in response to growth factors, Northern blot analysis was performed. Almost all of the factors tested increased the total number of cells with a neuronal phenotype. Potency of growth factors to generate neurons was TGFalpha > bFGF > EGF > NGF. Both TGFalpha and bFGF, alone or in combination, increased the total number of neurons. Most of the neurons generated were photoreceptors, as depicted by the polarized phenotype, expression of photoreceptor-specific antigens, and processes resembling rudimentary outer segments. The increase in photoreceptor-like neurons is possibly attributable to an increase in numbers rather than greater survival. Additionally, the majority of the photoreceptors generated labeled with BrdU and for photoreceptor-specific antigens, suggesting that an inductive effect of bFGF and TGFalpha could occur in the cell cycle or shortly thereafter. Both bFGF and TGFalpha induced the expression of the early response gene c-fos while not altering the expression of c-actin or c-myc. The emergence of a photoreceptor phenotype was confirmed by both immunocytochemistry and Western blot analysis. The immortalized retinal precursor cell line could prove valuable in determining the role of exogenously added growth factors in retinal development and differentiation. Both bFGF and TGFalpha enhance the photoreceptor phenotype in medium-density cultures under conditions of defined medium. The same was confirmed by phase-contrast microscopy, immunocytochemistry, and Western blot analysis. Furthermore, cell fate determination in cultured precursor cells could occur during the late part of the cell cycle or shortly after completion of cell division. The effects of TGFalpha and bFGF seem to be slightly additive. The cell line will be extremely valuable in studying mechanisms of cell commitment and generation of retinal cell types, which could be tested for their potential for transplantation. PMID- 11034547 TI - Paired-like homeodomain proteins Phox2a/Arix and Phox2b/NBPhox have similar genetic organization and independently regulate dopamine beta-hydroxylase gene transcription. AB - The homeodomain transcription factors Arix/Phox2a and NBPhox/Phox2b play a role in the specification of the noradrenergic phenotype of central and peripheral neurons. To better understand the functions of these two factors, we have compared the genetic organization, chromosomal location, and transcriptional regulatory properties of Arix and NBPhox. The gene structure is very similar, with each gene containing three exons and two introns, extending a total of approximately 5 kb. Arix and NBPhox are unlinked in human and mouse genomes. NBPhox is located on human Chromosome 4p12 and mouse Chromosome 5, while Arix is located on human Chromosome 11q13 and mouse Chromosome 7. Both proteins bind to three sites in the promoter proximal region of the rat dopamine beta-hydroxylase gene (DBH). In vitro, Arix and NBPhox form DNA-independent multimers and exhibit cooperative binding to the DB1 regulatory element, which contains two homeodomain recognition sites. Both proteins regulate transcription from the rat DBH promoter, and transcription is synergistically increased in the presence of the protein kinase A catalytic subunit (PKA) plus either Arix or NBPhox. The transcription factors exhibit similar concentration-dependent efficacies, and when they are coexpressed, transcription is stimulated to a value approximately equal to that seen with either factor alone. The N-terminal segment of Arix is essential for transcriptional regulatory activity, and this region bears 50% identity with NBPhox, suggesting a similar mechanism of transcriptional activation of the DBH gene. We conclude from this study that Arix and NBPhox exhibit indistinguishable and independent transcriptional regulatory properties on the DBH promoter. PMID- 11034548 TI - Cloning and promoter analysis of the chicken interferon regulatory factor-3 gene. AB - Interferon regulatory factors (IRFs) are a family of DNA-binding proteins involved in mediating the cellular response to interferons (IFNs) and viral infection. Although extensively studied in mammals, IRFs of other vertebrates have been less well characterized. Previously, we cloned chicken interferon regulatory factor-3 (chIRF-3) mRNA, which is rapidly and transiently induced by double-stranded (ds)RNA. The chIRF-3 mRNA encodes a protein distinct from any known mammalian IRF. Here, we show that chIRF-3 is activated additively by type I and type II IFNs. To delineate the sequence elements required to regulate chIRF-3 expression, we cloned chlRF-3 and 0.48 kb of 5' flanking sequence. Computer analysis of the proximal promoter revealed three putative binding sites for nuclear factor (NF)-kappaB, two overlapping interferon-stimulated response elements (ISREs), and an interferon gamma activating sequence (GAS). The presence of both GAS and ISRE consensus sequences in the chIRF-3 promoter is unique among IRF family members. Both type I and II IFNs, as well as dsRNA and IRF-1, trans activate the promoter in short-term transfection experiments. Mutational analysis of the promoter demonstrated that the putative NF-kappaB binding sites are needed for stimulation by dsRNA but not by either type I or type II IFN and that both the overlapping ISREs and GAS are required for full induction by type I or type II IFN. PMID- 11034549 TI - A novel calcium/calmodulin-regulated kinesin-like protein is highly conserved between monocots and dicots. AB - Recently, a novel kinesin-like protein (KCBP) that is regulated by Ca2+/calmodulin was isolated from dicot plants. A homolog of KCBP has not been reported in monocots. To determine if this motor protein is present in phylogenetically divergent flowering plants, Arabidopsis KCBP cDNA was used as a probe to screen a genomic library of maize, an evolutionarily divergent species. This screening resulted in isolation of a KCBP homolog. Comparison of the predicted amino acid sequence of the KCBP from maize (ZmKCBP), a monocot, with the previously reported KCBP sequences from dicot species showed that the amino acid sequence, domain organization, and gene structure are highly conserved between monocots and dicots. The C-terminal region of ZmKCBP, containing the motor domain and the calmodulin-binding domain, and the N-terminal tail, with a myosin tail homology region (MyTH4) and talin-like region, showed strong sequence similarity to the KCBP homolog from dicots. However, the coiled-coil region is less conserved between monocots and dicots. The ZmKCBP gene contained 22 exons and 21 introns. The location of 19 of the 21 introns of ZmKCBP is also conserved. The ZmKCBP protein is encoded by a single gene and expressed in all tissues. Affinity-purified antibody to the calmodulin-binding domain of Arabidopsis KCBP detected a protein in both the soluble and the microsomal fractions. The C terminal region of ZmKCBP, containing the motor and calmodulin-binding domains, bound calmodulin in the presence of calcium and failed to bind in the presence of EGTA. The ZmKCBP, along with other KCBPs from dicots, was grouped into a distinct group in the C-terminal subfamily of kinesin-like proteins. These data suggest that the KCBP is ubiquitous and highly conserved in all flowering plants and the origin of KCBP predated the divergence of monocots and dicots. PMID- 11034550 TI - A novel chimeric mitochondrial RNA localized in the nucleus of mouse sperm. AB - Six identical cDNA clones corresponding to an RNA of 1685 nucleotides that is enriched in mouse sperm compared with testis were isolated from a mouse testis cDNA library. The sequence of these clones corresponds to the 16S mitochondrial RNA plus an inverted repeat of 120 bp covalently joined to the 5' end of the RNA. By RT-PCR, it was demonstrated that this transcript, referred to as chimeric RNA, was present in mouse sperm, testis, liver, kidney, brain, and spleen. The absence of an equivalent sequence in mitochondrial DNA or as a mitochondrial pseudogene in total DNA extracted from sperm, testis, and somatic tissues suggests that the chimeric RNA is a post-transcriptional product, maybe resulting from a trans splicing reaction. The chimeric RNA was found by RT-PCR in total RNA extracted from purified sperm heads. This result was confirmed by in situ hybridization, which showed clear staining of the sperm nucleus with probes corresponding to sequences of the mitochondrial 16S RNA and the inverted repeat. PMID- 11034551 TI - N-glycosylation contributes to the limited cross-reactivity between hemagglutinin neuraminidase proteins of human parainfluenza virus type 4A and 4B. AB - cDNAs encoding human parainfluenza virus type 4B (hPIV-4B) hemagglutinin neuraminidase (HN) protein were cloned and the nucleotide sequences were determined. A high degree of identity (81.4%) was observed between the nucleotide sequences of hPIV-4A and -4B HN proteins, and an 87.3% identity was found between the deduced amino acid sequences. This degree of identity is considered to be greater than immunological similarity between hPIV-4A and -4B HN proteins determined using monoclonal antibodies. To elucidate the causes of the antigenic difference between HN proteins of hPIV-4A and -4B, we constructed three cDNAs of hPIV-4B HN whose potential N-glycosylation sites were partially or completely the same as in hPIV-4A HN cDNA. We compared the antigenicity of the expressed wild type and mutant proteins, and found that the antigenicities of the mutant hPIV-4B HN proteins were more similar to the hPIV-4A HN protein than to the non-mutant hPIV-4B HN protein. This study indicated that the antigenic diversity between hPIV-4A and -4B was partly caused by deletion or creation of glycosylation sites, showing that the point mutations resulting in deletion or creation of glycosylation sites is one of the initial steps leading to the division of virus into subtypes. PMID- 11034552 TI - GyrA sequence-based typing of Legionella. AB - Comparative sequence analysis of a 423-bp segment of the gyrA gene including a region homologous to the quinolone resistance-determining region (QRDR) of other species was evaluated as a novel typing method for Legionella strains. The study was performed with 29 reference strains representing 11 different Legionella species, with various serogroups, and with 13 clinical isolates of L. pneumophila. Pulsed-field gel electrophoresis and serotyping were employed for comparison of the clinical isolates. QRDR sequencing proved to be a highly discriminative tool for typing Legionellae, and permitted identification of species, serogroups and even different strains within serogroup 1. None of the isolates were resistant to quinolones in vitro and this correlated with dissence of mutations in the QRDR region. The data show that comparative sequence analysis of a short fragment of the gyrA gene is a potentially useful tool for typing of Legionella beyond the serogroup level. It is anticipated that mutations of the QRDR may arise in Legionella as a consequence of the introduction of quinolones as the agents of choice for the treatment of infections with this agent in immunocompromised patients. The employment of QRDR-typing maybe helpful in uncovering such mutations. PMID- 11034553 TI - Enhanced transcription of the s-adenosylhomocysteine hydrolase gene precedes Epstein-Barr virus lytic gene activation in ganglioside-stimulated lymphoma cells. AB - Stimulation of Epstein-Barr virus (EBV) genome-positive Burkitt lymphoma cells with the ganglioside IV3NeuAc-nLcOse4Cer leads to the induction of cell differentiation processes and activates the EBV lytic viral cycle. In cells of the Burkitt lymphoma line Raji differential expression of host cell genes was analysed in the early phase (150 min) post stimulation with the ganglioside to display the cell activities that precede the activation of the EBV lytic cycle using the differential display reverse transcription-polymerase chain reaction technique. Multiple fragment cDNAs derived from control cells and ganglioside stimulated cells were amplified using random primers and displayed via polyacrylamide gel electrophoresis. The expression pattern of 8,400 bands was analysed. Eleven differentially expressed fragment cDNAs were reamplified and identified by nucleotide sequencing. Six of these could be identified as coding for proteins that may take part in virus reactivation and differentiation. The most striking finding was the induction of s-adenosylhomocysteine hydrolase (AHCY) expression. The cellular enzyme AHCY plays an important role in transmethylation reactions controlling the replication of several viruses. Thus. an involvement in EBV replication can be suggested. PMID- 11034554 TI - Frequent contamination of Chlamydia trachomatis and Chlamydia pneumoniae strains with mycoplasma. Biological relevance and selective eradication of mycoplasma from chlamydial cultures with mupirocin. AB - Several strains of Chlamydia trachomatis (CT) and C. pneumoniae (CP) from different sources were screened for mycoplasma contamination using a sensitive nested 16S rDNA polymerase chain reaction-specific for a broad range of mycoplasma species. Five of nine CT and 5/16 CP isolates were contaminated by mycoplasma. Mycoplasma fermentans, M. hyorhinis and M. hominis were found as contaminating agents. To our knowledge no data are available on whether coinfection of chlamydia with mycoplasma alters the biological behavior of chlamydia. Analysis of the biological effect of mycoplasma on chlamydial infection showed a profound mycoplasma-induced reduction of chlamydial growth. Mycoplasma were efficiently eliminated from chlamydial cultures in HEp-2 cells by treatment with mupirocin without affecting chlamydial replication or host cell growth. Two chlamydial strains, C. trachomatis serovar K and one clinical isolate of C. pneumoniae were purged by this method. PMID- 11034555 TI - Incomplete penetrance of MHC susceptibility genes: prospective analysis of polygenic MHC-determined traits. AB - We propose an approach to understanding incomplete penetrance of disease susceptibility genes as a method of studying the underlying mechanisms of polygenic diseases. Incomplete penetrance is the failure of genetically susceptible individuals to exhibit a trait. We define as baseline penetrance that which occurs in genetically identical (monozygotic) twins of an index subject with a major histocompatibility complex (MHC)-associated disease or trait. We consider two mechanisms for incomplete baseline penetrance: an extrinsic (environmental) trigger and an intrinsic stochastic, gene-associated process. The latter can be detected for dominant expression because susceptibility genes in homozygotes (with their two intrinsic triggers) will be up to twice as frequently penetrant as those in heterozygotes. The extent of MHC and non-MHC gene contribution determines differences between baseline penetrance and apparent penetrance in MHC-identical sib pairs, sib pairs in general and MHC-identical unrelated individuals. Inheritance patterns in families do not reveal modes of inheritance of incompletely penetrant polygenic MHC-determined traits. A method is proposed to study such traits prospectively in persons presumed to be homozygous, heterozygous or non-carrying for susceptibility genes by determining trait expression in homozygotes, heterozygotes or non-carriers of trait associated conserved extended MHC haplotypes. The method provides direct estimates of apparent penetrance rates, modes of genetic determination, and, if the trait is dominant, the origin of penetrance. When applied to dominant MHC susceptibility gene-determined immunoglobulin deficiencies in two populations, the ratios of affected haplotype homozygotes to heterozygotes near 2.0 were consistent with an intrinsic mechanism for baseline penetrance acting on the MHC susceptibility genes. PMID- 11034556 TI - Prospective analysis suggests susceptibility genes for deficiencies of IgA and several other immunoglobulins on the [HLA-B8, SC01, DR3] conserved extended haplotype. AB - The extended major histocompatibility complex (MHC) haplotype [HLA-B8, SC01, DR3] is increased in frequency among patients with immunoglobulin (Ig)A deficiency and common variable immunodeficiency. Because the genomic region from HLA-B to HLA DR/DQ is virtually the same on all instances of the haplotype in the general population, we reasoned that all independent instances of [HLA-B8, SC01, DR3] carry MHC susceptibility genes for these disorders. To define immunoglobulin deficiencies determined by genes on this haplotype and their mode of expression and penetrance, serum immunoglobulin class and IgG subclass concentrations were determined prospectively in homozygotes and heterozygotes of this haplotype and in Caucasian controls. Prevalence of individual immunoglobulin deficiencies in persons with [HLA-B8, SC01, DR3] ranged from 13% to 37%, significantly higher than rates in non-carriers or general controls. We found significantly increased frequencies of IgA and IgG4 deficiency only in homozygotes (13.3% and 30%, respectively) compared with heterozygotes (1.7% and 3.4%) or non-carriers (1.6% each), suggesting recessive expression. In contrast, IgD and IgG3 deficiencies were significantly more common in both homozygotes (36.7% and 30%) and heterozygotes (20.3% and 17.5%) compared with controls (4.9% and 3.4%), suggesting dominant inheritance. These results indicate multiple distinct susceptibility genes, some recessive and others dominant, for deficiency of IgA, IgD, IgG3 or IgG4 (but not for IgE, IgG1, IgG2 or IgM) on [HLA-B8, SC01, DR3]. These observations may also help to explain the observed associations of [HLA-B8, SC01, DR3] with both IgA deficiency and common variable immunodeficiency and the common occurrence of IgG subclass deficiencies in some patients with IgA deficiency. PMID- 11034557 TI - HLA-DRB1 and DQB1 genotypes in patients with insulin-dependent neonatal diabetes mellitus. A study of 13 cases. AB - Insulin-dependent neonatal diabetes mellitus (NDM) is a rare form of diabetes with a heterogeneous genetic background. The HLA-DRB1 and DQB1 genotypes were determined for 13 patients with NDM, from 9 unrelated families. Four patients had permanent NDM (PNDM) and 9 patients had transient NDM (TNDM). No excess of HLA susceptibility markers for type 1 diabetes (IDDM) was observed in this series of patients, whatever the forms of diabetes PNDM or TNDM. Paternal isodisomy of chromosome 6 was observed in two TNDM cases. These observations are consistent with the current hypothesis that there is a recessive susceptibility gene, at least in the transient form of the disease, unlinked to the MHC locus on chromosome 6. Although established in a short series, our results do not support an additive role of IDDM1 in the progression of the disease. PMID- 11034558 TI - DLA-DQB1 alleles and bone marrow transplantation experiments in narcoleptic dogs. AB - Human narcolepsy is a neurological disorder known to be tightly associated with HLA-DQB1*0602. A clinically similar disorder has been described in various dog breeds. The canine form of the disease is inherited as an autosomal recessive disorder in Labrador retrievers and Doberman pinschers (canarc-1) but occurs sporadically in other breeds, most typically dachshunds and poodles. In this study, we have examined if there is a relationship between the development of narcolepsy and specific dog leukocyte antigen (DLA)-DQB1 alleles. Ninety-nine dogs were typed for DLA-DQB1-31 with narcolepsy and 68 control animals. Recent studies have linked the development of autosomal recessive canine narcolepsy to a disruption of the hypocretin receptor 2 (Hcrtr2) gene on the same chromosome as the canine MHC region (CFA12), but not close to the DLA. Four Hcrtr2-positive families (two Doberman pinscher families, one Labrador retriever family, one dachshund family) were analyzed at the DLA-DQ level. No relationship was found between narcolepsy and DLA in Hcrtr2-mediated narcolepsy but loose genetic linkage was observed (Zmax=2.3 at theta=25%, m= 40). Bone marrow transplantation between two DLA identical affected (Hcrtr2-/-) and unaffected (Hcrtr2+/-) siblings was also performed and found not to be successful neither in transmitting narcolepsy nor in relieving the symptoms in Doberman pinschers. DLA DQB1 was next studied in 11 dogs with sporadic (non-familial) narcolepsy and in unrelated control animals of the same and different breeds. The allelic and carrier frequencies of various DLA-DQB1 alleles were analyzed. There was no strong positive or negative correlation between the development of narcolepsy and specific DLA-DQB1 alleles. These results do not support the involvement of DLA-DQ in canine narcolepsy, whether of sporadic or familial origin. PMID- 11034559 TI - Homozygous HLA-G*0105N healthy individuals indicate that membrane-anchored HLA-G1 molecule is not necessary for survival. AB - HLA-G is expected to play an important role during fetal development. Recently, a healthy individual homozygous for the HLA-G*0105N allele has been described, suggesting that HLA-G expression was not essential for fetal survival. We now report studies of one family with five healthy siblings homozygous for HLA G*0105N, who had been normally delivered; three of these siblings were females who also had normal deliveries. In addition, HLA-G*0105N cDNA has been fully sequenced, and normal G1 membrane anchored protein cannot be translated since after the codon 130 cytosine deletion (exon 3) a reading frameshift is observed leading to the existence of premature stop codon at position 189 (beginning of exon 4). Other protein isoforms (G2, G3 and G6), all containing the leader peptide and the alpha1 domain, are possible and their messenger mRNAs were found; any of these may undertake the necessary HLA-G functions. Our data show that the membrane anchored HLA-G molecule is not necessary in either mother or fetus for a normal pregnancy and survival. Also, individuals homozygous for HLA-G*0105N are healthy and with no indications of immunodeficiency or autoimmunity. PMID- 11034561 TI - Detection of KIR2DL4 alleles by sequencing and SSCP reveals a common allele with a shortened cytoplasmic tail. AB - Single-stranded conformation polymorphism (SSCP) by capillary electrophoresis was assessed as a screening and typing method for alleles of KIR2DL4. Exon 6 was investigated as this exon was reported to include three polymorphic nucleotides. Exon 6, intron 6 and exon 7 were amplified as a single polymerase chain reaction (PCR) product of 650 bp from genomic DNA. The PCR product was sequenced and analysed by SSCP. Exon 7 was found to be invariant. Only two nucleotides were found to be polymorphic in exon 6 and another three were found in intron 6. Strong linkage disequilibrium was found between the polymorphic nucleotides resulting in the presence of three alleles in a panel of 20 cell lines. Two alleles differed within intron 6 while the third allele differed at two nucleotides in exon 6. All six possible genotypes were distinguishable by SSCP providing information from both the forward and reverse primers was used. Exon 6 of one allele was one nucleotide shorter than that of the other alleles and the resulting frame shift is predicted to produce a truncated cytoplasmic tail due to a premature stop codon four codons into exon 7. SSCP was found to be an efficient method of typing exons 6 and 7 in a panel of 46 bone marrow donors. All three alleles were found to be common and one was in strong linkage disequilibrium with the presence of another KIR sequence KIR3DS1. PMID- 11034560 TI - Homogenous expression of killer cell immunoglobulin-like receptors (KIR) on polyclonal natural killer cells detected by a monoclonal antibody to KIR2D. AB - The activity of human natural killer (NK) cells is in part regulated by the expression of killer cell immunoglobulin (Ig)-like receptors (KIR) that recognize major histocompatibility complex (MHC) class I and can inhibit NK cell cytotoxicity. A monoclonal anti-KIR antibody was established and designated Lig1. Lig1 was shown to be specific for KIR in cell-surface staining and to react with all KIR2D, except KIR2DL4 which lacks a D1 domain, but not with KIR3D molecules in an enzyme-linked immunoadsorbent assay (ELISA) and Western blotting. Unlike other anti-KIR antibodies, Lig1 did not inhibit binding of KIR-Ig-fusion proteins to MHC-class I expressing cells nor did it interfere with KIR-mediated inhibition of NK cell cytotoxicity in a functional assay. Lig1 reacted with all NK cells in polyclonal NK populations from different donors, demonstrating that all NK cells express at least one KIR2D receptor. PMID- 11034562 TI - CDw149 antibodies recognize a clustered subset of CD47 molecules associated with cytoplasmic signaling molecules. AB - One of the recently described antigens broadly expressed on human leukocytes is CDw149, which was defined at the 6th Human Leukocyte Differentiation Antigen (HLDA) Workshop by means of 2 monoclonal antibodies (mAbs). Molecular characterization of this antigen has been lacking. In the present study we demonstrate that these anti-CDw149 mAbs actually recognize a clustered subset of a well-defined membrane protein, CD47, also known as integrin-associated protein (IAP). This clustered subset is present on leukocytes but not erythrocytes. The anti-CDw149 mAbs bind with only low affinity to a monomeric (unclustered) subset of CD47 but with high avidity to the CD47 clusters. A fraction of CD47 is associated with large complexes containing cytoplasmic signaling molecules (Src family kinases and heterotrimeric G-proteins) similar to glycosphingolipid enriched microdomains (GEMs), which may explain the previously described signaling capacity of CD47. The low-affinity anti-CD47 mAbs may be useful tools targeting specific receptor complexes involved in cell activation. Specific reactivity of low-affinity mAbs with clustered subsets of cell surface antigens may more generally explain the nature of poorly defined "activation forms" or activation neoepitopes described previously for several cell surface molecules. PMID- 11034563 TI - On the MICA deleted-MICB null, HLA-B*4801 haplotype. AB - A 100-kb deletion including the MICA gene was recently reported in the HLA-B48 (B*4801)-associated haplotype in Japanese. Interestingly, this MICA deletion is accompanied by a MICB null allele, MICB0107N. In order to further investigate the universality of the apparent tight linkage between these two events, we present data on high-resolution deletion mapping of eight HLA-B48-homozygous individuals. Among these, five carried the MICA deletion linked to MICB0107N, as originally reported. Conversely, the remaining three possessed an intact MICA gene of MICA008 or MICA010 allelic variant associated this time with a putative expressed MICB allele, MICB0102. These results may imply that the expression of both MICA and MICB molecules is indispensable to viability through a yet-to-be understood mutual interaction in immune surveillance. PMID- 11034564 TI - Nucleotide sequence of a new Fc gamma receptor IIIB allele that codes for a neutrophil antigen. AB - A new allele of the human neutrophil antigen (HNA) system (tentatively called NA2M) was discovered and its nucleotide sequence was determined. NA2M differs in a single nucleotide (193G-->A) from FCGR3B*2(NA2), resulting in an amino acid change (54Glu-->Lys). The frequency of the NA2M gene in the Japanese population was estimated to be 0.008. Granulocytes of individuals possessing NA2M reacted with HNA-1b(NA2)-specific monoclonal antibody (TAG2) in the GIFT assay. PMID- 11034565 TI - Serological and nucleotide sequence analysis of HLA-A*6812. AB - We describe a novel HLA-A allele, HLA-A*6812. HLA-typing of a patient was performed by serology and by DNA-based analyses. Cloning and sequencing of exons 2 and 3 revealed that the genotype consisted of a common HLA-A*0101 allele and the novel HLA-A*6812 allele. HLA*6812 is identical to HLA-A*68012 except for a single non-synonymous nucleotide substitution leading to an exchange of a threonine to an isoleucine residue at position 142 of the HLA-class I alpha chain. The allele was also found in the patient's mother. The novel HLA-A68 molecule is recognised by some but not all of our HLA-A28-specific sera. These results confirm that position 142 is important for the serological properties of the HLA-A molecule. PMID- 11034566 TI - Identification of a new HLA-DRB4 allele (DRB4*01033) by PCR-SSP and direct sequencing. AB - In this report, we describe the identification of a novel DRB4*01 allele, DRB4*01033, found in two Spanish Caucasian individuals. The new allele was detected during routine HLA typing by an unusual pattern of amplification obtained by polymerase chain reaction using sequence-specific primers (PCR-SSP) that did not match with any of the previously described DRB4 alleles. In order to establish the polymorphism responsible for this pattern exons 2 and 3 of the DRB4 locus were amplified and directly sequenced. The new DRB4*01 allele is identical to DRB4*0103101 except for a single nucleotide substitution in codon 78 (TAC- >TAT). This nucleotide change does not cause an amino acid change as both triplets code for a tyrosine. PMID- 11034568 TI - Nomenclature for factors of the HLA system, update April/May 2000. WHO Nomenclature Committee for Factors of HLA System. PMID- 11034567 TI - DPB1*8501, a novel DPB1 variant in the US Black population. AB - We describe a new DPB1 allele, DPB1*8501, which was identified by sequencing based typing (SBT) in the UCLA exchange. DPB1*8501 is similar to DPB1*2701 with a difference at position 272, (G to A). This difference leads to an amino-acid change of codon 91 from arginine (CGC) to histidine (CAC). Until now this position has been considered conserved. This substitution is located at the 3' site of exon 2, and may interfere with typing strategies using primers or probes located in this region. PMID- 11034569 TI - Nomenclature for factors of the HLA system, update June 2000. WHO Nomenclature Committee for factor of HLA System. PMID- 11034571 TI - Management of the HIV-HCV coinfected patient. PMID- 11034572 TI - Natural history of early gastric cancer. PMID- 11034570 TI - Functional genomics in gastroenterology. PMID- 11034573 TI - Interferon therapy reduces the risk for hepatocellular carcinoma. PMID- 11034574 TI - Photodynamic therapy for dysplastic Barrett's oesophagus: a prospective, double blind, randomised, placebo controlled trial. AB - BACKGROUND AND AIMS: Photodynamic therapy (PDT) is a treatment in which cell damage is achieved by the action of light on a photosensitizing agent. We have assessed the potential use of PDT in the ablation of Barrett's oesophagus. METHODS: Thirty six patients with dysplastic Barrett's oesophagus receiving acid suppression medication with omeprazole were randomised to receive oral 5 aminolaevulinic acid (ALA) 30 mg/kg or placebo, followed four hours later by laser endoscopy. Follow up endoscopy was performed at one, six, 12, and 24 months. RESULTS: Of 18 patients in the ALA group, a response was seen in 16 (median decrease in area in the treated region 30%; range 0-60%). In the placebo group, a decrease in area of 10% was observed in two patients with no change in 16 (median 0%; range 0-10%; treatment v placebo, p<0.001). No dysplasia was seen in the columnar epithelium within the treatment area of any patient in the PDT group. However, in the placebo group, persistent low grade dysplasia was found in 12 patients (p<0.001). There were no short or long term major side effects. The effects of treatment were maintained for up to 24 months. CONCLUSIONS: This is the first randomised controlled trial of PDT for Barrett's oesophagus. It demonstrates that ALA induced PDT can provide safe and effective ablation of low grade dysplastic epithelium. PMID- 11034575 TI - Natural history of early gastric cancer: a non-concurrent, long term, follow up study. AB - BACKGROUND: Controversy has arisen on the natural history of early gastric cancer (EGC). While some emphasise the effectiveness of early detection in reducing mortality from gastric cancer, others insist that EGC is a pseudo-cancer. AIMS/PATIENTS/METHODS: To elucidate the natural history of EGC, a non-concurrent, long term, follow up study was conducted in 71 patients who were diagnosed endoscopically as having EGC, which was confirmed as cancer on biopsy, but in whom surgical resection was not conducted or delayed by more than six months. RESULTS: The natural course of EGC was observed in 56 cases. Over a period of 6 137 months, 20 remained in the early stage while 36 progressed to the advanced stage. The proportion remaining in the early stage consistently decreased with time. Median duration of those who remained in the early stage was estimated as 44 months. The cumulative five year risk for progressing to the advanced stage was 63.0%. In 38 cases there was no evidence for undergoing surgical resection for gastric cancer. The cumulative five year corrected survival was estimated as 62.8% among those unresected. Hazard rate ratio for gastric cancer mortality was 0.65 (p=0.34) for screening detected versus non-screening detected. Hazard rate ratio for gastric cancer mortality was 0.51, significantly lower for patients whose operations were delayed compared with those unresected. CONCLUSIONS: Although EGC showed a relatively long natural history in general, it progressed to the advanced stage with time and led to death from gastric cancer for the most part if left untreated. PMID- 11034576 TI - Potency and stability of C terminal truncated human epidermal growth factor. AB - INTRODUCTION: Epidermal growth factor (EGF) is normally present as EGF(1-53). A variety of C terminal truncated forms have been used in preliminary trials for treating gastrointestinal injury but their relative potency and stability when used in a clinical setting are unclear. Therefore, we compared the biological activity of recombinant EGF(1-53), EGF(1-52), EGF(1-51), and the C terminal peptides EGF(44-53) and EGF(49-53). METHODS: Purity of forms was confirmed by mass spectrometry. Bioactivity of the different EGF forms was determined using [methyl-(3)H] thymidine incorporation into primary rat hepatocytes and their ability to reduce indomethacin (20 mg/kg subcutaneously)/restraint induced gastric injury in rats. Stability of EGF peptides was determined by serial sampling from a syringe driver system containing EGF/4% albumin in saline. RESULTS: Biological activity assays of EGF(1-53), EGF(1-52), and EGF(1-51) gave almost identical thymidine uptake dose-response curves (maximal responses increasing baseline uptake from 4400 (600) cpm (mean (SEM)) to about 22 000 (2000) cpm when EGF was added at 1. 6 nM). EGF(44-53) and EGF(49-53) did not stimulate (3)H thymidine uptake. Control rats had 47 (4) mm(2) damage/stomach, EGF(1-51), EGF(1-52), and EGF(1-53) at 0.16 and 0.80 nmol/kg/h each reduced gastric injury by about 50% and 80%, respectively (both doses p<0.01 compared with control but no significant difference between the different forms). EGF was stable at room temperature for seven days but biological activity decreased by 35% and 40% at two and three weeks, respectively (both p<0.01). Exposure to light did not affect bioactivity. CONCLUSION: EGF(1-51) and EGF(1-52) are as biologically active as full length EGF(1-53) but the C terminal penta- and decapeptides are ineffective. Clinical trials of EGF can probably use infusion systems for at least 48 hours at room temperature and with exposure to light, without reducing biological efficacy. PMID- 11034577 TI - Development of a novel rapid non-invasive screening test for coeliac disease. AB - BACKGROUND: Coeliac disease is one of the commonest underdiagnosed diseases in general practice. The autoantigen recognised by the sera of patients with coeliac disease has recently been identified as tissue transglutaminase. AIMS: We evaluated a simple non-invasive immunological dot blot assay for coeliac disease, suitable for use by the general physician in the ambulatory setting. The sensitivity and specificity of this dot blot assay based on recognition of recombinant human transglutaminase were compared with those of antiendomysial antibodies and an enzyme linked immunosorbent assay. METHODS: Serum samples were analysed from 64 healthy controls, 58 first degree relatives of coeliacs, 74 diseased controls, and 70 biopsy confirmed untreated patients with coeliac disease. Dot blot assay and enzyme linked immunosorbent assay were performed using recombinant human transglutaminase as antigen. RESULTS: The dot blot assay, which can be performed in 20 minutes, was positive in all 70 untreated coeliacs (sensitivity 100%). Among the three control groups, there were three false positive tests by dot blot (specificity 98%), all belonging to the group of healthy subjects. The antiendomysial antibodies test missed five untreated coeliac patients (sensitivity 93%) and was negative in all three control groups (specificity 100%). The specificity of the immunosorbent assay was 99% for IgA and 98% for IgG, while sensitivity was 93% for IgA, 47% for IgG, and 100% for IgA and IgG combined. CONCLUSIONS: The dot blot assay is highly accurate in detecting untreated subjects with coeliac disease and can be performed in the general physician's medical office during the course of a routine examination. This innovative test is a practical, reliable alternative to both the immunofluorescent based antiendomysial test and immunosorbent assay for detection of transglutaminase antibodies for the diagnosis of coeliac disease. PMID- 11034578 TI - Chemotherapy for cancer causes apoptosis that precedes hypoplasia in crypts of the small intestine in humans. AB - BACKGROUND AND AIMS: The mechanism of gastrointestinal damage (mucositis) induced by cancer chemotherapy remains uncertain. The aims of this study were to define the time course and mechanism of small intestinal damage following chemotherapy in humans. METHODS: Patients receiving chemotherapy underwent upper gastrointestinal endoscopy (a maximum of two per patient) with duodenal biopsy prior to chemotherapy and again at 1, 3, 5, and 16 days after chemotherapy. Tissue was taken for morphometry, disaccharidase assays, electron microscopy, and for assessment of apoptosis using the Tdt mediated dUTP-biotin nick end labelling (TUNEL) method. Villus area, crypt length, and mitotic index were measured by a microdissection technique. RESULTS: Apoptosis increased sevenfold in intestinal crypts at one day, and villus area, crypt length, mitotic count per crypt, and enterocyte height decreased at three days after chemotherapy. Disaccharidase activities remained unchanged. Electron microscopy showed increased open tight junctions of enterocytes at day 3, consistent with more immature cells. All indices improved by 16 days. CONCLUSION: Small intestinal mucositis is associated with apoptosis in crypts that precedes hypoplastic villous atrophy and loss of enterocyte height. PMID- 11034579 TI - Heterogeneity of gastric histology and function in food cobalamin malabsorption: absence of atrophic gastritis and achlorhydria in some patients with severe malabsorption. AB - BACKGROUND: The common but incompletely understood entity of malabsorption of food bound cobalamin is generally presumed to arise from gastritis and/or achlorhydria. AIM: To conduct a systematic comparative examination of gastric histology and function. SUBJECTS: Nineteen volunteers, either healthy or with low cobalamin levels, were prospectively studied without prior knowledge of their absorption or gastric status. METHODS: All subjects underwent prospective assessment of food cobalamin absorption by the egg yolk cobalamin absorption test, endoscopy, histological grading of biopsies from six gastric sites, measurement of gastric secretory function, assay for serum gastrin and antiparietal cell antibodies, and direct tests for Helicobacter pylori infection. RESULTS: The six subjects with severe malabsorption (group I) had worse histological scores overall and lower acid and pepsin secretion than the eight subjects with normal absorption (group III) or the five subjects with mild malabsorption (group II). However, histological findings, and acid and pepsin secretion overlapped considerably between individual subjects in group I and group III. Two distinct subgroups of three subjects each emerged within group I. One subgroup (IA) had severe gastric atrophy and achlorhydria. The other subgroup (IB) had little atrophy and only mild hypochlorhydria; the gastric findings were indistinguishable from those in many subjects with normal absorption. Absorption improved in the two subjects in subgroup IB and in one subject in group II who received antibiotics, along with evidence of clearing of H pylori. None of the subjects in group IA responded to antibiotics. CONCLUSIONS: Food cobalamin malabsorption arises in at least two different gastric settings, one of which involves neither gastric atrophy nor achlorhydria. Malabsorption can respond to antibiotics, but only in some patients. Food cobalamin malabsorption is not always synonymous with atrophic gastritis and achlorhydria, and hypochlorhydria does not always guarantee food cobalamin malabsorption. PMID- 11034580 TI - Bifidobacterium strains from resident infant human gastrointestinal microflora exert antimicrobial activity. AB - BACKGROUND AND AIMS: The gastrointestinal microflora exerts a barrier effect against enteropathogens. The aim of this study was to examine if bifidobacteria, a major species of the human colonic microflora, participates in the barrier effect by developing antimicrobial activity against enterovirulent bacteria. METHODS: Antibacterial activity was examined in vitro against a wide range of Gram negative and Gram positive pathogens. Inhibition of Salmonella typhimurium SL1334 cell association and cell invasion was investigated in vitro using Caco-2 cells. Colonisation of the gastrointestinal tract in vivo by bifidobacteria was examined in axenic C3/He/Oujco mice. Antimicrobial activity was examined in vivo in axenic C3/He/Oujco mice infected by the lethal S typhimurium C5 strain. RESULTS: Fourteen human bifidobacterium strains isolated from infant stools were examined for antimicrobial activity. Two strains (CA1 and F9) expressed antagonistic activity against pathogens in vitro, inhibited cell entry, and killed intracellular S typhimurium SL1344 in Caco-2 cells. An antibacterial component(s) produced by CA1 and F9 was found to be a lipophilic molecule(s) with a molecular weight of less than 3500. In the axenic C3/He/Oujco mice, CA1 and F9 strains colonised the intestinal tract and protected mice against S typhimurium C5 lethal infection. CONCLUSION: Several bifidobacterium strains from resident infant human gastrointestinal microflora exert antimicrobial activity, suggesting that they could participate in the "barrier effect" produced by the indigenous microflora. PMID- 11034582 TI - Gastric emptying: a contributory factor in gastro-oesophageal reflux activity? AB - AIMS: To evaluate the impact of total and proximal stomach emptying on 24 hour and postprandial reflux as well as the number of reflux episodes per hour in relation to the impact of lower oesophageal sphincter (LOS) pressure, and oesophageal contractile and clearance function. METHODS: Seventy one outpatients (37 female, 34 male; age 23-82 years) with symptoms suggestive of both delayed gastric emptying and reflux referred for further investigations participated in the study. Gastric emptying of a semisolid 1168 kJ meal and oesophageal clearance of a water bolus (supine) were recorded scintigraphically, reflux by 24 hour pH monitoring, and oesophageal motility manometrically. RESULTS: Slow proximal but not slow distal or total stomach emptying correlated with increased 24 hour and postprandial acid exposure and increased number of reflux episodes/hour. No relationship was found between total or proximal emptying and LOS resting pressure, oesophageal contraction amplitude, percentage of failed contractions, or clearance. Multiple linear regression analyses showed that slow proximal emptying and low LOS pressure contributed significantly to both 24 hour (p=0.0007 and p=0. 0001) and two hour postprandial acid exposure (p=0.007 and p=0.0001). In contrast, the rate of total emptying contributed to neither 24 hour nor postprandial acid exposure. CONCLUSION: Our data suggest that in contrast with total stomach emptying, the rate of proximal stomach emptying contributes to the extent of 24 hour as well as postprandial acid exposure and the number of reflux episodes/hour. PMID- 11034581 TI - Restricted diet rescues rat enteric motor neurones from age related cell death. AB - BACKGROUND: Alone among autonomic neurones, enteric neurones are known to be vulnerable to age related cell death; over 50% may be lost in aging rodents. A previous study demonstrated unexpectedly that neurones of the myenteric plexus from rats fed a restricted diet appeared not to suffer from extensive cell death in contrast with previous studies of ad libitum fed animals. AIMS: To compare myenteric neurone numbers in the ileum of young and aging male Sprague-Dawley rats fed either ad libitum or a restricted diet. METHODS: Neurones were counted in whole mount preparations of rat ileum stained immunohistochemically for the pan-neuronal marker PGP9.5, for choline acetyltransferase, or for nitric oxide synthase, or with NADH or NADPH histochemistry. RESULTS: Neurone numbers in the rat myenteric plexus were substantially affected by the dietary regimen: ad libitum feeding (50-60 g per day of standard rat chow) resulted in the death of about 50% of myenteric neurones in 24 month Sprague-Dawley rats, while numbers were unchanged when the daily dietary intake was halved between the ages of six and 24 months. Animals fed a double restricted diet (15 g per day) showed no cell loss at 30 months, as well as the predicted increase in longevity. Neurone loss was largely complete by 16 months in ad libitum fed animals. Numbers of cholinergic (possibly motor) neurones, as demonstrated by choline acetyltransferase immunohistochemistry, were substantially reduced in ad libitum fed aging rats but not in animals fed a restricted diet. Loss of cholinergic neurones after ad libitum feeding was confirmed by reduced numbers of neurones of a size range matching that of cholinergic neurones. CONCLUSIONS: Ad libitum feeding of adult rats has adverse effects on the survival of myenteric neurones, neurone loss commencing before 16 months of age. Cholinergic neurones appear to be particularly vulnerable to the effects of diet. Restricting dietary intake from six months of age prevents neurone loss almost entirely up to 30 months of age in these rats. PMID- 11034583 TI - Effects of a serotonin 5-HT(4) receptor antagonist SB-207266 on gastrointestinal motor and sensory function in humans. AB - BACKGROUND: Serotonin 5-HT(4) receptors are located on enteric cholinergic neurones and may regulate peristalsis. 5-HT(4) receptors on primary afferent neurones have been postulated to modulate visceral sensation. While 5-HT(4) agonists are used as prokinetic agents, the physiological role of 5-HT(4) receptors in the human gut is unknown. AIMS: Our aim was to characterise the role of 5-HT(4) receptors in regulating gastrointestinal motor and sensory function in healthy subjects under baseline and stimulated conditions with a 5-HT(4) receptor antagonist. METHODS: Part A compared the effects of placebo to four doses of a 5 HT(4) receptor antagonist (SB-207266) on the cisapride mediated increase in plasma aldosterone (a 5-HT(4) mediated response) and orocaecal transit in 18 subjects. In part B, 52 healthy subjects received placebo, or 0.05, 0.5, or 5 mg of SB-207266 for 10-12 days; gastric, small bowel, and colonic transit were measured by scintigraphy on days 7-9, and fasting and postprandial colonic motor function, compliance, and sensation during distensions were assessed on day 12. RESULTS: Part A: 0.5, 5, and 20 mg doses of SB-207266 had significant and quantitatively similar effects, antagonising the cisapride mediated increase in plasma aldosterone and acceleration of orocaecal transit. Part B: SB-207266 tended to delay colonic transit (geometric centre of isotope at 24 (p=0.06) and 48 hours (p=0.08)), but did not have dose related effects on transit, fasting or postprandial colonic motor activity, compliance, or sensation. CONCLUSION: 5 HT(4) receptors are involved in the regulation of cisapride stimulated orocaecal transit; SB 207266 tends to modulate colonic transit but not sensory functions or compliance in healthy human subjects. PMID- 11034584 TI - Radiation induced cytochrome c release causes loss of rat colonic fluid absorption by damage to crypts and pericryptal myofibroblasts. AB - BACKGROUND: Therapeutic or accidental exposure to radiation commonly causes gastrointestinal disturbances, including diarrhoea. Rats subjected to whole body ionising radiation at a dose of 8 Gy lose their capacity to absorb fluid via the descending colon after four days. After seven days, fluid absorption recovers to control levels. AIMS: To investigate the effect of ionising radiation on colonic permeability together with its effect on mitochondria dependent apoptotic signals and intercellular adhesion molecules. METHODS: Rats were irradiated with doses of 0-12 Gy. Colonic permeability was measured by accumulation of fluorescein isothiocyanate (FITC) dextran in crypt lumens. Changes in levels of cytochrome c, caspase 3, E and OB cadherin, beta-catenin smooth muscle actin, and collagen IV were assessed using immunocytochemistry with confocal microscopy. RESULTS: Cytosolic cytochrome c increased after 8 Gy (t(1/2) 1.4 (0.6) hours) and peaked at approximately six hours. Caspase 3 increased more slowly, particularly in crypt epithelial cells (t(1/2) 57 (14.5) hours). Pericryptal myofibroblasts disintegrated within 24 hours as was evident from loss of OB cadherin and smooth muscle actin. This coincided with increased crypt permeability to dextran. Intercellular adhesion between crypt luminal cells was not lost until day 4 when both beta-catenin and E-cadherin were minimal. The half maximal dose-response for these effects was in the range 2-4 Gy. Recovery of colonic transport was concurrent with recovery of pericryptal smooth muscle actin and OB cadherin. The pan caspase inhibitor Z-Val-Ala-Asp.fluoromethylketone (1 mg/kg per day) had a small effect in conserving the pericryptal sheath myofibroblasts and sheath permeability but had no systemic therapeutic effects. CONCLUSIONS: These data suggest that radiation damage to the colon may be initiated by mitochondrial events. Loss of crypt fluid absorption and increased permeability coincided with decreased intercellular adhesion between crypt epithelial cells and loss of pericryptal sheath barrier function. PMID- 11034585 TI - Stimulation of colorectal cancer cell line growth by ET-1 and its inhibition by ET(A) antagonists. AB - BACKGROUND: The vasoactive peptide endothelin 1 (ET-1) acts via two receptors, endothelin receptors A (ET(A)) and B (ET(B)). ET-1 is overexpressed by human cancers in vivo and in vitro and may be mitogenic for cancer cells. METHOD: To elucidate if ET-1 is a growth regulator the following were investigated in human colorectal cancer cell lines (LIM1215 and HT29): ET-1 production by ELISA; ET receptor expression using radioligand autoradiographic techniques; and responsiveness to ET-1, and to ET(A) and ET(B) antagonism by growth measurements. RESULTS: ET-1 was produced by LIM1215 and HT29 cells (21.3 and 41.7 fmol/ml/10(6) cells (24 hours); 22.6 and 71.7 fmol/ml/10(6) cells (48 hours), respectively). ET(A) and ET(B) receptors were expressed by both cell lines. Addition of ET-1 resulted in a dose dependent increase in cell numbers which was significant at 10(-8)-10(-9) M for LIM1215, with the greatest increase at 10(-8) M (32.7% and 28.4% increase above controls at 48 hours and 72 hours; p<0.05) and at 10(-8)-10( 9) M for HT29, with the greatest increase at 10(-9) M (13.4% and 15.7% increase above controls at 48 hours and 72 hours; p<0.05). ET(A) antagonists BQ123 and BQ610, but not the ET(B) antagonist BQ788, inhibited ET-1 induced proliferation of both LIM1215 and HT29 (p<0.05). CONCLUSION: ET-1 can stimulate the proliferation of colorectal cancer cell lines via the ET(A), but not the ET(B), receptor. PMID- 11034586 TI - Evaluation of global DNA hypomethylation in human colon cancer tissues by immunohistochemistry and image analysis. AB - BACKGROUND: Global hypomethylation of DNA is frequently observed in human tumours. This alteration is detected in early adenomas in colorectal tumorigenesis. Information is currently acquired after extraction of DNA from tissues, digestion with nucleases, and analysis by reverse phase chromatography, or treatment with restriction enzymes followed by gel electrophoresis analysis and Southern hybridisation with radiolabelled probes. AIMS: The purpose of our work was to evaluate the global methylation status of DNA in malignant lesions without loosing the histopathological features of the samples. PATIENTS: The investigation was performed on paired normal-tumour tissues from 13 patients undergoing surgical resection of colorectal adenocarcinomas. METHODS: Antibodies raised against 5-methylcytidine can be used to label methyl rich regions in interphase nuclei. This technique was adapted to the study of paraffin embedded tissues and an immunohistochemical method was developed to assess the global methylation status of individual nuclei while preserving cell morphology and tissue architecture. Computer assisted quantification of the staining intensity was performed on malignant and normal zones of human colon tissues to test the correlation between the immunolabelling signal and the respective histological patterns observed. RESULTS: Qualitative and quantitative differences were observed and measured between the normal and malignant part of each sample. Morphologically altered nuclei displayed densely labelled spots within faintly labelled areas whereas normal nuclei were darker and uniformly stained. Image analysis allowed calculation of the average integrated optical density of the nuclei in both types of tissues, demonstrating a constant and significantly lower intensity for the former type of cells. PMID- 11034587 TI - Safety and efficacy of interferon-ribavirin combination therapy in HCV-HIV coinfected subjects: an early report. AB - BACKGROUND: More severe liver disease together with a poor response rate to alpha interferon argue for the use of more potent anti-hepatitis C virus (HCV) therapies in human immunodeficiency virus (HIV)-HCV coinfected patients, but the efficacy and safety of interferon-ribavirin combination therapy in HIV infected subjects are unknown. AIM: To retrospectively evaluate the efficacy and safety of anti-HCV combination therapy in 21 HCV-HIV coinfected patients receiving antiretroviral therapy, and to access the clinical relevance of in vitro inhibition of phosphorylation by ribavirin of potent inhibitors of HIV-that is, zidovudine, stavudine, and zalcitabine. PATIENTS: Twenty one patients were treated with combined antiretroviral therapy including zidovudine (n=8) or stavudine (n=13) (in association with protease inhibitors in 12). All received ribavirin (1000 or 1200 mg/day) and alpha interferon (3 MU three times/week) for chronic hepatitis C infection. All patients had not responded (n=20) or relapsed (n=1) after a previous six month course of alpha interferon therapy. METHODS: HIV viral load (Monitor test) and CD4 cells count were measured at the beginning and every three months during and after ribavirin plus alpha interferon therapy over a mean period of 11 (1) months. Clinical and biological adverse effects were recorded. RESULTS: There was no significant variation in HIV viral load or CD4 cell counts after three or six months of ribavirin therapy compared with baseline values. Of the 21 subjects, three (14%) had an increase in HIV viral load of more than 0.5 log leading to discontinuation of ribavirin in one. Eleven of 21 (52.4%) had initial negative HCV viraemia at three (n=10) or six (n=1) months but only six were polymerase chain reaction negative at the end of therapy, leading to rates for primary response and breakthrough of 23.8% and 28.5%, respectively. Six months after completion of therapy, three patients relapsed (14. 3%) and three (14.3%) had sustained virological response. Median haemoglobin concentration decreased significantly after three and six months of ribavirin therapy (p= 0.0002 and p=0.0003, respectively) leading to withdrawal of therapy in one patient. CONCLUSIONS: These preliminary results show that: (1) despite in vitro interactions between ribavirin, zidovudine, and stavudine, significant variation in HIV replication does not usually occur in HCV-HIV coinfected patients receiving ribavirin and different antiretroviral regimens, including zidovudine and stavudine; (2) alpha interferon and ribavirin combination therapy induced primary and sustained virological responses in 28.5% and 14.3% of treated subjects (who were previous non-responders to interferon therapy), respectively; (3) anaemia is a frequent adverse event. Such results should be confirmed in larger prospective trials. PMID- 11034588 TI - Anti-hepatitis C virus core IgM antibodies correlate with hepatitis C recurrence and its severity in liver transplant patients. AB - BACKGROUND: The significance of immunoglobulin (Ig) M antibody to hepatitis C virus (HCV) core antigen was studied in 60 patients with HCV infection after orthotopic liver transplantation (OLT) diagnosed by polymerase chain reaction. METHODS: Patients were followed up for a mean of 28 months after transplantation. Sera collected three months before transplantation, and one and 12 months after transplantation were analysed for anti-HCV core IgM (HCV-IgM EIA 2.0 assay). After OLT protocol biopsies, procedures were performed routinely every six months. Semiquantitative histopathological assessment of allograft hepatitis was performed using Knodell's score. The results were correlated with clinical features, liver histology findings, and virological features, such as genotype and viraemic levels assessed by a branched DNA assay. RESULTS: One year after liver transplantation, 29/60 (48%) patients had chronic hepatitis on graft biopsy. The presence of anti-HCV core IgM one month (p=0.004) and 12 months (p=0.003) after OLT was positively correlated with recurrence of chronic hepatitis. The positive predictive value of anti-HCV core IgM detected one month after transplantation was 0.88. A significant relationship was observed between severity of graft disease and presence of anti-HCV core IgM 12 months after transplantation. The mean Knodell score was 8.9 in anti-HCV core IgM positive patients compared with 3.6 in those who were anti-HCV core IgM negative (p=0.001). The presence of IgM anti-HCV did not correlate with serum HCV RNA level or HCV genotype. CONCLUSION: We confirm that the presence of anti-HCV core IgM after OLT is a marker of HCV induced graft damage. The recurrence and severity of HCV hepatitis in patients undergoing OLT for HCV cirrhosis is related to the presence of anti-HCV core IgM after liver transplantation. These findings have diagnostic relevance and confirm that measurement of IgM anti-HCV core may help to better monitor the treatment of HCV recurrence after transplantation. PMID- 11034589 TI - Role of N-acetyltransferase polymorphisms in hepatitis B related hepatocellular carcinoma: impact of smoking on risk. AB - BACKGROUND: Persistent infection with hepatitis B virus (HBV) causes chronic phasic necroinflammation and regenerative proliferation in the liver. The sustained hepatocellular proliferation may render chronic HBV carriers more susceptible to the effects of environmental carcinogens. Aromatic amines are potential hepatocarcinogens in humans. N-acetyltransferase (NAT) is involved in the metabolic activation and detoxification of these compounds. AIMS: To investigate if genetic polymorphisms in N-acetylation are related to hepatocellular carcinoma (HCC) among chronic HBV carriers. METHODS: Genotyping of NAT1 and NAT2 was performed using polymerase chain reaction-restriction fragment length polymorphism on peripheral leucocyte DNA from 151 incident cases of HCC and 211 controls. All subjects were male, and were chronic HBV surface antigen carriers. RESULTS: A significant association between NAT2 genetic polymorphism and HCC was observed among chronic HBV carriers who were smokers but not among those who were non-smokers. For smoking HBV carriers, the odds ratios of developing HCC for those heterozygous and homozygous for the NAT2*4 functional allele compared with those without any copies of the functional allele (reference group) were 2.67 (95% confidence interval 1.15-6.22) and 2.58 (95% confidence interval 1.04-6.43), respectively. The interaction between cigarette smoking and the presence of the NAT2*4 allele just failed to reach statistical significance (p=0.06). No association between NAT1 genotype and HCC was evident overall or within the smoking stratified subgroups. CONCLUSIONS: Our results suggest that NAT2 activity may be particularly critical in smoking related hepatocarcinogenesis among chronic HBV carriers. Our data also indirectly support a role for tobacco smoke derived aromatic amines in the aetiology of HCC. PMID- 11034590 TI - Evidence of a systemic phenomenon for oxidative stress in cholestatic liver disease. AB - BACKGROUND: There is considerable evidence indicating that the severity of hepatic damage in individuals with cholestatic liver disease is causally associated with the extent of intrahepatic oxidative stress. Increased levels or accelerated generation of reactive oxygen species and toxic degradative products of lipid peroxidation have been reported in the plasma of individuals with chronic liver disease and animal models of liver disease. Hence, by virtue of their increased presence in the circulation, it is not unreasonable to suppose that they may account for extrahepatic tissue damage in chronic liver disease. MATERIALS AND METHODS: This hypothesis was tested by determining plasma levels of the ubiquitous antioxidant glutathione (GSH) and lipid peroxides (LP), together with assessment of the extent of lipid peroxidation in the kidney, brain, and heart, in 24 day chronically bile duct ligated (CBDL) rats. The extent of lipid peroxidation in tissues was based on measurement of conjugated dienes, lipid peroxides, and malondialdehyde (MDA) content. Data were compared with identical data collected from unoperated control, pair fed, 24 day bile duct manipulated (sham operated), and pair fed sham operated rats. RESULTS: In CBDL rats, total and reduced plasma GSH levels were almost half those determined in all control rats. Plasma, kidney, and heart LP levels were significantly increased in CBDL rats compared with controls. MDA levels were significantly higher in the kidney, brain, and heart homogenates prepared from CBDL rats compared with MDA content measured in tissue homogenates prepared from the four groups of control rats. CONCLUSIONS: Our data show that experimental cholestatic liver disease is associated with increased lipid peroxidation in the kidney, brain, and heart. Hence we have concluded that the oxidative stress in cholestatic liver disease is a systemic phenomenon probably encompassing all tissues and organs, even those separated by the blood-brain barrier. PMID- 11034591 TI - Co-amoxiclav jaundice: clinical and histological features and HLA class II association. AB - BACKGROUND AND AIMS: Jaundice associated with co-amoxiclav has been increasingly recognised. We aimed to characterise its clinical and histological features and to investigate linkage with human leucocyte antigen class II haplotypes. METHODS: We identified cases in the west of Scotland in the period 1991-1997 and performed polymerase chain reaction amplification and oligonucleotide probing on whole blood. RESULTS: Twenty two cases were identified (10 male, mean age 59.1 years). Jaundice occurred a median of 17 days after drug commencement, with a median peak bilirubin level of 225 micromol/l (range 84-598) and median duration of jaundice 69 days (range 29-150). Two patients had primary biliary cirrhosis and two other patients had persistently abnormal liver biochemistry on follow up. One death occurred in a frail elderly woman despite resolving jaundice. The frequency of jaundice was 1 in 78 209 co-amoxiclav prescriptions. Liver biopsy, available in 12 patients, showed perivenular bilirubinostasis, accompanying reactive ceroid laden macrophages, and portal inflammation with focal injury to interlobular bile ducts. Fourteen of 20 patients had DRB1*1501 compared with 27 of 134 controls (p<2.5 x 10(-6); odds ratio (OR) 9.25; relative risk (RR) 6.43). Of these, seven patients were homozygous for DRB1*1501(p< 10(-8); OR 35.54; RR=8.68) compared with two of 134 controls. All patients with DRB1*1501 had the extended haplotype DRB1*1501-DRB5*0101-DQA1*0102-DQB1*0602. There were no clinical or histological differences between genotypes. CONCLUSIONS: Co-amoxiclav associated hepatotoxicity may have a genetic basis and be delayed, severe, and prolonged, although complete recovery is usual. PMID- 11034592 TI - Frequency of p16(INK4A) alterations and K-ras mutations in intrahepatic cholangiocarcinoma of the liver. AB - BACKGROUND: Inactivation of the tumour suppressor gene p16 (CDKN2/MTS-1/INK4A) and K-ras mutations are among the most frequent genetic alterations in human malignancies. AIMS: To investigate the tumour suppressor gene p16 and its possible association with K-ras mutations in intrahepatic cholangiocarcinomas of the liver. METHODS: The status of p16 was evaluated in 41 cholangiocarcinomas by methylation specific polymerase chain reaction, microsatellite analysis, DNA sequencing, and immunohistochemical staining. K-ras mutations were determined by direct DNA sequencing analyses after microdissection. The results obtained were correlated with histopathological variables and patient survival. RESULTS: Hypermethylation of the 5' CpG island of the p16 gene was found in 34 of 41 (83%) carcinomas. Homozygous deletion at the p16 region was present in two (5%), and loss of heterozygosity (LOH) in eight cases (20%). We failed to detect p16 gene missense mutations. K-ras mutations were found in 22 of 41 (54%) cholangiocarcinomas and in two cases of tumour surrounding non-neoplastic liver tissue. All 22 cancers with K-ras mutations also exhibited methylated p16. We failed to observe a correlation between K-ras or p16 status and histopathological factors or prognosis of patients. CONCLUSION: These data suggest that inactivation of the p16 gene is a frequent event in cholangiocarcinoma. The most common somatic alteration is promotor methylation of the p16 gene which is closely associated with K-ras mutations. We failed to establish p16 or K-ras status as independent prognostic factors in these tumours. PMID- 11034593 TI - Clearance of refractory bile duct stones with extracorporeal shockwave lithotripsy. AB - BACKGROUND: Extracorporeal shockwave lithotripsy (ESWL) has been used since the mid-1980s to fragment bile duct stones which cannot be removed endoscopically. Early machines required general anaesthesia and immersion in a waterbath. AIMS: To investigate the effectiveness of the third generation Storz Modulith SL20 lithotriptor in fragmenting bile duct stones that could not be cleared by mechanical lithotripsy. METHODS: Eighty three patients with retained bile duct stones were treated. All patients received intravenous benzodiazepine sedation and pethidine analgesia. Stones were targeted by fluoroscopy following injection of contrast via a nasobiliary drain or T tube. Residual fragments were cleared at endoscopic retrograde cholangiopancreatography. RESULTS: Complete stone clearance was achieved in 69 (83%) patients and in 18 of 24 patients (75%) who required more than one ESWL treatment. Stone clearance was achieved in all nine patients (100%) with intrahepatic stones and also in nine patients (100%) referred following surgical exploration of the bile duct. Complications included six cases of cholangitis and one perinephric haematoma which resolved spontaneously. CONCLUSION: Using the Storz Modulith, 83% of refractory bile duct calculi were cleared with a low rate of complications. These results confirm that ESWL is an excellent alternative to surgery in those patients in whom endoscopic techniques have failed. PMID- 11034594 TI - Columnar cuff cancer after restorative proctocolectomy for familial adenomatous polyposis. AB - AIMS: Restorative proctocolectomy with ileoanal anastomosis is one of the treatments of choice for patients suffering from familial adenomatous polyposis (FAP). However, any residual rectal mucosa left behind after mucosectomy is at risk for the development of neoplasia. CASE REPORT: A 31 year old male patient with FAP underwent restorative proctocolectomy with a pelvic ileal pouch-anal anastomosis. Seven years later he presented with right inguinal and perianal pain. A diagnosis of invasive columnar cuff carcinoma was made. DISCUSSION: Islets of columnar epithelium may be left behind after restorative proctocolectomy, exposing the patient to later malignant change. This risk must be emphasised and prevented by regular surveillance of the anastomosis. PMID- 11034597 TI - Cross-talk in cell death signaling PMID- 11034595 TI - Unsolved mysteries of intestinal M cells. PMID- 11034598 TI - Apoptosis control in syncytia induced by the HIV type 1-envelope glycoprotein complex: role of mitochondria and caspases. AB - Syncytia arising from the fusion of cells expressing a lymphotropic HIV type 1 encoded envelope glycoprotein complex (Env) with cells expressing the CD4/CXC chemokine receptor 4 complex spontaneously undergo cell death. Here we show that this process is accompanied by caspase activation and signs of mitochondrial membrane permeabilization (MMP), including the release of intermembrane proteins such as cytochrome c (Cyt-c) and apoptosis-inducing factor (AIF) from mitochondria. In Env-induced syncytia, caspase inhibition did not suppress AIF- and Cyt-c translocation, yet it prevented all signs of nuclear apoptosis. Translocation of Bax to mitochondria led to MMP, which was inhibited by microinjected Bcl-2 protein or bcl-2 transfection. Bcl-2 also prevented the subsequent nuclear chromatin condensation and DNA fragmentation. The release of AIF occurred before that of Cyt-c and before caspase activation. Microinjection of AIF into syncytia sufficed to trigger rapid, caspase-independent Cyt-c release. Neutralization of endogenous AIF by injection of an antibody prevented all signs of spontaneous apoptosis occurring in syncytia, including the Cyt-c release and nuclear apoptosis. In contrast, Cyt-c neutralization only prevented nuclear apoptosis, and did not affect AIF release. Our results establish that the following molecular sequence governs apoptosis of Env-induced syncytia: Bax mediated/Bcl-2-inhibited MMP --> AIF release --> Cyt-c release --> caspase activation --> nuclear apoptosis. PMID- 11034599 TI - Combined stimulation with the T helper cell type 2 cytokines interleukin (IL)-4 and IL-10 induces mouse mast cell apoptosis. AB - Mast cells are found in connective and mucosal tissues throughout the body. Their activation via immunoglobulin E (IgE)-antigen interactions is promoted by T helper cell type 2 (Th2) cytokines and leads to the sequelae of allergic disease. We now report a mechanism by which Th2 cytokines can regulate mast cell survival. Specifically, we find that interleukin (IL)-4 and IL-10 induce apoptosis in IL-3 dependent bone marrow-derived mast cells and peritoneal mast cells. This process required 6 d of costimulation with IL-3, IL-4, and IL-10, and expression of signal transducer and activator of transcription 6 (Stat6). Apoptosis was coupled with decreased expression of bcl-x(L) and bcl-2. While this process occurred independent of the Fas pathway, culture in IL-3+IL-4+IL-10 greatly sensitized mast cells to Fas-mediated death. Additionally, we found that IgE cross-linkage or stimulation with stem cell factor enhanced the apoptotic abilities of IL-4 and IL-10. Finally, IL-3-independent mastocytomas and mast cell lines were resistant to apoptosis induced by IL-3+IL-4+IL-10. These data offer evidence of Th2 cytokine-mediated homeostasis whereby these cytokines both elicit and limit allergic responses. Dysregulation of this pathway may play a role in allergic disease and mast cell tumor survival. PMID- 11034600 TI - T cells compete for access to antigen-bearing antigen-presenting cells. AB - These studies tested whether antigenic competition between T cells occurs. We generated CD8(+) T cell responses in H-2(b) mice against the dominant ovalbumin epitope SIINFEKL (ova8) and subdominant epitope KRVVFDKL, using either vaccinia virus expressing ovalbumin (VV-ova) or peptide-pulsed dendritic cells. CD8(+) T cell responses were visualized by major histocompatibility complex class I peptide tetrameric molecules. Transfer of transgenic T cells with high affinity for ova8 (OT1 T cells) completely inhibited the response of host antigen-specific T cells to either antigen, demonstrating that T cells can directly compete with each other for response to antigen. OT1 cells also inhibited CD8(+) T cell responses to an unrelated peptide, SIYRYGGL, providing it was presented on the same dendritic cells as ova8. These inhibitions were not due to a more rapid clearance of virus or antigen-presenting cells (APCs) by the OT1 cells. Rather, the inhibition was caused by competition for antigen and antigen-bearing cells, since it could be overcome by the injection of large numbers of antigen-pulsed dendritic cells. These results imply that common properties of T cell responses, such as epitope dominance and secondary response affinity maturation, are the result of competitive interactions between antigen-bearing APC and T cell subsets. PMID- 11034601 TI - Regulation of cytokine signaling by B cell antigen receptor and CD40-controlled expression of heparan sulfate proteoglycans. AB - Recently, biochemical, cell biological, and genetic studies have converged to reveal that integral membrane heparan sulfate proteoglycans (HSPGs) are critical regulators of growth and differentiation of epithelial and connective tissues. As a large number of cytokines involved in lymphoid tissue homeostasis or inflammation contain potential HS-binding domains, HSPGs presumably also play important roles in the regulation of the immune response. In this report, we explored the expression, regulation, and function of HSPGs on B lymphocytes. We demonstrate that activation of the B cell antigen receptor (BCR) and/or CD40 induces a strong transient expression of HSPGs on human tonsillar B cells. By means of these HSPGs, the activated B cells can bind hepatocyte growth factor (HGF), a cytokine that regulates integrin-mediated B cell adhesion and migration. This interaction with HGF is highly selective since the HSPGs did not bind the chemokine stromal cell-derived factor (SDF)-1 alpha, even though the affinities of HGF and SDF-1alpha for heparin are similar. On the activated B cells, we observed induction of a specific HSPG isoform of CD44 (CD44-HS), but not of other HSPGs such as syndecans or glypican-1. Interestingly, the expression of CD44-HS on B cells strongly promotes HGF-induced signaling, resulting in an HS-dependent enhanced phosphorylation of Met, the receptor tyrosine kinase for HGF, as well as downstream signaling molecules including Grb2-associated binder 1 (Gab1) and Akt/protein kinase B (PKB). Our results demonstrate that the BCR and CD40 control the expression of HSPGs, specifically CD44-HS. These HSPGs act as functional coreceptors that selectively promote cytokine signaling in B cells, suggesting a dynamic role for HSPGs in antigen-specific B cell differentiation. PMID- 11034603 TI - Requirements for bone marrow-derived antigen-presenting cells in priming cytotoxic T cell responses to intracellular pathogens. AB - Bone marrow (BM)-derived antigen-presenting cells (APCs) are potent stimulators of T cell immune responses. We investigated the requirements for antigen presentation by these cells in priming cytotoxic T lymphocyte (CTL) responses to intracellular bacterial and viral pathogens. [Parent-->F(1)] radiation BM chimeras were constructed using C57BL/6 donors and (C57BL/6 x BALB/c)F(1) recipients. Infection of chimeric mice with either Listeria monocytogenes or vaccinia virus expressing the nucleoprotein (NP) antigen from lymphocytic choriomeningitis virus (LCMV) primed H2-D(b)-restricted, but not H2-K(d) restricted CTL responses, demonstrating the requirement for BM-derived APCs for successful priming of CTL responses to these pathogens. Surprisingly, this did not hold true for chimeric mice infected with LCMV itself. LCMV-infected animals developed strong CTL responses specific for both H2-D(b)- and H2-L(d)-restricted NP epitopes. These findings indicate that in vivo priming of CTL responses to LCMV is remarkably insensitive to deficiencies in antigen presentation by professional BM-derived APCs. PMID- 11034602 TI - Transient inhibition of interleukin 4 signaling by T cell receptor ligation. AB - Interleukin (IL)-4 and IL-12 together with T cell receptor (TCR) engagement are crucial for the differentiation of CD4(+) T cells into T helper (Th)2 or Th1 cells, respectively. Although IL-4 receptors (IL-4Rs) but not IL-12Rs are expressed on naive CD4(+) T cells, IL-4 has no apparent advantage over IL-12 in driving naive T cell differentiation when the cells are primed with both IL-4 and IL-12 in vitro. It was found that IL-4-induced phosphorylation of Janus kinases 1 and 3, IL-4R alpha, signal transducer and activator of transcription 6, and insulin receptor substrate 2 was strikingly but transiently inhibited by TCR ligation both in conventional and TCR transgenic T cells. TCR engagement also blocked the expression of an IL-4-inducible gene. Signals induced by other cytokines, including IL-2, IL-6, and interferon alpha, but not by insulin-like growth factor 1, were also blocked by TCR engagement. The capacity of various inhibitors to reverse TCR-mediated inhibition of IL-4 signaling suggested that activation of the Ras-mitogen-activated protein kinase pathway and of the calcineurin pathway contribute to desensitizing IL-4R. IL-4 responsiveness returned at about the time ( approximately 12 h) that IL-12-mediated signaling was first observed. Thus, through different mechanisms, neither IL-4R nor IL-12R has any clear advantage in polarizing cells; rather, the availability of cytokine is probably the limiting factor in this process. PMID- 11034604 TI - Bone marrow-derived antigen-presenting cells are required for the generation of cytotoxic T lymphocyte responses to viruses and use transporter associated with antigen presentation (TAP)-dependent and -independent pathways of antigen presentation. AB - Bone marrow (BM)-derived professional antigen-presenting cells (pAPCs) are required for the generation of cytotoxic T lymphocyte (CTL) responses to vaccinia virus and poliovirus. Furthermore, these BM-derived pAPCs require a functional transporter associated with antigen presentation (TAP). In this report we analyze the requirements for BM-derived pAPCs and TAP in the initiation of CTL responses to lymphocytic choriomeningitis virus (LCMV) and influenza virus (Flu). Our results indicate a requirement for BM-derived pAPCs for the CTL responses to these viruses. However, we found that the generation of CTLs to one LCMV epitope (LCMV nucleoprotein 396-404) was dependent on BM-derived pAPCs but, surprisingly, TAP independent. The study of the CTL response to Flu confirmed the existence of this BM-derived pAPC-dependent/TAP-independent CTL response and indicated that the TAP-independent pathway is approximately 10-300-fold less efficient than the TAP-dependent pathway. PMID- 11034606 TI - FIST/HIPK3: a Fas/FADD-interacting serine/threonine kinase that induces FADD phosphorylation and inhibits fas-mediated Jun NH(2)-terminal kinase activation. AB - Fas is a cell surface death receptor that signals apoptosis. Several proteins have been identified that bind to the cytoplasmic death domain of Fas. Fas associated death domain (FADD), which couples Fas to procaspase-8, and Daxx, which couples Fas to the Jun NH(2)-terminal kinase pathway, bind independently to the Fas death domain. We have identified a 130-kD kinase designated Fas interacting serine/threonine kinase/homeodomain-interacting protein kinase (FIST/HIPK3) as a novel Fas-interacting protein. Binding to Fas is mediated by a conserved sequence in the COOH terminus of the protein. FIST/HIPK3 is widely expressed in mammalian tissues and is localized both in the nucleus and in the cytoplasm. In transfected cell lines, FIST/HIPK3 causes FADD phosphorylation, thereby promoting FIST/HIPK3-FADD-Fas interaction. Although Fas ligand-induced activation of Jun NH(2)-terminal kinase is impaired by overexpressed active FIST/HIPK3, cell death is not affected. These results suggest that Fas-associated FIST/HIPK3 modulates one of the two major signaling pathways of Fas. PMID- 11034605 TI - Immunoglobulin heavy chain variable region gene replacement As a mechanism for receptor revision in rheumatoid arthritis synovial tissue B lymphocytes. AB - Mature B cells can alter their antibody repertoires by several mechanisms, including immunoglobulin heavy chain variable region (V(H)) replacement. This process changes the antigen combining site by replacing a portion of the original V(H)/diversity/heavy chain joining region (V(H)DJ(H)) rearrangement with a corresponding portion of a new V(H) segment. This exchange can involve cryptic heptamer-like sequences embedded in the coding regions of V(H) genes. While studying the B lymphocytes that expand in the synovial tissues of patients with rheumatoid arthritis (RA), clones with V(H)DJ(H) variants that were apparently generated by V(H) replacement were identified with surprising frequency (approximately 8%). Examples of multiple independent V(H) replacement events occurring in distinct progeny clones were also identified. These secondary V(H) rearrangements were documented at both the cDNA and genomic DNA levels and involved several heptamer-like sequences at four distinct locations within V(H) (three sites in framework region 3 and one in complementarity determining region 2). The identification of blunt-ended double-stranded DNA breaks at the embedded heptamers and the demonstration of recombinase activating gene (RAG) expression suggested that these rearrangements could occur in the synovial tissues, presumably in pseudo-germinal centers, and that they could be mediated by RAG in a recognition signal sequence-specific manner. The presence of V(H) mutations in the clones that had undergone replacement indicated that these B cells were immunocompetent and could receive and respond to diversification signals. A relationship between these secondary V(H) gene rearrangements and the autoimmunity characteristic of RA should be considered. PMID- 11034607 TI - Nuclear factor kappa B is required for the development of marginal zone B lymphocytes. AB - Although immunoglobulin (Ig)M(hi)IgD(lo/-)CD21(hi) marginal zone B cells represent a significant proportion of naive peripheral splenic B lymphocytes, few of the genes that regulate their development have been identified. This subset of peripheral B cells fails to emerge in mice that lack nuclear factor (NF)-kappa Bp50. Less drastic reductions in marginal zone B cell numbers are also seen in the spleens of recombination activating gene (Rag)-2(-/-) mice reconstituted with NF-kappa Bp65(-/-) fetal liver cells and in c-Rel(-/-) mice. In contrast, steady state levels of IgD(hi) splenic follicular B cells are not significantly reduced in the absence of NF-kappa Bp50, NF-kappa Bp65, or c-Rel. Reconstitution of B cells in Rag-2(-/-) mice with a mixture of p50(-/-)/p65(-/-) fetal liver cells and Rag-2(-/-) bone marrow cells revealed that the generation of marginal zone B cells requires the expression of NF-kappa B in developing B cells, as opposed to supporting cells. PMID- 11034608 TI - Burkitt lymphoma in the mouse. AB - Chromosomal translocations juxtaposing the MYC protooncogene with regulatory sequences of immunoglobulin (Ig) H chain or kappa (Ig kappa) or lambda (Ig lambda) L chain genes and effecting deregulated expression of MYC are the hallmarks of human Burkitt lymphoma (BL). Here we report that lymphomas with striking similarities to BL develop in mice bearing a mutated human MYC gene controlled by a reconstructed Ig lambda locus encompassing all the elements required for establishment of locus control in vitro. Diffusely infiltrating lymphomas with a typical starry sky appearance occurred in multiple founders and an established line, indicating independence from positional effects. Monoclonal IgM(+)CD5(-)CD23(-) tumors developed from an initially polyclonal population of B cells. These results demonstrate that the phenotype of B lineage lymphomas induced by MYC dysregulation is highly dependent on cooperativity among the regulatory elements that govern expression of the protooncogene and provide a new system for studying the pathogenesis of BL. PMID- 11034609 TI - Genetic modulation of T cell receptor gene segment usage during somatic recombination. AB - Lymphocyte antigen receptors are not encoded by germline genes, but rather are produced by combinatorial joining between clusters of gene segments in somatic cells. Within a given cluster, gene segment usage during recombination is thought to be largely random, with biased representation in mature T lymphocytes resulting from protein-mediated selection of a subset of the total repertoire. Here we show that T cell receptor D beta and J beta gene segment usage is not random, but is patterned at the time of recombination. The hierarchy of gene segment usage is independent of gene segment proximity, but rather is influenced by the ability of the flanking recombination signal sequences (RSS) to bind the recombinase and/or to form a paired synaptic complex. Importantly, the relative frequency of gene segment usage established during recombination is very similar to that found after protein-mediated selection, suggesting that in addition to targeting recombinase activity, the RSS may have evolved to bias the naive repertoire in favor of useful gene products. PMID- 11034610 TI - Novel functional sets of lipid-derived mediators with antiinflammatory actions generated from omega-3 fatty acids via cyclooxygenase 2-nonsteroidal antiinflammatory drugs and transcellular processing. AB - Aspirin therapy inhibits prostaglandin biosynthesis without directly acting on lipoxygenases, yet via acetylation of cyclooxygenase 2 (COX-2) it leads to bioactive lipoxins (LXs) epimeric at carbon 15 (15-epi-LX, also termed aspirin triggered LX [ATL]). Here, we report that inflammatory exudates from mice treated with omega-3 polyunsaturated fatty acid and aspirin (ASA) generate a novel array of bioactive lipid signals. Human endothelial cells with upregulated COX-2 treated with ASA converted C20:5 omega-3 to 18R-hydroxyeicosapentaenoic acid (HEPE) and 15R-HEPE. Each was used by polymorphonuclear leukocytes to generate separate classes of novel trihydroxy-containing mediators, including 5-series 15R LX(5) and 5,12,18R-triHEPE. These new compounds proved to be potent inhibitors of human polymorphonuclear leukocyte transendothelial migration and infiltration in vivo (ATL analogue > 5,12,18R-triHEPE > 18R-HEPE). Acetaminophen and indomethacin also permitted 18R-HEPE and 15R-HEPE generation with recombinant COX-2 as well as omega-5 and omega-9 oxygenations of other fatty acids that act on hematologic cells. These findings establish new transcellular routes for producing arrays of bioactive lipid mediators via COX-2-nonsteroidal antiinflammatory drug-dependent oxygenations and cell-cell interactions that impact microinflammation. The generation of these and related compounds provides a novel mechanism(s) for the therapeutic benefits of omega-3 dietary supplementation, which may be important in inflammation, neoplasia, and vascular diseases. PMID- 11034611 TI - Visualization of Plasmodium falciparum-endothelium interactions in human microvasculature: mimicry of leukocyte recruitment. AB - Plasmodium falciparum-infected erythrocytes roll on and/or adhere to CD36, intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM) 1, and P-selectin under shear conditions in vitro. However, the lack of an adequate animal model has made it difficult to determine whether infected erythrocytes do indeed interact in vivo in microvessels. Therefore, we made use of an established model of human skin grafted onto severe combined immunodeficient (SCID) mice to directly visualize the human microvasculature by epifluorescence intravital microscopy. In all grafts examined, infected erythrocytes were observed to roll and/or adhere in not just postcapillary venules but also in arterioles. In contrast, occlusion of capillaries by infected erythrocytes was noted only in approximately half of the experiments. Administration of an anti-CD36 antibody resulted in a rapid reduction of rolling and adhesion. More importantly, already adherent cells quickly detached. The residual rolling after anti-CD36 treatment was largely inhibited by an anti-ICAM 1 antibody. Anti-ICAM-1 alone reduced the ability of infected erythrocytes to sustain rolling and subsequent adhesion. These findings provide conclusive evidence that infected erythrocytes interact within the human microvasculature in vivo by a multistep adhesive cascade that mimics the process of leukocyte recruitment. PMID- 11034612 TI - Cross-talk in cell death signaling. PMID- 11034613 TI - Keeping up with bicarbonate. PMID- 11034615 TI - Urea transport by cotransporters. AB - The rabbit Na+-glucose cotransporter (rbSGLT1) was expressed in Xenopus laevis oocytes and urea transport in rbSGLT1 and non-injected (control) oocytes was studied using [14C]urea as a tracer. The level of rbSGLT1 expression in these batches of oocytes was monitored by measuring the uptake of alpha-methyl-D [14C]glucopyranoside ([14C]alphaMDG). In rbSGLT1-expressing oocytes, there was a 4-fold increase in urea transport in the absence of sugar relative to that in control oocytes. Urea uptake was not Na+ dependent and was linear with both time of incubation (5-120 min) and increasing urea concentration (50 microM to 100 mM) in the bathing medium. rbSGLT1 urea uptake was blocked by the rbSGLT1-specific inhibitor phlorizin (Ki 1 microM) in 100 mM NaCl buffer, but was not affected in 100 mM choline chloride buffer. Phloretin inhibited rbSGLT1 urea uptake with a low affinity (Ki > 1 mM) in the presence and absence of Na+. The uptake of 55 m[mu]M urea through rbSGLT1 was not blocked by 100 mM urea analogues including thiourea, 1,3-dimethyl urea, 1,1-dimethyl urea and acetamide. The activation energies (Ea) of urea transport for control and rbSGLT1-expressing oocytes were 14+/-3 and 6+/-1 kcal mol(-1), respectively. The low Ea for urea transport through rbSGLT1 is comparable to the Ea of passive water transport through rbSGLT1. Urea transport through rbSGLT1 was further increased when the cotransporter was activated by the addition of sugar to the external medium. The rate of sugar-dependent urea uptake was directly proportional to the rate of Na+ glucose-H2O cotransport such that the amount of urea transport was approximately proportional to the molar concentration ratio of urea to H2O (55 microM/55 M). The low affinity Na+-glucose (pSGLT3), the Na+-iodide (rNIS) and the Na+-(Cl-) GABA (hGAT1) cotransporters expressed in oocytes demonstrated similar urea transport properties. These observations suggest that cotransporters behave as urea channels in the absence of substrates. Furthermore, under substrate transporting conditions, the same cotransporters serve as urea cotransporters. This could account for urea transport in cells that appear not to have urea uniporters or channels. PMID- 11034614 TI - Molecular determinants of inactivation in voltage-gated Ca2+ channels. AB - Evolution has created a large family of different classes of voltage-gated Ca2+ channels and a variety of additional splice variants with different inactivation properties. Inactivation controls the amount of Ca2+ entry during an action potential and is, therefore, believed to play an important role in tissue specific Ca2+ signalling. Furthermore, mutations in a neuronal Ca2+ channel (Ca(v)2.1) that are associated with the aetiology of neurological disorders such as familial hemiplegic migraine and ataxia cause significant changes in the process of channel inactivation. Ca2+ channels of a given subtype may inactivate by three different conformational changes: a fast and a slow voltage-dependent inactivation process and in some channel types by an additional Ca2+-dependent inactivation mechanism. Inactivation kinetics of Ca2+ channels are determined by the intrinsic properties of their pore-forming alpha1-subunits and by interactions with other channel subunits. This review focuses on structural determinants of Ca2+ channel inactivation in different parts of Ca2+ channel alpha1-subunits, including pore-forming transmembrane segments and loops, intracellular domain linkers and the carboxyl terminus. Inactivation is also affected by the interaction of the alpha1-subunits with auxiliary beta-subunits and intracellular regulator proteins. The evidence shows that pore-forming S6 segments and conformational changes in extra- (pore loop) and intracellular linkers connected to pore-forming segments may play a principal role in the modulation of Ca2+ channel inactivation. Structural concepts of Ca2+ channel inactivation are discussed. PMID- 11034616 TI - The non-hydrolytic pathway of cystic fibrosis transmembrane conductance regulator ion channel gating. AB - It has been suggested that the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel may utilize a novel gating mechanism in which open and closed states are not in thermodynamic equilibrium. This suggestion is based on the assumption that energy of ATP hydrolysis drives the gating cycle. We demonstrate that CFTR channel gating occurs in the absence of ATP hydrolysis and hence does not depend on an input of free energy from this source. The binding of ATP or structurally related analogues that are poorly or non-hydrolysable is sufficient to induce opening. Closing occurs on dissociation of these ligands or the hydrolysis products of those that can be cleaved. Not only can channel opening occur without ATP hydrolysis but the temperature dependence of the open probability (Po.) is reversed, i.e. Po. increases as temperature is lowered whereas under hydrolytic conditions, Po. increases as temperature is elevated. This indicates that there are different rate-limiting steps in the alternate gating pathways (hydrolytic and non-hydrolytic). These observations demonstrate that phosphorylated CFTR behaves as a conventional ligand-gated channel employing cytoplasmic ATP as a readily available cytoplasmic ligand; under physiological conditions ligand hydrolysis provides efficient reversibility of channel opening. PMID- 11034617 TI - A single residue contributes to the difference between Kir4.1 and Kir1.1 channels in pH sensitivity, rectification and single channel conductance. AB - Kir1.1 and Kir4.1 channels may be involved in the maintenance of pH and K+ homeostasis in renal epithelial cells and CO2 chemoreception in brainstem neurons. To understand the molecular determinants for their characteristic differences, the structure-function relationship was studied using site-directed mutagenesis. According to previous studies, Glu158 in Kir4.1 is likely to be the major rectification controller. This was confirmed in both Kir1.1 and Kir4.1. Mutation of Gly210, the second potential rectification controller, to glutamate did not show any additional effect on the inward rectification. More interestingly, we found that Glu158 in Kir4.1 was also an important residue contributing to single channel conductance and pH sensitivity. The E158N Kir4.1 mutant had a unitary conductance of 35 pS and a midpoint pH for channel inhibition (pKa) value of 6.72, both of which were almost identical to those of the wild-type (WT) Kir1.1. Flickering channel activity was clearly seen in the E158N mutant at positive membrane potentials, which is typical in the WT Kir1.1 but absent in the WT Kir4.1. Reverse mutation in Kir1.1 (N171E) reduced the unitary conductance to 27 pS (23 pS in WT Kir4.1). However, the pH sensitivity of this mutant did not show a marked difference from the WT Kir1.1. Therefore, it is possible that a residue(s) in addition to Asn171 is also involved. Thus we studied several other residues in both M2 and H5 regions. We found that joint mutations of Val140 and Asn171 to residues seen in Kir4.1 greatly reduced the pH sensitivity (pKa 6. 08). The V140T mutation in Kir1.1 led to a unitary conductance of approximately 70 pS, and the G210E mutation in Kir4.1 caused a decrease in pH sensitivity of 0.4 pH units. These results indicate that the pore forming sequences are targets for modulations of multiple channel-biophysical properties and demonstrate a site contributing to rectification, unitary conductance and proton sensitivity in these Kir channels. PMID- 11034618 TI - Muscarinic inhibitory and stimulatory regulation of the L-type Ca2+ current is not altered in cardiac ventricular myocytes from mice lacking endothelial nitric oxide synthase. AB - Using conventional and perforated patch-clamp techniques, the inhibitory and stimulatory effects of acetylcholine (ACh) on beta-adrenergic regulation of the L type Ca2+ current (ICa) were studied in ventricular myocytes from wild-type mice (WT) and from mice lacking endothelial nitric oxide synthase (eNOS or NOS3; NOS3 KO mice). To validate the direct comparison of ACh effects on beta-adrenergic responses, the sensitivity of ICa to the beta-adrenergic agonist isoprenaline (Iso) was studied in both WT and NOS3-KO mouse myocytes. ICa sensitivity to Iso was not found to be significantly different in WT and NOS3-KO myocytes: Iso increased ICa with an EC50 of 4.9 and 3.7 nM in WT and NOS3-KO myocytes, respectively. ACh-induced inhibition of ICa did not significantly differ in ventricular myocytes from WT and NOS3-KO mice. ACh (10 microM) inhibited the stimulatory effect of 3 nM Iso by 39 and 35% in WT and NOS3-KO myocytes, respectively. Exposure to and subsequent washout of ACh in the continuous presence of submaximally stimulating concentrations of Iso (1-3 nM) resulted in a transient rebound stimulation of ICa in both WT and NOS3-KO mouse myocytes. The magnitude of the stimulatory effect of ACh did not significantly differ in WT and NOS3-KO mice. These results indicate that nitric oxide (NO) generated by NOS3 does not significantly affect the beta-adrenergic responsiveness of ICa. The results also confirm previous work indicating that NO generated by NOS3 is not obligatory for muscarinic inhibition of the beta-adrenergically regulated ICa in ventricular myocytes. Finally these results demonstrate for the first time that NO generated by NOS3 is not involved in muscarinic rebound stimulation of ICa in ventricular myocytes.